PMID- 12751494 TI - Clinical trials face heightened scrutiny as science and commerce appear to merge. AB - Clinical trials involving human subjects face increased scrutiny and regulation by the federal government that may lead to costly changes to the process. Prompted by a highly publicized death in a gene therapy trial, federal regulators are examining almost all aspects of clinical trial conduct, from how patients are recruited and how they give informed consent, to the relationship between the researchers and the company sponsoring the research. In addition to the heightened regulation, the government has announced that it will seek the power to impose civil money penalties on researchers who do not observe applicable patient protection regulations. In an effort to include more senior citizens in clinical trials, the government for the first time will cover certain costs of Medicare beneficiaries who participate in clinical trials. The risk of increased government involvement includes criminal prosecution of physicians for failing to follow the rules, as an innovative case in California demonstrates. PMID- 12751495 TI - Mapping of human genome raises serious issues for individual vs. corporate rights. AB - Unrestricted corporate control of genetic technologies opens a pandora's box for the balance of power between individuals and corporations. This column provides an overview of the issues raised by commercialization of the human genome from a citizen advocate's perspective. PMID- 12751496 TI - An analysis of the proposal for population based screening for hereditary hemochromatosis in Massachusetts. AB - The Massachusetts Senate is considering a bill which would require a screening test for iron overload diseases when such a test is medically necessary. The legislation is also authorized to establish, promote, and administer a statewide early detection program for the detection of hereditary hemochromatosis. Despite the challenges to a successful screening program which have been documented in the medical literature and the potential for genetic discrimination, the public health benefits justify this legislation as long as the necessary modifications and precautions are established. PMID- 12751497 TI - Hygiene, health, and bodily knowledge, 1880-1940: a New Zealand case study. AB - One of the key aims of the Dunedin Southern Suburbs project has been to examine the factors that structured opportunity in a period of rapid modernization in New Zealand, 1880-1940. This article argues that health was a key determinant of opportunity and examines how people thought about their bodies and sought to maintain health. It argues that older, humoral theories about health were maintained alongside new understandings wrought by the germ theory and that both placed a high value on regimen. Mothers were regarded as the household experts on health but lacked a vocabulary to communicate with their daughters about menstruation. While bodily health was an ideal to be striven for, there was little communication about bodily pleasure, a situation subject to change with the adoption of family limitation. PMID- 12751498 TI - Pharmaceutical advertising as a consumer empowerment device. AB - Pharmaceutical companies have greatly increased their level of "direct-to consumer" (DTC) advertising in recent years. For 1998, estimates are that over $1.1 billion was spent on this form of advertising, increased from $850 million in 1997 and $600 million in 1996. In 1998, 84 separate drugs were advertised to consumers. The impetus was a decision in August of 1997 by the Food and Drug Administration to reduce the restrictions on DTC advertising on television. As a result, such ads have become very common on TV, and 32 products were advertised on TV in 1998. Pharmaceutical companies advertise because they think that advertising will make money for them. But how will this make money? It will make money by providing consumers with the information they need to make proper decisions about medication. That is, DTC advertising is profitable exactly because it empowers consumers and enables them to purchase useful drugs. The goals of advertising companies and consumers are both for consumers to have information about the most beneficial drug for particular conditions, and so advertising is beneficial both to manufacturers and to consumers. This article describes emerging trends in DTC within the context of the life sciences sector. PMID- 12751499 TI - [A battle of th health clubs in the Netherlands around 1840]. AB - The discovery of a national inquiry into health funds in the 1840s gives cause to reconsider the traditional view on this subject. After a prosperous period under the guild regime commercial interests penetrated the health market. Its directors supposedly enriched themselves at the expense of both the insured and the professionals (general practitioners (gp's) and chemists). As things grew worse the government intervened and ordered an inquiry by a Select Committee. In reality things were slightly different. The debate on heath insurance was part and parcel of the pursuit of the medical profession to improve its standing. An alarming report on health funds by the Amsterdam medical commission spurred the Health Department to action. The national inquiry did not in fact corroborate the earlier report. In most parts of the Netherlands health insurance did not exist. In only two provinces - North and South Holland - taking out health insurance was an option and even there it was to a large extent in the cities. The three big cities - Amsterdam, Rotterdam and The Hague - accounted for 51.8 percent of all funds and as much as 72.2 percent of all persons insured. Nearly all complaints in the 1840s originated from gp's in Amsterdam. Only The Hague had the same experience i.e. infringements on private practice, enrichment by the fund governors etc. The complaints were not unlike those of the English club doctors at the end of the century. In both cities commercial health funds were important, while in Rotterdam gp's had very often been founding fathers. The author has tried to put the complaints into a wider perspective by comparing commercial funds with local clubs' and gp's own fund, the AZA, founded in 1847 to combat the disputed trend. Finally commercial funds have been compared with medical relief (number of patients and paid fees). The complaints about remuneration seem exaggerated as even AZA could not afford to pay higher fees. Commercialisation had only just started, but the new funds were more successful than the older clubs. AZA, however, proved to be even more successful. In close co-operation with poor law authorities a demarcation was introduced between medical relief and the working poor who had to take out a health insurance. AZA excluded the well-to do from participating. Amsterdam developed a brand new health insurance policy in response to the threat of commercial funds. In the early twentieth century the story was repeated on a national level in response to proposals for compulsory sickness insurance. PMID- 12751500 TI - The regulation of home diagnostic tests for genetic disorders: can the FDA deny a premarket application on the basis of the device's social impacts? AB - This article discusses the Food and Drug Administration (FDA) authority to regulate home use diagnostics for genetic disorders based on the social effects of the devices in the premarket approval process (PMA). It begins with a discussion of the potential social impacts of home use genetic diagnostics, focusing particularly on the psychological effects of the devices. The article then discusses the FDA's past experience in regulating home use diagnostics for HIV and for drugs of abuse testing under its PMA authority. In approving PMA applications for home tests for HIV and drugs of abuse, the FDA has considered various social effects of the devices and tailored its approval to these considerations. However, the agency did not deny approval because of social considerations. The author argues that the FDA's past experience in approving PMA applications indicates both a willingness to consider social considerations in the PMA process and that the agency may deny a PMA application based on social considerations in appropriate circumstances. The author also argues that the FDA may deny application if the social impacts and other health risks of the device greatly outweigh the therapeutic benefits of the device. For example, in applications involving late onset genetic disorders for which no therapeutic options currently exist, the social risks of the device may be found to outweigh the benefits to health and therefore the FDA may deny a PMA application in these situations. PMID- 12751501 TI - [Dr. Jakob Lutz, life and work]. AB - The life and work of the Zurich child and youth psychiatrist, Prof. Dr. med. J. Lutz, (25.1.1903 - 28.6.1998) is the subject of this report. It documents the outstanding achievements of Jakob Lutz as a doctor and scientist and tries to give a true portrayal of his impressive personality. A biographical part is dedicated to the personal and professional development of the son of a family of train drivers from Appenzell, who, after studying medicine in Zurich, became the founder of the child and youth psychiatric service in the Canton of Zurich. A detailed report is given on the formation of the child an youth psychiatric service in the Canton of Zurich over almost 40 years. It begins in 1929, when med. prakt. Jakob Lutz took over management of the Stephansburg, the first observation unit for child psychiatry in Switzerland, and ends in 1968, when he retired, after becoming Director of the service an both extraordinary professor and honorary professor for child an youth psychiatry. Discussions on different forms of therapy in child an youth psychiatry include ideas on the therapeutic activity of Jakob Lutz himself and his demands on the personality and training of therapist in child and youth psychiatry. The chapter on child schizophrenia, autism and mentally handicapped children deals with special patient groups. All the chapters report on the specific medical scientific work of Jakob Lutz, always supplemented with quotations from his personal statements. Finally, there is also a brief assessment of the successor in the service, Prof. Dr. med. J. Corboz, the first ordinary professor for child and youth psychiatry at the University of Zurich. A list of Jakob Lutz's publications, a list of the lecture titles, and time charts supplement the report. PMID- 12751502 TI - John Trevisa's translation of Bartholomaeus Anglicus' De proprietatibus rerum. PMID- 12751503 TI - Nicholas de Cusa (1401-1464): a description of fine grained and polished wooden spoons employed as mirrors; concave, convex, flat and cylindrical (1450 AD). PMID- 12751504 TI - Medicaid prescription drug coverage: state efforts to control costs. AB - This paper provides a brief summary of the Medicaid prescription drug benefit. It explains the mechanisms being used by states to control their prescription drug spending within the Medicaid program. The paper also highlights some of the concerns that have been expressed with these mechanisms and the litigation that has been initiated in several states as a result of these efforts. It takes a closer look at three states with cost-containment strategies that have been the focus of increased scrutiny. PMID- 12751505 TI - Survey of change in practice following simulation-based training in crisis management. AB - We investigated the long-term effects on clinical practice of a simulation-based course in anaesthesia crisis management. A questionnaire was posted to all anaesthetists who had attended a course in the preceding year. The response rate was 69% (66/96). The crisis management course was valued highly by respondents, who perceive a change in practice as a result of the training. This change in practice was not limited to the specific clinical events simulated in the course, but applied to a wide range of events and to routine practice. The high rate of subsequent critical events reported in the survey supports the need for training in this area. This survey suggests that simulation-based training in crisis management is an effective form of continuing medical education for anaesthetists. PMID- 12751506 TI - Analgesia during radial artery cannulation: comparison of the effects of lidocaine applied by local injection or iontophoresis. AB - We compared the effect of lidocaine injection with lidocaine iontophoresis for pain relief during radial artery cannulation prior to induction of anaesthesia. Patients were allocated randomly to one of two groups. Group 1 (n = 15) received iontophoresis for 10 min prior to cannulation, using 4% lidocaine 4 ml. Group 2 (n = 15) received local infiltration of 1% lidocaine 1 ml using a 25G needle. Pain scores were recorded immediately after cannulation using a 10-cm visual analogue scale (VAS). There was no difference in mean (SD) pain scores [group 1: 2.2 (1.5) cm; group 2: 2.3 (2.7) cm; 95% CI of difference -1.8 to 1.5 cm]. Lidocaine delivered by iontophoresis is an effective and painless method of providing analgesia for radial artery cannulation. PMID- 12751508 TI - Current world literature. PMID- 12751507 TI - Survey of use of end-tidal carbon dioxide for confirming tracheal tube placement in intensive care units in the UK. AB - The use of end-tidal carbon dioxide monitoring to confirm the correct placement of a tracheal tube immediately after intubation is mandatory in the operating theatre. Tracheal intubation in critically ill patients can be challenging. Quick and accurate confirmation of tracheal tube placement is essential to minimise complications. This survey explored the use of end-tidal carbon dioxide monitoring to confirm tracheal tube placement in intensive care units in the UK. Questionnaires were sent to either the lead clinician or clinical director of randomly selected general adult intensive care units. One hundred and twenty seven replies were received from the 215 questionnaires sent (response rate 59%). Twenty per cent of the units did not have an end-tidal carbon dioxide monitor, 20% had one end-tidal carbon dioxide monitor per bed and 60% had one end-tidal carbon dioxide monitor between several beds. Only 50% of the units having an end tidal carbon dioxide monitor use it to confirm correct tracheal tube placement. Of these 50%, only about a third use it for every intubation. Seventy-two per cent of respondents felt that end-tidal carbon dioxide is well suited to confirm correct placement of tracheal tube in critically ill patients, but 50% did not think that confirmation using end-tidal carbon dioxide should be mandatory for intubations outside the operating theatre. Half of the units not having end-tidal a carbon dioxide monitor cited lack of resources as a reason. In summary, although four in every five intensive care units surveyed have end-tidal carbon dioxide monitors, only a small proportion use them to confirm correct placement of tracheal tube after intubation. PMID- 12751509 TI - Group B beta haemolytic streptococcal (GBS) infection in the fetus is not always an ascending infection. PMID- 12751510 TI - Insertion of an inferior venocaval filter in a pregnant woman at risk for pulmonary embolism-- a challenging management. PMID- 12751511 TI - Addison's disease, with successful pregnancy outcome. PMID- 12751512 TI - The use of total parenteral nutrition in protracted hyperemesis gravidarum. PMID- 12751513 TI - Raising awareness of postpartum seizures. PMID- 12751514 TI - Maternal hepatic dysfunction in the third trimester of pregnancy in an infant with fatty oxidation defect. PMID- 12751515 TI - Phaeochromocytoma-- an unusual cause of fitting in pregnancy. PMID- 12751516 TI - Intractable hiccoughs in pregnancy. PMID- 12751517 TI - Maternal collapse and coagulopathy following sexual intercourse in pregnancy. PMID- 12751518 TI - New angles and cavities. PMID- 12751519 TI - Terminating e-trafficking. PMID- 12751520 TI - Spontaneous resolution of HELLP syndrome in a primigravida presenting at 31 weeks' gestation. PMID- 12751521 TI - Liquid array single-handedly detects bounty of BW agents. PMID- 12751523 TI - On-line particle monitoring. PMID- 12751522 TI - Two cases of early-onset obstetric cholestasis. PMID- 12751524 TI - Low-tech chemical detector badge. PMID- 12751525 TI - Silent, massive haemoperitoneum during labour secondary to a spontaneous rupture of a utero-ovarian vessel. PMID- 12751526 TI - Responsive drug delivery systems. PMID- 12751527 TI - Sole bladder metastasis from breast cancer. PMID- 12751528 TI - Emily Bronte's homesickness. PMID- 12751529 TI - FTIR spectra database of inorganic art materials. PMID- 12751530 TI - Epidermoid cyst of the ovary. PMID- 12751531 TI - Bulwer's misanthropes and the limits of Victorian sympathy. PMID- 12751532 TI - Shedding light on NSOM. PMID- 12751533 TI - Tubo-ovarian abscess resembling ovarian malignancy. PMID- 12751534 TI - Reducing default from colposcopy clinics. PMID- 12751535 TI - Ethics self-assessment. PMID- 12751536 TI - Advantages of having a practice Web site. PMID- 12751537 TI - Black AIDS campaign launched in Bay area. PMID- 12751538 TI - Interferon may help early HIV infection. PMID- 12751540 TI - TV getting better with safer sex. PMID- 12751539 TI - Resistant HIV in breast milk. PMID- 12751541 TI - New rapid HIV test. PMID- 12751542 TI - Sex workers my spread HIV here. PMID- 12751543 TI - New face of AIDS in San Francisco. PMID- 12751544 TI - Staph skin infection outbreak in Los Angeles. PMID- 12751545 TI - Toxicity of anaesthetics. AB - Unwanted side-effects of anaesthetic drugs that occur during anaesthesia or during the post-operative recovery period are what concern anaesthesiologists and anaesthetists. Occupational risks are of concern to all health-care professionals who administer anaesthetics or who are incidentally exposed to anaesthetic gases. After regulatory requirements for marketing drugs are met, the qualitative and quantitative nature of side-effects of the drugs in the target population and the risk of incidental exposure of health-care professionals are generally well defined. However, sometimes unwanted side-effects are first detected after post approval marketing. Numerous scientific disciplines with specialized terminology contribute to the body of knowledge about anaesthetic toxicity. Scientific inquiry spanning a range of disciplines from molecular biology to global ecology provides information essential for predicting, assessing, avoiding and treating the untoward effects of anaesthetics. Contemporary concerns with respect to side effects of anaesthetic drugs include delayed recovery of cognitive function, addiction and tolerance, local anaesthetic cardiotoxicity and tissue toxicity, relative toxicity of enantiomeric forms of drugs, and the role of biotransformation in unwanted responses to anaesthetic drug administration. PMID- 12751546 TI - What is toxicology and how does toxicity occur? AB - Toxicology has matured since it was defined as the 'science of poisons'. Modern toxicology is no longer anthropocentric but takes on different views at various biological systems, including ecosystems. Each will interact specifically when exposed to defined chemical agents, including drugs. Adverse effects during drug therapy or after (accidental) poisoning are the result of some negative interactions between the agent and the exposed biological system. Toxicity is no longer a specific property of drugs and chemicals but an operative term to describe the adverse outcome of a specific drugs-host interaction. Newer developments in toxicology have focused on the host. Toxicogenetics continues to provide answers to variations of host response to xenobiotics, including drugs. Clinically relevant genetic polymorphisms and gene defects have been detected, and their number is rapidly growing. The key to understanding is in the host proteins that interact with the drug and mediate the cellular response. Hence, the proteom, i.e. the complete set of proteins of a cell, an individual or a species, determines how an exposed biological system may interact with the manifold of different xenobiotics. Structure-activity studies try to find out useful predictive parameters for risk and toxicity assessment. PMID- 12751547 TI - Halogenated inhalational anaesthetics. AB - The halogenated inhalational anaesthetics halothane, enflurane, isoflurane and desflurane can produce metabolic hepatocellular injury in humans to a variable extent. During metabolism of these anaesthetics, tissue acetylation occurs due to the formation of reactive intermediates. Proteins modified by acetylation may constitute neo-antigens with a potential for triggering an antibody-mediated immune response. The likelihood of suffering post-operative immune hepatitis depends on the amount of the anaesthetic metabolized and is thereby considerably less with enflurane, isoflurane or desflurane compared with halothane. Plasma inorganic fluoride concentrations are regularly increased after sevoflurane. Elevated inorganic fluoride concentrations have been associated with nephrotoxicity following methoxyflurane anaesthesia but not after sevoflurane. Another source of concern is the products of degradation from reactions with carbon dioxide absorbents. Most important is compound A, which has been shown to exhibit nephrotoxicity in rodents. However, no significant changes in renal function parameters have been reported in surgical patients. PMID- 12751548 TI - Toxicity of nitrous oxide. AB - Nitrous oxide interacts with vitamin B12 resulting in selective inhibition of methionine synthase, a key enzyme in methionine and folate metabolism. Thus, nitrous oxide may alter one-carbon and methyl-group transfer most important for DNA, purine and thymidylate synthesis. Long-term exposure to high concentrations of nitrous oxide may cause megaloblastic bone-marrow depression and neurological symptoms. Exposure to higher doses for less than 6 hours, as in clinical anaesthesia, are considered harmless. Recent studies seem to suggest a correlation between nitrous oxide anaesthesia and hyperhomocysteinaemia which is accepted to be an independent risk factor for coronary artery disease. As for today, available data do not support the notion that exposure to trace amounts of nitrous oxide is associated with impaired fertility or an increased risk of developing cancer. Emission of nitrous oxide from medical use is estimated to contribute less than 0.05% to total annual greenhouse gas emission. PMID- 12751549 TI - Interaction of inhalational anaesthetics with CO2 absorbents. AB - We review the currently available carbon dioxide absorbents: sodium hydroxide lime (=soda lime), barium hydroxide lime, potassium-hydroxide-free soda lime, calcium hydroxide lime and non-caustic lime. In general, all of these carbon dioxide absorbents are liable to react with inhalational anaesthetics. However, there is a decreasing reactivity of the different absorbents with inhalational anaesthetics: barium hydroxide lime >> soda lime > potassium-hydroxide-free soda lime > calcium hydroxide lime and non-caustic lime. Gaseous compounds generated by the reaction of the anaesthetics with desiccated absorbents are those that threaten patients. All measures are comprehensively described to--as far as possible--prevent any accidental drying out of the absorbent. Whether or not compound A, a gaseous compound formed by the reaction of sevoflurane with normally hydrated absorbents, is still a matter of concern is discussed. Even after very high loading with this compound, during long-lasting low-flow sevoflurane anaesthesias, no clinical or laboratory signs of renal impairment were observed in any of the surgical patients. Finally, guidelines for the judicious use of different absorbents are given. PMID- 12751550 TI - Toxicity of intravenous anaesthetics. AB - Intravenous anaesthetic agents are generally remarkably safe. However, it is clear that propofol infusion syndrome is a real, albeit rare, entity. This often lethal syndrome of metabolic acidosis, acute cardiomyopathy and skeletal myopathy is strongly associated with infusions of propofol at rates of 5 mg/kg/hour and greater for more than 48 hours. There is evidence to support the hypothesis that the syndrome is caused by the failure of free fatty acid metabolism due to inhibition of free fatty acid entry into the mitochondria and also specific sites in the mitochondrial respiratory chain. The syndrome therefore mimics the mitochondrial myopathies. Midazolam causes seizure-like activity in very-low birthweight premature infants requiring the drug prior to tracheal intubation or during prolonged positive pressure ventilation. This can be successfully reversed with the specific benzodiazepine antagonist flumazenil. Midazolam can also cause paradoxical reactions, including increased agitation, poor co-operation and aggressive or violent behaviour, which has been successfully managed with flumazenil. PMID- 12751551 TI - Opioid and non-opioid analgesics. AB - Opioids are the most potent analgesics. Toxicity results either from effects mediated by variation in affinity and intrinsic efficacy at specific opioid receptors or, rarely, from a direct toxic effect of the drugs. For some adverse effects, opioids exhibit a 'dual pharmacology' whereby these effects are usually observed only in pain-free individuals, and are not seen in patients in pain. Paracetamol, although generally very safe in therapeutic doses, displays potentially fatal toxicity in overdose requiring specific treatment. Non steroidal anti-inflammatory drugs (NSAIDs) are known to act by inhibiting COX-1 and COX-2 isoenzymes to various degrees. Toxicity arises primarily from undesired inhibition at these enzyme sites. Knowledge of the mechanism of action of these drugs is fundamental to the understanding of their potential for toxicity, the details of which are still emerging. PMID- 12751552 TI - Toxicity of local anaesthetics. AB - The complications of failure, neural injury and local anaesthetic toxicity are common to all regional anaesthetic techniques, and individual techniques are associated with specific complications. All potential candidates for regional anaesthesia should be thoroughly evaluated and informed of potential complications. Central neural blockades still account for more than 70% of regional anaesthesia procedures. Permanent neurological injury is 0.02-0.07%. Pain on injection and paraesthesias while performing regional anaesthesia are danger signals of potential injury and must not be ignored. The incidence of systemic toxicity to local anaesthetics has significantly decreased in the past 30 years, from 0.2 to 0.01%. Peripheral nerve blocks are associated with the highest incidence of systemic toxicity (7.5 per 10,000) and the lowest incidence of serious neural injury (1.9 per 10,000). PMID- 12751553 TI - Muscle relaxants. AB - Studies on the toxic effects of muscle relaxants are difficult to design because of the need for mechanical ventilation and, consequently, concomitant administration of anaesthetic drugs which may influence the results. The following overview shows that muscle relaxants are weak toxic agents with regard to their teratogenicity, carcinogenicity and cytotoxic effects (including tissue- and organ-damaging effects). Moreover, this chapter presents other side-effects of muscle relaxants under the broad heading of toxicity: the succinylcholine triggered cytotoxic effects on skeletal muscle cells with different aetiology, for example, or persistent muscle weakness after long-term administration of non depolarizing muscle relaxants. Receptor stimulation in the central nervous system may cause acute excitement and seizures. Muscle relaxants and their metabolites may interact with muscarinic and nicotinic receptors in other organs and the ganglionic system, for example in the cardiovascular system. Direct stimulation of mast cells, with consequent release of histamine, after administration of muscle relaxants may clinically impose as toxic reactions. PMID- 12751554 TI - Occupational hazards of inhalational anaesthetics. AB - Occupational exposure to inhalational anaesthetics has often been associated with health hazards and reproductive toxicity, but the available evidence is weak and comes mostly from epidemiological studies that have been criticized. Studies based on registered data generally showed no association between occupational exposure to inhalational anaesthetics and reproductive effects. Animal studies also showed a lack of carcinogenicity, organ toxicity and reproductive effects with trace concentrations, as observed in operating rooms. The exception may be nitrous oxide, which in some, but not all, studies showed teratogenicity in rats chronically exposed to concentrations of 1000 p.p.m. and higher, such as may occur in unscavenged operating rooms lacking a mechanical ventilation system. Occupational exposure has also been associated with impairment of psychological functions, but these effects do not occur with trace concentrations. All in all, the scientific evidence for hazards is weak. Nonetheless, it is good practice to limit levels of exposure. PMID- 12751555 TI - Inverted papilloma of the nose and paranasal sinuses: a fifteen-year review. AB - Inverted papilloma is a relatively rare epithelial neoplasm of the nose and paranasal sinuses accounting for 0.5-4% of all primary tumours of the nose. This is a retrospective study of fifteen patients with histologically confirmed I.P. of the nose and paranasal sinuses seen from 1986 to 2000 at the Department of Otorhinolaryngology, University College Hospital, Ibadan. The patients ages range from 14-65 years, with a median age of 39 years. There were twelve males and three females. Five patients had advanced lesions of which four were bilateral. There was associated synchronous and metachronous squamous cell carcinoma (SCC) in two patients respectively. The surgery offered was based on the clinical and radiological evaluations of these patients. Nine patients had conservative surgery (intranasal clearance/antrostomy and external fronto-ethmoidectomy) while the remaining six patients had radical surgery (lateral rhinotomy/medial maxillectomy) at various times. The two patients with SCC had adjunct radiotherapy/chemotherapy in addition. Recurrence was observed in those that had conservative surgery only. We therefore advocate radical surgery in our environment as the treatment of choice for effective limitation of recurrence. PMID- 12751556 TI - Effects of some risk factors and immunodeficiencies on the periodontium--a review. AB - The possible role systemic factors might play in initiating or modifying the progress of periodontal disease has been a controversial issue for some time. Gingivitis is initiated by microbial plaque deposits on the dento-gingival interface but progression to periodontitis is modified by several environmental, behavioural, biological and health care variables. The importance of the immune system in modifying the host response to plaque is well established and as such, the immune system is a risk factor for human and animal periodontal disease. This paper reviews the modifying risk factors for periodontal disease and examines the periodontal manifestations of subjects with primary and acquired immuno deficiencies. PMID- 12751557 TI - Problems of widowhood: a study of widows in a tertiary institution in Ibadan, south west Nigeria. AB - This study is part of a larger multi-centre survey on widowhood in Nigeria. Information was gathered using a structured self-administered questionnaire from 42 widows who are working at the University College Hospital and the College of Medicine, in Ibadan, capital of Oyo State in the southwest of Nigeria. The findings reveal that the majority of these widows are middle aged, between 35 and 55 years of age, with little or no prospect of remarriage. Almost half of them had only primary education and are of low professional status; 48% of them earn very low salary, and had a high parity, having 5 or more children. The problems identified by these widows in order of priority include financial/economic hardship (69%), absence of husband's will resulting in the loss of properties to husband's relations (55%), loneliness and depression (41%), poor relationship with in-laws (41%), difficulty in social interaction (21%), and poor housing (17%). Recommendation for alleviating the hardships of widows suggested include encouragement of female education, enhancement of women, economic empowerment, improving availability and effective utilization of family planning services and encouraging men to write their wills early in marriage. Also, through advocacy and public health awareness campaigns, to enlighten the masses about the plight of the widows, in order to eliminate the dehumanizing traditional practices to which Nigerian widows are often subjected. PMID- 12751558 TI - The pattern of malocclusion among orthodontic patients seen in Dental Centre, University College Hospital, Ibadan, Nigeria. AB - The aim of this study was to analyse the malocclusion pattern among patients who presented for treatment in the Orthodontic Unit of the Dental Centre, University College Hospital, Ibadan, as baseline data for proper treatment planning, teaching and further research. A total of 289 subjects aged 5-34 years with mean age of 10.6 +/- 1.5 (S.D.) years were seen. Angle's classification of molar relationships among those seen is as follows: class I - 76.5%, Class II - 15.5% and Class III - 8.0%. There was increased overjet in 16.2% of the patients, reduced overjet in 0.7% while 2.1% had reversed overjet. Other occlusal abnormalities included: increased overbite (3.8%), reduced overbite (1.4%); anterior open bite (5.2%; crossbite (8.4%) and scissorsbite (0.6%). Crowding, spacing and retained primary incisors constituted 29.7%, 1.4% and 40.1%, respectively. Delayed eruption of canine (1.0%), Bimaxillary protrusion (0.7%), incompetent lips (0.7%), supernumerary teeth (0.7%) malformed tooth (0.3%), mandibular deviation on closure (1.0%) and oral habits (4.5%) were other forms of malocclusion diagnosed. Males were found to have significantly more of classes II and III molar relationships than females (P < 0.05). Occurrence of retained primary teeth as well as overjet deviations from normal were significantly higher in females (P < 0.05). No significant sex differences were found in the other occlusal disorders (P > 0.05): The findings were comparable with previous epidemiological surveys in other parts of the country. PMID- 12751560 TI - Oxygenation of blood in varicose veins. AB - The aim of this prospective, controlled study is to compare the oxygen tension of blood from varicose veins with that from both the antecubital vein of the same patients and the normal long saphenous vein of normal controls at high altitude in Abha (8,000 feet above sea level), Saudi Arabia. Forty-two subjects (21 normal controls and 21 with uncomplicated primary varicose veins) had blood samples taken from 41 normal long saphenous veins and 35 varicose veins near the ankle, respectively (a total of 76 lower limbs). Samples were also taken simultaneously from the right antecubital vein in all the subjects. There was no difference in either oxygen tension (pO2) or saturation (sO2) between blood from varicose veins and blood from normal long saphenous veins. However, in patients with varicose veins, both pO2 and sO2 of varicose veins blood were significantly higher than those of arm venous blood (P = 0.009 and P = 0.018, respectively). In normal subjects, blood from normal long saphenous veins had also significantly higher sO2 (P = 0.001) than that from arm veins, but pO2 was not higher. In conclusion, the theory of arteriovenous communication is poorly founded. There must be other more important explanations for the pathogenesis of varicose veins. PMID- 12751559 TI - Assessing cross infection prevention measures at the Dental Clinic, University College Hospital, Ibadan. AB - The dental clinic is an environment where disease transmission occurs easily. Prevention of cross infection in the dental clinic is therefore a crucial aspect of dental practice and dental clinic workers must adopt certain basic routines while practising. This study evaluates basic routines in prevention of cross infection in the dental clinic, University College Hospital Ibadan. The sample comprised 77 dental clinic workers, who completed closed-ended questionnaires. The questionnaires enquired into practices of the workers in the clinic as well as in the laboratories Physical inspection of dental equipment, instruments and materials was also carried out. The results highlight poor compliance of workers, especially the dental surgeons and students, with the hepatitis B vaccination programme of the Hospital. History to identify high risk individuals was often overlooked and was practised by less than 50% of the clinic workers. Barrier technique with the exception of the use of eye shield, was well practised by all the clinic workers. Aseptic technique was well practised in the dental clinic. Inadequate number of dental surgery assistants, faulty sterilizing equipment, poor monitoring of sterilization, coupled with inadequate number of instruments contributed to the poor success of prevention of contamination and instrument processing procedures. Less than 30% of dental surgeons and fewer than 50% of students discarded sharp materials into the yellow/sharp bin. Liquid waste was well disposed off through the drain for onward flow into the sewer, whilst the disposal of solid contaminated waste did not conform to stipulated international standard. The study found that successful infection control in the dental clinic was highly dependent on the dental surgery assistants, because highly technological equipment were lacking. The management/administration also plays an important role in the number of physical and human resources available and in the overall surveillance of nosocomial infections. PMID- 12751561 TI - Knowledge, perception and practice with regards to occupational risks of HIV/AIDS among nursing and midwifery students in Ibadan, Nigeria. AB - Nursing and midwifery students constitute a subgroup of health care providers exposed to occupational risk of HIV infection became of direct contact with blood and body fluids during clinical practice. The occupational risk faced by these students is of serious concern, given their limited experience and proficiency in nursing care skills. This study was carried out to provide important baseline data about knowledge, perception and practice with regards to occupational risks of HIV/AIDS among 359 nursing and 120 midwifery students (N = 479) in Ibadan, Nigeria. The study utilized both qualitative and quantitative methods to gather information from nursing and midwifery students. The result showed a poor knowledge of WHO recommended universal precaution for preventing HIV transmission among students in all the schools. Students of the schools of midwifery performed more highrisk occupational practises compared to those in the nursing schools, but received less supervision. PMID- 12751562 TI - CD4+ and CD8+ lymphocytes and clinical features of HIV seropositive Nigerians on presentation. AB - Eighty of 200 HIV seropositive patients admitted in the medical wards of Ahmadu Bello University Teaching Hospital, Zaria from year 1995 to 1997 were studied on presentation and compared to 40 age and sex matched controls. The main clinical features observed included weight loss, pyrexia, diarrhoea, lymphadenopathy, anaemia and pruritic dermatosis. Sixty-two of the 80 patients (73.2%) presented at stages 3 and 4 of WHO Clinical and Laboratory staging. Thirty (30) percent of these patients died between a period of one to four months after presentation. The main diseases complicating HIV infection at presentation of the 80 patients were Mycobacterium tuberculosis infection (30%), acute bacterial infections (with Salmonella typhi, Streptococcus pneumoniae and Staphylococcus aureus) (24%), candidiasis (14%) and Kaposi sarcoma (2%). Seropositivity for HIV types was found to be HIV-I alone in 43.5% of cases; HIV-II alone in 14% and both HIV-I and II in 42% of cases. Risk factors associated with HIV infection were multiple sexual partners (73%), sexually transmitted disease (70%), and unscreened blood transfusion (1%). HIV positive patients had a mean CD4+ T-cells of 0.24 x 10(9) +/- 0.17 which was significantly lower than the mean of 0.6 +/- 0. 17 x 10(9)/L for controls (P < 0.05 students t-test). Thirty (35%) of the patients had CD4+ counts of less than 0.2 x 10(9)/L (200 cells/microl) at presentation. The mean CD3+ lymphocytes count was 0.51 +/- 0.24 x 10(9)/L for patients and 1.04 +/- 0.71 x 10(9)/L for controls. The mean CD8+ lymphocyte count in patients was 0.29 +/- 0.19 x 10(9)/L and 0.44 x 10(9)/L for controls. Both CD3+ and CD8+ lymphocyte populations were statistically lower in patients than controls (P < 0.05). PMID- 12751563 TI - The significance of autonomic symptoms in Nigerian diabetics. AB - Diabetes mellitus is the most common endocrine disorder worldwide and disturbance of autonomic function is a frequent complication. Symptoms of autonomic neuropathy are however non-specific and the aetiology is multifactorial in diabetics. This study was carried out to determine the relationship between symptoms of autonomic neuropathy and pathology of the autonomic nervous system in diabetic patients. A hundred diabetics were studied, 50 with, and 50 without symptoms of autonomic neuropathy. Objective test of autonomic function used were the heart rate responses to deep breathing, to standing and to the Valsava manoeuvre; and the blood pressure responses to standing and to sustained handgrip. There was no significant difference between diabetics with symptoms suggestive of autonomic neuropathy, and those without, with regard to findings on autonomic function testing. While the symptoms were unreliable in determining the presence of autonomic neuropathy, they were significantly related to poor glycaemic control and to peripheral neuropathy (P < 0.01). The symptoms of autonomic neuropathy are non-specific in Nigerian diabetics and may reflect poor glycaemic control rather than autonomic neuropathy. Autonomic function tests should be carried out in diabetics suspected of having autonomic neuropathy. PMID- 12751564 TI - Dentofacial anomalies related to the digit sucking habit. AB - Digit (thumb or finger) sucking is the most common oral habit, frequently seen among young children. With an increase in the prevalence of the oral habit in Nigeria in recent years there should be an expected increase in the dentofacial effects of the habit. In this study 81 children, 29 males and 52 females aged 3 16 years were examined. Each child was still actively engaged in the digit sucking habit. Subjects were divided into 3 age groups--3-6 years, 7-10 years and 11 years and above. The dentofacial effects of the habit on each subject were assessed. Increased overjet was observed in 63-705 of the children in the different age groups while the occurrence of anterior open bite ranged from 33.3% to 80% declining in frequency with increase in age. Unilateral posterior crossbite was observed in 8.65 of children while no case of bilateral crossbite was observed. Lip incompetence occurred in 51.8% of children examined, occurring most frequently in the oldest age group. Class 2 skeletal pattern was observed in 22.2% of the sample population. Results show that malocclusion is a frequent result of digit sucking especially when prolonged. There is a need to increase social awareness of the detrimental effects of this habit and if necessary offer alternative non-nutritive sucking methods. PMID- 12751565 TI - Pattern of sexually transmitted diseases among commercial sex workers (CSWs) in Ibadan, Nigeria. AB - The purpose of this study was to determine the pattern of STDs among commercial sex workers (CSWs) in Ibadan, Nigeria. The subjects were 169 CSWs randomly selected from 18 brothels, majority of who were examined and investigated in their rooms. Another 136 women without symptoms who visited the special treatment clinic, University College Hospital, Ibadan were selected as a normal control group. Vaginal candidiasis was the most common STD diagnosed in both CSWs and the control group. The other STDs in their order of frequency were HIV infection 34.3%, non-specific vaginosis 24.9%, trichomoniasis 21.9% and gonorrhoea and "genital ulcers" had an incidence of 16.6% each. Other important conditions were tinea cruris 18.9%, scabies 7.7% genital warts 6.5% and 4.1% of them had syphilis sero-positivity. All the 13 CSWs that had scabies, the 4 (36.4%) with genital warts and the 19 (67.9%) with "genital ulcers" had HIV infection. While there was no significant difference between the CSWs with vaginal candidiasis, gonorrhoea, trichomoniasis and the control group, the HIV positivity was significantly higher (P < 0.001) in CSWs than in the control subjects. These findings suggest that women who exchange sexual services for money can no longer be ignored, and should therefore be identified and made to participate in STD prevention and control programmes. PMID- 12751566 TI - The relationship between juvenile and non-juvenile periodontitis, ABO blood groups and haemoglobin types. AB - This study was carried out to investigate the relationship between juvenile and non-juvenile peridontitis (JP, non-JP), ABO blood groups and haemoglobin type. The heamoglobin electrophoresis was determined by routine technique using cellulose acetate paper and tris buffer at pH 8.5. Tile blood grouping was carried out on all specimens. Forty Nigerian adolescent individuals were investigated, twenty of which were diagnosed as having JP while the remaining 20 were diagnosed a having plaque-induced chronic periodontitis (non JP). This latter group was used as the control group. All the JP patients were either of blood group B/AB, rhesus positive while the non-JP subjects had B rhesus positive/negative, O rhesus positive/negative or AB rhesus positive. The differences between the results of the test and the control groups were statistically significant P < 0.05. All the forty subjects (JP and non-JP) had the haemoglobin type A and none of them exhibited the S and C haemoglobin types. There is a need to further investigate the relationship between juvenile periodontitis, ABO blood group and the common haemoglobin types (A, AS, S, C, and SS) at molecular level. PMID- 12751567 TI - Glucose and insulin responses in offspring of Nigerian Type 2 diabetics. AB - Type 2 diabetes mellitus has a strong genetic basis by a concordance rate ranging between 60 and 90% in monozygotic twins. Glucose and insulin responses to an oral glucose load were measured in 52 offspring of Nigerian Type 2 diabetics and 50 control subjects selected to achieve a similar distribution of age and gender. All subject studied were glucose tolerant. In comparison with control subjects, offspring of Type 2 diabetics had a significantly higher mean (SD) (i) fasting plasma glucose level [69.2 (13.0) mg/dl vs. 62.2 (7.6) mg/dl; P = 0.0012] (ii) fasting plasma insulin level [26.6 (15.4) microIU/ml vs 14.8 (6.8) microIU/ml; P < 0.0001] (iii) 2 hours post glucose load plasma insulin level [59.8 (33.9) microIU/ml vs. 40.9 (24.2) microIU/ml; P = 0.0028]. The mean (SD) 2-hour post glucose load plasma glucose level did not differ significantly between both groups of subjects [92.8 (23.8) mg/dl vs. 85.5 (21.3) mg/dl; P = 0.11]. Further multiple regressional analysis showed that the differences in fasting plasma insulin and 2-hour post glucose load insulin observed were only accounted for by the presence of a parental history of diabetes and were not influenced significantly by BMI, waist and hip circumferences. This study shows that offspring of Nigerian Type 2 diabetics have hyperinsulinaemia, despite being glucose tolerant and this supports the insulin-resistance hypothesis for Type 2 DM. This implies that they are at a greater risk for developing diabetes mellitus and are therefore an important group for the primary prevention of Type 2 DM. PMID- 12751568 TI - Post-measles pneumomediastinum and subcutaneous emphysema in malnourished children. AB - One hundred and seventy-two children with complicated measles were studied clinically andeadiologically for the presence of post-measles pnemomediastinum and subcutaneous emphysema. Eleven cases (6.4%) were found to have this complication. Protein energy malnutrition was an association in 81.8% of these patients. The condition was severe and fatal in one patient, while emergency tracheostomy was needed in another patient who had upper air way obstruction due to gross subcutaneous emphysema. We suggest close observation and early intervention in patients with severe and rapidly progressive subcutaneous emphysema, pneumomediastinum, air block and whenever there is a jeopardy to the cardiovascular system. PMID- 12751569 TI - Infertility: a sociological analysis of problems of infertility among women in a rural community in Nigeria. AB - This study highlighted the problems of infertility among-women in Ilora rural community. The result revealed that in spite of high fertility rate and high rate of pregnancy wastage, 8.7% out of the 400 women studied were found to be infertile. It is interesting to note that in a strictly polygamous environment, almost half of the women suffering from infertility were found to be the only wife of their respective husbands. The African concept as to the solution to the problems was also discussed, and highlighted in relation to awareness of possible hospital treatment of their conditions. PMID- 12751570 TI - The deficient perineum: oblique presentation of a clinically obvious anomaly. AB - The deficient perineum is a recognized anatomical defect attributable essentially to the trauma of vaginal delivery. We studied, retrospectively, 64 women with the deficient perineum who presented in our unit for other reasons. All were parous and had previous vaginal delivery. Eighty-four percent delivered recently outside a hospital. Presumed marital disharmony occurred in 45% and may have been profound. Perineorrhaphy in the women seen, had salutary effects. PMID- 12751571 TI - Short-term haemodialysis in pregnant patients with acute renal failure: a report of two cases. AB - Haemodialysis in pregnancy is not common although, successful dialysis in pregnancy have been reported. It has also been found to improve survival of both mother and child especially, in patients with chronic renal failure, with pre term labor being a common occurrence. Out of the 2,995 patients that were dialyzed at the University College Hospital, Ibadan in the last 10 years, only 2 of the patients were pregnant and both of them had acute renal failure. We present here the two cases, which represents our experience at the University College Hospital, Ibadan, Nigeria. PMID- 12751572 TI - Sixty-one day twin to twin birth interval in a low technology setting. AB - Multiple pregnancy, whether spontaneous or from artificial reproductive techniques, is regarded as high risk. Limiting the twin-twin birth interval to within 30 minutes has been widely practised. However, conservative management of the retained live, immature second twin may be worthwhile. We report a case lasting sixty-one days in a low technology setting. PMID- 12751573 TI - Rigid retrograde endoscopy under regional aneasthesia: a novel technique for the early realignment of traumatic posterior urethral disruption. AB - Traumatic disruption of the posterior urethra usually occurs in association with pelvic fractures and may result in significant morbidity. The management of this injury remains difficult and controversial. Recently, early restoration of urethral continuity in these patients using either both antegrade and retrograde cystoscopy (with or without fluoroscopy), or flexible retrograde urethroscopy alone under general aneasthesia, has been reported with good results. These procedures have been proposed as an improvement over the traditional teaching of placement of a suprapubic catheter followed by delayed open repair (urethroplasty). We now describe a novel method of restoration of urethral continuity by rigid retrograde endoscopy alone under caudal aneasthesia in the early post-trauma phase. This procedure can be carried out in an outpatient endoscopic suite with standard (endoscopic)optical urethrotomy equipment and is suitable for most patients with prostatomembranous urethral disruptions. A 'high riding' prostate is however a relative contraindication for this procedure. To date, we have carried out this procedure successfully in 4 of 5 patients with traumatic prostato-membranous disruption (a success rate of 80%). We recommend that early retrograde rigid endoscopic realignment under regional analgesia should be considered as a management option in patients with traumatic disruption of the posterior urethra. PMID- 12751574 TI - Isolated umbilical endometriosis--a rare finding. AB - A 32 yr old multiparous Nigerian woman presented with histological confirmed umbilical endometriosis. There was no evidence of endometrotic deposits elsewhere. Apart from cyclical bleeding from the umbilicus, she was relatively free of any of the other signs and symptoms of Endometriosis. She had excision of the endometeriotic lesion and responded favourably to treatment. A high index of suspicion is required in making a diagnosis of endometriosis in remote extra pelvic sites especially with little or no characterisitic pointers to the disease. PMID- 12751575 TI - HIV-assocated lymphoma: a case report. AB - We report the case of a 3(1/2) year old boy who presented with sudden onset of headache. Fever and swelling of the left eye. He had complete opthalmoplegia of the left eye and 6th cranial nerve paralysis in the right eye. He was thought to have cavenous sinus thrombosis but CT findings suggestive of lymphoma led to the correct diagnosis of HIV associated Lymphoma It view of the rising incidence of HIV infection and the protein clinical manifestations, it is advised that all patients with disseminated tumour masses should be screened for the HIV virus, and CT examination should be made available to patients. PMID- 12751576 TI - Using flight simulators aboard ships: human side effects of an optimal scenario with smooth seas. AB - INTRODUCTION: The U.S. Navy is considering placing flight simulators aboard ships. It is known that certain types of flight simulators can elicit motion adaptation syndrome (MAS), and also that certain types of ship motion can cause MAS. The goal of this study was to determine if using a flight simulator during ship motion would cause MAS, even when the simulator stimulus and the ship motion were both very mild. METHODS: All participants in this study completed three conditions. Condition 1 (Sim) entailed "flying" a personal computer-based flight simulator situated on land. Condition 2 (Ship) involved riding aboard a U.S. Navy Yard Patrol boat. Condition 3 (ShipSim) entailed "flying" a personal computer based flight simulator while riding aboard a Yard Patrol boat. Before and after each condition, participants' balance and dynamic visual acuity were assessed. After each condition, participants filled out the Nausea Profile and the Simulator Sickness Questionnaire. RESULTS: Following exposure to a flight simulator aboard a ship, participants reported negligible symptoms of nausea and simulator sickness. However, participants exhibited a decrease in dynamic visual acuity after exposure to the flight simulator aboard ship (T[25] = 3.61, p < 0.05). Balance results were confounded by significant learning and, therefore, not interpretable. DISCUSSION: This study suggests that flight simulators can be used aboard ship. As a minimal safety precaution, these simulators should be used according to current safety practices for land-based simulators. Optimally, these simulators should be designed to minimize MAS, located near the ship's center of rotation and used when ship motion is not provocative. PMID- 12751577 TI - Measurement of brain electrical impedance: animal studies in rheoencephalography. AB - INTRODUCTION: In spite of the known importance of cerebral blood flow (CBF) monitoring for aviation, spaceflight, military and emergency medicine, and neurosurgical intra- and postoperative monitoring, there is no standard noninvasive technique for continuous CBF monitoring. One potential method for this purpose is the electrical impedance technique, called rheoencephalography (REG). The development of improved electronics and computation tools has done much to overcome the difficulties of REG measurement. REG technology now has possibilities for application to the fields mentioned above. HYPOTHESIS: Our hypothesis was that REG would reflect CBF changes. METHODS: Three experimental studies were undertaken to further define in vivo (rat, pig) CBF measurements by analysis of REG pulse waves. CO2 inhalation (4-20%), brain electrical stimulation, and aorta compression (5 min) were the applied CBF manipulations. In the case of aorta compression, global CBF was measured by REG, and local CBF by the laser Doppler method. Data were digitized and processed off-line. RESULTS: During CO2 inhalation and electrical stimulation of the brain, REG amplitude increased, indicating increased cerebral fluid volume. A linear relationship was established between CO2 concentration and REG peak amplitude (correlation coefficient: 0.88, p = 0.05), and the ascending portion of the curve (0.88, p = 0.05). During aorta compression, systemic arterial pressure increased (p = 0.008), and REG amplitude decreased (-23.75%, p = 0.01). CONCLUSION: These studies have confirmed the REG amplitude changes during known CBF manipulations. The difference between local and global CBF response demonstrated CBF autoregulation and heterogeneity. Together, these studies indicate the usefulness and potential benefit of computerized REG monitoring for the above-mentioned fields. PMID- 12751578 TI - Lumbar spine disc heights and curvature: upright posture vs. supine compression harness. AB - INTRODUCTION: Spinal lengthening in microgravity is thought to cause back pain in astronauts. A spinal compression harness can compress the spine to eliminate lengthening but the loading condition with harness is different than physiologic conditions. Our purpose was to compare the effect of spine compression with a harness in supine position on disk height and spinal curvature in the lumbar spine to that of upright position as measured using a vertically open magnetic resonance imaging system. METHODS: Fifteen healthy subjects volunteered. On day 1, each subject lay supine for an hour and a baseline scan of the lumbar spine was performed. After applying a load of fifty percent of body weight with the harness for thirty minutes, the lumbar spine was scanned again. On day 2, after a baseline scan, a follow up scan was performed after kneeling for thirty minutes within the gap between two vertically oriented magnetic coils. Anterior and posterior disk heights, posterior disk bulging, and spinal curvature were measured from the baseline and follow up scans. RESULTS: Anterior disk heights increased and posterior disk heights decreased compared with baseline scans both after spinal compression with harness and upright posture. The spinal curvature increased by both loading conditions of the spine. DISCUSSION: The spinal compression with specially designed harness has the same effect as the physiologic loading of the spine in the kneeling upright position. The harness shows some promise as a tool to increase the diagnostic capabilities of a conventional MR system. PMID- 12751579 TI - Optokinetic stimulation in microgravity: torsional eye movements and subjective visual vertical. AB - INTRODUCTION: Microgravity provides unique sensory inputs to the vestibular and oculomotor systems. We sought to determine the effects of long-term spaceflight on sensing of spatial orientation. METHODS: Two cosmonauts participated in experiments on human vestibulo-visual interactions during a long-term mission (178 d) in the MIR station in 1995. During circular optokinetic stimulation (OKS) the tonic torsional eye position (torsional beating field, TBF) and the subjective visual vertical (SVV) were recorded on several days of the space mission as well as pre- and post-flight. A reference data set was obtained from healthy subjects on Earth, in whom the TBF was measured in upright and in prone positions. RESULTS: Neither cosmonaut showed changes in the SVV or the TBF values during the first days in microgravity. On flight day 149, cosmonaut A showed an increase of both values, which continued to rise by 4- and 10-fold until the end of the flight (TBF: 8.1 degrees; SVV: 216.8 degrees). This cosmonaut reported that the increase was accompanied by a loss of spatial orientation. In contrast, cosmonaut B's values remained at pre-flight levels (TBF: 1.6 degrees; SVV: 4.4 degrees). Post-flight values of the TBF did not significantly differ from pre flight values for either cosmonaut. Subjects showed an increase of the TBF by more than a factor of 2 in prone position (range -7.7 degrees to +10.2 degrees) compared with upright position (range -3.7 degrees to +3.4 degrees). CONCLUSIONS: Pre-flight, post-flight and during the first part of the flight, both cosmonauts exhibited values similar to those of normal subjects in an upright position. The increased TBF values of cosmonaut A from flight day 110 on were within the range of the normal subjects in prone (face-down) position, when the gravity vector cannot be used to stabilize the TBF against the rotating stimulus (the axis of rotation is parallel to the gravity vector). The increasing deviations of cosmonaut A's SVV values in-flight suggest the presence of an internal body reference system, which weakened throughout the flight and thus lost its stabilizing effect. PMID- 12751580 TI - Effect of hypoxia on preferred hand temperature. AB - INTRODUCTION: Selection of preferred skin temperature is a consequence of complex central neural integration of thermoafferent information in the polysynaptic pathway for behavioral thermoregulation. The present study tested the hypothesis that hypoxia alters behavioral temperature regulation. METHODS: Eight male volunteers participated in two separate trials. In each trial they were immersed in 28 degrees C water for 90 min, and inspired either room air (air trial) or a hypoxic gas mixture (hypoxia trial; 11.5% O2/88.5% N2). Their left hand was placed in a separate bath perfused with water. At regular intervals they were instructed to adjust the temperature of the water perfusing the hand bath to a comfortable temperature, defined as the 'preferred hand temperature' (Tpref). At minute intervals, esophageal (Tes), rectal (Tre), and skin (Tsk) temperatures, along with heat flux from the skin (Q) and ventilation (VI) were recorded. The observed responses were compared with a one-way ANOVA. RESULTS: With the exception of VI, there were no significant differences in any of the recorded variables between the two conditions. During the immersion, Tes and Tre decreased by 1.0 and 0.7 degrees C, respectively, in the air trial, and by 0.8 and 0.7 degrees C, respectively, in the hypoxia trial. Concomitant with decreases in core temperature, there was a significant increase (p < 0.05) in Tpref of 2.1 degrees C in the air trial and 2.4 degrees C in the hypoxia trial. The difference in Tpref between the air and hypoxia trials was not significant. DISCUSSION: Assuming that the selection of preferred hand temperature represents a behavioral thermoregulatory response, the present results suggest that moderate normobaric hypoxia does not affect human behavioral thermoregulation. PMID- 12751581 TI - Neck muscle activity in helicopter pilots: effect of position and helmet-mounted equipment. AB - BACKGROUND: Helicopter pilots usually work in unfavorable ergonomic positions, often with bulky head-worn equipment during flying missions. The purpose of this study was to evaluate and compare immediate muscle response in the dorsal neck muscles to different positions with a variety of head-worn equipment. METHODS: Fourteen healthy male helicopter pilots volunteered for this study. EMG activity in the upper and lower dorsal neck muscles and the trapezius muscle was measured in a laboratory situation for 5 s in different sitting positions (neutral, trunk inclined 20 degrees, neck flexed 20 degrees), including registration of a 30 degrees left and right rotation in every position; all measurements were performed while wearing a helmet, a helmet and night vision goggles (hNVG), and a helmet, night vision goggles, and counterweight (hCW), in random order. RESULTS: There was significant higher EMG activity in the upper neck with hNVG and hCW than with the helmet only when comparing the mean activity level of all positions. However, there was no significant difference in EMG activity between any variations of head-worn equipment when comparing activity levels during each position separately. In the upper and lower neck, respectively, there was significantly higher muscle activity during the ipsilateral rotated positions plus neck flexion and trunk inclination than in most other positions. CONCLUSION: The increased load caused by different positions seems to have a greater influence on muscle activity than the increased load of the head-worn equipment, which must be considered when designing helicopter work-places. PMID- 12751582 TI - Blood gases, hematology, and renal blood flow during prolonged mountain sojourns at 3500 and 5800 m. AB - BACKGROUND: This study mainly focused on renal blood flow, hematological parameters, blood gases, and blood pH, which are affected on exposure to moderate (3500 m) and extreme altitudes (5800 m) in sea level residents. HYPOTHESIS: Acute and prolonged exposure to high or extreme altitude may cause pathophysiological changes in kidney and renal plasma/blood flow, leading to retention of fluids in the tissues. Combined with the decreased availability of oxygen to the tissues, these may be responsible for high altitude maladies. METHOD: Fifteen male sea level (SL) volunteers, 22-25 yr old, were studied for blood gases, blood viscosity, hematocrit, hemoglobin concentration, and effective renal blood/plasma flow at Delhi (260 m), 3500 m (60 d stay), 5800 m (70 d stay), and 7 d after return to SL. RESULTS: Compared with SL, a significant increase from 7.34 to 7.43 (p < 0.01) in blood pH was observed at 3500 m that remained significantly increased at 5800 m. PO2 was about 39% less at 5800 m than at SL. PCO2 reduced significantly from 42.07 to 28.05 mm Hg on return from 5800 m to SL. The blood viscosity increased significantly (38%) at 5800 m and decreased significantly by 38% (p < 0.01) after return to SL. The effective renal plasma flow reduced significantly (p < 0.01) from 615.6 at SL to 381.5 ml x min(-1) x 1.73 m(-2) at 5800 m. CONCLUSION: The study suggests a crucial role of renal function in the acclimatization process; renal function also appears to be one factor by which the body protects itself against severe hypoxia. PMID- 12751583 TI - Training improves divers' ability to detect increased CO2. AB - BACKGROUND: Elevated arterial PCO2 (hypercapnia) is a known risk in diving with closed circuit breathing apparatus. In a retrospective study, we determined CO2 retention and the ability to detect CO2 in novice divers who were either CO2 recognition-trained subjects (TS) or untrained subjects (UTS). METHODS: Ventilatory and perceptual responses to variations in inspired CO2 (range 0-5.6 kPa, 0-42 mm Hg) during moderate exercise were assessed in novice Israeli Navy divers on active duty. Tests were carried out on 231 TS and 213 UTS. RESULTS: The minimal mean inspired PCO2 that could be detected was 4.8 +/- 1.6 kPa (36 +/- 12 mm Hg) in UTS and 2.9 +/- 0.7 kPa (22 +/- 5 mm Hg) in TS (p < 0.0001). No significant changes were found in PETCO2 between the two groups during exposure to a PICO2 of 5.6 kPa (42 mm Hg). There were 46 TS who were found to be CO2 retainers (more than +1 SD above the mean) and 19 were classified as poor detectors (more than +1 SD above the mean). Seven subjects exhibited both traits. During actual oxygen diving performed later by this group, the only four cases of CNS-oxygen toxicity were among those seven subjects (p < 0.01). CONCLUSIONS: We conclude that CO2 recognition training improves the diver's capability to detect CO2. We suggest that a diver who is both a poor CO2 detector and a CO2 retainer will be prone to CNS-oxygen toxicity. PMID- 12751584 TI - The performance of color deficient individuals on airfield color tasks. AB - BACKGROUND: The pseudo-isochromatic plate (PIP) test (e.g., Ishihara test) is the clinical test commonly used to assess color vision. Upon failure of this test, candidates are typically reassessed using the Farnsworth Lantern (FALANT) test to determine their fitness for occupations which require normal color vision. We were interested in determining to what extent clinical tests can predict real life color naming performance, particularly in the context of "airside drivers" (any airport vehicle operators who drive on the airfield). METHODS: There were 24 male subjects with a color vision deficiency, as defined by the Ishihara test, who participated in this study. They were further assessed using the D-15 and lantern color vision tests. All subjects then participated in two separate color naming tasks. These tasks consisted of naming surface colors and colored-lights of the type used on the airfield of the Hong Kong International Airport (HKIA). RESULTS: Of the 24 subjects, 15 failed both D-15 and the FALANT tests. Out of these 15 subjects, 8 also failed the naming tasks. The FALANT test showed very good agreement (87.5%) with the Ishihara test. Similar to the Ishihara, FALANT tests had 100% sensitivity in identifying the subjects who failed the naming tasks. The agreement between the Ishihara and D-15 tests was 62.5%. DISCUSSION: In common with previous studies, our results show that clinical tests cannot predict accurately who will fail color naming tasks of the type normally encountered in the real-life work environment. The high false positive values of the clinical tests in relation to color naming tasks suggest that people with color deficiency may not be given a fair opportunity to demonstrate their true ability in performing the task. PMID- 12751585 TI - New test to assess pilot's vision following refractive surgery. AB - BACKGROUND: All forms of corneal refractive surgery can sometimes cause an increase in optical aberrations and scattered light, which can affect visual performance. The purpose of this study was to develop a suitable test that was sensitive to retinal image degradation in subjects who have undergone excimer laser refractive surgery and that was also relevant to visual demands in commercial aviation. METHODS: Assessment of the visual environment and the tasks involved in piloting a commercial aircraft formed the basis for the selection of the test parameters. The new contrast acuity assessment (CAA) test covers a functional visual field of +/-5 degrees and is based on minimum spatial vision requirements for commercial pilots. RESULTS: Data measured in 100 normal subjects were used to define the 'standard normal observer' and the range of variation for the parameters of the test. This approach makes it possible to quickly establish whether a given subject's performance falls within the range of the standard normal observer. The test is also administered under low ambient illumination since flying at night involves mesopic levels of light adaptation when the pupil size is large and the effects of aberrations and scattered light are therefore more pronounced. CONCLUSION: The results of the test are simple to interpret and reveal visual performance that falls outside the normal range as a result of either significant degradation of retinal image quality (caused by increased aberrations and scattered light) or abnormal processing of visual information in the retina and/or the visual pathway. PMID- 12751586 TI - Medical risk factors in fatal military aviation crashes: a case-control study. AB - BACKGROUND: Periodic medical examinations are the daily work of the flight surgeon. Their immediate impact on flight safety, however, has not been evaluated. This case-control study was done to ascertain whether, among German military pilots, differences exist in the results of periodic medical examinations that were associated with a higher odds of being involved in a fatal aircraft mishap. METHODS: Participants were 146 German military pilots who died in air crashes and 292 controls. Cases and controls were matched 1:2 by aeronautical confounders (age, type of aircraft aeronautical experience, and membership in the air force, army, or navy). Data source was the central register of the German Air Force Institute of Aviation Medicine; data were obtained from periodic medical examinations and included physical examination and laboratory data. Odds ratios were calculated by conditional logistic regression analysis. RESULTS: Descriptive statistics showed no distinct difference between cases and controls in most of the parameters considered except for total serum bilirubin, but multiple conditional logistic regression showed no remarkably different odds for any of the parameters tested. CONCLUSION: In aircrew who pass all the criteria for fitness to fly there are no important medical risk factors for fatal air crashes that can be detected by periodic medical examinations. PMID- 12751587 TI - Epidemiology of U.S. Army cold weather injuries, 1980-1999. AB - BACKGROUND: Cold weather injuries (CWI) are of great military concern due to their wide-ranging impact on military readiness. Previous short-term studies have identified CWI to be more prevalent in African-Americans, infantrymen, and lower ranking soldiers. The purpose of this cross-sectional study was to determine the occurrence of CWI hospitalizations in the U.S. Army from 1980 to 1999, and to identify possible trends, high-risk groups and/or activities. METHODS: The U.S. Army Research Institute of Environmental Medicine Total Army Injury and Health Outcomes Database was searched for hospitalizations with ICD-9-CM codes for frostbite, hypothermia, immersion foot, chilblains, and other. Information concerning each soldier included: gender, age, ethnicity, rank, occupation, type of injury, home of record, duty station, principle diagnosis, trauma code, and cause of injury. Data was available on the demographic composition of the Army, by year, and was used as the denominator when calculating the frequency of occurrence. RESULTS: During the study period there were 2143 hospitalizations due to CWI. African-American men and women were injured approximately 4 times and 2.2 times as often as their Caucasian counterparts, respectively. Trauma and cause of injury codes indicate that about 80% of all CWI hospitalizations result on-duty and during organized training. The yearly rate of CWI hospitalization has declined from 38.2/100,000 in 1985 to 0.2/100,000 in 1999. CONCLUSIONS: Our data are consistent with previous research concerning the increased rate of CWI among African-Americans though further investigation appears warranted. The occurrence of most CWI during on-duty training suggests preventability. The decline in the overall rate of CWI hospitalizations is multifactorial. PMID- 12751588 TI - Pilots with vasovagal syncope: fit to fly? AB - Two pilots who had experienced vasovagal syncope were grounded by the aeromedical service. Pilot A had experienced three episodes of syncope in medical settings, none during flight. Pilot B had experienced four episodes of syncope in emotional/medical settings, one during flight. Whether a pilot who experienced one or more episodes of vasovagal syncope is declared fit to fly now depends on the number of episodes experienced. We propose that pilots should be assessed individually. Certainty of the diagnosis of vasovagal syncope, the chance and predictability of recurrences during flight, and the possibility of effective therapy should be assessed. Chance of recurrence during flight is low when the triggering factor is known and avoidable. Pilots with syncopal episodes in predictable (e.g., medical) situations, with clear prodromal symptoms and/or effective therapy, should be declared fit to fly. A symptom-free period and/or restriction to fly 'as or with a co-pilot' can be considered. PMID- 12751589 TI - Intercultural crew issues in long-duration spaceflight. AB - Before long-duration flights with international crews can be safely undertaken, potential interpersonal difficulties will need to be addressed. Crew performance breakdown has been recognized by the American Institute of Medicine, in scientific literature, and in popular culture. However, few studies of human interaction and performance in confined, isolated environments exist, and the data pertaining to those studies are mostly anecdotal. Many incidents involving crew interpersonal dynamics, those among flight crews, as well as between flight crews and ground controllers, are reported only in non-peer reviewed books and newspapers. Consequently, due to this lack of concrete knowledge, the selection of astronauts and cosmonauts has focused on individual rather than group selection. Additional selection criteria such as interpersonal and communication competence, along with intercultural training, will have a decisive impact on future mission success. Furthermore, industrial psychological research has demonstrated the ability to select a group based on compatibility. With all this in mind, it is essential to conduct further research on heterogeneous, multi national crews including selection and training for long-duration space missions. PMID- 12751590 TI - Cases from the aerospace medicine residents' teaching file. Penetrating chest trauma from a home accident: rapid response saves a pilot. AB - Penetrating cardiac chest trauma can be associated with a high degree of mortality. Prognostic factors that favor a high survival rate include OR thoracotomy vs. ER thoracotomy, prompt resuscitation, fast transport, urgent diagnostic study, and immediate surgery. Cardiac tamponade, in some cases, may lessen the amount of hemorrhage. Reported is a case of penetrating cardiac trauma in an aviator. With a high degree of suspicion and rapid transport, early intervention was possible resulting in a favorable outcome. The patient, a United State Marine Corps Harrier pilot, returned to flight status with a waiver. PMID- 12751591 TI - Central serous retinopathy. PMID- 12751592 TI - Antibiotics for URIs. PMID- 12751593 TI - Head pediculosis. PMID- 12751594 TI - This month in aerospace medicine history--May 2003. PMID- 12751595 TI - Facial recognition technology (FERET). PMID- 12751596 TI - U. S. Naval Aerospace Physiology Program. PMID- 12751597 TI - Current indications for transurethral resection of the prostate and associated complications. AB - Transurethral resection of the prostate (TURP) is the most common surgical procedure for relieving symptoms of benign prostatic hyperplasia. Here, we report our experience of current indications for TURP and their associated outcomes at Kaohsiung Medical University Hospital (KMUH). A total of 111 patients who underwent TURP at KMUH between May 2000 and December 2001 were included in this retrospective review. For each patient, the surgical indication was categorized into acute urinary retention, chronic complications (including renal impairment, recurrent urinary infection, bladder stone/diverticulum, post-void residue, and recurrent hematuria), and symptomatic prostatism. Thirty-five patients (31%) had acute urinary retention, 28 (27%) had chronic complications, and 48 (42%) had symptomatic prostatism. Most patients chose TURP only when medical treatment had failed to relieve symptoms, no matter what category they belonged to. Patients with acute urinary retention and chronic complications had larger prostates (p = 0.002) and more tissue resected (p = 0.05) than those with symptomatic prostatism. Patients with acute urinary retention seemed to be at greater risk of postoperative complications such as recurrent urinary retention and urinary tract infection. We suggest that urodynamic study may be necessary to rule out concomitant bladder dysfunction before surgery and that adequate prophylactic antibiotic treatment be used to decrease the risk of urinary tract infection during or after TURP, especially when pyuria is noted preoperatively in patients with acute urinary retention. PMID- 12751598 TI - Intraductal papillary mucinous tumor of the pancreas: computerized tomography and magnetic resonance imaging features. AB - The aim of this study was to analyze the computerized tomography (CT) and magnetic resonance imaging (MRI) features of intraductal papillary mucinous tumor (IPMT) of the pancreas. The cases of eight patients with pathologically proven IPMT (1 papillary hyperplasia, 7 adenocarcinoma) of the pancreas were retrospectively reviewed. There were five men and three women with ages ranging from 42 to 82 years. Imaging studies included six thin-section dynamic CT scans, seven MRI scans, one MR cholangiopancreatography scan, and two endoscopic retrograde cholangiopancreatography scans. There was only one benign IPMT, which presented as a unilocular cyst in the pancreatic body with no mural nodules and no dilatation of the main pancreatic duct (MPD). All seven patients with malignant IPMT had multilocular cysts with papillary projections in the pancreatic head and/or uncinate process accompanied by dilated MPD (5 diffuse, 2 segmental). Communication between the cystic lesions and the MPD were evident in all seven patients. One patient had small mural nodules in the branch ducts of the pancreatic body and five had a bulging papilla with a patulous orifice. A mass effect resulting in biliary obstruction was shown in one patient. One patient had a ruptured cyst with mucin leakage into the right anterior pararenal space following sono-guided aspiration. In conclusion, the main imaging feature of IPMT in our patients was a multilocular cyst with papillary projections located in the pancreatic head and uncinate process. Although CT and MRI cannot differentiate mucin content from pancreatic juice, communication between the cystic lesion and the dilated MPD and a bulging papilla with a patulous orifice are characteristics of IPMT. PMID- 12751599 TI - Effect of filmless imaging on utilization of radiologic services with a two stage, hospital-wide implementation of a picture archiving and communication system: initial experience of a fee-for-service model. AB - A medium-sized general hospital using a fee-for-service model implemented a hospital-wide picture archiving and communication system (PACS) in two stages. This study evaluated the reporting time with filmless operation and the effect of filmless imaging on referring physicians' use of the radiologic service before and after completion of the second stage of PACS implementation. The relationship between the total number of hospital patients and the number of radiologic department patients was also evaluated. All sample images were retrieved from the PACS. All corresponding reports except for one for a computerized tomography study were available. The median reporting time for different studies performed during working hours was less than 2 hours. There was a significantly positive and linear relationship (p < 0.01) between the total number of hospital patients and the number of radiologic department patients after hospital-wide implementation of PACS. We conclude that the fee-for-service model had no negative impact on referring physicians' use of radiologic services in a filmless hospital. PMID- 12751600 TI - Topographic study of extracted molars with advanced furcation involvement: furcation entrance dimension and molar type. AB - This study investigated the topography of the furcation entrance dimension (FED) on molars with advanced furcation involvement (FI). The sample pool consisted of 169 maxillary and mandibular molars from a group of 165 individuals with severely advanced periodontal destruction. The subjects included men and women aged 24 to 84 years (mean, 47.8 +/- 7.2 years). The FEDs of the maxillary buccal, mesial, and distal surfaces as well as the mandibular buccal and lingual surfaces were measured under a stereomicroscope and clarified into grades I (FED < 0.55 mm), II (0.55-0.75 mm), and III (> 0.75 mm) using automatic grading system software designed by our research associates. The differences and relationships among molar location, furcation site, and FED grade were analyzed using the chi-square test. There was a significant difference among buccal (BFED), mesial (MFED), and distal (DFED) FEDs in both the maxillary first (chi2(4) = 58.915, p < 0.001) and second (chi2(4) = 66.839, p < 0.001) molars. The relationship between molar type and FED grade was statistically significant for both the BFED (p < 0.001) and the DFED (p < 0.001) of maxillary molars, as well as for both the BFED (p < 0.0001) and LFED (p < 0.0001). The difference in FED grade between the first and second molars was statistically significant in both maxillary (p < 0.001) and mandibular (p < 0.0001) molars. There was a significant relationship between FED grade and tooth location at molars with advanced FI. PMID- 12751601 TI - Hemorrhagic corpus luteum cyst torsion in term pregnancy: a case report. AB - Hemoperitoneum during pregnancy resulting from spontaneous rupture of adnexal torsion is a rare cause of fetal and maternal death. Presenting symptoms include severe abdominal pain, followed rapidly by maternal shock and fetal distress. It is hard to localize the adnexae in advanced pregnancy. Here, we present a case of spontaneous rupture of hemorrhagic corpus luteum cyst torsion that had not been previously diagnosed by ultrasound during term pregnancy. The patient was sent to our emergency room for sudden onset of severe low abdominal pain. Treatment consists of maintenance of adequate circulating intravascular volume and rapid surgical intervention. Preoperative diagnosis of adnexal torsion during term pregnancy is very difficult, although it is always identified during surgery. PMID- 12751602 TI - Bilateral superselective arterial microcoil embolization in post-traumatic high flow priapism: a case report. AB - Priapism is a prolonged penile erection unrelated to sexual stimulation. High flow arteriogenic priapism is uncommon and usually occurs after genitoperineal trauma, which may damage a feeding cavernosal artery, leading to an arteriovenous fistula and, occasionally, to an associated pseudoaneurysm. The defects rarely occur bilaterally. Herein, we report successful treatment of high-flow priapism secondary to a traumatic pseudoaneurysm fed from the bilateral cavernosal artery. Diagnosis was made after cavernosal blood gas analysis, color Doppler ultrasonography, and superselective angiography. Treatment consisted of superselective arterial embolization using metallic microcoils and resulted in simultaneous detumescence of the penis with no complications. The patient regained morning erection on the second postoperative day and erectile function remained normal 8 months after treatment. This case shows that bilateral arteriocavernosal fistulae can be successfully treated with superselective arterial embolization without affecting potency and highlights the importance of warning men about the possibility of developing high-flow priapism following a perineal trauma. PMID- 12751603 TI - Double J stent forgotten for 7 years: a case report. AB - Ureteral stents are an integral part of urologic practices. Nonetheless, stents that migrate, fragment, or are forgotten pose management and legal dilemmas. We report the case of a "forgotten" stent, which migrated upward into the right renal pelvis, concomitant with a ureteral stone. The "forgotten" stent was placed 7 years previously during right ureteral stone treatment. The patient finally underwent nephroscopic stent retrieval and ureteroscopic ureteral stone removal. The final plain X-ray demonstrated no stone fragment or residual double J stent fragment. The patient withstood the procedure well and was free of symptoms thereafter. PMID- 12751604 TI - Management of low-flow priapism using the Winter procedure: a case report. AB - Priapism is a prolonged penile erection that is unrelated to sexual stimulation. Low-flow priapism has been associated with sickle cell disease and other hemoglobinopathies, neoplastic syndrome, anticoagulant therapy, psychotropic medication, and idiopathic causes. We report the successful treatment of idiopathic low-flow priapism using the Winter procedure. Initial treatment consisted of aspiration and intracavernous irrigation with iced saline and a vasoconstrictive agent, but in vain. We then performed the Winter procedure, in which fistulas between the corpora cavernosa and the glans penis were created. This resulted in the simultaneous detumescence of the penis, without complication. The erectile function of the penis was normal 1 year after the procedure. This case shows that idiopathic low-flow priapism can be successfully treated using the Winter procedure when conservative treatment fails. PMID- 12751605 TI - Preliminary study of joint disease in poultry by the analysis of synovial fluid. AB - Samples of synovial fluid and synovial membrane were obtained from the hock joints of several groups of broilers, including lame birds and two strains of broilers raised on different feeding regimens and given different drug treatments (carprofen or placebo). There were more significant differences between the groups on the basis of the analysis of the synovial fluid samples than the synovial membrane samples. Experimental birds fed ad libitum had the highest median red blood cell counts and median ghost cell counts of all of the groups, but there were no differences between the groups in the thickness of the synovial lining cell layer or the degree of cellular infiltrate in the synovial membrane. The synovial fluid from the broilers and lame birds fed ad libitum was more turbid, suggestive of intra-articular pathology, and the large numbers of heterophils in samples from the lame birds indicated an inflammatory arthropathy. The birds fed ad libitum which were treated with carprofen had more cells in the synovial fluid than the birds given the placebo. A large number of the samples of synovial fluid from the ad libitum-fed broilers contained blood. PMID- 12751606 TI - Study of the effect of Bronchipret on the lung function of five Austrian saddle horses suffering recurrent airway obstruction (heaves). AB - The effects of an oral preparation containing an extract of thyme and primula (Bronchipret; Bionorica) on the lung function of five horses suffering heaves were determined in a longitudinal study. The horses accepted the product well. The plasma concentrations of the marker substance, thymol, indicated that at least one of the substances in the extract had been absorbed from the gastrointestinal tract. The compliance, pulmonary pressure and airway resistance of the horses' lungs were all significantly improved after one month of treatment However, the severity of their clinical signs and their arterial oxygen partial pressure had not improved significantly. PMID- 12751607 TI - Serum amyloid A in the serum and milk of ewes with mastitis induced experimentally with Staphylococcus epidermidis. AB - Mastitis was induced experimentally in ewes with Staphylococcus epidermidis, and the concentrations of serum amyloid A (SAA) in milk and serum, and the somatic cell counts and bacteria in the milk were determined for up to 10 weeks in two experiments, each examining five infected and five control ewes. The somatic cell counts peaked eight hours after infection and preceded an increase in SAA in milk. A maximum concentration of 6460 microg/ml SAA was recorded in milk from the infected sheep, compared with a mean concentration of 1.4 microg/ml in the control sheep. The mean peak concentration of SAA in serum (206.8 microg/ml) occurred earlier (one day after infection) than in milk. The serum concentration of SAA in the healthy animals ranged from 0 to 29.4 microg/ml. There was no correlation between the concentrations of SAA in serum and milk. PMID- 12751608 TI - Haptoglobin concentrations in a canine hospital population. PMID- 12751609 TI - Prospective study of the treatment of feline plasmacytic pododermatitis with doxycycline. PMID- 12751610 TI - Intravaginal fibroma in a sheep. PMID- 12751611 TI - Ectoparasite awareness in sheep. PMID- 12751612 TI - Veterinary drugs found in Shropshire. PMID- 12751613 TI - Neuronal degeneration in an Irish moiled bull calf. PMID- 12751614 TI - Over-30-months scheme. PMID- 12751615 TI - Embryo transfer success in the Falkland Islands. PMID- 12751616 TI - Sunburn in Australian men with a history of non-melanoma skin cancer. AB - OBJECTIVE: To identify predictors of recent sunburn in north Australian men with a history of non-melanoma skin cancer (NMSC). METHODS: A survey of men with previous NMSC was conducted (n = 300, response rate 62%). RESULTS: Fifty-four percent of participants reported recent sunburn. Predictors identified included younger age, belief that NMSC is caused by childhood sun exposure, belief that sun protection will not help prevent further NMSC, wearing of casual clothes, and use of shade as the main sun-protection strategy. CONCLUSION: Health promotion messages should emphasize the importance of sun protection throughout life and the use of stringent sun-protection measures. PMID- 12751617 TI - Using Internet-based surveys to reach hidden populations: case of nonabusive illicit drug users. AB - OBJECTIVE: To examine the potential of surveys delivered via the World Wide Web (WWW) as a method for reaching hidden populations such as that of nonabusive users of illicit drugs. METHODS: Past and current approaches to collecting data from hidden populations were reviewed. RESULTS: A number of approaches have been used in the past efforts to reach drug users but the use of the WWW has produced the largest sample of successful illicit drug users ever surveyed. CONCLUSION: Further application of this approach (WWW) is recommended when hidden populations are surveyed. PMID- 12751618 TI - Stage of change specific triggers and barriers to moderate physical activity. AB - OBJECTIVE: To identify the proportion of rural Midwesterners at different stages of change for regular physical activity. METHODS: Through systematic random sampling, 418 adult subjects responded to telephone interviews about physical activity in 2 contiguous rural counties. RESULTS: Almost half of the population was in the precontemplation stage-suggesting a population at great risk for diseases with physical inactivity as a risk factor. Further, barriers and triggers differed by stage of change and demographic. CONCLUSIONS: Specific groups in this population would benefit by a campaign to increase self-efficacy, the availability of time for activity, and the awareness of associated benefits. PMID- 12751619 TI - Anal intercourse and sexual risk factors among college women, 1993-2000. AB - OBJECTIVE: To determine trends and sexual risk behaviors associated with anal intercourse among college women over an 8-year period. METHODS: A sexual activity questionnaire was used to collect data from 813 students enrolled in a women's health course. RESULTS: Thirty-two percent of the women had engaged in anal intercourse, and this measure was consistent across time. Women who had engaged in anal intercourse were significantly younger at first intercourse and had a greater lifetime number of partners and more reported STIs. CONCLUSION: This study underscores the importance of expanding our understanding of sexual behaviors of college women and openly addressing anal intercourse as a part of the sexual repertoire of college women. PMID- 12751620 TI - Diversifying the health professions: a model program. AB - OBJECTIVE: To describe a university-based mentoring program in the food and nutritional sciences that addresses the need for multicultural professionals in allied health fields. METHODS: The conceptual model for the program includes inputs (goals, resources), transformational process (professional development, social support and recognition) and outcomes (student participation, graduation and placement rates). Outcome data were analyzed using descriptive statistics. RESULTS: Between 1993 and 2001, 49 students completed the program, 43 (88%) graduated, 23 were placed in graduate or professional schools, and 21 obtained professional positions. CONCLUSION: This mentoring model resulted in multicultural students who successfully entered health professions. PMID- 12751621 TI - Do adults change their lifestyle behaviors after a cancer diagnosis? AB - OBJECTIVE: To examine changes in lifestyle behaviors after a cancer diagnosis and medical and demographic influences on such changes. METHODS: Adult cancer survivors (n = 352) completed a survey including demographic, medical, and lifestyle behavior change questions. RESULTS: Results showed that since cancer diagnosis, 46% of smokers quit smoking, 47% improved their dietary habits, and 30.1% exercised less. Adult cancer survivors who changed their lifestyle behaviors varied, depending on various demographic and medical variables and physician recommendation. CONCLUSION: It appears from our data that cancer diagnosis in adults may have a positive influence on smoking and diet and a negative influence on exercise. PMID- 12751622 TI - Social network positions and smoking experimentation among Chinese adolescents. AB - OBJECTIVE: To explore the relationship between peer social network positions and smoking experimentation among Chinese adolescents. METHODS: Self-administered questionnaires were administered to 1040 adolescents in grades 6, 8, and 10. Paired-friendship linkages were used to assign participants into 3 mutually exclusive social network positions. RESULTS: Overall isolates were more likely to have experimented with cigarettes than were group members or liaisons. However, among male 10th graders, more group members or liaisons had smoked than isolates. CONCLUSIONS: Smoking experimentation differs by social position, particularly among older adolescents. The association of social position with smoking experimentation could not be explained completely by traditional peer-related variables. PMID- 12751623 TI - Women's aging benchmarks in relation to their health habits and concerns. AB - OBJECTIVES: To identify aging benchmarks for women and to examine relationships between women's perceptions of aging and their health behaviors and concerns. METHODS: More than 1000 primarily white women completed paper-and-pencil and Internet surveys of demographic, attitudinal, and health-behavior information. RESULTS: Aging benchmarks correlated significantly with women's education and employment levels, reported health status, and health behaviors including exercise, dieting, and breast augmentation surgery. Those who worried about aging were significantly more likely to diet but not more likely to exercise. CONCLUSION: More research is needed to understand how perceptions influence women's ability to impact the aging process. PMID- 12751625 TI - Evaluation of folate conjugate uptake and transport by the choroid plexus of mice. AB - PURPOSE: Because the choroid plexus (CP) is enriched in cell surface folate receptors, an investigation was initiated to evaluate whether folate receptor mediated transcytosis might be exploited to deliver folate conjugates into the brain. METHODS: Balb/c mice were injected with radioactive and fluorescent conjugates of folate to measure and image their uptake by the CP. RESULTS: Retention of a radioactive folate conjugate, folate-diethylenetriaminepentaacetic acid (DTPA)-111In, into the brain of balb/c mice was observed, although repeated injections or prolonged release via an osmotic pump of the compound did not result in increased brain uptake. Uptake of an 125I-labeled anti-folate receptor antibody into the brain was very low, and no competition was observed with unlabeled antibody. Imaging of brain thin-sections and whole brain tissue from a mouse injected with folate-fluorescein revealed strong fluorescence in the CP, but virtually no where else in the brain. CONCLUSIONS: Both fluorescence and radioimaging results demonstrate specific uptake of small molecular weight folate conjugates into CP cells of the murine brain, but no significant transport of the molecules across the CSF. Furthermore, no uptake of larger folate-linked proteins by choroid plexus cells is observed, suggesting folate conjugate size may strongly influence access to CP folate receptors. PMID- 12751624 TI - In vivo and in vitro assessment of baseline blood-brain barrier parameters in the presence of novel nanoparticles. AB - PURPOSE: Nanoparticles have advantage as CNS drug delivery vehicles given they disguise drug permeation limiting characteristics. Conflicting toxicological data, however, is published with regard to blood-brain barrier integrity and gross mortality. METHODS: To address this issue two novel nanoparticle types: "emulsifying wax/Brij 78" and "Brij 72/Tween 80 nanoparticles were evaluated in vivo for effect on cerebral perfusion flow, barrier integrity, and permeability using the in situ brain perfusion technique. Additional evaluation was completed in vitro using bovine brain microvessel endothelial cells for effect on integrity, permeability, cationic transport interactions, and tight junction protein expression. RESULTS: In the presence of either nanoparticle formulation, no overall significant differences were observed for cerebral perfusion flow in vivo. Furthermore, observed in vitro and in vivo data showed no statistical changes in barrier integrity, membrane permeability, or facilitated choline transport. Western blot analyses of occludin and claudin-1 confirmed no protein expression changes with incubation of either nanoparticle. CONCLUSIONS: The nanoparticle formulations appear to have no effect on primary BBB parameters in established in vitro and in vivo blood-brain barrier models. PMID- 12751626 TI - Expression and functional involvement of organic anion transporting polypeptide subtype 3 (Slc21a7) in rat choroid plexus. AB - PURPOSE: It has been shown that transport(s) are involved in the uptake of estradiol 17beta glucuronide (E217betaG) by the choroid plexus (CP). The purpose of this study is to compare the substrate specificity of the transporter in the CP with those of rat organic anion transporting polypeptide 1 (rOatp1) and rOatp3. METHODS: The expression of rOatp1 and rOatp3 in rat CP was confirmed by RT-PCR and Western blot analyses. The substrate specificity of rOatp1 and rOatp3 was compared using cDNA-transfected LLC-PK1 cells. The uptake of E217betaG by rat isolated CP was determined by centrifugal filtration technique. RESULTS: PCR analyses demonstrated that the mRNA expression of rOatp3 was abundant in the CP, whereas that of rOatp1 was low. Immunohistochemical staining revealed that rOatp3 is expressed on the apical membrane of the CP. Kinetic parameters (Km and Ki values) of rOatp3 were similar to those for rOatp1. The results of mutual inhibition study suggest that E217betaG and taurocholate share the same mechanism in the CP. Corticosterone, estrone-3-sulfate and indomethacin are moderate inhibitors, but no effects by digoxin, p-aminohippurate, benzylpenicillin and cimetidine were observed. CONCLUSIONS: rOatp3 is most possible candidate transporter involved in the uptake of organic anions on the brush border membrane of the choroid epithelial cells. PMID- 12751628 TI - In vitro gene transfection in human glioma cells using a novel and less cytotoxic artificial lipoprotein delivery system. AB - PURPOSE: To develop and evaluate a novel artificial lipoprotein delivery system for in vitro gene transfection in human glioma cells. METHOD: Nanoemulsion was formulated with similar lipid compositions present in natural lipoproteins. The oil phase of nanoemulsion was composed of triolein (70%), egg phosphatidylcholine (22.7%), lysophosphatidylcholine (2.3%), cholesterol oleate (3.0%), and cholesterol (2.0%). To replace the surface protein as in natural lipoprotein, poly-L-lysine was modified to add palmitoyl chains at a basic condition and was incorporated onto the nanoemulsion particles through hydrophobic interaction. A model plasmid DNA, pSV-beta-Gal containing a reporter gene for beta-galactosidase was carried by the nanoemulsion/poly-L-lysine particles. The charge variation of soformed complex was examined by agarose gel electrophoresis and zeta potential measurement. In vitro transfection was conducted on human SF-767 glioma cell line using this new system. After standard X-Gal staining, transfected cells were observed under light microscope. The effect of chloroquine on the transfection was examined and, finally, the cytotoxicity of this new system was evaluated in comparison with commercial Lipofectamine gene transfection system. RESULTS: The plasmid DNA was effectively carried by this artificial lipoprotein delivery system and the reporter gene was expressed in the glioma cells. Transfection efficiency was significantly increased by the treatment of chloroquine, indicating that endocytosis possibly was the major cellular uptake pathway. Compared to Lipofectamine system, this new delivery system demonstrated similar transfection efficiency but a much lower cytotoxicity. In the experiment, the cell viability showed up to 75% using this system compared to only 24% using Lipofectamine system. CONCLUSION: A new artificial lipoprotein delivery system was developed for in vitro gene transfection in tumor cells. The new system showed similar transfection efficiency but a much lower cytotoxicity compared with commercial Lipofectamine system. PMID- 12751627 TI - Correlation of subcellular compartmentalization of HPMA copolymer-Mce6 conjugates with chemotherapeutic activity in human ovarian carcinoma cells. AB - PURPOSE: Intracellular targets sensitive to oxidized damage generated by photodynamic therapy (PDT) utilizing N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer-mesochlorin e6 monoethylenediamine (Mce6) conjugates was explored to aid in the design of second generation PDT delivery systems. METHODS: Low temperature, metabolic inhibitor, and nuclear localization sequences (NLS(FITC)) were used to achieve desired subcellular localization that was evaluated by confocal analysis and subcellular fractionation. Mce6 was bound to HPMA copolymer conjugates via non-degradable dipeptide linkers (P-GG-Mce6, P-NLS(FITC)-GG-Mce6) or lysosomally degradable tetrapeptide spacers (P-GFLG-Mce6, P-NLS(FITC)-GFLG Mce6). Chemotherapeutic efficacy was assessed by the concentration that inhibited growth by 50% (IC50), cell associated drug concentration (CAD) and confocal microscopy. RESULTS: P-GFLG-Mce6 possessed enhanced chemotherapeutic activ ity compared to P-GG-Mce6 indicating enzymatically released Mce6 was more active than copolymer-bound Mce6. Lysosomes appeared less sensitive to photodamage as observed by a higher IC50. Nuclear-directed HPMA copolymer-Mce6 conjugates (P NLS(FITC)-GG-Mce6, P-NLS(FITC)-GFLG-Mce6) possessed enhanced chemotherapeutic activity. However, control cationic HPMA copolymer-Mce6 conjugates containing a scrambled NLS (P-scNLS(FITC)-GG-Mce6) or amino groups (P-NH2-GG-Mce6) also displayed increased chemotherapeutic activity. CONCLUSIONS: Nuclear delivery was observed for P-NLS(FITC)-GG-Mce6 and P-NLS(FITC)-GFLG-Mce6 indicating NLS was a feasible approach for nuclear delivery. Due to the cationic nature of NLS, increased membrane binding of PDT systems incorporating cationic nuclear targeting moieties must be addressed. PMID- 12751629 TI - Poor correlation between intestinal and hepatic metabolic rates of CYP3A4 substrates in rats. AB - PURPOSE: To clarify the contribution of the intestinal first-pass metabolism to the drug bioavailability, the correlation between the intestinal and hepatic metabolism of human CYP3A4 substrates was investigated in rats. METHODS: The metabolic rates of four compounds (lidocaine, quinidine, nifedidpine, and rifabutin) were examined with excised intestinal tissues and liver microsomes. The intestinal and hepatic expression of CYP3A1/23 and CYP3A2 was evaluated by Western blot analysis. RESULTS: Rifabutin was metabolized fastest, and lidocaine was metabolized slowest in excised intestinal tissues. By contrast, lidocaine was metabolized fastest and rifabutin was the slowest in liver microsomes. The hepatic metabolism of lidocaine was inhibited by a CYP2D6 substrate desipramine, not by a CYP3A4 inhibitor ketoconazole. In addition, members of the CYP3A subfamily expressed in the intestine were different from those expressed in the liver. CONCLUSIONS: Poor correlation between the intestinal and hepatic metabolism of human CYP3A4 substrates in rats may be caused by the contribution of the CYP2D subfamily to the drug metabolisms in the liver and also by the unique expression of the CYP3A subfamily in the intestine. PMID- 12751630 TI - Human jejunal permeability of cyclosporin A: influence of surfactants on P glycoprotein efflux in Caco-2 cells. AB - PURPOSE: The purpose of this work was to determine the jejunal permeability of cyclosporin A (CsA) in humans and whether formulation variables modulate the effects of P-glycoprotein (P-gp) on the permeability of CsA in Caco-2 cells. METHODS: A solution containing CsA, phenylalanine, propranolol, polyethyleneglycol (PEG) 400, and PEG 4000 was perfused through a 10-cm jejunal segment in 12 subjects. Caco-2 transport studies were performed using previously reported methodology. RESULTS: The mean Peff (+/- SD) of CsA in humans was 1.65 (0.53). The mean permeabilities for phenylalanine, propranolol, and PEG 400 were 4.54 (2.39), 2.90 (1.28), and 0.83 (0.51) x 10(-4) cm/s, respectively. The presence of surfactants significantly decreased the permeabilities of CsA in both directions in Caco-2 cells. CONCLUSIONS: The results suggest that the effects of surfactants via micellar solubilization and inhibition of P-gp efflux on CsA transport in Caco-2 cells are significant. CsA can rightly be classified as a low solubility-high permeability Class II BCS drug and its highly variable absorption from Sandimmune oral formulations is the result of poor dissolution characteristics. PMID- 12751631 TI - Midazolam exhibits characteristics of a highly permeable P-glycoprotein substrate. AB - PURPOSE: The purpose of this study was to investigate whether midazolam exhibits characteristics of a highly permeable P-glycoprotein (P-gp) substrate and to evaluate the potential influence of P-gp inhibition on 1-OH midazolam formation during midazolam transport. METHODS: P-gp interaction was investigated by P-gp ATPase assay, efflux inhibition studies, and transport studies of midazolam across MDR1-MDCK and 1-alpha,25-dihydroxy vitamin D3-induced Caco-2 monolayers with and without the P-gp inhibitor GF120918. RESULTS: Midazolam was highly permeable and transport appeared essentially unpolarized. In MDR1-MDCK, the basolateral-to-apical (B-to-A) permeability was slightly higher (16%) than apical to-basolateral (A-to-B) permeability (p = 0.04); GF120918 increased A-to-B permeability by 27% (p = 0.01), and increased cellular midazolam accumulation during A-to-B transport by 45% (p = 0.01). Midazolam (200 microM) decreased rhodamine123 and vinblastine B/A ratios 3-fold (p < 0.006), while increasing their cellular accumulation (p < 0.003). P-gp ATPase activation by midazolam was dose-dependent and saturable [Km = 11.5(+/- 4.0) microM; Vmax = 41.1(+/- 7.4) nmol/mg/min]. P-gp inhibition increased 1-OH midazolam formation in A-to-B studies 1.3-fold when midazolam donor > or = 10 microM (p < 0.03). In B-to-A studies, P-gp inhibition did not significantly increase metabolite formation (p = 0.06). Midazolam's extraction ratio was not influenced by P-gp (p = 0.2). CONCLUSION: The results indicate that midazolam exhibited characteristics of a highly permeable P-gp substrate. 1-OH midazolam formation during A-to-B midazolam transport increased slightly when P-gp was inhibited. PMID- 12751632 TI - Differential effect of acute hepatic failure on in vivo and in vitro P glycoprotein functions in the intestine. AB - PURPOSE: The expression and function of P-glycoprotein (P-gp) in the intestine in carbon tetrachloride-induced acute hepatic failure (AHF) were evaluated in rats. METHODS: The expression of P-gp, in vivo absorption and exsorption of P-gp substrates (digoxin and rhodamine 123), and in vitro efflux transport of these P gp substrates were studied in the absence and presence of a P-gp inhibitor (verapamil or cyclosporin A) using the distal region of small intestine of control and AHF rats. RESULTS: Western blot analysis revealed that intestinal P gp expression level remained unchanged, or rather increased, in AHF. The in vivo intestinal P-gp function was significantly lower in AHF, as evaluated by the absorption and exsorption of P-gp substrates. In contrast, in vitro P-gp function was significantly higher in AHF, as evaluated by the efflux transport of P-gp substrates across the everted intestine. Collectively, the intestinal P-gp function was differently affected by AHF between in vivo and in vitro conditions. CONCLUSIONS: The in vivo intestinal P-gp function was suppressed in AHF, which could not be predicted from in vitro functional studies nor from P-gp expression level. The discrepancy between in vivo and in vitro results may be explained by the presence of endogenous P-gp inhibitors in the plasma of AHF rats. PMID- 12751633 TI - Lipophilicities of baclofen ester prodrugs correlate with affinities to the ATP dependent efflux pump P-glycoprotein: relevance for their permeation across the blood-brain barrier? AB - PURPOSE: Distribution to the effect site is a prerequisite for the therapeutic effect and determined by physicochemical properties and affinities to inside- and outside-directed membrane transporters. Based on the hypothesis that lipophilic esters of the GABA-derivative baclofen have a higher affinity to brain tissue, baclofen esters (methyl, ethyl, 1-propyl, 2-propyl, butyl) were studied regarding their penetration through the blood-brain barrier and their affinities to P glycoprotein (P-gp). METHODS: Octanol-water distribution coefficients (D) served as lipophilicity parameters. Blood and brain concentrations of baclofen and its methyl ester were determined in vivo in rats following intraperitoneal administration. Affinities to P-gp were evaluated using a radioligand binding assay based on P-gp-overexpressing cells and [3H]-talinolol as radioligand. RESULTS: Log D values for baclofen and ester derivatives were -0.96 (baclofen), 0.48 (methyl), 0.77 (ethyl), 1.31 (1-propyl), 1.27 (2-propyl), and 1.42 (butyl). In-vitro studies yielded negligible affinity of baclofen to P-gp, whereas IC50 values for the esters ranged between 1300 microM (methyl) and 290 microM (2 propyl). Affinity parameters correlated well with the lipophilicity parameters. CONCLUSION: Despite the P-gp affinity, brain concentrations of methyl ester were significantly higher than those of baclofen, however, baclofen levels following administration of the ester were smaller than with baclofen administration indicating only partial hydrolysis. PMID- 12751634 TI - Pathways and kinetics of deslorelin degradation in an airway epithelial cell line (Calu-1). AB - PURPOSE: The objective of this study is to investigate the pathways and kinetics of degradation of deslorelin, pGlu1-His2-Trp3-Ser4-Tyr5-D-Trp6-Leu7-Arg8-ProNHEt9 (Des1-9), in a human airway epithelial cell line (Calu-1). METHODS: The degradation of deslorelin in membrane and cytosolic fractions of Calu-1 cells was studied at 37 degrees C up to 24 h. The degradation products were separated using HPLC and identified by amino acid analysis, sequencing, and mass spectrometry. The rate constants for deslorelin degradation and the formation of degradation products were determined by fitting the concentration vs. time data to pharmacokinetic models using WinNonlin. The effect of enzyme inhibitors, captopril, phosphoramidon, and disodium EDTA on deslorelin degradation was also assessed. RESULTS: Des1-3, Des4-9, and Des5-9 were the deslorelin fragments detected in the membrane fraction. Apart from these degradation products. Des5-7 was also detected in the cytosolic fraction. The deslorelin degradation was 8.5 times faster in the cytosolic fraction compared to the membrane fraction. The disappearance of deslorelin and the kinetics of degradation products could be explained by simple 2 compartment iv bolus model and 1 compartment absorption model, respectively. EDTA and captopril decreased deslorelin degradation in the membrane and cytosolic fractions. CONCLUSIONS: Deslorelin is initially cleaved at the 3-4 bond in the membrane and cytosolic fractions, possibly by a metalloendopeptidase and/or angiotensin converting enzyme, with the degradation being more rapid in the cytosol. PMID- 12751635 TI - Large porous particle impingement on lung epithelial cell monolayers--toward improved particle characterization in the lung. AB - PURPOSE: The ability to optimize new formulations for pulmonary delivery has been limited by inadequate in vitro models used to mimic conditions particles encounter in the lungs. The aim is to develop a physiologically-relevant model of the pulmonary epithelial barrier that would allow for quantitative characterization of therapeutic aerosols in vitro. METHODS: Calu-3 human bronchial epithelial cells were cultured on permeable filter inserts under air interfaced culture (AIC) and liquid-covered culture (LCC) conditions. Calu-3 cells grown under both conditions formed tight monolayers and appeared physiologically similar by SEM and immunocytochemical staining against cell junctional proteins and prosurfactant protein-C. RESULTS: Aerosolized large porous particles (LPP) deposited homogeneously and reproducibly on the cell surface and caused no apparent damage to cell monolayers by SEM and light microscopy. However, monolayers initially grown under LCC conditions showed a significant decrease in barrier properties within the first 90 min after impingement with microparticles, as determined by transepithelial electrical resistance (TEER) measurements and fluorescein-sodium transport. Conversely, AIC grown monolayers showed no significant change in barrier properties within the first 90 min following particle application. A dense mucus coating was found on AIC grown Calu-3 monolayers, but not on LCC grown monolayers, which may protect the cell surface during particle impinging. CONCLUSION: This in vitro model, based on AIC grown Calu-3 cells, should allow a more relevant and quantitative characterization of therapeutic aerosol particles intended for delivery to the tracheobronchial region of the lung or to the nasal passages. Such characterization is likely to be particularly important with therapeutic aerosol particles designed to provide sustained drug release in the lung. PMID- 12751636 TI - Dissolution behavior of a poorly water soluble compound in the presence of Tween 80. AB - PURPOSE: To investigate the mechanism by which Tween 80 impedes the dissolution of CI-1041, a poorly water-soluble compound in its free form. METHODS: Bulk powder and intrinsic dissolution (ID) of CI-1041 in 0.1 N HCl with various concentrations of Tween 80 were conducted. The residual solids of the dissolution experiments were characterized. The surface tension and the critical micellar concentration (CMC) of Tween 80 in 0.1 N HCl were determined. RESULTS: CI-1041 underwent solvent mediated conversion to its chloride salt (CS) in 0.1 N HCl. The coating of the CS on the surface of the CI-1041 pellet decreased the ID rate 20 to 30 fold. When the Tween 80 concentration in 0.1 N HCl was below 0.5 mg/ml, the CS formation rate increased with increasing Tween 80 concentration. Above 0.5 mg/ml of Tween 80 in 0.1 N HCl, opposite trend was observed. The change in trend at 0.5 mg/ml Tween 80 coincided approximately with the CMC of Tween 80 in 0.1 N HCl. CONCLUSIONS: The authors propose the following mechanism mediated by Tween 80. Below CMC, reduced surface tension caused by addition of Tween 80 increases the rate of nucleation of insoluble CS, causing the formation of CS on the surface of the CI-1041 free form. This, in turn, decreases the dissolution rate by decreasing the release of compound into solution. Above CMC, the effect of reduced surface tension on the CS nucleation and therefore its formation may be negated by other factors, such as an increase in viscosity or adsorption of surfactant on the crystal surface. PMID- 12751637 TI - Crystalline to amorphous transition of disodium hydrogen phosphate during primary drying. AB - PURPOSE: To monitor the phase transitions during freeze-drying of disodium hydrogen phosphate. METHODS: The variable temperature sample stage of the X-ray diffractometer (XRD) was attached to a vacuum pump, which enabled the entire freeze-drying process to be carried out in the sample chamber. The phase transitions during the freeze-drying cycle were monitored in real time by XRD. Aqueous buffer solution (containing disodium hydrogen phosphate and sodium dihydrogen phosphate) was cooled at 2 degrees C/min from room temperature to -70 degrees C. It was then heated to -25 degrees C and subjected to primary drying for 2 h at a chamber pressure of approximately 100 mTorr, followed by secondary drying at -10 degrees C. RESULTS: In the frozen solution, disodium hydrogen phosphate had crystallized as the dodecahydrate (Na2HPO4 x 12H2O) as was evident from its characteristic lines at approximately 5.37, 4.27, and 2.81 angstroms. Primary drying for 2 h resulted in ice sublimation, and the complete disappearance of the dodecahydrate peaks. CONCLUSION: The dehydration of the crystalline dodecahydrate resulted in an amorphous anhydrate. Thus the amorphous nature of the end product is a result of phase transitions during the process and do not reflect the solid-state of the ingredients during the entire process. PMID- 12751638 TI - Effect of geometric structure of flow promoting agents on the flow properties of pharmaceutical powder mixture. AB - PURPOSE: The object of this work was to investigate the mechanism of how the surface geometric structure of flow agents affects on the flowability of pharmaceutical powder mixtures. METHODS: Nonporous and porous silicas were mixed with directly compressible fillers as flow promoting agents. The geometric structure of flow agents was investigated by gas adsorption and laser diffraction analysis. Flowability was evaluated with Carr's index measurement. Adhesion force between fillers and flow agents was determined using atomic force microscopy. RESULTS: Flowability was improved with the addition of both nonporous and porous flow agents. In the case of nonporous flow agents, effect to promote flowability decreased with the increase of particle diameter, whereas porous flow agents highly improved flowability independent of particle diameter. Atomic force microscopy measurement found that the adhesion force between a porous agent and filler was smaller than that between a nonporous agent and filler. CONCLUSIONS: Enhancement of flowability varies depending on the geometric structure of flow agents. Porous flow agents improve flow properties more than nonporous agents, because porosity is highly contributed to reduction of adhesion force between particles. PMID- 12751639 TI - Preparation and characterization of nanofibers containing amorphous drug dispersions generated by electrostatic spinning. AB - PURPOSE: We assessed the application of water-soluble polymer-based nanofibers prepared by electrostatic spinning as a means of altering the dissolution rate of the poorly water-soluble drug, itraconazole. METHODS: Organic solvent-based solutions of itraconazole/HPMC mixtures were electrostatically spun at 16 and 24 kV. The formed nanofibers were collected as a non-woven fabric. The samples were analyzed by scanning electron microscopy. differential scanning calorimetry, and dissolution rate. RESULTS: Scanning electron microscopy showed fiber diameters of 1-4 microm and 300-500 nm depending on the applied voltage. Differential scanning calorimetry measurements found that the melting endotherm for itraconazole was not present, suggesting the formation of an amorphous solid dispersion or solution. Dissolution studies assessed several presentations including direct addition of the non-woven fabrics to the dissolution vessels, folding weighed samples of the materials into hard gelatin capsules and placing folded material into a sinker. Controls included a physical mixture as well as solvent cast and melt extruded samples. Electrospun samples dissolved completely over time with the rate of dissolution depending on the formulation presentation and drug to polymer ratio. The physical mixture did not appreciably dissolve in these conditions. CONCLUSIONS: The application of electrostatic spinning to pharmaceutical applications resulted in dosage forms with useful and controllable dissolution properties. PMID- 12751640 TI - N-terminal site-specific mono-PEGylation of epidermal growth factor. AB - PURPOSE: N-terminal site-specific mono-PEGylation of recombinant human epidermal growth factor (EGF) was accomplished using polyethyleneglycol (PEG) derivatives (Mw = 2000 and 5000) through a reactive terminal aldehyde group. METHODS: The site-specific PEG conjugation was conducted ata slightly acidic pH condition (pH 5.5). The mono-PEGylation was targeted to an alpha-amine group at the N-terminal end of EGF to minimize reduction of biologic activity. Tryptic digestion mapping and MALDI-TOF MS techniques were applied to show the occurrence of mono PEGylation at the N-terminus of EGF. RESULTS: The site-specific mono-PEGylated EGF, when compared with native EGF, fully retained its in vitro biologic activities such a cell proliferation and intracellular signal transduction. This revealed that although a synthetic polymer of a PEG was covalently conjugated to EGF, the internalized complex of PEGylated EGF-receptor within cells did not hamper the intracellular signal transduction events. The PEGylated EGF also exhibited a prolonged circulation in blood stream in vivo and markedly enhanced physical stability whe incubated with tissue homogenate. CONCLUSION: N-terminally mono-PEGylated EGF shows increased physical stability while retaining its biologic activity. PMID- 12751641 TI - Targeting doxorubicin to epidermal growth factor receptors by site-specific conjugation of C225 to poly(L-glutamic acid) through a polyethylene glycol spacer. AB - PURPOSE: Targeted delivery of chemotherapeutic agents through antibody-polymer conjugates has met with limited success. One of the limiting factors is the loss of antibody's binding affinity upon conjugation with polymeric carriers because of lack of control over the number and site of attachment. This study aims to synthesize monovalent polymeric immunoconjugates through site-specific conjugation and to evaluate the in vitro binding activities of the resulting construct. METHODS: Antibody C225 against epidermal growth factor receptors was coupled to the terminus of a doxorubicin-bound block copolymer, poly(L-glutamic acid)-co-polyethylene glycol (PG-PEG). Western blot analysis, confocal fluorescent microscopy, and cytotoxicity assay were performed to confirm the specific binding of C225-PEG-PG-Dox to EGFR. RESULTS: C225 was conjugated to PEG PG-doxorubicin conjugates by reacting sulfhydryl group introduced to C225 with vinylsulfone group introduced at the terminus of PEG-PG block copolymer. Polymeric immunoconjugate C225-PEG-PG-Dox, but not control (i.e., conjugate without antibody), selectively bound to human vulvar squamous carcinoma A431 cells that overexpress epidermal growth factor receptors. Receptor-mediated uptake of C225-PEG-PG-Dox occurred rapidly (within 5 min), whereas nonspecific uptake of PEG-PG-Dox required an extended period of time (24 h) to internalize. Binding of C225-PEG-PG-Dox to A431 cells could be blocked by pretreatment with C225 antibody. C225-PEG-PG-Dox was more potent than free doxrubicin in inhibiting the growth of A431 cells after a 6-h exposure period. CONCLUSION: Site-specific conjugation of a monoclonal antibody to the terminus of a polymeric carrier enhances receptor-mediated delivery of anticancer agents. PMID- 12751642 TI - Wheat germ agglutinin binds to the epidermal growth factor receptor of artificial Caco-2 membranes as detected by silver nanoparticle enhanced fluorescence. AB - PURPOSE: The purpose of this study was to identify one of the ligands that mediate carbohydrate-specific cytoadhesion and cytoinvasion of wheat germ agglutinin (WGA)-containing drug delivery systems. METHODS: The receptor-ligand studies were performed with isolated epidermal growth factor (EGF) receptors as well as biomimetic membranes prepared from Caco-2 and A-431 cells. The binding of fluorescent labeled WGA was detected by the silver nanoparticle enhanced fluorescence technique. RESULTS: The binding of WGA to isolated EGF receptors is saturable and the equilibrium is reached within 1 min. The interaction between WGA and isolated EGF receptors is fully inhibited by the complementary carbohydrate and at least 85% of WGA binding to artificial Caco-2 membranes is caused by protein-carbohydrate interactions involving the tetrasialo-binding motif. The integrity and the presence of EGF-receptors in artificial Caco-2 membranes as well as their WGA-binding capacity were confirmed by immunoblot detection. CONCLUSIONS: The glycosylated extracellular domain III of the EGF receptor is involved in the WGA-Caco-2 cell interaction. Accordingly, receptor mediated endocytosis is the basic mechanism for internalization of WGA. As the EGF receptor is overexpressed in a high number of tumors but also occurs in non malignant tissue at considerable density, WGA-mediated drug delivery opens exciting possibilities for specific binding and uptake of poorly absorbable drugs. PMID- 12751643 TI - Long-term effects of the Positive Action program. AB - OBJECTIVE: To report long-term effectiveness of the Positive Action program. METHODS: Used matched-schools design and school-level achievement and disciplinary data to evaluate program effects on student performance and behavior in elementary schools. RESULTS: Participation in the Positive Action program improved student behavior, school involvement, and academic achievement at all 3 levels of schools, with the results showing a clear dose-response relationship. CONCLUSION: Results provide clear evidence that a coherent, comprehensive, and integrated program can have enduring effects in multiple domains. PMID- 12751644 TI - The relation of community developmental attentiveness to adolescent health. AB - OBJECTIVE: To investigate the relation of student exposure to selected developmental assets with indicators of thriving, using more reliable measures than previously reported. METHODS: Self-report surveys were completed by a diverse sample of 5136 students in 6th-12th grades. Analyses of variance were used to examine the assets-thriving relation. RESULTS: A majority of students reported experiencing only 3 of 12 assets. The more assets students reported, the more they reported thriving in physical, psychological, social, and academic domains. CONCLUSIONS: Adults must become more intentional about building the relationships, opportunities, values, and skills young people need for optimally healthy development. PMID- 12751645 TI - Diffusion of a developmental asset-building initiative in public schools. AB - OBJECTIVE: To identify characteristics that distinguish between school personnel who do and do not adopt asset-building behaviors in the formative stage of a countywide initiative. METHODS: An anonymous questionnaire, assessing constructs from diffusion of innovation and social cognitive theories, was administered to 467 adult personnel in 3 school districts. RESULTS: Discriminant analysis revealed that implementation of asset building was most closely related to rating the initiative high on trialability, relative advantage, compatibility, and observability and low on complexity. CONCLUSIONS: Efforts to implement asset building initiatives need to assess perceptions of adult personnel and should not assume that positive youth development will be widely supported in schools. PMID- 12751646 TI - Chicano-Latino Youth Leadership Institute: an asset-based program for youth. AB - OBJECTIVE: To describe the theory of action that links the Chicano-Latino Youth Leadership Institute with positive youth development and to highlight promising practices. METHODS: A logic model provided the basis for data collection through written surveys, focus groups, reflection forms, and analysis of Minnesota Student Survey Results. RESULTS: Primary participant outcomes include increased levels of community service, improved peer relationships, and higher graduation rates than those of Chicano-Latino non-participants. CONCLUSION: Based on the first 3 years of program evaluation data, this culturally specific youth development model is positively impacting a significant number of participants. PMID- 12751647 TI - Family involvement, problem and prosocial behavior outcomes of Latino youth. AB - OBJECTIVE: To explore the relations between familial factors and behavioral outcomes in Latino adolescents using a youth development framework. METHODS: A convenience sample of Latino youth (n = 446) completed an anonymous survey. Multiple linear regression was used to examine the associations between levels of family involvement and problem and prosocial behavior outcomes. RESULTS: Higher levels of parental monitoring and familial connectedness were consistently associated with less problem-behavior involvement. Sociocultural encouragement was consistently associated with more prosocial-behavior involvement. CONCLUSION: The family context is critical to the prevention of problem behaviors and the promotion of prosocial behaviors among Latino adolescents. PMID- 12751648 TI - Empowerment as fostering positive youth development and citizenship. AB - OBJECTIVE: To develop a theoretical framework of youth empowerment in the context of a participatory community health promotion intervention, a longitudinal qualitative study was conducted. METHODS: Individual and group interviews, documents, and observations were analyzed using the constant comparative method and theoretical sampling. RESULTS: Practitioners created an environment conducive to adolescents' taking responsibility for their quality-of-life issues by welcoming and enabling youth. Power was transferred to youth as responsibility for voicing, decision making, and action. This led to positive changes in youth development and their social integration into community. CONCLUSION: Empowerment emerged as a transactional partnering process between adults and youth. PMID- 12751649 TI - A youth development approach to profiling sexual abstinence. AB - OBJECTIVE: To use youth and parent demographic factors, youth assets (strengths), and community constructs to develop age-group-specific profiles for youth who abstain from sexual activity. METHODS: Data were collected using in-person interviews of youth (N = 1253) residing in randomly selected, inner-city neighborhood households. Data were analyzed to create classification trees. RESULTS: Results indicated that assets typically were the most important factors related to abstinence and that the profiles differed for each age group. CONCLUSIONS: When sexual abstinence is considered, the influence of youth and parent demographics, youth assets, and community factors appears to vary across adolescent developmental age periods. PMID- 12751650 TI - Treatment of viral conjunctivitis in children. PMID- 12751651 TI - Is tacrolimus the best therapy for atopic dermatitis? PMID- 12751652 TI - Importance of bioterrorism preparedness for family physicians. PMID- 12751653 TI - Update on the prevention and treatment of sexually transmitted diseases. AB - The Centers for Disease Control and Prevention (CDC) recently published updated guidelines that provide new strategies for the prevention and treatment of sexually transmitted diseases (STDs). Patient education is the first important step in reducing the number of persons who engage in risky sexual behaviors. Information on STD prevention should be individualized on the basis of the patient's stage of development and understanding of sexual issues. Other preventive strategies include administering the hepatitis B vaccine series to unimmunized patients who present for STD evaluation and administering hepatitis A vaccine to illegal drug users and men who have sex with men. The CDC recommends against using any form of nonoxynol 9 for STD prevention. New treatment strategies include avoiding the use of quinolone therapy in patients who contract gonorrhea in California or Hawaii. Testing for cure is not necessary if chlamydial infection is treated with a first-line antibiotic (azithromycin or doxycycline). However, all women should be retested three to four months after treatment for chlamydial infection, because of the high incidence of reinfection. Testing for herpes simplex virus serotype is advised in patients with genital infection, because recurrent infection is less likely with the type 1 serotype than with the type 2 serotype. The CDC guidelines also include new information on the treatment of diseases characterized by vaginal discharge. PMID- 12751654 TI - Recognition and management of bioterrorism infections. AB - Recent events have demonstrated that bioterrorists have the ability to disseminate biologic agents in the United States and cause widespread social panic. Family physicians would play a key role in the initial recognition of a potential bioterrorism attack. Familiarity with the infectious agents of highest priority can expedite diagnosis and initial management, and lead to a successful public health response to such an attack. High-priority infectious agents include anthrax, smallpox, plague, tularemia, botulism, and viral hemorrhagic fever. Anthrax and smallpox must be distinguished from such common infections as influenza and varicella. Anthrax treatment is stratified into postexposure prophylaxis and treatment of confirmed cutaneous, intestinal, or inhalation anthrax. Disease prevention by vaccination and isolation of affected persons is key in preventing widespread smallpox infection. Many resources are available to physicians when a bioterrorism attack is suspected, including local public health agencies and the Centers for Disease Control and Prevention. PMID- 12751656 TI - Information from your family doctor, What should I know about open-angle glaucoma? PMID- 12751655 TI - Open-angle glaucoma. AB - Glaucoma is the second most common cause of legal blindness in the United States. Open-angle glaucoma is an asymptomatic, progressive optic neuropathy characterized by enlarging optic disc cupping and visual field loss. Patients at increased risk for open-angle glaucoma include blacks older than 40 years, whites older than 65 years, and persons with a family history of glaucoma or a personal history of diabetes or severe myopia. Elevated intraocular pressure is a strong, modifiable risk factor for open-angle glaucoma, but it is not diagnostic. Some patients with glaucoma have normal intraocular pressure (i.e., normal-pressure glaucoma), and many patients with elevated intraocular pressure do not have glaucoma (i.e., glaucoma suspects). Routine measurement of intraocular pressure by primary care physicians to screen patients for glaucoma is not recommended. Open-angle glaucoma usually is discovered during an adult eye evaluation performed for other indications. Final diagnosis and treatment occur in collaboration with ophthalmologists and optometrists. Formal visual field testing (perimetry) is a mainstay of glaucoma diagnosis and management. Eye drops, commonly nonspecific beta-blocker or prostaglandin analog drops, generally are the first-line treatment to reduce intraocular pressure. Laser treatment and surgery usually are reserved for patients in whom medical treatment has failed. Without treatment, open-angle glaucoma can end in irreversible vision loss. PMID- 12751657 TI - Diagnosis and management of positional head deformity. AB - In children with positional head deformity (posterior plagiocephaly), the occiput is flattened with corresponding facial asymmetry. The incidence of positional head deformity increased dramatically between 1992 and 1999, and now occurs in one of every 60 live births. One proposed cause of the increased incidence of positional head deformity is the initiative to place infants on their backs during sleep to prevent sudden infant death syndrome. With early detection and intervention, most positional head deformities can be treated conservatively with physical therapy or a head orthosis ("helmet"). PMID- 12751658 TI - A practical approach to hypercalcemia. AB - Hypercalcemia is a disorder commonly encountered by primary care physicians. The diagnosis often is made incidentally in asymptomatic patients. Clinical manifestations affect the neuromuscular, gastrointestinal, renal, skeletal, and cardiovascular systems. The most common causes of hypercalcemia are primary hyperparathyroidism and malignancy. Some other important causes of hypercalcemia are medications and familial hypocalciuric hypercalcemia. An initial diagnostic work-up should include measurement of intact parathyroid hormone, and any medications that are likely to be causative should be discontinued. Parathyroid hormone is suppressed in malignancy-associated hypercalcemia and elevated in primary hyperparathyroidism. It is essential to exclude other causes before considering parathyroid surgery, and patients should be referred for parathyroidectomy only if they meet certain criteria. Many patients with primary hyperparathyroidism have a benign course and do not need surgery. Hypercalcemic crisis is a life-threatening emergency. Aggressive intravenous rehydration is the mainstay of management in severe hypercalcemia, and antiresorptive agents, such as calcitonin and bisphosphonates, frequently can alleviate the clinical manifestations of hypercalcemic disorders. PMID- 12751659 TI - Attention-deficit/hyperactivity disorder. PMID- 12751660 TI - Behavioral counseling in primary care to promote physical activity. PMID- 12751661 TI - Photo quiz. Infected welding burns. PMID- 12751666 TI - ACS updates guideline for the early detection of cervical neoplasia and cancer. American Cancer Society. PMID- 12751667 TI - Do I have to resuscitate this patient against her wishes? PMID- 12751668 TI - Emerging insights into peripartum cardiomyopathy. AB - Peripartum cardiomyopathy (PPCM) is relatively common in the Hospital Albert Schweitzer (HAS) district of Haiti. This investigation was carried out to expand epidemiologic data aiming at identifying risk factors for PPCM in this population. The HAS District PPCM Registry with 74 PPCM patients, enrolled from 1 February 2000 to 1 September 2002, served to identify the PPCM patients involved in this study. Thirty-seven non-PPCM Haitian mothers from the HAS district served as controls in the case-control study I and 32 non-PPCM Haitian mothers from the HAS district served as controls for the case-control study II. Following informed consent, patients and controls participated in clinical examination, echocardiography, epidemiologic questionnaire interviews, and immunohaematologic testing. Findings revealed: increased parity in PPCM vs control mothers (4.6 vs 3.3, p = 0.0252); 47% of the PPCM mothers had their initial diagnosis with the 5th or more pregnancy; increased number of patients with some hospital prenatal care in PPCM vs control mothers (42% vs 0%, p = 0.00001); and increased valley unit PPCM vs control mothers with no formal schooling (54% vs 24%, p = 0.0054). However, when hill-unit controls were included, there was no statistical difference in this category. Taking drinking-water from the river was found in 11% of the valley PPCM mothers vs 0% of the valley control mothers (p = 0.0509). Although the first 59 PPCM cases identified came from the valley units, recent identification of PPCM mothers in the hill unit indicates similar incidence of PPCM in mothers of remote hill area compared to the valley mothers--approximately 1 per 350 to 400 livebirths. Although the cause of PPCM and reasons for increased incidence in the HAS district of Haiti remain unknown, initial data present emerging insights and avenues to pursue in subsequent studies. PMID- 12751669 TI - Perinatal mortality in northern rural Tanzania. AB - The study was conducted to investigate the association between perinatal mortalityand factors relating to nutrition and infections in a rural population in northern Tanzania. A cohort of 3,618 women attending antenatal clinics was registered with background information and the results of antenatal examinations, and followed up after delivery. Stillbirths and neonatal deaths were identified and traced for an interview with the closest relatives. No information on outcome of pregnancy was obtained for seven women, and incomplete information was obtained for 99. The perinatal mortality rate was 27/1,000 births [95% confidence interval (CI) 22/1,000-33/1,000]; 44% were early neonatal deaths; and 56% were stillborn. There was an increased risk of perinatal death among babies with low birth-weight [for babies weighing 2,000-2,499 g, adjusted odds ratio (AOR) 5.8, 95% CI 2.1-15.8, babies below 2,000 g AOR 45.7; 95% CI 18.3-114.1], babies of women with a small arm circumference (below 23 cm, AOR 5.3, 95% CI 1.3-22.2), babies of women with positive VDRL serology (AOR 5.1, 95% CI 1.0-25.7), babies of mothers who had previously lost a baby (AOR 1.9, 95% CI 1.1-3.2), and among babies of nulliparous women (AOR 1.7; 95% CI 1.0-3.0). Infections and nutritional deficiencies should be addressed at antenatal clinics. PMID- 12751670 TI - Plasma vitamin A and C status of in-school adolescents and associated factors in Enugu State, Nigeria. AB - The study was undertaken to determine the intake of vitamin A and C and plasma concentrations of these vitamins among in-school adolescents. The factors affecting the vitamin status of these adolescents were also determined. Data for this report were obtained from a cross-sectional survey of 600 in-school adolescents in Nsukka local government area (LGA) of Enugu State, Nigeria. Ninety and 45 of the adolescents were randomly selected for determining plasma concentrations of vitamin A and C and intake of these vitamins respectively. Dietary assessment was done using a three-day weighed food intake method. Venous blood samples were collected and used for determining plasma vitamin A using the trifluoroacetic acid (TFA) method, while plasma vitamin C was determined by the thiourea method. Values obtained were matched against standards. The intake of vitamin A by all the adolescents was adequate (126-137% of recommended intake), while the intake of vitamin C was inadequate (51-91% of recommended intake). The mean intake of vitamin C was higher among males (23.7 +/- 0.71-27.3 +/- 8.0) than among females (15.3 +/- 2.8-19.5 +/- 5.1). Despite the adequate intake of vitamin A, 40% of the male and 32% of the female adolescents had low plasma concentrations of the vitamin (< 20 microg/dL). On the other hand, concentrations of plasma vitamin C were low among about 47% of these adolescents. Using multiple regression analysis, the two most important variables influencing vitamin A status were household size (b = -0.629; p < 0.0 1) and nutrition knowledge (b = 1.372; p < 0.01), while for vitamin C status, these were household size (b = 0.110; p = 0.05) and age (b = 0.226; p < 0.05). The daytime students had a significantly (p < 0.05) better vitamin A and C status than the boarders. The prevalence of vitamin A and C deficiencies among the adolescents may be more than estimated due to inadequate intake and/or poor bioavailability. This may pose a serious health risk for adolescents. There is, therefore, a need for adequate nutrition education and awareness about healthy lifestyles among Nigerian adolescents. PMID- 12751671 TI - Effect of chlorination of drinking-water on water quality and childhood diarrhoea in a village in Pakistan. AB - To evaluate the importance of public-domain transmission of pathogens in drinking water, an intervention study was carried out by chlorinating the public water supply system in a village in Pakistan. The water quality improved and reached a geometric mean of 3 Escherichia coli per 100 mL at the last standpipe of the water-supply system. Drinking-water source used and the occurrence of diarrhoea were monitored on a weekly basis over a six-month period among 144 children aged less than five years in the village. In this group, the children using chlorinated water from the water-supply scheme had a higher risk of diarrhoea than children using groundwater sources, controlled for confounding by season and availability of a toilet and a water-storage facility. The incidence of diarrhoea in the village (7.3 episodes per 10(3) person-days) was not statistically different from that in a neighbouring village where most children used water from a non-chlorinated water-supply system with very poor water quality. In this study area, under non-epidemic conditions, the reduction of faecal bacteria in the public drinking-water supply by chlorination does not seem to be a priority intervention to reduce childhood diarrhoea. However, the study was of limited size and cannot provide conclusive evidence. PMID- 12751672 TI - Aetiology of shigellosis in northern Pakistan. AB - People of northern Pakistan face health hazards because of poor sanitation practices. Bacterial gastrointestinal infections are very common, and sometimes outbreaks occur. The present study was aimed at evaluating and analyzing infestation of Shigella spp. in patients with suspected gastroenteritis and ascertaining the status of antibiotic therapy. Five hundred and eighty-five faecal samples of patients with suspected gastroenteritis, referred to the District Headquarter Hospital Gilgit, were investigated for common enteropathogenic bacteria from July 1997 to September 1999. Seventy-seven (13.2%) of the faecal specimens were infected with different strains of Shigella spp., 61% of which were Shigella dysenteriae, 15.6% were S. flexneri, and 23.4% were Shigella sp. All Shigella strains were sensitive to ceftriaxone, cefotaxime, ciprofloxacin, and enoxacin. Sixty-one percent of the strains were resistant to both ampicillin and chloramphenicol, and 3.9% to ampicillin and nalidixic acid, while 10.4% were resistant to ampicillin alone and 14.3% to chloramphenicol only. Only 10.4% of the strains were sensitive to all the antibiotics tested. Sixty strains of Shigella spp. were processed for isolation of plasmids, and 58 (97%) of these antibiotic-resistant bacteria harboured at least one plasmid. The number of plasmids varied from 1 to 9. Escherichia coli C600 were transformed with the isolated plasmids. Transformants, containing 23-kb plasmid, resisted growth in media containing antibiotics, thereby indicating that antibiotic resistance is plasmid-borne. Based on the findings of the study, it is concluded that the infestation of Shigella spp. is high in northern Pakistan, the aetiological agents are highly resistant to chloramphenicol and ampicillin, and the antibiotic resistance is mediated by the 23-kb plasmid. PMID- 12751673 TI - Factors associated with non-use of maternal health services in Botswana. AB - The study investigated individual and household factors associated with non-use of maternal health services in Botswana. Nationally-representative data, drawn from the 1996 Botswana Family Health Survey, were used. A weighted sample of 19,031 women, aged 15-49 years, who had at least one pregnancy history in the five years prior to the survey was considered for analysis. Both simple cross tabulations and logistic regression were used for analyzing the data. Consistently, the teenagers were less likely to seek prenatal care, to have their babies delivered by a qualified person, and to seek postnatal check-up. Using results from logistic regression analysis, it can be observed that low-parity women were less likely to use maternal services. Another consistent finding is that women with low educational level, those residing in rural areas, and those with low socioeconomic status were less likely to use maternal services. More focussed investigation is needed, but understanding the differentials of the use of maternal services allows policy-makers to identify problem areas that need attention. PMID- 12751674 TI - Nature of domestic violence against women in a rural area of Bangladesh: implication for preventive interventions. AB - This paper reports finding from a study carried out in a remote rural area of Bangladesh during December 2000. Nineteen key informants were interviewed for collecting data on domestic violence against women. Each key informant provided information about 10 closest neighbouring ever-married women covering a total of 190 women. The questionnaire included information about frequency of physical violence, verbal abuse, and other relevant information, including background characteristics of the women and their husbands. 50.5% of the women were reported to be battered by their husbands and 2.1% by other family members. Beating by the husband was negatively related with age of husband: the odds of beating among women with husbands aged less than 30 years were six times of those with husbands aged 50 years or more. Members of micro-credit societies also had higher odds of being beaten than non-members. The paper discusses the possibility of community centred interventions by raising awareness about the violation of human rights issues and other legal and psychological consequences to prevent domestic violence against women. PMID- 12751675 TI - Knowledge of, attitudes towards, and practices relating to child-spacing methods in northern Burkina Faso. AB - This study was carried out to document current trends in knowledge of, attitudes towards, and practices relating to traditional and modern child-spacing methods in a remote area in northern Burkina Faso. Information on sexual abstention, weaning, and contraception was elicited from 296 women of reproductive age, involving 413 postpartum intervals. A number of older women and key informants were also interviewed. The findings depicted significant diversity in that durations of individual postpartum sexual abstinence varied between 40 days and 3 years, with shorter durations associated with stricter adherence to Islamic beliefs and, possibly, a trend towards a less collective and, for the family unit, more labour intensive, agro-pastoral subsistence economy. Although durations of amenorrhoea were relatively short at between (median) 9 and 11 months, they determined the length of non-susceptible periods in almost 90% of cases. The median timing of weaning was stable at 24 months across all three main ethnic groups. However, changes in the frequency and type of complementary feeds may have impacted on the duration of amenorrhoea. Both demand for modem contraception and contraceptive prevalence (< 1%) were very low. The creation of new child-spacing norms and the promotion of modern contraceptive methods are likely to be successful in areas like this one only, if the population can be sensitized to the idea that Islam does not necessarily discourage contraception. PMID- 12751676 TI - Common transfusion-transmitted infectious agents among thalassaemic children in Bangladesh. AB - Transfusion-dependent children are more prone to acquiring various transfusion transmitted infections (TTIs), such as hepatitis B (HBV), hepatitis C (HCV), HIV, and others. Since the magnitude of these infections among thalassaemic children in Bangladesh is not well-known, this study was conducted to assess the prevalence of TTIs among them (who received more than three blood transfusions) compared to their age- and sex-matched controls (non-thalassaemics and those who had never had a transfusion). Seromarkers for HBV, HCV, HDV, Treponema pallidum, and HIV were tested, and the results were analyzed using SPSS/Windows 10.5. Of 259 children studied, 152 (58.69%) were thalassaemic (mean age 6.8 +/- 3.6 years), and 107 were controls (mean age 6.7 +/- 3.53 years). The HBV and HCV markers were found significantly more often among multi-transfused thalassaemic children than among the controls in terms of HBsAg (13.8% vs 6.5%, p < 0.04), anti-HBc total (39.5% vs 9.4%, p < 0.0001), and anti-HCV (12.5% vs 0.9%, p < 0.0001). HBeAg did not differ (p = 0.82) between the thalassaemics (9.52%) and the controls (14.28%), whereas anti-HBe differed (0% vs 57.14%, p < 0.003). Neither the thalassaemics nor the controls were positive for HDV, HIV, or T. pallidum. Since more thalassaemic children acquired hepatitis B and C infections through multiple blood transfusions, it is recommended that the safe blood transfusion programme be strengthened and mass vaccination against HBV (even who suffer from HCV) in Bangladesh be undertaken. PMID- 12751677 TI - Serum iron, zinc and copper levels and lipid peroxidation in children with chronic giardiasis. AB - This study investigated the levels of iron, zinc, and copper and their demolishing effects against lipid peroxidation in chronic giardiasis. Serum iron, zinc and copper levels, erythrocyte cytosolic superoxide dismutase activity, and malondialdehyde levels were measured in 34 children with chronic giardiasis and were compared with controls. The serum iron and zinc levels and erythrocyte superoxide dismutase activity were significantly lower, and malondialdehyde levels were significantly higher among the children with chronic giardiasis compared to the control group (p < 0.001). There was no significant difference in copper levels between the two groups (p > 0.05). Consequently, the oxidant antioxidant balance may tilt towards the oxidative side due to weakness of the antioxidant system in giardiasis. If early and proper treatment is not performed, free radical-mediated damage might occur in children with chronic giardiasis. PMID- 12751678 TI - Tetracycline in the treatment of cholera caused by Vibrio cholerae O1 resistant to the drug in vitro. PMID- 12751679 TI - Emergence of multidrug-resistant Shigella dysenteriae type 1 causing sporadic outbreak in and around Kolkata, India. PMID- 12751680 TI - Inherited autosomal dominant polycystic kidney disease--genetic polymorphism. PMID- 12751681 TI - Ten-year clinical and ultrasonographic follow-up of siblings from families with autosomal dominant polycystic kidney disease. AB - We conducted a ten-year clinical and ultrasound follow-up study of 120 siblings with ADPKD (68 men and 52 women, aged 19-40). 40 subjects had polycystic kidney disease. During the study period, the number and size of the cysts increased. Symptoms and signs also changed: at baseline 51% of the subjects were asymptomatic dropping subsequently to 2%. Initially, 32 subjects had 1-5 cysts in one or both kidneys and they were classified as suspected of having ADPKD. Significant changes were found in this group at the end of the follow-up. In 12 of them (37.50%) subsequent ultrasonograms revealed an increase in the number and size of the cysts--i.e. evolution towards ADPKD. None of the subjects in this group had a decrease in the number of cysts. In the control group, three had multiple cysts but most subjects were ultrasonographically negative for polycystic kidney disease. In conclusion, the authors recommend a clinical and ultrasonographic long-term follow-up of subjects at risk for ADPKD which should allow early diagnosis as well as prevention of the complications which result in chronic renal failure. PMID- 12751682 TI - Age-related changes of the somatosensory evoked potentials in healthy children. AB - AIM: The aim of the study was to investigate the configuration and latency of the somatosensory evoked potentials (SEPs) in healthy children for the time from the neonatal period to adolescence. MATERIAL AND METHODS: SEPs were recorded in 67 healthy children--37 boys and 30 girls from 0 to 16 years of age by means of median nerve stimulation. The active electrode was placed above the contralateral parietal cortex on places C3' and C4' and the reference electrode--on Fpz. The filters were 10-1000 Hz and the frequency of the electric stimulus--3 Hz. RESULTS: The depression of the potential in the neonatal period is accounted for by the wide base and low amplitude of SEPs. The configuration of SEPs is identical with that of adult individuals after 3 years of age. The latencies of the waves P15, N20 and P25 decrease progressively with age and lengthen in the period from 9 to 16 years of age, with the increase of height. The interpeak latencies P15 - N20, N20 - P25 and P15 - P25 decrease with age, while the amplitude N20/P25 increase with age. We found significant variability in the amplitude of the potential, most pronounced in the neonatal period. It gradually decreases by 10-14 months of age and after that remains unchanged. We did not find any differences at stimulation between the left and the right hand. CONCLUSION: The age-related changes in SEPs reflect the trends of the development and the maturation of the neural pathways and their better myelinization. PMID- 12751683 TI - Reproducibility and agreement between three methods of intraocular pressure measurement. AB - OBJECTIVE: The aim of the present study was to establish reproducibility between consecutive intraocular pressure measurements using the same method of tonometry as well as to assess agreement between 3 different methods of tonometry. MATERIALS AND METHODS: The study included 55 patients. Intraocular pressure of 94 eyes was measured, using Goldman applanation tonometry, Maklakoff applanation tonometry and indentation tonometry according to Schioetz method. The mean values and the differences in intraocular pressure measurements were calculated (using Bland and Altman method) in order to assess agreement between Goldman and Maklakoff applanation tonometry and Goldman and Schioetz tonometry. Reproducibility of the methods was assessed based on three consecutive intraocular pressure measurements on three groups of 20 eyes at 5-minute intervals. RESULTS: The mean intraocular pressure (Po) measured by Goldman method was 23.26 +/- 0.78 mmHg, the mean pressure estimates (Po) according to Maklakoff method were 19.4 +/- 0.64 mmHg and it (Po) was 21.18 +/- 0.74 mmHg using Schioetz tonometry. The correlation (Pearson) between Goldman Po and Maklakoff Po was 0.74 and between Goldman and Schioetz Po - 0.93. Mean differences were 4.12 +/- 0.53 and 2.08 +/- 0.3 mmHg respectively. Variation coefficients between consecutive measurements were: for Goldman tonometry - 2.00%, for Maklakoff - 7.90% and for Schioetz - 5.72%. CONCLUSIONS: According to Bland and Altman method there is low agreement between intraocular pressure estimates obtained by Goldman and Maklakoff tonometry, i.e. these methods are not interchangeable. This fact might be explained by the high variation coefficient of Maklakoff method. The low variation coefficient of Goldman tonometry makes it the most accurate of the three methods. PMID- 12751684 TI - Nonrheumatic atrial fibrillation and left ventricular hypertrophy in the prognosis of reversible ischaemic neurological deficit. AB - Nonrheumatic atrial fibrillation (NRAF) and left ventricular hypertrophy (LVH) have long been recognised as risk factors for cerebral ischaemia and as predictors of recurrent vascular events. In the present study we aimed at determining the value of NRAF and LVH as predictors of recurrent vascular events in a cohort of patients with a first-ever episode of reversible ischemic neurological deficit (RIND). The study included 54 patients (37 men and 17 women, aged 62 +/- 9.6 yrs) who had suffered RIND; they were followed up for 30 days after the stroke in clinical conditions and for 12 months as outpatients. The patients were studied during the hospital stay by means of routine tests (electrocardiography, standard laboratory tests) and specialised studies (computer tomography, echocardiography). By the end of the one-year outpatient follow up there were 8 (14.8%) recurrent cerebrovascular events. By combining the statistically significant cerebrovascular risk factors (male sex, sudden onset of the event and moderately high systolic and diastolic blood pressure) with factors not reaching statistical significance (LVH, NRAF) we developed a statistically significant prediction model for patients with RIND. PMID- 12751685 TI - Endodontic treatment of permanent teeth in children with a new calcium-carbamide formaldehyde paste. AB - AIM, MATERIAL AND METHODS: Endodontic treatment of a total of 303 teeth with pulpitis and periodontitis was performed with a new calcium-carbamide formaldehyde paste. In three children with fractured teeth with complications, the applied treatment was combined (2 teeth with apical osteotomy and 1 with cystectomy). Control examinations were carried out at months 6, 12 and 24 and clinically healthy teeth and teeth with complications were registered. RESULTS: In the pulpitis group the clinically healthy teeth were 97.55% +/- 1.08 at the 6th month; 96.85% +/- 1.26 at the 12th month and 92.92% +/- 2.41 at the 24th month. In teeth with periodontitis 91.56% +/- 3.05 were clinically healthy at the 6th month; 93.83% +/- 2.67 at the 12th month and 98.03% +/- 1.98 at the 24th month. No statistical significance of the differences between pulpites and periodontites was found (P > 0.05). Recovery of the bone structure was observed in the teeth with combined endodontic and surgical treatment at the 12th month. CONCLUSIONS: The calcium-carbamide-formaldehyde paste stimulates bone regeneration and closing of the apex without coloring the hard dental tissues. PMID- 12751686 TI - Anti-Candida antibodies in serum and saliva of patients with denture stomatitis. AB - OBJECTIVE: Quantitative measurement of IgA and IgG antibodies in the serum and saliva of patients with denture stomatitis (stomatitis subprothetica) was the object of this study. METHODS: The indirect immunofluorescence technique (IFA) utilizing CIP 628 (serotype A) Candida albicans blastospores was employed. Anti human goat and rabbit fluorescein isothiocyanate labeled serum was used. Serial serum dilutions from 1:20 to 1:1280 and saliva serial dilutions from 1:1 to 1:8 were prepared. Specimens were read using an ML-2 fluorescence microscope. Alternative analysis, analysis of variance and graphical analysis was used for statistical analysis of data. RESULTS: Anti-Candida antibodies (mainly IgG class) were detectable at dilutions of 1:40, 1:80 and 1:160 in the serum of patients with denture stomatitis compared with healthy controls. The saliva samples of these patients showed higher titers of anti-Candida antibodies compared with controls (1:8 for IgG and 1:4 for IgA). CONCLUSIONS: IgG anti-bodies were prevalent in the saliva of our patients and they were of diagnostic value. The IgA titers did not remain constant and could be a determinant of the disease course. Each patient showed an individual serologic profile. For this reason individualized interpretation of saliva antibodies as well as a comparison with the serum anti-Candida antibodies is recommended. PMID- 12751687 TI - Electron microscopic study of the differentiation of rat white subcutaneous adipocytes in situ. AB - In the present study we investigated the process of differentiation of rat subcutaneous white adipocytes in situ using electron microscopy. MATERIAL AND METHODS: Rat fetuses (Wistar rat) at 15 - 21 days of pregnancy were used. Sections of the subcutaneous tissue of a hind leg were prepared for examination with transmission electron microscopy by routine techniques. RESULTS: The electron microscopic study showed that the embryonal differentiation of rat subcutaneous white adipose cells in situ started on the 15 - 16th day of gestation and was expressed by lipid accumulation and concomitant changes in the cell organelles. We observed the following in the differentiating adipocytes: 1. abundant glycogen granules; 2. appearance and accumulation of lipid droplets with gradual formation of several lipid droplets or one large lipid drop which pushed the nucleus to the periphery and rounded the cell; 3. large elongated mitochondria with densely packed transverse cristae; disintegration of cristae, swelling and vacuolization occurring in some of the mitochondria; 4. well developed endoplasmic reticulum with elongated and dilated cisternae; the initial predominance of rough endoplasmatic reticulum being superseded by prevalence of smooth endoplasmatic reticulum. The ultrastructural analysis we did revealed the consecutive stages in the adipocyte differentiation and the formation of adipocyte phenotype during the embryonal development of the rat. PMID- 12751688 TI - Giant fibroadenoma of the breast. AB - A case of giant fibroadenoma of the breast (size 23x20x13 cm and weight 2680 grams) in a 43-year-old woman with short stature and gracile body build is presented. Pericanalicular fibroadenoma was detected histologically. One year after mastectomy the patient is in good health, with no recurrences. The case is discussed in the light of the diagnostic difficulties, which lead to radical mastectomy. Giant fibroadenomas have to be differentiated from phylloid cystosarcoma by the lack of leaf-like structures and stromal cell atypia and from the breast hamartoma and asymmetric breast hypertrophy in girls by the lack of mammary lobules. Giant fibroadenoma should take its due place in the diagnostic algorithm of the breast tumors. PMID- 12751689 TI - Plasma neutrophil lipocalin, elastase-alpha1-antitrypsin complex and neutrophil protease 4 in preterm infants with respiratory distress syndrome. AB - Respiratory distress syndrome (RDS) and chronic lung disease of prematurity (CLD) are associated with inflammation of the airways and interstitial tissue of the lung. It is hypothesized that RDS severity and the risk of developing CLD may be correlated with neutrophil gelatinase-associated lipocalin (NGAL), a marker of leucocyte activity, human elastase-alpha1-antitrypsin complex (HEAT) or free and complexed neutrophil protease 4 (NP4), markers of proteolytic enzyme secretion from granulocytes. Thirty-three preterm infants with RDS were enrolled in the study and plasma sampled between 3 and 14 days of life. NGAL, HEAT and NP4 concentrations varied widely in infants with RDS. Significant correlations between subsequent development of CLD and plasma concentrations of HEAT and NP4, respectively, were found on days 3-4 of life, p=0.006 and p=0.02, respectively. PMID- 12751690 TI - Effect of glucose/insulin infusion and magnesium supplementation on serum and muscle sodium and potassium and muscle [3H]ouabain binding capacity in Type 1 diabetes mellitus. AB - Glucose has recently been found to decrease muscle potassium content. The aim of this study was to evaluate the effect of the infusion of glucose and insulin and the effect of magnesium supplementation on serum and muscle sodium and potassium and muscle [3H]ouabain binding capacity in patients with Type 1 diabetes mellitus and in controls. Muscle potassium and sodium content, muscle [3H]ouabain binding capacity and serum potassium and sodium concentrations were determined in 10 patients with Type 1 diabetes mellitus and in 5 controls before and after an euglycaemic, hyperinsulinaemic clamp, and after an intravenous magnesium load test. Nine of the patients with Type 1 diabetes mellitus were restudied after 24 weeks of oral magnesium oxide supplementation. Basic serum and muscle sodium and potassium and muscle [3H]ouabain binding capacity did not differ between groups. The infusion of glucose and insulin reduced muscle potassium content, whereas muscle sodium content was unchanged. There were no differences between groups. Oral magnesium oxide supplementation increased muscle potassium content by 6%. Muscle [3H]ouabain binding capacity was unchanged. In patients with Type 1 diabetes mellitus, the intravenous infusion of magnesium increased serum potassium concentration before but not after oral magnesium oxide supplementation. In controls, the infusion of magnesium did not affect serum potassium concentration. It was found that intravenous infusion of glucose and insulin decreases muscle potassium content, probably by shifting potassium from the muscle cells to the splanchnic organs. Oral magnesium oxide supplementation increases muscle potassium content in patients with Type 1 diabetes mellitus. The increase in serum potassium concentration owing to the intravenous infusion of magnesium could be used in the evaluation of magnesium status in patients with Type 1 diabetes mellitus. This, however, requires further investigation. PMID- 12751691 TI - Glucose metabolism in relatives of type 1 diabetic patients with albuminuria. AB - OBJECTIVE: Diabetic nephropathy in type 1 diabetes is associated with familial aggregation of diabetes. In order to explore the mechanisms behind this association, we assessed glucose metabolism in glucose-intolerant relatives of type 1 diabetic patients with (ALB+) or without (ALB-) elevated urinary albumin excretion rate (UAER). METHODS: Glucose tolerance and insulin secretion were assessed using an oral glucose tolerance test (OGTT) and insulin sensitivity was measured with the short insulin tolerance test (ITT). RESULTS: One hundred and fourteen parents and siblings of 43 type 1 diabetic patients with ALB+ (UAER > or = 20 microg/min) were identified and 93 parents and siblings of 39 patients with ALB- (UAER < 20 microg/min). From this pool, a further selection was made of those (25 and 13 relatives of patients with ALB+ and ALB-, respectively) with mild abnormalities of glucose metabolism (fasting plasma glucose < 7.8 mmol/L; 2 h plasma glucose > or = 7.8 mmol/L in the OGTT). No difference in insulin sensitivity was discernible between the two groups of relatives (KITT 3.3 +/- 1.0 vs. 3.2 +/- 1.0%/min, p=NS). Although there were no significant differences in the incremental areas under glucose or insulin curves (AUC) between relatives of ALB+ and ALB- in the OGTT, the insulin secretory response to the rise in plasma glucose was impaired in relatives of patients with ALB+ (insulin AUC/glucose AUC: 7.1 [1.1-30.8] vs. 9.8 [3.6-52.2], p=0.039). CONCLUSIONS: Glucose-intolerant relatives of patients with elevated UAER seem to be characterized by impaired insulin secretion. Genetic or environmental factors related to impaired insulin secretion may be important in the development of diabetic nephropathy. PMID- 12751692 TI - Beta-2-glycoprotein I antibodies in patients with thrombosis. AB - The laboratory diagnosis of antiphospholipid antibody syndrome currently requires two consecutive positive results in either lupus anticoagulant or anticardiolipin antibody assays. Antibodies against beta-2-glycoprotein I (abeta2-GPI) are suggested as a new marker for the syndrome. The inclusion of abeta2-GPI in the official diagnostic criteria has so far been precluded owing to lack of an international standard and also technical difficulties. Samples from 5367 consecutive patients sent to a national reference laboratory mainly because of various thrombotic events were studied. An IgG abeta2-GPI ELISA assay was performed in addition to lupus anticoagulant (dRVVT and PTT-LA) and IgG anticardiolipin antibody determinations to evaluate patient groups in which the new assay might be of value. From a total of 90 patients, 2.2% of the samples were abeta2-GPI positive; 51 patients had abeta2-GPI as the only positive antiphospholipid antibody marker; 20 patients had had a venous thrombosis and 14 an arterial thrombosis, 4 had pregnancy complications and 2 had thrombocytopenia. Relatively young patients with cerebrovascular ischaemic events seemed especially to present sole abeta2-GPI positivity. The abeta2-GPI positivity remained fairly constant in the 23 patients from whom follow-up samples were taken. It is concluded that the IgG abeta2-GPI assay seems to be a potentially important additional diagnostic tool for the antiphospholipid antibody syndrome. PMID- 12751693 TI - Validation of albumin determined in urine with the HemoCue point-of-care analyser. AB - Albumin concentrations persistently between 20 and 200 mg/L in first morning spot urine or urine collected overnight, referred to as microalbuminuria, indicate incipient nephropathy in diabetes mellitus. This study validates a new point-of care device, the HemoCue Urine Albumin analyser, for handling, accuracy precision and predictive values (PV+/-) at 20 mg/L. Over a period of 2 months, 200 microalbuminuria samples were selected at the Department of Clinical Chemistry according to the results from the Integra 700 instrument (Roche, R) and analysed on the same day using the HemoCue analyser (HemoCue, H) and the Immage instrument (Beckman, B), in all cases closely following the manufacturers' instructions. Only 137 results were within the measuring range of H, 10-150 mg/L. Comparisons yielded regression lines H=1.06R-7.2 (r2=0.94), H=1.08B-3.1 (r2=0.94) and R=1.00B+4.3 (r2=0.99). Inter-assay (n=24) CV% at 12 mg/L was H=18.2, R=4.2 and B=2.9 and at 65 mg/L: H=6.1, R=1.8 and B=2.6. Intra-assay duplicate CV% for H at 21-40 mg/L was 13.2, at 41-80 mg/L 10.8 and at 81-150 mg/L 9.2. Intra-assay repeatability (n=8) CV% at 28 mg/L was 7.2-13.8, at 57 mg/L 6.4-8.4 and at 105 mg/L 4.3-7.1. External quality assurance urine albumin (B) was +5.7% cf. nephelometry and (R) +1.0% cf. turbidimetry (n=6) method-group means. PV+/- values were (H versus R) 0.98/0.37 and (H versus B) 0.95/0.65. HemoCue is easy to handle. Results below 20 mg/L need to be confirmed at the central laboratory to exclude microalbuminuria. Values above 20 mg/L can be used to follow microalbuminuria, as precision allows discerning steps of 10 mg/L. PMID- 12751694 TI - Haemostatic complications in haemodialysis patients: effect of type of vascular access and dialysis filter. AB - Impaired haemostasis is common in patients with end-stage renal disease, and may cause either thrombotic or bleeding complications. The purpose of this study was to assess whether plasma markers of coagulation activation in patients undergoing chronic haemodialysis (HD) can identify high-risk individuals, and to test the relevancy of type of vascular access or dialysis filter. We measured plasma levels of prothrombin fragment 1+2, fibrin D-dimers and tissue factor in 82 HD patients before and after dialysis. Clinical endpoints during the year following blood sampling were thrombosis in blood access, changes in blood access, other thromboembolic events, bleeding complications, ischaemic vascular disease, or death. We found elevated baseline levels of all three parameters in HD patients, compared to normal reference ranges. Plasma levels of all parameters (particularly fibrin D-dimers) were significantly higher in patients with prosthetic grafts and central venous dialysis catheters than in patients with native vessels. Patients with AV-fistulas or grafts who had bleeding complications (n=7) had significantly higher plasma levels of fibrin D-dimer and prothrombin fragment 1+2. Bleeding complications also occurred more frequently among the patients with prosthetic grafts (3/18) or central venous dialysis catheters (3/11) compared with those with grafts from native vessels (1/53). Other than a bleeding tendency, our data do no show any correlation between coagulation parameters and other clinical complications during haemodialysis. In conclusion, we found elevated plasma levels of markers of coagulation activation among HD patients. High levels of D-dimers and prothrombin fragment 1+2 correlated to bleeding diathesis instead of thromboembolism, and this tendency was most pronounced in patients with prosthetic grafts. PMID- 12751695 TI - Bread wheat gliadin cytotoxicity: a new three-dimensional cell model. AB - BACKGROUND: In an attempt to clarify the role of gliadin toxicity in the pathogenesis of gluten intolerance (celiac disease), previous in vitro studies have been based on two-dimensional human cell cultures. However, the specific morphological and biochemical properties of in vivo tissue are better maintained in three-dimensional cell cultures (multicellular spheroids, MCS). The aim of this study was to develop a three-dimensional in vitro model to investigate the effects of gliadin on epithelial cells and broaden our understanding of the early tissue damage occurring in celiac disease. METHODS: The three-dimensionally growing Lovo cell line was exposed to increasing concentrations of peptic-tryptic digested bread wheat gliadin (from 125 to 1000 microg/mL) for 7 days in order to evaluate cell viability (colony-forming assay), and at the standard concentration of 500 microg/mL for 7 days in order to evaluate MCS diameters, volumes and cell morphology using light and electron microscopy. RESULTS: In comparison with the controls, the cell viability of the gliadin-treated MCS was significantly reduced (20-80%), but there was no difference in size. Various degrees of cell damage (autophagic vacuoles and intra-cytoplasmic lipid-like droplets) were detected by both light and electron microscopy. CONCLUSION: This is the first study investigating the effects of gliadin on MCS. Lovo MCS seem to be responsive to gliadin exposure, thus confirming previous results obtained using two-dimensional cell cultures. The data suggest that three-dimensional cell cultures may be useful in broadening our understanding of some of the early effects of gliadin peptides on epithelial cells. PMID- 12751696 TI - Determination of urinary free cortisol by liquid chromatography-tandem mass spectrometry. AB - Measurement of urinary free cortisol is clinically important in the diagnosis of Cushing's syndrome. While liquid chromatography (LC) with UV detection provides much better specificity than immunologic methods, certain drugs cause interference. Detection by mass spectrometry (MS) is a potentially superior method. Our analysis utilizes 1 mL urine spiked with 6-alpha-methylprednisolone as internal standard. The samples were extracted with dichlormethane and the extract was washed, evaporated to dryness and analyzed by LC-MS/MS operating in the negative mode after separation on a reversed-phase C18 column. The calibration curves for analysis of urinary cortisol exhibited consistent linearity and reproducibility in the range of 10-400 nmol/L. Inter-assay CVs were 4.0-7.6%, at mean concentrations of 21-153 nmol/L. The detection limit was 1 nmol/L (signal-to-noise ratio=3). The mean recovery of cortisol added to urine ranged from 67% to 87% and that of the internal standard from 71% to 76%. The regression equation for the LC-MS/MS (x) and HPLC (y) methods was: y=1.095x+8.0 (r=0.996; n=111). Drugs known to interfere with UV detection did not cause problems here. The sensitivity and specificity of the MS/MS method for urinary free cortisol offer advantages over HPLC with UV detection by eliminating drug interference. The higher equipment costs in comparison with HPLC methods using UV detection are balanced by higher throughput, thanks to shorter chromatographic run times. PMID- 12751697 TI - Activity of antioxidant enzymes in children from families at high risk of premature coronary heart disease. AB - A positive family history of coronary heart disease (CHD) is one of the most predictive risk factors of CHD. Many children with increased risk of CHD because of their positive family history of CHD do not present other risk factors, such as altered serum lipid profile. Oxidative stress plays an important part in the pathogenesis of atherosclerosis. Serum antioxidants and intracellular enzymatic antioxidants composed mainly of glutathione peroxidase (GSH-Px), catalase (CAT), superoxide dismutase (SOD) and glutathione reductase counterbalance oxidative stress. Diminished activity of this system may lead to accelerated progression of atherosclerosis. The aim of this study was to assess the activity of CAT, GSH-Px, SOD and glutathione reductase in children with a family history of premature CHD who did not present any other major risk factors of CHD (diabetes, obesity, dyslipidaemia or hypertension). Twenty-two healthy children from high-risk families, selected according to the National Cholesterol Education Program definition, were enrolled in the study. The control group comprised 18 children without a family history of CHD. All the children were healthy and had been screened for hyperlipidaemia, diabetes, hypertension and obesity prior to the study. The erythrocyte activity of CAT, GSH-Px, SOD and glutathione reductase was assessed. Children at high risk of CHD had a statistically significant lower level of GSH-Px and CAT activity than the children in the control group. There were no statistically significant differences in the activity of SOD and glutathione reductase. PMID- 12751698 TI - A paracetamol-pasta test for assessing gastric emptying in healthy and diabetic subjects. AB - Previous studies have shown that the relationship between gastrointestinal symptoms and gastric emptying is weak. Therefore the quantitative assessment of gastric emptying with a relatively simple, non-invasive test would be of considerable clinical value in insulin-treated diabetic patients to identify those with disturbed gastric emptying. The aim of this investigation was to evaluate the inter- and intra-subject variability of a paracetamol-pasta test in healthy subjects and in IDDM patients. Eighteen healthy subjects (8 women) with a mean age of 37 years (range 19-68) and 19 IDDM patients (10 women) with a mean age of 48 years (range 25-62) and mean duration of diabetes of 28 years (range 6 52) were studied on two occasions with an interval of 1 to 4 weeks. After an overnight fast the subjects ingested a standardized pasta meal mixed with 2 g paracetamol in a period of 15 min. Blood samples were drawn at regular intervals after meal intake and analysed for paracetamol (P) and blood glucose. The serum levels of P were significantly lower at 15 min in diabetic patients. The intra subject coefficients of variation (CV%) of the areas under the serum paracetamol concentration-time curve (AUC) were almost identical in healthy and diabetic subjects, while the intra-subject CV of the P-Tmax was considerably lower in diabetic patients as well as markedly lower than the corresponding inter-subject CV. The inter-subject CVs of all parameters calculated were generally higher in diabetic patients. This study indicates that the assessment of paracetamol absorption kinetics during a paracetamol-pasta test is reproducible in healthy as well as in diabetic subjects. Diabetic patients with non-optimal glucose control and without a case history indicating gastroduodenal motor function disturbances achieve lower serum concentration of P at 15 min and generally display a higher inter-individual variability indicative of subclinical disturbances of gastric emptying in this group of patients. PMID- 12751699 TI - Inferior turbinectomy in children. AB - BACKGROUND: Inferior turbinectomy on patients of all ages is a controversial procedure. Its effect on children has been reported little in the literature and the few studies that are available involved relatively older children, i.e., >10 years old. Nasal obstruction caused by extensive hypertrophy of the inferior turbinates is not an uncommon observation in the pediatric population. The clinical manifestations might present as snoring, noisy breathing, mouth breathing, and, possibly, sleep apnea. METHODS: In this study, we followed 227 children <10 years of age who underwent inferior turbinectomy (27 children also underwent a revision of an earlier adenoidectomy), of whom 179 children had significant relief of nasal obstruction at the 1-year follow-up. RESULTS: Nocturnal breathing was reported to be more regular and otherwise improved in the 36 children with a suspected history of sleep apnea. Forty-two of 47 children who had thick nasal secretions and did not respond to antibiotic therapy before the operation had significant relief postoperatively. Postoperative complications were few and their number did not exceed that of adults. CONCLUSIONS: A complete inferior turbinectomy should be considered in children <10 years of age who have hypertrophied inferior turbinates that cause major interference with nasal breathing. PMID- 12751700 TI - Arterial ligation for pediatric epistaxis: developmental anatomy. AB - BACKGROUND: Anatomic studies of adult skulls have aided in the design of operations for the surgical ligation of nasal feeding vessels in the treatment of severe epistaxis. Lack of appropriate specimens has prevented similar studies in children. We performed an anthropometric study of archeological specimens to learn the effects of growth on key anatomic relationships. METHODS: We studied the skulls of children who died between 200 and 8000 years ago, recovered from archeological digs around the world. Measurements of the distances from the posterior lacrimal crest to the foramina of anterior and posterior ethmoidal arteries and optic canal and the pyriform aperture to the foramen of the sphenopalatine artery were made and compared with postnatal age, estimated from facial growth and dental eruption patterns. RESULTS: There is rapid growth in the orbit and midface during the first 6 years of life and gradual growth between 7 years and adulthood. The length of the medial wall of the orbit doubles during development with disproportionate enlargement of its anterior half. CONCLUSION: Arterial ligation is sometimes required for intractable pediatric epistaxis, especially after trauma. The changing relationships of critical structures in the orbital must be understood to allow safe ethmoidal artery ligation. The transantral approach to the maxillary artery is greatly limited by lack of midfacial development and maxillary pneumatization. We describe the necessary parameters for endoscopic, transnasal sphenopalatine artery ligation in growing children. PMID- 12751701 TI - Ventricular tachycardia as an electrocardiography artifact during functional endoscopic sinus surgery. AB - BACKGROUND: Functional endoscopic sinus surgery is frequently performed during conscious sedation not provided by an anesthesiologist. Several monitors are used during conscious sedation, one of which is the electrocardiogram (EKG). Equipment used in the operating room may interfere with the EKG tracing. METHODS: A 30-year old man underwent his fourth functional endoscopic sinus surgery during general anesthesia. For the procedure, a Xomed Straight Shot microdebrider was used. RESULTS: Because of stray electrical current, the use of the microdebrider caused the EKG waveform to resemble ventricular tachycardia. CONCLUSION: This case alerts the clinician to this possible occurrence to prevent inappropriate therapeutics from being initiated. PMID- 12751702 TI - Chronic sinonasal disease in patients with inflammatory bowel disease. AB - BACKGROUND: The objective of this study was to explore the possible relationship between inflammatory bowel disease (IBD) and chronic sinonasal disease. METHODS: A cross-sectional study was undertaken of 241 patients with Crohn's disease (CD) or ulcerative colitis from a tertiary medical center IBD clinic. Patient demographic data and information regarding IBD diagnosis and management, sinonasal disease diagnosis and management, and complications related to these diagnoses were gathered by retrospective chart review and a standardized patient survey. RESULTS: One hundred sixty surveys (67%) were returned and analyzed. Overall 48% of patients with IBD reported chronic sinonasal disease symptoms. Patients with CD had a higher prevalence of sinonasal disease than patients with ulcerative colitis (53% versus 32%; p < 0.02). The subgroup of CD patients with obstructive bowel complications had the highest prevalence of sinonasal disease (68% versus 27%; p = < 0.001), with 23% reporting chronic rhinosinusitis, 13% reporting chronic rhinitis, and an additional 32% reporting chronic nasal or sinus symptoms. CONCLUSION: The prevalence of chronic sinonasal disease is elevated in patients with IBD, occurring in approximately one-half of patients followed at a tertiary IBD center. Patients with CD experiencing obstructive complications had significantly increased rates of sinonasal disease. The relationship between chronic sinonasal disease and obstructive CD is not defined, but several hypotheses are generated. PMID- 12751703 TI - Spontaneous nasal cerebrospinal fluid leaks and empty sella syndrome: a clinical association. AB - BACKGROUND: Spontaneous, idiopathic nasal meningoencephaloceles are herniations of arachnoid/dura and cerebrospinal fluid (CSF) through anatomically fragile sites within the skull base. Empty sella syndrome occurs when intracranial contents herniate through the sellar diaphragm filling the sella turcica with CSF and giving the radiographic appearance of an absent pituitary gland. The objective of this study was to examine the association between spontaneous encephaloceles/CSF leaks and empty sella syndrome because of their similar clinical features and potential common pathophysiology. METHODS: Retrospective. RESULTS: Sixteen patients were treated for spontaneous encephaloceles between 1996 and 2001. All 16 patients had associated CSF leaks. Five patients had multiple simultaneous encephaloceles. Fifteen patients with imaging of the sella turcica had empty (10 patients) or partially empty (5 patients) sellas. One patient did not have complete imaging of the sella. Three patients had lumbar punctures with measurement of CSF pressure during computed tomography cisternograms because of multiple skull base defects. Mean CSF pressure was 28.3 cm of water (range, 19-34 cm; normal, 0-15 cm). Thirteen of 16 patients (81%) were obese women (mean body mass index 35.9 kg/m2; normal, <25 kg/m2). Mean follow-up was 14.2 months with 100% success in closure of the defects after one procedure. CONCLUSION: Spontaneous meningoencephaloceles and CSF leaks are strongly associated with radiographic findings of an empty sella and suggest a common pathophysiology. The underlying condition probably represents a form of intracranial hypertension that exerts hydrostatic pressure at anatomically weakened sites within the skull base. Otolaryngologists should be familiar with this disease entity and the implications intracranial hypertension has on patient management. PMID- 12751704 TI - Management of the orbital floor in silent sinus syndrome. AB - BACKGROUND: Enophthalmos in a patient with an opacified hypoplastic maxillary sinus, without sinus symptomatology, describes the silent sinus syndrome. A current trend is to perform endoscopic maxillary antrostomy and orbital floor reconstruction as a single-staged operation. A two-staged approach is performed at our institution to avoid placement of an orbital floor implant in the midst of potential infection and allow for the possibility that enophthalmos and global ptosis may resolve with endoscopic antrostomy alone, obviating the need for orbital floor reconstruction. METHODS: A retrospective review identified four patients with silent sinus syndrome evaluated between June 1999 and August 2001. Patients presented to our ophthalmology department with ocular asymmetry, and computerized tomography (CT) scanning confirmed the diagnosis in each case. RESULTS: There were three men and one woman, with ages ranging from 27 to 40 years. All patients underwent endoscopic maxillary antrostomy. Preoperative enophthalmos determined by Hertel's measurements ranged from 3 to 4 mm. After endoscopic maxillary antrostomy, the range of reduction in enophthalmos was 1-2 mm. Case 2 had a preoperative CT scan and a CT scan 9 months after left endoscopic maxillary antrostomy. Volumetric analysis of the left maxillary sinus revealed a preoperative volume of 16.85 +/- 0.06 cm3 and a postoperative volume of 19.56 +/- 0.07 cm3. This represented a 16% increase in maxillary sinus volume postoperatively. Orbital floor augmentation was avoided in two patients because of satisfactory improvement in enophthalmos. In the other two patients, orbital reconstruction was performed as a second-stage procedure. There were no complications. CONCLUSION: Orbital floor augmentation can be offered as a second stage procedure for patients with silent sinus syndrome. Some patients' enophthalmos may improve with endoscopic antrostomy alone. PMID- 12751705 TI - Phase contrast microscopy with modified Hansel's staining: mast cell identification and activity-related changes in cell membrane refractivity in allergic nasal smears. AB - BACKGROUND: A new technique called phase contrast microscopy with modified Hansel's staining (used with bright field microscopy) was developed to identify mast cells (MCs) and granulocytes (eosinophil/neutrophil/basophil [E/N/B]). METHODS: Nasal scratching smears from 618 patients with Japanese cedar pollinosis were examined using this new technique. RESULTS: This technique permitted accurate morphological identification. MCs can be discriminated from E/N/Bs. The surface of the cell membrane appeared as low refractile (lr), moderately refractile (mr), or high refractile (hr). This was caused by the light, which is not related to the phase difference, but rather originated from the difference in the refraction of the direct light. In specimens from the onset stage (i.e., 1-3 days after onset), lr-MC and mr/hr-E/N/B were dominant. In specimens from the early stage (i.e., 4-7 days after onset), lr-E/N/B significantly increased in number. CONCLUSION: The phospholipid bilayers of the cell membrane exhibit a phase transition after the onset, and phase refractivity of the cell membrane is closely related to the activity of the cell. This indicates that in the onset stage, MCs are already activated, whereas most of the E/N/Bs are not. In contrast, the latter cell types become activated subsequently in the early stage. PMID- 12751706 TI - The four components of the nasal valve. AB - The nasal valve consists of four distinct airflow-resistive components. (i) The vestibule terminates in an airflow-resistive aperture between the septum and the caudal end of the upper lateral cartilage. Its cross-sectional area is stabilized by the cartilaginous structures and by inspiratory isometric contractions of alar dilator muscles. Its walls are devoid of erectile tissues that might otherwise affect its cross-sectional area and airflow resistance. By contrast, (ii) the bony entrance to the cavum is occupied by erectile tissues of both (iii) lateral (turbinate) and (iv) septal nasal walls that modulate the cross-sectional area of the airway and airflow resistance. The body of the cavum offers little resistance to airflow. Valve constrictions induce "orifice flow" of inspiratory air as it enters the body of the cavum, disrupting laminar characteristics and thereby enhancing exchanges with the nasal mucosa of heat, water, and contaminants. Acoustic rhinometric and rhinomanometric measurements show the sites, dimensions, and resistances of the valve constrictions and indicate that it is seldom necessary to extend septal and/or turbinate surgery far beyond the piriform aperture in the treatment of nasal obstruction. PMID- 12751707 TI - An association between acquired epiphora and the signs and symptoms of chronic rhinosinusitis: a prospective case-control study. AB - BACKGROUND: It has been suggested that chronic rhinosinusitis may lead to epiphora because of inflammatory edema at the nasal end of the nasolacrimal duct and that treatment of the underlying nasal disease may obviate the need for dacryocystorhinostomy. The aim of this study was to establish whether or not there is an association between the signs and symptoms of chronic rhinosinusitis and a complaint of epiphora in a blinded, prospective case-control study. METHODS: A consecutive series of 15 adult patients presenting to the ophthalmology department with acquired epiphora were compared with 29 patients presenting with chronic open-angle glaucoma over the same study period. Nasal symptoms and the findings on nasal endoscopy were recorded by a single otolaryngologist blinded to the diagnosis and using a standardized staging system RESULTS: The age and sex distributions of the two groups were similar. Scores for headache and altered smell were significantly higher in the epiphora cases than in controls (p = 0.05 and 0.03, respectively). Trends for higher scores for congestion and discharge were not statistically significant (p = 0.06 and 0.07, respectively). Mucosal edema and discharge were significantly more common on endoscopy in the epiphora cases than in the control cases (p < 0.02). CONCLUSION: We have shown an association between rhinosinusitis and acquired epiphora, and this would be consistent with chronic rhinosinusitis being the cause of the epiphora in some cases. PMID- 12751708 TI - Image-guided resection of fibro-osseous lesions of the skull base. AB - BACKGROUND: Endoscopic resection of sinonasal fibro-osseous lesions remains a technical challenge because of the loss of anatomic landmarks and the frequent need to resect bone along the skull base. The purpose of this study was to evaluate the usefulness of image-guidance systems for the resection of these lesions. METHODS: Endoscopic surgery was performed in 10 patients with fibro osseous lesions of the sinuses and skull base (six fibrous dysplasias and four osteomas). Indications for surgical intervention were recurrent infection (7), facial pain (3), mucocele (1), and compressive optic neuropathy (1). All surgeries were performed with an image-guidance system, which included an integrated high-speed drill system. RESULTS: Anatomic localization was accurate to within 1 mm when monitoring the depth of bone removal along the skull base. Complete resection of the fibro-osseous lesion was possible in six patients (60%) and partial removal was possible in four patients (40%). In the latter group, a margin of dysplastic bone was preserved along the ethmoid roof to prevent dural exposure. There were no intraoperative complications. Surgery resulted in resolution of infection and facial pain in nine patients (90%). Additional surgery was required in one patient (10%) who developed recurrent optic neuropathy because of continued dysplastic bone growth. Mean follow-up was 34 months (range, 13-67 months). CONCLUSION: Image-guidance technology appears to be ideally suited for the treatment of patients with fibro-osseous lesions of the sinonasal cavity. Real-time monitoring of the depth of bone removal relative to the skull base may enhance the safety and efficacy of such surgery. PMID- 12751709 TI - Visual loss following injection. PMID- 12751710 TI - Antiretroviral treatment of human immunodeficiency virus infection: Swedish recommendations. AB - The Swedish guidelines (SwG) for treatment of human immunodeficiency virus (HIV) infection have several important roles. A major task involves the promotion of a uniformly high standard of care in all HIV treatment clinics in Sweden and the identification of strengths, weaknesses and relevance of recent research findings. CD4+ T-cell counts < 200 cells/microl are clear indications for the initiation of treatment, whereas high viral loads serve as an indication for increased vigilance rather than a criterion for therapy. It is recommended that the first regimen consists of 2 nucleoside reverse transcriptase inhibitors in combination with 1 protease inhibitor or 1 non-nucleoside reverse transcriptase inhibitor. The definition of treatment failure is rigorous. Treatment change should be considered if the viral load has not fallen by at least 1.5 log in 4 weeks or is undetectable within 3-4 months. Resistance testing is endorsed at primary infection, in the event of treatment failure and in pregnant women. Interaction with experts in HIV resistance testing is emphasized. Therapeutic drug monitoring is advocated. Patients with treatment failure should be handled individually and the decision on therapeutic strategy should be based on treatment history, resistance testing and other clinical facts. The SwG do not give recommendations for some important issues such as prolonged drug holidays and preferences in initial treatment regimens. More scientific data are likely to be available soon and the SwG will be refined accordingly. The present guidelines are translated from Swedish; they are published on the Medical Products Agency (MPA) and Swedish Reference Group for Antiviral Therapy (RAV) websites (www.mpa.se and www.rav.nu.se), including 7 separate papers based on a thorough literature search. A complete reference list is available on request from the MPA. PMID- 12751711 TI - A novel thiocationic liposomal formulation of antisense oligonucleotides with activity against Mycobacterium tuberculosis. AB - This study describes the development of a novel thiocationic (OBEHYTOP) lipid based formulation of phosphorothioate antisense oligonucleotides (PAOs) showing inhibitory activity against mycobacterium tuberculosis (mTB) as measured by an in vitro BACTEC 460TB assay. PAOs were designed based on sequences complementary to essential regions of the mycobacterial genome from published nucleic acid databases in GenBank. These included the superoxide dismutase sod A gene (TBS3), catalase-peroxidase katG gene (TBK1, TBK10), RNA polymerase beta-subunit rpo B gene (TBR5) and diaminopimelate decarboxylase lys A gene (TBL5). The effect of PAOs (TBS3, K1, K10, R5 and L5) alone on mTB was not significant compared with the no-drug control over a period of exposure of 150 h (ranges of -11.8 to +23.58% at 72 h; 15.26 to +25.82% at 96 h and -5.51 to +24.00% at 150 h). Liposomal formulations (10:5:2 OBEHYTOP:oleic acid:vitamin D3) of PAOs resulted in statistically significant (p < 0.05 in all cases) inhibition (ranges of -51.45 to -63.00% at 72 h; -56.75 to -67.96% at 96 h; -51.45 to -60.26% at 150 h) compared with PAOs alone, thiocationic liposomal control and liposomal components. Positive controls of streptomycin and isoniazid used at their minimum inhibitory concentrations of 2.00 and 0.10 microM, respectively, resulted in average % inhibition values of -94% and -97.36%, respectively, indicating that these thiocationic lipid-formulated PAOs showed inhibitory activity directed against mTB in vitro. PMID- 12751712 TI - Safety and efficacy of molgramostim as an adjunctive therapy in critically ill patients with severe sepsis. AB - The aim of this uncontrolled, prospective, clinical study was to investigate the efficacy and safety of molgramostim administration in patients with severe sepsis. The subjects were 20 critically ill, mechanically ventilated patients with severe sepsis in a university intensive care unit (ICU). Molgramostim 300 microg s.c. was given every 12 h for 3 d. Treatment for severe sepsis was also administered as medically indicated. No adverse events (clinical or serum chemistry) were considered as drug related. Temperature (p = 0.334) and PaO2/FiO2 index (arterial oxygen tension/inspiratory oxygen fraction) (p = 0.178) were not significantly changed. Total leukocyte and neutrophil count increased significantly (p < 0.001) during drug administration. Simplified Acute Physiology Score II (SAPS II) was not significantly increased (p = 0.955), but there was a statistically significant decrease (p = 0.006) in Sepsis-related Organ Failure Assessment (SOFA) score. Death probability was not statistically different compared with mortality rate on day 28 and overall mortality (p = 0.238 and 0.700, respectively). There were statistically significant decreases (p < 0.01) in serum tumor necrosis factor-alpha (TNF-alpha), TNF-RII and interleukin-2 (IL 2), and an increase in TNF-RI levels between study entry and day 3. Mean ICU stay was 40.2 +/- 7.7 d. In conclusion, molgramostim administration may not affect serum chemistry and PaO2/FiO2 index, may decrease SOFA score but does not produce significant clinical benefit in terms of patients' outcome compared with death probability. It may also influence TNF-alpha, TNF-RI and TNF-RII serum complex levels. These changes may be attributed to the natural clinical course of sepsis or therapy applied. PMID- 12751713 TI - Hepatitis C virus RNA levels during initial induction or standard interferon therapy: influence of continued treatment on sustained response. AB - Hepatitis C virus (HCV) RNA kinetics were studied at baseline weeks 4, 8 and 12 during interferon-alpha (IFN) monotherapy in 65 patients (mean age 39 y, range 19 66 y) with chronic HCV infection. IFN treatment was given either as initial induction (n = 34) or as standard dosing 3 times a week (n = 31). Patients with genotypes 2 and 3 had a significantly steeper decline in HCV RNA levels than patients with genotype 1 at weeks 4, 8 and 12 (p < 0.001 at all points measured). The decline in viral load was more pronounced in patients with induction therapy than with standard therapy at weeks 4, 8 and 12 (p < 0.02, 0.054 and 0.01, respectively). Patients with a sustained viral response had a 3-log decline in viral levels at week 4, with few exceptions. Two patients with non-response at week 12 (1 each with genotype 1 and non-1) responded after supplementation with ribavirin. PMID- 12751714 TI - Symptomatic intestinal amoebiasis and climatic parameters. AB - Symptomatic intestinal amoebiasis is an important problem, especially in the developing world. The relationships between climatic parameters and amoebic dysentery cases were evaluated in this study. Climatic data were obtained from the local meteorological department and the diagnosis of amoebic dysentery was established by clinical and laboratory investigation. Monthly mean temperature (r = 0.755, p = 0.005), monthly mean maximum temperature (r = 0.711, p = 0.01), monthly mean temperature at 100 cm underground (r = 0.818, p = 0.001) and monthly mean humidity values (r = -0.656, p = 0.02) correlated significantly with symptomatic disease. Although inverse relationships were found for humidity and atmospheric pressure, the monthly mean atmospheric pressure (r = -0.084) did not seem to have a significant effect on intestinal amoebiasis (p = 0.80). Thus, when the weather warmed up, the frequency of symptomatic intestinal amoebiasis increased significantly. To improve the ability to predict disease trends, it seems logical to assess the independent and interactive effects of climatic parameters on disease impact. PMID- 12751715 TI - Disseminated Nocardia transvalensis infection with osteomyelitis and multiple brain abscesses. AB - A case of Nocardia transvalensis osteomyelitis and cerebral abscesses is described. To the authors' knowledge, the patient described here represents the first reported case of primary N. transvalensis leg abscess with contiguous osteomyelitis of the femur and secondary brain abscesses. The literature on nocardial osteomyelitis and cerebral nocardiosis is reviewed, and treatment of both conditions is discussed. The interactions between the immune system and nocardial organisms are described, as well as the host defense defects in diabetic patients as they relate to pathogenicity of the bacteria. Finally, the patient's paradoxical therapeutic response of resolving brain abscesses with concomitant unresolving osteomyelitis is discussed, as well as its implications for managing similar cases in the future. PMID- 12751716 TI - Spontaneous bacterial peritonitis due to Brucella melitensis. AB - Peritonitis is an extremely rare complication of brucellosis. A case is reported of blood and ascitic culture-proven spontaneous bacterial peritonitis caused by Brucella melitensis, in a patient who had also cirrhosis. PMID- 12751717 TI - Swyer-James (MacLeod's) syndrome following pertussis infection in an infant. AB - Pertussis is a potentially severe disease, especially in infancy. The case of an 8-month-old infant is described who presented the typical radiographic image of unilateral hyperlucent lung or Swyer-James (MacLeod's) syndrome. The infant suffered from pertussis at 20 d of age. A rare postinfectious complication of pertussis is reported. PMID- 12751718 TI - Repeated gallium-67 scan demonstrating an occult mycotic aneurysm of the aortic arch due to Salmonella. AB - A 71 y-old female with 40 degrees C fever and a change in consciousness during haemodialysis was referred for a gallium-67 scan, which revealed a horseshoe-like radioactive accumulation in the mediastinal region. Salmonella enteritidis was isolated from blood culture. Magnetic resonance imaging confirmed an aortic arch aneurysm. The patient died without surgery. PMID- 12751719 TI - Citrobacter koseri (diversus) meningitis in an otherwise healthy adolescent. AB - Citrobacter infection is commonly reported in neonates and immunocompromised patients. Citrobacter koseri (diversus) is an important cause of neonatal meningitis and brain abscess formation. It adults, however, Citrobacter infection with central nervous system involvement is rare, and is usually associated with an underlying disorder. This report describes a 12-y-old previously healthy girl with Citrobacter koseri meningitis. Intravenous treatment with ceftriaxone for 10 d led to complete recovery. Head computed tomography and brainstem-evoked response audiometry were normal. On follow-up, the patient was completely healthy. Previously reported cases of C. koseri meningitis in the adult population were associated with underlying predisposing factors. In this case a normal, healthy adolescent was treated with intravenous ceftriaxone without any of the known neurological complications observed in the neonatal cases. PMID- 12751720 TI - Acute abdomen due to Brucella melitensis. AB - A case of acute abdomen caused by a Brucella melitensis is reported. The patient presented with biliary involvement in the form of acute acalculous cholecystitis and developed acute appendicitis that resulted in his surgical treatment. PMID- 12751721 TI - Streptococcus zooepidemicus meningitis and bacteraemia. AB - Group C streptococci are common causative agents of epidemic infections in animals and a rare cause of meningitis in humans. The case is reported of a 75-y old man with meningitis caused by a group C streptococcus (Streptococcus zooepidemicus). He had frequent contact with horses, which were a possible source of infection. In spite of treatment with a third generation cephalosporin, the outcome was fatal. PMID- 12751722 TI - Acanthamoeba keratitis in a non-contact lens wearer with human immunodeficiency virus. AB - Acanthamoeba keratitis is potentially blinding and often associated with contact lens wearing. A human immunodeficiency virus (HIV)-positive patient, a non contact lens wearer, presented with keratitis. She experienced a protracted course of disease, characterized by exacerbations and remissions, and was treated with various topical antibiotics and steroids. 13 months after symptom onset the eye was removed owing to serious scarring of cornea and unbearable pain. Microbiological and histopathological examination of the cornea showed Acanthamoeba. In non-contact lens wearers suffering from Acanthamoeba keratitis the diagnosis is delayed, pathognomonic features are often not seen and visual outcome is usually poor. There is no known relation between HIV infection and Acanthamoeba keratitis. PMID- 12751723 TI - Transmission of human immunodeficiency virus and hepatitis B virus by blood brotherhood rituals. AB - Blood brother/sisterhood is a ritual that involves the exchange of a small amount of venous blood via the binding of excisions. Human immunodeficiency virus (HIV) and hepatitis B virus (HBV) could be transmitted by this ritual. Two cases infected with HIV and HBV by a blood brotherhood ritual are reported. PMID- 12751724 TI - Chronic diarrhoea caused by Cryptococcus neoformans in a non-human immunodeficiency virus-infected patient. AB - A 43-y-old woman with underlying cervix carcinoma stage IIIB, status postirradiation, and diabetes mellitus with tripathy suffered from chronic diarrhoea for more than 6 months. Stool examination showed few white blood cells and red blood cells. Rectal swab cultures disclosed Cryptococcus neoformans in 2 samples obtained 2 weeks apart. The diarrhoea responded to treatment with oral fluconazole for 4 weeks. Repeated rectal swabs revealed no yeast growth. During follow-up for 4 y, she had no problems with diarrhoea. Chronic diarrhoea caused by C. neoformans is rare and clinical suspicion is needed. PMID- 12751725 TI - A case of tick-transmitted lymphadenopathy in Bulgaria associated with Rickettsia slovaca. PMID- 12751727 TI - Leukotriene receptors: state of the art. PMID- 12751726 TI - Leukotrienes in asthma. PMID- 12751728 TI - LTC4 production by eosinophils in asthmatic subjects with alternative forms of ALOX-5 core promoter. PMID- 12751729 TI - Il-4-induced lipid mediators class switching in human normal mononuclear phagocytes. PMID- 12751730 TI - Lipoxins and aspirin-triggered lipoxins in airway responses. AB - Here, we have demonstrated potent inhibition of allergen-mediated pulmonary inflammation and airway hyper-responsiveness by a novel dual-pronged action of LX's. Moreover, these results indicate that LX's play pivotal and previously unappreciated roles in regulating allergy and pulmonary inflammation. PMID- 12751731 TI - Potential role of cysteinyl leukotrienes in trafficking and survival of hematopoietic progenitor cells. PMID- 12751732 TI - Covalent binding of leukotriene A4 to nucleosides, nucleotides, and nucleic acids. PMID- 12751733 TI - Firm adhesion of neutrophils to cerebral vascular endothelium "in vivo": a role for cys-leukotrienes. PMID- 12751734 TI - The CYP P450 arachidonate monooxygenases: enzymatic relays for the control of kidney function and blood pressure. PMID- 12751735 TI - Cytochrome P450-derived eicosanoids mediators of ocular surface inflammation. are. PMID- 12751736 TI - Regulation of renal microvascular 20-hydroxyeicosatetraenoic acid (20-HETE) levels. PMID- 12751737 TI - Cyclooxygenase activity is regulated by the heme oxygenase system in microvessel endothelial cells. PMID- 12751739 TI - Thromboxane-induced ERK phosphorylation in human aortic smooth muscle cells. PMID- 12751738 TI - Cytochrome P450 arachidonic acid metabolites modulate renal tubular function. PMID- 12751740 TI - Arachidonic acid inhibits cysteinyl-leukotriene receptor activation in human pulmonary vessels. PMID- 12751741 TI - Cytosolic phospholipase A2alpha in mouse platelets. PMID- 12751742 TI - Cellular arachidonate-releasing functions of various phospholipase A2s. PMID- 12751743 TI - Biological functions of group X secretory PLA2. PMID- 12751744 TI - Protection of cell membrane from exogenous PLA2 and related inflammatory stimuli by membrane-anchored lipid conjugates. PMID- 12751745 TI - Regulatory functions of prostaglandin E2 synthases. PMID- 12751746 TI - A novel type of membrane-associated prostaglandin E synthase. PMID- 12751747 TI - Expression, purification and characterization of recombinant human microsomal PGE2 synthase-1. PMID- 12751748 TI - Formation of prostaglandin E2, leukotriene B4 and 8-isoprostane in alveolar macrophages by ultrafine particles of elemental carbon. PMID- 12751749 TI - Effects of PGF2alpha analogues in experimental morphine-induced pharmacodependence. PMID- 12751750 TI - Inhibition of prostaglandin H2 synthases by salicylate is dependent on the oxidative state of the enzymes. PMID- 12751751 TI - 5-Lipoxygenase activation by MAPKAPK-2 and ERKs. PMID- 12751752 TI - Renal effects of selective cyclooxygenase inhibition in experimental liver disease. PMID- 12751753 TI - Variable antipyretic effect of SC-58236, a selective cyclooxygenase (COX)-2 inhibitor, in rats. PMID- 12751754 TI - Induction of cyclooxygenase-2 expression by fluid smear stress in vascular endothelial cells. PMID- 12751755 TI - Induction of COX-2 expression by the endocannabinoid derivative R(+) methanandamide. PMID- 12751756 TI - Specific gene expression of isoforms in cyclooxygenase pathway to generate PGJ2 derivatives during the differentiation of 3T3-L1 adipocytes. PMID- 12751757 TI - Tristetraprolin binds to the COX-2 mRNA 3' untranslated region in cancer cells. PMID- 12751758 TI - Platelet activating factor receptors. PMID- 12751759 TI - Splicing mutation of the prostacyclin synthase gene in a family associated with hypertension. PMID- 12751760 TI - DNA methylation regulates 5-lipoxygenase promoter activity. PMID- 12751761 TI - Isomerization and nitro-oxidation of arachidonic acid by NO2. PMID- 12751762 TI - Prostaglandin F2alpha protected cultured Madin-Darby canine kidney cells from the development of apoptosis induced by 12-o-tetradecanoyl phorbol-beta-acetate and stimulated synergistically with nordihydroguaiaretic acid. PMID- 12751763 TI - LDL receptor-related protein plays an essential role in 12/15-lipoxygenase mediated LDL oxidation by macrophages. PMID- 12751764 TI - Effects of in vitro exposure to carbon dioxide on the expression of apoptosis associated genes in human peritoneal fibroblasts. PMID- 12751765 TI - Identification, characterization, and properties of a class alpha microsomal glutathione S-transferase. PMID- 12751766 TI - Cyclooxygenase (COX) function in the ductus arteriosus: another look. PMID- 12751767 TI - Evidence for enhanced oxidative stress in coronary heart disease and chronic heart failure. PMID- 12751768 TI - The leukotriene receptor CysLT1 and 5-lipoxygenase are upregulated in colon cancer. PMID- 12751769 TI - Increased expression of 5-lipoxygenase in glioblastoma multiforme. PMID- 12751770 TI - Variable influence of statins on isoprostanes in hyperlipidemia. PMID- 12751771 TI - Increased isoprostanes in children of smoking parents. PMID- 12751772 TI - The dinor isoprostane pathway in plants. PMID- 12751774 TI - Cryopreservation of oocytes from pre-antral follicles. AB - Cryobiology is a very important tool in reproductive biology. Research in this area focuses on the possibility of restoring fertility in women with reproductive problems or after cancer treatments. Another goal is to establish a genetic resource bank for endangered or commercially important animal species. Cryopreservation of oocytes from pre-antral follicles has been studied during the past decade. Procedures can be divided between the cryopreservation of either ovarian tissue or isolated follicles. Most studies describe a slow freezing/rapid thawing protocol to cryopreserve ovarian fragments. Histology shows that the follicles maintain their morphological integrity, and transplantation of ovarian tissue demonstrates that the follicles can restart their growth and eventually ovulate. Some research groups have obtained offspring using this procedure in mice and sheep. With regard to the cryopreservation of isolated follicles, the few studies reported in this area used the same freezing protocol, and some of them described follicular growth using in-vitro culture. The best result was obtained in mice, with animal birth after follicular cryopreservation and culture. However, additional studies are necessary for a better understanding of the events during follicular cryopreservation and to establish a standard protocol for ovarian transplantation or follicle culture. PMID- 12751773 TI - Progesterone as a mediator of gonadotrophin action in the corpus luteum: beyond steroidogenesis. AB - Studies using newer, potent GnRH antagonists and pure gonadotrophins have clarified the importance of: (i) the strength-duration of the midcycle surge of pituitary gonadotrophins (LH, FSH) in follicle rupture and conversion to the corpus luteum; (ii) the continued requirement for pituitary LH throughout development and the functional lifespan of the primate corpus luteum in the menstrual cycle; and (iii) the exponential secretion of chorionic gonadotrophin (CG) by the developing placenta to extend the functional lifespan of the primate corpus luteum in early pregnancy. Although studies continue to increase current understanding of the cellular and molecular actions of LH/CG to stimulate luteal steroidogenesis, knowledge of the processes whereby these gonadotrophins promote the development and maintenance of the functional corpus luteum remains limited. This review summarizes evidence that the primate ovulatory follicle and corpus luteum is a target for the primary steroid produced by luteinizing/luteal tissue (i.e. progesterone). With evidence for dynamic expression of genomic progesterone receptors (PRA/B), and possibly other progesterone-receptor systems, recent studies addressed the hypothesis that progesterone is a critical 'local luteotrophin' that promotes luteal development and sustains luteal structure function during the menstrual cycle and early pregnancy. Specific progesterone actions to regulate tissue remodelling (via protease expression), health (anti apoptotic effects) and sensitivity to other local factors (e.g. via estrogen receptor expression) are discussed. The collective data suggest that there are gonadotrophin-stimulated, progesterone-dependent processes that promote luteotrophic and suppress luteolytic pathways in the primate corpus luteum. However, further studies are needed to verify their role in normal ovarian function and relevance to possible ovarian defects in natural and assisted reproduction technique-related cycles. PMID- 12751775 TI - Endometrial wave-like activity in the non-pregnant uterus. AB - The non-pregnant uterus shows wave-like activity throughout the menstrual cycle. This uterine activity was first detected using intra-uterine pressure recordings. The use of ultrasound has made it possible to study the movements of the uterus in a non-invasive manner. Throughout the menstrual cycle, several wavelike activity patterns have been described; these patterns change throughout the menstrual cycle and are governed by steroid hormones. An adequate wave pattern seems to be related to successful reproduction in spontaneous cycles and assisted reproduction. Further insight into the phenomenon of endometrial wave-like activity might offer an opportunity to correct abnormal wave patterns and thereby improve pregnancy rates. PMID- 12751776 TI - Uterine adenomyosis in the infertility clinic. AB - This review discusses uterine adenomyosis as a (co-)factor in female subfertility. The clinical presentation of adenomyosis uteri is reviewed as well as recent developments in non-invasive imaging modalities for the condition. Different treatment options are discussed, focusing on conservative management in patients who wish to maintain their childbearing capacity. PMID- 12751777 TI - Thyroid autoimmunity and hypothyroidism before and during pregnancy. AB - In the present review, an attempt was made to describe current knowledge and concepts concerning the complex relationships that link thyroid autoimmunity (TAI) and hypothyroidism with female and male infertility, as well as abnormalities occurring during pregnancy, such as pregnancy loss and maternal and fetal repercussions associated with hypothyroidism. In the case of infertility, although the clinical relevance of TAI is somewhat controversial, when all available information is considered the results strongly suggest that when infertility is due to well-defined female causes, autoimmunity is involved and TAI constitutes a useful marker of the underlying immune abnormality, independently of thyroid function disorders. In the case of pregnancy loss, the vast majority of available studies clearly establish that TAI (even with no overt thyroid dysfunction) is associated with a significant increase in miscarriage risk. To find an association, however, does not imply a causal relationship, and the aetiology of increased pregnancy loss associated with TAI remains presently not completely understood. With regard to maternal repercussions during gestation, the main risk associated with TAI is the occurrence of hypothyroidism and obstetric complications (premature birth, pre-eclampsia, etc.). Thus, systematic screening of TAI and hypothyroidism during early pregnancy, monitoring of thyroid function with/without L-thyroxine treatment and follow-up during post partum have proved helpful and important in order to manage these patients adequately. Finally, with regard to potential repercussions affecting the offspring, recent evidence suggests that thyroid maternal underfunction, even when considered mild (or subclinical), may be associated with an impairment of fetal brain development. When present only during the first half of gestation, maternal hypothyroxinaemia is a risk factor for impaired fetal brain development, due to insufficient transfer of maternal thyroid hormones to the feto-placental unit. When hypothyroidism is not restricted to the first trimester and worsens as gestation progresses (as in untreated hypothyroidism), the fetus may also be deprived of adequate amounts of thyroid hormones during later neurological maturation and development, leading to poorer school performance and lower IQ. PMID- 12751778 TI - A review of immune cells and molecules in women with recurrent miscarriage. AB - Immunological rejection of the fetus due to recognition of paternal antigens by the maternal immune system, resulting in abnormal immune cells and cytokine production, is postulated to be one cause of unexplained pregnancy loss. Although there is evidence for this in rodents, there is less evidence in humans. This article focuses on studies in humans, and reviews the recent literature on the differences in immune cells and molecules in normal fertile women and women with recurrent miscarriage (RM). Although much of the evidence is contradictory, these studies do suggest differences in the expression of some immune cells and molecules in women with RM. Differences in the CD56+ population of cells are seen, and there is some evidence for an alteration in the ratio of Th1 and Th2 cytokines produced by peripheral blood monocytes (PBMCs) and clones of decidual CD4+ cells. There is also some evidence for differences in endometrial cytokine production, and in particular decreased production of pro-inflammatory cytokines such as interleukin-6. Possible reasons for the variations in data are discussed, and the importance of compartment (peripheral blood, endometrium or decidua) in which the cells and molecules are measured and the timing of the sampling, both with respect to the menstrual cycle and pregnancy (at the time or just after miscarriage) is emphasized. PMID- 12751779 TI - Pathophysiology of increased nuchal translucency: a review of the literature. AB - First-trimester nuchal translucency (NT) measurement is a widely used technique to determine the risk of fetal aneuploidy. The aim of this review was to explore the different hypotheses on the aetiology of increased NT. Using a computerized database (PubMed), articles on the aetiology of first-trimester NT were retrieved. Furthermore, the cited references of the studied articles were used to find additional articles. Based on ultrasonographic and post-mortem morphological studies, the findings in increased NT fetuses are described and classified into three categories. First, an association between increased NT and cardiac abnormalities, combined with abnormal ductus venosus flow velocities, is found, leading to the theory that cardiac failure causes NT enlargement. Second, various types of abnormalities are found in the extracellular matrix of the nuchal skin of fetuses with increased NT. Third, abnormal lymphatic development is demonstrated in fetuses with increased NT. In conclusion, many hypotheses on NT enlargement are based on associations and speculations. This report provides an overview of the different theories and an attempt to fit the different associations and findings into a single hypothesis of abnormal embryological development. PMID- 12751780 TI - Results of conservative management of epithelial malignant and borderline ovarian tumours. AB - Conservative management of at least part of both the ovary and uterus can be proposed in patients with borderline ovarian tumour, in order to preserve fertility potential. This conservative management could be carried out even in patients with borderline ovarian tumour associated with non-invasive peritoneal implants (if complete resection of peritoneal disease has been performed). When facing persistent infertility after this conservative surgery, ovarian induction or an in-vitro procedure could be proposed in patients with an early-stage disease, though the number of attempts must be limited. Removal of the preserved ovary after completion of pregnancy(ies) is unnecessary if patients agree to careful follow-up. In patients with epithelial ovarian cancer, conservative management could safely be performed in young patients who wish to preserve fertility function and who fulfil the following criteria: unilateral tumour (stage IA), grade 1 (and 2?), adequate staging surgery and careful follow-up. Removal of the preserved ovary should be carried out after completion of pregnancy(ies) in order to reduce the risk of ovarian tumour recurrence. PMID- 12751781 TI - Glutathione pathways in the brain. AB - The antioxidant glutathione (GSH) is essential for the cellular detoxification of reactive oxygen species in brain cells. A compromised GSH system in the brain has been connected with the oxidative stress occuring in neurological diseases. Recent data demonstrate that besides intracellular functions GSH has also important extracellular functions in brain. In this respect astrocytes appear to play a key role in the GSH metabolism of the brain, since astroglial GSH export is essential for providing GSH precursors to neurons. Of the different brain cell types studied in vitro only astrocytes release substantial amounts of GSH. In addition, during oxidative stress astrocytes efficiently export glutathione disulfide (GSSG). The multidrug resistance protein 1 participates in both the export of GSH and GSSG from astrocytes. This review focuses on recent results on the export of GSH and GSSG from brain cells as well as on the functions of extracellular GSH in the brain. In addition, implications of disturbed GSH pathways in brain for neurodegenerative diseases will be discussed. PMID- 12751782 TI - The role of glutathione peroxidases in trypanosomatids. AB - Reactive oxygen species are the unwanted by-products of aerobic metabolism. To protect cells against their potentially lethal effects a series of pathways have evolved that are collectively called the oxidative defence system. In most eukaryotes, catalases and selenium-dependent glutathione peroxidases form the front line of defence against hydroperoxide-mediated damage. However, these activities are lacking in members of the Trypanosomatidae family of protozoan parasites. Instead these organisms contain several enzyme-mediated pathways for removal of hydroperoxides that are centred upon the unusual thiol trypanothione. Here we discuss the biochemical properties of one group of these enzymes, the non selenium glutathione-dependent peroxidases, and outline the roles that they play in protecting the parasite against hydroperoxides associated with biological membranes. PMID- 12751783 TI - Cytoprotection against oxidative stress and the regulation of glutathione synthesis. AB - Adaptation to oxidative and nitrosative stress occurs in cells first exposed to a nontoxic stress, resulting in the ability to tolerate a toxic challenge of the same or a related oxidant. Adaptation is observed in a wide variety of cells including endothelial cells on exposure to nitric oxide or oxidized lipids, and lung epithelial cells exposed to air-borne pollutants and toxicants. This acquired characteristic has been related to the regulation of a family of stress responding proteins including those that control the synthesis of the intracellular antioxidant glutathione. The focus of this article, which includes a review of recent results along with new data, is the regulation and signaling of glutathione biosynthesis, especially those relating to adaptive mechanisms. These concepts are illustrated with examples using nitric oxide and oxidized low density lipoprotein mediated adaptation to oxidative stress. These data are discussed in the context of other adaptive mechanisms relating to glutathione synthesis including those from dietary constituents such as curcumin. PMID- 12751784 TI - The parasite-specific trypanothione metabolism of trypanosoma and leishmania. AB - The bis(glutathionyl)spermidine trypanothione exclusively occurs in parasitic protozoa of the order Kinetoplastida, such as trypanosomes and leishmania, some of which are the causative agents of several tropical diseases. The dithiol is kept reduced by the flavoenzyme trypanothione reductase and the trypanothione system replaces in these parasites the nearly ubiquitous glutathione/glutathione reductase couple. Trypanothione is a reductant of thioredoxin and tryparedoxin, small dithiol proteins, which in turn deliver reducing equivalents for the synthesis of deoxyribonucleotides as well as for the detoxification of hydroperoxides by different peroxidases. Depending on the individual organism and the developmental state, the parasites also contain significant amounts of glutathione, mono-glutathionylspermidine and ovothiol, whereby all four low molecular mass thiols are directly (trypanothione and mono glutathionylspermidine) or indirectly (glutathione and ovothiol) maintained in the reduced state by trypanothione reductase. Thus the trypanothione system is central for any thiol regeneration and trypanothione reductase has been shown to be an essential enzyme in these parasites. The absence of this pathway from the mammalian host and the sensitivity of trypanosomatids toward oxidative stress render the enzymes of the trypanothione metabolism attractive target molecules for the rational development of new drugs against African sleeping sickness, Chagas' disease and the different forms of leishmaniasis. PMID- 12751785 TI - Glutathione--functions and metabolism in the malarial parasite Plasmodium falciparum. AB - When present as a trophozoite in human erythrocytes, the malarial parasite Plasmodium falciparum exhibits an intense glutathione metabolism. Glutathione plays a role not only in antioxidative defense and in maintaining the reducing environment of the cytosol. Many of the known glutathione-dependent processes are directly related to the specific lifestyle of the parasite. Reduced glutathione (GSH) supports rapid cell growth by providing electrons for deoxyribonucleotide synthesis and it takes part in detoxifying heme, a product of hemoglobin digestion. Free radicals generated in the parasite can be scavenged in reaction sequences involving the thiyl radical GS* as well as the thiolate GS-. As a substrate of glutathione S-transferase, glutathione is conjugated to non degradable compounds including antimalarial drugs. Furthermore, it is the coenzyme of the glyoxalase system which detoxifies methylglyoxal, a byproduct of the intense glycolysis taking place in the trophozoite. Proteins involved in GSH dependent processes include glutathione reductase, glutaredoxins, glyoxalase I and II, glutathione S-transferases, and thioredoxins. These proteins, as well as the ATP-dependent enzymes of glutathione synthesis, are studied as factors in the pathophysiology of malaria but also as potential drug targets. Methylene blue, an inhibitor of the structurally known P. falciparum glutathione reductase, appears to be a promising antimalarial medication when given in combination with chloroquine. PMID- 12751786 TI - Oxidative stress caused by inactivation of glutathione peroxidase and adaptive responses. AB - Reactive oxygen species (ROS) are generated as by-products of cellular metabolism, primarily in the mitochondria. When the cellular production of ROS exceeds the cell's antioxidant capacity, cellular macromolecules such as lipids, proteins and DNA can be damaged. Because of this, 'oxidative stress' is thought to contribute to aging and pathogenesis of a variety of human diseases. However, in the last 10-15 years, a considerable body of evidence has accumulated that ROS serve as subcellular messengers, and play a role in gene regulation and signal transduction pathways, which may be involved in defensive mechanisms against oxidative stress. This review focuses on oxidative stress caused by the inactivation of glutathione peroxidase (GPx), a major peroxide scavenging enzyme. GPx is inactivated by a variety of physiological substances, including nitric oxide and carbonyl compounds in vitro and in cell culture. Decreased GPx activity has also been reported in tissues where oxidative stress occurs in several pathological animal models. The accumulation of increased levels of peroxide resulting from inactivation of GPx may act as a second messenger and regulate expression of anti-apoptotic genes and the GPx itself to protect against cell damage. These findings suggest that GPx undergoes inactivation under various conditions such as nitroxidative stress and glycoxidative stress, and that these changes are a common feature of various types of oxidative stress which may be associated with the modification of redox regulation and cellular function. PMID- 12751787 TI - Versatility of selenium catalysis in PHGPx unraveled by LC/ESI-MS/MS. AB - Phospholipid hydroperoxide glutathione peroxidase (PHGPx; EC 1.11.1.12), a broad spectrum thiol-dependent peroxidase, deserves renewed interest as a regulatory factor in various signaling cascades and as a structural protein in sperm cells. We present a first attempt to identify catalytic intermediates and derivatives of the selenoprotein by liquid chromatography coupled to electrospray tandem mass spectrometry (LC/ESI-MS/MS) and to explain observed specificities by molecular modeling. The ground state enzyme E proved to correspond to position 3-170 of the deduced porcine sequence with selenium being present as selenocysteine at position 46. The selenenic acid form, which is considered to be the first catalytic intermediate F formed by reaction with hydroperoxide, could not be identified. The second catalytic intermediate G was detected as Se glutathionylated enzyme. This intermediate is generated in the reverse reaction where the active site selenol interacts with glutathione disulfide (GSSG). According to molecular models, specific binding of reduced glutathione (GSH) and of GSSG is inter alia facilitated by electrostatic attraction of Lys-48 and Lys 125. Polymerization of PHGPx is obtained under oxidizing conditions in the absence of low molecular weight thiols. Analysis of MS spectra revealed that the process is due to a selective reaction of Sec-46 with Cys-148' resulting in linear polymers representing dead-end intermediates (G'). FT Docking of PHGPx molecules allowed reactions of Sec-46 with either Cys-66', Cys-107', Cys-168' or Cys-148', the latter option being most likely as judged by the number of proposed intermediates with reasonable hydrogen bonds, interaction energies and interface areas. We conclude that the same catalytic principles, depending on the conditions, can drive the diverse actions of PHGPx, i.e. hydroperoxide reduction, GSSG reduction, S-derivatization and self-incorporation into biological structures. PMID- 12751788 TI - Modulation of the chymotrypsin-like activity of the 20S proteasome by intracellular redox status: effects of glutathione peroxidase-1 overexpression and antioxidant drugs. AB - ATP- and ubiquitin-independent proteolysis by the 20S proteasome is responsible for the selective degradation of oxidized proteins. In vitro, the 20S proteasome shows an increased proteolytic activity toward oxidized polypeptides and the suc LLVY-MCA peptide specific for its chymotrypsin-like activity. We have analyzed the effect of the intracellular redox status on the chymotrypsin-like activity of the 20S proteasome in human T47D cells overexpressing the detoxifiant enzyme seleno-glutathione peroxidase-1 (GPx-1). We report a 30% decreased activity of the chymotrypsin-like activity in cells overexpressing GPx-1. This phenomenon correlated with a 2-fold increase in IkappaB alpha half-life, a protein whose basal turnover is 20S proteasome-dependent. Following exposure to H2O2, these cells showed a seleno-dependently decreased accumulation of intracellular reactive oxygen species and 20S proteasome chymotrypsin-like activity. Similar results were obtained in HeLa cells transiently overexpressing human GPx-1. Moreover, exposure of HeLa cells to antioxidant compounds reduced the proteasome 20S chymotrypsin-like activity. In contrast, no effects were observed when HeLa cell extracts used to determine proteasome activity were incubated with either reduced or oxidized glutathione. These results suggest that GPx-1 activity or pro reducing conditions can downregulate basal 20S proteasome activity. Hence, the intracellular redox status, probably through the level of oxidized proteins, is an important element that can either activate or down-regulate the 20S proteasome chymotrypsin-like activity in living cells. PMID- 12751789 TI - Microflora trigger colitis in mice deficient in selenium-dependent glutathione peroxidase and induce Gpx2 gene expression. AB - Selenium-dependent glutathione peroxidase isoenzymes-1 and -2 are the major glutathione-dependent H2O2-reducing activities in the epithelium of the mid- to lower gastrointestinal tract. The two isoenzymes protect mice against ileocolitis. We have found that luminal microflora are required for colitis to develop in mice deficient in GPX-1 and GPX-2 activity (GPX-DKO). Within 7 days of association with microflora, previously asymptomatic germ-free GPX-DKO mice developed severe acute colitis while their littermates with at least one wild type Gpx1 or Gpx2 gene remained virtually symptom-free. Microflora also affected Gpx2 gene expression. Gpx2, but not Gpx1, mRNA levels were elevated 4-5 fold in the ileum and colon in conventionally reared or microflora-associated adult mice compared with germ-free mice. Since the gastrointestinal tract microflora undergo major changes 2-3 weeks after birth, from relatively benign to a potentially stressful composition, we examined postnatal Gpx2 gene expression. The jejunal and ileal GPX-2 activity levels were low in two to three week-old mice and increased 5-7 fold during the next two weeks. GPX-2 activity levels were correlated with the mRNA levels. Colon Gpx2 mRNA levels held steady at about 50% of adult levels from 12-21 days of age but were several times higher than ileal levels. Our results suggest that ileal Gpx2 mRNA and GPX-2 activity levels are induced by luminal microflora. This response is consistent with a role for GPX as an anti-inflammatory activity. PMID- 12751790 TI - Recruitment of the interleukin-1 receptor (IL-1RI)-associated kinase IRAK to the IL-1RI is redox regulated. AB - Interleukin-1 signaling is initiated by recruitment of adapter proteins and kinases to the type I interleukin-1 receptor (IL-1RI). It is modulated by accompanying redox processes at various levels, such as (auto-) phosphorylation of the IL-1RI-associated kinase IRAK, the phosphorylation of IkappaB and translocation and transcriptional activity of NF-kappaB. Here we demonstrate that the thiol-modifying agents diamide, menadione, and phenylarsine oxide (PAO) block the recruitment of IRAK to the receptor without inhibiting kinase activity in the immunoprecipitated IL-1RI complex in the human epithelial cell line ECV304 and the murine T cell line EL-4. Inhibition of IRAK receptor association by menadione is reversible in a GSH-dependent manner, while the PAO effect proved to be irreversible. Phospholipid hydroperoxide glutathione peroxidase attenuates inhibition by menadione. Recruitment correlates with the presence of thiol groups in IRAK that were available for IAIT-labeling. We conclude that recruitment of IRAK to the IL-1RI is redox regulated by the glutathione system, a reduced status being a prerequisite for an appropiate IL-1 response. PMID- 12751791 TI - Kinetics and redox-sensitive oligomerisation reveal negative subunit cooperativity in tryparedoxin peroxidase of Trypanosoma brucei brucei. AB - Tryparedoxin peroxidases (TXNPx) are peroxiredoxin-type enzymes that detoxify hydroperoxides in trypanosomatids. Reduction equivalents are provided by trypanothione [T(SH)2] via tryparedoxin (TXN). The T(SH)2-dependent peroxidase system was reconstituted from TXNPx and TXN of T. brucei brucei (TbTXN-Px and TbTXN). TbTXNPx efficiently reduces organic hydroperoxides and is specifically reduced by TbTXN, less efficiently by thioredoxin, but not by glutathione (GSH) or T(SH)2. The kinetic pattern does not comply with a simple rate equation but suggests negative co-operativity of reaction centers. Gel permeation of oxidized TbTXNPx yields peaks corresponding to a decamer and higher aggregates. Electron microscopy shows regular ring structures in the decamer peak. Upon reduction, the rings tend to depolymerise forming open-chain oligomers. Co-oxidation of TbTXNPx with TbTXNC43S yields a dead-end intermediate mimicking the catalytic intermediate. Its size complies with a stoichiometry of one TXN per subunit of TXNPx. Electron microscopy of the intermediate displays pentangular structures that are compatible with a model of a decameric TbTXNPx ring with ten bound TbTXN molecules. The redox-dependent changes in shape and aggregation state, the kinetic pattern and molecular models support the view that, upon oxidation of a reaction center, other subunits adopt a conformation that has lower reactivity with the hydroperoxide. PMID- 12751792 TI - Testis-specific expression of the nuclear form of phospholipid hydroperoxide glutathione peroxidase (PHGPx). AB - The selenoprotein phospholipid hydroperoxide glutathione peroxidase (PHGPx) is present in at least three different isoforms in testis: as a cytosolic, as a mitochondrial, and as a nuclear protein. We have recently shown that a sperm nucleus-specific glutathione peroxidase (snGPx) is identical to the mitochondrial and cytosolic forms of PHGPx apart from its N-terminus. This arginine-rich N terminus of snGPx, reminiscent of protamines, is encoded by an alternative exon located in the first intron of the PHGPx gene and is responsible for nuclear localisation and chromatin binding of snGPx [Pfeifer et al., FASEB J. 15 (2001), pp. 1236-1238]. By using a combination of techniques including selective cloning of mRNA 5'-ends, RT-PCR, and S1 analyses, we provide evidence that the transcript encoding the nuclear form is generated by transcription initiation at an alternative promoter and not by alternative splicing. We show that the major transcription start region is located at -12 to -14 upstream of the AUG translation initiation site of the sperm nucleus-specific exon and lacks a TATA box. Two minor TATA-less transcription initiation sites are located at around -30 and -45. We have shown by in situ hybridisation that snGPx expression in testis, like protamine expression, is restricted to late stages of spermatogenesis whereas PHGPx expression is only found in spermatocytes and early spermatids. These findings have to be taken into account when studying either the differential regulation of PHGPx and snGPx expression in testis or the impact of putative mutations in snGPx on male fertility in man. PMID- 12751793 TI - Selective recognition of peptide sequences by glutathione transferases: a possible mechanism for modulation of cellular stress-induced signaling pathways. AB - Exogenous and endogenous agents including products generated by oxidative stress, chemotherapeutics and bacterial lipids, activate multiple cellular signaling pathways, resulting either in mitogenesis or in apoptosis. Glutathione transferases (GSTs) appear not only to be prominent catalysts of detoxication reactions, but also to play a pivotal role in signaling by interacting with multiple proteins in pathways induced by cellular stress. Using two peptide libraries (a 9-mer and a 15-mer) displayed on phage, novel GST-peptide interactions were identified using human GST A1-1, GST P1-1 and GST M2-2 as targets. The isolated peptides have high sequence similarity with proteins such as TRAF4-associated factor 1, G protein-coupled receptor MRGX3, tumor necrosis factor superfamily (member 9), and c-Jun N-terminal kinase 3. PMID- 12751794 TI - Biosynthesis of trypanothione in Trypanosoma brucei brucei. AB - Trypanothione [T(SH)2], the major redox mediator in pathogenic trypanosomatids, is synthetized stepwise by two distinct enzymes in Crithidia fasciculata, while in Trypanosoma cruzi a single enzyme catalyzes both steps. A full-length reading frame presumed to encode trypanothione synthetase (TryS) was obtained by PCR using DNA of T. brucei as template and primers based on fragments of putative TryS genes. The recombinant protein produced by E. coli Origami (DE3) was purified to homogeneity by chelate and ion exchange chromatography. The enzyme catalyzed both reactions of T(SH)2 biosynthesis. Thus, T(SH)2 synthesis appears to be similar in African (T. brucei) and New World (T. cruzi) trypanosomes but distinct from that of Crithidia. PMID- 12751795 TI - Transcriptional regulation of cytosol and membrane alanyl-aminopeptidase in human T cell subsets. AB - Aminopeptidase inhibitors strongly affect the proliferation and function of immune cells in man and animals and are promising agents for the pharmacological treatment of inflammatory or autoimmune diseases. Membrane alanyl-aminopeptidase (mAAP) has been considered as the major target of these anti-inflammatory aminopeptidase inhibitors. Recent evidence also points to a role of the cytosol alanyl-aminopeptidase (cAAP) in the immune response. In this study we used quantitative RT-PCR to determine the mRNA expression of both cAAP and mAAP in resting and activated peripheral T cells and also in CD4+, CD8+, Th1, Th2 and Treg (CD4+ CD25+) subpopulations. Both mAAP and cAAP mRNAs were expressed in all cell types investigated, and in response to activation their expression appeared to be upregulated in CD8+ cells, but downregulated in Treg cells. In CD4+ cells, mAAP and cAAP mRNAs were affected in opposite ways in response to activation. The cAAP-specific inhibitor, PAQ-22, did not affect either cAAP or mAAP expression in activated CD4+ or CD8+ cells, whereas in activated Treg cells it markedly upregulated the mRNA levels of both aminopeptidases. The non-discriminatory inhibitor, phebestin, significantly increased the amount of mAAP and cAAP mRNA in CD4+ and that of cAAP in Treg cells. PMID- 12751796 TI - Regulation of gene transcription by a constitutively active mutant of activating transcription factor 2 (ATF2). AB - Activating transcription factor 2 (ATF2) belongs to the family of basic region leucine zipper (bZIP) proteins that are characterized by the presence of a basic domain that functions as the DNA-binding domain and a leucine zipper domain that is required for dimerization. Together with bZIP proteins of the Fos and Jun families, ATF2 constitutes the AP-1 transcription factor complex. The biological activity of ATF2 is controlled by phosphorylation of two threonine residues within the N-terminal activation domain. Unphosphorylated ATF2 is trancriptionally silent, excluding simple overexpression studies to identify transcriptional targets of ATF2. We therefore decided to construct a constitutively active ATF2 mutant that would allow us to uncouple the investigation of transcriptional targets and biological functions of ATF2 from the variety of signaling pathways that lead to an activation of ATF2. We exchanged the phosphorylation-dependent activation domain of ATF2 with the constitutively active transcriptional activation domain of the transcription factor CREB2. In transient transfection experiments, this constitutively active ATF2 mutant stimulated c-jun, tumor necrosis factor alpha, and Fas ligand promoter activities. The transcriptional activity of the constitutively active ATF2 mutant could be impaired by dominant-negative forms of ATF2 or c-Jun, indicating that ATF2 and c-Jun utilize a similar dimerization code. In contrast, a dominant-negative CREB2 mutant did not impair ATF2-mediated transcriptional activation, suggesting that CREB2 exhibits a different dimerization specificity than ATF2 or c-Jun. PMID- 12751797 TI - Solvent isotope effect on the reaction catalysed by the pyruvate dehydrogenase complex from Escherichia coli. AB - The pyruvate dehydrogenase from Escherichia coli showed a primary kinetic isotope effect when its overall reaction or the partial reaction of the pyruvate dehydrogenase component were tested in deuterium oxide. The Michaelis constants for pyruvate were nearly unchanged, but the maximum velocities in water and deuterium oxide differed, their ratio being DV = 1.7 for the overall reaction and DV = 2.1 for the E1p reaction. The pH profile and, accordingly, the delta pK1 and delta pK2 values were shifted by 0.6 units to higher pL values. A linear proton inventory curve was obtained when varying the atom fractions of protons relative to deuterons from 100 to 0%. This is an indication for a single proton transfer. It is proposed that this relatively weak primary isotope effect may be caused by the protonation of the N1' nitrogen at the pyrimidine ring of the cofactor by an adjacent glutamate residue. The proton of its carboxylic group exchanges very fast with deuterons of the solvent. PMID- 12751798 TI - Selective induction of liver parenchymal cell heme oxygenase-1 in selenium deficient rats. AB - Liver heme oxygenase (HO) activity is higher in selenium-deficient rats than in control animals under basal conditions and is further increased in them, but not in controls, by phenobarbital treatment. In the present study we characterized liver HO induction by selenium deficiency using molecular methods. Severe selenium deficiency in rats caused a doubling of liver HO activity without affecting spleen, kidney, brain, or testis HO activities. HO-1 protein and mRNA were increased to accompany the increased HO activity, but HO-2 protein and mRNA were not increased. Fractionation of the liver into hepatocyte and Kupffer cell/endothelial cell fractions revealed that the increased HO activity resides in the hepatocyte fraction. Immunohistochemical localization of HO-1 protein confirms the induction of HO-1 taking place solely in hepatocytes and throughout the liver lobule. Phenobarbital treatment sharply increased HO-1 mRNA and protein expression in selenium-deficient liver and HO activity in hepatocytes, but had no effect in control liver or in the Kupffer cell/endothelial cell fraction of selenium-deficient liver. Electrophoretic mobility shift assays showed increased AP-1 binding activity, suggesting an involvement of this redox-sensitive transcription factor in the induction by phenobarbital of HO-1 in selenium deficiency. We speculate that selenium deficiency affects hepatic antioxidant selenoproteins, resulting in an up-regulation of HO-1. PMID- 12751799 TI - Vagal rebound during resolution of tearful crying among depressed and nondepressed individuals. AB - Respiratory sinus arrhythmia (RSA) is an index of the vagal control of heart rate that is associated with emotion regulatory capacity. To examine RSA in depressed and nondepressed participants in the context of an emotion-regulatory challenge, we presented a sad film to induce crying, a behavior associated with heightened parasympathetic activation. We predicted that nondepressed persons who cried would show elevations in RSA during the onset and the resolution of crying. By contrast, we predicted that depressed individuals who cried would fail to exhibit increased RSA over the course of their crying episodes. As hypothesized, nondepressed participants exhibited RSA increases that accompanied the resolution of tearful crying, consistent with a homeostatic function for crying, whereas depressed subjects who cried did not exhibit increased RSA. Results suggest that the physiological self-regulatory mechanisms invoked by crying are compromised in depression. PMID- 12751800 TI - Are fingers special? Evidence about movement preparation from event-related brain potentials. AB - Ulrich, Leuthold, and Sommer (1998) suggested that movement preparation at the level of the motor cortex, as indexed by the lateralized readiness potential (LRP), proceeds in a strongly hierarchical fashion, where parameters other than response hand are prepared only if all the movement parameters are known. These conclusions were based on an experiment where a precue provided information about response hand, direction of finger movement, and movement force. To assess the generality of these findings, we replaced the force parameter with response finger. LRP indicated that preparation of the required finger is possible even when preliminary information is incomplete. Therefore, movement preparation appears to follow different rules when anatomical relationships (hand and finger) are concerned as compared to functional parameters like movement direction. On the other hand, at a limb-unspecific level, as indicated by the contingent negative variation, we confirmed evidence for parallel programming of all movement parameters. PMID- 12751801 TI - Sensory gating impairment associated with schizophrenia persists into REM sleep. AB - Physiological measures of sensory gating are increasingly used to study biological factors associated with attentional dysfunction in psychiatric and neurologic patient populations. The present study was designed to assess sensory gating during rapid eye movement (REM) sleep in patients with schizophrenia, a population bearing a genetic load for gating impairment. Auditory event-related potentials (ERPs) were recorded in response to paired clicks during separate waking and overnight sleep recording sessions in controls and schizophrenia patients. Suppression of ERP component P50 was significantly impaired in the patient group during both waking and REM sleep, whereas the difference between groups for N100 gating was dependent on state. These results suggest that REM sleep is an appropriate state during which to assess P50 gating in order to disentangle the effects of state and trait on sensory gating impairment in other clinical populations. PMID- 12751802 TI - Relationship between temporal changes in cardiac parasympathetic activity and motion sickness severity. AB - Chemotherapy-induced nausea has been associated with a time-related decrease in cardiac parasympathetic activity. We tested the hypothesis that a time-related decrease in cardiac parasympathetic activity would also be associated with nausea and other motion sickness symptoms during illusory self-motion (vection). Fifty nine participants (aged 18-34 years: 25 male) were exposed to a rotating optokinetic drum to induce vection. Symptoms of motion sickness and an estimate of cardiac parasympathetic activity (respiratory sinus arrhythmia; RSA) were obtained at baseline and throughout a drum-rotation period. As expected, motion sickness symptoms increased and RSA decreased over time during drum rotation. Moreover, greater decreases in RSA over time correlated with greater motion sickness severity. These results suggest that a time-related decrease in cardiac parasympathetic activity may be an important correlate of nausea and motion sickness across different evocative contexts. PMID- 12751803 TI - Event-related potential assessment of information processing after closed head injury. AB - We evaluated alterations in information processing after closed head injury as a function of task demands and stimulus modality. Visual and auditory discrimination tasks were administered to 11 survivors of a head injury and 16 matched healthy controls. In auditory tasks, compared with controls, the survivors had smaller N100s, smaller and later N200s, a more posterior scalp distribution of N200, and longer P300 and response latencies. Auditory N200 and P300 correlated highly with duration of unconsciousness. In contrast, in visual tasks, only a reduced N200 in the survivors differentiated the groups. Our results indicate that processing of auditory stimuli, including the perception and discrimination of stimulus features and the evaluation and categorization of stimuli, may be impaired after head trauma. Visual sensory processing may be spared, but higher-order visual processing involved in stimulus classification may be compromised. PMID- 12751804 TI - Auditory event-related potentials in humans and rats: effects of task manipulation. AB - The purpose of this study was to compare components of the rat and human auditory event-related potential (ERP) as generated in active oddball and passive single stimulus tasks. The rats were trained to discriminate between target and standard stimuli in an oddball task, whereas the human subjects received instructions. Task effects on various ERP components were found in both species. Interestingly, effects on the P3 component were similar in the species with regard to amplitude: Target stimuli elicited a higher amplitude in the oddball task than did standard stimuli. This might indicate that the P3 shares the same characteristics between species. However, the first four components occurred 1.82 times earlier in rats than in humans, expecting a P3 of about 200 ms in rats. The P3 in rats appeared at 380 ms. We conclude that either the relation between human and rat peak latencies is not linear, or the P3 in rats is not the equivalent of the human P3. PMID- 12751805 TI - Processes involved in tempo perception: a CNV analysis. AB - The purpose of this research was to study the mechanisms underlying tempo perception, by looking at their electrophysiological brain correlates. The subjects' task consisted of comparing the tempos of two isochronous tone sequences made up of either three (condition 13) or six (condition 16) 600-ms intervals. Contingent negative variation (CNV), known to be linked to the judgment of a single interval, kept increasing in amplitude for three intervals during tempo encoding, thereby providing evidence of the occurrence of CNVs also for several intervals in succession. This CNV increase could reflect the use of interval-based processes in the building of the interval memory trace. During the comparison phase, a CNV decrease was observed in condition 16, suggesting that subjects did not build a new memory trace, but used beat-based processes to check whether the beats of the new tempo occurred at the times they anticipated. PMID- 12751806 TI - The algorithmic complexity of multichannel EEGs is sensitive to changes in behavior. AB - Symbolic measures of complexity provide a quantitative characterization of the sequential structure of symbol sequences. Promising results from the application of these methods to the analysis of electroencephalographic (EEG) and event related brain potential (ERP) activity have been reported. Symbolic measures used thus far have two limitations, however. First, because the value of complexity increases with the length of the message, it is difficult to compare signals of different epoch lengths. Second, these symbolic measures do not generalize easily to the multichannel case. We address these issues in studies in which both single and multichannel EEGs were analyzed using measures of signal complexity and algorithmic redundancy, the latter being defined as a sequence-sensitive generalization of Shannon's redundancy. Using a binary partition of EEG activity about the median, redundancy was shown to be insensitive to the size of the data set while being sensitive to changes in the subject's behavioral state (eyes open vs. eyes closed). The covariance complexity, calculated from the singular value spectrum of a multichannel signal, was also found to be sensitive to changes in behavioral state. Statistical separations between the eyes open and eyes closed conditions were found to decrease following removal of the 8- to 12-Hz content in the EEG, but still remained statistically significant. Use of symbolic measures in multivariate signal classification is described. PMID- 12751807 TI - Energy resource depletion, task difficulty, and cardiovascular response to a mental arithmetic challenge. AB - This study examined further cardiovascular effects of energy resource depletion. Participants performed initially an easy counting task (Task A easy) or a hard counting task (Task A difficult) for 5 min. Shortly thereafter, they were provided the chance to earn a modest incentive by attaining a low performance standard (Task B easy) or a high performance standard (Task B difficult) on a mental arithmetic task. As expected, the Task A difficulty factor combined with the Task B difficulty factor to determine blood pressure responses during the second performance period. Whereas Task A easy participants evinced relatively stronger responses when Task B was difficult than when Task B was easy, Task A difficult participants evinced relatively stronger responses when Task B was easy than when Task B was difficult. These cardiovascular results partially replicate cardiovascular results from a prior depletion study using a cognitive, as opposed to a motor, challenge. They also extend the cardiovascular results from the prior study by demonstrating that the cardiovascular influence of energy depletion depends on the difficulty of the challenge with which people are confronted. PMID- 12751808 TI - Frontal EEG asymmetry and the behavioral activation and inhibition systems. AB - Two studies have examined whether there exists a relationship between resting frontal alpha asymmetry and the Behavioral Inhibition and Activation Scales (C. S. Carver & T. L. White, 1994), which are based on Gray's Behavioral Inhibition and Behavioral Activation Systems. Findings suggest that greater relative left frontal activity characterizes individuals higher in self-reported behavioral activation sensitivity (E. Harmon-Jones & J. J. B. Allen, 1997; S. K. Sutton & R. J. Davidson, 1997), and, in one instance, lower behavioral inhibition sensitivity (S. K. Sutton & R. J. Davidson, 1997). In the present study, relatively greater left frontal activity correlated positively with behavioral activation scores. No significant relationship between resting frontal alpha asymmetry and the behavioral inhibition score emerged. These data suggest that relatively greater left frontal activity is indeed an index of approach oriented, appetitive motivational tendencies, whereas the relationship between relative right frontal activity and the behavioral inhibition system is likely to be complex and not accounted for by behavioral withdrawal alone. PMID- 12751809 TI - Event-related potential indices of masked repetition priming. AB - Two experiments sought to identify event-related potential (ERP) correlates of masked repetition priming of words in lists and to verify that such effects are not due to brief prime durations. In Experiment 1, prime stimuli were masked and their durations were individually titrated for each participant. Targets that were immediate or delayed repetitions of masked primes resulted in attenuation of the N400, with little or no enhancement of a late positive component (LPC). Delayed, in-the-clear repetitions of unmasked targets led to attenuation of the N400 and enhancement of the LPC. Experiment 2 used similar stimulus timing parameters, but primes were unmasked. More typical unmasked repetition effects were observed for immediate repetitions including a larger attenuation of the N400 and enhancement of the LPC. These findings are discussed within current notions of the functional significance of the N400 and LPC. PMID- 12751810 TI - Affective modulation and prepulse inhibition of startle among undergraduates high and low in behavioral inhibition and approach. AB - Valence modulation and prepulse inhibition of startle were examined among 80 undergraduates scoring in the upper and lower quartiles of self-report measures of behavioral inhibition (BIS) and behavioral approach (BAS). Participants viewed a series of pleasant, neutral, and unpleasant pictures. Acoustic startle probes (102 dB) were presented during most pictures and during intertrial intervals, and a prepulse (120-ms SOA) preceded half of the probes. Valence modulation on no prepulse trials was greater among high-BAS than low-BAS participants. Consistent with theory regarding behavioral approach, post hoc tests demonstrated robust inhibition during pleasant versus neutral pictures among high-BAS participants, but not low-BAS participants. Valence modulation was reliable among high-BIS but not low-BIS participants, but the group difference was not significant. Contrary to our prediction, prepulse inhibition tended to be greater among high-BAS than low-BAS participants. The present data call attention to the role of individual differences in pleasant affective experience in startle modification. PMID- 12751811 TI - The effects of serial position and frequency of presentation of common stimulus features on orienting response reinstatement. AB - Two factors that might affect the novelty value of a test stimulus (the frequency of appearance of features common to the test stimulus and the set of preceding stimuli, and the serial position of these features) were systematically manipulated, and their effects on the electrodermal component of the orienting response (OR) were examined. We presented 256 participants with both verbal and pictorial stimulus sequences. Following 12 presentations of control stimuli, a test stimulus, which shared two common components with some of the control stimuli, was presented, followed by two additional presentations of control stimuli. The results revealed that recent presentations of the common components significantly reduced OR magnitude to the test stimulus, whereas the presentation frequency of common components had no significant effect. The implications of these findings for the feature-matching theory are discussed and a modification of the theory is proposed. PMID- 12751812 TI - ERP correlates of true and false recognition after different retention delays: stimulus- and response-related processes. AB - Performance and electrophysiological correlates of true and false recognition were examined after short (40 s) and long (80 s) delays. True recognition showed no significant decrease after a long delay, whereas false recognition increased. Early frontal and parietal ERP old/new effects, considered as correlates of familiarity and recollection, were observed across delay for true recognition. No frontal effect occurred in the long delay for false recognition. This absence may arise from weakened memory traces preventing familiarity discrimination for LUREs. Response-related analysis revealed an error-related negativity (ERN) for true and false recognition, assuming that the effect reflects at least partly an internal misrepresentation of the correct response. The larger and topographically different ERNs for false recognition suggest an additional contribution of increased task demands and conditions of high response uncertainty. PMID- 12751813 TI - Semantic integration in videos of real-world events: an electrophysiological investigation. AB - Event-related potentials (ERPs) discriminated between contextually appropriate and inappropriate objects appearing in video film clips of common activities. Incongruent objects elicited a larger negative-going deflection, which was similar to the N400 component described previously in association with words and static pictures and which has been argued to reflect the integration of semantic information into a mental representation of the preceding context. The onset of this potential occurred shortly after object presentation, indicating that semantic integration is a rapid online component of real-world perception. In addition, the anomalies in movies evoked a large late positive potential at posterior regions, suggesting that in event perception, semantic incongruity may trigger cognitive processes other than those mediating pure semantic integration. PMID- 12751814 TI - Kinetics of inhibition of polyphenol oxidase mediated browning in apple juice by beta-cyclodextrin and L-ascorbate-2-triphosphate. AB - Polyphenol Oxidase (PPO) mediated browning in raw fruits and vegetables is a major cause of quality deterioration in fruits and vegetables and derived food products. Here the rate of browning reaction in apple juice treated individually and in combination (1:1) of beta-Cyclodextrin (beta-CD) and L-Ascorbate-2 triphosphate (L-AATP) is described. It was observed that the rate of quinone formation can be minimized using a combination of beta-CD and L-AATP as compared to individual treatment with these agents. Kinetic experiments revealed that both compounds are non-competitive inhibitors of PPO. PMID- 12751815 TI - A comparative QSAR study on carbonic anhydrase and matrix metalloproteinase inhibition by sulfonylated amino acid hydroxamates. AB - A quantitative structure-activity relationship (QSAR) study is made on the inhibition of a few isozymes of carbonic anhydrase (CA) and some matrix metalloproteinases (MMPs), both zinc containing families of enzymes, by sulfonylated amino acid hydroxamates. For both enzymes, the inhibition potency of the hydroxamates is found to be well correlated with Kier's first-order valence molecular connectivity index 1chi(v) of the molecule and electrotopological state indices of some atoms. From the results, it is suggested that while hydroxamate CA binding may involve mostly polar interactions, hydroxamate-MMP and hydroxamate ChC (ChC: Clostridium histolyticum collagenase, another zinc enzyme related to MMPs) bindings may involve some hydrophobic interactions. Both MMPs and ChC also possess some electronic sites of exactly opposite nature to the corresponding sites in CAs. A group such as C6F5 present in the sulfonyl moiety is shown to be advantageous in both CA and MMP (also ChC) inhibitions, which is supposed to be due to the interaction of this group with Zn2+ ion present in the catalytic site of both families of enzymes. PMID- 12751816 TI - 3Beta-sulfamate derivatives of C19 and C21 steroids bearing a t-butylbenzyl or a benzyl group: synthesis and evaluation as non-estrogenic and non-androgenic steroid sulfatase inhibitors. AB - A series of C19 and C21 steroids bearing one or two inhibiting groups (3beta sulfamate and 17alpha- or 20(S)-t-butylbenzyl or benzyl) were synthesized and tested for inhibition of steroid sulfatase activity. When only a sulfamate group was added to dehydroepiandrosterone, androst-5-ene-3beta,17beta-diol, pregnenolone and 20-hydroxy-pregnenolone, no significant inhibition of steroid sulfatase occurred at concentrations of 0.3 and 3 microM. With only a t butylbenzyl or a benzyl group, a stronger steroid sulfatase inhibition was obtained in the androst-5-ene than in the pregn-5-ene series. Comparative results from the screening tests and the IC50 values have shown that the effect of a sulfamate moiety as a second inhibiting group can be combined to the t butylbenzyl or benzyl effect in the C19 and C21 steroid series. The 3beta sulfamoyloxy-17alpha-t-butylbenzyl-5-androsten-17beta-ol (10) was thus found to be the most active compound with IC50 values of 46 +/- 8 and 14 +/- 1 nM, respectively for the transformations of E1S to E1 and DHEAS to DHEA. The IC50 values of compound 10 are similar to that of 17alpha-t-butylbenzyl-estradiol, which was previously reported by our group as a good steroid sulfatase reversible inhibitor, but remains higher than that of the potent inactivators estrone-3-O sulfamate (EMATE) and 17alpha-t-butylbenzyl-EMATE. However, contrary to these two latter inhibitors, compound 10 did not induce any proliferative effect on estrogen-sensitive ZR-75-1 cells nor on androgen-sensitive Shionogi cells at concentrations tested, suggesting that this steroid sulfatase inhibitor is non estrogenic and non androgenic. PMID- 12751817 TI - Inhibition of retinoic acid metabolising enzymes by 2-(4-aminophenylmethyl)-6 hydroxy-3,4-dihydronaphthalen-1(2H)-one and related compounds. AB - In a search for inhibitors of all-trans retinoic acid (RA)-metabolising enzymes as potential agents for the treatment of skin conditions and cancer we have examined 2-(4-aminophenylmethyl)-6-hydroxy-3,4-dihydronaphthalen-1(2H)-one (5). Compound (5) is a moderate inhibitor of RA-metabolising enzymes in mammalian cadaverous tissue microsomes and homogenates as well as RA-induced enzymes in cultured human genital fibroblasts and HaCat cells. Overall (5) was more potent than or equipotent with ketoconazole, a standard inhibitor, in the cadaverous systems but less active towards the RA-induced cell culture systems. Examination of the data suggests that RA-induction generates metabolising enzymes not present in the cadaverous systems, which are more susceptible to inhibition by ketoconazole than (5). PMID- 12751818 TI - Inactivation of rabbit liver carbonyl reductase by phenylglyoxal and 2,3,4 trinitrobenzenesulfonate sodium. AB - The chemical modifications of rabbit liver carbonyl reductase (RLCR) with phenylglyoxal (PGO) and 2,3,4-trinitrobenzenesulfonate sodium (TNBS), which are respective chemical modifiers of arginine and lysine residues, were examined. RLCR was rapidly inactivated by these modifiers. Kinetic data for the inactivation demonstrated that each one of arginine and lysine residues is essential for catalytic activity of the enzyme. Furthermore, based on the protective effects of NADP+, NAD+ and their constituents against the inactivation of RLCR by PGO and TNBS, we propose the possibility that the functional arginine and lysine residues are located in the coenzyme-binding domain of RLCR and interact with the 2'-phosphate group of NADPH. PMID- 12751819 TI - Evaluation of 5-[1-(2-halo(or nitro)ethoxy-2-iodoethyl)]-2'-deoxyuridines as inhibitors of herpes simplex virus. AB - A series of 5-[1-(2-haloethyl(or nitro)ethoxy-2-iodoethyl)]-2'-deoxyuridines (3 7) and related uracil analogs (9-10) were prepared using 5-vinyl-2-deoxyuridine (2) and 5-vinyl uracil (8) as starting materials. The regiospecific reaction of 2 and 8 with iodine monochloride and an alcohol provided the target compounds 3-10. These analogs were evaluated in vitro for inhibitory activity against thymidine kinase (TK) positive and negative strains of herpes simplex virus type-1. The compounds 3-10 were either weak or non-inhibitory to HSV-1 replication. All compounds investigated exhibited low host cell cytotoxicity. PMID- 12751820 TI - The inhibition mechanism of guanidine hydrochloride on the catalytic activity of recombinant human protein disulfide isomerase. AB - Initial velocity enzyme kinetics was used to study the inhibition mechanism of guanidine hydrochloride (Gdm.Cl) on catalytic activity of recombinant human protein disulfide isomerase (rhPDI) in protein folding. Reduced C125A recombinant human interleukin 2 (C125A rhIL-2), the substrate, was dissolved in 8 M Gdm.Cl before it was diluted into the folding buffer to initiate the folding reactions. The final Gdm.Cl concentrations in the folding buffer were fixed at 0.2 M, 0.4 M, 0.6 M and 0.8 M. The reduced and native C125A rhIL-2 were resolved by reversed phase-high performance liquid chromatography (RP-HPLC). The simultaneous nonlinear fitting of the initial velocities of the native C125A rhIL-2 formation vs the reduced C125A rhIL-2 concentrations in the presence of different Gdm.Cl concentrations shows that the inhibition mechanism of Gdm.Cl on the catalytic activities of rhPDI is a mixed-type noncompetitive nonlinear inhibition. PMID- 12751821 TI - Acidic nonsteroidal anti-inflammatory drugs inhibit rat brain fatty acid amide hydrolase in a pH-dependent manner. AB - Previous studies have demonstrated that fatty acid amide hydrolase, the enzyme responsible for the metabolism of anandamide, is inhibited by the acidic non steroidal anti-inflammatory drug (NSAID) ibuprofen with a potency that increases as the assay pH is reduced. Here we show that (R)-, (S)- and (R,S)-flurbiprofen, indomethacin and niflumic acid show similar pH-dependent shifts in potency to that seen with ibuprofen. Thus, (S)-flurbiprofen inhibited 2 microM [3H]anandamide metabolism with IC50 values of 13 and 50 microM at assay pH values of 6 and 8, respectively. In contrast, the neutral compound celecoxib was a weak fatty acid amide hydrolase inhibitor and showed no pH dependency (IC50 values approximately 300 microM at both assay pH). The cyclooxygenase-2-selective inhibitors nimesulide and SC-58125 did not inhibit fatty acid amide hydrolase activity at either pH. The data are consistent with the conclusion that the non ionised forms of the acidic NSAIDs are responsible for the inhibition of fatty acid amide hydrolase. PMID- 12751822 TI - Effect of Rhus coriaria L. (Anacardiaceae) on superoxide radical scavenging and xanthine oxidase activity. AB - Rhus coriaria L. (Anacardiaceae) is a well-known spice in the Middle-East and grown in the Central and East Anatolia region of Turkey. A methanolic extract (water-soluble part constituents) of R. coriaria, was found to be an uncompetitive inhibitor of xanthine oxidase and scavenger of superoxide radical in vitro with IC50 values of 172.5 microg/mL and 232 microg/mL respectively. Superoxide radicals were generated either by an enzymatic or a non-enzymatic system, and scavenging ability was evaluated by the inhibition of nitroblue tetrazolium reduction. This study provides evidence that a crude extract of R. coriaria exhibits interesting antioxidant properties, expressed either by the capacity to scavenge superoxide radical or to uncompetitively inhibit xanthine oxidase. PMID- 12751823 TI - Biological evaluation of several coumarin derivatives designed as possible anti inflammatory/antioxidant agents. AB - Several linear and angular coumarins designed and synthesised as possible anti inflammatory and antioxidant agents were evaluated for their biological activities, using the carrageenin-induced rat paw oedema model. In general, the compounds were found to be potent anti-inflammatory agents. Compound (4) was found to possess protective properties in adjuvant-induced arthritis in rats. The compounds were found to interact with 1,1-diphenyl-2-picryl-hydrazyl stable free radical (DPPH) whereas most of them were essentially inactive in other tests. The anti-inflammatory activity seemed to be connected with their reducing activity. R(M) values were determined as an expression of their lipophilicity which was also calculated as clog P. Only a poor relationship existed between lipophilicity and anti-inflammatory activity. PMID- 12751824 TI - In vivo and in vitro effects of some plant hormones on rat erythrocyte carbonic anhydrase and glucose-6-phosphate dehydrogenase activities. AB - The present study was undertaken to determine in vivo and in vitro effects of some plant growth regulators on rat erythrocyte carbonic anhydrase (CA) and glucose-6-phosphate dehydrogenase (G6PD) activities. Both in vivo and in vitro, spermidine and kinetin did not affect enzymatic activities of CA and G6PD, whereas putrescine decreased these activities, and abscisic acid increased them. Since plants use such growth regulators, their effects should be considered on mammals consuming them since they may possess important biological effects. PMID- 12751825 TI - Artificial organs and transplantation. AB - Nowadays artificial devices are not able to totally and undefinitely replace the loss of function of all vital organs and artificial organs can be used only to bridge the time to transplantation, which must be considered the first choice in the therapeutical approach for many chronic diseases. Since general population aging process is leading to an increase of organ demand, the gap between performed and requested transplantation is hard to fill. Xenotransplantation is nowadays only an experimental alternative solution and we have to do our best using available artificial organs to increase and improve the survival of patients waiting for transplantation. In this meeting we particularly dealt about organ function replacing therapy, especially regarding the kidney, heart, liver, pancreas and ear. PMID- 12751826 TI - Clinical treatment of the aging process presently and immediate future trends. PMID- 12751827 TI - Bionic kidney. AB - Bionic Kidney is a project still in progress which aims at replacing all renal functions, which has been carried out in an ideal attempt to improve the overall results of Renal Replacement Therapy. It contains all the requisites for a complete rehabilitation from Uremia. As a futuristic mini-device implanted in the body, it should be a reliable support to Transplantation performance, considering the scarcity of kidney donors. PMID- 12751828 TI - Artificial kidney: status of the art and new perspectives. AB - Extracorporeal dialysis was first performed in 1943 and has become a routine for End Stage Renal Patients from the early sixties. In the last 30 years researchers have focused on biocompatibility of artificial materials and optimisation of removal of uremic toxins by the membrane as in the long term treatment many complications like amylodosis heart and bone lesions, accelerated amyloidosis and immune system failure can occur. From this point of view high flux dialytic membranes are currently considered more biocompatible therefore being able to prevent such diseases. PMID- 12751829 TI - Artificial heart: the moment and the future. PMID- 12751830 TI - Artificial liver support in the third millennium. AB - Analogous to the artificial kidney there is a need for an effective and safe liver support system to bridge patients with hepatic failure to liver transplantation or own liver regeneration. An overview is given of the biological and non-biological systems used in clinical practice in the past and at present. The conclusion is drawn that only the biological systems might have the potential to prolong life significantly in patients with acute liver failure. The systems with this potential are summarised. Both in Europe and the USA good bioreactors are available. Most of them are based on porcine hepatocytes, which have immunological and zoonotic drawbacks. What is missing is the well differentiated human hepatocyte in sufficient amounts. Successful development of this cell will be the crown on bioartificial liver research in the third millenium. PMID- 12751831 TI - The artificial pancreas. AB - In type 1 diabetes an absolute deficiency of insulin secretion requires exogenous insulin supply to guarantee the patient's life avoiding ketoacidotic coma and to prevent the chronic complications of diabetes. In order to obtain a more physiological replacement therapy different approaches have been pursued since the early 70s to create an artificial wearable pancreas able to deliver insulin according to the blood glucose values as determined by continuous monitoring. Four components are considered essential for the realisation of an artificial pancreas: the sampling system, the glucose sensor, the mathematical models and the related algorithms for the calculation of the insulin doses and the infusion system for the insulin delivery. At present the still unsolved issues are mainly represented by the availability of reliable continuous glucose monitor and control algorithms, while the new technologies allow for the miniaturisation of the system. PMID- 12751832 TI - Deafness and the bionic ear. PMID- 12751833 TI - Xenotransplantation: new perspectives for the third millennium. PMID- 12751834 TI - Advancement in dialytic technologies: where to go? PMID- 12751835 TI - Artificial cells for replacement of metabolic organ functions. AB - Artificial cells are being actively investigated for use in the replacement of cell and organ functions, especially related to metabolic functions. The earliest routine clinical use of artificial cells is in the form of coated activated charcoal for hemoperfusion. Implantation of encapsulated cells are being studied for the treatment of diabetes, liver failure, kidney failure and the use of encapsulated genetically engineered cells for gene therapy. Blood substitutes based on modified hemoglobin are already in Phase III clinical trials in patients with as much as 20 units infused into each patient during trauma surgery. Artificial cells containing enzymes are being developed for clinical trial in hereditary enzyme deficiency diseases and other diseases. Artificial cell is also being investigated for drug delivery and for other uses in biotechnology, chemical engineering and medicine. PMID- 12751836 TI - Artificial organs as a bridge to transplantation. AB - Current organ shortage is estimated to keep outpacing demand for years to come. Among the advocated strategies, artificial and bioartificial devices may prove beneficial to a wide category of patients on transplant waiting lists. Bionic organ science allows to reproduce organ architecture and function through a complex interplay of cellular and mechanical elements. Some bioartificial organs may well be used to replace anatomical defects, while others allow to compensate for failing organ functions and to bridge patients to transplantation. Among these latter, bioartificial liver (BAL) systems bear the highest potential for clinical application, even if their use is raising several controversial issues. These latter regard the identification and stratification of patients fit for transplantation, timing and type of transplantation after recovery, appropriateness of double-blind, randomized clinical trials and safety of animal and/or human cell lines. Nonetheless, bionic organ science needs to be regarded as a useful adjunct in the armamentarium of organ replacement therapies for the third millennium. PMID- 12751837 TI - New perspectives in hemodialytic strategies. PMID- 12751838 TI - Best quality treatment for patient well being: a way to reduce social costs and alleviate personal suffering in dialysis. PMID- 12751839 TI - Vitamin E-bonded cellulose membrane, lipoperoxidation, and anemia in hemodialysis patients. AB - In hemodialysis patients, oxidative stress results from an imbalance between the production of reactive oxygen species and antioxidant defense mechanisms. Recently, a new dialysis multi-layer membrane has been developed, by modifying the inner surface of regenerated cellulose to support a vitamin E coating. The aim of our study was to investigate the effects of hemodialysis treatment with vitamin E-modified membrane on anemia and erythropoietin requirement in a group of chronic uremic patients. Ten uremic, non diabetic, patients on standard bicarbonate dialysis were treated with vitamin E-bonded dialysis membrane for 12 months. Hematological parameters, erythropoietin requirement, serum vitamin E and serum malonyldialdehyde (MDA) were evaluated before starting the study and monthly. No significant changes in hemoglobin level, RBC count, hematocrit and EPO requirement were observed. Basal vitamin E levels were in the normal range (13.0 +/- 2.88 mg/L vs. 14.79 +/- 3.12 mg/L; NS). On the contrary, basal MDA levels were higher than those observed in the control group (1.87 +/- 0.36 vs. 1.13 +/- 0.18 mmol/mL; p < 0.01) and a significant decrease of MDA levels was found after 1 month of Excebrane treatment (1.39 +/- 0.25 nmol/mL; p < 0.02). In conclusion, the role of the "oxidative hemolysis" in the pathogenesis of anemia in CHD patients is still not clearly defined, but it could be of minor clinical relevance. Although the effectiveness of vitamin E-coated membranes as a scavenger of ROS allows a better control of intradialytic oxidative stress, it doesn't seem to contribute to clinical management of anemia in these patients. PMID- 12751840 TI - A triad of costimulatory molecules synergize to amplify T-cell activation in both vector-based and vector-infected dendritic cell vaccines. AB - The activation of a T cell has been shown to require two signals via molecules present on professional antigen presenting cells: signal 1, via a peptide/MHC complex, and signal 2, via a costimulatory molecule. Here, the role of three costimulatory molecules in the activation of T cells was examined. Poxvirus (vaccinia and avipox) vectors were employed because of their ability to efficiently express multiple genes. Murine cells provided with signal 1 and infected with either recombinant vaccinia or avipox vectors containing a TRIad of COstimulatory Molecules (B7-1/ICAM-1/LFA-3, designated TRICOM) induced the activation of T cells to a far greater extent than cells infected with vectors expressing any one or two costimulatory molecules. Despite this T-cell "hyperstimulation" using TRICOM vectors, no evidence of apoptosis above that seen using the B7-1 vector was observed. Results employing the TRICOM vectors were most dramatic under conditions of either low levels of first signal or low stimulator cell to T-cell ratios. Experiments employing a four-gene construct also showed that TRICOM recombinants could enhance antigen-specific T-cell responses in vivo. These studies thus demonstrate the ability of vectors to introduce three costimulatory molecules into cells, thereby activating both CD4+ and CD8+ T-cell populations to levels greater than those achieved with the use of only one or two costimulatory molecules. This new threshold of T-cell activation has broad implications in vaccine design and development. Dendritic cells infected with TRICOM vectors were found to greatly enhance naive T-cell activation, and peptide-specific T-cell stimulation. In vivo, peptide-pulsed DCs infected with TRICOM vectors induced cytotoxic T lymphocyte activity markedly and significantly greater than peptide-pulsed DCs. PMID- 12751841 TI - Peer and teacher ratings of third- and fourth-grade children's social behavior as a function of early maternal employment. AB - BACKGROUND: One of the more controversial issues related to maternal employment in the United States concerns the timing of entry into the workforce and its effect on children, particularly during the first year of the child's life. Some studies show deleterious effects on children, such as increases in aggression and noncompliance, while others document few negative and even positive effects of early employment. METHODS: This study examined the long-term effects of maternal employment during the child's first year of life on the social behavior of 171 third- and fourth-grade children in two-parent families. The moderating effects of child gender and social class were investigated. The extent to which stability in alternative care arrangements statistically explained links between early maternal employment and child outcomes was tested. RESULTS: After controlling for child gender, and maternal ethnicity, social class, and current employment status, third- and fourth-grade children whose mothers were employed during their first year of life evinced more acting out and less frustration tolerance and were nominated more often by peers for 'hitting' and 'being mean' than children whose mothers were not employed. There was some evidence that these associations were moderated by child gender and social class: boys, but not girls, whose mothers were employed during the first year were subsequently rated by teachers as acting out more than other children, and were also more likely to be nominated by peers for hitting. Higher nominations for hitting were only found in the working class. Finally, there was partial evidence that the number of alternative child-care arrangements during the first year accounted for the links between early maternal employment and subsequent child outcomes. CONCLUSIONS: These results are congruent with extant research that posits a risk of early employment on socioemotional development, but show that this risk is partially attributable to child-care instability. PMID- 12751842 TI - Parenting and the development of conduct disorder and hyperactive symptoms in childhood: a prospective longitudinal study from 2 months to 8 years. AB - BACKGROUND: This study investigated the early processes involved in the development of symptoms of conduct disorder and hyperactivity. METHOD: The study employed a prospective design, over a period from 2 months to 8 years. Detailed observational data of early and later mother-child interactions were collected, infant prefrontal function (the A not B task) was assessed, and symptoms of child conduct disorder and hyperactivity were rated by maternal report at age 5 and 8 years. RESULTS: The principal findings of the study were that emotional dysregulation on the A not B task at 9 months predicted symptoms of conduct disorder at 5 and 8 years, and delayed object reaching times on the same task predicted hyperactive symptoms at 5 years. These two developmental trajectories were associated with distinct patterns of early parenting that were strongly influenced by infant gender. Thus, in boys early emotional dysregulation was predicted by rejecting and coercive parenting, and delayed reaching on the A not B task by coercive parenting, whereas in girls only continuity from earlier infant behaviour could be demonstrated. There was strong continuity between these early infant behaviours and later child disturbance that was partially mediated by parenting for conduct disorder symptoms (maternal hostile parenting in boys, and maternal coercive parenting in girls), but not for hyperactive symptoms. CONCLUSIONS: These data would suggest that only in boys was there evidence for the existence of a sensitive period for the development of hyperactive symptoms, and to a lesser extent, conduct disorder symptoms. PMID- 12751843 TI - Predicting change in psychopathology in youth referred to mental health services in childhood or adolescence. AB - BACKGROUND: Little evidence is available on factors associated with persistence and change of psychopathology, and little is known about the predictive value of factors regarding change once problem behaviours exist. This study aims to evaluate change in level of scores of empirically derived problem patterns and to study factors that influence this change for children and adolescents referred to mental health services. METHOD: A referred sample (N = 1,652), aged 4 to 18 years at initial assessment, was followed up after a mean interval of 6.2 years. We used standardised information from parents, teachers and subjects, including the CBCL, YSR and TRF at both assessments. RESULTS: Subjects at follow-up scored significantly above the expected mean norm scores, although for most scores improvement was found. The strongest predicting factor for time 2 psychopathology was the corresponding time 1 score, odds ratios ranging from 1.6 to 21.7. Males and children older at intake improved more than females and younger children, respectively. CONCLUSIONS: Few child, family and treatment-related factors had additional predictive value over and above earlier psychopathology, and their contribution to the prediction of outcome was small. Findings indicate continuity of behavioural and emotional problems in clinically referred children and adolescents, and these problems should be viewed as chronic conditions. Girls referred for behavioural and emotional problems may form a group especially at risk for poor outcome. PMID- 12751844 TI - Predictors of outcome among high functioning children with autism and Asperger syndrome. AB - BACKGROUND: The objective of this paper is to assess the extent to which measures of cognitive abilities taken in an inception cohort of young high functioning children with autism and Asperger syndrome predict outcome roughly two and six years later. METHOD: Children who received a diagnosis of autism or Asperger syndrome (AS) and who had a nonverbal IQ score in the 'non-retarded' range were included in the inception cohort. Measures of language and nonverbal skills were taken when the children were 4-6 years of age and outcome assessments were completed when the children were 6-8 and 10-13 years of age. The three outcome measures consisted of scales of adaptive behaviours in socialisation and communication and a composite measure of autistic symptoms (abnormal language, abnormal body and object use, difficulties relating to others, sensory issues and social and self-help difficulties). RESULTS: The explanatory power of the predictor variables was greater for communication and social skills than for autistic symptoms. The power of prediction was stable over time but did differ by PDD subtype. In general, the association between language skills and outcome was stronger in the autism group than in the AS group. CONCLUSIONS: These results support the emphasis of early intervention programmes on language but more work needs to be done on understanding variables that influence outcome in social skills and autistic behaviours, particularly in those with AS. PMID- 12751845 TI - Holistic and part-based face recognition in children with autism. AB - BACKGROUND: There is substantial evidence that children with autism are impaired in face recognition. Although many researchers have suggested that this impairment derives from a failure of holistic face processing and a tendency to represent and encode faces on a part-by-part basis, this hypothesis has not been tested directly. METHOD: Holistic face processing was assessed by comparing children's ability to recognize a face part (eyes, nose, or mouth) in the context of the whole face in which it was learned with their ability to recognize the same face part in isolation. RESULTS: In Study 1, as expected, typically developing 9-year-olds (n = 27) and 11-year-olds (n = 30) were significantly better at recognizing face parts presented in the whole than in the part test condition, and this effect was limited to upright faces and not found for inverted faces. Consistent with prior findings, typically developing children were most accurate when face recognition depended on the eyes. In Study 2, high functioning children with autism (n = 22) evidenced a whole-test advantage for mouths only, and were markedly deficient when face recognition depended on the eyes. Their pattern of performance diverged from age- and IQ-matched comparison participants (n = 20), who performed similarly to the typically developing children in Study 1. CONCLUSIONS: These findings suggest that face recognition abnormalities in autism are not fully explained by an impairment of holistic face processing, and that there is an unusual significance accorded to the mouth region when children with autism process information from people's faces. PMID- 12751846 TI - Pitch memory, labelling and disembedding in autism. AB - BACKGROUND: Autistic musical savants invariably possess absolute pitch ability and are able to disembed individual musical tones from chords. Enhanced pitch discrimination and memory has been found in non-savant individuals with autism who also show superior performance on visual disembedding tasks. These experiments investigate the extent that enhanced disembedding ability will be found within the musical domain in autism. METHOD: High-functioning children with autism, together with age- and intelligence-matched controls, participated in three experiments testing pitch memory, labelling and chord disembedding. RESULTS: The findings from experiment 1 showed enhanced pitch memory and labelling in the autism group. In experiment 2, when subjects were pre-exposed to labelled individual tones, superior chord segmentation was also found. However, in experiment 3, when disembedding performance was less reliant on pitch memory, no group differences emerged and the children with autism, like controls, perceived musical chords holistically. CONCLUSION: These findings indicate that pitch memory and labelling is superior in autism and can facilitate performance on musical disembedding tasks. However, when task performance does not rely on long-term pitch memory, autistic children, like controls, succumb to the Gestalt qualities of chords. PMID- 12751847 TI - Inhibitory mechanisms in autism spectrum disorders: typical selective inhibition of location versus facilitated perceptual processing. AB - BACKGROUND: This study examined the inhibitory control mechanisms of selective attention in autism spectrum disorders. Two issues were engaged: First, we extend previous findings of normal inhibition of distractor identity in autism by examining whether inhibition of spatial location is also spared. The second issue concerns the selectivity of inhibition. In non-clinical participants inhibition is selectively directed to the properties of the distractor that compete for the control of action; we examined whether individuals with autism also show normal selectivity of inhibition. METHOD: A negative priming task was used to examine selective spatial inhibition in participants with autism relative to matched non clinical controls. RESULTS: We discovered that inhibition of distractor spatial location is within normal limits in autism, as is the ability to selectively direct inhibition to task-relevant stimulus features. In addition, we unexpectedly found that the irrelevant perceptual feature of colour produced a facilitation effect in autism, which has not been observed previously in typical controls. CONCLUSIONS: Evidence of colour facilitation implicates more fluent, but presumably less adaptive, perceptual processes in autism. PMID- 12751848 TI - A new Stroop-like measure of inhibitory function development: typical developmental trends. AB - BACKGROUND: Difficulties with inhibition are increasingly regarded as central to pathological behavioural and learning disorders in childhood. However, few measures are available to assist in the assessment of young children's inhibitory competence. METHOD: A new, Stroop-like measure of inhibitory function is described which was designed to be appropriate for use with children from 3 years of age. One hundred and fifty-five school children aged between 3 and 16 years took part in a study to examine the developmental characteristics and behavioural correlates of task performance. RESULTS: The task appears to provide a robust measure of inhibitory function across the age range, 3 to 16 years. Furthermore, the animal-stroop task appears to identify those at risk of hyperactive symptomatology within a school-based sample. CONCLUSIONS: The clinical and developmental applications of this new measure are discussed. PMID- 12751849 TI - Word reading and reading-related skills in adolescents with Williams syndrome. AB - BACKGROUND: Individuals with Williams syndrome have good spontaneous language despite low levels of intelligence. This study explores the relationship between intelligence, word decoding and reading-related skills in 20 individuals with Williams syndrome. METHODS: In addition to the KBIT, the participants were administered standardized measures of reading, vocabulary, rapid naming, phonological skills and an experimental measure of rhyme judgement. RESULTS: There was wide variability in the reading achievement among the individuals with WS. While some participants were unable to recognize letters of the alphabet, others scored within the normal range of tests of single word reading and decoding. Reading scores were correlated with intelligence as measured on KBIT matrices but not with the vocabulary measures. Reading also correlated with phonological awareness tasks yet, surprisingly, not with rapid naming. CONCLUSION: It is suggested that in individuals with retardation, intelligence rather than language and language-related skills predict achievements in word reading. PMID- 12751850 TI - Psychosocial correlates of depressive symptoms among 12-14-year-old Norwegian adolescents. AB - BACKGROUND: The aim of the study was to examine the relationships between various psychosocial factors and depressive symptoms in early adolescence. METHODS: A representative sample of 2,465 12-14-year-old adolescents comprising 50.8% girls and 49.2% boys, with a mean age of 13.7 years, was recruited in two counties in Norway. The participation rate was 88.3%. Depressive symptoms were measured by the Mood and Feelings Questionnaire (MFQ). RESULTS: Correlations between the total sum of stressful events/daily hassles and the total sum of MFQ were moderately high, rs = .49 and rs = .53, respectively. Depressive symptoms were more strongly correlated with school-related stress among boys than girls, whereas the correlation between daily hassles and depressive symptoms was higher for girls than boys. The results of univariate analyses showed significantly higher mean total MFQ scores among adolescents not living with both natural parents, those who had moved more than twice and those with more than 3 siblings orhaving fewer than 2 close friends. Further, adolescents from Third World societies and adopted adolescents, those from lower SES groups, having unemployed parents or living in coastal areas had higher mean depressive symptom scores. The results of multiple regression analyses yielded the following six significant predictors of total MFQ scores in order of importance: Sum of daily hassles and sum of stressful life events, gender, number of friends, ethnicity and mother's employment status. Altogether, these variables accounted for 43% of the total variance in MFQ scores. CONCLUSIONS: It is concluded that these psychosocial predictors should be addressed when assessing depressive symptoms in early adolescence. The findings of the study are discussed in view of previous research in the field and their clinical significance. PMID- 12751851 TI - Do sibling and friend relationships share the same temperamental origins? A twin study. AB - BACKGROUND: Friendship and sibling relationships differ in that there is an element of choice in friendships, whilst the latter represent a fixed and given relationship. The present study set out to investigate the temperamental correlates of friendships and sibling relationships, as well as to examine their genetic and environmental components. METHOD: Two hundred and twelve same-sex twin-pairs (102 MZ and 110 DZ; 104 male pairs and 108 female pairs) aged between 12 and 15, together with their parents, comprised the sample. The twins themselves reported on both negative and positive aspects of the two relationships, and all family members provided reports of the twins' temperament (mothers, fathers, and adolescent self-reports). RESULTS: Analyses revealed that negative aspects of both the sibling relationship and friendship were related to emotionality, while the positive aspects of these relationships were associated with sociability and activity. Genetic influences and the shared environment accounted for variance seen in the sibling relationship. In addition, nonshared environment accounted for a moderate amount of the sibling relationship variability and explained most of the variance for the friendship components. Finally, bivariate analyses of the covariance between temperament and close relationships highlighted the importance of nonshared environmental influences during adolescence. CONCLUSION: Three themes emerged. First, congruent temperamental expressions were seen across these two relationships' contexts. Second, friendship and the sibling relationship have distinct etiological profiles, indicating the child-specific character of adolescent friendships versus the reciprocal nature of the sibling relationship. Finally, the common influences to temperament and relationship quality provided evidence for the validity of adolescents' unique, nonshared experiences. PMID- 12751852 TI - The validity of analyses testing the etiology of comorbidity between two disorders: a review of family studies. AB - BACKGROUND: Knowledge regarding the causes of comorbidity between two disorders has a significant impact on research regarding the classification, treatment, and etiology of the disorders. Two main analytic methods have been used to test alternative explanations for the causes of comorbidity in family studies: biometric model fitting and family prevalence analyses. Unfortunately, the conclusions of family studies using these two methods have been conflicting. In the present study, we examined the validity of family prevalence analyses in testing alternative comorbidity models. METHOD: We reviewed 42 family studies that used family prevalence analyses to test three comorbidity models: the alternate forms model, the correlated liabilities model, or the three independent disorders model. We conducted the analyses used in these studies on datasets simulated under the assumptions of 13 alternative comorbidity models including the three models tested most often in the literature. RESULTS: Results suggest that some analyses may be valid tests of the alternate forms model (i.e., two disorders are alternate manifestations of a single liability), but that none of the analyses are valid tests of the correlated liabilities model (i.e., a significant correlation between the risk factors for the two disorders) or the three independent disorders model (i.e., the comorbid disorder is a third, independent disorder). CONCLUSION: Family studies using family prevalence analyses may have made incorrect conclusions regarding the etiology of comorbidity between disorders. PMID- 12751853 TI - The psychosocial functioning of children and spouses of adults with ADHD. AB - BACKGROUND: It is unclear what the impact of parental ADHD is on the day-to-day life of the rest of the family and how it contributes to the intergenerational transmission of this disorder. METHOD: The psychosocial functioning of 23 spouses and 63 children of 33 families with an ADHD parent and 20 spouses and 40 children of 26 comparison families was examined. Both adults and their spouses were assessed for lifetime and current Axis I and Axis II diagnoses, present general psychiatric symptoms and their marital relationships. Children were screened for ADHD and other problems, using the C-DISC, CBLC, TRF and the Social Adjustment Inventory. RESULTS: Children with an ADHD parent had higher rates of psychopathology than those from comparison families. Children with ADHD had more co-morbidities than non-ADHD children. Family and marital functions were impaired in ADHD families regardless of the gender of the affected parent. Children without ADHD from families with one psychiatrically healthy parent did well while the behaviour of children with ADHD was always poor and not associated with parental mental health. CONCLUSION: The results underscore the strong genetic contribution to ADHD and the need to carefully assess the non-ADHD parent as they seem to influence the well-being of non-ADHD children in families with an ADHD parent. PMID- 12751854 TI - Correlation between the diameter of the main renal artery and the presence of an accessory renal artery: sonographic and angiographic evaluation. AB - OBJECTIVE: To evaluate whether the diameter of the main renal artery is an indicator of the presence of an accessory renal artery in sonographic examination. METHODS: Of cases undergoing renal intraarterial digital subtraction angiography for any reason, those with no renal arterial disease detected on digital subtraction angiography were included in the investigation. Accessory renal arteries were disclosed by digital subtraction angiographic examinations. The dimensions of the kidney were measured sonographically. The lowest, highest, and mean values of diameters of main renal arteries were determined by sonography and angiography in both the group with accessory renal arteries and the group without accessory renal arteries. RESULT: Of kidneys with sonographically normal dimensions, angiographic evaluation was made in 107, and sonographic evaluation was made in 97, the renal arteries of which could be visualized optimally. Both radiologic methods showed that diameters of main renal arteries were significantly smaller in the presence of the accessory renal artery (P < .001, Student t test, Mann-Whitney U test, and receiver operating characteristic curve). CONCLUSIONS: In sonographic examination, the presence of the main renal artery with a diameter smaller than usual in a kidney with normal dimensions is indicative of the presence of an accessory renal artery. Taking this into account, we can obtain higher rates of detection of accessory renal arteries in sonographic examinations. PMID- 12751855 TI - Clinical application of sonographic elasticity imaging for aging of deep venous thrombosis: preliminary findings. AB - OBJECTIVE: Aging of deep venous thrombosis is an important and difficult clinical problem. Because it is known that thrombi harden as they mature, we have preliminarily tested sonographic elasticity imaging, a technique that estimates tissue hardness, to age venous thrombi. METHODS: Two adult patients with lower extremity thrombi were studied. One had a clinically chronic thrombus (at least 3 years old), whereas the other patient's thrombus was clinically subacute (25 days old). We performed freehand compression sonographic scans using a 5-MHz linear array transducer. Phase-sensitive B-scan frames were processed offline by a two dimensional complex correlation-based adaptive speckle-tracking technique. The distribution of internal strains in the wall of the vein, thrombus, and surrounding tissue was analyzed. Clot hardness was normalized to the venous wall. RESULTS: The chronic clot was homogeneous, and the strain in the chronic clot was at least 10 times smaller than that in the vessel wall. The subacute clot was much more heterogeneous, and, on average, the strain magnitude in the clot was 3 to 4 times greater than that in the vessel wall. CONCLUSIONS: In this preliminary work, the 2 thrombi appeared very different, and these results suggest that elasticity imaging may be able to age deep venous thrombosis. PMID- 12751856 TI - Doppler waveforms and blood flow parameters of the superior and inferior mesenteric arteries in patients having Behcet disease with and without gastrointestinal symptoms: preliminary data. AB - OBJECTIVE: To evaluate hemodynamic changes in mesenteric arteries in patients with Behcet disease with and without gastrointestinal symptoms. METHODS: Doppler sonography of mesenteric arteries was performed in 25 symptomatic and 15 asymptomatic patients having Behcet disease and in 25 healthy control subjects. The peak systolic, minimal, and mean velocities, resistive and pulsatility indexes, inner diameter, cross-sectional area, and blood flow volume of mesenteric arteries were evaluated. The results were compared between patient groups and controls. RESULTS: The mesenteric artery flow was significantly greater in patients in the symptomatic group than in those in the asymptomatic group or in controls. In the superior mesenteric artery, mean velocity and mean blood flow volume (0.35 +/- 0.18 m/s and 711 +/- 404 mL/min, respectively; P < .0001) in the symptomatic group were significantly higher than in the asymptomatic group (0.16 +/- 0.07 m/s and 305 +/- 168 mL/min, respectively) or in controls (0.15 +/- 0.07 m/s and 290 +/- 123 mL/min, respectively). The mean peak systolic velocity (1.23 +/- 0.47 m/s; P < .005) in the symptomatic group was significantly higher than in controls (0.93 +/- 0.23 m/s). In the inferior mesenteric artery, mean velocity and mean blood flow volume (0.25 +/- 0.10 m/s and 139 +/- 79 mL/min) in the symptomatic group were significantly higher than in the asymptomatic group (0.16 +/- 0.07 m/s; P < .006; and 78 +/- 26 mL/min; P < .007) or in controls (0.17 +/- 0.07 m/s; P < .003; 83 +/- 48 mL/min; P < .004). CONCLUSIONS: In this study, symptomatic patients with gastrointestinal Behcet disease were associated with a significant increase in mesenteric artery flow that could be evaluated easily on spectral patterns of arteries during Doppler sonography. The Doppler sonographic findings also revealed that intestinal involvement in patients with Behcet disease without gastrointestinal symptoms is not significantly different from that of healthy controls. PMID- 12751857 TI - Correlation between simultaneous scintigraphic and ultrasonographic measurement of gastric emptying in patients with type 1 diabetes mellitus. AB - OBJECTIVE: To compare scintigraphic measurements of total stomach emptying of a semisolid meal with ultrasonographic measurements of changes in antral area as estimates of antral emptying in type 1 diabetic patients. METHODS: Eleven patients with insulin-dependent diabetes mellitus were studied with simultaneous measurements of gastric emptying by scintigraphy and ultrasonography. Patients were imaged immediately after ingestion (time 0) and every 15 minutes over 120 minutes. The gastric emptying rate was expressed as percent reduction in antral cross-sectional area from 15 to 90 minutes after meal ingestion. RESULTS: Ultrasonographic measurements showed a postprandial maximal antral area at 15 minutes, continuously decreasing with time, and reaching a plateau 45 to 90 minutes after the end of meal ingestion, whereas the scintigraphic counts attained their maximum immediately after the meal and began to fall thereafter. Between 15 and 90 minutes, the residual radioactivity and antral ultrasonographically measured distension curves were concordant. The curves then showed a tendency toward deviation for the last 15 minutes (median, 51% versus 59% at 105 minutes and 40% versus 57% at 120 minutes, respectively). A strong significant correlation could be seen between the ultrasonographic gastric emptying rate and scintigraphic half-time values (r = -0.94; P < .001). Comparing scintigraphic and ultrasonographic half-time values showed a systematic measurement error of 9.9 minutes and a random measurement error of 18.6 minutes. CONCLUSIONS: The use of standardized real-time ultrasonography to determine the gastric emptying rate of semisolid meals in diabetic patients, with the use of the change in gastric antral cross-sectional area in a single section of the stomach 15 and 90 minutes postprandially, offers a valid method for clinical practice. PMID- 12751858 TI - Splenic trauma: evaluation with contrast-specific sonography and a second generation contrast medium: preliminary experience. AB - OBJECTIVE: To report our experience in the assessment of splenic trauma with contrast-coded sonography and a second-generation contrast medium. METHODS: From January to May 2002, 120 patients were studied with sonography for suspected splenic trauma. Twenty-five were selected for further imaging because of sonographic findings positive for splenic injury, findings positive for peritoneal fluid only, indeterminate findings, and negative findings with high clinical or laboratory suspicion. These patients underwent contrast-enhanced harmonic sonography and contrast-enhanced helical computed tomography. RESULTS: Among the 25 patients studied, 6 had no spleen trauma at initial and follow-up evaluation. One patient had a hypoperfused spleen without parenchymal damage, and 18 had splenic injuries; these 19 patients were considered positive. Hemoperitoneum was identified by sonography, contrast-enhanced sonography, and contrast-enhanced computed tomography in 74% of the 19 positive cases. Perisplenic clots were recognized in 58% of the cases by computed tomography and in 42% by baseline and enhanced sonography. Splenic infarctions were found in 11% of cases by contrast-enhanced sonography and computed tomography; none was found by unenhanced sonography. Parenchymal traumatic lesions were identified in 12 of 18 patients with splenic injuries by unenhanced sonography, in 17 cases by contrast-enhanced sonography, and in all 18 cases by contrast-enhanced computed tomography. A minimal splenic lesion was found in the single patient with a false negative contrast-enhanced sonographic finding. Contrast-enhanced sonography correlated appreciably better than unenhanced sonography in detecting injuries and in estimating their extent. Findings undetectable on unenhanced sonography were also noted: splenic hypoperfusion in 11% of positive cases on both contrast enhanced sonography and contrast-enhanced computed tomography, contrast medium pooling in 21% of cases on both contrast-enhanced sonography and computed tomography, and contrast extravasation in 11% of cases on computed tomography and 5% on contrast-enhanced sonography. CONCLUSIONS: Contrast-enhanced sonography is a promising tool in the assessment of splenic trauma. In institutions where sonography is used as the initial procedure, this technique may increase its effectiveness. PMID- 12751859 TI - Tissue harmonic imaging: utility in breast sonography. AB - OBJECTIVE: To determine the impact of tissue harmonic imaging on visualization of focal breast lesions and to compare gray scale contrast between focal breast lesions and fatty tissue of the breast between tissue harmonic imaging and fundamental frequency sonography. METHODS: A prospective study was performed on 219 female patients (254 lesions) undergoing sonographically guided fine-needle biopsy. The fundamental frequency and tissue harmonic images of all lesions were obtained on a scanner with a wideband 7.5-MHz linear probe. Twenty-three breast carcinomas, 6 suspect lesions, 9 fibroadenomas, 1 papilloma, 1 phyllodes tumor, 162 unspecified solid benign lesions, and 40 cysts were found. In 12 cases the fine-needle aspiration did not yield sufficient material. The gray scale intensity of the lesions and adjacent fatty tissue was measured with graphics software, and the gray scale contrast between lesions and adjacent fatty tissue was calculated. RESULTS: Tissue harmonic imaging improved the gray scale contrast between the fatty tissue and breast lesions in 230 lesions (90.6%; P < .001) compared with fundamental frequency images. The contrast improvement was bigger in breasts with predominantly fatty or mixed (fatty/glandular) composition than in predominantly glandular breasts. The overall conspicuity, lesion border definition, lesion content definition, and acoustic shadow conspicuity were improved or equal in the harmonic mode for all lesions. CONCLUSIONS: The tissue harmonic imaging technique used as an adjunct to conventional breast sonography may improve lesion detectability and characterization. PMID- 12751860 TI - Pregnancy- and lactation-associated breast cancer: mammographic and sonographic findings. AB - OBJECTIVE: To evaluate the mammographic and sonographic findings of pregnancy associated breast cancer. METHODS: A total of 22 consecutive patients with breast cancer pathologically diagnosed during pregnancy (n = 10) or lactation (n = 12) were included in this study. The ages of the patients ranged from 26 to 49 years. Both mammography and sonography were performed on 12 patients; sonography only was performed on 7 patients; and mammography only was performed on 3 patients. Mammographic and sonographic findings were evaluated retrospectively. RESULTS: Mammography revealed positive findings in 13 (86.7%) of 15 patients, even though all 15 patients had dense breasts. Mammographic findings included masses (n = 5), masses with calcifications (n = 2), calcifications with axillary lymphadenopathy (n = 2), a mass with axillary lymphadenopathy (n = 1), calcifications alone (n = 1), asymmetric density alone (n = 1), and diffuse skin and trabecular thickening alone (n = 1). Sonographic findings were positive and showed masses for all 19 patients (100%). The common sonographic findings of masses were irregular shapes (n = 15), irregular margins (n = 16), parallel orientation (n = 11), complex echo patterns (n = 14, including marked cystic [anechoic] components [n = 4]), and posterior acoustic enhancement (n = 12). Surrounding tissue effects could be seen in 5 patients, including ductal changes (n = 2), Cooper ligament thickening (n = 1), edema (n = 3), and skin thickening (n = 3). Calcifications within or outside a mass (n = 7) and axillary lymphadenopathy (n = 8) were also detected. CONCLUSIONS: Although a mass could not be discernible by mammography because of increased radiodensity during pregnancy or lactation, calcification, asymmetric density, axillary lymphadenopathy, and skin and trabecular thickening were helpful for diagnosis of pregnancy-associated breast cancer. Sonographic findings of a solid mass with posterior acoustic enhancement and a marked cystic component were somewhat different from the appearance of breast cancer in nonpregnant women, possibly because of the physiologic changes of pregnancy and lactation. PMID- 12751861 TI - Pregnancy outcome of embryonic/fetal pleural effusion in the first trimester. AB - OBJECTIVE: To investigate the incidence of embryonic/fetal pleural effusion in the first trimester and its pregnancy outcome. METHODS: A total of 965 viable singleton pregnancies confirmed by sonography between 7 and 10 weeks were examined to estimate the incidence of embryonic/fetal pleural effusion. When initial transvaginal sonography showed pleural effusion, serial ultrasound examinations were performed. RESULTS: Pleural effusion was detected in 12 pregnancies (incidence, 1.2%; 95% confidence interval, 0.7-2.2), which involved bilateral thoracic cavities in all cases. The pregnancy outcome was assessed among 14 cases of pleural effusion, including 2 previously reported cases from the same institution. Among these, 12 pregnancies (86%) miscarried by 14 weeks' gestation. Karyotype was abnormal in 9 (82%) of 11 cases in which chromosomal analysis was successfully performed. Of these, 6 (67%) were 45,X. CONCLUSIONS: The results suggested that embryonic/fetal pleural effusion in early pregnancy was associated with poor pregnancy outcome such as spontaneous abortion and chromosomal abnormality. PMID- 12751862 TI - Sonographically guided radio frequency thermal ablation for unresectable recurrent tumors in the retroperitoneum and the pelvis. AB - OBJECTIVE: To evaluate the feasibility of sonographically guided radio frequency thermal ablation as a minimally invasive method for treatment of unresectable recurrent or metastatic tumors in the retroperitoneum and the pelvis, which often pose difficult surgical problems. METHODS: Radio frequency thermal ablation was performed on 7 patients with unresectable recurrent retroperitoneal or pelvic tumors from colorectal (n = 4), renal (n = 2), and prostate (n = 1) cancers. Under sonographic guidance, a total of 11 radio frequency thermal ablation operations were performed by a percutaneous or transanal approach. RESULTS: Three patients were asymptomatic, whereas 4 patients were symptomatic. The sizes of the tumors ranged from 29 to 100 mm (mean, 50.5 mm). Radio frequency thermal ablation was technically completed in all operations without intraoperative complications. The ablation time ranged from 25 to 238 minutes depending on the tumor size. There was no mortality. There were postoperative complications in 3 operations (27.3%): an enterovesical fistula, a skin burn, and fecal incontinence. The hospital stay was generally 0 to 1 day. Tumor marker levels decreased after radio frequency thermal ablation in all operations. Symptoms of 4 patients were controlled by radio frequency thermal ablation. One patient with recurrent renal cancer and uncontrollable hypercalcemia became asymptomatic immediately after radio frequency thermal ablation. Local recurrence at the radio frequency thermal ablation site occurred in 2 patients (28.6%), but these local recurrent tumors were treated effectively by additional sonographically guided radio frequency thermal ablation. CONCLUSIONS: Minimally invasive sonographically guided radio frequency thermal ablation is technically feasible for local treatment of unresectable recurrent retroperitoneal and pelvic tumors from different origins. Care should be taken to avoid thermal injury to surrounding organs. Further study is needed to evaluate its safety and efficacy. PMID- 12751863 TI - Imaging of burned-out testis tumor: five new cases and review of the literature. AB - OBJECTIVE: Burned-out tumors of the testis are extremely rare. From 5 new cases, the clinical and radiologic findings are discussed in light of a review of the literature. METHODS: Over a 13-year period, 5 patients 17 to 50 years old were admitted with metastatic germ cell neoplasms. All were explored by thoracic and abdominal computed tomography and scrotal sonographic examination. RESULTS: The disease was revealed by the presence of lymphadenopathies in 4 of these patients: retroperitoneal in 3 and supraclavicular in the other. Scrotal sonography revealed abnormalities in all cases. CONCLUSIONS: Burned-out tumors may cause some confusion in the diagnosis, because secondary tumors can often be mistaken for primary tumors. Careful evaluation of the testis is crucial for identifying the primary lesion site; all abnormalities shown on clinical or sonographic scrotal examination call for orchidectomy. PMID- 12751864 TI - Huge cavernous hemangioma of the adrenal gland: sonographic, computed tomographic, and magnetic resonance imaging findings. AB - Cavernous hemangiomas of the adrenal gland are rare in clinical practice. Recently some cases have come to clinical attention as incidental findings on sonography, computed tomography (CT), or magnetic resonance imaging (MRI). With many reports regarding its CT or MRI manifestation, sonographic features of this entity have seldom been delineated. We report the case of a patient with a huge nonfunctioning cavernous hemangioma of the adrenal gland and describe the imaging findings of the lesion. PMID- 12751865 TI - Mass at the splenic hilum: a clue to torsion of a wandering spleen located in a normal left upper quadrant position. AB - Wandering spleen is an extremely rare condition in which the spleen is lacking its normal ligamentous attachments and therefore can move to an ectopic position in the abdomen or pelvis. Wandering spleen predisposes the patient to life threatening complications due to torsion of the spleen's vascular pedicle, with resulting splenic infarction, portal hypertension, and bleeding. Because of the nonspecific symptoms, imaging plays an important role. To our knowledge, only a few case reports describing wandering spleen in the pediatric population have been published, including 2 cases in infancy. We report a case of splenic torsion in an infant with a preoperative diagnosis made on the basis of color and power Doppler sonography. The diagnosis was confirmed by contrast-enhanced helical computed tomography (CT) and proved at surgery. On sonography, a diffusely hypoechoic spleen and a mass at the splenic hilum representing the torsed splenic pedicle were shown. The splenic hilar mass correlates with the CT "whirl" sign indicative of torsion. PMID- 12751866 TI - Presumptive intrahepatic congenital cystic adenomatoid malformation. AB - Congenital cystic adenomatoid malformation (CCAM) and extralobar pulmonary sequestration (EPS) are 2 rare types of bronchopulmonary malformations that typically appear in infancy and childhood. A CCAM is a hamartomatous lesion of the lung, thought to be the result of the cessation of bronchiolar maturation with overgrowth of mesenchymal elements. An EPS is a discrete mass of lung tissues separate from the tracheobronchial tree and is supplied by a systemic vascular source (usually aortic). As well, it forms outside the normal pleural investment. Conran and Stocker analyzed 50 cases of EPS and found that 50% (23 of 46) were associated with a coexistent CCAM. Rare cases of intra-abdominal EPS (IEPS) have been reported in the literature, and in these cases, CCAMs commonly occur within the pulmonary sequesterant. We present an interesting case of CCAM manifesting within the liver of a patient treated prenatally for a large pulmonary type II CCAM and review the reported cases of IEPS with CCAM. PMID- 12751867 TI - Morgagni hernia: a rare form of congenital diaphragmatic hernia. AB - A congenital diaphragmatic hernia (CDH) occurs in approximately 1 per 2000 pregnancies. Most CDHs that are recognized in utero are of the Bochdalek type, resulting from a posterior defect in the diaphragm. Morgagni hernias result from an anterior defect and are the least common type of CDH (5%). They are rarely seen in utero. Furthermore, when they are seen in utero, they may be mischaracterized. We present a case of an unusual type of Morgagni hernia diagnosed in utero. The type of Morgagni hernia reported here is particularly uncommon. Two cases with similar findings have been reported previously. The shared findings between these cases indicate that, although rare, this may be a recognizable type of CDH and one that is distinguishable from the Bochdalek type. PMID- 12751868 TI - Prenatal sonographic features of intralobar bronchopulmonary sequestration. AB - Congenital lung lesions that can be diagnosed on the basis of prenatal sonography include cystic adenomatoid malformations and extrapulmonary bronchopulmonary sequestrations. Intralobar bronchopulmonary sequestration is a rare congenital malformation of the lung. The prenatal sonographic features of this condition are reported here. PMID- 12751869 TI - Exposure to environmental tobacco smoke and urinary 1-hydroxypyrene levels in preschool children. AB - Environmental tobacco smoke (ETS) contains relatively high concentrations of polycyclic aromatic hydrocarbons (PAHs). Urinary 1-hydroxypyrene (1-OHP), a metabolite of pyrene, is a good indicator of PAH exposure in occupational studies. In this study, we investigated the relationship between urinary 1-OHP concentration and ETS exposure in preschool children. Forty preschool children, aged 24-76 months, were studied during November and December, 1999. Two spot urine specimens (one in the morning immediately after the subject woke up and the other at night before the subject went to bed) were collected 1 day after completion of a questionnaire, in order to determine 1-OHP concentrations by fluorescent spectrophotometry. Overall, urinary 1-OHP concentrations were relatively low but detectable (morning: median, 0.021 microg/g creatinine; range, 0.002-1.019 microg/g creatinine; night: median, 0.015 microg/g creatinine; range, 0.002-1.328 microg/g creatinine). Multiple linear regression analyses revealed that the total number of cigarettes smoked by the children's fathers during the 3 days prior to collection of the urine specimens was significantly associated with their urinary 1-OHP concentrations, after adjusting for other confounders. Each cigarette smoked by a child's father resulted in an average 9.6% increase in 1 OHP concentration in the morning urine specimen (95% confidence interval = 1.8 18.1%; p = 0.02). We did not find a significant increase in the 1-OHP concentration in night urine specimens (p = 0.19). Although the sample size was small, these findings indicate that urinary 1-OHP may be a suitable biomarker of ETS carcinogen exposure in children. PMID- 12751870 TI - Optic neuritis: clinical analysis of 27 cases. AB - We retrospectively reviewed 27 cases diagnosed as idiopathic optic neuritis between 1992 and 2001 at Kaohsiung Veterans General Hospital to assess the clinical features, visual prognosis, neuroimaging, laboratory studies, and development of multiple sclerosis in Chinese patients with optic neuritis. Patient age ranged from 13 to 54 years (mean, 35.8 +/- 11.3 years). Five cases presented as bilateral optic neuritis and 22 as unilateral. Visual function improved gradually from 2 weeks after treatment. Twelve (44.4%) cases showed disc swelling and ocular pain was also noted in 44.4% of patients. All cases that underwent visual field and visual evoked potential tests showed abnormality in lesion eyes. Of the 23 cases that underwent neuroimaging studies, including computerized tomography (17 patients) and magnetic resonance imaging (6 patients), 10 revealed optic nerve thickening. Four cases (14.8%) developed multiple sclerosis during follow-up (mean, 4.3 years). The incidence of disc swelling was higher than that reported by the Optic Neuritis Treatment Trial, but the incidence of initial ocular pain, the presence of periventricular plaques, and the development of multiple sclerosis were lower in our study. The unilateral group had significantly better visual outcome than the bilateral group. PMID- 12751871 TI - Analysis of emergency department utilization by elderly patients under National Health Insurance. AB - Elderly persons are thought to use emergency departments (EDs) disproportionately. This phenomenon has implications for policy-making as the population of the elderly continues to increase. This study aimed to characterize national ED utilization by elderly patients (> or = 65 years old), compared with that by younger patients (15-64 years old). The sample was selected from the National Health Insurance Research Database for the year 2000. There were 519,003 visits to adult EDs in the 12 medical centers sampled. The study used a secondary data analysis and included 14 variables. The elderly accounted for 28.5% of all adult ED visits. Compared with younger patients, a greater proportion of elderly patients were male (61.6% vs. 47.7%, p < 0.001), had chronic diseases (11.9% vs. 8.0%, p < 0.001), were major cases (6.7% vs. 4.1%, p < 0.001), made no co-payment (38.5% vs. 5.8%, p < 0.001), were frequent users of outpatient services (28.4% vs. 9.9%, p < 0.001), were higher-level emergencies (60.0% vs. 43.7%, p < 0.001), had longer stays in the ED (14.0% vs. 4.7%, p < 0.001), and had higher costs per visit (NT dollars 4,814 +/- 6,046 vs. 2,779 +/- 5,533, p < 0.001). In addition, elderly patients used 40.8% of total adult ED costs. Older patients have distinct patterns of ED use, and use emergency services at a higher rate than younger patients. ED staff should be knowledgeable about the unique and complex presentations of older ED patients. PMID- 12751872 TI - Effect of zirconia-modified magnesia investment on the casting of pure titanium. AB - Several investigations have examined magnesia-based investments for pure titanium casting. However, the thermal expansion value was insufficient at low casting temperatures and high interfacial reactivity occurred at high casting temperatures. The purpose of this investigation was to modify a magnesia-based investment by adding a heat-resistant mold material, zirconia, in different ratios to produce a more accurate titanium casting. The thermal expansion value was measured using a new precise automatic laser recording machine and pure titanium was cast using an argon casting machine. The marginal accuracy was measured using a stereomicroscope and the interfacial reactivity of the titanium was evaluated using X-ray diffraction analysis. The results indicate that adding different amounts of zirconia to a magnesia-based investment can increase its thermal expansion value and decrease the interfacial reactivity of the titanium. Maximal thermal expansion in the zirconia-modified investments significantly increased by 5-6 weight % (wt%) and peaked at 1.62% expansion. Selevest with 5 wt% zirconia had the smallest mean marginal discrepancy, 21.70 microm at 750 degrees C. Analysis of variance indicates significant differences in marginal discrepancy between zirconia-modified investments (p < 0.001). Adding zirconia can also decrease the interfacial reactivity of the titanium. The data indicated that proper amounts of zirconia (5-6 wt%) added to a magnesia-based investment can produce a more accurate and less interfacial reactive pure titanium casting. PMID- 12751873 TI - Retrocaval ureter: report of two cases and literature review. AB - Retrocaval ureter is a rare congenital anomaly in which the ureter passes behind, and is compressed by, the inferior vena cava. Its etiology is assumed to be abnormal embryologic development of the inferior vena cava as a result of atrophy failure of the right subcardinal vein in the lumbar portion. We report two cases of retrocaval ureter and review the relevant literature. One patient was a 7-year old boy who presented with right flank pain. The other was a 40-year-old male who was found to have right hydronephrosis accidentally on abdominal sonography during a health examination. Both underwent retrograde pyelography which showed the typical S-shape of a retrocaval ureter. Abdominal computerized tomography and magnetic resonance imaging confirmed the diagnosis of retrocaval ureter. Ureteroureterostomies were performed. One patient showed focal squamous metaplasia of the ureter and the other had chronic inflammation and fibrosis. Follow-up studies showed improvement in hydronephrosis and renal function in both patients. We conclude that retrocaval ureter is a rare disorder and surgical correction is usually effective. PMID- 12751874 TI - Emphysematous cystitis, a rare complication of urinary tract infection in a male diabetic patient: a case report. AB - Emphysematous cystitis is a rare complication of urinary tract infection, characterized by spontaneous gas formation in the urinary bladder due to bacterial fermentation. Approximately 50 to 80% of patients with this disease are diabetic, and there is a higher incidence in females. We report a case of emphysematous cystitis in a diabetic male who was admitted under the impressions of hypoglycemia, acute bronchitis, and chronic renal failure. Treatment of the emphysematous cystitis consisted of adequate urinary drainage, empirical antibiotic therapy, and strict blood sugar control. The patient recovered satisfactorily after 9 days of hospitalization. PMID- 12751875 TI - Paraganglioma of the urinary bladder: a case report. AB - Paragangliomas of the urinary bladder are very rare. To date, there are no reliable methods for predicting their clinical behavior, so long-term follow-up is required. We describe a paraganglioma of the urinary bladder in a 32-year-old male who presented with painless gross hematuria. Abdominal ultrasonography revealed a protruding mass with hematoma over the right lateral wall of the urinary bladder. The tumor was not completely resected by transurethral resection of bladder tumor (TURBT) due to intermingling with the bladder wall. Follow-up I131-metaiodobenzylguanidine was performed 3 weeks after surgery and suspected incomplete resection lesions were noted. Histologic examination of the tumor indicated paraganglioma of the urinary bladder. We also provide a brief review of the literature for comparison. PMID- 12751876 TI - Leiomyoma of the urinary bladder: a case report. AB - Leiomyoma of the urinary bladder is a rare tumor and constitutes 35% of benign mesenchymal bladder tumors. Herein, we report a case of leiomyoma of the bladder. A 32-year-old female was incidentally found to have an abnormal bladder mass on abdominal ultrasound during work-up for infertility. Bimanual examination then revealed a mass on the right side near the bladder neck area. On cystoscopic examination, an external compression mass covered with normal bladder mucosa could be seen at the right lateral wall. Computerized tomography revealed a homogeneous solitary tumor protruding into the urinary bladder from the right lateral wall without enlarged lymph nodes. Magnetic resonance imaging showed an intraluminal round mass at the right aspect of the urinary bladder, leading to suspicion of an intramural neurogenic or mesenchymal tumor. Surgical exploration was performed via a lower midline incision and a 5.0 x 4.5 x 2.2 cm mass was excised. The pathologic diagnosis was leiomyoma of the bladder. We discuss the diagnosis and management of leiomyoma of the bladder and briefly review the literature. PMID- 12751877 TI - Environmental interventions to enhance student adjustment: implications for prevention. AB - School settings, often the sites for the prevention of adverse outcomes or the promotion of adjustment, are usually not the actual targets of such interventions. However, some interventions focus on modifying the school or classroom environments themselves. This review examines such approaches, and considers how school regularities that might undermine student adjustment are addressed. The environmental interventions are clustered in terms of focus: on student-student interactions, on teacher and peer influences, and on organizational function and structure. Reasons for the paucity of environmental change efforts and the inherent difficulties are discussed, and recommendations for creating ways to undertake future environmental interventions in schools are offered. PMID- 12751878 TI - Individual and school level effects of perceived harm, perceived availability, and community size on marijuana use among 12th-grade students: a random effects model. AB - A hierarchical linear model was used to estimate the individual and school level effects for marijuana use among a national sample of 12th-grade students. School effects were small in comparison to individual level effects, accounting for 2.9% of the variance in marijuana use. At the individual level, perceived harm, perceived availability, and their interaction were significant predictors, each of which varied randomly across schools. Among two school-level predictors, the normative environment for perceived harm was not significant, but normative perceived availability predicted level of marijuana use. The effect of perceived availability on marijuana use was stronger in larger, compared to smaller communities. Results are discussed in light of the use of random regression methods for identifying school-specific patterns of risk and protection for prevention planning. PMID- 12751879 TI - Two studies examining environmental predictors of heavy drinking by college students. AB - Two sequential studies are presented that examine the validity of a set of environmental variables to predict heavy drinking at college students' most recent drinking occasions. Random telephone interviews (n=1609, n=400) of graduate and undergraduate students attending two large public universities in the southwestern United States were conducted during three separate surveys in 2000 and 2001. An original interview schedule was used and it included measures that examined environmental characteristics of students' most recent drinking events, motivations for drinking, demographics, and alcohol consumption. Using nonparametric exploratory and confirmatory discriminant analyses to distinguish between heavy episodic and non-heavy episodic drinking events, a discriminant function was identified that included the following environmental variables: (1) having many people intoxicated at an event, (2) having illicit drugs available at an event, (3) BYOB events and, (4) the playing of drinking games at the event. The validity of these environmental variables to predict heavy drinking among students was supported in a subsequent study examining a separate sample from the same student population. Environmental factors can be useful to predict heavy drinking events experienced by students. Prevention programs would benefit from targeting such factors in combination with more traditional individual-level approaches. PMID- 12751880 TI - Effects of a preventive intervention on adolescent substance use initiation, expectancies, and refusal intentions. AB - This study evaluated the effects of a school-based preventive intervention (Botvin, G. J. 1996, 2000) on growth trajectories of substance initiation (alcohol, tobacco, and marijuana), expectancies, and refusal intentions. A rural midwestern sample (N=847) provided three waves of data from middle school students. Growth curve analyses demonstrated that the intervention significantly slowed the rate of increase in substance initiation and significantly slowed the rate of decrease in refusal intentions. The intervention also slowed the rate of decrease in negative outcome expectancies, although the significance level was only marginal. A multiple group comparison showed that the impact of initial levels of substance initiation on growth trajectories of refusal intentions differed between conditions, suggesting that the intervention decreased the effect of early substance initiation on the rate of change over time for refusal intentions. Gender differences also were found, although the intervention was effective in slowing the rate of increase in initiation for both genders. PMID- 12751881 TI - Precursors, consequences and implications for stability and change in pre adolescent antisocial behaviors. AB - Although much of the evidence stresses the stability of dysfunctional behavior throughout the life cycle, other evidence suggests that stability of antisocial behavior is a matter of degree. In this work we determine the degree of stability of such behavior in preadolescence and how this is influenced by age, gender, social structures, and family processes. Also, we explore whether change in the level of antisocial behavior impacts upon other important developmental regimes such as health and educational performance. We use a large, 2 wave, nationally representative sample of preadolescent children, and focus on children 4-9 years of age at wave 1 (n=6,846). We employ a cluster analysis across a series of behavioral variables to determine levels of antisocial behavior and then examine the stability of antisocial behavior over time and identify the precursors and consequences associated with movement into and out of these behavioral clusters. Antisocial behavior is more stable in boys and older children. Structural factors -age of the mother, number of children in the household, and having a single parent--along with family factors--hostile parenting and maternal depression- raise the likelihood of increases in and lower the likelihood of decreases in antisocial behavior, although there are notable differences by gender of the child and initial level of antisocial behavior. Consequences of change in antisocial behavior include scholastic performance, high levels of school mobility, school-parent contacts, and health perceptions. The implications of these findings for prevention and intervention programs are discussed. PMID- 12751882 TI - Nursing knowledge and the problem of worldviews. AB - Over the past few decades, nurse scholars have widely embraced the idea of worldviews and of worldviews serving as the bases of conceptions of nursing. Today, worldviews are so much taken for granted as the starting point of nursing inquiry that nurse scholars are continually seeking solutions to epistemological problems within the idea of worldviews. This article argues that, as long as we persist in doing so, the nursing discipline's aim to develop an organized body of nursing knowledge for nursing practice will remain unrealized. I propose that conceptions of nursing and of nursing inquiry be grounded in the philosophy of moderate realism, which eschews the idea of worldviews. PMID- 12751883 TI - Concept advancement: enhancing inductive validity. AB - Enhancing inductive validity while advancing a concept has always been a dilemma for qualitative researchers. This dilemma is paramount for researchers completing a series of small discrete projects all focused on the same concept. How do researchers use knowledge from their earlier research while maintaining an inductive paradigm? Template comparison, a method of sequential triangulation, is proposed as a solution to this problem. Template comparison allows the researcher to use findings from earlier studies, which employed the same or different qualitative methods, as a guide for the data collection, interpretative, and theoretical phases of the new study. The authors' research of the concept of trust is used as an example of the application of template comparison. PMID- 12751884 TI - The Delirium Observation Screening Scale: a screening instrument for delirium. AB - The Delirium Observation Screening (DOS) scale, a 25-item scale, was developed to facilitate early recognition of delirium, according to the Diagnostic and Statistical Manual-IV criteria, based on nurses' observations during regular care. The scale was tested for content validity by a group of seven experts in the field of delirium. Internal consistency, predictive validity, and concurrent and construct validity were tested in two prospective studies with high risk groups of patients: geriatric medicine patients and elderly hip fracture patients. Among the patients admitted to a geriatric department (N = 82), 4 became delirious; among the elderly hip fracture patients (N = 92), 18 became delirious. The DOS scale was determined to be content valid and showed high internal consistency, alpha = 0.93 and alpha = 0.96. Predictive validity against the Diagnostic and Statistical Manual-IV diagnosis of delirium made by a geriatrician was good in both studies. Correlations of the DOS scale with the Mini Mental State Examination (MMSE) were Rs -0.79 (p < or = 0.001) in the hip fracture patients and Rs -0.66 (p < or = 0.001) in the geriatric medicine patients. Concurrent validity, as tested by comparison of the research nurse's ratings of the DOS scale and the Confusion Assessment Method (CAM), for the group of hip fracture patients was 0.63 (p < or = 0.001). Construct validity of the DOS was tested against the Informant Questionnaire of Cognitive Decline in Elderly (IQCODE), a preexisting psychiatric diagnosis and the Barthel Index. Correlation with the IQCODE was 0.74 (p < or = 0.001) in the study with the hip fracture patients and 0.33 (p < or = 0.05) in the study with the geriatric medicine patients. Correlation with the Barthel Index was -0.26 (p < or = 0.05) in the geriatric medicine patients and -0.55 (p < or = 0.001) in the hip fracture patients. The overall conclusion of these studies is that the DOS scale shows satisfactory validity and reliability, to guide early recognition of delirium by nurses' observation. PMID- 12751885 TI - Partnership in practice. AB - Modern health care has increasingly focused on prescriptive, outcomes-oriented, and cost-effective practices concomitantly obscuring the humanness of the health experience. A reconsideration of partnership between nurse and client as the core of the discipline might call nurses back to what is essential to nursing: a caring relationship centered on that which is meaningful as health. This article points to the significance of the relational nature of partnership, differentiating its features and form from the prevalent understanding associated with prescriptive interventions to achieve predetermined goals and outcomes. The meaning of partnership is presented as nursing practice as it unfolds: a process of nurse and client relationship through which the caring presence of the nurse becomes integral to the health experience of the client as the potential for action. Exemplars provide illustration of this emerging view in practice and research. The article is intended to contribute to the expanding dialogue on nursing practice, inviting discussion of the relevance of partnership in different health systems. PMID- 12751886 TI - Meaning of dependency on care as narrated by 10 patients. AB - This article is part of an ongoing study that aims to illuminate the meaning of dependency on care. The aim of this particular study is to disclose the meaning of dependency on care as narrated by patients. We conducted interviews with patients (six men and four women) who had been in medical or surgical wards for at least 14 days. Seven of the patients were also interviewed one week after discharge. The participants ranged in age from 41 to 84 years old. The interviews were tape-recorded and transcribed verbatim. A phenomenological-hermeneutic approach was used to interpret the resulting text. The results show that to be dependent on care is to face the inevitability of not being able to manage by oneself--it is being attached to the nurses and bound to the care they offer. Being dependent on care involves a struggle to get care without treading on the nurses' toes. The nurses are one's lifelines and getting care is essential, no matter what. It is better to receive any form of care, good or bad, than to receive nothing. Being dependent on care is to be exposed and subjected to a nurse's ability and benevolence. One comprehensive understanding of the meaning of dependency on care is simply that "one does not saw off the branch one is sitting on." Furthermore, dependency on care involves a struggle to move forward in a life that hurts. Patients lose much of their freedom of choice in daily life and grieve their loss of ability and value. Patients may be able, however, to see things they would have never noticed earlier in life. Dependency on care is understood as one kind of "limit situation." Patients who are dependent on care may reevaluate their potential in life and gain another perspective on life. PMID- 12751887 TI - Nursing information. PMID- 12751888 TI - Practice environments. PMID- 12751889 TI - The mammary gland and its origin during synapsid evolution. AB - Lactation appears to be an ancient reproductive trait that predates the origin of mammals. The synapsid branch of the amniote tree that separated from other taxa in the Pennsylvanian (>310 million years ago) evolved a glandular rather than scaled integument. Repeated radiations of synapsids produced a gradual accrual of mammalian features. The mammary gland apparently derives from an ancestral apocrine-like gland that was associated with hair follicles. This association is retained by monotreme mammary glands and is evident as vestigial mammary hair during early ontogenetic development of marsupials. The dense cluster of mammo pilo-sebaceous units that open onto a nipple-less mammary patch in monotremes may reflect a structure that evolved to provide moisture and other constituents to permeable eggs. Mammary patch secretions were coopted to provide nutrients to hatchlings, but some constituents including lactose may have been secreted by ancestral apocrine-like glands in early synapsids. Advanced Triassic therapsids, such as cynodonts, almost certainly secreted complex, nutrient-rich milk, allowing a progressive decline in egg size and an increasingly altricial state of the young at hatching. This is indicated by the very small body size, presence of epipubic bones, and limited tooth replacement in advanced cynodonts and early mammaliaforms. Nipples that arose from the mammary patch rendered mammary hairs obsolete, while placental structures have allowed lactation to be truncated in living eutherians. PMID- 12751890 TI - The origin of lactation as a water source for parchment-shelled eggs. AB - Available evidence indicates that mammary gland secretions first evolved in synapsids that laid parchment-shelled eggs. Unlike the rigid-shelled eggs of birds and some other sauropsids, parchment-shelled eggs lose water very rapidly when exposed to ambient air of lower vapor pressure, whether due to differences in relative humidity or to differences in temperature. This precludes endothermic incubation of parchment-shelled eggs in an open nest. Synapsids may have avoided egg desiccation by incubating eggs in a pouch, but this would limit maternal activity. Parchment-shelled eggs are able to take up liquid water across the eggshell. I propose that mammary secretion originally evolved as a means of supplying water to eggs, and as such was essential to the evolution of endothermy among the egg-laying cynodonts that were ancestral to mammals. It is possible that synapsid eggs, like parchment-shelled squamate eggs, were also capable of uptake of some nutrients, such as sodium and ionic calcium. Living monotremes still produce parchment-shelled eggs. The porous eggshell and bilaminar yolk sac membrane of these eggs permit substantial uptake of uterine secretions during the intrauterine period, and might also facilitate uptake of mammary secretions during egg incubation. In its simplest form, mammary secretion may be an ancient trait of egg-laying synapsids, having had an important role long before milk became obligatory for suckling young. PMID- 12751891 TI - The evolutionary origins of maternal calcium and bone metabolism during lactation. AB - Calcium is required for skeletal growth in all vertebrate offspring. In eutherian mammals, calcium is provided by the mother via the placenta during fetal growth and via milk until weaning. Transferring calcium to offspring during pregnancy and lactation significantly stresses maternal calcium homeostasis. During human pregnancy, the extra calcium requirements are met primarily by an increase in absorption of calcium from the diet and by a modest increase in rates of bone resorption. In nursing mothers, the calcium required for milk production is generated by a dramatic increase in rates of bone resorption and a decrease in the rate of renal calcium excretion. To consider the evolution of these maternal adaptations in bone and calcium metabolism, comparisons are made across different species of mammals, and the fundamental problem of maternal transfer of calcium to young is explored in lower vertebrates. These comparisons suggest that maternal adaptations in calcium and bone metabolism during pregnancy and lactation in mammals originate from adaptations in bone and mineral metabolism that supply calcium for egg production in lower vertebrates. PMID- 12751892 TI - Evolution of the mammary gland defense system and the ontogeny of the immune system. AB - A decisive event in the evolution of mammals from synapsid reptiles was the modification of ventral thoracic-abdominal epidermal glands to form the mammary gland. The natural selection events that drove the process may have been the provision of certain immunological agents in dermal secretions of those nascent mammals. This is mirrored by similar innate immune factors in mammalian sebum and in protherian and eutherian milks. On the basis of studies of existing mammalian orders, it is evident that immune agents in milk such as immunoglobulins, iron binding proteins, lysozyme, oligosaccharides, and leukocytes compensate for developmental delays in early postnatal production of antimicrobial factors. At least in human milk, anti-inflammatory and immunomodulating agents also evolved to provide different types of protection for the offspring. In addition, investigations reveal that the types or concentrations of immunological agents in milk vary depending upon the type of placenta, lactation pattern, and environment of the species. PMID- 12751893 TI - Growth hormone and prolactin--molecular and functional evolution. AB - Growth hormone, prolactin, the fish hormone, somatolactin, and related mammalian placental hormones, including placental lactogen, form a family of polypeptide hormones that share a common tertiary structure. They produce their biological effects by interacting with and dimerizing specific single transmembrane-domain receptors. The receptors belong to a superfamily of cytokine receptors with no intrinsic tyrosine kinase, which use the Jak-Stat cascade as a major signalling pathway. Hormones and receptors are thought to have arisen as a result of gene duplication and subsequent divergence early in vertebrate evolution. Mammalian growth hormone and prolactin show a slow basal evolutionary rate of change, but with episodes of accelerated evolution. These occurred for growth hormone during the evolution of the primates and artiodactyls and for prolactin in lineages leading to rodents, elephants, ruminants, and man. Placental lactogen has probably evolved independently on three occasions, from prolactin in rodents and ruminants and from growth hormone in man. Receptor sequences also show variable rates of evolution, corresponding partly, but not completely, with changes in the ligand. A principal biological role of growth hormone, the control of postnatal growth, has remained quite consistent throughout vertebrate evolution and is largely mediated by insulin-like growth factors. Prolactin has many and diverse roles. In relation to lactation, the relative roles of growth hormone and prolactin vary between species. Correlation between the molecular and functional evolution of these hormones is very incomplete, and it is likely that many important functional adaptations involved changes in regulatory elements, for example, altering tissue of origin or posttranscriptional processing, rather than change of the structures of the proteins themselves. PMID- 12751894 TI - The comparative biology of whey proteins. AB - Lactational strategies and associated development of the young have been studied in a diverse range of species, and comparative analysis allows common trends and differences to be revealed. The whey fraction contains a vast number of proteins, many of which have not been assigned a function. However, it is expected that an understanding of the comparative biology of these proteins may provide some promise in assigning a function to the major whey proteins. Whey acidic protein is a major component of the whey fraction that has been studied across a range of species, revealing conservation of gene structure, whereas regulation and temporal expression patterns vary. This review focuses primarily on comparative analysis of whey acidic protein, highlighting gene structure, developmental and hormonal regulation, and potential functional roles for this protein. In addition, the contrasting regulation and secretion profiles of several other major whey proteins are discussed. PMID- 12751895 TI - Multispecies comparison of the casein gene loci and evolution of casein gene family. AB - Caseins, the major milk proteins, are present in a genomic cluster spanning 250 350 kb. The divergence at the coding level between human, rodent, and cattle sequences is rather extensive for most of the genes in this region. Nevertheless, comparative analysis of genomic sequences harboring the casein gene cluster region of these species (with equal evolutionary distances 79-88 Myr) shows that the organization and orientation of the genes is highly conserved. The conserved gene structure indicates that the molecular diversity of the casein genes is achieved through variable use of exons in different species and high evolutionary divergence. Comparative analysis also revealed the presence within two species of uncharacterized casein family members and ruled out the previously held notion that another gene family, located in this region, is primate-specific. Several other new genes as well as conserved noncoding sequences with potential regulatory functions were identified. All genes identified in this region are, or are predicted to be, secreted proteins involved in mineral homeostasis, nutrition, and/or host defense, and are mostly expressed in the mammary and/or salivary glands. These observations suggest a possible common ancestry for the genes in this region. PMID- 12751896 TI - The mammary gland in mammalian evolution: a brief commentary on some of the concepts. AB - Current thinking is highlighting the mammary glands and the process of lactation in the evolutionary success of mammals over and above the selective advantages provided by the nutritional and antimicrobial properties of milk. The extended period of contact between mothers and their young, necessitated by the regular and frequent transfer of milk, particularly characteristic of the primate strategy of reproduction and the primate mode of life, has been suggested to afford the offspring the opportunity for more learning and the eventual development of the levels of intelligence present in "higher" primates. Lactation offers the opportunity for maternal effects on development and the eventual phenotype of the offspring in addition to those that occur during pregnancy or from behavioral interactions. Lactation comes with high metabolic costs, which are manifested in parent-offspring conflict, and special physiological adaptations have evolved which match milk supply to demand by the young. PMID- 12751897 TI - On the mechanisms of post-rest adaptation in the isolated electrically driven left atria of rats. AB - We studied the role of the resting period (1, 2, 4, 8, 16 min; n = 6-7), external Ca2+ (0.2, 0.4, 0.6 g/l; n + 5-6), stimulation frequency (1, 2, 3 Hz; n = 6), 4 aminopyridine (4-AP, 2 mM; n = 5); theophylline (1 mM; n = 6), ouabain (5 microM; n = 6), and verapamil (1 microM; n = 6) on post-rest adaptation in the isolated left atria of rats driven electrically by a 2x threshold intensity for 2 ms. Resting periods resulted in three-phasic adaptive changes in contractility during the post-rest stimulation before normalization: P1, hypercontractile phase, an initial twitch potentiation; P2, post-rest hypocontractile decay reached after 8 to 12 single twitches; and P3, a late reactive hypercontractile phase marked less than that of P1 and gradually declining to the pre-resting level. P1 and P2 were augmented along with increasing the resting period from 1 min to 16 min, whereas t1 (time between P1 and P2) shortened and P2 and t2 (time between P2 and P3) were not affected. P1 and P3 to become more apparent after shifting the stimulation frequency from 1 Hz to 3 Hz, accompanied by a shortening of t1 and t2 (p < 0.05) and an insignificant reversal of P2. An increase in Ca2+ concentration by 2- or 3 fold at 2 Hz reduced P1 was and antagonized P2, while leaving other parameters almost unaffected. The reduction of P1 by Ca2+ became more prominent at 3 Hz. Exposure to 4-AP depressed P1 and P3 at 1 Hz, which was reversed by increasing the stimulation frequency--P3 tended to diminish, whereas t1 and t2 were shortened. Theophylline reduced P1 antagonized P2, and shortened t1 and t2 significantly, and a combination of theophylline and 4-AP augmented the effects. Ouabain increased P1 and P2 in a frequency-dependent manner; prolonged t2 at 1 Hz, but shortened t2 at higher frequencies. Verapamil inhibition of Ca2+ channels augmented P1 and t1 and reduced P2 and P3, and the effects on all three parameters were augmented by combined 4-AP/verapamil. We concluded that the post rest adaptive changes in contractility are a consequence of phasic changes in sarcoplasmic Ca2+ concentration and that such changes reflect an imbalance between the release from and uptake into the sarcoplasmic reticulum of Ca2+ and transsarcolemmal Ca2+ loss. PMID- 12751898 TI - The interaction of carbachol and strophantin on the electrical and mechanical events in electrically driven strips of guinea pig left atria. AB - We compared the electrophysiological and the mechanical actions of the cholinergic agonist carbachol (CARB) and the digitalis glycoside strophantin (STR) in a study on the electrically driven strips of guinea pig left atria. The respective control values for the resting membrane potential (RMP), action potential amplitude (APA), action potential duration at 20%, 50%, and 90% repolarization levels (APD20,50,90), and time to peak tension (TtPt) were of the order of 61.1 +/- 1.69 nmV, 104.64 +/- 1.28 mV, 28.5 +/- 1.22 msec, 51.0 +/- 1.51 msec, 106.91 +/- 3.81 msec, and 60.1 +/- 1.09 msec. Exposure to CARB (2 x 10(-7) M) rapidly reduced contractility, TtPt, APD, and APA to exceptionally lower levels within the initial 5 minutes and gradually caused a slight hyperpolarization in RMP. Strophantin (1 x 10(-7) M) caused contractility and RMP to increase slightly, APD and APA to reduce less markedly, but did not significantly affect TtPt. Under the influence of combined CARB/STR, the effect of CARB on the contraction amplitude (CA), TtPt, APD, and APA was attenuated and that on RMP was significantly potentiated (p < 0.05), whereas the effects of STR on CA, TtPt, APD, and APA were reversed (p < 0.05), and the effect on RMP was augmented. The action potential duration was less responsive to STR at all depolarization levels with respect to those of CARB and combined CARB/STR. Although the correlation between the time course of APD-20 and APD-90 for STR was poor, the correlation for CARB and combined CARB/STR was highly significant (r > 0.95). The correlation between the effects of CARB on CA and APD-90 was high but poor for the effect of combined CARB/STR. Striking to note is that the rate of induction of contractile changes with CARB (d(delta)/dt = 1.39 +/- 0.05%/sec), and its washout was more rapid (p < 0.05) in comparison to the rate of changes in APD-90 (0.98 +/- 0.05%/sec). This result, however, was not observed with combined CARB/STR. From the results we concluded that CARB and STR act as antagonists when used in combination, with the exception that only the effect on RMP was additive, with asignificant discordance between the rates of induction of effects on CA and APD-90, with the former being more rapid. PMID- 12751899 TI - Ultrastructural changes induced by leaves of Azadirachta indica (Neem) in the testis of albino rats. AB - The present work was designed to study the effect of Azadirachta indica (Neem) powder on rat testis using the electron microscope. Male albino rats received 100 mg each A. indica leaf powder orally (by gavage). On alternate days, a second group of rats received 0.125 mg testosterone dipropionate intramuscularly. A third group received both A. indica leaf powder by gavage and testosterone dipropionate intramuscularly. Suitable controls were maintained. After autopsy, ultrastructural analysis of the testis revealed that animals treated with testosterone dipropionate showed well-developed Sertoli cells and germ cells with well-developed cytoplasmic organelles. By contrast, in A. indica-treated rats, intracellular spaces and vacuolization were observed in Sertoli cells; whereas in Leydig cells, cytoplasmic inclusions appeared diminished, and the configuration of granular endoplasmic reticulum appeared as a single unbranched tubule. In late spermatids, defects were observed in the mitochondrial sheath. The ultrastructural changes seen in the A. indica-treated group provide a clue that A. indica leaves might affect spermatogenesis through antispermatogenic and antiandrogenic properties. PMID- 12751900 TI - The anti-implantation action of endosulfan in albino mice: possible mechanisms. AB - Endosulfan, a chlorinated pesticide, was administered orally for 7 consecutive days to a group of virgin pregnant albino mice at doses of 1 to 4 mg/kg/d to examine its effect on implantation. For comparison, a second group of animals received 5 microg/kg/d estradiol-17beta, a third group received similar quantities of olive oil. Autopsy on day 8 revealed that the olive oil-treated mice were pregnant with a normal number of implantations and a normal duration of diestrus. Treatment with estradiol-17beta completely inhibited implantation and significantly decreased the duration of diestrus with a concomitant increase in estrus. Treatment with 1, 2, or 3 mg/kg/d endosulfan neither inhibited implantation nor significantly changed diestrus, whereas 4 mg/kg/d endosulfan completely inhibited implantation, with the uterus showing no signs of implantation. This group exhibited a significant decrease in diestrus with a concomitant increase in estrus. In endosulfan-treated mice, no significant changes occurred in body and organ weight, except for a significant increase in uterine weight in groups treated with 4 mg/kg/d endosulfan or estradiol-17beta. Because endosulfan is neither tubal locking nor causes expulsion of the blastocyst from the uterus like estradiol-17beta, the pesticide might inhibit implantation by altering the estrogen-progesterone ratio essential for implantation. PMID- 12751901 TI - The peroxisomal membrane targeting elements of human peroxin 2 (PEX2). AB - Peroxin 2 (PEX2) is a 35-kDa integral peroxisomal membrane protein with two transmembrane regions and a zinc RING domain within its cytoplasmically exposed C terminus. Although its role in peroxisome biogenesis and function is poorly understood, it seems to be involved in peroxisomal matrix protein import. PEX2 is synthesized on free cytosolic ribosomes and is posttranslationally imported into the peroxisome membrane by specific targeting information. While a clear picture of the basic targeting mechanisms for peroxisomal matrix proteins has emerged over the past years, the targeting processes for peroxisomal membrane proteins are less well understood. We expressed various deletion constructs of PEX2 in fusion with the green fluorescent protein in COS-7 cells and determined their intracellular localization. We found that the minimum peroxisomal targeting signal of human PEX2 consists of an internal protein region of 30 amino acids (AA130 to AA159) and the first transmembrane domain, and that adding the second transmembrane domain increases targeting efficiency. Within the minimum targeting region we identified the motif "KX6(I/L)X(L/F/I)LK(L/F/I)" that includes important targeting information and is also present in the targeting regions of the 22-kDa peroxisomal membrane protein (PMP22) and the 70-kDa peroxisomal membrane protein (PMP70). Mutations in this targeting motif mislocalize PEX2 to the cytosol. In contrast, the second transmembrane domain does not seem to have specific peroxisomal membrane targeting information. Replacing the second transmembrane domain of human PEX2 with the transmembrane domain of human cytochrome c oxidase subunit IV does not alter PEX2 peroxisome targeting function and efficiency. PMID- 12751902 TI - Heterogeneity of microvascular endothelial cells isolated from human term placenta and macrovascular umbilical vein endothelial cells. AB - The present study compares some phenotypic and physiologic characteristics of microvascular and macrovascular endothelial cells from within one human organ. To this end microvascular endothelial cells from human full-term placenta (PLEC) were isolated using a new method and compared with macrovascular human umbilical vein endothelial cells (HUVEC) and an SV40-transformed placental venous endothelial cell line (HPEC-A2). PLEC were isolated by enzymatic perfusion of small placental vessels, purified on a density gradient and cultured subsequently. Histological sections of the enzyme-treated vessels showed a selective removal of the endothelial lining in the perfused placental cotyledons. The endothelial identity of the cells was confirmed by staining with the endothelial markers anti-von Willebrand factor, Ulex europaeus lectin and anti QBEND10. The cells internalized acetylated low-density lipoprotein and did not show immunoreactivity with markers for macrophages, smooth muscle cells and fibroblasts. The spindle-shaped PLEC grew in swirling patterns similar to that described for venous placental endothelial cells. However, scanning electron microscopic examination clearly showed that PLEC remained elongated at the confluent state, in contrast to the more polygonal phenotype of HPEC-A2 and HUVEC that were studied in parallel. The amount of vasoactive substances (endothelin 1,2, thromboxane, angiotensin II, prostacyclin) released into the culture medium and the proliferative response to cytokines was more similar to human dermal microvessels (MIEC) derived from non-fetal tissue than to HUVEC. Potent mitogens such as vascular endothelial growth factors (VEGF121, VEGF165) and basic fibroblast growth factor (FGF-2) induced proliferation of all endothelial cell types. Placental growth factors PIGF-1 and PIGF-2 effectively stimulated cell proliferation on PLEC (142 +/- 7% and 173 +/- 10%) and MIEC (160 +/- 20% and 143 +/- 28%) in contrast to HUVEC (9 +/- 8% and 15 +/- 20%) and HPEC-A2 (15 +/- 7% and 24 +/- 6%) after 48 h incubation time under serum-free conditions. These data support evidence for (1) the microvascular identity of the isolated PLEC described in this study, and (2) the phenotypic and physiologic heterogeneity of micro- and macrovascular endothelial cells within one human organ. PMID- 12751903 TI - Osteogenic potential of rat spleen stromal cells. AB - Evidence is mounting that an increasing number of cell populations in the adult organism already committed and/or differentiated retain the ability to reprogram themselves and give rise to a different phenotype. Bone marrow stromal cells have long been recognized as early progenitor cells for osteoblasts, chondrocytes, hematopoietic-supportive fibroblasts and adipocytes. Recent reports though have demonstrated a potential of cell populations outside the bone marrow environment to sustain bone formation under specific circumstances. The formation of bone nodules in the spleen of IL-5 transgenic mice has been recently reported (Macias et al. (2001): J. Clin. Invest. 107, 949 - 959). We thus postulated that a cell population exists in the spleen that under particular microenvironmental conditions is able to reprogram itself and pursue a fate other than the tissue specific one. Therefore we isolated and expanded in vitro spleen-derived stromal cells. After expansion, these cells were challenged with culture conditions designed to induce osteogenic differentiation. We hypothesized that the combination of a proliferating factor (fibroblast growth factor 2) and a differentiating hormone (dexamethasone) would allow us to induce spleen-derived stromal cells to proliferate and at the same time to express osteoblast-specific genes. Thus, spleen-derived stromal cells were isolated from rat spleen and expanded in the presence of fibroblast growth factor 2 and dexamethasone. Once primary cultures reached confluence they were either switched to an osteo inductive medium or implanted in immunodeficient mice. Although no bone formation was observed in in vivo experiments, in vitro spleen-derived stromal cells were able to deposit a mineralized matrix. Gene expression, as revealed by RT-PCR analysis, evidenced that the deposition of a mineralized matrix was concomitant with the expression of CBFA1 and osteocalcin, along with alkaline phosphatase and bone sialoprotein. Our data suggest that rat spleen-derived stromal cells can undergo osteogenic differentiation in a permissive microenvironment. PMID- 12751904 TI - Effect of stretching on gene expression of beta1 integrin and focal adhesion kinase and on chondrogenesis through cell-extracellular matrix interactions. AB - Differentiation of skeletal tissues, such as bone, ligament and cartilage, is regulated by complex interaction between genetic and epigenetic factors. In the present study, we attempted to elucidate the possible role of cell-extracellular matrix (ECM) adhesion on the inhibitory regulation in chondrogenesis responding to the tension force. The midpalatal suture cartilages in rats were expanded by orthopedic force. In situ hybridization for type I and II collagens, immunohistochemical analysis for fibronectin, alpha5 and beta1 integrins, paxillin, and vinculin, and cytochemical staining for actin were used to demonstrate the phenotypic change of chondrocytes. Immunohistochemical analysis for phosphorylation and nuclear translocation of extracellular signal-regulated kinase (ERK)-1/2 was performed. The role of the cell-ECM adhesion in the response of the chondroprogenitor cells to mechanical stress and the regulation of gene expression of focal adhesion kinase (FAK) and integrins were analyzed by using an in vitro system. A fibrous suture tissue replaced the midpalatal suture cartilage by the expansive force application for 14 days. The active osteoblasts that line the surface of bone matrix in the newly formed suture tissue strongly expressed the type I collagen gene, whereas they did not express the type II collagen gene. Although the numbers of precartilaginous cells expressing alpha5 and beta1 integrin increased, the immunoreactivity of alpha5 integrin in each cell was maintained at the same level throughout the experimental period. During the early response of midpalatal suture cartilage cells to expansive stimulation, formation of stress fibers, reorganization of focal adhesion contacts immunoreactive to a vinculin-specific antibody, and phosphorylation and nuclear translocation of ERK 1/2 were observed. In vitro experiments were in agreement with the results from the in vivo study, i.e. the inhibited expression of type II collagen and upregulation in integrin expression. The arginine-glycine-aspartic acid containing peptide completely rescued chondrogenesis from tension-mediated inhibition. Thus, we conclude that stretching activates gene expression of beta1 integrin and FAK and inhibits chondrogenesis through cell-ECM interactions of chondroprogenitor cells. PMID- 12751905 TI - Outer membrane protein A (OmpA) activates human epidermal Langerhans cells. AB - Outer membrane protein (Omp)A is highly represented and conserved in the Enterobacteriaceae family. Using a recombinant OmpA from Klebsiella pneumoniae (kpOmpA), we have analysed the interaction between this bacterial cell wall protein and human Langerhans cells (LC), the antigen-presenting cells of the epidermis and mucosa. We showed that biotinylated kpOmpA binds to human LC freshly isolated from epidermis. kpOmpA up-regulated MHC class II, CD86 and CCR7 expression, enhanced migration in response to macrophage inflammatory protein 3beta (MIP-3beta) through a reconstituted basement membrane mimicking the prerequisite passage through the dermal-epidermal basement membrane on the way to lymph nodes. The allostimulatory function of kpOmpA-treated LC was more potent than that of untreated cells. Even though the proportion of LC which binds kpOmpA was shown to vary between individuals, our data indicate that kpOmpA binds to and activates LC, and suggest that recognition of OmpA by LC may be an initiating event in the antibacterial host response. PMID- 12751906 TI - Multiple subphases of DNA repair and poly (ADP-ribose) synthesis in Chinese hamster ovary (CHO-K1) cells. AB - The two types of DNA synthesis as well as poly(ADP-ribose) biosynthesis were measured simultaneously in synchronized intact populations of CHO cells throughout the duration of S phase. Naturally occurring DNA fragmentation was detected by random primed oligonucleotide synthesis (ROPS assay). Fractions of synchronous cell populations were obtained by counterflow centrifugal elutriation. By gradually increasing the resolution of centrifugal elutriation multiple non-overlapping repair and replication peaks were obtained. The elutriation profile of DNA repair peaks corresponded to the DNA fragmentation pattern measured by ROPS assay. The number and position of poly(ADP-ribose) peaks during S phase resembled those seen in the DNA replication profile. Our results indicate that PAR synthesis is coupled to DNA replication serving the purpose of genomic stability. PMID- 12751907 TI - Effects of amifostine on liver oxidative stress caused by cyclophosphamide administration to rats. AB - Cyclophosphamide is an inactive cytostatic, which is metabolised into active metabolites mainly in the liver. During bioactivation, reactive oxygen species (ROS) are also formed, which can modify the components of both healthy and neoplastic cells leading to decreased antioxidative capacity. Amifostine is a drug that can inactivate ROS. The aim of the present study was to evaluate the influence of amifostine on the antioxidative system of the liver of rats exposed to cyclophosphamide. Intraperitoneal administration of cyclophosphamide was found to decrease the activity of liver antioxidative enzymes, i.e. superoxide dismutase, glutathione peroxidase and glutathione reductase, and to increase catalase activity. Amifostine slightly influenced antioxidative enzyme activity, causing a significant increase only in superoxide dismutase activity. Co administration of cyclophosphamide and amifostine nearly prevented changes in activities of superoxide dismutase, glutathione reductase and catalase, as well as to a high degree of glutathione peroxidase. Cyclophosphamide also evoked a decrease in the level of non-enzymatic antioxidants, such as reduced glutathione and vitamins C, E and A, as well as total antioxidant status. Administration of amifostine alone caused a significant increase in non-enzymatic antioxidant level that resulted in an increase in total antioxidant status. Administration of amifostine together with cyclophosphamide to a large extent prevented changes in the evaluated non-enzymatic antioxidative parameters, decreasing values of their concentration to the values of control group. Changes of liver antioxidative abilities during detoxification of cyclophosphamide were accompanied by intensified lipid peroxidation, manifested by an increase in concentration of products such as malondialdehyde and 4-hydroxynonenal. Amifostine caused the inhibition of lipid peroxidation in the liver of both control and cyclophosphamide-treated rats. In conclusion, our results suggest that amifostine significantly protects liver antioxidant properties from changes caused by cyclophosphamide treatment and in consequence prevents oxidative stress and phospholipid peroxidative damage. PMID- 12751908 TI - Effects of food on ethanol metabolism. AB - The goals of the present study were (1) to obtain ethanol pharmacokinetic data from fed dogs, and (2) perform Monte Carlo simulation to determine the effect of food on pharmacokinetic model parameter values. To a cohort of five fed dogs, 1 g ethanol per km body weight was administered as a gavage of 20% w/v ethanol solution. Blood samples taken at 0, 10, 20, 30, 40, 60, 80, 100, 120, 180, 240, and 360 minutes after the dose were mixed with anticoagulant and stored on ice. Blood ethanol concentration was determined via headspace chromatograph. Monte Carlo simulation with an ethanol pharmacokinetic model was used to estimate model parameter values and parameter standard deviations by minimization of the chi squared function. Results indicate that 50.6 +/- 21.0% of the ethanol dose was absorbed in the stomach, and an insignificant amount of ethanol was metabolized by gastric alcohol dehydrogenase postulated for the model. At 6 hours after the ethanol dose 59.4 +/- 21.0% of the ethanol dose was retained in the dogs' stomachs. PMID- 12751909 TI - Molecular modelling of the mouse cytochrome P450 CYP2F2 based on the CYP102 crystal structure template and selective CYP2F2 substrate interactions. AB - The results of homology modelling of the mouse cytochrome P450, CYP2F2, are reported, based on the CYP102 crystallographic template. It is found that selective CYP2F2 substrates are able to fit the putative active site of the enzyme via aromatic pi-pi stacking and, in some cases, hydrogen-bonded interactions. Two alkylnaphthalenes were investigated via the model to evaluate whether they are likely to act as CYP2F2 substrates and, of these, 2-isopropyl naphthalene was found to fit the putative active site, whereas 2-(2 hexadecyl)naphthalene was unable to do this, due to its bulky side-chain. Consequently, it is possible to utilize homology modelling to evaluate the likelihood of enzyme-substrate selectivity for CYP2F2 and predict routes of metabolism mediated by this enzyme. This procedure can therefore be used to investigate the potential for activation of xenobiotics via this enzyme, especially those related to known CYP2F substrates, such as naphthalene. PMID- 12751910 TI - Molecular modelling of CYP2B6 based on homology with the CYP2C5 crystal structure: analysis of enzyme-substrate interactions. AB - The results of homology modelling of CYP2B6 based on the CYP2C5 crystal structure is described in terms of substrates and inhibitors binding within the putative active site. In general these results are in agreement with currently available evidence from substrate metabolism, mode of inhibitor action and site-directed mutagenesis experiments within the CYP2B subfamily of enzymes. Consequently, the model based on the CYP2C5 template represents an advance on those models produced from bacterial P450s, such as CYP101 and CYP102. Quantitative Structure-Activity Relationships (QSARs) for substrates binding to CYP2B6 indicate a key role for hydrogen bonding, and lipophilic character, as determined by the log P parameter (where P is the octanol/water partition coefficient), is also of importance for explaining the variation in experimental binding affinity for CYP2B6 substrates. It is possible to estimate the binding energies for typical CYP2B6 substrates based on their properties and interactions with the enzyme, which show good concordance with experimental data in the form of apparent Km values. PMID- 12751911 TI - Influence of multiple dose activated charcoal on the disposition kinetics of irinotecan in rats. AB - INTRODUCTION: The main clinical adverse effect of irinotecan (CPT-11) therapy is diarrhoea. Using a rat model, we attempted to study the effects of activated charcoal on the prevention of diarrhoea after a bolus dose of CPT-11, and also investigated the disposition kinetics of CPT-11 and its metabolite, SN-38, as well as SN-38 glucuronide (SN-38G) in the presence and absence of charcoal. MATERIALS AND METHODS: Male Sprague-Dawley rats were given a daily oral dose of activated charcoal (Ultracarbon, 2.5 g/kg daily for 5 days) 10 min before an i.v. bolus injection of CPT-11 (60 mg/kg daily for 5 days; total dose 300 mg/kg); the control group was given CPT-11 alone. The pharmacokinetics of CPT-11, SN-38 and SN-38G were determined in both groups of rats on day 1. The incidence of diarrhoea was monitored throughout the course of the study. RESULTS: There were no differences in the mean (+/- SD) C, (15.8 +/- 7.5 vs 12.1 +/- 3.3 microg/ml), t1/2 (2.2 +/- 0.7 vs 2.2 +/- 0.5 h), CL (5.7 +/- 2.1 vs 6.8 +/- 1.2 l/h/kg), Vd (1798 +/- 958 vs 2280 +/- 731 l/kg) or AUC0-infinity (11.8 +/- 3.9 vs 9.1 +/- 1.7 microg x h/ml) of CPT-11 after dosing with or without activated charcoal. Similarly, charcoal treatment had no effect on the disposition kinetics of SN-38 and SN-38G. A higher frequency of grade 3 diarrhoea was observed in the control group compared to the charcoal treatment group (log OR: -1.06; 95% CI: -2.25, 0.13) but this was only marginally statistically significant (p = 0.08). CONCLUSION: These results suggest that multiple oral doses of activated charcoal do not modulate the clearance of CPT-11 and SN-38 in rats. The implication is that activated charcoal alone may not be very effective in preventing CPT-11 induced diarrhoea. PMID- 12751912 TI - Quality of life and economic considerations in the management of prostate cancer. AB - The purpose of this article was to provide an overview of the morbidity and mortality of prostate cancer, QOL issues and the economic impact of the disease. We searched Medline (from 1990 onwards) for all studies dealing with prostate cancer epidemiology, treatment, screening and staging, and critically reviewed the most relevant articles, focusing on pharmacoeconomic issues. Prostate cancer is the most common cancer in men. In the US, new estimated cases of prostate cancer represented 14.8% of all new cancer cases for 2000, with estimated deaths from prostate cancer comprising 5.8% of all deaths from cancer. Current options for prostate cancer management include radical prostatectomy, cryosurgery, radiotherapy, hormone therapy and watchful waiting. Many of the long-term effects of treatment, such as urinary incontinence, impotence and radiation-induced proctitis, have a large impact on patients' quality of life and, in some patients, may offset the clinical benefits. Regulatory bodies and managed care organisations are assigning increasing importance to the evaluation of QOL benefits as an independent clinical endpoint and a measure of patient satisfaction. Several screening programmes for early detection of prostate cancer, mostly based on prostate-specific antigen (PSA) measurement or digital rectal examination, have been proposed, but their routine implementation in all asymptomatic elderly men has been questioned. There is still no definite proof that patient outcomes are improved by extensive PSA screening. Furthermore, the total cost of a screening programme is difficult to define since it extends well beyond the initial test. Several instruments are used for QOL assessment in prostate cancer, some of which have been specifically developed for, or adapted to, patients with this disease, such as the Functional Assessment Cancer Therapy (FACT) tool, Prostate Cancer Treatment Outcome Questionnaire (PCTO-Q) and Prostate Cancer Specific Quality of Life Instrument (PROSQOLI). More than 50% of treatment costs for prostate cancer are accrued during the patient's last year of life, and total initial care costs decrease with increasing age. In the US, initial average inpatient costs were estimated at $US 2253, in 1995, for men aged > or =80 years, compared with $US 4540 for men aged 35-64 years. In recent years, treatments based on combined modalities (i.e. radiotherapy/prostatectomy plus hormonal therapies) have emerged. Although cost-effectiveness analyses of various treatment options have been attempted, the strength of their conclusions appears to be limited by the lack of homogeneous literature data on the effects of such interventions on survival and morbidity. PMID- 12751913 TI - The economics of screening and treatment in type 2 diabetes mellitus. AB - A systematic review of the literature was conducted to identify articles on the economics of type 2 diabetes mellitus. Articles were classified into two main categories: cost/burden-of-illness studies of type 2 diabetes and economic evaluations of type 2 diabetes interventions. This systematic review was supplemented by an overview of the findings relating to economic evaluations of associated diabetic complications. A number of conclusions emerge from this review, the most important of which is that intensive treatment of patients with type 2 diabetes appears to be relatively cost effective compared with more conservative strategies. This finding reflects the cost offsets that arise from the range and degree of complications attributable to diabetes. Primary prevention of type 2 diabetes also appears to be cost effective, particularly in high-risk groups. The evidence on screening for type 2 diabetes is less conclusive and further economic analysis is required. PMID- 12751914 TI - Labour force participation among individuals with hepatitis C in the US. AB - BACKGROUND: In 1996, the number of persons newly infected with hepatitis C virus (HCV) in the US was estimated to be 36 000. As a chronic disease that primarily affects younger persons, hepatitis C has the potential to influence employment considerably. OBJECTIVE: To estimate employment effects associated with hepatitis C morbidity. DESIGN: An economic model of labour supply, which used the outcome measure workforce participation (yes/no), was applied. STUDY PARTICIPANTS: The study samples (by gender) were comprised of persons 18-65 years of age, with and without serological evidence of HCV infection, and with normal or elevated levels of alanine aminotransferase (ALT) who participated in the National Health and Nutrition Examination Survey III from 1988-1994. RESULTS: After controlling for the potential confounding effects of demographic, social, and economic factors, positive HCV status/normal ALT level in males was associated with a 10.7% reduction in labour force participation (when compared with negative HCV status). Positive HCV status and elevated ALT levels was associated with a 17.5% reduction in employment. The results for females were not statistically significant. CONCLUSIONS: Nationally, the employment response for HCV-positive status and elevated ALT levels translates into an excess non-employment of 48 000 males annually. PMID- 12751915 TI - The cost effectiveness of ACE inhibitors as first-line antihypertensive therapy. AB - BACKGROUND: Current hypertension guidelines differ in their recommendations for first-line antihypertensive therapy. OBJECTIVE: To evaluate the cost effectiveness of ACE inhibitor therapy as antihypertensive first-line therapy as compared with conventional antihypertensive therapy with beta-adrenoceptor antagonists or diuretics. STUDY DESIGN: Cost-effectiveness analysis based on data from randomised trials and observational studies comparing the effectiveness of ACE inhibitor and conventional antihypertensive therapy, we constructed a Markov model to compare four strategies in the management of uncomplicated hypertension: (i) prescribing ACE inhibitor therapy to all patients; (ii) prescribing conventional therapy to all patients; (iii) individualised antihypertensive therapy based on the presence or absence of left ventricular hypertrophy on electrocardiography (ECG); or (iv) individualised antihypertensive therapy based on the presence or absence of left ventricular hypertrophy on echocardiography. METHODS: Cost data were derived from the medical literature and focus groups, and utility values were derived from patients on antihypertensive monotherapy. All costs were calculated in 1999 Canadian dollars, but are reported in US dollars according to the 1999 purchasing power parity rate for medical and healthcare. The effectiveness of ACE inhibitor therapy in the presence of left ventricular hypertrophy was derived from observational studies. The time horizon was over a lifetime. PERSPECTIVE: Third-party payer. PATIENTS/PARTICIPANTS: A cohort of men aged 40 years without cardiovascular comorbidity requiring antihypertensive drug therapy. MAIN OUTCOME MEASURES AND RESULTS: In the baseline analysis, all four strategies resulted in expected discounted QALYs that differed from each other only at the third decimal point (i.e. less than 0.003). Given the uncertainties in the variable estimates and the small size of the differences, these differences are extremely small and unlikely to represent real differences. Even accepting the small gains as real, the resulting cost-effectiveness ratios are unattractively high: $US 200,000 per QALY gained for the echocardiography strategy (compared with ECG), and $US 700,000 for the "ACE inhibitor for all" strategy (compared with ECG). The incremental cost effectiveness of prescribing ACE inhibitor therapy to everybody was never less than $US 100,000/QALY in the sensitivity analysis. CONCLUSIONS: Prescribing ACE inhibitors as antihypertensive first-line therapy in patients without cardiovascular morbidity cannot be recommended at the present time unless the acquisition costs of ACE inhibitors become substantially more attractive. PMID- 12751916 TI - Cost effectiveness of pneumococcal vaccination for infants and children with the conjugate vaccine PnC-7 in Germany. AB - BACKGROUND: The introduction of the conjugate vaccine PnC-7 implies that a pneumococcal vaccine is available, for the first time, which also gives children under the age of 2 years reliable protection against invasive pneumococcal infections and offers some protection against non-invasive pneumococcal infections. OBJECTIVE AND PERSPECTIVE: In the context of a multiple-period Markov model, a cost-effectiveness analysis of a recommendation for general pneumococcal vaccination in Germany for infants and children under the age of 2 years was performed from the healthcare payer, public authority and societal perspectives. DESIGN: Various published data on age-specific incidence rates, mortality rates, efficacy of the conjugate vaccine PnC-7 and treatment costs of pneumococcal infections were incorporated into a Markov model to quantify the consequences of vaccinating versus not vaccinating. RESULTS: From a German healthcare payers' perspective, general vaccination with the conjugate vaccine would redeem 51.1% of the vaccination costs due to avoidable treatment costs, whereas, from a broader point of view, the benefits, expressed in monetary terms, would exceed the cost of vaccination. The conjugate vaccine would require an investment of euro72 866 per life-year saved (discounted, healthcare payers' viewpoint). Besides this benchmark, there are further outcome measures which cannot be ignored by those deciding on a general vaccination recommendation: 450 000 preventable episodes of illness and 134 cases of sequelae which can be prevented. CONCLUSIONS: The vaccination with the conjugate vaccine PnC-7 is cost saving from a broader perspective and the results should not be ignored by policy makers in regard to a general vaccination recommendation. PMID- 12751917 TI - Vaccines for Alzheimer's disease: how close are we? AB - Alzheimer's disease is a neurodegenerative disorder characterised by a progressive loss of cognitive function. Despite the considerable progress being made, a complete description of the molecular pathology of this disease has yet to be elucidated. The evidence indicates that abnormal processing and extracellular deposition of the longer form of the beta-amyloid (Abeta) peptide (Abeta(1-42), a proteolytic derivative of the amyloid precursor protein [APP]) is implicated in the pathogenesis of Alzheimer's disease. In this respect, recent use of experimental mouse models, in which the mice develop some aspects of Alzheimer's disease in a reproducible fashion, has provided a new opportunity for a multidisciplinary and invasive analysis of mechanisms behind the amyloid pathology and its role in Alzheimer's disease. It has been demonstrated, using a single transgenic mouse model system that overexpresses the human mutated APP gene, that an immunisation against Abeta(1-42) causes a marked reduction in the amyloid burden in the brain. The follow-up research provided more evidence that both active and passive Abeta immunisation also reduces cognitive dysfunction in transgenic mouse models of Alzheimer's disease. Other studies using different approaches - such as secretase, cholesterol and Abeta metalloprotein inhibitors or NSAIDs - but all targeting the abnormal metabolism of Abeta have confirmed in each case that a significant reduction of amyloid plaque burden can be achieved in transgenic mouse models of Alzheimer's disease. This research strongly supports the notion that abnormal Abeta processing is essential to the pathogenesis of Alzheimer's disease and provides a crucial platform for the development and detailed testing of potential treatments in experimental models before each of these approaches can be proposed as a therapy for Alzheimer's disease. Although the first clinical trial of active immunisation with a pre aggregated synthetic Abeta(42) preparation (AN-1792 vaccine) met with some setbacks and was discontinued after several patients experienced meningoencephalitis, the follow-up analysis of the effect of immunisation against Abeta in humans revealed a powerful effect of vaccination in the clearance of amyloid plaques from the cerebral cortex. PMID- 12751918 TI - Levodopa-induced response fluctuations in patients with Parkinson's disease: strategies for management. AB - Fluctuations in response to levodopa in patients in the advanced stages of idiopathic Parkinson's disease occur frequently and are a difficult problem to treat. Patients who are treated with levodopa have an additional 10% risk of experiencing response fluctuations with each year of treatment: 50% of patients have this problem after 5 years of receiving levodopa therapy and almost 100% of patients after 10 years. The mechanisms by which response fluctuations occur are only partially understood and can be divided into three main types: (i) presynaptic neuronal degeneration leading to a lack of buffering of released levodopa, which is mainly related to wearing-off phenomena; (ii) postsynaptic changes in dopamine receptor sensitivity and number, partially caused by the presynaptic changes, which are clinically related to at-random response fluctuations; and (iii) pharmacokinetic and pharmacodynamic influences of exogenously administered dopaminergic agents. Several oral and parenteral treatment strategies are recommended to manage response fluctuations, such as optimisation of dopamine receptor agonist therapy in combination with a reduction of the levodopa load; use of slow-release levodopa formulations; use of catechol O-methyltransferase inhibitors; an increase of levodopa dose frequency; use of high-dose amantadine; and intermittent or continuous use of apomorphine and/or levodopa. Continuous stimulation of dopamine receptors with dopaminergic agents is one of the crucial basic steps in the treatment of patients at an advanced stage of Parkinson's disease, and the preferential use of dopamine receptor agonists has proven to be successful in the prevention and treatment of response fluctuations. PMID- 12751922 TI - Early and late functional outcome assessments following endovascular and open aneurysm repair. AB - PURPOSE: To compare early and late functional outcomes, as well as survival and recovery, following endovascular or open repair of abdominal aortic aneurysm (AAA). METHODS: Between 1996 and 2000, 294 patients underwent AAA repair (141 open and 153 endovascular); 57 patients from each group had 12-month follow-up for functional outcome assessment. Recovery was measured as hospital length of stay, skilled nursing requirement, and hospital readmission within 1 year to determine cumulative hospital utilization. Early (<6 months) functional outcomes were measured by activity level and convalescence days following surgery. Late (>6 months) functional outcomes were measured as ambulation, independent living, and employment status pre- and postoperatively. RESULTS: Operative mortality for open repair was 5 (3.5%) compared to 1 (0.6%) after an endovascular procedure (p<0.05). The endovascular group had a shorter hospital stay (2.8+/-2.8 versus 8.3+/-4.5 days) and fewer skilled nursing requirements (0% versus 26%; p<0.001). Cumulative hospital utilization over 12 months was 3.8 days for endovascular patients and 13.8 days for open repair (p<0.001). Recovery time was 99.3+/-84.1 days (range 14-365) in conventionally treated patients and 32.1+/-43.5 days (range 7-180) in the stent-graft group (p<0.001). At 6 months, 43 (75%) open and 54 (95%) endovascular patients had full recovery (p<0.01). Activity levels decreased in 13 (23%) open and 3 (5%) endovascular patients after surgery (p<0.01). There were no differences in ambulation, independent living, or employment status before and after treatment. CONCLUSIONS: Periprocedural survival following aneurysm repair is improved with endovascular grafting compared to open surgery, and recovery is more rapid, with a 78% reduction in total hospital days. Early functional outcomes are markedly improved with endovascular repair, while there is no difference in late functional outcomes between the procedures. PMID- 12751923 TI - Delayed complications after endovascular AAA repair in women. AB - PURPOSE: To assess whether women suffer more delayed complications or require more interventions after endovascular abdominal aortic aneurysm (AAA) repair. METHODS: Over a 2-year period, 245 patients (203 men, 42 women) underwent endovascular repair with the AneuRx stent-graft at our institution. Baseline, operative, and follow-up data were reviewed and outcomes compared between men and women for complications or secondary procedures beyond the 30-day postoperative period. RESULTS: The 42 (17.1%) women treated with the AneuRx graft experienced more technical complications (17% versus 8.3%; p<0.05) and acute conversions to open repair (6/7, 86%; p=0.0005). At 30 days, systemic morbidity was significantly higher in women (26% versus 5.3%; p=0.001.) However, mortality (2.3% versus 1.5%), length of hospital stay (2.6 versus 2.9 days), and major endoleaks at discharge (0% versus 1.6%) were similar. Over a mean follow-up of 11.3 months (range 1-26), graft limb occlusion occurred more frequently in female patients (12.2% versus 2.5%; p=0.05), and secondary procedures were required in a significantly higher number of women (29% versus 9%; p<0.05). Three patients in each group (25% of women versus 17% of men) who had secondary procedures required additional interventions. Although not statistically significant, the mortality in women during follow-up was higher (9.7% versus 5%); 2 deaths were directly related to graft complications in the female group. CONCLUSIONS: Endoluminal AAA repair in women presents a technical challenge at the time of operation and may cause more frequent late complications, necessitating close postprocedural surveillance and prompt correction of complications to avoid potentially catastrophic outcomes. Reassessment of inclusion criteria for women may be necessary. PMID- 12751924 TI - Endovascular abdominal aortic aneurysm repair is less invasive, now we must prove its efficacy. PMID- 12751919 TI - Suicidal behaviour in bipolar disorder: risk and prevention. AB - Bipolar (manic-depressive) disorder is a common and severe illness. It is also potentially fatal as a result of accidents and increased mortality associated with comorbid substance use and medical illnesses, but its highest lethality results from suicide. Suicide rates, averaging 0.4% per year in men and women diagnosed with bipolar disorder, are >20-fold higher than in the general population. Suicidal acts often occur early in the illness course and in association with severe depressive and dysphoric-agitated mixed phases of illness, especially following repeated, severe depressions. Systematic consideration of risk and protective factors enhances assessment of potentially suicidal patients. Short-term interventions employed empirically to manage acute suicidality include close clinical supervision, rapid hospitalisation and use of electroconvulsive treatment. Several plausible therapeutic interventions have limited evidence of long-term effectiveness against mortality risks associated with any psychiatric disorder, including antidepressant, antimanic, antipsychotic and electroconvulsive, as well as psychosocial, treatments. However, in bipolar disorder and other major affective disorders, lithium maintenance treatment is a notable exception, with strong and consistent evidence that it reduces suicidal risk. The growing range of drugs being introduced to treat acute and long-term phases of bipolar disorder, including antiepileptic drugs, atypical antipsychotics and relatively safe, modern antidepressants, require research assessment for their ability to limit premature mortality from suicide and other causes. For now, however, more can be done to improve treatment in major affective illnesses by application of current knowledge in a systematic fashion, with close and sustained clinical follow-up of patients at risk, hopefully with a resulting reduction of mortality rates. PMID- 12751925 TI - Clinical application of a curved nitinol stent-graft for thoracic aortic aneurysms. AB - PURPOSE: To evaluate the clinical efficacy of a curved nitinol stent-graft for repair of thoracic aortic aneurysms. METHODS: The Matsui-Kitamura stent-graft (MKSG), composed of a self-expanding nitinol stent and polyester fabric, was shaped to match the aortic curvature of 11 patients (6 men; mean age 72.6 years, range 33-90) with 6 true and 5 false aneurysms of the distal arch or proximal descending aorta. The delivery system was an 18 or 20-F J-shaped sheath combined with a preloader-type introducer. The original mean proximal neck length was 16.4 mm, but 4 patients received an axilloaxillary bypass to lengthen the neck. Although the mean corrected proximal neck length was 21.9 mm (overall), 5 cases still had proximal necks <15 mm long. RESULTS: All curved MKSGs were successfully deployed in the correct position and fitted to the curvature of the aortic arch, achieving complete aneurysm exclusion in 8 (73%) cases. The other 3 repairs displayed early endoleaks; 1 received an additional MKSG, but the other 2 are being observed. Thirty-day mortality was 0%. One patient developed transient renal failure requiring hemodialysis; no neurological complications were observed. CONCLUSIONS: Endovascular repair of thoracic aortic aneurysms using curved MKSGs appears to be feasible and clinically effective. A tighter fit of the device to the curvature of the aortic arch may exclude distal arch aneurysms despite a short proximal neck. PMID- 12751926 TI - Successful endoluminal repair of an infected thoracic pseudoaneurysm caused by methicillin-resistant Staphylococcus aureus. AB - PURPOSE: To report the successful endoluminal repair of an infected thoracic aneurysm secondary to methicillin-resistant Staphylococcus aureus (MRSA). CASE REPORT: A 76-year-old man presented with an infected thoracic pseudoaneurysm 9 weeks after an elective infrarenal aneurysm repair. Blood cultures were positive for MRSA. Computed tomography (CT) showed an 11.5-cm false aneurysm of the descending thoracic aorta just proximal to the celiac axis. An Excluder stent graft was used to successfully repair the lesion. Recovery was uneventful, and the patient was treated with linezolid for 6 weeks. Follow-up CT scans at 3 and 12 months confirmed exclusion of the aneurysm and progressive shrinkage of the aneurysm sac with no evidence of graft infection. CONCLUSIONS: Endoluminal repair is an alternative to open surgery for the treatment of infected aneurysms of the thoracic aorta. PMID- 12751920 TI - Clinically important drug interactions with zopiclone, zolpidem and zaleplon. AB - Insomnia, an inability to initiate or maintain sleep, affects approximately one third of the American population. Conventional benzodiazepines, such as triazolam and midazolam, were the treatment of choice for short-term insomnia for many years but are associated with adverse effects such as rebound insomnia, withdrawal and dependency. The newer hypnosedatives include zolpidem, zaleplon and zopiclone. These agents may be preferred over conventional benzodiazepines to treat short-term insomnia because they may be less likely to cause significant rebound insomnia or tolerance and are as efficacious as the conventional benzodiazepines. This review aims to summarise the published clinical drug interaction studies involving zolpidem, zaleplon and zopiclone. The pharmacokinetic and pharmacodynamic interactions that may be clinically important are highlighted. Clinical trials have studied potential interactions of zaleplon, zolpidem and zopiclone with the following types of drugs: cytochrome P450 (CYP) inducers (rifampicin), CYP inhibitors (azoles, ritonavir and erythromycin), histamine H(2) receptor antagonists (cimetidine and ranitidine), antidepressants, antipsychotics, antagonists of benzodiazepines and drugs causing sedation. Rifampicin significantly induced the metabolism of the newer hypnosedatives and decreased their sedative effects, indicating that a dose increase of these agents may be necessary when they are administered with rifampicin. Ketoconazole, erythromycin and cimetidine inhibited the metabolism of the newer hypnosedatives and enhanced their sedative effects, suggesting that a dose reduction may be required. Addition of ethanol to treatment with the newer hypnosedatives resulted in additive sedative effects without altering the pharmacokinetic parameters of the drugs. Compared with some of the conventional benzodiazepines, fewer clinically important interactions appear to have been reported in the literature with zaleplon, zolpidem and zopiclone. The fact that these drugs are newer to the market and have not been as extensively studied as the conventional benzodiazepines may be the reason for this. Another explanation may be a difference in CYP metabolism. While triazolam and midazolam are biotransformed almost entirely via CYP3A4, the newer hypnosedatives are biotransformed by several CYP isozymes in addition to CYP3A4, resulting in CYP3A4 inhibitors and inducers having a lesser effect on their biotransformation. PMID- 12751927 TI - Evaluation of glycoprotein IIb/IIIa inhibitors in carotid angioplasty and stenting. AB - PURPOSE: To review the immediate neurological and bleeding complications associated with the use of glycoprotein (GP) IIb/IIIa inhibitors in patients undergoing extracranial carotid artery stent placement. METHODS: A retrospective review was performed of 550 patients (321 men; mean age 71.1 years, range 28-91) who underwent carotid artery angioplasty and stent placement. Glycoprotein IIb/IIIa inhibitors were given prophylactically along with heparin to 216 patients, whose outcomes were compared to a control group of 334 patients who received intravenous heparin alone. Primary endpoints were the immediate and 30 day neurological complications, including transient ischemic attacks (TIAs), minor and major strokes, and neurologically-related deaths. The secondary endpoint was any abnormal bleeding. RESULTS: The all stroke/neurological death rate in 216 patients treated with heparin and GP IIb/IIIa inhibitors was 6.0% (13 events) compared 2.4% (8 events) in the 334 patients in the heparin-only control group (p=0.0430). Two of the 4 neurologically-related deaths in the GP IIb/IIIa inhibitor group resulted from intracranial hemorrhages; there were no intracranial hemorrhages in the heparin-only group. There was 1 episode of extracranial bleeding in the GP IIb/IIIa inhibitor group treated with embolization. The incidences of significant puncture-site bleeding requiring transfusion were similar in the groups. CONCLUSIONS: Neurological complications following percutaneous carotid artery interventions have been relatively few. The neurological sequelae in carotid stent patients receiving glycoprotein IIb/IIIa inhibitors were more numerous and consequential, which suggests that the use of GP IIb/IIIa inhibitors in carotid stenting should be discouraged. PMID- 12751928 TI - Adjunctive use of platelet glycoprotein IIb/IIIa inhibitors for carotid angioplasty and stent placement: time to say good bye? PMID- 12751929 TI - Endovascular management of acute carotid artery dissection with a waxing and waning neurological deficit. AB - PURPOSE: To evaluate the feasibility and efficacy of emergent carotid stenting for an acute internal carotid artery (ICA) dissection. CASE REPORT: A 51-year-old man was admitted to our emergency department's stroke unit 1 hour after the onset of left hemiparesis. Computed tomographic and transcranial Doppler scans showed no pathological findings, but the color Doppler study detected a double lumen in the right carotid bifurcation extending to the proximal ICA. Within the first 2 hours after admission, the patient's neurological status began to fluctuate; the NIH Stroke Scale (NIHSS) and Rankin scores evaluated each hour after admission ranged from 0 to 12 and from 0 to 3, respectively. Emergency carotid angiography confirmed the dissection of the bulb and proximal right ICA, which prompted implantation of 2 Wallstents from the internal to common carotid artery. Forty eight hours later, the patient was almost totally asymptomatic with an NIHSS score of 1 and a Rankin score of 0; he was discharged on postoperative day 3. At 3 months, the patient was free of neurological symptoms and the stented carotid artery was patent. CONCLUSIONS: In patients with fluctuating neurological signs and symptoms consistent with carotid artery dissection that are refractory to medical therapy, a stent can be placed to prevent permanent neurological deficits provided that the anatomical conditions are suitable. PMID- 12751930 TI - Recurrent vertebral arteriovenous fistula after surgical repair: treatment with a self-expanding stent-graft. AB - PURPOSE: To describe successful endovascular repair of a recurrent vertebral arteriovenous fistula (AVF) after surgical correction. CASE REPORT: A 42-year-old woman presented with recurrent dizziness, nausea, and headaches. A loud bruit in the right neck was present. Central venous catheter insertion had been done 1 year previously, creating an angiographically documented right-sided vertebral AVF that was successfully excluded by a surgical procedure. Four weeks later, the AVF reappeared. Successful endovascular repair with a self-expanding stent-graft was performed. Follow-up over 12 months was uneventful, with a patent vertebral artery and no recurrence of symptoms. CONCLUSIONS: Endovascular stent-graft repair is feasible and offers a therapeutic alternative in the treatment of vertebral AVF, in particular for recurrence after initial surgery. This minimally invasive method may become the treatment of choice in the management of such lesions, preserving patency of the vertebral artery. PMID- 12751931 TI - Endovascular management of impending carotid rupture in a patient with advanced head and neck cancer. AB - PURPOSE: To describe an innovative endovascular technique combining a flexible self-expanding stent-graft to protect the common and internal carotid artery with selective coil embolization of the affected external carotid artery (ECA) branches. CASE REPORT: A 42-year-old man presented with episodes of profuse but self-limited carotid hemorrhage from advanced squamous cell carcinoma of the neck. Despite invasion of the carotid sheath by the tumor, angiography confirmed the source of bleeding to be invasion of branches of the ECA. Successful coil embolization of the ECA and stent-graft isolation of the common/internal carotid arteries were performed. At 1-month follow-up, carotid angiography confirmed the complete occlusion of the ECA and patent stent-graft. There was no recurrent bleeding or neurological deficits at 6 months after the procedure. CONCLUSIONS: Stent-grafts in combination with coil embolization can be used to manage patients with impending carotid rupture secondary to head and neck cancer. PMID- 12751932 TI - Elective and emergent endovascular treatment of subclavian artery aneurysms and injuries. AB - PURPOSE: To demonstrate our short and long-term results after transbrachial treatment of subclavian artery aneurysms and injuries with stent-grafts in elective and emergency settings. METHODS: Ten of 12 consecutive patients (6 men; mean age 63.8 years, range 38-80) were treated electively with commercially prepared endografts delivered via a transbrachial access to repair a subclavian artery aneurysm (n=3) or an injury from a misplaced central venous catheter (n=7). Two patients required emergency treatment for a ruptured atherosclerotic aneurysm in one and an unintentional arterial puncture during placement of a central venous access in the other. Stent-graft patency during follow-up was assessed by physical examination with comparison of brachial blood pressures in all patients; computed tomography angiography (CTA) was performed in available patients. RESULTS: Successful deployment of stent-grafts with sealing of the lesion was achieved in all cases. There were 2 (17%) procedural complications. One patient developed an access-site hematoma that required surgical revision. The second patient, who had a right subclavian injury, suffered an embolic cerebral infarction. The primary stent-graft patency during follow-up (mean 11.6 months) was 100%. CTA examinations in 7 patients at a mean 18 months showed strut dislocation at the thoracic outlet without luminal narrowing in 1 patient. A 50% intraluminal narrowing due to compression between the clavicle and the first rib occurred in another patient. Six patients with a mean follow-up of 23 months (range 0.3-4.5 years) are still alive with patent stent-grafts. CONCLUSIONS: Endovascular stent-graft treatment of subclavian artery aneurysms and injuries is a less invasive alternative to surgical repair. Long-term results must still be confirmed in further studies. PMID- 12751933 TI - Stent-graft repair of a mycotic left subclavian artery pseudoaneurysm. AB - PURPOSE: To report successful stent-graft treatment of a mycotic pseudoaneurysm of the left subclavian artery in an immunosuppressed patient. CASE REPORT: A 17 year-old immunosuppressed woman undergoing treatment for recurrent leukemia developed persistent fever and 2 episodes of hemoptysis. A contrast-enhanced computed tomographic (CT) scan demonstrated a saccular aneurysm of the left subclavian artery, which was considered to be a mycotic aneurysm caused by erosive fungal infection from the lung. The pseudoaneurysm was treated with a homemade stent-graft consisting of a nitinol stent and a polyester fabric. A type II endoleak present at the end of the procedure appeared to have sealed spontaneously on the CT scan at 3 days. No neurological deficit or ischemic symptoms of the left arm were noted during the follow-up, which lasted until the patient died 11 months later after rejecting a second bone marrow transplant. CONCLUSIONS: Endovascular repair may be an alternative to open surgery for the management of mycotic aneurysms of the subclavian artery. PMID- 12751934 TI - Endovascular repair of a ruptured renal artery aneurysm. AB - PURPOSE: To describe the percutaneous treatment of a ruptured renal artery aneurysm (RAA) using a stent-graft. CASE REPORT: An 86-year-old woman had sudden onset of right-sided back pain and a swollen left leg. Computed tomography disclosed a right retroperitoneal mass involving the kidney; the inferior vena cava was compressed, with thrombus in the left common iliac artery. There was a suggestion of RAA associated with contrast extravasation. Angiography confirmed a saccular aneurysm of the distal right main renal artery immediately proximal to the first segmental branch. A Jostent peripheral stent-graft was implanted with complete exclusion of the lesion. At 6-month follow-up, the patient remained asymptomatic, but there was a suggestion of in-stent restenosis, which is being followed closely. CONCLUSIONS: In elderly patients who are hemodynamically stable and have aneurysm anatomy that is suitable for stent-graft placement, the endovascular approach may allow rapid and definitive treatment of the hemorrhage. PMID- 12751935 TI - Endovascular therapy combined with immunosuppressive treatment for pseudoaneurysms in patients with Behcet's disease. AB - PURPOSE: To evaluate the feasibility, efficacy, and outcome of endovascular therapy combined with immunosuppression for the treatment of arterial pseudoaneurysms due to Behcet's disease. METHODS: Eleven pseudoaneurysms (3 in the abdominal aorta, 3 in the subclavian artery, and individual lesions in the brachiocephalic artery, renal artery, common iliac artery, common carotid artery, and the descending thoracic aorta) in 9 patients with Behcet's disease were treated with 10 stent-grafts and 1 self-expanding stent. All patients with elevated erythrocyte sedimentation rate (ESR) were treated with immunosuppressive agents (azathioprine, prednisolone) before and after the procedure. RESULTS: Endovascular treatment was successful in all cases, without major adverse events. The ESR was reduced from 42.7+/-18.0 mm/h initially to 18.6+/-12.6 mm/h after immunosuppressive therapy prior to endovascular repair. During follow-up (mean 24.1+/-14.0 months, range 6-43), 8 of 11 lesions showed complete resolution. One stent-graft to treat a postsurgical recurrent carotid artery pseudoaneurysm was occluded, and an abdominal aortic pseudoaneurysm recurred; both patients stopped their medications. ESR during follow-up was 7.9+/-4.5 mm/h. CONCLUSIONS: Endovascular treatment for pseudoaneurysms due to Behcet's disease is feasible and effective when disease activity is strictly controlled with immunosuppressive therapy. PMID- 12751936 TI - Thrombolysis of occluded synthetic bypass grafts in the lower limb: technical success and 1-year follow-up in 32 patients. AB - PURPOSE: To evaluate prospectively the technical success and clinical outcome of thrombolysis for acute occlusion of synthetic arterial bypass grafts in the lower limb. METHODS: Thirty-two consecutive patients (27 men; median age 65 years, range 41-80) with occluded polytetrafluoroethylene bypass grafts were treated with direct-catheter thrombolysis (100,000-IU bolus of urokinase with 100,000 IU/h infusion) followed by ancillary interventions to treat underlying stenosis whenever necessary. All patients received oral anticoagulation to maintain the international normalized ratio at 3.0 to 4.0. Clinical follow-up and duplex ultrasound examinations were performed at 3-month intervals up to 1 year. RESULTS: Thrombolysis was technically successful in 27 (84%) patients; 3 of the 5 failed patients had amputations. Mean duration of urokinase therapy was 36+/-14 hours. In 18 patients, underlying stenoses (11 distal anastomosis, 5 proximal anastomosis, and 3 inflow) were treated, 15 by an endovascular procedure and 3 surgically. Four major complications occurred: groin hematoma, sepsis, transient renal dysfunction, and a hemorrhage at the proximal anastomosis after urokinase treatment. At 1 year, 21 bypass grafts had reoccluded (20% patency rate on intention-to-treat basis); 3 reocclusions resulted in amputation (overall 19% amputation rate). CONCLUSIONS: Thrombolysis in the setting of acute lower limb bypass graft occlusion is associated with good initial technical success rates and satisfactory clinical results. However, the re-occlusion rate within 1 year is high. PMID- 12751937 TI - Regional thrombolytic therapy and stent implantation in an acutely occluded aortobifemoral bypass graft. AB - PURPOSE: To present a case highlighting the efficacy of local thrombolysis followed by stent implantation in acute occlusion of an aortobifemoral bypass graft. CASE REPORT: A 47-year-old man was referred to our catheterization laboratory for acute bilateral limb ischemia occurring 18 hours after an aortobifemoral bypass procedure. Angiography documented total occlusion due to massive thrombosis of the grafts. After 24 hours of local thrombolysis, repeat angiography showed complete lysis of the thrombus and an intimal flap in the abdominal aorta. A Palmaz stent was successfully implanted to cover the flap and restore adequate flow. At 1 year, the patient remains asymptomatic, and angiography showed patency of both the stent and the grafts. CONCLUSIONS: Based on this patient's response, local thrombolysis followed by stenting may be a safe and effective alternative to reoperation for the treatment of acute graft thrombosis caused by an intimal aortic flap. More experience with this approach is necessary to determine if this is an acceptable and effective mode of treatment. PMID- 12751938 TI - Thrombus characterization with intravascular ultrasound: potential to predict successful thrombolysis. AB - PURPOSE: To define analysis methods using radiofrequency (RF) data from intravascular ultrasound (IVUS) to characterize thrombus so that the success or failure of thrombolysis can be predicted. METHODS: Experimentation was done in 2 phases: first, 40 clots created from platelet-rich (n=20) and platelet-poor (n=20) plasma were imaged with 20 and 30-MHz IVUS probes. The digitized RF signals were analyzed to determine the attenuation and slope of attenuation characteristics as a reference standard for the second part of the study. In this phase, 20 perfusion trials were performed with 10 platelet-rich and 10 platelet poor samples in an in vitro arterial perfusion model; the composition of the samples was blinded during interrogation with IVUS. Alteplase (0.1 mg) was then infused directly into the thrombus for 1 hour; restoration of flow and amount of lysis were quantified. RESULTS: Significant differences were detected for the integrated attenuation value (p<0.001) between platelet-poor and platelet-rich clots interrogated with a 30-MHz probe, but no differences were noted using the 20-MHz catheter. However, both transducers were capable of differentiating platelet-rich and platelet-poor clots using a slope of attenuation analysis, which was successful in identifying all 20 (100%) of the blinded clots in the in vitro model. The speed and completeness of lysis were significantly higher (p<0.05) for platelet-rich (78%) than platelet-poor (21%) clots. CONCLUSIONS: IVUS preprocessed imaging is capable of differentiating various types of thrombus. This information can be used to predict the success or failure of pharmacological lysis techniques. PMID- 12751939 TI - Suspected pulmonary artery disruption after transvenous pulmonary embolectomy using a hydrodynamic thrombectomy device: clinical case and experimental study on porcine lung explants. AB - PURPOSE: To use porcine lung explants for reconstructing possible situations in which a vessel wall disruption might have occurred in a patient suffering fatal hemoptysis after pulmonary embolectomy with a hydrodynamic thrombectomy device. METHODS: A 76-year-old woman with massive pulmonary embolism underwent transvenous pulmonary embolectomy using a 6-F AngioJet Xpeedior catheter according to manufacturer's instructions. While activating the device in the middle lobe artery (approximately 8 mm diameter), massive and ultimately fatal arterial bleeding occurred through the tracheal tube. Because no autopsy was authorized, an experimental study was designed to examine possible causes for the vessel disruption. Five fresh porcine heart-lung preparations were examined inside a dedicated chest phantom. Access to the pulmonary vessels was provided through catheters inside the right and left ventricular outlets. A low-flow circulation was maintained with an external pump. The 6-F AngioJet thrombectomy device was activated at 42 sites inside vessels from 2 to 10 mm in diameter; in one lung, 8 activations were made after deliberately withdrawing the guidewire. RESULTS: Vessels >6 mm in diameter remained intact. Vessel wall disruption occurred in 4 of 7 vessels between 4 and 6 mm in diameter and in 13 of 14 segmental arteries <4 mm in diameter (regardless of whether or not a guidewire was used). The signs of vessel wall disruption included extravasation of contrast material, arteriovenous fistula, and laceration of distal airspaces with contrast inside the bronchus. CONCLUSIONS: The application of this system has to be considered potentially dangerous when activated inside vessels with diameters <6 mm. The use of this device appears to be safe only inside main branches of the lung vessels at this time. Additional experiments will be required to substantiate these initial results. PMID- 12751940 TI - Treatment of popliteal artery aneurysms with the Hemobahn stent-graft. AB - PURPOSE: To report a prospective study to ascertain the results of popliteal aneurysm treatment with a self-expanding stent-graft. METHODS: In a recent 3-year period, 21 patients (18 men; median age 67 years, range 52-82) with 23 popliteal aneurysms were treated with Hemobahn stent-grafts. Follow-up evaluation included duplex scanning, ankle-brachial index (ABI) measurements, and radiographic examination of the knee. Outcome measures were occlusion of the stent-graft and limb loss. RESULTS: Technical success in placing the stent-graft and excluding the aneurysm was 100%. An additional vascular intervention was performed in the same session in 5 (24%) cases. In the other 16 patients, local anesthesia was used in 10 (63%). During a median follow-up of 15 months (range 2-37), 5 (22%) of 23 stent-grafts occluded, resulting in a cumulative patency of 74%. All occlusions occurred within 6 months after the intervention; 2 were successfully recanalized, and none of the 3 patients with persisting occlusion required an amputation. CONCLUSIONS: The results of this study suggest that endovascular stent-graft repair of popliteal artery aneurysms is feasible. Midterm patency rates are lower compared to traditional surgical repair. PMID- 12751941 TI - Open surgical versus minimally invasive in situ femorodistal bypass: long-term results. AB - PURPOSE: To compare long-term patency and limb survival rates for the classical in situ surgical bypass procedure versus a minimally invasive technique for femorodistal revascularization. METHODS: From May 1992 to June 1994, a prospective multicenter study was undertaken at 4 centers to evaluate the open versus closed technique for femorodistal bypass grafting. Of 97 patients enrolled in the trial, 73 patients (49 men; mean age 71 years) were assigned to the long term follow-up protocol and prospectively randomized to the open (n=38) or closed (n=35) procedure. The classical open technique is characterized by a long incision over the length of the bypass graft, while the minimally invasive procedure involves only two short incisions over each anastomosis site (the side branches are closed with a coaxial embolization catheter system). Graft patency was evaluated with duplex imaging periodically throughout the 4-year observation period. RESULTS: There was no statistically significant difference between the treatment groups with respect to age, sex, hypertension, ischemic heart disease, or smoking. However, the open group had a significantly greater incidence of diabetes (p=0.037). Over a median 4.7-year follow-up (range 0.3-6.4), 9 (12%) patients (3 open and 6 closed) were lost to follow-up: 2 died and 7 refused the duplex examination. No significant differences in 4-year patency, limb salvage, or survival was demonstrated between the open versus closed treatment groups; 4 year secondary patency was 62% versus 64%, respectively, and limb salvage was 72% versus 86%. CONCLUSIONS: The closed technique for femorodistal in situ bypass procedures yields favorable long-term outcomes compared to the traditional open technique. PMID- 12751942 TI - Remote superficial femoral artery endarterectomy and distal vein bypass for limb salvage: initial experience. AB - PURPOSE: To examine the results of remote superficial femoral artery endarterectomy (RSFAE) performed through a small groin incision in conjunction with distal saphenous vein (SV) bypass for limb salvage. METHODS: A retrospective study was conducted of 21 patients (14 men; mean age 68.5 years, range 47-78) who underwent RSFAE and distal SV bypass between May 1998 and September 2001 for limb salvage. Thirteen had gangrene and 8 had rest pain. RSFAE was performed with the MollRing Cutter device through a femoral arteriotomy; the distal atheromatous plaque was "tacked up" with a stent. Distal SV bypass from the proximal popliteal artery was performed in situ in 7, from a transposed harvested vein in 8, or from a reversed graft in 6. All patients underwent follow-up examination with serial color-flow ultrasound scans. RESULTS: The mean length of the endarterectomized SFA was 26.5 cm (range 12-40). There were no deaths, only 2 wound complications, and the mean hospital length of stay was 3.1+/-0.6 days. The primary cumulative patency rate by life-table analysis was 71.4% with follow-up extending to an average of 12.4 months (range 1-18). There were 2 amputations for gangrene and 6 percutaneous procedures in 4 (19.1%) patients to maintain bypass patency, producing an assisted primary patency rate of 81.5%. The locations of the restenoses were evenly distributed along the endarterectomized SFA and SV graft. CONCLUSIONS: When adequate SV is not available, RSFAE with residual SV bypass is a safe and moderately durable procedure that may prove to be a useful adjunct for limb salvage, especially in the presence of foot infection, where an autogenous tissue bypass is preferred. PMID- 12751943 TI - Simple technique to decrease total clamping time during combined iliac and femoral endovascular procedures. AB - PURPOSE: To describe an alternative method of gaining vascular access while preserving blood flow to the profunda femoral artery during combined iliac and superficial femoral artery (SFA) recanalization. TECHNIQUE: A sheath is introduced through the occluded SFA to reduce the total ischemic time when both the external iliac artery and SFA are obstructed. After iliac stenting and remote endarterectomy of the SFA with stent-graft deployment, blood flow to the profunda is interrupted only briefly when the proximal anastomosis of the femoral endobypass is completed. CONCLUSIONS: Sheath insertion into the proximal portion of an occluded SFA can be an effective option in reducing ischemic time when dealing with combined iliac and femoral endovascular procedures. PMID- 12751944 TI - Aortoenteric fistula due to endoleak coil embolization after endovascular AAA repair. AB - PURPOSE: To report a late complication associated with embolization coils used to treat an endoleak after endovascular abdominal aortic aneurysm (AAA) repair. CASE REPORT: A 79-year-old man with a 5.8-cm AAA underwent endovascular repair with an Ancure graft in 1997. A persistent type I endoleak was identified on serial postoperative computed tomographic scans. Three transarterial coil embolization procedures were performed to treat an endoleak from the proximal and right distal attachment sites with outflow by the inferior mesenteric and lumbar arteries. Coil embolization was ultimately successful in sealing the endoleak, and the AAA decreased in size. Four years later, the patient developed an aortoenteric fistula due to erosion of the metallic embolization coils into the duodenum. The endograft was explanted and an extra-anatomical bypass inserted. CONCLUSIONS: Coil embolization to treat endoleaks can, on rare occasions, be the cause of aortoenteric fistula. Lifelong follow-up of stent-graft patients is required. PMID- 12751945 TI - Endovascular aortic biopsy in the diagnosis of takayasu arteritis. AB - PURPOSE: To use a minimally invasive approach to obtain biopsy specimens of the aorta to confirm the diagnosis of Takayasu arteritis and formulate a therapeutic plan. CASE REPORT: A 17-year-old woman with lower extremity claudication and a normal erythrocyte sedimentation rate had magnetic resonance angiography, which suggested stenotic lesions of the infrarenal aorta and left renal artery. An endovascular biopsy of the aortic lesion was performed under local anesthesia, retrieving sufficient tissue to make the diagnosis of Takayasu arteritis. The patient was discharged the same day. CONCLUSIONS: Endovascular aortic biopsy may be useful in diagnosing Takayasu arteritis and guiding the clinician toward the proper mode of treatment. PMID- 12751946 TI - Stenting for pulmonary artery stenosis due to a recurrent primary leiomyosarcoma. AB - PURPOSE: To report stent implantation for a malignant obstruction within the pulmonary artery (PA) caused by a recurrent leiomyosarcoma in the pulmonary trunk. CASE REPORT: A 62-year-old man with a non-metastatic primary leiomyosarcoma of the right PA underwent pneumectomy of the right lung and postoperative radiotherapy in 1994. Six years later, he presented with symptoms of progressive right ventricular dysfunction. Computed tomography (CT) identified a high-grade stenosis of the left PA due to recurrent tumor within the pulmonary trunk extending into the left PA. Transthoracic ultrasound documented severe pulmonary hypertension with a high pressure gradient across the stenosis. A stent was deployed percutaneously, successfully establishing PA patency. Pressure measurements showed a significantly reduced gradient across the stented area. In follow-up, the patient reported subjective improvement of symptoms; CT scans revealed a fully patent stent. His status remained stable 11 months after stent implantation. CONCLUSIONS: PA leiomyosarcoma is a rare and highly malignant tumor. In most cases, surgery can only prolong survival for the short term. Palliative interventional PA stenting performed under local anesthesia can offer improvement in quality of life by reducing excessive pulmonary hypertension. PMID- 12751947 TI - Intrahepatic aneurysmal portohepatic venous shunt: embolization with a tissue adhesive solution. AB - PURPOSE: To report the endovascular treatment of an intrahepatic portosystemic venous shunt (IPSVS) using a tissue adhesive solution. CASE REPORT: A 40-year-old woman without diagnosed liver disease presented for evaluation of vague abdominal pains. A latent portosystemic encephalopathy was diagnosed only during hospitalization, when ultrasound examination disclosed a venous-venous right hepatic communication. The patient was treated with selective percutaneous embolization using a tissue adhesive after blocking the lesion's venous inflow and outflow tracts. The result was excellent, and the patient recovered completely in a few days. After 5 months, the patient is asymptomatic and event free. CONCLUSIONS: Large IPSVS may be successfully treated percutaneously using a tissue adhesive solution, avoiding a more invasive procedure. PMID- 12751948 TI - Deep circumflex iliac artery as a cause of type II endoleak. AB - PURPOSE: To report a collateral pathway involving the deep circumflex iliac artery causing a type II endoleak following endoluminal exclusion of an abdominal aortic aneurysm (AAA). CASE REPORT: A 75-year-old man was investigated for a persistent type II endoleak 2 years after endovascular AAA repair with a Zenith Trifab stent-graft. Angiography revealed contrast in the sac from a lumbar artery fed via a collateral of the deep circumflex iliac artery. The lumbar artery was embolized with coils, but an endoleak persisted and is being followed. CONCLUSIONS: This collateral pathway is easily missed during angiography for endoleaks and should be considered where an endoleak is suspected but cannot be found. PMID- 12751949 TI - Percutaneous treatment of placenta percreta using coil embolization. AB - PURPOSE: To report the use of embolotherapy to avoid hysterectomy in rare placenta percreta. CASE REPORT: A pregnant 34-year-old woman (gravida 3, para 2) was admitted with premature rupture of membranes and vaginal bleeding in the 32nd week. Prenatal B-mode and Doppler ultrasound revealed marked hypervascularity of the placenta with disruption of the uterine-bladder interface consistent with placenta percreta. Since the patient insisted on uterine preservation, uterus and placenta were left in situ after caesarean section, which was followed by coaxial microcoil embolization of 6 pelvic arteries and postoperative methotrexate administration. Three months later, the patient had severe bleeding from the retained placenta, possibly under the influence of anticoagulation administered for pulmonary embolism. Emergent hysterectomy was performed. CONCLUSIONS: Coil embolization may avoid immediate hysterectomy and reduce peri-delivery blood loss in placenta percreta. However, retained placenta poses a serious risk, even after months, and secondary hysterectomy should be performed as an elective procedure after embolization. PMID- 12751950 TI - Spontaneous common iliac artery dissection after exercise. AB - PURPOSE: To review the clinical features and management of spontaneous iliac dissections. CASE REPORT: A healthy 60-year-old competitive cyclist presented with acute onset of short-distance claudication following vigorous exercise. Angiography showed a dissection flap extending from the right common iliac artery to the external iliac artery. An uncovered stent was placed across the proximal entry site but did not obliterate the false lumen; open surgical intervention was required. CONCLUSIONS: Spontaneous dissection of the iliac artery is a rare but important condition to suspect in high performance athletes complaining of leg pain following exercise. PMID- 12751951 TI - Training in the radiation sciences: a national and international need. PMID- 12751952 TI - Computerized video time-lapse (CVTL) analysis of the fate of giant cells produced by X-irradiating EJ30 human bladder carcinoma cells. AB - This study was designed to examine the viability and proliferation of uninucleated and multinucleated giant cells formed after 6 Gy X irradiation. The pedigrees of 102 individual EJ30 giant cells present 5 days after irradiation were analyzed from time-lapse movies captured over 6.3 days from 100 fields (100x). Pedigree analysis enabled us to study the proliferation of giant cells. The average starting size (area) of the giant cells (14500 +/- 9100 microm(2)) was approximately 25 times larger than the normal-sized cells observed after irradiation (560 +/- 200 microm(2)). From a total of 76 pedigrees of uninucleated giant cells, 42 had giant cells that either died or were arrested, while 34 divided at least once and produced progeny that divided again (five three times and three four times) before the progeny died or were arrested. Twenty-four pedigrees contained progeny that were lost from observation after dividing at least once. While most progeny continued to have giant cell morphology, two uninucleated giant cells ultimately produced progeny that contained two normal sized cells. From a total of 26 multinucleated giant cells, only three divided. Two divided only once, but one produced progeny that divided two times. In all, 37 out of 102 giant cells divided at least once; eight of these divided four or five times with two of these pedigrees ultimately producing two normal-sized daughter cells. These results suggest that a small fraction of giant cells might be potentially clonogenic. PMID- 12751953 TI - Radiation-induced micronucleus frequency in peripheral blood lymphocytes is correlated with normal tissue damage in patients with cervical carcinoma undergoing radiotherapy. AB - In an effort to find a test to predict the response of normal tissue to radiotherapy, the lymphocyte micronucleus assay was used on blood samples from patients with cervical carcinoma. Peripheral blood samples from 55 patients with advanced-stage (II B-IV B) cervical carcinoma were obtained before radiotherapy. The patients were treated with external-beam radiotherapy followed by high-dose rate brachytherapy. Acute and late normal tissue reactions were scored and correlated with the micronucleus frequency in lymphocytes after irradiation with 4 Gy in vitro. Great interindividual variability was observed in the radiation induced lymphocyte micronucleus frequency, especially at 4 Gy. The mean number of micronuclei per 100 binucleated cells in cells irradiated with 4 Gy in vitro was significantly higher in samples from patients who suffered from acute and/or late normal tissue reactions than in those from patients with no reactions (51.0 +/- 17.7 and 29.6 +/- 10.1, respectively). A significant correlation was also found between the micronucleus frequency at 4 Gy and the severity of acute reactions and late reactions. However, the overlap between the micronucleus frequencies of patients with high-grade late normal tissue reactions and low-grade reactions is too great to recommend the micronucleus assay in its present form for routine clinical application. PMID- 12751954 TI - Radiation-dependent limit for the viability of bacterial spores in halite fluid inclusions and on Mars. AB - When claims for the long-term survival of viable organisms are made, either within terrestrial minerals or on Mars, considerations should be made of the limitations imposed by the naturally occurring radiation dose to which they have been exposed. We investigated the effect of ionizing radiation on different bacterial spores by measuring the inactivation constants for B. subtilis and S. marismortui spores in solution as well as for dry spores of B. subtilis and B. thuringiensis. S. marismortui is a halophilic spore that is genetically similar to the recently discovered 2-9-3 bacterium from a halite fluid inclusion, claimed to be 250 million years old (Vreeland et al., Nature 407, 897-900, 2000). B. thuringiensis is a soil bacterium that is genetically similar to the human pathogens B. anthracis and B. cereus (Helgason et al., Appl. Environ. Microbiol. 66, 2627-2630, 2000). To relate the inactivation constant to some realistic environments, we calculated the radiation regimen in a halite fluid inclusion and in the Martian subsurface over time. Our conclusion is that the ionizing dose of radiation in those environments limits the survival of viable bacterial spores over long periods. In the absence of an active repair mechanism in the dormant state, the long-term survival of spores is limited to less than 109 million years in halite fluid inclusions, to 100 to 160 million years in the Martian subsurface below 3 m, and to less than 600,000 years in the uppermost meter of Mars. PMID- 12751955 TI - The TP53 dependence of radiation-induced chromosome instability in human lymphoblastoid cells. AB - The dose and TP53 dependence for the induction of chromosome instability were examined in cells of three human lymphoblastoid cell lines derived from WIL2 cells: TK6, a TP53-normal cell line, NH32, a TP53-knockout created from TK6, and WTK1, a WIL2-derived cell line that spontaneously developed a TP53 mutation. Cells of each cell line were exposed to (137)Cs gamma rays, and then surviving clones were isolated and expanded in culture for approximately 35 generations before the frequency and characteristics of the instability were analyzed. The presence of dicentric chromosomes, formed by end-to-end fusions, served as a marker of chromosomal instability. Unexposed TK6 cells had low levels of chromosomal instability (0.002 +/- 0.001 dicentrics/cell). Exposure of TK6 cells to doses as low as 5 cGy gamma rays increased chromosome instability levels nearly 10-fold to 0.019 +/- 0.008 dicentrics/cell. There was no further increase in instability levels beyond 5 cGy. In contrast to TK6 cells, unexposed cultures of WTK1 and NH32 cells had much higher levels of chromosome instability of 0.034 +/- 0.007 and 0.041 +/- 0.009, respectively, but showed little if any effect of radiation on levels of chromosome instability. The results suggest that radiation exposure alters the normal TP53-dependent cell cycle checkpoint controls that recognize alterations in telomere structure and activate apoptosis. PMID- 12751956 TI - Induction of genomic instability in TK6 human lymphoblasts exposed to 137Cs gamma radiation: comparison to the induction by exposure to accelerated 56Fe particles. AB - The induction of genomic instability in TK6 human lymphoblasts by exposure to (137)Cs gamma radiation was investigated by measuring the frequency and characteristics of unstable clones isolated approximately 36 generations after exposure. Clones surviving irradiation and control clones were analyzed for 17 characteristics including chromosomal aberrations, growth defects, alterations in response to a second irradiation, and mutant frequencies at the thymidine kinase and Na(+)/K(+) ATPase loci. Putative unstable clones were defined as those that exhibited a significant alteration in one or more characteristics compared to the controls. The frequency and characteristics of the unstable clones were compared in clones exposed to (137)Cs gamma rays or (56)Fe particles. The majority of the unstable clones isolated after exposure to either gamma rays or (56)Fe particles exhibited chromosomal instability. Alterations in growth characteristics, radiation response and mutant frequencies occurred much less often than cytogenetic alterations in these unstable clones. The frequency and complexity of the unstable clones were greater after exposure to (56)Fe particles than to gamma rays. Unstable clones that survived 36 generations after exposure to gamma rays exhibited increases in the incidence of dicentric chromosomes but not of chromatid breaks, whereas unstable clones that survived 36 generations after exposure to (56)Fe particles exhibited increases in both chromatid and chromosome aberrations. PMID- 12751957 TI - Depletion of KIN17, a human DNA replication protein, increases the radiosensitivity of RKO cells. AB - The human KIN17 protein is a chromatin-associated protein involved in DNA replication. Certain tumor cell lines overproduce KIN17 protein. Among 16 cell lines, the highest KIN17 protein level was observed in H1299 non-small cell lung cancer cells, whereas the lowest was detected in MeWo melanoma cells. Cells displaying higher KIN17 protein levels exhibited elevated RPA70 protein contents. High KIN17 protein levels may be a consequence of the tumorigenic phenotype or a prerequisite for tumor progression. Twenty-four hours after exposure to ionizing radiation, after the completion of DNA repair, a co-induction of chromatin-bound KIN17 and RPA70 proteins was detected. Etoposide, an inhibitor of topoisomerase II generating double-strand breaks, triggered the concentration of KIN17 into punctuate intranuclear foci. KIN17 may be associated with unrepaired DNA sites. Flow cytometry analysis revealed that 48 h after transfection the uppermost KIN17 positive RKO cells shifted in the cell cycle toward higher DNA content, suggesting that KIN17 protein induced defects in chromatin conformation. Cells displaying reduced levels of KIN17 transcript exhibited a sixfold increased radiosensitivity at 2 Gy. The KIN17 protein may be a component of the DNA replication machinery that participates in the cellular response to unrepaired DSBs, and an impaired KIN17 pathway leads to an increased sensitivity to ionizing radiation. PMID- 12751958 TI - Cell cycle-dependent expression of phosphorylated histone H2AX: reduced expression in unirradiated but not X-irradiated G1-phase cells. AB - Exposure of cells to ionizing radiation causes phosphorylation of histone H2AX at sites flanking DNA double-strand breaks. Detection of phosphorylated H2AX (gammaH2AX) by antibody binding has been used as a method to identify double strand breaks. Although generally performed by observing microscopic foci within cells, flow cytometry offers the advantage of measuring changes in gammaH2AX intensity in relation to cell cycle position. The importance of cell cycle position on the levels of endogenous and radiation-induced gammaH2AX was examined in cell lines that varied in DNA content, cell cycle distribution, and kinase activity. Bivariate analysis of gammaH2AX expression relative to DNA content and synchronization by centrifugal elutriation were used to measure cell cycle specific expression of gammaH2AX. With the exception of xrs5 cells, gammaH2AX level was approximately 3 times lower in unirradiated G(1)-phase cells than S- and G(2)-phase cells, and the slope of the G(1)-phase dose-response curve was 2.8 times larger than the slope for S-phase cells. Cell cycle differences were confirmed using immunoblotting, indicating that reduced antibody accessibility in intact cells was not responsible for the reduced antibody binding in G(1)-phase cells. Early apoptotic cells could be easily identified on flow histograms as a population with 5-10-fold higher levels of gammaH2AX, although high expression was not maintained in apoptotic cells by 24 h. We conclude that expression of gammaH2AX is associated with DNA replication in unirradiated cells and that this reduces the sensitivity for detecting radiation-induced double-strand breaks in S and G(2)-phase cells. PMID- 12751959 TI - Radioprotective effects of ginsan, an immunomodulator. AB - We previously reported that ginsan, a purified polysaccharide isolated from Panax ginseng, had a mitogenic activity, induced LAK cells, and increased levels of several cytokines. In an effort to identify other immunostimulatory effects, we evaluated the protective effects of ginsan injected in vivo against radiation by measuring its effects on the CFU-S bone marrow cells and spleen cells. Ginsan was found to significantly increase the number of bone marrow cells, spleen cells, granulocyte-macrophage colony-forming cells (GM-CFC), and circulating neutrophils, lymphocytes and platelets in irradiated mice. In addition, ginsan induced the endogenous production of cytokines such as Il1, Il6, Ifng and Il12, which are required for hematopoietic recovery, and was able to enhance Th1 function while interfering with the Th2 response in irradiated mice. We demonstrated that pretreatment with ginsan protected mice from the lethal effects of ionizing radiation more effectively than when it was given immediately after or at various times after irradiation. A significant increase in the LD(50/30) from 7.54 Gy for PBS injection to 10.93 Gy for mice pretreated with 100 mg/kg ginsan was observed. These findings indicate that ginsan may be a useful agent to reduce the time necessary for reconstituting hematopoietic cells after irradiation. PMID- 12751960 TI - Effects of 10-T static magnetic field on human peripheral blood immune cells. AB - The exposure of peripheral blood mononuclear cells (PBMCs) was performed in a 10 T static magnetic field. Without lymphocyte stimulation, there were no significant differences in the viability of the exposed and unexposed CD4(+) T cells, CD8(+) T cells, B cells, and natural killer (NK) cells. The expression of Th1 type chemokine receptor, CXCR3, and Th2 type receptor, CCR3, was unaltered after magnetic-field exposure. No differences were observed in the naive T cells and memory T-cell subclasses in either CD4(+) or CD8(+) T cells. In contrast to the unstimulated condition, the magnetic-field exposure reduced the viability of phytohemagglutinin (PHA)-activated T cells in both the CD4(+) and CD8(+) subclasses. In particular, the number of PHA-treated naive CD8(+) T cells (CD45RA(+)CD4(-)CD8(+)) was markedly decreased after the magnetic-field exposure, while PHA-treated memory CD8(+) cells (CD45RA(-)CD4(-)CD8(+)) were resistant to the exposure. The number of PHA-treated naive CD4(+) T cells (CD45RA(+)CD4(+)CD8( )) and memory cells (CD45RA(-)CD4(+)CD8(-)) was markedly decreased to a similar degree. Thus the susceptibility of lymphocytes to the magnetic-field exposure differed among activated T-cell subtypes. The magnetic-field exposure significantly increased the death of PHA-stimulated lymphocytes by apoptosis. These results suggest that a strong static magnetic field has acute effects on immune cells during cell division, while the field exposure has a minimal effect on immune cells in a nondividing phase. PMID- 12751961 TI - Prevalence of hepatitis B virus infection among atomic bomb survivors. AB - The aim of this study was to determine whether the prevalence of hepatitis B virus (HBV) carriers increased with atomic bomb radiation dose, and whether radiation decreased the ability to clear HBV among the atomic bomb survivors. The study subjects were 6,121 participants in the Adult Health Study of atomic bomb survivors in Hiroshima and Nagasaki. After adjustment for age, sex, city and potential confounders, the rates of seropositivity for hepatitis B surface antigen (HBsAg), indicating current HBV infections, and anti-hepatitis B core antibody, indicating either cured or current infections, increased with radiation dose. However, no relationship was observed between radiation and anti-hepatitis B surface antibody (indicating cured infection). The proportion of persons who were unable to clear the virus, as the proportion of HBsAg-positive persons among those ever infected by HBV (positive for HBsAg or surface or core hepatitis B antibody), increased significantly with radiation dose among those receiving blood transfusions. This proportion was not related to dose among those who reported no such transfusions. The findings may suggest a lower likelihood of clearance after HBV infection among those who were more likely to have been infected with HBV as adults after atomic bomb irradiation rather than as infants or adults prior to irradiation. PMID- 12751962 TI - Cancer mortality risk among workers at the Mayak nuclear complex. AB - At present, direct data on risk from protracted or fractionated radiation exposure at low dose rates have been limited largely to studies of populations exposed to low cumulative doses with resulting low statistical power. We evaluated the cancer risks associated with protracted exposure to external whole body gamma radiation at high cumulative doses (the average dose is 0.8 Gy and the highest doses exceed 10 Gy) in Russian nuclear workers. Cancer deaths in a cohort of about 21,500 nuclear workers who began working at the Mayak complex between 1948 and 1972 were ascertained from death certificates and autopsy reports with follow-up through December 1997. Excess relative risk models were used to estimate solid cancer and leukemia risks associated with external gamma-radiation dose with adjustment for effects of plutonium exposures. Both solid cancer and leukemia death rates increased significantly with increasing gamma-ray dose (P < 0.001). Under a linear dose-response model, the excess relative risk for lung, liver and skeletal cancers as a group (668 deaths) adjusted for plutonium exposure is 0.30 per gray (P < 0.001) and 0.08 per gray (P < 0.001) for all other solid cancers (1062 deaths). The solid cancer dose-response functions appear to be nonlinear, with the excess risk estimates at doses of less than 3 Gy being about twice those predicted by the linear model. Plutonium exposure was associated with increased risks both for lung, liver and skeletal cancers (the sites of primary plutonium deposition) and for other solid cancers as a group. A significant dose response, with no indication of plutonium exposure effects, was found for leukemia. Excess risks for leukemia exhibited a significant dependence on the time since the dose was received. For doses received within 3 to 5 years of death the excess relative risk per gray was estimated to be about 7 (P < 0.001), but this risk was only 0.45 (P = 0.02) for doses received 5 to 45 years prior to death. External gamma-ray exposures significantly increased risks of both solid cancers and leukemia in this large cohort of men and women with occupational radiation exposures. Risks at doses of less than 1 Gy may be slightly lower than those seen for doses arising from acute exposures in the atomic bomb survivors. As dose estimates for the Mayak workers are improved, it should be possible to obtain more precise estimates of solid cancer and leukemia risks from protracted external radiation exposure in this cohort. PMID- 12751963 TI - Additive effects of radiation and docetaxel on murine SCCVII tumors in vivo: special reference to changes in the cell cycle. AB - The purpose of the present study was to investigate the effects of a combination of docetaxel and irradiation in vivo with special reference to docetaxel-arrested G(2)/M-phase cells. At 24 and 48 h after intraperitoneal administration of docetaxel (90 mg/kg), tumor-bearing mice were irradiated with (60)Co gamma rays. Cell cycle distribution was analyzed by a DNA-Ki-67 double staining method using flow cytometry. An accumulation of cells in the G(2)/M phase of up to approximately 40% was observed 24 h after administration of docetaxel. Between 24 and 72 h, the percentage of cells arrested in G(2)/M phase that expressed Ki-67 decreased from 37.2% to 13.8%, in accordance with the increase in the Ki-67 negative G(2)/M-phase fraction. More than half of the cells arrested in G(2)/M phase lost their expression of Ki-67 protein between 24 and 72 h. The G(1)-phase fraction decreased from 28.4% to 8.6% at 24 h after docetaxel treatment; this remained unchanged at 72 h. These flow cytometry data suggested that docetaxel arrested G(2)/M-phase cells did not enter the next cell cycle and were killed by docetaxel alone. Our data showed that arrest of cells in G(2)/M phase does not contribute to the synergism that has been reported for combinations of docetaxel and radiation in in vivo tumor models. PMID- 12751964 TI - Increased binding and chemotactic capacities of PDGF-BB on fibroblasts in radiation pneumonitis. AB - Although pulmonary fibrosis is a frequent and serious consequence of radiotherapy for thoracic malignant diseases such as lung cancer, the pathogenesis of this radiation-induced lung disorder remains unclear. To clarify the mechanisms underlying radiation pneumonitis and pulmonary fibrosis, we investigated the expression of platelet-derived growth factor receptor (PDGFR) on fibroblasts obtained from irradiated rat lungs and on control fibroblasts. Whole lungs of male Wistar rats were irradiated with a single dose of 15 Gy, and lung fibroblasts were isolated at 4 weeks after the irradiation. The chemotactic response of irradiated lung fibroblasts to PDGF-BB was significantly higher than that of control lung fibroblasts, whereas there was no significant difference between irradiated lung fibroblasts and control lung fibroblasts in the response to PDGF-AA. Receptor binding assay showed more specific binding sites for PDGF-BB on irradiated lung fibroblasts than on control lung fibroblasts, and the displacement of (125)I-labeled PDGF binding to fibroblasts by unlabeled PDGF showed that (125)I-labeled PDGF-BB was displaced by PDGF-BB but not by PDGF-AA. These results suggest that the increased binding sites for PDGF-BB on irradiated lung fibroblasts correspond mainly to PDGFRB. Scatchard analysis of the saturation data demonstrated an approximately twofold increase both in the number of PDGF-BB binding sites and in the binding affinity in irradiated lung fibroblasts compared to that in control lung fibroblasts. Those results suggest that the increased chemotactic response of irradiated lung fibroblasts to PDGF-BB is related to the overexpression of PDGFRB, which may have an important role in the pathogenesis of radiation-induced pneumonitis and pulmonary fibrosis. PMID- 12751965 TI - Molecular and cellular biology of moderate-dose (1-10 Gy) radiation and potential mechanisms of radiation protection: report of a workshop at Bethesda, Maryland, December 17-18, 2001. AB - Exposures to doses of radiation of 1-10 Gy, defined in this workshop as moderate dose radiation, may occur during the course of radiation therapy or as the result of radiation accidents or nuclear/radiological terrorism alone or in conjunction with bioterrorism. The resulting radiation injuries would be due to a series of molecular, cellular, tissue and whole-animal processes. To address the status of research on these issues, a broad-based workshop was convened. The specific recommendations were: (1) RESEARCH: Identify the key molecular, cellular and tissue pathways that lead from the initial molecular lesions to immediate and delayed injury. The latter is a chronic progressive process for which postexposure treatment may be possible. (2) Technology: Develop high-throughput technology for studying gene, protein and other biochemical expression after radiation exposure, and cytogenetic markers of radiation exposure employing rapid and accurate techniques for analyzing multiple samples. (3) Treatment strategies: Identify additional biological targets and develop effective treatments for radiation injury. (4) Ensuring sufficient expertise: Recruit and train investigators from such fields as radiation biology, cancer biology, molecular biology, cellular biology and wound healing, and encourage collaboration on interdisciplinary research on the mechanisms and treatment of radiation injury. Communicate knowledge of the effects of radiation exposure to the general public and to investigators, policy makers and agencies involved in response to nuclear accidents/events and protection/treatment of the general public. PMID- 12751966 TI - Further comments on "Long-term exposure of Emu-Pim1 transgenic mice to 898.4 MHz microwaves does not increase lymphoma incidence" by Utteridge et al. (Radiat. Res. 158, 357-364 2002). PMID- 12751968 TI - You've come a long way baby: Women and the tobacco epidemic. PMID- 12751969 TI - Socioeconomic status or social contexts. PMID- 12751970 TI - Alexander the Great's relationship with alcohol. AB - AIMS: This study sought to clarify if Alexander the Great indulged pathologically in alcohol and whether it contributed to his death. DESIGN: The texts of the historians Diodorus of Sicily, Plutarch, Arrian, Curtius Rufus, Athenaeus, Aelian and Justin were studied, with their information concerning wine consumption by Macedonians, and especially Alexander, and were evaluated. The surviving historical texts, all later than Alexander's epoch, are based on a series of contemporary histories and especially on the 'Royal Journals', an official diary written in the imperial court. FINDINGS AND CONCLUSIONS: Alexander consumed large quantities of undiluted wine periodically, reaching pathological intoxication. However, the existing data do not provide convincing evidence that Alexander the Great manifested abuse of or dependence on alcohol according to DSM-IV or ICD-10 criteria and it seems unlikely that alcohol was involved in his untimely death. PMID- 12751971 TI - Treatments for spit tobacco use: a quantitative systematic review. AB - AIMS: Spit tobacco use is prevalent in the United States and is associated with adverse health consequences. Health-care providers have neither evidence summaries nor evidence-based guidelines to assist them in treating patients who use spit tobacco. DESIGN: We completed a systematic review of the literature to determine the efficacy and safety of pharmacological and behavioral interventions for the treatment of spit tobacco use. FINDINGS: We found six randomized controlled trials testing pharmacological interventions and eight testing behavioral interventions. Using random-effects meta-analyses,bupropion sustained release (SR) increased point prevalence tobacco abstinence at 12 weeks [odds ratio (OR) 2.1; 95% confidence interval (CI), 1.0-4.2]. Nicotine replacement therapy with patch or gum increased point prevalence tobacco abstinence at 6 months (OR 1.3; 95% CI, 1.0-1.6). Behavioral interventions increased long-term (6 month)point prevalence tobacco abstinence (OR 1.7; 95% CI, 1.1-2.9). Studies including an oral examination followed by feedback to the patient had the highest treatment effect. CONCLUSIONS: Behavioral interventions for ST users are effective for increasing ST abstinence rates. Bupropion SR is probably effective and nicotine replacement therapy may be effective. This evidence from randomized controlled trials provides health-care professionals with information necessary to effectively treat spit tobacco use. PMID- 12751972 TI - Nine-year prospective relationship between parental smoking cessation and children's daily smoking. AB - AIMS: The first prospective investigation of the extent to which parental smoking cessation predicts their children's daily smoking. DESIGN: Parental smoking status was assessed when children were aged 8/9 years and children's smoking status was assessed at age 17/18 years. SETTING: Twenty Washington State school districts in the control group of the Hutchinson Smoking Prevention Project. PARTICIPANTS AND MEASUREMENTS: Questionnaire data were gathered on 3012 children (49% female and 91% Caucasian) and both of their parents in a cohort with a 95% retention rate. FINDINGS: When both parents quit smoking, children's odds of daily smoking were reduced by 39% (95% CI = 15%,56%) compared to when both parents were current smokers.Furthermore, when both parents never smoked then children's odds of daily smoking were reduced by 71% (95% CI = 62%,78%). CONCLUSIONS: Parental smoking cessation is associated with reduced risk of their children's daily smoking. Parents who quit still place children at substantially higher risk compared to parents who never smoked. PMID- 12751973 TI - Parental smoking cessation and adolescent smoking: a commentary on Bricker et al. PMID- 12751974 TI - Parental smoking cessation and children's daily smoking: public health implications? commentary on Bricker et al. PMID- 12751975 TI - Parental smoking cessation and children's smoking--problems of interpretation remain: commentary on Bricker et al. PMID- 12751977 TI - Socioeconomic status and drinking patterns in young adults. AB - AIMS: To investigate the relationship between several indicators of socioeconomic status and drinking patterns in young adulthood. DESIGN: Data collected in a longitudinal study of young adults was analysed using repeated-measures models to examine the relationship between income, occupational activity and educational achievement and patterns of drinking. SETTING: These data were collected as part of a longitudinal study ofa birth cohort of New Zealanders. They were interviewed for the most part in a central location using a face-to-face method and a computer-assisted alcohol interview. PARTICIPANTS: The participants were members of the Dunedin Multidisciplinary Health and Development study aged 18, 21 and 26 years. Nine hundred and sixty-nine study members contributed to the analysis. Study members have been found to be broadly representative of the New Zealand population and cross national studies suggest findings are generalizable to other similar market economies. MEASUREMENTS: Three indicators of socioeconomic status were used; educational achievement, occupational activity and income. The educational achievement indicator at age 18 had three levels that ranged from no school qualifications to higher school qualifications. For age 21 two additional categories of tertiary educational achievement were included to make five categories and for age 26 higher tertiary degrees were included in the measure to make six categories. Five categories of occupational activity were used. Income data was also used. Two measures of alcohol consumption were used. These were the frequency of drinking and the typical quantity of alcohol consumed per drinking occasion in the past year. FINDINGS: Frequency of drinking increased over these early adult years and the quantities consumed peaked at age 21 and decreased thereafter for both males and females. Frequency of drinking was influenced by income with the higher income respondents drinking more often and this was persistent overtime. Quantity of drinking was most influenced by educational achievement. The less well-educated young adult drank significantly more during a drinking occasion and at all ages. There was also a relationship between educational achievement and frequency of drinking for males at age 18 and a relationship between women's occupational activity and the quantities they consumed. CONCLUSIONS: The finding that the dimensions of drinking operate differently explains the lack of consistency in previous research, which has investigated socioeconomic status and the volumes of alcohol consumed. The findings of higher quantities consumed among those of lower social status may explain some of the reduced life expectancy found among those with lower socioeconomic status. PMID- 12751978 TI - Alcohol drinking as a predictor of intensive care and hospital mortality in general surgery: a prospective study. AB - AIMS: To analyse whether alcohol drinking increases admission to intensive care and in-hospital mortality in general surgery. DESIGN AND PARTICIPANTS: A prospective cohort study on a consecutive series of 1505 hospitalized patients in a Service of General Surgery of a tertiary hospital. MEASUREMENTS: Drinking pattern was defined by quantity, frequency and volume of drinking. Information on relevant confounders was obtained: smoking, body mass index, nutritional status (measured by serum albumin), cholesterol and its fractions, severity of the underlying disease and all therapeutic measures. Multivariate logistic regression was applied to assess the relationship between drinking and both admission to intensive care and in-hospital death. RESULTS: Twenty-nine (1.9%) patients died and 33 (2.1%) were admitted to the intensive care unit (ICU). Drinking was heavier in men, patients without antecedents of cancer, with lower preoperative risk assessment scores, number of co-morbidities and age and higher serum albumin levels. After adjusting for age, severity of underlying disease, smoking and serum albumin, male drinkers of 72+ g/day had an increased risk of being admitted to ICU, the effect being stronger for week-day drinking (odds ratio, OR = 8.48; 95% confidence interval, CI = 1.68-42.8). A significant association was also seen between week-day drinking (72+ g/day) and death in men (OR = 7.19, 95% CI = 1.43 36.1). Numbers for women were too small to evaluate. CONCLUSION: Heavy drinking increases admission to intensive care and in-hospital mortality in hospitalized male patients undergoing general surgery procedures. PMID- 12751979 TI - Physicians' opinions about medications to treat alcoholism. AB - AIMS: Medications play a limited role in the treatment of alcoholism. This paper examines physicians' opinions about and use of two alcoholism medications currently approved in the US--disulfiram and naltrexone--and one alcoholism medication--acamprosate--that might be approved. DESIGN: A total of 1388 substance abuse specialist physicians who were members of the American Academy of Addiction Psychiatry or the American Society of Addiction Medicine completed a questionnaire in 2001 (65% response rate). FINDINGS: The average percentages of physicians' patients with alcoholism who were prescribed the following medications were: 13% (naltrexone), 9% (disulfiram), 46% (antidepressants) and 11% (benzodiazepines). Almost all physicians had heard of naltrexone and disulfiram, but their self-reported level of knowledge about these medications was lower than for antidepressants. Physicians estimated that naltrexone had a small-to-medium effect size, which was similar in magnitude to the effect size reported in recent meta-analyses of randomized clinical trials. Physicians identified the following three courses of action as the most likely to result in greater use of medications to treat alcohol dependence: more research to develop new medications (33%), more education of physicians about existing medications (17%), and increased involvement of physicians in alcoholism treatment (17%). CONCLUSIONS: Physicians' low rate of use of naltrexone may reflect its small-to medium effect size. PMID- 12751980 TI - Education of key personnel in student pubs leads to a decrease in alcohol consumption among the patrons: a randomized controlled trial. AB - AIMS: To decrease alcohol consumption among patrons in student pubs by server training programmes. DESIGN: Randomized controlled trial. SETTING: University campus. PARTICIPANTS: A total of 1322 students visiting local student pubs during ordinary pub evenings. INTERVENTION: Educational programmes were given to bartenders (n = 40) in a randomized design in six of 12 pubs on a university campus. Bartenders in control pubs were not given the programme. MEASUREMENTS: Breath alcohol concentration (BAC), expressed in percentage, among the patrons and the reported social atmosphere in the pub ('high', 'cosy' and 'rowdy') measured on a visual analogue scale in the pub before and after the intervention programme was given. FINDINGS: BACs of patrons in the intervention pubs were reduced by more than those of the patrons in the control pubs at a 1-month follow up. The mean difference in BAC between intervention and control groups was 0.011% (95% confidence interval, 0.022-0.000). The intervention group also decreased more in reported level of 'rowdy' social atmosphere than did the control group. The mean difference was -6 points (95% confidence interval -11 to 1). No differences were found in reported 'cosy' and 'high' atmosphere. CONCLUSION: Alcohol levels among the patrons were decreased and the 'rowdy' social atmosphere reduced in the intervention group. Server-training programmes for personnel in student pubs could be a component in the prevention of alcohol problems in university student populations. PMID- 12751981 TI - Is there a progression from abuse disorders to dependence disorders? AB - BACKGROUND: Recent studies suggest that a progression occurs from alcohol abuse to alcohol dependence. Although DSM-IV criteria for all substance use-related diagnoses are based largely on the alcohol dependence syndrome, progression from abuse to dependence might not generalize to other substances. AIMS: This study tested whether a progression from DSM-IV abuse to dependence occurs related to the use of cannabis, cocaine and opiates. DESIGN: Retrospective data from the DSM IV Substance Use Disorders Work Group (n = 1226) were reanalysed using McNemar's chi2, configural frequency analyses and survival analyses. Participants were men and women who were primarily African-Americans or Caucasians. SETTINGS: Participants were recruited from community and clinical settings. The measure was the Composite International Diagnostic Interview-Substance Abuse Module. FINDINGS: For all substances, life-time dependence in the absence of life-time abuse was rare. Results were consistent with a progression occurring for alcohol and cannabis, but not for cocaine and opiates. Abuse and dependence occurred in the same year for 66% of the cocaine users who experienced both disorders (57% of users with any cocaine disorder) and 65% of the opiate users who experienced both disorders (46% of users with any opiate disorder). Because cocaine and opiate dependence in the absence of abuse were rare, it is possible that progressions in cocaine and opiate disorders occur more rapidly than cannabis and alcohol. CONCLUSIONS: Research is needed to clarify the mechanisms that influence progressions of substance use disorders. Potential factors leading to between drug variation in rate of progression of disorders are discussed. PMID- 12751982 TI - A comparison of treatment-seeking pathological gamblers based on preferred gambling activity. AB - AIMS: To compare and contrast gamblers with different forms of problematic gambling activities. DESIGN, SETTING AND MEASUREMENTS: Pathological gamblers completed the Addiction Severity Index (ASI) and gambling questionnaires when initiating out-patient treatment. PARTICIPANTS: Participants (n = 347) were categorized by their most problematic form of gambling activity: sports, horse/dog-races, cards, slots and scratch/lottery tickets. Differences in demographics, gambling variables, and ASI composite scores were compared across groups. FINDINGS: After controlling for demographic variables, the types of gamblers differed in severity of gambling, alcohol and psychiatric problems. Horse/dog-race gamblers were generally older, male and less educated; they began gambling regularly at a young age and spent relatively high amounts of money gambling. Sports gamblers were young males and had intermediary gambling problems; they had relatively high rates of current substance use but few psychiatric problems. Card players spent low to moderate amounts of time and money gambling, and they generally reported few alcohol problems and little psychiatric distress. Slot machine players were older and more likely to be female. Slot gamblers began gambling later in life, had high rates of bankruptcy and reported psychiatric difficulties. Scratch/lottery gamblers spent the least amount of money gambling, but they gambled the most frequently and had relatively severe alcohol and psychiatric symptoms. CONCLUSIONS: Gambling patterns and severity of psychosocial problems vary by form of problematic gambling, and these differences may influence treatment recommendations and outcomes. PMID- 12751983 TI - A family history of smoking predicts heightened levels of stress-induced cigarette craving. AB - AIMS: Individuals with histories of smoking in first-degree relatives are significantly more likely to be persistent smokers themselves. The mechanisms underlying this relationship are unknown. Considerable research has demonstrated that smokers display heightened levels of cigarette craving after being exposed to stressful situations, and the magnitude of these craving responses is thought to be predictive of later cessation failure. Based on this research, we tested experimentally the hypothesis that smokers with two or more first-degree relatives who smoked (FH+) would exhibit stronger craving reactions following stressful stimuli than smokers without such family histories (FH-). PARTICIPANTS: We recruited 83 smokers by advertisement (mean age = 41.2 years, 57% female, 41% completed some college, 59% African American). SETTING: The study was conducted in an interview room in an urban medical center. DESIGN: Participants were exposed to a neutral situation (changing a lightbulb) and a stressful situation (dental work) using script-guided imagery. MEASUREMENTS: Participants completed background measures of demographics, distress and smoking behavior. In addition, participants completed cigarette craving and anxiety questionnaires immediately before and after each condition. FINDINGS: Supporting the study hypothesis, FH+ smokers (n = 39) selectively displayed stronger craving reactions to dental imagery (P < 0.03) than did FH- smokers (n = 44). CONCLUSION: The higher levels of stress-induced cigarette craving demonstrated experimentally for individuals with family histories of smoking suggest one mechanism for their poorer cessation success. PMID- 12751984 TI - Effects of reducing contingency management values on heroin and cocaine use for buprenorphine- and desipramine-treated patients. AB - AIMS: During 3 months where contingency management (CM) had an escalating value for each consecutive drug-free urine (escalating CM), cocaine- and heroin-abusing patients significantly increased drug-free urines. The 'escalating CM' was eliminated during months 4-6 to assess any reduction in drug-free urines. DESIGN: Patients who completed a 3-month, randomized, double-blind, trial evaluating CM versus non-CM and desipramine (DMI) versus placebo, had an 'escalating CM' eliminated during months 4-6. The CM and non-CM groups were compared using thrice weekly urine samples. SETTING: Out-patient buprenorphine maintenance for 6 months. PARTICIPANTS: All 75 of the 160 original study patients who completed month 3 of the clinical trial. INTERVENTION: The 'escalating CM' was eliminated for all 3 months and during months 5 and 6 the response requirement was also increased to two and then three consecutive drug-free urines in order to obtain a voucher. MEASUREMENTS: Urine toxicology for opiates and cocaine. FINDINGS: After eliminating the 'escalating CM', the CM group showed a decline in combined opioid and cocaine-free urines. This decline within the CM group was greater in those treated with DMI than placebo. CONCLUSIONS: Buprenorphine with DMI maintained drug abstinence after eliminating the 'escalating CM', but not after increasing the response requirement, suggesting the need for more intensive psychosocial interventions during CM. PMID- 12751986 TI - A longitudinal study of the effects of adolescent cannabis use on high school completion. AB - OBJECTIVE: To examine the extent to which weekly cannabis use during mid adolescence may increase the risk of early school-leaving. SETTING: A prospective study of a general population sample of adolescents studied from ages 15-21 years in Melbourne, Australia. METHOD: Computer-assisted self-completion questionnaires and telephone interviews conducted in six waves at ages 15-18 and again at age 21 in a sample of 1601 male and female school students. RESULTS: Weekly cannabis use, assessed prospectively, was associated with significantly increased risk of early school-leaving. This effect remained after adjustment for a range of prospectively assessed covariates including demographic characteristics, other substance use, psychiatric morbidity and antisocial behavior. There was suggestive evidence of an interaction between weekly cannabis use and age with the effects of weekly cannabis use on early school-leaving being strongest at the youngest ages and diminishing progressively with age. CONCLUSIONS: Early regular cannabis use (weekly use at age 15) is associated with increased risk of early school-leaving. These effects of regular cannabis use may diminish with increasing age and are likely to operate through the social context within which cannabis is used and obtained. PMID- 12751985 TI - Does retention matter? Treatment duration and improvement in drug use. AB - AIM: This study examines whether there is a minimum threshold, continuous or non linear relationship between the duration of addiction treatment and improvements in drug use. DESIGN: Longitudinal cohort study of 62 drug treatment units and 4005 clients in the US National Treatment Improvement Evaluation Study, fielded from 1993 to 1995. SUBJECTS: Baseline and 1-year follow-up interviews with clients in methadone maintenance, out-patient non-methadone, short-term residential and long-term residential treatment programs. MEASURES: Improvement in drug use is the difference between the client-reported peak frequency of drug use (in days per month) in the year prior to the baseline interview minus the peak frequency in the year after discharge. Primary drug, and overall use of the major illicit drugs (heroin, cocaine powder, crack cocaine, and marijuana) are considered separately. RESULTS: Controlling for multiple factors, treatment duration had a positive linear relationship with primary drug use improvement among methadone clients and an inverted-U-shaped relationship with overall and primary drug use improvements among out-patient and long-term residential clients. Improvement with longer duration is greatest for long-term residential clients. CONCLUSIONS: Contrary to previous arguments for a sharp retention threshold for onset of treatment effects, we find smooth curves relating treatment duration to drug use improvements in methadone maintenance, out-patient non-methadone and long-term residential modalities. These relationships are effectively linear for durations typically observed in single treatment episodes, but unusually long retention in out-patient non-methadone and long-term residential units appear steadily less predictive of improvement. PMID- 12751987 TI - Buprenorphine versus methadone maintenance treatment in an ambulant setting: a health-related quality of life assessment. AB - BACKGROUND: To compare the effects on quality of life (QOL) of oral methadone with sublingual buprenorphine. METHODS: We performed an open-label, non randomized, two-site (methadone-buprenorphine) study. During 6 months we assessed the quality of life status of 53 opioid-dependent patients admitted to a methadone or buprenorphine maintenance programme using the German version (Berlin Quality of Life Profile) of the Lancashire Quality of Life Profile. Physical symptoms were measured using the Opioid Withdrawal Scale. Five hundred and thirty urine screening tests were carried out randomly to detect additional consumption. RESULTS: Sixty-seven opioid-dependent subjects (38 on methadone and 29 on buprenorphine) were enrolled in the study, and 53 completed it (30 subjects treated with buprenorphine and 23 subjects with racemic methadone). The subjects were comparable on all baseline measures. At the first follow-up (week 8), the buprenorphine-maintained group showed significantly less additional consumption of opioids (P = 0.013) compared with the methadone group. Patients retained in the buprenorphine or methadone programme (week 24) showed no significant differences in all quality of life scores. At the end of the study period, the buprenorphine-maintained group showed significantly less additional consumption of opioids (P = 0.001) and cocaine (P = 0.018) compared with the methadone group. The outcome measures for withdrawal symptoms after 24 weeks of treatment with buprenorphine showed slight advantages in stomach cramps, fatigue or tiredness, feelings of coldness and heart pounding. CONCLUSIONS: These results suggest that buprenorphine treatment is as effective as methadone regarding effects on quality of life and withdrawal symptoms. Buprenorphine has the potential to reduce the harm caused by drug abuse. Further research is needed to determine if buprenorphine is more effective than methadone in particular subgroups of patients. PMID- 12751990 TI - Xenia Forsselliana. PMID- 12751992 TI - High-resolution CT in chronic pulmonary changes after mustard gas exposure. AB - PURPOSE: To identify the findings of high-resolution CT (HRCT) of the lung in patients with previous sulfur mustard gas exposure, and to correlate these findings with clinical and chest X-ray (CXR) results. MATERIAL AND METHODS: 50 consecutive patients were studied prospectively. The clinical data were recorded. Standard p.a. CXR and HRCT of the lung and spirometry were performed. The findings of CXR, HRCT and clinical and spirometry results were scored between 0 and 3 according to the severity of the findings. RESULTS: HRCT abnormality was detected in all 50 patients (100%), while CXR was abnormal in 40 patients (80%). The most common HRCT findings was airway abnormalities (bronchial wall thickening in 100% of cases). Other important findings were suggestive of interstitial lung disease (ILD) (80%), bronchiectasis (26%), and emphysema (24%). A statistically significant correlation was found between the severity of clinical presentation and that of the HCTR scores in patients with bronchiectasis, bronchitis and ILD (p< 0.05), but not with severity scores of HRCT in patients with emphysema. No significant correlation was found between severity scores of CXR findings. HRCT evidence of bronchial wall thickening and with a lower frequency ILD were present despite normal CXR in 20% of the patients. CONCLUSION: The results of this study suggest that bronchial wall thickening, ILD and emphysema are common chronic pulmonary sequelae of sulfur mustard injury. HRCT of the chest should be considered as the imaging modality of choice in chemical war injury. PMID- 12751993 TI - Lung perfusion scintigraphy prior to lung volume reduction surgery. AB - PURPOSE: The aim of this study was to evaluate whether lung perfusion scintigraphy (LPS) contributes to the preoperative classification of emphysema heterogeneity in patients undergoing LVRS (lung volume reduction surgery) compared to classification based only on computed tomography (CT). MATERIAL AND METHODS: Forty-five potential candidates for LVRS were examined with CT and LPS. The distribution of emphysema within the lungs was visually classified into three categories: markedly heterogeneous, intermediately heterogeneous, or homogeneous. The results of the two imaging techniques were compared to an objective, CT-based computerized classification of heterogeneity. RESULTS: Visual evaluation of all 90 lungs resulted in 50 correct classifications based on CT, in 40 based on LPS and in 68 correct classifications based on the combination of CT and LPS. The combination was superior to CT alone (p<0.01) in classification of emphysema heterogeneity. There was no significant difference between the evaluations based on either CT or LPS. CONCLUSION: The combined information from CT and LPS are superior in assessing emphysema heterogeneity prior to LVRS. PMID- 12751994 TI - Pulmonary nodule detection using chest CT images. AB - PURPOSE: Automated methods for the detection of pulmonary nodules and nodule volume calculation on CT are described. MATERIAL AND METHODS: Gray-level threshold methods were used to segment the thorax from the background and then the lung parenchyma from the thoracic wall and mediastinum. A deformable model was applied to segment the lung boundaries, and the segmentation results were compared with the thresholding method. The lesions that had high gray values were extracted from the segmented lung parenchyma. The selected lesions included nodules, blood vessels and partial volume effects. The discriminating features such as size, solid shape, average, standard deviation and correlation coefficient of selected lesions were used to distinguish true nodules from pseudolesions. With texture features of true nodules, the contour-following method, which tracks the segmented lung boundaries, was applied to detect juxtapleural nodules that were contiguous to the pleural surface. Volume and circularity calculations were performed for each identified nodule. The identified nodules were sorted in descending order of volume. These methods were applied to 827 image slices of 24 cases. RESULTS: Computer-aided diagnosis gave a nodule detection sensitivity of 96% and no false-positive findings. CONCLUSION: The computer-aided diagnosis scheme was useful for pulmonary nodule detection and gave characteristics of detected nodules. PMID- 12751995 TI - Interstitial lung disease in systemic sclerosis. AB - PURPOSE: To evaluate high-resolution CT (HRCT) parameters of inflammation and fibrosis in systemic sclerosis (SSc), for correlation with lung function, skin scores and exercise tolerance. MATERIAL AND METHODS: 45 SSc patients (40 women, 48.5+/-13.4 years), underwent thoracic HRCT, lung function assessment, and modified Rodnan skin scores. Exercise tolerance was also graded. HRCT were scored for extent of 4 HRCT patterns of interstitial lung disease (ILD): ground glass opacification (GGO), reticular, mixed and honeycomb pattern in each lobe. Total HRCT score, inflammation index (GGO and mixed score) and fibrosis index (reticular and honeycomb scores) were correlated with lung function and clinical parameters. RESULTS: ILD was present in 39/45 (86.7%) patients. Abnormal (<80% predicted) forced vital capacity (FVC), total lung capacity (TLC) and carbon monoxide diffusion factor (DLco) were detected in 30%, 22% and 46% of patients. Total HRCT score correlated with FVC (r=-0.43, p=0.008), FEV1 (forced expiratory volume) (r=-0.37, p=0.03), TLC (r=-0.47, p=0.003), and DLCO (r=-0.43, p=0.008); inflammatory index with DLCO (r=-0.43, p=0.008) and exercise tolerance (r=-0.39, p < 0.05); and fibrosis index with FVC (r=-0.31, p=0.05) and TLC (r=-0.38, p=0.02). Higher total HRCT score, and inflammation and fibrosis indices were found in patients with abnormal lung function. CONCLUSION: Qualitative HRCT is able to evaluate inflammation and fibrosis, showing important relationships with diffusion capacity and lung volume, respectively. PMID- 12751996 TI - Simultaneous voiding cystourethrography and voiding urosonography. AB - PURPOSE: To compare the diagnostic accuracy of contrast-enhanced voiding urosonography (VUS) and voiding cystourethrography (VCUG) during simultaneous performance of both examinations. MATERIAL AND METHODS: A total of 24 children, 16 girls and 8 boys, with a mean age of 3.5 years referred for reflux examination were recruited for the study. After transurethral bladder catheterization, radiographic contrast medium, followed directly by the US contrast medium, were administered. Fluoroscopic VCUG and VUS were carried out concurrently in the same patient. When 1 kidney was scanned by ultrasound, fluoroscopy was performed on the contralateral side. RESULTS: In 19 of the 47 kidney-ureter-units (KUU) vesicoureteral reflux (VUR) was detected. In 16 units the reflux was detected by both VCUG and VUS. In 3 KUUs the reflux was detected only at VCUG. All 3 cases were grade 1. Taking the VCUG as the reference standard, VUS had 84% sensitivity, 100% specificity, 100% and 90% positive and negative predictive values, respectively. CONCLUSION: A dependable comparison could be achieved by performing VCUG and VUS at the same time and under the same conditions. It reconfirmed that VUS is reliable in the exclusion or verification of reflux. PMID- 12751997 TI - Transvaginal power Doppler ultrasound in pelvic congestion. AB - PURPOSE: Pelvic congestion is diagnosed by transuterine venography, an invasive procedure requiring sedation and irradiation. Ultrasound may be an alternative but is hindered by slow flow within pelvic veins. In an attempt to counter this, we investigated the possible role of transvaginal power Doppler ultrasound. MATERIAL AND METHODS: 42 women with a clinical suspicion of pelvic congestion underwent transvaginal ultrasound. Adnexal veins were examined and a congestion score established. Planimetric measurements of adnexal vessels were obtained using power Doppler ultrasound, and uterine and ovarian morphology noted. All women then underwent transuterine venography and agreement with the ultrasound congestion score and morphologic features was determined. RESULTS: There was a trend towards weak positive correlation between ultrasound and venography congestion scores (r = 0.29, p = 0.06). However, agreement between scores was poor on an individual basis (95% limits of agreement, -3.9 to +2.7). Planimetric power Doppler assessments of adnexal vascularity were unrelated to venographic congestion. Instead, there was correlation between the number and diameter of ovarian follicles and venographic congestion: women with congestion tended to have significantly more (0.04) and smaller follicles (p = 0.001). CONCLUSION: There was poor individual agreement between ultrasound and venographic estimates of congestion. However, there was a direct relationship between venographic congestion score and ovarian morphology. PMID- 12751998 TI - New intravascular contrast agent applied to dynamic contrast enhanced MR imaging of human breast cancer. AB - PURPOSE: To evaluate the feasibility of using dynamic contrast-enhanced MR imaging with a new intravascular contrast agent in grading human breast cancer. MATERIAL AND METHODS: 23 patients with 27 breast tumors (21 carcinomas and 6 fibroadenomas) were examined with dynamic MR imaging after administration of Clariscan, an iron oxide nanoparticle with large T1 relaxivity and a long plasma half life. A 3D T1-weighted gradient echo sequence with an acquisition time of 60 s was repeated at regular intervals of 3-5 min before and up to 1 h after injection of 2 mg/kg b.w. of Clariscan. The endothelial transfer constant, Kps, which reflects overall vascular permeability, and the fractional plasma volume, fPV, were estimated from time-intensity curves acquired from three separate regions of interest (ROIs): whole tumor, a permeability hot spot, and a blood volume hot spot. Kps and fPV were compared to the results of histologic tumor grading (Scarff-Bloom-Richardson, SBR) and microvascular density, MVD. RESULTS: A statistically significant correlation between the MR-derived Kps parameters and the SBR score was obtained for the whole tumor ROI (R = 0.70), and for the permeability hot spot ROIs (R = 0.67). A correlation between fPV and SBR was detected for the blood volume hot spot ROIs (R = 0.48). There was no statistically significant correlation between Kps or fPV with MVD. CONCLUSION: The results support the hypothesis that dynamic MR with the intravascular contrast agent Clariscan may be used for non-invasive tumor grading. PMID- 12751999 TI - Early assessment of neoadjuvant chemotherapy by FEC-courses of locally advanced breast cancer using 99mTc-MIBI. AB - PURPOSE: Response assessment at neoadjuvant (preoperative) chemotherapy of locally advanced breast cancer using clinical examination and mammography is insensitive. Mammoscintigraphy with 99mTc-MIBI was studied for the prediction of response at such therapy before finishing the chemotherapy cycles. MATERIAL AND METHODS: Chemotherapy was given as repeated courses of 5-fluorouracil, epirubicin and cyclophosphamide (FEC). In 1 patient group (n = 23), the tumor uptake relative to surrounding breast tissue and lung tissue at SPECT examination after finishing neoadjuvant chemotherapy was compared with the examination made before chemotherapy. In another group (n = 30), a similar comparison after the first therapy cycle (mean 19 days) with a baseline examination was made. Histologic examination of the resected tumors was made. RESULTS: After finishing chemotherapy, there was a strong reduction of the relative tumor activity, while there was no correlation with therapy effect as assessed by histology. After one therapy course, there was no significant reduction of the relative tumor uptake. CONCLUSION: Scintigraphy with 99mTc-MIBI demonstrated the response after finished neoadjuvant chemotherapy of breast cancer using FEC-courses. It cannot be used to predict a therapy response after one therapy course. PMID- 12752000 TI - Complementary use of scintimammography with 99m-Tc-MIBI to triple diagnostic procedure in palpable and non-palpable breast lesions. AB - PURPOSE: The aim of this study was to determine the clinical value of scintimammography with 99m-Tc-MIBI (Sc) as a complementary method to the triple diagnostic procedure in the diagnosis of breast lesions. MATERIAL AND METHODS: Ninety-six consecutive patients with 65 palpable and 54 non-palpable breast lesions were included in a prospective study. All lesions were evaluated by clinical examination, mammography and fine-needle-aspiration cytology (FNAC), called triple diagnostic procedure (TD). Prone planar scintimammography with 99m Tc-MIBI was performed in all patients. Five groups were defined for diagnosis: 1 = normal; 2 = benign; 3 = probably benign; 4 = highly suspect of malignancy; and 5 = malignant. In the calculations, groups 1-3 were considered benign, and 4-5 malignant. All lesions were excised and examined histologically. The additional value of Sc + TD procedure was studied separately for palpable and non-palpable lesions. RESULTS: Histologically, 83 malignant and 36 benign lesions were found in the 119 breast lesions. Sensitivity for malignancy in palpable lesions of TD alone and of the combination TD + Sc were 95.6% and 100%, respectively. Sensitivity for malignancy in non-palpable lesions of TD and TD + Sc was 89.1% and 97.2%, respectively. CONCLUSION: Adding scintimammography to the triple diagnostic procedure increased the sensitivity for the detection of both palpable and non-palpable breast cancers, but decreased the specificity. PMID- 12752001 TI - Elective placement of covered stents in native coronary arteries. AB - PURPOSE: To study the feasibility of placing a polytetrafluoroethylene (PTFE) covered stent graft into native coronary arteries and assess the complications and the restenosis rate. MATERIAL AND METHODS: Fifty consecutive patients with stable angina pectoris were included and the stent graft was placed into native coronary arteries. Clinical and angiographic follow-up were performed after 6 months. RESULTS: The stent grafts were successfully placed in all patients. The mean reference diameter was 3.3 +/- 0.6 mm. During follow-up the stent grafts occluded in patients after 1, 2 and 2.5 months and one more was occluded at 6 months. Three patients experienced myocardial infarction, 2 Q wave and one non-Q wave. After 6 months 42 (84%) patients had angina NYHA class 0 or 1. Target vessel revascularization was done in 11 cases for restenosis in the graft (n = 4), outside the graft (n = 3) and both (n = 4), giving a restenosis rate of 24%. The total major adverse coronary events at 6 months was 24%. CONCLUSION: The stent graft was deployed with a high success rate. The restenosis rate was not higher than expected for bare stents. However, this study showed that subacute occlusion may occur more frequently and we therefore recommend that ticlopidine or clopidogrel treatment should be prolonged to at least 3 months. PMID- 12752002 TI - Value of 3D-volume rendering in the assessment of coronary arteries with retrospectively ECG-gated multislice spiral CT. AB - PURPOSE: To assess the diagnostic value and measurement precision of 3D volume rendering technique (3D-VRT) from retrospectively ECG-gated multislice spiral CT (MSCT) data sets for imaging of the coronary arteries. MATERIAL AND METHODS: In 35 patients, retrospectively ECG-gated MSCT of the heart using a four detector row MSCT scanner with a standardized examination protocol was performed as well as quantitative X-ray coronary angiography (QCA). The MSCT data was assessed on segmental basis using 3D-VRT exclusively. The coronary artery diameters were measured at the origin of each main coronary branch and 1 cm, 3 cm and 5 cm distally. The minimum, maximum and mean diameters were determined from MSCT angiography and compared to QCA. RESULTS: A total of 353 of 525 (67.2%) coronary artery segments were assessable by MSCT angiography. The proximal segments were more often assessable when compared to the distal segments. Stenoses were detected with a sensitivity of 82.6% and a specificity of 92.8%. According to the Bland-Altman method the mean differences between QCA and MSCT ranged from -0.55 to 1.07 mm with limits of agreement from -2.2 mm to -2.7 mm. CONCLUSION: When compared to QCA, the ability of 3D-VRT to quantitatively assess coronary artery diameters and coronary artery stenoses is insufficient for clinical purposes. PMID- 12752003 TI - CT pulmonary angiography and suspected acute pulmonary embolism. AB - PURPOSE: To evaluate the use and quality of CT pulmonary angiography in our department, and to relate the findings to clinical parameters and diagnoses. MATERIAL AND METHODS: A retrospective study of 324 consecutive patients referred to CT pulmonary angiography with clinically suspected pulmonary embolism (PE). From the medical records we registered clinical parameters, blood gases, D-dimer, risk factors and the results of other relevant imaging studies. RESULTS: 55 patients (17%) had PE detected on CT. 39 had bilateral PE, and 8 patients had isolated peripheral PE. 87% of the examinations showing PE had satisfactory filling of contrast material including the segmental pulmonary arteries, and 60% of the subsegmental arteries. D-dimer test was performed in 209 patients, 85% were positive. A negative D-dimer ruled out PE detected at CT. Dyspnea and concurrent symptoms or detection of deep vein thrombosis (DVT), contraceptive pills and former venous thromboembolism (VTE) were associated with PE. The presence of only one clinical parameter indicated a negative PE diagnosis (p < 0.017), whereas two or more suggested a positive PE diagnosis (p < 0.002). CT also detected various ancillary findings such as consolidation, pleural effusion, nodule or tumor in nearly half of the patients; however, there was no association with the PE diagnosis. CONCLUSION: The quality of CT pulmonary angiography was satisfactory as a first-line imaging of PE. CT also showed additional pathology of importance in the chest. Our study confirmed that a negative D-dimer ruled out clinically suspected VTE. PMID- 12752004 TI - MR angiography for the evaluation of non-systemic vasculitic neuropathy. AB - Peripheral neuropathy due to vasculitis without any complications of vasculitis in other organs was first reported in 1987. This condition was termed non systemic vasculitic neuropathy (NSVN). Although vasculitis is believed to develop in small arteries and arterioles in this disease, the level of vascular involvement has not been fully established. We present a case of NSVN followed up by MR angiography, which was thought to be useful to assess the level as well as the state of vascular lesions in this condition. PMID- 12752005 TI - Modified carpal box technique in the diagnosis of suspected scaphoid fractures. AB - PURPOSE: To establish and test the clinical efficacy of a new diagnostic algorithm with the extensive utilization of modified carpal box radiography (mX CB) in the detection of scaphoid fractures. MATERIAL AND METHODS: Initial and early follow-up radiographic evaluation of 146 suspected scaphoid fractures were carried out by mX-CB. Patients with unconfirmed diagnosis were referred to CT. Patients were followed for 1 year after injury. Sensitivity, specificity and interobserver agreement of reading mX-CB images were determined statistically. RESULTS: No non-union or avascular necrosis was seen at 1 year after the injury. 90% of the fractures were diagnosed by mX-CB, only 6.8% of the patients needed referral to CT. Sensitivity of mX-CB at initial presentation was 81.6%. Interobserver agreement was very high among evaluators of mX-CB images. CONCLUSION: Extensive utilization of mX-CB as primary and early follow-up investigation resulted in high initial diagnostic accuracy and low referral rate to a more expensive diagnostic modality. PMID- 12752006 TI - MR imaging of uncommon recurrence of fibrolipomatous hamartoma of the ulnar nerve. AB - We present the MR and histopathologic findings of fibrolipomatous hamartoma (FLH) of the ulnar nerve in a 54-year-old woman, a lipomatous process that rarely affects the ulnar nerve. The case illustrated is further unusual as a local soft tissue recurrent mass developed over a remarkably long course of the disease. PMID- 12752007 TI - Imaging of esophageal tuberculosis: a review of 23 cases. AB - PURPOSE: To evaluate the various radiological abnormalities in patients with proven esophageal tuberculosis. MATERIAL AND METHODS: The case records of 23 patients with proven esophageal tuberculosis were evaluated retrospectively for various radiological abnormalities. Twenty-two patients had secondary involvement of esophagus in the form of direct extension of mediastinal and pulmonary tuberculosis or spinal tuberculosis. Only 1 patient had primary involvement of the esophagus with no evidence of disease elsewhere. The diagnosis was confirmed by endoscopic and CT-guided biopsy/aspiration cytology in 7 and 6 cases, respectively. Diagnosis was made on the basis of surgical biopsy of lymph node and autopsy in 1 patient each. In the remaining 8 patients the diagnosis was based on radiological and endoscopic findings and the response to antituberculous treatment. RESULTS: Chest radiography (CXR) was abnormal in 65% patients. While the findings were non-conclusive for esophageal tuberculosis, characteristic lesions of tuberculosis in lungs or spine were suggestive of tuberculous etiology. In 15 patients, CT of the chest confirmed the corresponding CXR findings and also showed additional findings of mediastinal lymphadenopathy when CXR was normal. Fourteen patients showed mediastinal lymphadenopathy on CT of the chest. In all these patients, more than one group of lymph nodes was involved. The characteristic hypodense center of lymph nodes suggestive of tuberculosis was seen in 12 patients. Radiological abnormalities seen in barium swallow examination were extrinsic compression, traction diverticula, strictures, sinus/fistulous tracts, kinking and pseudotumor mass of esophagus in decreasing order of frequency. The middle third of the esophagus was found to be the most frequent site of involvement. PMID- 12752008 TI - Morphologic and histopathologic changes in the bowel after super-selective transcatheter embolization for focal lower gastrointestinal hemorrhage. AB - PURPOSE: To evaluate the safety of superselective arterial embolization therapy in the lower gastrointestinal tract. The sequelae on normal enteric tissue in lower gastrointestinal arterial embolization were retrospectively reviewed. MATERIAL AND METHODS: To control hemorrhage and tumor blood supply, 14 patients were treated by superselective transcatheter embolization at different levels of the colonic and small intestine vessels via the superior and inferior mesenteric arteries using microcoils and/or gelatin sponge. Normal enteric tissues in the embolized areas were analyzed for the occurrence of ischemic changes by clinical follow-up, colonoscopy, and surgery. RESULTS: Normal bowel function was preserved in 13 patients. In 1 patient treated with numerous gelatin sponge particles delivered from the proximal arcade of the superior mesenteric artery, significant muscular fibrosis occurred. CONCLUSION: Superselective arterial embolization for lower gastrointestinal hemorrhage can be safely performed by minimizing the amount of embolic materials and delivering them as distally as possible. PMID- 12752009 TI - A modified sonographic technique for assessment of gastric emptying of liquid. AB - PURPOSE: Ultrasound scanning is replacing scintigraphy in studies of gastric emptying of liquid, but both have considerable day-to-day variability. This study describes a modified ultrasound technique for assessing gastric emptying of liquid, and evaluates the inter- and intraindividual variation in emptying time. MATERIAL AND METHODS: On different days, each of 12 healthy volunteers had meals of 350 ml broth. The antral area was measured at sonography 5 times before the meal as a baseline, and every 1-4 min after the meal. The time until the antral area had decreased to 150% of baseline (T150) was determined and used as surrogate expression of gastric emptying time. RESULTS: The mean T150 for a broth meal was 12.6 min (range 5-21) and 13.5 min (6-23) (first and second meal, respectively). The standard deviation of the differences between the 12 pairs of repeated measurements was 3.1 min and the coefficient of variation was 24%. CONCLUSION: Ultrasound monitoring of antral size after a liquid meal is a well suited method for assessing gastric emptying of liquid. PMID- 12752010 TI - Delayed diffusion-weighted MR abnormality in a patient with an extensive acute cerebral hypoxic injury. AB - Diffusion-weighted (DW) MR imaging usually identifies acute cerebral infarction injury in symptomatic patients. We report a patient with severe hypoxic brain injury following suicide attempt by hanging, but with normal DW MR imaging 5-6 h after the event. Follow-up DW MR imaging 3 days after the event, and subsequent autopsy, revealed extensive cerebral anoxic injury. PMID- 12752011 TI - Altered response in renal blood flow and oxygen tension to contrast media in diabetic rats. AB - PURPOSE: To investigate the effect of the contrast medium (CM) iopromide on renal microcirculation and oxygen tension in non-diabetic control and streptozotocin diabetic Wistar Furth rats. MATERIALS AND METHODS: Oxygen tension was measured with Clark-type microelectrodes and blood flow with laser-Doppler flow probes. In order to differentiate between an acutely increased blood glucose concentration and a long-term diabetic state, some of the non-diabetic control rats were intravenously infused with glucose. RESULTS: CM decreased the medullary oxygen tension in control (non-diabetic normoglycemic) rats ( approximately 35%) but not in diabetic rats. Medullary blood flow in control rats increased after CM administration and remained elevated, while it was unchanged in the diabetic rats. In response to CM, glucose-infused control rats responded similarly to control animals in medullary oxygen tension, but similarly to diabetic rats in medullary blood flow. Contrary to in control rats, medullary oxygen tension was unchanged in diabetic animals after CM administration. CONCLUSION: Streptozotocin diabetic rats have an altered response to intravenous injection of the CM iopromide compared to non-diabetic rats. The unaltered medullary oxygen tension, seen in the diabetic group after injection of CM, suggests that non-hemodynamic mechanisms are responsible for the increased frequency of renal failure commonly seen among diabetic patients. PMID- 12752012 TI - Is long-term use of a calcium channel blocker associated with an exacerbated response of an insulinoma during selective intraarterial calcium stimulation? PMID- 12752013 TI - Remodeling of the temporal bone after involvement with Langerhans' cell histiocytosis. PMID- 12752014 TI - Shared decision making: a model for the physician-patient relationship in the 21st century? PMID- 12752015 TI - Shared decision making in psychiatry. AB - OBJECTIVE: In other medical fields, 'shared decision making' (a strategy for including patients in therapeutic decision processes) has been studied for several years now. This survey presents the potential of shared decision making for psychiatry. METHOD: Literature search. RESULTS: Research in other medical fields shows that shared decision making positively influences patients' satisfaction as well as health outcomes. The application of shared decision making to psychiatry is still in the initial stage. CONCLUSION: Evidence that shared decision making can be successfully practiced in psychiatry would contribute to an improved inclusion of psychiatric patients in therapeutic decisions and thereby help to implement basic rights of a group of patients who have not sufficiently benefited from consumer empowerment in other medical fields. PMID- 12752016 TI - User involvement in the planning and delivery of mental health services: a cross sectional survey of service users and providers. AB - OBJECTIVE: To identify methods for involving service users in the planning and delivery of psychiatric services and factors which may assist and impede this process. METHOD: A cross-sectional postal survey of user groups and providers of psychiatric services throughout Greater London (UK). RESULTS: Seventeen (94%) service providers and 29 (48%) user groups responded to the survey. Service providers employed a wide variety of different methods for involving users but none met national standards for user involvement (UI). Service providers stated that the main obstacle to UI was that users who took part were not representative of local patients. User groups highlighted staff resistance as a major obstacle and 80% stated that they were not satisfied with current arrangements for UI. CONCLUSION: While users and providers of mental health services were able to identify changes resulting from UI the responsiveness of staff and the representativeness of service users may be impeding this process. PMID- 12752017 TI - Integrated care in schizophrenia: a 2-year randomized controlled study of two community-based treatment programs. AB - OBJECTIVE: To evaluate the efficacy of two community-based programs that combined antipsychotic medication, family interventions and social skills training. METHOD: A randomized controlled trial with 2 years follow-up. The study included 84 patients with schizophrenic disorders, continuously managed in terms of care and treatment, and regularly assessed. Analysis was by intention-to-treat. RESULTS: Between-program comparisons showed significantly improved social function and consumer satisfaction in favour of the program 'Integrated Care' (IC) at the 2-year follow-up. The main clinically important differences between the two treatment programs studied were the procedures for shared decision making and patient empowerment in IC. CONCLUSION: The implementation of IC in clinical practice can improve social recovery and increase consumer satisfaction for patients with schizophrenic disorders. We identified specific procedures that might be added to improve the effectiveness of any program for severely mental ill people. PMID- 12752018 TI - Attitudes toward psychiatry among students entering medical school. AB - OBJECTIVE: To survey the attitudes of Australian medical students to determine their views about the relative attractiveness of psychiatry as a career compared with other specialties, and against findings from a North American study. METHOD: We surveyed 655 first-year medical students attending six Australian Universities. RESULTS: Responses indicated that Australian medical students view psychiatry as distinctly less 'attractive' than other career options, as reported in the North American sample. In comparison with other disciplines, psychiatry was regarded as more interesting and intellectually challenging, but also as lacking a scientific foundation, not being enjoyable and failing to draw on training experiences. CONCLUSION: Our findings suggest that psychiatry has an image problem that is widespread, reflecting community perceptions and the specialist interests of medical students on recruitment. If psychiatry is to improve its 'attractiveness' as a career option, identified image problems need to be corrected and medical student selection processes re-considered. PMID- 12752019 TI - Suicide attempts among patients with psychosis in a Chinese rural community. AB - OBJECTIVE: To explore the characteristics of psychotic patients with suicide attempts in a Chinese rural community. METHOD: An epidemiological investigation of psychotic patients with suicide attempts among 123,572 population (over 15 years of age) was conducted in Xinjin County, Chengdu. RESULTS: The rate of suicide attempts was found to be 8.17% among all the psychotic patients (906 cases). Patients with affective psychosis showed a significantly higher rate of suicidal attempts (15.3%) than those with schizophrenia (7.5%) (P < 0.005). Suicide attempts were significantly associated with depression and hopelessness in both schizophrenia and affective psychosis (P < 0.001). Patients with suicide attempts were younger and had an earlier age of onset than those without suicide attempts (P < 0.05). Patients with schizophrenia and affective disorders were the major patients with suicide attempts. CONCLUSION: The rate of suicide attempts in psychotic patients may be largely influenced by the illness itself. Community based services should be necessary for these patients. PMID- 12752020 TI - Comorbid personality disorder predicts suicide after major depression: a 10-year follow-up. AB - OBJECTIVE: To identify psychopathological predictors for suicide in a population of major depressed Diagnostic Statistical Manual-III (DSM-III) in-patients. METHOD: A total of 210 previous participants in multicentre antidepressant drug trials, carried out in a randomized double-blind design, were followed prospectively through a maximum of 10 years. Patients with a drug or alcohol abuse were excluded. The association between suicide and the pretreatment psychopathological profile was analysed using survival statistics. RESULTS: The suicide rate for non-melancholic depressed patients was significantly higher than for melancholic depressed patients. Comorbid personality disorder was independently associated with an increased suicide rate [relative hazard 3.41(CI: 1.15-10.10)]. CONCLUSION: The study indicates that the non-melancholic aspect of depression, and especially comorbid personality disorder, is associated with an increased suicidal vulnerability. PMID- 12752021 TI - Does co-morbid personality disorder increase the risk of suicidal behaviour in psychosis? AB - OBJECTIVE: To examine the association between co-morbid personality disorder (PD) and suicidal behaviour over a 2-year period in a sample of patients with psychosis. METHOD: A total of 670 patients with established psychotic illness were interviewed using a battery of instruments including a screen for co-morbid PD. The prevalence of attempted and completed suicide was measured over the next 2 years using multiple data sources. Logistic regression was used to examine whether those with co-morbid PD were at greater risk of suicidal behaviour compared with others. RESULTS: One hundred and eighty six patients (28%) were rated as having a co-morbid PD. After adjusting for all covariates, patients with co-morbid PD were significantly more likely to attempt or complete suicide over the 2-year period (adjusted odds ratio: 1.87; 95% CI: 1.02-3.42). CONCLUSION: Co morbid PD is independently associated with an increased risk of suicidal behaviour in psychosis. Early assessment of personality status should be part of the routine assessment of all psychiatric patients. PMID- 12752022 TI - Group therapy for somatization disorders in primary care: maintenance of treatment goals of short cognitive-behavioural treatment one-and-a-half-year follow-up. AB - OBJECTIVE: The objective of this study was to evaluate the maintenance of treatment goals of a short cognitive-behavioural group treatment programme for the management of somatization disorders in primary care. METHOD: In a previous controlled 6-month follow-up study, patients with somatization disorders (n=32) improved with respect to illness and somatic preoccupation, hypochondriasis, and medication usage. In the present report the same group of patients were also investigated one-and-a-half year after initial treatment. RESULTS: The long-term follow-up manifested maintained improvement with respect to hypochondriasis. There was additional reduction of anxiety and psychosocial preoccupation, whereas somatization and depression-anxiety scores improved progressively. CONCLUSION: A short cognitive-behavioural group treatment of psychosomatic patients can be useful in primary care and may manifest maintained or progressive beneficial outcome. PMID- 12752023 TI - A follow-up study of juvenile obsessive-compulsive disorder from India. AB - OBJECTIVE: To study the long-term course and outcome of juvenile obsessive compulsive disorder (OCD). METHOD: Two to 9-year follow-up of largely self referred, drug-naive subjects (n = 58) by employing catch-up longitudinal design. RESULTS: The mean follow-up period was 5 years. Nearly three-fourth of the sample was adequately treated with medications. Only 21% of the subjects had clinical OCD at follow-up and 48% were in true remission (no OCD and not on treatment). Earlier age-at-onset was associated with better course and outcome. CONCLUSION: Juvenile OCD has favorable outcome. Our findings are applicable to psychiatric hospital settings in India and perhaps to the general psychiatric settings in the Western countries. Whether the better outcome in this sample is the result of differing clinical characteristics or because of true cross-cultural variation in the course needs further exploration. It is speculated that early onset OCD could be a subtype of juvenile OCD with better outcome. PMID- 12752024 TI - A case study of an extremely violent serial rapist with borderline personality disorder in remission. AB - OBJECTIVE: To provide evidence that some borderline patients with poor prognosis can obtain complete remission. METHOD: Case of a violent rapist with borderline personality disorder in remission is reported. An attempt was made to reveal the mechanism of recovery in this forensic patient who initially was assessed as psychotherapeutically untreatable. RESULTS: Psychotherapeutic effects and favourable psychosocial factors such as successful college graduation, increased social-emotional capacities and valuable new relationships did also play a very important role in this remission process. CONCLUSION: Controlled studies are necessary to elucidate the mechanism of recovery among criminal patients with borderline personality disorder in order to provide adequate treatment programmes and to construct more reliable prediction methods for prognosis and reoffending. PMID- 12752026 TI - Attention-deficit hyperactivity disorder in a life perspective. PMID- 12752027 TI - Searching for schizophrenia in ancient Greek and Roman literature: a systematic review. AB - OBJECTIVE: The aim of this study was to systematically examine ancient Roman and Greek texts to identify descriptions of schizophrenia and related disorders. METHOD: Material from Greek and Roman literature dating from the 5th Century BC to the beginning of the 2nd Century AD was systematically reviewed for symptoms of mental illness. DSM IV criteria were applied in order to identify material related to schizophrenia and related disorders. RESULTS: The general public had an awareness of psychotic disorders, because the symptoms were described in works of fiction and in historical accounts of malingering. There were isolated instances of text related to psychotic symptoms in the residents of ancient Rome and Greece, but no written material describing a condition that would meet modern diagnostic criteria for schizophrenia. CONCLUSION: In contrast to many other psychiatric disorders that are represented in ancient Greek and Roman literature, there were no descriptions of individuals with schizophrenia in the material assessed in this review. PMID- 12752028 TI - Prenatal exposure to influenza as a risk factor for adult schizophrenia. AB - OBJECTIVE: Several, but not all epidemiological studies, have demonstrated a positive correlation between exposure to the virus during the second trimester of pregnancy and an increased risk to the infants for subsequently developing schizophrenia. The present study is the first be designed in France to examine the risk of gestational exposure to the influenza virus and subsequent development of schizophrenia. METHOD: A total of 974 adults with schizophrenia born between 1949 and 1981 were compared for risk of exposure to influenza with their non-schizophrenic siblings and with matched control patients. RESULTS: Significantly more schizophrenic subjects than controls (both groups) had been exposed to the influenza virus during the fifth month of pregnancy (OR=2.24, CI: 1.49-3.35, and OR=1.61, CI: 1.04-2.49). CONCLUSION: These results suggest that influenza infection during pregnancy is a neurodevelopmental risk factor for schizophrenia in adult life. PMID- 12752029 TI - Aging and outcome in schizophrenia. AB - OBJECTIVE: Controversy exists about long-term outcome of schizophrenia, but few studies have compared older out-patients to normal subjects. We sought to examine the relationship of age to clinical features, psychopathology, movement abnormalities, quality of well-being, and everyday functioning in schizophrenia out-patients and normal comparison subjects, and to further characterize these outcomes in elderly schizophrenia out-patients. METHOD: A total of 290 out patients and 144 comparison subjects, aged 40-85 years, underwent comprehensive assessments. RESULTS: Among patients, aging was associated with decreased psychopathology, even after controlling for duration of illness. There was no accelerated aging-related decline on any measure in the patients. Yet, elderly patients were more impaired than comparison subjects on various measures. CONCLUSION: The course of schizophrenia in late life appears stable, but most elderly patients remain symptomatic and impaired. Our findings dispute notions of either progressive deterioration or marked improvement in aging schizophrenia out patients. PMID- 12752030 TI - Long-term effectiveness of risperidone and olanzapine in resistant or intolerant schizophrenic patients. A mirror study. AB - OBJECTIVE: To evaluate the long-term effectiveness of the second generation antipsychotics, risperidone and olanzapine on hospitalization, clinical response, and adherence to therapy after switching from conventional antipsychotics or clozapine in a naturalistic setting of schizophrenic patients. METHOD: Data were registered for periods of identical duration before and after switching. RESULTS: Fifty-seven patients were included. Mean study period was 3.1 +/- 0.2 years. Mean number of in-patient days after switching declined by 78% of the level before switching (P=0.0002). There was no significant differences in ratio responders/non-responders between the subgroups. The number of patients with extrapyramidal symptoms (EPS) was significantly reduced after switching. However, intolerable weight gain led to a lack of adherence to therapy in 18% of the olanzapine-treated patients. CONCLUSION: Long-term treatment with the second generation antipsychotics, risperidone and olanzapine significantly improved the clinical outcome. However, weight gain may be a significant reason for discontinuation of treatment and reducing the long-term adherence to therapy. PMID- 12752031 TI - A longitudinal analysis of the impact of social and clinical characteristics on the costs of schizophrenia treatment. AB - OBJECTIVE: The aim of the study was the longitudinal analysis of the influence of social and clinical factors on the mid-term costs of schizophrenia treatment. METHOD: Treatment costs as well as clinical and social characteristics of 307 patients with the ICD-10 diagnosis of schizophrenia were assessed at five follow ups over 2.5 years. Between and within effects of clinical and social characteristics on treatment costs were estimated by error component regression models. RESULTS: Effects caused by differences between individuals were found for age, partnership, in-patient history, objective and subjective role functioning, life-events and psychotic relapse. Effects of idiosyncratic transitory changes of social and clinical characteristics were found for symptoms, psychotic relapse, and for social role-functioning. CONCLUSION: Treatment costs can be reduced to a limited extent not only by the prevention of psychotic symptoms and relapse but also by the improvement of role-functioning capacities. PMID- 12752032 TI - Cost-effectiveness of the HIT programme in patients with schizophrenia and persistent auditory hallucinations. AB - OBJECTIVE: To examine the cost-effectiveness of Hallucination focused Integrative Treatment (HIT) in patients with schizophrenia and a history of persistent auditory hallucinations. METHOD: Costs, in and outside the health care sector, and outcomes were registered prospectively during a period of 18 months for patients who received the HIT programme and for patients in the care as usual (CAU) condition. The Positive and Negative Syndrome Scale (PANSS) was used as main outcome measure in the cost-effectiveness analysis. Bootstrap analyses provided additional information on the skewly distributed costs. RESULTS: Mean costs per patient in the HIT group (18,237 dollars) were lower than the mean costs per patient in the CAU group (21,436 dollars). Results of the PANSS were slightly in favour of the HIT group. CONCLUSION: There appears to be no significant cost-effectiveness advantage of the HIT programme over CAU. Additional analyses indicated that future application of the HIT programme will, in most cases, lead to a reduction of (non) medical costs. PMID- 12752033 TI - Self-reported psychotic experiences in the general population: a valid screening tool for DSM-III-R psychotic disorders? AB - OBJECTIVE: To examine the diagnostic value of self-reported psychotic-like experiences for DSM-III-R psychotic disorders. METHOD: A general population sample of 7076 subjects aged 18-64 years was interviewed with the Composite International Diagnostic Interview (CIDI) and, if there was evidence of psychotic experiences, the Structured Clinical Interview for DSM-III-R. RESULTS: The probability of having a psychotic disorder increased in a dose-response fashion with the level of self-reported psychotic experiences, but individual CIDI psychotic experience ratings had relatively low post-test probabilities (PPs) (range: 5.1-26.5%). However, limiting the sample to individuals who had been in contact with mental health services substantially improved PPs (range: 13.3 43.1%). CONCLUSION: Screening for psychosis in the population carries a high risk of stigmatization in false-positive cases and violation of the right 'not to know' in true-positive cases. However, in mental health care users, self-reported psychotic experiences may be a useful screening tool in individuals who have already developed help-seeking. PMID- 12752034 TI - Dimensional vs. categorical diagnosis in psychosis. AB - OBJECTIVE: To compare dimensional measures vs. categorical diagnosis of psychopathology in their prediction of disability and outcome in psychotic illness. METHOD: A community study of 980 subjects with psychotic illness was included in the Australian National Survey of Mental Health and Wellbeing. The study instrument [including the Schedules for Clinical Assessment in Neuropsychiatry (SCAN)] yielded symptom data along with Operational Criteria for Psychotic Illness (OPCRIT) diagnoses by several diagnostic schemata. Factor analysis of symptoms yielded five dimensions of psychopathology (positive symptoms, negative symptoms, dysphoria, mania and substance use). Variance in service demand and disability explained by the diagnostic schemata was compared with the variance explained by the dimensions of psychopathology. RESULTS: Dimensional measures of psychopathology explained more of the variance in service demand, dysfunctional behaviour, social adaptation and global occupation and function. Only use of support services and illness course were better predicted by categorical diagnosis. Dimensional scores explained significant extra variance when added to categorical diagnoses. CONCLUSION: Dimensional measures of psychopathology explain more variance in behaviour, disability and outcome than does categorical diagnosis in functional psychosis. Dimensions provided significant extra information not provided by diagnosis and would be a more useful basis for clinical management. PMID- 12752035 TI - A psychopathological study into the relationship between attention deficit hyperactivity disorder in adult patients and recurrent brief depression. AB - OBJECTIVE: Clinical and epidemiological observations and neurobiological data suggest that there might be an inherent link between attention deficit hyperactivity disorder (ADHD) and recurrent brief depression (RBD). In this psychopathological study, we investigated the comorbidity between these two conditions. METHOD: Using an index patient approach 40 adult out-patients fulfilling the criteria for ADHD were investigated for lifetime history of RBD and another 40 out-patients with the primary diagnosis of RBD were investigated for a lifetime history of ADHD. RESULTS: We found a high prevalence of RBD in patients with ADHD (70%) while the prevalence of ADHD in the index sample with RBD was smaller (about 40%). CONCLUSION: In terms of comorbidity ADHD was the second commonest psychiatric disorder in patients with RBD next to other affective disorders. The psychopathological pattern of lifetime comorbidity might be of clinical relevance in terms of medical treatment. PMID- 12752036 TI - Prediction of transition from cognitive impairment to senile dementia: a prospective, longitudinal study. AB - OBJECTIVE: The purpose of this investigation was to replicate the statistical approach used in a previous investigation (Toronto study) within a French population to determine the best predictive model for Alzheimer's disease (AD). METHOD: Data from neuropsychological tests from two prospective studies were entered into a regression model. RESULTS: Replication of the statistical approach in the Montpellier sample produced a three-test model with a specificity of 99% and sensitivity of 73%. This model consisted of a delayed auditory verbal recall test, a construction test, a category fluency test and provides probability estimates for the transition to dementia in individual cases. CONCLUSION: The models derived from these two longitudinal studies provide an empirical basis for the selection of tests for the definition of mild cognitive impairment of the Alzheimer type (MCI-A). The small set of tests derived are suitable for use in general practice. PMID- 12752037 TI - Olanzapine in the treatment-resistant, combat-related PTSD--a series of case reports. AB - OBJECTIVE: Nightmares and insomnia in combat-related post-traumatic stress disorder (PTSD) might be resistant to treatment with selective serotonin reuptake inhibitors (SSRIs) and benzodiazepines. METHOD: We describe five case reports of patients suffering from long-lasting and intractable nightmares and insomnia. They were given different psychotropic agents in past few years, with no improvement in their sleep disturbance. Olanzapine was added to the current treatment regimen. RESULTS: Both nightmares and insomnia improved rapidly after olanzapine institution in all of five patients. No adverse events of olanzapine were reported. CONCLUSION: Olanzapine augmentation might be useful in alleviating treatment-resistant nightmares and insomnia in patients with combat-related PTSD. PMID- 12752045 TI - Recent advances in the treatment of exudative age-related macular degeneration, including transpupillary thermotherapy. PMID- 12752046 TI - Telemedicine in ophthalmology. AB - OBJECTIVES: To give an overview of telemedical applications in ophthalmology and to provide background information on new tele-ophthalmological applications. METHODS: We carried out a literature review, a database search and an Internet search. RESULTS: According to published research, the cost-efficiency of telemedicine in ophthalmology has not been established. It has been found to have educational benefits and patients have been satisfied with the possibility of obtaining specialist care without having to travel. CONCLUSIONS: Most studies have been pilot studies and telemedicine is still seldom the primary mode of operation. Technical problems have not been significant, but many open questions about organizational and operational issues remain. Further studies should be directed towards solving these problems and establishing technical standards. PMID- 12752047 TI - Transpupillary thermotherapy of predominantly occult choroidal neovascularization in age-related macular degeneration with 12 months follow-up. AB - PURPOSE: To perform a phase I/II safety and efficacy study in order to assess the outcome following transpupillary thermotherapy (TTT) for occult choroidal neovascularization (CNV) with no or minimally classic CNV in age-related macular degeneration. METHODS: The study comprised 113 referred patients (79 females, 34 males) aged 59-89 years (mean 77.7 years) with predominantly occult CNV. There were 49 cases of occult with no classic CNV and 64 of occult with minimally classic CNV. According to their greatest linear dimension, lesions were classified as being < 3.0 mm or > 3.0 mm. Transpupillary thermotherapy was delivered with a diode laser, emission at 810 nm, duration 60 seconds, laser power 500-800 mW. Clinical examination, ETDRS logMAR visual acuity and fluorescein angiography were carried out at baseline, and at 3, 6 and 12 months in all cases. RESULTS: The average baseline visual acuity was 0.20 (50.6 letters). Following TTT, it was 0.12 (42.0 letters) at 6 months and 0.12 (38.0 letters) at 12 months. Visual acuity improved in 9/113 eyes (8.0%), remained unchanged in 46 (40.7%) eyes, and deteriorated in 58 (51.3%) eyes. There was no significant difference in the proportion of eyes that had lost at least 15 letters at 12 months in the subgroup of occult with no classic CNV (18/49; 36.7%) versus those with minimally classic lesions < 3.0 mm (15/39; 38.5%). However, 13/25 (52.0%) of cases with minimally classic lesions > 3.0 mm had lost at least 15 letters at 12 months (p = 0.31). The most common complications in the 46 eyes that suffered visual loss comprised subretinal progressive fibrosis (18 eyes) and atrophy of the retinal pigment epithelium (13 eyes). CONCLUSIONS: This study shows that TTT generally prevents moderate and severe visual loss at 12 months follow-up in occult CNV with no classic CNV. Eyes with minimally classic lesions with a greatest linear dimension of < 3.0 mm also show the same positive outcome. These results compare favourably with published data on the natural course of the disease. However, minimally classic lesions of > 3.0 mm responded poorly in this treatment setting. PMID- 12752048 TI - Visual perception during phacoemulsification cataract surgery under topical and regional anaesthesia. AB - PURPOSE: To compare the subjective visual experiences of patients during phacoemulsification and intraocular lens (IOL) implantation using regional and topical anaesthesia. DESIGN: A prospective, cohort, questionnaire-based study. METHODS: The study cohort consisted of 247 patients without pre-existing ocular pathology who underwent routine phacoemulsification and IOL implantation. The mean age of the subjects was 75.4 +/- 9.4 years and 34.5% of them had a history of cataract surgery. Three different methods of local anaesthesia were used: 66 (26.7%) of the patients were given topical anaesthesia (TA); 74 (30.0%) were given sub-Tenon's anaesthesia (SA), and 107(43.3%) were given peribulbar anaesthesia (PA). The patients were interviewed immediately after surgery by theatre staff using a standardized questionnaire that investigated their intraoperative visual experiences. RESULTS: There was no significant difference between the three methods of anaesthesia regarding light perception during the surgery. However, patients undergoing surgery under TA experienced brighter light intensity levels (78.3%) than those given SA (50.0%) and PA (55.7%) (p = 0.02). A total of 69.6% of subjects who received TA reported visual perception of colours during surgery, as opposed to 56.8% of SA recipients and 49.0% of PA recipients (p = 0.02). In addition, patients under TA were more aware of surgical instruments (26.1%) than those under SA (10.8%) and PA (15.9%) (p = 0.08). The vast majority of patients in all three groups found the visual experience to be non-frightening. There were no associations between intraoperative visual impression and age or sex. Although not statistically significant (p = 0.06), prior cataract surgery appeared to alleviate some of the anxiety associated with the visual experience. CONCLUSION: Patients undergoing regional and topical anaesthesia experience a wide variety of visual sensations during surgery. The differences in visual impressions between the groups may reflect the varying degrees of optic nerve blockade that result from the different anaesthetics. PMID- 12752049 TI - Refractive errors and visual impairment in 900 adults with intellectual disabilities in the Netherlands. AB - PURPOSE: To collect data on refractive errors and visual impairment in adults with an intellectual disability (ID) in the Netherlands. PATIENTS: A randomized sample of 2100 participants was drawn from a base population of 9000 adults with intellectual disabilities in the Netherlands. This article reports on the first 900 participants. METHODS: All participants underwent a protocol-based on-site ophthalmological assessment carried out by skilled investigators. RESULTS: Co operation was classified according to the number of tests that could be carried out reliably and was good or excellent in 80% of subjects, average in 13% and poor in 7%. Refraction could be reliably assessed in 505/900 (56%) subjects. There was an increased risk of visual impairment in all subgroups compared to the general Dutch population. Visual acuity (VA) was related to the level of ID, but refractive errors were not. New spectacles were prescribed in 106 cases (12%). Of 374 people in whom both monocular VA and the refractive error of the right eye could be reliably assessed, 153 (41%) had a pretest prescription, 16 (10%) of which we found to be inadequate. Of the 221 participants without a pretest prescription, 41 (19%) benefited from correction. Only 38/84 (45%) subjects aged 50 years or older, who could benefit from correction for near vision, had near spectacles. New correction increased the mean distant VA significantly from 0.44 to 0.65 (p < 0.0005). CONCLUSIONS: With some adaptations, visual screening is feasible in a majority of adults with ID. Visual impairment and refractive errors are much more prevalent in adults with ID than in the normal population. Accurate spectacle correction resulted in significant improvement in distant VA. PMID- 12752050 TI - Optic nerve hypoplasia in cholestatic infants: a multiple case study. AB - PURPOSE: To present four infants with optic nerve hypoplasia and cholestasis. METHODS: All patients underwent detailed ophthalmological and hepatological assessment. Their endocrinological and neuroradiological examinations were re evaluated. RESULTS: All four infants presented with cholestasis and were subsequently found to have optic nerve hypoplasia. One child was blind according to the WHO definition and two had low vision. The fourth child had unilateral optic nerve hypoplasia and was too young to be assessed with optotypes. All four children had central nervous system and/or endocrine dysfunction. However, only one child had septo-optic dysplasia. The longterm outcome of liver disease seemed favourable in all children. CONCLUSION: Early assessment by a paediatric ophthalmologist as well as a multidisciplinary approach is of great importance in cholestatic infants. PMID- 12752051 TI - Negative social bias against children with strabismus. AB - PURPOSE: To determine whether the presence of noticeable strabismus creates a negative social bias against children. METHODS: Photographs of two boys and two girls were digitally altered to create photographs of the same child in orthotropic, esotropic and exotropic states. Elementary school teachers rated their perceptions of the children's personal characteristics (using a 10-item list of personal characteristics) based on their responses to these whole-face photographs. The participants were also asked to answer five questions related to the first impressions created by the photographs. RESULTS: Children with esotropia and exotropia were rated more negatively than orthotropic children on all 10 characteristics. Children with esotropia were rated more negatively than those with exotropia on most of these characteristics. CONCLUSIONS: Children with noticeable strabismus are viewed negatively. Hence, correction of strabismus may provide psychosocial benefits even when there is no hope of improving visual function. PMID- 12752052 TI - Ultrasonically measured horizontal eye muscle thickness in thyroid associated orbitopathy: cross-sectional and longitudinal aspects in a Danish series. AB - PURPOSE: To analyse horizontal extraocular muscle findings by ultrasound and exophthalmometry in a tertiary endocrinology centre series of patients with thyroid associated orbitopathy (TAO). METHODS: The 90 thyroid patients included underwent ultrasonic measurement of horizontal eye muscle thickness by a B-scan based technique carried out in addition to their general ophthalmic evaluation. As an indicator of mainly advanced TAO, longterm prednisone or cyclosporine A was given to many of the patients, and drug-resistant visual loss indicated decompression surgery in four of the 90 patients. Thirty-four patients underwent repeated muscle recordings over 15-49 months; this allowed for cross-sectional analysis and the outlining of longitudinal trends. RESULTS AND CONCLUSIONS: (A) Although marginally overlapping, all four muscle groups were significantly thicker in the study group than in normal control subjects. The mean of the sum of all four muscles was 16.8 mm (range 13.6-21.7 mm) in the control group versus 22.6 mm (range 15.5-36.4 mm) in the thyroid group. (B) Using the clinical NOSPECS grading, more advanced eye involvement was found to generally result in a higher exophthalmometric measurement of protrusion and eye muscle thickness. However, slender rectus muscles and/or normal exophthalmometric values might occur even in advanced orbitopathy. (C) Over a period of 2-4 years, only a few of 34 patients with satisfactory serial ultrasonic measurements returned to their premorbid ophthalmic status. Typically, the extraocular muscles kept their abnormal size after having become clinically quiescent (fibrotic). (D) We found no safe indication regarding disease stage, active or late, from the ultrasonic appearance of the muscle tissue. (E) Discrepancies between various normative eye muscle studies are discussed with regard to computer tomography and magnetic resonance imaging. PMID- 12752053 TI - A new technique for removing the inner wall of Schlemm's canal using cyanoacrylate. AB - PURPOSE: To introduce a new technique for removing the inner wall of Schlemm's canal using cyanoacrylate. METHODS: Two donor eyes were used in this study. A limbal-based scleral flap was dissected. The external wall of Schlemm's canal was opened and the inner wall of Schlemm's canal was exposed. Ethyl-2-cyanoacrylate was applied uniformly to the inner wall. After it had solidified, it was removed with fine forceps. The operated sites were then studied by light microscopy. RESULTS: After cyanoacrylate removal, the aqueous humour was seen percolating through the thin remaining trabecular meshwork. Histopathological findings at the operated sites revealed that the lining of the Schlemm's endothelial cells was irregular. Endothelial cell nuclei were rarely seen along the canal wall. Microperforation to the anterior chamber was not seen. CONCLUSION: This new technique is potentially a safer method for removing the inner wall of Schlemm's canal than the conventional method. PMID- 12752054 TI - The effects of latanoprost and brimonidine on blood flow velocity of the retrobulbar vessels: a 3-month clinical trial. AB - OBJECTIVE: To investigate the effects of topical latanoprost 0.005% and topical brimonidine tartrate 0.2% on retrobulbar blood flow in patients with primary open angle glaucoma (POAG) and ocular hypertension (OHT). METHODS: Forty-one consecutive patients with POAG and OHT were enrolled in this prospective, open label, randomized, parallel study. Patients received either latanoprost 0.005% or brimonidine 0.2% for 3 months. Baseline retrobulbar blood flow measurements of the ophthalmic artery, central retinal artery and temporal short posterior ciliary artery were taken using colour Doppler imaging ultrasound, concurrently with systemic blood pressure, heart rate, ocular perfusion pressure and intraocular pressure (IOP) measurements. These measurements were repeated after 3 months. RESULTS: Both latanoprost and brimonidine significantly reduced IOP (p < 0.05). While there was a statistically significant increase in peak systolic velocity of the ophthalmic artery, no significant change was observed in the other vessels with latanoprost treatment (p < 0.05). Topical brimonidine did not significantly alter flow velocities or resistive indices in the retrobulbar vessels after 3 months. CONCLUSION: Topical latanoprost and brimonidine significantly reduced IOP in patients with POAG and OHT without causing significant haemodynamic alterations in the retrobulbar vessels. PMID- 12752055 TI - The relationship between serum trace element changes and visual function in heavy smokers. AB - PURPOSE: The aim of this prospective study was to evaluate serum manganese (Mn), zinc (Zn) and copper (Cu) levels and visual functions including visual acuity, colour vision, pattern visual evoked potentials (PVEPs), and contrast sensitivity in heavy smokers and to compare these with the equivalent levels and functions in non-smokers. METHODS: Data were recorded in 24 healthy, chronic and heavy cigarette smokers and 16 healthy, non-smoking control subjects. Serum Zn, Cu and Mn concentrations in all subjects were measured by atomic absorption spectrophotometry. RESULTS: Both study and control subjects had normal visual acuity and colour vision. Pattern visual evoked potentials were normal in all study and control subjects. Contrast sensitivity was significantly reduced in heavy smokers compared to non-smokers (p < 0.023), despite the fact that central vision and PVEP responses were not affected. Mean serum Mn and Zn levels were significantly lower in smokers than in non-smokers (p < 0.0001 and p < 0.005, respectively). CONCLUSIONS: Reduced contrast sensitivity values associated with low levels of serum Mn and Zn, which function as cofactors of superoxide dismutase in erythrocyte and other nucleated cells, suggest a possible role of trace elements in smoking-induced early retinal toxicity. PMID- 12752056 TI - Maximum acceptable dose of ultraviolet radiation: a safety limit for cataract. AB - PURPOSE: To develop a method for experimental estimation of toxicity for continuous dose-response relationships. To apply this method to cataract induced by ultraviolet radiation (UVR) in young rats. METHODS: After establishing experimentally the frequency distribution of light scattering of normal physiologically clear lenses, the lower limit of pathological light scattering is defined such that a certain fraction, for example 97.5%, of normal lenses scatter less light. RESULTS: The dose-response function for UVR and cataract is determined experimentally. With this function, the dose corresponding to the lower limit of pathological light scattering may be determined as the maximum acceptable dose (MAD). The MAD0.975 for UVR 300 nm was determined to be 2.2 kJ/m2. CONCLUSIONS: The method can serve as a basis for establishing safety standards for UVR-induced cataract and probably other continuous dose-response functions. PMID- 12752057 TI - Growth of cultured porcine retinal pigment epithelial cells. AB - PURPOSE: To establish and characterize cultures of porcine retinal pigment epithelial (pRPE) cells in order to produce confluent monolayers of cells for transplantation. METHODS: Primary pRPE cell cultures were established. Cell morphology was assessed by phase contrast and electron microscopy. Growth was determined by the crystal violet dye uptake assay. DNA synthesis and content were measured by incorporation of 3H-thymidine and flow cytometry. RESULTS: This primary culture resulted in cells with well-preserved morphology that could be propagated in up to six passages. The deterioration observed over time in cultures was not due to a constant high rate of cell turnover as postconfluency cell proliferation was limited. However, a large fraction of the cells had a high DNA content despite a lack of active DNA synthesis. CONCLUSIONS: The present method yields pRPE cells of high purity and proliferative capacity with preserved epithelial phenotype. However, aberrant DNA profiles and the deterioration of cell morphology observed over time in this graft material represent serious problems in RPE transplantation. PMID- 12752058 TI - Effects of topical vitamin E on corneal superoxide dismutase, glutathione peroxidase activities and polymorphonuclear leucocyte infiltration after photorefractive keratectomy. AB - PURPOSE: Photorefractive keratectomy (PRK) induces free radical formation and polymorphonuclear (PMN) cell infiltration in the cornea. Vitamin E is a free radical scavenger and protects the cells from reactive oxygen species. We investigated the effects of topical vitamin E on corneal PMN cell infiltration and corneal antioxidant enzyme activities after PRK. METHODS: We studied four groups, each consisting of seven eyes. Group 1 were control eyes. In group 2 the corneal epithelium was removed by a blunt spatula (epithelial scrape). In group 3, corneal photoablation (59 micro m, 5 dioptres) was performed after epithelial removal (traditional PRK). In group 4 we tested the effects of topical Vitamin E after traditional PRK. Corneal tissues were removed and studied with enzymatic analysis (measurement of corneal superoxide dismutase and glutathione peroxidase activities) and histologically. RESULTS: Stromal PMN leucocyte counts were significantly higher after mechanical epithelial removal and traditional PRK (p < 0.05). Corneal superoxide dismutase and glutathione peroxidase activities decreased significantly after mechanical epithelial removal and traditional PRK (p < 0.05). In group 4, treated with vitamin E, corneal superoxide dismutase activity did not differ significantly from that in the medically non-treated groups, nor did corneal PMN cell infiltration after traditional PRK. The reduction of corneal glutathione peroxidase activity after PRK was reduced significantly after topical vitamin E treatment. CONCLUSIONS: Topical vitamin E treatment may be useful for reducing the harmful effects of reactive oxygen radical after epithelial scraping and PRK in that it increases corneal glutathione peroxidase activity. PMID- 12752059 TI - The effect of vitamin E on stress-induced changes in visual evoked potentials (VEPs) in rats exposed to different experimental stress models. AB - PURPOSE: The aim of the study was to investigate the effects of vitamin E on stress-induced changes in visual evoked potentials (VEPs) and lipid peroxidation. METHODS: Eight experimental groups of 10 rats per group were formed. These consisted of the control group (C); the group treated with vitamin E (E); groups exposed to cold stress (CS), immobilization stress (IS) and both cold and immobilization stress (CIS), and groups exposed to equivalent stresses and treated with vitamin E (CSE, ISE, CISE). Vitamin E was injected intramuscularly in a dose of 30 mg/kg/day. RESULTS: Following chronic stress (15 days), plasma corticosterone concentrations in all experimental groups were significantly increased over those in C group. Vitamin E significantly decreased corticosterone levels in all stress groups compared with their respective control groups. Brain nitrite levels were significantly more elevated in all stress groups than in the C group. Vitamin E reduced retina and brain nitrite levels in all stress and E groups compared with their respective control groups. Vitamin E decreased glutathione peroxidase (GSH-Px) activity in retina and brain tissues in the CSE group, but increased it in the ISE group compared with their respective control groups. Lipid peroxidation was increased in brain and retina tissues in all stress groups as indicated by the significant increase in thiobarbituric acid reactive substance (TBARS) levels with respect to the C group. Vitamin E produced a significant decrease in brain and retina TBARS levels in all stress groups with respect to their corresponding control groups. The mean latencies of P1, N1, P2, N2 and P3 components were significantly prolonged in all stress groups compared with the C group. CONCLUSION: Vitamin E returned the VEP latencies in the stress groups to control values. Our findings clearly indicated that vitamin E has the potential to prevent VEP changes caused by stress. PMID- 12752060 TI - Filtration bleb after phacoemulsification. PMID- 12752061 TI - Surgical treatment of symptomatic primary pupillary pigment epithelial iris cysts. PMID- 12752062 TI - Topical alpha-interferon in recurrent conjunctival papilloma. AB - PURPOSE: To report the effect of topical alpha-interferon (alpha-IF) in two cases of recurrent conjunctival papillomas. METHODS: One patient with a conjunctival papilloma and one patient with a limbus-based papilloma were treated with excision and cryotherapy. Recurrences included one in situ carcinoma. Retreatment included systemic and topical alpha-IF. RESULTS: In Case 1, the papillomas regressed with systemic alpha-IF followed by topical application of alpha-IF (2.8 x 106 U) for 1 year against recurrence. In Case 2, the papillomas disappeared with topical alpha-IF over 1 year. No recurrences were seen during follow-up (84 and 91 months, respectively). The possible side effect of topical alpha-IF was superficial keratitis. Retrospective polymerase chain reaction tests for HPV were positive and showed type 6 in Case 1 and type 33 in Case 2. CONCLUSION: Topical alpha-IF can be used as adjuvant therapy in recurrent conjunctival papillomas. PMID- 12752063 TI - Overtreatment of transpupillary thermotherapy for choroidal neovascularization. PMID- 12752064 TI - Hemi-retinal artery occlusion associated with sexual activity and sildenafil citrate (Viagra). PMID- 12752065 TI - Bilateral retinal macroaneurysms with neurofibromatosis type 1. PMID- 12752066 TI - Bilateral centripetal lipid keratopathy with diffuse anterior scleritis. PMID- 12752069 TI - When and how should bone mineral density be measured? PMID- 12752070 TI - Elective cervical cerclage for prevention of preterm birth: a systematic review. AB - BACKGROUND: Elective cervical cerclage has been purported to prevent spontaneous preterm birth. We present a systematic review to determine the effectiveness of cervical cerclage in preventing spontaneous preterm birth before 34 weeks' gestation. METHODS: Searches were conducted in MEDLINE, EMBASE, Cochrane Library, and Science Citation Index to identify randomized trials published between 1966 and 2002. All randomized trials that evaluated the effectiveness of elective cerclage compared with no cerclage in women who were at risk of preterm birth before 34 weeks' gestation were included for analysis. Quality assessment and data extraction were performed in duplicate. RESULTS: There were seven relevant trials, comprising 2354 women. Meta-analysis was inappropriate because of large differences in the quality of the studies. However, in the largest single trial of good quality, cerclage was shown to prevent birth before 34 weeks' gestation. In this single study the reported number to be treated to prevent one additional preterm birth before 34 weeks was 24 women (95% CI: 10-61). The results of other trials were consistent with the finding of the largest trial. Data on complications were sparse and inconclusive. CONCLUSION: Our systematic review shows that elective cervical cerclage has a significant effect in preventing spontaneous preterm birth before 34 weeks' gestation. Further research should focus on identification and quantification of possible complications, and of risk factors and tests that identify high-risk women who would benefit most from cerclage. PMID- 12752071 TI - Menorrhagia: an update. AB - Menorrhagia is defined as a 'complaint of heavy cyclical menstrual bleeding occurring over several consecutive cycles'. Objectively it is a total menstrual blood loss equal to or greater than 80 ml per menstruation. It is estimated that approximately 30% of women complain of menorrhagia. Excessive bleeding is the main presenting complaint in women referred to gynecologists and it accounts for two-thirds of all hysterectomies, and most of endoscopic endometrial destructive surgery. Thus, menorrhagia is an important healthcare problem. Its etiology, investigation, medical and surgical management are described. In approximately 50% of cases of menorrhagia no pathology is found at hysterectomy. Abnormal levels of prostaglandins or the fibrinolytic system in the endometrium have been implicated. Effective medical treatments suitable for long-term use include intrauterine progestogens, antifibrinolytic agents (tranexamic acid) and nonsteroidal anti-inflammatory agents (mefenamic acid). Over the past decade there has been increasing use of endometrial destructive techniques as an alternative to hysterectomy. Their further refinement and the advent of fibroid embolization has increased the options available to women. PMID- 12752072 TI - Microbial invasion and cytokine response in amniotic fluid in a Swedish population of women with preterm prelabor rupture of membranes. AB - BACKGROUND: Previous studies have shown an association between intra-amniotic microbial invasion and/or inflammation and spontaneous preterm birth. The aim of this study was to investigate the occurrence of intra-amniotic microorganisms and cytokines [interleukin (IL)-6 and IL-8] in a Swedish population, with low incidence of preterm birth, of women with preterm prelabor rupture of membranes and their correlation to preterm birth. METHODS: Amniotic fluid was retrieved transabdominally from 58 patients with preterm prelabor rupture of membranes before 34 weeks of gestation. Polymerase chain reaction (PCR) analyses for Ureaplasma urealyticum and Mycoplasma hominis and culture for aerobic and anaerobic bacteria were performed. IL-6 and IL-8 were analyzed with enzyme-linked immunosorbent assay (ELISA). RESULTS: Microorganisms in amniotic fluid were detected in 13 patients (25%). Patients with bacteria detected in the amniotic fluid had significantly higher levels of IL-6 and IL-8. An amniotic fluid concentration of IL-6 >/= 0.80 ng/ml [relative risk 1.93, 95% confidence interval (CI) 1.13-3.29, sensitivity 63%, specificity 75%] was associated with an increased risk of delivery within 7 days. There was also an association between IL-8 and preterm birth (< 34 weeks). CONCLUSIONS: Intra-amniotic microbial invasion and inflammation in this population of Swedish women with preterm prelabor rupture of membranes were similar to data reported from populations with a higher incidence of preterm delivery. Amniotic IL-6 correlated to the presence of microorganisms and delivery within 7 days and IL-8 to delivery before 34 weeks. PMID- 12752073 TI - Influence of leptin and the TNF system on insulin resistance in pregnancy and their effect on anthropometric parameters of newborns. AB - BACKGROUND: We studied the contribution of the tumor necrosis factor system and leptin to insulin resistance during the course of normal pregnancy. METHODS: Forty-five healthy pregnant women (15 in the 1st, 15 in the 2nd2 and 15 in the 3rd3 trimester) and 25 age-matched healthy nonpregnant women as controls. Twenty three newborns delivered by women followed in the 2nd and 3rd trimesters were also investigated. Fasting serum immunoreactive tumor necrosis factor-alpha, soluble tumor necrosis factor receptor-1, soluble tumor necrosis factor receptor 2, leptin (by enzyme-linked immunoassay) and C-peptide (by radioimmunoassay) concentrations in the patients and controls were measured. Body weight, length and head circumference of the newborns were analyzed in connection with the measured maternal parameters. RESULTS: Significantly elevated tumor necrosis factor-alpha, tumor necrosis factor receptor-1 and -2, leptin, and C-peptide levels were found in the 3rd3 trimester as compared with the 1st1 and 2nd2 trimesters and with the nonpregnant controls. Tumor necrosis factor-alpha, tumor necrosis factor receptor-2, C-peptide, leptin concentrations and body mass index were found to be in a significant positive linear correlation with each other. Significant negative linear correlations were calculated among maternal serum C peptide, tumor necrosis factor-alpha and leptin concentrations and the head circumference of the newborns. CONCLUSIONS: In conclusion, increased tumor necrosis factor-alpha and leptin levels may contribute to insulin resistance in late pregnancy. Tumor necrosis factor-alpha and leptin may be regulators of intrauterine bone development of newborns. PMID- 12752074 TI - Comparison between two cord blood collection strategies. AB - BACKGROUND AND OBJECTIVES: Collection strategy is the first step for collecting good quality cord blood (CB) units. There are two principal different techniques to collect CB from the umbilical vein: in the delivery room while the placenta is still in the uterus by midwives and obstetricians or in an adjacent room after placental delivery by CB-bank trained personnel. In this study, the benefits and disadvantages between two different CB collection strategies were evaluated in order to improve CB bank methodology. DESIGN AND METHODS: Valencia CB bank maintains the two different collection strategies aforementioned. Before processing CB units, volume was calculated and samples were drawn for cell counts. After processing and before cryopreservation, samples for cell counts, CD34 analysis, viability, clonogenic assays and microbiology were drawn directly from the bags. We compared the efficiency of the two collection techniques. RESULTS: Obstetric date and umbilical CB was obtained from 848 vaginal (484 collected in uterus and 364 collected ex uterus). The proportion of excluded CB units before processing was 33% for ex uterus and 25% for in uterus. The difference was statistically significant. A larger volume and a higher number of total nucleated cells, CD34+ cells and CFUs were harvested in the in uterus collection group. INTERPRETATION AND CONCLUSIONS: Based on our findings, we conclude that the mode of collection influences the hematopoietic content of CB donations. Collection before placental delivery is the best approach to CB collection and allows optimizing CB bank methodology. PMID- 12752075 TI - Feasibility of simultaneous application of fetal electrocardiography and fetal pulse oximetry. AB - BACKGROUND: Fetal pulse oximetry measures arterial oxygen saturation during delivery. The fetal electrocardiogram STAN S21 analyzes the repolarisation segment of ECG (ST) waveform, which is altered by the intramyocardial potassium release resulting from metabolic acidemia. This study aimed to evaluate the feasibility of a simultaneous application of pulse oximetry and fetal electrocardiography and to estimate any agreement between both methods indicating fetal compromise. METHODS: In an observational trial 35 fetuses were simultaneously monitored by pulse oximetry (OBS-500) and electrocardiography. The evaluation focused on signal output and on the coincidence of desaturation in fetal pulse oximetry and on ST events. Desaturation was defined as a drop of at least 20% of oxygen saturation from base line level occurring within 1 min (steep desaturation) or duration of time (s) with oxygen saturation below the threshold of 30%. Statistical analysis was performed with the Mann-Whitney U-test for continuous variables. RESULTS: Signal output of the simultaneous application of both sensors was not significantly reduced in the electrocardiogram and in pulse oximetry (8% vs. 12% reduction). In 15 of the 35 fetuses, ST events indicating fetal hypoxia occurred. In these cases, pulse oximetry showed significantly more episodes of desaturation in comparison with fetuses without ST events. Median saturation during the ST events was significantly lower than in the recordings without ST events (60% vs. 74%, p < 0.05). In the umbilical artery these neonates showed significantly lower pH (7.19 vs. 7.33, p < 0.001), significantly higher lactate (5.1 vs. 3.4 mmol/l, p < 0.05) and significantly lower base deficit ( 9.4mmol/l vs. -4.0 mmol/l, p < 0.001) levels. CONCLUSIONS: The combination of fetal pulse oximetry and fetal electrocardiography appears feasible and indicates signs of intermittent hypoxia. These findings may encourage the development of technology that combines these different methods of monitoring. PMID- 12752076 TI - Involvement of the glycoproteic meshwork of cervical mucus in the mechanism of sperm orientation. AB - BACKGROUND: The aim of this study was to investigate the influence of the solid phase of the mucus hydrogel in facilitating the upward movement of spermatozoa during the menstrual cycle. METHODS: A total of 171 sperm migration experiments using 47 selected ovulatory mucus samples were performed in vitro in tubes of various shape and diameter. Physical constraints applied to cervical mucus combined with scanning electron microscopy (SEM) techniques permitted analysis of the mechanism of sperm orientation within cervical mucus submitted to capillary constraints simulating the various conditions of sperm orientation in vivo. RESULTS: The results support the theory according to which spermatozoa entering the cervical canal are constrained to follow the oriented micellar lines of strain. Stretched ovulatory mucus exhibits noticeable prevalent orientation of most filaments aligned in a manner roughly parallel to the traction axis. By contrast, even after strong stretching applied to luteal-type mucus and longer contact periods, the alignment of glycoproteic meshes was not sufficiently marked to allow spermatozoa to orient themselves and to move rapidly. The results are discussed as a function of the variations occurring in the three-dimensional macromolecular arrangement of the glycoproteic framework subjected to flow constraints. CONCLUSIONS: The study confirms that there is a close relationship between the three-dimensional arrangement of the mucus framework and the ability of spermatozoa to move rapidly in a given direction. The high resolution and three-dimensional aspect of SEM micrographs support the correlation of sperm orientation with current data on rheologic properties of biologic hydrogels. PMID- 12752077 TI - Embryo transfer by midwife or gynecologist: a prospective randomized study. AB - BACKGROUND: Embryo transfer (ET) in assisted reproduction treatments has traditionally been performed by gynecologists in the Nordic countries. As gynecologists often have a busy schedule, midwives and nurses have become increasingly important in performing the treatment, providing subject information, ultrasound monitoring and assistance at ET. As part of the continuous development of our IVF treatment we have carried out a prospective randomized pilot study where either a midwife or a gynecologist has performed ET. The aim of this study was to see if a skilled IVF midwife could perform ET with similar results to a gynecologist. METHODS: On the day of oocyte aspiration the subjects were randomized, by means of closed envelopes, for ET to be performed either by a midwife or a gynecologist. A total of 102 subjects were included in the study, 51 for ET by a skilled midwife and 51 by a gynecologist. There were no differences in the groups in respect to ET routine and catheters used. RESULTS: No significant differences were observed in subject characteristics as regards age, method of pituitary down-regulation or proportion of IVF/ICSI cycles. Similar clinical pregnancy rates between ETs performed by midwives vs. gynecologists, 31% vs. 29%, respectively, were seen. Subject experience as judged by a questionnaire also showed high acceptance of ET by a midwife. CONCLUSION: The results show that it is a feasible option to allow midwives to carry out ETs. PMID- 12752078 TI - Reactions to awareness of activated protein C resistance carriership: a descriptive study of 270 women. AB - BACKGROUND: Around 25 million Caucasian women are carriers of the FV Leiden mutation that causes activated protein C (APC) resistance. This is a heritable condition with a lifelong increased risk of venous thromboembolism. We performed this study to investigate women's reactions to their awareness of being APC resistant and the consequences of this awareness. METHODS: All APC-resistant women (n = 270) included in a prior study on APC resistance and pregnancy (n = 2480) were invited by written questionnaire to describe their reactions to having APC resistance, how this had changed their lives, and how they experienced our information. Answers were obtained from 215 of the 270 women (80%). RESULTS: More than 94% of the APC-resistant women were satisfied with knowing themselves to be APC-resistant and pleased that they had enrolled in the study. Of the women on combined oral contraceptives (COC), 84% changed their method of contraception, but 16% continued on COC. One-third of the women reported becoming more worried or afraid of getting pregnant again as a result of their awareness of being APC resistant. The proportion of women who sought legal abortions during a 2-year period after receiving this information was similar in both subgroups: 4.4% (12/270) vs. 4.3% (94/2210), p = 0.9. CONCLUSIONS: We conclude that most APC resistant women were pleased to learn of their APC resistance status, that there was not an increased incidence of legal abortions, but almost one-third reported being more worried or afraid of getting pregnant again. PMID- 12752079 TI - Anatomic and functional assessment and risk factors of recurrent prolapse after vaginal sacrospinous fixation. AB - BACKGROUND: To identify risk factors in recurrence and to evaluate anatomic and functional results of vaginal sacrospinous ligament fixation and pelvic floor reconstruction for genital prolapse. METHODS: One hundred and thirty-eight women underwent surgery for uterovaginal or vault prolapse. Follow-up data were available for 122 cases; 83% were examined and others were interviewed by telephone. The median (range) follow-up was 24 (1-141) months. Cox regression was used to identify risk factors associated with recurrence; uni- and multivariate regression was used to identify risk factors underlying postoperative infections because infections were found to be a risk factor of recurrence. Recurrence-free survival was estimated using the Kaplan-Meier method. RESULTS: Seven (5%) patients suffered severe cardiopulmonary complications including one postoperative death due to a pulmonary embolism. Twenty-six (21%) patients suffered a recurrence, 14 with cystocele. Ten patients with recurrence were symptomatic and six underwent a re-operation. The Cox regression model showed that vaginal cuff infection raised the odds ratio (OR) for recurrence to 6.13 [confidence interval (CI) 1.80-20.83] and urinary tract infection to 3.65 (CI 1.40-9.47). In both uni- and multivariate analysis, lack of intravenous antibiotic prophylaxis, age less than 73 years and vaginal ulcerations were statistically significant risk factors for postoperative infection. Eleven (33%) out of 33 sexually active women reported improvement and three (9%) complained of dyspareunia. CONCLUSIONS: Transvaginal sacrospinous ligament fixation with pelvic floor repair is an effective means of correcting both vault prolapse and uterine procidentia. Women who wish to preserve coital function will also benefit from this operation. Postoperative infection is an independent and most important individual risk factor underlying recurrence. Prophylactic antibiotics seem to be effective in reducing the rate of postoperative infections. PMID- 12752080 TI - Acute massive fetomaternal hemorrhage: case reports and review of the literature. PMID- 12752081 TI - Urachal myoma: a case report. PMID- 12752082 TI - Aggressive angiomyxoma of the female perineum in pregnancy. PMID- 12752083 TI - Primary retro peritoneal mucinous cystadenoma. PMID- 12752084 TI - Catamenial pneumothorax treated by laparoscopic tubal occlusion using Filshie clips. PMID- 12752085 TI - [New design concept to improve medical application software]. AB - Up to now, there has been no software model available that adequately addresses the growing importance of flexibility in using different information tools. Based on Java, Jini technology provides distributed and therefore robust software architecture. In case of an application crash, Jini is able to minimize the damage by adding functions that reverse the effects of the crash. Owing to increasing user mobility, it is necessary to be able to receive location independent information. The growing use of powerful telecommunications suggests the application of 'wireless application protocol' (WAP) mobile telephones also for medical purposes. This paper presents the application of these new software trends (Jini and wireless application protocol). PMID- 12752086 TI - Monitoring the efficacy of iodine-131-MIBG therapy using fluorine-18-FDG-PET. AB - BACKGROUND: The purpose of this study was to assess the potential of fluorine-18 fluoro-2-deoxy-d-glucose positron emission tomography (18F-FDG-PET) for monitoring the efficacy of iodine-131 metaiodobenzylguanidine (131I-MIBG) therapy in neuroendocrine tumours. METHODS: A total of seven 131I-MIBG therapies with 3.7 to 10.2 GBq were carried out in three patients suffering respectively from a phaeochromocytoma, a paraganglioma and a metastatic neuroendocrine tumour of an unknown primary. The post-therapeutic whole-body scintigrams were compared with the results of six 18F-FDG-PET studies performed at the time of the therapies. One patient received three PET scans prior to each one of the MIBG therapies, and one patient was studied twice. RESULTS: 18F-FDG uptake in tumour sites seemed to correlate well with tumour differentiation, showing no uptake in one patient with a highly differentiated neuroendocrine tumour, and moderate-to-intense uptake in the two other patients with metastatic disease. Those tumour sites that had a simultaneous positive uptake in both the MIBG scintigram and the PET scan showed response to therapy as a continuous reduction in MIBG uptake over time. They also showed a qualitative decrease in FDG accumulation during the follow-up. This was associated with a decrease in the mean and maximum standard uptake values of more than 50 % in some metastases, while the X-ray computed tomography showed no decrease in tumour volume. Two patients revealed additional metastases that were unknown on the basis of prior diagnostic or therapeutic PET scans and radiological follow-up. CONCLUSIONS: It may be concluded from these cases that 18F-FDG-PET is a valuable tool for an initial metabolic staging of neuroendocrine tumours prior to 131I-MIBG therapy, as it can reveal tumour sites beyond the reach of radioisotope therapy. It may also be of importance in assessing therapeutic potential in those tumour sites that do show positive MIBG uptake. PMID- 12752088 TI - Trends in primary health care consumption by the elderly. AB - Trends in primary health care consumption were monitored in the elderly (60 or more years of age). Respondents (5162) were picked by random mechanical selection. They were monitored for a complex of characteristics composed of frequency of visits to a GP office, frequency of procedures performed at the GP office, drug prescriptions, frequency (and type) of procedures performed at specialists' offices, and hospitalization. The study was conducted in 1982, 1992, 1993 and 1996. Results were statistically analysed. There was a statistically significant increase in the consumption of primary health care by the elderly: GP office visits rose from 80 % to 98.8 %, and there was a quantitative change in procedure types. There was also a marked increase in drug prescription: from 77 % receiving prescriptions in 1982 to 97 % in 1996. The highest consumption was of cardiotonics, analgesics and antipyretics, which corresponds to an increasing prevalence of chronic diseases and multimorbidity. The results of our studies underline the important role of primary care physicians in the care of the elderly, who represent 80 % of the clientele of their offices in our country. PMID- 12752087 TI - Accuracy of whole-body 18F-FDP-PET for restaging malignant lymphoma. AB - BACKGROUND: The aim of this retrospective study was to evaluate the accuracy of fluorine-18 fluorodeoxyglucose positron emission tomography (18F-FDG-PET) images, which were interpreted under daily routine conditions, in patients with Hodgkin's disease (HD) or non-Hodgkin lymphoma (NHL) for restaging after chemotherapy and/or radiotherapy. For this purpose, 18F-FDG-PET results were compared with morphological imaging methods and the patients' clinical background. METHODS: 121 PET images of 93 lymphoma patients (44 HD, 49 NHL) were investigated after chemotherapy/radiotherapy. For PET imaging, 160-200 MBq 18F-FDG was administered intravenously, followed by an infusion of 20 mg Furosemid in 250 mL saline. Whole body 18F-FDG-PET images were obtained using a partial-ring PET scanner without attenuation correction. The morphological imaging consisted in computed tomography and ultrasound (CT/US) in all patients, additional MRI in some patients, and iliac crest biopsy in cases of suspicious bone marrow involvement. The standard of reference was composed of biopsy data, clinical status at the time of investigation, and follow-up of at least 12 months. The PET images were evaluated for their sensitivity, specificity and accuracy based on written reports, which were compiled from other imaging data and the clinical history of the patients. RESULTS: Sensitivity, specificity, and accuracy of 18F-FDG-PET was 91 %, 81 %, and 85 %; of CT/US, 88 %, 35 %, 56 %, respectively. Major sources of error in 18F-FDG-PET were due to asymmetric muscular hypermetabolism and inflammatory lesions misinterpreted as persistent viable lymphoma tissue. Furthermore, secondary malignancies other than lymphomas were another reason for misinterpretations of 18F-FDG-PET studies. CONCLUSIONS: 18F-FDG-PET showed a comparable sensitivity but a higher specificity and accuracy compared with CT/US. To achieve a high accuracy in 18F-FDG-PET, the nuclear medicine specialist needs imaging and clinical data as background information, which can only be acquired through close co-operation with the referring clinicians. Pharmacological muscular relaxation in the course of 18F-FDG-PET imaging may be advisable, as nonspecific muscular hypermetabolism was one of the problems at the image readings and a source of incorrect 18F-FDG-PET interpretations. PMID- 12752089 TI - The influence of obesity on the frequency and distribution of medication. AB - Obesity is a serious health problem in industrialized countries and is associated with a significant increase in total health care costs. Only few data are available about the costs of drug therapies in patients with an increased body weight treated under clinical routine procedures. Such data could support efforts to intensify obesity prevention and treatment programmes in order to reduce comorbidities and costs. We have evaluated body mass index (BMI), diagnosis, and medication in 3360 outpatients (2175 women and 1185 men; mean age: 56.7 +/- 17.5 years). All patients underwent physical examinations, including BMI determination, and provided a detailed record concerning medication. In 1809 patients, the percentage of body fat content was measured with a bioimpedance method (OMRON BF 302 body fat monitor). Continuous variables were compared using the t-test or Wilcoxon U-test. Frequency distributions were compared using chi squared tests. With respect to BMI, most of the patients (n = 1793; 53 %) were overweight or obese, 1349 (40 %) showed a normal BMI and 218 (7 %) a low BMI. The majority of cardiovascular (61 %), rheumatological (61.1 %) and metabolic (60.4 %) medication was administered to overweight and obese patients. Parallel findings could be obtained by analysing the percentage of body fat and the frequency of medication. Overall, 82.5 % of all medication was given to patients with a body fat content >20 %. Our results support the importance of weight reduction programmes in order to prevent an overall increase in the costs of medication as a consequence of overweight and obesity. PMID- 12752090 TI - [Muscle cell proteins are selectively released into the blood stream by marathon running]. AB - In 19 marathon runners of both sexes, plasma concentrations of total creatine kinase (CK) activity, CKMB mass, myoglobin and troponin I were determined before and immediately after the race. Total CK activity and myoglobin increased significantly in all runners and showed neither a correlation with the individual age of the runners nor with the time they needed to reach the goal. In 12 of the runners, CKMB mass increased during the race to a level suggesting myocardial necrosis. However, the runners did not show any detectable deterioration of cardiac function after the race. The appearance of considerable amounts of muscle proteins in plasma precipitated by the muscle strain during the race seems explained by damage of skeletal muscle detected by histological studies. These phenomena may also be a consequence of profoundly disturbed cellular permeability, perhaps due to a kind of local stunning of muscle tissue by prolonged muscular strain. PMID- 12752091 TI - Advice against smoking is not effective in patients with Graves' ophthalmopathy. AB - Advice against smoking is one of the means frequently used in the management of patients with Graves' ophthalmopathy (GO). The objective of this study was to prospectively evaluate the efficacy of such advice. The study was performed at a referral centre. Eighty-five smokers out of 102 consecutive patients with GO, who were examined during a 1-year period, were included in the study. Severity of the disease, smoking history, and the results from a patients' self-assessment questionnaire were recorded. The behavioural intervention consisted in a standardized 2-min message from the attending ophthalmologist strongly advising each patient to quit. The main outcome measure was smoking cessation during the 1 year follow-up period. None of the 85 smokers reported smoking cessation either during the follow-up visits or at the end point of the study. The results of our study show that advice against smoking is not effective among patients with GO. Specifically designed intervention programmes may be necessary and should be evaluated in a controlled trial. PMID- 12752092 TI - Unusual localizations of hydatid disease. AB - Hydatid disease is endemic in several Mediterranean countries, posing an important health problem for these countries. The hydatid cyst is characterized by cystic lesions with clear boundaries, which can be observed in all parts of the body. Approximately 70 % of hydatid cysts are situated in the liver, followed by the lung (25 %). The kidneys, spleen, bile ducts, mesentery, soft tissue and brain are less frequent sites. We investigated patients who were treated for hydatid disease in our departments in the last 5 years with respect to localization of the disease, symptoms, surgical intervention, length of hospitalization, diameters of the cyst, and classification by Gharbi. In this retrospective and descriptive study, 176 patients are evaluated who were treated for hydatid disease between 1995 and 1999 in our departments. Of these patients, 14 were included with localization other than in the liver and lungs. Fourteen of the patients diagnosed with unusually located hydatid disease were men, six were women. Their mean age was 41.6 +/- 20.8 years; the length of hospital stay was 7.07 +/- 0.4 days. Overall, 28.6 % of patients with unusually located hydatid cyst had recurrent disease. The time period since last cyst operation was 5.25 +/ 3.5 years. The mean cyst diameter was 96.5 +/- 54.5 mm. According to Gharbi's classification, three cases (21.4 %) of the unusually located hydatid cysts were type I, two (14.3 %) type II, and eight (57.1 %) type III. There was only one case of type IV and no cases of type V. Spleen and kidneys are the organs where hydatid disease is most frequently observed after the liver and lung. It can be observed in all parts of the body including the brain, peritoneum, mesenterium, choledochus, pancreas, bone and muscles. The type of treatment is determined by the localization and type of hydatid disease. Surgical treatment for splenic hydatid cysts is splenectomy. The functional kidney should be saved in non communicable hydatid disease. Total excision is almost never possible; endocystectomy and drainage procedure should be preferred for hydatid disease of the brain, pancreas and choledochus. Chemotherapy is usually given because of the risk of recurrence; this medical treatment consists in albendazole and mebendazole administration for 3-6 months in the postoperative period. PMID- 12752093 TI - Fluorescence in situ hybridization for the BCR-ABL fusion gene in a patient with imatinib mesylate-resistant chronic myelogenous leukaemia in extramedullary blast crisis. PMID- 12752094 TI - Granulocytic sarcoma of main biliary ducts. PMID- 12752095 TI - Heparin-induced thrombocytopenia: pathogenesis and management. PMID- 12752096 TI - Karl Landsteiner and his major contributions to haematology. PMID- 12752098 TI - A prognostic model for survival in chronic lymphocytic leukaemia based on p53 expression. AB - As the abnormal expression of p53 protein is prognostically significant in some human cancers, its significance in patients with B-cell chronic lymphocytic leukaemia (CLL) was assessed. Two investigators evaluated the percentage of bone marrow mononuclear cells that stained for p53, using biopsies stained with anti p53 monoclonal antibody (DO-7), and graded the degree of staining (0, +, ++, +++). Samples from a cohort of 90 patients with CLL were studied (median age 60 years, range 30-89 years; 57 patients were (63%) previously untreated, 22 patients (24%) had received one or two prior regimens, 11 patients had received (12%) three to seven regimens. The overall percentage of cells positive for p53 staining was a median of 43 (range 1-88). No investigator effect was detected either in overall percentage cells rated p53 positive or on the degree of staining (Pearson's correlation coefficient 0.980, P-value < 0.001). A Cox proportional hazards model showed that the percentage of ++ and +++ p53-positive cells correlated with various prognostic factors in CLL (P < 0.0001). A multivariate model incorporating prior therapy, Rai stage, beta2 microglobulin (beta2M) and p53 expression showed that only the percentage of p53-positive cells and beta2M were predictive of survival, and enabled the development of a highly predictive model of survival based on these two parameters. PMID- 12752097 TI - Cytogenetic abnormalities in childhood acute myeloid leukaemia: a Nordic series comprising all children enrolled in the NOPHO-93-AML trial between 1993 and 2001. AB - Between 1993 and 2001, 318 children were diagnosed with acute myeloid leukaemia (AML) in the Nordic countries. The patient group comprised 237 children < 15 years of age with de novo AML, 42 children < 15 years with Down syndrome (DS) and de novo AML, 18 adolescents 15-18 years of age with de novo AML, and 21 children < 15 years with treatment-related AML (t-AML). The first group was all-inclusive, yielding an annual childhood de novo AML incidence of 0.7/100 000. Cytogenetic analyses were successful in 288 cases (91%), and clonal chromosomal abnormalities were detected in 211 (73%). The distribution of ploidy levels were pseudodiploidy (55%), hyperdiploidy (34%) and hypodiploidy (11%). The most common aberrations (> 2%) were + 8 (23%) (as a sole change in 6.2%), 11q23-translocations, including cryptic MLL rearrangements (22%) [t(9;11)(p21-22;q23) in 11%], t(8;21)(q22;q22) (9.0%), inv(16)(p13q22) (6.2%), -7/7q- (5.2%), and t(15;17)(q22;q12) (3.8%). Except for +8, these abnormalities were rare in group 2; only one DS patient had a t(8;21) and none had 11q23-translocations, t(15;17) or inv(16). In the t-AML group, three cases displayed 11q23-rearrangements, all t(9;11); and there were no t(8;21), t(15;17) or inv(16). Overall, the observed frequencies of t(8;21) and t(15;17) were lower, and frequencies of trisomy 8 and 11q23-translocations higher, than in previous studies. Furthermore, seven abnormalities that were previously reported as only single AML cases were also seen, meaning that der(4)t(4;11)(q26-27;q23), der(6)t(1;6)(q24-25;q27), der(7)t(7;11)(p22;q13), inv(8)(p23q11-12), t(11;17)(p15;q21), der(16)t(10;16)(q22;p13) and der(22)t(1;22)(q21;q13) are now classified as recurrent abnormalities in AML. In addition, 37 novel aberrations were observed, 11 of which were sole anomalies. PMID- 12752099 TI - Localized non-Hodgkin's lymphoma with B-cell histology: cure without cyclophosphamide? A report of the United Kingdom Children's Cancer Study Group on studies NHL 8501 and NHL 9001 (1985-1996). AB - We have examined the outcome for children treated on two consecutive United Kingdom Children's Cancer Study Group studies of localized B-cell non-Hodgkin's lymphoma (NHL). The first study (NHL 8501; 1985-1989) included cyclophosphamide in the treatment regimen at a total cumulative dose of 4 g/m2 whereas the regimen in the succeeding study (NHL 9001; 1990-1996) did not include cyclophosphamide. Ninety children with confirmed B-cell NHL were treated in the two studies (NHL 8501, n = 33 and NHL9001, n = 57). With a median follow-up of 7.5 years, overall survival for localized B-cell NHL did not differ between the two regimens with observed 3-year survivals of 94%[95% confidence interval (CI) 80-98%] and 89% (95% CI 79-95%) respectively (P = 0.47). There was also no difference in the event-free survival between children treated on regimen NHL 8501 and NHL 9001 [91% (95% CI 76-97%) vs 84% (95% CI 73-92%) after 3 years; P = 0.34]. Although the difference in the number of failed remissions between NHL 8501 and 9001 (0/33 vs 6/57) approached statistical significance (P = 0.08, Fisher's exact test), there was no overall statistical difference between the treatment failures on either regimen (P = 0.34). Substantial long-term survival can be achieved for many children with localized B-cell NHL without the use of cyclophosphamide. Further studies are needed to identify whether all clinical or histopathological subgroups will benefit equally from the omission of cyclophosphamide. PMID- 12752100 TI - Recombinant humanized anti-CD40 monoclonal antibody triggers autologous antibody dependent cell-mediated cytotoxicity against multiple myeloma cells. AB - Multiple myeloma (MM) is currently incurable, and novel therapies are needed. In this study, we examined a novel recombinant humanized monoclonal antibody against CD40 (rhuCD40 mAb) and demonstrate for the first time that rhuCD40 mAb induces antibody-dependent cell-mediated cytotoxicity (ADCC) against CD40-positive MM cells. Importantly, we show that rhuCD40 mAb induces autologous ADCC against primary patient MM cells, without triggering ADCC against normal B cells. This study, therefore, both demonstrates that rhuCD40 mAb triggers autologous ADCC against patient MM cells and provides the framework for the clinical evaluation of rhuCD40 mAb immunotherapy to improve patient outcome in MM. PMID- 12752102 TI - Immunoplatelet counting: potential for reducing the use of platelet transfusions through more accurate platelet counting. AB - Research is required to determine the optimal approach for prophylactic platelet transfusions in patients with haematological malignant disorders. It has been suggested that thresholds for prophylactic platelet transfusions of platelet counts below 10 x 109/l should be investigated, as these may be equivalent in clinical effectiveness and associated with lower costs and fewer complications. An important concern in such investigation is the accurate estimation of platelet counts below 10 x 109/l. This study aimed to further examine the potential reduction in platelet usage that could be made if a lowered platelet transfusion threshold of 5 x 109/l was used in conjunction with an immunoplatelet counting method. Clinical and laboratory data from 130 haematology patients were used. Standard platelet counting was performed using Bayer H3 and ABX Argos analysers. Immunoplatelet counting was performed by flow cytometry using anti-CD61. The potential for reducing platelet transfusions included consideration of clinical criteria that influence prophylactic platelet transfusion use. The results indicated that the use of an immunoplatelet count with a 5 x 109/l platelet transfusion threshold would potentially reduce the number of transfusions by 10.4% in comparison with a 10 x 109/l threshold and standard automated platelet counting with the ABX Argos analyser, and increase the number of transfusions by 5.4% in comparison with the same threshold using the Bayer H3 analyser. The immunoplatelet count may aid the clinical decision to transfuse platelets, but would not necessarily lead to a reduced use of platelet transfusions. PMID- 12752101 TI - Interleukin-1 genotype and outcome of unrelated donor bone marrow transplantation. AB - The interleukin 1 (IL-1) gene family includes three members (IL-1-alpha, IL-1 beta and IL-1Ra) that mediate immune and inflammatory responses through two specific cell surface receptors. Cytosine to thymine transitions at codons -889 and -511 in the IL-1-alpha and IL-1-beta genes, respectively, and an 86-base pair repeat in the IL-1Ra are believed to influence gene transcription. We have genotyped these three polymorphisms in 90 donor/recipient pairs undergoing unrelated donor bone marrow transplantation (BMT) at the University of Minnesota. We found no association between the occurrence of acute GVHD and donor and/or recipient polymorphisms of any of the three IL-1 genes. The presence of at least one IL-1alpha- 889 T allele in the donor was associated with significantly improved survival in univariate analysis (survival at 1 year 40% C/C donor, 68% T/C donor, 75% T/T donor, P < 0.01). Multiple regression analysis showed that if the donor and recipient each possessed the IL-1alpha T allele there was significantly improved survival [relative risk (RR) 0.2, P < 0.01] and decreased treatment-related mortality (TRM; RR 0.2, P = 0.01). The presence of the IL-1beta T allele in donor and recipient was also associated with improved survival (RR 0.2, P < 0.01) and decreased TRM (RR 0.1, P < 0.01). These data suggest that donor polymorphism in IL-1alpha and IL-1beta might influence survival after unrelated donor BMT, but does not alter risk of GVHD. PMID- 12752103 TI - Recombinant human macrophage colony-stimulating factor-induced thrombocytopenia in dogs. AB - To characterize recombinant human macrophage-colony stimulating factor (rhM-CSF) associated thrombocytopenia (TCP), in vivo studies were performed in dogs, including the biodistributions and recoveries of radiolabelled autologous and allogeneic platelets. rhM-CSF induced a reversible, dose-dependent decrease in platelet counts. The number of megakaryocytes in spleen and marrow of rhM-CSF treated dogs was increased two to threefold. Recoveries of allogeneic platelets transfused from rhM-CSF-treated donors into tolerized recipients (n = 3) were not significantly different from allogeneic baseline studies (93 +/- 10% of baseline values at 24 h and 90 +/- 1% at 40 h), whereas autologous platelets infused back into rhM-CSF-treated donors had decreased recoveries (45 +/- 2% of baseline values at 24 h, P = 0.03 and 20 +/- 4% at 40 h, P = 0.001). Platelet biodistribution studies showed increased accumulation of radiolabelled platelets over the spleens and livers of rhM-CSF-treated dogs. Histochemistry showed increased levels of platelet-specific antigen (CD41; glycoprotein IIb) associated with Kupffer cells. The sensitivity of platelets from rhM-CSF-treated dogs to activation from thrombin, as measured by expression of P-selectin (CD62P), was not significantly different when compared with baseline studies (P = 0.18; n = 4). These results support the concept that rhM-CSF induces an activation of the monocyte-macrophage system (MMS), which causes a reversible TCP in a dog model. PMID- 12752104 TI - The role of P-selectin in the immune destruction of platelets. AB - Antibody-mediated platelet destruction is a poorly understood process, although several lines of evidence suggest that Fcgamma receptor (FcgammaR)-expressing splenic macrophages may be involved. In this study, chemiluminescence (CL) was used to measure the in vitro metabolic response of human monocytes to platelets sensitized with a human immunoglobulin (Ig)G1 recombinant antihuman platelet antigen-1a (anti-HPA-1a) antibody (B2G1; P-hrIgG1). CL responses were inhibited, but not abrogated, in the presence of 10 micro g/ml human IgG or murine IgG2a, suggesting that FcgammaRI was principally involved. Experiments to determine the effect of Fab fragments to FcgammaRII found that CL responses to P-hrIgG1 were significantly enhanced, indicating that crosslinking of monocyte FcgammaRII by platelet-bound hIgG may modulate concomitant activation by FcgammaRI. Several observations suggested that the CL responses to P-IgG were dependent on the activation of resting platelets during their co-culture with monocytes and their subsequent P-selectin-mediated adhesion. First, the magnitude of the CL response was related to the level of P-selectin expression following platelet activation with alpha-thrombin. Second, CL responses were inhibited in the presence of antibodies that block the binding of P-selectin to P-selectin glycoprotein ligand 1 but not when platelets were pretreated and then washed. Third, the addition of anti-HPA-1a to monocytes from HPA-1a-negative donors preincubated with HPA-1a positive platelets resulted in rapid CL responses. Finally, PGI2 inhibited the CL response to resting P-hrIgG1. Thus, evidence is presented that the interaction of human monocytes with P-hrIgG1 is mediated by FcgammaRI, modulated via FcgammaRII, and enhanced by the presence of P-selectin on the platelet membrane. PMID- 12752105 TI - Asymptomatic carriership of factor V Leiden and genotypes of the fibrinogen gene cluster. AB - We investigated the role of frequent fibrinogen polymorphisms in venous thromboembolic disease in conjunction with inherited thrombophilia. Two hundred unrelated subjects, all carriers of the factor V R506Q mutation (FV Leiden), were genotyped at the fibrinogen gene cluster. Among these subjects, 100 had experienced previous venous thromboembolism (VTE) and 100 were still asymptomatic for VTE. Significant differences were observed between the groups for the BclI polymorphism (P = 0.004). Scanning, by sequencing the DNA regions flanking the BclI marker, revealed new polymorphisms, a C to T transition and a G to T transversion at 1520 and 3369 base pairs 3' to the beta gene stop codon respectively. These markers showed less association with the clinical phenotype than BclI itself. A combined genotype including 10 markers was more frequent among the asymptomatic subjects (17%) than among patients (3%), and was associated with a reduction in fibrinogen antigen level (2.42 +/- 0.35 vs 2.69 +/ 0.41 g/l, P = 0.028) among the asymptomatic subjects. Our data suggest that, in the presence of inherited thrombophilia, frequent fibrinogen polymorphisms may interact to modulate the risk of venous thromboembolism. PMID- 12752106 TI - Lupus anticoagulant associated with transient severe factor X deficiency: a report of two patients presenting with major bleeding complications. AB - Acquired factor X (FX) deficiency is rare, but has been reported in diverse disease states, including systemic amyloidosis and respiratory infections. FX deficiency associated with lupus anticoagulant (LA) and a bleeding diathesis has not been previously reported. We report two patients both of whom presented with a severe bleeding diathesis after a preceding respiratory infection due to isolated FX deficiency associated with a LA. The FX deficiency and LA were transient. We conclude that patients with LA may rarely present with severe acquired FX deficiency. This may be another mechanism whereby patients with antiphospholipid antibodies present with bleeding complications. PMID- 12752107 TI - Progenitor cells divide symmetrically to generate new colony-forming cells and clonal heterogeneity. AB - Self-renewal is the most fundamental property of haemopoietic stem and progenitor cells. However, because of the need to produce differentiated cells, not all cell divisions involve self-renewal. We have used a colony replating assay to follow the fates of individual haemopoietic progenitor cell clones. For this, human myeloid colony-forming cells (CFCs) were cultured by standard methodology. Onset of proliferation and growth rates were established by a video recording method. Individual colonies were replated several times to document the rate of clonal extinction, and the numbers of secondary, tertiary and quaternary CFCs. The clonogenic population exhibited similar kinetics in terms of onset of proliferation and growth rate. Clonal extinction was progressive so that only 30 +/- 7% (mean +/- standard error of the mean; n = 4) of the original primary colonies formed quaternary colonies after the third replating step. However, individual primary CFCs that produced colonies throughout the experiment generated, on average, 40 +/- 8 secondary and tertiary CFCs overall. The values obtained in standard culture conditions were modified when granulocyte colony stimulating factor (G-CSF) or G-CSF plus interleukin 3 were used to stimulate colony growth, showing that the kinetics of colony formation respond to extrinsic regulation. Examination of the replating potential of individual secondary colonies in the clones demonstrated that they generated different numbers of tertiary colonies. The data best fit a stochastic model of haemopoietic cell development where event probabilities can be modified by extracellular factors. PMID- 12752108 TI - Human bone marrow endothelial cells elaborate non-stromal-cell-derived factor-1 (SDF-1)-dependent chemoattraction and SDF-1-dependent transmigration of haematopoietic progenitors. AB - This study investigated human bone marrow endothelial cells (BMEC) chemoattractive activity in relation to haematopoietic cell trafficking. BMEC conditioned medium induced chemoattraction of haematopoietic progenitor cells. Migration was not inhibited by pretreating the cells with pertussis toxin (PTX) or 12G5, indicating that the chemoattractive activity was not dependent on stromal-cell-derived factor-1 (SDF-1). Spontaneous migration, but not SDF-1 mediated chemotaxis of haematopoietic progenitors, was better supported by BMEC as compared with umbilical vein endothelial cells. The superior migration was abolished by pretreating the cells with PTX, indicating that BMEC-derived SDF-1 favours bone marrow endothelium, with better transmigration of haematopoietic progenitors. PMID- 12752109 TI - Endotoxaemia modulates Toll-like receptors on leucocytes in humans. AB - The modulation of Toll-like receptors (TLR) 1, 2 and 4 was studied during experimental human endotoxaemia. Healthy volunteers received 2 ng/kg of lipopolysaccharide (LPS) endotoxin (n = 10). TLR1, 2 and 4 expression occurred on monocytes and neutrophils, with monocytes expressing higher baseline levels of TLR2. LPS infusion downmodulated TLR4 expression on neutrophils, with maximal downregulation occurring at 24 h (-62% from baseline; P < 0.03 versus baseline). Monocyte TLRs were upregulated in vivo (TLR1 and 2), and in vitro (TLR1, 2 and 4) 8 h after LPS bolus (P < 0.05 versus baseline). Therefore, neutrophils and monocytes differentially express surface TLRs, and endotoxaemia differentially regulates TLR expression. PMID- 12752111 TI - The molecular basis for the thalassaemias in Sri Lanka. AB - The beta-globin gene mutations and the alpha-globin genes of 620 patients with the phenotype of severe to moderate thalassaemia from seven centres in Sri Lanka were analysed. Twenty-four beta-globin gene mutations were identified, three accounting for 84.5% of the 1240 alleles studied: IVSI-5 (G-->C) 56.2%; IVSI-1 (G ->A) 15.2%; and haemoglobin E (codon (CD)26 GAG-->GAA) 13.1%. Three new mutations were found; a 13-bp deletion removing the last nucleotide in CD6 to CD10 inclusively, IVSI-129 (A-->C) in the consensus splice site, and a frame shift, CD55 (-A). The allele frequency of alpha+ thalassaemia was 6.5% and 1.1% for alpha3.7 and -alpha4.2 deletions respectively. Non-deletion alpha-thalassaemia was not observed. Triplicate or quadruplicate alpha-globin genes were unusually common. In 1.5% of cases it was impossible to identify beta-thalassaemia alleles, but in Kurunegala detailed family studies led to an explanation for the severe thalassaemia phenotype in every case, including a previously unreported instance of homozygosity for a quadruplicated alpha-globin gene together with beta thalassaemia trait. These findings have implications for the control of thalassaemia in high-frequency populations with complex ethnic histories. PMID- 12752110 TI - Oxidative stress is involved in hydroxyurea-induced erythroid differentiation. AB - Hydroxyurea (HU), an inhibitor of DNA synthesis, can also induce haemoglobinization in certain erythroid cell lines. In this study, we report that intracellular peroxides levels were increased in HU-treated murine erythroleukaemia (MEL) cells and that l-acetyl-N-cysteine (LNAC), a potent reducing reagent, had a significant inhibitory effect on the HU-mediated induction of beta-globin, delta-aminolaevulinate synthase mRNA expression and haemoglobinization of MEL cells. In contrast, the addition of LNAC to dimethyl sulphoxide (DMSO)-treated MEL cells had a much smaller effect on the number of haemoglobinized cells. These findings suggest that oxidative stress is involved in HU-mediated induction of erythroid differentiation and that HU induces MEL cell differentiation by a mechanism different to that involved in DMSO-mediated differentiation. Our findings also suggest that the induction of MEL cell differentiation by HU does not involve RAS-MAP (mitogen-activated protein) kinase signalling. PMID- 12752112 TI - Cross-resistance of imatinib mesylate and 17-AAG in imatinib-resistant cells that overexpress BCR-ABL. PMID- 12752113 TI - The expression of prion protein (PrPc) by endothelial cells: an in vitro culture induced artefactual phenomenon? PMID- 12752114 TI - The G277S mutation in transferrin does not disturb function. PMID- 12752115 TI - Hereditary haemolytic anaemias and parvovirus infections in Jehovah's Witnesses. PMID- 12752117 TI - The British Association of Dermatologists therapeutic guidelines: can we AGREE? PMID- 12752118 TI - Ultraviolet A1 phototherapy. AB - Long-wavelength ultraviolet A (340-400 nm; UVA1) therapy is currently available in only a few dermatology departments. Equipment capable of delivering this waveband has been available since 1981, but it is only over the past decade that increasing numbers of studies assessing the potential of this as a therapy have been published. High-dose UVA1, which requires expensive and space-occupying apparatus, is effective as a monotherapy for acute flares of atopic dermatitis, but it has not yet been formally assessed as an adjunct, rather than as an alternative to conventional therapies including potent and very potent topical corticosteroids. Low-dose (which can be administered using a standard phototherapy cubicle fitted with appropriate lamps) and medium-dose UVA1 may be less effective for this indication. Another condition for which UVA1 is effective, and is particularly promising because we have no reliably effective treatment already, is localized scleroderma. It also appears to be effective in systemic lupus erythematosus (although it is not yet clear when it is indicated, and its safety needs to be assessed in more patients) and in polymorphic light eruption (although there have been no studies suggesting that UVA1 will have any advantages over standard prophylactic phototherapies). Open studies and case series suggest that UVA1 may prove beneficial for various other diseases, including cutaneous T-cell lymphoma, lichen sclerosus, keloids, systemic sclerosis and hand dermatitis. In the centres where it is available, UVA1 has already proved a useful addition to the range of phototherapies previously available. However, much more research is needed to confirm its efficacy for many of its potential indications, and to determine when and how it should be used. PMID- 12752119 TI - Mutational analysis of CDKN2A genes in patients with squamous cell carcinoma of the skin. AB - BACKGROUND: Nonmelanoma skin cancers [squamous cell carcinomas (SCC) and basal cell carcinomas (BCC)] are the most common neoplasias of the Caucasian population. OBJECTIVES: The purpose of our study was to determine the involvement of CDKN2A genes in the development of sporadic nonmelanoma skin cancer in Greek patients. PATIENTS AND METHODS: Allelic imbalance analysis was performed in 22 SCC and five Bowen's disease specimens. Mutational analysis was performed on exons 1alpha, 1beta and 2 of the CDKN2A locus in 22 SCC, five Bowen's disease and 39 BCC specimens. Exon 1alpha was additionally screened in 28 BCC specimens to complete the mutational analysis of a previous study. RESULTS: Overall, 52% (14 of 27) of the SCC and Bowen's disease specimens exhibited loss of heterozygosity (LOH) in at least one microsatellite marker, whereas, only two of 27 (7%) exhibited microsatellite instability. LOH in 9p appears to be equally involved in both BCC and SCC tumours. Exons 1alpha, 1beta and 2 of the CDKN2A locus were screened for mutations. A Val28Gly substitution in exon 1alpha and a CCC-->TTT (Ala57Val and Arg58Ter) substitution in exon 2, resulting in a change in the amino acid sequence, are reported for the first time in two SCCs, the latter being indicative of a combination of an ultraviolet (UV) radiation-induced mutation and a point mutation. A previously described polymorphism of CDKN2A, the gene for p16INK4a, Ala148Thr, was also detected in an allelic frequency of 3.72%. No mutation was found in any of the five Bowen's disease specimens, or in exon 1beta of CDKN2A, also the gene for p14ARF. CONCLUSIONS: Mutations and the high incidence of 9p LOH detected in our SCC samples imply that inactivation of CDKN2A genes, via allelic loss and/or mutation (probably UV-induced) may play a significant role in nonmelanoma skin cancer development, particularly in the more aggressive SCC type. PMID- 12752120 TI - Novel mutations in GJB2 encoding connexin-26 in Japanese patients with keratitis ichthyosis-deafness syndrome. AB - BACKGROUND: Germline missense mutations in the GJB2 gene that encodes connexin-26 (Cx26) have recently been found to be the cause of the keratitis-ichthyosis deafness (KID) syndrome. OBJECTIVES: To define the GJB2 mutations in three Japanese patients with KID syndrome. METHODS: Genomic DNA was extracted from peripheral blood and used to amplify the GJB2 gene. Direct sequencing and endonuclease digestion were used for mutation analysis and DNA-based diagnosis. RESULTS: We identified two heterozygous mis-sense mutations (D50Y, D50N) in the GJB2 gene in three Japanese patients with KID syndrome. All mutations were located on the first extracellular domain of Cx26. CONCLUSIONS: These data expand the GJB2 mutation database and show that a dominant mutation of Cx26 can cause KID syndrome in Japanese patients. PMID- 12752121 TI - Identification of novel genes for secreted and membrane-anchored proteins in human keratinocytes. AB - BACKGROUND: Both intercellular and intracellular signals are transduced primarily by interactions of secreted and/or membrane-anchored polypeptides, and they play a pivotal role in regulating proliferation, differentiation and apoptosis of keratinocytes within the epidermis. Despite recent identification of these polypeptides, it is likely that several important molecules remain undisclosed. OBJECTIVES: To identify novel genes encoding secreted or membrane-anchored polypeptides expressed by human keratinocytes. METHODS: We employed a signal sequence (SS) trap of a 5'-end-enriched cDNA library prepared from primary cultured human keratinocytes. Gene expression analysis was performed using Northern blotting. R Screening of 4018 cDNA clones yielded 82 positive clones (57 independent genes), most of which encoded SSs in their N-termini. Most of the positive clones were known genes registered in the GenBank database. Seven genes were identified in the EST database, four of which encoded novel membrane anchored polypeptides with features of type I transmembrane proteins; the other three genes encoded novel non-type I transmembrane polypeptides. These EST genes were expressed differentially by keratinocytes subjected to low vs. high calcium concentrations and by basal vs. squamous cell carcinomas. CONCLUSIONS: Using the SS trap, we isolated many genes known to be involved in constituting epidermal structures and others that had not previously been associated with keratinocytes. In addition, we identified novel genes (EST genes) that differ in kinetics of gene expression in keratinocyte differentiation. Our results validate the effective use of this SS trap method for identifying secreted and membrane anchored polypeptides expressed by human keratinocytes. The identification will better illuminate the molecular mechanisms responsible for co-ordinated regulation of epidermal homeostasis. PMID- 12752122 TI - Association of SPINK5 gene polymorphisms with atopic dermatitis in the Japanese population. AB - BACKGROUND: Netherton's syndrome (NS) is an autosomal recessive disorder characterized by trichorrhexis invaginata ('bamboo hair'), congenital ichthyosiform erythroderma and an atopic diathesis. NS has recently been shown to be due to a defect in the SPINK5 gene, encoding LEKTI, a 15-domain serine protease inhibitor. SPINK5 maps to chromosome 5q31-q32, and has been suggested to be a locus predisposing to atopy in general. Recently, coding polymorphisms in SPINK5 exons 13 and 14 have been reported to be associated with atopy, asthma and atopic dermatitis (AD). OBJECTIVES: To examine whether these polymorphisms are also associated with AD in Japan. METHODS: We characterized eight polymorphisms in SPINK5 exons 13 and 14 in 124 Japanese patients with AD and 110 healthy controls. The polymorphisms we examined were IVS12-26C-->T, IVS12-10A-->G, 1103A- >G (Asn368Ser, in exon 13), 1156G-->A (Asp386Asn, in exon 13), 1188T-->C (His396His, in exon 13), IVS13-50G-->A, 1258G-->A (Glu420Lys, in exon 14) and IVS14+19G-->A. RESULTS: We found significant associations between seven of these polymorphisms and AD in Japanese patients. CONCLUSIONS: This study confirms the previous suggestion of an association between SPINK5 and AD. PMID- 12752123 TI - Normal keratinocytes express Toll-like receptors (TLRs) 1, 2 and 5: modulation of TLR expression in chronic plaque psoriasis. AB - BACKGROUND: Toll-like receptors (TLRs) are part of the innate immune system involved in the response to microbial pathogens. TLR2 recognizes various ligands expressed by Gram-positive bacteria, while TLR3, TLR4 and TLR5 are specific for double-stranded RNA, Gram-negative lipopolysaccharides and bacterial flagellin, respectively. OBJECTIVES: To determine, firstly, whether epidermal keratinocytes of normal skin express TLRs and, secondly, whether modulation of TLR expression occurs in psoriasis, an inflammatory skin disease associated with certain microorganisms such as streptococci, staphylococci and yeasts. METHODS: Eight samples of normal, and 15 samples of lesional and nonlesional psoriatic skin were stained with polyclonal antibodies specific for TLR1-5 using an avidin-biotin peroxidase technique. RESULTS: Epidermal keratinocytes in normal skin constitutively expressed TLR1, TLR2 and TLR5, while TLR3 and TLR4 were, in most cases, barely detectable. Cytoplasmic TLR1 and TLR2 were expressed throughout the epidermis, with higher staining of the latter on basal keratinocytes, while TLR5 expression was concentrated in the basal layer. In contrast, in lesional epidermis from patients with psoriasis, TLR2 was more highly expressed on the keratinocytes of the upper epidermis than on the basal layer, while TLR5 was downregulated in basal keratinocytes compared with corresponding nonlesional psoriatic epidermis. In addition, nuclear TLR1 staining was observed in the upper layers of both nonlesional and lesional psoriatic epidermis, but not in that of normal skin. CONCLUSIONS: These findings suggest that TLRs expressed by epidermal keratinocytes constitute part of the innate immune system of the skin. The relevance of altered keratinocyte TLR expression in psoriasis remains to be determined. PMID- 12752124 TI - Expression of NF-kappaB in epidermis and the relationship between NF-kappaB activation and inhibition of keratinocyte growth. AB - BACKGROUND: Nuclear factor-kappaB (NF-kappaB) is a transcription factor involved in a number of signalling pathways in many cell types. NF-kappaB in mice has been implicated as an important regulator of keratinocyte proliferation and differentiation. OBJECTIVES: To evaluate the role of NF-kappaB in keratinocyte growth in human beings, we examined its expression in keratinocytes both in culture and in situ, and studied the relationship between NF-kappaB activation and the inhibition of keratinocyte proliferation induced by known modulators of keratinocyte growth. METHODS: The expression of subunits of the NF-kappaB family was examined in human skin, primary cultured keratinocytes and an immortalized keratinocyte line by immunohistochemistry and reverse transcriptase-polymerase chain reaction analysis. NF-kB activation was examined in keratinocytes treated with various modulating agents by electrophoretic mobility shift assay (for DNA binding activity) and by immunocytochemistry (nuclear translocation). The proliferative capacity of treated keratinocytes was also examined by 3H-thymidine incorporation, cell cycle analysis, and expression of Ki-67, a nuclear marker for cell proliferation. The involvement of NF-kappaB was assessed using sodium salicylate, which inhibits NF-kappaB activation. RESULTS: The NF-kappaB subunits, p50, p65, RelB, and c-Rel (but not p52), were detected in keratinocytes and in normal epidermis at mRNA and protein levels. The four subunits were expressed in a cytoplasmic (rather than a nuclear) pattern in both basal and suprabasal keratinocytes. Phorbol myristate acetate (PMA), tumour necrosis factor alpha, and interferon gamma each activated NF-kappaB and inhibited keratinocyte proliferation. Lipopolysaccharide and dexamethasone did not activate NF-kappaB and had the least effect on proliferation. Finally, a high concentration of calcium (Ca2+) and retinoic acid each failed to activate NF-kappaB, but were potent inhibitors of keratinocyte proliferation, respectively. PMA-induced cell cycle arrest of keratinocytes was blocked by pretreatment with sodium salicylate. CONCLUSIONS: NF-kappaB is constitutively expressed in a resting state in both human cultured keratinocytes and the epidermis. Activation of NF-kappaB is required for PMA-induced keratinocyte growth arrest. PMID- 12752125 TI - The mechanism of epidermal hyperpigmentation in cafe-au-lait macules of neurofibromatosis type 1 (von Recklinghausen's disease) may be associated with dermal fibroblast-derived stem cell factor and hepatocyte growth factor. AB - BACKGROUND: The mechanism of the accentuated melanization in cafe-au-lait macules (CALMs) in patients with neurofibromatosis type 1 (NF1; von Recklinghausen's disease) has not been elucidated. OBJECTIVES: To clarify the mechanism involved in the hyperpigmentation of CALMs in NF1. METHODS: Using enzyme-linked immunosorbent assay (ELISA) and reverse transcriptase-polymerase chain reaction (RT-PCR) analysis of cultured cells, we measured the levels of cytokines produced and secreted by keratinocytes and fibroblasts derived from CALMs (group RC: Recklinghausen CALM) skin, compared with cells derived from the skin of normal individuals (group NN: Normal skin of Normal individuals) and cells derived from non-CALM skin of NF1 patients (group RN: Recklinghausen Non-CALM). RESULTS: ELISA revealed that the secretion of hepatocyte growth factor (HGF) and stem cell factor (SCF) by cultured fibroblasts was significantly elevated in group RC compared with groups RN and NN. In parallel, semiquantitative real-time RT-PCR of HGF and SCF mRNAs demonstrated increased expression of both types of transcripts by cultured fibroblasts in group RC compared with group NN. In contrast, the secretion of endothelin-1 and granulocyte/macrophage colony-stimulating factor by cultured keratinocytes occurred at a similar level among all three groups, RC, RN and NN. CONCLUSIONS: These findings suggest that increased secretion of HGF and SCF by dermal fibroblasts may be associated with the accentuated epidermal melanization observed in CALMs in the skin of NF1 patients. PMID- 12752126 TI - Diagnosis of pigmented skin lesions by dermoscopy: web-based training improves diagnostic performance of non-experts. AB - BACKGROUND: Dermoscopy has been shown to enhance the diagnosis of melanoma. However, use of dermoscopy requires training and expertise to be effective. OBJECTIVES: To determine whether an Internet-based course is a suitable tool in teaching dermoscopy, and to evaluate the diagnostic value of pattern analysis and diagnostic algorithms in colleagues not yet familiar with this technique. METHODS: Sixteen colleagues who were not experts in dermoscopy were asked to evaluate the dermoscopic images of 20 pigmented skin lesions using different diagnostic methods (i.e. pattern analysis, ABCD rule, seven-point checklist and Menzies' method), before and after an Internet-based training course on dermoscopy. Mean +/- SEM sensitivity, specificity and diagnostic accuracy, and kappa (kappa) intraobserver agreement were evaluated for each diagnostic method before and after training for the 16 participants. Differences between mean values were assessed by means of two-tailed Wilcoxon rank-sum tests. RESULTS: There was a considerable improvement in the dermoscopic melanoma diagnosis after the Web-based training vs. before. Improvements in sensitivity and diagnostic accuracy were significant for the ABCD rule and Menzies' method. Improvements in sensitivity were also significant for pattern analysis, whereas the sensitivity values were high for the seven-point checklist in evaluations both before and after training. No significant difference was found for specificity before and after training for any method. There was a significant improvement in the kappa intraobserver agreement after training for pattern analysis and the ABCD rule. For the seven-point checklist and Menzies' method there was already good agreement before training, with no significant improvement after training. CONCLUSIONS: We demonstrated that Web-based training is an effective tool for teaching dermoscopy. PMID- 12752127 TI - Local recurrence in melanoma in situ: influence of sex, age, site of involvement and therapeutic modalities. AB - BACKGROUND: Melanoma in situ (MIS) occurs on various body sites, in various age groups, and is managed by a variety of treatment modalities. Despite early treatment, recurrences may be encountered. OBJECTIVES: To evaluate the influence of sex, age, body site and treatment modalities on recurrence rate in MIS. Methods Histologically confirmed cases of MIS from our dermatopathological database (n = 1351) from 1990 to 2000 were statistically analysed with respect to epidemiological characteristics, treatment modalities and outcome. Treatment modalities of the included MIS were evaluated by searching for data in the medical records and histopathological data sheets. RESULTS: There was a predominance of female patients (60.8%), and of involvement of the head and neck (53.4%). Total excision was performed in 95.9% of all patients; the remainder received cryotherapy, laser therapy or radiotherapy. In 265 patients, no data on definitive treatment were available. Alternatives to total excision were particularly performed in patients with advanced age and with lesions localized on the face. The mean +/- SD 5-year recurrence rate was 6.8 +/- 1.3% for surgically removed lesions, but was 31.3 +/- 8.5% for lesions treated by other modalities (log rank test: P < 0.0001). In a multivariate approach, mode of therapy and site of involvement, but not age, were significant prognostic variables (Cox proportional hazard model: P < 0.01). CONCLUSIONS: In MIS, treatment modalities other than surgical excision may be used in certain situations, but carry a significantly increased risk of local recurrence. PMID- 12752128 TI - The relevance of peripheral blood T-helper 1 and 2 cytokine pattern in the evaluation of patients with mycosis fungoides and Sezary syndrome. AB - BACKGROUND: There is evidence that a T-helper (Th) 2 cytokine pattern dominates in the peripheral blood as well as in tissue of patients with Sezary syndrome (SS), and that the malignant clone is of Th2 phenotype. However, there are conflicting studies on the cytokine pattern in the peripheral blood in different stages of cutaneous T-cell lymphoma (CTCL). OBJECTIVES: To examine, by means of flow cytometry (FC), the Th1/Th2 cytokine profile [cytoplasmic interferon (IFN) gamma/interleukin (IL)-4] in peripheral blood T cells from patients with mycosis fungoides (MF) and SS, the most common forms of CTCL, and to correlate their expression with clinical stage, clonality and T-cell immunophenotype changes in order to evaluate their relevance in CTCL progression. METHODS: We investigated by FC the percentage of CD3+ T cells expressing cytoplasmic IFN-gamma and IL-4 after stimulation in blood specimens of 43 CTCL patients (32 stage I-II and 11 stage III-IV), eight of whom were erythrodermic. Next, we compared cytoplasmic IFN-gamma and IL-4 expression between patients of different stages and controls, and correlated our findings to T-cell receptor (TCR)-gamma gene rearrangement, used as a marker of clonality, and changes in T-cell immunophenotype (CD4+, CD8+, CD4+/CD7-, CD4+/CD25+) and natural killer cells. Polymerase chain reaction amplification of the TCR-gamma gene was performed in 41 blood and 26 skin specimens. We also examined the cytokine expression pattern in patients with erythrodermic MF and SS. RESULTS: A significantly higher frequency of CD3+/IL-4+ T cells was found in late (III-IV) compared with early (I-II) CTCL patients (P = 0.002) or controls (P < 0.001). There were significant positive correlations between the percentages of CD3+/IL-4+ and the percentages of CD3+/CD4+ T cells (r = 0.385, P = 0.05), CD4+/CD7- T cells (r = 0.335, P < 0.05) and CD4+/CD25+ T cells (r = 0.433, P = 0.01); there was a negative correlation between the percentages of CD3+/IL-4+ and CD3+/CD8+ T cells (r = -0.463, P = 0.005) and a positive correlation between the percentages of CD3+/IFN-gamma+ and CD3+/CD8+ T cells (r = 0.368, P = 0.02). Increased percentages of CD3+/IL-4+, CD3+/CD4+ and CD4+/CD7- T lymphocytes were associated with the presence of clonality (P = 0.025, P < 0.001 and P = 0.0031, respectively). All independent variables showed a statistically significant difference between SS and erythrodermic MF patients, or controls, apart from cytoplasmic IL-4, which was high both in erythrodermic MF and SS patients compared with controls (P = 0.003 and P = 0.008, respectively). In multiple regression logistic analysis, the probability of belonging to advanced CTCL stages was associated only with increased cytoplasmic IL-4 (P = 0.007, odds ratio 1.13, 95% confidence interval 1.033-1.229). CONCLUSIONS: Increased T-cell cytoplasmic IL-4 is more frequent in late CTCL stages, correlates with T-cell immunophenotype changes found in advanced disease and is associated with clonality. Increased cytoplasmic IL-4 is frequent both in erythrodermic MF and SS patients, in contrast to other variables found increased only in SS, suggesting that IL-4 may be an early indicator of disease progression. Moreover, our results show that increased cytoplasmic IL-4 is the sole predictor of advanced CTCL disease and confirm the relevance of FC determination of IL-4 in the routine evaluation of CTCL cases. PMID- 12752129 TI - Misdiagnosis of hereditary angio-oedema type 1 and type 2. AB - BACKGROUND: Hereditary angio-oedema is a rare, life-threatening, autosomal dominant condition caused by deficiency (type 1) or dysfunction (type 2) of complement C1 inhibitor. Serological assays to measure C1 inhibitor concentration and function are widely available. However, expert interpretation may not be. OBJECTIVE: To review all cases within three NHS Trusts with a putative diagnosis of hereditary angio-oedema. METHOD: Review of laboratory results and clinical notes of 44 cases of presumed hereditary angio-oedema. RESULTS: Audit revealed that 11 of 42 (26%) cases had been incorrectly considered to have a diagnosis of hereditary angio-oedema. Two of 44 had insufficient data to assess. All 11 had low functional C1 inhibitor recorded at presentation. RESULTS: available in these 11 cases at the time of diagnosis showed a normal or borderline C4 level (>or= 50% of mean normal, in contrast to hereditary angio-oedema, where C4 was less than 40% of mean normal) indicating that the low C1 inhibitor levels were a result of sample decay. Cases incorrectly diagnosed were predominantly female and had a mean age at presentation of 40 years (compared with 22 years for type 1 hereditary angio-oedema). Six of the 11 cases were offered C1 inhibitor concentrate (pooled plasma product) as treatment. CONCLUSION: We recommend that all suspected cases of hereditary angio-oedema are reviewed, that specialist advice is sought before making the diagnosis and that the diagnosis is only made after initial abnormal serology is confirmed on a second sample. PMID- 12752130 TI - Thirteen-megahertz ultrasound probe: its role in diagnosing localized scleroderma. AB - BACKGROUND: Ultrasound imaging has been shown to be useful for the evaluation of systemic and localized scleroderma (LS). However, its specificity and sensitivity have not been studied. OBJECTIVES: To define morphological ultrasound diagnostic criteria in LS and to test their sensitivity and specificity with a 13-MHz ultrasound probe. METHODS: Forty plaques in 26 consecutive patients with LS were examined and compared blindly with 17 control plaques in 16 patients with skin diseases where LS was in the differential diagnosis. Data were also compared with a normal control group. Five patients were re-evaluated 12-18 months after the first examination. RESULTS: Ultrasound examination disclosed a characteristic dense image resembling a flattened 'yo-yo'. Undulations of the dermis, disorganization, loss of thickness and thickened hyperechoic bands in the hypodermis, and the 'yo-yo' image had a high sensitivity and a high specificity for LS. A 92% sensitivity and a 100% specificity for LS were found when at least four of these five signs were present. CONCLUSIONS: Thirteen-megahertz ultrasound is a valuable tool for diagnosing LS. Morphological ultrasound diagnostic criteria had a high specificity and a high sensitivity. PMID- 12752131 TI - Clinicopathological and genotypic aspects of anticonvulsant-induced pseudolymphoma syndrome. AB - BACKGROUND: Pseudolymphoma syndrome (PLS) is relatively rare but can lead to death if there are extensive skin lesions, severe hepatitis, agranulocytosis and neutropenia. PLS may also give rise to harmful effects if misdiagnosed as malignant lymphoma and patients with PLS are treated unnecessarily with chemotherapy, because it may mimic histologically other lymphomas, including mycosis fungoides (MF). OBJECTIVES: To examine the clinicopathological and genotypic features of anticonvulsant-induced PLS. Patients and methods We retrospectively reviewed clinical, laboratory and histological findings for eight cases of anticonvulsant-induced PLS, and performed T-cell receptor gene rearrangement using polymerase chain reaction with paraffin-embedded specimens from each case. RESULTS: The causative agents were carbamazepine (four cases), phenytoin (two cases), phenobarbital (one case) and valproic acid (one case). A cross-reaction between phenobarbital and phenytoin was observed in one case. The duration from the start of anticonvulsant therapy to skin eruption was 3-24 weeks (mean 7 weeks). The skin lesions were generalized maculopapular eruptions in all cases, including one case accompanied by vesiculopustular lesions. The frequencies of the associated features were as follows: facial oedema (88%), fever (75%), lymphadenopathy (63%), and hepatomegaly (25%). Laboratory findings revealed leukocytosis, atypical lymphocytes, eosinophilia, monocytosis, neutrophilia, lymphocytosis and abnormal liver function. Histopathologically, there was similarity between PLS and MF in that epidermotrophism of atypical lymphocytes (100%) and Pautrier's microabscess-like structures (38%) were observed. However, PLS has some differences from MF that include moderate to marked spongiosis (75%), necrotic keratinocytes (63%), and infiltration of eosinophils (25%) in the epidermis and, in the dermis, papillary dermal oedema (100%), extravasated erythrocytes (100%), lymphocytes within the dermis larger than those within the epidermis (63%), and infiltration of various inflammatory cells including neutrophils (50%). Genotypic analysis demonstrated a rearrangement of the T-cell receptor-gamma gene in one of eight cases studied. There were no deaths and all cases were improved at 2-9 weeks (mean 6 weeks), after the cessation of causative agents, systemic and topical corticosteroid therapy, and symptomatic therapy. There were no significant differences in clinical, laboratory and histological findings between the causative agents. CONCLUSIONS: PLS may show histopathological findings similar to MF and take a prolonged course even after the cessation of causative agents. Thus, a clear understanding and diagnosis of this disease is considered to have an important effect on treatment and prognosis. PMID- 12752132 TI - Natural rubber latex allergy in a health-care population in Wales. AB - BACKGROUND: Type 1 natural rubber latex (NRL) allergy prevalence rates quoted vary due to the different populations studied and methodologies used for assessment and investigation of NRL allergy. OBJECTIVES: To investigate the rate of type 1 allergy to NRL in health-care workers (HCWs) in a single NHS trust in Wales. METHODS: A response rate of 3716 of 5548 (67%) of the employees was obtained with a latex allergy questionnaire administered by the occupational health departments. Type 1 NRL allergy was diagnosed by correlating clinical symptoms, skin prick testing and IgE RAST to latex in the dermatology department. RESULTS: The period prevalence (1998-2001) for type 1 clinical latex allergy in HCWs was 25 of 4439 (0.56%). Of the 25 positive HCWs, 18 (72%) were nurses. CONCLUSIONS: The low prevalence rate of type 1 NRL allergy should not lead to underestimating the importance of clinically significant NRL allergy and the risks that need to be minimized. PMID- 12752133 TI - Lichenoid reactions of murine mucosa associated with amalgam. AB - BACKGROUND: In 97% of all patients with oral lichenoid reactions (OLR) associated with dental amalgam a removal of the fillings leads to a decline of the lesions, as a minimum. OBJECTIVES: The aim of this study was to determine if contact allergic or local toxic effects or both may contribute to OLR using an animal model with mercury-sensitive and non-sensitive rats. METHODS: Twenty Brown Norway rats, which have a genetic predisposition for an autoimmune syndrome after exposure to mercury and 20 Lewis rats, not mercury sensitive, were treated as follows: 10 animals of each group were sensitized with a low dose of mercuric chloride. Half of all animals received local exposure of the right buccal mucosa to amalgam (left: control), the others to amalgam alloy free of mercury. All rats were patch tested with an amalgam series. RESULTS: After 20 days of exposure 96% of all animals showed white mucosal lesions restricted to the contact zone of the alloy on the treated side, but only up to 25% had a positive patch test reaction to amalgam or inorganic mercury (INM). The lesions showed no relation to species, alloy, sensitization or patch test reaction. CONCLUSIONS: While allergic mechanisms may contribute to mucosal contact lesions in Brown Norway rats, this is less probable in Lewis rats. Mercury in general appears to be irrelevant in the development of ORL in this study. If this holds true for humans as well, patch testing with an amalgam series may be helpful in a minor fraction of all patients with OLR. PMID- 12752134 TI - Histopathological diagnosis of onychomycosis by periodic acid-Schiff-stained nail clippings. AB - BACKGROUND: The current laboratory methods for diagnosing fungal infections of the nails are the potassium hydroxide (KOH) scraping technique and fungal culture. However, due to the long incubation period required for fungal culture and the reported rate of approximately 30% false negative results observed when using these methods, a quick and highly specific screening test for diagnosing onychomycosis is urgently needed. OBJECTIVES: In a prospective study, to compare the traditional mycological diagnostics using culture medium and KOH preparation with the histopathological diagnosis of onychomycosis by periodic acid-Schiff (PAS)-stained nail clippings. METHODS: Material from 387 nails of 350 patients suspected of having onychomycosis was obtained and a KOH stain as well as two fungal cultures (Kimmig agar with and without cycloheximide) were prepared. In addition, the same specimen was histopathologically examined (PAS stain). RESULTS: Culture medium and KOH preparation respectively revealed 100 and 156 cases of onychomycosis, as compared with 182 cases by histological examination. Histological examination gave a significantly higher rate of positive results (P < 0.05). Considering the total number of positive results given by at least one of the three methods (total = 438), histological evaluation was found to give the highest rate of successful recognition of mycotic infection (41.6%). CONCLUSIONS: The histopathological evaluation of PAS-stained nail clippings is very quick and easy to perform, and will increase the frequency of diagnosing onychomycotic disease above that achieved by culture and KOH preparation alone. However, because information concerning the vitality of the fungi and accurate identification of the specific pathogen is not available through this investigation alone, mycological culture continues to remain the indisputable 'gold standard' of mycological diagnostics. PMID- 12752135 TI - The expression of nectin-1alpha in normal human skin and various skin tumours. AB - BACKGROUND: A novel cell-cell adhesion system that consists of nectin and afadin has been identified at cadherin-based cell-cell adherens junctions. Nectin is a Ca2+-independent homophilic and heterophilic cell adhesion molecule that belongs to the immunoglobulin superfamily. Nectin has recently been shown to serve as an alpha-herpesvirus entry and cell-cell spread mediator. In spite of the ubiquitous expression of nectin-1alpha, its detailed localization in human skin has not been examined so far. OBJECTIVES: To investigate the localization of nectin-1alpha in normal human skin and the alteration of its expression in malignant skin tumours. METHODS: Immunohistochemistry was employed to determine the expression of nectin 1alpha and other adhesion molecules. RESULTS: We detected nectin-1alpha in normal human epidermis, follicles and eccrine ducts. Nectin-1alpha was colocalized with E-cadherin at cell-cell adherens junctions of the epidermis. The concentration of the nectin-afadin system at cell-cell adherens junctions was reduced in the early stage of malignant transformation of keratinocytes, such as in basal cell carcinomas and squamous cell carcinomas, where the cadherin-catenin system was preserved. Nectin-1alpha at cell-cell adherens junctions was reduced in human epithelial cancer cells located at the advancing border of the tumour. CONCLUSIONS: Our results showed that nectin-1alpha is located at cell-cell adherens junctions in human skin and that reduction of nectin-1alpha at cell-cell adherens junctions may be involved in the invasion of squamous cell tumours. PMID- 12752136 TI - Assessment of thickness of photoprotective lipsticks and frequency of reapplication: results from a laboratory test and a field experiment. AB - BACKGROUND: The thickness of the sunscreen layer that is actually applied by consumers under usual conditions has been determined for photoprotective lotions and creams; however, this question is still unanswered for photoprotective lipsticks. OBJECTIVES: To assess lipstick thickness (area density) and frequency of application per day for two commercially available photoprotective lipsticks with different consistency. METHODS: The study consisted of a laboratory test and a field experiment. In the laboratory test the applied lipstick thickness was determined as area density in mg cm(-2) for a group of 28 panellists under standardized conditions. In a separate group of 18 subjects we assessed the area density and the frequency of application per day for two photoprotective lipsticks during a 6-day skiing course. RESULTS: In the laboratory test the median and 95% confidence interval of the area density was 0.98 mg cm(-2) (0.66 1.65) and 0.86 mg cm(-2) (0.63-1.40) for products A and B, respectively. The respective values of the field experiment were 1.58 mg cm(-2) (0.79-2.23) (product A) and 1.76 mg cm(-2) (1.16-3.50) (product B). Only 11% of all applications of lipstick A and 6% of all applications of lipstick B reached the reference area density of 2.0 mg cm(-2). The difference between the median of the area density for lipstick A (firm consistency) and lipstick B (soft consistency) was not statistically significant. No statistically significant influence on the area density was found for age, sex, photobiological skin type or regular lipstick use. The median daily frequency of application was 2.2 times for lipstick A and 3.0 times for lipstick B. CONCLUSION: Our investigation shows that photoprotective lipsticks are applied in a much thinner layer than recommended by international standards (2 mg cm(-2)). This results in a significant reduction of the photoprotective capacity. Furthermore, the frequency of application is too low for adequate protection. Therefore, we propose that the sun protection factor (SPF) should be assessed for an area density that reflects the actual usage patterns. As long as the test protocol is not adapted to the reduced area density, photoprotective lipsticks with high and ultrahigh SPF should be recommended, especially for individuals with increased risk for the development of lip malignancies. PMID- 12752137 TI - Decreased expression of keratinocyte beta1 integrins in chronically sun-exposed skin in vivo. AB - BACKGROUND: Chronic exposure to ultraviolet (UV) radiation induces changes in the skin structure which are mostly found in the superficial dermis and at the dermal epidermal junction. Keratinocytes and fibroblasts contribute both to the synthesis and to the degradation of the molecules important for the integrity of this skin site. While several studies have reported on alterations of dermal components and of the functions of fibroblasts in vivo and in vitro after UV exposure, recent data suggested that keratinocytes could be the main skin cell type involved in the photoageing process. OBJECTIVES: In this study, we analysed the expression of two keratinocyte molecules namely, beta1 integrin (a proliferation marker) and involucrin (a differentiation marker) in sun-exposed and sun-protected facial skin of 16 healthy patients undergoing facial lifting. METHODS: Methods included histology, immunohistochemistry and quantitative reverse transcriptase-polymerase chain reaction analysis. RESULTS: Sun-exposed skin displayed the characteristic morphological and molecular features of dermal photoageing, compared with sun-protected skin, including dermal elastosis, diminished fibrillin and type VII collagen expression. Analysis of the epidermis in sun-exposed vs. sun-protected skin showed no histological differences, but dramatic changes in the expression of beta1 integrin and involucrin. In sun exposed skin, expression of beta1 integrin protein by epidermal basal cells was reduced, paralleling a downregulation of beta1 integrin mRNA, whereas involucrin protein expression was greatly enhanced in the superficial epidermal cell layers. Interestingly, the ratio between involucrin and beta1 integrin protein expression was consistently increased in sun-exposed skin sites. CONCLUSIONS: Collectively these results demonstrate that epidermal homeostasis is impaired by chronic UV exposure, and define beta1 integrin expression as a molecular marker of the epidermal photoageing process. PMID- 12752138 TI - Calcitriol vs. dithranol in combination with narrow-band ultraviolet B (311 nm) in psoriasis. AB - BACKGROUND: Narrow-band ultraviolet (UV) B (311 nm) phototherapy is an effective treatment for psoriasis. In order to reduce cumulative UV doses and to enhance clearance of psoriasis plaques, combination therapies with topical agents such as dithranol and calcipotriol have been established. OBJECTIVES: To compare the clinical efficacy, in a half-side manner, of UVB (311 nm) in combination with either calcitriol or dithranol. METHODS: Ten patients with symmetrical stable plaque psoriasis were treated with narrow-band UVB (311 nm) five times a week. In addition, topical calcitriol was applied twice daily to one arm, whereas the other arm and the rest of the body were treated once daily with dithranol. The follow-up period was at least 4 weeks. Efficacy was assessed separately for both arms prior to treatment and once weekly thereafter by a modified Psoriasis Area and Severity Index (PASI) score. The cumulative irradiation dose and the number of treatment sessions required for clearance of psoriasis lesions were determined for each patient. Additionally, all patients completed a quality of life questionnaire. RESULTS: Both treatment modalities notably reduced the PASI score. A clinical comparison of UVB (311 nm) in combination with either calcitriol or dithranol revealed no significant therapeutic differences between the regimens. CONCLUSIONS: Combination of narrow-band UVB (311 nm) therapy with calcitriol is equally effective as the combination with dithranol for the treatment of psoriasis. However, patients preferred calcitriol rather than dithranol when both quality of life and treatment acceptability were assessed. PMID- 12752139 TI - Alefacept therapy produces remission for patients with chronic plaque psoriasis. AB - BACKGROUND: Alefacept, human LFA-3/IgG1 fusion protein, is a novel biological agent currently being developed for the treatment of chronic plaque psoriasis. Alefacept selectively reduces the memory-effector T cells that have been implicated in the pathogenesis of the disease; as a result, alefacept is classified as a therapy that induces remission (so-called 'remittive' therapy). In a previously published randomized, placebo-controlled phase II study of intravenous alefacept in 229 patients with chronic plaque psoriasis, clinical improvement was observed during dosing as well as in the postdosing follow-up period. OBJECTIVES: To assess the remission period following alefacept therapy. METHODS: The time before re-treatment was required was measured in patients who were 'clear' or 'almost clear' of disease according to a physician global assessment at the end of the follow-up phase. RESULTS: In these patients, responses were sustained for a median of 10 months, and for up to 18 months. No patient reported disease rebound after cessation of alefacept. CONCLUSIONS: Alefacept is a biological agent for the treatment of chronic plaque psoriasis that provides disease-free intervals and time off drug therapy. PMID- 12752140 TI - Cutaneous polyarteritis nodosa in patients presenting with atrophie blanche. AB - BACKGROUND: The term 'atrophie blanche' is used both as a descriptive term denoting ivory-white stellate scars on the lower limbs as well as a diagnostic label synonymous with livedoid vasculitis, an ill-defined entity. Medium-sized vasculitides, such as polyarteritis nodosa (PAN), occasionally present with ulceration resulting in ivory-white stellate scarring on the lower limbs and may potentially be misdiagnosed as livedoid vasculitis. OBJECTIVES: To assess the occurrence, clinical and immunopathological features of medium-sized vasculitis in patients presenting with atrophie blanche without clinical and/or compression duplex ultrasonographic evidence of venous insufficiency. METHODS: We retrospectively evaluated patients presenting with atrophie blanche at the Department of Dermatology of Johns Hopkins Medical Institutions, from April 1996 until April 2002, following the diagnostic guidelines for leg ulcers of the Division of Immunodermatology. Deep and multiple skin biopsies were performed for histology. Investigations for underlying vasculitis, thrombophilia, nerve conduction studies and compression duplex ultrasonography of the lower extremities were performed in all patients. RESULTS: Of 29 consecutive patients presenting with atrophie blanche, six had underlying medium-sized vasculitis consistent with PAN, three of whom had previously been diagnosed to have segmental hyalinizing vasculitis/vasculopathy (livedoid vasculitis/vasculopathy) on superficial biopsies. All six patients with cutaneous PAN were women with a median age of 36.5 years (range 34-46) and with a median duration of the disease prior to diagnosis of 18 years (range 3-30). Of the six cutaneous PAN patients, four had neurological involvement evidenced by clinical symptoms and nerve conduction studies. No evidence of any other extracutaneous involvement was found. Erythrocyte sedimentation rate and tests for vasculitis and thrombophilic were normal in all six patients. None had evidence of venous insufficiency. Immunosuppressive therapy was effective in controlling PAN-associated cutaneous and neurological disease. Of the remaining 23 patients, two had antiphospholipid syndrome and one had homocystineaemia; all three also had evidence of venous insufficiency. One patient had multiple myeloma-associated type I cryoglobulinaemia and 19 patients had venous insufficiency alone. None of the non PAN patients had abnormalities in the nerve conduction studies. CONCLUSIONS: In patients presenting with atrophie blanche without evidence of venous insufficiency and thrombophilia, PAN should be excluded, particularly in the presence of mononeuritis multiplex. Repeated and deep biopsies are often necessary to reveal the accurate underlying pathology of necrotizing medium-sized vasculitis in the reticular dermis and the subcutis, especially in the setting of atrophie blanche lesions. Immunosuppressive therapy was effective in controlling the PAN-associated clinical manifestations. PMID- 12752141 TI - Expression of cyclooxygenase type 2 in lepromatous and tuberculoid leprosy lesions. AB - BACKGROUND: Leprosy is an infectious disease with two polar forms, tuberculoid leprosy (TL) and lepromatous leprosy (LL), which are dominated by T-helper (Th) 1 and Th2 cells, respectively. High concentrations of prostaglandin E2 produced by the inducible enzyme cyclooxygenase type 2 (COX-2) in LL could inhibit Th1 cytokine production, contributing to T-cell anergy. OBJECTIVES: To compare the COX-2 expression in LL and TL. METHODS: Skin biopsies from 40 leprosy patients (LL, n = 20; TL, n = 20) were used to determine by immunohistochemistry and automated morphometry the percentage of COX-2 immunostained cells. RESULTS: Most COX-2-positive cells were macrophages; their percentages in the inflammatory infiltrate located in the papillary dermis, reticular dermis and periadnexally were significantly higher in LL than TL (P < 0.001 by Student's t-test). CONCLUSIONS: The high expression of COX-2 in LL may be related to high prostaglandin production contributing to T-cell anergy. PMID- 12752142 TI - Increased procollagen alpha1(I) mRNA expression by dermal fibroblasts in melorheostosis. AB - We report a patient with melorheostosis in whom increased procollagen alpha1(I) mRNA expression and alpha1(I), alpha2(I) and alpha1(III) collagen secretion were observed in dermal fibroblasts obtained from a skin biopsy overlying the involved bone. The patient was a 53-year-old man with melorheostosis lesions over the left knee joint. Multiple pigmented macules were present on the medial aspect of the lower left leg. Hyperpigmentation of the basal keratinocytes, thick-walled vessels in the reticular dermis, and proliferation of normal-appearing collagen around the hair follicles were observed histologically. PMID- 12752143 TI - Multiple unilateral schwannomas: segmental neurofibromatosis type 2 or schwannomatosis? AB - Schwannomas are benign solitary tumours of the peripheral nerve sheaths. The occurrence of multiple schwannomas usually implies hereditary disease. The most frequent syndrome associated with multiple schwannomas is neurofibromatosis type 2 (NF2), which is defined by bilateral vestibular schwannomas. Schwannomatosis is a distinct disease characterized by multiple pathologically proven schwannomas in the absence of vestibular schwannomas. It is not currently known if the presence of multiple schwannomas confined to a limb may represent a mosaic form of NF2 or a distinct disease, because mutation analysis of these tumours is not routinely performed. We report a 31-year-old patient who presented with multiple slowly growing subcutaneous tumours on his left arm. His family history was negative for cutaneous tumours or central nervous system disease, and he did not have additional features of NF2. Magnetic resonance tomography and ophthalmological examination excluded vestibular schwannoma and eye stigmata of NF2. After resection of three tumours, histological analysis confirmed the diagnosis of benign schwannomas. Molecular genetic analysis by temperature gradient gel electrophoresis and microsatellite marker analysis demonstrated two distinct mutations of the NF2 gene (NF2) in two different schwannomas, with concomitant loss of heterozygosity in both tumours. In contrast, neither normal skin nor peripheral blood lymphocytes revealed mutations of NF2. The clinical and molecular genetic findings suggest that the diagnosis in our patient is schwannomatosis rather than segmental NF2 because the mutations found in different tumours were not identical. The possibility of a localized predisposition for the acquisition of NF2 mutations is discussed. PMID- 12752144 TI - Hodgkin Lymphoma in a patient with mycosis fungoides: molecular evidence for separate cellular origins. AB - We report the case of a man with mycosis fungoides (MF), who, 11 years after diagnosis, developed Hodgkin's disease. Although MF is associated with a higher than expected prevalence of other malignancies, including Hodgkin lymphoma, analysis of cells from the skin and lymph nodes showed findings that suggest a separate cellular origin for the two diseases. PMID- 12752145 TI - Infliximab for peristomal pyoderma gangrenosum. AB - Peristomal pyoderma gangrenosum (PPG) is a variant of pyoderma gangrenosum (PG) that is more refractory to treatment. It is a cause of severe morbidity and poses a therapeutic challenge for the clinician. Infliximab (Remicade(R); Centocor, Malvern, PA, USA) is a chimeric monoclonal antibody directed against tumour necrosis factor-alpha that has been proven to be effective in the treatment of inflammatory bowel disease (IBD) and rheumatoid arthritis. Currently, very few reports exist documenting its use in the treatment of PG and none in the treatment of PPG. We describe our experience of treating three patients with IBD associated PPG with infliximab. All patients tolerated the drug without significant side-effects. Two patients with PPG recovered completely following the administration of infliximab, and one patient had a partial response to the drug. We conclude that infliximab appears to be a safe and effective therapeutic alternative in patients with PPG. PMID- 12752146 TI - Idiopathic hypereosinophilic syndrome associated with cutaneous infarction and deep venous thrombosis. AB - We report a case of idiopathic hypereosinophilic syndrome (HES) presenting with cutaneous infarction and subsequent extensive deep vein thrombosis. The eosinophilia improved dramatically with systemic corticosteroid therapy. A variety of skin disorders have been associated with HES, although there are no previous reports of HES associated with cutaneous infarction. HES is a rare disorder characterized by a sustained overproduction of eosinophils and multisystem disease. The aetiology of the eosinophilia remains uncertain but clonal populations of abnormal T-cells producing interleukin 5 may be implicated. PMID- 12752147 TI - Periocular ecthyma gangrenosum in a diabetic patient. PMID- 12752148 TI - Activation of mast cells by silver particles in a patient with localized argyria due to implantation of acupuncture needles. PMID- 12752149 TI - Pyogenic granuloma within unilateral dermatomal superficial telangiectasia. PMID- 12752150 TI - Successful treatment of inflammatory linear verrucous epidermal naevus with topical natural vitamin D3 (calcitriol). PMID- 12752151 TI - Successful combined surgical and octreotide treatment of severe pretibial myxoedema reviewed after 9 years. PMID- 12752152 TI - The suppression of primary palmar-plantar hyperhidrosis by topiramate. PMID- 12752153 TI - The possible role of dartoic muscle degeneration in the pathogenesis of idiopathic scrotal calcinosis. PMID- 12752154 TI - Topical tacrolimus for cutaneous lupus erythematosus. PMID- 12752156 TI - Tumour necrosis factor-related apoptosis-inducing ligand expression in atopic dermatitis. PMID- 12752157 TI - Carcinoma with eccrine differentiation arising in a seborrhoeic keratosis. PMID- 12752158 TI - Topical tacrolimus (FK506) for relapsing erosive stomatitis in paraneoplastic pemphigus. PMID- 12752159 TI - Nail-destroying epithelioid haemangioendothelioma showing an erythematous scar like appearance on the finger. PMID- 12752160 TI - Phakomatosis pigmentovascularis type IIb with a patent umbilical vein and inferior vena cava hypoplasia. PMID- 12752161 TI - Pemphigus foliaceus in an 11-year-old boy with dermatomyositis: simple coincidence or familial immunological background? PMID- 12752162 TI - Water loss through the lip, nail, eyelid skin, scalp skin and axillary skin measured with a closed-chamber evaporation principle. PMID- 12752163 TI - Maternity satisfaction studies and their limitations: "What is, must still be best". AB - BACKGROUND: Health policymakers throughout the developed world are paying close attention to factors in maternity care that may influence women's satisfaction. This paper examines some of these factors in the light of observations from previous studies of satisfaction with health services. METHODS: The Scottish Birth Study, a cross-sectional questionnaire survey, sought the views of all women in Scotland delivering during a 10-day period in 1998. A total of 1,137 women completed and returned questionnaires (response rate = 69%). RESULTS: Women were overwhelmingly satisfied with their prenatal, intrapartum, and postnatal care. As is common in this type of study, reports of dissatisfaction were relatively low. However, differences occurred in satisfaction levels between subgroups; for example, the fewer the number of caregivers the woman had during childbirth, the more likely she was to be satisfied with the care received. A range of factors appeared to influence reported satisfaction levels, such as characteristics of the care provided and the woman's psychosocial circumstances. CONCLUSIONS: In addition to the inherent limitations of satisfaction studies found in the literature, problems may arise if such surveys are used uncritically to shape the future provision of maternity services, because service users tend to value the status quo over innovations of which they have no experience. Therefore, although satisfaction surveys have a role to play, we argue that they should only be used with caution, and preferably as part of an array of tools. PMID- 12752164 TI - Neonatal outcome after trial of labor compared with elective repeat cesarean section. AB - BACKGROUND: Trial of labor after cesarean section has been an important strategy for lowering the rate of cesarean delivery in the United States, but concerns regarding its safety remain. The purpose of this study was to evaluate the outcome of newborns delivered by elective repeat cesarean section compared to delivery following a trial of labor after cesarean. METHODS: All low-risk mothers with 1 or 2 previous cesareans and no prior vaginal deliveries, who delivered at our institution from December 1994 through July 1995, were identified. Neonatal outcomes were compared between 136 women who delivered by elective repeat cesarean section and 313 women who delivered after a trial of labor. To investigate reasons for differences in outcome between these groups, neonatal outcomes within the trial of labor group were then compared between those mothers who had received epidural analgesia (n = 230) and those who did not (n = 83). RESULTS: Infants delivered after a trial of labor had increased rates of sepsis evaluation (23.3% vs 12.5%, p = 0.008); antibiotic treatment (11.5% vs 4.4%, p = 0.02); intubation to evaluate for the presence of meconium below the cords (11.5% vs 1.5%, p < 0.001); and mild bruising (8.0% vs 1.5%, p = 0.008). Within the trial of labor group, infants of mothers who received epidural analgesia were more likely to have received diagnostic tests and therapeutic interventions including sepsis evaluation (29.6% vs 6.0%, p = 0.001) and antibiotic treatment (13.9% vs 4.8%, p = 0.03) than within the no-epidural analgesia group. CONCLUSIONS: Infants born to mothers after a trial of labor are twice as likely to undergo diagnostic tests and therapeutic interventions than infants born after an elective repeat cesarean section, but the increase occurred only in the subgroup of infants whose mothers received epidural analgesia for pain relief during labor. The higher rate of intervention could relate to the well-documented increase in intrapartum fever that occurs with epidural use. PMID- 12752165 TI - Do Italian mothers prefer cesarean delivery? AB - BACKGROUND: In Italy the proportion of births by cesarean section rose from 11.2 percent in 1980 to 27.9 percent in 1996 and 33.2 percent in 2000. The aim of this study was to identify factors, other than medical and obstetrical risk, that may influence the method of delivery and to analyze mother's preference for vaginal versus cesarean delivery among women after the birth of their first baby in university hospitals in Italy. METHODS: Primiparous women were selected from 100 consecutive deliveries in 23 university hospitals in 1999. To determine antenatal, delivery, and postnatal history, and women's preference for method of delivery, trained health personnel interviewed 1986 women. RESULTS: Of the 1986 women who were interviewed (response rate 95%), 1023 primiparas comprised the study sample. The cesarean section rate was 36 percent. Ninety-one percent of the women who delivered spontaneously and 73 percent of those who underwent a cesarean section would have preferred a vaginal delivery. CONCLUSIONS: Most of the interviewed women in this study preferred, or were satisfied with, vaginal birth. PMID- 12752166 TI - Do perceived attitudes of physicians and hospital staff affect breastfeeding decisions? AB - BACKGROUND: In the United States, since a substantial percentage of mothers are not breastfeeding, research is needed to assess important influences on breastfeeding. The current study assessed the impact on breastfeeding of the perceived attitudes of health care providers about infant feeding. METHODS: A longitudinal mail survey (1993-1994) was administered to 1620 women prenatally through 12 months postpartum; the current study focused on the prenatal and neonatal periods (66% response rate). The outcome variable was failure to breastfeed beyond 6 weeks. Predictor variables were the mother's perceptions of her prenatal physician's and hospital staff's attitudes on infant feeding. Analysis controlled for mother's prenatal breastfeeding intentions, father's feeding preference, and demographic and psychosocial variables. RESULTS: Forty one percent of the mothers were not breastfeeding at 6 weeks postpartum. Substantial percentages of mothers reported that physicians and hospital staff expressed a preference for breastfeeding (38% and 57%, respectively), or expressed no preference (61% and 42%, respectively), whereas few favored formula feeding. Adjusted analyses indicated that "no preference" by hospital staff was a significant risk factor for failure to breastfeed beyond 6 weeks. "No preference" by physicians did not significantly influence breastfeeding outcome in these analyses. Further analyses indicated that the effects of perceived hospital staff attitudes were only present for mothers who intended prenatally to breastfeed for 2 months or less. CONCLUSIONS: Many women did not report receiving positive breastfeeding messages from their health caregivers and hospital staff. A perceived neutral attitude from the hospital staff is related to not breastfeeding beyond 6 weeks, especially among mothers who prenatally intended to breastfeed for only a short time. PMID- 12752167 TI - A simplified predictive index for the detection of women at risk for postnatal depression. AB - BACKGROUND: Many women who suffer from postnatal depression are never diagnosed or treated. The objective of this study was to develop an index for use in maternity settings that identifies women who may be at risk for postnatal depression. METHODS: Women (n = 1762) attending the "booking-in" clinic were screened for antenatal risk factors for postnatal depression. On the third postnatal day eligible women were screened for postnatal risk factors. The Edinburgh Postnatal Depression Scale was mailed to participants 16 weeks after the birth. A predictive index was developed, based on the mean Edinburgh Postnatal Depression Scale scores for each risk factor. The sensitivity, specificity, positive predictive value, and negative predictive value were used to assess the diagnostic value of the index. RESULTS: Seven hundred and twenty three (50.1%) of the eligible women completed all phases of the study. Of this group, 93 (12.2%) women scored higher than 12 on the Edinburgh Postnatal Depression Scale. At a cutoff of 6, the index had positive predictive value of 39.8 percent for postnatal depression, a threefold improvement over the base rate. CONCLUSION: The Brisbane Postnatal Depression Index provides a clinically useful method for identifying women at risk for developing postnatal depression. It has applications for early intervention or to identify high-risk groups for research purposes. PMID- 12752168 TI - Women's satisfaction with their involvement in health care decisions during a high-risk pregnancy. AB - BACKGROUND: Increasingly, women seek involvement in decisions about their health care. The purpose of this study was to examine women's experience of, and satisfaction with, their involvement in health care decisions during a high-risk pregnancy. METHODS: Forty-seven women with hypertension or threatened preterm delivery (including multiple births) were interviewed after the birth of their child. They received prenatal care at home from nurses in a community program or were hospitalized. The in-depth interviews were audiotaped and transcribed; data were analyzed using constant comparative methods. RESULTS: Women identified an increased feeling of responsibility for the health of their baby and themselves, but differed in choosing active or passive involvement in health care decisions. Women who wanted active involvement achieved it through one of three processes: struggling for, negotiating, or being encouraged. Women who wanted passive involvement and women facing health crises used the process of trusting in the expertise of nurses and physicians. Women were satisfied if the care from health care professionals was congruent with how they wanted to be involved in decision making. CONCLUSIONS: Although most women want to be actively involved in health decision-making during a high-risk pregnancy, some prefer a passive role. The setting of prenatal care, community-based or in-hospital, was less important than the ability of nurses and physicians to support the woman in her preferred role in decision-making. PMID- 12752169 TI - Taking antenatal group B Streptococcus seriously: women's experiences of screening and perceptions of risk. AB - BACKGROUND: Early-onset group B streptococcal disease is a serious cause of neonatal morbidity and mortality. Although screening protocols for group B streptococcus are common, little is known of women's perceptions of this screening and the disease itself. The purpose of this study was to gain an understanding of women's experiences, knowledge, and perceptions about this bacteria and its screening. METHODS: Nine focus group interviews with 35 women explored their experiences and understanding of group B streptococcus screening. Transcribed interview data were interpreted to identify and articulate the women's experiences. RESULTS: Most women had little knowledge or understanding of group B streptococcus, obtaining their information largely from the stories or experiences of friends or family. Women struggled to understand the meaning and implications, both physical and "moral," of the disease for their baby and for themselves, clearly indicating both the subjective and statistical importance of the concept of risk for pregnant women. CONCLUSIONS: Group B streptococcus continues to be poorly understood by pregnant women who try to understand and weigh up its risks and implications so as to make the best decisions about screening. The women participated in screening ultimately, however, since it was seen to be patently "best for baby," relatively easy for them to undergo, and part of routine antenatal care. PMID- 12752170 TI - "Spin doctoring" the research. PMID- 12752171 TI - Maternal positions and pelves revisited. PMID- 12752175 TI - To question or not to question? PMID- 12752176 TI - Brachial plexus palsy causation. PMID- 12752179 TI - Ticks in australia. AB - Ticks are blood-sucking parasites of vertebrates that may embed in human skin and are therefore of clinical relevance to dermatologists and their medical colleagues. Depending on the species involved, consequences of tick attachment vary from minor local reactions to significant systemic sequelae. It is possible to minimize morbidity by removing the tick in its entirety as soon as it is detected. Some techniques to achieve this are described. This review will aid clinicians in the recognition and practical management of tick bites in Australia. PMID- 12752180 TI - Diagnosis and treatment of the common idiopathic photodermatoses. AB - The idiopathic photodermatoses are the most common cause of photosensitivity and the commonest of these are polymorphic light eruption, actinic prurigo, chronic actinic dermatitis and solar urticaria. The clinical presentation, investigation and treatment of these four disorders are presented. Spontaneous improvement does occur. PMID- 12752181 TI - Treatment of acne with isotretinoin: recommendations based on Australian experience. AB - Oral isotretinoin revolutionized acne treatment when it was introduced in 1982 in the USA. However, its use was restricted to patients with severe nodulocystic acne. Today its use worldwide has expanded to treat also patients with less severe but scarring acne who are responding unsatisfactorily to conventional therapies. These recommendations assess the potential for use of oral isotretinoin as a safe and effective treatment for severe nodulocystic acne unresponsive to conventional therapy, and acne of any severity that causes scarring or is associated with psychological distress. PMID- 12752182 TI - PUVA treatment of alopecia areata totalis and universalis: a retrospective study. AB - The results of PUVA treatment of alopecia areata (AA) totalis and universalis were reviewed in 26 adult patients. Eight of 15 patients with AA totalis and six of 11 patients with AA universalis achieved a complete response (>90% hair regrowth). Patients with AA totalis had a greater incidence of treatment failure (<25% hair regrowth) than those with AA universalis. Patients with a family history of AA were significantly less likely to have a positive response to PUVA than those with no family history. Sex, age at diagnosis and treatment, interval between diagnosis and treatment, and background of atopy were not significant determinants of outcome. Although unable to show significance for clinical response to treatment, this study demonstrates complete hair regrowth in patients with both AA totalis (53%) and universalis (55%) while reporting a low relapse rate among these patients (21%) within a long period of follow up (mean 5.2 years). PMID- 12752183 TI - Clinical study of primary cutaneous B-cell lymphoma using both the European Organization for Research and Treatment of Cancer and World Health Organization classifications. AB - Primary cutaneous B-cell lymphoma (PCBCL) is rare, with few series reported in the literature. Its classification and treatment remain controversial. Biopsy specimens of 13 patients with PCBCL were classified according to both the European Organization for Research and Treatment of Cancer (EORTC) and the new World Health Organization (WHO) classifications. Treatment and clinical outcomes were documented. Using the EORTC classification there were seven men and six women aged 32-85 years (mean = 51 years) with follicle centre cell (FCC) lymphoma (nine), immunocytoma (two) and primary cutaneous large B-cell lymphoma of the leg (PCLBCL-leg) (two). When the WHO classification was used, the nine patients with FCC were reclassified as follicle centre (five) and diffuse large B-cell lymphoma (four). Most patients had localized disease (12). Initial treatment consisted of radiotherapy alone (seven), combination chemotherapy alone (one), combined chemoradiotherapy (three) and surgery (two). Twelve patients achieved complete remission (median follow up 28 months, range 10-167 months). One patient with PCLBCL-leg died from progressive cutaneous disease. Most localized PCBCL lesions (except PCLBCL-leg) have a favourable prognosis. We recommend that clinicians be familiar with the important differences in the EORTC and WHO classifications. Further large prospective studies comparing the WHO and EORTC classifications are required to more clearly delineate the outcomes of the increasing number of patients who are classified as DLBCL by the WHO system. PMID- 12752184 TI - Treatment of severe recalcitrant plaque psoriasis with single-dose intravenous tumour necrosis factor-alpha antibody (infliximab). AB - Infliximab is a chimeric anti-tumour necrosis factor-alpha antibody that has been demonstrated to have marked efficacy in the treatment of psoriasis. Seven patients with chronic plaque psoriasis were treated with single-dose intravenous infliximab (5 mg/kg), and the Psoriasis Area and Severity Index (PASI) and Dermatology Life Questionnaire Index (DLQI) were used as a measure of treatment efficacy. There was an average improvement in PASI scores of 69% at 2 weeks post infusion. There was an improvement in DLQI of 61%. Four of the seven patients were also seen at 10 weeks post infusion and the improvement in PASI and DLQI was sustained. All patients tolerated the initial infusion well without adverse events. The results indicate that single-dose infliximab is an effective and efficacious therapy for recalcitrant psoriasis and has a prolonged therapeutic effect. PMID- 12752185 TI - Genetic testing of the family with a Carney-complex member leads to successful early removal of an asymptomatic atrial myxoma in the mother of the patient. AB - Carney complex is a rare cardiocutaneous syndrome with an autosomal-dominant inheritance pattern. Apart from its cutaneous manifestations of multiple blue naevi and lentigines, it can involve multiple other organ systems, particularly the heart, where myxoma tumours commonly develop and can potentially lead to serious complications such as cerebrovascular accidents and myocardial infarction. Recently, a specific mutation in the gene encoding the R1-alpha regulatory subunit of cyclic adenosine monophosphate-dependent protein kinase A (PRKAR1alpha) has been discovered and found to be associated with a high risk of developing cardiac myxomas. We report the case of a Carney-complex family member who displayed no observable clinical or cardiac features of the disease but who was found to be positive for the PRKAR1alpha gene mutation on genetic testing. Further evaluation of this patient subsequently led to the discovery of a 3-cm atrial myxoma that had previously been undetected on cardiac assessment. This case highlights the potential benefits of using genetic screening for this disease. PMID- 12752186 TI - Treatment of Bowen's disease and basal cell carcinoma of the nose with imiquimod 5% cream. AB - We report the successful use of topical imiquimod 5% cream for extensive multifocal, recurrent (post cryotherapy), biopsy-proven Bowen's disease of the nose. Treatment was applied on a once-a-day regimen, and a total of 32 applications over 9 weeks were used. A florid local skin reaction occurred early in the treatment, necessitating a rest period and decreasing the frequency of application. The Bowen's disease was coexistent with a multifocal superficial basal cell carcinoma (BCC) that had a partial response. Persistent BCC at 4 weeks post treatment was surgically excised. This tumour showed an unusual histological picture, with normal epidermis overlying residual BCC in the papillary dermis. The Bowen's disease remains clinically clear at 12-months follow up. PMID- 12752187 TI - Recurrent morphoeic basal cell carcinoma at the lateral canthus with orbitocranial invasion. AB - We present a case of recurrent, nodulocystic and morphoeic basal cell carcinoma (BCC) at the lateral canthus, with clinically silent orbital and intracranial invasion which was treated with radiotherapy. Orbital invasion is a rare complication of periocular BCC and occurs primarily in high-risk canthal lesions. Preoperative ophthalmological review and imaging should be considered in these cases. PMID- 12752188 TI - Cutaneous Wegener's granulomatosis: a variant or atypical localized form? AB - A 74-year-old woman presented with an antineutrophil cytoplasmic antibody titre negative, treatment-responsive Wegener's granulomatosis confined to the integument. She initially presented with a painful left postauricular nodulo ulcerative lesion with chronically discharging sinuses. This lesion was effectively treated with a short, 3-month course of cyclophosphamide and 24 months of oral prednisone. After 5 months in remission, she developed further similar ulcers, in addition to painless nodules on her ankles and feet bilaterally. These lesions resolved with an extra 32 months of high-dose oral prednisone therapy before complete remission. At most recent review, there was no evidence of disease recurrence 21 months after ceasing all active treatment. Histology demonstrated a granulomatous inflammation. No systemic disease progression to the upper respiratory tract, lung or kidney was detected. This case highlights the importance of being aware of atypical or partial presentations of Wegener's granulomatosis. This diagnosis needs to be considered with patients presenting with a culture-negative chronic ulcer, where malignancy and trauma have been excluded. This will avoid unnecessary surgery and ensure early diagnosis and effective treatment of a disease that is disfiguring and usually fatal if inappropriately treated. PMID- 12752189 TI - Multiple squamous cell carcinomas complicating linear porokeratosis. AB - Linear porokeratosis is a distinctive variant of porokeratosis with malignant transformation, a known but infrequently reported complication. We report a case of linear porokeratosis on the right leg of a 57-year-old man present since infancy that, over a 2-year period, developed two squamous cell carcinomas within the lesion. Concurrent right inguinal lymphadenopathy was demonstrated on biopsy to be reactive in nature. PMID- 12752190 TI - Pachydermodactyly: a forme fruste of knuckle pads? AB - A 16-year-old boy presented with painless swellings localized to the radial and ulnar aspects of his second through to the fifth fingers on the left hand, with more subtle changes affecting two fingers on the opposite hand. This had developed in the absence of mechanical trauma. Investigations for an arthropathy were negative, while a biopsy showed marked epidermal hyperplasia and an expanded dermis. These features are typical of pachydermodactyly, a benign dermatosis of uncertain aetiology. The interesting feature of this case is the presence of knuckle pads in the father of the patient, which raises the possibility that these two similar entities are related. PMID- 12752191 TI - Pedigree of multiple benign adnexal tumours of Brooke-Spiegler type. AB - A pedigree of autosomal dominant expression of multiple benign adnexal tumours is presented. Seven cases spanning three generations are discussed. The clinical manifestations of these tumours are quite variable, including multiple papules concentrated on the face, scalp nodules and a large turban tumour. One member of the family had a linear papular eruption involving one half of his body. Histopathology of all lesions demonstrated benign adnexal characteristics, including well-characterized eccrine spiradenomas, trichoepitheliomas and an eccrine cylindroma. The cutaneous tumours occurring in these patients have continued to develop during their lifetimes. The authors propose that this pedigree has phenotypic characteristics consistent with Brooke-Spiegler syndrome. PMID- 12752192 TI - Pigmented fungiform papillae on the tongue in an Asian man. AB - Fungiform papillae of the tongue vary in their appearance. It is not rare for them to be pigmented in black subjects, but this is less common in Asians. We report a case of pigmented fungiform papillae in a 65-year-old Vietnamese man. The patient also had recurrent aphthous ulceration and hepatitis C. Pigmentation affected all fungiform papillae and was asymptomatic. Macrocytosis was noted on full blood examination, while liver function tests were normal. The pigmentation of the fungiform papillae remained stable. PMID- 12752194 TI - Contact dermatitis to Shiitake mushroom. AB - A 44-year-old woman with occupational allergic contact dermatitis to Shiitake mushroom (Lentinus edodes), sawdust and thiuram. She presented with an 8-month history of hand and face dermatitis that developed after working for 2.5 years in a mushroom farm. Within 3 months of ceasing work at the farm, her symptoms resolved completely. PMID- 12752193 TI - Reactive angioendotheliomatosis in the setting of antiphospholipid syndrome. AB - A 31-year-old man with systemic lupus erythematosus and antiphospholipid syndrome developed erythematous purpuric plaques distributed over the lower chest, abdomen and upper thighs. Biopsy of lesional skin revealed intravascular proliferation of endothelial cells with associated microthrombi formation. The histological pattern was consistent with reactive angioendotheliomatosis, a rare reactive pattern seen associated with disparate medical conditions. The pathogenesis of the reactive angioendotheliomatosis in our patient was suspected to be related to his procoagulant state; thrombi formed despite a therapeutic international normalized ratio while on warfarin. His lesions began to resolve with the cessation of warfarin and commencement of subcutaneous enoxaparin, oral clopidogrel and oral aspirin. The skin biopsy findings were pivotal in influencing the change of therapy in this patient and decreasing his immunosuppression. PMID- 12752196 TI - Proceedings of the 2nd Latin American Congress for Artificial Organs and Biomaterials. Belo Horizonte, Brazil, December 5-8, 2001. PMID- 12752197 TI - New concepts in peritoneal dialysis: new wine in old barrels? PMID- 12752198 TI - Effect of process parameters on the characteristics of porous calcium phosphate ceramics for bone tissue scaffolds. AB - Porous hydroxyapatite (HA) has already been widely used as a bone substitute due to its similarity with the mineral part of the bone. In this work, cylindrical tablets with micro and macro porosity were produced from stoichiometric and deficient hydroxyapatites by using naphthalene as porosifier agent. The influence of the processing parameters such as Ca/P ratio of start material, calcination temperature, and naphthalene content on the characteristics of porous calcium phosphate tablets was evaluated. Three mineral phases-HA, alpha-TCP (alpha tri calcium phosphate), and beta-TCP (beta tricalcium phos-phate)-with variable contents were identified by x-ray diffraction (XRD) and Fourier-transformed infrared spectroscopy (FT-IR). Image analysis and density measurements were used to characterize sample porosity. As expected, the total porosity of the calcinated material is not dependent on the stoichiometry of the precursor hydroxyapatite. For calcium-deficient hydroxyapatite, the increase in naphthalene content contributes to stabilize alpha-TCP phase, altering the relative phases content. PMID- 12752199 TI - Dual-setting calcium phosphate cement modified with ammonium polyacrylate. AB - alpha-Tricalcium phosphate bone cement, as formerly designed and developed by Driessens et al., consists of a powder composed by alpha-tricalcium phosphate (alpha-TCP) and hydroxyapatite (HA) seeds, and an aqueous solution of Na2HPO4 as mixing liquid. After mixing powder and liquid, alpha-TCP dissolves into the liquid and calcium deficient hydroxyapatite (CDHA), more insoluble than the former, precipitates as an entanglement of crystals, which causes the setting and hardening of the cement. alpha-TCP bone cement offers several advantages in comparison to calcium phosphate bioceramics and acrylic bone cements as bone graft and repairing material, like perfect adaptability to the defect size and shape, osteotransductibility, and absence of thermal effect during setting. The main handicap is its low mechanical strength. Therefore, approaching its mechanical strength to that of human bone could considerably extend its applications. In the present work, an in situ polymerization system based on acrylamide (AA) and ammonium polyacrylate (PA) as liquid reducer was added to alpha-TCP cement to increase its mechanical strength. The results showed that the addition of 20 wt% of acrylamide and 1 wt% AP to the liquid increased the compressive and tensile strength of alpha-TCP bone cement by 149 and 69% (55 and 21 MPa), respectively. The improvement in mechanical strength seems to be caused by a decrease of porosity and the reinforcing effect of a polyacrylamide network coexisting with the entanglement of CDHA crystals. The studied additives do not affect the nature of the final product of the setting reaction, CDHA, but promote the reduction of its crystal size. PMID- 12752200 TI - The monomeric formulation optimization of dental composite: mechanical and kinetic studies. AB - The optimization of the monomer composition, using a polymerization kinetic study and the mechanical behavior study of the composites, was the principal objective of this work. The monomeric mixture analyzed is a part of the composite that has been in development at the University of Campinas, Brazil. Parameters such as initiator, accelerator, and macro-components composition were optimized. The infrared spectroscopy was the analytical technique used in the kinetic study. On the mechanical characterization, the stress strain, flexure stress and its modulus, were the properties analyzed. The dependency between the polymeric reaction and the physical behavior of the composites and the monomer composition was observed and studied. Finally, the monomer optimum composition was determined. PMID- 12752201 TI - Biocompatibility study of polymeric biomaterials. AB - Polymeric hydrogels are used as wound dressing material since these materials show advantages such as pain relief, exudates absorption, barrier to microorganisms, permeability, and others. This article shows the results obtained in a study aiming to know the biological performance of different polymeric materials to be used in contact with skin: PVP hydrogels and acrylate adhesive. The biocompatibility was determined by in vitro assay of cytotoxicity and in vivo assay by using the contact test of irritability in rabbits. All the tested samples presented no toxicity and no dermal irritation. PMID- 12752202 TI - Bioreabsorbable polymer scaffold as temporary meniscal prosthesis. AB - Menisci have an important role in load bearing, shock absorption, knee joint stability, and joint lubrication. Meniscal lesions and meniscectomy are followed by osteoarthritis in a high percentage of patients. At present, there is no ideal prosthesis for meniscal substitution. In this work, a bioreabsorbable polymer scaffold made of poly(L-lactic acid) (PLLA) and poly(p-dioxanone) (PPD) blend was developed to be used as a temporary meniscal prosthesis to stimulate the formation of an in situ meniscal replication while the scaffold is reabsorbed by the organism. Total meniscectomy of medial meniscus and arthrotomy was made in both back knees of 34 adult New Zealand white rabbits by medial parapatellar incision. The scaffolds were sutured in one of the knees, and other was used as a control. A meniscal replica was developed, suggesting that this material has great potential to be used as a meniscal prosthesis, especially because the new meniscus promoted a significant protection of cartilage, and cartilage degeneration in the control condyles was observed. PMID- 12752203 TI - Structure and dosimetric analysis of biodegradable glasses for prostate cancer treatment. AB - This work analyzes SiO2 and SiO2-CaO glasses incorporated with samarium atoms produced by sol-gel synthesis. The goal is to provide biocompatible and biodegradable radioactive seeds as an alternative to be used in brachytherapy for the treatment of prostate cancer. The chemical and physical characteristics of the obtained glasses were analyzed by energy dispersive x-ray spectroscopy, x-ray diffraction, He picnometry, and nitrogen adsorption analysis. A theoretical analysis of the process of neutron activation of the samples was also conducted through the calculation of the activity of the seeds and the beta- and gamma-ray doses emitted by the seeds. The results demonstrate the incorporation of samarium atoms in the glass matrix. The experimental data coupled with the theoretical studies in neutron activation suggest that it is possible to obtain radioactive seeds with activities equivalent to 125I seeds used in prostatic brachytherapy. PMID- 12752204 TI - Biomimetic mineralization of charged collagen matrices: in vitro and in vivo study. AB - Polyanionic collagen matrix prepared by hydrolysis side chain amides of asparagine and glutamine was mineralized in vivo, without inflammatory response, biodegradation, or resorption, with calcium phosphate deposited in close resemblance to the D-periodicity of collagen fibrils assembly. In vitro results with the same material produced mineralized collagen fibers with a similar morphology and chemical characteristics, suggesting that amide hydrolysis may have introduced into this matrix, signs for the controlled mineralization of collagen fiber. TEM indicated that amide hydrolysis occurred near the OVERLAP and GAP zones, as suggested by the significant reduction in inter-band distances in these regions. The lack of an inflammatory response associated to the similar mineralization pattern observed in vivo and in vitro suggests not only the biomimetic behavior of polyanionic collagen matrix, but also its potential uses as scaffold for bone tissue reconstruction. Based on these results, a model for the in vitro mineralization was also proposed. PMID- 12752205 TI - Digital image processing for biocompatibility studies of clinical implant materials. AB - The use of computer vision coupled with scanning electron microscopy (SEM) was used to monitor the platelet adhesion and activation onto blood-contacting materials. The interaction of blood platelets with polyethylene (PE), poly(ethylene terephthalate) (PET), and poly(vinylchloride) (PVC) after contact of the polymeric surfaces with whole blood was studied. The SEM images (SEM Phillips XL 30) were captured using HLImage++ computer vision systems. A library with a considerable number of acceptance or rejection of samples has been conceived and implemented. The obtained results make the developed computational vision system a promising tool for the evaluation of blood compatibility of biomaterials. PMID- 12752206 TI - Atomic force microscopy as a tool for biomedical and biotechnological studies. AB - This work presents different applications in progress with the aid of the atomic force microscopy (AFM) technique for biomedical and biotechnological applications, comprising both the acquisition of three-dimensional images and spectroscopic force measurements, in the following systems: first, low-density lipoprotein (LDL)-glycosaminoglycans; second, lectins-polysaccharides; third, mycobacterium leprae cellular wall and Vesicular Stomatites Virus (VSV) with fibronectin laminin, and lipidic membranes; fourth, DNA-complex; and fifth, actin, as well as the development of surface functionalizing protocols and image restoration by means of mathematical techniques. PMID- 12752207 TI - Improvement on the auxiliary total artificial heart (ATAH) left chamber design. AB - The auxiliary total artificial heart (ATAH) is an electromechanically driven artificial heart with reduced dimensions, which is able to be implanted in the right thoracic or abdominal cavities of an average human patient without removing the natural heart or the heart neurohumoral inherent control mechanism for the arterial pressure. This device uses a brushless direct current motor and a mechanical actuator (roller screw) to move two diaphragms. The ATAH's beating frequency is regulated through the change of the left preload, based on Frank Starling's law, assisting the native heart in obtaining adequate blood flow. The ATAH left and right stroke volumes are 38 ml and 34 ml, respectively, giving approximately 5 L/min of cardiac output at 160 bpm. Flow visualization studies were performed in critical areas on the ATAH left chamber. A closed circuit loop was used with water and glycerin (37%) at 25 degrees C. Amberlite particles (80 mesh) were illuminated by a 1 mm planar helium-neon laser light. With left mean preload fixed at 10 mm Hg and the afterload at 100 mm Hg, the heart rate varied from 60 to 200 bpm. Two porcine valves were used on the inlet and outlet ports. The flow pattern images were obtained using a color micro-camera and a video recorder. Subsequently, these images were digitized using a PC computer. A persistent stagnant flow was detected in the left chamber inlet port. After improvement on the left chamber design, this stagnant flow disappeared. PMID- 12752208 TI - Endurance test on a textured diaphragm for the auxiliary total artificial heart (ATAH). AB - We performed an endurance test on a textured diaphragm made of polyurethane (BioSpan, The Polymer Technology Group, San Francisco, CA, U.S.A.) to be used in the auxiliary total artificial heart (ATAH), an electromechanical device that can be totally implantable without removing the natural heart due to the device's reduced dimension. The objective of this endurance test was to predict whether this diaphragm would be capable of resisting in vivo tests with the ATAH implanted for fifteen days in calves. In this study, a mock loop system simulating the human circulatory system was used. The test protocol was elaborated to reproduce extreme physiological conditions. The technique to produce the textured diaphragms made of polyurethane is shown. The textured surface is used as basis to fix a layer of calf-skin gelatin. The technique used to make the diaphragm guaranteed a totally textured surface without cracking. The diaphragm demonstrated enough resistance to be used at the 15 day in vivo experiments. PMID- 12752209 TI - Metallic biomaterials TiN-coated: corrosion analysis and biocompatibility. AB - Corrosion processes due to contact with the physiological environment should be avoided or minimized in orthopedic implants. Four metallic substrates frequently used as biomaterials: pure Ti, Ti-6Al-4V alloy, ASTM F138 stainless steel, and Co Cr-Mo alloy, were coated with TiN using the physical vapor deposition (PVD) technique. These coatings have been screened by polarization curves in physiological solutions. TiN prepared by PVD is efficient as coating for stainless steel. On titanium and alloy there are no benefits concerning the corrosion resistance compared to the bare Ti-materials. TiN coatings have been screened according to ISO 10993 standard tests for biocompatibility and exhibited no cytotoxicity, dermal irritation, or acute systemic toxicity response. PMID- 12752210 TI - Heat exchangers for cardioplegia systems: in vitro study of four different concepts. AB - The aim of this work is the evaluation of four different heat exchangers used for myocardium during cardioplegic system in cardiac surgeries. Four types of shell and tube heat exchangers made of different exchange elements were constructed, as follows: stainless steel tubes, aluminium tubes, polypropylene hollow fiber, and bellows type. The evaluation was performed by in vitro tests of parameters such as heat transfer, pressure drop, and hemolysis tendency. The result has shown that all four systems tested were able to achieve the heat performance, and to offer low resistance to flow, and safety, as well as have low tendency to hemolysis. However, we can emphasize that the bellows type heat exchanger has a significant difference with regard to the other three types. PMID- 12752211 TI - Nano-assembled films for taste sensor application. AB - An artificial taste sensor based on different types of ultra-thin films of conducting polymers (a special class of plastics that can conduct electricity) and their mixture with a lipid-like material has been able to mimic the human palate. In addition, this "electronic tongue" has been successfully employed in the analysis of tastants, suppression effects, and commercial beverages throughout AC measurements (impedance spectroscopy) in a relatively low-cost, simple, and efficient way. PMID- 12752212 TI - The application of shape memory actuators in anthropomorphic upper limb prostheses. AB - In recent years, single crystal Cu-Al-Ni alloys with shape memory behavior (SMB) became generally commercialized. They achieved the level of extended application, including upper limb human prosthesis with anthropomorphic characteristics. An actuator based in single crystal Cu-Al-Ni alloy was tested as a prototype for prosthetic actuators. Their thermal cycle times remarkably define the actuator dynamics and the idea of preheating to reduce its response time was tested. To elaborate the heating conditions, the chemical composition of martensitic and austenitic single crystals, Cu-Al-Ni alloy samples were examined. The dynamic response of a martensitic actuator made with SMB and the power consumed with preheating was analyzed. It demonstrates that the presence of more elements in alloys may be fundamental to displace the heating diagram and to reduce the power consumed. PMID- 12752213 TI - Modeling and simulation of pulsatile blood flow with a physiologic wave pattern. AB - This article presents a computational fluid dynamics (CFD) model to analyze pulsatile blood flow in a human artery. The model assumes the vessel as being a straight wall tube, and the blood flow is considered to be incompressible, Newtonian, and axisymmetric. Physiologically realistic resting conditions were adopted for the simulation tests. The main objective was to identify regions where pulsatile effects were significant. Velocity profiles were obtained for both normal and stenosed vessels. The results have shown that, for normal vessels, pulsatile effects prevail close to the vessel wall. The presence of stenosed sections resulted in flow alterations, with flow recirculation areas changing both size and location during the cardiac cycle. These results are especially important in the design of artificial organs. The knowledge of the flow pattern in complex geometries such as artificial hearts, prosthetic valves, and ventricular assist devices can help avoid or minimize thrombogenesis and hemolysis. PMID- 12752214 TI - In vitro effects of a chlorhexidine controlled delivery system. AB - The aim of this work was study the effect of the chlorhexidine : hydroxypropyl beta-cyclodextrin (CLX : HP-beta-CD) inclusion compound (IC) on in vitro slabs of bovine dentine. The substantivity, antimicrobial activity, and morphological effect of this inclusion compound were evaluated. Cyclodextrin improves the physical-chemical and pharmacological properties of drugs. Fragments of bovine dentine were immersed into either IC serial solutions at 0.24%, 0.12%, 0.06%, 0.03%, 0.015%, and 0.008% or controls water and free chlorhexidine. The desorption kinetics showed that CLX : HP-beta-CD compound release CLX for 6 days in a rate flow near to zero-order profile in comparison to plain CLX. Antimicrobial activity tests showed that CLX : HP-beta-CD inhibited A. actinomycetemcomitans and S. mutans significantly. The morphological effect studied by scanning electron microscopy (SEM) showed that CLX : HP-beta-CD did not cause morphological changes to the slab's surface. It is concluded that the chlorhexidine : hydroxypropyl-beta-CD inclusion compound creates an effective controlled release system with biological activity and that it may act as a good prevention and control agent of caries and periodontal disease in vivo. PMID- 12752215 TI - The effect of cyclodextrins on the in vitro and in vivo properties of insulin loaded poly (D,L-lactic-co-glycolic acid) microspheres. AB - In this work we describe the development and characterization of a new formulation of insulin (INS). Insulin was complexed with cyclodextrins (CD) in order to improve its solubility and stability being available as a dry powder, after encapsulation into poly (D,L-lactic-co-glycolic acid) (PLGA) microspheres. The complex INS : CD was encapsulated into microspheres in order to obtain particles with an average diameter between 2 and 6 microm. This system was able to induce significant reduction of the plasma glucose level in two rodent models, normal mice and diabetic rats, after intratracheal administration. PMID- 12752216 TI - Modulation of the pathology of late xenograft rejection by PAF-antagonist UR 12670 in the hamster-to-rat liver xenotransplant model. AB - PAF antagonists have been used in xenotransplantation to alleviate the pathogenesis of hyperacute rejection. This study evaluated the ability of the PAF antagonist UR-12670 to improve graft function in late xenograft rejection (LXR) in an orthotopic liver xenotransplantation model, and the involvement of PAF (platelet activating factor) in this type of rejection. The recipients of a hamster xenograft received standard immunosuppression (tacrolimus 0.2 mg/kg/30 days, MMF 25 mg/kg/8 days). Study groups: group A, without UR-12670, group B, UR 12670 (20 mg/kg/8 d) and group C, continuous administration of UR-12670 (20 mg/kg/d). Serum levels of xenoantibodies were evaluated by flow cytometry and tissue deposits by immunofluorescence. Immunoblot and indirect immunofluorescence assessed specificity of xenoantibodies. Conventional histology was performed. Continuous administration of UR-12670 improved the histological pattern of liver xenografts, especially necrosis, loss of hepatocytes, hemorrhage, sinusoidal congestion and lymphocyte infiltration. There was not a shift in specificity of xenoantibodies at different times posttransplantation, as demonstrated by immunoblotting and indirect immunofluorescence. UR-12670 administration had a beneficial effect on graft function and considerably improved the histopathological pattern, but it failed to induce tolerance after withdrawal of immunosuppression. UR-12670 had an immunomodulatory effect on cellular response but not on antibody production. There was not a change in the specificity of xenoantibodies produced at LXR compared with pretransplant antibodies. PMID- 12752217 TI - Immunohistochemical analysis of Smac/DIABLO expression in human carcinomas and sarcomas. AB - Second mitochondria-derived activator of caspases (Smac/DIABLO) is released from mitochondria into the cytosol during apoptosis, promoting caspase activation by neutralizing the inhibition of inhibitor of apoptosis proteins (IAPs) on caspases. Alteration of apoptosis is essential for cancer development, and cancer cell death by radiation and chemotherapy is largely dependent upon apoptosis. In this study, archival tissues of 100 carcinomas and 50 sarcomas from various origins were analyzed by immunohistochemistry for the expression of Smac/DIABLO. Smac/DIABLO immunoreactivity was seen in 62 of 100 (62%) carcinomas, including 42 of 60 stomach carcinomas, 7 of 10 colorectal carcinomas, 4 of 10 lung carcinomas, 7 of 10 ovarian carcinomas, and 2 of 10 prostate carcinomas. Smac/DIABLO is expressed in 11 of 50 (22%) sarcomas, including 2 of 8 malignant schwannomas, 5 of 11 rhabdomyosarcomas, 2 of 7 malignant fibrous histiocytomas, 1 of 6 leiomyosarcomas, 0 of 8 angiosarcomas, 0 of 8 liposarcomas, and 1 of 2 Ewing's sarcomas. These data demonstrated that Smac/DIABLO expression levels vary depending on the individual cancer types. Furthermore, the present study showed that many human cancers do not express Smac/DIABLO, and suggest that lack of Smac/DIABLO expression in the cancer cells may inhibit apoptosis, thereby promoting their survival. PMID- 12752218 TI - Recombinant HpaA purified from Escherichia coli has biological properties similar to those of native Helicobacter pylori HpaA. AB - The aim of this study was to recombinantly produce and purify Helicobacter pylori adhesin A (HpaA) from Escherichia coli and compare it to purified native H. pylori HpaA, for potential use as a vaccine antigen. The hpaA gene was cloned from H. pylori, transferred to two different expression vectors, and transformed into E. coli. Expression of rHpaA was analysed by immunoblot, inhibition ELISA, and semi-quantitative dot-blot. Using affinity chromatography, rHpaA was purified from E. coli and native HpaA from H. pylori. The binding of both purified proteins to sialic acid was analysed and antibody titres to native and rHpaA were compared after intraperitoneal immunisation of C57/Bl mice. The rHpaA protein was highly expressed in E. coli from both vectors. Purified recombinant and native HpaA bound similarly to fetuin but also to the non-sialylated asialofetuin. Both native HpaA and rHpaA induced comparable amounts of specific antibodies in serum after immunisation and they were identical in double immunodiffusion. In conclusion, rHpaA was successfully produced in E. coli. Purified rHpaA showed biological properties similar to those of native HpaA isolated from H. pylori and may therefore be further used as an antigen in the development of a vaccine against H. pylori infection. PMID- 12752219 TI - Routine genotyping of human papillomavirus samples in Denmark. AB - In order to examine a sensitive unbiased consensus PCR with routine sequencing for HPV typing, we analysed Danish male and female patients suspected of having an HPV infection. We used the well-characterised nested PCR setting with MY09/MY11 and GP5+/GP6+ primers, followed by routine cycle sequencing. Of 1283 clinical samples from female patients based on suspected HPV infection, we found 379 (29%) negatives and 894 (70%) positives. Samples containing >5000 HPV copies/ml were genotyped by sequencing. Of the 552 HPV genotyped samples from women >15 years of age, 398 were characterised as high-risk types and the remaining 154 as low-risk types. The most commonly found high-risk types were HPV 16, HPV-31, HPV-33, HPV-18, HPV-58, and HPV-52, and the most commonly found low risk types were HPV-6, HPV-53 and HPV-11. In addition, we observed that other typing assays could not perform as sensitively or accurately as the nested PCR/cycle sequencing method used in this study. For instance, 87 out of 552 genotyped samples could not have been typed correctly in the Hybrid Capture II assay. Of these 87 samples, 46 (53%) were considered as high-risk types. PMID- 12752220 TI - Activation of the respiratory burst by Pneumocystis carinii. Efficiency of different antibody isotypes, complement, lung surfactant protein D, and mannan binding lectin. AB - The effect of opsonization of Pneumocystis carinii with different antibody classes, complement, mannan-binding lectin (MBL), and lung surfactant protein D (SP-D) on respiratory burst activation was studied. Antibodies were obtained by affinity chromatography, complement from a hypogammaglobulinaemic patient, and phagocytic cells from blood donors. Respiratory burst activation was measured by chemiluminescence (CL). With freshly isolated neutrophils the combination of antibodies and complement but not antibody alone, had opsonizing properties. With neutrophils cultured for 20 h, however, IgG increased the CL response. In macrophages P. carinii opsonized with IgG alone induced a CL response proportional to the antibody titre used. With IgA an effect, albeit lower, was also seen, whereas IgM alone was inefficient. The combined effect of antibodies and complement increased the response significantly for all three antibody classes, IgG and complement giving the largest response. Binding of MBL to P. carinii and Candida albicans was demonstrated; however, only the former stimulated activation of the respiratory burst. SP-D did not bind to either microorganism and had no effect on the respiratory burst. It is concluded that IgG, IgA and complement are important opsonizing factors in infections involving P. carinii. The relative importance varies with the type of phagocytic cell studied. PMID- 12752221 TI - Myoepithelioma of the vulva. AB - The histologic diagnosis of myoepithelioma is often problematic. We here describe a case of myoepithelioma, composed exclusively of neoplastic myoepithelial cells, in the vulva of a 52-year-old female. The vulva is a very rare site for this neoplasm. A subcutaneous tumor measuring 3.0 x 2.5 x 2.0 cm displayed a multinodular growth pattern. Histologically, it was characterized by epithelioid, trabecular, cord, solid, or reticular arrangements of tumor cells and markedly hyalinized stroma. The tumor cells were large and polygonal with eccentrically located round to oval nuclei with prominent nucleoli and eosinophilic cytoplasm. Some cells had clear cytoplasm. Moderate cellular atypia was seen and there were 4 mitotic figures per 10 high-power fields. No ductal architecture was found and there were no areas of chondroid or osseous differentiation. There was no destructive invasive growth. Immunohistochemically the tumor was positive for vimentin, epithelial membrane antigen, wide keratin, alpha-smooth muscle actin, S 100 protein, and glial fibrillary acidic protein. The patient was well and free of disease at 6 months. This neoplasm should be distinguished from other epithelial and mesenchymal neoplasms as it shows a different clinical behavior. PMID- 12752222 TI - ICAM-1 and beta(3) integrin immunoexpression in malignant melanoma cells: can they be used as additional predictors? AB - Malignant melanoma usually progresses from the intraepidermal microenvironment through a distinct radial growth phase, in which malignant potential cannot always be accurately evaluated, to invasion of the dermis (vertical growth phase) and metastasis. During these stages malignant cells interact with each other and with the extracellular matrix. This interaction is mediated by cell surface adhesion molecules such as the beta(3) integrin subunit and ICAM-1. Our aim was to investigate whether the expression of these two molecules is associated with the various histopathologic prognosticators commonly evaluated in malignant melanoma. Using a standard three-step immunoperoxidase technique we evaluated the above molecules' expression in a documented series of 66 cutaneous malignant melanomas. Forty-five were superficial spreading melanomas, including 18 in mixed growth phase. Positive immunoreaction was estimated by image analysis. ICAM-1 immunopositivity status was significantly more frequent among malignant melanomas of the nodular type (p=0.0001), and was associated with the vertical growth phase, Breslow thickness of >0.77 mm, and with evident lymphocytic infiltration. beta(3) integrin immunopositivity showed similar results in certain respects; it was more frequently detected in superficial spreading melanomas in which vertical growth had developed (p=0.002) and in cases with regression. There appears to be an association of these molecules with histopathologic features that predict increased tumorigenicity of malignant melanocytes. PMID- 12752223 TI - Validation of the use of proliferation markers in canine neoplastic and non neoplastic tissues: comparison of KI-67 and proliferating cell nuclear antigen (PCNA) expression versus in vivo bromodeoxyuridine labelling by immunohistochemistry. AB - In order to evaluate the suitability of Ki-67 and proliferating cell nuclear antigen (PCNA) for determination of proliferative activity, the immunohistochemically determined nuclear expression of these antigens in canine non-neoplastic and neoplastic tissues was compared with the results of in vivo bromodeoxyuridine (BrdU) labelling, which - by measurement of the fraction of S phase cells - is considered as the standard in the analysis of proliferative activity. The samples investigated consisted of non-neoplastic mammary and lymphoid tissues, and of benign and malignant (primary/metastatic) mammary tumours, and malignant lymphomas. Great regional heterogeneity prevented determination of an overall labelling index (LI) in normal lymphoid tissues. In the remaining combined group of samples, LI values were significantly ranked in the order PCNA>Ki-67>BrdU. However, the correlation of Ki-67 or PCNA as compared to BrdU LI values was only moderate in the combined group [approximately 0.5, Spearman rank test] as well as in most subgroups, whilst it was very poor in the group of primary mammary cancers. These observations indicate that Ki-67 or PCNA LIs as markers of proliferation do not evenly match in vivo BrdU labelling. PMID- 12752224 TI - Clinical importance of vascular endothelial growth factor (VEGF) for papillary thyroid carcinomas. AB - Vascular endothelial growth factor (VEGF) is a major regulator of angiogenesis and may be produced by some cancer cells. Several recent reports have documented that increased expression of VEGF is associated with risk of recurrence or decreased recurrence-free survival in papillary thyroid cancers (PTC). The aims of this study were to determine whether immunohistochemical expression of VEGF is related to local and distant recurrence of PTC and to evaluate the relationship between hypervascularization and VEGF expression in papillary thyroid carcinomas. VEGF expression was examined immunohistochemically in 48 papillary carcinomas. Ten normal thyroids were used as controls. Patients were followed for 61.7 (range 24-143) months. Twelve of the patients had local and distant recurrences. VEGF immunostaining, blinded for clinicopathological data, was evaluated semiquantitatively by two pathologists. The difference between the recurrent (n:12) and nonrecurrent (n:36) carcinomas was statistically significant (p:0.001). VEGF expression was also stronger in papillary thyroid carcinomas than in normal thyroid tissues. The mean microvascular densities were significantly higher than in normal thyroid tissues. These data indicate that VEGF staining is strongly associated with increased frequency of local and distant recurrence in PTC and that the immunohistochemical profile of the expression may be used as a marker for predicting which tumors have metastatic potential. PMID- 12752225 TI - Assessment of HER-2/neu overexpression and/or gene amplification in breast carcinomas: should in situ hybridization be the method of choice? AB - AIMS: Since the release of Herceptin, pathology laboratories have been requested to test breast carcinomas for HER-2/neu overexpression and/or gene amplification. Standardized IHC and FISH are mandatory in order to get reliable results, but there are problems even with standardized procedures. We decided to evaluate the two methods to determine which, or possibly if both, should be the primary investigation method(s). METHODS AND RESULTS: The material consisted of 215 primary invasive breast carcinomas with complete clinical follow-up of 15 years. HER-2/neu protein expression was determined for all specimens, whereas FISH for assessing HER-2/neu gene signal number was done in 165 cases. IHC was double checked with two or three different antibodies in 35 tumours, including all cases with discrepancies between IHC and FISH. Among these, there were discrepancies in a third. IHC overexpression of HER-2/neu was found in 13% and gene amplification in 18%. Discordance between IHC and FISH was found in 11 cases (8%). Five tumours were IHC+/FISH- and six were IHC-/FISH+. IHC and FISH positive cases as well as FISH only positive tumours had the same prognosis respecting survival. Tumours with >2 but 4 gene signals per nucleus. In contrast, cases with IHC overexpression without gene amplification had a prognosis similar to that of IHC-/FISH- tumours. CONCLUSIONS: From our data, it seems to be more important to assess HER-2/neu gene amplification than IHC overexpression. Failure to detect FISH-amplified (IHC negative) cases would have an adverse effect on the survival of these patients. On the other hand, IHC overexpression tumours without gene amplification appear to belong to a better prognostic group, and failure to detect them would probably not have a negative effect on the survival of these women. Even though FISH is a more complex and expensive procedure, it should be considered the method of choice for primary assessment of HER-2/neu status in breast cancer patients. PMID- 12752226 TI - Association between testicular dysgenesis syndrome (TDS) and testicular neoplasia: evidence from 20 adult patients with signs of maldevelopment of the testis. AB - Based on a well established association between testicular cancer and undescended testis and more recent publications on epidemiological links between these disorders and male infertility, we proposed the existence of a testicular dysgenesis syndrome (TDS). In most cases TDS presents with impaired spermatogenesis, only in rare cases the full range of its signs, including genital malformations and testicular cancer can be seen in one patient. In order to further corroborate our hypothesis about the presence of testicular dysgenesis in patients with testicular abnormalities, we decided to re-analyse recent testicular biopsies derived from patients with infertility, hypospadias and undescended testis. We searched for histological signs of testicular dysgenesis: microliths, Sertoli-cell-only tubules, immature seminiferous tubules with undifferentiated Sertoli cells, and tubules containing carcinoma in situ (CIS) cells. We identified 20 patients who fulfilled the histological criteria for testicular dysgenesis, 9 of whom were diagnosed with uni- or bilateral testicular germ cell neoplasia, and the remaining ones with subfertility. The presence of CIS was detected in 5 patients (3 of them with overt contralateral germ cell tumours). In all but one of the CIS cases, at least one additional sign of testicular dysgenesis was detected. Clinical records of all patients were subsequently analysed. The majority of cases had oligozoospermia or azoospermia. Their reproductive hormone profiles correlated with the results of semen sampling and testicular histology. In conclusion, our study of 20 patients with various reproductive abnormalities provided evidence that TDS is a real clinical entity. We speculate that most of these abnormalities are caused by adverse environmental effects rather than specific gene mutations. PMID- 12752228 TI - Multinucleated spermatogonia in cryptorchid boys: a possible association with an increased risk of testicular malignancy later in life? AB - At birth, undescended testes contain germ cells, but after 1 year of life, a reduced number of germ cells is generally found. Microlithiasis and carcinoma-in situ-testis occur in cryptorchid boys. Multinucleated germ cells, including at least 3 nuclei in the cell, exist in impaired spermatogenesis and in the senescent testis. AIM OF THE STUDY: We investigated whether multinucleated spermatogonia were present in undescended testes of cryptorchid boys, and if such a pattern is associated with special clinical features. RESULTS: Multinucleated spermatogonia occurred in 13/168 (8%) of 163 consecutive cryptorchid boys, who underwent surgery for cryptorchidism with simultaneous testicular biopsy showing seminiferous tubules. The patients with multinucleated spermatogonia more often exhibited a normal germ cell number (Fisher's exact test, p<0.0005), and were younger at surgery (Mann Whitney, p<0.005) than the rest of the patients. Before surgery, 3 patients underwent treatment with Erythropoietin because of renal failure. An intra-abdominal testis underwent clipping and division of the spermatic vessels, and a biopsy at final surgery 7 months later, exhibited multinucleated spermatogonia. In 1 case the undescended testicular position, a fixed retraction, was acquired after surgery for an inguinal hernia. Multinucleated spermatogonia were found in cases of carcinoma-in situ-testis in 2 cryptorchid boys. No case of multinucleated germ cells appeared in our normal material. CONCLUSION: Multinucleated spermatogonia are a further abnormality present in cryptorchidism. The cryptorchid boys with multinucleated spermatogonia in general exhibited rather many germ cells. This feature may be associated with an increased risk of testicular malignancy later in life, and we propose a careful follow up regime in these cases including ultrasound examination and a testicular biopsy in cases of symptoms or clinical findings. PMID- 12752231 TI - Morphology of testicular parenchyma adjacent to germ cell tumours. An interim report. AB - A comparative morphological analysis of parenchyma adjacent to testicular germ cell tumours (TGCT) was performed in a series of 181 orchidectomy specimens: 86 with seminomas (Se), 72 with nonseminomatous germ cell tumours (NS) and 23 with combined tumours (CT, which have a Se and a NS component). The following morphological features were semiquantitatively scored: spermatogenesis (modified Johnsen score); amount of tubular atrophy; amount of carcinoma in situ (CIS); amount of intertubular tissue. Absence and presence was scored for the following features: lymphocytic infiltrate surrounding and invading CIS; intratubular seminoma (ISe); intratubular nonseminoma (INS); microlithiasis; diffuse and nodular hyperplasia of Leydig cells; angioinvasiveness; testicular angiopathy. Using non-parametric statistics these features were correlated with each other and with tumour type, tumour size and age of the patient. Se-patients presented at significantly higher age than NS-patients (36 vs 29 years, p=0.001). The age of patients with CT (32 years) was in between that of Se- and NS-patients. No correlation was found between patient age and tumour size. Parenchyma adjacent to Se, compared to parenchyma adjacent to NS had the following significant differences: a lower Johnsen score (5.6 vs 7.2, p=0.005); less frequent (85% vs 97% of specimens, p=0.016) and a lesser amount of CIS (26% vs 32% of tubules, p=0.015); more frequent peri- (80% vs 60% of specimens, p=0.001) and intratubular (68% vs 30% of specimens, p=0.001) lymphocytic infiltrates; more extensive tubular atrophy (36% vs 15% of tubules, p=0.001); and a larger area of intertubular tissue (42% vs 34% of parenchyma area, p=0.016). The pooled Se and CT had a significantly higher frequency of ISe than the NS (31% vs 17% of specimens, p=0.036). With one exception INS was only found adjacent to NS or CT, with a frequency of 16%, and 20% of the specimens, respectively. It was significantly associated with angio-invasiveness. In specimens lacking angio invasion the frequency of INS was 6%. The correlation of INS with tumour size and patient age was studied in a series of 145 NS and CT (95 from the original series supplemented by 50 newer cases). In this series INS was significantly associated with smaller tumours and younger patients. Extensive tubular atrophy was significantly correlated with higher age, the diagnosis of Se, a low Johnsen score, and the presence of angiopathy. The more tubular atrophy, the less CIS (both in incidence and amount). Inversely, a higher Johnsen score is associated with smaller tumours, the diagnosis of NS or CT, a higher incidence and a larger amount of CIS, and little tubular atrophy. Tubules with mature spermatogenesis were found in 42% of the specimens regardless of tumour type. We conclude that ISe and INS are probably frequent intermediate stages between CIS and Se and NS, respectively. The features of parenchyma adjacent to Se are probably due to the host response elicited by the invasive Se, which secondarily also affects CIS. The long time to clinical presentation allows the host to eradicate most of the CIS by the time the tumour is surgically removed. The much less extensive morphological features of a host response in parenchyma adjacent to NS support the contention that NS originates as INS, behind the blood/testis barrier, without exposure of the host to tumour cells with a seminomatous phenotype (CIS- or Se cells). Microlithiasis and testicular angiopathy are frequent, but not specific findings in parenchyma next to TGCT. Their relationship with the development with TGCT is unexplained. PMID- 12752232 TI - Epidemiology of gonadal germ cell cancer in males and females. AB - The epidemiology of testicular germ cell cancer is relatively well understood, but less is known about the epidemiology of ovarian germ cell cancer. Cases of testicular (7910) and ovarian (453) germ cell cancer diagnosed 1960-1999 were extracted from the Thames Cancer Registry. The incidence rate in males doubled in the period from 2.0 to 4.4 per 100,000. In females the rate was much lower but the rate of increase over the period was similar to the increase in males. In both sexes the incidence increased sharply around the age of onset of puberty and decreased in older age groups. The biological mechanism of initiation of germ cell carcinogenesis is similar in males and females and probably occurs with roughly the same frequency in the two sexes. In males, the initiated cell population is promoted by the normal action of adult sex hormones to form carcinoma in situ, consisting of many millions of cells. The incidence of testicular cancer is rate limited by the development of carcinoma in situ. The key difference between males and females, which may explain the different levels of incidence, is the much lower number of susceptible cells in females at the time of puberty. PMID- 12752236 TI - Diagnosis and management of testicular intraepithelial neoplasia (carcinoma in situ)--surgical aspects. AB - Germ cell tumours (CT) are no true carcinomas; therefore the term testicular intraepithelial neoplasia (IN) is probably more appropriate than "CIS". The diagnostic accuracy of a single-site biopsy is an open question. We experienced 9 false-negative biopsies among 1859 cases. Thus, the proportion of a failed diagnosis is 0.5%. The main reason for diagnostic failure is the non-random distribution of TIN within the testicle. Currently we are investigating whether a two-site biopsy is more accurate than a single biopsy. In the ongoing trial, the over-all prevalence of TIN is around 5.3%, so far. In one quarter of the positive cases the lesion was found in only one of the two specimens. Thus, a double biopsy appears to be more favourable than the traditional single biopsy. Surgical complications amount to 2.5% in that double biopsy study. Only one surgical re intervention was required among 983 patients. Serial imaging studies with scrotal sonography and magnetic resonance imaging (MRI) disclosed a transient intratesticular haematoma/oedema postoperatively. So, testicular biopsy, even when performed at two sites is in fact a low-complication procedure. Low dose radiotherapy to the testis is the treatment of choice for TIN. However, more than one quarter of patients require testosterone supplementation secondary to androgen-deficiency. Two dose-reduction studies (Denmark and Germany) had to be terminated prematurely because unexpected relapse of TIN was encountered at 14 Gy and 16 Gy. Possibly, hyperfragmentation schedules can overcome the antagonism of androgenic compromise and oncological safety. In a nation-wide survey, it was shown that contralateral biopsies were routinely performed in 66% of the urological departments in Germany. Another 19% offered the biopsy to particular "risk-cases"; only 15% never did a biopsy. Among those refusing biopsies, there was a higher proportion of small hospitals and a significantly lower annual case number of GCT, when compared to those doing the biopsy. Thus, the contralateral biopsy is a well-established procedure among German urologists; those with a high caseload of GCT particularly appreciate it. PMID- 12752235 TI - Extragonadal germ cell tumors: relation to testicular neoplasia and management options. AB - An unselected population of 635 consecutive extragonadal GCT patients (EGCT) treated between 1975 through 1996 at 11 cancer centers was retrospectively evaluated for clinical prognosis and biological features of this disease. Five hundred twenty-four patients (83%) had a nonseminomatous GCT, and 104 patients (16%) a seminomatous histology; 341 (54%) patients had a primary mediastinal EGCT, and 283 patients (45%) a retroperitoneal EGCT. Following platinum based induction chemotherapy+/-secondary surgery, 141 patients (49%) with mediastinal nonseminomas (median follow up period: 19 months) and 144 patients (63%) with retroperitoneal nonseminoma (median follow up period: 29 months) are alive [p=0.0006]. In contrast, the overall survival rate for patients with seminomatous EGCT is 88% with no difference between patients with mediastinal or retroperitoneal tumor location (median follow up period: 49 months). Multivariate analysis revealed nonseminomatous histology, the presence of non-pulmonary visceral metastases, primary mediastinal GCT location, and elevated beta-HCG as independent prognostic factors for shorter survival. Sixteen patients (4.1%) developed a metachronous testicular cancer despite the use of platinum based chemotherapy. The cumulative risk of developing a MTC 10-years after a diagnosis of EGCT was 10.3% (95% CI=4.9 to 15.6%), but higher among patients with nonseminomatous EGCT (14.3%; 95% CI=6.7 to 21.9%) or retroperitoneal EGCT location (14.2%; 95% CI=5.6 to 22.8%) than among patients with seminomatous EGCT (1.4%; 95% CI=0.0 to 4.2) or mediastinal EGCT location (6.2%; 95% CI=0.1 to 12.2). After a median follow-up of 51 months (range=1 to 154 months), all 16 MTC patients were alive without disease. Patients with pure seminomatous EGCT histology have a long term chance of cure of almost 90% irrespective of the primary tumor site. Patients with mediastinal nonseminomas have a five-years survival rate of 45%. This outcome is clearly inferior compared to patients with nonseminomatous retroperitoneal primaries who have a five-year survival rate of 62%. PMID- 12752238 TI - Update on the diagnostic safety for detection of testicular intraepithelial neoplasia (TIN). AB - Testicular intraepithelial neoplasia (TIN) of the testis is the noninvasive precursor of testicular germ cell tumours (GCT) and can be detected by a single random biopsy in 5% of patients with GCT in the contralateral testes. Although it is generally presumed that TIN is dispersed throughout the testis, we realize in about 60% of TIN bearing tissue close to testis tumours that its distribution is not homogenously diffuse, but may be focal. Thus we tested whether we can improve diagnostic safety in detecting TIN by increasing the number of biopsies. We could finally evaluate 295 men with proven testicular tumours. Three biopsies of contralateral testes were taken (each 5 mm length) from one surgical incision site and fixed in Bouin's solution or glutaraldehyde. TIN cells were histologically identified by their typical morphological characteristics and additionally by placental alkaline phophatase (PlAP) immunohistochemistry. Patients revealed testicular tumour without contralateral TIN in 271 cases and with contralateral TIN in 24 cases (8.1%). In 6 of these 24 men with contralateral TIN the cells could be detected in only one (n=5) or two (n=1) of the three specimen investigated. That means in these six patients TIN could have been missed if only one single random biopsy was taken. By increasing the number of biopsies (=increasing the number of investigated seminiferous tubules) the detection rate of contralateral TIN may be increased up to 8.1%. Thus we recommend multiple testicular biopsies to increase the diagnostic safety in detection of TIN. Biopsies may be taken from one randomly chosen surgical incision site. PMID- 12752240 TI - Surveillance in stage I testicular cancer. AB - Treatment results on 695 stage I testicular cancer patients followed with surveillance are described. Seminoma (SGCT) was present in 394 patients and nonseminoma (NSGCT) in 301 patients. Relapses were detected in 155 patients (22%), in 69 patients with SGCT (17%) and 86 with NSGCT (29%). In patients with vascular invasion, relapse was detected in 54% of patients with NSGCT and 38% of patients with SGCT. Time to relapse was median 13 months (range 1 to 84 months) for SGCT and 5 months (range 1 to 171 months) for NSGCT. Forty-nine percent of relapses in SGCT patients were seen within the first year, 87% of the relapses were diagnosed within the first two years, and 98% of the relapses were detected within 5 years. The figures for NSGCT were 80%, 89% and 95%, respectively. Forty five patients had carcinoma in situ in the contralateral testis, 62% had this together with a seminoma in the other testis. Ten patients died during the follow up period. None of these deaths were caused by the germ cell tumour or the treatment. The overall survival for patients with stage I disease is 98.6%, and the cause specific survival 100%. PMID- 12752243 TI - Testis conserving chemotherapy in germ cell cancer: its potential to increase understanding of the biology and treatment of carcinoma-in-situ. AB - Prompted by recognition of the potential of chemotherapy to increase the success of testis conserving surgery in patients with germ cell cancer, background and outcome data are reviewed and their contribution to the ongoing debate about how germ cell cancer develops discussed. The review is based on three previous studies of: a) time trends in tumour size in 578 personal series of all stages of testis cancer treated since 1978; b) impact of chemotherapy on actuarial risk of tumours in contralateral testis examined on 1221 patients treated in trials through the Anglian Germ Cell Cancer Consortium; and c) testes conservation attempted using chemotherapy in 78 patients. Since 1978 tumour size has decreased from 4.8 to 3.0 cms while cure has gone from 77 to 97%. There was no overall long term reduction in second cancers beyond 10 years in stage 1 patients after orchidectomy alone compared to stage 1 or metastatic disease patients receiving chemotherapy though the incidence was non significantly lower up to 10 years particularly in those patients receiving etoposide based combination. Testis conservation was initially successful in 28 of 78 (36%). An additional 25 (32%) had no viable cancer in orchidectomy specimen. In the 28 primary tumours cured by chemotherapy there was a 26% late relapse rate between 5 and 10 years (all cured by orchidectomy) compared to less than 5% in those cured with established metastases. In conclusion, testis conservation with chemotherapy is safe and feasible, though relapse is too frequent for routine service use. Confirmation of the high frequency of late relapse by others has raised the question whether these recurrences are due to post pubertal events reinducing CIS in intrauterine oestrogen primed germ cells and highlights the potential of testes conservation studies to better understand germ cell cancer development. PMID- 12752244 TI - Endocrine function in patients treated for carcinoma in situ in the testis with irradiation. AB - CIS is found in the contralateral testis in 5% of the patients with testicular germ cell cancer. The management of CIS in the contralateral testis is important because the majority - if not all - cases of CIS will progress to invasive disease without treatment. It is well documented that testicular irradiation with a total dose of 14-20 Gy (2 Gy x 7-10) is an effective and safe treatment for CIS in the contralateral testis in patients with unilateral testicular germ cell cancer. However few relapses of testicular cancer have been observed in testis treated with these regimens and the data on 14 Gy are sparse. One study has indicated that more radiotherapy with lower doses per fraction could be useful, but more data are needed to confirm this. Endocrine testicular function has been shown to be impaired already before treatment in patients with CIS and is further impaired after testicular irradiation with 14-20 Gy (2 Gy x 7-10) and only minor dose dependency is seen in the impairment of Leydig cell function. The optimal treatment of CIS in the contralateral testicle in patients orchidectomised for testicular cancer seems to be local radiotherapy of the testis with CIS in order to preserve at least a part of the Leydig cell function. However, the optimal dose level has to be defined. PMID- 12752246 TI - A new prognostic model for testicular germ cell tumours. AB - In univariate analyses of patients with metastatic testicular germ cell tumours (TGCT), both the International Germ Cell Consensus Classification (IGCCC) and serum lactate dehydrogenase (S-LD) isoenzyme 1 catalytic concentration (S-LD-1) significantly predicted survival. In complementary analyses of 81 patients with metastatic TGCT, S-LD and S-LD-1 classified the prognosis differently for 23 patients. In multivariate Cox hazard analyses of risk factors, only IGCCC and S LD-1 predicted the prognosis (p=0.036, and p=0.0007, respectively). A new prognostic model based on prognostic information from main histology, IGCCC, and S-LD-1 changed the prognostic prediction by IGCCC for 19 patients (24%). Judged by to the area under the curve for receiver operation characteristics curves, the new model predicted five-years survival for the patients better than IGCCC and a modified version of the third edition of the TNM classification (p=0.025, and p=0.01, respectively). However, new studies should validate the new model before it is recommended as a general classification system of patients with metastatic TGCT. PMID- 12752248 TI - Y chromosome haplogroups: a correlation with testicular dysgenesis syndrome? AB - Testicular dysgenesis syndrome encompasses low sperm quality, hypospadias, cryptorchidism and testicular cancer. Epidemiological studies and genetic data from familial cases suggest that testicular dysgenesis syndrome has a common etiology. The Y chromosome is known to encode genes that are involved in germ cell development or maintenance. We have therefore investigated if different classes of Y chromosomes in the general population (Y chromosome haplogroups) are associated with aspects of the testicular dysgenesis syndrome. We defined the Y chromosome haplogroups in individuals from different European counties who presented with either (i) oligo- or azoospermia associated with a Y chromosome microdeletion, (ii) unexplained reduced sperm counts (<20 x 10(6)/ml) or (iii) testicular cancer. We failed to find Y chromosome haplotype associations with either microdeletion formation or testicular cancer. However, in a study of the Danish population, we found that a specific Y chromosome haplogroup (hg26) is significantly overrepresented in men with unexplained reduced sperm counts compared with a Danish control population. The factors encoded by genes on this class of Y chromosome may be particularly susceptible to environmental influences that cause testicular dysgenesis syndrome. Our current data highlight the need for further analyses of clinically well-defined patient groups from a wide range of ethnic and geographic origins. PMID- 12752250 TI - Polymorphic DAZ gene family in polymorphic structure of AZFc locus: Artwork or functional for human spermatogenesis? AB - Human spermatogenesis is regulated by a network of genes located on autosomes and on sex chromosomes, but especially on the Y chromosome. Most results concerning the germ cell function of the Y genes were obtained by genomic breakpoint mapping studies of the Y chromosome of infertile patients. Although this approach has the benefit of focussing on those Y regions that contain most likely the Y genes of functional importance, its major drawback is the fact that fertile control samples were often missing. In fertile men, molecular and cytogenetic analyses of the Y chromosome has revealed highly polymorphic chromatin domains especially in the distal euchromatic part (Yq11.23) and in the heterochromatic part (Yq12) of the long arm. In sterile patients cytogenetic analyses mapped microscopically visible Y deletions and rearrangements in the same polymorphic Y regions. The presence of a Y chromosomal spermatogenesis locus was postulated to be located in Yq11.23 and designated as AZoospermia Factor (ZF). More recently, molecular deletion mapping in Yq11 has revealed a series of microdeletions that could be mapped to one of three different AZF loci: AZFa in proximal Yq11 (Yq11.21), AZFb and AZFc in two non-overlapping Y-regions in distal Yq11 (Yq11.23). This view was supported by the observation that AZFa and AZFb microdeletions were associated with a specific pathology in the patients' testis tissue. Only AZFc deletions were associated with a variable testicular pathology and in rare cases AZFc deletions were even found inherited from father to son. However, AZFc deletions were found with a frequency of 10-20% only in infertile men and most of them were proved to be "de novo", i.e. the AZFc deletion was restricted to the patient's Y chromosome. Based mainly on positional cloning experiments of testis cDNA clones and on the Y chromosomal sequence now published in GenBank, a first blueprint for the putative gene content of the AZFc locus can now be given and the gene location compared to the polymorphic DNA domains. This artwork of repetitive sequence blocks called AZFc amplicons raised the question whether the AZFc chromatin is still part of the heterochromatic domain of the Y long arm well known for its polymorphic extensions or is decondensed and part of the Yq11.23 euchromatin? We discuss also the polymorphic DAZ gene family and disclose putative origins of its molecular heterogeneity in fertile and infertile men recently identified by the analyses of Single Nucleotide Variants (SNVs) in this AZFc gene locus. PMID- 12752252 TI - Localisation of susceptibility genes for familial testicular germ cell tumour. AB - Approximately 1700 men in the United Kingdom develop testicular germ cell tumours (TGCT) per year. Among the known risk factors a family history of disease remains one of the strongest (1, 2). Two-percent of TGCT cases report another affected family member. Epidemiological studies have shown that there is an eight to ten fold increase in relative risk of TGCT to brothers of patients and a fourfold increased risk to fathers and sons (2-5). This relative risk is considerably higher than for most other common cancers, which rarely exceeds four and strongly suggests that genes may play an important role in TGCT. Linkage analysis of the set of families compatible with X-linkage (i.e. no male to male transmission) provided the first statistically significant evidence for a TGCT predisposition locus (6). The gene called TGCT1 is located at Xq27 and seems to be associated with a risk of bilateral disease and undescended testis. However TGCT1 does not account for all TGCT pedigrees and additional susceptibility genes must exist. Our group has now genotyped 179 TGCT pedigrees and identified additional genomic regions that might also harbour TGCT susceptibility genes. This paper reviews the current data for the region at Xq27 and presents evidence for several other possible candidate regions. PMID- 12752254 TI - The testicular germ cell tumour genome. AB - Human testicular germ cell tumour (TGCT) of adolescents and young adults develop from precursor lesions called carcinoma in situ (CIS), which is believed to originate from diploid primordial germ cells during foetal life. CIS is initiated by an aneuploidisation event accompanied by extensive chromosome instability. The further transformation of CIS into invasive TGCT (seminomas and nonseminomas) is associated with increased copy number of chromosome arm 12p, most often seen as isochromosome 12p. Despite the morphological distinctions between seminomatous and nonseminomatous TGCTs, they have many of the same regional genomic disruptions, although frequencies may vary. However, the two histological subtypes have quite distinct epigenomes, which is further evident from their different gene expression patterns. CIS develops from cells with erased parental imprinting, and the seminoma genome is under-methylated compared to that of the nonseminoma genome. High throughput microarray technologies have already pinpointed several genes important to TGCT, and will further unravel secrets of how specific genes and pathways are regulated and deregulated throughout the different stages of TGCT tumourigenesis. In addition to acquiring new insights into the molecular mechanisms of TGCT development, understanding the TGCT genome will also provide clues to the genetics of human embryonic development and of chemotherapy response, as TGCT is a good model system to both. PMID- 12752256 TI - Germ cell tumours in neonates and infants: a distinct subgroup? AB - Human germ cell tumours (GCTs) constitute a heterogeneous group of tumours that can be classified into four major subgroups. One of these subgroups encompasses (immature) teratomas and yolk sac tumours of patients under the age of 5 years. In this paper we review the various clinical, histological and cytogenetical aspects of these infantile GCTs. The primordial germ cell (PGC) has been suggested to be the cell of origin for GCTs. Infantile GCTs, however, have been suggested to originate from PGCs at a different stage of maturation than adult GCTs. The cytogenetic constitution of infantile GCTs also appears to differ from the adult GCTs and includes recurrent losses of lp and 6q. Recently, two cases of infantile GCT were detected with constitutional 12q13 translocations. These exceptional cases may be instrumental in the search for candidate genes related to infantile and/or adult GCT development. PMID- 12752258 TI - Role of gain of 12p in germ cell tumour development. AB - Within the human testis, three entities of germ cell tumours are distinguished: the teratomas and yolk sac tumors of newborn and infants, the seminomas and nonseminomas of adolescents and young adults, referred to as testicular germ cell tumours (TGCT), and the spermatocytic seminomas. Characteristic chromosomal anomalies have been reported for each group, supporting their distinct pathogenesis. TGCT are the most common cancer in young adult men. The initiating pathogenetic event of these tumours occurs during embryonal development, affecting a primordial germ cell or gonocyte. Despite this intra-uterine initiation, the tumour will only be clinically manifest after puberty, with carcinoma in situ (IS) as the precursor. All invasive TGCT, both seminomas and nonseminomas, as well as CIS cells are aneuploid. The only consistent (structural) chromosomal abnormalities in invasive TGCT are gains of the short arm of chromosome 12, mostly due to isochromosome (i(12p)) formation. This suggests that an increase in copy number of a gene(s) on 12p is associated with the development of a clinically manifest TGCT. Despite the numerous (positional) candidate gene approaches that have been undertaken thus far, identification of a causative gene(s) has been hampered by the fact that most 12p gains involve rather large genomic intervals, containing unmanageable numbers of candidate genes. Several years ago, we initiated a search for 12p candidate genes using TGCT with a restricted 12p-amplification, cytogenetically identified as 12p11.2 p12.1. This approach is mainly based on identification of candidate genes mapped within the shortest region of overlap of amplification (SROA). In this review, data will be presented, which support the model that gain of 12p-sequences is associated with suppression of apoptosis and Sertoli cell-independence of CIS cells. So far, DAD-R is one of the most likely candidate genes involved in this process, possibly via N-glycosylation. Preliminary results on high through-put DNA- and cDNA array analyses of 12p-sequences will be presented. PMID- 12752260 TI - Genetic control of susceptibility to spontaneous testicular germ cell tumors in mice. AB - Testicular germ cell tumors (TGCTs) are the most common cancer affecting young men. TGCT is a polygenic trait and genes that control susceptibility for TGCT development have not yet been identified. The 129/Sv inbred strain of mice is an important experimental model to study the genetics and development of TGCTs. We review several novel approaches that were developed to study the susceptibility of TGCTs in the 129/Sv mouse model and its application in humans. These approaches showed that several spontaneous and engineered mutations interact with 129/Sv-derived susceptibility genes to enhance or suppress susceptibility; two of these mutations (Ter and Trp53) revealed novel linkages for susceptibility genes in sensitized polygenic trait analysis. Linkage analysis with a chromosome substitution strains suggests that as many as 100 genes control susceptibility. Bilateral TGCTs result from the coincidental occurrence of unilateral tumors. These results highlight the important contributions that this mouse model can make to studies of TGCT susceptibility in humans. PMID- 12752262 TI - GDNF-induced seminomatous tumours in mouse--an experimental model for human seminomas? AB - Glial-cell-line-derived neurotrophic factor (GDNF) is a distant member of the transforming growth factor superfamily. It binds to and activates a receptor complex consisting of GFR-alpha1 and Ret receptor tyrosine kinase. In testis, GDNF is expressed by Sertoli cells. We have shown by transgenic loss- and gain-of function mouse models that GDNF regulates the cell fate decision of undifferentiated spermatogonia. In the GDNF +/- mice, the spermatogonia differentiate in excess leading to the depletion of germ cells. In the mice overexpressing GDNF in testes, undifferentiated spermatogonia accumulate in the tubules, no sperm is produced, and the mice are infertile. After a year, the GDNF overexpressing mice frequently (89%) develop testicular tumours, and most of them are bilateral (56%). All these tumours show the same histological pattern. They are composed of round spermatogonial/gonocytic cells with only a scant cytoplasm. The tumours are locally invasive but do not metastasise. They express germ line markers, are positive for alkaline phosphatase, and aneuploid with a triploid peak. Thus, by several histological, molecular, and histochemical characteristics, the GDNF-induced tumours mimic classical seminomas in men, but the precursor lesions are apparently different in mouse and man. PMID- 12752266 TI - Human tissue kallikreins and testicular cancer. AB - Human tissue kallikreins are fifteen homologous genes encoding for secreted serine proteases and residing tandemly on chromosome 19q13.4. These enzymes are highly expressed in a variety of tissues and participate in diverse physiological processes. Human tissue kallikreins were found to be associated with several malignancies, especially endocrine-related cancers, including prostate, ovarian, breast and testicular cancer. In testicular germ cell tumors, some tissue kallikrein genes, including KLK5, KLK10, KLK13 and KLK14, were found to be significantly down-regulated. Tissue-specific splice variant forms of some kallikreins have been identified in the testis. In this paper, the expression of KLK5, KLK10, KLK13 and KLK14 in testicular cancer and their possible roles during testicular cancer development, as well as their clinical applications are briefly reviewed. PMID- 12752264 TI - Growth regulatory factors and signalling proteins in testicular germ cell tumours. AB - The molecular basis of testicular germ cell tumourigenesis are not well elucidated. Growth factors regulate cell growth, differentiation and apoptosis. Major families of growth factors are present in the male gonad from early fetal development to adult life. They are involved in germ cell proliferation and differentiation. Growth signalling pathways suffer deregulation in many human malignancies. Given the importance of growth signals in normal testicular development and their acquired deregulation in most human cancers, growth factors and signalling molecules that have been implicated in the genesis of testicular germ cell tumours, are reviewed. We detected a somatic mutation of SMAD4 gene, responsible for loss of protein function in seminomas. This mutational inactivation may affect the activity of several members of TGFbeta superfamily (TGFbeta, activin, inhibin, BMP). VEGF expression has been shown to predict metastasis in seminomas. A significant association of HST-1 expression, a member of fibroblast growth factors, with the nonseminomatous phenotype and with tumour stage has been described. In contrast, C-KIT is expressed by seminomas only, from the preinvasive stage. Despite intense expression in almost all seminomas, activating mutation of C-KIT gene is seldom reported. Recently, the first animal model of classical testicular seminoma has been identified in transgenic mouse overexpressing GDNF. RET (GDNF receptor) expression is demonstrated in human seminomas, and not in nonseminomatous tumours. However, the exact molecular alterations of GDNF/RET/GFRalpha1 complex in germ cell tumours are not known. Finally, beside growth factors, other signalling molecules such as peptide hormones may be involved in testicular carcinogenesis. We have demonstrated a specific pattern of somatostatin receptors expression in each type of testicular germ cell tumours, with a loss of sst3 and sst4 in seminomas and loss of sst4 and expression of sst1 in nonseminomas only. These data suggest an antiproliferative action of somatostatin in testicular cancers. In summary, many growth factors and signalling molecules seem to represent specific markers for different histological types of germ cell tumours (seminomas versus nonseminomas) and may play a role in the differentiation of germ cell tumours. Despite a complex signalling pathway involved in the physiological functions of male gonad, little is known about the implication of this signalling network in testicular malignancies. From a practical stand-point, further studies on the role of growth factors in human germ cell tumours may offer a new therapeutical perspective with the development of specific pharmacological signalling modulators that could be used as therapeutic agents. PMID- 12752269 TI - Angiotensin I-converting enzyme and potential substrates in human testis and testicular tumours. AB - The angiotensin I-converting enzyme (ACE, kininase II, CD143) shows a broad specificity for various oligopeptides. Besides the well-known conversion of angiotensin I to II, ACE degrades efficiently kinins and the tetrapeptide AcSDKP (goralatide) and thus equally participates in the renin-angiotensin system, the kallikrein-kinin system, and the regulation of stem cell proliferation. In the mammalian testis, ACE occurs in two isoforms. The testicular isoform (tACE) is exclusively expressed during spermatogenesis and is generally thought to represent the germ cell-specific isozyme. However, we have previously demonstrated that, in addition to tACE, the somatic isoform (sACE) is also present in human germ cells. Similar to other oncofoetal markers, sACE exhibits a transient expression during foetal germ cell development and appears to be a constant feature of intratubular germ cell neoplasm, the so-called carcinoma-in situ (CIS) and, in particular, of classic seminoma. This demands the existence of specific paracrine functions during male germ cell differentiation and development of male germ cell tumours, which are mediated by either of the two ACE isoforms. Considering the complexity of current data about ACE, a logical connection is required between (I) the precise localisation of ACE isoforms, (I) the local access to potential substrates and (II) functional data obtained by knockout mice models. The present article summarises the current knowledge about ACE and its potential substrates with special emphasis on the differentiation restricted ACE expression during human spermatogenesis and prespermatogenesis, the latter being closely linked to the pathogenesis of human germ cell tumours. PMID- 12752270 TI - The role of retinoblastoma protein family in the control of germ cell proliferation, differentiation and survival. AB - Retinoblastoma family proteins pRb, p107 and p130 are differentially expressed in the rat testis. They function in specific cell types during testicular development and spermatogenesis, participating in the control of proliferation, differentiation, and survival. Their expression levels and phosphorylation status are modulated during germ cell cycle progression and apoptosis. Hyperphosphorylated states and elevated levels of p107 are correlated with cell cycle progression, whereas hypophosphorylated states and reduced levels are associated with suppression of proliferation and apoptosis in germ cells and Leydig cells. These proteins may also serve as markers of cell cycle status of germ cells during spermatogenesis. PMID- 12752274 TI - Current views on the pathogenesis of testicular germ cell tumours and perspectives for future research: highlights of the 5th Copenhagen Workshop on Carcinoma in situ and Cancer of the Testis. AB - This review article highlights the most important contributions presented at the 5th Copenhagen Workshop on Carcinoma in situ and Cancer of the Testis, which was held in Denmark, August 29-31, 2002. The major themes that emerged at the meeting are critically discussed and perspectives for future research in this field are presented. PMID- 12752272 TI - The emerging phenotype of the testicular carcinoma in situ germ cell. AB - This review summarises the existing knowledge on the phenotype of the carcinoma in situ (CIS) cell. CIS is a common pre-invasive precursor of testicular germ cell tumours of adolescents and young adults. These tumours display a variety of histological forms. Classical seminoma proliferates along the germ cell lineage, whereas embryonal carcinoma retains embryonic features and readily differentiates into teratomas that resemble various somatic cell lineages. A thorough review of the gene expression in CIS cells in comparison to normal testicular germ cells and overt tumours supports the view that CIS is a common precursor for both tumour types. Impaired cell differentiation resulting in a partial retention of the embryonic features, associated with an increasing genomic instability may be responsible for a remarkable phenotypic heterogeneity of CIS cells. Depending on the degree of differentiation and pluripotency, CIS cells found in adult patients seem to be predestined for further malignant progression into one or the other of the two main types of overt tumours. A new concept of phenotypic continuity of differentiation of germ cells along germinal lineage with a gradual loss of embryonic features based on the analysis of gene expression in all types of germ cells during their ontogeny is presented in this review. The data point out that despite the phenotypic continuum of gene expression, there are two periods of rapid changes of gene expression: first at the transition from primordial germ cells to pre-spermatogonia, and later during the pubertal switch from the mitotic to meiotic cell division. The persistent expression of embryonic traits in CIS cells, and the high expression of the cell cycle regulators that are typical of mitotic germ cells support our long-standing hypothesis that CIS cells originate from primordial germ cells or gonocytes and not from germ cells in the adult testis. PMID- 12752275 TI - Testicular Germ Cell Neoplasia: A Developmental Disease? Proceedings of the 5th Copenhagen Workshop on Carcinoma in situ and Cancer of the Testis: Environmental and Genetic Aspects. Copenhagen, Denmark, August 29-31, 2002. PMID- 12752277 TI - The keystone design perforator island flap in reconstructive surgery. PMID- 12752278 TI - Role of pelvic lymphadenectomy as an adjunct to radical prostatectomy. PMID- 12752279 TI - Intraoperative lymph node staging of prostate cancer: the case against. AB - BACKGROUND: Intraoperative frozen section analysis of obturator nodes is an accepted screening procedure, excluding from prostatectomy that group of node positive patients who are presumed to almost always have disseminated cancer. The overall efficacy of this procedure depends not only on the near inevitability of cancer progression in these patients, but also the procedure morbidity (previously estimated at 8.4%), the additional costs (currently estimated at A$1200) and the infrequency of positive nodes. We evaluate the efficacy of lymph node staging for prostate cancer. METHODS: We have evaluated the efficacy of intraoperative screening by node dissection in 123 prostatectomy cases. These cases were prescreened from a series of 261 radical prostatectomies by evaluating preoperative serum PSA and Gleason grade. RESULTS: Three patients were identified with nodal disease, representing a detection rate of 2.4%. The present study confirms that current trends in prostate cancer identification and selection of individuals for radical surgery very rarely identify node-positive disease even after preselection with accepted 'high-risk' markers. CONCLUSION: Considering the attendant cost and morbidity, there appears to be no justification for lymph node dissection as a routine preliminary to prostatectomy. PMID- 12752280 TI - Intermediate to long-term results of repairing incompetent multiple deep venous valves using external valvular stenting. AB - INTRODUCTION: Chronic venous insufficiency is a major cause of morbidity in the community. The purpose of the present study was to assess the efficacy and safety of repairing multiple deep venous valves with an External Valvular Stent. METHODS: Forty-two limbs with chronic venous disease were operated on between 1987 and 1991. The patients included in the present series have now had more than 5 years of follow up (mean: 7.9 years; range: 5.4-11.9 years). Most patients had primary deep venous incompetence. There were 125 valve repairs out of 146 venographically suitable valves primarily in the superficial femoral and popliteal veins (2.98 per limb). Follow up was incomplete because of the duration of the trial. RESULTS: Following surgery ulcer areas decreased from a mean preoperative value of 12.9 cm2 to 1.2 cm2 at 86 months with almost 80% of the ulcers healed. All symptoms, pain, swelling, cramps and pigmentation were improved at a statistically significant level. The 90% infrared photoplethysmography recovery time almost doubled from 6.7 s preoperatively to 12.4 s at 36 months and were unchanged in the long term. Venous pressures were only significant at 12 months post implantation. Using multivariant analysis, the number of stents implanted was statistically associated with an increased number of ulcers healing. The site of venous valve repairs seems to be irrelevant to all of the variables. CONCLUSION: Multiple deep venous valve repairs are appropriate and the best form of treatment for specifically selected individuals with primary deep venous incompetence. PMID- 12752281 TI - Acute generalized peritonitis in African children: assessment of severity of illness using modified APACHE II score. AB - BACKGROUND: Generalized surgical acute abdomen is a significant cause of morbidity and mortality among children. Severity assessment is useful in order to prioritize treatment and reduce complications. Patients with a high severity score are often faced with high morbidity and mortality, thus, requiring more intensive treatment than those with low severity scores. The purpose of the present study was to assess the severity of the acute abdomen in paediatric patients using a modification of the acute physiological and chronic ill-health evaluation II score (APACHE II). METHODS: Children admitted and operated on for generalized acute abdomen over a period of 7 years from January 1993 to December 1999 were prospectively studied. A study proforma was drafted and demographic, clinical, preoperative, operative and postoperative data on each patient were entered. Each patient had severity of illness assessed using APACHE II parameters with minor modification to make it applicable to children. Postoperative outcome and severity of illness were compared to determine any correlation. RESULTS: There were 69 patients operated on within the period of the study. Age ranged from 3 months to 15 years, with a mean of 9.1 SD 4.3 years. Forty-two patients (61.2%) were male and 27 (39.8%) were female. Typhoid intestinal perforation accounted for 35 (50.7%) and intestinal obstruction with or without intestinal gangrene accounted for nine (13%). Modified APACHE II score ranged from 0 to 18, mean 8.5 SD 5. For survivors, the mean score was 8; for non-survivors, 13. Eight patients died (11.6%): four of 63 (6.4%) patients who scored 0-15 died; four of six (66.7%) patients who scored 16-18 (P < 00.05) died. A modified APACHE II score greater than 15 was associated with a significantly greater mortality. The data for postoperative morbidity and hospital stay were not conclusive. CONCLUSION: Although the APACHE II score was designed for adults, a modification can be suitably applied to predict mortality in children with generalized peritonitis. There will be a need to apply this to large number of patients in order to validate our finding. PMID- 12752282 TI - Current attitudes to total hip replacement in younger patients: a comparison of two nations. AB - INTRODUCTION: Orthopaedic surgeons performing total hip replacements (THR) today are faced with a vast array of options. Inspired by a recent UK study, we wanted to determine the current trend in prosthesis choice, fixation and bearing surfaces used in 'young' Australian patients, and to compare this trend to the UK. METHODS: A questionnaire identical to that used in the UK study was posted to all current members of the Australian Orthopaedic Association and returned questionnaires were directly compared to the UK results on a percentage-of responses basis. RESULTS: Two hundred and forty-six valid responses were received. The number of THR reported to be performed by these respondents (15 789) was equivalent to the estimated number of prostheses sold here during the same period (15 624). The UK results showed a predominant use of Charnley and Exeter femoral prostheses, an all-polyethylene acetabular component, and cement fixation of both the acetabular and femoral components for both their older and younger patients. In younger patients, Australian surgeons favoured uncemented fixation techniques for the femur (57%vs 23%), and especially the acetabulum (85%vs 32%). There was a higher percentage use of modular design (95%vs 67%) and a very high use of ceramic as a bearing surface, 49% (vs 25%) using it for the femoral head, and 21% (vs 2%) employing a ceramic-on-ceramic bearing combination. DISCUSSION: Despite being privy to the same published papers, the THR prosthesis and fixation preferences of UK and Australian orthopaedic surgeons are markedly different. This may be because of interpretation of papers, peers, personal experience, patient assessment, budgets, institutions, theories, fashions, differences in autonomy and advertising. PMID- 12752283 TI - Embryonic parathyroid rests occur commonly and have implications in the management of secondary hyperparathyroidism. AB - BACKGROUND: Recurrence after surgery for secondary hyperparathyroidism is not infrequent. Regrowth of the residual parathyroid tissue after subtotal parathyroidectomy or of the autograft after total parathyroidectomy occurs in many cases. Supernumerary glands are also frequently cited as the offending cause and upon revisiting the neck, the surgeon may be surprised that such an obvious gland was 'missed' at the first operation. Indeed, multiple glands removed in sequential operations have been reported suggesting that they develop over time rather than being present from the start. It is possible that microscopic parathyroid 'rests' of embryological origin proliferate under the ongoing stimulus of renal failure to produce supernumerary glands after apparently adequate initial surgery for hyperparathyroidism. The aim of the present study was to determine whether such rests occur frequently or infrequently. METHODS: Operative details and pathology results from 60 consecutive parathyroidectomies were reviewed and the occurrence of parathyroid rests noted. RESULTS: Parathyroid rests were found in 37% of extra parathyroidal tissues submitted for analysis. These rests were found commonly in the thymus. The potential significance of such parathyroid rests undergoing hyperplasia in response to the ongoing stimulus of renal failure and leading to recurrent hyperparathyroidism is discussed. CONCLUSION: Parathyroid rests are common and potentially serve as a cause of recurrent disease in secondary hyperparathyroidism. Consideration should be given to performing thymectomy as part of the treatment of secondary hyperparathyroidism. A more detailed study is warranted. PMID- 12752284 TI - Men with colorectal cancer are predisposed to prostate cancer. AB - BACKGROUND: The objective of the present study was to investigate the relationship between colorectal and prostate cancer. METHODS: All Victorian men who developed metachronous colorectal and prostate cancer with the first primary diagnosed between 1982 and 1993 were identified retrospectively from the Victorian Cancer Registry and were followed up to the end of 1995. Analyses were stratified by age group and years of follow up. The cause of death in those men who had prostate cancer following colorectal cancer was determined. The stage of colorectal cancer was compared between men with and without second primary prostate cancer and the grade of prostate cancer was compared with men who did not have a prior colorectal cancer. RESULTS: Men who develop colorectal cancer are at increased risk of prostate cancer, with the greatest risk in men under the age of 65 (Relative risk approximately 2). Men with first primary colorectal cancer are more likely to develop prostate cancer than colorectal second primaries, and men who develop second primary prostate cancer are more likely to die of prostate cancer than colorectal cancer. CONCLUSIONS: Younger men diagnosed with colorectal cancer are at increased risk of prostate cancer. However, there is no direct evidence that screening for prostate cancer leads to a reduction in mortality. This should be considered when discussing long-term follow up. PMID- 12752285 TI - Radical lymph node dissection for melanoma. AB - BACKGROUND: Therapeutic lymph node dissection for melanoma aims to achieve regional disease control. Radical lymphadenectomy (RLND) can be a difficult procedure associated with significant postoperative morbidity. The aims of the present study were to review regional disease control and morbidity in a series of lymphadenectomies performed within a specialist unit. METHODS: The present study involved the analysis of 73 lymphadenectomies in 64 patients, from 1995 to 2001. RESULTS: The overall wound complication rate after inguinal lymphadenectomy (71%) was higher than after axillary lymphadenectomy (47%; P = 0.05). After inguinal lymphadenectomy, the wound infection rate was higher (25.0%vs 5.9%; P = 0.03), delayed wound healing was more frequent (25.0%vs 5.9%; P = 0.03), and the mean time that drain tubes remained in situ was longer (12.5 vs 8.2 days; P = 0.05). There were no significant differences in seroma (46%vs 32%) rates. Lymphoedema was more common after inguinal lymphadenectomy (P < 0.02). Multivariate analysis identified inguinal RLND (P = 0.002) and increasing tumour size (P = 0.045) as predictors of wound morbidity. More patients received postoperative radiotherapy after neck RLND compared to inguinal or axilla RLND (P = 0.03). Six (8%) patients developed local recurrence after lymphadenectomy. At a median follow up of 22 months, 34 (53%) patients have died, from disseminated disease. CONCLUSIONS: Radical lymphadenectomy for melanoma is associated with significant morbidity. Inguinal node dissection has a higher rate of complications than axillary dissection. Low local recurrence rates can be achieved, limiting the potential morbidity of uncontrolled regional metastatic disease. PMID- 12752286 TI - Clinical outcome following total laryngectomy for cancer. AB - BACKGROUND: Patients with advanced cancers of the larynx and hypopharynx have been treated with total laryngectomy at the Department of Head and Neck Surgery, Royal Prince Alfred Hospital, Sydney in the past. Increasingly, these patients are being managed with organ-sparing protocols using chemo-therapy and radiotherapy. The aim of the present study was to review complication, recurrence and survival rates following total laryngectomy. METHODS: Patients who had total laryngectomy for squamous carcinomas of the larynx or hypopharynx between 1987 and 1998 and whose clinicopathological data had been prospectively accessioned onto the computerized database of the Department of Head and Neck Surgery, Royal Prince Alfred Hospital, were reviewed. Patients whose laryngectomy was a salvage procedure for failed previous treatment were included. RESULTS: A total of 147 patients met the inclusion criteria for the study, including 128 men and 19 women with a median age of 63 years. Primary cancers involved the larynx in 90 patients and hypopharynx in 57. There were 30 patients who had recurrent (n = 24) or persistent disease (n = 6) after previous treatment with radiotherapy (26 larynx cases and four hypopharynx cases). Pharyngo-cutaneous fistulas occurred in 26 cases (17.7%) and, using multivariate analysis, the incidence did not correlate with T stage, previous treatment or concomitant neck dissection. Local control rates were 86% for the larynx and 77% for the hypo-pharynx groups and neck control was 84% and 75%, respectively. Five-year survival for the larynx cancer group was 67% and this was significantly influenced by T stage and clinical and pathological N stage. Survival in the hypopharynx group was 37% at 5 years and this did not significantly correlate with T or N stage. There was a non significant trend to improved survival among previously treated patients whose laryngectomy was a salvage procedure. CONCLUSION: Patients with cancer of the larynx had a significantly better survival following total laryngectomy than patients with hypopharyngeal cancer. Those whose laryngectomy was carried out as a salvage procedure following failed previous treatment did not have a worse outcome than previously untreated patients. PMID- 12752287 TI - Biliary access loops for intrahepatic stones: results of jejunoduodenal anastomosis. AB - BACKGROUND: Patients with intrahepatic calculi require multiple interventions following successful surgical stone clearance for recurrent stones and cholangitis. The present paper describes the results of a technique of in continuity side-to-side jejunoduodenal anastomosis (JDA) that provides endoscopic access to the hepaticojejunostomy and intrahepatic ducts. This operation is compared to other techniques in a critical appraisal of various biliary access procedures described for long-term management of intrahepatic -calculi. METHODS: A retrospective analysis of clinical data of 13 patients who underwent biliary drainage procedures with access loops for intrahepatic calculi during the period March 1990 to December 2000 was performed. The postoperative course of patients and the feasibility of postoperative endoscopic access to the hepaticojejunostomy and intrahepatic ductal system in treatment of recurrent cholangitis were assessed. Nine patients underwent JDA, two underwent permanent-access hepaticojejunostomy (PAH) and two others underwent an interposition hepaticojejunoduodenostomy (IHJ). RESULTS: The analysis revealed no major procedure-related complications or mortality. Endoscopic access (using forward viewing gastroscope) was possible in 100% of cases following JDA, and with difficulty in both cases after PAH. Endoscopic access in the two patients with IHJ failed because of technical reasons. Recurrent cholangitis was seen in seven patients (54%) - two out of two patients in the PAH group, one out of two in the IHJ group and four out of nine in the JDA group. This required 12 endotherapy sessions (mean: 1.5 procedures per patient). CONCLUSION: In-continuity side-to side JDA allows easy access of conventional gastroduodenoscopes to the biliary tree for removal of recurrent/residual intrahepatic stones. The technique has advantages over other access loop procedures in the long term management of recurrent intrahepatic stones. PMID- 12752288 TI - In vitro comparison of glyceryl trinitrate-verapamil with other dilators of human saphenous vein. AB - BACKGROUND: A common problem encountered during harvesting of the saphenous vein for coronary artery bypass grafting (CABG) is spasm of the vein. The previous work of the present authors has led to the development of a dilator solution containing glyceryl trinitrate (GTN) and verapamil for use during harvesting of the saphenous vein (SV) and the internal mammary artery. The present organ bath study was designed (i) to determine the best vasodilator agents for the saphenous vein during harvesting and (ii) to investigate the presence of any synergistic effect when vasodilator agents are combined. METHODS: Discarded segments of saphenous vein were collected from 46 patients undergoing CABG and studied in organ baths. Initially veins were contracted with potassium or thromboxane A2 mimetic (U46619). Dilators were added individually or in combination in clinically relevant concentrations to study time of onset and maximum effect. The vasodilator agents tested included GTN, verapamil (V), sodium nitroprusside (SNP) and nicorandil (N). RESULTS: When rapidity of onset (i.e. relaxation caused in the first minute) was considered, GTN or combinations containing GTN, caused a response that was significantly faster than the other vasodilators (P < 0.05). The maximum response caused by the GTN-V combination (61.4 +/- 9.6%) was significantly greater than all the other single dilators, SNP (35 +/- 4%) and N (10 +/- 3%) (P < 0.05). However, no synergistic effects were demonstrated. CONCLUSIONS: The present paper demonstrates the following conclusions: (i) glyceryl trinitrate is a rapidly acting dilator of the human isolated SV. (ii) GTN-V combination is a potent combination causing a maximum dilator effect that is significantly greater than single agents including SNP and N. PMID- 12752289 TI - Day-only laparoscopic cholecystectomy in a regional teaching hospital. AB - BACKGROUND: Day-only laparoscopic cholecystectomy (DOLC) has been demonstrated to be a safe and feasible procedure. The aim of the present study was to introduce DOLC to a busy teaching hospital without a separate day-surgery facility, to identify any problems associated with early discharge, and to determine patient satisfaction. METHODS: Over a 2-year period, all patients undergoing elective laparoscopic cholecystectomy under one surgeon were prospectively studied. Patients satisfying criteria for DOLC were offered the procedure. All patients were sent anonymous satisfaction surveys postoperatively. RESULTS: One hundred and one patients underwent elective laparoscopic cholecystectomies and 41 of these patients were booked for DOLC. Thirty-three (80%) were successfully discharged the same day and there were no complications related to early discharge. Only two of eight unplanned admissions were because of postoperative pain or nausea. Thirty-two (78%) of DOLC patients replied to our survey and of those, 24 (78%) were satisfied with their length of stay. The extra strain placed on day-stay ward resources was reflected in patient survey comments on their care. CONCLUSIONS: Our findings support the evidence that DOLC is safe and feasible. However, in a busy teaching hospital with tight budget constraints and no separate day-surgery facility we found many patients satisfied with their length of stay but not always with the quality of care they received on the day stay ward. The latter was insufficiently equipped to handle procedures of this complexity. So although in theory DOLC has many advantages, we are unable to institute this as routine practice at this time. PMID- 12752290 TI - Laser tenotomy and vocal process resection for bilateral midline vocal fold fixation. AB - BACKGROUND: The present retrospective study presents the outcomes of a group of 21 patients with laryngeal obstruction caused by bilateral vocal fold fixation. All of these patients were treated by laser assisted muscle tenotomy and vocal process resection - a modification of the technique described by Michael and Eugene Rontal in 1994.1 METHODS: Between January 1997 and March 2002 the senior author performed muscle tenotomy and vocal process resection for bilateral vocal fold fixation on 21 patients. RESULTS: The mean follow-up time was 2.3 years. The technique was successful in achieving an adequate airway and good voice with no aspiration in every case. CONCLUSIONS: Laser assisted muscle tenotomy and vocal process resection is a proven treatment for bilateral medial vocal fold fixation with the provision of a good airway, good voice and the avoidance of aspiration. PMID- 12752291 TI - Spinal and spinal cord injuries in horse riding: the New South Wales experience 1976-1996. AB - OBJECTIVES: The objective of the present study was to determine the incidence of acute spinal cord injuries (ASCI) in all forms of horse riding in New South Wales (NSW) for the period 1976-1996. Other aims of the present study were to compare and contrast ASCI with vertebral column injuries (VCI) without neurological damage and to define appropriate safety measures in relation to spinal injury in horse-riding. DESIGN: A retrospective review was done of all ASCI cases (n = 32) admitted to the two acute spinal cord injury units in NSW for the cited period. A comparable review of VCI cases (n = 30) admitted to these centres for the period 1987-1995 was also undertaken. RESULTS: A fall in flight was the commonest mode of injury in both groups. Occupational and leisure riding accounted for 88% of ASCI and VCI. The incidence of ASCI is very low in those riding under the aegis of the Equestrian Federation of Australia - two cases in 21 years; and there were no cases in the Pony Club Riders or in Riding for the Disabled. The difference in the spinal damage caused by ASCI and VCI is in degree rather than kind. Associated appendicular/visceral injuries were common. CONCLUSIONS: No measures were defined to improve spinal safety in any form of horse riding. The possible role of body protectors warrants formal evaluation. Continued safety education for all horse riders is strongly recommended. PMID- 12752292 TI - Pierre Dionis and the Franco-British dialogue in surgery. AB - Pierre Dionis (1643?-1718), though not himself a very innovative surgeon, has been acclaimed as the author of a surgical textbook that opened a new era in which French surgical teaching dominated Europe. This dominance is reflected in the Gordon Craig Library, which contains four copies of the book by Dionis in English translation, as well as a number of other books by influential French 18th century surgical writers, notably Henri Francois Le Dran and Francois Quesnay. It is significant that many of these writers benefited from government patronage in various forms. Dionis himself began his career as demonstrator in anatomy and surgery at the Jardin du Roi, in his time the premier teaching institution in those sciences. He later became a court surgeon. During the course of the 18th century, French surgery gained ascendancy over French academic medicine and also on the inter-national stage. English surgery, likewise, made great progress. There was a very productive dialogue between medical scientists and teachers in both countries, assisted by textbooks in translation. The French Revolution demolished the medical and surgical institutions established under the old regime, but French surgery emerged from the rubble to play a great part in the birth of modern clinical medicine PMID- 12752293 TI - Anthony Eden's bile duct: portrait of an ailing leader. PMID- 12752294 TI - A triple tie: the relationship between science, religion and society. PMID- 12752296 TI - Inguinal hernia repair: where to next? PMID- 12752297 TI - Simplified technique for laparoscopic appendicectomy. PMID- 12752298 TI - Simplified technique for laparoscopic appendicectomy. PMID- 12752299 TI - Probabilities relevant to detecting poorly performing -trainees. PMID- 12752300 TI - Developmental anomaly of the inferior vena cava. AB - Despite the many possible modes of presentation, congenital anomalies of the inferior vena cava are increasingly being found in asymptomatic patients. Although plain chest radiography may reveal enough suggestive signs, the diagnosis is usually confirmed by ultrasound, computed tomography and magnetic resonance imaging. The authors present a case of infrarenal absence of the inferior vena cava that presented as a retroperitoneal mass and discuss the embryology and clinical implications of this unusual entity. PMID- 12752301 TI - Catastrophic rectal bleeding from a ruptured splenic artery aneurysm. PMID- 12752302 TI - Testicular microlithiasis: a case report and review of the literature. AB - Testicular microlithiasis (TM) is a rare condition in which men have innumerable testicular calcifications. It is increasingly being reported on ultrasound. The published literature has reported an association between confirmed testicular malignancy and testicular microlithiasis. The relationship between TM and the risk of developing malignancy is unclear. The present paper reports a patient with a previously normal scrotal ultrasound except for bilateral sonographically detected TM who developed a testicular tumour. It also discusses the appropriate management of TM after reviewing the published literature. PMID- 12752304 TI - Alternative T-cell costimulatory pathways in transplant rejection and tolerance induction: hierarchy or redundancy? PMID- 12752305 TI - Diabetes mellitus after kidney transplantation. PMID- 12752306 TI - Guidelines on the firing-line. PMID- 12752307 TI - Regulatory T cells in the control of transplantation tolerance and autoimmunity. AB - A role for immunoregulatory T cells in the maintenance of self-tolerance and in transplantation tolerance has long been suggested, but the identification of such cells has not been achieved until recently. With the characterisation of spontaneously occurring CD4+CD25+ and NK1.1+ T subpopulations of T cells as regulatory cells in rodents and in humans, together with several in vitro generated regulatory T-cell populations, it seems possible that 'customised' regulatory cells possessing antidonor specificity may become therapeutic tools in clinical transplantation tolerance. PMID- 12752308 TI - T-cell allorecognition and transplant rejection: a summary and update. AB - Transplant biologists have made significant progress over the last 20 years towards unraveling the immunologic intricacies of allograft rejection. This large body of work has resulted in an improved understanding of T-cell allorecognition at a molecular level and has provided new insight into the functional consequences resulting from the allorecognition events. The findings suggest that the survival and the histologic features of a transplanted organ are influenced not only by the T-cell recognition pathway, but also by the frequency, the induced effector functions and the specific cellular targets of the alloreactive T-cell repertoire. PMID- 12752309 TI - Mycophenolic acid pharmacodynamics and pharmacokinetics provide a basis for rational monitoring strategies. PMID- 12752310 TI - Role of 4-1BB in allograft rejection mediated by CD8+ T cells. AB - Blockade of traditional costimulatory molecules fails to inhibit rejection in many models where CD8+ T cells are sufficient to mediate rejection. This observation demonstrates that in many settings CD8+ T cells are not dependent upon CD28 or CD154 signals to mediate rejection. 4-1BB (CD137) has been shown to be an important regulatory molecule for CD8+ T cells in a variety of nontransplant models. Here we show that blocking the 4-1BB pathway significantly inhibited rejection of intestinal allografts by CD8+ but not CD4+ T cells. This effect was associated with significantly decreased expression of the genes encoding TNFalpha and secondary lymphoid chemokine (SLC) within the spleens of recipient mice. Disruption of the 4-1BB pathway also impaired the priming of alloantigen-specific CD8+ T cells and the accumulation of recipient dendritic cells within the spleen. These data directly demonstrate an important role for 4 1BB in allograft rejection; particularly rejection mediated by CD8+ T cells. Our data suggest that in addition to providing a direct costimulatory signal to T cells, the 4-1BB pathway may regulate other important steps in the immune response such as the migration of T cells and dendritic cells. PMID- 12752311 TI - Feline immunodeficiency virus-mediated viral interleukin-10 gene transfer prolongs non-vascularized cardiac allograft survival. AB - Previous experiments demonstrated plasmid-, retroviral-, or adenoviral-mediated vIL-10 gene transfer could prolong allograft survival, but transgene expression was rapidly extinguished. Feline immunodeficiency virus (FIV) can integrate into genomic DNA of nondividing cells, resulting in indefinite transgene expression. We hypothesized FIV-mediated gene transfer could provide long-term gene expression, and improved allograft survival. FIV-vIL-10 and FIV-beta-gal were produced using the FELIX vector system. With vector transfer to syngeneic cardiac grafts, beta-galactosidase reporter gene expression was noted as early as day 5, was strongly expressed at days 10 and 20, and persisted for 50 days after transplantation. For allografts, FIV-vIL-10 gene transfer more than doubled mean survival from 10 +/- 1.6 to 22.3 +/- 3 days. When combined with other immunosuppressants, such as anti-CD40L mAb, FTY720, or anti-CD3 mAb, the mean survival times were prolonged to 27 +/- 4.6 days, 27.8 +/- 4.6 days, and 45.5 +/- 4.9 days, respectively. Multiple chemokine and chemokine receptor genes were induced by ischemia-reperfusion injury in syngeneic grafts, and in allogeneic grafts more genes were induced and to a greater degree. In allogeneic grafts transduced with FIV-IL-10, a number of the chemokine genes were suppressed. Therefore, FIV virus-mediated vIL-10 gene transfer prolongs allograft survival and, in combination with other agents, produces an additive effect. PMID- 12752312 TI - Protective effect of the immunosuppressant sirolimus against aortic atherosclerosis in apo E-deficient mice. AB - Atherosclerosis is a chronic inflammatory disease that develops in response to injury to the vessel wall, and is augmented by hypercholesterolemia. To further delineate the role of the immune system and local factors in this process, we assessed the effects of the immunosuppressant sirolimus (Rapamycin, RAPAMUNE, Wyeth, Collegeville, PA) on atherosclerosis in the apoE-deficient (apoE KO) mouse, a well-accepted model of cardiovascular disease. ApoE KO mice were fed a high fat diet and sirolimus was administered. After 12 weeks, atherosclerotic lesions and plasma lipoproteins were measured. The expression of cytokines associated with atherosclerosis was also examined. All groups demonstrated plasma total cholesterol (TC) >1100 mg/dL. Sirolimus treatment was associated with a 30% increase in LDL-cholesterol (LDLc) and a dose-dependent elevation in HDL cholesterol (HDLc). Despite increased LDLc, aortic atherosclerosis was markedly reduced in all sirolimus-treated groups. Sirolimus treatment resulted in decreased expression of IL-12p40, IFN-gamma and IL-10 mRNA. In contrast, TGF beta1 was elevated. Sirolimus significantly reduced atherosclerosis in apo E-KO mice; this effect is independent of, and obviates, elevated plasma TC and LDLc. Sirolimus might therefore be of benefit on atherosclerosis in patients undergoing therapy, independent of any impact on circulating lipids. PMID- 12752313 TI - Acute ischemia/reperfusion injury after isogeneic kidney transplantation is mitigated in a rat model of chronic renal failure. AB - The influence of chronic renal failure on renal susceptibility to an acute ischemic insult was evaluated. Recipient Lewis rats were randomly assigned to undergo 5/6 nephrectomy (chronic renal failure, CRF) or sham operation (normal renal function, NRF). After 11 weeks, normal kidneys of Lewis donor rats were transplanted in the recipients. The outcome of the isografts was assessed. Filtration capacity of the isografts in the CRF rats was preserved to approximately one-quarter of its normal capacity on the 1st day post transplantation, whereas it fell to 0 in the NRF rats. This was reflected by a significantly higher increase in serum creatinine in the latter group. The isografts in the CRF rats had a significantly lower degree of acute tubular necrosis and no increase in the number of macrophages and T lymphocytes in the first 24 h in contrast to the NRF rats. Epithelial regeneration and repair started earlier in the CRF group. In conclusion, the present study indicated that CRF blunted ischemia/reperfusion injury of a transplanted kidney, and that its regeneration capacity was certainly not hampered by the presence of chronic uremia. These results will be the basis for studies on modulation of early leukocyte-endothelial interactions resulting from immunological disturbances inherent to the uremic environment. PMID- 12752314 TI - Prevention of chronic rejection with immunoregulatory cells induced by intrathymic immune modulation with class I allopeptides. AB - Intrathymic immune modulation with RT1.Aa allopeptides in the PVG.R8-to-PVG.1 U rat strain combination leads to long-term survival of cardiac allografts. This regimen, however, does not induce transplantation tolerance, since most long surviving allografts undergo chronic rejection. We investigated recipients with chronic rejection for donor-specific immune nonresponsiveness and immunoregulatory cells as possible mechanisms responsible for long-term graft survival. There was a significant reduction in the proliferative response of T cells from long-term allograft recipients to donor alloantigens as compared with that of naive T cells. Adoptive transfer of splenocytes from intrathymically manipulated primary long-term graft survivors into minimally irradiated secondary hosts resulted in indefinite survival of > 80% of allografts, providing evidence for immunoregulatory cells. Secondary recipients had total absence of donor reactive cellular and humoral responses. Immunoregulation was also transferable from secondary to tertiary graft recipients. More importantly, there was a significant reduction in the incidence of chronic rejection in secondary hosts (> 85%) and complete prevention of acute and chronic rejection in tertiary hosts. This study demonstrates that intrathymic immunomodulation with class I allopeptides results in the generation of immunoregulatory cells that do not block chronic rejection in primary hosts where they develop, but prevent both acute and chronic allograft rejection when adoptively transferred into secondary and tertiary recipients. PMID- 12752315 TI - Incidence and cost of new onset diabetes mellitus among U.S. wait-listed and transplanted renal allograft recipients. AB - This study sought to determine 1) the incidence and costs of new onset diabetes mellitus (NODM) associated with maintenance immunosuppression regimens following renal transplantation and 2) whether the mode of dialysis pretransplant or the type of calcineurin inhibition used for maintenance immunosuppression affected either the incidence or cost of NODM. The study examined the United States Renal Data System's clinical and financial records from 1994 to 1998 of all adult, first, single-organ, renal transplantations in either 1996 or 1997 with adequate financial records. It used the second diagnosis of diabetes in previously nondiabetic patients to identify NODM. While NODM had an incidence of approximately 6% per year among wait-listed dialysis patients, NODM over the first 2 years post-transplant had an incidence of almost 18% and 30% among patients receiving cyclosporine and tacrolimus, respectively. By 2 years post transplant, Medicare paid an extra $21 500 per newly diabetic patient. We estimated the cost of diabetes attributable to maintenance immunosuppression regimens to be $2025 and $3308 for each tacrolimus patient and $1137 and $1611 for each cyclosporine patient at 1 and 2 years post-transplant, respectively. PMID- 12752316 TI - Histological grading of chronic pancreas allograft rejection/graft sclerosis. AB - Chronic rejection (CR) of pancreas allografts needs to be accurately defined and diagnosed. We propose a grading scheme designed for percutaneous needle biopsies (C0-CIII). Grading is based on the semi-quantitative determination of fibrosis and corresponding proportional loss of exocrine parenchyma. Pancreas biopsies (n = 141) from 46 patients were studied. Twenty-six patients lost graft function after a mean time of 25.5 months, whereas 20 patients retained good graft function during a mean follow-up of 67.7 months. Sequential biopsies showed gradual progression of CR over time (p = 0.0001), and good correlation was found between the CR grade and the time elapsed from transplantation (p = 0.0001). The CR grade was predictive of the remaining time of graft function (54.3 months for C0, 24.6 months for CI, 9.7 months for CII and 1.6 months for CIII p = 0.00001). Preceding episodes of acute rejection (AR) were more frequent and more severe, and often occurred late in patients with graft loss due to CR (p = 0.04). Reproducibility among pathologists from different institutions was excellent for grades C0 (Kappa 0.9) and CIII (0.87), substantial for Grade CII (0.61) and moderate for Grade CI (0.59). The proposed grading scheme provides a reliable and reproducible tool for the assessment of CR in percutaneous needle biopsies, with definite prognostic significance. PMID- 12752317 TI - Pharmacokinetics of an everolimus-cyclosporine immunosuppressive regimen over the first 6 months after kidney transplantation. AB - The pharmacokinetics of everolimus were characterized over the first 6 months post transplant in 731 patients receiving either 0.75 or 1.5 mg bid everolimus in addition to cyclosporine and corticosteroids. Pharmacokinetic data consisted of 4014 everolimus trough concentrations (Cmin) obtained in all patients and 659 area under the concentration-time curve (AUC) -profiles obtained at months 2, 3, and 6 in a subset of 261 patients. Cmins averaged 4.3 +/- 2.4 and 7.2 +/- 4.2 ng/mL at 0.75 and 1.5 mg bid, indicating a 20% under-proportionality at the upper dose level. Cmins were 19-34% lower in the first month compared with months 2 through 6-values. AUC was dose-proportional and stable over time, averaging 77 +/ 32 and 136 +/- 57 ng.h.mL-1 at the two dose levels. Within- and between-patient variability in AUC were 27% and 31%, respectively. There was no influence of sex, age (16-66 years), or weight (42-132 kg) on AUC. Everolimus exposure was significantly lower by an average 20% in blacks. Everolimus exposure was relatively stable over the first 6 months post transplant, with no major departure from dose-proportionality over the therapeutic dose range. Weight adjusted dosing (mg/kg) does not appear warranted. Black patients may have lower bioavailability and/or higher clearance of everolimus compared with white patients. PMID- 12752318 TI - Non-heart beating donor kidneys with delayed graft function have superior graft survival compared with conventional heart-beating donor kidneys that develop delayed graft function. AB - Delayed graft function may have an association with reduced graft survival, and nonheart-beating donor (NHBD) kidneys have higher rates of delayed graft function (DGF) than heart-beating donor (HBD) kidneys. This study compared outcome of renal transplants from HBDs who developed DGF, with NHBDs who developed DGF. All recipients of HBD and NHBD kidneys who developed DGF were identified during a 10 year period. All patients with graft primary nonfunction were excluded from analysis. Four hundred and fifty-six functioning transplants were performed. Delayed graft function occurred in 69 (17%) HBD and 55 (93%) NHBD kidneys. The grafts developing DGF were well matched for donor and recipient age. The rate of acute rejection was similar; [n = 16/69 (23%) HBD vs. n = 13/55 (24%) NHBD]. Cold ischaemia was 21 h in the HBD group and 17 h in NHBD group (p > 0.05). Serum creatinine was similar for both groups at 1.3 and 6 years (p > 0.05 for all time points). Graft survival in the NHBD recipients with DGF was significantly better at 3 years (84%) compared with recipients of a HBD renal transplant that developed DGF (73%) (p < 0.05), and at 6 years (62% survival for HBDs and 84% survival for NHBDs). This study shows that graft survival was better for NHBD kidneys up to 6 years after transplantation. PMID- 12752319 TI - Screening for prostate, breast and colorectal cancer in renal transplant recipients. AB - American Society of Transplantation guidelines recommend screening renal transplant recipients for breast, colorectal and prostate cancer. However there is a lack of evidence to support this practice. Computer simulation modeling was used to estimate the years of life lost as a result of these cancers in 50-year old renal transplant recipients and subjects in the general population. Renal transplant recipients lost fewer years of life to cancer than people in the general population largely because of reduced life expectancy. In nondiabetic transplant recipients, loss of life as a result of these cancers was comparable with that in the general population only under assumptions of increased cancer incidence and cancer-specific mortality risks. Even with two-fold higher cancer incidence and disease-specific mortality risks, diabetic transplant recipients lost considerably fewer life years to cancer than those in the general population. Recommended cancer screening for the general population may not yield the expected benefits in the average renal transplant recipient but the benefits will be considerably higher than for patients on dialysis. Transplanted patients at above-average cancer risk in good health may achieve the benefits of screening that are seen in the general population. PMID- 12752320 TI - Pretransplant MELD score as a predictor of outcome after liver transplantation for chronic hepatitis C. AB - The Model of End-Stage Liver Disease (MELD) score, an accurate predictor of mortality in patients awaiting liver transplantation (OLTX), did not predict graft or patient survival in the post-transplant setting. Our aim was to test the model in patients who underwent OLTX for chronic hepatitis C. Two hundred and eighty-seven adult patients who underwent primary OLTX for chronic hepatitis C between December 1993 and September 1999 were studied from a prospectively maintained database. The group was stratified by MELD scores of less than 15, 15 24, and greater than 24. Patient survival, graft survival, and interval liver biopsy pathology were reviewed. Both patient and graft survival at 3, 6, and 12 months were significantly lower in the higher MELD score groups, as was patient survival at 24 months (p-values, 0.01-0.05). The difference in survival between the low, medium, and high MELD score groups increases in time. The survival without bridging fibrosis in the allograft at 1 year post-transplant was significantly lower with higher MELD scores (p = 0.037). The decrease in survival seen in hepatitis C patients with MELD scores greater than 24 raises questions of transplant suitability for these patients. Therapeutic modalities to decrease post-transplant graft injury in these patients should be explored. PMID- 12752321 TI - Kidney-pancreas transplantation in a long-term non-progressor HIV-infected recipient. AB - With the introduction of highly active antiretroviral therapy (HAART), HIV infection has become a chronic disease with more frequent end-stage organ failures. As a result, the question of transplantation in HIV patients is raised more often. Although still subject to controversies, HIV infection is no longer an absolute contraindication to solid organ transplantation. We report a case of combined kidney-pancreas transplantation in a HIV recipient. HIV has remained stable without any antiviral therapy for up to 2 years after transplantation and has reached criteria for inclusion in the long-term nonprogressor (LTNP) group. Grafted organs demonstrated good function without rejection. This case emphasizes the need to consider LTNP HIV patients as a specific subgroup, when discussing solid organ transplantation. HAART is not required, thus sparing drug interactions and their unique immunological features, such as CCR5 mutation, might prevent rejection. This subgroup of HIV patients should be offered less restricted access to transplantation. PMID- 12752322 TI - De-novo cholangiocarcinoma in the setting of recurrent primary sclerosing cholangitis following liver transplant. AB - Orthotopic liver transplantation is the only definitive therapeutic option in patients with primary sclerosing cholangitis (PSC) and end-stage liver disease. However, PSC recurs in up to 20% of patients transplanted for this indication. To date, no patient has been reported to develop cholangiocarcinoma (CCA) post transplant, without biliary tract cancer having been present pretransplant. Here, we report recurrent PSC complicated by de-novo CCA in a 31-year-old man transplanted for PSC 8 years earlier. Cholangiocarcinoma was confirmed using a combination of computed tomography, cholangiography, positron emission tomography and histological examination of biliary cytology. He has since been successfully re-transplanted following preoperative chemo-radiotherapy. No viable tumor was identified in the explanted liver. This case establishes that long-term complications associated with PSC and biliary-enteric surgery such as CCA may become apparent in new grafts post-transplant. PMID- 12752323 TI - Enhanced soluble interleukin-5 receptor alpha expression in nasal polyposis. AB - BACKGROUND: Alternative splicing of the interleukin-5 receptor alpha (IL-5Ralpha) subunit leads to the generation of a signalling, membrane-anchored (TM) isoform, or a secreted [soluble (SOL)], antagonistic variant. Given the key role of IL-5 in eosinophil function, we investigated SOL IL-5Ralpha expression pattern in an eosinophil-associated disease such as nasal polyposis (NP). METHODS: An SOL IL 5Ralpha enzyme-linked immunosorbent assay and quantitative real-time polymerase chain reaction (PCR) were established and applied in serum, nasal secretion and nasal tissue of controls (n = 12), and NP patients (n = 42) with or without asthma. RESULTS: Analysis of serum, nasal secretion, and nasal tissue samples revealed that SOL IL-5Ralpha protein concentrations were significantly increased in NP vs control tissue. Within the NP group, there was a significant up regulation of SOL IL-5Ralpha in patients with systemic airway disease. These findings were confirmed at the mRNA level, using an optimized real-time reverse transcriptase PCR procedure. CONCLUSIONS: This report demonstrates SOL IL-5Ralpha transcript and protein up-regulation in NP. Soluble IL-5Ralpha differentiates nasal polyps with or without concomitant asthma. As SOL IL-5Ralpha is strongly up regulated for disease and has antagonistic properties in vitro, our studies shed new light on the mechanisms of specific immunomodulatory therapies, such as anti IL-5. PMID- 12752324 TI - Allergen-reactive antibodies are found in nasal fluids from patients with birch pollen-induced intermittent allergic rhinitis, but not in healthy controls. AB - BACKGROUND: Increased levels of allergen-reactive immunoglobulins (Igs) have been reported in nasal fluids from patients with intermittent allergic rhinitis (IAR) sensitive to ragweed and grass. The aims of this study were to make a detailed characterization of nasal fluid Igs in birch pollen-induced IAR. METHODS: Nasal fluids were obtained from 23 patients with birch pollen-induced IAR during and after the birch pollen season, and from 20 healthy controls. Nasal fluid total and Bet v 1-reactive (IgA), IgE and IgG as well as albumin were analyzed by immunoassays. The integrity of IgA and IgG, and the molecular form of IgA were assessed by Western blotting and column fractionation, respectively. RESULTS: Nasal fluid total IgE and IgG, but not IgA, were higher in patients compared with controls. Western blotting indicated no significant degradation of IgA (including S-IgA) and IgG. Most of the IgA, including Bet v 1-reactive antibodies, was of the secretory form and of the IgA1 subclass. Bet v 1-reactive IgA and IgG were present in all patients, but was mostly nondetectable in controls. No significant differences in the levels of Bet v 1-reactive IgA and IgG were found in patients during the birch pollen season compared with off season. Both Bet v 1 and Bet v 2 reactive IgE were nondetectable in most samples. CONCLUSIONS: Nasal fluid Bet v 1 reactive IgA and IgG were found in all patients with birch pollen-induced IAR, but not in controls. However, no significant differences were found between patients during and after the birch pollen season. PMID- 12752325 TI - The IL1A genotype is associated with nasal polyposis in asthmatic adults. AB - BACKGROUND: Nasal polyposis (NP) is a chronic inflammatory disease often found coexisting with asthma. As this disorder tends to cluster in families, a genetic predisposition has been suggested. Interleukin-1 (IL-1) has been proposed to play a role in the pathogenesis of NP. METHODS: We analysed the single G-to-T base exchange polymorphism in exon 5 at +4845 of the gene encoding IL-1alpha (IL1A) and the C-to-T base exchange polymorphism at -511 of the gene encoding IL-1beta (IL1B) in a population-based sample of adult asthma patients (n = 245). The data were assessed for correlation with data on history of NP and other phenotype related characteristics. RESULTS: The prevalence of NP in our study group was 14.3%. The distribution of the IL1A genotype differed significantly between asthmatics with and without NP (P = 0.005). The risk of NP was markedly increased in allele G homozygous subjects (OR = 2.73; 95%CI = 1.40-5.32). In the case of IL1B we found no significant associations. Asthmatics with NP had more symptoms than others, but lung function and blood eosinophil counts were similar. CONCLUSIONS: Our study demonstrates an association of IL1A with NP inasthmatic patients and addresses the role of IL-1alpha as an inflammatory modulator in the pathogenesis of this disease. PMID- 12752326 TI - Inhibitory effects of ketotifen on eotaxin-dependent activation of eosinophils: consequences for allergic eye diseases. AB - BACKGROUND: The aim of this study was to investigate the effects of ketotifen on different parameters of human eosinophil functions, namely chemotaxis, oxidative metabolism and mediator release, induced after activation. METHODS: Eosinophils from hypereosinophilic patients or normal donors were purified by Percoll gradient and the magnetic cell separation system. Chemotaxis was studied using the Boyden chamber technique using three potent chemoattractants: formyl methionine-leucine-phenylalanine (fMLP), interleukin (IL)-5 and eotaxin. Oxidative metabolism was determined by a luminol-dependent chemiluminescence assay after activation with eotaxin or secretory immunoglobulin A (sIgA). The release of eosinophil cationic protein (ECP) and eosinophil derived neurotoxin (EDN) was measured by radioimmunoassay after activation with sIgA. RESULTS: At pharmacologically active concentrations and in a dose-dependent manner, ketotifen significantly inhibited the chemotaxis of eosinophils to fMLP, IL-5 and eotaxin. The production of reactive oxygen species induced by eotaxin and sIgA was decreased by ketotifen, showing a more pronounced effect when cells were activated by eotaxin. Activation by sIgA resulted in ECP and EDN release, which was partially inhibited by ketotifen. CONCLUSIONS: Through inhibition of chemotaxis, ketotifen might limit the number of eosinophils at the inflammation site during allergic reaction. Furthermore, inhibition by ketotifen of main inflammatory mediators release suggests a potential role of the drug in limiting the pathological potential of eosinophils. PMID- 12752327 TI - Polygalacturonase (pectinase), a new oilseed rape allergen. AB - BACKGROUND: Type I hypersensitivity to rapeseed pollen allergens was described as the result of a cross-sensitization with various pollens that could constitute an aggravating factor in birch or grass pollen allergies. Recently, a few rapeseed pollen allergens were described. The aim of the present work was to identify new rapeseed pollen allergens by using two-dimensional gel analysis, microsequencing, and mass spectrometry. METHODS: Water extractable proteins from oilseed rape pollen or stamen were separated by two-dimensional gel electrophoresis. The proteins were then electroblotted onto a nitrocellulose (NC) sheet. The NC sheets were successively incubated with (1) individual human sera pre-selected for their immunoglobulin E (IgE) reactivity to rapeseed pollen proteins, (2) alkaline phosphatase (AP)-conjugated goat anti-human IgE and (3) AP substrate. The allergens localized by this method were then identified by microsequencing and MALDI-TOF mass spectrometry analysis. RESULTS: Of the 18 sera studied, five recognized a wide multispot zone with a molecular mass around 43 kD and pIs between 6.5 and 8.5. The results obtained with two representative sera are shown. From this zone, two isoforms of the polygalacturonase enzyme were identified by microsequencing. Confirmation was obtained through MALDI-TOF mass spectrometry analysis. CONCLUSION: The present results allow the identification of a new rapeseed allergen that can be the main allergen for some patients. PMID- 12752328 TI - Comparative allergenicity studies of native and recombinant Blomia tropicalis Paramyosin (Blo t 11). AB - BACKGROUND: The complementary DNA (cDNA) encoding for Blo t 11, a 102 kD allergen from Blomia tropicalis (Bt) was isolated, expressed and characterized previously. This study aimed to isolate the native Blo t 11 allergen and compare its allergenicity with the recombinant forms. METHODS: Native Blo t 11 (nBlo t 11) was isolated from crude Bt extract by immuno-affinity chromatography, analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot, and verified by MALDI-TOF MS. Recombinant full-length Blo t 11 (rFL-Blo t 11) and its immunodominant peptide (fD) were expressed as glutathione S transferase (GST)-fusion proteins in Escherichia coli. Immunoglobulin E (IgE) reactivity of the Blo t 11 allergens were determined by enzyme-linked immunosorbent assay (ELISA) and skin prick test. The inhibition capacity of the nBlo t 11 against fD and vice versa was determined by absorption studies. RESULTS: Affinity purified nBlo t 11 was susceptible to degradation with the major degraded product resolved at approximately 66 kD. The nBlo t 11 was confirmed by immunoblot analysis and MALDI-TOF MS that generated 13 peptides with complete identity to the deduced amino acid sequence of Blo t 11. Comparative in vitro and in vivo allergenicity tests and the cross inhibition studies between the native and recombinant Blo t 11 showed that recombinant fD, but not the rFL Blo t 11, has comparable IgE reactivity with the native counterpart. CONCLUSIONS: This comparative study confirmed that the recombinant peptide fD contains the main immunodominant region of Blo t 11. This recombinant peptide, instead of the full-length protein, is a good candidate for diagnostic and therapeutics development for mite allergy. PMID- 12752329 TI - The epidemiology of atopic dermatitis in Italian schoolchildren. AB - BACKGROUND: Atopic dermatitis (AD) is common in children in industrialized countries. Only one large population study on its prevalence has been conducted in Italy, based on self-report questionnaire. The present study was designed to estimate the prevalence of AD in schoolchildren in Italy by dermatologists' assessment and by UK Working Party criteria, and to investigate associated symptoms and factors. METHODS: Cross-sectional survey on a random sample of 9 year-old schoolchildren from seven Italian cities. Children were examined by experienced dermatologists. Parents and teachers answered standardized questionnaires. RESULTS: Of the 1369 children examined, 88 had a diagnosis of AD, with an estimated point prevalence of 5.8% (95% CI 4.5-7.1) in the reference population. The reported lifetime prevalence was 15.2 (95% CI 12.2-18.2) for AD, 11.9% (95% CI 9.0-14.8) for asthma, and 17.6% (95% CI 14.6-20.7) for rhino conjunctivitis. The strongest associated factor was the presence of AD in at least one parent. No association of AD with maternal smoking during pregnancy, birth weight, maternal age at the time of the child birth and breast-feeding was observed. The environmental characteristics of the house and the school did not correlate with the prevalence of AD. Episodes of lower respiratory tract infections were associated with asthma, and to a lower extent also with AD and rhinitis. CONCLUSIONS: The prevalence of doctor-diagnosed AD in Italian schoolchildren is comparable to those reported for other developed countries. Family history of atopy was the single most important associated factor, while the complex interplay of environmental factors remains to be elucidated. PMID- 12752330 TI - Frequency and intensity of responses to mite patch tests are lower in nonatopic subjects with respect to patients with atopic dermatitis. AB - BACKGROUND: So far the issue of patch tests with mite allergens in subjects not affected by atopic dermatitis (AD) has been poorly investigated. The aim of this study was to assess the frequency and intensity of responses to atopy patch tests with Dermatophagoides in non-AD subjects, and to compare them to the ones observed in AD patients. METHODS: Patch tests were performed, employing a mixture of Dermatophagoides pteronyssinus and farinae at two different concentrations (20 and 40% pet.), on 75 non-AD subjects, including 33 patients with allergic rhinitis and 42 healthy volunteers, and on 210 AD patients, as controls. RESULTS: Positive responses to 20% mite patch tests were observed in 17 non-AD subjects (23%) and in 102 AD ones (49%). The former showed a lower intensity of the responses, with a mean score of 1.4 compared with 2.1 in atopics. CONCLUSIONS: Although positive responses to mite patch tests are also observable in subjects without AD, their frequency and intensity are significantly lower compared with AD patients. PMID- 12752331 TI - The effect of specific immunotherapy on T-cell receptor repertoire in patients with allergy to house-dust mite. AB - BACKGROUND: The precise mechanism of specific immunotherapy (SIT), long used for treating allergic diseases, remains undefined. SIT was shown to act by modifying the immune response of T lymphocytes to antigens. We examined the effect of SIT on the expression and use V-alpha, -beta, -gamma and -delta chains of T-cell receptors (TCR) in patients allergic to house-dust mite. METHODS: Peripheral venous blood was taken for lymphocyte TCR analysis from 10 house-dust mite (HDM) allergic adults before initiating SIT and 6 months after initiating the treatment. Twelve similarly allergic patients without SIT served as controls. TCR chains were identified by fluorescence-activated cell sorter (FACS) using the following monoclonal antibodies: CD3, CD14, CD8, pan alpha-beta, pan gamma-delta, V-alpha2, V-alpha12.1, V-beta5a, V-beta5b, V-beta5c, V-beta8a, V-beta8b, V beta3.1, V-beta13, V-beta12, V-beta6.7, V-delta1, V-delta2, V-gamma9, and V gamma4. RESULTS: Analyzed before and 6 months after SIT initiation, lymphocyte TCR showed significantly increased V-beta5b, V-beta12 and V-alpha12.1 values compared to controls (without significant changes in other markers). CONCLUSIONS: SIT caused selective expansion of certain V-beta- and V-alpha-expressing T cells in patients allergic to HDM. Our results support the notion that the effect of SIT in patients with allergic rhinitis may be achieved by modifying the T lymphocyte response through the modulation of TCR usage. PMID- 12752332 TI - How long does the effect of birch pollen injection SIT on apple allergy last? AB - BACKGROUND: Recent studies showed that injection specific immunotherapy (SIT) with birch pollen extract greatly reduces or cures the associated apple allergy in a large proportion of birch pollen-allergic patients. However, the long-term efficacy of SIT for apple allergy has not been assessed. OBJECTIVE: To evaluate the duration of the effect of injection SIT with birch pollen extract on apple allergy in birch pollen-allergic patients. METHODS: Thirty birch pollen-allergic patients showing both the clinical disappearance of apple allergy and a negative SPT with fresh apple at the end of their injection SIT course were followed-up at 12-month intervals from 6 months after SIT was stopped. Apple tolerance as well as SPT was assessed on all occasions. Fifty-seven birch pollen-allergic subjects without apple allergy and not submitted to SIT regularly followed-up for the onset of oral allergy syndrome (OAS) were used as controls. RESULTS: The overall prevalence of OAS after 30 months of follow-up did not differ between patients and controls. Although most patients became re-sensitized to apple by SPT over time, >50% of them were still able to tolerate eating the fruit at the 30-month follow-up visit. CONCLUSION: Although most patients show a 'natural', gradual propensity to apple re-sensitization (a consequence of prolonged and repeated inhalation of birch pollen responsible for primary sensitization?), the clinical effects of injection SIT on food allergy seem rather long lasting. PMID- 12752333 TI - Distinct delayed T-cell response to beta-methasone and penicillin-G in the same patient. AB - BACKGROUND: Multiple drug allergy syndrome is a clinical condition characterized by reactions against more than one different class of, both pharmacologically and structurally, unrelated drugs. Scanty data are available to date about a multiple drug delayed hypersensitivity syndrome. Our aim was to report the case of a delayed reaction to both beta-methasone (beta-MT) and penicillin-G (pen-G) occurring in the same patient, and analyse beta-MT- and pen-G-specific T-cell Lines (TCLs) with regard to their specificity, phenotype and cytokine profile. METHODS: We generated two drug-specific TCLs from biopsies at the site of positive intradermal reactions, and analysed their immunophenotype, T-cell receptor Vbeta (TCR-Vbeta) domains expression and cytokine profile. RESULTS: We demonstrated the specificity of the T cells isolated from positive intradermal test reactions to pen-G and beta-MT through the strict dose-dependent proliferation in response to drug-pulsed autologous antigen presenting cells. Fluorescence activated cell sorter (FACS) analysis revealed a predominance of CD4+ cells in the inflammatory cell infiltrate of intradermal test with beta-MT, while a predominance of CD8+ T cells in the site of delayed reaction to pen-G was found. The drug specific CD4+ and CD8+ T cells were heterogeneous, with regard to TCR-Vbeta usage. CD8+ pen-G-TCL displayed a preferential T helper 2 (Th2) profile, while a substantially heterogeneous pattern of cytokine production characterized specific beta-MT TCL. CONCLUSION: The study describes the coexistence in the same patient of a delayed hypersensitivity to both penicillin G and beta-MT, driven, respectively, by pen-G-specificTh2-skewed CD8+ and beta-MT specificTh0 CD4+ T cells. This case further support the existence of a multiple drug allergy syndrome also for delayed hypersensitivity. PMID- 12752334 TI - Severe drug rashes in three siblings simultaneously. AB - BACKGROUND: Adverse reactions to drugs commonly occur sporadically. Certain individuals seem to have an increased susceptibility to develop reactions to multiple drugs. Genetic predisposition has not been elucidated. Our objective was to describe a case of three siblings who developed severe rashes to the same drug simultaneously. METHODS: Review of the patients' medical records for information on the clinical course, and comparing their serum immunoglobulin (Ig) levels initially and during follow-up. RESULTS: The rashes were compatible with erythema multiforme and Stevens-Johnson syndrome and developed in the three siblings within 1-3 weeks after the intake of thiabendazole. Follow-up of serum Ig levels did not show any particular pattern, except for an initial mild to moderate elevation in IgG, IgA and IgM. CONCLUSIONS: The occurrence of such severe cutaneous drug reaction in the three siblings to the same drug suggests a genetic predisposition to adverse drug reactions. PMID- 12752335 TI - Specific sensitization to the common housefly (Musca domestica) not related to insect panallergy. AB - BACKGROUND: Allergy to houseflies is rare. We report a case of respiratory allergy from occupational exposure to houseflies in a farmer. CASE REPORT: A 30 year-old female farmer with a long-standing history of grass pollen allergy observed for 2 years rhino-conjunctivitis and mild asthma when entering livestock stables and barns. Allergy retesting revealed sensitization to various pollens but not to animal danders. Houseflies (Musca domestica) occurring on the farm in great quantity were suspected by the farmer herself as the causative agent. RESULTS: Skin prick testing with housefly was positive in the patient and negative in four controls. Experimental radioallergosorbant test was class 3 positive. Sensitization to house dust mite, storage mites and cockroach was not detectable. Western blots with housefly extracts revealed immunoglobulin E (IgE) binding to bands of 70, 50, and approximately 16 kDa. Tropomyosin in the housefly extract (35 kDa) was recognized by a tropomyosin reference serum but not by the patient. In enzyme-linked immunosorbent assay (ELISA) inhibition assays using housefly as the solid phase, IgE-binding of the patient was inhibited by 75% by M. domestica and by 44% by the closely related lesser housefly (Fannia canicularis), but not by extracts from blowfly (Lucilia spp.), fruit fly (Drosophila spp.), horsefly (Haematopota pluvialis) and mosquito (Culex pipiens). The IgE-binding of the tropomyosin control serum was inhibited by 60-80% by all species. CONCLUSIONS: In accordance with previous reports, this case demonstrates that respiratory sensitization to insects may be highly specific. According to ELISA inhibition, cross-sensitization in the present case was restricted to species of the family of true flies (Muscidae). PMID- 12752336 TI - Penicillin allergy. PMID- 12752337 TI - Laryngeal edema due to European bee-eater (Merops apiaster) in a patient allergic to honeybee. PMID- 12752338 TI - Antiphospholipid syndrome during allergic bronchopulmonary aspergillosis. PMID- 12752339 TI - Angioedema induced by angiotensin II blocker telmisartan. PMID- 12752340 TI - Adverse reactions linked to Quorn-brand foods. PMID- 12752341 TI - A case of isolated rice allergy. PMID- 12752342 TI - Type I sensitization towards patent blue as a cause of anaphylaxis. PMID- 12752343 TI - Successful fluorescein desensitization. PMID- 12752344 TI - Beta glucuronidase short-term immunotherapy. PMID- 12752345 TI - Review article: The advent of capsule endoscopy--a not-so-futuristic approach to obscure gastrointestinal bleeding. AB - Capsule endoscopy is a new, wireless, endoscopic examination of the small intestine. To date, two small clinical trials have been reported utilizing capsule endoscopy in patients with obscure gastrointestinal bleeding, and have shown its superiority to push enteroscopy in diagnosing the cause of blood loss. No outcome studies have been reported. This paper proposes a change in practice guidelines for obscure bleeding. It is our opinion that, in the future, with the advent of wireless capsule endoscopy, the evaluation of patients with obscure gastrointestinal bleeding will be very different from the practice of medicine today. We believe that capsule endoscopy will become the first-line method for the evaluation of patients with obscure bleeding, once upper endoscopy and colonoscopy have been shown to be negative. In patients with active bleeding, capsule endoscopy will confirm the small bowel as the site of bleeding, providing a location, or, if the study is negative for the small intestine, may indicate that the bleeding is either colonic or gastric in origin. In a patient with active bleeding within the small intestine, the capsule will guide further evaluation and therapy. A patient with a small bowel tumour detected by capsule endoscopy will proceed directly to laparoscopic surgery. If the site of bleeding is identified in the proximal small bowel and there is no mass, push enteroscopy will be used to re-identify the site and cauterize it. A distal small bowel site will require surgical intervention, coupled with intra-operative enteroscopy. Should the patient be too sick to undergo surgery, medical therapy utilizing hormonal agents will be considered. A colonic site will be evaluated by colonoscopy. In patients with a more occult or intermittent type of bleeding and in those whose upper endoscopies and colonoscopies are negative, capsule endoscopy will be used similarly to identify a bleeding lesion and thereby direct subsequent testing or treatment. PMID- 12752346 TI - Famotidine for infant gastro-oesophageal reflux: a multi-centre, randomized, placebo-controlled, withdrawal trial. AB - BACKGROUND: Gastro-oesophageal reflux afflicts up to 7% of all infants. Histamine 2 receptor antagonists are the most commonly prescribed medications for this disorder, but few controlled studies support this practice. AIM: To evaluate the safety and efficacy of famotidine for infant gastro-oesophageal reflux disease. METHODS: Thirty-five infants, 1.3-10.5 months of age, entered an 8-week, multi centre, randomized, placebo-controlled, two-phase trial: first 4 weeks, observer blind comparison of famotidine 0.5 mg/kg and famotidine 1.0 mg/kg; second 4 weeks, double-blind withdrawal comparison (safety and efficacy) of each dose with placebo. RESULTS: No serious adverse events were reported. Eleven patients had 16 non-serious, possibly drug-related adverse experiences: 6 patients with agitation or irritability (manifested as head-rubbing in two), 3 patients with somnolence, 2 patients with anorexia, 2 with headache, 1 patient with vomiting, 1 patient with hiccups, and 1 patient with candidiasis. Of the 35 infants, 27 completed Part I. There were significant score improvements for famotidine 0.5 mg/kg in regurgitation frequency (P = 0.04), and for famotidine 1.0 mg/kg in crying time (P = 0.027) and regurgitation frequency (P = 0.004) and volume (P = 0.01). Eight infants completed Part II on double-blind treatment, which was insufficient for meaningful comparisons. CONCLUSIONS: Histamine-2 receptor antagonists may cause agitation and headache in infants. A possibly efficacious famotidine dose for infants is 0.5 mg/kg (frequency adjusted for age). As 1.0 mg/kg may be more efficacious in some, the dosage may require individualization based on response. Further sizeable placebo-controlled evaluations of histamine-2 receptor antagonists in infants with gastro-oesophageal reflux disease are warranted. PMID- 12752347 TI - Rabeprazole is superior to omeprazole for the inhibition of peptone meal stimulated gastric acid secretion in Helicobacter pylori-negative subjects. AB - BACKGROUND: Peptone meal-stimulated gastric acid output is considered to be a reliable means to evaluate drug-mediated inhibition of stimulated gastric acid output, an important measure of the efficacy of the agents--such as proton pump inhibitors--used to treat acid-related disorders. AIM: To compare the initial and overall inhibitory effects on peptone meal-stimulated gastric acid secretion of rabeprazole and omeprazole, 20 mg, in Helicobacter pylori-negative subjects on the first and eighth days of treatment. METHODS: Healthy volunteers (n = 27) were randomized in a single-centre, double-blind, double-dummy, 2 x 2 cross-over study. Subjects received an oral dose of rabeprazole or omeprazole, 20 mg once daily, for 8 days. After a 2-4-week washout period, subjects were crossed over to receive the other medication for 8 days. Peptone meal-stimulated gastric acid secretion was measured at hours 11 and 23 at baseline and on days 1 and 8 of treatment. RESULTS: On days 1 and 8, rabeprazole demonstrated a significantly greater inhibition of peptone meal-stimulated gastric acid secretion compared with omeprazole at all time points (P < 0.03). Median values of steady-state inhibition on day 1 were statistically significant at hour 23 (rabeprazole 100% vs. omeprazole 74%, P < 0.02). CONCLUSIONS: Rabeprazole, 20 mg, demonstrated superior control of peptone meal-stimulated gastric acid secretion compared with omeprazole, 20 mg, after the first dose and after the eighth daily dose. Rabeprazole achieved a more rapid onset of acid inhibition and a greater steady state reduction in peptone meal-stimulated gastric acid secretion. PMID- 12752348 TI - Non-cardiac chest pain: prevalence, risk factors, impact and consulting--a population-based study. AB - BACKGROUND: Little is known about the prevalence and importance of non-cardiac chest pain in the general population. AIM: To evaluate the magnitude and impact of this condition. METHODS: A validated self-report questionnaire was mailed to a sample of 1000 residents of Penrith, selected randomly from the electoral rolls. Symptoms, risk factors, psychological distress, quality of life and demographics were measured. RESULTS: The response rate was 73% (n = 672; mean age, 46 years; 52% female). Chest pain ever was reported by 39% of the population; 7% reported a history of myocardial infarction and 8% a history of angina. Two hundred and nineteen (33%) cases were classified as non-cardiac chest pain; only 23% had consulted a physician about chest pain in the previous year. The only independent risk factor for non-cardiac chest pain was the frequency of heartburn (odds ratio, 1.74; 95% confidence interval, 1.08-2.79; P = 0.02). None of the gastrointestinal (heartburn, dysphagia, acid regurgitation) or psychological (anxiety, depression, neuroticism) risk factors were significantly associated with consulting for non-cardiac chest pain. CONCLUSIONS: Non-cardiac chest pain is remarkably common in the general population and negatively impacts on the quality of life. Gastro-oesophageal reflux disease is a key risk factor for non cardiac chest pain in the community. Health care seeking for non-cardiac chest pain remains unexplained. PMID- 12752349 TI - Helicobacter pylori eradication and gastric ulcer healing--comparison of three pantoprazole-based triple therapies. AB - AIM: To study the efficacy of three pantoprazole-based triple therapy regimens for the eradication of Helicobacter pylori infection and gastric ulcer healing. METHODS: In an open, multi-centre, randomized study, 519 H. pylori-positive patients with active gastric ulcer were randomized to receive pantoprazole (40 mg) (P) and two of three antibiotics: clarithromycin (500 mg) (C), metronidazole (500 mg) (M) or amoxicillin (1000 mg) (A). Triple therapy (PAC, PCM, PAM) was administered twice daily for 7 days, followed by pantoprazole until the ulcer had healed. Antrum and corpus biopsies were taken to determine the pattern of gastritis, to assess the H. pylori status and to determine the strain susceptibility to antibiotics, and from the ulcer margins and base to exclude malignancy. Scores based on the Sydney system were used to categorize the gastritis phenotypically. RESULTS: The H. pylori eradication rates for the per protocol (intention-to-treat) analysis were 89% (67%) for PAC, 83% (68%) for PCM and 76% (60%) for PAM, with a significant difference between PAC and PAM. Healing rates after 4 weeks were 91% for PAM, 90% for PCM and 88% for PAC (per protocol analysis). The eradication rates were lower in patients in whom strains resistant to any antibiotic used in the triple therapies were detected. Successful eradication [odds ratio, 5.2 (3.3; 8.3)] and the ulcer size (< 15 mm) were significant predictors for healing after 4 weeks. The regimens showed a comparable safety profile and compliance. CONCLUSIONS: Pantoprazole-based triple therapies are effective in the eradication of H. pylori infection in gastric ulcer patients, as reported in previous similar sized studies in duodenal ulcer patients. Successful eradication and an ulcer size of < 15 mm are the best predictors of gastric ulcer healing after 4 weeks. PMID- 12752350 TI - Triple vs. quadruple therapy for treating Helicobacter pylori infection: a meta analysis. AB - BACKGROUND: Triple therapy (proton pump inhibitor, clarithromycin and amoxicillin or an imidazole) is the first-line treatment for Helicobacter pylori infection. However, the effectiveness of triple therapy is decreasing due to the increase in antibiotic resistance. Quadruple therapy (proton pump inhibitor, tetracycline, metronidazole and a bismuth salt) is a very effective regimen even in areas of high prevalence of antibiotic resistance, and may be an alternative first-line treatment. AIM: To compare triple vs. quadruple therapy for the first-line treatment of H. pylori infection. METHODS: An extensive literature search was performed to identify randomized trials comparing triple vs. quadruple therapy. Selected trials were included in a meta-analysis using Review Manager 4.1. RESULTS: Four studies met the inclusion criteria. Eradication rates with quadruple therapy were slightly higher in both the intention-to-treat (81% vs. 78%; odds ratio, 0.83; 95% confidence interval, 0.61-1.14) and per protocol (88% vs. 85%; odds ratio, 0.81; 95% confidence interval, 0.55-1.20) analysis, although the differences were not statistically significant. Nor were there significant differences in compliance or adverse effects between the therapies. CONCLUSION: Triple and quadruple therapies seem to be roughly equivalent in terms of effectiveness, compliance and side-effects profile when administered as first line treatment for H. pylori infection. PMID- 12752351 TI - The clinical course of fistulating Crohn's disease. AB - AIM: To determine the clinical characteristics, management and outcome of Crohn's fistulas from the time of first presentation. METHODS: Patients treated for fistulas 6 years previously were assessed for disease demographics, fistula characteristics and treatment from first presentation to final follow-up. RESULTS: Eighty-seven patients with active Crohn's fistulas were evaluated. The median age was 35 years and the median duration of Crohn's disease was 8 years at study entry. Disease was ileo-colonic or colonic in 85%, and 65% had rectal involvement. A single fistula was present in one-third and multiple fistulas in two-thirds; 65% of fistulas were perianal; 80% of fistulas were complex. After a median follow-up from the last treatment of 5.9 years, 68% of patients showed healing of all fistulas, 18% showed healing of some fistulas and 14% showed no healing of fistulas. The fistula site did not influence healing. Perianal and recto-vaginal fistulas took a median of 2.6 years to heal. Half of the complex fistulas required a stoma, resection or proctectomy. CONCLUSIONS: Healing is usually achieved. However, morbidity is great and healing is slow. Proctectomy is required in one-fifth of patients, and perineal healing is often slow. Defining the perianal fistula anatomy as complex or simple determines the likelihood of healing and the type of surgical approach required. PMID- 12752352 TI - Combined scintigraphic and pharmacokinetic investigation of enteric-coated mesalazine micropellets in healthy subjects. AB - BACKGROUND: There is a growing clinical trend to increase the daily dose of mesalazine, which leads to significant compliance issues associated with multiple dosings of current preparations. AIM: To examine the gastrointestinal performance and systemic exposure of a 1.5 g sachet (micropellets) mesalazine formulation, compared with three enteric-coated tablets (500 mg each, Claversal). METHODS: A randomized, two-way, cross-over pharmacoscintigraphic (scintigraphy plus pharmacokinetics) study and a two-way, cross-over, pharmacokinetic-only study were performed in 24 healthy volunteers (12 subjects per investigation). RESULTS: The relative bioavailability of mesalazine was 92% comparing micropellets with Claversal tablets, and the cumulative urine excretion was c. 26% for both preparations, suggesting comparable systemic exposure for the two types of preparation. In the majority of subjects, drug release from the micropellet formulation occurred predominantly in the terminal ileum and ascending colon. The Claversal tablets disintegrated in comparable intestinal sites, albeit at slightly later time points than the micropellets, principally due to slower gastric emptying for the single-unit formulation. CONCLUSION: The 1.5 g micropellet formulation offers comparable delivery properties to the marketed tablets, but with greater convenience of dosing. PMID- 12752353 TI - Gastrointestinal transit and release of 5-aminosalicylic acid from 153Sm-labelled mesalazine pellets vs. tablets in male healthy volunteers. AB - BACKGROUND: Mesalazine (5-aminosalicylic acid)-containing formulations, designed to optimize drug delivery to the ileo-caecal region, represent a cornerstone in the treatment of inflammatory bowel diseases. AIM: : To test, by means of pharmaco-scintigraphy, whether novel mesalazine-containing pellets release 5 aminosalicylic acid in the same target region as mesalazine tablets (Salofalk). METHODS: Fourteen healthy male volunteers received a single dose of either pellets or tablets containing 500 mg of mesalazine and 2 mg of 152Sm2O3 with a 1 week washout period. The gastrointestinal transit of 153Sm, incorporated into the formulations, was followed by gamma-scintigraphy. Mesalazine release was verified by assessing 5-aminosalicylic acid plasma pharmacokinetics. RESULTS: The formulations reached the ileo-caecal target region almost at the same time (3.3 +/- 1 and 3.8 +/- 1 h for pellets and tablets, respectively). Plasma 5 aminosalicylic acid tmax values were comparable and corresponded to the time during which the formulations were located in the target region. Plasma AUC values were significantly lower for pellets, which might be explained by a more prolonged release of 5-aminosalicylic acid. CONCLUSIONS: Novel mesalazine pellets and Salofalk tablets release active 5-aminosalicylic acid in the same target region and pass through the gastrointestinal tract under fasting conditions in healthy volunteers in a comparable time. From a comparison of in vitro dissolution and plasma concentration data, a slower and more prolonged release of 5-aminosalicylic acid from pellets is suggested. PMID- 12752354 TI - Diagnostic investigation rates and use of prescription and non-prescription medications amongst dyspeptics: a population-based study of 2300 Australians. AB - BACKGROUND: : There is limited knowledge of the diagnostic investigation rates and use of prescription and non-prescription drugs amongst dyspeptics. AIM: : To assess the investigation rates and use of prescription and non-prescription anti ulcer medications amongst dyspeptics in the population. METHODS: : A cross sectional survey was performed of 2300 Australians. RESULTS: : Of 748 dyspeptics, 422 (56%) had consulted a doctor regarding dyspepsia at some time in their life. Of the consulters, 64% had undergone investigations at some time: 37% an endoscopy, 54% a barium meal and 27% both. A diagnosis of peptic ulcer was reported by 31% of those investigated. The symptom profile of gastroscopy patients differed significantly from that of uninvestigated dyspeptics. Of the consulters, 36% had taken anti-ulcer prescription drugs in the last 3 months: Histamine-2 receptor antagonists (73% of prescriptions), proton pump inhibitors (17%), cytoprotectants (5%) and prokinetic drugs (5%). Antacids were taken by 30% of non-consulting dyspeptics, 44% of consulters not on prescription drugs and 58% of dyspeptics taking prescription drugs. Advancing age, but not gender, was associated with diagnostic investigation and prescription and non-prescription drug usage. CONCLUSIONS: : There are high rates of diagnostic investigation amongst dyspeptics who consult doctors. Many individuals with dyspepsia decide to self-medicate with antacids regardless of consulting or prescriptions, suggesting that current management is suboptimal. PMID- 12752355 TI - Development and validation of the Diabetes Bowel Symptom Questionnaire. AB - BACKGROUND: Although gastrointestinal symptoms occur frequently, there is no validated measure of gastrointestinal symptoms in patients with diabetes mellitus. AIM: To develop the Diabetes Bowel Symptom Questionnaire. METHODS: A questionnaire derived from previously validated symptom measures was compiled to assess all relevant gastrointestinal and diabetes items. Face and content validity were ascertained by expert review. One hundred and sixty-eight patients with diabetes mellitus completed the instrument, and reliability was evaluated by a test-re-test procedure 1 week later. Concurrent validity was evaluated by an independent physician interview (n = 33). Measures of glycaemic control (glycated haemoglobin and plasma glucose) were compared with self-reported glycaemic control on a five-point Likert scale in diabetic out-patients (n = 166). RESULTS: The questionnaire had adequate face and content validity. There was good to excellent test-re-test reliability for the gastrointestinal and diabetes items (median kappa: 0.63 and 0.79, respectively); concurrent validity was good to excellent (median kappa: 0.47 and 0.65, respectively), except for the items assessing the severity of gastrointestinal symptoms. Both glycated haemoglobin (P < 0.0001) and plasma glucose (P = 0.005) correlated significantly with self reported glycaemic control. CONCLUSION: The Diabetes Bowel Symptom Questionnaire appears to be a useful measure of gastrointestinal symptoms and glycaemic control in diabetes mellitus, and should have applicability in epidemiological and clinical studies. PMID- 12752356 TI - Diverticular disease of the colon--on the rise: a study of hospital admissions in England between 1989/1990 and 1999/2000. AB - BACKGROUND: Diverticular disease emerged as a common problem in Western countries over the course of the 20th century. AIMS: To determine the time trends in diverticular disease for hospital admissions in England between 1989/1990 and 1999/2000 and population mortality rates from 1979 to 1999. METHODS: Hospital Episode Statistics were obtained from the Department of Health and mortality data from the Office for National Statistics. RESULTS: Between 1989/1990 and 1999/2000, annual age-standardized hospital admission rates for diverticular disease increased by 16% for males (from 20.1 to 23.2 per 100,000) and 12% for females (from 28.6 to 31.9 per 100,000). Female rates were significantly higher than male rates throughout the study period. The proportions of admissions with an operation increased by 16% for males (from 22.9% to 24.1%) and 14% for females (from 19.7% to 22.3%). Older patients were less likely to undergo operation than younger patients. In-patient case fatality rates and population mortality rates remained unchanged. CONCLUSIONS: Admission rates for diverticular disease increased over the study period. The proportion of patients who underwent operation increased, but in-patient and population mortality rates remained unchanged. With an ageing population, diverticular disease will become an increasingly important clinical problem in England. PMID- 12752357 TI - Induction interferon and ribavirin for re-treatment of chronic hepatitis C patients unresponsive to interferon alone. AB - BACKGROUND: The optimal treatment for hepatitis C patients unresponsive to interferon is unclear. High-dose induction interferon may enhance early viral clearance, whilst ribavirin reduces relapse; in combination, they may improve sustained virological response rates. AIM: To compare the efficacy and safety of re-treatment with interferon induction, with or without ribavirin, in interferon non-responders. METHODS: We randomized 218 biochemical interferon non-responders to 10 MU interferon alpha 2b daily for 4 weeks, followed by 5 MU thrice weekly for 48 weeks plus ribavirin (II + R), or to the same interferon regimen plus placebo (II + P). All patients were viraemic at entry. RESULTS: The sustained virological response in the II + R group was 39%[95% confidence interval (CI), 30 48%], compared with 16% (95% CI, 9-23%) in the II + P group (P < 0.002). The study drug was discontinued for intolerable symptoms during induction in 9% of the II + R group and in 5% of the II + P group. By logistic regression, a sustained virological response was more likely following II + R treatment (odds ratio, 4.4; 95% CI, 2.1-9.7) and less likely in patients with genotype 1 or 4 (odds ratio, 0.16; 95% CI, 0.07-0.36). CONCLUSION: High-dose induction interferon plus ribavirin is well tolerated and effective for patients unresponsive to interferon alone. PMID- 12752359 TI - Familial influences on adolescent smoking. AB - The family unit is the primary source of transmission of basic social, cultural, genetic, and biological factors that may underlie individual differences in smoking. Existing information on the role of familial factors in tobacco use is characterized by two separate, but somewhat overlapping, lines of research: genetic epidemiological studies and risk-factor research. The present paper summarizes and evaluates studies assessing the association between adolescent smoking and parent and sibling smoking behaviors. A review of 87 studies reveals that methods are limited by a lack of standardized instruments, failure to measure important confounding and mediating factors, reliance on cross-sectional designs and the use of inconsistent definitions of tobacco-related behavior and assessment procedures. Moreover, there are no systematic family studies of the acquisition and continuation of smoking that have employed contemporary methodological standards for examining familial aggregation of tobacco behaviors among adolescents. Findings across studies show weak and inconsistent associations between parent and adolescent smoking; inconsistent findings may be attributed to methodological issues or associated factors that may complicate the relation between parent and adolescent smoking. Sibling and peer smoking show greater associations with adolescent smoking. Suggestions for future research include contemporary family studies that delineate meaningful phenotypes of tobacco use and prospective work on the later stages of tobacco use and the timing of the influence and valence of parent and family factors. Integration of the risk factor approach within the family study design may enrich both approaches to elucidate familial influences on smoking. PMID- 12752360 TI - Theory, measurement, and methods in the study of family influences on adolescent smoking. AB - This paper discusses three dilemmas faced by researchers interested in family influences in substance use: the transitional nature of adolescent smoking, the complexity and multi-dimensionality of family forms and influences, and the inter relationship of family influences with other key developmental contexts. Methodological and conceptual issues stemming from these dilemmas are discussed with regard to understanding why previous reviews have found the correlations between family predictors and adolescent smoking to be relatively low. In particular, the importance of understanding time, the transitional nature of the phenomenon, and within- and between- family processes are emphasized. More appropriate conceptual and statistical models for analyzing family influences are suggested, including both mixed models and person-centered approaches. PMID- 12752361 TI - Peers and adolescent smoking. AB - There is a considerable body of empirical research that has identified adolescent peer relationships as a primary factor involved in adolescent cigarette smoking. Despite this large research base, many questions remain unanswered about the mechanisms by which peers affect youths' smoking behavior. Understanding these processes of influence is key to the development of prevention and intervention programs designed to address adolescent smoking as a significant public health concern. In this paper, theoretical frameworks and empirical findings are reviewed critically which inform the current state of knowledge regarding peer influences on teenage smoking. Specifically, social learning theory, primary socialization theory, social identity theory and social network theory are discussed. Empirical findings regarding peer influence and selection, as well as multiple reference points in adolescent friendships, including best friendships, romantic relationships, peer groups and social crowds, are also reviewed. Review of this work reveals the contribution that peers have in adolescents' use of tobacco, in some cases promoting use, and in other cases deterring it. This review also suggests that peer influences on smoking are more subtle than commonly thought and need to be examined more carefully, including consideration of larger social contexts, e.g. the family, neighborhood, and media. Recommendations for future investigations are made, as well as suggestions for specific methodological approaches that offer promise for advancing our knowledge of the contribution of peers on adolescent tobacco use. PMID- 12752362 TI - An ecological approach to understanding youth smoking trajectories: problems and prospects. AB - Non-random patterns of aggregate youth smoking rates and trajectories across communities suggest that individual-level characteristics cannot account fully for the behavior in question. Instead, at least part of the explanation must lie somewhere within the community context. Such community-level contextual effects can impact directly both group and individual-level behavior (e.g. main effects), and they can also condition the effects of individual-level factors on individual behaviors (e.g. moderating effects). This paper reviews previous research examining community-level contextual effects regarding smoking and substance use more generally and identifies important limitations of this extant work, thus defining an agenda for future empirical studies. Next, the (in)compatibility of previous empirical findings with current theoretical models is discussed. In offering an alternative to these existing models, the paper concludes with presentation and discussion of a multi-level, integrated model of adolescent smoking trajectories. In this model, community/institutional forces are presumed to impact smoking above and beyond individual-level main effects. These posited community-level forces are broad and varied, representing school characteristics, neighborhood demographic characteristics, religious culture, media influence, economic context, health services and so on. In addition to exhibiting contextual main effects, the effects of community in the proposed multilevel model can be mediated by community-level processes, including the processes of control and socialization discussed herein. Also, community-level characteristics may interact in producing certain tobacco-use outcomes and, perhaps most importantly, they may moderate or condition the effects of interindividual differences on smoking. PMID- 12752363 TI - Role of the media in influencing trajectories of youth smoking. AB - This paper summarizes results of empirical studies on cigarette advertising and promotions, antismoking advertising, product placement in movies, on television and in music media and news coverage about smoking. In addition, we provide an overview of some of the theoretical literature relevant to the study of media uses and effects. Finally, we discuss empirical findings in the context of these theories to draw some conclusions about media influences on smoking and identify issues for further research. We conclude that (a) the media both shape and reflect social values about smoking; (b) the media provide new information about smoking directly to audiences; (c) the media act as a source of observational learning by providing models which teenagers may seek to emulate; (d) exposure to media messages about smoking also provides direct reinforcement for smoking or not smoking; (e) the media promote interpersonal discussion about smoking; (f) the media can influence "intervening" behaviors that may make teenage smoking less likely; and (g) antismoking media messages can also set the agenda for other change at the community, state or national level. We outline priorities for further research which emphasize the need for longitudinal studies, multi-level studies, an awareness of the probably dynamic relationship between tobacco advertising and antismoking advertising, the importance of determining appraisal of tobacco industry youth smoking prevention efforts and the dearth of research on news coverage about smoking. PMID- 12752364 TI - Prices, policies and youth smoking, May 2001. AB - Youth smoking continues to be an important public health problem. Many policy tools designed to reduce youth smoking are based on economic principles. This paper describes the economic and policy context of tobacco use aiming at reducing youth smoking and explains the economic rationale for tobacco control tools such as excise tax and price, clean indoor air laws, youth access laws and the broad provision of health information to the public. An overview of economic models of addiction provides the framework for empirical analysis of the impact of these policies. This is followed by a summary of the empirical evidence of the effectiveness of various tobacco control tools that are primarily economic in nature. The most consistent finding in this literature is that higher cigarette prices discourage youth smoking. Compared to the effects of cigarette taxation and price on youth smoking, the evidence on the effectiveness of the youth access laws and clean indoor air laws are still mixed and inconclusive. More research is needed to address issues such as: (1) the effects of gender, age, race and socio economic status on the relationship between tobacco control policies and youth smoking; (2) better measurement of the outcome variables to account for the multi dimensional nature of dependence; and (3) the effects of excise taxes and other tobacco control policies with regard to a host of dimensions of smoking such as initiation, cessation, and more generally the trajectories of tobacco use that would include patterns of progression, maintenance, regression, cessation, and relapse. More frequently collected longitudinal data than those currently available are needed to address the above issues. Understanding smoking behavior cannot be achieved without incorporating familial and other social contexts. PMID- 12752365 TI - Political economy of youth smoking regulation. AB - This paper considers the political economy of tobacco regulation-that is, the interplay between politics and markets in tobacco regulation. In surveying the interest-group politics of tobacco control, it describes political advantages held by a profitable and concentrated industry engaged in tobacco production. It also considers reasons for the surprising political success of tobacco control advocates over the past decade. The paper considers challenges faced by tobacco control advocates, who face their own obstacles to collective action in promoting effective tobacco regulation. Both public officials and tobacco control advocates face stronger incentives to advocate new legislation than they do to ensure effective implementation of existing regulatory measures. The essay concludes by considering the Master Settlement Agreement, whose detailed implementation remains uncertain after the election of the Bush administration. PMID- 12752366 TI - Smoking: what does culture have to do with it? AB - In this commentary issues are raised relating to the role of ethnicity and "culture" as a context influencing adolescent smoking. A processual rendering of culture is encouraged, as is an appreciation of intraethnic diversity. The question posed is "what is cultural about particular patterns, transitions and trajectories of smoking?" Productive ways of investigating patterns of smoking which attend to class, ethnicity, gender norms, modernity and popular culture are focused upon as an ongoing project subject to both the identity needs of youth and the agenda of the tobacco industry. Promising areas of research are identified, as are the potential contributions of ethnographies of tobacco use. PMID- 12752367 TI - Contextual factors and youth tobacco use: policy linkages. AB - This paper provides a short commentary on the set of papers contained in this special issue that discuss various contextual factors that affect youth smoking. It highlights the interrelationships between the economic and policy factors, media influences, community factors, peer influences and familial factors that impact on youth smoking. Particular emphasis is given to the direct effects of prices and policies on youth smoking, and to the indirect effects of these factors as they work through the other contextual factors. PMID- 12752368 TI - The case for studying multiple contexts simultaneously. AB - Individuals live in multiple social contexts simultaneously, whether they be family, peer, neighborhood, school or work contexts. In individual lives, it is likely that the forces within any one context that promote healthy human development are correlated with whatever causal forces operate to the same end in other contexts. This paper explores why it is important to consider social contexts, not singly as in traditional neighborhood or school or work-site research, but in combination. PMID- 12752370 TI - Computer model of passive signal integration based on whole-cell in vitro studies of rat lateral geniculate nucleus. AB - Computer models were used to investigate passive properties of lateral geniculate nucleus thalamocortical cells and thalamic interneurons based on in vitro whole cell study. Two neurons of each type were characterized physiologically and morphologically. Thalamocortical cells transmitted 37% of steady-state signal orthodromically (distal dendrite to soma) and 93% antidromically (soma to distal dendrite); interneurons transmitted 18% orthodromically and 53% antidromically. Lowering membrane resistance caused a dramatic drop in steady-state signal transmission. Simulation of brief signals such as orthodromically transmitted postsynaptic potentials and antidromically transmitted action potentials showed relatively poor transmission due to the low-pass filtering property of dendrites. This attenuation was particularly pronounced in interneurons. By contrast, bursts of postsynaptic potentials or action potentials were relatively well transmitted as the temporal summation of these recurring signals gave prolonged depolarizations comparable to prolonged current injection. While synaptic clustering, active channels and reduction of membrane resistance by ongoing synaptic activity will have additional profound effects in vivo, the present in vitro modelling suggests that passive signal transmission in neurons will depend on type of signal conveyed, on directionality and on membrane state. This will be particularly important for thalamic interneurons, whose presynaptic dendrites may either work independently or function in concert with each other and with the soma. Our findings suggest that bursts may be particularly well transmitted along dendrites, allowing firing format to alter the functional anatomy of the cell. PMID- 12752371 TI - Differential expression and cellular localization of doublecortin in the developing rat retina. AB - Doublecortin is 40 kDa microtubule-associated phosphoprotein required for neuronal migration and differentiation in various regions of the developing central nervous system. We have investigated the expression and cellular localization of doublecortin in the developing rat retina using immunocytochemistry and Western blot analysis. The expression of doublecortin was high from embryonic day 18 (E18) until E20 and was low during the postnatal period. The doublecortin immunoreactivity first appeared in a few radially orientated cells in the mantle zone of the primitive retina at E15. From E16 onward, the immunoreactivity appeared in two different regions: the inner part of the retina and middle of the neuroblastic layer. In the inner part, the somata of cells in the ganglion cell layer, in the distal row of the neuroblastic layer and profiles in the inner plexiform layer showed doublecortin immunoreactivity up to postnatal day 1 (P1). Afterwards, the doublecortin immunoreactivity persisted in the inner plexiform layer until P15, although the intensity decreased gradually with the maturation of the retina. In the middle of the neuroblastic layer, doublecortin immunoreactivity appeared in the radially orientated cells. These cells transformed into horizontal cells. The doublecortin immunoreactivity persisted in these cells up to P21. Given these results, doublecortin may play an important role in the migration and differentiation of specific neuronal populations in developmental stages of the rat retina. PMID- 12752372 TI - Mineralocorticoid receptor expression and increased survival following neuronal injury. AB - Glucocorticoids, acting via the mineralocorticoid receptor, are required for granule neuronal survival in the rat dentate gyrus. Whether this mineralocorticoid receptor-mediated neuroprotective effect has more general applicability is unknown. Here we report increased mineralocorticoid receptor expression in rat hippocampal and cortical neurons exposed in vitro to low levels of staurosporine and in rat hippocampal pyramidal neurons exposed in vivo to hypothermic transient global ischaemia. In both the cell culture system and the in vivo system increased mineralocorticoid receptor expression is associated with increased neuronal survival, and this increase is reversed by mineralocorticoid receptor antagonism. Modulation of mineralocorticoid receptor gene expression may therefore be an important target for reduction of brain injury in conditions caused by cerebral ischaemia including brain damage following cardiac arrest and stroke. PMID- 12752373 TI - Blockade of cortical muscarinic but not NMDA receptors prevents a novel taste from becoming familiar. AB - Exposure to a novel taste solution in the rat is followed by a decrease in its intake known as neophobia. This effect gradually disappears, and consumption increases from the second presentation of the taste (attenuation of neophobia), reflecting that the animal learned that it is safe to drink it. Conversely, if gastric malaise is induced after first intake, the rat will develop a long lasting aversion (conditioned taste aversion). Previous attempts to elucidate the physiological nature of taste memory trace stems only from procedures that require malaise to measure taste memory. Here we assess the relevance of both muscarinic and N-methyl-d-aspartate receptors, known to be involved in conditioned taste aversion, on taste memory using a nonaversive procedure (attenuation of neophobia learning). Attenuation of neophobia was impaired by the muscarinic receptor antagonist, scopolamine, microinjected 20 min before, immediately after or up to 2 h after the first taste experience, suggesting that muscarinic receptors are involved in the acquisition and consolidation of attenuation of neophobia learning. However, the N-methyl-d-aspartate receptor antagonist, d,l-2-amino-5-phosphonovaleric acid, did not affect attenuation of neophobia even when the same dose of the drug was able to disrupt conditioned taste aversion learning, which suggests that attenuation of neophobia learning would be independent of N-methyl-d-aspartate receptors activity in the insular cortex. The neophobic response induced by strong saccharin presentation was not affected by either of the treatments given, which rules out any impairment in taste perception. These results indicate that while cortical muscarinic receptors are important in the formation and consolidation of safe memory trace, N-methyl-d aspartate receptor activity appears to be noncritical. PMID- 12752374 TI - The ability of atypical antipsychotic drugs vs. haloperidol to protect PC12 cells against MPP+-induced apoptosis. AB - The present study examined the effects of the atypical antipsychotic drugs clozapine, olanzapine, quetiapine and risperidone, on N-methyl-4-phenylpyridinium ion-induced apoptosis and DNA damage in PC12 cells, and explored the molecular mechanisms underlying these effects. Haloperidol, a typical antipsychotic drug, was used for comparison. Exposure of PC12 cells to 50 micro m N-methyl-4 phenylpyridinium ion for 24 h resulted in a 35-45% loss of cells in culture. Pretreatment with the aforementioned atypical antipsychotic drugs significantly reduced the N-methyl-4-phenylpyridinium ion-induced cell loss, whereas haloperidol (10-100 micro m) did not have this protective effect. Hoechst 33258 staining revealed the apoptotic nuclear features of the N-methyl-4 phenylpyridinium ion-induced cell death, and showed that the atypical antipsychotic drugs, but not haloperidol, effectively prevented PC12 cells from this N-methyl-4-phenylpyridinium ion-induced apoptosis. DNA fragmentation assays further confirmed the N-methyl-4-phenylpyridinium ion-induced nuclear fragmentation. Pretreatment with the atypical antipsychotic drugs completely prevented this nuclear fragmentation, whereas haloperidol only partially prevented it. In vitro oligonucleotide assays indicated an activation of a specific glycosylase that recognizes and cleaves bases (at the 8-hydroxyl-2 deoxyguanine site) that were damaged by N-methyl-4-phenylpyridinium ion. Pretreatment with the atypical antipsychotic drugs more effectively attenuated this N-methyl-4-phenylpyridinium ion-induced activation than did haloperidol. Northern blot analyses showed that the atypical antipsychotic drugs, but not haloperidol, blocked the N-methyl-4-phenylpyridinium ion-induced substantial increase of copper/zinc superoxide dismutase mRNA in PC12 cells. Atypical antipsychotic drugs slightly up-regulated the expression of copper/zinc superoxide dismutase mRNA, whereas haloperidol strongly increased the expression of copper/zinc superoxide dismutase mRNA. These data may account for the different therapeutic effects and side-effect profiles of typical and atypical antipsychotic drugs in schizophrenia. PMID- 12752375 TI - Neuronal lineage-specific induction of phospholipase Cepsilon expression in the developing mouse brain. AB - Phospholipase C is a key enzyme of intracellular signal transduction in the central nervous system. We and others recently discovered a novel class of phospholipase C, phospholipase Cepsilon, which is regulated by Ras and Rap small GTPases. As a first step toward analysis of its function, we have examined the spatial and temporal expression patterns of phospholipase Cepsilon during mouse development by in situ hybridization and immunohistochemistry. Around embryonic day 10.5, abundant expression of phospholipase Cepsilon is observed specifically in the outermost layer of the neural tube. On embryonic day 12 and later, it is observed mainly in the marginal zone of developing brain and spinal cord as well as in other regions undergoing neuronal differentiation, such as the retina and olfactory epithelium. The phospholipase Cepsilon-expressing cells almost invariably express microtubule-associated protein 2, but hardly express nestin or glial fibrillary acidic protein, indicating that the expression of phospholipase Cepsilon is induced specifically in cells committed to the neuronal lineage. The expression of phospholipase Cepsilon persists in the terminally differentiated neurons and exhibits no regional specificity. Further, an in vitro culture system of neuroepithelial stem cells is employed to show that abundant expression of phospholipase Cepsilon occurs in parallel with the loss of nestin expression as well as with the induction of microtubule-associated protein 2 expression and neuronal morphology. Also, glial fibrillary acidic protein-positive glial lineage cells do not exhibit the high phospholipase Cepsilon expression. These results suggest that the induction of phospholipase Cepsilon expression may be a specific event associated with the commitment of the neural precursor cells to the neuronal lineage. PMID- 12752376 TI - A hot spot for hotfoot mutations in the gene encoding the delta2 glutamate receptor. AB - The orphan glutamate receptor delta2 is selectively expressed in Purkinje cells and plays a crucial role in cerebellar functions. Recently, ataxia in the hotfoot mouse ho4J was demonstrated to be caused by a deletion in the delta2 receptor gene (Grid2) removing the N-terminal 170 amino acids of the delta2 receptor. To understand how delta2 receptors function, we characterized mutations in eight additional spontaneously occurring hotfoot alleles of Grid2. The mouse Grid2 gene consists of 16 exons, spanning approximately 1.4 Mb. Genomic DNA analysis showed that seven hotfoot mutants had a deletion of one or more exons encoding the N terminal domain of delta2 receptors. The exception is ho5J, which has a point mutation in exon 12. Deletions in ho7J, ho9J, ho11J and ho12J mice result in the in-frame deletion of between 40 and 95 amino acids. Expression of constructs containing these deletions in HEK293 cells resulted in protein retention in the endoplasmic reticulum or cis-Golgi without transport to the cell surface. Coimmunoprecipitation assays indicated that these deletions also reduce the intermolecular interaction between individual delta2 receptors. These results indicate that the deleted N-terminal regions are crucial for oligomerization of delta2 receptors and their subsequent transport to the cell surface of Purkinje cells. The relatively large size of the Grid2 gene may be one of the reasons why many spontaneous mutations occur in this gene. In addition, the frequent occurrence of in-frame deletions within the N-terminal domain in hotfoot mutants suggests the importance of this domain in the function of delta2 receptors. PMID- 12752377 TI - Calcium-dependent phosphorylation processes control brain aromatase in quail. AB - Increased gene transcription activated by the binding of sex steroids to their cognate receptors is one important way in which oestrogen synthase (aromatase) activity is regulated in the brain. This control mechanism is relatively slow (hours to days) but recent data indicate that aromatase activity in quail preoptic-hypothalamic homogenates is also rapidly (within minutes) affected by exposure to conditions that enhance Ca2+-dependent protein phosphorylation. We demonstrate here that Ca2+-dependent phosphorylations controlled by the activity of multiple protein kinases including PKC, and possibly also PKA and CAMK, can rapidly down-regulate aromatase activity in brain homogenates. These phosphorylations directly affect the aromatase molecule itself. Western blotting experiments on aromatase purified by immunoprecipitation reveal the presence on the enzyme of phosphorylated serine, threonine and tyrosine residues in concentrations that are increased by phosphorylating conditions. Cloning and sequencing of the quail aromatase identified a 1541-bp open reading frame that encodes a predicted 490-amino-acid protein containing all the functional domains that have been previously described in the mammalian and avian aromatase. Fifteen predicted consensus phosphorylation sites were identified in this sequence, but only two of these (threonine 455 and 486) match the consensus sequences corresponding to the protein kinases that were shown to affect aromatase activity during the pharmacological experiments (i.e. PKC and PKA). This suggests that the phosphorylation of one or both of these residues represents the mechanism underlying, at least in part, the rapid changes in aromatase activity. PMID- 12752379 TI - The ERK/MAP kinase pathway couples light to immediate-early gene expression in the suprachiasmatic nucleus. AB - Signalling via the p42/44 mitogen-activated protein kinase (MAPK) pathway has been identified as an intermediate event coupling light to entrainment of the mammalian circadian clock located in the suprachiasmatic nucleus (SCN). Given this observation, it was of interest to determine where within the entrainment process the MAPK pathway was functioning. In this study, we examined the role of the MAPK pathway as a regulator of light-induced gene expression in the SCN. Towards this end, we characterized the effect pharmacological disruption of the MAPK cascade has on the expression of the immediate-early genes c-Fos, JunB and EGR-1. We report that uncoupling light from MAPK pathway activation attenuated the expression of all three gene products. In the absence of photic stimulation, inhibition of the MAPK pathway did not alter basal gene product expression levels. Light-induced activation of cAMP response element (CRE)-dependent transcription, as assessed using a CRE-LacZ transgenic mouse strain, was also disrupted by blocking MAPK pathway activation. These results reveal that the MAPK cascade functions as one of the first transduction steps leading from light to rapid transcriptional activation, an essential event in the entrainment process. MAPK pathway-dependent gene expression in the SCN may result, in part, from stimulation of CRE-dependent transcription. PMID- 12752378 TI - Dentate granule cell GABA(A) receptors in epileptic hippocampus: enhanced synaptic efficacy and altered pharmacology. AB - The dentate gyrus (DG) normally functions as a filter, preventing propagation of synchronized activity into the seizure-prone hippocampus. This filter or 'gatekeeper' attribute of the DG is compromised in various pathological states, including temporal lobe epilepsy (TLE). This study examines the role that altered inhibition may play in the deterioration of this crucial DG function. Using the pilocarpine animal model of TLE, we demonstrate that inhibitory synaptic function is altered in principal cells of the DG. Spontaneous miniature inhibitory postsynaptic currents (mIPSCs) recorded in dentate granule cells (DGCs) from epileptic animals were larger, more sensitive to blockade by zinc and less sensitive to augmentation by the benzodiazepine type site 1 modulator zolpidem. Furthermore, mIPSCs examined during a quiescent period following injury but preceding onset of epilepsy were significantly smaller than those present either in control or in TLE DGCs, and had already acquired sensitivity to blockade by zinc prior to the onset of spontaneous seizures. Rapid agonist application experiments demonstrated that prolonged (>35 ms) exposure to zinc is required to block GABAA receptors (GABAARs) in patches pulled from epileptic DGCs. Therefore, zinc must be tonically present to block DGC GABAARs and alter DG function. This would occur only during repetitive activation of mossy fibres. Thus, in the pilocarpine animal model of TLE, an early, de novo, expression of zinc-sensitive GABAARs is coupled with delayed, epilepsy-induced development of a zinc delivery system provided by aberrant sprouting of zinc-containing mossy fibre recurrent collaterals. The temporal and spatial juxtaposition of these pathophysiological alterations may compromise normal 'gatekeeper' function of the DG through dynamic zinc-induced failure of inhibition, predisposing the hippocampal circuit to generate seizures. PMID- 12752380 TI - Periodic absence of nursing mothers phase-shifts circadian rhythms of clock genes in the suprachiasmatic nucleus of rat pups. AB - Effects of absence of nursing mothers on the circadian pacemaker of their offspring were examined by measuring clock genes, the rat Per1 (rPer1) and rPer2 expression rhythms in the pup suprachiasmatic nuclei (SCN). Neonate rats born to mothers kept under a 12-h light : 12-h dark cycle (LD) were blinded immediately after birth and exposed to periodic maternal deprivation where pups were deprived of their mothers during the light phase of 12-h for the first week of life. At postnatal day 6, the periodic maternal deprivation completely phase-reversed the circadian rhythms in expression of the clock genes in the pup SCN and in spontaneous locomotor activity after the pups were weaned at postnatal day 21. The periodic maternal absence also altered the patterns of stress-related gene expressions such as corticotropine-releasing hormone, arginine vasopressin, and glucocorticoid receptor in particular brain areas of the mother-deprived pups at P6. These findings indicate that periodic absence of the nursing mother in the first week of life produces a resetting effect on the neonatal circadian clock and induces stress responses in the hypothalamus-pituitary-adrenal axis. PMID- 12752381 TI - Inhibition of rostral basal forebrain neurons promotes wakefulness and induces FOS in orexin neurons. AB - The present study examined whether the activities of the rostral basal forebrain neurons alter the activities of the orexin (also known as hypocretin) neurons in the tuberal part of the hypothalamus in rats. We performed microdialysis perfusion of the ventromedial portion of the rostral basal forebrain with the GABAA receptor agonist muscimol to inhibit focally the neuronal activities in the rostral basal forebrain. Then, we monitored sleep/wake behaviour and investigated the pattern of activities of orexin neurons by examining the expression of FOS as an indicator of cellular activation. Bilateral perfusion with muscimol (5, 15, and 50 micro m) produced a dose-dependent decrease in the amount of sleep. This perfusion with muscimol at 50 micro m produced FOS-like immunoreactivity in 37% of the orexin neurons located in the tuberal part of the hypothalamus, whereas the FOS-like immunoreactivity was sparse in orexin neurons of the sleeping control rats (P = 0.001 by Mann-Whitney U-test). Unilateral perfusion with muscimol (50 micro m) also suppressed sleep. In this case, FOS-like immunoreactivity was seen in 40% of the orexin neurons on the side ipsilateral to the perfusion site but only in 10% of orexin neurons on the contralateral side (P = 0.018 by Wilcoxon signed rank test). These functional data suggested that a sleep-generating element in the ventromedial part of the rostral basal forebrain provides an inhibitory influence on the activities of the orexin neurons in the tuberal part of the hypothalamus. PMID- 12752383 TI - Differential behavioural sensitization to intermittent morphine treatment in alcohol-preferring AA and alcohol-avoiding ANA rats: role of mesolimbic dopamine. AB - Alcohol-preferring AA (Alko Alcohol) and alcohol-avoiding ANA (Alko Non-Alcohol) rats have well-documented differences in their voluntary ethanol consumption and brain opioidergic systems. The aim of the present study was to investigate whether these rat lines differ in their susceptibility to morphine-induced behavioural and neurochemical sensitization. The rats were given 15 injections of morphine (10 mg/kg, s.c.) or saline every other day. Locomotor activity and release of dopamine in the nucleus accumbens were monitored after a challenge with additional morphine injections (10 mg/kg) 1 and 5 weeks after withdrawal from the repeated treatment. Morphine increased locomotion more in the previously morphine-treated rats than in the saline-treated controls. Furthermore, AA rats were more sensitive to this effect of morphine than ANA rats. Accumbal morphine induced dopamine release was significantly higher in the morphine-treated AA than ANA rats after the first challenge injection 1 week from withdrawal, but no differences were observed after the second challenge. The brain and plasma concentrations of morphine were similar among the lines suggesting that the differences in the effects of morphine cannot be explained in terms of differential pharmacokinetics of morphine in these lines. These data show that AA rats are more susceptible to morphine-induced behavioural sensitization than ANA rats. Furthermore, it suggests that mesolimbic dopamine has at best only a transient role in the expression of opioid-induced behavioural sensitization. The relationship between the mechanisms underlying the differential sensitivity of these rat lines to the effects of repeated morphine and voluntary ethanol drinking remains to be determined. PMID- 12752382 TI - The transient depression of hippocampal CA1 LTP induced by chronic intermittent ethanol exposure is associated with an inhibition of the MAP kinase pathway. AB - Using electrophysiological and biochemical approaches, we investigated the effects of chronic, intermittent ethanol (CIE) treatment on activation of the mitogen activated protein kinase (MAPK), also known as extracellular signal regulated protein kinase 1 and 2. In hippocampal slices taken from control rats, brief high-frequency stimulation to Schaffer collateral fibers induced a large post-tetanic potentiation (PTP) in the CA1 region that decayed to stable long term potentiation (LTP) of field extracellular postsynaptic potentials. Western blot analyses showed that phosphorylation of MAPK was increased during PTP and returned to baseline levels during LTP. In slices from the rats removed immediately from CIE treatment, PTP and MAPK activation during the PTP was significantly less than that observed in control slices and LTP was absent. In slices from rats subjected to 1 day withdrawal from CIE treatment, both the reduction in MAPK phosphorylation during PTP and the impairment of PTP and LTP were still evident. Recovery of PTP and partial recovery of LTP was observed in slices obtained from 5-day withdrawn rats. However, MAPK activation during PTP was still attenuated significantly. Interestingly, MAPK activation was enhanced significantly during LTP in 5-day withdrawn rats as well as the sensitivity to MAPK inhibitor PD 098059. In addition to these changes in HFS-induced MAPK activation, we also observed a significant reduction in the basal phosphorylation of MAPK in slices removed from rats immediately after CIE treatment. These results implicate the MAPK signal transduction pathway as a potential cellular target of ethanol. Alterations in MAPKs could play an important role in the alcohol-induced changes in synaptic plasticity associated with the effects of alcohol abuse on learning and memory processes. PMID- 12752384 TI - Aversive conditioning following repeated withdrawal from ethanol and epileptic kindling. AB - Repeated withdrawal from ethanol, a procedure which resembles amygdala kindling in increasing seizure sensitivity, impairs the acquisition of fear conditioning (Stephens et al., 2001, Eur. J. Neurosci.,14, 2023-31). In contrast, rats previously kindled by repeated electrical stimulation of basolateral amygdala, or repeated administration of pentylenetetrazol, showed increased suppression of operant responding during the presentation of a stimulus conditioned to footshock when conditioning took place several weeks following the kindling experience. Neither form of kindling nor repeated ethanol withdrawal altered taste aversion conditioning, though rats treated chronically with ethanol and given a single withdrawal experience showed enhanced taste aversion conditioning. These results suggest that, despite evidence suggesting a common neuronal mechanism underlying seizure sensitivity following these types of kindling, they differ in their effects on fear conditioning. PMID- 12752385 TI - Stimulation site determines the conditioned effects of kindling in rats: anterior neocortex versus amygdala. AB - Rats received 53 stimulations to either the left basolateral amygdala (BA) or left anterior neocortex (AN) in one environment (CS+) and 53 sham stimulations (the stimulation lead was attached but no current was delivered) in another environment (CS-), quasirandomly over 54 days. Confirming a previous report [Barnes, S.J., Pinel, J.P., Francis, L.H. & Wig, G.S. (2001) Behav. Neurosci., 115, 1065-1072], as BA kindling progressed, the CS+ began to elicit more defensive behaviours (i.e. less activity, more freezing and avoidance of the CS+) than the CS-, and at the end of the experiment, convulsions elicited in the CS+ were more severe than those elicited in the CS-. Like BA kindling, AN kindling led to less activity in the CS+; but unlike BA kindling, AN kindling led to more wet-dog-shakes and less, rather than more, severe convulsions in the CS+. During AN kindling, the mean number of wet-dog-shakes in the CS+ was negatively correlated with the mean convulsion class, suggesting that wet-dog-shakes contribute to the inherent variability of AN kindling. These findings confirm that inherent conditioned effects influence kindled convulsions and interictal behaviour and establish for the first time that the pattern of these conditioned effects is a function of the kindling site. PMID- 12752386 TI - Double dissociation of the effects of lesions of basolateral and central amygdala on conditioned stimulus-potentiated feeding and Pavlovian-instrumental transfer. AB - Pavlovian conditioned stimuli (CSs) for food can enhance both the performance of instrumental responses that earn food (Pavlovian-instrumental transfer; PIT) and the consumption of food itself (CS-potentiated feeding). After a single phase of Pavlovian training, each rat was tested in both PIT and potentiated feeding tasks. Rats with lesions of the central nucleus of the amygdala failed to exhibit PIT but showed normal CS-potentiated feeding. By contrast, rats with lesions of the basolateral amygdala showed normal PIT but failed to display CS-potentiated feeding. Performances in a variety of comparison conditions suggested that both lesion effects reflected impairment of acquired motivational functions, rather than with attentional processes or the display of specific learned responses. Implications of the double dissociation of these two aspects of Pavlovian conditioned incentive motivation for amygdala function in associative learning are considered. PMID- 12752387 TI - Separate processing mechanisms for encoding of geometric and landmark information in the avian hippocampus. AB - Domestic chicks bilaterally or unilaterally lesioned to the hippocampus were trained to search for food hidden beneath sawdust by ground-scratching in the centre of a large enclosure, the correct position of food being indicated by a local landmark in the absence of any extra-enclosure visual cues. At test, the landmark was removed or displaced at a distance from its original position. Results showed that sham-operated chicks and chicks with a lesion of the left hippocampus searched in the centre, relying on large-scale geometric information provided by the enclosure, whereas chicks with a lesion of either the right hippocampus or both hippocampi were completely disoriented (landmark removed) or searched close to the landmark shifted from the centre (landmark displaced). These results indicate that encoding of geometric features of an enclosure occurs in the right hippocampus even when local information provided by a landmark would suffice to localize the goal; encoding based on local information, in contrast, seems to occur outside the hippocampus. These findings provide evidence that the left and right avian hippocampi play different roles in spatial cognition, a phenomenon which had been documented previously only for the human hippocampus. PMID- 12752388 TI - Mirror neurons responding to the observation of ingestive and communicative mouth actions in the monkey ventral premotor cortex. AB - In the ventral premotor cortex (area F5) of the monkey there are neurons that discharge both when the monkey performs specific motor actions and when it observes another individual performing a similar action (mirror neurons). Previous studies on mirror neurons concerned hand actions. Here, we describe the mirror responses of F5 neurons that motorically code mouth actions. The results showed that about one-third of mouth motor neurons also discharge when the monkey observes another individual performing mouth actions. The majority of these 'mouth mirror neurons' become active during the execution and observation of mouth actions related to ingestive functions such as grasping, sucking or breaking food. Another population of mouth mirror neurons also discharges during the execution of ingestive actions, but the most effective visual stimuli in triggering them are communicative mouth gestures (e.g. lip smacking). Some also fire when the monkey makes communicative gestures. These findings extend the notion of mirror system from hand to mouth action and suggest that area F5, the area considered to be the homologue of human Broca's area, is also involved in communicative functions. PMID- 12752389 TI - Altered hemispheric asymmetry of auditory P100m in dyslexia. AB - In various studies, deviances of hemispheric laterality in the organization of the perisylvian region in dyslexia have been suggested. Although associated with impaired language functioning, the clinical significance of atypical cerebral lateralization remains unclear. The present study examined interhemispheric source differences of magnetic responses to the German synthetic syllable [ba:] in the auditory cortex of 14 dyslexic children and 12 normally literate controls aged 8-15 years. In all subjects, two main deflections, P100m and N260m, were evident in the responses over each hemisphere. While in the control group the right P100m dipole was located more anterior than the corresponding dipole of the left hemisphere, the dyslexic group displayed a rather symmetrical source configuration between the hemispheres. This symmetry reflected a deviance in the right perisylvian region for the dyslexic subjects' P100m, which was generated approximately 1 cm more posterior than the response in controls. The deviation was also obvious relative to the source location of the later component, N260m, which did not systematically differ between the participant groups. Our results suggest that the altered hemispheric asymmetry reflects an atypical organization of the right hemisphere in children and adolescents with dyslexia. PMID- 12752390 TI - Cannabinoid mechanism in reinstatement of heroin-seeking after a long period of abstinence in rats. AB - Because opioid and cannabinoid systems have been reported to interact in the modulation of addictive behaviour, this study was aimed at investigating the ability of cannabinoid agents to reinstate or prevent heroin-seeking behaviour after a prolonged period of extinction. In rats previously trained to self administer heroin intravenously, non-contingent non-reinforced priming administrations of heroin and cannabinoids were presented after long-term extinction, and lever pressing following injections was observed. Results showed that: (i) intravenous priming infusions of heroin (0.1 and 0.2 mg/kg) lead to reinstatement of drug-seeking behaviour; (ii) intraperitoneal priming injections of the central cannabinoid receptor agonists R-(+)-(2,3-dihydro-5-methyl-3-[(4 morpholinyl)methyl]pyrol[1,2,3-de]-1,4-benzoxazinyl) (1 naphthalenyl)methanonemesylate (WIN 55,212-2, 0.15 and 0.3 mg/kg) and (-)-cis-3 [2-hydroxy-4(1,1-dimethyl-heptyl)phenyl]-trans-4-(3-hydroxypropyl) cyclohexanol (CP 55,940, 0.05 and 0.1 mg/kg), but not delta9-tetrahydrocannabinol (delta9-THC, 0.1-1.0 mg/kg), effectively restored heroin-seeking behaviour; (iii) intraperitoneal priming injection of the central cannabinoid receptor antagonist N-(piperidin-1-yl)-5-(4-chlorophenyl)-1-(2,4-dichloro-phenyl)4-methyl-1H-pyrazole 3-carboxamide (SR 141716A, 0.3 mg/kg) did not reinstate responding, but (iv) completely prevented heroin-induced reinstatement of drug-seeking behaviour. Moreover, heroin-seeking behaviour was still present for a few days following cannabinoid primings, indicating a long-lasting effect of cannabinoids on responding for heroin. These findings indicate that relapse to heroin after an extended drug-free period is triggered by cannabinoid agonists and that SR 141716A prevents drug-seeking behaviour, suggesting that the use of the cannabinoid antagonist could have some therapeutic benefits in heroin-induced relapse. PMID- 12752391 TI - Acute stroke treatment in Europe: a questionnaire-based survey on behalf of the EFNS Task Force on acute neurological stroke care. AB - In 1997 the EFNS Task Force on Acute Neurological Stroke Care published its recommendations for acute stroke care (Brainin et al., 1997), which, in 1999, was followed by a stroke care inventory (Brainin et al., 2000) assessing the development of acute stroke care. In 2000 all 33 members of the EFNS Stroke Scientist Panel were asked to complete a questionnaire on the treatment of acute stroke in their country as of 2000. Data were based either on national surveys or a personal estimate of national practice, together with data from the personal practice of the panel member. Data from 22 countries were received. One of the principal findings is the lack of systematic evidence about practice in a significant number of European countries. Results illustrate that neurologists have a leading role in implementing acute stroke treatment guidelines and in adopting new therapeutic methods in most countries, but there is a wide variation in the application of different therapeutic procedures. PMID- 12752392 TI - The Education Committee of the EFNS: activities and work in progress. PMID- 12752393 TI - East-west differences in the organization of botulinum toxin use in nine Central European countries. AB - Availability and quality of expensive treatment modalities such as botulinum toxin (BTX) largely depend on organizational aspects such as costs, reimbursement by insurance companies, expertise and facilities for expert training, and the propagation of research. To investigate which determinants influence the organization of BTX' use throughout nine Central European countries (Austria, Croatia, Czech Republic, Germany, Hungary, Italy, Slovakia, Slovenia and Switzerland) we sent out questionnaires to leading BTX experts and consulted data banks of manufacturers and bulletins of international organizations. In Western European countries, there is a tendency for users to organize themselves in formal groups and to concentrate on research whereas the way how BTX is provided is diverse regarding qualifications of specialists and institutions. In the post communist Eastern European countries, we found a tendency towards a centralized system of reimbursement and BTX treatment seems to be more in the hands of neurologists than any other specialists. Strong correlations were observed between the number of BTX centres, degree of organization of user groups and number of scientific publications, on the one hand, and parameters of healthcare performance and socioeconomic determinants, on the other. Our study suggests that in the nine countries surveyed, organizational aspects of BTX use vary considerably, whilst similarities are based mainly on socioeconomic rather than socio-demographic determinants. PMID- 12752394 TI - Memory deficits and retrieval processes in ALS. AB - Subtle neuropsychological deficits have been described in patients affected by amyotrophic lateral sclerosis (ALS) without dementia. Overall, selective impairment in memory function has been reported, but the source of memory impairment in ALS has yet to be defined. We performed neuropsychological screening in 20 ALS patients. Semantic encoding and post-encoding cue effects on the retrieval of word lists were investigated in the ALS patients and normal controls. Severity of memory impairment was correlated to cerebral blood perfusion detected by single photon emission computed tomography (SPECT). ALS patients showed moderate impairments in frontal and memory tests. Short-term memory was normal, while serial position retrieval of word lists with normal recency effect but poor primacy effect showed long-term memory deficit. ALS patients performed better in cued encoding than in cued post-encoding recall condition. In the cued post-encoding condition, the primacy effect in word list recall improved significantly in controls, but not in ALS patients, as compared with both the free recall and cued encoding conditions. SPECT hypoperfusion was observed in frontal and temporal areas in ALS patients. ALS patients showed a long-term memory deficit which did not improve in cued post-encoding condition as it does for controls. We hypothesize abnormal retrieval processes related to frontal lobe dysfunction which entails difficulties in generating stable long memory traces at encoding. PMID- 12752395 TI - Sympathetic dysfunction of central origin in patients with ALS. AB - Amyotrophic lateral sclerosis (ALS) is a severe, progressive disease affecting both the central and peripheral parts of the motor nervous system. Some studies have shown unequivocal indications of a more disseminated disease also affecting the autonomic nervous system. We therefore evaluated the centrally and peripherally mediated autonomic vascular reflexes by (i) the local 133-Xenon washout technique, and (ii) the head-up tilt table test. The results correlated to clinical scores. We examined nine ALS patients and 15 age-matched controls. The 133-Xenon washout test showed a significant reduction in the centrally mediated sympathetic vasoconstrictor response, but a preserved locally mediated response in the patients. In the head-up tilt table test, the patients had a significantly higher mean arterial blood pressure (MAP) compared with controls, probably due to a general increase in vascular resistance. There were no correlations between the ALS Severity Scores and blood flow changes, diastolic blood pressure or MAP. Our study supports previous results, but indicates abnormalities consistent with a solely centrally located sympathetic dysfunction in ALS, independent of the stage of the disease. PMID- 12752396 TI - Comorbidity in post-stroke rehabilitation. AB - Patients referred to rehabilitation centers often suffer from associated comorbidity, which might negatively influence the effective outcome of the treatment program. The study was aimed at evaluating the impact of comorbidity on functional independence and gain after stroke. Ninety-three patients admitted to rehabilitation were enrolled. The disability was evaluated, both at time of admission and at discharge. The functional independence measure (FIM) was used. Comorbidity was evaluated by means of the Cumulative Illness Rating Scale (CIRS), that generates two indexes, the cumulative index (CI) and the severity index (SI). A logistic model could discriminate patients who were regularly discharged from the others (dead or transferred to acute care) pooled together (P < or = 0.02). The CI and SI were significantly correlated with FIM at admission. The r values were -0.24 (P < or = 0.02) and -0.32 (P < 0.002). Recovery was not even influenced in the most severe patients. In conclusion, the CIRS appears to be a sensitive tool for the evaluation of comorbidity in stroke patients. The comorbidity is correlated to dependence in stroke patients but does not affect functional gain. However, comorbidity is of actual interest in view of new payment systems in rehabilitation, because it is included among the variables leading to costs. PMID- 12752397 TI - Post-operative progress of dystonia patients following globus pallidus internus deep brain stimulation. AB - In the current era of functional surgery for movement disorders, deep brain stimulation (DBS) of the globus pallidus internus (GPi) is emerging as the favoured intervention for patients with dystonia. Here we report our results in 20 patients with medically intractable dystonia treated with GPi stimulation. The series comprised 14 patients with generalized dystonia and six with spasmodic torticollis. Although comparisons were limited by differences in their respective neurological rating scales, chronic DBS clearly benefited both patient groups. Data conveying the rate of change in neurological function following intervention are also presented, demonstrating the gradual but progressive and sustained nature of improvement following stimulation of the GPi in dystonic patients. PMID- 12752398 TI - Induction of prolonged tenderness in patients with tension-type headache by means of a new experimental model of myofascial pain. AB - Tenderness is the most prominent abnormal finding in patients with tension-type headache (TTH). Recently we developed a model of myofascial tenderness using intramuscular infusion of a combination of bradykinin, serotonin, histamine and prostaglandin E2. We aimed to examine tenderness after this combination in patients with episodic TTH (ETTH). Fifteen patients and 15 healthy controls completed the study. Participants received the combination into the non-dominant trapezius muscle in a randomized, double-blinded and placebo-controlled design. Local tenderness and stimulus-response functions, mechanical pain thresholds (PPDT) in the temporal region and on the finger, and total tenderness score (TTS) were recorded. A local, prolonged, and mild to moderate tenderness was reported both in patients (P = 0.001) and in controls (P = 0.001) after the combination compared with the placebo. The response to the combination tended to be increased in patients. The stimulus-response function was leftward shifted after the combination, compared with baseline in both groups. No changes in PPDT or TTS were found after the infusions, whereas baseline PPDTs were decreased in ETTH compared with controls (PPDTfinger: P = 0.033; PPDTtemporal: P = 0.015). Intramuscular infusion of a combination of endogenous substances induced prolonged tenderness in both patients with episodic TTH and healthy subjects. The present results suggest an increased excitability of peripheral muscle afferents in TTH. PMID- 12752399 TI - PET visualization of microglia in multiple sclerosis patients using [11C]PK11195. AB - Activated microglia are involved in the immune response of multiple sclerosis (MS). The peripheral benzodiazepine receptor (PBR) is expressed on microglia and up-regulated after neuronal injury. [11C]PK11195 is a positron emission tomography (PET) radioligand for the PBR. The objective of the present study was to investigate [11C]PK11195 imaging in MS patients and its additional value over magnetic resonance imaging (MRI) concerning the immuno-pathophysiological process. Seven healthy and 22 MS subjects were included. Semiquantitative [11C]PK11195 uptake values were assessed with normalization on cortical grey matter. Uptake in Gadolinium-lesions was significantly increased compared with normal white matter. Uptake in T2-lesions was generally decreased, suggesting a PBR down-regulation. However, uptake values increased whenever a clinical or MR relapse was present, suggestive for a dynamic process with a transient PBR up regulation. During disease progression, an increase of normal-appearing white matter (NAWM) uptake was found, propagating NAWM as the possible real burden of disease. In conclusion, [11C]PK11195 and PET are able to demonstrate inflammatory processes with microglial involvement in MS. PMID- 12752400 TI - Cutaneous stimulation improves function of a chronic patient with cerebellar damage. AB - The prognosis of cerebellar hemorrhage with brain stem compression is known to be poor, and patients who can usually survive are severely disabled with limited benefit from conventional rehabilitation. An innovative cutaneous stimulation was administered to a chronic patient (2 years after the incidence) who has severe ataxia, gait imbalance and limb spasticity caused by cerebellar hemorrhage. After 8 months of intervention, patient's function as evaluated by two functional measures has improved by 40%. In addition, the patient's ataxia and hypotonia have improved significantly in which he has regained the abilities to grasp objects, sit upright, control his equilibrium, and monitor an electric wheelchair. The present case study demonstrated a significant improvement of a chronic severely disabled patient who received the intervention 2 years after the accident, suggesting that the cutaneous stimulation may be a possible effective neurologic intervention. PMID- 12752401 TI - A retrospective clinical, laboratory and outcome analysis in 43 cases of acute aseptic meningitis. AB - Forty-three consecutive cases of acute aseptic meningitis (AAM) presenting within a 24-months period were retrospectively analysed with respect to clinical symptomatology, cerebrospinal fluid (CSF) findings, clinical course, treatment and outcome. Nineteen of the 43 AAM cases (44%) were caused by enterovirus, one by HIV (2%), two by Varicella zoster virus (5%), three due to herpes simplex virus I (7%), two due to herpes simplex virus II (5%), one due to Central European encephalitis virus (2%), and in 15 patients (35%) the aetiology of AAM remained unknown. Headache (100%) and fever (93%) were the presenting symptoms in the majority of cases. Signs of preceding infection were predominantly gastrointestinal in the enterovirus subgroup, but were inconsistently observed in the other subgroups. CSF findings at the first lumbar tap on admission generally revealed lymphomonocytic pleocytosis of less than 500 cells per micro l, mild to moderately elevated protein and normal lactate and glucose levels. Initial therapy consisted of an empirical antiviral and antibiotic regimen until a serological diagnosis was available. Acyclovir, effective only in herpes family viruses, was initially administered to all AAM cases. Effective therapy for other viral pathogens are not broadly available and treating AAM of unknown aetiology imposes a particular problem. The average hospitalization time ranged from 16 to 31 days. Patients were either discharged home (72%) or transferred to a rehabilitation centre (28%). The outcome was good (40%) to fair (51%) in the majority of cases. PMID- 12752402 TI - Clinical characteristics of the alpha-synuclein mutation (G209A)-associated Parkinson's disease in comparison with other forms of familial Parkinson's disease in Greece. AB - An Ala53Thr mutation of the alpha-synuclein has been recently identified as a rare cause of familial Parkinson's disease (fPD). In the present study, the clinical characteristics of Parkinson's disease (PD) patients with Ala53Thr alpha synuclein mutation (alpha-synPD) were compared with fPD patients without any known mutation. An investigator blinded to the results of the genetic analysis examined 15 alpha-synPD patients and 43 consecutive fPD patients. Demographic data, age at onset of the illness, duration of the disease and modality of presentation were collected. Segregation ratios for both sexes in individuals at risk of developing alpha-synPD were estimated. The Unified Parkinson's disease rating scale (UPDRS) was also completed. The 15 alpha-synPD patients were matched for duration of the disease and age at onset with 15 of the 43 fPD patients (MfPD). Comparisons were also made between 14 patients belonging to three multicase families with patterns of inheritance similar to alpha-synPD. The alpha synPD patients were significantly younger (mean difference 11.8 years) and showed the first sign of the disease earlier in life (mean difference 12.7 years) as compared with the fPD patients. Tremor at onset was present in only one (6.7%) of the alpha -synPD patients compared with 18 (41.9%) of the fPD patients (P = 0.01). At the time of examination rigidity, postural instability, orthostatic hypotension and the overall clinical severity did not differ significantly either between alpha-synPD and fPD or between alpha-synPD and MfPD groups. Nevertheless, some clinically relevant trends concerning the psychiatric symptoms and complications of therapy were recognized. The overall clinical severity and the progression of the disease in patients with alpha-synPD did not differ from that of the fPD patients. The alpha-synPD patients presented the illness at a younger age and also had lower prevalence of tremor when compared with the fPD patients. PMID- 12752403 TI - Expectations of wheelchair-dependency in recently diagnosed patients with multiple sclerosis and their partners. AB - The aim of the present paper was to quantify expectations of wheelchair dependency in patients recently diagnosed with MS (n = 101) and their partners (n = 78). Expectations focused on the risk and seriousness of becoming wheelchair dependent in 2 years, 10 years or lifetime. Expectations were compared with natural history data, compared between patients and their partners, and related to clinical characteristics. Our results show that patients overestimated their 2 year and 10-year risks of wheelchair-dependency, but underestimated their lifetime risks. A large number of patients were uncertain about their 2-year risk, even those with no or only minimal disability [Expanded Disability Status Scale (EDSS) <3.0]. One-third of the patients perceived the 10-year and lifetime risk to be 50%, which, as they explained in the interviews, reflected their uncertainty: they did not know what to expect - it might happen or not. Patients with more functional limitations had higher perceptions of risk, but lower perceptions of seriousness. Concordance in perceived risk and seriousness between patients and partners was moderate. The overestimation of the short-term risks and the substantial differences in expectations within couples warrant further research on the impact of expectations on their treatment decisions and psychological well-being. PMID- 12752404 TI - Nociceptive-specific blink reflex and glyceryl trinitrate infusion in healthy volunteers. AB - Glyceryl trinitrate (GTN) is known to induce early headache in healthy humans after intravenous infusion. Moreover, in animal models subcutaneous administration produces an increase in Fos expression in brainstem areas that are involved in trigeminal pain processing. In a double-blind crossover study, we tested the blink reflex before, during and immediately after GTN and placebo intravenous infusion in eight healthy volunteers using a new stimulation electrode that preferentially activates A-delta nociceptive afferent fibres. The initial hypothesis that GTN could induce an increase in the magnitude of the nociceptive blink reflex R2 component by stimulating activity of trigeminal nucleus caudalis wide dynamic range interneurones was not confirmed. Although mild headache was induced in six subjects, there was no significant change between the R2 area under the curve before and after drug vs. placebo. PMID- 12752405 TI - Quality of neurology residency programmes: an Italian survey. AB - In recent years there has been a growing interest in medical and particularly neurological education and how this should be related to the needs for patient care. To evaluate neurological training in Italy, we conducted a survey of the residency programmes aimed at different aspects of training. The survey was conducted in the 38 neurological Italian teaching hospitals and 27 of these answered. Six of the 27 centres organized all of the scheduled teaching courses. The quality of courses was considered 'not sufficient' in 11 schools and 'good' in 12. Seminars were regularly performed in 18 centres but in 60% of these the number was <1 per week. Questionnaires to evaluate the quality of teaching were lacking in all centres. Regarding the procedures performed by each resident there was a large variation between the different schools. A regular rotation of each resident in the neurophysiology services was performed in 14 schools. Ward and out patient activity varied widely and details are given. We conclude that there is marked heterogeneity in training programmes between different centres. Some important activities such as seminars and rotation in neurophysiology are performed poorly. PMID- 12752406 TI - The trigemino-cervical reflex in tension-type headache. AB - To investigate the pathophysiology of tension-type headache (TTH) with special reference to central mechanisms and to the involvement of the trigeminal system. Short latency responses can be recorded in tonically active sternocleidomastoid muscle after stimulation of the infraorbital branch of the trigeminal nerve (the trigemino-cervical reflex). This brainstem reflex was studied in 15 healthy subjects, in 15 patients with episodic tension-type headache (ETTH) and in 15 patients with chronic tension-type headache (CTTH) outside of the pain attacks. The trigemino-cervical response was abnormal, in the size or latency, in 13 patients with CTTH and in only one patient with ETTH. This finding strongly suggests that only in the CTTH the underlying pathophysiology involves the trigeminal system. The trigemino-cervical reflex is a sensitive method to evaluate the involvement of the trigeminal brainstem neurones in TTH and their assessment may provide useful diagnostic and prognostic information. PMID- 12752407 TI - Functional improvement in cerebral palsy patients treated with botulinum toxin A injections - preliminary results. AB - Authors report the preliminary results of an open-label, prospective study to evaluate a functional benefit of botulinum toxin type A injections in diparetic cerebral palsy patients, using gross motor function measure (GMFM) score. In a group of 14 children (mean age 3.9 years, range 2-6) treated with Dysport 30 IU/kg, a statistically significant improvement (P < 0.05) was noticed in both simple measurements (Modified Ashworth Scale, Selective Motor Control, Passive Range of Movements, Physician Rating Scale and parental Clinical Global Impression) and complex functions (GMFM dimensions D and E) after 1 and 3 months. However, the simple measurement scores decreased (but not to the baseline) after 3 months; surprisingly, GMFM scores were still increasing (7.7% change after 3 months and 11.3% change after 6 months in nine patients). These results are in concordance with a few other data published to date. The study may support the concept of persistent functional gain in long-term treatment of spasticity caused by cerebral palsy with botulinum toxin type A. PMID- 12752408 TI - Early-onset Alzheimer's disease with presenilin-1 M139V mutation: clinical, neuropsychological and neuropathological study. AB - The clinical, neuropsychological and neuropathological features of a patient with early-onset Alzheimer's disease as a result of the M139V presenilin-1 (PSEN-1) mutation are presented, and compared with previous reports of patients with the same mutation. Similarities, such as the age at onset and the relative preservation of naming skills, and differences, such as the significant basal ganglia, thalamic and cerebellar pathology, are noted. This clinical and pathological heterogeneity in patients with the same PSEN-1 mutation suggests phenotype modulation by genetic and/or epigenetic factors. PMID- 12752409 TI - Extensive MRI lesion in brain stem of a neuro-Behcet patient. PMID- 12752410 TI - Writer's cramp in a surgeon. PMID- 12752411 TI - Central nervous system demyelination in familial Mediterranean fever: is it a coincidence? PMID- 12752412 TI - Lhermitte's sign following oxaliplatin-containing chemotherapy in a cisplatin pretreated ovarian cancer patient. PMID- 12752413 TI - MRI findings in acute hypertensive encephalopathy. PMID- 12752414 TI - Bilateral meralgia paraesthetica following repeated laparotomies. PMID- 12752415 TI - Anticoagulation of an unruptured, thrombosed giant intracranial aneurysm without hemorrhage or recanalization in the long-term follow-up. PMID- 12752416 TI - Cerebral infarction in patient with colon carcinoma and protein C and S deficiencies. PMID- 12752417 TI - Hippocampal sclerosis in a case of Alzheimer's disease-like dementia with late onset intractable epilepsy. PMID- 12752431 TI - Characterization of surface n-alkanes and fatty acids of the epiphytic lichen Xanthoria parietina, its photobiont a green alga Trebouxia sp., and its mycobiont, from the Jerusalem hills. AB - Surface alkanes and fatty acids from the thalli of the lichen Xanthoria parietina, its photobiont Trebouxia sp., and its mycobiont were analysed by GC MS. The green alga Trebouxia sp. synthesized mainly unsaturated fatty acids such as (Z,Z,Z)-9,12,15-18 : 3 (Z,Z)-9,12-18 : 2 and (Z)-9-18 : 1, and light alkanes C8-C15 (up to 83% of total n-alkanes). However, the mycobiont contained mainly saturated fatty acids such as hexadecanoic (16 : 0) and octadecanoic acid (18 : 0), and also very long-chain n-alkanes C22-C34. Dehydroabietic acid was found in both lichen and mycobiont. The occurrence of different amounts of n-alkanes and fatty acids in the photobionts and mycobionts of X. parietina was shown for the first time. Lichens collected from different locations in the Jerusalem hills contained n-alkanes ranging in concentration from 187 to 211 mg x (g dry wt)-1; n alkane concentrations in the photobiont and mycobiont were 17-24 and 215-262 mg x (g dry wt)-1, respectively. PMID- 12752430 TI - The mystery of nonclassical protein secretion. A current view on cargo proteins and potential export routes. AB - Most of the examples of protein translocation across a membrane (such as the import of classical secretory proteins into the endoplasmic reticulum, import of proteins into mitochondria and peroxisomes, as well as protein import into and export from the nucleus), are understood in great detail. In striking contrast, the phenomenon of unconventional protein secretion (also known as nonclassical protein export or ER/Golgi-independent protein secretion) from eukaryotic cells was discovered more than 10 years ago and yet the molecular mechanism and the molecular identity of machinery components that mediate this process remain elusive. This problem appears to be even more complex as several lines of evidence indicate that various kinds of mechanistically distinct nonclassical export routes may exist. In most cases these secretory mechanisms are gated in a tightly controlled fashion. This review aims to provide a comprehensive overview of our current knowledge as a basis for the development of new experimental strategies designed to unravel the molecular machineries mediating ER/Golgi independent protein secretion. Beyond solving a fundamental problem in current cell biology, the molecular analysis of these processes is of major biomedical importance as these export routes are taken by proteins such as angiogenic growth factors, inflammatory cytokines, components of the extracellular matrix which regulate cell differentiation, proliferation and apoptosis, viral proteins, and parasite surface proteins potentially involved in host infection. PMID- 12752432 TI - Tracking interactions that stabilize the dimer structure of starch phosphorylase from Corynebacterium callunae. Roles of Arg234 and Arg242 revealed by sequence analysis and site-directed mutagenesis. AB - Glycogen phosphorylases (GPs) constitute a family of widely spread catabolic alpha1,4-glucosyltransferases that are active as dimers of two identical, pyridoxal 5'-phosphate-containing subunits. In GP from Corynebacterium callunae, physiological concentrations of phosphate are required to inhibit dissociation of protomers and cause a 100-fold increase in kinetic stability of the functional quarternary structure. To examine interactions involved in this large stabilization, we have cloned and sequenced the coding gene and have expressed fully active C. callunae GP in Escherichia coli. By comparing multiple sequence alignment to structure-function assignments for regulated and nonregulated GPs that are stable in the absence of phosphate, we have scrutinized the primary structure of C. callunae enzyme for sequence changes possibly related to phosphate-dependent dimer stability. Location of Arg234, Arg236, and Arg242 within the predicted subunit-to-subunit contact region made these residues primary candidates for site-directed mutagenesis. Individual Arg-->Ala mutants were purified and characterized using time-dependent denaturation assays in urea and at 45 degrees C. R234A and R242A are enzymatically active dimers and in the absence of added phosphate, they display a sixfold and fourfold greater kinetic stability of quarternary interactions than the wild-type, respectively. The stabilization by 10 mm of phosphate was, however, up to 20-fold greater in the wild-type than in the two mutants. The replacement of Arg236 by Ala was functionally silent under all conditions tested. Arg234 and Arg242 thus partially destabilize the C. callunae GP dimer structure, and phosphate binding causes a change of their tertiary or quartenary contacts, likely by an allosteric mechanism, which contributes to a reduced protomer dissociation rate. PMID- 12752433 TI - Dissociation of DNA polymerase alpha-primase complex during meiosis in Coprinus cinereus. AB - Previously, the activity of DNA polymerase alpha was found in the meiotic prophase I including non-S phase stages, in the basidiomycetes, Coprinus cinereus. To study DNA polymerase alpha during meiosis, we cloned cDNAs for the C. cinereus DNA polymerase alpha catalytic subunit (p140) and C. cinereus primase small subunit (p48). Northern analysis indicated that both p140 and p48 are expressed not only at S phase but also during the leptotene/zygotene stages of meiotic prophase I. In situ immuno-staining of cells at meiotic prophase I revealed a sub population of p48 that does not colocalize with p140 in nuclei. We also purified the pol alpha-primase complex from meiotic cells by column chromatography and characterized its biochemical properties. We found a subpopulation of primase that was separated from the pol alpha-primase complex by phosphocellulose column chromatography. Glycerol gradient density sedimentation results indicated that the amount of intact pol alpha-primase complex in crude extract is reduced, and that a smaller complex appears upon meiotic development. These results suggest that the form of the DNA polymerase alpha-primase complex is altered during meiotic development. PMID- 12752435 TI - A unique variant of streptococcal group O-antigen (C-polysaccharide) that lacks phosphocholine. AB - Streptococcus mitis strain SK598, which represents a subgroup of biovar 1, possesses a unique variant of the C-polysaccharide found in the cell wall of all strains of Streptococcus pneumoniae and in some strains of S. mitis. This new variant lacks the choline methyl groups in contrast to the previously characterized forms of C-polysaccharide, which all contain one or two choline residues per repeat. The following structure of the repeating unit of the SK598 polysaccharide was established: where AAT is 2-acetamido-4-amino-2,4,6-trideoxy-d galactose. This structure is identical to the double choline-substituted form of C-polysaccharide, except that it is substituted with ethanolamine instead of choline. This extends the number of recognized C-polysaccharide variants to four. PMID- 12752434 TI - Different modes of dipeptidyl peptidase IV (CD26) inhibition by oligopeptides derived from the N-terminus of HIV-1 Tat indicate at least two inhibitor binding sites. AB - Dipeptidyl peptidase IV (DP IV, CD26) plays an essential role in the activation and proliferation of lymphocytes, which is shown by the immunosuppressive effects of synthetic DP IV inhibitors. Similarly, both human immunodeficiency virus-1 (HIV-1) Tat protein and the N-terminal peptide Tat(1-9) inhibit DP IV activity and T cell proliferation. Therefore, the N-terminal amino acid sequence of HIV-1 Tat is important for the inhibition of DP IV. Recently, we characterized the thromboxane A2 receptor peptide TXA2-R(1-9), bearing the N-terminal MWP sequence motif, as a potent DP IV inhibitor possibly playing a functional role during antigen presentation by inhibiting T cell-expressed DP IV [Wrenger, S., Faust, J., Mrestani-Klaus, C., Fengler, A., Stockel-Maschek, A., Lorey, S., Kahne, T., Brandt, W., Neubert, K., Ansorge, S. & Reinhold, D. (2000) J. Biol. Chem.275, 22180-22186]. Here, we demonstrate that amino acid substitutions at different positions of Tat(1-9) can result in a change of the inhibition type. Certain Tat(1-9)-related peptides are found to be competitive, and others linear mixed type or parabolic mixed-type inhibitors indicating different inhibitor binding sites on DP IV, at the active site and out of the active site. The parabolic mixed-type mechanism, attributed to both non-mutually exclusive inhibitor binding sites of the enzyme, is described in detail. From the kinetic investigations and molecular modeling experiments, possible interactions of the oligopeptides with specified amino acids of DP IV are suggested. These findings give new insights for the development of more potent and specific peptide-based DP IV inhibitors. Such inhibitors could be useful for the treatment of autoimmune and inflammatory diseases. PMID- 12752436 TI - Comparison of the solution structures of angiotensin I & II. Implication for structure-function relationship. AB - Conformational analysis of angiotensin I (AI) and II (AII) peptides has been performed through 2D 1H-NMR spectroscopy in dimethylsulfoxide and 2,2,2 trifluoroethanol/H2O. The solution structural models of AI and AII have been determined in dimethylsulfoxide using NOE distance and 3JHNHalpha coupling constants. Finally, the AI family of models resulting from restrained energy minimization (REM) refinement, exhibits pairwise rmsd values for the family ensemble 0.26 +/- 0.13 A, 1.05 +/- 0.23 A, for backbone and heavy atoms, respectively, and the distance penalty function is calculated at 0.075 +/- 0.006 A2. Comparable results have been afforded for AII ensemble (rmsd values 0.30 +/- 0.22 A, 1.38 +/- 0.48 A for backbone and heavy atoms, respectively; distance penalty function is 0.029 +/- 0.003 A2). The two peptides demonstrate similar N terminal and different C-terminal conformation as a consequence of the presence/absence of the His9-Leu10 dipeptide, which plays an important role in the different biological function of the two peptides. Other conformational variations focused on the side-chain orientation of aromatic residues, which constitute a biologically relevant hydrophobic core and whose inter-residue contacts are strong in dimethylsulfoxide and are retained even in mixed organic aqueous media. Detailed analysis of the peptide structural features attempts to elucidate the conformational role of the C-terminal dipeptide to the different binding affinity of AI and AII towards the AT1 receptor and sets the basis for understanding the factors that might govern free- or bound-depended AII structural differentiation. PMID- 12752437 TI - Phosphorylation of NF-kappaB proteins by cyclic GMP-dependent kinase. A noncanonical pathway to NF-kappaB activation. AB - The transcription factor NF-kappaB is activated in cellular stress responses. This requires rapid regulation of its function, which is accomplished, in part, by various modes of phosphorylation. Even though diverse DNA binding subunits of NF-kappaB proteins may transactivate from distinct recognition sequences, the differential regulation of transcription from the large number of NF-kappaB responsive sites in various gene promoters and enhancers has been incompletely understood. The cyclic GMP-dependent kinase (PKG) is an important mediator of signal transduction that may induce gene expression through cAMP response element binding protein (CREB) and through other, yet undefined, mechanisms. We have previously characterized a signal transduction pathway that leads to activation induced cell death in T-lymphocytes and involves the activation of PKG. Here we demonstrate that the NF-kappaB proteins p65, p49 (also called p52), and p50 are specific substrates for this kinase. PKG dose-dependently increases the transactivating activity of p65 from the NF-kappaB consensus sequence. It also mediates dose-dependently an increase in transcriptional activity by p49 or p50 from a unique CCAAT/enhance binding protein (C/EBP)-associated NF-kappaB site, but not from the consensus site. Phosphorylation of p65, p50, or p49 does not alter their subcellular distribution. Because the release of cytosolic p65/p50 heterodimers into the nucleus is by itself insufficient to differentiate all the numerous NF-kappaB promoter sequences, phosphorylation of the DNA-binding subunits reveals a form of differential regulation of NF-kappaB activity and it implies a novel pathway for PKG-induced gene transcription. These observations may bear on mechanisms of programmed cell death in T-lymphocytes. They may also be relevant to ongoing efforts to induce cancer cell apoptosis through activation of PKG. PMID- 12752438 TI - An alternative mechanism of product chain-length determination in type III geranylgeranyl diphosphate synthase. AB - (All-E) prenyl diphosphate synthases catalyze the consecutive condensation of isopentenyl diphosphates with allylic prenyl diphosphates, producing products with various chain-lengths that are unique for each enzyme. Some short-chain (all E) prenyl diphosphate synthases, i.e. farnesyl diphosphate synthases and geranylgeranyl diphosphate synthases contain characteristic amino acid sequences around the allylic substrate binding sites, which have been shown to play a role in determining the chain-length of the product. However, among these enzymes, which are classified into several types based on the possessive patterns of such characteristics, type III geranylgeranyl diphosphate synthases, which consist of enzymes from eukaryotes (excepting plants), lack these features. In this study, we report that mutagenesis at the second position before the conserved G(Q/E) motif, which is distant from the well-studied region, affects the chain-length of the product for a type III geranylgeranyl diphosphate synthase from Saccharomyces cerevisiae. This clearly suggests that a novel mechanism is operative in the product determination for this type of enzyme. We also show herein that mutagenesis at the corresponding position of an archaeal medium-chain enzyme also alters its product specificity. These results provide valuable information on the molecular evolution of (all-E) prenyl diphosphate synthases. PMID- 12752439 TI - Limited proteolysis of Escherichia coli cytidine 5'-triphosphate synthase. Identification of residues required for CTP formation and GTP-dependent activation of glutamine hydrolysis. AB - Cytidine 5'-triphosphate synthase catalyses the ATP-dependent formation of CTP from UTP using either ammonia or l-glutamine as the source of nitrogen. When glutamine is the substrate, GTP is required as an allosteric effector to promote catalysis. Limited trypsin-catalysed proteolysis, Edman degradation, and site directed mutagenesis were used to identify peptide bonds C-terminal to three basic residues (Lys187, Arg429, and Lys432) of Escherichia coli CTP synthase that were highly susceptible to proteolysis. Lys187 is located at the CTP/UTP-binding site within the synthase domain, and cleavage at this site destroyed all synthase activity. Nucleotides protected the enzyme against proteolysis at Lys187 (CTP > ATP > UTP > GTP). The K187A mutant was resistant to proteolysis at this site, could not catalyse CTP formation, and exhibited low glutaminase activity that was enhanced slightly by GTP. K187A was able to form tetramers in the presence of UTP and ATP. Arg429 and Lys432 appear to reside in an exposed loop in the glutamine amide transfer (GAT) domain. Trypsin-catalyzed proteolysis occurred at Arg429 and Lys432 with a ratio of 2.6 : 1, and nucleotides did not protect these sites from cleavage. The R429A and R429A/K432A mutants exhibited reduced rates of trypsin catalyzed proteolysis in the GAT domain and wild-type ability to catalyse NH3 dependent CTP formation. For these mutants, the values of kcat/Km and kcat for glutamine-dependent CTP formation were reduced approximately 20-fold and approximately 10-fold, respectively, relative to wild-type enzyme; however, the value of Km for glutamine was not significantly altered. Activation of the glutaminase activity of R429A by GTP was reduced 6-fold at saturating concentrations of GTP and the GTP binding affinity was reduced 10-fold. This suggests that Arg429 plays a role in both GTP-dependent activation and GTP binding. PMID- 12752440 TI - The antibiotic activity of cationic linear amphipathic peptides: lessons from the action of leucine/lysine copolymers on bacteria of the class Mollicutes. AB - Peptides composed of leucyl and lysyl residues ('LK peptides') with different compositions and sequences were compared for their antibacterial activities using cell wall-less bacteria of the class Mollicutes (acholeplasmas, mycoplasmas and spiroplasmas) as targets. The antibacterial activity of the amphipathic alpha helical peptides varied with their size, 15 residues being the optimal length, independent of the membrane hydrophobic core thickness and the amount of cholesterol. The 15-residue ideally amphipathic alpha helix with a +5 positive net charge (KLLKLLLKLLLKLLK) had the strongest antibacterial activity, similar to that of melittin. In contrast, scrambled peptides devoid of amphipathy and the less hydrophobic beta-sheeted peptides [(LK)nK], even those 15-residue long, were far less potent than the helical ones. Furthermore, the growth inhibitory activity of the peptides was correlated with their ability to abolish membrane potential. These data are fully consistent with a predominantly flat orientation of LK peptides at the lipid/water interface and strongly supports that these peptides and probably the linear polycationic amphipathic defence peptides act on bacterial membranes in four main steps according to the 'carpet' model: (a) interfacial partitioning with accumulation of monomers on the target membrane (limiting step); (b) peptide structural changes (conformation, aggregation, and orientation) induced by interactions with the lipid bilayer (as already shown with liposomes and erythrocytes); (c) plasma membrane permeabilization/depolarization via a detergent-like effect; and (d) rapid bacterial cell death if the extent of depolarization is maintained above a critical threshold. PMID- 12752441 TI - Accessory proteins functioning selectively and pleiotropically in the biosynthesis of [NiFe] hydrogenases in Thiocapsa roseopersicina. AB - There are at least two membrane-bound (HynSL and HupSL) and one soluble (HoxEFUYH) [NiFe] hydrogenases in Thiocapsa roseopersicina BBS, a purple sulfur photosynthetic bacterium. Genes coding for accessory proteins that participate in the biosynthesis and maturation of hydrogenases seem to be scattered along the chromosome. Transposon-based mutagenesis was used to locate the hydrogenase accessory genes. Molecular analysis of strains showing mutant phenotypes led to the identification of hupK (hoxV ), hypC1, hypC2, hypD, hypE, and hynD genes. The roles of hynD, hupK and the two hypC genes were investigated in detail. The putative HynD was found to be a hydrogenase-specific endoprotease type protein, participating in the maturation of the HynSL enzyme. HupK plays an important role in the formation of the functionally active membrane-bound [NiFe] hydrogenases, but not in the biosynthesis of the soluble enzyme. In-frame deletion mutagenesis showed that HypC proteins were not specific for the maturation of either hydrogenase enzyme. The lack of either HypC protein drastically reduced the activity of every hydrogenase. Hence both HypCs might participate in the maturation of [NiFe] hydrogenases. Homologous complementation with the appropriate genes substantiated the physiological roles of the corresponding gene products in the H2 metabolism of T. roseopersicina. PMID- 12752442 TI - The cellular oxygen tension regulates expression of the endoplasmic oxidoreductase ERO1-Lalpha. AB - The formation of disulfide bonds in the endoplasmic reticulum requires protein disulfide isomerase (PDI) and endoplasmic reticulum oxidoreductin 1 (ERO1) that reoxidizes PDI. We report here that the expression of the rat, mouse and human homologues of ERO1-Like protein alpha but not of the isoform ERO1-Lbeta are stimulated by hypoxia in rats vivo and in rat, mouse and human cell cultures. The temporal pattern of hypoxic ERO1-Lalpha induction is very similar to that of genes triggered by the hypoxia inducible transcription factor (HIF-1) and is characteristically mimicked by cobalt and by deferoxamine, but is absent in cells with a defective aryl hydrocarbon receptor translocator (ARNT, HIF-1beta). We speculate from these findings that the expression of ERO1-Lalpha is probably regulated via the HIF-pathway and thus belongs to the family of classic oxygen regulated genes. Activation of the unfolded protein response (UPR) by tunicamycin, on the other hand, strongly induced ERO1-Lbeta and more moderately ERO1-Lalpha expression. The expression of the two ERO1-L isoforms therefore appears to be differently regulated, in the way that ERO1-Lalpha expression is mainly controlled by the cellular oxygen tension, whilst ERO1-Lbeta is triggered mainly by UPR. The physiological meaning of the oxygen regulation of ERO1-Lalpha expression likely is to maintain the transfer rate of oxidizing equivalents to PDI in situations of an altered cellular redox state induced by changes of the cellular oxygen tension. PMID- 12752443 TI - Co-operation of domain-binding and calcium-binding sites in the activation of gelsolin. AB - Gelsolin is an abundant calcium dependent actin filament severing and capping protein. In the absence of calcium the molecule is compact but in the presence of calcium, as its six similar domains alter their relative position, a generally more open configuration is adopted to reveal the three actin binding sites. It is generally held that a 'helical-latch' at the C-terminus of gelsolin's domain 6 (G6), binds domain 2 (G2) to keep gelsolin in the calcium-free compact state, and that the crutial calcium binding site(s) reside in the C-terminal half of gelsolin perhaps involving the C-terminal helix itself has to be bound to release this latch. Here we provide evidence for a calcium dependent conformational change within G2 (Kd = approximately 15 micro m). We also report a calcium dependent binding site for the C-terminus (G4-6) within G2 and delimit this further to a specific region formed by residues 203-225 and 159-193. It is known that the activation of gelsolin involves multiple calcium binding events (around 6) the first of which (in G6) may release the latch. We propose that the calcium dependent conformational change in G2 may be a subsequent step that is necessary for the dissociation of G2 from G4-6, and that this movement occurs in sympathy with calcium induced conformational changes within G6 by the physical coupling of the two calcium binding sites within G2 and G6. Additional calcium binding in other domains then result in the complete opening and activation of the gelsolin molecule. PMID- 12752444 TI - The oxygenase component of phenol hydroxylase from Acinetobacter radioresistens S13. AB - Phenol hydroxylase (PH) from Acinetobacter radioresistens S13 represents an example of multicomponent aromatic ring monooxygenase made up of three moieties: a reductase (PHR), an oxygenase (PHO) and a regulative component (PHI). The function of the oxygenase component (PHO), here characterized for the first time, is to bind molecular oxygen and catalyse the mono-hydroxylation of substrates (phenol, and with less efficiency, chloro- and methyl-phenol and naphthol). PHO was purified from extracts of A. radioresistens S13 cells and shown to be a dimer of 206 kDa. Each monomer is composed by three subunits: alpha (54 kDa), beta (38 kDa) and gamma (11 kDa). The gene encoding PHO alpha (named mopN) was cloned and sequenced and the corresponding amino acid sequence matched with that of functionally related oxygenases. By structural alignment with the catalytic subunits of methane monooxygenase (MMO) and alkene monooxygenase, we propose that PHO alpha contains the enzyme active site, harbouring a dinuclear iron centre Fe O-Fe, as also suggested by spectral analysis. Conserved hydrophobic amino acids known to define the substrate recognition pocket, are also present in the alpha subunit. The prevalence of alpha-helices (99.6%) as studied by CD confirmed the hypothized structural homologies between PHO and MMO. Three parameters (optimum ionic strength, temperature and pH) that affect kinetics of the overall phenol hydroxylase reaction were further analyzed with a fixed optimal PHR/PHI/PHO ratio of 2/1/1. The highest level of activity was evaluated between 0.075 and 0.1 m of ionic strength, the temperature dependence showed a maximum of activity at 24 degrees C and finally the pH for optimal activity was determined to be 7.5. PMID- 12752445 TI - The histidine-phosphocarrier protein of Streptomyces coelicolor folds by a partially folded species at low pH. AB - The folding of a 93-residue protein, the histidine-phosphocarrier protein of Streptomyces coelicolor, HPr, has been studied using several biophysical techniques, namely fluorescence, 8-anilinonaphthalene-1-sulfate binding, circular dichroism, Fourier transform infrared spectroscopy, gel filtration chromatography and differential scanning calorimetry. The chemical-denaturation behaviour of HPr, followed by fluorescence, CD and gel filtration, at pH 7.5 and 25 degrees C, is described as a two-state process, which does not involve the accumulation of thermodynamically stable intermediates. Its conformational stability under those conditions is deltaG = 4.0 +/- 0.2 kcal x mol-1 (1 kcal = 4.18 kJ), which makes the HPr from S. coelicolor the most unstable member of the HPr family described so far. The stability of the protein does not change significantly from pH 7-9, as concluded from the differential scanning calorimetry and thermal CD experiments. Conformational studies at low pH (pH 2.5-4) suggest that, in the absence of cosmotropic agents, HPr does not unfold completely; rather, it accumulates partially folded species. The transition from those species to other states with native-like secondary and tertiary structure, occurs with a pKa = 3.3 +/- 0.3, as measured by the averaged measurements obtained by CD and fluorescence. However, this transition does not agree either with: (a) that measured by burial of hydrophobic patches (8-anilinonaphthalene-1-sulfate binding experiments); or (b) that measured by acquisition of native-like compactness (gel filtration studies). It seems that acquisition of native-like features occurs in a wide pH range and it cannot be ascribed to a unique side-chain titration. These series of intermediates have not been reported previously in any member of the HPr family. PMID- 12752446 TI - Cytochrome b559 content in isolated photosystem II reaction center preparations. AB - The cytochrome b559 content was examined in five types of isolated photosystem II D1-D2-cytochrome b559 reaction center preparations containing either five or six chlorophylls per reaction center. The reaction center complexes were obtained following isolation procedures that differed in chromatographic column material, washing buffer composition and detergent concentration. Two different types of cytochrome b559 assays were performed. The absolute heme content in each preparation was obtained using the oxidized-minus-reduced difference extinction coefficient of cytochrome b559 at 559 nm. The relative amount of D1 and cytochrome b559alpha-subunit polypeptide was also calculated for each preparation from immunoblots obtained using antibodies raised against the two polypeptides. The results indicate that the cytochrome b559 heme content in photosystem II reaction center complexes can vary with the isolation procedure, but the variation of the cytochrome b559alpha-subunit/D1 polypeptide ratio was even greater. This variation was not found in the PSII-enriched membrane fragments used as the RC-isolation starting material, as different batches of membranes obtained from spinach harvested at different seasons of the year or those from sugar beets grown in a chamber under controlled environmental conditions lack variation in their alpha-subunit/D1 polypeptide ratio. A precise determination of the ratio using an RC1-control sample calibration curve gave a ratio of 1.25 cytochrome b559alpha-subunit per 1.0 D1 polypeptide in photosystem II membranes. We conclude that the variations found in the reaction center preparations were due to the different procedures used to isolate and purify the different reaction center complexes. PMID- 12752447 TI - Gene transcription of fgl2 in endothelial cells is controlled by Ets-1 and Oct-1 and requires the presence of both Sp1 and Sp3. AB - The immune coagulant fgl2/fibroleukin has been previously shown to play a pivotal role in the pathogenesis of murine and human fulminant hepatitis and fetal loss syndrome. Constitutive expression of fgl2 transcripts at low levels are seen in cytotoxic T cells, endothelial, intestinal and trophoblast cells, while specific factors (such as virus and cytokines) are required to induce high levels of fgl2 expression in other cell types including monocytes/macrophages. To address the transcriptional mechanisms that regulate constitutive expression of fgl2, murine genomic clones were characterized and the transcription start site was defined by 5'-RACE and primer extension. A comprehensive assessment of basal fgl2 promoter activity in murine vascular endothelial cells defined a minimal 119 bp region responsible for constitutive fgl2 transcription. A complex positive regulatory domain (PRD) spanning a 39-bp sequence from -87 to -49 (relative to the transcription start site) was identified. Electrophoretic mobility shift assay studies in vascular endothelial cells revealed that the nucleoprotein complexes that form on this positive regulatory domain (PRD) contain Sp1/Sp3 family members, Oct-1, and Ets-1. Heterologous expression studies in Drosophila Schneider cells confirmed that the constitutive expression of this gene is controlled by Ets-1 and requires the presence both of the Sp1 and Sp3 transcription factors. The presence of this complex multicomponent PRD in the fgl2 proximal promoter is consistent with the observation that, in vivo, fgl2 expression is tightly regulated. Moreover, viral induced fgl2 expression also requires the presence of this PRD. These results clearly demonstrate that multiple cis DNA elements in a clustered region work cooperatively to regulate constitutive fgl2 expression and interact with inducible elements to regulate viral-induced fgl2 expression in endothelial cells. PMID- 12752448 TI - A peptide containing a novel FPGN CD40-binding sequence enhances adenoviral infection of murine and human dendritic cells. AB - CD40 is a receptor with numerous functions in the activation of antigen presenting cells (APCs), particularly dendritic cells (DC). Using phage display technology, we identified linear peptides containing a novel FPGN/S consensus sequence that enhances the binding of phage to a purified murine CD40 immunoglobulin (Ig) fusion protein (CD40-Ig), but not to Ig alone. To examine the ability the FPGN/S peptides to enhance adenoviral infection of CD40-positive cells, we used bifunctional peptides consisting of an FPGN-containing peptide covalently linked to an adenoviral knob-binding peptide (KBP). One of these, FPGN2-KBP, was able to enhance adenoviral infection of both murine and human DCs in a dose-dependent manner. FPGN2-KBP also improved infection of murine B cell blasts, a murine B lymphoma cell line (L10A), and immortalized human B cells. To demonstrate that enhancement of adenoviral infection depended on the presence of CD40, we analyzed infection of the breast cancer line, SKBR3, that does not express CD40 or the adenovirus cellular receptor, CAR. Infection of SKBR3 cells was enhanced by FPGN2-KBP following transient transfection with a plasmid vector that expresses murine CD40, but not when the cells were mock-transfected. In conclusion, we have isolated a peptide that binds to murine CD40, and promotes the uptake of adenoviruses into CD40-expressing cells of both murine and human origin, suggesting that it may have potential applications for antigen delivery to CD40-positive antigen-presenting cells. PMID- 12752449 TI - Changes in rat liver mitochondria with aging. Lon protease-like reactivity and N(epsilon)-carboxymethyllysine accumulation in the matrix. AB - Aging is accompanied by a gradual deterioration of cell functions. Mitochondrial dysfunction and accumulation of protein damage have been proposed to contribute to this process. The present study was carried out to examine the effects of aging in mitochondrial matrix isolated from rat liver. The activity of Lon protease, an enzyme implicated in the degradation of abnormal matrix proteins, was measured and the accumulation of oxidation and glycoxidation (Nepsilon carboxymethyllysine, CML) products was monitored using immunochemical assays. The function of isolated mitochondria was assessed by measuring respiratory chain activity. Mitochondria from aged (27 months) rats exhibited the same rate of oxygen consumption as those from adult (10 months) rats without any change in coupling efficiency. At the same time, the ATP-stimulated Lon protease activity, measured as fluorescent peptides released, markedly decreased from 10-month-old rats (1.15 +/- 0.15 FU x micro g protein-1 x h-1) to 27-month-old-rats (0.59 +/- 0.08 FU x micro g protein-1 x h-1). In parallel with this decrease in activity, oxidized proteins accumulated in the matrix upon aging while the CML-modified protein content assessed by ELISA significantly increased by 52% from 10 months (11.71 +/- 0.61 pmol CML x micro g protein-1) to 27 months (17.81 +/- 1.83 pmol CML x micro g protein-1). These results indicate that the accumulation of deleterious oxidized and carboxymethylated proteins in the matrix concomitant with loss of the Lon protease activity may affect the ability of aging mitochondria to respond to additional stress. PMID- 12752450 TI - Interactions between M proteins of Streptococcus pyogenes and glycosaminoglycans promote bacterial adhesion to host cells. AB - Several microbial pathogens have been reported to interact with glycosaminoglycans (GAGs) on cell surfaces and in the extracellular matrix. Here we demonstrate that M protein, a major surface-expressed virulence factor of the human bacterial pathogen, Streptococcus pyogenes, mediates binding to various forms of GAGs. Hence, S. pyogenes strains expressing a large number of different types of M proteins bound to dermatan sulfate (DS), highly sulfated fractions of heparan sulfate (HS) and heparin, whereas strains deficient in M protein surface expression failed to interact with these GAGs. Soluble M protein bound DS directly and could also inhibit the interaction between DS and S. pyogenes. Experiments with M protein fragments and with streptococci expressing deletion constructs of M protein, showed that determinants located in the NH2-terminal part as well as in the C-repeat region of the streptococcal proteins are required for full binding to GAGs. Treatment with ABC-chondroitinase and HS lyase that specifically remove DS and HS chains from cell surfaces, resulted in significantly reduced adhesion of S. pyogenes bacteria to human epithelial cells and skin fibroblasts. Together with the finding that exogenous DS and HS could inhibit streptococcal adhesion, these data suggest that GAGs function as receptors in M protein-mediated adhesion of S. pyogenes. PMID- 12752451 TI - Crystal structure of thiamindiphosphate-dependent indolepyruvate decarboxylase from Enterobacter cloacae, an enzyme involved in the biosynthesis of the plant hormone indole-3-acetic acid. AB - The thiamin diphosphate-dependent enzyme indolepyruvate decarboxylase catalyses the formation of indoleacetaldehyde from indolepyruvate, one step in the indolepyruvate pathway of biosynthesis of the plant hormone indole-3-acetic acid. The crystal structure of this enzyme from Enterobacter cloacae has been determined at 2.65 A resolution and refined to a crystallographic R-factor of 20.5% (Rfree 23.6%). The subunit of indolepyruvate decarboxylase contains three domains of open alpha/beta topology, which are similar in structure to that of pyruvate decarboxylase. The tetramer has pseudo 222 symmetry and can be described as a dimer of dimers. It resembles the tetramer of pyruvate decarboxylase from Zymomonas mobilis, but with a relative difference of 20 degrees in the angle between the two dimers. Active site residues are highly conserved in indolepyruvate/pyruvate decarboxylase, suggesting that the interactions with the cofactor thiamin diphosphate and the catalytic mechanisms are very similar. The substrate binding site in indolepyruvate decarboxylase contains a large hydrophobic pocket which can accommodate the bulky indole moiety of the substrate. In pyruvate decarboxylases this pocket is smaller in size and allows discrimination of larger vs. smaller substrates. In most pyruvate decarboxylases, restriction of cavity size is due to replacement of residues at three positions by large, hydrophobic amino acids such as tyrosine or tryptophan. PMID- 12752452 TI - Studies on structure-function relationships of indolepyruvate decarboxylase from Enterobacter cloacae, a key enzyme of the indole acetic acid pathway. AB - Enterobacter cloacae, isolated from the rhizosphere of cucumbers, produces large amounts of indole-3-acetic acid. Indolepyruvate decarboxylase, the key enzyme in the biosynthetic pathway of indole-3-acetic acid, catalyses the formation of indole-3-acetaldehyde and carbon dioxide from indole-3-pyruvic acid. The enzyme requires the cofactors thiamine diphosphate and magnesium ions for catalytic activity. Recombinant indolepyruvate decarboxylase was purified from the host Escherichia coli strain JM109. Specificity of the enzyme for the substrates indole-3-pyruvic acid, pyruvic acid, benzoylformic acid, and seven benzoylformic acid analogues was investigated using a continuous optical assay. Stopped-flow kinetic data showed no indication for substrate activation in the decarboxylation reaction of indole-3-pyruvic acid, pyruvic acid or benzoylformic acid. Size exclusion chromatography and small angle X-ray solution scattering experiments suggested the tetramer as the catalytically active state and a pH-dependent subunit association equilibrium. Analysis of the kinetic constants of the benzoylformic acid analogues according to Hansch et al. [Hansch, C., Leo, A., Unger, S.H., Kim, K.H., Nikaitani, D & Lien, E.J. (1973) J. Med. Chem.16, 1207 1216] and comparison with indole-3-pyruvic acid conversion by pyruvate decarboxylases from Saccharomyces cerevisiae and Zymomonas mobilis provided some insight into the catalytic mechanism of indolepyruvate decarboxylase. PMID- 12752453 TI - Covalent and three-dimensional structure of the cyclodextrinase from Flavobacterium sp. no. 92. AB - Starting with oligopeptide sequences and using PCR, the gene of the cyclodextrinase from Flavobacterium sp. no. 92 was derived from the genomic DNA. The gene was sequenced and expressed in Escherichia coli; the gene product was purified and crystallized. An X-ray diffraction analysis using seleno-methionines with multiwavelength anomalous diffraction techniques yielded the refined 3D structure at 2.1 A resolution. The enzyme hydrolyzes alpha(1,4)-glycosidic bonds of cyclodextrins and linear malto-oligosaccharides. It belongs to the glycosylhydrolase family no. 13 and has a chain fold similar to that of alpha amylases, cyclodextrin glycosyltransferases, and other cyclodextrinases. In contrast with most family members but in agreement with other cyclodextrinases, the enzyme contains an additional characteristic N-terminal domain of about 100 residues. This domain participates in the formation of a putative D2-symmetric tetramer but not in cyclodextrin binding at the active center as observed with the other cyclodextrinases. Moreover, the domain is located at a position quite different from that of the other cyclodextrinases. Whether oligomerization facilitates the cyclodextrin deformation required for hydrolysis is discussed. PMID- 12752455 TI - Overview: idiopathic generalized epilepsies. AB - The idiopathic generalized epilepsies (IGEs) are an underemphasized topic. Two reasons for this relative lack of attention are that these epilepsies tend to be more easily controlled than the symptomatic partial and generalized epilepsies, and they are not as common. Because IGE usually arises in childhood or adolescence, these epilepsies may be thought of as a pediatric problem. However, a large number of patients continue to have seizures in adult life. Many exciting developments in understanding the pathophysiology, genetic etiology, and expanded treatment options warrant a reexamination of this important group of the epilepsies. This article reviews the more common IGE syndromes and associated seizure types as the first step in identifying the recent advances in our knowledge of these syndromes. PMID- 12752456 TI - From molecules to networks: cortical/subcortical interactions in the pathophysiology of idiopathic generalized epilepsy. AB - Generalized epilepsy involves abnormally synchronized activity in large-scale neuronal networks. Burst firing of action potentials is a potent mechanism for increasing neural synchrony and is thought to enhance cortical and thalamic rhythmic network activity. Absence seizures, a form of generalized epilepsy, occur in children as brief 5- to 10-s periods of behavioral arrest associated with massive 3- to 4-Hz spike-wave discharges in cortical and thalamic networks. Prior research has shown that enhanced burst firing may be crucial for the transition from normal to epileptic activity. Can enhanced burst firing in one region of the nervous system, such as the cortex, transform the entire thalamocortical network from normal activity to spike-and-wave seizures? Enhanced burst firing in corticothalamic neurons may increase gamma-aminobutyric acid-B (GABAB) receptor activation in the thalamus, leading to the slower, more synchronous oscillations seen in spike-and-wave seizures. Does "generalized" spike-wave activity homogeneously involve the entire brain, or are there crucial nodes that are more important than others for the generation and behavioral manifestations of generalized seizures? Animal and human data suggest that so called generalized seizures involve selective thalamocortical networks while sparing others. A greater understanding of these molecular and network mechanisms will ultimately lead to improved targeted therapies for generalized epilepsy. PMID- 12752457 TI - Exploring new gene discoveries in idiopathic generalized epilepsy. AB - Most epilepsies are categorized under the umbrella term "idiopathic;" these seizure disorders lack a known cause. New genetic technologies are rapidly identifying specific genes responsible for idiopathic generalized epilepsies (IGEs) and are gradually taking the "I" out of "IGE." Ion channel (both voltage- and receptor-mediated) mutations have been linked to a variety of epilepsies considered idiopathic. Gene errors alter excitability in various ways, depending on the mutation, the regional network, and the stage of brain development. The majority of mutations prolong depolarization, favor repetitive firing, and alter neurotransmitter release or postsynaptic sensitivity at central synapses, but the reason for specific seizure types is unclear. Further analyses of these gene mutations and their effects on the developing brain are providing critical clues in the search to explain the origin of "idiopathic" epilepsy. PMID- 12752458 TI - Acute management of seizures in the syndromes of idiopathic generalized epilepsies. AB - Three of the seizure types (myoclonic, absence, and generalized tonic-clonic) and syndromes associated with idiopathic generalized or genetic epilepsies can present an acute status epilepticus picture that requires acute therapy. These are not the usual seizures observed in status epilepticus because most of these patients have secondary generalized or symptomatic generalized convulsive seizures. In this review, I discuss the unique presentation and treatment options for the acute management of seizures in the syndromes of idiopathic generalized epilepsy (IGE), with special emphasis on the seizures of status epilepticus, which persist over time or occur in a series without recovery of consciousness. PMID- 12752459 TI - Chronic management of seizures in the syndromes of idiopathic generalized epilepsy. AB - As a group, idiopathic generalized epilepsies (IGEs) have the highest rates of complete seizure control with medication. However, there are little evidence based data to guide drug choice for treatment. Examples of IGE include absence epilepsy, generalized tonic-clonic epilepsy, and juvenile myoclonic epilepsy. Generalized epilepsies seem to be particularly vulnerable to seizure aggravation, and medications that are primarily effective against partial seizures are more commonly involved in seizure aggravation than other medications. A review of current research has shown that only a few medications can control IGE without potentially causing seizure aggravation. Broad-spectrum antiepileptic drugs such as valproate (VPA), lamotrigine, and topiramate are extremely effective at controlling a variety of seizures without causing excessive seizure aggravation. Among these drugs, VPA has the longest clinical experience history and the largest body of published data. PMID- 12752460 TI - Anticonvulsant effect of flutamide on seizures induced by pentylenetetrazole: involvement of benzodiazepine receptors. AB - PURPOSE: There is some structural similarity between the androgen receptor antagonist, flutamide (Flut) and benzodiazepines (BZDs). We evaluated the possible anticonvulsant effect and interaction of Flut with BZD receptors in common seizure models. METHODS: (a) Different groups of mice each were pretreated i.p. with Flut, and after 0.5 h, they received chemoconvulsants [pentylenetetrazole (PTZ), bicuculline, aminophylline, strychnine or kainic acid]. Latency and incidence of a clonic seizure were recorded. (b) Mice were pretreated i.p. with Flut, and after 0.5 h, transauricular electroshock was applied. Occurrence of a tonic seizure was observed. (c) Amygdala-kindled rats were pretreated i.p. with Flut, and 0.5, 1, or 2 h later, they were stimulated at afterdischarge threshold. Then the seizure parameters (afterdischarge duration, seizure severity, and stage 5 duration) were recorded. (d) The effect of Flut on clonic seizure threshold was determined by i.v. infusion of bicuculline or PTZ to different groups of Flut-receiving mice. To determine the possible interaction of Flut with BZD receptors, the flumazenil (FMZ)+Flut effect on clonic seizure threshold was compared with the effect of Flut. (e) Neurotoxicity of Flut was evaluated by rotarod test at 30 min after administration. RESULTS: Flut produced a dose-dependent anticonvulsant effect against PTZ-induced seizures [median effective dose (ED50), 67.0 mg/kg]. Moreover, Flut elevated the clonic seizure threshold induced by bicuculline or PTZ. FMZ reversed the effect of Flut on the threshold of PTZ seizures. A median toxic dose (TD50) value of 124.8 mg/kg was obtained for Flut. CONCLUSIONS: Flut both blocks PTZ-induced clonic seizures and elevates the threshold of PTZ or bicuculline-induced clonic seizures, through interaction with BZD receptors. PMID- 12752461 TI - Short bursts of weak pulses break postictal inhibition in the neocortex of Wistar rats. AB - PURPOSE: Postictal inhibition (PI) is a decrease in excitability that follows an epileptic seizure and decreases probability of new seizure occurrence. PI may involve both increased inhibition and persisting elevated excitation. Our experiments tested whether shorter trains of weak stimuli are able to unmask this residual increase of excitability during the PI. METHODS: Four epileptic afterdischarges (ADs) were evoked by intense electrical stimulation (20 s, 8 Hz, current intensity at 5x threshold) of the neocortex in two groups (A, B) of Wistar rats. Before the first AD and during the 10-min interictal period, 8-Hz trains of four weak pulses (half of the intensity used for the AD triggering; 4P) were applied every 20 s in group B and a single pulse with similar parameters in group A. RESULTS: The number of interictal epileptiform events evoked by 4P in the group B was significantly higher than that in the group A (evoked by single pulses) except after the second AD. Epileptic events were triggered by 4P also immediately after the AD termination. CONCLUSIONS: It is apparent that weak stimulation can trigger epileptic phenomena during PI. Our results indicate that it is no longer possible to perceive PI only as persisting extreme and active inhibition. An appropriate stimulation can reveal more subtle (but important) excitatory events contributing to the functional status during the postictal period. PMID- 12752462 TI - Effect of topiramate on the anticonvulsant activity of conventional antiepileptic drugs in two models of experimental epilepsy. AB - PURPOSE: The objective of this study was to evaluate the interaction of the novel antiepileptic drug (AED), topiramate (TPM), with conventional AEDs against amygdala-kindled seizures in rats and pentylenetetrazol-induced convulsions in mice. METHODS: Experiments were performed on mice and fully kindled rats. In pentylenetetrazol test, the chemoconvulsant was used at its CD97 dose of 105 mg/kg, producing clonic seizures in 97% of mice. Adverse effects were evaluated with the chimney test and passive avoidance task. Plasma levels of AEDs were measured with immunofluorescence. RESULTS: TPM at 20 mg/kg exerted a significant anticonvulsant effect as regards seizure and afterdischarge durations in amygdala kindled seizures in rats, being ineffective at lower doses. Coadministration of TPM (10 mg/kg) with valproate (VPA; at a subtherapeutic dose of 50 mg/kg) resulted in essential reductions of seizure and afterdischarge durations. TPM (10 mg/kg) combined with carbamazepine (CBZ; at a subtherapeutic dose of 15 mg/kg) significantly increased afterdischarge threshold, simultaneously decreasing the remaining seizure parameters (duration or severity of seizures and afterdischarge duration). TPM (10 mg/kg) given with phenobarbital (PB; 15 mg/kg) markedly shortened seizure severity and seizure and afterdischarge durations. Combinations of TPM with diphenylhydantoin (PHT) were ineffective against kindled seizures in rats. TPM combined with VPA and PB did not alter their plasma levels, but its combination with CBZ resulted in an increased free plasma CBZ concentration. TPM (10 and 20 mg/kg) alone and its combinations with conventional AEDs affected neither motor coordination nor long-term memory, evaluated in the chimney and passive avoidance tests, respectively, in rats. In pentylenetetrazol-evoked convulsions in mice, TPM (175 and 200 mg/kg) showed anticonvulsant effects per se. Moreover, TPM (at its subtherapeutic dose of 150 mg/kg), significantly potentiated the anticonvulsant action of ethosuximide (ESM), but not that of VPA, PB, or clonazepam (CZP) against pentylenetetrazol-induced seizures. Either TPM alone (150 mg/kg) or its combination with ESM did not result in significant undesired effects. CONCLUSIONS: The experimental data indicate that except for PHT, the combinations of TPM with conventional AEDs are beneficial against amygdala-kindled seizures in rats. In the pentylenetetrazol test, this novel AED potentiated only the protection offered by ESM. PMID- 12752463 TI - Neuronal cell death in a rat model for mesial temporal lobe epilepsy is induced by the initial status epilepticus and not by later repeated spontaneous seizures. AB - PURPOSE: To determine whether repeated seizures contribute to hippocampal sclerosis, we investigated whether cell loss in the (para) hippocampal region was related to the severity of chronic seizure activity in a rat model for temporal lobe epilepsy (TLE). METHODS: Chronic epilepsy developed after status epilepticus (SE) that was electrically induced 3-5 months before. The presence of neuronal damage was assessed by using Fluoro-Jade and dUTP nick end-labeling (TUNEL) of brain sections counterstained with Nissl. RESULTS: We found a negative correlation between the numbers of surviving hilar cells and the duration of the SE (r = -0.66; p < 0.01). In the chronic phase, we could discriminate between rats with occasional seizures (0.15 +/- 0.05 seizures per day) without progression and rats with progressive seizure activity (8.9 +/- 2.8 seizures/day). In both groups, the number of TUNEL-positive cells in parahippocampal regions was similar and higher than in controls. In the hippocampal formation, this was not significantly different from controls. Fluoro Jade staining showed essentially the same pattern at 1 week and no positive neurons in chronic epileptic rats. CONCLUSIONS: Cell death in this rat model is related to the initial SE rather than to the frequency of spontaneous seizures. These results emphasize that it is of crucial importance to stop the SE as soon as possible to prevent extended cell loss and further progression of the disease. They also suggest that neuroprotectants can be useful during the first week after SE, but will not be very useful in the chronic epileptic phase. PMID- 12752464 TI - Topiramate and psychiatric adverse events in patients with epilepsy. AB - PURPOSE: The aim of this study was to determine the prevalence of psychiatric adverse events (PAEs) in patients with epilepsy treated with topiramate (TPM). Classification, relation to TPM dosing, and outcome were evaluated to identify a patient profile at risk of developing PAEs. METHODS: We evaluated the data of the first consecutive and prospectively collected patients in therapy with TPM. RESULTS: Follow-up information was available for 431 patients. PAEs occurred in 103 (23.9%) patients; M/F ratio, 55:48; mean age (+/-SD), 36.5 +/- 11.2. In 46 (10.7%) patients, an affective disorder developed; in 16 (3.7%), a psychotic disorder; in 24 (5.6%), aggressive behavior with or without irritability; in 17 (3.9%), other behavior abnormalities such as agitated behavior, anger/hostility behavior, or anxiety. High starting dose and rapid titration schedule were relevant for the development of PAEs. Family psychiatric history and family history of epilepsy, personal history of febrile convulsions, psychiatric history, and presence of tonic-atonic seizures were found to be significant risk factors. Low seizure frequency before starting TPM and TPM/lamotrigine coadministration had a protective effect for PAEs. CONCLUSIONS: We found that PAEs associated with TPM were related to the titration schedule of the drug and that a unique patient profile is suggested by the clinical history. PMID- 12752465 TI - Statistical mapping of scalp-recorded ictal EEG records using wavelet analysis. AB - PURPOSE: The wavelet transform (WT) is well suited for the analysis of signals whose characteristics vary rapidly over time. We devised a computerized method for objective scoring of scalp-recorded seizures that takes advantage of the WT. METHODS: Using wavelet coefficients as a metric, we devised a statistical scoring method aimed at detecting significant and sustained rhythmic buildup. The approach was used to create spatiotemporal significance maps for each seizure. Each seizure was also independently analyzed by computer and an expert reader not involved in the clinical workup or computer analysis of these patients. Hierarchical decision rules for determining seizure lateralization and localization were established from a training set of seizures and subsequently tested on those from an independent test set of seizures. The test dataset included a total of 57 scalp-recorded seizures from 18 patients, each with a > or =12-month seizure-free surgical outcome. RESULTS: Validation was determined by the site of surgical resection. Of the 57 seizure records in the test dataset, the computerized approach resulted in 48 correctly lateralized seizures as compared to 34 for the expert reader. Further, the computer correctly localized 41 seizures to the expert's 31. CONCLUSIONS: The method presented appears to provide an objective basis for the intrachannel scoring of ictal EEGs with minimal interference from artifacts and intermittent discharges. Although the approach has so far shown a substantial improvement over expert scoring in estimating the lateralization and locus of seizure onset, further testing is required to fully evaluate fully its diagnostic accuracy. PMID- 12752466 TI - A prospective study of the requirement for and the provision of epilepsy surgery in the United Kingdom. AB - PURPOSE: Of the 30,000 persons in whom epilepsy develops annually in the United Kingdom, in approximately 6000 (20%), intractability develops. Some of these patients will be appropriate for epilepsy surgery. We aimed to estimate the number of patients who should be considered surgical candidates, by extrapolation from a population-based study of prognosis and the number who are receiving epilepsy surgery, by a survey of U.K. neurosurgeons. METHODS: We identified the number of patients who may eventually require surgery from a prospective cohort of patients with newly diagnosed epilepsy. We identified all U.K. neurosurgeons who had performed any epilepsy surgery in the past year. Each identified surgeon prospectively recorded the number and types of operations carried out for 6 months. RESULTS: Of newly diagnosed patients each year, 450 (1.5%) may eventually require surgery. Thirty-two respondents (22% of all U.K. neurosurgeons) reported that they performed epilepsy surgery. The 211 operations were carried out in the 6 months surveyed (422 operations annually or 13 per surgeon per year). Temporal lobe resection (77%) was the most common procedure. CONCLUSIONS: Based on a prevalence of 5/1,000 persons with epilepsy, < or =4,500 patients in the U.K. require epilepsy surgery. Every year, 450 patients with newly diagnosed epilepsy who may eventually require surgery are added to this "surgical pool." At the current annual rate of operations, a large number of refractory patients remain untreated. This is probably partly because many patients are not referred for specialist care and therefore remain underinvestigated. PMID- 12752467 TI - A retrospective analysis of hippocampal pathology in human temporal lobe epilepsy: evidence for distinctive patient subcategories. AB - PURPOSE: This study is a retrospective analysis of the pathology of the hippocampus from patients with medically intractable temporal lobe epilepsy. We attempted to relate neuronal density, immunohistochemistry, electrophysiologic data, and surgical outcome. METHODS: Immunostaining patterns for neuropeptide Y, somatostatin, substance P, and dynorphin defined the immunohistochemical characteristics of the hippocampi. Neuronal densities were determined by microscopic cell counts. Sharp electrode recordings from dentate granule cells determined measures of inhibition and excitation. RESULTS: Patient hippocampi without evidence of sclerosis generally resembled autopsy controls on the basis of neuronal densities of hippocampal subfields and patterns of immunostaining. The nonsclerotic hippocampi were divisible into two subgroups on the basis of neuronal density correlations between hippocampal subfields, the excitability of dentate granule cells, etiology, and surgical outcome. Hippocampi with sclerosis were divisible into those with significant neuronal loss confined to area CA1 and those with neuronal loss throughout the hippocampus and dentate gyrus. In the former, the dentate gyrus resembled in morphology the nonsclerotic hippocampi but with slightly increased excitability of the dentate granule cells. The hippocampi with more extensive neuronal loss had changes in immunostaining patterns associated with the dentate gyrus, correlated with significant hyperexcitability of dentate granule cells. The surgical outcome, with the exception of one group, was good in approximately 70-90%. CONCLUSIONS: Hippocampi from patients with intractable temporal lobe epilepsy can be assigned to several groups on the basis of pathophysiology. Different pathologies may represent differing causative mechanisms of intractable temporal lobe epilepsy and be predictive of surgical outcome. PMID- 12752468 TI - EEG-related functional MRI in benign childhood epilepsy with centrotemporal spikes. AB - The localization of epileptic foci is an important issue in children with extratemporal epilepsies. However, the value of noninvasive methods such as the EEG-assisted functional magnetic resonance imaging (fMRI) has not been sufficiently investigated in children. As a model of extratemporal epilepsies, we studied 7 patients aged 5 to 12 (median 10) years with benign childhood epilepsy and centrotemporal (rolandic) spikes. Interictal spikes were recorded during the fMRI acquisition on a MR-compatible battery-powered digital EEG system with 16 channels. The fMRI sequences were correlated off-line with the EEG spikes and analyzed with the software Statistical Parametrical Mapping SPM99. The fMRI results demonstrated the spike-related activation in the perisylvian central region in three patients; we could not demonstrate fMRI activation despite active spiking in 2 patients, and 2 patients did not produce sufficient spikes for fMRI analysis. We currently consider the spike-related fMRI as a research tool that localizes epileptic activity in selected patients. Further improvements of the technique are necessary to allow a clinical application of this method. PMID- 12752469 TI - Epileptic spasms and partial seizures as a single ictal event. AB - PURPOSE: To investigate the phenomenon of epileptic spasms (ESs) associated with other seizure types in a single ictal event and to study the predictive value of this phenomenon regarding etiology and prognosis. METHODS: We selected retrospectively eight female and five male patients, who had ESs and other seizure types within a single seizure event and for whom a video-EEG recording of the phenomenon was performed in at least one situation. RESULTS: The seizure type associated with ESs was a partial seizure in all patients. We identified three groups with different seizure patterns regarding the temporal association of ES and partial seizures (PSs): (a) PS followed by ES; (b) PS appearing during a cluster of ESs without interrupting the cluster; and (c) complex seizure interaction with a succession of ESs and PSs in a close but variable temporal association. Underlying disorders included cortical dysplasia (three patients), complex cerebral malformations (two patients), and perinatal anoxic-ischemic injuries (two patients); four cases were classified as cryptogenic, and in two children, etiology was unknown, but prenatal origin was suspected. Outcome was poor in nine cases with intractable epilepsy; four cases had a favorable outcome, defined as complete cessation of epileptic seizures. CONCLUSIONS: The phenomenon of associated ESs and PSs as a single ictal event can be related to different etiologies and should not be considered distinctive for cortical malformations or severe brain damage. Different seizure patterns of associated ESs and PSs provide no hint for etiology or prognosis. Outcome is prevalently but not constantly unfavorable in patients with the phenomenon. PMID- 12752470 TI - Seizure characterization and electroencephalographic features in Glut-1 deficiency syndrome. AB - PURPOSE: To characterize seizure types and electroencephalographic features of glucose transporter type 1 deficiency syndrome (Glut-1 DS). METHODS: Twenty children with clinical and laboratory features of Glut-1 DS were evaluated. Age at seizure diagnosis, seizure classification, and response to treatment were determined by chart review. Thirty-two continuous 24-h EEG monitoring sessions and reports of 42 routine EEG studies were assessed. RESULTS: Age at seizure diagnosis was between 4 weeks and 18 months (mean, 5 months). Seizure types were generalized tonic or clonic (14), absence (10), partial (nine), myoclonic (six), or astatic (four). During 24-h EEGs, background activity showed generalized 2.5- to 4-Hz spike-wave discharges (41%), generalized slowing or attenuation (34%), no abnormalities (34%), focal epileptiform discharges (13%), or focal slowing or attenuation (9%). No seizures were captured during 69% of the studies; the remainder had absence (19%), myoclonic (9%), or partial seizures (3%). On evaluation of routine and 24-h EEG studies, focal epileptiform discharges (24%) and slowing (11%) were more frequent in ages 0-24 months. In older children (2-8 years), generalized epileptiform discharges (37.5%) and slowing (21%) were more common. CONCLUSIONS: In all ages, a normal interictal EEG was the most common EEG finding. When abnormalities occurred, focal slowing or epileptiform discharges were more prevalent in the infant. In older children (2 years or older), a generalized 2.5- to 4-Hz spike-wave pattern emerged. Seizure types observed included, absence, myoclonic, partial, and astatic. PMID- 12752471 TI - Effects of valproic acid, carbamazepine, and phenobarbitone on the fatty acid composition of erythrocyte membranes in children. AB - PURPOSE: To investigate a possible relation between antiepileptic drugs (AEDs) and the fatty acid composition of membranes. METHODS: Fatty acid (FA) composition of erythrocytes was studied in children with epilepsy receiving AED monotherapy. Children taking valproate (VPA, n = 28), carbamazepine (CBZ, n = 17), or phenobarbitone (PB, n = 14) were compared with healthy controls (n = 25). FAs were measured by capillary-gas chromatography (GC-FID). RESULTS: Significant changes (p < 0.05) in the FA composition of membranes were found. In children treated with VPA, C13:0 was decreased (8.2 +/- 2% vs. 10.7 +/- 4% in controls) and C14:0 increased (1.4 +/- 0.5% vs.1 +/- 0.5% in controls). C17:0 again was reduced (9.9 +/- 4% vs. 13.2 +/- 6% in controls), whereas the long-chained acids were enhanced: C18:2n-6 (6 +/- 2.4% vs. 3.9 +/- 2.5% in controls), and C20:4n-6 (1.9 +/- 1.7% vs. 1.4 +/- 0.5% in controls). The nonidentified FAs also increased with VPA therapy: 2.5 +/- 0.8% versus 1.7 +/- 0.9% in controls. Children treated with CBZ showed only minimal changes of FA composition: C13:0 was decreased compared with controls (8 +/- 2% vs. 10.7 +/- 4%). No changes were seen in patients taking PB. The mean corpuscular volume (MCV) showed important differences between the study groups: MCV was 84.7 +/- 6.0 fl during VPA therapy (p < 0.001) and 85.7 +/- 4.1 fl with CBZ (p < 0.001). During PB, the MCV increased to 82.87 +/- 3.29 fl compared with controls (78.73 +/- 4.92 fl; p < 0.01). CONCLUSIONS: VPA therapy is associated with changes of the FA composition of membranes, which is not the case with PB therapy. The implications of this finding remain to be established. PMID- 12752472 TI - Respiratory pattern in a rat model of epilepsy. AB - PURPOSE: Apnea is known to occur during seizures, but systematic studies of ictal respiratory changes in adults are few. Data regarding respiratory pattern defects during interictal periods also are scarce. Here we sought to generate information with regard to the interictal period in animals with pilocarpine-induced epilepsy. METHODS: Twelve rats (six chronically epileptic animals and six controls) were anesthetized, given tracheotomies, and subjected to hyperventilation or hypoventilation conditions. Breathing movements caused changes in thoracic volume and forced air to flow tidally through a pneumotachograph. This flow was measured by using a differential pressure transducer, passed through a polygraph, and from this to a computer with custom software that derived ventilation (VE), tidal volume (VT), inspiratory time (TI), expiratory time (TE), breathing frequency (f), and mean inspiratory flow (VT/TI) on a breath-by-breath basis. RESULTS: The hyperventilation maneuver caused a decrease in spontaneous ventilation in pilocarpine-treated and control rats. Although VE had a similar decrease in both groups, in the epileptic group, the decrease in VE was due to a significant (p < 0.05) increase in TE peak in relation to that of the control animals. The hypoventilation maneuver led to an increase in the arterial Paco2, followed by an increase in VE. In the epileptic group, the increase in VE was mediated by a significant (p < 0.05) decrease in TE peak compared with the control group. Systemic application of KCN, to evaluate the effects of peripheral chemoreception activation on ventilation, led to a similar increase in VE for both groups. CONCLUSIONS: The data indicate that pilocarpine-treated animals have an altered ability to react to (or compensate for) blood gas changes with changes in ventilation and suggest that it is centrally determined. We speculate on the possible relation of the current findings on treating different epilepsy-associated conditions. PMID- 12752473 TI - Surgical treatment of multifocal epilepsy involving eloquent cortex. AB - PURPOSE: This report describes our long-term follow-up for combined resective surgery and multiple subpial transections (MSTs) in patients with refractory epilepsy involving eloquent and noneloquent cortex in multiple lobes. Multiple independent seizure foci made these patients poor candidates for conventional surgery. METHODS: MST and resective surgery were used in 13 patients to treat localization-related refractory epilepsy involving eloquent and noneloquent cortex of two or more lobes. Preoperative investigation was followed by invasive monitoring. RESULTS: Eleven patients had MST plus resection involving two different lobes, and two patients had MST plus resection involving three different lobes. MSTs were performed on the primary sensorimotor cortex (eight patients), temporal language area (two patients), Broca's area (one patient), and on both frontal motor and temporal language areas (two patients). Nine patients had a two-stage procedure, and four patients had a three-stage procedure (two consecutive subdural grid studies followed by resections). Average follow-up was 59.2 months (range, 42-98 months). With a modified Engel Outcome Scale, four patients (31%) had a class I outcome; three (23%), class II; three (23%), class III; and three (23%), class IV. Ten (77%) patients had a >50% reduction of seizure burden. CONCLUSIONS: Combined MST and resection can meaningfully improve seizure control in patients with multifocal epilepsy involving eloquent cortex. Prospective randomized studies are needed. PMID- 12752474 TI - Safety and efficacy of intravenous valproate in pediatric status epilepticus and acute repetitive seizures. AB - PURPOSE: To evaluate the safety and efficacy of intravenous valproate (VPA) loading in children with status epilepticus (SE) or acute repetitive seizures. METHODS: Retrospective review was performed on 40 pediatric patients with intravenous VPA loading. Patients were classified into two groups: SE (n = 18) and acute repetitive seizures (n = 22). Thirty-one patients were VPA naive and received a full loading dose of 25 mg/kg; nine had subtherapeutic plasma VPA levels and received a partial loading dose. Average infusion rate was 2.8 mg/kg/min. Heart rate and blood pressure were measured before, during, and after infusion. RESULTS: Intravenous VPA loading stopped seizures in 18 patients with SE within 20 min. All 18 patients regained baseline mental status within 1 h of seizure cessation. Among 22 patients with acute repetitive seizures, only one had further seizures after VPA infusion. One patient in the SE group complained of transient tremors. No significant changes in blood pressure or heart rate were found in either group. Postinfusion plasma VPA levels ranged from 51 to 138 microg/ml (mean +/- SD = 88 +/- 21.5 microg/ml). CONCLUSIONS: Intravenous VPA loading is safe and effective for treating acute seizure emergencies in children. PMID- 12752475 TI - ILAE Commission of European Affairs Subcommission on European Guidelines 1998 2001: The provision of epilepsy care across Europe. AB - PURPOSE: To assess the needs and resources available in the provision of basic epilepsy care across Europe. METHODS: A mailed questionnaire was used, the European Epilepsy Services inventory (EESI). The EESI was distributed to all 36 European chapters of the International League Against Epilepsy (ILAE), and answers were obtained from 32, a response rate of 89%. For the purpose of studying trends across Europe, the chapters were divided into a Western, an Eastern, a Central, and a Southern group. RESULTS: The survey results showed that there was a wide range in the number of physicians and specialists involved in epilepsy care across Europe, with a trend toward higher numbers of neurologists, pediatricians, and pediatric neurologists in Eastern Europe. Many different specialties were involved in epilepsy care, and many chapters reported differences in the provision of care across their countries, with less possibility for patients to see a specialist in the least provided areas, where most epilepsy patients were cared for by general practitioners and internists. Problems with high costs of the newer antiepileptic drugs were most pronounced in Eastern Europe. Problems with lack of comprehensive care and of epilepsy specialists, with stigma and social problems, and with insufficient professional education and knowledge about epilepsy were reported all across Europe. CONCLUSIONS: Knowledge about differences in the pattern of provision of epilepsy care and about the main problems encountered by the European ILAE chapters is of importance in the continuing efforts to improve management of epilepsy all over Europe. PMID- 12752476 TI - The new proposal on classification of epileptic seizures and epilepsy syndromes. PMID- 12752477 TI - Utility of sphenoidal electrodes for review of ictal tracings. PMID- 12752479 TI - Unsuspected atypical hemispheric dominance for language as determined by FMRI. Handedness: neural versus behavioral: the difference is measurable. PMID- 12752480 TI - Kidney stones, carbonic anhydrase inhibitors, and the ketogenic diet. PMID- 12752481 TI - Is birth weight related to later glucose and insulin metabolism?--A systematic review. AB - AIM: To determine the relationship of birth weight to later glucose and insulin metabolism. METHODS: Systematic review of the published literature. Data sources were Medline and Embase. Included studies were papers reporting the relationship of birth weight with a measure of glucose or insulin metabolism after 1 year of age, including the prevalence of Type 2 diabetes mellitus (DM). Three reviewers abstracted information from each paper according to specified criteria. RESULTS: Forty-eight papers fulfilled the criteria for inclusion, mostly of adults in developed countries. Most studies reported an inverse relationship between birth weight and fasting plasma glucose concentrations (15 of 25 papers), fasting plasma insulin concentrations (20 of 26), plasma glucose concentrations 2 h after a glucose load (20 of 25), the prevalence of Type 2 DM (13 of 16), measures of insulin resistance (17 of 22), and measures of insulin secretion (16 of 24). The predominance of these inverse relationships and the demonstration in a minority of studies of other directions of the relationships could not generally be explained by differences between studies in the sex, age, or current size of the subjects. However, the relationship of birth weight with insulin secretion was inconsistent in studies of adults. CONCLUSIONS: The published literature shows that, generally, people who were light at birth have an adverse profile of later glucose and insulin metabolism. This is related to higher insulin resistance, but the relationship to insulin secretion in adults is less clear. PMID- 12752482 TI - Amniotic fluid insulin levels and fetal abdominal circumference at time of amniocentesis in pregnancies with diabetes. AB - AIMS: Fetal hyperinsulinism is a strong predictor for excessive growth and fetopathy in pregnancies complicated by diabetes. We examined (i). the relationship between measurements of amniotic fluid insulin (AF insulin) and fetal abdominal circumference (AC) at the time of amniocentesis, and (ii). whether there is a threshold for fetal AC percentiles which can identify low vs. high-risk levels of AF insulin without performing an amniocentesis. METHODS: In a retrospective study, AF insulin from 121 pregnant diabetic women (32 pregestational; 89 gestational) was measured during the 3rd trimester as part of a diabetes management protocol. AC measurements were transformed into a continuous variable of percentile growth for gestational age (Hadlock). Division of the cohort according to deciles or quartiles of AC percentiles was performed to identify a threshold AC with a significant increase in elevated AF insulin, previously defined as AF insulin >or= 7 microU/ml. A receiver operator characteristic (ROC) curve was created and the negative predictive value (NPV) of the determined threshold was calculated. RESULTS: AF insulin levels were significantly correlated with the AC percentiles (r = 0.3, P = 0.0005) by linear regression. No AC threshold could reliably identify a moderate elevated AF insulin >or= 7 microU/ml (NPV 77.2%), but an AC threshold >or= 75th percentile could identify with fetal hyperinsulinism with an AF insulin >or= 16 microU/ml. All 10 cases of AF insulin >or= 16 microU/ml were identified with a NPV of 100% (74/74). CONCLUSIONS: Our data indicate that an AC >or= 75th percentile determined by a 3rd trimester ultrasound examination may discriminate between pregnancies at low vs. high risk for AF insulin >or= 16 microU/ml. This AF insulin concentration corresponds to a level of hyperinsulinism reported to be associated with considerable neonatal and long term morbidity. PMID- 12752483 TI - How do people with Type 2 diabetes perceive and respond to cardiovascular risk? AB - OBJECTIVES: To explore how people with Type 2 diabetes perceive cardiovascular risk, and how those perceptions might affect their motivation to make lifestyle changes. METHODS: The setting was a diabetes clinic in a UK teaching hospital. A qualitative study was conducted, using semistructured individual interviews and template analysis of content. The participants were 20 Type 2 diabetic patients, aged between 52 and 77 years, half with and half without cardiovascular disease (CVD). RESULTS: Whether they had CVD or not, most people were aware they were at risk of it, of its causative factors, and possible effects. However, they were more likely to attribute it to unchangeable factors like 'stress' and 'heredity', than medical risk factors like cholesterol and smoking. Patients with pre existing CVD correctly regarded their risk as higher than those without. Few saw any direct link between being diabetic per se and cardiovascular risk. Lifestyle changes were precipitated by major life events and motivated by family support, fear of complications, and a belief that one should follow doctors' advice. A common reaction to CVD and diabetes was stoical acceptance, allowing patients to view their lives positively, whilst living with unpredictable potentially disabling diseases. CONCLUSIONS: Patients were unaware how strongly diabetes influences cardiovascular risk. Their ideas about risk were very different from those of conventional medicine, and provided individual rationales for making choices about treatment and risk-influencing behaviour. Contextual factors, such as family milieu, also influenced their behaviour. Clinicians should not assume patients share the same mental model of risk as they, and must be prepared to explore peoples' individual constructs and health beliefs. PMID- 12752484 TI - Proton magnetic resonance spectroscopy study of soleus muscle in non-obese healthy and Type 2 diabetic Asian Northern Indian males: high intramyocellular lipid content correlates with excess body fat and abdominal obesity. AB - AIMS: Intramyocellular lipids (IMCL) appears to be important in the pathogenesis of insulin resistance. Correlation of IMCL content of soleus muscle with insulin sensitivity has been reported in the Caucasian population. In the present study, IMCL content was estimated in the soleus muscle of both non-obese healthy males and Type 2 diabetic males, and correlated with the anthropometric parameters, blood glucose, plasma lipids, and insulin resistance in Asian Indians from North India. METHODS: Twenty males (Type 2 diabetes mellitus 10; healthy controls 10) with body mass index (BMI) 25. The following were assessed in all subjects: body composition, fasting blood glucose, lipid profile, insulin levels, insulin resistance by homeostasis model assessment, and proton magnetic resonance spectroscopy (1H MRS) study of the soleus muscle. RESULTS: IMCL content was approximately two times higher in Type 2 diabetic males compared with healthy males (P < 0.05). Amongst healthy males, IMCL content was significantly higher (P < 0.05) in subjects with percentage BF > 25 compared with subjects with percentage BF or= 25. Similarly, IMCL content was high in subjects with waist-hip ratio (WHR) > 0.95 compared with subjects with WHR 3 did not exhibit a significant increase in MBF in response to CPT. After administration of deferoxamine, the increase in MBF in response to CPT was 23% and the magnitude of increase was related to the MIBG uptake score (r = 0.40, P < 0.0001). CONCLUSIONS: Myocardial blood flow response to sympathetic stimulation is significantly impaired in long-term Type 1 diabetes. After preinfusion with deferoxamine the impairment is partially reversed and a relationship between myocardial blood flow and the extent of cardiac sympathetic dysfunction is observed. PMID- 12752487 TI - Maternal ante-natal parameters as predictors of persistent postnatal glucose intolerance: a comparative study between Afro-Caribbeans, Asians and Caucasians. AB - AIMS: To measure the prevalence of persistent glucose intolerance at 6-12 weeks postpartum in various ethnic groups to assess the value of targeted postpartum screening. METHODS: A retrospective study was performed using computerized databases from two large maternity units within one UK region. Both units used the same screening strategy for gestational diabetes mellitus (GDM) and the same postpartum follow-up at 6-12 weeks using a 75-g oral glucose tolerance test (OGTT). A total of 221 women with a diagnosis of GDM/impaired glucose tolerance (IGT) in the index pregnancy in addition to a completed postpartum 75-g OGTT were studied. Of these, 91 were Caucasian, 89 were of Indo-Asian (Asian) origin and 41 were of Afro-Caribbean origin. RESULTS: The study showed that 35% Indo-Asians had persistent postpartum glucose intolerance compared with 7% Caucasians and 5% Afro Caribbeans (P < 0.003). Insulin requirement during pregnancy and a diagnosis of gestational diabetes prior to 20 weeks of pregnancy were predictive for persistent postpartum glucose intolerance amongst Indo-Asians. CONCLUSIONS: The frequency of postpartum glucose intolerance among Indo-Asian women is significantly greater than among age-matched Caucasian and Afro-Caribbean women. We suggest that all Indo-Asian women with gestational diabetes should undergo postpartum screening for persistent glucose intolerance. However, in non-Asian women selective screening may be more cost effective. PMID- 12752488 TI - Rosiglitazone in Type 2 diabetes mellitus: an evaluation in British Indo-Asian patients. AB - AIMS: To evaluate the effectiveness of rosiglitazone in reducing hyperglycaemia in patients with Type 2 diabetes mellitus (DM) of Indo-Asian origin taking concurrent sulphonylurea therapy. METHODS: A randomized, double-blind, placebo controlled study of 26 weeks' duration at 31 primary and secondary care centres in areas of the UK with a high Indo-Asian population, including 177 patients aged 28-78 years. Rosiglitazone 8 mg/day or matching placebo was added to existing sulphonylurea therapy. The primary endpoint was change from baseline in glycosylated haemoglobin A1c (HbA1c) at week 26. RESULTS: The mean changes in HbA1c were -1.16% with rosiglitazone (baseline 9.21%) and +0.26% with placebo (baseline 9.06%) (treatment difference P < 0.001; 95% confidence interval (CI) 1.81, -1.08). HbA1c fell below 8% in 55% and 19% of patients, respectively (treatment difference P < 0.001; 95% CI 0.22, 0.51). The greatest improvements occurred in patients whose glycaemic control was initially poor. Improvements in homeostasis model assessment of insulin sensitivity and pancreatic beta-cell function with rosiglitazone were not accompanied by a change in plasma insulin or C-peptide after 26 weeks. Free fatty acids fell by 0.09 mmol/l with rosiglitazone and increased by 0.03 mmol/l with placebo (treatment difference P < 0.001; 95% CI -0.19, -0.07). CONCLUSION: Rosiglitazone improved insulin sensitivity, pancreatic beta-cell function, and glycaemic control in Indo-Asian patients with Type 2 DM who are at greater risk of the complications of Type 2 DM than other ethnic groups. PMID- 12752489 TI - An assessment of care of paediatric and adolescent patients with diabetes in a large district general hospital. AB - AIMS: To assess the process of clinical care and outcomes of young patients with diabetes attending clinics at a large district general hospital. METHODS: Retrospective analysis of data obtained from 106 case notes of patients aged 12 22 years attending the paediatric, combined adolescent or adult diabetes clinics between 1998 and 2000. The frequency of follow-up, rate of admission, glycaemic control, systolic blood pressure, weight change and screening for complications were assessed. RESULTS: The mean attendance rate was 78%. The admission rate was 91 admissions per 1000 patient years. Overall, the mean HbA1c was 9.1% with only 15% of paediatric and adolescent patients having mean HbA1c 0.05). Dental trauma was associated with increased touch thresholds in permanent incisor teeth, with recovery toward healthy control values usually occurring between 3-12 months. PMID- 12752538 TI - External and occlusal trauma to dental implants and a case report. AB - Dental implants subjected to traumatic forces can survive. Cortical bone seems to provide a protective energy-absorbing mechanism in the collagen polymer that helps to prevent microcracking and fracture of bone. The collagen polymer has cross-linking bonds that break and absorb the energy of a traumatic impact so as not to cause damage to the main polymer chain. A case reported demonstrates that a traumatic force damaged the implant prosthetic crown, but not the bone encasing the implant or the integration of the implant. PMID- 12752539 TI - Treatment of a non-vital immature incisor with mineral trioxide aggregate (MTA). AB - - A case of severe crown fracture and luxation in the upper permanent incisors of a 9-year-old boy is reported. The treatment of one of the injured teeth included apexification with calcium hydroxide and endodontic treatment with gutta-percha obturation. The other incisor was also treated with calcium hydroxide, but as there was no apical stop after 3 years of treatment, it was decided to use a new root-end filling material: mineral trioxide aggregate (MTA). At follow-up 12 months later, the tooth was asymptomatic and radiographically showed the initial repair of the radiolucent apical lesion. PMID- 12752540 TI - External inflammatory and replacement resorption of luxated, and avulsed replanted permanent incisors: a review and case presentation. AB - External inflammatory resorption and replacement resorption are complicating factors that may result from traumatic dental injuries when the tooth is luxated or avulsed and replanted. Resorption may, ultimately, result in loss of the tooth. However, with appropriate treatment, the prognosis for these teeth is greatly improved, with the possibility of preventing or arresting resorption. The purpose of this paper is to review these trauma entities, to discuss factors that influence the occurrence of resorption and to describe the most appropriate treatment. A case is presented, illustrating both resorption entities, but with varying outcomes. PMID- 12752541 TI - Oral and general health-related quality of life with conventional and implant dentures. AB - Implant overdentures and conventional prostheses have been compared in several trials using a variety of functional and oral health-related quality of life (OHQOL) outcomes. In this paper, we describe the impact of implant overdentures on general and OHQOL in seniors. OBJECTIVES: To compare the oral health-related and general quality of life of seniors (aged 65-75 years) who received either mandibular implant overdentures or conventional dentures. METHODS: Sixty edentulous patients were recruited. Thirty received mandibular overdentures retained by two implants (IOD) and a conventional maxillary denture, the other 30 subjects received new maxillary and mandibular conventional complete dentures (CD). All completed the 20-item version of the Oral Health Impact Profile (OHIP 20) before treatment, then at two and 6 months after delivery of the dentures. The SF-36 general health questionnaire was completed at baseline and 6 months only. RESULTS: Pretreatment and 6-month data from 55 subjects were analyzed. Those who received the IODs had significantly better OHIP-20 total scores at 6 months. Results for IOD subjects were also superior in the functional limitation, physical pain, physical disability and psychological disability subscales. While no significant between group difference was found on the SF-36 health survey, significant pre-post-treatment differences within the IOD group were detected for the role emotional, vitality and the social function scales. CONCLUSIONS: Mandibular overdentures retained by two implants provide elderly patients with better OHQOL. General health-related quality of life improved in the implant group. PMID- 12752542 TI - Cost-effectiveness of different caries preventive measures in a high-risk population of Swedish adolescents. AB - OBJECTIVES: A total of 3373 12-year-olds agreed to participate in an intervention study evaluating different caries preventive measures. The study, titled 'Evaluation of caries preventive measures', was performed between 1995 and 1999 at 26 dental health clinics throughout Sweden. At the start of the study, the subjects were classified as individuals at high or low risk of developing caries. The high-risk group consisted of 1165 subjects. The children in the high-risk group were randomly assigned to one of four preventive programs. The programs represent a step-wise increase in fluoride content, contact with dental personnel and cost. The aim of the present cost-effectiveness analysis (CEA) study performed from a societal perspective is to compare costs and consequences of caries preventive programs in a caries high-risk population. By 'costs' is meant both treatment costs and costs contributed by the patient and the patient's family. Costs contributed by patients and their families consist of out-of-pocket expenses, transportation costs, and time. Conclusions are that it is important to consider the perspective from which a study is carried out. Costs contributed by the patient and the patient's family have a high impact on total costs for children and younger adolescents but decrease with time as the adolescents get older. The present study shows an incremental cost-effectiveness of 2043 SEK (8.54 SEK = 1 US dollar, December 1999) per averted decayed enamel and dentine missing and filled surface (DeMFS), of which treatment costs represent 1337 SEK using the unit cost for a nurse. This means a yearly cost of approximately 334 SEK. PMID- 12752543 TI - Caries incidence in permanent first molars after discontinuation of a school based chlorhexidine-thymol varnish program. AB - OBJECTIVES: To determine whether the cessation for 3 years of a 24-month program of chlorhexidine-thymol varnish applications would affect caries incidence in the first permanent molars of a population of schoolchildren of middle and lower middle socioeconomic level. METHODS: Two groups of 6-7-year-old schoolchildren, randomized by school-class, were followed up in a clinical trial. One group received applications of chlorhexidine-thymol varnish every 3 months and the other group acted as controls. The program ended after 24 months and its effects were evaluated. Three years later, 55.5% of the schoolchildren were re-examined and the caries increments were documented. RESULTS: At the end of the 24-month program, the treated children had significantly fewer decayed and filled surfaces in permanent molars (lower DFS index) versus the controls. At 3 years after the discontinuation of the program, this difference had disappeared; there were no differences in the incidence of decayed, missing and filled surfaces (DMFS) index in permanent molars between the treated children and the controls. CONCLUSION: The cessation for 3 years of a 3-month program of chlorhexidine-thymol varnish applications resulted in a nonsignificant increase in the prevalence of dental caries in the permanent first molar. PMID- 12752544 TI - Fluoride intake by Brazilian children from two communities with fluoridated water. AB - OBJECTIVES: To determine total daily fluoride intake by young children from two communities in a developing country. METHODS: Fluoride intake from diet and dentifrice was determined for 71 Brazilian children, aged 19-38 months, living in two communities with fluoridated water (0.6-0.8 ppm). The children from Piracicaba attended a full-time day care centre but those from Ibia did not. Fluoride ingested during tooth brushing was determined, and 'duplicate-plate' samples of all foods and beverages ingested during a 2-day period were collected from which fluoride for analysis was extracted by hexamethyldisiloxane (HMDS) microdiffusion. The amount of fluoride ingested (mg F/kg body weight/day) from the diet and dentifrice and the combined fluoride intake were calculated. A limit of 0.05-0.07 mg F/kg body weight/day was considered as the safe threshold for fluoride exposure. RESULTS: The children from Ibia had lower amounts of fluoride in their diets than those from Piracicaba (P < 0.05); no differences in the amount of fluoride ingested from dentifrice were found between the communities. In both communities, the daily fluoride intake from dentifrice was higher than that from the diet (P < 0.05). Most of the children from both communities were exposed to a combined dose (diet + dentifrice) of fluoride above the risk threshold for dental fluorosis. CONCLUSION: The data suggest that fluoride intake from diet depends on living conditions, and measures should be implemented to reduce the fluoride intake of these Brazilian children. PMID- 12752545 TI - The self-assessed oral health status of individuals from White, Indian, Chinese and Black Caribbean communities in South-east England. AB - OBJECTIVES: To determine the level of self-assessed oral symptoms and the impact of such symptoms among individuals from four ethnic groups resident in South-east England and the relationship between self-assessed oral health status, age, gender, employment status, educational level and ethnicity. METHOD: Cross sectional survey of a convenience sample of 366 individuals drawn from four ethnic groups. MEASURES: Subjective Oral Health Status Indicators (SOHSI). PARTICIPANTS: Individuals were recruited through community groups. All participants self-classified their ethnicity. Only completed questionnaires from participants categorising themselves as White, Black Caribbean, Chinese or Indian were included in the data analysis. FINDINGS: Univariate statistical analysis revealed significant differences between ethnic groups in all but one of the SOHSI scales. Age and ethnicity (in particular membership of the Chinese community) emerged as significant predictors of SOHSI scale scores. CONCLUSIONS: Within the limitations imposed by convenience sampling, it has been found that differences exist among four ethnic groups in the UK in their reporting of self assessed oral health status. Ethnicity and age, in particular, predict the reporting of self-assessed oral symptoms and the impact of such symptoms. PMID- 12752546 TI - Reporting on adverse reactions to dental materials--intraoral observations at a clinical follow-up. AB - OBJECTIVES: A national reporting system designed to monitor adverse reactions to dental materials was established in Norway in 1993. The activities have also included clinical examination of patients with suspected reactions to dental materials. The ongoing activities are coordinated by the Dental Biomaterials Adverse Reaction Unit at the University of Bergen. The reporting procedure is based on voluntary spontaneous reporting by dentists and physicians. The reports could be based on subjective symptoms or objective findings, or both. The aim of the present study was to compare reported objective intraoral findings with those found during examination at the unit. METHODS: Reported reactions were compared with clinical findings obtained following dental and medical examination at the unit. From 1993 to 1999, a total of 899 reports were received while 253 patients were referred and examined at the unit. RESULTS: The reports on patients who were examined at the unit involved mainly reactions related to amalgam fillings (84%), metals in fixed dentures (11%), resin-based materials and cements (4%), materials used in removable dentures (2%), and endodontic materials (2%). Edema, lichenoid reactions, ulcers/vesicles, erythema, and atrophy were found in 80 patients during the examination at the unit. For 35 of these patients, the intraoral findings at the unit were also given in the reports. For another 45 patients, objective intraoral signs of reactions were found upon examination at the unit, but these findings had not been reported. CONCLUSION: A spontaneous reporting system is a cost-effective method for monitoring intraoral reactions associated with dental materials. Considering the increasing number and complexity of these materials, there appears to be a need for continuous validation of reports by a speciality unit. In order to receive more accurate information about the adverse reactions, it would be advisable that the reporting forms include more detailed guidance regarding signs of reactions that practitioners should be on the look out for and consider. PMID- 12752547 TI - A comparison of dental health care attitudes in the Netherlands in 1985, 1995, and 2001. AB - OBJECTIVES: The aim of the present study was to re-establish and update empirical data obtained in 1985 and 1995 using the dental attitudes questionnaire (DAQ). In 1984, this instrument for assessing oral health care attitudes was presented by Stockwell and Banting. The DAQ has six content scales: cynicism, health concern, motivation, oral function, social aesthetic and susceptibility, and two validity scales, halo and infrequency. METHODS: The DAQ was completed in a similar setting using similar subjects in 1985, 1995, and 2001. Subjects were first-year psychology students who participated for course credit. RESULTS: A high level of agreement among respondents was found in their ranking of attitudinal factors. The mean scores on most subscales were lower in 1995 and 2001 than in 1985. In 2001, respondents scored higher on the subscale 'motivation', and lower on the subscales 'cynicism' and 'social aesthetic' than subjects in 1995. CONCLUSION: Some changes have occurred in dental health care attitudes as measured by the DAQ during the intervals between the three studies. Subjects in 2001 showed less suspicion regarding the motives of dental health care professionals and downplayed more need for regular dental checkups and oral hygiene than subjects in former years. The psychometric qualities of the DAQ were considered to be satisfactory. PMID- 12752548 TI - Caries and micronutrient intake among urban South African children: a cohort study. AB - OBJECTIVE: To report on associations between caries and micronutrient intake among a 'true cohort' group of 5-year-old urban black South African children. METHODS: The study sample was a true cohort that had nutrition as well as dental information for 1991 and 1995 (n = 259). Micronutrient intake and dental caries associations were examined with SAS using the linear logistic analysis and a critical level of statistical significance of P < 0.05. RESULTS: Of the 21 micronutrients investigated, vitamin B12, riboflavin, magnesium and biotin were individually significantly associated with caries incidence, but taken together only magnesium was significant. CONCLUSIONS: The association of micronutrient intake with caries was weak, isolated and not clinically relevant. Micronutrient intake was not an indicator of the presence or absence of caries among the children studied. PMID- 12752549 TI - Fluoride intake from foods, beverages and dentifrice by children in Mexico. AB - OBJECTIVES: This investigation monitored fluoride ingestion in Mexican children who were susceptible to developing dental fluorosis. METHODS: Total fluoride intake, from dietary and toothpaste samples, was determined in 15-36-month-old children from Mexico City and Veracruz, Mexico. A duplicate plate technique was used. Plasma fluoride levels were also determined in this group of children. RESULTS: The children ingested a mean (+/-SD) of 0.20 +/- 0.08 and 0.18 +/- 0.07 mg fluoride/kg/day, in Mexico City and Veracruz, respectively. There was no statistically significant difference between the fluoride ingested by children in the two cities (P > 0.9). Plasma from children in Mexico City and Veracruz contained a mean of 1.30 +/- 0.81 and 0.87 +/- 0.72 micro mol fluoride/l, respectively, and these values were not significantly different. CONCLUSIONS: Our results lead to the conclusion that mean fluoride ingested from the combination of foods and beverages was within the proposed safe threshold for fluoride intake of 0.05-0.07 mg F/kg/day. Most of the fluoride intake by these children was derived from the ingestion of fluoridated toothpaste. When all sources of ingested fluoride were added and total fluoride intake was calculated, the children, both in Mexico City and Veracruz, were ingesting amounts of fluoride well above the upper limits of the proposed safe threshold for fluoride intake. PMID- 12752550 TI - Classification of dental caries patterns in the primary dentition: a multidimensional scaling analysis. AB - OBJECTIVE: Clinical patterns of early childhood caries (ECC) encompassing specific teeth or surfaces have been previously proposed on an a priori basis and have been used as case definitions. The underlying assumption is that the patterns result from different host response and environmental conditions. Identifying caries patterns has utility in refining case definitions of ECC. Well defined caries patterns should enhance the ability of an analysis to identify meaningful associations between suspected risk factors and ECC. The purpose of this project was to identify patterns of caries in the dentition of preschool children using multidimensional scaling without a priori pattern delineation. METHODS: Between February 1994 and September 1995, five examiners visually examined 5171 Arizona preschool children aged 5-59 months old. Multidimensional scaling (MDS) was used as a classification/taxonomy technique to identify any underlying structure of the caries data. MDS provided a classification scheme for individual tooth surfaces based on the dissimilarity measures of squared Euclidean distance and of variance using an alscal analysis. Both two- and three dimensional solutions were pursued; s-stress, stress, R-square and residual patterns were assessed in determining the best dimensional model, with the resulting quadrant positions of the tooth surfaces suggesting potential caries patterns. RESULTS: All models demonstrated excellent fit. Two- and three dimensional solutions suggested four caries patterns: (i) any maxillary incisor surfaces, (ii) first molar occlusal surfaces, (iii) second molar pit and fissure surfaces, and (iv) any smooth surfaces, excluding the maxillary incisor surfaces. CONCLUSIONS: This is the first delineation of primary dentition caries patterns produced by a classification analysis without a priori pattern definitions. The identified caries patterns may arise from specific risk factors and/or be a function of the timing of various risk factor exposures. Use of these patterns as case definitions should enhance the ability to identify associations between suspected risk factors and ECC. PMID- 12752552 TI - Alveolar collapse and closure of airways: regular effects of anaesthesia. AB - Oxygenation is impaired in almost all subjects during anaesthesia, and hypoxaemia for shorter or longer periods is a common finding. Moreover, postoperative lung complications occur in 3-4% after elective surgery, and up to 20% in emergency operations. Rapid collapse of alveoli on induction of anaesthesia and more widespread closure of airways seem to explain the oxygenation impairment and may also contribute to postoperative pulmonary infection, Causative mechanisms to atelectasis and airway closure seem to be loss of respiratory muscle tone and gas resorption. Avoiding high inspired oxygen fractions during both induction and maintenance of anaesthesia prevents or reduces atelectasis, while intermittent 'vital capacity' manoeuvres, recruit atelectatic lung regions. PMID- 12752553 TI - Variations in percentage of body fat within different BMI groups in young, middle aged and old women. AB - OBJECTIVES: The objective of the present study was to characterize age-dependent variations in percentage of body fat within different body mass index (BMI) classes in healthy Danish women. DESIGN: Cross-sectional analysis. SETTINGS: The study was done at the Center for Clinical and Basic Research, Ballerup, Denmark. SUBJECTS: Four hundred and four healthy women aged 18-75 years were included in the present study. MEASUREMENTS: Fat tissue mass was estimated using dual-energy X-ray absorptiometry. Menopausal status and physical characteristics were also registered. RESULTS: Mean values of percentage of body fat calculated in the normal and overweight BMI groups were higher in middle-aged and old women compared with young women. No significant differences were seen in the underweight and obese BMI groups. The cut-off levels between normal and overweight and between overweight and obesity were 35-43% and 40-50%, respectively. CONCLUSION: The percentage of body fat is dependent on both age and menopausal status within each of the following BMI classes: from 20 to 25 and from 25 to 30, further emphasizing that BMI has limitations when used generally as an indicator of body fatness, and argues for defining BMI cut-off values age specifically. PMID- 12752554 TI - Relation between the elastic properties and intima-media thickness of the common carotid artery. AB - The purpose of the study was to describe the relation between wall elasticity and intima-media thickness in the human carotid artery prior to the development of atherosclerotic plaques. Fifty-eight apparently healthy men, aged 42-65 years (mean 55 years), without symptoms of cardiovascular disease were studied. Thickness and elastic properties of the common carotid artery wall were assessed using ultrasonography and non-invasive arterial pressure measurements. The relation between the calculated intima-media area and the pressure strain elastic modulus was positive and statistically significant on the right but not on the left side. No statistically significant relations were found between the calculated intima-media area and the stiffness or between the intima-media thickness and the elastic modulus or stiffness on either side. Thus, the relations between the common carotid artery intima-media thickness/calculated intima-media area and the common carotid artery elastic modulus/stiffness are weak. In regions without atherosclerotic plaques, the elastic properties of the human carotid artery wall do not seem to be influenced by the wall thickness in an important way. PMID- 12752555 TI - Nasal neutrophil activity and mucinous secretory responsiveness in COPD. AB - Patients with chronic obstructive pulmonary disease (COPD) frequently report nasal symptoms. In the present study, we have examined whether or not COPD is associated with any nasal inflammation. Plasma exudation evoked by histamine challenges has been employed to improve the recovery of inflammatory indices in nasal lavage fluids. In 23 COPD-patients and 26 healthy subjects, all without history or signs of allergic rhinitis, nasal polyposis, or chronic rhinosinusitis, nasal saline-lavages were performed with and without histamine. alpha2-Macroglobulin, fucose, eosinophil cationic protein (ECP) and myeloperoxidase (MPO) were determined as indices of plasma exudation, mucinous secretion, eosinophil activity and neutrophil activity, respectively. The difference in MPO-levels between the histamine and the saline lavage was greater in COPD patients compared with healthy subjects (P<0.05). Also, COPD patients reporting nasal symptoms presented an increase in MPO at histamine challenge (P<0.05, cf. saline) and greater differences in MPO and fucose, respectively, between the histamine and the saline lavage (P<0.05, cf. patients without symptoms). We conclude that COPD is not associated with any marked nasal inflammation. However, our observation on increased MPO-levels at histamine challenge suggests some degree of increased neutrophil activity in this condition. Furthermore, when associated with nasal symptoms, COPD may be associated with an increased nasal secretory responsiveness. PMID- 12752556 TI - Oxygen tension under hyperbaric conditions in healthy pig brain. AB - OBJECTIVE: To investigate the effect of hyperbaric conditions on brain oxygenation, intracranial pressure and brain glucose/lactate levels in healthy non-brain-traumatized animals. DESIGN AND SETTING: Prospective animal study in a hyperbaric chamber. SUBJECTS: Twelve adult Landrace/Yorkshire pigs. INTERVENTIONS: The animals were normoventilated in a pressure-controlled mode according to the open lung concept first at normobaric pressures (FiO2 of 0.4 and 1.0) and subsequently in the hyperbaric chamber at 1.9 and 2.8 bar (both at an FiO2 of 1.0). Under these conditions brain oxygen tension and intracranial pressure were recorded and brain glucose/lactate levels were measured by microdialysis. RESULTS: At normobaric conditions, increasing the FiO2 from 0.4 (baseline) to 1.0 resulted in a significant increase in brain oxygen tension from 33 +/- 14 to 63 +/- 28 mmHg (P<0.05). Compared with baseline, both hyperbaric conditions (at an FiO2 of 1.0) led to a significant increase in brain oxygen tension to 151 +/- 65 mmHg (P<0.001) at 1.9 bar and to 294 +/- 134 mmHg (P<0.001) at 2.8 bar. CONCLUSIONS: If there is a need for increased oxygenation in the brain, then one way to achieve this is to apply hyperbaric conditions at 100% oxygen. Compared with an atmospheric pressure with a FiO2 of 0.4, a nine-fold increase (900%) in PbrO2 values can be reached by increasing the FiO2 to 1.0 and the pressure to 2.8 bar. In this study, hyperbaric oxygen pressure in the brain did not lead to changes in intracranial pressure or in brain glucose/lactate levels. PMID- 12752557 TI - Hyperbaric hyperoxia induces a neuromuscular hyperexcitability: assessment of a reduced response in elite oxygen divers. AB - We compared the changes in compound muscle mass action potential (M-wave) recorded in vastus lateralis in response to hyperbaric hyperoxia (HBO) in nine combat divers who dived daily while breathing 100% O2 or O2-enriched mixture (O2 divers) to those measured in eight recreational divers who dived occasionally using compressed air/21% O2 (air divers). The O2 divers completed a 6-h HBO exposure in which the inspired oxygen pressure (PiO2) varied from 1.15 to 2.7 absolute atmospheres (ATA), PiO2 being maintained at 1.15 ATA throughout the first 2-h period, whereas the air divers only completed a 2-h HBO exposure with PiO2 constant at 1.15 ATA. Before HBO exposure, there were no intergroup differences between baseline M-wave characteristics (amplitude and duration), but the conduction time was significantly shorter in O2 divers compared with air divers. After 90 min of HBO (1.15 ATA) the air divers demonstrated neuromuscular hyperexcitability, as evidenced by an increased M-wave amplitude (13%, P<0.01 versus baseline), shortened M-wave duration (5%, P<0.05 versus baseline), and reduced conduction time (5%, P<0.01 versus baseline). In O2 divers, similar HBO induced M-wave changes were only observed when PiO2 was greater than 1.50 ATA. We conclude that HBO elicites neuromuscular hyperexcitability, attenuated in elite O2 divers. PMID- 12752558 TI - Effects of experimental changes in nasal airway pressure on mucosal output of plasma. AB - Microvascular extravasation, lamina propria flooding and luminal entry of plasma are key features of airway inflammation. We have suggested that the extravasated plasma moves across the epithelial lining along hydrostatic pressure gradients. The present study, involving healthy subjects, tests this hypothesis by examining effects of experimentally applied negative and positive luminal pressures on nasal output of plasma at baseline and at histamine-induced plasma exudation. The negative (-10 cmH2O) and positive (10 cmH2O) pressures were applied for 10 min after nasal spray administrations of diluent (saline) and histamine (0.5 mg). The mucosa was then lavaged and the lavage fluid levels of alpha2-macroglobulin were measured as index of plasma exudation. Nasal administrations of diluent and histamine (0.5 mg) were also carried out without any pressure applications. Histamine produced significant mucosal exudation of plasma. The negative luminal pressure augmented this response significantly as well as the baseline appearance of alpha2-macroglobulin in mucosal surface liquids. We conclude that extravasated plasma may be moved across the epithelium by a hydrostatic pressure-operated epithelial mechanism. PMID- 12752559 TI - Effect of non-hypotensive haemorrhage on plasma catecholamine levels and cardiovascular variability in man. AB - BACKGROUND: It is well known from animal research that non-hypotensive haemorrhage produces sympathoexcitatory responses assessable by both the rise in plasma catecholamine levels and the shift of autonomic influences on the heart to more sympathetic and less parasympathetic control. Data in humans are restricted. METHODS: Heart rate variability (HRV), systolic blood pressure (FINAPRES) variability (BPV), and catecholamine plasma levels were measured before and after haemorrhage in 30 healthy blood donors and compared with those from 10 control subjects without blood loss. Spectral power of HRV and BPV in very low (0.02-0.06 Hz), low (0.07-0.14 Hz), and high (0.15-0.40 Hz) frequency bands were calculated by Fourier analysis. Catecholamine plasma levels were assayed by dual column reverse-phased high-performance liquid chromatography (HPLC). RESULTS: Haemorrhage of 5.6 +/- 1.2 ml kg-1 body weight increased plasma norepinephrine levels (215 +/- 92 pg ml-1 versus 254 +/- 95 pg ml-1; P = 0.002), increased BPV in the low frequency band (Mayer waves; 1.8 +/- 1.0 ln [mmHg(2)] versus 2.0 +/- 0.9 ln [mmHg(2)]; P = 0.021), and decreased the vagally transmitted high frequency HRV (6.9 +/- 1.1 ln [MI(2)] versus 6.5+/-1.2 ln [MI(2)]; P<0.0001), but did not induce significant changes in heart rate (66 +/- 11 bpm versus 67 +/- 11 bpm; P = 0.79) and arterial blood pressure (mean values: 84 +/- 13 mmHg versus 87 +/- 13 mmHg; P = 0.12). CONCLUSIONS: As suggested by plasma norepinephrine levels, systolic BPV and HRV, non-hypotensive haemorrhage produces sympathoexcitatory responses as well as vagal withdrawal of heart rate control in humans. PMID- 12752560 TI - Reference values for the physical work capacity on a bicycle ergometer for men -- a comparison with a previous study on women. AB - The aim of the study was to collect new reference values for the clinical ramp exercise test on bicycle, because in our experience, the commonly used values were too low. A group of healthy men (n = 81, 20-80 years) was randomly selected from the local municipal register to achieve an even distribution in age. Data were compared with those obtained in a similar, previous study on women (n = 87). The subjects were encouraged to cycle until exhaustion (19 on the Borg scale) when maximal load, heart rate and systolic blood pressure were recorded. Maximal load (W(max)) was related to age (years) and height (m) using a non-linear function: W(max) = (244.6 x height - 92.1)/[1 + exp[0.038 x (age - 77.3)]]. Maximal heart rate (HRmax) was described by a similar function: HRmax = 203.7/[1 + exp[0.033 x (age - 104.3)]]. The maximal systolic blood pressure (BP(max)) was described by a linear function based on age: BP(max) = 0.505 x age + 192. Similar functions for the women are also given. It is suggested that 80-120% of the predicted maximal load can be taken as a reference interval for both men and women and similarly 90-110% of the maximal heart rate. In this study, 84% of the men reached a maximal load within the reference interval and 93% maximal heart rate within the reference interval. The reported values for maximal load were 104 132% of the reference values published by others. PMID- 12752561 TI - Postural sway in normal subjects aged 20-70 years. AB - So far, no population-based reference material of postural sway has been presented in the literature. This study evaluated postural control by measuring posturographic parameters, with the aim of establishing useful standards with regard to gender, age, body weight, cigarette smoking, alcohol consumption and articular hypermobility. This was performed in an age-stratified random sample of 195 subjects from the Copenhagen City Heart Study, of whom, 133 agreed to participate. Measurement of postural sway was performed the Balance Master Pro(R). Outcome parameters were average angular velocity, target sway area, per cent maximum stability and per cent ankle strategy. Using SAS Proc Mixed, backward stepwise elimination was performed and 95% prediction intervals were obtained. Sway increased with increasing difficulty of the test (P<0.001) and with increasing age (P<0.001). No differences were found between the sexes. Ankle strategy did not change with age, but decreased with increasing body weight, and increased in subjects reporting a consumption of modest amounts of alcohol (P<0.001), or with the finding of increased hypermobility (P = 0.006). The present material may be used as a reference in further studies of postural control. PMID- 12752562 TI - It's only teeth - are there limits to genetic testing? AB - Dental genetic disorders can cause severe social and psychological effects in affected individuals. The cost of treatment can be considerable, not only in financial terms but also in time spent during treatment. In theory it is, or will soon be, possible to use advances in molecular genetics for pre-natal testing, for selection of embryos using in vitro fertilization techniques, and for gene therapy. The questions we pose are whether these approaches are appropriate. We hope that this review will stimulate debate on these issues. PMID- 12752563 TI - Vascular malformations: localized defects in vascular morphogenesis. AB - Vascular anomalies are localized defects of the vasculature, and usually affect a limited number of vessels in a restricted area of the body. They are subdivided into vascular malformations and vascular tumours. Most are sporadic, but Mendelian inheritance is observed in some families. By genetic analysis, several causative genes have been identified during the last 10 years. This has shed light into the pathophysiological pathways involved. Interestingly, in most cases, the primary defect seems to affect the characteristics of endothelial cells. Only mutations in the glomulin gene, responsible for hereditary glomuvenous malformations, are thought to directly affect vascular smooth-muscle cells. PMID- 12752567 TI - Analysis of marker or complex chromosomal rearrangements present in pre- and post natal karyotypes utilizing a combination of G-banding, spectral karyotyping and fluorescence in situ hybridization. AB - The significance of complex chromosomal rearrangements presents a diagnostic dilemma. In the past, the use of G-banding coupled with fluorescence in situ hybridization (FISH) has been the standard approach. The recent development of spectral karyotyping (SKY) and multicolor FISH (M-FISH) has resulted in an increased accuracy of identification of marker or other complex chromosomal rearrangements. However, owing to the additional cost and time associated with SKY or M-FISH, and the restricted availability of such imaging facilities in many centers, it is not feasible to perform these procedures routinely on every sample. In addition, the identification of an aberration by SKY or M-FISH will often require confirmation by FISH. A practical approach is needed to take advantage of the complementary strengths of each method. In our center we utilize an algorithm that dictates the use of routine G-banding for the initial preliminary evaluation of a patient, followed by SKY characterization if marker chromosomes or complex translocations are detected by the G-banding analysis. According to this algorithm, FISH is used to verify the results once the origin of the abnormal chromosome has been determined by SKY. To demonstrate the effectiveness of this algorithm, we have analyzed both amniocyte and lymphocyte slides, using a combination of G-banding, SKY, and FISH. Our results confirm that an algorithm which selectively uses SKY or M-FISH will provide an efficient and improved method for pre- and post-natal chromosomal analysis. PMID- 12752568 TI - Pulmonary involvement in type 1 Gaucher disease: functional and exercise findings in patients with and without clinical interstitial lung disease. AB - Pulmonary disease is a well-known complication of Type 1 Gaucher disease (GD), although its incidence is not well established and its severity varies. The purpose of this study was to determine the frequency and extent of pulmonary involvement in patients with GD. Pulmonary involvement was assessed by history, physical examination and chest radiograph in 150 consecutive patients with Type 1 GD presenting at a specialized center for genetic diseases. Five patients were noted to have clinical evidence of pulmonary involvement. Full pulmonary function tests were performed in these five patients and in an additional 13 patients randomly selected from the remaining 145. Many of the 18 patients also underwent radionuclide body imaging with 67 Gallium citrate and 111Indium-tagged leucocyte scans, as well as incremental cardiorespiratory exercise tests. Lung biopsies were available in two patients with lung disease, and a second examination of lung tissue was performed in one of these two patients post-mortem. Clinical lung disease was detected in five patients. All five had dyspnea, diffuse infiltrates, restrictive impairment and low single breath CO diffusing capacity (DLCOSB). Two of these patients underwent exercise testing and showed abnormalities consistent with lung disease (ventilatory limitation, excessive ventilation and increased dead space) as well as decreased VO2 max. and anaerobic threshold (AT). In contrast, in the other 13 patients, physical examination, chest radiographs and pulmonary function were normal (except for a low DLCOSB in one patient). Responses on exercise testing (performed in six of the 13 patients) were consistent with a circulatory impairment (decreased VO2 max. and AT). Our study found that <5% of patients with Type 1 GD have clinical interstitial lung disease. In addition, we found that some patients, without evident lung involvement, may experience limitations in physical exertion and are easily fatigued; this is attributable to impaired circulation. PMID- 12752569 TI - APOA5-1131T>C polymorphism is associated with triglyceride levels in Chinese men. AB - A new apolipoprotein gene, APOA5, was recently discovered near the APOA1/C3/A4 gene cluster. Transgenic mice overexpressing the homologous gene, apoa5, showed reduced plasma triglyceride levels, while knockout mice had greatly increased triglycerides, suggesting that human genetic variants affecting expression of the protein product, APOAV, might affect triglyceride levels. Polymorphisms in the APOA5 gene were indeed found to be associated with triglyceride levels in men, though not in women. We sought to confirm the association of the APOA5-1131T>C polymorphism with triglyceride levels in 167 Chinese men chosen for having either high (>/=1.7 mm, n = 82) or low (T (5.7%), G542X (3.9%), N1303K (2.6%), 2789+5G>A (1.7%), 2183AA>G (1.4%), E822X (1.4%), R1158X (1%)], 12 showed frequencies between 0.5% and 1%, while the remaining (60) were very rare (1 to 3 alleles). Denaturing gradient gel electrophoresis (DGGE) screening of 12 exons (3, 4, 7, 10, 11, 13, 14b, 16, 17b 20 and 21) detected 85.5% of CF alleles. Haplotypes for eight diallelic and three microsatellite markers have been characterized for the common, a few rare and novel Greek mutations. Results of 165 prenatal diagnoses (including 49 due to bowel hyperechogenicity), testing a total of 41 different parental genotypes, are reported. One hundred and sixteen prenatal tests resulted in 22 affected, 59 heterozygous, 34 normal fetuses and one incomplete diagnosis. Of the 49 echogenic bowel fetuses, 3 were heterozygotes. Carrier screening was initiated, with emphasis on individuals and couples in high-risk groups - with a family history of CF, one partner with CF, and couples with male infertility seeking in vitro fertilization (IVF). Mutation analysis on 672 individuals (120 couples, 91 unaffected CF siblings, 283 CF family relatives and 58 general population subjects), identified a total of 176 heterozygotes and 7 couples where both partners were CF heterozygotes. Prenatal diagnosis was performed in 4 cases and 3 were counseled on the availability of a prenatal test. PMID- 12752574 TI - Five new subjects with ring chromosome 22. AB - Ring chromosome 22, a rare cytogenetic finding, was first described by Weleber et al. in 1968. Since then approximately 50 patients have been reported in the medical literature. We describe five previously unreported subjects with ring chromosome 22 syndrome, summarize the clinical findings of reported patients from the literature and discuss the involvement of the ring chromosome and clinical outcome. Our subjects demonstrated the prominent features of this syndrome including mental retardation, hypotonia, motor delay, lack of speech, full eyebrows, and large ears. In addition, two of our subjects had central nervous system malformations and regression. The lack of consistent physical abnormalities in our subjects further supports no consistent phenotype manifestations in this cytogenetic syndrome. The variable clinical manifestations seen in ring chromosome 22 subjects may be associated with loss of chromosome 22 sequences near the telomere or attributed to the genetic background of each subject. Similarly, recessive alleles unmasked by the deletion could also contribute to the phenotype. PMID- 12752575 TI - Triple A syndrome: genotype-phenotype assessment. AB - The triple A or Allgrove syndrome is an autosomal-recessive disease (MIM*231550) characterized by the triad of achalasia, alacrima and adrenocorticotropic hormone (ACTH)-resistant adrenal insufficiency. Associated features of the syndrome are neurological and dermatological abnormalities. Until the discovery of the AAAS gene as the responsible gene in triple A syndrome, the diagnosis was based on characteristic clinical features. Here we present the clinical and molecular genetic data which demonstrated the marked phenotypic variability in three unrelated patients with triple A syndrome. The final diagnosis of triple A syndrome was confirmed by molecular analysis. In one patient with isolated achalasia, the diagnosis of triple A syndrome could only be made on the basis of the molecular genetic analysis of the AAAS gene. We therefore suggest that the diagnosis of triple A syndrome should be considered in patients who exhibit only one or two of the main symptoms (i.e. alacrima, achalasia or adrenal insufficiency). These patients require careful neurological investigation, and mutation analysis of the AAAS gene should be performed. PMID- 12752576 TI - Congenital universal hypertrichosis with deafness and dental anomalies inherited as an X-linked trait. AB - We report a large Mexican kindred with a variant form of congenital universal hypertrichosis that is inherited in an apparent X-linked recessive manner. In addition to the generalized hypertrichosis, the affected individuals have dental malformations and deafness. Males are more severely affected than females who exhibit only mild hypertrichosis, but not deafness or dental anomalies. Haplotype analysis in this pedigree revealed linkage to a 13-cM region on chromosome Xq24 q27.1 between markers GATA198A10 and DXS8106. Localization of the gene underlying this form of hypertrichosis is the initial step in identifying genes on the X chromosome that are involved in the control of hair growth and development. PMID- 12752577 TI - Exclusion of PTPN11 mutations in Costello syndrome: further evidence for distinct genetic etiologies for Noonan, cardio-facio-cutaneous and Costello syndromes. AB - Costello syndrome (CS) is a rare, multiple congenital anomaly syndrome with characteristic dysmorphic features, cardiac anomalies and a tendency to develop certain cancers. Phenotypically there is some overlap with other genetic disorders, notably cardio-facio-cutaneous (CFC) syndrome and Noonan syndrome (NS), suggesting that these syndromes may be allelic. We recently identified PTPN11, which encodes the non-receptor protein tyrosine phosphatase, SHP-2, as a major NS disease gene. In this report, we screened a cohort of 27 patients, with the clinical diagnosis of CS, for PTPN11 mutations using denaturing high performance liquid chromatography analysis. No mutations of the PTPN11 gene were found in the CS patients. Common polymorphisms in introns 6 and 7 and exon 8 were identified in four individuals. With our previous exclusion of PTPN11 mutations in CFC syndrome, these data suggest distinct genetic etiologies for Noonan, CFC and Costello syndromes. PMID- 12752578 TI - Discordant clinical manifestations in monozygotic twins with the identical mutation in the TSC2 gene. PMID- 12752579 TI - Do neurotrophins play a role in allergic diseases and asthma? PMID- 12752580 TI - What can be learned about the fibrosis of asthma from rodent models? PMID- 12752581 TI - New insights into the role of the mast cell in asthma. PMID- 12752582 TI - CD4+ T cell epitope identification: applications to allergy. PMID- 12752583 TI - Neonatal interleukin-12 capacity is associated with variations in allergen specific immune responses in the neonatal and postnatal periods. AB - BACKGROUND AND OBJECTIVES: A reduced capacity of antigen presenting cells (APC) to provide pro-T helper 1 (Th1) signals, such as IL-12, to T cells during early life may be implicated in the development of T helper 2 (Th2)-mediated allergic disease. In this study we examined the relationships between the capacity for IL 12 responses in the neonatal period and atopic risk (family allergy), in vitro T cell responses to allergens, and the subsequent development of allergic disease at 6 years. METHODS: The capacity of circulating neonatal (and maternal) APC to produce IL-12 p70 in response to LPS (and IFN-gamma) stimulation was assessed in a group of 60 children with previously well-characterized immune responses to allergens and atopic outcomes. The IL-12 responses were compared with allergen induced lymphoproliferation (to house dust mite (HDM) ovalbumin (OVA), cat and beta-lactoglobulin (BLG)) and IL-13 and IFN-gamma cytokine responses (to OVA, HDM and phytohaemaglutinin (PHA)) in the neonatal and postnatal periods. IL-12 responses were also compared according to atopic risk and atopic outcomes (doctor diagnosed asthma, eczema, food allergies and sensitization as evidenced by skin prick testing) at 6 years clinical follow-up. RESULTS: Maternal peripheral blood mononuclear cells (PBMC) synthesized significantly greater amounts of IL-12 than neonatal PBMC, though within maternal-infant pairs IL-12 responses were significantly correlated (r = 0.4, P = 0.019). Moreover, neonatal IL-12 responses were positively correlated with neonatal allergen proliferation for HDM (r = 0.6, P < 0.0001), OVA (r = 0.55, P < 0.0001), cat (r = 0.5, P = 0.003) and BLG (r = 0.55, P = 0.001), but negatively correlated with neonatal IL-13 responses to both allergens tested (HDM: r = - 0.4, P = 0.03 and OVA: r = - 0.5, P = 0.001). Both neonatal and maternal IL-12 responses were positively correlated with postnatal IFN-gamma responses to HDM at 12, 18 and 24 months of age (responses after age of 2 years were not assessed). There was no relationship between atopic risk and IL 12 capacity in the neonatal period, but there was a (non-significant) trend for neonatal IL-12 responses to be lower in the high-risk children who developed clinical allergy at 6 years (compared with the low risk group) although the number in this analysis was small. CONCLUSIONS: Reduced APC IL-12 production in the perinatal period was associated with reduced T cell activation (lymphoproliferation), stronger neonatal Th2 responses, and weaker Th1 responses to allergen in the postnatal period. This supports the notion that variations in APC function in early life may contribute to altered allergen-specific cytokine responses associated with later allergy. PMID- 12752584 TI - Characterization of wheezing phenotypes in the first 10 years of life. AB - BACKGROUND: Childhood wheezing illnesses are characterized into different phenotypes. However, severity of the disease associated with these phenotypes has not been extensively studied. OBJECTIVES: To determine characteristics of childhood wheezing phenotypes in the first decade of life using health outcomes plus measurements of atopy, lung function and bronchial hyper-responsiveness. METHODS: A whole population birth cohort (n = 1456) was prospectively studied to examine the natural history of childhood wheezing. Children were seen at 1, 2, 4 and 10 years for questionnaire completion and prospectively collected data used to define wheezing phenotypes. Assessment was made of adverse health outcomes plus spirometry, bronchial hyper-responsiveness, serum IgE measurement at 10 years and skin test sensitization at both 4 and 10 years for wheezing phenotypes. RESULTS: Phenotypic analysis identified that 37% early life wheezers (symptom onset by age 4 years) still wheezed at 10 years. These persistent wheezers showed significantly more physician-diagnosed asthma in early life (P < 0.005 at 2 years) than early transient wheezers (wheezing transiently with onset by age 4 years). Overall they experienced greater multiple hospital admissions (P = 0.024), specialist referral (P = 0.009) and use of inhaled (P < 0.001) and oral steroids (P < 0.001) than early transient wheezers. They also demonstrated enhanced bronchial hyper-responsiveness compared with early transient wheezers (P < 0.001). However, both groups of early life wheezers showed impairment of baseline lung function at 10 years in comparison with non-wheezers: FEV1 (P < 0.029) and FEV1/FVC ratio (P < 0.001) with persistent wheeze and PEF (P = 0.036) with early transient wheeze. Late-onset wheezers (onset from 5 years onwards) had similar BHR to persistent wheezers but maintained normal lung function at age 10 and had lower cumulative prevalence of adverse health outcomes than persistent wheezers. CONCLUSIONS: Persistent wheezing with early childhood onset is associated with substantial morbidity in the first decade of life in association with high levels of atopy, bronchial hyper-responsiveness and impaired lung function at 10 years of age. Late-onset wheezing in the first decade of life could harbour potential for similarly significant disease subsequently. PMID- 12752585 TI - Mucosal and systemic inflammatory changes in allergic rhinitis and asthma: a comparison between upper and lower airways. AB - BACKGROUND: Local airway inflammation and airway remodelling are considered important in the clinical expression of allergic asthma. OBJECTIVE: The aim of this study was to compare airway inflammation and remodelling in nasal and bronchial mucosa of subjects with allergic rhinitis with or without asthma. METHODS: Four experimental groups were formed: allergic asthma and rhinitis (n = 19); allergic rhinitis, no asthma (n = 18); atopic subjects, no asthma, no rhinitis (n = 8) and non-allergic healthy control subjects (n = 16). Blood samples, nasal and bronchial biopsy specimens were collected during stable disease. Immunohistochemistry was performed for eosinophils (MBP), mast cells (CD117) and vascular endothelium (CD31). Epithelial loss, reticular basement membrane (RBM) thickness and subepithelial vascularity was assessed with a computer-assisted image analysis system. RESULTS: In nasal and bronchial mucosa, numbers of eosinophils were significantly higher in rhinitis patients with and without asthma than in asymptomatic atopics (P < 0.05) and controls (P < or = 0.01). In bronchial mucosa, the RBM was significantly thickened in rhinitis patients with and without asthma compared to asymptomatic atopics (P < 0.05) and controls (P < 0.01), while in nasal mucosa no differences were seen. Patients with asthma and rhinitis had increased numbers of blood eosinophils (P = 0.05) and skin test reactivity (P = 0.01) compared to patients with rhinitis only. No significant differences could be found between the investigated groups with respect to serum IL-5 and eotaxin levels, the number of mucosal mast cells and the degree of epithelial loss and subepithelial vascularity. Epithelial desquamation was significantly increased in the bronchial mucosa compared to nasal mucosa, not only in asthmatics (P < 0.001), but also in atopics without asthma and rhinitis (P = 0.02). CONCLUSIONS: This study shows that allergic inflammation, increased basement membrane thickness and epithelial desquamation are present in the lower airways of atopic subjects, even before the onset of clinical symptoms. Despite the presence of inflammatory cells, no structural changes could be assessed in nasal mucosa of allergic patients. PMID- 12752586 TI - Longitudinal decline in pulmonary function in atopic cough and cough variant asthma. AB - OBJECTIVE: Cough variant asthma and atopic cough are different clinical manifestations of eosinophilic airway inflammation presenting with isolated chronic non-productive cough. The aim of this study was to examine the longitudinal change in pulmonary function in cough variant asthma and atopic cough. METHODS: Longitudinal change in FEV1 was prospectively examined in 20 patients with cough variant asthma, 14 patients with atopic cough and 271 asymptomatic healthy subjects. All were lifetime non-smokers. Of the 20 cough variant asthma patients, 13 were taking long-term inhaled corticosteroid therapy (ICS) (beclomethasone dipropionate 615 +/- 58 micro g/day) and the other seven were not. Spirometry was taken at first visit, after cough was almost completely relieved on therapy, and at least once every year for 5 or more years afterwards. RESULTS: The slope of longitudinal change in FEV1 was not significantly different among cough variant asthma patients (- 0.029 +/- 0.007/year), atopic cough patients (- 0.021 +/- 0.022/year) and asymptomatic subjects (- 0.028 +/- 0.002 L/year). In patients with cough variant asthma, the slope in patients not taking inhaled corticosteroids (ICS) was 0.032 +/- 0.007 L/year, which was not significantly different from that in patients taking ICS (- 0.027 +/- 0.010 L/year). CONCLUSION: Pulmonary function decline is not greater in cough variant asthma than atopic cough and the normal population, and long-term ICS has no effect on the decline in cough variant asthma. PMID- 12752587 TI - Increased levels of vascular endothelial growth factor in induced sputum in asthmatic patients. AB - BACKGROUND: Vascular endothelial growth factor (VEGF) is highly expressed in the airway of asthmatic patients. As VEGF increases airway vascular permeability, consequent thickening of the airway wall mucosa may lead to narrowing of the airway lumen. OBJECTIVE: We evaluated the relationship between VEGF levels in induced sputum and eosinophilic inflammatory profiles, and the degree of airway vascular permeability in asthmatic patients and we evaluated the effect of inhaled corticosteroids on VEGF levels in induced sputum. METHODS: Induced sputum specimens were obtained from 28 glucocorticosteroids free asthmatics and 11 healthy control subjects. We examined VEGF levels and airway vascular permeability index in induced sputum. After the initial sputum induction, 21 asthmatics received 8-week inhaled beclomethasone dipropionate (BDP, 800 micro g/day) therapy, then sputum induction was repeated. RESULTS: The VEGF levels in asthmatics were significantly higher than in healthy control subjects (P < 0.0001). The VEGF levels were negatively correlated with forced expiratory volume of 1 s (FEV1, % predicted, r = - 0.68, P < 0.001), the percentage of eosinophils (r = 0.51, P < 0.01) and ECP levels (r = 0.39, P < 0.05). Moreover, the VEGF levels were significantly correlated with airway vascular permeability index (r = 0.61, P < 0.001). After 8-week inhaled BDP therapy, the VEGF levels were significantly decreased compared to pretreatment levels (P < 0.0001) and the VEGF levels were significantly correlated with airway vascular permeability index even in post-treatment asthmatics (r = 0.62, P < 0.01). CONCLUSION: The VEGF levels in induced sputum were increased in asthmatics and its levels were associated with degree of airway narrowing and airway vascular permeability. These findings provide strong evidence that VEGF may play an important role in the pathogenesis of bronchial asthma. PMID- 12752588 TI - Food hypersensitivity among Finnish university students: association with atopic diseases. AB - BACKGROUND: Food hypersensitivity (FH) is commonly suspected, especially among adults with atopic diseases. Symptoms of FH vary from oral allergy syndrome (OAS) to gastrointestinal, respiratory and systemic reactions. More data are needed regarding patient groups at risk for FH, and symptoms and foods responsible for the reactions. METHODS: FH was studied in 286 Finnish university students. Four study groups were selected: subjects (i) with current atopic dermatitis (AD) with or without allergic rhinoconjunctivits (ARC) or asthma (n = 41); (ii) with past AD with or without ARC or asthma (n = 89); (iii) with ARC or asthma (n = 69); (iv) without clinically confirmed atopic disease (n = 87). A thorough clinical examination was performed with a questionnaire specifying adverse events to foods. In addition, IgE specific to five foods, and skin prick tests to four foods were determined. RESULTS: FH was reported by 172 subjects (60.1%), more often by females (66.3%) than by males (47.9%) (P = 0.003). FH was most frequent among subjects with AD, among those with current AD in 73.2%, with past AD in 66.3%, and with ARC or asthma in 63.8%; 44.8% of subjects without any atopic disease reported FH. Kiwi fruit caused symptoms most frequently (38.4%), followed by milk (32.6%), apple (29.1%), tomato (27.9%), citrus fruits (25.0%), tree nuts (23.3%), and peanut (17.4%). A total of 720 separate symptoms to 25 food items were reported. OAS was most common (51.2%), followed by gastrointestinal symptoms (23.5%), worsening of AD (11.4%), urticaria (4.2%), rhinitis or conjunctivitis (5.7%) and asthma (4.0%). Severe reactions occurred in 3.5% (25/720). Negative IgE and skin prick test to foods predicted well negative history, but the value of positive test results was limited. CONCLUSIONS: FH was reported most often by students with current AD and multiple atopic diseases. Severe reactions occurred especially in patients with ARC and asthma. After excluding lactose intolerance, milk hypersensitivity was frequently reported. PMID- 12752589 TI - CD63 expression on basophils as a tool for the diagnosis of pollen-associated food allergy: sensitivity and specificity. AB - BACKGROUND: Basophil activation is associated with the expression of CD63. Because allergens can induce basophil activation by cross-linking specific IgE, increased CD63 expression has been proposed as a novel in vitro test for immediate type allergy. OBJECTIVE: We compared the CD63-based basophil activation test (BAT) in the diagnosis of allergy to carrot, celery and hazelnut with skin prick tests (SPT) and measurement of allergen-specific IgE. METHODS: Twenty-nine patients with a history of an oral allergy syndrome induced by carrot, celery or hazelnut (n = 20 for each allergen) and 20 controls were studied. SPT were performed with standardized and native carrot, celery and hazelnut extracts. Allergen-specific IgE was determined by the CAP FEIA method and basophil activation was determined by flow cytometry upon double staining with anti IgE/anti-CD63 mAb. RESULTS: SPT with native carrot, celery and hazelnut showed sensitivities of 100%, 100% and 90%, and specificities of 80%, 80% and 90%. SPT with commercial extracts of the same allergens gave sensitivities of 85%, 80% and 85%, and specificities of 80%, 80% and 90%. Sensitivity of allergen-specific IgE and the BAT for carrot, celery and hazelnut was 80% vs. 85%, 70% vs. 85%, and 80% vs. 90%, with corresponding specificities of 80% vs. 85%, 80% vs. 80%, and 95% vs. 90%. The cut-off for a positive BAT was 10% CD63+ basophils. Moreover, there was a positive correlation between IgE reactivity and the number of CD63+ basophils for all food allergens (carrot: r = 0.69, celery: r = 0.67, hazelnut: r = 0.66). CONCLUSIONS: Quantification of basophil activation by CD63 expression is a valuable new in vitro method for diagnosis of immediate type food sensitization. Although double-blind placebo-controlled food challenges remain the gold standard, the CD63-based BAT may supplement routine diagnostic tests such as SPT or allergen-specific IgE in the future. PMID- 12752590 TI - Cytokine production by cord blood mononuclear cells stimulated with cows milk proteins in vitro: interleukin-4 and transforming growth factor beta-secreting cells detected in the CD45RO T cell population in children of atopic mothers. AB - BACKGROUND: Food antigens from the maternal circulation may sensitize fetal T cells in utero and be an important determinant in the development of food allergy. METHODS: Here we have examined the spontaneous and recall response to cow's milk proteins of cord blood mononuclear cells (CBMC) of newborn children, using single cell ELISPOT assays. RESULTS: In term newborns, confirming previous studies, the spontaneous cytokine response of CBMC is dominated by IL-4, IL-5, IL 10, and as shown here for the first time, TGF-beta. For TGF-beta only, the response of samples from infants of atopic mothers was significantly lower than samples from infants of non-atopic mothers. In vitro stimulation of CBMC with bovine serum albumin, casein and beta-lactoglobulin resulted in a significant increase of all cytokine-secreting cells, again dominated by T helper type 2 (Th2) cytokines. There was a clear tendency for samples from infants of atopic mothers to have lower Th2 responses than samples from infants of non-atopic mothers, which was particularly significant for both IL-4 and TGF-beta. Spontaneous cytokine secreting cells were virtually absent in cord blood from infants < 34 weeks gestation, as were cows milk protein-induced responses, although they were readily detectable in samples from infants aged > 34 weeks. To explore whether the cytokine secreting cells were in the naive CD4+ CD45RA population or memory CD4+ CD45RO T cells, these subsets were purified by positive and negative selection and tested for spontaneous and cows milk protein-induced cytokine responses. Strikingly, although the responses were small, the CD45RO+ cells from children of atopic mothers showed significant spontaneous and antigen specific IL-4 and TGF-beta responses, whereas the same population from infants of non-atopic mothers showed virtually no response. In addition CD45RA+ cells from infants of mothers with maternal atopy showed decreased IL-4 and TGF-beta responses, especially the latter. CONCLUSIONS: The cows milk antigen-specific IL 4 and TGF-beta responses preferentially seen in the memory cell subset of infants with a maternal history of atopy strongly suggests Th2 skewing to dietary antigens in utero. PMID- 12752591 TI - In vitro effects of eicosanoids derived from different 20-carbon fatty acids on T helper type 1 and T helper type 2 cytokine production in human whole-blood cultures. AB - BACKGROUND: Different series prostaglandins (PGs) and leukotrienes (LTs) are synthesized from different 20 carbon fatty acid precursors. The effects of the different series of PGs and LTs on production of T helper type 1 (Th1) and Th2 cytokines by human cells are not well established. OBJECTIVE: To characterize the influence of PGs and LTs produced from different fatty acid precursors on the Th1 and Th2 cytokine profile in mitogen-stimulated human whole-blood cultures. METHODS: Blood from healthy adult males was diluted and cultured with concanavalin A in the presence or absence of a range of concentrations of various PGs or LTs. Cytokine concentrations in culture supernatants were analysed by enzyme-linked immunosorbent assay (ELISA). RESULTS: PGE1, PGE2 and PGE3 significantly and dose-dependently decreased the concentrations of the Th1 cytokines IL-2 and IFN-gamma by up to 50% and 70%, respectively. The three PGs exhibited similar potency towards IFN-gamma production. At the highest concentration used (10-6 m) PGE1, but not PGE2 or PGE3, increased the concentration of the Th2 cytokine IL-4 by about 70%. IL-10 production was not affected by PGs. The ratio of the concentrations of IFN-gamma to IL-4 was significantly decreased at PGE concentrations of 10-7 and 10-6 M with all three PGEs having similar effects. LTB4, LTC4 and LTC5 did not significantly affect production of the cytokines studied. CONCLUSION: PGE produced from different fatty acids significantly decrease Th1 cytokine production resulting in a shift in the Th1, Th2 balance in favour of a Th2 response. PGE produced from different fatty acid precursors are equipotent in their effects on human T lymphocytes. Thus, although changes in the pattern of dietary fatty intakes may contribute to the increased prevalence of atopic disease, this would probably not be mediated through substitution of one PGE with another from a different series. It may, however, be mediated through a change in the total amount of PGE produced at the site of antigen presentation. PMID- 12752592 TI - Differences in circulating dendritic cell subtypes in cord blood and peripheral blood of healthy and allergic children. AB - BACKGROUND: Different types of circulating dendritic cells have been described. Dendritic cells influence differentiation of naive T lymphocytes into T helper type 1 (Th1) and Th2 effector cells. OBJECTIVE: The purpose of this study was to evaluate the number of circulating DC subtypes in peripheral blood of allergic and healthy children and in cord blood of neonates from allergic and non-allergic parents. METHODS: Circulating dendritic cells were flow cytometrically identified in whole blood samples as lineage (CD3, CD14, CD16, CD19, CD20, CD56) negative, CD34 negative and HLA-DR-positive cells. According to the expression of CD123 and CD11c, different DC subtypes were identified. RESULTS: Apart from DC1 (CD11c+ CD123dim+) and DC2 (CD11c- CD123high+), a third DC population was described with less differentiated phenotypic characteristics, namely CD11c- CD123dim+, and therefore defined here as less differentiated DC (ldDC). These ldDC represented the major DC population in cord blood and showed an age-depended decrease. The highest level of ldDC was detected in children with atopic dermatitis, whereas asthmatic children showed the lowest ldDC counts. Furthermore, high-dose inhaled corticosteroid treatment in asthmatic children was related to a decreased ldDC number. The number of circulating DC2 was significantly lower in allergic children, especially in asthmatics, compared to healthy children. In cord blood, no differences in DC subtypes were detectable between neonates at low and high risk for allergic disorders. CONCLUSION: These results indicate that, apart from DC1 and DC2, a third population of dendritic cells, identified as CD11c- CD123dim+ cells and defined as less differentiated DC, must be considered in the evaluation of circulating DC. Furthermore, DC2 counts were decreased in allergic children, especially in asthmatics, which might be the consequence of an increased recruitment to the target organs. PMID- 12752593 TI - Granule protein changes and membrane receptor phenotype in maturing human eosinophils cultured from CD34+ progenitors. AB - BACKGROUND: Eosinophils are now recognized as major effector cells in allergic and asthmatic disease with a potent armoury of mediators whose release makes a major contribution to the inflammation underlying these conditions. OBJECTIVES: The purpose of this study was to compare cultured eosinophils (CE) with normal density peripheral blood eosinophils (PBE) in terms of their membrane receptor expression and to analyse the expression and storage of the eosinophil granule proteins major basic protein (MBP) and eosinophil cationic protein (ECP) during eosinophil maturation in vitro. METHODS: Purified human peripheral blood CD34+ cells were cultured in the presence of recombinant human IL-3, IL-5, rhGM-CSF, SCF, and FLT-3 ligand. PBE were isolated by density gradient centrifugation and negative immunomagnetic selection. Expression of CD11b, CD18, CD45, CD45RA, CD45RB, CD45RO, CD69, CD95, IL-5Ralpha, IL-9Ralpha, CCR1, CCR3, and CXCR4 by CE as they matured in culture were assessed by immunostaining and flow cytometry and expression of these receptors compared with freshly isolated PBE. Immunohistochemical staining and labophot-2TM light microscopy determined expression of MBP, ECP, and CD69 during eosinophil maturation. RESULTS: Positive immunostaining for MBP and ECP was detectable in a proportion (15-20%) of CE as early as 3 days of culture even though these cells were mononuclear in appearance. The numbers of CE positive for both granule proteins increased in rhIL-3 and rhIL-5 treated cells to a maximum of approximately 80% by day 28. Maturing eosinophils exhibited positive immunostaining for CD69 after 14, 21 and 28 days of culture. Compared with PBE, CE had lower expression of pan-CD45 and CD45 isoforms, CD95 and CD11b. In contrast, the specific mean fluorescence for CD69, CD18, IL-5Ralpha, and IL-9Ralpha was significantly elevated for CE compared with PBE. CCR3 expression by CE and PBE was similar with no expression of CXCR4 detected by either CE or PBE. No significant difference in expression of CCR1 was found between CE and PBE. CONCLUSION: These data suggest that CE and PBE share many phenotypic properties and both MBP and ECP appear early in eosinophil development in vitro. However, there are quantitative differences that may be a consequence of their immaturity and/or the influence of the cytokines used in their culture. PMID- 12752594 TI - The production, storage and release of the neurotrophins nerve growth factor, brain-derived neurotrophic factor and neurotrophin-3 by human peripheral eosinophils in allergics and non-allergics. AB - BACKGROUND: Recent studies have shown that neurotrophins are produced by and can act on several immune-inflammatory cells. The origin of circulating as well as local neurotrophins is unknown. OBJECTIVES: The aim of this study was to assess whether eosinophils of allergic and non-allergic donors produce, store and release the neurotrophic factors NGF, BDNF and NT-3. METHODS: Eosinophils were purified by negative immunoselection (purity > 96%) from allergic asthmatics and non-allergic donors (25 to 53 years). The presence of mRNA for neurotrophic factors was evaluated by reverse transcription PCR. Specificity was demonstrated by cloning products and sequencing. Stored NGF, BDNF and NT-3 was demonstrated by Western-blotting and flow cytometry. Eosinophils were incubated and supernatants were collected for measurement of neurotrophic factors after cell stimulation with PAF. Neurotrophin content in eosinophil lysates was determined by ELISA. RESULTS: Eosinophils demonstrate mRNA for neurotrophins. Proteins were detectable by Western blot and FACS analysis. Neurotrophins were found in the eosinophil lysates at different amounts comparing allergic and non-allergic donors. Cell stimulation with PAF (10-8-10-5 M) after priming with GM-CSF leads to a dose dependant release of NGF and BDNF. CONCLUSIONS: Eosinophils store, produce and release NGF, BDNF and NT-3. They are a possible source of elevated neurotrophin levels found in allergy and asthma. PMID- 12752595 TI - Monomeric immunoglobulin E stabilizes FcepsilonRIalpha from the human basophil cell line KU812 by protecting it from natural turnover. AB - BACKGROUND: The high affinity IgE receptor (FcepsilonRI) on mast cells and basophils is up-regulated by its own ligand IgE; however, the mechanism is unknown. OBJECTIVE: To study the IgE-mediated effect on FcepsilonRI on basophils by using the human basophilic cell line KU812. METHODS: Expression of cell surface FcepsilonRI was assessed by flow cytometry. Western blot technique was used to illustrate tyrosine-phosphorylation and the Ca2+ level in KU812 was measured by fluorescence of Fura-2. Soluble specimens of the alpha-chain from FcepsilonRI (FcepsilonRIalpha) were obtained by lysing 107 KU812 pr. mL. FcepsilonRIalpha was detected by a sandwich immunoradiometric assay employing the IgE-binding capacity of FcepsilonRIalpha in conjunction with a monoclonal antibody. Polyclonal rabbit anti-FcepsilonRIalpha was used for detection of FcepsilonRIalpha by Western blotting. RESULTS: We found that monomeric IgE did not induce tyrosine-phosphorylation in KU812, which was the case when stimulating with IgE cross-linked by anti-IgE binding. Further, only cross-linking of IgE, but not monomeric IgE, increased the Ca2+ level. Using the immunoradiometric assay, we found a temperature dependent reduction in the amount of FcepsilonRIalpha. Samples incubated at 37 degrees C for 5 h displayed a 16-fold decrease in the FcepsilonRIalpha level compared with samples incubated at 4 degrees C. In the presence of IgE the reduction at 37 degrees C was only threefold. CONCLUSION: These results indicate that IgE does not induce intracellular signals in KU812, i.e., tyrosine-phosphorylation or Ca2+ release. Instead it appears that FcepsilonRIalpha is an unstable protein that IgE stabilizes and thereby protects from a temperature dependent turnover. PMID- 12752596 TI - Generation of monoclonal antibodies against Blo t 3 using DNA immunization with in vivo electroporation. AB - BACKGROUND: House dust mite allergy is closely associated with allergic diseases. Blomia tropicalis mite species is an important clinical species in the tropics. The cDNA clone encoding Blo t 3, a group 3 allergen from B. tropicalis, has been isolated in our laboratory. OBJECTIVE: This study was designed to generate Blo t 3-specific monoclonal antibodies (mAbs) for the detection, characterization and purification of this allergen. METHODS: Mice were immunized intramuscularly with naked plasmid DNA encoding Blo t 3 gene with in vivo electroporation. Hybridomas were generated by the fusion of the splenocytes to X63-Ag8.653 myeloma cells. Purified native Blo t 3 was obtained by mAb immuno-affinity purification and the allergenicity of native Blo t 3 was determined by human IgE enzyme-linked immunosorbent assay (ELISA). RESULTS: A panel of class-switched and high-affinity mAb recognizing a wide spectrum of Blo t 3 epitopes have been generated. These mAbs are useful for western immunoblot assay, sandwich ELISA and affinity purification of native Blo t 3. Allergenicity of native Blo t 3 protein was examined with 44 mite-allergic sera and approximately 57% of the tested sera had positive serum IgE reactivity to the native Blo t 3. CONCLUSIONS: These results demonstrated that intramuscular injection of naked DNA encoding Blo t 3 gene combined with in vivo electroporation is an effective and simple method to raise monoclonal antibodies that can be used for characterization and purification of Blo t 3. PMID- 12752597 TI - Characterization of immunoglobulin E responses in Balb/c mice against the major allergens of timothy grass (Phleum pratense) pollen. AB - BACKGROUND: Grass pollen, such as that from timothy grass (Phleum pratense), represents a major cause of type I allergy. OBJECTIVE: To characterize the IgE immune response and to identify the major allergens eliciting an IgE response in a mouse model using pollen extract of P. pratense for sensitization, in order to assess analogies to human hyperreactivity and to gain information on the allergenic potential as determined by the IgE-reactivity kinetics of defined allergens. METHODS: Balb/c mice were sensitized with pollen extract or with purified natural allergens. Serum IgE levels, the induction of specific IgE antibodies and immediate hypersensitivity were monitored by ELISA, Western blot and a skin test, respectively. RESULTS: The sensitized mice mounted a strong IgE response and showed IgE-reactivity first against Phl p 5a and 5b, then Phl p 4 and 13 and lastly against Phl p 6. No IgE response was mounted against Phl p 1. However, all purified fractions examined (Phl p 5a, 5b, 6 and 1) induced specific IgE and showed similar kinetics of IgE induction as pollen extract (first Phl p 5a and 5b, then Phl p 6). Skin test experiments demonstrated positive reactivity only in sensitized mice. CONCLUSION: The IgE reactivity induced by the major allergens in Balb/c mice was very similar to that found in allergic patients, with the exception of Phl p 1. The kinetics of the specific IgE response was comparable using either pollen extract or the purified major allergens, indicating that the intrinsic properties of the allergens are of importance rather than their proportionate amounts in pollen extract. This model should prove to be suitable for investigations regarding the mechanisms of induction and manifestation of timothy grass pollen allergy and for the evaluation of therapeutic strategies. PMID- 12752598 TI - Inhaled endotoxin and organic dust particulates have synergistic proinflammatory effects in equine heaves (organic dust-induced asthma). AB - BACKGROUND: Equine heaves is a naturally occurring organic dust-induced asthma characterized by airway neutrophilia, mucus hypersecretion and obstructive lung dysfunction. However, the relative role of different dust components in disease severity remains unclear. OBJECTIVE: This study investigated the relative contribution of inhaled endotoxin and organic dust particulates (mainly mould spores) in inducing heaves in heaves-susceptible horses. METHODS: Control and heaves-susceptible horses received inhalation challenges with hay dust suspension (HDS) before and after lipopolysaccharide (LPS) depletion. Heaves-susceptible horses also received inhalation challenge with HDS particulates with and without the addition of LPS and were housed in two separate dusty environments during which mould and endotoxin exposure was measured. The airway inflammatory and functional response to each challenge was measured. RESULTS: Depletion of endotoxin from HDS attenuated the airway neutrophilia and abrogated the airway dysfunction induced in heaves horses by inhaled HDS. The airway response was re established by adding back LPS to the depleted HDS, confirming that the attenuation in airway response was due specifically to endotoxin depletion. Interestingly, the magnitude of alteration in airway response following endotoxin depletion and add-back was greater than that which could be attributed solely to endotoxin per se, indicating that the LPS activity was enhanced by the other dust components. Consistent with this possibility, washed particulates harvested from HDS enhanced the airway response to inhaled LPS in heaves horses. Heaves horses given two different hay/straw challenges had a significantly different severity of airway inflammation and dysfunction, despite airborne dust and endotoxin concentrations in the horses' breathing zones being similar. CONCLUSION: Although inhaled endotoxin appears not to be the only determinant of disease severity in heaves, it does contribute significantly to the induction of airway inflammation and dysfunction. This contribution is largely via the synergistic action of inhaled endotoxin and organic dust particulates, although other soluble dust components also contribute to a lesser degree. PMID- 12752599 TI - Effect of lysed Enterococcus faecalis FK-23 (LFK) on allergen-induced peritoneal accumulation of eosinophils in mice. AB - BACKGROUND: The interest in anti-allergy immunoregulation by lactic acid bacteria has been growing for the last few decades. There is some evidence to suggest that lysed Enterococcus faecalis FK-23 (LFK) could relieve the clinical symptoms of pollinosis. However, the mechanism responsible for this phenomenon remains unknown. OBJECTIVE: To identify the effect of LFK, a lysozyme treated and heat killed preparation from the lactic acid bacteria Enterococcus faecalis FK-23 strain, on allergen-induced eosinophil accumulation. METHODS: BALB/c mice were sensitized with ragweed pollen extract, and peritoneal accumulation of eosinophils was induced. A total of 60 mg (0.5 mL) LFK was orally administered to the experimental mouse every day during 21 days of the sensitization period. In addition, LFK 4 mg, 25 mg and 60 mg (each 0.5 mL) were also orally administered to a mouse of each group every day for 21 days. Saline was fed in a dose of 0.5 mL/mouse per day for the same duration as a control. RESULTS: Compared with control mice, LFK-treated mice exhibited decreased ragweed pollen allergen induced peritoneal accumulation of eosinophils (P = 0.013), which showed a tendency to be in a dose-dependent fashion (P = 0.14). CONCLUSION: The results provide laboratory evidence of the role for LFK, a lactic acid bacteria preparation, in combating eosinophil accumulation. PMID- 12752600 TI - Dissociation of T helper type 2 cytokine-dependent airway lesions from signal transducer and activator of transcription 6 signalling in experimental chronic asthma. AB - BACKGROUND: Type 2 T helper lymphocytes (Th2 cells) and their cytokine products are important in the pathogenesis of asthma. OBJECTIVE: To examine the contribution of the signal transducer and activator of transcription (STAT) 6 pathway, involved in Th2 cytokine signalling, to the development of lesions of chronic asthma. METHODS: BALB/c mice sensitized to ovalbumin were chronically challenged by inhalational of low mass concentrations of antigen for 6 weeks. Airway lesions in wild-type mice were compared with those in STAT6-deficient mice and in IL-4/13 double-deficient mice by histomorphometry and immunohistochemistry. Airway responses to methacholine were evaluated by whole body plethysmography. Cytokine production by peribronchial lymph node cells was quantified by enzyme immunoassay. RESULTS: STAT6-/- mice developed a variety of airway lesions that were at least equivalent to those in wild-type mice, including accumulation of intraepithelial eosinophils and of chronic inflammatory cells in the lamina propria, subepithelial fibrosis and epithelial thickening. In addition, STAT6-/- mice exhibited exaggerated airway hyper-reactivity (AHR) compared to wild-type animals. This was despite a shift from a Th2 to a Th1 pattern of immunoglobulin production by plasma cells in the inflammatory infiltrate and diminished mucous cell hyperplasia/metaplasia, together with increased production of IFN-gamma by peribronchial lymph node cells, consistent with absence of signalling via the STAT6 pathway. In contrast, gene-targeted IL 4/13-/- mice exhibited markedly diminished eosinophil recruitment and airway remodelling, as well as absence of AHR. CONCLUSIONS: In this model, the effects of STAT6 deficiency were in marked contrast to the suppression of inflammation and AHR described in models of allergic bronchopulmonary inflammation. These results, which provide evidence of STAT6-independent AHR in an inhalational challenge model of chronic asthma, emphasize the critical effector roles of IL-4 and IL-13, as well as the need to use appropriate models to understand cytokine signalling pathways that may be potential therapeutic targets in asthma. PMID- 12752601 TI - Enhancement of gelatinase activity during development of subepithelial fibrosis in a murine model of asthma. AB - BACKGROUND: Chronic asthma is characterized by inflammatory cell infiltration and tissue remodelling leading to subepithelial fibrosis. Metalloproteinases (MMPs) are involved in degradation of extracellular matrix in most chronic inflammatory diseases. OBJECTIVE: The aim of this study was to investigate the expression of MMPs in the development of inflammatory processes associated or not with the concomitant development of subepithelial fibrosis in an experimental model of asthma. METHODS: Sensitized BP2 mice were challenged with ovalbumin (OA) every 2 weeks during 8 months. Several mice were removed once a month and bronchoalveolar lavages (BAL) or lung biopsies were performed. RESULTS: Lung sections stained with picrosirius and hydroxyproline measurements showed a significant collagen deposition after 16 weeks of OA challenge, demonstrating the development of subepithelial fibrosis. Pulmonary inflammation was present from the first OA challenge and was consistent throughout the 8 months of the study. Moreover, an up-regulation and activation of MMP-9 and, to a less extent, MMP-2 were observed in BAL fluid from challenged mice. The level of tissue inhibitor of metalloproteinases (TIMP)-1 increased after 12 weeks of OA challenge vs. control mice. CONCLUSION: This study reveals that a decrease in the activation of the MMP 9 due to the increase in TIMP-1, could contribute to excessive collagen deposition following repeated antigen challenge in sensitized mice. PMID- 12752602 TI - Mast cells play a partial role in allergen-induced subepithelial fibrosis in a murine model of allergic asthma. AB - BACKGROUND: Role of mast cells in the development of allergen-induced airway remodelling has not been fully investigated in vivo. OBJECTIVE: To clarify the possible role of mast cells in the development of allergen-induced airway remodelling, we compared their responses of genetically mast cell-deficient mice, WBB6F1-W/Wv (c-kit mutant) and Sl/Sld (c-kit ligand mutant) mice with those of congenic normal mice in a murine model of allergic asthma. METHODS: Mice were sensitized to ovalbumin (OA) with alum, and exposed daily for 3 weeks to aerosolized OA. Twenty-four hours after the last inhalation, bronchial responsiveness to acetylcholine (Ach) was measured, and bronchoalveolar lavage (BAL), and biochemical and histological examinations were performed. RESULTS: In both sensitized mast cell-deficient mice, the degree of bronchial hyper responsiveness to Ach, the number of inflammatory cells and the level of transforming growth factor-beta1 in BAL fluid, IgE response and goblet cell hyperplasia in the epithelium after repeated allergen provocation were not significantly different from those of congenic mice. In contrast, subepithelial fibrosis, evaluated in the fibrotic area around the airways, observed in congenic mice after repeated allergen challenge was partially attenuated in both mast cell deficient mice. In addition, the amount of hydroxyproline in the lung of mast cell-deficient mice was significantly lower than that of congenic mice. Furthermore, the decreased fibrotic area and amount of hydroxyproline in W/Wv mice was completely recovered by reconstitution of tissue mast cells with bone marrow-derived mast cells of congenic mice. CONCLUSION: These findings suggest that mast cells play a partial role in the development of allergen-induced subepithelial fibrosis, although airway inflammation, epithelial remodelling and BHR caused by repeated allergen challenge are independent of mast cells, at least in this model. PMID- 12752603 TI - Penicillin allergy. PMID- 12752606 TI - Who manages the care of students with Attention Deficit Hyperactivity Disorder (ADHD) in higher education? AB - OBJECTIVE: To identify who provides medical support to students with Attention Deficit Hyperactivity Disorder (ADHD) while in higher education. METHODS: A questionnaire pack was sent to 50 undergraduate student health centres attached to higher education colleges/universities. RESULTS: Eighty-two per cent of practices returned questionnaires. Forty-nine per cent had undergraduate students with ADHD, of those 76% were on methylphenidate. Fifty-two per cent saw only their general practitioner (GP) for follow up and the rest were jointly managed by GP with: psychiatrists, paediatricians, psychologists or physicians. Eighty seven per cent of GPs had not attended recent courses or training on ADHD. CONCLUSION: Those caring for undergraduate students with ADHD are still largely unfamiliar with the condition. Guidelines need to be drawn up to establish handover from paediatric to adult care. PMID- 12752607 TI - The effects of work on the growth of Jordanian boys. AB - OBJECTIVE: To examine the effects of work on growth among boys aged 10-16 years in Jordan. STUDY DESIGN: Cross-sectional health survey comparing working and non working boys. MAIN OUTCOMES: Height for age z-score; weight for age z-score. METHODS: One hundred and thirty-five working and 405 non-working boys aged 10-16 years were studied in the Jordanian areas of Irbid, Jarash and North Jordan Valley. The boys and their mothers were interviewed and data collected on work status, child's smoking status, and family socio-economic characteristics including per capita income, family size, maternal and paternal education, area of the house in square metres and expenditure on household durables. Height and weight were obtained at the time of interview. Linear regression models were fitted on height for age z-score and weight for age z-score. RESULTS: In bivariate analysis, child's work status (r = -0.221), household per capita income (r = 0.104), family size (r = -0.102), house m2 per capita (r = 0.090) and monthly expenditure on durables (r = 0.086) were significantly correlated with height z-score. Work status (P < 0.0001) and household m2 per capita (P = 0.002) were retained in the regression model fitted on height z-score which explained 9.8% of the variance. The model fitted on weight z-score explained 6.5% of the variance and work status (P < 0.0001), household per capita income (P = 0.041) were retained. CONCLUSIONS: The results of this study suggest that, independent of a range of socio-economic factors, work has a negative effect on child growth. Given the extent of child labour internationally these findings have profound implications for global child well-being and for global social and economic policy. PMID- 12752608 TI - Child health profiles: the viewpoints of adults. AB - THE CHILD HEALTH PROFILE: The teenage years have been identified as an important time to influence attitudes and behaviour likely to affect current and future health. The child health profile was developed partly to provide appropriate health information to young people aged 10 and older, but was also intended to recognize their right to assume greater responsibility for their health records. It was seen as an extension of the popular parent held record for younger children. METHODS: A Scottish child health profile was introduced through schools in three health board areas. Data about usage and views were obtained from children and relevant adults by means of questionnaires. In this paper, the perspectives of parents, health professionals and teachers are reported. FINDINGS: The majority of the adults were keen on the idea of the profile in principle. However, most parents knew little about it, even though all their children had been issued with one. Teachers had little involvement with the profile and the majority of health professionals had some doubts about its application. The adults indicated that a small number of children made effective use of the profile, but information from both adults and children indicated that it was seldom discussed during consultations with health professionals, few of whom routinely asked to see it. CONCLUSIONS: The paper outlines implications for the provision of health information and records for this age group. PMID- 12752609 TI - Infant-mother interaction as a predictor of child's chronic health problems. AB - BACKGROUND: Psychological stress is associated with physical illnesses like asthma or infections. For an infant, situations perceived as stressful are highly dependent on the relationship with the caregiver. Constantly poor mother-infant interaction increases the child's vulnerability to stressful conditions and experiences. The aim of the study was to investigate the impact of the quality of early mother-infant interaction on the subsequent physical health of the child. Poor mother-infant interaction was hypothesized to be associated with chronic or recurrent health problems in the child. PARTICIPANTS: Fifty-seven mother-infant dyads from families at risk of psychosocial problems and 63 from non-risk families, altogether 120 dyads, participated in the study. Families were drawn from normal population, from well-baby clinics in the city of Tampere, Finland. Infants were full-term and healthy, families with severe risks like psychotic illnesses of the parents or a history of child protection concerns were excluded from the study. METHODS: After the initial interview with the mother, the mother infant interaction was videotaped when the infants were 8-11 weeks of age and the interaction was assessed using the Global Rating Scale for Mother-Infant Interaction (Murray et al. 1996a). After the 2-year follow-up mothers were interviewed again and the health problems of the child were elicited. RESULTS: Poor dyadic mother-infant interaction and infant's poor interactive behaviour assessed at two months were separately associated with the physical health of the child during the two-year follow-up. After adjusting for other factors in the logistic regression analysis infant's poor interactive behaviour remained as a significant predictor of chronic or recurrent health problems in the child. Infant's health problems at the time of the initial interview and day care centre attendance were also significant predictors. CONCLUSIONS: The results suggest that interactional issues between a mother and her infant are related to the child's subsequent physical health. Children with recurrent or chronic health problems may have relationship difficulties with which they need help. Also, early avoidant behaviour of the infant should be regarded as an indicator of the infant's distress with possibly adverse outcomes in the child's physical health, among other consequences. PMID- 12752610 TI - Behavioural problems, social competence and self-concept in siblings of children with autism. AB - INTRODUCTION: This study investigated the psychological adjustment of siblings of children with high-functioning autism (HFA) in comparison with siblings of normally developing children in the domain of behavioural problems, social competence and self-concept. METHOD: Twenty-nine siblings of children with HFA and 29 siblings of children without a disorder participated in the study. Standardized, written questionnaires were used. RESULTS: Siblings of children with HFA, especially brothers and sisters between 6 and 11 years old, had more behavioural problems than siblings of the control group. Sisters of children with HFA ascribed higher social competence to themselves. Sisters of children with HFA between 12 and 16 years old had a more positive self-concept. In both groups siblings with a more negative self-concept had less social skills and siblings with a more positive self-concept scored better in the social domain. Finally, in accordance with the control group, the perception of the siblings' social competence of parents of children with HFA broadly matched the perception siblings had of themselves. DISCUSSION: Overall, siblings of children with HFA are not more susceptible to adaptation problems than siblings of children without a disorder. Implications for practice and further research are discussed. PMID- 12752611 TI - When adolescents disagree with their mothers: CBCL-YSR discrepancies related to maternal depression and adolescent self-esteem. AB - OBJECTIVE: To facilitate understanding of disagreement between mothers and adolescents when they report on adolescents' mental symptoms, discrepancy of reports were studied in relation to depressive symptoms of mothers and self esteem of adolescents. PARTICIPANTS: Sixty-eight mother-adolescent pairs participated. All the adolescents (11-17 years) were referred to child and adolescent psychiatric outpatient clinics in Norway for emotional or behavioural disorders. METHODS: The adolescents completed the questionnaire Youth Self Report (YSR), and mothers completed the corresponding Child Behaviour Checklist (CBCL). Maternal depression was measured with the Hospital Anxiety and Depression Scales and adolescent self-esteem was measured with Self-Perception Profile for Adolescents. RESULTS: Mothers' level of depression emerged as a significant variable (P < 0.001) predicting CBCL-YSR discrepancy on internalizing disorders, explaining 41% of the variance. With increased levels of depression mothers tended to report more internalizing problems compared to the adolescents. For mother-adolescent discrepancy on externalizing problems, adolescents' age was the most significant variable. With increasing age the adolescents were more in agreement with their mothers when reporting their externalizing problems. But also self-esteem problems concerning their looks made adolescents inclined to admit more externalizing problems, thus being more in agreement with their mothers. CONCLUSIONS: Subjective, psychological variables such as mothers' level of depression and self-esteem of adolescents may be useful to consider when interpreting informant discrepancy concerning the mental problems of adolescents. PMID- 12752612 TI - Current practice in the management of Attention Deficit Disorder with Hyperactivity (ADHD). AB - BACKGROUND: The management of Attention Deficit Disorder with Hyperactivity (ADHD) is controversial and guidance published by the National Institute for Clinical Excellence (NICE) suggests a multidisciplinary approach. METHODS: We evaluated the patterns of referral and techniques used in the diagnosis and management of ADHD in Wirral Health Authority by studying the case notes of 391 children diagnosed between 1994 and August 2000. RESULTS: The majority of cases were referred from Primary Care and the detected prevalence was 8/1000. Boys were diagnosed more frequently than girls (12 : 1) and the mean age at diagnosis was 8.7 years; girls were more likely to have been diagnosed prior to age 8. The use of behaviour rating questionnaires and school visits as an aid to diagnosis increased during the study period. Management by Child Psychiatry and Community Paediatric services was similar with stimulant medication prescribed in 91% of cases; Child Psychiatry services were more likely to give advice on behaviour management than Paediatric services. Twelve per cent of cases were managed by both services, or in joint Outpatient clinics. Outpatient follow-up was most likely to occur at six monthly intervals and required 1312 appointments per year. CONCLUSIONS: This study shows that the management of children with ADHD in Wirral Health Authority is compatible with NICE guidance. Attention Deficit Disorder with Hyperactivity is a significant burden on clinical services and long-term planning is required. PMID- 12752613 TI - Informing parents of visually impaired children: who should do it and when? AB - BACKGROUND: Parents of sick or disabled children are likely to be more stressed than parents of non-disabled children and may benefit from being given information about their child's condition and its implications, but the stage at which parents should receive such information and who should provide it has not been fully investigated. The impact of written information on stress levels of parents of visually impaired children, seen in the ophthalmology clinic of a children's hospital, was explored in this study and the question of who parents thought should provide information, and at what stage, was also investigated. METHODS: A longitudinal, experimental intervention study was conducted to compare the effect on perceived stress levels of providing information about the implications of visual impairment to parents. Effects were compared according to the child's age and explored in relation to baseline stress levels. The study included a frequency analysis about parents' knowledge of visual impairment and how it relates to education, and their response to being given written information. RESULTS: The results did not show an effect on levels of parental stress but did find that parents of school age children were more stressed than those of preschool age. Over 80% of participants considered that information was given too late and suggested it should be given soon after diagnosis. Of the controls, 32.6% thought the general practitioner should provide information on education although participants were more likely to expect the hospital to provide it. CONCLUSIONS: The majority of parents would prefer to receive information soon after diagnosis of their child's visual impairment. Most parents do not know who to approach for information on education. Giving them access to patient liaison teams who could advise about the repercussions of visual impairment in children would be beneficial. PMID- 12752620 TI - Core needle biopsy versus fine-needle aspiration biopsy: are there similar sampling and diagnostic issues? PMID- 12752621 TI - Review of 125 SiteSelect stereotactic large-core breast biopsy procedures. AB - Advances in stereotactic breast biopsies have introduced a variety of devices that yield different sizes of tissue samples. The choice of biopsy device should be based on which technique is most likely to yield a definitive diagnosis at the time of the initial biopsy. This is a prospective study of 104 patients who underwent a total of 125 stereotactic breast biopsies using the SiteSelect large core biopsy device. From May 1999 to June 2001, 104 patients underwent 125 stereotactic breast biopsies with the SiteSelect large-core biopsy device. One hundred four 15 mm SiteSelect biopsies, eighteen 10 mm SiteSelect biopsies, and three 22 mm SiteSelect biopsies were performed. Atypical ductal hyperplasia (ADH) and ductal carcinoma in situ (DCIS) were found in 15% of the biopsies and infiltrating cancer was found in another 15% of the biopsies. Seventy-eight percent of the ADH and 90% of the DCIS lesions were associated with indeterminate calcifications noted on mammogram. Two of the 22 mm SiteSelect excisions yielded a specimen that contained the entire cancer with clear surgical margins. All of the patients with DCIS or invasive carcinoma underwent definitive surgical and adjuvant therapy. The sensitivity and specificity of SiteSelect in this series of patients was 100%. The SiteSelect biopsy procedure is safe, well tolerated by patients, and can be performed under local anesthesia. SiteSelect is comparable to an open excisional biopsy in its ability to obtain adequate tissue for accurate diagnosis, but excises significantly less normal surrounding breast tissue. Based on the data, indications for primary use of SiteSelect are indeterminate calcifications on mammogram, rebiopsy of a vacuum-assisted biopsy site that yielded atypia on pathologic examination, and complete excision of a lesion suspicious for invasive carcinoma in order to assess actual size and margin status. PMID- 12752622 TI - Clinicopathologic analysis of sentinel lymph node mapping in early breast cancer. AB - Axillary nodal status is the most significant prognosticator for predicting survival and guiding adjuvant therapy in breast cancer patients. Sentinel lymph node biopsy (SLNB) represents a minimally invasive procedure with low morbidity for staging axillary nodal status. In this article we review and report our experiences in patients with early breast cancer who underwent SLNB at the Revlon/UCLA Breast Center. Between September 1998 and May 2000, a total 83 SLNBs were performed in 81 patients with proven breast cancer and negative axillary examination who elected to have SLNB as the first step of nodal staging. Two patients had bilateral breast cancer. SLNB was localized by using both 99Tc sulfur colloid (83 cases) and isosulfan blue dye (75 cases). Data of these patients were prospectively collected and analyzed. The clinical and pathologic characteristics of women with positive and negative sentinel lymph nodes (SLNs) were compared to identify features predictive of SLN metastasis. Of the 83 cases, the SLN was successfully localized in 82 (98.8%). Sixty-three percent of patients had SLNs found in level I only, 18.3% in both level I and II, and 4.9% in level II alone. The vast majority (84.3%) of these cases had T1 breast cancer with an average size of 1.55 cm for the entire series. Twenty-three patients (28%) had positive SLNs, with an average of 1.5 positive SLNs per patient. Fifteen had metastases detected by hematoxylin and eosin staining and 8 had micrometastases detected by immunohistochemistry (IHC) using anticytokeratin antibodies. Ten of the former group agreed to and 2 of the latter group opted for full axillary lymph node dissection (ALND). An average of 17.5 lymph nodes were removed from each ALND procedure. Additional metastases or micrometastases were found in seven patients (in a total of 28 lymph nodes). Three patients with completely negative SLNs experienced additional axillary lymph node removal due to their election of free flap reconstruction. None had metastases detected in these lymph nodes. The absence of estrogen and progesterone receptors (ER/PR) by IHC (p = 0.036) and the presence of lymphatic/vascular invasion (LVI) (p = 0.002) predicted positive SLNs in patients with early breast cancer in a univariate analysis; in a multivariate analysis only LVI was predictive (p = 0.0125). Histologic type, nuclear grade, tumor differentiation, HER-2/neu and p53 status, S-phase fraction, and DNA ploidy did not predict SLN status. Immediate postoperative complications were uncommon and delayed complications completely absent. Because of the high detection rate, accurate staging, and minimal morbidity, SLNB should be offered as a choice to women with small breast cancers and clinically negative nodes. Because positive LVI and negative ER/PR status are highly predictive of pathologically positive SLNs in small breast cancers, women whose cancers meet these criteria should be advised preoperatively about their risk of having a positive SLN and may benefit from intraoperative assessment (frozen section and/or touch preparation) of their SLNs. PMID- 12752623 TI - Method for combined FDG-PET and radiographic imaging of primary breast cancers. AB - The purpose of the study was to demonstrate the feasibility of a hybrid functional/anatomic breast imaging platform with biopsy capability for facilitating lesion detection and diagnosis. This platform consists of an investigative dedicated positron emission mammography (PEM) device mounted on a stereotactic X-ray mammography system, permitting sequential acquisition of mammographic and emission images during a single breast compression. There is automatic coregistration of images from both modalities, and these results can be successfully correlated with histopathologic findings. The potential utility of functional images correlated to anatomic images would include noninvasively detecting clinically and radiographically occult cancers, assessing response to therapy, discriminating between benign and malignant breast masses, and ultimately reducing the number of invasive and costly surgical interventions. A spot-digital mammogram and subsequent PEM image, collected over a 4-minute period, were obtained in a single patient with the breast in compression after intravenous injection of (F-18)-2-deoxy-2-fluoro-D-glucose (FDG) at the time of stereotactic biopsy. The authors conclude that FDG-based lesion localization information may be combined with the lesion X-ray attenuation characteristics using this common imaging platform. PMID- 12752624 TI - BRCA status, molecular markers, and clinical variables in early, conservatively managed breast cancer. AB - Conservatively treated premenopausal breast cancer has a higher rate of local relapse as well as an increased genetic predisposition to cancer. The current study's purpose was to evaluate the interactions between BRCA-1/2 status and molecular biologic markers in a cohort of conservatively managed breast cancer patients. Seventy-six premenopausal women treated with breast-conserving surgery and radiation therapy were this study's focus. All patients were treated with wide local excision with or without axillary dissection, followed by radiation to the intact breast. Systemic therapy was administered as clinically indicated. All patients in this study had an available paraffin block from the primary tumor and agreed to undergo complete sequencing of the BRCA-1 and BRCA-2 genes. The primary breast tumor tissue from each patient was immunohistochemically stained for estrogen receptor (ER), progesterone receptor (PR), p53, HER-2/neu, and Proliferating Cell Nuclear Antigen (PCNA). Of the 76 patients tested for BRCA, 50 patients had wild-type BRCA-1 and BRCA-2, 15 had variants of unclear significance, 6 had deleterious mutations in BRCA-1, and 5 had deleterious mutations in BRCA-2. p53 positivity correlated with deleterious mutations in BRCA 1 (p = 0.023), but not in BRCA-2. Though not significant, there was a trend for ER and PR negativity to correlate with BRCA-1 mutation (p = 0.087 and 0.054, respectively); there were no correlations between ER, PR, and BRCA-2. Though not significant, all 11 tumors with BRCA mutations were HER-2/neu negative. Patients with BRCA mutations have a unique molecular profile. These data can be helpful in understanding differences in the biologic behavior of patients with familial breast cancers. PMID- 12752625 TI - Effects of screening mammography on the comparative survival rates of African American, white, and Hispanic beneficiaries of a comprehensive health care system. AB - This study reviewed mammographic screening related to breast carcinoma diagnosis and treatment between 1987 and 1997 at Brooke Army Medical Center, San Antonio, TX. Epidemiologic data from the Department of Defense Automated Central Tumor Registry were merged with data from patients' medical records and responses of the patients or their families to a mailed survey. The cases of 907 women grouped by race-white, African American, and Hispanic-were analyzed. Breast carcinoma diagnosed by mammographic screening showed a reversed ratio of early to late stage of cancer occurring for all three groups. That ratio was 1.45 for African Americans, 2.67 for Hispanics, and 3.08 for whites. For those women diagnosed with screening mammography, no statistically significant difference in 5-year survival was found between the races: 86% for whites, 83% for Hispanics, and 80% for African Americans. Mammographic screening as a diagnostic tool appears to equalize survival among whites, Hispanics, and African Americans, in spite of differences in age, stage of diagnosis, and military rank used as a proxy for socioeconomic status. When not controlling for mammographic diagnosis, Kaplan Meier analysis revealed significant differences in survival patterns between whites, Hispanics, and African Americans. Five-year survival rates were 71% for whites, 74% for Hispanics, and 53% for African Americans. Screening mammography reduced 5-year mortality by almost 59% in African Americans, 52% in whites, and 36% in Hispanics. Whites were diagnosed with breast carcinoma, on average, at 57 years of age-11 years later than African Americans (average age 46 years) and 7 years later than Hispanic women (average age 50 years). As a diagnostic tool, screening mammography was used to discover breast cancer in 36% of white women, 33% of Hispanics, and 22% of African Americans. Further research is recommended to examine the use of mammography among various racial/ethnic groups. PMID- 12752626 TI - Volatile markers of breast cancer in the breath. AB - Breast cancer is accompanied by increased oxidative stress and induction of polymorphic cytochrome P-450 mixed oxidase enzymes (CYP). Both processes affect the abundance of volatile organic compounds (VOCs) in the breath because oxidative stress causes lipid peroxidation of polyunsaturated fatty acids in membranes, producing alkanes and methylalkanes which are catabolized by CYP. We performed a pilot study of breath VOCs, a potential new marker of disease in women with breast cancer. This was a combined case-control and cross-sectional study of women with abnormal mammograms scheduled for a breast biopsy. Breath samples were analyzed by gas chromatography and mass spectroscopy in order to determine the breath methylated alkane contour (BMAC), a three-dimensional display of the alveolar gradients (abundance in breath minus abundance in room air) of C4-C20 alkanes and monomethylated alkanes. BMACs in women with and without breast cancer were compared using forward stepwise discriminant analysis. Two hundred one breath samples were obtained from women with abnormal mammograms and biopsies read by two pathologists. There were 51 cases of breast cancer in 198 concordant biopsies. The breath test distinguished between women with breast cancer and healthy volunteers with a sensitivity of 94.1% (48/51) and a specificity of 73.8% (31/42) (cross-validated sensitivity 88.2% (45/51), specificity 73.8% (31/42)). Compared to women with abnormal mammograms and no cancer on biopsy, the breath test identified breast cancer with a sensitivity of 62.7% (32/51) and a specificity of 84.0% (42/50) (cross-validated sensitivity of 60.8% (31/51), specificity of 82.0% (41/50)). The negative predictive value (NPV) of a screening breath test for breast cancer was superior to a screening mammogram (99.93% versus 99.89%); the positive predictive value (PPV) of a screening mammogram was superior to a screening breath test (4.63% versus 1.29%). A breath test for markers of oxidative stress accurately identified women with breast cancer, with an NPV superior to a screening mammogram. This breath test could potentially be employed as a primary screen for breast cancer. Confirmatory studies in larger groups are required. PMID- 12752627 TI - Local and regional therapy for women with breast cancer in British Columbia. AB - British Columbian provincial practice guidelines (PPGs) have recommended breast conserving surgery (BCS), axillary node dissection, and radiation therapy following BCS for specific subgroups of breast cancer patients. Patient-, disease , and physician-specific factors associated with these therapies were investigated in nonmetastatic invasive breast cancer patients. Temporal trends in BCS and physicians' experiences with PPGs were also examined. Sources of data for patient, disease, treatment, and treating physician factors included medical records, source documents, and the British Columbia Medical Directory for 967 nonmetastatic invasive breast cancer patients diagnosed in British Columbia in 1995. BCS utilization among 496 patients with pathologically node-negative breast cancer (NNBC) was compared to earlier British Columbian data. Family physicians and surgeons were surveyed in 1997 regarding their experience with PPGs. 57% of "ideal" candidates received BCS; 87% of patients received axillary node dissection; and 95% of women treated with BCS also received radiation therapy. Tumor size, tumor location, and extent of ductal carcinoma in situ (DCIS) were associated with BCS use; age, tumor size, and tumor location were associated with axillary node dissection; and age alone was associated with radiation therapy following BCS. Fifty-four percent of NNBC patients received BCS in 1995, compared to 44% in 1991, with increases seen in most patient-, disease-, and physician specific comparisons. The increase in BCS, and high proportion completing radiation therapy, are encouraging and may be due in part to greater exposure to PPGs. PMID- 12752628 TI - Microvessel density and vascular endothelial growth factor expression in infiltrating lobular mammary carcinoma. AB - Angiogenesis is an important prognostic factor in infiltrating ductal carcinoma (IDC). Vascular endothelial growth factor (VEGF) stimulates angiogenesis in vivo. VEGF expression has been correlated with high vascularity in IDC. However, little is known about the prognostic significance of microvessel density (MVD) and its correlation with the expression of VEGF in infiltrating lobular carcinoma (ILC). We analyzed tumor samples from 51 patients with primary classic ILC to determine the relationship between tumoral MVD and VEGF expression. Cases of pleomorphic lobular carcinoma and tubulolobular carcinoma were excluded. Five-micron thick sections obtained from formalin-fixed, paraffin-embedded tissue blocks were immunostained with antibodies to factor VIII-related antigen (Dako, Carpenteria, CA) and VEGF (Calbiochem, Boston, MA). The former was used for MVD analysis. The vessel counts from the three most vascular fields (x200 magnification) were recorded and the highest of the vessel counts of the three fields was designated as the MVD. The intensity of VEGF staining and the proportion of cells staining were scored. Both the vessel counts and the scoring of VEGF staining were evaluated by two independent pathologists. The Student's t-test and Wilcoxon rank sum test were used to compare mean MVD and VEGF scores according to various clinical and pathologic features. All significance tests were two-sided with an alpha-level of 0.05. There was good correlation between the MVD of each observer (correlation coefficient 0.775, p < 0.001). There was no correlation of MVD or VEGF score with the size or stage of the tumor. In addition, the MVD or VEGF score was not significantly different between axillary lymph node-positive cases and node-negative cases, between patients with recurrence and those without, and between patients who survived and those who died of disease. There was, however, a weak negative correlation between the MVD and VEGF expression (Spearman correlation coefficient -0.08). Neither MVD or VEGF immunoscore were associated with tumor recurrence or vital status in patients with ILC. The absence of a statistically significant positive correlation between MVD and VEGF expression suggests that other factors may play a more important role in the angiogenesis of ILC. PMID- 12752629 TI - Male patients with diagnoses of both breast cancer and prostate cancer. AB - Recently cancer susceptibility syndromes have been characterized that suggest possible genetic linkages between breast cancer and prostate cancer within families. Despite these connections, male breast cancer and prostate cancer in an individual man has rarely been reported. The clinical features of 10 patients with both of these cancers are described here. One hundred sixty-one patients with male breast cancer were seen at the Dana-Farber Cancer Institute and Massachusetts General Hospital between 1977 and 2000. Of these, 10 were identified who also had prostate cancer. A retrospective review of records from these 10 patients was performed. Breast cancer preceded prostate cancer in eight of these men. The mean age of diagnosis of breast cancer was 65.7 years (range 47 72 years). Twenty percent had nodal involvement at diagnosis and two patients ultimately developed evidence of metastatic disease. The mean age of diagnosis of prostate cancer was 68.0 years (range 51-76 years) with a median prostate specific antigen (PSA) level at diagnosis of 6 ng/ml (range 1.8-47.5 ng/ml). Seven patients had a family history of female breast cancer in a first-degree relative, while one had a family history of prostate cancer. At a median follow up of 6.5 years from initial cancer diagnosis, one patient had died of metastatic breast cancer and another had died of metastatic prostate cancer. The clinical features and course of the breast cancers diagnosed in this series do not appear significantly different from those described for the general population of male breast cancer patients. In addition, these men do not appear to develop prostate cancer at an earlier age or more aggressive stage than the general population. PMID- 12752630 TI - A roundtable discussion of aromatase inhibitors as therapy for breast cancer. AB - This article summarizes the conclusions of a meeting of diverse breast cancer experts who discussed issues, controversies, and new clinical trial results relevant to the use of aromatase inhibitors for treating postmenopausal women with breast cancer. The new generation of aromatase inhibitors (anastrozole, letrozole, exemestane) have largely replaced megestrol acetate as a second-line therapy in postmenopausal women with hormone-responsive advanced breast cancer. In addition, anastrozole and letrozole have been shown to be superior to tamoxifen for first-line therapy. Finally, recent results suggest that anastrozole may be superior to tamoxifen as adjuvant therapy for early stage disease in postmenopausal women with hormone-responsive disease. PMID- 12752631 TI - Nodular fasciitis of the breast: a case report. AB - Nodular fasciitis is a soft tissue lesion that in rare instances occurs in the breast. It can clinically and radiologically mimic malignant tumor. We describe a case of nodular fasciitis of the breast in a young woman that was misdiagnosed as phyllodes tumor. The histologic features and a review of the literature are presented. Awareness of such an entity in the breast obviates the need for unnecessary surgical intervention. PMID- 12752632 TI - Clip placement after sonographically guided percutaneous breast biopsy. AB - A simple and inexpensive technique for deployment of a metallic marker at the site of an ultrasound guided core breast biopsy is described. An illustrative case in which this technique was employed to mark the location of three biopsied lesions is presented. PMID- 12752633 TI - Primary osteosarcoma of the breast: a case report. AB - We report a rare case of primary osteosarcoma of the breast in a postmenopausal patient without any association to either trauma or an underlying tumor. Clinical, radiographic, and histologic illustrations as well as a review of the literature are presented. PMID- 12752634 TI - An axillary recurrence of breast cancer following a negative sentinel lymph node biopsy. AB - In many parts of the United States, lymphatic mapping and sentinel lymph node biopsy has almost replaced axillary lymph node dissection (ALND) as the axillary staging procedure of choice for small, clinically node-negative breast cancers. However, the long-term outcome of patients undergoing a sentinel lymph node biopsy as the only axillary procedure in patients with tumor-free sentinel nodes remains to be determined. We present the first reported case of axillary recurrence in a patient with breast cancer following a tumor-negative sentinel lymph node biopsy. Whether sentinel lymph node biopsy can replace ALND in the management of patients with early breast cancer remains to be answered. PMID- 12752635 TI - The lactating breast: MRI findings and literature review. AB - Normal physiologic changes in the breast related to pregnancy and lactation can reduce the sensitivity of imaging modalities, such as mammography. This is likely to be true for other breast imaging techniques such as magnetic resonance imaging (MRI). Although malignancy is relatively uncommon in lactating breasts, patients may develop palpable abnormalities that require imaging evaluation. Physiologic changes from pregnancy and lactation can complicate breast imaging. We report the MRI appearance of the lactating breast and address potential difficulties that may be encountered in this clinical situation. PMID- 12752636 TI - Biphenotypic acute leukemia following intensive adjuvant chemotherapy for breast cancer: case report and review of the literature. AB - The risk of secondary leukemia in breast cancer patients who receive adjuvant chemotherapy is an open question. We describe the case a 38-year-old woman who developed acute leukemia 18 months after completion of intense adjuvant chemotherapy with prophylactic granulocyte colony-stimulating factor (G-CSF) support and chest wall irradiation. The diagnosis of biphenotypic T-cell acute myeloid leukemia (AML) was based on morphologic and immunophenotypic criteria. Chromosomal analysis of blasts revealed multiple trisomies and tetrasomies. The patient failed to respond to induction and salvage chemotherapy and died 4 months later. This case of acute leukemia occurred in a cohort of 65 high-risk breast cancer patients who were given intense adjuvant chemotherapy during the last 5 years in our hospital. This is the first case reported in the literature of acute leukemia following intense adjuvant chemotherapy with continuous prophylactic G CSF, which is an actively investigated therapeutic strategy. Vigilance and investigation are needed to determine the leukemogenic potential of intense adjuvant chemotherapy plus radiotherapy in breast cancer patients. A brief review of the literature that deals with acute leukemia that develops after adjuvant chemotherapy for breast cancer and with secondary biphenotypic acute leukemia is presented. PMID- 12752637 TI - Invasive lobular carcinoma in a breast hamartoma. AB - Hamartomas are uncommon benign breast neoplasms. We report the first case, to our knowledge, of invasive lobular carcinoma arising in a breast hamartoma. This case illustrates the importance of careful interpretation of the clinical and mammographic findings. A more aggressive approach toward the management of breast hamartomas is not justified when clinical and mammographic findings are consistent with classic hamartomas. PMID- 12752638 TI - Intracystic papillary carcinoma in the male breast. AB - Intracystic papillary carcinoma is an uncommon noninvasive breast cancer with an excellent prognosis. We report the case of a 74-year-old man. Of the diagnostic tests performed, ultrasonography and pneumocystography were useful for establishing a diagnostic suspicion, however, puncture cytology was inaccurate. Excisional biopsy was necessary to confirm the disease, but also to indicate that local treatment was sufficient. PMID- 12752639 TI - Solitary fibrous tumor of the breast. PMID- 12752640 TI - Angiosarcoma of the breast: mammographic, clinical, and pathologic correlation. PMID- 12752642 TI - Adenosis tumor of the breast. PMID- 12752641 TI - Paget's disease of the male breast: an unusual case of dermal invasion. PMID- 12752643 TI - Interlaboratory variation in results from immunohistochemical assessment of estrogen receptor status. PMID- 12752644 TI - Atypical medullary breast carcinoma in a family carrying the 5382insC BRCA-1 mutation. PMID- 12752645 TI - Review of the current status of tooth whitening with the walking bleach technique. AB - Internal bleaching procedures such as the walking bleach technique can be used for whitening of discoloured root-filled teeth. The walking bleach technique is performed by application of a paste consisting of sodium perborate-(tetrahydrate) and distilled water (3% H2O2), respectively, in the pulp chamber. Following a critical review of the scientific literature, heating of the mixture is contra indicated as the risk of external cervical resorption and the formation of chemical radicals is increased by application of heat. An intracoronal dressing using 30% H2O2 should not be used in order to reduce the risk of inducing cervical resorption. This review provides advice based on the current literature and discusses how the walking bleach technique can lead to successful whitening of non-vital root-filled teeth without the risks of side-effects. PMID- 12752646 TI - Examination of cytotoxicity and mutagenicity of AH26 and AH Plus sealers. AB - AIM: To study in vitro the cytotoxic and mutagenic effects of AH26 and AH Plus. METHODOLOGY: Cytotoxic effects on Chinese hamster V79 cells were determined by counting viable cells following incubation with eluations of AH26 and AH Plus. In one set of experiments, the materials were mixed, set for 1 h and then eluted with dimethyl sulphoxide (DMSO) for 1 h, 24 h and 7 days. In the other set, AH26 and AH Plus were mixed and set for 1 h, 24 h and 7 days in physiological saline then crushed and eluted in DMSO for 24 h. The cytotoxic effects of these eluates were evaluated. Three concentrations were chosen to examine the mutagenic effects of AH26 and AH Plus: 5.57, 16.7 and 55.7 microg mL(-1). The structural chromosomal aberration analysis and micronucleus test were performed on human lymphocytes according to standard procedures. RESULTS: Dose-response curves of cell survival were obtained. Both materials were shown to be cytotoxic in doses larger than 55.7 microg mL(-1), except for AH26, after 7 days setting time. AH Plus was also shown to be toxic in concentrations of 16.7 microg mL(-1), except after 7 days setting time. Neither AH26 nor AH Plus induced a significant increase of chromosomal aberrations or micronuclei induction at any setting time or concentration. CONCLUSION: There was no mutagenicity found for AH26 and AH Plus on human lymphocytes in highly controlled conditions in vitro. PMID- 12752647 TI - Light and scanning electron microscopic evaluation of Glyde File Prep in smear layer removal. AB - AIM: To evaluate the effectiveness of Glyde File Prep used in conjunction with sodium hypochlorite irrigation in the removal of smear layer produced during root canal instrumentation. METHODOLOGY: Thirty-nine extracted human teeth with single root canals were used. Access cavities were prepared and the teeth divided into three groups of 13 teeth each. Each group was treated by one of the three different regimes of irrigation and conditioning during root canal instrumentation. Group A: 0.5 mL of 1% NaOCl irrigation after each file size with an additional final irrigation of 10 mL 1% NaOCl; group B: 0.5 mL of 1% NaOCl irrigation after each file size with an additional final irrigation of 10 mL 17% EDTA; group C: Glyde File Prep coated on each instrumentation file used in conjunction with 0.5 mL 1% NaOCl irrigation after each file size and an additional final irrigation of 10 mL 1% NaOCl. The teeth were then longitudinally grooved and sectioned. Root canal cleanliness was evaluated with the aid of a Nikon light microscope (x40 and x100) and scanning electron microscope (x1000 and x3000). The debris scores obtained at three canal regions were compared statistically within the same group and among different groups using repeated measurements of analysis of variance (anova) with Bonferroni adjustments and anova with posthoc Tukey HSD, respectively. RESULTS: The canals treated with EDTA and Glyde File Prep were significantly cleaner than those treated with NaOCl alone. The apical region of the root canals generally displayed more residual smear layer, but the difference was not significant. CONCLUSIONS: Used in conjunction with NaOCl irrigation, Glyde File Prep was effective in removing smear layer produced during root canal instrumentation. PMID- 12752648 TI - Endodontic treatment performed by Flemish dentists. Part 2. Canal filling and decision making for referrals and treatment of apical periodontitis. AB - AIM: To gather information on root-canal treatment carried out by dentists working in Flanders (Belgium). METHODOLOGY: A questionnaire reported in a previous study was also used to gather information on canal medicaments, canal filling, and in decision making for referrals and treatment of apical periodontitis. RESULTS: Calcium hydroxide as an interappointment dressing was used by 69.7% of the respondents. Approximately one-third of the respondents did not use any intracanal medicament. Caustic products used for pulp tissue fixation were used by 66.8% of the respondents. Cavit (48.2%) and glass-ionomer (31.3%) were the temporary coronal-filling materials used most often, followed by zinc oxide-eugenol and IRM(R). Cold lateral condensation of gutta-percha was the filling technique most used by the respondents (65.8%). Single-cone gutta-percha placement (16%), paste techniques (4.9%) and silver points (3.9%) were still used. Resin-based sealers were used most often (88.6%). Paraformaldehyde containing sealers such as Endomethasone and N2 were used infrequently. Approximately half of the practitioners were satisfied with their canal-filling technique, others felt that they could do better (43.0%); 0.7% were not satisfied. In cases with apical periodontitis, the size of periapical lesions and/or the presence of a root filling influenced the choice of endodontic treatment. The most common reasons for referral of endodontic cases were: retrieval of silver points, surgery, and post removal. CONCLUSIONS: The results of this study indicate that techniques and methods used for canal medication and canal filling were acceptable for the majority of the respondents. Re-treatment was underestimated as a treatment option. PMID- 12752649 TI - Induction of interleukin-6 gene expression by pro-inflammatory cytokines and black-pigmented Bacteroides in human pulp cell cultures. AB - AIM: To investigate the effect of pro-inflammatory cytokines and black-pigmented Bacteroides on the expression of IL-6 gene in human pulp fibroblasts. METHODOLOGY: IL-1alpha, tumour necrosis factor-alpha (TNF-alpha) and the supernatants of Porphyromonas endodontalis, P. gingivalis and Prevotella intermedia were used to evaluate IL-6 gene expression in human pulp fibroblasts. The levels of mRNAs were measured by the quantitative reverse-transcriptase polymerase chain reaction analysis. RESULTS: Investigations of the time dependence of IL-6 mRNA expression in pro-inflammatory cytokines-treated cells revealed a rapid accumulation of the transcript after 2 h of exposure and remained elevated throughout the 24-h incubation period. In addition, black pigmented Bacteroides also induced IL-6 gene expression in human pulp fibroblasts. CONCLUSIONS: Pro-inflammatory cytokines and black-pigmented Bacteroides may be involved in developing pulpal inflammation through the stimulation of IL-6 production. PMID- 12752650 TI - A comparative study of root canal preparation using FlexMaster and HERO 642 rotary Ni-Ti instruments. AB - AIM: To compare several parameters of root canal preparation using two different rotary nickel-titanium (Ni-Ti) instruments: FlexMaster (VDW, Munich, Germany) and HERO 642 (Micro-Mega, Besancon, France). METHODOLOGY: Fifty extracted human mandibular molars with root canal curvatures between 20 and 40 degrees were embedded into a muffle system. All root canals were prepared to size 45 using a high-torque motor with two different Ni-Ti instruments, FlexMaster and HERO 642. In both groups, irrigation was performed with 2 mL NaOCl (3%) after each instrument size. RC-Prep (Premier, Philadelphia, USA) was used as a chelating agent with each instrument. The following parameters were evaluated: straightening of curved root canals, postoperative root canal diameter, working safety (file fractures, perforations, apical blockages, loss of working length), cleaning ability and working time. Statistical analysis was performed using the Wilcoxon's test (P < 0.05) for straightening and Fishers's exact test (P < 0.05) for comparison of cross-sections, contact between pre- and postoperative diameter, root canal cleanliness and working time. RESULTS: Both Ni-Ti systems maintained the curvature well: the mean degree of straightening was 0.6 degrees for FlexMaster and 0.5 degrees for HERO 642. One file was fractured with the FlexMaster system, but further procedural incidents were not recorded. Following preparation with FlexMaster, 18% of the root canals had a round diameter, 53% an oval diameter and 29% an irregular diameter; HERO 642 preparations resulted in a round diameter in 25%, oval shape in 47% and irregular cross-sections in 28% of the cases. Mean working time was shorter for HERO 642 (66.0 s) than for FlexMaster (71.1 s). Cleanliness of the root canal walls was investigated under the SEM using 5-score indices for debris and smear layer. For debris, HERO 642 and FlexMaster achieved 73 and 70% scores of 1 and 2, respectively. The results for smear layer were similar: HERO 642 and FlexMaster achieved 33 and 26% scores of 1 and 2, respectively. Significant differences between the two systems were not detected for any of the parameters evaluated. CONCLUSIONS: Both systems respected original root canal curvature well and were safe. Both systems failed to remove debris and smear layer in the majority of the cases. PMID- 12752651 TI - In vitro evaluation of apical microleakage following canal filling with a coated carrier system compared with lateral and thermomechanical Gutta-Percha condensation techniques. AB - AIM: The apical sealing ability of a coated carrier system was evaluated in extracted human teeth and compared with lateral and thermomechanical condensation techniques using dye penetration. METHODOLOGY: Sixty-four extracted single-rooted teeth were instrumented to an apical size 30 using 4% taper Hero 642 trade mark instruments (Micromega, Besancon, France). The sample was divided into three experimental groups. Twenty teeth were randomly obturated with lateral condensation, 20 with thermomechanical compaction and 20 teeth with the Herofill trade mark Soft-Core system. Four teeth were used as positive and negative controls. The teeth were covered with nail varnish up to 2 mm from the apical foramen and immersed in a 2% aqueous methylene blue dye solution for 1 week and then washed, dehydrated and embedded in resin. The apical 1 mm of each tooth was removed to reveal the apical limit of the preparation. Transverse sections of the teeth were taken at 500, 1000 and 1500 micro m from this point and evaluated for apical leakage. Significant differences between the preparations were analysed with a semiautomatic analyser and the ratio of the dye-penetrated surface to the total dentinal surface was calculated. RESULTS: Statistical analysis of the results demonstrated significantly less leakage for the Herofill trade mark Soft Core system compared to lateral condensation in terms of total mean dentinal surface and at the 500 micro m level. No other differences were noted between Herofill trade mark Soft-Core and thermomechanical or lateral condensation, either for the total mean value or at each level. CONCLUSIONS: The Herofill trade mark Soft-Core system was a reliable obturation system in the apical portion and compared favourably with other gutta-percha filling techniques. PMID- 12752652 TI - Endodontic treatment of developmental anomalies in posterior teeth: treatment of geminated/fused teeth--report of two cases. AB - AIM: Gemination or fusion is a rare occurrence in the mandibular posterior teeth. Endodontic treatment of these teeth needs special care and attention to the bizarre anatomy. The aim of this article is to describe the problems encountered and the strategy in treating such cases. CASE REPORT: Two cases of complex endodontic treatment of fused/geminated teeth are presented. The first is an 11 year-old girl with an anomalous 'double' first mandibular molar and premolar diagnosed as having necrotic pulp with chronic apical abscess of endodontic origin; the second is a 16-year-old boy with 'double' second and supernumerary mandibular molars, who was diagnosed with irreversible pulpitis. Both cases were treated successfully in multiple appointments. The common features and treatment modalities are discussed. KEY LEARNING POINTS: Failure to diagnose fused/geminated teeth leads to misdiagnosis and a treatment plan that could cause permanent damage and tooth loss. Generally, there is communication between root canal systems of fused/geminated teeth which should be treated as one entity. Use of magnification is an important aid during treatment. PMID- 12752653 TI - Transposable element (TE) display and rapid detection of TE insertion polymorphism in the Anopheles gambiae species complex. AB - Transposable element (TE) display was shown to be a highly specific and reproducible method of detecting the insertion sites of TEs in individuals of the African malaria mosquito, Anopheles gambiae, and its sibling species, A. arabiensis. Relatively high levels of insertion polymorphism were observed during the TE display of several families of miniature inverted-repeat TEs (MITEs) that have variable copy numbers. The genomic locations of selected insertion sites were identified by matching the sequences of their corresponding bands in a TE display gel to specific regions of the draft A. gambiae genome assembly. We discuss different scenarios in which TE display will provide powerful dominant and co-dominant genetic markers to study the behaviour of TEs in A. gambiae populations and to illustrate the complex population genetics of this intriguing disease vector. We suggest that TE display can also provide tools for a phylogenetic analysis of the A. gambiae complex. PMID- 12752654 TI - B96Bom encodes a Bombyx mori tyramine receptor negatively coupled to adenylate cyclase. AB - A cDNA encoding a biogenic amine receptor (B96Bom) was isolated from silkworm (Bombyx mori) larvae, and the ligand response of the receptor stably expressed in HEK-293 cells was examined. Tyramine (TA) at 0.1-100 micro m reduced forskolin (10 micro m)-stimulated intracellular cAMP levels by approximately 40%. The inhibitory effect of TA at 1 micro m was abolished by yohimbine and chlorpromazine (each 10 micro m). Although octopamine (OA) also reduced the cAMP levels, the potency was at least two orders of magnitude lower than that of TA. Furthermore, unlabelled TA (IC50 = 5.2 nm) inhibited specific [3H]TA binding to the membranes of B96Bom-transfected HEK-293 cells more potently than did OA (IC50 = 1.4 micro m) and dopamine (IC50 = 1.7 micro m). Taken together with the result of phylogenetic analysis, these findings indicate that the B96Bom receptor is a B. mori TA receptor, which is negatively coupled to adenylate cyclase. The use of this expression system should facilitate physiological studies of TA receptors as well as structure-activity studies of TA receptor ligands. PMID- 12752655 TI - Molecular analysis of the serine/threonine kinase Akt and its expression in the mosquito Aedes aegypti. AB - A key component of the insulin-signalling pathway, the protein kinase Akt, was identified and cloned as a cDNA from ovaries of the mosquito Aedes aegypti. An ortholog gene was found in the Anopheles gambiae genome database, and like other Akts, both mosquito Akts possess pleckstrin homology domains for membrane binding and a serine/threonine kinase domain. When Ae. aegypti ovaries were treated with bovine insulin in vitro, a putative Akt was threonine-phosphorylated, as expected for Akts. AaegAKT was only expressed in embryos for the first 6 h after oviposition and in ovaries before and during a gonotrophic cycle. PMID- 12752656 TI - An endothelin-converting enzyme homologue in the locust, Locusta migratoria: functional activity, molecular cloning and tissue distribution. AB - Endothelin-converting enzyme is the key enzyme in the process of endothelin production. Endothelin is a peptide that plays an important role in vasoconstriction and the development of neural crest-derived cells in vertebrates. Activity assays performed on membrane extracts from Locusta migratoria brain revealed the existence of a protease activity responsible for the formation of mature endothelin-1 from its precursor, big endothelin. Cloning experiments led to a cDNA sequence (Lom ECE) with an open reading frame of 727 amino acid residues displaying all the characteristic ECE features. A comparison of ECE activity levels among different tissues of the locust showed a high enzyme activity in the gonads and midgut. RT-PCR experiments showed a wide tissue distribution of Lom ECE mRNA, with transcription being most abundant in brain tissue. PMID- 12752657 TI - The pyrethroid knock-down resistance gene in the Anopheles gambiae complex in Mali and further indication of incipient speciation within An. gambiae s.s. AB - In Mali the Anopheles gambiae complex consists of An. arabiensis and Mopti, Savanna and Bamako chromosomal forms of An. gambiae s.s. Previous chromosomal data suggests a complete reproductive isolation among these forms. Sequence analysis of rDNA regions led to the characterization of two molecular forms of An. gambiae, named M-form and S-form, which in Mali correspond to Mopti and to Savanna/Bamako, respectively, while it has failed so far to show any molecular difference between Savanna and Bamako. The population structure of An. gambiae s.l. was analysed in three villages in the Bamako and Sikasso areas of Mali and the frequency of pyrethroid resistance of the knock-down resistance (kdr) type was calculated. The results show that the kdr allele is associated only with the Savanna form populations and absent in sympatric and synchronous populations of Bamako, Mopti and An. arabiensis. This is the first molecular indication of barriers to gene flow between the Bamako and Savanna chromosomal forms. Moreover, analyses of specimens collected in the Bamako area in 1987 show that the kdr allele was already present in the Savanna population at that time, and that the frequency of this allele has gradually increased since then. PMID- 12752658 TI - Identification of regulatory regions in the DNA puff BhC4-1 promoter. AB - The mechanisms that control DNA puff BhC4-1 expression in the salivary gland of sciarid late larvae have been shown to be conserved in Drosophila. By analysing Drosophila transformed with constructs carrying progressive deletions of the BhC4 1 promoter fragment (-3314/+40) fused to the lacZ reporter gene we show that the elements required for the correct BhC4-1-lacZ developmental regulation in prepupal salivary glands are contained in a 226 bp fragment (-186/+40). Also, interestingly, this study identified a 67 bp fragment (-253/-187) that activates BhC4-1-lacZ expression specifically in the ring gland. PMID- 12752659 TI - Identification and localization of the nicotinic acetylcholine receptor alpha3 mRNA in the brain of the honeybee, Apis mellifera. AB - The nicotinic acetylcholine receptors are ligand-gated ion channels responsible for rapid neurotransmission and are target sites for pesticides in insects. In the honeybee Apis mellifera, pharmacological and electrophysiological studies have shown that different nicotinic acetylcholine receptor subtypes may exist in the brain. Here, we have identified a honeybee cDNA that encodes a 537 amino acid protein with features typical of nicotinic acetylcholine receptor alpha subunit, and sequence homology to human alpha3. In situ hybridization on cryosections shows that the Apisalpha3 mRNA is differently expressed in larvae and adult. In larvae, Apisalpha3 mRNA expression is restricted to the suboesophageal ganglia. In adult, it is further expressed in the optic lobes, the dorsal lobes, the antennal lobes and the calyces of mushroom bodies. Together our results suggest that Apisalpha3 shows a controlled expression pattern during development. PMID- 12752660 TI - The scarlet eye colour gene of the tephritid fruit fly: Bactrocera tryoni and the nature of two eye colour mutations. AB - A homologue of the Drosophila melanogaster eye-colour gene, scarlet (st), has been isolated from the genome of the tephritid fruit fly, Bactrocera tryoni. The comparison of the B. tryoni and D. melanogaster scarlet gene shows 71.2% and 79.3% sequence identity at the DNA and the derived amino acid level, respectively. Two allelic eye-colour mutations of B. tryoni, orange-eyes and lemon-eyes, have been recovered and found to be colocalized with the st gene. The st gene sequence in the two mutant strains has been examined for DNA sequence changes and expression levels. PMID- 12752661 TI - Quantitative expression analysis of a Glyptapanteles indiensis polydnavirus protein tyrosine phosphatase gene in its natural lepidopteran host, Lymantria dispar. AB - In the present study, expression of a newly identified Glyptapanteles indiensis polydnavirus (GiPDV) gene encoding a putative protein tyrosine phosphatase (PDVPTP) was monitored in vivo in the parasitized host, L. dispar, using one step RT-PCR. Expression levels of the PDVPTP transcript were also evaluated in various host tissues at different times post parasitization (pp) using RT quantitative competitive PCR (RT-qcPCR). Expression levels varied, with the most abundant transcript detected in host haemolymph 2 h pp. The high expression level in host haemolymph at an early stage of parasitization suggested a potential role for viral PDVPTP in disruption of the host immune system and protection of the endoparasitoid egg from encapsulation. Additionally, the PDVPTP gene or its homolog(s) mapped to more than one GiPDV genomic DNA segment, which may account for its increased level of expression in the absence of virus replication. PMID- 12752662 TI - Variations in mitochondrial DNA and gene transcription in freezing-tolerant larvae of Eurosta solidaginis (Diptera: Tephritidae) and Gynaephora groenlandica (Lepidoptera: Lymantriidae). AB - Respiration, mitochondrial (mt)DNA content, and mitochondrial-specific RNA expression in fat body cells from active and cold-adapted larvae of the goldenrod gall fly, Eurosta solidaginis, and the Arctic woolly bear caterpillar, Gynaephora groenlandica, were compared. Reduced amounts of mtDNA were observed in cold adapted larvae of both E. solidaginis and G. groenlandica collected in fall or winter, compared with summer-collected larvae. mtDNA increased to levels similar to those of summer-collected larvae after incubation at 10 degrees C or 15 degrees C for 5 h. Mitochondrial-specific RNAs (COI and 16S) were observed in fat body cells of both active and cold-adapted E. solidaginis larvae. Our results suggest that mitochondrial proteins required for respiration may be restored rapidly from stable RNAs present in overwintering larvae. PMID- 12752663 TI - Identification of a tachykinin-related neuropeptide from the honeybee brain using direct MALDI-TOF MS and its gene expression in worker, queen and drone heads. AB - Using a combination of MALDI-TOF and on-line capillary HPLC/Q-Tof mass spectroscopy, we identified and determined the amino acid sequence of a novel neuropeptide in the brain of the honeybee Apis mellifera L., termed AmTRP peptide (Apis mellifera tachykinin-related peptide), related to insect tachykinin. A cDNA for a prepro-protein (prepro-AmTRP) of AmTRP was isolated and determined to encode seven AmTRPs 1-7. Northern blot analysis indicated that the prepro-AmTRP gene is expressed differentially in the nurse bee, forager, queen and drone heads. Strong expression was detected in the queen and forager heads, while weak and almost no significant expression was detected in the nurse and drone heads, respectively. These results suggest that AmTRP peptide functions as a neuromodulator and/or hormone, associated with sex-specific or age/division of labour-selective behaviour and/or physiology of the honeybees. PMID- 12752664 TI - RNA interference in ticks: a study using histamine binding protein dsRNA in the female tick Amblyomma americanum. AB - RNA interference (RNAi), a gene silencing process, has been recently exploited to determine gene function by degrading specific mRNAs in several eukaryotic organisms. We constructed a double stranded RNA (dsRNA) from a previously cloned putative Amblyomma americanum histamine binding protein (HBP) to test the significance of using this methodology in the assessment of the function and importance of gene products in ectoparasitic ticks. The female salivary glands incubated in vitro with HBP dsRNA had a significantly lower histamine binding ability. In addition, the injection of HBP dsRNA into the unfed females led both to a reduced histamine binding ability in the isolated salivary glands and to an aberrant tick feeding pattern or host response. Molecular data demonstrated less expression of the HBP mRNA in the RNAi group. Taken together, these results suggest that RNAi might be an important tool for assessing the significance of tick salivary gland secreted proteins modulating responses at the tick-host interface. PMID- 12752665 TI - Overview: apoptotic signaling pathways in the immune system. PMID- 12752666 TI - Caspase-activation pathways in apoptosis and immunity. AB - Members of the caspase family of cysteine proteases have been firmly established to play key roles in signal transduction cascades that culminate in apoptosis (programmed cell death). Caspases are normally expressed as inactive precursor enzymes (zymogens) that become activated during apoptosis and proceed to dismantle the cell from within. To date, three major apoptosis-associated pathways to caspase activation have been elucidated. Certain caspases, such as caspase-1, also occupy important positions in signaling pathways associated with immune responses to microbial pathogens. In this situation, caspase activation is associated with the maturation of pro-inflammatory cytokines, such as interleukin 1beta (IL-1beta) and IL-18, and not apoptosis per se. Here, we discuss the current understanding of how caspases are activated during apoptosis and inflammation and the roles these proteases play in either context. PMID- 12752667 TI - Caspases and T lymphocytes: a flip of the coin? AB - In this review, we consider the role caspases play in cell death downstream of death receptors and cell intrinsic death mechanisms. In particular, we focus on these mechanisms in antigen-induced cell death, a mechanism which regulates the number of surviving T cells at the end of an immune response. The relative role of the apoptosome as an amplifier rather than an initiator of apoptosis is considered. Several factors that regulate the susceptibility to activation induced cell death are considered. These factors emanate from the stimulation of the T-cell receptors and include multiple pathways. Recent work has shown that death receptor signaling can play an interesting role in cell proliferation in both humans and animals. These recent findings are discussed in the light of models of death receptor signaling. PMID- 12752668 TI - Granzyme B: a natural born killer. AB - A main pathway used by cytotoxic T lymphocytes (CTLs) and natural killer cells to eliminate pathogenic cells is via exocytosis of granule components in the direction of the target cell, delivering a lethal hit of cytolytic molecules. Amongst these, granzyme B and perforin have been shown to induce CTL-mediated target cell DNA fragmentation and apoptosis. Once released from the CTL, granzyme B binds its receptor, the mannose-6-phosphate/insulin-like growth factor II receptor, and is endocytosed but remains arrested in endocytic vesicles until released by perforin. Once in the cytosol, granzyme B targets caspase-3 directly or indirectly through the mitochondria, initiating the caspase cascade to DNA fragmentation and apoptosis. Caspase activity is required for apoptosis to occur; however, in the absence of caspase activity, granzyme B can still initiate mitochondrial events via the cleavage of Bid. Recent work shows that granzyme B mediated release of apoptotic factors from the mitochondria is essential for the full activation of caspase-3. Thus, granzyme B acts at multiple points to initiate the death of the offending cell. Studies of the granzyme B death receptor and internal signaling pathways may lead to critical advances in cell transplantation and cancer therapy. PMID- 12752669 TI - Regulation of apoptosis by ubiquitination. AB - Cell elimination through apoptosis, or programmed cell death, is an evolutionarily conserved central tenet of biology from embryological development to immune homeostasis. While many of the apoptotic signaling pathways have been elucidated, the relationship between ubiquitin and apoptosis is only beginning to be defined. In the past decade, many reports of polyubiquitin conjugation of key pro- and anti-apoptotic molecules have characterized ubiquitin as an essential regulatory modification targeting proteins for proteasomal degradation. However, recent work relating monoubiquitination and nonclassical polyubiquitin conjugation to apoptotic molecules has added an additional level of diversity to the role of ubiquitin in apoptotic regulation beyond degradation. This review focuses on the direct effects of ubiquitination on apoptosis-signaling molecules. PMID- 12752670 TI - Death and Baxes: mechanisms of lymphotrophic cytokines. AB - In this review, we briefly cover the critical requirements for interleukin-7 (IL 7) in thymocyte development and peripheral T-cell homeostasis. Part of the IL-7 effect is antiapoptotic or 'trophic' and we have studied the intracellular pathways involved in lymphocyte survival and death regulated by this cytokine. We review the evidence for a role of the JAK signal transducers and activators of transcription protein (STAT) pathway and phosphoinositide 3-kinase (PI3K)-AKT pathways in survival. The death pathway following IL-7 withdrawal is discussed in terms of the balance of BCL-2 vs. BAX and other death proteins and the role of metabolic disturbances involving glucose metabolism and intracellular pH. The IL 7 survival and death pathways in lymphocytes may be representative of many trophic factors in different cell types; yet we conclude that much of the mechanism remains to be discovered. PMID- 12752671 TI - The role of CD95 in the regulation of peripheral T-cell apoptosis. AB - Apoptosis of activated peripheral T cells during the termination phase of an immune response is critical to maintain T-cell homeostasis. Activated T cells can be removed by two mechanisms: activation-induced cell death (AICD) and death by neglect. AICD is triggered by death receptors, whereas death by neglect is induced by cytokine withdrawal. CD95 (APO-1/Fas) belongs to the subfamily of death receptors and plays a major role in AICD. In this review, we focus on the molecular mechanisms of AICD, in particular those involving the CD95 system. Moreover, we discuss the relative contribution of AICD and death by neglect to terminate a T-cell immune response. In order to become fully activated, T cells require a second signal provided by antigen-presenting cells. We discuss how these costimulatory signals counteract pro-apoptotic signals and, finally, which signals might protect T cells from death to generate a pool of memory T cells. PMID- 12752672 TI - Activation-induced cell death in T cells. AB - A properly functioning immune system is dependent on programmed cell death at virtually every stage of lymphocyte development and activity. This review addresses the phenomenon of activation-induced cell death (AICD) in T lymphocytes, in which activation through the T-cell receptor results in apoptosis. AICD can occur in a cell-autonomous manner and is influenced by the nature of the initial T-cell activation events. It plays essential roles in both central and peripheral deletion events involved in tolerance and homeostasis, although it is likely that different forms of AICD proceed via different mechanisms. For example, while AICD in peripheral T cells is often caused by the induction of expression of the death ligand, Fas ligand (CD95 ligand, FasL), it does not appear to be involved in AICD in thymocytes. This and other mechanisms of AICD are discussed. One emerging model that may complement other forms of AICD involves the inducible expression of FasL by nonlymphoid tissues in response to activated T lymphocytes. Induction of nonlymphoid FasL in this manner may serve as a sensing mechanism for immune cell infiltration, which contributes to peripheral deletion. PMID- 12752673 TI - The control of apoptosis in lymphocyte selection. AB - The stochastic nature of rearrangement and diversification of the gene segments encoding immunoglobulins (Igs) and T cell receptors (TCRs) inevitably gives rise to immature B and T lymphocytes that lack antigen receptors or express useless or dangerous (self-antigen-specific) ones. Signaling through antigen receptors promotes survival, proliferative expansion and further differentiation of useful cells and deletion of the useless and dangerous ones. During immune responses, pathogen-specific B and T lymphocytes, as well as cells of the innate immune system, undergo extensive proliferation and develop effector functions, such as antibody secretion, cytotoxicity or cytokine production. To prevent tissue damage by these effector molecules, activated lymphocytes are removed when an infection has been overcome. Together with other mechanisms, including developmental arrest and induction of unresponsiveness (anergy), programmed cell death (apoptosis) of autoreactive lymphocytes safeguards immunological tolerance to self and assists in the development of an effective immune system. We have been investigating the molecular mechanisms that control programmed cell death. This review describes some of our experiments using transgenic and knockout mice, which overexpress or lack apoptosis regulators, that led to discoveries on how life and death decisions are made during development and functioning of the immune system. PMID- 12752674 TI - Transducing signals from antigen receptors to nuclear factor kappaB. AB - Signaling leading to the survival or apoptosis of immune system cells must be balanced to ensure the normal mounting and extinguishing of immune responses. One of the essential regulators of immune cell survival is the transcription factor nuclear factor kappaB (NF-kappaB). NF-kappaB is critical for the activation of T and B lymphocytes and is a central coordinator of innate and adaptive immunity. Pathogen recognition, whether mediated via the Toll-like receptors or via the antigen-specific T- and B-cell receptors, initiates the activation of distinct signal transduction pathways that activate NF-kappaB. Activation of NF-kappaB by these pathways is necessary for lymphocyte activation, expansion, and effector function in response to infection. In addition, recent work has shown that the aberrant activation of NF-kappaB by these pathways can contribute to the development of autoimmunity, chronic inflammation, or lymphoid malignancy. There is thus an urgent need to understand the exact molecular details of these signal transduction cascades so that we may develop novel therapeutics. This article will review the specific signal transduction pathways that mediate NF-kappaB activation in response to antigen receptor ligation in T and B lymphocytes. These newly defined pathways, which are essential for adaptive immune responses, are built around the key adapter protein, Bcl-10. Bcl-10 is known to participate in chromosomal translocations in human mucosa-associated lymphoid tissue lymphomas. PMID- 12752675 TI - Neutrophil apoptosis pathways and their modifications in inflammation. AB - Neutrophils are constantly produced in large numbers in the bone marrow, and the same numbers of cells need to die within a defined time period in order to keep cellular homeostasis under physiologic conditions. Changing the rate of apoptosis rapidly changes cell numbers in such systems. For instance, in many bacterial and autoimmune inflammatory diseases, delayed apoptosis is one important mechanism for neutrophil accumulation. Excessive production of granulocyte/macrophage colony-stimulating factor (GM-CSF) and granulocyte colony-stimulating factor (G CSF), two important neutrophil survival factors, is often observed in such inflammatory responses. Cytokine withdrawal, as it occurs in the resolution phase of inflammation, leads to the induction of apoptosis. Moreover, neutrophil apoptosis can be accelerated both in the presence and in the absence of survival factors by activation of distinct members of the tumor necrosis factor/nerve growth factor receptor family. This review focuses on recently published work regarding signaling pathways that regulate neutrophil apoptosis. PMID- 12752676 TI - Apoptosis, tolerance, and regulatory T cells--old wine, new wineskins. AB - Antigen-specific unresponsiveness (or tolerance) has always been an important area of research. Interest in the fate of apoptotic cells and their ability to tolerize has revived interest in some of the older models involving hapten modified self. Recently, we have examined the mechanisms by which intravenous injection of trinitrophenol-coupled spleen cells leads to systemic tolerance. These studies have revealed an important role for Fas/Fas ligand interactions, caspases, CD40/CD40L, and regulatory CD4+ and CD8+ T cells. Extension of these studies to peripheral deletion of T-cell antigen receptor transgenic T cells has shown that deletion and active regulation of immune responses may be important mechanisms for the control of potentially damaging autoimmune responses. PMID- 12752677 TI - Apoptosis and transplantation tolerance. AB - Self-tolerance is maintained by several mechanisms including deletion (via apoptosis) and regulation. Acquired tolerance to allogeneic tissues and organs exploits similar strategies. One key difference between alloantigens and peptide antigens is the enormous number of T cells that are alloreactive. Accumulating evidence suggests that in the face of this large mass of potentially graft destructive T cells, tolerance requires an initial wave of deletion. This creates a more level playing field in which a smaller number of regulatory T cells can then act to maintain an established tolerant state. Deletion of alloreactive T cells by apoptosis actively promotes immunoregulation as well, by interfering with proinflammatory maturation of antigen presenting cells. This article reviews the immune response to alloantigens, the development and use of both necrotic and apoptotic means of cell death during the evolution of the immune response, and the likely role and mechanisms by which apoptosis promotes, and may even be required for, transplantation tolerance. PMID- 12752679 TI - Isolation of antigen-specific B cells. AB - Cell separation techniques are important in immunology. Major cell populations can be separated successfully with high purity. However, isolation of cells which are specific for particular antigens is more challenging because of the relatively small numbers of antigen-specific cells, and the lack of independent markers available to determine the purity of the isolated population. In this review, the literature describing three principal techniques used to separate antigen-specific cells has been reviewed. Particular emphasis has been placed on yield and purity; the two most important parameters of any purification method. The most promising isolation methods have used immunomagnetic sorting and multiparametric flow cytometric analysis. PMID- 12752680 TI - Ikaros, Aiolos and Helios: transcription regulators and lymphoid malignancies. AB - Ikaros, Aiolos and Helios encode zinc finger transcription factors that are important regulators of lymphoid development and differentiation. These proteins are involved in the control of gene expression and when associated with nuclear complexes, participate in nucleosome remodeling. Because differential splicing produces multiple protein isoforms with potentially different functions, the Ikaros protein family provides a useful model for the study of whether post transcriptional modifications are involved in tumoral transformation. Several reports reinforce the hypothesis that Ikaros, Aiolos and Helios expression is deregulated in human leukaemias. The direct involvement of aberrant protein expression of Ikaros family members in human haematological malignancies is discussed. PMID- 12752681 TI - Immunity in the absence of CD28 and CD137 (4-1BB) molecules. AB - We report the generation and immune regulation of mice that are deficient in CD28 and 4-1BB (CD137) genes. These mice were viable, fertile and did not display any overt abnormalities and had a normal T cell phenotype in thymus and spleen. Proliferative responses to anti-CD3 and ConA were enhanced in 4-1BB-/- but not in either CD28-/- or double mutant mice, while levels of interleukin-2 were decreased in all mutant mice. Although the 4-1BB-/- mice displayed increased basal levels of most immunoglobulin isotypes tested, the plateau levels of immunoglobulin G2a, immunoglobulin G2b and immunoglobulin A were particularly high compared to wild type controls. The immunoglobulin class switch to T dependent antigen was normal in 4-1BB-/- mice but was greatly affected in both CD28-/- and 4-1BB-/- CD28-/- mice. Vesicular stomatitis virus-specific cytotoxic T lymphocyte responses and plaque reduction neutralizing ability was differentially reduced in all mutant mice. Contact sensitivity to allergens showed marginal but not significant change in ear thickness in 4-1BB-/- mice, but an ability to mount contact hypersensitivity to the same antigens was greatly curtailed in CD28-/- and double mutant mice. PMID- 12752682 TI - A prolonged immune response to antigen delivered in poly (epsilon-caprolactone) microparticles. AB - A single dose vaccine formulation which induces both humoral and cell-mediated immune responses over a prolonged period would provide a potent weapon against infectious disease. We have used a water-in-oil-in-oil, solvent evaporation method for generating poly epsilon-caprolactone microparticles and tested their ability to induce an immune response against the model antigen ovalbumin. We hypothesized that the initial release of antigen from the surface of the poly epsilon-caprolactone microparticles would act as the priming dose and that the delayed release over the following months, due to diffusion from or break-down of the microparticles, would act as a boost to the immune response. Ovalbumin encapsulated in the poly epsilon-caprolactone microparticles was able to induce both antibody and cell-mediated immune responses. However our results suggest that the spontaneous release had little effect on the immune response. Despite this the response was maintained for at least 8 months following a single immunization. Both humoral and cell-mediated immune responses were induced in mice. This simple method of vaccine formulation offers a cost-efficient way to deliver antigen in a single dose to the immune system. PMID- 12752683 TI - FcgammaRIIA polymorphisms in Streptococcus pneumoniae infection. AB - Invasive pneumococcal disease continues to be a major cause of morbidity and mortality among children and adults worldwide. Effective host defence against Streptococcus pneumoniae depends on immunoglobulin G-mediated phagocytosis of the bacteria and it has been shown in vitro that the FcgammaRIIA polymorphism (FcgammaRIIA-R131 vs FcgammaRIIA-H131) determines the capacity of immunoglobulin G2-mediated phagocytosis via this receptor. In this study, we evaluated FcgammaRIIA polymorphisms in children with pneumococcal sepsis and a number of control groups in order to investigate a possible association of FcgammaRIIA genotypes with Streptococcus pneumoniae infection. The distribution of the genotypes differed in these populations. The frequency of homozygosity for FcgammaRIIA-R/R131 in the patients was significantly higher than that in the healthy random donor population (43%vs 21%, P < 0.05). The frequencies of FcgammaRIIA-H/H131 were similar among all groups of individuals, while the incidence of the heterozygous FcgammaRIIA-R/H131 was lower (35%vs 52%, P < 0.05). Thus, it appears that the FcgammaRIIA-H131 polymorphic form, even in the heterozygous form, may be protective for pneumococcal sepsis and children with FcgammaRIIA-R/R131 genotype could be more at risk of infection with invasive Streptococcus pneu-moniae. PMID- 12752684 TI - Evasion of innate and adaptive immunity by flaviviruses. AB - After a virus infects an animal, antiviral responses are generated that attempt to prevent dissemination. Interferons, antibody, complement, T and natural killer cells all contribute to the control and eradication of viral infections. Most flaviviruses, with the exception of some of the encephalitic viruses, cause acute disease and do not establish persistent infection. The outcome of flavivirus infection in an animal is determined by a balance between the speed of viral replication and spread, and the immune system response. Although many of the mechanistic details require further elucidation, flaviviruses have evolved specific tactics to evade the innate and adaptive immune response. A more thorough understanding of these principles could lead to improved models for viral pathogenesis and to strategies for the development of novel antiviral agents. PMID- 12752685 TI - Interaction of flaviviruses with cells of the vertebrate host and decoy of the immune response. AB - Flaviviruses cause endemic and epidemic disease with significant morbidity and mortality throughout the world. In contrast to viruses that avoid the host immune response by down-regulating cell surface major histocompatibility complex expression, infection by members of the neurotropic Japanese encephalitis serogroup induce virus-directed functional increases in expression of class I and II major histocompatibility complex and various adhesion molecules, resulting in increased susceptibility to both virus- and major histocompatibility complex specific cytotoxic T lymphocyte lysis. These changes are comodulated by T1 and T2 cytokines, as well as by cell cycle position and adherence status at infection. Infected skin dendritic (Langerhans) cells also show increased costimulatory molecule expression and local interleukin-1beta production causes accelerated migration of Langerhans cells to local draining lymph nodes, where initiation of antiviral immune responses occur. The exact mechanism(s) of up-regulation is unclear, but changes are associated with NF-kappaB activation and increased MHC and ICAM-1 gene transcription, independently of interferon or other pro inflammatory cytokines. We hypothesize that these viruses may decoy the adaptive immune system into generating low-affinity, self-reactive T cells which clear virus poorly, as part of their survival strategy. This may enable viral growth and immune escape in cycling cells, which do not significantly up-regulate cell surface molecules. A possible side-effect of this might be immunopathology, caused by 'autoimmune' cross-reactive damage of uninfected high major histocompatibility complex and adhesion molecule-expressing cells, with consequent exacerbation of encephalitic disease. Results from a murine model of flavivirus encephalitis developed in this laboratory further suggest that interferon-gamma plays a crucial role in fatal immunopathology. PMID- 12752686 TI - MHC class I up-regulation by flaviviruses: Immune interaction with unknown advantage to host or pathogen. AB - In contrast to many other viruses that escape from cytotoxic T cell recognition by down-regulating major histocompatibility complex class I-restricted antigen presentation, flavivirus infection of mammalian cells up-regulates cell surface expression of major histocompatibility complex class I molecules. Two putative mechanisms for flavivirus-induced major histocompatibility complex class I up regulation, one via activation of the transcription factor NF-kappaB, the second by augmentation of peptide import into the lumen of the endoplasmic reticulum, are reviewed, and the biological effect of the flavivirus-mediated phenomenon on target cell recognition by natural killer and cytotoxic T cells is addressed. Finally, we speculate on the physiological role of flavivirus-mediated modulation of major histocompatibility complex class I antigen presentation in the context of the biology of flavivirus transmission between the vertebrate host and arthropod vector and suggest that it may represent a strategy for immune evasion from the natural killer cell response or, alternatively, that up-regulation of major histocompatibility complex class I is a by-product of flavivirus replication without significance for virus growth. PMID- 12752687 TI - Is flavivirus resistance interferon type I-independent? AB - Interferon type I comprises a group of major virus-inducible host antiviral factors that control infection with a great number of human and animal viruses. They are ubiquitously expressed cytokines that interfere with virus replication within different cell types by activating a number of host genes and several parallel antiviral pathways. Two major intracellular actors of IFN-I-induced antiviral states are ribonucleic acid-dependent protein kinase and 2'-5' oligoadenylate synthetases/RNase L, both being induced by IFN-I and activated by viral double stranded ribonucleic acid. In addition, Mx proteins and ribonucleic acid-specific adenosine deaminase have also been implicated in IFN-I-induced antiviral responses to some RNA viruses. Viruses, in turn, have evolved different strategies to escape a control imposed by IFN-I and by IFN-I-induced antiviral factors. The fatal outcome of virus infection as well as the efficiency of IFN-I based antiviral therapies in its prevention, are determined by complex interactions between viral virulence factors and cellular antiviral IFN-I inducible factors. In the light of these facts and current knowledge on IFN-I involvement in flavivirus infection, I discuss a possible role of IFN-I signalling in resistance to flavivirus infection in a model of congenic mouse strains that express different levels of susceptibility/resistance to common flaviviruses. Specifically, this review emphasizes importance of fully operative 2'-5'-oligoadenylate synthetases/RNase L pathway for the IFN-I-induced stimulation of flavivirus resistance conferred by Flv. PMID- 12752688 TI - Infection of mouse neurones by West Nile virus is modulated by the interferon inducible 2'-5' oligoadenylate synthetase 1b protein. AB - Over the past 7 years, West Nile zoonosis has been an emerging concern for public health in Europe, Middle East and more recently in North America. West Nile virus causes epidemic outbreaks in humans and infected patients may exhibit severe neurological symptoms. Because susceptibility and sensitivity to West Nile virus infections may depend on host genetic factors, a mouse model has been established to investigate the genetic determinism of host susceptibility to West Nile virus. A nonsense mutation in gene encoding the 1b isoform of the 2'-5'oligoadenylate synthetase (OAS1b) was constantly associated with the susceptibility of mouse strains to experimental West Nile virus infection. Oligoadenylate synthetase are interferon-inducible proteins playing a role in the endogeneous antiviral pathway. It was of interest to establish whether interferon-alpha and OAS 1B were sufficient to mediate resistance to West Nile virus infection. In the present study, we showed that interferon-alpha had the ability to modulate West Nile virus infection in mouse. In vitro, interferon-alpha protected mouse neuroblastoma cells against West Nile virus infection if cells have been pretreated with the cytokine for several hours. As a consequence of the presence of a stop codon, the Oas1b gene of the susceptible mice encodes a truncated and presumably inactive form, while resistant mice have a normal copy of the gene. Stable mouse neuroblastoma cell clones overexpressing mutant or wild-type OAS 1B were established. Replication of West Nile virus was less efficient in cells that produce the normal copy of OAS 1B as compared to those expressing the truncated form. Our data illustrate the notion that interferon-alpha and Oas genes may be critical for West Nile virus pathogenesis. PMID- 12752726 TI - Helicobacter pylori infection and perforated peptic ulcer prevalence of the infection and role of antimicrobial treatment. AB - Although the role of Helicobacter pylori infection on noncomplicated peptic ulcer disease has been definitively established, the precise relationship between the organism and complicated ulcer has hardly been studied. The mean prevalence of H. pylori infection in patients with perforated peptic ulcer is of only about 65 70%, which contrasts with the almost 90-100% figure reported in noncomplicated ulcer disease. However, H. pylori infection rates in various studies range markedly from 0% to 100%, suggesting that differences in variables as number and type of diagnostic methods used to diagnose H. pylori infection, or frequency of nonsteroidal anti-inflammatory drug intake, may be responsible for the low prevalence reported in some studies. Recurrent ulcer disease after peptic ulcer perforation mainly occurs in patients with H. pylori infection, which suggests that the microorganism plays an important role in this complication. All patients with perforated peptic ulcer should be treated by simple closure of the perforation and with therapy aimed at healing of the ulcer and eradicating the H. pylori infection, as disappearance of the organism prevents, or at least decreases, ulcer recurrence and ulcer perforation in patients with H. pylori associated perforated ulcers after simple closure. Therefore, H. pylori eradicating treatment should be started during the immediate postoperative period. The patients with intractable recurrent symptoms of peptic ulcer despite adequate medical treatment, but without H. pylori infection (e.g. a patient using nonsteroidal anti-inflammatory drugs), is probably the only remaining indication for elective definitive surgical treatment of peptic ulcer disease. PMID- 12752727 TI - Helicobacter pylori and hepatitis A virus infection in school-aged children on two isolated neighborhood islands in Taiwan. AB - BACKGROUND: The transmission routes of Helicobacter pylori and hepatitis A virus (HAV) infections have been extensively discussed in previous literature. However, whether H. pylori and HAV shared the same transmission pattern or not remains unclear. Lower socioeconomic status was recognized as a consistent risk factor to both infections. However, whether fecal-oral transmission was a risk factor to both infections is still under debate. MATERIALS AND METHODS: In 1996, we conducted a cross-sectional study to evaluate the seroprevalence of antibody to H. pylori and HAV among the randomly selected school-aged children (age between 13 and 15) on Green Island (n = 91) and Lanyu Island (n = 138) (two isolated neighborhood islands near Taiwan Main Island). RESULTS: The seroprevalence of H. pylori and HAV on the Green Island were 82.4% and 5.5%, respectively. The seroprevalence of H. pylori and HAV on Lanyu Island were 71.0% and 90.6%, respectively. H. pylori seroprevalence of all children and the subgroup of 13 year-olds was significantly lower on Lanyu Island than Green Island. However, it was not significantly different in subgroups of 14- and 15-year-olds. HAV seroprevalence was significantly higher on Lanyu Island than Green Island among all children and in each age subgroup. The correlation of H. pylori infection and HAV infection did not demonstrate significant linear correlation on both islands. CONCLUSIONS: In conclusion, H. pylori and HAV infections in school-aged children of 13-15 years of age on Green Island and Lanyu Island did not demonstrate significant correlation. The results of this study imply that H. pylori and HAV may share different transmission routes of infection. PMID- 12752728 TI - Serologic prevalence of antibodies to Helicobacter pylori in internationally adopted children. AB - BACKGROUND: Helicobacter pylori (H. pylori) infection has been linked to gastritis, diarrhea, peptic ulcers, failure-to-thrive, anemia, as well as predisposition to gastric malignancies. Because many internationally adopted children have diarrhea, failure-to-thrive, and anemia on arrival to the US, we determined the prevalence of HP antibodies among these children. METHODS: Serum samples from 226 unselected children from 18 countries who were evaluated in the International Adoption Clinic at New England Medical Center were tested for antibodies to H. pylori. The results of serologic screening were analyzed in relation to age at adoption, site of residence prior to adoption, weight and height, and the presence or absence of anemia, diarrhea, or intestinal parasites. RESULTS: 31% of internationally adopted children had antibodies to H. pylori. The presence of H. pylori-antibodies was associated with residence in an orphanage (vs. foster care) prior to adoption, older age at adoption, and coinfection with intestinal parasites. No direct effects on height or weight were identified; no associations with diarrhea or anemia were found. CONCLUSIONS: Internationally adopted children have a high incidence of exposure to H. pylori, as diagnosed serologically. Residence in an orphanage (compared with foster care), older age at adoption, and coinfection with intestinal parasites were more common among children seropositive for anti-H. pylori antibodies. PMID- 12752729 TI - The association of intestinal parasitosis and H. pylori infection in children and adults from a Mexican community with high prevalence of parasitosis. AB - BACKGROUND: Experimental evidences have suggested that a Th1 response is unable to eliminate H. pylori colonization; whereas a Th2 response, like the one induced by vaccination, reduces H. pylori infection in animal models. Some parasitic infections induce a polarized Th2 response, which theoretically would favor a reduced H. pylori prevalence. The aim of this work was to study the possible association between parasitic infections and H. pylori prevalence. MATERIALS AND METHODS: The study population included 120 children and 188 adults from a low socioeconomic level village. H. pylori prevalence was determined in serum by ELISA; parasitic infections were identified in feces by microscopic examination; and total serum IgE levels, as an indirect indicator of some parasitic infections, were determined by ELISA. RESULTS: In children, H. pylori prevalence was no different between those with and without intestinal parasitic infection. By contrast, adults with intestinal parasitic infection had a significantly lower H. pylori prevalence than adults without parasites (62.6% compared with 80.4%; p = 0.006, OR 2.45). Also in adults, but not in children, total IgE levels were significantly higher in those without H. pylori infection than in those with H. pylori infection (p < 0.001). CONCLUSIONS: Intestinal parasitic infections and serum IgE levels showed an age-dependent association with H. pylori prevalence. In adults, but not in children, intestinal parasitic infections and increased IgE levels where associated with a reduced H. pylori prevalence. PMID- 12752730 TI - Is the recurrence of Helicobacter pylori infection after eradication therapy resultant from recrudescence or reinfection, in Japan. AB - BACKGROUND: Reinfection of Helicobacter pylori after eradication is rare in developed countries but most often occurs within 1 year. In the present study, we attempted to differentiate between reinfection and recrudescence of H. pylori strains between 6 months and 6 years after successful eradication in Japan, a country with a high prevalence of H. pylori infection. MATERIALS AND METHODS: After successful eradication of H. pylori, 274 patients were followed up by endoscopy and urea breath test. In recurrent patients, H. pylori strains isolated initially and after recurrence were compared using PCR-based restriction fragment length polymorphism (RFLP) analysis. RESULTS: Recurrence of H. pylori occurred in 15 of 274 patients (5.5%) at 6 months after eradication and the annual recurrence rate was 2.0% per patient year (between 1 and 6 years). PCR-based RFLP analysis of H. pylori strains isolated initially and after recurrence showed that 62.5% (at 6 months) and 100% (after 1 years) of bacteria were of different strains. CONCLUSION: Reinfection of H. pylori was not as rare at 6 months after eradication as reported previously, and up to 6 years after eradication, the annual reinfection rate is 2.0% per patient year in Japan. PMID- 12752731 TI - Prevention and suppression of Helicobacter felis infection in mice using colostral preparation with specific antibodies. AB - BACKGROUND: Specific antibodies against Helicobacter were enriched from the colostra of hyperimmunized cows. Efficacies of colostral control preparation and immune preparation containing specific antibodies against Helicobacter felis were studied in the prevention and treatment of experimental H. felis infection in mice. MATERIALS AND METHODS: H. felis-infected mice were given either immune or control preparation with or without complement or amoxicillin orally in four different trials. H. felis status was assessed on the basis of bacterial stainings, gastric histology and serum antibodies. RESULTS: Immune, but not control preparation, prevented H. felis infection (p > 0.01), the efficacy being dependent on the presence of specific antibodies. In the trial on infected Balb/c mice treatment with immune preparation (p = 0.029) but not control preparation decreased the colonization of gastric antrum by H. felis. In the further trials with infected SJL-mice, treatments with colostral preparations did not decrease colonization. Amoxicillin treatment decreased the colonization with trend-setting significance (p = 0.056; infected mice as controls), whereas amoxicillin combined with immune preparation had a significant effect (p < 0.0005). CONCLUSIONS: Specific colostral antibodies were useful in the prevention of Helicobacter infection in a mouse model. The results of the treatment trials were controversial but a similar colostral immune preparation against H. pylori could be effective and useful in preventing infections in humans and during antibiotic treatment. PMID- 12752732 TI - Mixed-infection of antibiotic susceptible and resistant Helicobacter pylori isolates in a single patient and underestimation of antimicrobial susceptibility testing. AB - Antibiotic resistance among Helicobacter pylori has been increasing worldwide and has begun to affect the overall efficacy of current antibiotic regimens adversely. We examined 220 pairs of H. pylori isolates obtained from both the antrum and corpus of separate patients; 109 (50%) harbored antibiotic-resistant H. pylori: amoxicillin (0.5%), clarithromycin (5.9%), furazolidone (1.4%), metronidazole (45.5%), nitrofurantoin (1.4%), and tetracycline (6.8%). Heteroresistance among the two biopsy sites from each patient was present in 41 of the 109 patients (38%) with antibiotic resistant H. pylori (e.g. 34% with resistant strains would be misclassified as susceptible if a biopsy of the antrum alone used for antimicrobial susceptibility testing). DNA fingerprinting genotype analysis was carried out on the 41 pairs of isolates with heteroresistance. While different patients had different fingerprinting patterns, each pair of isolates showed identical or similar fingerprinting patterns. These results suggest that antibiotic-resistant H. pylori typically develop from pre-existing susceptible strain rather than coinfection with a different strain. The minor differences in genotype (degeneration of genotype) seen reflect one of the processes for development of genetic diversity in H. pylori. No biopsy single site can be considered representative for antimicrobial susceptibility testing. PMID- 12752733 TI - Antimicrobial activity of essential oils against Helicobacter pylori. AB - BACKGROUND: Helicobacter pylori is an important pathogen responsible for gastroduodenal diseases in humans. Although the eradication of H. pylori using antibiotics often improves gastroduodenal diseases, resistance to the antibiotics is emerging. MATERIALS AND METHODS: The antimicrobial effect of essential oils and the development of resistance to the essential oils were evaluated in vitro and in vivo. RESULTS: Thirteen essential oils used in this study completely inhibited the growth of H. pylori in vitro at a concentration of 0.1% (v/v). Cymbopogon citratus (lemongrass) and Lippia citriodora (lemon verbena) were bactericidal against H. pylori at 0.01% at pH 4.0 and 5.0. Resistance to lemongrass did not develop even after 10 sequential passages, whereas resistance to clarithromycin developed under the same conditions. In in vivo studies, the density of H. pylori in the stomach of mice treated with lemongrass was significantly reduced compared with untreated mice. CONCLUSIONS: These results demonstrate that the essential oils are bactericidal against H. pylori without the development of acquired resistance, suggesting that essential oils may have potential as new and safe agents for inclusion in anti-H. pylori regimens. PMID- 12752734 TI - Helicobacter pylori modulation of gastric and duodenal mucosal T cell cytokine secretions in children compared with adults. AB - BACKGROUND: In contrast to adults, ulcers are un-common in Helicobacter pylori infected children. Since immunological determinants influence the outcome of H. pylori infection, we have investigated mucosal T cell responses in H. pylori infected children and compared them with those of adults and negative controls. MATERIAL AND METHODS: Mucosal biopsies were obtained from 43 patients undergoing an upper GI endoscopy for dyspeptic symptoms. The concentrations of released cytokines and the density of CD3+, CD25+ and CD69+cells were evaluated by flow cytometry, and the numbers of cytokine-secreting cells were measured by ELISPOT. RESULTS: The numbers of isolated antral CD3+ lymphocytes were only significantly raised in infected adults compared with noninfected controls (p < 0.05), whereas the proportion of CD3+ cells expressing activation markers (CD25 or CD69) remained low. In the stomach, IFN-gamma concentrations increased in infected children and infected adults compared with controls (p < 0.05), but IFN-gamma concentrations were tenfold lower in children than in adults (p < 0.01). IL-2, IL 4, IL-10 and TNF-alpha concentrations were similar in infected and in uninfected children and adults. In contrast, in the duodenum, IFN-gamma, as well as IL-4 and IL-10 concentrations were only increased in infected children compared with controls (p < 0.05). The concentrations of these cytokines were similar in both groups of adults who, however, like children, displayed a higher number of duodenal IL-4-secreting cells compared to controls (p < 0.05). CONCLUSION: These results suggest that IFN-gamma secretion in the stomach of H. pylori-infected patients is lower in children than in adults. This could protect children from development of severe gastro-duodenal diseases such as ulcer disease. In addition, infected patients are characterised by a dysregulation of the mucosal cytokine secretion at distance from the infection site. PMID- 12752735 TI - Helicobacter pylori CagA status, mucosal oxidative damage and gastritis phenotype: a potential pathway to cancer? AB - BACKGROUND: Oxidative DNA damage is associated with Helicobacter pylori infection, atrophy and intestinal metaplasia. H. pylori-cagA-positive strains are associated with the highest risk of gastric cancer. AIMS: To ascertain whether cagA-positive H. pylori infection correlates with higher concentrations of 8OHdG and the presence of precancerous changes. PATIENTS AND METHODS: 118 patients were studied (65M/53F, age 61 +/- 14 years). Twelve were H. pylori-negative. Among the H. pylori-positive patients, 34 were cagA-positive and 40 were cagA negative. In 32 patients H. pylori had been eradicated at least 6 months before endoscopic sampling. The phenotype of the gastritis (atrophic compared with nonatrophic, with and without intestinal metaplasia) was scored in biopsy samples obtained from the antrum, corpus, and angularis incisura. In antral biopsy samples, 8 hydroxydeoxyguanosine was assessed by HPLC (electrochemical detector). CagA status was determined by PCR. RESULTS: The highest scores for both mononuclear inflammation and activity of gastritis were significantly associated with cagA status (p = 0.036 antrum and p = 0.02 corpus). cagA-positive infection significantly correlated with a higher prevalence of atrophic-metaplastic lesions (p = 0.04). cagA-positive patients had higher 8-hydroxydeoxyguanosine levels than both cagA-negative and H. pylori-negative cases (p = 0.01). The 8 hydroxydeoxyguanosine levels were significantly higher in multifocal atrophic gastritis (p = 0.04). The odds ratio for cagA-positive patients having 8OHdG levels above a cut-off calculated on the basis of the ROC curves were 7.12, overall, reaching 11.25 when only patients younger than 50 were considered. CONCLUSIONS: cagA-positive patients were characterized: first, for higher scores for gastritis, activity and atrophic and metaplastic lesions; and second for greater oxidative DNA damage overall, at younger age and in the presence of multifocal atrophy. This setting may represent a cancer-prone biological context. PMID- 12752736 TI - CagA tyrosine phosphorylation in gastric epithelial cells caused by Helicobacter pylori in patients with gastric adenocarcinoma. AB - BACKGROUND: Tyrosine phosphorylation of Helicobacter pylori cytotoxin-associated protein of in gastric epithelial cells is reported. The goals of this study are first to examine the occurrence of CagA tyrosine phosphorylation in H. pylori strains isolated from patients with gastric adenocarcinoma and gastritis, and second to clarify the relationship between the diversity of tyrosine phosphorylation motifs and the presence of CagA tyrosine phosphorylation. METHODS: Fifty-eight clinical isolates of H. pylori from patients with gastric adenocarcinoma (29 cases) and gastritis (29 cases) were studied for CagA tyrosine phosphorylation by Western blotting. Sequence diversity of tyrosine phosphorylation motifs was analysed among positive- or negative-CagA tyrosine phosphorylation isolates. RESULTS: Positive CagA tyrosine phosphorylation was found in 93.1% (27 of 29) of strains from gastric adenocarcinoma patients and 51.7% (15 of 29) of strains from gastritis patients (p < 0.001). Intact motifs were found in H. pylori isolates with CagA tyrosine phosphorylation. Of the 16 negative CagA tyrosine phosphorylation isolates, intact tyrosine phosphorylation motifs were found in 15 isolates. CONCLUSIONS: CagA tyrosine phosphorylation, which is significantly greater in strains from gastric adenocarcinoma patients, may play a role in gastric carcinogenesis, and could be a better marker of more virulent strains than the cag pathogenicity island in Asia, where the cag pathogenicity island is present in nearly all H. pylori strains. Sequence diversity of tyrosine phosphorylation motifs on CagA was not related to the presence of tyrosine phosphorylation. The absence of tyrosine phosphorylation motif might result in negative tyrosine phosphorylation phenotypes, but such motifs are not the sole factors associated with CagA tyrosine phosphorylation. PMID- 12752737 TI - Patient's information environments: deserts, jungles and less hostile alternatives. PMID- 12752738 TI - A survey of the decision-making needs of Canadians faced with complex health decisions. AB - OBJECTIVE: To describe the decision-making needs of Canadians when faced with 'complex' health decisions characterized by balancing advantages against disadvantages. Although a national report emphasized that public confidence in the health-care system depends on support for personal knowledge and decision making, there has been no systematic investigation of the Canadian population's decision-making needs. DESIGN: Cross-sectional telephone survey using random digit dialling. PARTICIPANTS: National sample of 635 adults over 18 years of age, living in Canada. RESULTS: Forty-two percentage of eligible contacts participated. Sixty-five percent of contacts reported making 'complex' health decisions, commonly about medical or surgical treatments or birth control, and more commonly by women and by married/separated individuals. Most respondents took an active role in their decisions, often sharing the process with their partner or family. Being younger was associated with a more independent role. Physicians were more often involved in the decisions of respondents with less education. Fifty-nine percent of respondents experienced decisional conflict; more conflict was seen with those who were female and feeling uninformed about options, pressured to select one particular option, and unready or unskilled in decision-making. Less decisional conflict was seen in those who reported birth control decisions and in those who were 70 years and older. Participants used several strategies when deliberating about choices including: information gathering, clarifying their values, and seeking support and information from others. Personal counselling and printed information materials were commonly preferred methods of learning about options. 'Essential' criteria for judging satisfactory decision-making included: having sufficient knowledge about the options, outcomes, and probabilities; being clear about values; selecting and implementing a choice that agrees with personal values; and expressing satisfaction with the choice. CONCLUSIONS: Canadians, particularly women, face difficult decisions and need support and information from credible sources. PMID- 12752739 TI - What factors are associated with a woman's decision to take hormone replacement therapy? Evaluated in the context of a decision aid. AB - OBJECTIVES: To understand the factors associated with a post-menopausal woman deciding to take hormone replacement therapy (HRT) after reviewing a decision aid (DA) and having a counselling visit with her physician as well as the factors associated with the act of taking HRT 2 months after the counselling interview. DESIGN: A secondary analysis of data collected for a randomized controlled trial evaluating two DAs. MAIN OUTCOME RESULTS: Although 28% of women were uncertain regarding their decision after the counselling interview, only 2.4% of women, at the assessment at 2 months, had not made a decision. The most significant factor associated with the decision to take HRT, after the physician visit, was the physician preference (OR: 62, 95% CI: 13.3, 289.7). Physician preference (OR: 78, 95% CI: 6.2, 975) remained the most significant factor for taking HRT 2 months after the counselling interview followed by low uncertainty about the decision (OR: 0.4, 95% CI: 0.2, 0.7). CONCLUSION: Physician preference was the factor that was most associated with the woman's decision following counselling and 2 months later. Qualitative evaluation of the interview process involving the patient and physician would determine whether the patient and physician are reaching a shared decision or is the physician preference influencing the patient. PMID- 12752740 TI - Patient participation in decision-making on the introduction of home respiratory care: who does not participate? AB - OBJECTIVES: In this study we examined home respiratory patients' participation in decision-making on whether to begin home respiratory care therapy, and this participation in decision-making during the latest visit to the clinic. SUBJECTS AND METHODS: The target population consisted of patients who were using home respiratory care devices and who were visiting the outpatient clinics. Postal questionnaires were sent to 4159 patients (40% of respiratory care device users in Finland). A total of 3336 answered (response rate 80%) and 3153 were eligible for analysis. Odds ratios, chi-square tests, Mann-Whitney U-test and stepwise logistic regression analysis were used in the data analyses. RESULTS: Patients who did not participate in decision-making were more frequently older people, women and had lower income than the other patients. While these results parallel those of previous studies, in contrast we found more women with high education to be non-participants. Non-participants were not participating in decision-making during their latest visit to the clinic in spite of the fact that they considered participation almost as important as did the other patients. Non-participants were less satisfied with the quality of care given and felt that their life had improved less than did the other patients. CONCLUSION: The ethical principle of equal opportunities to participate in care decisions was not applied among home respiratory care patients in this study. The results challenge health-care professionals to notice inequalities and improve their practices. The results can be generalized to all home respiratory care patients in Finland. PMID- 12752741 TI - Elaborating patient information with patients themselves: lessons from a cancer treatment focus group. AB - OBJECTIVE: To assess the significance of patients' input in the elaboration of a patient information booklet. DESIGN: Qualitative study based on focus group discussions. SETTING: Centre Leon Berard, a comprehensive cancer centre in the Rhone-Alpes region of France. PARTICIPANTS: (1) A multidisciplinary working group (oncologists, health economists and one clinical psychologist) wrote up initial information documents concerning possible breast cancer treatments. (2) A focus group comprised of patients with a history of breast cancer and healthy volunteers discussed their reactions to these documents. MAIN OUTCOME MEASURE: Analysis of the focus group's reactions according to key themes predetermined by the working group and related themes introduced by the focus group itself. RESULTS: The focus group proposed numerous, significant modifications to answer requests for additional information, clarification and better readability in the information booklets. DISCUSSION/CONCLUSIONS: This qualitative analysis showed a significant input of patients' perspective in the elaboration of patient information. It is also an additional support to the feasibility and appropriateness of the focus group technique. The next stage will be to test whether information documents produced here conform to the needs of patients currently undergoing treatment. PMID- 12752742 TI - The use of willingness to pay to assess public preferences towards the fortification of foodstuffs with folic acid. AB - OBJECTIVES: To assess public attitudes towards the fortification of flour with folic acid, and quantify their intensity of preference towards the proposed policy. DESIGN: Structured interviews describing the proposed policy of fortification followed by questions about the respondent's preferences towards fortification. SETTING AND PARTICIPANTS: A United Kingdom community sample of 76 people interviewed at home. MAIN VARIABLES STUDIED: Direction of preference towards fortification, willingness to pay (WTP) for the preferred course of action, and the reasons behind their preferences. RESULTS: Responses showed that 51 (67%) were in favour of fortification, 15 (20%) were opposed, while the remaining 10 (13%) were either indifferent or were unsure. Those in favour of fortification tended to be younger and poorer than those opposed to it. Willingness to pay estimates show that those in favour of food fortification had more intense preferences, with mean and median WTP around twice as great. Reasons for being willing to pay were centred on the health benefits, with particular reference being made to the intervention saving lives and it being preventative. Those opposed tended to believe that there was insufficient evidence. CONCLUSIONS: The combination of a policy vote, WTP and qualitative data, allow us to assess the direction, intensity and motivations behind people's preferences. Further work needs to be undertaken to gather more robust estimates of public preferences for fortification, and to better understand attitudes towards public health interventions more generally. PMID- 12752743 TI - Preliminary validation of the Satisfaction With Decision scale with depressed primary care patients. AB - OBJECTIVE: To conduct a preliminary validation of the Satisfaction With Decision (SWD) scale with depressed primary care patients. DESIGN: Cross-sectional observational pilot study using a postal survey. SETTING AND PARTICIPANTS: Depressed primary care patients (n = 97) who recently had made a new decision about antidepressant medication use completed surveys regarding their treatment decisions. MAIN VARIABLES: Measures included patient-reported satisfaction with decision, decisional conflict, knowledge about depression and treatment, decision involvement, pain and health status, antidepressant medication efficacy, and satisfaction with health services. RESULTS: The SWD scale had good internal consistency reliability (alpha = 0.85). Evidence for construct validity was confirmed via a hypothesized pattern of relationships between the SWD scale and other measures. Decision satisfaction was associated with several issues of relevance for designing patient-centred decision support interventions: (1) knowledge about depression and treatment; (2) involvement in health-related decisions; and (3) aiding evaluation of trade-offs among pros and cons of treatment. CONCLUSIONS: The results of this pilot study show that the SWD scale appears to be a psychometrically sound and practical measure for research with this population. Additional research is needed on the theoretical nature of decision satisfaction and developing and testing patient-centred decision support interventions for depression treatment. PMID- 12752745 TI - Monitoring health information services: a new tool for information quality review. PMID- 12752744 TI - Patient characteristics as predictors of primary health care preferences: a systematic literature analysis. AB - OBJECTIVE: To identify associations between various cultural and demographic factors and patients' primary health care preferences. SEARCH STRATEGY: Searches were performed in MEDLINE (1966-December 2000), PsycINFO (1977-May 2001) and Sociological Abstracts (1963-December 2000). Identified papers were checked for more papers. INCLUSION CRITERIA: Studies with a focus on primary health care or health care in general, asking patients about preferences with regard to health care, reporting quantitative results and examining the relations between specific patient characteristics and patient preferences. DATA EXTRACTION AND SYNTHESIS: Data were extracted from studies using a scoring form to register what methods were used, which patient characteristics were analysed and which patient characteristics significantly influenced patients' preferences with regard to different aspects of health care (P < 0.05). MAIN RESULTS: A total of 145 studies were included with 2276 comparisons between subgroups of patients. Of all the comparisons, 607 (27%) showed a significant association between patient characteristics and preferences with regard to primary health care. Age and economic status significantly related to patient preferences in 38 and 33% of the comparisons, respectively. Education, health status, family situation, sex, and utilization of health care related significantly to patient preferences in less than 25% of the comparisons. CONCLUSIONS: This review of the literature showed patient characteristics to be an important determinant of preferences regarding many aspects of primary health care defined as general practice care or health care, in general. All of the patient characteristics examined here showed at least some significant associations with preferences for primary health care. PMID- 12752747 TI - Center of excellence for headache care: group model at Kaiser Permanente. AB - OBJECTIVE: To evaluate the effectiveness of a disease management model for primary headache by: (1) assessing improvement in patients' quality of life, (2) decreasing headache-related visits to primary care and emergency departments, and (3) maintaining high levels of patient and physician satisfaction. BACKGROUND: Patients with headache regularly seek health care but, in general, are dissatisfied with the care they receive. Patients with primary headaches utilize resources and cost health plans more than patients with other chronic diseases. Primary care visits are time restricted, prohibiting adequate headache evaluation and management. Practice guidelines are inconsistently followed, and access to headache specialists is limited. This headache management program implemented an alternative means of delivering care to manage large volumes of patients with headache. A multidisciplinary team approach coordinated by a neurologist, utilizing education and a nurse practitioner as the main provider of care, was the central process of the program. METHODS: This was a pilot study involving a prospective cohort with defined outcome measures. Inclusion criteria were adult patients with primary headaches. Patients initially attended an educational session instructed by a neurologist and a nurse practitioner. The patient was subsequently evaluated by the nurse practitioner who developed and coordinated a comprehensive individual treatment plan. The Migraine-Specific Quality of Life and the Medical Outcomes Study 36-Item Short Form Questionnaires were completed at baseline, at follow-up visits, and 6 months after completion of the program. Subjective patient assessment of improvement in their headaches, chart review for tabulation of headache-related visits, and primary care physician satisfaction surveys were measured. RESULTS: Both the Migraine-Specific questionnaire and the Short Form-36 measurements demonstrated a statistically significant improvement at 8 weeks, and this was maintained for 6 months after completing the program. At completion of the program, 92% of patients reported subjective improvement. Patient visits for headaches to primary care and emergency departments showed a significant decrease. High levels of satisfaction for primary care physicians were achieved. CONCLUSIONS: A disease management model using a multidisciplinary team improved individualized patient care. This model increased patient/provider rapport and communication through an educational class. It empowered the patient to take control of their health care by utilizing shared decision making. Patient satisfaction improved and overall health care utilization was reduced. PMID- 12752748 TI - Rapid and sensitive paradigm for screening patients with headache in primary care settings. AB - OBJECTIVE: To determine the sensitivity and specificity of a brief headache screening paradigm for primary care clinicians. BACKGROUND: Migraine and drug rebound headache are disabling primary headache disorders. Both are underdiagnosed and undertreated. A method for rapid screening of migraine, drug rebound headache, and other daily headache syndromes would be useful. The Brief Headache Screen uses 3 questions-the frequency of severe (disabling) headache, other (mild) headache, and use of symptomatic medication-to generate diagnoses. METHODS: The Brief Headache Screen was evaluated in an emergency department, a family practice department, and a referral headache clinic. Diagnoses from the Brief Headache Screen were compared to diagnoses of trained researchers and headache specialists. RESULTS: Three hundred ninety-nine patients were screened and interviewed. The criterion of episodic severe (disabling) headache correctly identified migraine in 136 (93%) of 146 patients with episodic migraine and 154 (78%) of 197 patients with chronic migraine, with a specificity for any migraine (episodic or chronic) of 32 (63%) of 51. The inclusion of episodic or daily severe headache identified migraine in 100% of patients with chronic migraine. Only 6 (1.7%) of 343 patients with migraine were not identified by severe (disabling) headache. The combination of severe and mild headache frequency was sensitive to daily headache syndromes in 218 (94%) of 232 patients with a specificity of 87 (54%) of 162. Medication overuse was correctly identified in 146 (86%) of 169 patients with a specificity of 22 (79%) of 28. CONCLUSIONS: The frequency of severe (disabling) and mild headaches and use of symptomatic medications, rapidly and sensitively screens for migraine, daily headache syndromes, and medication overuse. The use of this paradigm in primary care settings may improve the recognition of these important headache syndromes. PMID- 12752749 TI - Demographic and migraine characteristics of adolescents with migraine: Glaxo Wellcome clinical trials' database. AB - OBJECTIVE: To describe the demographics and migraine characteristics of patients in the Glaxo Wellcome adolescent clinical trials' database. METHODS: Data from 8 sumatriptan (tablet and nasal spray) and naratriptan (tablet) trials (6 placebo controlled and 2 open label) were reviewed. Adolescents aged 12 to 17 years who had participated in migraine clinical trials and used at least 1 dose of study medication were summarized using descriptive statistics. Patient demographic (gender, age, race, height, and weight) and migraine (diagnosis, pain location and intensity, time and day of migraine onset and treatment, and associated symptoms) characteristics were examined. RESULTS: One thousand nine hundred thirty-two adolescents with migraine were identified; mean age was 14.1 years (standard deviation, 1.64; range, 11 to 18) and 54% of patients were female. More males were represented in the 12- to 14-year-old group (646 [73%] of 885) than in the 15- to 17-year-old group (234 [26%] of 885). Most patients reported migraine without aura (67%, 1121 of 1672), unilateral migraine pain (58%, 458 of 787), and pulsating pain (74%, 582 of 790). Migraine was aggravated by physical activity in most of the adolescents (88%, 526 of 598). Most migraine attacks (73%, 1363 of 1858) began between 6 am and 6 pm, and proportionately more attacks occurred Monday through Wednesday. Pretreatment vomiting was experienced by 5% (97 of 1830) of patients, nausea by 53% (983 of 1849), and photophobia or phonophobia (or both) by 88% (1628 of 1858) of patients. The incidence of associated symptoms was directly related to pretreatment headache severity. CONCLUSIONS: In this large clinical trials' database, adolescents had migraine without aura characterized by unilateral and pulsating pain and aggravated by activity. The incidence of associated symptoms was directly related to pretreatment pain intensity. More migraines occurred Monday through Wednesday during typical school hours. These data may facilitate clinicians' efforts to tailor migraine therapy to the needs of this patient population. PMID- 12752750 TI - Cerebrovascular reactivity in adolescents with migraine and tension-type headache during headache-free interval and attack. AB - BACKGROUND: Migraine is a common cause of headache in adolescents. Assuming that the cerebral vasculature is involved in the pathophysiology of migraine, we compared cerebral vasoreactivity in adolescents both during a migraine attack and a headache-free interval. METHODS: A functional transcranial Doppler test utilizing a visual stimulation paradigm was undertaken to measure the evoked flow velocity in the posterior cerebral artery of adolescents suffering from a migraine without aura or a tension-type headache. To serve as a control, data previously obtained from age-matched adolescents with no primary headache disorder were used. The flow curves were evaluated by determining the maximal flow velocity increase and by modeling their time course according to a control system analysis. In that analysis, the main dynamic features of the flow response were described mathematically in terms of a control system model of low order. The parameters were time delay, gain, attenuation, rate time, and natural frequency. RESULTS: The attenuation parameter (P<.005), indicative of an increased tone of the vessel, and the resting absolute flow velocity (P<.01) both showed a significant increase during an attack in the adolescents with migraine; the gain parameter showed a trend towards similar increase (P =.07). The maximal flow velocity did not increase significantly during an attack. CONCLUSIONS: The control system approach utilized here appears to be more sensitive for detecting migraine-associated changes in cerebral vasoreactivity than examination of the maximal flow velocities alone. PMID- 12752751 TI - Normalization of high interictal cerebrovascular reactivity in migraine without aura by treatment with flunarizine. AB - BACKGROUND: Modification of migraine-associated cerebrovascular reactivity may provide insight into the mechanism of action of a given therapeutic intervention. METHODS: With transcranial Doppler and a breath-holding index, cerebrovascular reactivity to hypercapnia was evaluated in 20 patients with migraine without aura interictally and in 11 healthy controls. Patients were started on prophylactic treatment with flunarizine 10 mg per day, and measurements were repeated at the end of every month for 3 months. Headache status was evaluated clinically via a headache index. Headache index; breath-holding index; systolic, diastolic, and mean blood flow velocities; and pulsatility index measurements were recorded at every session. RESULTS: The baseline breath-holding index was significantly higher in the migraine group compared to the control group (P =.002). No difference in other parameters was found between the groups. The change in the headache index was significant (P<.001), indicating a beneficial effect from flunarizine. The breath-holding index improved significantly after treatment (P<.001), and the baseline difference in the breath-holding index between the pretreatment migraine group and the control group was no longer evident at 3 months. There was no significant change with treatment in the other transcranial Doppler parameters. CONCLUSIONS: Our finding of unchanged blood flow velocities but normalized cerebrovascular reactivity after treatment suggests that the mechanism of action of flunarizine in migraine does not involve a vasodilatory effect on cerebral vessels. It may be instead that flunarizine modifies cerebrovascular reactivity through its action on centrally located structures that subserve autonomic vascular control. PMID- 12752752 TI - Migraine prevalence during menopausal transition. AB - BACKGROUND: The effect of menopausal transition on the frequency of migraine has never been the focus of a community-based study. METHODS: A cross-sectional community-based survey was undertaken among Chinese women aged 40 to 54 years in Kinmen, Taiwan. Neurologists diagnosed migraine based on the 1988 International Headache Society classification criteria. Menstrual history including a past or current history of premenstrual syndrome was obtained. Serum levels of estradiol and follicle-stimulating hormone were measured. RESULTS: The 1-year prevalence of migraine was 16.5% in the 1436 participants. Among the women who had not had hysterectomies and did not report symptoms of premenstrual syndrome, migraine prevalence did not vary according to menopausal status. In contrast, in women with self-reported premenstrual syndrome, menopausal status was a factor in migraine prevalence: the late perimenopausal group had the highest prevalence (31%) and the spontaneous menopausal group had the lowest (7%). Among all menopausal groups, women who had had hysterectomies reported the highest migraine prevalence (27%), with the highest occurring in those with premenstrual syndrome (44%). The presence of low estrogen (<50 pg/mL) and high follicle-stimulating hormone levels (>30 mIU/mL) was associated with lower migraine prevalence, even in the premenopausal and early perimenopausal women. CONCLUSIONS: Our data supported the clinical impression that migraine prevalence increases before menopause and declines after spontaneous menopause. However, in this study, this trend occurred only in women with increased vulnerability to hormonal change, such as those with premenstrual syndrome. The presence of low estrogen and high follicle-stimulating hormone levels predicted lower migraine prevalence, whereas a history of hysterectomy was related to higher prevalence. PMID- 12752753 TI - Long-term follow-up of chronic cluster headache treated surgically with trigeminal tractotomy. PMID- 12752754 TI - Naratriptan in the preventive treatment of refractory chronic migraine: a review of 27 cases. AB - OBJECTIVE: To review the efficacy of naratriptan as preventive treatment in 27 patients with chronic migraine refractory to other commonly used preventive therapies. BACKGROUND: The treatment of chronic migraine often poses a major challenge to the clinician. Even when given expert care, patients with chronic migraine may continue to have daily or near-daily headaches. METHODS: Clinical records and headache calendars were reviewed of 27 patients fulfilling the following inclusion criteria: (1) aged 18 to 65 years; (2) diagnosis of chronic migraine (formerly transformed migraine), according to the criteria proposed by Silberstein et al; (3) previous failure of at least 4 preventive medications prescribed as part of a management program that included nonpharmacological measures, preventive medication, acute care medication, and detoxification from overused medication; and (4) have used daily naratriptan for no less than 2 consecutive months. The dose of naratriptan prescribed was 2.5 mg twice daily. We considered the following outcomes: (1) frequency of headache, (2) intensity of pain, (3) number of days per month with severe headache, (4) headache index (frequency times intensity), and (5) proportion of patients who reverted to an episodic pattern of pain after 6 months of treatment. RESULTS: There was a statistically significant reduction in the frequency of headache days 2 months (15.3 days versus 24.1 days at baseline, P<.001), 6 months (9.1 days, P<.001), and 1 year (7.3 days, P<.001) after daily treatment with naratriptan was initiated. There was also a statistically significant reduction in the number of days per month of severe pain at 1 month (5.6 days versus 12.5 days at baseline, P<.01), 2 months (5.7 days, P<.01), 6 months (2.8 days, P<.01), and 1 year (2.6 days, P<.01). Similarly, there was a statistically significant reduction in the headache index at 2 months (33 versus 56.4 at baseline, P<.001), 6 months (19.5, P<.001), and 1 year (17.2, P<.001). Of the 20 patients who continued to use naratriptan daily for at least 6 months, 13 (65%) reverted to an episodic pattern of pain (migraine). At 1 year, 11 (55%) still continued to experience episodic headache, 1 (5%) relapsed to chronic migraine, and 2 (10%) were lost to follow up. No patients had intolerability to naratriptan during the treatment period, and no one stopped treatment due to adverse events. CONCLUSIONS: Naratriptan may have a role in the preventive treatment of intractable chronic migraine. Prospective, controlled studies should be considered. PMID- 12752755 TI - Carnitine palmityltransferase II (CPT2) deficiency and migraine headache: two case reports. AB - BACKGROUND: Migraine headache is common and has multiple etiologies. A number of mitochondrial anomalies have been described for migraine, and mitochondrial dysfunction has been implicated as one potential pathophysiological mechanism. Carnitine is used by mitochondria for fatty acid transportation; its deficiency, however, has not been implicated in migraine pathophysiology. METHODS AND RESULTS: Two adolescent girls presented to the Headache Center at Cincinnati Children's Hospital Medical Center with frequent headaches and were diagnosed with migraine by the International Headache Society (IHS) criteria. Both girls had a history of recurrent fatigue, muscle cramps, and multiple side effects from their prophylactic treatment. Carnitine levels were measured and found to be low. Carnitine supplementation was initiated. Both patients had a reduction in headache frequency, as well as an improvement in their associated symptoms and other complaints. A skin and muscle biopsy obtained from one patient revealed a partial carnitine palmityltransferase II deficiency in the muscle only. CONCLUSIONS: Carnitine palmityltransferase II deficiency may represent another etiology for migraine headache, and may be useful in further defining the pathophysiology of migraine. When properly recognized, supplementation with carnitine may improve the outcome of the migraine as well as the carnitine associated symptoms. PMID- 12752756 TI - Transformation into daily migraine with aura following transcutaneous atrial septal defect closure. AB - A link between migraine with aura and cardiac right-to-left shunting has been previously reported. Abortion or decreased frequency of migraine with aura attacks after atrial septal defect closure has been reported in the literature. We report the first case of transformation of migraine with aura into a daily pattern after atrial septal defect closure. A 48-year-old male who had been suffering from rather infrequent attacks of migraine with sensory and visual aura underwent transcutaneous closure of an atrial septal defect. His migraine attacks changed into a daily pattern the day following the procedure and remained so for 6 months. This change in pattern may be related to a changed intra-atrial pressure after the closure or some other unknown factor. PMID- 12752757 TI - Facial pain as the presenting symptom of lung carcinoma with normal chest radiograph. AB - Facial pain is a rare presenting symptom of nonmetastatic lung carcinoma. Referred pain from tumor invasion and compression of the vagus nerve was the presumed cause in the 31 cases published to date. We report 2 additional cases having an unusual clinical feature, namely, both had radiographic evidence of malignancy absent on initial chest films. Severe facial pain in both cases was explained by pulmonary carcinoma detected only through further investigations. From these cases follows the notable conclusion that referred facial pain of malignant origin can occasionally precede the appearance of neoplasm on routine chest films. It is therefore important for physicians to be familiar with the clinical features of this syndrome in order to choose appropriate further diagnostic testing in patients who may be at risk. PMID- 12752758 TI - "Bad Teeth, Bad Headache, Brain Abscess". PMID- 12752759 TI - Confusional migraine or photoepilepsy? PMID- 12752761 TI - Expression of enhancers is altered in Drosophila melanogaster hybrids. AB - The molecular foundations of evolution are difficult to trace because most protein sequences are virtually identical in closely related species. The largest fraction of sequence within the genome, however, is composed of noncoding sequences where regulatory elements locate to various sites. It has been suggested that changes in the activity of these elements may trigger evolutionary change. In Drosophila, the enhancer trap procedure identifies regulatory sequences in the genome after the insertion of a P-element-based construct. We generated new insertions and characterized their expression domains in the adult eye and larval imaginal disks using the white and LacZ reporter genes. Lines with robust expression patterns in D. melanogaster were analyzed in hybrids to test the conservation of regulatory mechanisms between species. Most of the enhancers used in this study modified their expression in hybrids with the mating species D. mauritiana and D. simulans. Expression changes resulted either in gain or loss of expression and were cell-type or hybrid-genome specific. Further characterization of a limited number of enhancers in D. melanogaster showed that expression domains could adapt to changes in cell number during development but not after the completion of cell proliferation. Also, expression of some enhancers appeared to be sensitive to heterochromatin from the Y but not the X chromosome. Taken together, these results demonstrate the high sensitivity of regulatory mechanisms of gene expression as a prime source of evolutionary change and suggest quantitative changes in available transcription factors as one of the mechanisms involved. PMID- 12752762 TI - Heat shock in the developmentally sensitive period of butterfly eyespots fails to increase fluctuating asymmetry. AB - Fluctuating asymmetry (FA) is considered to provide a means of evaluating developmental stability and to reflect an individual's quality or the stress experienced during development. Stress is predicted to increase the phenotypic variation of both FA and trait size. In this study we examined the effect of a particular heat shock on both FA and size of eyespots in the butterfly, Bicyclus anynana. We also examined whether those eyespots thought to be involved in partner choice and sexual selection were particularly sensitive to stress. We applied a heat shock of 39.5 degrees C for 3 h before, during, and after a sensitive period in eyespot development. We examined the FA, variation in FA, size, and variation in size of five eyespots, two on the dorsal forewing (sexually selected traits), two on the ventral forewing, and one on the ventral hindwing (nonsexually selected traits). For each sex and treatment, the heat shock did not result in significant changes in mean trait size and FA nor in the variation of size and FA. There were no differences in the response to the heat shock between sexually and nonsexually selected traits. We discuss how the increased production of heat shock proteins, including HSP60, may have stabilized development and how this might explain the results. PMID- 12752763 TI - Expression of one sponge Iroquois homeobox gene in primmorphs from Suberites domuncula during canal formation. AB - Sponges (Porifera) represent the evolutionary oldest multicellular animals. They are provided with the basic molecules involved in cell-cell and cell-matrix interactions. We report here the isolation and characterization of a complementary DNA from the sponge Suberites domuncula coding for the sponge homeobox gene, SUBDOIRX-a. The deduced polypeptide with a predicted Mr of 44,375 possesses the highly conserved Iroquois-homeodomain. We applied in situ hybridization to localize Iroquois in the sponge. The expression of this gene is highest in cells adjacent to the canals of the sponge in the medulla region. To study the expression of Iroquois during development, the in vitro primmorph system from S. domuncula was used. During the formation of these three dimensional aggregates composed of proliferating cells, the expression of Iroquois depends on ferric iron and water current. An increased expression in response to water current is paralleled with the formation of canal-like pores in the primmorphs. It is suggested that Iroquois expression is involved in the formation of the aquiferous system, the canals in sponges and the canal-like structures in primmorphs. PMID- 12752764 TI - A DM domain protein from a coral, Acropora millepora, homologous to proteins important for sex determination. AB - The identification and functional studies of DM domain-containing proteins Doublesex, MAB-3, and DMRT1 indicated that flies, nematodes, and humans share at least some of the molecular mechanisms of sex determination. We identified a gene, AmDM1, from the coral Acropora millepora that encodes a homologous DM domain-containing protein. Molecular analyses show that the AmDM1 primary transcript is processed to generate four different messenger RNAs. Alternative use of two polyadenylation sites produces transcripts that vary only in the 3' untranslated regions, whereas alternative splicing generates transcripts with and without the region coding for the DM domain. All the transcripts include a second motif, the DMA domain, which is found in a number of other proteins containing a DM domain. Hermaphroditic A. millepora differentiates sexual cells seasonally before the spring spawn, and Northern blot analysis shows that the AmDM1 transcripts are present at higher levels during sexual differentiation. The non DM domain-containing messages are also present at significant levels in late embryos, but DM domain transcripts are extremely rare at this stage. These data suggest that the association of DM domain proteins and sexual determination or differentiation predates the separation of the Cnidaria from the rest of the Metazoa. PMID- 12752765 TI - Rapid evolution of a polyphenic threshold. AB - Polyphenisms are thought to play an important role in the evolution of phenotypic diversity and the origin of morphological and behavioral novelties. However, the extent to which polyphenic developmental mechanisms evolve in natural populations is unknown. Here we contrast patterns of male phenotype expression in native and exotic and ancestral and descendant populations of the horn polyphenic beetle, Onthophagus taurus. Males in this species express two alternative morphologies in response to larval feeding conditions. Favorable conditions cause males to grow larger than a threshold body size and to develop a pair of horns on their heads. Males that encounter relatively poor conditions do not reach this threshold size and remain hornless. We show that exotic and native populations of O. taurus differ significantly in the body size threshold that separates alternative male phenotypes. Comparison with archival museum collections and additional samples obtained from the native range of O. taurus suggests that allometric differences between exotic and native populations do not reflect preexisting variation in the native range of this species. Instead, our data suggest that threshold divergences between exotic and native populations have evolved in less than 40 years since the introduction to a new habitat and have proceeded in opposite directions in two exotic ranges of this species. Finally, we show that the kind and magnitude of threshold divergence between native and exotic populations are similar to differences normally observed between species. Our results support the view that certain components of the developmental control mechanism that underlie polyphenic development can evolve rapidly in natural populations and may provide important avenues for phenotypic differentiation and diversification in nature. We discuss the role of developmental control mechanisms in the origin of allometric diversification and explore potential evolutionary mechanisms that could drive scaling relationship evolution in nature. PMID- 12752766 TI - Mimicry: developmental genes that contribute to speciation. AB - Despite renewed interest in the role of natural selection as a catalyst for the origin of species, the developmental and genetic basis of speciation remains poorly understood. Here we describe the genetics of Mullerian mimicry in Heliconius cydno and H. melpomene (Lepidoptera: Nymphalidae), sister species that recently diverged to mimic other Heliconius. This mimetic shift was a key step in their speciation, leading to pre- and postmating isolation. We identify 10 autosomal loci, half of which have major effects. At least eight appear to be homologous with genes known to control pattern differences within each species. Dominance has evolved under the influence of identifiable "modifier" loci rather than being a fixed characteristic of each locus. Epistasis is found at many levels: phenotypic interaction between specific pairs of genes, developmental canalization due to polygenic modifiers so that patterns are less sharply defined in hybrids, and overall fitness through ecological selection against nonmimetic hybrid genotypes. Most of the loci are clustered into two genomic regions or "supergenes," suggesting color pattern evolution is constrained by preexisting linked elements that may have arisen via tandem duplication rather than having been assembled by natural selection. Linkage, modifiers, and epistasis affect the strength of mimicry as a barrier to gene flow between these naturally hybridizing species and may permit introgression in genomic regions unlinked to those under disruptive selection. Mullerian mimics in Heliconius use different genetic architectures to achieve the same mimetic patterns, implying few developmental constraints. Therefore, although developmental and genomic constraints undoubtedly influence the evolutionary process, their effects are probably not strong in comparison with natural selection. PMID- 12752767 TI - A mechanistic study of evolvability using the mitogen-activated protein kinase cascade. AB - Evolvability is a function of the way genetic variation interacts with the mechanisms that produce the phenotype. We explore an explicitly mechanistic way of studying the evolvability of phenotypes that are produced by a relatively simple genetic mechanism, the mitogen-activated protein kinase (MAPK) cascade. We developed a quantitative model of MAPK activation that can be used to study the effects of genetic variation on the various components of this signaling cascade. We show how some standard tools of applied mathematics, such as steady-state formulations and nondimensionalization, can be used to elucidate the relative importance of variation in each gene of this mechanism. We also give insights into non-intuitive patterns of dependence and trade-off among the genes. The mechanism produces several different phenotypes (ultrasensitivity to stimulation, switch-like behavior, amount of MAPK-PP delivered, persistence of MAPK-PP activity), each of which is sensitive to different (but partially overlapping) combinations of genes. We show that the mechanism imposes clear limitations on the evolvability of each of the different phenotypes of the pathway, even in the presence of genetic variation in the components of the mechanism. This approach to the study of evolvability is generally applicable and complements the traditional approach through statistical genetics by providing a mechanistic understanding of the genetic interactions that produce the phenotype. PMID- 12752768 TI - Variation of tooth number in mammalian dentition: connecting genetics, development, and evolution. AB - A major question in modern biology is how gene mutations affect development and are translated into macroevolutionary changes in morphology. Variations in tooth number, a strategy used by many mammals to develop specialized dentitions, has been an important factor for species diversification. Changes in the number of teeth tend to occur in the reverse of the order teeth are formed during development, which also characterizes the general pattern of tooth loss observed during the evolution of placental mammals. To understand how changes at the molecular level affect the distinct stages of tooth development, we analyzed the ontogenesis of tooth growth arrest in sciurids and mice and in single and double knockout mutant mice. We show that the complexity of the genetic network that governs tooth development can change during ontogenetic trajectory, and these changes may be related to macroevolutionary changes. Furthermore, we show that the variation in tooth number in the affected members of human families bearing mutations in the MSX1 and PAX9 genes can help to understand how the genetic variations within a population can modulate evolutionary changes in dental patterning. PMID- 12752769 TI - Evolution of development in the cellular slime molds. PMID- 12752770 TI - Ontogenetic evidence for the Paleozoic ancestry of salamanders. AB - The phylogenetic positions of frogs, salamanders, and caecilians have been difficult to establish. Data matrices based primarily on Paleozoic taxa support a monophyletic origin of all Lissamphibia but have resulted in widely divergent hypotheses of the nature of their common ancestor. Analysis that concentrates on the character states of the stem taxa of the extant orders, in contrast, suggests a polyphyletic origin from divergent Paleozoic clades. Comparison of patterns of larval development in Paleozoic and modern amphibians provides a means to test previous phylogenies based primarily on adult characteristics. This proves to be highly informative in the case of the origin of salamanders. Putative ancestors of salamanders are recognized from the Permo-Carboniferous boundary of Germany on the basis of ontogenetic changes observed in fossil remains of larval growth series. The entire developmental sequence from hatching to metamorphosis is revealed in an assemblage of over 600 specimens from a single locality, all belonging to the genus Apateon. Apateon forms the most speciose genus of the neotenic temnospondyl family Branchiosauridae. The sequence of ossification of individual bones and the changing configuration of the skull closely parallel those observed in the development of primitive living salamanders. These fossils provide a model of how derived features of the salamander skull may have evolved in the context of feeding specializations that appeared in early larval stages of members of the Branchiosauridae. Larvae of Apateon share many unique derived characters with salamanders of the families Hynobiidae, Salamandridae, and Ambystomatidae, which have not been recognized in any other group of Paleozoic amphibians. PMID- 12752772 TI - Neurokinin 1 receptor and relative abundance of the short and long isoforms in the human brain. AB - Substance P exerts its various biochemical effects mainly via interactions through neurokinin-1 receptors (NK1). Recently, the NK1 receptor has attracted considerable interest for its possible role in a variety of psychiatric disorders including depression and anxiety. However, little is known regarding the anatomical distribution of NK1 in the human central nervous system (CNS). Riboprobe in situ hybridization, quantitative PCR and in vitro autoradiography were performed. Highest NK1 mRNA levels were localized in the locus coeruleus and ventral striatum, while moderate hybridization signals were observed in the cerebral cortex (most abundant in the visual cortex), hippocampus and different amygdaloid nuclei. Very low levels of the NK1 mRNA were detected in the cerebellum and thalamus. In view of the existence of a long and short isoform of the NK1 receptor, it was of interest to assess whether there was a differential distribution of the two splice variants in the human CNS and peripheral tissues. A quantitative TaqMan PCR analysis showed that the long NK1 isoform was the most prevalent throughout the human brain, while in peripheral tissues the truncated form was the most represented. 3H-Substance P autoradiography revealed a good correlation between receptor binding sites and NK1 mRNA expression throughout the brain, with the highest levels of binding in the locus coeruleus. These results provide the anatomical evidence that the NK1 receptors have a strong association with neuronal systems relevant to mood regulation and stress in the human brain, but do not suggest a region-specific role of the two isoforms in the CNS. PMID- 12752771 TI - Intrinsic light responses of retinal ganglion cells projecting to the circadian system. AB - In mammals, light entrainment of the circadian clock, located in the suprachiasmatic nuclei (SCN), requires retinal input. Traditional rod and cone photoreceptors, however, are not required. Instead, the SCN-projecting retinal ganglion cells (RGCs) function as autonomous photoreceptors and exhibit light responses independent of rod- and cone-driven input. Using whole-cell patch-clamp recording techniques, we have investigated the morphological and electrophysiological properties of this unique class of RGCs. Although SCN projecting RGCs resemble Type III cells in form, they display strikingly different physiological properties from these neurons. First, in response to the injection of a sustained depolarizing current, SCN-projecting cells fired in a transient fashion, in contrast to most RGCs which fired robust trains of action potentials. Second, in response to light, SCN-projecting RGCs exhibited an intensity-dependent transient depolarization in the absence of rod and cone input. This depolarization reached a peak within 5 s and generated increased spiking activity before decaying to a plateau. Voltage-clamp recordings were used to characterize the light-activated conductance which generated this depolarization. In response to varying light intensities, SCN-projecting RGCs exhibited a graded transient inward current which peaked within 5 s and decayed to a plateau. The voltage dependence of the light-activated current was obtained by subtracting currents elicited by a voltage ramp before and during illumination. The light-activated current displayed both inward and outward rectification and was largely unaffected by substitution of extracellular Na+ with choline. In both respects, the intrinsic light-activated current observed in SCN-projecting RGCs resembles currents carried by ion channels of the transient receptor potential (trp) family, which are known to mediate the light response of invertebrate photoreceptors. PMID- 12752773 TI - Ontogenetic development of cannabinoid receptor expression and signal transduction functionality in the human brain. AB - Previous evidence suggests that the endogenous cannabinoid system emerges relatively early during brain development in the rat. However, the pre- and postnatal pattern of appearance of CB1 cannabinoid receptors in humans has not been analysed in detail. Furthermore, there is a complete lack of information about the functional ability of these proteins to activate signal transduction mechanisms during human development. In the present study we have explored CB1 receptor expression throughout the different areas of the developing human brain by [3H]CP55 940 autoradiography. We have also assessed CB1 functional coupling to G proteins during brain development by agonist-stimulated [35S]GTPgammaS autoradiography in the same cases. Our results indicate a significant density of cannabinoid receptors at 19 weeks' gestation in the same areas that contain these receptors in the adult human brain. Autoradiographic levels of CB1 receptors in these structures seem to increase progressively from early prenatal stages to adulthood. Interestingly, high densities of cannabinoid receptors have also been detected during prenatal development in fibre-enriched areas that are practically devoid of them in the adult brain. In parallel with these data, we have found that brain cannabinoid receptors are functionally coupled to signal transduction mechanisms from early prenatal stages. This early pattern of expression of functionally active cannabinoid receptors, along with the transient and atypical localization of these proteins in white matter areas during the prenatal stages, suggest an specific role of the endocannabinoid system in the events related to human neural development. PMID- 12752774 TI - Enzymatic removal of chondroitin sulphates abolishes the age-related axon order in the optic tract of mouse embryos. AB - Retinal axons undergo an age-related reorganization at the junction of the chiasm and the optic tract. We have investigated the effects of removal of chondroitin sulphate on this order change in mouse embryos aged embryonic day 14, when most axons are growing in the optic tract. Enzymatic removal of chondroitin sulphate but not keratan sulphate in brain slice preparations of the retinofugal pathway abolished the accumulation of phalloidin-positive growth cones in the subpial region of the optic tract. The loss of chronotopicity was further demonstrated by anterograde filling of single retinal axons, which showed a dispersion of growth cones from subpial to the whole depth of the tract. The enzyme treatment neither produced detectable changes in growth cone morphology and growth dynamic of retinal neurites nor affected the radial glial processes in the tract, indicating a specific effect of removal of chondroitin sulphate from the pathway to the axon order in the tract. Although chondroitin sulphate was also found at the midline of the chiasm, growth cone distribution across the depth of fibre layer at the midline was not affected by the enzyme treatment. These results suggest a mechanism in which retinal axons undergo changes in response to chondroitin sulphate at the chiasm-tract junction, but not at the midline, that produce a chronotopic fibre rearrangement in the mouse retinofugal pathway. PMID- 12752775 TI - Gamma-frequency fluctuations of the membrane potential and response selectivity in visual cortical neurons. AB - Fluctuations at frequencies of 25-70 Hz is an inherent property of cortical activity. These rapid, gamma-range fluctuations are apparent in the local field potentials, in spiking of cells and cell groups, and in the membrane potential of neurons. To investigate stimulus dependence of the gamma-frequency fluctuations of the membrane potential, we have recorded intracellularly responses of cells in cat visual cortex to presentation of moving gratings. We found gamma-range fluctuations of the membrane potential in both simple and complex cells. The strength of the gamma-frequency fluctuations correlated with the stimulus optimality. Furthermore, the amplitude of the gamma-frequency fluctuations correlated with the phase of stimulus-imposed slow changes of the membrane potential. The combination of these features makes cortical neurons capable of encoding the slow changes in the visual world in a kind of amplitude modulation of the high frequency fluctuations. This assures reliable transformation of the membrane potential changes into spike responses without compromising the temporal resolution of visual information encoding in the low frequency range. PMID- 12752776 TI - Modulation of GABAergic transmission by endogenous glutamate in the rat supraoptic nucleus. AB - The presence of group III metabotropic glutamate receptors on GABAergic terminals in the supraoptic nucleus suggests that the level of glutamate in the extracellular space may regulate synaptic strength at inhibitory synapses. To test this hypothesis we examined the consequences of increasing ambient glutamate on GABA-mediated synaptic activity in supraoptic neurons. The concentration of the excitatory amino acid in the extracellular space was increased pharmacologically by blocking glutamate transporters. Inhibition of the astrocyte specific GLT-1 glutamate transporter led to a reversible decrease in evoked inhibitory postsynaptic current amplitude. This modulation had a presynaptic origin as revealed by analysis of paired-pulse ratio and miniature inhibitory currents. Furthermore, blocking group III metabotropic glutamate receptors with the specific antagonist MAP4 prevented the depression of GABAergic transmission induced by glutamate transporter blockade. Thus, presynaptic metabotropic glutamate receptors located on inhibitory terminals in the supraoptic nucleus appear to sense changes in ambient glutamate and modify GABA release accordingly. However, it seems that such changes need to reach a certain magnitude because the discrete deficit in glutamate clearance which occurs in the supraoptic nucleus of lactating rats is not sufficient to modulate GABA-mediated transmission. These results suggest that ambient glutamate contributes to the modulation of synaptic efficacy not only at glutamatergic synapses but also at inhibitory GABAergic synapses. PMID- 12752777 TI - Evidence for a function-specific mutation in the neurotoxin, parabutoxin 3. AB - Parabutoxin 3 (PBTx3), a short-chain alpha-K+ neurotoxin from the scorpion, Parabuthus transvaalicus, is a 37-residue polypeptide cross-linked by three disulphide bridges. The affinity towards Kv1 channels is very weak (Kd approximately 79 micro m for Kv1.1 channels), or moderate (Kd approximately 500 nm for Kv1.2 and Kv1.3 channels). In an effort to generate a more potent K+ channel blocker, we recombinantly produced a mutant PBTx3 by the introduction of an aromatic amino acid, fenylalanine in close proximity of the crucial lysine 26 residue, to create a functional diad similar to subfamily three alpha-K+ toxins. The mutant was tested for his ability to block Kv1.1, Kv1.2 and Kv1.3 channels in Xenopus laevis oocytes: a hundred-fold higher affinity towards Kv1.1 channels and a fivefold increase in affinity towards Kv1.3 channels was observed, when compared to the wild-type toxin. The effect on Kv1.2 channels was similar to the wild-type toxin, indicating a specific interaction site for the mutated residue onto the different Kv-type channels. PMID- 12752778 TI - Melanopsin retinal ganglion cells and the maintenance of circadian and pupillary responses to light in aged rodless/coneless (rd/rd cl) mice. AB - Melanopsin-expressing ganglion cells have been proposed as the photoreceptors mediating non-rod, non-cone ocular responses to light. Here we use the aged (approximately 2 years) rodless and coneless (rd/rd cl) mouse to assess the impact of progressive inner retinal cell loss on melanopsin expression, circadian entrainment and pupillary constriction. Aged rd/rd cl mice show substantial transneuronal retinal degeneration leaving only the ganglion cell layer and little of the inner nuclear layer. Despite this loss, quantitative reverse transcriptase-polymerase chain reaction showed normal levels of melanopsin expression, and immunocytochemistry demonstrated both the presence and normal cellular appearance of these cells. Furthermore, the optic nerves of the two genotypes (rd/rd cl and +/+) were not obviously different in animals older than 2 years. However, this massive level of retinal degeneration left both pupillary and circadian responses to light intact, even in rd/rd cl mice older than 2 years. Our data provide the first positive correlation between the persistence of melanopsin-expressing cells and the maintenance of both circadian and pupillary responses to light in the absence of rods and cones. These findings, together with recent studies on melanopsin knockout mice, are consistent with the hypothesis that melanopsin-expressing ganglion cells are photosensitive and mediate a range of irradiance-detection tasks. PMID- 12752779 TI - Regulation of alpha1G T-type calcium channel gene (CACNA1G) expression during neuronal differentiation. AB - Down-regulation of T-type Ca channel current and mRNA occurs following differentiation of Y79 retinoblastoma cells. To understand how the decrease in expression is linked to cell differentiation, we examined transcriptional regulation of the Cav3.1 Ca channel gene, CACNA1G. We identified two putative promoters (A and B) in 1.3 kb of cloned genomic DNA. Reverse transcriptase polymerase chain reaction and 5' rapid amplification of cDNA ends-polymerase chain reaction analyses demonstrated that two transcripts with different 5' untranslated regions are generated by different transcription start sites, with promoter A favoured in undifferentiated cells and promoter B favoured in differentiated cells. Functional analyses of the promoter sequence revealed that both promoters are active. Enhancer and repressor sequences were identified upstream of promoter A and B, respectively. These results suggest that the down regulation of alpha1G mRNA in differentiated Y79 cells is mediated primarily by decreased activity of promoter A, which could occur in conjunction with repression of the activity of promoter B. The decrease in T-type Ca channel expression in Y79 cells may be an essential signal affecting phenotypic maturation and expression of other ion channel subtypes in the differentiated cells. PMID- 12752780 TI - Rhythm sequence through the olfactory bulb layers during the time window of a respiratory cycle. AB - The mammalian olfactory bulb is characterized by prominent oscillatory activity of its local field potentials. Breathing imposes the most important rhythm. Other rhythms have been described in the beta- and gamma-frequency ranges. We recorded unitary activities in different bulbar layers simultaneously with local field potentials in order to examine the different relationships existing between (i) breathing and field potential oscillations, and (ii) breathing and spiking activity of different cell types. We show that, whatever the layer, odour-induced gamma oscillations always occur around the transition point between inhalation and exhalation while beta oscillations appear during early exhalation and may extend up to the end of inhalation. By contrast, unitary activities exhibit different characteristics according to the layer. They vary in (i) their temporal relationship with respect to the respiratory cycle; (ii) their spike rates; (iii) their temporal patterns defined according to the respiratory cycle. The time window of a respiratory cycle might thus be split into three main epochs based on the deceleration of field potential rhythms (from gamma to beta oscillations) and a simultaneous gradient of spike discharge frequencies ranging from 180 to 30 Hz. We discuss the possibility that each rhythm could serve different functions as priming, gating or tuning for the bulbar network. PMID- 12752781 TI - Impaired glutamate uptake and EAAT2 downregulation in an enterovirus chronically infected human glial cell line. AB - Rapid and efficient uptake of glutamate via the high-affinity glutamate transporter EAAT2 is important for limiting glutamate-mediated excitotoxicity involved in neuronal death. Furthermore, there is evidence of altered glutamate uptake and catabolism in motor neuron diseases. Such a defect has been reported in amyotrophic lateral sclerosis, the major motor neuron disease, and was associated with impairment in EAAT2 processing. We recently reported the presence of enterovirus genome specifically in the anterior horn of amyotrophic lateral sclerosis cases, suggesting the involvement of a chronic/persistent enterovirus infection in amyotrophic lateral sclerosis. To investigate a putative link between enterovirus infection and the glutamate-mediated excitotoxicity observed in amyotrophic lateral sclerosis, we developed an in vitro model consisting of a human glial cell line infected with ECHOvirus 6, one of the enteroviruses with sequences closely related to those detected in patients with amyotrophic lateral sclerosis. In these glial cells, an ECHOvirus 6 chronic infection was established, resulting in altered extracellular glutamate uptake. This correlated with an aberrant splicing of the EAAT2 pre-messenger ribonucleic acid and a significant loss of EAAT2 protein expression, similar to that observed in patients with amyotrophic lateral sclerosis. These results provide convincing evidence that an enterovirus chronic/persistent infection may alter glial glutamate uptake and catabolism. As enteroviruses are extremely common human pathogens, they may act as a trigger in the development of certain motor neuron diseases, such as amyotrophic lateral sclerosis. PMID- 12752782 TI - Reduced spinal cord sensitization to C-fibre stimulation in mice over-expressing galanin. AB - The neuropeptide galanin may have a role in spinal nociception. In this study, we examined the excitability of the flexor reflex and its sensitization by repetitive stimulation of nociceptive C-fibres in anaesthetized mice that over express galanin. No difference was seen between over-expressing galanin and wild type mice in the magnitude of the baseline flexor reflex. Repetitive conditioning stimulation of C-fibres (10 stimuli at 1 Hz) produced a gradual increase in reflex magnitude during the conditioning stimulation (wind-up), as well as an increase in spinal reflex excitability after the termination of the stimulus train (central sensitization) in wild-type mice. Although the wind-up did not differ between over-expressing galanin and wild-type mice, the magnitude of central sensitization was significantly reduced in the over-expressing galanin mice (24 +/- 13% peak increase compared with 164 +/- 65% in the wild-type). Intrathecal administration of M35, a galanin receptor antagonist, markedly enhanced central sensitization in over-expressing galanin mice in association with C-fibre conditioning stimulation, while having no effect in wild-type mice. These results provide further electrophysiological evidence for an inhibitory function of galanin in modulation of central sensitization in response to C-fibre stimulation. PMID- 12752783 TI - Presence of functional vasopressin receptors in spinal ventral horn neurons of young rats: a morphological and electrophysiological study. AB - The objective of the present work was double. (i) Light microscopic autoradiography was used to determine the distribution of vasopressin and oxytocin binding sites in the spinal cord of rats. (ii) Whole-cell recordings were performed in lumbar spinal cord slices in order to assess whether these receptors are functional, whether they are located pre- or postsynaptically and whether they are present in motoneurons. In newborns, vasopressin binding sites of the V1a type were present in all laminae of the central gray at all segmental levels, whereas oxytocin binding sites were found only in the superficial layers of the dorsal horn. In adults, binding sites for both neuropeptides were also present, but were less dense. The dissociation constants for vasopressin were similar in newborns and adults. Whole-cell recordings showed that in identified motoneurons vasopressin exerted a direct effect, by inducing a membrane depolarization or by generating a sustained inward current, and an indirect effect, by enhancing glycinergic and GABAergic inhibitory transmission. Vasopressin-induced facilitation of inhibitory transmission could also be demonstrated in unidentified ventral horn neurons. All these effects were mediated by V1a but not V1b receptors. In some neurons, glycinergic transmission was also facilitated by a selective oxytocin receptor agonist. Our data, together with data obtained previously in brainstem motor nuclei, suggest that vasopressin of hypothalamic origin could play a role in motricity. The neuropeptide could act as a neuromodulator, because it would not directly activate motoneurons, but rather render them more responsive to incoming excitatory inputs. Vasopressin may thus act as a regulator of muscular force. PMID- 12752784 TI - Tumor necrosis factor receptor type-1 in sensory neurons contributes to induction of chronic enhancement of inflammatory hyperalgesia in rat. AB - Carrageenan-induced inflammatory pain lasting hours to days produces a protein kinase C epsilon (PKC epsilon )-dependent 'primed' state lasting several weeks, during which time injection of prostaglandin E2 induces hyperalgesia which is markedly enhanced and prolonged compared to PGE2-induced hyperalgesia in normal 'unprimed' rats. In the present study, we demonstrate that while inhibition of prostaglandin synthesis and antagonism of beta2-adrenergic receptors markedly attenuated the hyperalgesia induced by carrageenan, these interventions did not affect hyperalgesic priming. Tumor necrosis factor-alpha (rat recombinant; rrTNFalpha), another mediator of carrageenan-induced inflammation, alone produced hyperalgesia and priming, which were attenuated and prevented, respectively, by intrathecal administration of antisense to PKC epsilon. Inhibition of TNFalpha with thalidomide or a rat polyclonal anti-TNFalpha antibody attenuated carrageenan-induced hyperalgesia and prevented priming. Intrathecal administration of antisense to tumour necrosis factor receptor type-1 (TNFR1) reduced the level of TNFR1 transported toward the peripheral terminals of sensory neurons, and attenuated both carrageenan- and rrTNFalpha-induced priming. Acute hyperalgesia induced by carrageenan or rrTNFalpha remained intact in animals treated with TNFR1 antisense. Our results demonstrate that the generation of the primed state does not require production of hyperalgesia and that TNFalpha, which is generated during acute inflammation, can act on sensory neurons to induce hyperalgesic priming by activating neuronal PKC epsilon. PMID- 12752785 TI - Expression of neuroserpin in the visual cortex of the mouse during the developmental critical period. AB - The neuronal serine protease inhibitor neuroserpin is widely expressed in the developing and adult brain. In the neocortex, neuroserpin is displayed particularly during the period of synaptic specification and refinement, indicating a role as modulator of extracellular proteolytic processes. The synaptic connections of the visual system of the mouse are shaped during early postnatal life by an activity-dependent process. We have studied the expression of the neuronal serine protease inhibitor neuroserpin in the primary visual cortex of mice from birth until the end of the critical period by means of reverse transcription polymerase chain reaction and in situ hybridization. The localization and the level of expression were constant throughout this period. Monocular deprivation with an eyelid sutured induced a decrease in neuroserpin expression in neurons of area 17 after 1 week of deprivation, the decrease being more pronounced on the side contralateral to the closed eye. The expression of neuroserpin in the visual cortex during the critical period and its decrease in parallel to the refinement of synaptic contacts after visual deprivation suggests a regulative role of neuroserpin on these processes. PMID- 12752786 TI - Interneurons are the local targets of hippocampal inhibitory cells which project to the medial septum. AB - A subset of GABAergic neurons projecting to the medial septum has long been described in the hippocampus. However, the lack of information about their local connectivity pattern or their correspondence with any of the well-established hippocampal interneuron types has hampered the understanding of their functional role. Retrograde tracing combined with immunostaining for neurochemical markers in the adult rat hippocampus showed that nearly all hippocampo-septal (HS) neurons express somatostatin (>95%) and, in the hilus and CA3 stratum lucidum, many contain calretinin (>45%). In contrast, in stratum oriens of the CA1 and CA3 subfields, the majority of HS neurons contain somatostatin (>86%) and calbindin (>73%), but not calretinin. Because somatostatin-positive hippocampal interneurons have been most extensively characterized in the stratum oriens of CA1, we focused our further analysis on HS cells found in this region. In 18-20 day-old rats, intracellularly filled CA1-HS cells had extensive local axon collaterals crossing subfield boundaries and innervating the CA3 region and the dentate gyrus. Electron microscopic analysis provided evidence that the axon terminals of CA1-HS cells form symmetrical synapses selectively on GABAergic interneurons, both locally and in the CA3 region. In addition, double retrograde labelling experiments revealed that many CA1-HS neurons of the dorsal hippocampus also have collateral projections to the ventral hippocampus. Thus, CA1-HS cells innervate inhibitory interneurons locally and in remote hippocampal regions, in addition to targeting mostly GABAergic neurons in the medial septum. This dual projection with striking target selectivity for GABAergic neurons may be ideally suited to synchronize neuronal activity along the septo-hippocampal axis. PMID- 12752787 TI - Early establishment of multiple release site connectivity between interneurons and pyramidal neurons in the developing hippocampus. AB - The strength of the synaptic transmission between two neurons critically depends on the number of release sites connecting the neurons. Here we examine the development of connectivity between gamma-aminobutyric acid (GABA)ergic interneurons and CA1 pyramidal neurons in the hippocampus. GABAergic postsynaptic currents (PSCs) were recorded in whole-cell voltage-clamped CA1 pyramidal neurons. By comparing spontaneous and miniature (action potential-independent) GABAergic PSCs, we found that multiple release site connectivity is established already at the first postnatal day and that the degree of connectivity remains unaltered into adulthood. During the same time there is a dramatic increase in the number of GABAergic synapses on each pyramidal neuron as indicated by the increase in frequency of miniature GABAergic PSCs. These results indicate that during development a given interneuron contacts an increasing number of target pyramidal neurons but with the same multiple release site connectivity. It has been shown previously that the connectivity between CA3 and CA1 pyramidal neurons is initially restricted to one release site, and develops gradually. The present result thus suggests different mechanisms to govern the maturation of excitatory and inhibitory synaptic transmissions. PMID- 12752788 TI - Somatostatin receptor 2 knockout/lacZ knockin mice show impaired motor coordination and reveal sites of somatostatin action within the striatum. AB - The peptide somatostatin can modulate the functional output of the basal ganglia. The exact sites and mechanisms of this action, however, are poorly understood, and the physiological context in which somatostatin acts is unknown. Somatostatin acts as a neuromodulator via a family of five 7-transmembrane G protein-coupled receptors, SSTR1-5, one of which, SSTR2, is known to be functional in the striatum. We have investigated the role of SSTR2 in basal ganglia function using mice in which Sstr2 has been inactivated and replaced by the lacZ reporter gene. Analysis of Sstr2lacZ expression in the brain by beta-galactosidase histochemistry demonstrated a widespread pattern of expression. By comparison to previously published in situ hybridization and immunohistochemical data, Sstr2lacZ expression was shown to accurately recapitulate that of Sstr2 and thus provided a highly sensitive model to investigate cell-type-specific expression of Sstr2. In the striatum, Sstr2 expression was identified in medium spiny projection neurons restricted to the matrix compartment and in cholinergic interneurons. Sstr2 expression was not detected in any other nuclei of the basal ganglia except for a sparse number of nondopaminergic neurons in the substantia nigra. Microdialysis in the striatum showed Sstr2-null mice were selectively refractory to somatostatin-induced dopamine and glutamate release. In behavioural tests, Sstr2-null mice showed normal levels of locomotor activity and normal coordination in undemanding tasks. However, in beam-walking, a test of fine motor control, Sstr2-null mice were severely impaired. Together these data implicate an important neuromodulatory role for SSTR2 in the striatum. PMID- 12752789 TI - Effect of sleep and sleep deprivation on serotonergic neurotransmission in the hippocampus: a combined in vivo microdialysis/EEG study in rats. AB - Brainstem serotonergic neurotransmission is implicated in sleep regulation. However, the role of serotonin (5-HT) in forebrain regions in sleep-wake mechanisms is still unclear. Here, we have investigated, using a combined in vivo microdialysis/electroencephalogram method, the relationship between hippocampal 5 HT levels and sleep-wake behaviour in the rat. A clear-cut relationship was found between hippocampal 5-HT levels and vigilance state. The highest levels of 5-HT were observed during wakefulness, whereas a progressive decrease of 5-HT going from nonrapid eye movement sleep to rapid eye movement sleep was found. Sleep deprivation (SD) causes a transient enhancement of mood in depressed patients. Given the putative role of 5-HT in the aetiology of depression and the therapeutical efficacy of selective serotonin reuptake inhibitors in this illness, we also studied hippocampal 5-HT during 4 h of SD and during the subsequent recovery period. During the whole SD period, 5-HT levels were elevated substantially when compared to 5-HT levels during basal wakefulness. However, no changes in 5-HT levels and the relationship between hippocampal 5-HT and vigilance state were found during the subsequent recovery period. As SD is a potentially stressful experience and glucocorticoids are involved in the regulation of serotonergic neurotransmission and sleep, we investigated the effects of SD on free corticosterone levels. SD caused a marked rise in free corticosterone levels. However, the effects of SD on 5-HT seem not to be mediated by this hormone, because adrenalectomy did not affect the rise in hippocampal 5 HT during SD. We hypothesize that the elevated hippocampal 5-HT levels during SD may participate in the transient mood enhancing properties of forced wakefulness observed in depressed patients. PMID- 12752790 TI - A subpopulation of rats show social and sleep-waking changes typical of chronic neuropathic pain following peripheral nerve injury. AB - Neuropathic conditions for which treatment is sought, the so-called chronic pain syndrome, are characterized usually by complex behavioural disturbances as well as pain. In this study we evaluated whether social behavioural and sleep disruptions occurred after nerve injury. Before and after chronic constriction of the sciatic nerve, resident-intruder and sleep-wake cycles, as well as mechanical and thermal allodynia/hyperalgesia, were quantified. Sciatic nerve injury in all animals reduced withdrawal thresholds to tactile and thermal (cold) stimuli. Resident-intruder and sleep-waking behaviours were altered in some but not all animals. One group (30%, 'persistent change') had enduring reductions in dominant behaviour to an intruder and decreased slow-wave sleep and increased wakefulness during both light and dark cycles. Another group (25%, 'recovery') had a transient reduction in dominant behaviours and decreased slow-wave sleep and increased wakefulness during only the light cycle. In a third group (45%, 'no effect') resident-intruder and sleep-waking behaviours remained normal. Our finding that the degree of 'pain' as inferred from the allodynia/hyperalgesia was identical in all animals suggests that the alterations to resident-intruder and sleep-wake cycles were independent of the level of sensory disturbance. An absence of correlation between intensity of sensory disturbances and measures of disability (loss of sleep, familial/social problems) is also characteristic of human neuropathic pain. These data indicate that: (i) in a subpopulation of animals sciatic injury results in two of the major complex behavioural changes which are characteristic of neuropathic pain in humans; (ii) testing only for allodynia and hyperalgesia is not sufficient to detect this subpopulation. PMID- 12752791 TI - Learning deficits and dysfunctional synaptic plasticity induced by aggregated amyloid deposits in the dentate gyrus are rescued by chronic treatment with indomethacin. AB - The amyloid pathology in Alzheimer's disease is accompanied by a chronic inflammatory response characterized by gliosis and activated microglial cells surrounding senile plaques. Epidemiological studies have shown nonsteroidal anti inflammatory drug treatment reduces the risk of Alzheimer's disease. We have previously shown that injection of a combination of Abeta40 and Abeta43 in the dentate gyrus of the rat induces aggregated amyloid deposits and inflammation associated with dysfunctional synaptic plasticity and learning deficits. Here we characterize the effectiveness of nonsteroidal anti-inflammatory treatment in this model and show that this treatment restores the working memory deficit and decremental long-term potentiation in the dentate gyrus. Importantly, we observe no qualitative difference in the presence of aggregated material but a substantial reduction in microglial-induced inflammation, suggesting that mature aggregated plaques may not be directly responsible for the deficits but may trigger an inflammatory response which has a detrimental effect on synaptic function and memory. PMID- 12752792 TI - Chronic unpredictable stress impairs long-term potentiation in rat hippocampal CA1 area and dentate gyrus in vitro. AB - Rises in corticosteroid levels, e.g. after acute stress, impair synaptic plasticity in the rat hippocampus when compared with the situation where levels are basal, i.e. under rest. We here addressed the question whether basal and raised levels of corticosterone affect synaptic plasticity similarly in animals that experienced chronic stress prior to corticosterone application. To this end, rats were exposed to a 21-day variable stress paradigm. Synaptic plasticity was examined in vitro in the dentate gyrus and CA1 hippocampal region, 24 h after exposure to the last stressor, i.e. when corticosterone levels are basal (low). First we observed that long-term potentiation was greatly impaired in both CA1 and dentate gyrus after 3 weeks of exposure to variable stress, when recorded under conditions where plasma corticosterone levels are low. Second, administration of 100 nm corticosterone in vitro reduced synaptic plasticity in CA1 of control rats, but induced no further impairment of synaptic plasticity in chronically stressed rats. Third, in the dentate gyrus, corticosterone incubation did not affect synaptic plasticity in slices from both control and stressed animals. We conclude that: (i) exposure to chronic variable stress per se reduces synaptic plasticity both in CA1 and dentate gyrus; and (ii) acute rises in corticosterone level induce no additional impairment of synaptic plasticity in the CA1 region of chronically stressed rats. It is tempting to speculate that the stress-induced reduction of hippocampal efficacy provides a cellular substrate for cognitive deficits in hippocampus-dependent learning tasks seen after prolonged exposure to stressful events. PMID- 12752793 TI - Neural correlates of memorized associations and cued movements in archistriatum of the domestic chick. AB - The archistriatum mediates a neural pathway from the medial part of intermediate hyperstriatum ventrale (in the dorsal pallium) to the lobus parolfactorius (in the medial striatum), thus is possibly involved in memory formation in the domestic chick. To elucidate the functional roles, we examined single neuron activities from archistriatum in unconstrained chicks during execution of a GO/NOGO task. In this task, a brief motor sound was given as initial cue, and immediately followed by presentation of a coloured bead. Chick was required to recall the memorized associations between the colour and reward, and pecked at the bead to gain food after a delay (GO trials) or stayed not pecking (NOGO trials). The ventral part of intermediate archistriatum proved to contain a group of neurons that selectively responded to the reward-associated colours before the reward was actually presented, possibly coding the memorized associations. Another group of neurons fired during the reward period, thus could code aspects of the food reward. Yet another group of neurons started to fire immediately on the cue sound and prior to the cued movements nonselectively in both GO and NOGO trials, thus could be involved in the sensori-motor link between the sound and the targeted body movements. It is concluded that even a subregion of archistriatum contains diverse neural codes for memorized associations and food rewards, and neural codes of movements cued by sounds, suggesting that archistriatum is a complex of different functional systems, possibly corresponding to striatum, limbic amygdala, and prefrontal cortex in mammals. PMID- 12752794 TI - Deficit in long-term contextual fear memory induced by blockade of beta adrenoceptors in hippocampal CA1 region. AB - The present study investigated the effects of intra-CA1 infusion of d,l propranolol, the beta-adrenergic antagonist, on memory for contextual fear conditioning. d,l-Propranolol administered 5 min ('0 h') postconditioning impaired long-term but not short-term contextual fear memory, while it was ineffective when administered 6 h postconditioning, suggesting that there is a time window for beta-adrenoceptors to play a role. Thus, we conclude that beta adrenoceptors in area CA1 are involved in regulating consolidation of contextual fear memory, with '0 h' but not 6 h post-training, a sensitive time point for the beta-adrenergic involvement. PMID- 12752795 TI - The effects of ibotenic hippocampal lesions on discriminative fear conditioning to context in mice: impairment or facilitation depending on the associative value of a phasic explicit cue. AB - To what extent the hippocampus is required for contextual conditioning remains a matter of debate. The present experiments examined the effects of ibotenate hippocampal lesions on discriminative fear conditioning to context in mice using measures of freezing in two conditioning paradigms. In both paradigms animals received foot shock as the unconditional stimulus (US) when placed in the (conditioning) context and no foot-shock when placed in the other (neutral) context. In both contexts, animals were presented with a tone as the conditioned stimulus (CS). In the conditioning context there was either no interval (delay condition) or a 30-s interval (trace condition) between tone CS end and shock US onset. These two paradigms were used because theory predicts that in the trace condition animals would learn more about contextual cues as predictors, or not, of shock US occurrence than in the delay condition. In agreement with this, we observed that sham-operated mice learned the context discrimination faster in the trace than in the delay condition. Lesions of the hippocampus significantly retarded, but did not prevent, the acquisition of the context discrimination in the trace condition. In contrast, lesions produced an opposite (facilitatory) effect in the delay condition, which was mainly observed during tone CS presentation. The data suggest that mice used two distinct competing strategies in solving this discrimination task: (i) a strategy relying on the processing of background contextual stimuli allowing direct establishment of context-US associations of different strengths, and (ii) a conditional cue (tone)-based strategy. Hence, hippocampal lesions may impair the use of the former strategy while exacerbating (unmasking) the use of the latter. PMID- 12752796 TI - Induction of the learning and plasticity-associated gene Zif268 following exposure to a discrete cocaine-associated stimulus. AB - We investigated whether the expression of the plasticity-associated gene, zif268, was associated with memories retrieved by exposure to a discrete stimulus that had been associated with cocaine, either self-administered or administered noncontingently. In the absence of drug, passive presentation of a cocaine associated light stimulus induced changes in the expression of zif268 measured by in situ hybridization within a limbic cortical-ventral striatal circuit that was not only regionally selective but related to whether the rats had originally received response-contingent or noncontingent stimulus-drug pairings. In rats that had self-administered drug, the cocaine-conditioned stimulus (CS) increased zif268 expression in neurons of the ventral tegmental area, nucleus accumbens core and shell, and basal nucleus of the amygdala but not hippocampus, prelimbic area of the medial prefrontal cortex or amygdala central nucleus. The same CS that had been associated with cocaine administered noncontingently additionally increased zif268 mRNA levels in area Cg1 of the anterior cingulate cortex, ventral and lateral regions of the orbitofrontal cortex and lateral nucleus of the amygdala. Zif268 induction was related to the predictive relationship between the stimulus and cocaine as no changes were seen in cocaine-experienced rats that had received unpaired light and drug presentations during training. Thus, zif268 expression is increased by appetitively (drug) conditioned stimuli after Pavlovian learning. Zif268 may participate in the molecular mechanisms underlying the reconsolidation or re-encoding of Pavlovian stimulus-drug associations across a distributed limbic cortical-ventral striatal neural network and that may contribute to the basis of the enduring drug-seeking behaviour produced by environmental cues. PMID- 12752797 TI - Morphological changes in hippocampal dentate gyrus synapses following spatial learning in rats are transient. AB - The hippocampus is believed to play a crucial role in the formation of memory for spatial tasks. In the present study quantitative electron microscopy was used to investigate morphological changes in the hippocampal dentate gyrus of 3-month-old male rats at 3, 9 and 24 h after training to find a hidden platform in a Morris water maze. Average escape latency (time taken to reach the platform) in all trained groups decreased progressively with increased training but data from a probe trial (quadrant analysis test) at the end of training indicated that only animals in the 9- and 24-h groups, not the 3-h group, displayed significant retention of platform location. Unbiased stereological methods were used to estimate synapse and neuronal density at each time point after training. The majority of synapses had unperforated postsynaptic densities, were localized on small dendritic spines and were classed as axo-spinous. In comparison to age matched untrained rats, significant but transient increases were observed in axo spinous synapse density and synapse-to-neuron ratio 9 h after the start of training, but not at earlier (3 h) or later (24 h) times. These changes at 9 h post-training were accompanied by transient decreases in both mean synaptic height and area of postsynaptic density. No such changes were observed in an exercise-matched control group of rats, indicating that the transient synaptic changes in the dentate gyrus are most likely to be specifically related to processes involved in memory formation for the spatial learning task. PMID- 12752798 TI - Interhemispheric visuo-motor integration in humans: the effect of redundant targets. AB - We used event-related functional magnetic resonance imaging (fMRI) to investigate the functional locus of response facilitation during parallel visuo-motor processing. In a simple reaction-time task, subjects typically respond faster to two copies of the same stimulus than to a single copy. This facilitation, called the redundant-target effect, can occur at three functional levels: perceptual, 'cognitive' or motor. Normal right handers were studied while performing a simple reaction-time task to unilateral (left or right) and bilateral light flashes. Subjects were instructed to respond with their right index finger. Reaction times were faster to bilateral light flashes than to unilateral ones, even right flashes. Greater fMRI signal for bilateral stimuli compared to unilateral ones was observed in the left precentral and postcentral gyrus, and in the right precentral gyrus. A greater fMRI signal for bilateral and for unilateral left stimuli, compared to unilateral right stimuli, was observed in an area of the right intraparietal sulcus. These results support the hypothesis that the functional locus of response facilitation during parallel visuo-motor processing is premotor. PMID- 12752799 TI - Blockade of nociceptin/orphanin FQ-NOP receptor signalling produces antidepressant-like effects: pharmacological and genetic evidences from the mouse forced swimming test. AB - Nociceptin/orphanin FQ (N/OFQ), the endogenous ligand of the NOP receptor, regulates several central functions such as pain transmission, learning and memory, fear and anxiety and feeding and locomotor activity. It has been recently reported that NOP receptor antagonists induce antidepressant-like effects in the mouse forced swimming test (FST), i.e. reduce immobility time. This assay was used in the present study for further investigating the involvement of the NOP receptor in depression states. In male Swiss mice, intracerebroventricular injection (i.c.v) of the novel NOP receptor antagonist, UFP-101 (1-10 nmol) dose dependently reduced the immobility time (control 192 +/- 14 s, UFP-101 91 +/- 15 s). The effect of 3 or 10 nmol UFP-101 was fully or partially reversed, respectively, by the coadministration of 1 nmol N/OFQ, which was inactive per se. NOP receptor knockout mice showed a reduced immobility time compared with their wild-type littermates (wild-type 215 +/- 10 s, knockout 143 +/- 12 s). Moreover, i.c.v. injected UFP-101 (10 nmol) significantly reduced immobility time in wild type mice but not in NOP receptor knockout animals. In conclusion, these results, obtained using a combined pharmacological and genetic approach, indicate that blockade of the N/OFQ-NOP receptor signalling in the brain produces antidepressant-like effects in the mouse FST. These findings support the NOP receptor as a candidate target for the development of innovative antidepressant drugs. PMID- 12752800 TI - Bioactivity of a peptide derived from acetylcholinesterase: electrophysiological characterization in guinea-pig hippocampus. AB - Acetylcholinesterase is well known to have noncholinergic functions. Only recently, however, has the salient part been identified of the molecule responsible for these nonclassical actions, a peptide of 14 amino acids towards the C-terminus of acetylcholinesterase. The aim of this study was to test the bioactivity of this 'acetylcholinesterase-peptide' using intracellular recordings in guinea-pig hippocampal slices. In the presence of tetrodotoxin, acetylcholinesterase-peptide alone affected neither the membrane potential nor the input resistance of CA1 neurons; however, a modulatory action was observed, as a concentration-dependent decrease of N-methyl-d-aspartic acid-induced depolarization. When calcium potentials were elicited by a depolarizing current pulse, application of acetylcholinesterase-peptide increased or reduced the degree of calcium spike firing in, respectively, the presence or absence of the N methyl-d-aspartic acid antagonist d(-)-2-amino-5-phosphonopentanoic acid. In contrast, a peptide derived from the equivalent region of butyrylcholinesterase, which also hydrolyses acetylcholine, had no effect. In conclusion, acetylcholinesterase-peptide has a selective bioactivity in the hippocampus; it could thus offer new ways of targeting mechanisms of calcium-induced neurotoxicity. PMID- 12752801 TI - Effect of various growth media upon survival during storage of freeze-dried Enterococcus faecalis and Enterococcus durans. AB - AIMS: The effects of three different growth media (MRS, M17 and Lee's) on survival during freeze-drying and subsequent storage of six strains of Enterococcus faecalis and two strains of E. durans were investigated. METHODS AND RESULTS: Distinct Enterococcus spp. strains were grown on M17, MRS and Lee's broth, freeze-dried and stored at 20 degrees C in air under darkness. At regular intervals throughout storage, freeze-dried samples were rehydrated and then plated on M17 agar. CONCLUSIONS: A higher survival rate during storage of dried E. durans was obtained when growth occurred in MRS. The same effect was not observed, however, for the majority of E. faecalis strains, which clearly survived better in the dried state when this organism had been grown in M17 or Lee's medium. SIGNIFICANCE AND IMPACT OF STUDY: The survival of the dried Enterococcus spp. tested during storage was shown to be strain-specific and dependent on the growth medium. PMID- 12752802 TI - Metal ion enhancement of fungal growth, gene expression and aflatoxin synthesis in Aspergillus flavus: RT-PCR characterization. AB - AIMS: To determine the effect of mineral ions (e.g., Zn2+, Cu2+, and Fe2+) on the enhancement of fungal growth, total RNA, aflatoxin pathway gene expression, and production of aflatoxin and its precursor O-methylsterigmatocystin (OMST). METHODS AND RESULTS: The influence of the metal ions, as a single or mixed treatments, was observed in submerged cultures of toxigenic Aspergillus flavus through changes in the fungal RNA or aflatoxin pathway gene (omtA) by reverse transcription-polymerase chain reaction (RT-PCR) levels, and also in fungal dry weight accumulation, aflatoxin and OMST production. CONCLUSIONS: The ion treatments induced changes of fungal total RNA, mRNA levels, associate fungal growth, biosynthesis of aflatoxin and OMST, and enhanced expression of RT-PCR. SIGNIFICANCE AND IMPACT OF STUDY: Demonstrates at the cellular and molecular level, the significant effects of metal ions on both fungal growth and production of aflatoxin. PMID- 12752803 TI - Modelling faecal indicator concentrations in large rural catchments using land use and topographic data. AB - AIMS: To characterize and model spatial variations in faecal-indicator organism concentrations in watercourses draining a large rural catchment during the bathing season. METHODS AND RESULTS: Presumptive coliform (PC), presumptive Eschericia coli (PE) and presumptive enterococci (PEnt) concentrations were determined under base- and high-flow conditions at 24 monitoring points in the Rheidol/Ystwyth catchment, Wales. Relationships with land use and topographic variables within their 'subcatchments' were investigated. Geometric mean (GM) concentrations typically increase more than 10-fold at high flow. The proportion of improved pasture within subcatchments explains 81.6, 73.0 and 85.8%, respectively, of the variance in high-flow GM PC, PE and PEnt concentrations. Distributed modelling suggests that organisms derived from more distant parts of subcatchments may be lost through die-off and sedimentation along watercourses at base flow, although not at high flow. Reductions in concentrations are also evident downstream of reservoir impoundments. CONCLUSIONS: Large rural catchments may contribute substantially to faecal indicator loadings in coastal waters. Indicator concentrations can be modelled successfully using land use and topographic data. SIGNIFICANCE AND IMPACT OF THE STUDY: Development of a modelling approach that provides valuable insight into faecal indicator sources and the transport and survival of these organisms within large catchments. PMID- 12752804 TI - Differential protein phosphorylation-dephosphorylation in response to carbon source in Ruminococcus flavefaciens FD-1. AB - AIMS: The aims of this study were to study the effect of cellobiose or cellulose as a carbon source on the differential protein phosphorylation-dephosphorylation of cytoplasmic and membrane-associated proteins from Ruminococcus flavefaciens FD 1. METHODS AND RESULTS: SDS-PAGE analysis was used to compare in vitro labelled proteins (32P-ATP) isolated from R. flavefaciens FD-1 grown on either cellobiose or cellulose as the carbon source. Distinctly different protein phosphorylation patterns were detected depending on carbon source and cell fraction. Analysis of the nature of the phosphorylated proteins indicates that phosphorylated proteins from cellobiose grown cultures are phosphorylated on serine residues, whereas phosphorylated proteins from cellulose grown cultures are phosphorylated on threonine residues. CONCLUSIONS: The results of this comparative analysis show a shift from serine phosphorylation of proteins to a threonine phosphorylation when R. flavefaciens FD-1 cells are grown on cellulose as opposed to cellobiose. There appears to be a role for these phosphorylation events in sensing the carbon source for growth and regulating co-ordinated metabolism in R. flavefaciens FD-1. SIGNIFICANCE AND IMPACT OF THE STUDY: We have demonstrated that there is a protein phosphorylation system in R. flavefaciens FD-1 that may be the primary sensing system for carbon source by R. flavefaciens FD-1 and the further regulation of gene expression related to cellulose degradation. PMID- 12752805 TI - Adhesion and aggregation ability of probiotic strain Lactobacillus acidophilus M92. AB - AIMS: To investigate aggregation and adhesiveness of Lactobacillus acidophilus M92 to porcine ileal epithelial cells in vitro, and the influence of cell surface proteins on autoaggregation and adhesiveness of this strain. METHODS AND RESULTS: Lactobacillus acidophilus M92 exhibits a strong autoaggregating phenotype and manifests a high degree of hydrophobicity determined by microbial adhesion to xylene. Aggregation and hydrophobicity were abolished upon exposure of the cells to pronase and pepsin. Sodium dodecyl sulphate polyacrylamide gel electrophoresis of cell surface proteins revealed the presence of potential surface layer (S layer) proteins, approximated at 45 kDa, in L. acidophilus M92. The relationship between autoaggregation and adhesiveness to intestinal tissue was investigated by observing the adhesiveness of L. acidophilus M92 to porcine ileal epithelial cells. Removal of the S-layer proteins by extraction with 5 mol l-1 LiCl reduced autoaggregation and in vitro adhesion of this strain. CONCLUSIONS: These results demonstrate that there is relationship between autoaggregation and adhesiveness ability of L. acidophilus M92, mediated by proteinaceous components on the cell surface. SIGNIFICANCE AND IMPACT OF THE STUDY: This investigation has shown that L. acidophilus M92 has the ability to establish in the human gastrointestinal tract, which is an important determinant in the choice of probiotic strains. PMID- 12752806 TI - Comparison of rapid methods for the extraction of bacterial DNA from colonic and caecal lumen contents of the pig. AB - AIMS: The increasing uses of DNA methodologies to study the micro flora of the pig gastrointestinal tract requires an efficient recovery of bacterial DNA from the intestinal sample. Thus, the objective of this study was to determine which DNA extraction methods are most effective for luminal samples from pigs. Several routinely used nucleic acid extraction procedures were compared based upon quantity and purity of extracted DNA. METHODS AND RESULTS: DNA was extracted from pig colonic and caecal lumen samples using 19 methods for bacterial DNA extraction. The quantity of total DNA recovered by each extraction method was determined and compared. Two methods using extraction with polyvinylpolypyrrolidone (PVPP) or phenol and two methods involving bead mill homogenization were found to provide the greatest quantity of extracted DNA for both colonic and caecal lumen. Extracted DNA from these four methods was further analysed for purity based upon the presence of PCR inhibitors, which was ascertained by determining the efficiency of amplification of a segment of the 16S rDNA. PCR amplification could be readily achieved with DNA extracted by each of these four methods, but efficiency of amplification tended to be higher with DNA from two of the methods (one extracted with PVPP and one with bead mill homogenization). CONCLUSIONS: Four extraction methods proved to be significantly superior in quantity of DNA extracted from luminal samples. Of these four, no strong inhibitors of PCR amplification were detected in any of the extracted DNA. However, the efficiency of amplification tended to be lower in DNA samples from two of the methods, suggesting the presence of low levels of PCR inhibitors. SIGNIFICANCE AND IMPACT OF THE STUDY: Results of this study provide a basis for choosing which DNA extraction procedures are most effective for use with samples of pig lumen. PMID- 12752807 TI - Comparison of enterococcal populations related to urban and hospital wastewater in various climatic and geographic European regions. AB - AIMS: Scarce knowledge about the distribution of enterococci species in wastewaters limits any statement on their reliability as faecal indicators or the implications of antibiotic resistance transmission by these organisms through the water cycle. Enterococci have been involved in nosocomial infections and the spreading of antibiotic resistance through the food chain. The species distribution of enterococci and the presence of resistant strains to vancomycin and erythromycin were analysed in more than 400 raw and treated urban wastewaters, surface waters receiving these treated wastewaters and hospital wastewaters from three European countries. METHODS AND RESULTS: A total of 9296 strains were isolated and biochemically phenotyped. The species identification was based on the comparison of biochemical profiles with those of more than 20000 enterococci isolates from an international study. The prevalence of enterococcal isolates resistant to erythromycin (ERE) and vancomycin (VRE) was also analysed. ERE strains were present in a high proportion in all the studied samples. VRE strains were also isolated in all studied countries despite the time elapsed since the use of antimicrobial glycopeptides in animal production was banned in the European Union. CONCLUSIONS: Enterococcus faecalis and Ent. faecium were the most abundant species in all the studied wastewaters. All the studied wastewaters demonstrated high diversity and similar population structure and composition. ERE and VRE isolates were detected in most of the wastewaters. SIGNIFICANCE AND IMPACT OF THE STUDY: Urban and hospital wastewaters are useful targets for the evaluation of the prevalence of ERE and VRE isolates in the environment. It appears that these bacteria could pass through wastewater treatment plants and be transferred to surface waters. PMID- 12752808 TI - PCR detection of seven virulence and toxin genes of Campylobacter jejuni and Campylobacter coli isolates from Danish pigs and cattle and cytolethal distending toxin production of the isolates. AB - AIMS: To study the prevalence of seven virulence and toxin genes, and cytolethal distending toxin (CDT) production of Campylobacter jejuni and C. coli isolates from Danish pigs and cattle. METHODS AND RESULTS: The presence of the cadF, ceuE, virB11, flaA, cdtA, cdtB, cdtC and the cdt gene cluster among 40 C. jejuni and C. coli isolates was detected by polymerase chain reaction. The CDT production of the isolates was determined on Vero, colon 205 and chicken embryo cells. The cadF, flaA, ceuE and cdtB genes were detected from 100% of the isolates. The cdtA and cdtC genes were found in 95.0 and 90.0% of the isolates, respectively. The cdt gene cluster was detected in 82.5% isolates. Only 7.5% of the isolates were positive for virB11. Ninety-five per cent of the isolates produced CDT in Vero and colon 205 cell assays, and 90% of the isolates produced CDT in chicken embryo cell assays. CONCLUSIONS: High prevalence of the cadF, ceuE, flaA and cdtB genes was found. Data of the prevalence of cdt genes was consistent with the CDT titres produced by the isolates. Campylobacter coli from pigs produced high CDT titres. SIGNIFICANCE AND IMPACT OF THE STUDY: The high prevalence of seven virulence and toxin genes demonstrated that these putative pathogenic determinants are widespread among Campylobacter isolates from pigs and cattle. Campylobacter coli isolates from pigs produced much higher CDT titres compared with C. coli isolates from other sources suggesting that C. coli may be particularly adapted to or associated with this species. PMID- 12752809 TI - Membrane damage to bacteria caused by single and combined biocides. AB - AIMS: To examine the effect on the leakage of low molecular weight cytoplasmic constituents from Staphylococcus aureus using phenolics singly and in combination, and to see if the observations could be modelled using a non-linear dose response. METHODS AND RESULTS: The rate of potassium, phosphate and adenosine triphosphate leakage was examined in the presence of chlorocresol and m cresol. Individually, leakage was observed only at long contact times or high concentrations. Combined at these ineffective concentrations, the cytoplasmic pool of all constituents studied was released within minutes. Both chlorocresol and m-cresol were shown to have non-linear dose responses. A rate model for the combinations, which takes account of these non-linear responses, accurately predicted the observations. CONCLUSIONS: Antimicrobials, which when used alone exhibit a non-linear dose response, will also give a non-linear dose response in combination. The simple linear-additive model ignores the concept of the dilution coefficient and will always describe the phenomenon of synergy for combinations where one or more of the components has a dilution coefficient greater than one. This has been borne out by examination of the purported prime lesion of chlorocresol and m-cresol, alone and in combination. SIGNIFICANCE AND IMPACT OF THE STUDY: Studies aimed at producing synergistic mixtures of antimicrobials, which ignore the non-linear additive effect, may waste valuable research effort looking for a physiological explanation for an apparent synergy, where none, in fact, exists. Patents granted on the basis of analyses using the linear-additive model for combinations of compounds with non-linear dose responses may no longer be supportable. PMID- 12752810 TI - Molecular fingerprinting evidence of the contribution of wildlife vectors in the maintenance of Salmonella Enteritidis infection in layer farms. AB - AIMS: To provide molecular fingerprinting evidence of the contribution of wildlife vectors in the on-farm epidemiology of Salmonella Enteritidis infections. METHODS AND RESULTS: Salmonella Enteritidis strains were isolated from wildlife and from farm environment samples collected in 10 egg layer farms. Isolates were typed using plasmid profiling, XbaI-pulsed field gel electrophoresis and PstI-SphI ribotyping. In all 10 farms we were able to identify the same S. Enteritidis clones in wildlife vectors and farm environment. On several occasions the same clones were found before and after cleansing and disinfecting the farm premises. Also in some instances the same clones were present in mice samples, egg contents and spent hens. CONCLUSIONS: Definitive molecular evidence for the involvement of several wildlife species (mice, rats, flies, litter beetles and foxes) in the maintenance of S. Enteritidis infection on farms has been presented. Failures in biosecurity seriously compromise the control of this pathogen on laying farms. SIGNIFICANCE AND IMPACT OF THE STUDY: This paper reports on the use of molecular tools for the study of the epidemiology of S. Enteritidis. It gives useful information to be considered in control programmes for this organism on poultry farms. PMID- 12752811 TI - Co-operative actions and degradation analysis of purified xylan-degrading enzymes from Thermomonospora fusca BD25 on oat-spelt xylan. AB - AIMS: To determine and quantify the products from the degradation of xylan by a range of purified xylan-degrading enzymes, endoxylanase, beta-xylosidase and alpha-l-arabinofuranosidase produced extracellularly by Thermomonospora fusca BD25. METHODS AND RESULTS: The amounts of reducing sugars released from oat-spelt xylan by the actions of endoxylanase, beta-xylosidase and alpha-l arabinofuranosidase were equal to 28.1, 4.6 and 7% hydrolysis (as xylose equivalents) of the substrate used, respectively. However, addition of beta xylosidase and alpha-l-arabinofuranosidase preparation to endoxylanase significantly enhanced (70 and 20% respectively) the action of endoxylanase on the substrate. The combination of purified endoxylanase, beta-xylosidase and alpha-l-arabinofuranosidase preparations produced a greater sugar yield (58.6% hydrolysis) and enhanced the total reducing sugar yield by around 50%. The main xylooligosaccharide products released using the action of endoxylanase alone on oat-spelt xylan were identified as xylobiose and xylopentose. alpha-l Arabinofuranosidase was able to release arabinose and xylobiose from oat-spelt xylan. In the presence of all three purified enzymes the hydrolysis products of oat-spelt xylan were mainly xylose, arabinose and substituted xylotetrose with lesser amount of substituted xylotriose. CONCLUSIONS: The addition of the beta xylosidase and alpha-l-arabinofuranosidase enzymes to purified xylanases more than doubled the degradation of xylan from 28 to 58% of the total substrate with xylose and arabinose being the major sugars produced. SIGNIFICANCE AND IMPACT OF THE STUDY: The results highlight the role of xylan de-branching enzymes in the degradation of xylan and suggest that the use of enzyme cocktails may significantly improve the hydrolysis of xylan in industrial processes. PMID- 12752812 TI - Decontamination of pork carcasses during scalding and the prevention of Salmonella cross-contamination. AB - AIMS: The objective of this study was to establish critical temperature limits to prevent cross-contamination of pork carcasses during scalding. METHODS AND RESULTS: Mixtures of antibiotic-resistant mutants of Salmonella species were heat treated at 50, 55 and 60 degrees C in samples of commercial scald tank water. Surviving cell numbers were estimated by plating treated suspensions on (i). tryptone soya agar (TSA) and (ii). on TSA, overlaid with brilliant green agar plus nalidixic acid and streptomycin sulphate and used to estimate D-values for the treated mixed cell suspensions. CONCLUSIONS: A time-temperature combination of 1.4 min at 60 degrees C is required to achieve a 1 log reduction in Salmonella in scald tank water. The predicted equivalent at 65 degrees C is 0.18 min. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides data and a model to enable pork processors to identify and apply processing parameters to limit the risks of transfer of Salmonella between pig carcasses during commercial scalding operations. PMID- 12752813 TI - Atrazine catabolism by a combined bacterial association (KRA30) under carbon- and nitrogen-limitations in a retentostat. AB - AIMS: Nutrient-limited atrazine catabolism study in continuous cultures with biomass retention to mimic in situ environmental conditions and thus gain insight of the efficacy of biosupplementation/biostimulation to eliminate reduced herbicide bioavailability. METHODS AND RESULTS: Carbon- and nitrogen-limited retentostat (1 and 5 l) cultivation of a combined atrazine (100 mg l-1) catabolizing association KRA30 was made. As a nitrogen source, through citrate supplementation, increased herbicide catabolism resulted and was complete in the absence of NH4-N. Co-metabolism of the molecule in the presence of succinate was identified. Population characterization by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) indicated component species numerical dominance shifts in response to changes in nutrient limitation, mineral salts composition and biofilm formation, although the total species complement and catabolic potential were retained. CONCLUSIONS: Biomass and catabolic capacity maintenance, through cost-effective biosupplementation/biostimulation, should promote atrazine bioavailability and so ensure successful amelioration. SIGNIFICANCE AND IMPACT OF THE STUDY: All planning, implementation and monitoring of bioremediation programmes should be underpinned by a combination of molecular and (continuous) culture-based methods. PMID- 12752814 TI - Correlation between bacterial haemoglobin gene (vgb) and aeration: their effect on the growth and alpha-amylase activity in transformed Enterobacter aerogenes. AB - AIMS: To evaluate the effects of bacterial haemoglobin on bacterial growth and alpha-amylase formation under different aeration conditions. METHODS AND RESULTS: Enterobacter aerogenes was transformed with the gene encoding Vitreoscilla (bacterial) haemoglobin, vgb. The growth kinetics and ability to synthesize alpha amylase enzyme were investigated in this transformed Enterobacter strain as well as in two other Enterobacter control strains that do not harbour the vgb gene. Such comparison was made under variable aeration conditions, using the agitation rate as a measure of aeration. The expression of bacterial haemoglobin-supported cell growth determined as O.D.600 and cell viability in addition to the alpha amylase production. These positive effects of bacterial haemoglobin were observed under both low and high aerations, but at different extents. CONCLUSIONS: In addition to improving cell growth under low aeration, the bacterial haemoglobin is able to promote bacterial cell tolerance during exposure to high oxygen tension. SIGNIFICANCE AND IMPACT OF THE STUDY: The expression of bacterial haemoglobin is advantageous in reducing the burden of certain toxic conditions such as high oxygen levels. It may have the same impact on some environmental toxic substances. This, haemoglobin biotechnology can be extended to induce enzymes of pollutants degradation or production of some useful industrial substances. PMID- 12752815 TI - Starch in plasterboard sustains Streptomyces californicus growth and bioactivity of spores. AB - AIMS: The effects of plasterboard composition on Streptomyces californicus growth and bioactivity of spores were studied. METHODS AND RESULTS: Streptomyces californicus was grown on 13 modified plasterboards under saturated humidity conditions. The total content of fatty acid methyl esters was used for quantifying S. californicus biomass, while the spore-induced cytotoxicity and production of nitric oxide (NO), tumour necrosis factor-alpha, and interleukine-6 (IL-6) in mouse macrophages was used to assess the bioactivity of spores. Removal of starch completely from the plasterboard or only from the core reduced significantly the biomass production and the biological activity of spores in comparison with reference board. The biocide added into the core or on the liner decreased the growth markedly and inhibited the sporulation totally. The biomass production correlated positively with the spore number, cytotoxicity, and production of NO and IL-6. CONCLUSIONS: Streptomyces californicus grew under nutrient limitation on all studied plasterboards. The starch is the major factor enabling S. californicus to grow and to produce biologically active metabolites on plasterboard. SIGNIFICANCE AND IMPACT OF THE STUDY: The composition of building material has an impact on microbial growth and bioactivity of spores which may be involved in complex mechanisms leading to respiratory symptoms in the occupants in moisture damaged buildings. PMID- 12752816 TI - The survival of silage inoculant lactic acid bacteria in rumen fluid. AB - AIMS: To determine whether lactic acid bacteria (LAB) used in inoculants for silage can survive in rumen fluid (RF), and to identify those that survive best. METHODS AND RESULTS: Twelve commercial silage inoculants were added at 107 CFU ml 1 to strained RF (SRF) taken from dairy cows, with and without 5 g l-1 glucose and incubated in vitro at 39 degrees C. Changes in pH, LAB numbers and fermentation products were monitored for 72 h. In the inoculated RF with glucose, the pH decreased and numbers of LAB increased. The inoculants varied with regard to their effect on pH change and growth. In the SRF, both with and without glucose, the pH values of the inoculated samples were generally higher than those of the uninoculated controls throughout most of the incubation period. This may suggest a positive effect on the rumen environment. CONCLUSIONS: LAB used in silage inoculants can survive in RF in vitro. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first step in studying the probiotic potential of silage LAB inoculants for dairy cattle. The survival of these LAB in RF may enable them to interact with rumen microorganisms and to affect rumen functionality. PMID- 12752817 TI - Factors associated with the adherence and biofilm formation by Aeromonas caviae on glass surfaces. AB - AIMS: Knowledge on factors of Aeromonas caviae promoting the formation of biofilms on surfaces. METHODS AND RESULTS: In nutrient broth under agitation, A. caviae LMG 13455 was able to form biofilms on the surfaces of glass flasks, but such biofilm formation was inconsistent. A derivative of this strain, called M12, promoted the rapid formation of reliable and strongly adherent biofilms with about half of the cells being adhered. In contrast with its parent, M12 was hydrophobic, displayed auto-aggregation in liquid medium, synthesized very little polysaccharides and was defective in swimming and swarming motilities, together with the appearance of a characteristic phenotype on motility soft agar. Further analyses demonstrated that most of these properties were related to a hyperpiliation of the cells through the presence of type IV pili, and suggested that a mechanism of genetic variation, by altering the nature of motility appendages, allows the variant bacteria to attach on inert surfaces. CONCLUSIONS: M12 is a stabilized variant of the parental strain, promoting strongly adherent biofilms through the type IV pili. SIGNIFICANCE AND IMPACT OF THE STUDY: The wild type strain A. caviae LMG 13455 include subpopulations that are likely implicated in its adaptation to different environmental changes. PMID- 12752818 TI - Taxonomic positioning of two biological control agents for plant diseases as Lysobacter enzymogenes based on phylogenetic analysis of 16S rDNA, fatty acid composition and phenotypic characteristics. AB - AIMS: To taxonomically position two bacterial strains conferring biological control activity towards plant diseases. METHODS AND RESULTS: Key phenotypic characteristics, including gliding motility and a high percentage of G + C content, indicated biocontrol strains N4-7 and C3 were essentially identical to those described for Lysobacter enzymogenes. Cellular fatty acid analysis confirmed a close relatedness of strains N4-7 and C3 to L. enzymogenes and a more distant relatedness to L. antibioticus. The 16S rDNA phylogenetic analysis revealed a distinct Lysobacter clade that included both strains within the gamma proteobacteria. CONCLUSIONS: The combined taxonomic methods provide clear evidence that N4-7 and C3 should be grouped as strains of L. enzymogenes and not Stenotrophomonas maltophilia or a novel taxon. Phylogenetic analysis of 16S rDNA formed a Lysobacter clade that included several other environmentally diverse bacterial strains obtained from databases and confirmed relatedness of strains N4 7 and C3 to L. enzymogenes. SIGNIFICANCE AND IMPACT OF THE STUDY: Inclusion of N4 7 and C3 as strains of L. enzymogenes is among the first description of members of this genus as biocontrol agents of plant diseases. These results suggest that members of the Lysobacter group might provide a new source as plant-associated microbes that display biocontrol activity. PMID- 12752819 TI - Influence of tetracycline exposure on tetracycline resistance and the carriage of tetracycline resistance genes within commensal Escherichia coli populations. AB - AIMS: To assess the influence of incremental tetracycline exposure on the genetic basis of tetracycline resistance within faecal Escherichia coli. METHODS AND RESULTS: Through the adoption of a novel combination of multiple breakpoint selection, phenotypic characterization and the application of a polymerase chain reaction based gene identification system it proved possible to monitor the influence of antibiotic exposure on resistance gene possession. Using tetracycline as a case study a clear hierarchy was revealed between tet genes, strongly influenced by host antimicrobial exposure history. CONCLUSIONS: The antimicrobial exposure regime under which an animal is produced affects both the identity and magnitude of resistance gene possession of a selected bacterial population within its enteric microflora. Among the ramifications associated with such resistance gene selection is the degree of resistance conferred and the carriage of linked resistance determinants. This selection is applied by exposure to antibiotic concentrations well below recognized minimum inhibitory tetracycline concentration breakpoints widely adopted to characterize bacterial 'susceptibility'. SIGNIFICANCE AND IMPACT OF THE STUDY: This study confirms the ability of minimal antibiotic exposure to select for the continued persistence of resistance genes within the enteric microflora. It is clearly demonstrated that different antimicrobial regimes select for different resistance genes, the implications of which are discussed. PMID- 12752820 TI - Phenotypic and genotypic characterization of competitive exclusion products for use in poultry. AB - AIMS: Phenotypic and genotypic bacteria identification methods were compared for their efficacy in determining the composition of competitive exclusion (CE) products. METHODS AND RESULTS: Phenotypic methods used for bacterial identification were fatty acid methyl ester profiles, biochemical assays and carbohydrate utilization profiles. Genotypic methods were MicroSeq16S rRNA sequence analysis and BLAST searches of the GenBank sequence database. Agreement between phenotypic and genotypic methods for identification of bacteria isolated from the Preempt CE product was 20%. A defined test mixture of bacteria was identified to the species level 100% by BLAST analysis, 64% by MicroSeq and 36% by phenotypic techniques. CONCLUSIONS: The wide range of facultative and obligate anaerobic bacteria present in a CE product are more accurately identified with 16S rRNA sequence analyses than with phenotypic identification techniques. SIGNIFICANCE AND IMPACT OF THE STUDY: These results will provide guidelines for manufacturers of CE products to submit more reliable product information for market approval by regulatory agencies. PMID- 12752821 TI - Genetic relationship in the 'Bacillus cereus group' by rep-PCR fingerprinting and sequencing of a Bacillus anthracis-specific rep-PCR fragment. AB - AIMS: To evaluate the genetic relationship in the Bacillus cereus group by rep PCR fingerprinting. METHODS AND RESULTS: A collection of 112 strains of the six species of the B. cereus group was analysed by rep-PCR fingerprinting using the BOX-A1R primer. A relative genetic distinctness was found among the species. Cluster analysis of the rep-PCR patterns showed clusters of B. thuringiensis strains quite separate from those of B. cereus strains. The B. anthracis strains represented an independent lineage in a B. cereus cluster. The B. mycoides, B. pseudomycoides and B. weihenstephanensis strains were clustered into three groups at some distance from the other species. Comparison of sequences of AC-390, a typical B. anthracis rep-PCR fragment, from 27 strains of B. anthracis, B. cereus, B. thuringiensis and B. weihenstephanensis, representative of different clusters identified by rep-PCR fingerprinting, confirmed that B. anthracis diverges from its related species. CONCLUSIONS: The genetic relationship deduced from the rep-PCR patterns indicates a relatively clear separation of the six species, suggesting that they can indeed be considered as separate units. SIGNIFICANCE AND IMPACT OF THE STUDY: rep-PCR fingerprinting can make a contribution in the clarification of the genetic relationships between the species of the B. cereus group. PMID- 12752822 TI - Purification and characterization of a membrane glycoprotein from the fish pathogen Flavobacterium psychrophilum. AB - AIMS: The cell envelope of the fish pathogen Flavobacterium psychrophilum contains more than 50 polypeptides resolved by sodium dodecyl sulphate polyacrylaminde gel electrophoresis analysis including a major component named P60. Here, we have developed a simple and efficient procedure for the purification of P60 and therefore permitting its biochemical characterization. METHODS AND RESULTS: Membrane proteins were selectively extracted from isolated cell envelopes with the mild non-ionic detergent Triton X-100. About 10 polypeptides were identified from the detergent fraction, including P60. The P60 enriched fraction was thereafter subjected to an anion exchange chromatographic step in the presence of Triton X-100. The molecule was purified at the milligram level (yield, about 75%; purification factor, 6.2). Analyses performed by charge shift electrophoresis, Triton X-114 phase separation and by detection of sugar modified components showed that P60 is a true amphiphilic membrane-associated glycoprotein. CONCLUSIONS: The method described in this paper provides pure and non-denaturated P60 and should prove to be easily scaled-up. As sugar-modified protein, P60 should be included in the growing list of glycosylated prokaryotic proteins. SIGNIFICANCE AND IMPACT OF THE STUDY: It offers the possibility of obtaining P60 in amounts allowing the testing of the potential of P60 as a candidate for anti-flavobacteria subunit vaccines, as P60 is one of the major antigens. PMID- 12752823 TI - Influence of dietary beetpulp on the plasma level of triacylglycerols in horses. AB - In a cross-over study with six adult horses, the effect of isoenergetic replacement of dietary glucose by beetpulp on the concentration of plasma triacylglycerols was studied. The test ration contained 25% beetpulp in the dietary dry matter. The feeding of beet pulp lowered plasma triacylglycerol concentrations (p = 0.058) in the fasting state and raised the activity of heparin-released lipoprotein lipase (LPL) (p = 0.059) and the concentration of HDL2 cholesterol (p = 0.058). In the fasting state, the plasma concentrations of free fatty acids (p = 0.073) and those of beta-hydroxybutyrate (p = 0.043) were increased after feeding beetpulp. At 3 h after feed intake, consumption of beetpulp instead of glucose had produced higher plasma concentrations of both glucose (p = 0.014) and insulin (p = 0.040). It is suggested that beetpulp ingestion activates LPL followed by a drop in plasma triacylglycerols. PMID- 12752824 TI - Production and clearance of plasma triacylglycerols in ponies fed diets containing either medium-chain triacylglycerols or soya bean oil. AB - The hypothesis was tested that feeding ponies a diet containing medium-chain triacylglcyerols (MCT) instead of soya bean oil causes an increase in the production of plasma triacylglycerols, which, under steady-state conditions, is associated with an increased clearance of triacylglycerols. Six ponies were fed rations containing either MCT or an isoenergetic amount of soya bean oil according to a cross-over design. The concentration of MCT in the total dietary dry matter was about 13%. When the ponies were fed the diets for 3 weeks, plasma triacylglycerol concentrations were 0.42 +/- 0.09 and 0.17 +/- 0.03 mmol/l (mean +/- SE, n = 6; p < 0.05) for the MCT and soya bean-oil treatment, respectively. Plasma triacylglycerol production was assessed using the Triton method and clearance with the use of Intralipid(R) infusion. Plasma triacylglycerol production was 2.91 +/- 0.88 and 0.50 +/- 0.14 micromol/l.min (means +/- SE, n = 4; p < 0.05) for the diets containing MCT and soya bean oil, respectively. It is suggested that the calculated rates of triacylglycerol production are underestimated, the deviation being greatest when the ponies were fed the ration of soya bean oil. Triacylglycerol clearance rates were calculated on the basis of group mean values for both the fractional clearance rate and the baseline levels of plasma triacylglycerols; the values were 4.28 and 3.52 micromol/l.min for MCT and soya bean oil feeding, respectively. The mean, absolute clearance rates as based on those found in individual ponies did not show an increase when the diet with MCT was fed. Nevertheless, it is concluded that the data obtained support our hypothesis. PMID- 12752825 TI - Estimation of the proportion of body fat in mice from the proportion of body water. AB - We compared the proportion of body fat in mice as measured by chemical analysis with that estimated from the proportion of body water. First, we measured the proportion of fat by chemical analysis in 78 mice that had a proportion of body fat in the range from approximately 5 to 20%. Then, we constructed a regression line that described the relationship between the proportion of body water and the proportion of body fat by using data from several other published studies in mice (% body fat = -1.20 x % body water + 88.07, r = 0.9597, sy.x = 2.75, p < 0.001). With this regression line, we estimated the proportion of body fat from the proportion of body water that was measured by drying the carcasses at 60 degrees C for 3 days. Body fat data obtained from this regression line were similar to those obtained by chemical analysis. Thus, these results suggest that reliable values for the proportion of body fat can be derived from the proportion of body water and this method provides a tool to rapidly measure the proportion of body fat in mice. PMID- 12752826 TI - Assessment of the minimum protein requirement of adult ponies. AB - The aim of the present study was to determine the minimum protein requirement in adult ponies. Literature data from nitrogen-balance trials in adult ponies were used. In addition, nitrogen balance was measured in three mature ponies that were fed a low- and a high-protein diet according to a cross-over design. On the low protein ration, which provided 1.1 g digestible crude protein (DCP)/kg(0.75) day, plasma albumin and total protein concentrations were non-significantly decreased by about 6% and group-mean nitrogen balance was negative. Using data from the present experiment and from six other studies, the sum of endogenous faecal and urinary nitrogen losses was calculated to be 166 +/- 37 mg/kg(0.75) day (mean +/- SE, n = 13). Subsequently, a minimum protein requirement of 1.7 g DCP/kg(0.75) day was calculated. The value for minimum protein requirement has low precision, which should be taken into account when setting the protein allowance of adult ponies. PMID- 12752827 TI - Amino acid, mineral and vitamin levels in hydrous faeces obtained from coprophagy prevented rats. AB - Amino acid, mineral and vitamin levels in hydrous faeces obtained from coprophagy prevented rats were examined. Two experiments were conducted with rats of different age, 4 weeks and 10 weeks. Six rats each were fed under coprophagy prevented and coprophagy-allowed (conventional feeding) conditions, respectively. In the first experiment using 4-week-old rats, amino acid values were determined for hydrous faeces and usual faeces obtained in the above-mentioned conditions. In the second experiment using 10-week-old rats, mineral values were determined for both types of faeces. Vitamin values were determined in both experiments. In a comparison of amino acid values and their patterns, specific kinds of amino acids in hydrous faeces were not recognized as growth factors. Sodium and potassium levels in hydrous faeces were significantly higher than those for usual faeces, but those levels were 1/(18-20) of those derived from feed. Although cyanocobalamin and folic acid levels in hydrous faeces were two to three times higher than those for usual faeces, folic acid supplements from coprophagy were 1/8 of those derived from feed. Cyanocobalamin supplements from hydrous faeces were five times larger than those from feed. Therefore, it appears that cyanocobalamin was supplied by the ingestion of hydrous faeces and that this cyanocobalamin supports conventional growth in rats. PMID- 12752828 TI - Nutrition studies on protein and energy in domestic cats. AB - The effects of three diets varying in their protein, fat and carbohydrate contents and of physical activity on energy and protein metabolism were investigated in eight domestic cats. The diets were based on canned food with addition either of minced cattle heart, melted swine fat or polenta. The activity induced heat production was measured in a respiration chamber using the Doppler effect of radio-waves reflected from moving bodies. In the group receiving the high-protein diet, urinary nitrogen concentration was higher which was reflected in higher urinary loss of energy without affecting the amount of energy retained. Nitrogen utilization was not significantly influenced by nitrogen intake. Mean daily total heat production ranged between 163 and 187 kJ/kg BW/day or 215 and 270 kJ/kg BW(0.75)/day. The daily crude protein requirement of adult cats was determined to be 2.7 g/kg BW/day. The daily requirement of metabolizable energy for maintenance was estimated to be 153 kJ/kg BW/day or 226 kJ/kg BW(0.75)/day. The activity-induced heat production was 25.1 kJ/kg BW/day which corresponded to 13.5% of the total daily heat production. The daily requirement for metabolizable energy was thus reduced by 20.9 kJ/kg BW/day or 13.7%. The activity pattern of the cats was not characterized by a circadian rhythm. PMID- 12752829 TI - Effect of dietary rare earth elements on growth performance and blood parameters of rats. AB - Rare earth elements (REE) have been shown to influence growth performance in animal production, especially in pigs. In the present study, the effect of oral administration of rare earth elements on growing rats was investigated. Pure LaCl3 or an REE mixture containing 38% of LaCl3, 52% of CeCl3, 3% of PrCl3 and 7% of chlorides of other REE were used at two different concentrations as supplements to the diets. Fifty male Wistar rats at 4 weeks of age were allotted to five experimental groups: a control group; a La-low group and a La-high group with 75 and 150 mg/kg LaCl3.6H2O, respectively; a REE-low and an REE-high group with 75 and 150 mg/kg REE mixture, respectively. The animals were housed in individual pens. Feed and water were provided ad libitum. After 18 days the oral supplementation of LaCl3.6H2O or of the REE mixture improved daily body weight gain (BWG) by up to 5 or 9% (p > 0.05), respectively. LaCl3.6H2O as well as the REE mixture had positive effects (p < 0.05) on feed conversion ratio (FCR) with a decreased ratio by up to 8 and 11%, respectively. Supplementation of REE also had clear effects on blood serum parameters. The activities of alkaline phosphatase (AP) and alanine amino transferase (ALT) increased significantly (p < 0.05). At the same time, blood glucose level decreased and blood creatine level increased significantly (p < 0.05). There was no significant difference in cholesterol, total protein, albumin and urea nitrogen among the groups. There was no significant difference in triglyceride level between the control and those REE groups, however, a significantly lower (p < 0.01) triglyceride level was found in the 150 mg/kg REE mixture group compared with that in 75 mg/kg REE mixture group and the 150 mg/kg LaCl3.6H2O group. The results suggest that oral supplementation of REE improves growth performance in rats as in pigs. In this respect, concentration and type of REE supplemented to the diets are two important factors herein. PMID- 12752830 TI - Plasma and whole blood taurine in normal dogs of varying size fed commercially prepared food. AB - The objective of the present study was to examine the effect of signalment, body size and diet on plasma taurine and whole blood taurine concentrations. A total of 131 normal dogs consuming commercially prepared dog food had blood drawn 3-5 h post-prandially to be analysed for plasma amino acids and whole blood taurine. Body weight and morphometric measurements of each dog were taken. Plasma and whole blood taurine concentrations were 77 +/- 2.1 nmol/ml (mean +/- SEM) and 266 +/- 5.1 nmol/ml (mean +/- SEM), respectively. No effect of age, sex, body weight, body size, or diet was seen on plasma and whole blood taurine concentrations. Mean whole blood taurine concentrations were lower in dogs fed diets containing whole grain rice, rice bran or barley. The lowest whole blood concentrations were seen in dogs fed lamb or lamb meal and rice diets. Plasma methionine and cysteine concentrations were lower in dogs fed diets with animal meals or turkey, and whole grain rice, rice bran or barley. Fifteen of 131 dogs had plasma taurine concentrations lower than, or equal, to the previously reported lowest mean food deprived plasma taurine concentration in normal dogs of 49 +/- 5 nmol/ml (mean +/ SEM) (Elliott et al., 2000). These findings support the theory that taurine deficiency in dogs may be related to the consumption of certain dietary ingredients. Scientific and clinical evidence supports the hypothesis that dilated cardiomyopathy is associated with low blood taurine concentration in dogs; therefore, further work is indicated to determine the mechanism by which diet can affect taurine status in dogs. PMID- 12752832 TI - Delirium in the intensive care unit: occurrence and clinical course in older patients. AB - OBJECTIVES: To describe the occurrence of delirium in a cohort of older medical intensive care unit (ICU) patients and its short-term duration in the hospital and to determine the association between preexisting dementia and the occurrence of delirium. DESIGN: Prospective cohort study. SETTING: Fourteen-bed medical ICU of an 800-bed university teaching hospital. PARTICIPANTS: One hundred eighteen consecutive patients aged 65 and older admitted to the ICU. MEASUREMENTS: Baseline characteristics were obtained through surrogate interviews and medical chart review. Dementia was determined using two validated surrogate-rated instruments. Delirium was assessed daily in the ICU using the Confusion Assessment Method (CAM) for the ICU (CAM-ICU). After discharge from the ICU, patients were followed for up to 7 days using the CAM. RESULTS: Delirium was present in 37 of 118 (31%) patients on admission. Only 45 patients had a normal mental status on admission, of whom 14 (31%) became delirious during their hospital stay. In the post-ICU period, delirium occurred in 40% of patients. Almost half of patients with delirium in the ICU had persistent delirium in the post-ICU period. Overall, 83 of 118 (70%) had delirium during hospitalization. Stupor or coma occurred in 44% of the patients overall, and 89% of survivors of stupor/coma progressed to delirium. Patients with dementia were 40% more likely to be delirious (relative risk = 1.4, 95% confidence interval = 1.1-1.7), even after controlling for comorbidity, baseline functional status, severity of illness, and invasive procedures. CONCLUSION: Delirium is a frequent complication in older ICU patients and often persists beyond their ICU stay. Delirium in older ICU persons is a dynamic and complex process. Dementia is an important predisposing risk factor for the development of delirium in this population during and after the ICU stay. PMID- 12752833 TI - Efficacy of percutaneous electrical nerve stimulation for the treatment of chronic low back pain in older adults. AB - OBJECTIVES: To determine the efficacy of a complementary analgesic modality, percutaneous electrical nerve stimulation (PENS), for the treatment of chronic low back pain (CLBP) in community-dwelling older adults. DESIGN: Randomized, controlled clinical trial. SETTING: University of Pittsburgh Pain Evaluation and Treatment Institute. PARTICIPANTS: Thirty-four English speaking, community dwelling adults aged 65 and older with CLBP of at least moderate intensity experienced every day or almost every day. INTERVENTION: Subjects were randomized to receive twice-weekly PENS and physical therapy (PT) or sham PENS and physical therapy for 6 weeks. MEASUREMENTS: At baseline, immediately after the 6-week intervention period, and 3 months later, the primary outcome measures pain intensity and pain-related disability were assessed. The secondary outcome measures physical performance (timed chair rise, functional reach, gait speed, static and isoinertial lifting), psychosocial factors (mood, sleep, and life control), and cognitive function (measures of attention, concentration, and mental flexibility) were also collected. RESULTS: Subjects randomized to PENS plus PT displayed significant reductions in pain intensity measures from pre- to posttreatment (P <.001), but the sham PENS plus PT group did not (P =.94). These pain reduction effects were maintained at 3-month follow-up. Similarly, significant reductions in pain-related disability were observed at posttreatment (P =.002) for the PENS plus PT group and were maintained at follow-up, but the sham PENS plus PT group did not show reductions in pain-related disability (P =.81). Of the secondary outcome measures, psychosocial function, timed chair rise, and isoinertial lifting endurance also improved significantly at posttreatment for the PENS plus PT group, and their improvement was sustained at 3-month follow-up, but the sham PENS plus PT did not display significant changes on these measures after treatment. CONCLUSION: This preliminary study suggests that PENS may be a promising treatment modality for community-dwelling older adults with CLBP, as demonstrated by reduction in pain intensity and self reported disability, and improvement in mood, life control, and physical performance. Larger studies with longer duration of follow-up are needed to validate these findings and support the use of PENS in clinical practice. PMID- 12752834 TI - Using self-reported data to predict expenditures for the health care of older people. AB - OBJECTIVES: To create and test a method for using self-reported data to predict future expenditures for the health care of older people. DESIGN: A two-stage regression model of the relationship between self-reported data and Medicare expenditures during the following year was constructed from a randomly selected (derivation) half of a cohort of fee-for-service Medicare beneficiaries. For the other (validation) half of the cohort, two sets of predictions of 12-month Medicare expenditures were generated, one using the new two-stage model and the other using the principal inpatient diagnostic cost group (PIP-DCG) method now used to risk-adjust capitation payments to Medicare + Choice health plans. Both sets of predictions were compared with Medicare's actual 12-month expenditures for the validation cohort. SETTING: Ramsey County, Minnesota. PARTICIPANTS: Community-dwelling Medicare beneficiaries aged 70 and older (N = 13,682) who responded to a mailed survey. MEASUREMENTS: Predicted-to-observed ratio (PTOR) of Medicare expenditures. RESULTS: For the validation cohort, Medicare's actual 12 month expenditures totaled $26.5 million. The two-stage model predicted Medicare expenditures of $26.4 million (PTOR = 1.00); the PIP-DCG method predicted $31.2 million (PTOR = 1.18). Within subpopulations of healthy and ill beneficiaries, the two-stage model's predictions remained considerably more accurate than the PIP-DCG predictions. CONCLUSION: Self-reported data may predict future Medicare expenditures more accurately than administrative data about beneficiaries' demographic characteristics, and previous hospitalizations. PMID- 12752835 TI - Identification of risk for high hospital use: cost comparisons of four strategies and performance across subgroups. AB - OBJECTIVES: To determine the relative costs of four risk-identification strategies and compare their performance in predicting hospital use by different subgroups of older persons based on age, sex, and prior hospital use. DESIGN: Prospective validation study and cost-comparison analysis. SETTING: Community based. PARTICIPANTS: Five thousand one hundred thirty-eight participants of the sixth wave of three sites of the Established Populations for Epidemiologic Studies of the Elderly. MEASUREMENTS: Four strategies (prior hospitalization data only, a 10-item self-report screen alone, self-report combined with two laboratory tests, and sequential self-report plus as-needed use of laboratory tests when the self-report screen is inconclusive) and 3-year Medicare Part A hospital cost data. RESULTS: Assuming that interventions based on screening would yield a total benefit of $1,000 per true-positive case and a cost of $400 for each false-positive case, the sequential strategy was slightly less expensive than the self-report only strategy; both were considerably less expensive than the combined or hospitalization-only strategies. Accuracy as measured by the area under the receiver operating characteristic curve for the sequential strategy was comparable for all subgroups (between 0.62 and 0.70) but was least accurate for those who had high prior use and for those aged 85 and older. CONCLUSION: A sequential screening strategy that administers laboratory tests selectively is slightly less expensive than one that uses only self-report items. This strategy is also accurate in both sexes, in those with various degrees of prior use, and in the oldest old. PMID- 12752836 TI - Unsteadiness reported by older hospitalized patients predicts functional decline. AB - OBJECTIVES: To determine whether a simple question about steadiness at admission predicts in-hospital functional decline and whether unsteadiness at admission predicts failure of in-hospital functional recovery of patients who have declined immediately before hospitalization. DESIGN: Prospective cohort study. SETTING: One university hospital and one community teaching hospital. PARTICIPANTS: One thousand five hundred fifty-seven hospitalized medical patients aged 70 and older. MEASUREMENTS: On admission, patients reported their steadiness with walking and whether they could perform independently each of five basic activities of daily living (ADLs) at admission and 2 weeks before admission (baseline). For the primary analysis, the outcome was decline in ADL function between admission and discharge. For the secondary analysis, the outcome was in hospital recovery to baseline ADL function in patients who experienced ADL decline in the 2 weeks before admission. RESULTS: In the primary cohort (n = 1,557), 25% of patients were very unsteady at admission; 22% of very unsteady patients declined during hospitalization, compared with 17%, 18%, and 10% for slightly unsteady, slightly steady, and very steady patients, respectively (P for trend =.001). After adjusting for age; medical comorbidities; Acute Physiology, Age, and Chronic Health Evaluation II score; and admission ADL, unsteadiness remained significantly associated with ADL decline (odds for decline for very unsteady compared with very steady = 2.6, 95% confidence interval = 1.5-4.5). In the secondary analysis, predicting ADL recovery in patients who declined before hospitalization (n = 563), 46% of patients were very unsteady at admission. In this cohort, 44% of very unsteady patients failed to recover, compared with 35%, 36%, and 33% for each successively higher level of steadiness, respectively (P for trend = 0.06). After multivariate adjustment, greater unsteadiness independently predicted failure of recovery (P for trend = 0.02). CONCLUSION: A simple question about steadiness identified patients at increased risk for in hospital ADL decline and, in patients who lost ADL function immediately before admission, failure to recover. PMID- 12752837 TI - Fall and injury prevention in residential care--effects in residents with higher and lower levels of cognition. AB - OBJECTIVES: To evaluate the effectiveness of a multifactorial fall and injury prevention program in older people with higher and lower levels of cognition. DESIGN: A preplanned subgroup comparison of the effectiveness of a cluster randomized, nonblinded, usual-care, controlled trial. SETTING: Nine residential facilities in Umea, Sweden. PARTICIPANTS: All consenting residents living in the facilities, aged 65 and older, who could be assessed using the Mini-Mental State Examination (MMSE; n = 378). An MMSE score of 19 was used to divide the sample into one group with lower and one with higher level of cognition. The lower MMSE group was older (mean +/- standard deviation = 83.9 +/- 5.8 vs 82.2 +/- 7.5) and more functionally impaired (Barthel Index, median (interquartile range) 11 (6-15) vs 17 (13-18)) and had a higher risk of falling (64% vs 36%) than the higher MMSE group. INTERVENTION: A multifactorial fall prevention program comprising staff education, environmental adjustment, exercise, drug review, aids, hip protectors, and postfall problem-solving conferences. MEASUREMENTS: The number of falls, time to first fall, and number of injuries were evaluated and compared by study group (intervention vs control) and by MMSE group. RESULTS: A significant intervention effect on falls appeared in the higher MMSE group but not in the lower MMSE group (adjusted incidence rates ratio of falls P =.016 and P =.121 and adjusted hazard ratio P <.001 and P =.420, respectively). In the lower MMSE group, 10 femoral fractures were found, all of which occurred in the control group (P =.006). CONCLUSION: The higher MMSE group experienced fewer falls after this multifactorial intervention program, whereas the lower MMSE group did not respond as well to the intervention, but femoral fractures were reduced in the lower MMSE group. PMID- 12752838 TI - Handgrip strength and cause-specific and total mortality in older disabled women: exploring the mechanism. AB - OBJECTIVES: To examine the association between muscle strength and total and cause-specific mortality and the plausible contributing factors to this association, such as presence of diseases commonly underlying mortality, inflammation, nutritional deficiency, physical inactivity, smoking, and depression. DESIGN: Prospective population-based cohort study with mortality surveillance over 5 years. SETTING: Elderly women residing in the eastern half of Baltimore, Maryland, and part of Baltimore County. PARTICIPANTS: Nine hundred nineteen moderately to severely disabled women aged 65 to 101 who participated in handgrip strength testing at baseline as part of the Women's Health and Aging Study. MEASUREMENTS: Cardiovascular disease (CVD), cancer, respiratory disease, other measures (not CVD, respiratory, or cancer), total mortality, handgrip strength, and interleukin-6. RESULTS: Over the 5-year follow-up, 336 deaths occurred: 149 due to CVD, 59 due to cancer, 38 due to respiratory disease, and 90 due to other diseases. The unadjusted relative risk (RR) of CVD mortality was 3.21 (95% confidence interval (CI) = 2.00-5.14) in the lowest and 1.88 (95% CI = 1.11-3.21) in the middle compared with the highest tertile of handgrip strength. The unadjusted RR of respiratory mortality was 2.38 (95% CI = 1.09-5.20) and other mortality 2.59 (95% CI = 1.59-4.20) in the lowest versus the highest grip strength tertile. Cancer mortality was not associated with grip strength. After adjusting for age, race, body height, and weight, the RR of CVD mortality decreased to 2.17 (95% CI = 1.26-3.73) in the lowest and 1.56 (95% CI = 0.89 2.71) in the middle, with the highest grip-strength tertile as the reference. Further adjustments for multiple diseases, physical inactivity, smoking, interleukin-6, C-reactive protein, serum albumin, unintentional weight loss, and depressive symptoms did not materially change the risk estimates. Similar results were observed for all-cause mortality. CONCLUSION: In older disabled women, handgrip strength was a powerful predictor of cause-specific and total mortality. Presence of chronic diseases commonly underlying death or the mechanisms behind decline in muscle strength in chronic disease, such as inflammation, poor nutritional status, disuse, and depression, all of which are independent predictors of mortality, did not explain the association. Handgrip strength, an indicator of overall muscle strength, may predict mortality through mechanisms other than those leading from disease to muscle impairment. Grip strength tests may help identify patients at increased risk of deterioration of health. PMID- 12752839 TI - Functional outcomes of excessive daytime sleepiness in older adults. AB - OBJECTIVES: To describe the effect of self-reported excessive daytime sleepiness (EDS) on functional outcomes. DESIGN: Case-control study designed to examine differences in functional status between cases (with daytime sleepiness) and controls (no daytime sleepiness) with regard to demographic factors, general health, sleep history, and medications. SETTING: Retirement communities in southeastern Pennsylvania, Delaware, and New Jersey. PARTICIPANTS: Seventy-six nondepressed, nondemented adults, aged 65 and older, were cases (had daytime sleepiness) and 38 were controls (had no daytime sleepiness). MEASUREMENTS: Standardized questionnaires to assess disease-specific functional status (Functional Outcomes of Sleepiness Questionnaire (FOSQ) and Epworth Sleepiness Scale (ESS)), depression (Geriatric Depression Scale-Short Form and the Center for Epidemiologic Studies-Depression Scale), dementia (Short Blessed Test), demographic factors, current medical history, and sleep complaints. RESULTS: There was a significant difference in functional status between sleepy cases and nonsleepy controls. Sleepiness had a moderate to large negative effect (effect size range from 0.59 to 0.83, P <.005) on the following functional domains of the FOSQ: social outcome, general productivity, vigilance, activity level, and global assessment of functional status. Correlation between ESS and FOSQ subscales were 0.31 to -0.67, P <.05. Examination of cases with daytime sleepiness revealed increased functional impairment in individuals with more than three medical conditions or those taking more than four medications (P <.001 and P =.03, respectively). CONCLUSION: Daytime sleepiness is associated with functional impairments in a broad range of activities. The decrease in daily functioning noted in the sleepy subjects has implications for deconditioning and related comorbidity. These findings suggest that exploration of daytime sleepiness should be part of the ongoing assessment of the elderly, particularly those with multiple medical conditions. PMID- 12752840 TI - Gastrointestinal disease control after histamine2-receptor antagonist dose modification for renal impairment in frail chronically ill elderly patients. AB - OBJECTIVES: To determine whether histamine2-receptor antagonist (H2RA) dose modified for renal impairment affects gastrointestinal (GI) disease control. DESIGN: Concurrent medical record review. SETTING: One hundred forty-six nursing facilities throughout the United States. PARTICIPANTS: Three hundred thirty-six patients aged 65 and older receiving H2RAs for GI disorders. INTERVENTION: H2RA dose modified for renal impairment or no dose change. MEASUREMENTS: Disease control (no H2RA dose increase for 6 months or longer, additional GI medication, hospitalizations, emergency room visits, and unscheduled physician visits for GI symptoms) was evaluated using chart review at 3, 6, 9, and 12 months in nursing home patients aged 65 and older with H2RA dose modified for decreased creatinine clearance (ClCr) according to manufacturer. RESULTS: Three hundred thirty-six patients, mean age +/- standard deviation 85.9 +/- 7.9, with mean ClCr of 33.6 +/ 10.4 mL/min, were recommended to receive lower H2RA doses based upon estimated renal function. Patients were analyzed in two groups: H2RA dose reduced (Group 1) and dose reduction not adopted or implemented (Group 2). There was no difference in baseline characteristics (age, weight, ClCr, or starting H2RA dose and indication) between the two groups. One hundred ninety-eight patients in Group 1 were taking 195.5 +/- 71.0 mg per day of nizatidine or equivalent, compared with 183.7 +/- 66.6 mg for 138 patients in Group 2. For patients with 90 days of follow-up, the mean H2RA dose in Group 1 was 100.2 +/- 44.3 mg, compared with 187.8 +/- 69.9 for Group 2 (P <.0001) The mean decrease in daily dose for Groups 1 and 2 after 365 days were 98.9 +/- 72.9 mg and 22.2 +/- 68.2 mg, respectively (P <.0001). Except for more physician visits in Group 2, disease control was similar for all groups. Major and minor GI bleeding events were similar across both groups and over time. The 12-month mortality rate was 12.1% and 21.7% for Groups 1 and 2, respectively. This difference was statistically significant (P =.02). CONCLUSION: The findings suggest that the dose of H2RAs may be decreased based upon renal function in frail elderly patients without compromising GI disease control. PMID- 12752841 TI - Meta-analysis of psychosocial interventions for caregivers of people with dementia. AB - OBJECTIVES: To review published reports of interventions for caregivers (CGs) of persons with dementia, excluding respite care, and provide recommendations to clinicians. DESIGN: Meta-analytical review. Electronic databases and key articles were searched for controlled trials, preferably randomized, published in English from 1985 to 2001 inclusive. Thirty studies were located and scored according to set criteria, and the interventions' research quality and clinical significance were judged. SETTING: Home or noninstitutional environment. PARTICIPANTS: Informal CGs-persons providing unpaid care at home or in a noninstitutional setting. MEASUREMENTS: The primary measures were psychological morbidity and burden. Other varied outcome measures such as CG coping skills and social support were combined with measures of psychological distress and burden to form a main outcome measure. RESULTS: The quality of research increased over the 17 years. Results from 30 studies (34 interventions) indicated, at most-current follow-up, significant benefits in caregiver psychological distress (random effect size (ES) = 0.31; 95% confidence interval (CI) = 0.13-0.50), caregiver knowledge (ES = 0.51; CI = 0.05-0.98), any main caregiver outcome measure (ES = 0.32; CI = 0.15 0.48), and patient mood (ES = 0.68; CI = 0.30-1.06), but not caregiver burden (ES = 0.09; CI = -0.09-0.26). There was considerable variability in outcome, partly because of differences in methodology and intervention technique. Elements of successful interventions could be identified. Success was more likely if, in addition to CGs, patients were involved. Four of seven studies indicated delayed nursing home admission. CONCLUSION: Some CG interventions can reduce CG psychological morbidity and help people with dementia stay at home longer. Programs that involve the patients and their families and are more intensive and modified to CGs' needs may be more successful. Future research should try to improve clinicians' abilities to prescribe interventions. PMID- 12752842 TI - Income-related differences in the use of evidence-based therapies in older persons with diabetes mellitus in for-profit managed care. AB - OBJECTIVES: To determine whether income influences evidence-based medication use by older persons with diabetes mellitus in managed care who have the same prescription drug benefit. DESIGN: Observational cohort design with telephone interviews and clinical examinations. SETTING: Managed care provider groups that contract with one large network-model health plan in Los Angeles County. PARTICIPANTS: A random sample of community-dwelling Medicare beneficiaries with diabetes mellitus aged 65 and older covered by the same pharmacy benefit. MEASUREMENTS: Patients reported their sociodemographic and clinical characteristics. Annual household income (> or =$20,000 or <$20,000) was the primary predictor. The outcome variable was use of evidence-based therapies determined by a review of all current medications brought to the clinical examination. The medications studied included use of any cholesterol-lowering medications, use of 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins) for cholesterol lowering, aspirin for primary and secondary prevention of cardiovascular disease, and angiotensin-converting enzyme (ACE) inhibitors in those with diabetic nephropathy. The influence of income on evidence-based medication use was adjusted for other patient characteristics. RESULTS: The cohort consisted of 301 persons with diabetes mellitus, of whom 53% had annual household income under $20,000. In unadjusted analyses, there were lower rates of use of all evidence-based therapies and lower rates of statin use for persons with annual income under $20,000 than for higher-income persons. In multivariate models, statin use was observed in 57% of higher-income versus 30% of lower income respondents with a history of hyperlipidemia (P =.01) and 66% of higher income versus 29% of lower-income respondents with a history of myocardial infarction (P =.03). There were no differences by income in the rates of aspirin or ACE inhibitor use. CONCLUSION: Among these Medicare managed care beneficiaries with diabetes mellitus, all of whom had the same pharmacy benefit, there were low rates of use of evidence-based therapies overall and substantially lower use of statins by poorer persons. PMID- 12752843 TI - Psychotropic prescription use by community-dwelling elderly in the United States. AB - OBJECTIVES: To examine psychotropic prescription use in community-dwelling elderly in the United States and its association with predisposing, enabling, and need factors. DESIGN: Retrospective analysis of the 1996 Medical Expenditure Survey (MEPS). SETTING: A national representative sample survey of the United States non-institutionalized population. PARTICIPANTS: Community-dwelling persons aged 65 and older participating in the MEPS. MEASUREMENTS: Psychotropic prescription use patterns and factors associated with the use of psychotropics in general as well as of individual classes, specifically antidepressants, antianxiety agents, and sedative/hypnotics. RESULTS: According to the MEPS, more than 6 million (19%) community-dwelling elderly persons used psychotropic medications in 1996. Nearly 3 million (9.1%) elderly were taking antidepressants, almost 2.5 million (7.5%) antianxiety agents, and 1.5 million (4.8%) sedative/hypnotics. Several correlates of psychotropic prescription use were identified. Enabling (e.g., prescription insurance) and need (e.g., health status) factors were found to be consistently associated with the use of antidepressant, antianxiety, and sedative/hypnotic agents. Predisposing factors such as sex, race, region, and education varied with the type of psychotropic drug class examined. CONCLUSION: Nearly one in five community-dwelling elderly persons used psychotropic medications, primarily antidepressants followed by antianxiety agents. Enabling and need factors were consistently associated with psychotropic classes examined, whereas most predisposing factors varied with the type of psychotropic drug class. PMID- 12752844 TI - Prospective evaluation of a screen for complex discharge planning in hospitalized adults. AB - OBJECTIVES: To test the predictive ability of the Probability of Repeated Admission (PRA) screen for nonroutine discharge planning (requiring new referrals for formal services). DESIGN: Prospective cohort. SETTING: Two hospitals within a large Midwestern, tertiary care, referral-based system. PARTICIPANTS: Nine hundred ninety-one hospitalized adults identified using a systematic sampling strategy. MEASUREMENTS: The PRA screen was administered, and use of nonroutine discharge planning resources and nonroutine discharge disposition were determined using observation and open record review. Prolonged length of stay was determined by comparing the actual length of stay with the combined average length of stay for diagnosis-related groupings. RESULTS: Significant differences in PRA scores existed in two of three endpoints, but the differences were small, and the ranges of scores overlapped almost completely. Using, logistic regression, items predicting use of nonroutine discharge-planning resources were self-rated health, caregiver availability, age, and sex (chi-square (chi2) = 105.7, df = 9, P <.001), accounting for 17.9% of the variability and area under receiver operating characteristic curve (AUC) of 0.74. Self-rated health and sex predicted prolonged length of stay (chi2 = 15.3, df = 5, P =.009), but only explained 2.2% of the variability, with an AUC of 0.58. The predictors of nonroutine discharge disposition were self-rated health, caregiver availability, age, sex, and diabetes mellitus (chi2 = 125.8, df = 11, P <.001), accounting for 23.0% of the variability, with an AUC of 0.79. CONCLUSION: The clinical utility of using the PRA as a screen for early identification of persons who use nonroutine discharge planning is limited, although certain individual items may be useful. PMID- 12752845 TI - Differential effects of raloxifene and estrogen on insulin sensitivity in postmenopausal women. AB - OBJECTIVES: To test the hypothesis that both raloxifene and estrogen would improve insulin sensitivity in postmenopausal women and that the magnitude of the effect would be similar for both drugs. DESIGN: Placebo-controlled, double-blind, randomized study. SETTING: The General Clinical Research Center of the University of Michigan Medical Center, a university hospital. PARTICIPANTS: Forty-four healthy postmenopausal women 73 +/- 7 years old (mean age +/- standard deviation) who were not receiving hormone replacement therapy. INTERVENTION: Eight weeks of drug therapy with randomization to raloxifene (n = 16), estrogen (n = 14), or placebo (n = 14). MEASUREMENTS: These subjects underwent a frequently sampled intravenous glucose tolerance test to determine insulin sensitivity (SI) and total and regional (central) body composition measurements by dual-energy x-ray absorptiometry at baseline and after 8 weeks of drug therapy. RESULTS: There were no statistically significant differences in age, body mass index, total or central fat mass, or SI between the three groups at baseline. The major outcome variable was SI. After 8 weeks of drug therapy, there was no significant change in SI in the placebo group or in the estrogen group and a significant decrease in SI in the raloxifene group, P =.003. CONCLUSION: In contrast to estrogen's ability to maintain insulin sensitivity, raloxifene decreases insulin sensitivity in healthy nondiabetic postmenopausal women. The clinical significance of this effect of raloxifene to impair insulin sensitivity in postmenopausal women warrants further evaluation in future studies. PMID- 12752846 TI - Screening for preexisting cognitive impairment in older intensive care unit patients: use of proxy assessment. AB - OBJECTIVES: To determine the prevalence of preexisting cognitive impairment (CI) in patients admitted to the medical intensive care unit (ICU) and compare two different proxy measures of preexisting CI in ICU patients. DESIGN: Cross sectional comparative study. SETTING: Urban university teaching hospital. PARTICIPANTS: One hundred thirty patients aged 65 and older admitted to the medical ICU. MEASUREMENTS: Two previously validated proxy measures of CI: the Modified Blessed Dementia Rating Scale (MBDRS) and the Informant Questionnaire on Cognitive Decline in the Elderly (IQCODE). RESULTS: The prevalence of preexisting CI in the ICU, determined using a combination of the measures, was 42%. Agreement between the two CI measures was 86%, with a kappa of 0.69, with discrepancies being related to the different domains measured by each instrument. CONCLUSION: There is a high prevalence of preexisting CI in patients admitted to the medical ICU. Both the MBDRS and IQCODE can be used to screen for preexisting CI in situations where direct patient assessment is not feasible. Future studies are needed to address physician recognition of CI and its effect on patient care decisions and outcomes. PMID- 12752847 TI - Validation of the five-item geriatric depression scale in elderly subjects in three different settings. AB - OBJECTIVES: To test the effectiveness of a five-item version of the Geriatric Depression Scale (GDS) for the screening of depression in community-dwelling older subjects, hospitalized older patients, and nursing home residents. DESIGN: A cross-sectional study. SETTING: A geriatric acute care ward, a geriatric outpatient clinic, and a nursing home. PARTICIPANTS: One hundred eighty-one cognitively intact older subjects. MEASUREMENT: All the participants had a comprehensive geriatric assessment including a neuropsychological evaluation by a geriatrician experienced in the management of depression. The five-item GDS was compared with the 15-item version of the GDS using the clinical diagnosis according to the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition criteria as the criterion standard. The sensitivity, the specificity, the overall accuracy, positive and negative predictive values, and positive and negative likelihood ratios were calculated. The agreement between each of two different versions of the GDS and the clinical diagnosis and the test-retest and the interrater reliability of the five-item scale were also evaluated. RESULTS: In the whole sample, 48.1% of the subjects were depressed. The five-item GDS had a sensitivity of 0.94 (0.91-0.98), a specificity of 0.81 (0.75-0.87), a positive predictive value of 0.81 (0.75-0.87), a negative predictive value of 0.94 (0.90 0.97), a positive likelihood ratio of 4.92 (4.39-5.5), and a negative likelihood ratio of 0.07 (0.06-0.08). The five-item GDS and the 15-item GDS showed a significant agreement with the clinical diagnosis of depression (kappa = 0.74 for both scales). The five-item GDS had good interrater reliability (kappa = 0.88) and test-retest reliability (kappa = 0.84). Similar values were obtained in each setting and in both sexes. CONCLUSION: The five-item GDS is as effective as the 15-item GDS for the screening of depression in cognitively intact older subjects. PMID- 12752848 TI - Exercise training as a therapy for chronic heart failure: can older people benefit? AB - Despite recent advances in pharmacological therapy, chronic heart failure remains a major cause of morbidity and mortality in older people. Studies of exercise training in younger, carefully selected patients with heart failure have shown improvements in symptoms and exercise capacity and in many pathophysiological aspects of heart failure, including skeletal myopathy, ergoreceptor function, heart rate variability, endothelial function, and cytokine expression. Data on mortality and hospitalization are lacking, and effects on everyday activity, depression, and quality of life are unclear. Exercise therapy for patients with heart failure appears to be safe and has the potential to improve function and quality of life in older people with heart failure. To realize these potential benefits, exercise programs that are suitable for older, frail people need to be established and tested in an older, frail, unselected population with comorbidities. PMID- 12752849 TI - Kaiser Permanente community partners project: improving geriatric care management practices. AB - This article describes a geriatric care management project that is testing whether geriatric care management plus a brief purchase of service (POS) intervention will lower medical costs, improve satisfaction with care, increase care plan adherence, and improve perceived quality of life. Kaiser Permanente members aged 65 and older who were eligible for geriatric care management and consented to participate in the study were randomized to one of four study groups: information and referral via mail, telephone care management, geriatric care management, or geriatric care management with POS capability. The POS intervention provides up to $2,000 of designated, paid services including in-home supportive services, transportation, respite, or medical equipment within the first 6 months of care management enrollment. Approximately 1,400 senior members were referred to the geriatric care management program, and 451 were randomly assigned to one of the four study groups. Those enrolled in the geriatric care management program were significantly more likely to be ethnic minorities and have lower income than the general Kaiser Permanente senior enrollment. Barriers encountered in implementing the POS intervention included establishing contractual agreements between Kaiser Permanente and private and community agencies, locating adequate and sufficient community agencies to provided needed services, monitoring service contracts, and delaying use of the POS benefit. PMID- 12752851 TI - Effect of statins on mortality and cardiovascular events in elderly high-risk persons. PMID- 12752852 TI - Using self-reported health status for risk adjustment and screening for disease management. PMID- 12752853 TI - Death from pneumonia in patients with progressive dementia. PMID- 12752854 TI - Failure to thrive related to metoclopramide therapy. PMID- 12752855 TI - Visual prognosis in extremely old patients with temporal (giant cell) arteritis. PMID- 12752856 TI - Recurrent colitis with different causes. PMID- 12752857 TI - The use of oral nutritional supplements and anorexigenic medications in homebound older adults. PMID- 12752858 TI - A tribute to mentors in geriatric medicine: results of a national survey of fellowship-trained geriatricians 1990-1998. PMID- 12752859 TI - Acupuncture and swallowing reflex in poststroke patients. PMID- 12752860 TI - Possible international normalized ratio elevation associated with celecoxib and warfarin in an elderly psychiatric patient. PMID- 12752863 TI - Nutrition plays a key role in HIV/AIDS care: FAO/WHO publish new manual for people living with HIV/AIDS. PMID- 12752861 TI - Transsphenoidal surgery in the elderly. PMID- 12752864 TI - Conducting a qualitative child interview: methodological considerations. AB - BACKGROUND: Studies of children have a long history, but the literature related to young children consists for the most part of studies on rather than with children and taking little account of what is regarded as significant and meaningful by children themselves. Researchers have relied almost exclusively on adults when collecting data about children's thoughts, feelings and experiences. Interviewing children, however, gives an opportunity to gain information about their subjective experiences. AIM: The purpose of this article is to illustrate the theoretical premises of child interviewing, as well as to describe some practical methodological solutions used during interviews. Factors that influence data gathered from children and strategies for taking these factors into consideration during the interview are also described. METHOD: This paper is based on literature and the experience of one of the authors in interviewing children aged from 4 to 11 years about their experiences of pain. FINDINGS AND CONCLUSION: A consideration of literature dealing with the principles of child interviewing shows that there is surprisingly little guidance available on conversational methods involving children. The empirical and conceptual foundation for child interviewing is not very clear. Novice researchers especially may need recommendations about how to conduct a qualitative child interview. The method must suit both the purpose and the context. PMID- 12752865 TI - Impact of clinical nurse specialists in multiple sclerosis--synthesis of the evidence. AB - BACKGROUND: Multiple sclerosis is a chronic neurological condition demanding a broad range of interventions and support. Multiple sclerosis nurse specialists are emerging as a leading force in providing care to this group of patients. AIM: This review aimed to identify and synthesize the evidence on the role of clinical nurse specialists in meeting the care needs of people with multiple sclerosis. METHODS: A systematic review of the literature addressing the role of the multiple sclerosis nurse specialist was undertaken. The review examined both the appropriateness and effectiveness of the multiple sclerosis nurse specialist role. The content of each item identified in the review was analysed, examining the structure, process and outcomes variables associated with the role. Materials containing an explicit methodology were critically appraised using established schedules and graded as strong, moderate or weak. The data were then synthesized in tables, thematically and using a quasi-judicial approach called the 'System of Reasoning'. FINDINGS: Fifty-five items were examined and most (53%; n = 18) were descriptive in nature. There was insufficient evidence to demonstrate that the multiple sclerosis nurse role makes a difference to care. However, evidence was found to support current descriptions of the role - meaning? and there appeared to be a good fit between the role and the care needs of people with multiple sclerosis. CONCLUSION: A systematic overview of the attributes of the multiple sclerosis nurse role is provided which should help service providers, nurses and other professionals consider how multiple sclerosis nurse specialists roles can contribute to the care of people with this condition. While there is little current evidence of effectiveness for the multiple sclerosis nurse specialist role, there is evidence for its appropriateness, although more rigorous primary research is required to test this. PMID- 12752866 TI - Tiredness and fatigue in the postnatal period. AB - AIMS OF THE PAPER: This paper aims to analyse critically a selective review of the literature on tiredness and fatigue in health and illness. This analysis is then applied to the experience of the new mother in the postnatal period. BACKGROUND AND RATIONALE: Although the twin concepts of tiredness and fatigue are well known in both sickness and health, our understanding of them is less complete. This paradox of familiarity and lack of understanding applies particularly to the postnatal period. METHOD: A literature search was undertaken using MEDLINE, CINAHL and Web of Science. The search terms 'tiredness', 'fatigue' and 'post natal' were used. FINDINGS: This search of the literature produced a small number of articles. Few of these, however, addressed the problem of tiredness or fatigue in the postnatal period, suggesting that this problem needs more research attention. CONCLUSIONS: On the basis of this analysis, it is suggested that childbirth education could be extended to foster more realistic expectations and more effective coping skills to facilitate women's adjustment to motherhood. The possibility of midwives educating mothers about the need to access, recruit and delegate some household activities to willing helpers is addressed. We propose that midwives can help women to have more realistic expectations about life after the birth, by providing advice and legitimizing the need for support and the use of coping mechanisms to assist the transition to motherhood. PMID- 12752867 TI - Effect of positioning on back pain after coronary angiography. AB - BACKGROUND: Coronary angiography is a routine cardiac diagnostic procedure in Hong Kong. Patients are restricted to bedrest after the procedure due to potential vascular complications from using a femoral approach. Many patients are required to remain on bedrest for up to 24 hours after the procedure. The effects of reducing this bedrest time is still under investigation. In the meantime, nursing interventions aimed at decreasing patient discomfort due to prolonged bedrest are feasible to implement. AIMS: The aims of this study were to evaluate the severity of back pain related to bedrest duration after coronary angiography and to compare the effects of changing patients' position in bed on their perceptions of back pain and on vascular complications. METHODS: An experimental design was used, with patients randomly assigned either to a control or experimental group. The control group received the usual care, remaining supine and flat for 8-24 hours, with the affected leg straight. The experimental group changed their body position hourly, varying between supine, right side-lying, and left side-lying during the first 7 hours after coronary angiography. RESULTS: A total of 419 patients participated in the study (control, n = 213; experimental, n = 206). Regardless of group assignment, back pain intensity increased with longer time on bedrest. In addition, the control group reported higher levels of pain at all five assessment times. Vascular complications in terms of bleeding at the femoral site were not significantly different between the control and experimental groups. CONCLUSION: The study findings suggest that patients may be able safely to change their position in bed earlier in the post-coronary angiography period than currently recommended in practice protocols. Changing position in bed may also reduce back pain, promote physical comfort, and possibly reduce patients' negative feelings toward coronary angiography. PMID- 12752868 TI - Effects of a self-management course for patients with systemic lupus erythematosus. AB - BACKGROUND: For people living with systemic lupus erythematosus, the disease's potential variety and severity of manifestations and unpredictable course present challenges and repercussions in all arenas of life. AIM: A quasi-experimental study was conducted to examine the effects of a systemic lupus erythematosus self management course for Korean patients on fatigue, coping skills, self-efficacy, depression, pain and disease activity. METHODS: In a two-group pre- and post-test design, a total of 41 participants were assigned to the experimental group (21 participants) and to the control group (20 participants). The experimental group received six weekly 2-hour sessions for groups of 10-15 literate adults of all ages, while the control group did not receive any intervention. Outcome measures included fatigue, coping skills, self-efficacy, depression, pain and disease activity. RESULTS: Patients who participated in the self-management course showed significant improvement in fatigue (P = 0.049), coping skills (P = 0.007), self efficacy (P = 0.001), and depression (P = 0.025). There were no significant changes in pain and disease activity after the intervention. CONCLUSIONS: The systemic lupus erythematosus self-management course had effects in reducing fatigue and depression and improving coping skills and self-efficacy. This course is potentially a good nursing intervention that can be offered in community settings. PMID- 12752869 TI - Converting to insulin in primary care: an exploration of the needs of practice nurses. AB - BACKGROUND: In order to optimize glycaemic control, substantial numbers of people with type 2 diabetes may require transfer from oral medication to insulin therapy. Although insulin conversion is traditionally a specialist secondary care function, as nursing roles change and expand there is growing pressure for this to be performed within primary care. However, little is known about the potential barriers to such a change, particularly from the standpoint of the frontline staff involved. AIMS: The study aimed to explore the views of practice nurses in the United Kingdom (UK) about converting diabetic patients from oral hyperglycaemic agents to injected insulin within primary care, and to investigate what structures and resources might be useful in supporting such a change. METHODS: Semi-structured interviews were conducted with 25 practice nurses, and interpreted using content analysis to extract key conceptual themes from the transcribed interview texts. FINDINGS: Most of the nurses felt that converting to insulin in primary care had considerable benefits for patients. However, issues of time, training, confidence about performing the change, and the adequacy of support systems, both for patient and nurse, emerged as the main perceived barriers to performing insulin conversions in primary care. Worries about legal and accountability issues surrounding the nurse prescribing elements were also raised. CONCLUSIONS: Where insulin conversion within primary care is being considered, it is suggested that specific training is provided for practice nurses and general practitioners, protected time is made available, and a team working approach is fostered to prevent isolation and boost patient support. Formal mentoring or supervision support for practice nurses may also help them to adapt to this new approach. LIMITATIONS: These findings are based on the views of nurses from a single UK locality, and so widespread consultation is recommended before applying them in other settings. PMID- 12752870 TI - Understanding the roles of registered general nurses and care assistants in UK nursing homes. AB - BACKGROUND: The recent government decision to fund the costs of Registered nursing time in long-term care facilities in England through the Registered Nurse Contribution to Care renders the need to distinguish the role of Registered General Nurses (RGNs) from that of Care Assistants (CAs) in nursing homes increasingly important. AIM: The objective of this qualitative study was to obtain an in-depth understanding of the main differences between the roles and functions of RGNs and CAs working in nursing homes in the United Kingdom (UK). DESIGN: Data were collected through interviews with nine RGNs and 12 CAs employed in four different nursing homes across England. FINDINGS: Our findings suggest that RGNs have difficulty defining and limiting their roles because they have all embracing roles, doing everything and anything within the home. By contrast, CAs define their role in terms of what they are not allowed to do. This difficulty in limiting their role, in addition to their sense of professional accountability for residents' care, leads RGNs to experience difficulty in delegating tasks to CAs. Both RGNs and CAs agreed that an increase in the number of assistive staff is needed to provide residents with good quality care and suggested that a measure of resident dependency would be a good method by which to determine staffing levels. CONCLUSIONS: We recommend that job descriptions that clearly define the roles and responsibilities of both RGNs and CAs are developed so that caregivers at all levels understand each others' roles and work together to co ordinate, plan and provide residents' care. PMID- 12752871 TI - Nurse managers' professional networks, psychosocial resources and self-rated health. AB - BACKGROUND: Nurse managers act under constant pressure to develop strategies in response to professional challenges within a changing and restructured health care system. When such environmental stress is present, they need access to sufficient psychosocial recourse. AIM: The study aimed to investigate whether nurse managers' professional networks, psychosocial work conditions, job support, social network and support were associated with self-rated health, sick-leave and salary. METHODS: From a total of 268 Swedish nurse managers, active in management positions, 205 (77%) agreed to participate in the study by answering a self report questionnaire. Cronbach's alpha was used to calculate internal consistency. Odds ratios were used to estimate the bivariate association between self-rated health and psychosocial resources. RESULTS: Nurse managers exposed to high job demands had significantly increased odds for low self-rated health. It was also found that low level of support from professional network, job support, social network and social support outside work displayed increased odds for low self-rated health, independently of age, gender and education. There were additive (but no synergistic) effects found for job demand in combination with professional networks, job support and emotional support and in relation to self rated health. CONCLUSION: The study showed that nurse managers exposed to high job demands had elevated odds for low self-rated health, regardless of level of psychosocial resources within or outside work. Two-thirds of the nurse managers who were affiliated to professional networks did not consider this a supporting factor in their management work. Those with low job support had increased odds for sick-leave compared with those with high support. No significant associations were found between psychosocial characteristics and salary. PMID- 12752872 TI - Evaluation of a pilot mentoring programme for nurse managers. AB - BACKGROUND: The Institute of Nursing Executives is a professional organization with a membership of approximately 150 Australian nurse managers and administrators. Members of rural zones were concerned by the lack of support available to new managers working in isolated areas and sought to develop a mentoring programme that would establish both professional development and support networks for these managers. A pilot programme was developed for two rural areas of New South Wales and one metropolitan site (the city of Sydney). AIM: The evaluation reported here aimed to determine participant expectations of mentoring and outcomes of the pilot programme. METHOD: The programme included matching and self-selection of mentor and mentee roles and attendance at a full day Mentoring Workshop. The programme was evaluated by voluntary and anonymous pre- and postworkshop questionnaires sent to all participants, and postworkshop telephone interviews. RESULTS: Thirty-seven participants (79% of those enrolled in workshop) responded to the preworkshop questionnaire (20 identifying as mentees; 17 as mentors). Findings reiterated the lack of professional support and access for rural/remote area nurse managers and illustrated that new nurse managers lacked confidence in coaching and stimulating staff. Expectations of both mentors and mentees were similar in valuing a confidential, on-going mentoring relationship. Postworkshop questionnaires (n = 16) and telephone interviews (n = 11) highlighted issues about personal choice and timing, expectations of a structured programme and making greater use of existing technology to support and maintain networks. CONCLUSION: The pilot mentoring programme was highly successful in identifying strategies for the development of a tailored and sustainable programme for newly appointed nurse managers. Provision of a highly structured and facilitated programme carries high expectations of continued external support. Nurse managers also expressed a desire to choose when, how and whom they would select as mentors. Suggestions for the future included a greater use of technology to facilitate e-mail and internet based discussion groups and mentor support. PMID- 12752873 TI - Sacrificing the personal to the professional: community mental health nurses. AB - BACKGROUND: The study of stress in mental health nurses has almost always focused on factors intrinsic to their job roles and has neglected the interaction between work and home roles, which may also be a source of stress. Moreover, role theory has not provided an adequate account of either personal or occupational change, especially when this results from the experience of stress. METHODS: Twenty community mental health nurses agreed to participate in semi-structured interviews about the stresses they experienced at work and at home. A grounded theory analysis disclosed the centrality of conflicts between work and home roles in participants' accounts and this provided the focus for subsequent detailed readings of the interview transcripts. FINDINGS: Participants described how difficult and often demanding family situations were integrated with professional careers in a climate of rapid organizational change in the health service. As well as being an ongoing source of strain, these competing role demands were often described as leading to sudden changes for individual workers, such as periods of illness or re-evaluation of their work role. To assist in making sense of this process, the term 'punctuated equilibria' was adopted to illuminate the build-up of tension between work and domestic lives, which was described by some workers as leading to a sudden reformulation of their relationship to their work roles and employing organizations. CONCLUSIONS AND LIMITATIONS: The study was small-scale and relied on retrospective self-reports. Nevertheless, it emphasized the importance of considering all the roles that individuals play and highlights the possibility that, even when staff are apparently coping with their roles at work and home, difficulties may be building up which could lead to a sudden period of absenteeism or disillusionment. PMID- 12752877 TI - Using satisfaction to measure the quality of palliative care: a review of the literature. AB - BACKGROUND: The advent of clinical governance in British health policy has placed increased demands on health care providers and practitioners to ascertain the quality of their services. Traditional indicators of quality of health care, such as death or recovery rates, are not appropriate in palliative care. Thus, it is important to establish alternative approaches to measuring the quality of palliative care services and interventions. AIMS: Satisfaction levels have been used widely in palliative care to assess quality. A literature review was conducted which aimed to explore the strengths and weaknesses of using satisfaction as an indicator of the quality of palliative care services. It also aimed to provide a solid basis upon which further work could be built. METHODS: Five electronic databases were searched using key words and phrases and key authors. Hand searches were conducted of four journals that contributed significantly to the concept of satisfaction, and reference lists of reviewed papers were scrutinized. Relevant papers were reviewed, data were extracted and these data were thematically analysed. FINDINGS: There are a number of important unresolved issues in the literature with regard to using satisfaction as an indicator of the quality of palliative care services. First, few alternatives to satisfaction are available. Secondly, satisfaction is under-theorized and no widely accepted definition exists. Thirdly, there are methodological inconsistencies across studies. It is important to take into account these findings when planning and implementing change following service evaluation using satisfaction as a measure. CONCLUSIONS: Relying on findings of satisfaction surveys to determine clinical and policy amendments in palliative care may not result in improvements in overall quality of care. Using satisfaction as a method of assessing the quality of health care services is particularly problematic and requires further investigation in both practical and conceptual terms. PMID- 12752878 TI - Clinical practice in a computer world: considering the issues. AB - BACKGROUND: In an era when information technology developments are being harnessed by governments to improve patient care, it is important to consider the issues raised by the use of computers in health care practice and policy. AIM: The aim of this paper is to provide an appraisal of considerations raised by the use of computers in health care. REVIEW FOCUS: Drawing on a range of studies in the social and medical sciences, the paper reviews the considerations. The introduction to the paper maps out the policy context, which is followed by three substantive sections: first, computer hardware and software considerations; second, issues around confidentiality; and third, personal, social and cultural considerations. CONCLUSION: This paper demonstrates that information technology developments must be allied with government or state direction and should be informed by evidence. Whilst the issues discussed are of primary relevance to the United Kingdom National Health Service, they have wider relevance to health care systems internationally. PMID- 12752879 TI - Functional status in older women following hip fracture. AB - BACKGROUND: Women aged 65 years and over are at high risk for a fractured hip because of osteoporosis. One of the devastating effects of a fracture is the loss of ability to live independently. AIM: The major aim of this study was to describe the functional status, or ability to perform activities of daily living, of women aged 65 years and older who had experienced a fractured hip in the recent past and to explore possible differences in selected variables (e.g. age and length of rehabilitation) for those who were fully performing activities of daily living at least 6 months after fracture and those who were not. DESIGN: Following Institutional Review Board approval, data were collected from 23 Caucasian women ranging from 65 to 95 years of age (M = 82, SD = 6.83) who had experienced fractured hips related to a fall in the recent past (6 months to 4 years). All were living independently prior to their fractures. Three Registered Nurses, in a structured interview, collected the data using a Demographic Data Form, an Osteoporosis Risk Factor Checklist, the Katz Index of Independence in Activities of Daily Living, and the Lawton Instrumental Activities of Daily Living Scales. RESULTS: Nine of the participants reported prior fractures, suggesting that this could have been a risk factor for this group. Eighteen achieved the maximum independence score of six on the Katz Index of Independence in Activities of Daily Living; help with bathing was required by the other five. Of the Instrumental Activities of Daily Living, those requiring physical activity for home maintenance were the most problematic. Ability to perform Instrumental Activities of Daily Living was significantly related to ability to perform the activities required to live independently. CONCLUSIONS: Results suggest that achieving a functional level that will support independence is possible for older women who were residing independently prior to a hip fracture. PMID- 12752880 TI - Negotiated care: a model for nursing work in the renal setting. AB - AIM: This article outlines a model for the nursing role in the chronic health care context of renal replacement therapy. METHODS: Materials from several streams of literature are used to conceptualize the potential for nursing work in the renal setting as negotiated care. In order to present the role of the renal nurse in this way it is contextualized by viewing the renal setting as a specialized social context constituted by a dominant professional discourse and a contrasting client discourse. DISCUSSION: While performing specific therapeutic activities in accord with the dominant discourse, renal nurses can develop a relationship with the person living on dialysis, based on responsiveness to their subjective experience reflecting the renal client discourse. In contrast to the language of noncompliance prevalent in the renal setting, nurses can, through their relationship with renal clients, facilitate their attempts to negotiate the requirements of the therapeutic regime into their own personal life situation. Nurses can mediate between the dominant and client discourses for the person living on dialysis. Care describes the quality that nurses actively seek to create in their relationships with clients, through negotiation, in order to support them to live as fully as possible while using renal replacement therapy. CONCLUSION: Within chronic health care contexts, shaped by the acute curative paradigm of biomedicine, the model of nursing work as negotiated care has the potential to humanize contemporary medical technologies by responding to clients' experiences of illness and therapy. PMID- 12752881 TI - Barriers to paediatric nurses' research utilization. AB - BACKGROUND: A number of studies have investigated barriers to research utilization among nurses in various countries, and standardized scales have been validated to study this. Reported barriers have been categorized as individual, organizational and environmental, with organizational characteristics generally accounting for more variance. However, information about research utilization among paediatric nurses is lacking. AIM: The objective of the research reported here was to investigate barriers to research utilization and relationships between those barriers and participation in research, self-reported research utilization and education among paediatric nurses. DESIGN: A survey of all nurses in a paediatric teaching hospital; 176 nurses (33.3%) responded. Two standardized measures were used, the Barriers Scale and the Edmonton Research Orientation Scale. RESULTS: Lack of time to read research was the most frequently cited barrier to using research and administrators not allowing implementation was the least frequently cited. Characteristics of the communication and of the setting were more likely to be cited as barriers to research use than were characteristics of the nurse. Nurses who reported higher levels of actual research use were slightly less likely to see characteristics within themselves as barriers. Those who had taken a course about reading or using research were more likely to see the organization as a barrier. Barriers to research use were not associated with self-reported understanding of research. CONCLUSIONS: These results are congruent with previous findings that implementing research in practice is a complex process. They indicate that individual nurses' knowledge about research may not be as important as the process by which organizations implement research. However, the Barriers Scale measures general perceptions about barriers to research utilization and not nurses' specific experiences with barriers to implementing particular research. PMID- 12752882 TI - The association between knee temperature and pain in elders with osteoarthritis of the knee: a pilot study. AB - BACKGROUND: Osteoarthritis is a highly prevalent, disabling condition that causes significant pain and suffering among older adults. Cognitively impaired elders are as susceptible to osteoarthritis as their peers. However, since they have diminished ability to communicate their pain, an alternative method to detect osteoarthritis pain in cognitively impaired elders is urgently needed. Because the late stages of osteoarthritis involve joint inflammation with a mild increase in local temperature, skin surface temperature might reasonably be expected to serve as a proxy measure of osteoarthritis pain. If knee surface temperature could be shown to predict pain in cognitively intact elders, it could be used as a proxy measure of pain for cognitively impaired elders. AIM: To test this, the study reported here assessed the relationship between knee surface temperature and pain in cognitively intact elders with osteoarthritis of the knee. METHODS: We recruited 12 cognitively intact elders with documented osteoarthritis of the knee who lived in retirement apartments. Elders' pain and knee temperature were measured three times on three separate occasions. Osteoarthritis pain of the knee was measured using the Knee Pain Scale and the Western Ontario and McMaster Osteoarthritis Index pain subscale. A YSI Model 4000 Dual Channel Display Telethermometer was used to measure knee temperature. RESULTS: We found no significant associations between knee temperature and any of the pain measures used, with one exception. However, body mass index, amount of pain medication used and activity level observed during the interview were significantly related to elders' pain. CONCLUSION: Knee temperature does not appear to predict knee pain in elders with osteoarthritis of the knee. Body mass index, use of pain medication and activity level are better predictors of this. PMID- 12752883 TI - Integration into higher education: key implementers' views on why nurse education moved into higher education. AB - AIM: The aim of this paper is to discuss the opinions of the key individuals involved in implementing the integration of nurse education into higher education in the United Kingdom (UK) about why nurse education moved into higher education and why this happened when it did. BACKGROUND: In 1995 the last of the old-style schools of nursing in the UK was fully integrated into higher education and were detached financially, legally and organizationally from District Health Authorities. However, only 6 years before, when Working Paper 10 was produced, there were only a few nursing degree courses located within higher education. What made this move into higher education particularly noteworthy was that there was never a clear statement of intent from the government that this integration of health care education was intended. Despite the fact that this is one of the most significant changes ever to take place in nurse education, there has been relatively little empirical research about why the development occurred. METHODS: A qualitative approach was selected for this study and the methods used were policy analysis and interviews. A purposeful sample of 70 implementers involved in the integration process was selected and asked for their views on this issue. FINDINGS: Participants believed that integration had occurred because of a combination of complex factors, but there was a division between those who thought that it was centrally planned and others who felt that it was an accidental outcome of the particular events of the time. CONCLUSIONS: It is not clear whether policy was influencing action or action influencing policy. Understanding of why this change occurred is needed if health care professionals wish to have greater control over future changes in education. PMID- 12752884 TI - Irish nursing students' changing self-esteem and fear of negative evaluation during their preregistration programme. AB - BACKGROUND: Studies on Irish nursing and midwifery professions have demonstrated that stress and bullying are frequent problems that may lead to depression and low self-esteem. Self-esteem is linked to social anxiety and is therefore related to fear of negative evaluation. It is important to study nursing students' feelings about self-esteem and negative evaluation, and to assess whether or not both these constructs change as students progress through their preregistration education programme. AIM: This study explored nursing students' perceived levels of self-esteem and their fear of negative evaluation prior to, and nearing the completion of, their 3-year preregistration programme. METHODS: A descriptive, quantitative, comparative survey design was used. All students in the first intake of 1995 in two general nursing schools in Southern Ireland agreed to take part (n = 72). A questionnaire developed from the Rosenberg Self-esteem Scale and the Watson and Friend Fear of Negative Evaluation Scale was used to collect data at the start of their programme and again 2 months before completion. RESULTS: In general, students' reported self-esteem rose as they neared the end of their education programme and their fear of negative evaluation decreased; however, their overall self-esteem levels at their highest were only average. CONCLUSIONS: Many of the studies examining self-esteem have produced contradictory results. An examination of the organizational factors that contribute to self-esteem may increase our understanding of the phenomenon. Self-esteem is a complex, multifaceted phenomenon. While there is no single factor that can increase or decrease a person's self-esteem, this study has explored the potential impact of the fear of negative evaluation on self-esteem. Nursing students' self-esteem might be increased by expansion of intrinsic job characteristics, improving their job satisfaction and providing frequent positive feedback. PMID- 12752885 TI - Measuring nursing workload in intensive care: an observational study using closed circuit video cameras. AB - BACKGROUND: There is an increasing demand for intensive care provision in the United Kingdom (UK), partly because of a national shortage of intensive care beds. The problem is compounded by the current method for calculating the nurse: patient ratio using a Nurse Workload Patient Category scoring system or similar adaptations used in many intensive care units. This ratio is calculated by using patient category or dependency scales, which operate on the assumption that the more critically ill the patient, the more nurse time is needed to care for the patient. However, many mechanically ventilated critically ill patients (allocated a high category of care) may need less nursing care than patients who are self ventilating and allocated a lower level of dependence. PURPOSE: In this study, a video recorder was used to document nurse activity for 48 continuous shifts in two intensive care units to determine the accuracy of the Nursing Workload Patient Category scoring system in measuring nurse workload. METHODS: The video data were correlated later with the Patient Category allocated to the patient by the nurse at the time. RESULTS: The results of this observational study demonstrated that, despite complex care needs, a high percentage of nursing activities observed in each unit consisted of low skill activity. Furthermore, nurses spent less time with patients categorized as in need of intensive care than those in need of high dependency care in both units. CONCLUSION: The findings suggest that existing nurse:patient ratio classifications may be inappropriate, since nurses spent less time with critically ill patients. Radical reconsideration of nursing levels and skill mix might make it possible to increase intensive care provision because fewer nurses would be needed to staff each bed. The findings support alternative and more flexible systems for assessing workload and the use of different nurse:patient ratios. PMID- 12752886 TI - Primary health care provision for adults with a learning disability. AB - BACKGROUND: During the last decade, primary care has been designated as the main provider of health care to people with learning disabilities. Practice nurses based in primary care teams are increasingly the first points of contact with health services. They make an important contribution to promoting good health, with health screening and illness prevention work being a significant part of their role. However, little is known about their views or involvement regarding the provision of primary care for people with learning disabilities. There is therefore a need to explore the nursing perspective, from within primary care, on the current provision of care for people with a learning disability. RESEARCH AIM: To inform the learning disability service of the role of primary care in current service provision for people with learning disabilities. RESEARCH METHOD: A survey of all practice nurses currently employed by Grampian Health Board in Aberdeen was conducted in June 2001. An initial pilot study was conducted in primary care and learning disability services. Data collection took place during June and July 2001. All practice nurses working in the geographical region of Grampian were invited to participate. RESULTS: Communication barriers exist, preventing access to health screening and treatment for some people with learning disabilities. Independent living skills, for example dietary management and budgeting, require additional community support. These conditions pose complex problems for practice nurses and other members of the primary care team. CONCLUSION: This study indicates a need for closer support and partnership with the learning disability service. The factors necessary for providing health care to people with learning disabilities extend beyond the domain of primary care teams. The findings of this survey have important implications for education and support to enable carers and professionals to provide a high standard of care. PMID- 12752888 TI - Abstracts of the annual meeting of the Australian Society for Geriatric Medicine. Darwin, Northern Territory, Australia, 24-26 June 2002. PMID- 12752887 TI - The difference between biological warfare and bioterrorism: Australia finally makes a start towards real preparedness for bioterrorism. PMID- 12752889 TI - Scleroderma renal crisis: poor outcome despite aggressive antihypertensive treatment. AB - BACKGROUND: Scleroderma renal crisis (SRC) is a rare but feared complication of scleroderma. Angiotensin--converting enzyme (ACE) inhibition has significantly improved survival, but it is unknown whether prophylactic ACE inhibitors will prevent this complication. AIMS: To determine: (i) the frequency of SRC in our cohort of well-characterized scleroderma patients resident in South Australia, (ii) any predisposing clinical and serological features, (iii) median disease duration at which SRC occurs, (iv) possible precipitants, (v) disease outcome, and (vi) whether patients were taking ACE inhibitors prior to onset of SRC. METHODS: Systematic review of the clinical course of all patients registered on the South Australian Scleroderma Register. RESULTS: SRC occurred in 16 patients. This constituted 2.8% of a total scleroderma cohort and 15% of the diffuse scleroderma cohort identified in South Australia. All 16 patients had diffuse cutaneous scleroderma. SRC occurred at a median disease duration of 15 months (range 1 week-11 years). Disease outcome was poor (five deaths, three requiring long-term dialysis and only two patients regaining a normal creatinine) despite aggressive antihypertensive treatment (including ACE inhibitors) in an intensive care or specialized renal unit. Two patients were later able to discontinue dialysis. Only two patients were taking small doses of ACE inhibitors prior to the onset of their SRC. The frequency of Scl-70 was decreased in the SRC group (P = 0.003). CONCLUSION: SRC is a rare event occurring in a small proportion of patients with diffuse scleroderma. The outcome of SRC was poor despite aggressive antihypertensive treatment. It is hypothesized that prophylactic ACE inhibition in susceptible patients might prevent or ameliorate this complication. PMID- 12752890 TI - Acute administration of conjugated equine oestrogen does not improve exercise induced myocardial ischaemia in men with coronary artery disease. AB - BACKGROUND: The parenteral administration of oestradiol acutely protects against exercise-induced myocardial ischaemia in women, but whether this effect is sex specific is not known. AIMS: The effects of acutely administered conjugated equine oestrogen on exercise-induced myocardial ischaemia in men with established coronary heart disease were investigated in a randomized, placebo-controlled, double-blind cross-over trial. METHODS: Twenty men, aged 62 +/- 11.6 years, with reproducible exercise-induced myocardial ischaemia were treated with either intravenous conjugated equine oestrogen (25 mg) or saline prior to undergoing an exercise stress test. Primary end-points were total exercise time and time to 1 mm ST-segment depression. RESULTS: All participants completed the protocol. Total exercise time exceeded the baseline value in 17 of the 20 men following saline, and in 17 of the 20 men following oestrogen pretreatment. Time to 1 mm ST-segment depression exceeded the baseline value in 14 of the 19 men following saline, and following oestrogen administration, exceeded baseline in 13 of the 19 men. There was no significant difference between the two treatments in either time to 1 mm ST-segment depression or total exercise time. A period effect was apparent for total exercise time (P = 0.05) but not for time to ST-segment depression. CONCLUSION: Acute parenteral oestrogen therapy did not increase total exercise time or time to the onset of electrocardiographic changes of ischaemia in men with chronic stable coronary artery disease. These findings contrast the favourable effects of oestrogen in women in comparable studies and indicate a sex specificity for the acute cardiovascular effects of oestrogen. PMID- 12752891 TI - Evaluation of cyclooxygenase 2 inhibitor use in patients admitted to a large teaching hospital. AB - BACKGROUND: The heavy usage of coxibs in Australia far outstrips the predicted usage that was based on the treatment of patients with risk factors for upper gastro-intestinal adverse events from conventional anti-inflammatory agents. This raises questions regarding the appropriateness of prescribing. AIMS: To determine: (i) the relationship between prescriptions for cyclooxygenase 2 (COX 2) inhibitors and objective evidence of inflammatory arthritis, (ii) prior experience with paracetamol and/or conventional non-steroidal anti-inflammatory drugs (NSAIDs), and (iii) contraindications to the use of NSAIDs. METHODS: Drug utilization evaluation and rheumatological assessment was conducted on 70 consecutive patients admitted on COX-2 inhibitors to a 480-bed metropolitan hospital. The main outcome measures were: the indication for COX-2 inhibitor; objective evidence of inflammatory arthritis; previous trial of paracetamol or conventional NSAIDs; and patient satisfaction. RESULTS: Only 11 patients (16%) had symptoms or signs of an inflammatory arthropathy, and met Pharmaceutical Benefits Schedule criteria for prescribing a COX-2 inhibitor. Fifty-nine patients (84%) had chronic osteoarthritis, degenerative spinal disease, injury or malignancy, without overt active inflammation. Fourteen patients (20%) had trialled regular paracetamol prior to using any NSAID treatment. Conventional NSAIDs had been previously used by 51 patients (73%). Eleven patients (16%) reported previous adverse gastrointestinal effects from conventional NSAIDs. On the basis of significant renal impairment (creatinine clearance <50 mL/min) or cardiac failure, cautions or contraindications applied to the use of any form of NSAID in 49% of patients. Fifteen patients (21%) had demonstrable worsening of their renal function after commencement of a COX-2 inhibitor. Thirty-one percent of patients considered the drug was effective (rated >5/10). CONCLUSIONS: Drug utilization data indicate that COX-2 inhibitors are frequently used first line for degenerative osteoarthritis in the absence of overt inflammation, without prior adequate trial of paracetamol and with disregard for the cautions and contraindications of these agents. These findings may explain the unprecedented Pharmaceutical Benefits Schedule expenditure on COX-2 inhibitors in Australia. PMID- 12752892 TI - Managing antituberculosis drug therapy by therapeutic drug monitoring of rifampicin and isoniazid. AB - BACKGROUND: Current therapeutic regimens with rifampicin and isoniazid have proven successful in treating tuberculosis, however, toxicity, therapeutic failure, relapse and multiple drug resistance are serious concerns. Optimizing drug dose using therapeutic drug monitoring (TDM) may be a better approach than administering therapy as a standard dose. AIMS: To establish and evaluate a TDM service to optimize rifampicin and isoniazid therapy. METHODS: A TDM service for rifampicin and isoniazid was established in November 1998. Drug concentration data were collected, with relevant information to interpret the results. The reason for the request, information on concomitant drug administration and a questionnaire to assess clinical response to the drug results were also obtained. RESULTS: Ninety patient episodes were accepted for study. The rifampicin plasma concentrations showed significant scatter, with 46% of the rifampicin concentrations below the normal range and 2% above the normal range. Similarly, 48% of isoniazid concentrations were below the lower target of the normal range and 29% were above the upper normal limit. There was a greater proportion of isoniazid concentrations above the normal range in female patients. CONCLUSION: Significant pharmacokinetic variability was observed for rifampicin and isoniazid in the patient population studied. Further, a substantial number of plasma concentrations fell outside the suggested normal range for both drugs. Isoniazid plasma concentrations were significantly higher in female patients compared with male patients. Despite these abnormal results, the dose of rifampicin and isoniazid was altered in only 17% of patients, however, many patients received follow-up education because of the drug result. The service was considered valuable by 83% of respondents to the questionnaire. While TDM of rifampicin and isoniazid is a valuable tool to optimize the dose of these drugs in some patients, there is an urgent need for concentration-effect studies and possibly education on the principles and practice of TDM for these drugs. PMID- 12752893 TI - Update on mobile phones and health. PMID- 12752894 TI - Chronic renal insufficiency and renoprotective strategies. AB - The increasing incidence in the number of patients with end-stage renal disease is a major public health and economic concern. There is growing awareness that early detection and intervention can have a significant impact on delaying the progression of chronic renal insufficiency and the associated comorbidities. By implementing renoprotective measures early, even modest improvements can equate to significant benefits in the long term, hence the importance of identifying at risk patients and early referrals to specialist multidisciplinary teams. In this review, some of the evidence-based renoprotective measures available are examined. PMID- 12752895 TI - Biological weapons preparedness: the role of physicians. AB - The real risk posed by biological weapons was demonstrated with the distribution of anthrax spores via the USA postal service in 2001. This review outlines the central roles of physicians in optimizing biopreparedness in Australia, including maintaining awareness of the risk, promptly recognizing an event, notifying appropriate authorities upon suspicion of an event, and instituting appropriate management. Management aspects covered include appropriate diagnostic tests, infection control procedures, and empirical therapy of agents considered possible biological weapons. The critical role of physicians as public health advocates working to prevent the use of biological weapons is also outlined. PMID- 12752896 TI - Adherence to antiretroviral therapy: are we doing enough? AB - Adherence to antiretroviral therapy is a powerful predictor of response to therapy. For optimal antiretroviral therapy response, individuals need to take more than 95% of their prescribed medication. The most widely used method for measuring adherence is self-report of the number of missed doses and this should be done at every clinic visit. There are several well-recognized predictors of poor adherence, such as illicit drug use, depression, limited knowledge or ambivalence about starting treatment. Adherence can be improved by addressing these issues or through other means such as pill boxes or electronic reminders. PMID- 12752897 TI - Questioning the decision-making capacity of surrogates. AB - When patients are unable to make medical decisions for themselves due to cognitive impairment, surrogate decision makers are often called on to guide the medical team. Important to any decision made on behalf of the patient is that the decision reflects the values and preferences of the patient in light of the patient's clinical status and prognosis. Challenges arise for the medical team when surrogates themselves have questionable decision-making capacity due to psychosocial issues, conflict of interest, or the obvious projection of their own personal values and treatment preferences instead of the patient's. Even if an alternative surrogate is available, there is no consensus on when and how to switch from the primary surrogate to the alternative surrogate. This paper uses a clinical case to explore the notion of surrogate decision-making capacity, offering guidance for determining when it is appropriate to defer to an alternative surrogate, as well as guidance for managing the process of appointing the alternative as the new primary decision maker. PMID- 12752898 TI - Transcatheter treatment of Lutembacher syndrome. PMID- 12752899 TI - Endobronchial mantle cell lymphoma. PMID- 12752900 TI - Ventricular non-compaction: a rare cause of heart failure. PMID- 12752901 TI - Building a unified nursing language system: the ICNP. PMID- 12752904 TI - Armed conflict: consequences and challenges for nurses. PMID- 12752905 TI - Why an ICNP? Links among quality, information and policy. AB - Work has been underway for over a decade to develop and use the International Classification for Nursing Practice (ICNP). The International Council of Nurses has released ICNP Alpha and Beta versions. This article is an adaptation of the presentation given at the International Council of Nurses Congress on June 12th, 2001, in Copenhagen, Denmark. When the ICNP was originally drafted, many uses were envisioned. Key among these were that the information generated would be used by clinicians, researchers, administrators and policy makers. This article introduces examples of these uses. PMID- 12752906 TI - International comparison of quality indicators in United States, Icelandic and Canadian nursing facilities. AB - AIM: To discuss the results of a comparison using minimum data set (MDS)-based quality indicators (QIs) for residents in nursing facilities in three countries (Iceland; Ontario, Canada; and Missouri, United States) together with implications regarding nursing practices and resident outcomes in these countries. METHOD: Data were extracted from databases in each country for four consecutive quarterly periods during 1997 and 1998. All facilities investigated had the required consecutive quarterly data. Analytical techniques were matched to measure resident outcomes using the same MDS-based QIs in the three countries. RESULTS: Similarities among the three countries included the use of nine or more multiple medications, weight loss, urinary tract infection, dehydration, and behavioural symptoms that affect others. Differences among the three countries included bowel and bladder incontinence, indwelling catheter use, fecal impaction, tube feeding use, development of pressure ulcers, bedridden residents, physical restraint use, depression without receiving antidepressant therapy, residents with depression, use of anti-anxiety or hypnotic drugs, use of anti psychotic drugs in the absence of psychotic and related conditions, residents spending little or no time in activities, and falls. CONCLUSIONS: Comparisons highlighted differences in clinical practices among countries, which may account for differences in resident outcomes. Learning from each other's best practices can improve the quality of care for older people in nursing homes in many countries. PMID- 12752907 TI - Patients' autonomy in surgical care: a comparison of nurses' perceptions in five European countries. AB - BACKGROUND: This report forms part of the results of an international comparative study funded by the European Commission (1998-2001). AIM: To describe and compare the maintenance of patients' autonomy on surgical wards, from the point of view of nursing staff, in five European countries (Finland, Spain, Greece, Germany and Scotland). Autonomy is defined in terms of information received and decision making by patients. METHOD: The data were collected using a questionnaire specifically designed for use in this study. Responses (response rate 66%) were obtained from 1280 nurses working on surgical wards. Data analysis was based on descriptive statistics, t-tests, analysis of variance (ANOVA) with posthoc Tukey's HSD test and logistic regression. RESULTS: There were clear between country differences in nurses' perceptions, especially on a north-south axis (Finland and Scotland vs. Greece and Spain), regarding the extent to which the autonomy of surgical patients is supported by nursing staff. Training and ethics education, in particular, were associated with nurses' perceptions of the maintenance of patient autonomy in Finland and Greece. CONCLUSION: Further research is needed to establish whether the results obtained are caused by differences in cultures, nursing practices or roles of health-care personnel or patients in different European countries. PMID- 12752908 TI - Decision-making preferences towards surgical intervention in a Hong Kong Chinese population. AB - BACKGROUND: Economic, social, ethical and legal concerns are motivating changes in western-type health practice to encourage individuals to participate in health care decisions. Patient participation in health care is perceived to be beneficial because health care is negotiated with the patient so the selected regimen of care is more likely to be acceptable. Hong Kong, in accordance with contemporary trends in other industrialized countries, has adopted a Patient's Charter that outlines patient's rights regarding health treatment and choices. Studies indicate that not all patients wish to take part in decision making about treatment. AIM: To explore the degree to which a Hong Kong Chinese population desired to participate in decision making about surgical intervention. METHOD: 96 participants, 49 surgical and medical patients and 47 students of nursing, participated. Participants were given three hypothetical scenarios: cardiac failure, where major surgery is suggested; cholecystitis, where routine surgery is suggested; and the presence of a benign skin lesion, where minor surgery is suggested. Participants were asked about the degree to which they desired to participate in the decision-making process. There were five choices, ranging from the doctor having full control, to various types of collaborative decision making, to the individual having full control. RESULTS: When the medical condition is not severe, participants desired greater participation in the decision-making process. Younger participants desired greater collaboration with the doctor in decision making while older participants preferred the doctor to have the greater input with respect to decision making. PMID- 12752909 TI - Cross-cultural adaptation of the Nordic musculoskeletal questionnaire. AB - BACKGROUND: Reports in the literature have identified a need for internationally standardized and reliable measurements to analyse musculoskeletal symptoms. Screening of musculoskeletal disorders may serve as a diagnostic tool to evaluate the work environment. The Nordic general questionnaire is a standardized instrument used to analyse musculoskeletal symptoms in an ergonomic or occupational health context. PURPOSE: To translate and adapt a version of the Nordic general questionnaire into Brazilian Portuguese and evaluate its reliability. METHOD: The cross-cultural adaptation was performed according to internationally recommended methodology, using the following guidelines: translation; back-translation; committee review; and pretesting. First, the questionnaire was independently translated into Portuguese by two teachers and one doctor, and a consensus version was generated. Second, two other translators performed a back-translation independently from one another. This version was then submitted to a committee, consisting of six specialists in the area of knowledge of the instrument, to evaluate its equivalence to the original instrument. The final version was pretested on 20 subjects randomly selected in an outpatient clinic. Reliability was assessed by a test-retest procedure at 1 day intervals using the Kappa coefficient in a group of 40 subjects. The Kappa agreement values were calculated for each one of the four questions of the questionnaire. The agreement among the same observers was substantial, varying from 0.88 to 1, according to the Kappa values. RESULTS: these demonstrated strong agreement of the instrument, suggesting that the Brazilian version of the "Standardized Nordic Questionnaire" offers substantial reliability. PMID- 12752910 TI - What do women think about menopause? A qualitative study of women's expectations, apprehensions and knowledge about the climacteric period. AB - AIM: To identify and describe expectations, apprehensions and knowledge about the menopausal period and climacteric symptoms. METHOD: Data were collected by semi structured interviews/discussions with a convenience sample of 39 women, all 47 years of age. Data interpretation and analysis were based on content analysis, but influenced by a qualitative approach. FINDINGS: These included women's expectations and feelings of freedom. Apprehensions were described as different climacteric symptoms, which were well known to the women through their own or other's experiences. The women were, to some extent, aware of the physical and psychological changes that follow the menopause. However, the women lacked knowledge about these changes or self-care activities that could prevent problems or mitigate symptoms. KEY CONCLUSIONS AND IMPLICATIONS FOR PRACTICE: Discussions on health with premenopausal women can increase their knowledge about a natural phase of life, the climacteric period. The study showed that nurses/midwives who have regular contact with some women during their life have an important role to play in providing information, as well as in the treatment of climacteric symptoms. PMID- 12752911 TI - Expectations of the child health nurse in Sweden: two perspectives. AB - BACKGROUND: the child health service exists to support and stimulate parents in order to reduce stress and to encourage an advantageous development of the preschool child. AIM: To explore and describe similarities and differences in expectations of the child health nurse, from the perspective of the recently delivered first-time mother, as compared to an expression of what the child health nurse believed mothers of infants expected of them. The data consisted of 15 interviews with child health nurses and 20 interviews with first-time mothers. Thematic content analysis resulted in seven categories of expectations. The child health nurse was expected to be someone to approach, who could assess the child's development and give immunizations and to be a supporter, counsellor, safety provider and a parent group organizer with knowledge. Similarities between the mothers' and the nurses' statements occurred more frequently than differences, which is suggested to depend on the Swedish tradition among new mothers of visiting the child health clinic. The mothers expected participation in parent groups to a higher degree than the nurses thought they did. Child health nurses who fulfil the mothers' expectations appear to require a good relationship with the mother in order to find out what she desires, which the allocation of sufficient time for regular meetings, will facilitate. Moreover, the nurse requires knowledge about children's requirements and the transition to motherhood as well as the father's important role. PMID- 12752913 TI - Oral health status in relation to ethnicity of children in the Municipality of Copenhagen, Denmark. AB - Approximately 25% of children under the age of 18 in the Municipality of Copenhagen have a non-Danish ethnic background, and it is suspected that there may be major inequalities in oral health as a result. OBJECTIVES: The objectives of this study were to describe the occurrence of dental caries in different ethnic minorities, and to analyse whether the dental caries experience of the children may be affected by cultural and behavioural differences. MATERIALS AND METHODS: The study was conducted in Copenhagen as a cross-sectional investigation of 794 children, aged 3 and 5 years old (preschool), 7 years old (Grade 1) and 15 years old (Grade 9). Children of Danish, Turkish, Pakistani, Albanian, Somali and Arabian backgrounds were selected by convenience sampling. Epidemiological data were retrieved from the Danish Recording System for the Public Dental Health Services (SCOR) and sociological data were collected by postal questionnaires. RESULTS: Marked differences in dental caries prevalence were observed when different ethnic minorities were compared to Danish children. These were most prominent for the primary dentition. At age 7, 53% of the Danish and 84% of the Albanian children were affected by dental caries, the mean caries experience was 3.5 dmfs (decayed, missed and filled surfaces) and 13.8 dmfs, respectively. Caries in incisors and/or smooth surfaces was observed in 10% of the Danish children and 48% of the Albanian children. There were cultural differences in dental attendance and self-care practices of children and parents. These socio behavioural factors may help to explain the differences in dental caries prevalence and severity. CONCLUSIONS: Development of appropriate oral health promotion strategies is urgently needed to improve oral health behaviour and attitudes of parents and children of ethnic minorities. Preventive programs should be organized at local community level in close collaboration with key persons of ethnic minority societies. PMID- 12752914 TI - The post-amalgam era: a selection of materials and their longevity in the primary and young permanent dentitions. AB - OBJECTIVES: During the last decade there has been a rapid change in the selection of dental restorative materials as the use of amalgam has decreased. The aim of this study was to obtain information on children's restorative dental care in Finland and to analyse the longevity of failed restorations. DESIGN: A random sample of public dental health care centres was drawn from the registers and the dentists working there were asked to record information for each restoration they placed during a three-day period. The survey data comprised a total of 2186 restorations in patients younger than 17 years. RESULTS: Of the children in need of restorative treatment, only a few had previous amalgam restorations. Primary caries was the main reason for restorative treatment in both primary and permanent dentitions (80% and 83%, respectively). In primary teeth, the most common restorative material was resin-modified glass ionomer cement (57.4%), whereas in permanent teeth, composite resin dominated (58.7%). Amalgam was not used at all in the primary dentition and in only 0.6% of permanent teeth. Eighteen per cent of treatments in primary and 12% in permanent teeth were replacements of previous fillings. The mean age of failed glass ionomer restorations was 2.8 years (n = 101) in the primary dentition, and 3.5 years (n = 54) in the permanent dentition. CONCLUSIONS: Until better restorative materials are developed, more attention should be paid to the prevention of dental caries as well as to the proper handling of alternative materials. PMID- 12752915 TI - Dental care of a child in pain -- a comparison of treatment planning options offered by GDPs in California and the North-west of England. AB - OBJECTIVE: To compare the treatment plans offered by general dental practitioners (GDPs) in the North-west of England (UK) and California (USA) in dealing with a child in pain. METHODS: A cross-sectional postal survey of a convenience sample of 135 UK and 170 USA GDP. These GDPs were asked to consider a case scenario of a 5-year-old child with pain from a lower first primary molar and to offer a treatment plan. The plan was to have three phases: immediate care, follow-up care and longer term preventive strategy. The influence of cost on treatment plans was also recorded. RESULTS: Response rates for the GDPs were 117 (86.6%) from the UK and 139 (81.8%) from the USA. The major differences in immediate care were as follows: 98% of Americans would take a radiograph, 41% would consider extraction, 37% would place a space maintainer and 4% would use general anaesthesia to extract a tooth. Comparable proportions for the English dentists were 20%, 21%, 2% and 40%. Later clinical options also showed differences: 39% of UK dentists would extract all first primary molars compared to only 1% of Americans. Eighty eight per cent of USA dentists would place a nickel chrome crown compared to 4% of UK respondents. There were also differences in the longer term preventive measures. Greater proportions of American dentists would offer fluoride varnish (30%) and fluoride mouthrinse (37%). Comparable UK responses were 13% and 28%. Cost of care was only mentioned by 3% of UK dentists, whereas 70% of USA dentists mentioned cost as a factor in treatment planning. CONCLUSION: There were marked differences in the treatment of a child in pain between general practitioners in the UK and the USA. Further investigations are required to elucidate the reasons for these differences. PMID- 12752916 TI - The atraumatic restorative treatment (ART) approach to restoring primary teeth in a standard dental clinic. AB - OBJECTIVES: The aims of this study are to assess the feasibility of the Atraumatic Restorative Treatment (ART) approach in primary teeth and to compare the ART approach with traditional amalgam restorations in primary molars. METHODS: This study was conducted in a paediatric dentistry clinic in Kuwait between April 1999 and December 2001. The ART and amalgam restorations were placed randomly on comparable pairs of primary molars, if available. In addition, the ART approach was used for other primary teeth that had no pulpal involvement and no perceived pain before treatment. The restorations were assessed by both the ART evaluation criteria and USPHS criteria in August-September 2000 and in August-December 2001. The mean follow-up period for the restorations was 8.3 months in the first assessment (2000) and 22 months in the second assessment (2001). The assessment was possible for 35 children (mean age = 5.7 years), 18 of whom had comparable pairs (n = 35 pairs) of restorations in their primary molars. In addition, 48 other ART restorations were assessed in 2000 and 42 in 2001. RESULTS: In a 2-year follow-up, 89.6% of all ART restorations were considered successful. The failure rate of the comparable pairs of ART and amalgam restorations was 5.7%. There was no significant difference in success rate between ART and amalgam techniques. CONCLUSIONS: Based on a 2-year follow-up evaluation, the class I ART restorations seemed to have a high success rate, indicating the appropriateness of the ART approach in primary teeth. PMID- 12752917 TI - Oral lesions and dental caries status in perinatally HIV-infected children in Northern Thailand. AB - To describe the prevalence of oral lesions and dental caries status in perinatally HIV-infected children. DESIGN: A cross-sectional study. SETTING: Paediatric HIV outpatient department at the Nakornping Provincial Hospital, Chiang Mai, Thailand. PATIENTS AND METHODS: Forty children with perinatal HIV infection, from early infancy to 12 years of age, were included in the study. These children were examined for oral lesions and dental caries. A number of children receiving antifungal and antiretroviral (ART) therapy were recorded. RESULTS: The mean DMFT and DMFS scores were both 2.1 (SD = 2.3). The dft and dfs scores were 4.1 (SD = 5.0) and 10.9 (SD = 14.8), respectively. A total of 57.5% of the children had one or more oral lesions. Oral candidiasis and hairy leukoplakia were the most common oral lesions. Only 12.5% of children had received ART. A total of 22.5% of the children had a history of receiving antifungal therapy. CONCLUSIONS: Oral lesions and dental caries were relatively high in this study. Consequently, treatment and prevention for oral lesions and dental caries are inevitably required for children with HIV infection in Northern Thailand. Furthermore, ART should be made available for all HIV-infected children to decrease the prevalence of HIV-associated oral lesions. PMID- 12752918 TI - Microbiological assessment of saliva from children subsequent to atraumatic restorative treatment (ART). AB - AIMS: The aim of this study was to evaluate mutans streptococci (MS) in the saliva following use of the atraumatic restorative treatment (ART) technique. METHODS: Sixteen 5-7-year-old children had restorations using the ART technique and employing FUJI IX glass-ionomer cement as the restorative material. Decayed tissue was manually excavated without local anaesthesia, being careful to avoid discomfort. Saliva was collected for microbiological assessment using Kit Caritest MS before treatment, one week, four weeks and one year after ART was used. The procedure for saliva collection, incubation, storage, and comparative reading of MS counts followed the manufacturer's instructions. The data were statistically analysed, using non-parametric tests (Wilcoxon Signed Ranks and Sign Test) at a significance level of 0.05. RESULTS: The results showed a significant reduction of MS levels in saliva when comparing the results before treatment with those obtained one week (95.95%; P = 0.003), four weeks (93.27%; P = 0.000) and one year (95.56%; P = 0.002) after ART. CONCLUSIONS: It is concluded from the results that the ART technique proved satisfactory and appeared to have produced a significant and sustained reduction in levels of MS. These results need to be confirmed in a larger study. PMID- 12752919 TI - Accidentally induced periodontitis in primary dentition: longitudinal examinations of periodontal bacteria and clinical conditions. AB - Periodontitis is very rare in the primary dentition although it can be accidentally induced in young children. The purpose of this study was to describe a case of periodontitis in a 4-year-old child, which was accidentally induced by insertion of small plastic tubes into the dental cervix of the primary incisors during play, and to discuss the clinical, radiographic, and microbiological findings. Removal of the plastic tubes resulted in resolution of the periodontal condition. Prevotella nigrescens and Campylobacter rectus were detected in subgingival plaque samples from the affected teeth at every examination, whereas Porphyromonas gingivalis, Treponema denticola, Prevotella intermedia, and Bacteroides forsythus were never found during the two-year observation period. The clinical and microbiological results suggest that the type of periodontitis caused by such an incidence is not progressive, unlike other periodontal diseases such as prepubertal and juvenile periodontitis. PMID- 12752920 TI - The conservative approach to managing unerupted lower premolars -- two case reports. AB - General dental practitioners frequently refer patients with unerupted premolars for specialist management. The frequency of unerupted lower second premolars in 15-year-old children has been cited as high as 9.7%. Two cases are discussed involving unilateral unerupted premolars, which initially appear to be in unfavourable positions. The first patient was referred at 16 years of age and presented with an unerupted lower left first premolar positioned along the lower border of the mandible. The second patient presented with an unerupted distally inclined, horizontally positioned second premolar impacting on the roots of the first permanent molar. Both cases were reviewed without any treatment, and both premolars erupted into excellent positions. This raises important questions concerning the possible treatment options for such teeth as well as the timing of any interceptive treatment. In cases where premolars are unerupted or impacted, a multidisciplinary approach is indicated involving orthodontics, paedodontics and oral surgery to establish the optimal treatment plan. PMID- 12752921 TI - The role of magnets in the management of unerupted teeth in children and adolescents. AB - This case report describes the use of magnets in the management of teeth that fail to erupt. Eight children aged between 10 and 15 years were treated. Magnetic traction was applied to two premolars and six molars. Seven teeth (one premolar and six molars) erupted successfully (mean treatment time with magnetic traction: 7.5 months). One premolar failed to erupt; serial radiographic assessment over a 9-month period revealed no evidence of movement and so the magnetic fixture was removed. Histological evaluation of tissue samples taken from around the fixture revealed no evidence of abnormal pathology. PMID- 12752922 TI - Compound odontoma associated with an unerupted and dilacerated maxillary primary central incisor in a young patient. AB - The occurrence of odontoma in the primary dentition is uncommon. There are very few reports of non-eruption of a dilacerated primary tooth in the literature. A rare case of compound odontoma in association with an unerupted dilacerated maxillary primary incisor in a young patient is reported. There was also a developing supernumerary tooth in the vicinity of the odontoma. The clinical features and management of these conditions are discussed. The relevant literature is reviewed. A possible causal relationship between odontoma and dilaceration is highlighted. PMID- 12752923 TI - Treatment with removable partial dentures: a longitudinal study. Part I. AB - The aim of this study was to evaluate the effectiveness of an oral rehabilitation by removable partial denture (RPD). Between 1983 and 1994, 629 patients were provided with an RPD at the Dental School of the Universite catholique de Louvain. All the RPDs were constructed with a cobalt-chromium framework. All the treatments were provided by dental students under the supervision of clinical instructors. At recall time (1998-2000), 269 patients could not be reached neither by telephone nor by mail and 27 had died. Consequently, 333 patients were called for clinical examination and 254 of these (76.3%) actually attended. For a total of 292 RPDs fitted for these 254 patients, 218 (74.7%) were still being worn at the time of the check-up. Seventy-four dentures were considered to be 'failures', either because they were replaced by another RPD or by a complete denture, or because they had actually never been worn. The statistical analysis (Mantel-Haenszel and Kaplan-Meier) shows that the number of failures is significantly higher at the lower jaw compared with the upper jaw. Most of the failures are attributable to RPDs with free-end saddles and, in particular, to class I mandibular dentures. The patients are wearing their denture(s) mostly continuously (63.6%) and award a high degree of satisfaction to their denture. In general, the results recorded may be considered as very satisfactory, all the more so as we have no regular recall procedures established at our school and as check-up asked for spontaneously by the patients in the course of the period of observation are most of the time occasional or non-existent. PMID- 12752924 TI - Treatment with removable partial dentures: a longitudinal study. Part II. AB - The aim of this study was to evaluate the effectiveness of an oral rehabilitation by removable partial denture (RPD). Between 1983 and 1994, 629 patients were provided with an RPD at the Dental School of the Universite catholique de Louvain. All the RPDs were constructed with a cobalt-chromium framework. All the treatments were provided by dental students under the supervision of clinical instructors. At recall time (1998-2000), 269 patients could not be reached neither by telephone nor by mail and 27 had died. Consequently, 333 patients were called for clinical examination and 254 of these (76.3%) actually attended. For a total of 292 RPDs checked 1893 persistent teeth were listed, with 804 serving as abutments; more than half (57.6%) of these were crowned, for which 51 debondings were recorded. For the period of observation we listed 79 abutments lost (37 at the upper jaw and 42 at the lower jaw), i.e. 9.8% of the total number of abutments. No relation could be established between the type of the abutments (natural or crowned teeth) and the losses observed (Test of Mantel-Haenszel: P=0.9496). In the upper jaw, the percentage of abutments lost was significantly higher in the presence of free-end edentulous areas as compared with bounded edentulous areas (Test of Mantel-Haenszel: P=0.0002); this difference does not appear for the lower jaws (P=0.9558). If we deduct the 25 abutments related with the 11 non-worn RPDs and the 79 abutments lost, no change becomes apparent for 92.2% of the maxillary abutments and for 85.8% of the mandibular abutments. For the 1089 other teeth, we observed the loss of 40 teeth and the appearance of caries or new fillings for 95 teeth. The fractures of cast clasps represent 3.4%. PMID- 12752925 TI - Stress analysis in a post-restored tooth utilizing the finite element method. AB - This study utilized the finite element method (FEM) to predict distribution of stresses in dentin of an endodontically treated tooth, restored with cast post and cores. For this investigation an axisymmetric model of a maxillary second pre molar that included an alveolar bone was analysed. The three tooth models evaluated were Ti-Ti alloy, NiCr-AuPd alloy and Ti-NiCr alloy as post-material and crown material with porcelain. A load of 200 N at an angle of 45 degrees to the longitudinal axis was applied on the occlusal margin of each model. The tooth was assumed isotropic, homogenous and elastic. The author prepared a calculation program using fortran 77. Investigation of the stress distributions was made in five regions; namely bottom of post, top of post, cole, metal-cement interface and metal-porcelain interface. The distributions of radial and axial stresses were plotted with length of radial. PMID- 12752926 TI - Evaluation of cetylpyridinium chloride for infection control in storage solution. AB - Storage solution is used in in vitro experimental studies to prevent dehydration of teeth collected immediately after extraction and to prevent bacterial and fungal growth in the storage media. The chemical nature of the storing agent may affect the tooth structure and material properties at the tested interface. The aim of this study was to evaluate the use of 0.1% cetylpyridinium chloride (CPC) as a new storage solution and to assess the possible effect of 0.1% thymol on microleakage and bond strength. Forty extracted human teeth were collected from 10 different dental clinics. Immediately after extraction, the teeth were randomly divided and immersed in four different storage solutions. Two test solutions of 0.1% CPC (group C) and 0.1% thymol (group T) were compared with phosphate-buffered saline (Group S - positive control) and to 3% H2O2 (group H - negative control). Bond strength test and dye penetration evaluation were then carried out. The findings suggest that the use of 0.1% CPC as storage solution does not affect bond strength to enamel. However, it may increase dye penetration at the cervical margin. The effect of 0.1% thymol on shear bond strength and dye penetration is similar to the effect of phosphate-buffered saline. PMID- 12752927 TI - Stress distribution of abutments and base displacement with precision attachment- and telescopic crown-retained removable partial dentures. AB - Five types of removable partial dentures (two attachment dentures, two telescopic dentures and one clasp denture) were designed. The two attachment dentures were retained by the rigid-precision attachments with or without a stabilizing arm, and the two telescope dentures were retained with cone telescope crowns with or without cross-arch stabilization. The stresses acting on abutment teeth and denture bases and the movements of denture bases were investigated, and the influences of denture design were clarified. The stress acting on a terminal abutment tooth retained by a rigid-precision attachment or cone telescopic crown was larger than that acting on a terminal abutment tooth retained by a clasp. The attachment dentures tended to concentrate more stress at the terminal abutment tooth than did the telescopic dentures. The stress of denture base of an attachment denture and a telescopic denture was less than that of a clasp denture. There was no difference between the stresses of attachment and telescopic dentures. The displacement of the denture base tended to be less when the denture was designed with a rigid connection for the retainer and with cross arch stabilization. PMID- 12752928 TI - Influence of tea on intrinsic colour of previously bleached enamel. AB - Aim of the study was to evaluate the influence of tea applied at various time intervals after bleaching of enamel on intrinsic tooth colour. Ninety bovine specimens were distributed among six groups (A-F, n=15). The samples of group A-D were bleached with the 10% carbamide peroxide (CP) gel VivaStyle for 8 h, followed by storing in artificial saliva for the remaining period of the day. The specimens were removed from the saliva at different intervals (A: 0 min, B: 60 min, C: 240 min) and immersed in freshly prepared black tea for 10 min. Group D (bleaching, no tea), E (no bleaching, but tea) and F (no bleaching, no tea) served as controls. These procedures were repeated for 8 days. Colour was measured at baseline, after each day, and after final cleaning using the CIELab system. Then Deltab (initial b-value - final reading), DeltaL, and composite colour (DeltaE) were statistically analysed. External bleaching (A-D) led to a distinct whitening effect with lower Deltab- (=reduction in yellow) and higher DeltaL-values (=increase in brightness) compared with controls. The Deltab- and DeltaL-values of the samples A-C were not significantly different from the samples which were bleached only. No significant difference was observed comparing specimens of group A-C. It is concluded that application of tea directly after bleaching with 10% CP does not significantly effect the outcome of a bleaching treatment irrespective of the time interval elapsed between the bleaching procedure and the contact of the tooth surface with tea. PMID- 12752929 TI - The dynamic range of TMJ sounds. AB - It is of clinical interest to record the amplitudes of temporomandibular joint (TMJ) sounds. The aim was to test the hypothesis that sealing the meatus, when placing a microphone in the ear canal affects such recording by increasing the sound pressure level (SPL). Bilateral recordings of 249 TMJ clickings were made from three subjects, using sampling rates of 48 or 96 kHz and 24 bits A/D conversion, with and without the ear canals sealed by Silicone putty. The peak-to peak equivalent sound pressure level (peSPL) was higher (P < 0.001) when the ear canal was sealed (range of mean differences was 8.3-24.9 dB peSPL). This means that the signal to noise ratio can be improved by sealing the meatus because the electronic noise level is not increased. Most important is that the dynamic range of the clicking sounds was 62 dB that is larger than the effective dynamic range of a 16 bits sound card. Future studies are needed to establish normative peSPL values. However, cards with at least 24 bits A/D conversion will be required, especially in patients with suspected disc displacement with reduction, where the difference in loudness between opening and closing clicking often is large. PMID- 12752930 TI - Fatigue resistance of removable orthodontic appliance reinforced with glass fibre weave. AB - The aim of this study was to measure the fatigue resistance of fibre-reinforced composite (FRC) reinforced polymeric parts of a removable orthodontic appliance beside the clasp. The effect of quantity and position of FRC-reinforcement were investigated. In addition, the influence of water storage on the fatigue properties was determined. The test specimens for eight groups (n=6) were manufactured from autopolymerizing acrylic resin. Polymethylmethacrylate pre impregnated woven glass fibre was used as reinforcement of acrylic resin specimens at the region of steel wire clasp. The test specimens of the control group were not reinforced. In the second group, the test specimens were reinforced with one fibre layer (thickness: 0.06 mm) on the tension side, and in the third and fourth group with two fibre layers. Fatigue resistance was measured by applying repeated bending force to the clasp. The highest fatigue resistance values were achieved when the test specimens were fibre-reinforced with two fibre layers. The lowest fatigue resistance values resulted when the test specimens were not reinforced (P=0.046, anova). Water storage had a tendency to decrease the fatigue resistance in all fibre reinforced test specimen groups. The results suggest that use of the woven polymer pre-impregnated glass FRC-reinforcement increases the fracture resistance of orthodontic appliance made of acrylic polymer. PMID- 12752931 TI - Use of a mandibular advancement device in obstructive sleep apnoea. AB - The present study investigated the effectiveness of an intra-oral mandibular advancement device in the treatment of patients with obstructive sleep apnoea (OSA) who could not tolerate or who had failed to comply with continuous positive airway pressure (CPAP). Thirty-five patients diagnosed by sleep study as suffering from OSA, who had either been unable to tolerate or had been non compliant with CPAP treatment, were included in the study. The subjects completed an Epworth sleep questionnaire. The subjects then had an oral appliance made. After using the appliance for 3 months, the patients repeated the questionnaires and had a repeat sleep study performed with the oral appliance in situ. Thirty one subjects completed the investigation. Mean AHI pre- and post-study were 26.64 and 24.06, respectively (P > 0.05). Mean Epworth scores pre- and post-study were 16.32 and 14.64, respectively (P > 0.05). Those patients with a pre-study AHI < 20 (n = 23), however, did significantly better with the appliance (P < 0.0001). Those patients with a pre-study AHI > 20 did not benefit from this device (P > 0.05). The main problems encountered were initial jaw discomfort in 18 patients and dry mouth in 11 patients (both of which improved with continued usage). It was concluded that the type of appliance used in this study can be recommended for those with mild OSA who are unable to tolerate CPAP. PMID- 12752933 TI - Atomic analysis and knoop hardness measurement of the cavity floor prepared by Er,Cr:YSGG laser irradiation in vitro. AB - In the present study, the compositional changes and knoop hardness of the cavity floor prepared by Er,Cr:YSGG laser irradiation was compared with that of the conventional bur cavity. Fifteen laser and 15 bur cavities were cross-sectioned, and subjected to atomic analysis by SEM-EDX and knoop hardness test. Statistical analyses were performed using the Mann-Whitney U-test; a value of P < 0.01 was considered significant. Surface characteristics of the prepared cavities were also investigated by light microscopy and scanning electron microscopy (SEM). The results showed that the quantities of Ca (Ca weight %) and P (P weight %) were increased significantly in the laser cavity floor but no significant differences were found between the Ca/P ratio and knoop hardness number of laser and bur cavities. The SEM observation revealed that the lased cavity surface was irregular and there was also the absence of a smear layer; the orifice of dentinal tubules was exposed. Er,Cr:YSGG laser device is considered as one of the most effective and safe devices for cavity preparation because of its many advantages. This includes easy delivery system, minimal thermal damage to the surrounding tissues, minimal thermal-induced changes of dental hard tissue compositions, and favourable surface characteristic. PMID- 12752932 TI - A clinical pilot study of the time-dependent composition of tooth bleaching systems. AB - This pilot study was undertaken to determine the compositional changes in tooth bleaching materials as a function of time in vivo. Ten patients were recruited and two bleaching systems were used - one a paste and the other a gel. Each material was placed in a custom bleaching tray and worn by each patient for each of four times - 15, 30, 60 and 120 min. The material was collected and chemically analysed for water by Karl Fischer titration and titrated for carbamide peroxide by the US Pharmacopoeia method. The paste material contained 18.66% water as supplied, and after 2 h this rose to between 28.6 and 64.4%. The gel material contained 2.85% water as supplied, and after 2 h this was diluted to between 28.5 and 73.4%. There was considerable difference in saliva uptake by the custom tray between patients. Most water uptake usually occurred within the first 30 min. Peroxide concentrations decreased in an approximately linear manner with time. There was a significant difference between the materials from baseline to 30 min and thereafter (P < 0.0009). This pilot study is an effective technique for chemical evaluation of bleaching materials. The effect of saliva is an important factor to consider, and is one that has hitherto not always been appropriately emphasized. PMID- 12752934 TI - Galvanic skin response of oral cancer patients during speech. AB - Severe speech difficulty is often caused after surgery of an oral cancer. Prosthetic treatment with a removable obturator prosthesis is generally provided for such patients. Although some speech ability is recovered with prosthetic treatment, patients sometimes complain of continued dissatisfaction with their speech. However, it is difficult to evaluate the dissatisfaction. Therefore, a new method for evaluation is desirable. In this study, such a new method using the galvanic skin response as the index for the dissatisfaction of the patient was developed, and its objectivity was investigated. Eleven patients with maxillary bone defects were selected. Prior to the evaluation, improvement of speech with the removable prosthesis was confirmed using the speech intelligibility test and the visual analogue scale. The electrical resistant value at pronunciation was measured with the measuring system composed with the apparatus (galvanic skin response (GSR) measuring apparatus), the personal computer program. The changes for the electrical resistant value after pronunciation were evaluated by calculating the decrease ratio at pronunciation [(the mean electrical resistance before pronunciation - the mean electrical resistance after pronunciation)/the mean electrical resistance before pronunciation]. This decrease ratio at pronunciation was defined as the index of the speech dissatisfaction of the subject. The mean values for the decrease ratio with prosthesis were significantly smaller than the values without prosthesis (P < 0.05). From the results of this study, it is suggested that the measurement of the electrical resistance change of the skin during speech could be a new method for evaluating the speech dissatisfaction of the post-oral-cancer patient. PMID- 12752935 TI - A preliminary report on a computer-assisted dental cast analysis system used for the prosthodontic treatment. AB - To accomplish computerized 3D morphological analyses of maxillary and mandibular casts with malocclusions on the same co-ordinate system, a new reference co ordinate located on soft tissue has been proposed consisting of the top of maxillary bilateral tubercles and incisive papilla on the maxillary cast. To test the validity of this co-ordinate system, the angles of the occlusal plane in this system were examined on 10 subjects with normal occlusion. In addition, to analyse maxillary and mandibular casts on the same co-ordinate system, a bite block was made under the intercuspal position. The maxillary cast was measured by 3D measuring system, then, the bite block was placed on maxillary casts, and measured similarly. To examine the position reproducibility of this method, 3D co ordinates of the apex of the buccal cusp of the mandibular pre-molar in five bite blocks were determined. The angles formed of the occlusal plane were 1.3 +/- 1.3 degrees and 0.2 +/- 1.1 degrees on the sagittal and frontal base plane, respectively. This co-ordinate system had enough stability to replace the occlusal plane. By the measurement of bite blocks, the co-ordinates of the mandibular cusp tips were determined within the deviation of 0.2 mm. PMID- 12752936 TI - An infection control protocol: effectiveness of immersion solutions to reduce the microbial growth on dental prostheses. AB - This investigation evaluated the effectiveness of an infection control protocol for cleansing and disinfecting removable dental prostheses. Sixty-four dentures were rubbed with sterile cotton swab immediately after they had been taken from patients' mouths. Samples were individually placed in the culture medium and immediately incubated at 37 +/- 2 degrees C. The dentures were scrubbed for 1 min with 4% chlorhexidine, rinsed for 1 min in sterile water and placed for 10 min in one of the following immersion solutions: 4% chlorhexidine gluconate, 1% sodium hypochlorite, Biocide (iodophors) and Amosan (alkaline peroxide). After the disinfection procedures, the dentures were immersed in sterile water for 3 min, reswabbed and the samples were incubated. All samples obtained in the initial culture were contaminated with micro-organisms. All the lower dentures immersed in Biocide showed positive growth, and the upper dentures were positive for growth in six of eight dentures. The 4% chlorhexidine gluconate, 1% sodium hypochlorite and Amosan solutions have been proved effective to reduce the growth of the micro-organisms in the 10 min immersion period. The protocol evaluated in this study seems to be a viable method to prevent cross-contamination between dental personnel and patients. PMID- 12752937 TI - Risk factors for temporomandibular joint pain in patients with disc displacement without reduction - a magnetic resonance imaging study. AB - The purpose of this study was to evaluate whether the magnetic resonance (MR) imaging variables of temporomandibular joint (TMJ) internal derangement, osteoarthrosis and/or effusion may predict the presence of pain in patients with a clinical disorder of an internal derangement type (ID)-III. The relationship between TMJ ID-III pain and TMJ internal derangement, osteoarthrosis and effusion was analysed in MR images of 84 TMJs in 42 patients with a clinical unilateral diagnosis of TMJ ID-III pain. Criteria for including a TMJ ID-III pain patient were report of orofacial pain referred to the TMJ, with the presence of unilateral TMJ pain during palpation, function and/or unassisted or assisted mandibular opening. Bilateral sagittal and coronal MR images were obtained to establish the presence or absence of TMJ internal derangement, osteoarthrosis and effusion. Using chi-square analysis for pair-wise comparison, the data showed a significant relationship between the MR imaging findings of TMJ ID-III pain and those of internal derangement (P=0.01) and effusion (P=0.00). Of the MR imaging variables considered simultaneously in the multiple logistic regression analysis, osteoarthrosis (P=0.82) and effusion (P=0.08) dropped out as non-significant in the diagnostic TMJ pain group when compared with the TMJ non-pain group. The odds ratio that a TMJ with an internal derangement type of disk displacement without reduction might belong to the pain group was strong (2.7:1) and highly significant (P=0.00). Significant increases in risk of TMJ pain occurred with 'disk displacement without reduction in combination with osteoarthrosis' (5.2:1) (P=0.00) and/or 'disk displacement without reduction in combination with osteoarthrosis and effusion' (6.6:1) (P=0.00). The results suggest that TMJ pain is related to internal derangement, osteoarthrosis and effusion. However, the data re-emphasize the aspect that these MR imaging variables may not be regarded as the unique and dominant factors in defining TMJ pain instances. PMID- 12752938 TI - Cytotoxicity of bisphenol A glycidyl methacrylate on cytochrome P450-producing cells. AB - Of the cytochrome P450 (CYP) family of carcinogen-activating enzymes, CYP3A is the major form found in human livers. The purpose of this study was to investigate the cytotoxic effects of dental resin monomers after being metabolized by CYP3A4 and CYP3A7, using a colony formation assay and a neutral red assay. Specimen wells were plated with transfected cells derived from the Chinese hamster lung at 100 cells well(-1). The experimental group consisted of CYP-producing 3A4-10 and 3A7-40 cells, while the control group consisted of non CYP-producing CR-119 cells. Bisphenol A (BPA) and bisphenol A glycidyl methacrylate (Bis-GMA) and a positive control (Aflatoxine Bl) were added separately to each well and cultured for 7 days. After cultivation, the number of the colonies was counted and IC50 values were determined. The data were statistically analysed by a Student's t-test. The resultant of IC50 values indicated that the monomers were not metabolically activated by CYP3A4 or CYP3A7 as compared with the control (P < 0.05). We also confirmed that these monomers act neither as activators nor as inhibitors of CYP3A4 and CYP3A7. PMID- 12752939 TI - The effect of simulated intrapulpal pressure on bond strength to enamel and dentine. AB - The purpose of this study was to evaluate the effect of simulated intrapulpal pressure on the shear bond strength (SBS) of a self-etching bonding system (Clearfil Liner Bond II) to both enamel and dentine surfaces. Forty-two caries free human molar teeth were randomly assigned to two equal groups (n=21). One group of 21 teeth was connected to a special pulpal pressure machine, which is developed to simulate in vivo conditions. The buccal and lingual surfaces of the teeth were prepared with a diamond wheel to create flat dentine and enamel surfaces. A self-etching bonding system was then applied under pulpal pressure and 2.5 mm diameter composite cylinders were bonded to the prepared surfaces. The samples were then stored in distilled water at room temperature for 24 h under pulpal pressure before SBS was tested. The second restored group was used as a control and the teeth were not connected to the pulpal pressure apparatus during material application and test procedures. They were stored for 24 h in distilled water at room temperature before the SBS test. Fracture analysis of the enamel and dentinal surfaces was performed using a stereomicroscope. Shear bond strength values to dentine was significantly reduced with pulpal pressure (P < 0.001). However, in the enamel, SBS was increased (P < 0.001). In the control group, the SBS values to dentine and enamel did not show any significant difference (P > 0.05). Based on these results, it can be concluded that further analysis of enamel histology and bonding mechanisms are needed. PMID- 12752940 TI - Analysis of depth of the microporosity in a nickel-chromium system alloy - effects of electrolytic, chemical and sandblasting etching. AB - The present study was designed to analyse the average depth of the microporosity of a nickel-chromium (Ni-Cr) system alloy (Verabond II). The metal surface was subject to one of the following surface treatment: (i) Electrolytic etching in nitric acid 0.5 N at a current density of 250 mA cm(-2); (ii) chemical etching with CG-Etch etchant; and (iii) Sandblasting with alumina particles 50 microm. Half of the samples were polished before the surface treatments. The depth of porosity was measured through photomicrographs (500x) with a profilometer, and the data were statistically analysed using an analysis of variance (anova) followed by Tukey's test. The conclusions were (i) Differents surface treatment of the Ni-Cr system alloy lead to different depths of microporosity; (ii) The greatest depth of porosity was observed in non-polished alloy; (iii) The greatest and identical depth of microporosity was observed following electrolytic etching and chemical etching; (iv) The least and identical depth of microporosity was observed with chemical etching and sandblasting with alumina particles 50 microm, and (v) Chemical etching showed an intermediary depth. PMID- 12753068 TI - Functional study of rat 5-HT2A receptors using antisense oligonucleotides. AB - We studied the effects in rats of a 6-day intracerebroventricular (i.c.v) infusion of four different end-capped phosphorothioate-modified antisense oligonucleotides (AOs), specifically targeting different regions of the 5 hydroxytryptamine2A (5-HT2A) receptor mRNA, on central 5-HT2A receptor expression and 5-HT2A receptor-mediated behaviours. Only one of the AOs (sequence 4), directed against the 5'-untranslated region (from + 557 to + 577), specifically affected central 5-HT2A receptor expression and receptor-mediated behaviour. This AO (sequence 4) reduced binding of the 5-HT2A agonist 1-(2,5-dimethoxy-4 [125I]iodophenyl)-2-aminopropane ([125I]DOI) up to 25% in cortical areas, as measured by quantitative autoradiography. Cortical binding of the antagonist [3H]ketanserin was not affected. As the specific AO treatment presumably affects the synthesis of new receptor, we hypothesize that this newly synthesized receptor represents the major part of the functionally active, G protein coupled receptor. A 5-day infusion of AO (sequence 4) resulted in profound inhibition of the head-twitch response (HTR) to 1-(2,5-dimethoxy-4-methylphenyl)-2-aminopropane (DOM). In contrast, treatment with vehicle, sense oligonucleotides (SOs) and other AOs (sequences 1, 2 and 3) caused an increased DOM-induced HTR as well as a spontaneous HTR. The latter was abolished by treatment with the 5-HT2 receptor antagonist, ritanserin. Systematic investigation of the surgical and infusion procedures revealed that the enhanced HTR already appeared following drilling of the skull. This wounding can probably damage the blood-brain barrier and cause a stress-induced increase in serotonergic transmission. AO (sequence 4) treatment also abolished the spontaneous HTR. AO (sequence 4) treatment allowed the identification of specific central 5-HT2A receptor-mediated behaviours in the complex serotonergic syndrome induced by tryptamine in rats. Only bilateral convulsions and body tremors were significantly inhibited. The backward locomotion, hunched back and Straub tail were not affected, nor was cyanosis, an index of vasoconstriction induced by peripheral 5-HT2A receptor activation. Labelling of central 5-HT2C receptors by [3H]mesulergine, and 5-HT2C receptor mediated anxiety were not attenuated by AO or SO treatment. Rats treated with AO (sequence 4) showed increased locomotor activity and a strong reactivity towards touching. We hypothesize that the down-regulation of functional 5-HT2A receptors may shift the balance between various 5-HT receptor subtypes. Our analysis of the behavioural consequences of AO treatment and the use of different AOs and SOs has shown that specific receptor-mediated behaviour can be identified. PMID- 12753069 TI - Melatonin increases survival and inhibits oxidative and amyloid pathology in a transgenic model of Alzheimer's disease. AB - Increased levels of a 40-42 amino-acid peptide called the amyloid beta protein (A beta) and evidence of oxidative damage are early neuropathological markers of Alzheimer's disease (AD). Previous investigations have demonstrated that melatonin is decreased during the aging process and that patients with AD have more profound reductions of this hormone. It has also been recently shown that melatonin protects neuronal cells from A beta-mediated oxidative damage and inhibits the formation of amyloid fibrils in vitro. However, a direct relationship between melatonin and the biochemical pathology of AD had not been demonstrated. We used a transgenic mouse model of Alzheimer's amyloidosis and monitored over time the effects of administering melatonin on brain levels of A beta, abnormal protein nitration, and survival of the mice. We report here that administration of melatonin partially inhibited the expected time-dependent elevation of beta-amyloid, reduced abnormal nitration of proteins, and increased survival in the treated transgenic mice. These findings may bear relevance to the pathogenesis and therapy of AD. PMID- 12753070 TI - Chronic hypoxia potentiates capacitative Ca2+ entry in type-I cortical astrocytes. AB - Prolonged hypoxia exerts profound effects on cell function, and has been associated with increased production of amyloid beta peptides (A beta Ps) of Alzheimer's disease. Here, we have investigated the effects of chronic hypoxia (2.5% O2, 24 h) on capacitative Ca2+ entry (CCE) in primary cultures of rat type I cortical astrocytes, and compared results with those obtained in astrocytes exposed to A beta Ps. Chronic hypoxia caused a marked enhancement of CCE that was observed after intracellular Ca2+ stores were depleted by bradykinin application or by exposure to thapsigargin (1 microM). Exposure of cells for 24 h to 1 microM A beta P(1-40) did not alter CCE. Enhancement of CCE was not attributable to cell hyperpolarization, as chronically hypoxic cells were significantly depolarized as compared with controls. Mitochondrial inhibition [by FCCP (10 microM) and oligomycin (2.5 microg/mL)] suppressed CCE in all three cell groups, but more importantly there were no significant differences in the magnitude of CCE in the three astrocyte groups under these conditions. Similarly, the antioxidants melatonin and Trolox abolished the enhancement of CCE in hypoxic cells. Our results indicate that chronic hypoxia augments CCE in cortical type-I astrocytes, a finding which is not mimicked by A beta P(1-40) and appears to be dependent on altered mitochondrial function. PMID- 12753071 TI - Gliap--a novel untypical L-asparaginase localized to rat brain astrocytes. AB - L-asparaginases catalyse the formation of the neuroactive amino acid L-aspartate by deamination of asparagine. The major pathophysiological significance of L asparaginase activity is in its clinical use for the treatment of acute lymphatic leukaemia and neoplasias that require asparagine and obtain it from circulating pools. Here we report the identification and characterization of Gliap, a cytosolic L-asparaginase, which is the founding member of a new group of L asparaginases in mammalia. Structural modelling suggests that Gliap is an atypical mammalian type-I asparaginase inasmuch as it harbours the active centre of a type-I glycosylasparaginase but, like plant-type asparaginases, lacks their auto-proteolytic site and, in addition, exhibits significant type-II L asparaginase enzymatic activity. Moreover, in contrast to glycosylasparaginases Gliap is enriched in the cytosolic fraction and not in lysosomes. The protein is particularly abundant in liver, testis and brain. In brain Gliap is exclusively expressed in astrocytes and prominently present in structures reminiscent of glial endfeet. These data suggest that Gliap is involved in astroglial production of the neuroactive amino acid L-aspartate. PMID- 12753072 TI - Temperature-dependent, neurotrophic factor-elicited, neuronal differentiation in adrenal chromaffin cell line immortalized with temperature-sensitive SV40 T antigen. AB - We established adrenal medullary cell lines from transgenic mice expressing an oncogene, the temperature-sensitive simian virus 40 large T-antigen, under the control of the tyrosine hydroxylase promoter. A clonal cell line, named tsAM5D, conditionally grew at a permissive temperature of 33 degrees C and exhibited the dopaminergic chromaffin cell phenotype as exemplified by the expression pattern of mRNA for catecholamine-synthesizing enzymes and secretory vesicle-associated proteins. tsAM5D cells proliferated at the permissive temperature in response to basic fibroblast growth factor (bFGF) and ciliary neurotrophic factor (CNTF). At a non-permissive temperature of 39 degrees C, bFGF and CNTF acted synergistically to differentiate tsAM5D cells into neuron-like cells. In addition, tsAM5D cells caused to differentiate by bFGF plus CNTF at 39 degrees C became dependent solely on nerve growth factor for their survival and showed markedly enhanced neurite outgrowth. In the presence of bFGF and CNTF, the morphological change induced by the temperature shift was associated with up-regulated expression of neuronal marker genes including neuron-specific enolase, growth-associated protein-43, microtubule-associated protein 2, neurofilament, and p75 neurotrophin receptor, indicating that the cells underwent neuronal differentiation. Thus, we demonstrated that tsAM5D cells could proliferate at permissive 33 degrees C, and also had the capacity to terminally differentiate into neuron-like cells in response to bFGF and CNTF when the oncogene was inactivated by shifting the temperature to non-permissive 39 degrees C. These results suggest that tsAM5D cells should be a good tool to allow a detailed study of mechanisms regulating neuronal differentiation. PMID- 12753073 TI - Ethanol consumption and serotonin-1A (5-HT1A) receptor function in heterozygous BDNF (+/-) mice. AB - Heterozygous brain-derived neurotrophic factor (BDNF) (+/-) mice display abnormalities in central serotonergic neurotransmission, develop decrements in serotonergic innervation of the forebrain, and exhibit enhanced intermale aggressiveness. As disturbances of serotonin neurotransmission are implicated in alcohol abuse and aggression, we have examined in BDNF (+/-) mice alcohol drinking behavior, as well as central 5-hydroxytryptamine (5-HT)1A receptor function at the level of 5-HT1A receptor-G protein interaction. BDNF (+/-) mice displayed increased ethanol intake in a two-bottle choice procedure. There was no difference in the preference ratio for non-alcoholic tastants (i.e. quinine or saccharin) between genotypes. In the brains of alcohol-naive mice, we measured [35S]GTP gamma S binding stimulated by the 5-HT1A receptor agonist (+/-)-8 hydroxy-2-dipropyl-aminotetralin hydrobromide (8-OH-DPAT; 1 microM). In BDNF (+/ ) versus wild-type (WT) mice, 5-HT1A receptor-stimulated [35S]GTP gamma S binding was significantly attenuated in the median raphe nucleus. There was a decrease in (+/-)8-OH-DPAT-stimulated [35S]GTP gamma S binding in the dorsal raphe, which did not reach statistical significance. In the hippocampus, 5-HT1A receptor stimulated [35S]GTP gamma S binding was significantly attenuated in BDNF (+/-) mice. 5-HT1A receptor-stimulated [35S]GTP gamma S binding was attenuated in the anterior cingulate cortex and lateral septum, although these reductions did not reach statistical significance. 5-HT1A receptor number was not different between genotypes in any area of brain examined, suggesting that 5-HT1A receptor function, specifically the capacity of the 5-HT1A receptor to activate G proteins, is attenuated in BDNF (+/-) mice. PMID- 12753074 TI - Subtype-specific coupling with ADP-ribosyl cyclase of metabotropic glutamate receptors in retina, cervical superior ganglion and NG108-15 cells. AB - Cyclic ADP-ribose (cADP-ribose) is a putative second messenger or modulator. However, the role of cADP-ribose in the downstream signals of the metabotropic glutamate receptors (mGluRs) is unclear. Here, we show that glutamate stimulates ADP-ribosyl cyclase activity in rat or mouse crude membranes of retina via group III mGluRs or in superior cervical ganglion via group I mGluRs. The retina of mGluR6-deficient mice showed no increase in the ADP-ribosyl cyclase level in response to glutamate. GTP enhanced the initial rate of basal and glutamate stimulated cyclase activity. GTP-gamma-S also stimulated basal activity. To determine whether the coupling mode of mGluRs to ADP-ribosyl cyclase is a feature common to individual cloned mGluRs, we expressed each mGluR subtype in NG108-15 neuroblastoma x glioma hybrid cells. The glutamate-induced stimulation of the cyclase occurs preferentially in NG108-15 cells over-expressing mGluRs1, 3, 5, and 6. Cells expressing mGluR2 or mGluRs4 and 7 exhibit inhibition or no coupling, respectively. Glutamate-induced activation or inhibition of the cyclase activity was eliminated after pre-treatment with cholera or pertussis toxin, respectively. Thus, the subtype-specific coupling of mGluRs to ADP-ribosyl cyclase via G proteins suggests that some glutamate-evoked neuronal functions are mediated by cADP-ribose. PMID- 12753075 TI - Glutamate transport alteration triggers differentiation-state selective oxidative death of cultured astrocytes: a mechanism different from excitotoxicity depending on intracellular GSH contents. AB - Recent evidence has been provided for astrocyte degeneration in experimental models of neurodegenerative insults associated with glutamate transport alteration. To determine whether astrocyte death can directly result from altered glutamate transport, we here investigated the effects of L-trans-pyrrolidine-2,4 dicarboxylate (PDC) on undifferentiated or differentiated cultured rat striatal astrocytes. PDC induced death of differentiated astrocytes without affecting undifferentiated astrocyte viability. Death of differentiated astrocytes was also triggered by another substrate inhibitor but not by blockers of glutamate transporters. The PDC-induced death was delayed and apoptotic, and death rate was dose and treatment duration-dependent. Although preceded by extracellular glutamate increase, this death was not mediated through glutamate receptor stimulation, as antagonists did not provide protection. It involves oxidative stress, as a decrease in glutathione contents and a dramatic raise in reactive oxygen species preceded cell loss, and as protection was provided by antioxidants. PDC induced a similar percentage of GSH depletion in the undifferentiated astrocytes, but only a slight increase in reactive oxygen species. Interestingly, undifferentiated astrocytes exhibited twofold higher basal GSH content compared with the differentiated ones, and depleting their GSH content was found to render them susceptible to PDC. Altogether, these data demonstrate that basal GSH content is a critical factor of astrocyte vulnerability to glutamate transport alteration with possible insights onto concurrent death of astrocytes and gliosis in neurodegenerative insults. PMID- 12753076 TI - Heterologous mu-opioid receptor adaptation by repeated stimulation of kappa opioid receptor: up-regulation of G-protein activation and antinociception. AB - The present study was designed to investigate the effect of repeated administration of a selective kappa-opioid receptor agonist (1S-trans)-3,4 dichloro-N-methyl-N-[2-(1-pyrrolidinyl)cyclohexyl]-benzeneacetamide hydrochloride [(-)U-50,488H] on antinociception and G-protein activation induced by mu-opioid receptor agonists in mice. A single s.c. injection of (-)U-50,488H produced a dose-dependent antinociception, and this effect was reversed by a selective kappa opioid receptor antagonist nor-binaltorphimine (nor-BNI). Furthermore, a single s.c. pre-treatment with (-)U-50,488H had no effect on the mu-opioid receptor agonist-induced antinociception. In contrast, repeated s.c. administration of ( )U-50,488H resulted in the development of tolerance to (-)U-50,488H-induced antinociception. Under these conditions, we demonstrated here that repeated s.c. injection of (-)U-50,488H significantly enhanced the antinociceptive effect of selective mu-opioid receptor agonists endomorphin-1, endomorphin-2 and [d-Ala2,N MePhe4,Gly-ol5] enkephalin (DAMGO). Using the guanosine-5'-o-(3-[35S]thio) triphosphate ([35S]GTP gamma S) binding assay, we found that (-)U-50,488H was able to produce a nor-BNI-reversible increase in [35S]GTP gamma S binding to membranes of the mouse thalamus, which has a high level of kappa-opioid receptors. Repeated administration of (-)U-50,488H caused a significant reduction in the (-)U-50,488H-stimulated [35S]GTP gamma S binding in this region, whereas chronic treatment with (-)U-50,488H exhibited the increase in the endomorphin-1-, endomorphin-2- and DAMGO-stimulated [35S]GTP gamma S bindings in membranes of the thalamus and periaqueductal gray. These results suggest that repeated stimulation of kappa-opioid receptors leads to the heterologous up-regulation of mu-opioid receptor functions in the thalamus and periaqueductal gray regions, which may be associated with the supersensitivity of mu-opioid receptor-mediated antinociception. PMID- 12753077 TI - An oestrogen membrane receptor participates in estradiol actions for the prevention of amyloid-beta peptide1-40-induced toxicity in septal-derived cholinergic SN56 cells. AB - Although oestrogen [17 beta-estradiol (E2)]-related neuroprotection has been demonstrated in different models, the involvement of non-classical oestrogen receptors (ERs) remains unexplored. Using the SN56 cholinergic cell line, we present evidence indicating that an ER associated with the plasma membrane participates in oestrogen-dependent inhibition of cell death induced by amyloid beta peptide (A beta) toxicity. Similarly to E2 alone, a 15-min exposure to estradiol-horseradish peroxidase (E-HRP) significantly reduced A beta-induced cell death. This effect was decreased by the ER antagonist ICI 182,780 as well as by MC-20 antibody directed to a region neighbouring the ligand-binding domain of ER alpha. Using confocal microscopy on unpermeabilized SN56 cells exposed to MC 20 antibody, we identified a protein at the plasma membrane level. Western blot analysis of purified SN56 cell membrane fractions using MC-20 antibody revealed the presence of one band with the same electrophoretic mobility as intracellular ER alpha. Using conjugated forms of the steroid, E-HRP and E2 conjugated to bovine serum albumin-FITC, we demonstrated by confocal microscopy that SN56 cells contain surface binding sites for E2. Binding of both conjugates was blocked by pre-incubation with E2 and decreased by either ICI 182,780 or MC-20 antibody in a concentration-dependent manner. Thus, a membrane-related ER that shares some structural homologies with ER alpha may participate in oestrogen-mediated neuroprotection. PMID- 12753078 TI - Two modes of exocytosis from synaptosomes are differentially regulated by protein phosphatase types 2A and 2B. AB - The inhibitors okadaic acid (OA), fostriecin (FOS) and cyclosporin A (CsA), were used to investigate the roles of protein phosphatases in regulating exocytosis in rat brain synaptosomes by measuring glutamate release and the release of the styryl dye FM 2-10. Depolarization was induced by 30 mM KCl, or 0.3 mM or 1 mM 4 aminopyridine (4AP). OA and FOS produced a similar partial inhibition of KCl- and 0.3 mM 4AP- evoked exocytosis in both assays, but had little effect upon exocytosis evoked by 1 mM 4AP. In contrast, CsA had no effect upon KCl- and 0.3 mM 4AP-evoked exocytosis, but significantly enhanced glutamate release but not FM 2-10 dye release evoked by 1 mM 4AP. None of the phosphatase inhibitors changed calcium signals from FURA-2-loaded synaptosomes either before or after depolarization. Pretreatment with 100 nM phorbol 12-myristate 13-acetate abolished the inhibitory effect of OA on exocytosis induced by 0.3 mM 4AP. Taken together, these results show that exocytosis from synaptosomes has a phosphatase sensitive and phosphatase-insensitive component, and that there are two modes of phosphatase-sensitive exocytosis that can be elicited by different depolarization conditions. Moreover, these two modes are differentially sensitive to phosphatase 2A and 2B. PMID- 12753079 TI - Pentylenetetrazole decreases metabolic glutamate turnover in rat brain. AB - Seizures were induced in rats by intraperitoneal injection of pentylenetetrazole (PTZ, 70 mg/kg), followed, 30 min later, by injection of [1-13C]glucose and [1,2 13C]acetate. Analyses of extracts from cortex, subcortex and cerebellum were performed using 13C magnetic resonance spectroscopy and HPLC. It could be shown that PTZ affected different brain regions differently. The total amounts of glutamate, glutamine, GABA, aspartate and taurine were decreased in the cerebellum and unchanged in the other brain regions. GABAergic neurones in the cortex and subcortex were not affected, whereas those in the cerebellum showed a pronounced decrease of GABA synthesis. However, glutamatergic neurones in all brain regions showed a decrease in glutamate labelling and in addition a decreased turnover in cerebellum. It could be shown that this decrease was in the metabolic pool of glutamate whereas release of glutamate was unaffected since glutamine labelling from glutamate was unchanged. Aspartate turnover was also decreased in all brain regions. Changes in astrocyte metabolism were not detected, indicating that PTZ had no effect on astrocyte metabolism in the early postictal stage. PMID- 12753080 TI - Pathogenic A beta induces the expression and activation of matrix metalloproteinase-2 in human cerebrovascular smooth muscle cells. AB - Cerebral amyloid angiopathy (CAA) is a major pathological feature of Alzheimer's disease and related disorders. Human cerebrovascular smooth muscle (HCSM) cells, which are intimately associated with CAA, have been used as an in vitro model system to investigate pathologic interactions with amyloid beta protein (A beta). Previously we have shown that pathogenic forms of A beta induce several pathologic responses in HCSM cells including fibril assembly at the cell surface, increase in the levels of A beta precursor, and apoptotic cell death. Here we show that pathogenic A beta stimulates the expression and activation of matrix metalloproteinase-2 (MMP-2). Furthermore, we demonstrate that the increase in MMP 2 activation is largely caused by increased expression of membrane type-1 (MT1) MMP expression, the primary MMP-2 activator. Finally, treatment with MMP-2 inhibitors resulted in increased HCSM cell viability in the presence of pathogenic A beta. Our findings suggest that increased expression and activation of MMP-2 may contribute to HCSM cell death in response to pathogenic A beta. In addition, these activities may also contribute to loss of vessel wall integrity in CAA resulting in hemorrhagic stroke. Therefore, further understanding into the role of MMPs in HCSM cell degeneration may facilitate designing therapeutic strategies to treat CAA found in AD and related disorders. PMID- 12753081 TI - Cloning, characterization and expression of two alternatively splicing isoforms of Ca2+/calmodulin-dependent protein kinase I gamma in the rat brain. AB - Ca2+/calmodulin-dependent protein kinase I (CaMKI), originally identified as a protein kinase phosphorylating synapsin I, has been shown to constitute a family of closely related isoforms (alpha, beta and gamma). Here, we have isolated and determined the complete primary structures of two alternatively splicing isoforms of CaMKI termed CaMKI gamma 1 and -gamma 2. CaMKI gamma 1 and -gamma 2 contain an identical N-terminal catalytic domain with different C-terminal regions due to the deletion of the 425-bp nucleotide sequence of CaMKI gamma 1 in CaMKI gamma 2. In vitro kinase assay has demonstrated the marked enhancement of the Ca2+/CaM dependent activity of CaMKI gamma 1 by the preincubation with Ca2+/calmodulin dependent protein kinase kinase (CaMKK), but no significant activation of CaMKI gamma 2. Northern blot analysis has demonstrated the predominant expression of CaMKI gamma in the brain. RT-PCR analysis has revealed similar expression patterns between CaMKI gamma 1 and CaMKI gamma 2 in various brain regions. In situ hybridization analysis has demonstrated that CaMKI gamma mRNA is expressed in a distinct pattern from other isoforms of CaMKI with predominant expression in some restricted brain regions such as the olfactory bulb, hippocampal pyramidal cell layer of CA3, central amygdaloid nuclei, ventromedial hypothalamic nucleus and pineal gland. In the primary hippocampal neurons and NG108-15 cells, transfected CaMKI gamma 1 and -gamma 2 are localized primarily in the cytoplasm and neurites but not in the nucleus. These findings suggest that both isoforms of CaMKI gamma may be involved in Ca2+ signal transduction in the cytoplasmic compartment of certain neuronal population. PMID- 12753082 TI - Hydrogen peroxide induces association between glyceraldehyde 3-phosphate dehydrogenase and phospholipase D2 to facilitate phospholipase D2 activation in PC12 cells. AB - Oxidative stress or signaling is widely implicated in apoptosis, ischemia and mitogenesis. Previously, our group reported that the hydrogen peroxide (H2O2) dependent activation of phospholipase D2 (PLD2) in PC12 cells is involved in anti apoptotic effect. However, the precise mechanism of PLD2 activation by H2O2 was not revealed. To find H2O2-dependent PLD2-regulating proteins, we immunoprecipitated PLD2 from PC12 cells and found that glyceraldehyde 3-phosphate dehydrogenase (GAPDH) coimmunoprecipitated with PLD2 upon H2O2 treatment. This interaction was found to be direct by in vitro reconstitution of purified GAPDH and PLD2. In vitro studies also indicated that PLD2-associated GAPDH was modified on its reactive cysteine residues. Koningic acid, an alkylator of GAPDH on catalytic cysteine residue, also increased interaction between the two proteins in vitro and enhanced PLD2 activity in PC12 cells. Blocking H2O2-dependent modification of GAPDH with 3-aminobenzamide resulted in the inhibition of the GAPDH/PLD2 interaction and attenuated H2O2-induced PLD2 activation in PC12 cells. From the results, we suggest that H2O2 modifies GAPDH on its catalytic cysteine residue not only to inactivate the dehydrogenase activity of GAPDH but also to endow GAPDH with the ability to bind PLD2 and the resulting association is involved in the regulation of PLD2 activity by H2O2. PMID- 12753083 TI - Chronic nicotine administration does not increase nicotinic receptors labeled by [125I]epibatidine in adrenal gland, superior cervical ganglia, pineal or retina. AB - Neuronal nicotinic acetylcholine receptors (nAChRs) were measured in CNS and peripheral tissues following continuous exposure to saline or nicotine hydrogen tartrate (3.3 or 10 mg/kg/day) for 14 days via osmotic pumps. Initially, binding of [3H](-)nicotine, [3H]cytisine and [3H]epibatidine to nAChRs was compared to determine the suitability of each for these kinds of studies. The predominant nAChR labeled by agonists in the cerebral cortex is an alpha 4 beta 2 subtype, whereas the predominant nicotinic receptors in the adrenal gland, superior cervical ganglia and pineal gland contain an alpha 3 subunit, and they do not bind either [3H](-)nicotine or [3H]cytisine with high affinity. In retina some nAChRs bind all three ligands with high affinity, and others appear to bind only [3H]epibatidine. Thus, only [3H]epibatidine had high enough affinity to be useful for measuring the nAChRs in all of the tissues. The receptors from nicotine treated rats were then measured using [125I]epibatidine, which has binding characteristics very similar to [3H]epibatidine. Treatment with the two doses of nicotine hydrogen tartrate increased binding sites in the cerebral cortex by 40% and 70%, respectively. In contrast, no significant changes in the density of receptor binding sites were found in the adrenal gland, superior cervical ganglia, pineal gland or retina. These data indicate that chronic administration of nicotine even at high doses does not increase all nicotinic receptor subtypes, and that receptors containing alpha 3 subunits may be particularly resistant to this nicotine-induced change. PMID- 12753084 TI - Heme oxygenase-1 and heme oxygenase-2 have distinct roles in the proliferation and survival of olfactory receptor neurons mediated by cGMP and bilirubin, respectively. AB - Heme oxygenase (HO) is implicated in protection against oxidative stress, proliferation and apoptosis in many cell types, including neurons. We utilized olfactory receptor neurons (ORNs) as a model to define the roles of HO-1 and HO-2 in neuronal development and survival, and to determine the mediators of these effects. The olfactory system is a useful model as ORNs display neurogenesis post natally and do not contain nitric oxide synthase (NOS) activity, which could confound results. HO isoforms were expressed in ORNs during embryogenesis and post-natally. Mice null for either HO-1 or HO-2 displayed decreased proliferation of neuronal precursors. However, apoptosis was increased only in HO-2 null mice. Cyclic GMP immunostaining was reduced in ORNs in both genotypes, providing direct evidence that HO mediates cGMP production in vivo. Bilirubin immunostaining was reduced only in HO-2 null mice. These roles for HO-1 and HO-2 were confirmed using detergent ablation of the epithelium to observe increased neurogenesis of ORNs after target disruption in HO null mice. Primary cultures of ORNs revealed that proliferative and survival effects of HO were mediated through cGMP and bilirubin, respectively. These results support a role for HO, the CO-cGMP signaling system and bilirubin in neurodevelopment and in response to injury. PMID- 12753085 TI - A role for semaphorins and neuropilins in oligodendrocyte guidance. AB - Oligodendrocytes develop in defined CNS regions as progenitor cells, which migrate to their final destinations, encountering soluble and membrane-bound signals that influence their differentiation and potential to myelinate axonal projections. To identify the regulatory genes that may be involved in this process, microarray analysis of developing oligodendroglia was performed. Several neural guidance genes, including members of the neuropilin (NP) and semaphorin families were detected. These findings were verified and expanded upon using RT PCR with RNA from fluorescent activated cell sorted A2B5+ oligodendrocyte progenitors and O4+ pro-oligodendrocytes isolated from in vitro and in vivo sources. RT-PCR, western and immunocytochemical analyses revealed that oligodendrocytes expressed NP1, several alternatively spliced isoforms of NP2, and a broad spectrum of both soluble (Class 3), membrane-spanning (Class 4-6), and membrane-tethered (Class 7) semaphorin ligands. Class 3 semaphorins, in a modified stripe assay, caused the collapse of oligodendrocyte progenitor growth cones, redirection of processes, and altered progenitor migration. Our data support a role for neuropilins and semaphorins in orchestrating the migration patterns of developing oligodendrocytes in the CNS. PMID- 12753086 TI - Gene profiling of hippocampal neuronal culture. AB - We performed mRNA expression profiling of mouse primary hippocampal neurones undergoing differentiation in vitro. We show that 2314 genes significantly changed expression during neuronal differentiation. The temporal resolution of our experiment (six time points) permits us to distinguish between gene expression patterns characteristic for the axonal and for the dendritic stages of neurite outgrowth. Cluster analysis reveals that, in the process of in vitro neuronal differentiation, a high level of expression of genes involved in the synthesis of DNA and proteins precedes the up regulation of genes involved in protein transport, energy generation and synaptic functions. We report in detail changes in gene expression for genes involved in the synaptic vesicle cycle. Data for other genes can be accessed at our website. We directly compare expression of 475 genes in the differentiating neurones and the developing mouse hippocampus. We demonstrate that the program of gene expression is accelerated in vitro as compared to the situation in vivo. When this factor is accounted for, the gene expression profiles in vitro and in vivo become very similar (median gene-wise correlation 0.787). Apparently once the cells have taken a neuronal fate, the further program of gene expression is largely independent of histological or anatomical context. Our results also demonstrate that a comparison across the two experimental platforms (cDNA microarrays and oligonucleotide chips) and across different biological paradigms is feasible. PMID- 12753087 TI - Concerted stimulation and deactivation of pertussis toxin-sensitive G proteins by chimeric G protein-coupled receptor-regulator of G protein signaling 4 fusion proteins: analysis of the contribution of palmitoylated cysteine residues to the GAP activity of RGS4. AB - Agonists stimulated high-affinity GTPase activity in membranes of HEK293 cells following coexpression of the alpha 2A-adrenoceptor and a pertussis toxin resistant mutant of Go1 alpha. Enzyme kinetic analysis of Vmax and Km failed to detect regulation of the effect of agonist by a GTPase activating protein. This did occur, however, when cells were also transfected to express RGS4. Both elements of a fusion protein in which the N-terminus of RGS4 was linked to the C terminal tail of the alpha 2A-adrenoceptor were functional, as it was able to provide concerted stimulation and deactivation of the G protein. By contrast, the alpha 2A-adrenoceptor-RGS4 fusion protein stimulated but did not enhance deactivation of a form of Go1 alpha that is resistant to the effects of regulator of G protein signaling (RGS) proteins. Employing this model system, mutation of Asn128 but not Asn88 eliminated detectable GTPase activating protein activity of RGS4 against Go1 alpha. Mutation of all three cysteine residues that are sites of post-translational acylation in RGS4 also eliminated GTPase activating protein activity but this was not achieved by less concerted mutation of these sites. These studies demonstrate that a fusion protein between a G protein-coupled receptor and an RGS protein is fully functional in providing both enhanced guanine nucleotide exchange and GTP hydrolysis of a coexpressed G protein. They also provide a direct means to assess, in mammalian cells, the effects of mutation of the RGS protein on function in circumstances in which the spatial relationship and orientation of the RGS to its target G protein is defined and maintained. PMID- 12753088 TI - MCP-1 (CCL2) protects human neurons and astrocytes from NMDA or HIV-tat-induced apoptosis. AB - Acquired immunodeficiency syndrome (AIDS)-associated dementia is often characterized by chronic inflammation, with infected macrophage infiltration of the CNS resulting in the production of human immunodeficiency virus type 1 (HIV 1) products, including tat, and neurotoxins that contribute to neuronal loss. In addition to their established role in leukocyte recruitment and activation, we identified an additional role for chemokines in the CNS. Monocyte chemoattractant protein-1 (MCP-1 or CCL2) and regulated upon activation normal T cell expressed and secreted (RANTES) were found to protect mixed cultures of human neurons and astrocytes from tat or NMDA-induced apoptosis. Neuronal and astrocytic apoptosis in these cultures was significantly inhibited by co-treatment with MCP-1 or RANTES but not IP-10. The protective effect of RANTES was blocked by antibodies to MCP-1, indicating that RANTES protection is mediated by the induction of MCP 1. The NMDA blocker, MK801, also abolished the toxic effects of both tat and NMDA. Tat or NMDA treatment of mixed cultures for 24 h resulted in increased extracellular glutamate ([Glu]e) and NMDA receptor 1 (NMDAR1) expression, potential contributors to apoptosis. Co-treatment with MCP-1 inhibited tat and NMDA-induced increases in [Glu]e and NMDAR1, and also reduced the levels and number of neurons containing intracellular tat. These data indicate that MCP-1 may play a novel role as a protective agent against the toxic effects of glutamate and tat. PMID- 12753089 TI - Molecular correlates of impaired prefrontal plasticity in response to chronic stress. AB - Disturbed adaptations at the molecular and cellular levels following stress could represent compromised neural plasticity that contributes to the pathophysiology of stress-induced disorders. Evidence illustrates atrophy and cell death of stress-vulnerable neurones in the prefrontal cortex. Reduced plasticity may be realized through the destabilized function of selective proteins involved in organizing the neuronal skeleton and translating neurotrophic signals. To elucidate the mechanisms underlying these effects, rats were exposed to chronic footshock stress. Patterns of c-fos, phospho-extracellular-regulated protein kinases 1/2 (ERK1/2), calcineurin and phospho-cyclic-AMP response-element binding protein (CREB) expression were subsequently investigated. The results indicate chronic stress-induced impairments in prefrontal and cingulate signal transduction cascades underlying neuronal plasticity. The medial prefrontal cortex, demonstrated functional hyperactivity and dendritic phospho-ERK1/2 hyperphosphorylation, while reduced c-fos and calcineurin immunoreactivity occurred in the cingulate cortex. Significantly reduced phospho-CREB expression in both cortical regions, considering its implication in brain-derived neurotrophic factor (BDNF) transcription, suggests reduced synaptic plasticity. This data confirms the damaging effect of stress on cortical activity, on a molecular level. Due to the association of these markers in the regulation of BDNF signalling, these findings suggest a central role for intracellular neurotrophin transduction members in the pathways underlying cellular actions of stress in the brain. PMID- 12753090 TI - Proteasome activation and nNOS down-regulation in neuroblastoma cells expressing a Cu,Zn superoxide dismutase mutant involved in familial ALS. AB - Reactive oxygen and nitrogen species have emerged as predominant effectors of neurodegeneration. We demonstrated that expression of the fully active G93A Cu,Zn superoxide dismutase mutant in neuroblastoma cells is associated with an increased level of oxidatively modified proteins, in terms of carbonylated residues. A parallel increase in proteasome activity was detected and this was mandatory in order to assure cell viability. In fact, proteasome inhibition by lactacystin or MG132 resulted in programmed cell death. Nitrosative stress was not involved in the oxidative unbalance, as a decrease in neuronal nitric oxide production and down-regulation of neuronal nitric oxide synthase (nNOS) level were detected. The nNOS down-regulation was correlated to increased proteolytic degradation by proteasome, because comparable levels of nNOS were detected in G93A and parental cells upon treatment with lactacystin. The altered rate of proteolysis observed in G93A cells was specific for nNOS as Cu,Zn superoxide dismutase (Cu,Zn SOD) degradation by proteasome was influenced neither by its mutation nor by increased proteasome activity. Treatment with the antioxidant 5,5'-dimethyl-1-pyrroline N-oxide resulted in inhibition of protein oxidation and decrease in proteasome activity to the basal levels. Overall these results confirm the pro-oxidant activity of G93A Cu,Zn SOD mutant and, at the same time, suggest a cross-talk between reactive oxygen and nitrogen species via the proteasome pathway. PMID- 12753091 TI - In vivo neuroprotective role of NMDA receptors against kainate-induced excitotoxicity in murine hippocampal pyramidal neurons. AB - Activation of NMDA receptors has been shown to induce either neuronal cell death or neuroprotection against excitotoxicity in cultured cerebellar granule neurons in vitro. We have investigated the effects of pretreatment with NMDA on kainate induced neuronal cell death in mouse hippocampus in vivo. The systemic administration of kainate (30 mg/kg), but not NMDA (100 mg/kg), induced severe damage in pyramidal neurons of the hippocampal CA1 and CA3 subfields 3-7 days later, without affecting granule neurons in the dentate gyrus. An immunohistochemical study using an anti-single-stranded DNA antibody and TdT mediated dUTP nick end labeling analysis both revealed that kainate, but not NMDA, induced DNA fragmentation in the CA1 and CA3 pyramidal neurons 1-3 days after administration. Kainate-induced neuronal loss was completely prevented by the systemic administration of NMDA (100 mg/kg) 1 h to 1 day previously. No pyramidal neuron was seen with fragmented DNA in the hippocampus of animals injected with kainate 1 day after NMDA treatment. The neuroprotection mediated by NMDA was prevented by the non-competitive NMDA receptor antagonist MK-801. Taken together these results indicate that in vivo activation of NMDA receptors is capable of protecting against kainate-induced neuronal damage through blockade of DNA fragmentation in murine hippocampus. PMID- 12753092 TI - The same cellular signaling pathways mediate survival in sensory neurons that switch their trophic requirements during development. AB - A distinct subpopulation of rat dorsal root sensory (DRG) neurons, termed P neurons, switch their trophic requirements for survival during development from nerve growth factor (NGF) at embryonic stages to basic fibroblast growth factor (bFGF) just after birth. We investigated in cultured P-neurons the intracellular signaling pathways mediating survival before and after this switch. The NGF induced survival was completely blocked by either wortmannin (100 nM) or PD98059 (25-50 nM), which selectively inhibit the phosphatidylinositol 3-kinase-AKT (PI3 kinase-AKT) and mitogen-activated kinase kinase extracellular regulated kinase (MEK-ERKs) pathways, respectively. NGF activated AKT and ERKs in single embryonic P-neurons, as assayed by immunofluorescence of phosphorylated proteins. In concordance with the survival assays, wortmannin and PD98059 blocked AKT and ERKs activation, respectively. Following the trophic switch, bFGF used the same signaling pathways to promote survival of post-natal P-neurons, as either wortmannin or PD98059 blocked its effect. Also, bFGF activated AKT and ERKs in single P-neurons, and this activation was blocked by the same inhibitors. These results strongly suggest that both pathways concurrently mediate the action of NGF and bFGF during embryonic and post-natal periods, respectively. Thus, we report the novel result that the switch in trophic requirements occurs with conservation of the signaling pathways mediating survival. PMID- 12753093 TI - Atomic vacancy-induced friction on the graphite surface: observation by lateral force microscopy. AB - Lateral force microscopy has been employed to investigate the frictional behaviour of atomic vacancies on the graphite surface. Such a study was only made possible by the controlled expansion of originally single-atom vacancies into multiatom vacancies, employing oxygen plasma etching for this purpose. Enhanced friction was observed on the vacancy regions compared with pristine areas of graphite, the origin of which is examined and discussed. PMID- 12753094 TI - Retention of fluorescent probes during aldehyde-free anhydrous freeze substitution. AB - Fluorescent probes are widely used for microscopy of live-cell processes, but few such probes can also be used with classically fixed or otherwise immobilized material, and none has been used without aldehyde fixation, which can introduce artefacts of structure and probe localization. Here we show that the fluorescence patterns in fungal hyphae loaded with chloromethyl aminocoumarin (CMAC), and then anhydrously freeze-substituted, without any aldehyde fixation, are similar to those seen in living hyphae. Probe loss into the mounting medium (Spurr's resin) with CMAC and five other probes tested indicated that some unwanted solubilization of probe occurred during embedding, but nevertheless vacuoles could be imaged by their retention of probe. PMID- 12753095 TI - Reducing image distortions due to temperature-related microscope stage drift. AB - Distortions in confocal 3D image data sets were related to movements of the microscope stage that matched fluctuations in laboratory temperature. Movement was apparent in all three axes. Adding a draught-proof enclosure, covering the stage, objective lenses and supporting structures, minimized these short-term fluctuations. However, the stage still tracked slow changes in laboratory temperature and was sensitive to heat sources mounted on the microscope. Suggestions are made about microscope design. PMID- 12753096 TI - A comparative study of the quantitative accuracy of three-dimensional reconstructions of spinal cord from serial histological sections. AB - We evaluated the accuracy of estimating the volume of biological soft tissues from their three-dimensional (3D) computer wireframe models, reconstructed from histological data sets obtained from guinea-pig spinal cords. We compared quantification from two methods of three-dimensional surface reconstruction to standard quantitative techniques, Cavalieri method employing planimetry and point counting and Geometric Best-Fitting. This involved measuring a group of spinal cord segments and test objects to evaluate the accuracy of our novel quantification approaches. Once a quantitative methodology was standardized there was no statistical difference in volume measurement of spinal segments between quantification methods. We found that our 3D surface reconstructions' ability to model precisely actual soft tissues provided an accurate volume quantification of complex anatomical structures as standard approaches of Cavalieri estimation and Geometric Best-Fitting. Additionally, 3D reconstruction quantitatively interrogates and three-dimensionally images spinal cord segments and obscured internal pathological features with approximately the same effort required for standard quantification alone. PMID- 12753097 TI - Measurement of the laser pulse width on the microscope objective plane by modulated autocorrelation method. AB - We report on the construction details of a compact autocorrelator set-up for the measurement of the width of infrared laser pulses at the focal plane of a microscope for two-photon excitation fluorescence imaging. One of the novelties of the set-up, which leads to an improved measurement accuracy, is the use of a modulation technique that is achieved by mounting one of the interferometer mirrors on a loudspeaker driven by a sinusoidal bias at low frequency. A non linear least-square routine selects only that part of the fluorescence signal that is modulated at the same frequency as the loudspeaker bias. To further increase the accuracy, the laser pulse width is obtained from a series of measurements at different values of the modulation bias. The autocorrelator is a compact single bread-board (10 x 20 cm); it is PC-controlled both for the acquisition and the analysis of the data and can be coupled to different ports of the microscope. The increase in the pulse width measured for three different ports of the microscope is well accounted for by the group velocity dispersion and the glass thickness of the optics found along these paths. PMID- 12753098 TI - Automated quantification and reconstruction of collagen matrix from 3D confocal datasets. AB - The geometrical structure of fibrous extracellular matrix (ECM) impacts on its biological function. In this report, we demonstrate a new algorithm designed to extract quantitative structural information about individual collagen fibres (orientation, length and diameter) from 3D backscattered-light confocal images of collagen gels. The computed quantitative data allowed us to create surface rendered 3D images of the investigated sample. PMID- 12753099 TI - Re-evaluation of differential phase contrast (DPC) in a scanning laser microscope using a split detector as an alternative to differential interference contrast (DIC) optics. AB - In this paper, differential phase imaging (DPC) with transmitted light is implemented by adding a suitable detection system to a standard commercially available scanning confocal microscope. DPC, a long-established method in scanning optical microscopy, depends on detecting the intensity difference between opposite halves or quadrants of a split photodiode detector placed in an aperture plane. Here, DPC is compared with scanned differential interference contrast (DIC) using a variety of biological specimens and objective lenses of high numerical aperture. While DPC and DIC images are generally similar, DPC seems to have a greater depth of field. DPC has several advantages over DIC. These include low cost (no polarizing or strain-free optics are required), absence of a double scanning spot, electronically variable direction of shading and the ability to image specimens in plastic dishes where birefringence prevents the use of DIC. DPC is also here found to need 20 times less laser power at the specimen than DIC. PMID- 12753106 TI - Call to action - chronic disease prevention and management. PMID- 12753100 TI - SiCf-SiBC composites: microstructural investigations of the as-received material and creep tested composites under an oxidative environment. AB - SiCf-SiBC composites fabricated by Snecma Propulsion Solide (St Medard en Jalles, France) were investigated by SEM and HRTEM in the as-received state and after creep tests performed in air, in a temperature range 1423-1573 K, under 170 and 200 MPa. These composites are reinforced by Hi-Nicalon fibres (Nippon Carbon). A pyrocarbon interphase was first deposited on the fibres. The matrix was then deposited on the fibrous preform by several chemical vapour infiltrations (CVI). As a result the matrix is multilayered and based on the Si-B-C ternary system. This matrix is self-sealing: this is due to the presence of boron inducing the formation of a sealant glass if the material is heated in an oxidative environment. This glass will protect fibres and fibre/matrix interphases against oxidation. Hi-Nicalon fibres as well as the different matrix layers were studied by HRTEM and EDX. Some investigations were carried out on the creep-tested specimens in order to characterize modifications observed in the different constituents of the composites, particularly at the interfaces between the matrix layers and at the fibre/matrix interface. It was shown that several matrix layers crystallized during the creep tests. Moreover, a thin silica layer was observed at the pyrocarbon/matrix interfaces. Differences between the behaviour of the same type of material creep tested under neutral atmosphere are discussed. PMID- 12753107 TI - Communicating and judging the quality of qualitative research: the need for a new language. AB - BACKGROUND: Traditionally UK dietitians have tended to take a more quantitative approach to research. Qualitative research which gives an in-depth view of people's experiences and beliefs is also now being used to help answer some important dietetic research questions. REVIEW: A review of the limited number of qualitative research papers in the Journal of Human Nutrition and Dietetics 1990 2002 (nine papers in all), revealed a lack of specific discussion of the quality strategies commonly used in qualitative research. This could indicate a less than robust approach, but might also reflect a different perspective on quality, or simply the difficulties associated with disseminating qualitative research to a profession whose members lack familiarity with the language. The fact that qualitative research seems to be used rarely may also indicate a poor understanding of its role. PURPOSE OF THIS PAPER: This paper seeks to clarify the potential role of qualitative research and draws on previously published guidelines for demonstrating quality. It is hoped that this will offer dietitians a framework for carrying out qualitative research and a language for reporting it, as well acting as a stimulus for discussion. PMID- 12753108 TI - Why do diabetic patients not attend appointments with their dietitian? AB - PURPOSE: Determining the prevalence of and possible reasons for nonattendance of diabetic nutritional care clinics. METHODS: Data were collected by means of a telephone survey and a review of patient records among 293 (166 attendees and 127 nonattendees) patients undergoing outpatient treatment at a university hospital. The t-tests, chi-square tests and logistic regression analysis were used to identify potential determinants of nonattendance. The theoretical framework was primarily based on the Health Belief Model. RESULTS: In univariate analysis, nonattendance at the clinic was associated with a number of factors such as not visiting other care givers, risk perceptions, body-mass index, self-rated health, health locus of control, satisfaction with the dietitian, feelings of obligation to attend, and beliefs about the effectiveness of the treatment. In multivariate analysis only health locus of control and obligation to attend the visit were significant predictors of attendance. A significant number of respondents further reported that they perceived their visits to the dietitian to be of little use. CONCLUSION: One in three diabetic patients undergoing outpatient treatment skipped one or more visits to their dietitian. Patient education to improve attendance should focus primarily on convincing patients that they can contribute to their own health, and may stress the obligation the patients have when making an appointment with the dietitian. PMID- 12753109 TI - Patients' readiness for dietary change at the beginning of counselling: a transtheoretical model-based assessment. AB - BACKGROUND: The purpose of this study was to explore patients' readiness for dietary change within a theoretical framework of the transtheoretical model. The patients were recently diagnosed to have type 2 diabetes or impaired glucose tolerance. We discuss the theoretical aspects of appropriate dietary counselling strategies from a standpoint of patient-specific stages of change. METHODS: The data included 32 audiotaped diabetes counselling sessions with 16 patients conducted by two nurses. The transcribed data was analysed by using deductive content analysis. RESULTS: The patients were at different stages of change of diabetes-affected dietary behaviour. Their stages of change varied in different dietary areas and within certain dietary habits. These stages of change could involve their overall dietary behaviour or some minor aspects of their diets. CONCLUSIONS: Understanding patient-specific stages of change orientates health counsellors to use the most appropriate counselling strategies. The transtheoretical framework helps counsellors to perceive the total range of patients' different stages of change and their effect on the implementation of counselling. However, determining patients' stages of change through examining counselling conversations is occasionally difficult. Further qualitative research is called for. PMID- 12753110 TI - Food provision and the nutritional implications of food choices made by young adult males, in a young offenders' institution. AB - The nutritional adequacy of diets provided by a prison was assessed by analysis of the kitchen menu for 1 week of a 4-week cycle. Dietary intakes were determined using a predefined 7-day diet diary in which prisoners indicated what they had eaten, and how much. A total of 159 prisoners took part in the study. The food provided by the prison kitchen was broadly in line with current dietary recommendations. Vitamin content exceeded recommendations, with the exception of niacin in the vegetarian menu (12.6 mg compared with the reference nutrient intake of 16.8 mg). Selenium content was low in all menus, but particularly in the vegetarian menu in 1997 where it equalled the lower reference nutrient intake (LRNI) (39.5 microg). Food choices made by prisoners resulted in a wide variation in dietary intakes. Fat intake (as a proportion of energy) exceeded the recommended 35% in 82% of diets in 1996, and 64% of diets in 1997. In 1996, 34% of prisoners had intakes above 40% energy as fat. High fat intakes were largely the result of consuming items from the prison shop. Vitamin D intakes were low (3.4 and 3.3 microg in 1996 and 1997, respectively) compared with the recommendation (10 microg) for those with limited exposure to sunlight. Intakes of a number of minerals fell below recommendations, with some prisoners barely meeting the LRNI. This was particularly notable for selenium where 35% of prisoners in 1996, and 60% of prisoners in 1997 had intakes below the LRNI. PMID- 12753111 TI - The estimation of food portion sizes: a comparison between using descriptions of portion sizes and a photographic food atlas by children and adults. AB - BACKGROUND: Food Photographs and standard portion sizes have been used with adults to assess portion size when recording dietary intake. The effectiveness of these methods may be reduced when memory/recall is required and children may have problems using these techniques. METHODS: Adults (47) and children (37) were recruited from amongst university personnel, their children and children's friends to assess portion sizes of nine self-served amounts of selected food items using food photographs and standard descriptions of portion sizes. Portion sizes were estimated directly after self-serving and three - 4 days later. RESULTS: Substantial differences in the estimate of portion sizes were observed for most foods regardless of the method used or the age of the subjects, median difference range: -52-100%. For children there were greater errors using both methods than for adults. Significant differences were found between the two methods of estimating weight. The food atlas provided higher median estimated weights for the majority of the food items. There were very few differences in the estimation of portion sizes between the two testing periods. CONCLUSION: The findings would suggest that either an alternative method or a modification of the methods used here for estimating portion sizes in young subjects, for example standard food portion sizes for children of different ages such as those that are being developed by the Food Standards Agency, would be more appropriate. PMID- 12753112 TI - The link between serum cholesterol and diet. PMID- 12753119 TI - Abstracts of the Society of General Internal Medicine 26th Annual Meeting. Vancouver, British Columbia, Canada. April 30-May 3, 2003. PMID- 12753141 TI - Resistance to glucocorticosteroid therapy in inflammatory bowel disease. PMID- 12753142 TI - Thermal ablative therapy for malignant liver tumors: a critical appraisal. AB - The management of primary and secondary malignant liver tumors poses a great challenge to clinicians. Although surgical resection is the gold-standard treatment, most patients have unresectable malignant liver tumors. Over the past decade, various modalities of loco-regional therapy have gained much interest. Among them, thermal ablative therapy, including cryotherapy, microwave coagulation, interstitial laser therapy, and radiofrequency ablation (RFA), have been proven to be safe and effective. Despite the effective tumor eradication achieved within cryotherapy, the underlying freeze/thaw mechanism has resulted in serious complications that include bleeding from liver cracking and the 'cryoshock' phenomenon. Thermal ablation using microwave and laser therapy for malignant liver tumors is curative and is associated with minimal complications. However, this treatment modality is effective only for tumors <3 cm diameter. Radiofrequency ablation seems to be the most promising form of thermal ablative therapy in terms of a lower complication rate and a larger volume of ablation. However, its use is restricted by the difficulty encountered when using imaging studies to monitor the areas of ablation during and after the procedure. Moreover, the techniques of RFA need to be refined in order to achieve the same oncological radicality of malignant liver tumors as achieved by surgical resection. As each of the loco-regional therapies has its own advantages and limitations, a multidisciplinary approach using a combination of therapies will be the future trend for the management of malignant liver tumors. PMID- 12753143 TI - Effects of interferon alpha therapy on the catalytic domains of the polymerase gene and basal core promoter, precore and core regions of hepatitis B virus. AB - AIMS: The aim of the present study was to examine the catalytic domains of the polymerase gene, the basal core promoter and the precore and core regions of the hepatitis B virus (HBV) genome for specific mutations. These may account for the response to interferon alpha (IFN-alpha) treatment, which may have prognostic value. METHODS: Multiple serum samples were collected prospectively from 30 patients with chronic active hepatitis B who were treated with IFN-alpha. Patients were assigned to one of three groups: group A (n = 11) and group B (n = 10) individuals were hepatitis B e antigen (HBeAg)-positive prior to treatment. Group A patients underwent HBeAg seroconversion after treatment while group B patients did not. Group C (n = 9) patients were HBeAg-negative prior to treatment. The HBV DNA was extracted from the sera collected before, during and after treatment and the various genomic regions were amplified, sequenced and examined for mutations. RESULTS: During IFN-alpha therapy, multiple changes were found in the catalytic domains of the HBV polymerase gene in all groups. The frequency of mutations and associated amino acid changes were highest in virus from group C patients and lowest in group A patients. The interdomain regions of the viral polymerase were the most affected. Multiple mutations were also found in the precore, core and core promoter regions. However, no specific mutations were associated with clinical response or outcome. CONCLUSIONS: During IFN-alpha treatment, multiple mutations occurred in the HBV genome, including the catalytic domains of the polymerase gene. Changes that did occur could not be correlated to the clinical response or treatment outcome. However, no mutations were found that have been linked to lamivudine escape, indicating that lamivudine therapy would be effective in IFN-alpha non-responder patients. PMID- 12753144 TI - Role of elevated platelet-associated immunoglobulin G and hypersplenism in thrombocytopenia of chronic liver diseases. AB - BACKGROUND AND AIM: Thrombocytopenia typically worsens with the progression of liver disease and can become a major clinical complication. Several mechanisms that contribute to thrombocytopenia have been proposed, including hypersplenism accompanied by increased platelet sequestration, platelet destruction mediated by platelet-associated immunoglobulins (PAIgG), and diminished platelet production stimulated by thrombopoietin (TPO). The purpose of the present study was to evaluate the role of each of these mechanisms in patients with liver disease associated thrombocytopenia. METHODS: Twenty-nine patients with liver cirrhosis (LC), 20 of whom were hepatitis C virus (HCV)-seropositive, 29 chronic hepatitis (CH) patients, 24 of whom were HCV-seropositive, and 16 control patients without liver or hematopoetic disease were enrolled in this study. Serum TPO levels, PAIgG, and liver-spleen volumes were determined and correlation analyses were performed. RESULTS: No differences in serum TPO levels were observed among the three groups. The PAIgG levels were significantly elevated in CH and LC patients (mean +/- SD: 56.5 +/- 42.3 and 144.6 +/- 113.6 ng/107 cells, respectively) compared with the controls (18.9 +/- 2.5 ng/107 cells, P < 0.001 for both). Spleen volume was significantly higher only in LC (428 +/- 239) compared with CH (141 +/- 55) and control (104 +/- 50 cm3) (P < 0.001), while liver volume was not significantly different between the three groups. Correlation analyses demonstrated a significant negative correlation between platelet count with PAIgG (r = - 0.517, P < 0.001) and spleen volume (r = - 0.531, P < 0.001), and no relationship between platelet count and serum TPO level (r = 0.076). CONCLUSIONS: Serum TPO level may not be directly associated with thrombocytopenia in patients with chronic hepatitis and liver cirrhosis. In contrast, spleen volume and PAIgG are associated with thrombocytopenia in such patients, suggesting that hypersplenism and immune-mediated processes are predominant thrombocytopenic mechanisms. PMID- 12753145 TI - Decreased ratio of CD4/CD8 lymphocytes might be predictive for successful interferon alpha and lamivudine combined therapy in childhood chronic hepatitis B infection: A preliminary study. AB - BACKGROUND: In the development of chronic hepatitis with hepatitis B virus infection and in response to therapy, the immune status of the infected host plays a critical role. In this study, immunological variables were assessed in patients before interferon alpha and lamivudine therapy to determine if any pretreatment immunological parameter could be an indicator of response to therapy in childhood chronic hepatitis B infection. METHODS: Forty-four patients with chronic hepatitis B virus infection, aged 9.0 +/- 3.9 years, were enrolled in the study. The pretreatment clinical features, biochemical test results, histological activity indexes and immunological parameters were evaluated. All patients received interferon alpha for 6 months and lamivudine for 52 weeks. Four patients who could not be followed up were excluded from the study. The other 40 patients have been followed for a mean period of 27.5 +/- 9.7 months after therapy discontinuation. RESULTS: Seventeen patients showed loss of hepatitis B early antigen (HBeAg) with appearance of anti-HBe (42.5%) and six of those who responded also showed loss of hepatitis B surface antigen (HBsAg) with the presence of anti-HBs (15%). Except elevated aspartate aminotransferase levels, there was no significantly correlation between response and sex, age, pretreatment duration of disease and histological activity indexes. Pretreatment immunoglobulins (Ig), IgG subclasses, complement C3, C4 and secretory IgA levels were also not found to be significantly related to response. The evaluation of lymphocyte subsets showed that therapy responders had significantly reduced pretreatment ratios of CD4/CD8+ lymphocytes due to prominent increased percentages of CD8+ cells. The other cellular immunity parameters and some cell surface adhesion molecules were similar in both groups. CONCLUSION: This study emphasizes the importance of increased pretreatment CD8+ lymphocyte percentages leading to a significant decrease in CD4/CD8 ratio in chronic hepatitis B virus infection of childhood as an immunological factor predicting response to treatment. PMID- 12753146 TI - Autonomic nervous dysfunction in patients with liver cirrhosis using 123I metaiodobenzylguanidine myocardial scintigraphy and spectrum analysis of heart rate variability. AB - BACKGROUND: It has been reported that nitric oxide (NO) synthase is induced in patients with liver cirrhosis (LC), and that an excessive production of NO enhances sympathetic nervous function. The present report describes a study of the feasibility of evaluation of abnormalities of autonomic nervous function by 123I-metaiodobenzylguanidine (MIBG) myocardial scintigraphy and heart-rate variability in patients with LC. METHODS: Low-frequency (LF) power, high frequency (HF) power, LF/HF, and 1/f fluctuations of heart rate variability were examined in 50 patients with LC (LC group), and 50 normal subjects (N group). Echocardiogram, urinary nitrite and nitrate, and cathecholamines were examined. RESULTS: Fractional shortening was observed for the hyperdynamic state of patients in the LC group according to Child's A-C classification. Washout rate of MIBG, LF/HF, and blood levels of norepinephrine increased and HF power decreased with the progression of LC. However, the urinary secretion of nitrite and nitrate were significantly increased only in cirrhotic patients with Child C. CONCLUSIONS: The present results indicate that autonomic abnormalities appear early in LC, and that these abnormalities can be detected by MIBG myocardial scintigraphy and analysis of heart-rate variability. We consider these methods to be clinically useful for the quantitative detection of hyperdynamic circulation of liver cirrhosis. PMID- 12753147 TI - Scirrhous changes in dysplastic nodules do not indicate high-grade status. AB - BACKGROUND AND AIMS: Dysplastic nodules (DN) may be divided into high-grade and low-grade, and the former has been known as a precancerous or borderline lesion. Recently many morphological characteristics concerning these types of DN have been reported. In the present study we attempted to evaluate the scirrhous change in DN as an indicative feature of high-grade DN, based on the morphological and cell-kinetic analyses using immunohistochemical stains for Ki-67. METHODS: We reviewed 35 livers with DN and selected 15 DN with scirrhous change. We stained DN-bearing sections of each case with hematoxylin and eosin, trichrome, reticulin and Perls' stain. We tried to subclassify and characterize the scirrhous change according to the fibrosis pattern. We also stained with Ki-67 immunohistochemically to assess the proliferative activity of DN with scirrhous change. RESULTS: We found two types of scirrhous change, that is, pericellular and stellate. The pericellular type was related to the Mallory body-forming cholestatic degeneration, whereas the stellate type was associated with extensive portal fibrosis probably induced by ischemic damage. Among DN with scirrhous change, high-grade DN comprised five nodules (33%) and there were 10 (67%) low grade nodules. There was no significant relationship between the presence or the types of scirrhous change and the grade of DN. The significant differences of Ki 67 labeling indices between types of scirrhous change were not shown in this study. We also could not find the differences between Ki-67 labeling indices of scirrhous DN (high and low grades) and those of surrounding regenerative nodules. CONCLUSIONS: This evidence indicated that the scirrhous change in DN was not a specific feature of high-grade DN. We also found that scirrhous DN have two morphological varieties that may represent biologically different processes, that is, pericellular scirrhous type and stellate scirrhous type. PMID- 12753148 TI - Tumor progression in hepatocellular carcinoma: relationship with tumor stroma and parenchymal disease. AB - BACKGROUND: Encapsulation in hepatocellular carcinoma is associated with decreased invasiveness and improved survival in several series. Although active fibrogenesis by myofibroblasts has been demonstrated in the capsule, it is unclear if the capsule results from a general increase in peritumoral fibrosis, or an inherently less invasive tumor phenotype. The relationship between collagen deposition within tumor stroma, presence of cirrhosis and invasiveness also needs clarification. METHODS: We performed immunohistochemistry for collagens I, III, IV and VI on sections of encapsulated and non-encapsulated hepatocellular carcinoma, arising in cirrhotic and non-cirrhotic livers. Staining was graded semi-quantitatively in tumor stromal elements and adjacent parenchymal sinusoids. The relationship of this staining with encapsulation, cirrhosis, and vascular invasion was analyzed. RESULTS: Formation of a discrete capsular layer was associated with reduced vascular invasion, but not with a pervasive increase in peritumoral fibrosis. Increased collagen I content of tumor stroma and adjacent parenchymal sinusoids was associated with non-encapsulated tumors and vascular invasion. The presence of cirrhosis had little effect on capsule composition. CONCLUSIONS: Encapsulation of hepatocellular carcinoma reflects reduced invasiveness, rather than increased peritumoral collagen synthesis, which may instead enhance invasion. Increased intratumoral collagen I protein is also associated with increased tumor invasiveness. Pre-existing cirrhosis has little effect on tumor progression, possibly because the characteristics of cirrhosis are overwhelmed by tumor-induced changes in the adjacent parenchyma. PMID- 12753149 TI - Gross appearance of hepatocellular carcinoma reflects E-cadherin expression and risk of early recurrence after surgical treatment. AB - BACKGROUND AND AIMS: Correlation between the gross classification of hepatocellular carcinoma (HCC) and its vascular invasion or intrahepatic metastasis has been reported previously. Because E-cadherin-mediated epithelial cell-to-cell adhesion is thought to suppress cancer cell invasion, the present study was performed to analyze the correlation between E-cadherin expression and the gross classification of HCC. METHODS: Thirty-six resected solitary HCC <6 cm in diameter were each classified as single nodular type (type 1), single nodular with extranodular growth type (type 2) or contiguous multinodular type (type 3), and the clinicopathological and prognostic differences between type 1 HCC and the other types were analyzed. The expression of E-cadherin in each tumor was examined by immunoblotting and immunohistochemical analysis. RESULTS: Vascular invasion and microscopic intrahepatic metastasis were observed more frequently in types 2 and 3 (61%) than in type 1 (13%) HCC. Immunoblot analysis indicated that the relative level of E-cadherin expression in cancerous tissue was significantly lower in type 2 and 3 (0.75 +/- 0.49) than in type 1 (1.46 +/- 0.79) HCC. Immunohistochemical examination revealed decreased and partially absent E cadherin expression in the tumorous area of type 2 and 3 HCC. The recurrence-free survival rate was higher for patients with type 1 HCC than for those with the other types. CONCLUSIONS: Types 2 and 3 HCC have marked metastatic and invasive potential and reduced expression of E-cadherin, predicting a high risk of recurrence after surgical treatment. PMID- 12753150 TI - Tolerance to famotidine and ranitidine treatment after 14 days of administration in healthy subjects without Helicobacter pylori infection. AB - BACKGROUND AND AIM: The attenuated antisecretory activity observed during continuous administration of ranitidine has been described as tolerance. However, it remains unclear whether a similar phenomenon occurs with other histamine H2 receptor antagonists (H2RA). We investigated whether tolerance to famotidine, a stronger H2RA than ranitidine, occurs during long-term administration. METHODS: Seven healthy male Japanese subjects without Helicobacter pylori infection participated in a randomized cross-over study in which ranitidine and famotidine were administered for 14 days with a 4-week wash-out period. We performed 24-h intragastric pH monitoring on the first and 14th days of administration of each drug, and measured serum gastrin and plasma drug concentrations on the first, seventh and 14th days. RESULTS: The acid-inhibiting activity of ranitidine and famotidine declined during continuous administration. In particular, the potent nocturnal pH-increasing effect of the H2RA, which was observed on day 1, declined on day 14. Serum gastrin concentrations on day 14 were significantly lower than those on day 7, although plasma drug concentrations remained unchanged. CONCLUSION: Tolerance to famotidine occurs during continuous administration for 14 days, as previously shown in ranitidine studies. PMID- 12753151 TI - Loss of heterozygosity on chromosome 17p predicts neoplastic progression in Barrett's esophagus. AB - BACKGROUND AND AIM: Endoscopic surveillance for adenocarcinoma in patients with Barrett's esophagus is costly, with one cancer detected every 48-441 patient years of follow up. Genetic abnormalities, including loss of heterozygosity at sites of tumor suppressor genes, have been detected in malignant and premalignant Barrett's esophagus. The aim of this prospective study was to determine if loss of heterozygosity analysis could identify patients with Barrett's esophagus at greatest risk of adenocarcinoma, for whom endoscopic surveillance is most appropriate. METHODS: Loss of heterozygosity analysis was performed on endoscopic biopsies from 48 patients as part of a Barrett's surveillance program using 14 microsatellite markers shown previously to detect loss of heterozygosity in more than 30% of esophageal adenocarcinomas. Patients were followed up endoscopically for a median of 5 years. RESULTS: Loss of heterozygosity was detected in nine patients. Three patients with loss of heterozygosity on chromosome 5q or 9p did not progress beyond metaplasia. Loss of heterozygosity at 17p11.1-p13 was detected in six patients, all of whom demonstrated dysplasia and/or carcinoma during follow up (four low-grade dysplasia, one high-grade dysplasia and one adenocarcinoma). CONCLUSION: Loss of heterozygosity at 17p11.1-p13 on chromosome 17p identifies patients with Barrett's esophagus at risk of neoplastic progression and can supplement histology in determining the frequency of endoscopy during surveillance. PMID- 12753152 TI - Differential, stage-dependent expression of Hsp70, Hsp110 and Bcl-2 in colorectal cancer. AB - BACKGROUND: The presence of hypoxic cells in solid tumors has been suggested to contribute to the malignant progression of various tumors. Recently, we reported an activation of heat shock transcription factor (HSF) and expression of heat shock proteins (Hsp) in murine tumor cells by hypoxia. METHODS: To search for a possible role of Hsp in the malignant progression of human tumors, we analyzed the expression profiles of Hsp family proteins in weakly and highly metastatic human colorectal cancer (CRC) cell lines. We also analyzed the expression profiles of Bcl-2 family proteins because the altered expression of these proteins has been demonstrated in various solid tumors. RESULTS: In the present paper we showed among various Hsp and Bcl-2 family proteins that the expression of Hsp70 and Hsp110 was elevated in highly metastatic CX-1 and HT-29 cells, while the expression of Bcl-2 was elevated in weakly metastatic MIP-101 and Clone A cells. Subsequent immunohistochemical analysis of 81 primary human CRC tissues demonstrated that the expression of Hsp70 and Hsp110 was highly correlated with the advanced clinical stages and positive lymph node involvement. The expression of Bcl-2, in contrast, was correlated to the early clinical stage and negative lymphovascular invasion. CONCLUSION: Taken together, our study demonstrated for the first time a differential, stage-dependent expression of Hsp70, Hsp110 and Bcl-2 in CRC. We suggest that the molecular mechanisms underlying the differential expression of Hsp and Bcl-2 family members deserves a more rigorous future study, the results of which might offer novel modes of rationale and strategy to predict and manipulate the malignant progression of colorectal cancers. PMID- 12753153 TI - Flat colonic adenomas in Malaysia: fact or fancy? AB - BACKGROUND: Some two-thirds of colorectal carcinomas arise from adenomatous polyps, and as such, screening by colonoscopy and polyp removal should significantly reduce colorectal cancer. This has not been the case, as evidenced by recent studies, which revealed that endoscopy failed to prevent up to 50% of all subsequent carcinomas. Flat or depressed adenomas, frequently reported from Japan but rarely elsewhere, might explain the 'missed carcinomas.' Detection of flat adenomas has not been previously reported from Malaysia. METHODS: In the present prospective study, 426 consecutive patients underwent colonoscopic examination between March 1997 and January 2000, for a variety of bowel symptoms. The examinations were performed by an experienced endoscopist using a standard colonoscope and methylene blue dye spraying technique. Macroscopically, flat adenomas were defined using the criteria proposed by Sawada. RESULTS: Twenty-nine adenomas were identified in 12 patients, of which 15 were polypoid and 14 were flat, with no depressed lesions. Eight polypoidal lesions and all the flat adenomas contained mild or moderate areas of epithelial dysplasia. Seven severely dysplastic polyps were identified. One Duke's A polypoidal cancer and two advanced carcinomas were also found. All the severely dysplastic lesions and Duke's A carcinomas were found in polyps greater than 10 mm in mean size. The flat adenomas were all less than 5 mm in size. CONCLUSIONS: A significant proportion of colonic adenomas in Malaysian patients appear as small flat lesions, which could easily be missed during endoscopy. Increased recognition and treatment of flat adenomas among colonoscopists is warranted. PMID- 12753154 TI - Alterations in the DNA binding activity of transcriptional factors activator protein-1, Sp1, and hepatocyte nuclear factor-1 in rat jejunum during starvation and refeeding. AB - BACKGROUND: The molecular processes leading to mucosal atrophy, regrowth, and functional changes with starvation and refeeding are largely unknown. There are many transcriptional factors that might be related to mucosal atrophy and proliferation. In contrast, we previously reported that H+/peptide transporter and aminopeptidase N messenger RNA in the intestinal mucosa were upregulated during starvation. Therefore, we selected and studied three transcriptional factors: activator protein (AP)-1, Sp1, and hepatocyte nuclear factor (HNF)-1, which not only play important roles for enterocytes proliferation, but also exist in promoter lesions of the brush border enzymes and peptide transporter. METHODS: In the present study, we performed electrophoretic mobility shift assays employing AP-1, Sp1, and HNF-1, and evaluated the changes in the DNA binding activities in rat jejunum during starvation and refeeding. RESULTS: Two days after starvation, the Sp1 binding activity was significantly decreased to 61.8% as compared with the control level, whereas AP-1 was 121.4% and HNF-1 was 77.5%. Two hours after refeeding, the AP-1 activity was significantly increased to 175.0% as compared with the control level, and the HNF-1 activity was significantly increased to 180.2%. In contrast, the decreased SP1 level did not recover until 24 h after refeeding. CONCLUSIONS: The DNA binding activities of these three transcriptional factors were significantly changed in the rat jejunum during starvation and refeeding. Our results provide insight into the molecular mechanisms of the transcriptional regulations associated with mucosal atrophy, regrowth, and functional changes of the jejunal epithelium in response to starvation and refeeding. PMID- 12753155 TI - Novel human and mouse genes encoding a shank-interacting protein and its upregulation in gastric fundus of W/WV mouse. AB - BACKGROUND AND AIMS: A division of labor exists between different classes of interstitial cells of Cajal (ICC) in the gastrointestinal tract. In the stomach and small intestine, ICC at the level of the myenteric plexus (IC-MY) act as slow wave pacemaker cells, whereas intramuscular ICC (IC-IM) in the stomach act as intermediaries in enteric motor neurotransmission. The muscle layers of the gastric fundus do not have IC-MY, therefore electric slow waves are not generated. Intramuscular ICC are absent in the gastric fundus of W/WV mutant mice, and excitatory and inhibitory motor nerve responses are reduced in these tissues. The absence of IC-IM in W/WV mutants in the fundus provides a unique opportunity to study the molecular changes that are associated with the loss of these cells. METHODS: The tissue gene expression of wild-type and W/WV mice from gastric fundus was assayed using a murine microarray chip analysis displaying a total of 8734 elements. RESULTS: Twenty-one queries were differentially expressed in wild-type and W/WV mice. One candidate gene, encoding a novel protein homologous to rat Shank-interacting protein (Sharpin) was significantly upregulated in fed and starved W/WV mice. The full-length clone of the murine gene and its human counterpart were isolated and designated as Shank-interacting protein-like 1 (SIPL1). Human SIPL1 complementary DNA encodes a protein of 345 amino acids. This gene was localized to chromosome 8. SIPL1 was abundantly expressed in human stomach and small intestine, and scarcely expressed in cecum and rectum. CONCLUSIONS: Gene analysis showed that SIPL1 differentially express in the gastric fundus of normal and W/WV mice. The upregulation of SIPL1 in the fundus of W/WV mice, and expression in the upper gastrointestinal tract suggest that the SIPL1 gene could be associated with ICC function in mice and humans. PMID- 12753156 TI - Nutritional benefits of enteral alanyl-glutamine supplementation on rat small intestinal damage induced by cyclophosphamide. AB - BACKGROUND: Glutamine is the principal fuel used by the small intestine. Although the parental administration of glutamine promotes intestinal mucosal growth, it is controversial whether enteral glutamine is effective against small intestinal damage caused by chemotherapy. To further evaluate the benefits of enteral supplementation, peptide and amino acid transporter functions must be considered. METHOD: Rats were given cyclophosphamide (CPM) intraperitoneally (300 mg/kg). Expression of the amino acid transporter, B0 and peptide transporter (PepT1) in the jejunal mucosa was initially examined by northern blot analysis. Rats received a bolus oral supplement of an alanine (1.22 g/kg/day) plus glutamine (2.0 g/kg/day) mixture, alanyl-glutamine (2.972 g/kg/day) or saline as a control, for 7 days after CPM administration. RESULTS: Levels of B0 mRNA remained unchanged at both 3 and 7 days after CPM administration. Conversely, PepT1 mRNA increased significantly after CPM administration, and reached 200% of the initial level 7 days later. In rats given alanyl-glutamine, the mucosal wet weight and protein content increased significantly with increasing villus height at 3 and 7 days, compared with the alanine plus glutamine mixture. The plasma glutamine concentration in the alanyl-glutamine group, but not the alanine plus glutamine mixture group, increased significantly compared with that in the saline group. CONCLUSION: Enteral supplementation with an alanyl-glutamine but not alanine plus glutamine mixture prevents intestinal damage, as demonstrated by increased peptide transport expression and an elevated plasma glutamine concentration after CPM administration. PMID- 12753157 TI - Delta (13)C-urea breath test value is a useful indicator for Helicobacter pylori eradication in patients with functional dyspepsia. AB - BACKGROUND: Eradication of Helicobacter pylori is not routinely recommended for the symptomatic relief and the prevention of gastric cancer in patients with functional dyspepsia. The present study investigated a useful indicator of H. pylori eradication in such patients by determining the optimal cutoff value of a 13C-urea breath test (UBT). METHODS: One hundred dyspeptic patients participated in the study. Dyspepsia was scored, and a 13C-UBT administered. A level of delta 13C-UBT of>4 per thousand was diagnosed as H. pylori-positive. After the stomach was endoscopically sprayed with phenol red, biopsy specimens were taken from the antrum, body and cardia of the stomach for the assessment of H. pylori density, and activity (neutrophil infiltration) and degree (lymphocyte infiltration) of gastritis. RESULTS: Correlation between delta 13C-UBT and dyspepsia score was not found. Delta 13C-UBT significantly correlated with H. pylori density score in the total stomach (r = 0.53, P < 0.0001), neutrophil (r = 0.34, P = 0.0005) and lymphocyte score (r = 0.69, P < 0.0001). Twenty-six of the 100 subjects had a neutrophil score of >or=4, lymphocyte score of >or=4, and H. pylori score of >or=4. Their 95% confidence interval of mean was 58.2 per thousand, which reflects moderate to marked acute and chronic gastritis, and dense H. pylori colonization. CONCLUSIONS: The 13C-UBT is a reliable semiquantitative test to assess H. pylori density and the activity and degree of gastritis. It is proposed that H. pylori eradication therapy might be beneficial for patients with functional dyspepsia with a delta 13C-UBT of >58.2 per thousand. PMID- 12753159 TI - Hepatobiliary and pancreatic: a boy with anorexia and an abnormal gallbladder. PMID- 12753158 TI - Evaluation of Helicobacter pylori stool antigen test before and after eradication therapy. AB - BACKGROUND AND AIM: The Helicobacter pylori stool antigen (HpSA) test is useful for initial diagnosis of H. pylori infection, but there is disagreement regarding its diagnostic accuracy after eradication therapy. The aim of the present study was to evaluate the diagnostic accuracy of the HpSA test before and after eradication therapy. METHODS: One hundred and thirty-six patients underwent upper gastrointestinal endoscopy with biopsies for the diagnosis of H. pylori infection using culture, histology and the rapid urease test. Fifty-four H. pylori-positive patients were treated with 1-week triple therapy. Six to 10 weeks after the end of therapy, the patients underwent re-endoscopy and received the same biopsy based methods. In addition, the 13C-urea breath test was performed. The HpSA test was performed before and 6-10 weeks after the end of therapy. In 23 patients, the HpSA test was also performed at the end of therapy. RESULTS: Before therapy, the sensitivity and specificity of the HpSA test was 98.3% (95% confidence interval (CI): 95.9-100%) and 95.0% (95% CI: 75.1-99.9%), respectively. At the end of therapy, the HpSA tests were all negative both for eradication and non eradication patients. The sensitivity and specificity of the HpSA test after eradication therapy were 90% (95% CI: 55.5-99.7%) and 97.7% (95% CI: 93.3-100%), respectively. CONCLUSIONS: The HpSA test is a useful method for the diagnosis of H. pylori infection before and after eradication therapy. PMID- 12753160 TI - Gastrointestinal: cap polyposis. PMID- 12753162 TI - Primary small cell carcinoma of the stomach. AB - We report on an 80-year-old man with primary gastric small cell carcinoma (SmCC). He was admitted to hospital with hematemesis. An upper gastrointestinal examination revealed an irregularly ulcerated tumor, 60 mm in diameter, on the lesser curvature of the stomach body extending to the cardia. An endoscopic biopsy revealed a solid proliferation of intermediate-sized tumor cells with hyperchromatic nuclei and scanty cytoplasm. Immunohistochemically, the neoplastic cells were positive for neuron-specific enolase and chromogranin A, but negative for carcinoembryonic antigen. No tumor was detected on examination of the chest. Therefore, primary gastric SmCC was diagnosed preoperatively. To date, only 38 cases of primary gastric SmCC, including our case, have been reported. By using endoscopic biopsy, approximately two-thirds of cases have been diagnosed incorrectly. In the reported cases of gastric SmCC, the endoscopic findings frequently indicated a submucosal tumor. Gastric SmCC is clinically aggressive and has an extremely poor prognosis, even when discovered at an early stage. Most patients with gastric SmCC die within 1 year of diagnosis. Although a standard treatment for gastric SmCC has not been established, intensive chemotherapy should be considered to promote long-term survival. We believe that careful examination, including immunohistochemical investigation, is necessary for determining the therapeutic strategy whenever gastric SmCC is suspected during endoscopy. PMID- 12753163 TI - Colonic carcinosarcoma. PMID- 12753164 TI - Severe intestinal involvement in Wegener's granulomatosis. PMID- 12753165 TI - Children's age affects hepatitis B vaccine response. PMID- 12753167 TI - Under the microscope: doctors, lawyers, and melanocytic neoplasms. AB - Misdiagnosed melanoma remains one of the most common causes of lawsuits in histopathology. Reasons for this are discussed in conjunction with relevant literature, common errors in diagnosis, and strategies to avoid them. PMID- 12753168 TI - Cutaneous myoepithelial neoplasms: clinicopathologic and immunohistochemical study of 20 cases suggesting a continuous spectrum ranging from benign mixed tumor of the skin to cutaneous myoepithelioma and myoepithelial carcinoma. AB - BACKGROUND: Myoepithelial neoplasms, both benign and malignant, are rare but well established clinicopathologic entities in the salivary glands, the breast, and the lung. Despite similarities between cutaneous sweat glands and glandular structures in the above-mentioned organs as well as the presence of regular myoepithelial cells around cutaneous eccrine/apocrine glands, the concept of cutaneous myoepithelial neoplasms is still debatable and not commonly accepted. METHODS: Twenty cutaneous myoepithelial neoplasms have been studied histologically and immunohistochemically. RESULTS: Nine neoplasms showed features of benign mixed tumor of the skin (chondroid syringoma) (five females and four males, age range 19-65 years, all cases arose in the head and neck region). Two cases represented the eccrine and seven the apocrine subtype. Interestingly, in three cases of the apocrine subtype, solid areas composed predominantly of myoepithelial cells were detected; these neoplasms were designated as benign mixed tumors with prominent myoepithelial cells. Nine cutaneous neoplasms were composed of spindled, epithelioid, and plasmocytoid cells without ductal differentiation and immunohistochemically stained variably positive for vimentin, epithelial and myogenic markers, S-100 protein, calponin, and glial fibrillary acidic protein (four females and five males, age range 3-71 years, four cases arose in the head and neck region and one case each on the finger, the thigh, the lower leg, the foot, and the breast, respectively); these neoplasms were designated as cutaneous myoepitheliomas. Two morphologically malignant neoplasms with cytologic and immunohistochemical features of myoepithelial cells arose on the face of a 70-year-old female and a 79-year-old male patient; these neoplasms were designated as malignant cutaneous myoepitheliomas (cutaneous myoepithelial carcinomas). CONCLUSIONS: The study suggests a continuous spectrum of cutaneous myoepithelial neoplasms ranging from benign mixed tumor of the skin to cutaneous myoepithelioma and cutaneous myoepithelial carcinoma. Further studies with extended follow-up information are necessary to establish prognostic factors. PMID- 12753169 TI - Tick mouth parts occlusive vasculopathy: a localized cryoglobulinemic vasculitic response. AB - BACKGROUND: The histologic hallmark of most arthropod bite reactions is a deep, wedge-shaped perivascular and interstitial infiltrate comprising lymphocytes, neutrophils, and eosinophils. METHODS: We present a case series of six patients in whom tick bite reactions, when examined microscopically, were found to mimic mixed cryoglobulinemic vasculitis. RESULTS: Though different in histology, clinically these lesions were indistinguishable from typical tick bite reactions. CONCLUSION: As five of our six biopsy specimens were found to still harbor retained tick parts, it is possible that the actual retention of tick parts was involved in evoking this localized cryoprecipitate reaction. PMID- 12753170 TI - Ofuji's disease with follicular mucinosis and its differential diagnosis from alopecia mucinosa. AB - OBJECTIVES: Ofuji's disease (OD) or eosinophilic pustular folliculitis and human immunodeficiency virus-associated eosinophilic folliculitis (HIV-EF) both show eosinophil-rich folliculocentric infiltrates, and it is not clear whether they are distinguishable pathologically. Follicular mucinosis (FM) has been observed in lesions of EF; such cases need to be differentiated from alopecia mucinosa (AM). METHODS: We compared various pathologic features in 13 cases of OD, 10 cases of non-OD-typed papular EF (seven HIV-positive and three HIV-negative), and five cases of AM. RESULTS: All cases of EF showed eosinophilic infiltrates affecting mainly the isthmus or/and sebaceous gland. Eosinophil-rich pilosebaceous pustules or/and microabscesses were noted in 69% of the biopsy specimens of OD, 73% of papular EF, 71% of HIV-EF, and 0% of AM. Mucin deposits, often abundant, were found in sebaceous lobules or/and isthmus in 41% of OD and 100% of AM. Compared with AM, OD with FM tended to show more numerous eosinophils, less abundant mucin, and most significantly, eosinophilic infundibular pustule or sebaceous microabscess. CONCLUSION: Our study suggests that OD and HIV-EF are indistinguishable pathologically and the diagnosis requires clinical correlation. FM was not uncommonly seen in lesions of OD. OD with FM may be difficult to differentiate from FM, especially in cases presenting with non-annular or non-pustular lesions, but the diagnosis might be facilitated by finding eosinophil-rich pustule, microabscess, or infiltrate in pilosebaceous units microscopically. PMID- 12753171 TI - Immunofluorescence findings in granuloma faciale: report of two cases. AB - BACKGROUND: Immunofluorescence findings in granuloma faciale have been infrequently described. Reported findings include granular IgA, IgG, IgM, and C3 deposits in the dermoepidermal junction, in blood vessel walls, and on connective tissue fibers; IgG in the basement membrane zone, and IgG around blood vessels. OBSERVATIONS: We report two cases of granuloma faciale in which biopsy specimens were sent for routine pathology as well as immunofluorescence. Additional work-up included serologic testing for markers of lupus erythematosus (LE). Immunofluorescence in both cases revealed granular IgA, IgG, IgM, C3, and C5-9 deposits along the dermoepidermal junction only. Blood vessel walls and connective tissue fibers were spared. Serologic studies for markers of LE were negative. CONCLUSION: Immunofluorescence findings were essentially identical to those which would be expected in cutaneous LE; however, the combination of negative serologic studies for LE, histopathologic findings typical of granuloma faciale, and clinical findings most consistent with granuloma faciale allowed the diagnosis of granuloma faciale to be rendered with certainty. Interpretation of results of direct immunofluorescence of skin should be correlated with clinical presentation, histopathological findings and other laboratory results in order to render final diagnosis of a given patient. PMID- 12753172 TI - Expression of neurotrophin receptor Trk-C in nevi and melanomas. AB - BACKGROUND: Neurotrophins (NTs) are growth factors for neurons and other neural crest-derived cells. Their functions are mediated by 75-kDa low-affinity glycoprotein receptor (p75) NT receptor and a family of tyrosine kinase receptors (Trks) that includes Trk-A, -B, and -C. Signal transduction through the Trk receptors has been shown to regulate growth and apoptosis of tumors of neuronal origin. In addition, Trk oncogenes have been shown to be rearranged in some non neuronal neoplasms. Recently, immunoexpression of NT-3 has been shown to be significantly higher in melanomas than in banal nevi on cryostat tissue. METHODS: Since the biologic function of NT-3 is mediated primarily through Trk-C, we investigated Trk-C immunoexpression on paraffin sections of 10 compound nevi and 63 melanomas. RESULTS: The expression of Trk-C was relatively low in compound nevi (30%). Trk-C expression was overall 62% in melanomas of various stages. Our data show that the expression of Trk-C increased as melanoma progressed from in situ (58%) to papillary dermal invasion (91%), and then declined in deeper (57%) and metastatic melanomas (31%). CONCLUSION: These findings suggest a possible role of Trk-C in the progression of early stages of melanoma. PMID- 12753173 TI - Metastasis of cutaneous malignant melanoma to angiolipoma: the tumor-to-tumor metastasis phenomenon. AB - BACKGROUND: Although the phenomenon of tumor-to tumor metastasis is not rare, it has been reported in only eight cases with malignant melanoma as the primary tumor. METHODS: This case describes a patient with cutaneous malignant melanoma that metastasized to an angiolipoma. To our knowledge this is the second case of a malignant melanoma metastasizing to another primary cutaneous tumor and the first to do so in a lipoma. CONCLUSION: In this report we present the clinical and histopathologic features of this special case and review the relevant literature. PMID- 12753174 TI - Brainstem involvement by mycosis fungoides in a patient with large-cell transformation: a case report and review of literature. AB - BACKGROUND: Central nervous system (CNS) involvement by mycosis fungoides (MF) is rare. As compared to meningeal involvement, intraparenchymal spread is especially rare. It is usually seen in advanced disease in conjunction with lymph node or visceral involvement. However, CNS involvement in the absence of progressive skin lesions or other extracutaneous involvement has been reported rarely in patients with transformed MF. METHOD: Case report and review of literature. RESULTS: A 71 year-old female with long-standing MF developed lymphomatous CNS involvement 10 years after the diagnosis of tumor stage MF. At this time, the patient presented with a transient episode of garbled speech followed by generalized weakness. Computerized tomography scan (CT scan) and magnetic resonance imaging scan (MRI scan) of the head revealed a subcortical lesion in the left temporo-frontal lobe. Cerebrospinal fluid (CSF) examination showed atypical T cells, and brain biopsy confirmed parenchymal involvement by T-cell lymphoma. Meanwhile, a biopsy of a skin lesion showed large-cell transformation. No lymph node or other systemic involvement was noted at this time, and the patient was treated with chemotherapy. Twelve months later, the patient developed recurrent CNS lymphoma with multiple organ involvement and expired soon thereafter. CONCLUSIONS: This case illustrates the importance of awareness of the possibility of CNS involvement by MF, especially in transformed MF. CNS involvement may be the only site of extracutaneous involvement in patients with transformed MF, and mental status changes warrant CNS surveillance for this disease. PMID- 12753175 TI - Granular parakeratosis: a case report and literature review. AB - BACKGROUND: Granular parakeratosis is suspected to result from an error in epidermal differentiation, leading to variably pruritic, hyperpigmented-to erythematous patches and plaques. Characteristic histopathologic features include a thickened stratum corneum, compact parakeratosis with retention of keratohyalin granules, vascular proliferation, and ectasia. The pathogenesis of this entity is uncertain. METHODS: We present a case of axillary granular parakeratosis and review the literature. RESULTS: The lesion showed a thickened stratum corneum with compact parakeratosis, slight epidermal hyperplasia, and a sparse perivascular lymphohistiocytic infiltrate. Keratohyalin granules were diffusely present within the parakeratotic stratum corneum, and the retained granular layer showed focal vacuolization. CONCLUSIONS: Granular parakeratosis is a rare form of parakeratosis most often seen in the axilla, although other intertriginous areas may be affected. Unique histopathologic findings allow for a specific diagnosis to be made. Although an irritant contact reaction appears causative, mechanical irritation may also play a role in inducing these skin changes. PMID- 12753176 TI - Atypical fibroxanthoma with prominent sclerosis. AB - BACKGROUND: Malignant cutaneous spindle cell lesions with marked sclerosis are uncommon. Only a few cases of cutaneous leiomyosarcoma and dermatofibrosarcoma protuberans with sclerosis have been published. METHODS: We report a case of atypical fibroxanthoma (AFX) with prominent sclerosis and hyalinization occurring on the scalp of an 81-year-old male. RESULTS: Histopathologic examination revealed an exophytic, well-delineated, focally ulcerated tumor arising in sun damaged skin. The lesion was composed of atypical spindle cells arranged in a fascicled and vaguely storiform pattern. Occasional multinucleated giant cells were present. The tumor cells were strongly positive for CD99 (O13), vimentin, and smooth muscle actin, and focally positive for CD68. There was striking sclerosis with hyalinization throughout the lesion. CONCLUSIONS: Rarely, AFX may exhibit marked sclerosis with areas of complete replacement of tumor by hyalinized collagen. In a small biopsy, such hyalinization may be a diagnostic pitfall leading to an erroneous diagnosis. PMID- 12753177 TI - An unusual dematiaceous fungal infection of the skin caused by Fonsecaea pedrosoi: a case report and review of the literature. AB - BACKGROUND: A case of an unusual dematiaceous fungal infection of the skin in a 43-year-old man with diabetes mellitus treated with steroids for reactive airway disease is presented. He developed chromoblastomycosis in the left wrist and was treated with antifungals and multiple surgical excisions. RESULTS: Histologic examination of the excised tissue revealed widespread suppurative granulomatous inflammation in the dermis and subcutaneous tissue. Thick-walled internally septated brown fungal cells were found both inside multinucleated giant cells and extracellularly. Non-to-lightly pigmented septate hyphal elements, however, were also identified with special stains and, in retrospect, on one of the routinely stained sections. In culture, the organism was reported to initially grow as soft white colonies that soon turned to black and velvety. CONCLUSIONS: The two unusual features of this case include the controversial report of the organism's initial growth in culture as soft white colonies and the presence of hyphal elements in addition to the sclerotic bodies in the dermis and subcutaneous tissue. This has not been reported before in human cases of dermal infection by Fonsecaea pedrosoi. PMID- 12753178 TI - Multiple human papillomavirus DNA identified in verruciform xanthoma by nested polymerase chain reaction with degenerate consensus primers. PMID- 12753181 TI - Small non-coding RNAs, co-ordinators of adaptation processes in Escherichia coli: the RpoS paradigm. AB - Adaptation to the changing environment requires both the integration of external signals and the co-ordination of internal responses. Around 50 non-coding small RNAs (sRNAs) have been described in Escherichia coli; the levels of many of these vary with changing environmental conditions. This suggests that they play a role in cell adaptation. In this review, we use the regulation of RpoS (sigma38) translation as a paradigm of sRNA-mediated response to environmental conditions; rpoS is currently the only known gene regulated post-transcriptionally by at least three sRNAs. DsrA and RprA stimulate RpoS translation in response to low temperature and cell surface stress, respectively, whereas OxyS represses RpoS translation in response to oxidative shock. However, in addition to regulating RpoS translation, DsrA represses the translation of HNS (a global regulator of gene expression), whereas OxyS represses the translation of FhlA (a transcriptional activator), allowing the cell to co-ordinate different pathways involved in cell adaptation. Environmental cues affect the synthesis and stability of specific sRNAs, resulting in specific sRNA-dependent translational control. PMID- 12753182 TI - Region 4 of sigma as a target for transcription regulation. AB - Bacterial sigma factors play a key role in promoter recognition, making direct contact with conserved promoter elements. Most sigma factors belong to the sigma70 family, named for the primary sigma factor in Escherichia coli. Members of the sigma70 family typically share four conserved regions and, here, we focus on region 4, which is directly involved in promoter recognition and serves as a target for a variety of regulators of transcription initiation. We review recent advances in the understanding of the mechanism of action of regulators that target region 4 of sigma. PMID- 12753183 TI - Identification and macrophage-activating activity of glycolipids released from intracellular Mycobacterium bovis BCG. AB - Intracellular mycobacteria release cell wall glycolipids into the endosomal network of infected macrophages. Here, we characterize the glycolipids of Mycobacterium bovis BCG (BCG) that are released into murine bone marrow-derived macrophages (BMMO). Intracellularly released mycobacterial lipids were harvested from BMMO that had been infected with 14C-labelled BCG. Released BCG lipids were resolved by thin-layer chromatography, and they migrated similarly to phosphatidylinositol dimannosides (PIM2), mono- and diphosphatidylglycerol, phosphatidylethanolamine, trehalose mono- and dimycolates and the phenolic glycolipid, mycoside B. Culture-derived BCG lipids that co-migrated with the intracellularly released lipids were purified and identified by electrospray ionization mass spectrometry. When delivered on polystyrene microspheres, fluorescently tagged BCG lipids were also released into the BMMO, in a manner similar to release from viable or heat-killed BCG bacilli. To determine whether the released lipids elicited macrophage responses, BCG lipid-coated microspheres were delivered to interferon gamma-primed macrophages (BMMO or thioglycollate elicited peritoneal macrophages), and reactive nitrogen intermediates as well as tumour necrosis factor-alpha and monocyte chemoattractant protein-1 production were induced. When fractionated BCG lipids were delivered on the microspheres, PIM2 species reproduced the macrophage-activating activity of total BCG lipids. These results demonstrate that intracellular mycobacteria release a heterogeneous mix of lipids, some of which elicit the production of proinflammatory cytokines from macrophages that could potentially contribute to the granulomatous response in tuberculous diseases. PMID- 12753184 TI - Identification of a 5-nucleotide sequence that controls expression of the ica locus in Staphylococcus aureus and characterization of the DNA-binding properties of IcaR. AB - Biofilm formation is an important aspect of the pathogenesis of staphylococcal infections. A beta-1,6-linked N-acetyl glucosamine polysaccharide is critical to biofilm elaboration and is synthesized by proteins encoded by the intercellular adhesion (ica) locus. These studies were undertaken to characterize the mechanism by which transcription of the ica locus in S. aureus is regulated using isogenic S. aureus MN8 and MN8 mucoid (MN8m) strains, the latter of which constitutively overproduces biofilm. Transformation of the ica locus from MN8m to the ica knock out mutants of two strains, MN8 and NCTC 10833, conferred a strong biofilm producing phenotype. Sequence analysis revealed a 5-nucleotide deletion within the promoter region of the ica locus in MN8m compared with the sequence in the wild-type locus. Deletion or substitution of these 5 nucleotides within the wild type ica locus augmented transcription of the ica locus and induced the strong biofilm-producing phenotype. Gel shift analysis demonstrated that a protein(s) within cell-free lysates from strain MN8 bind(s) specifically to oligonucleotides representative of the wild-type ica promoter sequence and that this binding is greatly diminished by the deletion or substitution of the 5 nucleotides. DNase I footprint analysis revealed that purified IcaR, thought to be a regulator of ica transcription, also binds to the ica promoter sequence just upstream of the ica start codon, but its affinity for the ica promoter is unaffected by deletion of the 5-nucleotide motif. These findings identify a 5-nucleotide motif within the ica promoter region that has a functional role in transcriptional regulation of the ica locus that is independent of IcaR, and also show that IcaR binds to the promoter region of the S. aureus ica locus. PMID- 12753185 TI - Sequence-specific recognition but position-dependent cleavage of two distinct telomeres by the Borrelia burgdorferi telomere resolvase, ResT. AB - An unusual feature of bacteria in the genus Borrelia (causative agents of Lyme disease and relapsing fever) is a segmented genome consisting of multiple linear DNA molecules with covalently closed hairpin ends, known as telomeres. The hairpin telomeres are generated by a DNA breakage and reunion process (telomere resolution) promoted by ResT, an enzyme using an active site related to that of tyrosine recombinases and type IB topoisomerases. In this study, we define the minimal sequence requirements for a functional telomere and identify specific basepairs that appear to be important for telomere resolution. In addition, we show that the two naturally occurring and distinct telomere spacings found in B. burgdorferi can both be efficiently processed by ResT. This flexibility for substrate utilization by ResT supports the argument for a single telomere resolvase in Borrelia. Furthermore, although telomere recognition requires sequence specificity in part of the substrate, DNA cleavage is instead position dependent and occurs at a fixed distance from the axis of symmetry and the conserved sequence of box 3 in the different replicated telomere substrates. This positional dependence for DNA cleavage has not been observed previously for a tyrosine recombinase. PMID- 12753186 TI - IcsB, secreted via the type III secretion system, is chaperoned by IpgA and required at the post-invasion stage of Shigella pathogenicity. AB - Shigella deliver a subset of effector proteins such as IpaA, IpaB and IpaC via the type III secretion system (TTSS) into host cells during the infection of colonic epithelial cells. Many bacterial effectors including some from Shigella require specific chaperones for protection from degradation and targeting to the TTSS. In this study, we have investigated the role of the icsB gene located upstream of the ipaBCDA operon in Shigella infection because the role of IcsB as a virulence factor remains unknown. Here, we found that the IcsB protein is secreted via the TTSS of Shigella in vitro and in vivo. We show that IpgA protein encoded by ipgA, the gene immediately downstream of icsB, serves as the chaperone required for the stabilization and secretion of IcsB. We have shown that IcsB binds to IpgA in bacterial cytosol and the binding site is in the middle of the IcsB protein. Intriguingly, although its significance in Shigella pathogenicity is as yet unclear, the icsB gene can be read-through into the ipgA gene to create a translational fusion protein. Furthermore, the contribution of IcsB to the pathogenicity of Shigella was demonstrated by plaque-forming assay and the Sereny test. The ability of the icsB mutant to form plaques was greatly reduced compared with that of the wild type in MDCK cell monolayers. Furthermore, when guinea pig eyes were infected with a non-polar icsB mutant, the bacteria failed to provoke keratoconjunctivitis. These results suggest that IcsB is secreted via the TTSS, chaperoned by IpgA, and required at the post-invasion stage of Shigella pathogenicity PMID- 12753187 TI - Development and maturation of Escherichia coli K-12 biofilms. AB - The development and maturation of E. coli biofilms in flow-chambers was investigated. We found that the presence of transfer constitutive IncF plasmids induced biofilm development forming structures resembling those reported for Pseudomonas aeruginosa. The development occurred in a step-wise process: (i). attachment of cells to the substratum, (ii). clonal growth and microcolony formation, and (iii). differentiation into expanding structures rising 70-100 microm into the water phase. The first two steps were the same in the plasmid carrying and plasmid-free strains, whereas the third step only occurred in conjugation pilus proficient plasmid-carrying strains. The final shapes of the expanding structures in the mature biofilm seem to be determined by the pilus configuration, as various mutants affected in the processing and activity of the transfer pili displayed differently structured biofilms. We further provide evidence that flagella, type 1 fimbriae, curli and Ag43 are all dispensable for the observed biofilm maturation. In addition, our results indicate that cell-to cell signalling mediated by autoinducer 2 (AI-2) is not required for differentiation of E. coli within a biofilm community. We suggest on the basis of these results that E. coli K-12 biofilm development and maturation is dependent on cell-cell adhesion factors, which may act as inducers of self-assembly processes that result in differently structured biofilms depending on the adhesive properties on the cell surface. PMID- 12753188 TI - Fodrin CaM-binding domain cleavage by Pet from enteroaggregative Escherichia coli leads to actin cytoskeletal disruption. AB - We have previously shown that the plasmid-encoded toxin (Pet) of enteroaggregative Escherichia coli produces cytotoxic and enterotoxic effects. Pet-intoxicated epithelial cells reveal contraction of the cytoskeleton and loss of actin stress fibres. Pet effects require its internalization into epithelial cells. We have also shown that Pet degrades erythroid spectrin. Pet delivery within the intestine suggests that Pet may degrade epithelial fodrin (non erythroid spectrin). Here we demonstrate that Pet has affinity for alpha-fodrin (formally named alphaII spectrin) in vitro and in vivo and cleaves epithelial fodrin, causing its redistribution within the cells. When Pet has produced its cytoskeletal effects, fodrin is found in intracellular aggregates as membrane blebs. Pet cleaves recombinant GST-fodrin, generating two breakdown products of 37 and 72 kDa. Sequencing of the 37 kDa fragment demonstrated that the cleavage site occurred within fodrin's 11th repetitive unit between M1198 and V1199, in the calmodulin binding domain. Site-directed mutagenesis of these amino acids prevented fodrin degradation by Pet. Pet also cleaves epithelial fodrin from cultured Pet-treated cells. A mutant in the Pet serine protease motif was unable to cause fodrin redistribution or to cleave GST-fodrin. This is the first report showing cleavage of alpha-fodrin by a bacterial protease. Cleavage occurs in the middle of the calmodulin binding domain, which leads to cytoskeleton disruption. PMID- 12753189 TI - Calcineurin A of Candida albicans: involvement in antifungal tolerance, cell morphogenesis and virulence. AB - The azole antifungal fluconazole possesses only fungistatic activity in Candida albicans and, therefore, this human pathogen is tolerant to this agent. However, tolerance to fluconazole can be inhibited when C. albicans is exposed to fluconazole combined with the immunosuppressive drug cyclosporin A, which is known to inhibit calcineurin activity in yeast. A mutant lacking both alleles of a gene encoding the calcineurin A subunit (CNA) lost viability in the presence of fluconazole, thus making calcineurin essential for fluconazole tolerance. Consistent with this observation, tolerance to fluconazole was modulated by calcium ions or by the expression of a calcineurin A derivative autoactivated by the removal of its C-terminal inhibitory domain. Interestingly, CNA was also essential for tolerance to other antifungal agents (voriconazole, itraconazole, terbinafine, amorolfine) and to several other metabolic inhibitors (caffeine, brefeldin A, mycophenolic acid, fluphenazine) or cell wall-perturbing agents (SDS, calcofluor white, Congo red), thus indicating that the calcineurin pathway plays an important role in the survival of C. albicans in the presence of external growth inhibitors. Several genes, including PMC1, a vacuolar calcium P type ATPase, were regulated in a calcineurin- and fluconazole-dependent manner. However, PMC1 did not play a direct role in the survival of C. albicans when exposed to fluconazole. In addition to these different properties, calcineurin was found to affect colony morphology in several media known to modulate the C. albicans dimorphic switch. In particular, calcineurin was found to be essential for C. albicans viability in serum-containing media. Finally, calcineurin was found to be necessary for the virulence of C. albicans in a mice model of infection, thus making calcineurin an important element for adequate adaptation to the conditions of the host environment. PMID- 12753190 TI - Evidence that acetyl phosphate functions as a global signal during biofilm development. AB - We used DNA macroarray analysis to identify genes that respond to the status of the intracellular acetyl phosphate (acP) pool. Genes whose expression correlated negatively with the ability to synthesize acP (i.e. negatively regulated genes) function primarily in flagella biosynthesis, a result consistent with observations that we published previously (Pruss and Wolfe, 1994, Mol Microbiol 12: 973-984). In contrast, genes whose expression correlated positively with the ability to synthesize acP (i.e. positively regulated genes) include those for type 1 pilus assembly, colanic acid (capsule) biosynthesis and certain stress effectors. To our knowledge, this constitutes the first report that these genes may respond to the status of the intracellular acP pool. Previously, other researchers have implicated flagella, type 1 pili, capsule and diverse stress effectors in the formation of biofilms. We therefore tested whether cells altered in their ability to metabolize acP could construct normal biofilms, and found that they could not. Cells defective for the production of acP and cells defective for the degradation of acP could both form biofilms, but these biofilms exhibited characteristics substantially different from each other and from biofilms formed by their wild-type parent. We confirmed the role of individual cell surface structures, the expression of which appears to correlate with acP levels, in fim or fli mutants that cannot assemble type 1 pili or flagella respectively. Thus, the information gained by expression profiling of cells with altered acP metabolism indicates that acP may help to co-ordinate the expression of surface structures and cellular processes involved in the initial stages of wild-type biofilm development. PMID- 12753191 TI - PY54, a linear plasmid prophage of Yersinia enterocolitica with covalently closed ends. AB - PY54 is a temperate phage isolated from Yersinia enterocolitica. Lysogenic Yersinia strains harbour the PY54 prophage as a plasmid (pY54). The plasmid has the same size (46 kb) as the PY54 genome isolated from phage particles. By electron microscopy, restriction analysis and DNA sequencing, it was demonstrated that the phage and the plasmid DNAs are linear, circularly permuted molecules. Unusually for phages of Gram-negative bacteria, the phage genome has 3' protruding ends. The linear plasmid pY54 has covalently closed ends forming telomere-like hairpins. The equivalent DNA sequence of the phage genome is a 42 bp perfect palindrome. Downstream from the palindrome, an open reading frame (ORF) was identified that revealed strong DNA homology to the telN gene of Escherichia coli phage N15 encoding a protelomerase. Similar to PY54, the N15 prophage is a linear plasmid with telomeres. The N15 protelomerase has cleaving/joining activity generating the telomeres by processing a 56 bp palindrome (telomere resolution site tel RL). To study the activity of the PY54 protein, the telN-like gene was cloned and expressed in E. coli. A 77 kDa protein was obtained and partially purified. The protein was found to process recombinant plasmids containing the 42 bp palindrome. Telomere resolution of plasmids under in vivo conditions was also investigated in Yersinia infected with PY54. Processing required a plasmid containing the palindrome as well as adjacent DNA sequences from the phage including an additional inverted repeat. Regions on the phage genome important for plasmid maintenance were defined by the construction of linear and circular miniplasmid derivatives of pY54, of which the smallest miniplasmid comprises a 4.5 kb DNA fragment of the plasmid prophage. PMID- 12753192 TI - Suppression of defective ribosome assembly in a rbfA deletion mutant by overexpression of Era, an essential GTPase in Escherichia coli. AB - Era is a small GTP-binding protein and essential for cell growth in Escherichia coli. It consists of two domains: N-terminal GTP-binding and C-terminal RNA binding KH domains. It has been shown to bind to 16S rRNAs and 30S ribosomal subunits in vitro. Here, we report that a precursor of 16S rRNA accumulates in Era-depleted cells. The accumulation of the precursors is also seen in a cold sensitive mutant, E200K, in which the mutation site is located in the C-terminal domain. The major precursor molecule accumulated seems to be 17S rRNA, containing extra sequences at both 5' and 3' ends of 16S rRNA. Moreover, the amounts of both 30S and 50S ribosomal subunits relative to the amount of 70S monosomes increase in Era-depleted and E200K mutant cells. The C-terminal KH domain has a high structural similarity to the RbfA protein, a cold shock protein that also specifically associates with 30S ribosomal subunits. RbfA is essential for cell growth at low temperature, and a precursor of 16S rRNA accumulates in an rbfA deletion strain. The 16S rRNA precursor seems to be identical in size to that accumulated in Era mutant cells. Surprisingly, the cold-sensitive cell growth of the rbfA deletion cells was partially suppressed by overproduction of the wild type Era. The C-terminal domain alone was not able to suppress the cold-sensitive phenotype, whereas Era-dE, which has a 10-residue deletion in a putative effector region of the N-terminal domain, functioned as a more efficient suppressor than the wild-type Era. It was found that Era-dE suppressed defective 16S rRNA maturation, resuming a normal polysome profile to reduce highly accumulated free 30S and 50S subunits in the rbfA deletion cells. These results indicate that Era is involved in 16S rRNA maturation and ribosome assembly. PMID- 12753193 TI - Antagonism of PII signalling by the AmtB protein of Escherichia coli. AB - Escherichia coli AmtB is a member of the MEP/Amt family of ammonia transporters found in archaea, eubacteria, fungi, plants and animals. In prokaryotes, AmtB homologues are co-transcribed with a PII paralogue, GlnK, in response to nitrogen limitation. Here, we show that AmtB antagonizes PII signalling through NRII and that co-expression of GlnK with AmtB overcomes this antagonism. In cells lacking GlnK, expression of AmtB during nitrogen starvation prevented deinduction of Ntr gene expression when a nitrogen source became available. The absence of AmtB in cells lacking GlnK allowed rapid reduction of Ntr gene expression during this transition, indicating that one function of GlnK is to prevent AmtB-mediated antagonism of PII signalling after nitrogen starvation. Other roles of GlnK in controlling Ntr gene expression and maintaining viability during nitrogen starvation were unaffected by AmtB. Expression of AmtB from a constitutive promoter under nitrogen-rich conditions induced full expression of glnALG and elevated expression of glnK in wild-type and glnK cells; thus, the ability of AmtB to raise Ntr gene expression did not require a factor found only in nitrogen starved cells. Experiments with intact cells showed that AmtB acted downstream of a uridylyl transferase uridylyl-removing enzyme (UTase/UR) and upstream of NRII, suggesting that the target was PII. AmtB also slowed the deuridylylation of PII approximately UMP upon ammonia addition, showing that multiple PII interactions were affected by AmtB. Our data are consistent with a hypothesis that AmtB interacts with PII and GlnK, and that co-transcription of glnK and amtB prevents titration of PII when AmtB is highly expressed. PMID- 12753194 TI - Medicinal genetics approach towards identifying the molecular target of a novel inhibitor of fungal cell wall assembly. AB - Glycosylphosphatidylinositol (GPI)-anchored cell wall mannoproteins are required for the adhesion of pathogenic fungi, such as Candida albicans, to human epithelium. Small molecular inhibitors of the cell surface presentation of GPI anchored mannoproteins would be promising candidate drugs to block the establishment of fungal infections. Here, we describe a medicinal genetics approach to identifying the gene encoding a novel target protein that is required for the localization of GPI-anchored cell wall mannoproteins. By means of a yeast cell-based screening procedure, we discovered a compound, 1-[4 butylbenzyl]isoquinoline (BIQ), that inhibits cell wall localization of GPI anchored mannoproteins in Saccharomyces cerevisiae. Treatment of C. albicans cells with this compound resulted in reduced adherence to a rat intestine epithelial cell monolayer. A previously uncharacterized gene YJL091c, named GWT1, was cloned as a dosage-dependent suppressor of the BIQ-induced phenotypes. GWT1 knock-out cells showed similar phenotypes to BIQ-treated wild-type cells in terms of cell wall structure and transcriptional profiles. Two different mutants resistant to BIQ each contained a single missense mutation in the coding region of the GWT1 gene. These results all suggest that the GWT1 gene product is the primary target of the compound. PMID- 12753195 TI - Substrate specificity of type III flagellar protein export in Salmonella is controlled by subdomain interactions in FlhB. AB - FlhB, an integral membrane protein, gates the type III flagellar export pathway of Salmonella. It permits export of rod/hook-type proteins before hook completion, whereupon it switches specificity to recognize filament-type proteins. The cytoplasmic C-terminal domain of FlhB (FlhBC) is cleaved between Asn-269 and Pro-270, defining two subdomains: FlhBCN and FlhBCC. Here, we show that subdomain interactions and cleavage within FlhB are central to substrate specificity switching. We found that deletions between residues 216 and 240 of FlhBCN permitted FlhB cleavage but abolished function, whereas a deletion spanning Asn-269 and Pro-270 abolished both. The mutation N269A prevented cleavage at the FlhBCN-FlhBCC boundary. Cells producing FlhB(N269A) exported the same amounts of hook-capping protein as cells producing wild-type FlhB. However, they exported no flagellin, even when the fliC gene was being expressed from a foreign promoter to circumvent regulation of expression by FlgM, which is itself a filament-type substrate. Electron microscopy revealed that these cells assembled polyhook structures lacking filaments. Thus, FlhB(N269A) is locked in a conformation specific for rod/hook-type substrates. With FlhB(P270A), cleavage was reduced but not abolished, and cells producing this protein were weakly motile, exported reduced amounts of flagellin and assembled polyhook filaments. PMID- 12753196 TI - A LuxR-type regulator from Agrobacterium tumefaciens elevates Ti plasmid copy number by activating transcription of plasmid replication genes. AB - TraR, a LuxR-type quorum-sensing transcription factor in Agrobacterium tumefaciens, activates genes required for conjugal transfer of the Ti plasmid and also enhances the copy number of a nopaline-type Ti plasmid. Here, we show that TraR increases the copy number of an octopine-type Ti plasmid up to eightfold and that TraR activates transcription of the repABC operon up to 25-fold. The ability of TraR to increase copy number was strictly dependent on several TraR-activated promoters of this operon, indicating that TraR affects copy number solely at the level of transcription. Promoter resections and mRNA transcript analysis revealed the presence of three TraR-dependent promoters. Two TraR-dependent transcription start sites are located 45.5 and 65.5 nucleotides downstream of a site called tra box II, whereas the third start site lies 42.5 nucleotides downstream of a site called tra box III. Purified TraR bound to both tra boxes with comparable affinities, causing moderate DNA bending. TraR bound and bent these two sites independently rather than synergistically. Alteration of tra box III to match the consensus sequence dramatically increased TraR-dependent expression of repABC and plasmid copy number. TraR-dependent elevation of Ti plasmid copy number caused a three- to fourfold increase in plant tumorigenesis. PMID- 12753197 TI - SarA and not sigmaB is essential for biofilm development by Staphylococcus aureus. AB - Staphylococcus aureus biofilm formation is associated with the production of the polysaccharide intercellular adhesin (PIA/PNAG), the product of the ica operon. The staphylococcal accessory regulator, SarA, is a central regulatory element that controls the production of S. aureus virulence factors. By screening a library of Tn917 insertions in a clinical S. aureus strain, we identified SarA as being essential for biofilm development. Non-polar mutations of sarA in four genetically unrelated S. aureus strains decreased PIA/PNAG production and completely impaired biofilm development, both in steady state and flow conditions via an agr-independent mechanism. Accordingly, real-time PCR showed that the mutation in the sarA gene resulted in downregulation of the ica operon transcription. We also demonstrated that complete deletion of sigmaB did not affect PIA/PNAG production and biofilm formation, although it slightly decreased ica operon transcription. Furthermore, the sarA-sigmaB double mutant showed a significant decrease of ica expression but an increase of PIA/PNAG production and biofilm formation compared to the sarA single mutant. We propose that SarA activates S. aureus development of biofilm by both enhancing the ica operon transcription and suppressing the transcription of either a protein involved in the turnover of PIA/PNAG or a repressor of its synthesis, whose expression would be sigmaB-dependent. PMID- 12753198 TI - Construction and usage of a onefold-coverage shotgun DNA microarray to characterize the metabolism of the archaeon Haloferax volcanii. AB - Haloferax volcanii is a moderately halophilic archaeon that can grow aerobically and anaerobically with a variety of substrates. We undertook a novel approach for the characterization of metabolic adaptations, i.e. transcriptome analysis with a onefold-coverage shotgun DNA microarray. A genomic library was constructed and converted into a polymerase chain reaction (PCR) product library, which was used to print two DNA microarrays, a 960-spot test array used for optimization of microarray analysis and a 2880-spot onefold-coverage array. H. volcanii cultures were shifted from casamino acid-based metabolism to glucose-based metabolism, and the transcriptome changes were analysed with the onefold-coverage array at five time points covering the transition phase and the onset of exponential growth with the new carbon source. About 10% of all genes were found to be more than 2.5 fold regulated at at least one time point. The genes fall into five clusters of kinetically co-regulated genes. For members of all five clusters, the results were verified by Northern blot analyses. The identity of the regulated genes was determined by sequencing. Many co-regulated genes encode proteins of common functions. Expected as well as a variety of unexpected findings allowed predictions about the central metabolism, the transport capacity and the cellular composition of H. volcanii growing on casamino acids and on glucose. The microarray analyses are in accordance with the growth rates and ribosome contents of H. volcanii growing on the two carbon sources. Analysis of the results revealed that onefold-coverage shotgun DNA microarrays are well suited to characterize the regulation of metabolic pathways as well as protein complexes in response to changes in environmental conditions. PMID- 12753199 TI - Mechanisms of homology-facilitated illegitimate recombination for foreign DNA acquisition in transformable Pseudomonas stutzeri. AB - Intra- and interspecific natural transformation has been observed in many prokaryotic species and is considered a fundamental mechanism for the generation of genetic variation. Recently, it has been described in detail how, in transformable Acinetobacter BD413 and Streptococcus pneumoniae, long stretches of nucleotides lacking homology were integrated into recipient genomes when they were linked on one side to a small piece of DNA with homology to resident DNA serving as a recA-dependent recombination anchor. Now, such homology-facilitated illegitimate recombination (HFIR) has also been detected in transformable Pseudomonas stutzeri. However, analysis of the recombinants revealed qualitative and quantitative differences in their generation compared with that in Acinetobacter BD413. In P. stutzeri, foreign DNA with an anchor sequence was integrated 105- to 106-fold less frequently than fully homologous DNA, but still at least 200-fold more frequently than without the anchor. The anchor sequence could be as small as 311 bp. Remarkably, in 98% of the events, the 3' end was integrated within the homologous anchor, whereas the 5' end underwent illegitimate fusion. Moreover, about one-third of the illegitimate fusion sites shared no or only a single identical basepair in foreign and resident DNA. The other fusions occurred within microhomologies of up to 6 bp with a higher GC content on average than the interacting nucleotide sequences. Foreign DNA of 69 1903 bp was integrated, and resident DNA of 22-2345 bp was lost. In a recA mutant, HFIR was not detectable. The findings suggest that genomic acquisition of foreign DNA by HFIR during transformation occurs widely in prokaryotes, but that details of the required recombination and strand fusion mechanisms may differ between organisms from different genera. PMID- 12753200 TI - TFIIB and subunits of the SAGA complex are involved in transcriptional activation of phospholipid biosynthetic genes by the regulatory protein Ino2 in the yeast Saccharomyces cerevisiae. AB - In the yeast Saccharomyces cerevisiae, genes involved in phospholipid biosynthesis are activated by ICRE (inositol/choline-responsive element) up stream motifs and the corresponding heterodimeric binding factor, Ino2 + Ino4. Both Ino2 and Ino4 contain basic helix-loop-helix (bHLH) domains required for ICRE binding, whereas transcriptional activation is mediated exclusively by Ino2. In this work, we describe a molecular analysis of functional minimal domains responsible for specific DNA recognition and transcriptional activation (TAD1 and TAD2). We also define the importance of individual amino acids within the more important activation domain TAD1. Random mutagenesis at five amino acid positions showed the importance of acidic as well as hydrophobic residues within this minimal TAD. We also investigated the contribution of known general transcription factors and co-activators for Ino2-dependent gene activation. Although an ada5 single mutant and a gal11 paf1 double mutant were severely affected, a partial reduction in activation was found for gcn5 and srb2. Ino2 interacts physically with the basal transcription factor Sua7 (TFIIB of yeast). Interestingly, interaction is mediated by the HLH dimerization domain of Ino2 and by two non overlapping domains within Sua7. Thus, Sua7 may compete with Ino4 for binding to the Ino2 activator, creating the possibility of positive and negative influence of Sua7 on ICRE-dependent gene expression. PMID- 12753201 TI - Dual regulation by phospho-OmpR of ssrA/B gene expression in Salmonella pathogenicity island 2. AB - Expression of genes located on Salmonella pathogenicity island 2 (SPI-2) is required for systemic infection in mice. This region encodes a type III secretion system, secreted effectors and the two-component regulatory system SsrA/B (also referred to as SpiR), as well as additional uncharacterized genes. In the present work, we demonstrate that phospho-OmpR (OmpR-P) functions as an activator at the spiC-ssrA/B locus. There are two promoters at spiR; one is upstream of ssrA and the other upstream of ssrB. Our results indicate that, in contrast to many two component regulatory systems, regulation of the sensor kinase SsrA appears to be uncoupled and distinct from regulation of the response regulator SsrB. OmpR regulation of ssrA/B is one of only a few examples known in which a two-component response regulator directly regulates the expression of another two-component regulatory system. PMID- 12753202 TI - A 'fair go' for coral hybridization. AB - Hybridisation between coral species clearly occurs in vitro, but the evolutionary significance of this cross-fertility is still the subject of much debate. Compelling genetic and reproductive evidence support introgressive hybridization amongst Indo-Pacific members of the scleractinian genus Acropora. Although population genetic analyses indicate that interspecific hybridization events are relatively rare, they are likely be important on evolutionary time scales, creating the capacity for adaptive evolution by increasing genomic diversity and heterozygosity. However, in a recent paper based exclusively on the three endemic Caribbean Acropora species, Vollmer and Palumbi (2002) dispute the occurrence of reticulation in corals. Here we use data from both the Vollmer and Palumbi study and our earlier paper on the same species (van Oppen et al., 2000) to show that reticulation has occurred amongst the Caribbean Acropora species. Furthermore, conclusions based on the limited Caribbean Acropora fauna cannot simply be extrapolated to Indo-Pacific corals, and it is inappropriate to view some coral species as 'immortal mules'. PMID- 12753203 TI - Clonal diversity and genetic differentiation in Ilex leucoclada M. patches in an old-growth beech forest. AB - We investigated clonal diversity within patches of Ilex leucoclada and genetic variation within and among patches using random amplified polymorphic DNA (RAPD) markers in a 1-ha plot within an old-growth beech forest. We found 38 patches that exhibited a clumped distribution in the middle of the plot. We identified a total of 166 RAPD phenotypes among the 215 stems sampled from 27 patches that were completely within the plot. The population showed high clonal diversity within patches (mean number of genets relative to number of stems = 0.79; mean Simpson's D = 0.89). Variation in RAPD phenotypes among patches was highly significant (PhiST in the molecular variance analysis = 0.316, P < 0.001), indicating genetic differentiation among patches. Pairwise genetic distances, PhiST, among patches did not correlate with geographical distances among patches. The cluster analysis based on the genetic distances showed few clear clusters of patches, indicating no spatial genetic structure among patches. High levels of clonal diversity both within patches and within the population may be explained by multiple founders, seedling recruitment during patch-formation, and somatic mutation. The significant genetic differentiation among patches may be caused by separate founding events and/or kin structuring within patches. PMID- 12753204 TI - Combining genetic markers and stable isotopes to reveal population connectivity and migration patterns in a neotropical migrant, Wilson's warbler (Wilsonia pusilla). AB - We used results from the analysis of microsatellite DNA variation and hydrogen stable-isotope ratios to characterize the population structure of a neotropical migrant passerine, the Wilson's warbler (Wilsonia pusilla). The resulting information was then used to infer migration patterns and population connectivity between breeding grounds in North America and overwintering areas in Mexico and Central America. The microsatellite data revealed genetic structure across the North American continent; populations in the west were found to significantly differ from the east. Minimal genetic structure was observed among western sites. The lack of isolation by distance and low variance in FST values suggests that gene flow could play an ongoing role in limiting genetic differentiation among sites in the western part of the distribution. However, additional information including estimates of effective population size and the proximity of the population to equilibrium is required before the role of gene flow can be assessed fully. Analysis of isotope data showed a negative relationship between latitude and hydrogen isotope ratios in breeding ground individuals. There was a positive relationship between wintering ground latitude and hydrogen isotope ratios for individuals that were genetically western in origin. This is consistent with a leapfrog pattern of migration, in which genetically western birds from the northernmost breeding areas overwinter at the most southerly locations in Central America. Additionally, isotopic ratios of western birds suggest that coastal breeders overwinter in western Mexico, while western birds from further inland and at high elevations overwinter in eastern Mexico. Using information from both genetic an isotopic approaches will probably be useful for identifying patterns of migration and population connectivity between breeding and overwintering areas, both important issues for conservation efforts, and may also contribute to investigation of the evolution of migration. PMID- 12753206 TI - Genetic variation in the endangered wild apple (Malus sylvestris (L.) Mill.) in Belgium as revealed by amplified fragment length polymorphism and microsatellite markers. AB - The genetic variation within and between wild apple samples (Malus sylvestris) and cultivated apple trees was investigated with amplified fragment length polymorphisms (AFLP) and microsatellite markers to develop a conservation genetics programme for the endangered wild apple in Belgium. In total, 76 putative wild apples (originating from Belgium and Germany), six presumed hybrids and 39 cultivars were typed at 12 simple sequence repeats (SSR) and 139 amplified fragment length polymorphism (AFLP) loci. Principal co-ordinate analysis and a model-based clustering method classified the apples into three major gene pools: wild Malus sylvestris genotypes, edible cultivars and ornamental cultivars. All presumed hybrids and two individuals (one Belgian, one German) sampled as M. sylvestris were assigned completely to the edible cultivar gene pool, revealing that cultivated genotypes are present in the wild. However, gene flow between wild and cultivated gene pools is shown to be almost absent, with only three genotypes that showed evidence of admixture between the wild and edible cultivar gene pools. Wild apples sampled in Belgium and Germany constitute gene pools that are clearly differentiated from cultivars and although some geographical pattern of genetic differentiation among wild apple populations exists, most variation is concentrated within samples. Concordant conclusions were obtained from AFLP and SSR markers, which showed highly significant correlations in both among-genotypes and among-samples genetic distances. PMID- 12753205 TI - Ancestral polymorphisms in genetic markers obscure detection of evolutionarily distinct populations in the endangered Florida grasshopper sparrow (Ammodramus savannarum floridanus). AB - Genetic analyses of bird subspecies designated as conservation units can address whether they represent units with independent evolutionary histories and provide insights into the evolutionary processes that determine the degree to which they are genetically distinct. Here we use mitochondrial DNA control region sequence and six microsatellite DNA loci to examine phylogeographical structure and genetic differentiation among five North American grasshopper sparrow (Ammodramus savannarum) populations representing three subspecies, including a population of the endangered Florida subspecies (A. s. floridanus). This federally listed taxon is of particular interest because it differs phenotypically from other subspecies in plumage and behaviour and has also undergone a drastic decline in population size over the past century. Despite this designation, we observed no phylogeographical structure among populations in either marker: mtDNA haplotypes and microsatellite genotypes from floridanus samples did not form clades that were phylogenetically distinct from variants found in other subspecies. However, there was low but significant differentiation between Florida and all other populations combined in both mtDNA (FST = 0.069) and in one measure of microsatellite differentiation (theta = 0.016), while the non-Florida populations were not different from each other. Based on analyses of mtDNA variation using a coalescent-based model, the effective sizes of these populations are large (approximately 80,000 females) and they have only recently diverged from each other (< 26,000 ybp). These populations are probably far from genetic equilibrium and therefore the lack of phylogenetic distinctiveness of the floridanus subspecies and minimal genetic differentiation is due most probably to retained ancestral polymorphism. Finally, levels of variation in Florida were similar to other populations supporting the idea that the drastic reduction in population size which has occurred within the last 100 years has not yet had an impact on levels of variation in floridanus. We argue that despite the lack of phylogenetic distinctiveness of floridanus genotypes the observed genetic differentiation and previously documented phenotypic differences justify continued designation of this subspecies as a protected population segment. PMID- 12753207 TI - Nuclear gene sequences and DNA variation of Cryptomeria japonica samples from the postglacial period. AB - Genomic DNA was extracted from heartwood blocks of six Cryptomeria japonica individuals that had been buried (in an area now covered by rice fields) for about 3600 years. Attempts were made to determine the sequences of five nuclear genes following polymerase chain reaction amplification, using previously obtained C. japonica expressed sequence tag (EST) information. We detected 15 nucleotide substitutions and four insertion/deletions (indels) in a partial GapC gene sequence among 13 individuals of the buried and an extant population, which allowed us to estimate the extent of DNA variation within the buried populations, and the level of genetic differentiation between the buried population and the extant population growing in a neighbouring area. For the entire haplotypes of the GapC region, pi and theta nucleotide diversity estimates were 0.0063 and 0.0010, respectively, when both populations were included, while corresponding figures for the buried population alone were 0.0009 and 0.0017. Estimates of DNA divergence statistics (dXY = 0.0062, dA = 0.0005, FST = 0.0832 and KST = 0.0935) suggest that differentiation between the two populations was not great. However, permutation tests gave FST and KST values rejecting the null hypothesis (that populations were not differentiated) at the 5% and 1% probability levels, respectively. The significant genetic differentiation between the two populations was mainly caused by differences in haplotype diversity. The significant level of haplotype diversity in the extant population compared to the buried population might be the result of gene flow from neighbouring artificial forests. Alternatively, it is possible that we failed to detect all the DNA variation in the buried population because of clonal growth in the buried population. PMID- 12753208 TI - Two centuries of the Scandinavian wolf population: patterns of genetic variability and migration during an era of dramatic decline. AB - The grey wolf (Canis lupus) was numerous on the Scandinavian peninsula in the early 19th century. However, as a result of intense persecution, the population declined dramatically and was virtually extinct from the peninsula by the 1960s. We examined historical patterns of genetic variability throughout the period of decline, from 1829 to 1979. Contemporary Finnish wolves, considered to be representative of a large eastern wolf population, were used for comparison. Mitochondrial DNA (mtDNA) variability among historical Scandinavian wolves was significantly lower than in Finland while Y chromosome variability was comparable between the two populations. This may suggest that long-distance migration from the east has been male-biased. Importantly though, as the historical population was significantly differentiated from contemporary Finnish wolves, the overall immigration rate to the Scandinavian peninsula appears to have been low. Levels of variability at autosomal microsatellite loci were high by the early 1800s but declined considerably towards the mid-20th century. At this time, approximately 40% of the allelic diversity and 30% of the heterozygosity had been lost. After 1940, however, there is evidence of several immigration events, coinciding with episodes of marked population increase in Russian Karelia and subsequent westwards migration. PMID- 12753209 TI - Hierarchical spatial structure of genetically variable nucleopolyhedroviruses infecting cyclic populations of western tent caterpillars. AB - The cyclic population dynamics of western tent caterpillars, Malacosoma californicum pluviale, are associated with epizootics of a nucleopolyhedrovirus, McplNPV. Given the dynamic fluctuations in host abundance and levels of viral infection, host resistance and virus virulence might be expected to change during different phases of the cycle. As a first step in determining if McplNPV virulence and population structure change with host density, we used restriction fragment length polymorphism (RFLP) analysis to examine the genetic diversity of McplNPV infecting western tent caterpillar populations at different spatial scales. Thirteen dominant genetic variants were identified in 39 virus isolates (individual larvae) collected from field populations during one year of low host density, and another distinct variant was discovered among nine additional isolates in two subsequent years of declining host density. The distribution of these genetic variants was not random and indicated that the McplNPV population was structured at several spatial levels. A high proportion of the variation could be explained by family grouping, which suggested that isolates collected within a family were more likely to be the same than isolates compared among populations. Additionally, virus variants from within populations (sites) were more likely to be the same than isolates collected from tent caterpillar populations on different islands. This may indicate that there is limited mixing of virus among tent caterpillar families and populations when host population density is low. Thus there is potential for the virus to become locally adapted to western tent caterpillar populations in different sites. However, no dominant genotype was observed at any site. Whether and how selection acts on the genetically diverse nucleopolyhedrovirus populations as host density changes will be investigated over the next cycle of tent caterpillar populations. PMID- 12753211 TI - A test for deviation from island-model population structure. AB - The neutral island model forms the basis for several estimation models that relate patterns of genetic structure to microevolutionary processes. Estimates of gene flow are often based on this model and may be biased when the model's assumptions are violated. An appropriate test for violations is to compare FST scores for individual loci to a null distribution based on the average FST taken over multiple loci. A parametric bootstrap method is described here based on Wright's beta-distribution to generate null distributions of FST for each locus. These null distributions account for error introduced by sampling populations, individuals and loci, and also biological sources of error, including variable alleles/locus and inbreeding. Confidence limits can be obtained directly from these distributions. Significant deviations from the island model may be the result of selection, deviations from the island model's migration pattern, nonequilibrium conditions, or other deviations from island-model assumptions. Only strong biases are likely to be detected because of the inherently large sampling variation of FST. Nevertheless, a coefficient, Nb, describing bias in the spread of the beta-distribution in units comparable to the gene flow parameter, Nm, can be obtained for each locus. In samples from populations of the butterfly Coenonympha tullia, the loci Idh-1, Mdh-1, Pgi and Pgm showed significantly lower FST than expected. PMID- 12753210 TI - Diverging patterns of mitochondrial and nuclear DNA diversity in subarctic black spruce: imprint of a founder effect associated with postglacial colonization. AB - High-latitude ecotonal populations at the species margins may exhibit altered patterns of genetic diversity, resulting from more or less recent founder events and from bottleneck effects in response to climate oscillations. Patterns of genetic diversity were investigated in nine populations of the conifer black spruce (Picea mariana [Mill.] BSP.) in northwestern Quebec, Canada, using seed dispersed mitochondrial (mt) DNA and nuclear (nc) DNA. mtDNA diversity (mitotypes) was assessed at three loci, and ncDNA diversity was estimated for nine expressed sequence tag polymorphism (ESTP) loci. Sampling included populations from the boreal forest and the southern and northern subzones of the subarctic forest-tundra, a fire-born ecotone. For ncDNA, populations from all three vegetation zones were highly diverse with little population differentiation (thetaN = 0.014); even the northernmost populations showed no loss of rare alleles. Patterns of mitotype diversity were strikingly different: within population diversity and population differentiation were high for boreal forest populations [expected heterozygosity per locus (HE) = 0.58 and thetaM = 0.529], but all subarctic populations were fixed for a single mitotype (HE = 0). This lack of variation suggests a founder event caused by long-distance seed establishment during postglacial colonization, consistent with palaeoecological data. The estimated movement of seeds alone (effective number of migrants per generation, NmM < 2) was much restricted compared to that estimated from nuclear variants, which including pollen movement (NmN > 17). This could account for the conservation of a founder imprint in the mtDNA of subarctic black spruce. After reduction, presumably in the early Holocene, the diversity in ncDNA would have been replenished rapidly by pollen-mediated gene flow, and maintained subsequently through vegetative layering during the current cooler period covering the last 3000 years. PMID- 12753212 TI - Distribution and diversity of rhizobia nodulating agroforestry legumes in soils from three continents in the tropics. AB - The natural rhizobial populations of Calliandra calothyrsus, Gliricidia sepium, Leucaena leucocephala and Sesbania sesban were assessed in soils from nine sites across tropical areas of three continents. The rhizobial population size varied from undetectable numbers to 1.8 x 104 cells/g of soil depending on the trap host and the soil. Calliandra calothyrsus was the most promiscuous legume, nodulating in eight soils, while S. sesban nodulated in only one of the soils. Polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) analyses of the 16S rRNA gene and the internally transcribed spacer (ITS) region between the 16S and 23S rRNA genes were used to assess the diversity and relative abundance of rhizobia trapped from seven of the soils by C. calothyrsus, G. sepium and L. leucocephala. Representatives of the 16S rRNA RFLP groups were also subjected to sequence analysis of the first 950 base pairs of the 16S rRNA gene. Eighty ITS groups were obtained, with none of the ITS types being sampled in more than one soil. RFLP analysis of the 16S rRNA yielded 23 'species' groups distributed among the Rhizobium, Mesorhizobium, Sinorhizobium and Agrobacterium branches of the rhizobial phylogenetic tree. The phylogeny of the isolates was independent of the site or host of isolation, with different rhizobial groups associated with each host across the soils from widely separated geographical regions. Although rhizobial populations in soils sampled from the centre of diversity of the host legumes were the most genetically diverse, soil acidity was highly correlated with the diversity of ITS types. Our results support the hypothesis that the success of these tree legumes in soils throughout the tropics is the result of their relative promiscuity (permissiveness) allowing nodulation with diverse indigenous rhizobial types. PMID- 12753213 TI - Differential cycles of range contraction and expansion in European high mountain plants during the Late Quaternary: insights from Pritzelago alpina (L.) O. Kuntze (Brassicaceae). AB - Nuclear DNA sequence variation of the internal transcribed spacer (ITS) and amplified fragment length polymorphisms (AFLPs) were used to illuminate the evolutionary history of Pritzelago alpina, a herbaceous perennial of (sub)alpine to nival habitats of the European high mountains. Maximum likelihood analysis of ITS sequences of P. alpina, Hornungia petraea and Hymenolobus procumbens (the 'Pritzelago alliance') resolved P. alpina and H. petraea as sister taxa. ITS divergence estimates support an origin for P. alpina in the Late Tertiary, while intraspecific diversification started in the Late Quaternary (0.4-0.9 million years ago). AFLP analysis of 76 individuals of P. alpina, representing 24 localities across its entire west-east distribution, identified four mountain lineages in Cantabria, the Pyrenees, (south-) western Alps, and northeastern Alps/Tatras/Carpathians. In an analysis of molecular variance (amova), 14.3% of the total variation derived from this separation. However, relationships among these lineages remained unresolved in neighbour-joining and principal co ordinates analyses, suggesting a population history of near simultaneous vicariance. Comparison with our previous ITS/AFLP study of Anthyllis montana (Fabaceae) indicates that the two co-distributed but altitudinally differentiated plant species exhibit temporally concordant but spatially discordant patterns of genetic variation. Moreover, levels of AFLP divergence were significantly lower in P. alpina than in the submediterranean, lower-elevation A. montana. Together, these data are consistent with a 'displacement refugia model', which predicts that European mountain plant species associated with lower- and upper-elevation habitats had a different cycle of range contraction into (long-term) glacial and (short-term) interglacial refugia, respectively. PMID- 12753214 TI - Phylogeography of the common vole (Microtus arvalis) with particular emphasis on the colonization of the Orkney archipelago. AB - To investigate the human introduction of the common vole Microtus arvalis onto the Orkney islands, the complete cytochrome b gene was sequenced in 41 specimens from both Orkney (four localities) and elsewhere in their range (26 localities). Orkney voles belonged to the same phylogenetic lineage, 'Western', as individuals from France and Spain indicating southwestern Europe as the most likely source area for the islands. This result is of interest with respect to the movement and trading links of the Neolithic people who likely transported the voles. As well as the Western lineage, our phylogenetic trees revealed three other purely European lineages: the 'Italian' (single specimen from N. Italy), the 'Central' (Germany, Netherlands, Denmark) and the 'Eastern' (Hungary, Slovakia, Poland, Ukraine, Finland, European Russia). Individuals from European Russia, W. Siberia, Georgia, Ukraine and Armenia formed a fifth distinct lineage coinciding with the distribution of the 'obscurus' chromosomal form of M. arvalis. These phylogeographical data suggest that M. arvalis occupied multiple refugia during the last glaciation. PMID- 12753215 TI - Holarctic phylogeography of the root vole (Microtus oeconomus): implications for late Quaternary biogeography of high latitudes. AB - A species-wide phylogeographical study of the root vole (Microtus oeconomus) was performed using the whole 1140 base pair mitochondrial (mt) cytochrome b gene. We examined 83 specimens from 52 localities resulting in 65 unique haplotypes. Our results demonstrate that the root vole is divided into four main mtDNA phylogenetic lineages that seem to have largely allopatric distributions. Net divergence estimates (2.0-3.5%) between phylogroups, as well as relatively high nucleotide diversity estimates within phylogroups, indicate that the distinct phylogeographical structure was initiated by historical events that predated the latest glaciation. European root voles are divided into a Northern and a Central mtDNA phylogroup. The mtDNA data in concert with fossil records imply that root voles remained north of the classical refugial areas in southern Europe during the last glacial period. The currently fragmented populations in central Europe belong to a single mtDNA phylogroup. The Central Asian and the North European lineages are separated by the Ural Mountains, a phylogeographical split also found in collared lemmings (Dicrostonyx) and the common vole (M. arvalis). The Beringian lineage occurs from eastern Russia through Alaska to northwestern Canada. This distribution is congruent with the traditional boundaries of the Beringian refugium and with phylogeographical work on other organisms. In conclusion, similarities between the phylogeographical patterns in the root vole and other rodents, such as Arctic and subarctic lemmings, as well as more temperate vole species, indicate that late Quaternary geological and climatic events played a strong role in structuring northern biotic communities. PMID- 12753216 TI - Testing Pleistocene refugia theory: phylogeographical analysis of Desmognathus wrighti, a high-elevation salamander in the southern Appalachians. AB - During the colder climates of the Pleistocene, the ranges of high-elevation species in unglaciated areas may have expanded, leading to increased gene flow among previously isolated populations. The phylogeography of the pygmy salamander, Desmognathus wrighti, an endemic species restricted to the highest mountain peaks of the southern Appalachians, was examined to test the hypothesis that the range of D. wrighti expanded along with other codistributed taxa during the Pleistocene. Analyses of genetic variation at 14 allozymic loci and of the 12S rRNA gene in the mtDNA genome was conducted on individuals sampled from 14 population isolates throughout the range of D. wrighti. In contrast to the genetic patterns of many other high-elevation animals and plants, genetic distances derived from both molecular markers showed significant isolation by distance and genetic structuring of populations, suggesting long-term isolation of populations. Phylogeographical analyses revealed four genetically distinct population clusters that probably remained fragmented during the Pleistocene, although there was also evidence supporting recent gene flow among some population groups. Support for isolation by distance is rare among high-elevation species in unglaciated areas of North and Middle America, although not uncommon among Plethodontid Salamanders, and this pattern suggests that populations of D. wrighti did not expand entirely into suitable habitat during the Pleistocene. We propose that intrinsic barriers to dispersal, such as species interactions with other southern Appalachian plethodontid salamanders, persisted during the Pleistocene to maintain the fragmented distribution of D. wrighti and allow for significant genetic divergence of populations by restricting gene flow. PMID- 12753217 TI - Phylogeography of Ponto-Caspian crustaceans: a benthic-planktonic comparison. AB - The Black, Azov, Caspian and Aral Seas, remnants of the intracontinental Paratethys basin, are home to a spectacular diversity of crustaceans. This study examines the past history of the Ponto-Caspian fauna through comparative phylogeographical studies on both benthic and planktonic taxa, based on an examination of nucleotide diversity in the mitochondrial, cytochrome c oxidase subunit 1 (COI) gene. The COI data reveal a striking example of phylogeographical concordance. All species analysed, three amphipods and three cladocerans, are characterized by two monophyletic clades corresponding to the Black and Caspian regions. However, this phylogeographical partition is, on average, four times deeper for the benthic amphipods than for the planktonic cladocerans. Based on standard molecular clocks, the Black and Caspian lineages of benthic crustaceans diverged at varied intervals from 1 to 8 million years ago. By contrast, planktonic lineages are more recent with their divergence occurring in the last million years. Levels of intraspecific polymorphisms are variable and generally lower in planktonic than benthic taxa. The mechanisms responsible for the high diversity of crustaceans in the Ponto-Caspian region are discussed on the basis of these results. PMID- 12753218 TI - Molecular phylogeography of the Amazonian Chagas disease vectors Rhodnius prolixus and R. robustus. AB - The phylogeographical structure of the closely related species Rhodnius prolixus and R. robustus is presented based on a 663-base pair (bp) fragment of the mitochondrial cytochrome b gene. Twenty haplotypes were recovered from 84 samples examined, representing 26 populations from seven Latin American countries. The resulting phylogenetic tree is composed of five major reciprocally monophyletic clades, one representing R. prolixus and four representing R. robustus. While R. prolixus is a very homogeneous assemblage, R. robustus has deeper clades and is paraphyletic, with the clade comprising R. robustus from Venezuela (Orinoco region) more closely related to the R. prolixus clade than to the other R. robustus populations from the Amazon region. The R. robustus paraphyly was supported further by the analysis of a nuclear gene (D2 region of the 28S RNA) for a subset of specimens. The data support the view that R. robustus represents a species complex. Levels of sequence divergence between clades within each region are compatible with a Pleistocene origin. Nucleotide diversity (pi) for all R. prolixus populations was extremely low (0.0008), suggesting that this species went through a recent bottleneck, and was subsequently dispersed by man. PMID- 12753219 TI - Analysis of molecular data of Arabidopsis thaliana (L.) Heynh. (Brassicaceae) with Geographical Information Systems (GIS). AB - A Geographical Information System (GIS) is used to analyse allelic information of 13 sequenced loci of natural populations of Arabidopsis thaliana and to identify geographical structures. GIS provides tools for visualization and analysis of geographical population structures using molecular data. The geographical distribution of the number of variable positions in the alignments, the distribution of recombinant sequence blocks, and the distribution of a newly defined measure, the differentiation index, are studied. The differentiation index is introduced to measure the sequence divergence among individual plants sampled from various geographical localities. The numbers of variable positions and the differentiation index are also used for a metadata analysis covering about 26 kb of the genome. This analysis reveals, for the first time, differences in DNA sequence structures of geographically different populations of A. thaliana. The broadly defined west Mediterranean region consists of accessions with the highest numbers of polymorphic positions followed by the west European region. The GIS technology Kriging is used to define Arabidopsis specific diversity zones in Europe. The highest genetic variability is observed along the Atlantic coast from the western Iberian Peninsula to southern Great Britain, while lowest variability is found in central Europe. PMID- 12753220 TI - Patterns of molecular evolution and diversification in a biodiversity hotspot: the California Floristic Province. AB - The California Floristic Province harbours more endemic plant and animal taxa and more identifiable subspecies than any other area of comparable size in North America. We present evidence that physical historical processes have resulted in congruent patterns of genetic diversity over the past 2-10 million years. Using a molecular clock approach we show that diversification and establishment of spatial genetic structure across six taxonomic groups coincide with the putative age of California's mountain ranges and aridification in the region. Our results demonstrate the importance of geographical barriers and climatological events to species diversification and the overall geographical structure of biodiversity. These results should facilitate conservation efforts in this biodiversity hotspot for taxa whose population genetic structure is still unknown and may suggest the potential utility of this approach in regional conservation planning efforts. PMID- 12753221 TI - Co-occurrence in nature of different clones of the social amoeba, Dictyostelium discoideum. AB - The social amoeba, Dictyostelium discoideum, produces a multicellular fruiting body and has become a model system for cell-cell interactions such as signalling, adhesion and development. However, unlike most multicellular organisms, it forms by aggregation of cells and, in the laboratory, forms genetic chimeras where there may be competition among clones. Here we show that chimera formation is also likely in nature, because different clones commonly co-occur on a very small scale. This suggests that D. discoideum will likely have evolved strategies for competing in chimeras, and that the function of some developmental genes will be competitive. Natural chimerism also makes D. discoideum a good model organism for the investigation of issues relating to coexistence and conflict between cells. PMID- 12753222 TI - Prospects for inferring pairwise relationships with single nucleotide polymorphisms. AB - An extraordinarily large number of single nucleotide polymorphisms (SNPs) are now available in humans as well as in other model organisms. Technological advancements may soon make it feasible to assay hundreds of SNPs in virtually any organism of interest. One potential application of SNPs is the determination of pairwise genetic relationships in populations without known pedigrees. Although microsatellites are currently the marker of choice for this purpose, the number of independently segregating microsatellite markers that can be feasibly assayed is limited. Thus, it can be difficult to distinguish reliably some classes of relationship (e.g. full-sibs from half-sibs) with microsatellite data alone. We assess, via Monte Carlo computer simulation, the potential for using a large panel of independently segregating SNPs to infer genetic relationships, following the analytical approach of Blouin et al. (1996). We have explored a 'best case scenario' in which 100 independently segregating SNPs are available. For discrimination among single-generation relationships or for the identification of parent-offspring pairs, it appears that such a panel of moderately polymorphic SNPs (minor allele frequency of 0.20) will provide discrimination power equivalent to only 16-20 independently segregating microsatellites. Although newly available analytical methods that can account for tight genetic linkage between markers will, in theory, allow improved estimation of relationships using thousands of SNPs in highly dense genomic scans, in practice such studies will only be feasible in a handful of model organisms. Given the comparable amount of effort required for the development of both types of markers, it seems that microsatellites will remain the marker of choice for relationship estimation in nonmodel organisms, at least for the foreseeable future. PMID- 12753223 TI - Wolbachia infection complexity among insects in the tropical rice-field community. AB - Wolbachia are a group of intracellular bacteria that cause reproductive alterations in their arthropod hosts. Widely discordant host and Wolbachia phylogenies indicate that horizontal transmission of these bacteria among species sometimes occurs. A likely means of horizontal transfer is through the feeding relations of organisms within communities. Feeding interactions among insects within the rice-field insect community have been well documented in the past. Here, we present the results of a polymerase chain reaction-based survey and phylogenetic analysis of Wolbachia strains in the rice-field insect community of Thailand. Our field survey indicated that 49 of 209 (23.4%) rice-field insect species were infected with Wolbachia. Of the 49 infected species, 27 were members of two feeding complexes: (i) a group of 13 hoppers preyed on by 2 mirid species and parasitized by a fly species, and (ii) 2 lepidopteran pests parasitized by 9 wasp species. Wolbachia strains found in three hoppers, Recilia dorsalis, Nephotettix malayanus and Nisia nervosa, the two mirid predators, Cyrtorhinus lividipennis and Tytthus chinensis, and the fly parasitoid, Tomosvaryella subvirescens, were all in the same Wolbachia clade. In the second complex, the two lepidopteran pests, Cnaphalocrocis medinalis and Scirpophaga incertulas, were both infected with Wolbachia from the same clade, as was the parasitoid Tropobracon schoenobii. However, none of the other infected parasitoid species in this feeding complex was infected by Wolbachia from this clade. Mean (+/- SD) genetic distance of Wolbachia wsp sequences among interacting species pairs of the hopper feeding complex (0.118 +/- 0.091 nucleotide sequence differences), but not for the other two complexes, was significantly smaller than that between noninteracting species pairs (0.162 +/- 0.079 nucleotide sequence differences). Our results suggest that some feeding complexes, such as the hopper complex described here, could be an important means by which Wolbachia spreads among species within arthropod communities. PMID- 12753224 TI - Side-stepping secondary symbionts: widespread horizontal transfer across and beyond the Aphidoidea. AB - To elucidate the co-evolutionary relationships between phloem-feeding insects and their secondary, or facultative, bacterial symbionts, we explore the distributions of three such microbes--provisionally named the R-type (or PASS, or S-sym), T-type (or PABS), and U-type--across a number of aphid and psyllid hosts through the use of diagnostic molecular screening techniques and DNA sequencing. Although typically maternally transmitted, phylogenetic and pairwise divergence analyses reveal that these bacteria have been independently acquired by a variety of unrelated insect hosts, indicating that horizontal transfer has helped to shape their distributions. Based on the high genetic similarity between symbionts in different hosts, we argue that transfer events have occurred recently on an evolutionary timescale. In several instances, however, closely related symbionts associate with related hosts, suggesting that horizontal transfer between distant relatives may be rarer than transmission between close relatives. Our findings on the prevalence of these symbionts within many aphid taxa, along with published observations concerning their effects on host fitness, imply a significant role of facultative symbiosis in aphid ecology and evolution. PMID- 12753225 TI - Effects of transgenic Bt corn litter on the earthworm Lumbricus terrestris. AB - A 200-day study was carried out to investigate the impact of transgenic Bacillus thuringiensis (Bt) corn on immature and adult Lumbricus terrestris in the field and in the laboratory. Another objective of this study was to develop test methods that could be used for standard testing of the impact of transgenic plants on different earthworm species in the field and in the laboratory. For this purpose two different experiments were involved, a laboratory experiment with adult L. terrestris and a field experiment with immature L. terrestris. No lethal effects of transgenic Bt corn on immature and adult earthworms were observed. Immature L. terrestris in the field had a very similar growth pattern when fed either (Bt+) or (Bt-) corn litter. No significant differences in relative weights of (Bt+) and (Bt-) corn-fed adult L. terrestris were observed during the first 160 days of the laboratory trial, but after 200 days adult L. terrestris had a significant weight loss of 18% of their initial weight when fed (Bt+) corn litter compared to a weight gain of 4% of the initial weight of (Bt-) corn-fed earthworms. Further studies are necessary to see whether or not this difference in relative weight was due to the Bt toxin or other factors discussed in the study. Degradation of Cry1Ab toxin in corn residues was significantly slower in the field than at 10 degrees C in the laboratory. Enzyme-linked immunosorbent assay results indicated that earthworms in both experiments were exposed to the Bt toxin throughout the whole experimental time. PMID- 12753226 TI - Patterns of nuclear DNA degeneration over time--a case study in historic teeth samples. AB - The amount of nuclear DNA extracted from teeth of 279 individual red fox Vulpes vulpes collected over a period spanning the last three decades was determined by quantitative polymerase chain reaction (PCR). Although teeth were autoclaved during initial collection, 73.8% of extracts contained sufficient DNA concentration (> 5 pg/ micro L) suitable for reliable microsatellite genotyping but the quantity of nuclear DNA decayed significantly over time in a nonlinear pattern. The success of PCR amplification across four examined canine microsatellites over time was dependent on fragment size. By including data from two different tests for human contamination and from frequencies of allelic dropout and false alleles, the methodological constraints of population genetic studies using microsatellite loci amplified from historic DNA are discussed. PMID- 12753227 TI - Low and homogeneous copy number of plasmid-borne symbiont genes affecting host nutrition in Buchnera aphidicola of the aphid Uroleucon ambrosiae. AB - The bacterial endosymbiont of aphids, Buchnera aphidicola, often provides amino acids to its hosts. Plasmid amplification of leucine (leuABCD) and tryptophan (trpEG) biosynthesis genes may be a mechanism by which some Buchnera over-produce these nutrients. We used quantitative polymerase chain reaction to assess the leuABCD/trpEG copy variability within Uroleucon ambrosiae, an aphid with a wide diet breadth and range. Both leuABCD and trpEG abundances are: (i) similar for aphids across 15 populations, and (ii) low compared to Buchnera from other aphid species (particularly trpEG). Consequently, the plasmid location of trpEG combined with Buchnera's chromosomal polyploidy may functionally limit, rather than increase, tryptophan production within Uroleucon ambrosiae. PMID- 12753228 TI - Matrilineal genetic structure within and among populations of the cooperatively breeding common marmoset, Callithrix jacchus. AB - Common marmosets are members of the family Callitrichidae, South American primates characterized by highly social group living and cooperative breeding. In this study we analysed 1112 base pairs (bp) of the mitochondrial control region in 59 Callithrix jacchus individuals, sampled mainly from two geographically distinct field sites in N.E. Brazil. Analysis of molecular variation revealed a highly significant genetic structuring of haplotypes between social groups and between populations. Examination of matrilineal genetic structure within social groups revealed that seven of nine recorded breeding pairs were from different maternal lineages, indicating assortative mating and outbreeding. In addition to the breeders, at least six of 10 groups contained adult individuals from different matrilines, with five haplotypes present in one social group of nine animals. Groups of mixed lineages raise questions about potential reproductive conflicts of interest, and the extent of kin-selected altruism in the evolution and maintenance of cooperative breeding in this species. PMID- 12753239 TI - Production of dyestuffs from indole derivatives by naphthalene dioxygenase and toluene dioxygenase. AB - AIMS: To isolate and characterize the phorate [O,O-diethyl-S-(ethylthio)methyl phosphoradiothioate] degrading bacteria from agricultural soil, and their assessment for multifarious biological activities of environmental and agronomic significance. METHODS AND RESULTS: Based on their morphological and biochemical characteristics, the selected isolates PS-1, PS-2 and PS-3 were presumptively identified as Rhizobium, Pseudomonas and Proteus species, respectively. The HPLC analysis of phorate in bioaugmented soil revealed its complete disappearance within 40 days. The degradation isotherms of the isolates PS-1, PS-2 and PS-3 suggested time-dependent disappearance of phorate following the first-order rate kinetics at the corresponding rate constants of 0.04, 0.05 and 0.04 d-1. Besides, the isolates concurrently exhibited substantial phosphate solubilization, indole acetic acid (IAA) and siderophore production, as well as limited biocontrol activity against fungal phytopathogens. CONCLUSIONS, SIGNIFICANCE AND IMPACT OF THE STUDY: The data revealed the potential of isolates for collateral plant growth promotion, biocontrol and bioremediation. The selected strains may serve as an important bioresource for development of effective super-bioinoculants. PMID- 12753240 TI - Isolation and characterization of phorate degrading soil bacteria of environmental and agronomic significance. AB - Phorate [O,O-diethyl-S-(ethylthio)methyl phosphoradiothioate] degrading bacteria were isolated from agricultural soil and characterized based on their morphological and biochemical characteristics. The selected isolates PS-1, PS-2 and PS-3 were presumptively identified as Rhizobium, Pseudomonas and Proteous species, respectively. The HPLC analysis of phorate in bioaugmented soil revealed its complete disappearance within 40 days. The degradation isotherms of the isolates PS-1, PS-2 and PS-3 suggested time-dependent disappearance of phorate following the first order rate kinetics at the corresponding rate constants of 0.04, 0.05 and 0.04 days-1. Besides, the isolates concurrently exhibited substantial phosphate solubilization, indole acetic acid, and siderophore production. The isolate PS-3 also showed anti-fungal activity against a phytopathogen Fusarium oxysporum. As a result of the multifarious biological properties, the isolates have been suggested to be important bioresource for efficient bioinoculant development. PMID- 12753241 TI - Methane oxidation and its coupled electron-sink reactions in ruminal fluid. AB - AIMS: This study was conducted to investigate the occurrence of methane oxidation in the rumen, and to identify the electron-sink reaction coupled to the oxidation if it occurred. METHODS AND RESULTS: Mixed ruminal microbes taken from sheep were incubated with 13CH4. Oxidation of methane, estimated from the flux of 13C to CO2 and microbial cells, occurred, but represented only 0.2-0.5% of the methane produced. Methane oxidation was suppressed by the presence of oxygen, and was also inhibited by 2-bromoethane-sulphonate, and molybdate, but not by tungstate. CONCLUSION, SIGNIFICANCE AND IMPACT OF THE STUDY: Methane could be oxidized anaerobically in the rumen by reverse methanogenesis in consort with sulphate reduction. PMID- 12753242 TI - Improving heterologous gene expression in Aspergillus niger by introducing multiple copies of protein-binding sequence containing CCAAT to the promoter. AB - AIMS: To increase the expression level of heterologous gene in Aspergillus niger. METHODS AND RESULTS: Inserting multiple copies of an activator protein-binding site originally encoded in the cis-regulatory region of A. niger glucoamylase gene (glaA), CCAAT-containing sequence, into the promoter of an expression plasmid greatly enhanced the production of heterologous protein. CONCLUSION: The promoter activity was increased by the introduction of the CCAAT-containing sequence resulting in increased gene expression. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides evidence that the CCAAT-containing region of the glaA promoter of A. niger is a binding site for a positive transcriptional factor and demonstrates the possibility for improving the expression of both heterologous and endogenous genes in Aspergillus. PMID- 12753243 TI - Evaluation of biotracers to monitor effluent retention time in constructed wetlands. AB - AIMS: With concern surrounding the environmental impact of chemical tracers on the aquatic environment, this paper presents the initial evaluation of biotracers used to determine the effluent retention time, an important performance indicator, in a Free Water Surface Constructed Wetland. METHODS AND RESULTS: Production of the biotracers, coliphage MS2, and the bacteriophage of Enterobacter cloacae and antibiotic resistant endospores of Bacillus globigii is described in detail. Their subsequent use in three separate tracer experiments - January, March and June (2000) - revealed the variability of retention time with respect to effluent flow. The biotracer MS2 showed the constructed wetland had a retention time of 8-9 h at a mean discharge of 0.9 l s-1, increasing to 10-12 h at a mean discharge 0.3 l s-1. A similar retention of 9-10 h at a mean discharge of 0.3 l s-1 was calculated for the Ent. cloacae phage. In contrast, use of endospores revealed considerably longer retention times at these mean discharge rates; 12-24 h and 36-48 h, respectively. CONCLUSION: Biotracers could provide a useful and environmentally friendly technique to monitor effluent retention in constructed wetlands. At this stage the phage tracers appear particularly promising due to ease of isolation and recovery. SIGNIFICANCE AND IMPACT OF THE STUDY: Initial results are encouraging and have highlighted the potential of biotracers as alternatives to chemical tracers, even in microbially-rich waters. PMID- 12753244 TI - Detection of antibodies to Staphylococcus aureus Toxic Shock Syndrome Toxin-1 using a competitive agglutination inhibition assay. AB - AIMS: To develop a competitive agglutination inhibition assay (CAIA) for the detection of anti-Toxic Shock Syndrome Toxin-1 (TSST-1) antibody in serum samples using a commercially available reverse passive agglutination assay (RPLA) kit. METHODS AND RESULTS: TSST-1 toxin and sera were incubated together, so that anti toxin IgG would complex with the toxin. Latex particles sensitized with rabbit IgG anti-TSST-1 were added to test for un-complexed toxin. The sensitivity and specificity of the CAIA assay was determined relative to positive and negative ELISA results. The sensitivity (proportion of positive ELISA sera which tested positive by CAIA) was 66% whilst the specificity (proportion of ELISA negative sera which tested negative by CAIA) was 75%. Seven sera (14%) were negative by ELISA but positive for CAIA and 12 (18.8%) were positive for ELISA but negative for CAIA, suggesting some interference with the assays. Statistical analysis showed no significant difference between the methods in terms of the numbers of individuals testing positive (chi2, P = 0.04). CONCLUSIONS: The CAIA assay allowed detection of anti-TSST-1 within 18 h and was simple to read visually. SIGNIFICANCE AND IMPACT OF THE STUDY: The method is a useful test for individual serum samples and a preliminary investigation for medical screening of suspected toxic shock syndrome and is applicable in situations where antibody assays are not routinely used for anti-TSST-1 and also where sophisticated equipment (e.g. microtitre plate reader) is not available. PMID- 12753245 TI - Effect of phenolic monomers on biomass and hydrolytic enzyme activities of an anaerobic fungus isolated from wild nil gai (Baselophus tragocamelus). AB - AIMS: To test the anaerobic fungus, Piromyces sp. FNG5, for its tolerance to phenolic monomers released in the rumen by degradation of lignocellulosic poor quality feeds. METHODS AND RESULTS: Effects of phenolic monomers on biomass and fibrolytic enzyme activities of a pure culture of lignocellulolytic anaerobic fungus (Piromyces sp. FNG5) isolated from faeces of wild nil gai (blue bull, Baselophus tragocamelus) were evaluated. There was a reduction in fungal biomass at 1 mm concentration of catechol with complete inhibition at 10 mm. p-Coumaric acid caused a reduction in biomass at 10 mm and no growth was observed above 20 mm concentration. The fungal isolate could tolerate up to 5 mm of ferulic acid without any reduction in biomass level, and was able to grow to some extent up to the highest level of ferulic acid tested (20 mm). Vanillic acid had no effect on biomass of the fungus even up to 50 mm level. The phenolic monomers varied in their potential to inhibit the secretion of carboxymethyl cellulase, xylanase, beta-glucosidase and acetyl esterase activities with catechol being the most inhibitory and vanillic acid being the least inhibitory. After 14 days of incubation, 38.49-65.14%p-Coumaric acid, 65.22-74.10% ferulic acid and 34.13 66.78% vanillic acid disappeared from the medium under anaerobic conditions. CONCLUSIONS, SIGNIFICANCE AND IMPACT OF THE STUDY: It is concluded that the anaerobic fungus Piromyces sp. FNG5 is tolerant to phenolic monomers and has ability to degrade them. Therefore, such anaerobic fungi may play an important role in fibre degradation in the rumen. PMID- 12753246 TI - Influence of W-7, a calmodulin antagonist on phospholipid biosynthesis in Candida albicans. AB - AIM: This study was undertaken to investigate the role of calmodulin in phospholipid biosynthesis in Candida albicans using W-7, a calmodulin antagonist. METHODS: Cells were grown as shake cultures in the absence and presence of W-7 at different concentrations. Changes in cell mass, phospholipid content and incorporation of labelled precursor into phospholipid and activities of respective enzymes have been studied. RESULTS: Decreased incorporation of labelled acetate into total lipids and phospholipids was observed in the presence of 40 microm of W-7 which was not as a consequence of altered growth of Candida in the presence of calmodulin antagonist. Further, a significant decrease in the levels of calmodulin and CaM dependent protein kinase activity was observed in cells grown with different concentrations of W-7. This was accompanied by decreased/increased activity of phosphatidic acid phosphatase and phospholipase A, respectively in W-7 grown cells as compared to controls. CONCLUSIONS: These findings suggest definite involvement of calmodulin in the regulation of phospholipid metabolism in Candida albicans. PMID- 12753247 TI - A bleaching herbicidal activity of methoxyhygromycin (MHM) produced by an actinomycete strain Streptomyces sp. 8E-12. AB - AIMS: To screen bioherbicidal isolates and evaluate herbicidal activity of methoxyhygromycin (MHM) produced by Streptomyces sp. 8E-12. METHODS AND RESULTS: Streptomyces sp. 8E-12 with herbicidal activity was selected through seed germination bioassay. An active metabolite, MHM was isolated from culture broth by carbon absorption, butanol extraction, silica gel, Sephadex LH-20 and G-10 chromatography, and preparative HPLC. The metabolite was identified by electrospray ionization mass spectra (ESI-MS) and 1H- and 13C-NMR spectral data analyses. In vivo herbicidal activity was examined against weeds and crops grown on pots. CONCLUSIONS: Streptomyces sp. 8E-12 produced a selective herbicidal metabolite which was identified as MHM. The metabolite showed stronger in vivo activity against monocotyledonous plants than dicotyledonous plants, and caused a bleaching (albino symptom) on some weeds including Digitaria sanguinalis and Echinochloa crus-galli. SIGNIFICANCE AND IMPACT OF THE STUDY: These results showed that Streptomyces sp. 8E-12 produced a bioherbicidal metabolite, MHM and can be developed as a biocontrol agent (BCA) for weed control. PMID- 12753248 TI - The influence of mineral and carbon sources on biological control of charcoal rot fungus, Macrophomina phaseolina by fluorescent pseudomonads in tomato. AB - AIMS: To determine the influence of various trace minerals and carbon source on the biocontrol performance of Pseudomonas aeruginosa strain IE-6S+ and P. fluorescens strain CHA0 against Macrophomina phaseolina. METHODS AND RESULTS: In dual culture plate assay, P. aeruginosa IE-6S+ and P. fluorescens CHA0 inhibited radial growth of M. phaseolina producing zones of inhibition. Czapek's dox agar medium amended with both zinc and glucose remarkably improved antifungal activities of the bacterial inoculants. Under glasshouse conditions, soil amendment with zinc and/or glucose alone did not reduce M. phaseolina infection in tomato roots but did reduce significantly when used in combination with IE-6S+ or CHA0. Soil amendments with zinc and/or glucose increased fresh shoot weights but zinc amendment greatly reduced bacterial populations in the rhizosphere. CONCLUSIONS: Mineral and carbon amendments enhance the biocontrol performance of fluorescent pseudomonads against M. phaseolina. SIGNIFICANCE AND IMPACT OF THE STUDY: Identification of mineral and carbon amendments that favour biocontrol of certain bacterial strains may provide clues to soil factors or components of nutrient solutions in hydroponic culture that will improve the level and reliability of control. PMID- 12753249 TI - Regulation of branched-chain amino acid biosynthesis by alpha-acetolactate decarboxylase in Streptococcus thermophilus. AB - AIMS: To demonstrate the presence of an active alpha-acetolactate decarboxylase in Streptococcus thermophilus and to investigate its physiological function. METHODS AND RESULTS: Streptococcus thermophilus CNRZ385 contains a gene encoding an alpha-acetolactate decarboxylase. Comparison of the production of alpha acetolactate and its decarboxylation products, by the parent strain and an alpha acetolactate decarboxylase-deficient mutant, demonstrated the presence of a control of the pool of alpha-acetolactate by valine, leucine and isoleucine. This control occurs via an allosteric activation of the alpha-acetolactate decarboxylase. Cell-free extracts of S. thermophilus were not able to decarboxylate the isoleucine precursor alpha-acetohydroxybutyrate. CONCLUSIONS: These results strongly suggest that one of the physiological functions of the alpha-acetolactate decarboxylase in S. thermophilus is to regulate leucine and valine biosynthesis by diverting the flux of alpha-acetolactate towards acetoin when the branched-chain amino acids are present at a high concentration. SIGNIFICANCE AND IMPACT OF THE STUDY: Regulation of branched-chain amino acid biosynthesis by alpha-acetolactate decarboxylase may occur in several other micro organisms and explain some of their growth properties. PMID- 12753250 TI - Abattoir sources of psychrophilic clostridia causing blown pack spoilage of vacuum-packed chilled meats determined by culture-based and molecular detection procedures. AB - AIMS: To identify the abattoir source(s) of psychrophilic clostridia causing 'blown pack' spoilage of vacuum-packed chilled meats. METHODS AND RESULTS: Molecular procedures were used to detect the presence of specific 16S rRNA gene fragments of blown pack-causing clostridia in samples collected from a commercial abattoir and its environs. Blown pack-causing clostridia were consistently detected in hide, soil and faecal samples, as well as in samples collected at slaughter plant locations associated with handling of animals and animal carcasses prior to pelt removal. CONCLUSIONS: The data indicate that pelts per se or soil particles/faecal material attached thereto are the most probable primary reservoir of blown pack clostridia in the abattoir. SIGNIFICANCE AND IMPACT OF THE STUDY: The paper provides information critical for controlling blown pack spoilage in commercial meat-processing plants. PMID- 12753251 TI - A sensitive and rapid method to determine the viability of freeze-dried bacterial cells. AB - AIMS: To determine the potential use of flow cytometry for viability asssessment of freeze-dried bacterial cells. METHODS AND RESULTS: Escherichia coli CIP 54.8T and Vibrio metschnikovii CIP 104262 were analysed. The viability of freeze-dried cells resuspended in a nutrient broth was evaluated by culture whereas activity was determined by flow cytometry analysis of both esterase activity and cell death. Activity assessment by flow cytometry was found to be a rapid and good indicator of cell viability and was very efficient for quality control. For V. metschnikovii the fraction of active cells varies greatly depending on the freeze drying procedure and within a given procedure. CONCLUSIONS, SIGNIFICANCE AND IMPACT OF THE STUDY: Bacterial activity assessment by flow cytometry is very efficient for the control of freeze-dried cells. PMID- 12753252 TI - Molecular detection of anthrax spores on animal fibres. AB - AIMS: To develop a rapid, specific and sensitive diagnostic test for the detection of the spores of Bacillus anthracis on commercial samples of animal fibres (e.g. wool and cashmere). METHODS AND RESULTS: Extraction of DNA from spores using a mechanical disruption method based on bead beating was evaluated but subsequently abandoned as it compromised the sensitivity of the overall protocol. A multiplex PCR and two nested amplification reactions designed for B. anthracis were developed during this study. CONCLUSIONS: A simple selective incubation step in combination with multiplex PCR was found to be more effective than generic DNA extraction coupled to a sensitive nested amplification reaction. SIGNIFICANCE AND IMPACT OF THE STUDY: The rapid diagnostic test could be applied to the analysis of commercial fibre samples for the detection of anthrax as required by health and safety legislation resulting in considerable savings in time and expense. PMID- 12753255 TI - Regulation of bone remodeling and emerging breakthrough drugs for osteoporosis and osteolytic bone metastases. AB - Major advances have been made in the past 10 years in our understanding of the molecular basis of bone cell formation and bone remodeling. For example, the discovery of osteoprotegerin, the decoy receptor and inhibitor of receptor activator of NF-kappaB ligand (RANKL), and the RANKL/receptor activator of NF kappaB (RANK) signaling pathway that is essential for osteoclastogenesis, has helped clarify the mechanisms regulating osteoclast formation, activation, and survival. PTH, like most other osteoclast stimulating factors, promotes RANKL production by osteoblast/stromal cells when they are exposed to it continuously, but when given intermittently it stimulates bone formation and reduces fracture risk in postmenopausal women. This anabolic effect is associated with increased expression of insulin-like and fibroblast growth factors and decreased osteoblast apoptosis. Src tyrosine kinase is essential for osteoclast activation and also negatively regulates osteoblast activity. Thus, it is a well-validated therapeutic target for the prevention of postmenopausal and other forms of bone loss. Preliminary in vitro and in vivo studies of specifically designed, bone targeted, non-peptide Src inhibitors have shown that these compounds inhibit bone resorption and stimulate new bone formation. The design of drugs using structure/function approaches such as this should lead to the development of novel therapeutics that could be used to counteract the negative effects of chronic renal failure on the skeleton. PMID- 12753253 TI - Application of polymerase chain reaction for detection of Vibrio parahaemolyticus associated with tropical seafoods and coastal environment. AB - AIMS: To study the incidence of Vibrio parahaemolyticus in seafoods, water and sediment by molecular techniques vs conventional microbiological methods. METHODS AND RESULTS: Of 86 samples analysed, 28 recorded positive for V. parahaemolyticus by conventional microbiological method, while 53 were positive by the toxR targeted PCR, performed directly on enrichment broth lysates. While one sample of molluscan shellfish was positive for tdh gene, trh gene was detected in three enrichment broths of molluscan shellfish. CONCLUSIONS: Direct application of PCR to enrichment broths will be useful for the rapid and sensitive detection of potentially pathogenic strains of V. parahemolyticus in seafoods. SIGNIFICANCE AND IMPACT OF THE STUDY: Vibrio parahaemolyticus is an important human pathogen responsible for food-borne gastroenteritis world-wide. As, both pathogenic and non-pathogenic strains of V. parahaemolyticus exist in the seafood, application of PCR specific for the virulence genes (tdh & trh) will help in detection of pathogenic strains of V. parahaemolyticus and consequently reduce the risk of food-borne illness. PMID- 12753256 TI - Vitamin D receptor: mechanisms for vitamin D resistance in renal failure. AB - 1,25-dihydroxyvitamin D [1,25(OH)2D3], the hormonal form of vitamin D, controls serum levels of parathyroid hormone (PTH) and parathyroid hyperplasia. Both 1,25(OH)2D3 actions involve regulation of gene transcription by the 1,25(OH)2D3/vitamin D receptor (VDR) complex. In advanced renal failure, in addition to low serum 1,25(OH)2D3 and reduced parathyroid vitamin D receptor content, several mechanisms downstream from 1,25(OH)2D3/VDR complex formation contribute to the impairment of 1,25(OH)2D3 action, including reduced levels of the retinoid X receptor, RXR, with the consequent reduction in VDR/RXR heterodimer formation, and accumulation of uremic toxins and increases in nuclear levels of calreticulin, two processes that impair the binding of the VDR/RXR complex to vitamin D responsive elements in vitamin D-regulated genes. VDR/RXR heterodimer formation and its binding to DNA is critical for 1,25(OH)2D3 regulation of gene transcription. Early interventions with 1,25(OH)2D3 could delay the onset of vitamin D resistance by preventing both 1,25(OH)2D3 deficiency and its critical consequence, reduction in VDR content. Once established, vitamin D resistance could be counteracted by vitamin D analogs. While their less calcemic properties make higher dosing safer, their specificity to recruit co activator molecules to the transcriptional pre-initiation complex could compensate for reduced 1,25(OH)2D3/VDR by potentiating VDR transactivation/transrepression of genes critical for normal PTH synthesis and parathyroid cell growth. PMID- 12753257 TI - Importance of arachidonic acid as a mediator of parathyroid gland response. AB - The intracellular signaling mechanisms that mediate the regulation of parathyroid hormone (PTH) secretion by parathyroid glands are becoming increasingly more understood. Extracellular calcium modulates parathyroid function by acting on a G protein-coupled calcium-sensing receptor, which activates the hydrolysis of membrane phospholipids by phospholipases C, D, and A2 to generate intracellular signals. Arachidonic acid (AA) produced by phospholiphase A2 (PLA2) appears to play a crucial role throughout the generation of downstream-oxygenated products. Recent studies demonstrate the activation of the PLA2 via an intracellular calcium increase, and that the elevation of cytosolic calcium also overcomes the repressive effect of high extracellular phosphate on AA production. Furthermore, a role of the mitogen-activated protein (MAP) kinase cascade has also been documented in PLA2 activation. PMID- 12753258 TI - Influence of polymorphisms in VDR and COLIA1 genes on the risk of osteoporotic fractures in aged men. AB - BACKGROUND: Osteoporosis in chronic renal failure is a common finding caused by several factors, including age. In the last decade, the likely effect of genetic markers related with the appearance and evolution of osteoporosis has been mainly studied in women, with no categorical results. The aim of this study was to assess the influence of polymorphisms of the vitamin D receptor (VDR) and COLIA1 genes on the risk of osteoporotic fractures in men older than 50 years. METHODS: The study population comprised 156 men, aged 64 +/- 9 (50-86), randomly selected from the population list of Oviedo, Spain. Prevalent vertebral fractures and incident non-vertebral fractures were identified, as well as several genetic polymorphisms. Prevalent vertebral fractures were considered according to the Genant grade 2 classifications. The analyzed genetic polymorphisms were located on restriction sites BsmI (B,b), ApaI (A,a), and TaqI (T,t) in the VDR and on Sp1 (S,s) in COLIA1. RESULTS: Although none of the VDR gene polymorphisms separately analyzed showed any differences between fractured and non-fractured men, the utilization of haplotypes could be employed in order to find osteoporotic fractures in men. By contrast, the COLIA1 polymorphism was associated with osteoporotic fractures. The percentage of prevalent vertebral fractures was significantly higher in the "ss" genotype with respect to the other genotypes. These results show that in men, the "ss" genotype of COLIA1 polymorphism could be the best osteoporotic fracture risk genetic predictor, independent of bone mass values. PMID- 12753259 TI - Response of parathyroid glands to calcitriol in culture: Is this response mediated by the genetic polymorphisms in vitamin D receptor? AB - BACKGROUND: Recently, we have developed a model of parathyroid tissue culture that allows the study of the response of the parathyroid glands to long-term effectors, such as calcitriol, and that is also useful to study the likely effect of the genetic polymorphisms in the functionality of the glands. The aim of this study was to evaluate the response to calcitriol of cultured parathyroid tissue from patients with secondary hyperparathyroidism (HPT) and the possible effect of vitamin D receptor (VDR) gene polymorphisms on this response. METHODS: Parathyroid glands (N = 37) from 34 parathyroidectomized patients (17 men, 17 women) were used. Several gland fragments were cultured for 60 hours in the presence of calcitriol 10(-9) mol/L or 10(-8) mol/L. DNA from each fragment was extracted to normalize the hormone secretion levels and to genotype the restriction sites ApaI, BsmI, TaqI, and FokI in the VDR gene. RESULTS: The percentages of secretion observed in the response to calcitriol were: 69%+/- 28% (range, 3-100) and 46%+/- 19% (range, 8-78) for calcitriol 10(-9) mol/L and 10( 8) mol/L, respectively (P = 0.004). None of the polymorphisms showed statistical differences in response to calcitriol with any of the concentrations used. CONCLUSION: Parathyroid glands cultured in vitro from patients with secondary HPT are able to respond to calcitriol decreasing PTH synthesis. These results, however, do not support the current hypothesis that VDR polymorphisms are involved in the modulation of the parathyroid gland response. PMID- 12753260 TI - Effect of VDR gene polymorphisms on osteocalcin secretion in calcitriol stimulated human osteoblasts. AB - BACKGROUND: The impact of vitamin D receptor (VDR) gene polymorphisms in bone metabolism remains controversial. Some authors have found a beneficial effect of some VDR gene polymorphisms, while others found no differences, or even a lower bone mass in subjects with the same type of polymorphisms. The aim of this study was to assess if the VDR gene polymorphisms could have an effect on the calcitriol-stimulated osteocalcin in human osteoblasts. METHODS: Osteoblasts were obtained from human femoral necks replaced because of osteoarthritis. Bones were cut into pieces of 1 to 2 mm and placed in a nylon mesh. After the migration of osteoblasts, the pieces were collected and cultured with different concentrations of calcitriol (10(-8), 10(-9), and 10(-1)0 mol/L). After 48 hours of incubation with calcitriol, the osteocalcin secreted into the medium (corrected by either total proteins or total DNA content) was measured. The DNA was extracted from the osteoblasts, amplified by polymerase chain reaction (PCR), and analyzed for target sequences sites of the BsmI, ApaI, TaqI, and FokI restriction enzymes. RESULTS: The response observed in osteocalcin secretion in the bb or TT genotypes doubled the response observed in the BB or tt genotypes (calcitriol 10(-8) and 10(-9) mol/L). A slight trend was also observed with the aa genotype. Men showed higher levels of osteocalcin secretion than women. Age did not show any influence in osteocalcin secretion. CONCLUSION: VDR alleles and gender demonstrated an effect on the osteocalcin secretion. BB or tt genotypes, and also the "A" allele, showed the lowest calcitriol-stimulated osteocalcin secretion. PMID- 12753261 TI - Comparative study of the expression of proteins involved in the cell cycle in renal secondary hyperparathyroidism. AB - BACKGROUND: In renal hyperparathyroidism, parathyroid cell proliferation seems to play a key role in the progression of the disease. Therefore, G1/S transition, a main cell cycle regulatory step, could be deregulated in these patients. METHODS: One hundred and one parathyroid glands, taken from parathyroidectomies performed on 41 patients on hemodialysis (HD), and 15 glands, taken from 7 patients with post-transplantation persistent hyperparathyroidism (HPT), were studied. Twelve normal parathyroid (PT) glands were used as the control. Biochemical data, immunohistochemical (IHC) profiles of G1/S transition regulators belonging to the two main pathways (cyclin D1/p16INK4A/pRb and p14ARF/p53/MDM2), and proliferation rate (Ki67) were correlated. RESULTS: All of the other proteins differed from normal IHC profiles in both groups that showed significant higher proliferating rates, decreases in p27KIP1, pRb, and cyclin D1, as well as increases in p16INK4A, p53, MDM2, and p21WAF1 levels, in comparison with normal PT glands, with the exception of cyclin D3. Contrary to patients with HPT who were on hemodialysis, in post-transplantation HPT, consistent correlations between biochemical data and IHC profiles were obtained. CONCLUSION: In both groups IHC profiles of proteins involved in G1/S transition regulation significantly differed from normal PT glands. The results support partial reversion to normal IHC profile in post-transplantation HPT. PMID- 12753262 TI - Chromosomal aberrations, the consequence of refractory hyperparathyroidism: its relationship with biochemical parameters. AB - BACKGROUND: It has been shown that refractory hyperparathyroidism (HPT) correlates biologically with a monoclonal true neoplasm, but the chromosomal changes and their relationship with biochemical variables such as high levels of phosphate, low levels of calcium (Ca), and calcitriol deficiency are still in need of a deeper analysis. METHODS: Comparative genomic hybridization was used to scan for DNA copy number changes in two groups of samples: 57 glands from refractory secondary HPT and 28 glands from refractory HPT after kidney transplantation. Biochemical HPT-related parameters from these patients were collected and analyzed. RESULTS: Sixty-one percent of the glands from dialysis patients and 53.6% of the glands from transplanted patients suffering severe secondary hyperparathyroidism had clonal chromosomal imbalances. Losses were far more common than gains. The most recurrent changes were losses of 1p (71%), monosomies of chromosomes 19 and 22 (45%), and losses of 20q (44%) and 16p (42%). The most frequent gains were 5q, 6q, and 13q. Biochemical parameters suggested that Ca excess is related to the development of these chromosomal aberrations, although it is not known if it is by playing a role in producing the alterations or merely as a reflection of HPT severity. Phosphate levels, despite their known effect in increasing the proliferation of the parathyroid glands, were not related to the chromosomal aberrations found in severe secondary HPT. CONCLUSION: Clonal recurrent chromosomal changes are present in more than half of the glands from patients with refractory HPT, which undergo extreme biochemical levels in hyperparathyroidism effectors. These changes support the idea of the monoclonal neoplastic nature of this disorder. PMID- 12753263 TI - Effect of aluminium on calcium-sensing receptor expression, proliferation, and apoptosis of parathyroid glands from rats with chronic renal failure. AB - BACKGROUND: To assess the effect of aluminium on the calcium-sensing receptor expression, proliferation, and apoptosis in parathyroid glands from rats with chronic renal failure, 2(1/2)-month-old male Wistar rats were 7/8 nephrectomized. METHODS: Eight weeks after surgery the rats were divided into two groups, one receiving intraperitoneal AlCl3 for 8 weeks and the other receiving intraperitoneal placebo. Serum Al, Ca, P, creatinine, and PTH were measured. Parathyroid glands were removed, formaldehyde-fixed, and paraffin-embedded. Calcium-sensing receptor and proliferation were detected by immunohistochemistry and apoptosis by TUNEL and propidium iodide uptake. RESULTS: At the end of the study, despite higher levels of serum P in the aluminium group (6.27 +/- 0.63 vs. 5.56 +/- 0.58 mg/dL; P = 0.045), serum PTH was lower (89.6 +/- 57.7 vs. 183.1 +/- 123.8 pg/mL; P = 0.059). No significant differences were found in the calcium sensing receptor expression between groups (aluminium: 27.1 +/- 7.6; placebo: 25.4 +/- 3.5 RU). Rats receiving aluminium showed a significantly lower cell proliferation rate than the control rats (0.54 +/- 0.69 vs. 4.43 +/- 3.10 cells/mm2; P = 0.003). No apoptotic events were detected. CONCLUSION: Aluminium was able to reduce the cell proliferation of the parathyroid glands. Due to the low apoptosis rate, however, it was not possible to find any change. Aluminium had no effect on the calcium-sensing receptor expression. PMID- 12753264 TI - Vitamin D status and secondary hyperparathyroidism: the importance of 25 hydroxyvitamin D cut-off levels. AB - BACKGROUND: Serum 25-hydroxyvitamin D is the best indicator of vitamin D status. However, some controversy remains regarding "normal" and "abnormal" values. This study's aim was to assess vitamin D status and prevalence of secondary hyperparathyroidism. METHODS: A random sample of 326 subjects (164 women and 162 men, aged 68 +/- 9; range, 54 to 89) participating in the European Vertebral Osteoporosis Study (EVOS) was used to assess vitamin D status and secondary hyperparathyroidism. Only those subjects who had never received any kind of treatment for osteoporosis were included in this analysis. RESULTS: Serum 25 hydroxyvitamin D levels were "deficient" (<10 ng/mL) in 27% of subjects, "borderline" (10-18 ng/mL) in 40% of subjects, and "normal" (>18 ng/mL) in 33% of subjects. The prevalence of secondary hyperparathyroidism (PTH>65 pg/mL) according to 25-hydroxyvitamin D levels was 33% (<10 ng/mL), 16% (10-18 ng/mL), and 12% (>18 ng/mL), respectively. There were no cases of secondary hyperparathyroidism with 25-hydroxyvitamin D levels>40 ng/mL. The independent predictors for PTH were 25-hydroxyvitamin D and serum creatinine in both sexes, but age was a predictor only in men. CONCLUSION: These remarkable findings demonstrate the importance of maintaining higher 25-hydroxyvitamin D levels to avoid stimulation of the parathyroid gland. PMID- 12753265 TI - Use of active vitamin D sterols in patients with chronic kidney disease, stages 3 and 4. AB - This paper reviews randomized controlled trials and other reported data on the use of the active vitamin D sterols, such as calcitriol, alfacalcidol, and doxercalciferol, in the management of secondary hyperparathyroidism in patients with mild-to-moderate renal insufficiency (stage 3 or 4 chronic kidney disease). Data on potential benefits, including improved histologic abnormalities of bone from secondary hyperparathyroidism, increased bone mineral density, and a reduction of elevated parathyroid hormone levels, are documented. Consideration is given to the risks of such therapy, which include the production of hypercalcemia, more rapid progression of renal insufficiency, the induction of adynamic bone "disease," and accelerated vascular and soft tissue calcification. The low therapeutic index, or "benefit/risk ratio" of calcitriol and alfacalcidol, the sterols currently licensed for such treatment, is recognized. It is recommended that phosphate-restricted diets, phosphate-binding agents, and oral calcium supplements be given adequate trials before starting calcitriol or alfacalcidol. If PTH levels cannot be controlled by these measures, initial doses of these sterols and the proper surveillance during treatment are given. It is emphasized that the risks of hypercalcemia are likely to increase as the degree of kidney failure worsens. Further research using the "less calcemic" vitamin D sterols is clearly needed. PMID- 12753266 TI - The renal osteodystrophy pattern in Brazil and Uruguay: an overview. AB - BACKGROUND: The prevalence of the types of renal osteodystrophy (ROD) has changed in the last two decades. This study evaluates the prevalence and determining factors of the types of ROD in two countries in South America. METHODS: Symptomatic patients who underwent bone biopsies for diagnostic purposes (1985 through 2001) were included in the study. In Brazil, a total of 2340 patients were reviewed, 93.1% on hemodialysis (HD), and 6.9% on continuous ambulatory peritoneal dialysis (CAPD). In Uruguay, 167 patients on hemodialysis were included. Uncalcified bone samples were analyzed in order to diagnose the different types of ROD. RESULTS: Comparing the 1980s to the 1990s, the prevalence of Brazilian patients with hyperparathyroid bone disease (HP) increased from 32.3% to 44.0%, while Al overload decreased from 61.3% to 42.4%. A logistic regression analysis showed that the main factors associated with HP included female gender, age of less than 40 years, black patients, longer time on dialysis, and absence of Al overload. In Uruguay, HP also increased over time from 31.8% to 70.5% (P < 0.05), while Al overload decreased from 42% to 27% (P < 0.05). CONCLUSION: A better control of Al overload may lead to a change in histologic pattern, as evidenced by the increasing prevalence of HP. PMID- 12753267 TI - Hormonal replacement therapy in an animal model with chronic renal failure and ovariectomy: biochemical and densitometric study. AB - BACKGROUND: In spite of estrogen replacement therapy being extensively used in clinical and experimental studies without renal impairment, there are no long term studies concerning estrogen replacement in chronic renal failure. METHODS: In this experimental study, six groups of nephrectomized and ovariectomized animals were treated with different doses of 17beta-estradiol, alone or in combination with calcitriol, to evaluate the effect of these treatments on bone metabolism. RESULTS: Biochemical results showed that estrogen alone did not have any effect neither on calcium nor on PTH serum levels. By contrast, in the groups treated with calcitriol, the levels of serum calcium were significantly higher, and the levels of iPTH were significantly lower than those observed in the control group. Animals receiving the combined treatment with estrogen and calcitriol showed the greater gain in uterus weight and a better bone mineral density at the lumbar site and the proximal and distal tibia sites. CONCLUSION: The combination of estrogen and calcitriol is the most effective therapy to prevent bone mass loss in animals with chronic renal failure and estrogen deprivation. PMID- 12753268 TI - Selective estrogen receptor modulators in chronic renal failure. AB - BACKGROUND: In addition to renal osteodystrophy, postmenopausal women on dialysis could be at risk of osteoporosis. Hormone replacement therapy (HRT) could have beneficial effects as well as potentially serious risks, especially in uremic women, due to the pharmacokinetics of estradiol in renal failure. Therapeutic alternatives, such as the selective estrogen receptor modulators (SERMs), have shown the benefits of estrogen on bone and serum lipid levels, without its adverse effects on the breast and endometrium, in nonuremic women. METHODS: Recent data on the effect of the SERM raloxifene in bone and lipid metabolism in osteoporotic postmenopausal women on dialysis is reviewed. Since the estrogen receptor (ER) gene has been suggested as a candidate marker for osteoporosis, we investigated whether ER polymorphism could have predicted the BMD response to raloxifene. RESULTS: Hemodialyzed women on raloxifene demonstrated increased trabecular bone mineral density (BMD) and decreased bone resorption markers. Similarly, LDL-cholesterol values dropped significantly. ER gene polymorphism analysis of baseline BMD parameters did not differ between PP/xx or Pp/Xx groups. Nevertheless, patients on raloxifene with PP/xx genotypes, but not those with Pp/Xx, showed a higher trabecular BMD after one year on treatment, suggesting that homozygous women for P or x alleles of the ER have a better BMD response to raloxifene. CONCLUSION: Raloxifene and, most likely, other SERMs, could represent a good alternative to HRT in postmenopausal uremic women. PMID- 12753269 TI - Evolution of serum phosphate in long intermittent hemodialysis. AB - BACKGROUND: Phosphate (P) control is crucial to prevent secondary hyperparathyroidism and extraosseous calcification. Hemodialysis techniques normally used do not remove the amount of P absorbed from an adequate protein intake. Our long hemodialysis (LHD) procedure produces greater P removal in a seven-hour session than in a conventional four-hour session. Our aim was to assess the evolution of predialysis serum P and Ca x P product in patients on intermittent LHD during the first two years on this dialysis schedule. METHODS: Eight irregular compliant patients underwent LHD for 12 months; five of them completing 24 months. In the conventional hemodialysis (CHD) period, session time was 12 hours per week, while it was 21 hours per week in LHD. We compared P and Ca x P product throughout both periods; each patient was his or her own control. RESULTS: Mean serum P improved from 7.1 +/- 1.1 mg/dL in CHD to 5.6 +/- 1.1 mg/dL during the second year on LHD (P < 0.05). Mean Ca x P product decreased from 64 +/- 12.7 mg2/dL2 during CHD to 50.9 +/- 10.3 mg2/dL2 at the end of the second year of LHD. CONCLUSION: By prolonging dialysis time to 21 hours per week, better P serum control and lower Ca x P product were achieved in our patients. PMID- 12753270 TI - Treatment of hyperphosphatemia with sevelamer hydrochloride in hemodialysis patients: a comparison with calcium acetate. AB - BACKGROUND: Sevelamer hydrochloride is a recently approved calcium- and aluminium free phosphate binder. A randomized study comparing sevelamer and calcium acetate was performed to assess the control of hyperphosphatemia in hemodialysis patients. METHODS: Administration of phosphate binders was discontinued during a two-week washout period. The patients were then randomized to receive sevelamer or calcium acetate. The laboratory tests were performed monthly for 34 weeks. RESULTS: There was a statistically significant decrease of serum phosphorus in both sevelamer and calcium acetate treatments. In addition, sevelamer improved the lipid profile. CONCLUSION: This study confirms that sevelamer is effective at lowering serum phosphorus in hemodialysis patients and that it has several striking properties that could be beneficial in atherosclerosis in dialysis patients. PMID- 12753271 TI - A multicenter study on the effects of lanthanum carbonate (Fosrenol) and calcium carbonate on renal bone disease in dialysis patients. AB - BACKGROUND: Lanthanum carbonate (LC) (Fosrenol) is a novel new treatment for hyperphosphatemia. In this phase III, open-label study, we compared the effects of LC and calcium carbonate (CC) on the evolution of renal osteodystrophy (ROD) in dialysis patients. METHODS: Ninety-eight patients were randomized to LC (N = 49) or CC (N = 49). Bone biopsies were taken at baseline and after one year of treatment. Acceptable paired biopsies were available for static and dynamic histomorphometry studies in 33 LC and 30 CC patients. Blood samples were taken at regular intervals for biochemical analysis and adverse events were monitored. RESULTS: LC was well tolerated and serum phosphate levels were well controlled in both treatment groups. The incidence of hypercalcemia was lower in the LC group (6% vs. 49% for CC). At baseline, subtypes of ROD were similarly distributed in both groups, with mixed ROD being most common. At one-year follow-up in the LC group, 5 of 7 patients with baseline low bone turnover (either adynamic bone or osteomalacia), and 4 of 5 patients with baseline hyperparathyroidism, had evolved toward a normalization of their bone turnover. Only one lanthanum-treated patient evolved toward adynamic bone compared with 6 patients in the CC group. In the LC group, the number of patients having either adynamic bone, osteomalacia, or hyperpara decreased overall from 12 (36%) at baseline to 6 (18%), while in the calcium group, the number of patients with these types of ROD increased from 13 (43%) to 16 (53%). CONCLUSION: LC is a poorly absorbed, well-tolerated, and efficient phosphate binder. LC-treated dialysis patients show almost no evolution toward low bone turnover over one year (unlike CC-treated patients), nor do they experience any aluminum-like effects on bone. PMID- 12753272 TI - Advantages of adjusting the initial dose of intravenous calcitriol according to PTH levels. AB - BACKGROUND: To assess the usefulness of starting calcitriol therapy with a dose proportional to the degree of hyperparathyroidism, 141 patients from 28 centers were treated with intravenous calcitriol for 6 months. The aim was to achieve a final PTH between 125 and 250 pg/mL. Patients with serum PTH>250 pg/mL were included in the study and divided into 4 groups according to baseline PTH levels. METHODS: The study was completed by 100 patients, a third of which were treated strictly according to the protocol, labeled "compliants"; thus, calcitriol was started according to baseline PTH levels. Two thirds of patients, labeled "noncompliants," showed one or more violation in the dosage regimen. RESULTS: After 2 months of treatment with calcitriol, 59% of the "compliants" and 35% of the "noncompliants" decreased their PTH levels>40% (P = 0.022), 70%, and 49%, respectively after 3 months of treatment. After 3 months of treatment, 67% of the "compliants" reached the target (PTH 125 to 250 pg/mL) in contrast with 23% of the "noncompliants" (P < 0.001). The number of hypercalcemic and hyperphosphatemic episodes was significantly lower in the "compliants" group (P < 0.006). CONCLUSION: These results demonstrate several advantages when calcitriol therapy is started with a dose proportional to the severity of hyperparathyroidism. PMID- 12753274 TI - Usefulness of pamidronate in severe secondary hyperparathyroidism in patients undergoing hemodialysis. AB - BACKGROUND: Although bisphosphonates have been widely used to treat bone diseases characterized by increased bone resorption, there are limited data showing their possible usefulness in patients on hemodialysis (HD) with secondary hyperparathyroidism. METHODS: The aim of this study was to evaluate the efficacy and safety of pamidronate in HD patients affected by severe secondary hyperparathyroidism and moderate hypercalcemia who were receiving intravenous calcitriol (Calcijex). RESULTS: In this prospective one-year, open-labeled study, 13 patients (9 women/4 men) with a mean age of 64 +/- 9 years and a mean time on dialysis of 94 +/- 61 months were evaluated. The inclusion criteria were: iPTH>500 pg/mL, Ca>11 mg/dL, P <6 mg/dL, and osteopenia (T-score <-1 SD). Blood levels of Ca, P, alkaline phosphatase (AP), and iPTH were assessed at the beginning of the study and every month. Radiographs of the vertebral spine and bone mineral density (BMD) (lumbar spine and femoral neck) were assessed basal and every 6 months. All patients received 60 mg of pamidronate intravenously every two months throughout the study period. Calcitriol and phosphate binders were adjusted according to iPTH, Ca, and P blood levels. BMD increased in both the lumbar and femoral neck scans (mean increase of 33%) at 6 and 12 months. iPTH increased at 3 months in all patients, and decreased more than 50% in 10 patients after increasing the calcitriol doses. Three patients had no response. A slight decrease in Ca and P was observed in all patients with no significant changes in AP. There were no adverse events. CONCLUSION: Pamidronate is effective in controlling hypercalcemia in patients on HD with secondary hyperparathyroidism and allows for a more aggressive use of intravenous calcitriol. PMID- 12753273 TI - New vitamin D analogs. AB - BACKGROUND: 1,25-(OH)2D3 (calcitriol) controls parathyroid gland growth and suppresses the synthesis and secretion of parathyroid hormone. Because of this, 1,25-(OH)2D3 has been used successfully for the treatment of secondary hyperparathyroidism, which almost always accompanies renal failure. However, the potent effect of 1,25-(OH)2D3 on intestinal calcium and phosphorus absorption and bone mineral mobilization often leads to the development of hypercalcemia and hyperphosphatemia precluding 1,25-(OH)2D3 therapy. METHODS: This has led to the development of vitamin D analogs that retain the suppressive action on PTH and parathyroid gland growth, but that have less calcemic and phosphatemic activity. Currently, two analogs, 19-nor-1,25-(OH)2D2 and 1,alpha(OH)D2, are being used for the treatment of secondary hyperparathyroidism in the United States, and two are being used in Japan, 22-oxa-calcitriol and 1,25-(OH)2-26,27F6 D3. RESULTS: All four analogs suppressed PTH, but had less calcemic and phosphatemic activity than 1,25-(OH)2D3. In rats, 19-nor-1,25-(OH)2D2 has been shown to be less calcemic and phosphatemic compared to 1,alpha(OH)D2. CONCLUSION: Therapeutic doses of 19-nor 1,25-(OH)2D2 could produce a lower Ca x P product compared to 1,alpha(OH)D2, which could be an important consideration in patient treatment. Further studies are necessary to define these differences and to understand the mechanisms behind the differential actions of vitamin D analogs. PMID- 12753275 TI - Calcimimetic agents: review and perspectives. AB - BACKGROUND: Recognition of the role of the extracellular calcium-sensing receptor (CaR) in mineral metabolism has greatly improved our understanding of calcium homeostasis. The activation of this receptor by small changes in the extracellular ionized calcium (ec(Ca2+)) regulates PTH, calcitonin secretion, urinary calcium excretion, and, ultimately, bone turnover. METHODS: The cloning of the CaR and the discovery of mutations that make the receptor less or more sensitive to calcium have allowed a better understanding of several hereditary disorders characterized by either hyperparathyroidism or hypoparathyroidism. The CaR, able to amplify the sensitivity of the CaR to Ca++ and suppress PTH levels with a resulting decrease in blood Ca++, became an ideal target for the development of compounds, the calcimimetics. Experience with the calcimimetic R 568 in patients with primary and secondary hyperparathyroidism and parathyroid carcinoma are summarized. RESULTS: The first clinical studies with the first generation calcimimetic agents have demonstrated their efficacy in lowering plasma intact PTH concentration in uremic patients with secondary hyperparathyroidism. However, the low bioavailability of these first calcimimetics predicts a difficult clinical utilization. The second-generation calcimimetic, AMG 073, having a better pharmacokinetic profile, appears to be effective and safe for the treatment of secondary hyperparathyroidism, suppressing PTH levels while simultaneously reducing serum phosphorus levels and the calcium x phosphorus product. CONCLUSION: The advantage of controlling PTH secretion without the complications related to hypercalcemia, hyperphosphatemia, and increased calcium x phosphorus product is very promising. PMID- 12753276 TI - Parathyroidectomy: whom and when? AB - Hyperparathyroidism (HPT) is common in patients on dialysis, and parathyroidectomy (PTx) is often required. We present a retrospective, descriptive analysis of data corresponding to 148 patients on dialysis undergoing PTx due to severe refractory HPT (PTH 1401 +/- 497 pg/mL, Ca 10.6 +/- 0.8 mg/dL, P 6.9 +/- 1.7 mg/dL). Demographic data were compared with those recorded in 309 patients on dialysis not subjected to PTx who were managed at the same hospital. In the PTx group, the factors age (49.3 +/- 14 years), male gender (48.6%), and diabetes (0.7%) were significantly lower than in the non-PTx group (61.5 +/- 14.9 years, male gender 59%, diabetes 19.4%), while time on dialysis was longer (8.6 +/- 5.8 vs. 5.5 +/- 5.4 years). In 129 of the study patients (87.4%), four or more glands were identified, and total PTx plus autotransplantation (AT) in the forearm was performed. In the remaining 19 patients, two to three glands were identified, and AT was not undertaken. Four of the 19 patients were successfully operated on again for persistent HPT, seven showed PTH levels <250 pg/mL, and eight maintained severe HPT. Perioperative complications included one death due to cardiac insufficiency, two repeat operations due to bleeding, and one patient with chronic hoarseness. Hospital stay was prolonged in 20% of patients due to a hungry bone syndrome. Among those patients with PTx and AT, HPT recurred in 21 patients (16.2%) at 3.1 +/- 2.3 years. In 13 of these patients, autograft was removed at 7.5 +/- 2.9 years. Serum calcium and phosphate levels improved after PTx, and these results were maintained for 5 years (9.6 +/- 0.8 and 4.2 +/- 1.2 mg/dL, respectively). In conclusion, PTx with AT is a safe option for the treatment of severe HPT that is accompanied by low morbidity and mortality and a good outcome. Medical treatment should not be prolonged at the expense of long repeated bouts of hypercalcemia and/or hyperphosphatemia with their irreversible consequences. PMID- 12753277 TI - Treatment of refractory secondary hyperparathyroidism with ethanol injection: the importance of glandular volume. AB - BACKGROUND: Percutaneous ethanol injection treatment (PEIT) has been proposed as an alternative to surgery for patients with secondary hyperparathyroidism. The present study was undertaken to determine factors that may predict results. METHODS: We performed PEIT in 19 patients with secondary hyperparathyroidism refractory to medical therapy under ultrasonographic guidance in an ambulatory facility with local anesthesia. Biochemical assays were performed immediately before the last dialysis session (basal) and between 1 to 7 days after PEIT (post PEIT). RESULTS: Serum PTH, calcium, and phosphorus levels decreased significantly after treatment. The percent of change in serum PTH was significantly correlated to total nodular volume (r = 0.73, P = 0.0004), and basal PTH levels (r = 0.48, P = 0.03). Post-PEIT serum phosphate and calcium x phosphate product disclosed negative correlations that were statistically significant with the decrease of PTH levels (r = -0.60, P = 0.009, and r = -0.60, P = 0.01, respectively). The total nodular volume was significantly correlated to the percent change in serum calcium levels (r = 0.60, P = 0.01), in phosphate levels (r = 0.64, P = 0.009), and calcium x phosphate product (r = 0.66, P = 0.01). CONCLUSION: Our findings suggest that patients with uncontrolled secondary hyperparathyroidism may benefit from PEIT if they present with very high basal PTH levels and/or big nodule size. PMID- 12753278 TI - Traditional and emerging cardiovascular risk factors in end-stage renal disease. AB - Patients with end-stage renal disease face a particularly high risk of cardiovascular disease and total mortality. Part of their increased risk is due to a higher prevalence of established risk factors, such as arterial hypertension, diabetes, smoking, and anemia. Hypertension and diabetes have a very high prevalence in dialysis patients and play a major role in their high mortality and morbidity. Hyperparathyroidism, hyperhomocysteinemia and disordered lipid metabolism represent factors that are peculiarly altered by the uremic state. Inflammatory processes, high sympathetic activity, and the accumulation of an endogenous inhibitor of NO synthase (ADMA), have recently emerged as cardiovascular risk factors of paramount importance. Sleep apnea has been linked with nocturnal hypertension and could be implicated in the high prevalence of concentric hypertrophy of the left ventricle in these patients. Hypertension control, as well as appropriate treatment of anemia and cessation of smoking, constitutes a fundamental area of intervention in dialysis patients. It appears possible that, in the near future, control of chronic inflammatory processes of high sympathetic activity and endothelial dysfunction will further help to curb the exceedingly high cardiovascular mortality of patients on chronic dialysis treatment. PMID- 12753279 TI - Higher impact of mineral metabolism on cardiovascular mortality in a European hemodialysis population. AB - BACKGROUND: Cardiovascular disease is the main cause of morbidity and mortality among hemodialysis patients. Chronic renal failure influences a number of factors that cause accelerated atherogenesis, with calcium, phosphorus, and PTH playing key roles. Several studies have demonstrated the influence of these factors on all-cause and cardiovascular mortality in the American hemodialysis population. In the present study we evaluated the variables that influence long-term cardiovascular mortality in a European hemodialysis population. METHODS: One hundred and forty-three hemodialysis patients were followed for six years. Several Cox models were used to study the influence of demographic and biochemical data, and comorbid conditions in cardiovascular survival, with a particular interest in mineral metabolism. RESULTS: There was an increased risk of cardiovascular death in patients with serum P>6.5 mg/dL (risk ratio [RR], 2.5), PTH>50 pmol/L (RR, 3.9), Ca x P>52 (RR, 2.8), BB or Bb genotype (RR, 3.8), and in diabetics. CONCLUSION: There is a stronger influence of mineral metabolism on cardiovascular death among European patients when compared to the American population. PMID- 12753280 TI - Heart valve calcification and calcium x phosphorus product in hemodialysis patients: analysis of optimum values for its prevention. AB - BACKGROUND: Prevalence of valve calcification (VC) in end-stage renal disease (ESRD) patients is high and information regarding modifiable predictors is scarce. Our aim was to determine the prevalence of VC in our maintenance hemodialysis (HD) population, and the optimal Ca x P value that most accurately predicted the presence of VC after controlling for comorbidities. METHODS: This was a cross-sectional observational study of a cohort of 52 stable patients on maintenance HD for more than 12 months. Mean 12 months serum biochemical data (calcium, phosphorus, PTH, lipids) and M-mode 2D echocardiogram were used to evaluate the presence or absence of mitral and aortic VC and ventricular geometry. RESULTS: Twenty patients (38.4%) presented with VC. Patients with VC were more commonly diabetic and showed higher levels of serum phosphorus, Ca x P product, total and LDL cholesterol, and poor ventricular geometry, as compared to patients without VC. Moreover, they required higher doses of both CaCO3 and Al(OH)3. Logistic regression analysis showed that VC was independently influenced by age, Ca x P, and diabetes. ROC curves illustrated that a Ca x P>43 mg2/dL2 was the optimal value in terms of sensitivity and specificity for predicting the presence of VC in our patient population. CONCLUSION: These findings highlight the importance of applying more vigorous measures for Ca x P control. PMID- 12753281 TI - Vascular calcification in the uremic patient: a cardiovascular risk? AB - BACKGROUND: Several factors suggest that the presence of vascular calcification (VC) is associated with a high risk of cardiac events in uremic patients. The aim of this study was to analyze the influence of VC on cardiac morbidity and mortality in our hemodialysis (HD) patients. METHODS: We studied 79 patients on HD: 43 males, mean age 48 +/- 15 years old, mean time on HD 83 +/- 63 months. The presence of VC was evaluated by radiologic series. Other cardiovascular risk factors analyzed were arterial hypertension, diabetes mellitus, obesity, cigarette smoking, anemia, and dyslipidemia. All patients underwent M-mode, two dimensional, Doppler echocardiography. Patients were followed for two years. During this time, clinical information collected included predialysis blood pressure, incidence of ischemic heart disease, episodes of congestive heart failure, and mortality due to cardiovascular event. RESULTS: VC was observed in 55.7% of patients. Left ventricular hypertrophy, diastolic dysfunction, and cardiac valve calcification were significantly associated with VC. Ischemic heart disease (71.4% vs. 28.6%) and episodes of cardiac failure (0.41 vs. 0.18 per year; P < 0.05) appeared more frequently in the patient group with VC. VC was present in 80.6% of patients who developed episodes of heart failure. Eight patients died from cardiac disease; each of them had VC. CONCLUSION: The presence of VC can help to identify those HD patients with a higher cardiovascular risk. PMID- 12753282 TI - The evolving clinical features of calciphylaxis. PMID- 12753283 TI - Bone remodeling after renal transplantation. AB - Several studies have indicated that bone alterations after transplantation are heterogeneous. Short-term studies after transplantation have shown that many patients exhibit a pattern consistent with adynamic bone disease. In contrast, patients with long-term renal transplantation show a more heterogeneous picture. Thus, while adynamic bone disease has also been described in these patients, most studies show decreased bone formation and prolonged mineralization lag-time faced with persisting bone resorption, and even clear evidence of generalized or focal osteomalacia in many patients. Thus, the main alterations in bone remodeling are a decrease in bone formation and mineralization up against persistent bone resorption, suggesting defective osteoblast function, decreased osteoblastogenesis, or increased osteoblast death rates. Indeed, recent studies from our laboratory have demonstrated that there is an early decrease in osteoblast number and surfaces, as well as in reduced bone formation rate and delayed mineralization after transplantation. These alterations are associated with an early increase in osteoblast apoptosis that correlates with low levels of serum phosphorus. These changes were more frequently observed in patients with low turnover bone disease. In contrast, PTH seemed to preserve osteoblast survival. The mechanisms of hypophosphatemia in these patients appear to be independent of PTH, suggesting that other phosphaturic factors may play a role. However, further studies are needed to determine the nature of a phosphaturic factor and its relationship to the alterations of bone remodeling after transplantation. PMID- 12753284 TI - Bone disease in long-term adult kidney transplant patients with normal renal function. AB - BACKGROUND: In successful renal transplantation, the degree of renal function recovery is usually incomplete and information is scarce about the abnormalities of mineral metabolism in long-term adult renal recipients with normal renal function. This study was designed to investigate bone mineral metabolism in patients with a long-term normal functioning kidney. METHODS: Twenty-nine adult asymptomatic renal transplant (RT) recipients with stable graft function for more than 10 years and serum creatinine <2 mg/dL were studied. They were classified into two groups according to glomerular filtration rate: Group A (N = 12; nine men, three women)>70 mL/min (x: 126 +/- 55 mL/min) and Group B (N = 17; nine men, eight women) <70 mL/min (x: 56 +/- 11 mL/min). Circulating biochemical markers of bone remodelling, bone histomorphometry, and densitometry (lumbar spine and hip) were obtained to investigate bone disease in these patients. RESULTS: Serum PTH was slightly elevated in 10 patients (83%) in group A. Serum PTH levels were positively related to serum calcium, osteocalcin, BAP, telopeptide, OH-proline, and creatinine. There was no histologic data to support overactivity on bone in this group of patients, with only one showing high bone turnover. Mineralization was prolonged in 34% of patients. Twenty-two patients (75%) exhibited normal bone turnover. In the group with GFR>70 mL/min the prevalence of mineralization defect in the presence of normal serum levels of calcitriol suggested vitamin D resistance. Lumbar and femoral neck osteoporosis was present in 25% and 33% of patients in group A, and 23% and 53% in group B, respectively. T-score at lumbar spine was negatively correlated with months since transplantation. Patients under treatment with cyclosporine (CsA) showed increased concentrations of osteocalcin and D-pyr and higher lumbar bone mineral density (BMD), but bone histomorphometry was not influenced by CsA. CONCLUSION: Patients with long-term renal transplantation with normal renal function frequently present with slight increases in PTH, but without an effect on bone histology. CsA did not induce changes in bone histology and delayed mineralization was frequently observed. PMID- 12753285 TI - Cyclic AMP activates B-Raf and ERK in cyst epithelial cells from autosomal dominant polycystic kidneys. AB - BACKGROUND: The proliferation of mural epithelial cells is a major cause of progressive cyst enlargement in autosomal-dominant polycystic kidney disease (ADPKD). Adenosine 3', 5' cyclic monophosphate (cAMP) stimulates the proliferation of cells from ADPKD cysts, but not cells from normal human kidney cortex (HKC), through the activation of protein kinase A (PKA), mitogen-activated protein kinase kinase (MEK), and extracellular signal-regulated kinase (ERK/MAPK). In the current study, we examined the signaling pathway between PKA and MEK in ADPKD and HKC cells. METHODS: Primary cultures of human ADPKD and HKC cells were prepared from nephrectomy specimens. We determined the effects of cAMP and epidermal growth factor (EGF) on the activation of ERK, B-Raf and Raf-1 in ADPKD and HKC cells by immune kinase assay and Western blot. RESULTS: 8-Br-cAMP increased phosphorylated ERK (2.7- +/- 0.6-fold, N = 7), and B-Raf kinase activity (3.6- +/- 1.1-fold, N = 5) in cells from ADPKD kidneys; levels of phosphorylated Raf-1 were not changed. Inhibition of PKA by H89 strikingly decreased cAMP-stimulated phosphorylation of ERK and B-Raf, and MAPK inhibition by PD98059 blocked the effect of the nucleotide to activate ERK. By contrast, in HKC cells 8-Br-cAMP did not activate B-Raf and ERK. EGF stimulated the phosphorylation of ERK and Raf-1 in both ADPKD and HKC cells, but had no effect on B-Raf. 8-Br-cAMP and EGF conjointly increased ERK activation above that of either agonist alone in ADPKD cells, and this combined effect was abolished by PD98059, indicating that ERK was activated by EGF- and cAMP-responsive cascades that converge at MAPK. CONCLUSION: cAMP activates ERK and increases proliferation of ADPKD epithelial cells, but not cells from normal human kidney cortex, through the sequential phosphorylation of PKA, B-Raf and MAPK in a pathway separate from, but complementary to, the classical receptor tyrosine kinase cascade. Consequently, cAMP and EGF have great potential to accelerate the progressive enlargement of renal cysts. PMID- 12753286 TI - Mutation analysis and clinical implications of von Willebrand factor-cleaving protease deficiency. AB - BACKGROUND: The pentad of thrombocytopenia, hemolytic anemia, mild renal dysfunction, neurologic signs, and fever, classically characterizes the syndrome of thrombotic thrombocytopenic purpura (TTP). TTP usually occurs in adults as an acquired form but a congenital form in children has also been described. In the latter case, the initial presentation is often with neonatal jaundice and thrombocytopenia. The disorder may subsequently take a relapsing course. Deficiency of a recently identified novel metalloprotease, the von Willebrand factor (vWF)-cleaving protease, originating from mutations in the ADAMTS13 gene plays a major role in the development of TTP. METHODS: Blood for DNA analysis was collected from six unrelated TTP families, consisting of nine patients from four different countries, and was screened for mutations in the ADAMTS13 gene. This gene spans 29 exons encompassing approximately 37 kb. Conventional techniques of DNA extraction, polymerase chain reaction (PCR), and direct cycle sequencing were used. RESULTS: Eight novel ADAMTS13 mutations are presented. Half of the total number of mutant ADAMTS13 alleles are amino acid substitutions. The disease causing mutations are spread over the gene. The pathogenicity of the individual mutations is based upon their predicted effect on the ADAMTS13 protein and segregation in family members. Although most of the patients (seven out of nine) had symptoms during the neonatal period, they were in a remarkably good condition. Only one of the nine patients had a decreased glomerular filtration rate (GFR) with proteinuria and hematuria. Another patient had epileptic seizures. CONCLUSION: We confirm that deficiency of ADAMTS13 is a molecular mechanism responsible for familial TTP. An early diagnosis allows prophylactic treatment with fresh plasma infusions. PMID- 12753287 TI - Role of Smad4 on TGF-beta-induced extracellular matrix stimulation in mesangial cells. AB - BACKGROUND: The best characterized signaling pathway employed by transforming growth factor-beta (TGF-beta) is the Smad pathway; however, its role in matrix production in mesangial cells is unclear. We focused on Smad4, as Smad4 is essential for the activation of Smad-dependent target genes. METHODS: To investigate the function of Smad4 in extracellular matrix (ECM) production, we generated several stably transfected mesangial cell lines (MMC) that have a deletion in the linker region (Smad4 Delta M4: Delta 275-322) or have a deletion in MH1 of Smad4 (Smad4N4: Delta 1-136). The ECM genes, alpha1 type I collagen (COL1A1), plasminogen activator inhibitor-1 (PAI-1) and fibronectin (FN) were assessed in wild-type mesangial cells and stably transfected Smad4-DN cell lines in the absence and presence of TGF-beta. RESULTS: As compared to wild-type MMC that had a 10.8-fold stimulation of TGF-beta-induced p3TP-Lux activity, MMC stably transfected with Smad4 Delta M4 and Smad4N4 had only a 2.0-fold and 1.3 fold stimulation, respectively, indicating that they had dominant-negative effects on TGF-beta signaling. Basal and TGF-beta-induced COL1A1 expression in Smad4 dominant-negative cells were dramatically reduced to very low levels. The early (2 hours) TGF-beta-induced PAI-1 mRNA expression was inhibited; however, the sustained (24 to 48 hours) TGF-beta-induced expression was not affected in Smad4 dominant-negative cells. For FN, TGF-beta-induced expression was maintained in Smad4-dominant negative cells. CONCLUSION: These results indicate that Smad4 is essential for basal and TGF-beta-induced COL1A1 expression, and contributes to the early, but not sustained TGF-beta-induced PAI-1 expression in mesangial cells. However, TGF-beta-induced FN expression is independent of Smad4. In conclusion, Smad4 has a discriminate effect in mediating specific ECM molecules stimulated by TGF-beta in mesangial cells. PMID- 12753288 TI - Role of TGF-beta signaling in extracellular matrix production under high glucose conditions. AB - BACKGROUND: Hyperglycemia has been shown to play an important role in diabetic renal and vascular complications. Some studies show that high glucose may mediate diabetic complications by stimulating extracellular matrix (ECM) production. We hypothesize that this may be mediated by activating transforming growth factor beta (TGF-beta)/Smads signaling. METHODS: Renal and vascular cells were cultured under high glucose conditions in the presence or absence of a neutralizing TGF beta antibody and examined for activation of Smad signaling and collagen production. The regulating role of Smad signaling in high glucose-induced collagen synthesis was determined by inducing overexpression of the inhibitory Smad7 in a stable Smad7-expressing tubular cell line. RESULTS: Activation of Smad signaling, as evidenced by Smad2 and Smad3 nuclear translocation and phosphorylation, was found in renal and vascular cells at 24 hours after high glucose stimulation (up to 55% increased). This was associated with de novo synthesis of collagen I at day 3 by all cell types. High glucose-induced activation of Smad signaling and collagen synthesis were TGF-beta-dependent since these were associated with a significant increase in TGF-beta production at 24 hours (P < 0.01) and were blocked by a neutralizing TGF-beta antibody. Importantly, overexpression of Smad7 resulted in marked inhibition of high glucose-induced Smad2 and Smad3 activation and type I collagen synthesis, suggesting that Smad signaling is a key pathway in high glucose-mediated renal and vascular scarring. CONCLUSION: High glucose acts by activating the TGF-beta dependent Smad signaling pathway to stimulate collagen synthesis by renal and vascular cells. Smad signaling plays a critical role in regulating high-glucose mediated diabetic renal and vascular complications. PMID- 12753289 TI - Efficacy of 19-Nor-1,25-(OH)2D2 in the prevention and treatment of hyperparathyroid bone disease in experimental uremia. AB - BACKGROUND: The control of parathyroid hyperplasia and high circulating parathyroid hormone (PTH) levels is crucial in preventing secondary hyperparathyroidism (SH) in renal failure. Parathyroid gland enlargement and elevated levels of PTH are major contributors to increase bone resorption, a feature of renal osteodystrophy. METHODS: These studies assessed the efficacy of the 1,25(OH)2D3 analog, 19-Nor-1,25(OH)2D2 (19-Nor), in the prevention (protocol I) and treatment (protocol II) of SH and renal osteodystrophy in uremic rats. In protocol I, normal and uremic rats were fed a high phosphorus diet for 2 months; uremic rats were administered intraperitoneal injections of either vehicle or 19 Nor (200 ng three times a week). In protocol II, normal and uremic rats were fed a high phosphorus diet for 4 months; 2 months after the onset of uremia, rats were administered either intraperitoneal vehicle or 19-Nor (200 ng three times a week). Serum PTH and bone histology were used to assess the degree of SH. RESULTS: 19-Nor was effective in preventing (protocol I) and suppressing (protocol II) the significant SH induced by uremia and further enhanced by a high phosphorus diet. In protocol I, bone histology in uremic controls showed a threefold increase in the cancellous bone mass compared to normal rats. This expansion in unmineralized bone was accompanied by 5-, 1.5-, and 7-fold increases in eroded surface, mineralization lag time (MLT), and bone formation rate (BFR/BS), respectively. Moreover, cortical bone porosity in untreated uremic rats increased 267-fold compared to normal animals. 19-Nor ameliorated these changes in cancellous and cortical bone. In protocol II, the reported indices worsened even further. In contrast, 2 months of 19-Nor treatment improved bone histology by reducing cortical bone porosity, woven bone formation, MLT, and BFR/BS. CONCLUSION: In an experimental model of chronic renal failure (CRF), 19-Nor prevents SH and ameliorates the histomorphometric changes induced by uremia and high phosphorus diet. In addition, 19-Nor suppresses serum PTH and improves bone histology in uremic rats with established severe SH. Further studies in patients with CRF are necessary to define the clinical applicability of 19-Nor on bone histology in humans. PMID- 12753291 TI - Bone morphogenic protein-7 (BMP-7), a novel therapy for diabetic nephropathy. AB - BACKGROUND: Bone morphogenic protein-7 (BMP-7), an essential developmental renal morphogen, is a secreted differentiation factor of the adult collecting duct. It activates receptors in the collecting duct, distal nephron, proximal tubule, and glomerulus. BMP-7 is therapeutic in tubulointerstitial nephritis raising the question of broader efficacy in chronic kidney disease (CKD). METHODS: Diabetes was induced in 200 g rats by a single dose of streptozotocin. After 16 weeks, glomerular hypertrophy and proteinuria were established, and therapy with BMP-7 (10, 30, or 100 microg/kg intravenously twice a week), enalapril (20 mg/kg), or vehicle was begun and continued until 32 weeks. Kidney weight, glomerular filtration rate (GFR), urine albumin excretion, blood pressure, pathology, and BMP-7 expression were measured. RESULTS: Diabetic vehicle-treated rats developed renal insufficiency by 32 weeks (GFR, 0.34 +/- 0.02 mL/min/100 g body weight vs. 0.55 +/- 0.02 in normal). In the diabetic BMP-7 high-dose-treated rats, GFR was preserved (0.70 +/- 0.08, P < 0.01 vs. vehicle), and higher than diabetic enalapril-treated rats (0.58 +/- 0.06). Kidney weights of vehicle-treated animals were not affected, but were reduced in all of the treatment groups (P < 0.001). Proteinuria was reversed to normal by BMP-7 in a dose-dependent manner. The reduction in proteinuria by the intermediate dose of BMP-7 was similar to the effect of enalapril therapy. Glomerular area and interstitial volume were significantly decreased in the BMP-7 and enalapril-treated animals. Glomerular sclerosis was prevented by BMP-7 therapy more effectively than by enalapril. Enalapril controlled hypertension throughout the course of therapy while BMP-7 did not affect blood pressure until the final 4 weeks of therapy. Diabetic vehicle-treated rats lost BMP-7 expression in the kidney. BMP-7 and enalapril therapy restored BMP-7 expression at high levels. CONCLUSION: BMP-7 partially reversed diabetic-induced kidney hypertrophy, restoring GFR, urine albumin excretion, and glomerular histology toward normal. Restoration of BMP-7 expression was associated with a successful repair reaction and a reversal of the ill-fated injury response. PMID- 12753290 TI - Transforming growth factor-beta 1 increases glucocorticoid binding and signaling in macrophages through a Smad- and activated protein-1-mediated process. AB - BACKGROUND: Renal inflammation is regulated by a network of local and systemic mediators. Of them, transforming growth factor-beta1 (TGF-beta 1) and glucocorticoids play an important role in deactivating monocytes/macrophages. We examined the hypothesis that TGF-beta 1 effects may be partially achieved through modulation of the sensitivity of these cells to glucocorticoids. METHODS: Human promonocytic U 937 cells differentiated to a mature macrophage-like phenotype were exposed to recombinant TGF-beta 1 before specific binding of [3H] dexamethasone was measured. The expression of glucocorticoid receptor (GR) was examined by RNase protection assay and Western blot analysis. The role of Smad 2/3 and activator protein 1 (AP-1) in the response to TGF-beta 1 was determined by introducing transdominant negative mutants and decoy oligodeoxynucleotides, respectively. RESULTS: U 937 cell exposure to TGF-beta 1 caused a dose- and time dependent increase in [3H] dexamethasone binding to these cells, with a < or =twofold increase in the number of binding sites per cell, without modification of the affinity. The changes in glucocorticoid binding were associated with identical changes in GR protein and mRNA levels, that were explained by an increase in GR gene transcription rather than by posttranscriptional mechanisms. Functional inactivation of Smad 2/3 and AP-1 limited the response to TGF-beta 1, indicating a role for these transcription factors. Finally, increases in glucocorticoid binding to GR were responsible for increases in the ability of GR to transactivate minimal promoters containing glucocorticoid-responsive elements (GRE) [MMTV-Luc and (GRE)2 TK-Luc]. CONCLUSION: TGF-beta 1 increases glucocorticoid binding and signaling in inflammatory cells through a Smad 2/3- and AP-1-mediated process. This may represent a new target for intervention to increase glucocorticoid responsiveness. PMID- 12753292 TI - Effects of caspase inhibition on the progression of experimental glomerulonephritis. AB - BACKGROUND: Caspase-3 has a central role in the execution of apoptosis. In a nephrotoxic nephritis (NTN) model, we previously demonstrated an up-regulation of caspase-3 that was associated with inappropriate renal apoptosis, inflammation, tubular atrophy, and renal scarring. METHODS: We applied a pan caspase inhibitor, Boc-Asp (OMe)-fluoro-methyl-ketone (B-D-FMK), directly to rat NTN kidney using an intrarenal cannula fed from an osmotic pump. Animals were treated either for the first 7 days (acutely) to determine the effects on renal inflammation (ED-1 staining) and apoptosis (in situ end labeling of fragmented DNA), or for 28 days commencing 15 days after NTN (chronically) to observe the effects on cell death and renal fibrosis. Changes of caspase-3 and caspase-1 activity were detected by fluorometric substrate cleavage assay. Changes in caspase-3 and caspase-1, interleukin-1 beta (IL-1 beta), and collagen I, III, and IV proteins and mRNA were detected by Western blotting and Northern blotting, respectively. RESULTS: In both treated groups, caspase-3 activity was inhibited, and 17 and 24 kD active caspase-3 proteins were reduced significantly. A compensatory increase of caspase 3 mRNA occurred in the acutely treated group, but decreased in the chronically treated group (P < 0.05). Although there were no significant changes in caspase-1 activity and its active protein, the observed decrease in its precursor in the chronic group was increased by treatment (P < 0.05). Further, IL-1 beta precursor and its mRNA were significantly reduced by treatment only in the chronically treated group. Apoptosis was decreased in the glomeruli of acutely treated rats, and in the tubules and interstitium of chronically treated animals (P < 0.05). Glomerular inflammation was decreased only in the acutely treated group, whereas tubulointerstitial inflammation was lowered in both treated groups (P < 0.05). Glomerulosclerosis was reduced in both inhibitor groups, with a reduction in tubulointerstitial fibrosis and collagen I, III, and IV mRNA restricted to chronically treated animals (P < 0.05). Proteinuria was significantly decreased with caspase inhibition in both treated groups, but not serum creatinine level. CONCLUSION: This study clearly indicates that caspase inhibition reduces renal apoptosis, ameliorates inflammation and fibrosis, and improves proteinuria in experimental glomerulonephritis, which may mainly be related to changes in the caspase enzymatic system. PMID- 12753293 TI - Simvastatin suppresses tissue factor expression and increases fibrinolytic activity in tumor necrosis factor-alpha-activated human peritoneal mesothelial cells. AB - BACKGROUND: Patients treated with peritoneal dialysis frequently suffer from recurrent peritonitis episodes. During peritonitis, inflammatory mediators are released and a serofibrinous exudate is formed in the peritoneal cavity, which promotes fibrosis and abdominal adhesion development. Human peritoneal mesothelial cells (HMC) play a critical role in maintaining the intraperitoneal balance between fibrinolysis and coagulation by expressing the fibrinolytic enzyme tissue-type plasminogen activator (t-PA) and its specific inhibitor, plasminogen activator inhibitor-1 (PAI-1) as well as the procoagulant protein, tissue factor. METHODS: Cultured HMC were used to examine the effect of a 3 hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor, simvastatin, on the expression of t-PA, PAI-1 and tissue factor after activation of the cells with tumor necrosis factor-alpha (TNF-alpha). Antigen concentrations in the cell supernatants were measured by enzyme-linked immunosorbent assay (ELISA). Northern blot analysis was conducted for mRNA expression. Luciferase reporter gene assays and Western blot analysis in human fibrosarcoma HT1080 cells and HMC were performed to analyze the effect of simvastatin on the transcription factors nuclear factor kappa B (NF-kappa B) and activator protein-1 (AP-1), which regulate tissue factor gene expression. RESULTS: Incubation of HMC with TNF-alpha resulted in significantly decreased t-PA and increased PAI-1 synthesis. In the presence of simvastatin t-PA synthesis in control and TNF-alpha-treated cells dose-dependently increased, reaching 5.8-fold and 7.7-fold higher t-PA levels, respectively, at 5 micromol/L simvastatin after 48 hours. Simvastatin dose dependently suppressed PAI-1 production in both control and TNF-alpha-treated cells. At 5 micromol/L, simvastatin lowered PAI-1 synthesis 3.4-fold and 4.0 fold, respectively, thereby also completely suppressing the TNF-alpha effect itself. Similarly, simvastatin down-regulated the expression of tissue factor and also completely opposed the TNF-alpha-induced tissue factor expression. The effects of simvastatin on t-PA, PAI-1 and tissue factor expression were prevented by mevalonate and geranylgeraniol (GG), suggesting the involvement of geranylgeranyl-modified intermediates in simvastatin's mode of action. Also, simvastatin reduced NF-kappa B- and AP-1-dependent reporter gene activity in TNF alpha-treated HT-1080 fibrosarcoma cells and reduced the nuclear levels of p50-NF kappa B, p65-NF-kappa B, and the AP-1 components c-fos and c-jun in HMC. CONCLUSION: The HMG-CoA reductase inhibitor simvastatin is an effective stimulator of the mesothelial fibrinolytic capacity and suppresses the procoagulant activity both under normal and inflammatory conditions. Our findings provide a molecular explanation for the anti-inflammatory properties of statins in HMC and a rationale for the use of these drugs to protect peritoneal dialysis patients from peritoneal fibrosis and adhesion development during bacterial peritonitis. PMID- 12753294 TI - Conditionally immortalized cell line of inducible metanephric mesenchyme. AB - BACKGROUND: The mesenchymal-epithelial conversion of metanephric mesenchyme (MM) in the formation of nephronic tubules has long served as a paradigm for inductive signaling in morphogenesis. However, the mechanisms underlying this differentiation have remained an enigma due to insufficient numbers of primary mesenchymal cells that must be isolated manually from animal embryos. To overcome this problem, we have established a conditionally immortalized cell line, the rat inducible metanephric mesenchyme (RIMM-18) by transfection of primary mesenchymal cells with a vector, encoding an estradiol-dependent E1A-ER fusion protein. METHODS: Reverse transcription-polymerase chain reaction (RT-PCR), luciferase reporter assay, electrophoretic mobility shift assay, immunocytochemical, and immunohistochemical stainings were used to characterize the established cell line. RESULTS: We demonstrate that in the presence of estradiol, the RIMM-18 cell line proliferates continuously, maintaining mesenchymal characteristics for over 40 passages. These cells are vimentin-positive and cytokeratin-negative. Under inductive conditions in the absence of estradiol, they are responsive to a number of cytokines, which are established inducers of mesenchymal cells in vivo and in vitro [i.e., fibroblast growth factor 2 (FGF2), leukemia inhibitory factor (LIF), and transforming growth factor-beta 2 (TGF-beta 2)]. We show the presence in RIMM 18 cells of specific protein markers and functionally active signaling pathways required for induction of tubule formation in MM. Furthermore, induced RIMM-18 cells change morphology, acquiring epithelial-like features, and begin to express epithelial markers (e.g., E-cadherin, cytokeratin, gamma-glutamyl-transpeptidase, and secreted frizzled-related protein 2 (sFRP2). CONCLUSION: This preliminary characterization of the RIMM-18 cell line suggests that it will be useful in the study of biochemical and molecular mechanisms of nephronic development and, possibly, of some types of renal cancer such as Wilms' tumor, which caricatures the normal process of kidney development. PMID- 12753295 TI - Microvascular and tubulointerstitial injury associated with chronic hypoxia induced hypertension. AB - BACKGROUND: Rats submitted to chronic hypoxia develop hypertension that persists despite cessation of the hypoxia or correction of the hematocrit. We examined whether chronic hypoxia might induce subtle renal injury since studies in other animal models of hypertension suggest this may cause persistent hypertension. METHODS: Chronic hypoxia was induced in rats by placement in a hypobaric chamber for up to 24 days. Blood pressure and kidney biopsies were performed at baseline, 6 hours, 24 hours, and 24 days of hypoxia. RESULTS: Chronic hypoxia induced hypertension and erythrocytosis at 24 days. Acute hypoxia was associated with endothelial cell swelling in arterioles (6 and 24 hours), followed by thickening of the arterioles at 24 days. Subtle tubulointerstitial injury and inflammation occurred and was progressive. The influx of macrophages increased steadily over the 24 days and was associated with a progressive increase in interstitial collagen III deposition. Hypoxia was associated with increased tubular expression of osteopontin as early as 6 hours, the same period when an increase of proximal tubular cell proliferation occurred. Interstitial cell proliferation peaked twice, at 6 hours and at 24 days. Glomerular hypertrophy was manifest at 24 days. CONCLUSION: Both afferent arteriolar disease and subtle tubulointerstitial inflammation and injury occur early in hypoxic rats. These changes may predispose these animals to persistent hypertension. PMID- 12753296 TI - Epigenetic and genetic analysis of p16 in dermal fibroblasts from type 1 diabetic patients with nephropathy. AB - BACKGROUND: Several studies have shown that cultured skin fibroblasts from patients with diabetic nephropathy (DN) exhibit a hyperplastic growth phenotype. Increased DNA synthesis in cells from patients with DN may ultimately involve alterations in cell cycle regulatory proteins. p16 protein is a member of INK4 family of cyclin-dependent kinase inhibitors, which plays an important role in cell cycle regulation. In this study, we examined the correlation between p16 protein expression in cultured dermal fibroblasts from type 1 diabetic patients and the presence of DN. METHOD: Western blot analysis was performed to compare p16 protein expression in skin fibroblasts from patients with DN as compared to control subjects, diabetic patients without DN, and nondiabetic patients with nephropathy. Transcriptional regulation of the p16 gene was assessed using competitive reverse transcription-polymerase chain reaction (RT-PCR). Methylation status of the promoter region of p16 was examined using methylation-specific PCR, and we used single-stranded conformational polymorphism (SSCP)-PCR to assess p16 single-nucleotide polymorphism. RESULTS: Cells from diabetic patients with DN had nondetectable to significantly lower protein expression of p16. Similarly, mRNA expression of p16 was significantly lower in diabetic patients with DN. No hypermethylation of p16 gene was detected, and no abnormal migrating bands were noticed on SSCP-PCR analysis in cells from patients with DN. CONCLUSION: Our data indicate that cells from patients with DN exhibit significantly lower protein and mRNA expression of p16. This study could have not only important implications for the understanding of the pathogenesis of DN, but also the absence of p16 may ultimately serve as an early marker for DN. PMID- 12753297 TI - Correction of anemia in uremic mice by genetically modified peritoneal mesothelial cells. AB - BACKGROUND: During peritoneal dialysis, mesothelial cells become detached from the peritoneum and accumulate in the dialysate. Our aim was to evaluate the potential of peritoneal effluent (PF)-derived human peritoneal mesothelial cells (HPMC) as target for gene therapy. We used erythropoietin (EPO) as our target gene. METHODS: Various extracellular matrixes (ECM) were tested for optimal adhesion and growth of HPMC. The EPO gene was introduced to mouse peritoneal mesothelial cells (MPMC) and HPMC by transfection or retroviral transduction. EPO secretion from PMC was measured by enzyme-linked immunosorbent assay (ELISA) and by the TF-1 cell proliferation assay. We performed intraperitoneal or intramuscular transplantations of the genetically modified cells into regular or 5/6 nephrectomized Balb/c mice and nude mice. Finally, we measured serum EPO and hematocrit levels. RESULTS: ECM-coated plates provided up to sixfold increase in the efficiency of PMC isolation from PF. Gelatin coated dishes (20 microg/cm2) were found optimal for isolation of PF-HPMC. RPR-120535 liposome was found to be best for PMC transduction. In vitro studies showed EPO secretion from modified HPMC over 6 months. Intraperitoneal transplantation aided with collagen matrix was the most effective. EPO, in MPMC transplanted mice, was detected up to 3 weeks (peak at 13 +/- 1 mIU/mL), and anemia of uremic mice was corrected (35.3 +/ 0.9 mIU/mL to 41.9 +/- 1.1 mIU/mL). CONCLUSION: PF-HPMC can be considered as an appropriate target for gene therapy since these cells can be efficiently isolated, modified, and transplanted. Nevertheless, implantation techniques in the peritoneum should be directed at obtaining longer duration of transgene expression in vivo, and means should be developed for enabling regulated expression of the gene. PMID- 12753298 TI - Glomerular number and size in autopsy kidneys: the relationship to birth weight. AB - BACKGROUND: In the Southeast United States, African Americans have an estimated incidence of hypertension and end-stage renal disease (ESRD) that is five times greater than Caucasians. Higher rates of low birth weight (LBW) among African Americans is suggested to predispose African Americans to the higher risk, possibly by reducing the number of glomeruli that develop in the kidney. This study investigates the relationships between age, race, gender, total glomerular number (Nglom), mean glomerular volume (Vglom), body surface area (BSA), and birth weight. METHODS: Stereologic estimates of Nglom and Vglom were obtained using the physical disector/fractionator combination for autopsy kidneys from 37 African Americans and 19 Caucasians. RESULTS: Nglom was normally distributed and ranged from 227,327 to 1,825,380, an 8.0-fold difference. A direct linear relationship was observed between Nglom and birth weight (r = 0.423, P = 0.0012) with a regression coefficient that predicted an increase of 257,426 glomeruli per kilogram increase in birth weight (alpha = 0.050:0.908). Among adults there was a 4.9-fold range in Vglom, and in adults, Vglom was strongly and inversely correlated with Nglom (r =-0.640, P = 0.000002). Adult Vglom showed no significant correlation with BSA for males (r = -0.0150, P = 0.936), although it did for females (r = 0.606, P = 0.022). No racial differences in average Nglom or Vglom were observed. CONCLUSION: Birth weight is a strong determinant of Nglom and thereby of glomerular size in the postnatal kidney. The findings support the hypothesis that LBW by impairing nephron development is a risk factor for hypertension and ESRD in adulthood. PMID- 12753299 TI - The AGE inhibitor pyridoxamine inhibits lipemia and development of renal and vascular disease in Zucker obese rats. AB - BACKGROUND: In previous studies, pyridoxamine (PM) limited the formation of advanced glycation end products (AGEs) and development of nephropathy in streptozotocin-diabetic rats without affecting glycemic control. However, the lipid-lowering effects of PM and the correlation of plasma cholesterol and triglycerides with AGEs in skin collagen suggested that lipids might be an important source of AGEs in the diabetic rat. This study addresses the effects of hyperlipidemia on formation of advanced glycation and lipoxidation end products (AGE/ALEs) and the effects of PM on hyperlipidemia, hypertension, AGE/ALE formation, and development of nephropathy in the nondiabetic, Zucker obese rat. METHODS: Three groups of Zucker rats were studied: lean (Fa/fa), untreated fatty (fa/fa), and fa/fa treated with PM (2 g/L drinking water). Blood pressure, plasma lipids and creatinine, and urinary albumin were measured monthly. AGE/ALEs were measured in skin collagen by high-performance liquid chromatography (HPLC) and gas chromatography/mass spectrometry (GC/MS). Changes in wall thickness of the aorta and renal arterioles were evaluated by light microscopy. RESULTS: AGE/ALEs formation was increased two- to threefold in skin collagen of obese versus lean rats. PM inhibited the increases in AGE/ALEs in collagen, and significantly decreased the rise in plasma triglycerides, cholesterol, and creatinine, corrected hypertension and thickening of the vascular wall, and nearly normalized urinary protein and albumin excretion in Zucker fa/fa rats. CONCLUSION: Lipids are an important source of chemical modification of tissue proteins, even in the absence of hyperglycemia. PM inhibited AGE/ALE formation and hyperlipidemia and protected against renal and vascular pathology in a nondiabetic model. PMID- 12753300 TI - Cell senescence in rat kidneys in vivo increases with growth and age despite lack of telomere shortening. AB - BACKGROUND: Somatic cells in vitro have a finite life expectancy before entering a state of senescence, but it is unclear whether this state occurs in vivo in kidney development, growth, and aging. We previously showed that human kidney cortex displays telomere shortening with age. In this study, we compared the structural and functional changes in rat kidney with age to phenomena associated with cellular senescence in vitro. METHODS: We assessed the changes in Fischer 344 rat kidneys from age 1 to 9 months to define growth and development and from age 9 to 24 months to define aging. RESULTS: Rat kidney telomeres were approximately 35 to 40 kb long and did not shorten significantly. Expression of mRNA for p16INK4a, a characteristic senescence gene in vitro, was undetectable in most young rats but rose 27 fold during growth and a further 72-fold during aging. p16INK4a protein was localized to the nucleus and increased with age. p16INK4a mRNA also increased in other tissues. Lipofuscin and senescence associated beta-galactosidase increased in epithelium with growth and aging and their occurrence was significantly associated with each other. Lipofuscin was particularly found in atrophic nephrons. CONCLUSION: We conclude that cell senescence occurs in both growth and aging in rat kidney and may contribute to the age-related pathology. These changes are not due to telomere shortening, but may reflect cumulative environmental stress. PMID- 12753301 TI - Shortening of telomeres: Evidence for replicative senescence of T cells derived from patients with Wegener's granulomatosis. AB - BACKGROUND: Replicative senescence describes the fact that somatic cells undergo a finite and predictable number of cell divisions before entering an irreversible state of growth arrest. Progressive shortening of the telomeres, a consequence of cell division, is a reliable indicator of replicative senescence. METHOD: We analyzed telomere length of DNA derived from T cells of patients suffering from Wegener's granulomatosis by Southern blotting. Moreover, expression of CD28, another marker for replicative senescence, was tested by cytofluorometry. RESULTS: In patients with disease for more than 5 years, short telomeres were detected in addition to telomeres of normal length, indicating replicative senescence of discrete T-cell clones. Reduced expression of CD28 was noted, particularly on CD8-positive T cells, derived from patients with disease for more than 5 years and short telomeres. CONCLUSION: Our data provide evidence that a portion of T cells had undergone replicative senescence, which in turn indicates clonal expansion of T cells as consequence of activation. PMID- 12753302 TI - Reduced tubular cation transport in diabetes: prevented by ACE inhibition. AB - BACKGROUND: The renal clearance of organic cations is important for the homeostasis of a number of exogenous and endogenous compounds. The organic cation transporters (OCTs) situated on the basolateral surface of proximal tubular cells mediate active cation excretion. Alterations of cation transport may occur in diabetes, although the role of the OCTs has not been previously assessed. METHODS: Experimental diabetes was induced in rats with streptozotocin (55 mg/kg) and animals were randomly assigned to receive ramipril (3 mg/mL) in drinking water for 24 weeks. In a second protocol, rats were infused with angiotensin II (Ang II) at a dose of 58.3 ng/kg/min for 2 weeks via an implanted osmotic pump. Expression of the OCTs and renal clearance of the endogenous cation N-methyl nicotinamide (NMN) was assessed. RESULTS: Diabetes was associated with a reduction in gene and protein expression of both OCT-1 and OCT-2 and a reduction in NMN clearance. These effects were prevented by ramipril, associated with the prevention of albuminuria and tubular injury as demonstrated by the expression of osteopontin and glutathione peroxidase 3 (GPX-3). An infusion of Ang II also reduced NMN clearance but without altering the renal expression of OCTs. CONCLUSION: We hypothesize that reduced expression of OCTs in diabetes may be a marker of tubular injury. However, Ang II may also directly augment renal cation clearance independent of changes in transporter expression. Together these effects may provide additional mechanism to explain treatment-related improvements in creatinine clearance and renoprotection in diabetes following blockade of the renin-angiotensin system (RAS). PMID- 12753303 TI - Hypertension superimposed on type II diabetes in Goto Kakizaki rats induces progressive nephropathy. AB - BACKGROUND: Type II diabetes in the Goto Kakizaki (GK) rats (derived from Wistar rats) is not associated with the development of obesity, hyperlipidemia, hypertension, or pronounced renal functional changes. The aim of this study was to investigate the effect of superimposed hypertension on renal function and morphology under conditions of hyper- and normoglycemia. METHODS: The evolution of biochemical and morphologic renal changes was examined in GK and Wistar rats treated with deoxycorticosterone acetate (DOCA) salt over 24 weeks. RESULTS: Blood pressure was increased from 6 weeks on in GK and Wistar rats with no difference in blood pressure levels between both groups (week 24, 183 +/- 14 mm Hg vs. 191 +/- 13 mm Hg, P = NS, vs. 144 +/- 6 mm Hg in normal controls, P < 0.01). A progressive increase in proteinuria was observed in hypertensive GK rats from 12 weeks on (week 24, 168 +/- 62 mg/day vs. 41 +/- 30 mg/day in hypertensive Wistar rats, P = 0.002). Histologic analysis at weeks 15 and 24 showed progressive glomerulosclerosis in hypertensive GK and Wistar rats (week 24, 13 +/ 4% vs. 8 +/- 1%, P = NS) but not in nonhypertensive GK controls. This was associated with evidence of podocyte damage (de novo desmin expression) in hypertensive as compared to nonhypertensive GK rats (week 24, score 1.4 +/- 0.1 vs. 0.8 +/- 0.1, P < 0.001) while no significant increase was observed in hypertensive vs. nonhypertensive Wistar rats. Tubulointerstitial damage was increased in hypertensive GK as compared to hypertensive Wistar rats (week 24, score 1.5 +/- 0.6 vs. 0.6 +/- 0.3, P = 0.01). By immunohistochemistry, this was associated with an up-regulation of tubulointerstitial type IV collagen as well as alpha-smooth muscle actin (alpha-SMA) expression, macrophage infiltration and cell proliferation in hypertensive GK rats. CONCLUSION: Our data demonstrate that long-standing type II diabetes alone is not sufficient to induce progressive nephropathy unless secondary injurious mechanisms such as hypertension are present. The hypertensive GK rat provides a novel model to investigate the mechanisms involved in diabetic nephropathy. PMID- 12753304 TI - Very low-density lipoprotein-apoprotein CI is increased in diabetic nephropathy: comparison with apoprotein CIII. AB - BACKGROUND: Recent studies have suggested that apoprotein (apo) CI in very low density lipoprotein (VLDL) plays an important role in causing hypertriglyceridemia independent of apo CIII, which is associated with coronary heart disease (CHD). Because the incidence of CHD is increased in diabetic patients and is even higher when diabetic nephropathy is developed, we measured apo CI levels in VLDL from type 2 diabetic patients, with various degree of nephropathy, and compared the results with those for healthy controls or nondiabetic patients with chronic renal failure (CRF). METHODS: This study enrolled healthy control subjects, type 2 diabetic patients with normoalbuminuria, microalbuminuria, overt proteinuria, and CRF on hemodialysis and nondiabetic hemodialysis patients. VLDL (density <1.006) was separated by ultracentrifugation. Then the apo CI, CIII, and B concentrations in VLDL were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: The apo CI, CIII, and B concentrations in VLDL were respectively 3-, 2-, and 2-fold higher, respectively, in diabetic patients with overt proteinuria than in controls. Hemodialysis patients with diabetic nephropathy had levels of apo CI, CIII, and B in VLDL that were 2.6-, 2.7- and 2-fold higher, respectively, than those in controls. Nondiabetic hemodialysis patients also had a 2.7-fold higher level of VLDL apo CIII, whereas VLDL apo CI and VLDL apo B were not significantly increased. VLDL apo CI was significantly correlated with VLDL apo B independently of VLDL apo CIII level. CONCLUSION: An increase of VLDL apo CIII is a prominent feature of dyslipidemia in CRF patients, regardless of whether they are diabetic or nondiabetic, whereas an increase of VLDL apo CI is more specific to diabetic nephropathy and is closely associated with an increase of VLDL particle numbers, a new risk factor for CHD. PMID- 12753305 TI - Variations in cell signaling pathways for different vasoconstrictor agonists in renal circulation of the rat. AB - BACKGROUND: Major cell signaling pathways involved in agonist-induced vasoconstriction are recognized to be Ca2+ mobilization via inositol-1,4,5 triphosphate (IP3), Ca2+ influx through l-type channels, activation of protein kinase C (PKC), and of Rho-associated kinase (ROK). However, their contribution for renal vasoconstriction induced by different agonists is not well characterized. METHODS: Increasing doses of angiotensin II (Ang II), norepinephrine, and arginine vasopressin (AVP) were infused into the left renal artery of anesthetized rats to reduce renal blood flow from a threshold value to about 50%. Rightward shift of the dose-response curves due to coinfusion of inhibitors served to assess contribution of different pathways: trimethoxybenzoate (TMB-8) against Ca2+ mobilization, nifedipine against Ca2+ influx, staurosporine and Ro-318220 against PKC, and Y-27632 and HA-1077 against ROK. Effects of inhibitors were also determined for renal response to a single dose of U-46619, a thromboxane A2 agonist. Composite response to U-46619 consisting of a fast and slow component did not permit determination of dose response curves. RESULTS: Inhibition of ROK by Y-27632 or HA-1077 had the largest effect on renal responses to agonists. They shifted dose-response curves of Ang II, norepinephrine, and AVP to sevenfold and higher values. Staurosporine, nifedipine, and TMB-8 had variable effect on agonist responses. They attenuated effects of Ang II and norepinephrine in an additive manner, and each of them increased effective dose values about fourfold. TMB-8 did not attenuate response to AVP and U-46619. Staurosporine and nifedipine diminished effects of AVP in a nonadditive manner, and attenuated additively the fast component of U-46619 response. CONCLUSION: In contrast to other cell signaling pathways, ROK plays a common role for all vasoconstrictor agonistsis in renal circulation. PMID- 12753306 TI - Contributions of nitric oxide, EDHF, and EETs to endothelium-dependent relaxation in renal afferent arterioles. AB - BACKGROUND: Acetylcholine-induced endothelium-dependent relaxation in the renal afferent arteriole has been ascribed to nitric oxide, but the role of endothelium derived hyperpolarizing factors (EDHFs) and 14,15-epoxyeicosatrienoic acid (14,15 EET) are unclear. METHODS: Single afferent arterioles were dissected from kidney of normal rabbits and microperfused in vitro at 60 mm Hg. Vessels were preconstricted submaximally with norepinephrine (10(-8) mol/L). Relaxation was assessed following cumulative addition of ACh (10(-9) to 10(-4) mol/L) alone, or in the presence of indomethacin (to inhibit cyclooxygenase), Nw-nitro-L-arginine (L-NNA) (to inhibit nitric oxide synthase), methylene blue (to inhibit soluble guanylate cyclase), or a combination of L-NNA + methylene blue. To assess contributions by EDHF, studies were repeated with either apamin + charybdotoxin [to block Ca2+-activated K+ channels (KCa)] or with 40 mmol/L KCl. To asses the role of 14,15-EET, relaxations were evaluated in the presence of its competitive inhibitor 14,15-epoxyeicosa-5(Z)-enoic acid (14,15-EEZE). RESULTS: Relaxation by acetylcholine was abolished following endothelial denudation. It was unaffected by indomethacin but was inhibited 54%+/- 5% (P < 0.001) by L-NNA, 57%+/- 5% by methylene blue, and 60%+/- 4% by the combination of L-NNA plus methylene blue. Relaxation was inhibited further by KCl (80%+/- 6%) or by apamin + charybdotoxin (96%+/- 2%). 14,15-EEZE, alone, inhibited acetylcholine-induced relaxation by 29%+/- 3%, and by 80%+/- 5% in the presence of L-NNA. CONCLUSION: Acetylcholine induced afferent arteriolar relaxation depends strongly on both nitric oxide, acting via soluble guanylate cyclase, and on an EDHF, likely 14,15-EET, acting via KCa. PMID- 12753307 TI - Hypocitraturia is one of the major risk factors for nephrocalcinosis in very low birth weight (VLBW) infants. AB - BACKGROUND: Very low birth weight (VLBW) infants are at risk to develop nephrocalcinosis (NC). NC may result from spontaneous or therapy-induced imbalance between promoters and inhibitors of crystallization in the urine. However, data on "normal" urinary excretions of these parameters in VLBW infants are sparse. Therefore, we prospectively examined the urinary excretion of calcium, oxalate, uric acid, and citrate in VLBW infants during the first 8 weeks of life. METHODS: Urine samples were collected once weekly in 124 VLBW infants. NC appeared in 16 infants, whose data were separately analyzed. The remaining 108 infants were divided into subgroups: A, <1000 g (N = 53); and B, 1000 to 1500 g (N = 55). Random urine samples were analyzed and the results were expressed as molar creatinine ratios. Calcium/citrate and oxalate/citrate expressed the risk for calcium oxalate crystallization. RESULTS: In group A, citrate excretion was lower at weeks 2 to 5 and 7; calcium/citrate was higher in weeks 2, 4, and 7; oxalate/citrate was higher in weeks 3, 4, 7, and 8; and calcium/creatinine ratio was higher in week 4 (P < 0.05). Citrate/creatinine ratios were low in nine infants with NC. Oxalate/creatinine and calcium/creatinine were elevated in five and calcium/citrate was increased in nine infants with NC. CONCLUSION: Hypocitraturia is a major risk factor for NC in VLBW infants, especially in those <1000 g. The urinary excretions in VLBW infants seem to depend on birth weight, age, and clinical condition. Hence, supplementation with alkali citrate may have a beneficial effect in the prevention of NC. PMID- 12753308 TI - Cross-sectional and prospective data on urinary calcium and urinary stone disease. AB - BACKGROUND: Urinary calcium is considered a risk factor for urinary stone disease (USD), although prospective data are missing. This epidemiologic study investigates cross-sectionally and longitudinally the relation of urinary calcium excretion to USD. METHODS: In the Gubbio Population Study, data on USD were collected by questionnaire during medical examinations from 1989 to 1992 (baseline) and telephone interviews in 1997 to 1998 (follow-up). Baseline data collection included overnight urinary calcium excretion and use of medications. Study cohort was made of 1458 men and 1799 women, age 25 to 74 years, and not on treatment with diuretics at baseline. USD was diagnosed by: excretion of stone(s), and/or radiographic or ultrasonic evidence, and/or surgical or endoscopic removal of stone(s). RESULTS: At baseline, urinary calcium excretion was higher in persons with than without USD (215 and 182 micromol/hour, P < 0.001) and related to USD prevalence independent of gender, age, and other variables (P < 0.001). Among persons without USD at baseline, baseline urinary calcium excretion was higher in persons with than without incident USD at follow up (202 and 181 micromol/hour, P = 0.034) and related to incident USD independent of gender, age, and other variables. A difference of 100 micromol/hour (about 1 SD) in urinary calcium excretion related to a difference in USD risk of 1.32 for prevalence and 1.21 for incidence (95% CI = 1.15/1.52 and 1.01/1.45, respectively) in multivariate analyses controlled for gender, age, body mass index, parental history of USD, urinary excretion of urea, sodium, and potassium. CONCLUSION: Cross-sectional and prospective data show that urinary calcium excretion is a risk factor for USD. PMID- 12753309 TI - Peroxynitrite-induced oxidation of plasma lipids is enhanced in stable hemodialysis patients. AB - BACKGROUND: The relationship between end-stage renal disease (ESRD), hemodialysis, and oxidative stress is controversial. To determine whether ESRD causes oxidative stress, we measured basal levels of plasma F2-isoprostanes as a marker of lipid peroxidation in vivo, and peroxynitrite-stimulated formation of F2-isoprostanes, as a marker of the oxidizibility of plasma lipids in vitro, before and after routine hemodialysis. METHODS: Total plasma F2-isoprostanes were measured by gas chromatography-mass spectrometry (GC-MS) before and after the oxidation of plasma lipids with the peroxynitrite-generating compound, 3 morpholino-sydnonimine (SIN-1), in 23 patients with ESRD patients undergoing regular hemodialysis, and 14 controls. Plasma vitamin E concentrations were measured by high-performance liquid chromatography (HPLC). RESULTS: There was no difference in basal plasma concentrations of F2-isoprostanes in the ESRD group prior to hemodialysis, 246 +/- 20 pg/mL, compared to controls, 252 +/- 28 pg/mL, or immediately on completion of hemodialysis, 236 +/- 14 pg/mL. Incubation of control plasma with SIN-1 caused the formation of F2-isoprostanes with plasma concentrations increasing to 987 +/- 54 pg/mL at 6 hours. The formation of F2 isoprostanes stimulated by SIN-1 was markedly enhanced in the plasma obtained from patients undergoing hemodialysis at 1861 +/- 174 pg/mL, P < 0.001, and SIN-1 induced formation of F2-isoprostanes was further increased in plasma obtained immediately after hemodialysis at 2437 +/- 168 pg/mL, P < 0.001. Incubation of plasma with SIN-1 resulted in the net consumption of vitamin E. CONCLUSION: Although basal plasma F2-isoprostanes were similar in patients with ESRD compared with controls, the presence of oxidative stress in patients with ESRD was unmasked when the plasma was stressed by peroxynitrite generated from SIN-1, and this was enhanced further by hemodialysis. PMID- 12753310 TI - Maximal suppression of renin-angiotensin system in nonproliferative glomerulonephritis. AB - BACKGROUND: Elimination of residual proteinuria is the novel target in renoprotection; nevertheless, whether a greater suppression of renin-angiotensin system (RAS) effectively improves the antiproteinuric response in patients with moderate proteinuria remains ill-defined. METHODS: We evaluated the effects of maximizing RAS suppression on quantitative and qualitative proteinuria in ten patients with stable nonnephrotic proteinuria (2.55 +/- 0.94 g/24 hours) due to primary nonproliferative glomerulonephritis (NPGN), and normal values of creatinine clearance (103 +/- 17 mL/min). The study was divided in three consecutive phases: (1) four subsequent 1-month periods of ramipril at the dose of 2.5, 5.0, 10, and 20 mg/day; (2) 2 months of ramipril 20 mg/day + irbesartan 300 mg/day; and (3) 2 months of irbesartan 300 mg/day alone. RESULTS: Maximizing RAS suppression was not coupled with any major effect on renal function and blood pressure; conversely, a significant decrement in hemoglobin levels, of 0.8 g/dL on average, was observed during up-titration of ramipril dose. The 2.5 mg dose of ramipril significantly decreased proteinuria by 29%. Similar changes were detected after irbesartan alone (-28%). The antiproteinuric effect was not improved either by the higher ramipril doses (-30% after the 20 mg dose) or after combined treatment (-33%). The reduction of proteinuria led to amelioration of the markers of tubular damage, as testified by the significant decrement of alpha 1 microglobulin (alpha 1m) excretion and of the tubular component of proteinuria at sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). CONCLUSION: In nonnephrotic NPGN patients, standard doses of either ramipril or irbesartan lead to significant reduction of residual proteinuria and amelioration of the qualitative features suggestive of tubular damage. The enhancement of RAS suppression up to the maximal degree does not improve the antiproteinuric response and is coupled with a decrement of hemoglobin levels. PMID- 12753311 TI - Patterns of hepatitis B prevalence and seroconversion in hemodialysis units from three continents: the DOPPS. AB - BACKGROUND: Hepatitis B (HBV) historically has been a public health issue within hemodialysis units. This study estimates HBV prevalence and seroconversion rates across seven countries and investigates associations with facility level practice patterns. METHODS: The study sample was from the Dialysis Outcomes and Practice Patterns Study (DOPPS), a cross-sectional, prospective, observational study of adult hemodialysis patients randomly selected from 308 dialysis facilities in France, Germany, Italy, Spain, the United Kingdom, Japan, and the United States. Logistic regression was used to model the odds ratio (OR) of HBV prevalence, and Cox regression was used to model time from entry into the study to HBV seroconversion. RESULTS: In this sample, mean HBV facility prevalence was 3.0% with a median of 1.9%. The percentage of facilities with an HBV prevalence 0% to 5% was 78.5%. Adjusted HBV prevalence was higher in France, Germany, and Italy and lower in Japan and the United Kingdom. The majority of facilities (78.1%) had a seroconversion rate of 0 conversions per 100 patient-years. Presence of a protocol for HBV-infected patients was significantly associated with HBV seroconversion in the separate practice pattern model [risk ratio (RR) = 0.52, P = 0.03] and in the combined practice pattern model (RR = 0.44, P = 0.01). CONCLUSION: There are differences in HBV prevalence and rate of seroconversion both at the country and the hemodialysis facility level. Presence of a protocol for HBV-infected patients was strongly and significantly associated with decreased risk for seroconversion. The observed variation suggests opportunities for improved HBV outcomes with further definition of optimal practice patterns at the facility level. PMID- 12753312 TI - Microsatellite DNA polymorphism of human adrenomedullin gene in type 2 diabetic patients with renal failure. AB - BACKGROUND: Adrenomedullin (AM) is a hypotensive peptide widely produced in the cardiovascular organs and tissues such as the heart, kidney, and the vascular cells. We have previously cloned and sequenced the genomic DNA encoding human AM gene, and determined that the gene is located in the short arm of chromosome 11. The 3'-end of the gene is flanked by the microsatellite marker of cytosine adenine (CA) repeats. In this study, we investigated the association between DNA variations in AM gene and the predisposition to develop nephropathy in type 2 diabetes mellitus. METHODS: Genomic DNA was obtained from the peripheral leukocytes of 233 normal healthy subjects (NH), 139 type 2 diabetic patients on hemodialysis (DM-HD), 106 control patients with type 2 diabetes without nephropathy (DM-C) and 318 hemodialysis patients due to chronic glomerulonephritis (CGN-HD). The genomic DNA was subject to polymerase chain reaction (PCR) using a fluorescence-labeled primer, and the number of CA repeats were determined by polyacrylamide gel electrophoresis (PAGE). RESULTS: In our Japanese subjects, there existed four types of alleles with different CA-repeat number; 11, 13, 14, and 19. The frequencies of these alleles were 11: 27.7%, 13: 32.8%, 14: 35.6%, and 19: 3.9% in NH. These allele frequencies were not significantly different in DM-C and CGN-HD. However, DM-HD showed significantly different distribution of allele frequency from other groups (chi 2 = 18.9, P = 0.026). Namely, the frequency of 19-repeat allele in DM-HD was higher (9.0%) than NH, DM-C, and CGN-HD (P = 0.005, 0.041, and 0.004, respectively). CONCLUSION: The microsatellite DNA polymorphism of AM gene may be associated with the genetic predisposition to develop nephropathy in Japanese patients with type 2 diabetes mellitus. PMID- 12753313 TI - Hepatitis C virus-related cryoglobulinemic glomerulonephritis: long-term remission after antiviral therapy. AB - BACKGROUND: Renal involvement in patients with hepatitis C virus (HCV) infection commonly manifests as cryoglobulinemic glomerulonephritis (CGN). The combination of interferon-alpha (IFN-alpha) and ribavirin, which is currently considered the standard antiviral therapy in chronic hepatitis C, could be difficult to carry out in cryoglobulinemic patients who are frequently anemic, even in the absence of renal failure. Clinical and histologic long-term results of this therapeutic regimen have not been so far reported in patients with CGN. METHODS: Three patients with HCV-related CGN and slightly impaired kidney function were treated with IFN-alpha and ribavirin for 12 months, and subsequently were followed up for 24 to 36 months. Two of these patients who were anemic were pretreated with erythropoietin (EPO). In each patient renal biopsy was performed before starting therapy and repeated 14 to 26 months after the end of treatment. RESULTS: In all three patients, antiviral therapy induced sustained virologic response, which was followed by clear improvement in clinical, biochemical, immunologic, and histologic features. Clinical and biochemical improvement steadily progressed in all three patients, achieving normal or nearly normal results at the end of follow-up. In contrast, some immunologic features, such as serum levels of C4 and rheumatoid factor activity, did not normalize in two and three patients, respectively. Posttreatment renal biopsies showed mildly active histologic lesions. CONCLUSION: Antiviral therapy with IFN-alpha and ribavirin may be considerably beneficial in patients with HCV-related CGN who obtain sustained virologic response. PMID- 12753314 TI - A novel HIV-1 transgenic rat model of childhood HIV-1-associated nephropathy. AB - BACKGROUND: A characteristic finding of human immunodeficiency virus (HIV) associated nephropathy (HIVAN) is the presence of heavy proteinuria, focal or global glomerulosclerosis, and microcystic tubular dilatation leading to renal enlargement, and rapid progression to end-stage renal disease (ESRD). METHODS: We have recently developed the first HIV-1 transgenic rat model that carry a noninfectious HIV-1 DNA construct lacking 3.1 kb of sequence overlapping the gag and pol sequences, and develop many of the clinical lesions seen in HIV-infected patients, including HIVAN. To gain further insight into the pathogenesis of childhood HIVAN, we followed the clinical and renal pathologic outcome of 165 HIV 1 transgenic (HIV-Tg) rats and their respective control littermates for a period of 18 months. RESULTS: HIV-1 Tg rats progressively developed proteinuria and renal histologic lesions similar to those seen in children with HIVAN, leading to chronic renal failure. By in situ hybridization, HIV-1 genes were detected in glomerular and tubular epithelial cells and infiltrating mononuclear cells, which also expressed the HIV-1 envelop protein gp120. The development of HIVAN was associated with the accumulation of basic fibroblast growth factor (bFGF) in the kidney. CONCLUSION: These data support the notion that HIV-1 plays a direct role in the pathogenesis of HIVAN, by affecting the function and growth of renal epithelial cells, inducing the recruitment of mononuclear cells, and accumulating bFGF in the kidney, even in the absence of viral replication. These rats may provide an excellent model system to study the pathogenesis of childhood HIVAN. PMID- 12753315 TI - Retarding progression of chronic renal disease: the neglected issue of residual proteinuria. AB - BACKGROUND: Findings that early changes in proteinuria independently predict long term glomular filtration rate (GFR) decline (Delta GFR) would highlight proteinuria as a major determinant of progression in chronic renal disease. METHODS: We investigated whether percent changes (3 months vs. baseline) in proteinuria (adjusted for concomitant changes in GFR) and residual proteinuria at 3 months, predicted Delta GFR [over a median (IQ range) follow up of 31.3 (24.5 to 50.3) months] in 273 patients with proteinuric chronic nephropathies enrolled in the Ramipril Efficacy In Nephropathy (REIN) study. RESULTS: Short-term changes and residual proteinuria (r = -0.23, P = 0.0001 for both) significantly correlated with Delta GFR and, at multivariate analyses, independently predicted Delta GFR (beta = -0.23, P = 0.0002; beta = -0.21, P = 0.0004, respectively). For comparable levels of residual proteinuria, patients with greater short-term reduction had slower Delta GFR (-0.28 +/- 0.04 mL/min/1.73 m2/ vs. -0.53 +/- 0.07 mL/min/1.73 m2/month, P = 0.04). On ramipril and conventional treatment, specular short-term changes in proteinuria (-18.2 +/- 3.5% vs. 24.2 +/- 6.7%, P < 0.0001, respectively) were associated with significantly different Delta GFRs. However, similar changes in proteinuria resulted in a difference in Delta GFR (ramipril, 0.39 +/- 0.07 mL/min/1.73 m2/month; conventional therapy, 0.74 +/- 0.11 mL/min/1.73 m2/month; P < 0.01) that was sevenfold higher (0.35 vs. 0.05 mL/min/1.73 m2/month) in patients with basal proteinuria > or =3 g/24 hours as compared to those with basal proteinuria 1 to 3 g/24 hours (ramipril, 0.25 +/- 0.06 mL/min/1.73 m2/month; conventional therapy, 0.30 +/- 0.07 mL/min/1.73 m2/month; P = NS). CONCLUSION: Regardless of blood pressure control and treatment randomization, short-term changes in proteinuria and residual proteinuria reliably predict long-term disease progression. Reducing proteinuria is renoprotective, particularly in nephrotic patients. As for arterial hypertension, proteinuria should be a specific target for renoprotective treatment. PMID- 12753316 TI - Elevation of blood thioredoxin in hemodialysis patients with hepatitis C virus infection. AB - BACKGROUND: Thioredoxin (TRX) is a stress-inducible thiol-containing protein, which has been shown to be an indicator of oxidative stress in a variety of diseases. The association between oxidative stress and hepatitis C virus (HCV) infection, however, remains unknown in hemodialysis patients. METHODS: We measured serum TRX levels in 85 hemodialysis patients positive for anti-HCV antibodies (age, 60 +/- 1 years old; hemodialysis duration, 17 +/- 1 years; M/F = 57/28) by enzyme-linked immunosorbent assay (ELISA), and examined whether blood TRX may be associated with HCV-related hepatic injury. RESULTS: Serum TRX was significantly higher in hemodialysis patients with HCV infection (112.3 +/- 3.7 ng/mL, N = 85) than in those without HCV infection (69.7 +/- 3.3 ng/mL, N = 59) (age, 69 +/- 2 years old; hemodialysis duration, 6 +/- 1 years; M/F = 32/27, P < 0.01) or normal subjects (28.0 +/- 5.4 ng/mL, N = 9). TRX was significantly correlated with time on hemodialysis (r = 0.27, P = 0.01) in HCV-positive patients, while it was associated with the patient's age in HCV-negative patients (r = 0.42, P < 0.01). Blood TRX was significantly correlated with asparate aminotransferase in patients with HCV infection (r = 0.34, P < 0.01) and without HCV infection (r = 0.46, P < 0.01). However, serum TRX was not associated with blood alanine aminotransferase, a relatively specific marker of hepatic cellular damage, in HCV-infected hemodialysis patients. A significant relationship was found between serum ferritin and TRX (r = 0.25, P = 0.02) and malondialdehyde (MDA) values (r = 0.25, P = 002) in HCV-positive patients. Serum TRX was also higher in the patients receiving weekly iron supplement with HCV infection (135.3 +/- 10.2 ng/mL vs. 110.2 +/- 3.9 ng/mL, P = 0.06) and without HCV infection (91.8 +/- 12.1 ng/mL vs. 65.2 +/- 2.7 ng/mL, P < 0.01). CONCLUSION: There was a greater increase in serum TRX in hemodialysis patients with HCV viremia than without HCV viremia. However, there may not be an association between serum TRX and HCV related hepatic injury. TRX increased with serum ferritin in HCV-infected patients and further increased by iron infusion. These findings indicate that HCV infection and iron loading may aggravate oxidative stress in dialysis patients. PMID- 12753317 TI - Effects of raloxifene on bone metabolism and serum lipids in postmenopausal women on chronic hemodialysis. AB - BACKGROUND: Premature amenorrhea and hypoestrogenism and lack of hormone replacement therapy after menopause have been frequently reported in uremic women on dialysis. Therefore, in addition to renal osteodystrophy, postmenopausal women on dialysis could be at risk of osteoporosis. In addition, these patients are at higher risk for hyperlipidemia, arteriosclerosis, and subsequent coronary heart disease and stroke. Recent evidence has suggested that hormone replacement therapy (HRT) in postmenopausal women could have several beneficial effects as well as potentially serious risks. Great efforts have been made to identify therapeutic alternatives that would have the benefits of estrogen on brain and bone without its adverse effects on breast and endometrium. In the present study, we evaluated the effect of raloxifene, a selective estrogen receptor modulator (SERM), on bone metabolism and serum lipids in postmenopausal women on chronic hemodialysis. METHODS: We performed a prospective, blind, placebo-controlled, and randomized study. Fifty postmenopausal women on chronic hemodialysis with proven severe osteopenia or osteoporosis by bone densitometry were selected. After a written informed consent, patients were randomized into two groups: 25 women on placebo and 25 women on the study drug, raloxifene hydrochloride, at a dose of 60 mg/day. In all patients, we performed a baseline bone mineral density (BMD) analysis and simultaneously evaluated different biochemical parameters, serum lipids (total low-density lipoprotein [LDL] and high-density lipoprotein [HDL] cholesterol and triglycerides) and serum markers of bone resorption (pyridinoline crosslinks). BMD was reassessed after 1 year of therapy. Bone resorption markers were determined every 3 months for 1 year. RESULTS: After 1 year on raloxifene therapy, lumbar spine BMD (trabecular bone) significantly improved, whereas femoral neck BMD (cortical bone) did not change significantly. No changes in BMD were observed at trabecular or cortical sites in the placebo group. Serum pyridinoline levels showed a significant decrease after 6 months on raloxifene that persisted thereafter. Low-density lipoprotein (LDL)-cholesterol decreased significantly in the raloxifene group with no changes in serum triglycerides, total cholesterol, or HDL cholesterol. No significant side effects were observed in the raloxifene group. CONCLUSION: The study demonstrates that after one year on raloxifene, postmenopausal women on hemodialysis have a significant increase in trabecular BMD, decrease in bone resorption markers and LDL-cholesterol values, suggesting that SERMs could constitute a therapeutic alternative to improve bone metabolism and control of hyperlipidemia in these patients. The possible long-term effects of raloxifene remain to be determined. PMID- 12753318 TI - Long-term effects on bone mineral density of pamidronate given at the time of renal transplantation. AB - BACKGROUND: Fracture rate after renal transplantation is substantially increased, is a source of morbidity and mortality, and correlates with osteopenia. The rate of bone loss after transplantation is time dependent. While we recorded marked bone loss during the first year after renal transplantation, bone loss in long term recipients (>24 months) was found to be similar to expected age-related decline. We have previously shown that treatment with pamidronate at the time of transplantation protected the skeleton over a 1-year study period. METHODS: We have reexamined patients who participated in our original study, all of whom had been randomized to receive either placebo or pamidronate (0.5 mg/kg) at the time of transplantation and 1 month later. We now report 4-year data from 17 of the 26 original cohort. All patients received immunosuppression, comprising prednisolone, cyclosporine, and azathioprine. RESULTS: We found that without prophylaxis bone loss at 4 years was substantial and significant at the femoral neck (mean loss was -12.3%) but was not significant at the lumbar spine (mean loss was -4.64%). Patients who received two doses of pamidronate experienced no statistically significant bone loss at either the femoral neck or the lumbar spine. Patient characteristics of the placebo and treatment groups were similar with the exception of serum parathyroid hormone concentrations, which remained higher at 4 years in the pamidronate-treated patients (15.8 +/- 3.7 pmol/L vs. 9.8 +/- 1.8 pmol/L, P < 0.05). CONCLUSION: Without prophylaxis, most patients who continue to receive low dose glucocorticoids as part of maintenance immunosuppression manifest a substantial deficit in bone mineral density (BMD) at the femoral neck. In contrast, two doses of pamidronate given at the time of transplantation and 1 month later protected the skeleton from significant bone loss over the 4 years after transplantation. PMID- 12753319 TI - Effects of the glutamate carboxypeptidase II (GCP2 1561C>T) and reduced folate carrier (RFC1 80G>A) allelic variants on folate and total homocysteine levels in kidney transplant patients. AB - BACKGROUND: The effect of the glutamate carboxypeptidase II GCP2 1561C>T and the reduced folate carrier 1 RFC1 80G>A polymorphisms on folate and total homocysteine (tHcy) plasma levels of kidney transplant patients are unknown. METHODS: In a cross-sectional study of 730 kidney allograft recipients, GCP2 1561C>T, RFC1 80G>A, folate, and tHcy plasma levels were analyzed using linear regression models that allowed dependent covariates to follow a gamma distribution for univariate and multivariate analyses. RESULTS: The allele frequency for GCP2 1561C>T was 0.05, and 0.43 for RFC1 80G>A. Heterozygosity or homozygosity for GCP2 1561C>T was associated with higher folate plasma levels compared to patients without mutation (P < 0.0001), while RFC1 80G>A showed no influence. Multiple testing, also including MTHFR 677C>T and MTHFR 1298A>C, revealed no interaction between the different genotypes and the folate plasma concentration. Neither GCP2 1561C>T nor RFC1 80G>A showed an association with tHcy plasma levels. CONCLUSION: We conclude that GCP2 1561C>T is associated with elevated folate levels. GCP2 1561C>T and RFC1 80G>A are no major determinants of tHcy plasma levels in kidney transplant patients. PMID- 12753320 TI - Incidence of latent mesangial IgA deposition in renal allograft donors in Japan. AB - BACKGROUND: Mesangial immunoglobulin A (IgA) deposition is incidentally encountered in asymptomatic individuals, but its precise frequency and significance had not been clarified. The background of the latent IgA deposition is related to the epidemiology and pathogenesis of IgA nephropathy. METHODS: Zero hour allograft biopsies were performed in 510 renal transplantations (446 living donors, and 64 cadaveric donors) at the Kidney Center of Tokyo Women's Medical University. Mesangial IgA and C3 deposition were analyzed immunohistochemically, and the frequency and clinicopathologic features of mesangial IgA deposition were investigated. RESULTS: Mesangial IgA deposition was present in 82 (16.1%) of the total 510 allografts with no statistical difference between living donors (72/446, 16.1%) and cadaveric donors (10/64, 15.6%) or between blood-related donors (66/392, 16.8%) and nonblood-related donors (16/110, 14.5%). Mesangial C3 deposition was present in 16 (19.5%) of the 82 allografts with mesangial IgA deposition. The grade of hematuria in IgA(+) donors was significantly higher than IgA(-) donors (1.30 +/- 1.17 vs. 0.86 +/- 0.89, P = 0.025). Histologic investigation of IgA(+) allografts revealed the frequency of mesangioproliferative glomerulonephritis (PGN) was significantly higher in IgA(+)/C3(+) allografts (8/16, 50%) than in IgA(+)/C3(-) allografts (11/66, 16.7%) (P = 0.0084). Moreover, the number of infiltrated macrophages to glomerulus (cells/glomerular cross section) was significantly higher in the IgA(+)/C3(+) allografts than in IgA(+)/C3(-), IgA(-)/C3(+) and IgA(-)/C3(-) allografts (1.10 +/- 0.62 vs. 0.61 +/- 0.42, P = 0.0008; 0.47 +/- 0.34, P = 0.023; and 0.37 +/- 0.23, P = 0.002, respectively). CONCLUSION: The latent mesangial IgA deposition was a relatively common phenomenon in the healthy Japanese donors. This phenomenon was associated with mild degree of microhematuria, mesangial proliferation and glomerular macrophage infiltration in some of the affected individuals, especially with combined IgA and C3 deposition. PMID- 12753321 TI - The clinical characteristics and antiretroviral dosing patterns of HIV-infected patients receiving dialysis. AB - BACKGROUND: Human immunodeficiency virus (HIV)-related renal disease is the third leading cause of end-stage renal disease (ESRD) among African Americans aged 24 to 60 years. This study describes the clinical characteristics and antiretroviral dosing patterns of HIV-infected patients receiving dialysis to define the clinical needs of this growing population. METHODS: Demographic and clinical information was collected on all HIV-infected patients incident to dialysis after January 1, 1998 until January 1, 2001 at five medical centers. The cohort was described overall and by subgroups based on hepatitis status, CD4 lymphocyte count, and use of antiretroviral therapy. Continuous and categoric variables were compared using either the Wilcoxon rank sum or Student t test and Fisher's exact or chi-square tests, as appropriate. RESULTS: A total of 89 patients were included, 55 of whom were alive at the time of data collection. The mean age was 44.6 years (range, 22.7 to 66.9 years), 74.2% were male, and 83.2% patients were African Americans. While only 45.9% of patients undergoing renal biopsy were diagnosed with HIV-associated nephropathy (HIVAN), the majority of patients who had not undergone biopsy carried the clinical diagnosis of HIVAN (69.8%, P = 0.03). Of the cohort, 19.7% tested hepatitis B surface antigen positive, and 67.1% had reactive antibody tests for hepatitis C. Patients with hepatitis C were more likely to have experienced intravenous drug use as a risk behavior for HIV acquisition (OR 8.2; 95% CI 2.39, 27.9; P = 0.001] and to be older (OR 1.1 per year of age; 95% CI 1.02, 1.2; P = 0.01). A total of 60.7% of patients were receiving antiretroviral medication at last follow-up. Among patients alive and receiving antiretroviral medications at the time of data collection, absolute CD4+ count rose (268 vs. 339 cells/mL, P = 0.03), while among patients alive, but not receiving antiretroviral medications, absolute CD4+ count did not change (389 vs. 392 cells/mL, P = 0.11) during similar periods of follow-up. No difference was seen between initial and current HIV RNA levels for either group. Among patients receiving antiretroviral medications, there were significant variations in dosing regimens. The greatest variation was seen in the prescribing patterns of lamivudine with a 12-fold difference among patients. CONCLUSION: The projected growth of the HIV-infected ESRD population requires a better understanding of the clinical needs of this population. The high prevalence of coinfection with hepatitis C as well as the wide variations in dosing patterns for antiretroviral medications are areas that require further investigation to minimize morbidity and mortality among this group. PMID- 12753322 TI - Hypertension increases expression of growth factors and MHC II in chronic allograft nephropathy. AB - BACKGROUND: Hypertension of the recipient is strongly associated with chronic allograft nephropathy. It is unclear, however, whether hypertension is the cause or the consequence of chronic allograft nephropathy. METHODS: The present study was performed in the Fisher to Lewis rat kidney transplant model. Transplanted rats (N = eight in each group) received either no treatment or were made hypertensive by administration of deoxycorticosteron acetate (DOCA) and salt. Proteinuria and systolic blood pressure was measured monthly, grafts were harvested at 3 and 6 months for semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) and for immunohistology. RESULTS: Systolic blood pressure was markedly elevated in rats receiving DOCA/salt. Allografts of hypertensive animals contained significantly more cells expressing the proliferating cell nuclear antigen compared to isografts and to allografts from normotensive animals (P < 0.05). Histologic staining and mRNA expression of major histocompatibility complex (MHC) II was markedly increased in allografts of hypertensive animals compared to all other groups (P < 0.05). Expression of mRNA for platelet-derived growth factor-B (PDGF-B), transforming growth factor-beta (TGF-beta) and collagen was higher in allografts than in isografts and was highest in hypertensive animals. CONCLUSION: We conclude that hypertension augments the expression of growth factors in the allograft possibly aggravating the intimal hyperplasia observed in chronic allograft nephropathy. By increasing the expression of MHC II antigens, hypertension may render the allograft more susceptible to alloantigen dependent damage. Hypertension and alloantigen-dependent factors appear to exert additive or synergistic effects on inflammatory pathways leading to graft injury. PMID- 12753324 TI - A new partnership between TGF-beta1 and glucocorticoids in the network of inflammation. PMID- 12753323 TI - Health-related fitness and quality of life following steroid withdrawal in renal transplant recipients. AB - BACKGROUND: Exercise capacity increases significantly soon after transplantation; however, over time it does not further improve and patients remain low compared to normal levels. The limitations to exercise following transplantation have not been identified, but may be related to immunosuppression therapy regimens that include prednisone. METHODS: We studied health-related fitness measures (cardiorespiratory fitness, muscle strength, and body composition) and quality of life in renal transplant recipients randomized into two groups: those using standard maintenance immunosuppression, including prednisone therapy (N = 14); and those undergoing rapid withdrawal of steroids using Simulect[interleukin-2 (IL-2) receptor inhibitor] (N = 9). Testing was done at 3 and 12 months following transplant and the 12-month data were compared to 15 normal sedentary controls. RESULTS: Compared to those maintained on steroids, the steroid withdrawal group showed greater gains in VO2peak (P = 0.05) and quadriceps peak torque (P = 0.05) and greater gains in the vitality score and the Physical Composite Scale on the SF-36 questionnaire (P < 0.05). At 1 year, all patients had significantly lower exercise capacity compared to the sedentary controls (P = 0.01). No differences were observed in body composition, with both patient groups increasing in body weight (primarily body fat) over time. At 12 months, all patients were not different in body fat percentage compared to the sedentary controls. CONCLUSION: We conclude that prednisone is not the cause for increased body fat following transplantation; however, it may contribute to lower spontaneous improvements in exercise capacity possibly by limiting increases in muscle strength. The low exercise capacity in all transplant recipients studied at 1 year suggests a need for exercise training to optimize physical functioning following transplant. PMID- 12753325 TI - Clinical and cellular markers of diabetic nephropathy. PMID- 12753326 TI - Presentation of the 2001 A.N. Richards Award to Eberhard Fromter. PMID- 12753327 TI - Acceptance of the 2001 A.N. Richards Award. PMID- 12753328 TI - Presentation of the 2001 Jean Hamburger Award to Professor Anita Aperia. PMID- 12753329 TI - Acceptance of the 2001 Jean Hamburger Award. PMID- 12753332 TI - Who should receive hepatitis A vaccine? AB - Though a potent vaccine represents a powerful preventive tool, the policy of its use is governed by epidemiological and economical factors. Hepatitis A, an enterically transmitted disease shows distinct association with socio-economic status, populations with improvement experiencing lower exposure to the virus. With the availability of vaccine, it is pertinent to consider its use in the effective control of the disease. However, with the varied epidemiological patterns and economical constraints in different countries it does not seem to be possible to evolve universal policy for immunization. Though, universal immunization may be the most effective way of control, the same is not practical for many countries. It is proposed that irrespective of endemicity of hepatitis A, high-risk groups such as travelers to endemic areas, patients suffering from chronic liver diseases, HBV and HCV carriers, tribal communities with high HBV carrier rates, food handlers, sewage workers, recipients of blood products, troops, and children from day-care centers should be immunized with hepatitis A vaccine. In addition, for populations with intermediate prevalence, infants, children from affordable families may be immunized. As coupling the vaccine with EPI schedule would be beneficial, use of combined A & B or A, B & E vaccine may be an attractive alternative. PMID- 12753333 TI - The role of intrahepatic immune effector cells in inflammatory liver injury and viral control during chronic hepatitis B infection. AB - Cytotoxic T lymphocytes (CTL) and Kupffer cells play an important role in the immune control of hepatitis B virus (HBV), but may also induce liver injury during infection. We investigated the intrahepatic immune response in liver biopsies of chronic HBV patients in relation to inflammatory liver injury and viral control. Forty-seven liver biopsies from patients with chronic HBV with varying degrees of inflammation (ALT values) were selected. Acute hepatitis and normal liver specimens served as controls. Immune effector cells, cytotoxic effector molecules and cytokine producing cells were quantified after immunohistochemical staining in lobular and portal areas of the biopsies. The intralobular number of CD8+ T-lymphocytes was significantly decreased in biopsies of patients with high ALT (r = -0.54; P < 0.001). Higher ALT-values were correlated with increased numbers of granzyme+ cells in portal areas (r = 0.65; P < 0.001) and higher numbers of intralobular Fas-L+ cells (r = 0.32; P = 0.05). Fas-L was expressed on Kupffer and lymphoid cells. More intralobular CD8+ T lymphocytes were found in HBeAg- than in HBeAg+ patients (P = 0.002). But IFN gamma and TNF-alpha producing cells were observed sporadically in chronic HBV patients. Hence, in chronic HBV infection, low viral replication and HBeAg negativity is related to increased presence of intralobular CD8+ T-lymphocytes. Persistence of the virus may be caused by the absence of cells producing anti viral cytokines in the liver. Inflammatory liver injury during chronic HBV infection is probably not the result of increased numbers of infiltrating CD8+ T lymphocytes, but of Fas-L expression by Kupffer cells and increased cytolytic activity of cells in portal areas. PMID- 12753334 TI - Increasing serum levels of IgM anti-HCV are diagnostic of recurrent hepatitis C in liver transplant patients with ALT flares. AB - Recurrent hepatitis and acute rejection share common features which make difficult for diagnosis in liver transplant hepatitis C virus (HCV) positive patients. We studied the usefulness of quantitative monitoring of HCV RNA and immunoglobulin (Ig)M anti-HCV in the differential diagnosis between recurrent hepatitis and acute rejection in 98 consecutive anti-HCV positive liver transplant patients. Aminotransferase levels, serum HCV RNA and IgM anti-HCV were measured at the time of transplantation and monthly thereafter. A liver biopsy (LB) was obtained when serum aminotransferase levels increased to twice or more than normal. During a mean follow-up of 16 months 86 aminotransferase flares were observed. Histology was compatible with recurrent hepatitis C in 44 cases and with acute rejection in 28, doubtful in 14. The fluctuations of HCV RNA serum levels were not significantly different in the three groups. Serum IgM anti-HCV levels increased (from negative to positive or with value variations > or = 0.18) in 36 of 44 cases with recurrent hepatitis C at the time of alanine aminotransferase (ALT) flare. IgM anti-HCV remained unchanged in all rejection cases (P < 0.001), but increased in 10 of 11 histologically doubtful cases that were diagnosed as hepatitis at the second LB. Increasing serum levels of IgM anti HCV at the time of ALT flares are significantly associated with recurrent hepatitis C in liver transplant patients. The quantitative monitoring of IgM anti HCV appears to be an additional diagnostic tool for distinguishing recurrent hepatitis C from acute graft rejection with a 100% specificity; 100% positive predictive value and 88.9% diagnostic accuracy. PMID- 12753335 TI - Chronic hepatitis C patients with a post-treatment virological relapse re-treated with an induction dose of 18 MU interferon-alpha in combination with ribavirin and amantadine: a two-arm randomized pilot study. AB - Thirty-seven chronic hepatitis C patients with virological relapse (VR) after previous interferon-alpha (IFN) or IFN/ribavirin (Riba) therapy, were re-treated. Patients were randomized for either IFN/Riba and amantadine (Ama) including a 2 week initial high IFN induction course (18 MU IFN daily) (group A) or the same 2 week IFN induction course combined with Riba/Ama, followed by Riba/Ama without IFN (group B). Treatment duration for both groups was 24 weeks with a 24-week follow-up thereafter. The inclusion in group B was prematurely stopped because all patients (n = 10) relapsed within 2 weeks after stopping IFN. Therefore, all subsequent patients were included in group A (n = 27). In group A, 44% achieved a sustained virological response (SVR) and 29% of the patients with an end-of treatment virological response had a VR again. Of all pretreatment characteristics, only genotype non-1 patients had a significantly higher chance of achieving SVR (P < 0.001). Of the characteristics during treatment only a negative hepatitis C virus (HCV)-RNA test result in transcription-mediated amplification (TMA) at week 6 had a high predictive value for SVR, 80% in all patients and 92% in genotype non-1 patients. In conclusion, hepatitis C patients with a VR to previous antiviral treatment can be successfully re-treated with IFN induction combined with Riba/Ama for only 6 months, when they have genotype non-1 and a negative HCV-RNA test result in TMA 6 weeks after the start of therapy. Riba/Ama combination therapy without IFN does not prevent VR after 2 weeks high IFN induction. PMID- 12753336 TI - Influence of age and date of infection on distribution of hepatitis C virus genotypes and fibrosis stage. AB - The objective of this study was to assess the influence of age and date of acquisition of hepatitis C virus (HCV) infection on the distribution of genotypes and the progression of fibrosis in HCV-infected patients who were born in Spain and had their habitual place of residence in this country. Genotypic analysis was performed in 375 patients in whom it was possible to establish the year of HCV infection because the mode of transmission was known (transfusion, injection drug use, blood donor, or epidemic outbreak). In 298 patients with liver biopsy, fibrosis stage was related to age at infection, duration of infection, alcohol consumption, and HCV genotype. HCV subtype 1b was almost exclusively detected among transfusion recipients, but the onset of intravenous drug addiction was associated with the introduction of HCV genotypes other than 1b among injecting users with subsequent spread to other exposure risk groups. Fibrosis progression was influenced by alcohol consumption, increased duration of infection, and older age at infection. In summary, spread of intravenous drug use determined HCV infection by genotypes other than 1b. The risk of fibrosis progression was influenced more by age at viral acquisition and alcohol consumption than by the infecting genotype. PMID- 12753337 TI - A randomized trial to assess the efficacy of interferon alpha in combination with ribavirin in the treatment of interferon alpha nonresponders with chronic hepatitis C: superior efficacy of high daily dosage of interferon alpha in genotype 1. AB - A randomized trial was conducted to assess the efficacy of daily (QD) or thrice weekly (TIW) administration of interferon-alpha (IFN) in high doses in combination with ribavirin (1.0-1.2 g/day) in patients with chronic hepatitis C (CHC) who were nonresponders to previous IFN monotherapy. Interferon was administered as 10 MU IFN (QD or TIW) for 4 weeks, followed by 5 MU IFN (QD or TIW) for 20 weeks, and then by 3 MU IFN (QD or TIW) for 24 weeks. Sustained virological response (SVR) was evaluated in 142 patients who received at least one dose of medication. One-fourth of the patients achieved SVR, 26% of those treated with IFN QD and 25% of those treated with IFN TIW (P = 0.85). For genotype 1 patients, SVR rates were 32.4 and 15.8% for IFN QD and IFN TIW, respectively, whereas for genotype non-1 patients the corresponding SVR rates were 20.6 and 36.4%, respectively (test of homogeneity: P = 0.031). This finding was further confirmed by multivariate logistic regression analysis where a statistically significant interaction (P = 0.012) was found between treatment and HCV genotype indicating that the IFN QD regimen was superior to IFN TIW among genotype 1 patients whereas, among genotype non-1 patients, the two treatments were similar (odds ratio of SVR in IFN QD vs IFN TIW: 3.33 among genotype 1 patients, 95% CI: 1.00-11.14). In conclusion, re-treatment of patients not responding to previous IFN monotherapy with a combination of high daily dose of IFN with ribavirin may be beneficial for genotype 1 infected patients. PMID- 12753338 TI - Reinforced interferon alpha-2b and ribavirin is more effective than standard combination therapy in the retreatment of chronic hepatitis C previously nonresponsive to interferon: a randomized trial. AB - Interferon-alpha (IFN) monotherapy results in sustained virological clearance in a minority of patients with chronic hepatitis C. The aim of this study was to assess the effect of a reinforced regimen combining ribavirin and high-dose IFN for 48 weeks compared with a nonreinforced regimen combining a standard IFN regimen and ribavirin for 24 weeks in nonresponders with chronic hepatitis C. A total of 231 patients with chronic hepatitis C and previous nonresponse to IFN monotherapy were randomized. The reinforced group (n = 114) received IFN-2b 6 million units (MU) thrice weekly (TIW) and ribavirin for 48 weeks, and the nonreinforced group (n = 117) received IFN-2b 3 MU TIW and ribavirin for 24 weeks. The main outcome measure was a sustained virological response, defined as negative serum hepatitis C virus (HCV)-RNA 24 weeks following the end of treatment. This endpoint was determined in 98 patients of the reinforced group and 105 patients of the nonreinforced group. At the end of follow-up, a sustained virological response was observed in 29 of the 98 patients (29.6%) in the reinforced group vs 16 of the 105 patients (15.2%) in the nonreinforced group (P = 0.014). In multivariate analysis, factors associated with a sustained virological response were treated with a reinforced regimen [odds ratio (OR) 2.9; P = 0.06] and genotype 2 or 3 (OR 8.8; P < 0.0002). A total of 160 patients had paired biopsies before and after treatment. Histological activity improvement was observed in 32 of 80 patients (40%) and fibrosis worsening in 26 of 80 patients (33%) in the reinforced group vs 13 of 80 (16%) and 19 of 80 (24%) in the nonreinforced group (P = 0.30 and 0.20, respectively). Hence in nonresponders, a high-dose 48-week regimen of IFN and ribavirin combination was more effective than a regimen with interferon at lower dose and ribavirin for 24 weeks only. PMID- 12753339 TI - High sustained virological response in chronic hepatitis C by combining induction and prolonged maintenance therapy. AB - Chronic hepatitis C patients with genotype 1 infection, liver cirrhosis, high viral load, or those who have not responded to anti-viral treatment in the past have limited chances of clearing the virus, even with pegylated interferon ribavirin therapy. In this study we treated such patients with a treatment schedule that combines high dose induction Interferon (IFN), prolonged daily IFN and ribavirin treatment. Twenty-four consecutive patients were included in this study with either genotype 1 infection, cirrhosis, previous non-response to IFN or a combination of these poor-response characteristics. Patients were treated with 10 million units (MU) of IFN daily for 4 weeks followed by 5 MU/day until week 24, 3 MU/day until week 52 and 3 MU thrice weekly until week 76 in combination with 1-1.2 g ribavirin daily. HCV RNA levels were assessed weekly until week 4 and at least once every 3 months thereafter, by a validated assay with a detection limit below 500 copies/mL. Both intention to treat (ITT) and per protocol (PP) analysis showed a high sustained virological response (ITT 67%, PP 80%). A virological response occurred rapidly (before 8 weeks of treatment) in all patients with a sustained response. Relapse after stopping therapy was observed in only 5%. Side-effects were observed frequently, and six patients had to be hospitalized. With this new treatment regimen that combines induction- and prolonged daily interferon treatment with ribavirin it seems possible to eliminate hepatitis C virus in the majority of patients that have an a priori limited chance of sustained response. Further clinical evaluation of intensive interferon and ribavirin combination therapy (now also including PEG-interferon) is recommended in centres that can provide close patient monitoring and experienced hepatological support. PMID- 12753340 TI - Interferon alpha in the treatment of chronic hepatitis C in children: a meta analysis [correction of metanalysis]. AB - Children with chronic hepatitis C may be ideal candidates for treatment with interferon alpha (IFNalpha) as they have liver disease at an early stage. However, adverse drug reactions need to be considered. The aim of this study was to conduct a systematic review of literature on interferon therapy of chronic hepatitis C in children, and to perform a meta-analysis of pooled data. A computerized search gave 18 articles on IFNalpha therapy in children with chronic hepatitis C; after exclusion of uncontrolled trials and of trials including patients with comorbidities, data from two studies could be pooled (48 patients). The outcomes assessed were biochemical, defined as normalization of alanine transaminase, and virologic, defined as HCV-RNA loss, both sustained at 24 months after enrollment. Results of the studies were homogenous. Risk difference was 37% (95%CI: 12.9-61) in favour of IFNalpha treated children for sustained biochemical response, and 36.8% (95%CI: 14.3-59.3) in favour of treated children for sustained HCV clearance, respectively. The differences were highly significant (P = 0.007 and P = 0.004, respectively). The histological end-point, as well as side effects, could not be analysed, due to lack of data. This review identifies the poor methodology of the majority of the published trials. The study provides support for the efficacy of IFNalpha in improving both biochemical and virologic outcomes in chronic hepatitis C in children, but evidence is confined to these surrogate end-points. PMID- 12753342 TI - Aetiology and prognostic factors in acute liver failure in India. AB - The early prognostic indicators for acute liver failure in endemic zones for hepatitis E virus have not been determined. All consecutive patients with acute liver failure from a geographically defined region endemic for hepatitis E virus were studied over the period April 1989-April 1996. Demographic, clinical and biochemical parameters were recorded at presentation and serum samples were analysed for known viral hepatitis (A-E) markers. Multiple parameters were compared in survivors and non-survivors in a univariate analysis. All significant factors on univariate analysis were entered into a stepwise logistic regression analysis to identify independent variables of prognosis. The sensitivity and specificity of significant prognostic factors was then assessed. A total of 180 [69 males and 111 females: age (mean +/- SD) 31.1 +/- 14.7 years] with acute liver failure were studied. Of these, 131 (72.8%) patients died. Hepatitis E virus was the aetiological cause in 79 (43.9%) patients, while hepatitis A virus, hepatitis B virus, hepatitis C virus and non-A, non-E agent/'s could be incriminated in four (2.1%), 25 (13.9%), 13 (7.2%) and 56 (31.1%) patients respectively. Of 83 women in childbearing age, 49 (59.0%) were pregnant, 33 (67.3%) of these were in the third trimester. Forty-seven (95.8%) pregnant women had HEV infection. Nine variables differed significantly between survivors and non-survivors on univariate analysis. Of these, four variables which predicted the adverse outcome on multivariate analysis were non-hepatitis-E aetiology, prothrombin time >30 s, grade of coma >2 and age >40 years in that order of significance. Pregnancy per se or duration of gestation did not adversely affect the prognosis. In endemic areas, hepatitis E virus is the commonest cause of acute liver failure. Acute liver failure occurs in a high proportion of pregnant women, mostly in third trimester. Early predictors of a poor outcome are non-E aetiology, prothrombin time >30 s, grade of coma >2 and age >40 years. PMID- 12753343 TI - Alarming prevalence of hepatitis-B infection among the Jarawas--a primitive Negrito tribe of Andaman and Nicobar Islands, India. AB - Jarawas, a classical hunter-gatherer tribe of Andaman and Nicobar islands have lived in isolation for several centuries. It is only recently have they started to come in contact with the outsiders, shedding their hostility. Since then, several disease outbreaks have been reported amongst them. Screening of sera samples collected during one such outbreak showed very high endemicity of hepatitis-B infection with over 60% of the individuals positive for hepatitis B surface antigen (HBsAg). The rates of HBsAg observed among the Jarawas are probably the highest ever reported in the world and warrant immediate control measures which would prevent further spread of this infection in the community. PMID- 12753341 TI - Retreatment with interferon and ribavirin vs interferon alone according to viraemia in interferon responder-relapser hepatitis C patients: a prospective multicentre randomized controlled study. AB - Low pretreatment viral load has consistently been shown to be an independent predictor of sustained response (SR) in patients with chronic hepatitis C infection. We assessed the efficacy of interferon (IFN) plus ribavirin vs IFN alone in low viraemic patients (<2 millions copies/mL) who had relapsed to a previous course of IFN and the efficacy of 24 vs 48 week combination therapy in high viraemic patients. Two hundred and ninety-seven patients were randomly assigned to one of the four regimens after stratification on pretreatment viral load. All patients received IFN-alpha2b (6 million units thrice weekly for 24 weeks and 3 million units thrice weekly for 24 weeks). Patients with low viraemia received either IFN-alpha2b alone for 48 weeks (R1: 42 patients) or IFN-alpha2b plus ribavirin (600 mg/day) for 24 weeks and IFN-alpha2b alone for the next 24 weeks (R2: 48 patients). Patients with high viral load received either IFN alpha2b plus ribavirin for 24 weeks and then IFN-alpha2b alone for the next 24 weeks (R3: 104 patients) or IFN-alpha2b plus ribavirin for 48 weeks (R4: 103 patients). In low viraemic patients the rate of SR was 37.7% in group R1 and 59.6% in group R2 (P < 0.05). In high viraemic patients, the rate of SR was 44.7% in group R3 and 51.4% in group R4 (P: NS). Thirty-one patients discontinued treatment (10.4%) without difference regarding treatment regimen. In the regimen using ribavirin we found no difference in terms of SR between patients receiving a dose of ribavirin below 10.6 mg/kg/day (55%) or over 10.6 mg/kg/day (58%). Histological improvement occurred in 70.2% of patients regardless of the regimen. Logistic regression showed that genotype 2 and 3, Knodell score <6 and alanine aminotransferase pretreatment level >3 x upper limit of normal were significantly and independently correlated with SR. In low viraemic patients who relapsed to a previous IFN treatment, combination therapy using high-dose IFN and low-dose ribavirin is better than high-dose IFN alone. In high viraemic patients there was no benefit in increasing the duration of combination therapy from 24 to 48 weeks. In this study, it was found that low dose of ribavirin can be used safely and there is no effect of ribavirin dose on SR. PMID- 12753344 TI - Evidence of patient-to-patient transmission of hepatitis C virus through contaminated intravenous anaesthetic ampoules. AB - Two separate cases of acute hepatitis C virus (HCV) infection following medical procedures, arthroscopy and colonoscopy, are reported. In both episodes, patient risk factors were reviewed, and staff and other patients' sera were tested for HCV antibodies and RNA. HCV RNA positive samples were genotyped, sequenced, and subjected to phylogenetic analysis. No risk factors for HCV infection were identified for either case except for medical procedures. HCV RNA positive patients were identified preceding both cases on the respective theatre lists. HCV infection in a second low risk patient was also identified. Nucleic acid sequencing and phylogenetic analysis of HCV from the two putative source patients and the three recipient patients demonstrated a high degree of relatedness respectively. The results suggest that patient-to-patient transmission occurred in both episodes via contamination of intravenous anaesthetic ampoules with HCV used on multiple patients. Injectable medication ampoules should not be used for more than one patient. PMID- 12753345 TI - Actigraphy suggests age-related differences in napping and nocturnal sleep. AB - The aim of this study was to contrast the time distribution of out-of-bed napping in young and older adults through recordings of wrist activity, and to evaluate the correlates of napping with nocturnal sleep. Seventy-three young adults between 18 and 32 years and 60 older adults between 60 and 75 years of age participated in the study. Subjects were selected for good general health and had few sleep complaints. They wore wrist-activity monitors and kept daily sleep logs for 1 week. Automatic sleep scoring was edited by the authors, supplemented by sleep logs and illumination data as well as activity data. Napping episodes were modestly increased in older adults, but there was no difference in the daily duration of napping. Older adults napped more in the evening (especially within 2 h before bedtime), whereas young adults napped more in the afternoon. The older adults with evening naps (n = 31) showed earlier nocturnal wake-up times and decreased nocturnal sleep duration compared with the older adults without evening naps (n = 29). There was no difference in nocturnal sleep between young adults with afternoon naps (n = 32) and without afternoon naps (n = 41). In determining the effects of napping on nocturnal sleep, timing of napping and age are important. Maintaining alertness during the evening (e.g. by bright light exposure or moderate exercise) would be a possible approach to delay wake-up times in older adults. PMID- 12753346 TI - Daytime sleepiness during Ramadan intermittent fasting: polysomnographic and quantitative waking EEG study. AB - During the lunar month of Ramadan, Muslims abstain from eating, drinking and smoking from sunrise to sunset. We reported previously that Ramadan provokes a shortening in nocturnal total sleep time by 40 min, an increase in sleep latency, and a decrease in slow-wave sleep (SWS) and rapid eye movement (REM) sleep duration during Ramadan. During the same study, the effects of Ramadan intermittent fasting on daytime sleepiness were also investigated in eight healthy young male subjects using a quantitative waking electroencephalograph (EEG) analysis following the multiple sleep latency test (MSLT) procedure. This procedure was combined with subjective alertness and mood ratings and was conducted during four successive experimental sessions: (1) baseline (BL) 15 days before Ramadan, (2) beginning of Ramadan (R11) on the 11th day of Ramadan, (3) end of Ramadan (R25) on the 25th day of Ramadan, (4) recovery 2 weeks after Ramadan (AR). During each session, four 20-min nap opportunities (MSLTs) were given at 10:00, 12:00, 14:00 and 16:00 h and were preceded by rectal temperature readings. Nocturnal sleep was recorded before each daytime session. Subjective daytime alertness did not change in R25 but decreased in R11 at 12:00 h, and subjective mood decreased at 16:00 h, both in R11 and R25. During the MSLT, mean sleep latency decreased by an average of 2 min in R11 (especially at 10:00 and 16:00 h) and 6 min in R25 (especially at 10:00 and 12:00 h) compared with BL. There was an increase in the daily mean of waking EEG absolute power in the theta (5.5-8.5 Hz) frequency band. Significant correlations were found between sleep latency during the MSLT and the waking EEG absolute power of the fast alpha (10.5 12.5 Hz), sigma (11.5-15.5 Hz) and beta (12.5-30 Hz) frequency bands. Sleep latency was also related to rectal temperature. In conclusion, Ramadan diurnal fasting induced an increase in subjective and objective daytime sleepiness associated with changes in diurnal rectal temperature. PMID- 12753347 TI - Sleep-wake rhythm in an irregular shift system. AB - Sleep in shift work has been studied extensively in regular shift systems but to a lesser degree in irregular shifts. Our main aim was to examine the sleep-wake rhythm in shift combinations ending with the night or the morning shift in two irregular shift systems. Three weeks' sleep/work shift diary data, collected from 126 randomly selected train drivers and 104 traffic controllers, were used in statistical analyses including a linear mixed model and a generalized linear model for repeated measurements. The results showed that the sleep-wake rhythm was significantly affected by the shift combinations. The main sleep period before the first night shift shortened by about 2 h when the morning shift immediately preceded the night shift as compared with the combination containing at least 36 h of free time before the night shift (reference combination). The main sleep period before the night shift was most curtailed between two night shifts, on average by 2.9 and 3.5 h among the drivers and the controllers, respectively, as compared with the reference combination. Afternoon napping increased when the morning or the day shift immediately preceded the night shift, the odds being 4.35-4.84 in comparison with the reference combination. The main sleep period before the morning shift became 0.5 h shorter when the evening shift preceded the morning shift in comparison with the sleep period after a free day. The risk for dozing off during the shift was associated only with the shift length, increasing by 17 and 35% for each working hour in the morning and the night shift, respectively. The results demonstrate advantageous and disadvantageous shift combinations in relation to sleep and make it possible to improve the ergonomy of irregular shift systems. PMID- 12753348 TI - The effect of sleep loss on next day effort. AB - The study had two primary objectives. The first was to determine whether sleep loss results in a preference for tasks demanding minimal effort. The second was to evaluate the quality of performance when participants, under conditions of sleep loss, have control over task demands. In experiment 1, using a repeated measures design, 50 undergraduate college students were evaluated, following one night of no sleep loss and one night of sleep loss. The Math Effort Task (MET) presented addition problems via computer. Participants were able to select additions at one of five levels of difficulty. Less-demanding problems were selected and more additions were solved correctly when the participants were subject to sleep loss. In experiment 2, 58 undergraduate college students were randomly assigned to a no sleep deprivation or a sleep deprivation condition. Sleep-deprived participants selected less-demanding problems on the MET. Percentage correct on the MET was equivalent for both the non-sleep-deprived and sleep-deprived groups. On a task selection question, the sleep-deprived participants also selected significantly less-demanding non-academic tasks. Increased sleepiness, fatigue, and reaction time were associated with the selection of less difficult tasks. Both groups of participants reported equivalent effort expenditures; sleep-deprived participants did not perceive a reduction in effort. These studies demonstrate that sleep loss results in the choice of low-effort behavior that helps maintain accurate responding. PMID- 12753349 TI - Cardiac autonomic characteristics in infants sleeping with their head covered by bedclothes. AB - The risk of Sudden Infant Death Syndrome is increased in infants sleeping with their head covered by bedding items. This study was designed to evaluate cardiac autonomic nervous controls in infants sleeping with the head covered by bedclothes. Sixteen healthy infants with a median age of 12 weeks (range 9-13 weeks) were recorded polygraphically for one night. While they slept in their usual supine position, a bedsheet was placed over their head for about 45 min. All infants were challenged with the head covered and with the head free during both rapid eye movement (REM) and non-rapid eye movement (NREM) sleep. Sleep, breathing and heart rate (HR) characteristics were recorded simultaneously, together with rectal and pericephalic temperatures. In both head-free and head covered conditions, autoregressive spectral analysis of HR was evaluated as a function of sleep stages. During the head-covered periods, parasympathetic tonus decreased and sympathetic activity increased in both REM and NREM sleep. Compared with the head-free periods, the head-covered sleep periods were characterized by greater rectal (P = 0.012) and pericephalic temperatures (P = 0.002). Covering the infant's head with a bedsheet was associated with significant changes in autonomic balance. The finding could be related to an elevation in temperatures within the infant's microenvironment. PMID- 12753350 TI - Factors of home dream recall: a structural equation model. AB - Previous research has indicated that personality factors such as openness to experience, creativity, visual memory, attitude toward dreams, and sleep behavior is related to home dream recall frequency (DRF). However, a study investigating all areas simultaneously within one sample in order to determine the percentage of variance explained by all variables and to take intercorrelations between the influencing factors into account has not been performed till now. The present study with 444 participants fills this gap. Using several indicators for each of the variables mentioned above, a structural equation model was tested. Although the model fit was satisfying, the four factors which were significantly related to DRF: personality (openness to experience, thin boundaries, absorption), creativity, nocturnal awakenings, and attitude toward dreams, explained only 8.4% of the total variance. As this value is considerably lower than those of studies investigating a single influencing factor and using similar measurement instruments in similar samples, one might speculate about possible expectancy effects in these previous studies, an effect which has been demonstrated for DRF in the laboratory setting. In addition, the small percentage of explained variance of each single factors (<3%) may indicate that other, in this study unmeasured, variables such as sleep duration (state aspect), introspection, and cognitive functioning immediately upon awakening (sleep inertia) show substantial covariance with the interindividual differences in DRF. Future studies should focus on longitudinal aspects in order to differentiate between state versus trait factors (although methodologic issues, e.g. the effect of the measurement technique on DRF itself, have to be clarified) and investigate additional variables which might be associated with DRF (see above). PMID- 12753351 TI - Sensitivity of sleep stages to painful thermal stimuli. AB - Many modalities of both acute and chronic pain have been shown to disrupt sleep. Any differences in the intensity of thermal noxious stimulus required to produce arousal from stage 2, slow-wave sleep (SWS) and rapid eye movement (REM) sleep is unclear. An assessment of reactions of seven male (age 22 +/- 2.9 years) and three female subjects (age 21.0 +/- 1.0 years) to a range of gradually increasing temperatures was used both when awake and asleep. When awake, subjects assigned five different descriptors to the increasing heat stimulus. During the different stages of sleep, temperatures were increased over the same range as when awake until the subjects aroused from sleep. The possible fluctuations in pain perception due to a time-of-night effect were assessed in awake subjects over a 12-h period from 19:00 to 07:00 hours. During sleep, arousals occurred at significantly higher temperatures during SWS (P < 0.01) and REM sleep (P < 0.05) than during stage 2 sleep. The temperatures causing arousals during SWS and REM sleep were not significantly different and were equivalent to temperatures causing pain tolerance when awake. No changes in pain perception due to time of night were observed. The results show that a higher intensity of thermal noxious stimulus is required to cause arousal from SWS and REM sleep when compared with stage 2 sleep. This would confirm the suspicion that REM sleep and SWS are relatively, and possibly equally, resistant to disruption by noxious stimuli. PMID- 12753352 TI - The locus coeruleus complex of the bottlenose dolphin (Tursiops truncatus) as revealed by tyrosine hydroxylase immunohistochemistry. AB - Using tyrosine hydroxylase immunohistochemistry we examined the structure of the pontine, or rostral rhombencephalic, catecholaminergic cells groups, which may be collectively termed the locus coeruleus complex (LC), in the bottlenose dolphin. The present study is the first to describe the LC in a cetacean species and, at 1.3 kg, represents the largest non-human brain to date in which the LC has been investigated. We identified four catecholaminergic cell groups in the dorsal pontine tegementum and peri-aqueductal gray matter: A6 dorsal (locus coeruleus), A6 ventral (locus coeruleus alpha), A7 (subcoeruleus), and A5 (fifth arcuate nucleus). No patterns of cellular distribution, nuclear subdivision, or cellular morphology indicate specialization of the LC, which might have been anticipated because of the large absolute brain size and unihemispheric sleep phenomenology of cetaceans. PMID- 12753353 TI - Practicality of the Sleepstrip in postal screening for obstructive sleep apnoea. AB - The SleepStrip, a disposable screening device for the detection of obstructive sleep apnoea (OSA), which displays an estimated apnoea-hypopnoea index (AHISS) was posted, with instructions, to 48 patients referred for sleep study for suspected OSA. The patients subsequently underwent a cardiorespiratory sleep study from which the AHIE was derived. Thirty patients (63%) returned the SleepStrip, the device displayed an AHISS in 22 patients (73%), of which 17 (57%) was deemed valid by the device. Twelve of the 30 patients who returned the SleepStrip had an AHIE > or = 20 on the detailed sleep study. Of these, the SleepStrip recording was valid in seven of which only two had an AHISS > or = 20. We concluded that the SleepStrip was not suitable for unsupervised postal screening for OSA. PMID- 12753354 TI - Association of right ventricular dysfunction and Cheyne-Stokes respiration in patients with chronic heart failure. AB - Cheyne-Stokes respiration (CSR) is present in up to 40% of patients with congestive heart failure (CHF) and is an independent risk factor for increased overall mortality. We examined whether CSR is associated with right ventricular (RV) dysfunction in CHF patients. Parameters of RV function were assessed by two dimensional echocardiography and tissue velocity imaging in 42 patients (aged 23 75 years) with a left ventricular (LV) ejection fraction below 40%. Respiratory polygraphy revealed CSR with an central apnea-hypopnea index (CAHI) >10 h-1 in 13 of the 42 patients (31%). Demographic characteristics did not differ among the patient groups. The velocity of the tricuspid annular systolic motion (TASM), a parameter reflecting systolic RV function, was significantly reduced in CHF patients with CSR (10.5 +/- 2.3 cm s-1) compared with those without CSR (15.0 +/- 5.1 cm s-1, P = 0.004), and was inversely associated with the CAHI (y = 15.2 0.2x; r = 0.46, P = 0.003). The RV dimensions were significantly increased and the fractional RV area changes significantly reduced in CHF patients with CSR (33 +/- 17 versus 48 +/- 20%; P = 0.04). Doppler parameters of pulmonary artery flow indicate higher pulmonary artery pressures in CSR patients compared with patients without CSR, which is also reflected by an increased RV free-wall thickness in CSR patients (6.5 +/- 1.1 vs. 5.3 +/- 1.3 mm; P = 0.05). Parameters of systolic LV function, forced expiratory volume in 1 s (FEV1), and PaO2 and PaCO2 were not different among patients with or without CSR. In conclusion, CSR is associated with depressed systolic RV function and increased RV dimensions in CHF patients. Future studies will show whether optimized treatment of CSR will improve RV function. PMID- 12753355 TI - Effect of ondansetron on moderate obstructive sleep apnoea, a single night, placebo-controlled trial. PMID- 12753356 TI - The effect of social stimuli on the accumulation of sleep debt and need for recovery: does copulatory activity increase slow-wave sleep? PMID- 12753358 TI - Root surface debridement and endotoxin removal. AB - OBJECTIVE: This study assessed associations between the number of standardized scaling strokes and the reduction of endotoxin on the root surface. BACKGROUND: Therapy of periodontally involved teeth attempts removal of accretions by scaling and root planing. The amount of mechanical therapy required to free the root surface from etiologic factors remains unknown. METHODS: Twenty-four extracted human caries-free single rooted teeth with at least 5 mm attachment loss were used. A region of interest (ROI) which contained subgingival calculus was defined on the root surface of each tooth. Standardized force instrumentation was applied using a force-measuring curet. Fifty working strokes were applied to every ROI. Forces applied were recorded. The force recordings were converted from Millivolts into Newtons (N). After every unit of 5 strokes, presence of calculus was evaluated and scaling debris was collected. Endotoxin concentration was determined in the debris samples. RESULTS: The endotoxin values for strokes 1-5 were statistically significantly greater than the values from all other stroke intervals. Complete calculus removal occurred after a mean of 9.3 strokes. The endotoxin concentration reached a minimal level with concentrations of 0.03-0.306 EU/ml after calculus removal was complete. CONCLUSION: These findings suggest that completion of calculus removal coincides with endotoxin levels associated with clinically healthy teeth. PMID- 12753359 TI - Histomorphometric characteristics and expression of epidermal growth factor and its receptor by epithelial cells of normal gingiva and hereditary gingival fibromatosis. AB - OBJECTIVE: The objective of this study was to examine the histomorphometric features and evaluate the expression of epidermal growth factor (EGF) and transmembranic receptor (EGFr) and the proliferative potential of epithelial cells from normal and hereditary gingival fibromatosis (HGF) gingival tissues. BACKGROUND: EGF is a multifunctional cytokine with a variety of biological effects including stimulation of cell proliferation by binding to its specific EGFr. METHODS: Immunohistochemistry was performed to measure EGF and EGFr expression and the epithelial cell proliferation was determined by measuring proliferating cell nuclear antigen (PCNA). RESULTS: Histomorphometric evaluation indicated that in HGF the mean height of the epithelial papillae was higher compared to the normal gingiva (NG), whereas mean epithelial area and number of epithelial papillae were quite similar in both groups. The EGF and EGFr positive cells were observed in the basal, spinous and granular cell layers of both normal and HGF tissues, with a gradual reduction from the basal layer. Although the expressions of EGF and EGFr in the control group were significantly higher than those from HGF, in HGF the epithelial papilla tips showed increased number of proliferating cells and elevated expression of EGF and EGFr. There was a correlation between the proliferative potential of epithelial cells and the expression of EGF or EGFr only in the epithelial papilla tips of HGF gingiva. CONCLUSION: Our data suggest that EGF and EGFr in the oral epithelium of HGF gingiva may stimulate epithelial cell proliferation, with the resultant apical migration of the oral epithelium and formation of the slender deep epithelial papillae; however, without hyperplastic alterations. PMID- 12753360 TI - Expression and intracellular localization of progesterone receptors in cultured human gingival fibroblasts. AB - OBJECTIVE: The aim of this immunocytochemical study was to characterize the expression and distribution of the progesterone receptor (PR) and estrogen receptor (ER) in gingival fibroblasts using culture cells derived from people at various ages. BACKGROUND: The reaction of female hormones is tissue or cell specific, and receptor availability in the cell is one of the major causes for the different reactions. Gingiva is a target tissue for female hormones; however, the characteristics of PR and ER in both the fibroblasts and the other component cells remain largely unknown. MATERIALS: Gingival tissue was obtained from six people at various ages and culture fibroblasts were established. At least three passages of each cell line were strained for PR and ER with monoclonal antibodies (Clone 1A6, Clone 1D5, respectively). RESULTS: PR positive cells were detected in all six cell lines through early passages to late ones, but ER were only observed in two of six samples with faint reactions. The staining intensity for PR was greater than for ER, but less than that shown in the MCF-7 breast cancer cells, positive control. In every positive control test, ER reactivity was equal to or higher than that of PR. During the interphase, significantly fewer positive fibroblasts occurred compared with negative fibroblasts, and positive nuclei were even fewer. Meanwhile, most of the mitotic cells were PR positive, showing intense localization around chromosomes and on microtubules. These findings suggest that gingival fibroblasts are fundamentally capable of expressing PR and transmit the signal to target genes. CONCLUSIONS: The present study may conclude that in either gender or at any age, gingival fibroblasts express PR rather low in level and do not necessarily localize PR in a nuclear dominant fashion, which is an essential feature for reproductive organ cells. The poor ER reactivity shown in the gingival fibroblasts was discussed in view of the receptor subtype. PMID- 12753361 TI - Interleukin-1beta, clinical parameters and matched cellular-histopathologic changes of biopsied gingival tissue from periodontitis patients. AB - OBJECTIVE: The aim of the present study was to investigate whether interleukin (IL)-1beta in diseased tissues adjacent to periodontal pockets can reflect the degree of inflammation and destruction of these tissues pathologically. BACKGROUND: IL-1beta-dependent mechanisms have been strongly implicated in contributing to inflammation and destruction of bone and attachment loss, which are characteristic features of periodontal disease. This biochemical mediator released during pro-inflammatory processes has not been objectively integrated with clinical and histopathologic features of periodontal disease. METHODS: Periodontitis-affected inflamed tissue and clinically nonaffected healthy gingivae were harvested from 14 periodontal patients, respectively. The severity of tissue inflammation was illustrated by clinical parameters and cellular histologic changes and quantified by histometric assessments. IL-1beta in these extracted specimens was measured with an enzyme-linked immunosorbent assay (ELISA) technique. Pathogenic roles that IL-1beta plays in gingival inflammation and pathologic tissue changes in tissue sections were analyzed statistically. RESULTS: The overall total tissue IL-1beta, tissue concentration of IL-1beta, and percentage of inflammatory cell infiltration (PICI) determined from diseased gingivae were obviously higher than those of controls from both healthy sites of periodontitis and non-periodontitis subjects. With increasing gingival index (GI), plaque index (PlI), and probing depth (PD), there was a marked elevation in total tissue IL-1beta. Total tissue IL-1beta was significantly correlated with GI, PlI, the PICI, and tissue alterations. Polymorphonuclear leukocytes (PMNs) and monocyte-macrophage cells seemed to predominate in heavily infiltrated areas of diseased gingiva. These cell types were confirmed by immunocytochemical localization with either monoclonal mouse antihuman neutrophil elastase antibody or monoclonal mouse antihuman macrophage (CD68) antibody, respectively. Total tissue IL-1beta and the PICI were also elevated in diseased gingivae near deeper PD, while neither total IL-1beta nor tissue concentration was statistically correlated with PD. Thus, correlation analysis indicates that IL-1beta level in inflamed periodontal tissues correlates highly with clinical parameters (GI and PlI) and PICI (the degree of inflammation). CONCLUSIONS: These observations suggest that IL-1beta plays a significant role in the pathogenic mechanisms of periodontal tissue destruction, and that measurement of tissue IL-1beta would be a valuable aid and useful for diagnostic markers of periodontal diseases. PMID- 12753362 TI - Unique genes induced by mechanical stress in periodontal ligament cells. AB - OBJECTIVES: The aim of this study is to isolate mechanical stress-induced genes (MSGens) from human periodontal ligament (PDL) cells and to analyze profiles of the mRNA expression of these genes. BACKGROUND: Differential expression of genes in PDL cells under physiological stress such as occlusal force is thought to be orchestrated not only for the remodeling of PDL itself but also for the repair and regeneration of periodontal tissues. However, little is known about the genes expressed in PDL cells under mechanical stress. METHODS: The cDNA from mechanical stress-applied human PDL cells was subtracted against the cDNA from static control cells. The subtracted cDNA was amplified by polymerase chain reaction (PCR) and cloned for further analysis. RESULTS: Among 68 independent clones isolated, 15 contained DNA fragments greater than 250 bp. Reverse Northern analysis revealed a marked induction of MSGen-15 and MSGen-28 mRNA expression in the mechanical stress-applied cells. However, little difference in the magnitude of expression for the other MSGens was detected between the stress-applied cells and the control cells. After nucleotide sequencing and the analysis of homology with known genes, five clones were identified; ribosomal protein S27 (MSGen-9), MRG 15 (MSGen-15), androgen-binding protein (MSGen-18), cathepsin H (MSGen-28), and cytochrome c (MSGen-47). Interestingly, it has been reported that MRG 15 is a novel transcription factor involved in the regulation of cell growth and senescence. The remaining 10 clones, classified into six sequence types, had no significant homology with any known genes. CONCLUSIONS: These results suggest that many known and unknown genes are expressed in response to mechanical stress in PDL cells, and that a transcription factor, MRG 15, may be responsible for molecular events in PDL cells under mechanical stress. PMID- 12753363 TI - Generation of gingival T cell lines/clones specific with Porphyromonas gingivalis pulsed dendritic cells from periodontitis patients. AB - OBJECTIVES AND BACKGROUND: It is well documented that in periodontitis lesions, most infiltrated gingival T cells are antigen-specific memory T cells. These cells play an important role as regulators and effector cells in the pathogenesis of periodontitis. In this study, we used dendritic cells (DCs) as antigen presenting cells to generate human gingival T cell lines and clones specific for Porphyromonas gingivalis from periodontitis patients. METHODS: Autologous DCs were derived from the patients' adherent monocytes using granulocyte-macrophage colony-stimulating factor and interleukin (IL)-4. Lymphocytes were isolated from gingival biopsies using collagenase enzyme digestion and the number was increased by subsequent culturing in IL-2-containing medium. T cells were then negatively sorted using flow cytometry, cocultured with P. gingivalis-pulsed DCs and subsequently expanded in the culture medium containing IL-2. T cells were kept viable and active by periodic exposure to antigen-pulsed DCs. The specificity of the T cell lines was tested against four plaque bacteria: P. gingivalis, Actinobacillus actinomycetemcomitans, Prevotella intermedia and Actinomyces viscosus. The established T cell lines were then cloned. Three P. gingivalis specific T cell lines and 12 gingival T cell clones were generated. They all showed good specificity against P. gingivalis but not to other plaque bacteria. RESULTS: All T cell clones were positive for CD4 and the majority of them produced interferon gamma, but a minimal or negligible amount of IL-5. CONCLUSIONS: The data obtained clearly showed that monocyte-derived DCs could be used as powerful antigen-presenting cells to generate antigen-specific T cells from periodontitis tissues. PMID- 12753364 TI - Expression of endothelins and their receptors in cells from human periodontal tissues. AB - OBJECTIVE: The present study investigated the presence of ET-1 in gingival crevicular fluid (GCF) from patients with periodontitis, and the expression of endothelins (ETs) and their receptors mRNA in cultured cells from human periodontal tissues. BACKGROUND: ET was originally discovered as a potent vasoconstrictive peptide from endothelial cells. It has been reported that ETs are produced by various cells besides endothelial cells. ETs are related to inflammatory and sclerotic lesions, such as arteriolosclerosis and hepatic cirrhosis. Therefore, ETs may be involved in periodontal disease. However, the roles of ETs in development and progression of periodontal disease are not clear. METHODS: ET-1 released from the cultured cells was measured by enzyme-linked immunosorbent assay. mRNA expressions for ETs and their receptors were examined by reverse transcription-polymerase chain reaction and Northern blotting analysis. RESULTS: ET-1 levels in GCF from patients with periodontitis were higher than those from healthy subjects. Human gingival keratinocytes (HGK) expressed mRNA for ETs and their receptors, ET-Ar and ET-Br. ET-1 mRNA expression and ET-1 peptide production from HGK were enhanced by interleukin-1beta and tumor necrosis factor-alpha. CONCLUSIONS: These results suggest that ET-1 plays a significant role in periodontal disease. PMID- 12753365 TI - Serum antibody reactivity against recombinant PrtC of Porphyromonas gingivalis following periodontal therapy. AB - OBJECTIVES: In 34 patients with chronic periodontitis, the presence of IgA, IgG, and IgG subclass serum antibodies against recombinant PrtC (rPrtC) of Porphyromonas gingivalis was assessed by immunoblot analysis 24 months after therapy. METHODS: rPrtC was produced from P. gingivalis ATTC 33277 using the plasmid pGEX-2T. In addition, intraoral colonization with P. gingivalis was detected by PCR in subgingival plaque and swab samples from buccal mucosae, tonsils and tongue at baseline, 10 d, and 3, 6, 9, 12, 18, and 24 months. RESULTS: All patients were found to harbor P. gingivalis in the oral cavity at least once during the observation period. The identified antibody responses against the rPrtC of P. gingivalis were IgA (97%, i.e. 33/34 patients) and IgG (100%, i.e. 34/34), with an IgG subclass distribution of IgG2 (65%, i.e. 22/34 patients) > IgG3 (47%, i.e. 16/34) > IgG1 (38%, i.e. 13/34) > IgG4 (29%, i.e. 10/34). Anti-rPrtC IgA and IgG antibody reactivity was found in all but one patients (anti-rPrtC IgA negative), who tested negative for P. gingivalis at all of the assessed intraoral sites for at least 6 months before sera collection. There was no association between IgG subclass reactivity against the rPrtC of P. gingivalis and progression of periodontal attachment loss. CONCLUSION: The results indicated that anti-rPrtC IgA and IgG antibodies may serve as an indicator for past or present intraoral colonization with P. gingivalis. PMID- 12753366 TI - Inhibitory effect of procyanidin oligomer from elm cortex on the matrix metalloproteinases and proteases of periodontopathogens. AB - OBJECTIVES: The purpose of this study was to evaluate a partially purified extract (elm extract) from the Ulmi cortex (Ulmi macrocarpa Hance) and its active ingredient, a mix of procyanidin oligomers (3 to 12 flavan-3-ol monomers, an average molecular weight of 1,518 with an average polymerization degree of 5.3) for a possible inhibitory effect against proteases. BACKGROUND: Host-derived matrix metalloproteinases (MMPs) and bacterial proteases play important roles in the gingival tissue destruction that is a characteristic of periodontitis. The inhibitors of these proteases may be developed into therapeutic agents against periodontitis. METHODS: The inhibitory effects were assessed by gelatin zymography. The MMPs tested were originated from the gingival crevicular fluid (GCF) of adult periodontitis patients and from the conditioned media of cultured periodontal ligament (PDL) cells, which provided the proMMP-2 and activated MMP-2 when treated with a periodontopathogen, Treponema lecithinolyticum. Bacterial enzymes tested were secreted forms from two major periodontopathogens, Porphyromonas gingivalis and Treponema denticola. In addition, the inhibitory effects on trypsin-like enzymes from these two periodontopathogens were assayed by the n-benzoyl-DL-arginine-naphthylamide (BANA) test. RESULTS: The elm extract and the procyanidin oligomer (100-1,000 microg/ml) exhibited potent inhibitory effects on the MMPs in GCF (chiefly MMP-8 and MMP-9), the pro and active forms of MMP-2, and secreted and trypsin-like enzymes from T. denticola and P. gingivalis. CONCLUSIONS: These results suggest that elm cortex should be considered as a potential agent against periodontal diseases, due to its inhibitory action on MMPs and the proteases of periodontopathogens. PMID- 12753367 TI - Effect of Emdogain on human periodontal fibroblasts in an in vitro wound-healing model. AB - OBJECTIVE: The aim of this study was to evaluate the influence of Emdogain (EMD) on cultured gingival fibroblasts, periodontal ligament fibroblasts and dermal fibroblasts, using an in vitro model of wound healing. BACKGROUND: Enamel matrix derivative has been demonstrated to promote periodontal regeneration. However, the precise mechanisms by which this agent acts are still unclear. METHODS: The effect of EMD on proliferation of the cells was studied using subconfluent cultures of gingival fibroblasts and periodontal ligament fibroblasts. The cells were made quiescent overnight and then stimulated with various concentrations of EMD (10, 50, 100 and 150 microg/ml) for 24 h. Negative and positive controls were cells cultured in media containing 0.2% and 10% fetal calf serum (FCS). The DNA synthesis was measured by the cellular uptake of [3H]thymidine. For in vitro wounding the cells were cultured, wounded and stimulated with 0.2% FCS, 10% FCS and EMD at a concentration of 20 microg/ml. The percentage of wound fill after treatment was measured after d 1, 4, 6, 12 and 16. The proliferation of cells was also calculated by the extent of incorporation of crystal violet. RESULTS: The results demonstrated that cells in vitro fill an empty space by a combination of proliferation and cell migration. The most rapid closure of a wound area occurred where both proliferation and migration can occur as was seen when wounded cultures were maintained in 10% FCS or at a concentration of 20 microg/ml EMD which promoted proliferation. CONCLUSIONS: Therefore, EMD appears to exert an influence on cells that is compatible with improved wound healing. PMID- 12753368 TI - Effect of bone chip orientation on quantitative estimates of changes in bone mass using digital subtraction radiography. AB - OBJECTIVES: To assess the effect of the orientation of arbitrarily shaped bone chips on the correlation between radiographic estimates of bone loss and true mineral loss using digital subtraction radiography. METHODS: Twenty arbitrarily shaped bone chips (dry weight 1-10 mg) were placed individually on the superior lingual aspect of the interdental alveolar bone of a dry dentate hemi-mandible. After acquiring the first baseline image, each chip was rotated 90 degrees and a second radiograph was captured. Follow-up images were created without the bone chips and after rotating the mandible 0, 1, 2, 4, and 6 degrees around a vertical axis. Aluminum step tablet intensities were used to normalize image intensities for each image pair. Follow-up images were registered and geometrically standardized using projective standardization. Bone chips were dry ashed and analyzed for calcium content using atomic absorption. RESULTS: No significant difference was found between the radiographic estimates of bone loss from the different bone chip orientations (Wilcoxon: P > 0.05). The correlation between the two series of estimates for all rotations was 0.93 (Spearman: P < 0.05). Linear regression analysis indicated that both correlates did not differ appreciably ( and ). CONCLUSION: It is concluded that the spatial orientation of arbitrarily shaped bone chips does not have a significant impact on quantitative estimates of changes in bone mass in digital subtraction radiography. These results were obtained in the presence of irreversible projection errors of up to six degrees and after application of projective standardization for image reconstruction and image registration. PMID- 12753369 TI - Granulocyte elastase activity in static and flow gingival crevicular fluid. AB - OBJECTIVES: This study aimed to evaluate the volume of gingival crevicular fluid (GCF) and granulocyte elastase activity in static GCF (sGCF) and flow GCF (fGCF) from subjects with various periodontal conditions. METHODS: Eleven periodontally healthy, 10 gingivitis and 12 periodontitis subjects were recruited and the sites investigated consisted of healthy sites from healthy subjects (HH); healthy (HG) and gingivitis sites (GG) from gingivitis subjects; and healthy (HP), gingivitis (GP) and periodontitis sites (PP) from periodontitis subjects. fGCF samples were collected either 1 min or 5 min following sGCF collection by paper strip technique. GCF volume was determined by Periotron 6000 and granulocyte elastase activity was assayed with a specific substrate [l-pyroglutamyl-l-prolyl-l-valine p-nitroanilide(pGluProVal-pNA)]. RESULTS: At baseline, no significant differences existed in clinical and GCF parameters between the two matched sites for subsequent collection of fGCF samples either 1 min or 5 min after sGCF sampling in all subjects. The flow exudate in HG and HP sites quickly replenished to sGCF levels, while a delayed replenishment was found in HH sites, despite the similar sGCF volumes of these sites. The GCF volume and elastase levels in the fGCF at 1 min were higher in GP sites than in GG sites (P < 0.05). Overall, depletion of elastase levels in the fGCF at 1 min was observed in all subjects, whereas elastase levels in the fGCF at 5 min had replenished to sGCF levels in HP, GP, PP sites and GG sites, but had remained at a lower level in HH and HG sites. An overall positive correlation was found between sGCF and fGCF for GCF volume and elastase activity (P < 0.001); however, this correlation varied with GCF parameters and with site conditions of the subjects concerned. CONCLUSIONS: This study shows that patterns of dynamic changes in GCF flow and elastase activity varied under different periodontal conditions. Assessment of both sGCF and fGCF may allow better insight into the dynamic change of the target components in GCF. PMID- 12753370 TI - Povidone-iodine as a periodontal pocket disinfectant. AB - OBJECTIVES AND BACKGROUND: Povidone-iodine [polyvinylpyrrolidone-iodine complex (PVP-iodine)] might constitute a valuable adjunct to current periodontal therapy because of its broad-spectrum antimicrobial activity, low potential for developing resistance and adverse reactions, wide availability, ease of use, and low financial cost. This investigation employed a randomized, split-mouth study design to determine the microbiological and clinical effects of 10% PVP-iodine subgingival irrigation in periodontitis lesions showing radiographic evidence of subgingival calculus. METHODS: Sixteen adults having at least one periodontal pocket of 6 mm or more in each quadrant of the dentition and harboring one or more periodontopathic bacteria participated in the study. In each subject, a study site in each quadrant was randomly chosen to receive either subgingival irrigation with 10% PVP-iodine together with scaling and root planing, scaling and root planing alone, subgingival irrigation with 10% PVP-iodine, or subgingival irrigation with sterile saline. Prior to therapy and at 5 weeks post treatment, microbiological culture was carried out without knowledge of the clinical status or the type of treatment rendered. A blinded clinical examiner determined presence of dental plaque, probing pocket depth, and gingival bleeding on probing. Microbiological and clinical data were analyzed using a repeated measures analysis of variance and Kruskal-Wallis rank test with the Tukey and Mann-Whitney post hoc tests. RESULTS: At 5 weeks post-treatment, subgingival irrigation with PVP-iodine together with scaling and root planing caused a 95% or greater reduction in total pathogen counts in 44% of pockets having >/= 6 mm depth whereas scaling and root planing alone, povidone-iodine irrigation alone and water irrigation alone caused 95% reduction of total pathogens only in 6-13% of similar study sites (P = 0.02). Reduction in mean pocket depth was 1.8 mm for the PVP-iodine/scaling and root planing group, 1.6 mm for the scaling and root planing group, and 0.9 mm for the PVP-iodine and the saline monotherapy groups, with statistical significance reached for the scaling and root planing group vs. the PVP-iodine group (P = 0.04) and for the scaling and root planing group vs. the saline group (P = 0.02). Reduction in visible dental plaque, which ranged from 38% to 62%, showed no significant differences among treatment groups. CONCLUSIONS: The addition of subgingival PVP-iodine irrigation to conventional mechanical therapy may be a cost-effective means of reducing total counts of periodontal pathogens and helping control periodontal disease. However, subgingival irrigation with PVP-iodine without concomitant mechanical debridement might not improve microbiological and clinical variables in comparison with saline irrigation, at least not in sites with radiographic evidence of subgingival calculus. PMID- 12753371 TI - The herpesvirus-Porphyromonas gingivalis-periodontitis axis. AB - OBJECTIVES AND BACKGROUND: Members of the herpesvirus family have accumulated considerable support for a role in severe types of periodontitis. This study aimed to examine whether human cytomegalovirus (HCMV), Epstein-Barr virus type 1 (EBV-1) or herpes simplex virus (HSV) together with the major periodontopathic bacterium Porphyromonas gingivalis might interact in the pathogenesis of periodontal breakdown. METHODS: Sixteen subjects each contributed paper point samples from two progressing and two stable periodontitis lesions, as determined by ongoing loss of probing attachment. Polymerase chain reaction methodology was used to identify subgingival herpesviruses, P. gingivalis and other bacterial pathogens. Chi-squared tests and multivariate logistic regression were employed to identify statistical associations between herpesviruses, periodontopathic bacteria and clinical variables. RESULTS: HCMV and HSV were both significant predictors of the presence of subgingival P. gingivalis. In turn, P. gingivalis was positively associated with periodontitis active disease, probing attachment level, probing pocket depth, gingival bleeding upon probing and patient age. EBV 1 was not linked to P. gingivalis, although the virus was predictive of periodontitis active disease. The periodontitis disease risk associated with herpesvirus-P. gingivalis combinations depended on both site-specific and subject specific factors. CONCLUSION: The present data of aggressive periodontitis implicate HCMV, HSV and P. gingivalis as either cofactors in its etiology or triggers of relapses. Further studies are needed to determine the spectrum of periodontopathogenicity of herpesviruses and effective management of these viruses in periodontal sites. PMID- 12753372 TI - Effect of soluble receptors to interleukin-1 and tumor necrosis factor alpha on experimentally induced root resorption in rats. AB - OBJECTIVE: In this study, the role of the inflammatory cytokines interleukin-1 (IL-1) and tumor necrosis factor alpha (TNFalpha) in the course of mechanically induced root resorption was investigated. METHODS: Mechanical induction of root resorption was performed on the upper left first molars in 18 male Wistar rats according to the method of Nakane and Kameyama. Starting on day minus 1, six animals received daily intraperitoneal injections of 2 ml of 1 micro g/ml soluble receptors to IL-1 (sIL-1RII) and another six animals were administered the same dose of soluble receptors to TNFalpha (sTNFalpha-RI). Six animals served as a control. On d 7 the left maxillae were prepared for histological and morphometric analysis of the extent of the root resorption that had developed. RESULTS: The qualitative and quantitative results demonstrated that in both receptor groups the amount of root resorption was significantly reduced. Especially following systemic application of sTNFalpha-RI, root resorption was nearly completely prevented. CONCLUSIONS: Our results indicate that IL-1 and more particularly TNFalpha are important for the induction and the further process of mechanically induced root resorption in the rat. PMID- 12753374 TI - Calcification of degenerating tissues in the periodontal ligament during tooth movement. AB - OBJECTIVE: Calcification of degenerating tissues in the periodontal ligament (PDL) during tooth movement was investigated longitudinally. MATERIALS AND METHODS: Upper first molars of male Wistar rats were moved lingually for 1, 7 and 21 d, following which unfixed undecalcified sections of the lingual PDL (in the pressure zone) were examined histologically, histochemically (autoradiography and electron probe microanalysis). RESULTS: On d 1 of tooth movement, degenerating tissues, together with some calcified particles, were visible in the pressure zone of the lingual PDL. On d 7, substantial calcified aggregations were seen in the degenerating tissues, predominantly situated between the bone and root. This was confirmed by the 45Ca autoradiography. On d 21 of tooth movement, large calcified aggregations were still clearly evident between the bone and root. CONCLUSIONS: This calcification of the degenerating tissues is a self-defense response of the living body to prevent direct contact between alveolar bone and the tooth root during compression of the PDL, so preventing friction between them and the development of ankylosis. PMID- 12753373 TI - The effects of enamel matrix derivative (EMD) on osteoblastic cells in culture and bone regeneration in a rat skull defect. AB - OBJECTIVE: Enamel matrix derivative (EMD) has been clinically used to promote periodontal tissue regeneration. The purpose of the present study is to clarify EMD affects on osteoblastic cells and bone regeneration. MATERIALS AND METHODS: Mouse osteoblastic cells (ST2 cells and KUSA/A1 cells) are used in culture experiments. After cells were treated with EMD, cell growth was evaluated with DNA measurement, 5-bromo-2'-deoxyurydine (BrdU) incorporation assay. Measurement of alkaline phosphatase (ALP) activity and mineralized-nodule (MN) formation, Northern blotting analysis and zymography are also performed. In addition, EMD was applied to a rat skull defect and the defect was radiographically and histologically evaluated 2 weeks after the application. RESULTS: EMD did not stimulate ST2 cell growth; however, it enhanced KUSA/A1 cell proliferation. Although EMD stimulated ALP activity in both the cells, ALP activity in KUSA/A1 cells was affected to a much greater degree. Corresponding to the increase in ALP activity, MN formation in KUSA/A1 cells was enhanced by EMD. EMD stimulated osteoblastic phenotype expression of KUSA/A1 cells such as type I collagen, osteopontin, transforming growth factor beta 1 and osteocalcin. EMD treatment also stimulated matrix metalloproteinase production in KUSA/A1 cells. Although the effects of EMD on osteoblastic cells depend on cell type, the overall effect of EMD on osteoblastic cells is stimulatory rather than inhibitory. Finally, EMD application to a rat skull defect accelerated new bone formation. CONCLUSION: These results indicate that EMD affects osteoblastic cells and has potential as a therapeutic material for bone healing. PMID- 12753375 TI - Stress may enhance nicotine effects on periodontal tissues. An in vivo study in rats. AB - OBJECTIVES AND BACKGROUND: Diabetes mellitus and smoking have been described as important risk factors that may affect the initiation and progression of periodontitis. Recent studies have pointed to potentially periodontal risk indicators, which include stress. The present study investigated the effects of stress associated with nicotine administration on periodontal breakdown resulting from ligature-induced periodontitis in rats. METHODS: Twenty adult male Wistar rats were used. After anesthesia, both mandibular first molars received a cotton ligature in the dento-gingival area. The animals were randomly assigned to one of the following experimental groups: A - saline solution, B - 0.73 mg of nicotine/kg/d (intraperitoneal), C - stress (immobilization - 2 h/d/40 d) associated with an intraperitoneal administration of saline solution, and D - stress (immobilization - 2 h/d/40 d) associated with an intraperitoneal injection of 0.73 mg of nicotine/kg/d. Forty days later, the animals were sacrificed and the specimens routinely processed for serial decalcified sections. RESULTS: Intergroup analysis (ANOVA) revealed a greater bone loss (P < 0.05) in the animals of group D compared with the animals from groups A, B and C. In addition, the data revealed a significant effect of nicotine (group B) compared with groups A and C (P < 0.05), and no difference between groups A and C (P > 0.05). CONCLUSION: Within the limits of the present study, although stress did not affect periodontitis by itself, it significantly enhanced the effects of nicotine on the periodontal tissues. PMID- 12753376 TI - Effect of peptide conformation on membrane permeability. AB - The effect of peptide conformational constraint on the peptide permeation across the model membranes was examined by determining the permeability of pairs of cyclic and acyclic peptides related to c[d-Pen2, d-Pen5] enkephalin (DPDPE). The peptides were cyclized by formation of an intramolecular disulfide bridge between the second and fifth residues composed of either d-penicillamine or cysteine. In each case the acyclic peptide was three to seven times more permeable than corresponding cyclic peptide. The possibility that the differences in permeability of cyclic and acyclic peptides is based on the greater conformational freedom of the acyclic peptides in the presence of membrane was examined in more detail by isothermal titration calorimetric studies of Trp6 DPDPE and its acyclic analog. The membrane binding of the acyclic peptide is a more exothermic process than binding of its cyclic Trp6-DPDPE. The transfer of acyclic peptide from water to membrane is an enthalpy driven process, whereas the transfer of the cyclic peptide is driven by entropy. PMID- 12753377 TI - Solid-phase synthesis of polymyxin B1 and analogues via a safety-catch approach. AB - As part of a program towards the development of novel antibiotics, a convenient method for solid-phase synthesis of the cyclic cationic peptide polymyxin B1 and analogues thereof is described. The methodology, based on cleavage-by-cyclization using Kenner's safety-catch linker, yields crude products with purities ranging from 37-67%. Antibacterial assays revealed that analogues 23-26, in which the (S) 6-methyloctanoic acid moiety is replaced with shorter acyl chains, exhibit distinct antimicrobial activity. The results suggest that the length of the acyl chain is rather critical for antimicrobial activity. On the other hand, substitution of the hydrophobic ring-segment D-Phe-6/Leu-7 in polymyxin B1 with dipeptide mimics (i.e. analogues 27-33) resulted in almost complete loss of antimicrobial activity. PMID- 12753378 TI - Direct binding properties of conantokins to native N-methyl-d-aspartate receptors. AB - Conantokin-G (con-G) is a small, gamma-carboxyglutamic acid (Gla)-containing peptide that functions neurophysiologically by inhibiting the N-methyl-d aspartate receptor (NMDAR). In the current study, the receptor binding properties of an alanine-rich, Gla-deficient con-G variant, Ala-con-G, were assessed following tracer radioiodination with 125I. Direct binding experiments with [125I]Ala-con-G yielded a single site defined by a Kd value of 516 +/- 120 nm. Displacement of [125I]Ala-con-G binding by Ala-con-G resulted in 100% displacement with an IC50 value of 564 +/- 33 nm, while heterologous displacement by con-G[S16Y], con-G, con-T, and con-R[1-17] yielded IC50 values in the range of 15-45 microm. No displacement was observed with d-gamma-con-G or con-G[L5A], analogs that are inactive at NMDARs. Specific [125I]Ala-con-G binding was displaced by NMDA and 2-amino-5-phosphopentanoic acid in a dose-dependent manner, suggesting an interaction at the glutamate binding site. The direct binding of [125I]Ala-con-G to adult rat brain sections revealed an anatomical distribution of binding sites in all regions known to contain the NR2B subunit of the NMDAR. These results constitute the only known demonstration of the direct binding of a radiolabeled conantokin to the NMDARs present in rat brain membrane preparations and rat brain sections, and suggest that radiolabeled Ala-con-G, and similar conantokin derivatives, may find utility as probes of NMDARs in a variety of systems. PMID- 12753379 TI - Lipophilic derivatization of synthetic peptides belonging to NS3 and E2 proteins of GB virus-C (hepatitis G virus) and its effect on the interaction with model lipid membranes. AB - The synthesis by solid-phase methodologies of peptides belonging to structural and non-structural proteins of GB virus C as well as its N-alpha-acylation with myristate and palmitate fatty acids is described. To explore the peptide-lipid interactions we have used liposomes composed of dipalmitoylphosphatidylcholine as model membranes and complementary spectroscopic and calorimetric techniques. Our results show that structural and more clearly the structural lipophilic peptide sequences incorporated into lipid bilayers perturb the packing of lipids and affect their thermotropic properties, more than the non-structural selected sequence. However, the binding of the synthetic sequences to lipid membranes occurred without any restructuration of the peptides. PMID- 12753380 TI - Syntheses of cyclic prodrugs of RGD peptidomimetics with various macrocyclic ring sizes: evaluation of physicochemical, transport and antithrombic properties. AB - The objective of this work was to synthesize cyclic prodrugs 1a-d of RGD peptidomimetics 2a-d with various ring sizes (n[CH2] = 1, 3, 5 and 7) and to evaluate the effect of ring size on their transport, physicochemical, enzymatic stability, and antithrombic properties. The syntheses of cyclic prodrugs 1a-d were achieved by converging two key intermediates, Boc-Phe-O-CH2-OCO-OpNP (5) and H2N-(CH2)n-CO-Asp(OBzl)-OTce (8a-d), to give linear precursors Boc-Phe-O-CH2-OCO HN-(CH2)n-CO-Asp(OBzl)-OTce (9a-d). The N- and C-terminus protecting groups were removed from 9a-d to give 10a-d. Linear precursors 10a-d were cyclized, and the remaining Bzl-protecting group was removed to produce cyclic prodrugs 1a-d in around 20% overall yield. The linear RGD peptidomimetics (2a-d) were synthesized using standard Boc-amino acid chemistry by solution-phase method. Increasing the ring size by adding methylene groups also increases the hydrophobicity of the cyclic prodrugs and parent RGD peptidomimetics. The transport properties of cyclic prodrugs 1c and 1d were 2.6- and 4.4-fold better than those of parent compounds 2c and 2d, respectively. These results suggest that increasing the hydrophobicity of the cyclic prodrugs and parent RGD peptidomimetics enhanced their transport properties. The hydrodynamic radii of the cyclic prodrugs were also smaller than those of their respective parent compounds, suggesting that the change in size may contribute to their transport properties. The chemical stability of the cyclic prodrugs was affected by the ring size, and the cyclic prodrug with the larger ring size (i.e. 1d) was more stable than the smaller one (i.e. 1a). All the cyclic prodrugs were more stable at pH 4 than at pH 7 and 10. Prodrug-to-drug conversion could be induced by isolated esterase as well as esterase found in human plasma. An increase in the length of methylene group (n[CH2] = 1, 3, 5, 7) enhanced the antithrombic activity of the prodrugs and the parent compounds. In summary, the ring size of cyclic prodrugs affected their transport, physicochemical, and antithrombic properties. PMID- 12753383 TI - The pteridine pathway in zebrafish: regulation and specification during the determination of neural crest cell-fate. AB - This review describes pteridine biosynthesis and its relation to the differentiation of neural crest derivatives in zebrafish. During the embryonic development of these fish, neural crest precursor cells segregate into neural elements, ectomesenchymal cells and pigment cells; the latter then diversifying into melanophores, iridophores and xanthophores. The differentiation of neural cells, melanophores, and xanthophores is coupled closely with the onset of pteridine synthesis which starts from GTP and is regulated through the control of GTP cyclohydrolase I activity. De novo pteridine synthesis in embryos of this species increases during the first 72-h postfertilization, producing H4biopterin, which serves as a cofactor for neurotransmitter synthesis in neural cells and for tyrosine production in melanophores. Thereafter, sepiapterin (6-lactoyl-7,8 dihydropterin) accumulates as yellow pigment in xanthophores, together with 7 oxobiopterin, isoxanthopterin and 2,4,7-trioxopteridine. Sepiapterin is the key intermediate in the formation of 7-oxopteridines, which depends on the availability of enzymes belonging to the xanthine oxidoreductase family. Expression of the GTP cyclohydrolase I gene (gch) is found in neural cells, in melanoblasts and in early xanthophores (xanthoblasts) of early zebrafish embryos but steeply declines in xanthophores by 42-h postfertilization. The mechanism(s) whereby sepiapterin branches off from the GTP-H4biopterin pathway is currently unknown and will require further study. The surge of interest in zebrafish as a model for vertebrate development and its amenability to genetic manipulation provide powerful tools for analysing the functional commitment of neural crest derived cells and the regulation of pteridine synthesis in mammals. PMID- 12753384 TI - Serial analysis of gene expression (SAGE): advances, analysis and applications to pigment cell research. AB - As cells progress from normal to diseased states, they may undergo a series of gene expression changes. Advances in molecular biology allow us to examine a host of these changes at once, in a high throughput fashion. Serial analysis of gene expression (SAGE) allows for the expression profiling of the complete transcriptome of a given cell, and has the potential for identifying novel genes as well as those in low abundance. In this review, we will outline the technique, how one analyzes the massive amounts of data generated, and describe pigment cell libraries currently in the making. PMID- 12753385 TI - The inhibitory effect of androgen and sex-hormone-binding globulin on the intracellular cAMP level and tyrosinase activity of normal human melanocytes. AB - The effect of androgens on human melanocytes has not been well clarified. We studied the effects of androgens on normal human melanocytes in the presence or absence of sex-hormone-binding globulin (SHBG), which complexes with those hormones. Immunohistochemically, testosterone and SHBG co-localized on the cell membrane. Androgens such as testosterone, 5alpha-dihydrotestosterone, and methyltrienolone (R1881, a potent synthetic androgen), reduced intracellular cAMP levels after treatment with SHBG, but hydrocortisone had no effect. We also found that testosterone and R1881 slightly suppressed tyrosinase activity in melanocytes when treated with SHBG, although they had no effect on the expression of tyrosinase at the transcriptional or translational level, as measured by semi quantitative reverse transcriptase-polymerase chain reaction and by Western blot analysis, respectively. Our results suggest that androgens may modulate tyrosinase activity at the posttranslational level through the cell membrane signaling pathway. PMID- 12753386 TI - Screening of human primary melanocytes of defined melanocortin-1 receptor genotype: pigmentation marker, ultrastructural and UV-survival studies. AB - Recent population studies have demonstrated an association with the red-hair and fair-skin phenotype with variant alleles of the melanocortin-1 receptor (MC1R) which result in amino acid substitutions within the coding region leading to an altered receptor activity. In particular, Arg151Cys, Arg160Trp and Asp294His were the most commonly associated variants seen in the south-east Queensland population with at least one of these alleles found in 93% of those with red hair. In order to study the individual effects of these variants on melanocyte biology and melanocytic pigmentation, we established a series of human melanocyte strains genotyped for the MC1R receptor which included wild-type consensus, variant heterozygotes, compound heterozygotes and homozygotes for Arg151Cys, Arg160Trp, Val60Leu and Val92Met alleles. These strains ranged from darkly pigmented to amelanotic, with all strains of consensus sequence having dark pigmentation. UV sensitivity was found not to be associated with either MC1R genotype or the level of pigmentation with a range of sensitivities seen across all genotypes. Ultrastructural analysis demonstrated that while consensus strains contained stage IV melanosomes in their terminal dendrites, Arg151Cys and Arg160Trp homozygote strains contained only stage II melanosomes. This was despite being able to show expression of tyrosinase and tyrosinase-related protein-1 markers, although at reduced levels and an ability to convert exogenous 3,4-dihydroxyphenyl-alanine (DOPA) to melanin in these strains. PMID- 12753387 TI - Epidemiology of vitiligo and associated autoimmune diseases in Caucasian probands and their families. AB - Generalized vitiligo is an autoimmune disorder characterized by acquired white patches of skin and overlying hair, the result of loss of melanocytes from involved areas. The most common disorder of pigmentation, vitiligo occurs with a frequency of 0.1-2.0% in various populations. Family clustering of cases is not uncommon, in a non-Mendelian pattern suggestive of multifactorial, polygenic inheritance. We surveyed 2624 vitiligo probands from North America and the UK regarding clinical characteristics, familial involvement, and association with other autoimmune disorders, the largest such survey ever performed. More than 83% of probands were Caucasians, and the frequency of vitiligo appeared approximately equal in males and females. The frequency of vitiligo in probands' siblings was 6.1%, about 18 times the population frequency, suggesting a major genetic component in disease pathogenesis. Nevertheless, the concordance of vitiligo in monozygotic twins was only 23%, indicating that a non-genetic component also plays an important role. Probands with earlier disease onset tended to have more relatives affected with vitiligo, suggesting a greater genetic component in early onset families. The frequencies of six autoimmune disorders were significantly elevated in vitiligo probands and their first-degree relatives: vitiligo itself, autoimmune thyroid disease (particularly hypothyroidism), pernicious anaemia, Addison's disease, systemic lupus erythematosus, and probably inflammatory bowel disease. These associations indicate that vitiligo shares common genetic aetiologic links with these other autoimmune disorders. These results suggest that genomic analysis of families with generalized vitiligo and this specific constellation of associated autoimmune disorders will be important to identify the mechanisms of genetic susceptibility to autoimmunity. PMID- 12753388 TI - Regulation of melanosome movement by MAP kinase. AB - Our objectives were to further characterize the signaling pathways in melatonin induced aggregation in Xenopus melanophores, specifically to investigate a possible role of mitogen-activated protein kinase (MAPK). By Western blotting we found that melatonin activates MAPK, which precedes melanosome aggregation measured in a microplate reader. Activation of MAPK, tyrosine phosphorylation of a previously described 280-kDa protein, and melanosome aggregation are sensitive to PD98059, a selective inhibitor of MAPK kinase. The MAPK activation is also decreased by the adenylate cyclase stimulant forskolin. In summary, we found that MAPK is activated during melatonin-induced melanosome aggregation. Activation was decreased by an inhibitor of MAPK kinase, and by forskolin. In addition to inhibition of cyclic adenosine 3',5'-monophosphate (cAMP), reduction in protein kinase A activity (PKA), and activation of protein phosphatase 2A, we suggest that melatonin receptors activate the MAPK cascade and tyrosine phosphorylation of the 280-kDa protein. Although the cAMP/PKA signaling pathway is the most prominent, our data suggest that simultaneous activation of the MAPK cascade is of importance to obtain a completely aggregated state. This new regulatory mechanism of organelle transport by the MAPK cascade might be important in other eukaryotic cells. PMID- 12753395 TI - The IFPCS presidential lecture: a chemist's view of melanogenesis. AB - The significance of our understanding of the chemistry of melanin and melanogenesis is reviewed. Melanogenesis begins with the production of dopaquinone, a highly reactive o-quinone. Pulse radiolysis is a powerful tool to study the fates of such highly reactive melanin precursors. Based on pulse radiolysis data reported by Land et al. (J Photochem Photobiol B: Biol 2001;64:123) and our biochemical studies, a pathway for mixed melanogenesis is proposed. Melanogenesis proceeds in three distinctive steps. The initial step is the production of cysteinyldopas by the rapid addition of cysteine to dopaquinone, which continues as long as cysteine is present (1 microM). The second step is the oxidation of cysteinyldopas to give pheomelanin, which continues as long as cysteinyldopas are present (10 microM). The last step is the production of eumelanin, which begins only after most cysteinyldopas are depleted. It thus appears that eumelanin is deposited on the preformed pheomelanin and that the ratio of eu- to pheomelanin is determined by the tyrosinase activity and cysteine concentration. In eumelanogenesis, dopachrome is a rather stable molecule and spontaneously decomposes to give mostly 5,6 dihydroxyindole. Dopachrome tautomerase (Dct) catalyses the tautomerization of dopachrome to give mostly 5,6-dihydroxyindole-2-carboxylic acid (DHICA). Our study confirmed that the role of Dct is to increase the ratio of DHICA in eumelanin and to increase the production of eumelanin. In addition, the cytotoxicity of o-quinone melanin precursors was found to correlate with binding to proteins through the cysteine residues. Finally, it is still unknown how the availability of cysteine is controlled within the melanosome. PMID- 12753396 TI - The Seiji memorial lecture: the melanosome: an ideal model to study cellular differentiation. AB - Melanosomes provide an intriguing model for study at many levels. In part this is due to their unique structure and function, but also in part to their involvement in pigmentary diseases and as a model to study basic cellular mechanisms of organelle biogenesis. Recent studies have elucidated the full proteome of the melanosome and the metabolic and molecular lesions involved in a number of pigmentary diseases have been resolved. This paper summarizes recent advances in the field in these areas. PMID- 12753397 TI - DNA microarrays and likelihood ratio bioinformatic methods: discovery of human melanocyte biomarkers. AB - In this article, some of the advantages and limitations of DNA microarray technologies for gene expression profiling are summarized. As a model experiment, DermArray DNA microarrays were utilized to identify potential biomarkers of cultured normal human melanocytes in two different experimental comparisons. In the first case, melanocyte RNA was compared with vastly dissimilar non melanocytic RNA samples of normal skin keratinocytes and fibroblasts. In the second case, melanocyte RNA was compared with a primary cutaneous melanoma line (MS7) and a metastatic melanoma cell line (SKMel-28). The alternative approaches provide dramatically different lists of 'normal melanocyte' biomarkers. The most robust biomarkers were identified using principal component analysis bioinformatic methods related to likelihood ratios. Only three of 25 robust biomarkers in the melanocyte-proximal study (i.e. melanocytes vs. melanoma cells) were coincidentally identified in the melanocyte-distal study (i.e. melanocytes vs. non-melanocytic cells). Selected up-regulated biomarkers of melanocytes (i.e. TRP-1, melan-A/MART-1, silver/Pmel17, and nidogen-2) were validated by qRT-PCR. Some of the melanocytic biomarkers identified here may be useful in molecular diagnostics, as potential molecular targets for drug discovery, and for understanding the biochemistry of melanocytic cells. PMID- 12753398 TI - Melanocyte-specific immune response in melanoma and vitiligo: two faces of the same coin? AB - The appearance of depigmentation during the course of malignant melanoma has been considered a good prognostic sign. Is it only a side-effect, informative of the immune system's response to the treatment, or does it act as a necessary amplifier of these clinically important anti-tumor responses? The current review will attempt to tackle this question by reviewing the current literature, as well as by posing some novel hypotheses. Understanding the nature of humoral and cellular immune responses directed against normal melanocytes and their malignant counterparts may lead to the design of improved therapeutic strategies relevant to both vitiligo and melanoma. PMID- 12753399 TI - Microphthalmia-associated transcription factor in the Wnt signaling pathway. AB - Microphthalmia-associated transcription factor (MITF) contains a basic helix-loop helix and leucine-zipper (bHLH-LZ) structure and consists of many isoforms with different N-termini. Melanocyte-specific MITF isoform (MITF-M) is of particular interest, because a heterozygous mutation in the MITF gene is associated with Waardenburg syndrome type 2 (WS2) that is characterized by deafness and hypopigmentation because of lack of melanocytes in the inner ear and skin. Expression of MITF-M is under the regulation of the melanocyte-specific promoter (M promoter) of the MITF gene, and transcription from the M promoter is induced by Wnt signals through a nuclear mediator, lymphoid-enhancing factor 1 (LEF-1). In addition, functional cooperation of MITF-M with LEF-1 could lead to transcriptional activation of the M promoter and the dopachrome tautomerase (DCT) gene, an early melanoblast marker. The bHLH-LZ region of MITF-M is responsible for the physical interaction with LEF-1, and beta-catenin is required for the collaboration between LEF-1 and MITF-M. Importantly, MITF-M could function as a non-DNA-binding co-factor for LEF-1. These results suggest that MITF-M may function as a self-regulator of its own expression to maintain a threshold level of MITF-M at a certain sensitive stage of melanocyte development, which could account for the dominant inheritance of WS2. MITF-M therefore plays dual roles in the Wnt signaling pathway; MITF-M represents a downstream target and a nuclear mediator of Wnt signals in melanocytes. PMID- 12753400 TI - The role of melanocortin-1 receptor polymorphism in skin cancer risk phenotypes. AB - We have examined melanocortin-1 receptor (MC1R) variant allele frequencies in the general population and in a collection of adolescent dizygotic and monozygotic twins to determine statistical associations of pigmentation phenotypes with increased skin cancer risk. This included hair and skin color, freckling, mole count and sun exposed skin reflectance. Nine variants were studied and designated as either strong R (OR = 63; 95% CI 32-140) or weak r (OR = 5; 95% CI 3-11) red hair alleles. Penetrance of each MC1R variant allele was consistent with an allelic model where effects were multiplicative for red hair but additive for skin reflectance. To assess the interaction of the brown eye color gene BEY2/OCA2 on the phenotypic effects of variant MC1R alleles we imputed OCA2 genotype in the twin collection. A modifying effect of OCA2 on MC1R variant alleles was seen on constitutive skin color, freckling and mole count. In order to study the individual effects of these variants on pigmentation phenotype we have established a series of human primary melanocyte strains genotyped for the MC1R receptor. These include strains which are MC1R wild-type consensus, variant heterozygotes, and homozygotes for strong R alleles Arg151Cys and Arg160Trp. Ultrastructural analysis demonstrated that only consensus strains contained stage III and IV melanosomes in their terminal dendrites whereas Arg151Cys and Arg160Trp homozygous strains contained only immature stage I and II melanosomes. Such genetic association studies combined with the functional analysis of MC1R variant alleles in melanocytic cells should provide a link in understanding the association between pigmentary phototypes and skin cancer risk. PMID- 12753401 TI - Melanin as a target for melanoma chemotherapy: pro-oxidant effect of oxygen and metals on melanoma viability. AB - Melanoma cells have a poor ability to mediate oxidative stress, which may be attributed to constitutive abnormalities in their melanosomes. We hypothesize that disorganization of the melanosomes will allow chemical targeting of the melanin within. Chemical studies show that under oxidative conditions, synthetic melanins demonstrate increased metal affinity and a susceptibility to redox cycling with oxygen to form reactive oxygen species. The electron paramagnetic resonance (EPR)-active 5,5'-dimethyl-pyrollidine N-oxide spin adduct was used to show that binding of divalent Zn or Cu to melanin induces a pro-oxidant response under oxygen, generating superoxide and hydroxyl radicals. A similar pro-oxidant behaviour is seen in melanoma cell lines under external peroxide stress. Melanoma cultures grown under 95% O2/5% CO2 atmospheres show markedly reduced viability as compared with normal melanocytes. Cu- and Zn-dithiocarbamate complexes, which induce passive uptake of the metal ions into cells, show significant antimelanoma activity. The antimelanoma effect of metal- and oxygen-induced stress appears additive rather than synergistic; both treatments are shown to be significantly less toxic to melanocytes. PMID- 12753402 TI - Melanosome degradation: fact or fiction. AB - Our mini review summarizes what is known about the (bio)degradation of melanosomes. Unlike melanosome biogenesis where our knowledge enables us to explain it in molecular terms posing many interesting questions on the relation between lysosomes and melanosomes, melanosome degradation has remained 'terra incognita'. Observations at optical and ultrastructural levels describe the disintegration of melanosomes in the lysosomal compartment (in auto- and heterophagosomes). Histochemical studies suggest the participation of acid hydrolases in the process of melanosome degradation. Biochemical data confirm the ability of lysosomal hydrolases to degrade melanosome constituents except the melanin moiety. The similarity of melanin structure to that of polycyclic aromatic hydrocarbons suggests that melanin should be sensitive mainly, if not exclusively, to oxidative breakdown. In vitro melanin can indeed be decomposed by an oxidative attack and the degradation is accompanied by fluorescence and decreasing absorbance. From enzymes engaged in the biotransformation of polycyclic hydrocarbons only phagosomal NADPH oxidase meets the criteria (particularly as for compartmental and catalytic properties) to be involved in melanin biodegradation. The in vivo biodegradation of melanin has so far been clearly demonstrated in Aspergillus and fungi melanins. PMID- 12753403 TI - The role of Kit-ligand in melanocyte development and epidermal homeostasis. AB - Kit-ligand (Kitl) also known as steel factor, stem cell factor and mast cell growth factor plays a crucial role in the development and maintenance of the melanocyte lineage in adult skin. Kitl exerts permanent survival, proliferation and migration functions in Kit receptor-expressing melanocytes. A comprehensive overview over the differential roles of Kitl in melanocyte development and homeostasis is provided. I discuss species-specific differences of the Kitl/Kit signalling system, regulation at the transcriptional level and also covering the regulation of cell surface Kitl presentation by cytoplasmic targeting sequences. In addition, recent studies evoked the importance of Kitl misexpression in some hyperpigmented lesions that may open the avenue for Kitl-dependent treatment of pathological skin conditions. PMID- 12753404 TI - Acquired melanocytic nevi as risk factor for melanoma development. A comprehensive review of epidemiological data. AB - Acquired melanocytic nevi (MN) in Caucasian populations are important markers for the risk of melanoma development. The total number of MN on the whole body is the most important independent risk factor for melanoma and the risk of melanoma development increases almost linearly with rising numbers of MN. Additionally, the presence of atypical MN and of actinic lentigines are likewise independent risk factors for melanoma. Atypical mole syndrome should be defined by the presence of many acquired MN and a threshold number of atypical MN. Acquired MN develops mainly during childhood and adolescence in the first two decades of life. The number of acquired nevi seems to be related to hereditary factors and nevus-prone families exist. The amount of sun exposure is the most important environmental risk factor for nevus development, particularly in early childhood. Interestingly, sunburns may play a role in nevus development, but seem not to be required, and even moderate sun exposure promotes the process. Therefore, preventive measures for nevus and melanoma development should target young children and adolescents. PMID- 12753405 TI - Oculocutaneous albinism type 1: the last 100 years. AB - Research on human albinism has been central to many of the major discoveries in human genetics. These include the first evidence that Mendel's rules of genetic segregation apply to humans, first published in 1903. Contrary to initial thought that albinism is caused by mutations in a single gene, we now know that the genetics of albinism are complex. The complexity of albinism was hinted at, in early publications, but has only recently been fully appreciated with the advent of molecular techniques. Currently, 12 different genes have been identified, that when mutated, result in a different type of albinism. Oculocutaneous albinism type 1 (OCA1), resulting from mutations of the tyrosinase gene, is genetically and biochemically the best understood type of albinism. Though much of the research in albinism has involved OCA1, there are many unanswered questions about OCA1 and albinism, in general. The next 100 yr should still provide many surprises as did the first 100 yr. PMID- 12753406 TI - Beta-catenin in the melanocyte lineage. AB - beta-Catenin is a multifunctional protein involved in cell-cell adhesion, intracellular signalling and gene transcription. It has been implicated in the development of various lineages, including neural crest derivatives. Melanocytes are derived from neural crest cells and beta-catenin is expressed throughout the development of this cell lineage. The multifunctional activity of beta-catenin is directly associated with its participation in multi protein-protein interactions. The cell-cell adhesion function of beta-catenin is mediated by the large cadherin cell adhesion molecule family, the intracellular signalling function by its interaction with GSK3beta, and the gene transcription activity by the four known LEF/TCF DNA binding-proteins. Here, we review the known beta-catenin interacting factors and targets involved in the development and transformation of melanocytes and in particular its role in the expression of the crucial gene of melanocyte development, Mitf. PMID- 12753408 TI - Regulation of 1-alpha, 25-dihydroxyvitamin D3 on interleukin-6 and interleukin-8 induced by sulfur mustard (HD) on human skin cells. AB - The regulatory effects of the active form of vitamin D, 1-alpha, 25 dihydroxyvitamin D3 (1-alpha, 25 (OH)2D3) were assessed on the cytokine and chemokine secretion induced by sulfur mustard on human skin fibroblasts and human epidermal keratinocytes. Stimulation of human skin fibroblasts with sulfur mustard (10(-4) M for 24 hr at 37 degrees ) resulted in approximately a 5 times increase in the secretion of interleukin-6 and over a 10 times increase for interleukin-8, which was inhibited by 1-alpha, 25 (OH)2D3, at 100 kDa by membrane filtration, is heat labile and sensitive to either protease digestion or reduction with DTT. Immunoblotting showed that mucus and supernatants of ultracentrifuged mucus from immune sheep contained IgG1 and IgA antibodies that recognized predominantly a larval antigen with an estimated molecular weight of 35 kDa on SDS-PAGE. Antibodies eluted from the surface of washed larvae that had been incubated in immune mucus also reacted specifically with the 35 kDa antigen on blots of larval homogenate. Immunofluorescence and immunogold electron microscopy showed that the 35 kDa antigen is present on the epicuticle of L3 and is shed during the moult to L4. The antigen is not present in eggs, L1, L2, L4 or adult worms and is found only in extracts of sheaths and L3 before infection and up to 4 days after infection. We hypothesize that the binding of antibody to the larval surface prevents larvae from establishing at their preferred site, causing them to be eliminated from the intestine. Monoclonal antibody PAB-1 recognizes the 35 kDa T. colubriformis larval antigen and also cross-reacts with antigens of similar molecular weight on blots of L3 extracts of the parasitic nematodes Haemonchus contortus and Ostertagia circumcincta; and with a 22-kDa antigen on blots of L3 extracts from Cooperia curticei and Nematodirus spathiger. This indicates that an antigenically related surface antigen with immunizing potential is present on several nematode species and can be identified by mAb PAB-1. The 35 kDa T. colubriformis larval antigen and related molecules in other nematodes are potential novel targets for stimulating host-protective immunity against nematode infections. PMID- 12753438 TI - Drug development for children: the past, the present, hope for the future. PMID- 12753440 TI - Brain injury in children with congenital heart disease. AB - The incidence of neurodevelopmental impairment in children with congenital heart disease is high. Its aetiology is multiple and complex. Prevention and treatment must start during the preoperative period and continue through the intra- and postoperative periods. Research has resulted in a clearer understanding of the relationship between congenital heart disease and the brain, and of the effects of cardiopulmonary bypass, hypothermia and circulatory arrest. This has led to modifications in management which may improve neurological outcome in the future. PMID- 12753441 TI - The dynostatic algorithm accurately calculates alveolar pressure on-line during ventilator treatment in children. AB - BACKGROUND: Monitoring of respiratory mechanics during ventilator treatment in paediatric intensive care is currently based on pressure and flow measurements in the ventilator or at the Y-piece. The characteristics of the tracheal tube will modify the pressures affecting the airways and alveoli in an unpredictable manner. The dynostatic algorithm (DSA), based on a one-compartment lung model, calculates the alveolar pressure during on-going ventilation. The DSA is based on accurate measurement of tracheal pressure. The purpose of this study was to test the validity of the DSA in a paediatric lung model and to apply the concept in an observational clinical study in children. METHODS: We validated the DSA in a paediatric lung model with linear, nonlinear pressure flow and frequency dependent characteristics by comparing calculated dynostatic (alveolar) pressures with directly measured alveolar pressures in the model and proximal plateau pressure with maximum alveolar pressure. Sixty combinations of ventilation modes, positive end expiratory pressures, inspiratory : expiratory ratios, volumes and frequencies were studied. A 0.25-mm fibreoptic pressure transducer in the tube lumen was used in combination with volume and flow from ventilator signals. Clinical measurements were performed in eight patients during anaesthesia and postoperative ventilator treatment. RESULTS: In the lung model we found a correlation coefficient between calculated and measured alveolar pressure of 0.93 0.99 with root mean square median values of 1 cm H2O. Distal plateau pressure agreed well with maximum alveolar pressure. In the clinical situation, the algorithm provided a breath-by-breath display of the volume-dependent lung compliance and the temporal course of alveolar pressure during uninterrupted ventilation. CONCLUSIONS: Fibreoptic measurement of tracheal pressure in combination with the dynostatic calculation of alveolar pressure provides an on line monitoring of the effects of ventilatory mode in terms of volume-dependent compliance, tracheal peak pressure and true positive end expiratory pressure. PMID- 12753442 TI - Increased respiratory symptoms following surgery in children exposed to environmental tobacco smoke. AB - OBJECTIVE: The aim of this study was to determine if children exposed to environmental tobacco smoke (ETS) via parental smoking (ETS+) developed more respiratory symptoms resulting in longer recovery times following surgical outpatient procedures compared with children of nonsmoking parents (ETS-). METHODS: One hundred and forty six children (4.9 +/- 3 years) undergoing inguinal hernia repair were prospectively studied. Parental smoking behaviour was determined by survey and urine analysis. Seven respiratory symptoms were evaluated during induction and emergence from anaesthesia and during the recovery room (RR) stay. RESULTS: Fifty-seven (39%) families admitted a smoking history while 89 (61%) denied it. This strongly correlated with the cotinine/creatinine ratio (Pearson correlation coefficient = 0.76; P = 0.01). ETS exposure was associated with an increased frequency of RR symptoms (ETS+: 56%; ETS-: 31%; P = 0.007). CONCLUSIONS: In children undergoing general anaesthesia for inguinal hernia repair, ETS exposure was associated with an increased frequency of respiratory symptoms during emergence from anaesthesia and during postoperative recovery. PMID- 12753443 TI - A randomized trial of caudal block with bupivacaine 4 mg x kg-1 (1.8 ml x kg-1) plus morphine (150 microg x kg-1) vs general anaesthesia with fentanyl for cardiac surgery. AB - BACKGROUND: Regional anaesthesia has been used effectively in paediatric patients undergoing cardiac surgery and is thought to be safe. METHODS: Thirty patients ASA physical status II-III undergoing scheduled palliative or corrective cardiac surgery, receiving premedication with midazolam and anaesthetic induction with sevoflurane, fentanyl and pancuronium were randomly allocated to two groups. In group 1, patients received bupivacaine 0.22% 4 mg.kg-1 (1.8 ml.kg-1) and morphine 150 microg x kg-1 by the caudal route. After a 20-min period for the block to take effect, sevoflurane 0.5-1.0% and fentanyl 5 microg x kg-1 were administered for maintenance of anaesthesia. In group 2, the anaesthetic technique was the same as in group 1, without a caudal block and fentanyl 25 microg x kg-1 was administered at the moment of surgical incision. RESULTS: Cardiovascular and haemodynamic responses of patients receiving caudal block showed minor variations during the 20-min period between caudal and general anaesthesia. Fentanyl requirements during surgery were lower (P = 0.001) in patients with caudal block than patients with general anaesthesia. Extubation time was shorter (P = 0.034) in the caudal group. Two patients in the general anaesthesia group and one in the caudal group died because of postoperative complications. CONCLUSIONS: Caudal block with bupivacaine 0.22% 4 mg.kg-1 (1.8 ml.kg-1) and morphine 150 microg x kg 1 was safe and effective for paediatric patients undergoing cardiac surgery. However, patients might have a better outcome with a reduction of morphine dosage and administration of a muscle relaxant of shorter duration of action than pancuronium. PMID- 12753444 TI - Side-effects of postoperative epidural analgesia in children: a randomized study comparing morphine and clonidine. AB - BACKGROUND: Morphine is widely used in association with local anaesthetics for postoperative epidural analgesia. There are no data on the prolonged use of clonidine for postoperative analgesia in children. The primary outcome of this randomized, double-blind trial was to compare the incidence of side-effects after epidural infusion of clonidine or morphine, in association with ropivacaine in children. METHODS: After institutional approval, 26 children, aged 3-12 years, who were scheduled for abdominal surgery, had an epidural catheter placed after induction of general anaesthesia. Patients were then randomized to two different groups. After an initial bolus of 2.5 mg x kg-1 0.25% ropivacaine with either 40 micro g x kg-1 morphine (group M, n = 14) or 1 micro g x kg-1 clonidine (group C, n = 12), an epidural infusion was started at a rate of 0.4 ml x kg-1 x h-1. The patients in the M group received an infusion of 0.08% ropivacaine with 10 micro g.ml-1 morphine, those in the group C an infusion of 0.08% ropivacaine with 0.6 micro g.ml-1 clonidine. RESULTS: The two groups were similar with respect to age, sex and weight. One patient in the C group was excluded for misplacement of the epidural catheter. The incidence of vomiting and pruritus was significantly higher in the M group compared with the C group (64% and 85% versus 0%, respectively). The incidence of pain was significantly higher in the C group compared with the M group (73% versus 29%) as well as the need for rescue analgesia medications. CONCLUSIONS: Epidural clonidine is followed by a significantly lower incidence of side-effects. However, its analgesic effects, at least at the doses used in this study, are less potent than those of epidural morphine. PMID- 12753446 TI - Dorsal penile nerve block. AB - BACKGROUND: The dorsal penile nerve block (DPNB) is a common form of regional anaesthesia for circumcisions in children. The safety of this block has been questioned following several case reports of potentially serious complications. METHODS: We performed a retrospective review of all the incidents related to the conduct of this block in our institution over a period of 53 months. RESULTS: A total of 3909 circumcisions were performed under DPNB and nine incidents were recorded. This included two cases of bleeding from the prepuce, two urethral injuries, three haematomas and two drug errors. Two cases required further surgical intervention but all of the patients recovered with no apparent long term sequelae. CONCLUSIONS: We feel that the DPNB is a safe technique for circumcisions in infants and children. PMID- 12753445 TI - Caudal neostigmine for postoperative analgesia in paediatric surgery. AB - BACKGROUND: This study was conducted to evaluate analgesia and side-effects of caudal neostigmine coadministered with bupivacaine in paediatric surgery. METHODS: We studied children, aged 1-5 years, undergoing elective surgery (inguinal hernia and hypospadias). After standard induction of anaesthesia, caudal anaesthesia was performed. Group 1 received 0.25% bupivacaine 0.5 ml.kg-1 and Group 2 received 0.25% bupivacaine 0.5 ml x kg-1 with 1 microg x kg-1 neostigmine via the caudal route. Heart rate, mean arterial pressure, peripheral oxygen saturation were recorded before induction, after induction but before caudal anaesthesia, and then every 5 min after caudal anaesthesia. Haemodynamic, Toddler, Preschooler, Postoperative Pain Scale (TPPPS) pain score and sedation score values were recorded 30 min after extubation and at hours 2, 4, 6, 12 and 24. A pain score >3/10 resulted in administration of rectal paracetamol. The duration of postoperative analgesia was defined as the time between caudal drug injection and the first rectal paracetamol administration. RESULTS: There were no differences between the groups in demographic and haemodynamic date, duration of surgery and anaesthesia, time to extubation or sedation scores. The duration of postoperative pain relief did not differ between the two groups; 15.40 +/- 10.97 h for group 1 vs. 15.45 +/- 10.99 h for group 2 (P > 0.05). The incidence of nausea (three patients in group 2 and one patient in group 1) was not statistically significant. No other side-effects were seen. CONCLUSIONS: We found that a single caudal injection of 1 microg x kg-1 neostigmine mixed with bupivacaine offers no significant advantage over bupivacaine alone for postoperative pain relief in children undergoing genitourinary surgery. PMID- 12753448 TI - Life threatening subcutaneous emphysema following surgical repair of tracheocutaneous fistula. AB - A 9-year-old male child posted for closure of tracheocutaneous fistula developed extensive subcutaneous emphysema resulting in acute respiratory distress immediately after transfer to recovery room. The clinical management, precautions and other complications of closure of tracheocutaneous fistula are discussed. PMID- 12753447 TI - Increased incidence of postoperative nausea and vomiting without additional analgesic effects when a low dose of intravenous fentanyl is combined with a caudal block. AB - BACKGROUND: The use of opioids is known to increase the incidence of postoperative nausea and vomiting (PONV). In spite of this, administration of low doses of an opioid during anaesthesia is common practice, even if a regional anaesthetic technique is used. This study was designed to estimate the effects of intraoperative intravenous administration of fentanyl on PONV in paediatric daycase surgery. METHODS: PONV and pain were evaluated in 29 boys during the first 24 h after daycase penile surgery. Anaesthesia was standardized. The patients were allocated to two groups. Fentanyl 1 micro g.kg-1 i.v. or placebo was administered in a randomized, double-blind design. A caudal block with ropivacaine 2 mg.ml-1, 0.5 ml.kg-1 was performed preoperatively and topical lidocaine gel 20 mg.ml-1 was applied over the wound area immediately after surgery. RESULTS: The total incidence of PONV in hospital and at home during the first 24 h was 36% (5/14) when fentanyl was used, whereas no PONV was reported when placebo was given (P < 0.05). The median time to first administration of analgesics after the caudal block was approximately 6 h. It did not differ between groups. Intraoperative fentanyl did not result in any reduction in pain scores nor the incidence of pain. Fentanyl caused apnoea in one-half of the cases and decreased the breathing rates during the first 10 min of surgery. CONCLUSIONS: Intraoperative use of i.v. fentanyl 1 micro g.kg-1 combined with a regional anaesthetic block is associated with an increased incidence of PONV without any significant contribution to the postoperative pain relief. PMID- 12753449 TI - Difficult airway in a child with spinal muscular atrophy type I. AB - Spinal muscular atrophy (SMA) type I is a relatively common inherited neuromuscular disease of hypotonic newborns, but is not associated with craniofacial abnormalities. There is nothing in the literature about difficult intubation in patients affected by this disease. We report a case of 34-month-old girl with SMA type I who was scheduled for emergency endoscopic laser treatment of tracheal stenosis caused by granulations. Tracheostomy was performed at 17 months of age and before this, the orotracheal tube was changed periodically without difficulty. For this laser treatment, orotracheal intubation was required. Preoperative physical examination revealed micrognathia and class II malocclusion. Opening her mouth was not difficult. Although difficult orotracheal intubation was predictable, we attempted to intubate her trachea as usual, but could not visualize the epiglottis. We decided to proceed with retrograde intubation, one of the standard techniques employed in a child with a difficult airway, via the tracheostome. A feeding nasogastric catheter was used as a guide catheter, and our strategy was successful. In this study we report a case of difficult airway in a child with SMA type I. The relationship between SMA type I with a tracheostome and difficult airway are discussed. PMID- 12753450 TI - Complete staged palliation of hypoplastic left heart syndrome in a child with cystic fibrosis. AB - In recent years, the palliative treatment of Hypoplastic Left Heart Syndrome (HLHS) with a three-staged surgical repair has gained widespread acceptance in North America and elsewhere and has significantly improved the life-expectancy of these children. We report on a child in whom the diagnosis of cystic fibrosis (CF) was made shortly after the first-stage palliation (modified Norwood procedure), and in which surgical palliation was successfully completed with second- (bidirectional Glenn) and third-stage (fenestrated Fontan) procedures. During this period, the child suffered several CF-related complications, i.e. repeated respiratory failure, meconium ileus and coagulation abnormalities, which required adjustment of the anaesthesia, surgical and intensive care management. Considering the trend to perform surgical corrections of major cardiac defects in the early neonatal period, before concomitant genetic diseases are discovered, paediatric cardiac anaesthesiologists, surgeons and intensivists may be confronted with more cases of open heart surgery in infants with CF in the future. PMID- 12753451 TI - Flumazenil is innocuous in a paroxysmal supraventricular tachycardia and radiofrequency ablation: a paediatric case report and review of the literature. AB - The efficacy and safety of flumazenil in reversing midazolam-induced paradoxical reaction in a child suffering from paroxysmal supraventricular tachycardia (PSVT) and undergoing radiofrequency ablation procedure has not been described before. We report a boy who had suffered for years from poorly controlled PSVT because of noncompliance to medications and was being sedated by midazolam and morphine (4 mg each) and prepared for radiofrequency ablation when he began attempting to speak, writhing and flailing his arms, for which physical restraint was required. We also review the current knowledge on the subject. Two doses of flumazenil 0.05 mg i.v. swiftly halted the untoward episode. The child opened his eyes and was calm, coherent and able to answer simple questions. All haemodynamic and respiratory parameters remained stable throughout this event, except for a temporary increase in heart rate (from 68 to 83 b x min-1). The aberrant behaviour did not resume when he was allowed to fall asleep during the ablation of a concealed left lateral accessory pathway. After that procedure, he awoke calm and pain free with no recollection of the aggressive episode. He was kept for 24 h in an intermediate cardiac care unit and was then discharged home. This apparent first report on the efficacious use of flumazenil for the reversal of a paradoxical reaction to midazolam during an electrophysiological study suggests that it neither induces abnormal heart rate nor interferes with the electrophysiological study or the ablation procedure. PMID- 12753452 TI - Bispectral index in a 3-year old undergoing deep hypothermia and circulatory arrest. AB - We report a 3-year-old girl who presented with Scimitar syndrome and underwent hypothermic circulatory arrest for correction of anomalous pulmonary veins and an atrial septal defect. In this case the Bispectral Index (BIS) correlated significantly with the gradual onset of hypothermia and circulatory arrest. However, BIS remained low during the rewarming phase of cardiopulmonary bypass, in spite of adequate pump flows and stable haemodynamics. We postulate that this significant lag in BIS during the rewarming phase of deep hypothermic circulatory arrest may represent neuronal bewilderment or perhaps stunning, and differs from previous studies that show significant increase in BIS during rewarming from mild hypothermia. PMID- 12753453 TI - A combined stage 1 and 2 repair for hypoplastic left heart syndrome: anaesthetic considerations. AB - Therapy of hypoplastic left heart syndrome (HLHS) consists of the staged Norwood procedure or cardiac transplantation. Stenting the ductus arteriosus and subsequent banding of the pulmonary arteries allows the combination of neoaortic reconstruction with the establishment of a bidirectional cavopulmonary connection (combined stage 1 and 2 procedure) in a later session. We report the anaesthetic management in eight infants ranging from 107 to 195 days undergoing a combined stage 1 and 2 procedure. Nonselective pulmonary vasodilators and nitric oxide were needed in all cases to improve oxygen saturation in the postbypass period. Phosphodiesterase inhibitors and epinephrine were required in all patients for inotropic support during and after weaning off cardiopulmonary bypass. The procedure was successful in seven patients. One patient died intraoperatively because of right heart failure. The physiological changes of this new surgical strategy for palliation of HLHS offers a challenge for the anaesthetist primarily in the early postbypass period. PMID- 12753454 TI - Response to Martay et al. PMID- 12753455 TI - Instrumentation of the airway in the absence of intravenous access. PMID- 12753456 TI - Urethral meatal dilatation using topical EMLA cream for anaesthesia. PMID- 12753457 TI - Postdural puncture headache in children. PMID- 12753459 TI - Flumazenil reverses a paradoxical reaction to intravenous midazolam in a child with uneventful prior exposure to midazolam. PMID- 12753460 TI - Response to review by Herod et al. PMID- 12753462 TI - Distribution of Actinobacillus actinomycetemcomitans serotypes and Porphyromonas gingivalis in Japanese adults. AB - Strains of the bacterium Actinobacillus actinomycetemcomitans found in the human oral cavity are divided into five serotypes, a, b, c, d, and e. In this study, A. actinomycetemcomitans serotypes and Porphyromonas gingivalis were isolated from 656 subgingival sites in systemically healthy Japanese adults. A. actinomycetemcomitans was detected in 19.5% of 328 Japanese subjects, while 27.1% of subjects were positive for P. gingivalis. Of 75 A. actinomycetemcomitans positive sites, only one serotype was detected in 39 sites (52.0%). The numbers of sites in which two different serotypes and three different serotypes were detected were 18 (25.0%) and 7 (9.3%), respectively. A. actinomycetemcomitans serotype c was detected more frequently in sites that were positive for both P. gingivalis and A. actinomycetemcomitans (76.9%) than in sites that were P. gingivalis-negative and A. actinomycetemcomitans-positive (33.9%). In addition, serotype c was detected much more frequently than the other serotypes (<16%) in sites that were positive for both P. gingivalis and A. actinomycetemcomitans. These findings suggest that the characteristics of serotype c may differ from those of the other serotypes. This report is the first to use PCR to describe the distribution of A. actinomycetemcomitans serotypes in humans and to examine the association between the distribution of A. actinomycetemcomitans serotypes and the presence of P. gingivalis. PMID- 12753463 TI - Effectiveness of dentine bonding agents against cariogenic bacteria in vitro: a comparison of two methods. AB - Data obtained from studies on the antimicrobial properties of bonding agents are the subject of controversy, probably because of methodological differences. This study compared two commonly used in vitro methods, the disc agar diffusion test and the well agar diffusion test. Agar plates were seeded with Streptococcus sobrinus, Lactobacillus gasseri, or Actinomyces naeslundii. For the well diffusion test, wells cut out of the agar were filled with the test material, and for the disc method, discs impregnated with the test material were applied to the agar; the discs and wells were both 9 mm in diameter. After incubation, measurements of the zones of inhibition showed little agreement between the two methods when bonding agents were tested; the mean differences (+/- sdiff) in the zones of inhibition between the methods were 0.7 +/- 3.4 mm (P = 0.40, one sample t-test against zero), 4.9 +/- 4.4 mm (P = 0.97), and 0.8 +/- 4.3 mm (P = 0.47) for S. sobrinus, L. gasseri, and A. naeslundii, respectively. Mean differences were less contrasting when chlorhexidine and pure components were tested (P < 0.05 for S. sobrinus and L. gasseri). These results indicate the need for a gold standard method to evaluate the antimicrobial properties of bonding agents. PMID- 12753464 TI - Determination of mutacin activity and detection of mutA genes in Streptococcus mutans genotypes from caries-free and caries-active children. AB - Relationships between genetic diversity, mutacin production and sensitivity to mutacins in Streptococcus mutans were evaluated in 19 clinical isolates from caries-free and caries-active children. Mutacin production was tested against 30 indicator strains; results showed significant variations in the inhibitory spectra of the clinical isolates. There was no association between the inhibitory spectrum of the infecting strain and the caries experience or the level of mutans streptococci infection of the host. Homology to the mutA gene coding for mutacin II was detected in one clinical isolate; none of the clinical isolates showed homology to the mutA genes coding for mutacins I or III. Genotyping by random amplified polymorphic DNA (RAPD) reactions grouped the isolates into three clusters, but no correlation was found between any of the clusters and mutacin activity, caries experience or level of mutans streptococci in the host. PMID- 12753465 TI - Prostaglandin E2 downregulates interleukin-12 production through EP4 receptors in human monocytes stimulated with lipopolysaccharide from Actinobacillus actinomycetemcomitans and interferon-gamma. AB - In the present study, we examined the effect of prostaglandin (PG) E2 on interleukin (IL) -12 production in monocytes stimulated with a combination of lipopolysaccharide (LPS) from Actinobacillus actinomycetemcomitans and interferon gamma (A. actinomycetemcomitans-LPS/IFN-gamma). Indomethacin, a cyclooxygenase inhibitor, enhanced IL-12 production, but inhibited PGE2 generation in A. actinomycetemcomitans-LPS/IFN-gamma-stimulated monocytes. Exogenous PGE2 inhibited IL-12 release in the cells. EP2, EP3 and EP4 receptor mRNA expression was detected in monocytes by reverse transcription-polymerase chain reaction. 11 deoxy-PGE1 (an EP2/EP4 agonist) inhibited IL-12 production in A. actinomycetemcomitans-LPS/IFN-gamma-challenged monocytes, whereas butaprost (an EP2 agonist) or ONO-AP-324 (an EP3 agonist) had no effect on IL-12 production. Dibutyryl cAMP, a cAMP analogue, and forskolin, an adenylate cyclase activator, mimicked depression of IL-12 production by PGE2. From these results, we suggest that PGE2 inhibits IL-12 production via EP4 receptors by cAMP-dependent pathways in A. actinomycetemcomitans-LPS/IFN-gamma-challenged monocytes. PMID- 12753466 TI - PCR method is essential for detecting Mycobacterium tuberculosis in oral cavity samples. AB - Tuberculosis is a re-emerging infectious disease, and infection by Mycobacterium tuberculosis has been increasing in immunocompromised hosts, including elderly persons. M. tuberculosis-infected persons may receive dental treatment. To evaluate the risk of M. tuberculosis infection in dental clinics, we examined the detection rates of M. tuberculosis in sample of mixed saliva, dental plaque, extracted teeth, caries lesions, and denture plaque by nested polymerase chain reaction (PCR). The detection rates by PCR in samples from mixed saliva, dental plaque, caries lesions and denture plaque obtained from tuberculosis patients were 98.0%, 92.0%, 89.0%, and 100%, respectively. The detection rates by the culture method were 17.3%, 2.0%, 0%, and 0%, respectively. M. tuberculosis also was detected from the nontuberculous mycobacteria-infected group. Strains of Actinomyces naeslundii, Porphyromonas gingivalis, and Fusobacterium nucleatum inhibited the growth of clinical strains of M. tuberculosis, but strains of Streptococcus mutans, Streptococcus sanguinis, and Actinobacillus actinomycetemcomitans did not. The present study concludes that the PCR method is essential for detecting M. tuberculosis in oral samples. PMID- 12753467 TI - Mutans streptococci strains prevalence before and after cavity preparation during Atraumatic Restorative Treatment. AB - Critics argue that all carious dentine is not removed from the hand-prepared cavity during the Atraumatic Restorative Treatment (ART) procedure, and that the caries process is soon resumed. The aim of this study was to determine the effectiveness of ART in removing carious tissue, by investigating the numbers of mutans streptococci and lactobacilli, with emphasis on the prevalence of Streptococcus mutans and Streptococcus sobrinus strains before, and after ART treatment of dental caries. Two microbiology samples were collected. The first sample was removed from the centre of the carious lesion at the enamel-dentine junction, and the second was collected from the centre of the hard cavity wall above the pulp, after the soft infected dentine had been manually removed. A total of 71 mutans streptococci isolates from 31 children and 40 carious teeth were subcultured, biochemically characterised and genotyped by the arbitrarily primed polymerase chain reaction (AP-PCR). Results showed a significant decrease in TVC (P<0.0001), mutans streptococci (P < 0.0001) and lactobacilli (P = 0.0002) after cavity preparation. AP-PCR identified S. mutans strains that were undetectable during biotyping, and divided clinical isolates into two main clusters. In all, 63% (45/71) of isolates from the carious lesions comprised S. mutans strains. After cavity preparation, this was reduced to 35% (25/71), of which 30% (21/71) were S. mutans and the remaining 6% (4/71) S. sobrinus strains. The number of mutans streptococci strains was below detectable levels in 19 of the prepared cavities. The significant decrease in bacteria after manual cavity preparation demonstrates the reliability of a standardized ART technique, yet the presence of S. mutans strains shows that the effectiveness of the ART procedure can vary during treatment and between dental practitioners. PMID- 12753468 TI - Granulocyte-macrophage colony-stimulating factor responses of oral epithelial cells to Candida albicans. AB - Candida albicans is the principal fungal species responsible for oropharyngeal candidiasis, the most frequent opportunistic infection associated with immune deficiencies. Cytokines, such as granulocyte-macrophage colony-stimulating factor (GM-CSF), are important in the generation of effective immunity to C. albicans. The purposes of this investigation were to determine whether C. albicans triggers secretion of GM-CSF by oral epithelial cells in vitro and to investigate mechanisms of host cell-fungal interactions that trigger such responses. Oral epithelial cell lines as well as primary oral mucosal epithelial cells were challenged with stationary phase viable C. albicans, added to human cell cultures at varying yeast:oral cell ratios. Yeast were allowed to germinate for up to 48 h and supernatants were analyzed for GM-CSF by ELISA. Fixed organisms, germination deficient mutants and separation of yeast from epithelial cells using cell culture inserts were used to assess the effects of viability, germination and physical contact, respectively, on the GM-CSF responses of these cells. Two out of three cell lines and three out of six primary cultures responded to C. albicans with an increase in GM-CSF secretion. GM-CSF responses were contact dependent, strain-dependent, required yeast viability and were optimal when the yeast germinated into hyphae. PMID- 12753469 TI - Classification of oral streptococci by two-dimensional gel electrophoresis with direct activity stain for glycosyltransferases. AB - Thirty eight strains of oral streptococci were divided into six types by two dimensional gel electrophoresis (2-DE) followed by glycosyltransferase (GLT) activity stain: Type 1, Streptococcus mutans; Type 2, Streptococcus rattus; Type 3, Streptococcus sobrinus and Streptococcus downei; Type 4, Streptococcus cricetus; Type 5, Streptococcus salivarius; and Type 6, Streptococcus sanguis, Streptococcus oralis and Streptococcus gordonii. In Types 1, 2 and 5, two glucosyltransferases synthesizing water-insoluble (GTF-I) and water-soluble glucans (GTF-S) and a fructosyltransferase (FTF) were detected. In Types 3 and 4, GTF-I and two GTF-Ss were detected. Only one GTF-S was detected in Type 6. The 2 DE patterns for these six types were clearly distinguished from each other based on the kind, number and location of GLTs in gel. PMID- 12753470 TI - The IgG antibody profile to various antigen regions of hepatitis C virus differs in oral fluid and serum of patients with chronic hepatitis C. AB - Antibodies to hepatitis C virus (HCV) can be detected not only in serum but also in oral fluid. The aim of the study was to determine IgG antibody reactivity directed to six antigen regions of HCV in oral fluid and to evaluate the significance of the antibody pattern in oral fluid compared to serum. Oral fluid and serum samples of 32 HCV viremic patients were collected to detect antibodies to six antigen regions incorporated as antigen bands into modified commercial updated third generation line immuno-assay. Compared to serum, a significantly lower cumulative antibody response and reactivity to five HCV antigens was found in oral fluid. The significantly highest prevalence of oral fluid reactivity was recorded with antigen C1 (78%), whereas in serum the most significantly frequent reactivity was detected with antigen NS3 (100%). The absence of antibody reactivity with antigen E2 was similar in both body fluids. The discrepancy in antibody pattern to HCV antigens between oral fluid and serum indicates the possible existence of local viral replication, viral mutants, viral inhibitors in oral cavity and, most probably, leakage of the muco-vascular barrier. PMID- 12753472 TI - Inhibitory effects of green tea catechins on protein tyrosine phosphatase in Prevotella intermedia. AB - Members of the Prevotella intermedia group possess protein tyrosine phosphatase (PTPase). The purpose of this study was to investigate the effects of catechin derivatives from Japanese green tea on the activity of PTPase in P. intermedia and related organisms. Multilocus enzyme electrophoresis of alkaline phosphatase derived from P. intermedia, Prevotella nigrescens, Prevotella pallens and Porphyromonas gingivalis revealed a species-specific migration pattern. Among the tea catechin derivatives tested, (-)-epigallocatechin gallate (EGCg), similar to orthovanadate, a specific inhibitor for PTPase, was effective in inhibiting the PTPase activity in P. intermedia at 0.5 microm, and related species at 5 microm. The results suggested that the inhibitory effect observed is due to the presence of galloyl moiety in the structure. In contrast, neither the green tea catechins nor orthovanadate inhibited the phosphatase activity in P. gingivalis, suggesting that this organism possessed a different family of alkaline phosphatase. PMID- 12753471 TI - Characterization of the immunologic responses to human in vivo acquired enamel pellicle as a novel means to investigate its composition. AB - Human acquired enamel pellicle is formed by molecules selectively adsorbed onto tooth surfaces. The present work describes the use of monoclonal antibody (mAb) technology as a novel approach to identify micro amounts of components present in pellicle. MAbs were obtained with reactivities against statherin, histatin, mucous glycoprotein 1(MGI), albumin, amylase and human immunoglobulins (Igs), indicating that these are pellicle components, which was further confirmed by immunoblotting. No mAbs against proline-rich proteins (PRPs), lysozyme, mucous glycoprotein 2 (MG2), carbonic anhydrase, lactoferrin or peroxidase were obtained, suggesting that these components are absent, present in low amounts, or exhibit low antigenicity. Further characterization of the binding epitopes of some of th e obtained anti-MGO, anti-statherin and anti-histatin mAbs were carried out and the biological relevance is discussed. The results open up the possibility that immunization with human pellicle and mAbs production can be employed to identify hitherto unknown constituents of pellicle. PMID- 12753473 TI - Purification and partial characterization of a dipeptidyl peptidase from Prevotella intermedia. AB - A peptidase hydrolyzed X-Pro-p-nitroanilide was purified from the cell extract of Prevotella intermedia ATCC 25611 by ion-exchange chromatography and hydrophobic interaction chromatography. The purified enzyme exhibited a molecular size of 74 kDa from sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the maximum enzyme activity was found between pH 7.0 and pH 7.5. This peptidase was a serine enzyme and hydrolyzed Lys-Pro-p-nitroanilide, Arg-Pro-p-nitroanilide, and Ala-Pro-p-nitroanilide, but Lys-Ala-p-nitroanilide was not split. The enzyme may be classified as a dipeptidyl peptidase IV. PMID- 12753474 TI - Periodontal pathogens in subgingival plaque of HIV-positive subjects with chronic periodontitis. AB - Many putative periodontal pathogens associated with periodontal disease in human immunodeficiency virus (HIV)-infected patients also occur in non-HIV-infected individuals. This study examined the prevalence of eight periodontal pathogens in HIV-positive and HIV-negative patients with chronic periodontitis using the 16s RNA polymerase chain reaction technique. The results showed a significant prevalence of Porphyromonas gingivalis and Treponema denticola among HIV-negative patients compared to HIV-positive patients. Sixty percent of the patients in both groups were colonized by five to six species. Odds ratio analysis revealed a statistically significant positive association between three of the 28 possible combinations in the HIV-positive group. They included Prevotella nigrescens/Campylobacter rectus, P. nigrescens/P. gingivalis and P. nigrescens/T. denticola. Although the prevalence of periodontal pathogens is similar in both the groups, the combination of certain periodontal pathogens may be responsible for chronic periodontitis seen in HIV-infected adults. PMID- 12753475 TI - A corneal model for slit-scanning elevation topography. AB - Computerised videokeratography is increasing in sophistication and accuracy. The Orbscan II Videokeratographer combines the advantages of Placido derived and slit scanning derived topography, to acquire shape information on the posterior surface of the cornea. The purpose of this paper is to construct a model of slit scanning elevation topography and highlight potential sources of variation in this methodology. Ray tracing calculations were performed on a defined, theoretical, spherical model to obtain a pachymetric measurement. These calculations were tested by comparing the pachymetry measurement derived from the model, over a refractive index shift from 1.376 to 1.400 with the pachymetry measurement obtained by Orbscan II, of a human eye, with an identical alteration in refractive index. The two methods of measurement differed by only 0.157 microm, suggesting that the model is a good representation of slit-scanning elevation topography, and that refractive index variations have only a minimal effect on Orbscan II pachymetry measurement. PMID- 12753476 TI - The detection of small simulated field defects using multifocal VEPs. AB - INTRODUCTION: The multifocal visual-evoked potential (mfVEP) has been widely investigated in the study of diseases of the visual system. However, the sensitivity of the mfVEP in objective detection of field defects has not been determined. This study investigates the variation of the mfVEP responses whilst simulating field defects by using different sizes of mask on the stimulus pattern. METHODS: Simulated field defects of four different sizes (2, 3, 5, and 7 degrees) at two different eccentricities (10 and 16 degrees) were generated on a standard mfVEP dartboard stimulus using opaque masks. These masks were placed at the centre of each dartboard sector and the modified stimuli were used to elicit mfVEPs from 10 normal subjects. The response densities and latencies of N1, P1 of the mfVEP were compared, without and with small simulated field defects. RESULTS: The minimum size of simulated field defect causing significant response density reduction in P1 and N1 was 5 degrees at both retinal eccentricities. N1 showed similar reduction in response density at both retinal eccentricities, but P1 showed larger reduction at the 10-degree location than at the 16-degree location. There was no change in latencies with simulated field defect at either location. CONCLUSIONS: The mfVEP is only sensitive to a simulated field defect equal to or larger than 5 degrees in diameter, and mfVEP has greater sensitivity at 10-degree eccentricity than at 16-degree eccentricity. PMID- 12753477 TI - Computerized calculation scheme for bitoric eikonic intraocular lenses. AB - Despite full correction of the corneal astigmatism with toric intraocular lenses, the retinal image is distorted and the lateral image-object magnification is different in different meridians. The purpose of this study is to describe an iteration strategy for tracing an axial pencil of rays through the 'optical system eye' containing astigmatic refractive surfaces with their axes at random to calculate a thick bitoric lens implant which eliminates image distortion. The capabilities of this computing scheme are demonstrated with two clinical examples. We present a mathematically straightforward computer-based strategy for the calculation of thick bitoric eikonic lens implants. The iteration algorithm is initialized with a spherical front and a toric back surface and stepwise decreases the image distortion by adding cylinder lenses to the front lens surface corrected by the toric lens back surface. Total magnification can be modulated by varying the front-to-back surface power of the thick lens. PMID- 12753478 TI - Ocular and visual defects amongst people with intellectual disabilities participating in Special Olympics. AB - PURPOSE: To analyse data from vision screening of people with intellectual disabilities from the UK participating in Special Olympics during the National Summer Games in Cardiff 2001, in order to determine visual status and access to eye care. METHODS: Athletes were invited to take part in vision screening, according to Special Olympics protocols. RESULTS: Testability rates were high. Findings confirmed the high prevalence of eye and vision defects reported in other studies of people with intellectual disabilities. In particular, visual acuity was below normal for most athletes. Significant improvement to acuity could be achieved with spectacles although correction after full refraction did not always provide normal vision. Over 40% of athletes undergoing full refraction had below normal corrected acuity. CONCLUSIONS: Athletes with intellectual disabilities were no more likely to access eye care and have adequate spectacle correction than other people with intellectual disabilities. Optometrists and carers need to be aware of the high prevalence of defects and the importance of regular eye examinations in people with intellectual disabilities. PMID- 12753479 TI - Correlation analysis of visual field thresholds and scanning laser ophthalmoscopic optic nerve head measurements in glaucoma. AB - In a cross-sectional study of 50 consecutive primary open-angle glaucoma cases, full threshold 30-2 visual fields on Humphrey Visual Field Analyser were correlated to stereometric parameters obtained using a scanning laser ophthalmoscope (HRT II). The strength of linear correlation between the stereometric parameters and visual thresholds was used to generate a correspondence map between the optic disc and the visual field. Rim/disc area ratio and cup/disc area ratio correlated with the visual thresholds at the highest number of points in the visual fields. Thresholds at superior hemifield locations correlated best with the rim/disc area ratio and cup/disc area ratio in the inferonasal sector and the ones in the inferior hemifield correlated best with the rim/disc area ratio and cup/disc area ratio in the superotemporal sector. Optic disc parameters correlated better with inferior field locations and non-edge points than superior field locations and edge points. The data indicate that rim/disc area ratio and cup/disc area ratio measurements on HRT II have a good correlation with visual field damage seen on automated visual fields. PMID- 12753480 TI - The repeatability and validity of dynamic retinoscopy in assessing the accommodative response. AB - Dynamic retinoscopy (DR) is an objective technique that has been used successfully to examine accommodative function of non-communicative patients. However, there are few studies examining its repeatability and validity. This paper addresses the repeatability and validity of a form of DR (a modified Nott technique), by comparing results to the Shin-Nippon SRW-5000 Autorefractor whose validity and repeatability have previously been demonstrated. Accommodative responses of 41 subjects (aged 6-43 years) were assessed using DR and the Shin Nippon SRW-5000 Autorefractor. Three different target demands were used (10, 6 and 4 D). DR was shown to be repeatable for all target demands (paired t-test, p > 0.05). The results demonstrated good agreement between the autorefractor and DR. The results of the present study suggest that DR is a repeatable and valid objective technique for assessing accommodative function. PMID- 12753481 TI - Image size magnification and power and dilation factors for optical instruments in general. AB - Traditional treatments of spectacle magnification for distant objects consider only stigmatic spectacle lenses and they compare the retinal image size in a refractively fully compensated eye with the image size in the uncompensated eye. Spectacle magnification is expressed as a product of two factors, the power and shape factors of the lens. The power factor depends on the position of the entrance pupil of the eye. For an eye with an astigmatic cornea, however, the position of the entrance pupil is not well defined. Thus, the traditional approach to spectacle magnification does not generalize properly to allow for astigmatism. Within the constraints of linear optics and subject to the restriction that the eye's iris remains the aperture stop, this paper provides a complete, unified and exact treatment for optical instruments in general. It compares retinal image size in a generalized sense (including image shape and orientation) for any instrument in front of an eye with that of the eye alone irrespective of whether the instrument compensates or not. The approach does not make use of the concept of the entrance pupil at all and it allows for astigmatism and for non-alignment of refracting elements in the instrument and in the eye. The concept of spectacle magnification generalizes to the concept of instrument size magnification. Instrument size magnification can be expressed as the product of two matrix factors one of which can be interpreted as a power factor (as back-vertex power) and the other factor for which the name dilation factor is more appropriate in general. The general treatment is then applied to a number of special cases including afocal instruments, spectacle lenses (including obliquely crossing thick bitoric lenses), contact lenses, stigmatic systems and stigmatic eyes. In the case of spectacle lenses, the dilation factor reduces to the usual shape factor. PMID- 12753482 TI - Oculocardiac reflex caused by contact lenses. AB - PURPOSE: To report two cases of patients who fell unconscious because of the oculocardiac reflex when attempting to wear contact lenses. CASE REPORTS: Case 1: A 15-year-old healthy boy came to our clinic to be fitted with contact lenses. As soon as a hard contact lens was inserted forcibly, he became unconscious. Case 2: A 22-year-old man fell unconscious the instant that the eye was compressed with a hard contact lens. Neither patient had used glasses or contact lenses before. Their blood pressure was decreased while unconscious. They recovered consciousness after about 10 min, and nausea and vomiting settled subsequently. Contact lens insertion was tried again carefully after 1 week avoiding compression of the eyes and there were no problems. They are currently using contact lenses without any problems. CONCLUSION: Insertion of contact lenses may rarely provoke the oculocardiac reflex. PMID- 12753483 TI - Open- and closed-loop regressions compared in myopic and emmetropic subjects. AB - Post-near task accommodation regressions have been measured in a number of studies to investigate the association between myopia and nearwork. The slow nature of the regression found when measuring both open- and closed-loop regressions in some subjects has been hypothesised to be associated with myopia development and progression. However, it is not known whether the speed and characteristics of the regression for an individual is the same under open- and closed-loop conditions. In this study we measured post-near task accommodation regression functions under open- and closed-loop conditions for myopic and emmetropic subjects. We compared the responses of each subject under the two conditions and found that some subjects responded differently depending on the condition (i.e. a fast regression under one condition and a slow regression under the other condition). When compared as a group, the myopic subjects showed significantly more prolonged open-loop regressions than closed-loop regressions. In terms of the effect on the post-task accommodation, the myopes appear to be more affected by the measurement conditions than the emmetropes in this study. The results highlight the importance of the type of accommodation stimulus when investigating the accommodation response. PMID- 12753484 TI - Assessment of reproducibility of measures of intraocular pressure and central corneal thickness in young white adults over a 16-h time period. AB - PURPOSE: The aims of this study were to obtain clinical measures of intraocular pressure (IOP) and central corneal thickness (CCT) and during waking hours to further assess both the nature of diurnal changes, and the reproducibility of any possible association between the two measures. METHODS: Fifteen white (Norwegian) healthy young adult subjects aged 20-29 years had IOP and CCT measures made, using a non-contact specular microscope and non-contact tonometry (NCT), every hour over a period of 16 h, starting at 06.30 hours. The experiment was repeated on two successive days. RESULTS: The average NCT readings were 12.5 and 11.4 mmHg for days 1 and 2, while pachometry averages were 0.518 and 0.514 mm. The range of NCT values, for any set of readings, averaged 2 mmHg, while this range was 0.014 mm for CCT measures. The reproducibility of these measures, as estimated by the coefficient of variation (COV) was 7.2% for NCT and 1.0% for pachometry measures. The COV for tonometry was inversely proportional to the actual values of these measures. A significant time-related decline in tonometry values was seen over day 1 but not day 2. Pachometry values declined very rapidly over the first hour in both sessions and were essentially constant for the rest of the sessions. The strongest association between pachometry data and NCT measures was noted around midday (12.30 hours) on both sessions. CONCLUSIONS: These studies confirm that the reproducibility (repeatability) of NCT and pachometry are substantially different. It is suggested that this aspect of tonometry needs to be taken into account, especially where there are lower values, when considering the possible impact of corneal thickness on tonometry measures and their interpretation. These types of effects need to be further assessed in older patients with ocular hypertension, or other glaucoma suspects. PMID- 12753486 TI - Basic science and clinical studies coincide: active treatment approach is needed after a sports injury. AB - The basic response to injury at the tissue level is well known and consists of acute inflammatory phase, proliferative phase, and maturation and remodeling phase. Knowing these phases, the treatment and rehabilitation program of athletes' acute musculoskeletal injuries should use a short period of immobilization followed by controlled and progressive mobilization. Both experimental and clinical trials have given systematic and convincing evidence that this program is superior to immobilization - a good example where basic science and clinical studies do coincide - and therefore active approach is needed in the treatment of these injuries. PMID- 12753487 TI - Acute traumatic extension deficit of the knee. AB - The purpose of this study was to evaluate the events leading to acute traumatic extension deficit of the knee and the arthroscopic findings in these patients. A total of 78 consecutive patients treated in the Turku university hospital during the years 1994-1996 were included. The mean annual incidence of acute traumatic extension deficits of the knee in our study was 1.1 per 10 000 inhabitants. The single most common (33%) event causing the extension deficit was non-sports related twisting of the knee. Various sports related activities accounted for 42% of the extension deficits, and soccer was the most common sport in this group. In conclusion, acute traumatic extension deficit of the knee is usually a sign of serious intra-articular damage, and the most likely finding (in 82% of the patients in our study) is either a meniscal rupture, an anterior cruciate ligament (ACL) rupture, a patellar dislocation, or a combination of these. The lesions in these knees require prompt evaluation by an orthopaedic surgeon mainly because of the high number of bucket-handle and menisco-capsular insertion ruptures of the menisci, which are possibly suitable for repair. PMID- 12753488 TI - Recovery of skeletal muscle contractility and hormonal responses to strength exercise after two weeks of high-volume strength training. AB - Exercise induced neuromuscular fatigue, recovery and hormonal responses were studied before (R1) and after (R2) 2 weeks of heavy strength training. Seventeen weight-trained male students were recruited into a heavy training group (HT, n = 10) and a control group (n = 7). During heavy training HT exercised leg extensors every day while control group exercised twice a week. Test workouts (R1 and R2) were used to induce neuromuscular fatigue and hormonal responses. Acute fatigue after the test workouts was reduced after heavy training in the HT group (P < 0.05) but not in the control group. Twenty-two hrs after the test workouts recovery from fatigue was not complete before heavy training, but recovery was complete after heavy training in both groups. The relative change in exercise induced cortisol and GH response, from before to after heavy training, was significantly different between groups, but for both hormones alterations in the control groups response was responsible for the between groups difference. IGF-1 concentration was reduced 22 h after the test workout performed after heavy training in the HT group (P < 0.05). In conclusion, two weeks of high volume strength training attenuated neuromuscular fatigue after a test workout with only minor changes in exercised induced hormone response. PMID- 12753489 TI - Time course of GLUT4 and AMPK protein expression in human skeletal muscle during one month of physical training. AB - Endurance training elicits profound adaptations of skeletal muscle, including increased expression of several proteins. The 5'-AMP activated protein kinase (AMPK) may be one of these, considering the fact that acute exercise increases AMPK activity. Eight young (26 +/- 1 year) lean, healthy males endurance trained one leg (while the other leg remained resting) on an ergometer bicycle for 30 min/day for four weeks (workload corresponding to approximately 70% of maximal oxygen uptake). Muscle biopsies were obtained approximately 18 h after the previous training session. On day eight GLUT4 protein expression was 36% higher in trained (T) compared with untrained (UT) (P < 0.05), but no further increase was seen at day 14 and 30 despite continuously increasing absolute workloads. Expression of AMPKalpha2 and actin did not change with training. In contrast, expression of AMPKalpha1 was 27% higher in T vs. UT muscle (P < 0.05) (measured only on day 30). CONCLUSIONS: GLUT4 protein expression increases substantially after seven days of endurance training with no further increase with prolonged training at progressively increasing workloads. AMPKalpha1 and alpha2 behave differently in their expression in response to endurance training. AMPKalpha1 protein content is increased after one month of training, while no change in AMPKalpha2 and actin expression was detected over the time course of the training period. PMID- 12753490 TI - Soccer injuries in childhood. AB - A prospective study of soccer injuries, involving 287 male youth players, from the ages of 12-15 was conducted in 24 Greek soccer clubs, during the course of one year. A total of 193 players sustained 209 injuries. The incidence of injuries was 4.0 injuries per 1000 h of soccer time per player, and the most common types of injuries were sprains and strains. Surprisingly, it was found that an increase of injury incidences occurred during practice. Fifty-eight injuries required medical assistance. The majority of injuries (80%) were located in the lower extremities. Collision with other players was the most common activity at the time of injury, accounting for 40% of all injuries. The conclusion of this study and the evidence from other studies suggests that youth soccer is a relatively low risk sport. However, a substantial amount of injuries could be prevented. It is necessary to identify the risk factors, which are associated with these types of injuries. PMID- 12753491 TI - Non-dominant leg training improves the bilateral motor performance of soccer players. AB - The aim of this experiment was to evaluate bilateral motor performance effects from training the non-dominant leg of competitive soccer players. The subjects were 39 soccer players, 15-20 years of age, performance-matched and randomly divided into a training group (n = 18) and a control group (n = 21) both belonging to the same team. Both groups were tested by using two standardised foot-tapping tests and three soccer-specific tests. The training intervention consisted of the experimental group participating in all parts of their soccer training except full play, using the non-dominant leg for 8 weeks. Statistical analyses for the soccer-specific tests revealed that the experimental group improved significantly as compared to the control group from the pre-test to the post-test period in their use of the trained non-dominant leg. Somewhat unexpectedly, the experimental group also improved significantly in the tests, which made use of the dominant side. The standardised foot-tapping tests revealed similar results. The results might be explained by improved generalised motor programmes, or from a Dynamic Systems Approach, indicating that the actual training relates to the handling of all the information available to the subject in the situation, and that the body self-organises the motor performance. PMID- 12753492 TI - Maximal oxygen uptake and power of lower limbs during a competitive season in triathletes. AB - BACKGROUND: In order to study the effect of a competitive triathlon season on maximal oxygen uptake (VO2max), aerobic power (AeP) and anaerobic performance (AnP) of the lower limbs, eight triathletes performed exercise tests after: (1) a pre-competition period (Pre-COMP) (2) a competitive period (COMP), and (3) a low (volume and intensity) training period (Post-COMP). The tests were a vertical jump-and-reach test and an incremental exercise test on a cycle ergometer. Ventilatory data were collected every minute during the incremental test with an automated breath-by-breath system and the heart-rate was monitored using a telemetric system. RESULTS: No changes in VO2max were observed, whereas AeP decreased after Post-COMP compared to Pre-COMP and COMP and AnP decreased during COMP compared to Pre-COMP and Post-COMP. In addition, second ventilatory threshold (VT2) and power output at first ventilatory threshold (VT1) and VT2 decreased after Post-COMP. CONCLUSION: This study showed that six weeks of low volume and intensity of training is too long a period to preserve adaptations to training, although a stable maximal oxygen uptake throughout the triathlon season was observed. Moreover, the AnP decrease during COMP was probably in relation with the repetitive nature of the training mode and/or triathlon competitions. PMID- 12753493 TI - Loss of bone mineral of the hip and proximal tibia following rupture of the Achilles tendon. AB - In a prospective uncontrolled study 12 patients suffering from a rupture of the Achilles tendon treated operatively with surgical repair and post-operative immobilization in a short plaster cast for 6 weeks had bilateral measurements of bone mineral content (BMC) of the proximal tibia and bone mineral density (BMD) of the femoral neck and greater trochanter. The measurements were performed by dual energy X-ray absorptiometry (DEXA) and scans were performed post-operatively within 7 days after the operation and with follow up after 6 weeks, 3, 6, and 12 months. In the operated legs, BMC of the proximal tibia showed a progressive decrease reaching a total bone loss of 6.4% (95%-CL: -10.6%; -2.3%) 1 year after the injury. Bone mineral density at the hip of the operated legs also decreased significantly and 1 year after the injury BMD was 2.5% (95%-CL: -5.5%; 0.5%) and 6.8% (95%-CL: -9.8%; -3.7%) below the initial value in, respectively, the femoral neck and greater trochanter. Patients with a previous Achilles tendon rupture must be considered to be some years ahead in their natural osteoporotic process of the bones of the affected legs, and an increased risk of osteoporotic fractures must be considered not to be only theoretical. PMID- 12753494 TI - Optoelectronic measures in the analysis of running target shooting. AB - In this study, we examined the construct validity and practical significance of an optoelectronic shooting training system (Noptel ST 2000 Sport) for the technical analysis of running target shooting. A total of 37 male shooters of three different skill levels participated in the study. Principal component analysis revealed four common factors of 16 variables describing the aiming trajectory of the rifle barrel: (i) Holding area, (ii) Accuracy of aiming, (iii) Cleanness of triggering and (iv) Time on target. These factors were suggested to describe the essential components of running target shooting. According to the discriminant analysis, the shooters of various skill levels seemed to discriminate successfully into three groups when the aiming trajectory data were analysed. Finally, the aiming trajectory variables represented a 43% of the total variance in the shooting score. In summary, the present data indicated that the optoelectronic shooting training system had practical significance and supported the technical analysis of rifle barrel movement in running target shooting. A sub set of variables, which reflect the essential information of running target shooting performance, were outlined for training and coaching purposes. PMID- 12753496 TI - Regulator T cells: specific for antigen and/or antigen receptors? AB - Adaptive immune responses are regulated by many different molecular and cellular effectors. Regulator T cells are coming to their rights again, and these T cells seem to have ordinary alpha/beta T-cell receptors (TCRs) and to develop in the thymus. Autoimmune responses are tightly regulated by such regulatory T cells, a phenomenon which is beneficial to the host in autoimmune situations. However, the regulation of autoimmune responses to tumour cells is harmful to the host, as this regulation delays the defence against the outgrowth of neoplastic cells. In the present review, we discuss whether regulatory T cells are specific for antigen and/or for antigen receptors. Our interest in these phenomena comes from the findings that T cells produce many more TCR-alpha and TCR-beta chains than are necessary for surface membrane expression of TCR-alphabeta heterodimers with CD3 complexes. Excess TCR chains are degraded by the proteasomes, and TCR peptides thus become available to the assembly pathway of major histocompatibility complex class I molecules. Consequently, do T cells express two different identification markers on the cell membrane, the TCR-alphabeta clonotype for recognition by B-cell receptors and clonotypic TCR-alphabeta peptides for recognition by T cells? PMID- 12753497 TI - Cellular mechanism of thymic involution. AB - Involution of the thymus and alterations in the development of thymocytes are the most prominent features of age-related immune senescence. We have carried out a comparative analysis of thymocyte and stroma in rapid thymic involution DBA/2 (D2) strain of mice compared with slow involution C57BL/6 (B6) strain of mice. Analysis of mice at 15 months of age suggested an age-related decrease in the thymocyte cell count, a block in the development of T cells and cortical involution in D2 mice compared with 3-month-old mice. TUNEL (terminal deoxynucleotidyl-transferase-mediated dUTP-digoxigenin nick end labelling) staining and fluorescence-activated cell sorter (FACS) analysis showed that there was a significant increase in apoptotic cells in the cortex region of thymus in 15-month-old D2 mice compared with the same aged B6 mice. The thymocyte proliferation rate, as assessed by bromodeoxyuridine (BrdU) staining and [3H] thymidine incorporation assay, was lower in 3-month-old D2 mice compared with the same age B6 mice. Immunohistochemical staining showed that the arrangement of MTS (mouse thymus stromal)-10+ epithelial cells and MTS-16+ connective tissue staining pattern had become disorganized in 15-month-old D2 mice but remained intact in B6 mice of the same age. These results suggest that, in D2 mice, both the thymocytes and stromal cells exhibit age-related defects, and that the genetic background of mice plays an important role in determining age-related alterations in thymic involution. PMID- 12753498 TI - Analysis of T-cell receptor beta chain variable gene segment usage in healthy adult responders and nonresponders to recombinant hepatitis B vaccine. AB - One to 10 per cent of healthy adult individuals do not produce protective levels of anti-hepatitis B surface (HBs) antibodies, following a standard vaccination protocol. Lack of an HBs antigen (Ag)-specific T-cell repertoire is amongst the possible defects, which may lead to humoral unresponsiveness and is the main objective of this study. We analysed TcR BV (T-cell receptor beta chain variable) gene usage in T lymphocytes from nine healthy adult responders and six nonresponders to recombinant HB vaccine, before and after booster vaccination. CD4+ and CD8+ T-cell populations were isolated from peripheral blood mononuclear cells by magnetic beads, and the expression of TcR BV genes in each population was investigated by reverse transcription polymerase chain reaction and hybridization with specific probe. When the usage of each TcR BV gene within CD4+ and CD8+ T cells of the responders was compared with that of nonresponders, statistically significant difference (P < 0.01) was noted for BV5S2-3 gene family in CD4+ T cells of nonresponders. Furthermore, individual vaccinees were shown to overexpress several TcR BV genes. To characterize the T-cell repertoire and determine their clonal nature, analysis of CDR3 length polymorphism was performed. Our results show that T-cell response to HBsAg is generally oligoclonal and involves multiple BV families. Furthermore, overexpressed individual TcR BV genes and CDR3 length distributions in response to HBsAg are subject-dependent. In conclusion, our results are not in line with the notion that defective TcR repertoire may be an explanation for unresponsiveness to recombinant HBsAg vaccine. PMID- 12753499 TI - The cholera toxin B subunit is a mucosal adjuvant for oral tolerance induction in type 1 diabetes. AB - When conjugated to various proteins, the nontoxic B-chain of cholera toxin (CTB) significantly increases the ability of these proteins to induce immunological tolerance after oral administration. Here, we investigated if a nonconjugated form of CTB enhances the induction of immune tolerance after oral insulin administration. Induction of immunological tolerance was studied after oral administration of insulin preparations in three mouse models; an insulin/ovalbumin coimmunization model, a model of virus-induced diabetes in transgenic RIP-LCMV-NP mice and in nonobese diabetic (NOD) mice serving as a model of spontaneous diabetes. In the immunization model, we demonstrate that mixing with CTB increases the tolerogenic potential of insulin, approximately 10 fold. Titration of the CTB concentration in this system revealed that an insulin : CTB ratio of 100 : 1 was optimal for the induction of bystander suppression. Further studies revealed that this insulin : CTB ratio also was optimal for the prevention of diabetes in a virus-induced, transgenic diabetes model. In addition, the administration of this optimal insulin-CTB preparation significantly prevented the onset of diabetes in old NOD mice with established islet infiltration. The data presented here demonstrate that CTB, even in its unconjugated form, functions as a mucosal adjuvant, increasing the specific tolerogenic effect of oral insulin. PMID- 12753500 TI - Quantification of cytokine gene expression using an economical real-time polymerase chain reaction method based on SYBR Green I. AB - Assessment of cytokine expression has become crucial to understand host responses to infections as well as autoimmunity. Several approaches including Northern blot, RNase protection assay and enzyme-linked immunosorbent assay have been used for this purpose, but they are time consuming, labour intense, and relatively large quantity of the samples is usually required. Recently, a technique termed real-time reverse transcriptase-polymerase chain reaction (RT-PCR) has been developed to determine genetic expression with great sensitivity and specificity; however, specialized instrumentation and costly reagents are usually needed. We aimed at using low-cost reagents for real-time PCR. This was achieved by adapting a conventional RT-PCR protocol to the quantitative real-time format, by the addition of the SYBR Green I reagent. We validated the approach by assessing the cytokine gene expression of murine splenocytes upon stimulation with phorbol 12 myristate 12-acetate (PMA)-ionomycin. The results using this technique were compared with those obtained with the well-established gene array method. We conclude that the use of the SYBR Green I reagent during real-time RT-PCR provides a highly specific and sensitive method to quantify cytokine expression with accuracy and no post-PCR manipulation. PMID- 12753501 TI - The possibility of B-cell-dependent T-cell development. AB - The development of T cells is thought to be independent of B cells. However, defects in cell-mediated immunity in individuals with B-cell deficiency suggest the contrary. To test whether B cells affect T-lymphocyte development, we constructed mice with a monoclonal T-cell compartment (MT) and monoclonal B- and T-cell compartments (MBTs). In these mice, the T cells expressed a DO 11.10 transgenic (DO-T) cell receptor restricted to major histocompatibility complex (MHC) class IId. While CD4+ DO-T lymphocytes are rare in transgenic H-2b MT mice, we found that in H-2b MBT mice under the influence of B cells, DO-T lymphocytes mature into large numbers of CD4+ peripheral T cells. H-2b MBT mice have more CD4+ thymocytes than H-2b MT mice. These data are consistent with the view that B cells play some role in thymocyte development. PMID- 12753502 TI - Cloning, sequencing and expression of immunoglobulin variable regions of murine monoclonal antibodies specific for the P1.7 and P1.16 PorA protein loops of Neisseria meningitidis. AB - The P1.7 and P1.16 epitopes on the PorA protein on the outer membrane of Neisseria meningitidis can induce protective antibodies upon vaccination. Structural analysis of antibodies to these targets can give information on the immune response induced by these epitopes and can reveal any structural similarities among the antibodies. To do so, we have isolated the immunoglobulin (Ig) variable genes from four mouse hybridomas expressing antibodies against the P1.7 and P1.16 epitopes. These V genes were successfully expressed as functional chimeric (ch) mouse/human IgG1 antibodies by subcloning them into expression vectors containing the constant genes of human heavy and light chains. Sequencing the two sets of V genes against P1.16 revealed a high degree of homology, similar to that previously published for P1.7 V genes. The close homology allowed us to interchange heavy and light chains between antibodies in some instances to construct new antibodies that bind the original antigen. This study demonstrates that the immune response in mice against the meningococcal PorA protein epitopes P1.7 as well as P1.16 is limited to few and very similar germline genes, and therefore the P1.7- and P1.16-specific antibodies share high degree of similarities amongst each other. These V genes were used to construct chimeric antibodies with conserved antigen-binding activity. PMID- 12753503 TI - Interleukin-8 production in tuberculous pleurisy: role of mesothelial cells stimulated by cytokine network involving tumour necrosis factor-alpha and interleukin-1 beta. AB - Interleukin-8 (IL-8) plays an important role in the host immune response to Mycobacterium tuberculosis by recruiting inflammatory cells to the site of infection. Here, we investigated the role of pleural macrophages and mesothelial cells in the production of IL-8 in tuberculous pleurisy. Large concentrations of IL-8 were detected in tuberculous pleural effusions, but not in pleural effusions associated with congestive heart failure (CHF). Tuberculous pleural macrophages and M. tuberculosis-infected CHF pleural macrophages produced large concentrations of IL-8. When immunohistochemistry was performed on pleural tissues, antigenic IL-8 was detected in the mesothelial cells lining the tuberculous pleura. Direct stimulation of cultured CHF pleural mesothelial cells with M. tuberculosis induced IL-8 secretion. However, conditioned media from M. tuberculosis-infected pleural macrophages (CoMTB) induced greater mesothelial cell IL-8 secretion. Tumour necrosis factor-alpha (TNF-alpha) and IL-1beta induced mesothelial cell IL-8 mRNA expression, and neutralizing anti-TNF-alpha antibody and IL-1 receptor antagonist nearly completely obliterated CoMTB-induced mesothelial cell IL-8 mRNA expression and protein secretion. These findings demonstrate that both pleural macrophages and mesothelial cells produce IL-8 in tuberculous pleurisy, and cytokines produced by M. tuberculosis-infected macrophages mediate mesothelial cell IL-8 production. PMID- 12753504 TI - Comparison of immunoglobulin heavy chain rearrangements between peripheral and glandular B cells in a patient with primary Sjogren's syndrome. AB - Myoepithelial sialadenitis (MESA) of the major salivary glands is a characteristic feature of primary Sjogren's syndrome (pSS). To delineate systemic and organ-specific influences on B cells in a patient with pSS and benign MESA, individual B cells were simultaneously obtained from the peripheral blood and inflamed parotid gland. Immunoglobulin variable heavy chain (VH) rearrangements in single sorted CD19+ B cells were subsequently amplified, sequenced and analysed. Despite the presence of two clonal expansions using VH1-08 and VH2-70 segments, respectively, the majority of glandular B cells were polyclonal, resembling the VH gene usage and mutational pattern of the corresponding blood population. However, striking differences were observed in the proportion of cells expressing mutated VH rearrangements (blood, 28.9% versus parotid, 80.4%; P < 0.0001). Moreover, the glandular productive VH rearrangements differed significantly from their blood counterparts by a higher mutational frequency (P < 0.0001), shorter CDR3 lengths (P = 0.001) and a less frequent usage of JH6 (P = 0.0292), indicating an accumulation of memory B cells in the inflamed parotid. Thus, both preferential influx/homing of memory B cells and local proliferation may contribute to the pattern of benign MESA in pSS. Notably, one of the glandular clonal rearrangements (using VH1-08) was also detected in the patient's peripheral repertoire. PMID- 12753505 TI - Interferon-gamma receptor-1 gene polymorphism in tuberculosis patients from Croatia. AB - Recent studies have indicated that the interleukin-12/interferon-gamma (IFN gamma) axis is important in mycobacterial infection susceptibility. Using an intronic (CA)n polymorphic microsatellite marker within the IFN-gamma receptor-1 (IFNGR1) gene, we have compared the allelic frequencies of this marker in hospitalized tuberculosis patients (n = 120) with that of controls (n = 87) from Rijeka, Croatia. We identified 13 (CA)n alleles in the tuberculosis patients, whereas only 10 were found in the controls. A significant difference between one allelic marker and the control group was observed (P = 0.02, 95% confidence interval 0.14-0.94), suggesting a possible protective association. In contrast, several other allelic markers showed a trend towards association with the disease. We also found a trend towards an increased frequency in homozygosity of one allelic marker in patients (11.7%) as compared with controls (4.6%). We conclude that there is no evidence for disease association of the IFNGR1 gene marker in Mendelian-type (single-allele) inheritance. However, our results also suggest that unidentified allelic variations in the IFNGR1 gene might elevate or decrease the risk in this ethnic population, as a part of the multigenic predisposition to tuberculosis. PMID- 12753506 TI - T-cell receptor excisional circles, telomere length, proliferation and apoptosis in peripheral blood mononuclear cells of human immunodeficiency virus-infected individuals after 18 months of treatment induced viral suppression. AB - This study evaluated the effect of highly active antiretroviral therapy (HAART) induced viral suppression on T-cell receptor excisional circles (TRECs), telomere length, proliferative responses and spontaneous as well as phytohaemagglutinin (PHA)-stimulated lymphocyte apoptosis in 27 human immunodeficiency virus (HIV) infected individuals followed for 18 months during HAART. Our results show that HAART significantly increased the level of TRECs in CD4+ cells (P = 0.003) after 18 months of almost continuously suppressed HIV-RNA levels. Lymphocyte proliferative responses and apoptosis levels in patients were significantly lower and significantly higher, respectively, compared with healthy controls. The proliferative response and apoptosis levels did not change during follow up. Changes in telomere length were observed in CD4+ and in CD8+ T cells. The study demonstrated that HAART induces normal TREC levels in the CD4+ T-cell pool. However, the other perturbed functions in T cells indicate that immune reconstitution is incomplete and may need longer viral suppression. PMID- 12753507 TI - Increased brain-derived neurotrophic factor expression in white blood cells of relapsing-remitting multiple sclerosis patients. AB - Central nervous system (CNS)-autoreactive immune responses can exert neuroprotective effects, possibly mediated via the release of neurotrophic factors from infiltrating leucocytes. Herein, we analysed neurotrophin and cytokine mRNA levels using TaqMan polymerase chain reaction in unstimulated peripheral blood mononuclear cells (PBMCs) from multiple sclerosis (MS) patients in remission and controls. We demonstrate that mRNA for brain-derived neurotrophic factor (BDNF), but not neurotrophin-3 or nerve growth factor (NGF), is readily detectable in PBMC and that levels in MS are increased by approximately 60% compared with patients with other neurological diseases or healthy subjects. These results provide additional evidence that a potentially neuroprotective facet of autoimmune inflammation is present in MS. PMID- 12753508 TI - Expanding knowledge in family health care. PMID- 12753509 TI - Maternal adaptation during childbearing in Norway, Sweden and the United States. AB - The purpose of the study of 147 healthy primiparous women is to investigate the relationship between maternal adaptation during pregnancy and postpartum. This report is part of a larger study of relationships between family dynamics and maternal adaptation during childbearing in Norway, Sweden and the US. Variables included maternal adaptation, age, social status, mother's employment, type of birth, length of hospitalization and mother infant time together in the hospital. Mothers reporting greater adaptation during pregnancy reported greater adaptation postpartum. Mothers who believed themselves adapted indicated that they were better prepared for labour, had more control over their birth experience, a better relationship with their partner, and perceived greater participation in child care from their partner. Few differences were found in maternal adaptation among all mothers, and few relationships were found among selected sociodemographic variables and maternal adaptation. Formation of maternal identity and mother's confidence in her ability to cope with the tasks of motherhood was affected by mothers' prenatal identification with the motherhood role, supporting the notion that knowing what to expect provides a sense of control. Mothers' prenatal identification with the motherhood role directly or indirectly affected her satisfaction with motherhood. Nurses may want to develop practice strategies to support these findings. PMID- 12753510 TI - Severe postpartum distress in Icelandic mothers with difficult infants: a follow up study on their health care. AB - This article describes help seeking and health care of mothers with a difficult infant who suffered long-term depressive symptoms and a high degree of parenting stress. A subsample of severely distressed mothers (n = 37) was recruited from a cross-sectional national survey and followed up 2 months later by a semi structured telephone interview. The survey included all Icelandic women who gave birth during a quarter of a year and had a live baby 2 months later (n = 1053). All distressed mothers with a difficult infant were selected from sample respondents on preset scores of two self-report distress measures. The study sample emerged during the selection process for an intervention study from which it was excluded on grounds of very high distress scores. Results showed that 5% of the surveyed population were postpartum severely distressed. Findings from this follow-up study revealed that only one woman of four received health care for severe distress by various professionals. One woman of six received help from others. Very few women utilized the services available at Health Care Centers. About half of the women held attitudes that hindered them in seeking help and health care. It is concluded that postpartum severely distressed women receive little primary health care for mental health problems and the majority of them show little initiative to seek out for help. More active outreach by health professionals is recommended. PMID- 12753511 TI - Vulnerability of family caregivers in terminal palliative care at home; balancing between burden and capacity. AB - This paper reports on a grounded theory interview-based study with 13 family members aged 28-80 years caring for terminally ill people at home (with a life expectancy of 3 months or less) in the Netherlands. The project was approved by the ethics committee of the Maastricht University Hospital. The aim of this study was to explore the experiences of family caregivers, their needs for home care, and which health services they receive. Data were analysed using the constant comparative method. 'Vulnerability' was identified as the core category. Caring for a terminally ill person at home requires continuous balancing between care burden and capacity to cope. Whether or not the carer will succeed in keeping in optimum balance is dependent on a number of factors impinging on the caregiver's vulnerability. Care burden, restricted activities, fear, insecurity, loneliness, facing death, lack of emotional, practical and information-related support were identified from the data as factors having the potential to increase the caregiver's vulnerability, and may be risk factors for fatigue and burnout. Continuing previous activities, hope, keeping control, satisfaction and good support are factors which may decrease the caregiver's vulnerability, and may protect against fatigue and burnout. The experiences of the caregivers in our study showed that the support from informal and professional caregivers was not sufficient. Education and practical tools may make professionals more sensitive for the vulnerable position of family caregivers, even when these caregivers do not show their vulnerability. PMID- 12753512 TI - Strength of motherhood: nonrecurrent breast cancer as experienced by mothers with dependent children. AB - The experience of breast cancer in mothers of dependent children and the strategies these women use to handle their situation of illness in relation to the children was the focus of this study. Ten women with nonrecurrent breast cancer were interviewed using phenomenology as a theoretical framework. The findings reveal that the women needed to balance themselves from being needed and perhaps not existing, balancing between demands, a will to be strong and allowing herself to be sick, and balancing between telling the truth and protecting the children from the truth. The women had strategies to help them in this struggle. To carry on as usual and continue everyday life became very important to them. To do this they had to combine their own strength with support from others and try to turn things into a positive perspective. The essential meaning of living with breast cancer and having dependent children implied using the strength of motherhood to balance conflicting forces and thereby continuing everyday life. The clinical implications include awareness and support to maintain everyday routines and generate strength. PMID- 12753513 TI - Adolescent subjective well-being and family dynamics. AB - The purpose of this study was to examine the relationships between adolescent subjective well-being (SWB) and family dynamics perceived by adolescents and their parents. A sample of 239 pupils (51% female) from seventh and ninth grades completed the Berne questionnaire of SWB (youth form), two subscales from an original Finnish SWB scale and the Family Dynamics Measure II, and one of their parents (n = 239) filled in the Family Dynamics Measure II. Results indicated that parents assessed family dynamics better than did their adolescent child. Furthermore, there was no association between family dynamics perceived by adolescents and family dynamics assessed by one of their parents or between the adolescent SWB and parental perception of family dynamics. Multiple stepwise regression analysis indicated that certain aspects of family dynamics perceived by adolescents were related to adolescent global satisfaction and ill-being. Specifically, adolescents' perception of high level of mutuality and stability in the family as well as male gender and lack of serious problems in family were predictors of adolescent global satisfaction. Furthermore, disorganization in the family and poor parental relationship perceived by adolescents, being female, serious problems and illness in family predicted a high level of adolescent global ill-being. PMID- 12753514 TI - Functioning of child maltreating families: lack of resources for caring within the family. AB - The purpose of this study was to develop a preliminary theoretical model of functioning of child maltreating families. Child maltreatment was defined as physical, psychological or sexual abuse or neglect caused by the child's primary caregivers. The aim was to generate knowledge needed in developing understanding and the care of these families. Data were collected using interviews with open ended questions from child maltreating families (n = 20). Transcribed interviews were analysed by using grounded theory and its constant comparative method. The categories of family functioning included structural factors of the family, relationships between family members, relationships outside the family, family strengths, and risk factors and manifestations of maltreatment. The relationships that emerged between these categories included conflicts and negative circles of family life and the effects of child maltreatment on the family. Lack of resources for caring within the family emerged as the core category which described the phenomenon in general and was connected to all other categories of family functioning. The findings can be useful in developing the care of child maltreating families, in nursing education and in preventing child maltreatment. PMID- 12753515 TI - Women's worries during pregnancy: testing the Cambridge Worry Scale on 200 Swedish women. AB - The Cambridge Worry Scale (CWS) is an instrument including 16 items measuring women's major worries during pregnancy. The aim of the study was to test the scale, translated into Swedish, on pregnant women in Stockholm. We also wanted to explore whether these women were worried about any item not included in the scale. An additional aim was to study possible variation in women's worries related to gestational week. Two hundred women were recruited. The average age was 31 years and 56% were primiparas. Gestational age ranged from 8 to 42 weeks, with a median of 28 weeks. The reliability of the scale was satisfactory (Cronbach's alpha coefficient 0.81). The major worries were about the baby's health, giving birth and miscarriage. These items, all related to pregnancy outcomes, were followed by worries about financial matters. An additional concern not included in the scale was about the maternity services in Stockholm, i.e. that the hospital would be overbooked, the staff being too busy or the medical safety not being guaranteed. Few women worried about their relationship with their partner or if he would be present at birth. Some of the items showed a pattern with a period of less worry in midpregnancy. PMID- 12753516 TI - Parents' needs to talk about their experiences of childbirth. AB - The purpose was to describe parents' experiences of childbirth and their views about having a postpartum talk. Questionnaires were distributed to parents (350 mothers and 343 fathers) in maternity wards in one hospital in Stockholm during a 4-week period in 1999. Questionnaires from 251 mothers and 235 fathers were analysed. Preparation for childbirth, care received during delivery, and experiences of labour and delivery are described and analysed against whether a postpartum talk, was wanted or not. Comparisons between first-time mothers/fathers and multiple-time mothers/fathers were made. The results show that 66% of first-time mothers, 74% of multiple-time mothers, 58% of first-time fathers, and 30% of multiple-time fathers wanted to talk about the delivery. The issues, which the parents considered should be included in such a postpartum talk were the birth process, normal/complicated delivery, feelings of failure, pain and pain relief. Parents mainly wanted to talk to the midwife who delivered the woman and the best time for the postpartum talk seems to be at the maternity ward before discharge. PMID- 12753517 TI - The view of the child health nurse among mothers. AB - The aim of this exploratory study was to investigate mothers' experiences of their encounters with the child health (CH) nurse. A cross-sectional design was used for the study, based on data from 140 mothers gathered by the critical incident technique. The analysis was accomplished by a thematic content analysis, using inductive reasoning in three steps. Symbolic interactionism was used as a frame of reference. The results suggest that the central factor in the encounter between mother and nurse is that they are able to share the realm of motherhood, meaning that the nurse is open and willing to share all types of emotions, experiences and attitudes related to being a mother. Given this basis, other important factors are the supply of sound advice and practical interventions, and that the nurse is reassuring and accessible. The majority of the participating mothers had experienced CH nurses who had provided them with valuable support during troublesome incidents. However, there were also several dissatisfied mothers who had expected support but thought they received insulting treatment instead. The mothers and the nurses have varying experiences and background and therefore different perspectives, which may lead to difficulties in understanding each other. Knowledge about the important factors, that affect the mother-nurse encounter, can be used to strengthen the nurses' positive behaviours and facilitate understanding of how disappointed mothers have experienced their health care encounters. PMID- 12753518 TI - Supportive and nonsupportive qualities of child health nurses' contacts with strained infant mothers. AB - AIMS: To examine how strained mothers perceive the support they receive from Swedish child health nurse. POPULATION AND METHODS: Twenty-four consecutive strained, infant mothers at nine child health centres were invited to participate. The mothers were interviewed when the child was 8 months of age. The mothers also reported their perceptions of their parenting competence by means of a questionnaire. RESULTS: Five components of the nurses' support were discerned: positive emotional support, positive informational support, support of the parental role which was either positive or negative, and pressure to accept social norms on child care. A third of the mothers reported strong positive support, decreasing depressive symptoms and favourable perceptions of their own parenting ability. Another third of the mothers reported negative contacts with the nurses, persisting depressive symptoms, and unfavourable perceptions of their parenting competence. The nurses' pressure for the mothers to adapt to certain social norms of child care seemed to affect the latter group of mothers negatively. CONCLUSIONS: In order to support strained mothers effectively it is necessary to clarify and resolve conflicts between different programme objectives. PMID- 12753519 TI - An examination of the needs of parents of hospitalized children: comparing parents' and staff's perceptions. AB - The needs of parents of hospitalized children have received some attention in the health literature, but few studies have compared parents' perceptions of needs with staff's ideas about parents' needs. The aim of this study was to examine differences between the perceptions of the needs of parents of hospitalized children held by staff - nurses, doctors and allied health staff, and parents in a 150-bed paediatric hospital in Sweden. The convenience sample comprised 132 staff - nurses, doctors and allied health staff and 115 parents of children admitted to all the wards except intensive care. Kristjansdottir's "needs of parents of hospitalized children" questionnaire (NPQ) was the instrument of choice and was modified slightly for use with staff. Results indicated significant differences in perceptions of the importance of different needs of parents, of how well they were being met in the hospital and how much help the parents needed to have them filled. Differences between parents' and staff's perceptions of the importance of parental needs were found in areas relating to psychosocial needs, but in general, in that hospital, the needs were being adequately met. The main differences between staff's and parents' results were in the degree of independence shown by parents in requiring help to have their needs met. This demonstrates either that parents are much more independent than appraised by staff, or, that parents are sometimes unaware of the level of assistance available. PMID- 12753520 TI - Nurses' experiences of interactions with family members in intensive care units. AB - The purpose of this study was to describe nurses' experiences of interactions with family members in intensive care units. Ten experienced Registered Nurses were interviewed, and the interview text was qualitatively analysed by its content. Nursing of families was experienced as an essential, necessary and demanding task, and no systematic assessment and intervention with families were talked about. Two categories emerged from nurses' descriptions: inviting and noninviting interactions between nurses and family members. Inviting interactions were considered when family members were seen as important in the nursing care. The nurses used themselves as instruments to create contact and felt confident working with the family members. Thereby the nurses were forced to reflect on their way of caring and received positive responses from family members. In noninviting interactions, medical and technical tasks were considered to be most important and the nurses considered themselves as experts. They expressed having little time for family members and described being afraid of coming too close to them and having problems with creating relationships. Further research, including direct observations of interactions between nurses and family members, is needed. PMID- 12753521 TI - Family-nurse co-construction of meaning: a central phenomenon of family caring. AB - The purpose of the study was to understand and interpret caring in the family health experience by exploring the interactional phenomenon of family-nurse co construction of meaning in the paediatric intensive care unit (PICU). A hermeneutic phenomenological method within a framework of existentialism and symbolic interactionism was used in the investigation. The convenience sample for this study was four family-nurse dyads, that is four families of critically ill children (all with positive outcomes) and the four nurses assigned to their care who were participating in a larger study. Data were derived from semi-structured interviews regarding significant interactions throughout the child's illness and subsequent significant interactions of families with other nurses and nurses with other families. Trustworthiness of the study was addressed through the criteria of credibility, dependability, transferability and confirmability. Co construction of meaning in the family health experience was found to have two dimensions: interdependent and independent. Both families and nurses described being like family as an essential component of the interdependent experience. Independent dimensions for families were journeying through troubled waters of learning the meaning of the illness event and sensing family comfort through the nurse's care. Independent dimensions described by nurses were journeying through troubled waters of learning to care for families and living with another's fear. The family-nurse interaction, the relational connection and the evolution of meanings that families and nurses construct, was affirmed as the major vehicle in the co-construction experience. Family caring is influenced by the existential meaning constructing, process-oriented, interactional nature of the family health experience. Caring in the family health experience is enhanced through actions the nurse performs on behalf of, and with, the family while understanding the family's unique situation. Caring enacted by nurses in participation with families holds abundant potential for enhancing the family health experience and honor the ethic of caring as central to nursing. PMID- 12753524 TI - Caring about tuberculosis: IUATLD's continuing contributions. PMID- 12753525 TI - Women and tobacco. AB - Smoking prevalence is lower among women than men in most countries, yet there are about 200 million women in the world who smoke, and in addition, there are millions more who chew tobacco. Approximately 22% of women in developed countries and 9% of women in developing countries smoke, but because most women live in developing countries, there are numerically more women smokers in developing countries. Unless effective, comprehensive and sustained initiatives are implemented to reduce smoking uptake among young women and increase cessation rates among women, the prevalence of female smoking in developed and developing countries is likely to rise to 20% by 2025. This would mean that by 2025 there could be 532 million women smokers. Even if prevalence levels do not rise, the number of women who smoke will increase because the population of women in the world is predicted to rise from the current 3.1 billion to 4.2 billion by 2025. Thus, while the epidemic of tobacco use among men is in slow decline, the epidemic among women will not reach its peak until well into the 21st century. This will have enormous consequences not only for women's health and economic wellbeing but also for that of their families. The health effects of smoking for women are more serious than for men. In addition to the general health problems common to both genders, women face additional hazards in pregnancy, female specific cancers such as cancer of the cervix, and exposure to passive smoking. In Asia, although there are currently lower levels of tobacco use among women, smoking among girls is already on the rise in some areas. The spending power of girls and women is increasing so that cigarettes are becoming more affordable. The social and cultural constraints that previously prevented many women from smoking are weakening; and women-specific health education and quitting programmes are rare. Furthermore, evidence suggests that women find it harder to quit smoking. The tobacco companies are targeting women by marketing light, mild, and menthol cigarettes, and introducing advertising directed at women. The greatest challenge and opportunity in primary preventive health in Asia and in other developing areas is to avert the predicted rise in smoking among women. PMID- 12753526 TI - Respiratory health effects of exposure to environmental tobacco smoke. AB - Tobacco smoke is a major component of indoor air pollution. Exposure to environmental tobacco smoke (ETS) is prevalent worldwide despite growing awareness of its adverse health effects on non-smokers. ETS contains the same toxic substances as identified in mainstream tobacco smoke. Cotinine (a metabolite of nicotine) can be measured in urine and serum of non-smokers exposed to ETS and reflects the degree of exposure. In children, exposure to ETS leads to reduced lung function, increased risk of lower respiratory tract illnesses, acute exacerbation of asthma resulting in hospitalization, increased prevalence of non allergic bronchial hyperresponsiveness, increased risk for sudden infant death syndrome (SIDS) and possibly increased risk for asthma. Exposure to ETS is responsible for excess cost to the family's financial resources and demands on health services. In adults, exposure to ETS is associated with increased risk of lung cancer, particularly in those with high exposure and acute and chronic respiratory symptoms that improve after the cessation of exposure. Healthcare providers should advocate for non-smokers' rights in the community and support legislation to limit tobacco exposure. PMID- 12753527 TI - The role of dendritic cells in immune regulation and allergic airway inflammation. AB - Dendritic cells (DC) are potent antigen presenting cells that display an extraordinary capacity to present antigen to naive T-cells and initiate primary immune responses. In the context of the lung and upper airway it is clear that DC play a key role in the regulation of adaptive immune responses to inhaled antigen. DC are particularly sensitive to signals derived from microbes, allergens and the airway tissue microenvironment. By the nature of the signals they provide at the time of antigen presentation, DC can polarize naive T-cells into either T-helper type 1 (Th1) or Th2 effector cells, and are increasingly recognized as having a central role in the establishment of T-cell memory and peripheral immune tolerance. DC form a network within the upper airway and lung, and are rapidly recruited from the circulation in response to a variety of proinflammatory stimuli. Studies using animal models have highlighted the role of DC in both the initiation and maintenance of allergic airway inflammation. In early childhood, human DC are functionally immature, and this is thought to contribute to the development of allergic sensitization in those children who are genetically at risk for the development of atopy. Increased numbers of airway mucosal DC are found in both allergic rhinitis and asthma, while studies of blood derived DC have emphasized important differences between the function of DC from atopic and normal individuals. This article reviews recent information on the involvement of DC in allergic airway disease, and the mechanisms by which DC could be exploited as targets for therapy in asthma and allergic rhinitis. PMID- 12753528 TI - Bronchoalveolar lavage fluid cells in mixed connective tissue disease. AB - OBJECTIVE: Patients with mixed connective tissue disease (MCTD) exhibit clinical features of systemic lupus erythematosus (SLE), systemic sclerosis (SSc), and polymyositis and dermatomyositis (PM-DM). The objective of this study was to clarify differences in BAL findings and immunophenotypes of BAL fluid (BALF) cells of patients with interstitial lung disease associated with these diseases. METHODOLOGY: We were unable to recruit a sufficient number of SLE patients with lung disease. We compared immunophenotypes of lymphocytes and alveolar macrophages (AM) in BALF of 20 MCTD patients with those of 21 SSc and 27 PM-DM patients and tested the relationships between immunophenotypes and pulmonary function in MCTD. RESULTS: MCTD patients had a significantly higher CD4/CD8 ratio with more CD4 positive lymphocytes than PM-DM patients (P = 0.025). In AM phenotypes, MCTD patients had a significantly lower percentage of CD71 positive AM compared with SSc patients (P = 0.023). DLCO was negatively related to absolute numbers of CD8 positive lymphocytes (R = -0.517, P= 0.033). CONCLUSIONS: CD4 positive lymphocytes in BALF were increased in MCTD compared to PM-DM patients, while CD71 positive AM were decreased in MCTD compared to SSc patients. CD8 positive lymphocytes correlated negatively with DLCO measurements in MCTD patients. PMID- 12753529 TI - A marked decrease in L-selectin expression by leucocytes in infants with Bordetella pertussis infection: leucocytosis explained? AB - OBJECTIVE: Infants with Bordetella pertussis infection (whooping cough) have an unexplained lymphocytosis and leucocytosis characterized by an increase in small lymphocytes with convoluted and cleaved nuclei. To characterize these cells immunophenotyping using multiparameter flow cytometry was performed on leucocytes from a group of 11 infants aged 3-6 months with proven pertussis and from uninfected control subjects. METHODOLOGY: The panel of monoclonal antibodies used to elucidate leucocyte subtypes included activation, adhesion, costimulatory, memory, T-helper (Th) 1 and Th2 markers. RESULTS: Patients with pertussis showed an increase in absolute numbers of neutrophils, monocytes, T lymphocytes (both CD4 and CD8), B lymphocytes (including CD10+/CD19+ haematogones) and natural killer (NK) cells. All leucocyte subgroups showed a marked decrease in L-selectin (CD62L) expression. The expression of other adhesion molecules CD11a, CD44 and CD54 on all leucocyte subgroups was unchanged. Expression of costimulatory molecules, CD49D and CD28 on T cells and CD80 and CD86 on monocytes, was unchanged. Lymphocyte activation markers CD69, CD25 and HLA-DR were unchanged. There was an increase in CD45RA+/CD45RO+/CD4+ cells (activated) and CD62L /CD45RO+/CD4+ cells (Th1-like) but no increase in CD7-/CD4+ T cells (Th2-like). CONCLUSIONS: L-Selectin expression mediates extravasation of leucocytes into tissues and is important for homing of peripheral blood lymphocytes to lymph nodes. The significant down-regulation of L-selectin on leucocytes in pertussis infection may prevent leucocyte migration to areas of infection and homing and adhesion of T and B cells to peripheral lymphoid tissues. The increase in lymphocytes with Th1 phenotype may be required for effective immune response to the infective organism. These data provide a possible explanation for the absolute leucocytosis observed in this disease. PMID- 12753530 TI - The significance of tumour markers as an indication for mediastinoscopy in non small cell lung cancer. AB - OBJECTIVE: The purpose of this study was to verify the significance of tumour markers as indicators for mediastinoscopy in non-small cell lung cancer. METHODOLOGY: In the past 4 years, 205 patients with non-small cell lung carcinoma (NSCLC) underwent surgical resection at Chiba Cancer Center, Chiba, Japan. The correlation between the serum levels of eight tumour markers (CEA, AFP, CA19-9, SCC, NSE, CA125, CYFRA, ProGRP) and the presence of N2 disease was analysed. Univariate and multivariate analyses were performed to determine the relationship between both marker levels and clinical findings and N2 disease. RESULTS: In multivariate analysis, positive CEA was significantly associated with the diagnosis of N2 disease. We also demonstrated that when CA125, CYFRA and ProGRP were positive, they were individually significantly associated with N2 disease. However, CEA was superior to the other markers and equivalent to a combination of various tumour markers. CONCLUSION: It was concluded that evaluation of CEA in addition to CT is of use in the diagnosis of N2 disease in NSCLC patients and should be used as an indication for mediastinoscopy. PMID- 12753531 TI - Sputum eosinophilia and bronchial responsiveness in patients with chronic non productive cough responsive to anti-asthma therapy. AB - OBJECTIVE: We aimed to examine airway inflammation and bronchial responsiveness in patients with chronic non-productive cough responsive to anti-asthma therapy. METHODOLOGY: Bronchial responsiveness to methacholine as well as the number of inflammatory cells and concentration of eosinophil cationic protein (ECP) in induced sputum were measured in 42 patients with chronic non-productive cough of unknown origin. Their response to bronchodilator, antiallergic and inhaled or oral glucocorticoid therapy was subsequently assessed. RESULTS: Complete remission of coughing was attained with anti-asthma therapies in 34 patients (responder group), while eight patients did not respond (non-responder group). Twenty patients in the responder group and three in the non-responder group showed bronchial hyperresponsiveness (BHR). The number of eosinophils and ECP levels in the sputum from responders with BHR were significantly increased when compared with those from non-responders and healthy subjects. These sputum measures were also significantly increased in responders without BHR when compared with healthy subjects. However, there were no significant differences in these inflammatory markers between the responders with and without BHR. The neutrophil numbers in the sputum from non-responders and responders both with and without BHR were also significantly higher than in control subjects, but there were no significant differences. CONCLUSIONS: These findings suggest that patients with chronic non-productive cough responsive to anti-asthma therapy characteristically have eosinophilic airway inflammation, which may play an important role in the development of chronic cough. Furthermore, the evaluation of not only bronchial responsiveness but also airway inflammation by examination of induced sputum may be useful for diagnosis and deciding on therapeutic strategies. PMID- 12753532 TI - Secretory leukocyte proteinase inhibitor, alpha-1-antitrypsin deficiency and emphysema: Preliminary study, speculation and an hypothesis. AB - OBJECTIVES: This study investigated (i) whether adequate concentrations of secretory leukocyte proteinase inhibitor (SLPI) in the lungs of alpha-1 antitrypsin (A1AT) deficient patients can explain the variability in the development of emphysema in these individuals, and (ii) whether cigarette smoking jeopardises the protective screen provided by functional SLPI. METHODOLOGY: Four subjects [two normal proteinase inhibitor M (PiM), two abnormal PiZ] were selected from patients presenting for diagnostic bronchoscopy and lung function testing (spirometry, DLco). Each subject underwent BAL and had blood taken for A1AT and SLPI estimation. RESULTS: As expected serum and BAL A1AT concentrations were within the normal range in the normal PiM subjects. In normal subjects, SLPI concentrations in serum and BAL were within the normal range. A1AT-deficient subjects had reduced serum and BAL levels of A1AT reflecting their genetic disorder but showed increased concentrations of SLPI in BAL and serum. Percentage neutrophil elastase (NE) inhibitory capacity of BAL fluid was low in both A1AT deficient subjects and a cigarette-smoking normal subject. In contrast, the NE inhibitory capacity for the normal subject who had never smoked was normal. CONCLUSIONS: These findings suggest that in A1AT deficiency there may be a compensatory increase in SLPI. This may protect the lung against the development of emphysema in A1AT-deficient individuals. Cigarette smokers may have a lower SLPI concentration than non-smokers. This provides an explanation for at least some of the observed variation in the development of emphysema in A1AT deficient subjects. PMID- 12753533 TI - Effect of osmolality on mucociliary transportability and rheology of cystic fibrosis and bronchiectasis sputum. AB - OBJECTIVE: Water is the main constituent of mucus, and its concentration is likely to be important in all aspects of mucus function, including ciliary clearance. The objective of this study was to investigate the effect of water content and osmolality of the mucus on mucociliary transportability. METHODOLOGY: Rheology and ciliary transportability of 10 sputum samples that had been subjected to various manipulations were measured using a mucus-depleted bovine trachea model. RESULTS: It was shown that addition of sodium chloride 0.2 Osmoles/kg (0.585% weight for weight) increased the transportability by 41% (P < 0.01). Evaporation of the sputum to 50% of its original weight caused a 118% increase in transportability (P < 0.0006), but iso-osmolal removal of 50% of the liquid with filter cards led to a non-significant, 25% increase in transportability. Parallel plate viscoelasticity was approximately doubled in both the evaporated and liquid-depleted samples, but was not changed by the addition of 0.2 Osmoles/kg of sodium chloride. The correlation between the osmolality of sputum and ciliary transportability (r = 0.54, P= 0.005) was better than the correlations between the viscosity (r = 0.21, P= 0.27) or elasticity (r = 0.23, P= 0.23) and ciliary transportability. CONCLUSIONS: These results suggest that the osmolality of sputum exerts a greater influence on mucociliary clearance than its viscoelastic properties. PMID- 12753534 TI - Influence of stressful life events on the onset of sarcoidosis. AB - OBJECTIVE: The onset of sarcoidosis may be triggered by any hereditary and/or environmental factor. Among these factors, psychosocial stress may play a critical role in the onset of sarcoidosis. The aim of the present study was to evaluate the influence of stressful life events on the onset of sarcoidosis. METHODOLOGY: The social and/or life events experienced prior to diagnosis in 55 patients with sarcoidosis, were evaluated quantitatively using the Social Readjustment Rating Scale (SRRS), the most authoritative method to quantify the magnitude of the events requiring changes in lifestyle. In addition, personality with respect to stress-reactivity was simultaneously evaluated using the Minnesota Multiphasic Personality Inventory (MMPI) alexithymia scale. Both the degree of stress, evaluated by SRRS, and stress-reactivity, evaluated by MMPI scale, of sarcoidosis patients were compared with those of 92 healthy controls. RESULTS: The magnitude of stressful life events was significantly higher in patients with sarcoidosis compared with healthy controls. In addition, capacity for coping with stress was found to be inferior in sarcoidosis patients compared with that in the control groups. CONCLUSION: These results indicated that psychosocial stress assessed on the basis of alexithymic characteristics of the self perception of stress may be partly implicated in the development of sarcoidosis. PMID- 12753535 TI - COPD prevalence in 12 Asia-Pacific countries and regions: projections based on the COPD prevalence estimation model. AB - OBJECTIVE: COPD is a leading cause of mortality and morbidity worldwide. Despite the high rates of cigarette smoking, and the wide use of biomass fuels, there is very little objective data on the prevalence of COPD in Asia. METHODOLOGY: We used a COPD prevalence model to estimate the prevalence of COPD in 12 Asian countries. This model is a validated, computerized tool that uses epidemiological relationships and risk factor prevalence to project the prevalence of COPD within a given population aged 30 years and older. RESULTS: The total number of moderate to severe COPD cases in the 12 countries of this region, as projected by the model, is 56.6 million with an overall prevalence rate of 6.3%. The COPD prevalence rates for the individual countries range from 3.5% (Hong Kong and Singapore) to 6.7% (Vietnam). CONCLUSIONS: The COPD prevalence rates projected by the model reflect the high prevalence of the risk factors for the disease in Asia. The combined prevalence of 6.3% for these countries is considerably higher than the overall rate of 3.8% as extrapolated from WHO data for this region. These estimates highlight the need for further epidemiological studies to support appropriate allocation of resources for the prevention and management of COPD. PMID- 12753536 TI - A step-wise application of methylprednisolone versus dexamethasone in the treatment of acute exacerbations of COPD. AB - OBJECTIVE: The aim of the study was to explore the clinical value of a step-wise application of methylprednisolone (MP) compared to dexamethasone (DXM) in acute exacerbations of COPD. METHODOLOGY: One hundred and forty-two patients with an acute exacerbation of COPD were divided randomly into two groups: 71 patients were treated with MP and the other 71 patients were treated with DXM. Otherwise each group was given the same basic treatments: antibiotics, bronchodilators, oxygen therapy as well as standard hospital care. The patients in the MP group were given a tapering dose of MP for 7-14 days, and the patients in the DXM group were given a corresponding tapering dose of DXM for 7-14 days. Then both groups were given a gradually reducing dose of oral prednisone for 2-3 weeks. Two weeks before the prednisone was tapered off, inhaled corticosteroid was introduced. The patients' symptom scores, physical signs, per cent predicted FEV1%, and arterial blood gases were monitored before treatment and after the seventh day of treatment. RESULTS: There was an obvious improvement in symptoms after 1-3 days in all 71 patients in the MP group, with their wheezing being distinctly reduced or disappearing entirely. The maximum benefit that occurred in the MP group (90.14%) was considerably higher than that of the DXM group (25.35%), P < 0.05. The predicted FEV1% in the MP group increased from 46.7 +/- 10.6 to 67.5 +/- 12.4, compared with an increase in the DXM group from 50.1 +/- 7.6 to 58.9 +/- 10.8. The difference between the two groups was significant (P < 0.05). CONCLUSIONS: An adequate and tapering dose of MP used in acute exacerbations of COPD can relieve the inflammatory reaction in airways and reduce airway spasm more promptly than DXM. PMID- 12753537 TI - Prescribing and administration of nebulized bronchodilators: a prospective audit in a university hospital. AB - OBJECTIVE: Inaccurate and incomplete prescribing of nebulized bronchodilators can result in uncertainty and suboptimal treatment. A prospective study was carried out to assess the completeness of prescription and the quality of bronchodilator drug administration via nebulizers. METHODOLOGY: A total of 121 consecutive inpatient nebulized bronchodilator prescriptions and treatments administered to 93 adult medical inpatients in a university hospital were studied prospectively. Five different aspects of the prescriptions were examined to assess their accuracy and completeness. The administration of each nebulizer treatment was studied using audit of medication charts and interview with ward nurses. RESULTS: No prescription was correct and complete in all five aspects assessed. The most common mistake was failure to state the type and flow rate of driving gas (100%). This was followed by failure to prescribe recommended doses of bronchodilators (46%) and failure to give unambiguous instruction on frequency of treatment (39%). It was found that in only 21.5% of instances was the administration of nebulized bronchodilator drugs optimal. CONCLUSION: This prospective audit has demonstrated major deficiencies in the prescribing and administration of nebulized bronchodilators and it has highlighted the need for a local protocol and continuing staff education. The alternative method of administering bronchodilator via metered dose inhaler with large volume spacer should be evaluated in the treatment of acute airflow obstruction in hospitalized patients. PMID- 12753538 TI - Prevalence and clinical significance of community-acquired penicillin-resistant pneumococcal pneumonia in Thailand. AB - OBJECTIVE: The study aimed to evaluate the prevalence, clinical significance and outcome of community-acquired penicillin-resistant Streptococcus pneumoniae (PRSP) pneumonia in Thailand. METHODOLOGY: We conducted a prospective study in culture-proven pneumococcal pneumonia. Pneumococci were defined as susceptible, intermediate resistance and high resistance according to the definitions of the United States National Committee for Clinical Laboratory Standards (NCCLS). RESULTS: Forty-six consecutive patients were enrolled. Of the S. pneumoniae isolates collected, 41.3% were resistant to penicillin (37% intermediate resistance and 4.3% high resistance). Resistance to other antibiotics was 13% to ceftriaxone (8.7% intermediate resistance and 4.3% high resistance), 34.8% to erythromycin, 39.1% to tetracycline, 26.1% to chloramphenicol, and 43.5% to trimethoprim -sulfamethoxazole. Prior antibiotic use within 3 months was significantly associated with resistance to penicillin. The overall mortality of pneumococcal pneumonia was 26.1%. Multilobar involvement and requirement for mechanical ventilation proved to be associated with mortality. However, resistance to penicillin or ceftriaxone was not associated with death. CONCLUSIONS: According to the US NCCLS guidelines, we found a high prevalence of drug-resistant S. pneumoniae in Thai patients with community-acquired pneumonia. Prior antibiotic use was significantly associated with penicillin resistance. However, the outcome was not related to in vitro penicillin susceptibility of S. pneumoniae isolated from the patients. PMID- 12753539 TI - The effects of air pollution on acute respiratory conditions. AB - OBJECTIVE: Air pollution and its potential health effects are very important worldwide. It is particularly problematic in densely populated cities of developing countries that suffer from a lack of both short- and long-term programmes for air pollution control. We decided to study the short-term effects of air pollution on lung health by assessing the relationship between the levels of six air pollutants and emergency visits for asthma and COPD exacerbations in Tehran, Iran. METHODOLOGY: We monitored the daily attendances for acute respiratory conditions (asthma attacks and COPD exacerbations) to the emergency departments of five major hospitals together with the daily concentrations of six major pollutants during a 5-month period in Tehran. The association between these acute respiratory conditions and the levels of air pollutants was determined by multiple stepwise regression. RESULTS: A correlation was observed between the number of hospital admissions for asthma and the weekly mean concentration of nitrogen dioxide (P < 0.05). The 3-day and 10-day mean concentrations of sulphur dioxide were also found to be directly associated with the number of asthma admissions during this period (P < 0.05). No direct correlation was observed for other variables. CONCLUSION: This study further emphasizes the deleterious effects of air pollution on respiratory health in major populated cities such as Tehran and suggests that increased attention needs to be given to urgent control of air pollution problems. PMID- 12753540 TI - Differences in clinical features between influenza A H1N1, A H3N2, and B in adult patients. AB - OBJECTIVE: The differences in clinical features between influenza A H1N1, A H3N2, and B in the past three influenza seasons were examined. METHODOLOGY: Patients with respiratory symptoms who consulted Kurume University Medical Center, Department of Internal Medicine, Kurume, Fukuoka, Japan, from January to March in 1999, 2000, and 2001 were included. Based on virological and serological findings, the influenza patients were divided into the above three groups for comparison of symptoms and laboratory data. RESULTS: PATIENTS: (n = 196) included 54 with influenza A H1N1, 98 with A H3N2, and 44 with B. Mean ages in the groups were 33 +/- 8.4 years, 41 +/- 15.2 years, and 29 +/- 9.8 years (influenza B patients tended to be younger). Fever was much greater in the A H3N2 group (38.6 +/- 0.46 degrees C) than in the A H1N1 or B groups. This was also true for laboratory indices of viral infection. Gastrointestinal symptoms such as nausea, epigastralgia, and diarrhoea were prominent in influenza B. Myalgia was common in all groups. CONCLUSIONS: Influenza A H3N2 infection was more severe than A H1N1 or B in terms of fever, leukopenia, and C-reactive protein. Myalgia and other symptoms such as fever, headache, general malaise and sore throat were equally frequent in influenza A H3N2, A H1N1, and B infections. Gastrointestinal symptoms were more common in influenza B. PMID- 12753541 TI - Rhinovirus is not detectable in peripheral lung tissue after asthma death. AB - OBJECTIVE: Viral infections are associated with both mild and severe exacerbations of asthma and may therefore be associated with asthma death. As such we hypothesized that it might be possible to detect rhinovirus (RV), the virus most frequently implicated in acute asthma, in lung tissue from patients who died from asthma. METHODOLOGY: We studied archival, wax-embedded lung tissue obtained postmortem from: (i) patients who died from asthma (n = 12), (ii) asthma patients with non-asthma-related death (n = 3), and (iii) non-asthmatic individuals who died from unrelated causes (n = 3). A validated reverse transcription-polymerase chain reaction (RT-PCR) assay was used to detect RV. To confirm RNA preservation, RT-PCR was used to detect expression of the constitutive gene adenine-phosphoribosyl-transferase (APRT). Sensitivity of the assay was assessed using wax-embedded RV-infected cells. RESULTS: Sensitivity of RT-PCR for RV in wax-embedded sections was similar to previous studies (approximately 100 viral copies). Specimens used for study were predominantly of alveolar and small airway origin (< 2 mm). All tissues examined were negative for the presence of RV mRNA and positive for APRT mRNA. CONCLUSIONS: RV infection of the lower airway may be an uncommon cause of fatal asthma. Alternatively, RV may not extend to peripheral airways and more proximal tissue sampling or PCR assays for other viruses may be required to determine an association between viral respiratory tract infection and fatal asthma. PMID- 12753542 TI - Chromosome 9p deletion in squamous metaplasia in cystic lesion of the lung. AB - A 70-year-old man presented with squamous cell carcinoma of the lung. Examination of the resected specimen showed a tumour and a thickly walled cyst not immediately adjacent to the main tumour. The surface of the cyst was lined by squamous metaplastic epithelium. Microsatellite analysis of microdissected specimens was performed with seven markers from four chromosomal regions. In the squamous cell carcinoma, a non-informative homozygosity at two markers on 3p, loss of heterozygosity (LOH) at D5S346 on 5q, and LOH at D9S146, D9S150, and D9S1748 on 9p were detected. In the metaplastic epithelium of the cyst, LOH was detected at D9S1748 on 9p21. This appears to be the first report providing possible evidence of genetic changes by demonstrating allelic loss on 9p21 in the metaplastic epithelium of a cyst. This allelic loss might be related to an early step in carcinogenesis in lung cysts. PMID- 12753543 TI - Microangiopathic haemolytic anaemia and thrombocytopenia following lung volume reduction surgery in a single lung transplant recipient on maintenance tacrolimus (FK506) therapy. AB - Microangiopathic haemolytic anaemia (MAHA) describes intravascular haemolysis due to mechanical destruction of red cells as a result of pathological changes in small blood vessels. It is well recognized as a complication of cyclosporin A therapy in solid organ transplantation but has been uncommonly reported in association with tacrolimus therapy and never before in the setting of lung transplantation. Discussed is a 54-year-old female recipient of a left single lung transplant who developed anaemia, thrombocytopenia and red blood cell fragmentation consistent with MAHA following lung volume reduction surgery (VRS) of the native right lung in the setting of high serum tacrolimus levels. Treatment with fresh frozen plasma and plasmapharesis plus supportive therapy with blood and platelet transfusions resulted in successful resolution of the haemolytic process. Cyclosporin A was substituted for tacrolimus and 18 months later there has been no evidence of recurrence. Tacrolimus therapy is a rare cause of MAHA in solid organ transplants but the diagnosis should be considered if there is an unexplained fall in haemoglobin and/or platelet count in the context of high serum tacrolimus levels. PMID- 12753544 TI - Pulmonary thromboembolism with massive vaginal bleeding due to thrombolytic therapy. AB - Thrombolytic therapy may reduce the morbidity of pulmonary thromboembolism (PTE) and the risk of recurrent disease. The most important complication of thrombolytic therapy is bleeding. We report a patient with PTE complicated by massive vaginal bleeding due to thrombolytic therapy. PMID- 12753545 TI - A case of Legionella pneumophila pneumonia complicated by miliary tuberculosis. AB - A 47-year-old woman was admitted to hospital with severe Legionella pneumonia. The respiratory symptoms improved dramatically and the X-rays revealed a decrease in the diffuse chest infiltrates after treatment with erythromycin and rifampicin. However, chest CT scans showed that the reticulonodular opacities persisted for several weeks after the onset of pneumonia. Two months after admission, the chest X-rays showed the progression of small nodules in both lungs and there was increasing respiratory distress. A diagnosis of miliary tuberculosis was confirmed. The present case should alert physicians to this potentially confusing combination of respiratory pathogens. PMID- 12753547 TI - Testicular measurements and daily sperm output of Tori and Estonian breed stallions. AB - Evaluation of testicular measurements and daily sperm output (DSO) yields valuable information for predicting the reproductive capacity of stallions. The present study evaluated testicular measurements (height, length, width and circumference) and DSO of eight Tori and eight Estonian breed stallions. One ejaculate of semen was collected daily for 10 subsequent days from each stallion. The gel-free volume of semen was measured with a graduated glass cylinder and the sperm concentration was assessed with a Chorjajev chamber. The volume of gel-free fraction was multiplied by the sperm concentration to give the total number of spermatozoa (TSN). The DSO was calculated as mean TSN of collection on days 8-10 in Tori breed stallions and on days 4-10 in Estonian breed stallions. The DSO of Tori breed stallions was 12.9 x 109 spermatozoa and of Estonian breed stallions 4.5 x 109 spermatozoa (p < 0.001). Testicular measurements (in cm) 1 day after the last semen collection were as follows: left testis- height 7.3, length 10.4 and width 7.3 in Tori breed stallions, and 5.9, 8.1 and 5.9, respectively, in Estonian breed stallions; right testis- height 7.4, length 10.6 and width 7.4 in Tori breed stallions, and 5.5, 7.4 and 5.3, respectively, in Estonian breed stallions. All these testicular measurements were significantly smaller in Estonian than in Tori breed stallions (p < 0.001). Testicular circumference was 45.4 and 35.4 cm in Tori and Estonian breed stallions, respectively (p < 0.001). The testicular circumference was correlated with DSO in both Estonian (p < 0.05) and Tori breed stallions (p = 0.071). The results give us valuable information on the reproductive capacity of Tori and Estonian breed stallions. PMID- 12753548 TI - Assessment of progesterone concentration using enzymeimmunoassay, for early pregnancy diagnosis in sheep and goats. AB - The objective of this study was to determine a value of serum progesterone (P4) concentration, assessed using an enzymeimmunoassay (EIA), for the early distinction between pregnant and non-pregnant ewes and goats. Adult, non lactating ewes of Chios (n=53), Berrichon (n=30) and Sfakia (n=45) breeds were synchronized during the breeding season with progestagens and gonadotrophins and mated to fertile rams (Experiment I). Adult, lactating goats of Swiss breeds (Alpine and Saanen, n=104) and indigenous Greek breed (n=45) were synchronized during the transitional season with progestagens, PGF2alpha and gonadotrophins. Cervical artificial insemination (AI) with fresh semen was applied once, 42-44 h after sponge removal (Experiment II). Jugular blood samples were collected on day 19 after sponge removal (ewes) or on day 21 after AI (goats) and serum P4 concentration was determined by EIA. Progesterone concentrations >/=1.0, >/=1.5, >/=2.5 and >/=4.0 ng/ml were tested as indicative of pregnancy. Pregnancy diagnosis was verified on birth. In the case of sheep, using a discriminatory level of 2.5 ng/ml, overall accuracy of pregnancy diagnosis was 91.4% and predictive value of negative and positive diagnoses were 98.3 and 85.3%, respectively. In the case of goats, predictive value of negative diagnosis was 95.8 and 94.0% and predictive value of positive diagnosis 71.3 and 71.7%, for 1.5 and 2.5 ng/ml, respectively; overall accuracy was 79.2% using either level. The other discriminatory levels tested did not improve these results. A significant positive correlation was observed between P4 concentration and the number of lambs or kids born, and further analysis indicated that this relationship is not a simple linear function. Based on the results of this study, P4 concentrations of 2.5 ng/ml in the case of ewes and 1.5-2.5 ng/ml in the case of goats, determined with EIA, are proposed as discriminatory levels between pregnant and non-pregnant animals, at an interval of one oestrous cycle after service. PMID- 12753550 TI - Effects of short-term high carbohydrate or fat intakes on leptin, growth hormone and luteinizing hormone secretions in prepubertal fat-tailed Tuj lambs. AB - The aim of the current study was to evaluate the effects of high carbohydrate or fat diets, fed for 15 days at the end of breeding season, on leptin, GH and LH secretions in prepubertal fat-tailed Tuj lambs. For that purpose, 9-month-old ram lambs were divided into three groups as control group (fed with basal ration, n = 4), high carbohydrate (HC) group, basal ration plus barley, n = 4), or high fat (HF) group (basal ration plus by-pass fat, n = 4). For the measurement of leptin and GH, blood plasma samples were collected on days 1, 4, 9 and 14 of the experiment. For the measurement of LH pulse frequency, serial blood samples were collected every 15 min for 6 h on day 14. Lambs were weighed and body condition scored (BCS) on days 1 and 15. Body weight and BCS increased towards the end of the study (p < 0.05). The BCS was higher in high energy groups at the end of the experiment (p < 0.05). Diet affected plasma leptin concentrations (p = 0.002) but time did not. The GH concentrations were not affected by diet or time. The LH pulse frequency appeared to be higher in HC and HF groups but there were no statistical difference between the groups. There was a significant positive relationship between overall BCS and corresponding leptin concentrations (R2 = 0.263; p = 0.010) and between LH pulse frequency and leptin concentrations (R2 = 0.594; p = 0.003). In conclusion, the present study suggests that rather than type of energy, amount of energy intake and body energy reserves are much important regulators of plasma leptin concentrations and LH pulse frequency in fat-tailed Tuj lambs. PMID- 12753549 TI - Expression of osteopontin (OPN) mRNA in bovine ovarian follicles and corpora lutea. AB - The matricellular protein osteopontin (OPN) plays a role in various physiological processes, including angiogenesis and tissue remodelling. As these processes are essential for the maintenance of ovarian physiology, the aim of the study was to investigate the expression of OPN (mRNA) in ovarian cells and to evaluate whether it can be regulated by gonadotrophins. Using conventional RT-PCR and real-time PCR, we have detected and quantified OPN mRNA as well as glyceraldehyde-3 phosphate dehydrogenase (GAPDH) mRNA expression in bovine granulosa, theca and luteal cells. In all cells examined, both genes were found in equal amounts and no striking variations in the expression could be observed between granulosa, theca and luteal cells. Furthermore, no effect on either OPN or GAPDH mRNA expression was evident after culturing ovarian cells in the presence of gonadotrophic hormones, although the cells were still highly responsive in terms of cAMP formation. Although neither variations between different cell types nor a regulation of OPN mRNA expression by gonadotrophic hormones could be detected, the high and unambiguous mRNA expression in steroidogenic cells suggests that OPN should be added to the growing list of intraovarian factors which may be involved in ovarian physiology. PMID- 12753551 TI - The effects of the periodical use of in-feed chlortetracycline on the reproductive performance of gilts and sows of a commercial pig farm with a history of clinical and subclinical viral and bacterial infections. AB - The objective of this study was to assess the effects of in-feed chlortetracycline (CTC) as a measure of preventing or minimizing infectious problems of reproductive failure in gilts and sows. In a farm of 400 Large White x Landrace gilts and sows with a clinical history of porcine respiratory and reproductive syndrome (PRRS) virus, the animals were treated with CTC. Treatment consisted of 10 g CTC sow/day for 15 days every 3 months. It improved the health status of sows by decreasing post-farrowing clinical mastitis and vaginal discharges, abortions, return-to-oestrus and irregular return-to-oestrus rates. These beneficial effects had a positive impact on the performance of the litter. More piglets were born live and weaned. These positive effects improved with repeated use of CTC. The serological evidence of PRRS virus, Leptospira spp. and Chlamydia spp. and the subsequent beneficial use of the antimicrobial agent indicate that reproductive failure, possibly resulting from the bacterial agents can be controlled with in-feed use of broad spectrum antimicrobials. PMID- 12753552 TI - Postnatal hypothyroidism does not affect prepubertal testis development in boars. AB - Young boars were treated with propiothiouracil to induce hypothyroidism to examine its effects on postnatal testicular development. Treatments with 0.1% 4 propyl-2-thiouracil (PTU) in drinking water started after weaning, at 3 weeks of age and all boars were severely hypothyroid at 6 weeks of age as determined by measuring T3 and T4 in blood plasma. Boars were castrated at different ages up to 20 weeks and their testes used for histological and immunohistochemical analyses. Although small but significant reduction in testis weight was observed from 8 to 12 weeks of age, this was not accompanied by significant difference in testicular volume. By 20 weeks of age, at the beginning of puberty, the differences in testis weights between control and treated groups of boars disappeared suggesting there is no lasting effect of hypothyroidism on postnatal development of boar testis. Immunohistochemical staining was used to determine the presence of molecular markers in both Sertoli and Leydig cells. Again, there were no differences between testes from control and treated boars in the pattern or intensity of immunostaining using antibodies against 3beta-hydroxysteroid dehydrogenase, antimullerian hormone or proliferating cell nuclear antigen (PCNA). Immunostaining with antibodies against PCNA showed interesting results as it was observed that Sertoli cells still express this marker of proliferating cells at 14 weeks of age, later than previously suggested cessation of Sertoli cell proliferation. This study suggests that hypothyroidism in boars does not have similar effects on postnatal testis development as reported in some other species. PMID- 12753553 TI - Conception rates after AI in Swedish red and white dairy heifers: relationship with progesterone concentrations at AI. AB - Blood samples were collected from 211 dairy heifers at the time of field insemination [artificial insemination (AI)]. Heifers were defined as either first service heifers (n = 91) or third to eigth-service heifers [presumed (third AI, n = 60)] or well-defined repeat breeders (greater than the fourth AI, n = 60). Plasma progesterone concentrations at AI were evaluated in relation to oestrous behaviour at AI and conception rates post-AI. Conception rates after third AI were good, but conception rates decreased markedly after fourth AI and onwards. Those heifers that did not become pregnant after AI had significantly higher basal progesterone concentrations (>0.5 nmol/l) at AI, so-called suprabasal concentrations, compared with those which conceived after AI (irrespective of the number of AI practised). Relative risk for repeat breeding was 58% after AI performed at suprabasal progesterone concentrations, while it was 42% at basal (0.5) progesterone concentrations. Results from this field study confirm results obtained in earlier controlled studies of repeat-breeder heifers (RBH), indicating that the current definition of repeat breeding should be retained. Analysis of suprabasal progesterone concentrations could be considered as a tool for identification of RBH, provided that heat detection and AI timing are optimal. Besides considering the direct costs involved in repeat breeding, it remains to be determined whether it would be economically beneficial to identify and exclude RBH from the breeding population. PMID- 12753554 TI - Manipulated mouse embryos as bioassay system for water quality control. AB - Mouse pronuclear stage embryos with intact slit zona pellucida (manipulated) were cultured in vitro until the hatched blastocyst stage in simplex optimized medium with higher K+ concentration (KSOM) prepared with three different water types: tap, deionized reverse osmosis (D-O) water and Milli-Q system (M-Q) water. The culture media were supplemented with or without protein and ethylenediaminetetraacetic acid (EDTA, disodium salt). The rates of hatched blastocysts were significantly affected (p < 0.01) by micromanipulation, protein supplement and water source. The water source has no influence (p > 0.05) on development in EDTA-supplemented protein-free culture media, whereas in EDTA- and protein-free culture media, the water quality significantly (p < 0.001) affected the rates of development, with higher rates in media prepared with M-Q water. The micromanipulated embryos showed higher sensitivity to the water quality (p < 0.01). It worth mentioning that the rates of hatched blastocysts in protein-free culture media were very low (0-7.5%). Furthermore, the three different water types were analysed by measuring the electrical conductivity, inorganic ions, total organic carbon and endotoxins to evaluate the purity. M-Q water showed the lowest levels of inorganic ion, total organic carbon and endotoxin concentrations. We concluded that manipulated mouse embryos are good system to evaluate the quality of water used in biological system. PMID- 12753555 TI - Apoptosis of granulosa cells: a review on the role of MAPK-signalling modules. AB - Recent studies suggest that ovarian follicular atresia is associated with DNA fragmentation and degeneration of granulosa cells, the hallmark of programmed cell death or apoptosis. Apoptosis of granulosa cells play a major role in follicular atresia. These studies have also demonstrated the involvement of tumour suppressors, apoptotic proteins and survival factors. These factors contribute to the developmental decision as to whether the ovarian follicles mature or undergo atresia. However, the precise temporal and molecular events involved in the apoptotic pathways in this process need to be elucidated. The present report summarizes the role of Jun N-terminal kinase (JNK), p38 mitogen activated protein kinase (p38 MAPK), and extracellular-signal regulated kinase (ERK)-signalling module in the regulation of pro- and anti-apoptotic factors of the granulosa cells in regulating follicular atresia. The findings presented here suggest that the loss of tropic hormone support is translated into the attenuation of Raf-1-MAPK/ERK kinase (MEK)-ERK-signalling pathway of the granulosa cells and this results in the decreased phosphorylation of the pro apoptotic BAD. PMID- 12753556 TI - Ovarian follicular dynamics in buffalo cows (Bubalus bubalis). AB - Follicular growth in Egyptian buffalo cows was monitored using genital tracts from 200 buffalo cows collected immediately after slaughter. According to the morphological appearance of the corpus luteum (CL), the corresponding oestrous cycle was divided into four stages: A (days 1-4), B (days 5-10), C (days 11-17) and D (days 18-21). Within these stages the follicular population on the ovaries was evaluated and the dominant follicle (DF) determined in all recovered ovaries. The functional status of the DF and the largest sub-dominant follicles was examined by histological examination in 31 cases, and Radio Immunoassay (RIA) analyses for estradiol-17beta (E2) and progesterone (P4) was performed in the follicular fluid in 23 of the DF. The results showed that DFs changed their endocrine character within the stages of the oestrous cycle. The DFs between days 5 and 10 were functionally active (E2-dominant; non-atretic) in most of the cases. Between days 11 and day 17 half of the DFs became functionally inactive (P4-dominant; atretic). At days 18-21 all of the DF became functionally active and non-atretic. In the specimens that carried two large follicles one of them was regularly atretic and P4-dominant whereas the other was non-atretic and E2 dominant. Between days 18 and 21 all ovaries examined showed at least one large follicle. These findings suggest that in most of the cases follicular dynamics occurs in two wave-like patterns in the Egyptian buffalo cows. PMID- 12753557 TI - Secretory patterns of prolactin in dogs: circannual and ultradian rhythms. AB - The objectives of the present study were to characterize in dogs circannual and ultradian prolactin (PRL) secretory patterns and also to compare gender differences in the ultradian period of study in the Southern hemisphere. Blood samples were collected at 15-min intervals for 2.5 h from seven male and seven female dogs and a single monthly sampling, over a 1-year time span, from six male dogs for the ultradian and circannual studies, respectively. Plasma PRL was measured by a homologous enzyme immunometric assay. The ultradian study evidenced PRL elevations suggesting pulsatile secretion in both genders. Significantly higher mean smoothed baseline (ng / ml [7.02 +/- 1.2 vs 1.23 +/- 1.0, p < 0.01]) and AUC (ng/ml * 2.5 h [25.2 +/- 3.8 vs 4.4 +/- 3.8, p < 0.01]) were found in females when compared with males. In the circannual study, plasma PRL concentrations did not statistically differ among the months of the year. When grouped together the 3 months with a longer daylight had significantly higher PRL concentrations than the 3 months with the shortest (2.31 +/- 0.37 vs 0.96 +/- 0.37, p < 0.01). The correlation between length of daylight and PRL concentrations was 0.24, p < 0.05. It is concluded that PRL does have a circannual rhythmicity and that there are ultradian gender-related differences in the period under study in these groups of dogs. This study also demonstrates plasma PRL elevations suggesting pulsatile secretion in male dogs. PMID- 12753558 TI - Sperm pre-incubation prior to insemination affects the sex ratio of bovine embryos produced in vitro. AB - The objective of the present study was to determine whether sperm incubation prior to oocyte insemination in vitro affects the sex ratio of resulting blastocyst. Cumulus-oocyte-complexes (COCs) collected from slaughterhouse ovaries were matured in vitro and inseminated with frozen-thawed semen of three proven artificial insemination (AI) bulls pre-incubated in vitro in Sperm-Talp for 6 and 24 h. On day-9 blastocysts were collected and processed for sex determination. More than 80% of blastocyst were successfully sexed. There were no significant differences in cleavage and blastocyst rates using sperm pre-incubated for 6 h as compared with the 0-h pre-incubation control group. The cleavage and blastocyst rates were significantly lower in the 24-h pre-incubation group. The male to female ratio, when compared with the theoretical 1 : 1, differed significantly in favour of females among hatched (viable) blastocysts derived from sperm pre incubated for 24 h prior to insemination as well as among all blastocytsts in the 6-h group. Moreover, when the sperm treatment was considered, the sex ratio was affected only among hatched blastocysts in 24-h pre-incubation group. It was concluded that prolonged sperm pre-incubation influences the rate of development and the sex ratio among hatched blastocysts. PMID- 12753559 TI - Effect of progesterone prior to GnRH-PGF2alpha treatment on induction of oestrus and pregnancy in anoestrous Awassi ewes. AB - An experiment was conducted to examine the effect of progesterone prior to a GnRH PGF2alpha treatment on oestrus and pregnancy in seasonally anoestrous Awassi ewes. Twenty-four ewes were randomly assigned to three groups to be pre-treated with 60 mg medroxyprogesterone acetate sponges (group A), 600 mg progesterone sponges (group B) or blank sponges (group C) for 4 days. All ewes were injected with 100 microg of GnRH 24 h after sponge removal followed, 5 days later, by 20 mg PGF2alpha injection. Ewes were exposed to three fertile rams at the time of PGF2alpha injection (day 0, 0 h) and were checked for breeding marks at 6-h intervals for 5 days. Blood samples were collected from all ewes 1 day (day -10) prior to sponge insertion, at the time of sponge removal (day -6), 1 day following sponge removal (day -5, at the time of GnRH injection) and at the time of PGF2alpha injection (day 0) for analysis of progesterone. Progesterone concentrations on days -10 and -5 were basal and averaged 0.2 +/- 0.04 and 0.2 +/ 0.2 ng/ml, respectively. Progesterone concentrations on day -6 were elevated only in group B ewes and were higher (p < 0.0001) than those of groups A and C. Progesterone concentrations on day 0 were higher (p = 0.002) in groups A and B than group C. Oestrous responses occurred only in ewes of groups A and B (p > 0.05). Induced oestrus conception rate was greater (p < 0.01) in group A than groups B and C. Ewes returned to oestrus 17-20 days following day 0 were two of eight, six of eight and three of eight of groups A, B and C, respectively, all of which eventually lambed. The overall lambing rate was 82% in progesterone-primed ewes compared with only 38% non-progesterone-primed ewes (p < 0.05). Progesterone priming apparently sensitizes GnRH-PGF2alpha-treated seasonally anoestrous ewes and increases their response in oestrus and pregnancy rates. PMID- 12753560 TI - 24-hour secretion patterns of plasma oestradiol 17beta in pony mares in late gestation. AB - The mare exhibits nocturnal uterine contractions in the last 6 days of gestation. It is hypothesized that estradiol 17beta (O17beta) may be associated with the nightly increase in uterine contractions. The 24-h secretion pattern of plasma O17beta was measured in 3 pony mares in late gestation to identify changes in release as the mare neared parturition. Blood was collected weekly at 08:00 hours beginning on day 240 and every third day from day 330 until delivery. Serial blood samples were collected from each mare every 30-min for 24-h beginning on gestation day 310 and every sixth day thereafter until parturition. Concentrations of O17beta were elevated at night with lowest concentrations occurring directly before sunset (p < 0.01). The natural log of the variance was increased at sunset (p < 0.01) and was decreased during the 6-h period immediately after sunrise. This pattern was especially evident in the 6 days that preceded parturition. The contrast between nocturnal and daytime concentrations of O17beta in the last 6 days of gestation may contribute to night-time delivery in the mare. PMID- 12753561 TI - Effect of modified eros centre on some reproductive parameters of the sow. AB - The effect of a modified eros centre on weaning to oestrus interval, follicle size, ovulation and farrowing rate and total born litter size was investigated. In modified eros centre 94.4% and in group housing 79.1% of the sows (p < 0.01) expressed oestrus within 10 days post-weaning. Weaning to oestrus interval was shorter (p < 0.001) for sows kept in modified eros centre. The interval from onset of oestrus to the time of ovulation was longer for sows in group housing (p=0.05). The time of ovulation was negatively correlated (r=-0.50) with the interval from weaning to oestrus (p=0.005). The time of ovulation after onset of oestrus was significantly (p < 0.05) shorter for sows expressing oestrus within 2 4 days of weaning, compared with the animals that expressed oestrus between days 5 and 6 post-weaning and was shortest for sows expressing oestrus after day 6 post-weaning. Farrowing rate was not affected by a modified eros centre. Litter size tended to be smaller in group-housed weaned sows (p=0.10). The timing of last artificial insemination relative to time of ovulation did not affect litter size (p > 0.10). The implication of these results is that a modified eros centre may improve some of the post-weaning oestrous parameters of the sow. PMID- 12753562 TI - Biological predictors of lithium response in bipolar disorder. AB - In order to prescribe lithium appropriately to patients with bipolar disorder, predictors of lithium response are helpful. The present paper reviews the biological predictors of lithium response. As a positive predictor of lithium response, the following have been reported: strong loudness dependence of the auditory-evoked N1/P2-response; higher brain lithium concentration; lower inositol monophosphatase (IMPase) mRNA expression; higher serotonin-induced calcium mobilization; increased N-acetyl-aspartate peak and decreased myo inositol peak; white matter hyperintensity; decreased intracellular pH; higher frequency of phospholipase C gamma-1 (PLCG1)-5 repeat and PLCG1-8 repeat; and C973A polymorphism in the inositol polyphosphate 1-phosphatase gene. In contrast the following have been reported as a predictor of negative lithium response: epileptiform abnormality of electroencephalography; human leukocyte antigen type A3; decreased phosphocreatine peak area after photic stimulation; and homozygotes for the short variant of the serotonin transporter gene. Most of the possible biological predictors of better lithium response, such as lower IMPase mRNA levels, white matter hyperintensity, lower brain intracellular pH, enhanced calcium response, and PLCG1-5 repeat had been detected as risk factors for bipolar disorder, suggesting that bipolar disorder responding well to maintenance lithium treatment is a distinct category having a certain neurobiological basis, although these findings need further replication. The search for biological predictors of lithium response is still in its infancy. Most of the laboratory or neuroimaging techniques used in these studies are not easily performed in clinical settings, so the development of an easy and useful laboratory test is needed. PMID- 12753563 TI - Influence of depressed mood on neuropsychologic performance in HIV-seropositive drug users. AB - Some studies point out that depression affects the performance of HIV patients in neuropsychological tasks, but at present this effect is not clear. The purpose of the present paper was to study whether the presence of symptoms of depression affects the neuropsychologic performance of seropositive drug users in tasks of attention/concentration, learning and memory, language, construction and visuospatial function, speed of motor performance, cognitive flexibility, manual skill and concept formation and reasoning. In order to carry out this research a sample consisting of 127 male volunteer subjects was used. These subjects were distributed in four groups: one group consisted of HIV-seropositive drug users with symptoms of depression (n = 33); the second group consisted of HIV seropositive drug users without symptoms of depression (n = 47); the third group was formed by HIV-seronegative drug users with symptoms of depression (n = 15) and the fourth group was formed by HIV-seronegative drug users without symptoms of depression (n = 32). The results reveal the effect of symptoms of depression (evaluated by the Beck Depression Inventory) on the neuropsychologic performance of seropositive drug users. This effect, however, was not observed in the seronegative group. These findings lead us to suggest that symptoms of depression constitute a risk factor for presenting neuropsychologic disturbances in seropositive subjects, which could well be acting as a factor that foments the neuropsychological effects of HIV. PMID- 12753564 TI - Discrepancy between subjective and objective sleep in patients with depression. AB - The literature investigating the relationship between objective and subjective sleep in depressed patients is limited and the results are inconsistent. Furthermore, many factors that influence the aforementioned relationship have not been investigated. The present study was carried out to clarify the characteristics of self-estimation of sleep in depressed patients. Sleep was estimated concurrently using a sleep log and polysomnography for 5 consecutive days to investigate the relationship between subjective sleep estimation and objective sleep estimation in 23 patients with major depression (Diagnostic and Statistical Manual of Mental Disorders, 3rd edn, revised; DSM-III-R). Factors related to a discrepancy between both types of estimation were identified. The subjective total sleep time showed a significant, but moderate, positive correlation (correlation coefficient: 0.63) with the objective total sleep time. The degree of discrepancy was significantly correlated with various objective sleep variables and severity of depression. In the underestimation group in which the subjective total sleep time was shorter than the objective total sleep time, the objective total sleep time and slow-wave sleep time were shorter, age was greater and the extroversion score (Maudsley Personality Inventory) was lower than in the overestimation group in which the subjective total sleep time was longer than the objective total sleep time. The data suggest that subjective sleep estimation in depressed patients is influenced by their objective sleep, severity of depression, age and personality. PMID- 12753565 TI - Sleep disturbances and depression in the elderly in Japan. AB - The purpose of the present study was to evaluate the relationship between sleep disturbances and depression in the Japanese elderly. METHODS: These investigations in the Japanese elderly were carried out with the Geriatric Depression Scale, the Pittsburgh Sleep Quality Index, and questions on restless legs syndrome and nocturnal eating disorder. A total of 2023 people (male: 1008; female: 1015; average age: 74.2 +/- 6.3 years) were analyzed by chi2 test and simple and multiple logistic regression. The prevalence of sleep disturbance was 37.3% and that of depression was 31.3%. Female gender and/or older (> or =75 years) age were significantly associated with depression. Characteristics in depressive elderly were poor sleep efficiency, sleep disturbances due to difficulty of initiating sleep (DIS), breathing discomfort, coldness and pain, poor subjective sleep quality and lack of enthusiasm for activities. Sleep disturbances due to using the bathroom, breathing discomfort and coldness and long sleep latency were associated with depression in younger (65-74 years) men. Sleep disturbance due to DIS was associated with depression in older (> or =75 years) men. Sleep disturbance due to pain was associated with depression in younger and older women. Poor sleep efficiency was associated with depression in older women. Poor subjective sleep quality was associated with depression in younger and older men and younger women. Lack of enthusiasm was associated with depression in younger and older men and older women. Restless legs syndrome was statistically significantly associated with depression in younger men. It is concluded that sleep disturbance and depression among the Japanese elderly are closely related symptoms. The features of sleep disturbance with depression differed with sex and age. PMID- 12753566 TI - Prenatal underdevelopment and schizophrenia: a case report of monozygotic twins. AB - A case is described herein of monozygotic twins discordant for schizophrenia that illustrates possible causal events of prenatal underdevelopment, morphological changes in the brain, poor premorbid functioning, and the development of schizophrenia. The affected twin was born with a birthweight of 1620 g, whereas the unaffected twin weighed 2300 g at birth. Marked differences in sociability and intelligence were observed between the twins from early childhood. Magnetic resonance imaging of the brain revealed high-intensity signals in the white matter and enlarged ventricles in the affected twin, while no such abnormality was detected in the well twin. This twin pair suggests a possibility that hypoxic brain damage associated with prenatal underdevelopment may be a causal factor for abnormalities in psychosocial development and subsequent schizophrenia. PMID- 12753567 TI - Behavioral parent training for Taiwanese parents of children with attention deficit/hyperactivity disorder. AB - It has been observed that it is relatively difficult for children with attention deficit/hyperactivity disorder (ADHD) to follow social rules and behave in a socially desirable manner. The ADHD children in Chinese culture, which emphasizes Confucian values, might encounter even greater adjustment difficulties. The purpose of the present study is to implement a behavioral parent training program in a Confucian environment and examine its effectiveness. Twenty-three ADHD preschoolers (age: 3-6 years) and their parents were selected to participate in the present study. Fourteen of these 23 parents completed a 10-session parent training program. Parent ratings of ADHD/oppositional defiant disorder (ODD) symptoms and problem behaviors at home were collected at the first, fourth, sixth, seventh, and tenth sessions. Three instruments were used to evaluate treatment outcome: the Disruptive Behavior Rating Scale-Parent Form, Child Attention Profile, and Home Situations Questionnaire. The results showed that both ADHD/ODD symptoms and home behaviors of these 14 children improved significantly after the parent training. There was also a significant decline in the severity of symptoms and problem behaviors at home with the progression of training. These findings support the effectiveness of this parent training program for parents of ADHD children in an environment of Confucianism. Limitations of the present study and future direction for research are discussed. PMID- 12753569 TI - Differences in emotional distress between breast tumor patients with emotional inhibition and those with emotional expression. AB - The differences in affective status between patients who restrain their negative emotion and those who express negative emotion after being given their breast cancer diagnosis were studied using the Profile of Mood States (POMS) at two sessions: (i) at the first visit to the outpatient surgery clinic, and (ii) immediately after being given the diagnosis of breast cancer. Eighty-seven patients completed the POMS and the Courtauld Emotional Control Scale (CECS) at the first visit to the outpatient surgery clinic at Shiga University of Medical Science Hospital. They also completed the POMS immediately after being given the diagnosis of breast cancer. Breast cancer patients who restrain their negative emotion (n = 8) were highly anxious, depressed and confused after being given the diagnosis compared to breast cancer patients who express negative emotion (n = 8). Emotional distress in benign breast tumor patients was reduced after being given the diagnosis regardless of the trend of emotional inhibition. That is, emotional distress in patients who restrain their emotions was considerably increased compared with that of patients who expressed their emotions when they were faced with a life-threatening disease. These results suggest that it may be therapeutic to advise breast cancer patients to express their negative emotion. PMID- 12753568 TI - Polymorphisms of interleukin-4 promoter and receptor gene for schizophrenia in the Korean population. AB - The purpose of the present paper was to investigate the interleukin (IL)-4 promoter gene -590 and receptor alpha (Ralpha) gene 1902 polymorphism in Korean schizophrenic patients. A total of 222 Korean patients diagnosed with schizophrenia according to Diagnostic and Statistical Manual of Mental Disorders (4th edn; DSM-IV) and 165 normal healthy controls participated in the present study. The DNA was extracted from whole blood using proteinase K, and the IL-4 promoter and receptor gene were amplified by polymerase chain reaction. The genotype was determined using single-strand conformation polymorphism (SSCP) analysis. The distribution of the alleles and genotypes in patients with schizophrenia was not significantly different from those of controls. In conclusion, these results suggest that the polymorphisms in IL-4 promoter gene 590 and IL-4 Ralpha gene 1902 are not involved in the pathophysiology of schizophrenia in the Korean population. PMID- 12753570 TI - Efficacy of open-system social skills training in inpatients with mood, neurotic and eating disorders. AB - Open-system social skills training (SST) was performed in an open psychiatric ward of the hospital of Yokohama City University. Between June 1998 and March 2000, 223 patients were being treated for various mental disorders and 136 of these patients voluntarily participated in the open-system SST at least once. The SST participants' ages were 37.2 +/- 16.9 years and the admission period was 102.6 +/- 61.4 days, while non-participants' ages were 49.8 +/- 18.8 years and the admission period was 71.8 +/- 55.6 days. The correlation between participation time and the admission period showed a ratio of approximately 0.5. As for diagnoses, schizophrenia, eating disorder and personality disorder patients tended to participate in SST, while organic mental disorder patients tended to be non-participants. After October 1998 there were 26 patients continuing to attend SST who participated in the evaluation study that compared social skills estimation before SST with that after SST by self-evaluation and by the staff using a social skills questionnaire. After SST sessions, the average staff evaluation score was 43.9% in schizophrenia, 64.4% in mood disorder, 64.9% in neurotic disorder and 55.3% in eating disorders, while they were 29.1%, 33.8%, 44.4% and 34.5% before the sessions, respectively. After the SST sessions, mood disorder patients showed a 25-30% increase of both self-estimation and staff estimation in all subcategories of social skills. These findings suggest that SST was effective for all patients motivated to improve their social skills despite diagnoses and that the SST program had different effects in each diagnosis group. PMID- 12753571 TI - Comparison of hangover effects among triazolam, flunitrazepam and quazepam in healthy subjects: a preliminary report. AB - The aim of the present study was to compare the hangover effects of night-time administration of triazolam (0.25 mg), flunitrazepam (1 mg) and quazepam (15 mg) in healthy subjects. Daytime sleepiness and performance level following the night time administration of the drugs were assessed using Standford Sleepiness Scale (SSS), Sleep Evaluation Questionnaire (SEQ), Multiple Sleep Latency Test (MSLT), actigraphy recordings and Continuous Performance Test (CPT). Fifteen healthy volunteers were given one of the three hypnotics at each drug session, which lasted for 1 week, in a single-blind cross-over fashion. No significant between drug difference was observed for the psychomotor performance assessed by CPT. Subjective hangover effects assessed by SSS and SEQ in the morning were prominent for flunitrazepam and quazepam relative to triazolam, whereas objective indices such as MSLT or activity counts obtained in actigraphy indicated a marked hangover effect of quazepam compared with the other two compounds restrictively in the afternoon, which were nearly in accordance with their pharmacokinetic profiles. PMID- 12753572 TI - Bone marrow transplantation in subjects with mental disorders. AB - Bone marrow transplantation (BMT) is a critical treatment of malignant illnesses including leukemia and others. Successful achievement of BMT requires the patients to tolerate isolation for several weeks to avoid infections. They are also required to follow several regulations and instructions to survive the treatment because the patients' physical condition is complicated due to the malignant illness, preparatory treatment and transplant of bone marrow from other subjects. These could be a significant challenge for patients with mental disorders. Here the cases are reported of seven leukemia patients who were referred to the Metropolitan Komagome Hospital for BMT from April 1996 through May 2000, who had been suffering from mental disorders, including schizophrenia, bipolar I mood disorder, panic disorder, dysthymic disorder, autistic disorder, and borderline personality disorder, prior to the treatment. The BMT was achieved in six out of the seven subjects; the exception was a subject with borderline personality disorder. Psychiatric treatments, including medication, to improve and maintain mental status appeared to be critical for the achievement of BMT in several patients. Understanding of the status of the malignant disease and the role of BMT was another significant issue. Test admission seemed to be helpful to reduce concerns and anxiety both in the patients and hospital staff. PMID- 12753573 TI - Effective open-label treatment of premenstrual dysphoric disorder with venlafaxine. AB - Various studies have demonstrated the efficacy of selective serotonergic re uptake inhibitors in the treatment of premenstrual dysphoric disorder (PMDD). But the effectiveness of novel antidepressant, venlafaxine, in PMDD has been reported in only one Western study. The purpose of the present open-label study was to provide preliminary data on the effectiveness of venlafaxine for Asian women with PMDD. Thirty women with PMDD were enrolled and treated with a flexible dosage of venlafaxine for two menstrual cycles. Responses were assessed every 2 weeks. Outcome measures included the scores of the Prospective Record of the Impact and Severity of Menstrual Symptomatology (PRISM) calendar, self-rating Zung Depressive Scale (Zung), State and Trait Anxiety Inventory (STAI), Hamilton Rating Scale for Depression/Anxiety (HAM-D/HAM-A), and the Clinical Global Impression scale (CGI). Twenty patients completed the trial. All patients had significant improvement of the mood and behavior components in the PRISM calendar. The effects of active treatment were marked by the first active cycle of menstruation. Venlafaxine at a mean dose of 60.1 +/- 29.1 mg per day was effective in reducing PMDD symptoms. The results of the present open trial indicated that venlafaxine is effective in the treatment of ethnic Taiwanese women with PMDD. PMID- 12753574 TI - Understanding personality traits from early life experiences. AB - The contribution of early experiences towards the onset of personality disorder has often been stressed. However, the contribution to trait personality has received less attention. To examine the impact of early experiences on the development of personality, two subscale scores of the Eysenck Personality Questionnaire (EPQ): neuroticism (N) and extroversion (E), were used to assess a total of 220 residents of a rural city of Japan (aged > or =18 years). After controlling for age and social desirability response bias, the N score of men could be predicted by the experience of relocation; the E score of men by high parental care and low parental overprotection; and the E score of women by the experience of death of a sibling. Personality traits in a non-patient population may be explained by early experiences. PMID- 12753575 TI - Phenomenology of hallucinations: a factor analytic approach. AB - The present study was carried out with the aim of obtaining a factor analytic solution of parameters of hallucinations in schizophrenia. Seventy-five chronic hallucinating schizophrenic patients were assessed on the Phenomenology of Hallucinations Scale and Brief Psychiatric Rating Scale. A factor analytic solution was obtained by principal component analysis using varimax rotation. Two factors, 'reality of hallucinatory perception' and 'immersion in hallucination', were obtained. Findings are discussed in relation to existing literature. PMID- 12753576 TI - Regional cerebral blood flow in delirium patients. AB - The purpose of the present paper was to determine the possible mechanism of delirium by using xenon-enhanced computed tomography to measure the regional cerebral blood flow (rCBF) of the patients both during delirium and after improvement from delirium. The rCBF measurements of the frontal, temporal and occipital cortex during delirium ranged from 31.4 to 39.6 mL/100 g per min; the rCBF of the thalamus and basal ganglia ranged from 47.5 to 52.4 mL/100 g per min. After recovery from delirium the rCBF of both areas returned to normal. The findings that reduced rCBF during delirium becomes normal once delirium improves suggest that a possible cause of delirium may be the cerebral hypoperfusion. PMID- 12753577 TI - Eating disorder with hyperthyroidism. PMID- 12753578 TI - Plasma nitrate and nitrite levels of patients with eating disorders. PMID- 12753579 TI - Kami-shoyo-san is usually used for women. PMID- 12753580 TI - The sfr6 mutant of Arabidopsis is defective in transcriptional activation via CBF/DREB1 and DREB2 and shows sensitivity to osmotic stress. AB - The sfr6 mutant of Arabidopsis displays a deficit in freezing tolerance after cold acclimation. We previously observed that the transcripts of three cold-, ABA and drought-inducible genes, each having a C-repeat motif or the drought responsive element (CRT/DRE) in its promoter, failed to normally accumulate in this mutant. We now report that the effects of sfr6 upon transcript levels are reflected in the levels of the encoded proteins, confirming that the cold inducible protein expression is affected by the sfr6 mutation. Using microarray analysis, we found not only that this effect may be general to cold-inducible genes with CRT/DRE promoter elements, but also that it extends to some other genes whose promoters lack a CRT/DRE element. The role of the CRT/DRE has been empirically tested by use of a synthetic promoter, confirming that the CRT/DRE is sufficient to confer the sfr6 effect upon expression. Tolerance of osmotic stress was also found to be reduced in sfr6, consistent with a role in osmotic stress tolerance for the cold-, ABA- and drought-inducible genes whose expression is affected by the sfr6 mutation. PMID- 12753581 TI - Genetic characterization of a pentatricopeptide repeat protein gene, orf687, that restores fertility in the cytoplasmic male-sterile Kosena radish. AB - Cytoplasmic male sterility (CMS) in plants is a maternally inherited inability to produce functional pollen, and is often associated with mitochondrial DNA abnormalities. Specific nuclear loci that suppress CMS, termed as restorers of fertility (Rf), have been identified. Previously, we identified an Rf for the CMS Kosena radish and used genetic analysis to identify the locus and create a contig covering the critical interval. To identify the Rf gene, we introduced each of the lambda and cosmid clones into the CMS Brassica napus and scored for fertility restoration. Fertility restoration was observed when one of the lambda clones was introduced into the CMS B. napus. Furthermore, introduction of a 4.7-kb BamHI/HpaI fragment of the lambda clone is enough to restore male fertility. A cDNA strand isolated from a positive fragment contained a predicted protein (ORF687) of 687 amino acids comprising 16 repeats of the 35-amino acid pentatricopeptide repeat (PPR) motif. Kosena CMS radish plants were found to express an allele of this gene possessing four substituted amino acids in the second and third repeats of the PPR suggesting that the domains formed by these repeats in ORF687 are essential for fertility restoration. Protein levels of the Kosena CMS-associated mitochondrial protein ORF125 were considerably reduced in plants in which fertility was restored, although mRNA expression was normal. Regarding the possible role for PPR-containing proteins in the regulation of the mitochondrial gene, we propose that ORF687 functions either directly or indirectly to lower the levels of ORF125, resulting in the restoration of fertility in CMS plants. PMID- 12753582 TI - Cell cycle function of a rice B2-type cyclin interacting with a B-type cyclin dependent kinase. AB - Cyclin-dependent kinases (CDKs) are involved in the control of cell cycle progression. Plant A-type CDKs are functional homologs of yeast Cdc2/Cdc28 and are expressed throughout the cell cycle. In contrast, B-type CDK (CDKB) is a family of mitotic CDKs expressed during the S/M phase, and its precise function remains unknown. Here, we identified two B2-type cyclins, CycB2;1 and CycB2;2, as a specific partner of rice CDKB2;1. The CDKB2;1-CycB2 complexes produced in insect cells showed a significant level of kinase activity in vitro, suggesting that CycB2 binds to and activates CDKB2. We then expressed green fluorescent protein (GFP)-fused CDKB2;1 and CycB2;2 in tobacco BY2 cells to investigate their subcellular localization during mitosis. Surprisingly, the fluorescence signal of CDKB2;1-GFP was tightly associated with chromosome alignment as well as with spindle structure during the metaphase. During the telophase, the signal was localized to the spindle midzone and the separating sister chromosomes, and then to the phragmoplast. On the other hand, the CycB2;2-GFP fluorescence signal was detected in nuclei during the interphase and prophase, moved to the metaphase chromosomes, and then disappeared completely after the cells passed through the metaphase. Co-localization of CDKB2;1-GFP and CycB2;2-GFP on chromosomes aligned at the center of the metaphase cells suggests that the CDKB2-CycB2 complex may function in retaining chromosomes at the metaphase plate. Overexpression of CycB2;2 in rice plants resulted in acceleration of root growth without any increase in cell size, indicating that CycB2;2 promoted cell division probably through association with CDKB2 in the root meristem. PMID- 12753583 TI - Ku80- and DNA ligase IV-deficient plants are sensitive to ionizing radiation and defective in T-DNA integration. AB - Double-strand break (DSB) repair pathways catalyze the rejoining of broken chromosomes and the integration of transforming DNAs. These processes have been well characterized in bacteria, fungi, and animals. Plants are generally thought primarily to utilize a non-homologous end joining (NHEJ) pathway to repair DSBs and integrate transgenes, as transforming DNAs with large tracts of homology to the chromosome are integrated at random. In order to test the hypothesis that NHEJ is an important pathway for the repair of DSBs in plants, we isolated T-DNA insertion mutations in the Arabidopsis homologs of the Ku80 and DNA ligase IV genes, required for the initiation and completion, respectively, of NHEJ. Both mutants were hypersensitive to the cytostatic effects of gamma radiation, suggesting that NHEJ is indeed a critical pathway for the repair of DSBs. T-DNA insertion rates were also decreased in the mutants, indicating that Ku80 and DNA ligase IV play an important role in either the mechanism or the regulation of T DNA integration in Arabidopsis. PMID- 12753584 TI - Identification and expression of a soybean nodule-enhanced PEP-carboxylase kinase gene (NE-PpcK) that shows striking up-/down-regulation in vivo. AB - Various isoforms of plant phosphoenolpyruvate carboxylase (PEPC (Ppc)) are controlled post-translationally by an intricate interaction between allosteric regulation and reversible protein phosphorylation. In leaves and root nodules of legumes, these changes in PEPC phosphorylation state are governed primarily by PEPC-kinase (PpcK), a novel, 'minimal but functional' Ser/Thr kinase. To date, this plant-specific kinase has been investigated in molecular terms exclusively in non-leguminous plants, such as Crassulacean-acid-metabolism (CAM) species and Arabidopsis. As an important extension of our earlier biochemical studies on this dedicated kinase and PEPC phosphorylation in soybean (Glycine max) nodules, we now report the molecular cloning of the first legume PpcK from a soybean nodule cDNA library, which encodes a functional, 31.0 kDa PpcK polypeptide. Besides displaying organ, developmental, and spatial expression properties that are strikingly up-regulated in mature nodules, the expression pattern of this transcript is distinct from that of a second soybean PpcK isogene (GmPpcK). The steady-state abundance of this former, nodule-enhanced transcript (NE-PpcK) is markedly influenced by photosynthate supply from the shoots. This latter up-/down regulation of NE-PpcK transcript level occurs in vivo in concert with the corresponding changes in the nodule PpcK activity, the phosphorylation-state of PEPC, and the abundance of a previously identified, nodule-enhanced transcript (GmPEPC7) that encodes the target enzyme (NE-Ppc). Furthermore, genomic Southern analysis and inspection of the public database indicate that there are at least three distinct PpcK and Ppc isogenes in soybean. Collectively, these and recent findings with Arabidopsis implicate the existence of multiple PpcK-Ppc'expression partners' in plants, exemplified by NE-PpcK and NE-Ppc in the soybean nodule. PMID- 12753585 TI - The FHY3 and FAR1 genes encode transposase-related proteins involved in regulation of gene expression by the phytochrome A-signaling pathway. AB - The Arabidopsis mutants far1 and fhy3 display a phenotype of reduced inhibition of hypocotyl elongation, which is specific to far-red light and therefore specific to the phytochrome A (phyA)-signaling pathway. We report that the proteins encoded by the FAR1 and FHY3 genes are both related to the transposases of type II MuDR family transposons. We demonstrate that the FAR1 protein is capable of activating transcription in Arabidopsis, indicating that it may define a type of transcriptional regulator. Using microarray expression analysis, we show that of 293 mRNAs twofold induced in wild-type Col-0 plants by continuous far-red light, 85% show reduced responsiveness in the fhy3 mutant. Notable alterations were observed in the responses of genes encoding certain transcription factors, proteins involved in cell wall extension, and proteins related to redox balance control. We also found genes, including some involved in transcriptional control, which showed altered transcriptional behavior in the dark-grown mutant plants. Taken together, our data suggest that FAR1 and FHY3 may function 'permissively' outside the signal transduction pathway of light regulated development, yet be required for the expression of transcriptional regulatory components. An alternative possibility is that their role includes both light-signal transduction and transcriptional regulation of other genes not responsive to light. We propose that FAR1 and FHY3 control the expression of their target genes by a mechanism that has evolved directly from the way that an ancestral, MuDRA-like transposase bound to the TIRs of mobile elements. PMID- 12753586 TI - The C-terminal tail of Arabidopsis 14-3-3omega functions as an autoinhibitor and may contain a tenth alpha-helix. AB - The eukaryotic regulatory protein 14-3-3 is involved in many important plant cellular processes including regulation of nitrate assimilation through inhibition of phosphorylated nitrate reductase (pNR) in darkened leaves. Divalent metal cations (Me2+) and some polyamines interact with the loop 8 region of the 14-3-3 proteins and allow them to bind and inhibit pNR in vitro. The role of the highly variant C-terminal regions of the 14-3-3 isoforms in regulation by polycations is not clear. In this study, we carried out structural analyses on the C-terminal tail of the Arabidopsis 14-3-3omega isoform and evaluated its contributions to the inhibition of pNR. Nested C-terminal truncations of the recombinant 14-3-3omega protein revealed that the removal of the C-terminal tail renders the protein partially Mg2+-independent in both pNR binding and inhibition of activity, suggesting that the C-terminus functions as an autoinhibitor. The C terminus of 14-3-3omega appears to undergo a conformational change in the presence of polycations as demonstrated by its increased trypsin cleavage at Lys 247. C-terminal truncation of 14-3-3omega at Thr-255 increased its interaction with antibodies to the C-terminus of 14-3-3omega in non-denaturing conditions, but not in denaturing conditions, suggesting that the C-terminal tail contains ordered structures that might be disrupted by the truncation. Circular dichroism (CD) analysis of a C-terminal peptide, from Trp-234 to Lys-249, revealed that the C-terminal tail might contain a tenth alpha-helix, in agreement with the in silico predictions. The function of the putative tenth alpha-helix is not clear because substituting two prolyl residues within the predicted helix (E245P/I246P mutant), which prevented the corresponding peptide from adopting a helical conformation, did not affect the inhibition of pNR activity in the presence or absence of Mg2+. We propose that in the absence of polycations, access of target proteins to their binding groove in the 14-3-3 protein is restricted by the C terminus, which acts as part of a gate that opens with the binding of polycations to loop 8. PMID- 12753587 TI - Isolation of a glucosyltransferase from Arabidopsis thaliana active in the metabolism of the persistent pollutant 3,4-dichloroaniline. AB - The pollutant 3,4-dichloroaniline (DCA) was rapidly detoxified by glucosylation in Arabidopsis thaliana root cultures, with the N-beta-d-glucopyranosyl-DCA exported into the medium. The N-glucosyltransferase (N-GT) responsible for this activity was purified from Arabidopsis suspension cultures and the resulting 50 kDa polypeptide analysed by matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS) following tryptic digestion. The protein was identified as GT72B1. The GT was cloned and the purified recombinant enzyme shown to be highly active in conjugating DCA and 2,4,5-trichlorophenol, as well as several other chlorinated phenols and anilines, demonstrating both N-GT and O GT activity. GT72B1 showed little activity towards natural products with the exception of the tyrosine catabolite 4-hydroxyphenylpyruvic acid. Both O-GT and N GT activities were enhanced in both plants and cultures treated with herbicide safeners, demonstrating the chemical inducibility of this detoxification system in Arabidopsis. PMID- 12753588 TI - The NFP locus of Medicago truncatula controls an early step of Nod factor signal transduction upstream of a rapid calcium flux and root hair deformation. AB - Establishment of the Rhizobium-legume symbiosis depends on a molecular dialogue, in which rhizobial nodulation (Nod) factors act as symbiotic signals, playing a key role in the control of specificity of infection and nodule formation. Using nodulation-defective (Nod-) mutants of Medicago truncatula to study the mechanisms controlling Nod factor perception and signalling, we have previously identified five genes that control components of a Nod factor-activated signal transduction pathway. Characterisation of a new M. truncatula Nod- mutant led to the identification of the Nod Factor Perception (NFP) locus. The nfp mutant has a novel phenotype among Nod- mutants of M. truncatula, as it does not respond to Nod factors by any of the responses tested. The nfp mutant thus shows no rapid calcium flux, the earliest detectable Nod factor response of wild-type plants, and no root hair deformation. The nfp mutant is also deficient in Nod factor induced calcium spiking and early nodulin gene expression. While certain genes controlling Nod factor signal transduction also control the establishment of an arbuscular mycorrhizal symbiosis, the nfp mutant shows a wild-type mycorrhizal phenotype. These data indicate that the NFP locus controls an early step of Nod factor signal transduction, upstream of previously identified genes and specific to nodulation. PMID- 12753589 TI - Neither inverted repeat T-DNA configurations nor arrangements of tandemly repeated transgenes are sufficient to trigger transgene silencing. AB - Transgene expression was analysed in Arabidopsis T-DNA transformants carrying defined numbers and arrangement of different reporter genes. All transgenes were placed under the control of the strong constitutive CaMV 35S promoter. High, stable transgene expression was observed in plants containing two copies of the beta-glucuronidase (GUS) gene, two or four copies of the green fluorescent protein (GFP) gene and two, four or six copies of the streptomycin phosphotransferase (SPT) gene. Thus, the mere presence of multiple promoter and/or transgene sequences did not result in gene silencing. In none of the cases analysed were tandem repeat arrangements of transgenes and/or inverted repeat (IR) T-DNA structures sufficient to trigger silencing of the different reporter genes. Instead, post-transcriptional gene silencing (PTGS) correlated with the copy number of the highly expressed transgenes. Twelve copies of the SPT and four copies of the GUS gene triggered silencing. Silencing is frequently associated with repetitive T-DNA structures. We favour the idea that in many cases this may be attributed to the high transgene doses rather than the repeat arrangements themselves. PMID- 12753591 TI - Comparative mapping between potato (Solanum tuberosum) and Arabidopsis thaliana reveals structurally conserved domains and ancient duplications in the potato genome. AB - A genetic map of potato (Solanum tuberosum) was constructed based on 293 restriction fragment length polymorphism (RFLP) markers including 31 EST markers of Arabidopsis. The in silico comparison of all marker sequences with the Arabidopsis genomic sequence resulted in 189 markers that detected in Arabidopsis 787 loci with sequence conservation. Based on conserved linkage between groups of at least three different markers on the genetic map of potato and the physical map of Arabidopsis, 90 putative syntenic blocks were identified covering 41% of the potato genetic map and 50% of the Arabidopsis physical map. The existence and distribution of syntenic blocks suggested a higher degree of structural conservation in some parts of the potato genome when compared to others. Syntenic blocks were redundant: most potato syntenic blocks were related to several Arabidopsis genome segments and vice versa. Some duplicated potato syntenic blocks correlated well with ancient segmental duplications in Arabidopsis. Syntenic relationships between different genomic segments of potato and the same segment of the Arabidopsis genome indicated that potato genome evolution included ancient intra- and interchromosomal duplications. The partial genome coveridge and the redundancy of syntenic blocks limits the use of synteny for functional comparisons between the crop species potato and the model plant Arabidopsis. PMID- 12753592 TI - Enhancement of virus-induced gene silencing through viral-based production of inverted-repeats. AB - Plant virus-based vectors carrying sequences homologous to endogenous genes trigger silencing through a homology-dependent RNA degradation mechanism. This phenomenon, called virus-induced gene silencing (VIGS), has potential as a powerful reverse-genetics tool in functional genomic programmes through transient, loss-of-function screens. Here, we describe a method to enhance the robustness of the VIGS phenotype by increasing the level of dsRNA molecule production, a critical step in the VIGS response. Incorporation of 40-60 base direct inverted-repeats into a plant viral vector generates RNA molecules that form dsRNA hairpins. A tobacco mosaic virus (TMV)-based vector carrying such inverted-repeats, homologous to a green fluorescent protein (gfp) transgene or an endogenous phytoene desaturase (pds) gene, generated a stronger and more pervasive VIGS phenotype than constructs carrying corresponding cDNA fragments in sense or antisense orientation. Real-time RT-PCR indicated that there was up to a threefold reduction in target mRNA accumulation in the tissues where VIGS was triggered by constructs carrying inverted-repeats compared to those where it was triggered by sense or antisense constructs. Moreover, an enhanced VIGS pds phenotype was observed using a different vector, based on barley stripe mosaic virus, in the monocotyledonous host barley. This demonstrates that VIGS can be significantly improved through the inclusion of small inverted-repeats in plant virus-based vectors, generating a more robust loss-of-function phenotype. This suggests that dsRNA formation can be a limiting factor in the VIGS phenomenon. PMID- 12753590 TI - Disruption of Arabidopsis thaliana MYB26 results in male sterility due to non dehiscent anthers. AB - A male sterile mutant with a defect in anther dehiscence was identified in an Arabidopsis thaliana population mutagenized with the Zea mays transposon En 1/Spm. Mutants produce viable pollen that can fertilize when released mechanically from the anthers. Mutant stamens are of normal size and shape, but lack cell wall fortifications in the endothecial cell layer of the anther, which are required for the dehiscence process. The mutant phenotype was shown to be caused by a transposon insertion in AtMYB26, disrupting the putative DNA-binding domain of this R2R3-type MYB transcription factor. RT-PCR revealed that expression of AtMYB26 is restricted to inflorescences. Sterility was shown to be stable under several environmental conditions. The high stability of the sterile phenotype, together with the fact that pollen is functional, makes AtMYB26 and its orthologs a valuable tool for manipulating male fertility in higher plants. PMID- 12753593 TI - Nanolitre-scale assays to determine the activities of enzymes in individual plant cells. AB - There are a variety of methods for characterising gene expression at the level of individual cells and for demonstrating that the cells also contain the encoded proteins. However, measuring the activity of enzymes at the resolution of single cells in complex tissues, such as leaves, is problematic. We have addressed this by using single-cell sampling to extract 10-100 pl droplets of sap from individual plant cells and then measuring enzyme activities in these droplets with nanolitre-scale fluorescence-based assays. We have optimised these assays and used them to measure and characterise the activities of acid phosphatase, cysteine protease and nitrate reductase in sap samples from epidermal and mesophyll cells of barley (Hordeum vulgare L.) and Arabidopsis thaliana leaves exposed to different developmental and environmental conditions. During leaf senescence in barley, we found that the dynamics with which acid phosphatase and protease activities changed were different in each cell type and did not mirror the changes occurring at the whole-leaf level. Increases in nitrate reductase activities after exposure of barley plants to nitrate were large in mesophyll cells but small in epidermal cells. The technique was applied successfully to Arabidopsis and, as in barley, revealed cell-specific differences in the activities of both acid phosphatase and nitrate reductase. The assays add to the spectrum of techniques available for characterising cells within complex plant tissues, thus extending the opportunity to relate gene expression to biochemical activities at the single-cell level. PMID- 12753594 TI - Where will the wound healing clinicians and researchers of the future be developed? PMID- 12753595 TI - Impediments to drug development. AB - There is a continual need for new products for wound care, as well as a desire by scientists and clinicians to translate information into wound healing improvements for patients. Pharmaceutical and biotechnology companies devote immense resources to fulfilling these needs and desires. However, there are many impediments to drug development that are poorly understood by caregivers, patients, and the public at large. Among these impediments are the tremendous costs involved, the short patent protection time, and regulatory issues. In addition, there is a marked attrition of potential drugs as they progress through the various stages of development. When the costs, time, regulatory issues, and attrition impediments are overcome, the problems with reimbursement become an impediment. This is especially true in the elderly population in which most chronic wound healing problems occur. Finally, academic societies such as the Wound Healing Society and its members pose an impediment to drug development. There is a need to interact with various governmental agencies and industry to facilitate translating science to patient care. This has not been done with a strong, uniform voice. These are but a few of the impediments that prevent scientific advances from resulting in new products available at the bedside to improve the quality of life of our patients. PMID- 12753597 TI - Evidence of oxidative stress in chronic venous ulcers. AB - Reactive oxygen species have been implicated in the impaired healing of chronic leg ulcers but little direct evidence is available. We have observed a significant (p < 0.01) elevation of the allantoin : uric acid percentage ratio, a marker of oxidative stress, in wound fluid from chronic leg ulcers (median 17, range 8-860) compared to both paired plasma (median 2, range 1-8) and acute surgical wound fluid (median 4, range 3-7). However, the allantoin : uric acid percentage ratio did not differ significantly between chronic wounds that healed and those that failed to heal. Neutrophil elastase was elevated 30- to 1300-fold in chronic wound fluid compared to plasma and there was a correlation (r(2) = 0.742) between wound fluid elastase and the allantoin : uric acid percentage ratio. Total antioxidant capacity of wound fluid, as measured with a chemiluminescence assay, did not show a correlation (r(2) = 0.03) with the observed oxidative stress. These observations suggest that conditions of localized oxidative stress, possibly related to neutrophil-associated production of reactive oxygen species, are present in chronic leg ulcers. It is possible that future therapeutic strategies aimed at reducing oxidative stress, in addition to good standard care, could improve healing rates of chronic wounds. PMID- 12753596 TI - Randomized trial of four-layer and two-layer bandage systems in the management of chronic venous ulceration. AB - To compare a four-layer bandage system with a two-layer system in the management of chronic venous leg ulceration, a prospective randomized open parallel groups trial was undertaken. In total, 112 patients newly presenting to leg ulcer services with chronic leg ulceration, screened to exclude the presence of arterial disease (ankle brachial pressure index <0.8) and causes of ulceration other than venous disease, were entered into the trial. Patients were randomized to receive either four-layer (Profore) or two-layer (Surepress) high-compression elastic bandage systems. In all, 109 out of 112 patients had at least one follow up. After 24 weeks, 50 out of 57 (88%) patients randomized to the four-layer bandage system with follow-up had ulcer closure (full epithelialization) compared with 40 out of 52 (77%) on the two-layer bandage, hazard ratio = 1.18 (95% confidence interval 0.69-2.02), p = 0.55. After 12 weeks, 40 out of 57 (70%) patients randomized to the four-layer bandage system with follow-up had ulcer closure compared with 30 out of 52 (58%) on the two-layer bandage, odds ratio = 4.23 (95% confidence interval 1.29-13.86), p = 0.02. Withdrawal rates were significantly greater on the two-layer bandage (30 out of 54; 56%) compared with the four-layer bandage system (8 out of 58; 14%), p < 0.001, and the number of patients with at least one device-related adverse incident was significantly greater on the two-layer bandaging system (15 out of 54; 28%) compared with four layer bandaging (5 out of 54; 9%), p = 0.01. The higher mean cost of treatment in the two-layer bandaging system arm over 24 weeks ($1374 [ pound 916] vs. $1314 [ pound 876]) was explained by the increased mean number of bandage changes (1.5 vs. 1.1 per week) with the two-layer system. In conclusion, the four-layer bandage offers advantages over the two-layer bandage in terms of reduced withdrawal from treatment, fewer adverse incidents, and lower treatment cost. PMID- 12753598 TI - Matrix metalloproteinases and their tissue inhibitors in severely burned children. AB - Severe burns cause not only skin injury but several marked systemic derangements. During wound healing, matrix metalloproteinases (MMPs) and the tissue inhibitors of metalloproteinases play an important role in tissue regeneration and remodeling processes. Therefore, in the present study, we determined the serum levels of MMPs and tissue inhibitor of metalloproteinase-1 in burn patients over time. Serum samples from 12 severely burned children (mean age 7.9 +/- 2.5 years) with >40% total body surface area burns were obtained within 0.5 hours, 3, 7, and 21 days after injury. Pro-MMP-1, MMP-3, MMP-9, and tissue inhibitor of metalloproteinase-1 serum levels were assayed by enzyme-linked immunoassay and compared to normal healthy volunteers. Two-way analysis of variance and Bonferroni's test were used for statistical analysis. Pro-MMP-1 levels in the serum were significantly elevated by the seventh day after burn. MMP-3 and MMP-9 levels showed significant increases by day 3 and 21 compared to normals, respectively. Tissue inhibitor of metalloproteinase-1 levels did not change with time after burn but were significantly higher by 3 days after burn compared to normal serum. In conclusion, changes in MMPs and tissue inhibitor of metalloproteinase-1 occur in burn patients and those changes may be a mechanism beneficial to wound healing. PMID- 12753599 TI - Evaluation of a new liquid occlusive dressing for excisional wounds. AB - We evaluated a novel octylcyanoacrylate-based liquid occlusive dressing for partial-thickness wounds. One hundred and fifteen standardized wounds were created with an electric dermatome set at a depth of 600 micro on the flanks of three pigs and randomly treated with liquid occlusive dressing, a hydrocolloid dressing, or gauze. In one pig, wounds were swabbed with Staphylococcus aureus. Biopsies were taken after 4, 5, 6, and 21 days. Hemostasis was obtained in all wounds treated with the liquid occlusive. The percent reepithelialization of wounds treated with the liquid occlusive and hydrocolloid dressings were significantly greater at days 4 and 5 than control wounds (78% and 82% vs. 40%, p < 0.001 and 99% and 100% vs. 72%, p < 0.001, respectively). None of the liquid occlusive-treated wounds challenged with bacteria became infected. Foreign body reactions were least common in wounds treated with the liquid occlusive (p < 0.001). Scar depth was less for liquid occlusive- and hydrocolloid-treated wounds than controls (285 micro and 303 micro vs. 490 micro, p < 0.001). We conclude that excisional wounds treated with the liquid occlusive dressing reepithelialize as quickly as hydrocolloid-treated wounds. The liquid occlusive dressing is an effective microbial barrier and hemostatic agent resulting in fewer foreign body reactions than hydrocolloid-treated wounds or controls. PMID- 12753600 TI - Topical epiregulin enhances repair of murine excisional wounds. AB - Epiregulin is a broad specificity epidermal growth factor family member that activates ErbB1 and ErbB4 homodimers and all possible heterodimeric ErbB complexes. Our objective was to determine whether topical epiregulin enhanced repair of murine excisional wounds. Wounds were treated on days 0-4 with either topical epiregulin (1 micro g/ml), epidermal growth factor (10 micro g/ml), or vehicle. At day 5 postinjury, wounds receiving epiregulin were significantly smaller than those treated with epidermal growth factor or vehicle. Treatment with epiregulin promoted greater epidermal proliferation and thickening than epidermal growth factor or vehicle due to an expansion of the proliferative compartment of keratinocytes. Dermal thickness was also increased in epiregulin treated wounds as compared to those treated with epidermal growth factor or vehicle. In day 5 wounds, matrix metalloproteinase-3 (stromelysin-1) mRNA levels were significantly lower in epiregulin- or epidermal growth factor-treated wounds than in vehicle-treated controls, suggesting that growth factor-treated wounds were more mature and required less ongoing proteolytic activity than their same day vehicle-treated counterparts. This is the first report that topical epiregulin accelerates repair of full-thickness murine excisional wounds as compared to vehicle or epidermal growth factor. Furthermore, epiregulin is more potent and more effective than epidermal growth factor in promoting proliferation and maturation of the epidermis as well as enhancement of the neodermis. PMID- 12753601 TI - Supplemental L-arginine enhances wound healing in diabetic rats. AB - L-arginine has been shown to enhance wound strength and collagen deposition in rodents and humans. Diabetes mellitus, which impairs wound healing, is accompanied by a reduction in nitric oxide at the wound site. The amino acid L arginine is the only substrate for nitric oxide synthesis. We sought to determine whether supplemental L-arginine can restore the impaired wound healing of diabetic rats. Fifty-six male Lewis rats were used in this study, of which twenty nine rats were rendered diabetic 7 days prior to surgery with intraperitoneal streptozotocin. Twenty-seven untreated rats served as controls. Animals underwent a dorsal skin incision with implantation of polyvinyl-alcohol sponges. Sixteen diabetic and 14 normal rats received 1 g/kg/day of L-arginine by injection, while the remainder received saline injections only. Animals were euthanized 10 days postwounding, and their wounds were analyzed for breaking strength. The wound sponges were assayed for total hydroxyproline and nitrite/nitrate content. Plasma and wound fluid concentrations of L-arginine, ornithine, and citrulline were determined. Wound sponge RNA was extracted and subjected to Northern blot analysis for procollagen I and III. Diabetic wounds had greatly decreased breaking strengths compared with controls. L-arginine significantly enhanced wound breaking strengths in both control (+23%) and diabetic animals (+44%), and also increased wound hydroxyproline levels in both diabetic (+40%) and control animals (+24%) as compared to their saline-treated counterparts. mRNA for procollagen I and III were elevated by L-arginine treatment in both diabetic rats and controls. Treatment with L-arginine significantly increased wound fluid nitrite/nitrate levels in diabetic animals. The data show that the impaired healing of diabetic wounds can be partially corrected by L-arginine supplementation, and that this effect is accompanied by enhanced wound nitric oxide synthesis. PMID- 12753602 TI - Topical vanadate optimizes collagen organization within granulation tissue. AB - Systemic ingestion of vanadate, a nonspecific inhibitor of tyrosine phosphatases, doubles wound breaking strength, enhances the packing of collagen fibers, and prevents the appearance of myofibroblasts in granulation tissue. Will the local application of vanadate mimic the systemic effects? Pairs of polyvinyl alcohol sponges, each with a central reservoir and attached injection port, were subcutaneously implanted in rats. Daily, one implant received 0.2 ml of saline and the other received 0.2 ml of 0.03 mM vanadate in saline. On day 7, harvested sponges had equivalent wet weights. The vanadate-treated sponges had fibroblasts separated by connective tissue, with a more intense birefringence of the collagen fibers. Transmission electron microscopy showed collagen more uniformly packed in the vanadate treated sponges where collagen fibers were equally spaced and had equal diameters. By immunohistology, myofibroblasts, defined by the expression of alpha-smooth muscle actin within stress fibers, were absent in vanadate-treated granulation tissue. The expression of alpha-smooth muscle actin was restricted to smooth muscle cells of blood vessels. Controls had densely packed alpha-smooth muscle actin staining myofibroblasts, weak birefringence, and randomly spaced collagen fibers with irregular diameters. We conclude that the local application of vanadate prevents the appearance of myofibroblasts and optimizes the organization of collagen fibers in developing granulation tissue. PMID- 12753603 TI - Impaired healing of nitrogen mustard wounds in CXCR2 null mice. AB - To examine the significance of chemokine activation of CXCR2 in wound healing after chemical burn, cutaneous injury was created by topical application of nitrogen mustard on CXCR2 wild type (+/+), heterozygous (+/-), and knockout (-/-) mice. Wounds were analyzed histologically for neutrophil and monocyte infiltration and for reepithelialization at postwound days 4, 7, and 10. Neutrophil recruitment to the wound site was reduced through postwound day 7 in CXCR2 -/- mice as indicated by myeloperoxidase assay and by visual quantitation. Because there is always concern that mice with targeted deletion of a specific receptor may undergo developmental adaptations to offset the loss of the receptor, we also accessed chemical wound repair in the presence of a small molecule antagonist of CXCR2. Dietary supplementation with a CXCR2 antagonist (SB 265610) during the wound repair process also markedly delayed healing parameters in CXCR2 +/+ mice, even greater than treatment with glucocorticoids. These parallel studies further establish that mice deficient in CXCR2 function exhibit delayed cutaneous wound healing that may be primarily linked to impaired neutrophil recruitment after chemical burn with nitrogen mustard. Thus, there may be a potential therapeutic benefit of treating nitrogen mustard-induced skin lesions with agonists of CXCR2 to facilitate the wound repair process. PMID- 12753604 TI - Recombinant connective tissue growth factor modulates porcine skin fibroblast gene expression. AB - Connective tissue growth factor (CTGF) is a 38 Kda cysteine-rich, heparin-binding peptide that has been implicated in several normal and abnormal physiological processes. CTGF has been shown to be induced by transforming growth factor-beta. Previous studies in our pig model of skin wound healing showed a coordinate expression of transforming growth factor-beta and CTGF during the healing process. To better understand the function of CTGF during wound healing, normal porcine fibroblasts were isolated from skin samples from SPF Yorkshire pigs. At fourth passage the cells were cultured in Dulbecco's modified Eagle's medium supplemented with fetal calf serum and at 80% confluence the medium was replaced with supplemented serum-free medium. After a further 24 hours, cells were treated with 0, 10, 25, 50, 100, and 500 ng/ml of 38 Kda or 16-20 Kda (C-terminal truncated form) recombinant expressed human CTGF for 24 hours or treated with 100 ng/ml for 0, 12, 24, and 48 hours. Subsequently, CTGF effects on cell DNA synthesis and mRNA levels for a subset of relevant molecules were assessed. The results showed that in cells treated with 38 Kda rhCTGF, mRNA levels for types I and III collagen, fibromodulin, and basic fibroblast growth factor were significantly up-regulated, but mRNA levels for HSP47, decorin, biglycan, and versican were not significantly altered. mRNA levels for CTGF were also significantly increased, indicating autoregulation of expression. However, mRNA levels for transforming growth factor-beta, inteleukins 1 and 6, tumor necrosis factor-alpha, and nerve growth factor did not change. Interestingly, mRNA levels for the tissue inhibitors of metalloproteinase-1, -2, -3 and -4 were observed to significantly increase, but in contrast, mRNA levels for matrix metalloproteinases-1, -2, -9 were not significantly altered by exposure of the cells to the 38 Kda form of CTGF. In addition, DNA synthesis was augmented in the presence of 38 Kda rhCTGF. However, the truncated 16-20 Kda form of rhCTGF appeared to have none of these effects on porcine fibroblasts. These results indicate that in order to induce changes in porcine fibroblasts a molecule with an intact C-terminal domain is required, and that CTGF regulates porcine fibroblast extracellular matrix molecule, growth factor, and proteinase inhibitor gene expression without apparently affecting matrix metalloproteinase mRNA levels. These findings suggest that CTGF contributes to the anabolic environment during skin wound healing via selective modulation of fibroblast proliferation and changes to gene expression. PMID- 12753605 TI - Examination gloves affect secretion of matrix metalloproteinases and their inhibitors from human abdominal skin fibroblasts. AB - Overexpression of matrix metalloproteinases (MMPs) has been observed in chronic, compared to acute, wounds and altered levels might impair healing. During treatment of wounds, examination gloves are routinely used, and the wound environment thus gets exposed to gloves. The aim of this study was to characterize secretion of MMPs and tissue inhibitors of metalloproteinases (TIMPs) in cultured fibroblasts with or without exposure to gloves. Cultures were exposed to glove washings from powdered or powder-free latex examination gloves and compared to untreated controls. MMP-1, -2, -3, -9 and their inhibitors TIMP-1 and -2 were assayed in conditioned media. Cells exposed to gloves reduced their release of MMP-1, -2, and -3 with no differences between the manufacturers of the gloves. The inhibitor TIMP-1 was reduced to 10-15% of untreated control values (p < 0.001), being less affected by the powder-free than by the powdered glove (p < 0.05). MMP-9 and TIMP-2 were not significantly altered. We therefore conclude that secretion of MMPs and TIMPs from cultured fibroblasts were affected by glove washings. Powdered and powder-free gloves had similar effects, except for a less pronounced reduction of TIMP-1 production by the powder-free glove. Examination gloves might therefore affect wound healing, with the least pronounced effect observed using the powder-free glove. PMID- 12753606 TI - Silicone sheet for treatment and prevention of hypertrophic scar: a new proposal for the mechanism of efficacy. PMID- 12753608 TI - Staphyloma in a cat. AB - A unilateral scleral staphyloma in an 18-month-old, female spayed Domestic Short haired cat was treated with excision, primary closure and fascial graft. Other ocular abnormalities noted on examination included iris coloboma, anterior cortical cataract, focal lens equator flattening and retinal dysplasia. The staphyloma was presumed to be congenital in origin. PMID- 12753609 TI - Keratitis due to Histoplasma spp. in a horse. AB - A 5-year-old Holsteiner gelding from Germany was presented 2 months after a whitish discoloration of the left cornea was observed. Cytologic examination revealed intra- and extracellular globular structures, up to 4 micro m in size, consisting of a central spherical deeply basophilic body surrounded by an unstained halo. The structures were morphologically consistent with Histoplasma spp. Infection with Histoplasma organisms is not endemic in Europe. Topical use of fluconazole was successful in eliminating Histoplasma organisms within 10 days of initiation of treatment. PMID- 12753610 TI - Intraocular pressure development after cataract surgery: a prospective study in 50 dogs (1998-2000). AB - OBJECTIVE: To study the course of intraocular pressure (IOP) after cataract surgery in 50 dogs. DESIGN: Prospective study. ANIMALS: Fifty dogs without preoperative ocular hypertension were selected for cataract surgery. METHODS: All dogs underwent cataract surgery: 25 by manual extracapsular extraction and 25 by phacoemulsification. For each dog, intraocular pressure was measured before surgery, and 1, 3, 5, 18 h, 1 week and 1 month post surgery. RESULTS: No significant difference of mean intraocular pressure between the two surgical methods was observed for each time measurement. Nine dogs had postoperative hypertension (IOP > 25 mmHg) during the first 5 hours post surgery. Incidence of postoperative hypertension was not significantly different with manual extracapsular extraction (16%) vs. phacoemulsification (20%). A decrease of mean IOP was observed 1 h after surgery (8.49 mmHg vs. 10.91 mmHg), then an increase 3 and 5 h post surgery (12.3 and 13.32 mmHg, respectively). At 18 h, 1 week and 1 month post surgery, mean IOP decreased. Mean IOP was 10.38, 10.38 and 8.84 mmHg, respectively. CONCLUSION: In this study incidence of POH is not high. However, a follow-up of IOP in the first hours after cataract surgery is required to avoid complications of the retina and optic nerve and to administer hypotensive treatment if necessary. PMID- 12753611 TI - The use of transscleral cyclophotocoagulation with a diode laser for the treatment of glaucoma occurring post intracapsular extraction of displaced lenses: a retrospective study of 15 dogs (1995-2000). AB - The objective of this retrospective study was to evaluate transscleral cyclophotocoagulation (TSCP) using a diode laser for treating aphakic glaucoma that developed after intracapsular lens extraction (ICLE) had been performed for displaced lenses. Records of 15 dogs (21 eyes) were reviewed. The intraocular pressure (IOP) and the presence of vision were recorded at the time of ICLE and TSCP and at 1, 3, 6-9, 12 and 24 months post TSCP. The glaucoma was considered controlled if the IOP was less than 25 mmHg. The results indicated that the effectiveness of TSCP using a diode laser for treating aphakic glaucoma was of a short-term duration (1-3 months), with three patients needing repeat TSPC. In addition, adjunctive antiglaucoma medications were often required to maintain an adequately controlled IOP. Over the 24-month period the number of dogs requiring intrascleral prosthesis, intraocular gentamicin or enucleation, or that were lost to follow-up, increased. PMID- 12753612 TI - Use of a caudal auricular axial pattern flap in three cats and one dog following orbital exenteration. AB - Orbital exenteration accompanied by wide eyelid excision in the cat and dog may leave a defect that cannot be closed in a primary fashion. This report describes the use of a caudal auricular axial pattern flap to effect closure following orbital exenteration in three cats and one dog. The most common complication was distal flap necrosis, which necessitated a second surgery in two patients. PMID- 12753613 TI - Day-blindness in three dogs: clinical and electroretinographic findings. AB - A 6-month-old Rhodesian ridgeback-cross, a 6-year-old Chihuahua and a 12-month old Australian cattle dog were presented to the authors with a history of colliding with obstacles in daylight. Ophthalmic examination was normal and all three dogs successfully negotiated obstacle courses in dim light. In daylight the dogs became suddenly blind and repeatedly collided with obstacles. Elecroretinography (ERG) revealed no retinal activity to high frequency (30 Hz), bright intensity blue light retinal stimulation by any dog, confirming cone dysfunction. Achromatopsia has previously been recorded in Alaskan malamutes and miniature poodles. This clinical case series illustrates the characteristic behavioral presentation and the electroretinographic findings of severe day blindness and demonstrates that this condition may exist in other breeds of dogs. PMID- 12753614 TI - Presence of opioid growth factor and its receptor in the normal dog, cat and horse cornea. AB - OBJECTIVE: To determine if opioid growth factor (OGF, [Met5]enkephalin) and its specific receptor (OGFr) are present in normal cat, dog and horse cornea. ANIMALS STUDIED: Normal dog, cat and horse. PROCEDURE: Corneas were obtained from animals euthanized for reasons unrelated to this project. One cornea from each of three normal cats, dogs and horses was evaluated. The right or left cornea from each animal was chosen randomly. Corneas were harvested and placed in corneal storage media for transport to The M.S. Hershey Medical Center of The Pennsylvania State University where immunocytochemistry techniques were used to demonstrate the presence and location of OGF and OGFr. Tissues were rinsed in Sorenson's phosphate buffer, immersed in 20% sucrose in buffer and then snap frozen in isopentane. Corneas were then embedded in OCT medium and 15 micro m cryostat sections were created. Presence of OGF was determined by using a polyclonal antibody to [Met5]enkephalin and assessing immunoreactivity. OGFr presence was determined by using a previously characterized rabbit polyclonal antibody to the receptor. RESULTS: OGF and OGFr were identified in large quantities in the corneal epithelium of all three species. CONCLUSION: Opioid growth factor and its specific receptor are present in the corneal epithelium of normal cats, dogs and horses. OGF is present in the cornea of many species and its presence is theorized to inhibit healing of injured tissue. PMID- 12753615 TI - Lens sparing pars plana vitrectomy and retinal transplantation in cats. AB - Neuroretinal transplantation techniques have been evolving during recent years. Experiments in rodent models with degenerative retinal disease have been encouraging. This paper describes a surgical technique developed for use in the Abyssinian cat mutant. After two-port pars plana vitrectomy, retinotomy and bleb formation, whole sheets of neonatal neuroretinal allografts were placed into the subretinal space. The surgery was difficult but feasible, and the main complication was intraoperative hemorrhage. PMID- 12753616 TI - The effect of an L-type calcium channel blocker on the hemodynamics of orbital arteries in dogs. AB - PURPOSE: (1) To determine the effect of the l-type calcium channel blocker amlodipine on color Doppler ultrasound-determined vascular resistance and blood flow velocities in the distal retrobulbar arteries of dogs; (2) to determine any effect of blood pressure and PCO2 rate on such color Doppler-determined circulatory measurements. METHODS: Color Doppler imaging measurements of the short posterior ciliary artery, long posterior ciliary artery, and ophthalmic artery of normal eyes of 10 dogs were obtained under isofluorane anesthesia before and 1 week after oral amlodipine administration. Mean systemic arterial blood pressure and PCO2 were monitored. RESULTS: The mean resistive index decreased significantly in the short posterior ciliary artery (P = 0.0347), in the long posterior ciliary artery (P = 0.0092), and ophthalmic artery (P = 0.0004) following systemic amlodipine administration. The end diastolic velocity increased significantly in the long posterior ciliary artery (P = 0.0368) and ophthalmic artery (P < 0.0001). The peak systolic velocity increased significantly in the ophthalmic artery (P = 0.0256). Mean systemic arterial blood pressure was significantly negatively associated with resistive index (P < 0.0001) and significantly associated with the log of the end diastolic velocity (P < 0.0001). CONCLUSIONS: Systemically administered amlodipine increases color Doppler imaging-determined blood flow velocity and decreases vascular resistive index in the ophthalmic artery, short posterior ciliary artery and long posterior ciliary artery of normal dogs. Changes in systemic arterial blood pressure can significantly affect the measurement of color Doppler imaging parameters. PMID- 12753618 TI - Corneal esthesiometry in the healthy horse. AB - OBJECTIVE: To determine corneal sensitivity in healthy adult horses in order to establish reference values. ANIMALS STUDIED: One hundred eyes of 50 healthy adult horses. PROCEDURE: Corneal sensitivity was determined by evaluating the corneal touch threshold (CTT) in five different corneal regions using a Cochet-Bonnet esthesiometer. RESULTS: Comparing CTT values (in mm filament length) of the five prescribed corneal regions revealed regional variations in corneal sensitivity, with the central region (21.15 +/- 6.23 mm) being the most sensitive, followed by the nasal (20.75 +/- 5.14 mm), temporal (20.70 +/- 5.37 mm), ventral (20.15 +/- 5.88 mm) and dorsal (15.85 +/- 3.70 mm) region. However, differences between the central, nasal, temporal and ventral regions were not statistically significant, whereas the dorsal region was significantly (P < 0.0001) less sensitive compared to the other regions. Division of the horses into three age groups revealed an insignificant decrease in corneal sensitivity with age in the nasal, temporal, ventral and central region, and an insignificant increase with age in the dorsal region. Correlation of corneal sensitivity with age was statistically significant in the central, ventral and temporal region. No statistically significant difference in the CTT was found between left and right eyes and between males and females. CONCLUSIONS: CTT values in 100 eyes of 50 healthy adult horses evaluated with a Cochet-Bonnet esthesiometer generated reference values. Reference values are necessary to determine alterations of corneal sensation accompanying various eye diseases or systemic diseases. PMID- 12753617 TI - Effects of twice daily application of 2% dorzolamide on intraocular pressure in normal cats. AB - The effect of the topical carbonic anhydrase inhibitor, dorzolamide, on intraocular pressure in cats was studied. The intraocular pressure of both eyes of eight healthy cats was measured using applanation tonometery (Tono-Pen) during two phases: control and treatment. Both eyes were treated with 2% dorzolamide twice a day during the treatment phase. Application of dorzolamide resulted in a significant decrease of intraocular pressure over the treatment phase. By the fourth and fifth day of treatment, the mean intraocular pressure (+/- standard deviation) was 9.7 (+/- 1.5) mmHg, whereas the mean (+/- standard deviation) for the last 2 days of the control period was 12.2 (+/- 2.0) mmHg. Based on this study, dorzolamide is a potentially effective treatment for glaucoma in cats, significantly lowering intraocular pressure. PMID- 12753619 TI - IgA and secretory component (SC) in the third eyelid of domestic animals: a comparative study. AB - OBJECTIVE: The third eyelid of domestic animals is important for the production and distribution of tears, in removing ocular debris and in protection of the globe, and has significant immunologic functions. Although it is known that tears contain antibodies of the immunoglobulin A (IgA) isotype which are produced mainly by plasma cells of the lacrimal gland, very little is known about the antibody repertoires in the third eyelid of domestic animals. To assess whether IgA is derived from local synthesis, we analyzed the location of IgA-producing cells and the cellular distribution of secretory component (SC) in the third eyelid of domestic animals in a comparative study. ANIMAL STUDIED: A total of 83 third eyelids of dogs, cats, pigs, cows, sheep, goats and horses were investigated in the course of this study. PROCEDURES: Third eyelids were obtained immediately after death, cut length-wise, fixed overnight and processed for immunohistochemical detection of IgA and SC by the ABC technique. RESULTS: The results show that IgA-producing plasma cells are densely populated in subepithelial spaces of the surface epithelium as well as in the nictitating gland in a species-specific manner. In contrast, the SC could be demonstrated exclusively in glandular acinar and ductal epithelial cells and in different cell types of the surface epithelium, preferentially located on the bulbar side of the nictitating membrane. CONCLUSION: It is suggested that most of the SC is locally produced by resident plasma cells and subsequently transferred through the surface epithelium and glandular duct cells by transcytosis. This indicates that the third eyelid is an important member of the secretory immune system in domestic animals. PMID- 12753620 TI - Idiopathic ocular and nasal granulomatous inflammatory disease in a dog. AB - A young female Akita presented for ocular and nasal lesions. Examination showed bilateral 360 degree limbal masses that extended onto the peripheral cornea. In addition, intranasal granulomatous masses were observed by rhinoscopy. Histopathologic examination with immunohistochemical staining showed that these masses were almost identical and were classified as an idiopathic, T-cell rich, granulomatous inflammation. The dog responded well to anti-inflammatory medication and the disease remains in remission 2 years after the initial presentation. PMID- 12753621 TI - Unilateral ocular subalbinism in a laboratory Beagle dog. AB - Blue discoloration of the iris was found in the left eye of a male laboratory Beagle dog, which had a normal tricolor coat and clinically showed no visual impairment. Ophthalmoscopically, the affected eye revealed red-colored tigroid fundus, in which no tapetum was present. The retinal vasculature and the optic disc showed no noticeable changes. Histopathologically, in the left eye melanocytes had extremely few melanin granules in the anterior segment, including the anterior border layer, in the stroma and sphincter muscle of the iris and in the stroma of the ciliary body and choroid. However, the posterior pigment epithelium of the iris, the pigment epithelium of the ciliary body and the retinal pigment epithelium showed normal pigmentation. The tapetal elements were completely absent. Number and distribution of the S-100 protein-positive melanocytes with or without melanin granules in the iris, ciliary body and choroid of the left eye were similar to those of the normal right eye. Ultrastructurally, melanocytes in the anterior segment of the affected iris possessed no or few melanosomes which were incompletely melanized. In the right eye, no abnormal features were observed. Based on these results, the present case was diagnosed as unilateral ocular subalbinism with tapetal aplasia in a Beagle dog. PMID- 12753622 TI - Myxoid leiomyoma of the iris in a dog. AB - A leiomyoma of the iris is described in an 11-year-old Yorkshire Terrier. This is a rare primary intraocular tumor in dogs and we describe the clinical presentation, gross findings and histopathologic characteristics of this tumor. The diagnosis was made on the basis of light microscopy and immunohistochemical staining using antidesmin antibodies, which is specific for myogenic tissues. An unusual feature of the tumor was the presence of myxoid change. To our knowledge myxoid change has not been previously described in a primary intraocular leiomyoma. PMID- 12753623 TI - Intraocular extramedullary plasmacytoma in a cat. AB - An 8-year-old, castrated male Domestic Short-haired cat was referred for evaluation of a possible intraocular neoplasm following previous ocular trauma. The eye was blind, and uveitis and an iridal mass were noted on examination. An enucleation was performed and the mandibular lymph node excised. Histopathologic examination revealed neoplastic proliferation of plasma cells in the iris and lymph node. No other evidence of disseminated disease was detected. This is the first case reported of an intraocular extramedullary plasmacytoma in the cat. The variation in clinical manifestations and potential association with multiple myeloma are not known at this time. Disseminated metastasis from a primary plasmacytoma of the uveal tract could also involve the bone marrow and be indistinguishable from multiple myeloma. Early enucleation, as in trauma associated sarcomas, may be indicated to prevent metastasis. Periodic systemic evaluation for evidence of multiple myeloma should be performed. PMID- 12753624 TI - Abstracts of the International Society for Pharmacoeconomics and Outcomes Research 8th Annual International Meeting. May 18-21, 2003. Arlington, Virginia, USA. PMID- 12753629 TI - Editorial: giving antiretrovirals in the peripartum period to prevent mother-to child HIV transmission in low-income countries: only a short-term stopgap measure. PMID- 12753630 TI - An evaluation of antiretroviral HIV/AIDS treatment in a Rio de Janeiro public clinic. AB - The Brazilian public health system has implemented free, universal access to antiretroviral (ARV) therapy for HIV-infected patients. To evaluate this system, we performed a pilot study to determine whether ARVs were prescribed according to Brazilian guidelines in place in 2000, and whether prescriptions were refilled in a timely manner. Year 2000 data were abstracted from all medical and pharmacy records of adult patients first registered for HIV/AIDS care in a Rio de Janeiro public clinic from January to June 2000 (n = 67). Results were analysed using frequency analyses, chi-square tests and logistic regression. The patient sample was 41.8% female and had a mean age of 34.9 years. 54 (81%) had AIDS; total sample mean baseline CD4+/viral counts were 276 cells/mm3 and 237 517 copies per millilitre, respectively. Delays between clinic request and receipt of first CD4+/viral load results ranged from 25 to 107 (mean 66) and 33 to 139 (mean 86) days, respectively. Fifty-nine patients (88.1%) were prescribed ARV treatment. Forty-two regimens (71.2%) were highly active antiretroviral therapies; 17 (28.8%) were combination regimens with two nucleoside reverse transcriptase inhibitors. No combinations were prescribed that were contraindicated in Brazilian guidelines, however 33 patients (55.9%) were prescribed ARV drugs before one or both HIV status parameters (initial CD4+ level or viral load) were recorded. Fourteen patients prescribed ARVs (23.7%) lacked a supply of medication for >1 month during the year at least once. Of these patients, 11 had treatment lapses as a result of failure to pick up medications, and three lacked medication because of drug shortages. Medication lapses were associated with female sex, being hospitalized in 2000, and having more than two drugs in regimen, but were not associated with age, CD4+ level or use of ARVs before 2000. The results from this pilot study suggest conservative prescription of HAART, high practitioner adherence to guidelines, and some problems with refilling medications in a timely manner. Monitoring delays were identified as a structural limitation to optimal adherence to practice guidelines. Better access to monitoring-laboratory facilities and greater drug availability would improve programme success. PMID- 12753631 TI - Polio eradication--the validity of surveillance indicators. AB - A major pre-requisite for polio-free certification by the World Health Organization is that the local surveillance system successfully detects one case of non-polio acute flaccid paralysis (AFP) per 100,000 children below 15 years of age per annum and that no cases of polio occur for three consecutive years. Mpumalanga, a rural province in the northeast of South Africa, implemented an enhanced surveillance system, which consisted of training hospital ICNs to rapidly report and correctly respond to nine infectious disease syndromes, including AFP. Weekly zero reporting is a component of the system. The non-polio AFP reporting rate per 100,000 children below the age of 15 years increased from 0.56 in 1997 to 0.91 in 1998 after introduction of the enhanced surveillance system, with more than 80% of the units reporting weekly. All units reported weekly from April 1999 to December 2001. Although non-polio AFP reporting rates were 0.27 (1999), 1.18 (2000) and 0.87 (2001), the 95% binomial exact confidence intervals for all years included 1 per 100,000. A review of paediatric admissions from January 1998 to December 2001 at all hospitals revealed that only five AFP cases had been missed by the enhanced surveillance system. The low international AFP reference rate and attendant variation expected due to chance, particularly in areas with relatively small populations, is an important factor that deserves more attention as we approach global polio eradication. PMID- 12753632 TI - Antibiotic therapy in murine filariasis (Litomosoides sigmodontis): comparative effects of doxycycline and rifampicin on Wolbachia and filarial viability. AB - The symbiosis of filarial nematodes and rickettsial Wolbachia endobacteria has been exploited as a target for antibiotic therapy of filariasis. Depletion of Wolbachia after tetracycline treatment results in filarial sterility because of interruption of embryogenesis and inhibits larval development and adult worm viability. The aim of this study was to investigate if antibiotic intervention of BALB/c mice infected with the rodent filaria Litomosoides sigmodontis with rifampicin or the combination of rifampicin and doxycycline can be used to shorten the treatment period. Both regimens, when given over a period of 14 days initiated with infection, were sufficient to deplete Wolbachia as evidenced by immunohistology and semiquantitative PCR. Worm development and filarial load were significantly reduced in experiments followed up until 63 days p.i. The therapy inhibited embryogenesis and led to filarial sterility. In contrast, treatment with doxycycline alone for 21 days led only to a modest reduction of Wolbachia, filarial growth retardation, worm viability and fertility. In conclusion, the combination of antirickettsial drugs could be used as a suitable tool to explore the minimum duration of therapy required for the depletion of Wolbachia in parasitized hosts subsequent to the onset of patency in human and animal filariasis and the prevention of adverse reactions in human infections. PMID- 12753633 TI - An estimation of the incidence of noma in north-west Nigeria. AB - Noma (cancrum oris, stomatitis gangrenosa) is a quickly spreading orofacial gangrene in children, caused by a combination of malnutrition, debilitation because of concomitant diseases (measles) and intraoral infections. The global incidence of noma in the world is uncertain. By comparing large numbers of noma patients and cleft lip patients in a large referral hospital for these disorders in Sokoto, Nigeria, we calculated the incidence of noma in north-west Nigeria as 6.4 per 1000 children. Extrapolation of this incidence to the developing countries bordering the Sahara Desert (the noma belt of the world) gives an incidence of 25,600 for that region and a global incidence of 30,000-40,000. Noma is a good biological parameter of extreme poverty, and hence a global monitoring system for noma can be justified. Though economic progress is the most effective preventive measure against noma, medical prevention by vaccination programmes against measles should be enhanced as well. PMID- 12753634 TI - Accuracy of perception and touch for detecting fever in adults: a hospital-based study from a rural, tertiary hospital in Central India. AB - OBJECTIVES: To evaluate the diagnostic accuracy of the patient's perception, and of the touch of patient attendants and a doctor for detecting fever. METHODS: We enrolled patients older than 13 years who presented with history of fever to the in- and out-patient departments of a rural teaching hospital. The design was a double-blind, cross-sectional analysis of a hospital-based case series, independently comparing reported history of fever and touch of patient attendant and that of doctor against an established reference standard (axillary temperature > 37.5 degrees C). Diagnostic accuracy was measured by computing sensitivity, specificity, and likelihood ratio values. The agreement between the patient, his attendant and the doctor was assessed by kappa statistic. RESULTS: We studied 462 patients of whom 274 (59.3%) were men. A total of 206 patients (44.58%) had fever. The patient's perception of fever (LR+ 1.77; 95% CI 1.52, 2.06), patient attendant's touch (LR+ 2.03; 95% CI 1.74, 2.36) and the doctor's touch (LR+ 3.08; 95% CI 2.51, 3.71) did not accurately distinguish those with and without fever. Doctors (LR- 0.20; 95% CI 0.17, 0.34) and patient attendants (LR- 0.24; 95% CI 0.14, 0.28) were more accurate in ruling out fever. The patient's perception agreed moderately with patient attendant's touch (kappa = 0.44; 95% CI 0.36, 0.53), and the doctor's assessment (kappa = 0.47; 95% CI 0.39, 0.55). There was moderate agreement between patients' attendants and the study doctor (kappa = 0.48; 95% CI 0.40, 0.56). CONCLUSIONS: Our findings suggest that patients, their attendants or doctors cannot accurately detect the presence of a fever without using a thermometer. Doctors should confirm a history of fever by recording temperature. PMID- 12753635 TI - Significance of Cryptosporidium as an aetiology of acute infectious diarrhoea in elderly Indians. AB - OBJECTIVE: To explore the significance of Cryptosporidium isolation in elderly Indian with acute infectious diarrhoea and its clinical correlates. DESIGN: A hospital-based clinico-aetiological study of 120 patients aged 60 years or older, 25 adults younger than 60 years and 25 children up to age 14 with acute diarrhoea, and 57 apparently healthy elderly individuals. RESULTS: Cryptosporidium was isolated in 22 older patients with diarrhoea (18.3%) and was highly significant (P < 0.01) compared with healthy age-matched controls. Of these patients 66% had a history of close contact with animals. Most (68%) Cryptosporidium infections occurred during the rainy season. Among the elderly patients 17% suffered from vomiting and abdominal pain, 31% were febrile; none were severely dehydrated. Stools numbered three to nine per day with duration of 5-17 days. Stool leucocytes were <6/hpf and no RBCs were seen. Isolation of Cryptosporidium in older persons was associated with diabetes mellitus (22.7%), tuberculosis (9.0%), malignancy (4.5%) and coronary artery disease (4.5%). CONCLUSION: Cryptosporidium is an important cause of diarrhoea in elderly Indians, especially those with close contact with animals. The infection has a mild clinical course, is self-limiting and does not cause dysenteric stool. Its effect on nutrition requires further study. PMID- 12753636 TI - Absence of the kdr mutation in the molecular 'M' form suggests different pyrethroid resistance mechanisms in the malaria vector mosquito Anopheles gambiae s.s. AB - Field tests conducted on adult Anopheles mosquitoes using standard WHO procedures, diagnostic kits and test papers in south-western Nigeria showed pyrethroid (deltamethrin and permethrin) resistance in adult populations of Anopheles gambiae sensu stricto. The knock-down resistance (kdr) mutation involved in pyrethroid resistance was only found in the molecular S form of An. gambiae s.s. even in area where both molecular M and S forms occurred in sympatry. The absence of the kdr mutation in the M form suggests an additional pyrethroid resistance mechanism in An. gambiae s.s. PMID- 12753637 TI - [Presence of Angiostrongylus cantonensis in Haiti]. AB - In the West Indies Angiostrongylus cantonensis was found first in Cuba in 1973, then in Puerto Rico (1986), in the Dominican Republic (1992) and recently in Jamaica (2002). Concurrently, the presence of the parasite was detected in the Bahamas and in New Orleans (LA, USA) in 1990. To assess the occurrence of A. cantonensis in Haiti, we investigated a number of rats in Port-au-Prince in 2002. Preliminary results among 23 captured and examined rats show that 75% (three of four) Rattus norvegicus and 21% (four of 19) R. rattus harboured A. cantonensis in their cardiopulmonary systems. Haiti is, for the first time, recognized as a new enzootic area for A. cantonensis in the Caribbean. This emerging zoonosis should be considered as a new public health hazard in Haiti. PMID- 12753638 TI - Treatment failure of pyrimethamine-sulphadoxine and induction of Plasmodium falciparum gametocytaemia in children in western Kenya. AB - Sub-Saharan Africa faces increasing levels of resistance of Plasmodium falciparum parasites to the first-line drug pyrimethamine-sulphadoxine (SP). Successful treatment with SP is reported to induce gametocytes and drug resistance may further increase gametocytaemia after treatment. Treatment success, gametocyte prevalence and gametocyte density were determined in 224 asymptomatic children in western Kenya on day 7 after treatment with SP. Treatment failure (R2 or R3 resistance) was observed in 22% of the children. The relative risk to show gametocytes on day 7 after treatment in children with treatment failure was 4.1 (95% CI 1.4-11.6) times higher compared to children with a sensitive infection, after adjustment for age and trophozoite density at the start of treatment. In addition, the gametocyte density was also higher upon SP treatment failure. These findings are reason for concern, as the increased gametocyte prevalence and density after SP treatment failure may increase the spread of SP-resistant strains in the population. PMID- 12753639 TI - Attitude of women in western Uganda towards pre-packed, unit-dosed malaria treatment for children. AB - In the context of a larger study on malaria related knowledge, attitudes, practices and beliefs in western Uganda 813 women aged 15-49 years were shown a sample of a pre-packed, unit-dosed malaria treatment for children, its use was explained and attitudes of the women were investigated. Of all women, 90.5% (86% urban, 92% rural) said they would prefer the pre-packed over the conventional type of treatment and 93.9% of these were willing to pay between 0.17 (rural) and 0.29 (urban) US dollars more for this treatment. Two-thirds (67.8%) thought that they would not have to ask their spouses before making a decision on the kind of treatment and 59.5% said they would rather stock the treatment at home than buy it when a child gets sick. The most mentioned reason for preferring pre-packs was their safety and cleanliness, while ease of application, dosing and compliance were secondary. We conclude that pre-packed, unit-dosed malaria treatment is accepted by the caretakers of children in the area studied and that they readily understand and accept its concept. This indicates a high potential for this approach to improve the home management of malaria fevers and reduce malaria related morbidity and mortality if adequate coverage can be achieved and if the intervention is embedded into an appropriate programme of behavioural change communication and provider training. PMID- 12753640 TI - The relationship between Anopheles gambiae density and rice cultivation in the savannah zone and forest zone of Cote d'Ivoire. AB - In 13 villages in the savannah zone and 21 villages in the forest zone of Cote d'Ivoire, the biting density of the principal malaria vector, Anopheles gambiae, was studied as a function of rice cultivation in the inland valleys in a 2-km radius around each village. In the savannah villages, during the main season cropping period, surface water on rice-cultivated and to a lesser extent on uncultivated inland valleys seems to contribute strongly to the A. gambiae population density. For the off-season cropping period (which starts after the first light rains in the savannah zone), correlations were weaker. Breeding sites other than in inland valleys may play an important role in the savannah zone. In the forest zone, however, the A. gambiae population density was strongly correlated with the surface water availability (SWA) in the rice-cultivated inland valleys, whereas the correlation with the SWA in other (uncultivated) inland valleys was weak. The requirement of sunlit breeding sites for A. gambiae might explain this difference between zones. In the forest zone, only inland valleys cleared for rice cultivation meet this requirement, whereas all other inland valleys are covered with dense vegetation. In the savannah zone, however, most undergrowth is burnt during the dry season, which permits sunlight to reach puddles resulting from the first rains. PMID- 12753642 TI - Living in the paddies: a social science perspective on how inland valley irrigated rice cultivation affects malaria in Northern Cote d'Ivoire. AB - The potential impact of irrigated agriculture on water-related vector-borne diseases has been an increasing source of concern for researchers from the bio medical sector. While most research on the potential impacts of irrigation on the health of local populations focuses on vector densities, levels of exposures, health services and technologies (prophylaxis, mosquito nets), we argue that it is essential to enlarge the scope of investigation and consider the complex mechanisms by which factors such as agriculture-generated changes in ecosystems, gender repositioning in the family organization as a result of access to new crops, and production activities combine together in increasing disease risks and producing new scenarios in the management of disease. This paper presents the results of an investigation of how transformations induced on the local society by the intensification of inland valley irrigated rice cultivation influence malaria health care systems and modulate risks to the health of local populations, within well-defined geographical boundaries in northern Cote d'Ivoire. Our results indicate that socio-economic transformations and gender repositioning induced, or facilitated, by the intensification of inland valley irrigated rice cultivation lead to a reduction of the capacity of women to manage disease episodes, contributing therefore to increase malaria incidence among farming populations. PMID- 12753641 TI - Inland valley rice production systems and malaria infection and disease in the savannah of Cote d'Ivoire. AB - In sub-Saharan Africa, lowlands developed for rice cultivation favour the development of Anopheles gambiae s. l. populations. However, the epidemiological impact is not clearly determined. The importance of malaria was compared in terms of prevalence and parasite density of infections as well as in terms of disease incidence between three agroecosystems: (i) uncultivated lowlands, 'R0', (ii) lowlands with one annual rice cultivation in the rainy season, 'R1' and (iii) developed lowlands with two annual rice cultivation cycles, 'R2'. We clinically monitored 2000 people of all age groups, selected randomly in each agroecosystem, for 40 days (in eight periods of five consecutive days scheduled every 6 weeks for 1 year). During each survey, a systematic blood sample was taken from every sick and asymptomatic person. The three agroecosystems presented a high endemic situation with a malaria transmission rate of 139-158 infective bites per person per year. The age-standardized annual malaria incidence reached 0.9 malaria episodes per person in R0, 0.6 in R1 and 0.8 in R2. Children from 0 to 9-year-old in R0 and R2 had two malarial attacks annually, but this was less in R1 (1.4 malaria episodes per child per year). Malaria incidence varied with season and agroecosystem. In parallel with transmission, a high malaria risk occurs temporarily at the beginning of the dry season in R2, but not in R0 and R1. Development of areas for rice cultivation does not modify the annual incidence of malarial attacks despite their seasonal influence on malaria risk. However, the lower malaria morbidity rate in R1 could be explained by socio-economic and cultural factors. PMID- 12753643 TI - Malaria and property accumulation in rice production systems in the savannah zone of Cote d'Ivoire. AB - Irrigation stabilizes agricultural production and hence improves farmers' living standards and conditions. The permanent presence of water may, however, increase the burden of water-related parasitic diseases and counter the economic benefits of irrigation by reducing farmers' health. The purpose of this study was to assess the impact of malaria on farm household property, beyond the health risk (studied elsewhere). The research question was: by weakening individuals, does malaria reduce productive capacities and income workers, and consequently limit their property accumulation? To test this hypothesis, we use data on property (farming equipment, livestock and durable consumer goods) and Plasmodium falciparum indicators generated by a study carried out in 1998 in the Ivorian savannah zone characterized by inland valley rice cultivation, with a sample of nearly 750 farming households. Property is influenced by many factors related to the size of the family, the area under cultivation and high parasite density infection rate of P. falciparum. A significant negative correlation between high density infection rate and the property values confirms that by reducing the living standards of households, malaria is a limiting factor for property accumulation. PMID- 12753645 TI - Unconventional myosins, actin dynamics and endocytosis: a menage a trois? AB - Ever since the discovery of class I myosins, the first nonmuscle myosins, about 30 years ago, the history of unconventional myosins has been linked to the organization and working of actin filaments. It slowly emerged from studies of class I myosins in lower eukaryotes that they are involved in mechanisms of endocytosis. Most interestingly, a flurry of recent findings assign a more active role to class I myosins in regulating the spatial and temporal organization of actin filament nucleation and elongation. The results highlight the multiple links between class I myosins and the major actin nucleator, the Arp2/3 complex, and its newly described activators. Two additional types of unconventional myosins, myosinIX, and Dictyostelium discoideum MyoM, have recently been tied to the signaling pathways controlling actin cytoskeleton remodeling. The present review surveys the links between these three classes of molecular motors and the complex cellular processes of endocytosis and actin dynamics, and concentrates on a working model accounting for the function of class I myosins via recruitment of the machinery responsible for actin nucleation and elongation. PMID- 12753646 TI - Unconventional motoring: an overview of the Kin C and Kin I kinesins. AB - All kinesins share a conserved core motor domain implying a common mechanism for generating force from ATP hydrolysis. How is it then that kinesins exhibit such divergent activities: motility, microtubule cross-linking and microtubule depolymerization? Although conventional motile kinesins have served as the paradigm for understanding kinesin function, the unconventional kinesins exploit variations on the motile theme to perform unexpected tasks. This review summarizes the biological functions and examines the possible molecular mechanisms of Kin C and Kin I unconventional kinesins. We also discuss the possible differences between the microtubule destabilization models proposed for Kar3 and Kin I kinesins. PMID- 12753647 TI - Clathrin- and dynamin-dependent coated vesicle formation from isolated plasma membranes. AB - We have developed a new rapid cell-free assay for endocytic clathrin-coated vesicle formation using highly purified rat liver plasma membrane sheets. After incubation in the presence of cytosol and nucleotides, released vesicles were collected by high-speed centrifugation and incorporated cargo receptors were detected by Western blotting. Three different cargo receptors were internalized into vesicles while a receptor, known to be excluded from coated pits, was not. The recruitment of cargo receptors into the vesicle fraction was cytosol, ATP and temperature-dependent and was enhanced by addition of GTP. Vesicle formation in this assay was confirmed by subcellular fractionation and EM analysis. Plasma membranes stripped of their endogenous coat proteins with 0.5 m Tris retained vesicle formation activity, which was highly dependent on clathrin and dynamin. Coat proteins and dynamin were not sufficient for clathrin-coated vesicle formation, and other peripheral membrane proteins recruited from the cytosol are required. The nonhydrolyzable ATP analogue, AMPPNP did not support clathrin coated vesicle formation; however, surprisingly, GTP gamma S was as effective as GTP. This assay will provide a powerful tool to dissect the minimum machinery and to probe the hierarchy of events involved in cargo selection and endocytic clathrin-coated vesicle formation. PMID- 12753648 TI - Non-classical export of an adenovirus structural protein. AB - The icosahedral capsids of Adenoviruses (Ads) consist of the hexon and stabilizing proteins building the facettes, and of the vertex protein penton base (Pb) anchoring the protruding fibers. The fibers bind to the Coxsackie virus B Ad cell surface receptor (CAR) and Pb to integrins. Here we describe a novel property of the Ad2 Pb. Pb was found to leave the infected cell and, upon exit, it attached to the surrounding noninfected cells forming a radial gradient with highest Pb levels on cells adjacent to the infected cell. The producer cells remained intact until at least 30 h post infection. At this point, Pb was not recovered from the extracellular medium, suggesting that its cell-cell spread might not involve free Pb. When viral particles were released at late stages of infection, soluble Pb was found in the extracellular medium and it randomly bound to noninfected cells. Nonlytic export of Pb occurred upon transient transfection with plasmid DNA, but plasmid-encoded fiber was not exported, indicating that cell-cell spread of Pb is autonomous of infection. Pb export was not affected by Brefeldin A-induced disruption of the Golgi apparatus, suggesting that it occurred via a nonclassical mechanism. Interestingly, the coexpression of Pb and fiber leads to both Pb and fiber export, termed 'protein abduction'. We suggest that fiber abduction might support viral dissemination in infected tissues by interfering with tissue integrity. PMID- 12753649 TI - Use of expression constructs to dissect the functional domains of the CHS/beige protein: identification of multiple phenotypes. AB - The Chediak-Higashi Syndrome (CHS) and the orthologous murine disorder beige are characterized at the cellular level by the presence of giant lysosomes. The CHS1/Beige protein is a 3787 amino acid protein of unknown function. To determine functional domains of the CHS1/Beige protein, we generated truncated constructs of the gene/protein. These truncated proteins were transiently expressed in Cos-7 or HeLa cells and their effect on membrane trafficking was examined. Beige is apparently a cytosolic protein, as are most transiently expressed truncated Beige constructs. Expression of the Beige construct FM (amino acids 1-2037) in wild type cells led to enlarged lysosomes. Similarly, expression of a 5.5-kb region (amino acids 2035-3787) of the carboxyl terminal of Beige (22B) also resulted in enlarged lysosomes. Expression of FM solely affected lysosome size, whereas expression of 22B led to alterations in lysosome size, changes in the Golgi and eventually cell death. The two constructs could be used to further dissect phenotypes resulting from loss of the Beige protein. CHS or beigej fibroblasts show an absence of nuclear staining using a monoclonal antibody directed against phosphatidylinositol 4,5 bisphosphate [PtdIns(4,5) P2]. Transformation of beige j fibroblasts with a YAC containing the full-length Beige gene resulted in the normalization of lysosome size and nuclear PtdIns(4,5)P2 staining. Expression of the carboxyl dominant negative construct 22B led to loss of nuclear PtdIns(4,5)P2 staining. Expression of the FM dominant negative clone did not alter nuclear PtdIns(4,5) P2 localization. These results suggest that the Beige protein interacts with at least two different partners and that the Beige protein affects cellular events, such as nuclear PtdIns(4,5)P2 localization, in addition to lysosome size. PMID- 12753650 TI - Endoplasmic reticulum retention, degradation, and aggregation of olfactory G protein coupled receptors. AB - The mammalian olfactory G-protein coupled receptor family is comprised of hundreds of proteins that mediate odorant binding and initiate signal transduction cascades leading to the sensation of smell. However, efforts to functionally express olfactory receptors and identify specific odorant ligand olfactory receptor interactions have been severely impeded by poor olfactory receptor surface expression in heterologous systems. Therefore, experiments were performed to elucidate the cellular mechanism(s) responsible for inefficient olfactory receptor cell surface expression. We determined that the mouse odorant receptors mI7 and mOREG are not selected for export from the ER and therefore are not detectable at the Golgi apparatus or plasma membrane. Specifically, olfactory receptors interact with the ER chaperone calnexin, are excluded from ER export sites, do not accumulate in ER-Golgi transport intermediates at 15 degrees C, and contain endoglycosidase H-sensitive oligosaccharides, consistent with olfactory receptor exclusion from post-ER compartments. A labile pool of ER-retained olfactory receptors are post-translationally modified by polyubiquitination and targeted for degradation by the proteasome. In addition, olfactory receptors are sequestered into ER aggregates that are degraded by autophagy. Collectively, these data demonstrate that poor surface expression of olfactory receptors in heterologous cells is attributable to a combination of ER retention due to inefficient folding and poor coupling to ER export machinery, aggregation, and degradation via both proteasomal and autophagic pathways. PMID- 12753652 TI - NKG2D ligands: unconventional MHC class I-like molecules exploited by viruses and cancer. AB - Our best teachers in revealing the importance of immune pathways are viruses and cancers that have subverted the most prominent pathways to escape from immune recognition. Viruses and cancer impair antigen presentation by classical MHC class I to escape adaptive immunity. The activating receptor NKG2D and its MHC class I-like ligands are other recently defined innate and adaptive immune pathways exploited by viruses and cancer. This review discusses recent advances in the understanding of how NKG2D, expressed on innate immune cells including natural killer cells, gammadelta+ T cells and macrophages, and adaptive immune cells such as CD8+ T cells, recognize stress-induced, MHC class I-like, self ligands. Moreover, we describe how viruses and cancer have developed strategies to evade this recognition pathway. PMID- 12753653 TI - HLA associations in type 1 diabetes: DPB1 alleles may act as markers of other HLA complex susceptibility genes. AB - Alleles at the HLA-DQB1, -DQA1 and -DRB1 loci are major determinants for susceptibility to develop type 1 diabetes (T1D). Increasing evidence supports that also other genes in, or near, the HLA complex contribute to the HLA-encoded risk. Alleles at the DPB1 locus have been suggested to directly influence the risk conferred by DQB1, DQA1 and DRB1 alleles, but the results are conflicting. We therefore genotyped 217 families from Norway, Denmark, Sweden and southern France to address the role of DPB1 alleles in T1D. After taking into account linkage disequilibrium (LD) with DQB1, DQA1 and DRB1 alleles, we found evidence that some DPB1 alleles are associated with modulating the risk of developing T1D. However, we show that the strong LD in the HLA complex, and the presence of extended haplotypes complicate the interpretation of the results. On DQ2-DR3 haplotypes, both allele 3 at microsatellite D6S2223 located 5.3-Mb telomeric of DPB1 and the extended DQ2-DR3-B18 haplotype display much stronger association than DPB1 alleles. When we exclude these effects, most of the apparent association of DPB1 alleles on DQ2-DR3 haplotypes disappear. Taken together, although we cannot completely rule out an effect of some DPB1 alleles, we propose that the statistically significant, albeit weak, DPB1 associations found are most likely the result of LD with another unidentified disease-susceptibility gene(s) in this region. PMID- 12753654 TI - IgG reactive to CTL-directed epitopes of self-antigens is either lacking or unbalanced in atopic dermatitis patients. AB - We previously demonstrated that CTL-directed epitopes derived from non-mutated self-antigens elicit a type-I allergy in the majority of healthy donors (HD) as did the presence of IgE and IgG reactive to these peptides in the sera of the donors. We investigated in this study whether Igs reactive to eight types of CTL directed peptides were elevated in the sera of 40 patients with atopic dermatitis (AD). Total IgE levels in the sera of AD patients were significantly higher than those of HD, however, no significant differences between the AD patients and the HD were observed in either the serum levels or the positive rates of IgE reactive to seven of the eight peptides. Total IgG levels were not different from each other, however, IgG reactive to the two peptides with no sequence similarity to other species and one peptide that had similarity to DNA helicase II of enterobacteria were not detectable in the sera of the AD patients. Although IgG reactive to the remaining five peptides, which had sequence similarity to other species, were detectable in both the AD patients and the HD, ratios of peptide specific IgG1/IgG2 were mostly lower in the AD patients than in the HD. These results indicate that IgG reactive to CTL-directed epitopes of self-antigens is either lacking or unbalanced in AD patients. This information may provide new insight into the immune-mechanisms of elevated auto-reactivity of AD patients. PMID- 12753655 TI - Alpha1 and alpha2 domains of Aotus MHC class I and Catarrhini MHC class Ia share similar characteristics. AB - Functional and structural analyses of major histocompatibility complex (MHC) class I molecules of the Aotus genus are necessary to validate it as a solid animal model for biomedical research. We thus isolated, cloned and sequenced exons 2 and 3 from three Aotus species (A. nancymaae, A. nigriceps and A. vociferans). We found 24 sequences, which divided into two different groups (Ao g1 and Ao-g2). A further sequence was identified as a processed pseudogene (Aona PS2). Both sequence evolution and variability analyses showed that Ao-g1 and Ao g2 display similar characteristics to Catarrhini's classical loci, such as positive selection pressure at the peptide binding region (PBR) high variability and a trans-specific evolution pattern. PMID- 12753656 TI - Association of Fcgamma receptor IIb and IIIb polymorphisms with susceptibility to systemic lupus erythematosus in Thais. AB - We recently reported association of a newly identified polymorphism of Fcgamma receptor (FcgammaR) IIb, I232T, with systemic lupus erythematosus (SLE) in Japanese. To date, information on FcgammaR genotypes and their association with SLE is limited in South-east Asian populations. To gain further insight into the role of FcgammaR polymorphisms in the genetic predisposition of SLE, association of FcgammaRIIa-H131R, IIb-I232T, IIIa-F176V and IIIb-NA1/NA2 (HNA-1a/1b) polymorphisms with SLE was analyzed in the Thai population, using case-control association analysis. FcgammaRIIb-232T/T and IIIb-NA2/NA2 genotypes were associated with SLE with the odds ratio of 2.55. Genotype relative risk analysis revealed significant association of IIb-232T/T and IIIb-NA2/NA2, and a tendency of association of the IIIa-176F/F genotype. Moreover, carriers of FcgammaRIIa 131R were significantly increased in patients with lupus nephritis. Significant linkage disequilibrium was present among FcgammaRIIb, IIIa and IIIb, and two locus analyses suggested that the tendency of association of FcgammaRIIIa could derive from linkage disequilibrium with IIb and IIIb. These results provided evidence that FcgammaR polymorphisms may be an important predisposing factor also in Thais in a complex manner. PMID- 12753658 TI - TNF-alpha polymorphisms and type 2 diabetes mellitus in Taiwanese patients. AB - Type 2 diabetic mellitus (type 2 DM) comprises more than 95% of all Taiwanese patients with DM. Tumor necrosis factor-alpha (TNF-alpha) expression is linked with insulin resistance, and is under strong genetic control. The correlation between TNF promoter genotypes and type 2 DM is still controversial, because discrepancies among different studies exist. Ethnic differences play certain roles in these conflicting results, because the distribution of TNF promoter polymorphisms is different among study subjects with different racial origins. Therefore, we examined the relationship between the incidence of type 2 diabetes in Taiwanese and two polymorphisms of the TNF-alpha promoter region (positions 238 and -308) as well as the correlation between these polymorphisms and the patients' biochemical manifestations. Genomic DNA was extracted from peripheral blood cells of 261 Taiwanese patients with type 2 DM and 189 non-diabetic control study subjects, and their TNF promoter G-238A and G-308A polymorphisms were analyzed by PCR-RFLP analysis. No significant association between TNF-alpha G 238A and G-308A polymorphisms with type 2 diabetic incidence was observed. However, associations between TNF-alpha G-238A and low-density lipoprotein cholesterol and between G-308A promoter polymorphism and high-fasting plasma glucose levels, using multiple linear regression analysis with adjustment for the subjects' age, sex, body mass index and diabetic status, were found. Our results suggested that though TNF-alpha G-238A and G-308A polymorphisms were not involved in the pathogenesis of type 2 DM, type 2 diabetic patients carrying TNFA-A or TNF 308*2 genotype might be more susceptible to diabetic complications such as atherosclerosis. PMID- 12753657 TI - HLA alleles in isolated populations from North Spain: origin of the Basques and the ancient Iberians. AB - HLA-A, -B, -DRB1, -DQA1 and -DQB1 alleles have been studied in three relatively isolated populations of northern Spain from Cantabria ( Pas Valleys inhabitants or Pasiegos and Cabuernigos) and from the Basque Country (Arratia Valley inhabitants). These populations have been compared with neighbouring ones and other Mediterraneans by using neighbour-joining dendrograms and plane genetic distances. PMID- 12753659 TI - Identification of three new DRB1 alleles, DRB1*0107, *0425 and *13012 and confirmation of DRB4*01033. AB - The characterization of three novel DRB1 alleles is described, DRB1*0107, DRB1*0425 and DRB1*13012 as well as confirmation of DRB4*01033. Two alleles, DRB1*0107 and *0425, showed amino acid differences with previously identified HLA molecules. In DRB1*0107, the glutamine at position 10 was substituted by a glutamic acid. DRB1*0425 showed one amino acid difference with DRB1*0418 (I to F) at position 67, and five amino acid differences with DRB1*04011 at positions 67 (L to F), 70 (Q to D), 71 (K to R), 74 (A to L) and 86 (G to V). The alleles DRB1*13012 and DRB4*01033 had protein sequences identical to DRB1*13011 and DRB4*01031/01032, respectively. Nucleotide differences were present at position 306 for DRB1*13012 and at position 321 for DRB4*01033. PMID- 12753660 TI - New allele frequency database: http://www.allelefrequencies.net. AB - A new website (http://www.allelefrequencies.net) has been compiled to execute the frequency of alleles at various polymorphic regions of different populations in the field of histocompatibility and immunogenetics. Data for HLA alleles has been added but this will be extended to include frequency data of polymorphisms in other immunogenetic regions, e.g. cytokines, KIR receptors, MIC. PMID- 12753661 TI - Nomenclature for factors of the HLA system, update October 2002. PMID- 12753662 TI - Nomenclature for factors of the HLA system, update November 2002. PMID- 12753663 TI - Nomenclature for factors of the HLA system, update December 2002. PMID- 12753664 TI - HFE, the MHC and hemochromatosis: paradigm for an extended function for MHC class I. AB - HFE was discovered as the hereditary hemochromatosis (HH) gene. It is located on chromosome 6 (6p21.3), 4Mb telomeric to the HLA-A locus, and its product has a structure similar to MHC class I molecules. HFE encodes two frequent mutations: C282Y and H63D. One of these (C282Y) is present in a large proportion of Caucasian HH patients. HFE has a tissue distribution compatible with a role in iron absorption (intestine), recycling (macrophages) and transport to the fetus (placenta). PMID- 12753665 TI - Allelic variation of the inducible costimulator (ICOS) gene: detection of polymorphisms, analysis of the promoter region, and extended haplotype estimation. AB - The human chromosome region 2q33 including the three costimulatory molecules CD28, CTLA-4 and ICOS, has been subject to much attention due to its linkage to a number of autoimmune diseases. The search for the causal relationship of this linkage has revealed several polymorphisms, but no variations in the amino acid sequences, except for one polymorphism in the leader sequence of CTLA-4. In the present study, we examined the ICOS gene of an unrelated group of healthy donors from the Danish population. We were able to report 16 intronic SNP, one intronic G-insert and two repeat regions in intron 4, consistent with the [T]n and the [GT]n regions reported in a Japanese study. Putative haplotypes for the established SNP and repeat polymorphisms have been estimated by computational analysis. Sequencing of approximately 3500 bp of the upstream region of ICOS revealed an additional eight SNP of which two resided in putative NF-kB and Sp1 sites. In accordance with previous studies we detected no variations in the coding regions except for a rare polymorphism that was found in one donor in the last codon of exon 5, which lead to a heterozygous genotype, but no amino acid change. This suggests that regulation of transcription rather than protein structure could be a possible mechanism in the explanation of linkage. PMID- 12753666 TI - Distribution of HLA-A alleles in eight ethnic groups from Pakistan. AB - The extreme polymorphism found at some loci of the HLA system has made it an invaluable tool for population genetic analyses. In this study eight diverse ethnic groups from Pakistan were analyzed at the HLA-A locus using sequence specific primers for polymerase chain reaction (PCR-SSP) and then further typed to the allele level using a two-stage sequence specific oligonucleotide probe (SSOP) strategy. Four of these ethnic groups (Burusho, Hazara, Kalash, Pathan) were from the north and four (Baloch, Brahui, Sindhi and Parsi) were from the south of Pakistan. Nine alleles were identified as unique to a particular ethnic group within Pakistan. Maximum variation was seen in the HLA-A*02 allele family for which 11 alleles were detected in the eight Pakistani ethnic groups. The alleles that showed significant variation between the Pakistani ethnic groups include A*0101, A*0206, A*0209, A*0207, A*0217, A*1101, A*2402/09 N/11 N, A*2902, A*3301 and A*3001. A phylogenetic tree based on DA distances for HLA-A allele frequencies separated the Pakistani populations from other world populations and also separated the only Dravidian speaking population of Pakistan, the Brahui, from the remaining Indo-European speaking ethnic groups of Pakistan. PMID- 12753667 TI - Genetic affinities among Mongol ethnic groups and their relationship to Turks. AB - The central Asian country Mongolia is home to more than 20 tribes and ethnic groups, some of which are related to neighboring Turkic populations. The main Mongolian people, Khalkha, live in central and eastern Mongolia while the Tsaatan minority lives in the north of the country. The Oold minority is from the western Altai mountain region and live in close proximity with Turkic people. We have typed the HLA-A, -B, -Cw, -DRB1 and -DQB1 loci by PCR-SSP in these three Mongolian populations as well as a sample of the German population. To examine their genetic relationships, a sample of the Turkish population already typed at the HLA-A, -B and -DRB1 loci were used. Altogether five populations were analyzed: Khalkha (n = 100), Tsaatan (n = 72), Oold (n = 52), German (n = 260) and (Anatolian) Turkish (n = 498). Nei's unbiased genetic identity (GI) and genetic distance (GD) were estimated from genotypes using PopGene v1.31, and dendrograms were constructed using phylip. The results suggested a close relationship of the Khalkha to the Tsaatan. The Turks and Germans were equally distant to all three Mongolian populations. These results confirmed the lack of strong genetic relationship between the Mongols and the Turks despite the close relationship of their languages (Altaic group) and shared historical neighborhood. This study has provided useful population data for genetic and anthropologic studies bridging eastern and western populations. PMID- 12753668 TI - Hyperkeratosis and leukocytosis in transgenic mice carrying MHC class I chain related gene B (MICB). AB - Major histocompatibility complex (MHC) class I chain-related gene A and B (MICA and MICB) are located very close to HLA-B. MICA is reported to be strongly associated with Behcet's disease (BD), a multisysytemic inflammation disorder characterized by oral apthous ulcers, skin lesions and genital ulcers. These two molecules are highly conserved at the amino acid levels. To determine the function of MICB in vivo and the relationship between the expression of MICB and BD experimentally, we produced several transgenic mouse lines (termed CAG-MICB) expressing human MICB cDNA under a ubiquitous promoter. They exhibited a 50% increase in the number of white blood cells compared with their non-transgenic littermates, and also exhibited a 10-20% reduction in body weight compared with non-transgenic littermates. Exfoliation of the skin first appeared around 7 days after birth and disappeared after 2 weeks of age. This was repeatedly observed in the transgenic offspring of two independent CAG-MICB lines examined. Histopathological analysis of skin of young mice exhibiting skin abnormalities revealed hyperkeratosis of the epidermis and thickening of the granular layer with slight infiltration of inflammatory cells in the dermis without any vasculitis. Other remarkable abnormalities associated with BD have not been observed in the CAG-MICB lines. Furthermore, fluorescein angiography of eyes of the CAG-MICB lines was performed, but there were no marked changes of BD-related uveitis in the ocular fundus. These findings suggest that (i) MICB expression is related to temporary skin inflammation, and (ii) expression of MICB is not directly associated with BD. PMID- 12753669 TI - Additional factor in some HLA DR3/DQ2 haplotypes confers a fourfold increased genetic risk of celiac disease. AB - Although HLA-DQ genes are the major celiac disease (CD) susceptibility genes, results from Finnish families suggest that not all DQ2-encoding haplotypes confer equal susceptibility to CD, implying the effect of other gene(s) in the HLA region. The aim of the present work was to extend and confirm the aforementioned results in a southern European population ( Italian) and to better localize the additional risk factor/s. The association of nine loci spanning the HLA region from DR to HFE, 4.5-Mb telomeric of HLA-A, was tested. The analysis was performed by comparing marker frequencies in DR3-DQ2 haplotypes transmitted and non transmitted to the affected offspring in 156 Italian CD families selected for having at least one DR3-positive parent. The same analysis was performed independently in 101 Finnish CD families selected with the same criteria. Three alleles, MICA-A5.1, MICB-CA24 and MIB-350, all characteristic of the B8-DR3 extended haplotype, showed a significantly increased frequency in DR3 transmitted haplotypes in the Italian families. DR3 haplotypes carrying the combination of these alleles conferred an approximate fourfold increased CD risk. B8-DR3 transmitted haplotypes were significantly more conserved telomerically down to the MIC-Class I region. Similar results were seen in the Finnish families. The major conclusion that holds true in both populations is that, while DQ2 is an absolute requirement for the development of CD, the presence of an additional genetic factor within the MIC-Class I region confers an approximate 4-fold increased risk of the disease. PMID- 12753671 TI - Immunogenetics of two new HLA alleles: A*0108 and B*4031. AB - Two new alleles, HLA-A*0108 and B*4031, were identified in north-western European Caucasoid subjects. A*0108 differed from A*010101 by a single substitution (C to T) at position 216 in exon 3, resulting in an amino acid difference of Arg to Trp at position 163. It was present on a haplotype with B*1501/60/70/71; Cw*0303; DRB1*1301; DRB3*0202; DQA1*0103; DQB1*0603 and its product reacted as a normal HLA-A1 specificity. B*4031 differed from B*4001 by two nucleotides in exon 3 (positions 20 (G to C) and 69 (A to G)) resulting in two amino acid differences (Arg to Ser at position 97 and Asn to Asp at position 114). It was found on a haplotype with HLA-A*03; Cw*0304; DRB1*0404/32; DRB4*0101/3/5; DQA1*03; DQB1*0302 and has the HLA-B60 specificity. Both alleles have frequencies of < 0.0002 in the largely north-western European Caucasoid blood donor population resident in Wales. PMID- 12753670 TI - A single round PCR method for genotyping human surfactant protein (SP)-A1, SP-A2 and SP-D gene alleles. AB - The genes coding for the human surfactant proteins (SP)-A and SP-D are located on chromosome 10q22-q23.1. SP-D is the product of a single gene whereas SP-A is the product of two highly homologous genes SP-A1 and SP-A2. Several single nucleotide polymorphisms (SNP) are present in the SP-A1, SP-A2 and SP-D genes. Because of this high degree of sequence homology between the SP-A1 and SP-A2 genes, current genetic analysis studies employ a nested PCR/radioactive hybridization or restriction fragment length polymorphism approach to initially isolate the genes and subsequently to detect the SNP in these isolates. In this manuscript, we report the primers and conditions of a sequence specific primer-PCR methodology that enables the identification of SP-A1, SP-A2 and SP-D gene allelic variants directly on genomic DNA material. PMID- 12753672 TI - Expression defect of an HLA-A*24 allele associated with DNA methylation in a normal individual. AB - Non-expressed HLA alleles become a potential problem in the transition of the HLA typing methodology from serologic typing to more accurate DNA typing. In this study, a novel nonexpressed A*24 allele identified from two members of a Korean family was characterized. At the DNA sequence level, the nonexpressed allele (A*24023) is apparently normal; the complete genomic sequence was identical to HLA-A*2402101, from the 5'-upstream region to the 3'-downstream region, except for a single silent substitution at codon 211 (GCG-->GCA) in exon 4. A DNA methylation analysis using methylation-sensitive restriction enzymes, however, showed that the nonexpressed A*24023 allele from an apparently normal individual was highly methylated in regions covering exons and introns as well as the 5' upstream region. This result suggests that hypermethylation of the HLA-A gene may induce gene inactivation in the normal individuals. PMID- 12753673 TI - Observations on the HLA-A2403 specificity. AB - We found HLA-A*2423 in a subject possessing the HLA-A2403 specificity. Consequently we delineated the A*24 alleles of 41 subjects previously assigned as 'A2403' by serology and/or 'low-resolution' PCR-SSP (allele group: A*2403/10/18/22/23/33) and found that 36 were A*2403, 3 were A*2410 and 2 were A*2433. The A*2410 and A*2433 subjects also typed as A2403. Several likely A*2410/23/33-bearing haplotypes were identified and the frequency of the A*2403/10/18/22/23/33 alleles was determined in 26,826 Welsh blood donors. We suggest that HLA-A9/A24 antibodies against epitope(s) involving 166D and 167G will fail to react with all A*2403/10/18/22/23/33 products. As it is this 'short' A9/A24 serological reactivity that defines HLA-A2403 all these products will be serologically designated as HLA-A2403. PMID- 12753674 TI - Are HMG-CoA reductase inhibitors underutilized in dialysis patients? AB - Patients with end-stage renal disease (ESRD) treated with dialysis have a dramatically elevated rate of cardiovascular disease (CVD) compared to the general population. Lipid-lowering therapy with 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors ("statins") has been shown to markedly reduce cardiovascular risk in patients without renal failure, but their effect has not been fully studied in the dialysis population. In this article we will first discuss the known benefits of statin therapy in the general population and summarize the current guidelines for such therapy. We will then examine the evidence linking dyslipidemia and cardiac disease in the dialysis population and discuss possible pathophysiologic mechanisms by which statins could prevent cardiac disease in these patients. We will also review prior clinical studies of the effects of statins in patients on dialysis, with particular attention to the safety and efficacy of these drugs in this population. Finally, we will review how statins are currently being used in the care of dialysis patients and suggest whether an expanded utilization of these drugs could help reduce the enormously high rates of cardiac disease in this patient population. PMID- 12753675 TI - Hidden sources of phosphorus in the typical American diet: does it matter in nephrology? AB - Elevated serum phosphorus is a major, preventable etiologic factor associated with the increased cardiovascular morbidity and mortality of dialysis patients. An important determinant of serum phosphorus is the dietary intake of this mineral; this makes dietary restriction of phosphorus a cornerstone for the prevention and treatment of hyperphosphatemia. The average daily dietary intake of phosphorus is about 1550 mg for males and 1000 mg for females. In general, foods high in protein are also high in phosphorus. These figures, however, are changing as phosphates are currently being added to a large number of processed foods including meats, cheeses, dressings, beverages, and bakery products. As a result, and depending on the food choices, such additives may increase the phosphorus intake by as a much as 1 g/day. Moreover, nutrient composition tables usually do not include the phosphorus from these additives, resulting in an underestimate of the dietary intake of phosphorus in our patients. Our goal is to convey an understanding of the phosphorus content of the current American diet to better equip nephrologists in their attempt to control hyperphosphatemia. PMID- 12753676 TI - Overcoming barriers to arteriovenous fistula creation and use. AB - National guidelines advocate the placement of arteriovenous fistulas (AVFs) as the preferred vascular access for hemodialysis (HD) patients because of their low complication rate, lower costs, and prolonged patency, once matured. The current Dialysis Outcomes Quality Initiative (DOQI) guidelines aim for an AVF incidence of 50% and a 40% prevalence in the United States. Although patients currently starting dialysis do so at an increasingly older age and with more comorbidity, they should be given every opportunity to receive an AVF. Meeting this challenge is facilitated by a multidisciplinary approach with early referral to the nephrologist in the predialysis period for access planning. Key components of a vascular access program may include the coordination by a dedicated access coordinator and outcome tracking via a prospective database. Preoperative vessel evaluation and careful selection of an appropriate surgical site, along with an experienced surgeon, improve surgical outcomes. Transposed brachiobasilic or other tertiary fistulas should be offered to patients who cannot receive a native radiocephalic or brachiocephalic fistula. The ability to routinely monitor and salvage failing AVFs is important to achieving successful AVF outcomes. Standardized definitions of AVF outcomes are important to allow individual centers and continuous quality assurance (CQA) programs to track and benchmark their outcomes against local and national standards to help them meet recommended targets. PMID- 12753677 TI - The challenges of diabetes care in the dialysis unit. AB - Nephrologists, dialysis facilities, and payers are confronted with a new and more difficult set of challenges to effectively care for the steadily increasing number of patients with diabetes mellitus (DM) developing end-stage renal disease (ESRD). U.S. Renal Data System (USRDS) data suggest that the current care of patients with DM on dialysis is suboptimal. Recently published reports have confirmed the value of HbA1C measurements in the diabetic dialysis population, that control of blood glucose lowers mortality, and that a program of care management and diabetes education can have a significant impact on patient outcomes. As leader of the nephrology team, the nephrologist should, at a minimum, be accountable for defining who is managing the diabetes. A more systematic and educated approach to DM and its complications needs to be developed by the renal community. PMID- 12753678 TI - A time for rediscovery: chronic hemofiltration for end-stage renal disease. AB - Provision of maintenance renal replacement therapy (MRRT) is becoming increasingly challenging. The number of patients requiring MRRT is growing rapidly, while the number of nephrologists, nurses, and other staff available to provide therapy is not increasing at a similar rate. Patients are now older and have more comorbid conditions, which magnifies the complexity of their RRT and makes it increasingly difficult to maintain the quality of care within the limitations of constrained economic resources. The two most commonly used MRRTs are hemodialysis (HD) and peritoneal dialysis (PD). A third modality of MRRT, hemofiltration, is not currently in widespread use in North America. In hemofiltration, plasma is filtered through a highly permeable, biocompatible synthetic membrane and waste products are removed by convection and solvent drag as the filtrate moves across the membrane. Hemofiltration equipment constantly monitors the rate of plasma removal and administers a sterile, nonpyrogenic replacement solution at an appropriate rate to replace the waste-laden ultrafiltrate and avoid vdume depletion in the patient. Hemofiltration may offer some potential clinical advantages to the MRRT patient, including better hemodynamic and cardiovascular stability, enhanced removal of middle molecular weight toxins, increased comfort, and therapy preference. Some reports indicate possible reduced morbidity and mortality and reduced risk of bacterial contamination and inflammation as a result of the use of sterile, ultrapure replacement fluids. Chronic hemofiltration is a relatively simple MRRT option that may offer significant benefits for many patients who have end-stage renal disease (ESRD) and deserves closer consideration for these patients. PMID- 12753679 TI - Systolic hypertension in hemodialysis patients. AB - Systolic hypertension with or without diastolic hypertension is a major problem in hemodialysis (HD) patients; isolated diastolic hypertension is uncommon. Accelerated age-related changes in vascular stiffness, together with factors peculiar to uremia, lead to loss of large and small vessel distensibility and profound changes in circulatory function that includes an increase in systolic pressure and widening of the pulse pressure. Epidemiologic studies show a direct relationship of mortality with systolic blood pressure (BP) and an inverse relationship with diastolic BP. Thus systolic BP should be the focus of treatment. In HD patients with systolic hypertension, diastolic BP is inversely related to cardiovascular risk. An accurate diagnosis of hypertension followed by nonpharmacologic measures (sodium restriction, exercise, dry weight) should be the initial steps in BP reduction. The second step should be the use of antihypertensive agents, particularly the use of angiotensin converting enzyme (ACE) inhibitors and/or beta-blockers. The use of these agents has been associated with better outcomes in observational studies in HD patients. Furthermore, the administration of atenolol and lisinopril can be supervised three times a week to achieve improved BP control. Daily dialysis may improve BP and cardiovascular risk factors. Although more difficult to implement, it may emerge as a feasible alternative to conventional dialysis. Adequate systolic BP control with these available and emerging techniques should help stem the tide of cardiovascular mortality and mortality in HD patients. PMID- 12753680 TI - How can the use of arteriovenous fistulas be increased? PMID- 12753685 TI - Uremic malnutrition: new insights into an old problem. AB - Uremic malnutrition is highly prevalent and is associated with poor clinical outcomes in end-stage renal disease (ESRD) patients. Inadequate diet and a state of persistent catabolism play major roles in predisposing these patients to uremic malnutrition and appear to have an additive effect on overall outcome. Recent studies highlight the existence of a complex syndrome involving chronic inflammation, metabolic abnormalities, and hormonal derangements contributing to the increased morbidity and mortality observed in ESRD patients. Novel strategies such as appetite stimulants, anti-inflammatory drugs, and anabolic hormones along with conventional nutritional supplementation may provide potential interventions to improve clinical outcome in ESRD patients. PMID- 12753686 TI - Human immunodeficiency virus infection in end-stage renal disease patients. AB - Human immunodeficiency virus (HIV) infection in patients with end-stage renal disease (ESRD) offers many diagnostic and therapeutic challenges to nephrologists. Renal failure may be a direct consequence of viral infection (HIV associated nephropathy), or intrinsic renal diseases may occur in previously infected individuals. Patients receiving renal replacement therapy (RRT) may acquire HIV infection from blood transfusions, renal allografts, sexual contacts, or needle sharing by drug addicts. In the early 1980s, the overall prognosis of patients with the acquired immunodeficiency syndrome (AIDS) was very poor, and survival of those with ESRD was dismal. Consequently many even questioned the value of providing maintenance dialysis to patients with AIDS. With advances in diagnostic techniques in serologic and viral markers of disease, and deployment of highly effective antiretroviral agents, the prognosis of HIV-infected patients has dramatically improved. Over the past two decades, experiences in the management of HIV patients with ESRD is accumulating. Both peritoneal dialysis and hemodialysis are effective modes of therapy and many centers are now beginning to perform renal transplantation in HIV-infected patients. This article deals with various aspects of HIV infection in patients with ESRD. PMID- 12753688 TI - Complications of percutaneous treatment of thrombosed hemodialysis access grafts. AB - Percutaneous catheter-based thrombolysis is commonly used in association with angioplasty to treat thrombosed hemodialysis arteriovenous grafts. Although major complications of these percutaneous procedures are relatively uncommon, they can result in several potentially serious complications, including pulmonary embolism, cerebral embolism, arterial embolism, bleeding with perigraft hematoma or hemorrhage, and vein rupture. This article reviews the epidemiology, clinical significance, and management of these complications. PMID- 12753687 TI - Thrombosis in end-stage renal disease. AB - Although renal failure has classically been associated with a bleeding tendency, thrombotic events are common among patients with end-stage renal disease (ESRD). A variety of thrombosis-favoring hematologic alterations have been demonstrated in these patients. In addition, "nontraditional" risk factors for thrombosis, such as hyperhomocysteinemia, endothelial dysfunction, inflammation, and malnutrition, are present in a significant proportion of chronic dialysis patients. Hemodialysis (HD) vascular access thrombosis, ischemic heart disease, and renal allograft thrombosis are well-recognized complications in these patients. Deep venous thrombosis and pulmonary embolism are viewed as rare in chronic dialysis patients, but recent studies suggest that this perception should be reconsidered. Several ESRD treatment factors such as recombinant erythropoietin (EPO) administration, dialyzer bioincompatibility, and calcineurin inhibitor administration may have prothrombotic effects. In this article we review the pathogenesis and clinical manifestations of thrombosis in ESRD and evaluate the evidence that chronic renal failure or its management predisposes to thrombotic events. PMID- 12753689 TI - Hemodialysis catheter-associated endocarditis: clinical features, risks, and costs. AB - Endocarditis associated with vascular access catheters for hemodialysis (HD) is a catastrophic but not widely appreciated phenomenon. Its current incidence, clinical outcome, and associated costs are not easily ascertained. Increasing use of tunneled catheters for HD access may result in a larger pool of patients at risk for endocarditis. We present two representative cases, review recent trends, and assess the current potential for additional cases. PMID- 12753690 TI - Peritoneal dialysis underutilization: the impact of an interventional nephrology peritoneal dialysis access program. AB - Peritoneal dialysis (PD) is an underutilized form of renal replacement therapy. Recent data have emphasized that only 12% of end-stage renal disease (ESRD) patients are initiated on this form of therapy in the United States. Patients requiring PD have most often been referred to general surgeons for catheter placement. This has incurred additional delays in starting treatment and loss of decision-making control by the referring nephrologist. To address this issue, we developed and incorporated our own PD access placement program into the preexisting chronic kidney disease (CKD) education program. To date, 46 patients have undergone 71 procedures. These included 51 (72%) PD catheter insertions, 14 (20%) removals, and 6 (8%) repositioning procedures for poor drainage. PD catheter insertion was performed peritoneoscopically under local anesthesia and a Fogarty catheter was used to reposition a migrated catheter. All of the procedures were performed by nephrologists in a dedicated interventional nephrology (IN) laboratory. All six repositioning procedures failed to restore optimal drainage. Five of these patients had the catheter removed and a new catheter placed during the same procedure. Of these five patients, one had recurrence of poor drainage and opted for hemodialysis (HD). The sixth patient declined reinsertion and chose HD. Of the remaining seven removal procedures, three were due to fungal peritonitis, one due to bowel perforation, one due to severe depression, one due to transplant, and one catheter was removed at the request of the primary physician in a terminally ill patient. Eight of the 51 catheter insertions were during the initial admission of a catastrophic dialysis start. Two of these patients started acute PD and avoided catheter placement for HD. Thirty-seven of 46 patients have a functional PD catheter with a follow-up of 8.6 +/- 0.8 (mean +/- SE) months. During an 18-month period our PD population has increased from 43 to 80 patients. We conclude that a dedicated PD access placement program coupled with a CKD education program can have a dramatic impact on patient choice and PD growth. PMID- 12753691 TI - Pharmacomechanical thrombolysis of natural vein fistulas: reduced dose of TPA and long-term follow-up. AB - Twenty-five episodes of pharmacomechanical thrombolysis of 20 clotted arteriovenous fistulas (AVFs) are reported. The technique presented utilizes the local instillation of tissue plasminogen activator (TPA) in small doses together with manual maceration to dissolve clot and balloon angioplasty to correct the underlying stenoses. Since the minimum dose of TPA necessary to successfully perform thrombolysis of a natural vein fistula had never been determined, an attempt to use as minimal a dose of TPA as possible was made. Five procedures were performed in fistulas which had previously undergone a thrombolysis procedure with TPA. The procedures were successful in 92% of cases with an average dose of TPA required of 2.3 +/- 0.32 mg/procedure. In addition to the 20 accesses in this article, we offer follow-up life table data on 15 fistulas that were previously reported for a total of 35 accesses salvaged with pharmacomechanical thrombolysis. Primary patency was 11.2 months and secondary patency was 25 months. Fifty-five percent of fistulas required repeat angioplasty procedures at an average of 3.6-month intervals. In addition, more than half of the fistulas that presented with clotting required repeat interventions for continued patency. This report demonstrates the effectiveness of small doses of TPA in successful pharmacomechanical thrombolysis of clotted fistulas. PMID- 12753692 TI - Nephrogenic fibrosing dermopathy: an unusual skin condition associated with kidney disease. AB - We report the case of a patient who, while on long-term hemodialysis (HD), developed nephrogenic fibrosing dermopathy, a newly described sclerosing skin disorder. This disorder is characterized by thickened, hardened skin with brawny hyperpigmentation and raised plaques. The most common patient complaints are pruritus and dysesthesia. The extremities are predominantly involved with sparing of the torso and face. Dysfunction of internal organs has not been described, distinguishing it from other fibrosing conditions such as scleroderma. The skin biopsy is characterized by haphazardly arranged dermal collagen spindle cells in the reticular dermis. Extensive mucin deposits are interposed between collagen bundles and there are an increased number of fibroblast-type cells. In contrast to scleroderma, inflammatory cells are generally absent. Corticosteroid therapy can be tried, but in our patient was of no benefit. PMID- 12753693 TI - An elevated hemidiaphragm 3 months after internal jugular vein hemodialysis catheter placement. AB - Phrenic nerve palsy following central venous catheterization is a rare complication and is not well recognized. We present a 33 months old girl who has renal failure secondary to nephrotic syndrome. A left internal jugular catheter was placed using the Seldinger technique after a single injection of 2 ml prilocaine hydrochloride for local anesthesia and a single internal jugular vein cannulation. Subsequent chest roentgenograms confirmed proper catheter and diaphragm position. Three months after catheter placement, decreased breath sounds on the left side of the chest were noted. Left phrenic nerve palsy was demonstrated with fluoroscopy and electromyography with external diaphragmatic electrodes. The nerve damage was delayed after catheter placement, it seems unlikely that it was related to direct nerve trauma from the cannulation needle, local anesthetic infiltration of the nerve, or subsequent hematoma formation in this case. The phrenic nerve is in close proximity to both the catheter and the vein in which the catheter rests, an inflammatory reaction related to the catheter has been suggested as the cause for the nerve damage. PMID- 12753694 TI - Positive PPD, history of BCG, and dialysis. PMID- 12753695 TI - Skin thickness and subcutaneous erythropoietin. PMID- 12753696 TI - Safety of high doses of non-dextran irons. PMID- 12753699 TI - Approaches to quality improvement in nursing homes: lessons learned from the six state pilot of CMS's Nursing Home Quality Initiative. AB - BACKGROUND: In November 2002, the Centers for Medicare & Medicaid Services (CMS) launched a Nursing Home Quality Initiative that included publicly reporting a set of Quality Measures for all nursing homes in the country, and providing quality improvement assistance to nursing homes nationwide. A pilot of this initiative occurred in six states for six months prior to the launch. METHODS: Review and analysis of the lessons learned from the six Quality Improvement Organizations (QIOs) that led quality improvement efforts in nursing homes from the six pilot states. RESULTS: QIOs in the six pilot states found several key outcomes of the Nursing Home Quality Initiative that help to maximize the potential of public reporting to leverage effective improvement in nursing home quality of care. First, public reporting focuses the attention of all stakeholders in the nursing home industry on achieving good quality outcomes on a defined set of measures, and creates an incentive for partnership formation. Second, publicly reported quality measures motivate nursing home providers to improve in certain key clinical areas, and in particular to seek out new ways of changing processes of care, such as engaging physicians and the medical director more directly. Third, the lessons learned by QIOs in the pilot of this Initiative indicate that certain approaches to providing quality improvement assistance are key to guiding nursing home providers' desire and enthusiasm to improve towards a using a systematic approach to quality improvement. CONCLUSION: The Nursing Home Quality Initiative has already demonstrated the potential of public reporting to foster collaboration and coordination among nursing home stakeholders and to heighten interest of nursing homes in quality improvement techniques. The lessons learned from this pilot project have implications for any organizations or individuals planning quality improvement projects in the nursing home setting. PMID- 12753700 TI - [From genomics, proteomics to cytomics, or from cytometry to cytomics]. AB - Cytomics, an ad hoc term, represents a new field which connects cyto- with omics. The cyto- arises from analytical cytology or cytometry, which has developed for more than three decades and includes a serial of quantitative technology which can separate the heterogenetic cell populations into different cytomes by molecular probes. The -omics comes from proteomics which is determined by genomics. Therefore, cytomics is to separate the cytomes by cytometry technology and identify their proteomes controlled by genome or special genes. Cytomics project consists of two parts of technology, one is separation of cytomes and another is post-sorting analysis. Development of cytomics related technology would help us to understand molecular basis of different cytomes and the transition between them, to gradually grasp the details of genomics, and to know of how to apply them for clinical work. Thus, a new horizon will appear. PMID- 12753698 TI - Effects of Greek Orthodox Christian Church fasting on serum lipids and obesity. AB - BACKGROUND: No study to date has focused on the impact of Greek Orthodox Christian fasting on serum lipoproteins and obesity yet. METHODS: 120 Greek adults were followed longitudinally for one year. Sixty fasted regularly in all fasting periods (fasters) and 60 did not fast at all (controls). The three major fasting periods under study were: Christmas (40 days), Lent (48 days) and Assumption (August, 15 days). A total of 6 measurements were made during one year including pre- and end-fasting blood collection, serum lipoprotein analyses and anthropometric measurements. RESULTS: Statistically significant end-fasting total and LDL cholesterol differences were found in fasters. Fasters compared to controls presented 12.5% lower end-total cholesterol (p < 0.001), 15.9% lower end LDL cholesterol (p < 0.001) and 1.5% lower end-BMI (p < 0.001). The end- LDL/HDL ratio was lower in fasters (6.5%, p < 0.05) while the change in end- HDL cholesterol in fasters (4.6% decline) was not significant. Similar results were found when the pre- and end-fasting values of fasters were compared. No change was found in control subjects. CONCLUSIONS: Adherence to Greek Orthodox fasting periods contributes to a reduction in the blood lipid profile including a non significant reduction in HDL cholesterol and possible impact on obesity. PMID- 12753701 TI - [Detailed mapping and clinical significance of loss of heterozygosity on 9p13-23 in laryngeal squamous cell carcinoma by microsatellite analysis]. AB - BACKGROUND & OBJECTIVE: Microsatellites are the repeated DNA sequences scattered widely within the biological genomes and closely linked with many important genes. In carcinogenesis, microsatellites often display loss of heterozygosity(LOH) as tumor suppressor genes. Some microsatellite loci often exist in the hot spots of LOH at high frequency in some specific maligances. The tumor suppressor genes, which are associated with the development and progression of the tumor, possibly harbor in the vicinity of these hot spots. Therefore, the study of LOH by microsatellite analysis is an important way to detect the putative tumor suppressor genes. This study was designed to refine the hot spots of LOH on 9p13-23 in laryngeal squamous cell carcinoma and compare the correlation between the incidence of microsatellite LOH and the clinicopathological parameters. METHODS: Tumor tissues were obtained from paraffin embedded sections with microdissection. Genomic DNA was extracted from tumor tissues and peripheral blood lymphocytes with the phenol-chloroform. Polymerase chain reaction(PCR) amplification and denaturing gel electrophoresis were performed on a set of 42 laryngeal squamous cell carcinoma and corresponding peripheral blood lymphocytes using 13 highly polymorphic microsatellite markers on 9p13-23. The correlation was analyzed between microsatellite LOH at the high frequency on 9p13-23 and clinicopathological characteristics in the patients with squamous cell carcinoma of larynx. RESULTS: (1)Of the 42 laryngeal cancers, 41(97.6%) showed LOH in at least one of the microsatellite markers tested on 9p13 23. The most frequently deleted marker was D9S162 in 17 of the 19 (89.5%) informative samples. The marker D9S171, which is located on 9p21, had LOH detected in 12 of the 15 informative cases (80.0%). LOH at the D9S1748 marker (closest to the p16 gene locus) was detected in 18 of the 36 informative cases (50.0%). (2)Allelic deletion mapping revealed two minimal regions of LOH encompassing markers D9S161-D9S171 on 9p21 and IFNA-D9S162 on 9p22-23. (3) Multiple LOH (>or= 4) on 9p21-23 was found more frequently in the patients under 60 years, with supraglottic squamous cell carcinoma or cervical lymph node metastasis than those over 60 years, with glottic squamous cell carcinoma or without cervical lymph node metastasis (P< 0.01,P< 0.01,P< 0.05, respectively). On the contrary, there was no correlation between T stages or pathologic classification and the frequency of LOH on 9p21-23 in 42 squamous cell carcinoma of larynx. CONCLUSION: These findings imply the presence of at least two putative tumor suppressor genes on 9p13-23 in laryngeal squamous cell carcinoma. Multiple genetic alterations are probably implicated in supraglottic squamous cell carcinoma with cervical lymph node metastasis in younger patients. PMID- 12753702 TI - Ribozyme targeted on HPV16E6 mRNA induced apoptosis on human cervical carcinoma CaSKi cells. AB - BACKGROUND & OBJECTIVE: Human papillomavirus is related to cervical cancer. Ribozyme is special kind of RNA that can cleave target RNA. The aim of this study was to investigate the characterization of the cultured cervical cancer cell line transfected with anti-HPV16E6-ribozyme (HRz) and the effect of ribozyme on proliferation and apoptosis of cervical cancer cell. METHODS: Ribozyme targeted on HPV16E6 mRNA was designed using computer. With the method of lipofectin transfection, the anti-HPV16E6- ribozyme and empty eucaryotic expressing plasmids were transfected into the CaSKi cells, which named as CaSKi-R and CaSKi-P, respectively. The expression of E6 mRNA in the three kinds of cells was examined by Northern blot analysis. Cell cycle was determined using flow cytometry. Cell apoptosis rate was examined using fluorescent (Hoechst) staining and TUNEL (TDT mediated dUTP nick end labeling). The expression of certain proteins, including HPV16E6, c-myc, bcl-2, p53, and Fas, were also determined using flow cytometry. RESULTS: RNA dot blot analysis demonstrated that HRz mRNA expressed stably in the CaSKi-R cells. Northern blot analysis showed that the expression of E6 mRNA was much lower in the CaSKi-R cells than that in the CaSKi cells, while there was no difference of E6 mRNA levels between the CaSKi cells and the CaSKi-P cells. The apoptosis rate in the CaSKi-R cells was much higher than that in the CaSKi cells and the CaSKi-P cells. The cell cycle was arrested in G2 phase, with decrease in percentage of S phase cells. Anti-HPV16E6-ribozyme can significantly reduce the expression of E6, c-myc, and bcl-2 genes in the CaSKi-R cells, and increase the expression of p53 compared with that in Caski cells. This phenomenon was not found in the CaSKi-P cells. The expression of Fas was similar in the three kinds of cells. CONCLUSION: Ribozyme targeted on HPVE6 mRNA can induce apoptosis of human cervical cancer CaSKi cells. The reason may be the decrease of expression of E6 gene, and the successive changes of expression of some genes, including c myc, bcl-2, and p53 genes. PMID- 12753704 TI - [Survivin mutants reverse the malignancy of HeLa cells]. AB - BACKGROUND & OBJECTIVE: Survivin was aberrantly expressed in most cancer tissues, suggesting that survivin plays an important role in carcinogenesis. This study was designed to investigate the function and mechanism of survivin mutants in tumor cells. METHODS: The site-mutant and truncated survivin mutants were transfected into HeLa cells and selected using G418. Cell apoptosis was analyzed using flow cytometry. Protein level of cyclin D1 was detected by Western blot analysis. RESULTS: Survivin mutant plasmid expressed in the HeLa cells successfully. The expressed protein could be detected using related antibody. Colony formation ability significantly decreased in the HeLa cells with survivin mutants compared with that in the parental HeLa cells. The HeLa cells transfected instantly with survivin mutants could undergo apoptosis automatically. Meanwhile, survivin mutants could cause an increase of multinuclear HeLa cells. The effect of survivin-N showed more effective than that of survivin T34A. Survivin-N and survivin T34A could influence the expression of cyclin D1 and reduced its protein levels of 68% and 12%, respectively. CONCLUSION: Survivin mutants can partially reverse the malignancy of HeLa cells. The reduction of cyclin D1 induced by survivin mutants may play an important role in it. Survivin may be a target gene in gene therapy of cancer. PMID- 12753703 TI - [Proteomics-based identification of Maspin differential expression in bronchial epithelial immortalized cells and malignant transformation cells]. AB - BACKGROUND & OBJECTIVE: Maspin, a serepin inhibitor, plays a key role in tumor growth and metastasis. The aim of this study was to identify the differential expression of Maspin in malignant transformation process of bronchial epithelial cells by proteomics. METHODS: Functional proteomics analysis of Maspin on bronchial epithelial immortalized cells and malignant transformation cells was carried out using immobilized pH gradient (IPG) two-dimensional electrophoresis, peptide mass fingerprinting (PMF), and post source decay (PSD) of bio-mass spectrometry. RESULTS: Nearly 1500 expressed proteins profile on bronchial epithelial immortalized cells and malignant transformation cells were obtained in the range of MW 14.4-94 kDa, PI 3-10. Image analysis showed that Maspin was down regulated in malignant transformation cells compared with that in immortalized cells. Northern blot analysis showed that the mRNA abundance of Maspin in malignant transformation cells was much lower than that in immortalized cells. CONCLUSION: Alteration expression of Maspin at transcription and translation levels might be involved in carcinogenesis of lung. PMID- 12753705 TI - [Expression of apoptosis and proliferation related genes in carcinogenesis of lung squamous cell carcinoma in rats]. AB - BACKGROUND & OBJECTIVE: The activation of caspase-3 protein, located the downstream of apoptosis, is the key of cell apoptosis signal conduction. Caspase 3 is the most important performer in accelerating apoptosis in the caspase family. Much progress has been achieved in the study of caspase-3 and human non small cell lung cancer. This study was designed to investigate the effect of cellular proliferation and apoptosis related genes caspase-3 and proliferating cell nuclear antigen (PCNA), and to seek whether they could be chosen as molecular biology markers for lung cancer. METHODS: 3-Methylcholanthrene (MCA) and diethyinitrosamine (DEN) were used to induce lung squamous cell carcinoma by intra-left lobar-bronchial instillation in 50 Wistar rats. The other 10 rats instilled with iodized oil were regarded as control group. The expression of caspase-3 and PCNA were evaluated by SP immunohistochemistry during carcinogenesis. TUNEL(TDT-mediated dUTP nick end labeling) method was used to examine apoptotic cells. RESULTS: For the rat bronchial epithelium cells in control group, precancerous lesions, and lung squamous cell carcinoma, positive coefficient values of caspase-3 protein were 3.10+/-0.99, 2.25+/-1.13, and 1.38+/ 0.95 on average, respectively; the means of PCNA-labeling index (PCNA-LI)were 14.10+/-5.02, 28.13+/-8.72, and 41.88+/-14.24, respectively; the means of apoptotic index(AI) were 0.60+/-0.52, 2.06+/-0.85, and 2.26+/-1.14, respectively.Significant differences in caspase-3 protein expression were observed between bronchial epithelium in control group and lung squamous cell carcinoma (P< 0.01). Caspase-3 expression was showed stronger in precancerous lesions than that in lung cancer (P< 0.05). Low proliferation index and low AI were detected in rat bronchial epithelium region in control group,which were significant differences in precancerous lesions and lung cancer, respectively (P< 0.01). In 34 rats with lung squamous cell carcinoma, there was negative relationship between caspase-3 and PCNA-LI (r=-0.7306, P< 0.01), so did it in AI and PCNA-LI(r=-0.8127,P< 0.01), but there was no relationship between caspase-3 expression and AI(P >0.05). CONCLUSION: Loss of caspase-3 expression may be associated with the development of lung squamous cell carcinoma in Wistar rats, but it is not associated with AI. PCNA-LI is an important marker for malignant progression of lung cancer. PMID- 12753706 TI - [Analysis of splicing variants in NASG 3'UTR, down-regulated in nasopharyngeal carcinoma, and its expression in multiple cancer tissues]. AB - BACKGROUND & OBJECTIVE: NASG gene, a tissue-specific gene of human nasopharyngeal epithelium was isolated by suppression subtractive hybridization. This study was designed to analyze splicing variants in NASG 3'untranslated region (UTR) and its expression profiling in multiple cancer tissues. METHODS: The PCR primers were designed in NASG 3'UTR around the splicing variants and reverse transcription polymerase chain reaction (RT-PCR) was performed. The PCR products were separated and sequenced. The expression patterns of NASG were detected by RT-PCR among nasopharyngeal carcinoma (NPC) cell line HNE1, primary human embryo nasopharyngeal epithelial cells, NPC biopsies, and normal adult nasopharyngeal epithelial tissues. Its expression profiling in multiple cancer tissues were tested by cancer profiling array hybridization. RESULTS: There were three splicing variants in NASG 3'UTR. NASG was identified to be down-regulated in NPC cell line HNE1 and 71% of the NPC biopsies, but up-regulated in 25% lung of the cancer biopsies, and not express in other cancer tissues and normal tissues. CONCLUSION: There were three splicing variants in NASG 3'UTR. Its abnormal expression may be an important molecular event in NPC and lung cancer. PMID- 12753707 TI - [Effects of Epstein-Barr virus latent membrane protein 1(EBV-LMP1) on related factors of metastasis of nasopharyngeal carcinoma cell line CNE1]. AB - BACKGROUND & OBJECTIVE: It has been proved that Epstein- Barr virus (EBV) latent membrane protein 1 (EBV-LMP1) can induce the expression of matrix metalloproteinase-9 (MMP-9). This study was designed to investigate the effect of EBV-LMP1 on related factors of metastasis of nasopharyngeal carcinoma cell line CNE1. METHODS: Expression of MMP-9 was studied in human NPC cell lines cultured in vitro: CNE1 (well differentiated cell line of NPC) and CNE1-GL (CNE1 cell line transfected with an eukaryotic LMP1-expression plasmid) by SP immunohistochemistry and Western blot analysis. Cell-matrix adhesion assay was used to study the adhesive ability of CNE1-GL cells. The effects of LMP1 on the invasion and migration of CNE1 cells were investigated by transwell methods. RESULTS: MMP-9 was expressed in both cell lines but the intensity of the staining was different. The positive rates of expression of MMP-9 in CNE1 and CNE1-GL cells were 30.2% and 98.2%, respectively (P< 0.05). The increased expression of MMP-9 was also shown in CNE1-GL cells by Western blot analysis. Cell-matrix adhesion assay showed that the adhesive ability of CNE1-GL with the matrix (mean A value: 1.2508+/-0.0711) was higher than that of CNE1 cell (mean A value: 0.9519+/-0.068) (P< 0.001). Invasion assay and migration assay showed that the invasion and migration of CNE1-GL cell were higher than those of CNE1 cells (P< 0.01). CONCLUSION: The transfection of LMP1 can increase the expression of MMP-9 in CNE1 cells. Abilities of adhesion, migration, and invasion of CNE1 cell were induced by LMP1. It is suggested that MMP-9 may have a role in the LMP1-induced acceleration of invasion and metastasis of NPC cells. PMID- 12753708 TI - [Relationship between human telomerase reverse transcriptase transcriptional level and telomerase activity in three ovarian cancer cell lines]. AB - BACKGROUND & OBJECTIVE: Approximately 90% of tumors have telomerase activity, whereas most normal cells do not express telomerase. Telomerase catalytic subunit or human telomerase reverse transcriptase (hTERT) is the main component of telomerase. Telomerase expression is predominantly regulated at the transcriptional level of hTERT. Telomerase, hTERT, and hTERT promoter are closely related. The aim of this study was to investigate the relationship among human telomerase reverse transcriptase (hTERT) promoter activity, hTERT mRNA expression, and telomerase activity in three ovarian cancer cell lines. METHODS: hTERT promoter activity was determined by luciferase assay after the plasmids of pBTdel-279 containing hTERT core promoter were transfected into three ovarian carcinoma-derived cell lines of OVCAR3, SKOV3, 3AO and normal human ovarian epithelial cells. hTERT mRNA expression levels of these cells were determined by semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR). Telomerase activity was determined by PCR-ELISA (enzyme-linked immunosorbent assay) in above four cells. Immortalized human embryonic kidney cell line HEK293 and the human embryonic lung fibroblasts HELF were used as positive control and negative control, respectively. RESULTS: The relative luciferase activities of OVCAR3, SKOV3, 3AO and normal ovarian epithelial cells by the hTERT promoter were 31.4%, 20.3%, 17.7%, and 0.3%, respectively, as the luciferase activity of pGL3 control plasmid in each cell line was considered as 100%. The hTERT mRNA relative expression levels of above four cells were 1.30, 1.00, 0.63, and 0, respectively, as SKOV3 expression level was considered as 1. The telomerase activities were 0.580, 0.414, 0.386, and 0.103, respectively, as >0.2 was considered as positive. The hTERT promoter activity, hTERT mRNA expression level, and telomerase activity were specially raised in three ovarian cancer cell lines. The highest levels of hTERT promoter activity, hTERT mRNA expression, and telomerase activity were observed in OVCAR3 cell line, whereas negative in normal ovarian epithelial cells. hTERT promoter activity was closely associated with hTERT mRNA expression level and telomerase activity (P< 0.01,P< 0.02). CONCLUSION: hTERT promoter is specially activated in ovarian cancer cells which expresses telomerase. hTERT promoter activity is positively correlated with telomerase activity. Acting as a targeting promoter, hTERT promoter may be applied in gene therapy of ovarian cancer. PMID- 12753710 TI - [Expression and significance of MRP, GST-pi, Topo IIalpha, and LRP in gastric carcinoma]. AB - BACKGROUND & OBJECTIVE: Multidrug resistance-associated protein (MRP), glutathione-S-transferase-pi(GST-pi), topoisomerase IIalpha(Topo IIalpha)and lung resistance protein (LRP) play important roles in the multidrug resistance(MDR)of tumor chemotherapy. There were few reports on combined determination of the expression of MRP, GST-pi, Topo IIalpha and LRP in gastric carcinoma. This study was designed to investigate the expression and significance of MRP, GST-pi, Topo IIalpha and LRP in gastric carcinomas. METHODS: Immunohistochemistry SP method was used to determine the expression of MRP, GST-pi, Topo IIalpha, and LRP in 90 tumor samples from the patients with gastric carcinoma. Chi-square test and Fisher exact test were used to analyze the significance of the expression. RESULTS: (1)The positive expression rates of MRP, GST-pi, Topo IIalpha and LRP in gastric carcinoma were 88.9%, 91.1%, 74.4%, and 87.7%, respectively. They were all significantly higher than those in normal stomach tissues (P< 0.05). (2)The expression levels of MRP, GST-pi, and LRP in well-moderated differentiation adenocarcinoma were significantly higher than those in poor differentiation adenocarcinoma. The expression of Topo IIin well-moderated differentiation adenocarcinoma was significantly lower than that in poor differentiation adenocarcinoma. There was no difference between the expression levels of them in different degree of invasion or with lymph nodes metastasis and without lymph nodes metastasis (P > 0.05). (3) There was no correlation in any two items among the expression levels of MRP,GST-pi, topo IIalpha,and LRP. CONCLUSION: MRP, GST pi, topo IIalpha,and LRP play important roles in the primary MDR of gastric carcinoma. The expression of them are associated with the differentiation, but are not associated with the invasion degree and lymph node metastasis. PMID- 12753709 TI - [HPV detection and FHIT expression in esophageal squamous carcinoma from high incidence area in Cixian County]. AB - BACKGROUND & OBJECTIVE: Cixian County is one of the high incidence areas of esophageal carcinoma in China. Up to now, no work has been conducted on the possible etiological role of human papilloma virus (HPV) infection on esophageal carcinoma in this local area. The aim of this study was to explore the putative roles of HPV infection on the esophageal carcinogenesis of the patients in Cixian and to elucidate the possible relationship between HPV existence and fragile histidine triad gene (FHIT gene, a putative tumor suppressor gene), expression in squamous cell carcinoma of esophagus. METHODS: The existence of HPV DNA and the expression of FHIT gene at protein level in esophageal carcinoma tissues were determined with PCR and immunohistochemical staining, respectively, in 128 archival paraffin-embedded tissue blocks of esophageal squamous carcinoma from the high incidence area of Cixian and 24 tissue blocks from the non-high incidence area. RESULTS: PCR results showed that the positive detection rate of HPV in esophageal carcinoma tissues from the high incidence area was 20.3%, which was slightly higher than that from the non-high incidence area (8.3%)(P >0.05). Immunohistochemically, aberrant expression of FHIT gene in esophageal carcinoma tissue was found in 75.6% cases from the high incidence area and only 54.2% from the non-high incidence area, which was significantly lower than that of the former (P< 0.05). No correlation was found between the abnormal expression of FHIT and existence of HPV DNA in esophageal squamous carcinoma tissues. CONCLUSION: HPV DNA could be detected in partial esophageal carcinoma cases from the high incidence area of esophageal carcinoma in Cixian. The aberrant expression rate of FHIT protein in the cases from the high incidence area of esophageal carcinoma in Cixian is higher than that from the non-high incidence area. PMID- 12753711 TI - [Experimental study of c-jun protein expression on proliferation and apoptosis of laryngeal cancer cells]. AB - BACKGROUND & OBJECTIVE: c-jun is an early protooncogene and its protein product is a component of activating protein-1 (AP-1) transcription factor. c-jun participates in the transcription of many growth factors and cytokines. Recently it has been found that the expression level and activity of c-jun protein in tumor cells abnormally increased and c-jun protein controlled the proliferation, surviving, and apoptosis of tumor cells. In this study, the expression of c-jun protein in laryngeal cancer and its relationship with clinical statistics were discussed to evaluate the controlling role of c-jun protein in laryngeal cancer. METHODS: Samples from Department of Otolaryngology, The First Affiliated Hospital of China Medical University, were acquired from May 2001 to February 2002. These samples including 52 laryngeal cancer tissues, 15 vocal polyps, and 10 normal laryngeal tissues were investigated individually by immunohistochemistry to observe the localization and expression of c-jun protein in these three kinds of samples. RESULTS: The expression of c-jun protein in laryngeal cancer (56.41+/ 24.8%) was not only significantly higher than that in vocal cord (32.48+/-1.78%) and laryngeal tissue (no expression), but also was correlated with the differentiating degree of laryngeal cancer tissue and lymph node metastasis(P< 0.01); however, the expression of c-jun was not associated with clinical staging(P >0.05). CONCLUSION: The expression level of c-jun protein significantly increased in laryngeal cancer cell and its expression may be considered as an indicator for differentiating degree,neck lymph node metastasis, and prognosis of laryngeal cancer. PMID- 12753712 TI - [Effect of cell cycle on telomerase activity of hepatoma cells and its relationship with replication of hepatitis B virus]. AB - BACKGROUND & OBJECTIVE: Previous studies have shown that the expression of telomerase activity is closely correlated with the formation and development of tumor cells. Furthermore, the cell cycle is associated with hepatitis B virus (HBV) replication level and telomerase activity. For further study their relationship, this experiment was designed to investigate the effect of serum deprivation or all-trans-retinoic acid(RA)on cell cycle of human hepatoma cells transfected by HBV DNA (HepG2 cell line) and the associations of cell cycle with telomerase activity and HBV replication. METHODS: Human hepatoma HepG2 cells were respectively treated with serum deprivation or RA. Cell cycle was analyzed using flow cytometry. Telomerase activity was determined quantitatively by TRAP-PCR ELISA. HBV-DNA in culture media was determined using quantitative PCR and semiquantitative dot blot hybridization assay. HBsAg and HBeAg in cell culture media were measured using quantitative ELISA. RESULTS: RA treatment or serum deprivation inhibited the proliferation of HepG2 cells and the cells were arrested at G(0)/G(1) phase. The percentages of G(0)/G(1) phase of RA group and serum deprivation were 68.3% and 65.2%, respectively, while that of control group was 43.1% (P< 0.01). The levels of telomerase activity also significantly decreased. The absorbance values that represented the telomerase activity of RA group and serum deprivation group were 0.32 and 0.41, respectively, while that of control group was 1.34(P< 0.01). In addition,HBV replication of HepG2 cells remarkably increased, which was shown as high products of HBV-DNA, HBsAg and HBeAg in culture media of RA group and serum deprivation group. The contents of HBV DNAs were 4.4x10(6), 5.1x10(6), and 1.2x10(6) copies/ml in RA group, serum deprivation group, and control group, respectively(P< 0.01). The values of P/N of HBsAg were 3.5, 3.7, and 1.3 in RA group, serum deprivation group, and control group, respectively (P< 0.01). The values of P/N of HBeAg were 19.8, 22.5, and 13.4 in RA group, serum deprivation group, and control group, respectively (P< 0.01). CONCLUSION: Telomerase expression was associated with cell cycle in HepG2 cells. Telomerase was mainly expressed in S phase of cell cycle. HBV replication was also closely correlated with cell cycle, which increased in quiescent hepatocytes and decreased in proliferating hepatocytes. PMID- 12753713 TI - [Operative treatment of pheochromocytoma: report of 11 cases]. AB - BACKGROUND & OBJECTIVE: Pheochromocytoma is clinically uncommon. Pheochromocytoma patients without hypertension or symptoms are often misdiagnosed. The patients could be cured by surgical removal of the tumor. However, surgery is dangerous if the preoperative preparation was not well done. The authors reported their experiences in surgical treatment of pheochromocytoma. METHODS: Clinical data of 11 patients with pheochromocytoma were reviewed retrospectively to summarize the management of blood pressure during pre-operation and operation. RESULTS: After excision of the tumors, 11 patients have been followed up for 3 months to 10 years. Blood pressure became normal and symptoms vanished in all patients. Only one patient recurred at paraveterbral sympathetic chain 6 months after operation and underwent another operation for removal of the tumor. CONCLUSION: Surgery is the mainstay of therapy for pheochromocytoma. The adequate preoperative preparation is very important for successful surgery. PMID- 12753714 TI - [Analysis of the results of 143 cases of pituitary micro-adenoma treated by Linac X-Knife stereotactic radioneurosurgery]. AB - BACKGROUND & OBJECTIVE: The trans-sphenoid surgery was one of the main methods for the treatment of functional pituitary micro-adenoma for many years, while the development of radiosurgical techniques in recent years has given neurosurgeons new ways to treat functional pituitary micro-adenoma. This report aimed to ascertain the effectiveness of X-Knife stereotactic radioneurosurgery on pituitary micro-adenoma. METHODS: From June 1996 to June 2001, 143 cases of functional pituitary micro-adenoma were treated by X-Knife radiosurgery, in which 73 cases of secreting prolactin (PRL),54 cases of secreting growth hormone(GH), 13 cases of secreting adrenal corticotrophin hormone (ACTH), and 3 cases of secreting both PRL and GH. The radiosurgical procedure included local anesthesia administration, head-ring mounting, serial thin slice CT scanning with contrast enhancement, CT image transfer, treatment planning on the computer workstation, stereotactic localization of the target onto the Linac isocenter, and then the Linac was used to perform multiple non-coplannar arc radiation. All cases received single fraction radiosurgery. RESULTS: After X-Knife radiosurgery, follow-up ranged 3-60 months (median 38.4 month). Among those 112 cases that acquired follow-up, 78(69.7%) cases showed good clinical improvements, 10(8.9%) cases showed relatively good clinical improvements, 19(16.8%) cases showed no clinical improvement, and 5(4.6%) cases had progression of disease and enlargement of the adenoma. CONCLUSION: X-Knife radiosurgery is a safe and effective technique for the treatment of functional pituitary micro-adenoma. While for young patients, the diagnosis of pituitary micro-adenoma and use of X Knife radiosurgery should be very cautious, otherwise hypopituitarism might be resulted. PMID- 12753715 TI - [Value of manifestations of magnetic resonance imaging (MRI) in diagnosis of parotid tumors and their pathological bases]. AB - BACKGROUND & OBJECTIVE: The pathologic types of parotid tumor are very complicated and the advantage of magnetic resonance imaging (MRI) in diagnosis of these tumors is very obvious; however, there is not still accordance at several areas of MRI diagnosis of them. A large amount of cases are needed for analyzing these MRI features and their pathological fundament. This study was conducted to summarize the MRI manifestations of common benign and malignant tumors and to explore the value of the morphological features of MRI in histological diagnosis of parotid tumor. METHODS: The MRI manifestations for 132 cases(140 lesions) of parotid tumor/lesion were collected, including 89 benign tumors/lesions and 43 malignant tumors. Of them, 112 cases were proved by surgery, 17 cases by biopsy, and 3 cases by clinic observation. MRI features were completely compared with pathological results of surgical samples slice by slice in 58 cases. T(1)WI and T(2)WI sequence were used in MRI unenhancement scan; 108 cases (115 lesions) were examined with both unenhancement and enhancement. Chi-square test was used for statistic analysis. RESULTS: (1) The MRI features of common parotid benign tumors were as following: 40 cases of Warthin's tumors enhanced slightly and more than half of them (25 cases) appeared low or equal signal intensity in T(2)WI; 22 cases of pleomorphic adenoma appeared high and unhomogeneous signal intensity in T(2)WI; 5 cases of hemangioma were characterized by their vascular structures which were enlarged than normal; 3 cases of lymphangioma not only appeared specially irregular shape but also surrounded adjacent structures of parotid gland (3 cases). The features of 4 cases of parotid lipoma were same as those in other position of the body.(2) The MRI features of common parotid malignant tumors were as following: 8 cases of malignant pleomorphic adenoma appeared high and unhomogeneous signals in T(2)WI together with irregular shape and ill-defined margin; 7 cases of adenoid cystic carcinoma characterized by invasion of very large range surrounding parotid gland. In 8 cases of malignant mucoepidermoid carcinoma, 3 cases showed low-grade malignant mucoepidermoid carcinomas, which had well-defined margin and were difficult to distinguish from benign tumor. However, 5 cases of high-grade malignant mucoepidermoid carcinoma showed ill defined margin, their features were tend to undergo necrosis and invade lymph node; 9 cases of lymphomas were all secondary lesions and there were two characters in their imaging, one was the wide involved range, the other was they consisted of several nodes. The shapes of 3 cases acinic cell carcinoma could be either regular or irregular, the cystic areas could be found in them with high signal intensity both on T(1)WI and T(2)WI. CONCLUSION: There are certain characters for each of common benign and malignant parotid tumors in MRI and these may be helpful for histological diagnosis in most of them. PMID- 12753716 TI - [Effect of intraoperative and postoperative radiotherapy and chemotherapy for intermediate and advanced pancreatic head carcinoma]. AB - BACKGROUND & OBJECTIVE: The excision rate is low for the intermediate and advanced pancreatic head carcinoma. Comprehensive treatment is the main method to prolong patient's life. The objective of this study was to discuss the effect of intraoperative and postoperative radiotherapy on intermediate and advanced pancreatic head carcinoma. METHODS: Twenty-seven patients with intermediate or advanced pancreatic head carcinoma underwent cholecystojejunostomy combined by intraoperative radiotherapy with direct electron beam on the tumor. Meanwhile the catheter of multifunctional implantable drug delivery system was inserted via gastroduodenal artery for postoperative perfusion chemotherapy. The survival rate was calculated using the direct method after follow-up for 3-29 months. RESULTS: All carcinoma shrank with 100% pain releasing rate. The 6-, 12-,and 24-month survival rates were 100%, 93.7%, and 20%, respectively. Five patients died and the average survival period was 17.9 months. CONCLUSION: Intraoperative radiotherapy combined with perfusion chemotherapy is an effective method to prolong the life of the patients with intermediate or advanced pancreatic head carcinoma. PMID- 12753717 TI - [Malignant fibrous histiocytoma of head and neck: clinical analysis of 21 cases]. AB - BACKGROUND & OBJECTIVE: Malignant fibrous histiocytoma (MFH) is a type of pleomorphic neoplastic diseases with complex pathological structure. Its histological origin is uncertain. It was often classified as other carcinoma. This study was designed to investigate the clinical and pathological features and improve the diagnosis and treatment. METHODS: To summarize and analyze the clinical experiences of 21 cases of MFH at head and neck proved histologically from June 1984 to June 1999 treated in Department of Head and Neck Surgery, Henan Tumor Hospital. RESULTS: Twenty cases were followed up more than 3 years; one case was lost. The 3-year survival rate was 42.9%(9/21). Nineteen cases were treated with surgery. Two cases in advanced stage were treated by non-surgery who died in 2 and 5 months. Among the patients treated with surgery, 6 cases survived without evidence of recurrence more than 3 years, 13 cases recurred within 2 years and 9 cases with metastasis. Seven cases received second surgery after recurrence. Among them,3 cases survived more than 3 years after second surgery. Of all 21 patients, 12 were proved with cervical lymph node metastasis. CONCLUSION: MFH at head and neck region is a kind of malignant disease with high recurrent rate and the cervical lymph node metastasis rate was 57.1%. Amplified radical surgery is the first choice of treatment. The second surgery has special value to the recurrent patients. Radiotherapy alone or chemotherapy alone is not effective to MFH of head and neck region. PMID- 12753718 TI - [Relationship between expression of multiple tumor suppressor(MTS1) and E cadherin and metastasis of breast carcinoma]. AB - BACKGROUND & OBJECTIVE: Most of breast cancer patients died of distant metastasis. There is still not a method of predicting and early diagnosing the metastasis. This study was conducted to investigate the relationship between the expression of multiple tumor suppressor 1(MTS1) as well as E-cadherin and metastasis in breast cancer. METHODS: The expression of MTS1 and E-cadherin were detected by SP immunohistochemical technique in 54 paraffin-embedded tissue specimens of breast cancer. RESULTS: The positive rates of MTS1 in the distant metastasis group and more than 4 local lymph node metastasis group were 40.7% and 38.1%, respectively, which were lower than individual corresponding control groups (74.1% and 69.7%) (P< 0.05). The MTS1 expression rates in the three groups of pathology grade I, II, and III were 72.6%, 58.8%, and 31.3%, respectively, with significant differences (P< 0.05). The positive rates of E-cadherin in the distant metastasis group and more than 4 local lymph node metastasis group were 37% and 23.8%, respectively, which are lower than individual corresponding control groups (70.4% and 66.7%) (P< 0.05). The E-cadherin expression rates in the three groups of pathology grade I, II, III were 80.9%, 47.1%,and 25%, respectively, with significant differences (P< 0.01). CONCLUSION: There was a close relation between MTS1, E-cadherin and invasion, metastasis of breast cancer. They can be used as the biological markers for evaluating the latent metastasis, disease development, and prognosis in breast cancer. PMID- 12753719 TI - [Expression of matrix metalloproteinase 9 (MMP-9) and laminin-receptor in breast carcinoma and their correlation with tumor metastasis and prognosis]. AB - BACKGROUND & OBJECTIVE: Previous researches of breast cancer mainly focused on its parenchyma and prognosis; however, the relationship between extracellular matrix of breast carcinoma and metastasis as well as prognosis was still unknown. The present study was designed to investigate the expression of matrix metalloproteinase 9 (MMP-9) and laminin- receptor (LM-R) in breast carcinoma and their correlation with tumor metastasis and prognosis. METHODS: The immunohistochemical method was applied to examine the expression of MMP-9 and LM R in 80 samples of breast carcinoma and 10 breast tissues of hyperplasia. The absorbance(A) values that indicate the expression levels were analyzed by computer image technique. t-test, chi square-test and other related tests were used to analyze their relationship with tumor metastasis and prognosis. RESULTS: The expression of MMP-9 in breast carcinoma was apparently higher than that in hyperplasia (t=8.87, P< 0.05). LM-R expressed at the basal membrane and the edge of some acini and ducts in hyperplasia tissues, while expressed at cytoplasm and cellular membrane in breast carcinoma tissues. LM-R expression of tumor in 17 cases with LM-R expression in basal membrane was higher than that in those without LM-R expression in basal membrane (t=2.02, P< 0.05). LM-R expression was related to histological grade; the lower the degree of differentiation, the higher the expression level (F=3.27, P< 0.05). The expression of MMP-9 and LM-R related to lymphatic metastasis. The expression levels of MMP-9 and LM-R in the group which had more than 3 involved lymph node were lower than those in the group which had less than involved 3 lymph node and no involved lymph node (t(MMP 9)=3.42, t(LM-R)=4.31, P< 0.05). The expression of MMP-9 and LM-R in original focus had significantly positive relation to metastasis(r(MMP-9)=0.654, r(LM R)=0.755, P< 0.001). MMP-9 had negative relation to progestogen receptor (PR) (r= 0.363,P< 0.05) and positive relation to LM-R (r=0.503,P< 0.01). The expression levels of MMP-9 and LM-R of the group which survived more than 3 years were lower than that in the group which survived less than 3 years (t(MMP-9)=3.57, t(LM R)=4.22, P< 0.05). Both of MMP-9 and LM-R were negatively related to the survival of the patients with breast carcinoma (r(MMP-9)=-0.387, r(LM-R)=-0.369, P< 0.05). CONCLUSION: MMP-9 and LM-R are useful indexes for predicting the metastasis and prognosis of breast cancer. PMID- 12753720 TI - [Role of tumor metastasis suppressor gene KAI1 in development of colorectal cancer]. AB - BACKGROUND & OBJECTIVE: The study of tumor metastasis suppressor gene KAI1 in colorectal cancer (CRC) is limited nowadays. There are some controversies about the expression of KAI1 and its relationship with pathologic classification of CRC, and up today, the reports of the correlation between KAI1 and the prognosis of CRC still be few. This study was designed to investigate the role of KAI1 in development of CRC and its value in predicting the prognosis of CRC. METHODS: The expression of KAI1 in 91 cases of primary CRC and 25 cases of lymph node metastases were determined using immunohistochemistry. RESULTS: The positive cases of KAI1 in primary CRC were 54 (59.3%), the weak positive cases were 22 (24.2%), the negative cases were 15 (16.5%). KAI1 expression reduced significantly in the cases with low differentiation, lymph node metastasis and distant metastatic tumor (P< 0.01). The positive expression rate of KAI1 in the patients who survived more than 5 years (67.14%) was higher than that in the patients who survived less than 5 years (33.33%) (P< 0.05). The 5-year survival rates of the patients with positive, weak positive, and negative expression of KAI1 decreased in turn (87.04%, 63.34%, 60.00%; P< 0.05). KAI1 expression in lymph node metastases was lower than that in primary locus (P< 0.05). CONCLUSION: The abnormal expression of KAI1 participates in malignant progression of CRC. Detecting the expression of KAI1 probably possesses clinical significance in evaluating the differentiation, lymph node metastasis,and distant metastasis of CRC, and predicting the prognosis of CRC. PMID- 12753721 TI - [Significance of urinary nucleosides in diagnosis of gastric carcinoma]. AB - BACKGROUND & OBJECTIVE: It was reported that urinary modified nucleosides with abnormally high amounts were found in many cancer patients. This study was designed to investigate the usefulness of urinary nucleosides in the diagnosis of gastric carcinoma. METHODS: The concentrations of 15 kinds of urinary nucleosides from 50 healthy persons and 48 patients with gastric carcinoma were determined by high-performance liquid chromatography (HPLC). Of 48 patients with gastric carcinoma, 25 underwent serum CEA examination. RESULTS: The average levels of 14 kinds of urinary nucleosides, m5U excepted, from patients with gastric carcinoma were higher than those from health persons (P< 0.05). Pseu 22.91+/-4.90, 34.87+/ 21.41; U 0.34+/-0.32, 0.62+/-0.82; A 0.58+/-0.16, 0.96+/-0.75; C 0.17+/ 0.15,0.24+/-0.19; m5U 0.03+/-0.07,0.07+/-0.06; I 0.26+/-0.10, 0.43+/-0.36; m1I 1.34+/-0.34, 2.44+/-1.39; ac4C 0.75+/-0.24, 1.08+/-0.72; G 0.09+/-0.04, 0.14+/ 0.10; X 1.20+/-0.42, 1.90+/-1.09; m2G 0.61+/-0.16, 1.00+/-0.69; m6A 0.04+/-1.13, 0.07+/-0.08; m1A 2.26+/-0.56, 3.71+/-2.21; m22G 1.34+/-0.27, 2.25+/-1.39; m1G 0.80+/-0.25, 1.41+/-0.86. The level of nucleoside I was positively correlated with the tumor size (P< 0.05). The level of nucleoside X was positively correlated with lymph node metastasis (P< 0.05). Using the concentrations of 15 nucleosides as the data vectors, principal component analysis was applied to classify gastric cancer patients and normal adults, 63%(30/48) of cancer patients were correctly classified, in which the identification rate was higher than that of CEA method (12%). CONCLUSION: Urinary modified nucleoside increased in the patients with gastric carcinoma, and it may be helpful in the diagnosis of gastric carcinoma. PMID- 12753722 TI - [Characteristics of occult cervical lymph node metastasis in squamous cell carcinoma of tongue and their influence on prognosis]. AB - BACKGROUND & OBJECTIVE: Occult cervical lymph node metastasis in squamous cell carcinoma of tongue has regularity and influence on the prognosis of patients. This study was designed to investigate the characteristics of occult cervical lymph node metastasis in squamous cell carcinoma of tongue and their influence on the prognosis to provide clinical bases for elective supraomohyoid neck dissection. METHODS: The data of 164 patients with squamous cell carcinoma of tongue who were treated in cancer center, Sun Yat-sen University, from 1990 to 1996 were reviewed. The characteristics of occult cervical lymph node metastasis in squamous cell carcinoma of tongue and their influence on the prognosis were analyzed. RESULTS: The rate of occult cervical lymph node metastasis of squamous cell carcinoma of tongue was 25.71%. The most common site of occult cervical lymph node metastasis was ipsilateral level II, and followed by ipsilateral level I and III. 82.98% of the lymph nodes of occult metastasis occurred in the above three levels. Most cervical lymph nodes of occult metastasis were found within two years after the first operation (33/36). There was significant difference of the prognosis between the group of dominant or occult cervical lymph node metastasis and the group of without cervical lymph node metastasis in squamous cell carcinoma of tongue (log-rank,P< 0.01); whereas there was no significant difference of the prognosis between the group of dominant cervical lymph node metastasis and the group of occult cervical lymph node metastasis according to log-rank test (P >0.05). CONCLUSION: The ipsilateral level I to level III were the common regions where occult cervical lymph node metastasis occurred. The authors suggest that elective supraomohyoid neck dissection may be applied to clinically negative neck of the patients with squamous cell carcinoma of tongue who were prone to metastasize latently. Occult cervical lymph node metastasis influences the prognosis of patients with squamous cell carcinoma of tongue significantly. PMID- 12753723 TI - [Research advances on p53 gene network]. AB - The p53 tumor suppressor gene integrates numerous signals to control cell life and death. p53 and its function-related genes consists of a complicated gene network. When a highly connected node in the network breaks down, the disruption of p53 has severe consequences. p53 gene locates in human chromosome 17q13.1. Its encoding wild-type p53 protein is composed of four parts: N-terminal activation domain, DNA-binding domain, oligomerization domain, and C-terminal regulation domain. As the gene structures of p73, p51, p63 are similar with p53, they are regarded as members of p53 gene family. p53 play an intermediate role connecting varied stress signals with the reactions of cells. DNA damage caused by ionizing radiation, aberrant growth signals, or chemotherapeutic drugs may activate the p53 network. When expression of p53 elevated, p53-mdm2 and p14(ARF)-mdm2 feedback loops can accurately regulate the expression level of p53, and the cooperation of p33ING1b gene is also needed in the process of p53 exerting normal function. Phosphorylation and acelylation are two important mechanisms to modulate p53 activity in vivo. Several dozen downstream genes are controlled directly by p53, and the activity of p53 falls into four categories: cell cycle inhibition, apoptosis, genetic stability, and inhibition of blood vessel formation. Elucidation of the function of p53 gene network will help to clarify the interaction mechanisms of p53 gene and its function-related genes. PMID- 12753724 TI - [Recent advances on molecular pathology of prostate carcinoma]. AB - The pathologic grade and clinical stage have some restrictions for the evaluation of the prognosis of prostate carcinoma. Recently, the function of genes related to apoptosis and tumor suppressor genes on the development, progression,and prognostic value of prostate carcinoma was paid close attention due to further research on the molecular pathology of prostate cancer. Overexpression of Bcl-2 was found in high malignant patients of prostate carcinoma and related to androgen refraction and resistance against anticancer agents as well. The mutation of p53 was found in prostatic intraepithelial neoplasia(PIN) and prostate cancer. p53 can be used as an independent prognostic factor for prostate cancer. The deletion of PTEN and p27 is an important negative factor of prognosis. Overexpression of p21 and p16 which are inhibition protein of cell cycle have effects on the formation and differentiation of prostate cancer. Fas/FasL system plays an important role in apoptosis of prostatic epithelial cells and takes part in the carcinogenesis of prostate. BRCA1 and p73 also have effects on the genesis and development of prostate cancer. PMID- 12753725 TI - Experimental estimation of the role of P-Glycoprotein in the pharmacokinetic behaviour of telithromycin, a novel ketolide, in comparison with roxithromycin and other macrolides using the Caco-2 cell model. AB - PURPOSE: The aim of this study was to examine the transport mechanism of telithromycin in comparison with erythromycin, azithromycin, clarithromycin and roxithromycin. METHODS: These antibiotics were examined in Caco-2 cell monolayers in order to demonstrate the potential involvement of P-GP in the absorption process, using verapamil as a P-GP competitor. A model using concentration equilibrium conditions was developed to delineate passive and active permeability components of telithromycin and roxithromycin transport in order to predict absorption in humans. RESULTS: Comparison of telithromycin Papp(AB)/Papp(BA) ratios with those of the other antibiotics indicated that an efflux pump was involved which limited the transport of the macrolides to a greater extent than that of telithromycin. Modulation of Caco-2 transport of these antibiotics by verapamil and their reciprocal effect upon verapamil transport confirmed the involvement of P-GP and demonstrated that two substrates of P-GP may increase the transport of each other. Under concentration equilibrium conditions, both roxithromycin and telithromycin exhibited high mean Papp values for passive diffusion which extrapolated to 88% and 77% predicted human absorption respectively, if the involvement of P-GP was ignored. Both Km and Vm values suggested that saturation of P-GP by telithromycin may occur at a lower dose level in humans than with roxithromycin (Km= 9.8 microM, Vm= 0.3 microM and Km= 45 microM, Vm= 1.1 microM, respectively). At 4.10(-5) M of either telithromycin or roxithromycin the passive flux was respectively 48% and 16% greater than the active efflux. CONCLUSIONS: The high absorption potential of telithromycin combined with the low Km and Vm values and the high dose level suggest that in humans the efflux pump may not limit ketolide absorption and that the interaction with other P-GP substrates may not significantly increase its oral absorption. PMID- 12753726 TI - Drug release characteristics of lipid based benzoporphyrin derivative. AB - PURPOSE: The purpose of this study was to examine the transfer of verteporfin (BPDMA) from its lipid based formulation to serum proteins. METHODS: As a result of BPDMA being confined to the lipid phase, it was found that fluorescence from the photosensitizer was highly concentration quenched. This phenomenon was used to demonstrate rapid transfer of lipid-based drug to various plasma components such as albumin and lipoproteins. Gel electrophoresis was used to show transfer of drug to lipoproteins. RESULTS: Loss of fluorescence quenching showed rapid transfer of the drug from its lipid based formulation to serum proteins. Gel electrophoresis showed that both the drug and phospholipid components were transferred to the lipoprotein fraction concurrently. The electrophoretic mobility of plasma lipoproteins was increased as a result of their interaction with lipid-based BPDMA. It was also shown that the lipid-based structures were readily destabilized in the presence of relatively low concentrations of plasma, and that liposomes of this lipid composition were highly unlikely to be found intact in the circulation following intravenous injection. CONCLUSIONS: Verteporfin is rapidly transferred from its lipid based formulation to serum proteins. This rapid transfer, particularly to lipoproteins, provides a mechanism for its rapid delivery to cells. PMID- 12753727 TI - A preliminary investigation of chitosan film as dressing for punch biopsy wounds in rats. AB - PURPOSE: To investigate the wound healing efficacy of two chitosan films, Chit-AA and Chit-LA, in comparison with a commercial preparation, Omiderm, using punch biopsy wounds in rats. METHODS: The punch biopsy wounds were created in the abdominal region of male Wistar rats. The films were evaluated in terms of transparency, flexibility, adherence property, ease of removal from wounds without damaging underlying tissues and fluid accumulation. In addition, the wounds were examined for dryness, exudation, contraction, period of epithelialization and scar formation. RESULTS: Chit-AA, Chit-LA and Omiderm films were comparable in terms of transparency, flexibility, adherence property, ease of film removal from wounds without damaging underlying tissues and fluid accumulation. Although there was no statistically significant difference in wound dryness and exudation between the film treated wounds and untreated wounds (Control), a significant difference was obtained in complete wound closure (t(100%)), period of epithelialization and scar formation. CONCLUSIONS: Both Chit LA and Chit-AA were able to promote wound healing with minimal scar formation. PMID- 12753728 TI - Co-encapsulation of two plasmids in chitosan microspheres as a non-viral gene delivery vehicle. AB - PURPOSE: The aims of this study are to encapsulate two different plasmid DNAs (pGL2 and pMK3) in the same microsphere structure and to investigate in vivo transfection characteristics of chitosan microspheres. Furthermore, the effect of formulation factors, such as chitosan concentration and plasmid DNA amount on in vitro properties of microspheres were studied. METHODS: Double plasmid-loaded chitosan microspheres were prepared by complex coacervation. Release studies were done in phosphate buffered saline at 37 degrees C and released plasmid DNA was determined spectrophotometrically. Integrity of plasmid DNAs was checked by agarose gel electrophoresis. For in vivo transfection studies, microspheres were injected into the muscle of the mice and expression of proteins (beta galactosidase and luciferase) was measured. RESULTS: High encapsulation efficiency was obtained with chitosan microspheres (90%). The size of particles was about 1.15 - 1.28 m. No dependence was observed between the size and formulation variables (chitosan concentration and the amount of plasmid). After encapsulation process, integrity of two plasmids did not change. Plasmid DNAs were continuously released from chitosan microspheres. Chitosan concentrations and plasmid amounts affected in vitro release properties. After intramuscular injection of double plasmids loaded microspheres into muscle of the mice, co expression was obtained. High beta-galactosidase and luciferase productions were determined with these microspheres after a long post-transfection period (12 weeks). CONCLUSIONS: Our results showed that two plasmids could be encapsulated in chitosan microspheres without affecting their structural and functional integrity. Thus, sustained and high protein production was obtained with these microspheres. PMID- 12753729 TI - Pharmaceutical approaches to colon targeted drug delivery systems. AB - PURPOSE: Although oral delivery has become a widely accepted route of administration of therapeutic drugs, the gastrointestinal tract presents several formidable barriers to drug delivery. Colonic drug delivery has gained increased importance not just for the delivery of the drugs for the treatment of local diseases associated with the colon but also for its potential for the delivery of proteins and therapeutic peptides. To achieve successful colonic delivery, a drug needs to be protected from absorption and /or the environment of the upper gastrointestinal tract (GIT) and then be abruptly released into the proximal colon, which is considered the optimum site for colon-targeted delivery of drugs. Colon targeting is naturally of value for the topical treatment of diseases of colon such as Chron's diseases, ulcerative colitis, colorectal cancer and amebiasis. Peptides, proteins, oligonucleotides and vaccines pose potential candidature for colon targeted drug delivery. METHODS: The various strategies for targeting orally administered drugs to the colon include covalent linkage of a drug with a carrier, coating with pH-sensitive polymers, formulation of timed released systems, exploitation of carriers that are degraded specifically by colonic bacteria, bioadhesive systems and osmotic controlled drug delivery systems. Various prodrugs (sulfasalazine, ipsalazine, balsalazine and olsalazine) have been developed that are aimed to deliver 5-amino salicylic acid (5-ASA) for localized chemotherapy of inflammatory bowl disease (IBD). Microbially degradable polymers especially azo crosslinked polymers have been investigated for use in targeting of drugs to colon. Certain plant polysaccharides such as amylose, inulin, pectin and guar gum remains unaffected in the presence of gastrointestinal enzymes and pave the way for the formulation of colon targeted drug delivery systems. The concept of using pH as a rigger to release a drug in the colon is based on the pH conditions that vary continuously down the gastrointestinal tract. Times dependent drug delivery systems have been developed that are based on the principle to prevent release of drug until 3-4 h after leaving the stomach. Redox sensitive polymers and bioadhesive systems have also been exploited to deliver the drugs into the colon. RESULTS: The approach that is based on the formation of prodrug involves covalent linkage between drug and carrier. The type of linkage that is formed between drug and carrier would decide the triggering mechanism for the release of drug in colon. The presence of azo reductase enzymes play pivotal role in the release of drug from azo bond prodrugs while glycosidase activity of the colonic microflora is responsible for liberation of drugs from glycosidic prodrugs. Release of drugs from azo polymer coated dosage forms is supposed to take place after reduction and thus cleavage of the azo bonds by the azoreductase enzymes present in the colonic microflora. Natural polysaccharides have been used as tools to deliver the drugs specifically to the colon. These polysaccharides remain intact in the physiological environment of stomach and small intestine but once the dosage form enters into colon, it is acted upon by polysaccharidases, which degrades the polysaccharide and releases the drug into the vicinity of bioenvironment of colon. However, they should be protected while gaining entry into stomach and small intestine due to enormous swelling and hydrophilic properties of polysaccharides. This has been achieved either by chemical crosslinking or by addition of a protective coat. Formulation coated with enteric polymers releases drug when pH move towards alkaline range while as the multicoated formulation passes the stomach, the drug is released after a lag time of 3-5 h that is equivalent to small intestinal transit time. Drug coated with a bioadhesive polymer that selectively provides adhesion to the colonic mucosa may release drug in the colon. CONCLUSIONS: Improved drug delivery systems are required for drugs currently in use to treat localized diseases of the colon. The advantages of targeting drugs specifically to the diseased colon are reduced incidence of systemic side effects, lower dose of drug, supply of the drug to the biophase only when it is required and maintenance of the drug in its intact form as close as possible to the target site. PMID- 12753730 TI - Development of liposomal polyene antibiotics: an historical perspective. AB - PURPOSE: The purpose of this review article is to review the development of a number of liposomal polyene antibiotics. BACKGROUND: In the past thirty years, the increase in life-threatening pre-systemic and systemic fungal infections within cancer, diabetic and AIDS patients have reached alarming proportions. A number of antifungal agents have been developed to combat this problem. In particular, polyene antibiotics such as Amphotericin B (AmB) and Nystatin (Nys) have remained the most effective and widely used agents in the treatment of these infections. However, their administration is limited by dose-dependent toxicities. One such dose-limiting toxicity is renal toxicity. Polyene antibiotic induced renal toxicity is believed to be mediated by the drug anchoring to cholesterol within the mammalian cell membrane, resulting in pore formation, abnormal electrolyte flux, decrease in adenosine triphosphate (ATP), and eventually a loss of cell viability. CONCLUSION: In the 1980s and 90s a number of promising lipid-based AmB and Nys formulations were developed to overcome these toxicities. This article will review the development of these liposomal polyene antibiotics. PMID- 12753731 TI - Cost-savings from subsidized pro-active pharmacist interventions. AB - PURPOSE: This paper evaluates a pilot project to determine the desirability of implementing a reimbursement model for pro-active interventions. A drug plan administration conducted an experiment in which a pharmacist could recommend to physicians the substitution of lower-cost therapies with equivalent health outcomes. The pharmacist shared any cost savings with the insurer. METHODS: Drug plan costs without the intervention were estimated using time-series forecasting models and compared to actual costs with the intervention. RESULTS: Over the course of this experiment, there were some cost savings generated by reactive pharmacist interventions but pro-active interventions, intended to influence subsequent physician behaviour, appear to have had no significant effect on the profile of drug expenditures. CONCLUSIONS: The evidence does not lend extensive support for full implementation of this type of reimbursement model. PMID- 12753733 TI - Hide and seek: the immunology of HSV persistence. AB - Herpes simplex virus (HSV) has been thought to persist as a latent infection by completely extinguishing antigen expression. In this issue of Immunity, Khanna et al. challenge this view by showing that CTL specific for antigens normally associated with active viral replication are crucial participants in regulating virus latency. PMID- 12753732 TI - Farnesol for aerosol inhalation: nebulization and activity against human lung cancer cells. AB - PURPOSE: A nebulized aerosol formulation of the anti-cancer agent farnesol is developed and shown to induce cell death of human lung cancer cells in vitro. METHODS: A nebulized farnesol formulation containing polysorbate 80 (Tween 80) is developed. The measurements of the aerosol properties during nebulization were used as input for a mathematical model of airway surface liquid in the lung of an average adult, to estimate the airway surface liquid drug concentration of the deposited farnesol. Cytotoxicity of the formulations was measured in vitro on non small cell lung cancer cells (H460 and A549). RESULTS: As much as 100% of lung cancer cytotoxicity can be achieved by using Pari LC Star and LC Plus nebulizers. The estimated airway surface liquid concentrations of the deposited farnesol reveal that the IC50 of the nebulized farnesol can be achieved over the entire tracheobronchial region, using the above Pari nebulizers with a volume fill of 5 ml. CONCLUSIONS: Drug concentrations higher than IC50 in the airway surface liquid are predicted with our methods, suggesting in vivo trials of a formulation may be warranted with these particular nebulizers. PMID- 12753734 TI - Contrasting urban and rural lifestyles of memory CD8+ T cells. AB - Memory CD8(+) T cells in lymphoid tissue exhibit an unexpectedly high apoptosis rate, while memory cells located in several nonlymphoid tissues do not. This may be because the lymphoid CD8(+) memory T cell repertoire is being continuously updated, while the tissue repertoire is more stable. PMID- 12753735 TI - From the yolk sac to the spleen: New roles for Notch in regulating hematopoiesis. AB - Although Notch receptors are widely expressed during hematopoiesis, their roles outside of the T cell lineage are not well characterized. Two reports in this issue of Immunity show that Notch1 and Notch2, respectively, are required to generate the earliest embryonic hematopoietic stem cells and splenic marginal zone B cells. Thus, different Notch receptors have specific and nonoverlapping functions that influence multiple hematopoietic lineages at various stages of development. PMID- 12753738 TI - Characterization of dendritic cells that induce tolerance and T regulatory 1 cell differentiation in vivo. AB - Active suppression is mediated by a subpopulation of CD4(+) T cells that prevents autoimmunity. However, the mechanisms involved in their differentiation in vivo are currently under intensive research. Here we show that in vitro culture of bone marrow cells in the presence of IL-10 induces the differentiation of a distinct subset of dendritic cells with a specific expression of CD45RB. These CD11c(low)CD45RB(high) DCs are present in the spleen and lymph nodes of normal mice and are significantly enriched in the spleen of IL-10 Tg mice. These natural or in vitro-derived DCs display plasmacytoid morphology and an immature-like phenotype, and secrete high levels of IL-10 after activation. OVA peptide-pulsed CD11c(low)CD45RB(high) DCs specifically induce tolerance through the differentiation of Tr1 cells in vitro and in vivo. Our findings identify a natural DC subset that induces the differentiation of Tr1 cells and suggest their therapeutic use. PMID- 12753737 TI - Herpes simplex virus-specific memory CD8+ T cells are selectively activated and retained in latently infected sensory ganglia. AB - This study challenges the concept that herpes simplex virus type 1 (HSV-1) latency represents a silent infection that is ignored by the host immune system, and suggests antigen-directed retention of memory CD8(+) T cells. CD8(+) T cells specific for the immunodominant gB(498-505) HSV-1 epitope are selectively retained in the ophthalmic branch of the latently infected trigeminal ganglion, where they acquire and maintain an activation phenotype and the capacity to produce IFN-gamma. Some CD8(+) T cells showed TCR polarization to junctions with neurons. A gB(498-505) peptide-specific CD8(+) T cell clone can block HSV-1 reactivation from latency in ex vivo trigeminal ganglion cultures. We conclude that CD8(+) T cells provide active surveillance of HSV-1 gene expression in latently infected sensory neurons. PMID- 12753739 TI - Impaired innate host defense causes susceptibility to respiratory virus infections in cystic fibrosis. AB - Viral infection is the primary cause of respiratory morbidity in cystic fibrosis (CF) infants. Here, we identify that host factors allow increased virus replication and cytokine production, providing a mechanism for understanding the severity of virus disease in CF. Increased virus is due to lack of nitric oxide synthase 2 (NOS2) and 2', 5' oligoadenylate synthetase (OAS) 1 induction in response to virus or IFNgamma. This can be attributed to impairment of activation of signal transducer and activator of transcription (STAT)1, a fundamental component to antiviral defense. NO donor or NOS2 overexpression provides protection from virus infection in CF, suggesting that NO is sufficient for antiviral host defense in the human airway and is one strategy for antiviral therapy in CF children. PMID- 12753740 TI - Virus-specific CD8 T cells in peripheral tissues are more resistant to apoptosis than those in lymphoid organs. AB - CD8 T cells persist at high frequencies in peripheral organs after resolution of an immune response, and their presence in the periphery is important for resistance to secondary challenge. We show here that LCMV-specific T cells in peripheral tissue (peritoneal cavity, lung, fat pads) reacted much less with the apoptotic marker Annexin-V than those in spleen and lymph nodes. This was not due to a TCR-based selection. In comparison to lymphoid tissue, T cells in the periphery expressed lower levels of Fas and Fas ligand and were resistant to activation-induced cell death in vitro. This may contribute to the survival of nondividing peripheral memory T cells, enabling them to efficiently function without being driven into apoptosis. PMID- 12753741 TI - CD4(+)CD3(-) accessory cells costimulate primed CD4 T cells through OX40 and CD30 at sites where T cells collaborate with B cells. AB - In this report we identify an accessory cell that interacts with primed and memory T cells at sites where they collaborate with B cells. These cells are distinguished from conventional dendritic cells by their lack of response to Flt3 ligand and their inability to process antigen. Unlike dendritic cells, the CD4(+)CD3(-) cells have little CD80 or CD86 expression but do express high levels of the TNF ligands, OX40 ligand and CD30 ligand. We show that Th2-primed cells express the receptors for these TNF ligands and preferentially survive when cocultured with these cells. Furthermore, we show that the preferential survival of OX40(+) T cells and support of memory T cell help for B cells are linked to their association with CD4(+)CD3(-) cells in vivo. PMID- 12753742 TI - Recruitment of TNF receptor 1 to lipid rafts is essential for TNFalpha-mediated NF-kappaB activation. AB - Engagement of TNF receptor 1 by TNFalpha activates the transcription factor NF kappaB but can also induce apoptosis. Here we show that upon TNFalpha binding, TNFR1 translocates to cholesterol- and sphingolipid-enriched membrane microdomains, termed lipid rafts, where it associates with the Ser/Thr kinase RIP and the adaptor proteins TRADD and TRAF2, forming a signaling complex. In lipid rafts, TNFR1 and RIP are ubiquitylated. Furthermore, we provide evidence that translocation to lipid rafts precedes ubiquitylation, which leads to the degradation via the proteasome pathway. Interfering with lipid raft organization not only abolishes ubiquitylation but switches TNFalpha signaling from NF-kappaB activation to apoptosis. We suggest that lipid rafts are crucial for the outcome of TNFalpha-activated signaling pathways. PMID- 12753743 TI - Competing functions encoded in the allergy-associated F(c)epsilonRIbeta gene. AB - Allergic reactions are triggered via crosslinking of the high-affinity receptor for immunoglobulin E, F(c)epsilonRI. In humans, F(c)epsilonRI is expressed as a tetramer (alphabetagamma(2)) and a trimer (alphagamma(2)). The beta subunit is an amplifier of F(c)epsilonRI surface expression and signaling. Here, we show that as a consequence of alternative splicing, the F(c)epsilonRIbeta gene encodes two proteins with opposing and competing functions. One isoform is the full-length classical beta, the other a novel truncated form, beta(T). In contrast to beta, beta(T) prevents F(c)epsilonRI surface expression by inhibiting alpha chain maturation. Moreover, beta(T) competes with beta to control F(c)epsilonRI surface expression in vitro. We propose that the relative abundance of the products of the beta gene may control the level of F(c)epsilonRI surface expression and thereby influence susceptibility to allergic diseases. PMID- 12753744 TI - Notch2 is preferentially expressed in mature B cells and indispensable for marginal zone B lineage development. AB - The Notch genes play a key role in cellular differentiation. The significance of Notch1 during thymocyte development is well characterized, but the function of Notch2 is poorly understood. Here we demonstrate that Notch2 but no other Notch family member is preferentially expressed in mature B cells and that conditionally targeted deletion of Notch2 results in the defect of marginal zone B (MZB) cells and their presumed precursors, CD1d(hi) fraction of type 2 transitional B cells. Among Notch target genes, the expression level of Deltex1 is prominent in MZB cells and strictly dependent on that of Notch2, suggesting that Deltex1 may play a role in MZB cell differentiation. PMID- 12753745 TI - HDAC7, a thymus-specific class II histone deacetylase, regulates Nur77 transcription and TCR-mediated apoptosis. AB - We report that HDAC7, a class II histone deacetylase, is highly expressed in CD4(+)CD8(+) double-positive thymocytes. HDAC7 inhibits the expression of Nur77, an orphan receptor involved in apoptosis and negative selection, via the transcription factor MEF2D. HDAC7 is exported from the nucleus during T cell receptor activation, leading to Nur77 expression. A triple HDAC7 mutant (S155A, S318A, S448A) is not exported from the nucleus in response to TCR activation and suppresses TCR-mediated apoptosis. Conversely, a fusion of HDAC7 to the transcriptional activator VP16 activates Nur77 expression. Inhibition of HDAC7 expression by RNA interference causes increased apoptosis in response to TCR activation. These observations define HDAC7 as a regulator of Nur77 and apoptosis in developing thymocytes. PMID- 12753746 TI - Notch1 but not Notch2 is essential for generating hematopoietic stem cells from endothelial cells. AB - Hematopoietic stem cells (HSCs) are thought to arise in the aorta-gonad mesonephros (AGM) region of embryo proper, although HSC activity can be detected in yolk sac (YS) and paraaortic splanchnopleura (P-Sp) when transplanted in newborn mice. We examined the role of Notch signaling in embryonic hematopoiesis. The activity of colony-forming cells in the YS from Notch1(-/-) embryos was comparable to that of wild-type embryos. However, in vitro and in vivo definitive hematopoietic activities from YS and P-Sp were severely impaired in Notch1(-/-) embryos. The population representing hemogenic endothelial cells, however, did not decrease. In contrast, Notch2(-/-) embryos showed no hematopoietic deficiency. These data indicate that Notch1, but not Notch2, is essential for generating hematopoietic stem cells from endothelial cells. PMID- 12753747 TI - Inducible transgenic mice reveal resting dendritic cells as potent inducers of CD8+ T cell tolerance. AB - Dendritic cells (DC) are inducers of immune responses par excellence. They also seem responsible for the induction of peripheral T cell tolerance. To investigate these opposite functions of DC, we generated a Cre/LoxP-based system that allows inducible antigen presentation by DC in vivo. This enables us to study the immunogical consequences of antigen presentation by resting versus mature DC without adoptively transferring DC and with physiological numbers of endogenous, naive responder T cells. We found that presentation of LCMV-derived CTL epitopes by resting DC resulted in antigen-specific tolerance, which could not be broken by subsequent infection with LCMV. On the other hand, antigen presentation by activated DC primed endogenous CTL to expand and to develop protective effector function. PMID- 12753748 TI - Terbutaline microparticles suitable for aerosol delivery produced by supercritical assisted atomization. AB - A new micronization technique called supercritical assisted atomization has been used to produce terbutaline microparticles with controlled particle size distribution in the range of drug particles deliverable by aerosol. The process is based on the solubilization of a fixed amount of supercritical carbon dioxide in a liquid solution; then, the ternary mixture is sprayed through a nozzle and atomized in order to produce microparticles. Water has been used as the liquid solvent; heated nitrogen has also been delivered into the precipitator to evaporate the liquid droplets. The process has been first optimized with respect to pressure and temperature (mixing temperature and pressure, precipitation temperature) and very mild operation conditions have been selected; then, the influence of the solute concentration in the liquid solution on particle size has been studied. The terbutaline produced powders were characterized with respect to morphologies and particle size. Spherical particles with very narrow volumetric particle size distributions were produced. Particularly, operating at 30 and 50mg of terbutaline per ml of water, more than 90% of the two distributions ranged between 1 and 3 microm; at 80 mg/ml more than 99% of the distribution ranged between 1 and 4 microm. HPLC analysis confirmed that no chemical degradation occurred in the drug as a consequence of the supercritical processing. PMID- 12753749 TI - Polyethylene glycol-coated liposomes for oral delivery of recombinant human epidermal growth factor. AB - The present study was to investigate the feasibility of oral delivery of recombinant human epidermal growth factor (rhEGF). Polyethylene glycol (PEG) coated liposomes containing rhEGF was prepared and evaluated for their stability and permeability in Caco-2 cells. In the animal study, we also determined plasma concentration and gastric ulcer healing effect after oral administration of rhEGF liposomes or the solution. Encapsulation of rhEGF into liposomes, suppressed the degradation in Caco-2 cell homogenate compared with the solution. The flux of rhEGF from dipalmitoylphosphatidylcholine (DPPC) liposome across Caco-2 cell monolayer from the apical to basolateral side was three times greater than that from phosphatidylcholine (PC) liposome or the solution. After oral administration of rhEGF liposomes or the solution in rats, the area under the concentration-time curve (AUC) of rhEGF increased 1.7- and 2.5-fold for PC and DPPC liposomes, respectively. The gastric ulcer healing effect was significantly increased in DPPC liposome compared with PC liposome and the solution. The enhanced curative ratio of rhEGF encapsulated into DPPC liposome may be due to the resistance to enzyme degradation, higher permeability and increased plasma AUC. Therefore, PEG coated liposomes containing rhEGF could be used as an oral delivery formulation with enhanced encapsulation efficiency. PMID- 12753750 TI - In vitro and in vivo evaluation of mucoadhesive microspheres consisting of dextran derivatives and cellulose acetate butyrate. AB - The objective of this study was to evaluate mucoadhesive properties and gastrointestinal transit of microspheres made of oppositely charged dextran derivatives and cellulose acetate butyrate (CAB). The microspheres were prepared by emulsion solvent evaporation method. A reference microsphere was made of lactose instead of dextran derivatives. Microspheres with a diameter of 425-710 microm were examined for in vitro mucoadhesion by the everted sac method. The results indicated that the percentage of adherence to the rat small intestine was affected by the amount of dextran derivatives in the microspheres. After 1.5h, the adhering percent of the reference microspheres and the microspheres containing 50% of dextran derivatives were 34 and 74%, respectively. Then gastrointestinal transit after oral administration to rats was evaluated by counting the microspheres remaining in the stomach and small intestine. The microspheres containing 40% of dextran derivatives adhered to the stomach rather than the small intestine. Mathematical analysis revealed that the time required for 50% of microspheres to leave the stomach was 1.42h, three times longer than the reference. These findings suggest that the microsphere is a promising device as a multiple-unit mucoadhesive system. PMID- 12753751 TI - Prediction of in vivo drug release behavior of controlled-release multiple-unit dosage forms in dogs using a flow-through type dissolution test method. AB - A newly designed flow-through type dissolution test method (FT method) was applied to predict in vivo drug release behaviors in dogs of controlled-release multiple unit dosage forms. The in vivo drug release behaviors were directly observed by measuring the residual amount of drugs in preparations recovered from the gastrointestinal (GI) tract after oral administration. Theophylline (TP), acetaminophen (AA), and phenylpropanolamine hydrochloride (PPA), which have different solubility, were used as model drugs. In vivo drug release behaviors in the gastrointestinal (GI) tract of dogs were similar to the results of the Wagner Nelson method. In vivo release behaviors of TP and AA, until 2h after administration, were well correlated to in vitro behaviors obtained by the paddle method at 100 rpm. However, the in vivo release rates of TP and AA were gradually decreased because of a lack of fluid in the lower region of the GI tract, their poor solubility, the difference of the release rates, and so on. Non-sink conditions, which would reflect TP and AA release in the lower region of the GI tract, were obtained by the FT method at a cell volume of 0.5 ml and a flow rate of 0.37 ml/h (TP), 0.48 ml/h (AA), respectively. The in vitro release profiles obtained by the FT method combining sink and non-sink conditions were similar to their in vivo profiles. On the other hand, in the case of PPA, the in vivo release profiles were considerably similar to the in vitro ones obtained by both the paddle method and the FT method. In conclusion, the FT method combining sink and non-sink conditions will give a good in vitro/in vivo correlation regarding release behavior for controlled-release multiple unit dosage forms. PMID- 12753752 TI - Stability and antitumor effects of all-trans retinoic acid-loaded liposomes contained sterylglucoside mixture. AB - The aim of this study was to develop an intravenous delivery of all-trans retinoic acid (ATRA) for the treatment of cancer. Two kinds of liposomes composed of dipalmitoylphosphatidylcholine and cholesterol with sterylglucoside (SG) mixture (SG liposomes) or without SG (non-SG liposomes) were prepared. A limited amount of ATRA was incorporated into liposomes approximately 3mol%. ATRA-loaded SG liposomes were more stable at 4 degrees C with light protection for 24 days than non-SG liposomes; maintaining 83.1% ATRA and the average diameter 198.5 nm (36 days), and in 80% rat serum for 120 min. SG seems to increase the ATRA-loaded efficiency in liposomes and stability of liposomes compared with cholesterol. The mean survival time of mice given SG liposomes (18.2 days) indicated a greater antitumor activity than saline (14.1 days) (P<0.001) without change of mean body weight. It is concluded that the current ATRA-loaded SG liposomes are potentially applicable for hepatic metastasis of M5076 tumor. PMID- 12753753 TI - Oral sustained delivery of paracetamol from in situ-gelling gellan and sodium alginate formulations. AB - The purpose of this study was to evaluate the potential for the oral sustained delivery of paracetamol of two formulations with in situ gelling properties. Oral administration of aqueous solutions of either gellan gum (1.0%, w/v) or sodium alginate (1.5%, w/v) containing calcium ions in complexed form resulted in the formation of gel depots in rabbit and rat stomachs as a consequence of the release of the calcium ions in the acidic environment. In vitro studies demonstrated diffusion-controlled release of paracetamol from the gels over a period of 6h. The bioavailability of paracetamol from the gels formed in situ in the stomachs of rabbits following oral administration of the liquid formulations was similar to that of a commercially available suspension containing an identical dose of paracetamol. PMID- 12753754 TI - In vitro characterization of jet-milled and in-situ-micronized fluticasone-17 propionate. AB - Particle properties are decisive for therapeutic efficiency of an inhaled pulmonary drug. Jet-milling as the common way for micronization of inhaled powder drugs shows several disadvantages such as a non-homogeneous particle size distribution and unnatural, thermodynamically-activated particle surfaces causing a high agglomeration behavior. For pulmonary use in a dry powder inhaler (DPI) beside a small particle size, a good de-agglomeration activity is required. In this study, fluticasone-17-propionate (FP) is in-situ prepared in a respirable particle size by a controlled crystallization technique. First, the drug is dissolved in acetone and precipitated by a solvent change method in the presence of a cellulose ether (HPMC) as stabilizing hydrocolloid. By rapidly pouring the drug solution into the polymer-rich water phase, the previously molecularly dispersed drug is associated to small particles and stabilized against crystal growth simultaneously by the presence of the hydrophilic polymer. This dispersion was then spray-dried. The mean particle size of the drug was around 2 microm and consequently in the respirable range. The physico-chemical properties of the in situ-micronized drug were compared to those of an unmilled and a jet-milled quality. Differences in the X-ray patterns and amorphous parts could be detected for the jet-milled but not for the in-situ-micronized drug. In addition, the aerodynamic behavior of the engineered and the jet-milled FP was analyzed using the FlowCaps inhaler as delivery device and compared to the commercial product Flutide Diskus. The fine particle fraction (FPF) (<5 microm) was increased four fold from approximately 9% for the jet-milled drug to approximately 40% for the in-situ-micronized drug when the pure drug powder was dispersed with the FlowCaps device. PMID- 12753755 TI - The determination of equilibrium constants, DeltaG, DeltaH and DeltaS for vapour interaction with a pharmaceutical drug, using gravimetric vapour sorption. AB - The application of gravimetric vapour sorption (GVS) to the characterisation of pharmaceutical drugs is often restricted to the study of gross behaviour such as a measure of hygroscopicity. Although useful in early development of a drug substance, for example, in salt selection screening exercises, such types of analysis may not contribute to a fundamental understanding of the properties of the material. This paper reports a new methodology for GVS experimentation that will allow specific sorption parameters to be calculated; equilibrium constant (K), van't Hoff enthalpy change (DeltaH(v)), Gibbs free energy for sorption (DeltaG) and the entropy change for sorption (DeltaS). Unlike other reports of such type of analysis that require the application of a specific model, this method is model free. The analysis does require that over the narrow temperature range of the study DeltaH(v) is constant and there is no change in interaction mechanism. PMID- 12753756 TI - Using deepest regression method for optimization of fluidized bed granulation on semi-full scale. AB - This study applied the deepest regression method to estimate the granule size of unsuccessful fluidized bed granulation runs. This study uses data from a previous study [Int. J. Pharm. 220 (2001) 149] on optimization of fluidized granulation process, wherein 8 of the 30 runs did not succeeded due to overwetting of the powder bed. The "complete data" (the observed and the estimated granule size by the depth regression method) were used to develop two regression models for the granule size: an empirical model based on the process variables (inlet air temperature, inlet airflow rate, spray rate, and inlet air humidity) and a fundamental model based on the powder bed moisture content and the relative droplet size. The regression models based on the incomplete data from the previous study and the regression models of the "complete data" were comparable in the sense that the contour plots based on the respective models and the predicted granule size were comparable. PMID- 12753757 TI - Structural modeling of drug release from biodegradable porous matrices based on a combined diffusion/erosion process. AB - Biodegradable, porous microspheres exhibit a wide range of release profiles. We propose in this paper a unifying approach based on the dual action of diffusion and erosion to establish which mechanisms are responsible for the variety of release kinetics observed during in vitro experiments. Our modeling procedure leads to the partitioning of the matrix into multiple, identical elements, thus simplifying significantly the mathematical and numerical treatment of the problem. The model equations cannot be solved analytically, since the domain contains a moving interface, and must therefore be solved numerically, using specific methods designed for that purpose. Our model confirms the major role that the relative dominance between diffusion and erosion plays in the release kinetics. In particular, the velocity of erosion, the effective diffusion coefficient of the drug molecule in the wetted polymer, the average pore length, and the initial pore diameter are sensitive parameters, whereas the porosity and the effective diffusion coefficient of the drug in the solvent-filled pores is seen to have little influence, if any, on the release kinetics. The model is confirmed by using release data from biodegradable microspheres with different ratios of low and high molecular weight PLA. Excellent goodness of fit is achieved by varying two parameters for all types of experimental kinetics: from the typical square root of time profile to zero-order kinetics to concave release curves. We are also able to predict, by interpolation, release curves from microspheres made of intermediate, untested ratios of PLA by using a relation between two model parameters. PMID- 12753758 TI - Crystallisation of amorphous mannitol is retarded using boric acid. AB - An approach to inhibit the crystallisation of amorphous mannitol was investigated. Boric acid was selected as a model additive for a fundamental study of its ability to retard crystallisation and to facilitate characterisation of the properties of the amorphous solid. At concentrations above 5% (w/w) of boric acid, the DSC scans indicated that a totally amorphous solid could be prepared by cooling the melted pre-mixture under ambient conditions. An increase in the glass transition temperature (T(g)) was observed with a corresponding increase in boric acid content, and their relationship was well fitted by the Gordon-Taylor equation. This result suggested that mannitol and boric acid mixed homogeneously. The crystallisation profiles of the resultant amorphous compositions were best described by the Avrami-Erofeev equation (n=1/3), which indicated that random nucleation and three-dimensional crystal growth was the best-fitting mechanism of this crystallisation. The activation energy of crystallisation decreased with increasing boric acid content, indicating that the temperature dependency for crystallisation decreased with increasing boric acid content. Furthermore, the rate of crystallisation at 30 degrees C for mannitol alone was 7000 times higher than that of mannitol containing 7.5% (w/w) of boric acid. PMID- 12753759 TI - The improved compaction properties of mannitol after a moisture-induced polymorphic transition. AB - We have previously shown that by exposing one form of mannitol to high relative humidity, a moisture-induced polymorphic transition of mannitol with a concurrent change in particle morphology occurs [Int. J. Pharm. 247 (2002) 69]. In this paper, we propose that if these changes occur during a wet-granulation procedure, it may be possible to make bring about an in situ size-reduction of mannitol with compaction property enhancement. Powder X-ray diffraction and scanning electron microscopy confirmed that a polymorphic transition (the delta form forming the beta form) had occurred on wet-granulation, and that a concomitant morphology change resulted in an agglomerate consisting of filament-like fine primary crystals (delta-granule). The aim of present study was to evaluate the compression properties of this agglomerate. The compact compressed with delta granules possessed a tensile strength 1.5 times higher than other mannitol samples. Heckel analysis indicated that the mannitol compression process proceeded by deformation without fragmentation and was thus particle size dependent. The delta-granule showed enhanced plastic deformability, due to its unique particle structure. Because the intrinsic compression properties of the polymorphs were similar, the primary particle size and specific surface area of mannitol were indicated to be the major contributing factors for the improved compaction behaviour, rather than the polymorphic transition. When using the delta-granule as an excipient for a tablet formulation containing a high amount of phenylpropanolamine hydrochloride (PPA) as a poorly compactable model drug, excellent tablets could be prepared without capping, whereas conventional mannitol produced capped tablets. PMID- 12753760 TI - Permeation of cyproterone acetate through pig skin from different vehicles with phospholipids. AB - The permeation of cyproterone acetate (CPA) from Derma Membrane Structure (DMS) creams and liposomal formulations was investigated. Standard diffusion experiments with dermatomed porcine skin were performed. The cumulative CPA amount permeated of the DMS creams was between 2.9 and 6.8 microg/cm(2) within 48 h. By addition of a phospholipid concentrate, the CPA permeation could be 2.6 fold further increased compared to the control DMS. A working temperature of 60 degrees C resulted in a change of the preparation and a higher permeation which could be confirmed by additional differential scanning calorimetry studies. In case of the liposomal formulations, the CPA permeation was strongly dependent on the lipid content. The higher the lipid content, the higher was the CPA permeation. Extruding procedures for decreasing the particle size of the liposomes resulted in a two-fold increase in CPA permeation compared to the unextruded liposomes. It is possible to control the CPA permeation by combining various formulations containing different phospholipids with saturated and unsaturated fatty acids. PMID- 12753761 TI - Particle size of liposomes influences dermal delivery of substances into skin. AB - In the present study, the influence of vesicle size on the penetration of two fluorescently labeled substances into the human skin was investigated. For the measurements either a hydrophilic fluorescent compound [carboxyfluorescein (CF)] or a lipophilic one [1,1'-dioctadecyl-3,3,3',3'-tertramethylindocarbo-cyanine perchlorate (DiI)] were encapsulated into vesicles. Liposomal formulations were prepared by extruding the vesicles through polycarbonate membrane filters with pores of different sizes. In vitro penetration studies into human abdominal skin were performed by using the Franz diffusion cell and a standardized skin stripping technique in attempt to find an optimum size for topical drug delivery by liposomes. Confocal laser scanning microscopy (CLSM) was used to visualize the effect of penetration ability of liposomal DiI. The maximum DiI fluorescence in the skin was observed with smaller liposomes of 71 nm diameter. The liposomes with a size of 120 nm diameter showed statistically enhanced penetration of CF into the skin as compared to larger ones. The results indicated that the CF penetration was inversely related to the size of the liposomes, which was confirmed by the data of the confocal laser scanning microscopy studies. PMID- 12753762 TI - Compactibility of agglomerated mixtures of calcium carbonate and microcrystalline cellulose. AB - The tablet tensile strength (T) of agglomerated mixtures of microcrystalline cellulose-Avicel PH 102 (MC), calcium carbonate (CC) and polyvinylpyrrolidone (Povidone, PVP), lubricated with magnesium stearate (MS), and formed under a compaction pressure (P(c)) ranging up to 618MPa has been determined. The compactibility was defined through: ln(-ln(1-T/T(max)))=Slope x lnP(c)+Intercept. MC/CC mixtures added of an agglutinant, before and after lubrication, show an important positive effect on their tablet tensile strength compared to a lineal relationship. This positive effect becomes smaller with decreasing compaction pressures. By different mixing methods, the higher the mixing efficiency the higher the compactibility, following the order: spray-dried>wet massing>tumble mixing. The compactibility of MC/CC/PVP spray-dried mixtures with calcium carbonate content from 20 to 60% was equal to or greater than that of pure microcrystalline cellulose. After lubrication with 2% MS the compactibility decreased, only the mixture with the maximal tablet tensile strength attained the tensile strength of pure microcrystalline cellulose. The presence of the binder, the lubricant and higher compaction pressures allow the accommodation of higher calcium carbonate proportions in the mixtures, at the maximal tablet tensile strength of the series. The lubricant decreases in a greater extent the compactibility of mixtures with a continuous phase of MC/PVP than that of CC/PVP. This is attributed to the plastic behavior of the MC/PVP continuous phase compared to a calcium carbonate continuous phase able to disrupt the Povidone and the possible lubricant coatings allowing a stronger interparticle interaction. PMID- 12753763 TI - Poly(acrylic acid) microgels (carbopol 934)/surfactant interactions in aqueous media. Part I: nonionic surfactants. AB - The interaction between Tween 80 and Pluronic F-127 with carbopol in water was studied as a function of surfactant concentration. 0.25% carbopol microgels dispersions showed a continuous decrease in transmittance, viscosity and conductivity when surfactant concentration ranged from 0.01-0.02% to 0.50% Tween 80 or from 0.03-0.06% to 0.30% Pluronic F-127. These limit values can be considered as the critical association concentration and the saturation binding concentration, respectively. In this concentration range, a strong rise in pH (from 3.18 to 3.50) suggested that surfactant-polymer binding occurred mainly through a stoichiometric hydrogen-bonding interaction between the oxyethylene and carboxylic groups. In the presence of carbopol, the concentration of Tween 80 at the air/water interface decreases as the surfactant is adsorbed onto the polymer and drawn into the bulk solution. In contrast, the interaction with the polymer seems to change the conformation of the expanded chains of Pluronic F-127, making it easier for more molecules of surfactant to be at the interface and increasing the thickness of the interfacial surfactant layer. Fluorescence probes indicated that the carbopol network presents a more apolar medium than pure water, and the differences in the hydrophile-lipophile balance (HLB) of each surfactant were responsible for the lower I(I)/I(III) values obtained with Tween 80/carbopol systems. Microcalorimetry titration data made it possible to conclude that Tween 80/carbopol interaction, at 298K, is an enthalpy-driven process due to stabilization of Tween 80 units inside the polymer network. In contrast, Pluronic F-127/carbopol association (endothermic process) occurs owing to a gain in entropy when polymer-surfactant interaction allows the restoration of free water hydrogen-bonding structure, resembling the micellization process. PMID- 12753764 TI - Poly(acrylic acid) microgels (carbopol 934)/surfactant interactions in aqueous media. Part II: Ionic surfactants. AB - The interaction of sodium dodecylsulfate (SDS) or benzalkonium chloride (BkCl) with carbopol microgels (0.25% (w/w)) in water was studied through pH, trasmittance, viscosity, surface tension, conductivity, fluorescence, oil solubilization, and microcalorimetry measurements. In the case of the anionic surfactant, enthalpy-driven hydrophobic absorption of SDS into carbopol microgels began when SDS concentration reached 0.05-0.08% and ended around 0.6%. These concentrations were estimated as the critical aggregation (cac) and saturation binding concentration, respectively. The hydrophobic absorption of the surfactant accompanied by its counter-ion caused carbopol microgels to swell and promoted the occurrence of bridges among several carbopol microgels. As a consequence, the consistency of the dispersions increased significantly. Above binding saturation, further addition of SDS produced a shielding effect among the anionic charges of carbopol and its dehydration, which was shown as a decrease in the viscosity of the dispersions. At low shear rates, the dispersions behaved as pseudoplastic owing to orientation of carbopol/SDS aggregates in the flow direction. Increasing shear rates caused the inter-microgel bridges to break, the water layer surrounding them to diminish, and the system to show a shear-thickening behavior. In contrast, carbopol/BkCl aggregates showed shear-thickening flow in the whole range of shear rates analyzed. Electrostatic interactions between BkCl and carbopol carboxylic groups release protons to the medium and decrease the internal osmotic pressure of the microgels. This may favor the establishment of hydrophobic interactions among surfactant tails, and induces carbopol microgels to collapse. The cac was approximately 0.01% BkCl. Saturation binding occurred at 0.3-0.5%, indicating that only 25-40mol% carboxylic groups were neutralized with BkCl. The shrinking of carbopol microgels as BkCl is absorbed prevents additional surfactant molecules from interacting with the remaining carboxylic groups. Microcalorimetry assays revealed that the aggregation process occurred with a strong gain in enthalpy. PMID- 12753765 TI - Solubility of drugs in aqueous solutions. Part 1. Ideal mixed solvent approximation. AB - The present paper deals with the application of the fluctuation theory of solutions to the solubility of poorly soluble drugs in aqueous mixed solvents. The fluctuation theory of ternary solutions is first used to derive an expression for the activity coefficient of a solute at infinite dilution in an ideal mixed solvent and, further, to obtain an equation for the solubility of a poorly soluble solid in an ideal mixed solvent. Finally, this equation is adapted to the solubility of poorly soluble drugs in aqueous mixed solvents by treating the molar volume of the mixed solvent as nonideal and including one adjustable parameter in its expression. The obtained expression was applied to 32 experimental data sets and the results were compared with the three parameter equations available in the literature. PMID- 12753766 TI - Importance of using solid lipid microspheres as carriers for UV filters on the example octyl methoxy cinnamate. AB - The aim of this study was to prepare solid lipid microspheres (SLM) of octyl methoxy cinnamate (2-ethylhexyl-p-methoxy cinnamate; OMC) to achieve controlled release, decrease penetration of this UV absorber from skin and improve its photostability. The influence of the carrier on the rate of release was studied in vitro with a cellulose acetate membrane and in vivo from excised rat skin with Franz diffusion cells. The release rate was decreased by up to 13-80% with the SLM formulation. In vivo, penetration of OMC into skin was investigated by HPLC method. It was found out that the rate of penetration is significantly dependent upon the formulation and could be decreased by up to 77% in SLM formulations. When different topical vehicles were compared, OMC was released and penetrated into rat skin more quickly and in greater amount from vehicles containing free OMC than in SLM form. Additionally, photostability was shown to be improved in SLM form. PMID- 12753767 TI - Effect of chitosan on progesterone release from hydroxypropyl-beta-cyclodextrin complexes. AB - An inclusion complex composed of progesterone (Prog) and hydroxypropyl-beta cyclodextrin (HPbetaCD) was prepared by the spray-drying and freeze-drying methods. Prog alone and its inclusion complex with HPbetaCD were incorporated into chitosan by spray-drying and freeze-drying. The inclusion complex was characterized by IR and DSC. The inclusion complex was investigated in solution by phase solubility diagrams and stability constant was determined at pH 7.4 and at different temperatures (10, 25 and 37 degrees C) to obtain the thermodynamic parameters of inclusion. The results indicate that the Prog-HPbetaCD inclusion complex is more water soluble than Prog alone. Release data from all samples showed significant improvement of the dissolution rate of Prog and a controlled release is obtained in the presence of chitosan. PMID- 12753768 TI - Comparison of skin permeation enhancement by 3-l-menthoxypropane-1,2-diol and l menthol: the permeation of indomethacin and antipyrine through Yucatan micropig skin and changes in infrared spectra and X-ray diffraction patterns of stratum corneum. AB - 3-l-Menthoxypropane-1,2-diol (MPD) is a derivative of l-menthol, which has an enhancement effect on drug permeation through skin. In this study, the effect of MPD on drug permeation through skin was compared with that of l-menthol. MPD or l menthol at final concentrations of 3% in 40% ethanol was added to the drugs indomethacin or antipyrine and each mix then applied to Yucatan micropig skin in vitro. Drug concentrations in the skin were higher in the presence of either MPD or l-menthol, however, only l-menthol shortened the lag time of permeation. MPD enhanced the skin permeation of the drugs only by increasing the skin concentration of the drugs. In contrast, l-menthol enhanced the skin permeation of the drugs by increasing both the skin concentration and the diffusion rate in skin. The infrared (IR) spectra and X-ray diffraction patterns of stratum corneum after treatment with MPD did not differ from those of intact stratum corneum. A change in the IR spectra of stratum corneum after treatment with l-menthol was observed at the CH band, and the peaks representative of the lipid structure in the X-ray diffraction patterns decreased in intensity. These results suggest that l-menthol, but not MPD, disrupts the intercellular lipid structure of stratum corneum. Thus, MPD is expected to be a moderate skin permeation enhancer. PMID- 12753769 TI - Planar gamma scintigraphy--points to consider when quantifying pulmonary dry powder aerosol deposition. AB - Methodological aspects of planar gamma scintigraphy used to quantify pulmonary aerosol deposition were investigated using an experimental dry powder formulation. Particles of micronized salbutamol sulphate were labelled with technetium-99m and admixed to an ordered mixture of unlabelled micronized salbutamol sulphate and larger carrier particles of lactose. The radioaerosol was administered to 24 healthy subjects, 12 in each of two consecutive, similarly designed studies. Pulmonary deposition was determined using two methods: repeated planar imaging, and pharmacokinetic assessments following charcoal co administration to prevent gastrointestinal salbutamol absorption. After due consideration had been taken to ensure appropriate radiolabelling, image acquisition and processing procedures, a scintigraphic estimate of 26.2% (with 95% confidence interval of 24.2-28.4%) was obtained, which did not significantly differ from the pharmacokinetic estimate of 26.4% (24.4-28.7%). In summary, pre study validation of the radiolabelling technique, quality control of radioaerosols produced during the study, correction for re-distribution of radiolabel from the lungs, selection of regions of interest, assessment of lung contours, correction for tissue attenuation of gamma rays and establishment of the actual recovery of radioactivity in the scintigraphic measurements could potentially affect the accuracy of the scintigraphic estimate of pulmonary deposition and, thus, should be carefully considered in the design or evaluation of any such study. PMID- 12753770 TI - Enhancement of reactive oxygen species production from canine blood leukocytes by human recombinant interleukin-12. AB - A novel biological activity of human recombinant interleukin-12 (rhIL-12) on canine peripheral blood mononuclear cells (PBMC) was investigated in vitro. Canine PBMC were cultured in the presence or absence of rhIL-12 for 3 days. The reactive oxygen species (ROS) production induced by opsonized-zymosan (OZ) was then measured by a luminol-dependent chemiluminescense assay and demonstrated that the ROS production was enhanced after culture with rhIL-12. A nitro blue tetrazolium test and flowcytometry analysis revealed that canine lymphocytes, eosinophils, and monocytes were capable of ROS production, but that monocytes had the highest capacity. These results suggest that rhIL-12 enhances ROS production from canine monocytes. PMID- 12753771 TI - Cytokine profiles of peripheral blood mononuclear cells from dogs experimentally sensitized to Japanese cedar pollen. AB - Japanese cedar (Cryptomeria japonica, CJ) pollinosis is mediated by type-I hypersensitivity and induces seasonal rhinitis and conjunctivitis in humans. Previous studies showed that dogs could be experimentally sensitized with CJ pollen. In this study, we carried out quantitative analysis of mRNA levels of various cytokines in the peripheral blood mononuclear cells (PBMC) of 12 dogs experimentally sensitized to Japanese cedar pollen. Experimental sensitization was carried out by injection of crude CJ pollen extract with aluminium hydroxide gel. The expression levels of interleukin (IL)-1beta, IL-2, IL-4, IL-6, IL-8, IL 10, IL-18, interferon (IFN)-gamma, transforming growth factor (TGF)-beta(1), and tumor necrosis factor (TNF)-alpha mRNAs in the PBMC were quantified using a real time sequence detection system. In the PBMC tested without culture, the expression levels of IL-8 and TNF-alpha mRNAs in experimentally sensitized dogs were significantly higher than those in control dogs. The expression level of IFN gamma mRNA in the sensitized group was significantly lower than that in the control group. When the PBMCs were cultured in the presence of CJ pollen extract, the level of IL-4 mRNA expression was markedly increased in the PBMC from the experimentally sensitized dogs. In the PBMC stimulated with the CJ pollen extract, the expression level of IL-2 mRNA in the sensitized group was also significantly higher than that in the control group. Our data indicated that a Th2 response and proliferation of PBMC occur in response to the sensitizing antigen in dogs experimentally sensitized with CJ pollen, and revealed the presence of antigen-specific Th2 cells in this canine model. In addition, the expression levels of the mRNAs encoding proinflammatory cytokines were shown to be elevated after CJ pollen sensitization, indicating the activation of monocytes and macrophages. PMID- 12753772 TI - Alpha4-integrin (CD49d) expression on bovine peripheral blood neutrophils is related to inflammation of the respiratory system. AB - Neutrophil emigration from the pulmonary vasculature, is mediated by cellular adhesion molecules (CAM) expressed on the outer membranes of endothelial cells and neutrophils. Although beta(2)-integrin-dependent migration is a major mechanism of neutrophil migration, which was demonstrated by extensive invasion of neutrophils in pulmonary tissue of calves suffering from a genetic deficit in expression of beta(2)-integrins, termed bovine leukocyte adhesion deficiency (LAD), the role of alternative CAM is still unclear. We investigated whether an alternate CAM for beta(2)-integrin function, i.e. the alpha(4)-integrin, was expressed on peripheral blood neutrophils of calves. As we detected basal but significant expression, the effect of naturally acquired pulmonary infection on the expression of either integrin was determined, as an indication for its function in the migration process. In our experiments, basal expression of alpha(4)-integrins on peripheral blood neutrophils from clinically healthy calves was detected. On neutrophils of calves, experiencing field outbreaks of enzootic bronchopneumonia, higher expression of the alpha(4)-integrin was detected, which returned to normal after successful treatment of the disease. In addition, its level of expression was linearly related to plasma acute phase protein (haptoglobin) concentrations, which is a sensitive parameter for severity of respiratory inflammation. Increased expression of the alpha(4)-integrin on peripheral blood neutrophils during pulmonary inflammation indicates a role for this CAM in neutrophil migration in the lung. PMID- 12753773 TI - Development of a Staphylococcus aureus vaccine against mastitis in dairy cows. I. Challenge trials. AB - A vaccine composed of three field isolates of Staphylococcus aureus (S. aureus) derived from cases of mastitis in cows was developed. The vaccine was administered to nine uninfected cows while 10 other cows were used as controls. All cows were challenged with a highly virulent S. aureus strain administered into two quarters of each cow. Quarters were tested for clinical signs, secretion of S. aureus, and somatic cell count (SCC). No systemic effects were observed in any of the cows, vaccinated or control. Vaccinated cows had 70% protection from infection compared with fewer than 10% in the controls. Moreover, all quarters challenged in the vaccinated cows, regardless of whether they were successfully infected or not with S. aureus, exhibited very mild inflammatory reactions, identified by their low SCCs (<100,000). PMID- 12753774 TI - Characterization of efferent lymph cells and their function following immunization of cattle with an allogenic Theileria annulata infected cell line. AB - Immunization of cattle with in vitro propagated bovine mononuclear cells infected with Theileria annulata induces a protective immune response. Activation and effector function of T cells exiting the lymph node draining the site of cell line immunization were investigated to understand the mechanisms involved in the generation of immunity. Immunized animals exhibited a biphasic immune response in efferent lymph as well as peripheral blood. The first phase corresponded to allogenic responses against MHC antigens of the immunizing cell line and the second was associated with parasite specific responses. An increase in the output of CD2(+) cells and MHC class II(+) cells in efferent lymph was observed after cell line immunization with a corresponding decrease in WC1(+) cells. Although the percentage of CD4(+) T cells did not change significantly over the course of the experiment, they became activated. Both CD25 and MHC class II expressing CD4(+) T cells were detected from day 7 onwards, peaking around day 13. Efferent lymph leukocytes (ELL) exhibited sustained responses to IL-2 in vitro following cell line immunization. Antigen specific proliferation was also detected first to the immunizing cell line and then to parasite antigens. The two peaks of CD2(+) cells were observed, which corresponded to similar peaks of CD8(+) cells. The increase in CD8(+) cells was more pronounced during the second parasite specific phase than the first allogenic phase. Activated CD8(+) T cells mainly expressed MHC class II and some expressed CD25. Significantly the peak of activated CD4(+) T cells preceded the peak of activated CD8(+) T cells, highlighting the role of T. annulata specific CD4(+) T cells in inducing parasite specific CD8(+) cytotoxic responses. A biphasic cytotoxic response also appeared in efferent lymph and peripheral blood, the first directed against MHC antigens of the immunizing cell line followed by MHC class I restricted parasite specific cytotoxicity. The cytotoxic responses in efferent lymph appeared earlier than peripheral blood, suggesting that activated CD8(+) cells exiting the draining lymph node following immunization with T. annulata infected schizonts play an important role in the development of protective immune responses. PMID- 12753775 TI - Comparison of intradermal and serum testing for allergen-specific IgE using a Fcepsilon RIalpha-based assay in atopic dogs in the UK. AB - Atopic dermatitis in dogs is a common allergic skin disease that affects substantial numbers of dogs in the UK. The purpose of this study was to compare the results of an intradermal test (IDT) and an in vitro test in a large cohort of dogs. Dogs were intradermal tested with Greer allergens (Greer Labs Inc, Lenoir, NC, USA) using standard techniques. At the same time blood samples were drawn and submitted for evaluation by ELISA using the ALLERCEPT Definitive Allergen Panels for allergen-specific IgE, a commercial assay that uses a biotinylated recombinant extracellular domain of the high affinity Fc-epsilon receptor alpha chain protein (Fcepsilon RIalpha). The allergens used in the two tests included grass, tree and weed pollens, moulds, flea saliva/whole flea extract and house dust mite species. The optical density readings from the ELISA for each allergen were compared with the results of the IDT for 265 dogs. The prevalence of positive reactions in the ELISA was equal to or greater than the results of the IDT in the case of almost all of the allergens, but two notable exceptions were the house dust mites Dermatophagoides farinae and Dermatophagoides pteronyssinus. These two allergens were the most common positive reactions by IDT (prevalence D. farinae 78.9%, D. pteronyssinus 66.4%). The results of the two tests were significantly different (McNemar's test, P<0.05) for 16 of the 22 allergens. The sensitivities of the ELISA compared to the IDT (where there were more than 3 dogs with positive reactions in both tests) varied between 19.3 and 77.1% (D. pteronyssinus 19.3% and D. farinae 67.9%) and the specificities varied between 64.2 and 96.6% (D. pteronyssinus 96.6% and D. farinae 89.3%). PMID- 12753776 TI - Mycobacterium bovis AN5 antigens vary in their ability to induce nitric oxide production in blood monocytes of experimentally infected cattle. AB - Reactive nitrogen intermediates (RNI) are the principal effector molecules of activated monocyte/macrophage populations, responsible for killing and inhibiting the growth of virulent mycobacteria. In vitro nitrite production by blood monocytes of cattle inoculated with live Mycobacterium bovis AN5 was assessed from 0 day through 45 weeks post inoculation (PI). High in vitro nitrite production was observed at the 8th and 12th weeks PI in sensitized cattle but reactivity had fallen by the 20th week PI. To assess the in vitro nitrite producing ability of monocytes induced by individual polypeptides within culture filtrate antigens (CFA) of M. bovis AN5, cellular blotting was performed using peripheral blood mononuclear cells (PBMC) at the 12th week PI. It was observed that polypeptides of MW 70, 65, 60, 25, 24 and 22 kDa of CFA induced high nitrite production by blood monocytes while many polypeptides had little or no effect. PMID- 12753777 TI - Expression and functional characterization of killer whale (Orcinus orca) interleukin-6 (IL-6) and development of a competitive immunoassay. AB - Interleukin-6 (IL-6) is a cytokine that can reach detectable systemic levels and is a major inducer of the acute phase response. As such, clinical assays to identify this cytokine in mammalian sera are of diagnostic value. A 558 base-pair (bp) fragment of killer whale IL-6 was cloned and expressed as a 21 kDa protein in Escherichia coli. Biological activity of the recombinant killer whale IL-6 (rkwIL-6) was demonstrated using the IL-6-dependent B9 mouse hybridoma cell line; acute phase sera from a killer whale and supernatants from lipopolysaccharide (LPS)-stimulated killer whale peripheral blood mononuclear cells (PBMCs) also supported the proliferation of the B9 hybridoma. Rat anti-mouse IL-6 receptor antibody effectively blocked biological activity of all three sources of IL-6. Polyclonal antisera, specific for the recombinant protein, were obtained by successive immunization of a rabbit with rkwIL-6. The polyclonal antibody was capable of neutralizing the biological activity of both recombinant and native kwIL-6. A competitive enzyme-linked immunosorbent assay (ELISA) was developed using the polyclonal rabbit anti-rkwIL-6 and the recombinant protein; sensitivity of the assay was in the range of 1 ng/ml. The ELISA was subsequently used to identify the presence of native IL-6 in acute phase sera of two species of delphinidae, a killer whale and a bottlenose dolphin. The application of quantitative cytokine assays as diagnostic tools for monitoring cetacean health are becoming feasible as many animals are now being trained for fluke presentation, making blood collection a routine procedure. PMID- 12753778 TI - Comparison of hybrid and purebred in vitro-derived cattle embryos during in vitro culture. AB - Frozen-thawed spermatozoa collected from a beef bull (Japanese Black) were used for in vitro fertilization (IVF) of matured oocytes obtained from dairy (Holstein) and beef (Japanese Black) females. Embryos were examined for fertilization, cleavage rate, interval between insemination and blastocyst production (experiment I), total cell number per embryo and sex ratio during blastocyst formation (experiment II), and blastocyst production rate of zygotes that developed to 2-, 4-, and 8-cell stages at 48h post-fertilization (experiment III). Fertilized oocytes were cultured in vitro on a cumulus cell co-culture system. The fertilization and cleavage rate of oocytes groups were similar, however, the blastocyst production rate was greater (P<0.05) in hybrid than from purebred embryos (27% versus 20%). Development of blastocysts produced from hybrid embryos developed at a faster rate than blastocysts produced from the straightbred embryos. In hybrid embryos, blastocyst production was significantly greater on day 7 (56%) and gradually decreased from 20% on day 8 to 17% on day 9. In contrast, blastocyst production rate from the purebred embryos was lower on day 7 (17%), increasing on day 8 to 59% and then decreased on day 9 to 24%. The total number of cells per embryo and sex ratio of in vitro-produced blastocysts were not different between hybrid and purebred embryos. The number of blastocysts obtained from embryos at the 8-cell stage of development by 48h post fertilization (94%) was greater (P<0.01) than the number of zygotes producing blastocysts that had developed to the 4-cell stage (4%) and the 2-cell stage (2%) during the same interval. These results show that the blastocyst production rate and developmental rate to the blastocyst stage were different between hybrid and purebred embryos, and that almost all of the in vitro-produced blastocysts were obtained from zygotes that had developed to the 8-cell stage 48h post fertilization. PMID- 12753779 TI - Differences between Brahman and Holstein cows in response to estrus synchronization, superovulation and resistance of embryos to heat shock. AB - Embryos from Bos indicus are more resistant to elevated culture temperature (i.e. heat shock) than embryos from some Bos taurus breeds. The present experiment was designed to determine if Brahman embryos have greater resistance to heat shock than Holstein embryos at a stage in development before the embryonic genome was fully activated. A second objective was to test breed effects on estrus synchronization and superovulation responses. A total of 29 Brahman and 24 Holstein cows were subjected to estrus synchronization using gonadotropin releasing hormone (GnRH) and prostaglandin F2alpha (PGF2alpha) superovulation. Embryos were collected at 48 h and day 5 after insemination. There was a tendency for a lower proportion of Brahmans to be detected in standing estrus than Holsteins. There were no differences between breeds in the proportion of cows detected in estrus using both tailpaint and standing estrus as criteria or in interval from PGF2alpha to estrus. The degree of synchrony in estrus was greater for Brahmans. Superovulation response was generally similar between breeds. At 48 h after insemination, there was a tendency for a greater proportion of Brahman oocytes to have undergone cleavage. Uncleaved oocytes were cultured for an additional 24 h-at this time, cleavage rate was similar between breeds. When embryos reached the 2-4-cell stage, they were heat-shocked for 4.5 h at 41 degrees C. This heat shock reduced the proportion of embryos that developed to the blastocyst stage but there was no breedxtreatment interaction. At day 5 after insemination, the number of embryos recovered was too low to allow comparison of breed effects. In conclusion, genetic effects on cellular thermotolerance that make Brahman embryos more resistant to heat shock are not expressed at the 2-4 cell stage. There were few differences between Brahman and Holstein in response to estrus synchronization and superovulation. The fact that cleavage tended to occur earlier in Brahman than Holstein embryos suggests breed differences in timing of ovulation, fertilization or events leading to cleavage. PMID- 12753780 TI - Effect of neuropeptide Y on GnRH-induced LH release from bovine anterior pituitary cell cultures derived from heifers in a follicular, luteal or ovariectomized state. AB - Objectives were to determine if neuropeptide Y (NPY) had direct effects GnRH induced secretion of LH from the anterior pituitary gland, and if endogenous steroids modulated the effect of NPY. To accomplish these objectives, 15 Hereford heifers were assigned to one of three ovarian status groups: follicular, luteal, or ovariectomized. One animal from each of the three ovarian status groups was slaughtered on each of 5 days and anterior pituitary gland harvested. Anterior pituitary gland cells within ovarian status were equally distributed and randomly assigned to one of three cell culture treatments: no NPY or GnRH (control), 10 nM GnRH, or 100 nM NPY+10 nM GnRH. Anterior pituitary cell cultures were incubated with or without NPY for 4 h and further incubated for an additional 2 h with or without GnRH and supernatant collected for quantification of LH. Treatment of anterior pituitary cell cultures with GnRH or GnRH+NPY did not affect LH release in cultures obtained from follicular (S.E.=5%; P=0.58) or ovariectomized (S.E.=7%; P=0.22) heifers. Both GnRH and GnRH+NPY increased LH release from anterior pituitary cell cultures from heifers in the luteal phase (S.E.=14%; P < or = 0.05) compared to control cultures. Cultures from luteal phase heifers treated with GnRH did not differ from those treated with GnRH+NPY (P=0.34). These data provide evidence to suggest that effects of NPY on LH release may occur primarily at the level of the hypothalamus. PMID- 12753781 TI - Validation of a sensitive enzymeimmunoassay for 13,14-dihydro-15-keto-PGF2alpha in buffalo plasma and its application for reproductive health status monitoring. AB - A simple, sensitive and direct enzymeimmunoassay (EIA) procedure on microtitre plates using the second antibody coating technique was standardized and validated for the determination of 13,14-dihydro-15-keto PGF2alpha (PGFM) in unextracted buffalo plasma. The assay was carried out directly in 20 microl of buffalo plasma. PGFM standards prepared in charcoal stripped hormone-free plasma were used. The sensitivity of the assay was 0.4 pg/well, which corresponded to 20 pg/ml plasma. Plasma volumes for the assay ranging from 10 to 50 microl did not influence the PGFM standard curve; however, a slight drop in the OD450 was observed with higher plasma volumes. Biological validation of the assay was carried out in buffalo plasma samples obtained during physiological states of cyclicity, peri-estrus, post-insemination, reproductive tract infection and persistent corpus luteum conditions. A pulsatile pattern of plasma PGFM release was observed prior to estrus when PGFM was determined in blood samples collected at hourly intervals of time. The PGFM pulsatility was not observed when blood sampling frequency of either 4 or 12 h was considered. The PGFM levels stayed high in peripheral circulation of buffaloes with reproductive tract infections and remained low throughout the sampling period in buffaloes having persistent corpus luteum. After an initial increase post-insemination, the plasma PGFM levels showed minor fluctuations. The assay was found to be sufficiently reliable and specific for estimation of PGFM levels in buffaloes. The standardization and validation of PGFM assay in buffalo opens the prospects of using PGFM levels as an indicator for reproductive health status monitoring in this species. PMID- 12753782 TI - Response of the 1-5 day-aged ovine corpus luteum to prostaglandin F2alpha. AB - The hypothesis that, in the ewe, prostaglandin (PG) F2alpha administration on day 3 after ovulation is followed by luteolysis and ovulation was tested using 24 animals. The ewes were treated with a dose of a PGF2alpha analogue (delprostenate, 160 microg) on days 1 (n=8), 3 (n=8) or 5 (n=8) after ovulation, was established by transrectal ultrasonography. Daily scanning and blood sampling were performed to determine ovarian changes and progesterone serum concentrations by radioinmunoassay. The treatment induced a sharp decrease of progesterone concentrations followed by oestrus and ovulation in all ewes treated on days 3 and 5 and in one ewe treated on day 1 (8/8, 8/8, 1/8; P<0.05). Seven ewes treated on day 1 did not respond to PGF2alpha treatment and had an inter-ovulatory cycle of normal length (17.4 +/- 0.5 days). However, the profile of progesterone concentrations during the cycle of these ewes was delayed 1 day (P<0.05) compared with a control cycle. The overall interval between PGF2alpha and oestrus for the 17 responding ewes was 42.4 +/- 2.3 h. In 15 of these ewes the ovulatory follicle was originated from the first follicular wave and the ovulation occurred at 60.8 +/- 1.8 h after PGF2alpha treatment. The other two responding ewes ovulated an ovulatory follicle originated from the second follicular wave between 72 and 96 h after treatment. These results support the hypothesis and suggest that refractoriness to PGF2alpha of the recently formed corpus luteum (CL) may be restricted to the first 1-2 days post-ovulation. PMID- 12753783 TI - The effect of oestradiol on postpartum uterine involution in sheep. AB - The present study tested the hypothesis that ovarian oestradiol increases the rate of uterine involution after parturition in sheep. The day after parturition, ewes were randomly assigned as un-operated controls (n=5), or a 3 cm silastic implant containing oestradiol (n=8) or empty (n=7) was sutured within the bursa of the ovary ipsilateral to the previously gravid uterine horn. Blood samples were collected daily for measurement of PGFM and acute phase proteins until 17 days postpartum when the ewes were slaughtered and the genital tract was collected. There was no consistent effect of treatment group on uterine involution determined by the collagen density, dry matter content, width, length, or weight of the genital tract. Furthermore, there was no evidence of a localised effect of oestradiol on involution as there were no significant differences between the previously gravid and non-gravid uterine horns. However, oestradiol treated ewes had lower plasma concentrations of 15-keto-13,14-dihydro prostaglandin F2alpha (P<0.01), alpha1-acid glycoprotein (P<0.05) and ceruloplasmin (P<0.001); but, not haptoglobin. These observations could reflect a direct effect of oestradiol on inflammatory mediator synthesis or secretion because, in the absence of parallel physical measurements, it is unlikely that these observations reflect a beneficial effect of treatment on uterine health. PMID- 12753784 TI - The regulation of steroidogenesis by opioid peptides in porcine theca cells. AB - The present study was designed to investigate basal and LH-induced steroidogenesis in porcine theca cells from large follicles in response to various concentrations (1-1000 nM) of mu opioid receptor agonists (beta endorphin, DAMGO, FK 33-824), delta receptor agonists (met-enkephalin, leu enkephalin, DPLPE) and kappa receptor agonists (dynorphin A, dynorphin B, U 50488). Agonists of mu opioid receptors suppressed basal androstenedione (A4), testosterone (T) and oestradiol-17beta (E2) secretion and enhanced LH-induced A4 and T release by theca cells. The inhibitory effect of the agonists on E2 secretion was abolished in the presence of LH. All delta receptor agonists depressed basal progesterone (P4) output. However, the influence of these agents on LH-treated cells was negligible. Among delta receptor agonist used only leu enkephalin and DPLPE at the lowest concentrations inhibited basal A4 release. The presence of LH in culture media changed the influence of these opioids from inhibitory to stimulatory. Similarly, DPLPE reduced T secretion by non-stimulated theca cells and enhanced T secretion of stimulated cells. All of delta agonists inhibited basal E2 secretion and unaffected its release from LH-treated theca cells. Agonists of kappa receptors inhibited basal, non-stimulated, P4 secretion and two of them (dynorphin B, U 50488) potentiated LH-induced P4 output. Basal A4 and T release remained unaffected by kappa agonist treatment, but the cells cultured in the presence of LH generally increased both androgen production in response to these opioids. Basal secretion of E2 was also suppressed by kappa agonists. This inhibitory effect was not observed when the cells were additionally treated with LH. In view of these findings we suggest that opioid peptides derived from three major opioid precursors may directly participate in the regulation of porcine theca cell steroidogenesis. PMID- 12753785 TI - Antioxidant supplementation in vitro improves boar sperm motility and mitochondrial membrane potential after cryopreservation of different fractions of the ejaculate. AB - Antioxidant supplementation during cooling was assayed to improve the motility of frozen-thawed (FT) boar spermatozoa from two different fractions of the ejaculate, the first component of the sperm-rich fraction (Fraction I) and the rest of the bulk ejaculate (Fraction II). Using a split-sample design, addition of two different concentrations (100 and 200 microMl(-1)) of the water-soluble Vitamin E analogue Trolox (6-hydroxy -2,5,7,8-tetramethylchroman -2-carboxylic acid) was evaluated for an effect on sperm motility (measured both subjectively and by means of a computer assisted motility assessment (CASA)), and on mitochondrial membrane potential using flow cytometry after cell-loading with JC 1. The effect of the Vitamin E analogue was clearly dose-dependent and varied with the fraction of the ejaculate considered. Motility was significantly higher in Trolox-treated spermatozoa (200 microm), from either ejaculate fraction, albeit the effect was more evident in spermatozoa from Fraction II (P<0.05) for any Trolox-concentration. Antioxidant supplementation resulted, also dose dependent, in a higher number of spermatozoa showing high mitochondrial activity as assessed by the JC-1 staining, in both ejaculate fractions. In the present trial, exogenous Trolox positively affected post-thaw sperm viability (as motility and mitochondrial membrane potential) in both fractions of the ejaculate. The magnitude of the effect appeared, however, to be dependent of the fraction of the ejaculate considered. PMID- 12753787 TI - Influence of short-term relocation and male exposure on sexual receptivity and reproduction in artificially inseminated lactating doe rabbits. AB - A study was conducted to evaluate the effect of short-term relocation and male exposure on receptivity rate, kindling rate and total born per litter in lactating does under an artificial insemination (AI) programme. Thirty-two, 2 month-old New Zealand White rabbits were randomly allocated to one of four treatments: (1) relocation and male exposure; (2) relocation without male exposure; (3) no relocation with male exposure; (4) no relocation without male exposure (control). Relocation and male exposure were done 8-10 h before the time of service. First insemination was when does reached 3200 g body weight and does were bred 4-13 days after parturition across parities during a 6 month reproduction period. Of all breeding records, 125 inseminations and 91 kindlings were from nursing does. The mean interval from parturition to insemination for nursing does was 10.3 days. Relocation of lactating does resulted in greater (P<0.01) receptivity rate at service (74.8%) as compared with no relocation (55%). Receptivity rate was not influenced by male exposure. However, the interaction of relocationxmale exposure tended to be significant (P=0.07). Receptivity rate in relocated does exposed to males was 62.8 and 86.7% without exposure while in non-relocated does male exposure showed no effect. Kindling rate was not influenced by relocation or male exposure. The mean total born per litter in relocated and non-relocated does was 8.05 +/- 0.33 and 7.39 +/- 0.36, respectively, but no significant difference was observed. There was no effect of male exposure on total born per litter (7.85 +/- 0.34 versus 7.59 +/- 0.34 without male exposure). However, interaction of relocationxmale exposure on this variable was significant (P=0.009). Male exposure in relocated does decreased the size of the litter (7.52 +/- 0.46 versus 8.58 +/- 0.47 without male exposure) whereas mean values in non-relocated does increased when they were exposed to males (8.18 +/- 0.52 versus 6.60 +/- 0.49). Short-term relocation improved receptivity rate and reproduction in lactating does under an artificial insemination programme. Preliminary results indicated that male exposure in non relocated does improves the total born per litter at a similar level than relocated does without male exposure. Relocation combined with male exposure decreased receptivity rate and total born per litter as compared with relocated does without male exposure, but the reproductive performance in the former was greater as compared with those does where no relocation occurred without male exposure. PMID- 12753786 TI - Role of estradiol-17beta on nuclear and cytoplasmic maturation of pig oocytes. AB - The role of estradiol-17beta on nuclear and cytoplasmic maturation of pig oocytes was investigated in the present study. To determine the estradiol effect, oocytes were cultured for 42 h in a steroid free medium composed of mTCM-199 supplemented with LH, FSH and 10% charcoal extracted follicular fluid. Estradiol receptor (ER), detected by a binding assay, were present in cumulus cells and oocytes during maturation with higher levels observed at 24 h of culture in the oocytes and at 36 h in the cumulus cells. To block estradiol action an antiestrogen (1-p dimethylaminoethoxyphenyl-1,2-diphenyl-1-butene (tamoxifen)) was added to the maturation medium at various concentrations. The percentage of treated oocytes that underwent nuclear maturation was similar (P>0.05) to the control group. Cytoplasmic maturation, determined by the ability to form female pronucleus (FPN) and male pronucleus (MPN), was not different (P>0.05) among all groups. The presence of 4-hydroxy-4-androstene-3-17-dione (4-OHA) also did not influence nuclear (P>0.05) or cytoplasmic maturation (P>0.05). The results suggest that estradiol is not involved in maturation of pig oocytes. However, the present experiment used pronuclei formation as the endpoint, no studies were done in regard to estradiol's effects on the embryonic development. PMID- 12753788 TI - Intracellular adenosine triphosphate and glutathione concentrations in oocytes from first estrous, multi-estrous, and testosterone-treated gilts. AB - Cytoplasmic maturation refers to a variety of cellular changes that must occur in the oocyte in order to progress through subsequent fertilization and embryonic development. Intracellular concentrations of ATP (ATPi) or glutathione (GSHi), indicative of metabolic activity or the ability of the oocyte to form a male pronucleus and cope with cellular stress, respectively, have been used as markers of cytoplasmic maturation in vitro. In the current study, our objective was to determine if concentrations of ATPi and GSHi in oocytes recovered from three groups of gilts were associated with known differences in developmental competence within these populations. In vivo matured oocytes were surgically recovered 36-38 h after the onset of estrus from first estrous gilts, multi estrous gilts, and multi-estrous gilts receiving testosterone (1mg/2 ml per day; day 13 to estrus, day 0=day of estrus). Concentrations of ATPi and GSHi were determined using a bioluminescent somatic cell assay kit (luciferin-luciferase reaction) and the dithiobisnitrobenzoic acid (DTNB)-glutathione reductase recycling reaction, respectively. There were no differences (P>0.05) between ATPi concentrations in oocytes from the three groups (1.52 +/- 0.10, 1.51 +/- 0.11, 1.56 +/- 0.11pmol per oocyte). In contrast, oocytes from multi-estrous gilts had higher (P<0.05) concentrations of GSHi (31.53 +/- 1.66 to 33.67 +/- 2.30 pmol per oocyte) than oocytes from first estrous gilts (25.07 +/- 0.82). Administration of testosterone did not affect (P>0.05) GSHi concentrations in oocytes from multi estrous gilts. Differences in developmental potential between the three groups of gilts were apparently not due to different concentrations of ATPi. However, GSHi concentrations were higher in oocytes from multi-estrous gilts, suggesting that reduced developmental potential of oocytes from first-estrus gilts may be related to insufficient amounts of GSHi. The beneficial effect of exogenous testosterone on the percentage of embryos surviving early gestation does not appear to be due to increased GSHi. Of the numerous potential markers of developmental potential, two were examined in the current study, and GSHi appeared to be useful for assessing porcine oocytes. PMID- 12753789 TI - "If you want to sit on your butts you'll get nothing!" Community activism in response to threats of rural hospital closure in southern New Zealand. AB - Problems of ensuring rural health services in New Zealand have intensified as successive governments have attempted to limit expenditure and health agencies have seen rural services as relatively 'expendable'. From the literature two sets of indicators are identified: the factors influencing successful retention of rural services and the outcomes for the community. Interview and documentation data indicate the extent to which these characteristics and outcomes were present in nine rural community health trusts in southern New Zealand during the 1990s. Variability in outcome and success of community response to threats to rural services relates to the factors identified from the literature, particularly community leadership and capability, but also the prominent role played by local professionals. Given the common political and economic context, these local factors proved important in determining which communities successfully retained hospital services. PMID- 12753790 TI - The Aral Sea disaster and self-rated health. AB - This study examined the effect of psychosocial factors and environmental perceptions on self-rated health in the environmentally devastated Aral Sea area of Karakalpakstan. Self-rated health was assessed using a questionnaire on 881 randomly selected individuals from three communities. Communities were chosen based on relative differences with regards to economic and ethnic characteristics, and distance from the sea coast. Consistent with mortality rates in the area, the prevalence of 'poor' self-rated health was high. Factors negatively associated with self-rated health include psychosocial impacts and reported environmental concern, as well as community of residence and age. These results demonstrate that the population has a poor perception of their own health, a significant finding given that self-rated health is a strong predictor of morbidity and mortality. It is also clear that psychosocial health is strongly associated with health perceptions. Thus, to improve the overall health of this population, health remediation measures must address physical as well as psychosocial health problems. PMID- 12753791 TI - Therapeutic landscapes and First Nations peoples: an exploration of culture, health and place. AB - This paper contributes to an expanding body of research within Health Geography that focuses on the role of therapeutic landscapes in shaping health. Therapeutic landscapes demonstrate the importance of places for maintaining physical, emotional, mental and spiritual health. Meanings of place and the relationship between place and health have culturally specific dimensions, yet these tend to be overlooked especially with respect to First Nations peoples. This paper broadens the analysis of therapeutic landscapes by exploring their culturally specific dimensions in the context of the everyday lives of 'Anishinabek' and thus contributes to a better understanding of First Nations peoples. First Nations peoples contend that the relationship they have with the land shapes the cultural, spiritual, emotional, physical and social lives of individuals and communities. While geographic research has explored First Nations peoples health, few studies have attempted to explore the influence of cultural beliefs and values on health-let alone the intricate link between the land and health. This paper presents the results of 17 in-depth interviews conducted with Anishinabek (Ojibway and Odawa) living in one First Nations community in northern Ontario, Canada. The findings from the interviews demonstrate that culture is an important component of the link between health and place in everyday life. Incorporating First Nations peoples' perspectives of health and place reveals that the current conceptualizations of health and place within the Geography of Health literature are only partial. PMID- 12753792 TI - "It's your whole way of life really": negotiating work, health and gender. AB - Recent discussions in contemporary geographies of health have highlighted the need to understand the social contexts in which people experience health and illness. Qualitative and mixed method studies have been shown to be invaluable to such research, especially where investigations seek to understand the circumstances and responses surrounding particular conditions. In this paper, we move beyond biomedical approaches to combine methods in health research and gain insights into the complex contexts and relations affecting men's and women's respiratory health. Drawing on three literatures and past work on respiratory disease in selected primary industries, we report on qualitative research conducted with men and women horse trainers and vegetable growers working in southern New Zealand. We note the gendered work differences that could be affecting contrasting disease rates and focus on notions of 'metaphor' and 'place' to analyse the narratives trainers and growers construct about their health. We report that men and women negotiate a "whole way of life" that involves both specific workplaces and social relations that shape their work and health experiences. PMID- 12753793 TI - Emerging new research in the geography of health and impairment. PMID- 12753794 TI - Public honour, private shame and HIV: issues affecting sexual health service delivery in London's South Asian communities. AB - Within a wider discussion of health service provision for black and minority ethnic (BME) groups, this paper considers how socio-cultural factors affect the provision of HIV and sexual health services to South Asians in London. It argues that communally held concepts of honour and shame within South Asian communities create a framework of social control with significant implications for HIV/AIDS transmission. It examines the provision of culturally sensitive services to BME communities by ethnically specific and generic service providers through a case study of the Naz Project London. Finally, it proposes an agenda for future research into BME sexual health service provision. PMID- 12753795 TI - (Dis)abling children in primary school micro-spaces: geographies of inclusion and exclusion. AB - The geography of disabled children's schooling in the United Kingdom (UK) is changing, and this is underpinned by a growing international consensus that disabled children should be educated within mainstream school settings (UNESCO, The Salamanca statement and framework for action on special educational needs. World Conference on Special Needs Equality and Quality, Salamanca, Spain, 1994). As a result, new geographies of desegregation in disabled children's education are emerging, with disabled children being increasingly educated within mainstream rather than 'special' schools. This paper explores this issue, focusing on the (re)production of discourses of 'inclusion' and 'disability' in two mainstream primary schools in England. Empirical findings demonstrate that school actors reproduce meanings of inclusion and disability in different ways within and between school settings. It is shown that discourses of inclusion are frequently based on educational-medical models of disability, and can serve to exclude some children from mainstream schools. With this in mind, the paper highlights the value of a spatially sensitive evaluation of inclusion, that emphasises the importance of schools as unique moments in space and time to everyday practices of inclusion and disability. PMID- 12753796 TI - "Nae as nice a scheme as it used to be": lay accounts of neighbourhood incivilities and well-being. AB - This paper is based upon qualitative research conducted in a relatively deprived neighbourhood in Edinburgh, Scotland, and it explores how contextual and compositional features of place may interact to influence well-being. I present women's accounts of their experiences of neighbourhood incivilities, and I discuss their perceptions of the influence of incivilities upon their sense of well-being. I suggest that aspects of individual biographies mediate experiences of neighbourhood incivilities. Furthermore, some individuals seem to engage in "distancing strategies", which I suggest may be interpreted as a way of resisting the potentially harmful psycho-social influence of incivilities upon well-being. PMID- 12753797 TI - Predicting smoking behaviour in census output areas across Scotland. AB - Spatially disaggregated surveys of smoking behaviour are rare and hence estimating the geography of the incidence of smoking is difficult. The main aim of this study is to develop a technique for estimating smoking probability for different age/sex groups in small areas across the whole of Scotland using information on smoking behaviour from the Scottish Household Survey. This is useful not only in its own right, but as an aid to studies of geographical variations in diseases such as lung cancer that, as a first step, need to control for smoking behaviour. The method developed uses individual-level characteristics from the Scottish Household Survey combined with a set of output area and pseudo postcode sector measures from the 1991 census to model the probability of smoking. The parameters from this model are then used to make smoking predictions by age and sex for output areas across Scotland. This is the first time that such geographically detailed estimates of smoking have been made available. PMID- 12753798 TI - Reading maps of the genes: interpreting the spatiality of genetic knowledge. AB - Genetics has become the pre-eminent interpretation of the body and health and illness. This paper engages with a central technique and metaphor of the new genetics-gene mapping. Through an exploration of the process of gene mapping, the paper argues that the genetic material of the body is spatialised and transformed into a knowable and manipulable entity. Three interpretations of this spatial transformation of the body's materiality are discussed, in turn drawing on Foucault's notion of the construction of medical knowledge, the deconstruction of geographical maps and Haraway's 'fetishised' conception of the gene map. The paper concludes by considering contestations to this dominant discourse, and begins to construct an alternative spatialisation of the body that attempts to 'place' the gene more appropriately in a socially-embedded body and health. PMID- 12753799 TI - "Constant Medical Supervision": locating reproductive bodies in Victorian and Edwardian Dundee. AB - The literature on infant mortality has remained largely detached from the burgeoning literature on the body and embodiment. This paper reconsiders these two literatures in relation to the dynamics of industrialisation, social reform, and struggles over women's bodies in late 19th and early 20th century Dundee. During this period, Dundee was dominated by the jute industry which employed a largely female workforce. This industrial nexus was articulated within a broader environment of social improvement that was preoccupied with population growth and national power. As the need for healthy 'future citizens' became imperative, Dundee's infant mortality rate became a crucial marker in assessing working women's role and care of self. This paper pays specific attention to the strategies of reform that tracked, monitored and disciplined these working women and their 'reproductive' bodies. It focuses on the forms of knowledge and systems of knowledge production that characterised these projects, and how the conjunctures between power and knowledge were grounded in material spatial practices, transforming women's social and physical place in Dundee, and forming an urban geography of 'health reform'. PMID- 12753801 TI - How important is GAD in the etiology of spontaneous disease in human and murine type 1 diabetes? PMID- 12753802 TI - Conversations with GAD. PMID- 12753803 TI - GAD-about BDC2.5: peptides that stimulate BDC2.5 T cells and inhibit IDDM. PMID- 12753804 TI - Large antigens such as GAD-65 become involved in pathogenic self-reactivity, but fortunately only rarely. PMID- 12753805 TI - Antigen-induced T1D in NOD mice. PMID- 12753806 TI - Induction of autoimmune valvulitis in Lewis rats following immunization with peptides from the conserved region of group A streptococcal M protein. AB - Rheumatic heart disease (RHD) is considered to be an autoimmune disorder mediated by group A streptococcal (GAS) M protein-specific T cells and antibodies that cross-react with cardiac antigens and epitopes of the GAS M protein. In this study, Lewis rats were immunized with a pool of overlapping peptides spanning the conserved region of the GAS M protein in Complete Freund's Adjuvant, followed by immunization with Bordetella pertussis. Controls received adjuvants alone. Spleen derived lymphocytes from rats immunized with the conserved region peptides proliferated in response to the immunogen and to cardiac myosin. Moreover, histological examination of cardiac tissue from rats immunized with conserved region peptides revealed the presence of inflammatory lesions in both the myocardium and valve tissue indicating a role for GAS M protein-specific autoreactive T cells in the development of cardiac lesions. This study may support the use of the rat model of autoimmune valvulitis to investigate the immunopathogenesis of RHD and possible preventive strategies. PMID- 12753807 TI - Defect in activation-induced cell death in non-obese diabetic (NOD) T lymphocytes. AB - Activation-induced cell death (AICD) represents a major means of peripheral tolerance induction, eliminating effector cells. NOD mice, a widely used model for autoimmune diabetes, are characterized by high levels of circulating T lymphocytes and by resistance to several apoptosis-inducing signals. The aim of this study was to analyse AICD in peripheral NOD T lymphocytes. First, we demonstrated in an in vitro AICD model that NOD T lymphocytes are more resistant to AICD (64+/-2%) compared to non-autoimmune C57BL/6 T lymphocytes (73+/-2%), but also diabetes-resistant NOR T lymphocytes (76+/-3%, P<0.05). Moreover, both CD4(+)and CD8(+)subsets were affected. Analysis of the cellular and molecular pathways revealed lower caspase 8 levels, a central caspase proximally involved in the AICD-pathway (fluorescence of 258+/-47 in NOD vs. 441+/-16 in NOR and 414+/-61 in C57BL/6 T lymphocytes, P<0.05). Gene expression analysis using real time RT-PCR additionally revealed low expression of Fas and FasL, the death receptor system activating caspase 8 and contributing to AICD. Additionally, low IL-2 levels, together with high TGFbeta and Bclx-L levels, confirm the presence of a NOD-specific AICD-resistance profile. In conclusion, we present cellular and molecular evidence for disturbed AICD mechanisms in NOD T lymphocytes. This resistance in AICD may contribute to defective tolerance induction to autoantigens in NOD mice. PMID- 12753808 TI - Deregulation of peripheral B-cell development in enhanced severity of collagen induced arthritis in FcgammaRIIB-deficient mice. AB - Accumulating evidence indicates that the type IIB Fc receptor for IgG (FcgammaRIIB) plays a pivotal role in maintaining peripheral tolerance by suppressing excessive humoral and cellular immune responses. However, little is known about the mechanism by which the autoreactive B cells develop in the periphery in FcgammaRIIB-deficient mice. To clarify the role of FcgammaRIIB in the emergence of autoreactive B cells, we analyzed B-cell compartments in the autoimmune arthritis-susceptible DBA/1 mice devoid of FcgammaRIIB (DBA.IIB-/-) during the induction of collagen-induced arthritis (CIA). We found that DBA.IIB-/ showed an increase in the number of peripheral immature type 2 transitional (T2) B cells after immunization with type II collagen (C-II), followed by the enhanced severity of CIA with higher autoantibody titers to mouse C-II than those of wild type DBA/1. In addition, elevated secretion of IL-1alpha by peritoneal macrophages from DBA.IIB-/- on stimulation with IgG immune complexes in vitro suggested the augmented effector cell responses in the CIA course of DBA.IIB-/-. These findings suggest that the FcgammaRIIB-dependent triple regulation in the peripheral T2 B cells, in the antibody production, and in the effector cell responses is crucial for suppressing CIA. PMID- 12753809 TI - Type 1 diabetes alters anti-hsp90 autoantibody isotype. AB - The 90-kDa chaperon family includes heat shock protein (hsp) 90 and glucose regulated protein (grp) 94. These proteins play an important role in normal cellular architecture, in the etiology of some autoimmune and infectious diseases and in antigen presentation to T cells. Owing to its role in autoimmunity, we explored anti-hsp90 autoantibody (hsp90AA) response in the sera of persons with type 1 diabetes, first-degree relatives (FDR) and in normal subjects. Significant high level of hsp90AA was found in FDR, but there was no significant difference between the normal and diabetic persons. The IgG1 and IgG3 isotypes of hsp90AA were higher in persons with type 1 diabetes and FDR than in normal subjects. We found a good correlation between hsp90AA measured by ELISA and RIA. A positive correlation between serum hsp90AA and glutamic acid decarboxylase (GAD65) autoantibody (GAA) was also observed. Hsp90AA positive sera from diabetic persons immunoblotted recombinant hsp90, GAD65 and corresponding proteins in islet lysates. Our study suggests that hsp90AA are present in normal, FDR and diabetic persons. However, there is a higher level of IgG1 and IgG3 isotypes of hsp90AA in FDR and type 1 diabetic subjects. Thus, autoimmunity leading to type 1 diabetes significantly alters the autoantibody isotype to autoantigens, such as hsp90. PMID- 12753810 TI - Profile and clinical significance of anti-nuclear envelope antibodies found in patients with primary biliary cirrhosis: a multicenter study. AB - Primary biliary cirrhosis (PBC) sera contain antibodies which recognize various nuclear envelope proteins of which antibody against gp210 has been proven to be diagnostic for disease. In contrast, the clinical significance of another nuclear envelope antibody, anti-p62 antibody has not been well investigated. In the present study, we have analyzed anti-nuclear envelope antibodies by indirect immunofluorescence and immunoblot using rat liver nuclear envelope proteins and wheat germ agglutinin-bound fraction. Test sera were obtained from 175 patients with PBC and from 120 controls. Anti-gp210, anti-lamina associated polypeptide 2, anti-lamin B receptor, and anti-p62 complex antibodies were detected with a frequency of 26% (46 of 175), 6% (11 of 175), 9% (16 of 175), and 13% (15 of 115), respectively. The confirmation of Scheuer's stage IV was made with a frequency of 27% (4 of 15) in PBC patients with anti-p62 complex antibody, in contrast to only 2% (2 of 100) in PBC patients without anti-p62 complex antibody. This difference was found to be statistically significant. The presence of anti p62 complex antibody may be related with the progressive or advanced state of PBC. PMID- 12753811 TI - Promiscuous T cells selected by Escherichia coli: OGDC-E2 in primary biliary cirrhosis. AB - The etiology of primary biliary cirrhosis (PBC) remains enigmatic. One theory that has attracted attention proposes that PBC is induced via molecular mimicry with Escherichia coli. If molecular mimicry is responsible for the immunogenic response in PBC, then T cell clones specific for E. coli antigens should stimulate and be cross-reactive with peptides specific for the human immunodominant autoepitopes. To address this issue, we developed T cell clones specific for E. coli OGDC-E2 peptide. Importantly, we demonstrate the presence of T cell clones specific for E. coli OGDC-E2 that react promiscuously with the human mitochondrial equivalents. Indeed, there was a significant increase in the liver derived T cell precursor frequency of such reactivity and such liver clones were only found in patients with PBC. In conclusion, these data suggest that PBC is a multi-hit disease involving a genetic predisposition, a mucosal response, and activation of promiscuous T cells; such activation may occur either directly from bacterial antigens, or indirectly through chemically-modified bacterial antigens. Dissection of the mechanisms involved will lead not only to understanding the immunogenetic basis of PBC, but likely its pathogenic etiology. PMID- 12753812 TI - Sustaining safe practice: twenty years on. AB - This paper examines the ways in which populations at risk of HIV in the developed world have enculturated the knowledges and technologies of both the medical and the social sciences. By revisiting a number of review papers and by reviewing findings from a range of studies, we argue that gay men have appropriated information that has enabled them to sustain safe practices while they have eschewed information that has made maintenance difficult. The paper describes a range of risk reduction strategies and compares the responses of populations at risk of HIV in the years before the advent of highly active antiviral therapy (HAART) with their responses after the introduction of HAART in 1996. We concentrate our argument on the changing responses to HIV risk of gay men, although occasionally illustrate our argument with reference to the responses of injecting drug users. The responses of gay men to risk post-HAART--particularly those who reside in Australia--speak to the adoption of a range of considered strategies, not altogether safe, to reduce harm. We argue that such strategies need to be understood and addressed within a 'new' social public health, that is, a public health that takes what social analysis has to say seriously. The paper examines the differences between the traditional, the 'modern' epidemiological/clinical and the 'new' social or socio-cultural public healths and describes the tensions between the medical and the social science disciplines in their efforts to inform public health. Key concepts provided by social science such as agency (including individual and collective agency), alongside its methodological reflexivity are key to effective public health. The risk avoidance strategies adopted by gay men suggest a way forward by turning our attention to the ways in which medicine is taken in(to) their practice. PMID- 12753813 TI - HIV and AIDS-related stigma and discrimination: a conceptual framework and implications for action. AB - Internationally, there has been a recent resurgence of interest in HIV and AIDS related stigma and discrimination, triggered at least in part by growing recognition that negative social responses to the epidemic remain pervasive even in seriously affected communities. Yet, rarely are existing notions of stigma and discrimination interrogated for their conceptual adequacy and their usefulness in leading to the design of effective programmes and interventions. Taking as its starting point, the classic formulation of stigma as a 'significantly discrediting' attribute, but moving beyond this to conceptualize stigma and stigmatization as intimately linked to the reproduction of social difference, this paper offers a new framework by which to understand HIV and AIDS-related stigma and its effects. It so doing, it highlights the manner in which stigma feeds upon, strengthens and reproduces existing inequalities of class, race, gender and sexuality. It highlights the limitations of individualistic modes of stigma alleviation and calls instead for new programmatic approaches in which the resistance of stigmatized individuals and communities is utilized as a resource for social change. PMID- 12753814 TI - Nuances and shifts in lesbian women's constructions of STI and HIV vulnerability. AB - This study examines the subjective side of vulnerability as a social construct rooted in interpersonal relationships and community membership. Analysis is based on a survey of an especially diverse sample of 162 lesbian women, 67 of whom also participated in depth interviews. Another 24 of the original sample also participated in transcribed focus groups. One third were African American, Latina, and Asian, and two thirds were white. This sample reported an overall infection rate of 23%. Three subjective stances, or risk frames, are identified: essentially invulnerable, socially inoculated, and fundamentally vulnerable. Some women describe shifts in their interpretations of their own vulnerability, moving from one stance to another in response to obtaining information, becoming infected, having friends or acquaintances who become infected, and becoming involved with new partners. It is suggested that these shifts comprise a subjective "vulnerability career". The significance of lesbian women's constructions of vulnerability is examined, and the implications of this study for a better understanding of their risk for STIs are discussed. PMID- 12753815 TI - Situational factors influencing drug injecting, risk reduction and syringe exchange in Togliatti City, Russian Federation: a qualitative study of micro risk environment. AB - We undertook a qualitative study to explore the micro-environment of drug injecting, risk reduction and syringe exchange practices among injecting drug users (IDUs) in Togliatti City, Russia. Semi-structured qualitative interviews (n=57) were undertaken with current IDUs in May 2001. Findings highlight a recent transition away from hanka (a home-produced liquid opiate derived from opium poppy) towards the injection of heroin powder, and a drug use culture in which injecting predominates. Findings emphasise that risk reduction practices may be influenced less by availability of injecting equipment than by an interplay of situational and micro-environmental factors. Principal among these is a reported fear of police detainment or arrest among IDUs which encourages a reluctance to carry needles and syringes, and which in turn, is associated with needle and syringe sharing at the point of drug sale. We note the role of policing practices in influencing risk reduction and the potential role of policing agencies in supporting HIV prevention initiatives among IDUs. PMID- 12753816 TI - A proper place to live: health inequalities, agency and the normative dimensions of space. AB - This paper explores the links between lay knowledge, place and health related social action (or agency) at the individual and collective level. It is based on an analysis of in-depth interviews and neighbourhood survey data across four localities in two cities in the North West of England.The qualitative analysis has identified 'guidelines' that we argue provide socially shared understandings of the normative contours of 'proper places' which shape the way people respond to the everyday lived reality of places. The quantitative findings suggest that a substantial minority of people, particularly in disadvantaged areas, are exposed to significant dissonance between the normative dimensions and lived experience of place. The analysis points to potential interactions between individual and collective action which may affect the health of individuals and populations and 'ontological fit'-people's ability to (re) construct a positive identity despite living in what they and others perceive to be an 'improper' place. This is linked to their biographical connections with particular places and the extent to which they can localise problems and people in places at a distance from themselves. The paper contributes to understanding about the processes that generate inequalities in the health experience of people living in sharply contrasting socio-economic circumstances as well as finer-grained health inequalities between the 'poor' and the 'poorest'. PMID- 12753817 TI - Measuring culture: a critical review of acculturation and health in Asian immigrant populations. AB - The number of studies examining how acculturation affects the health of Asian immigrants has increased in recent years. The proliferation of studies reflects the growing size and heterogeneity of Asian immigrant populations in the United States, Canada, Australia, New Zealand, and the United Kingdom. This paper compares various approaches to acculturation within the health literature on Asian immigrants by reviewing the literature in three-health domains (1) mental health (2) physical health and (3) health services use. The review critically examines the conceptualizations and measures of acculturation in these three domains and presents major findings. We observe that measurement difficulties posed by the experiences of heterogeneous Asian groups compound theoretical and disciplinary disparities between acculturation instruments. The extent to which conceptual and methodological critiques of acculturation studies in Hispanic populations apply to studies of Asian populations is also discussed. The critical review thus provides insights into the diverse ways that the relationship between culture and health is measured in this complicated and growing literature. PMID- 12753818 TI - Patients' attitudes vs. physicians' determination: implications for cesarean sections. AB - Most research studies identifying non-clinical factors that influence the choice of Cesarean Section as a method of obstetric delivery assume that the physician makes the decision. This paper arguably shows the role played by the mother. Owing to the fact that Chinese people generally believe that choosing the right days for certain life events, such as marriage, can change a person's fate into a better one, the hypothesis is tested that the probability of Cesarean Sections being performed is significantly higher on auspicious days and significantly lower on inauspicious days. By employing a logistic model and utilizing 1998 birth certificate data for Taiwan, we are able to show that the hypothesis is accepted. PMID- 12753819 TI - Trust, learning, and vaccination: a case study of a North Indian village. AB - For US 17 dollars a child can be immunized against six major illnesses. Even at this price, a country such as India would have to spend half its health budget on providing vaccinations. Given the wide variation in immunization costs it may be possible to decrease this cost to more sustainable levels, but to do so we need to arrive at a more thorough understanding of factors affecting household demand for vaccination. Using data on vaccination and pre-natal care collected by the authors in the Garhwal region of India, we explore one aspect of the demand for vaccination in some detail. We show that informational constraints play an important role in the household decision to seek vaccination, and moreover, that learning about the efficacy of vaccinations only through empirical observation may be hard even in environments with variation in vaccination and the high incidence of vaccine-preventable diseases. We argue that when learning about vaccination is inefficient, households use concurrent interventions with easily observable outcomes to evaluate the veracity of a provider's claim regarding preventive care. Hence, the success of immunization programs becomes crucially linked to the success of parallel programs by the same provider. PMID- 12753820 TI - Television documentary in New Zealand and the construction of doctors by lower socio-economic groups. AB - The medical profession remains central to the provision of health care and the treatment of illness within contemporary society. However, the image of doctors and the relationship of the profession with the public is contested. The public persona of doctors has been subjected to re-negotiation in recent years as a result of factors such as health care reforms, the increased autonomy of other health professionals, the rise of the health consumer, and well-publicised cases of medical misadventure. We argue that television viewing is one influential way through which images of medical doctors are socially negotiated. This paper explores the construction of doctors through an analysis of television health documentary coverage and the accounts of lower SES participants in New Zealand. It demonstrates how televised depictions of doctors are integrated into the lifeworlds of viewers. We show that multiple and often contradictory representations of doctors, within both television health coverage and the accounts of our participants, conflate the traditional characterization of the caring professional with more recently established characterizations such as the medical entrepreneur and the bungling quack. The result is a complex and contextually variable image of doctors that embodies tensions surrounding public anxiety over health care reform. Recourse to this more pluralistic image of doctors provides a way for participants to work through the dilemmas posed by reduced access to medical care and the uncertainties of medical treatment, while still maintaining support for universal access to medical care. PMID- 12753821 TI - Socio-economic and demographic variation in health and in its measures: the issue of reporting heterogeneity. AB - True health state is an unobservable concept. Researchers and practitioners now have access to a large variety of tools to measure the health state and health related quality of life by self-reports. Socio-demographic variation in these measures is usually interpreted as variation in health. However, building on several measures simultaneously (multiple indicators), true health might be better represented, so that socio-demographic variation in any indicator can be decomposed into variation in the estimated true health, and measure-specific variation, holding true health constant. The latter variation is referred to as "reporting heterogeneity". Using structural equations models, the paper provides an empirical assessment of reporting heterogeneity in three popular measures of health and health related quality of life: the number of chronic conditions (CHRON), the SF-36 instrument and the visual analogue rating scale. Considering a large array of socio-economic and demographic characteristics from an Israeli health survey, the results indicate the existence of age-related reporting heterogeneity in the CHRON; income-related heterogeneity in the rating scale measure; and age, sex, income, ethnic origin and religiosity-related reporting heterogeneity in the SF-36 tool, in particular in its mental component scale. The main implication of reporting heterogeneity on the common uses of self-reported health measures is the need to adjust the measures not only for the determinants of health but also for the determinants of reporting heterogeneity. PMID- 12753822 TI - "I'll give up smoking when you get me better": patients' resistance to attempts to problematise smoking in general practice (GP) consultations. AB - This paper focuses on general practice consultations where the issue of patients' smoking is raised and problematised by general practitioners (GPs) by linking it to their presenting or ongoing medical conditions. The data are taken from a larger study in one country of the UK, of the factors influencing discussion of smoking between GPs and patients who smoke. Consultations have been examined informed by the conversation analysis literature, with a focus on patients' resistance to doctors' problematisation of smoking. It is argued that, despite evidence from other areas of health care that advice is most effective when it is personalised, and despite GPs' expressed views that a preferred way of topicalising smoking is to make links to a patient's current medical problems, this is not generally the case in these consultations. Linking smoking to current problems commonly results in explicit resistance from patients of a kind that is rarely seen in other medical consultations. It is postulated that this results from the moral implications of linking a person's health status with their own behaviour, thereby undermining their claim to legitimate illness and to medical help. PMID- 12753823 TI - Layperson and physician perceptions of the malpractice system: implications for patient safety. AB - The malpractice tort system functions upon the assumption that the medical profession defines its own standard of care. Hence, clinical assessments should theoretically mirror legal ones. However, if there is a conflict between the two, this conflict may reflect a perceived bias of the system either for or against a party. This exploratory study attempts to determine whether such a bias could exist. Physicians and layperson jury pool members were asked to review 10 jury verdict case scenarios. Respondents were asked first to assess whether the defendant physician provided clinically appropriate care; they were then asked to predict what the jury in the case actually decided. Laypersons showed significantly better agreement with actual jury verdicts on clinical assessment and success in jury verdict prediction than physicians. Both physicians and laypersons switched the favored party from clinical assessment to verdict prediction, with a vast majority of these changes being made from defendant to plaintiff. These results were consistent overall and when parsing assessments by case verdicts. Thus, laypersons and physicians may perceive a similar bias toward plaintiffs in the malpractice system. If these results can be generalized, the malpractice system may be inducing behavior that has a negative impact on patient safety. PMID- 12753824 TI - Willingness to pay and determinants of choice for improved malaria treatment in rural Nepal. AB - A logit model is used to estimate provider choice from six types by malaria patients in rural Nepal. Patient characteristics that influence choice include travel costs, income category, household size, gender, and severity of malaria. Income effects are introduced by assuming the marginal utility of money is a step function of expenditures on the numeraire. This method of incorporating income effects is ideally suited for situations when exact income data is not available. Significant provider characteristics include wait time for treatment and wait time for laboratory results. Household willingness to pay (wtp) is estimated for increasing the number of providers and for providing more sites with blood testing capabilities. Wtp estimates vary significantly across households and allow one to assess how much different households would benefit or lose under different government proposals. PMID- 12753825 TI - The impact on breastfeeding of labour market policy and practice in Ireland, Sweden, and the USA. AB - In recent decades there has been a marked rise in the labour market participation of women with infants in many countries. Partly in response to this trend, there are calls for greater emphasis on infant and child health in research and policy development on parental leave and other work-family balancing measures. Yet achieving high rates of breastfeeding as a health objective has thus far received relatively little attention in this context. Biomedical literature outlines the important health benefits conferred by breastfeeding, including upon infants and young children among middle class populations in developed countries. International recommendations now advise exclusive breastfeeding for 6 months. However, research indicates that the timing of the mother's resumption of employment is a key factor influencing the duration of exclusive breastfeeding. There would thus appear to be considerable potential for labour policy and practice, particularly maternity/parental leave provisions, to positively influence breastfeeding practice. Taking the case studies of Ireland, Sweden, and the United States, this paper explores the implications of labour market and early childhood policy for breastfeeding practice. The equity tensions posed by the breastfeeding-maternal employment intersection are also examined. The paper concludes that both socio-cultural support and labour market/health/early childhood policy are important if high rates of both breastfeeding and women's employment are to be achieved in industrialised countries. PMID- 12753826 TI - The politics of 'branding' in policy transfer: the case of DOTS for tuberculosis control. AB - How and why policies are transferred between countries has attracted considerable interest from scholars of public policy over the last decade. This paper, based on a larger study, sets out to explore the processes involved in policy transfer between international and national levels. These processes are illustrated by looking at a particular public health policy--DOTS for the control and treatment of tuberculosis. The paper demonstrates how, after a long period of neglect, resources were mobilised to put tuberculosis back on international and national public policy agendas, and then how the policy was 'branded' and marketed as DOTS, and transferred to low and middle income countries. It focuses specifically on international agenda setting and policy formulation, and the role played by international organisations in those processes. It shows that policy communities, and particular individuals within them, may take political rather than technical positions in these processes, which can result in considerable contestation. The paper ends by suggesting that while it is possible to raise the profile of a policy dramatically through branding and marketing, success also depends on external events providing windows of opportunity for action. Second, it warns that simplifying policy approaches to 'one-size-fits-all' carries inherent risks, and can be perceived to harm locally appropriate programmes. Third, top-down internationally driven policy changes may lead to apparent policy transfer, but not necessarily to successfully implemented programmes. PMID- 12753831 TI - Environmental chemistry of phosphonates. AB - Phosphonates are anthropogenic complexing agents containing one or more C PO(OH)(2) groups. They are used in numerous technical and industrial applications as chelating agents and scale inhibitors. Phosphonates have properties that differentiate them from other chelating agents and that greatly affect their environmental behavior. Phosphonates have a very strong interaction with surfaces, which results in a significant removal in technical and natural systems. Due to this strong adsorption, little or no remobilization of metals is expected. No biodegradation of phosphonates during water treatment is observed but photodegradation of the Fe(III)-complexes is rapid. Aminopolyphosphonates are also rapidly oxidized in the presence of Mn(II) and oxygen and stable breakdown products are formed that have been detected in wastewater. The lack of information about phosphonates in the environment is linked to analytical problems of their determination at trace concentrations in natural waters. Further method development is urgently needed in this area, including speciation of these compounds. With the current knowledge on speciation, we can conclude that phosphonates are mainly present as Ca and Mg-complexes in natural waters and therefore do not affect metal speciation or transport. PMID- 12753832 TI - Short-term harmful effects of unionised ammonia on natural populations of Moina micrura and Brachionus rubens in a deep waste treatment pond. AB - Populations of Moina micrura and Brachionus rubens in a deep waste treatment pond were exposed to the natural short-term fluctuations of unionised ammonia (90-min intervals of monitoring) that occur in the course of a day during a summer algal bloom. Under natural conditions, three replicate experiments were conducted in which water temperature, pH, dissolved oxygen, total ammonia, unionised ammonia, phytoplankton biomass and zooplankton (number of living and dead organisms, mortality rate and instant mortality) were studied. The time-course of unionised ammonia concentration was consistent with those shown by temperature, pH, phytoplankton biomass, dissolved oxygen, Moina micrura mortality and Brachionus rubens mortality. On the other hand, temperature, pH and dissolved oxygen never exceeded the tolerance ranges described for Moina and Brachionus, which led us to attribute the cause of zooplankton mortality to unionised ammonia toxicity. Mortality rates of 63%, 27% and 34% were recorded for Moina in each replicate experiment. Brachionus was less affected, with mortalities of 7.3%, 6.2% and 6.0%. These results confirm previous field observations (Water Res. 34(14) (2000) 3666; Water Res. 37(5) (2003) 1048) that attributed a reduction in zooplankton biomass during certain periods of summer (algal blooms) to a harmful side-effect of an excessive increase in phytoplankton biomass: high photosynthetic activity during these periods of proliferation of algae gives rise to an increased pH (>/=8) and, subsequently, leads to production of unionised ammonia (toxic for aquatic organisms) from its ionised fraction. PMID- 12753833 TI - Effect of feeding pattern and storage on the sludge settleability under aerobic conditions. AB - The selection of filamentous bacteria is often assumed to be associated with specific microbial properties such as growth rate, substrate uptake rate, substrate affinity and potential for substrate storage. In this study we aimed to verify some of these factors. Sequencing batch reactor (SBR) systems were used to scale-down aerobic activated sludge systems with an aerobic selector. Adding acetate in different aerobic feeding periods allowed us to simulate a variable relative size of aerobic selector with different bulk liquid substrate concentrations. The experiments showed that as expected, the aerobic fill time ratio (FTR(ox)) and the corresponding feast period, which can be assumed similar to contact time in an aerobic selector, had a strong effect on the sludge settleability. Promoting a strong substrate gradient in the SBR (FTR(ox)<5.4%) resulted in good sludge settleability (SVI<120mLg(-1)). Whenever acetate was added in a limiting rate (FTR(ox)>6.2%), a condition in which the acetate concentration in the reactor was always very low, the sludge settleability decreased (SVI>150mLg(-1)). Sludge settleability could be improved by changing the feeding strategy to a pulse feed. The maximum specific acetate uptake rate and poly beta-hydroxybutyrate (PHB) production rate of bad settling sludge, including bulking sludge, was similar to well-settling sludge, which is not in accordance with the general assumptions that well settling sludge have a higher maximal substrate uptake rate and better storage capacities. An alternative hypothesis for the development of filamentous structures in biological flocs has been formulated. It is hypothesized that bulking sludge originates from the presence of substrate gradients in sludge aggregates. Whereas at low bulk liquid substrate concentration filamentous bacteria give easier access to the substrate at the outside of the flocs and thereby proliferate, at high bulk liquid substrate concentration there is no substrate advantage for filamentous organisms and smooth bacterial structures predominate. In this hypothesis there is no need for an intrinsic difference in kinetic parameters between floc and filamentous bacteria. Where presence of filamentous bacteria is related to process conditions, the presence of a specific filament is likely due to presence of a specific limiting substrate. PMID- 12753834 TI - Urea hydrolysis and precipitation dynamics in a urine-collecting system. AB - Blockages caused by inorganic precipitates are a major problem of urine collecting systems. The trigger of precipitation is the hydrolysis of urea by bacterial urease. While the maximum amount of precipitates, i.e. the precipitation potential, can be estimated with equilibrium calculations, little is known about the dynamics of ureolysis and precipitation. To gain insight in these processes, we performed batch experiments with precipitated solids and stored urine from a urine-collecting system and later simulated the results with a computer model. We found that urease-active bacteria mainly grow in the pipes and are flushed into the collection tank. Both, bacteria and free urease, hydrolyse urea. Only few days are necessary for complete urea depletion in the collection tank. Two experiments with precipitated solids from the pipes showed that precipitation sets in soon after ureolysis has started. At the end of the experiments, 11% and 24% of urea was hydrolysed while the mass concentration of newly formed precipitates already corresponded to 87% and 97% of the precipitation potential, respectively. We could simulate ureolysis and precipitation with a computer model based on the surface dislocation approach. The simulations showed that struvite and octacalcium phosphate (OCP) are the precipitating minerals. While struvite precipitates already at low supersaturation, OCP precipitation starts not until a high level of supersaturation is reached. Since measurements and computer simulations show that hydroxyapatite (HAP) is the final calcium phosphate mineral in urine solutions, OCP is only a precursor phase which slowly transforms into HAP. PMID- 12753836 TI - Ozone decomposition of hazardous chemical substance in organic solvents. AB - Performance on the ozonation of hazardous chemicals in non-aqueous solutions, i.e. organic solvents, was studied in comparison with those in water. The specific conclusions obtained from this study are as follows: the rate of decomposition and the specific amount of decomposition per ozone consumption for orange II were higher in organic solvents like acetic acid, acetone, ter-butyl alcohol than in distilled water. The rates of trichloroethylene decomposition in organic solvents like acetic acid, acetone, and ethyl acetate were also higher than those in distilled water. PMID- 12753835 TI - Dynamic behavior of ozonation with pollutant in a countercurrent bubble column with oxygen mass transfer. AB - The dynamic behavior of ozonation with pollutants in a countercurrent bubble column is studied for the model establishment. Bubble columns have been widely used for an ozonation system in the plants and laboratories. In addition, a countercurrent bubble column has been commonly recommended than a cocurrent one because it has a higher ozone transfer efficiency. Therefore, the investigation of this paper focuses on the countercurrent bubble column. As an ozonation process starts, the gas mixture of ozone and oxygen is introduced into the bottom of a column, and then transferred into the liquid. The pollutants in the wastewater are eliminated subsequently via oxidation by the dissolved ozone. There certainly exists a temporary and unsteady period before the ozonation system reaches steady state. However, available ozonation models employed to describe ozone and pollutant profiles have commonly been developed for steady state. The treating qualities of wastewater in the early stage of ozonation are usually not predicted, and the time required for the steady-state establishment remains to be determined. Moreover, oxygen mass transfer is usually neglected in previous ozonation models so that the increase of dissolved oxygen is uncertain. These information is desirable for the proper design and operation of ozonation system in a bubble column. Thus, the aim of this study is to model and investigate the dynamic processes of ozonation with pollutants including oxygen mass transfer. The dynamic axial dispersion model proposed is employed to predict the variation of the ozone, pollutant, and oxygen concentrations profiles. The validity of the model was demonstrated by comparing the predicted results with the experimental data. The o-cresol was chosen as the model pollutant. The temporal concentration variations of the residual o-cresol and dissolved oxygen in the effluent liquid, and the off-gas ozone in the free volume were measured accordingly. Furthermore, the variation of the enhancement factor of ozone and the amount of off-gas were predicted. Note that it usually needs 2-5 hydraulic retention times to approach steady state under the conditions of this study. Further, the effects of dimensionless system parameters on the performance of the ozonation processes are examined. As a result, the proposed dynamic model of ozonation with pollutants is useful for proper prediction of the variables of an ozonation system in a countercurrent bubble column. PMID- 12753837 TI - Molecular characterization of dissolved organic matter in freshwater wetlands of the Florida Everglades. AB - In this study, the molecular composition of dissolved organic matter (DOM), collected from wetlands of the Southern Everglades, was examined using a variety of analytical techniques in order to characterize its sources and transformation in the environment. The methods applied for the characterization of DOM included fluorescence spectroscopy, solid state 13C CPMAS NMR spectroscopy, and pyrolysis GC/MS. The relative abundance of protein-like components and carbohydrates increased from the canal site to more remote freshwater marsh sites suggesting that significant amounts of non-humic DOM are autochthonously produced within the freshwater marshes, and are not exclusively introduced through canal inputs. Such in situ DOM production is important when considering how DOM from canals is processed and transported to downstream estuaries of Florida Bay. PMID- 12753838 TI - Molecular markers of anthropogenic activity in sediments of the Havel and Spree Rivers (Germany). AB - Detailed gas chromatographic/mass spectrometric analyses have been applied to sediment samples of the Havel and Spree River, tributaries to the Elbe River, in order to identify specific molecular markers of anthropogenic activities. Despite a wide variety of lipophilic organic compounds from diffuse anthropogenic contamination, a local emission of an industrial point source was reflected by specific markers including halogenated compounds and nitrogen containing substances (4-ethylnitrobenzene, formyl piperidine, acetyl piperidine). In addition to well-known anthropogenic markers various new molecular tracers were detected and are discussed, namely plasticizers (alkylsulfonic acid aryl esters, tributyl and tricresyl phosphates), synthetic fragrances (galaxolide, tonalide, 4 oxoisophorone), additives of personal care products (4-methoxycinnamic acid 2 ethylhexyl ester, benzyl benzoate, dibenzyl ether, benzophenone), occurring due to sewage treatment plant input. PMID- 12753839 TI - Removal of fish pathogenic bacteria in biological sand filters. AB - Documentation is required to evaluate the use of infiltration systems as an alternative method for removal of fish pathogenic bacteria in wastewater from fish-farms. This study was performed to investigate the removal of bacterial fish pathogens in biological sand filters. A second aim of the study was to evaluate the bacteria used in the study in order to find a suitable model organism for future experiments. Low-strength wastewater from an inland freshwater salmonid farm was intermittently loaded (70 mm/day in 24 doses) to filter columns containing either fine sand (d(10)=0.25) or coarse sand (d(10)=0.86). After a wastewater loading period of 10 weeks, separate sand columns were seeded with Yersinia ruckeri, Pseudomonas fluorescens, Aeromonas hydrophila and Aeromonas salmonicida subsp. salmonicida, respectively, for a period of 30 days. All the bacteria showed the same removal performance during the experiment, with a significantly lower removal in the beginning of the experiment (day 1-7) compared to mid- and late-phase (day 12-30). In mid- and late-phase the removal stabilized at a high level (>99.9%) for all the bacteria. The hydrophobic cell surface properties of the Aeromonads were higher than Ps. fluorescens and Y. ruckeri. This can possibly explain the significantly higher (P<0.05) removal efficiencies seen for A. hydrophila and A. salmonicida subsp. salmonicida compared to Y. ruckeri and Ps. fluorescens. Results were promising with regard to the use of low cost infiltration systems as an alternative disinfection method for fish-farm wastewater. Following the criteria for a suitable model organism (removal efficiency, detection in filter effluent and die-off in storage tanks), Y. ruckeri was found to be a feasible model organism for use in future experiments. PMID- 12753840 TI - Estimating DOC regime in a wastewater treatment plant by UV deconvolution. AB - A UV-deconvolution method was modified, and applied to estimation of dissolved organic carbon (DOC) along a municipal wastewater treatment plant (WWTP) in Catalonia, Spain. One grab sample was taken every 2h at four sampling points, for 1 week (336 samples), in order to characterise day/night and weekday/weekend DOC regimes along the plant. Samples were centrifuged before DOC determination or estimation. Four components were selected for describing wastewater composition and spectra. Reference spectra for these components were taken from the literature and proved to correctly explain the sample spectra. A two-step deconvolution method was developed, which avoided negative nitrate coefficients while keeping deconvolution error low. The calibration file for DOC estimation was determined by analysing DOC and acquiring UV spectra from 48 samples. DOC values were correlated to UV spectra by multiple linear regression. Determination coefficient and standard error were comparable to the values found in the literature. In raw or diluted samples with an absorbance between 2.0 and 2.5, DOC was probably underestimated by the method. This points to some nonlinearity for absorbances above 2.0, rather than the 2.5 limit suggested by the original method. DOC calculation through UV deconvolution allowed for the estimation of DOC regime along the WWTP. Time bands for higher and lower DOC concentrations were determined and characterised at each sampling point, for weekdays and the weekend. Except for the plant effluent, clear time bands were found. In effluent, DOC was always low, and very small oscillations were detected, due to DOC removal and intense mixing in the biological process. DOC profiles at each point are discussed in this paper. The modified deconvolution method has proved to be an accurate and efficient technique for estimating DOC of a large number of raw and diluted samples. PMID- 12753841 TI - Bulking in activated sludge plants treating paper mill wastewaters. AB - As part of a larger project to examine the causes of bulking in activated sludge plants treating wastewaters from paper mills, two laboratory-scale activated sludge plants were run in parallel. This was to evaluate the impact of variable environmental and operational parameters on the sludge ecology, and in particular, on the filamentous bacteria in the sludge. A survey of paper mills in the UK showed that there was a significant difference between the activated sludge plants at the paper mills using virgin fibre and those using recycled fibre. An examination of samples from activated sludge plants at the paper mills showed that there did not appear to be any significant correlation between filament abundance and the settlement characteristics as measured by stirred specific volume index (SSVI). The surface charge carried by the sludge particles was also measured and it was found that this parameter was better related to the SSVI. Similar results were found for samples from the laboratory plants. Seven different sets of operational conditions were applied to the laboratory-scale plants. These were aimed at simulating the conditions noted for the full-scale plants during the mill survey. The effects of low dissolved oxygen and low organic loading rates were examined. The effect of inorganic sulphur compounds and volatile fatty acids was studied. A change in raw material from virgin fibre to recycled fibre (corrugated paper) caused an immediate and continuous deterioration in settlement. Divalent cations, calcium and magnesium, were found to be successful in controlling settlement in the final clarifier. PMID- 12753842 TI - Role of hydraulic retention time and granular medium in microbial removal in tertiary treatment reed beds. AB - The main objective of this paper is to evaluate the role of hydraulic retention time (HRT) and granular medium in faecal coliform (FC) and somatic coliphage (SC) removal in tertiary reed beds. Experiments were carried out in a pilot plant with four parallel reed beds (horizontal subsurface flow constructed wetlands), each one containing a different type of granular medium. This pilot plant is located in a wastewater treatment plant in Montcada i Reixac, near Barcelona, in northeastern Spain. The microbial inactivation ratios obtained in the different beds are compared as a function of three selected HRTs. Secondary effluent from the wastewater treatment plant was used as the influent of the pilot system. The microbial inactivation ratio ranged between 0.1 and 2.7 log-units for FC and from 0.5 to 1.7 log-units for SC in beds with coarser granular material (5-25mm), while it ranged between 0.7 and 3.4 log-units for FC and from 0.9 to 2.6 log units for SC in the bed with finer material (2-13mm). HRT and granular medium are both key factors in microbial removal in the tertiary reed beds. The microbial inactivation ratio rises as the HRT increases until it reaches a saturation value (in general at an HRT of 3 days). The value of the microbial inactivation ratio at the saturation level depends on the granular medium contained in the bed. The specific surface area necessary to reach 2-3 log-units of FC and SC is approximately 3m(2)/person-equivalent. PMID- 12753843 TI - A reactive species model for chlorine decay and THM formation under rechlorination conditions. AB - Chlorine is typically used within drinking water distribution systems to maintain a disinfectant residual and minimize biological regrowth. Typical distribution system models describe the loss of disinfectant due to reactions within the water matrix as first order with respect to chlorine concentration, with the reactants in excess. Recent work, however, has investigated relatively simple dynamic models that include a second, hypothetical reactive species. This work extends these latter models to account for discontinuities associated with rechlorination events, such as those caused by booster chlorination and by mixing at distribution system junction nodes. Mathematical arguments show that the reactive species model will always represent chlorine decay better than, or as well as, a first-order model, under single dose or rechlorination conditions; this result is confirmed by experiments on five different natural waters, and is further shown that the reactive species model can be significantly better under some rechlorination conditions. Trihalomethane (THM) formation was also monitored, and results show that a linear relationship between total THM (TTHM) formation and chlorine demand is appropriate under both single dose and rechlorination conditions. This linear relationship was estimated using the modeled chlorine demand from a calibrated reactive species model, and using the measured chlorine demand, both of which adequately represented the TTHM formation. PMID- 12753844 TI - Estimating the precipitation potential in urine-collecting systems. AB - Precipitation in urine-separating toilets (NoMix toilets) and waterless urinals causes severe maintenance problems and can strongly reduce the content of soluble phosphate. In this study, we present a computer model for estimating the precipitation potential (PP) in urine-collecting systems. Calculating the PP enables to predict the composition and mass concentration of precipitates. We used our computer model for investigating how urea hydrolysis and dilution with flushing water affect precipitation. In a previous study, we found that microbial urea hydrolysis (ureolysis) triggers precipitation and that the amount of precipitates is limited by calcium and magnesium. With the present simulations, we could confirm these findings. We determined that only a small fraction of urea has to be hydrolysed for reaching 95% of the maximum PP. Since urease-positive bacteria are abundant in urine-collecting systems, strong precipitation is very likely. In further simulations, we determined that struvite (MgNH(4)PO(4).6H(2)O) and hydroxyapatite (HAP, Ca(10)(PO(4))(6)(OH)(2)) are the main precipitate compounds. If urine is highly diluted with tapwater, calcite (CaCO(3)) occurs as well. HAP is the only calcium phosphate mineral, although several others were supersaturated. Additionally, the simulations indicated that urine dilution diminishes the risk of blockages, since the mass concentration of precipitates decreases with the volume of flushing water. Rainwater flushing is more effective than flushing with tapwater. Moreover, flushing with tapwater leads to high phosphate fixation, because the total amount of calcium and magnesium ions increases, while the total amount of phosphate keeps constant. Finally, we compared simulation results with field measurements and found good agreement at low and very high urine dilution. PMID- 12753845 TI - Online titrimetric and off-gas analysis for examining nitrification processes in wastewater treatment. AB - The two steps of nitrification, namely the oxidation of ammonia to nitrite and nitrite to nitrate, often need to be considered separately in process studies. For a detailed examination, it is desirable to monitor the two-step sequence using online measurements. In this paper, the use of online titrimetric and off gas analysis (TOGA) methods for the examination of the process is presented. Using the known reaction stoichiometry, combination of the measured signals (rates of hydrogen ion production, oxygen uptake and carbon dioxide transfer) allows the determination of the three key process rates, namely the ammonia consumption rate, the nitrite accumulation rate and the nitrate production rate. Individual reaction rates determined with the TOGA sensor under a number of operation conditions are presented. The rates calculated directly from the measured signals are compared with those obtained from offline liquid sample analysis. Statistical analysis confirms that the results from the two approaches match well. This result could not have been guaranteed using alternative online methods. As a case study, the influences of pH and dissolved oxygen (DO) on nitrite accumulation are tested using the proposed method. It is shown that nitrite accumulation decreased with increasing DO and pH. Possible reasons for these observations are discussed. PMID- 12753846 TI - Survival tests with Chironomus riparius exposed to spiked sediments can profit from DEBtox model. AB - DEBtox model is a biologically based model used to analyse aquatic toxicity data (The analysis of aquatic toxicity data, VU University Press, Amsterdam, The Netherlands, 1996, 149pp.). To date, it has not been used to analyse sediment survival tests, mainly because this would require daily counting of survivors, which is difficult for benthic organisms. In the present study, we adapted survival toxicity tests with the midge Chironomus riparius to permit survival data analysis with DEBtox. To validate the adaptation proposed, we exposed organisms to two chemicals, copper and methiocarb. We exposed larvae of second, third and fourth instar to different concentrations of the tested compounds and two different diets to assess the influence of diet and instar on DEBtox parameter estimates. Daily counting of organisms did not affect survival and did not lead to much more effort when compared to usual tests. Moreover, the analysis profited much from use of the DEBtox model. It was possible, with only survival data to estimate and validate the kinetics of the compounds, to predict survival during the recovery of exposed organisms and to assess the influence of food availability on toxicity. Food availability did not influence methiocarb toxicity, but copper was more toxic to food-limited organisms when a threshold concentration was exceeded. Comparison of parameters estimate also helped in understanding the differences in toxicity responses between instars. For the two compounds we studied, the difference was entirely explained by differences in threshold values. PMID- 12753847 TI - Evidence for localized bacterial loading as the cause of chronic beach closings in a freshwater marina. AB - We conducted a comprehensive regional spatial assessment of bacterial water quality in order to determine the points of entry of fecal pollution into a swimming beach area on Lake Michigan that historically has had numerous water quality advisories for elevated levels of Escherichia coli (E. coli). Intensive, consecutive-day water samples were collected during dry and rainy conditions across multiple shoreline and offshore sites, and E. coli levels were enumerated from these samples. For both dry and rainy days, shoreline sites demonstrated significantly higher E. coli levels than offshore regions. We found that offshore (10-150m from shore) E. coli levels did not exceed 235CFU/100ml in more than 5% of the samples collected for 19 surveys (n=209). In contrast, samples taken at the beach area exceeded 235CFU/100ml in 66% of the samples collected for 43 shoreline surveys (n=675). Locally high E. coli levels coincided with bird presence and stormwater at the swimming beach located within the marina, and were unrelated to E. coli levels in connecting harbor waters. We conclude that beach water quality may be impacted by local, persistent contamination, which may confound routine beach monitoring and prevent the detection of regional pollution from other sources. PMID- 12753848 TI - Heavy metals extraction by microemulsions. AB - The objective of this study is the heavy metal extraction by microemulsion, using regional vegetable oils as surfactants. Firstly, the main parameters, which have influence in the microemulsion region, such as: nature of cosurfactant, influence of cosurfactant (C)/surfactant (S) ratio and salinity were studied, with the objective of choosing the best extraction system. The extraction/reextraction process by microemulsion consists of two stages. In the first one, the heavy metal ion present in the aqueous phase is extracted by the microemulsion. In a second step, the reextraction process occurs: the microemulsion phase, rich in metal, is acidified and the metal is recovered in a new aqueous phase, with higher concentration. The used system had the following parameters: surfactant saponified coconut oil; cosurfactant-n-butanol; oil phase-kerosene; C/S ratio=4; salinity-2% (NaCl); temperature of 27+/-1 degrees C; water phase-aqueous solution that varied according to the heavy metal in study (Cr, Cu, Fe, Mn, Ni and Pb). A methodology of experimental planning was used (Scheffe Net) to study the behavior of the extraction in a chosen domain. The extraction was accomplished in one step and yielded extraction percentage higher than 98% for all metals. In the reextraction HCl-8M was used as reextraction agent and the influence of the pH and time were verified. This work showed the great efficiency of the microemulsion, indicating that it is possible to extract selectively the heavy metals from the aqueous phase. PMID- 12753849 TI - Algal growth inhibition test: does shading of coloured substances really matter? AB - Coloured substances are often assumed to behave differently in the algal growth inhibition test due to shading of the algae. We have investigated EU notifications of new substances and found no significant differences in algal test results between colours and non-colours. A modified method to differentiate between shading and toxicity is not satisfactory, because the method used to determine the shading effect is incorrect. Due to the shaking of the test flasks in the algal test intermittent light conditions are achieved which lead to a higher growth rate than expected from proposed tests with constant reduced light conditions. This will result in overestimation of the shading effect and thus to underestimation of chemical toxicity. Furthermore, we showed in experiments with the reference substance potassium dichromate that reduced light can reduce the measurable toxicity of chemicals. Comparing our results with approaches proposed in the literature, a simple approach to reduce effects of undesired shading when testing coloured substances is to achieve light saturation for the algae by increasing illumination at least to the upper limit given in test guidelines (120 microEs(-1)m(-2)) and, optionally, to reduce the light path by choosing smaller test volumes. PMID- 12753850 TI - Application of calorimetric measurements for biokinetic characterisation of nitrifying population in activated sludge. AB - A preliminary investigation is described on the application of calorimetry as a sensitive technique to evaluate nitrifying activity in activated sludge. Calorimetric profiles (thermograms) related to heat dissipation due to biological nitrification reactions (ammonia or nitrite consumption) have been interpreted. Correlations between calorimetric data and the main process variables, i.e. ammonia and nitrite concentration and oxygen uptake, have been verified, and confirm the potential of calorimetry to investigate, monitor and control even weakly exothermic biological processes like autotrophic nitrification. Heat yields (Y(Q/i)) for ammonia, nitrite, and oxygen, defined as the heat released per unit amount of converted reactant, have been separately evaluated. Moreover, calorimetric experiments on activated sludge from a full-scale nitrogen removal wastewater treatment plant have been carried out and kinetic parameters for both ammonia and nitrite oxidising bacteria have been estimated. PMID- 12753851 TI - Study on the use of NADH fluorescence measurements for monitoring wastewater treatment systems. AB - Fluorescence measurement of intracellular nicotinamide adenine dinucleotide (NADH) provides information about the physiological response of microbes towards changing conditions in their environment and has been suggested to be useful for the control of wastewater treatment plants. In this study, the practical usefulness of such measurements was evaluated from batch experiments with a commercially available NADH sensor in a bench scale reactor. The sensor was linear in the NADH concentration, robust, almost maintenance free, and hardly sensitive to floc size distribution. Measured fluorescence intensity proved to depend strongly on the concentration of active heterotrophic biomass. The NADH level was supposed to be dependent on the ratio of electron donor/electron acceptor availability inside the cells; however, neither acetate nor ammonium addition was reflected by the measurement signal.A jump wise NADH signal change was observed at complete oxygen or nitrate depletion as also reflected by bends in the redox curve. In the near zero concentration ranges of oxygen and nitrate (0.1-0.5mg/l) the signal changes only slightly in the opposite direction to the redox trend. PMID- 12753852 TI - Floc morphology and size distributions of cohesive sediment in steady-state flow. AB - Fractal dimensions of particle populations of cohesive sediment were examined during deposition experiments in an annular flume at four conditions of steady state flow (0.058, 0.123, 0.212 and 0.323Pa). Light microscopy and an image analysis system were used to determine area, longest axis and perimeter of suspended solids. Four fractal dimensions (D, D(1), D(2), D(k)) were calculated from the slopes of regression lines of the relevant variables on double log plots. The fractal dimension D, which relates the projected area (A) to the perimeter (P) of the particle (P proportional, variant A(D/2)), increased from 1.25+/-0.005 at a shear stress of 0.058Pa to a maximum of 1.36+/-0.003 at 0.121Pa then decreased to 1.34+/-0.001 at 0.323Pa. The change in D indicated that particle boundaries became more convoluted and the shape of larger particles was more irregular at higher levels of shear stress. At the highest shear stress, the observed decrease in D resulted from floc breakage due to increased particle collisions. The fractal dimension D(1), which relates the longest axis (l) to the perimeter of the particle (P proportional to l(D1)), increased from 1.00+/-0.006 at a shear stress of 0.058Pa to a maximum of 1.25+/-0.003 at 0.325Pa. The fractal dimension D(2), which relates the longest axis with the projected area of the particle (A proportional to l(D(2)), increased from 1.35+/-0.014 at a shear stress of 0.058Pa to a maximum of 1.81+/-0.005 at 0.323Pa. The observed increases in D(1) and D(2) indicate that particles became more elongated with increasing shear stress. Values of the fractal dimension D(k), resulting from the Korcak's empirical law for particle population, decreased from 3.68+/-0.002 at a shear stress of 0.058Pa to 1.33+/-0.001 at 0.323Pa and indicate that the particle size distribution changed from a population of similar sized particles at low shear to larger flocculated particles at higher levels of shear. The results show that small particle clusters (micro-flocs) are the formational units of larger flocs in the water column and the stability of larger flocs is a function of the shear stress at steady state. PMID- 12753853 TI - Combined advanced oxidation and biodegradation of industrial effluents from the production of stilbene-based fluorescent whitening agents. AB - Three different industrial wastewaters from the production of stilbene-based fluorescent whitening agents were investigated with regard to the applicability of advanced oxidation processes combined with biodegradation. Oxidation processes included the application of ozone, hydrogen peroxide, UV-radiation and Fenton's reagent (Fe(2+)/H(2)O(2)). Characterization of the combined chemical-biological treatment was done by sum parameters and HPLC analysis. In addition, toxicity was determined using the luminescence inhibition test. Results showed that processes producing OH-radicals without the need of UV-irradiation proved to be suited for the oxidation of all three wastewaters. H(2)O(2)/UV processes were ineffective due to the high inner filter effect of the effluents. Comparing the combined oxidative-biological process with biological treatment, the applied pre-oxidation steps did not always lead to a significant improvement of the biological degradation. In one case, an inverted treatment starting with biodegradation followed by oxidation turned out to be the preferable procedure. After oxidation with ozone or ozone combined with UV-irradiation, an increase in toxicity was partly observed indicating the formation of toxic intermediate products. In some cases samples had to be diluted before the biodegradation step to achieve a better biodegradability. PMID- 12753854 TI - High performance degradation of azo dye Acid Orange 7 and sulfanilic acid in a laboratory scale reactor after seeding with cultured bacterial strains. AB - Bacterial strains 1CX and SAD4i--previously isolated from the mixed liquor of a municipal sewage treatment plant--are capable of degrading the azo dye Acid Orange 7 (AO7) and sulfanilic acid, respectively. A rotating drum bioreactor (RDBR), operating under continuous flow and nutrient conditions designed to simulate the effluent from a dye manufacturing plant, was seeded with strains 1CX and SAD4i, forming a biofilm capable of degrading AO7 and sulfanilic acid. In addition, an RDBR containing a pre-existing biofilm capable of degrading AO7, but not sulfanilic acid, was seeded with strain SAD4i alone. Strain SAD4i was incorporated into the existing biofilm and degraded the sulfanilic acid resulting from the degradation of AO7 by indigenous members of the biofilm. The ability to seed a bioreactor with bacterial strains capable of degrading azo dyes, and resulting by-products, in a mixed microbial community suggests that this process could have commercial applications. PMID- 12753855 TI - Effect of periodic feeding in sequencing batch reactor on substrate uptake and storage rates by a pure culture of Amaricoccus kaplicensis. AB - A pure culture of Amaricoccus kaplicensis was aerobically cultured at a long culture residence time (Theta(C)>12d), under periodic acetate feeding in a sequencing batch reactor (SBR). The cycle length and, correspondingly, the volumetric organic load rate (vOLR) were varied in the range 4-24h and 0.76 0.12gCODl(-1)d(-1), respectively. The transient response of the microorganism to the acetate spike was investigated throughout batch tests, as a function of SBR cycle length and vOLR. In all tested conditions, a rapid transient response was observed, mainly due to acetate storage in the form of polyhydroxybutyrate, since growth (production of active biomass) played a minor role. Apart from this general trend, the maximum rates under transient conditions increased as the cycle length increased from 4 to 24h. In the SBR, the longest cycle also caused a decrease in floc size and settleability as well as an increase in the observed yield. The observed effect of SBR operating conditions on the physiological state of cells and their related transient response may have great significance on the performance of full scale activated sludge processes. PMID- 12753856 TI - Development of a Ct equation for the inactivation of Cryptosporidium oocysts with chlorine dioxide. AB - Cryptosporidium parvum, a protozoan parasite, has been implicated in a number of waterborne disease outbreaks and is difficult to inactivate using free chlorine. It appears, however, to be inactivated more easily by other oxidants such as chlorine dioxide or ozone. A major element of the EPA (US EPA) strategy for controlling C. parvum (oocysts) in drinking water is the possible use of Ct (concentration of disinfectant in mg/L times time in minutes) values. To support this strategy a Ct equation, based on first-order kinetics, is proposed to provide guidance to drinking water utilities for the application of chlorine dioxide for controlling C. parvum oocysts. The equation is based on standard statistical techniques using available bench scale data. It can predict mean inactivation levels as well as a statistically conservative upper bound Ct value. This upper bound could be used to insure an appropriate safety factor for the protection of public health. PMID- 12753857 TI - Biofilm in the sediment phase of a sanitary gravity sewer. AB - Microbial activity of the sediment phase in a 1.5-km-long concrete sewer section with a cement pipe in a 540-mm diameter was investigated in this paper. SEM examinations and elementary composition analyses of the sediment samples have identified the presence of a biofilm layer at the sediment surface. Bacterial counting results with a DNA-staining technique have revealed that the amount of bacteria in this layer was 2.1 x 10(11)cellg(-1) dry wt, which is close to that of activated sludge. ATP content in the sewer biofilm was found relatively high, demonstrating that the sewer biofilm is active. Throughout the entire 1.5-km sewer section, the biofilm activity was maintained at almost the same level. Lab scale sediment oxygen uptake flux (SOUF) tests showed that the shear flow velocity above the sediment phase linearly increases the SOUF, which of the potential value was determined to be 32gO(2)m(-2) day(-1) at an estimated shear flow velocity of 0.055ms(-1) at 25 degrees C in the sewer line, provided that the mean flow velocity was 1.5ms(-1), and the mean water depth was 220mm. Such a high SOUF value further endorsed the existence of the active sewer biofilm. PMID- 12753858 TI - Using filtrate of waste biosolids to effectively produce bio-hydrogen by anaerobic fermentation. AB - Waste biosolids collected from sewage works is a biomass containing a vast amount of polysaccharides and proteins, and thus is considered a potential substrate for producing hydrogen using anaerobic fermentation. This work demonstrated, contrary to the common assumption, that the solids phase in waste activated biosolids presents extra nutrients for anaerobes; it in fact prohibits effective bio hydrogen production. Using filtrate after removal of solids from biosolids produces more hydrogen than using the whole biosolids, with the former reaching a level an order of magnitude higher than the literature results. PMID- 12753859 TI - Comment on "Determination of areal sedimentation rates in rivers by using plate sediment trap measurements and flow velocity-settling flux relationship" by Hans Peter Kozerski. PMID- 12753861 TI - Systemic application of growth hormone enhances the early healing phase of osteochondral defects--a preliminary study in micropigs. AB - Healing of osteochondral defects following trauma remains a significant clinical problem, often leading to osteoarthritis. Growth hormone (GH) has been shown to accelerate formation of bone and cartilage tissue in the growth plates and in cell cultures. To investigate the influence of systemically administered recombinant porcine growth hormone (r-pGH) on the healing of osteochondral defects we performed a histomorphometrical analysis of full-thickness cartilage defects in the femoral condyle of micropigs. Forty-eight mature female Yucatan micropigs were divided into two groups, one receiving a daily injection of r-pGH (100 microg/kg), the other receiving sodium chloride as placebo. A circular 6-mm diameter full-thickness defect of the cartilage was created, extending 1.5 mm into the subchondral bone. The animals were sacrificed after 4 (n = 24) and 6 (n = 24) weeks. The von-Kossa stain was used to visualise the calcified structures; cartilage and the fibrous tissue were marked with a combined Safranin-O/light green stain. The defect filling and the percentage of bone, cartilage, and fibrous tissue into the defect were evaluated using an image analysis system. Furthermore, histological grading was performed using the modified Wakitani score. After 4 weeks no differences were observed between both groups. The defect filling after 6 weeks with newly formed bone was significantly higher in the r pGH-treated group. The formation of cartilage and fibrous tissue showed a trend towards better healing in the GH-treated group; however, there was no significant difference. In the r-pGH-treated group, the percentage of total defect filling was significantly higher. The evaluation of the vascularity showed a significantly lower number of vessels in the GH-treated group after 6 weeks. Histomorphological grading revealed a significantly lower total Wakitani score in the GH-treated group, which represents a better healing result compared to the controls. The results of the present study suggest that circulating r-pGH or one of its mediators may accelerate osteochondral defect healing by stimulating the formation of osseous and chondral tissue. The analysis of the vascularity leads to the assumption of an advanced maturation of the osteochondral defects under the influence of GH. PMID- 12753862 TI - The components of excess mortality after hip fracture. AB - A high excess mortality is well described after hip fracture. Deaths are in part related to comorbidity and in part due directly or indirectly to the hip fracture event itself (causally related deaths). The aim of this study was to examine the quantum and pattern of mortality following hip fracture. We studied 160,000 hip fractures in men and women aged 50 years or more, in 28.8 million person-years from the patient register of Sweden, using Poisson models applied to hip fracture patients and the general population. At all ages the risk of death was markedly increased compared with population values immediately after the event. Mortality subsequently decreased over a period of 6 months, but thereafter remained higher than that of the general population. The latter function was assumed to account for deaths related to comorbidity and the residuum assumed to be due to the hip fracture. Causally related deaths comprised 17-32% of all deaths associated with hip fracture (depending on age) and accounted for more than 1.5% of all deaths in the population aged 50 years or more. Hip fracture was a more common cause for mortality than pancreatic or stomach cancer. Thus, interventions that decreased hip fracture rate by, say, 50% would avoid 0.75% or more of all deaths. PMID- 12753863 TI - Identification of genes regulated during osteoblastic differentiation by genome wide expression analysis of mouse calvaria primary osteoblasts in vitro. AB - Although several independent studies of gene expression patterns during osteoblast differentiation in cultures from calvaria and other in vitro models have been reported, only a small portion of the mRNAs expressed in osteoblasts have been characterized. We have previously analyzed the behavior of several known markers in osteoblasts, using Affymetrix GeneChip murine probe arrays (27,000 genes). In the present study we report larger groups of transcripts displaying significant expression modulation during the culture of osteoblasts isolated from mice calvaria. The expression profiles of 601 such regulated genes, classified in distinct functional families, are presented and analyzed here. Although some of these genes have previously been shown to play important roles in bone biology, the large majority of them have never been demonstrated to be regulated during osteoblast differentiation. Despite the fact that the precise involvement of these genes in osteoblast differentiation and function needs to be evaluated, the data presented herein will aid in the identification of genes that play a significant role in osteoblasts. This will provide a better understanding of the regulation of osteoblast differentiation and maturation. PMID- 12753864 TI - Expression of inducible cAMP early repressor is coupled to the cAMP-protein kinase A signaling pathway in osteoblasts. AB - We previously showed that parathyroid hormone (PTH) induces inducible cAMP early repressor (ICER) in osteoblastic cells and mouse calvariae. PTH signaling in osteoblastic cells is transduced by PTH receptor 1, which is coupled to cAMP protein kinase A (PKA), protein kinase C (PKC), and calcium signaling pathways. In the present study, we examined the role of these pathways in mediating PTH induced ICER mRNA and protein expression in osteoblastic MC3T3-E1 cells. Using RT PCR, we found that PTH(1-34), forskolin (FSK), and 8-bromo-cAMP (8Br-cAMP) induced ICER expression, while phorbol myristate acetate (PMA), ionomycin, and PTH(3-34) did not. Similar results were found for the induction of ICER protein. PKA inhibition by H89 markedly reduced PTH- and FSK-induced ICER expression, while PKC depletion by PMA had little effect. We also tested ICER induction by other osteotropic signaling agonists. Other cAMP-PKA pathway activators, such as PTH-related protein (PTHrP), induced ICER expression, while agents that signal through other pathways did not. PTHrP maximally induced ICER mRNA at 2-4 h, which then returned to baseline by 10 h. Finally, PTH, FSK, and PTHrP induced ICER in cultured mouse calvariae and osteoblastic ROS 17/2.8, UMR-106, and Pyla cells. We conclude that ICER expression in osteoblasts requires activation of the cAMP-PKA signaling pathway. PMID- 12753865 TI - Vascular endothelial growth factor is expressed along with its receptors during the healing process of bone and bone marrow after drill-hole injury in rats. AB - In this study, we investigated the expression of vascular endothelial growth factor (VEGF) mRNA along with its receptors in the healing process following rat femoral drill-hole injury. The cellular events involved in the differential expression of VEGF were studied by reverse transcription-polymerase chain reaction, immunocytochemistry, and in situ hybridization. Abundant alkaline phosphatase-positive osteoprogenitor cells were present in the bone marrow cavity surrounding the wound region at day 3. Some of the cells were immunoreactive for Flk-1, a marker of angioblasts. At day 5, osteoblasts expressing osteocalcin mRNA actively participated in bone formation. After day 11, medullary bone gradually decreased and hematopoietic cells covered the wound region. The expressions of the VEGF splice variants VEGF120 and VEGF164 were detected at days 1 and 3, and VEGF188 mRNA began to appear from day 5. The expressions of the three VEGF splice variants gradually decreased after day 11. VEGF immunoreactivity and mRNA expression were strongly detected in angioblasts, osteoprogenitor cells, and osteoblasts between days 3 and 7, but gradually decreased after day 11. Immunoreactivity for Flt-1 was also detected in endothelial cells, osteoprogenitor cells, and osteoblasts between days 3 and 7. However, immunoreactivity for Flk-1 was not detected on osteoblasts but rather on endothelial cells. These findings indicate that the differential expression of VEGF splicing isoforms along with its receptors may play an important role in the healing process after rat femoral drill-hole injury. PMID- 12753867 TI - Anabolic action of parathyroid hormone on cortical and cancellous bone differs between axial and appendicular skeletal sites in mice. AB - The mouse is being increasingly used to study the anabolic action of parathyroid hormone (PTH) on the skeleton. The efficacy of intermittent PTH treatment on bone varies widely among tested strains of mice with differences in peak bone mass and structure. We have therefore examined the responses of skeletal sites with high or low cancellous bone mass to PTH treatment in a single strain with genetically low bone mass. Mature C57BL/6 mice were ovariectomized (ovx) or sham operated and, after 4 weeks, treated with PTH(1-34) (40 microg/kg/day, 5 days/week sc) or vehicle for 3 or 7 weeks. Two doses of fluorescent labels were given to the animals 9 and 3 days before euthanasia. Histomorphometry was performed on sections of the proximal tibia, tibial diaphysis, and vertebral body. The results indicate that 4 to 11 weeks of ovx induced a approximately 44% loss of cancellous bone in the proximal tibia and a approximately 25% loss of cancellous bone in the vertebra with impaired trabecular architecture and high bone turnover. In the intact animals, PTH increased cancellous bone volume to a greater extent in the vertebral body than in the proximal tibia, a site with lower cancellous bone volume at the outset. In the ovx mice, PTH increased cancellous bone volume to a greater extent in the vertebral body, a site displaying moderate cancellous bone loss, than in the proximal tibia, a site with severe cancellous bone loss. Conversely, the treatment added a little cortical bone to the tibia, a highly loaded site, but did not significantly increase cortical width of the vertebral body, a less loaded site. We conclude that, for intermittent PTH treatment to be maximally effective, there must be an adequate number of trabeculae present at the beginning of treatment, regardless of estrogen status. Our results also support an interaction between PTH anabolic action and mechanical loading. PMID- 12753866 TI - Transplantation of skin fibroblasts expressing BMP-2 promotes bone repair more effectively than those expressing Runx2. AB - We investigated the osteogenic potential of skin fibroblasts that overexpressed BMP-2 or Runx2 by using adenoviral vectors. In in vitro experiments, skin fibroblasts infected with adenovirus vector encoding BMP-2 (AdBMP-2) released substantial levels of BMP-2 proteins into culture media, and those infected with adenovirus vector encoding Runx2 (AdRunx2) produced its protein. Transduction of BMP-2 or Runx2, respectively, increased alkaline phosphatase (ALP) activity and induced expression of mRNAs of ALP, osteocalcin, and osterix in skin fibroblasts. In in vivo experiments, we investigated the bone induction activity by transplantation of a complex composed of carrier [poly-D,L-lactic-co-glycolic acid/gelatin sponge (PGS)] and skin fibroblasts (PGS/SF complex). Transplantation of PGS/SF complexes composed of skin fibroblasts transduced with AdBMP-2-induced ectopic bone formation when transplanted into the subfascia of back muscle, unlike those infected with AdRunx2. Transplantation of PGS/SF complexes composed of skin fibroblasts transduced with AdBMP-2 into craniotomy defects induced bone formation from 2 weeks after transplantation, and almost all PGS was replaced by newly synthesized bone at 6 weeks. To investigate the fate of the transplanted cells, we transplanted skin fibroblasts isolated from green fluorescence protein transgenic mice into craniotomy defects. Transplantation of these skin fibroblasts transfected with AdBMP-2 generated green fluorescence protein positive osteoblasts and osteocytes, indicating that the transplanted skin fibroblasts differentiated into osteoblastic lineage cells during bone repair. In contrast, transplantation of PGS/SF complexes composed of skin fibroblasts transduced with AdRunx2 induced a few ALP-positive cells at 1 week after transplantation, but their number decreased depending on time after transplantation. In addition, transplantation of these complexes was insufficient to induce bone repair. Taken together, our results suggest that skin fibroblasts expressing BMP-2 are more suitable for cell-mediated therapy of bone repair than those expressing Runx2. PMID- 12753868 TI - Gentamicin coating of metallic implants reduces implant-related osteomyelitis in rats. AB - Antibiotic prophylaxis is a routine procedure in orthopedic surgery. Various local antibiotic delivery techniques are used to reduce bone- and soft tissue related infection. The objective of this study was to evaluate the efficacy of a new biodegradable, gentamicin-loaded poly(D,L-lactide) (PDLLA) coating of orthopedic devices in preventing implant-related osteomyelitis. The medullary cavities of tibiae in 30 Sprague Dawley rats were contaminated with Staphylococcus aureus (10(3) colony forming units). Simultaneously titanium Kirschner wires, uncoated (group II), coated with PDLLA (group III), or coated with PDLLA + 10% gentamicin (group IV), were implanted. Ten animals that received phosphate-buffered saline and uncoated Kirschner wires served as controls (group I). Follow-up was 6 weeks. In weekly intervals X-rays of the tibiae were performed, blood counts were taken, and body temperature and weight were determined. After sacrifice infection was evaluated by histological and microbiological analysis. All animals of groups II and III developed microbiological, histological, and radiological signs of infection, including osseous destruction and soft tissue swelling. All animals of the control group remained sterile. Cultures of implants of group IV showed significantly reduced bacterial growth compared to cultures of groups II and III, and three implants of group IV remained sterile. Further radiological and histological signs of infection were significantly reduced in the gentamicin-coated group compared to groups II and III. No significant differences in body weight, body temperature, and blood parameters between all groups were observed. Local application of antibiotic-coated orthopedic devices containing PDLLA and 10% gentamicin significantly reduced implant-related infection in this animal model. PMID- 12753869 TI - Co-dependence of calcium and phosphorus for growth and bone development under conditions of varying deficiency. AB - The purpose of this study was to quantify the effect of variation in calcium intake, with and without supplemental phosphorus, on growth and bone development in growing animals under conditions of varying calcium and phosphorus deficiency. Nine groups of weanling male rats were fed a diet nutritionally complete, except for calcium and phosphorus, for 28 days. This diet provided nine levels of varying calcium and phosphorus repletion, using either calcium carbonate, dicalcium phosphate, or tricalcium phosphate. Body weights and diet consumption were measured throughout the test period. At term, the femurs from each animal were weighed, measured for tensile strength, bone mineral content (BMC), and bone density, and analyzed for ash, calcium, phosphorus, and histology. As expected, at equivalent levels of calcium supplementation, the two phosphorus-containing salts promoted significantly greater improvement in all the bone variables measured, as well as greater body weight gain and diet and calcium utilization, compared to animals supplemented with calcium only. Histomorphometric analysis confirmed the results of the mineral analysis and showed the structural impact of the inadequate mineral intake. The mean values for ash weight, BMC, and tensile strength in the nine diet groups were well fit (R(2) values ranging from 0.93 to 0.99) by multivariate models incorporating only the diet content values for calcium and phosphorus. In these models, the value for the phosphorus coefficient was three to sixfold larger than that for the calcium term, indicating a substantially greater effect of varying phosphorus intake than of varying calcium. These results demonstrate both the co-dependence of calcium and phosphorus in bone development and the importance of providing both minerals to support soft tissue and bone growth. PMID- 12753870 TI - Subchondral tibial bone mineral density predicts future joint space narrowing at the medial femoro-tibial compartment in patients with knee osteoarthritis. AB - Preliminary studies have shown that dual-energy X-ray absorptiometry (DXA) produces images of sufficient quality for a precise and accurate measurement at density of the subchondral bone. The objective of this study was to investigate the relationship between baseline subchondral tibial bone mineral density (BMD) and joint space narrowing observed after 1 year at the medial femoro-tibial compartment of the knee joint. Fifty-six consecutive patients, from both genders, with knee osteoarthritis diagnosed according to the American College of Rheumatology criteria, were included in the study. Radiographic posteroanterior views were taken, at baseline and after 1 year of follow-up. Minimum joint space width (JSW) measurement, at the medial femoro-tibial joint, was performed with a 0.1-mm graduated magnifying lens. Baseline BMD of the subchondral tibial bone was assessed by DXA. The mean +/- SD age of the patients was 65.3 +/- 8.7 years, with a body mass index of 28.0 +/- 4.9 kg/m(2). The minimum JSW was 3.5 +/- 1.5 mm and the mean BMD of the subchondral bone was 0.848 +/- 0.173 g/cm(2). There was a significant negative correlation between subchondral BMD and 1-year changes in minimum JSW (r = -0.43, p = 0.02). When performing a multiple regression analysis with age, sex, body mass index, and minimum JSW at baseline as concomitant variables, BMD of the subchondral bone as well as JSW at baseline were independent predictors of 1-year changes in JSW (p = 0.02 and p = 0.005, respectively). Patients in the lowest quartile of baseline BMD (<0.73 g/cm(2)) experienced less joint space narrowing than those in the highest BMD quartile (>0.96 g/cm(2)) (+0.61 +/- 0.69 mm versus -0.13 +/- 0.27 mm; p = 0.03). Assessment of BMD of the subchondral tibial bone is significantly correlated with future joint space narrowing and could be used as a predictor of knee osteoarthritis progression. PMID- 12753871 TI - Peak spine and femoral neck bone mass in young women. AB - Achievement of higher peak bone mass early in life may play a critical role against postmenopausal bone loss. Bone mineral density (BMD) of the spine, femoral neck, greater trochanter, Ward's triangle, and spine bone mineral content (BMC) and bone surface area (BSA) were assessed by dual energy x-ray absorptiometry in 300 healthy females (age 6-32 years). Bone measurements were described by using nonlinear models with age, weight, height, or dietary calcium intake as the explanatory variables. At the spine, femoral neck, greater trochanter, and Ward's triangle, the highest BMD level was observed at 23.0 +/- 1.4, 18.5 +/- 1.6, 14.2 +/- 2.0, and 15.8 +/- 2.1 years, respectively. The age of attaining peak spine BMC and BSA cannot be estimated, as significant increases in these two measures were observed through this age group. Age, weight, and height were all significant predictors of all these bone measurements. Weight was a stronger predictor than age for all sites. Dietary calcium intake was not a significant predictor for any of these bone measurements. We conclude that age of attaining peak bone mass at the hip is younger than at the spine, and BMC and BSA at the spine continue to increase through the early thirties in females. PMID- 12753872 TI - Quantitative trait loci for periosteal circumference (PC): identification of single loci and epistatic effects in F2 MRL/SJL mice. AB - To test the hypothesis that periosteal circumference (PC), which is associated with bone size through cross-sectional moment of inertia (CMI), has heritable components, we performed a linkage analysis using 633 MRL/SJL F(2) mice that have 14% difference in mean PC. PC was determined in femurs by use of peripheral quantitative computerized tomography (pQCT). The genome-wide scan identified nine QTL for PC adjusted by body weight on chromosomes 1 (2 QTL), 2 (2 QTL), 8, 11, 15, 17, and X, which accounted for 38.6% of phenotype variance. QTL on chromosomes 1 (D1Mit33), 8 (D8Mit125), 15 (D15Mit 62), 17 (D17Mit176), and X (DXMit208) were unique for PC adjusted by body weight and femur length, while the remaining PC QTL were shared with body weight but not femur length. Four epistatic interactions were identified which accounted for 37.6% of phenotype variance. There was also evidence of pleiotropic effects on chromosome 11 among four size phenotypes (PC, body length, body weight, bone mineral density, and muscle size), which may represent a common genetic mechanism that may regulate bone size and body size. PMID- 12753873 TI - Effects of gender, anthropometric variables, and aging on the evolution of hip strength in men and women aged over 65. AB - Although gender differences in fall rates may partly explain the higher prevalence of fractures in elderly women than men, male bones may also be intrinsically stronger or suffer less structural degradation with age than those of women. We used hip structural analysis (HSA) to study gender differences in hip geometry and bone mineral density (BMD) as they evolved over time in elderly white men and women with the aim of identifying candidate biological pathways leading to heightened risk of hip fracture. We recruited 443 women and 439 men aged 67-79 years from a diet and cancer prospective population-based cohort study to a study of hip bone loss. Hip BMD was measured on two occasions 2-5 years apart by dual-energy X-ray absorptiometry and HSA software used to derive BMD and structural parameters at the narrow neck (NN), the intertrochanter (IT), and the shaft (S) regions. Structural indices calculated in each region were cross sectional area (CSA)-amount of bone surface area in the cross section after excluding soft tissue space; section modulus (Z)-an index of bending resistance, subperiosteal width, endocortical width, cortical thickness; and cortical buckling ratio (CBR)-a measure of cortical instability. Compared to men, women had lower values of BMD, CSA, Z, subperiosteal width, endocortical width, and cortical thickness in all regions, except S endocortical width, after adjusting for weight, height, and age (P < 0.0001). CBR was higher in women than in men (P < 0.0001) in all regions. Longitudinal analysis of rates of change revealed faster rates of BMD decline in women than in men at the Hologic total hip, Hologic femoral neck, and IT regions (P < 0.029). Women had faster rates of subperiosteal and endosteal expansion than men at the NN (P < 0.011) and IT (P < 0.049) and faster increase in Z at the NN (P = 0.029). At the IT region, cortical thinning was faster in women than in men (P = 0.037) and CBR increased at a faster rate in women (P = 0.011). In conclusion, Z is lower in women than in men and expansion of the proximal femur occurs in both sexes, being faster in women than in men. Z does not decline at the same rate as BMD, implying that part of the effect of aging on BMD is due to expansion of the bony envelope without loss of bone mineral content. Faster expansion in the female femoral neck may in turn lead to greater fragility if wider diameter and thinner cortices become locally unstable. PMID- 12753874 TI - Performance of quantitative ultrasound in the discrimination of prevalent osteoporotic fractures in a bone metabolic unit. AB - There is a growing interest in ultrasound evaluation of bone status as an alternative to the measurement with dual X-ray absorptiometry (DXA), due to its low cost, portability, and nonionizing radiation. The aim of our study was to investigate the relation among DXA, QUS, clinical, anthropometric, and lifestyle factors, and to determine QUS cutoff values in order to discriminate fractures in patients referred to the Bone Metabolic Unit at an Endocrinology Service. We studied 300 patients (281 females and 19 males; age 58 +/- 11 years) referred for evaluation of osteoporosis. In all cases we determined basic anthropometric parameters, a clinical history including previous osteoporotic fractures and risk factors for osteoporosis, and QUS parameters in calcaneus (Hologic Sahara), and BMD in lumbar spine (LS) and femoral neck (FN), by DXA (Hologic QDR 1000). Using the WHO densitometric criteria, 37, 46.7, and 16.3% of our population were osteoporotic, osteopenic, and normal, respectively. A QUI T-score or=4 (95% [39/41]) than in those with undetectable antibodies (13% [9/70]; P <0.0001), a titer of 1 (18% [4/22]; P <0.0001), or a titer of 2 (33% [2/6]; P = 0.001). All 19 samples with a positive platelet aggregation test had anti heparin/PF4 antibody titers of at least 4, including 15 samples with titers >or=32. Thromboembolic complications in heparin-treated patients were significantly more prevalent in patients with titers >or=4 (63% [26/41]) than in those with undetectable antibodies (8% [6/79]; P <0.0001) or a titer of 1 (9% [2/22]; P <0.0001). Of the 11 patients with a titer of 1 who were maintained on heparin, none developed worse thrombocytopenia or thromboembolic complications. CONCLUSION: Anti-heparin/PF4 antibody titers, which can be measured rapidly and reproducibly using a particle gel immunoassay, can be used as a confirmatory test to complement a clinical likelihood score among patients with suspected heparin induced thrombocytopenia. PMID- 12753877 TI - Tramadol and acetaminophen combination tablets in the treatment of fibromyalgia pain: a double-blind, randomized, placebo-controlled study. AB - PURPOSE: To evaluate the efficacy and safety of a combination analgesic tablet (37.5 mg tramadol/325 mg acetaminophen) for the treatment of fibromyalgia pain. METHODS: This 91-day, multicenter, double-blind, randomized, placebo-controlled study compared tramadol/acetaminophen combination tablets with placebo. The primary outcome variable was cumulative time to discontinuation (Kaplan-Meier analysis). Secondary measures at the end of the study included pain, pain relief, total tender points, myalgia, health status, and Fibromyalgia Impact Questionnaire scores. RESULTS: Of the 315 subjects who were enrolled in the study, 313 (294 women [94%], mean [+/- SD] age, 50 +/- 10 years) completed at least one postrandomization efficacy assessment (tramadol/acetaminophen: n = 156; placebo: n = 157). Discontinuation of treatment for any reason was less common in those treated with tramadol/acetaminophen compared with placebo (48% vs. 62%, P = 0.004). Tramadol/acetaminophen-treated subjects also had significantly less pain at the end of the study (53 +/- 32 vs. 65 +/- 29 on a visual analog scale of 0 to 100, P <0.001), and better pain relief (1.7 +/- 1.4 vs. 0.8 +/- 1.3 on a scale of -1 to 4, P <0.001) and Fibromyalgia Impact Questionnaire scores (P = 0.008). Indexes of physical functioning, role-physical, body pain, health transition, and physical component summary all improved significantly in the tramadol/acetaminophen-treated subjects. Discontinuation due to adverse events occurred in 19% (n = 29) of tramadol/acetaminophen-treated subjects and 12% (n = 18) of placebo-treated subjects (P = 0.09). The mean dose of tramadol/acetaminophen was 4.0 +/- 1.8 tablets per day. CONCLUSION: A tramadol/acetaminophen combination tablet was effective for the treatment of fibromyalgia pain without any serious adverse effects. PMID- 12753878 TI - Potential effect of cyclooxygenase-2-specific inhibitors on the prevention of colorectal cancer: a cost-effectiveness analysis. AB - PURPOSE: To estimate the potential cost-effectiveness of colorectal cancer chemoprevention with cyclooxygenase-2-specific inhibitors (COX-2 inhibitors). METHODS: Using a decision analytic Markov model, we estimated the discounted cost per life-year saved for three strategies: a COX-2 inhibitor alone; as an adjunct to colonoscopy every 10 years in persons at average risk of colorectal cancer; and as an adjunct to colonoscopy every 5 years in persons with first-degree relatives who had colorectal cancer. RESULTS: In the base case, the incremental cost per life-year saved with a COX-2 inhibitor alone compared with no screening was 233,300 dollars in persons at average risk of colorectal cancer and 56,700 dollars in persons with 2 first-degree relatives who had the disease. Chemoprevention was both less effective and more costly than screening. The incremental cost per life-year saved with a COX-2 inhibitor as an adjunct to screening was 823,800 dollars in persons at average risk and 404,700 dollars in persons with 2 first-degree relatives who had colorectal cancer. Combining a COX 2 inhibitor with less frequent screening was not as cost-effective as screening at currently recommended intervals. Cost-effectiveness estimates were highly sensitive to the cost of COX-2 inhibitors and their effect on the risk of cancer. CONCLUSION: Chemoprevention of colorectal cancer with COX-2 inhibitors is likely to incur substantially higher costs per life-year saved than are currently recommended screening strategies. COX-2 inhibitor use as an adjunct to screening may increase life expectancy, although at prohibitive costs, and is unlikely to result in less frequent screening. PMID- 12753879 TI - Intensive smoking cessation counseling versus minimal counseling among hospitalized smokers treated with transdermal nicotine replacement: a randomized trial. AB - PURPOSE: To determine whether an intensive cognitive-behavioral intervention begun during hospitalization when combined with transdermal nicotine replacement therapy is more effective than a minimal counseling intervention combined with transdermal nicotine replacement therapy in helping inpatients to quit smoking. METHODS: A total of 223 patients who smoked were enrolled in a hospital-based randomized smoking cessation trial at the San Francisco Veterans Affairs Medical Center. One hundred and seven participants (48%) received intensive counseling and outpatient telephone follow-up; 116 participants (52%) received minimal counseling. All study participants received 2 months of transdermal nicotine replacement therapy. We determined 6-month quit rates by self-report and measured saliva cotinine levels or obtained proxy reports to confirm self-reported smoking cessation at 12 months. Analyses adjusted for baseline differences in the distribution of coronary disease. RESULTS: At 6 months, 35% (36/103) of the intensive intervention group reported quitting, compared with 21% (23/109) of the comparison group (relative risk [RR] = 1.7; 95% confidence interval [CI]: 1.1 to 2.7). At 12 months, the self-reported quit rate was 33% (33/99) in the intensive intervention group versus 20% (21/103) in the comparison group (RR = 1.7; 95% CI: 1.1 to 2.7). Based on biochemical or proxy confirmation, 29% (30/102) in the intensive intervention group versus 20% (21/107) in the comparison group quit smoking at 12 months (RR = 1.6; 95% CI: 0.96 to 2.5). CONCLUSION: Hospital initiated smoking cessation interventions that include transdermal nicotine replacement therapy can improve long-term quit rates. PMID- 12753880 TI - Nephrogenic fibrosing dermopathy: a novel cutaneous fibrosing disorder in patients with renal failure. AB - BACKGROUND: Nephrogenic fibrosing dermopathy is a newly recognized cutaneous fibrosing disorder marked by the acute onset of induration involving the upper and lower limbs in patients with acute or chronic renal failure. The etiology, pathogenesis, associated clinical conditions (other than renal failure), and ultimate course have not been defined in the few cases studied. Presently, there is no effective treatment, and the condition persists in most patients. METHODS: Clinical and histopathologic data on 13 patients from our institution with the diagnosis of nephrogenic fibrosing dermopathy were reviewed. Several clinical and laboratory parameters were examined to see if any were consistently associated with the disease. Biopsy specimens were analyzed to determine if there was a pattern to the evolution of fibrosis in these patients. RESULTS: All 13 patients had renal failure before disease onset: 8 were undergoing chronic hemodialysis, 2 were undergoing chronic peritoneal dialysis, and 3 with acute renal failure had never undergone dialysis before the development of dermopathy. Most patients had other serious underlying medical conditions. Many patients were taking erythropoietin, cyclosporine, or both before the onset of disease. In transplant patients, no histocompatibility antigens were found to be associated with the disease. There were various laboratory abnormalities, but none were consistently associated with the condition. In skin biopsy specimens taken 7 to 180 days after disease onset, there were histopathologic changes suggestive of a tissue reaction to injury, as well as the development of smooth muscle actin-positive myofibroblasts. CONCLUSION: Nephrogenic fibrosing dermopathy is a novel cutaneous fibrosing disorder that is distinguished from other sclerosing or fibrosing skin disorders by distinctive clinical and histopathologic findings occurring in the setting of renal failure. There were no additional clinical risk factors or laboratory findings common to the 13 patients studied, other than renal failure. The resemblance to a tissue injury reaction and the presence of myofibroblasts in the tissue specimens suggest that fibrogenic cytokines may be involved in the evolution of the disease. PMID- 12753881 TI - Substance use and mental health correlates of nonadherence to antiretroviral medications in a sample of patients with human immunodeficiency virus infection. AB - PURPOSE: Mental health and substance use problems are common among patients infected with human immunodeficiency virus (HIV) and may impede adherence to antiretroviral regimens. This study investigated associations of antiretroviral medication nonadherence with specific types of psychiatric disorders and drug use, and varying levels of alcohol use. METHODS: Data were drawn from a survey of a national probability sample of 2267 (representing 181,557) adults enrolled in the HIV Cost and Services Utilization Study. This study focused on 1910 patients who reported their antiretroviral medication adherence during the past week. RESULTS: Patients with depression (odds ratio [OR] = 1.7; 95% confidence interval [CI]: 1.3 to 2.3), generalized anxiety disorder (OR = 2.4; 95% CI: 1.2 to 5.0), or panic disorder (OR = 2.0; 95% CI: 1.4 to 3.0) were more likely to be nonadherent than those without a psychiatric disorder. Nonadherence was also associated with use of cocaine (OR = 2.2; 95% CI: 1.2 to 3.8), marijuana (OR = 1.7; 95% CI: 1.2 to 2.3), amphetamines (OR = 2.3; 95% CI: 1.2 to 4.2), or sedatives (OR = 1.6; 95% CI: 1.0 to 2.4) in the previous month. Compared with patients who did not drink, those who were moderate (OR = 1.6; 95% CI: 1.3 to 2.0), heavy (OR = 1.7; 95% CI: 1.3 to 2.3), or frequent heavy (OR = 2.7; 95% CI: 1.7 to 4.5) drinkers were more likely to be nonadherent. These associations could not be explained by demographic, clinical, and treatment factors. CONCLUSION: These findings suggest the need for screening and treatment for mental health and substance use problems among HIV-positive patients to improve adherence to antiretroviral medications. PMID- 12753882 TI - Effects of physician-related factors on adult asthma care, health status, and quality of life. AB - PURPOSE: To study the association of physician characteristics, the characteristics of their practice settings, patient mix, and reported frequency of prescribing asthma medication with patients' health status and health-related quality of life in asthma. METHODS: We conducted a mail-back survey of physicians (n = 147) that included demographic characteristics, practice and training characteristics, and reported prescribing frequencies for common asthma treatments. We also conducted structured telephone interviews with 317 of their patients, assessing demographic characteristics, health status (as measured by the Short Form-12 [SF-12] physical component score), and asthma-specific quality of life (as measured by the Marks questionnaire). RESULTS: In adjusted analyses, pulmonary specialists were more likely to report using leukotriene modifiers (odds ratio [OR] = 4.7; 95% confidence interval [CI]: 1.2 to 18) and theophylline (OR = 3.0; 95% CI: 1.0 to 9.0) in adult patients with asthma. Working in a practice of >75% health maintenance organization (HMO)- or preferred provider organization (PPO)-insured patients was associated with a lower likelihood of prescribing leukotriene modifiers (OR = 0.1; 95% CI: 0.01 to 0.5). Adjusting for patient demographic characteristics and steroid dependence, physician prescribing tendencies were not associated with patients' perceived health status or quality of life. Although an HMO- or PPO-predominant practice was associated with better physical health status (mean difference in SF-12 physical component score, 3.1; 95% CI: 0.05 to 6.2; P = 0.05), there was no statistical association with quality of life. CONCLUSION: The characteristics of physicians, their practices, and the asthma medication prescribing strategies that they adopt are not strongly associated with patients' perceived outcomes. PMID- 12753883 TI - Comparative effects of angiotensin-converting enzyme inhibitors and angiotensin receptor blockers on blood pressure and the kidney. AB - Many clinicians are uncomfortable about using angiotensin-converting enzyme (ACE) inhibitors or angiotensin II type 1 receptor blockers (AT(1)-blockers) to treat patients with renal disease because of concerns about increasing serum creatinine levels. However, the benefits of these medications, particularly their efficacy in slowing the progression of renal disease, outweigh such concerns. ACE inhibitors are effective in patients with type 1 diabetes and renal disease, as well as in those with nondiabetic renal disease and proteinuria >0.5 g/d. AT(1) blockers slow the progression of diabetic nephropathy in patients with type 2 diabetes. Although these classes of medications should not be used in patients with severe renal insufficiency (e.g., glomerular filtration rate <20 mL/min), they may be beneficial in patients with mild-to-moderate renal insufficiency. Nonetheless, caution should be exercised in those with a glomerular filtration rate <30 mL/min, and serum creatinine and potassium levels should be checked approximately 1 week after starting treatment. There is also evidence suggesting that these medications lead to greater reductions in blood pressure and proteinuria when used in combination than when alone. The purpose of this paper is to review the mechanisms of action of these two classes of medication, as well as the experimental and clinical evidence that they slow the progression of renal disease. PMID- 12753884 TI - Influence of the menstrual cycle on the timing of acute coronary events in premenopausal women. PMID- 12753885 TI - Unsuspected bacterial suppurative disease of the airways presenting as chronic cough. PMID- 12753886 TI - Is inflammation the critical factor linking vulnerable coronary plaques to clinical coronary disease? PMID- 12753887 TI - Improving the laboratory diagnosis of heparin-induced thrombocytopenia. PMID- 12753888 TI - Myocardial infarction during menses: lessons from trials and errors. PMID- 12753889 TI - Cases from the Osler medical service at Johns Hopkins University. AB - A 56-year-old black woman with diabetes mellitus was admitted for hypoglycemia and confusion. Her past medical history included breast cancer, for which she had undergone a left lumpectomy and then mastectomy for in-breast recurrence. Her oral intake had decreased during the past month because of increasing discomfort from left-sided chest pain. During this period, she continued to take pioglitazone for diabetes at her originally prescribed dose. The patient's mental status improved quickly after taking orange juice and intravenous glucose, but the chest pain persisted. The pain, which was described as an ache along the left costal margin, increased with palpation, deep inspiration, or coughing. She had recently presented with similar complaints at another hospital where she had been prescribed a muscle relaxant that provided no relief from the pain. She also reported a 14-lb weight loss during the previous 3 months, as well as fatigue, weakness, and aches in her legs and arms. She denied fevers, chills, sweats, abdominal pain, nausea, or recent trauma. Laboratory values at the time of admission were: calcium, 11.8 mg/dL; total protein, 11.1 mg/dL; albumin, 3.2 g/dL; creatinine, 1.0 mg/dL; and hematocrit, 29.3%, with a mean corpuscular volume of 89.3. Chest radiography revealed a lytic lesion in the left lateral fourth rib and left humerus (). Serum and urine protein electrophoresis revealed a monoclonal spike in the gamma region consistent with monoclonal gammopathy. The serum spike was quantified at 3.78 g/dL. A skeletal survey showed many small well defined lytic lesions in the skull (with one 1.5-cm lytic lesion in the upper posterior parietal bone), arms, and legs. A bone scan showed multiple foci of increased uptake in the right and left ribs as well as the proximal portion of the left femur. The peripheral blood smear revealed rouleaux formation () and plasma cells (). What is the diagnosis? PMID- 12753890 TI - The nutcracker phenomenon accompanied by renin-dependent hypertension. PMID- 12753891 TI - Mucormycosis of the tongue in a patient with acute lymphoblastic leukemia: a possible relation with use of a tongue depressor. PMID- 12753892 TI - Effects of calcium supplementation on serum lipid levels in postmenopausal women. PMID- 12753893 TI - Reforming the medical professional liability insurance system. PMID- 12753894 TI - Is a Q-cycle-like mechanism operative in dihaemic succinate:quinone and quinol:fumarate oxidoreductases? AB - Succinate:quinone (SQR) and quinol:fumarate oxidoreductases (QFR) are members of the same enzyme family. These are membrane bound enzymes anchored to the membrane by one or two subunits that may contain two, one or no haems. For the dihaemic enzymes the electron pathway from the flavin at the catalytic centre to the quinones remains to be established. Taking into account that the two haems are located on opposite sites of the membrane, and the possible presence of two quinone binding sites, also located on opposite sides of the membrane, we re hypothesise the presence of a Q-cycle type mechanism in these enzymes. Such a mechanism can explain an active functional role for two haems and two quinone binding sites, allowing SQR to conserve energy. With this testable hypothesis we intend to challenge the discussion and drive further experimentation to unravel the functional mechanism of SQRs and QFRs. PMID- 12753895 TI - Thyroid hormone and uncoupling proteins. AB - Thyroid hormone (TH/T3) exerts many of its effects on energy metabolism by affecting gene transcription. However, although this is an important target for T3, only a limited number of T3-responsive genes have been identified and studied. Among these, the genes for uncoupling proteins (UCPs) have attracted the interest of scientists. Although the role of UCP1 seems quite well established, uncertainty surrounds the physiological function of the recently discovered UCP1 analogs, UCP2 and UCP3. The literature suggests that T3 affects both the expression and the activity of each of these UCPs but further studies are needed to establish whether the mechanisms activated by the hormone are the same. Recently, because of their larger range of expression, much attention has been devoted to UCP2 and UCP3. Most detailed studies on the involvement of these proteins as mediators of the effects of T3 on metabolism have focused on UCP3 because of its expression in skeletal muscle. T3 seems to be unique in having the ability to stimulate the expression and activity of UCP3 and this may be related to the capacity of T3 to activate the integrated biochemical processes linked to UCP activity, such as those related to fatty acids, coenzyme Q and free radicals. PMID- 12753896 TI - Structural analysis of mycobacterial and murine hsp60 epitopes in complex with the class I MHC molecule H-2Db. AB - The decameric peptide SALQNAASIA from the Mycobacterium bovis heat shock protein (hsp) 60 is recognized by the murine T-cell receptor UZ-3-4 in complex with the murine class I major histocompatibility complex molecule H-2D(b). This T-cell receptor cross-reacts with the H-2D(b)-bound non-homologous decameric peptide KDIGNIISDA from the murine hsp60, but does not recognize the nonameric mycobacterial peptide SALQNAASI. Cross-recognition of the KDIGNIISDA/H-2D(b) complex induces autoimmune pathology in immunodeficient mice. We solved the X-ray crystal structure of the SALQNAASIA/H-2D(b) complex at 3.0 A resolution, and we modelled the KDIGNIISDA and SALQNAASI peptides in the H-2D(b) binding site. The structural analysis of the H-2D(b)-bound hsp60 epitopes offers insight into T cell receptor cross-reactivity. PMID- 12753897 TI - Identification of two serine residues important for p53 DNA binding and protein stability. AB - The p53 core DNA binding domain has been implied in Mdm2-mediated protein degradation. Here we show that the substitution of the serine residues 116 and 127 with alanine residues (S116/127A) has no effect on p53 DNA binding and protein stability. However, the substitution of the serine residues with the aspartic acid (S116/127D) abolished p53 DNA binding and led to protein stabilization. Importantly, we have shown that S116/127D exhibits a structural mutant conformation that results in a loss of p53-dependent transcription and Mdm2-mediated protein degradation. PMID- 12753898 TI - Identification and characterisation of human aldose 1-epimerase. AB - Aldose 1-epimerase or mutarotase (EC 5.1.3.3) is a key enzyme of carbohydrate metabolism catalysing the interconversion of the alpha- and beta-anomers of hexose sugars such as glucose and galactose. We identified an open reading frame in the human genome (BC014916) which has high sequence similarity to previously identified bacterial aldose 1-epimerases. This sequence was cloned into a bacterial expression vector, and expressed and purified from this source. Enzyme assays show that the protein has aldose 1-epimerase activity and exhibits a preference for galactose over glucose. Site-directed mutagenesis confirmed the involvement of three residues involved in catalysis and substrate binding. PMID- 12753899 TI - Dexamethasone reverses TGF-beta-mediated inhibition of primary rat preadipocyte differentiation. AB - Dexamethasone and transforming growth factor-beta (TGF-beta) show contrary effects on differentiation of adipocytes. Dexamethasone stimulates adipocyte differentiation whereas TGF-beta inhibits it. In the present study, we investigated whether dexamethasone could reverse the TGF-beta-mediated inhibition of preadipocyte differentiation. Primary rat preadipocytes, obtained from Sprague Dawley rats, were pretreated with dexamethasone in the presence or absence of TGF beta, prior to the induction of differentiation. Co-treatment of dexamethasone and TGF-beta before inducing differentiation reversed the TGF-beta-mediated inhibition of preadipocyte differentiation. In order to elucidate the mechanism by which dexamethasone reversed the effect of TGF-beta on the inhibition of preadipocyte differentiation, the expression of CCAAT/enhancer binding protein alpha (C/EBPalpha) and peroxisome proliferator-activated receptor gamma (PPARgamma) was examined. Dexamethasone increased C/EBPalpha and PPARgamma expression in the absence of TGF-beta and also recovered the TGF-beta-mediated suppression of C/EBPalpha expression in preadipocytes. Its effect was sustained in differentiated adipocytes as well. However, those effects were not observed in 3T3-L1 preadipocytes or differentiated adipocytes. These results indicate that dexamethasone reverses the TGF-beta-mediated suppression of adipocyte differentiation by regulating the expression of C/EBPalpha and PPARgamma, which is dependent on the cellular context. PMID- 12753900 TI - The Methanocaldococcus jannaschii protein Mj0968 is not a P-type ATPase. AB - The Methanocaldococcus jannaschii (formerly Methanococcus jannaschii) protein Mj0968 has been reported to represent a soluble P-type ATPase [Ogawa et al., FEBS Lett. 471 (2000) 99-102]. In this study, we report that the heterologously expressed Mj0968-His(10) protein exhibits high rates of phosphatase activity, whereas only very low ATPase activity was measured. Replacement of the aspartate residue in the DSAGT motif (D7A), which becomes phosphorylated during the reaction cycle of P-type ATPases, does not affect the V(max), but only the K(M) of the reaction. Labeling studies with [gamma-(32)P]ATP and [alpha-(32)P]ATP revealed that the previously reported labeling experiments [Ogawa et al., 2000] do not necessarily show phosphorylation of Mj0968, but rather point to ATP binding. Binding studies with trinitrophenyl adenosine nucleotides showed low apparent K(d) values for those molecules. These results provide evidence that the native function of Mj0968 seems to be that of a phosphatase, rather than that of an ATP-hydrolyzing enzyme. PMID- 12753901 TI - pH opposite effects on synthesis of dinucleoside polyphosphates and on oxidation reactions catalyzed by firefly luciferase. AB - Previous results have shown that an oxidizing product of firefly luciferin, dehydroluciferyl-adenylate, is the main intermediate in the process of synthesis of dinucleoside polyphosphates catalyzed by firefly luciferase (EC 1.13.12.7). However, we have found that the pH effects on the luciferase oxidizing processes and on the synthesis of dinucleoside polyphosphate are opposite: acidic assay media enhance the synthesis of dinucleoside polyphosphate and inhibit the oxidizing processes. The reason for this apparent contradiction lies on the activation effect of low pH on the adenylate transfer reaction from dehydroluciferyl-adenylate to the acceptor nucleotide. PMID- 12753902 TI - Transcriptional effects of the signal transduction protein P(II) (glnB gene product) on NtcA-dependent genes in Synechococcus sp. PCC 7942. AB - P(II) proteins signal the cellular nitrogen status in numerous bacteria, and in cyanobacteria P(II) is subjected to serine phosphorylation when the cells experience a high C to N balance. In the unicellular cyanobacterium Synechococcus sp. PCC 7942, the P(II) protein (glnB gene product) is known to mediate the ammonium-dependent inhibition of nitrate and nitrite uptake. The analysis of gene expression through RNA/DNA hybridization indicated that a P(II)-null mutant was also impaired in the induction of NtcA-dependent, nitrogen assimilation genes amt1 (ammonium permease), glnA (glutamine synthetase) and nir (nitrite reductase), as well as of the N-control gene ntcA, mainly under nitrogen deprivation. This gene expression phenotype of the glnB mutant could be complemented by wild-type P(II) protein or by modified P(II) proteins that cannot be phosphorylated and mimic either the phosphorylated (GlnB(S49D) and GlnB(S49E)) or unphosphorylated (GlnB(S49A)) form of P(II). However, strains carrying the GlnB(S49D) and GlnB(S49E) mutant proteins exhibited higher levels of expression of nitrogen-regulated genes than the strains carrying the wild-type P(II) or the GlnB(S49A) protein. PMID- 12753903 TI - The A-type ATP synthase subunit K of Methanopyrus kandleri is deduced from its sequence to form a monomeric rotor comprising 13 hairpin domains. AB - The ntpK gene of the archaeon Methanopyrus kandleri encodes the equivalent of the c subunit of ATP synthase. The gene product contains 1021 residues and consists of 13 homologous domains, each one corresponding to a single helical hairpin. The amino acid sequence of the domains is highly conserved, ranging between 50 and 80% sequence identity. Each of the 13 domains contains a conserved Gln and Glu residue in the N- and C-terminal helix, respectively, both of which are believed to be involved in cation binding. The protein is likely to form the monomeric rotor of the ATP synthase that consists of 13 hairpin domains. PMID- 12753904 TI - Short interfering RNA-directed inhibition of hepatitis B virus replication. AB - RNA interference (RNAi) is the process by which double-stranded RNA directs sequence-specific degradation of mRNA. In mammalian cells, RNAi can be triggered by 21-nucleotide duplexes of short interfering RNA (siRNA). We examined effects of siRNA on hepatitis B virus (HBV) replication. Human hepatoma cells were transfected with HBV DNA and siRNA against HBV-pregenome RNA. Transfection experiments demonstrated that the siRNA reduced the amount of HBV-pregenome RNA and resulted in reduction of the levels of replicative intermediates and viral protein. Our results indicate that siRNA-mediated gene silencing inhibits HBV replication through suppression of viral RNA, which may be useful as a potential therapeutic modality. PMID- 12753905 TI - The A20-binding protein ABIN-2 exerts unexpected function in mediating transcriptional coactivation. AB - The human ABIN-2 was originally identified as an A20-associating cytosolic protein to block NF-kappaB activation induced by various stimuli. Here we report that ABIN-2 has the potential to enter the nucleus and plays a role in mediating transcriptional activation in both yeast and mammalian cells. The Gal4BD-ABIN-2 fusion protein is able to drive the expression of the GAL4-responsive reporter gene in yeast efficiently without the need of the Gal4p activation domain, suggesting that ABIN-2 functions as a transcriptional coactivator and facilitates transcription in yeast. In contrast to the activity in yeast, however, only the C terminal fragment of ABIN-2 exerts the transactivating activity in mammalian cells but not the full-length ABIN-2 protein. This observation has led to the identification of the N-terminal 195 amino acids of ABIN-2 as a regulatory domain, which retains the full-length ABIN-2 in the cytoplasm of mammalian cells and thus cannot transactivate. We have also found that BAF60a, a component of chromatin-remodeling complex, interacts with ABIN-2 by the yeast two-hybrid analysis. Together, our results suggest that the nuclear ABIN-2 defines a novel transcriptional coactivator and acts presumably by recruiting a chromatin remodeling complex to the site of the target gene. PMID- 12753906 TI - Interaction of Sp1 transcription factor with HIV-1 Tat protein: looking for cellular partners. AB - The Tat protein of human immunodeficiency virus type 1 (HIV-1) trans-activates HIV-1 transcription by functionally interacting with a number of cellular proteins, among which the Sp1 transcription factor. We recently demonstrated that Tat does not directly interact with Sp1 either in vitro or in vivo, and we suggested that other protein(s) could indirectly mediate Tat-Sp1 interaction. In keeping, here we showed that addition of HeLa cell nuclear extracts to purified Tat and Sp1 proteins allows the formation of a Tat/Sp1 complex in in vitro binding assays. In an attempt to identify the partner(s) that bridge Tat and Sp1, we developed a yeast multi-protein system, in which cellular proteins recently shown to play a relevant role in Tat function, namely TATA box-binding protein, cyclin T1, CDK9, and cyclin T1/CDK9 complex, were coexpressed, individually or in pair-wise combination, with Tat and Sp1 hybrids. We demonstrated that none of these candidate partners bridges Tat and Sp1. However, our yeast multi-protein system, which allows simple and rapid detection of interactions among up to four proteins, will be most helpful to further dissect the interaction of Tat and Sp1 with other candidate partners that participate in the assembly of transcriptionally active complexes at the HIV-1 LTR. PMID- 12753907 TI - gamma-MSH increases intracellular cAMP accumulation and GnRH release in vitro and LH release in vivo. AB - The roles of the melanocortin 3 receptor (MC3-R) and its agonist, gamma(2) melanocyte-stimulating hormone (gamma(2)-MSH) in the regulation of the hypothalamo-pituitary-gonadal (HPG) axis are poorly understood. Here we show gamma(2)-MSH stimulated intracellular cAMP accumulation and gonadotrophin releasing hormone (GnRH) secretion in the immortalised GnRH cell line GT(1)-7. The MC3/4-R antagonist Agrp blocked these actions. Reverse transcriptase polymerase chain reaction demonstrated GT(1)-7 cells express MC3-R mRNA. gamma(2) MSH also stimulated GnRH release from hypothalamic explants. In vivo, gamma(2) MSH administration into the medial preoptic area significantly increased plasma luteinising hormone. MC3-R and gamma(2)-MSH may modulate the HPG axis. PMID- 12753908 TI - A molecular mechanism for the low-pH stability of sialidase activity of influenza A virus N2 neuraminidases. AB - Four human pandemic influenza A virus strains isolated in 1957 and 1968, but not most of the epidemic strains isolated after 1968, possess sialidase activity under low-pH conditions. Here, we used cell-expressed neuraminidases (NAs) to determine the region of the N2 NA that is associated with low-pH stability of sialidase activity. We found that consensus amino acid regions responsible for low-pH stability did not exist in pandemic NAs but that two amino acid substitutions in the low-pH-stable A/Hong Kong/1/68 (H3N2) NA and a single substitution in the low-pH-unstable A/Texas/68 (H2N2) NA resulted in significant change in low-pH stability. PMID- 12753909 TI - Heregulin and HER2 signaling selectively activates c-Src phosphorylation at tyrosine 215. AB - To elucidate the molecular mechanisms by which human epidermal growth factor receptor/heregulin (HER2/HRG) influence the migratory potential of breast cancer cells, we have used phospho-specific antibodies against c-Src kinase and focal adhesion kinase (FAK). This study establishes that HER2/HRG signaling selectively upregulates Tyr phosphorylation of c-Src at Tyr-215 located within the SH2 domain, increases c-Src kinase activity and selectively upregulates Tyr phosphorylation of FAK at Tyr-861. HER2-overexpressing tumors showed increased levels of c-Src phosphorylation at Tyr-215. These findings suggest that HER2/HRG influence metastasis of breast cancer cells through a novel signaling pathway involving phosphorylation of FAK tyrosine 861 via activation of c-Src tyrosine 215. PMID- 12753910 TI - Initiation factor eIF2B not p70 S6 kinase is involved in the activation of the PI 3K signalling pathway induced by the v-src oncogene. AB - Our data show that in hamster fibroblasts transformed by Rous sarcoma virus (RSV), the phosphoinositide 3'-kinase (PI-3K)/Akt/glycogen synthase kinase 3 antiapoptotic pathway is upregulated and involved in increased protein synthesis through activation of initiation factor eIF2B. Upon inhibition of PI-3K by wortmannin, phosphorylation of 70-kDa ribosomal protein S6 kinase (p70 S6k) and its physiological substrate, ribosomal protein S6, decreased in the non transformed cells but not in RSV-transformed cells. Thus PI-3K, which is thought to be involved in regulation of p70 S6k, signals to p70 S6k in normal fibroblasts, but it does not appear to be an upstream effector of p70 S6k in fibroblasts transformed by v-src oncogene, suggesting that changes in the PI-3K signalling pathway upstream of p70 S6k are induced by RSV transformation. PMID- 12753911 TI - Characterization of thioredoxin y, a new type of thioredoxin identified in the genome of Chlamydomonas reinhardtii. AB - The sequencing of the Arabidopsis genome revealed a multiplicity of thioredoxins (TRX), ubiquitous protein disulfide oxido-reductases. We have analyzed the TRX family in the genome of the unicellular green alga Chlamydomonas reinhardtii and identified eight different thioredoxins for which we have cloned and sequenced the corresponding cDNAs. One of these TRXs represents a new type that we named TRX y. This most probably chloroplastic TRX is highly conserved in photosynthetic organisms. The biochemical characterization of the recombinant protein shows that it exhibits a thermal stability profile and specificity toward target enzymes completely different from those of TRXs characterized so far. PMID- 12753912 TI - Microwave radiation can alter protein conformation without bulk heating. AB - Exposure to microwave radiation enhances the aggregation of bovine serum albumin in vitro in a time- and temperature-dependent manner. Microwave radiation also promotes amyloid fibril formation by bovine insulin at 60 degrees C. These alterations in protein conformation are not accompanied by measurable temperature changes, consistent with estimates from field modelling of the specific absorbed radiation (15-20 mW kg(-1)). Limited denaturation of cellular proteins could explain our previous observation that modest heat-shock responses are induced by microwave exposure in Caenorhabditis elegans. We also show that heat-shock responses both to heat and microwaves are suppressed after RNA interference ablating heat-shock factor function. PMID- 12753913 TI - Chemical structure and immunobiological activity of lipid A from Prevotella intermedia ATCC 25611 lipopolysaccharide. AB - The novel chemical structure and immunobiological activities of Prevotella intermedia ATCC 25611 lipid A were investigated. A lipopolysaccharide (LPS) preparation of P. intermedia was extracted using a phenol-chloroform-petroleum ether method, after which its purified lipid A was prepared by weak acid hydrolysis followed by chromatographic separations. The lipid A structure was determined by mass spectrometry and nuclear magnetic resonance to be a diglucosamine backbone with a phosphate at the 4-position of the non-reducing side sugar, as well as five fatty acids containing branched long chains. It was similar to that of Bacteroides fragilis and Porphyromonas gingivalis, except for the phosphorylation site. P. intermedia lipid A induced weaker cytokine production and NF-kappaB activation in murine cells via Toll-like receptor (TLR) 4 as compared to Escherichia coli synthetic lipid A (compound 506). Our results indicate that P. intermedia lipid A activates cells through a TLR4-dependent pathway similar to E. coli-type lipid A, even though these have structural differences. PMID- 12753914 TI - Stimulation of Na+/Mg2+ antiport in rat erythrocytes by intracellular Cl-. AB - Mg(2+) efflux from rat erythrocytes was measured in NaCl, NaNO(3), NaSCN and Na gluconate medium. Substitution of extracellular and intracellular Cl(-) with the permeant anions NO(3)(-) and SCN(-) reduced Mg(2+) efflux via Na(+)/Mg(2+) antiport. After substitution of extracellular Cl(-) with the non-permeant anion gluconate, Mg(2+) efflux was not significantly reduced. In Na gluconate medium, an influence of the changed membrane potential and intracellular pH on Mg(2+) efflux could be excluded. The results indicate the existence of Cl(-)-independent Na(+)/Mg(2+) antiport and of Na(+)/Mg(2+) antiport stimulated by intracellular Cl(-). Intracellular Cl(-), as determined by means of (36)Cl(-), was found to stimulate Na(+)/Mg(2+) antiport through a cooperative effect according to a sigmoidal kinetics. The Hill coefficient for intracellular Cl(-) amounted to 1.4 1.8, indicating that two intracellular Cl(-) may be simultaneously active. With respect to specificity, Cl(-) was most effective, followed by Br(-), J(-), and F( ). Stimulation of Na(+)/Mg(2+) antiport by intracellular Cl(-) together with intracellular Mg(2+) may play a role during deoxygenation of erythrocytes and in essential hypertension. PMID- 12753915 TI - Channel induction by palytoxin in yeast cells expressing Na+,K+-ATPase or its chimera with sarco/endoplasmic reticulum Ca2+-ATPase. AB - Palytoxin (PTX) induces a cation channel through interaction with Na(+),K(+) ATPase. It is unclear how this action relates to the enzyme catalytic activity. We examined whether the action of PTX depends on the catalytic domain specific for Na(+),K(+)-ATPase. Wild-type Na(+),K(+)-ATPase alpha-subunit (NNN) or its chimera (NCN), in which the catalytic domain was replaced with that of sarcoplasmic/endoplasmic reticulum Ca(2+)-ATPase, was co-expressed with beta subunit in the yeast Saccharomyces cerevisiae. PTX (0.1-100 nM) increased K(+) efflux in NNN- or NCN-transfected cells to a similar degree but not in non transfected cells. When ouabain-resistant NNN and NCN were expressed, PTX also increased K(+) efflux. Ouabain inhibited the effect of PTX in NNN or NCN cells but not in ouabain-resistant cells. These data suggest that the channel-forming action of PTX does not depend on the catalytic domain species. PMID- 12753916 TI - Interaction of bilirubin and biliverdin with reactive nitrogen species. AB - Bilirubin (BR) and biliverdin (BV), two metabolites produced during haem degradation by haem oxygenase, possess strong antioxidant activities toward peroxyl radical, hydroxyl radical and hydrogen peroxide. Considering the importance attributed to nitric oxide (NO) and its congeners in the control of physiological and pathophysiological processes, we examined the interaction of BR and BV with NO and NO-related species in vitro. Exposure of BR and BV to agents that release NO or nitroxyl resulted in a concentration- and time-dependent loss of BR and BV, as assessed by high performance liquid chromatography. Peroxynitrite, a strong oxidant derived from the reaction of NO with superoxide anion, also showed high reactivity toward BR and BV. The extent of BR and BV consumption largely depended on the NO species being analysed and on the half lives of the pharmacological compounds considered. Of major importance, BR and BV decomposition occurred also in the presence of pure NO under anaerobic conditions, confirming the ability of bile pigments to scavenge the gaseous free radical. Increasing concentrations of thiols prevented BR consumption by nitroxyl, indicating that bile pigments and thiol groups can compete and/or synergise the cellular defence against NO-related species. In view of the high inducibility of haem oxygenase-1 by NO-releasing agents in different cell types, the present findings highlight novel anti-nitrosative characteristics of BR and BV suggesting a potential function for bile pigments against the damaging effects of uncontrolled NO production. PMID- 12753917 TI - In vivo stabilization of nuclear retinoid X receptor alpha in the presence of peroxisome proliferator-activated receptor alpha. AB - Retinoid X receptor alpha (RXRalpha) can reveal diverse functions through forming a heterodimer with peroxisome proliferator-activated receptor alpha (PPARalpha). However, the mechanism of regulation of the cellular RXRalpha level is unclear. Thus, quantitative change of RXRalpha was investigated in mouse liver. Nuclear RXRalpha level was constitutively lower in PPARalpha-null mice than in wild-type mice. The level was also increased by clofibrate treatment in wild-type mice without a concomitant increase of RXRalpha mRNA, but not in PPARalpha-null mice. Pulse chase experiments demonstrated that the presence of PPARalpha and its activation by ligands significantly affected the stability of nuclear RXRalpha. These findings suggest a novel regulatory mechanism of nuclear RXRalpha in vivo. PMID- 12753918 TI - Signal-induced ubiquitination of p57(Kip2) is independent of the C-terminal consensus Cdk phosphorylation site. AB - The cyclin-dependent kinase inhibitor p57(Kip2) is required for normal mouse embryonic development. p57(Kip2) consists of four structurally distinct domains in which the conserved C-terminal nuclear targeting domain contains a putative Cdk phosphorylation site (Thr(342)) that shares a great similitude in the adjacent sequences with p27(Kip1) but not with p21(Cip1). Phosphorylation on Thr(187) has been shown to promote degradation of p27(Kip1). Although there is sequence homology between the C-terminal part of p27(Kip1) and p57(Kip2), we show that the ubiquitination and degradation of p57(Kip2) are independent of Thr(342). In contrast a destabilizing element located in the N-terminal is implicated in p57(Kip2) destabilization. PMID- 12753919 TI - PAK interacts with NCK and MLK2 to regulate the activation of jun N-terminal kinase. AB - The p21-GTPase activated kinase, PAK1, and the mixed lineage kinase, MLK2, have been implicated in the activation of jun N-terminal kinase (JNK). However, the role of PAK1 in JNK activation is still not understood. Here we show that over expression of the SH3-SH2 adapter Nck 'squelches' JNK activation but this squelching is relieved by over-expression of PAK1. In turn, PAK1 squelches activation of JNK by MLK2 and these kinases interact via their catalytic domains. The data suggest that PAK1 recruits MLK2 to an activated receptor via the adapter Nck, but cannot itself induce activation of the JNK cascade. PMID- 12753920 TI - The gene encoding glutathione-dependent formaldehyde dehydrogenase/GSNO reductase is responsive to wounding, jasmonic acid and salicylic acid. AB - It has recently been discovered that glutathione-dependent formaldehyde dehydrogenase (FALDH) exhibits a strong S-nitrosoglutathione reductase activity. Plants use NO and S-nitrosothiols as signaling molecules to activate defense mechanisms. Therefore, it is interesting to investigate the regulation of FALDH by mechanical wounding and plant hormones involved in signal transduction. Our results show that the gene encoding FALDH in Arabidopsis (ADH2) is down-regulated by wounding and activated by salicylic acid (SA). In tobacco, FALDH levels and enzymatic activity decreased after jasmonate treatment, and increased in response to SA. This is the first time that regulation of FALDH in response to signals associated with plant defense has been demonstrated. PMID- 12753921 TI - Regulatory mechanism of Cordyceps sinensis mycelium on mouse Leydig cell steroidogenesis. AB - We demonstrate the mechanism by which Cordyceps sinensis (CS) mycelium regulates Leydig cell steroidogenesis. Mouse Leydig cells were treated with forskolin, H89, phorbol 12-myristate 13-acetate, staurosporine, or steroidogenic enzyme precursors with or without 3 mg/ml CS; then testosterone production was determined. H89, but not phorbol 12-myristate 13-acetate or staurosporine, decreased CS-treated Leydig cell steroidogenesis. CS inhibited Leydig cell steroidogenesis by suppressing the activity of P450scc enzyme, but not 3beta hydroxysteroid dehydrogenase, 17alpha-hydroxylase, 20alpha-hydroxylase, or 17beta hydroxysteroid dehydrogenase enzymes. Thus, CS activated the cAMP-protein kinase A signal pathway, but not protein kinase C, and attenuated P45scc enzyme activity to reduce human chorionic gonadotropin-stimulated steroidogenesis in purified mouse Leydig cells. PMID- 12753922 TI - Oxidation of nuclear thioredoxin during oxidative stress. AB - Thioredoxin 1 (Trx1) is a key redox control system within the nucleus, yet little is known about the sensitivity of nuclear Trx1 to oxidative stress. The present study compared oxidant-induced changes in the redox states of nuclear Trx1, cytoplasmic Trx1, and cellular glutathione (GSH). Nuclear Trx1 was more reducing than cytoplasmic Trx1 and cellular GSH in proliferating cells. tert Butylhydroperoxide caused an increase in the total amount of nuclear Trx1, but this was accompanied by a 60 mV oxidation. Thus, the increase in nuclear Trx1 levels did not correspond to an increase in the overall reducing capacity of Trx1 in the nucleus. PMID- 12753923 TI - pH-dependent photoautotrophic growth of specific photosystem II mutants lacking lumenal extrinsic polypeptides in Synechocystis PCC 6803. AB - The removal of either the PsbU or PsbV protein has been investigated in a cyanobacterial DeltaPsbO strain and in mutants carrying deletions or substitutions in lumen-exposed domains of CP47. These experiments have demonstrated a functional interaction between the PsbU protein and photosystem II (PSII) in the absence of the PsbO subunit. The control:DeltaPsbO:DeltaPsbU strain assembled PSII centers at pH 7.5 but did not evolve oxygen; however, photoautotrophic growth was restored at pH 10.0. In addition, several CP47 mutants, lacking extrinsic proteins, were obligate photoheterotrophs at pH 7.5 but photoautotrophic at pH 10.0, whereas other strains remained photoheterotrophs at alkaline pH. PMID- 12753924 TI - The Drosophila homolog of Aut1 is essential for autophagy and development. AB - The Drosophila homolog of yeast Aut1, CG6877/Draut1, is a ubiquitously expressed cytosolic protein. Draut1 loss of function was achieved by expression of an inverted repeat transgene inducing RNA interference. The effect is temperature dependent and resembles an allelic series as described by Fortier, E. and Belote, J.M. (Genesis 26 (2000) 240-244). Draut1 loss of function larvae are unable to induce autophagy and heterophagy in fat body cells before pupariation and die during metamorphosis. To our knowledge, this is the first report of a multicellular animal lacking the function of a gene participating in the protein conjugation systems of autophagy. PMID- 12753925 TI - The X-ray structure of photosystem II reveals a novel electron transport pathway between P680, cytochrome b559 and the energy-quenching cation, ChlZ+. AB - When water oxidation by photosystem II (PSII) is impaired, an oxidized chlorophyll (Chl(Z)(+)) is formed that quenches excitation and may prevent photodamage. Both the identification of this Chl(+) and the mechanism of its oxidation and reduction are controversial. Using the available X-ray structures of PSII we calculated the efficiency of two proposed quenchers, Chl(Z)(+)(D1) and Chl(Z)(+)(D2). Of these two, only Chl(Z)(+)(D1) can quench to the degree observed experimentally. We also identify a chain of closely spaced pigments in the structure from Thermosynechococcus vulcanus that we propose to form a novel electron transport pathway between Chl(Z)(D1), beta-carotene, P680(+) and cytochrome b(559). PMID- 12753926 TI - Crystal structure of DsbDgamma reveals the mechanism of redox potential shift and substrate specificity(1). AB - The Escherichia coli transmembrane protein DsbD transfers electrons from the cytoplasm to the periplasm through a cascade of thiol-disulfide exchange reactions. In this process, the C-terminal periplasmic domain of DsbD (DsbDgamma) shuttles the reducing potential from the membrane domain (DsbDbeta) to the N terminal periplasmic domain (DsbDalpha). The crystal structure of DsbDgamma determined at 1.9 A resolution reveals that the domain has a thioredoxin fold with an extended N-terminal stretch. In comparison to thioredoxin, the DsbDgamma structure exhibits the stabilized active site conformation and the extended active site alpha2 helix that explain the domain's substrate specificity and the redox potential shift, respectively. The hypothetical model of the DsbDgamma:DsbDalpha complex based on the DsbDgamma structure and previous structural studies indicates that the conserved hydrophobic residue in the C-X-X C motif of DsbDgamma may be important in the specific recognition of DsbDalpha. PMID- 12753927 TI - Down-regulation of ARC contributes to vulnerability of hippocampal neurons to ischemia/hypoxia. AB - ARC is a caspase recruitment domain-containing molecule that plays an important role in the regulation of apoptosis. We examined ARC expression during neuronal cell death following ischemic injury in vivo and in vitro. After exposure to transient global ischemic conditions, the expression of ARC was substantially reduced in the CA1 region of hippocampus in a time-dependent manner with concomitant increase of TUNEL-positive cells. Quantitative analysis using Western blotting exhibited that most of ARC protein disappeared in the cultured hippocampal neurons exposed to hypoxia for 12 h and showing 60% cell viability. Forced expression of ARC in the primary cultures of hippocampal neurons or B103 neuronal cells significantly reduced hypoxia-induced cell death. Further, the C terminal P/E rich region of ARC was effective to attenuate hypoxic insults. These results suggest that down-regulation of ARC expression in hippocampal neurons may contribute to neuronal death induced by ischemia/hypoxia. PMID- 12753928 TI - Complementation of Escherichia coli ubiF mutation by Caenorhabditis elegans CLK 1, a product of the longevity gene of the nematode worm. AB - Caenorhabditis elegans CLK-1 was identified from long-lived mutant worms, and is believed to be involved in ubiquinone biosynthesis. The protein belongs to the eukaryotic CLK-1/Coq7p family, which is also similar to the bacterial Coq7 family, that hydroxylates demethoxyubiquinone, resulting in the formation of hydroxyubiquinone, a precursor of ubiquinone. In Escherichia coli, the corresponding reaction is catalyzed by UbiF, a member of a distinct class of hydroxylase. Although previous studies suggested that the eukaryotic CLK-1/Coq7 family is a hydroxylase of demethoxyubiquinone, there was no direct evidence to show the enzymatic activity of the eukaryotic CLK-1/Coq7 family. Here we show that the plasmid encoding C. elegans CLK-1 supported aerobic respiration on a non fermentable carbon source of E. coli ubiF mutant strain and rescued the ability to synthesize ubiquinone, suggesting that the eukaryotic CLK-1/Coq7p family could function as bacterial UbiF. PMID- 12753929 TI - Ca2+-induced oxidative stress in brain mitochondria treated with the respiratory chain inhibitor rotenone. AB - In this study we show that micromolar Ca(2+) concentrations (>10 microM) strongly stimulate the release of reactive oxygen species (ROS) in rotenone-treated isolated rat forebrain mitochondria. Ca(2+)-stimulated mitochondrial ROS release was associated with membrane lipid peroxidation and was directly correlated with the degree of complex I inhibition by rotenone. On the other hand, Ca(2+) did not increase mitochondrial ROS release in the presence of the complex I inhibitor 1 methyl-4-phenylpyridinium. Cyclosporin A had no effect on Ca(2+)-stimulated mitochondrial ROS release in the presence of rotenone, indicating that mitochondrial permeability transition is not involved in this process. We hypothesized that Ca(2+)-induced mitochondrial oxidative stress associated with partial inhibition of complex I may be an important factor in neuronal cell death observed in the neurodegenerative disorder Parkinson's disease. PMID- 12753930 TI - Plant polyphenols inhibit VacA, a toxin secreted by the gastric pathogen Helicobacter pylori. AB - VacA is a major virulence factor of the widespread stomach-dwelling bacterium Helicobacter pylori. It causes cell vacuolation and tissue damage by forming anion-selective, urea-permeable channels in plasma and endosomal membranes. We report that several flavone derivatives and other polyphenols present in vegetables and plants inhibit ion and urea conduction and cell vacuolation by VacA. Red wine and green tea, which contain many of the compounds in question, also potently inhibit the toxin. These observations suggest that polyphenols or polyphenol derivatives may be useful in the prevention or cure of H. pylori associated gastric diseases. PMID- 12753931 TI - Two variants of zebrafish p100 are expressed during embryogenesis and regulated by Nodal signaling. AB - Human p100 protein was first identified as a transcriptional coactivator of Epstein-Barr virus nuclear antigen 2, and has been shown to be a coactivator of other cellular transactivators. Its roles in development of vertebrate embryos, however, have not been reported. We have identified a zebrafish ortholog of the human p100 coactivator. The zebrafish p100 transcript is processed to two alternative variants, long and short forms, referred to as p100L and p100S, respectively. Both GFP-p100L and GFP-p100S fusion proteins are located in the cytoplasm of transfected culture cells and microinjected embryonic cells. Analysis of transcripts with Northern blots revealed the presence of p100L and lower amounts of p100S mRNAs from the one-cell stage throughout the life cycle. Whole-mount in situ hybridization shows that p100L and p100S share the same spatiotemporal expression pattern. Their zygotic expression is initially restricted to axial mesoderm precursors during gastrulation, and then spreads over other tissues during segmentation, and finally is constrained to some internal organs at day 5. We also find that Nodal signaling is essential for the zygotic expression of p100. These studies pave the way to understanding in depth the role of p100 during vertebrate embryogenesis. PMID- 12753932 TI - Origin of the scaling rule for fundamental living organisms based on thermodynamics. AB - The regular relationships between metabolic energy and body mass M of unicellular organisms, poikilotherms and homeotherms were well known as general equations. The metabolic energy rate and the life span are proportional to M(0.75) and to M(0.25), respectively. As a result, the product of the metabolic energy rate and the life time, namely, life metabolic energy, is proportional to the mass of the living organism. The origin of the scaling rules for environmental organizing systems is as follows: (1) the scaling rules for internal energy, activation energy and free energy as a function of temperature and mass of a mole of molecules. (2) The majority of species of the living organisms have the same molecules such as polysaccharides, lipids, proteins and nucleic acids in nearly same the ratio. (3) The internal energy of reactants in living organisms is equilibrium with the internal energy of water. Then, the integrated metabolic energy over the synthesizing time depends on internal energy of water and is proportional to mass M, despite the synthesizing time of the system depending on reaction rate. The proportional constant is obtained based on the thermodynamics for fundamental living organisms such as unicellular organisms and plants. Information on the environmental organizing system is also discussed. PMID- 12753933 TI - Graphic modeling of epithelial transport system: causality of dissipation. AB - The epithelial transport system is a thermodynamic system which is composed of membranes and fluid compartments. The membranes are assumed to be dissipative subsystems in which power dissipates, and fluid compartments are capacitive subsystems in which power is stored. Each subsystem can be subdivided into elementary thermodynamic processes, and can be represented by generalized capacitors, power transducers and resistors in a bond graph. In the modeling of the dissipative subsystem, the causality of the dissipative process was taken into consideration and the representation of power coupling was developed. The dissipative subsystem can be represented by a combination of coupling modules and conductors. Phenomenological equations with parameters from the model were derived. This study shows that the behavior of transport systems can be simulated using these equations. PMID- 12753934 TI - A biochemical logic gate using an enzyme and its inhibitor. Part II: The logic gate. AB - Enzyme-Based Logic Gates (ENLOGs) are key components in bio-molecular systems for information processing. This report and the previous one in this series address the characterization of two bio-molecular switching elements, namely the alpha chymotrypsin (alphaCT) derivative p-phenylazobenzoyl-alpha-chymotrypsin (PABalphaCT) and its inhibitor (proflavine), as well as their assembly into a logic gate. The experimental output of the proposed system is expressed in terms of enzymic activity and this was translated into logic output (i.e. "1" or "0") relative to a predetermined threshold value. We have found that an univalent link exists between the dominant isomers of PABalphaCT (cis or trans), the dominant form of either acridine (proflavine) or acridan and the logic output of the system. Thus, of all possible combinations, only the trans-PABalphaCT and the acridan lead to an enzymic activity that can be defined as logic output "1". The system operates under the rules of Boolean algebra and performs as an "AND" logic gate. PMID- 12753935 TI - Biological nonlocality and the mind-brain interaction problem: comments on a new empirical approach. AB - Up to now, we have been faced with an age old fundamental dilemma posed by the mind-brain interaction problem, i.e. how is it that the mind which is subjective and immaterial, can interact with the brain which is objective and material? Analysis of recent experiments appears to indicate that quantum mechanics may have a role to play in the resolution of the mind-brain interaction problem in the form of biological entanglement and nonlocality. In addition to this analysis, when coupled with ongoing and proposed experiments, may help us to simultaneously resolve related issues such as whether mental events can initiate neural events, the transference of conscious subjective experience, the measurement problem and the binding problem. PMID- 12753936 TI - On the physics of the symbol--matter problem in biological systems and the origin of life: affine Hilbert spaces model of the robustness of the internal quantum dynamics of biological systems. AB - In the present paper, some physical considerations of the biological symbol matter problem is exposed. First of all, the physical concept of quantum dynamical internal measuremental robustness is discussed. In this context, the significance of introducing affine molecular Hilbert spaces, the original (primordeal) internal quantum measurement, and the global constraining nature of time-inversion symmetry restoring, as a special restoration force, is discussed at some length. It is pointed out, as a summary, that global robustness of the internal dynamics of quantum measurements is due to two basic factors: on one hand, the global constraining nature of the chosen specific (symmetry-) restoring force, and on the other, the individual robustness of the discrete local internal measuremental interactions. The second condition is supposed to follow from a system-internalised ("objective") Bohr-type Copenhagen interpretation of quantum mechanics, corresponding, in an external context, to the Generalized Complementarity Principle of Bohr and Elsasser. It is not claimed, however, that this latter problem has been, as yet, satisfactorily settled physically. In fact, if it were, it would amount to a specifically biological quantum theory of internal measurement, which had to be rooted in the original primordeal global internal measurement, amounting to the origin of the genetic code. PMID- 12753937 TI - Modeling of bone formation and resorption mediated by parathyroid hormone: response to estrogen/PTH therapy. AB - Bone, a major reservoir of body calcium, is under the hormonal control of the parathyroid hormone (PTH). Several aspects of its growth, turnover, and mechanism, occur in the absence of gonadal hormones. Sex steroids such as estrogen, nonetheless, play an important role in bone physiology, and are extremely essential to maintain bone balance in adults. In order to provide a basis for understanding the underlying mechanisms of bone remodeling as it is mediated by PTH, we propose here a mathematical model of the process. The nonlinear system model is then utilized to study the temporal effect of PTH as well as the action of estrogen replacement therapy on bone turnover. Analysis of the model is done on the assumption, supported by reported clinical evidence, that the process is characterized by highly diversified dynamics, which warrants the use of singular perturbation arguments. The model is shown to exhibit limit cycle behavior, which can develop into chaotic dynamics for certain ranges of the system's parametric values. Effects of estrogen and PTH administrations are then investigated by extending on the core model. Analysis of the model seems to indicate that the paradoxical observation that intermittent PTH administration causes net bone deposition while continuous administration causes net bone loss, and certain other reported phenomena may be attributed to the highly diversified dynamics which characterizes this nonlinear remodeling process. PMID- 12753938 TI - The general form of 0-1 programming problem based on DNA computing. AB - DNA computing is a novel method of solving a class of intractable computational problems, in which the computing speeds up exponentially with the problem size. Up to now, many accomplishments have been made to improve its performance and increase its reliability. In this paper, we solved the general form of 0-1 programming problem with fluorescence labeling techniques based on surface chemistry by attempting to apply DNA computing to a programming problem. Our method has some significant advantages such as simple encoding, low cost, and short operating time. PMID- 12753939 TI - Pathophysiology of endometrial bleeding. AB - OBJECTIVES: To review the morphological and molecular events responsible for uterine bleeding in health and disease. METHODS: Review of pertinent literature focusing on the histology and pathophysiology of normal and abnormal uterine bleeding (AUB). RESULTS: The seat of normal menstrual bleeding is located in the upper two-thirds of the endometrial mucosa and is recognized by tissue necrosis, disruption of microvasculature, migratory leukocytes and platelet/fibrin thrombi in microvessels. The molecular events responsible for tissue and vascular breakdown are related to the release of proteolytic lysosomal enzymes of endometrial cell and inflammatory cell origin. In cases of AUB, tissue breakdown is located in the superficial layer (subsurface) of the endometrium. It is either focal (breakthrough bleeding) or diffuse (withdrawal bleeding). It is initiated by either chronic endometritis and/or microerosions or vascular fragility due to structural abnormalities of microvessels. Endometritis and microerosions occur in otherwise normal endometrium, polyps, submucosal leiomyomata, atrophy and cancer (organic causes). Primary vascular alterations are found in hyperestrogenic-type endometria, i.e. anovulatory dysfunctional uterine bleeding (DUB) and progestational-type endometrium, i.e. progestational contraceptives and combined, continuous hormonal replacement therapy (HRT) (non-organic causes). Ovulatory DUB and coagulation disorders are not appreciated histologically. These are related to impaired vasoconstriction and fibrinolysis and impaired coagulation factors, respectively. CONCLUSIONS: Histology may contribute to better understanding of the mechanisms of action that initiate, regulate and lead to AUB. Better insight may trigger in the development of therapeutic procedures that could either prevent or control vascular breakdown which results in unexpected uterine bleeding. PMID- 12753940 TI - An integrative review on current evidence of testosterone replacement therapy for the andropause. AB - OBJECTIVES: This paper examines the evidence supporting testosterone replacement in aging males. Confounding factors contributing to low testosterone levels and challenges to diagnosis of the andropause will also be considered. METHODS: A thorough review using an integrative approach citing published literature and the ongoing work of the authors. A search was performed using National Library of Medicine PubMed. Electronic and print journals available at the Texas Medical Center library were also considered. RESULTS: Information based on collective trials in older men has added to evidence for benefits and side effects of testosterone replacement inferred from studies in younger hypogonadal patients and animal models. In general, most investigators agree with short-term safety but long-term safety is unknown. Testosterone therapy in aging males improves body composition, certain domains of brain function and may also decrease cardiovascular risk in biological models. Measurable clinical effects are less apparent. Potential risks include erythrocytosis, edema, gynecomastia, and prostate stimulation. The possibility of increased risk of clinically significant prostate cancer and cardiovascular disease has been considered. CONCLUSION: The search continues for an ideal replacement androgen and larger long-term studies are needed. At this time, androgen replacement is on a case-by-case basis and prostate cancer screening should be completed prior to instituting therapy. Routine androgen replacement therapy for aging males will have significant economic implications, and is not currently recommended. PMID- 12753941 TI - Menopause after breast cancer: a survey on breast cancer survivors. AB - Due to the younger age and the ever wider use of adjuvant chemotherapy and antiestrogens, menopausal symptoms are a frequent cause of concern for breast cancer patients. OBJECTIVES: To determine the prevalence of menopausal symptoms, and to explore the attitudes toward Hormone Replacement Therapy (HRT) or other treatments and the willingness to take oestrogen in breast cancer patients. METHODS: A questionnaire-based survey on 250 breast cancer patients treated and followed-up at our department. Of them 144 (Group A) were in postmenopause and 106 (Group B) were in premenopause at time of diagnosis. RESULTS: Adjuvant therapy with tamoxifen or tamoxifen plus chemotherapy is associated with a significant worsening of menopause-related symptoms of women belonging to Group A. These women are more concerned about risk of breast cancer recurrence than about risk of osteoporosis (P=0.05) and heart disease (P=0.006). Seventy-eight percent are against the use of HRT; only 22% would consider taking HRT mainly for vasomotor symptoms relief and osteoporosis prevention. The incidence of vasomotor and dystrophic symptoms is significantly higher in women belonging to Group B treated with chemotherapy and/or hormonotherapy as compared with postmenopausal women (P<0.000 and P=0.02, respectively). Premenopausal women are more concerned about risk of breast cancer recurrence than older women (P=0.09) and at the same time are significantly more worried about the impairment of the quality of life due to adjuvant therapy (P=0.005). Younger women are more prone to consider HRT than postmenopausal women (P=0.05). Sixty-six percent are against HRT use, and 34% would consider taking HRT to alleviate vasomotor and dystrophic symptoms and to prevent osteoporosis. CONCLUSIONS: Breast cancer survivors are interested to treatments that may improve their quality of life, but fear of HRT persists among these women and their doctors, despite new evidence suggesting the low probability of detrimental effects. PMID- 12753942 TI - Effects of uncoupling protein 1 and beta3-adrenergic receptor gene polymorphisms on body size and serum lipid concentrations in Japanese women. AB - OBJECTIVES: To investigate whether the -3826 A-G point mutation of the uncoupling protein 1 (UCP1) gene and the Trp64Arg mutation of the beta3-adrenergic receptor (beta3-AR) gene are associated with increased susceptibility to weight gain and hyperlipidemia in postmenopausal women. METHODS: We genotyped 312 Japanese women for UCP1 and beta3-AR gene polymorphisms, and investigated their effects on anthropometrical parameters, serum lipid concentrations, and their changes after 4 years. RESULTS: Although body mass index (BMI), serum triglyceride, total cholesterol, and low-density lipoprotein levels were significantly higher and high-density lipoprotein levels significantly lower in postmenopausal (n=182) than in premenopausal (n=99) women, there was no significant difference in these parameters between carriers and non-carriers of the G allele in the postmenopausal women. In the premenopausal women, BMI and the 4-year change in body weight (BW) of carriers of the G allele were significantly higher than those of non-carriers of the G allele (P=0.022 and 0.048, respectively). In the postmenopausal women, the 4-year change in the level of serum triglyceride of carriers of the G allele was significantly higher (P=0.049), and the change of high-density lipoprotein was significantly lower (P=0.020) than those of non carriers of the G allele. The beta3-AR polymorphism showed no apparent affect on these parameters. CONCLUSIONS: The -3826 A-G polymorphism of the UCP1 gene is associated with an increase in BW of premenopausal women and with the 4-year changes in serum triglyceride and high-density lipoprotein levels in postmenopausal women. PMID- 12753943 TI - Does the beneficial effect of HRT on endothelial function depend on lipid changes. AB - OBJECTIVES: To determine whether improvement in endothelial function of the brachial artery observed in women treated with hormone replacement therapy (HRT) may be explained by changes in lipid profile or blood pressure, information was used obtained in a single-centre, randomised, double blind, placebo-controlled trial. METHODS: Hundred-and-five healthy postmenopausal women, aged 50-65 years, were treated with 0.625 mg conjugated equine estrogens (CEE) combined with 2.5 mg medroxyprogesterone acetate (MPA) (CEE+MPA), 2.5 mg tibolone or placebo for 3 months. At baseline and after 3 months, endothelial function was assessed using flow-mediated dilatation (FMD) and nitro glycerine-mediated dilatation (NMD). Furthermore, lipids were measured. Multivariate linear regression analysis was applied to address the research question. RESULTS: Treatment with CEE+MPA resulted in an improvement in FMD of 2.0% (95% CI: -0.1; 4.1). CEE/MPA reduced total cholesterol with 13% (95% CI: -18%; -7%), LDL-cholesterol with 23% (95% CI: -30%; -15%) and lipoprotein(a) (Lp(a)) with 14% (95% CI: -26%; -2%). The magnitude of the relation of CEE/MPA with endothelial function was attenuated to from 2.0 to 1.6% when change in Lp(a) was taken into account. Adjustments for other lipids or blood pressure did not attenuate the association. CONCLUSIONS: The improvement in endothelial function in postmenopausal women treated with CEE+MPA appears to be partially mediated by change in Lp(a), and apparently not by changes in other lipids. PMID- 12753944 TI - Vaginal dryness assessment in postmenopausal women using pH test strip. AB - OBJECTIVE: To objectively analyze vaginal dryness in postmenopausal women. METHODS: Forty healthy postmenopausal women, were divided in three groups according to the treatment they received. Group I, conjugated equine estrogens (CEE) 0.625 mg/day (n=20), group II, CEE 0.625 mg/day plus chlormadinone 1 mg/day (n=13), and group III, CEE 0.625 mg/day plus medroxyprogesterone 2.5 mg/day (n=7). Vaginal dryness intensity was analyzed using an analog visual scale, and vaginal humidity measuring the moistening in mm of a pH test strip. Both were evaluated at baseline and 3 months after the beginning of treatment. The comparison among groups and between the baseline and final results was done with Student's t-test for paired and independent samples, respectively. Pearson correlation analysis was carried out between final vaginal dryness intensity and the final moistening of the pH test strip. RESULTS: No statistically significant differences were found in age neither in somatometric variables among the groups. In the three groups, vaginal dryness intensity significantly decreased and the pH test strip moistening significantly increased. A significant negative correlation was only found in group II (-0.690 P<0.009). CONCLUSIONS: Vaginal dryness evaluation assessing the pH test strip moistening is an objective method to evaluate this symptom in postmenopausal women, and the results are independent of the hormone replacement therapy schedule. PMID- 12753945 TI - Effects of estradiol, cyproterone acetate, tibolone and raloxifene on uterus and aorta atherosclerosis in oophorectomized cholesterol-fed rabbits. AB - BACKGROUND: Different hormonal replacement regimens are used for treating climacteric complaints; however, not all of them have the same clinical profile. Cardiovascular disease (CVD) is a major health problem and tibolone, raloxifene, estradiol (alone or with cyproterone acetate) have been added to cholesterol-fed rabbits to study atherosclerosis. METHODS: A total of 48 cholesterol-fed New Zealand white rabbits were studied for 4 months. Forty rabbits underwent bilateral ovariectomy and the other eight were sham operated (group S). The ovariectomized rabbits were allocated to five groups of eight animals each receiving tibolone (Group T, 6 mg/day), raloxifene (R, 35 mg/day), estradiol valerate (E, 3 mg/day), estradiol valerate plus cyproterone acetate (EC, 3+0.5 mg/day, respectively), and no treatment for the control group (C). The sham group received no treatment too. RESULTS: After 4 months the percentage of the extent of atherosclerosis in the aorta was 30.4% in C group, 24.5% in S group, 10.2% in T group, 30.3% in R group, 17.9% in E group and 28.1% in EC group (P<0.05 T vs. C, R, EC). The aortic cholesterol content compared with aortic weight was 8.55 microg/mg in C group, 11.97 microg/mg in S group, 1.86 microg/mg in T group, 3.82 microg/mg in R group, 2.86 microg/mg in E group and 5.24 microg/mg in EC group (P<0.05 T vs. EC, C, S; R vs. C, S; E vs. C, S). Uterine weights in grams were: 1.89 (C group), 2.24 (S), 7.38 (T), 1.94 (R), 9.92 (E), and 5.94 (EC); P<0.05 (C, S, R, vs. T, E, EC; T vs. E; EC vs. T, E). CONCLUSION: Our study showed a decrease in the extent of aortic atherosclerosis in oophorectomized cholesterol fed rabbits treated with tibolone or estradiol, and a decrease in aortic cholesterol content in rabbits treated with tibolone, raloxifene and estradiol. However, rabbits treated with tibolone showed an increased uterine weight, which is contrary to that observed in humans. PMID- 12753946 TI - Relationship between PTH, sex steroid and bone turnover marker measurements and bone density in recently postmenopausal women. AB - OBJECTIVE: It is conceivable that, since menopause accelerates the continuous bone loss determined by age, a specific configuration of bone mass determinants during the first postmenopausal years occurs. METHODS: To establish their value as indicators of bone mass in women with recent natural menopause, we assessed relationships between bone mineral density (BMD) and age, menopausal age, body mass index (BMI), PTH, sex steroid hormones (estradiol and testosterone), and several markers of bone turnover in urine (N-telopeptide and calcium/creatinine ratio) or serum (osteocalcin (OC), total alkaline phosphatase (ALP), total and ionic calcium (iCa), phosphate (P) and magnesium (Mg)) for a group of 118 women (mean of three measurements per subject) attending a third-level menopause unit. Multivariate analysis was used in addition to Pearson's correlation to detect relationships between variables. RESULTS: Several significant associations were detected between variables under Pearson's correlation analysis, but only a few were confirmed under multivariate analysis. Thus, among the clinical traits, age was the main predictor of BMD for femoral neck (P<0.05). Estradiol (E(2)) was the only parameter that attained significance as a predictor for lumbar spine BMD (P<0.05), whereas PTH and NTx levels emerged as predictors of BMD for femoral neck (P<0.05). CONCLUSION: In this group of recently postmenopausal women, hormonal status, as defined by E(2) and PTH, and a resorption marker (NTx), revealed, together with age, as the only significant predictors of BMD. PMID- 12753947 TI - Biology of the Cell-new team, new topics, new look. PMID- 12753948 TI - Nitric oxide and S-nitrosylation: excitotoxic and cell signaling mechanism. AB - This review focuses on the important physiological messenger, nitric oxide (NO), and its role in N-methyl-D-aspartate (NMDA) excitotoxicity. NO has been shown to be a key mediator of voltage-gated Ca(+2) transmembrane proteins. It remains unclear whether NO is implicated during hypoxia, or ischemic/reperfusion injuries as a neuroprotective or neurodegenerative factor. Excitotoxicity results from the excessive stimulation of excitatory glutamate receptors within the CNS. This review maintains that the feed-forward pathway precipitated by oxidative stress is the discriminating factor in the neuroprotective or neurodegenerative actions of NO. PMID- 12753949 TI - Influence of age, castration, and testosterone on T cell subsets in healthy and leukemia grafted mice. AB - The distribution of T cell subsets in pubertal (2 months) and post-pubertal (10 months) mice showed a significant decrease in the percentage of CD4+ splenocytes and peripheral blood lymphocytes (PBL) with age, unlike the percentage of CD8+ cells in PBL, which remained unchanged. The change in the distribution of T cell subsets in the spleen and blood occurred in 2 months old castrated mice, as in 10 months old animals. P388 tumor grew better in post-pubertal and in castrated mice than in young mice. The intact mice survived longer than the castrated ones. The relative number of CD4+, CD8+ and CD2+ splenocytes was lower in transplanted intact mice than that in controls. The CD8+ and CD2+ subsets in the blood of 2 months transplanted mice were higher than those in controls, whereas in PBL, in 10 months old and castrated mice, the T lymphocyte subsets remain unchanged. Depo testosterone (DT) injection strongly reduced weight and tumor growth in all the intact and castrated animals. A significant correlation is observed between the tumor weight and testosterone level in the plasma of the 2 months old DT treated mice. Moreover, DT injection induced a significant increase in the percentage of blood CD8+ cells in all the batches. These data indicate that physiologically, androgens affect the age-related distribution of lymphocyte T subsets and suggest that they slow down tumor growth, besides causing a direct effect, through an immunological process. PMID- 12753950 TI - Human papillomaviruses type 16+ and 18+ cervical carcinoma cells are sensitive to staurosporine-mediated apoptosis. AB - We have recently shown that staurosporine (ST) can trigger apoptosis of CaSki and HeLa cervical tumor cells from G2/M checkpoint, though the mechanism remains elusive. In this study, we reported that ST induced the inhibition of E6 and E7 viral oncogene and MDM2 expression, while it led to increased levels of p53, which was transiently located to mitochondria. Additionally, the proteins of the p53-regulated genes, p21(WAF1) and Bax, were increased with a similar time, while Bcl-2 and Bcl-X(L) expression was lowered. Upon ST treatment, the cytochrome c was released into the cytosol and, then, activation of caspases-9 and -3 led to Poly(ADP)RibosePolymerase (PARP) cleavage. Finally, characteristic morphological signs confirmed the apoptosis execution. Thus, taken together, all these observations suggest that apoptosis can be reactivated in HPV-positive human carcinoma cells and highlight that ST could be used as a potently chemotherapy agent to enhance the sensitivity of tumor cells to apoptosis. PMID- 12753951 TI - Ultrastructure of post-fertilization development in the red alga Scinaia articulata (Galaxauraceae, Nemaliales, Rhodophyta). AB - The ultrastructure of the carposporophyte and carposporogenesis is described for the red alga Scinaia articulata Setch. After fertilization, the trichogyne disappears, and the pericarp develops to form a thick protective tissue that surrounds the carposporophyte. The hypogynous cell cuts off both one-celled and two-celled sterile branches. Patches of chromatin are frequently observed in evaginations of the nuclear envelope, which appear to produce vesicles in the cytoplasm of the cell of the sterile branch. Large gonimoblast lobes extend from the carpogonium and cleave to form gonimoblast initials. Subsequently, a fusion cell is formed from fusions of the carpogonium, the hypogynous cell and the basal cell of the carpogonial branch. The mature carposporophyte comprises the fusion cell that is connected to the sterile branch cells, gonimoblast cells and carpospores and is surrounded by extensive mucilage. Young carpospores possess a large nucleus and proplastids with a peripheral thylakoid, but they have few dictyosomes and starch granules and are indistinguishable from gonimoblast cells. Subsequently, dictyosomes are formed, which produce vesicles with an electron dense granule, which indicates an initiation of wall deposition. Thylakoid formation coincides with incipient starch granule deposition. The nuclear envelope produces fibrous vacuoles and concentric membrane bodies. Carpospores are interconnected by pit connections with two cap layers. Dictyosome activity increases, resulting in the production of vesicles, which either continue to deposit wall material or coalesce to form fibrous vacuoles. The final stage of carposporogenesis is characterized by the massive production of cored vesicles from curved dictyosomes. Mature carpospores are uninucleate and contain fully developed chloroplasts, numerous cored vesicles, numerous starch granules and fibrous vacuoles. The mature carpospore is surrounded by a wall layer and a separating layer, but a carposporangial wall is lacking. PMID- 12753952 TI - Structural inheritance in Paramecium: ultrastructural evidence for basal body and associated rootlets polarity transmission through binary fission. AB - One main difference between basal bodies and centrioles resides in the expression of their polarity: centrioles display a structural nine-fold radial symmetry, whereas basal bodies express a circumferential polarity, thanks to their asymmetric set of rootlets. The origin of this polarity during organelle duplication still remains under debate: is it intrinsic to the nine-fold structure itself (i.e. the nine microtubular triplets are not equivalent) or imposed by its immediate environment at time of assembly? We have reinvestigated this problem using the Ciliate Paramecium, in which the pattern of basal body duplication is well known. In this cell, all basal bodies produced within ciliary rows appear immediately anterior to parental ones. Observations on cells fixed with the tannic acid protocol suggest that, to be competent for basal body assembly, parental basal bodies have to be individually associated with a complete set of rootlets (monokinetid structure). During pro-basal body assembly, full microtubular triplets were detected according to a random circumferential sequence; during the whole process, the new basal body and its associated rootlets maintained structural relations with the parental monokinetid structure by way of specific links. These results strongly suggest that basal body and associated rootlets (kinetid) polarity is driven by its immediate environment and provide a basis for the structural heredity property observed by Sonneborn some decades ago. PMID- 12753953 TI - Molecular characterization of a defensin in the IZD-MB-0503 cell line derived from immunocytes of the insect Mamestra brassicae (Lepidoptera). AB - The induction of anti-microbial peptides against Gram positive and negative bacteria in the IZD-MB-0503 cell line from the lepidopteran Mamestra brassicae is demonstrated, while no anti-fungal activity is detected. The identification of a defensin-like molecule active against Gram positive bacteria is described for the first time in Lepidoptera. This molecule shows between 43% and 59% homology with group A defensins from other dipteran and hymenopteran species. PMID- 12753954 TI - Distractibility during selection-for-action: differential deficits in Huntington's disease and following frontal lobe damage. AB - Selective attention can be measured through analysis of errors and reaction time (RT) for trials in which targets are presented alone compared with trials in which targets and distractors are presented. This study investigated selective attention using a reaching task, in which subjects made rapid reaches to targets. Thirty-seven patients with lesions of the prefrontal cortex (PFC) were compared with 19 healthy age- and IQ-matched volunteers and 18 patients with early-stage Huntington's disease (HD), a neurodegenerative disease primarily affecting the basal ganglia. It was hypothesised that, if fronto-striatal circuits as a whole support selection-for-action, then the pattern of behavioural performance of both patient groups would be similar. Alternatively, if the functional roles of PFC and basal ganglia in selection-for-action are dissociable, then two different patterns would emerge. It was found that that both HD and frontal groups were significantly more distractible than controls for RT, but they had a different pattern of errors. Frontal patients made significantly more touches of the distractor location itself than did controls, while this was not the case for HD. It is argued that a reactive-inhibition mechanism, required in the circumstance of strong distractor activation, is affected by frontal damage, while a lateral inhibition mechanism, invoked during the recruitment of selective attention, is affected in HD. Additionally, there were significant correlations between the degree of distractibility for RT and the extent of lateral PFC damage, and between cue-generated preparation and lateral PFC damage, thus highlighting the critical importance of lateral, rather than orbital or medial, PFC for attention to action. PMID- 12753955 TI - A combined neuropsychological and neuroimaging study of topographical and non verbal memory in semantic dementia. AB - A combined neuropsychological and neuroimaging investigation was carried out on a patient (O.I.) with semantic dementia who had asymmetrical temporal lobe atrophy, greater on the left. His performance on tests of verbal memory was gravely impaired. Similarly, his visual memory as indexed by recognition of unfamiliar faces was impaired. By contrast, his recognition memory for topographical memoranda (e.g. buildings, landscapes) and ability to find his way around was preserved. In order to identify the neural substrates supporting the preserved recognition of static topographical memoranda, O.I. was scanned using positron emission tomography (PET) during the encoding and recognition of building and landscape stimuli. In common with control subjects, during encoding O.I. activated parahippocampal cortex bilaterally, along with bilateral temporo parietal, retrosplenial and left frontal cortices. During recognition, both patient and controls activated right parahippocampal, right superior parietal and right frontal cortices. Notably, control subjects, but not O.I., also activated at encoding the precuneus and at recognition the retrosplenial cortex. This allows the conclusion that these two areas while involved may not be necessary for topographical memory. Interestingly, the patient also activated regions that were not evident in control subjects both during encoding and recognition. These additional areas of activation may be necessary in a compensatory role. Overall, these data represent the first reported assessment of the functional integrity of degenerating brain tissue and its contribution to preserved topographical memory. The combination of the neuropsychological and neuroimaging approaches may provide insights into the functional-anatomy of memory while having clinical utility for the assessment of residual brain tissue. PMID- 12753956 TI - Involvement of the mediodorsal thalamic nucleus in mediating recollection and familiarity. AB - The mediodorsal (MD) thalamic nucleus is thought to play an important role in memory processes. Distinct hippocampal-thalamic-prefrontal connections have been described as the potential neural substrate for both, recollection and familiarity. The aim of this study was to investigate whether the MD is part of the circuits underlying these two memory components. We assessed the effects of ischemic thalamic lesions with or without MD involvement on performance in a word list discrimination task and standard tests of memory and executive function. Estimates of recollection and familiarity were derived using the dual-process signal-detection model (DPSD). The results revealed impairments in both, recollection and familiarity, after unilateral thalamic damage, with recollection being more affected than familiarity. There were no significant differences in the memory performance of patients with MD lesions compared to patients with ventrolateral-thalamic lesions except for familiarity estimates, which were lower for the latter group. Lesions involving the MD led to recollection deficits, although inspection of individual cases suggested a decrease in both memory components after damage in the medial part of this nucleus. Executive dysfunction was associated with lateral MD lesions and also ventrolateral-thalamic damage. The findings suggest that MD contributes to recollection, with some preliminary evidence of a contribution of the medial MD to familiarity. The small sample size does, however, not yet allow any clear conclusions in this regard. Since damage in the ventrolateral thalamus leads to memory and executive dysfunction, further research is needed to elucidate the role of this thalamic region in cognition. PMID- 12753957 TI - Patient assessment based on a theory of visual attention (TVA): subtle deficits after a right frontal-subcortical lesion. AB - We report on a patient who complained of reduced awareness in the left visual field, but showed no visual neglect or extinction in clinical testing. By MR scanning, the brain damage was localized to the right basal ganglia, also involving structures in right frontal cortex. Using psychophysical testing and mathematical modeling based on Bundesen's theory of visual attention [TVA; Psychol. Rev. 97 (1990) 523], the patient's subjective experience of attentional disturbance was confirmed, and the deficit was specified into several components. At very short exposure durations, two effects were shown. The detection threshold was elevated, particularly in the left visual field, and stimuli in this side were given less attentional weight. In addition, the capacity of visual short term memory (VSTM) was markedly reduced in both visual fields. The robustness of the test results was evaluated by bootstrap analysis. The study demonstrates the sensitivity and specificity gained by combining psychophysical testing with TVA modeling in the analysis of visual attention disorders. Extending the results of a pioneer study of parietal neglect patients by Duncan et al. [J. Exp. Psychol.: Gen. 128 (1999) 450], this study demonstrates the strength of the method in a single case, with a lesion outside parietal cortex, and only minor clinical symptoms. PMID- 12753958 TI - Grammatical processing of nouns and verbs in left frontal cortex? AB - We report the case of a brain-damaged subject R.C. who is more impaired at producing grammatical forms of words and pseudo-words used as verbs (he judges, he wugs) than of the same words used as nouns (the judges, the wugs). This pattern of performance constitutes the first clear demonstration that grammatical knowledge about verbs can be selectively impaired following brain damage. A comparison of R.C.'s behavioral and neurological profile with that of a patient who shows similar difficulties with nouns suggests that nouns and verbs are processed by separate neural systems with components in the left frontal lobe. PMID- 12753959 TI - Linguistic theory and neuroimaging evidence: an fMRI study of Broca's area in lexical semantics. AB - There has been a long debate on the functional characterization of left inferior frontal cortex, including proposals regarding syntactic and lexico-semantic involvement. We studied nine right-handed adults, using functional magnetic resonance imaging (fMRI) during performance on a semantic decision task in which subjects had to determine whether noun-verb pairs were semantically associated. In comparison with a visuoperceptual control task, activation clusters were seen in left inferior frontal and middle temporal regions, as well as the bilateral superior frontal gyrus. In agreement with previous studies, our findings suggest that Broca's area is involved in semantic processing. Findings of lexico-semantic as well as syntactic processing in the inferior frontal lobe may be accounted for in terms of working memory demands. PMID- 12753960 TI - Material-dependent and material-independent selection processes in the frontal and parietal lobes: an event-related fMRI investigation of response competition. AB - The present study used the flanker task [Percept. Psychophys. 16 (1974) 143] to identify neural structures that support response selection processes, and to determine which of these structures respond differently depending on the type of stimulus material associated with the response. Participants performed two versions of the flanker task while undergoing event-related functional magnetic resonance imaging (fMRI). Both versions of the task required participants to respond to a central stimulus regardless of the responses associated with simultaneously presented flanking stimuli, but one used colored circle stimuli and the other used letter stimuli. Competition-related activation was identified by comparing Incongruent trials, in which the flanker stimuli indicated a different response than the central stimulus, to Neutral stimuli, in which the flanker stimuli indicated no response. A region within the right inferior frontal gyrus exhibited significantly more competition-related activation for the color stimuli, whereas regions within the middle frontal gyri of both hemispheres exhibited more competition-related activation for the letter stimuli. The border of the right middle frontal and inferior frontal gyri and the anterior cingulate cortex (ACC) were significantly activated by competition for both types of stimulus materials. Posterior foci demonstrated a similar pattern: left inferior parietal cortex showed greater competition-related activation for the letters, whereas right parietal cortex was significantly activated by competition for both materials. These findings indicate that the resolution of response competition invokes both material-dependent and material-independent processes. PMID- 12753961 TI - Phineas gauged: decision-making and the human prefrontal cortex. AB - Poor social judgment and decision-making abilities have often been attributed to people who have suffered injury to the ventromedial prefrontal cortex (VMPFC). However, few laboratory tests of decision-making have been conducted on these patients. The exception to this is the Iowa Gambling Task which has often, but not always, demonstrated differential performance between patients and controls. Results from patients with prefrontal cortex lesions on a novel test of decision making are presented. Participants explored and chose from pairs of gambles that differed in their underlying distributions, primarily in the variance of their respective outcomes. In accordance with many findings from the behavioral decision-making literature, both young normal participants and older patient controls demonstrated a marked avoidance of risk and selected largely from secure, low variance gambles. In contrast, patients with ventromedial lesions were divided into two clear sub-groups. One group behaved similarly to normals, showing a risk-averse strategy. The other group displayed a distinctive risk seeking behavior pattern, choosing predominantly from the high-variance, high risk decks. This research demonstrates some of the advantages of using methods and theories from traditional decision-making research to study the behavior of patients, as well as the benefits of examining individual participants, and provides new insights into the nature of the decision-making deficit in patients with ventromedial prefrontal cortex lesions. PMID- 12753962 TI - An investigation of errorless learning in memory-impaired patients: improving the technique and clarifying theory. AB - In rehabilitating individuals who demonstrate severe memory impairment, errorless learning techniques have proven particularly effective. Prevention of errors during acquisition of information leads to better memory than does learning under errorful conditions. This paper presents results of a study investigating errorless learning in three patient groups: those demonstrating mild, moderate, and severe memory impairments. The first goal of the study was to trial a new version of errorless learning, one encouraging more active participation in learning by patients via the use of elaboration and self-generation. This technique led to significantly better memory performance than seen under standard errorless conditions. This finding highlights the value of encouraging active and meaningful involvement by patients in errorless learning, to build upon the benefits flowing from error prevention. A second goal of the study was to clarify the mechanisms underlying errorless learning. Memory performance under errorless and errorful conditions was compared within and across each group of patients, to facilitate theoretical insight into the memory processes underlying performance. The pattern of results observed was equivocal. The data most strongly supported the hypothesis that the benefits seen under errorless learning reflect the operation of residual explicit memory processes, however a concurrent role for implicit memory processes was not ruled out. PMID- 12753963 TI - Reasoning and working memory: common and distinct neuronal processes. AB - The neuronal processes underlying reasoning and the related working memory subsystems were examined with functional magnetic resonance imaging (fMRI). Twelve volunteers solved relational reasoning problems which either supported a single (determinate) or several alternative solutions (indeterminate). In a second condition, participants maintained the identical premises of these problems in working memory without making inferences. Although problems were presented in auditory format, activity was detected for both reasoning and maintenance in a network comprising bilaterally the secondary visual cortex, the posterior cingulate cortex, and the medial anterior frontal cortex. In direct comparisons, reasoning was associated with stronger dorsolateral and medial prefrontal activation than maintenance, whereas maintenance led to stronger lateral parietal activation than reasoning. Participants' visuo-spatial abilities ("Block Design" score) covaried positively with behavioral performance and negatively with activity of the precuneus for reasoning, but not for maintenance. These results support the notion that relational reasoning is based on visuo spatial mental models, and they help to distinguish the neuronal processes related to reasoning itself versus to the maintenance of problem information in working memory. PMID- 12753964 TI - Preserved use of spatial cues for sound segregation in a case of spatial deafness. AB - Auditory spatial cues contribute to sound localisation and to sound object segregation. We have investigated these capacities in a patient (NM) who complained having difficulties to localise sounds in everyday life after a right temporo-parieto-frontal ischemic lesion. Two groups of tasks were used, in which spatial dimension was simulated by interaural time differences (ITD): (i) active localisation of stationary or moving sound targets, and (ii) sound segregation on the basis of spatial cues. This latter included a spatial release from masking paradigm and two ITD diotic tasks. NM failed to localise stationary and moving sounds: she perceived all the stimuli at the centre of the head, and could not differentiate stationary from moving targets. In contrast, NM was able to use ITD cues to segregate simultaneous sound sources in the spatial-release-from-masking paradigm and in ITD diotic tasks.These results suggest that sound localisation and sound object segregation based on spatial cues do not rely on the same mechanisms. PMID- 12753965 TI - Hemispatial neglect: its effects on visual perception and visually guided grasping. AB - Hemispatial neglect is a neurological disorder characterized by a failure to represent information appearing in the hemispace contralateral to a brain lesion. In addition to the perceptual consequences of hemispatial neglect, several authors have reported that hemispatial neglect impairs visually guided movements. Others have reported that the extent of the impairment depends on the type of visually guided task. Finally, in some cases, neglect has been shown to impair visual perception without affecting visuomotor control in relation to the very same stimuli. While neglect patients may be able to successfully pick up an object they have difficulty perceiving in its entirety, it does not mean that they are picking up the object in the same way that a neurologically intact individual would. In the current study, patients with hemispatial neglect were presented with irregularly shaped objects, directly in front of them, that lacked clear symmetry and required an analysis of their entire contour in order to calculate stable grasp points. In a perceptual discrimination task, the neglect patients had difficulty distinguishing one object from another on the basis of their shape. In a grasping task, the neglect patients showed more variance in the position of their grasp on the target objects than their control subjects, with an overall shift to the relative right side of the presented objects. The perceptual and visuomotor deficits seen in patients with hemispatial neglect deficits may be the result of an inability to form good structural representations of the entire object for use in visual perception and visuomotor control. PMID- 12753966 TI - Modulation of motor cortex excitability in the left hemisphere during action observation: a single- and paired-pulse transcranial magnetic stimulation study of self- and non-self-action observation. AB - Motor system excitability was tested by transcranial magnetic stimulation (TMS), and F-wave and H-reflex evaluation in different action observation tasks. Our aim was to investigate the effects produced by self- versus non-self-hand movement observation (MO). No significant differences were found between the self- and non self-conditions. Movement observation significantly modulated motor cortex excitability, producing an increase in the amplitude of motor evoked potentials (MEPs) with a single magnetic pulse and a reduction in intracortical inhibition (ICI) with paired-pulse stimulation. No significant changes were found in motor cortex excitability during the observation of geometric objects. Motor imagery produced similar effects to those of action observation; no significant differences in modulation of motor system excitability between motor imagery and action observation were found in those muscles involved in actual motor execution. No significant effect on spinal excitability was found in any of the test conditions. PMID- 12753968 TI - Primary metabolism and nutrient assimilation: achieving a balanced diet. PMID- 12753969 TI - New complexities in the synthesis of sucrose. AB - Sucrose is universal in plants and fulfils many roles: transport sugar, storage reserve, compatible solute and signal compound. Consequently, sucrose synthesis is highly regulated, with much of the control operating at the first step in the committed pathway, which is catalysed by sucrose-phosphate synthase (SPS). The discovery of at least three SPS gene families in plants has added a further layer of complexity to an already complicated picture involving transcriptional, allosteric and post-translational control of this enzyme's activity. After years of neglect, the gene encoding the last enzyme in the pathway, sucrose-phosphatase (SPP), has finally been cloned, revealing that SPS contains an SPP-like domain at the carboxy-terminus, to which SPP might bind. This has reinvigorated the search for an SPS-SPP complex, and has hinted at further complexities to be unravelled in the control of sucrose synthesis in plants. PMID- 12753971 TI - Fructans: beneficial for plants and humans. AB - The recent cloning of genes encoding fructosyltransferases and fructan exohydrolases has been a major breakthrough in fructan research. Now, fructan metabolism and fructosyltransferase enzymes can be studied at the molecular level. In addition, fructan synthesis and breakdown can be adapted in such a way that tailor-made fructans are produced in plants for use as healthy food ingredients. PMID- 12753970 TI - Starch synthesis in the cereal endosperm. AB - The pathway of starch synthesis in the cereal endosperm is unique, and requires enzyme isoforms that are not present in other cereal tissues or non-cereal plants. Recent information on the functions of individual enzyme isoforms has provided insight into how the linear chains and branch linkages in cereal starch are synthesized and distributed. Genetic analyses have led to the formulation of models for the roles of de-branching enzymes in cereal starch production, and reveal pleiotropic effects that suggest that certain enzymes may be physically associated. For the first time, tools for global analyses of starch biosynthesis are available for cereal crops, and are heralded by the draft sequence of the rice genome. PMID- 12753972 TI - Trehalose metabolism: a regulatory role for trehalose-6-phosphate? AB - Trehalose is a disaccharide that was initially thought to be rare in plants but now appears to be ubiquitous. A recent study has established that the initial step in trehalose synthesis is essential in Arabidopsis. Evidence is emerging that the precursor of trehalose (trehalose-6-phosphate) is an important regulatory molecule. In yeast, trehalose-6-phosphate regulates sugar influx into glycolysis. In plants, trehalose-6-phosphate also appears to regulate sugar metabolism, but the underlying mechanism is unresolved and may be substantially different from that in yeast. PMID- 12753973 TI - The oxidative pentose phosphate pathway: structure and organisation. AB - The oxidative pentose phosphate pathway is a major source of reducing power and metabolic intermediates for biosynthetic processes. Some, if not all, of the enzymes of the pathway are found in both the cytosol and plastids, although the precise distribution of their activities varies. The apparent absence of sections of the pathway from the cytosol potentially complicates metabolism. These complications are partly offset, however, by exchange of intermediates between the cytosol and the plastids through the activities of a family of plastid phosphate translocators. Molecular analysis is confirming the widespread presence of multiple genes encoding each of the enzymes of the oxidative pentose phosphate pathway. Differential expression of these isozymes may ensure that the kinetic properties of the activity that catalyses a specific reaction match the metabolic requirements of a particular tissue. This hypothesis can be tested thanks to recent developments in the application of 13C-steady-state labelling strategies. These strategies make it possible to quantify flux through metabolic networks and to discriminate between pathways of carbohydrate oxidation in the cytosol and plastids. PMID- 12753975 TI - Don't shoot the (second) messenger: endomembrane transporters and binding proteins modulate cytosolic Ca2+ levels. AB - Ca(2+) signal transduction requires the meticulous regulation of cytosolic Ca(2+) levels. Endomembrane Ca(2+) transporters and binding proteins are important components in partitioning these Ca(2+) signals to mediate cellular activity. Recently, many of these proteins have been characterized and mutant analysis suggests that these transporters form a network. Future attempts to manipulate plant Ca(2+) signaling must address all aspects of this complex. PMID- 12753974 TI - Response of plant metabolism to too little oxygen. AB - Oxygen can fall to low concentrations within plant tissues, either because of environmental factors that decrease the external oxygen concentration or because the movement of oxygen through the plant tissues cannot keep pace with the rate of oxygen consumption. Recent studies document that plants can decrease their oxygen consumption in response to low oxygen concentrations to avoid internal anoxia. This adaptive response involves a restriction of respiration and a concomitant decrease in ATP consumption that results from the inhibition of a wide range of biosynthetic processes. The inhibition of respiration is rapid and occurs at oxygen concentrations well above the K(m)(oxygen) of cytochrome oxidase, indicating that an oxygen-sensing system triggers a coordinated inhibition of ATP formation and consumption. In addition to this, low oxygen concentrations lead to the induction of a plant-specific and energy-conserving pathway of sucrose degradation, which decreases oxygen consumption and improves plant performance. Low oxygen concentrations also lead to long-term morphological adaptations, which allow respiration per volume tissue to be decreased and oxygen entry to be increased. Recently, advances have been made in elucidating possible oxygen-sensing systems and regulatory components that are involved in these responses. PMID- 12753976 TI - Genes for magnesium transport. AB - We know very little about the regulation of magnesium uptake and the control of magnesium homeostasis. After years of relative neglect, however, rapid progress is now being made in understanding the molecular biology of magnesium transport in eukaryotes. Several new gene families have been implicated, and tools are in place for the dissection of the biochemical and biological roles played by the encoded proteins. PMID- 12753977 TI - Got silicon? The non-essential beneficial plant nutrient. AB - Research on a possible nutritional role for the element silicon has been hampered by the diverse beneficial effects that it has on monocots and dicots, and the subsequent difficulties in focusing studies on a single genetic model system. Although deemed a non-essential nutrient for the majority of plants, the benefits of silicon include increasing pest and pathogen resistance, drought and heavy metal tolerance, and the quality and yield of agricultural crops. Although the pathways and molecular mechanisms by which silicon is absorbed and deposited in plants are still unclear, recent progress has been achieved through the use of rice mutants that are deficient in silicon uptake. Additionally, the application of electron-energy-loss spectroscopy (EELS) allows one to determine the composition of silica deposits conclusively. Thereby shedding light upon the role of silicon in heavy metal tolerance. With the complete sequence of the genomes for a dicot (Arabidopsis) and a monocot (rice) available for large-scale genetic analysis, the future bodes well for a more complete understanding of the biological role of silicon and its mode of transport into and through plants. PMID- 12753978 TI - Plants, selenium and human health. AB - Selenium is an essential nutrient for animals, microorganisms and some other eukaryotes. Although selenium has not been demonstrated to be essential in vascular plants, the ability of some plants to accumulate and transform selenium into bioactive compounds has important implications for human nutrition and health, and for the environment. Selenium-accumulating plants provide unique tools to help us understand selenium metabolism. They are also a source of genetic material that can be used to alter selenium metabolism and tolerance to help develop food crops that have enhanced levels of anticarcinogenic selenium compounds, as well as plants that are ideally suited for the phytoremediation of selenium-contaminated soils. PMID- 12753979 TI - The role of nutrient availability in regulating root architecture. AB - The ability of plants to respond appropriately to nutrient availability is of fundamental importance for their adaptation to the environment. Nutrients such as nitrate, phosphate, sulfate and iron act as signals that can be perceived. These signals trigger molecular mechanisms that modify cell division and cell differentiation processes within the root and have a profound impact on root system architecture. Important developmental processes, such as root-hair formation, primary root growth and lateral root formation, are particularly sensitive to changes in the internal and external concentration of nutrients. The responses of root architecture to nutrients can be modified by plant growth regulators, such as auxins, cytokinins and ethylene, suggesting that the nutritional control of root development may be mediated by changes in hormone synthesis, transport or sensitivity. Recent information points to the existence of nutrient-specific signal transduction pathways that interpret the external and internal concentrations of nutrients to modify root development. Progress in this field has led to the cloning of regulatory genes that play pivotal roles in nutrient-induced changes to root development. PMID- 12753980 TI - Insights into metabolism obtained from microarray analysis. AB - The regulation of plant metabolic processes in response to environmental and developmental signals is a complex interaction between optimization of enzyme activity and transcriptional regulation of gene expression. Through painstaking efforts over more than 50 years, many metabolic pathways in plants have been characterized and their regulation investigated, often in detail. Widely available cDNA and oligonucleotide arrays can now be used to analyze the expression profiles of a large number of genes in parallel. Coupling these expression profiles to detailed analyzes of metabolite changes will allow deep insight into the cellular mechanisms of metabolic adaptation to a wide variety of growth conditions. PMID- 12753982 TI - Tinnitus loudness matchings in relation to annoyance and grading of severity. AB - OBJECTIVE: To measure and compare different parameters of tinnitus loudness obtained via matching and to investigate their associations with self-reported severity of tinnitus. METHODS: From the audiology department 18 patients with unilateral tinnitus of sensorineural origin were selected. They underwent pure tone audiometry, and in a matching paradigm their tinnitus pitch was estimated. Loudness was assessed at tinnitus frequency and at the frequency of best hearing and both were expressed in sensation level (SL) and hearing level (HL). Moreover, minimal masking level (MML) with white noise was assessed as well as thresholds for the tinnitus tone and a comparison tone in the contralateral ear when tinnitus was masked. RESULTS: Annoyance and grading of severity were associated with measures of loudness expressed in HL, MML, and pure-tone audiometry. Loudness in SL was not associated with self-reported severity. Tinnitus loudness, matched at the frequency of best hearing instead of the tinnitus frequency, appears to be less dependent on HL, while still being associated with distress and grading of severity. CONCLUSION: Severity of tinnitus is related to hearing thresholds, and loudness in SL provides little clinically useful information. PMID- 12753981 TI - Effect of single-drug treatment on idiopathic sudden sensorineural hearing loss. AB - OBJECTIVES: In order to evaluate the effect of a medical administration for the sudden deafness patients, single-drug treatment for idiopathic sudden sensorineural hearing loss (ISSHL) was assessed at multi-centers participating in the Acute Severe Hearing Loss Study Group sponsored by the Ministry of Health, Labor and Welfare of Japan. METHODS: The subjects consisted of ISSHL patients who were (1) 20 years of age or older, (2) diagnosed within 2 weeks after the onset of hearing loss, (3) showing a mean hearing level of 40-90 dB at five frequencies from 250 to 4000 Hz, (4) previously untreated, and (5) with normal for age in hearing of the opposite ear. The drugs used in this study were ATP, alprostadil, hydrocortisone and amidotrizoate, which were administered intravenously, and beraprost sodium and betamethasone, which were given orally. Two drugs were assigned to each center, one of which was selected according to the code hidden in envelopes and administered for 1 week. The treatment after the single-drug administration was conducted at the discretion of each center. The hearing gain and recovery rate at 1 week after the initiation of single-drug treatment and at 1 month or over when the hearing level was fixed, were evaluated based on the criteria for hearing recovery prepared by the Acute Severe Hearing Loss Study Group. RESULTS: There was no statistically significant difference in the recovery rate among drugs either at 1 week after the initiation of single-drug treatment or at the time of fixed hearing level. At the time when the hearing level was fixed, a statistically significant difference in the complete recovery rate was detected only between amidotrizoate and beraprost sodium. CONCLUSION: From these results, we could not find any specific drugs recommended for ISSNHL. In evaluating the effect of the drugs, however, several problems in the clinical trial for ISSHL should be considered. PMID- 12753983 TI - CT imaging of the patulous eustachian tube--comparison between sitting and recumbent positions. AB - OBJECTIVE: The objective was to evaluate the three-dimensional anatomy of the eustachian tube (ET) and its surrounding tissues of patulous ET patients in the sitting positions by the horizontal computed tomography (CT) system with the multiplanar reconstruction (MPR) technique, and to compare the findings with those obtained in the recumbent position. METHODS: Two patients with severe bilateral patulous ET were investigated. After performing temporal bone CT examination both in the sitting and recumbent position, MPR images of the ET were reconstructed and measurement was made on a workstation. RESULTS: The ET lumen was identified at most of the portion from the pharyngeal orifice to the tympanic orifice in both positions. At the cartilaginous portion, the air space in the ET lumen was larger in the sitting position than in the recumbent position. At the bony portion of the ET, there did not seem to be any significant difference in the results of measurement in both cases. CONCLUSION: For the first time, we were able to obtain reconstructed images of patulous ET patients in the sitting position and to compare the findings with those of the recumbent position. We suggest that this method is useful for better understanding of the ET and ET related diseases such as patulous ET. PMID- 12753984 TI - Neurotrophin-3 modifies potassium currents in isolated inner hair cells from guinea-pig cochlea. AB - OBJECTIVE: Neurotrophins elicited short-term glutamate release from the presynaptic locus. The aim of this study was to investigate short-term effects of neurotrophin-3 (NT-3) on the membrane current of inner hair cells (IHCs). METHODS: IHCs were isolated from the guinea-pig cochlea. Membrane currents were measured by conventional whole-cell voltage-clamp recordings. NT-3 was dissolved in a standard external solution and applied to the IHCs under pressure using pipettes. RESULTS: Six out of eight IHCs demonstrated a suppression of K currents by extracellular application of NT-3. At +60 mV, the amplitudes of the outward current in the control and NT-3 solutions were 5.6+/-1.3 and 4.2+/-1.0 nA, respectively. NT-3 suppression was voltage-independent. One cell showed an immediate suppression with NT-3 and the following potentiation during washing with standard saline. CONCLUSION: NT-3 suppressed the amplitude of K current on IHCs, suggesting that neurotrophins are capable of potentiating the hair cells' excitability. Neurotrophins may have a therapeutic value in the prevention and treatment of hearing impairment caused by inner ear damage. PMID- 12753985 TI - Submucous electrocautery following submucous resection of turbinate bone--a rationale of surgical treatment for allergic rhinitis. AB - OBJECTIVE: Since the majority of the key elements such as trigeminal nerves, parasympathetic nerves, nasal glands, and blood vessels targeted by histamine and leukotrienes are found in the lamina propria of the nasal mucosa, its selective electrocautery has a rationale to improve the symptoms of allergic rhinitis with preservation of epithelial layer. To achieve the above goal, we performed the submucous electrocautery of the lamina propria (SECLP) following the submucous resection of the inferior turbinate bone (submucous turbinectomy: SMT). This paper discusses the efficacy of this procedure for the patients with persistent perennial allergic rhinitis. METHODS: An intranasal initial incision was made along the piriform aperture. The mucoperiosteum was elevated from the inferior turbinate bone followed by its complete resection. The SECLP was performed by applying a high-frequency coagulation current with a ball tip electrode, which was inserted into the mucoperiosteal sack after the completion of the SMT and was drawn forward on the medial surface of the mucoperiosteum with drawing a wavy line. We performed this surgery in 43 patients with perennial allergic rhinitis who were refractory to pharmacotherapy or were reluctant to take medicine. Symptoms, macroscopic intranasal findings, the results of allergic tests, nasal resistance, mucociliary function with saccharin, and the number of mast cells were compared pre- and postoperatively. RESULTS: The patients exhibited satisfactory improvement in symptoms only with a few crust formations. The macroscopic intranasal findings and allergic tests improved after surgery. Saccharin transport time remained normal. The number of anti-tryptase positive mast cells significantly decreased in the epithelial layer and in the superficial layer of the lamina propria of the postoperative inferior turbinate mucosa. CONCLUSION: The SECLP following the SMT is evaluated to be a useful surgical modality for allergic rhinitis with preservation of the nasal mucosal function. PMID- 12753986 TI - 'The columellotomy modification' of transseptal hypophysectomy. AB - OBJECTIVE: Although the transseptal approach to the sella turcica is the most common method by which the pituitary tumors are resected, it is not suitable for the noses with narrow nostrils, precluding the placement of the neurosurgical speculum. In this paper, as a solution for this problem, 'the columellotomy modification' of the transseptal approach is suggested and described together with the results in five patients. METHODS: In five patients, the distance between the outer edges of the two blades of the neurosurgical speculum was measured after positioning the neurosurgical speculum through the hemitransfixion incision, and after positioning the neurosurgical speculum through 'the columellotomy' incision. The increase in distance after hemitransfixion incision with respect to the width of the nostril was statistically compared with the increase in distance after 'columellotomy' incision with respect to the width of the nostril. The postoperative complaints and cosmetic results were evaluated. RESULTS: By paired-samples t-test, the comparison between the values of increase revealed a statistically significant difference (P=0.011). A complaint of numbness on the columellar skin was noted in three of the five patients. Cosmetic results were satisfactory. CONCLUSION: 'The columellotomy modification' offers a wide, direct and fast exposure to the sella turcica without a cosmetic disadvantage in noses with narrow nostrils. PMID- 12753987 TI - Beneficial effect of low-dose peritonsillar injection of lidocaine-adrenaline before tonsillectomy. A placebo-controlled clinical trial. AB - OBJECTIVE: Most studies investigating the effect of preincisional injection of local anaesthetic with adrenaline in tonsillectomy have used rather large doses and an inter-individual study design. They are inconclusive regarding the effect on post-tonsillectomy pain but have shown that the peroperative blood loss is reduced. However, side effects to high adrenaline doses are common. In the present study, the effect of injecting a small dose of lidocaine-adrenaline was investigated by using an intra-individual study design. METHODS: In this randomised double-blind study 52 patients (33 women and 19 men) received unilateral preincisional injection of 3-ml lidocaine-adrenaline before tonsillectomy. RESULTS: the difference in blood loss was highly significant with a more than 50% reduction in peroperative blood loss in the infiltrated side. Time to achieve haemostasis for one tonsil side was reduced from 6.0 to 4.0 min. No side effects were observed. At the operation day (mean 4.9 h after operation), the visual analogue scale for pain at the side infiltrated with local anaesthetics was 2.78 compared to 4.00 in the control side (P=0.05). CONCLUSION: Low-dose injection of lidocaine-adrenaline before tonsillectomy reduces blood loss and has a small but significant beneficial effect on early postoperative pain. Furthermore our results indicate that operation time is reduced. We therefore recommend subcapsular injection of 3 ml of 1% lidocaine with 1/200.000 adrenaline under each tonsil before tonsillectomy. PMID- 12753989 TI - The presence of large lymph node metastasis as a prognostic factor of papillary thyroid carcinoma. AB - OBJECTIVE: Papillary thyroid carcinomas often metastasize to regional lymph nodes, but the significance of lymph node metastasis as a prognostic factor has not been established. This study was conducted to determine the survival rate in patients with and without large lymph node metastasis greater than 3 cm in the maximum diameter, which are N2 or N3 for other head and neck cancers. METHODS: The survival rate was determined by using Kaplan-Meier method in 67 patients with previously untreated papillary thyroid carcinoma who underwent radical surgery at the Department of Otolaryngology, Toyooka Hospital, between 1993 and 2000. RESULTS: The prognosis was significantly poor (P=0.004) in patients with large lymph node metastasis greater than 3 cm in the maximum diameter. No significant difference was noted in the prevalence of large lymph node metastasis greater than 3 cm in the maximum diameter between different T groups. CONCLUSION: Large lymph node metastasis was considered to be a poor prognostic factor of papillary thyroid carcinoma independent of the T classification. PMID- 12753988 TI - Reduced expression of p27 is correlated with progression in precancerous lesions of the larynx. AB - OBJECTIVES: Factors related to malignant transformation of laryngeal precancerous lesions remain largely unknown, so we investigated the relationship between the expression of p27 and precancerous laryngeal lesion. STUDY DESIGN: In this study we investigated the expressions of p27 and p53 protein in 56 cases with laryngeal precancerous or cancer lesions (20 cases of hyperplasia, 19 of dysplasia, and 17 of squamous cell carcinoma), and went on to evaluate the relationship between immunoreactivity of each of them and the histological findings. We also evaluated the correlation between immunoreactivity and proliferative activity with the aid of Ki-67 nuclear antigen staining. METHODS: A retrospective study was performed in 56 cases (20 with epithelial hyperplasia, 19 with epithelial dysplasia and 17 with laryngeal cancer). Immunohistochemistry was performed to investigate expression of p27, p53 protein and Ki-67 nuclear antigen staining, using the avidian-biotin-peroxidase complex technique. RESULTS: p27 immunostaining was observed in 12 out of 20 cases of hyperplasia (60%), six out of 19 cases of dysplasia (31%), and 2/17 (12%) of carcinoma. We found significant association between p27 immunostaining and the histological findings. On the other hand, p53 immunostaining was observed in 6/20 (30%) of hyperplasia, 3/19 (16%) of dysplasia, and 7/17 (41%) of carcinoma. No significant association was found between p53 and the histological findings. CONCLUSIONS: Our results revealed that immunohistochemical assessment of p27 in bioptic samples of laryngeal precancerous lesions might be useful in selective patients who should undergo a more specific follow-up evaluation. PMID- 12753990 TI - Management of hypopharyngeal and cervical oesophageal perforations. AB - BACKGROUND: Perforations of the hypopharynx and the cervical oesophagus are infrequent severe situations, which may even be life-threatening for patients. METHODS: We review seven cases of intraluminal perforations of the hypopharynx or cervical oesophagus treated at our department between 1999 and 2001. RESULTS: In this series of patients, foreign bodies were the main cause of perforation. In four cases, the treatment was surgical by means of a cervicotomy and/or thoracotomy and drainage; in the other three cases, conservative treatment was applied. In some cases, the morbidity was considerable but there were no mortalities. CONCLUSIONS: The treatment of perforations of the hypopharynx and the cervical oesophagus must be individualized and multidisciplinary. The early diagnosis of these perforations is an important factor for prognosis. PMID- 12753991 TI - Subjective pulsatile tinnitus associated with extensive pneumatization of temporal bone. AB - Pulsatile tinnitus (PT) although an infrequent otologic symptom. PT can be objective (auscaltable) or subjective (non-auscultable). It has been suggested that subjective PT could occasionally be associated with vascular disorders such as arteriovenous malformation, traumatic or spontaneous carotico-cavernous fistula, intracranial aneurysms, vascular tumors of the temporal bone and cerebellopontine angle, fibromuscular dysplasia, cervical venous hums and high jugular bulb. To our literature knowledge, it has not been reported subjective PT due to extensive pneumatization of temporal bone around internal carotid artery (ICA). In this report, we present a case of subjective PT, which was caused by resonance due to extensive pneumatization of temporal bone particularly peripheral to the ICA. PMID- 12753992 TI - Otogenic pneumocephalus management with butyl 2-cyanoacrylate. AB - The existence of air within the skull has been called pneumocephalus. Treatment of pneumocephalus is mostly managed surgically, especially in a tension pneumocaephalus when the pressure has to be alleviated. Dural defect can be treated with the suture technique and tissue adhesives such as butyl 2 cyanoacrylate (B 2-CA). Otogenic pneumocephalus due to dural defect can be managed with B 2-CA tissue adhesive cheap, safely, and effectively without complication. We thought it would be of interest to present a case of pneumocephalus due to traffic accident 12 years ago and managed with B 2-CA. It is concluded that B 2-CA fixation can be alternative for dural defect closure with suturing or other tissue adhesives. PMID- 12753993 TI - Laryngeal solitary multiple mucosal neuromas without multiple endocrine neoplasia (MEN) type 2B. AB - The first case of laryngeal solitary multiple mucosal neuromas (MMN) was reported. A 73-year-old man who had complained of hoarseness and abnormal prickly sensation in the throat for 3 months visited our hospital. Many small whitish yellow nodules were observed in the laryngeal mucosa from the right arytenoid to the interarytenoid region. Using laryngomicroscopy biopsy was performed twice. The pathological study showed neurogenic nodular lesions consisting of regularly arranged nerve bundles with many axons and Schwann cell proliferation, and so MMN was finally diagnosed. As MMN is regarded as a constant component of multiple endocrine neoplasia (MEN) type 2B, screening of MEN type 2B was performed, however, they were all within normal limits. This case was, therefore, diagnosed as a laryngeal solitary MMN without MEN type 2B. PMID- 12753994 TI - A case of orbital emphysema associated with internal laryngocele. AB - Orbital emphysema (OE) is usually the result of a fracture of lamina papyrecea or maxillary roof allowing air to pass from the sinuses into the orbit. OE without evidence of any significant trauma is a rare occurrence. Thorough literature search revealed that no case of OE due to the rupture of internal laryngocele has been reported. A case is reported here of bilateral orbitopalpebral emphysema associated with internal laryngocele, in the absence of facial skeleton trauma, in a healthy young adult male. It should be kept in mind that laryngocele rupture would be one of the underlying causes of OE or OE would be the presenting symptom of laryngocele rupture. PMID- 12753995 TI - Epithelial-myoepithelial carcinoma of the parotid gland. AB - We report a rare case of epithelial-myoepithelial carcinoma (EMC) of the parotid gland. A 70-year-old man presented with a 4-months-history of right-sided subauricular swelling. Computed tomographic scans revealed a well-defined mass with cystic lesion, measuring about 40 mm in diameter, in the right parotid gland. Because the tumor occupied superficial lobe, he underwent superficial parotidectomy with preservation of the facial nerve. On the basis of the histological and immunohistochemical findings, the tumor was diagnosed as EMC. His post-operative course was uneventful, and he is currently free from disease 6 months after surgery. Diagnosis, clinical behavior and treatment of EMC are reviewed from perusal of the literature. PMID- 12753996 TI - De novo papillary squamous carcinoma of the lacrimal sac. AB - We report a rare case of a papillary squamous carcinoma of the lacrimal sac diagnosed at first presentation. The initial presentation of the patient discussed here mimicked that of a lacrimal sac mucocele. This report highlights the clinicopathological characteristics of this tumour, as well as the multidisciplinary approach required in its management. PMID- 12753997 TI - Unusual cases of cervical nerves schwannomas: phrenic and vagus nerve involvement. AB - Benign neurogenic tumors (neurilemmoma) arising from the cervical phrenic or vagus nerve are relatively rare. These lesions are benign, asymptomatic and incidentally found. We describe two cases considering different surgical techniques adopted. In the case of phrenic nerve schwannoma we performed a total excision of the tumor including the maternal nerve fiber to prevent tumor recurrence, also in regard to the already present hemidiaphragm palsy. On the other hand in second case, in which the vagus nerve was involved, we proposed a microsurgical approach by monitoring nerve function in order to minimize nerve damage. PMID- 12753999 TI - Real time breath and headspace analysis of flavour volatiles. AB - An overview is presented of the principle, scope and major applications to date of the use of atmospheric pressure chemical ionisation mass spectrometry (APCI MS) for monitoring the kinetics of release of flavour volatiles in real time, principally from breath during eating. The technique is rapid, quantitative, sensitive to the ppb level and can be used to monitor the vast majority of flavour volatiles. Advances made during the last 5 years in our understanding of factors affecting flavour release, particularly when conducted simultaneously with sensory evaluation, are contributing increasingly to more efficient product development in the food and flavour industry and to the design of flavour systems with desired dynamic flavour characteristics. Real time APCI-MS headspace data may be used to validate mathematical modelling of flavour release. It is proposed that these advances may be applied with similar benefits in the pharmaceutical industry, particularly in the improvement of the flavour acceptability of orally administered drugs. PMID- 12754000 TI - Liposomes increase skin penetration of entrapped and non-entrapped hydrophilic substances into human skin: a skin penetration and confocal laser scanning microscopy study. AB - Liposomes have been extensively studied and suggested as a vehicle for topical drug delivery systems. However, the mechanism by which liposomes deliver drugs into intact skin is not fully understood. In the present study, we have tried to understand the mechanism of transport of hydrophilic drugs into the skin using liposomes. The effect of separation of the non-entrapped, hydrophilic fluorescent compound, carboxyfluorescein (CF), from liposomally entrapped CF was investigated by measuring the penetration of CF across human skin under non-occlusive conditions in vitro using Franz diffusion cells. The fluorescent dye, CF, was incorporated into the liposomes and applied onto the skin. After a 6 and 12h incubation period, the amount of CF in the epidermal membrane and the full thickness skin was determined by fluorescence spectroscopy or by confocal laser scanning microscopy (CLSM). The liposomal formulation containing CF both inside and outside the vesicles showed statistically enhanced penetration of CF into the human stratum corneum (SC) as compared to the formulations containing CF only outside of the liposomes and CF in Tris buffer. The CLSM results revealed that the formulation in which CF was present outside the liposomes showed bright fluorescence intensity in the SC and very weak fluorescence in the viable epidermis. However, the CF in Tris buffer failed to show any fluorescence in the viable epidermis. The results indicated that phospholipid vesicles not only carry the entrapped hydrophilic substance, but also the non-entrapped hydrophilic substance into the SC and possibly into the deeper layers of the skin. PMID- 12754001 TI - Scanning electron microscopy and atomic force microscopy imaging of solid lipid nanoparticles derived from amphiphilic cyclodextrins. AB - Scanning electron microscopy (SEM) and atomic force microscopy (AFM) have been applied to the imagery of solid lipid nanoparticles (SLNs) formulated from an amphiphilic cyclodextrin, 2,3-di-o-alkanoyl-beta-cyclodextrin, beta-CD21C6. Comparison of the results shows that the vacuum drying technique used in sample preparation for SEM causes shrinkage in the size of the SLNs, whereas the deposition method used for AFM causes the SLNs to form small clusters. The hydrodynamic diameter determined from photon correlation spectroscopy (PCS) is 359+/-15 nm and the zeta potential is -25 mV. PMID- 12754002 TI - Mechanistic appraisal of the effects of some protease inhibitors on ciliary beat frequency in a sequential cell culture system of human nasal epithelium. AB - The aim of this study was to investigate the suitability of a sequential monolayer-suspension culture system as a model to screen subacute effects of drug excipients on ciliary beat frequency (CBF). The CBF of the cultured cells was measured by computerized microscope photometry. Protease inhibitors (puromycin, bestatin, bacitracin, actinonin and thiomersal) were used as model compounds and the mechanisms of ciliary inhibition were investigated by probing the involvement of arachidonic acid metabolism, guanylate cyclase (cGMP), protein kinase C (PKC) and adenosinetriphosphate (ATP) inhibition. Bestatin concentration-dependently reduced CBF by inhibiting arachidonic acid metabolism, cGMP, PKC and endogenous ATP consumption. Thiomersal and DMSO used for dissolving actinonin reduced CBF (P<0.05) via a non-specific mechanism. Bacitracin (8 mM) and puromycin (135 mM) had no effect on CBF after acute exposure (15-30 min) (P>0.05), but significantly reduced the CBF by approximately 15.0% following daily 15-min exposure for 1 week. This study shows that (i) sequential monolayer-suspension culture system is a valid model to screen both acute and subacute effects of drug excipients on CBF; and (ii) bacitracin, puromycin and actinonin are more cilio-compatible than bestatin and thiomersal and as such are more potentially useful nasal absorption enhancer from ciliotoxicity perspective. PMID- 12754003 TI - Powdered cellulose as excipient for extrusion-spheronization pellets of a cohesive hydrophobic drug. AB - This study compared a powdered cellulose (PC) and a microcrystalline cellulose (MCC) as sole excipients in the preparation of furosemide pellets by extrusion spheronization. Pellets prepared with PC and 25 or 50% furosemide showed smaller mean size, a broader particle size distribution, similar sphericity, greater surface roughness and higher friability than equivalent pellets prepared with MCC. Furosemide release rate was markedly higher from PC pellets, which may be attributable to their higher micropore volume. PMID- 12754004 TI - Physicochemical properties to determine the buoyancy of hollow microspheres (microballoons) prepared by the emulsion solvent diffusion method. AB - Hollow microspheres (microballoons) floatable on JPXIII No.1 solution were developed as a dosage form capable of floating in the stomach. Hollow microspheres were prepared by the emulsion solvent diffusion method using enteric acrylic polymers with drug in a mixture of dichloromethane and ethanol. It was found that preparation temperature determined the formation of cavity inside the microsphere and the surface smoothness, determining the floatability and the drug release rate of the microballoon. The correlation between the buoyancy of microballoons and their physical properties, e.g. apparent density and roundness of microballoons were elucidated. The drug loading efficiency of microballoons with various types of drug was investigated and correlated to the distribution coefficient of drug between dichloromethane and water. The optimum loading amount of riboflavin in the microballoon was found to impart ideal floatable properties to the microballoons. On the other hand, little entrapment was observed for aspirin due to the low distribution coefficient; however, entrapment improved to some extent upon reduction of the pH of the process. PMID- 12754005 TI - Formulation of intravenous carbamazepine emulsions by SolEmuls technology. AB - Oil in water (O/W) emulsions for parenteral nutrition can be employed as intravenous (i.v.) carriers for drugs that are poorly soluble in water and in oil by localising the drug in the interfacial lecithin layer, e.g. Amphotericin B emulsions. By now, the emulsion production required organic solvents. SolEmuls technology localises the drug in the interfacial layer by a solvent-free high pressure homogenisation process. SolEmuls was applied to produce Carbamazepine emulsions at increasing drug concentrations from 0.5 to 10mg/ml. Drug powder and Lipofundin emulsion were mixed and homogenised at 1500bar. Characterisation of emulsions and short-term stability were performed by photon correlation spectroscopy (PCS) and laser diffractometry. Drug incorporation (absence of non dissolved drug crystals) was investigated by light microscopy and a centrifugation test. The emulsions were physically stable and complete drug dissolution is possible up to 3mg/ml. Up to 10mg/ml drug hybrid dispersions of emulsion droplets and ultrafine nanocrystals were obtained. Both, emulsions and hybrid dispersions are suitable as i.v. injectables regarding size and stability. PMID- 12754006 TI - Enteric coating of soft gelatin capsules by spouted bed: effect of operating conditions on coating efficiency and on product quality. AB - The present study was conducted in order to analyze the viability of the spouted bed process for application of a gastric-resistant coating to soft gelatin capsules. The variables investigated were: included angle of conical base, (gamma), the relation between the feed mass flow rate of the coating suspension and the feed mass flow rate of spouting gas (W(s)/W(g)); the ratio between the flow rate of the spouting gas and the flow rate at minimum spouting condition (Q/Q(ms)); the mass of capsules in the bed (M(0)), and the capsule's size. The product quality was measured by disintegration tests, traction x deformation tests, image analysis and by the evaluation of the coating mass distribution and shape factor variation during the coating operation. The experiments were performed in a spouted bed with a column diameter of 200 mm and included a conical base angle of 40 degrees. The best coating efficiency values were obtained for M(0)=300 g. Coating efficiency tended to increase with increasing W(s)/W(g) ratio. Disintegration tests showed that the gastric-resistant effect was obtained with a coating mass of 3.86 mg/cm(2). The shape factor increase during the coating operation. The capsule's coating mass distribution tended to maintain the original distribution. PMID- 12754007 TI - Rheological study on mucoadhesivity of some nasal powder formulations. AB - Various powder mixtures were used to administer insulin via the nasal route: a co spray dried mixture of Amioca starch and Carbopol 974 P (1/3), drum dried waxy maize starch and Carbopol 974 P (9/1), maltodextrin DE38/Carbopol 974 P (9/1) and pure drum dried waxy maize starch. Oscillatory rheology is performed to study and compare the viscosity, elasticity and mucoadhesivity of these powder formulations. There was no rheological synergism detectable with the co-spray dried mixture of Amioca starch and Carbopol 974 P (1/3), drum dried waxy maize starch and Carbopol 974 P (9/1) and maltodextrin DE38/Carbopol 974 P (9/1). Interaction due to entanglements was seen with drum dried waxy maize starch (100%). The differences in nasal bioavailability between the different carriers could be explained by differences in G' (storage modulus, elasticity) and G" (loss modulus, viscosity) values. The formulation giving the highest bioavailability, provided also the highest G' and G" values. PMID- 12754008 TI - Influence of menthol and pressure-sensitive adhesives on the in vivo performance of membrane-moderated transdermal therapeutic system of nicardipine hydrochloride in human volunteers. AB - A membrane-moderated transdermal therapeutic system of nicardipine hydrochloride was developed using 2% w/w hydroxypropylcellulose (HPC) gel as a reservoir system containing 5% w/w of menthol as a penetration enhancer. The permeability flux of nicardipine hydrochloride through the ethylene vinyl acetate (EVA) copolymer membrane was found to increase with an increase in vinyl acetate content in the copolymer. The effect of various pressure-sensitive adhesives (MA-31, MA-38 or TACKWHITE A 4MED on the permeability of nicardipine hydrochloride through EVA 2825 membrane (28% w/w vinyl acetate) or EVA 2825 membrane/skin composite was also studied. The results showed that nicardipine hydrochloride permeability through EVA 2825 membrane coated with TACKWHITE A 4MED/skin composite was higher than that coated with MA-31 or MA-38. Thus, a new transdermal therapeutic system for nicardipine hydrochloride was formulated using EVA 2825 membrane coated with a pressure-sensitive adhesive TACKWHITE A 4MED, and 2% w/w HPC gel as reservoir containing 5% w/w of menthol as a penetration enhancer. In vivo studies in healthy human volunteers indicated that the TTS of nicardipine hydrochloride, designed in the present study, provided steady-state plasma concentration of the drug with minimal fluctuations for 26h with improved bioavailability in comparison with the immediate release capsule dosage form. PMID- 12754009 TI - Assessment of a controlled release hydrophilic matrix formulation for metoclopramide HCl. AB - Metoclopramide HCl showed controlled release behavior when embedded in a hydrophilic matrix of chitosan and sodium alginate. The in vitro release data was found to be first order according to the Higuchi mechanism. An in vivo evaluation of the metoclopramide controlled release matrix on six male volunteers was carried out. The plasma samples were analyzed using a high-performance liquid chromatography (HPLC) method using a mobile phase of acetonitrile:acetic acid (30:70), with the pH adjusted to 4.7, a reverse phase Hypersil BDS Phenyl column (4 microm, 250 x 4 mm) and the detection was performed at 305 nm. The controlled release formula was found to be effective in delaying absorption (t(max) 4.5h as compared to 1.2h), reducing the peak plasma concentrations (C(max) 63.4 ng/ml as compared to 95.9 ng/ml) and maintaining higher concentrations during the elimination phase when compared to the immediate release formula. This proves the suitability of the suggested system for further studies. PMID- 12754010 TI - Quality evaluation of generic drugs by dissolution test: changing the USP dissolution medium to distinguish between active and non-active mebendazole polymorphs. AB - Mebendazole is practically insoluble in water and studies of its polymorphism has led to the identification and characterization of three polymorphic forms (A, B, C) displaying solubility and therapeutic differences that show that polymorph C is therapeutically favored. The objective of this study was to adjust the USP dissolution test for mebendazole so that it was able to distinguish between the dissolution properties of three mebendazole polymorphs. This would provide generic manufacturers with one more test to ensure that the therapeutically active polymorph C is used. The results obtained in this study show that the USP dissolution test conditions were not able to distinguish between the dissolution properties of completely dispersed mebendazole polymorphs with comparable particle sizes. When sodium lauryl sulfate was removed from the dissolution medium, the percentage dissolved versus time profiles changed so that polymorph C dissolved faster (70% within 120 min) compared to polymorph B (37% within 120 min) and polymorph A (20% within 120 min). PMID- 12754011 TI - The effect of polymer blends on release profiles of diclofenac sodium from matrices. AB - The purpose of this study was to evaluate the effect of polymer blends on the in vitro release profile of diclofenac sodium. Several controlled release matrices of diclofenac sodium with different proportions of hydroxypropyl methylcellulose (HPMC; viscosity grade 60 and 500 mPa.s), carbopol 940 and lactose as a water soluble filler were prepared. The results showed that when HPMC (viscosity grade 60 mPa.s) alone was used as matrix former, diclofenac sodium was released fast but the release rate became slower with HPMC (viscosity grade 500 mPa.s) at higher polymer/drug ratios (more than 0.8:1). However in lower polymer/drug ratios (lower than 0.7:1) the release rate still was fast. The results showed that carbopol can extend the release time appreciably but the release profiles had considerable fluctuations, and drug release in first hours was slow but increased appreciably with time at the end of profiles. When an appropriate blend of HPMC (viscosity grade 60 or 500 mPa.s) and carbopol 940 was used, the drug release became more uniform and its kinetic approached to zero order and release fluctuations were diminished. The results with these polymer blends showed that it is possible to reduce the total amounts of polymer in each formulation. According to kinetic analysis data, drug release from these matrix tablets did not follow Fick's law of diffusion and the results were in agreement with the earlier reports. PMID- 12754012 TI - Aerosol flow reactor method for synthesis of drug nanoparticles. AB - An aerosol flow reactor method, a one-step continuous process to produce nanometer-sized drug particles with unimodal size distribution, was developed. This method involves first dissolving the drug material in question into a suitable solvent, which is then followed by atomising the solution as fine droplets into carrier gas. A heated laminar flow reactor tube is used to evaporate the solvent, and solid drug nanoparticles are formed. In this study, the effect of drying temperature on the particle size and morphology was examined. A glucocorticosteroid used for asthma therapy, beclomethasone dipropionate, was selected as an experimental model drug. The geometric number mean particle diameter increases significantly with increasing reactor temperatures due to formation of hollow nanoparticles. Above 160 degrees C, however, further increase in temperature results in decreasing particle size. The produced nanoparticles are spherical and show smooth surfaces at all studied experimental conditions. PMID- 12754014 TI - Is total arterial myocardial revascularization with composite grafts a safe and useful procedure in the elderly? AB - OBJECTIVE: The aim of the study was to evaluate the mid-term results of total arterial myocardial revascularization (TAMR) with composite grafts in patients older than 70 years when compared to standard CABG technique, since the usefulness of TAMR in the elderly has not been demonstrated yet. METHODS: A prospective randomized study was designed with the following end-points: post operative complications, death, recurrence of angina, graft occlusion, any cardiac event and reinterventions. One hundred and eighty-eight patients older than 70 years were enrolled and assigned to Group 1(G1)=94 pts, for total arterial revascularization or Group 2(G2)=94 pts, for standard CABG (LITA on LAD plus additional saphenous veins). The groups were comparable in terms of pre operative characteristics and Euroscore (mean: G1=8.4 vs. G2=8.2). RESULTS: No differences between the groups were observed in terms of mean number of grafted vessels (G1=2.1 vs. G2=2.3), mean aortic cross-clamping time (G1=34+/-8 vs. G2=33+/-6min), mechanical ventilation time (G1=23+/-4 vs. G2=22+/-4hr), ICU stay (G1=40+/-10 vs. G2=39+/-9hr), post-operative complications and hospital mortality (G1=5.3% vs. G2=4.2%). At a mean follow-up of 12+/-4 months, cumulative incidence of angina recurrence was 2.1% in G1 vs. 11% in G2 (P=0.021). Angiographic evaluation showed 98.2% arterial patency in G1 vs. 86% saphenous vein graft patency in G2 (P<0.001). Multivariate analysis identified conventional CABG surgery as independent predictor of angina recurrence, graft occlusion and late cardiac events. CONCLUSIONS: Total arterial revascularization with composite grafts proved to be a safe and effective procedure also in the elderly. Composite arterial grafts provided superior clinical outcome with a lower rate of angina recurrence, graft occlusion and late cardiac events when compared to conventional CABG strategy. PMID- 12754015 TI - Flow simulation of the intracoronary shunt tube for off-pump coronary artery bypass. AB - INTRODUCTION: To simulate blood flow provided by the intracoronary shunt tube, and to clarify whether this method is actually suitable for Off-pump coronary artery bypass (OPCAB), we investigated the efficacy of the intracoronary shunt tube in a theoretical model on the basis of fluid dynamics. METHODS: Fluid dynamics analysis was performed to simulate flow decrease after attachment on an intracoronary shunt model. RESULTS: The flow ratio in the case of turbulent flow is in proportion to the ratio of the inner diameter to the third power, and that in the case of laminar flow is in proportion to the ratio of the inner diameter to the sixth power. When this analysis is applied to commercial shunt tubes, coronary flow was estimated as approximately 2-14% of pre-attachment flow in turbulent flow, and only less than 0.1% in laminar flow. CONCLUSIONS: This result suggests that use of intracoronary shunt tubes in OPCAB may rarely contribute to maintenance of coronary flow, and they should be used carefully, especially in a jeopardized coronary artery. PMID- 12754016 TI - Superior myocardial protection with nicorandil cardioplegia. AB - OBJECTIVE: The ATP-sensitive potassium channel (K(ATP)) activator nicorandil used as cardioplegic agent may protect the left ventricle during cardiac arrest. Nicorandil in cold blood was compared with standard hyperkalemic blood and crystalloid cardioplegia. METHODS: Twenty-one pigs were randomly assigned to three groups: (1) cold hyperkalemic crystalloid (n=7); (2) cold hyperkalemic blood (n=7); and (3) nicorandil as cardioplegia in cold blood (n=7). Left ventricular mechanical performance, pressure-volume area (PVA) and myocardial oxygen consumption (MVO(2)) were measured before and at 1 and at 2 h after 60 min of cold global ischemia on cardiopulmonary bypass using intraventricular pressure volume conductance catheters, coronary flow probes and O(2)-content difference. RESULTS: The slope (M(w)) of the stroke work end-diastolic volume relationship, the preload recriutable stroke work relationship, was unchanged after ischemia in the nicorandil group, but was reduced to averaged 62.5% (standard deviation 14) of baseline values in both hyperkalemic perfusions (P<0.05). The slope of the MVO(2)-PVA relationship was unchanged after nicorandil cardioplegia while the slope after hyperkalemic blood and crystalloid cardioplegia increased with 33% (P<0.02) and 52% (P<0.02) of baseline values, respectively. CONCLUSIONS: Nicorandil as sole cardioplegic agent in cold blood given intermittently preserves left ventricular contractility and myocardial energetics significantly better than traditional forms of cardioplegia after cardiac arrest. PMID- 12754017 TI - Fast track recovery of high risk coronary bypass surgery patients. AB - OBJECTIVE: Fast track recovery protocols on younger, low risk patients result in shorter hospital stays and decreased costs. However, data is lacking on the impact of these protocols on high risk patients based on an objective scoring system. METHODS: In this study, a high risk cohort of patients (EuroSCORE >or=6, n=158) was compared with a low risk cohort of patients (EuroSCORE <6, n=1004) to define the safety and efficacy of fast track recovery among high risk patients. A standard perioperative data is collected prospectively for every patient. RESULTS: Time to extubation was longer in the high risk group (299+/-253 vs. 232+/-256min; P=0.003), but intensive care unit (ICU) stay (25.6+/-28.7 vs. 21.5+/-9.4h; P=ns), and postoperative length of stay (5.8+/-2.4 vs. 5.6+/-2.7 days; P=ns) was similar when compared with the low risk group. Of the high risk patients 81% were extubated within 6h, 87% were discharged from the intensive care unit within 24h, and 67% were discharged from the hospital within 5 days. Multiple regression analysis showed that any red blood cell transfusion (P=0.02), and cross clamp time >60min (P=0.03) were the predictors of delayed extubation (>or=6h) in the high risk group. The predictors of extended ICU stay were any red blood cell transfusion (P=0.0001), and peripheral vascular disease (P=0.05). Any red blood cell transfusion was the only predictor for mortality (P=0.02) and readmission to the hospital within the first 30 days (P=0.02) in this cohort of patients. CONCLUSIONS: This study confirms the safety and efficacy of fast track recovery protocol among high risk patients undergoing coronary artery bypass surgery. All patients are basically suitable for fast track recovery and the preoperative risk factors are poor predictors of prolonged ventilation, increased ICU and hospital stay. Red blood cell transfusion is associated with delayed extubation and discharge from the ICU, and increased mortality and hospital readmission rate. PMID- 12754018 TI - Logistic or additive EuroSCORE for high-risk patients? AB - OBJECTIVES: To assess whether the use of the full logistic European System for Cardiac Operative Risk Evaluation (EuroSCORE) is superior to the standard additive EuroSCORE in predicting mortality in high-risk cardiac surgical patients. METHODS: Both the simple additive EuroSCORE and the full logistic EuroSCORE were applied to 14,799 cardiac surgical patients from across Europe, of whom there were 4293 high-risk patients (additive EuroSCORE of 6 or more). The systems were compared for absolute prediction and discrimination (area under the receiver operating characteristic (ROC) curve). RESULTS: Actual mortality was 4.72%. The logistic model was closer to this than the additive model (4.84% (4.72 4.94) versus 4.21 (4.21-4.26)). Most of this difference was due to high-risk patients where actual mortality was 11.18% and predicted was 7.83% (additive) and 11.23% (logistic). Discrimination was similar in both systems as measured by the area under the ROC curve (additive 0.783, logistic 0.785). CONCLUSIONS: The additive EuroSCORE model remains a simple "gold standard" for risk assessment in European cardiac surgery, usable at the bedside without complex calculations or information technology. The logistic model is a better risk predictor especially in high-risk patients and may be of interest to institutions engaged in the study and development of risk stratification. PMID- 12754019 TI - Choice of a mechanical valve or a bioprosthesis for AVR: does CABG matter? AB - OBJECTIVE: Mechanical valves and bioprostheses are the commonly used devices in aortic valve replacement (AVR). Many patients with valvular disease also require concomitant coronary artery bypass grafting (CABG). We used a microsimulation model to provide insight into the outcomes of patients after AVR with mechanical valves and stented bioprostheses, with and without CABG, and to determine the age thresholds or age crossover points in outcomes between the two valve types. METHODS: We conducted a meta-analysis of published results after primary AVR with mechanical prostheses (nine reports, 4274 patients, 25,726 patient-years) and stented porcine bioprostheses (13 reports, 9007 patients, 54,151 patient-years) to estimate risks of valve-related events. A hazard ratio of 1.3 was used to incorporate the effect of CABG on long-term survival. Estimates were entered into a microsimulation model, which was then used to predict the outcomes of patients after AVR, with and without CABG. The model calculations were validated using a large data set from Portland, USA. RESULTS: For a 65-year-old male without CABG, the life expectancy (LE) was 11.2 and 11.6 years and the event-free life expectancy (EFLE) was 8.2 and 8.9 years, respectively, after implantation with mechanical valves and bioprostheses. The lifetime risk of at least one valve related event was 51 and 47%, respectively. The age crossover point between the two valve types, considering the above outcome parameters, was 59, 60 and 63 years, respectively. CABG reduced LE and consequently EFLE and lifetime risk of an event, but only minimally influenced the patient age crossover points. The model calculations showed good agreement with the Portland data. CONCLUSIONS: The currently recommended patient age for using a bioprosthesis (65 years) could be lowered further, irrespective of concomitant CABG. The trade-off between the reduced risks of bioprosthetic failure and of hemorrhage in mechanical valves, resulting from a lower LE, minimized the effect of CABG on the age crossover points between the two valve types. PMID- 12754020 TI - Left ventricular mass index in aortic valve surgery: a new index for early valve replacement? AB - OBJECTIVE: Increased left ventricular mass index has been associated with higher mortality. We analyze the effect of increased left ventricular mass index on outcomes in patients undergoing aortic valve replacement. METHODS: Echocardiographic left ventricular dimensions were used to calculate left ventricular mass index in 614 patients who underwent aortic valve replacement between June 1993 and November 2001. Left ventricular mass index was considered increased if higher than the value of the superior decile (277 g/m(2) in males and 251 in females). RESULTS: Mean left ventricular mass index was: 178+/-111 g/m(2), and increased index was considered in 9.9% of patients. Postoperative complications (low cardiac output syndrome, respiratory failure, arrhythmias, pneumonia and mediastinitis), median length of hospital stay: 12 days (6-57) versus 11 days (5-51), and in-hospital mortality (11.4, 3.2%, P<0.01) were higher in patients with increased left ventricular mass index. Multivariable analysis identified increased left ventricular mass index (odds ratio: 5.6; 95% confidence interval: 1.2-25.0; P=0.02) and other three variables: age (P=0.04), history of chronic renal failure (P=0.03) and cardiopulmonary bypass time (P=0.004), as independent predictors of early mortality. CONCLUSIONS: Increased left ventricular mass index is associated with an in-hospital adverse outcome and a significantly higher in-hospital mortality in patients undergoing aortic valve replacement. Outcomes in asymptomatic patients could be improved before a clinically significant increase in left ventricular mass index. Further studies should be performed to determine the usefulness of this index in selecting patients for earlier aortic valve replacement. PMID- 12754022 TI - Assessing the benefit of biological valve prostheses: cumulative incidence (actual) vs. Kaplan-Meier (actuarial) analysis. AB - OBJECTIVE: The standard method of analysing structural valve degeneration (SVD) of biological prostheses is the Kaplan-Meier method. In order to assess SVD with regard to competing risks (e.g. death particularly in elderly patients) cumulative incidence (actual analysis) was compared to Kaplan-Meier (actuarial analysis). METHODS: We retrospectively analysed 257 patients older than 60 years, who underwent mitral valve replacement with different biological prostheses between 1974 and 2000. Reoperation-free survival was determined, both according to Kaplan-Meier and cumulative incidence analysis. RESULTS: For the total group of patients older than 60 years, the 10- and 15-year freedom from reoperation was 79+/-5 and 55+/-8%, respectively, according to Kaplan-Meier and 90+/-2 and 83+/ 3% according to cumulative incidence analysis. For patients older than 65 years of age (n=170), Kaplan-Meier analysis revealed 85+/-7% freedom from reoperation at 10 years vs. 94+/-3% according to cumulative incidence analysis. For those between 60 and 65 years of age (n=87), Kaplan-Meier freedom from reoperation was 76+/-7% at 10 years and 48+/-9% at 15 years vs. 86+/-4 and 75+/-5% according to cumulative incidence analysis. CONCLUSIONS: Kaplan-Meier analysis overestimates the 10- and 15-year risk of SVD compared to cumulative incidence analysis, thus underestimating the benefit of biological valve replacement. Cumulative incidence analysis may lead to a more complete evaluation of risk and benefit and thus better patient management. PMID- 12754021 TI - Aortic valve replacement for aortic regurgitation and stenosis, in patients with severe left ventricular dysfunction. AB - OBJECTIVE: Aortic valve replacement for aortic valve stenosis (AS) and regurgitation (AR) in patients with severe left ventricular (LV) dysfunction contains an increased risk. Few data are available on the outcome of such patients. METHODS: Fifty-five consecutive patients with severe LV dysfunction (ejection fraction, EF; <30%) and aortic valve replacement for AS (n=35) or AR (n=20) were investigated between 1994 and 2001. EF was 25+/-5%, mean transvalvular gradient 26+/-6mmHg (AS), aortic valve area 0.66+/-0.18cm(2) (AS), cardiac index (CI) 2.4+/-0.9l/min/m(2), enddiastolic LV diameter (LVEDD) 64+/-8mm and endsystolic LV diameters (LVESD) was 55+/-3mm. Ninety percent of patients were in New York Heart Association (NYHA) functional class III/IV at admission to the hospital. Concomitant coronary artery bypass grafts (CABG) were performed in 14 patients. Follow-up examinations including chest X-ray, echocardiography, exercise testing, were performed among survivors. RESULTS: The survival rates for AS were: 1-year 76%, 2-year 68.8%, 5-year 64.2%; for AR: 1-year 94.4%, 2-year 86.5%, 5-year 74.2%. NYHA functional class improved from 90% in class III/IV to 45 (AR group) and 24% (AS group) at follow-up (P<0.02). The LVEDD decreased to 54+/-8mm after 1 year. The EF improved to 38+/-4 (AR group) and 40+/-5% (AS group) at follow-up. CONCLUSIONS: Despite severe LV dysfunction, increased 1-year mortality especially in the AS group, aortic valve replacement was associated with improved functional status, symptoms and EF in both groups and in most patients. We, therefore, conclude that aortic valve replacement in patients with severe LV dysfunction can be performed with acceptable risk. PMID- 12754023 TI - Premature failure of small-sized Shelhigh No-React porcine pulmonic valve conduit model NR-4000. AB - OBJECTIVE: Given the limited availability of small-sized cryopreserved pulmonary homografts, we implanted a series of Shelhigh No-React porcine pulmonic valve conduits (SPVC). The aim of this study was to evaluate the short-term performance following implantation. METHODS: From February 2000 to September 2000, the SPVC was implanted 25 times in 24 patients in the right ventricular outflow tract (RVOT) to correct congenital anomalies. The anatomical malformations were TOF/PA in eight patients, TGA/VSD/PS in four, truncus arteriosus in four, IAA/VSD/AS or AA in four, l-TGA/VSD in two and other in two. Age at operation was 2.8+/-3.9 years (mean+/-SD), including 12 patients under 1 year. The median conduit size was 14mm (range, 10-18). RESULTS: At a mean follow-up of 23+/-5 months, two late deaths (8%) have occurred. Although they were not primarily conduit related, both showed severe conduit stenosis. Twenty-one conduits (84%) showed mild to severe conduit stenosis, regurgitation or both. Two patients underwent balloon dilatation for distal conduit stenosis. Twelve conduits (48%) in 11 patients were removed at a median of 12 months (range, 2-18 months) due to RVOT obstruction in 11 and free conduit insufficiency with pseudoaneurysm in one. The typical findings of the explanted conduits were prominent intimal peel formation at the distal anastomosis without calcification. The actuarial freedom from reintervention at 18 months was 48+/-10%. CONCLUSIONS: Our experience of the SPVC with the diameter of 14mm or less has revealed a high incidence of distal conduit stenosis due to intimal peel formation resulting in early conduit failure. These findings have led us to abandon its use when other options are available. PMID- 12754024 TI - Increasing experience with integrated approach to pulmonary atresia with ventricular septal defect and major aortopulmonary collateral arteries. AB - OBJECTIVES: To validate the preliminary surgical results of 'integrated approach' to pulmonary atresia, ventricular septal defect (VSD), and multiple aortopulmonary collateral arteries by retrospective analysis of our center experience. METHODS: Between 01/94 and 03/02, 37 patients aged 22 days to 13 years underwent surgery for pulmonary atresia, VSD, and multiple aortopulmonary collaterals. Case selection was based on preoperative calculation of total neopulmonary arterial index (TNPAI), pulmonary arterial index (PAI), and pulmonary arteries-to-collateral arteries lung segment perfusion ratio (S(pa):S(ca)). The decision for a possible VSD closure during one-stage procedures was based on an intraoperative pulmonary flow study. Twenty-five patients with a TNPAI equal to or greater than 150 mm(2)/m(2) underwent primary unifocalization, irrespective of PAI and S(pa):S(ca). Conversely, 12 patients with a TNPAI less than 150 mm(2)/m(2) and hypoplastic (PAI less than 100 mm(2)/m(2)) dominant (S(pa):S(ca) greater than 1) pulmonary arteries received a first-stage right ventricular outflow tract reconstruction, followed by unifocalization and repair (i.e. VSD closure) in nine cases. RESULTS: Among 34 patients who received total unifocalization, the overall repairability rate was 85% (first instance repairs: n=27; delayed VSD closure: n=2; 95% confidence interval, CI: 73-97%), with a survival rate at 7 years of 81%. Repaired survivors (n=26) are asymptomatic (n=22) or mildly symptomatic (n=4) at a follow-up interval of 43+/-28 months, with a 0.48+/-0.2 mean haemodynamic right ventricular/left ventricular pressure ratio, whereas palliated ones are waiting for either repair (n=3) or catheter study (n=2). Analysis of results has shown the following: (1) 100% (34/34 cases) feasibility of one-stage unifocalization in patients with a preoperative TNPAI equal to or greater than 150 mm(2)/m(2), whereas combined repairability rate was 79% only (95% CI: 65-93%); (2) 100% (12/12 cases) fulfillment of criteria for second-stage repairability (acquired TNPAI greater than 150 mm(2)/m(2)) in all patients treated with right ventricular outflow tract reconstruction; and (3) 93% (95% CI: 83-100%) overall accuracy of intraoperative flow study in predicting either postrepair mean pulmonary arterial pressure (VSD closed: n=23) or balanced pulmonary to systemic blood flow ratio (VSD left open: n=4). CONCLUSIONS: Increasing experience with 'integrated approach' to pulmonary atresia, VSD, and multiple aortopulmonaty collaterals has confirmed the preliminary results of our surgical series. The pulmonary flow study remains the most accurate intraoperative test for successful management of VSD during unifocalization procedures PMID- 12754025 TI - Right ventricle-to-pulmonary artery shunt versus modified Blalock-Taussig shunt in the Norwood procedure for hypoplastic left heart syndrome - influence on early and late haemodynamic status. AB - OBJECTIVE: The aim of this study was to assess changes in early and late haemodynamic status after the Norwood procedure (NP), caused by the implementation of right ventricle-to-pulmonary artery shunt (RV-PA). METHODS: A consecutive series of 68 children with hypoplastic left heart syndrome underwent NP: Group 1 (n=31) with the application of a modified Blalock-Taussig shunt and Group 2 (n=37) with RV-PA. Haemodynamic data from the early postoperative period (72 h after the operation) and cardiac catheterisation data, as well as blood tests before the hemi-Fontan procedure (HF) were analysed. Univariate (chi(2) test, Mann-Whitney's and Student's t-tests) and multiple regression analysis were carried out. RESULTS: In Group 1, circulatory collapse requiring resuscitation occurred in 15 (48.4%) children, within 72 h after the procedure. The resuscitation was unsuccessful in nine (29%) cases. The operative mortality (30 days) was 35%. In Group 2, two (5%) children died within the early and two (5%) within the late postoperative period. The postoperative course in the remaining children from Group 2 was uneventful. In Group 2 there was a significantly higher mean diastolic pressure after NP (P<0.05). The arterial pulse pressure after NP was significantly lower in Group 2 (P<0.05). Before HF, the application of RV-PA was associated with a lower Qp:Qs ratio (P=0.020), lower aortic pulse pressure (P=0.004) and lower aortic oxygen saturation (P=0.039). CONCLUSIONS: A stable haemodynamic status due to independent coronary perfusion, higher diastolic and lower pulse pressure is the most advantageous effect of RV-PA, resulting in a lower mortality and morbidity after NP. A lower Qp:Qs ratio eliminates the danger of the ventricular volume overload and ensures good conditions for the development of the pulmonary circulation before HF. PMID- 12754026 TI - Mechanical limitation of pulmonary blood flow facilitates heart transplantation in older infants with hypoplastic left heart syndrome. AB - OBJECTIVES: Progression of pulmonary vascular disease limits heart transplantation for hypoplastic left heart syndrome (HLHS) to early infancy. Our objective was to assess the impact of bilateral pulmonary artery banding (PAB) on the operative courses of HLHS infants transplanted at ages older than 4 months. METHODS: Courses of all HLHS patients in our center who remained listed to age >or=120 days before heart transplantation were assessed. Patients undergoing transplantation after standard management (control group) were compared to patients having a prior pulmonary blood flow limiting procedure (PAB group). RESULTS: Of 16 identified patients, one crossed over to stage I Norwood on day 185 and died post-operatively. Fifteen patients were transplanted at age >or=120 days (control group n=9, PAB group n=6). Four PAB patients had open PA band placement. Two PAB patients underwent experimental percutaneous bilateral internal pulmonary artery flow limiting device insertion. The PAB group mean age at banding was 141+/-54 days, and mean interval from PAB to transplant was 35+/ 31 days (range 1.5-68 days). No differences in age at transplant, weight at transplant, warm graft ischemia time or total graft ischemia time were detected between groups. Mean times of mechanical ventilation (control 143+/-69h vs. PAB 44+/-13h), inhaled nitric oxide (control 126+/-70h vs. PAB 37+/-9h), inotropic support (control 171+/-64h vs. PAB 87+/-17h), intensive care unit (ICU) stay (control 8.3+/-2.7 days vs. PAB 4.5+/-1.4 days), and hospital stay (control 10.4+/-3.9 days vs. PAB 7.0+/-1.1 days) were all lower in the PAB group (P<0.05 all comparisons). Two control patients died, three required extracorporeal membrane oxygenation (ECMO), and six did not tolerate primary chest closure. No PAB patient died or required ECMO. All PAB patients tolerated primary chest closure. All PAB patients had widely patent branch pulmonary arteries with no re interventions to date. All hospital survivors remain alive (mean follow-up, control 50.2 months, PAB 11.5 months). CONCLUSIONS: Pre-transplant mechanical limitation of pulmonary blood flow simplified management and reduced morbidity for HLHS patients undergoing heart transplantation at ages >or=4 months. This strategy extends the permissible transplant waiting time in older infants with HLHS. PMID- 12754027 TI - Outcomes in patients with low left ventricular ejection fraction after heart transplantation. AB - OBJECTIVES: Low left ventricular ejection fraction (EF) after heart transplantation (HT) is considered an ominous sign. We reviewed our database in order to determine outcomes in patients with low EF after HT and to identify a subset of patients who would benefit from immediate retransplantation. METHODS: We identified 825 patients who underwent HT at our institution between December 1983 and July 1999. Of these, 81 patients (70 men, 11 women; age, 48+/-12 years) had low (<35%) EF as determined by radionuclide ventriculography. Post transplantation survival; duration of low-EF episodes (>2 years vs. <2 years); and incidence of transplant rejection, infection, and transplant coronary artery disease (CAD) were determined for these patients. RESULTS: On average, low EF developed 800+/-1029 days after HT and lasted 550+/-756 days until improvement, repeat HT, or death of the patient. Actuarial survival was 79% at 1 year, 55% at 3 years, and 46% at 5 years. Shorter (<2-year) episodes of low EF tended to have an earlier onset than prolonged (>2-year) episodes (656 days vs. 1341 days) (P=0.014). Patients with prolonged episodes (n=17) survived longer than patients with shorter episodes (n=64) (2247 days vs. 1266 days) (P=0.002). The incidence of hemodynamically significant rejection was lower in the prolonged low-EF group (6% [1/17] vs. 26% [17/64]) (P=0.03). The incidence of infection (31% vs. 53%) and incidence of transplant CAD (47% vs. 39%) did not differ significantly between the prolonged and shorter low-EF groups. CONCLUSIONS: Low EF after HT, especially with later onset, is not associated with poor survival and is not related to hemodynamically significant rejection. These data further indicate that the presence of low EF even in the setting of CAD is not by itself an indication for repeat HT. PMID- 12754028 TI - The Jarvik 2000 is associated with less infections than the HeartMate left ventricular assist device. AB - OBJECTIVES: Device-related infections remain a considerable problem of left ventricular support. We compared the device-related-infections between the HeartMate left ventricular assist device (LVAD) and the Jarvik 2000 permanent LVAD, a device with a novel retroauricular power-supply. METHODS: Between December 2000 and September 2002 we implanted the HeartMate-vented, electrical system in 11 patients and the permanent Jarvik 2000 in six patients. Total support time was 1626 patient-days (HeartMate, 26-271 days) versus 1246 patient days (Jarvik 2000, 8-411 days). As potential risk factors for infection we analyzed age, preoperative hospital-days, total protein, cardiac index, maximal oxygen uptake, use of inotropes, LVAD risk-score-index and Aaronson-Mancini score, intubation time, and intensive care unit stay. We used the Center of Disease Control definitions for surgical site infections. RESULTS: HeartMate patients were younger than Jarvik 2000 patients (46+/-13 versus 58+/-6 years, P=0.056), there were no other differences in the risk factors. Four HeartMate patients needed late (>or=48 h) surgical revisions for bleeding/hematomas versus no revisions in the Jarvik 2000 patients. In the HeartMate-patients, there were seven (64%) driveline-infections, five (45%) device-pocket infections, and three (27%) bloodstream-infections, or 0.43 device-related infections/100 patient-days. Infections occurred early (34+/-31 days). Three patients required urgent transplantation due to bloodstream infection. There were no adverse outcomes in the HeartMate-group due to infection. In the Jarvik 2000 patients, there was one driveline-infection (16%) after 270 days of support (0.08 device-related infections/100 patient-days), significantly less than in the HeartMate-group (P=0.044). Driveline infections resolved with antibiotics and local wound care in the Jarvik 2000 patient, but only in one of seven HeartMate-patients. CONCLUSIONS: Implantation of the Jarvik 2000 is associated with less device related infections than the HeartMate-LVAD. The power-supply of the permanent Jarvik 2000 is suitable for long-term mechanical support. PMID- 12754029 TI - Determinants of long-term mortality of current palliative surgical treatment for dilated cardiomyopathy. AB - OBJECTIVE: Dynamic cardiomyoplasty and partial left ventriculectomy have shown limited and controversial results in the treatment of dilated cardiomyopathies. This study investigates causes and determinants of long-term mortality after these procedures. METHODS: Forty-three patients submitted to dynamic cardiomyoplasty and 43 who underwent partial ventriculectomy were studied. Patients were in New York Heart Association (NYHA) class III or IV before the procedures. In dynamic cardiomyoplasty group, hospital mortality was 2.2% and patients were followed for 48+/-31 months. Nine hospital deaths occurred after partial ventriculectomy and the remaining patients were followed for 38+/-29 months. RESULTS: For patients submitted to dynamic cardiomyoplasty, 1-year event free survival was 81.3+/-5.9%; 2-year, 65.1+/-7.2%; and 6-year, 23.1+/-6.7%. Partial left ventriculectomy patients presented event-free survival rates of 58.1+/-7.5%, 46.6+/-7.6% and 21.6+/-6.4% at the same periods, respectively. Late deaths were equally related to heart failure progression and arrhythmia events in both groups. Preoperative NYHA class IV, pulmonary hypertension and absence of left ventricular (LV) function improvement at the time of the final event were identified as independent predictors of poor long-term event-free survival and heart failure progression in cardiomyoplasty patients, while NYHA class IV, elevated serum nor-epinephrine and absence of LV function improvement were associated with these events after partial left ventriculectomy. Arrhythmia related deaths were only predicted by previous events of sustained ventricular tachycardia in partial left ventriculectomy group. CONCLUSIONS: Long-term results of dynamic cardiomyoplasty and partial left ventriculectomy are limited by patients' preoperative condition, by the loss of LV function benefits and by high incidence of sudden cardiac death. Palliative surgical treatment of dilated cardiomyopathies needs to be indicated earlier and may achieve better efficiency with the combination of different procedures to provide sustained improvement of LV function, to interrupt the progressive remodeling process and to prevent sudden cardiac death. PMID- 12754030 TI - Topical vancomycin applied on closure of the sternotomy wound does not prevent high levels of systemic vancomycin. AB - OBJECTIVE: Vancomycin is effective in reducing the risk of mediastinits and topical vancomycin has been hypothesised to give high local dose concentrations while avoiding high systemic levels, thus avoiding the risk of bacterial resistance to this second-line antibiotic. However, this theory has never been tested and the degree to which vancomycin is absorbed systemically is unknown. METHODS: Fourteen patients undergoing elective coronary artery bypass grafts (CABG) received 500mg of topical vancomycin prior to sternotomy closure. Serum samples were taken at 30, 60, 120, 180 and 720min post-operatively. In addition, samples were taken from the drain bottles and urine samples taken daily for 5 days. Vancomycin levels were measured by fluorescence polarisation immunoassay, using the reverse dilution method to give a detection limit of 0.8mg/l. RESULTS: Vancomycin was detected in almost all serum samples. Peak concentration was at 30min and the mean value was 2.96mg/l (range, 0.99-5.00mg/l). This mean fell to 1.32mg/l at 6h. Of the 500mg of vancomycin applied, a mean of only 8.8mg was found to have been lost into the drain bottles in the first 24h (range, 0.17 12.5mg). When 5 consecutive days of urine collection was achieved, a mean of 151mg of vancomycin was excreted (range, 40-195mg) and vancomycin was detectable in the urine till day 5. The mean concentration of vancomycin in the urine was maximal on day 1 and was 24.4mg/l (range, 4.49-44.98mg/l). CONCLUSIONS: Topical vancomycin causes significant systemic concentrations in the 6h post-surgery and can be detected in the urine for up to 5 days post-surgery. PMID- 12754031 TI - Total arch replacement using antegrade selective cerebral perfusion with right axillary artery perfusion. AB - OBJECTIVE: Right axillary artery (AxA) perfusion, which can prevent cerebral embolism caused by retrograde perfusion via the femoral artery (FA), was used for selective cerebral perfusion (SCP) as well as cardiopulmonary bypass (CPB) in aortic arch repair. We review the outcome of aortic arch surgery using SCP with right AxA perfusion to clarify its efficacy. METHOD: Between 1998 and 2002, 120 patients underwent aortic arch repair using SCP with right AxA perfusion. The mean age was 69+/-10 years. Aneurysms were atherosclerotic in 79, dissecting in 32, and others in nine patients. Twenty of them (16.7%) required emergency surgery. CPB was initiated with right AxA and FA perfusion, and following SCP was established using right AxA and left common carotid artery perfusion. RESULTS: With right AxA perfusion, hospital mortality was 5.8%. Multivariate analysis showed only ruptured aneurysm was an independent determinant for hospital mortality. Permanent neurological dysfunction developed in one patient (0.8%), while seven (5.8%) suffered from temporary one. In univariate analysis, SCP time, stenosis of the carotid arteries, past history of cerebrovascular events, and atherosclerotic aneurysm were not related to temporary neurological deficits CONCLUSION: Right AxA perfusion in conjunction with SCP is a safe and useful alternative for brain protection in total arch replacement. PMID- 12754032 TI - Technical advances of pulmonary thromboendarterectomy for chronic thromboembolic pulmonary hypertension. AB - OBJECTIVE: To minimize the side-effects of circulatory arrest times and profound hypothermia in patients undergoing pulmonary thromboendarterectomy (PTE) for chronic thromboembolic pulmonary hypertension (CTEPH). METHODS: Between March 2000 and June 2002, 30 patients (in New York Heart Association (NYHA) class III or IV) were operated for CTEPH using our modified technique. It includes moderate hypothermic (28-32 degrees C), total cardiopulmonary bypass (CPB) and simultaneous selective antegrade cerebral perfusion and occlusion of the bronchial arteries by introducing an occlusive balloon catheter into the descending aorta. The preoperative pulmonary vascular resistance in the cohort was 873+/-248dynes/s/cm(-5). RESULTS: Mean total CPB, cross-clamp times and duration of anterograde cerebral perfusion were 132+/-40, 98+/-21 and 21+/-10min, respectively. Mean core temperature 29.5+/-1.9 degrees C. The duration of postoperative mechanical ventilatory support was 34+/-44h and the mean stay in the ICU was 5+/-9 days. Seven patients had mild to moderate lung reperfusion injury, one transient neurological dysfunction. Three patients (10%) died during their hospital stay, two for multiorgan failure and one for persistent pulmonary hypertension. All patients had a significant pulmonary hemodynamic improvement and all achieved NYHA class I (P<0.01) status 4 weeks after discharge, remaining stable at a median follow-up time of 16 months (range, 1-29 months) postoperatively. CONCLUSIONS: These technical advances improve neurological outcome, control back-bleeding from bronchial arteries and avoid prolonged rewarming phases in patients undergoing PTE. PMID- 12754033 TI - Traumatic rupture of the innominate artery. AB - OBJECTIVE: Blunt traumatic rupture of the innominate artery is uncommon. We reviewed our experience to correlate the impact of patient stability, presence of associated injuries and location of the injury within the artery with outcome. METHODS: A retrospective review was performed of patients admitted between January 1, 1998 and December 17, 2002 with traumatic innominate artery rupture. Injuries were defined as proximal if they were 24h) 44% (P>or=0.002). Site of perforation, aetiology, and treatment strategy had no influence on mortality. The only independent predictor of mortality identified was time to diagnosis from perforation (beta 0.429, P=0.001). Time to definitive management in those undergoing an operative procedure had no influence on outcome with multivariate analysis. CONCLUSIONS: Prompt recognition of the diagnosis of oesophageal perforation and rapid institution of supportive measures, followed by an appropriate, patient specific treatment option optimises the chance of a successful outcome. The wide range of presentation of oesophageal perforation necessitates individualisation of treatment. PMID- 12754037 TI - The effect of age on the outcome of surgical treatment for carcinoma of the oesophagus and gastric cardia. AB - OBJECTIVE: The aim of this study was to examine the effect of age on the outcome of surgical treatment for carcinoma of the oesophagus and gastric cardia. METHODS: From 1979 to 1999, 596 patients underwent gastro-oesophagectomy with two field lymph node clearance for cancer under the care of a single surgeon. The clinicopathologic characteristics and survival of patients aged between 45 and 63 years (n=198, Group 1), 63 and 71 years (n=199, Group 2) and 71 and 89 years (n=199, Group 3) were compared. RESULTS: Thirty-day mortality for the first 300 patients (1979-1993) in this consecutive series was 5, 8 and 18% for Groups 1, 2 and 3, respectively, and 6, 6 and 6% for Groups 1, 2 and 3, respectively, in the second consecutive 296 patients (1993-1999, P=0.006, chi(2)). Tumours were poorly differentiated in 55.7, 59.1 and 53.4% of patients in Groups 1, 2 and 3, respectively, for 1979-1993 and 64.7, 53.2 and 40.2% of tumours in Groups 1, 2 and 3, respectively, for 1993-1999 (P=0.02, chi(2)). Adjuvant therapy was significantly more common in younger patients (P=0.006, chi(2)). Five-year survival in the first period was 22, 15 and 11% for Groups 1, 2 and 3, respectively, (P=0.02 log-rank) and 18, 16 and 14% for Groups 1, 2 and 3 in the second period (P=NS, log-rank). CONCLUSIONS: Elderly patients now have equivalent short and long-term outcomes compared to younger patients following gastro oesophagectomy. Five-year survival, even in younger patients receiving adjuvant therapy remains poor, however, at approximately 20%. New therapeutic modalities are required to improve long-term survival following surgical treatment of gastro oesophageal carcinoma. PMID- 12754038 TI - The impact of cardiovascular comorbidity on the outcome of surgery for stage I and II non-small-cell lung cancer. AB - OBJECTIVE: The association between lung malignancy and cardiovascular disease has been frequently reported though its therapeutic and prognostic implications not thoroughly analyzed. This study aims at assessing the possible impact of coexisting cardiovascular disease on the outcome of surgical treatment of non small cell lung cancer (NSCLC). METHODS: Among 247 consecutive patients undergone surgery for stage I and II NSCLC between 1990 and 1997, 34 (13.7%) had a cardiovascular comorbidity going to be treated by surgery, namely coronary artery disease (n=14), carotid stenosis (n=21), abdominal aortic aneurysm (n=9) and lower limbs arteriopathy (n=7). Among 22 patients (64.7%) who underwent cardiac/vascular surgery first, operation was performed after a median interval of 4.5 weeks. In five of this subset lung cancer was incidentally detected. In the other patients the cardiovascular disease was diagnosed and treated after the lung cancer had been detected and operated with a median interval of 3.5 months from thoracic procedure. Surgical procedures for lung cancer were three pneumonectomies, 12 lobectomies, 19 wedge resections. Uni and multivariate analysis for risk factors was carried out. RESULTS: In the group with cardiovascular comorbidity overall postoperative mortality was 9%, while morbidity rate was 58.8%, both of them primarily caused by cardiovascular disease and significantly higher for major resections. The 3- and 5-year survival rates were 54.8% and 35.5% compared to 69.2% and 56.4% among patients without cardiovascular comorbidity (P=0.01) while the timing of vascular surgery (before or after thoracic procedure) did not significantly affect survival. Multifocal vascular disease resulted the only positive factor at multivariate analysis (P=0.005, Odd Ratio=3.51, 95% Confidence Interval=1.4-8.4). CONCLUSIONS: Cardiovascular disease seems to have significant impact on survival and morbidity in patients undergone surgery for lung cancer, especially in presence of multifocal vascular disease and following major resections. The timing of vascular surgery and the extension of resection should rely on the severity of vascular disease, anaesthesiologist's and surgeon's final evaluation. PMID- 12754039 TI - Bronchioloalveolar lung cancer: occurrence, surgical treatment and survival. AB - OBJECTIVE: The prevalence of pulmonary adenocarcinoma has risen worldwide. Bronchioloalveolar carcinoma (BAC) was studied with regard to whether it exhibits a similar tendency, and its typical features were analysed. METHODS: Between 1992 and 2001, 278 lung resections were carried out for adenocarcinoma. Of these, 67 (24.1%) proved to involve BAC. Whereas BAC accounted for 6.9% of the cases in 1992, in 2001 the proportion was 46.9%. There were 37 men (55.2%) and 30 women (44.7%); the average age was 60.5 years. 58.2% of them had no complaints. Of the 26 non-smokers, 69.2% were women; of the 41 smokers, 29.2% were women. In consequence of the tumour, 49 lobectomies, three bilobectomies, six pneumonectomies and nine wedge resections were performed. RESULTS: The surgical mortality was 1.6%. The pathology revealed that 26 (38.8%) tumours were in stage I/A. In 15 cases (22.4%), tuberculosis (TB) could be revealed besides the BAC: by skin tests in four cases, by CT in three cases, by case history in four cases, and by pathology in four cases. For the overall group of 67 patients, the 5-year survival rate was 61.9%, and the mean survival time was 75.7 months. The 5-year survival rate among the women (74%) was significantly better than that among the men (37%) (P=0.030). There was no significant difference in survival with regard to the multiple BAC (85%). The 5-year survival rate was significantly worse in the mixed BAC group (20%) than in the non-mucinous (62.7%) and in mucinous (59%) group. The overall 5-year survival rate among the smokers and TB patients was 61 and 79%, respectively, which is higher than that among the non-smokers (47%) and non-TB patients (56%). The survival rate for the wedge resection cases was 37%, which was lower than that for the cases involving major resections (60%) (P=0.939). CONCLUSION: BAC has a favourable survival, particularly in women. In spite of this, resection smaller than lobectomy is recommended only as a compromise. A multiple appearance does not imply a worse survival. The best survival rate was found in the non-mucinous BAC among the histological groups. TB seems to be frequent among BAC patients. PMID- 12754040 TI - Bronchoscopic radioisotope injection for sentinel lymph-node mapping in potentially resectable non-small-cell lung cancer. AB - OBJECTIVE: Prospective study to evaluate the feasibility of a preoperative bronchoscopic radioisotope application, followed by conventional sentinel lymph node (SLN) identification and to investigate the occurrence and distribution of micrometastases in relation to SLN activity. METHODS: Twenty patients with a mean age of 63 years and proven clinical stage T1-3 N0-1 non-small-cell lung cancer (NSCLC) were included. A dosage of 80MBq radiolabeled technetium-99m nanocolloid was endoscopically administrated on intubated patients in the operation theatre. At thoracotomy, scintigraphic readings of both the primary tumor and hilar and mediastinal lymph-node stations were obtained with a hand-held gamma-counter. Patients underwent lung resection and mediastinal lymphadenectomy. Radiolabeled nodes were also examined separately on back-table. SLNs were defined as the hottest nodes or nodes with at least one-tenth of the radioactivity of the hottest nodes. SLNs pathologic assessment included standard examination using hematoxylin and eosin staining on step sections and immunohistochemistry (ICH) for cytokeratins. RESULTS: Identification of SLNs was possible in 19/20 (95%) patients after bronchoscopic radioisotope application. In 7/19 (37%) patients, a unique SLN was identified, whereas in 12/19 (63%) patients, nodes from two different stations could be classified as SLNs. Metastatic nodal disease was found in 9/19 (47%) patients. ICH revealed micrometastases in 2/12 (17%) patients, initially classified nodal negative. Pathologic negative SLNs were a predictor for absence of metastatic nodal disease after mediastinal lymphadenectomy. No complication related to the procedure was observed. CONCLUSION: Our preliminary results suggest that preoperative bronchoscopic radioisotope injection for SLN identification is a safe and simple method, improving accuracy of SLN detection in comparison to intraoperative technique. The absence of metastases in the SLNs seems to predict a negative nodal status accurately. PMID- 12754041 TI - Management for chest wall implantation of non-small cell lung cancer after fine needle aspiration biopsy. AB - OBJECTIVE: The implantation of cancer cells in the chest wall after percutaneous needle biopsy of the lung is rare. We investigated the clinical outcomes of implantation metastasis after percutaneous fine-needle aspiration biopsy of pulmonary mass suggestive of lung cancer. METHODS: Between January 1990 and December 2001, nine patients were treated for implantation metastasis of the chest wall. We retrospectively reviewed the patients' records and analyzed their clinical outcomes. RESULTS: During an 11-year period, 4365 patients underwent percutaneous fine-needle aspiration biopsy for indeterminate pulmonary nodule at Seoul National University Hospital. Eight patients developed implantation metastasis related to the procedure. One patient was presented to us after being biopsied in another hospital. A wide, full-thickness excision of the chest wall was performed in eight patients. In one patient, palliative chemotherapy was performed due to the presence of distant metastases in addition to the local recurrence. In six patients, postoperative adjuvant radiation was given. There was no surgical mortality or morbidity. The median survival was 96.5 months (range, 15-128 months) after pulmonary resection and 75 months (range, 8-93 months) after chest-wall resection. Six patients developed recurrence of the primary cancer in a median of 52 months (range 5-93 months). Three patients recurred at the chest-wall excision site and a wide, full-thickness chest-wall re resection was performed for two patients who recurred only at the previous chest wall excision site. Four patients are alive, four have died of recurrent disease, and one died of underlying lung disease. None died of implantation metastasis per se. CONCLUSIONS: The incidence of chest-wall implantation metastasis after fine needle aspiration biopsy is extremely rare. With successful resection, the prognosis for the patient seems to depend on the primary cancer. A radical and wide resection in conjunction with irradiation may provide long-term survival in patients with an initial early stage cancer. PMID- 12754042 TI - Long-term outcomes and risk factor analysis after pneumonectomy for active and sequela forms of pulmonary tuberculosis. AB - OBJECTIVE: The prevalence of pulmonary tuberculosis remains high in several areas of the world, and pneumonectomy is often necessary to treat the disease. We retrospectively analyzed the morbidities, mortalities, and long-term outcomes after pneumonectomy for the treatment of active tuberculosis or its sequelae. MATERIALS AND METHODS: Between 1981 and 2001, 94 patients underwent either pneumonectomy or pleuropneumonectomy for the treatment of tuberculosis. The patients included 44 males and 50 females and the mean age was 40 (16-68) years. The pathology included destroyed lung in 80, main bronchus stenosis in ten, and both lesions in four. Surgical procedures performed were pneumonectomy in 47, pleuropneumonectomy in 43, and completion pneumonectomy in four. RESULTS: One patient died postoperatively due to empyema. Twenty-three complications occurred in 20 patients: empyema in 15 (including seven bronchopleural fistulae), wound infections in five, and other complications in three. Univariate analysis revealed the presence of empyema, pleuropneumonectomy, prolonged operation time, old age, and intraoperative contamination as risk factors of postpneumonectomy empyema; it also showed that low preoperative FEV(1) and postoperative persistent positive sputum AFB were risk factors of bronchopleural fistula. In multivariate analysis, old age and low preoperative FEV(1) were risk factors of empyema while low preoperative FEV(1), positive sputum acid-fast bacilli, and the presence of aspergilloma were risk factors of bronchopleural fistula. There were 12 late deaths. Actuarial 5- and 10-year survival rates were 94+/-3% and 87+/-4%, respectively. CONCLUSION: Pneumonectomy could be performed with acceptable mortality and morbidity, and could achieve satisfactory long-term survival for the treatment of tuberculosis. In patients with risk factors, special care is recommended to prevent postoperative empyema or bronchopleural fistula. PMID- 12754043 TI - Huge mediastinal goiter. PMID- 12754044 TI - Nail gun penetrating cardiac injury. PMID- 12754045 TI - Axillary artery and transapical aortic cannulation as an alternative to femoral artery cannulation. AB - We present an experience with axillary artery and transapical aortic cannulation for cardiopulmonary bypass according to our indication. We could simply achieve antegrade flow using the two methods with satisfactory result. PMID- 12754046 TI - Gerbode's defect resulting from infective endocarditis. AB - We present a report of a Gerbode's defect (left ventricular-right atrial communication) resulting from bacterial endocarditis in a 63-year-old man. Also presented is a brief overview of the literature and a possible preoperative echocardiographic diagnostic criterion relating to this unusual condition. PMID- 12754047 TI - Systemic arterial supply to the left basal segment without the pulmonary artery: four consecutive cases. AB - Systemic arterial supply from the descending thoracic aorta to the basal segment of the left lower lobe without a pulmonary artery supply is a rare congenital anomaly within the spectrum of pulmonary sequestration cases. We encountered four consecutive cases, which were treated successfully by three basalectomies and one lower lobectomy to preserve lung function. PMID- 12754048 TI - Intimo-intimal intussusception: a rare clinical form of aortic dissection. PMID- 12754050 TI - Creation of a dual-coronary system for anomalous origin of the left coronary artery from the pulmonary artery utilizing the trapdoor flap technique. PMID- 12754053 TI - PTH2 receptor-mediated inhibitory effect of parathyroid hormone and TIP39 on cell proliferation. AB - The parathyroid hormone (PTH) has dual mitogenic and inhibitory effects on cell proliferation, depending on the cell type and experimental conditions. PTH can signal via two different receptors, both positively coupled to the adenylyl cyclase/cyclic AMP pathway which can mimic some of the proliferative effects of PTH. We evaluated the role of the type-2 PTH (PTH2) receptor on cell proliferation in clonal human embryonic kidney HEK293 cells stably expressing the human PTH2 receptor. Using a cyclic AMP-responsive gene-reporter, we confirmed that the tuberoinfundibular peptide (TIP39) and various human (h) PTH fragments including hPTH-(1-34) were potent agonists (EC(50) in the range of 0.01-0.3 nM) whereas the bovine (b) PTH peptides b(Tyr(34))PTH-(7-34) and its tryptophan derivative b[D-Trp(12),Tyr(34)]PTH-(7-34) behaved as antagonists (IC(50)=117 and 249 nM, respectively). hPTH-(1-34) produced a dose-dependent inhibition of cell proliferation (EC(50)=8.5+/-0.4 nM) after 3 days and this effect was fully reversed by the tryptophan derivative antagonist. The same effect was observed with TIP39 which caused a 30% maximal inhibition. These findings reveal that PTH2 receptor activation can inhibit cell proliferation and might explain the dual functionality of PTH on cell proliferation. PMID- 12754055 TI - Pharmacological properties of peptides derived from an antibody against the tachykinin NK1 receptor for the neuropeptide substance P. AB - Two peptides were derived from the structural analysis of a previously described monoclonal antibody [Mol. Immunol. 37 (2000) 423] against the tachykinin NK(1) receptor for the neuropeptide substance P. Here we show that these two peptides were able to inhibit the inositol phosphate transduction pathway triggered both by substance P and neurokinin A, another high-affinity endogenous ligand for the tachykinin NK(1) receptor. They also reduced the cAMP production induced by substance P. By contrast, only one antagonist peptide was able to prevent substance P and neurokinin A from binding the receptor, as revealed both by biochemical and autoradiographic studies. First, these results illustrate the generality of the antibody-based strategy for developing new bioactive peptides. Second, they indicate that antagonists, even exhibiting very close amino acid composition, can interact with the tachykinin NK(1) receptor at different contact sites, some of them clearly distinct from the contact domains for endogenous agonists. PMID- 12754054 TI - Mechanisms of nitric oxide independent activation of soluble guanylyl cyclase. AB - The heterodimeric heme-protein soluble guanylyl cyclase (sGC) is the only proven receptor for nitric oxide (NO). Recently, two different types of NO-independent soluble guanylyl cyclase stimulators have been discovered. The heme-dependent stimulator 2-[1-[2-fluorophenyl)methyl]-1H-pyrazolo[3,4-b]pyridin-3-yl]-5(4 morpholinyl)-4,6-pyrimidinediamine (BAY 41-8543) stimulates the enzyme in a synergistic fashion when combined with NO, requires the presence of the heme group and can be blocked by the soluble guanylyl cyclase inhibitor 1H-(1,2,4) Oxadiazole-(4,3-a)-quinoxalin-1-one (ODQ). The heme-independent activator 4-[((4 carboxybutyl)[2-[(4-phenethylbenzol) oxy]phenethyl]amino)methyl[benzoic]acid (BAY 58-2667) activates soluble guanylyl cyclase even in the presence of ODQ or rendered heme-deficient. In the present study, BAY 41-8543, BAY 58-2667 and NO strongly increased V(max). Combination of BAY 58-2667 and NO increased V(max) in an additive manner, whereas the synergistic effect of BAY 41-8543 and NO on enzyme activation was reflected in an overadditive increase of V(max). ODQ potentiated V(max) of BAY 58-2667-stimulated soluble guanylyl cyclase. BAY 41 8543 prolonged the half-life of the nitrosyl-heme complex of NO-activated enzyme, an effect that was not observed with BAY 58-2667. These results show the different activation patterns of both compounds and demonstrate their value as tools to investigate the mechanisms that underlie soluble guanylyl cyclase activation. PMID- 12754056 TI - The alpha2A-adrenoceptor subtype is not involved in inflammatory hyperalgesia or morphine-induced antinociception. AB - The purpose of the present study was to investigate the role of the alpha(2A) adrenoceptor subtype in inflammatory hyperalgesia, and in adrenergic-mu-opioid interactions in acute pain and inflammatory hyperalgesia. Behavioral responses to mechanical and thermal stimuli were studied in alpha(2A)-adrenoceptor knockout mice and their wild-type controls. Thermal nociception was evaluated as paw withdrawal latencies to radiant heat applied to the hindpaws. Mechanical nociception was measured using von Frey monofilament applications to the hindpaws. Mechanical and thermal hyperalgesia, induced with intraplantar carrageenan (1 mg/40 microl) were compared in alpha(2A)-adrenoceptor knockout and wild-type mice. The effects of the systemically administered mu-opioid receptor agonist morphine (1-10 mg/kg) were evaluated on mechanical withdrawal responses under normal and inflammatory conditions in knockout and wild-type mice. Withdrawal responses to radiant heat and von Frey monofilaments were similar in alpha(2A)-adrenoceptor knockout and wild-type mice before and after the carrageenan-induced hindpaw inflammation. Also, the antinociceptive effects of morphine in mechanical nociceptive tests were similar before and after carrageenan-induced hindpaw inflammation. Our observations indicate that alpha(2A)-adrenoceptors are not tonically involved in the modulation of inflammation-induced mechanical and thermal hyperalgesia. In addition, alpha(2A) adrenoceptors do not appear to play an important role in mu-opioid receptor mediated antinociception or antihyperalgesia. PMID- 12754057 TI - Involvement of spinal tyrosine kinase in inflammatory and N-methyl-D-aspartate induced hyperalgesia in rats. AB - Phosphorylation of a subunit of N-methyl-D-aspartate (NMDA) receptor by protein tyrosine kinase (PTK) Src or Trk is known to enhance its channel activity. We examined whether a spinally administered selective PTK inhibitor, lavendustin A, which has high affinity for Src and Trk tyrosine kinases, could influence the development and maintenance of inflammatory hyperalgesia or NMDA-induced hyperalgesia. Inflammation was induced by injection of a mixture of carrageenan and kaolin into the tail base of rats. In another group of rats, hyperalgesia was induced by intrathecal administration of NMDA. Intrathecal administration of lavendustin A (1.0 microg) or NMDA receptor antagonist, (+)-5-methyl-10,11 dihydro-5H-dibenzo[a,d]cycloheptane-5,10-iminemaleate, MK-801 (3.0 microg) before injection of a mixture of carrageenan and kaolin or after the development of inflammation inhibited carrageenan-kaolin-induced mechanical hyperalgesia. Intrathecal injection of 1.0 microg NMDA produced thermal and mechanical hyperalgesia. Co-administration of 1.0 microg lavendustin A with NMDA significantly reduced the duration of spontaneous pain behaviour and inhibited NMDA-induced hyperalgesia. Lavendustin A itself did not cause any sedation, motor impairment or analgesia. Our results suggest that inhibition of PTK could be therapeutically effective as an analgesic in some NMDA receptor-mediated hyperalgesic states. PMID- 12754058 TI - Mechanisms responsible for the in vitro relaxation of ligustrazine on porcine left anterior descending coronary artery. AB - In this study, we have evaluated the underlying mechanisms responsible for the relaxation response of ligustrazine (2,3,5,6-tetra-methyl-pyrazine; 2,3,5,6-MP) and its structural analogues (2-methyl-pyrazine (2-MP); ethyl-pyrazine (EP); 2,3 di-methyl-pyrazine (2,3-MP); 2,5-di-methyl-pyrazine (2,5-MP); 2,6-di-methyl pyrazine (2,6-MP) and 2,3,5-tri-methyl-pyrazine (2,3,5-MP)) in porcine left anterior descending coronary artery (tertiary branch, O.D. 2,3,5-MP>EP>2,5-MP>/=2,6-MP>/=2,3-MP>2-MP. Besides, salbutamol and forskolin caused an endothelium-independent relaxation. The relaxation response of ligustrazine, salbutamol and forskolin was blunted in the presence of cis-N-(2 phenylcyclopentyl) azacyclotridec-1-en-2-amine (MDL 12330A) (10 microM, an adenylate cyclase inhibitor) and N-[2-((bromocinnamyl)amino)ethyl]-5-isoquinoline sulphonamide (H-89, a protein kinase A inhibitor, 3 microM). Patch-clamp, whole cell electrophysiological studies using single smooth muscle cells of the left anterior descending coronary artery revealed that ligustrazine (300 microM), salbutamol (30 microM) and forskolin (1 microM) inhibited the nifedipine sensitive L-type Ca(2+) channels, and the inhibitory effect was eradicated by MDL 12330A (10 microM) and H-89 (1 microM). However, neither the Ca(2+)-dependent K(+) channel nor the ATP-dependent K(+) channel was modified by ligustrazine (300 microM). In conclusion, our results indicate that ligustrazine-mediated left anterior descending coronary artery relaxation is due to the activation of adenylate cyclase/protein kinase A cascade and the subsequent inhibition of nifedipine-sensitive, voltage-dependent L-type Ca(2+) channels. However, opening of K(+) channels seems to play no role in mediating the relaxation effect of ligustrazine. PMID- 12754059 TI - Nephroprotective effects of the endothelin ET(A) receptor antagonist darusentan in salt-sensitive genetic hypertension. AB - We tested the effect of selective endothelin ET(A) receptor blockade on the development renal damage in the Sabra rat model of genetic salt-sensitivity. Animals from the salt-sensitive (SBH/y) and salt-resistant strains (SBN/y) were either salt-loaded with deoxycorticosterone acetate and salt (DOCA) or fed a normal diet. Additional salt-loaded groups were also treated with the selective ET(A) antagonist darusentan (DA). Salt-loading in SBH/y increased systolic blood pressure by 75 mm Hg and urinary albumin excretion 23-fold (P<0.0001). Darusentan attenuated the rise of systolic blood pressure (50%) and urinary albumin excretion (63%, P<0.01, respectively). Salt-loading in SBH/y was associated with significant increased osteopontin mRNA expression as well as glomerulosclerosis and tubulointerstitial damage in the kidney (P<0.05, respectively). This was either significantly reduced or normalized by darusentan (P<0.05, respectively). Thus, darusentan confers a significant renal protection in the Sabra model of salt-sensitive hypertension. PMID- 12754060 TI - Testosterone depletion contributes to cyclosporine-induced chronic impairment of acetylcholine renovascular relaxations. AB - The immunosuppressant drug cyclosporine causes nephrotoxicity mainly via alterations of renovascular reactivity. This study investigated whether this effect of cyclosporine is modulated by the male gonadal hormone testosterone. The endothelium-dependent and -independent relaxations evoked by acetylcholine and sodium nitroprusside, respectively, were evaluated in phenylephrine preconstricted isolated perfused kidneys obtained from sham-operated, castrated, and testosterone-replaced castrated (CAS+T) male rats in the absence and presence of cyclosporine. Compared with sham-operated values, short-term (10 days) castration or cyclosporine treatment caused significant and equivalent reductions in plasma testosterone levels and vasorelaxant responses to acetylcholine. Treatment of castrated rats with cyclosporine caused no further attenuation of acetylcholine relaxations. Testosterone replacement of castrated (CAS+T) or cyclosporine-treated castrated (CAS+CyA+T) rats restored plasma testosterone and acetylcholine relaxations to near-sham-operated levels. On the other hand, castration caused significant increases in nitroprusside relaxations versus no effect for cyclosporine. The relaxant responses to nitroprusside in castrated rats were restored to sham-operated levels after testosterone replacement. Plasma urea and creatinine were not affected by castration but were significantly increased by cyclosporine. These findings suggest that testosterone exerts directionally opposite modulatory effects on endothelium-dependent and independent renal relaxations. Further, the results demonstrate that testosterone depletion may contribute, at least partly, to the inhibitory effect of cyclosporine on renovascular endothelial function. These data are clinically important because endothelial dysfunction contributes to vascular abnormalities associating cyclosporine therapy. PMID- 12754061 TI - Ultrasonographic demonstration of retropharyngeal lymph nodes: preliminary report. AB - The aim of this study was to describe the results of ultrasonography of upper retropharyngeal lymph node (RPN) metastasis in patients with pharyngeal carcinomas. A total of 10 patients with metastatic RPN were examined using percutaneous ultrasound (US) with 3.5-MHz probes. Primary cancer sites were the nasopharynx in two patients, the oropharynx in three and the hypopharynx in five. Metastatic RPNs lay in the level of occipital bone in five patients, C1 in nine, and C2 in five. US images were compared with previously obtained computerized tomography (CT) images based on size and depth. In all of the 10 patients, metastatic RPNs were ultrasonographically demonstrated as hypoechoic masses. Nodal sizes ranged from 1.5 cm to 3.5 cm both in CT and in US. Depths of the RPN centers were from 3.5 cm to 7.0 cm in CT, and from 3.5 cm to 6.5 cm in US. Differences of sizes and depths between CT and US were from -0.5 cm to 0.5 cm and from 0.0 cm to 1.0 cm, respectively. RPNs that are 1.5 cm or more in size can be demonstrated with percutaneous US using CT guidance. This technique should be utilized for the purpose of monitoring in a radiation therapy setting. PMID- 12754062 TI - Sonographic evaluation of the size of Achilles tendon: the effect of exercise and dominance of the ankle. AB - This study was undertaken to measure and compare the thickness and cross sectional area of the Achilles tendon between frequent- and infrequent-exercise subjects, and between the dominant and nondominant ankles in an asymptomatic Chinese population. Interobserver variability in the measurement of the size of Achilles tendon was also evaluated. High-resolution ultrasound (US) examination of Achilles tendons was performed in 40 healthy subjects (20 who frequently exercised, had exercise at least 3 days per week and at least 2 h per session; and 20 who infrequently exercised); their age range was 19 to 25 years. The thickness and cross-sectional area of the Achilles tendons were measured in a transverse scan at the level of medial malleolus. For each subject, the Achilles tendons were measured by five operators to evaluate the interobserver variability in the measurements. The mean thickness and cross-sectional areas of the Achilles tendon in a healthy Chinese population are 5.23 mm(2) and 56.91 mm(2)(2), respectively. The mean thickness of the Achilles tendon of frequent-exercise subjects (dominant ankle 5.43 mm, nondominant ankle 5.38 mm) was significantly greater than that of infrequent-exercise subjects (dominant ankle 5.08 mm, nondominant ankle 5.04 mm) (p < 0.05). The cross-sectional area of the tendons was also larger in frequent-exercise subjects but, whereas a significant result was found in dominant ankles (frequent-exercise subjects 60.46 mm(2)(2), infrequent-exercise subjects 54.71 mm(2)(2)) (p < 0.05), this was not the case for the nondominant ankles (frequent-exercise subjects 57.09 mm(2)(2), infrequent exercise subjects 55.4 mm(2)(2)) (p > 0.05). In both frequent- and infrequent exercise subjects, there was no significant difference in the mean thickness and cross-sectional area of Achilles tendon between dominant and nondominant ankles (p > 0.05). There was a high reproducibility in the sonographic measurement of the thickness (68%) and cross-sectional area (81%) of Achilles tendons. Results suggested that exercise would cause increase in the thickness and cross-sectional area of Achilles tendon. Interobserver variability is not significant in the sonographic measurement of Achilles tendons. PMID- 12754063 TI - Effect of Levovist on splanchnic hemodynamics in cirrhotic patients. AB - This study was aimed to assess the effect of Levovist on Doppler parameters of splanchnic hemodynamics. A total of 12 patients with cirrhosis and 12 healthy subjects underwent Doppler ultrasound (US) examination of the portal vein and of the hepatic, splenic and superior mesenteric arteries before, 5 to 8 and 12 to 15 min after the start of an 8-min long IV infusion of 2.5 g of Levovist. Mean velocity and mean diameter were calculated for the portal vein. Resistance index was determined for the arteries. A significant increase of resistance index was observed in the hepatic (0.80 +/- 0.07 vs. 0.71 +/- 0.06; p < 0.01) and splenic arteries (0.72 +/- 0.06 vs. 0.64 +/- 0.06; p < 0.01) 5 to 8 min after contrast agent injection in patients with cirrhosis, but not in controls. Neither portal vein diameter nor portal flow mean velocity changed during the test in both controls and cirrhotic patients. This effect might be related to a selective trapping of microbubbles in the altered hepatic and splenic microvasculature in patients with cirrhosis rather than being artefactual. It might have implications on harmonic imaging US protocols designed to image the cirrhotic liver in the early arterial phase. PMID- 12754064 TI - Hemodynamic evaluation of normally functioning Sulzer Carbomedics prosthetic valves. AB - The Sulzer Carbomedics prosthetic heart valve (CP) is a commonly used mechanical valve in clinical practice. In the present study, we used conventional and color Doppler echocardiography to assess the hemodynamics of normally functioning CP in the aortic (n = 73) and mitral (n = 127) positions. Our findings demonstrate no significant correlation of Doppler-measured peak and mean pressure gradients and effective orifice area with implanted valve size and actual orifice areas, measured directly by the manufacturer for CPs in both the mitral and aortic positions. However, it is still useful to measure effective orifice area by Doppler because a value in the normal or nonstenotic range points to an unobstructed prosthesis in the aortic or mitral position, in the absence of poor left ventricular ejection fraction. A value in the stenotic range could mean a normally functioning or obstructed prosthesis and, therefore, may need further investigation, such as assessment of valve leaflet motion by transthoracic or transesophageal echocardiography or fluoroscopy. Valve regurgitation as evaluated by color Doppler flow mapping was mild in practically all CPs in the aortic position, and in the majority of CPs in the mitral position. PMID- 12754065 TI - Chronological age modifies the microscopic remodeling process in viable cardiac tissue after infarction. AB - To define the impact of age on microscopic structural remodeling after myocardial infarction, the physical properties of infarct scar tissue and viable remote zone tissues in young (3 months) and older adult (18 months) Fischer rats were quantified with the use of high-frequency (50 MHz) high-resolution acoustic microscopy 3 months after coronary artery occlusion. We observed that integrated backscatter increased by 100% in the viable zones of old animals after infarction, but remained relatively unaffected in the same regions of younger animals. Mathematical models of myocardial scattering behavior indicated that a 25% increase in stiffness of the extracellular matrix materials in viable zones likely occurred in the older animals. Alterations in gross tissue collagen content were not responsible for this increased stiffness. These observations are compatible with the hypothesis that progressive age-related changes in the quality of the collagen (e.g., excessive age-related crosslinking) rather than its amount per se may have altered the stiffness of the extracellular matrix of remodeled viable tissue in older animals. PMID- 12754066 TI - Contrast-specific ultrasonic flow measurements based on both input and output time intensities. AB - Ultrasonic contrast agents are used to assess perfusion conditions based on evaluation of the time-intensity curve. Such a curve reflects the concentration of microbubbles in the perfused area and the indicator-dilution theory is used to derive the volumetric flow rate from the measured concentration. Previous results have shown that the technique is not reliable in some conditions due to the shadowing effect. To overcome this problem, a contrast-specific technique using both the input and output time-intensity relationships is proposed; this contrasts with conventional techniques that utilize only the relationship directly from the perfused area. The proposed technique is referred to as the input-output time-intensity curve (IOTIC) method. In this work, the shadowing effect was studied experimentally and the efficacy of the IOTIC technique was assessed and compared with conventional techniques. The results indicate that the IOTIC technique eliminates the shadowing effect and provides a good correlation between the actual flow rate and measured flow-related parameters; thus, making quantitative estimation of perfusion feasible. Note that the IOTIC is applicable, based on the assumption that both the input and the output can be positioned within the same image plane; its clinical applications include situations where the perfused area cannot be effectively imaged by ultrasound (US). One example is the assessment of brain perfusion, and it will be used as a target clinical application of the IOTIC technique. PMID- 12754067 TI - Improvement in breast tumor discrimination by support vector machines and speckle emphasis texture analysis. AB - Recent statistics show that breast cancer is a major cause of death among women in developed countries. Hence, finding an accurate and effective diagnostic method is very important. In this paper, we propose a high precision computer aided diagnosis (CAD) system for sonography. We utilize a support vector machine (SVM) to classify breast tumors according to their texture information surrounding speckle pixels. We test our system with 250 pathologically-proven breast tumors including 140 benign and 110 malignant ones. Also we compare the diagnostic performances of three texture features, i.e., speckle-emphasis texture feature, nonspeckle-emphasis texture feature and conventional all pixels texture feature, applied to breast sonography using SVM. In our experiment, the accuracy of SVM with speckle information for classifying malignancies is 93.2% (233/250), the sensitivity is 95.45% (105/110), the specificity is 91.43% (128/140), the positive predictive value is 89.74% (105/117) and the negative predictive value is 96.24% (128/133). Based on the experimental results, speckle phenomenon is a useful tool to be used in computer-aided diagnosis; its performance is better than those of the other two features. Speckle phenomenon, which is considered as noise in sonography, can intrude into judgments of a physician using naked eyes but it is another story for application in a computer-aided diagnosis algorithm. PMID- 12754068 TI - The behaviour of individual contrast agent microbubbles. AB - In recent years, our knowledge of the behaviour of ultrasonic microbubble contrast agents has improved substantially through in vitro experiments. However, there has been a tendency to use high concentrations of contrast agents in suspension, so that ultrasonic backscatter data are generated by a cloud of microbubbles. Such experiments involve a variety of assumptions with validity that is open to question. In addition, high concentrations of microbubbles cannot be used to understand the behaviour of individual microbubble scatterers. This paper proposes a technique that minimises the number of assumptions that need to be made to interpret in vitro experimental data. The basis of the technique is a dilute suspension of microbubbles that makes single scattering events distinguishable. A commercial scanner was used to collect radio frequency (RF) data from suspensions of two different contrast agents, Quantison and Definity. Backscatter data were collected over a range of acoustic pressures. It was found that Definity provided a constant number of scattering events per unit volume of suspension for almost all applied acoustic pressures. Quantison demonstrated an increasing number of scattering events per unit volume with increasing acoustic pressure. Below 0.6 MPa, Quantison scatterers were not individually detectable and provided levels of backscatter similar to those of a blood-mimicking fluid, which suggests that Quantison microbubbles had almost linear scattering behaviour. At acoustic pressures greater than 0.6 MPa, both agents appeared to provide echoes from free bubbles. The change in the number of scatterers per unit volume with acoustic pressure cannot be demonstrated using high concentrations of contrast agent. PMID- 12754069 TI - Lateral resolution in elastography. AB - The factors that control the lateral resolution in elastography were investigated using a simulation study. The lateral resolution was estimated from the simulated axial strain elastograms as the smallest measurable distance between two equally stiff lesions embedded in a homogeneously softer background. The lesions were symmetrically positioned lateral to the center of the target, at the focus of the transducer. Ultrasound (US) systems with different transducer frequencies, bandwidths and f-numbers were simulated. The effects of the ultrasonic parameters, the lateral spacing between adjacent echo signals, the cross correlation window length, the lesion/background elastic contrast and the lateral motion of scatterers on the estimated lateral resolution were investigated. The results show that the lateral resolution in elastography is proportional to the beam width of the US system used to acquire the data, and is on the same order as the sonographic lateral resolution. PMID- 12754070 TI - Hit/Miss monitoring of ESWL by spectral Doppler ultrasound. AB - The objective of this study was to investigate spectral Doppler ultrasound (US) for monitoring extracorporeal shock-wave lithotripsy (ESWL). In vitro experiments with model stones showed that Doppler spectra acquired after a shock wave hit result in a high peak followed by a decaying signal. The duration of decay was dependent on shock-wave energy, stone size, gas content of the water and the level of disintegration. It typically ranged from 30 ms to 150 ms. It was found, by comparison with optical high-speed imaging and US B-scan imaging, that the signal originated from fragments released by the stone and cavitation. If the monitored volume contained no target, the signal duration was significantly shorter. By this means, hits were reliably distinguished from misses. The results of clinical treatments were highly consistent with those of in vitro experiments. Therefore, spectral Doppler US is an excellent tool for hit/miss monitoring in ESWL. PMID- 12754071 TI - Biological and environmental factors affecting ultrasound-induced hemolysis in vitro: medium tonicity. AB - Whole human anticoagulated blood in vitro underwent controlled plasma replacement with either isotonic (0.9%) or hypotonic (0.5%) saline to 1. restore the blood to its original volume (which resulted in different hematocrits) or 2. bring the blood to a singular hematocrit (40%). The hypotonic cell MCVs were, on average, considerably larger than their isotonic counterpart by a ratio of 1.4:1. The blood samples were then subjected to two tests, one of mechanical fragility, the other to ultrasound (US)-induced hemolysis. The US exposure metrics were: 1.0-MHz center frequency, 200-micros pulse duration, 20-ms interpulse interval, exposure durations of 10 to 30 s in the presence of Albunex, as a control on blood gas nucleation, and exposure vessel rotation at 200 rpm. In all instances, the hypotonic blood displayed higher levels of hemolysis than the corresponding isotonic treatment. The highest ratio of US-induced hemolysis for the hypotonic:isotonic regimens was 2.2. In some instances, the ratio was somewhat less but appeared to be related to differences in whole blood viscosities among the regimens or other factors. The data supported the a priori hypothesis that hypotonicity will result in an increased tension on the cell membrane and render it more susceptible to shear-induced hemolysis, including exposure to US under conditions known to foster the occurrence of inertial cavitation. There was no temperature increase during the insonations of the blood. PMID- 12754072 TI - Inertial cavitation dose and hemolysis produced in vitro with or without Optison. AB - Gas-based contrast agents (CAs) increase ultrasound (US)-induced bioeffects, presumably via an inertial cavitation (IC) mechanism. The relationship between IC dose (ICD) (cumulated root mean squared [RMS] broadband noise amplitude; frequency domain) and 1.1-MHz US-induced hemolysis in whole human blood was explored with Optison; the hypothesis was that hemolysis would correlate with ICD. Four experimental series were conducted, with variable: 1. peak negative acoustic pressure (P-), 2. Optison concentration, 3. pulse duration and 4. total exposure duration and Optison concentration. P- thresholds for hemolysis and ICD were approximately 0.5 MPa. ICD and hemolysis were detected at Optison concentrations >/= 0.01 V%, and with pulse durations as low as four or two cycles, respectively. Hemolysis and ICD evolved as functions of time and Optison concentration; final hemolysis and ICD values depended on initial Optison concentration, but initial rates of change did not. Within series, hemolysis was significantly correlated with ICD; across series, the correlation was significant at p < 0.001. PMID- 12754073 TI - The pulse length-dependence of inertial cavitation dose and hemolysis. AB - Gas-based ultrasound (US) contrast agents increase erythrocyte sonolysis, presumably via enhancing inertial cavitation (IC) activity. The amount of IC activity (IC "dose") and hemolysis generated by exposure to 1.15 MHz US were examined with different US pulse lengths, but with the same delivered acoustic energy, for Optison and Albunex. The hypotheses were that 1. at longer pulse lengths, IC would generate more bubbles that could nucleate additional IC activity; 2. if the interval between pulse pairs were short enough for the next pulse to hit derivative bubbles before their dissolution, more IC could be induced; and 3. hemolysis would be proportional to IC activity. Two types of studies were performed. In the first, bubble generation after each burst of IC activity was quantified using an active cavitation detector (ACD), for different pulse lengths (5, 10, 20, 30, 50, 100 or 200 cycles), but the same pressure level (3 MPa) and total "on" time (173.16 ms). Low concentrations of either Optison or Albunex were added into the tank with high-intensity and interrogating transducers orthogonal to each other. For pulse lengths > 100 cycles, and pulse repetition intervals < 5 ms, a "cascade" effect (explosive bubble generation) was observed. In the second, IC was measured by passive detection methods. IC dose and hemolysis were determined in whole blood samples at a pressure level (3 MPa) and interpulse interval (5 ms) that induced the "cascade" effect. Each blood sample was mixed with the same number of contrast microbubbles (Optison approximately 0.3 v/v % and Albunex approximately 0.5 v/v %), but exposed to different pulse lengths (5, 10, 20, 30, 50, 100 or 200 cycles). With Optison, up to 60% hemolysis was produced with long pulses (100 and 200 cycles), compared with < 10% with short pulses (5 and 10 cycles). Albunex generated considerably less IC activity and hemolysis. The r(2) value was 0.99 for the correlation between hemolysis and IC dose. High pulse-repetition frequency (PRF) (500 Hz) generated more hemolysis than the low PRF (200 Hz) at 3 MPa. All experimental results could be explained by the dissolution times of IC-generated bubbles. PMID- 12754074 TI - Study of a "biological focal region" of high-intensity focused ultrasound. AB - The aim of this study was to explore a law of energy deposition of high-intensity focused ultrasound (HIFU) in various tissues and the expression of such a law. A focused ultrasound (US) tumor therapeutic system was used to apply a focused US beam to tissues both in vivo and in vitro. The formation of individual ellipsoid shaped regions of coagulative necrosis has been observed. Results showed that the volume of the ellipsoid-shaped coagulative necrosis region was different from that of the acoustic focal region (AFR), both in vitro and in vivo. Acoustic intensities ranging from 7 x 10(3) W/cm(2) to 27.7 x 10(3) W/cm(2) and exposure times from 1 to 20 s gave volumes of ellipsoid-shaped coagulative necrosis of 0.2 to 2000 mm(3). Although the HIFU doses applied were identical, the volumes of individual ellipsoid-shaped coagulative necrotic regions varied with the structures of tissues, their functional status and the irradiation depths. Individual ellipsoid-shaped regions of coagulative necrosis induced by HIFU can be added to produce coagulative necrosis of an entire tumor. We define the individual ellipsoid-shaped coagulative necrosis produced by the US energy deposition of a single exposure as the "biological focal region" (BFR) of HIFU. This serves as the basic unit for HIFU ablation of tumors, and is plotted as a function of AFR, acoustic intensity, exposure time, irradiation depth, the tissue structure and its functional status. PMID- 12754075 TI - Time to threshold (TT), a safety parameter for heating by diagnostic ultrasound. AB - Activities in the standardisation of medical ultrasonic imaging and monitoring devices have been undertaken to ensure the safe application and to provide the user with tools to perform risk-to-benefit analysis. Internationally, agreement has not been reached to implement a system based on a physical quantity such as temperature rise. Therefore, recently, the IEC Standard 60601-2-37 has adopted the thermal index (TI) as a safety parameter. TI indicates conditions that are more likely than others to produce thermal effects. The models used to calculate TI are based on the equilibrium temperature rise for a stationary transducer. Ultrasound (US) is often applied during a short time at one place. To characterise the situation before equilibrium is reached, this study proposes a new parameter: TT (time to threshold). TT indicates the time after which a threshold temperature rise is exceeded. TT is calculated for some tissue models to illustrate the meaning of TT. TT indicates how long one piece of tissue can be insonated safely, provided the safe threshold is known. Using TT, higher intensities can be applied safely during a limited time. This paper illustrates the effects of short insonation times and the influence of the beam diameter. Questions that are critical to the introduction of TT as a safety parameter are discussed. PMID- 12754076 TI - The effect of echo contrast agent on Doppler velocity measurements. AB - The purpose of this investigation was to determine the effect of echo contrast agents on spectral Doppler velocity measurements. SH U 508A was administered by IV injection in 15 patients. The transmitral flow velocity was measured at the E- and A-wave peaks before the start and at the peak of the contrast effect. The Doppler velocity was determined from the Doppler video spectral display and from power spectral analysis of the audio Doppler signal. The Doppler signal intensity was also measured. The Doppler signal intensity increased 17.4 +/- 3.5 dB (p < 0.0001) following echo contrast injection. This was associated with a significant increase in the spectral peak velocity as determined from either the video display or audio analysis. (p < 0.0001). The velocity corresponding to the audio power peak frequency (the modal velocity) did not change significantly (p = NS) and was independent of Doppler signal strength. PMID- 12754077 TI - An analytic derivation for the transient temperature rise during an ultrasound pulse focused on bone. AB - An analytic derivation is given for the maximum transient temperature rise due to millisecond ultrasound (US) pulses focused on bone. The temperature rise is found to have, within a small correction factor, a square-root dependence on the pulse duration and is independent of the focal diameter. The equation developed is essentially the same as that found in a previous paper that obtained the formula by numerical methods and subsequent curve fitting. PMID- 12754078 TI - Sex and gender in Brain Research Bulletin. PMID- 12754079 TI - The categorisation of male and female laboratory animals in terms of "gender". PMID- 12754080 TI - Forebrain Fos responses to reproductively related chemosensory cues in aromatase knockout mice. AB - Sexually relevant pheromonal cues are detected by the vomeronasal system which includes the posterodorsal part of the medial amygdala, the posteromedial part of the bed nucleus of the stria terminalis and the medial preoptic area. Copulatory behavior is impaired in mice lacking functional aromatase, the enzyme converting testosterone into estradiol. In this study, we used male aromatase knockout (ArKO) mice to investigate the role of aromatase in the differentiation and activation of preference for male- or female-related odorants. Moreover, using Fos immunoreactivity as a marker of neuronal activation we investigated the ability of sex-related pheromonal cues to activate the vomeronasal system. Both gonadally intact wild-type and ArKO mice preferred to investigate urine from females. The lack of estrogens did not reverse odor preferences, i.e. male ArKO mice did not show a preference for male odors. Exposure to soiled bedding from females induced Fos-protein in the posterodorsal part of the medial amygdala, in the posteromedial part of the bed nucleus of the stria terminalis, and in the periventricular part of the medial preoptic area of both the genotypes. Exposure to soiled bedding from intact males induced Fos in the posterodorsal part of the medial amygdala in wild-type mice and in the periventricular medial preoptic area in wild-type and ArKO mice. These results suggest that preference for female related odors and the Fos-mediated activation of the vomeronasal system do not rely on estradiol. Furthermore, sensitivity to female chemosensory cues and copulatory behavior are uncoupled in this knockout model. PMID- 12754082 TI - Enhanced angiotensin-mediated responses in the nucleus tractus solitarii of spontaneously hypertensive rats. AB - Studies using an AT(1) receptor antagonist, losartan, demonstrated that depressor and bradycardic responses to angiotensin II (Ang II) injection into the nucleus tractus solitarii (NTS) are mediated via those receptors. We further characterized Ang II-evoked cardiovascular responses in this nucleus in spontaneously hypertensive rats (SHR) using a new, selective AT(1) receptor antagonist, valsartan. In alpha-chloralose-anesthetized Sprague-Dawley (S-D) rats, Wistar-Kyoto (WKY) rats, and SHR, unilateral injection of Ang II into the NTS decreased arterial pressure (AP) and heart rate (HR). This response was eliminated by preinjection of valsartan. Depressor responses were much greater in SHR than in WKY rats. In normotensive rats, bilateral valsartan injection did not alter baseline AP or HR, or baroreceptor reflex index (BRI) calculated as the maximal change in HR (bpm) divided by phenylephrine- or nitroprusside-induced maximal change in mean AP (mmHg). In SHR, this treatment did not alter baseline HR and BRI, but significantly increased AP. Preinjection of valsartan did not alter injected glutamate effects in any strain. Thus, stimulation of AT(1) receptors within the NTS contributes to cardiovascular regulation independently of the baroreceptor reflex and the glutamatergic system. This angiotensinergic system in SHR acts tonically to reduce AP. PMID- 12754081 TI - Effect of strain and sex on mu opioid receptor-mediated G-protein activation in rat brain. AB - Strain and sex differences in mu opioid-mediated antinociception have been reported in rodents. The present studies evaluated mu opioid receptor-mediated G protein activation in Lewis and Fischer 344 (F344) male and female rats using agonist-stimulated [35S]GTPgammaS binding. Compared to Lewis rats, F344 rats exhibited a 35% higher level of net DAMGO-stimulated [35S]GTPgammaS binding in striatum. Basal [35S]GTPgammaS binding was approximately 30% lower in thalamus of Lewis than F344 rats. Female Lewis rats also exhibited slightly ( approximately 15%) lower basal [35S]GTPgammaS binding in cingulate cortex relative to F344 rats of either sex. The relative efficacies of the mu partial agonists, morphine and buprenorphine, were also examined. Buprenorphine exhibited approximately 40% lower relative efficacy in the periaqueductal gray in Lewis compared to F344 rats, but no other relative efficacy differences were found between strains or sexes. Moreover, regional differences in the relative efficacy of buprenorphine were also detected in Lewis but not F344 rats. In contrast to these results, the only difference found between sexes was the 13% lower basal [35S]GTPgammaS binding in the cingulate cortex of female compared to male Lewis rats. These results suggest that differences in mu opioid receptor-mediated G-protein activation may contribute to strain differences in opioid antinociception, whereas sex differences may result predominantly from other mechanisms. PMID- 12754083 TI - Cell cycle protein expression in proliferating microglia and astrocytes following transient global cerebral ischemia in the rat. AB - Cerebral ischemia induces microglial and astroglial activation, which may play a crucial role in the development of ischemic neuronal damage. In this study, we examined the role of cell cycle proteins in glial proliferation in the hippocampus following 10min of global cerebral ischemia in the rat. Proliferating cells were identified with immunostaining for proliferating cell nuclear antigen (PCNA), and glial cells were visualized with immunostaining for microglial response factor-1 (microglia/macrophages) and glial fibrillary acidic protein (astrocytes). Expression of cyclin D1 and cyclin-dependent kinase-4 was also examined with double label immunohistochemistry. Proliferating cells in the CA1 region after ischemia consisted of microglia and much fewer astrocytes. Microglial activation and proliferation (7.6-fold increase in number after 7 days) were preceded by an increase in PCNA-positive microglia; 83% of microglia were PCNA-positive after 2 days. Astrocytes increased by 1.8-fold after 7 days, and only 6% of astrocytes became PCNA-positive by day 7. Cyclin D1 and cyclin dependent kinase-4 immunoreactivity appeared in these glial cells in parallel with the expression of PCNA. The findings suggest that the accumulation of brain macrophages elicited by transient cerebral ischemia is caused predominantly by activation and proliferation of resident microglia through the upregulation of cell cycle proteins. PMID- 12754084 TI - An exploratory factor analysis of the Tail Suspension Test in 12 inbred strains of mice and an F2 intercross. AB - To explore the genetic dimensions of the stress response in rodents, we tested 12 inbred strains of mice and an F2 intercross (n=745) on the Tail Suspension Test (TST) and the Tail Suspension-Induced Hyperthermia (TSIH) paradigm. These selected 12 strains provide a representative sampling of the genetic heterogeneity of mousedom. An F2 intercross was derived from NMRI and 129S6 strains, which differ in their responses on the TST. Both inbred strains and F2 mice underwent a standardized protocol of automated TST with two sessions: (1) baseline and (2) imipramine TST. The duration of immobility and the body temperature after TST were recorded. The inbred strains were also tested in the Light-Dark Transition (LDT) test and in the Open Field Test (OFT), measuring the distance traveled, vertical movements, and center time as independent variables. The F2 mice were measured for core temperature after TST (TSIH). High intercorrelations among strain means were found for the LDT and OFT measures. Principal components analysis extracted four factors: "exploratory fear," body weight, imipramine response on immobility, and "stress reactivity." These dimensions were largely confirmed in the F2 population with one additional factor: imipramine response on TSIH. The results support a distinction between "stress reactivity" as measured by the TST and "exploratory fear" behavior as measured by the LDT and OFT. PMID- 12754085 TI - Noradrenaline release in the median preoptic nucleus area caused by hemorrhage in the rat. AB - The present study was designed to examine whether noradrenergic projections from the A1 cell group in the ventrolateral medulla to the median preoptic nucleus (MnPO) transmit information from the peripheral baroreceptors. In urethane anesthetized male rats, extracellular concentrations of noradrenaline (NA) in the region of the MnPO in response to hemorrhage (5 or 10 ml/kg) were monitored with in vivo microdialysis methods. Hemorrhage significantly increased the NA release in the MnPO area. The enhancement of NA release in the MnPO area caused by hemorrhage was significantly attenuated by previous injections of the local anesthetic lidocaine (2 %, 0.2 microl), but not by saline (0.2 microl), into the A1 region. These results suggest that the noradrenergic projections from the A1 region are important for carrying the peripheral baroreceptor information to the MnPO. PMID- 12754086 TI - Distribution of immunoreactive GABA and glutamate receptors in the gustatory portion of the nucleus of the solitary tract in rat. AB - The distribution of glutamate (GLU) and gamma-aminobutyric acid (GABA) receptors within the gustatory portion of the rat nucleus of the solitary tract (gNST) was investigated using immunohistochemical, histological and neural tract tracing techniques. Numerous somata throughout the gNST were immunoreactive for alpha amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) and N-methyl-D-aspartate (NMDA) receptors, while few were labeled for kainate receptors. AMPA and NMDA receptors were particularly abundant in the rostral central (RC) subdivision of the gNST, which receives most of the primary afferent input from the oral cavity and contains most of the gNST neurons that project to the parabrachial nuclei (PBN). This finding supports electrophysiological evidence that AMPA and NMDA receptors are involved in responses to orosensory input and indicates that their action may influence ascending taste signals as well. Compared to the ionotropic GLU receptors, few cell bodies were immunoreactive for metabotropic GLU receptors. Somata immunoreactive for GABA(A) and GABA(B) receptors were located throughout the nucleus. The densest neuropil labeling was for GABA(A) receptors in the ventral (V) subnucleus, the gNST subdivision that sends output to brainstem oromotor centers. The distributions of immunolabeling for GLU and GABA receptors imply that different functional roles may exist for specific receptors within this nucleus. PMID- 12754088 TI - Role of the mesolimbic cholinergic projection to the septum in the production of 22 kHz alarm calls in rats. AB - The role of the ascending cholinergic projection from the laterodorsal tegmental nucleus (LDT) to septum in the production of 22 kHz ultrasonic vocalization was studied in adult rats, using behavioral-pharmacological and anatomical tracing methods. Direct application of carbachol, a muscarinic agonist, into the lateral septal region induced species-typical 22 kHz alarm calls. The septum receives cholinergic input from LDT, thus, activation with glutamate of predominantly cholinergic neurons of the LDT induced comparable 22 kHz alarm calls in the same animals. This glutamate-induced response from LDT was significantly reduced when the lateral septum was pretreated with scopolamine, a cholinergic antagonist. To investigate the localization of the cell groups projecting to septum, the fluorescent retrograde tracer, fluorogold, was pressure injected into the lateral septum and sections from these brains were also immunostained against choline acetyltransferase (ChAT) to visualize cholinergic cell bodies. Several ChAT fluorogold double-labeled cells within the boundaries of the LDT were found, while other fluorogold-labeled regions did not contain double-labeled cells. These results provide both direct and indirect evidence that at least a part of the mesolimbic ascending cholinergic projection from LDT to septum is involved in the initiation of the 22 kHz vocalization. It is concluded that the septum is an integral part of the medial cholinoceptive vocalization strip and the 22 kHz alarm vocalization is triggered from septum by the cholinergic input from the LDT. PMID- 12754087 TI - High potassium-induced activation of choline-acetyltransferase in human neocortex: implications and species differences. AB - The role of electrical and potassium (K(+))-induced depolarisation on choline acetyltransferase (ChAT) activity in human and mouse neocortical slices was studied. When [3H]-ACh release was evoked by two K(+) stimulations in human neocortex, the mean S(2)/S(1) ratio was significantly below unity. ChAT inhibitors, like bromo-acetylcholine and ocadaic acid, raised this ratio by 79 and 63%, respectively, suggesting that the diminished S(2)/S(1) value in the absence of ChAT inhibitors reflected an increased ChAT activity at S(2) following K(+) depolarisation at S(1). When stimulated electrically, however, the S(2)/S(1) ratio in human neocortex was near unity and ocadaic acid remained without effect. In parallel experiments on mouse neocortical slices, the S(2)/S(1) ratio was near unity in both electrically or K(+)-evoked [3H]-ACh release and was not altered by ChAT inhibition. ChAT activity following K(+) depolarisation was also determined directly. ChAT activation in human neocortical slices was highest at 10 and 20mM K(+). ChAT activity in mouse neocortical tissue was not altered by K(+) depolarisation. These results suggest that in human, but not in mouse, neocortex ChAT activity may be increased due to ongoing K(+) depolarisation. This increase of ChAT activity supports a cholinergic degeneration hypothesis which has been entitled "autocannibalism" by Wurtman [TINS 15 (1992) 177]. PMID- 12754089 TI - The algogenic-induced nociceptive flexion test in mice: studies on sensitivity of the test and stress on animals. AB - Recently we developed a new technique, known as peripheral nociception test or algogenic-induced nociceptive flexion (ANF) test, to study the in vivo signal transduction of pain at the peripheral nerve endings in mice. In the present report, we examined the sensitivity of the method to detect pain signal and the stresses induced by the test on experimental animals. In the algogenic-induced biting and licking (ABL) test, bradykinin could not induce significant biting licking response even at a dose of 1nmol. It induced significant biting-licking response only at 10nmol. However, with the ANF test, 100fmol of bradykinin was enough to produce sharp and significant nociceptive flexion response. Similarly, substance P, ATP and ONO-54918-07, a stable prostaglandin I(2) agonist, induced nociceptive flexion response in ANF test at much lower doses than needed to induce biting-licking responses in ABL test. Next, we measured the plasma corticosterone level after different nociception tests, which is a measure of stress on animals due to experimental manipulations. However, no significant rise in corticosterone level was observed with ANF test. Altogether, these findings indicate that the ANF test is a highly sensitive and less stressful technique to study in vivo mechanisms of pain at the peripheral nerve ending. PMID- 12754091 TI - Facilitation of drug entry into the CNS via transient permeation of blood brain barrier: laboratory and preliminary clinical evidence from bradykinin receptor agonist, Cereport. AB - One novel approach of transporting drugs into the central nervous system (CNS) involves the activation of receptors on the endothelial cells comprising the blood brain barrier (BBB). Recently the selective B(2) bradykinin receptor agonist, Cereport (also called RMP-7), has been shown to transiently increase permeability of the BBB. Although initially developed to increase the permeability of the vasculature feeding glioma, recent studies have demonstrated that Cereport also increases the delivery of pharmacological agents across the normal (i.e. nontumor) BBB. In this review paper, we discuss evidence of enhanced CNS delivery of carboplatin, loperamide, and cyclosporin-A, which are accompanied by enhanced chemotherapeutic, analgesic and neuroprotective effects, respectively. These observations suggest feasibility of Cereport as an adjunct therapy to pharmacological treatments that require drug availability in the CNS to exert therapeutic efficacy. Because many potential drugs for CNS disorders normally do not cross the BBB, Cereport-induced transient permeation of BBB stands as an efficacious strategy for enhancing pharmacotherapy. PMID- 12754090 TI - Presynaptic modulation of acetylcholine, noradrenaline, and serotonin release in the hippocampus of aged rats with various levels of memory impairments. AB - Aged (25-27 months) Long-Evans female rats were distinguished according to whether they showed no significant impairment (AU), moderate impairment (AMI), or severe impairment (ASI) in a spatial reference-memory task. Young (3-5 months) rats served as controls. Electrically evoked overflow of tritium was assessed in hippocampal slices preloaded with [3H]choline or [3H]serotonin (5-HT). Nicotine evoked overflow of tritium was measured after preloading with [3H]noradrenaline (NA). Choline acetyltransferase (ChAT) and acetylcholinesterase (AChE) activity, and concentration of monoamines were assessed in homogenates. Aged rats exhibited reduced accumulation of [3H]choline and [3H]5-HT, increased accumulation of [3H]NA, and weaker electrically evoked overflow of [3H]acetylcholine ([3H]ACh) and [3H]5-HT. The overflow of [3H]NA was not altered consistently by aging. Roughly, drugs acting presynaptically had comparable effects in aged rats: oxotremorine and CP 93,129 inhibited the overflow of [3H]ACh, CP 93,129 and UK 14,304 reduced that of [3H]5-HT. ChAT or AChE activity, and 5-HT concentration were not changed by age; NA concentration was reduced. When significant, changes were comparable in AU, AMI, and ASI rats. Data show that aging alters cholinergic and serotonergic hippocampal innervations, release of ACh and 5-HT, but not presynaptic release-modulating mechanisms. These alterations do not account for variability in water-maze performance of aged rats. PMID- 12754092 TI - Comparative effects of injecting 5,6-dihydroxytryptamine in the dorsal or medial raphe nuclei on rat puberty. AB - The role played by the serotoninergic system in the control of puberty onset and first ovulation in rats is studied in this paper by analyzing the effects of injecting the neurotoxin 5,6-dihydroxytryptamine (5,6-DHT) into the dorsal (DRN) or medial (MRN) raphe nucleus of 30-day-old female rats. Complete lesion to the DRN resulted in the blockade of ovulation and a decrease in both the number of ovarian follicles and the serum concentration of follicle stimulating hormone (FSH). This treatment was also found to be associated with an increase in serotoninergic activity in the anterior and medial hypothalami. A lesion to the central portion of the DRN resulted in a significant decrease in the concentration of progesterone in serum and in the number of ova shed by ovulating animals. The lesion to the lateral portion of the DRN did not have an apparent effect on ovulation rate, the number of ova shed, nor in hormone serum concentration. The injection of propranolol to rats with a lesion to the DRN restored ovulation in 73% of treated animals and returned serotoninergic activity in the anterior hypothalamus to levels similar to those of sham-operated animals. In turn, in the medial hypothalamus, the increase in serotoninergic activity was not modified. The results presented herein suggest that serotoninergic inputs to the anterior hypothalamus have a direct influence on gonadotropin secretion and first ovulation, while the noradrenergic innervation exerts an indirect influence. PMID- 12754093 TI - Evaluation of the probes 2',7'-dichlorofluorescin diacetate, luminol, and lucigenin as indicators of reactive species formation. AB - This study attempts to provide a critical assessment of three different common approaches to identifying teactive species formed in biological systems: the 2',7'-dichlorofluorescin diacetate (DCFH-DA) assay, and the luminol- and lucigenin-amplified chemiluminescence assays. There have been several contradictory reports about the specificity of these methods. Our results show that DCFH is oxidized to the fluorescent compound 2',7'-dichlorofluorescin (DCF) in human neutrophils exposed to the following compounds: Aroclor (A)1242, hydrogen peroxide (H(2)O(2)), nitric oxide (NO), and FeSO(4). Use of a cell-free DCFH system showed increased formation of DCF by peroxynitrite (ONOO(-)), horseradish peroxidase (HRP) alone, and HRP in combination with H(2)O(2), FeSO(4) alone, and a mixture of FeSO(4) and H(2)O(2). The hydroxyl radical (z.rad;OH) scavenger formate and the iron ion chelator deferoxamine reduced the DCF formation induced by FeSO(4) in combination with H(2)O(2). DCFH was insensitive to NO and H(2)O(2) in the cell-free system. In the presence of neutrophils, the A1242-induced luminol chemiluminescence was decreased by the superoxide dismutase inhibitor diethyldithiocarbamic acid (DDC) and the myeloperoxidase inhibitor salicylhydroxamic acid (SHA). Exposure of the neutrophils to NO, FeSO(4), or H(2)O(2) alone did not have any effect. A1242-induced lucigenin chemiluminescence in the neutrophils was increased slightly by DDC, but was not affected by SHA, NO, FeSO(4), or H(2)O(2). In conclusion, we suggest that the DCF assay is only suitable for measurements of ONOO(-), H(2)O(2) in combination with cellular peroxidases, and z.rad;OH. Luminol is sensitive towards HOCl, while lucigenin is oxidized by O(2)z.rad;(-). PMID- 12754094 TI - Effects of dehydroepiandrosterone on oleic acid accumulation in rat liver. AB - The purpose of the present study was to determine whether dehydroepiandrosterone (DHEA) affects de novo fatty acid synthesis, oleic acid formation, fatty acid oxidation, and very low density lipoprotein (VLDL) secretion, in relation to the accumulation of lipid containing oleic acid, in rat liver. The rates of hepatic de novo synthesis of both fatty acid and monounsaturated fatty acid, determined by incorporation of 3H from 3H(2)O into fatty acid, were increased markedly when rats were fed a diet containing 0.5% (w/w) DHEA for 14 days. The treatment of rats with DHEA also enhanced the conversion of [14C]stearic acid into oleic acid in the liver in vivo. DHEA did not suppress fatty acid degradation in the liver. Namely, mitochondrial palmitic acid oxidation in liver homogenates and isolated hepatocytes was increased approximately 1.9- and 5-fold, respectively, in DHEA treated rats. Peroxisomal palmitic acid oxidation in isolated hepatocytes from rats treated with DHEA, however, was not significantly different from that of the control, despite the fact that peroxisomal degradation of palmitic acid in the liver homogenates was increased markedly. The rate of hepatic VLDL secretion in DHEA-treated rats was decreased markedly. These results indicate that the elevation of the hepatic fatty acid content, especially oleic acid, by DHEA feeding is due to an increase in both de novo fatty acid synthesis and the formation of oleic acid and to a decrease in the rate of hepatic VLDL secretion. Mitochondrial and peroxisomal fatty acid degradation does not appear to play a significant role in the accumulation of hepatic lipids. PMID- 12754095 TI - N-Acetylcysteine enhances UV-mediated caspase-3 activation, fragmentation of E2F 4, and apoptosis in human C8161 melanoma: inhibition by ectopic Bcl-2 expression. AB - Redox imbalance due to oxidative stress or excessive antioxidant levels can alter apoptotic responses. Recently, antioxidants like N-acetylcysteine (NAC) were reported to inhibit H(2)O(2)-mediated necrotic cell death, although they were inactive against apoptosis induced by other agents like etoposide. NAC was also found to kill preferentially tumor cells compared to normal fibroblasts at 20 50mM, but these concentrations are lethal to normal splenocytes. We now demonstrate that 10mM NAC, a non-toxic concentration, can enhance the UV radiation-mediated apoptosis of human C8161 melanoma cells. Compared to treatment with UV radiation alone, combination treatment with NAC doubled the ratio of activated caspase-3 to pro-caspase-3 and produced greater fragmentation of the retinoblastoma protein and the E2F-4 transcription factor without affecting the E2F-1 protein. These effects of joint NAC-UV radiation treatment were counteracted by the overexpression of the bcl-2 gene. To our knowledge, this report is the first to: (i) demonstrate a synergy between DNA-damaging agents, like UV radiation, and antioxidants, like NAC, and (ii) show that a Bcl-2 inhibitable E2F-4 fragmentation occurs concurrently with caspase-3 activation and apoptosis. PMID- 12754096 TI - Evidence of covalent binding of the dietary flavonoid quercetin to DNA and protein in human intestinal and hepatic cells. AB - Quercetin-rich foods have the potential to prevent human disease. However, knowledge of its biological fate and mechanism of action is limited. This study extends previous observations of the oxidation of quercetin by peroxidases to quinone/quinone methide intermediates and, for the first time, demonstrates covalent binding of [14C]quercetin to macromolecules. This was first demonstrated using horseradish peroxidase and hydrogen peroxide with human liver microsomal protein to trap the intermediates. To extend this observation to the cellular level, human intestinal Caco-2 cells and hepatic Hep G2 cells were incubated for up to 2hr with [14C]quercetin, and cellular DNA and protein were isolated. The cellular uptake of [14C]quercetin was rapid, and the covalent binding of [14C]quercetin to DNA and protein was determined by liquid scintillation spectrometry after extensive purification. Both cell types demonstrated DNA binding with a maximum level of 5-15pmol/mg DNA. The level of covalent binding to protein was considerably higher in both cell types, 75-125pmol/mg protein. To determine potential specificity in the protein binding, Hep G2 cells were treated with [14C]quercetin, and the cell lysate was subjected to SDS-PAGE followed by staining and autoradiography. Several distinct radiolabeled protein bands did not correspond to the major Coomassie blue stained cellular proteins. We propose that this specific binding may mediate part of the antiproliferative and other cellular actions of quercetin. PMID- 12754097 TI - Resistance to phorbol 12-myristate 13-acetate-induced cell growth arrest in an HL60 cell line chronically exposed to a glutathione S-transferase pi inhibitor. AB - Glutathione S-transferase pi (GSTpi; EC 2.5.1.18) has been shown recently to be a regulator of mitogen-activated protein kinases (MAPK). We have developed, by chronic exposure of HL60 cells to increasing concentrations of a peptidomimetic GSTpi inhibitor TLK199, a 10-fold resistant cell line (HL60/TLK199). Among the cellular adaptations observed in this cell line was an increase in extracellular signal-regulated kinase (ERK) activity without modification of basal expression levels. Phorbol 12-myristate 13-acetate (PMA) induced monocyte/macrophage cytodifferentiation in both HL60 wild-type (WT) and HL60/TLK199 cells. In contrast, PMA induced a pronounced cell growth inhibition and G(0)/G(1) cell cycle arrest in HL60 WT cells, while this differentiating agent had only a mild effect on cell growth without G(0)/G(1) cell cycle arrest in HL60/TLK199. This effect was associated with a rapid and sustained activation of ERK (up to 6hr) in HL60 WT cells but only a transient induction of these kinases (between 30 and 60min) in HL60/TLK199. Furthermore, treatment of both cell lines with PMA in combination with the protein tyrosine phosphatase inhibitors sodium orthovanadate (OV) or 3,4-dephostatin (DPN) circumvented the resistance to cell growth arrest and potentiated differentiation in HL60/TLK199 but had no effect on HL60 WT cells. The circumvention of the resistance to PMA was associated with a sustained activation of ERK. These data suggest that chronic exposure of HL60 cells to TLK199 alters cellular ERK activation by PMA, which may contribute to the differential response of the WT and resistant cells to PMA. PMID- 12754098 TI - Integrity of extracellular loop 1 of the human cannabinoid receptor 1 is critical for high-affinity binding of the ligand CP 55,940 but not SR 141716A. AB - Like other G-protein coupled receptors with hydrophobic ligands, the human cannabinoid receptor 1 (CB1) is thought to bind its ligands within the transmembrane region of the receptor. However, for some of these receptors the extracellular loops (ECs) have also been shown to play a role in ligand recognition and selectivity. We have taken a mutagenesis approach to examine the role of the amino terminus, EC1, and EC3 of CB1 in ligand binding. Eight mutant receptors, each with a dipeptide insertion, were constructed, expressed, and evaluated for binding to the cannabinoid ligands (-)-cis-3[2-hydroxy-4-(1',1' dimethylheptyl)phenyl]-trans-4-(3-hydroxypropyl)cyclohexanol (CP 55,940) and N (piperidin-1-yl)-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3 carboxamide hydrochloride (SR 141716A). Mutants with insertions in the membrane distal region of the amino terminus or EC3 maintained affinity for both ligands. Those with insertions in the membrane proximal region of the amino terminus or EC1 exhibited a loss of affinity for CP 55,940 while retaining wild-type affinity for SR 141716A. Representative mutants were analyzed for agonist-induced inhibition of cyclic AMP accumulation, and the results indicated that G-protein coupling remained intact. Alanine substitution mutants were made to address whether it was the perturbation of the overall structure of the region or the displacement of particular side chains that was responsible for the loss of CP 55,940 binding. We conclude that a structurally intact EC1, but not the comparably short EC3, is essential for high-affinity CP 55,940 binding. PMID- 12754099 TI - Mechanisms involved in exogenous C2- and C6-ceramide-induced cancer cell toxicity. AB - Ceramides are important intracellular second messengers that play a role in the regulation of cell growth, differentiation, and programmed cell death. To determine whether ceramides can mediate the apoptosis of HCT116 and OVCAR-3 cancer cells, exogenous C2-, C6-, and C16-ceramides were used to mimic the endogenous lipid increase that follows a large variety of stresses. C2- and C6 ceramides (cell-permeable ceramide analogs), but not C16-ceramide, induced nuclear factor-kappaB (NF-kappaB) DNA-binding, caspase-3 activation, poly(ADP ribose) polymerase degradation, and mitochondrial cytochrome c release, indicating that apoptosis occurs through the caspase cascade and the mitochondrial pathway. No difference in survival was observed between control cells and cells expressing mutated IkappaBalpha and treated with the permeable ceramides. This suggests that, at least in these cell lines, stable NF-kappaB inhibition did not modify the ceramide-induced cytotoxicity pathway. C6-ceramide also induced a double block in G1 and G2, thus emptying the S phase. PMID- 12754100 TI - Dual effect of ebselen on mitochondrial permeability transition. AB - This study reports an investigation on the effect of the seleno-organic compound ebselen on rat liver mitochondria. We show that low concentrations of ebselen induced an increase in rat liver mitochondrial membrane permeability, resulting in swelling and loss of membrane potential. These effects were mediated by the opening of the permeability transition pore. They required Ca(2+), were independent of pyridine nucleotide oxidation, and involved the oxidation of thiol groups. Ebselen pore induction is apparently promoted by the glutathione peroxidase mimicking activity of the drug. Opposite effects, that is, inhibition of both pore opening and thiol oxidation, were observed when concentrations higher than 20 micro M were used. These data demonstrate that ebselen is able to modulate the opening of the permeability transition pore and that it might be a critical event for both the proapoptotic and cytoprotective activities of the drug. PMID- 12754101 TI - Interplay between transcriptional and post-transcriptional regulation of Cyp2a5 expression. AB - The cytochrome P450 (Cyp) 2a5 gene can be upregulated transcriptionally or by mRNA stabilization. The heterogeneous nuclear ribonucleoprotein (hnRNP) A1 interacting with the CYP2A5 mRNA has been shown to be a key post-transcriptional regulator of the Cyp2a5 gene. The aim of this study was to investigate if the transcriptional and post-transcriptional steps of Cyp2a5 expression are linked. This was done by modifying the transcription rate with transcriptional inducers (phenobarbital and cyclic AMP) and inhibitors (actinomycin D and 5,6-dichloro-1 beta-d-ribofuranosylbenzimidazole) and analyzing the effects upon post transcriptional events. We found that inhibition of transcription led to relocalization of hnRNP A1 from the nucleus to the cytoplasm, to its strongly increased binding to the cytoplasmic CYP2A5 mRNA and to CYP2A5 mRNA stabilization. In contrast, stimulated transcription resulted in increased binding of nuclear hnRNP A1 to the Cyp2a5 promoter, and overexpression of hnRNP A1 led to stimulated transcription of a Cyp2a5 promoter-driven luciferase recombinant. This strongly suggests that the transcriptional and post transcriptional stages of Cyp2a5 expression are interrelated and that the nucleocytoplasmic shuttling hnRNP A1 may coordinate these different steps. PMID- 12754102 TI - Quantitative analysis of the Ah receptor/cytochrome P450 CYP1B1/CYP1A1 signalling pathway. AB - Cytochrome P450 (CYP) drug metabolising enzymes CYP1A1 and CYP1B1 are regulated through the ligand-activated aryl hydrocarbon (Ah) receptor. Differential expression of CYP1A1 and CYP1B1 mRNA and protein has previously been reported in human tissues with the presence of the message often extrapolated to indicate the presence of protein. The aim of this study was to clarify these potentially misleading findings, by analysing components of the Ah receptor pathway (CYP1B1, CYP1A1, Ah receptor and ARNT) using a combination of quantitative real-time RT PCR and immunoblotting. Three human cell lines (MOG-G-CCM, MCF7 and HEPG2) known to differentially express CYP1A1 and CYP1B1 mRNA and protein were exposed to the Ah receptor agonist 3-MC, and basal and inducible levels of CYP1A1, CYP1B1, Ah receptor and ARNT were determined. The key finding of this study was the demonstration of equivalent levels of CYP1B1 mRNA in both the treated and untreated MOG-G-CCM cell lines, with expression of the corresponding CYP1B1 protein only after exposure to an Ah receptor agonist. This finding suggests that a post-transcriptional mechanism is involved in the regulation of CYP1B1. In addition, the expression pattern of CYP1B1 mRNA and protein in the MOG-G-CCM cells highlights this cell line as a potential model for studying CYP1B1 expression in human tissue. PMID- 12754104 TI - Enzymatic characterization of recombinant mouse retinal dehydrogenase type 1. AB - Retinal dehydrogenases (RALDHs) convert retinal into retinoic acids (RAs), which are important signaling molecules in embryogenesis and tissue differentiation. We expressed mouse RALDH type 1 (mRALDH1) in Escherichia coli and studied the kinetic properties of the recombinant enzyme for retinal substrates. Purified recombinant mRALDH1 catalyzed the oxidation of all-trans and 9-cis retinal but not 13-cis retinal, and exhibited two pH optimums, 7.8 and 9.4, for all-trans and 9-cis retinal substrates, respectively. The K(m) for all-trans retinal (11.6 micro M) was 3-fold higher than for 9-cis retinal (3.59 micro M). However, the conversion efficiencies of either all-trans or 9-cis retinal to the respective RAs were similar. MgCl(2) inhibited the oxidation of both all-trans and 9-cis retinal. Chloral hydrate and acetaldehyde competitively suppressed all-trans retinal oxidation with inhibition constants (K(i)) of 4.99 and 49.4 micro M, respectively. Retinol, on the other hand, blocked the reaction uncompetitively. These data extend the kinetic characterization of mRALDH1, provide insight into the possible role of this enzyme in the biogenesis of RAs, and should give useful information on the determination of amino acid residues that play crucial roles in the catalysis of all-trans and 9-cis retinal. PMID- 12754103 TI - N6-Substituted adenosine derivatives: selectivity, efficacy, and species differences at A3 adenosine receptors. AB - The activation of the human A(3) adenosine receptor (AR) by a wide range of N(6) substituted adenosine derivatives was studied in intact CHO cells stably expressing this receptor. Selectivity of binding at rat and human ARs was also determined. Among N(6)-alkyl substitutions, small N(6)-alkyl groups were associated with selectivity for human A(3)ARs vs. rat A(3)ARs, and multiple points of branching were associated with decreased hA(3)AR efficacy. N(6) Cycloalkyl-substituted adenosines were full (/=6 carbons) hA(3)AR agonists. N(6)-(endo-Norbornyl)adenosine 13 was the most selective for both rat and human A(1)ARs. Numerous N(6)-arylmethyl analogues, including substituted benzyl, tended to be more potent in binding to A(1) and A(3) vs. A(2A)ARs (with variable degrees of partial to full A(3)AR agonisms). A chloro substituent decreased the efficacy depending on its position on the benzyl ring. The A(3)AR affinity and efficacy of N(6)-arylethyl adenosines depended highly on stereochemistry, steric bulk, and ring constraints. Stereoselectivity of binding was demonstrated for N(6)-(R-1-phenylethyl)adenosine vs. N(6)-(S-1 phenylethyl)adenosine, as well as for the N(6)-(1-phenyl-2-pentyl)adenosine, at the rat, but not human A(3)AR. Interestingly, DPMA, a potent agonist for the A(2A)AR (K(i)=4nM), was demonstrated to be a moderately potent antagonist for the human A(3)AR (K(i)=106nM). N(6)-[(1S,2R)-2-Phenyl-1-cyclopropyl]adenosine 48 was 1100-fold more potent in binding to human (K(i)=0.63nM) than rat A(3)ARs. Dual acting A(1)/A(3) agonists (N(6)-3-chlorobenzyl- 29, N(6)-(S-1-phenylethyl)- 39, and 2-chloro-N(6)-(R-phenylisopropyl)adenosine 53) might be useful for cardioprotection. PMID- 12754105 TI - Inhibition of microtubule polymerization by 3-bromopropionylamino benzoylurea (JIMB01), a new cancericidal tubulin ligand. AB - 3-Bromopropionylamino benzoylurea (JIMB01) is a small molecular weight compound (MW 313) that has been synthesized in our laboratory. This compound showed antiproliferative activities in a panel of thirteen human tumor cell lines with IC(50) values in the range of 0.25 to 0.51 micro M for leukemia and lymphoma cell lines and 0.33 to 9.26 micro M for solid tumor cell lines. The primary action of JIMB01 is to inhibit microtubule polymerization but not depolymerization. A 4 micro M concentration of the compound caused a complete inhibition of microtubule assembly in a cell-free reaction. An increase in the number of human hepatocarcinoma cells blocked in the M-phase was detected 12hr after exposure to JIMB01. The kinase activity of cyclin B1, which is responsible for the G(2)/M transition, was increased accordingly. Bcl-2 phosphorylation became visible, in a western blot, within 6hr in hepatocarcinoma cells treated with JIMB01 at 0.8 micro M or higher. JIMB01-induced apoptosis in liver cancer cells was confirmed by morphological methods, flow cytometry, as well as DNA gel electrophoresis, which clearly demonstrated DNA degradation in the form of a multiple-unit DNA ladder. Furthermore, in vivo experiments using nude mice showed that intraperitoneal injection of JIMB01 at 15mg/kg (with seven injections at 4-day intervals) significantly inhibited the growth of a human hepatocarcinoma (BEL 7402) by 66% in tumor volume (P=0.01), at least compatible to the inhibition by vincristine (43% inhibition), indicating good bioavailability of the compound in the circulation. Side-effects of the compound were not observed, and the body weight of the treated mice remained stable during the 4-week treatment. Since JIMB01 is a small compound, targets a specific molecule in tumor cells, and has promising activity against human hepatocarcinoma in vivo, we believe JIMB01 merits consideration for further investigation. PMID- 12754106 TI - Deregulation of gene expression in fetal oocytes exposed to doxorubicin. AB - Doxorubicin is an effective anticancer drug but its use is limited due to its adverse side effects such as infertility and cardiomyopathy. Some possible mechanisms of the action of doxorubicin have been postulated, but the initial gene deregulation response has not been investigated. Fetal life stages are critical periods in mammalian oogenesis. This study analyzes gene expression alterations in mouse fetal oocytes exposed in vitro to this anticancer agent. cDNA libraries were generated from isolated fetal oocytes and differential screenings performed with cDNAs from in vitro doxorubicin-treated and -untreated oocytes. Differentially expressed genes were assessed by real-time RT-PCR to quantify the extent of their transcriptional control in doxorubicin-exposed oocytes. The results show that doxorubicin alters the expression of genes involved in the mitochondrial respiratory chain, intracellular transport and cell differentiation. Finally, the up-regulation of a differentially expressed gene (metaxin) mediated by its promoter was evaluated in a functional assay. When treated with doxorubicin, somatic cells transfected with a genetic construct including the promoter of metaxin and a reporter gene showed increases in expression similar to those observed in fetal oocytes. This demonstrates the direct effect of agent on the regulation of a specific gene. PMID- 12754107 TI - High resolution X-ray structure of potent anti-HIV pokeweed antiviral protein III. AB - Pokeweed antiviral protein III (PAP-III), a naturally occurring protein isolated from late summer leaves of the pokeweed plant (Phytolacca americana), has potent anti-HIV activity by an as yet undetermined molecular mechanism. PAP-III belongs to a family of ribosome-inactivating proteins that catalytically deadenylate ribosomal and viral RNA. The chemical modification of PAP-III by reductive methylation of its lysine residues significantly improved the crystal quality for X-ray diffraction studies. Trigonal crystals of the modified PAP-III, with unit cell parameters a=b=80.47A, c=76.21A, were obtained using 30% PEG400 as the precipitant. These crystals contained one enzyme molecule per asymmetric unit and diffracted up to 1.5A, when exposed to a synchrotron source. Here we report the X ray crystal structure of PAP-III at 1.6A resolution, which was solved by molecular replacement using the homology model of PAP-III as a search model. The fold typical of other ribosome-inactivating proteins is conserved, despite several differences on the surface and in the loop regions. Residues Tyr(69), Tyr(117), Glu(172), and Arg(175) are expected to define the active site of PAP III. Molecular modeling studies of the interactions of PAP-III and PAP-I with a single-stranded RNA heptamer predicted a more potent anti-HIV activity for PAP III due to its unique surface topology and more favorable charge distribution in its 20A-long RNA binding active center cleft. In accordance with the predictions of the modeling studies, PAP-III was more potent than PAP-I in depurinating HIV-1 RNA. PMID- 12754108 TI - Inflammation and inducible nitric oxide synthase have no effect on monoamine oxidase activity in glioma cells. AB - Heightened monoamine oxidase (MAO) and inducible nitric oxide synthase (iNOS) activity can contribute to oxidative stress, the formation of active neurotoxins, and associated neurodegenerative diseases of the brain. Although these enzymes co exist within astrocytes, there has been little research examining the correlation between the two during inflammation. In this study, C6 glioma cells were stimulated with lipopolysaccharide (LPS):Escherichia coli 0111:B4 (6 micro g/mL):rat interferon-gamma (IFN-gamma) (100U/mL). In LPS/IFN-gamma-treated cells, the MAO substrates dopamine (DA) and tyramine caused a concentration-dependent attenuation of NO(2)(-) and NO(3)(-). In contrast, treatment with an MAO-A inhibitor (clorgyline) or an MAO-B inhibitor ((-)-deprenyl) did not reverse these effects. MAO activity was inhibited effectively by clorgyline and deprenyl; however, neither MAO inhibitor had an effect on NO(2)(-) in stimulated cells. Inversely, increasing concentrations of LPS/IFN-gamma resulted in heightened iNOS protein expression and NO(2)(-); however, these events did not correlate with any distinctive change in MAO enzyme activity. Moreover, a selective iNOS inhibitor, N(6)-(1-iminoethyl)-L-lysine, in LPS/IFN-gamma-stimulated cells caused a concentration-dependent attenuation of NO(2)(-) with no effects on MAO activity or iNOS protein expression. The attenuating effects of DA on iNOS were blocked completely by ICI 118-551 [(+/-)-1-[2,3-(dihydro-7-methyl-1H-inden-4-yl)oxy]-3 [(1-methylethyl)amino]-2-butanol hydrochloride], indicating a role for the beta(2)-adrenergic receptor. In conclusion, these data indicate that activity or expression of iNOS does not influence MAO activity in activated rat glioma cells. Moreover, DA exerts an inhibitory effect on glial iNOS through a receptor mediated cascade. PMID- 12754110 TI - Interaction of dimercaptosuccinic acid (DMSA) with angiotensin II on calcium mobilization in vascular smooth muscle cells. AB - Dimercaptosuccinic acid (DMSA) was shown to lower blood pressure in rat models of arterial hypertension. Thus, there is evidence that-besides its chelating properties-DMSA has a direct vascular effect, e.g. through scavenging of reactive oxygen species (ROS). We speculated that, in addition, intracellular calcium mobilization may be involved in this action. Therefore, the present study examined the effects of DMSA on Ca(2+) mobilization in cultured vascular smooth muscle cells (VSMCs) from rat aorta. Intracellular free Ca(2+) concentration ([Ca(2+)](i)) was measured with fura-2 AM. In a first series of experiments DMSA, 10(-11) to 10(-6)M, induced an immediate dose-dependent up to 4-fold rise of [Ca(2+)](i) (P<0.001) which was almost completely blunted by the calcium channel blocker verapamil or the intracellular calcium release blocker TMB-8. In a second series of experiments, when VSMCs were exposed acutely to DMSA (10(-11) to 10( 6)M), the angiotensin (ANG) II (10(-8)M)-induced rise in [Ca(2+)](i) to 295+/ 40nM was attenuated at the average by 49% independent of the dose of DMSA. Preincubation of VSMCs with DMSA (10(-6)M) for 60min reduced basal [Ca(2+)](i) by 77% (P<0.001) and dose-dependently attenuated the ANG II (10(-8)M)-induced rise in [Ca(2+)](i) between 28 and 69% at concentrations between 10(-9) and 10(-5)M DMSA, respectively (P<0.05 and <0.01). In the presence of TMB-8, which attenuated the ANG II (10(-8)M)-induced rise in [Ca(2+)](i) by 66%, DMSA (10(-6)M) had no additional suppressive effect on [Ca(2+)](i). The results suggest that DMSA acutely raises [Ca(2+)](i) by stimulating transmembrane calcium influx via L-type calcium channels and by calcium release from intracellular stores followed by a decrease in [Ca(2+)](i) probably due to cellular calcium depletion. Thus, in addition to its action as scavenger of ROS, which in part mediate the vasoconstrictor response, e.g. to ANG II, DMSA may exert its hypotensive effect through decreasing total cell calcium, thereby attenuating the vasoconstrictor induced rise in [Ca(2+)](i) in VSMCs. PMID- 12754109 TI - Delivery of pharmacologically active dexamethasone into activated endothelial cells by dexamethasone-anti-E-selectin immunoconjugate. AB - To deliver selectively anti-inflammatory agents into activated endothelial cells, drug-targeting conjugates were developed. Dexamethasone (Dexa) was covalently linked to a monoclonal antibody specifically recognizing E-selectin, which is strongly upregulated in endothelial cells at inflammatory sites. In the present study, the pharmacological effects of this Dexa-mouse antihuman E-selectin antibody (H18/7) (Ab(hEsel)) conjugate were investigated and compared to the effects obtained by free Dexa in human umbilical vein endothelial cells. Flow cytometry and ELISA were performed to analyze the levels of cell adhesion molecules (ICAM-1 and VCAM-1) and secreted cytokines (IL-6 and IL-8). The studies were extended by analysis of a complex gene expression pattern, using a cDNA expression array containing 268 genes encoding human cytokines/cytokine receptors. Fifty genes and 28 genes were upregulated (ratio> or =2) upon incubation of human umbilical vein endothelial cells with TNFalpha for 6 and 24hr, respectively. This gene expression profile was markedly altered when cells were activated with TNFalpha in the presence of Dexa (100 nM) or Dexa-Ab(hEsel) conjugate (10 micro g/mL conjugate corresponding to 100 nM Dexa). Relative and competitive RT-PCR analysis verified downregulation of TNFalpha-mediated expression of CD40L and IL-8 by Dexa and Dexa-Ab(hEsel), respectively. These results indicated a successful internalization and processing of Dexa-Ab(hEsel) in activated endothelial cells, allowing the intracellularly delivered Dexa to exert its pleiotropic anti-inflammatory activity. PMID- 12754111 TI - Adrenergic receptors in the bovine mammary artery. AB - The response of isolated preparations of bovine mammary artery was investigated, with the aim of characterising further the adrenergic receptor subtypes present. Noradrenaline (NA) and the alpha(1) agonist phenylephrine gave sigmoidal dose response curves with pEC(50) values of 5.97+/-0.07 (N=34) and 6.21+/-0.32 (N=8), respectively. Stimulation of alpha(2) receptors with UK 14,304 produced a weak response with pEC(50) of 6.78+/-0.38 (N=7), and maximal contraction of 17.8+/ 9.9% relative to NA. A61603, an alpha(1A) agonist, gave a curve parallel to NA, but shifted to the left (pEC(50) of 6.98+/-0.19 (N=5)); this drug had an increased potency of 10-fold relative to NA, and 4-fold relative to phenylephrine. Schild analysis of curves obtained with the alpha(1) antagonist prazosin gave a pA(2) of 8.70+/-0.47 (N=6-9), whereas the alpha(2) antagonist yohimbine resulted in a pA(2) of 7.65+/-0.16 (N=4). The alpha(1A) receptor antagonists WB4101 and 5-methylurapidil gave pA(2) values of 9.39+/-0.69 (N=4) and 7.72+/-0.02 (N=2-3), respectively. The irreversible alpha(1B) inhibitor CEC reduced the pEC(50) from 5.39+/-0.12 to 4.31+/-0.18 (N=7) only at the highest dose used, and high doses of the alpha(1D) antagonist BMY 7378 produced a shift to the right at giving a pA(2) of 7.37+/-0.08 (N=3). These results suggest major involvement of the alpha(1B) adrenergic receptor subtype in contraction of the bovine mammary artery, which is similar to the human internal mammary artery. PMID- 12754113 TI - Role of the biomechanical property of the endplate in anterior cervical fusion. AB - OBJECTIVE: To study the distribution of the biomechanical properties in the cervical vertebrae with or without the endplate, so as to evaluate the importance of the endplate in cervical anterior fusion. METHODS: Indentation tests were performed at 20 standardized testing points chosen on each surface of the endplate with the impact adjusted perpendicular to the endplate surface using a hemispherical indenter 2 mm in diameter. The failure load and rigidity at each test site were determined from the load-displacement curves. Independent sample t test and factorial analyses were used to analyze the results. RESULTS: On either superior or inferior endplate surface, both the failure load and rigidity differed significantly between the endplate-intact and endplate-removed groups (P < 0.001). The posterior region of the superior endplate and the lateral regions of the inferior endplate were stronger and more rigid than any other region across the endplate surface in the endplate-intact group, while for enplate removed group, the posterior region of the superior endplate and the posterolateral regions of the inferior endplate were the stronger and more rigid. CONCLUSION: The endplate plays an important role in deciding the success of anterior cervical fusion, and due attention should be paid to the endplate in the intervertebral implant device designing and the surgical approach modification. PMID- 12754112 TI - TNF-alpha induces apoptosis of parietal cells. AB - Helicobacter pylori infection can be associated with chronic gastric inflammation and hypochlorhydria with increased levels of the proinflammatory cytokines. The current study investigated the effects of TNF-alpha on programmed death of gastric parietal cells. TNF-alpha induced apoptosis of parietal cells in isolated perfused rat stomachs at 10ng/mL. In isolated and highly enriched rat parietal cells, 10ng/mL TNF-alpha induced a 2.6-fold increase in the apoptotic rate. The 55kDa protein of TNFR-1 but not the 75kDa of TNFR-2 was detected by Western blot analysis. TNF-alpha-induced apoptosis of isolated parietal cells was inhibited by pretreatment with different NF-kappaB-inhibitors, nitric oxide synthase inhibitors and with antisense-oligodeoxynucleotides against the p65 subunit of NF kappaB. Investigation of downstream signaling pathways of apoptosis revealed that TNF-alpha induced the expression of iNOS, but failed to stimulate the activity of caspase 3. The TNF-alpha effect on gastric parietal cells may contribute to the atrophy and hypochlorhydria of the gastric mucosa observed during chronic H. pylori infection. PMID- 12754114 TI - [A new approach for exploring the essence of meridian]. AB - The paper includes a brief review and evaluation of the progress in the research of the essence of meridian in China in the last two decades, in which the author suggests that more effort at all levels involving multiple disciplines needs to be investigated in the pursuit of this issue after the failure of prolonged research in the fields of histological anatomy. Microcosmic investigation, in particular, should be pursued making full use of the available achievement in molecular cell biology, genome research, biophysics and biochemistry and so forth, so as to build a solid plateform for making a breakthrough. The author urges the importance of research in the information carrier that regulates the meridian physiological network of the body. Based on the theories in cellular signal transduction and relevant experimental data, the author points out that the signal amplitude and frequency of intracellular Ca(2+) oscillations and intercellular Ca(2) wave elicited by specific acupuncture may contain information of the meridian routes and physiological function regulations. Through its binding proteins, Ca(2+) participates in almost all the physiological processes such as reproduction, metabolism, muscle contraction to cognition and memory. It is therefore hypothetically believed that Ca(2+) and inositol trisphosphate (IP(3), which is required for the propagation of intercellular Ca(2+) waves) can be the long-quested information carriers of the meridian physiological network, and they in conjunction with the gap-junction protein (connexon) build the material bases of the meridian. The author also specifies some possible directions of further study and experimental methods. PMID- 12754115 TI - [Ultrastructural observation of rat thymus tissue with coronavirus infection]. AB - OBJECTIVE: To investigate the ultrastructure of rat thymus tissues with rat coronavirus (RCV) infection for clarifying the mechanism responsible for the morphological changes of the cells infected by RCV. METHODS: Routine electron microscopy was performed for observing RCV-infected rat thymus tissues. RESULTS: Following RCV infection, endoplasmic reticulum (ER) pools of different dimensions were observed in the cytoplasm of the thymic epithelial reticular cells, merging subsequently with each other into larger ER lakes filled with particles of mature RCV, or viral inclusion bodies. After germination on the ER membrane, the viruses entered the matrix of the ER lake to mature and were eventually excreted to the extracellular space. The RCV particles were spherical in shape with a diameter of 100-130 nm and two distinct membranes, the outer one being the envelope and the inner one the nuclear capsid to enclose the viroplasm. Between the envelop and nuclear capsid was a electron-lucent middle layer comprising one to two thin membranous structures. Large quantity of short spike-like projections starting from the nucleus capsid penetrated the middle layer and the envelop to reach the glycoprotein coat and formed a corona-like structure. Mature RCV particles were distributed around the ER pools, cytoplasm, and intercellular space, and the RCVs in the endosome/lysosome were devoid of the envelop and nuclear capsid. CONCLUSION: The ER lakes are involved in the maturation of the viruses, and the envelop and nuclear capsid of the virus entering the cells from extracellular space are removed and degraded in the endosome/lysosome. Replications of virus occurs in plasma of the thymic epithelial reticular cells, and no RCV can be detected in the thymocytes. PMID- 12754116 TI - [In vitro degradation and subsequent biomechanical changes of poly(lactide-co glycolide) scaffolds prepared by mild heating under high pressure]. AB - OBJECTIVE: To study the changes in biomechanics and such indices as intrinsic viscosity poly (lactide-co-glycolide) (PLGA) scaffolds produced by mild heating under high pressure after in vitro degradation. METHODS: PLGA scaffolds with the porosity of 90.0% and 92.5% respectively were immerged in 37 degrees Celsius; saline for 8 weeks, and the changes in their mass, intrinsic viscosity and loss of compressive strength were assessed on a weekly basis, and the acidity of the degradation solution was also measured regularly. RESULTS: Significant differences was noted in the mass reduction between the scaffolds, and the intrinsic viscosity began to decrease in both groups in the first week to half of the original value till the sixth week. A 50% reduction in the compressive strength of the scaffolds occurred at the fourth week, and till the eighth week, obvious structural collapse was observed. Along with the changes, the acidity of the degradation solution increased from 6.0 to 6.5, and the solution of 90.0% porosity group had lower pH value during the first 4 weeks than 92.5% porosity group, but such difference was no longer seen afterwards. CONCLUSIONS: PLGA scaffolds made by mild heating under high pressure have stable biomechanical performance with the half-life of approximately 6 weeks, which can be applicable for tissue engineering. PMID- 12754117 TI - [Gene sequence analysis of SARS-associated coronavirus by nested RT-PCR]. AB - OBJECTIVE: To explore an effective means for the detection of Severe Acute Respiratory Syndrome (SARS)- associated coronavirus. METHODS: The RNAs of the virus contained in the sputum samples from established SARS patients were extracted and reversely transcripted, followed by nested PCR using the reversely transcripted cDNA as the template. The PCR products were cloned then into the pMD18-T vectors, followed by sequence analysis. RESULTS: Specific fragments were amplified from the sputum samples of SARS patients, which were confirmed by DNA cloning and sequencing to belong to SARS-associated coronavirus. The Result of Blast shows only the difference in one nucleic acid from the TOR2 strain of SARS associated coronavirus. CONCLUSION: Sequence analysis has confirmed the existence of SARS-associated coronavirus in the sputum samples of SARS patients, and nested RT-PCR is a quick, easy, and convenient way for the detection of the virus. PMID- 12754118 TI - [Clinical detection of polymerase gene of SARS-associated coronavirus]. AB - OBJECTIVE: To analyze the heterogeneity of polymerase gene fragment of SARS associated coronavirus from SARS patients, and establish a RT-PCR method for detecting SARS-associated coronavirus. METHODS: RT-PCR was performed using SARS coronavirus-specific primers to amplify the polymerase gene fragment of SARS associated coronavirus from specimens of suspected and established SARS cases. The amplicons were cloned and sequenced. All the obtained sequences were compared with the sequence of published SARS-associated coronavirus, and alignment was proceeded with other coronavirus sequences. RESULTS: Specific amplicons can be amplified from the sputum samples, throat swab and plasma of most SARS patients, and 8 were random selected and sequenced. All of them possessed 100% homology with the published SARS-associated coronavirus sequence, while all the negative controls were RT-PCR negative. Nucleotide-sequence and amino acid-sequence alignment of the fragment BNI109 with other six known coronavirus show that the fragment BNI109 is more close to bovine coronavirus(BCV) and murine hepatitis virus(MHV). The BNI109 fragment showed 75% homology with BCV and MHV at amino acid level. CONCLUSION: The polymerase fragment BNI109 of SARS coronavirus is highly conservative and is suitable for detecting SARS-associated coronavirus using RT-PCR method. PMID- 12754119 TI - [Isolation of rat tracheal smooth muscle cells and electrophysiological examination of the L- type calcium channel]. AB - OBJECTIVE: To establish a method for acute enzymatic isolation of rat tracheal smooth muscle cells (TSMCs) and study the electrophysiological properties of the L-type calcium channel of the cells. METHODS: Single rat TSMC was isolated by means of pronase E, followed by recording the electric currents in the single calcium channel using patch-clamp technique with cell attached configuration. RESULTS: Large amount of viable TSMCs were isolated through this method, characterized by normal cell morphology and easy detection of the L-type calcium channel activity in the cells. CONCLUSION: This method is relatively simple for high-yield isolation of TSMCs with normal morphology. PMID- 12754120 TI - [Clinical analysis of 15 cases of pulmonary sarcoidosis]. AB - OBJECTIVE: To facilitate the diagnosis and treatment of pulmonary sarcoidosis. METHOD: The clinical data of 15 cases of pulmonary sarcoidosis were retrospectively analyzed. RESULTS: Among these patients, 7 were identified with respiratory symptoms as the initial clinical representations, and 6 patients initially presented extrapulmonary symptoms, with 2 patients having no discomfort. Chest X-ray and lung CT scan results of 13 cases showed symmetrical lymph node enlargement in the bilateral lung hilus and mediastinum. Twelve patients had increased serum angiotensin-converting enzyme levels, and 10 had increased erythrocyte sedimentation rate, with only 1 case showing positive result in purified protein derivative skin test. CONCLUSION: Pulmonary sarcoidosis has atypical clinical symptoms, and the diagnosis should be reached by a careful evaluation of the clinical representations in conjunction with X ray, CT, and laboratory test results. PMID- 12754121 TI - [Absorption and distribution of 5-aminosalicylic acid from its chitosan capsule degraded by colon bacteria-released enzymes in rats]. AB - OBJECTIVE: To examine the absorption and distribution of 5-aminosalicylic acid (5 ASA) in rat colon after oral administration of its chitosan capsule. METHODS: 5 ASA in chitosan capsules (4.8 mg per 3 capsules) were administered orally in rats via a polyethylene cannula under light ether anesthesia. The rats in control group were given 1 ml 5-ASA carboxymethyl cellulose suspension (4.8 mg). Blood samples were obtained from the rat hearts and the colon tissues sampled at a given interval to measure the concentration of 5-ASA by HPLC. RESULTS: The area under the curve (AUC(0-12)) in the serum samples of chitosan capsule group was 0.62 times that of the control group, while in the colon tissue, the AUC(0-12) of chitosan capsule group was as much as 3.62 times that of the control group. CONCLUSION: The chitosan capsule may be a useful carrier of 5-ASA for colon specific delivery. PMID- 12754122 TI - [High-dose glucose induces human retinal endothelial cell apoptosis]. AB - OBJECTIVE: To examine the direct effect of high glucose levels on primary cultured human retinal capillary endothelial cells (HRCEC). METHODS: HRCECs were isolated from donated eyes and cultured for 6 days in the media containing 5 or 25 mmol/L glucose. The cell viability was determined by trypan blue exclusion assay and cell cycle analyzed by flow cytometry, with the cell apoptosis assayed by TUNEL method. RESULTS: The cell viability was significantly decreased after exposure to 25 mmol/L glucose, and the number of apoptotic cells determined by flow cytometry and TUNEL was significantly increased in response to high-dose glucose treatment. CONCLUSION: High-dose glucose induces apoptosis in HRCEC, which may contribute to the development of diabetic retinopathy. PMID- 12754123 TI - [Intravenous contrast imaging in diagnosis of acute renal allograft rejection: a comparative study with color flow Doppler imaging]. AB - OBJECTIVE: To assess the value of intermittent contrast second harmonic imaging (SHI) incorporating acoustic densitometry in the diagnosis of acute renal allograft rejection in comparison with color Doppler flow imaging (CDFI) for determining the resistance index (RI). METHODS: Eight canine models of acute renal allograft rejection were established, subjected subsequently to examinations with SHI and acoustic densitometry to determine the acoustic density. Color flow Doppler was also performed to determine RI, and serum creatinine (Cr) levels were measured. RESULTS: The time-intensity curve (TIC) showed that the area under the curve, the peak intensity (PI) and RI all had linear correlation with serum Cr levels, with the correlation coefficient gamma of 0.978, 0.972 and 0.708 respectively (by SPSS10.0). CONCLUSION: Intermittent contrast SHI is effective to evaluate the perfusion of renal allograft, and when combined with acoustic densitometry, the resultant TIC parameters are in closer correlation with acute renal allograft rejection than RI. PMID- 12754124 TI - [Mechanical strength and in vivo degradation of human hair keratin-polylactic acid composite rods]. AB - OBJECTIVE: To test the mechanical strength and observe the in vivo degradation of self-made human hair keratin-polylactic acid (HHK-PLA) composite rods designed for use in internal fixation. METHODS: Twenty such HHK-PLA composite rods were tested for shear strength, bending strength and bending modulus using the material testing system MTS-858 Mini Bionix. A total of 36 samples of HHK-PLA composite rods designed for internal fixation of bone fracture were randomly implanted in dorsal subcutaneous tissue of 18 SD rats, and weight losses of these rods were measured 1, 2, 4, 8, 12, 16, 20, 24 and 28 weeks after the implantation to evaluate the in vivo degradation of the material. RESULTS: The initial shear strength of HHK-PLA rod was 241 MPa, bending strength 358 MPa, and bending modulus 13 GPa. The test demonstrated a slower rate of degradation in SD rats in earlier period following implantation than in later period. CONCLUSION: HHK-PLA composite rods have good initial mechanical strength and tolerable degradation in vivo, and may be used potentially for internal fixation of the weigh-bearing bones of the limbs. PMID- 12754125 TI - [Calcification and biomechanics in vivo of ultra-microporous expanded polytetrafluoroethylene in rabbits]. AB - OBJECTIVE: To study in vivo calcification and biomechanics of the ultra microporous expanded polytetrafluo- roethylene (UePTFE) in rabbits in comparison with glutaral-treated bovine pericardium (BP), so as to assess the potential of UePTFE as a material for cardiac valve prosthesis. METHODS: Factorial analysis was adopted in the experiment. UePTFE and glutaral-treated BP of appropriate sizes were embedded beneath skin of young New Zealand rabbits, and at 0, 1, 3 and 6 months following the implantation, the materials were measured for the content of calcium and biomechanics properties. RESULTS: Lower level of calcification of the UePTFE occurred after the implantation, as compared with BP, and the biomechanics indices of the former UePTFE were obviously suprior to those of the latter. CONCLUSION: UePTFE is a better material than BP for cardiac valve prosthesis. PMID- 12754126 TI - [Apoptosis-inducing effect of palmitic acids on rat pancreatic islet cells in primary culture: a preliminary study]. AB - OBJECTIVE: To investigate the in vitro apoptosis-inducing effect of palmitic acid on pancreatic islet cells in primary culture, thereby to understand the role of palmitic acid in the pathogenesis of type 2 diabetes mellitus. METHOD: SD rat pancreatic islet cells were isolated and cultured in monolayer in vitro followed by incubation with stepwise diluted palmitic acid (0, 0.125, 0.25 mmol/L and 0.5 mmol/L respectively), and the insulin concentrations in the culture medium were determined by radio immunological methods. Morphological observation with a fluorescence microscope was conducted after double staining of the cells with PI/Hoechst 33342. RESULTS: The glucose-stimulated insulin secretion was inhibited by palmitic acid at the concentration of 0.25 and 0.5 mmol/L, which also induced obvious cell apoptosis. CONCLUSION: Palmitic acid is capable of inducing islet cell apoptosis in a dose-dependent manner. PMID- 12754127 TI - [Pathological changes in rabbit brain after traumatic head injuries with seawater immersion]. AB - OBJECTIVE: To observe the pathological impact of seawater on rabbit brain tissue with severe traumatic injury. METHODS: Modified rabbit models of severe brain injury was utilized, in which the damaged brain tissues were subjected to immersion with fresh seawater for 30 min. A control group was set up in which the trauma was induced without subsequent seawater immersion. Tissue sampling was performed at the brain injury sites at 0, 3, 8 h after seawater immersion and the pathological changes in the brain tissues were observed by means of HE staining. RESULTS: Severe traumatic brain edema occurred in both of the two groups, but the onset time of edema differed. In the control group, brain edema was obvious at 3 h after treatment and hardly aggravated at hour 8; while in seawater treatment group, severe brain edema occurred at 8 h after the treatment and aggravated progressively. CONCLUSION: Seawater immersion delays the onset and peak of traumatic brain edema following severe brain injuries, but can eventually aggravate the traumatic edema. PMID- 12754128 TI - [Scanning electron microscope observation of the effect of different dentin bonding surfaces on the bonding interface]. AB - To study the microstructural influence of dry or wet dentin bonding surface on the dentin bonding interface, five wet bonding systems were used to bond Chrisma B20 resin with dry or moist dentin surface with scanning electron microscope. All these systems were found to be well able to infiltrate into the dentin bonding interface on wet dentin surface, and significant changes occurred in the bonding interface when acetone-based adhesives were applied on dry dentin surfaces, demonstrating thinner hybrid layer and the formation of partially hybrid region, while alcohol- and water-based adhesives showed no changes. The necessity of moist dentin surface is therefore suggested when wet bonding systems are used. PMID- 12754129 TI - [Association between glutathione S-transferase M1 gene deletion and genetic susceptibility to endometriosis]. AB - OBJECTIVE: To evaluate the possible association of the glutathione S-transferase M1 (GSTM1) gene polymorphism with the susceptibility to endometriosis in women of Han nationality in Guangdong Province. METHODS: Polymerase chain reaction was used to identify the GSTM1 genotypes in 76 patients with endometriosis and 80 controls (surgical patients for gynecological problems other than endometriosis). RESULTS: The frequencies of the GSTM1 null genotypes in patients with endometriosis and controls were 65.8% and 46.3%, respectively, showing a significant difference between the endometriotic cohort and the control group (X(2)=6.03, P < 0.05). Individuals with GSTM1 null genotype were exposed to risks for endometriosis 2.24 times that of subjects without these genotypes OR=2.24, 95% CI=1.17-4.27 . CONCLUSION: GSTM1 gene deletion might bea risk factor for endometriosis in women of Han nationality who are native residents in Guangdong Province. PMID- 12754130 TI - [Methotrexate in conjunction with meloxicam for treating ankylosing spondylitis: a clinical observation of the efficacy and safety]. AB - OBJECTIVE: To evaluate the efficacy and safety of combined use of methotrexate and meloxicam for treating of ankylosing spondylitis (AS), and to determine the optimal dosage of methotrexate. METHODS: Thirty-two patients with AS were divided into group A (n=14) to receive oral methotrexate at the dose of 12.3+/-3.6 mg once a week and group B (n=18) at the dose of 21.3+/-3.2 mg once a week. Each of the patients in both groups also took oral meloxicam (7.5 mg) for 3 months. RESULTS: After treatment, the clinical indexes and inflammatory parameters of the patients significantly improved in both groups, with a total efficacy of 78.6% in group A and 88.9% in group B. CONCLUSION: The combination of methotrexate with meloxicam may achieve satisfactory results in AS treatment, which is not positively related to methotrexate dosage. In short-term usage, meloxicam does not increase the toxicity of methotrexate. PMID- 12754131 TI - [Transforming growth factor-beta1, estradiol, progesterone and lutropin levels in follicular fluid after ovarian stimulation]. AB - OBJECTIVE: To investigate the role of transforming growth factor-beta1 (TGFbeta1) and the sex hormones in the follicular fluid (FF) on the day of ovum pick-up (OPU) during controlled ovarian hyperstimulation (COH) cycles. METHODS: FF and the oocytes were obtained from the follicles of 90 women undergoing ovulation stimulation in vitro fertilization (IVF) treatment. TGFbeta1, estradiol, progesterone, and lutropin concentrations in the FF samples collected during transvaginal oocyte retrieval were measured using enzyme-linked immunosorbent assay (ELISA). The maturity and fertilization of the oocytes were observed, and ultrasonography performed to confirm clinical pregnancies 4 weeks after the embryo transfer. RESULTS: In the FF containing mature oocytes, progesterone and lutropin concentrations were significantly higher than those in the FF with immature oocytes. The mean concentrations of TGF beta1, progesterone and lutropin in the FF from fertilized subjects were obviously higher than those in the FF from non-fertilized subjects (P < 0.05), and in subjects with pregnancy, higher mean concentrations of TGFbeta1 and lutropin were detected as compared with the concentrations measured from non-pregnancy subjects (3 631.4+/-1 426.3 pg/ml and 0.74+/-0.25 mIU/ml vs 2 189.2+/-1 180.4 pg/ml and 0.52+/-0.29 mIU/ml respectively, P < 0.05). Estradiol concentrations in the FF seemed to undergo no obvious changes during the whole procedure, and evinced no significant differences between the groups. CONCLUSION: Higher lutropin and progesterone concentrations in the FF on the day of OPU may promote oocyte maturation, while TGFbeta1 and lutropin levels appear to be associated with the maturation and fertilization of the oocytes, and may be indicative of the IVF outcome. PMID- 12754133 TI - [Assay of Acinetobacter SPP drug-resistance by Kirby-Bauer and Etest method]. AB - OBJECTIVE: To study the drug-resistance of Acinetobacter spp and observe whether antibiotic-beta- lactamase inhibitor complexing agent enhances the sensitivity of the drug-resistant bacteria to the antibiotics. METHODS: Susceptibility tests to the antibiotics were performed for 60 isolated strains of Acinetobacter spp with Kirby-Bauer (K-B) and Etest methods. RESULTS: The drug-resistance rate of the strains to cefoperazone, ampicillin, ticarcillin and piperacillin exceeded 55.0%, and antibiotic-beta-lactamase inhibitor complexing agents evinced better antibacterial activity than antibiotics used alone (P < 0.001). The 2 antibiotic complexes incorporating sulbacta had more potent antibacterial activity than the complexes incorporating clavulanate (2 agents) and tazobactam (1 agent), with the mean ranks of 7.38, 6.43, 5.57, 5.47 and 5.50 respectively. Cefoperazone combined with sulbacta produced a reduction in MIC(90) and MIC(50) that were only 4.69% and 0.59% of the MIC(50) of cefoperazone, and in addition, sulbacta caused cefoperazone- and ampicillin-resistant rates to reduce from both 75.0% to 5.0% and 24.0%, respectively. CONCLUSION: Acinetobacter spp is resistant to a wide spectrum of commonly-used antibiotics, but cefoperazone in combination with sulbacta can obviously enhance antibacterial potency, suggesting the primary role of this regimen in fighting Acinetobacter spp infections in hospital. PMID- 12754132 TI - [Relationship of serum iron and ferritin with the indicators for hepatic fibrosis]. AB - OBJECTIVE: To study the relationship of serum iron and ferritin with the indicators for hepatic fibrosis and hepatic iron overload. METHODS: Liver tissue specimens were obtained from 41 patients with benign (16) or malignant (25) liver diseases by 1 second liver biopsy, and routine microscopic examination was performed after haematoxylin-eosin (HE) and Perl's Prussian staining. Atomic absorption spectrum, radioimmunoassay and enzyme-linked immunosorbent assay were respectively employed to examine the serum levels of iron, ferritin, hyaluronic acid, laminin, human procollagen type , and collagen type . RESULTS: Between patients with benign and malignant liver diseases, significant differences were found in the serum ferritin levels (P < 0.05), but not in serum iron levels (P > 0.05). It was also noted that the levels of the indicators for hepatic fibrosis in patients with benign and early-stage malignant diseases varied significantly from the levels in normal subjects, but these differences were not observed between normal subjects and patients with end-stage hepatic malignancies. Serum iron and ferritin were found to be associated with serum laminin levels (serum iron: r=0.439, P=0.031; serum ferritin: r=0.476, P=0.016), and no iron granules detected in the tissue specimens of hepatocellular carcinoma. CONCLUSIONS: Most of the patients with hepatocellular carcinoma have elevated serum ferritin levels. The serum levels of iron and ferritin are statistically correlated with serum laminin level. Obvious reduction of iron content is typical of hepatic malignant tissues in comparison with the benign tissues, and the reduction in the levels of the indicators for hepatic fibrosis might involve the inhibition of collagen synthesis in the tumor tissues from patients with end-stage hepatocellular carcinoma. Most of the cases of alcoholic fatty liver are complicated by liver iron overload, often marked by serum iron and ferritin levels. PMID- 12754134 TI - [Analysis of the relation between serum immunoglobulin and auto-antibody levels in patients with rheumatoid arthritis]. AB - OBJECTIVE: To explore the relationship between serum immunoglobulin (Ig) levels and the positivity rate for auto- antibodies in patients with rheumatoid arthritis (RA). METHODS: The levels of immunoglobulins (IgG, IgM, IgA) and rheumatoid factor (RF) were measured using Olympus AU600 system, and indirect immunofluorescence and immunoblotting methods were employed respectively for detecting the autoantibodies in the serum from 40 RA patients and 30 normal adults. Statistical analysis of the relation between the Ig levels and autoantibodies was performed. RESULTS: Compared with normal control group, significantly higher serum IgA, IgG and IgM levels were detected in RA patients (P < 0.01), and at least one autoantibody was present in the serum sample from 26 out of the 40 RA patients, who had the highest RF positivity rate of 20% (8 cases). The serum IgA level in autoantibody-positive patients was significantly higher than that in autoantibody-negative ones (P < 0.01), and in the 18 patients with serum IgA level beyond the normal range, 15 (83.3%) were positive for autoantibody, while in the 22 patients with normal serum IgA level, the autoantibody positivity rate was only 50% (11/22), showing significant difference (P < 0.05). CONCLUSION: Serum immunoglobulins A (IgA) level is closely associated with the positivity rate for autoantibodies in RA patients, and RA patients with serum IgA level beyond the normal range was more likely to be positive for the autoantibodies. PMID- 12754135 TI - [Effects of dental cements on the stress distribution in amalgam-restored teeth]. AB - OBJECTIVE: To evaluate the effects of 5 kinds of base materials on the stress distribution and deflection in amalgam- restored teeth. METHODS: A computerized two-dimensional finite-element of the tooth model first molar was established. Various dental cements with a thickness of 0.5 mm and amalgam restorations with thickness of 2 mm in the prepared cavity were simulated and subjected to microhardness test (load=45 kg). The stresses and deflections along the cavity wall interface were analyzed by finite-element method. RESULTS: High stress and deflection occurred mainly at the line angle of the cavity, this could be magnified by using cements with lower elasticity modulus. The maximum compressive stress at the line angle induced by common ZOE or Ca(HO)(2) was increased 3.3 times or 2 times and deflections increased by 40% and 25% respectively that by Zinc phosphate and glass ionomer. CONCLUSION: Modulus elasticity of cement plays an important role in affecting the fracture strength of the amalgam restored tooth. The lower the elasticity modulus, the higher the stresses and deflections induced in the dentine. From the point of mechanical property, zinc phosphate is one of the best dental cements. PMID- 12754136 TI - [Expressions of angiotensin II type 1 receptor mRNA in fibrotic and normal liver tissues]. AB - OBJECTIVE: To investigate the expression profile of angiotensin type 1 receptor (AT1R) mRNA in fibrotic and normal liver tissues. METHODS: Semi quantitative RT-PCR was used to detect the expression of AT1R mRNA in 18 specimens of fibrotic liver tissues adjacent to liver cancers and 12 normal liver tissue specimens. Specific AT1R product and GAPDH product used as internal control were both amplified by RT-PCR, and the ratio of their integrated optical densities calculated to estimate the relative quantity of AT1R mRNA expression. RESULTS: The relative mRNA expression of AT1R in fibrotic liver tissues was significantly higher than that of normal liver tissues (1.27+/-0.45 vs 0.71+/ 0.21, P < 0.01). CONCLUSION: AT1R may play an important role in the development of hepatic fibrosis. PMID- 12754137 TI - [CT findings and risk factor analysis of hepatic injury induced by three dimensional conformal radiation therapy]. AB - OBJECTIVE: To investigate the CT findings and define the risk factors for early hepatic injury induced by three- dimensional conformal radiation therapy in patients with primary liver cancer. METHOD: This study recruited 52 patients with primary liver cancer undergoing photon beam radiation at a total dose of 35 to 65 Gy (49.88+/-7.23 Gy) completed within 18 to 31 d. CT examinations were performed within 1 to 3 months after the completion of the therapy, and logistic regression was used to analyze the CT features of early hepatic injury, in an attempt to define the correlation of the injury occurrence with such factors as the patients' age, gender, treatment portal numbers, liver cirrhosis, transcatheter arterial embolization (TACE), postoperative recurrence, total radiation dose, target volume and fractional dose. RESULTS: Thirty-one (59.6%) patients showed CT features of hepatic injury, displayed as the area with hypodensity in consistency with the radiation coverage. The risk factors correlated to the injury occurrence included the total dose, target volume and fractional dose, but not the patients'age, gender, treatment portal number, liver cirrhosis, TACE, postoperative recurrence differences and radiation-induced hepatic injury. CONCLUSION: Early radiation-induced hepatic injury is related to fractional dose, total dose and target volume adopted in radiation therapy. Higher fractional dose, total irradiated dose and larger target volume may result in increased risk of injury. PMID- 12754139 TI - [An epidemiological study of 2,353 patients injured in motor vehicle accidents]. AB - OBJECTIVE: To investigate the clinical conditions of the in-patients with motor vehicle injury and survey the distribution features of the injuries. METHODS: The clinical records of 2,353 patients with motor-vehicle injuries who were hospitalized in a hospital in Shaoyang City of Hunan Province, China, within the period from 1997 to 2001, were collected and surveyed. The patients were diagnosed according to International Classification of Diseases-9th Revision (ICD 9) codes and statistical analysis was performed in an attempt to define the injury features of this cohort of patients. RESULTS: The patients with motor vehicle injuries took up a fraction 3.16% of the total in-patients treated in this hospital (74,368) within the defined period, with a male to female ratio of 2.37:1.00. Subjects in the age range of 15 to 44 years were most likely to be involved in the accidents (55.9%), and in terms of occupation, farmers who lived outside the urban regions had the highest ratio in the total injured patients (34.0%). In the motor-vehicle accidents, the major injuries occurred in the head, usually with moderate severity when admitted into hospital (69.9%). CONCLUSION: Traffic accident is a crucial public health issue that severely affects the life quality of the wounded. Effective legislation and regulations should therefore be instituted, and relevant health education program be launched to control and prevent the occurrence of these accidents. PMID- 12754138 TI - [Significances of plasma levels of neuropeptide Y pre- and post-treatment in patients with decompensated cirrhosis and ascites]. AB - OBJECTIVE: To observe changes of plasma neuropeptide Y(NPY) level in ascitic patients with decompensated cirrhosis pre- and post-treatment, there fore to understand the correlation of NPY with liver function and ascitic formation. METHODS: NPY plasma levels in 20 patients with decompensated cirrhosis and ascites were detected by radioimmunological assay. RESULTS: Plasma NPY levels in patients with cirrhotic ascites were significantly lower than those in normal subjects(P < 0.01) in spite of the increases of NPY after ascites decreased (P < 0.01), which were still lower than those in normal subjects(P < 0.01). CONCLUSION: Decreased plasma NPY levels are correlated with the severity of liver damage and may be responsible for the changes of hemodynamics and ascitic formation in patients with liver cirrhosis. PMID- 12754140 TI - [Fresh amniotic membrane transplantation for ocular surface diseases]. AB - OBJECTIVE: To evaluate the therapeutic effects of fresh amniotic membrane transplantation for ocular surface disorders (OSDs). METHODS: Fresh amniotic membrane transplantation was performed in 38 cases (52 eyes) of ocular surface diseases, including pterygium, recurrent pterygium, Mooren's ulcer, Viral keratitis, and symblepharon. Follow-up studies ranging from 3 to 12 months were conducted. RESULTS: No acute graft rejection was observed following the operation in these cases, and no recurrence was found in cases of pterygium and corneal ulcer during the follow-up period. The ocular movement was restored for the eyes with severe symblepharon. CONCLUSION: Fresh amniotic membrane can be used as a graft material for ocular surface reconstruction, in which complete removal of the pathological tissues and reliable fixation of the amniotic membrane graft are crucial steps. PMID- 12754141 TI - [Sclerotherapy for renal cyst using erythromycin and procaine via renal puncture guided by B-ultrasound: report of 1,000 cases]. AB - Our experience of sclerotherapy for renal cyst injection of erythromycin and procaine via ultrasound-guided renal puncture is reported. The total efficiency reached 96% with low recurrence rate in the 1,000 cases receiving this therapy, suggesting that this approach is a simple, accurate and effective renal cysts with minimized postoperative complications treatment. PMID- 12754142 TI - [Preparation of egg yolk immunoglobulin (IgY) against p53 protein expressed in E.coli]. AB - OBJECTIVE: To isolate immunoglobulin (IgY) from egg yolk of p53 protein-immunized hens and to study its reactivity to the antigens. METHOD: Immunization of egg laying hens was performed for 3 times at the interval of 14 days with purified p53 protein that had been expressed in E.coli. The eggs laid by these immunized hens were then collected during the whole period of the experiment to prepare IgY from egg yolk by ammonium sulfate precipitation. Final purification and identification of the IgY were performed using SDS-PAGE, enzyme-linked immunosorbent assay and Western blotting. RESULT: All immunized hens developed specific antibodies to p53 protein in contrast to the control ones. The highest titers of the IgY occurred 4 weeks after the first immunization and reached 1:10(6), which remained stable for up to 3 months. CONCLUSION: IgY technology is less costly, non-invasive, fast, simple and highly efficient to generate polyclonal antibodies. PMID- 12754143 TI - [Prophylaxis of hepatic veno-occlusive disease by low-molecular-weight heparin and lipo- prostaglandin E1 after allogeneic peripheral blood stem cell transplantation]. AB - To study the effects of low-molecular-weight heparin and lipid microspheres containing prostaglandin E(1) (lipo PGE) in preventing hepatic veno-occlusive disease after allogeneic peripheral blood stem cell transplantation, we observed 21 cases of allogeneic stem cell transplantation, in which the combined treatments were administered from day -7 to day +30. As a result, only 2 of these cases developed hepatic veno-occlusive disease, without obvious treatment-related adverse effects, suggesting the safety and effectiveness of the combined treatment using low-molecular-weight heparin and lipo PGE1 in the prevention of hepatic veno-occlusive disease after allogenetic stem cell transplantation. PMID- 12754144 TI - [Expression of matrix metalloproteinase-2 in hypopharyngeal carcinoma and its clinical significance]. AB - OBJECTIVE: To study the relationship between matrix metalloproteinase-2(MMP-2) and the biological behavior, metastasis and prognosis of hypopharyngeal carcinoma. METHODS: Two-step immunohistochemical method was used to examine the expression of MMP-2 mAb in 32 specimens of hypopharyngeal carcinoma tissues and adjacent normal mucosa. Statistical comparison was performed after grouping. RESULTS: It was found that the positivity rate for MMP-2 in the hypopharyngeal carcinoma tissues was 53.1%(17/32). Significant differences were noted when the expression levels of the mAb were compared between the normal mucosa adjacent to the carcinomas and the carcinomatous tissue, and between non-metastatic carcinomas and those with cervical lymph node metastasis. Patients with a positive result had obviously lower 3-year survival rate than those with negative results. CONCLUSION: MMP-2 may play a role in the lymph node metastasis of hypopharyngeal carcinoma, and is likely to be indicative of the patients' prognosis. PMID- 12754146 TI - [Construction of recombinant adenovirus using modified AdEasy system]. AB - To simplify AdEasy system for better efficiency, we attempted some modifications on the original system. Specifically, the Pme -linearized plasmid pAdtrack was transformed into competent AdEasier-1 cells prepared from treatment with CaCl(2). The DNA contained in the identified recombinant plasmid was digested with Pac and transfected into 293 cells to package the adenovirus, followed by identification of the recombinant adenovirus by means of observation of green fluorescence protein expression under fluorescence microscope. Chemical transformation of the linearized plasmid into AdEasier-1 cells resulted in very high success rate (20/20) for producing positive recombinant adenovirus clones, confirming the efficiency of the modified AdEasy system in constructing recombinant adenovirus. PMID- 12754145 TI - [Detection of nuclear factor kappa B by immunochemical staining and image analysis]. AB - OBJECTIVE: To find a relatively simple and safe method for quantitative assay of nuclear factor (NF)-kappa B activity. METHODS: Immunochemical staining of NF kappa B/p65 subunit with its antibody was performed in human umbilical vein endothelial cells stimulated with advanced glycation end products-modified human serum albumin (AGE-HSA). The ratio of p65 subunit staining in the nuclei and cytoplasm was determined by means of imaging analysis, and the results were compared with those of electrophoretic mobility shift assay (EMSA). RESULTS: NF kappa B could be activated by AGE-HSA in a time- and dose-dependent manner, and the results obtained from immunochemical staining were consistent with those from EMSA. CONCLUSION: p65 immunochemical staining and subsequent image analysis is feasible in the quantitative detection of NF-kappa B activation. PMID- 12754147 TI - [Application of self-controlled breathing technique in respiratory gating that triggers the shockwave for lithotripsy of urinary calculi]. AB - OBJECTIVE: To review our experience with the application of respiratory gating technique in extracorporeal shock wave lithotripsy (ESWL). METHODS: According to the established criteria 424 patients with urinary calculi were included in this study to received ESWL. These patients were divided into three groups, in which different breathing techniques were used for ESWL: group A with free breathing, group B with basically normal breathing except for the breath-holding upon each shockwave triggering, and group C with controlled breathing acquired from preoperative training. All the patients were followed up from 3 to 6 months, during the period of which the effect of ESWL was observed and recorded for comparative appraisal of the therapeutic efficacy. RESULTS: The therapeutic effect significantly differed between the groups (P < 0.01), with the best effect recorded in group C (96.40%), followed by group A (83.87%) immediately before group B (86.67%), indicating that the essential difference occurred between group C and groups A and B, but not between the latter two groups. None of the patients developed serious short-term complications. CONCLUSION: The application of breathing technique in respiratory gating for ESWL is safe and effective without causing serious complications. PMID- 12754149 TI - [Therapeutical effect of Zhiranxiao tablets on internal hemorrhoid hemorrhage: a clinical observation in 569 cases]. AB - OBJECTIVE: To investigate the therapeutic effect of Zhiranxiao tablets derived from compound Zhiranxiao decoction, a preparation from traditional Chinese medicine, on internal hemorrhoid hemorrhage. METHODS: The treatment group consisted of 596 patients with internal hemorrhoid hemorrhage who received oral Zhiranxiao tablets in a course of 6 d for treatment, and another 60 such patients were given a different preparation from traditional Chinese medicine, Zhining tablets, to serve as the control group. The volume of hemorrhage was recorded and scored before and after the treatment to evaluate the therapeutic effect of the tested dosage forms. RESULTS: The scores for hemorrhage in both groups were significantly decreased after the administration of the two dosage forms, but no statistical difference was observed (P > 0.05) between the two groups. CONCLUSION: Zhiranxiao tablets are also effective in the treatment of internal hemorrhoid hemorrhage. PMID- 12754148 TI - [Acute respiratory tract obstruction during thyroid operation: analysis of 10 cases]. AB - OBJECTIVE: To explore the causes and management of acute respiratory tract obstruction in patients undergoing thyroid operation. METHODS: A retrospective review was conducted in 10 such cases that we encountered in our 11 years' experience with thyroid operation. RESULTS: Respiratory tract obstructions occurred in 10(0.31%) of 3,186 cases during thyroid operation. The causes for the obstructions included tracheomalacia in 2 cases, hematoma compression in 5 cases, trachea convulsion in 2 cases and phlegm obstruction in 1 case. Tracheotomy or intratracheal intubation was performed in 6 cases and hematoma removal in 4 cases, with the occurrence of death in 1 case. CONCLUSION: Hematoma compression and tracheomalacia etc. are among the most common causes of acute respiratory tract obstruction during thyroid operation, the occasion of which demands immediate implementation of tracheotomy or intratracheal intubation. PMID- 12754150 TI - [Causes of urinary tract infection and its prevention in patients with indwelling urinary catheter: report of 152 cases]. AB - OBJECTIVE: To investigate the causes of urinary tract infections occurring in patients with indwelling urinary catheter, and to find the preventive means against this condition. METHODS: The clinical data of 152 such cases admitted between March, 2000 and February, 2002 in our department were reviewed. RESULTS: In this group of patients, 57 (37.5%) developed urinary infections due to the indwelling catheter, a rate significantly lower than that among previous cases (46.6%, P < 0.05) treated in our department. CONCLUSION: Careful evaluation of the indications for indwelling urinary catheter, strict aseptic procedures and shortened indwelling time of the catheter may help prevent the occurrence of the infection. PMID- 12754151 TI - [Usage of articles in English medical papers]. AB - The authors attempt to grammatically classify the usage of articles, for the purpose of decreasing the misuse of articles, which frequently occurs in English medical paper written by Chinese authors. PMID- 12754152 TI - [Diagnostic value of transvaginal color Doppler sonography for endometrial cancer]. AB - OBJECTIVE: To study the values of transvaginal color Doppler sonography (TV-CDS) in the diagnosis of endometrial cancer. METHOD: In this study we selected 42 pathologically defined endometrial cancer cases, in which the preoperative TV- CDS results were reviewed and the myometrial invasion investigated in comparison with the postoperative histopathological findings, for extracting the ultrasonographic features of this malignancy in different clinical stages. RESULTS: TV-CDS yielded the diagnoses of endometrial cancer that were highly consistent (with a rate of 95.24%) with the pathological findings, also capable of detecting myometrial invasion in 93.33% of the cases. The depth of myometrial invasion is accurately defined by TV-CDS in 80.95% of the cases. CONCLUSION: TV CDS provides valuables evidence for the diagnosis and clinical staging of endometrial cancer. PMID- 12754153 TI - In memoriam Bela Bohus 1936-2000. PMID- 12754154 TI - Neurodevelopmental liabilities in schizophrenia and affective disorders. AB - There is now considerable evidence that both schizophrenia and affective disorders have their origin at least in part in events that occur during early pre- and post-natal development. In the case of schizophrenia, many observations, for example, increased risk for schizophrenia in the offspring of mothers who had influenza A during their second trimester of pregnancy and evidence for abnormal neuronal migration in the cerebral cortex of post mortem tissue from schizophrenic patients, suggest that a second trimester insult may have occurred and that this insult may have increased the risk for the development of schizophrenia in late adolescence or early adulthood. Animal studies have found that rats that undergo exocitotoxic damage to the ventral hippocampus on postnatal day 7 develop exaggerated sensitivity to dopamine-stimulating drugs or to stressful stimuli that becomes apparent after sexual maturity but not before, providing a neurodevelopmental model of schizophrenia. Similarly, post-weaning social isolation leads to enhanced responses to dopaminergic drugs and to stress that emerges after sexual maturity. These animal models are proving to be valuable tools to study the neurobiological mechanisms mediating the influence of early insults to the nervous system on later behavioural functions. In the case of affective disorders, although the evidence is not as strong, a number of the same observations have been made suggesting that an insult during early ontogeny may lead to the development of affective disorders later in life. For example, retrospective studies of people with affective disorders showed that they were more likely to have attained motor milestones at a later age and to have had poorer academic performance as children. There is a wealth of evidence suggesting hyperfunctioning of the hypothalamic-pituitary-adrenal (HPA) axis in affective disorders. Animal studies have shown that early maternal deprivation can lead to lasting changes in the reactivity of the HPA axis to stressful stimuli, providing another link from early experience to adult psychopathology. Continued studies of the effects of pre- and early post-natal events on the development of the nervous system and the relationships of these events to schizophrenia or affective disorder will provide new insights into the mechanisms underlying these common neuropsychiatric illnesses. PMID- 12754155 TI - Control of the serotonergic system by the medial prefrontal cortex: potential role in the etiology of PTSD and depressive disorders. AB - The prefrontal cortex is involved in an array of higher brain functions that are altered in psychiatric disorders. Serotonergic neurons of the midbrain rapbe nuclei innervate the prefrontal cortex and are the cellular target for drugs used to treat mood disorders such as the selective serotonin (5-HT) reuptake inhibitors. Anatomical evidence supports the existence of projections from the medial prefrontal cortex (mPFC) to the dorsal raphe nucleus (DR). We report on a functional control of the activity of DR 5-HT neurons by projection neurons in the mPFC. The stimulation of the mPFC elicits two types of responses in DR 5-HT neurons, orthodromic excitations and inhibitions. Excitations are mediated by AMPA/KA and NMDA receptors whereas inhibitions are mediated by GABA(A) and 5 HT(1A) receptors. The activation of a subgroup of 5-HT neurons increases 5-HT release which subsequently activates 5-HT(1A) autoreceptors on other 5-HT neurons. GABA(A)-mediated inhibitions involve GABAergic elements in the DR or adjacent areas. Pyramidal neurons of the mPFC co-express postsynaptic 5-HT(1A) (inhibitory) and 5-HT(2A) (excitatory) receptors. Consistent with the above observations, the selective activation of both receptors in mPFC reduced and increased, respectively, the firing activity of DR 5-HT neurons and the 5-HT release in mPFC. Overall, these data indicate that the activity of the 5-HT system is strongly controlled by the mPFC. Thus, the abnormal prefrontal function in post-traumatic stress disorder and depressive patients may induce a disregulation of 5-HT neurons projecting to other brain areas that can underlie the existing symptomatology in these psychiatric disorders. PMID- 12754156 TI - Risk factors for the neurohumoral alterations underlying personality disturbances. AB - Numerous studies have shown that MAO-B activity in platelets correlates with specific personality characteristics such as sensation seeking and impulsiveness. Low levels of platelet MAO as well as the personality traits associated with these low levels have been associated with type 2 alcoholism, recurrent criminality and antisocial violent behavior. Platelet MAO has a high degree of heritability and regulation of MAOB gene expression seems to explain most of the inter-individual differences in activity. The transcription factor family AP-2 is an important regulatory factor for neural gene expression and neural development, especially in midbrain structures, including the monoaminergic nuclei. In man, the gene encoding AP-2beta contains a polymorphic region in the second intron, consisting of a variable number of tandem repeats [CAAA](4-5). The long AP-2beta allele has previously been associated with specific personality traits as well as with binge-eating disorder characterized by an impulsive temperament. We have shown that males and females homozygous for the long AP-2beta allele display significantly lower platelet MAO activity compared to subjects with one or two short alleles. Thus, we find it likely that the personality disturbances previously linked to low platelet MAO activity could be associated with the presence of two long alleles of the AP-2beta gene. We suggest that the molecular mechanisms underlying the association between platelet MAO and vulnerability, e.g. substance abuse, may involve specific transcription factors that regulate the expression of midbrain monoamine structures as well as that of platelet MAO. PMID- 12754157 TI - Schizophrenia and bipolar disorder are distinguished mainly by differences in neurodevelopment. AB - This paper examines the commonalities and the differences between schizophrenia and bipolar disorder. Recent studies suggest a possible overlap in genetic susceptibility to the two conditions. However, while the influence of early environmental effects, particularly obstetric complications, has been established for schizophrenia, no such replicable association with bipolar disorder has been found. Structural abnormalities of the brain have been identified in both schizophrenia and bipolar disorder, but while the volume of the amygdala and hippocampus appears decreased in schizophrenia, this is not the case in bipolar disorder; indeed there are some suggestions of increased volume of the amygdala. Furthermore, schizophrenia is characterised by lower IQ, executive function and verbal memory, but there is little evidence of trait neuropsychological deficits in bipolar disorder. Similarly, premanic children do not show the cognitive and neuromotor impairments characteristic of those destined to develop schizophrenia. The most plausible explanation is that the two conditions share some genetic predisposition but differ in that schizophrenia but not bipolar disorder is subject to additional genes or early environmental hazards causing neurodevelopmental impairment. PMID- 12754158 TI - Multimodal neuroimaging studies and neurodevelopment and neurodegeneration hypotheses of schizophrenia. AB - The interpretation of the huge number of results in schizophrenia research using neuroimaging is uncertain. However, the simultaneous use of complimentary data obtained with these techniques may yield more relevant information in this regard. In this paper we present a series of studies performed by our group in two schizophrenic samples with the use of structural (magnetic resonance imaging, MRI), functional [glucose positron emission tomography (PET) and N-acetyl aspartate (NAA) magnetic resonance spectrocopy] and neurophysiological techniques (the P300 event-related potential). Transversal and longitudinal measurements were performed.The integrated vision of the results so obtained allows us to propose the hypothesis of a neurodevelopmentally determined state of prefrontal disinihibition, in which the degree of atrophy would directly relate to the metabolic rate. This state would already be present in the first stages of illness and could have neurotoxic consequences in the long term. This would explain the findings of an association between sulcal cerebrospinal fluid (CSF) and illness duration and decreased NAA levels in chronic but not in recent-onset cases. The prefrotnal disinhibition would overstimulate the limbic system and the hippocampus would become overactivated, the metabolic rate at this level being inversely related to P300 amplitude. Clozapine showed a more selective and intense action on that hyperactive metabolic tone than haloperidol. PMID- 12754161 TI - Neonatal exposure to the glutamate receptor antagonist MK-801: effects on locomotor activity and pre-pulse inhibition before and after sexual maturity in rats. AB - Neonatal lesions of the ventral hippocampus in rats lead to post- but not pre pubertal behavioral changes suggesting adolescent onset of dopaminergic hypersensitivity and providing an animal model of schizophrenia. Neonatal exposure to glutamate receptor antagonists produces accelerated apoptosis leading to neuronal loss in central nervous system structures including the hippocampus. This suggested that neonatal MK-801 might lead to behavioural changes like those reported following ventral hippocampal lesions. Thus, rats received MK-801 (0, 0.5, 1.0 mg/kg ip) on postnatal day 3 (P3) and were tested pre- (P35) and post pubertally (P56). MK-801 produced an increase in TUNEL staining in the hippocampus and other forebrain structures, confirming the induction of apoptosis. Results showed little difference in locomotor activity between neonatal saline- and MK-801-treated groups during habituation or following saline injection but increased activity was seen in the 0.5 mg/kg MK-801 group following amphetamine (1.5 mg/kg i.p.) at P35 but not P56. In tests of pre-pulse inhibition (PPI), neonatal saline and MK-801 groups showed stable startle amplitudes, minimal responding to the pre-pulse stimuli alone, an increase in PPI with increases in pre-pulse intensity, and reduced PPI with apomorphine (0.1 mg/kg s.c.). At P56, neonatal MK-801 groups tested following vehicle showed less sensitivity to changes in pre-pulse intensity. It was concluded that neonatal MK 801 increases apoptotic cell loss in the hippocampus but does not produce behavioural effects like those seen after neonatal ventral hippocampal lesions. However, neonatal MK-801 did lead to increases in locomotor activity in juveniles but not adults and reduced sensitivity to pre-pulse intensity in PPI tests in adulthood. PMID- 12754160 TI - A neurodevelopmental model of schizophrenia: neonatal disconnection of the hippocampus. AB - In the context of our current knowledge about schizophrenia, heuristic models of psychiatric disorders may be used to test the plausibility of theories developed on the basis of new emerging biological findings, explore mechanisms of schizophrenia-like phenomena, and develop potential new treatments. In a series of studies, we have shown that neonatal excitotoxic lesions of the rat ventral hippocampus (VH) may serve as a heuristic model. The model appears to mimic a spectrum of neurobiological and behavioral features of schizophrenia, including functional pathology in presumably critical brain regions interconnected with the hippocampal formation and targeted by antipsychotic drugs - the striatum/nucleus accumbens and the prefrontal cortex, and leads in adolescence or early adulthood to the emergence of abnormalities in a number of dopamine related behaviors. Moreover, our data show that even transient inactivation of the ventral hippocampus during a critical period of development, that produces subtle, if any, anatomical changes in the hippocampus, may be sufficient to disrupt normal maturation of the prefrontal cortex (and perhaps, other interconnected late maturing regions) and trigger behavioral changes similar to those observed in animals with the permanent excitotoxic lesion. These results represent a potential new model of aspects of schizophrenia without a gross anatomical lesion. PMID- 12754163 TI - Functional deficits following neonatal dopamine depletion and isolation housing: circular water maze acquisition under pre-exposure conditions and motor activity. AB - Seven experiments and several behavioural tests were performed to study the effects of housing condition and experimental test conditions upon the behavioural responses and performance of adult rats neonatally treated with 6 hydroxydopamine (6-OHDA, 100 ug intracisternally, i.c.) or with vehicle. Postnatal 6-OHDA induced locomotor and total activity hyperactivity and deficits in navigational learning in a circular swim maze that were blocked by pretreatment with a dopamine (DA) reuptake inhibitor but not a noradrenaline (NA) reuptake inhibitor. Isolation-housing induced deficits in maze learning performance. Grouped housing improved the maze learning performance of 6-OHDA treated rats whereas vehicle treated rats that were isolation housed performed better following latent learning (LL) pre-exposure trials. 6-OHDA treated rats that received both Grouped housing and latent learning trials performed better on the spatial navigation task than those that received Grouped housing but no latent learning or Isolation housing and latent learning. Analysis of habituation quotients indicated marked deficits by 6-OHDA-treated rats suggesting inability to acquire this simple, nonassociative form of learning. Methylphenidate increased all three parameters of motor activity: locomotion, rearing and total activity, in both Isolation-housed and Group-housed rats from 60- to 90- or 120 min post-injection. NDO 008 induced variable and parameter-dependent effects: locomotion was elevated initially in both Isolated and Grouped rats by the compound and then reduced in the Isolated rats only whereas total activity was only elevated initially in the Isolated rats and unaffected in the Grouped rats. Rearing behaviour was reduced markedly, directly post-injection, in the Isolation housed rats. DA, DOPAC and HVA concentrations in the striatum, nucleus accumbens, olfactory tubercle and midbrain were reduced but most markedly in the striatum. 5 Hydroxytryptamine (5-HT) and 5-hydroxyindole acetic acid (5-HIAA) concentrations were elevated in the striatum, nucleus accumbens (not 5-HIAA) and olfactory tubercle. PMID- 12754162 TI - Developmental markers of psychiatric disorders as identified by sensorimotor gating. AB - Prepulse inhibition (PPI) of the acoustic startle reflex is an operational measure of sensorimotor gating that is amenable to cross-species comparisons. Deficits in PPI have been repeatedly reported in patients with schizophrenia or other psychiatric disorders characterized by abnormalities in sensory, cognitive, or motor gating. Because some forms of schizophrenia appear to be attributable to early developmental perturbations, many animal studies have examined the influences of various developmental manipulations on PPI in adulthood. For example, isolation rearing of rats from weaning into adulthood leads to a reorganization of brain circuitry including changes in monoamine systems that modulate PPI. Isolation rearing of rats leads to deficits in PPI that are not evident pre-puberty, are enduring in adulthood, and are developmentally specific, in that isolation of adult rats does not affect PPI. The PPI deficits in isolation-reared rats are reversed by typical or atypical antipsychotic treatments, including raclopride, haloperidol, clozapine, olanzapine, quetiapine, and even the putative antipsychotic M100907. In contrast, other psychoactive drugs, such as chlordiazepoxide, diazepam, prazosin, or amitriptyline, do not normalize PPI in isolation-reared rats. Hence, the isolation-rearing model may help identify novel antipsychotics. Thus, social isolation rearing of rats provides a developmentally specific, non-pharmacological manipulation that leads to deficits in sensorimotor gating that mimic those observed in schizophrenia patients and are responsive to antipsychotic medications. PMID- 12754164 TI - Human genetic variation and mental disorders. AB - Despite extensive research, our knowledge of structural or functional pathology of severe mental disorders such as schizophrenia and major affective disorders is limited. The only etiological factor with reasonably firm foundation is inheritance, as evidenced by family, twin and adoption studies, however, the molecular basis of this heritability is not established yet. In the absence of clear knowledge about the biological substrate for these psychiatric disorders all the genes expressed in the human brain are potential candidate genes. Association studies analyze genetic variants (polymorphisms) in genes with presumed functional significance in the pathophysiology of the disease (candidate genes). These genetic studies compare the frequency of the hypothetical risk variant in people affected by the disease and in healthy control representatives of the same population. Also, simplex families with an affected child (trios) or with discordant affected offspring (quadruplets), can be used in the association studies. This strategy constitutes a tool to identify the small and moderate effects of genes in these complex phenotypes. Likewise, these kinds of studies could be useful to explain the effect of genes in some specific traits and symptoms present in functional psychoses. Some of our own results on serotoninergic system genes and major depression are included in this chapter as an example of case-control association developed studies in mental disorders. PMID- 12754159 TI - Neuropeptides involved in the pathophysiology of schizophrenia and major depression. AB - The present review summarizes the findings on the role of neuropeptides in the pathophysiology of schizophrenia and major depression. Several neuropeptides as vasopressin and endorphins in particular, beta-endorphin and gamma-type endorphins, cholecystokinin (CCK), neurotensin, somatostatin and Neuropeptide Y have been implicated in schizophrenia. During the last decade, however, few attempts to explore the significance of most of these and other neuropeptides in the pathophysiology of the disease or their therapeutic potential are found in the literature. An exception is neurotensin, which exerts neuroleptic-like effects in animal studies, while CSF, brain and blood studies are inconclusive. Things are different in major depression. Here much attention is paid to the endocrine abnormalities found in this disorder in particular the increased activity of the hypothalamic-pituitary-adrenal (HPA) axis. Neuropeptides as corticotropin-releasing hormone (CRH), vasopressin and corticosteroids are implicated in the symptomatology of this disorder. As a consequence much work is going on investigating the influence of CRH and corticosteroid antagonists or inhibitors of the synthesis of corticosteroids as potential therapeutic agents. This review emphasizes the role of vasopressin in the increased activity of the HPA axis in major depression and suggests exploration of the influence of the now available non-peptidergic vasopressin orally active V1 antagonists. PMID- 12754166 TI - Regulation of the hypothalamic-pituitary-adrenal axis in the neonatal rat: the role of maternal behavior. AB - There is a growing literature that indicates that exposure to elevated levels of glucocorticoids can result in long term consequences for the developing brain. In the developing rodent there is a period from about day 4-14 when the adrenal response to stress is either minimal or non-existent thus resulting in stable low levels of circulating glucocorticoids. This has been designated as the stress hypo-responsive period (SHRP). Numerous experiments have demonstrated that maternal factors are critical for the regulation of the pup's hypothalamic pituitarty adrenal (HPA) axis and the maintenance of the SHRP. Following 24 h of maternal deprivation the neonatal rat shows elevated basal levels of corticosterone and exhibits a robust corticosterone and ACTH response to mild stress. Further c-fos mRNA in the paraventricular nucleus is enhanced following stress in deprived pups. At least three aspects of maternal behavior play a role in the regulation of the HPA axis during development. Tactile stimulation appears capable of inhibiting most of the brain-related changes that occur following maternal deprivation. Feeding is essential for maintaining the adrenal unresponsive and reduces the sensitivity of the adrenal to ACTH. Passive contact suppresses the response to stress. In the adult corticotropin-releasing hormone (CRH) is the major neuropeptide that controls pituitary ACTH secretion. In the maternally deprived pup CRH gene transcription is down regulated and arginine vasopressin (AVP) appears to assume the major regulatory hormone that modulates ACTH. These data all indicate that maternal factors are responsible for actively inhibiting the endocrine responses to stress postnatally. Further, maternal deprivation also results in increased cell death in several brain regions. Thus during development most of the peripheral and central stress responsive systems are capable of being activated. However, under conditions of normal dam-pup interactions these responses are mostly suppressed by the dam's behavioral interaction with the pups thus preventing the potential toxic effects of increased secretion of glucocorticoids during critical periods of brain development. PMID- 12754167 TI - Neurodevelopmental influences on the immune system reflecting brain pathology. AB - A number of studies have shown that early life events can affect the development of the nervous system, contributing to particular individual differences in later vulnerability to different forms of psychosocial stress related to the environment and lifestyle. Neuropeptides, chemokines (CKs), neurotrophins (NTs) belong to the chemical microenvironment of the cells of the central nervous system (CNS). This paper reviews research performed in our and other laboratories indicating that mass spectrometry should play a significant role in future studies of the structures of proteins/peptides in neuroscience. These applications include peptide metabolism associated with normal and impaired neurone/immune function. Detailed information about peptide/protein processing in the CNS may be studied by using the lymphocyte as a model reflecting different chemical modifications of peptides/proteins related to various psychosomatic disturbances reflecting disorders of environment and lifestyle. PMID- 12754168 TI - Memory retrieval and its lasting consequences. AB - Many, if not all psychiatric diseases are accompanied by memory disturbances, in particular, the dementias, schizophrenia, and, to an extent, mood disorders. Anxiety and stress, on the other hand, cause important alterations of memory, particularly its retrieval. Here we discuss several new findings on the basic mechanisms of consolidation, retrieval and extinction of a prototype form of episodic memory in the rat: conditioned fear. The findings point the way for investigations on the pathology of these aspects of memory in health and disease. Emphasis is placed on the parallel processing of retrieval in several cortical areas, on the links between retrieval and the onset of extinction, on the fact that extinction involves new learning requiring gene expression, and on the differences between the retrieval of recent or remote long-term memories. PMID- 12754169 TI - Hypersensitivity of pulmonary C fibers induced by adenosine in anesthetized rats. AB - Compelling clinical evidence implicates the potential role of adenosine in development of airway hyperresponsiveness and suggests involvement of pulmonary sensory receptors. This study was carried out to determine the effect of a low dose of adenosine infusion on sensitivity of pulmonary C-fiber afferents in anesthetized open-chest rats. Infusion of adenosine (40 microg x kg-1x min-1 i.v. for 90 s) mildly elevated baseline activity of pulmonary C fibers. However, during adenosine infusion, pulmonary C-fiber responses to chemical stimulants and lung inflation (30 cmH2O tracheal pressure) were markedly potentiated; e.g., the response to right atrial injection of capsaicin (0.25 or 0.5 microg/kg) was increased by more than fivefold (change in fiber activity = 2.64 +/- 0.67 and 16.27 +/- 3.11 impulses/s at control and during adenosine infusion, n = 13, P < 0.05), and this enhanced response returned to control in approximately 10 min. The potentiating effect of adenosine infusion was completely blocked by pretreatment with 8-cyclopentyl-1,3-dipropylxanthine (100 microg/kg), a selective antagonist of the adenosine A1 receptor, but was not affected by 3,7-dimethyl-1 propargylxanthine (1 mg/kg), an A2-receptor antagonist, or 3-ethyl-5-benzyl-2 methyl-4-phenylethynyl-6-phenyl-1,4-(+/-)-dihydropyridine-3,5-dicarboxylate (2 mg/kg), an A3-receptor antagonist. This potentiating effect was also mimicked by N6-cyclopentyladenosine (0.25 microg x kg-1 x min-1 for 90 s), a selective agonist of the adenosine A1 receptor. In conclusion, our results showed that infusion of adenosine significantly elevated the sensitivity of pulmonary C-fiber afferents in rat lungs and that this potentiating effect is likely mediated through activation of the adenosine A1 receptor. PMID- 12754170 TI - A multicompartment model of carboxyhemoglobin and carboxymyoglobin responses to inhalation of carbon monoxide. AB - We have developed a model that predicts the distribution of carbon monoxide (CO) in the body resulting from acute inhalation exposures to CO. The model includes a lung compartment, arterial and venous blood compartments, and muscle and nonmuscle soft tissues with both vascular and nonvascular subcompartments. In the model, CO is allowed to diffuse between the vascular and nonvascular subcompartments of the tissues and to combine with myoglobin in the nonvascular subcompartment of muscle tissue. The oxyhemoglobin dissociation curve is represented by a modified Hill equation whose parameters are functions of the carboxyhemoglobin (HbCO) level. Values for skeletal muscle mass and cardiac output are calculated from prediction formulas based on age, weight, and height of individual subjects. We demonstrate that the model fits data from CO rebreathing studies when diffusion of CO into the muscle compartment is considered. The model also fits responses of HbCO to single or multiple exposures to CO lasting for a few minutes each. In addition, the model reproduces reported differences between arterial and venous HbCO levels and replicates predictions from the Coburn-Forster-Kane equation for CO exposures of a 1- to 83-h duration. In contrast to approaches based on the Coburn-Forster-Kane equation, the present model predicts uptake and distribution of CO in both vascular and tissue compartments during inhalation of either constant or variable levels of CO. PMID- 12754165 TI - Crossroads of corticotropin releasing hormone, corticosteroids and monoamines. About a biological interface between stress and depression. AB - Mental disorders are frequently preceded by stressful events or situations. Depression is a typical case in point. This raises the question, is depression - or possibly better: are certain forms of depression - caused by stress? Can stress be a true pathogenic factor? Phrased differently: can stress destabilize neuronal systems in the central nervous system to such an extent that depressive symptoms are generated? This question is discussed with the corticotrophin releasing hormone (CRH) and MA systems and hypothalamic-pituitary-adrenal (HPA) axis as major foci. The following issues are explored: the effect of antidepressants on corticosteroid receptor gene expression; the behavioral sequellae of CRH administration; CRH disturbances in depression; the impact of early life adversity on the development of the CRH system and on stress reactivity; the interrelationships of stress hormones and monoaminergic (MA ergic) transmission and finally the therapeutic potential of CRH and cortisol antagonists. The available data suggest that CRH overdrive and cortisol overproduction may play a pathogenic role in the occurrence of certain types of depression, directly and/or indirectly, i.e. by induction or exacerbation of disturbances in MA ergic transmission. Stress should, thus, become a major focus of biological depression research. PMID- 12754171 TI - Neurohumoral responses during prolonged exercise in humans. AB - This study examined neurohumoral alterations during prolonged exercise with and without hyperthermia. The cerebral oxygen-to-carbohydrate uptake ratio (O2/CHO = arteriovenous oxygen difference divided by arteriovenous glucose difference plus one-half lactate), the cerebral balances of dopamine, and the metabolic precursor of serotonin, tryptophan, were evaluated in eight endurance-trained subjects during exercise randomized to be with or without hyperthermia. The core temperature stabilized at 37.9 +/- 0.1 degrees C (mean +/- SE) in the control trial, whereas it increased to 39.7 +/- 0.2 degrees C in the hyperthermic trial, with a concomitant increase in perceived exertion (P < 0.05). At rest, the brain had a small release of tryptophan (arteriovenous difference of -1.2 +/- 0.3 micromol/l), whereas a net balance was obtained during the two exercise trials. Both the arterial and jugular venous dopamine levels became elevated during the hyperthermic trial, but the net release from the brain was unchanged. During exercise, the O2/CHO was similar across trials, but, during recovery from the hyperthermic trial, the ratio decreased to 3.8 +/- 0.3 (P < 0.05), whereas it returned to the baseline level of approximately 6 within 5 min after the control trial. The lowering of O2/CHO was established by an increased arteriovenous glucose difference (1.1 +/- 0.1 mmol/l during recovery from hyperthermia vs. 0.7 +/- 0.1 mmol/l in control; P < 0.05). The present findings indicate that the brain has an increased need for carbohydrates during recovery from strenuous exercise, whereas enhanced perception of effort as observed during exercise with hyperthermia was not related to alterations in the cerebral balances of dopamine or tryptophan. PMID- 12754172 TI - Hemodynamic and ventilatory effects of manual respiratory physiotherapy techniques of chest clapping, vibration, and shaking in an animal model. AB - Chest clapping, vibration, and shaking were studied in 10 physiotherapists who applied these techniques on an anesthetized animal model. Hemodynamic variables (such as heart rate, blood pressure, pulmonary artery pressure, and right atrial pressure) were measured during the application of these techniques to verify claims of adverse events. In addition, expired tidal volume and peak expiratory flow rate were measured to ascertain effects of these techniques. Physiotherapists in this study applied chest clapping at a rate of 6.2 +/- 0.9 Hz, vibration at 10.5 +/- 2.3 Hz, and shaking at 6.2 +/- 2.3 Hz. With the use of these rates, esophageal pressure swings of 8.8 +/- 5.0, 0.7 +/- 0.3, and 1.4 +/- 0.7 mmHg resulted from clapping, vibration, and shaking respectively. Variability in rates and "forces" generated by these techniques was <20% in average coefficients of variation. In addition, clinical experience accounted for 76% of the variance in vibration rate (P = 0.001). Cardiopulmonary physiotherapy experience and layers of towel used explained approximately 79% of the variance in clapping force (P = 0.004), whereas age and clinical experience explained >80% of variance in shaking force (P = 0.003). Application of these techniques by physiotherapists was found to have no significant effects on hemodynamic and most ventilatory variables in this study. From this study, we conclude that chest clapping, vibration, and shaking 1). can be consistently performed by physiotherapists; 2). are significantly related to physiotherapists' characteristics, particularly clinical experience; and 3). caused no significant hemodynamic effects. PMID- 12754173 TI - Cardiovascular effects of the respiratory muscle metaboreflexes in dogs: rest and exercise. AB - In awake dogs, lactic acid was injected into the phrenic and deep circumflex iliac arteries to elicit the diaphragm and abdominal muscle metaboreflexes, respectively. At rest, injections into the phrenic or deep circumflex iliac arteries significantly increased mean arterial blood pressure 21 +/- 7% and reduced cardiac output 6 +/- 2% and blood flow to the hindlimbs 20 +/- 9%. Simultaneously, total systemic, hindlimb, and abdominal expiratory muscle vascular conductances were reduced. These cardiovascular responses were not accompanied by significant changes in the amplitude or timing of the diaphragm electromyogram. During treadmill exercise that increased cardiac output, hindlimb blood flow, and vascular conductance 159 +/- 106, 276 +/- 309, and 299 +/- 90% above resting values, lactic acid injected into the phrenic or deep circumflex iliac arteries also elicited pressor responses and reduced hindlimb blood flow and vascular conductance. Adrenergic receptor blockade at rest eliminated the cardiovascular effects of the respiratory muscle metaboreflex. We conclude that the cardiovascular effects of respiratory muscle metaboreflex activation are similar to those previously reported for limb muscles. When activated via metabolite production, the respiratory muscle metaboreflex may contribute to the increased sympathetic tone and redistribution of blood flow during exercise. PMID- 12754174 TI - Effects of high-cholesterol diet and parallel exercise training on the vascular function of rabbit aortas: a time course study. AB - It is plausible to assume that exercise training, when applied early enough, can completely correct atherosclerotic defects. Using rabbit aortic specimens, we examined the effects of chronic exercise and high-cholesterol diet feeding on vascular function for different time periods. Male New Zealand White rabbits were divided into four groups: the normal diet groups with or without exercise training and the high-cholesterol diet groups with or without exercise training. Animals in high-cholesterol diet groups were fed 2% cholesterol rabbit chow for 2, 4, or 6 wk. Those in exercise training groups ran on a treadmill at 0.88 km/h for up to 40 min/day, 5 days/wk for the same period of time as the diet feeding. Thoracic aortas were isolated for functional and immunohistochemical analyses. We found that 1). although high-cholesterol diet feeding (>or=2 wk) elevated serum cholesterol levels and impaired acetylcholine-evoked vasorelaxation, only the latter effect was reversed by exercise training; 2). the effects of diet and exercise on acetylcholine-evoked vasorelaxation were mainly due to altered release of nitric oxide and endothelium-derived hyperpolarizing factor; and 3). diet feeding for 4 or 6 wk caused significant lipid deposition and expression of P-selectin, VCAM-1, monocyte chemoattractant protein-1, and inducible nitric oxide synthase, which were largely reduced by exercise training. In conclusion, parallel exercise training almost completely reverses the early-stage endothelial dysfunction caused by high-cholesterol diet feeding. PMID- 12754175 TI - CYP3A5 genotype predicts renal CYP3A activity and blood pressure in healthy adults. AB - A single-nucleotide polymorphism (A6986G) in the cytochrome p-450 3A5 (CYP3A5) gene distinguishes an expressor (*1) and a reduced-expressor (*3) allele and largely predicts CYP3A5 content in liver and intestine. CYP3A5 is the prevailing CYP3A isoform in kidney. We report that, among renal microsomes from 21 organ donors, those from *1/*3 individuals had at least eightfold higher mean kidney microsomal CYP3A5 content and 18-fold higher mean CYP3A catalytic activity than did those from *3/*3 individuals (P = 0.0001 and P = 0.0137, respectively). We also report significant associations between the A6986G polymorphism and systolic blood pressure (P = 0.0007), mean arterial pressure (P = 0.0075), and creatinine clearance (P = 0.0035) among 25 healthy African-American adults. These associations remained significant when sex, age, and body mass index were taken into account. The mean systolic blood pressure of homozygous CYP3A5 expressors (*1/*1) exceeded that of homozygous nonexpressors (*3/*3) by 19.3 mmHg. We speculate whether a high CYP3A5 expressor allele frequency among African Americans may contribute to a high prevalence of sodium-sensitive hypertension in this population. PMID- 12754176 TI - C-reactive protein correlates with macrophage accumulation in coronary arteries of hypercholesterolemic pigs. AB - A growing body of evidence supports the hypothesis that C-reactive protein (CRP) is a marker of inflammation in coronary artery disease. The purpose of the present study was to test the hypothesis that CRP correlates with macrophage accumulation during the initial stages of coronary vascular disease. Adult male pigs were fed a normal chow (NF) or a high-fat high-cholesterol (HF) diet for 20 wk. After 20 wk, blood was collected for analyses of interleukin-6 (IL-6), CRP, and lipids. After blood collection, the pigs were euthanized and the right coronary arteries (RCA) were harvested and fixed in neutral buffered formalin. Paraffin-embedded sections of RCA were stained immunohistochemically for CRP, scavenger receptor A (SRA), and monocyte chemoattractant protein-1 (MCP-1). All cholesterol fractions were elevated in the HF vs. the NF group (P < 0.05). There was little or no positive staining for CRP, SRA, or MCP-1 in the RCA of NF pigs, but there was extensive staining in lipidladen macrophage foam cells in the HF pigs. Double staining revealed colocalization of CRP with SRA and CRP with MCP-1 in foam cells. Serum IL-6 was below the assay detection limit in all pigs. Serum CRP correlated directly with plasma total cholesterol (R = 0.727, P = 0.041) and accumulation of SRA-positive macrophages (R = 0.938, P < 0.001) in RCA of HF pigs. We conclude that serum CRP correlates with macrophage accumulation and coronary artery disease in hypercholesterolemic pigs. PMID- 12754177 TI - Extended high-frequency partial liquid ventilation in lung injury: gas exchange, injury quantification, and vapor loss. AB - High-frequency oscillatory ventilation with perflubron (PFB) reportedly improves pulmonary mechanics and gas exchange and attenuates lung injury. We explored PFB evaporative loss kinetics, intrapulmonary PFB distribution, and dosing strategies during 15 h of high-frequency oscillation (HFO)-partial liquid ventilation (PLV). After saline lavage lung injury, 15 swine were rescued with high-frequency oscillatory ventilation (n = 5), or in addition received 10 ml/kg PFB delivered to dependent lung [n = 5, PLV-compartmented (PLV(C))] or 10 ml/kg distributed uniformly within the lung [n = 5, PLV(U)]. In the PLV(C) group, PFB vapor loss was replaced. ANOVA revealed an unsustained improvement in oxygenation index in the PLV(U) group (P = 0.04); the reduction in oxygenation index correlated with PFB losses. Although tissue myeloperoxidase activity was reduced globally by HFO PLV (P < 0.01) and regional lung injury scores (lung injury scores) in dependent lung were improved (P = 0.05), global lung injury scores were improved by HFO-PLV (P < 0.05) only in atelectasis, edema, and alveolar distension but not in cumulative score. In our model, markers of inflammation and lung injury were attenuated by HFO-PLV, and it appears that uniform intrapulmonary PFB distribution optimized gas exchange during HFO-PLV; additionally, monitoring PFB evaporative loss appears necessary to stabilize intrapulmonary PFB volume. PMID- 12754178 TI - Neural correlates of voluntary breathing in humans. AB - To investigate the functional neuroanatomy of voluntary respiratory control, blood O2 level-dependent functional magnetic resonance imaging was performed in six healthy right-handed individuals during voluntary hyperpnea. Functional images of the whole brain were acquired during 30-s periods of spontaneous breathing alternated with 30-s periods of isocapnic hyperpnea [spontaneous vs. voluntary: tidal volume = 0.5 +/- 0.01 vs. 1.3 +/- 0.1 (SE) liters and breath duration = 4.0 +/- 0.4 vs. 3.2 +/- 0.4 (SE) s]. For the group, voluntary hyperpnea was associated with significant (P < 0.05, corrected for multiple comparisons) neural activity bilaterally in the primary sensory and motor cortices, supplementary motor area, cerebellum, thalamus, caudate nucleus, and globus pallidum. Significant increases in activity were also identified in the medulla (corrected for multiple comparisons on the basis of a small volume correction for a priori region of interest) in a superior dorsal position (P = 0.012). Activity within the medulla suggests that the brain stem respiratory centers may have a role in mediating the voluntary control of breathing in humans. PMID- 12754179 TI - High-impact exercise strengthens bone in osteopenic ovariectomized rats with the same outcome as Sham rats. AB - The effect of jump exercise on middle-aged osteopenic rats was investigated. Forty-two 9-mo-old female rats were either sham-operated (Sham) or ovariectomized (OVX). Three months after surgery, the rats were divided into the following groups: Sham sedentary, Sham exercised, OVX sedentary, and OVX exercised. Rats in the exercise groups jumped 10 times/day, 5 days/wk, for 8 wk, with a jumping height of 40 cm. Less than 1 min was required for the jump training. After the experiment, the right tibia and femur were dissected, and blood was obtained from each rat. OVX rats were observed to have increased body weights and decreased bone mass in their tibiae and femurs. Jump-exercised rats, on the other hand, had significantly increased tibial bone mass, strength, and cortical areas. The bone mass and strength of OVX exercised rats increased to approximately the same extent as Sham exercised rats, despite estrogen deficiency or osteopenia. Our data suggest that jump exercise has beneficial effects on lower limb bone mass, strength, bone mineral density, and morphometry in middle-aged osteopenic rats, as well as in Sham rats. PMID- 12754180 TI - Maximizing quantitative accuracy of lung airway lumen and wall measures obtained from X-ray CT imaging. AB - To objectively quantify airway geometry from three-dimensional computed tomographic (CT) images, an idealized (circular cross section) airway model is parameterized by airway luminal caliber, wall thickness, and tilt angle. Using a two-dimensional CT slice, an initial guess for the airway center, and the full width-half-maximum principle, we form an estimate of inner and outer airway wall locations. We then fit ellipses to the inner and outer airway walls via a direct least squares fit and use the major and minor axes of the ellipses to estimate the tilt and in-plane rotation angles. Convolving the airway model, initialized with these estimates, with the three-dimensional scanner point-spread function forms the predicted image. The difference between predicted and actual images is minimized by refining the model parameter estimates via a multidimensional, unconstrained, nonlinear minimization routine. When optimization converges, airway model parameters estimate the airway inner and outer radii and tilt angle. Results using a Plexiglas phantom show that tilt angle is estimated to within +/ 4 degrees and both inner and outer radii to within one-half pixel when a "standard" CT reconstruction kernel is used. By opening up the ability to measure airways that are not oriented perpendicular to the scanning plane, this method allows evaluation of a greater sampling of airways in a two-dimensional CT slice than previously possible. In addition, by combining the tilt-angle compensation with the deconvolution method, we provide significant improvement over the previous full-width-half-maximum method for assessing location of the luminal edge but not the outer edge of the airway wall. PMID- 12754181 TI - Effects of dichloroacetate on VO2 and intramuscular 31P metabolite kinetics during high-intensity exercise in humans. AB - Traditional control theories of muscle O2 consumption are based on an "inertial" feedback system operating through features of the ATP splitting (e.g., [ADP] feedback, where brackets denote concentration). More recently, however, it has been suggested that feedforward mechanisms (with respect to ATP utilization) may play an important role by controlling the rate of substrate provision to the electron transport chain. This has been achieved by activation of the pyruvate dehydrogenase complex via dichloroacetate (DCA) infusion before exercise. To investigate these suggestions, six men performed repeated, high-intensity, constant-load quadriceps exercise in the bore of an magnetic resonance spectrometer with each of prior DCA or saline control intravenous infusions. O2 uptake (Vo2) was measured breath by breath (by use of a turbine and mass spectrometer) simultaneously with intramuscular phosphocreatine (PCr) concentration ([PCr]), [Pi], [ATP], and pH (by 31P-MRS) and arterialized-venous blood sampling. DCA had no effect on the time constant (tau) of either Vo2 increase or PCr breakdown [tauVo2 45.5 +/- 7.9 vs. 44.3 +/- 8.2 s (means +/- SD; control vs. DCA); tauPCr 44.8 +/- 6.6 vs. 46.4 +/- 7.5 s; with 95% confidence intervals averaging < +/-2 s]. DCA, however, resulted in significant (P < 0.05) reductions in 1). end-exercise [lactate] (-1.0 +/- 0.9 mM), intramuscular acidification (pH, +0.08 +/- 0.06 units), and [Pi] (-1.7 +/- 2.1 mM); 2). the amplitude of the fundamental components for [PCr] (-1.9 +/- 1.6 mM) and Vo2 (-0.1 +/- 0.07 l/min, or 8%); and 3). the amplitude of the Vo2 slow component. Thus, although the DCA infusion lessened the buildup of potential fatigue metabolites and reduced both the aerobic and anaerobic components of the energy transfer during exercise, it did not enhance either tauVo2 or tau[PCr], suggesting that feedback, rather than feedforward, control mechanisms dominate during high intensity exercise. PMID- 12754182 TI - Soil and plant water relations determine photosynthetic responses of C3 and C4 grasses in a semi-arid ecosystem under elevated CO2. AB - To model the effect of increasing atmospheric CO2 on semi-arid grasslands, the gas exchange responses of leaves to seasonal changes in soil water, and how they are modified by CO2, must be understood for C3 and C4 species that grow in the same area. In this study, open-top chambers were used to investigate the photosynthetic and stomatal responses of Pascopyrum smithii (C3) and Bouteloua gracilis (C4) grown at 360 (ambient CO2) and 720 micro mol mol-1 CO2 (elevated CO2) in a semi-arid shortgrass steppe. Assimilation rate (A) and stomatal conductance (gs) at the treatment CO2 concentrations and at a range of intercellular CO2 concentrations and leaf water potentials (psileaf) were measured over 4 years with variable soil water content caused by season and CO2 treatment. Carboxylation efficiency of ribulose bisphosphate carboxylase/oxygenase (Vc,max), and ribulose bisphosphate regeneration capacity (Jmax) were reduced in P. smithii grown in elevated CO2, to the degree that A was similar in elevated and ambient CO2 (when soil moisture was adequate). Photosynthetic capacity was not reduced in B. gracilis under elevated CO2, but A was nearly saturated at ambient CO2. There were no stomatal adaptations independent of photosynthetic acclimation. Although photosynthetic capacity was reduced in P. smithii growing in elevated CO2, reduced gs and transpiration improved soil water content and psileaf in the elevated CO2 chambers, thereby improving A of both species during dry periods. These results suggest that photosynthetic responses of C3 and C4 grasses in this semi-arid ecosystem will be driven primarily by the effect of elevated CO2 on plant and soil water relations. PMID- 12754184 TI - Dexamethasone stimulates transcription of the Na+-K+-ATPase beta1 gene in adult rat lung epithelial cells. AB - Na+-K+-ATPase plays an essential role in active alveolar epithelial fluid resorption. In fetal and adult alveolar epithelial cells, glucocorticoids (GC) increase Na+-K+-ATPase activity and mRNA levels. We sought to define the mechanism of Na+-K+-ATPase gene upregulation by GC. In a rat alveolar epithelial cell line (RLE), dexamethasone (Dex) increased beta1-subunit Na+-K+-ATPase mRNA expression two- to threefold within 3 h after exposure to the GC. The increased gene expression was due to increased transcription as demonstrated by nuclear run on assays, whereas mRNA stability remained unchanged. Transient transfection of 5' deletion mutants of a beta1 promoter-reporter construct demonstrated a 1.5- to 2.2-fold increase in promoter activity by Dex. All of the 5' deletion constructs contained partial or palindromic GC regulatory elements (GRE) and responded to GC. The increased expression of promoter reporter was inhibited by RU-486, a GC receptor (GR) antagonist, suggesting the involvement of GR. The palindromic GRE at -631 demonstrated Dex induction in a heterologous promoter construct. Gel mobility shift assays using RLE nuclear extracts demonstrated specific binding to this site and the presence of GR. We conclude that GC directly stimulate transcription of Na+-K+-ATPase beta1 gene expression in adult rat lung epithelial cells through a GR-dependent mechanism that can act at multiple sites. PMID- 12754183 TI - Lung endothelial heparan sulfates mediate cationic peptide-induced barrier dysfunction: a new role for the glycocalyx. AB - The endothelial glycocalyx is believed to play a major role in microvascular permeability. We tested the hypothesis that specific components of the glycocalyx, via cytoskeletal-mediated signaling, actively participate in barrier regulation. With the use of polymers of arginine and lysine as a model of neutrophil-derived inflammatory cationic proteins, we determined size- and dose dependent responses of cultured bovine lung microvascular endothelial cell permeability as assessed by transendothelial electrical resistance (TER). Polymers of arginine and lysine >11 kDa produced maximal barrier dysfunction as demonstrated by a 70% decrease in TER. Monomers of l-arginine and l-lysine did not alter barrier function, suggesting a cross-linking requirement of cell surface "receptors". To test the hypothesis that glycosaminoglycans (GAGs) are candidate receptors for this response, we used highly selective enzymes to remove specific GAGs before polyarginine (PA) treatment and examined the effect on TER. Heparinase III attenuated PA-induced barrier dysfunction by 50%, whereas heparinase I had no effect. To link changes in barrier function with structural alterations, we examined actin organization and syndecan localization after PA. PA induced actin stress fiber formation and clustering of syndecan-1 and syndecan 4, which were significantly attenuated by heparinase III. PA-induced cytoskeletal rearrangement and barrier function did not involve myosin light chain kinase (MLCK) or p38 MAPK, as ML-7, a specific MLCK inhibitor, or SB-20358, a p38 MAPK inhibitor, did not alter PA-induced barrier dysfunction. In summary, lung endothelial cell heparan sulfate proteoglycans are key participants in inflammatory cationic peptide-induced signaling that links cytoskeletal reorganization with subsequent barrier dysfunction. PMID- 12754185 TI - Differential effects of mechanical ventilatory strategy on lung injury and systemic organ inflammation in mice. AB - Patients with acute respiratory distress syndrome are at increased risk for developing multiorgan system dysfunction. The goal of this study was to establish an in vivo murine model to assess the differential effects of ventilation protective strategies on the development of acute lung injury and systemic organ inflammation. C57B/6 mice were randomized to mechanical ventilation (MV) with conventional, high (17 ml/kg) or protective, low (6 ml/kg) tidal volume (VT) after intratracheal hydrochloric acid or no intervention. Mean arterial pressure was continuously monitored during MV and did not differ between groups. After 4 h, lung injury was assessed by measurement of wet/dry lung weight, lung lavage protein concentration and cell count, and histology. Concentration of IL-6, TNF alpha, VEGF, and VEGF receptor-2 (VEGFR2) was measured in lung, liver, kidney, and heart. Results were compared with control, spontaneously breathing mice. Lung injury and altered pulmonary cytokine expression were not detected after MV of healthy mice with low or high VT. Although MV did not significantly alter IL-6 or TNF-alpha in systemic organs, VEGF concentration significantly increased in liver and kidney. After acid aspiration, mice ventilated with high VT manifested lung injury and increased IL-6 and VEGFR2 in lung, liver, and kidney, whereas VEGF increased only in liver and kidney. MV with low VT after acid aspiration attenuated lung injury, both IL-6 and VEGFR2 expression in lung and systemic organs, and hepatic, but not renal, increased VEGF. Our data suggest that MV strategy has differential effects on systemic inflammatory changes and thus may selectively predispose to systemic organ dysfunction. PMID- 12754186 TI - Hypoxia-induced pulmonary artery adventitial remodeling and neovascularization: contribution of progenitor cells. AB - Information is rapidly emerging regarding the important role of the arterial vasa vasorum in a variety of systemic vascular diseases. In addition, increasing evidence suggests that progenitor cells of bone marrow (BM) origin may contribute to postnatal neovascularization and/or vascular wall thickening that is characteristic in some forms of systemic vascular disease. Little is known regarding postnatal vasa formation and the role of BM-derived progenitor cells in the setting of pulmonary hypertension (PH). We sought to determine the effects of chronic hypoxia on the density of vasa vasorum in the pulmonary artery and to evaluate if BM-derived progenitor cells contribute to the increased vessel wall mass in a bovine model of hypoxia-induced PH. Quantitative morphometric analyses of lung tissue from normoxic and hypoxic calves revealed that hypoxia results in a dramatic expansion of the pulmonary artery adventitial vasa vasorum. Flow cytometric analysis demonstrated that cells expressing the transmembrane tyrosine kinase receptor for stem cell factor, c-kit, are mobilized from the BM in the circulation in response to hypoxia. Immunohistochemistry revealed an increase in the expression of c-kit+ cells together with vascular endothelial growth factor, fibronectin, and thrombin in the hypoxia-induced remodeled pulmonary artery vessel wall. Circulating mononuclear cells isolated from neonatal calves exposed to hypoxia were found to differentiate into endothelial and smooth muscle cell phenotypes depending on culture conditions. From these observations, we suggest that the vasa vasorum and circulating progenitor cells could be involved in vessel wall thickening in the setting of hypoxia-induced PH. PMID- 12754187 TI - Angiotensin II and the fibroproliferative response to acute lung injury. AB - Angiotensin II (ANG II), generated by activation of local renin-angiotensin systems, is believed to play an important role in tissue repair and remodeling, in part via transforming growth factor-beta (TGF-beta). Angiotensin-converting enzyme (ACE) inhibitors have been shown to abrogate experimental lung injury via a number of potential mechanisms; however, the potentially fibroproliferative role for ANG II in the lung has not been characterized. We hypothesized that, after lung injury, ANG II would stimulate fibroblast procollagen synthesis and promote lung collagen deposition in rats. In vitro, ANG II was a potent inducer of procollagen production in human lung fibroblasts via activation of the type 1 receptor and, at least in part, via the autocrine action of TGF-beta. After bleomycin-induced lung injury, an increase in lung ANG II concentration was observed by day 3 that preceded increases in lung collagen and was maintained until death at day 21. Administration of an ACE inhibitor (ramipril) reduced ACE activity, ANG II concentration, TGF-beta expression, and collagen deposition. Losartan (an ANG II type 1 receptor antagonist) also attenuated the increase in TGF-beta expression and lung collagen deposition. These observations suggest that ANG II, possibly generated locally within the lung, may play an important role in the fibrotic response to acute lung injury, at least in part via the action of TGF-beta. ACE inhibitors and receptor antagonists, already widely used clinically, should be assessed as potential new therapies for fibrotic lung disease. PMID- 12754188 TI - Postreceptor defects in alveolar epithelial beta-adrenergic signaling after prolonged isoproterenol infusion. AB - We previously found that prolonged isoproterenol (Iso) infusion in rats impaired the ability of beta-adrenoceptor (beta-AR) agonists to increase alveolar liquid clearance (ALC). Here, we determined if postreceptor defects in beta-AR signaling contribute to this impairment. Iso was infused using subcutaneous miniosmotic pumps (4, 40, or 400 microg. kg-1. h-1) in rats for 48 h. At this time, forskolin stimulated ALC was measured by mass balance. Forskolin-stimulated ALC [33.4 +/- 2.1%/h (mean +/- SE) in vehicle-infused rats] was reduced by 25 and 38%, respectively, after the 40 and 400 microg. kg-1. h-1 Iso infusions. The ability of forskolin to increase cAMP was reduced by 70% in alveolar type II (ATII) cells isolated from rats infused with 400 microg. kg-1. h-1 Iso. Additionally, the ability of the stable cAMP analog 8-bromoadenosine-3',5'-cyclic monophosphorothioate, Sp-isomer, to increase ALC (48.7 +/- 3.0% in vehicle infused rats) was reduced by 25 and 51%, respectively, after the 40 and 400 microg. kg-1. h-1 infusions. Finally, the ability of cAMP to increase protein kinase A activity was eliminated in ATII cells isolated from rats infused with Iso at 400 microg. kg-1. h-1. These data demonstrate that prolonged beta-AR agonist exposure can impair alveolar epithelial beta-AR signaling downstream of the beta-AR. PMID- 12754189 TI - Systemic arteriovenous fistula leads to pulmonary artery remodeling and abnormal vasoreactivity in the fetal lamb. AB - Several cases of systemic arteriovenous fistula diagnosed in the human fetus have been associated with the postnatal development of persistent pulmonary hypertension. The aim of this study was to determine the effects of a prenatally created systemic arteriovenous fistula on the structure and reactivity of the pulmonary circulation in the fetal lamb. A fistula between the jugular vein and carotid artery was created in fetal lambs at 119-124 days of gestation. At delivery (134-139 days), left pulmonary artery (LPA) pressure was increased in the fistula group (n = 12) compared with controls (n = 11, P < 0.01). The pulmonary vascular resistance was significantly higher in the fistula group (P < 0.05), whereas mean LPA blood flow was not statistically different between the two groups. Morphometric analysis of the pulmonary vascular bed revealed an increase in the number of peripheral muscular arteries, together with an increase in pulmonary arterial medial thickness in the fistula group. There was no difference in the relative number or size of intraacinar arteries. In vitro organ bath studies on pulmonary arterial rings showed impaired endothelium-dependent relaxation in the fistula group compared with controls. However, endothelial nitric oxide synthase protein expression was similar in both groups, whereas endothelium-independent relaxation to sodium nitroprusside was greater in the fistula group compared with controls. A systemic arteriovenous fistula leads to both structural and functional alteration of the pulmonary vasculature, which might lead to the development of persistent pulmonary hypertension after birth. PMID- 12754190 TI - 20-HETE inotropic effects involve the activation of a nonselective cationic current in airway smooth muscle. AB - 20-Hydroxyeicosatetraenoic acid (20-HETE) controls several mechanisms such as vasoactivity, mitogenicity, and ion transport in various tissues. Our goal was to quantify the effects of 20-HETE on the electrophysiological properties of airway smooth muscle (ASM). Isometric tension measurements, performed on guinea pig ASM, showed that 20-HETE induced a dose-dependent inotropic effect with an EC50 value of 1.5 microM. This inotropic response was insensitive to GF-109203X, a PKC inhibitor. The sustained contraction, requiring Ca2+ entry, was partially blocked by either 100 microM Gd3+ or 1 microM nifedipine, revealing the involvement of noncapacitative Ca2+ entry and L-type Ca2+ channels, respectively. Microelectrode measurements showed that 3 microM 20-HETE depolarized the membrane potential in guinea pig ASM by 13 +/- 2mV(n = 7), as did 30 microM 1-oleoyl-2-acetyl-sn glycerol. Depolarizing effects were also observed in the absence of epithelium. Patch-clamp recordings demonstrated that 1 microM 20-HETE activated a nonselective cationic inward current that may be supported by the activation of transient receptor potential channels. The presence of canonical transient receptor potential mRNA was confirmed by RT-PCR in guinea pig ASM cells. PMID- 12754191 TI - Effect of oxygen on cyclic GMP-dependent protein kinase-mediated relaxation in ovine fetal pulmonary arteries and veins. AB - Cyclic GMP-dependent protein kinase (PKG) plays an important role in regulating pulmonary vasomotor tone in the perinatal period. In this study, we tested the hypothesis that a change in oxygen tension affects PKG-mediated pulmonary vasodilation. Isolated intrapulmonary arteries and veins of near-term fetal lambs were first incubated for 4 h under hypoxic and normoxic conditions (Po2 of 30 and 140 mmHg, respectively) and then contracted with endothelin-1. 8-Bromoguanosine 3',5'-cyclic monophosphate (8-BrcGMP), a cell membrane-permeable analog of cGMP, induced a greater relaxation in vessels incubated in normoxia than in hypoxia. beta-Phenyl-1,N2-etheno-8-bromoguanosine-3',5'-cyclic monophosphorothioate, Rp isomer (Rp-8-Br-PET-cGMPS), a selective inhibitor of PKG, attenuated relaxation induced by 8-BrcGMP (10-4 and 3 x 10-4 M). In the presence of Rp-8-Br-PET-cGMPS, the differential responses to 8-BrcGMP between hypoxia and normoxia treatment were abolished in veins but not in arteries. cGMP-stimulated PKG activity was present in arteries but not in veins after 4 h of hypoxia. Both vessel types showed significant increase in cGMP-stimulated PKG activity after 4 h of normoxia. PKG protein (Western blot analysis) and PKG mRNA levels (quantitative RT-PCR) were greater in veins but not in arteries after 4-h exposure to normoxia vs. hypoxia. These results demonstrate that oxygen augments cGMP-mediated vasodilation of fetal pulmonary arteries and veins. Furthermore, the effect of oxygen on response of the veins to cGMP is due to an increase in the activity, protein level, and mRNA of PKG. PMID- 12754192 TI - PAR-2 activation, PGE2, and COX-2 in human asthmatic and nonasthmatic airway smooth muscle cells. AB - The protease-activated receptor-2 (PAR-2) is present on human airway smooth muscle (ASM) cells and can be activated by mast cell tryptase, trypsin, or an activating peptide (AP). Trypsin induced significant increases in PGE2 release from human ASM cells after 6 and 24 h and also induced cyclooxygenase (COX)-2 mRNA expression and COX-2 protein. Tryptase and the PAR-2 AP did not alter PGE2 release or COX-2 protein levels, suggesting a lack of PAR-2 involvement. When we compared results in asthmatic and nonasthmatic muscle cells, both trypsin and bradykinin induced less PGE2 from asthmatic ASM cells, and bradykinin induced significantly less COX-2 mRNA in asthmatic cells. Significantly less PGE2 was released from proliferating ASM cells from asthmatic patients. In conclusion, trypsin induces PGE2 release and COX-2 in human ASM cells, which is unlikely to be via PAR-2 activation. In addition, ASM cells from asthmatic patients produce significantly less PGE2 and COX-2 compared with nonasthmatic cells. These findings may contribute to the increase in muscle mass evident in asthmatic airways. PMID- 12754193 TI - Glucocorticoid effects on vitamin K-dependent carboxylase activity and matrix Gla protein expression in rat lung. AB - The role of glucocorticoids in the regulation of vitamin K-dependent carboxylase activity was investigated in fetal and adult lung. Glucocorticoid deficiency induced by adrenalectomy (ADX) stimulated adult lung growth and reduced carboxylation in a tissue-specific manner. Type II epithelial cells were enriched in carboxylase activity, where ADX-induced downregulation was retained in freshly isolated cells. Carboxylase activity in fetal type II cells was one-half that found in fetal fibroblasts isolated from the same lungs, and both populations increased activity with time in culture. Both carboxylase activity and formation of gamma-carboxyglutamate (Gla)-containing proteins were stimulated by dexamethasone (Dex) in fetal type II cells. Matrix Gla protein (MGP), a vitamin K dependent protein known to be synthesized in type II cells, was also found in fetal fibroblasts, where its expression was stimulated by Dex. These combined results suggested an important role for glucocorticoids and MGP in the developing lung, where both epithelial and mesenchymal cells coordinate precise control of branching morphogenesis. We investigated MGP expression and its regulation by Dex in the fetal lung explant model. MGP mRNA and protein were increased in parallel with the formation of highly branched lungs, and this increase was stimulated twofold by Dex at each day of culture. Dex-treated explants were characterized by large, dilated, conducting airways and a peripheral rim of highly branched saccules compared with uniformly branched controls. We propose that glucocorticoids are important regulators of vitamin K function in the developing and adult lung. PMID- 12754194 TI - Protein C and thrombomodulin in human acute lung injury. AB - Decreased circulating protein C and increased circulating thrombomodulin are markers of the prothrombotic, antifibrinolytic state associated with poor outcomes in sepsis but have not been measured in patients with ALI (acute lung injury)/ARDS (acute respiratory distress syndrome). We measured circulating and intra-alveolar protein C and thrombomodulin in 45 patients with ALI/ARDS from septic and nonseptic causes and correlated the levels with clinical outcomes. Plasma protein C levels were lower in ALI/ARDS compared with normal. Lower levels of protein C were associated with worse clinical outcomes, including death, fewer ventilator-free days, and more nonpulmonary organ failures, even when only patients without sepsis were analyzed. Levels of thrombomodulin in pulmonary edema fluid from ALI/ARDS patients were >10-fold higher than normal plasma and 2 fold higher than ALI/ARDS plasma. Higher edema fluid thrombomodulin levels were associated with worse clinical outcomes. The higher levels in edema fluid compared with plasma suggest local release of soluble thrombomodulin in the lung, possibly from a lung epithelial source. To determine whether lung epithelial cells can release thrombomodulin, A549 cells and primary isolates of human alveolar type II cells were exposed to H2O2 or inflammatory cytokines. Both epithelial cell types released thrombomodulin into the media. In summary, the protein C system is markedly disrupted in patients with ALI/ARDS from both septic and nonseptic causes. The protein C system may be a potential therapeutic target in patients with ALI/ARDS. PMID- 12754195 TI - The three-dimensional structure of the core domain of Naf Y from Azotobacter vinelandii determined at 1.8-A resolution. AB - The Azotobacter vinelandii NafY protein (nitrogenase accessory factor Y) is able to bind either to the iron molybdenum cofactor (FeMo-co) or to apodinitrogenase and is believed to facilitate the transfer of FeMo-co into apodinitrogenase. The NafY protein has two domains: an N-terminal domain (residues Met1-Leu98) and a C terminal domain (residues Glu99-Ser232), referred here to as the "core domain." The core domain of NafY is shown here to be capable of binding the FeMo cofactor of nitrogenase but unable to bind to apodinitrogenase in the absence of the first domain. The three-dimensional molecular structure of the core domain of NafY has been solved to 1.8-A resolution, revealing that the protein consists of a mixed five-stranded beta-sheet flanked by five alpha-helices that belongs to the ribonuclease H superfamily. As such, this represents a new fold capable of binding FeMo-co, where the only previous example was that seen in dinitrogenase. PMID- 12754197 TI - Antifungal activity of amiodarone is mediated by disruption of calcium homeostasis. AB - The antiarrhythmic drug amiodarone was recently demonstrated to have novel broad range fungicidal activity. We provide evidence that amiodarone toxicity is mediated by disruption of Ca2+ homeostasis in Saccharomyces cerevisiae. In mutants lacking calcineurin and various Ca2+ transporters, including pumps (Pmr1 and Pmc1), channels (Cch1/Mid1 and Yvc1), and exchangers (Vcx1), amiodarone sensitivity correlates with cytoplasmic calcium overload. Measurements of cytosolic Ca2+ by aequorin luminescence demonstrate a biphasic response to amiodarone. An immediate and extensive calcium influx was observed that was dose dependent and correlated with drug sensitivity. The second phase consisted of a sustained release of calcium from the vacuole via the calcium channel Yvc1 and was independent of extracellular Ca2+ entry. To uncover additional cellular pathways involved in amiodarone sensitivity, we conducted a genome-wide screen of nearly 5000 single-gene yeast deletion mutants. 36 yeast strains with amiodarone hypersensitivity were identified, including mutants in transporters (pmr1, pdr5, and vacuolar H+-ATPase), ergosterol biosynthesis (erg3, erg6, and erg24), intracellular trafficking (vps45 and rcy1), and signaling (ypk1 and ptc1). Of three mutants examined (vps45, vma3, and rcy1), all were found to have defective calcium homeostasis, supporting a correlation with amiodarone hypersensitivity. We show that low doses of amiodarone and an azole (miconazole, fluconazole) are strongly synergistic and exhibit potent fungicidal effects in combination. Our findings point to the potentially effective application of amiodarone as a novel antimycotic, particularly in combination with conventional antifungals. PMID- 12754198 TI - Importance of nuclear localization of apoptin for tumor-specific induction of apoptosis. AB - The chicken anemia virus-derived protein Apoptin induces apoptosis specifically in human tumor and transformed cells and not in normal, untransformed cells. The cell killing activity correlates with a predominantly nuclear localization of Apoptin in tumor cells, whereas in normal cells, it is detected mainly in cytoplasmic structures. To explore the role of nuclear localization for Apoptin induced cell death in tumor cells, we employed a mutagenesis strategy. First, we demonstrated that the C terminus of Apoptin contains a bipartite-type nuclear localization signal. Strikingly, further investigation showed that Apoptin contains two different domains that induce apoptosis independently, and for both domains, we found a strong correlation between localization and killing activity. Using inhibitors, we ruled out the involvement of de novo gene transcription and translation and further showed that Apoptin itself does not have any significant transcriptional repression activity, suggesting that Apoptin exerts its effects in the nucleus by some other method. To determine whether nuclear localization is sufficient to enable Apoptin to kill normal, untransformed cells, we expressed full-length Apoptin fused to a heterologous nuclear localization signal in these cells. However, despite its nuclear localization, no apoptosis was induced, which suggests that nuclear localization per se is not sufficient for Apoptin to become active. These studies increase our understanding of the molecular pathway of Apoptin and may also shed light on the mechanism of cellular transformation. PMID- 12754199 TI - Synapsin I-associated phosphatidylinositol 3-kinase mediates synaptic vesicle delivery to the readily releasable pool. AB - Maintaining synaptic transmission requires replenishment of docked synaptic vesicles within the readily releasable pool (RRP) from synaptic vesicle clusters in the synapsin-bound reserve pool. We show that synapsin forms a complex with phosphatidylinositol 3-kinase (PI 3-kinase) in intact nerve terminals and that synapsin-associated kinase activity increases on depolarization. Disruption of either PI 3-kinase activity or its interaction with synapsin inhibited replenishment of the RRP, but did not affect exocytosis from the RRP. Thus we conclude that a synapsin-associated PI 3-kinase activity plays a role in synaptic vesicle delivery to the RRP. This also suggests that PI 3-kinase contributes to the maintenance of synaptic transmission during periods of high activity, indicating a possible role in synaptic plasticity. PMID- 12754200 TI - Farnesoid X receptor regulates bile acid-amino acid conjugation. AB - The farnesoid X receptor (FXR; NR1H4) regulates bile acid and lipid homeostasis by acting as an intracellular bile acid-sensing transcription factor. Several identified FXR target genes serve critical roles in the synthesis and transport of bile acids as well as in lipid metabolism. Here we used Affymetrix micro-array and Northern analysis to demonstrate that two enzymes involved in conjugation of bile acids to taurine and glycine, namely bile acid-CoA synthetase (BACS) and bile acid-CoA: amino acid N-acetyltransferase (BAT) are induced by FXR in rat liver. Analysis of the human BACS and BAT genes revealed the presence of functional response elements in the proximal promoter of BACS and in the intronic region between exons 1 and 2 of the BAT gene. The response elements resemble the consensus FXR binding site consisting of two nuclear receptor half-sites organized as an inverted repeat and separated by a single nucleotide (IR-1). These response elements directly bind FXR/retinoid X receptor (RXR) heterodimers and confer the activity of FXR ligands in transient transfection experiments. Further mutational analysis confirms that the IR-1 sequence of the BACS and BAT genes mediate transactivation by FXR/RXR heterodimers. Finally, Fisher rats treated with the synthetic FXR ligand GW4064 clearly show increased transcript levels of both the BACS and BAT mRNA. These studies demonstrate a mechanism by which FXR regulates bile acid amidation, a critical component of the enterohepatic circulation of bile acids. PMID- 12754201 TI - Expression of liver X receptor target genes decreases cellular amyloid beta peptide secretion. AB - A hallmark of Alzheimer's disease is the deposition of plaques of amyloid beta peptide (Abeta) in the brain. Abeta is thought to be formed from the amyloid precursor protein (APP) in cholesterol-enriched membrane rafts, and cellular cholesterol depletion decreases Abeta formation. The liver X receptors (LXR) play a key role in regulating genes that control cellular cholesterol efflux and membrane composition and are widely expressed in cells of the central nervous system. We show that treatment of APP-expressing cells with LXR activators reduces the formation of Abeta. LXR activation resulted in increased levels of the ATP-binding cassette transporter A1 (ABCA1) and stearoyl CoA desaturase, and expression of these genes individually decreased formation of Abeta. Expression of ABCA1 led to both decreased beta-cleavage product of APPSw (i.e. C99 peptide) and reduced gamma-secretase-cleavage of C99 peptide. Remarkably, these effects of ABCA1 on APP processing were independent of cellular lipid efflux. LXR and ABCA1 induced changes in membrane lipid organization had favorable effects on processing of APP, suggesting a new approach to the treatment of Alzheimer's disease. PMID- 12754202 TI - Sodium/calcium exchanger (NCX1) macromolecular complex. AB - The sodium-calcium exchanger, NCX1, is a ubiquitously expressed membrane protein essential in calcium homeostasis for many cells including those in mammalian heart and brain. The function of NCX1 depends on subcellular ("local") factors, the phosphorylation state of NCX1, and the subcellular location of NCX1 within the cell. Here we investigate the molecular organization of NCX1 within the cardiac myocyte. We show that NCX1 is dynamically phosphorylated by protein kinase A (PKA)-dependent phosphorylation in vitro. We also provide evidence that the regulation of this phosphorylation is attributed to the existence of an NCX1 macromolecular complex. Specifically, we show that the macromolecular complex includes both the catalytic and regulatory subunits of PKA. However, only the RI regulatory subunit is found in this macromolecular complex, not RII. Other critical regulatory enzymes are also associated with NCX1, including protein kinase C (PKC) and two serine/threonine protein phosphatases, PP1 and PP2A. Importantly, the protein kinase A-anchoring protein, mAKAP, is found and its presence in the macromolecular complex suggests that these regulatory enzymes are coordinately positioned to regulate NCX1 as has been found in diverse cells for a number of channel proteins. Dual immunocytochemical staining showed the colocalization of NCX1 protein with mAKAP and PKA-RI proteins in cardiomyocytes. Finally, leucine/isoleucine zipper motifs have been identified as possible sites of interaction. Our finding of an NCX1 macromolecular complex in heart suggests how NCX1 regulation is achieved in heart and other cells. The existence of the NCX1 macromolecular complex may also provide an explanation for recent controversial findings. PMID- 12754204 TI - Dysregulated ryanodine receptors mediate cellular toxicity: restoration of normal phenotype by FKBP12.6. AB - Ca2+ homeostasis is a vital cellular control mechanism in which Ca2+ release from intracellular stores plays a central role. Ryanodine receptor (RyR)-mediated Ca2+ release is a key modulator of Ca2+ homeostasis, and the defective regulation of RyR is pathogenic. However, the molecular events underlying RyR-mediated pathology remain undefined. Cells stably expressing recombinant human RyR2 (Chinese hamster ovary cells, CHOhRyR2) had similar resting cytoplasmic Ca2+ levels ([Ca2+]c) to wild-type CHO cells (CHOWT) but exhibited increased cytoplasmic Ca2+ flux associated with decreased cell viability and proliferation. Intracellular Ca2+ flux increased with human RyR2 (hRyR2) expression levels and determined the extent of phenotypic modulation. Co-expression of FKBP12.6, but not FKBP12, or incubation of cells with ryanodine suppressed intracellular Ca2+ flux and restored normal cell viability and proliferation. Restoration of normal phenotype was independent of the status of resting [Ca2+]c or ER Ca2+ load. Heparin inhibition of endogenous inositol trisphosphate receptors (IP3R) had little effect on intracellular Ca2+ handling or viability. However, purinergic stimulation of endogenous IP3R resulted in apoptotic cell death mediated by hRyR2 suggesting functional interaction occurred between IP3R and hRyR2 Ca2+ release channels. These data demonstrate that defective regulation of RyR causes altered cellular phenotype via profound perturbations in intracellular Ca2+ signaling and highlight a key modulatory role of FKBP12.6 in hRyR2 Ca2+ channel function. PMID- 12754203 TI - Csm, a cardiac-specific isoform of the RNA helicase Mov10l1, is regulated by Nkx2.5 in embryonic heart. AB - Nkx2.5 (also called Csx) is an evolutionarily conserved cardiac transcription factor of the homeobox gene family. Nkx2.5 is required for early heart development, because Nkx2.5 null mice die before completion of cardiac looping. To identify genes regulated by Nkx2.5 in the developing heart, we performed differential screening in combination with suppression subtractive hybridization using RNA isolated from wild-type and Nkx2.5 null hearts at embryonic day 8.5. One gene that we found to be markedly down-regulated in the hearts from Nkx2.5 null embryos is an isoform of Mov10 like-1 (Mov10l1), a putative RNA helicase expressed in testis. We named this novel isoform as Csm (cardiac-specific isoform of Mov10l1). Csm is identical with the 3' region of the Mov10l1 gene, but its transcript starts from the exon 16 of Mov10l1. The conceptual protein encoded by Csm cDNA contains a helicase motif as well as ATPase and RNA interaction motifs. Csm is expressed specifically in the heart, and its expression in the heart is restricted to cardiac myocytes. Csm potentiated phenylephrine-induced hypertrophic response in cardiac myocytes. Furthermore, transient cotransfection analysis showed that Nkx2.5 transactivates the Csm promoter, suggesting that Nkx2.5 is essential for embryonic Csm expression. PMID- 12754205 TI - Transcriptional regulation of tristetraprolin by transforming growth factor-beta in human T cells. AB - Transforming growth factor-beta (TGF-beta) is a pleiotropic cytokine that plays a critical role in modulating immune response and inflammation. We employed the Affymetrix cDNA microarray system to detect genes whose expression is regulated by TGF-beta1 in a human T cell line HuT78. Tristetraprolin (TTP), a protein involved in the degradation of tumor necrosis factor-alpha (TNF-alpha) mRNA, was found to be up-regulated by TGF-beta. This up-regulation was confirmed by reverse transcriptase-PCR analysis that revealed a rapid and transient induction of TTP mRNA by TGF-beta 1 in HuT78 cells, primary human T cells, and THP-1 macrophage monocyte cells. In addition, de novo protein synthesis was not required for this induction, suggesting that TTP is regulated by TGF-beta at the transcriptional level. To delineate the transcriptional regulation of the TTP gene, a 2.7-kb human TTP promoter region (-2682 to +56 bp relative to the transcription initiation site) was isolated. We found that this promoter was stimulated by TGF beta 1 or a constitutively active TGF-beta type I receptor via TGF-beta-specific Smad proteins. Furthermore, a series of TTP promoter deletion constructs were used to localize the Smad-responsive region to the -583 to -263 bp portion of the promoter. In this region, the TTP promoter contained a stretch of putative Smad binding elements that had a synergistic effect in mediating Smad activation of the promoter. These putative Smad-binding element-containing sequences were also able to bind Smad3 and Smad4 proteins purified in vitro. As TGF-beta- and TTP deficient mice exhibit overlapping phenotypes manifested by multifocal inflammation and autoimmunity, our findings that TTP transcription is under the control of TGF-beta signaling would indicate a potential role of TTP in mediating the immune suppressive action of TGF-beta in vivo. PMID- 12754206 TI - Localization and characterization of the inhibitory Ca2+-binding site of Physarum polycephalum myosin II. AB - A myosin II is thought to be the driving force of the fast cytoplasmic streaming in the plasmodium of Physarum polycephalum. This regulated myosin, unique among conventional myosins, is inhibited by direct Ca2+ binding. Here we report that Ca2+ binds to the first EF-hand of the essential light chain (ELC) subunit of Physarum myosin. Flow dialysis experiments of wild-type and mutant light chains and the regulatory domain revealed a single binding site that shows moderate specificity for Ca2+. The regulatory light chain, in contrast to regulatory light chains of higher eukaryotes, is unable to bind divalent cations. Although the Ca2+-binding loop of ELC has a canonical sequence, replacement of glutamic acid to alanine in the -z coordinating position only slightly decreased the Ca2+ affinity of the site, suggesting that the Ca2+ coordination is different from classical EF-hands; namely, the specific "closed-to-open" conformational transition does not occur in the ELC in response to Ca2+. Ca2+- and Mg2+ dependent conformational changes in the microenvironment of the binding site were detected by fluorescence experiments. Transient kinetic experiments showed that the displacement of Mg2+ by Ca2+ is faster than the change in direction of cytoplasmic streaming; therefore, we conclude that Ca2+ inhibition could operate in physiological conditions. By comparing the Physarum Ca2+ site with the well studied Ca2+ switch of scallop myosin, we surmise that despite the opposite effect of Ca2+ binding on the motor activity, the two conventional myosins could have a common structural basis for Ca2+ regulation. PMID- 12754207 TI - Critical role of integrin alpha 5 beta 1 in urokinase (uPA)/urokinase receptor (uPAR, CD87) signaling. AB - Urokinase-type plasminogen activator (uPA) induces cell adhesion and chemotactic movement. uPA signaling requires its binding to uPA receptor (uPAR/CD87), but how glycosylphosphatidylinositol-anchored uPAR mediates signaling is unclear. uPAR is a ligand for several integrins (e.g. alpha 5 beta 1) and supports cell-cell interaction by binding to integrins on apposing cells (in trans). We studied whether binding of uPAR to alpha 5 beta 1 in cis is involved in adhesion and migration of Chinese hamster ovary cells in response to immobilized uPA. This process was temperature-sensitive and required mitogen-activated protein kinase activation. Anti-uPAR antibody or depletion of uPAR blocked, whereas overexpression of uPAR enhanced, cell adhesion to uPA. Adhesion to uPA was also blocked by deletion of the growth factor domain (GFD) of uPA and by anti-GFD antibody, whereas neither the isolated uPA kringle nor serine protease domain supported adhesion directly. Interestingly, anti-alpha 5 antibody, RGD peptide, and function-blocking mutations in alpha 5 beta 1 blocked adhesion to uPA. uPA induced cell migration also required GFD, uPAR, and alpha 5 beta 1, but alpha 5 beta 1 alone did not support uPA-induced adhesion and migration. Thus, binding of uPA causes uPAR to act as a ligand for alpha 5 beta 1 to induce cell adhesion, intracellular signaling, and cell migration. We demonstrated that uPA induced RGD dependent binding of uPAR to alpha 5 beta 1 in solution. These results suggest that uPA-induced adhesion and migration of Chinese hamster ovary cells occurs as a consequence of (a) uPA binding to uPAR through GFD, (b) the subsequent binding of a uPA.uPAR complex to alpha 5 beta 1 via uPAR, and (c) signal transduction through alpha 5 beta 1. PMID- 12754208 TI - Thermoirreversible and thermoreversible promoter opening by two Escherichia coli RNA polymerase holoenzymes. AB - Promoter opening, in which the complementary DNA strands separate around the transcriptional start site, is generally thermoreversible. An exceptional case of thermoirreversible opening of the T4 late promoter has been analyzed by KMnO4 footprinting and transcription. T4 late promoters, which consist of an 8-base pair (bp) TATA box "-10" element, are recognized by the small, phage-encoded, highly diverged sigma-family initiation subunit gp55. The T4 late promoter only opens above 15-20 degrees C, but once it has been formed remains open and transcriptionally active for days at -0.5 degrees C. The low temperature-trapped open complex and its isothermally formed state are shown to be structurally distinctive. Two "extended -10" sigma 70 promoters, which, like the T4 late promoter, lack "-35" sites, have been subjected to a comparative analysis: the T4 middle promoter PrIIB2 opens and closes thermoreversibly under conditions of basal and MotA- and AsiA-activated transcription. The open galP1 promoter complex, whose transcription bubble is very AT-rich, also closes reversibly upon shift to -0.5 degrees C, but more slowly than does the rIIB2 promoter. Formation of a trapped-open low temperature state of the promoter complex appears to be a singular property of gp55-RNA polymerase holoenzyme. PMID- 12754209 TI - A bipartite mechanism for ERK2 recognition by its cognate regulators and substrates. AB - Mitogen-activated protein (MAP) kinases control gene expression in response to extracellular stimuli and exhibit exquisite specificity for their cognate regulators and substrates. We performed a structure-based mutational analysis of ERK2 to identify surface areas that are important for recognition of its interacting proteins. We show that binding and activation of MKP3 by ERK2 involve two distinct protein-protein interaction sites in ERK2. Thus, the common docking (CD) site composed of Glu-79, Tyr-126, Arg-133, Asp-160, Tyr-314, Asp-316, and Asp-319 are important for high affinity MKP3 binding but not essential for ERK2 induced MKP3 activation. MKP3 activation requires residues Tyr-111, Thr-116, Leu 119, Lys-149, Arg-189, Trp-190, Glu-218, Arg-223, Lys-229, and His-230 in the ERK2 substrate-binding region, located distal to the common docking site. Interestingly, many of the residues important for MKP3 recognition are also used for Elk1 binding and phosphorylation. In addition to the shared residues, there are also residues that are unique to each target recognition. There is evidence indicating that the CD site and the substrate-binding region defined here are also utilized for MEK1 recognition, and indeed, we demonstrate that the binding of MKP3, Elk1, and MEK1 to ERK2 is mutually exclusive. Taken together, our data suggest that the efficiency and fidelity of ERK2 signaling is achieved by a bipartite recognition process. In this model, one part of the ERK2-binding proteins (e.g. the kinase interaction motif sequence) docks to the CD site located on the back side of the ERK2 catalytic pocket for high affinity association, whereas the interaction of the substrate-binding region with another structural element (e.g. the FXFP motif in MKP3 and Elk1) may not only stabilize binding but also provide contacts crucial for modulating the activity and/or specificity of ERK2 target molecules. PMID- 12754210 TI - The role of Zn2+ in Shal voltage-gated potassium channel formation. AB - Voltage-gated potassium channels are formed by the tetramerization of their alpha subunits, in a process that is controlled by their conserved N-terminal T1 domains. The crystal structures of Shaker and Shaw T1 domains reveal interesting differences in structures that are contained within a highly conserved BTB/POZ domain fold. The most surprising difference is that the Shaw T1 domain contains an intersubunit Zn2+ ion that is lacking in the Shaker T1 domain. The Zn2+ coordination motif is conserved in other non-Shaker channels making this the most distinctive difference between these channels and Shaker. In this study we show that Zn2+ is an important co-factor for the tetramerization of isolated Shaw and Shal T1 domains. Addition of Zn2+ increases the amount of tetramer formed, whereas chelation of Zn2+ with phenanthroline blocks tetramerization and causes assembled tetramers to disassemble. Within an intact cell, full-length Shal subunits containing Zn2+ site mutations also fail to form functional channels, with the majority of the protein found to remain monomeric by size exclusion chromatography. Therefore, zinc-mediated tetramerization also is a physiologically important event for full-length functional channel formation. PMID- 12754211 TI - Protein kinase Calpha-induced p115RhoGEF phosphorylation signals endothelial cytoskeletal rearrangement. AB - Heterotrimeric G-proteins of the Galpha12/13 family activate Rho GTPase through the guanine nucleotide exchange factor p115RhoGEF. Because Rho activation is also dependent on protein kinase Calpha (PKCalpha), we addressed the possibility that PKCalpha can also induce Rho activation secondary to the phosphorylation of p115RhoGEF. Studies were made using human umbilical vein endothelial cells in which we addressed the mechanisms of PKCalpha-induced Rho activation and its consequences on actin cytoskeletal changes. We observed that PKCalpha associated with p115RhoGEF within 1 min of thrombin stimulation and p115RhoGEF phosphorylation was dependent on PKCalpha. Inhibition of PKCalpha-dependent p115RhoGEF phosphorylation prevented the thrombin-induced Rho activation, indicating that the response occurred downstream of PKCalpha phosphorylation of p115RhoGEF. The regulator of G-protein signaling domain of p115RhoGEF, a GTPase activating protein for G12/13, also prevented thrombin-induced Rho activation, indicating the parallel requirement of G12/13 in signaling Rho activation via p115RhoGEF. These data demonstrate a pathway of Rho activation involving PKCalpha dependent phosphorylation of p115RhoGEF. Thus, Rho activation in endothelial cells and the subsequent actin cytoskeletal re-arrangement require the cooperative interaction of both G12/13 and PKCalpha pathways that converge at p115RhoGEF. PMID- 12754212 TI - Identification of small PDZK1-associated protein, DD96/MAP17, as a regulator of PDZK1 and plasma high density lipoprotein levels. AB - Scavenger receptor class B, type I (SR-BI) is the high density lipoprotein (HDL) receptor essential for hepatic uptake of HDL cholesterol. SR-BI was shown to impact plasma HDL levels and be anti-atherogenic. Thus, the ability to regulate hepatic SR-BI may allow for the modulation of plasma HDL cholesterol and progression of atherosclerosis. However, regulation of SR-BI in liver is not well understood. Recently, the PDZ domain containing protein PDZK1 was shown to interact with SR-BI and may serve an essential role in SR-BI cell surface expression. Here we identify an in vivo PDZK1-interacting protein that we named small PDZK1-associated protein (SPAP; also known as DD96/MAP17). Unexpectedly, we found that hepatic overexpression of SPAP in mice resulted in liver deficiency of PDZK1. The absence of PDZK1 in SPAP transgenic mice resulted in a deficiency of SR-BI in liver and markedly increased plasma HDL. Metabolic labeling experiments showed that the proteasome plays a role in the turnover of newly synthesized PDZK1, but that SPAP overexpression in liver increased PDZK1 turnover in an alternate, proteasome-independent pathway. Thus, SPAP may be an endogenous regulator of cellular PDZK1 levels by regulating PDZK1 turnover. PMID- 12754213 TI - Inhibition of interferon (IFN) gamma-induced Jak-STAT1 activation in microglia by vasoactive intestinal peptide: inhibitory effect on CD40, IFN-induced protein-10, and inducible nitric-oxide synthase expression. AB - Interferon (IFN)-gamma is one of the most important microglia stimulators in vivo participating in inflammation and Th1 activation/differentiation. IFN-gamma mediated signaling involves the activation of the Jak/STAT1 pathway. The neuropeptides vasoactive intestinal peptide (VIP) and the pituitary adenylate cyclase activating polypeptide (PACAP) are two potent microglia-deactivating factors that inhibit the production of proinflammatory mediators in vitro and in vivo. The present study investigated the molecular mechanisms involved in the VIP/PACAP regulation of several IFN-gamma-induced microglia-derived factors, including IFN-gamma-inducible protein-10 (IP-10), inducible nitric-oxide synthase (iNOS), and CD40. The results indicate that VIP/PACAP inhibit Jak1-2 and STAT1 phosphorylation, and the binding of activated STAT1 to the IFN-gamma activated site motif in the IFN regulatory factor-1 and CD40 promoter and to the IFN stimulated response element motif of the IP-10 promoter. Through its effect in the IFN-gamma-induced Jak/STAT1 pathway, VIP and PACAP are able to control the gene expression of IP-10, CD40, and iNOS, three microglia-derived mediators that play an essential role in several pathologies, i.e. inflammation and autoimmune disorders. The effects of VIP/PACAP are mediated through the specific receptor VPAC1 and the cAMP/protein kinase A transduction pathway. Because IFN-gamma is a major stimulator of innate and adaptive immune responses in vivo, the down regulation of IFN-gamma-induced gene expression by VIP and PACAP could represent a significant element in the regulation of the inflammatory response in the central nervous system by endogenous neuropeptides. PMID- 12754214 TI - Yin Yang 1 is increased in human heart failure and represses the activity of the human alpha-myosin heavy chain promoter. AB - Yin Yang 1 (YY1) is a transcription factor that can repress or activate transcription of the genes with which it interacts. In this report we show that YY1 is a negative regulator of the alpha-myosin heavy chain (alphaMyHC) gene, which, with betaMyHC are the molecular motors of the heart. AlphaMyHC mRNA and protein levels are down-regulated in hypertrophy and heart failure, and this is thought to be detrimental for cardiac contractility. We show that YY1 specifically interacts with the alphaMyHC promoter and that overexpression of YY1 in cardiac cells represses the activity of the alphaMyHC promoter. We also show that the 170-200-amino acid region of YY1, important for its interaction with histone acetyl transferases and histone deacetylases, is important for its repressive activity and that YY1 deleted in this region is an activator of the alphaMyHC promoter. Moreover, we show that YY1 levels and DNA binding activity are increased in failing human left ventricles and in a mouse model of hypertrophic cardiomyopathy, where alphaMyHC levels are decreased. These results suggest that YY1 is a negative regulator of alphaMyHC gene expression. PMID- 12754215 TI - Lipopolysaccharide (LPS)-binding protein inhibits responses to cell-bound LPS. AB - Lipopolysaccharide (LPS)-binding protein (LBP) is an acute phase reactant that may play a dual role in vivo, both potentiating and decreasing cell responses to bacterial LPS. Whereas low concentrations of LBP potentiate cell stimulation by transferring LPS to CD14, high LBP concentrations inhibit cell responses to LPS. One inhibitory mechanism involves the ability of LBP to neutralize LPS by transferring it to plasma lipoproteins, whereas other inhibitory mechanisms, such as the one described here, do not require exogenous lipoproteins. Here we show that LBP can inhibit monocyte responses to LPS that has already bound to membrane bound CD14 (mCD14) on the cell surface. LBP caused rapid dissociation of LPS from mCD14 as measured by the ability of LBP to inhibit cross-linking of a radioiodinated, photoactivatable LPS derivative to mCD14. Whereas LBP removed up to 75% of the mCD14-bound LPS in 10 min, this was not accompanied by extensive release of the LPS from the cells. The cross-linking data suggest that much of the LPS that remained bound to the cells was associated with LBP. The ability of LBP to inhibit cell responses could not be explained by its effect on LPS internalization, because LBP did not significantly increase the internalization of the cell-bound LPS. In cell-free LPS cross-linking experiments, LBP inhibited the transfer of LPS from soluble CD14 to soluble MD-2. Our data support the hypothesis that LBP can inhibit cell responses to LPS by inhibiting LPS transfer from mCD14 to the Toll-like receptor 4-MD-2 signaling receptor. PMID- 12754216 TI - Localized cdc42 activation, detected using a novel assay, mediates microtubule organizing center positioning in endothelial cells in response to fluid shear stress. AB - Fluid flow regulates morphology, physiology, and pathophysiology of vascular endothelial cells (reviewed in Ref. 1). The small GTPase Cdc42 mediates polarity in several systems including migrating cells and early embryos, which involve reorientation of the microtubule organizing center (MTOC) and Golgi apparatus toward the direction of movement. Here, we show that Cdc42 is activated by fluid shear stress and that activation is a consequence of integrins binding to extracellular matrix. A novel fluorescence energy transfer assay to visualize Cdc42 activation in single cells shows that Cdc42 activity is polarized in the direction of flow. Localized activation of Cdc42 as well as the activity of Par6 and protein kinase Czeta direct the reorientation of the MTOC to a position on the downstream side of the nucleus relative to the direction of flow. Thus, shear stimulated integrin dynamics induce polarized Cdc42 activity, which induces MTOC localization through the Par6-protein kinase Czeta complex. PMID- 12754217 TI - Necrotic cell death in response to oxidant stress involves the activation of the apoptogenic caspase-8/bid pathway. AB - Human epithelial (A549) cells exposed to hyperoxia die by cellular necrosis. In the current study, we demonstrated the involvement of apoptogenic factors in epithelial cell necrosis in response to hyperoxia, including the formation of the Fas-related death-inducing signaling complex and initiation of mitochondria dependent apoptotic pathways. We showed increased activation of both Bid and Bax in A549 cells subjected to hyperoxia. Bax activation involved a Bid-assisted conformational change. We discovered that the response to hyperoxia in vivo predominantly involved the activation of the Bid/caspase-8 pathway without apparent increases in Bax expression. Disruption of the Bid pathway by gene deletion protected against cell death in vivo and in vitro. Likewise, inhibition of caspase-8 by Flip also protected against cell death. Taken together, we have demonstrated the involvement of apoptogenic factors in epithelial cell responses to hyperoxia, despite a final outcome of cellular necrosis. We have, for the first time, identified a predominant role for the caspase-8/Bid pathway in signaling associated with hyperoxic lung injury and cell death in vivo and in vitro. PMID- 12754218 TI - E2F modulates keratinocyte squamous differentiation: implications for E2F inhibition in squamous cell carcinoma. AB - E2F regulation is essential for normal cell cycle progression. Therefore, it is not surprising that squamous cell carcinoma cell lines (SCC) overexpress E2F1 and exhibit deregulated E2F activity when compared with normal keratinocytes. Indeed, deliberate E2F1 deregulation has been shown to induce hyperplasia and skin tumor formation. In this study, we report on a dual role for E2F as a mediator of keratinocyte proliferation and modulator of squamous differentiation. Overexpression of E2F isoforms in confluent primary keratinocyte cultures resulted in suppression of differentiation-associated markers. Moreover, we found that the DNA binding domain and the trans-activation domain of E2F1 are important in mediating suppression of differentiation. Use of a dominant/negative form of E2F1 (E2F d/n) found that E2F inhibition alone is sufficient to suppress the activity of proliferation-associated markers but is not capable of inducing differentiation markers. However, if the E2F d/n is expressed in differentiated keratinocytes, differentiation marker activity is further induced, suggesting that E2F may act as a modulator of squamous differentiation. We therefore examined the effects of E2F d/n in a differentiation-insensitive SCC cell line. We found that treatment with the differentiating agent, 12-O-tetradecanoyl phorbol-13-acetate (TPA), or expression of E2F d/n alone had no effect on differentiation markers. However, a combination of E2F d/n + TPA induced the expression of differentiation markers. Combined, these data indicate that E2F may play a key role in keratinocyte differentiation. These data also illustrate the unique potential of anti-E2F therapies in arresting proliferation and inducing differentiation of SCCs. PMID- 12754219 TI - Targeting membrane-localized focal adhesion kinase to focal adhesions: roles of tyrosine phosphorylation and SRC family kinases. AB - In the present study, we examined regulation of activated focal adhesion kinase localization in focal adhesions. By using focal adhesion kinase fused to an inert transmembrane anchor, we found that the focal contact targeting region within focal adhesion kinase was preserved in the membrane-targeted fusion protein. However, upon tyrosine phosphorylation, full-length focal adhesion kinase became excluded from focal adhesions. This negative regulation of localization could be abolished by mutating key amino acid residues of focal adhesion kinase shown previously to be involved in adhesion-mediated signal transduction. Hyper phosphorylation of endogenous focal adhesion kinase induced by pervanadate resulted in a similar reduction of localization at focal adhesions. We also show here that Src family kinases are essential for the phosphorylation-dependent exclusion of focal adhesion kinase from focal adhesions. We propose here a molecular model for the tyrosine phosphorylation-dependent regulation of focal adhesion kinase organization involving Src kinases and an inhibitory phosphorylation of the C-terminal (Tyr-925) tyrosine residue. PMID- 12754220 TI - Global gene expression profiling in Escherichia coli K12. The effects of oxygen availability and FNR. AB - The work presented here is a first step toward a long term goal of systems biology, the complete elucidation of the gene regulatory networks of a living organism. To this end, we have employed DNA microarray technology to identify genes involved in the regulatory networks that facilitate the transition of Escherichia coli cells from an aerobic to an anaerobic growth state. We also report the identification of a subset of these genes that are regulated by a global regulatory protein for anaerobic metabolism, FNR. Analysis of these data demonstrated that the expression of over one-third of the genes expressed during growth under aerobic conditions are altered when E. coli cells transition to an anaerobic growth state, and that the expression of 712 (49%) of these genes are either directly or indirectly modulated by FNR. The results presented here also suggest interactions between the FNR and the leucine-responsive regulatory protein (Lrp) regulatory networks. Because computational methods to analyze and interpret high dimensional DNA microarray data are still at an early stage, and because basic issues of data analysis are still being sorted out, much of the emphasis of this work is directed toward the development of methods to identify differentially expressed genes with a high level of confidence. In particular, we describe an approach for identifying gene expression patterns (clusters) obtained from multiple perturbation experiments based on a subset of genes that exhibit high probability for differential expression values. PMID- 12754221 TI - Various effects of paromomycin on tmRNA-directed trans-translation. AB - trans-Translation is an unusual translation in which tmRNA plays a dual function as a tRNA and an mRNA to relieve the stalled translation on the ribosome. In this study, we examined the effects of an aminoglycoside antibiotic, paromomycin, on several tmRNA-related events in vitro. The results of a chemical footprinting study indicated that paromomycin molecules bind tmRNA at G332/G333 in the tRNA domain and A316 in the middle of the long helix between tRNA and mRNA domains. Paromomycin bound at G332/G333 inhibited aminoacylation, and the inhibition was suppressed by the addition of SmpB, a tmRNA-binding protein. It was also found that paromomycin causes a shift of the translation resuming point on tmRNA by -1. The effect on initiation shift was canceled by a mutation at the paromomycin binding site in 16 S rRNA but not by mutations in tmRNA. A high concentration of paromomycin inhibited trans-translation, whereas it enhanced the initiation shifted trans-translation when SmpB was exogenously added or a mutation was introduced at 333. The effect of paromomycin on trans-translation differs substantially from that on canonical translation, in which it induces miscoding by modulating the A site of the decoding helix of the small subunit RNA of the ribosome. PMID- 12754222 TI - Inactivation of Mg chelatase during transition from anaerobic to aerobic growth in Rhodobacter capsulatus. AB - The facultative photosynthetic bacterium Rhodobacter capsulatus can adapt from an anaerobic photosynthetic mode of growth to aerobic heterotrophic metabolism. As this adaptation occurs, the cells must rapidly halt bacteriochlorophyll synthesis to prevent phototoxic tetrapyrroles from accumulating, while still allowing heme synthesis to continue. A likely control point is Mg chelatase, the enzyme that diverts protoporphyrin IX from heme biosynthesis toward the bacteriochlorophyll biosynthetic pathway by inserting Mg(2+) to form Mg-protoporphyrin IX. Mg chelatase is composed of three subunits that are encoded by the bchI, bchD, and bchH genes in R. capsulatus. We report that BchH is the rate-limiting component of Mg chelatase activity in cell extracts. BchH binds protoporphyrin IX, and BchH that has been expressed and purified from Escherichia coli is red in color due to the bound protoporphyrin IX. Recombinant BchH is rapidly inactivated by light in the presence of O(2), and the inactivation results in the formation of a covalent adduct between the protein and the bound protoporphyrin IX. When photosynthetically growing R. capsulatus cells are transferred to aerobic conditions, Mg chelatase is rapidly inactivated, and BchH is the component that is most rapidly inactivated in vivo when cells are exposed to aerobic conditions. The light- and O(2)-stimulated inactivation of BchH could account for the rapid inactivation of Mg chelatase in vivo and provide a mechanism for inhibiting the synthesis of bacteriochlorophyll during adaptation of photosynthetically grown cells to aerobic conditions while still allowing heme synthesis to occur for aerobic respiration. PMID- 12754223 TI - Functional subsets of the virB type IV transport complex proteins involved in the capacity of Agrobacterium tumefaciens to serve as a recipient in virB-mediated conjugal transfer of plasmid RSF1010. AB - The virB-encoded type IV transport complex of Agrobacterium tumefaciens mediates the transfer of DNA and proteins into plant cells, as well as the conjugal transfer of IncQ plasmids, such as RSF1010, between Agrobacterium strains. While several studies have indicated that there are physical interactions among the 11 VirB proteins, the functional significance of the interactions has been difficult to establish since all of the proteins are required for substrate transfer. Our previous studies, however, indicated that although all of the VirB proteins are required for the capacity of a strain to serve as an RSF1010 donor, only a subset of these proteins in the recipient is necessary to increase the conjugal frequency by 3 to 4 logs. The roles of particular groups of VirB proteins in this increased recipient activity were examined in the study reported here. Examination of the expression of subgroups of virB genes revealed that translation of virB6 is necessary for expression of downstream open reading frames. Expression of limited subsets of the VirB proteins in a recipient strain lacking the Ti plasmid revealed that the VirB7 to VirB10 proteins yield a subcomplex that is functional in the recipient assay but that the VirB1 to VirB4 proteins, as a group, dramatically increase this activity in strains expressing VirB7 to VirB10. Finally, the membrane distribution and cross-linking patterns of VirB10, but not of VirB8 or VirB9, in a strain expressing only VirB7 to VirB10 are significantly altered compared to the patterns of the wild type. These characteristics are, however, restored to the wild-type status by coexpression of VirB1 to VirB3. Taken together, these results define subsets of type IV transport complex proteins that are critical in allowing a strain to participate as a recipient in virB-mediated conjugal RSF1010 transfer. PMID- 12754224 TI - Phosphorylation of the lipid A region of meningococcal lipopolysaccharide: identification of a family of transferases that add phosphoethanolamine to lipopolysaccharide. AB - A gene, NMB1638, with homology to the recently characterized gene encoding a phosphoethanolamine transferase, lpt-3, has been identified from the Neisseria meningitidis genome sequence and was found to be present in all meningococcal strains examined. Homology comparison with other database sequences would suggest that NMB1638 and lpt-3 represent genes coding for members of a family of proteins of related function identified in a wide range of gram-negative species of bacteria. When grown and isolated under appropriate conditions, N. meningitidis elaborated lipopolysaccharide (LPS) containing a lipid A that was characteristically phosphorylated with multiple phosphate and phosphoethanolamine residues. In all meningococcal strains examined, each lipid A species contained the basal diphosphorylated species, wherein a phosphate group is attached to each glucosamine residue. Also elaborated within the population of LPS molecules are a variety of "phosphoforms" that contain either an additional phosphate residue, an additional phosphoethanolamine residue, additional phosphate and phosphoethanolamine residues, or an additional phosphate and two phosphoethanolamine residues in the lipid A. Mass spectroscopic analyses of LPS from three strains in which NMB1638 had been inactivated by a specific mutation indicated that there were no phosphoethanolamine residues included in the lipid A region of the LPS and that there was no further phosphorylation of lipid A beyond one additional phosphate species. We propose that NMB1638 encodes a phosphoethanolamine transferase specific for lipid A and propose naming the gene "lptA," for "LPS phosphoethenolamine transferase for lipid A." PMID- 12754225 TI - Transcription of bacteriophage PM2 involves phage-encoded regulators of heterologous origin. AB - Bacteriophage PM2 is the only described member of the Corticoviridae family. It is an icosahedral dsDNA virus with a membrane residing underneath the protein coat. PM2 infects some gram-negative Pseudoalteromonas spp. In the present study, we mapped the viral promoters and showed that the PM2 genome consists of three operons. Four new virus genes were assigned based on their function in transcription. Proteins P15 and P16 are shown to repress early transcription, and proteins P13 and P14 are shown to activate late transcription events. The early regulatory region, containing genes for proteins P15 and P16, as well as the newly identified early promoter region in PM2, has significant sequence similarity with the Pseudoalteromonas pAS28 plasmid. P14, the transcription activator for the structural genes, has a zinc finger motif homologous to archaeal and eukaryotic TFIIS-type regulatory factors. PMID- 12754226 TI - Contribution of aggregation-promoting factor to maintenance of cell shape in Lactobacillus gasseri 4B2. AB - Aggregation-promoting factor (APF) was originally described as a protein involved in the conjugation and autoaggregation of Lactobacillus gasseri 4B2, whose corresponding apf gene was cloned and sequenced. In this report, we identified and sequenced an additional apf gene located in the region upstream of the previously published one. Inactivation of both apf genes was unsuccessful, indicating that APF function may be essential for the cell. Overproduction of APF proteins caused drastic alteration in the cell shape of this strain. These cells were irregular, twisted, enlarged, and tightly bound in unbreakable clumps of chains. Down-regulation of APF synthesis was achieved by cloning of the apf2 promoter region on a high-copy-number plasmid, which recruited a putative apf activator. As a consequence, the shape of the corresponding recombinant cells was elongated (filamentous) and cell division sites were no longer visible. None of the induced changes in APF production levels was clearly correlated with modifications of the aggregation phenotype. This report shows, for the first time, that APF proteins are mainly critical for L. gasseri 4B2 cell shape maintenance. PMID- 12754227 TI - Impaired growth rates in milk of Lactobacillus helveticus peptidase mutants can be overcome by use of amino acid supplements. AB - To evaluate the contribution of intracellular peptidases to the growth of the 14 amino-acid (aa) auxotroph Lactobacillus helveticus CNRZ32, single- and multiple peptidase-deletion mutants were constructed. Two broad-specificity aminopeptidases (PepC and PepN) and X-prolyl dipeptidyl aminopeptidase (PepX) were inactivated through successive cycles of chromosomal gene replacement mutagenesis. The inactivation of all three peptidases in JLS247 ((Delta)pepC (Delta)pepN (Delta)pepX) did not affect the growth rate in amino acid-defined medium. However, the peptidase mutants generally had decreased specific growth rates when acquisition of amino acids required hydrolysis of the proteins in milk, the most significant result being a 73% increase in generation time for JLS247. The growth rate deficiencies in milk were overcome by amino acid supplements with some specificity to each of the peptidase mutants. For example, milk supplementation with Pro resulted in the most significant growth rate increase for (Delta)pepX strains and a 7-aa supplement (Asn, Cys, Ile, Pro, Ser, Thr, and Val) resulted in a JLS247 growth rate indistinguishable from that of the wild type. Our results show that characterization of the activities of the broad specificity aminopeptidases had little predictive value regarding the amino acid supplements found to enhance the milk growth rates of the peptidase mutant strains. These results represent the first determination of the physiological roles with respect to specific amino acid requirements for peptidase mutants grown in milk. PMID- 12754228 TI - How clonal is Staphylococcus aureus? AB - Staphylococcus aureus is an important human pathogen and represents a growing public health burden owing to the emergence and spread of antibiotic-resistant clones, particularly within the hospital environment. Despite this, basic questions about the evolution and population biology of the species, particularly with regard to the extent and impact of homologous recombination, remain unanswered. We address these issues through an analysis of sequence data obtained from the characterization by multilocus sequence typing (MLST) of 334 isolates of S. aureus, recovered from a well-defined population, over a limited time span. We find no significant differences in the distribution of multilocus genotypes between strains isolated from carriers and those from patients with invasive disease; there is, therefore, no evidence from MLST data, which index variation within the stable "core" genome, for the existence of hypervirulent clones of this pathogen. Examination of the sequence changes at MLST loci during clonal diversification shows that point mutations give rise to new alleles at least 15 fold more frequently than does recombination. This contrasts with the naturally transformable species Neisseria meningitidis and Streptococcus pneumoniae, in which alleles change between 5- and 10-fold more frequently by recombination than by mutation. However, phylogenetic analysis suggests that homologous recombination does contribute toward the evolution of this species over the long term. Finally, we note a striking excess of nonsynonymous substitutions in comparisons between isolates belonging to the same clonal complex compared to isolates belonging to different clonal complexes, suggesting that the removal of deleterious mutations by purifying selection may be relatively slow. PMID- 12754229 TI - Novel developmental genes, fruCD, of Myxococcus xanthus: involvement of a cell division protein in multicellular development. AB - Myxococcus xanthus is a gram-negative soil bacterium that undergoes multicellular development upon nutrient starvation. In the present study, two novel developmental genes, fruC and fruD, of M. xanthus were identified and characterized. The FruD protein has significant amino acid sequence similarity to the DivIVA proteins of many bacteria including Bacillus subtilis. Vegetative cells of the fruD mutant exhibited a filamentous phenotype. The fruC and fruD mutants displayed similar delayed-development phenotypes. The formation of tightly aggregated mounds by fruC and fruD mutants was slower than that by the wild-type strain. Spore formation by the fruC and fruD mutants initiated after 30 h poststarvation, whereas wild-type M. xanthus initiated spore formation after 18 h. The fruCD genes were constitutively expressed as an operon during vegetative growth and development. S1 mapping revealed that transcription initiation sites of the fruCD operon were located 114 (P1) and 55 bp (P2) upstream of the fruC initiation codon. Only the P1 promoter was active during vegetative growth, while both the P1 and P2 promoters were active during development. The FruD protein was produced as a cytoplasmic protein and formed an oligomer during vegetative growth and development. PMID- 12754230 TI - Genomic sequence of C1, the first streptococcal phage. AB - C(1), a lytic bacteriophage infecting group C streptococci, is one of the earliest-isolated phages, and the method of bacterial classification known as phage typing was defined by using this bacteriophage. We present for the first time a detailed analysis of this phage by use of electron microscopy, protein profiling, and complete nucleotide sequencing. This virus belongs to the Podoviridae family of phages, all of which are characterized by short, noncontractile tails. The C(1) genome consists of a linear double-stranded DNA molecule of 16,687 nucleotides with 143-bp inverted terminal repeats. We have assigned functions to 9 of 20 putative open reading frames based on experimental substantiation or bioinformatic analysis. Their products include DNA polymerase, holin, lysin, major capsid, head-tail connector, neck appendage, and major tail proteins. Additionally, we found one intron belonging to the HNH endonuclease family interrupting the apparent lysin gene, suggesting a potential splicing event yielding a functional lytic enzyme. Examination of the C(1) DNA polymerase suggests that this phage utilizes a protein-primed mechanism of replication, which is prominent in the phi29-like members of Podoviridae. Consistent with this evidence, we experimentally determined that terminal proteins are covalently attached to both 5' termini, despite the fact that no homology to known terminal proteins could be elucidated in any of our open reading frames. Likewise, comparative genomics revealed no close evolutionary matches, suggesting that the C(1) bacteriophage is a unique member of the Podoviridae. PMID- 12754231 TI - Distribution and organization of auxotrophic genes on the multichromosomal genome of Burkholderia multivorans ATCC 17616. AB - The Burkholderia multivorans strain ATCC 17616 carries three circular chromosomes with sizes of 3.4, 2.5, and 0.9 Mb. To determine the distribution and organization of the amino acid biosynthetic genes on the genome of this beta proteobacterium, various auxotrophic mutations were isolated using a Tn5 derivative that was convenient not only for the determination of its insertion site on the genome map but also for the structural analysis of the flanking regions. Analysis by pulsed-field gel electrophoresis revealed that 20 out of 23 insertion mutations were distributed on the 3.4-Mb chromosome. More detailed analysis of the his, trp, arg, and lys mutations and their flanking regions revealed the following properties of these auxotrophic genes: (i) all nine his genes were clustered on the 3.4-Mb chromosome; (ii) seven trp genes were organized within two distinct regions, i.e., a trpEGDC cluster on the 3.4-Mb chromosome and a trpFBA cluster on the 2.5-Mb chromosome; (iii) the leu gene cluster, leuCDB, was also located close to the trpFBA cluster; and (iv) lysA and argG genes were located on the 2.5-Mb chromosome, in contrast to the argH gene, which was located on the 3.4-Mb chromosome. Southern hybridization analysis, allelic exchange mutagenesis of ATCC 17616, and complementation tests demonstrated that all of the genes examined were functional and existed as a single copy within the genome. The present findings also indicated that the 2.5 Mb chromosome carried various auxotrophic genes with no structural or functional counterparts on the remaining two chromosomes. PMID- 12754232 TI - Concentration and assembly of the division ring proteins FtsZ, FtsA, and ZipA during the Escherichia coli cell cycle. AB - The concentration of the cell division proteins FtsZ, FtsA, and ZipA and their assembly into a division ring during the Escherichia coli B/r K cell cycle have been measured in synchronous cultures obtained by the membrane elution technique. Immunostaining of the three proteins revealed no organized structure in newly born cells. In a culture with a doubling time of 49 min, assembly of the Z ring started around minute 25 and was detected first as a two-dot structure that became a sharp band before cell constriction. FtsA and ZipA localized into a division ring following the same pattern and time course as FtsZ. The concentration (amount relative to total mass) of the three proteins remained constant during one complete cell cycle, showing that assembly of a division ring is not driven by changes in the concentration of these proteins. Maintenance of the Z ring during the process of septation is a dynamic energy-dependent event, as evidenced by its disappearance in cells treated with sodium azide. PMID- 12754233 TI - Genomic analysis and initial characterization of the chitinolytic system of Microbulbifer degradans strain 2-40. AB - The marine bacterium Microbulbifer degradans strain 2-40 produces at least 10 enzyme systems for degrading insoluble complex polysaccharides (ICP). The draft sequence of the 2-40 genome allowed a genome-wide analysis of the chitinolytic system of strain 2-40. The chitinolytic system includes three secreted chitin depolymerases (ChiA, ChiB, and ChiC), a secreted chitin-binding protein (CbpA), periplasmic chitooligosaccharide-modifying enzymes, putative sugar transporters, and a cluster of genes encoding cytoplasmic proteins involved in N-acetyl-D glucosamine (GlcNAc) metabolism. Each chitin depolymerase was detected in culture supernatants of chitin-grown strain 2-40 and was active against chitin and glycol chitin. The chitin depolymerases also had a specific pattern of activity toward the chitin analogs 4-methylumbelliferyl-beta-D-N,N'-diacetylchitobioside (MUF diNAG) and 4-methylumbelliferyl-beta-D-N,N',N"-triacetylchitotrioside (MUF triNAG). The depolymerases were modular in nature and contained glycosyl hydrolase family 18 domains, chitin-binding domains, and polycystic kidney disease domains. ChiA and ChiB each possessed polyserine linkers of up to 32 consecutive serine residues. In addition, ChiB and CbpA contained glutamic acid rich domains. At 1,271 amino acids, ChiB is the largest bacterial chitinase reported to date. A chitodextrinase (CdxA) with activity against chitooligosaccharides (degree of polymerization of 5 to 7) was identified. The activities of two apparent periplasmic (HexA and HexB) N-acetyl-beta-D glucosaminidases and one cytoplasmic (HexC) N-acetyl-beta-D-glucosaminidase were demonstrated. Genes involved in GlcNAc metabolism, similar to those of the Escherichia coli K-12 NAG utilization operon, were identified. NagA from strain 2 40, a GlcNAc deacetylase, was shown to complement a nagA mutation in E. coli K 12. Except for the GlcNAc utilization cluster, genes for all other components of the chitinolytic system were dispersed throughout the genome. Further examination of this system may provide additional insight into the mechanisms by which marine bacteria degrade chitin and provide a basis for future research on the ICP degrading systems of strain 2-40. PMID- 12754235 TI - Genes of Bacillus cereus and Bacillus anthracis encoding proteins of the exosporium. AB - The exosporium is the outermost layer of spores of Bacillus cereus and its close relatives Bacillus anthracis and Bacillus thuringiensis. For these pathogens, it represents the surface layer that makes initial contact with the host. To date, only the BclA glycoprotein has been described as a component of the exosporium; this paper defines 10 more tightly associated proteins from the exosporium of B. cereus ATCC 10876, identified by N-terminal sequencing of proteins from purified, washed exosporium. Likely coding sequences were identified from the incomplete genome sequence of B. anthracis or B. cereus ATCC 14579, and the precise corresponding sequence from B. cereus ATCC 10876 was defined by PCR and sequencing. Eight genes encode likely structural components (exsB, exsC, exsD, exsE, exsF, exsG, exsJ, and cotE). Several proteins of the exosporium are related to morphogenetic and outer spore coat proteins of B. subtilis, but most do not have homologues in B. subtilis. ExsE is processed from a larger precursor, and the CotE homologue appears to have been C-terminally truncated. ExsJ contains a domain of GXX collagen-like repeats, like the BclA exosporium protein of B. anthracis. Although most of the exosporium genes are scattered on the genome, bclA and exsF are clustered in a region flanking the rhamnose biosynthesis operon; rhamnose is part of the sugar moiety of spore glycoproteins. Two enzymes, alanine racemase and nucleoside hydrolase, are tightly adsorbed to the exosporium layer; they could metabolize small molecule germinants and may reduce the sensitivity of spores to these, limiting premature germination. PMID- 12754234 TI - The dithiol:disulfide oxidoreductases DsbA and DsbB of Rhodobacter capsulatus are not directly involved in cytochrome c biogenesis, but their inactivation restores the cytochrome c biogenesis defect of CcdA-null mutants. AB - The cytoplasmic membrane protein CcdA and its homologues in other species, such as DsbD of Escherichia coli, are thought to supply the reducing equivalents required for the biogenesis of c-type cytochromes that occurs in the periplasm of gram-negative bacteria. CcdA-null mutants of the facultative phototroph Rhodobacter capsulatus are unable to grow under photosynthetic conditions (Ps(-)) and do not produce any active cytochrome c oxidase (Nadi(-)) due to a pleiotropic cytochrome c deficiency. However, under photosynthetic or respiratory growth conditions, these mutants revert frequently to yield Ps(+) Nadi(+) colonies that produce c-type cytochromes despite the absence of CcdA. Complementation of a CcdA null mutant for the Ps(+) growth phenotype was attempted by using a genomic library constructed with chromosomal DNA from a revertant. No complementation was observed, but plasmids that rescued a CcdA-null mutant for photosynthetic growth by homologous recombination were recovered. Analysis of one such plasmid revealed that the rescue ability was mediated by open reading frame 3149, encoding the dithiol:disulfide oxidoreductase DsbA. DNA sequence data revealed that the dsbA allele on the rescuing plasmid contained a frameshift mutation expected to produce a truncated, nonfunctional DsbA. Indeed, a dsbA ccdA double mutant was shown to be Ps(+) Nadi(+), establishing that in R. capsulatus the inactivation of dsbA suppresses the c-type cytochrome deficiency due to the absence of ccdA. Next, the ability of the wild-type dsbA allele to suppress the Ps(+) growth phenotype of the dsbA ccdA double mutant was exploited to isolate dsbA independent ccdA revertants. Sequence analysis revealed that these revertants carried mutations in dsbB and that their Ps(+) phenotypes could be suppressed by the wild-type allele of dsbB. As with dsbA, a dsbB ccdA double mutant was also Ps(+) Nadi(+) and produced c-type cytochromes. Therefore, the absence of either DsbA or DsbB restores c-type cytochrome biogenesis in the absence of CcdA. Finally, it was also found that the DsbA-null and DsbB-null single mutants of R. capsulatus are Ps(+) and produce c-type cytochromes, unlike their E. coli counterparts, but are impaired for growth under respiratory conditions. This finding demonstrates that in R. capsulatus the dithiol:disulfide oxidoreductases DsbA and DsbB are not essential for cytochrome c biogenesis even though they are important for respiration under certain conditions. PMID- 12754236 TI - Sigma 54 levels and physiological control of the Pseudomonas putida Pu promoter. AB - The cellular levels of the alternative sigma factor sigma(54) of Pseudomonas putida have been examined in a variety of growth stages and culture conditions with a single-chain Fv antibody tailored for detection of scarce proteins. The levels of sigma(54) were also monitored in P. putida strains with knockout mutations in ptsO or ptsN, known to be required for the C-source control of the sigma(54)-dependent Pu promoter of the TOL plasmid. Our results show that approximately 80 +/- 26 molecules of sigma(54) exist per cell. Unlike that in relatives of Pseudomonas (e.g., Caulobacter), where fluctuations of sigma(54) determine adaptation and differentiation when cells face starvation, sigma(54) in P. putida remains unexpectedly constant at different growth stages, in nitrogen starvation and C-source repression conditions, and in the ptsO and ptsN mutant strains analyzed. The number of sigma(54) molecules per cell in P. putida is barely above the predicted number of sigma(54)-dependent promoters. These figures impose a framework on the mechanism by which Pu (and other sigma(54)-dependent systems) may become amenable to physiological control. PMID- 12754237 TI - Polar localization of replicon origins in the multipartite genomes of Agrobacterium tumefaciens and Sinorhizobium meliloti. AB - The origins of replication of many different bacteria have been shown to reside at specific subcellular locations, but the mechanisms underlying their positioning and segregation are still being elucidated. In particular, little is known about the replication of multipartite genomes in bacteria. We determined the cellular positions of the origins of the replicons in the alpha proteobacteria Agrobacterium tumefaciens and Sinorhizobium meliloti and found that they are located at the poles of the cells. Our work demonstrates the conserved extreme polar localization of circular chromosome origins in these alpha proteobacteria and is also the first to specify the cellular location of origin regions from the repABC family. The cellular location of a derivative of the RK2 plasmid is distinct from that of the alpha proteobacterium genomic replicon origins but is conserved across bacteria. Colocalization experiments with the genomic replicons of A. tumefaciens revealed that the repABC replicons, although preferentially positioned at the cell pole, colocalize only rarely. For the repABC replicons in this organism, occupying discrete spatial locations may contribute to their coexistence and stable inheritance. PMID- 12754238 TI - Modeling bacterial evolution with comparative-genome-based marker systems: application to Mycobacterium tuberculosis evolution and pathogenesis. AB - The comparative-genomic sequencing of two Mycobacterium tuberculosis strains enabled us to identify single nucleotide polymorphism (SNP) markers for studies of evolution, pathogenesis, and epidemiology in clinical M. tuberculosis. Phylogenetic analysis using these "comparative-genome markers" (CGMs) produced a highly unusual phylogeny with a complete absence of secondary branches. To investigate CGM-based phylogenies, we devised computer models to simulate sequence evolution and calculate new phylogenies based on an SNP format. We found that CGMs represent a distinct class of phylogenetic markers that depend critically on the genetic distances between compared "reference strains." Properly distanced reference strains generate CGMs that accurately depict evolutionary relationships, distorted only by branch collapse. Improperly distanced reference strains generate CGMs that distort and reroot outgroups. Applying this understanding to the CGM-based phylogeny of M. tuberculosis, we found evidence to suggest that this species is highly clonal without detectable lateral gene exchange. We noted indications of evolutionary bottlenecks, including one at the level of the PHRI "C" strain previously associated with particular virulence characteristics. Our evidence also suggests that loss of IS6110 to fewer than seven elements per genome is uncommon. Finally, we present population-based evidence that KasA, an important component of mycolic acid biosynthesis, develops G312S polymorphisms under selective pressure. PMID- 12754239 TI - Coaggregation-mediated interactions of streptococci and actinomyces detected in initial human dental plaque. AB - Streptococci and actinomyces that initiate colonization of the tooth surface frequently coaggregate with each other as well as with other oral bacteria. These observations have led to the hypothesis that interbacterial adhesion influences spatiotemporal development of plaque. To assess the role of such interactions in oral biofilm formation in vivo, antibodies directed against bacterial surface components that mediate coaggregation interactions were used as direct immunofluorescent probes in conjunction with laser confocal microscopy to determine the distribution and spatial arrangement of bacteria within intact human plaque formed on retrievable enamel chips. In intrageneric coaggregation, streptococci such as Streptococcus gordonii DL1 recognize receptor polysaccharides (RPS) borne on other streptococci such as Streptococcus oralis 34. To define potentially interactive subsets of streptococci in the developing plaque, an antibody against RPS (anti-RPS) was used together with an antibody against S. gordonii DL1 (anti-DL1). These antibodies reacted primarily with single cells in 4-h-old plaque and with mixed-species microcolonies in 8-h-old plaque. Anti-RPS-reactive bacteria frequently formed microcolonies with anti-DL1 reactive bacteria and with other bacteria distinguished by general nucleic acid stains. In intergeneric coaggregation between streptococci and actinomyces, type 2 fimbriae of actinomyces recognize RPS on the streptococci. Cells reactive with antibody against type 2 fimbriae of Actinomyces naeslundii T14V (anti-type-2) were much less frequent than either subset of streptococci. However, bacteria reactive with anti-type-2 were seen in intimate association with anti-RPS reactive cells. These results are the first direct demonstration of coaggregation mediated interactions during initial plaque accumulation in vivo. Further, these results demonstrate the spatiotemporal development and prevalence of mixed species communities in early dental plaque. PMID- 12754240 TI - Role of feedback regulation of pantothenate kinase (CoaA) in control of coenzyme A levels in Escherichia coli. AB - Pantothenate kinase (CoaA) is a key regulator of coenzyme A (CoA) biosynthesis in Escherichia coli, and its activity is controlled by feedback inhibition by CoA and its thioesters. The importance of feedback inhibition in the control of the intracellular CoA levels was tested by constructing three site-directed mutants of CoaA that were predicted to be feedback resistant based on the crystal structure of the CoaA-CoA binary complex. CoaA[R106A], CoaA[H177Q], and CoaA[F247V] were purified and shown to retain significant catalytic activity and be refractory to inhibition by CoA. CoaA[R106A] retained 50% of the catalytic activity of CoaA, whereas the CoaA[H177Q] and CoaA[F247V] mutants were less active. The importance of feedback control of CoaA to the intracellular CoA levels was assessed by expressing either CoaA or CoaA[R106A] in strain ANS3 [coaA15(Ts) panD2]. Cells expressing CoaA[R106A] had significantly higher levels of phosphorylated pantothenate-derived metabolites and CoA in vivo and excreted significantly more 4'-phosphopantetheine into the medium compared to cells expressing the wild-type protein. These data illustrate the key role of feedback regulation of pantothenate kinase in the control of intracellular CoA levels. PMID- 12754242 TI - Cysteine-scanning analysis of the dimerization domain of EnvZ, an osmosensing histidine kinase. AB - EnvZ and OmpR are a transmembrane sensor and its cognate response regulator, respectively, regulating the transcription of porin genes in response to medium osmolarity in Escherichia coli. The cytoplasmic domain of EnvZ (EnvZc) possesses both kinase and phosphatase activities and can be dissected into two functional domains, A and B. Here, we performed a cysteine-scanning analysis of domain A, a 67-residue central dimerization and phosphatase domain containing His-243 as the phosphorylation site, and we examined the effects of the cysteine substitution mutations on the enzymatic activities of domain A. The substitution mutations were made at 31 residues, from which 24 mutant domain A proteins were biochemically characterized. From the analysis of the phosphatase activity of purified mutant proteins, it was found that there are two regions in domain A which are important for this activity. Cysteine mutations in these regions dramatically reduce or completely abolish the phosphatase activity of domain A. The mutations that have the most-severe effects on domain A phosphatase activity also significantly reduce the phosphatase activity of EnvZc containing the same mutation. Using an in vitro complementation system with EnvZc(H243V), these cysteine mutants were further characterized for their autophosphorylation activities as well as their phosphotransfer activities. The results indicate that some mutations are specific either for the phosphatase activity or for the kinase activity. PMID- 12754241 TI - Type IV-like pili formed by the type II secreton: specificity, composition, bundling, polar localization, and surface presentation of peptides. AB - The secreton or type II secretion machinery of gram-negative bacteria includes several type IV pilin-like proteins (the pseudopilins) that are absolutely required for secretion. We previously reported the presence of a bundled pilus composed of the pseudopilin PulG on the surface of agar-grown Escherichia coli K 12 cells expressing the Klebsiella oxytoca pullulanase (Pul) secreton genes at high levels (N. Sauvonnet, G. Vignon, A. P. Pugsley, and P. Gounon, EMBO J. 19:2221-2228, 2000). We show here that PulG is the only pseudopilin in purified pili and that the phenomenon is not restricted to the Pul secreton reconstituted in E. coli or to PulG. For example, high-level expression of the endogenous E. coli gsp secreton genes caused production of bundled pili composed of the pseudopilin GspG, and the Pul secreton was able to form pili composed of PulG like proteins from secreton systems of other bacteria. PulG derivatives in which the C terminus was extended by the addition of eight different peptides were also assembled into pili and functioned in secretion. Three of the C-terminal peptides were shown to be exposed along the entire length of the assembled pili. Hence, the C terminus of PulG may represent a permissive site for the insertion of immunogenic epitopes or other peptide sequences. One of these PulG variants, with a six-histidine tag at its C terminus, formed nonpolar, nonbundled pili, suggesting that bundle formation and polar localization are not correlated with the ability of PulG to function in secretion. We propose that the PulG pilus is an artifactual manifestation of a periplasmic "pseudopilus" and that cycles of pseudopilus extension and retraction within the periplasm propel pullulanase through secretin channels in the outer membrane. Abnormally long pili that extend beyond the outer membrane are produced only when pilus length control and retraction are deregulated by overproduction of the major pseudopilus subunit (PulG). PMID- 12754243 TI - Open reading frame sso2387 from the archaeon Sulfolobus solfataricus encodes a polypeptide with protein-serine kinase activity. AB - The predicted polypeptide product of open reading frame sso2387 from the archaeon Sulfolobus solfataricus, SsoPK2, displayed several of the sequence features conserved among the members of the "eukaryotic" protein kinase superfamily. sso2387 was cloned, and its polypeptide product was expressed in Escherichia coli. The recombinant protein, rSsoPK2, was recovered in insoluble aggregates that could be dispersed by using high concentrations (5 M) of urea. The solubilized polypeptide displayed the ability to phosphorylate itself as well as several exogenous proteins, including mixed histones, casein, bovine serum albumin, and reduced carboxyamidomethylated and maleylated lysozyme, on serine residues. The source of this activity resided in that portion of the protein displaying homology to the catalytic domain of eukaryotic protein kinases. By use of mass spectrometry, the sites of autophosphorylation were found to be located in two areas, one immediately N terminal to the region corresponding to subdomain I of eukaryotic protein kinases, and the second N terminal to the presumed activation loop located between subdomains VII and VIII. Autophosphorylation of rSsoPK2 could be uncoupled from the phosphorylation of exogenous proteins by manipulation of the temperature or mutagenic alteration of the enzyme. Autophosphorylation was detected only at temperatures >or=60 degrees C, whereas phosphorylation of exogenous proteins was detectable at 37 degrees C. Similarly, replacement of one of the potential sites of autophosphorylation, Ser(548), with alanine blocked autophosphorylation but not phosphorylation of an exogenous protein, casein. PMID- 12754244 TI - Quantitation of mecA transcription in oxacillin-resistant Staphylococcus aureus clinical isolates. AB - The transcription of mecA, the gene required for oxacillin resistance in staphylococci, was quantified in a collection of 65 geographically and genetically diverse clinical and 8 defined laboratory Staphylococcus aureus isolates. mecA transcription was measured by real-time reverse transcription-PCR, confirmed by Northern blot analysis, and correlated with the presence and DNA sequence of the two mecA repressors, mecI and blaI. Isolates were first examined that contained mecI and/or blaI with wild-type sequence. BlaI provided significantly more repression of mecA transcription than did MecI, unrelated to blaI genetic location. Both together repressed mecA better than either one alone. In clinical isolates containing only wild-type mecI, mecA transcription repression was 10- to 25-fold less effective than that seen in previously studied constructs derived from strain N315. There was a difference in the mecI ribosomal binding site (RBS) between the clinical isolates (GGAA) and N315 (GGAG). The GGAA RBS was associated with 5.5- to 7.3-fold less mecA repression than GGAG in isogenic constructs. The values generated for wild-type repressors were compared to those in 26 isolates containing mecI mutations. mecA transcription appeared to be repressed only by BlaI in isolates with mecI nonsense and frameshift mutations. In contrast, mecI repression seemed to be partially or fully retained in many of the isolates with mecI and one isolate with blaI missense mutations, providing structure-function correlates with the site and type of mutation. We conclude that mecA repressor activity is highly variable in clinical S. aureus isolates due to mecI mutations, RBS polymorphisms, and unidentified genomic adaptations. PMID- 12754245 TI - Conserved eukaryotic histone-fold residues substituted into an archaeal histone increase DNA affinity but reduce complex flexibility. AB - Although the archaeal and eukaryotic nucleosome core histones evolved from a common ancestor, conserved lysine residues are present at DNA-binding locations in all four eukaryotic histones that are not present in the archaeal histones. Introduction of lysine residues at the corresponding locations into an archaeal histone, HMfB, generated a variant with increased affinity for DNA that formed more compact complexes with DNA. However, these complexes no longer facilitated the circularization of short DNA molecules and had lost the flexibility to wrap DNA alternatively in either a negative or positive supercoil. PMID- 12754246 TI - Tertiary structure of bacterial murein: the scaffold model. AB - Although the chemical structure and physical properties of peptidoglycan have been elucidated for some time, the precise three-dimensional organization of murein has remained elusive. Earlier published computer simulations of the bacterial murein architecture modeled peptidoglycan strands in either a regular (D. Pink, J. Moeller, B. Quinn, M. Jericho, and T. Beveridge, J. Bacteriol. 182: 5925-5930, 2000) or an irregular (A. Koch, J. Theor. Biol. 204: 533-541, 2000) parallel orientation with respect to the plasma membrane. However, after integrating published experimental data on glycan chain length distribution and the degree of peptide side chain cross-linking into this computer simulation, we now report that the proposed planar network of murein appears largely dysfunctional. In contrast, a scaffold model of murein architecture, which assumes that glycan strands extend perpendicularly to the plasma membrane, was found to accommodate published experimental evidence and yield a viable stress bearing matrix. Moreover, this model is in accordance with the well-established principle of murein assembly in vivo, i.e., sequential attachment of strands to the preexisting structure. For the first time, the phenomenon of division plane alternation in dividing bacteria can be reconciled with a computer model of the molecular architecture of murein. PMID- 12754247 TI - Error-prone polymerase, DNA polymerase IV, is responsible for transient hypermutation during adaptive mutation in Escherichia coli. AB - The frequencies of nonselected mutations among adaptive Lac(+) revertants of Escherichia coli strains with and without the error-prone DNA polymerase IV (Pol IV) were compared. This frequency was more than sevenfold lower in the Pol IV defective strain than in the wild-type strain. Thus, the mutations that occur during hypermutation are due to Pol IV. PMID- 12754248 TI - Genomic structure of the Salmonella enterica serovar Typhimurium DT 64 bacteriophage ST64T: evidence for modular genetic architecture. AB - The complete sequence of the double-stranded DNA genome of a serotype-converting temperate bacteriophage, ST64T, was determined. The 40,679-bp genomic sequence of ST64T has an overall GC content of 47.5% and was reminiscent of a number of lambdoid phages, in particular, P22. Inferred proteins of ST64T which exhibited a high degree of sequence similarity to P22 proteins (>90%) included the functional serotype conversion cassette, integrase, excisionase, Abc1, Abc2, early antitermination (gp24), NinD, NinH, NinZ, packaging (gp3 and gp2), head (with the exception of gp26, gp7, gp20, and gp16), and tail proteins. The putative immunity genes were highly related to those of Salmonella enterica serotype Typhimurium phage L, whereas the lysis genes were almost identical to those of S. enterica serovar Typhimurium PS3. PMID- 12754249 TI - The glycosyltransferase gene encoding the enzyme catalyzing the first step of mycothiol biosynthesis (mshA). AB - Mycothiol is the major thiol present in most actinomycetes and is produced from the pseudodisaccharide 1D-myo-inosityl 2-acetamido-2-deoxy-alpha-D glucopyranoside (GlcNAc-Ins). A transposon mutant of Mycobacterium smegmatis shown to be GlcNAc-Ins and mycothiol deficient was sequenced to identify a putative glycosyltransferase gene designated mshA. The ortholog in Mycobacterium tuberculosis, Rv0486, was used to complement the mutant phenotype. PMID- 12754250 TI - Synthesis and localization of the Salmonella SPI-1 type III secretion needle complex proteins PrgI and PrgJ. AB - An essential component of type III secretion systems (TTSS) is a supramolecular structure termed the needle complex. In Salmonella enterica, at least four proteins make up this structure: InvG, PrgH, PrgK, and PrgI. Another protein, PrgJ, is thought to play a role in the assembly of this structure, but its function is poorly understood. We have analyzed the expression and localization of PrgJ and the needle protein PrgI in different S. enterica serovar Typhimurium mutant strains. We found that the levels of PrgI and PrgJ were significantly reduced in a TTSS-deficient invA mutant strain and that the decreased levels were due to protein instability. In addition, we found that PrgJ, although associated with the needle complex in wild-type S. enterica serovar Typhimurium, was absent from needle complexes obtained from an invJ mutant strain, which exhibits very long needle substructures. We suggest that PrgJ is involved in capping the needle substructure of the needle complex. PMID- 12754251 TI - Cbl-mediated ubiquitinylation is required for lysosomal sorting of epidermal growth factor receptor but is dispensable for endocytosis. AB - Ligand-induced down-regulation controls the signaling potency of the epidermal growth factor receptor (EGFR/ErbB1). Overexpression studies have identified Cbl mediated ubiquitinylation of EGFR as a mechanism of ligand-induced EGFR down regulation. However, the role of endogenous Cbl in EGFR down-regulation and the precise step in the endocytic pathway regulated by Cbl remain unclear. Using Cbl /- mouse embryonic fibroblast cell lines, we demonstrate that endogenous Cbl is essential for ligand-induced ubiquitinylation and efficient degradation of EGFR. Further analyses using Chinese hamster ovary cells with a temperature-sensitive defect in ubiquitinylation confirm a crucial role of the ubiquitin machinery in Cbl-mediated EGFR degradation. However, internalization into early endosomes did not require Cbl function or an intact ubiquitin pathway. Confocal immunolocalization studies indicated that Cbl-dependent ubiquitinylation plays a critical role at the early endosome to late endosome/lysosome sorting step of EGFR down-regulation. These findings establish Cbl as the major endogenous ubiquitin ligase responsible for EGFR degradation, and show that the critical role of Cbl-mediated ubiquitinylation is at the level of endosomal sorting, rather than at the level of internalization. PMID- 12754252 TI - dSmurf selectively degrades decapentaplegic-activated MAD, and its overexpression disrupts imaginal disc development. AB - MAD plays an important role in decapentaplegic (DPP) signaling throughout Drosophila development. Despite a recent study describing the restriction of DPP signaling via putative ubiquitin E3 ligase dSmurf (1), the molecular mechanisms of how dSmurf affects DPP signaling remain unexplored. Toward this goal we demonstrated the degradation of phosphorylated MAD by dSmurf. dSmurf selectively interacted with MAD, but not Medea and Dad, and the MAD-dSmurf interaction was induced by constitutively active DPP type I receptor thickveins. Wild type dSmurf, but not its C1029A mutant, mediated ubiquitination-dependent degradation of MAD. Silencing of dSmurf using RNA interference stabilized MAD protein in Drosophila S2 cells. Targeted expression of dSmurf in various tissues abolished phosphorylated MAD and disrupted patterning and growth. In contrast, similar overexpression of inactive dSmurf(C1029A) showed no significant effects on development. We conclude that dSmurf specifically targets phosphorylated MAD to proteasome-dependent degradation and regulates DPP signaling during development. PMID- 12754253 TI - Transcriptional coactivator PRIP, the peroxisome proliferator-activated receptor gamma (PPARgamma)-interacting protein, is required for PPARgamma-mediated adipogenesis. AB - Nuclear receptor coactivator PRIP (peroxisome proliferators-activated receptor (PPARgamma)-interacting protein) appears to serve as a linker between cAMP response element-binding protein-binding protein (CBP/p300)anchored and PBP (PPARgamma-binding protein)-anchored coactivator complexes involved in the transcriptional activity of nuclear receptors. Disruption of PRIP and PBP genes results in embryonic lethality between embryonic day 11.5 and 12.5 (postcoitum), indicating that PRIP and PBP are essential and nonredundant coactivators. Both PRIP and PBP were initially identified as PPARgamma coactivators, suggesting a role for these molecules in PPARgamma-induced adipogenesis. PBP-/- mouse embryonic fibroblasts fail to exhibit PPARgamma-stimulated adipogenesis indicating that PBP is a downstream regulator of PPARgamma-mediated adipogenesis. We now show that PRIP-/- mouse embryonic fibroblasts are also refractory to PPARgamma-stimulated adipogenesis and fail to express adipogenic marker aP2, a PPARgamma-responsive gene. Chromatin immunoprecipitation assays reveal reduced association in PRIP-/- cells of PIMT (PRIP-binding protein) and PBP with aP2 gene promoter, suggesting that PRIP is required for the linking of CBP/p300-anchored cofactor complex with PBP-anchored mediator complex. These data indicate that PRIP, like PBP, is a downstream regulator of PPARgamma-mediated adipogenesis and that both these coactivators are required for the successful completion of adipogenic program. PMID- 12754254 TI - Sorcin regulates excitation-contraction coupling in the heart. AB - Sorcin is a penta-EF hand Ca2+-binding protein that associates with both cardiac ryanodine receptors and L-type Ca2+ channels and has been implicated in the regulation of intracellular Ca2+ cycling. To better define the function of sorcin, we characterized transgenic mice in which sorcin was overexpressed in the heart. Transgenic mice developed normally with no evidence of cardiac hypertrophy and no change in expression of other calcium regulatory proteins. In vivo hemodynamics revealed significant reductions in global indices of contraction and relaxation. Contractile abnormalities were also observed in isolated adult transgenic myocytes, along with significant depression of Ca2+ transient amplitudes. Whole cell ICa density and the time course of activation were normal in transgenic myocytes, but the rate of inactivation was significantly accelerated. These effects of sorcin on L-type Ca2+ currents were confirmed in Xenopus oocyte expression studies. Finally, we examined the expression of sorcin in normal and failing hearts from spontaneous hypertensive heart failure rats. In normal myocardium, sorcin extensively co-localized with ryanodine receptors at the Z-lines, whereas in myopathic hearts the degree of co-localization was markedly disrupted. Together, these data indicate that sorcin modulates intracellular Ca2+ cycling and Ca2+ influx pathways in the heart. PMID- 12754255 TI - The receptor tyrosine kinase Ror2 associates with the melanoma-associated antigen (MAGE) family protein Dlxin-1 and regulates its intracellular distribution. AB - The mammalian Ror family receptor tyrosine kinases, Ror1 and Ror2, play crucial roles in developmental morphogenesis. Although the functions of Ror1 and Ror2 are redundant, Ror2 exhibits more specific functions during development. We show that when expressed in mammalian cells, Ror2, but not Ror1, associates with the melanoma-associated antigen (MAGE) family protein, Dlxin-1, which is known to bind to the homeodomain proteins Msx2 and Dlx5 and regulate their transcriptional functions. This association requires the cytoplasmic C-terminal region of Ror2, containing proline-rich and serine/threonine-rich domains, and the C-terminal necdin homology domain of Dlxin-1. Interestingly, the cytoplasmic C-terminal region of Ror2 is missing in patients with brachydactyly type B. Interestingly, transient expression and immunohistochemical analyses reveal that both Dlxin-1 and Msx2 are co-localized in the nuclei in the absence of Ror2. In the presence of Ror2, Dlxin-1 is co-localized with Ror2 at the membranous compartments and Msx2 is retained in the nuclei. It was also found that the majority of cellular Dlxin-1 is retained in the membrane fractions of wild-type but not Ror2-/- mouse embryonic fibroblasts. Furthermore, we show that transcriptional activity of Msx2, irrespective of Ror2 kinase activity, is regulated by ectopic expression of Ror2 using a reporter plasmid containing the WIP element. Thus, Ror2 sequesters Dlxin-1 in membranous compartments, thereby affecting the transcriptional function of Msx2. PMID- 12754256 TI - Functional implications from an unexpected position of the 49-kDa subunit of NADH:ubiquinone oxidoreductase. AB - Membrane-bound complex I (NADH:ubiquinone oxidoreductase) of the respiratory chain is considered the main site of mitochondrial radical formation and plays a major role in many mitochondrial pathologies. Structural information is scarce for complex I, and its molecular mechanism is not known. Recently, the 49-kDa subunit has been identified as part of the "catalytic core" conferring ubiquinone reduction by complex I. We found that the position of the 49-kDa subunit is clearly separated from the membrane part of complex I, suggesting an indirect mechanism of proton translocation. This contradicts all hypothetical mechanisms discussed in the field that link proton translocation directly to redox events and suggests an indirect mechanism of proton pumping by redox-driven conformational energy transfer. PMID- 12754257 TI - Recruitment of sigma54-RNA polymerase to the Pu promoter of Pseudomonas putida through integration host factor-mediated positioning switch of alpha subunit carboxyl-terminal domain on an UP-like element. AB - The interactions between the sigma54-containing RNA polymerase (sigma54-RNAP) and the region of the Pseudomonas putida Pu promoter spanning from the enhancer to the binding site for the integration host factor (IHF) were analyzed both by DNase I and hydroxyl radical footprinting. A short Pu region centered at position -104 was found to be involved in the interaction with sigma54-RNAP, both in the absence and in the presence of IHF protein. Deletion or scrambling of the -104 region strongly reduced promoter affinity in vitro and promoter activity in vivo, respectively. The reduction in promoter affinity coincided with the loss of IHF mediated recruitment of the sigma54-RNAP in vitro. The experiments with oriented alpha sigma54-RNAP derivatives containing bound chemical nuclease revealed interchangeable positioning of only one of the two alpha subunit carboxyl terminal domains (alphaCTDs) both at the -104 region and in the surroundings of position -78. The addition of IHF resulted in perfect position symmetry of the two alphaCTDs. These results indicate that, in the absence of IHF, the sigma54 RNAP asymmetrically uses only one alphaCTD subunit to establish productive contacts with upstream sequences of the Pu promoter. In the presence of IHF induced curvature, the closer proximity of the upstream DNA to the body of the sigma54-RNAP can allow the other alphaCTD to be engaged in and thus favor closed complex formation. PMID- 12754258 TI - Certain metals trigger fibrillation of methionine-oxidized alpha-synuclein. AB - The aggregation and fibrillation of alpha-synuclein has been implicated as a key step in the etiology of Parkinson's disease and several other neurodegenerative disorders. In addition, oxidative stress and certain environmental factors, including metals, are believed to play an important role in Parkinson's disease. Previously, we have shown that methionine-oxidized human alpha-synuclein does not fibrillate and also inhibits fibrillation of unmodified alpha-synuclein (Uversky, V. N., Yamin, G., Souillac, P. O., Goers, J., Glaser, C. B., and Fink, A. L. (2002) FEBS Lett. 517, 239-244). Using dynamic light scattering, we show that the inhibition results from stabilization of the monomeric form of Met-oxidized alpha synuclein. We have now examined the effect of several metals on the structural properties of methionine-oxidized human alpha-synuclein and its propensity to fibrillate. The presence of metals induced partial folding of both oxidized and non-oxidized alpha-synucleins, which are intrinsically unstructured under conditions of neutral pH. Although the fibrillation of alpha-synuclein was completely inhibited by methionine oxidation, the presence of certain metals (Ti3+, Zn2+, Al3+, and Pb2+) overcame this inhibition. These findings indicate that a combination of oxidative stress and environmental metal pollution could play an important role in triggering the fibrillation of alpha-synuclein and thus possibly Parkinson's disease. PMID- 12754259 TI - A novel two-pore domain K+ channel, TRESK, is localized in the spinal cord. AB - To find a novel human ion channel gene we have executed an extensive search by using a human genome draft sequencing data base. Here we report a novel two-pore domain K+ channel, TRESK (TWIK-related spinal cord K+ channel). TRESK is coded by 385 amino acids and shows low homology (19%) with previously characterized two pore domain K+ channels. However, the most similar channel is TREK-2 (two-pore domain K+ channel), and TRESK also has two pore-forming domains and four transmembrane domains that are evolutionarily conserved in the two-pore domain K+ channel family. Moreover, we confirmed that TRESK is expressed in the spinal cord. Electrophysiological analysis demonstrated that TRESK induced outward rectification and functioned as a background K+ channel. Pharmacological analysis showed TRESK to be inhibited by previously reported K+ channel inhibitors Ba2+, propafenone, glyburide, lidocaine, quinine, quinidine, and triethanolamine. Functional analysis demonstrated TRESK to be inhibited by unsaturated free fatty acids such as arachidonic acid and docosahexaenoic acid. TRESK is also sensitive to extreme changes in extracellular and intracellular pH. These results indicate that TRESK is a novel two-pore domain K+ channel that may set the resting membrane potential of cells in the spinal cord. PMID- 12754260 TI - Probing electrostatic channeling in protozoal bifunctional thymidylate synthase dihydrofolate reductase using site-directed mutagenesis. AB - In this study we used site-directed mutagenesis to test the hypothesis that substrate channeling in the bifunctional thymidylate synthase-dihydrofolate reductase enzyme from Leishmania major occurs via electrostatic interactions between the negatively charged dihydrofolate produced at thymidylate synthase and a series of lysine and arginine residues on the surface of the protein. Accordingly, 12 charge reversal or charge neutralization mutants were made, with up to 6 putative channel residues changed at once. The mutants were assessed for impaired channeling using two criteria: a lag in product formation at dihydrofolate reductase and an increase in dihydrofolate accumulation. Surprisingly, none of the mutations produced changes consistent with impaired channeling, so our findings do not support the electrostatic channeling hypothesis. Burst experiments confirmed that the mutants also did not interfere with intermediate formation at thymidylate synthase. One mutant, K282E/R283E, was found to be thymidylate synthase-dead because of an impaired ability to form the covalent enzyme-methylene tetrahydrofolate-deoxyuridate complex prerequisite for chemical catalysis. PMID- 12754261 TI - Cap G, a gelsolin family protein modulating protective effects of unidirectional shear stress. AB - Atherosclerosis is a progressive and complex pathophysiological process occurring in large arteries. Although it is of multifactorial origin, the disease develops at preferential sites along the vasculature in regions experiencing specific hemodynamic conditions that are predisposed to endothelial dysfunction. The exact mechanisms allowing endothelial cells to discriminate between plaque-free and plaque-prone flows remain to be explored. To investigate such mechanisms, we performed a proteomic analysis on endothelial cells exposed in vitro to these two flow patterns. A few spots on the two-dimensional gel had an intensity that was differentially regulated by plaque-free versus plaque-prone flows. One of them was further investigated and identified as macrophage-capping protein (Cap G), a member of the gelsolin protein superfamily. A 2-fold increase of Cap G protein and a 5-fold increase of Cap G mRNA were observed in cells exposed to a plaque free flow as compared with static cultures. This increase was not observed in cells exposed to plaque-prone flow. Plaque-free flow induced a corresponding increase in nuclear and cytoskeletal-associated Cap G. Finally, overexpression of Cap G in transfection assays increased the motility potential of endothelial cells. These observations together with the known functions of Cap G suggest that Cap G may contribute to the protective effect exerted by plaque-free flow on endothelial cells. On the contrary, in cells exposed to a plaque-prone flow, no induction of Cap G expression could be observed. PMID- 12754262 TI - Storage and mobilization as antagonistic functional constraints on seed storage globulin evolution. AB - When seeds germinate nearly all the proteins are degraded in senescing storage tissue cells. All these proteins act as amino acid reserves which are mobilized to nourish the seedling. Nevertheless, the major amount of the seeds' protein reserve consists of a few enzymatically inactive, abundant, genuine storage proteins. In their metabolism the conflicting processes of biosynthesis, protein turnover and breakdown, are temporally separated. No degradation of correctly formed storage proteins was observed at the time of synthesis and accumulation during seed maturation. Breakdown takes place after a (long) period of rest when seeds germinate and seedlings start growing. At that time genuine storage proteins are no longer synthesized. Genuine storage proteins have evolved structural features permitting controlled temporal patterns of protection and proteolysis. The acquisition of inserted sequence stretches as sites accessible to limited proteolysis played a key role in the evolution of this control system and happened in coevolution of genuine storage proteins with specific proteinases. This can be deduced from the results of current research on the mechanisms of limited and unlimited proteolysis of storage globulins and on storage globulin evolution. The evolved system of controlled structure-function interplay between storage globulins and proteinases is part of a syndrome that, in addition, comprises differential compartmentation and gene expression of storage proteins and proteinases for controlling the total spatial and temporal patterns of globulin storage and mobilization in maturing and germinating seeds. PMID- 12754263 TI - Effect of glucose on assimilatory sulphate reduction in Arabidopsis thaliana roots. AB - With the aim of analysing the relative importance of sugar supply and nitrogen nutrition for the regulation of sulphate assimilation, the regulation of adenosine 5'-phosphosulphate reductase (APR), a key enzyme of sulphate reduction in plants, was studied. Glucose feeding experiments with Arabidopsis thaliana cultivated with and without a nitrogen source were performed. After a 38 h dark period, APR mRNA, protein, and enzymatic activity levels decreased dramatically in roots. The addition of 0.5% (w/v) glucose to the culture medium resulted in an increase of APR levels in roots (mRNA, protein and activity), comparable to those of plants kept under normal light conditions. Treatment of roots with d-sorbitol or d-mannitol did not increase APR activity, indicating that osmotic stress was not involved in APR regulation. The addition of O-acetyl-l-serine (OAS) also quickly and transiently increased APR levels (mRNA, protein, and activity). Feeding plants with a combination of glucose and OAS resulted in a more than additive induction of APR activity. Contrary to nitrate reductase, APR was also increased by glucose in N-deficient plants, indicating that this effect was independent of nitrate assimilation. [35S]-sulphate feeding experiments showed that the addition of glucose to dark-treated roots resulted in an increased incorporation of [35S] into thiols and proteins, which corresponded to the increased levels of APR activity. Under N-deficient conditions, glucose also increased thiol labelling, but did not increase the incorporation of label into proteins. These results demonstrate that (i) exogenously supplied glucose can replace the function of photoassimilates in roots; (ii) APR is subject to co ordinated metabolic control by carbon metabolism; (iii) positive sugar signalling overrides negative signalling from nitrate assimilation in APR regulation. Furthermore, signals originating from nitrogen and carbon metabolism regulate APR synergistically. PMID- 12754264 TI - Expression of the viviparous 1 (Pavp1) and p34cdc2 protein kinase (cdc2Pa) genes during somatic embryogenesis in Norway spruce (Picea abies [L.] Karst). AB - Detailed expression analysis of the Norway spruce (Picea abies [L.] Karst) Viviparous 1 (Pavp1) and p34cdc2 (cdc2Pa) genes was carried out during somatic embryogenesis. Pavp1, a gene associated with embryo development, was expressed in proliferating embryogenic suspension cultures in the absence of exogenous ABA. When somatic embryo formation was promoting by blocking proliferation, Pavp1 expression was reduced. During maturation, exogenous ABA induced increased Pavp1 expression, which peaked at the early cotyledonary stage of somatic embryogenesis. Following partial desiccation of mature somatic embryos at high relative humidity, Pavp1 expression persisted under germination conditions. Pavp1 expression was also detected in non-dormant immature male strobili and dormant terminal buds. These data confirm the functional conservation of Pavp1 during the evolution of seed plants and extend its function beyond the embryo. Cdc2Pa, a gene associated with the cell cycle, was up-regulated when the proliferation of embryogenic cells was blocked. Expression was again up-regulated in early embryogeny and again during germination. The implications of this up-regulation of cdc2Pa are discussed. PMID- 12754265 TI - A sucrose transporter, LjSUT4, is up-regulated during Lotus japonicus nodule development. AB - LjSUT4, encoding a putative sucrose transporter, was identified in a Lotus japonicus nodule cDNA library. The deduced amino acid sequence showed a high degree of identity with sucrose transporters from other plants. Semi-quantitative RT-PCR analysis demonstrated that the L. japonicus SUT4 gene was expressed at high levels in both roots and nodules. In situ hybridization revealed that, in young nodules, SUT4 mRNA transcripts are present in vascular bundles, inner cortex and both infected and uninfected cells while, in mature nodules, accumulation of transcripts was restricted only in vascular bundles and the inner cortex. The results indicated that LjSUT4 codes for a putative sucrose transporter, and its expression pattern suggests a possible shift in the mechanism of sugar transport during nodule development. The role of this polypeptide in sucrose transport and metabolism is discussed. PMID- 12754266 TI - Role of visible light in the recovery of photosystem II structure and function from ultraviolet-B stress in higher plants. AB - The effect of visible light on photosystem II reaction centre D1 protein in plants treated with ultraviolet-B light was studied. It was found that a 20 kDa C terminal fragment of D1 protein generated during irradiation with ultraviolet-B light was stable when plants were incubated in the dark, but was degraded when plants were incubated in visible light. In this condition the recovery of photosynthetic activity was also observed. Even a low level of white light was sufficient to promote both further degradation of the fragment and recovery of activity. During this phase, the D1 protein is the main synthesized thylakoid polypeptide, indicating that other photosystem II proteins are recycled in the recovery process. Although both degradation of the 20 kDa fragment and resynthesis of D1 are light-dependent phenomena, they are not closely related, as degradation of the 20 kDa fragment may occur even in the absence of D1 synthesis. Comparing chemical and physical factors affecting the formation of the fragment in ultraviolet-B light and its degradation in white light, it was concluded that the formation of the fragment in ultraviolet-B light is a photochemical process, whereas the degradation of the fragment in white light is a protease-mediated process. PMID- 12754267 TI - Arabidopsis 3-deoxy-D-manno-oct-2-ulosonate-8-phosphate synthase: cDNA cloning and expression analyses. AB - The molecular characterization of two isoforms of 3-deoxy-d-manno-oct-2-ulosonate (KDO) -8-phosphate synthase (AtkdsA1 and AtkdsA2) from Arabidopsis is reported here. First, by isolating a full-length cDNA for AtkdsA1, it was confirmed that the deduced primary structures of AtkdsA1 and AtkdsA2 proteins were 93% identical. Functional expression and purification studies demonstrated the efficient catalytic activity of the AtkdsA1 enzyme to produce KDO-8-phosphate from phosphoenolpyruvate and d-arabinose-5-phosphate. RT-PCR and RNA-gel blot analysis revealed different expression profiles for both genes; the AtkdsA1 gene was predominantly expressed in the shoots, while the AtkdsA2 transcript accumulated to a higher level in the roots, implicating differential roles of these isoforms in planta. PMID- 12754268 TI - Oxygen deficiency affects carbohydrate reserves in overwintering forage crops. AB - Anaerobic conditions developing under an ice cover affect winter survival and spring regrowth of economically important perennial crops. The objective was to compare, during a prolonged period of low (<2%) O2 at low temperature, the concentration of carbohydrates of four plant species contrasting in their resistance to oxygen deficiency. Four perennial forage species, lucerne (Medicago sativa L.), red clover (Trifolium pratense L.), timothy (Phleum pratense L.), and cocksfoot (Dactylis glomerata L.) were subjected to a progressively developing oxygen deficiency stress by enclosing potted plants in gas-tight bags in late autumn for overwintering in an unheated greenhouse. Timothy was previously reported to be more resistant to oxygen deficiency than the three other species. Non-structural carbohydrates increased and remained at a higher concentration in timothy than in the other three species under low O2 concentration. Concentrations of sucrose, fructose, glucose, and fructans increased in response to oxygen deficiency in timothy, whereas the concentration of soluble sugars decreased under the same conditions in lucerne, red clover, and cocksfoot. The gene expression of glyceraldehyde-3-phosphate dehydrogenase increased in response to low oxygen concentration in oxygen deficiency-sensitive lucerne while it remained unchanged in the oxygen deficiency-resistant timothy. It is concluded that timothy maintains higher carbohydrate reserves under oxygen deficiency, a specific feature that could favour its winter survival and spring regrowth. PMID- 12754270 TI - The RRASK motif in Xenopus cyclin B2 is required for the substrate recognition of Cdc25C by the cyclin B-Cdc2 complex. AB - The FLRRXSK sequence is conserved in the second cyclin box fold of B-type cyclins. We show that this conserved sequence in Xenopus cyclin B2, termed the RRASK motif, is required for the substrate recognition by the cyclin B-Cdc2 complex of Cdc25C. Mutations to charged residues of the RRASK motif of cyclin B2 abolished its ability to activate Cdc2 kinase without affecting its capacity to bind to Cdc2. Cdc2 bound to the cyclin B2 RRASK mutant was not dephosphorylated by Cdc25C, and as a result, the complex was inactive. The cyclin B2 RRASK mutants can form a complex with the constitutively active Cdc2, but a resulting active complex did not phosphorylate a preferred substrate Cdc25C in vitro, although it can phosphorylate the non-specific substrate histone H1. The RRASK mutations prevented the interaction of Cdc25C with the cyclin B2-Cdc2 complex. Consistently, the RRASK mutants neither induced germinal vesicle breakdown in Xenopus oocyte maturation nor activated in vivo Cdc2 kinase during the cell cycle in mitotic extracts. These results suggest that the RRASK motif in Xenopus cyclin B2 plays an important role in defining the substrate specificity of the cyclin B Cdc2 complex. PMID- 12754269 TI - Immunoglobulin light chains dictate vesicular transport-dependent and independent routes for IgM degradation by the ubiquitin-proteasome pathway. AB - Degradation of IgM mu heavy chains in light chain-negative pre-B cells is independent of vesicular transport, as is evident by its insensitivity to brefeldin A or cell permeabilization. Conversely, by the same criteria, degradation of the secretory mu heavy chain in light chain-expressing B cells depends on vesicular transport. To investigate whether the presence of conventional light chains or the developmental stage of the B-lymphocytes dictates the degradative route taken by mu, we express in 70Z/3 pre-B cells either lambda ectopically or kappa by lipopolysaccharides-stimulated differentiation into B cells and show their assembly with mu heavy chains. The resulting sensitivity of mu degradation to brefeldin A and cell permeabilization demonstrates that conventional light chains, a hallmark of B cell differentiation, are necessary and sufficient to divert mu from a vesicular transport-independent to a vesicular transport-dependent degradative route. Although both routes converge at the ubiquitin-proteasome degradation pathway, only in light chain-expressing cells is vesicular transport a prerequisite for mu ubiquitination. PMID- 12754271 TI - Amyloid beta-protein stimulates the expression of urokinase-type plasminogen activator (uPA) and its receptor (uPAR) in human cerebrovascular smooth muscle cells. AB - The accumulation of fibrillar amyloid-beta protein (A beta) in cerebral blood vessels, a condition known as cerebral amyloid angiopathy (CAA), is a key pathological feature of Alzheimer's disease and certain related disorders and is intimately associated with cerebrovascular cell death both in vivo and in vitro. Moreover, severe CAA leads to loss of vessel wall integrity and cerebral hemorrhage. Although the basis for these latter pathological consequences in CAA remains unresolved alterations in local proteolytic mechanisms may be involved. Here we show that pathogenic forms of A beta stimulate the expression of plasminogen activator activity in cultured human cerebrovascular smooth muscle (HCSM) cells, an in vitro model of CAA. RNase protection assays and plasminogen zymography showed that urokinase-type plasminogen activator (uPA) was responsible for this activity. There was preferential accumulation of uPA on the HCSM cell surface that was mediated through a concomitant increase in expression of the uPA receptor. In the presence of plasminogen there was robust degradation of A beta that was added to the HCSM cells resulting in restoration of cell viability. This suggests that increased expression of uPA may initially serve as a protective mechanism leading to localized degradation and clearance of the pathogenic stimulus A beta. On the other hand, chronic expression of uPA and plasminogen activation led to a profound loss of HCSM cell attachment. This suggests that a similar prolonged effect in vivo in the cerebral vessel wall may contribute to loss of integrity and cerebral hemorrhage in CAA. PMID- 12754272 TI - The chaperone environment at the cytoplasmic face of the endoplasmic reticulum can modulate rhodopsin processing and inclusion formation. AB - The human DnaJ (Hsp40) proteins HSJ1a and HSJ1b are type II DnaJ proteins with different C termini generated by alternate splicing. Both protein isoforms can regulate the ATPase activity and substrate binding of Hsp70. In this study, we have confirmed the neuronal expression of HSJ1a and HSJ1b proteins and localized their expression in human neural retina using isoform-specific antisera. HSJ1a and HSJ1b were enriched in photoreceptors, particularly the inner segments, but had different intracellular localization due to the prenylation of HSJ1b by a geranylgeranyl moiety. Because of their enrichment at the site of rhodopsin production, we investigated the effect of HSJ1 isoforms on the cellular processing of wild-type and mutant rhodopsin apoprotein in SK-N-SH cells. The expression of HSJ1b, but not HSJ1a, inhibited the normal cellular processing of wild-type rhodopsin-GFP, which co-localized with HSJ1b at the ER. HSJ1b expression also increased the incidence of inclusion formation by the wild-type protein. Both isoforms were recruited to mutant P23H rhodopsin inclusions, but only HSJ1b enhanced inclusion formation. Investigation of a prenylation-null mutant showed that the modulation of rhodopsin processing and inclusion formation was dependent on the correct subcellular targeting of HSJ1b to the cytosolic face of the ER. An Hsp70 interaction-null mutant of HSJ1b had the same effect as HSJ1b, suggesting that these phenomena were independent of Hsp70 and, furthermore, overexpression of Hsp70 with HSJ1b did not modulate the HSJ1b effect on inclusion formation, showing that Hsp70 was not limiting. The data provide evidence that cytoplasmic chaperones when targeted to the ER can influence the folding and processing of a GPCR and show that DnaJ protein function can be further specialized by alternative splicing and post-translational modification. PMID- 12754273 TI - A single intravenous dose of endotoxin rapidly alters serum lipoproteins and lipid transfer proteins in normal volunteers. AB - Endotoxemia is associated with rapid and marked declines in serum levels of LDL and HDL by unknown mechanisms. Six normal volunteers received a single, small intravenous (iv) dose of endotoxin (Escherichia coli 0113, 2 ng/kg) or saline in a random order, cross-over design. After endotoxin treatment, volunteers had mild, transient flu-like symptoms and markedly increased serum levels of tumor necrosis factor and its soluble receptors, interleukin-6, cortisol, serum amyloid A, and C-reactive protein. Triglyceride (TG), VLDL-TG, and nonesterified fatty acid increased (peak at 3-4 h), then TG declined (nadir at 9 h), and then cholesterol, LDL cholesterol, apolipoprotein B (apoB), and phospholipid declined (nadirs at 12-24 h). HDL cholesterol and apoA-I levels were not affected, but half of the decrease in phospholipid was HDL phospholipid. Lipopolysaccharide binding protein (LBP) rose 3-fold (peak at 12 h), with smaller and later decreases in the activities of phospholipid transfer protein and cholesteryl ester transfer protein. In conclusion, a decline in LDL was rapidly induced in normal volunteers with a single iv dose of endotoxin. The selective loss of phospholipid from HDL may have been mediated by LBP and, after more intense or prolonged inflammation, could result in increased HDL clearance and reduced HDL levels. PMID- 12754274 TI - Regulation of ABCA1 expression and cholesterol efflux during adipose differentiation of 3T3-L1 cells. AB - Adipose cells specialized in energy storage, contain large intracellular triglyceride-rich lipid droplets, are enriched with free cholesterol, and express sterol-regulated transcription factors such as liver X receptor (LXR). The recent identification of the LXR-dependent ATP binding cassette transporter A1 (ABCA1) pathway for cholesterol release from peripheral cells has led us to address the question of the expression and function of ABCA1 in adipocytes. In 3T3-L1 adipose cells, we observed a strong induction of ABCA1 mRNA during adipose differentiation, but only limited variations in ABCA1 protein. Lipid efflux onto apolipoprotein A-I (apoA-I), which depends on ABCA1, was comparable in adipocytes and preadipocytes, demonstrating a differential regulation of ABCA1 mRNA and cholesterol efflux. We also found that total cell cholesterol remained stable during differentiation of 3T3-L1 cells, but membrane cholesterol was lower in adipocytes than in preadipocytes, suggesting redistribution of cholesterol to the lipid droplet. Finally, we show that under standard lipolytic stimulation, 3T3-L1 adipocytes do not release cholesterol onto apoA-I, a process that required long exposures to lipolytic agents (24 h). In conclusion, despite large induction of ABCA1 mRNA during differentiation, cholesterol efflux through the ABCA1 pathway remains limited in adipocytes and requires prolonged lipolysis. This is consistent with the view of the adipocyte behaving as a cholesterol sink, with plasma cholesterol-buffering properties. PMID- 12754275 TI - Decreased PLTP mass but elevated PLTP activity linked to insulin resistance in HTG: effects of bezafibrate therapy. AB - Hypertriglyceridemia (HTG) is associated with insulin resistance, increased cholesteryl ester transfer (CET), and low HDL cholesterol. Phospholipid transfer protein (PLTP) may be involved in these relationships. Associations between CET, lipids, insulin resistance, CETP and PLTP activities, and PLTP mass were investigated in 18 HTG patients and 20 controls. Effects of 6 weeks of bezafibrate treatment were studied in HTG patients. HTG patients had higher serum triglycerides, insulin resistance, free fatty acid (FFA), and CET, lower levels of HDL cholesterol (-44%) and PLTP mass (-54%), and higher CETP (+20%) and PLTP activity (+48%) than controls. Bezafibrate reduced triglycerides, CET (-37%), insulin resistance (-53%), FFA (-48%), CETP activity (-12%), PLTP activity (-8%), and increased HDL cholesterol (+27%), whereas PLTP mass remained unchanged. Regression analysis showed a positive contribution of PLTP mass (P = 0.001) but not of PLTP activity to HDL cholesterol, whereas insulin resistance positively contributed to PLTP activity (P < 0.01). Bezafibrate-induced change in CET and HDL cholesterol correlated with changes in CETP activity and FFAs, but not with change in PLTP activity. Bezafibrate-induced change in PLTP activity correlated with change in FFAs (r = 0.455, P = 0.058). We propose that elevated PLTP activity in HTG is related to insulin resistance and not to increased PLTP mass. Bezafibrate-induced diminished insulin resistance is associated with a reduction of CET and PLTP activity. PMID- 12754276 TI - Lipophorin receptor-mediated lipoprotein endocytosis in insect fat body cells. AB - High-density lipophorin (HDLp) in the circulation of insects is able to selectively deliver lipids to target tissues in a nonendocytic manner. In Locusta migratoria, a member of the LDL receptor family has been identified and shown to mediate endocytosis of HDLp in mammalian cells transfected with the cDNA of this receptor. This insect lipophorin receptor (iLR) is temporally expressed in fat body tissue of young adult as well as larval locusts, as shown by Western blot analysis. Fluorescence microscopy revealed that fat body cells internalize fluorescently labeled HDLp and human receptor-associated protein only when iLR is expressed. Expression of iLR is down-regulated on Day 4 after an ecdysis. Consequently, HDLp is no longer internalized. By starving adult locusts immediately after ecdysis, we were able to prolong iLR expression. In addition, expression of the receptor was induced by starving adults after down-regulation of iLR. These results suggest that iLR mediates endocytosis of HDLp in fat body cells, and that expression of iLR is regulated by the demand of fat body tissue for lipids. PMID- 12754277 TI - Effect of a diet-induced n-3 PUFA depletion on cholinergic parameters in the rat hippocampus. AB - Because brain membranes contain large amounts of docosahexaenoic acid (DHA, 22:6n 3), and as (n-3) PUFA dietary deficiency can lead to impaired attention, learning, and memory performance in rodents, we have examined the influence of an (n-3) PUFA-deprived diet on the central cholinergic neurotransmission system. We have focused on several cholinergic neurochemical parameters in the frontal cortex and hippocampus of rats fed an (n-3) PUFA-deficient diet, compared with rats fed a control diet. The (n-3) PUFA deficiency resulted in changes in the membrane phospholipid compositions of both brain regions, with a dramatic loss (62-77%) of DHA. However, the cholinergic pathway was only modified in the hippocampus and not in the frontal cortex. The basal acetylcholine (ACh) release in the hippocampus of deficient rats was significantly (72%) higher than in controls, whereas the KCl-induced release was lower (34%). The (n-3) PUFA deprivation also caused a 10% reduction in muscarinic receptor binding. In contrast, acetylcholinesterase activity and the vesicular ACh transporter in both brain regions were unchanged. Thus, we evidenced that an (n-3) PUFA-deficient diet can affect cholinergic neurotransmission, probably via changes in the phospholipid PUFA composition. PMID- 12754278 TI - ApoE genotype-specific inhibition of apoptosis. AB - Endothelial cell apoptosis can be initiated by withdrawing growth factors or serum, and is inhibited by HDL. Our results show that the total lipoprotein population from apolipoprotein E 4/4 (APOE4/4) sera is less anti-apoptotic than total lipoproteins from other APOE genotypes, as measured by caspase 3/7 activity. Moreover, APOE4/4 VLDL antagonizes the antiapoptotic activity of HDL by a mechanism requiring binding of apoE4 on VLDL particles to an LDL family receptor. This ability of APOE4/4 VLDL to inhibit the antiapoptotic effects of HDL presents a potential mechanism by which the expression of several diseases, including atherosclerosis, is enhanced by the APOE4 genotype. PMID- 12754279 TI - Low-temperature effect on the sterol-dependent processing of SREBPs and transcription of related genes in HepG2 cells. AB - Lowering the growth temperature of HepG2 cells from 37 degrees C to 20 degrees C results in a 73% reduction in human squalene synthase (HSS) protein, a 76% reduction in HSS mRNA, and a 96% reduction in promoter activity of a secreted alkaline phosphatase-HSS reporter gene. A similar decrease in either mRNA or protein levels is observed for 3-hydroxy-3-methylglutaryl CoA reductase, farnesyl diphosphate synthase, the LDL receptor, and fatty acid synthase. All these proteins and mRNAs show either a decrease or a complete loss of sterol-dependent regulation in cells grown at 20 degrees C. In contrast, sterol regulatory element binding proteins (SREBPs)-1 and -2 exhibit a 2- to 3-fold increase in mRNA levels at 20 degrees C. The membrane-bound form of the SREBPs is dramatically increased, but the proteolytic processing to the nuclear (N-SREBP) form is inhibited under these conditions. Overexpression of the N-SREBP or SREBP cleavage-activating protein (SCAP), but not site-1 or site-2 proteases, restores the activation of the HSS promoter at 20 degrees C, most likely by liberating the SCAP-SREBP complex so that it can move to the Golgi for processing. These results indicate that the cholesterol synthesizing machinery is down-regulated at low temperatures, and points to the transport of the SCAP-SREBP complex to the Golgi as the specific down-regulated step. PMID- 12754281 TI - Nestin is expressed in vascular endothelial cells in the adult human pancreas. AB - In this study we examined the expression of nestin in islets, the exocrine part, and the big ducts of the adult human pancreas by immunofluorescent double staining. Two different anti-nestin antisera in combination with various pancreatic and endothelial markers were employed. Nestin-immunoreactive cells were found in islets and in the exocrine portion. All nestin-positive cells co expressed the vascular endothelial markers PECAM-1 (CD31), endoglin (CD105), and CD34 as well as vimentin. Endocrine, acinar, and duct cells did not stain for nestin. We also demonstrated that in the area of big pancreatic ducts, nestin positive cells represent small capillaries scattered in the connective tissue surrounding the duct epithelium and do not reside between the duct cells. We detected nestin-expressing endothelial cells located adjacent to the duct epithelium where endocrine differentiation occurs. We have shown that nestin is expressed by vascular endothelial cells in human pancreas, and therefore it is unlikely that nestin specifically marks a subpopulation of cells representing endocrine progenitors in the adult pancreas. PMID- 12754280 TI - Trans10, cis12-conjugated linoleic acid prevents triacylglycerol accumulation in adipocytes by acting as a PPARgamma modulator. AB - A group of polyunsaturated fatty acids called conjugated linoleic acids (CLAs) are found in ruminant products, where the most common isomers are cis9, trans11 (c 9,t11) and trans10, cis12 (t10,c12) CLA. A crude mixture of these isomers has been shown in animal studies to alter body composition by a reduction in body fat mass as well as an increase in lean body mass, with the t10,c12 isomer having the most pronounced effect. The objective of this study was to establish the molecular mechanisms by which t10,c12 CLA affects lipid accumulation in adipocytes. We have shown that t10,c12 CLA prevents lipid accumulation in human and mouse adipocytes at concentrations as low as 5 microM and 25 microM, respectively. t10,c12 CLA fails to activate peroxisome proliferator-activated receptor gamma (PPARgamma) but selectively inhibits thiazolidinedione-induced PPARgamma activation in 3T3-L1 adipocytes. Treatment of mature adipocytes with t10,c12 CLA alone or in combination with Darglitazone down-regulates the mRNA expression of PPARgamma as well as its target genes, fatty acid binding protein (aP2) and liver X receptor alpha (LXRalpha). Taken together, our results suggest that the trans10, cis12 CLA isomer prevents lipid accumulation in adipocytes by acting as a PPARgamma modulator. PMID- 12754282 TI - Ultrathin cryosections: an important tool for immunofluorescence and correlative microscopy. AB - Here we show that ultrathin cryosections of placental tissue can be used as a substrate in immunofluorescence experiments. A high degree of spatial resolution can be achieved in these preparations because there is essentially no out-of focus fluorescence. Therefore, immunofluorescence microscopy using ultrathin cryosections provides a very useful method for determining the precise subcellular localization of antigens in tissues. In addition, ultrathin cryosections of placenta also serve as a substrate for correlative immunofluorescence and immunoelectron microscopy using FluoroNanogold as the detection system. In correlative microscopy, the exact same structures in the same ultrathin section were observed by both fluorescence and electron microscopy. Using a particle counting procedure and electron microscopy, we compared the labeling obtained with colloidal gold and FluoroNanogold and found a higher number of particles with silver-enhanced FluoroNanogold than with colloidal gold. PMID- 12754283 TI - Enhanced serine palmitoyltransferase expression in proliferating fibroblasts, transformed cell lines, and human tumors. AB - Metastatic processes, including cell invasion, extracellular matrix degradation, and tissue remodeling, require cellular reorganization and proliferation. The cell signaling molecules required and the proteins involved in cell restructuring have not been completely elucidated. We have been studying the role of sphingolipids in normal cell activity and in several pathophysiological states. In this study we used immunohistochemistry to observe the presence of the two known subunits of serine palmitoyltransferase (SPT) in proliferating cells, in an in vitro model of wound repair, and in human malignant tissue. We report increased expression of the two subunits, SPT1 and SPT2, in the proliferating cells in these models. We also demonstrate a change in subcellular localization of the SPT subunits from predominantly cytosolic in quiescent cells to nuclear in proliferating cells. In addition, we observed SPT1 and SPT2 immunoreactivity in reactive stromal fibroblasts surrounding the carcinoma cells of some of the tumors. This enhanced SPT expression was absent in the stromal fibroblasts surrounding normal epithelial cells. Our results suggest a potential role for overexpression of SPT in the processes of cell metastasis. PMID- 12754284 TI - Differential expression of heparan sulfate domains in rat spleen. AB - The microarchitecture of the spleen is composed of a meshwork of reticulum cells and their matrix. Heparan sulfates (HS) are important components of this meshwork and are involved in processes such as cell adhesion, cell migration, and cytokine/growth factor binding. The expression of HS epitopes was analyzed using anti-HS antibodies. Four different staining patterns were observed, as exemplified by antibodies RB4EA12, HS4E4, AO4B08, and HS4C3. These antibodies recognize different chemical modifications in HS. In adult spleen, RB4EA12 stained only the reticular meshwork and blood vessels in the red pulp and marginal zone. HS4E4 stained blood vessel-associated basal lamina. AO4B08 and HS4C3 stained the reticular meshwork and blood vessels throughout the spleen, but only AO4B08 strongly stained smooth muscle cells and ring fibers. Interleukin-2 localized in the red pulp and marginal zone and was bound to HS. AO4B08, HS4C3, and RB4EA12 but not HS4E4 co-localized with interleukin-2. In 10-day-old spleen, HS4E4 recognized reticular fibers, which were not stained in the adult stage. Immunoelectron microscopy revealed that HS was restricted to basal laminae and reticular fibers. Taken together, data show that HS epitopes are differentially expressed in the spleen and that this may create specific extracellular environments for immunological processes. PMID- 12754285 TI - Bioluminescent detection of endotoxin effects on HIV-1 LTR-driven transcription in vivo. AB - We investigated the effects of Gram-negative bacterial lipopolysaccharide (LPS) on luciferase expression in transgenic reporter mice in which luciferase expression is driven by the nuclear factor kappaB (NF-kappaB)-dependent portion of the human immunodeficiency virus-1 (HIV-1) long terminal repeat (HIV-1 LTR). Using these mice, we dissected the sources of luciferase activity at the organ level by (a) assessing luciferase activity in organ homogenates, (b) bioluminescence imaging in vivo, and (c) bioluminescence imaging of individual organs ex vivo. Luciferin dosage was a critical determinant of the magnitude of photon emission from these reporter mice. Photon emission increased at doses from 0.5-6 mg of luciferin given by intraperitoneal (IP) injection. The differential between basal and LPS-induced bioluminescence was maximal at 3-6 mg of luciferin. Luciferase expression was highly inducible in lungs, liver, spleen, and kidneys after a single IP injection of LPS, as assessed by luciferase activity measurements in organ homogenates. Luciferase activity was also induced in the forebrain by treatment with IP LPS. In contrast, aerosolized LPS produced a response localized to the lungs as assessed by both bioluminescence and ex vivo luciferase assay measurements. These studies demonstrate the utility of luciferase reporter mice for determining organ-specific gene expression in response to local and systemic stimuli. PMID- 12754286 TI - Expression of catalase mRNA and protein in adult rat brain: detection by nonradioactive in situ hybridization with signal amplification by catalyzed reporter deposition (ISH-CARD) and immunohistochemistry (IHC)/immunofluorescence (IF). AB - Catalase, the classical peroxisomal marker enzyme, decomposes hydrogen peroxide and is involved in the antioxidant defense mechanisms of mammalian cells. In addition, catalase can oxidize, by means of its peroxidatic activity, a variety of substrates such as methanol and ethanol, producing the corresponding aldehydes. The involvement of brain catalase in the oxidation of ethanol is well established, and severe afflictions of the CNS in hereditary peroxisomal diseases (e.g., Zellweger syndrome) are well known. Whereas the distribution of catalase in the CNS has been investigated by enzyme histochemistry and immunohistochemistry (IHC), very little is known about the exact localization of catalase mRNA in brain. Here we report the application of a tyramine/CARD (catalyzed reporter deposition)-enhanced nonradioactive in situ hybridization (ISH) protocol for detection of catalase mRNA in sections of perfusion-fixed, paraffin-embedded rat brain. Catalase mRNA could be demonstrated in a large number of neurons throughout the rat brain as a distinct cytoplasmic staining signal with excellent morphological resolution. Compared to our standard ISH protocol, the CARD-enhanced protocol for catalase mRNA detection in rat brain showed higher sensitivity and significantly better signal-to-noise ratio. In parallel IHC experiments, using an antigen retrieval method consisting of combined trypsin digestion and microwave treatment of paraffin sections, the catalase antigen was found as distinct cytoplasmic granules in most catalase mRNA positive neurons. In addition, catalase-positive granules, presumably peroxisomes, were found by confocal laser scanning microscopy in glial cells, which were identified by double labeling immunofluorescence for GFAP and CNPase for astroglial cells and oligodentrocytes, respectively. The excellent preservation of morphology and sensitive detection of both mRNA and protein in our preparations warrant the application of the protocols described here for systematic studies of catalase and other peroxisomal proteins in diverse pathological conditions such as Alzheimer's disease and aging. PMID- 12754287 TI - Polypeptide GalNAc-transferases, ST6GalNAc-transferase I, and ST3Gal-transferase I expression in gastric carcinoma cell lines. AB - Mucin O-glycosylation in cancer is characterized by aberrant expression of immature carbohydrate structures leading to exposure of simple mucin-type carbohydrate antigens and peptide epitopes. Glycosyltransferases controlling the initial steps of mucin O-glycosylation are responsible for the altered glycosylation observed in cancer. We studied the expression in gastric cell lines of six UDP-GalNAc:polypeptide N-acetylgalactosaminyltransferases (GalNAc-T1, T2, T3, T4, T6, T11) that catalyze the initial key step in the regulation of mucin O glycosylation, the transfer of GalNAc from UDP-GalNAc to serine and threonine residues. We also studied the expression of ST6GalNAc-I, the enzyme responsible for the synthesis of Sialyl-Tn antigen (NeuAcalpha2,6GalNAc) and the ST3Gal-I, the enzyme responsible for the synthesis of Sialyl-T antigen (NeuAcalpha2,3Galbeta1,3GalNAc). This study was done using specific monoclonal antibodies, enzymatic assays, and RT-PCR. Our results showed that GalNAc-T1, -T2, and -T3 have an ubiquitous expression in all gastric cell lines, whereas GalNAc T4, -T6, and -T11 show a restricted expression pattern. The immunoreactivity with MAb VU-2-G7 suggests that, apart from GalNAc-T4, another GalNAc transferase is involved in the glycosylation of the Thr in the PDTR region of the MUC1 tandem repeat. The expression of ST3Gal-I correlates with the expression of the Sialyl-T antigen in gastric cell lines and in the control cell lines studied. The expression of ST6GalNAc-I is low in gastric cell lines, in accordance with the low/absent expression of the Sialyl-Tn antigen. PMID- 12754288 TI - Fixation methods for the study of lipid droplets by immunofluorescence microscopy. AB - The study of proteins associated with lipid droplets in adipocytes and many other cells is a rapidly developing area of inquiry. Although lipid droplets are easily visible by light microscopy, few standardized microscopy methods have been developed. Several methods of chemical fixation have recently been used to preserve cell structure before visualization of lipid droplets by light microscopy. We tested the most commonly used methods to compare the effects of the fixatives on cellular lipid content and lipid droplet structure. Cold methanol fixation has traditionally been used before visualization of cytoskeletal elements. We found this method unacceptable for study of lipid droplets because it extracted the majority of cellular phospholipids and promoted fusion of lipid droplets. Cold acetone fixation is similarly unacceptable because the total cellular lipids are extracted, causing collapse of the shell of lipid droplet-associated proteins. Fixation of cells with paraformaldehyde is the method of choice, because the cells retain their lipid content and lipid droplet structure is unaffected. As more lipid droplet-associated proteins are discovered and studied, it is critical to use appropriate methods to avoid studying artifacts. PMID- 12754289 TI - Patterns of MUC1 tissue expression defined by an anti-MUC1 cytoplasmic tail monoclonal antibody in breast cancer. AB - Our aim was to determine the pattern of expression of MUC1 mucin cytoplasmic tail (MUC1 CT) in breast carcinoma. A total of 98 invasive breast adenocarcinoma tumor samples were assayed by immunohistochemical (IHC) analysis. The pattern of reaction was classified as membrane, cytoplasmic, or mixed. Subcellular fractions were prepared after SDS-PAGE and Western blotting. The antibodies employed were anti-MUC1 CT (CT2 monoclonal antibody, MAb) and C595 MAb against the extracellular MUC1 core protein. With the CT2 MAb, IHC showed a high percentage of positive staining in 93% of specimens, with membrane staining the most common pattern observed. C595 MAb was reactive in 73% of specimens. Similar percentages of membrane and cytoplasmic staining were found, mainly in a mixed pattern. Western blotting showed different bands. With the CT2 MAb, the membrane fraction showed the most intense reaction; a strong band of reaction was detected at approximately <30 kD. With the C595 MAb, in most cases a double band at 200 kD was found. In breast epithelium, the pattern of MUC1 CT expression may constitute an indicator of MUC1 production because it does not depend on glycosylation. The pattern and extension of MUC1 CT positivity do not vary according to the histopathological subtype of the tumor. PMID- 12754290 TI - Skeletal muscles express the xenobiotic-metabolizing enzyme arylamine N acetyltransferase. AB - The human arylamine N-acetyltransferases (NATs) NAT1 and NAT2 are enzymes responsible for the acetylation of many arylamines and hydrazines, thereby playing an important role in both detoxification and activation of many drugs and carcinogens. Both enzymes show polymorphisms but exhibit key differences in substrate selectivity and tissue expression. In the present study, reverse transcriptase-PCR, Western blotting, and immunohistochemistry were used to investigate the expression of the NATs in human skeletal muscle. Despite the presence of its mRNA, NAT2 enzyme level was below the limit of detection. In contrast, both NAT1 mRNA and enzyme were readily detected in fetal, newborn, and adult muscles. In addition, punctate cytoplasmic and perinuclear NAT1 immunostaining was observed in all tissue sections, the staining being more intense in the fetal tissue. High expression of NAT1 enzyme in fetal muscle was also suggested by Western blotting. Because skeletal muscle accounts for a large proportion of body mass, muscle NAT1 expression may contribute significantly to the total activity in the body. These results further support the involvement of skeletal muscle in the metabolism of xenobiotics. PMID- 12754291 TI - Single-strand DNA aptamers as probes for protein localization in cells. AB - The accurate localization of proteins in fixed cells is important for many studies in cell biology, but good fixation is often antagonistic to good immunolabeling, given the density of well-preserved cells and the size of most labeled antibody probes. We therefore explored the use of single-stranded oligonucleotides (aptamers), which can bind to proteins with very high affinity and specificity but which are only approximately 10 kD. To evaluate these probes for general protein localization, we sought an aptamer that binds to a widely used protein tag, the green fluorescent protein (GFP). Although this quest was not successful, we were able to solve several practical problems that will confront any such labeling effort, e.g., the rates at which oligonucleotides enter fixed cells of different kinds and the extent of nonspecific oligonucleotide binding to both mammalian and yeast cell structures. Because such localization methods would be of particular value for electron microscopy of optimally fixed material, we also explored the solubility of aptamers under conditions suitable for freeze-substitution fixation. We found that aptamers are sufficiently soluble in cold organic solvents to encourage the view that this approach may be useful for the localization of specific proteins in context of cellular fine structure. PMID- 12754292 TI - Brain synaptic junctional proteins at the acrosome of rat testicular germ cells. AB - Proteins of the presynaptic exocytic machinery have been found associated with the acrosome of male germ cells, suggesting that the sperm acrosome reaction and neurotransmission at chemical synapses may share some common mechanisms. To substantiate this hypothesis, we studied the expression and ultrastructural localization of prominent pre- and postsynaptic protein components in rat testis. The presynaptic membrane trafficking proteins SV2 and complexin, the vesicular amino acid transporters VGLUT and VIAAT, the postsynaptic scaffolding protein ProSAP/Shank, and the postsynaptic calcium-sensor protein caldendrin, could be identified in germ line cells. Immunogold electron microscopy revealed an association of these proteins with the acrosome. In addition, evidence was obtained for the expression of the plasmalemmal glutamate transporters GLT1 and GLAST in rat sperm. The novel finding that not only presynaptic proteins, which are believed to be involved in membrane fusion processes, but also postsynaptic elements are present at the acrosome sheds new light on its structural organization. Moreover, our data point to a possible role for neuroactive amino acids in reproductive physiology. PMID- 12754293 TI - In situ localization of gelatinolytic activity in the extracellular matrix of metastases of colon cancer in rat liver using quenched fluorogenic DQ-gelatin. AB - Matrix metalloproteinases (MMPs) such as gelatinases are believed to play an important role in invasion and metastasis of cancer. In this study we investigated the possible role of MMP-2 and MMP-9 in an experimental model of colon cancer metastasis in rat liver. We demonstrated with gelatin zymography that the tumors contained MMP-2 and MMP-9, but only MMP-2 was present in the active form. Immunolocalization of MMP-2 showed that the protein was localized at basement membranes of colon cancer cells and in intratumor stroma, associated with extracellular matrix (ECM) components. However, zymography and immunohistochemistry (IHC) do not provide information on the localization of MMP activity. Therefore, we developed an in situ zymography technique using the quenched fluorogenic substrate DQ-gelatin in unfixed cryostat sections. The application of DQ-gelatin in combination with a gelled medium allows precise localization of gelatinolytic activity. Fluorescence due to gelatinolytic activity was found in the ECM of tumors and was localized similarly to both MMP-2 protein and collagen type IV, its natural substrate. The localization of MMP-2 activity and collagen type IV at similar sites suggests a role of MMP-2 in remodeling of ECM of stroma in colon cancer metastases in rat liver. PMID- 12754294 TI - Upregulation of cardiac insulin-like growth factor-I receptor by ACE inhibition after myocardial infarction: potential role in remodeling. AB - This study evaluated the effects of angiotensin-converting enzyme (ACE) inhibition after myocardial infarction (MI) on cardiac remodeling and gene expression and localization of components (ligands, receptors, and binding proteins) of the cardiac insulin-like growth factor (IGF) system. After ligation of the coronary artery, rats were randomized to vehicle or ACE inhibitor (Captopril, 50 mg/kg/day) for 4 weeks. Blood pressure, cardiac remodeling, and components of the IGF system were localized in the heart using in situ hybridization (ISH) and immunohistochemistry (IHC). The average infarct size was 42%. There were regional differences in the expression of the IGF system after MI, with increased IGF-I mRNA abundance in the border (24-fold) and infarct (12 fold) and increased IGF-binding protein (IGFBP)-3 mRNA in all areas of the failing left ventricle (threefold). Captopril reduced blood pressure, attenuated cardiac remodeling, and caused a threefold increase in IGF-I receptor mRNA and protein in infarct, border zone, and viable myocardium (p<0.01). Captopril had no effect on IGF-I mRNA but further increased IGFBP-3 mRNA and protein in the border zone, (p<0.05). The changes in the cardiac IGF system following MI are highly localized, persist for at least 4 weeks, and can be modulated by ACE inhibition. These data suggest that the benefits of ACE inhibitors in attenuation of cardiac remodeling may be mediated in part through increased expression of the IGF-I receptor sensitizing the myocardium to the positive effects of endogenous IGF-I. PMID- 12754295 TI - Stigmoid bodies contain type I receptor proteins SorLA/LR11 and sortilin: new perspectives on their function. AB - Stigmoid bodies (SBs) are structures in the cytoplasm of neurons. SBs are mostly found in the hypothalamic region of the rat and contain a protein called huntingtin-associated protein 1 (HAP1). In a recent publication, large cytoplasmic structures were shown to be immunoreactive for a type I receptor called SorLA/LR11. By light microscopic analysis, these structures appeared similar to SBs in size and in brain regional and subcellular localization. To determine whether these large puncta correspond to HAP1-containing SBs, we used antibodies specific to various domains of the apolipoprotein receptor LR11 to perform immunocytochemistry in rat and mouse brain tissue. Transfection studies using HeLa cells were conducted to demonstrate the specificity of the antibodies. We found that, in both species, antibodies to the domain II (or VSP10 for vacuolar sorting protein 10 domain) of LR11 immunoreact with large cytoplasmic structures. Co-localization immunolabeling experiments in rat brain tissue sections and in neuron cultures showed that these LR11-immunoreactive structures correspond to HAP1-positive SBs. Electron microscopy was performed in rat hypothalamus and further demonstrated the presence of LR11 in SBs and its co localization with HAP1. LR11-containing SBs were most abundant in the hypothalamus but were also found in many brainstem nuclei, thalamus, and hippocampus. Our data also show that sortilin, another transmembrane protein containing a VPS10 domain, localizes to large cytoplasmic puncta and is found in LR11-positive and Hap1-positive SBs in hypothalamic neuron cultures. PMID- 12754296 TI - Genome-wide analysis of organ-preferential metastasis of human small cell lung cancer in mice. AB - Although a number of molecules have been implicated in the process of cancer metastasis, the organ-selective nature of cancer cells is still poorly understood. To investigate this issue, we established a metastasis model in mice with multiple organ dissemination by i.v. injection of human small cell lung cancer (SBC-5) cells. We analyzed gene-expression profiles of 25 metastatic lesions from four organs (lung, liver, kidney, and bone) using a cDNA microarray representing 23,040 genes and extracted 435 genes that seemed to reflect the organ specificity of the metastatic cells and the cross-talk between cancer cells and microenvironment. Furthermore, we discovered 105 genes that might be involved in the incipient stage of secondary-tumor formation by comparing the gene expression profiles of metastatic lesions classified according to size (<1 or >2 mm) as either "micrometastases" or "macrometastases." This genome-wide analysis should contribute to a greater understanding of molecular aspects of the metastatic process in different microenvironments, and provide indicators for new strategies to predict and prevent metastasis. PMID- 12754297 TI - Overexpression of BAD potentiates sensitivity to tumor necrosis factor-related apoptosis-inducing ligand treatment in the prostatic carcinoma cell line LNCaP. AB - Here we show that LNCaP, which is resistant to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis, becomes sensitive to TRAIL after overexpression of full-length, wild-type BAD (BAD WT). TRAIL induces caspase-dependent cleavage of BAD WT that results in generation of a M(r) 15,000 protein. LNCaP stably expressing truncated BAD (tBAD) and cells expressing mutated BAD at the caspase cleavage site were less sensitive to TRAIL treatment when compared to LNCaP expressing BAD WT. Cytochrome c and Smac/DIABLO release from mitochondria into cytosol was found after TRAIL treatment only in cells overexpressing BAD WT. Furthermore, differences in phosphorylation of serine residues for BAD WT and tBAD were identified. BAD WT was phosphorylated at positions S136 and S155, whereas tBAD was phosphorylated at positions S112, S136, and S155. LNCaP stably expressing BAD mutated at serine 112 to alanine was less sensitive to TRAIL treatment when compared to LNCaP expressing BAD WT. Lastly, recombinant BAD cleaved by caspase-3 is a more potent inducer of cytochrome c and Smac/DIABLO release than BAD WT. In summary, BAD-mediated sensitivity of LNCaP to TRAIL depends on the phosphorylation status of BAD WT and tBAD. PMID- 12754298 TI - ARMER, apoptotic regulator in the membrane of the endoplasmic reticulum, a novel inhibitor of apoptosis. AB - We have identified a novel protein, apoptotic regulator in the membrane of the endoplasmic reticulum (ARMER), which protects HT1080 fibrosarcoma cells from apoptosis induced by various stimuli. We demonstrate that ARMER is an endoplasmic reticulum (ER) integral membrane protein with four predicted transmembrane domains and a COOH-terminal KKXX ER retrieval motif. We used an inducible expression system (pIND) to study the effects of regulated ARMER overexpression. Cells in which ARMER was overexpressed exhibited protection from multiple apoptotic inducers including serum starvation, doxorubicin, UV irradiation, tumor necrosis factor alpha, and the ER stressors brefeldin A, tunicamycin, and thapsigargin. Analysis of the caspase proteolytic cascade reveals that ARMER inhibits proteolysis of the caspase-9-specific fluorogenic substrate LEHD-AFC as well as endogenous substrates downstream of caspase-9; however, it does not inhibit cytochrome c release or cleavage of caspase-9 itself. Apoptotic stimuli cause endogenous levels of ARMER protein and RNA to decrease, leading to cell death; however, sustaining ARMER protein levels through exogenous expression inhibits apoptosis. These data suggest that ARMER is a novel ER integral membrane protein which protects cells by inhibiting caspase-9 activity and reveal a possible role for ARMER in cell survival. PMID- 12754299 TI - Participation of kin17 protein in replication factories and in other DNA transactions mediated by high molecular weight nuclear complexes. AB - The Homo sapiens kin17 ((HSA)kin17) protein is a chromatin-associated protein conserved during evolution and overproduced in certain human tumor cell lines. For the first time, immunoelectron microscopy analysis of endogenous (HSA)kin17 protein revealed an ultrastructural co-localization of (HSA)kin17 and bromodeoxyuridine (BrdUrd) at sites of DNA replication after either short (15 min) or long (120 min) pulses of BrdUrd labeling. After hydroxyurea (HU) or L mimosine (Mimo) block and withdrawal, we observed that (HSA)kin17 was recruited onto the chromatin during the re-entry and the progression in the S phase. These results are consistent with a major role of (HSA)kin17 protein in DNA replication factories. Other treatments hampering replication fork progression and/or inducing double-strand breaks also triggered an accumulation and a concentration of the chromatin-bound (HSA)kin17 protein into large intranuclear foci 24 h post treatment. Moreover, HU- and Mimo-induced (HSA)kin17 foci were retained in the nucleus after detergent extraction, suggesting a strong association with nuclear structures. Gel filtration analyses of cellular extracts showed that endogenous (HSA)kin17 protein co-eluted with both replication proteins RPA32 and RPA70 in a fraction containing complexes of M(r) 600,000. Interestingly, HU-induced G(1)-S arrest triggered an increase in the molecular weight of complexes containing (HSA)kin17 protein. Hence, treatments interfering with either initiation and/or elongation of DNA replication also recruited chromatin-bound (HSA)kin17 protein. We hypothesize that in the presence of unrepaired DNA damage, (HSA)kin17 protein concentrated into high molecular weight complexes probably to create a bridge that contributes to the harmonization of DNA replication and repair. PMID- 12754300 TI - Retinoic acid differentially regulates cancer cell proliferation via dose dependent modulation of the mitogen-activated protein kinase pathway. AB - The chemotherapeutic agent retinoic acid (RA) and its derivatives have been used to treat many tumor types. The antitumor effects of retinoids are in part due to their ability to inhibit proliferation of cancer cells. However, smokers receiving dietary vitamin A and beta carotene in chemoprevention studies had a higher incidence of lung cancer. These studies imply that lower doses of retinoids may have tumor-promoting activity. The effects of RA are mediated by a family of ligand-dependent transcription factors, the retinoic acid receptors (RARs) and the retinoid X receptors (RXR). We examined the effects of low- and high-dose RA treatment on proliferation of human squamous cell carcinoma lines in vitro. Low concentrations of RA (20 nM) increased proliferation of SCC lines by epidermal growth factor (EGF) activation of the mitogen-activated protein kinase ERK1. These changes were accompanied by increased expression of S- and G(2) phase cyclins and cyclin-dependent kinases (cdk), increased Rb phosphorylation, and increased E2F-1 DNA binding activity. In contrast, higher doses of RA (40 nM to 1 micro M) inhibited ERK1 expression, caused accumulation of G(1) phase cyclins and cdks, decreased Rb phosphorylation, and increased Rb/E2F-1 association. Overexpression of ERK1 or dominant negative ERK1 was sufficient to reproduce the effects of low- and high-dose RA, respectively. Treatment with receptor selective retinoids revealed that both RARalpha and RARgamma mediated the effects of RA on SCC lines. We concluded that low-dose RA induced proliferation by increased EGF signaling while higher concentrations inhibited cell division by decreasing ERK1 activation. PMID- 12754301 TI - Protein tyrosine phosphatase epsilon inhibits signaling by mitogen-activated protein kinases. AB - Mitogen-activated protein kinases (MAPKs) mediate signaling from the cell membrane to the nucleus following their phosphorylation at conserved threonine and tyrosine residues within their activation loops. We show that protein tyrosine phosphatase epsilon (PTP epsilon) inhibits ERK1 and ERK2 kinase activity and reduces their phosphorylation; in agreement, ERK phosphorylation is increased in fibroblasts and in mammary tumor cells from mice genetically lacking PTP epsilon. PTP epsilon inhibits events downstream of ERKs, such as transcriptional activation mediated by Elk1 or by the serum response element. PTP epsilon also inhibits transcriptional activation mediated by c-Jun and C/EBP binding protein (CHOP) but not that mediated by the unrelated NFkB, attesting that it is broadly active within the MAPK family but otherwise specific. The effect of PTP epsilon on ERKs is at least in part indirect because phosphorylation of the threonine residue in the ERK activation loop is reduced in the presence of PTP epsilon. Nonetheless, PTP epsilon is present in a molecular complex with ERK, providing PTP epsilon with opportunity to act on ERK proteins also directly. We conclude that PTP epsilon is a physiological inhibitor of ERK signaling. Slow induction of PTP epsilon and its lack of nuclear translocation following mitogenic stimulation suggest that PTP epsilon functions to prevent inappropriate activation and to terminate prolonged, rather than acute, activation of ERK in the cytosol. PMID- 12754302 TI - The role of phosphatidylinositol 3'-kinase and its downstream signals in erbB-2 mediated transformation. AB - We previously demonstrated that erbB-2-overexpressing human mammary epithelial (HME) cells exhibit several transformed phenotypes including growth factor independence, anchorage-independent growth, motility, and invasiveness. Because phosphatidylinositol 3'-kinase (PI3K) is a major target of erbB-2 activation, we tested the contribution that PI3K and its downstream signaling pathways make to these phenotypes. Utilizing a constitutively active form of PI3K, p110CAAX, we show that PI3K can mediate most phenotypes observed in erbB-2-overexpressing cells. To identify pathways leading from PI3K to specific phenotypes, we expressed constitutively active AKT or PTEN in erbB-2-overexpressing cells or in HME cells. HME cells expressing constitutively active AKT were growth factor independent, anchorage independent and motile, but not invasive. PTEN expression blocked erbB-2-mediated invasion but none of the other phenotypes. Rottlerin blocked invasion induced by p110CAAX and erbB-2, suggesting that protein kinase C delta (PKC-delta) is the downstream effector of PI3K responsible for the invasive capacity of the cells. Consistent with these observations, phospho-AKT remained detectable in erbB-2 cells treated with LY294002 or expressing exogenous PTEN, but was abolished by treatment with the p38MAP kinase inhibitor SB202190. Thus, both PI3K-dependent and p38MAP kinase-dependent pathways lead to activation of AKT, and activation of PKC-delta, via PI3K, mediates invasion. PMID- 12754303 TI - Profiling the global tyrosine phosphorylation state. AB - Protein tyrosine kinases and protein tyrosine phosphatases play a key role in cell signaling, and the recent success of specific tyrosine kinase inhibitors in cancer treatment strongly validates the clinical relevance of basic research on tyrosine phosphorylation. Functional profiling of the tyrosine phosphoproteome is likely to lead to the identification of novel targets for drug discovery and provide a basis for novel molecular diagnostic approaches. The ultimate aim of current mass spectrometry-based phosphoproteomic approaches is the comprehensive characterization of the phosphoproteome. However, current methods are not yet sensitive enough for routine detection of a large percentage of tyrosine phosphorylated proteins, which are generally of low abundance. In this article, we discuss alternative methods that exploit Src homology 2 (SH2) domains for profiling the tyrosine phosphoproteome. SH2 domains are small protein modules that bind specifically to tyrosine-phosphorylated peptides; there are more than 100 SH2 domains in the human genome, and different SH2 domains bind to different classes of tyrosine-phosphorylated ligands. These domains play a critical role in the propagation of signals in the cell, mediating the relocalization and complex formation of proteins in response to changes in tyrosine phosphorylation. We have developed an SH2 profiling method based on far-Western blotting, in which a battery of SH2 domains is used to probe the global state of tyrosine phosphorylation. Application to the classification of human malignancies suggests that this approach has potential as a molecular diagnostic tool. We also describe ongoing efforts to modify and improve SH2 profiling, including the development of a multiplexed assay system that will allow high-throughput functional profiling of the tyrosine phosphoproteome. PMID- 12754304 TI - New insights into the rat spermatogonial proteome: identification of 156 additional proteins. AB - Despite the essential role played by spermatogonia in testicular function, little is known about these cells. To improve our understanding of their biology, our group recently identified a set of 53 spermatogonial proteins using two dimensional (2-D) gel electrophoresis and mass spectrometry. To continue this work, we investigated a subset of the spermatogonial proteome using narrow range immobilized pH gradients to favor the detection of less abundant proteins. A 2-D reference map of spermatogonia in the pH range 4-9 was created, and protein entities fractionated in a pH 5-6 2-D gel were further processed for protein identification. A new set of 156 polypeptides was identified by peptide mass fingerprinting and tandem mass spectrometry. These polypeptides corresponded to 102 different proteins, which reflect the complexity of post-translational modifications. Seventy-nine of these proteins were identified for the first time in spermatogonia. All identified proteins were classified into functional groups. This work represents a first step toward the establishment of a systematic spermatogonia protein database. PMID- 12754305 TI - Efficient and specific removal of albumin from human serum samples. AB - Patient serum or plasma is frequently monitored for biochemical markers of disease or physiological status. Many of the rapidly evolving technologies of proteome analysis are being used to find additional clinically informative protein markers. The unusually high abundance of albumin in serum can interfere with the resolution and sensitivity of many proteome profiling techniques. We have used monoclonal antibodies against human serum albumin (HSA) to develop an immunoaffinity resin that is effective in the removal of both full-length HSA and many of the HSA fragments present in serum. This resin shows markedly better performance than dye-based resins in terms of both the efficiency and specificity of albumin removal. Immunoglobulins are another class of highly abundant serum protein. When protein G resin is used together with our immunoaffinity resin, Ig proteins and HSA can be removed in a single step. This strategy could be extended to the removal of any protein for which specific antibodies or affinity reagents are available. PMID- 12754306 TI - Exercise but not prostanoids enhance levels of vascular endothelial growth factor and other proliferative agents in human skeletal muscle interstitium. AB - In the present study we examined whether exercise and prostanoids have an effect on the muscle interstitial concentration of vascular endothelial growth factor (VEGF) and on the proliferative effect of muscle interstitial fluid. Dialysate from resting and exercising human skeletal muscle, obtained either during control conditions or during cyclooxygenase inhibition, was examined for its content of VEGF and for its effect on endothelial cell proliferation. Microdialysis probes with high (960 kDa) and low (5 kDa) molecular-mass cut-off membranes were placed in the vastus lateralis muscle of healthy young males. The subjects performed one legged knee extensions (20 W). The concentration of VEGF in the 960 kDa dialysate was greater (P < 0.05) during exercise compared to at rest (67 +/- 28 vs. 230 +/- 22 pg ml-1). The rate of endothelial cell proliferation was 2.7-fold higher (P < 0.05) with the 960 kDa dialysate from resting muscle than with perfusate and was 5.8-fold higher (P < 0.05) than the perfusate value with dialysate from exercising muscle. VEGF was not enhanced with exercise in the 5 kDa dialysate, yet the exercise dialysate induced a 1.9-fold higher (P < 0.05) proliferation than the resting dialysate. Cyclooxygenase inhibition did not affect the VEGF concentration or the proliferating effect of the dialysates (P > 0.05). This study demonstrates for the first time that VEGF is present in the interstitium of human skeletal muscle and that exercise enhances the interstitial concentration of VEGF and of other, as yet unidentified, angiogenic compounds. Products of cyclooxygenase do not appear to have an effect on the release of VEGF or other proliferative agents in human skeletal muscle. PMID- 12754308 TI - Interaction between sympathetic nerve activation and muscle fibre contraction in resistance vessels of hamster retractor muscle. AB - The interaction between skeletal muscle contraction and sympathetic nerve activation (SNA) on blood flow during exercise has remained ambiguous due to indirect estimates of vasomotor control. In the hamster retractor muscle (n=54), interactions between three levels of SNA (approximately 3, 6 and 12 Hz) and of contractile activity (2.5, 10 and 20 % duty cycle) were studied in feed arteries (FA) and first- (1A), second- (2A), and third-order (3A) arterioles using intravital microscopy. During functional dilatation with rhythmic muscle contractions, sympathetic vasoconstriction was sustained in FA and 1A but impaired in 2A and 3A (P<0.05), where vessels 'escaped' from responding to SNA. To account for changes in baseline diameter and blood flow during contractions, vasodilatation was induced passively (2-3 levels) in resting muscles with papaverine or sodium nitroprusside. Compared to functional dilatation, the range of passive dilatation was similar in 3A and progressively greater in 2A, 1A and FA. With passive dilatation, SNA responses were sustained in 2A and increased with baseline diameter in 3A. Blood flow through FA (rest, approximately 20 nl s( 1)) increased approximately 5-fold during contractile activity and approximately 10-fold during passive dilatation. Absolute flow reductions (nl s(-1)) with SNA increased during contractile activity and during passive dilatation; relative flow reductions were impaired during functional dilatation (P<0.05) and remained constant during passive dilatation. Thus, SNA can restrict blood flow to exercising muscle by constricting FA and 1A while dilatation prevails in 2A and 3A. Such concerted interaction will promote oxygen extraction when blood flow is restricted to maintain arterial pressure. PMID- 12754307 TI - Five ADNFLE mutations reduce the Ca2+ dependence of the mammalian alpha4beta2 acetylcholine response. AB - Five nicotinic acetylcholine receptor (nAChR) mutations are currently linked to autosomal dominant nocturnal frontal lobe epilepsy (ADNFLE). The similarity of their clinical symptoms suggests that a common functional anomaly of the mutations underlies ADNFLE seizures. To identify this anomaly, we constructed rat orthologues (S252F, +L264, S256L, V262L, V262M) of the human ADNFLE mutations, expressed them in Xenopus oocytes with the appropriate wild-type (WT) subunit (alpha4 or beta2), and studied the Ca2+ dependence of their ACh responses. All the mutations significantly reduced 2 mM Ca2+-induced increases in the 30 microM ACh response (P < 0.05). Consistent with a dominant mode of inheritance, this reduction persisted in oocytes injected with a 1:1 mixture of mutant and WT cRNA. BAPTA injections showed that the reduction was not due to a decrease in the secondary activation of Ca2+-activated Cl- currents. The S256L mutation also abolished 2 mM Ba2+ potentiation of the ACh response. The S256L, V262L and V262M mutations had complex effects on the ACh concentration-response relationship but all three mutations shifted the concentration-response relationship to the left at [ACh] >= 30 microM. Co-expression of the V262M mutation with a mutation (E180Q) that abolished Ca2+ potentiation resulted in 2 mM Ca2+ block, rather than potentiation, of the 30 microM ACh response, suggesting that the ADNFLE mutations reduce Ca2+ potentiation by enhancing Ca2+ block of the alpha4beta2 nAChR. Ca2+ modulation may prevent presynaptic alpha4beta2 nAChRs from overstimulating glutamate release at central excitatory synapses during bouts of synchronous, repetitive activity. Reducing the Ca2+ dependence of the ACh response could trigger seizures by increasing alpha4beta2-mediated glutamate release during such bouts. PMID- 12754309 TI - Facilitation of Ca(2+)-dependent exocytosis by Rac1-GTPase in bovine chromaffin cells. AB - Rho family GTPases are primary mediators of cytoskeletal reorganization, although they have also been reported to regulate cell secretion. Yet, the extent to which Rho family GTPases are activated by secretory stimuli in neural and neuroendocrine cells remains unknown. In bovine adrenal chromaffin cells, we found Rac1, but not Cdc42, to be rapidly and selectively activated by secretory stimuli using an assay selective for the activated GTPases. To examine effects of activated Rac1 on secretion, constitutively active mutants of Rac1 (Rac1-V12, Rac1-L61) were transiently expressed in adrenal chromaffin cells. These mutants facilitated secretory responses elicited from populations of intact and digitonin permeabilized cells as well as from cells under whole cell patch clamp. A dominant negative Rac1 mutant (Rac1-N17) produced no effect on secretion. Expression of RhoGDI, a negative regulator of Rac1, inhibited secretory responses while overexpression of effectors of Rac1, notably, p21-activated kinase (Pak1) and actin depolymerization factor (ADF) promoted evoked secretion. In addition, expression of effector domain mutants of Rac1-V12 that exhibit reduced activation of the cytoskeletal regulators Pak1 and Partner of Rac1 (POR1) resulted in a loss of Rac1-V12-mediated enhancement of evoked secretion. These findings suggest that Rac1, in part, functions to modulate secretion through actions on the cytoskeleton. Consistent with this hypothesis, the actin modifying drugs phalloidin and jasplakinolide enhanced secretion, while latrunculin-A inhibited secretion and eliminated the secretory effects of Rac1-V12. In summary, Rac1 was activated by secretory stimuli and modulated the secretory pathway downstream of Ca2+ influx, partly through regulation of cytoskeletal organization. PMID- 12754312 TI - Aquaporin-1 and HCO3(-)-Cl- transporter-mediated transport of CO2 across the human erythrocyte membrane. AB - Recent studies have suggested that aquaporin-1 (AQP1) as well as the HCO3(-)-Cl- transporter may be involved in CO2 transport across biological membranes, but the physiological importance of this route of gas transport remained unknown. We studied CO2 transport in human red blood cell ghosts at physiological temperatures (37 degrees C). Replacement of inert with CO2-containing gas above a stirred cell suspension caused an outside-to-inside directed CO2 gradient and generated a rapid biphasic intracellular acidification. The gradient of the acidifying gas was kept small to favour high affinity entry of CO2 passing the membrane. All rates of acidification except that of the approach to physicochemical equilibrium of the uncatalysed reaction were restricted to the intracellular environment. Inhibition of carbonic anhydrase (CA) demonstrated that CO2-induced acidification required the catalytic activity of CA. Blockade of the function of either AQP1 (by HgCl2 at 65 microM) or the HCO3(-)-Cl- transporter (by DIDS at 15 microM) completely prevented fast acidification. These data indicate that, at low chemical gradients for CO2, nearly the entire CO2 transport across the red cell membrane is mediated by AQP1 and the HCO3--Cl- transporter. Therefore, these proteins may function as high affinity sites for CO2 transport across the erythrocyte membrane. PMID- 12754311 TI - Sensory regulation of swallowing and airway protection: a role for the internal superior laryngeal nerve in humans. AB - During swallowing, the airway is protected from aspiration of ingested material by brief closure of the larynx and cessation of breathing. Mechanoreceptors innervated by the internal branch of the superior laryngeal nerve (ISLN) are activated by swallowing, and connect to central neurones that generate swallowing, laryngeal closure and respiratory rhythm. This study was designed to evaluate the hypothesis that the ISLN afferent signal is necessary for normal deglutition and airway protection in humans. In 21 healthy adults, we recorded submental electromyograms, videofluoroscopic images of the upper airway, oronasal airflow and respiratory inductance plethysmography. In six subjects we also recorded pressures in the hypopharynx and upper oesophagus. We analysed swallows that followed a brief infusion (4-5 ml) of liquid barium onto the tongue, or a sip (1-18 ml) from a cup. In 16 subjects, the ISLN was anaesthetised by transcutaneous injection of bupivacaine into the paraglottic compartment. Saline injections using the identical procedure were performed in six subjects. Endoscopy was used to evaluate upper airway anatomy, to confirm ISLN anaesthesia, and to visualise vocal cord movement and laryngeal closure. Comparisons of swallowing and breathing were made within subjects (anaesthetic or saline injection vs. control, i.e. no injection) and between subjects (anaesthetic injection vs. saline injection). In the non-anaesthetised condition (saline injection, 174 swallows in six subjects; no injection, 522 swallows in 20 subjects), laryngeal penetration during swallowing was rare (1.4 %) and tracheal aspiration was never observed. During ISLN anaesthesia (16 subjects, 396 swallows), all subjects experienced effortful swallowing and an illusory globus sensation in the throat, and 15 subjects exhibited penetration of fluid into the larynx during swallowing. The incidence of laryngeal penetration in the anaesthetised condition was 43 % (P < 0.01, compared with either saline or no injection) and of these penetrations, 56 % led to tracheal aspiration (without adverse effects). We further analysed the swallow cycle to evaluate the mechanism(s) by which fluid entered the larynx. Laryngeal penetration was not caused by premature spillage of oral fluid into the hypopharynx, delayed clearance of fluid from the hypopharynx, or excessive hypopharyngeal pressure generated by swallowing. Furthermore, there was no impairment in the ability of swallowing to halt respiratory airflow during the period of pharyngeal bolus flow. Rather, our observations suggest that loss of airway protection was due to incomplete closure of the larynx during the pharyngeal phase of swallowing. In contrast to the insufficient closure during swallowing, laryngeal closure was robust during voluntary challenges with the Valsalva, Muller and cough manoeuvres under ISLN anaesthesia. We suggest that an afferent signal arising from the ISLN receptor field is necessary for normal deglutition, especially for providing feedback to central neural circuits that facilitate laryngeal closure during swallowing. The ISLN afferent signal is not essential for initiating and sequencing the swallow cycle, for co-ordinating swallowing with breathing, or for closing the larynx during voluntary manoeuvres. PMID- 12754315 TI - Activation of presynaptic glycine receptors facilitates glycine release from presynaptic terminals synapsing onto rat spinal sacral dorsal commissural nucleus neurons. AB - Glycine is a major inhibitory neurotransmitter in the spinal cord and brainstem. Here we report the novel finding that presynaptic glycine autoreceptors modulate release from terminals synapsing onto rat spinal sacral dorsal commissural nucleus (SDCN) neurons. In mechanically dissociated SDCN neurons, in which functional presynaptic nerve terminals remain adherent to the isolated neurons, exogenously applied glycine (3 microM) increased the frequency of glycinergic spontaneous inhibitory postsynaptic currents (sIPSCs) without affecting their amplitudes or decay times. This suggests that glycine acts presynaptically to increase glycine release probability. Picrotoxin, at a concentration that had little direct effect on sIPSC frequency and amplitude (30 microM), significantly attenuated glycine-induced presynaptic sIPSC facilitation. The glycine-induced sIPSC frequency facilitation was completely abolished either in a Ca(2+)-free external solution or in the presence of 100 microM Cd2+, suggesting the involvement of extracellular Ca2+ influx into the nerve terminals. The glycine action was also completely occluded in the presence of 300 nM tetrodotoxin. In recordings from SDCN neurons in spinal cord slices, glycine (10 microM) increased evoked IPSC (eIPSC) amplitude and decreased the extent of paired-pulse facilitation. In response to brief high frequency stimulus trains the eIPSCs displayed a profound frequency-dependent facilitation that was greatly reduced by picrotoxin (30 microM). These results indicate that glycine acts at presynaptic autoreceptors, causing depolarization of the glycinergic nerve terminals, the subsequent activation of voltage-dependent Na+ and Ca2+ channels, and facilitation of glycine release. Furthermore, this presynaptic facilitation was observed under more physiological conditions, suggesting that these glycinergic autoreceptors may contribute to the integration of local inhibitory inputs to SDCN neurons. PMID- 12754316 TI - Temperature dependence of NADPH oxidase in human eosinophils. AB - The phagocyte NADPH oxidase helps kill pathogens by producing superoxide anion, O2-. This enzyme is electrogenic because it translocates electrons across the membrane, generating an electron current, Ie. Using the permeabilized patch voltage-clamp technique, we studied the temperature dependence of Ie in human eosinophils stimulated by phorbol myristate acetate (PMA) from room temperature to >37 degrees C. For comparison, NADPH oxidase activity was assessed by cytochrome c reduction. The intrinsic temperature dependence of the assembled, functioning NADPH oxidase complex measured during rapid temperature increases to 37 degrees C was surprisingly weak: the Arrhenius activation energy Ea was only 14 kcal mol(-1) (Q10, 2.2). In contrast, steady-state NADPH oxidase activity was strongly temperature dependent at 20-30 degrees C, with Ea 25.1 kcal mol(-1) (Q10, 4.2). The maximum Ie measured at 34 degrees C was -30.5 pA. Above 30 degrees C, the temperature dependence of both Ie and O2- production was less pronounced. Above 37 degrees C, Ie was inhibited reversibly. After rapid temperature increases, a secondary increase in Ie ensued, suggesting that high temperature promotes assembly of additional NADPH oxidase complexes. Evidently, about twice as many NADPH oxidase complexes are active near 37 degrees C than at 20 degrees C. Thus, the higher Q10 of steady-state Ie reflects both increased activity of each NADPH oxidase complex and preferential assembly of NADPH oxidase complexes at high temperature. In summary, NADPH oxidase activity in intact human eosinophils is maximal precisely at 37 degrees C. PMID- 12754318 TI - Morphine and endomorphins differentially regulate micro-opioid receptor mRNA in SHSY-5Y human neuroblastoma cells. AB - A sensitive quantitative-competitive reverse transcriptase-polymerase chain reaction method was developed to measure micro-opioid receptor (MOR) mRNA expression in SHSY-5Y neuroblastoma cells. Differentiation of SHSY-5Y cells with either retinoic acid (RA) or 12-o-tetradecanoyl-phorbol-13-acetate (TPA) significantly increased MOR mRNA levels. Morphine treatment (10 microM) for 24 h decreased MOR mRNA levels in control, as well as RA- and TPA-differentiated cells. In contrast, chronic exposure to the opioid peptides endomorphin-1 or endomorphin-2 significantly increased MOR mRNA levels in undifferentiated and RA differentiated cells. An opioid antagonist, naloxone, reversed the morphine and endomorphin-1 and -2 effects on MOR mRNA levels in undifferentiated SHSY-5Y cells, but naloxone had differential reversing effects on the agonists' regulation of MOR mRNA in RA- or TPA-differentiated cells. To investigate whether the changes in MOR mRNA expression paralleled changes in MOR receptor function, intracellular cAMP accumulation in SHSY-5Y cells was measured. After chronic treatment with morphine, forskolin-induced cAMP levels in SHSY-5Y cells were significantly higher than those of untreated control cells. In contrast, forskolin-induced cAMP accumulation levels were lower in cells treated with endomorphin-1 or -2 than in untreated control cells. Together, our studies indicate that the opioid alkaloid morphine and the opioid peptides endomorphin-1 and -2 differentially regulate MOR mRNA expression and MOR function in SHSY-5Y cells. PMID- 12754322 TI - Clinical evaluation and investigation of neuropathy. PMID- 12754319 TI - Effects of reboxetine on sympathetic neuroeffector transmission in rabbit carotid artery. AB - The effect of reboxetine on sympathetic neuroeffector transmission in rabbit isolated carotid artery was examined. Reboxetine (10-8-3 x 10-6 M) and cocaine (10-6-3 x 10-5 M), but not desipramine (10-8-3 x 10-7 M), increased contractions evoked by electrical field stimulation. At higher concentrations, reboxetine (10 4 M), cocaine (3 x 10-4 M), and desipramine (3 x 10-7-10-5 M) inhibited the neurogenic contractions. The enhancement seen with reboxetine and cocaine was partially reversible, whereas the inhibition was readily reversible. Reboxetine (10-7 M) and cocaine (10-5 M) prevented the inhibitory action of bretylium (10-6 M). Reboxetine (10-8-10-5 M), desipramine (10-7-10-4 M), and cocaine (10-6-10-5 M) increased the stimulation-evoked [3H]norepinephrine release. Pargyline (5 x 10 4 M) augmented the facilitatory effect of reboxetine (3 x 10-9-10-6 M) and cocaine (10-7-3 x 10-5 M). Reboxetine (10-8-10-6 M), desipramine (10-8-10-6 M), and cocaine (3 x 10-8-10-5 M) reduced the [3H]norepinephrine (10-8 M) uptake. Reboxetine (10-7 M) and cocaine (10-5-2 x 10-4 M) enhanced the contractions evoked by phenylephrine and norepinephrine. Higher concentrations of reboxetine antagonized the contractions. Reboxetine (10-5-6 x 10-5 M) antagonized the contractions evoked by potassium. The contractions evoked by tyramine (3 x 10-6 10-3 M) was reduced by reboxetine (3 x 10-8-10-6 M) and by cocaine (10-7-10-5 M). We conclude that reboxetine inhibits the membrane amine pump (uptake-1) in the terminals of postganglionic adrenergic neurons in a cocaine-like manner. PMID- 12754323 TI - Management of inflammatory neuropathies. PMID- 12754325 TI - Focal peripheral neuropathies. PMID- 12754324 TI - The diabetic neuropathies: types, diagnosis and management. PMID- 12754326 TI - Diagnosis and treatment of inflammatory muscle diseases. PMID- 12754327 TI - The neuromuscular junction disorders. PMID- 12754328 TI - Evaluating muscle symptoms. PMID- 12754329 TI - Systematic reviews to help guide clinical practice in neuromuscular disease. PMID- 12754330 TI - Axonal injury in multiple sclerosis. PMID- 12754331 TI - How common is dementia with Lewy bodies? PMID- 12754332 TI - Felix Guyon 1831-1920. PMID- 12754333 TI - Internet resources for neurologists. AB - This fourth and final review in the JNNP internet series summarises the essential internet resources for adult and paediatric clinical neurology, neuroradiology, and neurophysiology. This article is freely available on the JNNP website (www.jnnp.com), where-within seconds-the complete list of recommended websites can be easily downloaded and incorporated into your web browser as a Bookmark/Favorite file. The further progress of clinical neurology on the world wide web will be monitored in JNNP Neuronline fillers and JNNP Neurology in Practice supplements. PMID- 12754335 TI - Event based and time based prospective memory in Parkinson's disease. AB - BACKGROUND: Patients with Parkinson's disease have been reported to have retrospective memory impairment, while prospective memory, which is memory for actions to be performed in the future, has not yet been investigated. OBJECTIVE: To investigate the prospective memory of patients with Parkinson's disease. METHODS: Twenty Parkinson's disease patients and 20 age matched normal controls were given event based and time based prospective memory tasks. In the event based prospective memory task, the subject was asked to perform an action whenever particular words were presented. In the time based prospective memory task, the subject was asked to perform an action at certain times. RESULTS: The Parkinson's disease patients were impaired on the event based prospective memory task but not on the time based prospective memory task. The impairment of the Parkinson's disease patients on the event based prospective memory task was not the result of their forgetting the content of the prospective memory instructions, but the result of their failure to retrieve it spontaneously when the target words appeared. CONCLUSIONS: These results suggest that event based prospective memory is impaired in patients with Parkinson's disease, presumably relating to frontal lobe dysfunction. PMID- 12754336 TI - Using the SF-36 measure to compare the health impact of multiple sclerosis and Parkinson's disease with normal population health profiles. AB - OBJECTIVE: To examine the relative impact of two chronic neurological disorders, multiple sclerosis and Parkinson's disease, by comparing patients' scores on the medical outcomes study 36-item short form health survey (SF-36) with the health profile of the United Kingdom population norms. METHODS: 638 people representing the full spectrum of multiple sclerosis and 227 patients with Parkinson's disease were studied. Health status was measured by the SF-36. Scores for the eight health domains were compared after controlling for age, sex, disease duration, mobility, social class, ethnicity, education, marital status, and employment status. RESULTS: People with multiple sclerosis and those with Parkinson's disease had significantly worse health than the general population on all eight domains measured by the SF-36. The relative impact of multiple sclerosis and Parkinson's disease were similar, but multiple sclerosis resulted in poorer scores on physical functioning and better scores in mental health. People with mild multiple sclerosis who walked without an aid also had significantly worse scores in all dimensions than the general UK population. CONCLUSIONS: The results highlight the need for further research into aspects of health measured by the SF 36. Nevertheless, generic measures that are applicable across multiple diseases may fail to address clinically important aspects of the impact of specific disorders. PMID- 12754337 TI - Alzheimer's disease: differences in technetium-99m HMPAO SPECT scan findings between early onset and late onset dementia. AB - OBJECTIVE: To compare the HMPAO SPECT cerebral perfusion patterns in early and late onset Alzheimer's disease. METHODS: Twenty patients with early onset disease (<65 years) and 44 patients with late onset disease (>65 years) were studied. All patients fulfilled NINCDS-ADRDA clinical criteria and had details of disease severity and length of history at the time of imaging. Technetium-99m HMPAO SPECT brain scans were acquired on a multi-detector gammacamera and analysed visually and with statistical parametric mapping (SPM99). RESULTS: Patients with early onset disease had significantly greater posterior cortical association area involvement whereas those with late onset disease had significantly greater medial temporal hypoperfusion. These findings were unchanged after controlling for disease severity and length of illness. DISCUSSION: These functional imaging findings of the differences between early and late onset Alzheimer's disease are supported by published findings that include histopathological and clinical evidence; namely late onset patients tend to present with the characteristic involvement of the medial temporal lobes producing marked memory loss whereas early onset patients present with predominant posterior cortical association area involvement. These age related findings should be borne in mind when clinically diagnosing, and interpreting functional brain imaging studies in, patients with suspected Alzheimer's disease. PMID- 12754338 TI - Dementia with Lewy bodies according to the consensus criteria in a general population aged 75 years or older. AB - OBJECTIVE: To estimate the prevalence of dementia with Lewy bodies (DLB) according to the consensus criteria in a general population aged 75 years or older. METHODS: The "Kuopio 75+ study" is a population based health survey focused on the clinical epidemiology of dementia and functional capacity among elderly subjects aged 75 years or older. On 1 January 1998, a random sample of 700 subjects was drawn from a total population born before 1 January 1923, living in the city of Kuopio, northeast Finland (n = 4518). The study subjects underwent a structured interview and clinical examination. RESULTS: 601 elderly subjects (86% of the random sample) were examined. A dementia disorder was diagnosed in 137-a prevalence of 22.8% (95% confidence interval 19.4% to 26.2%). The prevalence of DLB was 5.0% (3.2% to 6.7%), comprising 22% of all demented subjects. Probable DLB was diagnosed in 20 subjects (3.3% (1.9% to 4.8%)), and possible DLB in 10 (1.7% (0.6% to 2.7%)). The prevalence of Alzheimer's disease was 10.6% (47% of all demented subjects), of vascular dementia, 5.3% (23%), and of other types of dementing disorders, 1.8% (8%). CONCLUSIONS: In a general population aged 75 years and older, the prevalence of a disorder fulfilling the diagnostic criteria of DLB is half that of Alzheimer's disease and the same as for vascular dementia. PMID- 12754339 TI - Neuroendocrine changes in patients with acute space occupying ischaemic stroke. AB - OBJECTIVE: To evaluate neuroendocrine changes in critical care patients with acute space occupying hemispheric stroke. METHODS: 22 patients with acute space occupying hemispheric stroke were studied (mean age 57.7 years; five women, 17 men). Plasma levels of prolactin, thyrotropin (TSH), total thyroxine (T4), free thyroxine (FT4), and total triiodothyronine (T3) were measured on admission and thereafter on days 3, 5, 7, and 9. Cortisol and ACTH levels were analysed at 8.00, 16.00, and 24.00 hours each day. A TRH stimulation test with measurements of TSH and prolactin was done on day 3. RESULTS: Nine patients underwent decompressive craniectomy and nine were treated with moderate hypothermia. All patients received vasopressor drugs because of arterial hypotension. Plasma ACTH and cortisol values were abnormally low despite systemic hypotension and acute systemic illness, and remained low throughout the observation period. The diurnal rhythm of cortisol was not preserved. Prolactin levels increased during the observation period, and were well above normal on day 9. Thyroid function was slightly suppressed until day 7. TRH stimulation of plasma TSH and prolactin was low. CONCLUSIONS: Patients with an acute space occupying cerebral infarct show profound neuroendocrine changes. The central regulation of adrenal and thyroid function and prolactin release is impaired, which may compromise the clinical course of affected patients and have implications for therapeutic management. PMID- 12754341 TI - Abnormalities on diffusion weighted magnetic resonance imaging performed several weeks after a minor stroke or transient ischaemic attack. AB - OBJECTIVES: Diffusion weighted brain imaging (DWI) is used in acute stroke, and also shows an acute ischaemic lesion in most transient ischamic attack (TIA) patients scanned acutely. However, it may also be useful in identifying subacute ischaemic lesions in patients with minor stroke or TIA who present several weeks after symptom onset. This study investigated the sensitivity and the observer reproducibility of DWI in cerebral TIA and minor ischaemic stroke patients scanned more than two weeks after the last symptomatic event. METHODS: Consecutive patients underwent magnetic resonance imaging (T2, DWI, ADC). The presence of clinically appropriate lesions was assessed by two independent observers, and related to the type of presenting event, the NIH score, persistence of symptoms and signs, and the time since the presenting event. RESULTS: 101 patients (53 men) were scanned at a median time of 21 days (IQR=17 28) after symptom onset. Reproducibility of the assessment of DWI abnormalities was high: interobserver agreement =97% (kappa=0.94, p<0.0001); intraobserver agreement =94% (kappa=0.88, p<0.0001). DWI showed a clinically appropriate ischaemic lesion in 29 of 51 (57%) minor stroke patients, and in 7 of 50 (14%) TIA patients. The independent predictors of a positive DWI scan were presentation with minor stroke versus TIA (p=0.009) and increasing NIH score (p=0.009), but there was no difference between patients presenting 2-4 weeks compared with >4 weeks after symptom onset. In minor stroke patients, the presence of a clinically appropriate lesion was associated with persistent symptoms (63% versus 36%; p=0.12) and signs (64% versus 33%, p=0.06) at the time of scanning. CONCLUSIONS: DWI shows a clinically appropriate ischaemic lesion in more than half of minor stroke patients presenting more than two weeks after the symptomatic event, but only in a small proportion of patients with TIA. The persistence of lesions on DWI is closely related to markers of severity of the ischaemic event. These results justify larger studies of the clinical usefulness of DWI in subacute minor stroke. PMID- 12754340 TI - Aetiological diagnosis of brain abscesses and spinal infections: application of broad range bacterial polymerase chain reaction analysis. AB - OBJECTIVE: To evaluate the usefulness of the broad range bacterial rDNA polymerase chain reaction (PCR) method combined with DNA sequencing in the aetiological diagnosis of intracranial or spinal infections in neurosurgical patients. METHODS: In addition to conventional methods, the broad range bacterial PCR approach was applied to examine pus or tissue specimens from cerebral or spinal lesions in patients treated in a neurosurgical unit for a clinical or neuroradiological suspicion of bacterial brain abscess or spondylitis. RESULTS: Among the 44 patients with intracranial or spinal lesions, the final diagnosis suggested bacterial disease in 25 patients, among whom the aetiological agent was identified in 17. A causative bacterial species was identified only by the rDNA PCR method in six cases, by both the PCR methodology and bacterial culture in six cases, and by bacterial culture alone in five. All samples in which a bacterial aetiology was identified only by the PCR approach were taken during antimicrobial treatment, and in three patients the method yielded the diagnosis even after >/= 12 days of parenteral treatment. One case also identified by the PCR approach alone involved a brain abscess caused by Mycoplasma hominis, which is not readily cultured by routine methods. CONCLUSIONS: In patients with brain abscesses and spinal infections, the broad range bacterial rDNA PCR approach may be the only method to provide an aetiological diagnosis when the patient is receiving antimicrobial treatment, or when the causative agent is fastidious. PMID- 12754342 TI - Intra-arterial thrombolysis in 24 consecutive patients with internal carotid artery T occlusions. AB - OBJECTIVES: To determine the safety, efficacy, and predictors of favourable outcome of intra-arterial thrombolysis in acute stroke attributable to internal carotid "T"occlusion METHODS: The authors analysed 24 consecutive patients with T occlusions of the internal carotid artery treated by local intra-arterial thrombolysis using urokinase. RESULTS: The median baseline National Institutes of Health Stroke Scale was 19. The average time from symptom onset to treatment was 237 minutes. Four patients (16.6%) had a favourable (modified Rankin Scale score (mRS 70 mm Hg). Fifty three patients admitted from 1997 to 2000 were treated using ICP/CPP management, but in this second group CPP was also increased as individually required to raise the P(ti)O(2) above 1.33 kPa (10 mm Hg) (P(ti)O(2) guided group). RESULTS: Cerebral hypoxic phases with P(ti)O(2) values below 1.33 kPa occurred significantly less often in the P(ti)O(2) guided group. P(ti)O(2) values were higher over the whole monitoring period. No statistical differences could be observed in outcome at six months, despite a positive trend in the P(ti)O(2) guided group. CONCLUSIONS: Cerebral hypoxic events can be reduced significantly by increasing cerebral perfusion pressure as required. To show a clear beneficial effect of P(ti)O(2) guided cerebral perfusion pressure management on outcome, a multicentre randomised trial needs to be undertaken. PMID- 12754348 TI - Cerebrovascular pressure reactivity is related to global cerebral oxygen metabolism after head injury. AB - BACKGROUND: After head injury, impaired cerebrovascular autoregulation has been associated with abnormally high or low cerebral blood flow. The physiological relevance of cerebral blood flow levels is difficult to assess in these patients, whose cerebral metabolic rate for oxygen (CMRO(2)) is known to be abnormal. Investigation of these relations requires quantitative measures of cerebral blood flow and CMRO(2), to allow assessment of oxygen supply and demand relations. OBJECTIVES: To investigate the relation between dysautoregulation and global cerebral oxygen metabolism following head injury. METHODS: Using positron emission tomography, global cerebral blood flow, CMRO(2), and oxygen extraction fraction were determined in 22 patients who were investigated in 26 examinations on days 1 to 11 (mean (SD), 3.5 (2.3)) after head injury. Cerebrovascular pressure reactivity was assessed using a pressure reactivity index, calculated as the moving linear correlation coefficient between mean arterial blood pressure and intracranial pressure. Outcome was assessed six months after injury using the Glasgow outcome scale. RESULTS: Low CMRO(2) was associated with disturbed pressure reactivity (inverse function, R(2) = 0.21, p = 0.018) and there was a correlation between disturbed pressure reactivity and oxygen extraction fraction (quadratic function, R(2) = 0.55, p = 0.0001). There was no significant relation between pressure reactivity and cerebral blood flow. An unfavourable outcome was associated with disturbed pressure reactivity. There was no significant relation between outcome and CMRO(2) or oxygen extraction fraction. CONCLUSIONS: There is a close relation between dysautoregulation and abnormal cerebral metabolism but not blood flow. Further studies are needed to determine whether metabolic dysfunction is a result of or a cause of disturbed pressure reactivity, and to establish if there is a relation between cerebral oxygen metabolism and outcome. PMID- 12754349 TI - Vestibular activation by bone conducted sound. AB - OBJECTIVE: To examine the properties and potential clinical uses of myogenic potentials to bone conducted sound. METHODS: Myogenic potentials were recorded from normal volunteers, using bone conducted tone bursts of 7 ms duration and 250 2000 Hz frequencies delivered over the mastoid processes by a B 71 clinical bone vibrator. Biphasic positive-negative (p1n1) responses were recorded from both sternocleidomastoid (SCM) muscles using averaged unrectified EMG. The best location for stimulus delivery, optimum stimulus frequency, stimulus thresholds, and the effect of aging on evoked response amplitudes and thresholds were systematically examined. Subjects with specific lesions were studied. Vestibular evoked myogenic potentials (VEMP) to air conducted 0.1 ms clicks, 7 ms/250-2000 Hz tones, and forehead taps were measured for comparison. RESULTS: Bone conducted sound evoked short latency p1n1 responses in both SCM muscles. Ipsilateral responses occurred earlier and were usually larger. Mean (SD) p1 and n1 latencies were 13.6 (1.8) and 22.3 (1.2) ms ipsilaterally and 14.9 (2.1) and 23.7 (2.7) ms contralaterally. Stimuli of 250 Hz delivered over the mastoid process, posterosuperior to the external acoustic meatus, yielded the largest amplitude responses. Like VEMP in response to air conducted clicks and tones, p1n1 responses were absent ipsilaterally in subjects with selective vestibular neurectomy and preserved in those with severe sensorineural hearing loss. However, p1n1 responses were preserved in conductive hearing loss, whereas VEMP to air conducted sound were abolished or attenuated. Bone conducted response thresholds were 97.5 (3.9) dB SPL/30.5 dB HL, significantly lower than thresholds to air conducted clicks (131.7 (4.9) dB SPL/86.7 dB HL) and tones (114.0 (5.3) dB SPL/106 dB HL). CONCLUSIONS: Bone conducted sound evokes p1n1 responses (bone conducted VEMP) which are a useful measure of vestibular function, especially in the presence of conductive hearing loss. For a given perceptual intensity, bone conducted sound activates the vestibular apparatus more effectively than air conducted sound. PMID- 12754350 TI - p53, mdm2, EGFR, and msh2 expression in paired initial and recurrent glioblastoma multiforme. AB - BACKGROUND: The clinical course of glioblastoma multiforme is characterised by invasive growth and regular recurrence. Many genetic alteration have been identified in the genesis of the disease. However, information about immunohistochemical expression in recurrent lesions is sparse. OBJECTIVES: To determine (1) whether the p53/mdm2/EGFR/msh2 expression pattern differs in initial v recurrent glioblastoma multiforme; (2) whether a possible change in expression correlates with prognostic variables (progression-free survival time, total survival time); and (3) whether chemotherapy in addition to surgery and radiotherapy influences the p53/mdm2/EGFR/msh2 expression profile. METHODS: 27 patients were studied. They met the following criteria: histologically confirmed diagnosis of glioblastoma multiforme (WHO IV); total tumour resection at initial craniotomy; at least one re-craniotomy for glioblastoma multiforme recurrence; age 21 years or older. All underwent radiotherapy of at least 54 Gy, and 17 received additional chemotherapy. Immunohistochemical staining of initial tumours and recurrences was done with the following monoclonal antibodies: anti-p53 (DO 1), anti-mdm2 (IF-2), anti-EGFR (H11), and anti-msh2 (AB-1). RESULTS: In comparison with the initial tumour, recurrent lesions were characterised by reduced expression of p53 (p < 0.0001) and msh2 (p = 0.0012), while the numbers of mdm2 (p = 0.02), EGFR (p < 0.0001), and msh2 positive specimens (p < 0.0001) were reduced. Chemotherapy was associated with reduced msh2 expression (p < 0.0001). Immunohistochemical variables were not associated with patient survival. CONCLUSIONS: There are significant differences in the p53/mdm2/EGFR/msh2 expression patterns in initial v recurrent glioblastoma multiforme. There may be interactions between chemotherapy and changes in the msh2 expression. PMID- 12754351 TI - Normal jugular bulb oxygen saturation. AB - BACKGROUND: Normal values of the jugular bulb oxygen saturation were obtained in 1942 and in 1963. Correct catheter positioning was not confirmed radiologically. OBJECTIVES: To replicate the measurements during angiographic catheterisation of the jugular bulb. METHODS: Oxygen saturation in the jugular bulb (SjO(2)), inferior petrosal sinus (SipsO(2)), and internal jugular vein was bilaterally measured in 12 patients with Cushing's syndrome undergoing selective bilateral catheterisation of the inferior petrosal sinus. In addition, data from the two old series were reanalysed for comparison. RESULTS: SjO(2) values (44.7%) were significantly lower than in the two old series, particularly concerning the normal lower limit (54.6% and 55.0% respectively). Comparative analysis suggests that contamination with the extracerebral blood of the facial veins and inferior petrosal sinuses was responsible for falsely high SjO(2) values in the two old series. CONCLUSIONS: The normal lower SjO(2) limit is lower than previously recognised. This may have practical implications for treating severe head trauma patients. PMID- 12754352 TI - Sir David Ferrier MD, FRS. PMID- 12754354 TI - Coexistence of CADASIL and Alzheimer's disease. AB - Cerebral autosomal dominant arteriopathy with subcortical infarcts and leucoencephalopathy (CADASIL) is caused by point mutations in the Notch3 gene. Presenilins are proteins involved in the cleaving of both Notch and the amyloid precursor protein (APP). In cases of early onset Alzheimer's disease mutations of the presenilin genes (PSEN 1 and PSEN 2) and APP can be found. A 64 year old patient with CADASIL (R169C-mutation) is reported, who, in addition to subcortical infarcts and granular osmiophilic deposits, had numerous senile plaques and neurofibrillary tangles on pathological examination. Mutations in the APP, PSEN1, and PSEN2 genes were not identified. Neuropathological findings of Alzheimer's disease may be found in CADASIL patients. PMID- 12754353 TI - Systemic infection, interleukin 1beta, and cognitive decline in Alzheimer's disease. AB - Activated microglia, the resident macrophages of the brain, are a feature of Alzheimer's disease. Animal models suggest that when activated microglia are further activated by a subsequent systemic infection this results in significantly raised levels of interleukin 1beta within the CNS, which may in turn potentiate neurodegeneration. This prospective pilot study in Alzheimer's disease subjects showed that cognitive function can be impaired for at least two months after the resolution of a systemic infection and that cognitive impairment is preceded by raised serum levels of interleukin 1beta. These relations were not confounded by the presence of any subsequent systemic infection or by baseline cognitive scores. Further research is needed to determine whether recurrent systemic infections drive cognitive decline in Alzheimer's disease subjects through a cytokine mediated pathway. PMID- 12754355 TI - Predictors of cognitive impairment in advanced Parkinson's disease. AB - OBJECTIVE: To investigate the cognitive profile of patients with idiopathic Parkinson's disease and to determine the demographic and medical variables that contribute to the cognitive outcome. DESIGN: Retrospective cohort analysis. METHODS: 100 patients with idiopathic Parkinson's disease were given a neuropsychological test battery investigating attention, memory, and visuospatial and executive functions. Test performance was compared against normative data, and linear regression determined significant predictors of cognitive impairment from a set of demographic and disease course variables. RESULTS: Frontal-type cognitive dysfunction was widespread in patients with advanced Parkinson's disease. Attention and memory were mildly to moderately impaired, whereas visuospatial function showed only subtle impairment. Older age and tremor at onset were significant predictors of poor cognitive performance. CONCLUSIONS: The observed cognitive impairment in patients with advanced Parkinson's disease is more than expected for normal aging. Although in apparent contrast with most previous research, reporting a greater risk of cognitive dysfunction in Parkinson's disease patients with predominant akinesia/rigidity, tremor at onset may be a marker for more widespread brain pathology that contributes to an increased risk of cognitive impairment. PMID- 12754356 TI - Hypertrophic olivary degeneration following pontine haemorrhage: hypertensive crisis or cavernous haemangioma bleeding? AB - The clinical and magnetic resonance (MR) features of hypertrophic olivary degeneration are described, along with a rare but treatable cause of this entity pontine cavernous haemangioma. Hypertrophic olivary degeneration occurs after focal lesions to the dentato-rubro-olivary pathway, typically following a pontine haemorrhage involving the ipsilateral central tegmental tract, the contralateral superior cerebellar peduncle, or the dentate nucleus. Clinically, there is palatal myoclonus and an uncontrollable tremor, presumably caused by loss of inhibitory control. On MR imaging, hypertrophic olivary degeneration is characterised by a non-enhancing T1 isointense, T2 hyperintense enlargement confined to the olivary nucleus. Typically, haemorrhages following a hypertensive crisis are responsible for hypertrophic olivary degeneration. However, in the three reported cases, imaging findings within the former bleeding cavity suggested a cavernous haemangioma as the source of the haemorrhage. PMID- 12754357 TI - Hemicraniectomy for large middle cerebral artery territory infarction: outcome in 19 patients. AB - BACKGROUND: Large space-occupying middle cerebral artery infarction accounts for 10-15% of all supratentorial infarctions and carries a mortality of 50% to 80%. Hemicraniectomy may be useful when optimal medical management has failed. METHODS: Between June 1997 and June 2000, 19 patients who fulfilled the clinical and imaging criteria for large middle cerebral artery infarction underwent hemicraniectomy because of impending herniation despite best medical therapy. The National Institute of Health Stroke Scale (NIHSS) assessed neurological status on admission and at one week after surgery. At 3 month follow up, The Barthel Index (BI) and Rankin Scale (RS) were used to assess the functional outcome among survivors. RESULTS: There were 15 males and 4 females with a mean age of 46.5 years (range 27-76 years). Ten patients (53%) had dominant hemisphere stroke. The mean interval between stroke onset and surgery was 60.3 hours (range 20-103 hours). The mean NIHSS score before surgery was 20.5 (range 17-26) and 10.5 (range 6-22) after surgery. One patient (5.2%) died due to post-operative meningitis. At follow up, mean BI was 56.4 (range 25-90) and RS revealed severe handicap in 4 patients (21%). Patients under 50 years of age had a significantly better outcome with mean BI of 60.7 as compared to only 41.3 (p=<0.048) in older patients. Speech function, especially comprehension improved in all patients with dominant hemisphere infarction. CONCLUSION: These findings add to previous studies suggesting hemicraniectomy may be a useful procedure in patients with large middle cerebral artery territory infarction. The functional outcome is good in younger patients. A randomised controlled trial is required to substantiate these findings. PMID- 12754358 TI - Mononeuritis multiplex in diabetes mellitus: evidence for underlying immune pathogenesis. AB - Four patients with type 2 diabetes mellitus developed mononeuritis multiplex subacutely. Sural nerve biopsies showed multifocal axonal loss in all patients, with epineurial perivascular inflammation affecting small calibre vessels in three. Three patients improved with immunotherapy. These observations suggest that mononeuritis multiplex in diabetes may be caused by an immune mediated vasculopathy and that it is pathogenetically akin to the more common and better recognised diabetic amyotrophy. PMID- 12754359 TI - Coccidioidomycosis of the brain, mimicking en plaque meningioma. PMID- 12754361 TI - BOLD fMRI activation induced by vagus nerve stimulation in seizure patients. AB - OBJECTIVE: To identify the cerebral activated regions associated with the vagus nerve stimulation in epilepsy patients. DESIGN: Blood oxygenation level dependent functional magnetic resonance imaging (BOLD fMRI) was employed to detect areas of the brain activated by vagus nerve stimulation in five patients with documented complex partial seizures. METHODS: Functional MRI was done on a GE 1.5T Echospeed horizon scanner. Before each patient entered the scanner, the vagal nerve stimulator was set to a specific ON-OFF paradigm so that the data could be analysed using a box-car type of design. The brains were scanned both anatomically and functionally. The functional images were corrected for head motion and co-registered to the anatomical images. Maps of the activated areas were generated and analysed using the brain mapping software, SPM99. The threshold for activation was chosen as p < 0.001. RESULTS: All patients showed activation in the frontal and occipital lobes. However, activation in the thalamus was seen only in the two patients with improved seizure control. CONCLUSIONS: BOLD fMRI can detect activation associated with vagus nerve stimulation. There may be a relation between thalamic activation and a favourable clinical outcome. PMID- 12754360 TI - The clinicopathological spectrum of Rosenthal fibre encephalopathy and Alexander's disease: a case report and review of the literature. AB - Alexander's disease is a leucodystrophy that usually presents in early childhood, but can infrequently arise in adults. It is characterised pathologically by megalencephaly, demyelination, and the presence of numerous Rosenthal fibres. Most cases have been shown to be due to mutations in the gene encoding glial fibrillary acidic protein. In rare instances, numerous Rosenthal fibres have been found at autopsy in patients who have suffered protracted debilitating systemic illnesses, some with associated brain stem signs, and in very rare instances in patients with no apparent neurological abnormality. The term "Rosenthal fibre encephalopathy" is used to distinguish these cases from those of Alexander's disease. We report the first case of Rosenthal fibre encephalopathy in a young man with AIDS, and review the literature. PMID- 12754362 TI - Risk of recurrence of cerebral venous and sinus thrombosis during subsequent pregnancy and puerperium. AB - OBJECTIVE: To assess the risk of recurrence of cerebral venous and sinus thrombosis (CVST) during subsequent pregnancy and puerperium in women with previous cerebral venous occlusive disease. METHODS: The authors retrospectively studied the relapse rate of CVST and the incidence of extracerebral venous thrombosis during subsequent pregnancies in 39 women (from 125 patients with CVST) who suffered a CVST at childbearing age. RESULTS: Mean follow up was 10.25 years (range 1 to 20). Twenty two pregnancies and 19 births were observed in 14 women without evidence of either recurrence of CVST or extracerebral venous thrombosis. One pregnancy occurred during oral anticoagulation and was interrupted and two pregnancies ended with spontaneous abortions. Low dose heparin had been given during five pregnancies. CONCLUSIONS: The risk of recurrence for CVST during pregnancy seems to be low and these data do not justify a negative advice on pregnancy in women with previous CVST. Further studies are needed to evaluate the need for a prophylactic anticoagulation during pregnancy and puerperium. PMID- 12754364 TI - Spastic movement disorder: what is the impact of research on clinical practice? PMID- 12754363 TI - Mental illness in new neurological patients. AB - OBJECTIVE: To determine the prevalence of psychiatric disorders in new neurological inpatients and outpatients, and examine whether they are recognised, treated, or referred to psychiatric consultation. METHODS: 198 consecutive patients referred for the first time to a neurologist were studied using a two phase design. ICD-10 psychiatric diagnoses were established by means of the SCAN (Schedules for Clinical Assessment in Neuropsychiatry). RESULTS: The overall prevalence of current mental disorders was 55.1% (95% CI: 46.2 to 63.8), and 65.0% (95% CI: 56.1 to 73.0) had at least once in their life had a psychiatric disorder. The most frequent current diagnoses were somatoform disorders (33.8%, (95% CI: 25.9 to 42.7%)), followed by phobias (21.8%; 95% CI: 15.3 to 30.0), substance use disorders (13.3%; 95% CI: 8.3 to 20.6) and depression/dysthymia (14.4; 95% CI: 9.1 to 21.8). The psychiatric morbidity markedly declined with increasing age. Compared with 63.5% of the women, 46.4% of the men had a psychiatric disorder. Substance use disorders were more frequent in men than women (p=0.002). Patients with a psychiatric disorder were more frequently seen in the outpatients' clinic than those without. The neurologists detected 14%-40% of the cases, 16.9% were in treatment, and only 4.6% were referred to mental health care. CONCLUSION: Psychiatric disorders, in particular somatoform disorders, are extremely common in neurological patients, especially in young and middle aged patients, outpatients, and women. The results call for more research on mental illness' impact on care and outcome in neurological patients. PMID- 12754365 TI - Central pontine myelinolysis temporally related to hypophosphataemia. PMID- 12754366 TI - Intracranial hypotension after chiropractic manipulation of the cervical spine. PMID- 12754367 TI - Clinical and electrophysiological improvement of adrenomyeloneuropathy with steroid treatment. PMID- 12754368 TI - Acute anterior radiculitis associated with West Nile virus infection. PMID- 12754369 TI - An Italian family affected by Nasu-Hakola disease with a novel genetic mutation in the TREM2 gene. PMID- 12754370 TI - A case of possible autoimmune bilateral vestibulopathy treated with steroids. PMID- 12754371 TI - Head injury outcome prediction in the emergency department: a role for protein S 100B? PMID- 12754372 TI - Subthalamic deep brain stimulation for advanced Parkinson's disease: all that glitters is not gold. PMID- 12754373 TI - Screening for variant Creutzfeldt-Jakob disease. PMID- 12754374 TI - Adrenergic regulation of clock gene expression in mouse liver. AB - A main oscillator in the suprachiasmatic nucleus (SCN) conveys circadian information to the peripheral clock systems for the regulation of fundamental physiological functions. Although polysynaptic autonomic neural pathways between the SCN and the liver were observed in rats, whether activation of the sympathetic nervous system entrains clock gene expression in the liver has yet to be understood. To assess sympathetic innervation from the SCN to liver tissue, we investigated whether injection of adrenaline/noradrenaline (epinephrine/norepinephrine) or sympathetic nerve stimulation could induce mPer gene expression in mouse liver. Acute administration of adrenaline or noradrenaline increased mPer1 but not mPer2 expression in the liver of mice in vivo and in hepatic slices in vitro. Electrical stimulation of the sympathetic nerves or adrenaline injection caused an elevation of bioluminescence in the liver area of transgenic mice carrying mPer1 promoter-luciferase. Under a light dark cycle, destruction of the SCN flattened the daily rhythms of not only mPer1, mPer2, and mBmal1 genes but also noradrenaline content in the liver. Daily injection of adrenaline, administered at a fixed time for 6 days, recovered oscillations of mPer2 and mBmal1 gene expression in the liver of mice with SCN lesion on day 7. Sympathetic nerve denervation by 6-hydroxydopamine flattened the daily rhythm of mPer1 and mPer2 gene expression. Thus, on the basis of the present results, activation of the sympathetic nerves through noradrenaline and/or adrenaline release was a factor controlling the peripheral clock. PMID- 12754375 TI - Gastrointestinal stromal tumors in a mouse model by targeted mutation of the Kit receptor tyrosine kinase. AB - Oncogenic Kit mutations are found in somatic gastrointestinal (GI) stromal tumors (GISTs) and mastocytosis. A mouse model for the study of constitutive activation of Kit in oncogenesis has been produced by a knock-in strategy introducing a Kit exon 11-activating mutation into the mouse genome based on a mutation found in a case of human familial GIST syndrome. Heterozygous mutant KitV558Delta/+ mice develop symptoms of disease and eventually die from pathology in the GI tract. Patchy hyperplasia of Kit-positive cells is evident within the myenteric plexus of the entire GI tract. Neoplastic lesions indistinguishable from human GISTs were observed in the cecum of the mutant mice with high penetrance. In addition, mast cell numbers in the dorsal skin were increased. Therefore KitV558Delta/+ mice reproduce human familial GISTs, and they may be used as a model for the study of the role and mechanisms of Kit in neoplasia. Importantly, these results demonstrate that constitutive Kit signaling is critical and sufficient for induction of GIST and hyperplasia of interstitial cells of Cajal. PMID- 12754376 TI - Programming peptidomimetic syntheses by translating genetic codes designed de novo. AB - Although the universal genetic code exhibits only minor variations in nature, Francis Crick proposed in 1955 that "the adaptor hypothesis allows one to construct, in theory, codes of bewildering variety." The existing code has been expanded to enable incorporation of a variety of unnatural amino acids at one or two nonadjacent sites within a protein by using nonsense or frameshift suppressor aminoacyl-tRNAs (aa-tRNAs) as adaptors. However, the suppressor strategy is inherently limited by compatibility with only a small subset of codons, by the ways such codons can be combined, and by variation in the efficiency of incorporation. Here, by preventing competing reactions with aa-tRNA synthetases, aa-tRNAs, and release factors during translation and by using nonsuppressor aa tRNA substrates, we realize a potentially generalizable approach for template encoded polymer synthesis that unmasks the substantially broader versatility of the core translation apparatus as a catalyst. We show that several adjacent, arbitrarily chosen sense codons can be completely reassigned to various unnatural amino acids according to de novo genetic codes by translating mRNAs into specific peptide analog polymers (peptidomimetics). Unnatural aa-tRNA substrates do not uniformly function as well as natural substrates, revealing important recognition elements for the translation apparatus. Genetic programming of peptidomimetic synthesis should facilitate mechanistic studies of translation and may ultimately enable the directed evolution of small molecules with desirable catalytic or pharmacological properties. PMID- 12754377 TI - The basic helix-loop-helix-PAS protein ARNT functions as a potent coactivator of estrogen receptor-dependent transcription. AB - The biological effects of estrogens are mediated by the estrogen receptors ERalpha and ERbeta. These receptors regulate gene expression through binding to DNA enhancer elements and subsequently recruiting factors such as coactivators that modulate their transcriptional activity. Here we show that ARNT (aryl hydrocarbon receptor nuclear translocator), the obligatory heterodimerization partner for the aryl hydrocarbon receptor and hypoxia inducible factor 1alpha, functions as a potent coactivator of ERalpha- and ERbeta- dependent transcription. The coactivating effect of ARNT depends on physical interaction with the ERs and involves the C-terminal domain of ARNT and not the structurally conserved basic helix-loop-helix and PAS (Per-ARNT-Sim) motifs. Moreover, we show that ARNT/ER interaction requires the E2-activated ligand binding domain of ERalpha or ERbeta. These observations, together with the previous role of ARNT as an obligatory partner protein for conditionally regulated basic helix-loop-helix PAS proteins like the aryl hydrocarbon receptor or hypoxia inducible factor 1alpha, expand the cellular functions of ARNT to include regulation of ERalpha and ERbeta transcriptional activity. ARNT was furthermore recruited to a natural ER target gene promoter in a estrogen-dependent manner, supporting a physiological role for ARNT as an ER coactivator. PMID- 12754378 TI - Copper chelation represses the vascular response to injury. AB - The induction of an acute inflammatory response followed by the release of polypeptide cytokines and growth factors from peripheral blood monocytes has been implicated in mediating the response to vascular injury. Because the Cu2+-binding proteins IL-1alpha and fibroblast growth factor 1 are exported into the extracellular compartment in a stress-dependent manner by using intracellular Cu2+ to facilitate the formation of S100A13 heterotetrameric complexes and these signal peptideless polypeptides have been implicated as regulators of vascular injury in vivo, we examined the ability of Cu2+ chelation to repress neointimal thickening in response to injury. We observed that the oral administration of the Cu2+ chelator tetrathiomolybdate was able to reduce neointimal thickening after balloon injury in the rat. Interestingly, although immunohistochemical analysis of control neointimal sections exhibited prominent staining for MAC1, IL-1alpha, S100A13, and the acidic phospholipid phosphatidylserine, similar sections obtained from tetrathiomolybdate-treated animals did not. Further, adenoviral gene transfer of the IL-1 receptor antagonist during vascular injury also significantly reduced the area of neointimal thickening. Our data suggest that intracellular copper may be involved in mediating the response to injury in vivo by its ability to regulate the stress-induced release of IL-1alpha by using the nonclassical export mechanism employed by human peripheral blood mononuclear cells in vitro. PMID- 12754379 TI - The specifics of small interfering RNA specificity. PMID- 12754380 TI - A single mutation in poliovirus RNA-dependent RNA polymerase confers resistance to mutagenic nucleotide analogs via increased fidelity. AB - Ribavirin is a nucleotide analog that can be incorporated by viral polymerases, causing mutations by allowing base mismatches. It is currently used therapeutically as an antiviral drug during hepatitis C virus infections. During the amplification of poliovirus genomic RNA or hepatitis C replicons, error frequency is known to increase upon ribavirin treatment. This observation has led to the hypothesis that ribavirin's antiviral activity results from error catastrophe caused by increased mutagenesis of viral genomes. Here, we describe the generation of ribavirin-resistant poliovirus by serial viral passage in the presence of increasing concentrations of the drug. Ribavirin resistance can be caused by a single amino acid change, G64S, in the viral polymerase in an unresolved portion of the fingers domain. Compared with wild-type virus, ribavirin-resistant poliovirus displays increased fidelity of RNA synthesis in the absence of ribavirin and increased survival both in the presence of ribavirin and another mutagen, 5-azacytidine. Ribavirin-resistant poliovirus represents an unusual class of viral drug resistance: resistance to a mutagen through increased fidelity. PMID- 12754381 TI - A fratricidal fungal prion. PMID- 12754382 TI - Biliary Complications of Pancreatic Necrosis. AB - Pancreatic necrosis has the potential to cause avariety of locoregional complications (1). This isbecause of the propensity of the necrotic tissue tospread far beyond the confines of the pancreas. Theproximity of the biliary tract to the pancreas makesit particularly vulnerable to damage by the inflammatory process, and though likely, there are only isolated case reports of involvement of the biliarytract through the necrotic process (2-5). This papershares our experience in managing six patients withbiliary complications of pancreatic necrosis. PMID- 12754383 TI - Inducible Nitric Oxide Synthase (iNOS) Immunoreactivity and Its Relationship to Cell Proliferation, Apoptosis, Angiogenesis, Clinicopathologic Characteristics, and Patient Survival in Pancreatic Cancer. AB - Human pancreatic adenocarcinoma remains oneof the most difficult cancers to treat, as evidencedby the minimal improvement in survival, even withthe addition of chemotherapy or radiation therapy. Epidemiological and experimental studies indicatethat the risk of developing pancreatic cancer may bethe result of the combined actions of environmentalfactors and endogenous promoting agents (1,2).Recently, much attention has been given to endogenousfactors, which appear to be directly responsiblefor the growth, spread, and invasion of tumorcells. The identification of such endogenous factorsshould lead to an understanding of the processes oftumor cell progression and metastasis, and also providenew strategies for developing agents that specificallysuppress these processes. PMID- 12754384 TI - Prognostic Significance of the Labeling of Adnab-9 in Pancreatic Intraductal Papillary Mucinous Neoplasms. AB - First described as a distinct entity in the early1980s (1), pancreatic ductal neoplasms with mucinhypersecretion have been increasingly recognized.This motivated the World Health Organization(WHO) reclassification proposal in 1996 (2), which separated them from mucinous cystic neoplasms.These tumors are now (3) termed intraductal papillarymucinous neoplasms (IPMN). Despite a burgeoningvolume of recent literature devoted to thiscondition, little is known of the pathogenesis ofIPMN,which is believed to constitute 10% of mucinproducingpancreatic tumors and 1% of pancreaticcancers (4). IPMN presents diagnostic and therapeuticchallenges to the clinician because it representsa histologic spectrum of morphology, from thebenign adenoma to invasive carcinoma. The initialhistological and morphological features of IPMN oftenunderestimate its invasive potential (5), and may notaccurately predict survival. Conversely, it may oftenbe difficult to differentiate benign from malignantlesions (5-7). In the most advanced stages, when aninvasive carcinoma is present, IPMN can be indistinguishablefrom common pancreatic ductal cancer(PC), yet with aggressive surgical management, theprognosis for patients with IPMN is far better (6,7).The availability of a prognostic indicator, independentfrom the pathological stage, may help to directtherapy. PMID- 12754385 TI - Primary Tuberculosis of the Pancreas Mimicking a Pancreatic Tumor. AB - Tuberculosis of the pancreas is very rare, and isgenerally associated with miliary tuberculosis (1).The correct diagnosis is often missed if there is noevidence of tuberculosis in other organs (2). Therefore,it can present to the clinician as a difficult diagnosticdilemma. We present an extremely rare case of primary pancreatictuberculosis that was initially diagnosed aspancreatic tumor, in a non-immunocompromisedpatient who did not have miliary tuberculosis. PMID- 12754386 TI - Is Glucagonoma of the Pancreas a Curable Disease? AB - Glucagonomas are rare tumors that arise from thepancreatic islet cell of Langerhans. Because of theuncommon occurrence of these tumors, the naturalhistory is not well defined and has largely been basedon case reports or small series. Less than 250 casesof glucagonomas have been reported in the Englishliterature. To our knowledge, the largest single institutionalexperience is from the Mayo Clinic where 21 cases were reported (1). We report one of thelargest series of 12 patients with a diagnosis ofglucagonomas treated at our institution. PMID- 12754387 TI - Effects of Mast-Cell Stabilization in Cerulein-Induced Acute Pancreatitis in Rats. AB - Acute pancreatitis is characterized by inflammationof the pancreas that ranges from mild inflammationto severe necrotizing pancreatitis (1-3).Several factors can initiate a cascade of pathologic events that results in AP (4). A key feature in thispathogenesis is inflammatory reaction. However,causes of exaggerated inflammatory reaction are notwell understood (4,5). Since the pathogenesis of pancreatitisis still unclear, most of the treatment modalitiesare largely supportive and independent of thepathogenesis (1,2). PMID- 12754388 TI - Pancreatic Polypeptide Secretion in Patients with Chronic Pancreatitis and After Pancreatic Surgery. AB - Pancreatic polypeptide (PP) is a 36-amino acidpolypeptide with a molecular weight of 4200 (1). PP is mainly produced in the head of the pancreas in adistinct cell type both within the islets and scatteredamong the exocrine parenchyma (2,3). PP release isunder neural and hormonal control. Ingestion of nutrients,cholinergic neural activation and infusion ofgastro-intestinal peptides (especially CCK) stimulatePP release (1,4,5). It has been clearly shown that PPsecretion is under vagal cholinergic control since thePP response to a meal or CCK infusion is bluntedduring vagal cholinergic blockade with atropine orfollowing truncal vagotomy (1,6). PMID- 12754389 TI - In Vivo Effect of Pancreatic Phospholipase A2 on the Arachidonic Acid Cascade. AB - Acute pancreatitis causes various systemic complicationssuch as shock, renal failure, respiratory failure, disseminated intravascular coagulation, and eventually,multiple organ dysfunction syndrome. These systemic complications are responsible for the highmortality rates of acute pancreatitis. Thus, an understandingof their pathogenesis is imperative to reducethe mortality of acute pancreatitis. PMID- 12754390 TI - Distribution of Nitric Oxide Synthase and Secretory Role of Exogenous Nitric Oxide in the Isolated Rat Pancreas. AB - Exocrine pancreatic secretion is controlled for themost part by the autonomic neurons and by I and Stype enteroendocrine cells (1). In addition to the classic neurotransmitters and various neuropeptides that are apparently produced and stored in the pancreaticnerve cells (2), there is morphological evidencethat nitric oxide (NO) containing (nitrergic) nervesare also present in the pancreas (3). Nitric oxide synthase(NOS) containing perikarya and fibers havebeen shown in the pancreas of several species usingenzymehistochemical and immunohistochemicalmethods (4). In the present study the nitrergic nervesof the rat pancreas were demonstrated by NOSimmunohistochemistry. PMID- 12754391 TI - Expression of the IIIc Variant of FGF Receptor-1 Confers Mitogenic Responsiveness to Heparin and FGF-5 in TAKA-1 Pancreatic Ductal Cells. AB - Human pancreatic cancer is a devastating diseasewith a poor prognosis (1). Although the exact reasonsfor the aggressive nature of this disorder are unknown, certain observations have pointed to theimportant role of growth factors in its pathobiology(2). Many of these cancers frequently overexpressfibroblast growth factors (FGFs) (3). FGF signalingis mediated through four high-affinity tyrosine kinasereceptors, termed fibroblast growth-factor receptors(FGFRs) (3,4). The extracellular domain of FGFRsis usually composed of three immunoglobulin (Ig) like domains (I-III), a transmembrane region followedby a juxtamembrane domain, and a splittyrosine kinase catalytic domain. Several isoformsof FGFR-1, -2, and 3 have been identified, some of which exhibit different ligand-binding properties(3,5). Alternative splicing of the second half of Igdomain III of FGFR 1 results in three receptor variants,termed IIIa, IIIb, and IIIc. The IIIa splice variantyields a secreted receptor that is devoid of anysignaling capacity (5). The expression of the IIIbvariant is generally restricted to epithelial cell types,whereas the expression of the IIIc variant is restrictedto mesenchymal cell types (6-11). PMID- 12754392 TI - Neurofibromatosis-1 and Ampullary Gangliocytic Paraganglioma Causing Biliary and Pancreatic Obstruction. AB - Gangliocytic paragangliomas (GPs) are exceedingly rare tumors that arise in close proximity of the papillaof Vater. Nevertheless, jaundice is an uncommon presenting symptom, reported in only 3 of 125 casesdescribed in the literature to date, with gastrointestinal bleeding being more common.Association between GPs and neurofibromatosis 1 (NF-1), described in two patients, may be more thancasual, being the frequency in normal population 1:3000.We report an additional case of this association, presenting for the first time with simultaneous obstructionof the common biliary duct, and of the main pancreatic duct. Previous cases of GPs with jaundice and/orNF-1 are reviewed, and updating of the histogenesis presented. PMID- 12754393 TI - Cavernous Pancreatic Ductal Ectasia with Smooth Muscle Proliferation Causing Recurrent Acute Pancreatitis. AB - Pancreatic cystic lesions have various etiologies,including pseudocyst (inflammatory cyst), retentioncyst, congenital cyst, and neoplastic cyst (1).Recently, we experienced a case with a uniquepancreatic cyst-like lesion causing recurrent acutepancreatitis. This patient had multiple cystic dilatations of branch pancreatic ducts surrounded by proliferationof smooth muscle tissue, with neither neoplastic nor inflammatory changes. To our knowledge, there are no previous reports of such a case. PMID- 12754394 TI - Portal Vein Thrombosis and Pancreatic Failure. AB - We report a case of a child with portal vein thrombosis presenting with protein losing enteropathy. Helater developed exocrine and endocrine pancreatic failure. This association has not been reported before. PMID- 12754395 TI - Pancreatic Arteriovenous Malformation Associated with Duodenal Ulcer and H.Pylori Infection. AB - Pancreatic arteriovenous malformation (PAVM) is a rare condition that may cause duodenal ulcer. A36-yr-old man with PAVM associated with duodenal ulcer and H.Pylori infection is described. The patienthad recurrent episode of upper abdominal pain despite healed ulcer and H.Pylori eradication.The preoperative diagnosis was confirmed by computed tomography and the patient was treated with apancreatoduodenectomy. Histological examination of the resected pancreas revealed a pancreatic arteriovenousmalformation involving the adjacent duodenal wall. PMID- 12754396 TI - Small-Bowel Obstruction Secondary to Subcutaneous Small-Bowel Entrapment: A Late Complication of Laparostomy for Necrotizing Pancreatitis. AB - Laparostomy is a well recognized strategy for the management of patients who have necrotizing pancreatitisand may require multiple re-intervention. The open wound can be left to heal through a process ofgranulation and contraction. This article describes intestinal obstruction secondary to entrapment of a loopof small bowel within the cicatrix of the contracting cutaneous scar. An awareness of the potential for entrapmentof the small bowel in the healing scar is critical for clinicians using laparostomy in the managementof acute necrotizing pancreatitis. PMID- 12754397 TI - The Lustgarten Foundation for Pancreatic Cancer Research 3rd Annual Scientific Conference: "Pancreatic Cancer: From Genes to Treatment" AB - MEETING NOTES: The Lustgarten Foundation for Pancreatic Cancer Research 3rd Annual Scientific Conference: "Pancreatic Cancer: From Genes to Treatment" PMID- 12754398 TI - The Latest in Pancreatic Cancer Research: Lustgarten Foundation Awardees Present Their Findings. AB - On June 13 and 14, 2001, the Lustgarten Foundationheld its third annual Pancreatic Cancer ScientificConference in Baltimore, Maryland. Thisconference was entitled "Pancreatic Cancer: FromGenes to Treatment" and was held in conjunctionwith The Johns Hopkins University School of Medicine.As it had for the previous two years, the conferenceenabled pancreatic cancer scientists fromaround the world to meet and discuss new conceptsin pancreatic cancer research. The conference alsoincluded a lecture series and poster presentations oncurrent topics in pancreatic cancer research and treatment.These presentations gave researchers andhealth care professionals the opportunity to sharetheir ideas, with the ultimate goal of finding a curefor pancreatic cancer. PMID- 12754399 TI - Adjuvant and Neoadjuvant Therapies of Pancreatic Cancer: A Review. AB - The survival of patients diagnosed with pancreatic cancer is dismal. Few patients on initial presentation aresuitable for surgical resection. This has prompted clinical studies with chemotherapy and/or radiotherapydesigned either to increase the number of patients eligible for surgery (neoadjuvant therapy) or to prolong thesurvival of patients who had undergone surgery (adjuvant therapy). None of these studies may at this time beconsidered definitive. Wherever possible, patients felt eligible for neoadjuvant or adjuvant therapy should beentered on clinical trials. Where this is not possible, clinicians should exercise their best judgment in offeringthis type of treatment to pancreatic cancer patients under their care. PMID- 12754400 TI - Toxicity and Efficacy of Concurrent Gemcitabine and Radiotherapy for Locally Advanced Pancreatic Cancer. AB - Gemcitabine has been demonstrated to be a potentradiosensitizer in the laboratory and in the clinic (1-7)and has proven clinical systemic activity to pancreaticcancer. Responses to systemic gemcitabine inpatients with metastatic pancreatic adenocarcinomahave been documented in phase I, phase II, and phaseIII clinical settings (8,9). Moreover, a recent randomizedtrial of gemcitabine vs 5 FU as first-linetherapy in patients with advanced pancreatic adenocarcinomademonstrated a modest median survivalbenefit (4.41 vs 5.65 mo,p= 0.0025) for those patientswho received gemcitabine compared to those whoreceived 5-FU (10). In addition, gemcitabine wasshown to improve cancer-related symptoms and performancestatus as assessed by a quantitative clinicalbenefit scale in both untreated and previouslytreated patients with metastatic adenocarcinoma ofthe pancreas (10,11). Based on these data, the FDAapproved gemcitabine as a first line agent for patientswith advanced adenocarcinoma of the pancreas. PMID- 12754401 TI - WR-2721 Reduces Intestinal Toxicity from Concurrent Gemcitabine and Radiation Treatment. AB - The success of radiotherapy as a single treatmentmodality is limited by several factors, including tumor radioresistance due to hypoxia within a tumormass, efficient DNA repair mechanisms, cellularrecovery during the time between radiation fractions,and distant failure from occult tumor cells outsidethe radiation field. For these reasons, radiotherapyis being increasingly combined with other treatmentmodalities, especially chemotherapy. The resultsfrom this type of combined modality treatment haveshown increased local tumor control rates anddecreased distant metastasis (1-4). PMID- 12754402 TI - Mouse Models of Metastatic Pancreatic Adenocarcinoma. AB - Pancreatic adenocarcinoma is a deadly disease. Its etiology is unknown, and metastatic disease kills themajority of patients who have it. Effective prevention is clearly the ultimate goal for eradicating this diseaseprovided that the effects of environmental and genetic elements on pancreatic cancer development arefully understood. Currently, it appears that the control of pancreatic cancer metastasis is of immediateurgency. Fulfillment of this difficult task relies on knowledge of the cellular and molecular biology of metastasis.The use of relevant animal models will help define each aspect of this complicated process. PMID- 12754403 TI - A Novel, Clinically Relevant Animal Model of Metastatic Pancreatic Adenocarcinoma Biology and Therapy. AB - In this study, we report a metastatic model of Panc02 murine pancreatic adenocarcinoma. Parental Panc02cells were orthotopically implanted into the pancreas of syngeneic C57BL/6 mice. Tumor cells were isolatedfrom liver micrometastases 90 d after tumor implantation and established as a culture (Panc02-H1).The Panc02-H1 cells were then implanted into the pancreas of mice. Liver metastases were then collectedand established as Panc02-H2 cells. This process was repeated until the Panc02-H7 cell line was established.These cells were extremely aggressive after implantation as manifested by progressive growth in the pancreas,peritoneal dissemination, and distant metastasis to multiple organs, including the liver and lungs.Moreover, Panc02-H7 cells expressed the inducible nitric oxide synthase gene at a very low level in cultureand produced highly vascularized tumors having a large number of infiltrating macrophages. Collectively,this model system should be a valuable tool for investigating the molecular mechanisms governing pancreaticcancer growth and metastasis and exploring potential treatment modalities for this disease. PMID- 12754404 TI - Expression of Heparin-Binding Epidermal Growth Factor-like Growth Factor in Pancreatic Adenocarcinoma. AB - Heparin-binding epidermal growth factor-likegrowth factor (HB-EGF) belongs to the EGF familyand was originally identified in the conditionedmedium of a human histiotic lymphoma cell line, U937 (1). This protein is synthesized as a preproformof 208 amino acids (2), is expressed as amembrane-anchored HB-EGF, and is released as asoluble form of 87 amino acids after being processedby protease (1,2). HB-EGF binds to the EGF receptor(EGF-R) in competition with EGF (3), and exogenoussoluble HB-EGF is known to stimulateDNA synthesis in primary rat hepatocytes in a dosedependentmanner (4). PMID- 12754405 TI - Adenosquamous Carcinoma of the Pancreas: Successful Treatment with Extended Radical Surgery, Intraoperative Radiation Therapy, and Locoregional Chemotherapy. AB - Adenosquamous carcinoma of the pancreas is arare tumor and accounts for only 2-4% of tumorsof the exocrine pancreas (1,2). These tumors are histologicallycharacterized by adenomatous cell populationsmixed with varying amounts of keratinizedsquamous cell elements (1,3). PMID- 12754406 TI - Emergency Pancreaticoduodenectomy with Delayed Reconstruction for Bleeding: A Life Saving Procedure. AB - Emergency pancreaticoduodenectomy with delayed reconstruction can be performed as a life saving procedurein case of massive bleeding uncontrolled with conventional hemostatic techniques. The authors reportherein the case of a 39-yr-old patient with an acute episode of chronic pancreatitis-induced massive bleedingsuccessfully treated by this unorthodox technique.The concept of damage control surgery with abbreviated laparotomy and planned reconstruction couldbe useful in selected cases outside the trauma setting. PMID- 12754407 TI - Resectable Pleomorphic Giant Cell Carcinoma of the Pancreas. AB - We present a 72-yr-old woman who underwent surgical resection of a large pancreatic tumor. On preoperativeimaging, an intratumoral cavity containing necrotic tissue was noted, mimicking the appearance of acystic tumor. There was no invasion of adjacent organs nor distant metastasis, and histopathologic examinationrevealed the tumor to be a giant cell carcinoma. Postoperatively, the patient has been followed for 1.5 yrwithout evidence of recurrence. Giant cell carcinoma generally is associated with a poor prognosis, and patientsusually die within months despite intensive multimodality therapy. Some patients with giant cell carcinomas,however, achieve long-term survival when invasion of adjacent organs and distant metastases are absent. Surgicalresection is the appropriate treatment for tumors with these favorable characteristics. PMID- 12754408 TI - Gold compound auranofin inhibits IkappaB kinase (IKK) by modifying Cys-179 of IKKbeta subunit. AB - Antirheumatic gold compounds have been shown to inhibit NF-kappaB activation by blocking IkappaB kinase (IKK) activity. To examine the possible inhibitory mechanism of gold compounds, we expressed wild type and mutant forms of IKKalpha and beta subunits in COS-7 cells and determined the effect of gold on the activity of these enzymes both in vivo and in vitro. Substitution of Cys-179 of IKKbeta with alanine (C179A) rendered the enzyme to become resistant to inhibition by a gold compound auranofin, however, similar protective effect was not observed with an equivalent level of IKKalpha (C178A) mutant expressed in the cells. Auranofin inhibited constitutively active IKKalpha and beta and variants; IKKalpha (S176E, S180E) or IKKbeta (S177E, S181E), suggesting that gold directly cause inhibition of activated enzyme. The different inhibitory effect of auranofin on IKKalpha (C178A) and IKKbeta (C179A) mutants indicates that gold could inhibit the two subunits of IKK in a different mode, and the inhibition of NF-kappaB and IKK activation induced by inflammatory signals in gold-treated cells appears through its interaction with Cys-179 of IKKbeta. PMID- 12754409 TI - Rapid uptake of oxidized ascorbate induces loss of cellular glutathione and oxidative stress in liver slices. AB - The observation that ascorbate known to retain pro-oxidant properties induces cell death in a number of immortal cell lines, led us to examine its mechanism and whether it is involved in oxidative stress injury in such asocorbate-enriched tissue cells as hepatocytes. In rat liver homogenates, higher concentrations (1 and 3 mM) of ascorbate suppressed lipid peroxide productions but lower concentrations (0.1 and 0.3 mM) did not. In contrast to the homogenate, ascorbate increased lipid peroxide production in liver slices in a concentration dependant manner. Iso-ascorbate, the epimer of ascorbate did not cause an increase the oxidative stress in liver slices. This differential effect between homogenates and liver slices implies that cellular integrity is required for ascorbate to induce oxidative stress. Wortmannin, an inhibitor of the GLUT (glucose transporter) thought to transport dehydroascorbate into cells, inhibited [(14)C] ascorbate uptake and suppressed oxidative stress in liver slices. Wortmannin suppressed that [(14)C]-ascorbate uptake by GLUT following oxidation to [(14)C]dehydroascorbate. Taken together, these observations support our hypothesis that ascorbate is oxidized to dehydroascorbate by molecular oxygen in solution (i.e., plasma and culture medium) which is then carried into hepatocytes (via a GLUT) where it is reduced back to ascorbate causing oxidative stress. PMID- 12754410 TI - Association between interleukin 6 promoter variants and chronic hepatitis B progression. AB - Interleukin 6 (IL6) plays an essential role in the regulation of immune response to chronic disease. In this study, the three known single nucleotide polymorphisms (SNPs) in the IL6 promoter region were genotyped in a large chronic hepatitis B cohort to evaluate the effects of IL6 promoter variants. The single base extension method was used for this genotyping. Haplotypes were constructed by the three SNPs in IL6. Allele frequencies were compared for; i) patients with chronic hepatitis (CH) and chronic carriers vs. chronic hepatis patients with clinical evidence of liver cirrhosis (LC) (i.e., portal hypertension), ii) cirrhotic patients with hepatocellular carcinoma (HCC) vs. without HCC by logistic regression, and iii) with respect to the time intervals from the onset of infection to HCC. Results were analyzed by Cox relative hazard analysis on the assumption that all the patients were infected during early infancy. The frequencies of each SNP were 0.002 (IL6-597 G>A), 0.25 (IL6-572 C>G) and 0.002 (IL6-174 G>C), respectively, in the Korean population (n = 1,046). No significant associations were detected between IL6-572 C>G and chronic hepatitis B outcome in this study; i.e., LC occurrence on CH (OR = 0.16-1.27, P = 0.13- 0.71) and HCC occurrence on LC (OR = 1.04-1.23, P = 0.89-0.60) of heterozygotes and homozygotes for G allele in referent comparison to homozygotes for common allele (C/C genotype), and time interval to HCC (RH = 0.67-1.00; P = 0.14-0.99). In conclusion, there appeared to be no significant associations between IL6 promoter variants and disease outcome in chronic hepatitis B. PMID- 12754412 TI - Herba houttuyniae extract induces apoptotic death of human promyelocytic leukemia cells via caspase activation accompanied by dissipation of mitochondrial membrane potential and cytochrome c release. AB - Herba houttuyniae has been used as a constituent of herval medicine prescriptions for the treatment of inflammation, cancer, and other diseases. In the present study, we investigated the cellular effects of herba houttuyniae extract (HHE) and the signal pathways of HHE-induced apoptosis in HL-60 human promyelocytic leukemia cell line. HHE treatment caused apoptosis of cells as evidenced by discontinuous fragmentation of DNA, the loss of mitochondrial membrane potential, release of mitochondrial cytochrome c into the cytosol, activation of procaspase 9 and caspase-3, and proteolytic cleavage of poly(ADP-ribose) polymerase. Pretreatment of Ac-DEVD-CHO, caspase-3 specific inhibitor, or cyclosporin A, a mitochondrial permeability transition inhibitor, completely abolished HHE-induced DNA fragmentation. Together, these results suggest that HHE possibly causes mitochondrial damage leading to cytochrome c release into cytosol and activation of caspases resulting in PARP cleavage and execution of apoptotic cell death in HL-60 cells. PMID- 12754411 TI - Arsenic trioxide-induced apoptosis is independent of stress-responsive signaling pathways but sensitive to inhibition of inducible nitric oxide synthase in HepG2 cells. AB - Arsenic trioxide (As(2)O(3)) has been found to be remarkably effective in the treatment of patients with acute promyelocytic leukemia (APL). Although evidences for the proapoptotic activity of As(2)O(3) have been suggested in leukemic and other solid cancer cells, the nature of intracellular mechanisms is far from clear. In the present study, we investigated As(2)O(3) affect on the stress responsive signaling pathways and pretreatment with antioxidants using HepG2 cells. When treated with micromolar concentrations of As(2)O(3), HepG2 cells became highly apoptotic paralleled with activation of caspase-3 and members of mitogen-activated protein kinases (MAPKs) including extracellular signal regulated kinase (ERK) and c-jun NH(2)-terminal kinase (JNK) but not p38 MAP kinase. However, inhibition of each kinase activity failed to inhibit apoptosis by As(2)O(3). Addition of n-acetyl cysteine (NAC) or diphenyleneiodonium (DPI) effectively protected cells from apoptosis and significantly lowered As(2)O(3) induced activation of caspase-3. However, neither NAC nor DPI was able to effect ERK or JNK activation induced by As(2)O(3). Guanidinoethyldisulfide dihydrochloride (GED) and 2-ethyl-2-thiopseudourea (ETU), known inhibitors of the inducible nitric oxide synthase (iNOS), also suppressed the apoptotic activity of As(2)O(3). These results suggest that As2O3 induces caspase-mediated apoptosis involving a mechanism generating oxidative stress. However, activation of some stress-responsive signaling pathways by As(2)O(3) may not be the major determinant in the course of apoptotic processes. PMID- 12754414 TI - Base excision repair synthesis of DNA containing 8-oxoguanine in Escherichia coli. AB - 8-oxo-7,8-dihydroguanine (8-oxo-G) in DNA is a mutagenic adduct formed by reactive oxygen species. In Escherichia coli, 2,6-dihydroxy-5N formamidopyrimidine (Fapy)-DNA glycosylase (Fpg) removes this mutagenic adduct from DNA. In this report, we demonstrate base excision repair (BER) synthesis of DNA containing 8-oxo-G with Fpg in vitro. Fpg cut the oligonucleotide at the site of 8-oxo-G, producing one nucleotide gap with 3' and 5' phosphate termini. Next, 3' phosphatase(s) in the supernatant obtained by precipitating a crude extract of E. coli with 40% ammonium sulfate, removed the 3' phosphate group at the gap, thus exposing the 3' hydroxyl group to prime DNA synthesis. DNA polymerase and DNA ligase then completed the repair. These results indicate the biological significance of the glycosylase and apurinic/ apyrimidinic (AP) lyase activities of Fpg, in concert with 3' phosphatase(s) to create an appropriately gapped substrate for efficient BER synthesis of DNA containing 8-oxo-G. PMID- 12754413 TI - Expression of hOGG1 protein during differentiation of HL-60 cells. AB - Human 8-oxo-G-DNA glycosylase 1 (hOGG1) is a DNA glycosylase to cleave 8-oxo-7,8 dihydroguanine (8-oxo-G), a mutagenic DNA adduct formed by oxidant stresses. Here, we examined hOGG1 protein expression and repair activity to nick a DNA strand at the site of 8-oxo-G during differentiation of hematopoietic cells using HL-60 cells. Overall expression of hOGG1 protein was increased during granulocytic differentiation of HL-60 cells induced by DMSO and monocytic differentiation by vitamine D(3). Greater level of hOGG1 protein was expressed in DMSO-treated cells. However, change in the DNA nicking activity was not in parallel with the change in hOGG1 protein expression, especially in PMA-treated cells. In PMA- treated cells, the level of hOGG1 protein was lowered, even though the DNA nicking activity was elevated, in a manner similar to the changes in serum- deprived HL-60 cells. These results indicate that hOGG1 expression change during differentiation of hematopoietic stem cells for adaptation to new environments. And the DNA cleaving activity may require additional factor(s) other than expressed hOGG1 protein, especially in apoptotic cell death. PMID- 12754415 TI - De novo mutations in sporadic deletional Duchenne muscular dystrophy (DMD) cases. AB - Dinucleotide repeat polymorphism based genetic analysis is a powerful approach to gain insight into rare genetic events like germline mosaicism and de novo mutations. The loss of heterozygosity of polymorphic dinucleotide loci at "deletional hotspot" of dystrophin gene can provide direct evidence of carrier status in female relatives of affected DMD patients with overlapped exonic deletions. We have used 4 STR loci of the central deletional hotspot of the dystrophin gene for genetic analysis in sporadic unrelated DMD families. Twenty nine mothers of sporadic deletional cases were analysed and their carrier status was determined. Eighteen of them showed heterozygosity in the deleted loci suggesting the occurrence of de novo mutations. In 9 cases, the carrier status was indeterminate while 2 showed germline mosaicism. Our observations reiterated the importance of STR analysis in determining the status of mothers of sporadic deletional DMD cases in order to provide proper genetic counselling. PMID- 12754416 TI - Activation of epidermal growth factor receptor is responsible for pervanadate induced phospholipase D activation. AB - Pervanadate, a complex of vanadate and H(2)O(2), has an insulin mimetic effect, and acts as an inhibitor of protein tyrosine phosphatase. Pervanadate-induced phospholipase D (PLD) activation is known to be dependent on the tyrosine phosphorylation of cellular proteins and protein kinase C (PKC) activation, and yet underlying molecular mechanisms are not clearly understood. Here, we investigated the signaling pathway of pervanadate-induced PLD activation in Rat2 fibroblasts. Pervanadate increased PLD activity in dose- and time- dependent manner. Protein tyrosine kinase inhibitor, genistein, blocked PLD activation. Interestingly, AG-1478, a specific inhibitor of the tyrosine kinase activity of epidermal growth factor receptor (EGFR) blocked not only the PLD activation completely but also phosphorylation of p38 mitogen-activated protein kinase (MAPK). However, AG-1295, an inhibitor specific for the tyrosine kinase activity of pletlet drived growth factor receptor (PDGFR) did not show any effect on the PLD activation by pervanadate. We further found that pervanadate increased phosphorylation levels of p38, extracellular signal-regulated kinase (ERK) and c Jun NH(2)-terminal kinase (JNK). SB203580, a p38 MAPK inhibitor, blocked the PLD activation completely. However, the inhibitions of ERK by the treatment of PD98059 or of JNK by the overexpression of JNK interacting peptide JBD did not show any effect on pervanadate-induced PLD activation. Inhibition or down regulation of PKC did not alter the pervanadate-induced PLD activation in Rat2 cells. Thus, these results suggest that pervanadate-induced PLD activation is coupled to the transactivation of EGFR by pervanadate resulting in the activation of p38 MAP kinase. PMID- 12754417 TI - Partial rescue of the Na+-Ca2+ exchanger (NCX1) knock-out mouse by transgenic expression of NCX1. AB - The null mutation of cardiac Na(+)-Ca(2+) exchanger (NCX1) gene in mice caused death of embryo in utero at embryonic day (ED) 9.0-9.5 and this embryonic lethality appears resulted from abnormal heart development. In the present study, we investigated whether transgenic re-expression of NCX1 in mutant cardiac myocytes could rescue these lethal defects. Transgenic mice expressing the canine NCX1 in a cardiac specific manner were bred into the NCX1 knock-out background but did not prevent the fetal lethality associated with the NCX1 null allele. However, the NCX1 knock-out embryos with an NCX1 transgene survived with heart beatings until ED 10.5 which was one day longer than the survival of the NCX1 knock-out embryos (ED 9.5). At ED 10.5, however, the partially rescued NCX1 embryos might have succumbed to the lack of an organized vasculature in the yolk sacs. The placental labyrinth layer was reduced in size and largely avascular. The transgenic re-expression of NCX1 rescued heart beatings and survived longer, but was still insufficient for the mice to be completely rescued. Importantly, NCX1 was observed to express in the yolk sac and the placenta of wild type mice. The results suggest that defects in extra-embryonic compartments are causal to the lethality, and that NCX1 may play an important role in establishing vascularization in extra-embryonic tissues. PMID- 12754418 TI - Epigallocatechin gallate, a constituent of green tea, suppresses cytokine-induced pancreatic beta-cell damage. AB - Cytokines produced by immune cells infiltrating pancreatic islets have been implicated as one of the important mediators of beta-cell destruction in insulin dependent diabetes mellitus. In this study, the protective effects of epigallocatechin gallate (EGCG) on cytokine-induced beta-cell destruction were investigated. EGCG effectively protected IL-1beta and IFN-gamma-mediated cytotoxicity in insulinoma cell line (RINm5F). EGCG induced a significant reduction in IL-1beta and IFN-gamma-induced nitric oxide (NO) production and reduced levels of the inducible form of NO synthase (iNOS) mRNA and protein levels on RINm5F cells. The molecular mechanism by which EGCG inhibited iNOS gene expression appeared to involve the inhibition of NF-kappaB activation. These findings revealed EGCG as a possible therapeutic agent for the prevention of diabetes mellitus progression. PMID- 12754419 TI - [Is atopic dermatitis an allergic disease?]. PMID- 12754420 TI - [Oral therapy for atopic dermatitis in year 2000. Questionnaire Study to Boarding Members of Japanese Society for Allergology]. AB - Questionnaires were sent to boarding members of Japanese Society for Allergology (response rate 65.2%; 49 dermatologists [Derma] and 101 pediatricians [Pedia] ) to ask the order of choice in therapy attitudes, in the oral in comparison to the local therapy and in the oral therapy only. As for therapy attitudes, "suppress inflammation" (Infla), "reduce staphylococcal colonization" (Staph), "inhibit scratching" (Scrat), "guard skin from irritation" (Guard), "avoid allergen" (Avoid), and "support psychologically" (Psych) were presented. The order of choice was Guard, Infla, Scrat and Avoid for the first four in both Derma and Pedia. The last 2 were Psych and Staph in Derma and vice versa in Pedia. However, the lowest adoption rate was 76.2% in Derma and 85.8% in Pedia. This suggests that wide therapy attitudes were taken in mind in the therapy of atopic dermatitis in the both specialites. The adoption rate was significantly (P>0.05) higher in Pedia than Derma with Staph, Avoid, Scrat and Psych. As for the oral versus local therapy, "antihistamine" (A-hist), "antiallergy" (A-aller), "sedative" (Sedat), "hypnotic" (Hypno), "corticosteroid hormone" (O-steroid) and "antibacterial drug" (A-bact) were presented for the oral therapy, and "skin care medicine" (SkinC), "NSAID" (t-NSAID), "corticosteroid ointment" (S-oint) and "disinfectant" (Disinf) were for the topical therapy. The adoption rate was in the order of SkinC as well as S-oint followed by A-aller and A-hist both in Derma and Pediat. These choices were over 60% in Derma and over 75% in Pediat. Disinf and t-NSAID followed these with over 50% of adoption rate in Pediat, while all others were 30.6% or less in Derma. Thus, in Derma 4 therapies (2 local and 2 oral) were thought to be the first rank therapy but in Pediat additional 2 therapies (Disinf, t-NSAID) were comparable to the first rank therapy in Derma. As for the oral therapy only, A-aller and A-hist were adopted at high rates both in Derma and Pediat, but all other therapies that included 5 additional therapies; "kampo drug", "antimycotic drug", "vitamine", "arachidonic acid cascade controlling drug" and "immunosuppressant" were adopted in only low rates. Experience of oral corticosteroid therapy was about three quarters in Derma but a quarter in Pediat. Conditions for prescribing oral corticosteroid were mostly "extreme severity" and "acute generalized aggravation" both in Derma and Pediat. About 40% of doctors both in Derma and Pediat who ever prescribed oral corticosteroid were afraid that corticosteroid might not be stopped when starting the therapy. However, only short-term oral corticosteroid therapy is described in the guidelines of this society for the treatment of atopic dermatitis (1995). It is hoped to standardized oral corticosteroid therapy for long-lasting extremely severe atopic dermatitis. PMID- 12754421 TI - [A case of anaphylaxis due to matsutake mushroom]. PMID- 12754422 TI - [A case of fixed eruption due to tonic water]. PMID- 12754423 TI - [A case of Sjogren syndrome accompanied by interstitial cystitis and interstitial pneumonia improved by suplatast tosilate and prednisolone]. PMID- 12754424 TI - [Scientific research and financing]. PMID- 12754425 TI - [Intestinal mucosa immunity and oral vaccines especially regarding E. coli vaccines: a review]. AB - The aim of this work was a review of literature with regard to the mucosal immunity, the oral vaccines and the bacterial lysates. The Gut Associated Lymphoid Tissue (GALT) include effector and inductive sites and is constituted by organized and diffuse tissues. GALT defends the integrity of the gut, inhibits the development of allergy and autoimmunity and induce a mucosal and systemic immune response against enteric antigens. Bacterial lysates are innocuous and can reduce the frequency and the seriousness of diarrhoea, mucosal infections and diverticulitis; they induce the production and the biologic activity of secretory IgA and cytokines. The DNA vaccines are able to induce a strong immune response; the oral vaccines formulated with bacterial adhesins can inhibit the entry of pathogens and oral antigens across the gut. The use of adjuvants can amplify the activity of the oral vaccines; an objective of the research is the discovery of potent adjuvants without remarkable toxicity. PMID- 12754426 TI - [Antibiotic therapy in the elderly: features and problems]. AB - Bacterial infections cause 30% of deaths in the elderly and are the most frequent cause of hospitalization in elderly patients. Diagnosis of infection can be difficult because aged patients may have neither fever nor leucocytosis; most patients present unusual symptoms such as changes in mental status. The clinician must be cognizant of the frequent noncompliance with drug regimens because lack of elderly adherence to a prescribed antibiotic therapy has the potential to result in treatment failure and to foster the emergence of drug-resistant bacteria. Elderly frequently are taking other drugs such as antiarrhythmics and antihypertensives; ignorance of potential antibiotic-drug interaction can result in ineffective treatment or enhanced toxicity. Aging is associated with changes in physiological processes; the age-related decline in renal function influences the excretion of some antibiotics (aminoglycosides, vancomycin, ofloxacin). The increased potential for toxicity of antimicrobial agents requires a careful drug selection as well as clinical and laboratory monitoring. The most frequent infections occurring in the elderly are pneumonia, urinary tract infection, intra abdominal infection and soft tissue infection; prevalence and incidence of bacterial meningitis, bacterial endocarditis and bacteraemia are increasing with a mortality rate of 20 to 40%. These bacterial infections have different microbial causes and require different therapeutic approaches according to sites involved, elderly's salient features and overall susceptibility of the bacteria in the ecosystem. Appropriate empirical antibiotic treatment reduces mortality also in bacteraemic old patients. PMID- 12754427 TI - [Pharyngotonsillitis caused by Streptococcus pyogenes: clinical and epidemiological aspects and resistance phenotypes towards macrolides]. AB - During 2001 we analyzed 1730 pharyngeal swabs for S. pyogenes (SGA): 1142 children (0-10 years old), 132 adolescent subjects (11-17 years old), and 456 adults (18 or more years old). 994 subjects (664 children, 85 adolescent ones, 245 adults) had acute pharyngotonsillitis. In this last group we observed 321 positivities (32.3 %) for SGA: 40.4 % among children, 24.7 % among adolescent people, 13.1 % among adults. The pharyngotonsillitis prevailed during winter and spring. The resistances (R) towards erythromycin were 27.7 % (89 cases), and among children 30.6 % (82 cases), towards clyndamicin 15.3 % (49 cases, and 17.2 %, 46 cases, among children), towards rokytamicin 11.8 % (38 cases, and 13.1 %, 35 cases, among children). These were the phenotypes of R to erythromycin: 25.8 % M-phenotype, 19.1 % inducible (iMLS), 55.1 % constitutive (cMLS); among children respectively 25.6 %, 18.3 % and 53.7 %. Increased resistances towards 16-C macrolides, increased resistances of cMLS to erythromycin, and the persistence of R to 14-C macrolides around 30 % are discussed. PMID- 12754428 TI - Have Ureaplasma urealyticum and Mycoplasma hominis infections any significant effect on women fertility? AB - Ureaplasma urealyticum and Mycoplasma hominis are known as sexually transmitted agents. U. urealyticum and M. hominis jeopardize male fertility. However, it is unclear whether these infections significantly contribute to female infertility. In this controlled-study we aimed to establish whether M. hominis and U. urealyticum are risk factors for female fertility and prevalence of infection from these agents in patients attending our infertility clinic. Total 96 married women enrolled in this prospective study; the infertile (study) group consisted of 50 women and fertile (control) group comprised 46 women. The patients were searched about the presence of U. urealyticum and M. hominis by a micro-liquid culture method. The samples were collected from endocervical area with a dacron swab. 28 of 50 (56%) and 18 of 46 (39%) women were evaluated as positive for U. urealyticum culture in the study and control groups respectively. M. hominis was cultured from 4 of 50 (8%) women in the study group as no positive result in controls. There were no statistically significant differences between the groups for both agents (p>0.05), but the higher prevalence of U. urealyticum in infertile women gives emphasis to evaluate these agents in patients that have no any other etiological factor for infertility. PMID- 12754429 TI - [Infective endocarditis in a series of 1527 autopsies: clinical-pathology correlations]. AB - The study of 1527 autopsies showed that in 2% of cases the cause of death was Infective Endocarditis (IE) (acute in the absolute majority), mostly not diagnosed due to fulminant disease or to the short hospital stay and in some cases not correctly treated. Sepsis was correctly diagnosed in 12 patients (9 of whom were i.v. drug addicts) and was not identified in 14 patients (12 of whom had acute disease) who were diagnosed as suffering from CNS disease or pneumonia. Systematic autoptic study of patients with or without IE appears to be a very useful method to determine correctly IE mortality. PMID- 12754430 TI - [Outpatient parenteral antibiotic therapy (OPAT) of diabetic foot infections with piperacillin/tazobactam]. AB - Treatment of diabetic foot infections (DFIs) represents an important challenge for surgeons, especially in light of the poor results achieved by traditional therapeutic approaches. In this study, the clinical and bacteriological efficacy of TZP for treatment of DFIs in 38 outpatients was evaluated. All patients (median age 63 yrs) were affected by DFIs to different degrees of severity according to Wagner's classification: degree 0, 7 pts; degree 1, 17 pts; degree 2, 10 pts; degree 3, 4 pts. Degree 0-1 infections underwent a 10-18 day course with TZP given i.m. (2.25 g bid); degree 2-3 infections were initially treated with TZP i.v. (4.5 g bid or tid). Some patients began treatment in hospital and after early discharge continued parenteral therapy at home; others were treated exclusively at home. Some pts, after a 5-7-day course of i.v. therapy switched to i.m. route. The average duration of antibiotic therapy was 28 days. At the end of treatment with TZP, some patients underwent a new treatment with oral coamoxi clav for 10-15 days. A bacteriological examination was done for all patients: ulcus (degree 1) and deep tissue (degree 2-3) swabs at the first surgical toilette. Clinical controls, medications, surgical toilettes and microbiological cultures were performed according to the degree of severity, extension of the lesion and response to treatment. All cultures were positive for polymicrobial infections (Staphylococcus spp, Enterococcus spp, Enterobacteriaceae, Pseudomonas spp). In 30/38 pts (79%) a complete resolution was observed; in 4 pts (10%) an improvement. DFIs require long term parenteral treatment, with wide spectrum antibiotics including Gram +, Gram - and anaerobes. OPAT represents a valid alternative to hospitalisation when the general conditions of the patient are stable, the infection is not too severe and complications are not present. TZP proved to be a good choice for treatment of diabetic foot infections that, due to its high safety, can be successfully utilized also in OPAT programmes PMID- 12754431 TI - [Acute hepatitis-associated pure red cell aplasia: a case report]. AB - After a holiday in Egypt, a 57-year-old caucasian woman developed acute hepatitis A. The illness seemed initially to have a benign course, with a decreasing trend of hepatic enzymes and an apparent recovery. Three weeks later a relapse occurred. Recurrence of symptoms and aminotransferase elevation were associated with severe anemia; a hyporegenerative anemia was diagnosed and all laboratory findings were consistent with pure red cell aplasia. The haematologic disorder was successfully treated with red cell transfusion and glucocorticoids. PMID- 12754432 TI - [World microbiological unification according to Woodrow Borah]. AB - Now that developments in communication media are turning the world into a "global village" and there are risks of global epidemics due to infectious agents spread by possible terrorist attacks, this article aims to remind us of what happened in the "New World", where in the space of a few decades the indigenous civilizations disappeared. When, following the geographical discoveries of the 15th, 16th and 17th centuries, millions of people met European sailors, soldiers and traders, the outbreak of pathocenosis was destructive--the intensity of this effect was directly related to the degree of isolation of the indigenous populations colonised by the Europeans. The author of this article recalls the Woodrow Borah thesis. Borah, a former History Professor at Berkeley University, elaborated a theory according to which the microbiological unification of the world is the prime cause of the genocide of the indigenous population in America and Oceania. The author emphasises that the events following the discovery of the American continent are very similar to the plague epidemic which started in 1348 in Europe. PMID- 12754443 TI - [Treatment of severe Gram-positive infections: current situation and new opportunities]. AB - METHICILLIN-RESISTANT STAPHYLOCOCCUS AUREUS (MRSA) INFECTIONS: A growing number of MRSA strains with an increased minimum inhibitory concentration (MIC) and intermediary susceptibility to glycopeptides (GISA) or vancomycin (VISA) are encountered in clinical practice. In patients on mechanical ventilation who develop acute pneumonia, it would appear appropriate to achieve a vancomycin concentration in serum and the lung 2 to 4 times above the MIC, while carefully monitoring the risk of toxicity. POSSIBLE SOLUTIONS: Better prevention of severe MRSA infections, a more rational use of glycopeptides, using a recycling scheme, i.e. altering prescriptions with antibiotics other than glycopeptides or using combinations. OTHER AGENTS CURRENTLY AVAILABLE: Several antibiotic classes can now be used to preserve the efficacy of glycopeptides: cotrimoxazole, quinupristine/dalfopristine and linezolide. Linezolide is the first compound of a new family of antibiotics called oxazolidinones which are active against aerobic and anaerobic Gram positive strains, particularly those exhibiting intermediary sensitivity or resistance to other antibiotics. Its pharmacokinetic properties are quite favorable. ANTIBIOTIC COMBINATIONS: In case of severe MRSA infection, antibiotics that can be combined with vancomycin include gentamycin, rifampicin, or fosfonycine. For GISA infections, vancomycin could be combined with a ss lactam, or quinupristine/dalfopristine. Combination with linezolide appears to be antagonistic. PMID- 12754442 TI - [Epidemiology of Gram-positive infections in France: changing resistance]. AB - CONTRIBUTION OF CLINICAL EPIDEMIOLOGY: Clinical epidemiology goes beyond simply counting cases of resistance. The goal is to collect clinical data contributing to an explication of epidemiological observations enabling a classification into 4 types of resistance. GRAM POSITIVE BACTERIA: Depending on the specialty, Gram positive bacteria are the cause of a variable percentage of clinical infections. In oncology and hematology, Gram-positive infections appear to be on the rise due to the increase in the number of coagulase-negative staphylococcal infections. The percentage appears to be unchanged in intensive care. Community-acquired streptococcal infections include a growing number caused by Streptococcus viridans, with the development of resistance and an aggravation of Streptococcus pneumoniae resistance. Nosocomial Gram-positive infections show that the increase in the number of meticillin-resistant Staphylococcus aureus infections is slowing down with the emergence of exceptional Staphylococcus aureus strains exhibiting intermediary resistance to glycopeptides. STREPTOCOCCUS VIRIDANS: Isolation of Streptococcus viridans is becoming increasingly frequent, particularly in immunodepressed patients or in patients with surgical conditions such as peritonitis. Several risk factors have been identified. In vitro glycopeptides and linezolide would have 100% activity against S. viridans. METICILLIN-RESISTANT STAPHYLOCOCCUS AUREUS (MRSA): Infections caused by MRSA strains are rather exceptional compared with simple colonization. All patients with suspected Staphylococcus aureus infections should not be treated as if they had a MRSA infection. STREPTOCOCCUS PNEUMONIA: Beta-lactame resistant pneumococci have shown some progression. PMID- 12754444 TI - [Contribution of linezolid to the treatment of nosocomial pneumonia]. AB - TWO PREREQUISITES: In order to avoid the emergence of bacterial resistance, the treatment of nosocomial infections must avoid unnecessary multiplication of antibiotics. Treatment must also be given early for these life-threatening infections. The presence of resistant and/or multiresistant strains is a common cause of antibiotic failure. EPIDEMIOLOGY: The frequency of methicillin-resistant Staphylococcus aureus (MRSA) strains involved in nosocomial pneumonia remains relatively moderate. It would appear logical to anticipate the presence of MRSA strains in patients given prior antibiotic therapy. EFFICACY OF LINEZOLIDE: Three clinical trials in patients with nosocomial pneumonia assessed the efficacy of linezolid. All three demonstrated an equivalent efficacy for linezolide and vancomycin. OTHER ADVANTAGES OF LINEZOLID: Besides its efficacy, linezolid has a good tolerance profile, particularly interesting for intensive care patients with multiple organ insufficiency. It can be given orally and can help reduce hospital stay, an important contribution to cost savings. PMID- 12754445 TI - [Contribution of linezolid to the treatment of complicated infections of the skin and soft tissues]. AB - THREE GROUPS OF INCREASING SEVERITY: According to the 2000 consensus conference, bacterial infections of the skin and soft tissues can be classified in three categories of increasing severity: bacterial dermo-hypodermal infections, bacterial dermo-hypodermal necrotizing infections, necrotizing fasciitis. The last two categories are medicosurgical emergency conditions requiring repeated surgical debridement and probabilistic antibiotic therapy. LINEZOLID: The clinical efficacy of linezolid in bacterial infections of the skin and soft tissues is comparable to that of oxacillin/dicloxacillin. Several arguments (frequent implication of Streptococcus pyogenes and Staphylococcus aureus, well adapted spectrum, changing resistance pattern of Gram-positive cocci, efficacy against associated bacteriemia, favorable kinetics and diffusion data in cutaneous tissues) argue in favor of the use of linezolid in infections of the skin and soft tissues PMID- 12754446 TI - [Is human cloning therapeutic?]. PMID- 12754447 TI - [What is expected of psychologists in palliative care mobile teams? Their role and missions]. AB - The present development of palliative care structures has led to the organisation, in institutes and also in the home, of multi-disciplinary teams destined that contribute to the care and accompanying of persons at the end of their life. The multiplication of these mobile teams invites us to gain further knowledge on the respective role of each professional and in particular that of the clinical psychologist in a function that is often ignored by the institutions. This study is aimed at providing a better definition of the role and missions of a psychologist working in a mobile team. The psychologists' clinical activity is specified with regard to interventions with the patients and their families and also their intervention among the medical and care professionals who look after these patients. The choice of working in a interdisciplinary group leads to a collaboration of quality and the dynamics of such collaboration guarantee the global approach of the patients in their physical, mental, social and spiritual suffering. PMID- 12754448 TI - [Medicinal iatrogenics in hospitals. A survey on a given day]. AB - OBJECTIVE: Medicinal iatrogenics are responsible for hospital admissions but also occur in hospitals. In view of the lack of knowledge, prevalence and nature of the adverse drug-related events (ADE) in the Bichat-Claude Bernard hospital group in Paris, and because of the potential severity of the latter, the Local drug committee has decided to develop a policy to manage these risks. METHOD: The first stage consisted in a transversal study on a given day in the departments in which patients are hospitalised for more than 24 hours, in order to assess the prevalence, severity and preventability of ADE and to search for factors of risk. RESULTS: 107 ADE were observed in 89 patients on the day of the survey (9.9% global prevalence of ADE [CI 95%: 8.8% - 11.0%]). Among the latter, 57 patients had exhibited at least one adverse event during their hospitalisation, i.e., a prevalence of 6.3% ([CI 95%: 4.7% - 7.9%] ). Two thirds of these patients were hospitalised in medical departments. These nosocomial ADE (nosocomial adverse drug events) were serious or severe in 73% of cases and 25% could have been avoided. The only clearly identified risk factor was the number of drugs prescribed. CONCLUSION: This review has drawn the attention of the medical and paramedical community to the need to define vigilance markers, and has provided some elements of response that should be further completed by a prospective cohort study. PMID- 12754449 TI - [Evaluation of discomfort and complications in a population of 18,102 patients overweight or obese patients]. AB - AIMS: The burden of disorders associated with overweight and obesity is a major public health problem. It is therefore important to better identify these concomitant disorders and how their frequencies vary with sex and age. METHODS: A survey was carried out during a 5 month-period from September 2001 to January 2002) among 4 727 general practitioners distributed throughout France in 18 102 patients with a body mass index (BMI)>25 kg/m2. The practitioners evaluated the presence of concomitant disorders using a closed questionnaire. The patients assessed global discomfort linked to overweight using an analog visual scale. Univariate and multivariate analyses of the concomitant disorders and self reported discomfort depending on age, gender and BMI were performed. RESULTS: The survey population comprised 66.8% of women (W) and 33.2% of men (M). Mean age was 48.0 +/- 13.2 years and mean BMI was 34.6 +/- 6.1, with no differences between the two sexes. The most frequent concomitant disorders were back pain (44.6%), hypertension (44.2%), dyslipidemia (39.9%), knee osteoarthritis (30.8%), lower limb edema (24.3%), hypersudation (23.8%), skin fold mycosis (22.8%) and type 2 diabetes (21.6%). In multivariate analyses, the distribution of these disorders varied with sex: hypertension, type 2 diabetes, dyslipidemia, and hypersudation were more frequent in men, whereas knee osteoarthritis, back pain, and skin fold mycosis were more frequent in women. The prevalence (odd ratio, OR) of back pain and dyslipidemia did not increase with higher BMI and the prevalence of back pain did not increase with age. Overall discomfort related to overweight was rated as 61.3 +/- 19.9 mm on a 0 to 100-mm scale. Discomfort was less marked in men, decreased with age and increased with BMI (and with the consultations in the Paris area). CONCLUSIONS: This study shows the complexity of relationships between concomitant diseases, overall discomfort, BMI, age and sex (in the population of overweight and obese patients) and should improve the management of such patients and their complications. PMID- 12754450 TI - [Arizonae subspecies Salmonella enterica urinary infection with confusional syndrome]. AB - INTRODUCTION: Infections due to subspecies arizonae Salmonella enterica are rare. These infections are transmitted by reptiles. We report the case of S. arizonae urinary tract infection that occurred in an immunocompetent woman. OBSERVATION: An 82 year-old woman was admitted for confusion. A urinary tract infection due to Salmonella arizonae was diagnosed. No neurological, iatrogenic or metabolic cause could explain the confusion. Treatment with ciprofloxacin was given. The confusion and the infectious syndrome disappeared. CONCLUSION: The prevalence of infections is probably underestimated because the digestive problems they generate are usually benign. PMID- 12754451 TI - [Abdominal actinomycosis. Three cases]. AB - BACKGROUND: Abdominal actinomycosis is a suppurating affection caused by a positive Gram germ, Actinomyces israelii. Manifestations of this rare disease are usually pseudotumoral syndromes leading to surgical exeresis. The diagnosis is obtained from the histologic report. CASES REPORT: We report three cases in all of which this disease presents with tumoral symptoms. The first affected the sigmoid, the second affected the mesentery and the last affected the greater omentum and abdominal wall. In no case, the diagnosis was done before surgery. In 2 of the cases reported here, the patients had an old, incontrolled intra-uterine device. DISCUSSION: From a review of the literature, we observed that the diagnosis of abdominal actinomycosis remains difficult. The radiologic findings are nonspecific. Actinomycosis is treated with prolonged antibiotics, and surgery is only needed in abdominal abscess and fistula. CONCLUSION: Theses observations illustrates the difficulties of the diagnosis of this rare unrecognized disease. PMID- 12754452 TI - [Regression of hepatic fibrosis physiopathological aspects and clinical reality]. AB - MANAGING THE RESPONSIBLE AGENT: Hepatic fibrosis with its end-point, cirrhosis, are the principle complications responsible for morbidity and mortality in chronic liver diseases. It is therefore important to address the question of whether these lesions can disappear, once installed in the liver. Regression can only occur when the agent responsible for the fibrosis (virus, alcohol, poison, iron, autoantibodies, etc) is eradicated or controlled. THE FORMS OF REGRESSION: Once the agent controlled, regression of fibrosis can either be spontaneous, a rare situation, although some bona fide cases of fibrosis or even cirrhosis reversion have been reported in the literature, or assisted by specific therapy. It is therefore necessary to take into consideration the development of new treatments based on enhanced knowledge of the mechanisms of fibrosis. THE ACTIVITY AND EFFICACY OF TREATMENTS: These treatments target one of the three following mechanisms: the blockade of hepatic stellate cell activation, enzymatic digestion of fibrous tissue and stimulation of liver cell regeneration. Although these treatments have shown efficacy on experimental models of fibrosis, to date, there are no published results formally confirming the efficacy and safety of these treatments in man. PMID- 12754453 TI - The genesis of cognitive and behavioral deficits in premature graduates of intensive care. AB - Increased survival of very low birth weight infants including those born at the cutting edge of viability is associated with substantial cognitive and behavioral deficits at follow-up that has extended into school age and adolescence. These problems have occurred as common in the presence or absence of neurosonographic abnormalities. Factors/events that may predispose to these problems include medical complications of prematurity i.e. chronic lung disease, recurrent episodes of apnea and bradycardia, transient hypothyroxinemia of prematurity, hyperbilirubinemia, nutritional deficiencies, medications used to treat such conditions i.e. glucorticoids, theophylline etc. and stress associated with prolonged hospitalization. With regard to the latter, attachment to multiple devices that limits infant provider interactions, high noise levels and constant light levels are considered to be of particular importance. Experimental evidence is presented that demonstrates the value of positive interactions between the subject and provider with regard to neurobehavioral outcome. Some suggested interventions include reducing noise levels and displacing it with music, modulating light exposure and enhancing infant parent interactions such as kangaroo care. Finally the important postnatal role of social influences on cognitive and behavioral outcomes is discussed. PMID- 12754454 TI - [Renal transplantation in the pediatric age]. AB - Renal transplantation is the optimal and preferred treatment for children with end-stage renal disease. Pediatric kidney transplantation results have improved significantly over the years and the actuarial survival of the children with renal transplantation has become excellent. These improvements are due to many factors, including better immunosuppressive regimens and therefore a decrease in acute rejection episodes and possible improvement of graft survival. The concentration of care in specialized pediatric transplantation centers allowed the improvement of kidney transplants also in children less than 6 years old. The same success is not always achieved in infants. The selection of the donor is another important factor. The survival rate of renal transplantation is better in case of living-related donors. Renal transplants performed from cadaveric donors <6 years of age have an actuarial survival lower than renal transplants from cadaver donors >6 years of age. Owing to the limited members of cadaveric kidneys available for transplants, also the donors <6 years old are sometimes a valuable resource. As far as HLA-matching and its relationship with renal transplant outcome is concerned, there are conflicting data, but important registers on adults and children show the positive relationship between histocompatibility matching and graft outcome. A major distinguishing feature of pediatric from adult renal recipients is the need for children to grow. It is well known that chronic renal insufficiency involves a growth failure. A functioning transplant may improve the growth, but a catch-up growth is rarely achieved. To overcome this problem many techniques, such as alternate-day steroid therapy, discontinuation of prednisone, the use of recombinant growth hormone, have been adopted. As to social rehabilitation, transplanted children attend the school and work more than dialyzed ones. PMID- 12754456 TI - [Guidelines: state of the art and peculiar aspects in pediatric emergency]. AB - In recent years pediatric guidelines have increasingly become part of clinical practice, in Italy too. Aim of the present work is, on the basis of a review of national and international literature, to focus on the Italian situation about the use of guidelines in pediatric emergency, with particular respect to methodology, correct use, and related risks and benefits. The developing of efficient guidelines is achieved in different steps: 1) identifying and refining the topic for guidelines, in order to obtain an improvement of healthcare; 2) correct developing strategies, based on scientific evidence, leading to production of recommendations validated by external review; 3) adequate implementation and diffusion in local settings; 4) application with sensible and appropriate clinical discretion. Benefits obtained with the correct use of efficient pediatric guidelines, can be identified at different levels: patients' care (outcome improvement, increased patients' consciousness, influence on public healthcare policy); healthcare professionals (improving quality of clinical decisions, agreement on clinical and therapeutic strategies; medicolegal protection, representing a reference for prospective and retrospective audits); healthcare systems (standardising care, improving efficacy of care; optimising costs). Personal experience in systematic development of emergency pediatric guidelines, applied in a second level Emergency Department is also presented. PMID- 12754455 TI - [The newborn with cardiological problems. The dilemma between malformative and non-malformative pathology]. AB - Cardiovascular impairment is frequent during the neonatal period and can be expression of malformative or not-malformative pathology. In both conditions the clinical presentation is often dramatic with cyanosis and/or heart failure. The neonatologist has to make differential diagnosis as soon as possible, because cardiac malformations in the neonatal period are usually ductus arteriosus dependent and can worsen suddenly after its closure. Since colour Doppler echocardiography is not available in all the neonatal units in order to be helped in the diagnosis, it is very important that neonatologists learn to use the indications obtained with a careful physical examination and with some simple instrumental tests, as chest X-ray, electrocardiogram and blood gas analysis. In this article a review is made of the most frequent heart malformations associated with cyanosis and/or heart failure during the neonatal period (complete transposition of the great arteries, Fallot's tetralogy, tricuspid and pulmonary atresia, aortic coarctation, interventricular septal defect, persistence of ductus arteriosus) and the most common neonatal pathologic conditions simulating congenital heart diseases (persistence of fetal circulation, neonatal transitory myocardial ischemia, hypervolemia, hypoglycemia, hypocalcemia). Some clinical, instrumental and laboratory findings that could be useful for the diagnosis in absence of echocardiography are also reported. PMID- 12754457 TI - Epidemiology of atopy in 220 children. Diagnostic reliability of skin prick tests and total and specific IgE levels. AB - BACKGROUND: We have prospectively studied 220 children attending our Division because they suffered from atopic dermatitis (AD), asthma, and allergic rhinitis (AR), to assess the epidemiology of atopic diseases, and effectiveness of the diagnostic tests commonly used in allergic children. METHODS: Among the 220 children there were 142 males (64.5%) and 78 females (35.5%) aged as follows: 57 (25.9%) 0-2 year-old, 48 (21.8%) 2-4 year-old, 49 (22.3%) 4-6 year-old, 66 (30%) >6 year-old. The diagnosis included family and personal history, physical examination, skin prick tests (SPT) and total and specific IgE (sIgE) levels. We tested inhalant and food allergens. RESULTS: There were 101 asthmatic, 88 with AD, and 31 children with AR. The analysis of variance confirmed the age influence of PRIST with a high significance (p=0.0001). SPTs were prevalent in all groups for Der p, but casein only in 1 group, and Lolium perenne only in 2 groups. RAST showed a higher uniformity, that is CM (cow's milk) and egg for one group, Der p and Lolium perenne for the remaining groups Several correlations among diagnostic tests and the age of children were evaluated with the analysis of variance. CONCLUSIONS: We emphasize that atopic diseases are genetically transmitted, that AD develops at a younger age than asthma (p=0.0052), and that SPTs have a greater effectiveness for inhalant allergens, positive at all age levels; in food allergy (FA) SPTs are less adequate and feasible. PMID- 12754458 TI - [Ultrasound monitoring of the orthopaedic treatment of congenital hip dysplasia carried out within the first 6 months of life]. AB - BACKGROUND: The use of ultrasound scanning in the early diagnosis of congenital hip dysplasia in infants within the first six months of life has become part of the routine in preventive medicine examinations. Ultrasound scanning makes it possible to monitor the development of the neonatal dysplasic hip at any moment and it also tells us whether or not it is responding to treatment and, if it is not, to modify the treatment in order to achieve cure. METHODS: The present study considers pathological hips studied echographically over 12 years in order to assess to what extent ultrasound controls during treatment have influenced the choices and modifications of treatment. The study employed different echographic equipment always with 5 or 7.5 MHz linear probes; the evaluation technique employed was that proposed by Graf. Echography evaluation was always proceded by clinical assessment and followed at the end of treatment by X-ray control of the pelvis. RESULTS: Of 17,938 hip cases assessed echographically from 1 January 1989 to 31 December 2000 (59.5% female and 40.5% male, average age 3 months 17 days) 1534 hips were pathological (IIC, IID, IIIA, IIIB); the prevalence of the condition in females and on the left-hand side was confirmed. In 35 cases (2.19%) the orthopaedic treatment approach undertaken at the beginning was changed on the basis of periodic scans. The normalisation of the alpha angle was achieved on average in 51 days of treatment with the harness; with a minimum time of 30 days in IIC type hips in patients below the age of three months and a maximum time of 98 days in hips of type IIIA in patients aged more than 120 days. CONCLUSIONS: The simplicity and ease of use of ultrasound scanning, its cheapness, rapidity, harmlessness and validity not to mention its sensitivity and specificity make it a reliable, safe instrumental examination that is indispensable in planning the treatment of this pathology. PMID- 12754459 TI - [Respiratory diseases in infants hospitalized during the 1st 2 years of life for viral lower respiratory tract infections: a one-year follow-up]. AB - BACKGROUND: Epidemiologic data suggest strong links between hospitalisation with bronchiolitis in infancy and subsequent higher risk of developing lower respiratory tract infections (LRTI) and/or hyperreactive airway diseases. The aim of this study was to evaluate in an Italian population the natural history of respiratory diseases in children hospitalised for LRTI when they were <2 years. METHODS: An observational, perspective, longitudinal study was performed through telephone interviews. Nine pediatric tertiary care centres participated to the study evaluating a population of 187 children, hospitalised in the previous year (November 1999-April 2000) for bronchiolitis or pneumonia when they were <2 years of age and participated to a previous study on the prevalence of infant LRTI in Italy (RADAR). RESULTS: Twenty-three (12.3%) children had a gestational age <36 weeks. In the 12 months following the first hospitalisation, an elevated frequency of respiratory symptoms was found. Indeed, 152 (81.3%) children suffered from not-requiring-hospital-admission respiratory infections and 21 (11.2%) were hospitalized again for LRTI: 11.6% had bronchiolitis, 23.5% bronchitis and 35.2% pneumonia. In addition, 1.2% had gs;3 infectious episodes and 21.4% gs;6: 68 (36.4%) showed wheezy bronchitis and 17 (9.1%) were reported to have asthma; 132 children (71%) took antibiotics during the last year, 19.4% >3 times; 111 (59.4%) bronchodilators and 49 (26.2%) oral corticosteroids. One year after the first hospitalisation, 19 subjects (10.2%) were found to be positive to at least one class of allergens by prick test or RAST. CONCLUSIONS: Thus, the demonstration of a high morbidity rate for LRTI, wheezing and asthma in this study group during the first year follow-up after hospital admission further support the need for prophylactic interventions to reduce the morbidity and severity of sequelae of LRTI, in particularly in premature children and/or with additional risk factors. PMID- 12754460 TI - [Suicidal behaviour in young people. An epidemiological study in the Verbano Cusio-Ossola Province (years 1988-2000)]. AB - BACKGROUND: The aim of the present study is to investigate the suicide and attempted suicide phenomenon among young people (<25 years old) in the Verbano Cusio-Ossola province from January 1988 to December 2000. METHODS: This epidemiological-descriptive survey is based on the acquisition of data through the examination of model 45 registered at the Verbania Public Prosecutor's office. The data obtained were analysed with SPSS 8.0 software for Windows. The significance of the differences between the rates observed in our group and those observed in Italy in the same period was estimated by calculating SMR and SIR (Standardized Mortality Rates and Standardized Incidence Rates respectively). RESULTS: In the period considered in our study, 13 suicides and 62 attempted suicides were notified to the Court, with a rate of 2.55 and 12.18 per 100,000 inhabitants, respectively. The analysis of SMR and SIR points out that the incidence of suicide and attempted suicide among young people is higher in this province than in Italy. The most frequently used methods to commit suicide are hanging and carbon monoxide poisoning, while drug intoxication prevails in attempted suicide. The most common reasons are disagreements, followed by mental illness, psychosocial factors, loss of a relative and toxic dependence. CONCLUSIONS: The present study means to provide a description of suicide behaviour among young people in a geographic and cultural context, in order to point out its problems and to provide useful information for the diagnosis and prevention. PMID- 12754461 TI - [Complex family association in autoimmune polyendocrine syndrome]. AB - Autoimmune polyglandular syndromes (APS) are conditions characterized by the association of two or more endocrine and non-endocrine autoimmune disorders. Diabetes mellitus type 1 (T1DM) is one of the most frequent components of APS and is often its first symptom. The frequency of autoimmune pathologies in patients affected by T1DM is proportional to the persistance of ICA. Even in first relatives of these patients, an increase in incidence of latent or manifest autoimmune pathology is noticed. The association of T1DM with autoimmune thyroiditis and celiac disease in a girl from a family affected by high incidence of autoimmune pathology is described. The role of gluten in the pathogenesis of T1DM and some other autoimmune conditions in genetically predisposed subjects. Infact studies are still inadequate for demostrating how a gluten-free diet could delay or mitigate the course of T1DM and of other autoimmune pathologies in genetically predisposed subjects. Nevertheless, it is suggested that gluten could represent a starting or a maintenance factor of autoimmune processes and the risk of autoimmune pathologies is proportional to the duration of the exposure to gluten. A screening for a quick singling out of autoimmune pathologies is suggested for T1DM patients, their first relatives and for subjects affected by other autoimmune diseases or celiac disease. PMID- 12754462 TI - Dermatitis herpetiformis in a 30-month-old child. AB - Dermatitis herpetiformis is commonly observed in adults and adolescents. We report one of the youngest cases seen so far-a 30-month-old boy. After clinical investigations an asymptomatic gluten-sensitive enteropathy was diagnosed. Skin lesions resolved with gluten-free diet. Dermatitis herpetiformis should be considered in differential diagnosis of chronic dermatitis in early childhood as well. Monitoring for prevention of complications is the greatest problem at this age. PMID- 12754463 TI - [Cystic dysplasia of rete testis associated with ipsilateral renal agenesis. Case report]. AB - Cystic dysplasia of the rete testis is a rare abnormality often associated with the ipsilateral agenesis of kidney. This malformation is due to a development defect of the mesonephric duct which is the cause of both the dilation of the testicular rete testis and renal agenesis. A case of this rare malformation, showing all the peculiarities described in the medical literature, is presented. A 3 years-4 months boy was examined for an asymptomatic left scrotal mass; thus, he underwent ultrasonography, which showed a multiple tubular and cystic dilatation of left rete testis, associated with the absence of left kidney, afterward confirmed by MAG3-radionuclide scan. Diagnosis was also validated by testicular biopsy. No surgery was required. The child is nowadays under observation and at 2-years follow-up he doesn't show any symptom. According to many authors, a conservative treatment of this benign congenital abnormality is suggested as well as serial ultrasonography to monitor the growth of the testicular mass, which in a longest follow-up, could require surgery. Malignant transformation nor infertility have never been described. PMID- 12754464 TI - Multiple Myeloma 9th International Workshop. May 23-27, 2003, Salamanca, Spain. Abstracts. PMID- 12754494 TI - The C-terminal domain of apolipoprotein A-I contains a lipid-sensitive conformational trigger. AB - Exchangeable apolipoproteins can convert between lipid-free and lipid-associated states. The C-terminal domain of human apolipoprotein A-I (apoA-I) plays a role in both lipid binding and self-association. Site-directed spin-label electron paramagnetic resonance spectroscopy was used to examine the structure of the apoA I C terminus in lipid-free and lipid-associated states. Nitroxide spin-labels positioned at defined locations throughout the C terminus were used to define discrete secondary structural elements. Magnetic interactions between probes localized at positions 163, 217 and 226 in singly and doubly labeled apoA-I gave inter- and intramolecular distance information, providing a basis for mapping apoA-I tertiary and quaternary structure. Spectra of apoA-I in reconstituted HDL revealed a lipid-induced transition of defined random coils and beta-strands into alpha-helices. This conformational switch is analogous to triggered events in viral fusion proteins and may serve as a means to overcome the energy barriers of lipid sequestration, a critical step in cholesterol efflux and HDL assembly. PMID- 12754495 TI - Structural basis for orthogonal tRNA specificities of tyrosyl-tRNA synthetases for genetic code expansion. AB - The archaeal/eukaryotic tyrosyl-tRNA synthetase (TyrRS)-tRNA(Tyr) pairs do not cross-react with their bacterial counterparts. This 'orthogonal' condition is essential for using the archaeal pair to expand the bacterial genetic code. In this study, the structure of the Methanococcus jannaschii TyrRS-tRNA(Tyr)-L tyrosine complex, solved at a resolution of 1.95 A, reveals that this archaeal TyrRS strictly recognizes the C1-G72 base pair, whereas the bacterial TyrRS recognizes the G1-C72 in a different manner using different residues. These diverse tRNA recognition modes form the basis for the orthogonality. The common tRNA(Tyr) identity determinants (the discriminator, A73 and the anticodon residues) are also recognized in manners different from those of the bacterial TyrRS. Based on this finding, we created a mutant TyrRS that aminoacylates the amber suppressor tRNA with C34 65 times more efficiently than does the wild-type enzyme. PMID- 12754496 TI - Amyloid-like filaments and water-filled nanotubes formed by SOD1 mutant proteins linked to familial ALS. AB - Mutations in the SOD1 gene cause the autosomal dominant, neurodegenerative disorder familial amyotrophic lateral sclerosis (FALS). In spinal cord neurons of human FALS patients and in transgenic mice expressing these mutant proteins, aggregates containing FALS SOD1 are observed. Accumulation of SOD1 aggregates is believed to interfere with axonal transport, protein degradation and anti apoptotic functions of the neuronal cellular machinery. Here we show that metal deficient, pathogenic SOD1 mutant proteins crystallize in three different crystal forms, all of which reveal higher-order assemblies of aligned beta-sheets. Amyloid-like filaments and water-filled nanotubes arise through extensive interactions between loop and beta-barrel elements of neighboring mutant SOD1 molecules. In all cases, non-native conformational changes permit a gain of interaction between dimers that leads to higher-order arrays. Normal beta-sheet containing proteins avoid such self-association by preventing their edge strands from making intermolecular interactions. Loss of this protection through conformational rearrangement in the metal-deficient enzyme could be a toxic property common to mutants of SOD1 linked to FALS. PMID- 12754497 TI - Chip and solution detection of DNA hybridization using a luminescent zwitterionic polythiophene derivative. AB - Electronic polymers in aqueous media may offer bioelectronic detection of biospecific interactions. Here we report a fluorometric DNA hybridization detection method based on non-covalent coupling of DNA to a water-soluble zwitterionic polythiophene derivative. Introduction of a single-stranded oligonucleotide will induce a planar polymer and aggregation of the polymer chains, detected as a decrease of the intensity and a red-shift of the fluorescence. On addition of a complementary oligonucleotide, the intensity of the emitted light is increased and blue-shifted. The detection limit of this method is at present approximately 10(-11) moles. The method is highly sequence specific, and a single-nucleotide mismatch can be detected within five minutes without using any denaturation steps. The interaction with DNA and the optical phenomena persists when the polymer is deposited and patterned on a surface. This offers a novel way to create DNA chips without using covalent attachment of the receptor or labelling of the analyte. PMID- 12754498 TI - Pinpoint and bulk electrochemical reduction of insulating silicon dioxide to silicon. AB - Silicon dioxide (SiO(2)) is conventionally reduced to silicon by carbothermal reduction, in which the oxygen is removed by a heterogeneous-homogeneous reaction sequence at approximately 1,700 degrees C. Here we report pinpoint and bulk electrochemical methods for removing oxygen from solid SiO(2) in a molten CaCl(2) electrolyte at 850 degrees C. This approach involves a 'contacting electrode', in which a metal wire supplies electrons to a selected region of the insulating SiO(2). Bulk reduction of SiO(2) is possible by increasing the number of contacting points. The same method was also demonstrated with molten LiCl-KCl CaCl(2) at 500 degrees C. The novelty and relative simplicity of this method might lead to new processes in silicon semiconductor technology, as well as in high-purity silicon production. The methodology may be applicable to electrochemical processing of a wide variety of insulating materials, provided that the electrolyte dissolves the appropriate constituent ion(s) of the material. PMID- 12754499 TI - Direct measurement of triaxial strain fields around ferroelectric domains using X ray microdiffraction. AB - Ferroelectric materials, such as BaTiO(3), have piezoelectric properties that make them attractive for microelectronic and sensing applications. It is well known that the application of mechanical stress or electric field can alter the domain structure in ferroelectrics. Indeed, the constitutive behaviour of a ferroelectric is largely governed by the formation, movement and interaction of its domains. Therefore, it is crucial that the micromechanics of domains and their effect on internal stresses in ferroelectrics be understood. Here we show that the emerging technique of scanning X-ray microdiffraction can be used to measure directly, for the first time, the local triaxial strain fields around 90 degrees domains in single-crystal BaTiO(3). Specifically, residual strain maps in a region surrounding an isolated, approximately 40 microm wide, 90 degrees domain were obtained with 3 microm resolution, revealing significant residual strains. This information is critical for accurate micromechanical modelling of domain behaviour in ferroelectrics. PMID- 12754500 TI - A stable quasi-solid-state dye-sensitized solar cell with an amphiphilic ruthenium sensitizer and polymer gel electrolyte. AB - Dye-sensitized nanocrystalline solar cells (DSC) have received considerable attention as a cost-effective alternative to conventional solar cells. One of the main factors that has hampered widespread practical use of DSC is the poor thermostability encountered so far with these devices. Here we show a DSC with unprecedented stable performance under both thermal stress and soaking with light, matching the durability criteria applied to silicon solar cells for outdoor applications. The cell uses the amphiphilic ruthenium sensitizer cis RuLL'(SCN)(2) (L = 4,4'-dicarboxylic acid-2,2'-bipyridine, L' = 4,4'-dinonyl-2,2' bipyridine) in conjunction with a quasi-solid-state polymer gel electrolyte, reaching an efficiency of >6% in full sunlight (air mass 1.5, 100 mW cm(-2)). A convenient and versatile new route is reported for the synthesis of the heteroleptic ruthenium complex, which plays a key role in achieving the high temperature stability. Ultramicroelectrode voltammetric measurements show that the triiodide/iodide couple can perform charge transport freely in the polymer gel. The cell sustained heating for 1,000 h at 80 degrees C, maintaining 94% of its initial performance. The device also showed excellent stability under light soaking at 55 degrees C for 1,000 h in a solar simulator (100 mW cm(-2)) equipped with a ultraviolet filter. The present findings should foster widespread practical application of dye-sensitized solar cells. PMID- 12754501 TI - Inhibition of hypothalamic carnitine palmitoyltransferase-1 decreases food intake and glucose production. AB - The enzyme carnitine palmitoyltransferase-1 (CPT1) regulates long-chain fatty acid (LCFA) entry into mitochondria, where the LCFAs undergo beta-oxidation. To investigate the mechanism(s) by which central metabolism of lipids can modulate energy balance, we selectively reduced lipid oxidation in the hypothalamus. We decreased the activity of CPT1 by administering to rats a ribozyme-containing plasmid designed specifically to decrease the expression of this enzyme or by infusing pharmacological inhibitors of its activity into the third cerebral ventricle. Either genetic or biochemical inhibition of hypothalamic CPT1 activity was sufficient to substantially diminish food intake and endogenous glucose production. These results indicated that changes in the rate of lipid oxidation in selective hypothalamic neurons signaled nutrient availability to the hypothalamus, which in turn modulated the exogenous and endogenous inputs of nutrients into the circulation. PMID- 12754502 TI - Leuprorelin rescues polyglutamine-dependent phenotypes in a transgenic mouse model of spinal and bulbar muscular atrophy. AB - Spinal and bulbar muscular atrophy (SBMA) is an adult-onset motor neuron disease that affects males. It is caused by the expansion of a polyglutamine (polyQ) tract in androgen receptors. Female carriers are usually asymptomatic. No specific treatment has been established. Our transgenic mouse model carrying a full-length human androgen receptor with expanded polyQ has considerable gender related motor impairment. This phenotype was abrogated by castration, which prevented nuclear translocation of mutant androgen receptors. We examined the effect of androgen-blockade drugs on our mouse model. Leuprorelin, a lutenizing hormone-releasing hormone (LHRH) agonist that reduces testosterone release from the testis, rescued motor dysfunction and nuclear accumulation of mutant androgen receptors in male transgenic mice. Moreover, leuprorelin treatment reversed the behavioral and histopathological phenotypes that were once caused by transient increases in serum testosterone. Flutamide, an androgen antagonist promoting nuclear translocation of androgen receptors, yielded no therapeutic effect. Leuprorelin thus seems to be a promising candidate for the treatment of SBMA. PMID- 12754503 TI - Dynamic imaging of collagen and its modulation in tumors in vivo using second harmonic generation. AB - The content and structure of collagen is essential in governing the delivery of therapeutic molecules in tumors. Thus, simple histological staining of tumor tissue biopsies for collagen could be used to assess the accessibility of molecular therapeutics in tumors. Here we show that it is possible to optically image fibrillar collagen in tumors growing in mice using second-harmonic generation (SHG). Using this noninvasive technique, we estimated relative diffusive hindrance, quantified the dynamics of collagen modification after pharmacologic intervention and provided mechanistic insight into improved diffusive transport induced by the hormone relaxin. This technology could offer basic scientists and clinicians an enhanced ability to estimate the relative penetrabilities of molecular therapeutics. PMID- 12754504 TI - Long-term follow-up studies confirm the stability of the latent reservoir for HIV 1 in resting CD4+ T cells. AB - Latent HIV-1 persists in resting memory CD4+ T cells, even in patients receiving highly active antiretroviral therapy (HAART). It has been unclear how stable this latent reservoir is and whether its persistence reflects replenishment by low level viremia. Here we show that even in treated patients who have had no detectable viremia for as long as 7 years, the reservoir decays so slowly (t(1/2) = 44 months) that eradication is unlikely. PMID- 12754505 TI - SOCS3 negatively regulates IL-6 signaling in vivo. AB - Members of the suppressor of cytokine signaling (SOCS) family are potentially key physiological negative regulators of interleukin-6 (IL-6) signaling. To examine whether SOCS3 is involved in regulating this signaling, we have used conditional gene targeting to generate mice lacking Socs3 in the liver or in macrophages. We show that Socs3 deficiency results in prolonged activation of signal transducer and activator of transcription 1 (STAT1) and STAT3 after IL-6 stimulation but normal activation of STAT1 after stimulation with interferon-gamma (IFN-gamma). Conversely, IL-6-induced STAT activation is normal in Socs1-deficient cells, whereas STAT1 activation induced by IFN-gamma is prolonged. Microarray analysis shows that the pattern of gene expression induced by IL-6 in Socs3-deficient livers mimics that induced by IFN-gamma. Our data indicate that SOCS3 and SOCS1 have reciprocal functions in IL-6 and IFN-gamma regulation and imply that SOCS3 has a role in preventing IFN-gamma-like responses in cells stimulated by IL-6. PMID- 12754506 TI - SOCS3 regulates the plasticity of gp130 signaling. AB - Suppressor of cytokine signaling (SOCS) proteins are feedback inhibitors of the Janus kinase (JAK) and signal transducer and activator of transcription (STAT) signaling pathway. SOCS3 is upregulated by several signals in macrophages and has been implicated as a regulator of various signaling pathways. Here we show that phosphorylation of STAT3 is prolonged in mouse Socs3-deficient macrophages after stimulation with interleukin-6 (IL-6) but not IL-10, indicating that SOCS3 specifically affects signaling mediated by IL-6 and gp130. IL-6 induces a wider transcriptional response in Socs3-deficient macrophages than in wild-type cells; this response is dominated by interferon (IFN)-regulated genes owing to an excess of STAT1 phosphorylation. Thus, SOCS3 functions to control the quality of the response to IL-6 and prevents the activation of an IFN-induced program of gene expression. PMID- 12754507 TI - IL-6 induces an anti-inflammatory response in the absence of SOCS3 in macrophages. AB - Whereas interleukin-6 (IL-6) is a proinflammatory cytokine, IL-10 is an anti inflammatory cytokine. Although signal transducer and activator of transcription 3 (STAT3) is essential for the function of both IL-6 and IL-10, it is unclear how these two cytokines have such opposing functions. Here we show that suppressor of cytokine signaling 3 (SOCS3) is a key regulator of the divergent action of these two cytokines. In macrophages lacking the Socs3 gene or carrying a mutation of the SOCS3-binding site in gp130, the lipopolysaccharide-induced production of tumor necrosis factor (TNF) and IL-12 is suppressed by both IL-10 and IL-6. SOCS3 specifically prevents activation of STAT3 by IL-6 but not IL-10. Taken together, these data indicate that SOCS3 selectively blocks signaling by IL-6, thereby preventing its ability to inhibit LPS signaling. PMID- 12754509 TI - Adaptation shapes patterns of genome evolution on sexual and asexual chromosomes in Drosophila. AB - What advantage might sexual recombination confer? Population genetics theory predicts that asexual genomes are less efficient at eliminating deleterious mutations and incorporating beneficial alleles. Here, I compare patterns of genome evolution in a 40-kb gene-rich region on homologous neo-sex chromosomes of Drosophila miranda. Genes on the non-recombining neo-Y show various signs of degeneration, including transposable-element insertions, frameshift mutations and a higher rate of amino-acid substitution. In contrast, loci on the recombining neo-X show intact open reading frames and generally low rates of amino-acid substitution. One exceptional gene on the neo-X shows evidence for adaptive protein evolution, affecting patterns of variability at neighboring regions along the chromosome. These findings illustrate the limits to natural selection in an asexual genome. Deleterious mutations, including repetitive DNA, accumulate on a non-recombining chromosome, whereas rapid protein evolution due to positive selection is confined to the recombining homolog. PMID- 12754508 TI - Hypotrichosis simplex of the scalp is associated with nonsense mutations in CDSN encoding corneodesmosin. AB - We have identified nonsense mutations in the gene CDSN (encoding corneodesmosin) in three families suffering from hypotrichosis simplex of the scalp (HSS; OMIM 146520). CDSN, a glycoprotein expressed in the epidermis and inner root sheath (IRS) of hair follicles, is a keratinocyte adhesion molecule. Truncated CDSN aggregates were detected in the superficial dermis and at the periphery of hair follicles. Our findings suggest that CDSN is important in normal scalp hair physiology. PMID- 12754510 TI - Positional cloning of a quantitative trait locus on chromosome 13q14 that influences immunoglobulin E levels and asthma. AB - Atopic or immunoglobulin E (IgE)-mediated diseases include the common disorders of asthma, atopic dermatitis and allergic rhinitis. Chromosome 13q14 shows consistent linkage to atopy and the total serum IgE concentration. We previously identified association between total serum IgE levels and a novel 13q14 microsatellite (USAT24G1; ref. 7) and have now localized the underlying quantitative-trait locus (QTL) in a comprehensive single-nucleotide polymorphism (SNP) map. We found replicated association to IgE levels that was attributed to several alleles in a single gene, PHF11. We also found association with these variants to severe clinical asthma. The gene product (PHF11) contains two PHD zinc fingers and probably regulates transcription. Distinctive splice variants were expressed in immune tissues and cells. PMID- 12754511 TI - Gene expression phenotypic models that predict the activity of oncogenic pathways. AB - High-density DNA microarrays measure expression of large numbers of genes in one assay. The ability to find underlying structure in complex gene expression data sets and rigorously test association of that structure with biological conditions is essential to developing multi-faceted views of the gene activity that defines cellular phenotype. We sought to connect features of gene expression data with biological hypotheses by integrating 'metagene' patterns from DNA microarray experiments in the characterization and prediction of oncogenic phenotypes. We applied these techniques to the analysis of regulatory pathways controlled by the genes HRAS (Harvey rat sarcoma viral oncogene homolog), MYC (myelocytomatosis viral oncogene homolog) and E2F1, E2F2 and E2F3 (encoding E2F transcription factors 1, 2 and 3, respectively). The phenotypic models accurately predict the activity of these pathways in the context of normal cell proliferation. Moreover, the metagene models trained with gene expression patterns evoked by ectopic production of Myc or Ras proteins in primary tissue culture cells properly predict the activity of in vivo tumor models that result from deregulation of the MYC or HRAS pathways. We conclude that these gene expression phenotypes have the potential to characterize the complex genetic alterations that typify the neoplastic state, whether in vitro or in vivo, in a way that truly reflects the complexity of the regulatory pathways that are affected. PMID- 12754512 TI - A statistical explanation of visual space. AB - The subjective visual space perceived by humans does not reflect a simple transformation of objective physical space; rather, perceived space has an idiosyncratic relationship with the real world. To date, there is no consensus about either the genesis of perceived visual space or the implications of its peculiar characteristics for visually guided behavior. Here we used laser range scanning to measure the actual distances from the image plane of all unoccluded points in a series of natural scenes. We then asked whether the differences between real and apparent distances could be explained by the statistical relationship of scene geometry and the observer. We were able to predict perceived distances in a variety of circumstances from the probability distribution of physical distances. This finding lends support to the idea that the characteristics of human visual space are determined probabilistically. PMID- 12754513 TI - AKAP150 signaling complex promotes suppression of the M-current by muscarinic agonists. AB - M-type (KCNQ2/3) potassium channels are suppressed by activation of G(q/11) coupled receptors, thereby increasing neuronal excitability. We show here that rat KCNQ2 can bind directly to the multivalent A-kinase-anchoring protein AKAP150. Peptides that block AKAP150 binding to the KCNQ2 channel complex antagonize the muscarinic inhibition of the currents. A mutant form of AKAP150, AKAP(DeltaA), which is unable to bind protein kinase C (PKC), also attenuates the agonist-induced current suppression. Analysis of recombinant KCNQ2 channels suggests that targeting of PKC through association with AKAP150 is important for the inhibition. Phosphorylation of KCNQ2 channels was increased by muscarinic stimulation; this was prevented either by coexpression with AKAP(DeltaA) or pretreatment with PKC inhibitors that compete with diacylglycerol. These inhibitors also reduced muscarinic inhibition of M-current. Our data indicate that AKAP150-bound PKC participates in receptor-induced inhibition of the M current. PMID- 12754515 TI - Experience-dependent slow-wave sleep development. AB - Sleep enhances plasticity in neocortex, and thereby improves sensory learning. Here we show that sleep itself undergoes changes as a consequence of waking experience during a late critical period in cats and mice. Dark-rearing produced a robust and reversible decrement of slow-wave electrical activity during sleep that was restricted to visual cortex and impaired by gene-targeted reduction of NMDA receptor function. PMID- 12754514 TI - The protocadherin Flamingo is required for axon target selection in the Drosophila visual system. AB - Photoreceptor neurons (R cells) in the Drosophila visual system elaborate a precise map of visual space in the brain. The eye contains some 750 identical modules called ommatidia, each containing eight photoreceptor cells (R1-R8). Cells R1-R6 synapse in the lamina; R7 and R8 extend through the lamina and terminate in the underlying medulla. In a screen for visual behavior mutants, we identified alleles of flamingo (fmi) that disrupt the precise maps elaborated by these neurons. These mutant R1-R6 neurons select spatially inappropriate targets in the lamina. During target selection, Flamingo protein is dynamically expressed in R1-R6 growth cones. Loss of fmi function in R cells also disrupts the local pattern of synaptic terminals in the medulla, and Flamingo is transiently expressed in R8 axons as they enter the target region. We propose that Flamingo mediated interactions between R-cell growth cones within the target field regulate target selection. PMID- 12754516 TI - Development of neural mechanisms for reading. AB - The complexities of pediatric brain imaging have precluded studies that trace the neural development of cognitive skills acquired during childhood. Using a task that isolates reading-related brain activity and minimizes confounding performance effects, we carried out a cross-sectional functional magnetic resonance imaging (fMRI) study using subjects whose ages ranged from 6 to 22 years. We found that learning to read is associated with two patterns of change in brain activity: increased activity in left-hemisphere middle temporal and inferior frontal gyri and decreased activity in right inferotemporal cortical areas. Activity in the left-posterior superior temporal sulcus of the youngest readers was associated with the maturation of their phonological processing abilities. These findings inform current reading models and provide strong support for Orton's 1925 theory of reading development. PMID- 12754517 TI - Perceiving numbers causes spatial shifts of attention. AB - Number symbols are part of our everyday visual world. Here we show that merely looking at numbers causes a shift in covert attention to the left or right side, depending upon the number's magnitude. This observation implies obligatory activation of number meaning and signals a tight coupling of internal and external representations of space. PMID- 12754518 TI - Activity-dependent activation of presynaptic protein kinase C mediates post tetanic potentiation. AB - Vesicle exocytosis is mediated by the complex interaction between synaptic vesicle and plasma membrane proteins, many of which are substrates for protein kinases. Exogenous protein kinase activators increase release probability at several mammalian CNS synapses, but the physiological conditions under which presynaptic protein kinases become activated are not known. We report here that calcium/phospholipid-dependent protein kinase C (PKC) is activated by high frequency stimulation and mediates post-tetanic potentiation (PTP) in the rat hippocampus. PMID- 12754519 TI - Identification and quantification of N-linked glycoproteins using hydrazide chemistry, stable isotope labeling and mass spectrometry. AB - Quantitative proteome profiling using stable isotope protein tagging and automated tandem mass spectrometry (MS/MS) is an emerging technology with great potential for the functional analysis of biological systems and for the detection of clinical diagnostic or prognostic marker proteins. Owing to the enormous complexity of proteomes, their comprehensive analysis is an as-yet-unresolved technical challenge. However, biologically or clinically important information can be obtained if specific, information-rich protein classes, or sub-proteomes, are isolated and analyzed. Glycosylation is the most common post-translational modification. Here we describe a method for the selective isolation, identification and quantification of peptides that contain N-linked carbohydrates. It is based on the conjugation of glycoproteins to a solid support using hydrazide chemistry, stable isotope labeling of glycopeptides and the specific release of formerly N-linked glycosylated peptides via peptide- N glycosidase F (PNGase F). The recovered peptides are then identified and quantified by MS/MS. We applied the approach to the analysis of plasma membrane proteins and proteins contained in human blood serum. PMID- 12754520 TI - Turnover-based in vitro selection and evolution of biocatalysts from a fully synthetic antibody library. AB - This report describes the selection of highly efficient antibody catalysts by combining chemical selection from a synthetic library with directed in vitro protein evolution. Evolution started from a naive antibody library displayed on phage made from fully synthetic, antibody-encoding genes (the Human Combinatorial Antibody Library; HuCAL-scFv). HuCAL-scFv was screened by direct selection for catalytic antibodies exhibiting phosphatase turnover. The substrate used was an aryl phosphate, which is spontaneously transformed into an electrophilic trapping reagent after cleavage. Chemical selection identified an efficient biocatalyst that then served as a template for error-prone PCR (epPCR) to generate randomized repertoires that were subjected to further selection cycles. The resulting superior catalysts displayed cumulative mutations throughout the protein sequence; the ten-fold improvement of their catalytic proficiencies (>10(10) M( 1)) resulted from increased kcat values, thus demonstrating direct selection for turnover. The strategy described here makes the search for new catalysts independent of the immune system and the antibody framework. PMID- 12754521 TI - Lectin affinity capture, isotope-coded tagging and mass spectrometry to identify N-linked glycoproteins. AB - We describe here a strategy for the large-scale identification of N-glycosylated proteins from a complex biological sample. The approach, termed isotope-coded glycosylation-site-specific tagging (IGOT), is based on the lectin column mediated affinity capture of a set of glycopeptides generated by tryptic digestion of protein mixtures, followed by peptide-N-glycosidase-mediated incorporation of a stable isotope tag, 18O, specifically into the N-glycosylation site. The 18O-tagged peptides are then identified by multi-dimensional liquid chromatography-mass spectrometry (LC-MS)-based technology. The application of this method to the characterization of N-linked high-mannose and/or hybrid-type glycoproteins from an extract of Caenorhabditis elegans proteins allowed the identification of 250 glycoproteins, including 83 putative transmembrane proteins, with the simultaneous determination of 400 unique N-glycosylation sites. Because the method is applicable to the systematic identification of a wide range of glycoproteins, it should facilitate basic glycobiology research and may be useful for diagnostic applications, such as genome-wide screening for disease-related glycoproteins. PMID- 12754522 TI - Production of knockout rats using ENU mutagenesis and a yeast-based screening assay. AB - The rat is a widely used model in biomedical research and is often the preferred rodent model in many areas of physiological and pathobiological research. Although many genetic tools are available for the rat, methods to produce gene disrupted knockout rats are greatly needed. In this study, we developed protocols for creating N-ethyl-N-nitrosourea (ENU)-induced germline mutations in several rat strains. F1 preweanling pups from mutagenized Sprague Dawley (SD) male rats were then screened for functional mutations in Brca1 and Brca2 using a yeast gap repair, ADE2-reporter truncation assay. We produced knockout rats for each of these two breast cancer suppressor genes. PMID- 12754523 TI - Expression profiling reveals off-target gene regulation by RNAi. AB - RNA interference is thought to require near-identity between the small interfering RNA (siRNA) and its cognate mRNA. Here, we used gene expression profiling to characterize the specificity of gene silencing by siRNAs in cultured human cells. Transcript profiles revealed siRNA-specific rather than target specific signatures, including direct silencing of nontargeted genes containing as few as eleven contiguous nucleotides of identity to the siRNA. These results demonstrate that siRNAs may cross-react with targets of limited sequence similarity. PMID- 12754524 TI - Segregation of object and background motion in the retina. AB - An important task in vision is to detect objects moving within a stationary scene. During normal viewing this is complicated by the presence of eye movements that continually scan the image across the retina, even during fixation. To detect moving objects, the brain must distinguish local motion within the scene from the global retinal image drift due to fixational eye movements. We have found that this process begins in the retina: a subset of retinal ganglion cells responds to motion in the receptive field centre, but only if the wider surround moves with a different trajectory. This selectivity for differential motion is independent of direction, and can be explained by a model of retinal circuitry that invokes pooling over nonlinear interneurons. The suppression by global image motion is probably mediated by polyaxonal, wide-field amacrine cells with transient responses. We show how a population of ganglion cells selective for differential motion can rapidly flag moving objects, and even segregate multiple moving objects. PMID- 12754525 TI - Insulin-regulated hepatic gluconeogenesis through FOXO1-PGC-1alpha interaction. AB - Hepatic gluconeogenesis is absolutely required for survival during prolonged fasting or starvation, but is inappropriately activated in diabetes mellitus. Glucocorticoids and glucagon have strong gluconeogenic actions on the liver. In contrast, insulin suppresses hepatic gluconeogenesis. Two components known to have important physiological roles in this process are the forkhead transcription factor FOXO1 (also known as FKHR) and peroxisome proliferative activated receptor gamma co-activator 1 (PGC-1alpha; also known as PPARGC1), a transcriptional co activator; whether and how these factors collaborate has not been clear. Using wild-type and mutant alleles of FOXO1, here we show that PGC-1alpha binds and co activates FOXO1 in a manner inhibited by Akt-mediated phosphorylation. Furthermore, FOXO1 function is required for the robust activation of gluconeogenic gene expression in hepatic cells and in mouse liver by PGC-1alpha. Insulin suppresses gluconeogenesis stimulated by PGC-1alpha but co-expression of a mutant allele of FOXO1 insensitive to insulin completely reverses this suppression in hepatocytes or transgenic mice. We conclude that FOXO1 and PGC 1alpha interact in the execution of a programme of powerful, insulin-regulated gluconeogenesis. PMID- 12754526 TI - Facing the severe acute respiratory syndrome epidemic. Hope for the best and prepare for the worst. PMID- 12754527 TI - Sample size determination. Influencing factors and calculation strategies for survey research. AB - The paper reviews both the influencing factors and calculation strategies of sample size determination for survey research. It indicate the factors that affect the sample size determination procedure and explains how. It also provides calculation methods (including formulas) that can be applied directly and easily to estimate the sample size needed in most popular situations. PMID- 12754528 TI - Febrile neutropenia. Etiology of infection, empirical treatment and prophylaxis. AB - Much has changed in the treatment of patients with fever and neutropenia, including the patterns of microbial flora and drug resistance, and the drugs used. Gram-positive organisms have overshadowed the gram-negative ones as causes of bacteremia. Changes in therapy may include antimicrobials directed against gram-positive bacteria, resistant gram-negative bacteria, or fungi. Due to the high risk for colonization by vancomycin resistant Enterococci, vancomycin use is restricted as first line empiric therapy unless the patient is at high-risk for serious gram-positive infection. Prophylactic antibiotic therapy may increase the selection of resistant strains and should be avoided. Therapy with colony stimulating factor is only considered for patients who remain severely neutropenic and have documented infections that do not respond to appropriate antibacterial therapy. Patients stratification for risk of infection-associated morbidity and mortality is essential to facilitate treatment decision. PMID- 12754529 TI - Assessment of nutritional rickets in Western Saudi Arabia. AB - OBJECTIVE: To explore the role of radiological examination and certain biochemical values in diagnosis and assessing severity of nutritional rickets. METHODS: Cases of symptomatic nutritional rickets (age range between 3-36 months) seen at King Abdul-Aziz University Hospital, Jeddah, Kingdom of Saudi Arabia, during the period 1997-1999 were studied. Clinical examination by the author of the study, determination of calcium (Ca), phosphate (PO4), alkaline phosphates (ALP), hand and wrist x-rays, were performed for all cases. RESULTS: Sixty cases of nutritional rickets were diagnosed within 2 years (incidence of 0.5%), 38.3% of the patients presented with swollen wrist and 28.3% with bowleg. The bone profile at time of diagnosis: Ca=2.33 +/- 0.23, PO4 = 1.47 +/- 0.40 and ALP = 925 +/- 418. Approximately 81.7% of the patients had normal Ca level, 18.3% had low serum PO4 level, 98.3% showed high value of ALP. X-ray studies indicated that, 58.3% of patients had active rickets, 35% had minimal changes, and 6.7% showed healed rickets. Among those having active rickets 20% had low PO4 level, 83% had normal Ca value, and 100% had high ALP. The mean value +/- SD of biochemical values in this group: Ca = 2.34 +/- 0.24, PO4 = 1.45 +/- 0.42, ALP = 1067 +/- 452. The later was significantly higher compared to other groups (P=0.004) but no significant differences were observed between mean values of other parameters. CONCLUSION: Radiological examination and ALP remains essential to confirm clinical diagnosis of rickets and assessment of severity. PMID- 12754530 TI - Effects of thyroid hormones on basal and stimulated gastric acid secretion due to histamine, carbachol and pentagastrin in rats. AB - OBJECTIVE: Thyroid hormones affect gastric acid secretion. As the mechanism of this effect has not been fully known, in this experimental study the isolated gastric acid secretion of hypothyroid and hyperthyroid rats were compared with control group by the administration of different doses of pentagastrin, histamine and carbachol as gastric acid secretion stimulators. METHODS: This study was carried out in Ahwaz University of Medical Sciences, Ahwaz, Iran in the year 2000. Each group were consisted of 8 rats (N-mari) of both sexes with a mean weight of 246 +/- 5 grams. Hypothyroid and hyperthyroid states were induced respectively by adding methimazole (500 mg/liter) for 20 days and thyroxin (500 microg/liter) for 35 days in animals drinking water. After general anesthesia, by intraperitoneal injection of sodium thiopental (50 mg/kg body weight), celiotomy was carried out quickly. The end of esophagus was tied and a silicon tube (2-2.5 mm) was entered into the stomach via duodenum and fasted in pylor region. The stomach was isolated by cutting the esophagus proximal to the tied region and the proximal part of duodenum and put into cold serous solution. After washing the serous and mucus surfaces by serous and mucus solutions, the stomach was transferred immediately to a tissue bath containing warm serous solution (V= 40 ml, T= 37 degrees C). Gastric acid secretion in isolated stomach stimulated by pentagastrin, carbachol and histamine was measured by wash out technique and automatic titrator. Moreover, to study the effect of thyroid hormones on gastric acid secretion a number of dose-dependent experiments after the administration of different doses of histamine (50, 100, 150, 200 micromol), carbachol (50, 100, 150, 200 micromol), and pentagastrin (30, 60, 90, 120 ug/kg body weight) were performed. RESULTS: Both basal and histamine, carbachol, pentagastrin stimulated acid secretion decreased and increased in hypothyroid and hyperthyroid groups compared with control group. CONCLUSION: It seems that thyroid hormones have not exert their effects by changing the cholinergic, gastrin and histamine receptors but probably by alerting the number or size of the secretory cells in stomach. PMID- 12754531 TI - Radioiodine treatment of hyperthyroidism. Success rate and influence of thyrostatic medication. AB - OBJECTIVE: The aim of this study is to evaluate the response rate of hyperthyroidism to radioactive iodine (RAI) treatment, optimum effective dose, effect of pretreatment with thyrostatic medications, etiology, ophthalmopathy, mortality and cancer incidence post RAI treatment. METHODS: Retrospective study analysis of 360 patients records who received RAI treatment (dose 5-15 mCi) for hyperthyroidism in Hamad Medical Corporation, Qatar between 1984-1999, treated and analyzed. Follow-up data was available in 215 patients, with a follow-up range of 2-10 years, of these 84 were males and 131 were females, with an age range of 12-74 years. Eighty percent were toxic diffuse goiter, 13.5% were toxic multinodular goiter and 6.5% were toxic single nodule. Eighty-seven percent had been pre-treated with anti-thyroid medications. Free thyroxine4, and thyroid stimulating hormone were recorded at diagnosis; 6 months, one year and yearly post RAI treatment. RESULTS: The incidence of hypothyroidism was 55.8% at 6 months and 67.9% at one year. There was no significant difference in the response rate to different doses of RAI treatment groups (50-59%, p=0.46). The response rate was significantly higher in the group without pre-treatment with anti thyroid medications (95% versus 80.9%, p<0.0001) and 27.4% of our patients had ophthalmopathy. There was no significant worsening or new development of ophthalmopathy post RAI treatment. Three of our patients developed cancer: one with colonic, one with breast and one with acute leukemia. The mortality rate according to the age group was linear in the positive direction of age and the highest was 74-year-old (10.5 per 10,0000 population). CONCLUSION: Radioactive iodine treatment is an effective modality for definitive treatment of hyperthyroidism with long-term cure approaching 80%. Response rate was not related to gender, etiology or RAI dosage. Pre-treatment with anti-thyroid medication reduces the response rate. Radioactive iodine treatment has no significant influence on ophthalmopathy, mortality or thyroid cancer. PMID- 12754532 TI - Thyroid function and thyroid autoimmunity in patients with type 1 diabetes mellitus. AB - OBJECTIVE: An association between diabetes mellitus and autoimmune thyroid disease is well known. We have investigated the prevalence of thyroid dysfunction and autoimmunity in type 1 diabetic patients. METHODS: Seventy-nine type 1 diabetic patients were recruited in the study, and underwent complete investigations for thyroid function, which included free thyroxine, free tri iodothyronine, and thyroid stimulating hormone, of those only 64 patients had performed thyroid autoantibodies (TAb); anti- thyroid peroxidase antibodies (TPOAb) or antimicrosomal antibodies and thyroglobulin antibodies (TgAb). They were compared with 127 healthy subjects matched for sex and age. This study was carried out at the National Center for Diabetes, Endocrinology and Genetics, Jordan University, Amman, Jordan between 2000 and 2001. RESULTS: In the diabetic group, 7 cases (8.9%) of thyroid dysfunction were detected, 4 of these were diagnosed as subclinical hypothyroidism, whereas the other 3 had overt hypothyroidism and were on thyroxine replacement therapy. In the control group, 6 (4.7%) subjects were diagnosed as subclinical hyperthyroidism. There was a significant difference in thyroid function variables between diabetics and controls. Among type 1 diabetic patients, 7 (9.2%) had thyroid autoantibodies, 5 with positive TPOAb only and 2 with positive TAb; TPOAb or antimicrosomal antibodies and TgAb; compared with 8 (6.3%) in the control group, 4 with positive TPOAb only and 4 with positive TAb; TPOAb or antimicrosomal antibodies and TgAb P=0.68. CONCLUSION: Biochemical thyroid dysfunction and thyroid autoimmunity were evident in type 1 diabetics who were apparently euthyroid, with no significant difference between diabetics and controls. PMID- 12754533 TI - Is hypertension well controlled in hypertensive diabetics. AB - OBJECTIVE: To determine the degree of blood pressure (BP) control in hypertensive diabetics and to study the types of antihypertensive agents used for BP control. METHODS: A cross-sectional study was carried out on hypertensive diabetics followed in the outpatient clinic of King Abdul-Aziz University Hospital, Jeddah, Kingdom of Saudi Arabia from January 2000 to February 2001. Patient's age, sex, duration of diabetes and its control, duration of hypertension and the type of antihypertensive agents used, were noted. Patients were classified according to the degree of systolic and diastolic BP control into 4 groups: systolic group-I (< or = 120 mmHg), group-II (121-130 mmHg), group-III (131-140 mmHg), group-IV (>140 mmHg); diastolic group-I (< or = 80 mmHg), group-II (81-85 mmHg), group-III (86-90 mmHg), group-IV (>90 mmHg). RESULTS: A total of 230 patients were included with a mean age of 61 years and an equal male to female ratio. Mean duration of diabetes was 14 years and 9 years for hypertension. Five of 230 (2.2%) were in systolic group-I, 28/230 (12%) in group-II, 94/230 (41%) in group-III, 103/230 (44.8%) in group-IV; while 7/230 (3%) were in diastolic group-I, 30/230 (13%) in group-II, 92/230 (40%) in group-III, and 101/230 (43.9%) in group-IV. Angiotensin converting enzyme-inhibitors were used in 163/230 (70.9%) followed by diuretics in 99/230 (43%), Calcium channel-blockers in 62/230 (27%), and B-blockers in 25/230 (10.9%). CONCLUSION: Only a small percentage of hypertensive diabetics met the recommended BP for diabetics. Efforts should be made by both patients and physician to achieve better BP control. PMID- 12754534 TI - Door to needle time in administering thrombolytic therapy for acute myocardial infarction. AB - OBJECTIVE: Thrombolytic therapy is a standard treatment for patients presenting with acute myocardial infarction (MI). Early administration of these agents is crucial for the outcome of management. This audit was conducted to evaluate the time between arrival to emergency department (ED) and the administration of thrombolysis (door to needle time). METHODS: Data was collected from patients admitted to the Coronary Care Unit of Riyadh Medical Complex (RMC), Riyadh, Kingdom of Saudi Arabia, a 1500-bed community hospital, with a diagnosis of acute MI and received thrombolytic therapy over a one-year period (April 1999 to April 2000). The time between arrival to the ED to the time of administration of thrombolytic therapy was obtained as well as the time of onset of chest pain up to presentation to the hospital, and the outcome (all cause mortality) post treatment. RESULTS: A total of 271 patients (256 males) admitted to RMC with a diagnosis of acute MI received thrombolytic therapy over a one-year duration. The median door to needle time was 95 minutes. The median time of onset of chest pain to arrival to ED was 5 hours (300 minutes). The outcome of these patients obtained either alive was 260 (96%) or dead was 11 (4%) (P < 0.00001). CONCLUSION: The door to needle time was relatively similar to other centers. The delay in administering thrombolytic therapy should be reduced to a target of <70 minutes from onset of symptoms. Delay in presentation to the hospital was more important and factors contributing to this delay should be looked for and corrected. Another audit is needed to evaluate the implementation of these recommendations. PMID- 12754535 TI - Ureteric injuries during obstetric and gynecologic procedures. AB - OBJECTIVE: To review the ureteric injuries resulting from obstetric and gynecologic procedures with a comparative analysis of current pertinent literature. METHODS: From January 1994 -December 1997, the medical records of all patients who sustained ureteric injuries as a result of obstetric or gynecologic procedures and managed at the Princess Basma Teaching Hospital, North of Jordan were reviewed. The clinical presentations, investigations, features of injury and treatment modalities were studied. RESULTS: There were 17 patients with 19 ureteric injuries incurred during obstetric or gynecologic procedures during the study period. Patients were relatively young and presented with loin pain. The left lower ureter was injured in 11 cases. Hysterectomy, alone, accounted for 13 injuries mainly in association with malignancy. Deliveries, in general, were associated with very low rate of injury. Ureteric ligation was the most common mechanism of injury (47%). Injuries were intraoperatively recognized in 41.2% (7/17) of cases. Patients were treated by either endourological or formal surgical repair. The overall success rate after an average of 32.3 months of follow-up was achieved in 89.5% (17/19). This outcome was not significantly altered by either the features of injury or by the treatment schedules. CONCLUSION: Iatrogenic ureteric injury is still a major cause of harm and concern. The time taken to detect the injury remains the most important morbidity related factor. Recent trends towards earlier intervention and the use of various endourological means of repair deserve support and promotion. PMID- 12754536 TI - Maternal mortality in a teaching hospital in Sudan. AB - OBJECTIVE: Although there is a great advances in obstetrics in recent years, a hard core of maternal mortality still lingers and is considered to be an irreducible minimum. Nevertheless there has been worldwide decline in maternal mortality over the past decades. Yet maternal mortality is still the major public health problem throughout the world. METHODS: A retrospective study carried out at the Medani Teaching Hospital, Medani City, Sudan. Case notes were reviewed for all the 877 women died during the period 1985-1999. All parturients were followed for at least 6 weeks after delivery so that deaths due to sequel of delivery and puerperium were also included. The data was analyzed and the literature was reviewed to compare the results of similar studies. RESULTS: Out of the total number of deliveries in that period (N=44,605), 877 women died with a rate of 1,966 per 100,000. This rate showed a decline each year, being 2,661 per 100,000 during the period 1985-1989. The mortality rate during 1990-1994 was 2,021 per 100,000 and 1,363 per 100,000 during the last period of 1995-1999. Sepsis was the cause of death in almost one third of cases, while malaria accounted for 37.2%. The number of preventable deaths had decreased steadily. CONCLUSION: The study highlighted the existence of a serious and preventable obstetrical problem. Sepsis, malaria hemorrhage and hypertensive disorders are the main contributory factors. Poor antenatal care, poor intranatal care, and poor provision of health services add more to the problem. Research into the problem of malaria, sterilization, anesthesia, vascular accidents, and indication for cesarean section together with social and medical improvements will definitely reduce the maternal morbidity and mortality. PMID- 12754537 TI - Routine ultrasound in acute retention of urine. AB - OBJECTIVE: To evaluate the frequency of urological abnormalities in routine urinary tract ultrasonography (renal and pelvic) in patients with urinary retention secondary to benign prostatic hyperplasia. METHODS: All patients presented to Salmaniya Medical Complex, Bahrain with acute retention of urine secondary to benign prostatic hyperplasia (BPH) in the period between January 2001 and December 2001 were included. The frequency of urological abnormalities, other than BPH, was obtained. RESULTS: One hundred patients were enrolled with a mean age of 67 years. Forty-one patients (41%) had other urological abnormalities. Among these, 3 cases of malignancy were discovered incidentally. A case of renal cell carcinoma, which was completely excised, and 4 cases of bladder tumor, 2 were new cases and 2 were previously known cases of cancer bladder. Other urological abnormalities were renal stones (9 cases), renal cysts (9 cases), hydronephrosis (14 cases) and bladder stones (5 cases). Asymptomatic non-urological abnormalities were gallstones (3 cases), liver cirrhosis (one case) and hepatic hemangioma (one case). Renal impairment was found in 18% of all patients and 80% with hydronephrosis. Four patients had hypoechoic nodules, and all had cancer prostate. CONCLUSION: Significant fraction of patients with acute urinary retention due to BPH have another pathology; although the majority are trivial and it did not influence the immediate management, some are life threatening such as renal cell carcinoma and bladder tumor. Hydronephrosis can be missed if one depends solely on renal biochemistry. Thus, routine evaluation of such patients with pelvic and renal ultrasonography is justified. PMID- 12754538 TI - Plasma and red blood cells membrane lipid concentration of sickle cell disease patients. AB - OBJECTIVE: The present study aimed to determine the concentration of plasma and red blood cells (RBCs) membrane lipids in Saudi sickle cell disease (SCD) patients. METHODS: This study was carried out at the Hematology Clinic, King Abdul-Aziz University Hospital, Jeddah, Kingdom of Saudi Arabia from October 1998 to October 1999. Lipid concentrations were determined in plasma and RBC membrane of 81 SCD patients and 66 normal healthy matched individuals (control). Different lipid parameters were measured according to standardized enzymatic assay methods. RESULTS: The plasma concentrations of total cholesterol and low density lipoprotein cholesterol of SCD patients were significantly decreased (p<0.001), whereas the plasma concentrations of high density lipoprotein phospholipids were significantly increased (p<0.001). The plasma concentrations of apo A and apo B were significantly decreased (p<0.001) in SCD patients. However, the concentration of total cholesterol of RBC membrane was significantly increased (p<0.001) in SCD patients, while the phospholipid content was significantly decreased (p<0.001). CONCLUSION: The significant increase of RBC membrane cholesterol concentration in SCD patients possibly is responsible to the change in RBC membrane fluidity that may play a direct role in the sickling phenomenon of RBCs in SCD. PMID- 12754539 TI - Causes of death in the Eastern Mediterranean Region during the years 1998-2000. AB - OBJECTIVE: This paper aims to consider the available mortality data as an insight to epitomize the mortality pattern and the main leading causes of death specific to the Easter Mediterranean Region (EMR) in order to set priorities for future research in the region. METHODS: Data was taken from the last 3 World Health Organization (WHO) mortality statistics. Crude mortality rates were compared between the 6 WHO regions for the years 1998, 1999 and 2000. Proportional mortality and standardized proportional mortality ratios were calculated for the causes of death and types of malignant neoplasm deaths and compared between the EMR and the other regions of the world. Spearman coefficient rank-order correlation was calculated to detect significant correlation between the ranking of the main causes of death in EMR and the 3 basic demographic, socio-economic and health care indicators in the 6 regions of the world. RESULTS: The results of this study showed that approximately 9 per 1,000 of the world's population die annually. Africa and Europe have the highest mortality rates. The main causes of death worldwide are cardiovascular diseases, infectious or parasitic diseases, malignant neoplasm, infectious respiratory diseases and other respiratory diseases. In EMR, approximately 8 per 1,000 die annually. The causes of death in EMR can be classified into 3 categories. 1) Non-prominent in EMR as compared to other regions of the world. 2) Prominent in EMR and significantly correlated to the basic indicators. 3) Prominent in EMR but not related to the basic indicators. These include deaths due to wars, congenital anomalies, perinatal conditions, genitourinary diseases, endocrine disorders, road traffic accidents, cancer bladder, lymphoma leukemia CONCLUSION: The results of this study emerged the need for extensive epidemiological studies to investigate thoroughly the main causes of death influencing mortality in EMR, specially that they coincide with the health consequences of depleted uranium. Also, most of these health conditions were previously described among the United States and European veterans who served in the Gulf War. PMID- 12754541 TI - Profile of child and adolescent psychiatry in Oman. AB - OBJECTIVE: To find the common behavioral problems in youth, their co-morbidity, treatment, and other variables in Oman. METHODS: All patients who attended the child psychiatric clinic in Sultan Qaboos University Hospital (SQUH), Muscat, (the only child psychiatry clinic in Sultanate of Oman) for a 3-month period were investigated for behavioral problems, particularly hyperactivity (by Conners' Questionnaire). The diagnosis was based on the Diagnostic and Statistical Manual of Mental Disorders, 4th edition. RESULTS: Of the total 212 cases, Omani nationals constituted 89% and the rest were expatriates. Sixty five percent were boys, 57% were of school age, 26% were adolescents, 9% were preschoolers and 5% were toddlers. Consanguinity among their parents was high at 52%. The majority (60%) of the patients exhibited hyperactivity. Aggression was found in 49%, while stealing and lying were found in 25% and 22%. Approximately one-third suffered from headache and abdominal pain. The majority (62%) were of below-normal intelligence. Anxiety affected 14%, depression 4%, and conduct disorders 8%. One third of the patients were taking psychostimulants and another one-third were taking tricyclics. The results were compared with those from other developing countries. CONCLUSION: Behavioral problems among children and adolescents are high in Oman, same as other developing countries, though the type of disorders is different here. This calls for the attention of the health policy makers for diagnosing and treating such disorders in Oman as well as in the other developing countries. PMID- 12754540 TI - The management of acute severe asthma in a pediatric intensive care unit. AB - OBJECTIVE: The hospitalization and mortality rates incurred from acute childhood asthma continue to rise in the past decade. The purpose of this study is to examine the outcome, morbidity and the management of children admitted with acute asthma to our pediatric intensive care unit (PICU) and compare it with those described in the literature. METHODS: Medical records of all children admitted with acute severe asthma to PICU at King Khalid University Hospital, Riyadh, Kingdom of Saudi Arabia over an 8-year period (1994-2001) were reviewed. RESULTS: Fifty-six patients were analyzed. The male to female ratio was 1.3:1 and the mean age was 3.6 +/- 2.8 years. The mean duration of symptoms prior to admission was 2 +/- 1.5 days with 39.3% <24 hours. A positive family history of allergy was present in two third of patients. The average stay in PICU was 2 +/- 0.9 days. Seventy-three percent of patients received prophylaxis bronchodilator therapy before hospital admission including inhaled steroid in 62%. All the patients received nebulized salbutamol and intravenous corticosteroid. Two third of our patients received nebulized ipratropium bromide and 62% intravenous aminophylline. From arterial blood gases analysis, 46.4% had hypercapnia (PaCO2 >45 mmHg). None of our patients required mechanical ventilation. Only 2 patients developed pneumomediastinum with pneumothorax that has resolved spontaneously without intervention. There were no deaths among our 56 patients admitted to PICU. CONCLUSION: We conclude that the mortality and morbidity in children with severe asthma, who require PICU admissions are minimal, provided optimal early use of bronchodilators and intravenous steroids. Using this approach, it could also be possible to avoid mechanical ventilation and shorten the duration of hospital admission. PMID- 12754542 TI - Rheumatoid arthritis. Radiological changes in the cervical spine. AB - OBJECTIVE: To describe the radiographic cervical spine changes in rheumatoid arthritis patients. METHODS: Forty-nine patients (37 females and 12 males) diagnosed with rheumatoid arthritis at King Khalid University Hospital, Riyadh, Kingdom of Saudi Arabia, between June 1998 and June 2000, were studied for their radiographic cervical spine changes. Their mean age at disease onset was 41.4 +/- 13.4 years (range of 18-73) and mean duration of the disease was 9.1 +/- 6.28 years (range of 2-34). Their demographic data including rheumatoid factor status was obtained. Standard conventional radiographs of cervical spine were obtained to study the cervical spine changes. RESULTS: Cervical spine radiographic changes were found in 34 patients (27 females and 7 males), 10 had subluxation (7 with atlanto-axial subluxation, 2 with sub-axial subluxation, and one with lateral subluxation). No vertical impaction was seen. Erosion of odontoid process was seen in one patient. All were rheumatoid seropositive. CONCLUSION: Cervical spine changes in patients with rheumatoid arthritis are common, in particular subluxation in the upper cervical spine. Our study showed somewhat lesser prevalence of these changes. These were clinically correlated with disease duration, female sex, and rheumatoid factor, but were not statistically significant. PMID- 12754543 TI - Fever of unknown origin. AB - This report describes a 3-year-old female child with prolonged fever in which a very extensive work-up resulted in the discovery of the underlying congenital anomaly causing her symptom. PMID- 12754544 TI - Shigella flexneri bacteremia in a child. AB - We report an unusual and lethal case of Shigella flexneri septicemia in an 8-year old Saudi handicapped child from a social home presenting with severe toxic megacolon and acute abdomen secondary to fulminant necrotizing enterocolitis. PMID- 12754545 TI - Isolation of Salmonella paratyphi A from a patient with nephrolithiases. AB - We describe a case of Salmonella paratyphi A isolated from urine of a 37-year-old Saudi patient who is a known case of nephrolithiasis and hydronephrosis with frequent admission for management of renal stones. History of enteric fever was not documented and urinary schistosomiasis in such a patient from endemic area is a strong possibility. Relevant literature was discussed. PMID- 12754546 TI - Isolated agenesis of the gallbladder. AB - We report a rare case of agenesis of the gallbladder, which was misdiagnosed as cholecystitis. This is the first reported case from the Middle East. Despite advances in biliary imaging, the diagnosis is usually made at surgery. Like most patients, our patient became asymptomatic after the surgery. Extensive dissection to exclude the presence of gallbladder in an ectopic site is discouraged. PMID- 12754547 TI - Pancreatic pseudocyst. AB - Gallstones cause more than two-thirds of the acute pancreatitis episodes in the Kingdom of Saudi Arabia (KSA). The majority of these attacks are often mild and self-limiting. However, some are associated with complications; most common of which is development of pancreatic pseudocyst. This complication was reported to be rare in some areas of KSA. The author reports his personal experience with pancreatic pseudocysts that he encountered over 9 years of surgical practice in one of the busiest hospitals in the Eastern Province of KSA and discusses various management options. PMID- 12754548 TI - Nasal polyps masking a unilateral choanal atresia. PMID- 12754549 TI - Gallium-67 lymph node localization in toxic Kikuchi-Fujimoto's disease. PMID- 12754551 TI - Induction of labor with vaginal prostaglandin-E2 in women with one previous cesarean section. PMID- 12754550 TI - Sandhoff disease in an extreme preterm baby with bilateral syndactyly. PMID- 12754552 TI - The association of acute myocardial infarction and pregnancy loss in young female with primary antiphospholipid syndrome. PMID- 12754553 TI - Diclofenac suppositories in the treatment of bone and joint diseases. A forgotten route. PMID- 12754554 TI - Trend of antibiotic resistance in 1316 Shigella strains isolated in Bahrain. PMID- 12754555 TI - Etiology of chronic diarrhea. PMID- 12754556 TI - Sleep and child mental health. PMID- 12754557 TI - Electrocardiographic abnormalities in neurological diseases. AB - OBJECTIVE: To evaluate electrocardiographic abnormalities in patients with neurologic diseases. METHODS: We studied 161 patients with neurologic disorders by analyzing the 12-lead electrocardiogram during the pathological process. An expert who did not know anything about the patients evaluated the traces. RESULTS: Neurological process included brain tumor (41%), stroke (27.3%), cerebral aneurysm (15.5%), subarachnoid hemorrhage (6.8%), subdural hemorrhage (5%), and head injury (4.4%). Electrocardiograms were normal in 61% of cases, and the most frequent abnormality was ventricular repolarization (23.7%). The presence of T waves (4.6%) and prolonged QT intervals (8.8%) was the most characteristic of brain injuries. CONCLUSION: We observed a lower incidence of electrocardiographic abnormalities than that described in the literature. PMID- 12754558 TI - Percutaneous transluminal septal alcoholization for the treatment of refractory hypertrophic obstructive cardiomyopathy: initial experience in the Federal District. AB - OBJECTIVE: To analyze the efficacy of percutaneous transluminal septal alcoholization in the treatment of refractory obstructive hypertrophic cardiomyopathy (HOC). METHODS: The patients were referred for alcoholization after Doppler echocardiography. Before and after alcoholization, the intraventricular pressure gradient was recorded. Alcoholization was performed with a 3mL injection of absolute alcohol through a coronary angioplasty balloon catheter. The procedure was concluded after a significant reduction or abolition of the pressure gradient. RESULTS: Of 22 patients, 18 (81.8%) successfully concluded the procedure with a reduction in intraventricular pressure gradient at baseline (from 67.6+/-24.2 mmHg to 3.8+/-1.9 mmHg, p<0.005) and after extrasystole (from 110.4+/-24.2 mmHg to 9.6+/-2.6 mm Hg, p<0.005). A significant reduction in mean interventricular septal thickness (from 2+/-0.3 mm to 1.7+/-0.2 mm, p<0.005) and in peak pressure gradient (from 90.7+/-23.5 mmHg to 6.1+/-1.4 mmHg, p<0.005) was observed on Doppler echocardiography after 6 months, when all patients were in functional class I. The most frequent acute complication, present in 11% of the patients, was the need for definitive pacing implantation. Relapse of the symptoms and reappearance of the pressure gradient occurred in 16.6% of the patients. One patient (5.5%) died probably due to a diffuse coronary spasm prior to the procedure, and another died suddenly on late follow-up. CONCLUSION: Percutaneous transluminal septal alcoholization is effective and safe in the treatment of HOC. PMID- 12754559 TI - Risk Factors, biochemical markers, and genetic polymorphisms in early coronary artery disease. AB - OBJECTIVE: To assess the risk factors, lipid and apolipoprotein profile, hemostasis variables, and polymorphisms of the apolipoprotein AI-CIII gene in early coronary artery disease (CAD). METHODS: Case-control study with 112 patients in each group controlled by sex and age. After clinical evaluation and nutritional instruction, blood samples were collected for biochemical assays and genetic study. RESULTS: Familial history of early CAD (64 vs 39%), arterial hypertension (69 vs 36%), diabetes mellitus (25 vs 3%), and previous smoking (71 vs 46%) were more prevalent in the case group (p<0.001). Hypertension and diabetes were independent risk factors. Early CAD was characterized by higher serum levels of total cholesterol (235 +/-6 vs 209 +/- 4 mg/dL), of LDL-c (154 +/ 5 vs 135 +/- 4 mg/dL), triglycerides (205 +/- 12 vs 143 +/- 9 mg/dL), and apolipoprotein B (129 +/- 3 vs 105 +/- 3 mg/dL), and lower serum levels of HDL-c (40 +/- 1 vs 46 +/- 1 mg/dL) and apolipoprotein AI (134 +/- 2 vs 146 +/- 2mg/dL) [p<0.01], in addition to an elevation in fibrinogen and D-dimer (p<0.02). The simultaneous presence of the rare alleles of the APO AI-CIII genes in early CAD are associated with hypertriglyceridemia (p=0.03). CONCLUSION: Of the classical risk factors, hypertension and diabetes mellitus were independently associated with early CAD. In addition to an unfavorable lipid profile, an increase in the thrombotic risk was identified in this population. An additive effect of the APO AI-CIII genes was observed in triglyceride levels. PMID- 12754560 TI - Influence of angiotensin II receptor subtypes of the paraventricular nucleus on the physiological responses induced by angiotensin II injection into the medial septal area. AB - OBJECTIVE: We determined the effects of losartan and PD 123319 (antagonists of the AT1 and AT2 angiotensin receptors, respectively), and [Sar1, Ala8] ANG II (a relatively peptide antagonist of angiotensin receptors) injected into the paraventricular nucleus (PVN) on water and 3% NaCl intake, and the diuretic, natriuretic, and pressor effects induced by administration of angiotensin II (ANG II) into the medial septal area (MSA) of conscious rats. METHODS: Holtzman rats were used. Animals were anesthetized with tribromoethanol (20 mg) per 100 grams of body weight, ip. A stainless steel guide cannula was implanted into the MSA and PVN. All drugs were injected in 0.5-microl volumes for 10-15 seconds. Seven days after brain surgery, water and 3% NaCl intake, urine and sodium excretion, and arterial blood pressure were measured. RESULTS: Losartan (40 nmol) and [Sar1, Ala8] ANG II (40 nmol) completely eliminated whereas PD 123319 (40 nmol) partially blocked the increase in water and sodium intake and the increase in arterial blood pressure induced by ANG II (10 nmol) injected into the MSA. The PVN administration of PD 123319 and [Sar1, Ala8] ANG II blocked whereas losartan attenuated the diuresis and natriuresis induced by MSA administration of ANG II. CONCLUSION: MSA involvement with PVN on water and sodium homeostasis and arterial pressure modulation utilizing ANGII receptors is suggested. PMID- 12754562 TI - Infective endocarditis. Surgical therapy. AB - OBJECTIVE: To assess the major causes of surgical morbidity and mortality in patients with infective endocarditis operated upon in a regional cardiology center. METHODS: Thirty-four patients underwent surgical treatment for infective endocarditis. Their ages ranged from 20 to 68 years (mean of 40.6) and 79% were males. Their NYHA functional classes were as follows: IV - 19 (55.8%) patients; III - 12 (35.2%) patients; II - 3 (8.8%) patients. Blood cultures were positive in only 32% of the cases. Eight patients had already undergone previous cardiac surgery, whose major indication (82.3%) was heart failure refractory to clinical treatment. RESULTS: Four (11.7%) patients died at the hospital. Follow-up was complete in 26 (86%) patients. Five (14.7%) patients died later, 12, 36, 48, 60, and 89 months after hospital discharge. Of the 21 patients being currently followed up, 1 is in NYHA functional class III, and 5 in NYHA functional class II. CONCLUSION: A high degree of clinical suspicion, at an early diagnosis, and indication of surgical treatment prior to deterioration of left ventricular function and installation of generalized sepsis may improve prognosis. PMID- 12754561 TI - Lipid peroxidation and nitric oxide inactivation in postmenopausal women. AB - OBJECTIVE: To assess the effect of endogenous estrogens on the bioavailability of nitric oxide (.NO) and in the formation of lipid peroxidation products in pre- and postmenopausal women. METHODS: NOx and S-nitrosothiols were determined by gaseous phase chemiluminescence, nitrotyrosine was determined by ELISA, COx (cholesterol oxides) by gas chromatography, and cholesteryl linoleate hydroperoxides (CE18:2-OOH), trilinolein (TG18:2-OOH), and phospholipids (PC-OOH) by HPLC in samples of plasma. RESULTS: The concentrations of NOx, nitrotyrosine, COx, CE18:2-OOH, and PC-OOH were higher in the postmenopausal period (33.8+/-22.3 microL; 230+/-130 nM; 55+/-19 ng/microL; 17+/-8.7 nM; 2775+/-460 nM, respectively) as compared with those in the premenopausal period (21.1+/-7.3 microM; 114+/-41 nM; 31+/-13 ng/microL; 6+/-1.4 nM; 1635+/-373 nM). In contrast, the concentration of S-nitrosothiols was lower in the postmenopausal period (91+/ 55 nM) as compared with that in the premenopausal p in the premenopausal period (237+/-197 nM). CONCLUSION: In the postmenopausal period, an increase in nitrotyrosine and a reduction of S-nitrosothiol formation, as well as an increase of COx, CE18:2-OOH and PC-OOH formation occurs. Therefore, NO inactivation and the increase in lipid peroxidation may contribute to endothelial dysfunction and to the greater risk for atherosclerosis in postmenopausal women. PMID- 12754563 TI - Endomyocardial fibrosis in infancy. AB - The patient was a 4-month-old infant, who underwent persistent ductus arteriosus interruption with titanium clips at the age of 13 days and, since the age of 2 months, had crises of hypoxia and hypertonicity. After clinical investigation, the presence of pulmonary hypertension was confirmed and left ventricular inflow tract obstruction was suspected. The patient underwent surgical treatment at the age of 4 months, during which right and left ventricular endocardial fibrosis was identified. The fibrosis was resected, but the infant had an unfavorable clinical evolution with significant diastolic restriction and died on the sixth postoperative day. Anatomicopathological and surgical findings suggested endomyocardial fibrosis, although that pathology is very rare at the patient's age. PMID- 12754564 TI - [Case 2/03 - Female, 73-year-old, with diabetes mellitus and chest pain since 2 days (PUC/Campinas-Faculdade de Ciencias Medicas)]. PMID- 12754565 TI - [Case 4/2003-Instituto do Coracao do Hospital das Clinicas da FMUSP]. PMID- 12754566 TI - [C-reactive protein and cardiovascular disease. Basis of scientific evidence]. PMID- 12754567 TI - [Standardization of techniques and devices for ergometric and ergospirometric exams]. PMID- 12754569 TI - Glucose consume and growth of E. coli under electromagnetic field. AB - E. coli was submitted to a 5G electromagnetic field generated by a alternate 60 Hz voltage source. The differences on growth and glucose consume in control and exposed groups were evaluated using the non-parametric Mann-Whitney U-test. There was a significant difference in glucose consume and growth in E. coli after 8 hours of exposition to electromagnetic field. It can be concluded that electromagnetic field had a positive effect in consume of glucose and growth of E. coli. The cause of these results can be explained by an increasing of glucose entrance through membrane due to the stimulated transport system via Facility Diffusion or cyclotron resonance. The growth can be caused by shortening of lag phase and excitement of log phase. PMID- 12754568 TI - Phenotypic and molecular characterization of Salmonella Enteritidis strains isolated in Sao Paulo, Brazil. AB - In Sao Paulo State, Brazil, the epidemic increase in isolation of Salmonella Enteritidis has been observed since 1994. A total of 105 S. Enteritidis strains (72 from human and 33 from non-human sources) isolated during the period 1975 1995, previously characterized by phage typing, was analyzed by antimicrobial susceptibility, plasmid profile, and ribotyping. Over 70% of the strains were susceptible to all antimicrobial agents tested, however, multiple resistance to antimicrobials was observed among the studied strains, mainly those from hospitalized patients. Phage type 8 (PT-8) was predominant among the strains isolated during the period of 1975-1992, but in the following years, PT-4 was the most frequent phage type identified. Seven different plasmid profiles were detected and 96% of the isolates harbored a plasmid of approximately 36 MDa. Ribotyping discriminated fourteen ribotypes (R1 to R14) among the strains examined. By analysis of dendrogram the strains were included in three groups with similarity level of 60%. The obtained results indicate that, a single ribotype (R11), determined for PT-4 strains isolated from 1993, characterizes the epidemic clone of S. Enteritidis in our region. PMID- 12754571 TI - High prevalence of GB virus C/hepatitis G virus RNA among Brazilian blood donors. AB - The aim of this study was to investigate the presence of the Hepatitis G Virus on a population of blood donors from S o Paulo, Brazil and to evaluate its association to sociodemographic variables. Two RT-PCR systems targeting the putative 5'NCR and NS3 regions were employed and the former has shown a higher sensitivity. The observed prevalence of HGV-RNA on 545 blood donors was 9.7% (CI 95% 7.4;12.5). Statistical analysis depicted an association with race/ethnicity, black and mulatto donors being more frequently infected; and also with years of education, less educated donors presenting higher prevalences. No association was observed with other sociodemographic parameters as age, gender, place of birth and of residence. DNA sequencing of nine randomly chosen isolates demonstrated the presence of genotypes 1, 2 and 3 among our population but clustering of these Brazilian isolates was not detected upon phylogenetic analysis. PMID- 12754570 TI - Opportunistic infections in patients with AIDS admitted to an university hospital of the Southeast of Brazil. AB - Opportunistic diseases in HIV-infected patients have changed since the introduction of highly active anti-retroviral therapy (HAART). This study aims at evaluating the frequency of associated diseases in patients with AIDS admitted to an university hospital of Brazil, before and after HAART. The medical records of 342 HIV-infected patients were reviewed and divided into two groups: group 1 comprised 247 patients before HAART and, group 2, 95 patients after HAART. The male-to-female rate dropped from 5:1 to 2:1for HIV infection. There was an increase in the prevalence of tuberculosis and toxoplasmosis, with a decrease in Kaposi's sarcoma, histoplasmosis and cryptococcosis. A reduction of in-hospital mortality (42.0% vs. 16.9%; p = 0.00002) has also occurred. An agreement between the main clinical diagnoses and autopsy findings was observed in 10 out of 20 cases (50%). Two patients with disseminated schistosomiasis and 2 with paracoccidioidomycosis are reported. Overall, except for cerebral toxoplasmosis, it has been noticed a smaller proportion of opportunistic conditions related to severe immunosuppression in the post HAART group. There was also a significant reduction in the in-hospital mortality, possibly reflecting improvement in the treatment of the HIV infection. PMID- 12754572 TI - Canine visceral leishmaniasis in Barra de Guaratiba, Rio de Janeiro, Brazil: assessment of risk factors. AB - Barra de Guaratiba is a coastal area of the city of Rio de Janeiro where American visceral leishmaniasis (AVL) is endemic. Although control measures including killing of dogs and use of insecticides have been applied at this locality, the canine seroprevalence remains at 25% and during 1995 and 1997 eight autochthonous human cases were notified. In order to evaluate factors related to the increase of the risk for Leishmania (Leishmania) chagasi infection in dogs we have screened 365 dogs by anti-Leishmania immunofluorescent antibody test (IFAT) and captured sandflies in the domestic and peridomestic environment. Some variables related to the infection were assessed by uni- and multivariate analysis. The distance of the residence from the forest border, its altitude and the presence of the opossum Didelphis marsupialis in the backyard, were found predictor factors for L. (L.) chagasi infection in dogs in Barra de Guaratiba. The presence of Lutzomyia longipalpis in the peridomestic environment indicates the possibility of appearence of new human cases. Our data also suggest the presence of a sylvatic enzootic cycle at this locality. PMID- 12754573 TI - Domestic Rhodnius ecuadoriensis (Hemiptera, Reduviidae) infestation in Northern Peru: a comparative trial of detection methods during a six-month follow-up. AB - Two passive methods in the assessment of intradomiciliary infestation by Rhodnius ecuadoriensis were tested: (i) the Gomes Nu ez sensor box (GN), (ii) sheets of white typing paper and (iii) one active timed manual method. The study was carried out in the Alto Chicama River Valley, Province of Gran Chim , Department of La Libertad. The study design consisted of an initial searching of triatomines inside of the domestic environment by the manual capture active procedure (man/hour) covering all the studied houses. Then, matched pairs of GN boxes and paper sheets were simultaneously installed in the bedrooms of 207 households distributed in 19 localities. A comparative prospective trial of these passive detection devices were monitored at 2, 4 and, finally 6 months follow-up. Parasitological Trypanosoma rangeli and/or T. cruzi infections were investigated in two houses with high level of infestation by R. ecuadoriensis. 16.9% of the 207 households investigated by an initial active manual method were infested with R. ecuadoriensis. The proportion of infested houses fluctuated from 6.2 to 55.5% amongst the 19 localities investigated. T. rangeli natural infection was detected in R. ecuadoriensis specimens collected in two households. Parasite rates in the bugs ranged from 16.6 to 21.7% respectively. The most striking fact was an average rate of salivary gland infection ranging from 7.4 to 8.3%. At the end of the sixth month period, a cumulative incidence of 31.4% of positive GN boxes against 15.9% for paper sheets was recorded. All three methods combined detected domestic infestation in 129 (62.3%) of the 207 houses studied in the 19 localities. The range of houses infested varies from 6.7% to 92.9%. In areas with low bug density infestation rates, the methodology experienced in our studies, seems to be the best choice for investigations on domestic R. ecuadoriensis populations. PMID- 12754574 TI - A retrospective study of histopathological findings in 894 cases of megacolon: what is the relationship between megacolon and colonic cancer? AB - Patients with megaesophagus (ME) have increased prevalence of cancer of the esophagus. In contrast, a higher incidence of colorectal cancer is not observed in patients with megacolon (MC). MC is very common in some regions of Brazil, where it is mainly associated with Chagas disease. We reviewed the pathology records of surgical specimens of all patients submitted for surgical resection of MC in the Hospital das Clinicas of the Faculty of Medicine of Ribeirao Preto (HC FMRP), from the University of Sao Paulo. We found that 894 patients were operated from 1952 until 2001 for MC resection. Mucosal ulcers, hyperplasia and chronic inflammation were frequently found, while polyps were uncommon. No patients with MC presented any type of colonic neoplasm. This observation reinforces the hypothesis that MC has a negative association with cancer of the colon. This seems to contradict the traditional concept of carcinogenesis in the colon, since patients with MC presents important chronic constipation that is thought to cause an increase in risk for colon cancer. MC is also associated with other risk factors for cancer of colon, such as hyperplasia, mucosal ulcers and chronic inflammation. In ME these factors lead to a remarkable increase in cancer risk. The study of mucosal cell proliferation in MC may provide new insights and useful information about the role of constipation in colonic carcinogenesis. PMID- 12754575 TI - Scanning electron microscopy of third-instar sarcophagid (Diptera: Sarcophagidae) recovered from a mummified human corpse in Thailand. AB - The third-instar of an unidentified sarcophagid, recovered from a mummified body of a 32-yr-old Thai male was examined using scanning electron microscopy (SEM). Although the morphological features of this larva are similar to the other sarcophagid larvae, some features could be helpful for species identification, which is a basic requirement for estimation of postmortem interval in forensic investigation. These features included number and arrangement of papillae on the anterior spiracle, structure of spines, size of circumspiracular tubercles at caudal segment and branching peculiarity of the posterior spiracular hairs. This information could benefit future identification of the sarcophagid larvae that exist in Thailand. PMID- 12754576 TI - Analysis of genetic relatedness between populations of Aedes aegypti from different geographic regions of Sao Paulo state, Brazil. AB - RAPD markers have been used for the analysis of genetic differentiation of Aedes aegypti, because they allow the study of genetic relationships among populations. The aim of this study was to identify populations in different geographic regions of the S o Paulo State in order to understand the infestation pattern of A. aegypti. The dendrogram constructed with the combined data set of the RAPD patterns showed that the mosquitoes were segregated into two major clusters. Mosquitoes from the Western region of the S o Paulo State constituted one cluster and the other was composed of mosquitoes from a laboratory strain and from a coastal city, where the largest Latin American port is located. These data are in agreement with the report on the infestation in the S o Paulo State. The genetic proximity was greater between mosquitoes whose geographic origin was closer. However, mosquitoes from the coastal city were genetically closer to laboratory reared mosquitoes than to field-collected mosquitoes from the S o Paulo State. The origin of the infestation in this place remains unclear, but certainly it is related to mosquitoes of origins different from those that infested the West and North region of the State in the 80's. PMID- 12754577 TI - A retrospective evaluation of a score system adopted by the Ministry of Health, Brazil in the diagnosis of pulmonary tuberculosis in childhood: a case control study. AB - Based on a retrospective case-control study we evaluated the score system adopted by the Ministry of Health of Brazil (Ministerio da Saude - MS), to diagnose pulmonary tuberculosis (PTB) in childhood. This system is independent of bacteriological or histopathological data to define a very likely (> or = 40 points), possible (30-35 points) or unlikely (< or = 25 points) diagnosis of tuberculosis. Records of hospitalized non-infected HIV children at the Instituto de Puericultura e Pediatria Martagao Gesteira of Federal University of Rio de Janeiro (IPPMG-UFRJ), were reviewed. Patients were adjusted for age and divided in two different groups: 45 subjects in the case group (culture-positive) [mean of age = 10.64 mo; SD 9.66]; and 96 in the control group (culture-negative and clinic criteria that dismissed the disease) [mean of age = 11.79 mo.; SD 11.31]. Among the variables analyzed, the radiological status had the greater impact into the diagnosis (OR = 25.39), followed by exposure to adult with tuberculosis (OR = 10.67), tuberculin skin test >10mm (OR = 8.23). The best cut-off point to the diagnosis of PTB was 30 points, where the score system was more accurate, with sensitivity of 88.9% and specificity of 86.5%. PMID- 12754578 TI - ABO System: molecular mimicry of Ascaris lumbricoides. AB - A. lumbricoides has been associated to the ABO System by various authors. The objective was to detect ABO System epitopes in A. lumbricoides of groups O, A, B and AB patients. 28 adult parasites were obtained from children to be used as assay material. The patients ABO blood groups were determined. Extracts of A. lumbricoides [AE] were prepared by surgical remotion of the cuticle and refrigerated mechanical rupture. Agglutination Inhibition (AI) and Hemoagglutination Kinetics (HK) tests were used with the [AE]. Of the 28 [AE], eight belonged to O group patients, 15 to A group, three to B group and the remaining two to AB children. The AI Test showed A epitopes in two [AE] of group A patients and B epitopes in two [AE] of group B patients. The HK Test showed B antigenic determiners in two [AE] of group B patients and in two [AE] of group AB patients as well as A antigenic determiners in one [AE] of A group patient. Of the 28 [AE] studied in both tests B epitopes were detected in all [AE] from B and AB patients and A epitopes in three of the 15 [AE] of group A patients. The experiments carried out suggest that A. lumbricoides might absorb A and B antigens from the host, and/or modify the cuticular carbohydrates expression as a kind of antigenic mimicry. PMID- 12754579 TI - UNA virus: first report of human infection in Argentina. AB - Una virus (UNAV), Togaviridae family, is widely distributed in South America, where infections have been detected in mosquitoes and vertebrate hosts (humans, birds and horses). We analyzed human sera from Cordoba inhabitants aged 44 to 89 years and using a neutralization test, we found a prevalence of UNAV antibodies of 3.8% (3/79). The low titers detected suggest past infections probably acquired in rural areas of the Province of Cordoba (central Argentina). None sera were found positive for MAYV neutralizing antibodies. This is the first report of human infections by UNAV in Argentina. PMID- 12754580 TI - Tuberculous brain abscess in a patient with AIDS: case report and literature review. AB - Tuberculous brain abscesses in AIDS patients are considered rare with only eight cases reported in the literature. We describe the case of a 34-year-old woman with AIDS and previous toxoplasmic encephalitis who was admitted due to headache and seizures. A brain computed tomography scan disclosed a frontal hypodense lesion with a contrast ring enhancement. Brain abscess was suspected and she underwent a lesion puncture through a trepanation. The material extracted was purulent and the acid-fast smear was markedly positive. Timely medical and surgical approaches allowed a good outcome. Tuberculous abscesses should be considered in the differential diagnosis of focal brain lesions in AIDS patients. Surgical excision or stereotactic aspiration, and antituberculous treatment are the mainstay in the management of these uncommon lesions. PMID- 12754581 TI - Liver abscess due to Salmonella enteritidis in a returned traveler with HIV infection: case report and review of the literature. AB - Bacteremia due to non-typhi Salmonella is more frequent in patients infected with the human immunodeficiency virus (HIV). However, focal complications have been rarely described. We report a case of liver abscess due to Salmonella enteritidis in an HIV-infected patient who recently returned to Sao Paulo, Brazil, from a trip in the Caribbean. A good clinical and radiological response was seen with both percutaneous catheter drainage and antibiotic treatment. To our knowledge, this is the first culture proven case of non-typhi Salmonellaliver abscess in an HIV-infected patient in Brazil. PMID- 12754582 TI - Man and the animals are merely a passage and channel for food: Leonardo da Vinci, 1452-1519. PMID- 12754583 TI - Hematological findings in Noonan syndrome. AB - OBJECTIVE: Noonan syndrome is a multiple congenital anomaly syndrome, and bleeding diathesis is considered part of the clinical findings. The purpose of this study was to determine the frequency of hemostatic abnormalities in a group of Noonan syndrome patients. METHOD: We studied 30 patients with clinical diagnosis of Noonan syndrome regarding their hemostatic status consisting of bleeding time, prothrombin time, activated partial thromboplastin time and thrombin time tests, a platelet count, and a quantitative determination of factor XI. RESULTS: An abnormal laboratory result was observed in 9 patients (30%). Although coagulation-factor deficiencies, especially factor XI deficiency, were the most common hematological findings, we also observed abnormalities of platelet count and function in our screening. CONCLUSIONS: Hemostatic abnormalities are found with some frequency in Noonan syndrome patients (30% in our sample). Therefore, we emphasize the importance of a more extensive hematological investigation in these patients, especially prior to an invasive procedure, which is required with some frequency in this disorder. PMID- 12754584 TI - Effects of continuous versus bolus infusion of enteral nutrition in critical patients. AB - PURPOSE: Enteral alimentation is the preferred modality of support in critical patients who have acceptable digestive function and are unable to eat orally, but the advantages of continuous versus intermittent administration are surrounded by controversy. With the purpose of identifying the benefits and complications of each technique, a prospective controlled study with matched subjects was conducted. PATIENTS AND METHODS: Twenty-eight consecutive candidates for enteral feeding were divided into 2 groups (n = 14 each) that were matched for diagnosis and APACHE II score. A commercial immune-stimulating polymeric diet was administered via nasogastric tube by electronic pump in the proportion of 25 kcal/kg/day, either as a 1-hour bolus every 3 hours (Group I), or continuously for 24 hours (Group II), over a 3-day period. Anthropometrics, biochemical measurements, recording of administered drugs and other therapies, thorax X-ray, measurement of abdominal circumference, monitoring of gastric residue, and clinical and nutritional assessments were performed at least once daily. The principal measured outcomes of this protocol were frequency of abdominal distention and pulmonary aspiration, and efficacy in supplying the desired amount of nutrients. RESULTS: Nearly half of the total population (46.4%) exhibited high gastric residues on at least 1 occasion, but only 1 confirmed episode of pulmonary aspiration occurred (3.6%). Both groups displayed a moderate number of complications, without differences. Food input during the first day was greater in Group II (approximately 20% difference), but by the third day, both groups displayed similarly small deficits in total furnished volume of about 10%, when compared with the prescribed diet. CONCLUSIONS: Both administration modalities permitted practical and effective administration of the diet with frequent registered abnormalities but few clinically significant problems. The two groups were similar in this regard, without statistical differences, probably because of meticulous technique, careful monitoring, strict patient matching, and conservative amounts of diet employed in both situations. Further studies with additional populations, diagnostic groups, and dietetic prescriptions should be performed in order to elucidate the differences between these commonly used feeding modalities. PMID- 12754585 TI - Sexing the human skull through the mastoid process. AB - OBJECTIVE: The purpose of this study was to evaluate the significance for sex determination of the measurement of the area formed by the xerographic projection of 3 craniometric points related to the mastoid process: the porion, asterion, and mastoidale points. METHOD: Sixty skulls, 30 male and 30 female, were analyzed. A xerographic copy of each side of the skull was obtained. On each xerographic copy, the craniometric points were marked to demarcate a triangle. The area (mm ) of the demarcated triangle for each side of the skull (right (D) and left (E) sides) was determined, and the total value of these measures (T) was calculated. RESULTS: Concerning the right area of the male and female skulls, 60% of the values overlapped; for the left area, 51.67% overlapped, and for the total area, 36.67% overlapped. The analysis of the differences between the sexes in the areas studied was significant for the 3 areas. Regarding the total area, which is the preferred measurement because of the asymmetry between the sides of the skull, the value of the mean was 1505.32 mm for male skulls, which was greater than the maximum value obtained in the female skulls. The value of the mean for female skulls was 1221.24 mm , less than the minimum value obtained for the male skulls. CONCLUSIONS: This study demonstrates a significant result in the 3 studied areas, (D), (E), and (T). The total area values show less overlapping of values between the sexes, and therefore can be used for sexing human skulls. For the population studied, values of the total area that were greater than or equal to 1447.40 mm belonged to male crania (95% confidence). Values for this area that were less than or equal to 1260.36 mm belonged to female crania (95% confidence). PMID- 12754586 TI - C cells in the thyroid of pinealectomized rats. AB - PURPOSE: To study quantitatively C cells in the thyroids of non-isogenic rats to determine the possible effects of pinealectomy on the number of these cells, and consequently on the synthesis and secretion of calcitonin. METHODS: Twenty male rats of an outbred strain (200-300 g) were used in the present study. One group of 10 animals was pinealectomized 50 days prior to sacrifice. Thyroid tissue was stained for calcitonin (Dako Corporation) at a 1:1500 dilution. The number of C cells observed was expressed as number of cells/cm . Data were analyzed statistically by Mann-Whitney test. RESULTS: The number of C cells in pinealectomized and normal animals ranged from 489 to 2084 per cm and 227 to 1584 per cm , respectively, a difference that was statistically significant (P <0.05). CONCLUSIONS: These results showed consistent differences in the number of C cells after pinealectomy when compared to controls. We believe that pinealectomy increases the number of C cells in the rat thyroid. PMID- 12754587 TI - Cholestasis in a murine experimental model: lesions include hepatocyte ischemic necrosis. AB - OBJECTIVE: To establish a murine experimental model of bile duct obstruction that would enable controlled observations of the acute and subacute phases of cholestasis. METHODOLOGY: Adult male isogenic BALB/c mice underwent a bile duct ligation (22 animals) or a sham operation (10 animals). Fifteen days after surgery, or immediately after the animal's death, macroscopic findings were noted and histological study of the liver, biliary tree, and pancreas was performed (hematoxylin-eosin and Masson trichromic staining). RESULTS: Beginning 24 hours after surgery, all animals from the bile duct ligation group presented progressive generalized malaise. All animals presented jaundice in the parietal and visceral peritoneum, turgid and enlarged liver, and accentuated dilatation of gallbladder and common bile duct. Microscopic findings included marked dilatation and proliferation of bile ducts with accentuated collagen deposits, frequent areas of ischemic necrosis, hepatic microabscesses, and purulent cholangitis. Animals from the sham operation group presented no alterations. CONCLUSION: We established a murine experimental model of induced cholestasis, which made it possible to study acute and subacute tissue lesions. Our data suggests that in cholestasis, hepatic functional ischemia plays an important role in inducing hepatic lesions, and it also suggests that the infectious process is an important factor in morbidity and mortality. PMID- 12754588 TI - Burkitt-like lymphoma in an infant: a case report. AB - Childhood non-Hodgkin's lymphomas, including Burkitt and Burkitt-like, are rarely diagnosed in infants. A case of B-cell lymphoma in a 13-month-old girl with extensive abdominal disease, ascites, pleural effusion, and tumor lysis syndrome is reported. Phenotypic analysis showed a germinal center B-cell phenotype, and a B-cell clonality was confirmed by polymerase chain reaction. There was no evidence of Epstein-Barr and HIV infection. The case herein reported emphasizes the need for considering the diagnosis of lymphoma even in very young children. PMID- 12754589 TI - Falciform ligament abscess: report of a case. AB - Falciform ligament abscess is rare. We report a case of a 65-year-old man who presented with right upper quadrant abdominal pain, postprandial fullness, and fever. Computed tomography disclosed a cylindrical mass in the anterior abdomen that aroused suspicion of a hepatic abscess. At laparoscopic surgery, an abscess of the falciform ligament was found and drained. Two months later, the patient developed recurrence of the abscess secondary to acute calculous cholecystitis. Abscess drainage and cholecystectomy were performed. The presence of right uppper quadrant abdominal pain, epigastric tenderness, fever, leukocytosis, and a mass in the anterior abdomen should arouse suspicion of falciform ligament abscess. Its treatment consists of abscess drainage. PMID- 12754590 TI - Bronchiolitis, respiratory syncytial virus, and recurrent wheezing: what is the relationship? AB - Various follow-up studies of children hospitalized with bronchiolitis caused by respiratory syncytial virus have demonstrated that a significant proportion of infants (50%) have recurrent wheezing during childhood. Nevertheless, the relationship between these two entities, if any, has not been established. In order to explain this observation, several hypotheses have been proposed. The first suggests that some children could have an individual predisposition to bronchiolitis caused by respiratory syncytial virus and recurrent wheezing. The virus could be a marker of this condition, and the individual predisposition could in turn be related to an individual hypersensitivity to common allergens (atopy), airway hyperreactivity, or to some disorder related to pulmonary anatomy or physiology that was present before the acute episode of bronchiolitis. Another hypothesis proposes that respiratory syncytial virus could be directly responsible for recurrent wheezing. During an episode of bronchiolitis, the damage in the airway mucosa caused by the vital inflammatory response to infection contributes to sensitivity to other allergens or exposes irritant receptors, resulting in recurrent wheezing. For this review, we analyzed the studies that discuss these hypotheses with the purpose of clarifying the mechanisms for the important issue of recurrent wheezing in childhood. PMID- 12754591 TI - Breastfeeding: making the difference in the development, health and nutrition of term and preterm newborns. AB - Breastfeeding is the natural and safe way of feeding small infants, providing nutritional, immunological, psychological and economic recognized and unquestionable advantages. These qualities are especially important in premature infants, because of their vulnerability. Despite highly desirable, there is, in general, little success in breastfeeding preterm infants, especially in special care neonatal units. There are evidences that a high supportive hospital environment, with an interdisciplinary team, makes possible to these infants to be breastfed. In this article, the authors present an up-to-date review about the components of human milk and its unique characteristics, as well as describes aspects that make the breast milk particularly suitable for feeding the premature newborn. PMID- 12754622 TI - The prognostic impact of hormone receptors and c-erbB-2 in pregnancy-associated breast cancer and their correlation with BRCA1 and cell cycle modulators. AB - A population-based series of 122 patients with pregnancy-associated breast carcinomas was histologically revised and the relationship between hormone receptors, c-erbB-2, BRCA1, p27, cyclin E, and cyclin D1 was studied. The 5-year overall survival was 41%; 70% had tumor size >20 mm; 72% had metastasized to regional lymph nodes; 95% were histologic grade II or III; 66% and 75% were negative for estrogen and progesterone receptor, respectively; and c-erbB-2 expression was high (44%). BRCA1 expression was reduced in 33% of the cases. The expression of p27, cyclin D1, and cyclin E was low, 11%, 9%, and 16%, respectively. Cyclin D1 was positively associated with the hormone receptors (p< or =0.01). In multivariate analysis, lymph node status, progesterone receptor, and c-erbB-2 were significant prognostic factors. In subdividing the group according to lymph node status, c-erbB-2 and progesterone receptor retained a prognostic significance in the node positive group only. In conclusion, pregnancy associated breast carcinomas are aggressive tumors, with low expression of hormone receptors, BRCA1, p27, and cyclin E and D1, and high expression of c-erbB 2. PMID- 12754623 TI - An osteoclast-rich tumor of the gastrointestinal tract with features resembling clear cell sarcoma of soft parts: reports of 6 cases of a GIST simulator. AB - Six cases are reported of an osteoclast-rich tumor of the gastrointestinal tract that should be segregated from GIST. Five of the cases were located in the small bowel and one in the stomach. The age of the patients ranged from 13 to 37 years. The tumors behaved aggressively, with metastases to regional lymph nodes, liver, and other intra-abdominal sites. Microscopically, the tumor cells were medium sized, predominantly oval, relatively monomorphic, diffusely immunoreactive for S 100-protein, and negative for CD117, CD34, HMB-45, and Mart-1. They were admixed with scattered osteoclast-like, multinucleated giant cells which were S-100 protein negative and KP1-positive. One case studied cytogenetically had the karyotype 46XX t(12;22)(q13;q12). The cases here reported are interpreted as examples of a distinctive type of gastrointestinal neoplasm which shares some features with clear cell sarcoma of soft parts (melanoma of soft parts), including in one case the chromosomal translocation that is characteristically associated with that entity. PMID- 12754624 TI - Papillary endothelial hyperplasia of the breast: the great impostor for angiosarcoma: a clinicopathologic review of 17 cases. AB - Seventeen cases of papillary endothelial hyperplasia (PEH, Masson's vegetant intravascular hemangioendothelioma) involving breast or mammary subcutaneous tissues are described. The mean patient age was 59; 14 (82%) were female and 12 (71%) presented with a mass. Nine women had mammographic evaluation, 3 of whom had microcalcifications. Five neoplasms were discovered by routine mammography. Sixteen cases were 2.7 cm or less in greatest dimension, and 8 (47%) were associated with a thrombus and/or cavernous hemangioma. Follow-up in 10 cases (up to nearly 8 years) showed no recurrences. Fifty-nine percent of the cases were received at AFIP for consultation with a working diagnosis of angiosarcoma. Features that help distinguish PEH from angiosarcoma include circumscription of the lesion, location in a vessel or association with thrombus, and papillary architecture without significant cytologic atypia or areas of solid growth. The recognition of the morphologic features of this lesion and its inclusion in the differential diagnosis of vascular mammary tumors will reduce the likelihood of its misdiagnosis as an angiosarcoma and avoid unnecessary and aggressive therapy. PMID- 12754625 TI - The concept of bronchioloalveolar cell adenocarcinoma: redefinition, a critique of the 1999 WHO classification, and an ultrastructural analysis of 155 cases. AB - Bronchioloalveolar cell adenocarcinoma (BACA) is bronchioloalveolar because (1) it arises in bronchioles and alveoli and (2) differentiates into bronchiolar and alveolar cells. Every entity possesses unique characteristics that separate it from other entities. The unique characteristic of BACA is its cell type. Lepidic growth is a clue to the cell type and, even though present in the vast majority, is not unique or absolutely essential. Because of the algebraic nature of concepts, the degree of differentiation, the extent of lepidic growth, and the degree of stromal desmoplasia cannot be used as definitional requirements. Likewise, in malignant tumors, absence of stromal invasion cannot be required. An epistemologically valid definition of BACA is proposed and a study of 155 cases defined this way and examined ultrastructurally is presented. PMID- 12754627 TI - Charles Oberling (1895-1960): a herald of modern oncology. PMID- 12754626 TI - Progressive transformation of germinal centers: a clinicopathological study of 42 Japanese patients. AB - To clarify the clinicopathological features of progressive transformation of germinal center (PTGC) unrelated to nodular lymphocyte predominant Hodgkin's lymphoma in Japanese patients, we reviewed 42 cases and compared the results with those of the United States and Germany. Our results were similar to theirs, with male predominance (M/F ratio, 3:1) and the presentation of a solitary asymptomatic enlarged lymph node in the head and neck area as the common features. However, in Japan, PTGC occurs more frequently in elderly patients. In this study, 12 (29%) of the patients with PTGC were aged 60 years or more. Thirteen patients (31%) with lymphadenopathy in the neck and head area had developed localized chronic inflammation (chronic sialoadenitis=4, chronic tonsillitis=3, infectious epidermal cyst=2) or an autoimmune disorder (hyperthyroidism=2 and bronchial asthma=2). None of the patients developed a malignant lymphoma during the follow-up period of 5 to 238 months (median 27 months). Histologically, in a single longitudinal section of the lymph node, the PTGC occupied up to 5% of the total follicles in 22 patients, 5-10% in 10, 10-20% in 7, and more than 20% in 3. In 5 (12%) patients, an association with prominent marginal zone hyperplasia was also noted. This study also indicates that nodal involvement by various low-grade B-cell lymphomas exhibiting marginal zone distribution patterns should be considered as a differential diagnosis of PTGC. Moreover, in Japan, PTGC is thought to be involved in the etiology of florid reactive follicular hyperplasia in elderly patients. PMID- 12754628 TI - Images in pathology. Conversation between needle biopsies. PMID- 12754629 TI - Images in pathology. Pulmonary pathology of severe acute respiratory syndrome (SARS). PMID- 12754630 TI - Images in pathology. Morphologic features of response to Gleevec (Imatinib) treatment of GIST. PMID- 12754632 TI - Images is pathology. World's largest papillary renal carcinoma? PMID- 12754631 TI - Images in pathology. Food for thought! Vegetable cells in histopathological sections. PMID- 12754633 TI - Images in pathology. Prostatic intraepithelial corpora amylacea. PMID- 12754634 TI - Leiomyoma of the urethra: report of 3 cases of a rare entity. AB - Leiomyomas are benign tumors of smooth muscle origin that can occur at any location in the urinary tract. Although rare, they are the most common mesenchymal neoplasms of the urethra. They are more frequent in females than in males. Only 3 cases of leiomyomas of the male urethra and multiple cases in the female have been reported in the English-language literature. We report 2 additional cases of leiomyoma of the male urethra and 1 more case of the female urethra and describe their differential diagnoses and management. PMID- 12754636 TI - Poorly differentiated extraskeletal myxoid chondrosarcoma with t(9;22)(q22;q11) translocation presenting initially as a solid variant devoid of myxoid areas. AB - Extraskeletal myxoid chondrosarcoma (EMC) is a rare soft tissue tumor associated with the translocation t(9;22)(q22;q11-12). Although it has a typical microscopic appearance its morphologic spectrum is wide. We report a case of clinically aggressive, poorly differentiated EMC showing the characteristic translocation, which presented initially as a poorly differentiated sarcoma devoid of myxoid areas in the upper arm of an 85-year-old man. The recurrent tumor contained scattered myxoid areas, which merged imperceptibly with the poorly differentiated areas. Some myxoid areas contained necrotic foci surrounded by viable cells giving rise to a pseudorosette-like arrangement. There were epithelioid foci. This case confirms that solid variants of EMC may exist. Poorly differentiated EMC may have a worse prognosis than classic EMC. PMID- 12754635 TI - Tumor to tumor metastasis: report of two cases and review of the literature. AB - Tumor-to-tumor metastases are uncommon. The most frequent donor tumor is lung cancer, while renal cell carcinoma (RCC) is by far the most common recipient. In this report, a carcinoma of the uterine cervix metastasizing to an RCC and a urothelial carcinoma of the urinary bladder metastasizing to a solitary fibrous tumor of the pleura are described. No similar cases have been found in the accessible literature. These cases are discussed and the findings are correlated with the data of the literature. PMID- 12754637 TI - Transitional cell carcinoma arising in the gastric remnant following gastrocystoplasty: a case report and review of the literature. AB - Urinary bladder augmentation with segments of the stomach (gastrocystoplasty), small bowel, or large intestine (enterocystoplasty) improves capacity and compliance in patients with bladder dysfunction. Although malignant complications of enterocystoplasty have been reported, the risk of malignancy in the setting of gastrocystoplasty is not known. We describe the case of a 73-year-old woman who developed a transitional cell carcinoma associated with transitional cell metaplasia and dysplasia of the gastric epithelium 14 years following gastrocystoplasty. To our knowledge, this is the first reported case of a malignant complication of this surgical procedure. We conclude that patients who have undergone gastrocystoplasty are at an increased risk for the development of malignancy in the neobladder and require close long-term follow-up, similar to patients who have undergone enterocystoplasty. PMID- 12754638 TI - Leiomyosarcoma at the site of an ileal neobladder: a heretofore unreported occurrence. AB - We report a case of leiomyosarcoma arising from the intestinal side of an orthotopic ileal neobladder in a male patient 2 years after surgery for papillary urothelial carcinoma of the bladder. PMID- 12754639 TI - [Analysis of the emergency system of the Spanish neurosurgical services]. AB - OBJECTIVES: 1)To study the alert system (on call duty) in the Spanish neurosurgical departments and main issues of the emergency neurosurgical attention; 2) To establish common sense criteria for a thorough review of the system; 3) To analyse the effects of the current alert system on neurosurgical planning in Spain from the point of view of aging of Spanish neurosurgeons. MATERIAL AND METHODS: An interview containing 11 items related to the main aspects of the alert system was developed. The interview was sent to 51 departments. Answer from 42 of them were received. Neurosurgical departments were categorized in two different patterns to analyse the results: 1 st )according to the size of the staff. Three different types were observed:Type A (more than 10 neurosurgeons);Type B (between 6 and 10), and Type C (less than 6). 2 nd ) according to qualification of the personnel receiving the patient in the emergency area:Type I (the patient is received by paramedic personnel);Type II (trainee sends patient to specialist);Type III (training doctor orders diagnostic procedures) and Type IV (neurosurgeon orders diagnostic procedures and decides next actions). RESULTS: Type A departments :This group is formed by 8 departments. Alerts are always mix (at home or in hospital). There are usually 3 neurosurgeons on call (sometimes 4). Staff personnel is on call duty at the hospital 5 days a month plus 5-6 days at home. In 38% of the hospitals, department is type I-II (the patient is initially attended by paramedic personnel, not responsible for diagnostic or treatment). Surgical emergencies account for 0.54 a day as a rule and 12.75 patients require neurosurgeon's attention every day. In 75% of the hospital the neurosurgeon must also read neuroimaging procedures and suture scalps in 37%. Next day, neurosurgeon on call is off duty almost routinely. Type B :it was formed by 21 services. Alert are always mix (at home or in hospital). There are usually two neurosurgeons on call. Staff personnel are on call 4-5 days a month at the hospital and 5-6 days at home. The patient is initially attended by a doctor starting diagnosis in 85% of the cases (types III and IV).They operate 0.52 times a day,attending 6.19 patients/day as a mean. Neurosurgeon must read neuroimaging procedures in 57%. They do not repair scalps (except for a 5%) and personnel is off duty next day routinely. Type C :Nine (9) departments were grouped under this lining. Staff is always on call at home. There is usually only one neurosurgeon (sometimes two). Staff perform 10-12 alerts at home along the month. In 4 of the departments they must duplicate the number of days as far as there are two of them on-call. Patients are attended initially as type III-IV in 100% of the cases (doctor asks for diagnostic tools and decides therapy in 78%). Neurosurgeon on-call receives 4 5 phone-calls a day. Surgical emergencies account for 0.34 a day and 1.84 patients a day are attended directly by them. They never suture scalps. They seldom are off-duty next day. Aging of the Spanish neurosurgeons (389 interviewed). Most important issues are: 152 neurosurgeons will be 55 yo between 2003 and 2008, so they can ask for leaving on-call duties. 121 neurosurgeons will be 65 yo between 2009 and 2013. CONCLUSIONS: 1 st ) Alert system, mostly in bigger departments, is disproportioned and does not to fit to reality. 2 nd ) Emergency is worse organized in bigger hospitals. Besides, neurosurgeon on-call is not properly consulted. 3 rd ) Almost routinely neurosurgeons are offduty the day after alert in hospital. 4 th ) 152 neurosurgeons are necessary in the next five years to maintain the current system, which is obviously unviable. 5 th ) Most of the finishing trainees (residents) should tolerate "on-call contracts"in bigger departments to sustain the actual system. 6 th )For future planning, Neurosurgical National Committee must offer 20 training places a year to fill jubilees to happen from 2009 on. Finally we believe that the current alert system cannot be maintained any more, so Health Administratt system cannot be maintained any more, so Health Administrations must develop a profound reform. In this sense, the role that Local Neurosurgical Societies must play is essential. PMID- 12754640 TI - [Anatomical landmarks and surgical limits in the suboccipital transmeatal approach to the acoustic neuroma]. AB - INTRODUCTION: To completely remove the intracanalicular portion of the acoustic neuroma through the retrosigmoid approach, we must open the posterior wall of the internal auditory canal (IAC). Therefore, drilling the IAC is one of the key steps we need to take in the transmeatal surgical approach. Nevertheless, there are no clear anatomical landmarks to identify structures such as the semicircular canals, the jugular bulb or air cells. The individual anatomical variations and those caused by the tumour itself make preoperative evaluation essential if we wish to avoid complications such as deafness, cerebrospinal fluid leakage, bleeding and air embolism. OBJECTIVE: We describe here the personal experience of the senior author (EU) in drilling the posterior wall of the IAC, with special reference to the anatomical landmarks and surgical limits in the suboccipital approach to the intracanalicular portion of the acoustic neuromas. MATERIAL AND METHODS: This work is based on anatomical data obtained from drilling human temporal bones obtained from cadavers, along with our experience with 20 patients who were operated on for acoustic neuroma using Samii's technique. RESULTS: We did not operate on any purely intracanalicular neurinomas using this approach. Two tumors were grade II (up to 20mm in diameter), 12 were grade III and 6 were grade IV. We did not drill far enough in any of these cases to be able to see the fundus of the IAC, which was confirmed by postoperative CT. Despite this, the tumor was considered to be completely removed in 17 cases. There was no mortality and we has no major complications as a result of drilling the IAC such as cerebrospinal fluid leakage or air embolism. we cannot guarantee that hearing loss of postoperative deafness, which were the norm except in one case of grade II, were caused by nervous, ischemic or labyrinthine lesions. CONCLUSION: In our material it was not possible to completely expose the IAC fundus using a retrosigmoid approach without injury to labyrinth. The areas in which the risk of secondary complications is greatest when drilling are the inferior wall and the IAC fundus. The medial extension of the suboccipital craniotomy makes drilling the intrameatal tumor exposure easier. There are no intraoperative landmarks to locate the petrous structures while drilling the IAC except for those provided by the surgeon's own experience. PMID- 12754641 TI - [Analysis of cerebrospinal fluid related complications (hydrocephalus, fistula, pseudomeningocele and infection) following surgery for posterior fossa tumors]. AB - INTRODUCTION: Hydrocephalus, cerebrospinal fluid (CSF) leak, pseudomeningocele and CSF infection are potential complications related to surgical treatment of posterior fossa tumors. The objectives of this study were to review the incidence of such complications and to identify contributing factors related to them. MATERIAL AND METHODS: This study is based on a retrospective review of the medical records of 71 consecutive patients who underwent posterior fossa surgery for a tumor between the period January 1997 and December 2001. Postoperative hydrocephalus was defined as enlargement of the ventricles and the subsequent clinical worsening requiring surgical treatment. Criteria for CSF leakage were:observed leak of CSF through the wound, rhinorrhea or otorrhea. Pseudomeningocele was determined when there was a large epidural CSF collection diagnosed in the postoperative period or by magnetic resonance imaging performed at least three months after posterior fossa surgery. Finally, CSF infection was defined on clinical ground and positive biochemical examination, but not necessarily positive cultures. RESULTS: The series included 84 operations for resection of posterior fossa tumors on 71 patients. There were CSF related complications in 31% (26/84)with the following detailed incidence: 9.5% (8/84) postoperative hydrocephalus; 14.3% (12/84) CSF leak, 7.1% (6/84) pseudomeningocele; 8.3% (7/84) CSF infection. The mortality rate is 5.9% (5/84). The tumor size was the only statistically significant factor associated with the occurrence of CSF related complications (mean 39.43 mm, SD 18.51 mm vs.29.80 mm, SD 14.12 mm, p=0.015). In the subgroup of patients, in which hydrocephalus was managed preoperatively, the election of an external ventricular drain vs.other strategies (subcutaneous reservoir, definite shunt or endoscopic third ventriculostomy) was associated with a higher occurrence of CSF related complications (p=0.006). The mortality rate was associated with age (mean 63.60 years, SD 5.86 years vs.49.18 years, SD 16.39 years; p=0.002). The occurrence of CSF related complications also influenced mortality (p=0.030), particularly postoperative hydrocephalus (p< 0.001). Inpatient hospital stay was longer in the subgroup of patients who developed CSF related complications (p=0.002). CONCLUSIONS: Tumor size was the only factor associated with the development of CSF related complications after surgery for posterior fossa tumors. In the subgroup of patients in which hydrocephalus was surgically treated preoperatively, the election of an external ventricular drain compared to other surgical modalities was associated with a higher rate of CSF related complications. The development of such complications, particularly hydrocephalus, was related with mortality. PMID- 12754642 TI - [Treatment of intracranial germ cell tumours and other tumours of the pineal region]. AB - The management of patients with central nervous system germ-cell tumours is evolving, and a definitive standard has not been achieved. A large amount of data indicate that radiotherapy alone results in long-term relapse free survival rates of about 90% in patients with germinoma. Various prospective trials evaluated the results of combinations of chemotherapy and reduced dose and/or volume radiotherapy. The survival rates of combined treatment approaches were similar to the rates achieved with craniospinal radiotherapy alone. Nevertheless, the relapse rates were probably higher due to the significant number of relapses that arouse outside the volume treated with radiotherapy. Additional studies are necessary to determine the appropriate radiotherapy volumes and the role of combined treatments. Chemotherapy alone results in high relapse rates and can not be recommended. Mature teratomas are benign germ cell tumours that can be controlled with complete surgical resection in over 90% of cases. Non-germinoma germ cell tumours are a heterogeneous group of tumours that includes very aggressive tumours such as mixed and pure choriocarcinomas, yolk sac tumours, and embryonal carcinomas; and tumours with intermediate aggressiveness such as mixed tumours with germinoma and teratoma, immature teratomas and teratomas with malignant transformation. Both radiotherapy alone and chemotherapy alone result in quite low rates of tumour control and current treatment approaches include chemotherapy and radiotherapy, with surgical removal of the tumour in some patients. Pineocytomas are benign tumours that are controlled in most cases by complete surgical resection or partial surgical resection and local field irradiation. Current treatment approaches for pineoblastomas include surgery, chemotherapy, and craniospinal irradiation with a local boost. Chemotherapy alone was used to delay irradiation in infants with very little success. PMID- 12754644 TI - [Orbital meningocystocele]. AB - Encephalocele is defined as a protrusion of cranial contents beyond the normal confines of the skull. Encephaloceles occur with an incidence of approximately 1 in 3,000-10,000 births and are categorized based on location in the skull. Orbital meningoencephalocele is a rare congenital abnormality caused by a defect of the cranio-orbital bones that usually manifests soon after birth as a soft mass associated with exophthalmos. We present a case of a giant orbital meningocele presenting as an orbital mass in a 15 months-old girl. Preoperative diagnosis was confirmed by CT-scan and magnetic resonance imaging. PMID- 12754643 TI - [Cranial Procedures without shaving. A 1-year experience at the Hospital Sao Joao. ]. AB - Scalp shaving has been a ritual usually said to make easier orientation and surgical access and also to prevent local infection. However, this last aspect has been challenged in several publications and progressive evidence on favourable psychological effects of non-shaved patients has accumulated. The authors review 328 intracranial surgeries performed without scalp shaving from 1 October 1999 to 30 September 2000 in the Department of Neurosurgery of Hospital de S.Joao. All patients were included except those undergoing approaches, such as transphenoidal external ventricular drainages, removal of collocated non-organic materials and surgery for infections. The technique was implemented in patient having CSF shunting only during the second half of the study period. Antibiotic prophylaxis was the same previously used in the Department. The infection rate was 1.82%. The authors comment the disadvantages and benefits of the technique, remarking positive psychological effects for the patient. PMID- 12754645 TI - [Multiseptated arachnoid cyst treated with fenestration after valvular insufficiency in an adult. ]. AB - Treatment of symptomatic arachnoid cysts is based on two procedures: cyst fenestration versus derivation of CSF. Multiseptated cysts represent a very special group. We present the case of a 75 year old woman with a symptomatic multiseptated arachnoid cyst, developing subacute bleeding in one of the cavities. Final diagnosis was obtained after MRI. CSF derivation became insufficient with clear improvement after fenestration and communication to subarachnoid space. PMID- 12754646 TI - [The 11th reunion of the International Society of Leksell Gamma-rays Radiosurgery]. PMID- 12754650 TI - Hypointensity in T2-weighted images of the basal ganglia in solvent-exposed patients with multiple sclerosis: clinical, MRI and CSF characteristics. AB - Several studies have indicated an association between MS and organic solvent exposure. Our objective was to analyse differences regarding cerebrospinal fluid (CSF) properties, magnetic resonance imaging (MRI) features and cerebral metabolites, measured by proton spectroscopy (1H-MRS), in 20 patients with spontaneous multiple sclerosis (MS) and in 20 patients with MS after solvent exposure; 15 healthy subjects served as controls. CSF examinations were retrospectively reviewed from the medical files. There were no significant differences in the CSF regarding pleocytosis, spinal-serum albumin ratio or mean extended IgG index. However, T2-weighted images of the solvent-exposed MS patients showed more hypointenseareas in the basal ganglia. Hypointensity on T2 weighted images of the basal ganglia in the solvent-exposed MS patients may correspond to neurodegeneration and could be an early event in MS. PMID- 12754651 TI - How many camels are there in Italy? Cognitive estimates standardised on the Italian population. AB - OBJECTIVE: We provided the standardisation of a new Cognitive Estimation Task (CET). PARTICIPANTS: the test was administered to 175 healthy subjects. RESULTS: performance on the Cognitive Estimation Task (CET) is associated with gender (where women show poorer performance than men) and education (where more highly educated individuals show better performance compared to individuals with lower levels of education). However,CET performance is not associated with age. DISCUSSION: the lack of age effects on the CET may be explained by the task dependence on "crystallised intelligence", which is less affected by healthy adult ageing than "fluid intelligence". PMID- 12754652 TI - Neurophysiological features in relation to clinical signs in clinically diagnosed corticobasal degeneration. AB - We investigated the association between clinical and neurophysiological characteristics in patients with a clinical diagnosis of probable corticobasal degeneration (CBD), and searched for neurophysiological features supporting the diagnosis in life. Ten patients with clinically probable CBD underwent comprehensive neurological evaluation and brain MRI. Long latency reflexes (LLR), upper limb somatosensory (SEP) and motor evoked (MEP) potentials were recorded. The mini-mental state examination (MMSE), the phonemic verbal fluency test (PVFT) and the De Renzi ideomotor apraxia test were also performed. Polygraphic EEG was performed in the six patients with myoclonus. The SEP N30 frontal component was absent bilaterally in four patients, was absent on the left side in one, and had increased latency in other three. MEPs were abnormal in four patients (three had prolonged central motor conduction time, one of whom also had increased MEP threshold, and one had increased MEP threshold). All six patients with myoclonus had enhanced LLRs at rest, which were also of abnormally increased amplitude during motor activation; latencies were generally shorter than in classic cortical reflex myoclonus. On back-averaging, no EEG spikes time-locked to EMG activity were found in any myoclonus patient. Five patients were demented by MMSE, eight had ideomotor apraxia scores in the ideomotor apraxia range and five had defective verbal fluency. Brain MRI revealed asymmetric cortical atrophy in all patients, particularly evident frontoparietally. Neurophysiological techniques, particularly LLR, can assist CBD diagnosis especially in patients with myoclonus. Patients with evident parkinsonism had greater SEP N30 (frontal) abnormalities, while most patients with marked paresis had slower MEP times. PMID- 12754653 TI - Short-term longitudinal evaluation of cerebral blood flow in mild Alzheimer's disease. AB - The aim of this study was to estabilish whether subtle changes in regional cerebral blood flow (rCBF) can be detected in patients with mild Alzheimer's disease (AD) over a period as short as three months. Aprospective evaluation of rCBF changes with HMPAO SPECT at baseline and after 3 and 6 months was carried out. Standard clinical dementia tests were performed in parallel. The study enrolled 13 patients with mild probable AD from an outpatient memory clinic. SPECT data collected at baseline and after three months were compared using Statistical Parametric Mapping (SPM). Significant clusters of rCBF decrement in the medial part of the temporal lobe in both hemispheres, in the orbital part of the right frontal lobe and in the inferior part of the right parietal lobe were observed. No significant differences were found using a region of interest (ROI) analysis. After six months SPM analysis showed larger clusters of significant decrement in the same areas. ROI analysis was now sensitive to the rCBF changes and significant differences were shown in all brain regions except the temporal lobes. SPM analysis of SPECT data can detect significant changes in rCBF in mild AD over a short time interval. This method might be of potential advantage for the early diagnosis of AD and for its differentiation from stable cognitive impairments. PMID- 12754654 TI - Occupational head injury and subsequent glioma. AB - We report the case of a policeman who suffered a severe head injury to the right temporoparietal lobe while driving a police car. Four years later, the patient developed a neoplasm at the precise site of the meningocerebral scar. Histological examination confirmed a glioblastoma multiforme adjacent to the dural scar. Radiological documentation of the absence of tumor at the time of injury, exact localization of the neoplasm in the injured cerebral area, and latency of the cancer supported the hypothesis of a causal relationship with brain trauma. Physicians faced with brain neoplasms in adults should carefully investigate the patient's personal history of head trauma. When a relationship with occupational head injury is probable, reporting of suspect occupational illness is compelling. PMID- 12754655 TI - Myasthenia gravis with thymoma and autoimmune haemolytic anaemia. A case report. AB - Myasthenia gravis is an autoimmune disorder that affects the neuromuscular junction and leads to weakness of the skeletal muscles. Associated autoimmune diseases such as systemic lupus erythematosus, rheumatoid arthritis and pernicious anaemia are present in approximately 5% of the myasthenic patients. This report presents a 64-year-old man with autoimmune haemolytic anemia associated with myasthenia gravis and thymoma. The patient developed a severe Coomb's positive autoimmune haemolytic anaemia, which was resistant to treatment with large doses of prednisone. Haemolytic anaemia entered remission one month following thymectomy, and the patient has maintained a normal haemoglobin and a negative Coomb's test without the need for steroid or immunosuppressive therapy. In conclusion, thymectomy may induce a striking improvement of therapyresistant autoimmune haemolytic anemia in patients with MG and thymoma, but in terms of remission, a long follow-up is needed as autoimmune diseases can show spontaneous fluctuations. PMID- 12754656 TI - Multiple spinal meningiomas after tamoxifen therapy: a case report. AB - We describe a rare case of multiple spinal meningiomas and evaluate the possible relationship with tamoxifen treatment. We observed a 74-year-old woman who showed a spastic paraparesis gradually developed in the last year. The patient underwent left mastectomy for a breast cancer ten years earlier and was treated with tamoxifen for four years after surgical intervention. Magnetic resonance imaging revealed three spinal meningiomas at C6-C7, D6-D7 and D9 levels. Taking in account the tumor-inducing properties of tamoxifen and the extreme rarity of multiple spinal meningiomas, we suggest that tamoxifen may be the cause play a role in the genesis of the spinal meningiomatosis in the observed patient. Therefore, we propose the long-term clinical and neurological surveillance of patients who assumed tamoxifen, even for a short time, in order to survey the possible appearance of secondary tumours. PMID- 12754657 TI - Syringomyelia following Listeria meningoencephalitis: report of a case. AB - A case of symptomatic syringomyelia which appeared six years after Listeria meningoencephalitis is described. Chronic spinal arachnoiditis, as shown by standard MRI and dynamic phase contrast (PC) cine-MRI, may occur after spinal infection and is likely the cause of syringomyelia. To our knowledge, there are no previous reports of delayed spinal complications following Listeria monocytogenes infection. The possibility of developing syringomyelia should be always considered in any patient with a history of central nervous system infection. PMID- 12754659 TI - Characterization of microbial activities and U reduction in a shallow aquifer contaminated by uranium mill tailings. AB - A characterization of the Shiprock, NM, uranium mill tailing site focused on the geochemical and microbiological factors governing in-situ uranium-redox reactions. Groundwater and aqueous extracts of sediment samples contained a wide concentration range of sulfate, nitrate, and U(VI) with median values of 21.2 mM, 16.1 micro M, and 2.7 micro M, respectively. Iron(III) was not detected in groundwater, but a median value of 0.3 mM in sediment extracts was measured. Bacterial diversity down gradient from the disposal pile reflected the predominant geochemistry with relatively high numbers of sulfate- and nitrate reducing microorganisms, and smaller numbers of acetogenic, methanogenic, nitrate dependent Fe(II)-oxidizing, Fe(III)-reducing, and sulfide-oxidizing bacteria. In aquifer slurry incubations, nitrate reduction was always preferred and had a negative impact on sulfate-, Fe(III)-, and U-reduction rates. We also found that sulfate-reduction rates decreased sharply in the presence of clay, while Fe(III) reduction increased with no clear impact on U reduction. In the absence of clay, iron and sulfate reduction correlated with concentrations of Fe(III) and sulfate, respectively. Rates of U(VI) loss did not correlate with the concentration of any electron acceptor. With the exception of Fe(III), electron donor amendment was largely unsuccessful in stimulating electron acceptor loss over a 2-week incubation period, suggesting that endogenous forms of organic matter were sufficient to support microbial activity. Our findings suggest that efforts to accelerate biological U reduction should initially focus on stimulating nitrate removal. PMID- 12754660 TI - Distribution of exopolymeric substances in the littoral sediments of an oligotrophic lake. AB - Bacteria and algae release exopolymeric substances (EPS) that perform a wide range of important functions in aquatic and terrestrial systems. In this study we measured EPS in sediments at nine littoral sites around a shallow oligotrophic basin, and tested whether the concentration and composition of EPS was related to sediment characteristics. The concentrations of both loosely bound (colloidal) and tightly bound (capsular) EPS carbohydrates ranged up to ~800 micro g glucose equiv. cm(-2) and were well within the range of concentrations reported from marine intertidal flats, where EPS play an important role in stabilizing sediments, affecting nutrient exchanges between sediments and the water column, feeding benthic invertebrates, and sequestering and increasing the transfer of contaminants to food webs. Proteins were an important component of the EPS in these littoral sediments, with protein:carbohydrate ratios of approximately 0.4. In summer, the concentrations of most EPS fractions were positively related ( P < 0.05) to the porewater and organic matter content of the sediments. Capsular EPS concentrations were lower in the fall, with a simultaneous increase in colloidal proteins but not in colloidal carbohydrates. This suggests that the carbohydrates in this colloidal EPS may be more labile than the proteins. Our results suggest that exopolymeric substances could be an important, but neglected, component of littoral sediments in lakes. PMID- 12754661 TI - Detection of eubacterial 3-hydroxy-3-methylglutaryl coenzyme a reductases from natural populations of actinomycetes. AB - Three natural populations of actinomycetes were investigated by PCR for the presence of type I 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG CoA), a gene associated with isoprenoid biosynthesis. The populations were obtained from an agricultural site (69 isolates), a coastal salt marsh (220 isolates), and a desert soil (96 isolates). A set (34) of standard actinomycete reference strains were also investigated. The target gene was only detected in 5 of the 419 actinomycetes screened, which represented 4 from the coastal salt marsh and one reference strain. The isolates that contained the gene were taxonomically diverse (4 Streptomyces spp. and 1 Nocardia sp.). These results suggest that type I HMG CoA containing pathways are rare in actinomycetes and their distribution within actinomycetes populations is not random. PMID- 12754675 TI - Neuropsychological impairments and changes in emotional and social behaviour following severe traumatic brain injury. AB - Changes in emotional and social behaviour are relatively common following severe traumatic brain injury (TBI). Despite the serious consequences of these changes, little is known about the underlying neuropsychological deficits. In this study, we investigated which deficits might underlie these behavioural changes. The emotional and social behaviour of 17 patients with severe TBI was assessed with questionnaires, completed by the patient and a relative. Neuropsychological tests assessed recognition of emotional expressions, understanding of other people's mental states and cognitive fluency. Ratings from patients and relatives revealed changes in emotional and social behaviour after injury. Compared to matched healthy controls, the patients were impaired at recognising facial and vocal expressions of emotions, detecting social faux pas and nonverbal fluency. None of these impairments was significantly associated with the relatives' ratings of behavioural problems following TBI, although the correlation with detecting social faux pas was relatively high (r=-.61). PMID- 12754676 TI - The ESDQ: a new method of assessing emotional and social dysfunction in patients following brain surgery. AB - The Emotional and Social Dysfunction Questionnaire (ESDQ) has been designed to overcome some of the difficulties of inappropriately applying psychiatric based questionnaires to brain-damaged populations. Two-hundred and twenty-five patients were assessed following brain surgery (BS) using a self-rating patient version of the ESDQ and 211 of these patients were rated by their partner. A factor analysis using a varimax rotation and principal components analysis found the partner results to show eight factors including, Anger, Helplessness, Emotional Dyscontrol, Indifference, Inappropriateness, Fatigue, Maladaptive behaviour, and Insight. The analysis of the Self-rating questions revealed a similar profile, Anger, Emotional Dyscontrol, Helplessness, Inertia, Fatigue, Indifference, Inappropriate, and Euphoria. The scales based on the factors were subjected to discriminant analysis in which the BS patients were compared with a combination control group of neurosurgical outpatients and terminally-ill cancer patients, all of which were without cerebral complaints. The brain surgery results for the partner-rated and the Self-rated version of the ESDQ were compared with control ratings of 42 partners and 54 self-ratings, respectively. The analysis showed a significant discrimination for the Partner-rated version on each of the eight scales individually with an overall significant overall separation (Wilks Lambda=.903, chi=21.1, df=8, p=<.005). The Self-rated version showed less separation on an individual scale basis the Emotional Dyscontrol scale failing to show a significant separation. The overall difference on the Self-rating version was significant [Wilks Lambda=.908, chi=26.2, df=8, p=<.001). The levels of internal consistency of the questionnaire scales were found to be satisfactory (Alpha,.78 -.94). Also the relationship between ESDQ scales and standardised measures of aggression, anxiety, depression, and vigour (STAXI, HADS, and POMS) gave an indication of acceptable levels of concurrent validity. PMID- 12754677 TI - Verbal and nonverbal fluency performance before and after seizure surgery. AB - The utility of verbal (FAS) and nonverbal (Ruff Figural Fluency Test: RFFT) fluency tests for detecting deficits associated with focal seizures and surgical interventions was examined. The patients were 174 adults with intractable epilepsy who underwent epilepsy surgery: 152 temporal lobectomies and 22 frontal lobectomies. The results of the study suggest that the RFFT is somewhat superior to FAS in its ability to discriminate between frontal and temporal seizure foci, and is a useful component of preoperative neuropsychological batteries. Conversely, FAS appears more useful in detecting changes in neurocognitive outcome related to side of surgery. Controlling for postsurgical seizure outcome did not change the results, although continued seizures did have a deleterious effect on both FAS and RFFT, regardless of site of surgery. PMID- 12754678 TI - Fatigue in HIV/AIDS is associated with depression and subjective neurocognitive complaints but not neuropsychological functioning. AB - Fatigue and depressive symptoms are common in HIV-infection. The relationship between these symptoms and neuropsychological functioning is poorly understood, particularly in symptomatic infection/AIDS. This study examined the associations among fatigue, depressive symptoms, subjective neurocognitive complaints, and objective neuropsychological performance in HIV/AIDS. Sixty-eight men with HIV infection (27 adults with HIV-infection but not AIDS and 41 with AIDS diagnosis) completed a neuropsychological test battery and self-report measures of fatigue (Fatigue Severity Scale), depressive symptoms (Beck Depression Inventory), and subjective neurocognitive complaints (Patient's Assessment of Own Functioning). High levels of fatigue were endorsed by participants. Fatigue severity was related to depressive symptoms but not to AIDS diagnosis or medication status. Verbal learning and motor function was worse in participants with AIDS, but neuropsychological functioning was not significantly correlated with fatigue or depressive symptoms. Subjective neurocognitive complaints were predicted by both depressive symptoms and fatigue. Our results suggest that adults with fatigue and HIV-infection (with or without AIDS) should be screened for depression. Neither fatigue nor depressive symptoms appear to affect neuropsychological functioning in HIV/AIDS. Future research is needed to develop and evaluate instruments and methods to differentiate depression-related fatigue from fatigue that may reflect underlying medical disease. Such research will further the development of effective treatments for fatigue associated with HIV-infection. PMID- 12754679 TI - Alzheimer's disease versus normal ageing: a review of the efficiency of clinical and experimental memory measures. AB - This paper reviews research findings concerning memory performance in Alzheimer's disease (AD) and normal ageing. Studies using clinical (i.e., episodic) memory tests are compared with studies using various experimental memory paradigms (semantic memory, implicit memory, working memory), in order to determine their efficiency to differentiate between AD and normal ageing. In addition, attention is focused on early and preclinical AD. It is argued that traditional clinical memory tests alone are not best able at detecting AD at an early stage. More specifically, tasks calling upon semantic knowledge may aid to an earlier and more efficient assessment of AD. PMID- 12754681 TI - Normative data from the CANTAB. I: development of executive function over the lifespan. AB - The study of executive function within a developmental framework has proven integral to the advancement of knowledge concerning the acquisition and decline of higher skill processes. Still in its early stages, there exists a discontinuity in the literature between the exploration of executive capacity in young children and the elderly. Research of age-related differences utilising a lifespan approach has been restricted by the lack of assessment tools for the measurement of executive skills that are applicable across all age levels. This paper addresses these issues using the computer-based Cambridge Neuropsychological Test Automated Battery (CANTAB) to identify periods of development in executive capacities using a normative sample of 194 participants ranging in age from 8 to 64 years. Findings of executive function in children as young as 8 years of age were extended, with functional gains found in the efficiency of working memory capacity, planning and problem-solving abilities, between the ages of 15 and 19 years and again at 20-29 years of age. Cognitive flexibility was assessed at adult-levels in even the youngest children. Declines in performance on all tasks were revealed for the 50-64 year old sample, providing support for the vulnerability of executive skills to normal aging. PMID- 12754680 TI - 3MS normative data for elderly African Americans. AB - The Modified Mini-Mental State Examination (3MS) is an expanded and modified version of the Mini-Mental Status Exam (MMSE). Although the MMSE has achieved widespread clinical use as a brief cognitive screen, the utility of the measure to ascertain cognitive impairment is constrained by a limited set of abilities sampled, a narrow range of possible scores, floor and ceiling effects, and by a paucity of normative data for use with older adults from ethnically diverse backgrounds. Research demonstrates that the reliability, validity, and sensitivity of the 3MS are superior to that of the MMSE in detecting cognitive impairment. To date there has been minimal research investigating the usefulness of the 3MS with African American older adults. The current study examined the influence of demographic characteristics on the 3MS in a community-dwelling sample of 238 African American older adults (60-84 years). Age, gender, and education accounted for moderate amounts of variance in 3MS performance. Based on these results, normative tables for 3MS scores, stratified by age and with score adjustments for education and gender, are provided. PMID- 12754682 TI - Comparison of the psychosocial typology of children with below average IQ to that of children with learning disabilities. AB - This study investigates the subtypal patterns of psychosocial functioning of children with below average IQ (BAIQ) using the application of both Q-factor analysis and profile-matching. The results suggest that the psychosocial dimensions of children with BAIQ are quite similar to those of children with LD in a general sense. Many of the same subtypes were derived, and the proportions of children displaying normal, mild, and severe levels of psychopathology were not significantly different from those of children with LD. There were some minor differences, however: For example, children with BAIQ exhibited a greater tendency to display psychopathology with internalizing features. Consistent with previous research involving children with LD, there were no changes in either type or severity of psychopathology with advancing years. PMID- 12754683 TI - Comparison between three memory tests: cued recall, priming and saving closed head injured patients and controls. AB - Twenty closed-head injured (CHI) patients and 20 matched controls were tested with three different memory tasks: cued recall, word stem completion (WSC), and saving. Saving is defined as the advantage of relearning of a list of word pairs, in terms of the number of learning trials to the criterion of one errorless trial, over the original learning of the same list. It was predicted that CHI patients' explicit memory (i.e., cued recall), but not implicit memory (i.e., WSC), would be impaired. The question addressed in this study is whether the memory of CHI patients will be impaired when memory is tested with a saving task, with 2 weeks delay between original learning and relearning. The findings confirm impairment of CHI patients in explicit memory, although the learning rate is preserved. Implicit memory is preserved in CHI patients only when based on reactivation of preexisting knowledge, but not when dependent on forming new associations. Finally, the CHI patients, even after 2 weeks delay, demonstrated a significant saving in relearning old, as compared to new, pairs of words. The clinical contribution of this study is the delineation of those aspects of memory that are impaired and those that are preserved in CHI patients. The theoretical implications of the finding that memory could be preserved in CHI patients when measured by saving, are discussed in terms of the relationship between implicit memory and saving. PMID- 12754684 TI - Convergence of different versions of the continuous performance test: clinical and scientific implications. AB - The Continuous Performance Test has been used for the last 40 years to measure sustained attention or vigilance in many different populations. Different versions of the test have been developed, but little is known about how similar these tests are, and to what extent performance on different versions of these tests overlaps. In order to examine convergence of the different versions of the CPT, three different CPTs were administered in both the Auditory and Visual Sensory Modalities. Subjects were selected from consecutive admissions to adolescent acute care units at a private psychiatric hospital (n=100). Auditory test modalities uniformly elicited poorer performance than visual tests, while each set of task demands consistently elicited differences in performance. Despite the high test-retest reliability of the individual subtests, the average correlation between tests was r=.42, with the average correlation between visual tests at r=.48 and the average correlation between the auditory tests was r=.45. The correlations within task demands across sensory modalities ranged from a low of.37 to a high of.52. Controlling for IQ did not influence the correlations to a substantial degree. These data suggest different versions of the CPT are correlated with each other at a level consistent with construct validity, but that they do not constitute alternate forms of the same test. PMID- 12754686 TI - Environmental sample correlation with clinical and historical data in a friction product exposure. AB - Asbestos has been widely used in the past as a component in friction products. A unique setting of dust exposure to such products occurs when individuals are involved with refabrication of the worn components. It is of interest whether asbestos fibers are released from friction materials in the use phase and the postlife phase or if the mineral fibers are converted to a nonasbestos form. In the present study, an individual whose primary work activity had centered on clutch refabrication was evaluated for possible dust-related diseases. Tissue analysis revealed the presence of large numbers of asbestos fibers as well as ferruginous bodies. These particulates were characterized as to composition as well as dimensions. Clutches of the type typically used by this individual were obtained and material was rinsed from their surface for comparative purposes. Materials that were freed from the surface and reviewed by analytical transmission electron microscopy were clearly identifiable as chrysotile asbestos. The composition of the clutch material was compared with the components found in the patient's lung tissue. The conclusion from this comparison was that individuals exposed to friction materials under similar circumstances should use appropriate caution. Appreciable exposure to asbestos can occur and this exposure can be sufficiently high to result in disease. PMID- 12754687 TI - Nasal lavage method in the monitoring of upper airway inflammation: seasonal and individual variation. AB - A noninvasive and reliable method is needed to investigate causal relationship between exposure to bioaerosols in occupational and indoor environments and adverse health effects. As an essential part of the method development, the individual variation as well as seasonal and gender differences in the concentrations of inflammatory mediators in nasal lavage (NAL) fluid were studied. NAL was performed in 10 healthy volunteers every other week for a year. Concentrations of nitric oxide, assessed as nitrite, interleukin (IL)-1 beta, IL 4, IL-5, IL-6, and tumor necrosis factor alpha (TNF alpha) in NAL fluid were measured. The NAL sampling was minimally invasive and well tolerated and no side effects were observed among the studied subjects. Low concentrations of nitrite, TNFalpha, IL-1 beta, IL-4, and IL-6 were detected in the NAL samples of the studied subjects. Within-subject variation in the concentrations of inflammatory mediators in the NAL fluid was at its lowest during the wintertime. Moreover, differences between individuals and genders were statistically significant. In summary, the individual variation in the basal levels of measured inflammatory markers is low, whereas differences between individuals are considerable. Thus, in the studies evaluating upper airway effects of occupational or environmental exposure, the method is most suitable in settings where comparison can be made using test subjects as their own controls. PMID- 12754688 TI - Chronological changes in electrolyte levels in arterial blood and bronchoalveolar lavage fluid in mice after exposure to an edemagenic gas. AB - Detection of acute lung injury is important if therapeutic medical countermeasures are to be used to reduce toxicity in a timely manner. Indicators of injury may aid in the eventual treatment course and enhance the odds of a positive outcome following a toxic exposure. This study was designed to investigate the effects of a toxic exposure to the industrial irritant gas phosgene on the electrolyte levels in arterial blood and bronchoalveolar lavage fluid (BALF). Phosgene is a well-known chemical intermediate capable of producing life-threatening pulmonary edema within hours after exposure. Four groups of 40 Crl:CD-1(ICR)BR male mice were exposed whole-body to either air or phosgene at a concentration x time (c x t) amount of 32-42 mg/m(3) (8-11 ppm) phosgene for 20 min (640-840 mg x min/m(3)). BALF from air- or phosgene-exposed mice was taken at 1, 4, 8, 12, 24, 48, or 72 h postexposure. After euthanasia, the trachea was excised, and 800 microl saline was instilled into the lungs. The lungs were washed 5x. Eighty microliters of BALF was placed in a cartridge and inserted into a clinical i-STAT analyzer. Na(+), Cl(-), K(+), and ionized Ca(2+) were analyzed. Arterial blood electrolyte levels were also analyzed in four additional groups of air- or phosgene-exposed mice. The left lung was removed to determine wet weight (WW), an indicator of pulmonary edema. Na(+) was significantly higher in air than in phosgene-exposed mice at 4, 8, and 12 h postexposure. Temporal changes in BALF Cl(-) in phosgene mice were not statistically different from those in the air mice. Both Ca(2+) and K(+) were significantly higher than in the air-exposed mice over 72 h, p < or = 0.03 and p < or = 0.001 (two-way analysis of variance, ANOVA), respectively. Significant changes in BALF K(+) and Ca(2+) occurred as early as 4 h postexposure in phosgene, p < or = 0.005, versus air-exposed mice. Over time, there were no significant changes in arterial blood levels of Na(+), Cl(-), or Ca(2+) for animals exposed to air versus phosgene. However, arterial K(+) concentrations were significantly higher, p < or = 0.05, than in air-exposed mice across all time points, with the highest K(+) levels of 7 mmol/L occurring at 8 h and 24 h after exposure. Phosgene caused a time-dependent significant increase in WW from 4 to 12 h, p < or = 0.025, compared with air-exposed mice. These data demonstrate that measuring blood K(+) levels 1 h after exposure along with BALF Na(+), K(+), and Ca(2+) may serve as an alternate indicators of lung injury since both K(+) and Ca(2+) follow temporal increases in air-blood barrier permeability as measured by wet weight. PMID- 12754689 TI - The fate of antioxidant enzymes in bronchoalveolar lavage fluid over 7 days in mice with acute lung injury. AB - Characterization of lung injury is important if timely therapeutic intervention is to be used properly and successfully. In this study, lung injury was defined as the progressive formation of pulmonary edema. Our model gas was phosgene, a pulmonary edemagenic compound. Phosgene, widely used in industry, can produce life-threatening pulmonary edema within hours of exposure. Four groups of 40 CD-1 male mice were exposed whole-body to either air or a concentration x time (c x t) amount of 32-42 mg/m(3) (8-11 ppm) phosgene for 20 min (640-840 mg x min/m(3)). Groups of air- or phosgene-exposed mice were euthanized 1, 4, 8, 12, 24, 48, or 72 h or 7 days postexposure. The trachea was excised, and 800 micro l saline was instilled into the lungs and washed back and forth 5 times to collect bronchoalveolar lavage fluid (BALF). The antioxidant enzymes glutathione peroxidase (GPx), glutathione reductase (GR), superoxide dismutase (SOD), total glutathione (GSH), and protein were determined at each time point. Phosgene exposure significantly enhanced both GPx and GR in phosgene-exposed mice compared with air-exposed mice from 4 to 72 h, p < or = 0.01 and p < or = 0.005, respectively. BALF GSH was also significantly increased, p < or = 0.01, from 4 to 24 h after exposure, in comparison with air-exposed. BALF protein, an indicator of air/blood barrier integrity, was significantly higher than in air-exposed mice 4 h to 7 days after exposure. In contrast, BALF SOD was reduced by phosgene exposure from 4 to 24 h, p < or = 0.01, versus air-exposed mice. Except for protein, all parameters returned to control levels by 7 days postexposure. These data indicate that the lung has the capacity to repair itself within 24-48 h after exposure by reestablishing a functional GSH redox system despite increased protein leakage. SOD reduction during increased leakage may indicate that barrier integrity is affected by superoxide anion production. PMID- 12754690 TI - The temporal profile of cytokines in the bronchoalveolar lavage fluid in mice exposed to the industrial gas phosgene. AB - Diagnosis of an exposure to airborne toxicants can be problematic. Phosgene is used widely in industry for the production of many synthetic products, such as polyfoam rubber, plastics, and dyes. Although nearly 100% of the gas is consumed during processing, there is the potential problem of accidental or even intentional exposure to this irritant/choking agent. Exposure to phosgene has been known to cause latent life-threatening pulmonary edema. A major problem is that there is a clinical latency phase from 3 to 24 h in people before irreversible acute lung injury occurs. Assessment of markers of acute lung injury after a suspected exposure would be useful in developing rational treatment strategies. These experiments were designed to assess bronchoalveolar lavage fluid (BALF) for the presence of the early markers of exposure to phosgene in mice from 1 to 72 h after exposure. Separate groups of 40 CD-1 male mice (Crl:CD 1(ICR)BR) weighing 29 +/- 1 g were exposed whole-body to either air or a concentration x time (c x t) amount of 32 mg/m(3) (8 ppm) phosgene for 20 min (640 mg x min/m(3)). BALF from air- or phosgene-exposed mice was taken at 1, 4, 8, 12, 24, 48, and 72 h postexposure. After euthanasia, the trachea was excised, and 800 micro l saline was instilled into the lungs and washed 5x. BALF was assessed for interleukin (IL)-4, IL-6, tumor necrosis factor (TNF) alpha, IL 1alpha, macrophage inflammatory protein (MIP)-2, and IL-10. At 4 h postexposure, IL-6 was 15-fold higher for phosgene-exposed mice than for the time-matched air exposed control group. At 8 and 12 h, IL-6, IL-1beta, MIP-2, and IL-10 were significantly higher in phosgene-exposed mice than in time-matched air-exposed controls, p < or = 0.05 to p < or = 0.001, whereas TNF alpha reached peak significance from 24 to 72 h. IL-4 was significantly lower in the phosgene exposed mice than in the air-exposed mice from 4 to 8 h after exposure. These data show that BALF is an important tool in assessing pro- and anti-inflammatory markers of phosgene-induced acute lung injury and that knowledge of these temporal changes may allow for timely treatment strategies to be applied. PMID- 12754691 TI - Effects of di-n-butylamine on the respiratory system of Wistar (WU) rats after subchronic inhalation. AB - Aim of the study was to investigate the potential toxic effects of di-n butylamine (DBA), a known skin and eye irritating compound, on the respiratory tract after inhalation exposure for up to 91 days in male and female rats [Crl:(WI)WU BR]. To check whether and to what degree the no-observed-(adverse) effect level (NO(A)EL) decreases with increasing study duration, serial sacrifices were performed after 3 and 28 days, respectively. Based on two dose range-finding studies, the concentrations for this study were determined with 0 (clean air), 50, 150, and 450 mg/m(3). Animals were exposed for 3 days (6 h/day) 28, and 91 days (5 days/wk, 6 h/day), respectively, and immediately sacrificed thereafter. The results show clear irritating effects only in the upper part of the respiratory tract, that is, the nasal cavities. While after 3 and 28 days effects were found only in the high-dose group, slight adaptive effects, expressed as mucous (goblet) cell hyperplasia, could be diagnosed in the medium- and low-dose groups after 91 days of exposure. Pathological changes were most prominent after 3 days of exposure. In the lung, only marginal effects could be observed (increased relative lung weight only in females of the high concentration after 28 days, slight, not statistically significant histopathological effects in the high concentration after 3 days, no effects on parameters of bronchoalveolar lavage fluid), while no effects were found in the remaining groups. PMID- 12754692 TI - Quantification of continuous glass filaments on eclipse cigarettes retrieved from the test market. AB - ECLIPSE cigarettes utilize a special form of continuous glass filament (CGF) as an insulator around the carbon heat source. The average numbers of CGFs on the external barrel and cigarette filter end were determined subsequent to manufacture, subsequent to real-world consumer handling and subsequent to simulated consumer handling. The following were not statistically significantly different: the average number of CGFs on the external barrel of cigarettes retrieved from the test market compared to the average external barrel counts from cigarettes subsequent to manufacture or when subjected to simulated consumer handling, and the average number of CGFs on the external barrel of cigarettes subsequent to manufacture compared to the average external barrel counts from cigarettes subjected to simulated consumer handling. The average number of CGFs on the filter end of cigarettes retrieved from the test market was statistically significantly higher than average cigarette filter end counts from cigarettes subsequent to manufacture. The average number of CGFs on the cigarette filter end of cigarettes retrieved from the test market was statistically significantly lower than average cigarette filter end counts from cigarettes subjected to simulated consumer handling. Overall, results from this study suggest that consumer handling does increase the average numbers of CGFs on the external surfaces of the cigarette. Further, the results of this study demonstrate that for the purpose of CGF quantification, the simulated consumer handling protocol used in this study (i.e., based on laboratory measurements of forces) is a reasonably good model for actual consumer handling of cigarettes. Based on the minimal number of CGFs that could be transferred to the smoker and the deposition pattern governed by their physical characteristics, the potential to deposit CGFs from these cigarettes to the lungs of smokers is extremely remote. Therefore, no convincing information exists to suggest that smokers would be exposed to CGFs from any ECLIPSE-related source at a biologically significant level. PMID- 12754693 TI - Introduction to the special issue on international clinical psychology. AB - We briefly describe the content of the six research articles selected by peer review for this, the first special issue of the Journal of Clinical Psychology devoted to international clinical psychology. Two of the articles address general scientific issues-illusory mental health and a theory of anorexia nervosa-not considered specific to any particular cultural setting. One article examines social anxiety in three different Western societies. One considers the development of clinical psychology in a specific country, Spain. The final two articles consider two clinical problems-sexual dysfunction and Type-I diabetes within two different contexts in India, one Hindu, the other Moslem. The introduction concludes with some general comments on the history and present status of clinical psychology as an international field. PMID- 12754694 TI - Cloning the clinician: a method for assessing illusory mental health. AB - Self-report mental health scales cannot distinguish between genuine mental health and the facade or illusion of mental health created by psychological defenses. Shedler, Mayman, and Manis (1994) demonstrated that experienced clinicians can differentiate genuine from illusory mental health using the Early Memory Test, and that illusory mental health may be a risk factor for medical illness. This article describes the development of the Early Memory Index (EMI), a formal scoring system for the Early Memory Test, and presents evidence for its reliability and validity. The EMI makes the technique for assessing illusory mental health accessible to a wide range of investigators, including investigators who lack extensive clinical training. PMID- 12754696 TI - Social anxiety in three Western societies. AB - The present study investigates whether empirical data support the notion that people in Western societies do not differ with regard to social anxiety. Social anxiety in Dutch students (N = 425) was compared with that experienced by students in the United States (N = 440) and Turkey (N = 349). Social anxiety was operationalized with the Inventory of Interpersonal Situations (IIS; Van Dam Baggen & Kraaimaat, 1987, 1999, 2000), which measures two aspects of social anxiety, i.e., discomfort in social situations and frequency of social responses. The original Dutch version of the IIS (IOA) was translated into a U.S. English version (the IIS) and a Turkish version (KADE). First, it was shown that the American, Dutch, and Turkish versions of the IIS measured the same construct in terms of factor structure. Second, American students generally appeared more socially anxious than did the Dutch and Turkish students, whereas the latter's social anxiety slightly surpassed that of Dutch subjects. The American students also showed fewer social skills than both other groups, who did not differ in this respect. The results are somewhat more differentiated with respect to the domains of social anxiety, and they are discussed in terms of cross-cultural differences and their implications for clinical practice. PMID- 12754695 TI - The cognitive-orientation theory of anorexia nervosa. AB - The major goal was to explore the cognitive-motivational dynamics of anorexia in terms of the cognitive-orientation (CO) theory (Kreitler & Kreitler, 1982). CO is a comprehensive theory of behavior that assumes that behavior is a function of a cognitively shaped motivational disposition and performance. The study deals with the motivational disposition for anorexia. It focused on examining whether beliefs of four types (about self, goals, norms, and reality) concerning themes relevant for anorexia (defined in pretests) identify correctly anorectics. All participants were women 15 to 18 years old: 58 anorectics (35 restricting, 23 binge eating/purging) and 59 matched healthy controls. All were administered a background-information questionnaire and the CO-Anorexia questionnaire assessing beliefs about 30 themes. The results showed that the themes formed 5 clusters defined by foci, such as dissociation from reality, the body, drives or emotionality, and identified significantly the anorectics of each type and the healthy controls. A brief CO questionnaire was developed. Discussion centered on the similarity of the identified themes to some of those discussed by others, on the pathogeneity of the CO of anorexia, and on outlining a blueprint of a theory of anorexia. PMID- 12754697 TI - Clinical psychology in Spain. AB - A general view of the field of clinical psychology in present-day Spain is offered here. The field has experienced an enormous development over the past two decades. Many journals and specialized societies have been established, and a large number of professionals now are working in the field, most of them in private practice, but some in medical centers belonging to the Spanish national health service. This great expansion seems due mainly to the creation of a degree in psychology(1968) and the continuous flow of students demanding training in clinical subjects. A theoretical orientation toward a cognitive-behavioral approach seems to dominate, closely followed by those choosing a dynamic orientation. A large group of professionals dealing with health problems are reinforcing their idiosyncratic profile among the clinical psychology. A new study program to become a specialist in our field recently has been implemented and is just entering its initial stages (Internship in Psychology-PIR); it represents a significant improvement in the curriculum of the clinical psychology. All of these facts seem to prove the great vitality of this field in present-day Spain. PMID- 12754698 TI - Sexual dysfunction in single males: a perspective from India. AB - Various personal, familial, and socio-cultural factors play a crucial role in the development of sexuality and sexual practices. They often influence the occurrence and maintenance of sexual dysfunctions. Therapies with individuals with sexual dysfunctions often emphasize the involvement of both the partners. Issues related to single males, especially in the Indian context, have not been explored. The present study is a retrospective analysis of clinical case records of single males who reported sexual dysfunctions between the years 1990 to 2000. Thirty-eight clinical case records were analyzed for the demographic details, nature of the problems, and interventions provided with the aim of exploring the reported symptomatology, precipitating and maintaining factors, prevalent beliefs about the causation of sexual dysfunction, and the outcome of interventions. The role of cultural and psychosocial issues is discussed and the need for research in this area is emphasized. PMID- 12754699 TI - Role of family in the management of Type-I diabetes: an Indian experience. AB - The role of the family in the management of Type-I diabetes is gaining recognition. In countries where the adolescent is dependent on the family for medical needs, the family's role is all the more important. At times, when the family is uncooperative, the care of the adolescent is hampered, making psychosocial intervention even more difficult. The following case study illustrates the difficulty encountered while working with a young diabetic belonging to an Indian family. The issues related to parental role and management of diabetes-related problems are discussed. PMID- 12754700 TI - Variations of the human glucocorticoid receptor gene (NR3C1): pathological and in vitro mutations and polymorphisms. AB - Glucocorticoid (GC) resistance can occur in a number of diseases. It can be either generalized (i.e., familial glucocorticoid resistance) or localized (i.e., asthma). In many cases, a reason for this resistance to steroids lies with mutations or polymorphisms present in the glucocorticoid receptor gene (GR/NR3C1) that belongs to a large family of nuclear receptors. A number of GC-resistant cell lines have been isolated in vitro, some of which arose or may have arisen in vivo. These and the mutations defined in them are included in this review as well as mutations engineered in plasmids and expressed in vitro. It also lists polymorphisms and the individual studies where association-related studies have been performed. NR3C1 is located on chromosome 5q31 and contains 10 exons that code for a 777 amino acid protein. There are two naturally occurring isoforms of the NR3C1, GRalpha (functional) and GRbeta (no hormone-binding ability). A total of 15 missense, three nonsense, three frameshift, one splice site, and two alternative spliced mutations have been reported in the NR3C1 gene associated with glucocorticoid resistance as well as 16 polymorphisms. Mutation and polymorphism data for NR3C1 will soon be found on the newly created locus specific database. PMID- 12754701 TI - Mutations in the molybdenum cofactor biosynthetic genes MOCS1, MOCS2, and GEPH. AB - Molybdenum cofactor deficiency in humans results in the loss of the activity of molybdoenzymes sulfite oxidase, xanthine dehydrogenase, and aldehyde oxidase. The resultant severe phenotype, which includes progressive neurological damage leading in most cases to early childhood death, results primarily from the deficiency of sulfite oxidase. All forms of molybdenum cofactor deficiency are inherited as autosomal recessive traits. The cofactor is an unstable reduced pterin with a unique four-carbon side chain, synthesized by a complex pathway that requires the products of at least four different genes (MOCS1, MOCS2, MOCS3, and GEPH). Disease-causing mutations have been identified in three of these genes: MOCS1, MOCS2, and GEPH. MOCS1 and MOCS2 have a bicistronic architecture; i.e., each gene encodes two proteins in different open reading frames. The protein products, MOCS1A and B and MOCS2A and B, are expressed either from different mRNAs generated by alternative splicing or by independent translation of a bicistronic mRNA. The gephyrin protein, encoded by a third locus, is required during cofactor assembly for insertion of molybdenum. A total of 32 different disease-causing mutations, including several common to more than one family, have been identified in molybdenum cofactor-deficient patients and their relatives. PMID- 12754702 TI - Human Gene Mutation Database (HGMD): 2003 update. AB - The Human Gene Mutation Database (HGMD) constitutes a comprehensive core collection of data on germ-line mutations in nuclear genes underlying or associated with human inherited disease (www.hgmd.org). Data catalogued includes: single base-pair substitutions in coding, regulatory and splicing-relevant regions; micro-deletions and micro-insertions; indels; triplet repeat expansions as well as gross deletions; insertions; duplications; and complex rearrangements. Each mutation is entered into HGMD only once in order to avoid confusion between recurrent and identical-by-descent lesions. By March 2003, the database contained in excess of 39,415 different lesions detected in 1,516 different nuclear genes, with new entries currently accumulating at a rate exceeding 5,000 per annum. Since its inception, HGMD has been expanded to include cDNA reference sequences for more than 87% of listed genes, splice junction sequences, disease-associated and functional polymorphisms, as well as links to data present in publicly available online locus-specific mutation databases. Although HGMD has recently entered into a licensing agreement with Celera Genomics (Rockville, MD), mutation data will continue to be made freely available via the Internet. PMID- 12754703 TI - Mutant NDUFV2 subunit of mitochondrial complex I causes early onset hypertrophic cardiomyopathy and encephalopathy. AB - Respiratory chain complex I deficiencies represent a genetically heterogeneous group of diseases resulting from mutations in either mitochondrial or nuclear DNA. Combination of denaturing high performance liquid chromatography and sequence analysis allowed us to show that a 4-bp deletion in intron 2 (IVS2+5_+8delGTAA) of the NDUFV2 gene (encoding NADH dehydrogenase ubiquinone flavoprotein 2) causes complex I deficiency and early onset hypertrophic cardiomyopathy with trunk hypotonia in three affected sibs of a consanguineous family. The homozygous mutation altering the consensus splice-donor site of exon 2 resulted in 70% decreased NDUFV2 protein and complex I deficiency. While mutation in a number of genes encoding complex I subunits essentially result in neurological symptoms, this first mutation in NDUFV2 is strikingly associated with cardiomyopathy, as previously observed in the unique case of NDFUS2 mutations. PMID- 12754704 TI - Single base substitutions at the initiator codon in the mitochondrial acetoacetyl CoA thiolase (ACAT1/T2) gene result in production of varying amounts of wild-type T2 polypeptide. AB - Initiator codon mutations are relatively uncommon and less well characterized compared to other types of mutations. We identified a novel initiator codon mutation (c.2T>C) heterozygously in a Japanese patient (Patient GK30) with mitochondrial acetoacetyl-CoA thiolase (T2) gene deficiency (ACAT1 deficiency); c.149delC was on the other allele. We examined translation efficiencies of nine mutant T2 cDNAs harboring one-base substitutions at the initiator methionine codon using in vivo transient expression analysis. We found that all the mutants produced wild-type T2 polypeptide, to various degrees (wild type (100%) > c.1A>C (66%) > c.2T>C, c.3G>C, c.3G>T (22%) > c3G>A, c.1A>G (11%) > c.2T>A, c.2T>G, c.1A>T (7.4%)). T2 mRNA expression levels in Patient GK08 (a homozygote of c.2T>A) and Patient GK30 fibroblasts, respectively, were almost the same as in control fibroblasts, when examined using semiquantitative PCR. This means that initiator codon mutations did not affect T2 mRNA levels. We propose that all one base substitutions at the initiator methionine codon in the T2 gene could be mutations, which retain some residual T2 activity. PMID- 12754705 TI - Mutation analysis in patients with N-acetylglutamate synthase deficiency. AB - N-acetylglutamate synthase (NAGS) is the key enzyme for the regulation of the hepatic urea cycle and is also highly expressed in kidney and gut. The reaction product, N-acetylglutamate, is an allosteric activator of carbamylphosphate synthetase 1 in the liver, catalyzing the initial step of ammonia detoxification. NAGS deficiency is a rare inborn error of metabolism inherited as an autosomal recessive trait leading to hyperammonemia. Using homology search based on genetic information of ascomycetes, we identified the human gene for NAGS on chromosome 17q21.31. There is a distinct pattern of organospecific expression of transcripts in liver, small intestine, and kidney similar to the other mitochondrially located enzymes of the urea cycle. The encoded 534 amino acid polypeptide has a consensus sequence for a 49 amino acid mitochondrial leader peptide. We identified private mutations of the NAGS gene in patients with severe early onset of clinical symptoms (IVS3-2A>T, c.1306_1307insT, c.971G>A/W324X, c.1289T>C/L430P, c.1299G>C/E433S, c.1450T>C/W484R), as well as in a case with late onset (c.835G>A/A279P). Four out of seven mutations were detected on exon 6. This is the first report of mutation analysis in a series of families affected with deficiency of NAGS. Molecular analysis of patients and reliable antenatal diagnostics for affected families are now feasible. PMID- 12754707 TI - Pseudoexon activation in the DMD gene as a novel mechanism for Becker muscular dystrophy. AB - We report the characterization of two deep intronic mutations in the Duchenne muscular dystrophy (DMD) gene of two unrelated Becker muscular dystrophy (BMD) patients, causing the aberrant inclusion of a pseudoexon in the mature transcripts. These two mutations were identified by the use of RT-PCR on transcripts isolated from muscle. The first abnormally large transcript resulting from a 58-bp insertion between exon 62 and exon 63 was identified in a BMD patient with mental retardation. The origin of this transcript was a mutation in intron 62 (IVS62-285A>G), which resulted in the occurrence of a high quality donor splice site. The IVS25+2036A>G in intron 25 was identified in a subclinical BMD patient with high CK levels. The mutation reinforces the strength of a pre existing acceptor splice site, resulting in activation of an intronic pseudoexon of 95 bp. By using DHPLC, the patient's mother was found to be a somatic mosaic. The insertion of these newly recognized extra exons leads to premature termination codons, but we could observe that some degree of normal splicing was taking place in both patients. The detection of these residual full length transcripts is consistent with the clinical presentation and dystrophin analyses. This is the first report of pseudoexon activation as a mechanism for Becker muscular dystrophy, and this reveals further the diversity of genetic abnormalities causing BMD. PMID- 12754706 TI - Molecular and phenotypic heterogeneity in mitochondrial trifunctional protein deficiency due to beta-subunit mutations. AB - The mitochondrial trifunctional protein (TFP) is a multienzyme complex of the fatty acid beta-oxidation cycle. It is composed of four alpha-subunits (HADHA) harboring long-chain enoyl-CoA hydratase and long-chain L-3-hydroxyacyl-CoA dehydrogenase (LCHAD) and four beta-subunits (HADHB) harboring long-chain 3 ketoacyl-CoA thiolase (LKAT). Mutations in either subunit can result in TFP deficiency with reduced activity of all three TFP enzymes. We characterize 15 patients from 13 families with beta-subunit mutations by clinical, biochemical, and molecular features. Three clinical phenotypes are apparent: a severe neonatal presentation with cardiomyopathy, Reye-like symptoms, and early death (n=4); a hepatic form with recurrent hypoketotic hypoglycemia (n=2); and a milder later onset neuromyopathic phenotype with episodic myoglobinuria (n=9). Maternal HELLP syndrome occurred in two mothers independently of the fetal phenotype. Mutational analysis revealed 16 different mutations, the majority being missense mutations (n=12). The predominance of missense mutations and the milder myopathic phenotype are consistent. Based upon homology to yeast thiolase that has been characterized structurally, the mutation localization within the protein correlates with the clinical phenotype. Outer loop mutations that are expected to alter protein stability less were only present in milder forms. The degree of reduction in thiolase antigen also correlated with the severity of clinical presentation. Although TFP deficiency is highly heterogeneous, there is genotype-phenotype correlation. PMID- 12754708 TI - De novo SCN1A mutations are a major cause of severe myoclonic epilepsy of infancy. AB - Severe myoclonic epilepsy of infancy (SMEI or Dravet syndrome) is a rare disorder occurring in young children often without a family history of a similar disorder. The earliest disease manifestations are usually fever-associated seizures. Later in life, patients display different types of afebrile seizures including myoclonic seizures. Arrest of psychomotor development occurs in the second year of life and most patients become ataxic. Patients are resistant to antiepileptic drug therapy. Recently, we described de novo mutations of the neuronal sodium channel alpha-subunit gene SCN1A in seven isolated SMEI patients. To investigate the contribution of SCN1A mutations to the etiology of SMEI, we examined nine additional SMEI patients. We observed eight coding and one noncoding mutation. In contrast to our previous study, most mutations are missense mutations clustering in the S4-S6 region of SCN1A. These findings demonstrate that de novo mutations in SCN1A are a major cause of isolated SMEI. PMID- 12754709 TI - Molecular detection of novel WFS1 mutations in patients with Wolfram syndrome by a DHPLC-based assay. AB - Wolfram syndrome (WS) is a recessively inherited mendelian form of diabetes and neurodegeneration also known by the acronym DIDMOAD from the major clinical features, including diabetes insipidus, diabetes mellitus, optic atrophy, and deafness. Affected individuals may also show renal tract abnormalities as well as multiple neurological and psychiatric symptoms. The causative gene for WS (WFS1) encoding wolframin maps to chromosome 4p16.1 and consists of eight exons, spanning 33.44 Kb of genomic DNA. In this study we report on the mutational analysis of the WFS1 coding region in 19 Italian WS patients and 25 relatives, using a DHPLC-based protocol. A total of 19 different mutations in WFS1 were found in 18 of 19 patients (95%). All these mutations, except one, are novel, preferentially located in WFS1 exon 8, and include deletions, insertions, duplications, and nonsense and missense changes. In particular, a 16 base-pair deletion in WFS1 codon 454 was detected in five different unrelated nuclear families, being the most prevalent alteration in this Italian group. Nine neutral changes and polymorphisms were also identified. Overall, this study represents the molecular characterization of the largest cohort of Italian WS patients and carriers studied so far, and increases the number of identified WFS1 allelic variants worldwide. PMID- 12754710 TI - A novel splice site mutation in the TRIM37 gene causes mulibrey nanism in a Turkish family with phenotypic heterogeneity. AB - Mulibrey nanism (muscle-liver-brain-eye nanism; MUL) is an autosomal recessively transmitted disease characterized by severe growth delays of prenatal onset caused by mutations in the TRIM37 gene. Recent studies on the subcellular localization revealed that the TRIM37 (KIAA0898) protein is located in peroxisomes. Therefore, MUL has been classified as a new peroxisomal disorder. Up to now, four mutations have been reported, all of which lead to frameshifts and truncated proteins. In this study, mutation screening was performed for the coding region of the TRIM37 gene in a Turkish family by means of RT-PCR and direct cDNA sequencing. We have identified a novel mutation resulting in a frameshift cosegregating within the family. Finally, we report on the presence of novel splice variants observed in lymphoblastoid cells and muscle tissue of normal subjects and patients. PMID- 12754711 TI - An ABCA4 genomic deletion in patients with Stargardt disease. AB - Stargardt disease (STGD1) segregates with mutations in the ABCA4 (ABCR) locus. However, mutations of the ABCA4 coding region detected by sequencing account for only 66-80% of disease chromosomes. We hypothesized a potential contribution of otherwise undetected genomic rearrangements of the ABCA4 region. To investigate this hypothesis, we performed genomic Southern analysis on samples from 96 STGD families in which we had identified either one or no ABCA4 mutations by conventional methods. Among 192 chromosomes evaluated, we found one deletion (0.52%), IVS17-905_IVS18+35del, that spans 1,030 bp and eliminates exon 18 of ABCA4. By conceptual translation, this alteration creates an in-frame deletion of 30 amino acids, G885_H915del, and cosegregates with the disease in this family, implying a disease-associated allele. STGD subjects with this deletion were found to have a second mutant ABCA4 allele, 2588G>C. DNA sequence analysis of the deletion junction revealed consensus DNA topoisomerase I sites at both breakpoints that may predispose to nonhomologous recombination. Using deletion specific PCR, we found the same allele in 2 of 308 STGD subjects (0.32%), in 1 of 96 age-related macular degeneration (AMD) subjects (0.52%), and in 2 of 480 (0.2%) individuals with no known eye diseases, but it was absent in a control group consisting of 96 individuals over age 60 and with normal eye examinations. In vitro biochemical studies of the cloned G885_H915del mutation revealed diminished expression, suggesting that partial deletion of the putative nucleotide-binding domain I leads to either misfolding or defective membrane interactions and eventually reduces the protein function in the retinopathy affected subjects. Our experiments suggest that genomic alterations contribute to only a small fraction of retinopathy-associated alleles. PMID- 12754712 TI - A multiplex methylation PCR assay for identification of uniparental disomy of chromosome 7. AB - Uniparental disomy of chromosome 7 (UPD7) is associated with abnormal phenotypic effects because of inappropriate expression of imprinted genes on chromosome 7. Based on the differential methylation of the promoter region of the imprinted PEG1/MEST locus at 7q32, we designed a multiplex methylation PCR (mPCR) assay to rapidly distinguish UPD7 from biparental inheritance of chromosome 7. Primers were designed to produce different sized PCR amplicons based on the parent of origin-specific methylation at this locus; electrophoresis of PCR amplicons showed a 189-bp product from the methylated maternal allele and a 109-bp product from the unmethylated paternal allele. This mPCR assay correctly predicted the chromosome 7 imprinting status in normal control and UPD7 samples. Previous assays for UPD7 required genotyping of the proband and parents, or separate maternal- and paternal-specific mPCR reactions. The advantage of this assay is that parental samples are not required and that amplification of both alleles in the same reaction is simpler and provides an internal control. This multiplex mPCR assay will be useful in screening for UPD7 in patients with Silver-Russell syndrome (SRS; also Russell-Sliver syndrome, RSS), primordial growth retardation, and in patients with supernumerary marker chromosomes or chromosome rearrangements of chromosome 7 origin. PMID- 12754713 TI - Cultured fibroblasts as a tool for improvement of molecular analysis in ornithine transcarbamylase (OTC) deficiency. PMID- 12754715 TI - Interim analysis of continuous long-term endpoints in clinical trials with longitudinal outcomes. AB - This paper discusses interim analysis for clinical trials where the primary endpoint is observed at a specific long-term follow-up time, but where repeated measures of the same outcome are also taken at earlier times. Methods are considered for improving the efficiency with which the long-term treatment difference is estimated, making use of information from shorter-term follow-up times. This approach to interim analysis has previously been studied for binary endpoints assessed at two time points during follow-up. Here we adapt and extend this methodology to include continuous endpoints assessed at an arbitrary number of follow-up times, making use of methods for analysing multivariate normal data subject to monotone missingness and unstructured mean and covariance relationships. The magnitude of efficiency gains obtained by using short-term measurements is considered, as well as how these gains depend on the number and timing of the short-term measurements. Sequential analysis of treatment differences is discussed, including the extent to which efficiency gains translate into reductions in the expected duration of a sequentially monitored trial. The methods are illustrated on a data set involving a placebo-controlled comparison of longitudinal cholesterol measurements. PMID- 12754716 TI - Estimates, power and sample size calculations for two-sample ordinal outcomes under before-after study designs. AB - Sample size calculations are given for comparing two groups of subjects, typically referring to active and non-active intervention groups, on an ordinal outcome in experiments where the subjects are measured before and after intervention. These calculations apply to log-odds models with random intercepts, treatment, time and treatment-by-time interaction terms, the latter being the term of interest. The assumed forms of the odds ratios are flexible, allowing for proportional odds, adjacent categories, or other conditional models for ordinal responses. Simulations studies show that, for given sample sizes, the nominal and actual powers of the proposed test are similar. PMID- 12754717 TI - Robustness of sample size re-estimation procedure in clinical trials (arbitrary populations). AB - In clinical trials, one of the main questions that is being asked is how many additional observations, if any, are needed beyond those originally planned. In a two-treatment double-blind clinical experiment, one is interested in testing the null hypothesis of equality of the means against one-sided alternative when the common variance sigma2 is unknown. We wish to determine the required total sample size when the error probabilities alpha and beta are specified at a predetermined alternative. Shih provided a two-stage procedure which is an extension of Stein's one-sample procedure, assuming normal response. He estimates sigma2 by the method of maximum likelihood via the EM algorithm and carries out a simulation study in order to evaluate the effective level of significance and the power. The author proposed a closed-form estimator for sigma2 and showed analytically that the difference between the effective and nominal levels of significance is negligible and that the power exceeds 1-beta when the initial sample size is large. Here we consider responses from arbitrary distributions in which the mean and the variance are not functionally related and show that when the initial sample size is large, the conclusions drawn previously by the author still hold. The effective coverage probability of a fixed-width interval is also evaluated. Proofs of certain assertions are deferred to the Appendix. PMID- 12754718 TI - Interval estimates of the probability of toxicity at the maximum tolerated dose for small samples. AB - Following on from the work of O'Quigley et al., we investigate the performance of interval estimates of the probability of toxicity following completion of a phase I clinical trial. Our particular focus is on very small sample sizes, not uncommon in phase I studies. Specifically, we study the situations for which the sample sizes are 12 or 16. Simulations are used to show that the coverage of the confidence intervals, even for very modest sample sizes, are close to nominal in most cases. Averaged over the range of situations considered, coverage rates are accurate for both sample sizes of 12 and 16. As for the larger sample size of 20, studied previously, it is possible to obtain further, albeit modest, improvements via the use of the Cornish-Fisher inversion. PMID- 12754719 TI - Age-conditional probabilities of developing cancer. AB - We propose an estimator of the probability of developing a disease in a given age range, conditional on never having developed the disease prior to the beginning of the age range. Our estimator improves the one described by Wun, Merrill and Feuer ( Lifetime Data Analysis 1998; 4, 169-186) that is currently used by the U.S. National Cancer Institute for the SEER Cancer Statistics Review. Both estimators use cross-sectional disease rates and provide an interpretation of these rates in terms of the age-conditional probability of developing disease in a hypothetical cohort. The difficulty of this problem is that rates are not available per person-years alive and disease free, but only per person-years alive. Wun et al. used ad hoc methods to handle this problem which did not properly account for competing risks, did not provide a measure of variability, and only allowed age ranges using prespecified 5-year age intervals. Here we solve the problem under a unified competing risks framework, which allows the calculation of the age-conditional probabilities for any age range. We generalize gamma confidence intervals to apply to our new statistic. Although our new method provides estimates which are numerically similar to that of Wun et al., this paper provides a comprehensive theoretical basis for estimation and inference about the age-conditional probability of developing a disease. PMID- 12754721 TI - A modified large-sample approach to approximate interval estimation for a particular intraclass correlation coefficient. AB - We consider the problem of constructing a confidence interval for the intraclass correlation coefficient in an interrater reliability study when both raters and subjects are random effects in a two-way random effects model. A simulation study is conducted to investigate and compare the interval coverage per cents of three methods of approximation: (i) Satterthwaite's two moments, the standard approach; (ii) modified three moments, a newer approach; and (iii) modified large sample, the newest approach for this particular problem. One-sided lower and two-sided bounds are examined. For the two-moment approach, coverage of confidence intervals can be understated for not only one-sided bounds but also two-sided bounds. Coverage of confidence intervals for the modified large-sample approach is either correct or conservative and provides narrower two-sided widths than the modified three-moment approach, which can understate coverage on one-sided and two-sided bounds for two or three raters. The competing methods are illustrated with data from a reliability study of four raters evaluating the number of decayed, missing, and filled surfaces in the permanent teeth of ten subjects. Overall, the modified large-sample approach provides the most satisfactory performance of the three methods. PMID- 12754720 TI - Likelihood-based confidence intervals for a log-normal mean. AB - To construct a confidence interval for the mean of a log-normal distribution in small samples, we propose likelihood-based approaches - the signed log-likelihood ratio and modified signed log-likelihood ratio methods. Extensive Monte Carlo simulation results show the advantages of the modified signed log-likelihood ratio method over the signed log-likelihood ratio method and other methods. In particular, the modified signed log-likelihood ratio method produces a confidence interval with a nearly exact coverage probability and highly accurate and symmetric error probabilities even for extremely small sample sizes. We then apply the methods to two sets of real-life data. PMID- 12754722 TI - Microarrays, pattern recognition and exploratory data analysis. AB - We explore the application of principal component based approaches to pattern recognition in microarray analysis. A comparative assessment is presented, based on predictive evaluation, following a review of some key methodology. On the basis of these results, we select principal component based linear discrimination for a further in-depth analysis. We are particularly interested in studying the use of principal component decomposition for the evaluation of the condition of estimation of the pooled covariance matrix. The results are used to give some guidance as to how principal components may be used as a data exploratory tool in the analysis of microarray data. Opportunities for further development are outlined as well as implications for wider statistical modelling of microarray data. PMID- 12754723 TI - A maximum likelihood approach for estimating the QT correction factor using mixed effects model. AB - Assessment of QT interval prolongation is often used for assessing the cardiac safety of a new drug. However, the correction of the QT interval for varying heart rates has potential bias due to various different correction factors. This article proposes a maximum likelihood (ML) approach for calculating the appropriate individual correction factor using the data. The data come from a study with 24 subjects participating in a 10 day multiple dose (NEW RX) placebo controlled cross-over trial with repeat ECGs obtained at baseline and at day 10. ML techniques were used to fit a random-effects model to observed QT and HR values for estimating the pooled and individual correction factors. QT(c) values using four correction factors (Bazett, Friderecia, pooled and individual) were investigated. The relative performance of the various correction factors are given in terms of variability and graphical techniques. The pooled correction factor was estimated to be 0.292 and the individual correction factors ranged from 0.19 to 0.41. The assessment of the treatment effect on QT(c) yielded inconsistent results. Bazett's factor indicated prolongation (6.55+/-1.20), Friderecia's factor indicated no change, while the pooled (-2.92+/-0.94) and individual (-2.82+/-1.00) factors showed a significant decrease. Graphical examination of individual QT(c) data showed a significant advantage in the use of individual correction factors versus Bazett's factor both in terms of sensitivity as well as reduction in bias. Use of individual correction factors is advocated for the assessment of possible drug-induced QT(c) prolongation. PMID- 12754724 TI - Hierarchical linear models for the development of growth curves: an example with body mass index in overweight/obese adults. AB - When data are available on multiple individuals measured at multiple time points that may vary in number or inter-measurement interval, hierarchical linear models (HLM) may be an ideal option. The present paper offers an applied tutorial on the use of HLM for developing growth curves depicting natural changes over time. We illustrate these methods with an example of body mass index (BMI; kg/m(2)) among overweight and obese adults. We modelled among-person variation in BMI growth curves as a function of subjects' baseline characteristics. Specifically, growth curves were modelled with two-level observations, where the first level was each time point of measurement within each individual and the second level was each individual. Four longitudinal databases with measured weight and height met the inclusion criteria and were pooled for analysis: the Framingham Heart Study (FHS); the Multiple Risk Factor Intervention Trial (MRFIT); the National Health and Nutritional Examination Survey I (NHANES-I) and its follow-up study; and the Tecumseh Mortality Follow-up Study (TMFS). Results indicated that significant quadratic patterns of the BMI growth trajectory depend primarily upon a combination of age and baseline BMI. Specifically, BMI tends to increase with time for younger people with relatively moderate obesity (25 BMI <30) but decrease for older people regardless of degree of obesity. The gradients of these changes are inversely related to baseline BMI and do not substantially depend on gender. PMID- 12754725 TI - Priorities for sleep research during the next decade. PMID- 12754726 TI - Evolution of the caregiving experience in the initial 2 years following stroke. AB - Relationships between stroke survivor and family caregiver factors and the caregiver's health-related quality of life (HRQL) and overall quality of life (QoL) were examined in 97 dyads during the first and second years after stroke. Compared to age- and sex-matched population norms, caregivers scored significantly lower on the mental subscales of HRQL, and differences were greater for women than for men. Caregiver characteristics (older age, less burden, and fewer physical symptoms) were associated with better HRQL (mental summary scale) in the first year, with similar findings in the second year. Moderate stroke survivor physical impairment and caregiver characteristics (younger age and better HRQL) were associated with better QoL in the first year. During the second year poorer caregiver physical and mental health and caring for a stroke survivor with communication difficulties were associated with diminished QoL. PMID- 12754727 TI - Relationship of social role quality to psychological well-being in women with rheumatoid arthritis. AB - The purpose of this study was to examine the mediating and moderating effects of women's social role quality on the psychological well-being of women with rheumatoid arthritis (RA). One hundred and fifty-six women with a diagnosis of RA (M age = 59, SD = 11) completed self-report measures of arthritis history, physical health, psychological well-being, and role quality. Hierarchical multiple regression analyses indicated that role quality mediated the effects of physical health on depression and purpose in life, moderated the effects of health on depression, and moderated the effects of pain on purpose in life. Women in poor health with high role quality were significantly less depressed than women in poor health with poor role quality. Women with high levels of pain and high role quality had more purpose in life than women with high levels of pain and low role quality. Despite difficulties with their physical health, women who had high role quality had higher levels of psychological well-being. Findings from this study may aid in the development of meaningful interventions to help women with RA manage their daily lives to optimize well-being. PMID- 12754728 TI - Detecting acute confusion in older adults: Comparing clinical reasoning of nurses working in acute, long-term, and community health care environments. AB - In an article on a previous study involving hospitalized older adults (McCarthy, 2003), it was argued that the theory of situated clinical reasoning explains why nurses often fail to recognize acute confusion. Further, the theory illuminates how nurses' perspectives toward health in aging affect the ways they regard and ultimately deal with older people in this particular clinical situation. The purpose of the current study was to challenge and refine the theory by exploring the influence of different care environments on clinical reasoning related to acute confusion. Following a period of participant observation, a purposive sample of 30 nurses, 10 each from a teaching hospital, a long-term facility, and a home care agency, participated in semistructured interviews. Dimensional analysis provided the methodological framework for data collection and interpretation. The results reinforce prior findings that the ability of nurses to recognize acute confusion and to distinguish it from dementia can be attributed to their personal philosophies about aging. Care environment was identified as a factor that influenced clinical reasoning in limited ways under certain conditions and within certain contexts. PMID- 12754729 TI - Determinants of physical activity and adherence to a 24-week home-based walking program in African American and Caucasian women. AB - The purposes of this study were to: (a) identify which determinants of physical activity among African American and Caucasian women predict adherence to a 24 week home-based walking program; and (b) explore differences between African American and Caucasian women. Participants were 153 working women who were sedentary at leisure. The program included an exercise prescription, instructions, and support. Background determinants included demographics, previous exercise experience, and social role influences. Intrapersonal determinants (self-efficacy, self-determinism) were measured at baseline and 24 weeks. Adherence was measured with heart rate monitors and logs. Adherence was significantly higher in Caucasians, those with less previous exercise experience, and those with higher self-efficacy. Findings suggest that adherence to a walking program is influenced by multiple factors including background and intrapersonal determinants of physical activity. PMID- 12754730 TI - Effect of a psychiatric diagnosis on nursing care for nonpsychiatric problems. AB - This study was designed to test how patients' psychiatric diagnoses would affect nursing care for medical problems. Sixty nurses were randomly assigned to three groups in this posttest-only experiment. Control group nurses read a vignette describing a man admitted with a possible myocardial infarction (MI). Nonpsychotic group nurses also read that the person was on alprazolam. Psychotic group nurses read that the person was on haloperidol, benztropine, lorazepam, trazadone, fluoxetine, and lithium. Psychotic group nurses estimated a decreased probability that the patient was having an MI and were less likely to respond to additional possible MI symptoms. An increased awareness of the potential to stereotype medical patients with a history of psychosis might assist nurses when providing care. PMID- 12754731 TI - Cross-cultural validation of the stages of the tobacco acquisition questionnaire and the decisional balance scale. AB - The first phase in a study of smoking behaviors of 11- to 14-year-old Taiwanese children was designed to ensure the reliability and validity of newly translated instruments. The stages of the tobacco acquisition questionnaire (STAQ) and the decisional balance scale (DBS) were translated into Chinese, then back-translated into English. The DBS was adapted based on input from a focus group with young adolescents, reviewed by a panel of experts and laypersons, and pilot-tested. The next step consisted of administering the instruments to 401 children (ages 11-14 years) for psychometric testing. Factor analysis yielded three components for the STAQ accounting for 57.8% of the total variance, with alphas of the subscales ranging from.85 to.92. The DBS had two components accounting for 59.3% of the total variance, with alphas of.87 and.90 for the subscales. Accuracy of the translated instruments was supported by the psychometric test results. PMID- 12754732 TI - An alternative paradigm for clinical nursing research: an exemplar. AB - Effectiveness research is undertaken to evaluate the effects of interventions in achieving desired outcomes when tested in the real-world conditions of everyday practice. Although the randomized clinical trial (RCT) is considered the gold standard for effectiveness research, its feasibility, generalizability, and the clinical utility of its results are being questioned. This state of the science prompted the call for a paradigm shift, characterized by alternative methods for clinical research. The alternative methods attempt to account for clinical realities when conducting research, with the goal of minimizing discrepancies in the perspective and assumptions underlying practice and research. In this article a theory-driven approach to intervention evaluation is presented as a viable alternative paradigm for clinical research. The application of this approach demands changes in four aspects of research: participant selection criteria, assignment to treatment options, delivery of the intervention, and selection of outcome measures. The changes are discussed at the conceptual level and illustrated with examples from an ongoing multisite study aimed at determining the usefulness of this theory-driven approach to intervention evaluation. PMID- 12754733 TI - An alternative view on "an alternative paradigm". PMID- 12754734 TI - The attractive Achilles heel of germ cell tumours: an inherent sensitivity to apoptosis-inducing stimuli. AB - Testicular germ cell tumours (TGCTs) are extremely sensitive to cisplatin containing chemotherapy. The rapid time course of apoptosis induction after exposure to cisplatin suggests that TGCT cells are primed to undergo programmed cell death as an inherent property of the cell of origin. In fact, apoptosis induction of germ cells in the testis is an important physiological mechanism to control the quality and quantity of the gametes produced. Although p53 protein is highly expressed in the majority of TGCTs, almost no p53 mutations have been detected. Interestingly, p53 overexpression is associated with loss of p21 and gain of mdm2 expression, which might indicate a partial loss in functionality of the p53 regulatory pathway in TGCTs. Besides p21, TGCTs often show low expression of other proteins involved in the regulation of cell cycle progression, such as the retinoblastoma protein and members of the INK4 family. It can be postulated that the deregulated G(1)-S phase checkpoint results in premature entry into the S phase upon DNA damage. In addition to Bcl-2 family members that are involved in the regulation of germ cell apoptosis in the normal testis via the mitochondrial death pathway, the Fas death pathway is also known to regulate apoptosis of germ cells in the testis. Since chemotherapy has been shown to activate the Fas death pathway and TGCTs co-express both Fas and its ligand FasL, TGCT cells might undergo apoptosis upon cisplatin treatment via autocrine or paracrine activation of the Fas system by FasL. The hypothesis suggested here is that the lack of cell cycle arrest following a cisplatin-containing treatment, together with the activation of the Fas death pathway and the mitochondrial death pathway, explains the rapid and efficient apoptosis of TGCT cells. Defining the mechanisms involved in the cisplatin sensitivity of TGCTs will provide tools to increase cisplatin sensitivity in other human tumours with acquired or intrinsic resistance. PMID- 12754735 TI - Deregulation of the RB pathway in human testicular germ cell tumours. AB - Deregulation of the RB pathway is shared by most human malignancies. Components upstream of the retinoblastoma tumour suppressor (pRB), namely the INK4 family of cyclin-dependent kinase (CDK) inhibitors, the D-type cyclins, their partner kinases CDK4/CDK6, and pRB as their critical substrate, are differentially targeted in diverse types of cancer. An 'unorthodox' spectrum of defects within this cascade occurs in testicular germ cell tumours (TGCTs), including silencing of pRB transcription, overexpression of cyclin D2, and loss of p18INK4c. To improve understanding of the role of this pathway in spermatogenesis, and its subversion in TGCTs, we examined immunohistochemical expression patterns of CDK4, p16INK4a, p15INK4b, and pRB, and established an in situ assay for cyclin D mediated phosphorylation of serine795, a phosphorylation event critical for neutralization of pRB's growth-restraining ability. pRB was expressed throughout adult spermatogenesis and was detectable in teratomas, but was absent or grossly reduced in carcinoma in situ (CIS) and most seminomas and embryonal carcinomas. Unexpectedly, we also found that pRB was absent from fetal human gonocytes, the candidate target cell for all types of TGCTs. Thus, rather than a tumorigenesis promoting loss of pRB, the lack of pRB in TGCTs likely reflects its developmental control. Widespread expression of p15INK4b, found in normal testes, was preserved in TGCTs. In contrast, p16INK4a was lost or reduced in large subsets of TGCTs. CDK4 was expressed in normal spermatogonia, CIS, and invasive TGCTs, as was serine795-phosphorylated pRB. Our data on expression of pRB support the plausible origin of TGCTs from fetal gonocytes, and the serine795 phosphorylation demonstrates that the cyclin D-dependent kinases are active, and neutralize pRB in spermatogonia and in those TGCTs that express pRB. We hope that this study will inspire further immunohistochemical applications of phosphospecific antibodies in pathology, and examination of the RB pathway defects in relation to curability of TGCTs. PMID- 12754736 TI - Laminin alpha1 chain in human renal cell carcinomas and integrin-mediated adhesion of renal cell carcinoma cells to human laminin isoforms. AB - In human tissues, the laminin (Ln) alpha1 chain shows a restricted and developmentally regulated distribution in basement membranes (BMs) of a subset of epithelial tissues, including those of renal proximal convoluted tubules. The present study investigated the distribution of the Ln alpha1 chain in renal cell carcinomas (RCCs) and oncocytomas as well as in xenografted tumours induced in nude mice with four characterized RCC cell lines. These cell lines were also used in cell adhesion studies with purified laminins. By immunohistochemistry it was found that the Ln alpha1 chain is widely present in the BMs of RCCs, all of the specimens presenting immunoreactivity. High-grade RCCs tended to contain more BM confined and stromal immunoreactivity than low-grade tumours, none of the grade 3 (G3) carcinomas being negative and all of the metastatic specimens showing partial or overall BM immunoreactivity. Double immunolabelling experiments showed that in RCC BMs but not in vessel walls, the Ln alpha1 chain was co-distributed with Ln alpha5, beta1, and beta2 chains, implying the presence of Ln-1/Ln-3 and Ln-10/Ln-11. In papillary RCCs, the Ln alpha1 chain co-localized with Ln-5. The oncocytomas lacked immunoreactivity for the Ln alpha1 chain. Xenografted tumours induced in nude mice showed BM-like deposition of the Ln alpha1 chain. In cell adhesion studies, mouse and human Ln-1 were equally effective in promoting cell adhesion of all RCC cell lines. For each cell line, Ln-10 and Ln-10/11 were equally effective adhesive substrates, all cell lines adhering more avidly to these laminins than to mouse or human Ln-1. As judged by inhibition assays employing specific integrin antibodies, adhesion of normal human renal proximal tubular epithelial (RPTE) cells and RCC cells from a G1 tumour to human Ln-1 was mediated mainly by alpha(6)beta(1) integrin, while only the G1 RCC cells adhered to mouse Ln-1 by using alpha(6)beta(1) integrin. For adhesion to Ln-10, RPTE cells and RCC cells from a G1 tumour used an unidentified beta(1) integrin. Cells from G3 tumours mainly used an alpha(3)beta(1) integrin complex for adhesion to mouse Ln-1 and to human Ln-1 and Ln-10. For all cells, adhesion to the Ln-10/11 mixture was mediated by an unidentified integrin complex or by other adhesion molecules. These results show that laminin trimers containing the alpha1 chain are, in contrast to oncocytomas, abundant in the BMs of RCCs. This is in keeping with their suggested origin from renal proximal tubular epithelium known for its capacity to produce the Ln alpha1 chain. The results also show that RCC cells utilize complex, mainly integrin alpha(3)beta(1)- and integrin alpha(6)beta(1) mediated, mechanisms for adhesion to laminins. The adhesion to Ln-1 changes from integrin alpha(6)beta(1) to integrin alpha(3)beta(1) upon increasing malignancy and, especially for Ln-10 and Ln-10/11, other adhesion molecules of non-integrin type may contribute to the adhesion. PMID- 12754737 TI - Analysis of genetic alterations, classified according to their DNA ploidy pattern, in the progression of colorectal adenomas and early colorectal carcinomas. AB - DNA aneuploidy is a biological marker of the oncogenic potential of colorectal adenomas. The accumulation of genetic alterations of cancer-related genes is also essential for colorectal carcinogenesis. However, it is unclear whether there is any relationship between these genetic alterations and the DNA ploidy of colon tumour cells in the progression of colorectal adenomas and early colorectal carcinomas. Here we have studied the DNA ploidy state and genetic alterations occurring in colorectal tumours using the crypt isolation technique. Crypts isolated from a total of 106 colorectal tumors (adenoma, 93; early carcinoma, 13) were examined using a combination of flow cytometric analysis of DNA content, polymerase chain reaction-microsatellite assay, and single-strand conformation polymorphism assay for evidence of chromosomal allelic imbalance (AI; 17p; 5q; 18q) or p53 gene mutation. In addition, we examined microsatellite instability (MSI) with BAT 26 primer sets. DNA multiploidy was infrequently detected in colorectal adenomas (15.1%), in contrast to early carcinomas (46.2%). There was a significant difference in the incidence of AI of chromosome 18q between diploid adenomas and aneuploid populations of multiploid adenomas (18.1% vs 57.1%, p = 0.0043). Mutation of p53 was also found more frequently in aneuploid populations of early multiploid colorectal carcinomas than in early diploid colorectal carcinomas (66.7% vs 0%, p = 0.021). MSI was found in only 2 of 93 adenomas, with no MSI detected in early colorectal cancers. The two MSI-positive adenomas were diploid. We subdivided multiploid adenomas into two groups: those with a low or a high DNA index (DI). The incidence of genetic alterations of high-DI adenomas did not differ from those of low-DI adenomas. Allelic imbalance involving loci on chromosome 18q and mutations of p53 seems to be associated with the progression of diploidy to multiploidy in colorectal tumours. On the other hand, MSI may be associated with the development of some diploid tumours. In addition, the incidence of genetic alterations in the colorectal adenomas that we examined appears to be independent of the tumour's DNA index. PMID- 12754738 TI - The zinc finger protein OZF (ZNF146) is overexpressed in colorectal cancer. AB - Overexpression of the OZF gene has previously been demonstrated in the majority of pancreatic cancers. However, because the stages of tumour progression in this disease are poorly defined, no conclusion could be drawn concerning the relationship between OZF overexpression and the course of tumour progression. In contrast, initiation and progression steps are well defined in colorectal cancer. Most colon cancers are believed to arise from polypoid adenomas as a result of the gradual accumulation of genetic alterations, allowing the study of genetic events in the early stages of neoplasia. Accordingly, we wanted to assess the frequency of OZF overexpression in this tumour type and the relationship between overexpression and disease stage. Twenty-five colon carcinoma specimens from different sites and at various stages were analysed by immunoblotting using a highly specific mouse monoclonal antibody. Each sample was compared with adjacent normal colonic mucosa. Complementary immunohistochemical analysis was also carried out on a commercially available tissue array to identify cells expressing OZF. Of the 25 tumours analysed by immunoblotting, 20 expressed higher levels of OZF protein than their adjacent normal mucosa. Immunohistochemistry revealed OZF expression in tumour cells of 56/59 carcinomas and occasionally in infiltrating lymphocytes but at lower levels. Little or no staining was observed in cells taken from normal or inflammatory colon specimens. In both immunoblot and immunohistochemistry experiments, no correlation was found between OZF expression and clinical parameters such as TNM classification, stage, localization and age. Immunostaining was observed in low-grade adenomas, indicating that OZF expression occurs very early in the course of tumour progression. OZF expression, infrequent or absent in normal colonic mucosa, is present in more than 80% of colorectal cancers. Dysregulation of the OZF gene is an early event that may be implicated in the genesis of colonic carcinoma and may therefore provide a potential therapeutic target. PMID- 12754740 TI - Absence of lymphangiogenesis and intratumoural lymph vessels in human metastatic breast cancer. AB - Early metastasis to lymph nodes is a frequent complication in human breast cancer. However, the extent to which this depends on lymphangiogenesis or on invasion of existing lymph vessels remains controversial. Although proliferating intratumoural lymphatics that promote nodal metastasis have been demonstrated in experimental breast tumours overexpressing VEGF-C, it has yet to be determined whether the same phenomena occur in spontaneous human breast cancers. To address this important issue, the present study investigated the lymphatics in primary human breast carcinoma (75 cases of invasive ductal and lobular breast cancer) by quantitative immunohistochemical staining for the lymphatic endothelial hyaluronan receptor LYVE-1, the blood vascular marker CD34, and the nuclear proliferation marker pKi67. None of the breast carcinomas was found to contain dividing lymph vessels, even in areas of active haemangiogenesis. Furthermore, the majority of non-dividing lymph vessels were confined to the tumour periphery where their incidence was low and unrelated to tumour size, grade or nodal status; rather, their density was inversely correlated with tumour aggressiveness as assessed by macrophage density (p = 0.009), and blood microvessel density (p = 0.05, Spearman Rank), as well as with distance from the tumour edge. Finally, a proportion of the peritumoural lymphatics contained tumour emboli associated with hyaluronan, indicating a possible role for LYVE-1/hyaluronan interactions in lymphatic invasion or metastasis. These results suggest that naturally occurring breast carcinomas invade and destroy lymph vessels rather than promoting their proliferation; that breast tumour lymphangiogenesis may not always occur at physiological VEGF-C levels; and that nodal metastasis can proceed via pre existing lymphatics. PMID- 12754739 TI - The angiogenic switch for vascular endothelial growth factor (VEGF)-A, VEGF-B, VEGF-C, and VEGF-D in the adenoma-carcinoma sequence during colorectal cancer progression. AB - Angiogenesis is essential for tumour growth and metastasis. It is controlled by angiogenic factors, one of the most important being vascular endothelial growth factor (VEGF)-A. Although its role has been demonstrated in many tumour types including colorectal carcinoma (CRC), the importance of the newer family members in adenoma, invasive tumour growth, and progression to a metastatic phenotype has been poorly characterized in CRC. The aim of this study was to determine the role and timing of the VEGF angiogenic switch during CRC progression. We measured the gene expression of VEGF ligands (VEGF-A, VEGF-B, VEGF-C, and VEGF-D) and their receptors (VEGFR-1, VEGFR-2, and VEGFR-3), in normal colorectal tissues (n = 20), adenomas (n = 10), and in CRC (n = 71) representing different Duke's stages using ribonuclease protection assay, semi-quantitative relative reverse transcriptase polymerase chain reaction, together with the pattern of their expression by immunohistochemistry. VEGF-A mRNA was the most abundant in colorectal tissue, followed by VEGF-B, VEGF-C, and VEGF-D. VEGF-A and VEGF-B mRNAs were significantly more abundant in adenomas (p = 0.0003 and p = 0.04 respectively) compared with normal tissues, while VEGF-A and VEGF-C were significantly increased in carcinomas compared with normal tissues (p = 0.0006 and p = 0.0009 respectively). A significantly greater amount of VEGF-C mRNA was present in carcinomas compared with adenomas (p = 0.03), whereas there was a significant reduction of VEGF-B in carcinomas compared with adenomas (p = 0.0002). VEGF-D mRNA was significantly more abundant in normal tissues than in adenomas (p = 0.0001) and carcinomas (p < 0.0001). In normal tissues distant from the primary tumour, there was a significantly greater amount of VEGF-A and VEGF-D mRNA in patients with Duke's B and Duke's C respectively, compared with Duke's A stage tumours (p = 0.04 and p = 0.01 respectively). Immunohistochemistry showed low basal levels of all ligands in histologically normal tissues and their expression in the epithelium of tumours reflected the levels of mRNA expression identified. VEGF-A and VEGF-C mRNA levels correlated significantly with tumour grade (p = 0.01 and p = 0.01 respectively) and tumour size (p = 0.001 and p = 0.01 respectively), but not with patient age, sex, presence of infiltrative margin, lymphocytic response, vascular invasion, Duke's stage, or lymph node involvement (p > 0.05). VEGF-B mRNA correlated with an infiltrative margin (p = 0.04) but no other clinicopathological variable, and expression of VEGF-D demonstrated no association with any parameter examined. VEGFR-1 was significantly correlated with tumour grade (p = 0.02), Duke's stage (p < 0.001), and lymph node involvement (p = 0.004), VEGFR-2 with lymph node involvement (p = 0.02), and VEGFR-3 did not correlate with any of the clinicopathological variables tested. These results suggest that VEGF-A and VEGF-B play a role early in tumour development at the stage of adenoma formation and that VEGF-C plays a role in advanced disease when there is more likelihood of metastatic spread. The finding of increased levels of VEGF-A and VEGF-D expression in normal tissues collected from a site distant from the primary tumour indicates changes in the surrounding tumour environment that may enhance the subsequent spread of tumour cells. PMID- 12754741 TI - Prognostic significance of BRCA1 expression in sporadic breast carcinomas. AB - BRCA1 is a tumour suppresser gene frequently mutated in familial breast cancer and thought to influence the progression of sporadic breast cancer. Decreased BRCA1 mRNA and protein expression has been identified in breast cancer cell lines and sporadic breast tumours. Here the prognostic significance of reduced BRCA1 protein expression is investigated in primary operable breast cancer. Immunohistochemical analysis was used to determine the level of BRCA1 protein expression in 100 breast cancers. BRCA1 expression was compared with known prognostic factors and survival to investigate its prognostic significance. BRCA1 nuclear expression was reduced by varying amounts in breast carcinomas. A progressive loss of BRCA1 expression correlated well with higher histological grade (p = 0.002) and an excess of medullary/atypical medullary/grade 3 ductal carcinomas (p = 0.0001). When adjusted for grade, patients with loss of BRCA1 expression had a significantly longer disease-free survival time. Loss of BRCA1 expression associated with high-grade breast tumours suggests that BRCA1 may play an important role in the pathogenesis of sporadic breast cancer. PMID- 12754742 TI - Differential expression and function of A20 and TRAF1 in Hodgkin lymphoma and anaplastic large cell lymphoma and their induction by CD30 stimulation. AB - A20 and TRAF1 are two anti-apoptotic components of the intracellular signalling pathway of the tumour necrosis factor receptor (TNFR) family. Induction of apoptosis seems to be a main function of these receptors. It is astonishing that a member of this family, CD30, is overexpressed by highly proliferating tumours such as Hodgkin lymphoma (HL) and anaplastic large cell lymphoma (ALCL). It is known that CD30 stimulation leads to the apoptosis of ALCL tumour cells but not of Hodgkin-Reed-Sternberg (HRS) cells. We have already established the overexpression of TRAF1 in HRS cells. In this study we demonstrate that A20 is highly expressed in the HRS cells in 20/22 of cases of classical HL, in 4/4 cases of nodular lymphocyte-predominant HL (NLPHL), and in 2/2 cases of the anaplastic variant of diffuse large B cell lymphoma. In contrast, all other non-Hodgkin lymphomas, including ALCL, revealed either no A20 reactivity, or reactivity in less than 1% of all tumour cells. CD30 stimulation induced A20 and TRAF1 expression. This effect was most prominent in HL and ALCL cell lines with low basal expression levels of these molecules. Immunohistological studies of reactive lymphoid blasts in tonsillar tissue demonstrated that co-expression of CD30, A20, and TRAF1 also occurs in vivo. Cell lines with high basal A20 and TRAF1 expression were resistant to CD30-mediated apoptosis. The sensitivity to CD30-induced apoptosis was increased by inhibition of protein synthesis. TRAF1 transfection decreased CD30-induced apoptosis. Our data suggest that A20 and TRAF1 contribute to apoptosis resistance and, therefore, play an important role in the pathogenesis of classical HL. PMID- 12754743 TI - Beta-catenin mutations in pulmonary blastomas: association with morule formation. AB - To elucidate the contribution of beta-catenin gene mutation to the development of pulmonary blastomas, we analysed mutations in three well-differentiated fetal adenocarcinomas (WDFAs) and six biphasic pulmonary blastomas (BPBs). For comparison, eight clear-cell adenocarcinomas with fetal lung features were also examined. beta-Catenin gene mutations were found in all three WDFAs, two BPBs, and none of the clear-cell adenocarcinomas with fetal lung features. All tumours with mutations had a common histological feature, namely morule formation, and showed a characteristic heterogeneous beta-catenin expression pattern that was revealed by immunohistochemistry. Strong nuclear/cytoplasmic expression of beta catenin was seen in clustered cells in the morular areas and in single cells in glands, and was associated with neuroendocrine differentiation. As beta-catenin mutations are rare among lung tumours, this distinctive genetic feature, which is also immunohistochemically detectable as overexpression with a heterogeneous pattern, has diagnostic significance. The presence of this common genetic alteration found in both WDFA and BPB implies a histogenetic linkage between these tumours. PMID- 12754744 TI - DEC1 (STRA13) protein expression relates to hypoxia- inducible factor 1-alpha and carbonic anhydrase-9 overexpression in non-small cell lung cancer. AB - Differentiated embryo-chondrocyte expressed gene 1 (DEC1) is involved in cell differentiation, proliferation, and apoptosis, and was recently shown to be regulated by hypoxia. The present immunohistochemical study demonstrates extensive nuclear expression of the protein in 38% of a series of 115 non-small cell lung carcinomas using a polyclonal antibody (Ab) recognizing DEC1 protein. Such expression was directly related to the expression of two hypoxia-regulated proteins, namely the hypoxia-inducible factor (HIF) 1alpha and carbonic anhydrase 9. Although DEC1 was not related to angiogenesis or to the expression of VEGF and thymidine phosphorylase, a direct association with up-regulated bFGF receptors was noted. DEC1 was persistently expressed in the nuclei of normal bronchial and alveolar tissue. It is suggested that loss of DEC1 expression is an early event in the development of lung cancer, while DEC1 gene expression occurs in a subset of tumours and parallels the overexpression of other hypoxia-regulated proteins. PMID- 12754745 TI - BCL-2 family proteins in peripheral T-cell lymphomas: correlation with tumour apoptosis and proliferation. AB - The present study investigated expression levels of the anti-apoptotic proteins BCL-2, BCL-XL and MCL-1 and the pro-apoptotic proteins BAX and BCL-XS in a series of 112 peripheral T-cell lymphomas (PTCLs) classified according to the WHO classification. Using immunohistochemical methods and a 10% cut-off, each protein was detected in a subset of PTCLs: BCL-2 in 46%, BCL-XS in 49%, BAX in 57%, BCL XL in 57%, and MCL-1 in 65%. The mean percentage of positive cells for these proteins varied significantly among the PTCL types. Only two types of PTCL, ALK positive anaplastic large cell lymphoma (ALCL) and enteropathy-type T-cell lymphoma, had a distinctive pattern of expression; all were BCL-2-negative and MCL-1-positive. The mean percentage of BAX-positive and BCL-XS-positive tumour cells was higher in ALK-positive ALCL than in ALK-negative ALCL or other types of PTCL (p = 0.06 and p = 0.01, respectively, Kruskal-Wallis test). MCL-1 was detected significantly more frequently (p = 0.01, chi-square test) and at higher levels (p = 0.0001, Kruskal-Wallis test) in ALK-positive ALCL and ALK-negative ALCL than in other PTCL types. The apoptotic rate, evaluated by the TUNEL assay, correlated inversely with BCL-2 expression (p = 0.035). The proliferation index, assessed by the MIB-1 antibody, correlated with expression levels of MCL-1 (R = 0.42, p = 0.003), BCL-2 (R = 0.32, p = 0.027), BAX (R = 0.33, p = 0.014), and BCL XL (R = 0.34, p = 0.015) (Spearman rank). In conclusion, BCL-2 family proteins are expressed by a subset of PTCLs and their levels correlate with some histological types, apoptotic rate, and proliferation index. Expression of these proteins may explain the poor response of many types of PTCL to standard chemotherapy. These proteins may also provide novel targets for experimental therapy. PMID- 12754746 TI - Expression of apoptosis regulators in cutaneous T-cell lymphoma (CTCL) cells. AB - This study examined cutaneous T-cell lymphoma (CTCL) cell lines and cutaneous lesions for the presence of Bcl-2 gene family members and found that the two apoptosis-inhibiting members Bcl-xL and Mcl-1 and the two apoptosis-supporting members Bad and Bax were expressed. However, Bad was at least partially inactivated by phosphorylation. In skin lesions, the translocation of Bad from the nucleus to the cytoplasm may reflect the Bad inactivation by phosphorylation identified in vivo. Bax is also ineffective, as the non-steroidal anti inflammatory drug sulindac, whose cytotoxic effect is mediated by Bax, could not induce apoptosis in CTCL cell lines. The expression of Bcl-2, Bcl-xL, and Mcl-1 may therefore be sufficient to guarantee the survival of malignant CTCL cells. The Bcl-x, Mcl-1, Bad, and Bax proteins were also expressed in all CTCL skin lesions tested. In two patients from whom two biopsies from two different time points of the disease were available, a significant increase in Mcl-1 expression was found in the later-stage skin lesion. Overexpression of Mcl-1 and synthesis of non-functional Bax may be responsible for the resistance of CTCL cells to the anti-cancer drugs chlorambucil and sulindac. PMID- 12754747 TI - Recruitment of immature plasmacytoid dendritic cells (plasmacytoid monocytes) and myeloid dendritic cells in primary cutaneous melanomas. AB - The present study has analysed the distribution and phenotype of dendritic cells (DCs) in primary cutaneous melanomas and sentinel lymph nodes by immunohistochemistry. In primary melanomas, an increase of DCs was found in the epidermis and the peritumoural area. Intraepidermal DCs were mostly CD1a(+)/Langerin(+) Langerhans cells. Peritumoural DCs included a large population of DC-SIGN(+)/mannose-receptor(+)/CD1a(-) DCs, a small subset of CD1a(+) DCs, and, remarkably, plasmacytoid monocytes/plasmacytoid DCs (PM/PDCs). The PM/PDCs, most likely recruited by SDF-1 secreted by melanoma cells, produced type I interferon (IFN-I), but the expression of the IFN-alpha inducible protein MxA was extremely variable and very limited in the majority of cases. All DC subsets were predominantly immature. The peritumoural area also contained a minor subset of mature CD1a(+) DCs. However, the small amount of local interleukin (IL) 12 p40 mRNA and the naive phenotype of 20-50% of peritumoural T-lymphocytes are consistent with poor T-cell stimulation or erroneous recruitment. In sentinel lymph nodes, notable expansion of mature CD1a(+)/Langerin(+) DCs was observed. The paucity of intratumoural DCs and the predominant immature phenotype of peritumoural dermal DCs indicate defective maturation of primary cutaneous melanoma-associated DCs, resulting in lack of T-cell priming. These results may explain why melanoma cells grow despite the presence of infiltrating immune cells. PMID- 12754750 TI - Re: Scott. Radiotherapy and rectal cancer. J Pathol 2002; 197: 4-5. PMID- 12754748 TI - Re: Sastre-Garau et al. Inflammatory myofibroblastic tumour (inflammatory pseudotumour) of the breast: clinicopathological and genetic analysis of a case with evidence for clonality. J Pathol 2002; 196: 97-102. PMID- 12754751 TI - Labels issued for spermicide. PMID- 12754752 TI - AIDS cases on the rise. PMID- 12754754 TI - Syphilis is back. PMID- 12754753 TI - Internet may be fueling HIV infections. PMID- 12754755 TI - Integrate benchmarking with your PI efforts for long-term success. AB - Persistence and consistency are hallmarks of a strong program. Consider benchmarking just one of many performance improvement tools. Tools, structure, and accountability form the foundation of benchmarking. PMID- 12754756 TI - Closed-circuit TV wins fans in children's hospital. AB - Programming eases sense of isolation, promotes family bonding. Patients can watch in rooms, clinics, emergency department, and intensive care unit. Child Life specialists help normalize stay, relieve pre-procedure anxiety. PMID- 12754757 TI - Alliance will evaluate clinical excellence. AB - Intent is to provide organizationwide, measurable goals for hospitals. All 5,000 hospitals in the United States are being rated on clinical outcomes. Evaluations will examine 20 procedures in six different specialty areas. PMID- 12754758 TI - Leapfrog finalizing new incentive plans. AB - Incentives to be realigned for facilities meeting recommendations. Each practice brings different potential savings and costs, based on payer mix. Leapfrog will provide hospitals with a formula for calculating savings. PMID- 12754759 TI - Simple 'PUSH' spells improved senior health. AB - Seniors can achieve significant gains in strength and balance in weeks. Even the researchers find the results surprising. Study finds connections between mental state and physical performance. PMID- 12754760 TI - WHO extends its SARS-related travel advice to Beijing and Shanxi Province (China) and to Toronto (Canada), 23 April 2003. PMID- 12754761 TI - Progress towards global eradication of poliomyelitis, 2002. PMID- 12754762 TI - Angels or aliens? Refugee nurses in Britain, 1938 to 1942. PMID- 12754764 TI - Medicine, health and economic development: promoting spa and seaside resorts in Scotland C. 1750-1830. PMID- 12754763 TI - A shadow of orthodoxy? An epistemology of British hydropathy, 1840-1858. PMID- 12754766 TI - Illustrations from the Wellcome Library. Joseph Fenton and his books. PMID- 12754765 TI - Dangerous yardstick? Early cost estimates and the politics of financial management in the first decade of the National Health Service. PMID- 12754767 TI - The territory between life and death. Essay review. PMID- 12754768 TI - Performance and accountability in anesthesia. PMID- 12754769 TI - In-hospital resuscitation: can we do better? PMID- 12754770 TI - Invasive monitoring: what does the future hold? PMID- 12754771 TI - Pediatric anesthesia--a review of current progress. PMID- 12754772 TI - Can anesthesia improve the outcome in high risk obstertric patients? AB - Obstetric anesthetists should have an infrastructure that allows for referral and assessment of high risk patients. Management plans should be agreed well before delivery. This information must be available to other members of the team. Protocol for common high risk problems should be agreed and introduced. Promoting regional blockade for Cesarean section will reduce maternal anesthetic mortality. Epidural anesthesia preserves fetal biochemistry better than other forms of anesthesia. PMID- 12754773 TI - Ischemic pre-conditioning: helping or hindering the natural protection? PMID- 12754775 TI - Combined spinal/epidural anesthesia. PMID- 12754776 TI - Improving pain management. PMID- 12754774 TI - Controversies in critical care management of neurosurgical patients. PMID- 12754777 TI - Neuropathic pain and sympathetic nerve blocks. PMID- 12754778 TI - A prospective study of incidence of postoperative nausea and vomiting in a tertiary care hospital in Oman. AB - INTRODUCTION: This is a pilot study conducted in a tertiary referral center in Oman, to assess the incidence of postoperative nausea and vomiting (PONV) so that an institutional policy towards its alleviation could be evolved. METHODS AND MATERIALS: This is a prospective study of 491 patients undergoing obstetric and gynecological, general surgical, pediatric and urological surgical procedures that received general anesthesia, regional anesthesia or a combination of both. Data regarding the incidence of PONV was collected and was documented in a standardized questionnaire based on the patient history, anesthetic protocol, a postoperative interview of the patient and a review of the nursing records. RESULT: The over all incidence of PONV in our institution was 19.6%. The occurrence of vomiting compared to nausea or retching was high (p < 0.01). PONV was less in urological cases (p < 0.05) compared to general surgical cases. The incidence of PONV was similar (p > 0.05) in both groups, whether the patients received anti emetics along with pre-medication or not. PONV occurrence was significant in the first 6 hrs postoperatively (p < 0.01). In those patients who received propofol the prevalence of PONV was significantly less (p < 0.01). CONCLUSIONS: There should be a reassessment of the existing anti emetics in the institution. Propofol was found to be a good anti emetic. PMID- 12754779 TI - Keeping up with the literature: strategies for success. PMID- 12754780 TI - [Conducting a systematic review of the literature: a brief guide]. PMID- 12754781 TI - Living with ovarian cancer: women's perspectives on treatment and treatment decision-making. AB - In the year 2002, 2,500 women will have been diagnosed with ovarian cancer. Treatment for ovarian cancer is arduous, involving invasive surgery, chemotherapy, and/or radiation therapy. Studies have described the side effects of ovarian cancer treatment, but little has been written about women's perspectives on receiving that treatment. This work was undertaken to describe women's perceptions of living with ovarian cancer and their experiences with treatment. Eighteen women who had been diagnosed with ovarian cancer were interviewed in depth and theme analysis was undertaken with their transcripts. Three prominent themes emerged during the analysis: 1) initial treatment plans were overwhelming, 2) involvement in treatment decision-making was minimal, and 3) treatment had many side effects and complications. PMID- 12754782 TI - Finding and interpreting cancer screening guidelines: an oncology nursing challenge. AB - It is our hope that increased awareness of cancer screening recommendations will initiate a dialogue between individuals and their health care providers. Organizational screening recommendations differ as a result of evolving technology, financial and ethical considerations, and impact on mortality. It is through discussions with informed health care providers that patients and families will determine the most effective personal cancer screening strategies. We are hopeful that this discussion of cancer screening recommendations will encourage other oncology nurses to identify their provincial screening guidelines and to address them in discussions with people living with cancer, family members, community groups, and friendship circles. For those cancers where early detection correlates with improved survival, regular screening may have a direct impact on individual lives. We challenge you to implement strategies to incorporate cancer screening information in your professional and personal relationships. PMID- 12754783 TI - Cancer nursing in Ontario: defining nursing roles. AB - The delivery of cancer care in Ontario is facing unprecedented challenges. Shortages in nursing, as in all professional disciplines, are having an impact on the delivery of cancer care. Oncology nurses have a major role to play in the delivery of optimum cancer care. Oncology nursing, when adequately defined and supported, can benefit the cancer delivery system, patients, and families. A primary nursing model is seen as being key to the delivery of optimum cancer care. Primary nursing as a philosophy facilitates continuity of care, coordination of a patient's care plan, and a meaningful ongoing relationship with the patient and his/her family. Primary nursing, when delivered in the collaboration of a nurse-physician team, allows for medical resources to be used appropriately. Defined roles enable nurses to manage patients within their scope of practice in collaboration with physicians. Enacting other nursing roles, such as nurse practitioners and advanced practice nurses, can also enable the health care system to manage a broader number of patients with more complex needs. This article presents a position paper originally written as the basis for an advocacy and education initiative in Ontario. It is shared in anticipation that the work may be useful to oncology nurses in other jurisdictions in their efforts to advance oncology nursing and improvement of patient care. PMID- 12754784 TI - Sibling adaptation to the family crisis of childhood cancer. PMID- 12754785 TI - PFS and outpatient therapy: managing minutes and edits as a team. PMID- 12754786 TI - Countdown to HIPAA security, transactions standards compliance. PMID- 12754788 TI - [Recretion of enzymes and hormones by exocrine glands]. AB - The article reviews a poorly explored issue of secretive physiology-recretion from blood by glandulocites of various endocrinal glands of hydrolytic ferments and hormones that have been synthesized by digestive and endocrinal glands. The article discusses potential physiological role of the recretion function and the diagnostic significance of information obtained from analysis of recreted ferments and hormones in exosecretions. PMID- 12754787 TI - [Leptin--a new hormone in endocrinology]. AB - Research of recent years has fundamentally revised modern endocrinology. Many organs and tissue that have never before been treated as endocrinal or involved in production of various hormones, became such. In particular, adipose tissue secreting to blood an important hormone--leptin--became the study object of particular interest. PMID- 12754789 TI - [The involvement of nitric oxide in formation of hemoglobin oxygen-binding properties]. AB - The analysis of literature and results of our investigations indicate the possible involvement of L-arginine-nitric oxide (NO) system in formation of blood oxygen-carrying capacity. In reaction with hemoglobin NO forms methemoglobin, nitrosyl-hemoglobin (HbFe2+NO) and S-nitrosohemoglobin (SNO-Hb). The NO hemoglobin derivatives have the various biological functions (NO transport, storage, elimination etc.) and are involved in the genesis of different pathologic conditions. The presence of different NO-hemoglobin derivatives can differently influence on the whole blood hemoglobin-oxygen affinity (HOA): methemoglobin and SNO-Hb increases, and HbFe2+NO decreases it. Their effect on the blood oxygen-binding properties may be important for the gas exchange processes. At the level of lung capillaries such effect may be the additional mechanism promoting a blood oxygenation, and in the systemic microcirculation it may optimize blood desaturation and hence the tissue oxygen delivery. Blood oxygen-binding properties affect the state of L-arginine-NO system, however this system also may determine HOA through the intraerythrocytic regulatory mechanisms, oxygen-dependent nature of NO generation, regulation of vascular tone and effect of peroxynitrite. PMID- 12754790 TI - [Morphological analysis of information processing in basal ganglia of mammals]. AB - The article reviews published and own data on special features of organization of afferent cortical and infracortical correctional systems of the total striopallidaric nuclear complex in various species of mammals. The article focuses on analysis of organization of these flows in a dog being a classical object of behavioural experiments and a rare object of neuromorphological studies. It was discovered that the general trend towards segregated and converged flows of functionally different information at the level of striopallidum is retained in all tested mammals, being an evidence of the fundamental nature of the discussed principles of organization of the projection systems of basal ganglia. The reviewed morphological data confirm reliability of the presented concept of basal ganglia functioning. PMID- 12754791 TI - [Structural organization of rabbit neocortex in prenatal ontogenesis]. AB - In the review on the basis of data of different authors, the development dynamics of the rabbit neocortex in prenatal life is presented. The main points of neurons development, interneuronal contacts, layer-by-layer and areal differentiation as well as neocortex afferental and efferental communications, that are elucidated ambiguously by investigators, are discussed. For the purpose of determination of the neocortex maturation to the birth-date the comparative analysis of the reference morphological data and author's own neurophysiological data is accomplished. PMID- 12754792 TI - [The functional modifications under the long-term adaptation to cold]. AB - The data on modifications and interrelation of the afferent and efferent parts of the thermoregulatory system under the long-term adaptation to cold are presented. The body of data evidences for the important role of the thermoreceptors in the adaptive changes of the cold-defense responses. PMID- 12754794 TI - [Structural and functional changing induced by exposure to adaptive doses of X rays in the human lymphocytes both normal and defective by reparation of DNA double strands breaks]. AB - In the present work it is shown that the phenomenon of interphase chromosome centromeric region displacement, earlier revealed by the authors, is not realized in G0-lymphocytes with heterozygous BRCA1/2 gene mutations. The role of these genes in DNA double strand break (DSB) reparation is known. It is concluded that chromosome locus displacement is necessary for DSB repair, at least in the process of homologous recombination. In accordance with our data, some feature (pericentromeric cluster disintegration and displacement, the nucleus size increasing) characteristic for S- and G0-lymphocytes are observed in normal G0 lymphocytes treated with 3 and 10 cGy. However, the size of nucleus in G0 lymphocytes is restored through 6 hours after irradiation in opposite to the process in dividing cells. It was proposed that some typical for resting cell functions of G0-lymphocytes after inducing by adaptive doze of radiation are stopped as similarly as after stimulation of cells. Interestingly, that the process of the induced chromosome loci displacement is correlated with the decreasing of DNA reparation possibilities under UV-irradiation. The induced apoptosis level also decreases when chromosome loci are displaced. The possible mechanisms of the revealed phenomenon are discussed. This research supported by RFBR grant (No. 01-04-49180). PMID- 12754796 TI - [Effect of hydrogen peroxide in low and superlow concentrations on DNA structure]. AB - Hydrogen peroxide in low concentrations have effect on DNA structural characteristics both in solution at 38 degrees C and in vivo, in mice organs. PMID- 12754795 TI - [Ultraweak thermalized neutron field as an underestimated factor of DNA structural transformation]. AB - It was shown that thermalized neutrons in superlow doses were capable of inducing detectable structural transformations in DNA films and DNA solutions. PMID- 12754797 TI - [Polygenomic realization of mutagenic effects in the body of people exposed to low-dose radiation]. AB - A concept of polygenomic realization of mutagenic effects in the human body exposed to low-dose radiation on the basis of cell reproductions of genomic damages (Nig = 2n/2) was suggested. The above-said is in agreement with the principles of non-threshold mutagenic action of radiation and biological amplifier, and lead to the development of polygenomic dysbalance with pathophysiological consequences. PMID- 12754798 TI - [Low doses of radiation decrease the level of spontaneous and gamma-induced chromosomal mutagenesis in bone marrow cells of mice in vivo]. AB - Low doses of ionizing radiation are known to induce adaptive response (AR), which is characterized in most cases by temporary nature, though the possibility of long-term persistence of AR is not ruled out. In this investigation we studied the effect of low doses of gamma-radiation on both high-dose radiation-induced and spontaneous level of cytogenetic damage throughout the life of mice. SHK male mice 2 months old were used. Priming doses of 0.1 and 0.2 Gy (0.125 Gy/min, gamma radiation from 60Co) were used. A challenging dose of 1.5 Gy (1 Gy/min) was used in the experiments using a routine AR experimental design. The frequency of micronucleated polychromatic erythrocytes in bone marrow cells of primed, primed and challenged, and control groups was assessed at various times of animal life span. It was shown that: a) single low-dose gamma-irradiation induces a cytogenetic AR which can be revealed at 1, 3, 6, 9, 12 months after priming; b) single low-dose gamma-irradiation decreases the cytogenetic damage to a level below the spontaneous rate at the end of lifetime (20 months) of animals; c) ability to induce adaptive response does not depend on the age of animals at the moment of priming irradiation. In conclusion, the mechanisms underlying AR not only protect from chromosome damage induced by high-dose irradiation but also may play a role in spontaneous mutagenesis during aging of animals. PMID- 12754799 TI - [Estimation of genetic effects of chronic exposure to low-dose rate gamma radiation by cytogenetic methods and DNA-comet assay]. AB - The study of genetic effects in CBA/lac mice exposed for 1 year to constant low dose-rate gamma-radiation at a dose-rate 63 cGy/year has been carried out. We have shown the significant increase in the DNA breaks' level in spleen lymphocytes by comet-assay beginning from the total absorbed dose of 20 cGy. It is possible that the DNA breaks' level increase resulted from the structural rearrangement of chromatin or induction of lymphocyte proliferation. The results obtained by micronucleus test have proved that the mutagenic effect of chronic low dose-rate gamma-radiation depends on cell type and respectively on cell proliferation rate, cell differentiation, etc. So, by the end of experiment the significant increase in the frequency of PCE with micronuclei (MN) was observed. However, in contrast, the frequency of NCE with MN was not increased. No significant increase in the percent of lung cells with MN was registered. PMID- 12754800 TI - [The reaction of cell population to low level of irradiation]. AB - The results of long-term investigations of the effectivity of low level irradiation (below 0.5 Gy) carried out on the cells in culture and blood human lymphocytes (adults and children) have been brought. In the experiments conducted in the laboratory conditions and in the contaminated with radionuclides regions (after Chernobyl accident) the genomic instability have been discovered. The cell manifestations of the genomic instability have been registered in the progeny of irradiated cells as the decreasing of proliferative activity, the increasing of the frequency of cells with micronuclei, the increasing of cells with sister chromatid exchanges, the late cell death, the absence of the adaptive response ability, the enhancement of the radiosensitivity. The results of the investigations of the adaptive response of blood lymphocytes have been presented. It was shown that in all populations investigated there are individuals without the adaptive response and the individuals with the enhancement of radiosensitivity after adaptive irradiation (0.05 Gy). On the basis of own results and the data of literature the possible mechanisms of low level irradiation effects are discussed. The conclusion is that: a. The population with new properties can be formed after low level irradiation; b. The effects and mechanisms of this effect realization can be different from that after irradiation with high doses. PMID- 12754801 TI - Cell-density dependent effects of low-dose ionizing radiation on E. coli cells. AB - The changes in genome conformational state (GCS) induced by low-dose ionizing radiation in E. coli cells were measured by the method of anomalous viscosity time dependence (AVTD) in cellular lysates. Effects of X-rays at doses 0.1 cGy--1 Gy depended on post-irradiation time. Significant relaxation of DNA loops followed by a decrease in AVTD. The time of maximum relaxation was between 5-80 min depending on the dose of irradiation. U-shaped dose response was observed with increase of AVTD in the range of 0.1-4 Gy and decrease in AVTD at higher doses. No such increase in AVTD was seen upon irradiation of cells at the beginning of cell lysis while the AVTD decrease was the same. Significant differences in the effects of X-rays and gamma-rays at the same doses were observed suggesting a strong dependence of low-dose effects on LET. Effects of 0.01 cGy gamma-rays were studied at different cell densities during irradiation. We show that the radiation-induced changes in GCS lasted longer at higher cell density as compared to lower cell density. Only small amount of cells were hit at this dose and the data suggest cell-to-cell communication in response to low-dose ionizing radiation. This prolonged effect was also observed when cells were irradiated at high cell density and diluted to low cell density immediately after irradiation. These data suggest that cell-to-cell communication occur during irradiation or within 3 min post-irradiation. The cell-density dependent response to low-dose ionizing radiation was compared with previously reported data on exposure of E. coli cells to electromagnetic fields of extremely low frequency and extremely high frequency (millimeter waves). The body of our data show that cells can communicate in response to electromagnetic fields and ionizing radiation, presumably by reemission of secondary photons in infrared submillimeter frequency range. PMID- 12754802 TI - [Influence of chronic exposure to low doses of space ionizing radiation on the character of formation of microbial assemblage in the habitat of orbital station]. AB - Statistically valid relations between radiation conditions in compartments of MIR station and the micromicete population (CFU number) on the surface of the equipment and the interior have been established. It was found that in conditions of a chronic exposure to space radiation the number of CFU increased in one thousand and more times with increasing of absorbed dose rate from 200 up to 1000 microGy/day. The results of land-based model experiments confirmed morphological changes in the "flight" strains of funguses under exposure to low doses of gamma (100-800 microGy/day) and neutron (0.2-2 neutron/cm2.s) radiation. It was found that the morphological changes in the control (museum) cultures of funguses of the same species, which were expressed in the weak increase of vegetative mycelium, were detected only after repeated gamma- and gamma + neutron irradiation. PMID- 12754803 TI - [Some effects of radiation hormesis for bacterial and yeast cells]. AB - A comparative study of chronic and acute action of ionizing radiation on the processes of aging and dying off of bacterial and yeast cells was carried out. It was ascertained that chronic action of ionizing radiation, 2-10,000 times exceeded the natural background, resulted in slowing down of aging and dying off of both pro- and eukaryotic cells. A single acute irradiation of yeast also resulted in the retardation of dying off of the yeast cells surviving after irradiation. The data is presented demonstrating a great increase in the survival of yeast cells under their repeated irradiation after recovery from potentially lethal radiation. PMID- 12754804 TI - [Non-monotonous dose-response relationship in the region of low doses of ionizing radiation]. AB - The statement about nonmonotony of dose-effect curves as a result of nonmonotony of the time-effect relationship including the field of low doses is discussed. The living cells possess a fundamental property to response to action of different stress agents by oscillatory--nonmonotonous--hanges of metabolism. The systems keeping up homeostasis by direct and feed-back regulation return metabolism to norm. In the fixed temporary point a dose-effect dependence may take the nonmonotonous character e.g. reverse dose-response relationship. The changes of the oscillation parameters suggested the inclusion of the different pathway for homeostasis keeping. Radiation hormesis does not focused on the metabolic and functional nonmonotonous response. Radiation stimulation is considered as consequence of the peculiarity of the homeostasis maintenance pathways in the certain interval of the low doses of ionizing radiation. PMID- 12754805 TI - [Small doses of X-ray irradiation activate the NO-synthase component of the nitric oxide cycle]. AB - It has been shown that chronic X-ray irradiation, (CRI), activates the formation of NO in rats. This is apparent in the increase in the level of NO2- in the blood plasma from 12.59 +/- 1.7 to 39.79 +/- 2.9 nmol/ml after 10 days of irradiation. On the 20 and 30 day of CRI, the level of NO2(-)- was 21.05 +/- 1.2 and 30.73 +/- 1.9 nmol/ml respectively. The changes in the NO-synthase component of the NO cycle were accompanied by a decrease in the osmotic resistance of the erythrocytes and the nitritreductase activity of hemoglobin. PMID- 12754806 TI - [Influence of low doses of ionizing radiation on tenascin expression in hybridoma cell systems]. AB - One of the achievements of the modern radiation ecology is the preparation and application of stable eukariotic cell lines to solve various problems occurring under exposure to ionizing radiation, especially to low doses. The detection of onco-fetal protein--tenascin in different embryonic and tumor cells of humans and animals supposes the probability of appropriate gene expression in lymphoid cells, including hybridomal cells. Using the immunochemical method, the study of tenascin expression in two mouse hybridomal lines was carried out. Tenascin was revealed in hybridomal lines MLC-1 and K-48. Further hybridomal cell lines were exposed to X-ray radiation (120 KV) with doses 2.10,15 cGy. The obtained results demonstrated the sensitivity of tenascin expression to low doses of ionizing radiation, that may be used as a convenient model of studying of genotoxic effects of various damaging ionizing agents on a cell level. PMID- 12754807 TI - [The effect of low doses of radiation on the glutathione status in children and adults after the Chernobyl Atomic Electric Power Station accident]. AB - A level of glutathione in the blood plasma of human population exposed to low dose radiation after the Chernobyl accident was studied; a complicated character of dose dependence was shown. The response of the glutathione system to low and high radiation doses is different in the children living in the radionuclide contaminated areas and in the participants of the elimination works. PMID- 12754809 TI - Strontium fallout from Chernobyl and perinatal mortality in Ukraine and Belarus. AB - Perinatal mortality rates in the regions of Ukraine and Belarus surrounding the Chernobyl site increased in 1987, the year following the Chernobyl accident. The same year, increases of perinatal mortality were also observed in Germany and Poland, and the effect can be associated with the caesium burden in pregnant women. After 1989, there is an unexpected second rise of perinatal mortality in Belarus and Ukraine. This increase is shown to correlate with the strontium content in pregnant women. The findings parallel an increase of perinatal mortality in Germany following the atmospheric bomb tests in the 1950's and 1960's. While the effect from caesium is essentially limited to 1987, the effect from strontium persists until the end of the study period in 1998. The cumulative effect from strontium around Chernobyl outweighs the effect from caesium by at least a factor of 10. This is contrary to the assertion that the caesium content in the Chernobyl fallout was more than 10-times greater than the strontium content. Thus, the dose factor presently used seems to severely underestimate the effect of strontium on perinatal mortality. PMID- 12754808 TI - [High radiopathogenic children's loads caused by "low doses" of technogenic chronic radiation]. AB - To study influence of "low radiodoses" on their early effects, the method to monitor peculiarities for human biochemphysic pathologies caused by technogenic chronic doses divided by natural constant radiation background doses (relative radiation loads as their ratios, RRL) has been developed. High radiolabile coupled system of harmful catabolites of the initiated lipoperoxic cascade with essential radioprotectors (vitamins A and E as natural antioxidants) as pathogenic significant test-objects were monitored. For exposed children low radiation technogenic doses were high enough as RRL to disturb and to dysregulate both systems. For evaluation of individual radiosensitivity significances of age for children especially, as of the biochemical status for both systems are shown. The approach permits to investigate pathologies of studied systems as primary factors for mechanisms of related radiogenic known somatic health secondary consequences and to use it for some important practical applications. PMID- 12754810 TI - [Late consequences of action of low doses of ionizing radiation on human enzymatic antioxidant system]. AB - Age-related changes in the activity of superoxide dismutase and glutathione peroxidase in the blood of participants of the Chernobyl accident liquidation were studied. According to our findings the people under 30 years old are the most sensitive to irratiation. PMID- 12754811 TI - [Parameters of peroxide oxidation of lipids and the system of its regulation in the tissues of Ellobius talpinus Pall, living on the territory of the East Urals Radioactive Trace]. AB - The peroxide oxidation of lipids and the systems of its regulation was tested in Ellobius talpinus Pall, living on the territory of the East Urals Radioactive Trace with the 90Sr contamination density of 1000 Ci/km2. The degree of sensitivity of the examined tissues and organs was revealed, the most considerable parameters were defined for marking radiosensitivity of the taken species, as well as the complex of parameters, allowing classifying an animal as an inhabitant of the clean or the contaminated territory. The found changes are considered as compensatory mechanisms for forming of nonspecific resistance. PMID- 12754812 TI - [Adaptation processes in natural Drosophila populations in radiation contaminated Belarus regions before and after radiation exposure removal]. AB - We have shown that natural drosophila populations from the settlement Vetka of Gomel region with increased radiation background are more adapted to mutagenic effect of radiation than drosophila populations from Berezinsky reserve (the control). After the populations were placed into laboratory thermostat adaptation of Vetka population remained within 6-8 generations without irradiation. However the control population became more resistant too. So, the keeping of natural drosophila populations under laboratory conditions was a stress and led to unspecific adaptation the same as a low level of radiocontamination did. These facts should be considered in studying dynamics of the mutation level during radionuclide removal in animals caught in radiocontaminated regions and placed in vivaria conditions. PMID- 12754813 TI - [Effects of 10-day long exposure to gamma irradiation at low doses on bone marrow cells in mice]. AB - Effects of ten day long exposure to gamma-irradiation at low doses (mean dose rate of 1.5-2.0 m Gy/day, total dose of 15 m Gy) on hemopoietic (CFU-S) and stromal (CFU-F) progenitor cells from murine bone marrow were examined. The CFU-F content measured as in vitro fibroblastic colony number showed 1.5-4.5-fold increase. Additionally, the size of ectopic marrow transplants evaluated by counting myelokariocytes and CFU-S numbers also increased. No significant changes of CFU-S proliferation rate were found. PMID- 12754814 TI - [Increase of probability of cell loss in the endothelium of blood vessel capillaries in progeny of rats after a low-dose irradiation of one of the parents (electron microscopy study)]. AB - The total gamma-irradiation of Wistar rats at a dose of 0.25 Gy as well as at higher doses (0.5, 2 and 4.5 Gy) produces in the capillary endothelial cells of myocardium and lung a pronounced, dose-independent increase of the yield of necrotized cells. Similar changes were revealed in the animals, of which one of the parents (a male one day, a female seven days prior to copulation) was irradiated at doses of 0.25 and 0.5 Gy. This effect was observed in all studied descendants. The massive induction of the changes already by low radiation actions and their dose-independence allow considering the revealed effects as a manifestation of peculiar cellular reactions that presumably have epigenetic nature. PMID- 12754815 TI - [State of the reproductive system and liver in male rats after fractionated radiation exposure to low doses as well as in their progeny]. AB - Male rats were exposed to fractionated 0.1 Gy radiation in early onthogenesis (1 month). Essential changes in reproductive system and liver were found. First generation offspring of the exposed males and females showed moderate radiation changes more expressed in immature rats. PMID- 12754816 TI - Cell analysis with the new Leipzig high-energy ion nanoprobe. AB - The high-energy ion nanoprobe LIPSION at the University of Leipzig has been in operation since 1998. The ultrastable, 3.5 MV SINLETRON accelerator supplies the H+ or He+ ion beam. A magnetic scanning system moves the focused beam across the sample. At present, a resolution of 41 +/- 4 nm in the low current mode and 300 nm at 5 pA can be achieved. The experimental chamber is equipped with electron-, energy dispersive X-ray-, and particle detectors. They can be used simultaneously to analyse the sample by means of PIXE (particle induced X-ray emission), RBS (Rutherford backscattering), and in the case of thin sections or monolayer samples STIM (scanning transmission ion microscopy). A goniometer allows the application of channeling measurements in single crystals in combination with these methods. In contrast to previous publication describing microbeam facility at LIPSION, the current biomedical research has concentrated on microscopy and tomography on chondrocytes in pig cartilages and fixed single endothelial cells (HUVEC). For the irradiation of single living cells, an external beam facility with irradiation platform, fast beamgate and mini-Petri dishes is under construction. PMID- 12754817 TI - [Analysis of the epidemiological data concerning radiation carcinogenic effects and approaches to the low doses' upper limits determination in the aspect of a threshold of the unhealthy influences of ionizing radiation]. AB - The analysis of the epidemiological data regarding cancer mortality in cohorts of Japanese A-bomb survivors and Chermobyl liquidators exposed to different doses suggests that there are good reasons for recognizing the threshold of the radiocarcinogenic effect in the region of about 200 Gy (mSv). The analysis of solid cancer mortality in Japanese cohort, which exceeded the expected one in a dose diapason of 5-200 mSv, revealed a (quasi) plateau in a dose-effect curve and led to the conclusion that the nature of the overshoot is non-radiogenic. The analysis of supposedly dose dependent leucosis incidence in the limited low dose diapason in the Chernobyl cohort showed that the real coefficient of the excess absolute or relative radiation risk could not be received in the case because the larger part curve was placed under the control level. In supporting the principle of single hit in a cell nucleus as a base of microdosimetric determination of low radiation doses, the approach to objective delimitation between low, intermediate and high doses regions has been proposed. The low doses upper limit of sparse ionizing radiation for cell nucleus of 8 microns in diameter has been evaluated as 0.65 mGy. It can serve for evaluation of the dose rate threshold regarding the safe chronic radiation levels in the environment. PMID- 12754818 TI - [Spontaneous leukosis as a model for an investigation of low and very low physical and physico-chemical effects on oncological process]. AB - Kinetic investigation of spontaneous leukosis in AKR mice have been carried in connection with a number of indices: changes in the mass of principal organs of the immune systems (thymus, spleen, lymphatic nodes), liver, alterations of haematological data (the sum of leukocytes, the percentage composition of blood cells, the quantity of undifferentiated cells), changes of physico-chemical conditions in cells (NMR-investigation). The dynamics of some of these indices and also life-spans of animals with leukosis after irradiation with doses 1.2-2.4 cGy (dose-rate 0.6 cGy/day) have been investigated. The enhancement of the frequency of leukosis and shortening the average and maximum life-spans of irradiated mice has been found. PMID- 12754819 TI - [The role of reaper-dependent apoptosis in radiation-induced life-span alterations in Drosophila melanogaster]. AB - This is the result of studying on a problem of radio-induced ageing. From the point of view of radiation genetics it is perspective to investigate influence of low doze irradiation on individuals with the mutant phenotypes that allows to assume a role of separate genes and mechanisms controllable by them in determination of life span and ageing. The role of a reaper-dependent way of apoptosis regulation in the induced change of life span is shown. The assumption is put forward that apoptosis has the important role during ageing an animal organism. PMID- 12754820 TI - [The influence of geomagnetic field on blood leukocyte radiosensitivity and gender determination in humans]. AB - The gender and age features of the geomagnetic field (GMF) influence (K-indexes) on the leukocyte radiosensitivity (S) in human blood in case of normal and enhanced levels of metal-ecotoxicants (Pb, Cd, Hg, Mn, Zn, Cu) in blood was studied, as well as the impact of the GMF intensity alterations on the child gender determination. In the whole studied population (n = 244) it was observed a negative relationship between S- and K-indexes. It was shown that the most changes of blood leukocyte radiosensitivity depended on the GMF oscillations in men older than 45 years. In case of metal-ecotoxicant concentrations in blood near normal level there were observed significant relationships between S- and K indexes, which was infringed by more than 2-fold excess of these toxicant concentrations in blood. Retrospective analysis showed that in case of the GMF intensity enhancement in the conception moment the girls were born mostly, and in case of the lowering--the boys were. PMID- 12754821 TI - [Comparative study of radioprotective efficiency of various doses of cystamine dissolved in normal saline solution or distilled water]. AB - In experiments on mice it has been found that radioprotective effect of cystamin used in a doze 0.15 mg/kg reveals only at its dissolution in a physiological solution, and it is not found out at dissolution of a preparation in distilled water. PMID- 12754822 TI - [Nutritive fibers of animal origin. Collagen and its fractions as essential components of new and efficient food products]. AB - Data on the production of collagen and its fractions (which in many cases surpass the parent protein in functional properties), reported in the literature throughout the past two decades, are reviewed. The material presented elucidates the role played by collagen and its fractions in the development of new, ecologically pure foodstuffs containing nutritive fibers. As follows from the analysis of the data, collagen fractions with molecular weights in excess of 120 kDa offer the greatest promise for producing useful foodstuffs. PMID- 12754823 TI - [Effect of the degree of acetylation of chitosan on its enzymatic hydrolysis with the preparation Celloviridin G20x]. AB - The degree of acetylation exerted only insignificant effects on the enzymatic hydrolysis of chitosan, while affecting the composition of the resulting hydrolysates and their water solubility. Chitosan with various degrees of acetylation was produced by reacetylation of the original chitosan (the solvents, methanol and 2% acetic acid, were present at a ratio of 54:51 v/v; the amount of acetic anhydride was in the range 0.1-2.0 mmol per 1 g chitosan). Hydrolysis by the enzymatic preparation Celloviridin G20x was performed at the enzyme to substrate ratio of 1:400 in sodium-acetate buffer, pH 5.2 (55 degrees C) for 1 h. PMID- 12754824 TI - Synthesis of ethyl isovalerate using Rhizomucor miehei lipase: optimization. AB - Immobilized lipase from Rhizomucor miehei (Lipozyme IM-20) was used to catalyze the esterification reaction between isovaleric acid and ethanol to synthesize ethyl isovalerate in n-hexane. Response surface methodology based on five-level four-variable central composite rotatable design was employed to optimize four important reaction variables such as enzyme/substrate E/S ratio, substrate concentration, incubation time, and temperature affecting the synthesis of ethyl isovalerate. The optimum conditions predicted for achieving maximum ester yield (500 mM) are as follows: E/S ratio, 48.41 g/mol; substrate concentration, 1 M; reaction time, 60 h; temperature, 60 degrees C. The predicted value matched well with experimentally obtained value of 487 mM. PMID- 12754825 TI - [Mechanism of overproduction of secreted enzymes in the mycelial fungus Penicillium canescens]. AB - The fungus Penicillium canescens strain F178 (VKPM) and its niaD- mutant exhibited an increased capability of synthesizing extracellular enzymes beta galactosidase (70-80 U/ml) and xylanase (100 U/ml). The synthesis was induced by arabinose and its catabolite, arabitol. A deficiency in arabitol dehydrogenase, leading to arabitol accumulation in the cell, was detected in the chain of reactions of arabinose catabolism. The increased synthesis of beta-galactosidase and xylanase in P. canescens is accounted for by (1) cellular accumulation of the inducer (arabitol) at low concentrations of arabinose in the medium and (2) prevalence of induction over repression. PMID- 12754826 TI - [Isolation and characterization of humin-like substances produced by wood degrading fungi causing white rot]. AB - Three samples of high-molecular-weight humin-like substances were obtained by solid-phase cultivation of Coriolus hirsutus and/or Cerrena maxima on oat straw. The yield of humin-like substances amounted to 1.38-2.26% of the weight of the plant substrate consumed. These substances, produced both by individual and mixed cultures of the basidiomycetes, were shown to be similar in their structure and physicochemical properties. According to the data of IR and 13C-NMR spectroscopy, the substances contained aromatic fragments and were close to soil humic acids. Studies of the dynamics of laccase production suggested that the humin-like substances were produced bia direct degradation of lignin macromolecules with direct involvement of extracellular laccase. PMID- 12754827 TI - [Specific features of fermentation of D-xylose and D-glucose by xylose assimilating yeasts]. AB - The ability to assimilate D-glucose and D-xylose was studied in 21 yeast species of the following genera: Candida, Kluyveromyces, Pachysolen, Pichia, and Torulopsis. All the cultures fermented D-glucose with the formation of ethanol. During the assimilation of D-xylose, ethanol was produced by P. stipitis and C. shehatae, whereas xylitol was produced by C. didensiae, C. intermediae, C. parapsilosis, C. silvanorum, C. tropicalis, K. fragilis, K. marxianus, P. guillermondii, and T. molishiama. The yeast P. tannophilus produced comparable amounts of both alcohols. The possible use of xylose-assimilating yeasts for the production of xylitol and ethanol is discussed. PMID- 12754828 TI - [Effect of antibacterial factors and cell metabolism biostimulants on bacterial luminescence]. AB - Bioluminescence was used as an index of effects of a series of factors (differing in origin and structure) on cell metabolism. Luminescence of intact cells of Escherichia coli lum+ (a genetically modified strain) was measured on exposure to antibiotics, probiotics, phages, and biostimulators. The sensitivity of E. coli lum+ to antibiotics correlated with the antibiotic-induced luminescence quenching. Bioluminescence-based assessment of the antagonistic activity of the main probiotics (bificoll, bifidobacterin, acylact, colibacterin, and a composite probiotic preparation), each taken at a concentration of 1 dose/ml demonstrated that bacterial luminescence was inhibited by 75-99.9% (exposure time, 30 min). Bioluminescence changes reflected cell damage associated with phage infection. It was shown that bioluminescence stimulation could be used as an index of the effect of immunomodulators (olexin and vermin) on cell metabolism in bacterial cultures. PMID- 12754829 TI - [Specific toxic effects of 2,4,6-trinitrotoluene on Bacillus subtilis SK1]. AB - Using Bacillus subtilis SK1 as an example, it was demonstrated for the first time that 2,4,6-trinitrotoluene (TNT) transformation pathways change with TNT concentration. The growth of cultured B. subtilis SK1, delayed at 20 mg/l TNT (minimum toxic concentration), was resumed following TNT transformation. Aromatic amines were predominant metabolites detected in the culture medium at early stages of TNT transformation. The culture growth was completely inhibited by 200 mg/l TNT. As this took place, nitrites accumulated in the culture medium. PMID- 12754830 TI - [Use of bacteria of the genus Azotobacter for biodegradation of oil-contaminated soils]. AB - The rate of self-purification of oil-contaminated soil increases after introduction of bacteria of the genus Azotobacter. The bacteria can assimilate oil hydrocarbons as the sole source of carbon and energy, both in the presence of fixed nitrogen and during nitrogen fixation. The species Azotobacter chroococcum activates growth of hydrocarbon-oxidizing bacteria present in Devoroil. PMID- 12754831 TI - [Growth of bacteria degrading naphthalene and salicylate at low temperatures]. AB - A total of 58 bacterial strains degrading naphthalene and salicylate were isolated from soil samples polluted with oil products, collected in different regions of Russia during winter and summer. The isolates were assessed for their ability to grow at low temperatures (4, 8, and 15 degrees C); bacteria growing at 4 degrees C in the presence of naphthalene or salicylate accounted for 65% and 53%, respectively, of the strains isolated. The strains differed in the temperature dependence of their growth rates. It was demonstrated that the type of expression of Nah+ phenotype at low temperatures depended on the combination of the host bacterium and the plasmid. PMID- 12754832 TI - [Quantitative assessment of the biological activity of chemicals by soil microbial associations]. AB - Quantitative assessment, using three Pseudomonas sp. strains, of the activity of the microbial biocide Soncid 8101 demonstrated that the values of effective sublethal concentrations (L50) differed by 500% (because of individual variations in the sensitivity of the test strains). The spread of parameters of biocidal activity could be narrowed by using a mixture of microorganisms with high, medium, and weak resistance. A method for quantitative assessment of the activity of microbial biocides was proposed, based on the use of natural associations of soil bacteria. PMID- 12754833 TI - [Optimization of conditions for storage and cultivation of the fungus Claviceps sp.--a producer of the ergot alkaloid agroclavine]. AB - Conditions of agroclavine biosynthesis by the mutant Claviceps sp. strain s 106 were studied. The content of agroclavine was maximum (1.5-2 g/l) on days 15-16 of cultivation in the complex medium T25, containing sucrose, citric acid, and yeast extract. Agroclavine was the major component of the alkaloid fraction (90-95%). Storage of the culture at -70 degrees C in T25 supplemented by 7% glycerol provided a stable level of alkaloid formation. PMID- 12754834 TI - [Phytohormone interactions in the control of proton translocation activity of plant cell plasmalemma]. AB - The effects of phytohormones (abscisic acid, gibberelic acid, and jasmonic acid) and ambiol (a synthetic growth regulator) on processes of H+ transport across the plasmalemma were studied in membrane vesicles isolated from the parenchyma of potato (Solanum tuberosum L.) tubers. Phytohormones and ambiol were tested either individually or in combinations. Each of the substances tested changed the initial rate of H+ uptake by the vesicles. Two signaling substances added to the incubation medium simultaneously modified the activity of each other. It is suggested that the interaction of a signaling substance with components of the plasmalemma alters the responses of the membrane to other signaling molecules. PMID- 12754835 TI - [Free and bound amino acids during germination of maize seeds with implemented foreign prolamines]. AB - Wheat prolamines were preincorporated into the endosperm of maize seed, and concentrations of free and bound amino acids were measured in the endosperm and seedlings during seed germination. Incorporation of foreign prolamines lowered the decrease rate of free proline concentration in both the endosperm (sprouting and shoots) and the embryo (sprouting). The seedlings at the stage of shoots were characterized by higher concentrations of aspartic acid, glutamic acid, and tyrosine. At the stage of transition to autotrophic nutrition, the pool of amino acids that are physiologically important for the plant growth was greater in maize seedlings with incorporated wheat prolamines. PMID- 12754836 TI - [Changes in the composition of the essential oil in stored marjoram]. AB - The composition of volatile components of the essential oil of marjoram plants (Majorana hortensis M.) and its stability during storage were studied by capillary gas chromatography and gas chromatography/mass spectrometry. Storage in the dark for 1 year was associated with insignificant changes in the composition of the essential oil, and its organoleptic characteristics remained largely unaffected. Storage in the light produced considerable changes in the composition of the oil, due to chemical transformation of terpenoids. PMID- 12754837 TI - [Polymeric systems with antithrombin activity for thermally activated targeting]. AB - Copolymers of N,N-diethylacrylamide and N-acryloylphthalimide with lower critical solution temperature (LCST) were synthesized by radical copolymerization. Polymeric systems with antithrombin activity and LCST were prepared via a reaction of amino groups of hirudin with phthalimide groups of the copolymers. On increasing hirudin content, LCST of the polymeric systems increased. The antithrombin activity of polymeric systems obtained by hirudin immobilization on copolymer carriers was inversely related to the content of the copolymer, amounting to 6% of the activity of native hirudin. PMID- 12754838 TI - [Kinetics of chemical reactions for quality prediction of canned fish during storage]. AB - Changes in a wide range of quality characteristics of canned fish were studied during storage at different temperatures. A number of biochemical parameters were found, which undergo significant monotonic changes in the course of storage, correlating with organoleptic scores. It was demonstrated that simulation of thermal aging of canned fish, based on the laws of chemical kinetics, may be used for predicting quality changes and determining the shelf life. PMID- 12754839 TI - [Neurophysiological mechanisms of voluntary attention: a review]. AB - The review is focused on attention as behavior-controlling process. Neurophysiological, electrophysiological and neuropsychological studies of different brain structures during voluntary attention are analyzed. These data show that selective voluntary attention modulates activity of sensory specific cortical zones involved in relevant signal processing. Fronto-thalamic system consisting of prefrontal cortex and thalamic mediodorsal nuclei is shown to be main source of top-down selective modulation of voluntary attention. The review proposed the hypothetical model of selective cortical activity modulation during voluntary attention based upon the available data and evidence of own electroencephalographic studies. PMID- 12754840 TI - [Sleep EEG as a nonlinear dynamic process: a comparison of global correlation dimension of human EEG and measures of linear interdependence between channels]. AB - The goal of this work was to study (1) whether the estimation of correlation dimension (D2) using spatial embedding distinguishes between sleep stages and (2) whether information gained from the application of global D2 is redundant to measures of linear interdependence between channels. Twenty one-channel EEG segments of 12 healthy male subjects recorded during waking and sleep stages REM, I, II, and III-IV (according to the Rechtshaffen and Kales criteria) were analyzed with global (multichannel) D2, mean square correlation coefficients (MS) and proportion of variance accounted for by the first principal component (PC1). D2 was found to decrease progressively from stage I to stage III-IV with D2 values of waking and REM being close to those of stages I and II. MS and PC1 did not distinguish among sleep stages but yielded significant differences between waking and sleep. The results suggest that global D2 extracts information from human EEG. That sort of evidence cannot be obtained with measures of linear interdependence between channels. PMID- 12754841 TI - [Multiple parameter comparative EEG analysis in alcoholism and narcotic dependence]. AB - Multiparametric comparative analysis of spatial organization of EEG was carried out in 137 alcoholics and 131 heroin addicts. Common and different deviations from normal EEG (105 control subjects) were found. Global alterations of EEG spatial organization were observed in drug addicts (as compared to alcoholics). Such changes characterized increasing synchronizing effects of mesolimbic and brainstem structures on the brain cortex. The ethanol effects were more specific and asymmetric. Changes in EEG spectral-coherence characteristics were revealed in all frequency band, however, maximal changes took place in the high-frequency theta in drug addicts and in narrow-frequency alpha subranges in alcoholics. Different effects on the high-frequency EEG component (19.00-21.25 Hz) and information-energy index (coherence-to-spectral power ratio) suggest the difference influence of ethanol and heroin on emotional-motivational and cognitive processes as well as the level of consciousness. The obtained data on EEG discrimination of alcoholism and drug addiction (the inverse problem solution) on the basis of "specific" EEG patterns appear to have considerable promise in development of systems of occupational selection. PMID- 12754843 TI - [Enhancement of glutamate release in the extracellular space of nucleus accumbens during presentation of aversive or neutral stimuli instead of food reinforcement]. AB - By means of in vivo microdialysis combined with HPLC/EC analysis it was shown that presentation to a rat of an inedible object (a piece of rubber) or an aversive object (food of bitter taste) instead of expected food caused a marked increase in extracellular glutamate level in n. accumbens. In rats not expecting food reinforcement, extracellular glutamate remained unchanged during presentation of these objects. Our findings suggest that dissociation between the expected biological value of a presented object and its real significance may be an important determinant for glutamate release control in n. accumbens during food behavior. PMID- 12754842 TI - [Comparative analysis of haloperidol effects on passive avoidance retention in aggressive and submissive mice]. AB - The effects of haloperidol on retention of avoidance during its extinction in C57BL/6J mice were shown to depend on a behavioral stereotype (aggressive or submissive). In submissive mice, haloperidol (0.5 mg/kg) injected an hour before training stabilized retrieval of the conditioned reflex in repeated testings (up to 17 days) as compared to its fast extinction in control animals. In aggressive mice, on the contrary, haloperidol reduced the retention of the memory trace retrieval. It is suggested that divergent haloperidol effects on extinction of passive avoidance in mice with alternative behavioral strategies are determined by the features of organization of the mesolimbico-cortical dopaminergic system and emotional state, in particular, anxiety. PMID- 12754844 TI - [Behavioral and morphological asymmetry in brain weight selected mice]. AB - Mice selected for large and small brain weight preferred to move rightwards in two cognitive paradigms, food-motivated T-maze performance and extrapolation ability tests. The degree of asymmetry in choices of movement direction varied in selection generations and sometimes was accompanied by asymmetry in the weight of the left and right hippocampi and other forebrain structures and different numbers of right and left neocortical cells. PMID- 12754845 TI - [Mathematical simulation of induction of long-term depression in cerebellar Purkinje cells]. AB - Mechanisms of associative and homosynaptic long-term depression (LTD) in cerebellar Purkinje cells are discussed. The possibility of LTD induction related to a decrease in efficacy of AMPA receptors through either their dephosphorylation or phosphorylation is investigated by mathematical simulation. PMID- 12754846 TI - [Kainate receptor activation enhances both tonic and phasic GABA-ergic inhibition in CA1 hippocampal interneurons]. AB - Kainate receptor agonists are powerful convulsants and excitotoxins. It has been a lot of controversy around functions of these receptors in the brain. It is shown in this article that kainate enhances evoked GABAergic IPSC (phasic currents) in CA1 interneurons in concentration-dependent manner. The phenomenon is likely to be due to kainate-mediated lowering of the threshold for action potential generation in interneuron axons and increased number of terminals responding to the same stimulus strength. Kainate application also induced an enhancement in tonic GABAergic conductance. This phenomenon can be attributed to massive extracellular GABA accumulation caused by interneuron firing in the presence of kainate. Extracellular GABA also shunts synaptic currents by activating tonic conductance as well as desensitizing synaptic GABAA receptors. Thus, the enhancement of the evoked IPSCs by 1 microM kainate was complicated by early and transient decrease. The kainate receptor-mediated enhancement of GABAergic tonic and phasic signalling to interneurons can contribute to the depression of GABAergic transmission to pyramidal neurons. The consequence of this phenomenon may play a major role in the epileptogenic action of this agent. PMID- 12754847 TI - [Immunization of rats with bovine serum albumin conjugate with dopamine or 5 hydroxytryptamine prevents the development of experimental MPTP-induced depressive syndrome. An electrophysiological study]. AB - In electrophysiological experiments in Wistar rats it was shown that preventive immunization of animals with bovine serum albumin conjugated with dopamine or 5 hydroxytryptamine or with bovine serum albumin alone partly protects against the development of experimental MPTP-induced depressive syndrome. Signs of depressive like state such as decrease in REM-sleep latency and the development of epileptiform activity in caudate putamen were not observed in immunized animals, whereas REM-sleep proportion in the total sleep structure was increased. Changes in the spectral characteristics of brain electric activity and sleep structure during development of experimental MPTP-induced syndrome in immunized rats are antigen-specific and reflect the functional shifts in neurotransmitter systems, both in those which are the target of the immune action and those sensitive to alterations in the immune state of a body. PMID- 12754848 TI - [Effects of estrogens on learning of rats with chronic brain cholinergic deficit in Morris water maze]. AB - A chronic deprivation of brain cholinergic functions in rats caused by intracerebroventricular injection of neurotoxin AF64A increases the escape latency in Morris water maze test as compared to control sham-operated animals. Measurements and analysis of rat movement tracks using an original computerized "Behavioral Vision" system revealed the ability of 17 beta-Estradiol and its synthetic isomer J-861 (both administered daily in per os dose 0.2 mg/kg during 7 days before and 10 days after a single intracerebroventricular injection of AF64A) to improve learning of the animals. Directivity of search trajectories was estimated by a novel index of track straightness. The introduction of an index of "passive swimming" made it possible to reveal episodes of immobility in water maze behavior of AF64A-injected animals. Unlike J-861, 17 beta-Estradiol almost completely eliminated these episodes. The newly developed indices (especially straightness) seem to be very useful in differentiating learning ability of rats from a decrease in their mobility in the Morris water-maze test, in particular, in case of the estrogens under study. PMID- 12754849 TI - [Intracortical synchronization of the epileptiform field potentials at different stages of ultrastructural reorganizations in neuronally isolates island in rats]. AB - A temporal delay (parameter of synchronization) between the incidence of epileptiform discharges in cortical sited located at a distance of 4 mm from each other was studied in rat intact cortex and neuronally isolated cortical slab using the cross-correlation function. Experiments were carried out at different stages of axonal sprouting. By 30 days of isolation, a significant increase in the number of boutons in the V cortical layer coincided with a significant decrease in the delay, whereas a reduction of the number of boutons by 90 days corresponded to its increase. These findings convincingly testify that the newly formed boutons form a basis for increase in synchronization and thus affect the epileptogenesis. The results obtained in this work and literature data suggest that under pathological conditions large pyramids of the V layer form a neuronal network which provides exclusively cortical synchronization of epileptiform field potentials. PMID- 12754850 TI - [Spontaneous unit activity of locus coeruleus neurons after destruction of some nuclei of the medulla oblongata]. AB - Bilateral lesions of the nuclei prepositus hypoglossi produced a more than twofold decrease in the mean frequency discharges in the neurons of the nucleus coeruleus. The number of neurons with burst activity and the number of polymodal neurons substantially increased. Lesion of the nucleus tractus solitarius resulted in an increase in the number of neurons with regular activity and certain decrease in the mean discharge frequency of coeruleus neurons. The results confirm the suggestion about a substantial role of the nucleus prepositus hypoglossi in relaying afferent effects to the activity of locus coeruleus neurons. PMID- 12754851 TI - [Rat brain reactions to acute brain stem lesion on dynamics of parameters of cerebral electric activity]. AB - Despite an old history of a question, adaptive brain reactions that develop after an acute brainstem lesion have not been adequately investigated. With the aim to study the central mechanisms of compensation/decompensation and to specify the character of involvement of orbitofrontal cortex and hippocampus into these processes the spatiotemporal organization of brain electric activity was analyzed in 8 rats before and after electrolytic brainstem lesion at the level of the lateral vestibular nucleus Deiters (VND). The electric activity was recorded from symmetric frontal and somatosensory cortical areas, hippocampal areas CA1, and intact VND. Spectralcoherent analysis showed that adaptive reactions are most clearly manifested by changes in the spatiotemporal organization of the theata activity: 1) early brainstemhippocampal synchronization of the electric activity in the frequency range of 6-7 Hz with subsequent involvement of anterior cortical regions is characteristic of survived animals; 2) independent hippocampal cortical hemispheric system of excitation in the frequency of 4-5 Hz precedes the fatal outcome. On the day before the fatal outcome the interhemispheric coherence in the orbitofrontal cortex dropped, which suggests the involvement of these brain regions into the processes of visceral regulation. PMID- 12754852 TI - [Local disinhibition with bicuculline does not break the trained relationship between afferent input and efferent output in cat motor cortex]. AB - Neurons of the cat motor cortex related to the lifting-withdrawal phase of forepaw placing reaction are preferentially activated by tactile stimulation of the dorsal surface of the forepaw. The placing reaction was altered in such a way that the innate "dorsal placing" was subjected to extinction and was substituted for the newly acquired conditioned reaction in response to the ventral side stimulation. This alteration of placing reaction led to the inversion of the innate input-output relationship in the motor cortex. The neurons related to forepaw lifting-withdrawal began to be activated by tactile stimulation of the ventral rather than dorsal forepaw surface. Local cortical disinhibition by bicuculline application at the recording site qualitatively changed neither normal input-output relationships nor inverse relationships after placing reaction alteration. This suggests that alteration of the sensorimotor coordination in cat motor cortex is underlain by changes in excitatory rather than inhibitory connections. PMID- 12754853 TI - [Rat behavior in the "open field" next day after haloperidol injection: dependence on experimental conditions]. AB - Wistar rats were injected with haloperidol (3.5 mg/kg) that resulted in a high level of cataplexy. Next day after haloperidol injection rat behavior was studied in the open field. The animals were divided in two groups. The first group of animals was tested in the daylight without additional illumination of the open field chamber. The second group was tested in a darkened room with additional intense illumination of the open-field center with a 60W bulb. The testing time was 240 s. The high level of the open-field locomotor activity in the first group was attributed to anxiety. The low level of locomotor activity in the second group was qualified as depressive state. PMID- 12754854 TI - [An experimental unit for the recording of the escape reaction of a ground snail to tactile stimulation]. AB - An original working experimental unit for noninvasive objective recording of the magnitude of escape reaction of a ground snail evoked by tactile stimulation is described. A. snail creeps upwards over the cylinder rotating around its horizontal axis. A watching device ensures a constant snail position relative to a light source and a photoelement. A device for tactile stimulation which provides graduated energy of an impact is constructed on the basis of the magnetic circuit of a loudspeaker. In response to a tactile stimulus a snail pulls in its feelers, head, and foot, and the area of snail's shadow decreases. These changes are indicated by the photoelement. PMID- 12754855 TI - [The theory of functional systems and probabilistic prognosis of behavior]. AB - The article is dedicated to possible probability prediction of behavior in system organization of behavioral acts. System mechanisms of anticipation of required results of behavioral activity by living organisms adapted to stable and changing conditions of life are discussed. The author proposes that in all forms of behavior, an organism strictly predicts parameters for satisfaction of dominating needs of results that constitute the goal of a given form of behavior. In author's opinion, the probabilistic prognosis concerns only methods, acts and attendant emotional states, as well as possible ways of attaining the results (i.e., the means rather than action parameters). PMID- 12754856 TI - ROC your organization! You can instigate Radical Organizational Change to achieve safety excellence. PMID- 12754857 TI - The young and the reckless? Injuries and death often occur when a teen is performing a task prohibited by state or federal laws. PMID- 12754859 TI - Bioaerosol evaluation in indoor environments. PMID- 12754858 TI - Facing the terror of nuclear terrorism. AB - As America prepares for homeland security and the response to terrorism, more occupational safety professionals may find themselves called upon to deal with terror in their own neighborhoods. While thousands of safety professionals are well trained technically to deal with many types of terrorism, they may not be well prepared to deal with the greater challenge, namely the terror of terrorism. Dealing with terror requires hearing and responding to people's feelings before providing technical answers. For safety professionals to be most effective in dealing with terrorism, they can benefit from more training on how to deal with terror. PMID- 12754861 TI - Duties and responsibilities of permit-required confined space entry team members. PMID- 12754860 TI - Gas detection for alternate-fuel vehicle facilities. AB - Alternative fuel vehicles' safety is driven by local, state, and federal regulations in which fleet owners in key metropolitan [table: see text] areas convert much of their fleet to cleaner-burning fuels. Various alternative fuels are available to meet this requirement, each with its own advantages and requirements. This conversion to alternative fuels leads to special requirements for safety monitoring in the maintenance facilities and refueling stations. A comprehensive gas and flame monitoring system needs to meet the needs of both the user and the local fire marshal. PMID- 12754863 TI - Heat stress precautions. PMID- 12754862 TI - Outdoor hazards: they're out to get you! PMID- 12754864 TI - Facing up to the ARC challenge. PMID- 12754865 TI - Selecting between supplied air and powered air. PMID- 12754866 TI - Ready for action. PMID- 12754867 TI - OSHA & light curtains. PMID- 12754868 TI - Playing on OSHA's team. PMID- 12754869 TI - [Novel approach to parathyroid adenoma: minimally invasive, focused, scan guided parathyroidectomy--experience from the first 100 cases]. AB - BACKGROUND: Traditionally, the surgical approach to parathyroid adenoma included formal bilateral neck exploration, inspection and evaluation of all four glands. Recently, following progress in the precision of pre-operative localization by sonography and scintigraphy and the availability of a real time PTH assay, focused, minimally invasive approaches to the removal of a single adenoma were proposed. We review our experience in the first 100 cases. METHOD: After localization of the suspected adenoma by TC-99m-MIBI scintigraphy and neck sonography, a second scan was performed just before surgery and the presumed site was marked on the patients skin. Under general anesthesia, via a limited incision, the suspected adenoma was excised and examined by a frozen section. RESULTS: Between July 1999 and August 2001, 97 patients (64 females and 33 males, mean age; 56 +/- 14, range 19-88) underwent 100 focused, minimally invasive, MIBI guided parathyroidectomies (3 patients were operated on twice due to a residual second adenoma). Pre-operative blood levels of calcium and PTH were 11.5 +/- 0.8 mg/dl and 140 +/- 90 pg/ml, respectively. In 93 cases, an adenoma was identified and excised (mean weight, 600 mg, range, 100-4900). Mean operative time, including frozen section was 66 +/- 39 minutes. The patients were discharged on the same day or on POD 1 (mean calcium level 9 +/- 0.9 mg/dl) and had normal calcium levels at the follow-up tests. In 2 cases, the scan was falsely negative for adenoma (positive for other thyroid pathology), but the adenoma was successfully excised according to the sonographic localization (overall success rate in the primary procedure, 95%). In 3 cases, pathologically proven enlarged parathyroid was excised, as localized by the scan, but hypercalcemia relapsed. The patients were re-operated in a focused fashion and a residual second adenoma was found (N = 2), or underwent formal exploration for hyperplasia of the remaining 3 glands (N = 1). In 2 more cases, no parathyroid tissue was found in the specimen. However, consequently, calcium levels normalized after surgery (N = 1) and a successful focused re-operation was performed after relocalization (N = 1). There were no significant post-operative complications. CONCLUSIONS: Focused MIBI guided parathyroidectomy is safe and efficient in most patients. Failures, which may be the results of erroneous diagnosis (hyperplasia vs. adenoma, 1%), residual additional adenoma (2%) or a false positive scan due to pathology in the thyroid gland (2%), can be treated safely and effectively in a second focused procedure. PMID- 12754870 TI - [Does hypothyroidism contribute to the etiology of primary open angle glaucoma or is it just a coincidence?]. AB - In a prospective study 83 consecutive patients with progressive primary open angle glaucoma (research group) and 62 patients scheduled for cataract surgery (control group), were evaluated for the presence of hypothyroidism by history, blood levels of TSH and free T4 when necessary. In the research group, nine patients (10.8%) had hypothyroidism, 6 of them already known. In group 2 only one patient (1.6%) had known hypothyroidism, this difference was statistically significant (p < 0.005). The association between primary open glaucoma and hypothyroidism is discussed. PMID- 12754871 TI - [Community acquired urinary tract infection among hospitalized children in northern Israel: pathogens, susceptibility patterns and urinary tract anomalies]. AB - BACKGROUND: Urinary tract infection (UTI) is one of the most common diseases in children. Vesicoureteral reflux (VUR) has been demonstrated in a substantial number of young children with UTI. Empiric antibacterial therapy is recommended before results of the urine culture are available in order to shorten the duration of the disease and prevent renal complications. OBJECTIVES: The aims of this study were to assess the prevalence and susceptibility patterns of UTI pathogens, and urinary anomalies in children admitted with UTI. METHODS: The study population included 151 children younger than 14 years admitted with first UTI. Renal ultrasound was performed in all the patients and voiding cystourethrography (VCUG) in children younger than 5 years. Dimercaptosuccinic acid (DMSA) scan was performed in children with vesicoureteral reflux. The data included age, sex, symptoms and signs, urinalysis, the pathogen and its sensitivity and the results of the imaging studies. RESULTS: A total of 119 patients (79%) were females. Gram negative rods caused 98% of the infections, of which Escherichia coli (87%) was the most prevalent pathogen, followed by Klebsiella pneumoniae (4%), and Proteus mirabilis (4%). The sensitivities to antibacterial agents were: Amikacin 100%, ceftazidime 97%, gentamicin 96%, ceftriaxone 96%, cefuroxime 95%, amoxicillin-clavulanate 84%, trimethoprim sulfamethoxazole 63%, cephalexin 58%, and ampicillin 28%. Renal US showed minor abnormalities in 24/149 (16%) and VCUG demonstrated vesicoureteral reflux in 38/127 (30%) patients. DMSA revealed renal scars in 7/28 (25%) children with vesicoureteral reflux. CONCLUSIONS: Overall Gram negative rods cause 98% of the UTI in hospitalized children in our area. E. coli is the leading pathogen and aminoglycosides and second or third generation cephalosporins are the most suitable agents for empiric therapy in UTI. A high incidence of renal scars in young children with vesicoureteral reflux was found. PMID- 12754872 TI - [Cardiovascular risk factors assessment in Rambam Center Preventive Medicine]. AB - Atherosclerosis, expressing itself as cardiovascular disease, is the first cause of death in Israel, in people over 35 years of age. The main risk factors for atherosclerosis are generally known, and amendable, resulting in a decrease in morbidity and mortality rates. In our present article we investigate the prevalence of major atherosclerosis risk factors in the population being examined in the Rambam Center of Preventive Medicine. This is a young population, usually considered healthy. The prevalence of the major risk factors is high, and a large portion of the population is at high risk of developing cardiovascular disease. Identifying the people at high risk of developing cardiovascular disease and of instigating preventive measures is of utmost importance. PMID- 12754873 TI - [Combined tests to measure functional capability in elderly people]. AB - BACKGROUND: Several diagnostic tests and questionnaires on the health and functional status of elderly people are used in the clinical practice. The evaluation procedures are geared to determine function and health issues distinct from that of the quality of life. The use of broad assessment complex protocols may be unrealistic, false or impractical. OBJECTIVES: The aims of the study were: a) to explore the relationship between a subjects perception and two functional tests, and b) to measure if Tinetti balance test and Berg balance test provide similar results. METHODS: Four common evaluation procedures were used prior and following 12 weeks of balance and exercise training: two are considered as objective tests (Tinetti and Berg), and two are considered as subjective questionnaires (Short-Form Health Survey--SF-36 and self satisfaction questionnaires). The procedures were applied to 38 independent elderly residents (aged 79 +/- 3.4) who live in a nursing home. RESULTS: Intraclass correlation coefficients were calculated to measure the reliability for the self-assessment of subjects who reported as "no change" "improvement" or as "worse". Pearson correlation coefficient was used to investigate the correlation between the tests. The self-assessed report differed from the clinical outcome tests. In the self-assessment questionnaire two subjects rated "worse" whereas no deterioration was noticed in Tinetti and Berg tests. Moreover, after 12 weeks of balance training, using a self-rated questionnaire, 7 subjects reported "no change", and in the clinical tests 13 subjects showed "no change". CONCLUSIONS: The combined test of Tinetti and SF-36 can provide an improved picture of the functional capability and perception in elderly persons in nursing homes. PMID- 12754874 TI - [Presumed Candida tropicalis endophthalmitis following anterior resection]. AB - We present a case of a 75 years old patient who developed a presumed Candida Tropicalis endophthalmitis after an anterior resection for rectal carcinoma. The pathogen was identified in blood cultures but not in vitreal cultures. Systemic anti fungal treatment brought no ocular improvement. After having a vitrectomy and intravitreal antifungal agent injection gradual improvement up to full resolution took place. To the best of our knowledge, this is the first report of a post surgical candida tropicalis endophthalmitis in a non-neutropenic patient. We suggest special attention to ophthalmic complaints in patients with systemic candidemia and prompt treatment according to ocular findings. PMID- 12754875 TI - [Nosocomial infections in internal medicine departments]. AB - BACKGROUND: Nosocomial infections are a major cause of morbidity and mortality among hospitalized patients. The use of new generation antibiotics of wide spectrum caused a higher rate of virulent infections. AIMS: a) To study the prevalence, pattern and etiology of nosocomial infections in patients hospitalized in departments of internal medicine. b) to determine the characteristics of high-risk patients, deserving special precautions. METHODS: All medical files of patients who were admitted to internal medicine departments during the years 1994-8 were reviewed. All patients with nosocomial infections were included according to specific criteria. RESULTS: Of 2789 medical files, 72 (2.6%) patients, randomly elected had 76 episodes of nosocomial infection. Urinary tract infection (40.8%), pneumonia (32.9%) and sepsis (9.2%) were the most frequent infections. Mortality was significantly higher in patients with nosocomial infections--48.6% compared to 5% in a control group (p < 0.001). E. Coli, Pseudomonas auroginosa, and Staphylococcus Aureus were the most frequent infecting bacteria. Empiric antibiotic therapy was used in 84.7% of the patients. Any bacterial isolation was found in 61.9% of the patients samples (including blood cultures, urine cultures etc). Old age, female gender, prolonged hospitalization, mechanical ventilation and indwelling urinary catheter were found as risk factors for nosocomial infections. CONCLUSIONS: 1. Nosocomial infections in internal medicine departments in Brazilai hospital were found to be less common than previously reported. 2. Five risk factors for nosocomial infections were correlated with a higher prevalence of the disease. 3. Mortality due to nosocomial infections is very common, probably due to inappropriate empiric treatment and high rate of bacterial resistance. 4. Simple preventive measures as well as immediate treatment of nosocomial infection with proper antibiotics are expected to decrease mortality. PMID- 12754876 TI - [Urinary tract infections in children--present and future]. AB - UTI in children can be the first sign of congenital renal malformations, especially reflux. Therefore, it is important to increase the awareness of the pediatrician in order to assist in the diagnosis and treatment, and in the evaluation of children with UTI. Sakran & al presented 151 children hospitalized in the Haemek Medical Center with UTI. Clinical characteristics of the children are described, distribution of uropathogens, their susceptibility to antimicrobials and the empiric antimicrobial therapy is assessed. In addition, the paper discussed the different image study procedures indicated in the evaluation of these children. PMID- 12754877 TI - [Oral cancer--molecular aberrations and tumor markers cellular aberrations]. AB - Squamous cell carcinoma (SCC) of the oral cavity is the sixth most frequent cancer in the world and approximately 30,000 new cases diagnosed annually in the United States, 60% occur in males and over 90% of the cases occur in patients over 45 years of age. The overall 5-year survival rate for patients with oral SCC is among the lowest for major cancers and has not changed during the past two decades. Assessing the severity of oral cancer prior to choosing the therapeutic strategy administered is of paramount importance. Molecular probing significantly increases the sensitivity and specificity of conventional histopathology. One example is the suitability demonstrated of TPS and Cyfra 21-1 in early detection of oral SCC and for its treatment monitoring. Another example is that inhibitors of SCC lesions attachment to laminin can now be tested as inhibitors of metastasis in experimental animal models. This type of information may eventually lead to the development of a new class of therapeutic agents that can alter the biologic behavior of oral squamous cell carcinoma in patients. In any case, oral cancer remains a disfiguring disease associated with a high mortality rate. PMID- 12754878 TI - [Experimental therapeutic modalities for retinitis pigmentosa]. AB - Retinitis Pigmentosa (RP) is a heterogeneous group of inherited diseases that cause retinal degeneration, often leading to blindness. Over the last decade, the advent of molecular biological techniques has greatly improved our understanding of the genetic and molecular processes underlying these diseases. This has prompted research efforts aimed at the development of novel therapeutic modalities for RP, and the purpose of this review is to present current trends in this field. In this first of two parts, advances in gene therapy, the use of neurotrophic growth factors, and attempts to alter disease progression by vitamins and nutritional modification are discussed. Experimental treatments involving surgical intervention will be addressed in a future, second part of this review that will include the topics of retinal and retinal pigment epithelium transplantation, the potential use of stem cells to replace degenerating retinal cells, and the development of "artificial vision" using electro-optical devices. The encouraging progress made in these various experimental directions lends hope that efficient treatment for at least some of the patients suffering from these blinding diseases is within sight. PMID- 12754879 TI - [Screening mammography for early diagnosis of breast cancer: facts, controversies, and the implementation in Israel]. AB - BACKGROUND: Breast cancer is the leading cancer among Israeli women. Mammography is the most widely used tool for early diagnosis of breast cancer. Eight published randomized controlled trials followed nearly 500,000 women over 7 to 18 years. Most trials found that screening mammography decreases breast cancer mortality by 20 to 40%. A recent study examined the methodology of the randomized clinical trials and found that most trials were flawed in the methodology of data collection and analysis in a way that might have influenced the results of those trials. PURPOSE: To review the studies, clarify the issues, and reach a conclusion regarding the utility of screening mammography in reducing breast cancer-related mortality in Israel. METHODS: A review of the world literature, and analyses of the Israeli data. RESULTS: Seven out of eight published randomized controlled trials found a significant decrease in breast cancer mortality among women who underwent screening mammography. A meta-analysis of the trials also supports the utility of screening mammography in decreasing breast cancer mortality. The criticism over the methodology of these trials does not necessarily invalidate their conclusions. CONCLUSIONS: The data indicate that screening mammography does indeed assist in early diagnosis, and most published studies show a significant reduction in breast cancer-related mortality in the screened population. Due to the high incidence of breast cancer in the Israel, especially among young women, the national screening program should continue. Moreover, consideration should be given to expanding it to women starting at age 45, instead of 50, as is practiced today. PMID- 12754880 TI - [Prenatal diagnosis of developmental congenital malformations--the limitations of ultrasound scanning]. AB - Prenatal diagnosis of congenital malformations is a major goal of obstetric sonography. Although significant progress has been made in the ability to detect fetal anomalies by ultrasound, some fetal anomalies cannot be detected during second trimester routine ultrasound scanning. Among the "undiagnosed anomalies" are many fetal anomalies that follow a developmental course in-utero and have a late-onset sonographic appearance, and hence cannot be diagnosed early in pregnancy or during the traditional mid-second trimester scan. Several mechanisms cause in-utero development of fetal malformations, and the developmental course of each fetal anomaly depends on the cause, mechanism, extent and timing of the insult. In some cases the destructive or disruptive event might occur at a relatively advanced gestational age and thus go undiagnosed. Some malformations are the result of an early insult but are manifested and detected late, while others have a "late onset". This concept of the developmental natural course of fetal anomalies in-utero, must be recognized and lead to a new nomenclature for fetal malformations. In this review we describe some developmental fetal malformations and discuss the clinical, diagnostic and medicolegal implications. PMID- 12754881 TI - [Method not stirrups--treating DDH according to Pavlik]. AB - Developmental dysplasia of the hip (DDH), previously termed "congenital dislocation of the hip" (CDH), encompasses a group of related pediatric disorders, some diagnosed soon after birth or later. These include clinical instability of the hip, with or without anatomical dysplasia, subluxation or dislocation. These days, such disorders are usually diagnosed soon after birth, and can develop to normal or worsen to severe disability in time. Most of the hips are not dislocated but all pathological hips show some degree of dysplasia and a minority is dislocated. The keystone to successful treatment of DDH is early diagnosis. Screening is successful only if followed by correct early treatment. Various methods and devices have been proposed for the treatment of DDH. After closed reduction of the hip is done, sometimes with violence, rigid fixation might cause avascular necrosis of the femoral head (AVN). These methods are used less and less, having been replaced by Pavliks method, whereby, if strictly used, the AVN rates are almost zero. PMID- 12754882 TI - [Acupuncture for nausea: how does it work?]. AB - Nausea, the unpleasant feeling that one is going to vomit, results from the stimulation of the Chemoreceptor Trigger Zone (CTZ) and Vomiting Center in the brain. Conventional medical therapy is not always effective, and medications often have severe adverse effects. Acupuncture is a treatment in which thin needles (diameter of 0.20-0.30 mm) are inserted into various points along the skin, according to energy channels (meridians) established thousands of years ago. The anti-emetic effects of acupuncture apparently stem from the resultant increase in hypophyseal secretion of beta-endorphins and ACTH, with subsequent inhibition of the CTZ and vomiting center. Acupuncture also affects the upper GI tract, decreasing acid secretion and repressing gastric arrhythmias. Clinical research has found this treatment modality to be effective for nausea, whether it be due to morning sickness in pregnant women, motion sickness in travellers, postoperative nausea or chemotherapy-induced nausea in cancer patients. PMID- 12754883 TI - [Ammonia and ammonium salts: remedy and poison, myth and time honored reality]. AB - The public interest in ammonia and its salts has risen due to the recent water crisis in Israel. The focus on their regulatory and environmental aspects has been intensified due to the elevated levels of ammonium salts in the national water system, resulting in a banning of water use in the Dan district. To the public it is commonly known that ammonia is toxic. Nevertheless, the medical view regarding ammonium salts is very different from that of ammonia gas. In contrast to the hazardous ammonia gas, its salts (such as ammonium bicarbonate) are considered to be much less toxic and are widely used as medicaments and food additives. Thus, although the presence of ammonium salts in the drinking water may indicate contamination, this is not a case of poisoning associated with toxic side effects. PMID- 12754884 TI - [Risk versus confidentiality: the therapist's responsibility for his psychiatric patient's acts]. AB - In this article we deal with the responsibility and liability of the therapist for aggressive and criminal acts perpetuated by his psychiatric patient against third parties. This issue has been the object of controversial discussion among professionals in the legal and medical field since the famous case of Tarasoff (this psychiatric patient killed his former girlfriend after he informed his therapist of his intention to harm her. The therapist was sued because he did not warn nor protect the potential victim and did not prevent the criminal act). We describe the situation in the U.S. before Tarasoff and the implications of the decision of the Supreme Court of California in 1974-76 which dealt with this case and broadened the therapist's responsibility towards third parties. We also analyze the actual legal situation in Israel on this issue, its disadvantages and shortcomings. We suggest some proposals for improvement, together with some operational measures to cope with this complex issue including a thorough evaluation of the patient's clinical state, identification of the potential victim, warning and involvement of the relevant authorities, and hospitalization if needed. PMID- 12754885 TI - [Standpoint on the use of angiotensin receptor blockers in the treatment of hypertension]. AB - Angiotensin receptor blockers (ARBs) have gained widespread use in clinical medicine during the past decade. Several large, prospective and randomized multi center trials have led us to reconsider the role of ARBs in the treatment of hypertension. Firstly, in view of the favorable safety and side effect profile of ARBs, we recommend their use in hypertensive subjects in whom ACE inhibitors are indicated but are unable to tolerate agents of this type due to intractable cough. Secondly, in light of the results of the RENAAL and IDNT studies, we consider ARBs as the drug of choice in diabetic subjects with hypertension and proteinuria (> 300 mg/L). Thirdly, we view ACE inhibitors and ARBs as equally adequate for the treatment of diabetic patients with hypertension and microalbuminuria and recommend the use of the maximal allowable doses of these drugs in such patients. Finally, older hypertensive individuals with left ventricular hypertrophy should receive either ACE inhibitors or ARBs, as these drug classes presently appear to provide better overall protection than beta blockers or calcium channel blockers in this particular subgroup of patients. PMID- 12754886 TI - [Impressions and lessons from the National Forum on quality control improvement in healthcare]. PMID- 12754887 TI - [Current problems of the public health X-ray service of the Russian Federation and ways of their solution (according to the analysis materials of their activities of the service in 2001 and its tasks in 2002]. PMID- 12754888 TI - [Resolution of the first meeting of chief roentgenologist (radiation diagnosticians) of the subjects of the Russian Federation, April 16, 2002]. PMID- 12754889 TI - [Brain CT diagnosis in infants of the first year of life who have congenital heart diseases]. AB - Computed tomography (CT) was used to study the brain in 73 infants of the first year of life who had different congenital heart diseases (CHD). CT was performed on a HiSpeed CT/i spiral computer tomograph (the firm "CE") and a C-150 XP electronic radiation tomograph (the firm "Imatron"). The capacities of the technique in the diagnosis of brain lesion were explored in CHD infants of the first year of life. The studies indicated that the commonest abnormalities in CHD infants of this age were hydrocephalus frequently concurrent with congenital malformations, as well as diffuse and focal changes in the brain. PMID- 12754890 TI - [Digital X-ray detection of extrapulmonary pathology during chest screening]. AB - The outcomes of digital roentgenography are analysed at screening examinations of members of chest 8000 men: with a pathology lung--83, cardiovascular system- 1625, thoracal spine--2034. The outcomes bear to high quality of the image not only lung, but also hearts, aorta, thoracal spin, allow to prefer digital roentgenography of members of a chest as to effective procedure screening of surveys of the population. PMID- 12754891 TI - [Use of magnetic resonance imaging in surgical hepatology]. AB - Three hundred and ninety eight patients with different diseases of the liver and biliary tract were examined on the basis of the complex use of latest technologies of magnetic resonance imaging (dynamic contrast MRI, MR-arterio- and venoportography, MR-cholangiopancreatography), which yield all necessary information during one study to design surgical treatment. The study was conducted by using a Magnetom Vision 1.5T apparatus (Siemens). The advantages of the technique over invasive X-ray studies are shown. Different MRI procedures have been ascertained to be highly diagnostically effective in evaluating hepatic and biliary diseases. PMID- 12754892 TI - [Magnetic resonance imaging of the ankle joint and foot: possible diagnostic errors]. AB - The paper deals with the visualizing study of the ankle joint and foot by MRI and with the problems in the interpretation of magnetic resonance images in health. For this, 50 healthy volunteers without diseases and lesions of the ankle joint and foot were examined. The study was performed by using flexible superficial coils and T1-, T2-, and proton-weighed pulse-sequences in the orthogonal projections. The articular surfaces and cavity of the ankle joint were evaluated. The specific features of visualization of the muscles and tendons of this area and the pattern of fluid under their membranes were explored. The typical location of the "magic corner" phenomenon was revealed. The individual specific features of identification of the ligaments of the ankle joint and foot and plantar aponeurosis were defined. The features of visualization of bones simulating abnormalities were studied. A category of normalcy in the MRI of the ankle joint and foot was formulated. PMID- 12754893 TI - [The capacities of autoleukocyte-labelled scintigraphy in the detection of foci of inflammation and suppuration]. AB - The diagnostics of occult inflammation foci up to day remains the major and complicated problem in almost each clinical discipline. Whole-body scintigraphy and SPECT studies with 99mTc-autoleucocytes show silent foci of inflammation and infection. 68 patients (24 females and 44 males) aged 14-59 years with non localized inflammatory diseases were evaluated. Each patient underwent routine clinical and laboratory investigation. But because of the inadequacy of other diagnostic procedures, the Tc-99m-WBC scans may be of great clinical importance. For the determining of the SPECT of thorax were performed using Tc-99m-HMPAO (hexametylpropilenaminoxime) labeled autoleucocytes. In 1, 3 and 24 hours post injection inflammatory foci were seen as the areas of pathogenic activity. In 24 patients the circumscribed activity seen in relation to heart convincingly demonstrated endo- and/or myocarditis. 16 patients showed acute inflammation in postoperative wound, empyema of pleura and mediastinitis as the consequences of cardiosurgery. In 16 patients whole-body scanning and SPECT of the head revealed chronic infection in nasopharynx and accessory nasal sinuses dental infection and the rest 7 patients had abdominal and pelvic inflammatory diseases, such as ulcerative colitis, intraabdominal and retroperitoneal septic foci, tubo-ovarial inflammatory disease. The data demonstrated that Tc-99m-leucocytes imaging is a highly sensitive and specific for the detection of inflammatory diseases of different localization. PMID- 12754894 TI - [Use of computed tomographic image reconstruction in some abnormal maxillofacial changes]. PMID- 12754895 TI - [Potentialities of layer X-ray study in the solution of tactic issues of perosseous synthesis]. PMID- 12754896 TI - [The specific features of magnetic resonance imaging of uterine myomas with cystic degeneration]. PMID- 12754898 TI - Developments in medical education. PMID- 12754897 TI - [Experience in organizing a complex radiation diagnosis of tumors at therapeutical institutions of a large industrial region]. PMID- 12754899 TI - Task force report. Part II: Barriers to professionalism. PMID- 12754900 TI - It's more than a computer can deliver: gross anatomy--a rite of passage and a right to learning. PMID- 12754901 TI - A celebration of education--Wisconsin physicians and their Wisconsin Medical Society. PMID- 12754902 TI - The Medical College of Wisconsin's program to strengthen geriatrics education. AB - Medical care for geriatric patients requires physician training that promotes the acquisition of attitudes, knowledge and skills that will permit future practitioners to meet the health needs of increasing numbers of aged patients. MCW has strengthened its traditional curriculum by focusing on student attitudes in the early pre-clinical years through outreach and interest groups programs. Knowledge is integrated throughout the 4-year curriculum using our aging virtual patients. These patients are a teaching resource to the entire faculty. Attitudes, knowledge, and skills in geriatrics are further developed through an M3 geriatrics medicine option and the M4 Integrated Selective. Geriatric-specific skills are emphasized through the use of standardized patients and objective structured clinical examinations in the M4 Selective. It is anticipated that these students efforts will create interest in a novel residency experience (Med Ger) that will ensure that upon successful completion of the program, residents are expert in geriatric medicine practice and meet criteria for board certification in geriatric medicine. PMID- 12754903 TI - A view from the clinical campus: a partnership of an academic medical center with an integrated delivery system. PMID- 12754904 TI - A faculty leadership development program at the Medical College of Wisconsin. AB - A faculty leadership development program has been initiated at the Medical College of Wisconsin in collaboration with faculty from the University of Wisconsin, Milwaukee, School of Business Administration. The program's goal is to impart business-related knowledge and develop leadership skills among the senior and selected junior faculty members. The course is given over nine days in segments of three days over a five-month period. So far it has been given three times. Course evaluations by attendees indicate that the course is highly regarded and they consider it very useful in developing their managerial and leadership skills. This article describes how a free-standing medical school can collaborate with a business school to develop and offer a program customized to meet the management and leadership training needs of its faculty. PMID- 12754905 TI - Matriculating student perceptions of changes to the admissions interview process at the University of Wisconsin Medical School: a prospective, controlled comparison. AB - PURPOSE: The purpose of this study was to assess whether changes in the admissions interview process improved matriculating students' perceptions of the quality of the admissions interview process. METHODS: We surveyed matriculating medical students for a 3-year period. Over this period, the admissions process and procedures went through a review and then a re-engineering. The survey provided 1 year of baseline data (while the review was undertaken), 1 year of data as recommendations from the review were progressively implemented, and 1 year of data on full implementation of recommendations. RESULTS: From baseline to full implementation, there were statistically significant increases (p < .05) in the percentage of matriculating students who found the University of Wisconsin (UW) Medical School interview process useful (31% increase), thorough (50% increase) and better than that of other medical schools (28% increase). There was also a statistically significant decrease in the percentage of matriculating students who found the UW Medical School interview process to be less impressive than other schools (29% decrease) and in need of improvement (45% decrease). EDUCATIONAL SIGNIFICANCE/CONCLUSIONS: Changes made in the UW Medical School's interview process yielded significant increases in perceptions of the quality of the experience by matriculating students. Since interviewing is expensive for both the institution and the applicant, it should have a clear purpose. The manner in which interviews are conducted should be critically reviewed periodically to ensure that the interview continues to meet its intended needs. PMID- 12754906 TI - A new program to reward and retain volunteer clinical faculty at the Medical College of Wisconsin. AB - BACKGROUND AND PROBLEM: Volunteer clinical faculty (VCF) are vital to the educational mission of medical schools. At the Medical College of Wisconsin (MCW), VCF are increasingly relied upon to meet clinical training needs in medical student and resident education. However, many VCF receive little or no preparation to excel in their teaching roles, and they are under increasing time demands that limit their availability to teach. METHODS: Beginning in 2001, the primary care departments at MCW began a series of initiatives called "ExCEED" to promote VCF teaching excellence and efficiency through two main program components: Advisory Councils, made up of VCF leaders, and Support Services, such as web-based resources and teaching workshops. RESULTS: Preliminary ExCEED findings show that VCF have acquired important knowledge, skills, and tools that have better prepared them for their teaching roles. CONCLUSION: ExCEED is a systematic, multi-method approach to engage VCF that is positively influencing the clinical education of MCW students and residents. PMID- 12754907 TI - Graduate medical education at the Medical College of Wisconsin: new initiatives to respond to the changing residency training environment. AB - Nationally, Graduate Medical Education (GME) is facing a series of challenges. These include cutbacks in Medicare funding, major changes in accreditation standards requiring education in and assessment of "general (core) competencies," and reduction in housestaff work hours. GME at the Medical College of Wisconsin (MCW) is managed by a consortium called the Medical College of Wisconsin Affiliated Hospitals, Inc. (MCWAH), which is comprised of 13 health care institutions in Southeastern Wisconsin. The general competencies required by the Accreditation Council for Graduate Medical Education (ACGME) include six focal areas: (1) patient care; (2) medical knowledge; (3) professionalism; (4) interpersonal and communication skills; (5) practice-based learning and improvement; and (6) systems-based practice. Traditionally, the GME programs have focused on training and assessment specific to patient care and medical knowledge, but have limited emphasis on the other four. To address this gap, MCWAH has launched several initiatives to enhance teaching and assessment of the other four competencies. An on-line residency management system marketed by New Innovations of Toledo, Ohio is being used to provide a web-based residency management system, allowing the faculty and residents to evaluate one another at the end of each rotation. Faculty development programs for residency program directors have been initiated to ensure they have the knowledge and skills associated with teaching and assessing the core competencies. We are now piloting a 360-degree evaluation system to include evaluations of residents and faculty by co-workers and patients. The ACGME is in the process of mandating reduced duty hours for the housestaff. As a result, residents will have less time for direct patient care responsibilities with more intensive use of other education and training strategies to ensure that they become independent specialists. GME is undergoing a major paradigm change, and MCWAH remains on the cutting edge in responding to the challenges. PMID- 12754908 TI - The role and future of standardized patients in the MCW curriculum. PMID- 12754909 TI - Personal digital assistants herald new approaches to teaching and evaluation in medical education. AB - Since its arrival in 1994, the personal digital assistant (PDA) has made significant inroads in the handheld industry, with 50% of physicians anticipated as users by 2005 due to its functionality as a point-of-care medical informatics tool. However, its use in medical education is less well documented. Since 1998, PDAs have been used at Medical College of Wisconsin (MCW) as both a teaching and an evaluation tool for medical student and resident education. This article highlights the use of the PDA in medical education and describes current applications for monitoring clinical experiences of students/residents, and teaching resources for hypertension, cardiac auscultation, and community health. MCW's experiences with the PDA as a real time teaching and data collection tool serves as a model for other medical schools and for our students who are educated in the importance of self-monitoring one's clinical experiences and the need for continuous improvement as future physicians. PMID- 12754910 TI - Are Wisconsin physicians knowledgeable about male osteoporosis? AB - It is unknown if recent research defining the relatively frequent occurrence and pathophysiology of male osteoporosis has been conveyed to clinicians. To assess if physicians have incorporated such information, 5646 licensed Wisconsin physicians received a 1-page survey consisting of 14 statements regarding general knowledge, diagnosis, and treatment of osteoporosis in men. Twenty-six percent (1488) responded; 69% (1033) were family physicians or internists, 7% (106) orthopedic surgeons, and 5% (63) endocrinologists or rheumatologists. Of these physicians, 61% to 78% recognized that osteoporosis is not rare in men. Most (68% 97%) agreed that low-trauma fracture or corticosteroid therapy initiation are indications for bone mass measurement. Treating osteoporosis at a T-score of -2.5 was accepted by 79% to 89% of responders. Overall these physicians: (1) recognize osteoporosis as an important disease in men; (2) accept corticosteroid therapy and prior low-trauma fracture as indications for bone mass measurement; and (3) utilize T-scores in treatment decisions. These data suggest that knowledge regarding male osteoporosis has been conveyed to practicing physicians. Evaluation of methods by which physicians will translate this knowledge into action, thereby optimizing patient care, are needed. PMID- 12754911 TI - Future physicians need hands-on education. PMID- 12754913 TI - 2003 revised Wisconsin Medical Society constitution and bylaws. PMID- 12754912 TI - MetaStar's quality initiatives for 2003-2006. PMID- 12754914 TI - [Political, social and ethical perspectives of research in a era of barbarism]. PMID- 12754915 TI - [Occupational accidents with sharp instruments in nursing workers]. AB - The present study analyzed the accidents happened at work with the nursing staff of a public hospital, provoked by sharp instruments. All the nursing staff composed the study population. In the collection of data two instruments were used: a questionnaire designed to collect data referring to all working accidents that happened in the year of 1996; the second was an interview accomplished with these workers that suffered accidents with sharp instruments. The results showed that the nursing staff health is under risk with sharp instruments is involved in the accidents. For these accidents, the greater risk coefficients were for nursing auxiliary. High subnotification was verified for those accidents. The relative results of the interaction of the nursing staff working with sharp instruments indicate the great exposure to the biological risks and the serious diseases. PMID- 12754916 TI - [Development of an instrument for evaluating the moving and transfer of patients: an ergonomic approach]. AB - Nursing personnel are at a high risk from work-related musculoskeletal disorders, especially back symptoms. Handling patients has been attributed as one of the factors playing an important role in the aetiology of occupational low back pain. The aim of the study was to construct an instrument to evaluate the ergonomic risks during patient-handling tasks. The instrument was developed with an ergonomic approach involving the equipment, the tasks, the environment and the personnel. PMID- 12754917 TI - [Nursing and costs management]. AB - This work intend to broach structural elements that motivate institutions and health professionals to know and develop studies about hospital costs, emphasize the subject importance for the nursing professional. PMID- 12754918 TI - [Perroca's instrument for patient classification: reliability test for evaluators agreement--correlation]. AB - The purpose of this research was to check the reliability of Perroca's instrument model for classifying patients, based on the particular needs of nursing care. This patients' classification instrument was applied in 50 patients randomly chosen by the nurses of the General Intensive Care Unit of a medical school hospital. Pearson's quotient correlation was used to analyze the concordance degree of the total scores by the nurses. The reliability analysis showed a high and positive correlation in relation to scores given by the nurses. PMID- 12754919 TI - [Utilization of paraformaldehyde tablets by Brazilian health institutions-II]. AB - The descriptions of the part I of this study showed that Paraformaldehyde tablets in environmental conditions was used in a brazilian reality as the inadequate choosing way for most of the researched Institutions. This articles shows how this chemical agent is used outstanding problems as the lack of a theoretical base and process validation compromiting, this way, the safety of the sterilization. The "sterilization" conditions of most of the Institutions didn't include temperature, increment of the relative humidity and didn't make the adapted quantification of the tablets. It was observed that the researched Institutions don't have based parameters that guide the reutilization of a same group of Paraformaldehyde tablets in the sterilization processes. The use of Individual Protection Equipment is quite valued in handle of this chemical agent. PMID- 12754920 TI - [Maternal care of low-birth weight infants]. AB - This is an exploratory and descriptive study which aimed at characterizing mothers' accounts on the care given to the health of children with low weight at birth during their first and second years of life by means of home interviews with a group of eleven mothers of children from Ribeirao Preto--SP. The mothers pointed to difficulties with breastfeeding, the babies' diet and medication use. They also referred to concerns about intercurrences, re-hospitalizations and psychomotor development, which results in the need for professional support in order to evaluate and follow the growth and development process of children within families' everyday life. PMID- 12754922 TI - [Strategies developed by the elderly for controlling arterial hypertension]. AB - We objectified to investigate facing strategies elaborated by old-aged people which were attacked by arterial hypertension, being participants of a self helping group, in a therapeutic accompaniment. The facing mechanisms corresponded to faith in God, family support, occupational activities accomplishment, leisure activities and group participation. Some old-aged people mentioned the family structure as the sustentation pillar for a better therapeutic following, making the elaboration of adaptative answers possible. We concluded that, in spite of all the felt and referred losses, the old-aged people searched to active mechanisms that propitiate adaptative answers to the health-disease situation. PMID- 12754921 TI - [Factors that support and do not support the staying of mothers in a hospital pediatric unit]. AB - The factors that do or don't support the permanence of accompanying mothers in a pediatric unit were evaluated. Six interviews were held with the mothers and analyzed according to the themes. The factors that support the accompanying mother are: count on presence of other family members (mainly the child's father), the possibility of free meals at the hospital; trust the hospital team and being heeded by this team. The factors that don't support the mother are: child's illness; worry about other responsibilities (other children and house chores) impossibility to sleep and rest; lack of attention from the hospital team and the impossibility to receive visits. PMID- 12754923 TI - [Every day use of phytotherapy in a rural population]. AB - The aims of this study are to analyze the use of phytotherapy and the context of its utilization in a rural area population--MG; to identify the herbs used with more frequency and their forms of use; to compare the results of medicinal herbs with that described in literature. Thirty three people were interviewed in 1997. About the phytotherapy herbs known by them 106 different species were quoted, and 60 of them were effectively used in their routine. the transmission of knowledge related to the use of the herbs occurred basically from grandparents to the children. The phytotherapy was used by the population studied with similar purposes described in the literature aiming the prevention and treatment of diseases. PMID- 12754924 TI - [Preventive screening for cancer of the uterine cervix: awareness and meaning for employees in a public nursing school]. AB - This is an exploratory descriptive study, carried out at a nursing school, in which 63 non-teacher workers participated. Its objectives were to check their knowledge about screening of cervix-uterine cancer and to know their feelings and expectations as they undergo this test. A questionnaire was used to collect data and the last question was recorded and analyzed using the social phenomenology theoretical framework. The answers provided information which made it possible for us to rethink about adequate assistance guided by humanized actions considering the being in this existential entirety and belonging to a social economical cultural context. PMID- 12754925 TI - [Modern HPLC in drug analysis]. AB - High performance liquid chromatography found wide-spread utilization almost in all fields of pharmaceutical analysis since the 1980s. The article summarizes the new knowledge and interesting findings about HPLC used in the analysis of drugs during the last three years with a supplement of particular applications in pharmaceutical analysis. PMID- 12754926 TI - [The Beilstein CrossFire Information System and its use in pharmaceutical chemistry]. AB - The Beilstein CrossFire database system is one of the most popular, most extensive, and most accomplished systems presenting information about chemical substances, including drugs, and covering the basic identification, physico chemical, ecological, and pharmacological data. The present article offers an overview of historical evaluation of the Beilstein CrossFire database system, beginning from its founder Prof. Friedrich Conrad Beilstein till the most recent prospects about the connection of classical data with full-text databases. The article also offers a review of the development of electronic information systems in the Slovak Republic, a definition of basic functionality of the Beilstein CrossFire database system, and an example of targeted selection of active substances structurally similar to novel COX-2 inhibitors having an approved anti inflammatory activity. PMID- 12754927 TI - [Advances in the development of antitubercular agents containing no heterocyclics in the pharmacophore. Part 1. Derivatives of carboxylic acid]. AB - The present paper is the twentieth communication in a series of review papers on the development of antituberculotics and the ninth communication summarizing the work in the development of antituberculotics within recent 15 years. The first seven communications were devoted to heterocyclic compounds, the eighth to substances containing an alkylsulfanyl group in the pharmacophore. The present paper summarizes information about cyrboxylic acids, their amides, anilides, hydrazides, thioamides, thioanilides, thiohydrazides, and amidines. The communication includes papers published after 1980 in order to link up with a review paper devoted to the same topic published in the present series in 1982. PMID- 12754928 TI - [Pharmaceutical significance of Allium sativum L. 4. Antifungal effects]. AB - The recent decades saw an increase in the number of systemic fungal diseases and improvements in the identification of their causative agents. There has been an intensive search for new drugs which would be more effective and less toxic than those already in use. From this aspect, attention has been paid also to garlic- its extracts and individual components, i.e., allicin, ajoen, polysulfides, essential oil. New experimental knowledge confirms a significant antifungal activity of sulfurous compounds of garlic. The paper also mentions a possible use of employing garlic extracts or essential oils in food industry as an alternative way of protection of foodstuffs from contamination with fungi. PMID- 12754929 TI - [Penetrometry in evaluation of flow and structure of powdered substances]. AB - Usability of modified penetrometry is examined and significant relationships with routinely used characteristics of flowability and structure of powdered substances are searched for in size fractions of sodium chloride, potassium chloride, sodium citrate, and potassium citrate. A systematic study of interrelationships between properties made it possible to achieve sufficiently reproducible results without a necessity of consolidation. By means of factor analysis, a connection between penetrometry, bulk flow rate, and Kawakita constant was demonstrated. For multidimensional linear regression for estimation of penetrometry, the significance of examined properties decreases in the order Kawakita-constant > loose bulk density > total porosity > bulk flow rate > mass flow rate. The most important item of knowledge gained in the study is the dependence of penetrometry on a combination of loose bulk density and Kawakita constant. PMID- 12754930 TI - [Rheologic properties of Vivastar P 5000 hydrogel]. AB - Within the framework of pre-formulation studies, the paper evaluates the rheological properties of hydrogels whose gel-forming substance is a polymer supplied by the firm J. Rettenmaier & SohneGMBH + CO. It is VIVASTAR P 5000, a modified derivate of carboxymethyl starch. The evaluation of rheological properties included hydrogels with 1.5, 2.0 and 2.5 w % of VIVASTAR P 5000. The highest viscosity was shown by the hydrogel with 2.0% concentration of the polymer. The system without and with the model drug with a local anaesthetic effect was thixotropic. From the viewpoint of topical administration, VIVASTAR P 5000 in 2.0% is optimal for hydrogel formulation. PMID- 12754931 TI - [Effect of saccharides in the teicoplanin stationary phase on the separation of enantiomers of the phenylcarbamic type using HPLC]. AB - The paper presents the results obtained with the use of two chiral stationary phases (CSP) based on a macromolecular antibiotic agent--teikoplanin containing saccharide parts (CHIROBIOTIC T) and teikoplanin without saccharide parts (CHIROBIOTIC TAG). Racemic mixtures of 1-methyl-2-piperidinoethyl esters of 2-, 3 , and 4-alkoxyphenylcarbamic acids were examined. The investigation included interactions between separated substances and CSP, and the effect of separation of the enantiomers under study on the value of the differentiation factor (Rij) under identical chromatographic conditions with the use of the method of high performance liquid chromatography (HPLC). On the basis of the obtained results, it is possible to report that CSP-CHIROBIOTIC TAG is more advantageous for the substances of the type of 1-methyl-2-piperidinoethyl esters of 2-, 3-, and 4 alkoxyphenylcarbamic acids, because it does not contain a saccharide part, which decreases the possibility of non-polar interactions which exert a negative effect on the Rij value. PMID- 12754932 TI - What do we mean by biological complexity? AB - The purpose of the present paper is to offer a precise definition of the concepts of integration, emergence and complexity in biological networks through the use of the information theory. If two distinct properties of a network are expressed by two discrete variables, the classical subadditivity principle of Shannon's information theory applies when all the nodes of the network are associated with these properties. If not, the subadditivity principle may not apply. This situation is often to be encountered with enzyme and metabolic networks, for some nodes may well not be associated with these two properties. This is precisely what is occurring with an enzyme that binds randomly its two substrates. This situation implies that an enzyme, or a metabolic network, may display a joint entropy equal, smaller, or larger than the corresponding sum of individual entropies of component sub-systems. In the first case, the collective properties of the network can be reduced to the individual properties of its components. Moreover, the network is devoid of any information. In the second case, the system displays integration effects, behaves as a coherent whole, and has positive information. But if the joint entropy of the network is smaller than the sum of the individual entropies of its components, then the system has emergent collective properties and can be considered complex. Moreover, under these conditions, its information is negative. The extent of negative information is enhanced if the enzyme, or the metabolic network, is far away from equilibrium. PMID- 12754933 TI - Does the size of small objects influence chemical reactivity in living systems? AB - Previous theoretical works showed that chemical reactions in micro- and nano droplets, bubbles and solid particles were strongly affected by their confinement. In particular, the smallness of the systems leads to high internal pressure compared to the external pressure, which then significantly modifies the values of chemical equilibrium and kinetic constants. In addition, surface tension or surface stress, reactional dilatation and surface charge play also a major role on the chemical reactivity. As living systems are also made of very complex dispersed subsystems, i.e. organelles, it seemed obvious to illustrate our theory by some biological actual examples encountered in pulmonary alveolae, in vacuolae and in medical applications, such as dissolution of gallstones. PMID- 12754934 TI - Biological processes in organised media. AB - Embedding a simple Michaelis-Menten enzyme in a gel slice may allow the catalysis of not only scalar processes but also vectorial ones, including uphill transport of a substrate between two compartments, and may make it seem as if two enzymes or transporters are present or as if an allosterically controlled enzyme/transporter is operating. The values of kinetic parameters of an enzyme in a partially hydrophobic environment are usually different from those actually measured in a homogeneous aqueous solution. This implies that fitting kinetic data (expressed in reciprocal co-ordinates) from in vivo studies of enzymes or transporters to two straight lines or a sigmoidal curve does not prove the existence of two different membrane mechanisms or allosteric control. In the artificial transport systems described here, a functional asymmetry was sufficient to induce uphill transport, therefore, although the active transport systems characterised so far correspond to proteins asymmetrically anchored in a membrane, the past or present existence of structurally symmetrical systems of transport in vivo cannot be excluded. The fact that oscillations can be induced in studies of the maintenance of the electrical potential of frog skin by addition of lithium allowed evaluation of several parameters fundamental to the functioning of the system in vivo (e.g., relative volumes of internal compartments, characteristic times of ionic exchanges between compartments). Hence, under conditions that approach real biological complexity, increasing the complexity of the behaviour of the system may provide information that cannot be obtained by a conventional, reductionist approach. PMID- 12754935 TI - Actin-based motility as a self-organized system: mechanism and reconstitution in vitro. AB - Site-directed actin polymerisation in response to signalling is responsible for the formation of cell protrusions. These elementary 'actin-based motility processes' are involved in cell locomotion, cell metastasis, organ morphogenesis and microbial pathogenesis. We have reconstituted actin-based propulsive movement of particles of various sizes and geometries (rods, microspheres) in a minimum motility medium containing five pure proteins. The ATP-supported treadmilling of actin filaments, regulated by Actin Depolymerizing Factor (ADF/cofilin), profilin and capping proteins provides the thermodynamic basis for sustained actin-based movement. Local activation of Arp2/3 complex at the surface of the particle promotes autocatalytic barbed end branching of filaments, generating a polarized arborescent array. Barbed end growth of branched filaments against the surface generates a propulsive force and is eventually arrested by capping proteins. Understanding the mechanism of actin-based movement requires elucidation of the biochemical properties and mode of action of Arp2/3 complex in filament branching, in particular the role of ATP binding and hydrolysis in Arp2/3, and a physical analysis of the movement of functionalised particles. Because the functionalisation of the particle by an activator of Arp2/3 complex (N-WASP or the Listeria protein ActA) and the concentrations of effectors in the medium are controlled, the reconstituted motility assay allows an analysis of the mechanism of force production at the mesoscopic and molecular levels. PMID- 12754936 TI - Genetic regulation networks: circuits, regulons and attractors. AB - We deal in this paper with the concept of genetic regulation network. The genes expression observed through the bio-array imaging allows the geneticist to obtain the intergenic interaction matrix W of the network. The interaction graph G associated to W presents in general interesting features like connected components, gardens of Eden, positive and negative circuits (or loops), and minimal components having 1 positive and 1 negative loop called regulons. Depending on parameters values like the connectivity coefficient K(W) and the mean inhibition weight I(W), the genetic regulation network can present several dynamical behaviours (fixed configuration, limit cycle of configurations) called attractors, when the observation time increases. We give some examples of such genetic regulation networks and analyse their dynamical properties and their biological consequences. PMID- 12754937 TI - Stochastic models for circadian rhythms: effect of molecular noise on periodic and chaotic behaviour. AB - Circadian rhythms are endogenous oscillations that occur with a period close to 24 h in nearly all living organisms. These rhythms originate from the negative autoregulation of gene expression. Deterministic models based on such genetic regulatory processes account for the occurrence of circadian rhythms in constant environmental conditions (e.g., constant darkness), for entrainment of these rhythms by light-dark cycles, and for their phase-shifting by light pulses. When the numbers of protein and mRNA molecules involved in the oscillations are small, as may occur in cellular conditions, it becomes necessary to resort to stochastic simulations to assess the influence of molecular noise on circadian oscillations. We address the effect of molecular noise by considering the stochastic version of a deterministic model previously proposed for circadian oscillations of the PER and TIM proteins and their mRNAs in Drosophila. The model is based on repression of the per and tim genes by a complex between the PER and TIM proteins. Numerical simulations of the stochastic version of the model are performed by means of the Gillespie method. The predictions of the stochastic approach compare well with those of the deterministic model with respect both to sustained oscillations of the limit cycle type and to the influence of the proximity from a bifurcation point beyond which the system evolves to stable steady state. Stochastic simulations indicate that robust circadian oscillations can emerge at the cellular level even when the maximum numbers of mRNA and protein molecules involved in the oscillations are of the order of only a few tens or hundreds. The stochastic model also reproduces the evolution to a strange attractor in conditions where the deterministic PER-TIM model admits chaotic behaviour. The difference between periodic oscillations of the limit cycle type and aperiodic oscillations (i.e. chaos) persists in the presence of molecular noise, as shown by means of Poincare sections. The progressive obliteration of periodicity observed as the number of molecules decreases can thus be distinguished from the aperiodicity originating from chaotic dynamics. As long as the numbers of molecules involved in the oscillations remain sufficiently large (of the order of a few tens or hundreds, or more), stochastic models therefore provide good agreement with the predictions of the deterministic model for circadian rhythms. PMID- 12754938 TI - Emergence of complex behaviour from simple circuit structures. AB - The set of (feedback) circuits of a complex system is the machinery that allows the system to be aware of the levels of its crucial constituents. Circuits can be identified without ambiguity from the elements of the Jacobian matrix of the system. There are two types of circuits: positive if they comprise an even number of negative interactions, negative if this number is odd. The two types of circuits play deeply different roles: negative circuits are required for homeostasis, with or without oscillations, positive circuits are required for multistationarity, and hence, in biology, for differentiation and memory. In non linear systems, a circuit can positive or negative (an 'ambiguous circuit', depending on the location in phase space. Full circuits are those circuits (or unions of disjoint circuits) that imply all the variables of the system. There is a tight relation between circuits and steady states. Each full circuit, if isolated, generates steady state(s) whose nature (eigenvalues) is determined by the structure of the circuit. Multistationarity requires the presence of at least two full circuits of opposite Eisenfeld signs, or else, an ambiguous circuit. We show how a significant part of the dynamical behaviour of a system can be predicted by a mere examination of its Jacobian matrix. We also show how extremely complex dynamics can be generated by such simple logical structures as a single (full and ambiguous) circuit. PMID- 12754939 TI - Hardware (DNA) circuits. AB - A scheme is presented whereby a new genetic control circuit can be introduced into an organism, permitting the experimenter to turn the expression of a given gene (or set of genes) on or off at will. The proposed scheme involves a positive feedback loop--here, a positive regulator, the CII protein of phage lambda, with its structural gene engineered so as to require CII for its expression. This feedback loop creates the possibility of two stable steady states, with gene cII ON or OFF. Genes added downstream of cII and lacking a promoter will follow the same expression as cII. Two additional circuits allow the experimenter to switch at will between the ON and OFF states. PMID- 12754940 TI - Separating objects and 'neural' computation. AB - This article demonstrates how that the effectiveness of nervous system in doing the computations essential to an organism can be based on using algorithms that are readily implemented by nervous system 'device biophysics'. Collective effects and collective algorithms that exploit their dynamics provide powerful potential for useful neuronal computations. PMID- 12754941 TI - Complex pattern formation by a self-destabilization of established patterns: chemotactic orientation and phyllotaxis as examples. AB - Stable patterns can be generated by molecular interactions involving local self enhancement and long-range inhibition. In contrast, highly dynamic patterns result if the maxima, generated in this way, become destabilized by a second antagonistic reaction. The latter must act local and must be long-lasting. Maxima either disappear and reappear at displaced positions or they move over the field as travelling waves. The wave can have unusual properties in that they can penetrate each other without annihilation. The resulting pattern corresponds to those observed in diverse biological systems. In the chemotactic orientation of cells, the temporary signals allow the localized extensions of protrusions under control of minute external asymmetries imposed by the chemoattractant. In phyllotaxis, these signals lead to successive leaf initiation, whereby the longer lasting extinguishing reaction can cause a displacement of the subsequent leaf initiation site by the typical 137.5 degrees, the golden angle. On seashells, this patterns leads either to oblique lines that can cross each other or to oblique rows of dots. For some of the models animated simulations are available at http://www.eb.tuebingen.mpg.de/abt.4/meinhardt/theory.html. PMID- 12754942 TI - On the mechanochemical theory of biological pattern formation with application to vasculogenesis. AB - We first describe the Murray-Oster mechanical theory of pattern formation, the biological basis of which is experimentally well documented. The model quantifies the interaction of cells and the extracellular matrix via the cell-generated forces. The model framework is described in quantitative detail. Vascular endothelial cells, when cultured on gelled basement membrane matrix, rapidly aggregate into clusters while deforming the matrix into a network of cord-like structures tessellating the planar culture. We apply the mechanical theory of pattern formation to this culture system and show that neither strain-biased anisotropic cell traction nor cell migration are necessary for pattern formation: isotropic, strain-stimulated cell traction is sufficient to form the observed patterns. Predictions from the model were confirmed experimentally. PMID- 12754943 TI - [Mental health in a municipality of the State of Queretaro: a community-based psychiatric research model]. AB - The main object of this study was to determine, by means of a house-to-house survey, prevalence of mental disorders and type of alcohol consumption in an underprivileged population from a municipality in the State of Queretaro, Mexico. Results show that there was an 18.26% prevalence of psychiatric disorders with psychiatric comorbility of 56.8%. Anxiety disorders with 14.9% were the most frequent diagnosis and additional problems in descending order were affective disorders (10.2%), alcohol abuse and dependency (4.9%), schizophrenia (2%), and drug abuse (1.2%). Total prevalence was above that reported previously in Mexican population. There was a 48.8% association between unemployment and presence of one or more psychiatric disorders. Nearly 50% of male population had a problem with or excessive consumption of alcohol. These results lead us to consider that this is a high-risk population and that efforts must be made to continue these assessments to better determine prevalence of mental disorders and problems associated with them as well as the optimal mechanisms for attention to therm. PMID- 12754944 TI - [Use of base deficit as a prognosis factor for acute pancreatitis]. AB - INTRODUCTION: Acute pancreatitis (AP) severe is accompanied by important morbidity and mortality. Tissular hypoperfusion has been suggested as a severity determinant. Base deficit (BD) demonstrated to be a good indicator of hypo perfusion. We established predictive value in AP. MATERIAL AND METHODS: A retrospective, longitudinal, descriptive, and observational study was carried out. We included hospitalized patients from January 1996 to December 2000 with confirmed diagnosis of AP for laboratory, tomography and/or surgery; and with determination of BD on admission. Patients were divided into groups with or without BD and were compared with certain severity and mortality. RESULTS: We study 104 patients, 40(38%) without BD and 64 (62%) with BD. The mortality belonged to 22 patients (21.2%), all of the group with BD. The BD had a sensibility of 71.4% to predict severity and of 100% for the motility DISCUSSION: BD allows predicting severity and mortality in AP. These points out the role that carried out tissular hip perfusion; in addition, it can serve as guide for appropriate treatment. PMID- 12754945 TI - [Family-physician factors associated with glycemic control of patients with diabetes mellitus]. AB - INTRODUCTION: Metabolic control of patients with diabetes is determined by several factors, among others competence level, performance, and attitudes of primary-care physicians. OBJECTIVE: On objective was to determinate family physician factors associated with glycemic control in subjects with diabetes in two Family Medicine Units (FMU). DESIGN: Cross-sectional comparative study. SETTING: Two Family Medicine Units of the Mexican Institute Social Security. INTERVENTIONS: Family physician demographic factors were measured (age, sex, job seniority, time of university graduation, and had to have studied a specialty residence studied). Two validated instruments were applied to measure attitude level and handling competence of DM 2, and we determined glycemic level in the previous months of five diabetic patients by consulting room. RESULTS: Forty family physicians were evaluated, mean age 43.9 years, 57.5% with a residence in Family Medicine. Better glycemic control was associated with job seniority, OR 2.49 (CI 0.96-6.6), time of university graduation > 10 years with OR 2.11 (CI 1.4 2.9), to have at least one course related with diabetes in the previous year with OR 4.8 (IC 0.39-22), and competence level OR 2.02 (CI 0.36-11.3) CONCLUSIONS: There association between better glycemic control and more professional experience and training on diabetes in the previous year. PMID- 12754947 TI - [Diagnostic value of gamma-glutamyltransferase in the metastatic disease of patients with renal cell carcinoma]. AB - OBJECTIVE: Our objective was to determine the diagnostic value of gamma glutamyltransferase (GGT) as a marker of metastatic disease in patients with renal cell carcinoma (CCR). MATERIAL AND METHODS: A study of diagnostic tests was carried out to determine the value of GGT as a marker of metastatic disease in patients with neoplasms of kidney cells (NKC). Eighteen patients from the Urology Service at the Specialties Hospital, National Medical Center La Raza were included in the period from March to December 1997 with diagnosis of NKC, independently of co-morbid states. RESULTS: In an independent and blinded form, studies of tumorlike extension were carried out on patients (bone gammagram, axial computered tomography of abdomen, thorax and skull) to determine whether localized NKC or metastatic NKC took place. We obtained sensitivity of 71.42%, specificity of 90.90%, positive predictive value 83.33%, and negative predictive value, 83.33%. CONCLUSIONS: The study demonstrated that GGT values are high in a great percentage of patients with metastatic NKC, whereas in patients with localized CCR this enzyme practically is within normal ranges. Thus we are able to consider that GGT is useful as an indicator of metastasic disease in patients with CCR. PMID- 12754946 TI - [Mycoses frequency in three communities in the North mountain of the State of Puebla]. AB - In order to know mycosis frequency in the North of the State of Puebla, Mexico, in habitants from the communities of Ayotoxco, Mazatepec and Zacatipan were studied. Previous medical study biological samples were submitted to direct examination, smear and culture. Histoplasmin and sporotrichin skin test were applied to 57 individual from Zacatipan. From 110 patients 146 mycological studies were performed. Eighty six cases (59%) of mycosis were detected: 43 finger or toenails onychomycosis, 25 tinea pedis, seven tinea capitis, four cases of tinea manum and, finally, five cases of seborrhoeic dermatitis and two of pitiriasis versicolor. We isolated: 18 streins of dermatophytes, mainly Trichophyton rubrum and T. mentagrophytes (11 and 5 strains respectively); 12 cultures of non-dermatophytes filamentous fungi; six cases of mycelia sterile; six yeast strains, most of them Candida spp but none C. albicans. From 57 patients to whom skin tests were applied, five of them (8.8%) were positive to both antigens; ten positive (17.6%) only to histoplasmin and eight (14%) to sporotrichin. This study showed that rural population from Puebla present a high frequency of superficial mycosis (61% of mycological studies). Considering the percentage of positive skin test we suppose that there are many not diagnosed sporotrichosis and histoplasmosis cases. PMID- 12754948 TI - [Receptors and functions of TGF-beta, a crucial cytokine in wound healing]. PMID- 12754949 TI - [Postoperative entero-cutaneous fistula]. AB - Enterocutaneous fistulas of the digestive tract is a major catastrophe of surgical practice. In most cases, they represent serious complications of abdominal surgery. A total of 90% of cases develop of a surgical complication or injury. Despite progress in the management of fistulas, they were traditionally associated with high morbidity and mortality rates. The three major complications of fistulas have been electrolyte disturbance, malnutrition and sepsis. Complications are strongly related to anatomic site of fistula, to biochemical and electrolyte content of discharge, output, and underlying pathology. The ultimate objective in management of patients with enterocutaneous fistulas is fistula closure. In 1964, Chapman proposed management for fistulas in which a set of priorities in treatment was emphasized. Medical treatment is intended to cure (spontaneous closure) or to prepare patients for surgery. It was recognized that adequate nutritional support is an essential part of treatment of enterocutaneous fistulas. Control of sepsis is a priority because sepsis is the most common cause of death. Uncontrolled sepsis should be attacked as early as possible. Although spontaneous closure has increased in most series, it is likely that the majority of patients will require an operation and this should be performed at the proper time. PMID- 12754950 TI - [Hepatic insufficiency syndrome in a newborn]. PMID- 12754951 TI - [Uterine rupture in the second trimester of an interstitial ectopic pregnancy]. PMID- 12754952 TI - [Sarcoidosis and brucellosis: a strange and infrequent association]. AB - A cattlewoman, a 63-year-old patient with sarcoidosis associated to mellitensis brucellosis, was studied. Clinical manifestations of both multisystemic diseases were compared, as well as diagnostic and therapeutic aspects; we concluded that coexistence of both entities is infrequent but may occur due to increased presentation of the diseases. PMID- 12754953 TI - [Laennec and the creation of auscultation signs]. AB - Toward the end of the 18th century, clinical diagnosis in medicine shifted its focus from reliance on symptoms, which translates to subjective experience of illness, to signs, objective manifestations of pathologic changes. Several techniques were developed to elicit signs in clinical practice, and Laennec used them routinely. He palpated and prodded his patients to get an idea of changes in internal organs. He also applied his ear directly to his patient's chest to hear their heartbeat. On one occassion, he was unable to use these techniques and had the happy occurrence of rolling up a notebook to hear his patient's chest. This led him to hear a great number of new sounds. Through detailed observations, he was able to describe, classify, and correlate these sounds with autopsy findings, thus creating a new semiology of chest diseases. In this essay explore how in which Laennec created his instrument and system of signs of chest diseases, and how he was able to transmit his inventions to his colleagues. PMID- 12754954 TI - [Appendicular mucocele. Ultrasound finding]. PMID- 12754955 TI - [The double-helix 50th anniversary]. PMID- 12754956 TI - [What clinicians should know about human papilloma virus vaccines]. AB - Human papillomaviruses (HPVs) are epitheliotropic viruses that infect the basal layer of mucosal and keratinizing epithelia. HPV viral genome is made up of an early transcription region (E) and a late region composed of genes L1, L2, and a long control region (LCR). Despite the benign character of most lesions, HPV oncogenicity has been demonstrated in anal cancer, epidermodysplasia verruciformis, and cervical cancer. Nearly 12% of worldwide cancer incidence is due to HPV infection and HPV-16 is the most prevalent genotype found. Therefore, efforts in vaccines against HPVs have been directed mainly toward this genotype to dramatically diminish worldwide anogenital cancer incidence. Therapeutic vaccines are based on induction of an immunologic response against infected cells that express modified viral antigens E6 and E7. Prophylactic vaccines are based on prevention of infection by means of induction of humoral immunity against capsid viral proteins L1 and L2. This article reviews basics of the design of HPV vaccines and the type of vaccines currently being evaluated in clinical studies. PMID- 12754957 TI - [Ethic and legal aspects of informed consent in medical practice and research]. PMID- 12754958 TI - [Current medical practice. Pressures, depressions, and illusions]. PMID- 12754959 TI - [Symptoms and creativity. Frederick Chopin's case]. PMID- 12754961 TI - Perspectives. Competitive Medicare: different models serve different goals. PMID- 12754960 TI - [Of death, autopsies and respect]. PMID- 12754962 TI - [Physiopathology and etiology of mitral insufficiency]. AB - Mitral regurgitation has a complex pathophysiology. It should be assessed from the study of factors influencing regurgitant volume and the evaluation of hemodynamics effects downstream (impact on left ventricular function) and upstream (level of left atrial compliance and pulmonary pressure). The regurgitant volume is larger when the regurgitation time is longer, the regurgitant orifice is bigger and the magnitude of the left ventrico-atrial systolic gradient higher. The study of left ventricular function is difficult, especially in chronic mitral regurgitation where the apparently normal left ventricular systolic function can hide a significant worsening in myocardiacs fibres contractile abilities. With the increase in life expectancy and with the decrease in the incidence of rheumatic fever, aetiologies of mitral regurgitation have changed in the past 30 years. They are now dominated by dystrophic mitral regurgitation and infective endocarditis while rheumatic fever becomes less frequent. PMID- 12754963 TI - [Use of echocardiography in mitral regurgitation for the assessment of its mechanism and etiology for the morphological analysis of the mitral valve]. AB - Echocardiographic assessment of mitral regurgitation allows the diagnosis of its mechanism and cause which are major determinants in the feasibility of mitral valve repair. This assessment is based on a systematic analysis of the different structures of the mitral valve apparatus: mitral annulus (enlargement, calcification), mitral valve morphology (thickening, calcification, floppy valve, vegetations, perforation), mitral valve motion (restriction, identification of the prolapsed leaflets and scallops in patients with mitral valve prolapse or flail leaflets), subvalvular apparatus (ruptured chordae, thickening), papillary muscles, and left ventricular wall. This analysis can diagnose the mechanism of mitral regurgitation according to the Carpentier classification, and can clarify its cause: degenerative lesions (prolapse or flail leaflet with or without ruptured chordae), rheumatic lesions (thickened valves with restricted motion), endocarditis (vegetations, perforation, ruptured chordae), ischemic mitral regurgitation (restricted valve motion with inferior or posterior left ventricular wall asynergy), or functional mitral regurgitation (annular dilatation, displacement of papillary muscles with restricted leaflet motion). Transthoracic echocardiography with harmonic imaging usually allows a comprehensive assessment of functional anatomy of mitral regurgitation. Transesophageal echocardiography is indicated if transthoracic echocardiography is inadequate. It is also indicated just before surgery and as an intraoperative procedure. Real time 3D echocardiography should probably complete the evaluation of mitral regurgitation in the near future. PMID- 12754964 TI - [Quantification of mitral regurgitation]. AB - The echographic methods of quantification of mitral regurgitation are various. Semiquantitative methods using the color Doppler extension of the regurgitant jet are now replaced by more quantitative methods, including PISA, jet width, and regurgitant fraction. Although sometimes difficult, accurate quantification of mitral regurgitation is now possible in a majority of patients using transthoracic echocardiography. PMID- 12754965 TI - [Determinants and prognosis of ischemic mitral regurgitation]. AB - Ischemic mitral regurgitation (IMR) is mitral regurgitation (MR) due to complications of coronary artery disease. Two mechanisms can be individualized. Acute MR secondary to ruptured papillary muscle is a rare but often fatal complication of myocardial infarction. We focus on functional MR, much more common, which occurs without any intrinsic valve disease. It was often underrated because of low murmur intensity but is observed between 15 and 20% after a myocardial infarction. The presence and degree of the regurgitation are related to local left ventricular remodeling. The apical and posterior displacement of papillary muscles leads to excess valvular tenting which in turn, in association with loss of systolic annular contraction, determines the severity of the regurgitation. IMR presence is associated with an excess mortality. The mortality risk is directly related to the degree of the regurgitation and a regurgitant volume > or = 30 ml or an effective regurgitant orifice > or = 20 mm2 define a high-risk group. In current clinical practice, IMR is mainly corrected with ring annuloplasty. However, this technique does not correct local alterations of left ventricular remodeling and its benefits on long-term outcome remains to be demonstrated. PMID- 12754966 TI - [Mitral regurgitation in infective endocarditis]. AB - Endocarditis affecting the mitral valve is frequent and is associated with specific features. Mitral prolapse is the most frequent underlying disease and the mechanism of mitral regurgitation secondary to infective endocarditis is frequently complex and multiple. Echocardiography plays a key-role in both the diagnosis, the prognostic assessment and the choice of the best therapeutic option in patients with mitral valve endocarditis. Surgery is frequently necessary, and must be performed early in the course of the disease. Mitral valve repair is the best therapeutic option, when technically possible. PMID- 12754967 TI - [Prognosis of organic mitral regurgitation and implications for surgical indications]. AB - The term organic Mitral Regurgitation (MR) relates to MR secondary to anatomic alteration of the valvular or subvalvular mitral apparatus and refers to rheumatic MR and degenerative MR, i.e. mitral valve prolapse, which has become in the past 20 years the 1st cause of severe MR leading to surgery in western countries. Recent publications on the prognosis of patients with MR secondary to flail leaflet, showed that these patients incur excess mortality rates as compared to expected and that ten years after diagnosis, 90% of those will either be dead or operated on for severe symptoms. On the other hand, analysis of postoperative prognosis showed that the best results of surgical correction were observed in asymptomatic patients with normal pre-operative left ventricular function. The prognosis of these patients was then similar to that expected if a valvular repair was performed, making of mitral repair the hinge point of early surgical strategies. PMID- 12754968 TI - [Surgical treatment of mitral valve regurgitation: which method for which patient?]. AB - The surgical treatment of mitral regurgitation has been completely revamped in the last years. The development of reproducible and durable plasty techniques has made mitral valve replacement less and less frequent. The virtual disappearance of rheumatic valve disease in developed countries has made degenerative disease the most frequent etiology of mitral insufficiency. Furthermore the outlook of this pathology has been modified by the early surgical treatment. The following article presents the surgical techniques available today and their indications in the different types of mitral insufficiency. PMID- 12754969 TI - [Usefulness of intraoperative echocardiography for the management of mitral regurgitation]. AB - Intraoperative transesophageal echocardiography is crucial in the management of patients with mitral regurgitation. In fact, a vast majority mitral insufficiencies may actually be repaired. This kind of surgery require an excellent definition of anatomic lesions and mechanisms of mitral regurgitation in order to optimize the result of the repair. In this field, intraoperative echocardiographic findings are complementary with anatomical findings for the surgeon. Before surgery, intraoperative echography help to describe all valvular lesions and mechanisms of the regurgitation. Three-dimensional echocardiography may be obtained using intraoperative echography and may be useful in mitral valve prolapse to better define the location and size of prolapse and to communicate informations to the surgeon. After repair, echographyallow the control of the result and also of right and left ventricular function. In case of major abnormality such as residual significant mitral regurgitation or important intra ventricular obstruction, a second procedure may be required. The collaboration between the echographist and the surgeon is crucial to obtain the best results in mitral valve repair. Indications for intraoperative echocardiography are dependent on the experience of the surgeon and the complexity of mitral lesions to repair. PMID- 12754970 TI - [Mitral stenosis]. AB - The decrease in the incidence of acute rheumatic fever in western countries has led to a sharp decrease in the incidence of mitral stenosis. This decrease also modifies the clinical presentation and mitral stenosis is now encountered in older patients who have severe impairment of valve anatomy. In developing countries, mitral stenosis remains a frequent disease. The management of patients with mitral stenosis has been modified by the development of percutaneous mitral commissurotomy whose safety and efficacy have been demonstrated in a number of studies with a follow-up of up to ten years. Percutaneous mitral commissurotomy is now the reference treatment for mitral stenosis with pliable valves in young patients and its efficacy has been validated in randomised trials versus surgery. Mitral stenosis in older patients, as it is encountered in western countries, is a more heterogeneous group. Predictive analysis has shown that the predictions of immediate and late results are multifactorial. This has led to consider the indications for percutaneous mitral commissurotomy in patients who do not have ideal anatomic conditions, when their other characteristics are favourable. This is particularly the case in young patients who do not have a very tight mitral stenosis (1-1.5 cm2) and who do not have an advanced heart disease. When the conditions are favourable, percutaneous mitral commissurotomy can be considered in patients who have few symptoms, in particular in order to reduce the thromboembolic risk. PMID- 12754971 TI - [Tranexamic acid versus aprotinin for the prevention of bleeding during mitral valve surgery ]. AB - PURPOSE: This retrospective stady has for objective to compare the effect of Tranexamic Acid (TA) to the low dose of aprotinin (AP) in primary mitral valve surgery in terms of blood loss and transfusion requirements. METHODS: Are included in the study operated patients of a valvulopathy mitral isolated. Two groups of 50 patients are collected. The tranexamic acid group has received 30 mg kg-1 the acid tranexamic and the aprotinin group has received a low regimen as 500,000 UIK of aprotinin. Blood loss by the chest drains are assessed to different times during first 24 hours post cardiopulmonary bypass. In the same way, we have measured the platelet and fibrinogen count. Blood products were administered according to a classic protocol. RESULTS: The two groups are comparable clinic and echocardiographic parameters what authorizes us an appariement acceptable. Various cardiopulmonary bypass times are almost similar. We noticed a tendency to excessive blood loss processed by low regimen aprotinin and a significant rate difference of platelet and the fibrinogen level. But no complication has been recorded in the two groups. CONCLUSIONS: This study demonstrates relatively different effect of the two fibrinolytics inhibitors in primary mitral valve surgery. As for the superiority of one of the two produces, it needs a confirmation by a randomised and controlled clinical trial. PMID- 12754972 TI - [Huge right atrial myxoma revealed by heart failure]. AB - We report a case of 34 years old female with a diagnosis of a big right Atrial myxoma revealed by echocardiography. The treatment consisted on a surgical excision of the tumour under extra corporal circulation. The patient made a good post-operative recovery. Authors showed the clinical signs of these tumours to be various, and echocardiography to be the definitive diagnosis procedure. PMID- 12754973 TI - Pain control needs sometimes overlooked. PMID- 12754974 TI - More advice for chronic skin conditions. PMID- 12754975 TI - Drug companies respond to NP boycott threat. PMID- 12754976 TI - Conflict driving you crazy? PMID- 12754977 TI - Playing sports after a toe fracture. PMID- 12754978 TI - HIPAA's new privacy rules. PMID- 12754979 TI - Testing for STDs. PMID- 12754980 TI - Current management of diabetes mellitus in adults. Risk factor reduction is key. PMID- 12754981 TI - Patient information. Managing your high blood pressure. PMID- 12754982 TI - Dysesthetic vulvodynia. Management strategies to improve quality of life. PMID- 12754983 TI - Meditation as medicine. Benefits go beyond relaxation. PMID- 12754984 TI - Managing PMS and perimenopause symptoms. The role of compounded medications. PMID- 12754985 TI - Brewing up good health. Research says tea packs power. PMID- 12754986 TI - Pushing the edge of viability. Treatment dilemmas in neonatology. PMID- 12754987 TI - Selecting wound healing products. Choices for long-term care settings. PMID- 12754988 TI - A community-based approach to asthma care. The Durham County Asthma Coalition. PMID- 12754989 TI - Polymyalgia rheumatica. The great impressionist. PMID- 12754990 TI - Easing interstate practice. NPs get a turn at multistate licensure. PMID- 12754991 TI - [The pathophysiological mechanism of cardiac development and hypertrophy, and heart failure]. PMID- 12754992 TI - [The epidemiology of heart failure in Japan]. AB - Very limited epidemiological data are available regarding the prognosis of heart failure and temporal changes in survival in a population-based setting in Japan. It is estimated from the total sales of digitalis, its prescription rate and usage ratio for arrhythmias that at least one million of patients suffer from heart failure in Japan. Dilated cardiomyopathy (DCM) is a major cause of severe heart failure. Over the past 27 years, three cohorts corresponding to three different periods of 9 years were investigated. Ten-year survivals were progressively improved. Patients were older and less severely affected, partly explaining the improvement in survival. In the subgroups treated with ACE inhibitors and beta blockers, calculated ten-year survival rate reached over 90%. PMID- 12754993 TI - [Acute heart failure]. AB - Acute heart failure is a life-threatening medical emergency and appropriate management can reduce the morbidity and mortality of heart failure. Despite advances in treatments, the number of deaths has increased steadily. Therefore, the guideline was assigned to convey to the physicians the virtue of medical management with recent trends in pharmacological and nonpharmacological treatments. Japanese new guidelines for the management of acute heart failure have been published from the Japanese Circulation Society in October, 2000, which was partially based on ACC/AHA guideline in 1995 but prepared under consideration of medical benefits system and health care system in Japan. We expect that the guideline would be revised to make it more useful with continual advances in the management strategies in the future. PMID- 12754994 TI - [Guidelines for the evaluation and management of chronic heart failure]. AB - The Japanese Circulation Society have long been involved in the development of practice guidelines designed to assist physicians in the management of chronic heart failure. Importantly, as our population ages, the incidence of chronic heart failure and its mortality rate will continue to increase. Furthermore, recent advances in our understanding of the pathophysiology of chronic heart failure and new developments in the therapy of this disorder have greatly expanded the information base which to make decisions. The guidelines for the evaluation and management of chronic heart failure were developed by cardiovascular specialists and were based primarily on a comprehensive review of published reports. Furthermore, these guidelines are based on the assumption that the resources necessary to provide this care readily available. PMID- 12754995 TI - [Phenotype alterations of failing myocardium]. AB - Compared with the mechanisms of adaptive responses of the heart, the mechanisms of maladaptation have been less elucidated. However, the followings are probably responsible for the progression of heart failure: 1) progressive loss of cardiac myocytes (quantity change); 2) progression of systolic dysfunction of individual myocytes(quality change); or 3) both. Apoptosis may be responsible for the myocyte death although controversial. Myocytes of failing hearts shows dramatic phenotype alterations in the structure, function, and biochemical events such as fetal type gene induction. Hypertrophy and degeneration of myocytes are the common, but not specific, features in failing hearts. Meanwhile, not only myocyte phenotype alterations but also myocardial interstitial changes such as extracellular matrix deposition, activation of fibroblasts, and narrowing of vessel lumens play important roles for the progression of heart failure. PMID- 12754996 TI - [Alterations in sympathoadrenal system in congestive heart failure]. AB - Although sympathoadrenal system has a pivotal role in maintaining homeostasis during stress, protracted activation adversely affects life expectancy in patients with heart failure. Detrimental effects of sympathoadrenal activation over time have been confirmed using transgenic mice overexpressing beta 1 adrenergic receptors, which showed that initial hypercontractility led to cardiac hypertrophy with failure, resulting in premature death. Mechanisms underlying such adverse effects involve multiple biological events including production of oxygen free radicals, cytokines, matrix metalloproteinase, apoptosis, cardiac hypertrophy, and chamber remodeling as well as energy expenditure. Desensitization phenomenon of the beta-adrenergic receptors is one of the predominant features characterizing congestive heart failure. Down-regulation of the receptors, increases in inhibitory guanine-nucleotide(G) protein and G protein-coupled receptor kinases are responsible for the phenomenon. PMID- 12754997 TI - [Roles of cytokines in the pathogenesis of heart failure]. AB - Cytokines are being increasingly recognized as important factors in the pathogenesis and pathophysiology of heart failure. Elevated levels of circulating cytokines have been reported in patients with heart failure, and various cytokines have been shown to depress myocardial contractility in vitro and in vivo. Our recent study showed that the various drugs for heart failure modulated the production of cytokines, and some of these drugs inhibited activation of NF kappa B. Cytokine gene therapy which inhibits inflammatory response by viral IL 10 and IL-1 receptor antagonist has been shown to be effective in the animal models of heart failure. Mast cells have been shown to play important role in the pathogenesis of heart failure due to viral myocarditis, and transition from compensated hypertrophy to heart failure. PMID- 12754998 TI - [Renin-angiotensin system and heart failure]. AB - The local renin-angiotensin system in cardiac tissue has been demonstrated to be involved in the pathophysiology of cardiac hypertrophy and heart failure. Angiotensin II type 1(AT1) receptor stimulation leads to vasoconstriction, cell growth, oxidation, aldosterone release, and fibrosis. Recent evidence suggests that angiotensin II type 2(AT2) receptor exerts cardioprotective effects by counteracting against AT1 receptor. However, the role of AT2 receptor signaling in cardiac hypertrophy and fibrosis still remain controversial. This review focused on the accumulating evidence for the cardiac renin-angiotensin system in heart failure. PMID- 12754999 TI - [Role of oxidative stress in heart failure]. AB - Recent experimental and clinical studies have suggested that oxidative stress is enhanced in heart failure. The production of oxygen radicals is increased in the failing heart whereas antioxidant enzyme activities are preserved normal. Mitochondrial electron transport is an enzymatic source of oxygen radical generation and also a target against oxidant-induced damage. Chronic increases in oxygen radical production in the mitochondria can lead to a catastrophic cycle of mitochondrial DNA damage as well as functional decline, further radical generation, and cellular injury. These cellular events might play an important role in the development and progression of myocardial remodeling and failure. PMID- 12755000 TI - [Myocyte renewal]. AB - In contrast to conventional assumption that myocytes are never renewed after birth, a growing body of evidence suggests that human cardiac myocytes might divide in myocardial infarction and severe heart failure. Bone marrow cells may also contribute to myocyte regeneration, when injected or mobilized into systemic circulation by cytokines. A clinical study demonstrated that intracoronary administration of autologous bone marrow significantly improved the cardiac function after acute myocardial infarction. No adverse effect was found. Cell therapy using adult stem cells is anticipated to be an effective treatment of heart failure. PMID- 12755001 TI - [Membrane-anchored heparin-binding EGF-like growth factor processing by ADAM12 in cardiac hypertrophy]. AB - G-protein coupled receptor(GPCR) agonists are well-known inducers of cardiac hypertrophy. We found that the shedding of HB-EGF via metalloproteinase activation and subsequent transactivation of the epidermal growth factor receptor occurred when cardiomyocytes were stimulated by GPCR agonists, leading to cardiac hypertrophy. A new inhibitor of HB-EGF shedding, KB-R7785, blocked this signaling. We cloned a disintegrin and metalloprotease 12(ADAM12) as a specific enzyme to shed HB-EGF in the heart and found that dominant negative expression of ADAM12 abrogated this signaling. KB-R7785 bound directly to ADAM12, suggesting that inhibition of ADAM12 blocked the shedding of HB-EGF. In mice with cardiac hypertrophy, KB-R7785 inhibited the shedding of HB-EGF and attenuated hypertrophic changes. These data suggest that shedding of HB-EGF by ADAM12 plays an important role in cardiac hypertrophy, and that inhibition of HB-EGF shedding could be a potent therapeutic strategy for cardiac hypertrophy. PMID- 12755002 TI - [Diagnosis by new echocardiographic techniques]. AB - Echocardiography has been very powerful for not only evaluating hemodynamics and cardiac function but also finding causes of heart failure. Moreover, recent advancements in echocardiography enable us to assess patients with heart failure more conveniently and precisely. The quality of 2-dimensional echocardiographic images is dramatically improved by introduction of harmonic imaging method and intravenous contrast agents which opacify the left ventricular cavity. Tissue Doppler echocardiography can provide information on regional myocardial velocity thereby enabling assessment of regional as well as global left ventricular function. Strain and strain rate of the regional myocardium can be also determined by the newest technology based on tissue Doppler echocardiography. PMID- 12755003 TI - [Plasma brain natriuretic peptide as a useful biochemical marker of congestive heart failure]. AB - Neurohumoral factors, which is divided to two groups, are activated in patients with congestive heart failure(CHF). One is cardiotoxic biochemical markers such as nor-epinephrine, angiotensin II, endothelin-1, and vasopressin, the other is cardioprotective biochemical markers such as atrial natriuretic peptide, and brain natriuretic peptide (BNP). Among various neurohumoral factors, plasma level of BNP is the most useful marker to diagnose and evaluate the severity and prognosis of CHF patients. The reason why the usefulness of plasma BNP is 1) BNP is of ventricular origin, 2) BNP reflects hemodynamic abnormalities(high left ventricular end-diastolic pressure and/or low left ventricular ejection fraction), 3) Attenuation of biological compensation of beneficial effects of BNP(vasodilation, diuresis) in advanced-staged CHF. PMID- 12755004 TI - [Chronic heart failure assessed by nuclear medicine]. AB - Radionuclide imaging has been used for comprehensive evaluation of patients with heart failure. First of all, radionuclice ventriculography is useful for objective and quantitative assessment of LV systolic function. This is particularly important for assessment of severity of the disease and treatment effects. Radionuclide myocardial imaging permits evaluation of cellular and molecular function of the heart failure, including myocardial viability on the basis of membrane integration or preservation of myocardial metabolism. The combination of oxidative metabolism and cardiac workload may provide objective assessment of myocardial energy efficiency. Furthermore, recent developments of new radiopharmaceuticals permit noninvasive assessment of adrenergic neuronal function and beta-receptor function in vivo. The severity of the adrenergic dysfunction in patients with heart failure may be related the outcome of these patients. Such analysis may be valuable for evaluation of treatment effects and may possibly provide the best treatment for each patient with heart failure. PMID- 12755005 TI - [Clinical application of exercise testing in heart failure]. AB - Patients with heart failure frequently complain of fatigue and/or dyspnea during daily life. These exertional symptoms can be evaluated by the cardiopulmonary exercise testing. Peak oxygen uptake, anaerobic threshold(AT), the ratio of the increase in minute ventilation to the increase in carbon dioxide output(VE-VCO2 slope), the slope of the increase in oxygen uptake relative to the increase in work rate (delta VO2/delta WR), and the time constant of oxygen uptake (tau on) are reported to be useful to assess the severity and prognosis of heart failure patient. The information obtained from cardiopulmonary exercise testing can be used to select therapeutic option to improve both functional capacity and prognosis, and to identify patients with the greatest need for cardiac transplantation. PMID- 12755006 TI - [Treatment of heart failure based on large-scale clinical trials: renin angiotensin system antagonists and beta-blockers]. AB - Recent large-scale clinical trials have provided credible evidence that angiotensin-converting enzyme inhibitors and beta-blockers can prolong the survival in patients with chronic heart failure. Treatment of asymptomatic left ventricular dysfunction is as important as treatment of symptomatic disease. Addition of aldosterone antagonists to angiotensin-converting enzyme inhibitors and beta-blockers may afford further benefits in patients with severe heart failure. If angiotensin-converting enzyme inhibitors or beta-blockers are not tolerated, angiotensin receptor blockers should be considered as the alternative. The present article will review recent advances in treatment of heart failure based on large-scale clinical trials. PMID- 12755007 TI - [Inotropic agents]. AB - Depression of myocardial contractility plays an important role in the development of heart failure and many inotropic agents were developed to improve the contractile function of the failing heart. Agents that increase cyclic AMP, either by increasing its synthesis or reducing its degradation, exerted dramatic short-term hemodynamic benefits, but these acute effects were not extrapolated into long-term improvement of the clinical outcome of heart failure patients. Administration of these agents to an energy starved failing heart would be expected to increase myocardial energy use and could accelerate disease progression. The role of digitalis in the management of heart failure has been controversial, however, the recent large scale clinical trial has ironically proved that digoxin reduced the rate of hospitalization both overall and for worsening heart failure. More recently, attention was paid to other inotropic agents that have a complex and diversified mechanism. These agents have some phosphodiesterase-inhibitory action but also possess additional effects, including cytokine inhibitors, immunomodulators, or calcium sensitizers. In the Western Societies these agents were again shown to increase mortality of patients with severe heart failure in a dose dependent manner with the long-term administration. However, it may not be the case in the Japanese population in whom mortality is relatively low. Chronic treatment with inotropic agent may be justified in Japanese, as it allows optimal care in the context of relief of symptoms and an improved quality of life. Therefore, each racial group should obtain specific evidence aimed at developing its own guidelines for therapy rather than translating major guidelines developed for other populations. PMID- 12755008 TI - [Use of beta-blockers in heart failure]. AB - Recently, a number of clinical trials showed that beta-blockers reduce morbidity and mortality in patients with heart failure. Possible mechanisms for this effect of beta-blockers include protection from catecholamine myocyte toxicity, upregulation of beta-adrenergic signaling, attenuation of apoptosis, reduction in myocardial oxygen demand, and antiarrhythmic effect. Metoprolol, bisoprolol, and carvedilol are proved to have substantial benefit in the treatment of heart failure by large-scale studies. In patients with moderate to severe heart failure, these drugs have to be started slowly and to be uptitrated gradually while watching for deterioration of heart failure. Cardiac contraindications include severe bradycardia, high-degree heart block, and decompensated heart failure. In spite of those adverse effects, beta-blocker should be actively adopted in the treatment of heart failure. PMID- 12755009 TI - [Renin-angiotensin system as a potential target for the treatment with heart failure: ACE inhibitors, angiotensin-II receptor blockers and aldosterone antagonists]. AB - Recent experimental studies and clinical trials have revealed that the modulation of either systemic or regional(cardiovascular) renin-angiotensin systems(RAS) is one of the potential targets to prevent the progression of heart failure. Either ACE inhibitor, angiotensin-II receptor blocker or aldosterone antagonist differently block RAS. Many clinical trials told us that each of them improves heart failure and might promise the better prognosis. Now, some experimental studies are encouraging us to test the comparative effects or the effects in combination of them. Here we will summarize the current outcomes from the experimental and clinical studies, and discuss the future direction. PMID- 12755010 TI - [Positive inotropic agents]. AB - Despite wide prevalence of beta-blockers and angiotensin-converting enzyme inhibitors for chronic heart failure, their therapeutic efficacy remains limited because effects of these drugs depend largely on pathophysiology of an individual patient. Especially, positive inotropic therapy could be deleterious for some patients, whereas effective in improving symptoms for some patients. Recent clinical experience has led to a concept that in order to be clinically beneficial and safe, positive inotropic agents should only enhance myocardial contractility to a very modest degree in moderately to severely symptomatic patients. Positive inotropic agents could eventually serve as an individualized therapeutic option in effective management of chronic heart failure. PMID- 12755011 TI - [Indications, management, and complications for the use of a percutaneous intra aortic balloon counterpulsation and cardiopulmonary support system]. AB - Development and improvement of mechanical cardiac support systems, for example, IABP, and PCPS, is so remarkable that recently, we can rescue severe heart failure, cardiogenic shock, and acute fulminant myocarditis with these mechanical support. Moreover, indications of these mechanical support systems are widened to the support of high risk coronary interventions such case as acute myocardial infarction with poor LV function. Mechanical cardiac support systems are very important for the treatment of high risk patients. In these mechanical support systems, percutaneous systems such as IABP and PCPS are widely used. So in this article, we review mainly indications, management, and complications of IABP and PCPS with acknowledgement of our hospital. PMID- 12755012 TI - [New surgical strategies for severe heart failure]. AB - Alternative surgical procedures to heart transplantation for patients with severe congestive heart failure are developing in the past decade. Left ventricular restoration such as partial left ventriculectomy, endo-ventricular circular patch plasty, or septal anterior ventricular exclusion is effective for patients with operative indication and functional mitral regurgitation due to dilated heart can be repaired effectively in some patients. It is important to understand the surgical procedures and the indications for not only cardiac surgeons but also cardiologists, and combination of surgical and medical treatment can be an alternative to heart transplantation for properly selected patients. PMID- 12755013 TI - [Cardiac transplantation]. AB - Cardiac transplantation has been established as a therapeutic strategy for patients with end-stage heart failure in the developed countries. In Japan, the first cardiac transplantation under the new legislation was successfully performed on February 1999 in Osaka University Hospital. Following this successful operation, the agreement with the transplantation under brain death seems to be further obtained in Japan. However, the number of the transplantation performed during the following 3 years was not enough. Thus, we wish that the number will further increase and that the transplantation will become one of the established therapeutic strategies for end-stage heart failure in our county. PMID- 12755014 TI - [Exercise therapy for chronic heart failure]. AB - Increased mortality and impaired exercise tolerance is the two major clinical problems in patients with chronic heart failure(CHF). In addition, for many years, bed rest has been recommended for patients with CHF. However, in the last decade, exercise training has been shown to improve exercise capacity, quality of life, and event-free survival in patients with CHF. Furthermore, exercise training improves autonomic nerve function and vascular endothelial function in these patients. Based on these lines of evidence, clinical practice guidelines of exercise therapy for CHF have recently been published. More effort should be made to establish the optimal exercise prescription and to increase the implementation of exercise training as a non-pharmacologic therapy for CHF. PMID- 12755015 TI - [Treatment for diastolic failure]. AB - Diastolic dysfunction is now recognized as an important mechanism of heart failure, but treatment for diastolic failure has not yet been established. Many of causes for diastolic dysfunction have been related to myocardial hypertrophy or fibrosis. Until recently, angiotensin II has been proposed as one of important growth factors that lead into hypertrophy and fibrosis. Angiotensin II blockade, therefore, is a promising candidate for the treatment of diastolic failure. Two randomized studies with angiotensin II type 1 receptor blocker are under way. This review will describe the pathophysiologic basis for diastolic dysfunction, and discuss the possible treatments for diastolic failure referring to the recently-proposed two guidelines. PMID- 12755016 TI - [Ischemic heart failure]. AB - Ischemic heart failure is induced by myocardial ischemia, which is probably the commonest cause of left ventricular systolic dysfunction. Theoretically, there are two main strategies to treat the patients with ischemic heart failure; to retard progression of the dysfunction, and to relief and prevent myocardial ischemia. The management with angiotensin converting enzyme(ACE) inhibitor and beta-blocker improve the prognosis of ischemic heart failure by enhancing the left ventricular function and preventing coronary event. Spironolactone also seems to improve their prognosis. In contrast, treatments directed specifically at coronary arteries, such as antithrombotic agents and revascularization, have not been proven to be effective and safe to these patients yet. Large scale clinical trials are currently underway investigating the effect of treatment, such as aspirin, warfarin, clopidogrel and revascularization which are targeted to the coronary syndrome. PMID- 12755017 TI - [Gene therapy for myocardial regeneration]. AB - Most of cardiac myocytes have lost their ability to proliferate and differentiate into new myocytes. Therefore, myocyte regeneration and replacement in adult myocardium was thought to be impossible by medical community. However, recent findings in cardiovascular researches provide the possibility that gene therapy and stem cell therapy could have supportive effects on myocyte regeneration and myocardial revascularization in damaged heart. Here, we presented recent progress, especially, in gene therapy for myocardial regeneration under ischemic heart disease and heart failure. PMID- 12755018 TI - [Angiogenic cell therapy]. AB - Therapeutic angiogenesis is an effective means for tissue salvage in patients with critical limb ischemia. Angiogenesis is defined as a formation of new blood vessels by sprouting of preexisting mature endothelial cells(ECs). In contrast, vasculogenesis is referred to as the creation of primordial blood vessels from endothelial progenitor cells (EPCs) or angioblasts. Neovascular formation in adults has been considered to result exclusively from the former process (i.e., angiogenesis). However, we and other researchers recently identified EPCs in human peripheral blood(PB), and circulating EPCs have been shown to accumulate at active angiogenic sites and to participate in neovascularization, a notion consistent with 'postnatal vasculogenesis'. EPCs in adults originate from bone marrow (BM), and we recently have demonstrated that in vivo implantation of autologous BM-MNCs effectively augmented ischemia-induced neovascularization in animal studies as well as human trial(TACT Trial). Here we summarize recent advances in cell transplantation-mediated therapeutic angiogenesis. PMID- 12755019 TI - [Artificial heart--turbo type blood pump for long-term use]. AB - Shortage of donor heart for transplantation necessitates long-term artificial assist heart. Turbo-pump is smaller, simpler and cheaper than the pulsatile displacement type pump, but the turbo-pump has defect of thrombus formation at the shaft seal. Our centrifugal pump with magnetically suspended impellers overcomes this defect and is ready for clinical trials now. The structures and functions are described and are compared with the other newly-developed pump of the same kinds with us. And also the pumps of centrifugal type and axial-type, of which impellers are supported by pivots, are reviewed briefly from the stand point for long-term use. Other pumps are referred too: pumps with hydrodynamic bearing and a pump with the shaft seal which is washed and cooled by saline solution. PMID- 12755020 TI - [Osteopontin and arthritis]. AB - Osteopontin is an RGDS-containing protein which acts as an intercellular signaling molecule as well as a cellular attachment protein. Expression of osteopontin has been observed in the joints of patients with rheumatoid arthritis as well as osteoarthritis. However, the functions of osteopontin in such diseased articular components have not yet been elucidated. Recent investigations using knockout mice lacking osteopontin indicated that the presence of osteopontin could play a role in the development of arthritis induced by injection with monoclonal antibodies raised against type II collagen. PMID- 12755021 TI - [Treatment of facial spasm using botulinum toxin]. PMID- 12755022 TI - [Development of a novel targeted therapy for human glioblastoma]. PMID- 12755023 TI - [Evaluation on major cerebral arterial stenotic lesions with three-dimensional magnetic resonance cisternograms and coordinated MR.CT angiograms]. AB - To evaluate major cerebral arterial stenotic lesions within a cisternal space, the outer wall contours of the arteries were depicted on three-dimensional (3D) MR cisternograms. The 3D MR cisternograms were reconstructed by perspective volume-rendering algorithm from the source volume data obtained from the T2 weighted 3D fast spin-echo sequence. Those images were compared with coordinated 3D MR angiograms, and then with 3D CT angiograms through the similar visual projections. The presence of stenotic lesions was indicated by the morphological discrepancy between the outer wall configuration of the itracisternal stenotic artery depicted on the 3D MR cisternograms and the intraluminal boundary shown on the 3D CT angiograms. With application of these techniques, spatial expansion of the stenotic lesions was able to be visualized. In this way, clinical 3D evaluation of the therapeutic effect on and follow-up of intracisternal major cerebral arterial stenotic lesions would be possible in patients with acute ischemic stroke. PMID- 12755024 TI - [Surgery for giant intracranial aneurysms using advanced technology]. AB - PURPOSE: Outcome of surgery for giant intracranial aneurysms is still unsatisfactory. The reason for complications is occlusion of perforators or parent arteries by the aneurysmal clipping itself or temporary occlusion of the main arteries. We report the surgical outcome of treatment of giant aneurysms using several advanced techniques which we devised to prevent these complications. MATERIALS AND METHODS: The subjects were eight patients with giant intracranial aneurysms who underwent surgery during the recent five years. Six patients had ruptured and two had unruptured aneurysms. Aneurysms were located at the ICA in five and the MCA in three patients. Aneurysmal sizes ranged from 25 to 50 mm. Preoperative 3DCTA was performed to investigate the aneurysm and the surrounding vessels in all cases. Patients with unruptured aneurysms at the ICA underwent balloon occlusion tests to check the potential for safe temporary occlusion of the parent artery, with SEP monitoring and Xe-SPECT. Intraoperative angiography and neuroendoscopes were used to prevent problems and complications which might be caused by aneurysmal clipping. RESULTS: In seven of eight cases, the aneurysmal neck was completely obliterated with clips and in one case the aneurysm was trapped with STA-MCA anastomosis. Glasgow Outcome Scale of the patients showed good recovery in six, moderately disabled (MD) in one and dead in one. The patient demonstrating MD developed hemiparesis due to vasospasm. One patient died from rebleeding of the aneurysm caused by slippage of the aneurysmal clip despite the confirmation of complete obliteration by intraoperative angiography. CONCLUSIONS: A better surgical outcome of treatment for giant aneurysms was obtained by temporary clips whose placement was based on the results of balloon occlusion test, as well as the use of intraoperative angiography and neuroendoscopes. PMID- 12755025 TI - [A ruptured choroidal artery aneurysm of the anterior inferior cerebellar artery obliterated via the endovascular approach: case report]. AB - Intraventricular aneurysms associated with fourth ventricular hemorrhage are rare. A case of a ruptured aneurysm in a choroidal branch of the right anterior inferior cerebellar artery (AICA) is reported here. A 56-year-old man presented with sudden onset of vertigo and nausea. CT scan showed an intraventricular hemorrhage within the fourth ventricle. Cerebellar angiography showed an aneurysm at the choroidal artery branching from the right AICA. The patient rejected both general anesthesia and craniotomy, so endovascular embolization under local anesthesia was performed using Guglielmi detachable coils (GDCs) and a fibered platinum coil. The distal portion of the right AICA and the aneurysm were obliterated. His postoperative course was fairly satisfactory. He suffered from a minimal gait disturbance caused by truncal ataxia for several days after the operation. He was discharged from hospital without neurological deficit. There have been only a few articles about choroidal artery aneurysms. As treatment, direct surgery has been recommended in past cases, but endovascular embolization of the parent artery was successfully performed in this case. Not only direct surgery but also endovascular surgery may be regarded as the treatments of choice for intraventricular aneurysms, depending on the size of the parent artery. PMID- 12755026 TI - [Two cases of impending herniation due to multiple traumatic acute subdural hematomas: combination of burr hole evacuation and craniotomy]. AB - A contralateral extra-axial hematoma sometimes occurs during an operation on an acute subdural hematoma and may become fatal. Using a combined procedure of burr hole evacuation and craniotomy, we treated 2 cases of multiple traumatic acute subdural hematomas. Our policy for such cases is first to perform a burr hole evacuation for the acute subdural hematoma in the emergency room, while simultaneously preparing the operation room for a possible further operation. Next, we perform computed tomography (CT) of the brain. If the evacuation does not provide enough decompression, we either carry out a craniotomy at the same site, or, we observe the patient without resorting to craniotomy. However, if the patient's condition deteriorates, burr hole evacuation is repeated and/or craniotomy is carried out as soon as possible on the lesion at the already prepared operation room. Both of our patients received craniotomy for another subdural hematoma after the burr hole evacuation. Though his intracranial pressure was well managed during the acute stage, one of the patients died 21 days after the trauma due to an extensive brain infarction caused by vasospasm. The other regained consciousness and was able to walk 5 months after the trauma in spite of cerebral infarction from vasospasm. The possible mechanism of vasospasm in severe head injury is also discussed. PMID- 12755027 TI - [A case of ruptured and unruptured developing cerebral aneurysms at the bilateral M2 bifurcations five years after surgery for ruptured aneurysm at the end of the azygos anterior cerebral artery]. AB - A 36-year-old woman with 4 cerebral aneurysms at unusual sites including bilateral M2 bifurcation aneurysms is reported. She had been in good health in the previous 5 years since the treatment for a ruptured aneurysm at the end of the azygos anterior cerebral artery when she was 31 years old. Five years later, she became comatose with a huge hematoma in the right temporal lobe due to the rupture of the aneurysm at the right M2 bifurcation, which had been very small 5 years ago. She underwent an emergent clipping operation, and then she became alert with motor weakness extremities of on her left-side. Postoperative angiograms revealed a de novo aneurysm at the left M2 bifurcation and an aneurysm at the origin of the lenticulostriate artery, which has remained unchanged for 5 years. An aneurysm at the M2 bifurcation is rare, especially when it is situated bilaterally at the mirror sites. To detect de novo aneurysms, postoperative angiographical follow-up should be considered in patients with multiple aneurysms and in young patients. PMID- 12755028 TI - [A case of subependymal giant cell astrocytoma not associated with tuberous sclerosis]. AB - The authors present a case of a subependymal giant cell astrocytoma (SGCA) not associated with tuberous sclerosis. On admission, a six-year-old boy had obstructive hydrocephalus caused by a huge intraventricular tumor. Preoperative T1-weighted MR images with gadolinium showed heterogeneous enhancement of the lesion. The tumor was totally removed through a right transcortical approach with frontal craniotomy. Immunohistochemical staining for glial fibrillary acidic protein (GFAP) showed a positive reaction in some of the large tumor cells and the labeling index of MIB-1 was < 1.0%. Histological examinations revealed SGCA. He has no other evidence of tuberous sclerosis at present, but the patient in this case is an infant, so there is a possibility of his being diagnosed as tuberous sclerosis in the future. Therefore, systemic regular follow-up is recommended, even when his condition is asymptomatic. PMID- 12755029 TI - [Solitary fibrous tumor of the fourth ventricle: case report]. AB - A rare case of solitary fibrous tumor, located wholly within the fourth ventricle, is reported. A 57-year-old male presented with headache and nausea. The preoperative magnetic resonance images revealed a well circumscribed mass in the fourth ventricle that exhibited a low intensity on T1-weighted images and homogeneously enhanced with gadolinium. Vertebral angiogram revealed a tumor stain supplied from the choroidal branches of the posterior inferior cerebellar artery. The tumor was totally resected through a midline suboccipital approach. Histologically, the tumor was composed of spindle-shaped cells growing in fascicles within a collagenous matrix. Immunohistochemical staining demonstrated vimentin and the CD34 positivity of tumor cells. Solitary fibrous tumor is a newly described entity, which should be considered in the differential diagnosis for dural-based lesions. PMID- 12755030 TI - [A successfully treated case of cerebrospinal fluid fistula caused by fracture of the sella turcica]. AB - Fracture of the sella turcica is rare and is associated with many complications. We successfully treated a cerebrospinal fluid fistula caused by a fracture of the sella turcica. A 66-year-old male in a motor vehicle accident was admitted to an outside hospital with disturbance of consciousness. A computed tomography (CT) scan of the head revealed a subarachnoid hemorrhage and pneumocephalus. Cerebrospinal fluid rhinorrhea developed after admission. Repair of the fistula was attempted without success, and the patient was transferred to our hospital for further examination and treatment. A fracture of the sella turcica was clearly visualized on coronal and sagittal head CT and on a three-dimensional reconstructed CT (3D-CT) image. The source of the CSF fistula was thought to be the anterior wall of the sella turcica. Through a bifrontal interhemispheric approach, the cerebrospinal fluid fistula was repaired microscopically with the assistance of endoscopy. Postoperatively, the fistula stopped completely. Coronal and sagittal head CT and 3D-CT images are useful for making a diagnosis of CSF leakage. Endoscopic images can assist in observation of the dead angle of the microscope. PMID- 12755031 TI - [Tumors (3), meningioma. no. 11 in series of articles: basic knowledge of neuropathology for neurosurgeons]. PMID- 12755032 TI - ["How I do it", no. 3: a case of cervical disc disease at multi cervical levels]. PMID- 12755033 TI - [Re-evaluation of clinical presentation in Spinocerebellar ataxia type 6]. PMID- 12755034 TI - [Correlation between inferior olivary hypertrophy and generation of palatal tremor (formerly termed 'palatal myoclonus')]. PMID- 12755035 TI - [The validation of a Japanese version of the Everyday Memory Checklist]. AB - We developed a Japanese version of the Everyday Memory Checklist(EMC), which was originally made by Wilson et al. to assess everyday memory problems, and examined its reliability and validity in assessing brain-damaged patients with memory deficits. SUBJECTS AND METHODS: The subjects consisted of 393 brain-damaged patients with memory deficits aged between 17 and 90 years with the mean MMSE of 24.6, and 132 control subjects aged between 19 and 86 years with the mean MMSE of 28.5. Patients' everyday memory problems were both self-evaluated and evaluated by their caregivers, while those of control subjects were only self-evaluated with the EMC. RESULTS: The test-retest reliability of the EMC evaluated in 149 randomly selected patients was acceptably high with intraclass correlation coefficient (0.950 for caregiver-evaluated scores and 0.759 for self-evaluated scores). The standard memory test scores were significantly correlated with the EMC scores of the patients evaluated by the caregivers; among them the scores of the Rivermead Behavioural Memory Test were most highly correlated with the EMC scores. On the other hand, the scores of the non-memory cognitive test scores were not significantly correlated with the EMC scores. The self-evaluated EMC scores of the patients were significantly lower than those of the patients evaluated by the caregivers, and were not correlated with memory test scores or with non-memory cognitive test scores. As the EMC scores of control subjects over 60 years were significantly higher than the scores of the subjects under 59 years, we compared the EMC scores between the patients and the control subjects after we divided each subject into two groups depending on age. The EMC scores of the patients evaluated by the caregivers were significantly higher than the score of the control subjects in groups of both over 60 years and under 59 years. However, the self-evaluated EMC scores of the patient were not significantly higher than the EMC scores of the control subjects. In the analysis of the diagnostic accuracy, the EMC scores of the patients evaluated by the caregivers correctly classified 40.9% of patients and 92.9% of normal subjects by setting the cut-off scores of 14/15 in subjects under 59 years, and classified 50.2% of patients and 85.5% of normal subjects by setting the cut-off scores of 16/17 in subjects over 60 years. CONCLUSION: When caregivers evaluated the amnesic patients, the Japanese version of the EMC is reliable and valid in assessing everyday memory problems. PMID- 12755036 TI - [Evaluation by statistical brain perfusion SPECT analysis on MRI findings, kana pick-out test and Mini-Mental State Examination results in patients with forgetfulness]. AB - The aim of this single photon emission computed tomography(SPECT) study was to determine the abnormality of the regional cerebral blood flow(rCBF) using a three dimensional stereotactic surface projection (3 D-SSP) in 18 patients who were referred to the hospital because of forgetfulness. Two intergroup comparison by 3 D-SSP analysis was conducted based on MRI, kana pick-out test and Mini-Mental State Examination (MMSE) results. Of the MRI findings, in the brain atrophy group, rCBF was decreased in the posterior cingulate gyrus, medial temporal structure and parieto-temporal association cortex; these rCBF-decreased areas are similar to the Alzheimer disease pattern. In the group where the MMSE was normal but the kana pick-out test was abnormal, rCBF was decreased in the posterior cingulate gyrus and cinguloparietal transitional area. In the group where both the MMSE and kana pick-out test were abnormal, rCBF was decreased in the parieto temporal association cortex, temporal cortex and medial temporal structure. These results suggest that 3 D-SSP analysis of the SPECT with MMSE and the kana pick out test provides the possibility of early diagnosis of initial stage of Alzheimer's disease. PMID- 12755037 TI - [Age-related working memory decline in patients with Parkinson's disease]. AB - The purpose of this study was to investigate the age-related working memory(WM) decline in patients with Parkinson's disease(PD) using Baddeley's WM model. This model consists of the central executive (CE) and two slave systems, the phonological loop (PL) for the storage of verbal materials, and the visuospatial sketchpad(VSSP) for the storage of visuospatial information. The participants of this study were 22 PD, 11 old (age of 68-78, mean age, 70.5) and age of onset, duration of illness, medication time, and Yahr stage, global cognitive status matched 11 young(age of 39-58, mean age, 51.5) PD, age- and educational years, global cognitive status-matched 22 normal control (NC), 11 old(age of 65-78, mean age, 70.4) and 11 young(age of 45-57, mean age, 52.4). Mental calculation span of digit sequences, digit forward and backward span, visual memory span were carried out. Age related decline of WM was found in both groups, but processing related differences were revealed between the two groups. NC group showed significant decline with aging in digit backward span. In contrast, in mental calculation span, PD groups showed significant deficit revealed in young PD group and declined significantly with aging and significant decline was not found in digit backward span. In term of the processing and difficulty of WM tasks, digit backward span that needs maintenance of digit sequences and re-ordering, was more difficult than mental calculation that needs maintenance of digit sequences, summation of the digit and updating of the results. There were not significant differences between four groups in digit forward span, visual memory span. The results indicated that the WM span in normal aging declined as task difficulty increased. Their performance decline may be caused by the CE dysfunction. On the other hand, PD showed a characteristic CE deficit observed in mental calculation even in young age and decline with aging. Such decline may be caused by peculiar processing related dysfunction of CE that assumes to be essential deficit of PD. PMID- 12755038 TI - [Unilateral multiple cranial nerve palsies mimicking Garcin syndrome as an atypical symptom of brainstem encephalitis: a case report]. AB - We report a 56-year-old Japanese man who was admitted because of dysphagia and left side facial dysesthesia. On admission, his general conditions were unremarkable. Neurological examination revealed that he was alert and well oriented. He exhibited left-side cranial nerve involvement such as the trigeminal(sensory and motor), glossopharyngeal and potential vagal nerve palsy. He exhibited neither long-tract signs, such as motor weakness, sensory disturbance in his exremities and pathological reflex, nor ataxia. By a few days after admission, his symptoms had worsened. Vertigo and diplopia appeared, and his consciousness level became drowsy. Bilateral third and sixth nerves, left fourth, fifth, eighth, ninth and tenth nerves were involved. Results of laboratory tests and CSF studies were within normal. Results of examination of the skull base X-ray and MRIs of the brain were normal. Administration of corticosteroid and intravenous administration of a high dose of gamma-globulin were not effective. His symptoms gradually recovered three months after admission. Based on clinical symptoms and results of physiological examination, i.e., an involvement of his consciousness and abnormal findings in blink reflex test that suggest involvement of the brainstem, he was diagnosed as having brainstem encephalitis. Although the exact pathophysiological mechanisms were unclear, it is clinically important to note that an atypical brainstem encephalitis may present a subacute progressive appearance of unilateral multiple cranial nerve palsies mimicking Garcin syndrome. PMID- 12755039 TI - [A case of the familial essential myoclonus and epilepsy presenting behavioral arrest]. AB - A 75-year-old Japanese man who had a past history of epilepsy was admitted because of rhythmical myoclonus of upper and lower extremities. His mother and son also had both epilepsy and myoclonus. On neurological examination, he was alert and oriented without dementia. There was no abnormal finding except for the myoclonic jerks appearing in his upper and lower extremities. The myoclonic jerks appeared at rest and worsened during maintaining posture. Results of laboratory tests were all within normal levels. Brain MRIs were normal, and his EEGs showed diffuse fast wave activities without paroxysmal discharges. Surface EMG studies revealed irregular myoclonic discharges at the frequency of 6-20 Hz. Electrophysiologically, he had a giant somatosensory-evoked potentials, an enhanced long-loop reflex(C-reflex) and cortical spikes preceding the myoclonic jerks, suggesting that his myoclonus was of cortical origin. Based on the cortical myoclonus, episodes of epilepsy and familiarity, he was diagnosed as having familial essential myoclonus and epilepsy(FEME). In addition, he periodically became a motionless state characterized by sudden appearance of transient akinesia with generalized stiffness of all of his limbs. The transient motionless state usually appeared following mental stress or sudden sensory stimuli such as loud sound. It persisted for about 10-20 min and ceased spontaneously. Since EEGs recorded during this akinetorigid state showed no paroxysmal discharges, it was considered not to be epileptic. However, after the administration of valproate, his cortical myoclonus improved, and the transient akinetorigid state also disappeared simultaneously. From symptomatic point of views, this generalized motionless state with stiffness of limbs may resemble to freeze behavior of wild animals suddenly confronted with sudden strong stimuli. From symptomatological as well as genetic aspects, FEME has been suggested to be heterogeneous. We propose that this unique akinetorigid attack might be a new clinical phenotype of FEME. PMID- 12755040 TI - [A Japanese family with familial Alzheimer's disease associated with presenilin 1 mutation: relationship between younger age of onset and ApoE gene polymorphism]. AB - We previously reported a Japanese family with early-onset familial Alzheimer's disease associated with G 209 R presenilin 1(PS 1) mutation. There have been six patients across three generations in this family. In the present report, we described the clinical course, findings with neuroimaging and results of genetic examination of PS 1 and apolipoprotein E(ApoE) in three of six patients(II-1, III 1 and 2). The clinical course was common to all three patients. Memory disturbance, disorientation, amnestic aphasia, personality changes and perseveration appeared at early stages, whereas Gerstmann's syndrome, myoclonus and general convulsion were recognized at advanced stages. CT disclosed mild brain atrophy in the temporal lobes at early stages and diffuse brain atrophy predominantly in the fronto--temporal lobes at advanced stages. SPECT exhibited hypoperfusion in the fronto-temporal areas at early stages and hypoperfusion in the fronto-temporal and parieto-occipital areas at advanced stages. The age of onset in six patients demonstrated two clusters at age 53-55(I-1, II-1, 2 and 5) and age 46-48(III-1 and 2). PS 1 genotyping demonstrated that the heterozygous exonic missense mutation G 209 R was confirmed in all three patients. Regarding the ApoE genotyping, II-1(mother) was epsilon 3/epsilon 3, whereas III-1 and 2(children) were epsilon 3/epsilon 4. These findings suggest the possibility that there might be a gene dose effect, since the age of onset ranged from 5 to 7 years younger in patients who received epsilon 4 alleles from the father. PMID- 12755041 TI - [A case of vertebral dissecting aneurysm manifesting as subarachnoid hemorrhage following nuchal pain]. AB - We report a case of subarachnoid hemorrhage (SAH) from vertebral dissecting aneurysm 4 days after first nuchal pain. The patient was a 46-year-old man with a sudden onset of nuchal pain. There were no obvious abnormalities detected on MR images in another hospital. Four days later, however, he was admitted to our hospital because of severe re-attack of nuchal pain. CT demonstrated moderate SAH and cerebral angiograms revealed right vertebral dissecting aneurysm. Proximal occlusion of the vertebral artery including its aneurysmal dilatation was performed using detachable coils. We strongly suspected that his initial symptom of nuchal pain was due to dissection of the vertebral artery itself, since the aneurysmal dilatation accompanied by intramural hematoma had been observed retrospectively in the initial MR imaging. The incidence of the vertebral dissecting aneurysm presenting with nuchal pain alone due to dissection is reported to be 7% in the literature. The prognosis of non-hemorrhagic vertebral dissecting aneurysm followed by delayed SAH is considered to be fatal. Therefore, careful investigations for differential diagnosis should be taken into account since the diagnostic possibility exists that non-hemorrhagic vertebral dissecting aneurysm would be manifested by a symptom of headache/nuchal pain alone. PMID- 12755042 TI - [A case of cranial fasciitis induced by trauma]. AB - We present a 20-month-old boy with a rapid growing occipital mass after head trauma, that was pathologically diagnosed as cranial fasciitis. Cranial fasciitis, first described as a specific entity by Lauer and Enzinger in 1980, is a benign fibroblastic lesion occurring in children that resembles nodular fasciitis pathologically. Nodular fasciitis also shows benign proliferation of fibroblasts and myofibroblasts in the subcutaneous tissues. This disease was firstly reported in 1955. This lesion is arising in the upper and lower extremities and trunk in adult commonly. The rapid growth, abundant cellularity, and mitotic activity cause these lesions to be misdiagnosed as sarcomas frequently. However, these lesions recur rarely, do not develop metastases, and are readily cured only by surgical excision. Cranial fasciitis is similar to nodular fasciitis in clinical cause and pathology. Therefore cranial fasciitis should be considered a variant of nodular fasciitis, although it differs from noduler fasciitis in its frequent site and age. Cranial fasciitis occurs in skull bone and in childhood. The cause of them are still unknown. However, we speculate on the relationship between head trauma and cranial fasciitis making reference to the fact that 5-15% of nodular fasciitis were caused by trauma. So we investigated this relationship in cranial fasciitis, then 8 cases of 40 cases(20%), that we could trace in the world literatures, had histories of previous head trauma. From this result, head trauma is one of a cause of cranial fasciitis probably. PMID- 12755043 TI - [A case of treatment on traumatic pseudoaneurysm of internal carotid artery occurred epistaxis]. AB - We reported a case of traumatic pseudoaneurysm of internal carotid artery accompanied by skull base fracture, for which Guglielmi detachable coil(GDC) embolization surgery was very effective for recovery. A 53-year-old man met a traffic accident and was admitted to our hospital with blindness and epistaxis accompanied with shock. Balloon occlusion test showed a positive sign of traumatic pseudoaneurysm of internal carotid artery, and under the diagnosis GDC embolization surgery was performed on six days after the onset. The patient has completely recovered and keeps his wellness untill now after six months from the surgery except for loss of left eye vision. We believe that the endovascular surgery using stents will become the prime modality for treatment of traumatic pseudoaneurysm in the future. PMID- 12755044 TI - [Acute facial paralysis as the initial presentation of parotid tumor]. PMID- 12755045 TI - [Arachnoid cyst associated with fibrous dysplasia]. PMID- 12755046 TI - [Relationship between doctors and patients]. PMID- 12755047 TI - [Clinical approach to improve quality of life of disabled children: introductory remarks]. AB - This symposium, discussed clinical approaches to improve the quality of life for severely disabled children including the management of feeding and breathing problems, school life and grief care. Pediatric neurologists should be health care professionals for them, consider improving their quality of life at both home and school, help them to decide which information from different professionals is appropriate. In order to improve their quality of life, feeding and breathing problems are often the most important. Furthermore, we have to promote discussion and co-operation with school teachers, especially about so called medical care. PMID- 12755048 TI - [Medical care and support in school and community life to very severe neurologically-impaired children--advance and problems in medical, educational and social management for improvement of QOL]. AB - With an increasing number of children with severe neurological impairment living in their houses, there is growing demand for medical care and support in school and community life. In such cases, respiratory disorder, gastro-esophageal reflux and dysphagia are closely related. To improve these disorders, appropriate rehabilitation and daily managements, such as posture control, are important as well as medical and surgical treatment. Social and educational support is also necessary for improvement of the QOL of these children and their family. For example, daily medical care such as tube feeding and sputum suctioning should be provided by school staffs. Pediatric neurologists should actively participate in such educational and social activities. PMID- 12755049 TI - [Management of nutrition in children and adults with severe motor and intellectual disabilities]. AB - In children and adults with severe motor and intellectual disabilities (SMID), management of nutrition is very important. We investigated the problem of long term use of tube feeding on which many of them depend because of swallowing dysfunction. Trace elements such as copper, zinc, selenium and long chain unsaturated fatty acid (omega-3 fatty acid) were often deficient. To evaluate their nutritional condition, we calculated the body fat mass by the method of Bioelectrical Impedance Analysis and measurement of subcutaneous fat thickness. Patients with the athetotic type of cerebral palsy had a lower level fat mass than those with the spastic type. Patients with a low body fat mass tended to require more energy than those with a high body fat mass. To improve the quality of life of persons with SMID, it is important to find clinical dysfunction related to the nutrition and to improve the nutritional condition immediately. PMID- 12755050 TI - [Gastroesophageal reflux in neurologically impaired children]. AB - We present here the indication, procedure and results of surgical treatment of gastroesophageal reflux (GER) for neurologically impaired children. We decide its indication based on clinical symptoms and findings of upper GI series, esophageal pH monitoring and GI fiberscopy, respecting the QOL of the patient and family. Laparoscopic fundoplication has become the first choice in surgical treatment of GER because of its good results. However, an anti-reflux procedure is not effective in patients with intractable aspiration. A laryngotracheal separation procedure should be applied for these cases. PMID- 12755051 TI - [Treatment of respiratory disturbance in children with severe physical disabilities to improve their quality of life]. AB - Children with severe physical disabilities frequently have respiratory problems which affect their quality of life (QOL). They commonly stem from central nervous system dysfunctions and/or severe motor disabilities, and consist of various impairments deriving primarily from central and motor dysfunctions, such as dysmyotonia, deformation, dysphagia, and gastro-esophageal reflux (GER), which often influence each other and result in respiratory insufficiency without adequate interventions. Aging is also an important factor to worsen respiratory involvements and to change their underlying pathophysiology gradually, even if the primary cause of the disability is non-progressive. A key to effective treatment and improvement of their QOL is to understand the pathophysiology. Evaluation is primarily based on regular physical examination. Other examinations include blood sampling to examine inflammation, nutritional state and blood gas analysis, round-the-clock SpO2 and TcPCO2 or EtCO2 monitoring, chest X ray and computed tomography, polysomnography, laryngoscopy, bronchoscopy, evaluation of swallowing function, and evaluation of GER. Postural control, relaxation, respiratory physiotherapy, and treatment of dysphagia are the most important and common therapeutic procedures. Treatment of GER, treatment of upper airway obstruction, oxygen therapy, tracheostomy, or mechanical ventilation may be needed. It is also important to take into account the possibility that the treatment procedures themselves could affect the patient's QOL. Counseling should be performed before and throughout the treatment process about how patient's and family's everyday life will be influenced positively and negatively by the treatment, and a multidisciplinary team should support all aspects of their needs. PMID- 12755052 TI - [Role of disabled children's school as regional special education center]. AB - The final report of the Collaborative Research Conference on Special Education in the 21th Century pointed out that supportive activities, such as education, welfare, medical care and labor should be strengthened to provide special support for disabled children, and that measures should be taken for school children requiring medical care in daily life, in response to an increasing number of children with severe and multiple disabilities and other social changes. Recent great progress in pediatric medicine has enabled these children to attend schools for the physically handicapped which provide more specialized educational support. At present, support for these children is still insufficient. This symposium discusses these problems as well as future prospects of disabled children's schools. PMID- 12755053 TI - [Terminal care for handicapped newborns and grief care for their families]. AB - Terminal care for the handicapped newborns and the grief care for their families have recently emerged as a medical problem. Questionaires to 13 parents who lost their children showed that they were unsatisfied with the care from doctors and nurses before and after the child's death. Medical approach to grief care includes changes in the quality of medicine for handicapped newborns. A system should be created to improve the relationship between the NICU pediatricians/nurses and the staffs engaged in early intervention prior to the newborns' death, which in turn strengthen the ties with a network supporting the families who have lost their newborns. PMID- 12755054 TI - [Studies on the adverse effects of fluvoxamine treatment in children with autistic disorder: correlation with genetic polymorphism in serotonin related genes]. AB - Selective serotonin re-uptake inhibitors (SSRIs) have recently been applied to the children with autistic disorder. To create better treatment, we studied here clinical adverse effects of fluvoxamine and correlated them with genetic polymorphism of two genes, the promoter region of serotonin transporter gene (5 HTTLPR) and serotonin 2A receptor gene (5-HT2AR). Twenty-eight subjects, consisting of 23 boys and 5 girls, aged from 3 to 18 years old diagnosed as having autistic disorder were analyzed during fluvoxamine administration. The dosages and duration of fluvoxamine treatment are 1.5 to 3 mg/kg/day and 2 weeks to 17 months (mean 7.9 months), respectively. There were several clinical adverse effects such as sleep disturbance in 9 cases, climb up to high places in 8, gastrointestinal symptoms in 6, hyperactivities in 5, excitement in 4, general fatigability in 2 and urticaria in 1. Medication was discontinued in 2 patients with fatigability and 1 with sleep disturbance, diarrhea and poor appetite. There was no significant correlation between genetic polymorphism in 5-HTTLPR and the occurrence of clinical adverse effects of fluvoxamine. However hyperactivity was significantly more frequent in the subjects with 102T/102T polymorphism of 5 HT2AR, and patients with sleep disturbance were significantly less frequent in the subjects with 102C/102C polymorphism. We conclude that the clinical adverse effects such as climb up to high places and hyperactivity during fluvoxamine treatment may be relatively specific in children, and that genetic polymorphism of 5-HT2AR may be related to the appearance of clinical adverse effects. PMID- 12755055 TI - [Rhythmic fast activity on EEG in a patient with Mollaret's meningitis]. AB - Mollaret's meningitis is a rare disease of unknown etiology, characterized by repeated aseptic meningitis with transient neurological symptoms and quick recovery. The patient, a 16-year-old boy, had episodes of acute encephalitis followed by complete recovery every year from the age of 13 to 16 years. The symptoms at onset were loss of consciousness, a generalized tonic clonic seizure and pyramidal signs. He was first admitted to our hospital at the age of 15 years. EEG in the early stage showed diffuse low-voltage rhythmic fast actvity. He recovered consciousness after a week. At the age of 16 years, he was admitted again because of high fever (39 degrees C). Soon after admission, he had generalized tonic-clonic seizure and was accompanied by alternating deep coma and delirium. Cerebrospinal fluid examination during early stage revealed mild leukocytic pleocytosis, elevation of protein and a high level of IL-6. although brain CT and MRI showed no abnormal findings. EEG showed intermittent, diffuse, middle-voltage, rhythmic fast activity. After 5 days, the neurological symptoms completely disappeared with EEG normalization. To our knowledge, this is the first report of rhythmic fast activity on EEG in Mollaret's meningitis. The clinical pictures of this case suggest a transient functional disorder of brainstem or cerebrum may occur in this condition. PMID- 12755056 TI - [Three cases of benign myoclonus of early infancy]. AB - Benign myoclonus of early infancy (BMEI) is a non-epileptic paroxysmal phenomenon. Some patients with BMEI were mistakenly treated as infantile spasms, because the fits resemble to tonic spasms in infantile spasms and they occur in cluster. However, the patients have normal development and no abnormal electroencephalograms (EEG), and the fits spontaneously subside without sequelae. There are only a few reports on BMEI, and it is not widely recognized in Japan. We report three cases of BMEI. All the cases were suspected to have infantile spasms from the characteristic features of paroxysmal events, and the parents had strong anxiety because of recurrent fits. However, the fits decreased dramatically in about three months, and spontaneously disappeared within one year without any sequelae. BMEI might be included in cases of suspected infantile spasms, and such patients should be followed by monthly EEG examinations and close observation for other seizure phenomena. To avoid unnecessary treatments, such patients should be observed without any therapeutic trials including antiepileptic drugs. PMID- 12755057 TI - [Photosensitivity in electroencephalogram of a child with 45, X/46, X, mar (X) Turner Syndrome]. AB - We report an 11-year-old girl with Turner syndrome with 45, X/46, X, mar (X) who had mental retardation. EEG showed remarkable provocation of paroxysmal activity by photic stimulation. Diffuse irregular poly-spike and wave bursts were elicited by photic stimuli, without accompanying by clinical seizure. PMID- 12755058 TI - [A boy with infantile-onset fibromuscular dysplasia showing recurrent cerebrovascular attacks]. AB - We report here a boy with infantile-onset fibromuscular dysplasia (FMD). At 8 months of age, he had the first cerebrovascular attack presenting with left facial palsy, followed by recurrent attacks of alternating hemiparesis. Involvement of systemic arteries was manifested by variable somatic symptoms: retarded growth of the right arm due to stenosis of the brachial artery, constipation and recurrent abdominal pain due to intestinal ischemia, and renovascular hypertension. Cerebral angiography disclosed stenosis of the bilateral internal carotid arteries and right vertebral artery, with development of moyamoya disease like collateral vessels. Systemic arteriography revealed stenosis of the right brachial and left femoral arteries, which was either tubular or mimicking a string of beads. This case was characterized by the early onset and involvement of many cerebral and systemic arteries resulting in severe and variable symptoms. PMID- 12755059 TI - [A five-year-old girl with epilepsy showing forced normalization due to zonisamide]. AB - A case of forced normalization in childhood is presented. When zonisamide was administered to a five-year-old girl with intractable epilepsy, disappearance of seizures was accompanied by severe psychotic episodes such as communication disturbance, personal relationship failure, and stereotyped behavior, which continued after the withdrawal of zonisamide. These symptoms gradually improved by administration of fluvoxamine, however epileptic attacks reappeared. Although most patients with forced normalization are adult and teenager, attention should be paid to this phenomenon as adverse psychotic effects of zonisamide even in young children. Fluvoxamine may be effective for the symptoms. PMID- 12755060 TI - [Four cases of Fukuyama type congenital muscular dystrophy with edema]. PMID- 12755061 TI - [A new anti-adenovirus drug: where is it?]. PMID- 12755062 TI - [Gene transfer into corneal endothelial cells by Helios gene gun]. AB - PURPOSE: To examine the efficiency of particle-mediated gene transfer into rabbit corneal endothelium with a Helios gene gun system. METHODS: Using gene gun, plasmid DNA coding for green fluorescent protein(GFP) was transferred into cultured rabbit endothelial cells and rabbit corneal grafts from their endothelial site. Then the transferred corneas were transplanted as autografts. The GFP expression was detected by fluorescence microscopy. Histological examination of corneas was performed by light microscope and scanning electron microscope. RESULTS: In cultured rabbit endothelial cells, the transfection efficiency without damage to cells was about 7% under optimized helium pressure of 120 psi. In ex vivo, the GFP expression was limited to endothelial cells of the grafts at 140 psi. The histological findings from light and scanning electron microscopy of the grafts showed no severe mechanical damage in endothelial cells penetrated by gold particles. CONCLUSIONS: The helios gene gun system successfully transferred genes into corneal endothelial cells. However, further studies will be required to obtain the stable results of the particle mediated gene transfer into the endothelial cells of corneal grafts for clinical practice. PMID- 12755063 TI - [Antiviral effect of sulfated sialyl lipid against a clinical strain of adenovirus]. AB - PURPOSE: Currently, there is no antiviral drug for adenovirus(AdV). We have reported that sulfated sialyl lipid(NMSO) 3, a NMSO, has an antiviral effect against AdV prototype strains. We evaluated the antiviral inhibitory effect and the mechanism of NMSO 3 against AdV strains from patients with conjunctivitis in vitro. METHODS: Viruses used for the experiment were clinically isolated AdV type 3(AdV 3), AdV type 4(AdV 4), type 8(ADV 8), AdV type 19(AdV 19), and type 37(AdV 37). We examined three antiviral agents, i.e., NMSO 3, cidofovir(HPMPC), and zalcitabine(ddC). 50% effective concentration(EC50), 50% cytotoxic concentration(CC50), and selectivity index(SI) of compounds were determined for AdV infection in HEp-2 cells using 3-(4,5-dimetyl-thiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) methods. We also evaluated the anti-AdV activity of NMSO 3 when it was added during the stage of virus adsorption. RESULTS: NMSO 3, HPMPC, and ddC showed an inhibitory effect against all five AdV clinical strains. The EC50 values of NMSO3 were lower than those of HPMPC and ddC. NMSO 3 exhibited minimal cytotoxicity. NMSO 3 inhibited AdV infection only when it was added during the stage of virus adsorption. CONCLUSIONS: NMSO 3 inhibited the replication of all clinical AdV serotypes tested. NMSO 3 was the most potent and selective anti-AdV compound. The mechanism of anti-AdV activity by NMSO 3 was inhibition of viral adsorption to cells. PMID- 12755064 TI - [Late onset diffuse lamellar keratitis]. AB - BACKGROUND: Diffuse lamellar keratitis (DLK) is marked by the presence of diffuse or multifocal infiltrates confined to the laser in situ keratomileusis(LASIK) interface. These infiltrates are culture-negative, and the etiology is thought to be noninfectious. Most cases of DLK occur within the first week or two following surgery. CASE: We describe two cases of diffuse lamellar keratitis that occurred 3 months after LASIK. These patients were treated with intensive topical corticosteroids. RESULTS: We treated the patients with topical corticosteroids, with rapid improvement in patient symptoms, visual acuity, and slit-lamp biomicroscopy. CONCLUSION: DLK may occur three months after LASIK. PMID- 12755065 TI - [A case of intractable recurrent corneal erosion caused by Munchausen syndrome]. AB - OBJECT: We report a case of intractable recurrent corneal erosion that was unresponsive to any medication, but was cured by psychiatric care after diagnosis of Munchausen syndrome. CASE: The patient, a 35-year-old female, who developed pain in her left eye in July, 20 XX, was diagnosed with epidemic keratoconjunctivitis at the hospital where she worked as a nurse. Despite medication, the eye condition gradually worsened. After undergoing examination at many medical institutions, she came to our hospital on October 6, 20 XX. Her left eye manifested superficial punctate keratitis, then exhibited recurring corneal erosion. The eye condition was intractable, total corneal epithelial defect ultimately occurring on December 30. Subsequently, a bottle of hydrochloric acid oxybuprocaine eyedrops was found under her pillow. We therefore suspected that the corneal erosion was due to self-inflicted injury. She consulted a psychiatrist, and was diagnosed with Munchausen syndrome. After about 2 months of therapy in which she was prevented from touching the eye(eye patch etc.), her eye condition improved. CONCLUSION: This case was not diagnosed as Munchausen syndrome for 5 months. In cases of intractable corneal erosion, the possibility of self-injury should be considered. PMID- 12755066 TI - [A case of steroid-induced glaucoma after radial keratotomy]. AB - BACKGROUND: We report a patient who was diagnosed as having steroid-induced glaucoma after radial keratotomy(RK) and suffered from severe visual field defect. CASE: A 29-year-old man underwent RK for both eyes. After the operation, he was treated for six months with topical medication including 0.1% and 0.01% betamethasone without measuring intraocular pressure(IOP). When he consulted an ophthalmologist, his IOP was 43 mmHg in the right eye and 51 mmHg in the left eye. At our initial examination, his IOP was 8 mmHg in the right eye and 10 mmHg in the left eye with 750 mg acetazolamide peroral, 0.5% timolol maleate, and latanoprost eyedrops. There were RK 16 incisions on the cornea and we found severe glaucomatous visual field loss. Finally we performed trabeculotomy in both eyes for IOP control with conservative therapy. CONCLUSION: As the keratorefractive surgery becomes popular, we must be careful of problems, such as steroid-induced glaucoma, and the change of refraction following the change of IOP. PMID- 12755067 TI - [Two cases of primary biliary cirrhosis accompanied with severe keratoconjunctivitis sicca due to Sjogren syndrome]. AB - BACKGROUND: Primary biliary cirrhosis(PBC) is occasionally associated with Sjogren syndrome and results in liver cirrhosis. It occurs particularly in women, middle-aged or older, and is characterized by the presence of anti-mitochondrial antibody (AMA). We diagnosed PBC in 2 patients with severe keratoconjunctivitis sicca (KCS). CASE 1: A 45-year-old woman was diagnosed with PBC. A test for the presence of AMA was positive and liver dysfunction was detected. Tests for the presence of anti-SSA antibody and anti-SSB antibody were also positive. Signs of severe sicca syndrome observed in the oral cavity and in the eyes were compatible with signs of Sjogren syndrome. Furthermore, superior limbic keratoconjunctivitis was also observed. CASE 2: A 57-year-old woman was diagnosed with PBC and Sjogren syndrome. She also had thyroiditis and severe KCS. Tests for the presence of AMA, anti-SSA antibody, and anti-SSB antibody were positive. In both cases, eye drops were not effective as a treatment for the KCS, but lacrimal punctal occlusion with cauterization was effective. CONCLUSION: PBC should be looked on as a disease that may possibly promote severe KCS. PMID- 12755068 TI - First report of a family with Lynch syndrome type II in Puerto Rico. AB - Colorectal cancer (CRC) incidence in Puerto Rico has increased prodigiously since incidence figures were first recorded in 1950. Implications for hereditary nonpolyposis colorectal cancer (HNPCC) in concert with this increased CRC incidence are discussed. A family with the Amsterdam-positive criteria of the Lynch syndrome II variant, identified in the eastern area of Puerto Rico, is described. As far as we can determine, this is the first such report of this disorder in Puerto Rico. PMID- 12755069 TI - Technetium-99m sestamibi in the diagnosis of breast cancer. The Mayaguez Medical Center experience. AB - The most successful screening procedures for breast cancer are breast physical examination and mammography. However, mammography has a positive predictive value of 15-30% for nonpalpable malignancy and of 22% for palpable carcinoma; this results in a large number of biopsies on patients with benign lesions. Furthermore, the benefit of mammography in woman with dense breast tissue (< 50 years old) has been questioned. Different studies have shown that Scintimammography with Tc-99m has a high sensitivity and specificity for detecting breast cancer (average of 85% and 89%, respectively), with higher positive predictive value for palpable lesions (89%). We reviewed retrospectively 35 records of patients that had Scintimammography in our institution, and the sensitivity, specificity, positive and negative predictive values were similar to other centers (100%, 76%, 62.5% and 100%, respectively). So, Scintimammography may be a complement for current diagnostic techniques for breast malignancy in our setting. PMID- 12755070 TI - Prevalence and correlates of DSM-IV substance use disorders in Puerto Rico. AB - We report the basic findings of a survey aimed at estimating rates of substance disorders in a probability sample of 4,709 household residents aged 15 to 64 years old. Lifetime use of alcohol was reported by 77.2%, and 10.7% reported ever using illicit drugs. Overall, 14.7% of the sample met criteria for a lifetime substance disorder, and 4.9% for a past year disorder. The rates of lifetime disorders were 13.1% for alcohol and 4.1% for illicit drug. Past year abuse/dependence was 4.3% for alcohol and 1.3% for illicit drugs. Alcohol use disorders were associated with male gender, higher family annual income, being employed, and being married. Illicit drug use disorders were associated with male gender and younger age. Only 13.0% of respondents with a past year disorder reported using services for their disorder. A program of continuous monitoring of substance using disorders is critical to establishing and monitoring effective policies. PMID- 12755071 TI - Presacral neuroblastoma in a child: a case report. AB - Presacral location of neuroblastoma is rare. Resection entails an abdomino-sacral approach. This case report discusses clinical, imaging and successful surgical management of a presacral neuroblastoma in a one-year-old male child. PMID- 12755072 TI - Carcinoid tumor in Puerto Rico; two case reports. PMID- 12755073 TI - ST. segment elevation: is it a possible infarct? AB - In patients with acute substernal pain seen at Emergency Departments, ST segment elevations are considered the hallmark of an acute myocardial infarct. Acute substernal pain associated with ST segment elevations is the inclusion criteria for thrombolytic therapy. However, there are other conditions, which may present with ST segment elevation in which thrombolytic therapy is not indicated. Acute pericarditis and ECG variants of normal must also be considered in the differential diagnosis. Three cases are presented that illustrate this ECG presentation. It is of paramount importance, that the Emergency Department physician who does the triage for these patients be able to identify the various causes of ST segment elevation. PMID- 12755074 TI - In utero demonstration of aberrant systemic blood supply in lung sequestration: a case report. AB - Prenatal diagnosis of a potentially lethal condition as shown and confirmed with color flow Doppler; demonstration of an aberrant systemic blood supply. PMID- 12755075 TI - [Drug therapy of chronic pain in the aged]. PMID- 12755076 TI - [The 50th Anniversary of S.E.R. (Sociedad de Educacion y Rehabilitacion). 10 September 2000]. PMID- 12755077 TI - Not all that wheezes is asthma. PMID- 12755078 TI - Becoming an RNABC workplace representative. PMID- 12755079 TI - The road to advocacy. PMID- 12755080 TI - Reporting HIV infections to be mandatory. PMID- 12755081 TI - Playing the system. PMID- 12755082 TI - From basic data to composite indices: a re-examination of mortality analysis. PMID- 12755083 TI - International Classification of Diseases: preparation of short lists for data tabulation. PMID- 12755084 TI - Development of the Healthy Condition Index using geographic information systems in health. PMID- 12755085 TI - Guidelines for surveillance, prevention and control of West Nile virus. PMID- 12755086 TI - Working toward peace in the clinical setting: the role of clinical ethics in conflict resolution. AB - Ethics consultants or committees are often called into situations of conflict. What is their role in conflict resolution? What process should they use? What standards should they apply? The methods of alternative dispute resolution (negotiation, mediation and arbitration) provide a useful model for analysis of procedure, though they may not adequately describe all ethics consultations. Boundaries of acceptable standards may be gleaned from the precepts of medical ethics as well as from statutory and case law. In addition, the believer may obtain guidance from Scripture and prayer. PMID- 12755087 TI - Dealing with death. PMID- 12755088 TI - Physician assisted suicide: a constitutional crisis resolved. PMID- 12755089 TI - From involuntary sterilization to genetic enhancement: the unsettled legacy of Buck v. Bell. PMID- 12755090 TI - Physician-assisted suicide legislation: issues and preliminary responses. PMID- 12755091 TI - Historical and Biblical references in physician-assisted suicide court opinions. PMID- 12755092 TI - Abortion, euthanasia, and the need to build a new 'culture of life'. PMID- 12755093 TI - Abortion, physician-assisted suicide and the Constitution: the view from without and within. PMID- 12755094 TI - Reading the mystery passage narrowly: a legal, ethical and practical argument against physician assisted suicide. PMID- 12755095 TI - IRBs and monitoring the safety of clinical trials. PMID- 12755096 TI - Research subject's complaint launches federal investigation. PMID- 12755097 TI - The Constant Gardener revisited: the effect of social blackmail on the marketing concept, innovation, and entrepreneurship. AB - This paper discusses how adoption of the social dimensions of the marketing concept may unintentionally restrict innovation and corporate entrepreneurship, ultimately reducing social welfare. The impact of social marketing on innovation and entrepreneurship is discussed using the case of multinational pharmaceutical firms that are under pressure when marketing HIV treatments in poor countries. The argument this paper supports is that social welfare may eventually be diminished if forced social responsibility is imposed. The case of providing subsidized AIDS medication to less developed nations is used to illustrate how social blackmail may result in less innovation, entrepreneurship, and product development efforts by the pharmaceutical industry, ultimately reducing social welfare. PMID- 12755098 TI - Reevaluating repugnance: a critical analysis of Leon Kass' writings on genetic reproductive technologies. AB - Recent philosophical and political discourse surrounding the issue of human cloning has dominated both public and academic arenas. With the dual potential to address human disease, distress, and disorder in unprecedented ways, and to offend a vast public uncomfortable with the technology, cloning represents a true political conundrum. Adulterated by the opinion of an uninformed public, the murky waters of public policy-making foster arguments like Leon Kass' "Wisdom of Repugnance." Unfortunately, such appeals to repulsion do not fare as well in the academic arena. Kass proposes that society yield to repugnance as an ethically relevant factor, since the elicitation of such repugnance signifies the defilement of human nature. However, such an application of repulsion and offense to human nature leads to improbable conclusions and internal contradictions that soundly repudiate the acceptance of such a principle. Thus, rather than pre analytically yielding to visceral emotions and passions, individual rational agents--and the public generally--ought to thoroughly analyze all of the relevant factors surrounding cloning before, and perhaps instead of, rejecting the technology simply because it elicits repugnance. PMID- 12755099 TI - [Postoperative measures to prevent recurrence of chronic pansinusitis and polyposis nasi]. PMID- 12755100 TI - Expression cloning of a novel suppressor of the Lec15 and Lec35 glycosylation mutations of Chinese hamster ovary cells. PMID- 12755101 TI - The ethics of cloning and human embryo research. AB - The successful cloning experiments that led to Dolly in 1997 have raised many ethical and policy questions. This paper will focus on cloning research in human embryonic cells. The possible gains of the research will be judged against the moral issues of doing research on a person. This paper concludes that while the embryo has some moral status, its moral status is outweighed by the multitude of benefits that embryonic stem cell research will bring to humanity. Policy suggestions are given for dealing with this new and developing field of stem cell research. PMID- 12755102 TI - Euthanasia: how proponents justify it and provide models for regulation. AB - Life prolonging advances in medicine have raised debate about the concept and clinical practice of euthanasia for terminally ill patients. This debate involves the need for protection of patients' rights in their delicate condition, while providing them with the right to end their own life, if they choose to do so. This paper addresses how proponents justify euthanasia in societies that have legalized it, while ensuring that patients asking for a merciful death do not get exploited by the system. Regulation models are thus adapted from the Dutch and Oregon medical systems. PMID- 12755103 TI - [Recurrent bloody nasal secretion]. PMID- 12755104 TI - Mental illness and the right to refuse lifesaving medical treatment. AB - The legal and ethical debate surrounding the right of mentally ill patients to refuse life saving medical treatment is one area in the spectrum of patient rights that the medical community has failed to fully explore. To better investigate this concept, it is important to first focus on the history of the right to refuse treatment for all patients. Case studies then explore arguments on both sides of the issue, and focus discussion on the inadequacies of the current standards, a need for further study and universal testing principles in order to provide all patients with the rights they deserve. PMID- 12755105 TI - [Submandibular tumor]. PMID- 12755106 TI - Biotechnology and the new politics of life and death in Brazil: the AIDS model. AB - The Brazilian government is providing free antiretroviral therapies to all of the country's registered AIDS cases, and this policy is being hailed as a model for treatment AIDS worldwide. My paper is an ethnographic investigation of this emergent biopolitical paradigm. I explore the forms of governance, civility, and humanness that emerge through this population whose rights are biotechnologically realized, and the mechanisms through which this model continues to not address the most vulnerable populations, leaving them "invisible." PMID- 12755107 TI - Cost cutting: substandard care. AB - Mr. Edwards is diagnosed with a heart arrhythmia and is treated with a successful regimen of medication. He joins an HMO and begins care under a new doctor who ponders whether he should switch Mr. Edwards' medical regimen. Changing the treatment would save the HMO money that could be allocated among other patients, but would expose Mr. Edwards to a small risk of fatal heart attack. I argue that the doctor should not change the medication without the patient's considerations for the following reasons: doctors ought to work solely for the welfare of the patient, the patient has no obligation to cut HMO costs; and allowing doctors' treatment decisions to be influenced by third a party would have negative social repercussions. Furthermore, an informative doctor-patient relationship would be preferred over other "relationship models" for determining Mr. Edwards' course of medication. PMID- 12755108 TI - Intention, manner, and culpability in death. AB - This paper criticizes the policy of the American Medical Association's policy on euthanasia. Contrary to a notable criticism of the AMA policy, this paper focuses not on the distinction between acts of commission and omission, but on the distinction between medical treatment aimed at ending life and medical treatment aimed at preserving an acceptable life for the patient. PMID- 12755109 TI - Reconstructing the stem cell debate. AB - Human embryonic stem cells have been a major topic in science, medicine, and religion since their discovery in 1998. However, due to the complex discourse and rhetoric of scientific language, debate has remained within the professional realm via "expert bioethics." Using the tenets of pragmatism, the author examines the need to move the debate to society as a whole and disentangle the stem cell debate from the ideologies of the human cloning and abortion debates. Opening this issue to a societal debate will advance societal growth, resulting in informed decisions on moral issues, funding, or regulation associated with hES cell research. PMID- 12755110 TI - [3-dimensional concept for abdominal ultrasonography. Consensus of the Section of Internal Medicine, Radiology and Surgery of the German Society of Ultrasonography]. PMID- 12755111 TI - Medical ethics and reproductive genetics in Swedish public discourse. PMID- 12755112 TI - How to talk about cloning without talking about cloning: public discourse in the UK. PMID- 12755113 TI - Budd-Chiari-Syndrome. PMID- 12755114 TI - Totipotent and differentiated cells: an ethical difference for therapeutic cloning? PMID- 12755115 TI - Bioethics or biopolitics? On the relationship between academic and public discussion of the "selection" and "manipulation" of human life. PMID- 12755116 TI - [The Ultrasonography Museum--a contribution to the development of diagnostic ultrasonography in Germany]. PMID- 12755117 TI - Bioethical issues in the relationship between Muslim patient and non-Muslim physician. PMID- 12755118 TI - Misreading the genetic blueprint: implications of genetics-based population screening. AB - Scheduled for completion in 2003, the Human Genome Project will provide unprecedented insight into the genetic basis of human disease. The choice of genomic medicine, however, must be explored, and not assumed to be the next logical step in the evolution of medicine. Genomic medicine epitomizes the dominant scientific paradigm of reductionism, the theory that the whole is best understood by breaking it down into its constituents. This concept makes genetics based disease prevention problematic as it tends to discount contextual and population determinants in pathogenesis--such as gender, class, and ethnicity--in favour of biological causes. Unqualified acceptance of a genetic guide to disease prevention is an ethical concern as it commits the "individualistic fallacy," and ignores contextual factors that may lead to genetic discrimination, and to the augmentation of social inequities. PMID- 12755119 TI - Pharmacogenetics and pharmacogenomics: recent developments, their clinical relevance and some ethical, social, and legal implications. AB - In recent debates on novel procedures of molecular medicine pharmacogenomics is attracting more and more attention as a genotype-based approach for improving safety and efficacy of the use of therapeutic substances. Promoted by basic knowledge generated in the field of medical genomics, facilitated by novel technological tools for mapping genetic variation in individuals, and supported by results of initial clinical studies linking specific genotypes to metabolic characteristics of individuals important for assessing drug response, procedures of pharmacogenetics and pharmacogenomics now are starting to impact significantly on clinical research and development and medical practice. In this situation assessing the goals, risk, and benefits of pharmacogenetics and pharmacogenomics is essential for the medically successful, ethically justifiable, and socially acceptable implementation of genotype-based diagnosis and pharmacotherapy. We discuss the current state of the art in pharmacogenetics and pharmacogenomics and introduce a model for evidence based assessment of its goals, risk, and benefits. We differentiate here between pragmatic and normative issues in the development of pharmacogenomics in order to contrast prevailing, insufficiently interest based modes of public technology assessment with the evidence-based mode that can be established as part of clinical study design. Finally, we provide a framework for the analysis of social accountability that can be used for technology development and technology assessment with regard to pharmacogenomics in particular and molecular medicine in general. PMID- 12755120 TI - Organ donation: a pilot study of knowledge among medical and other university students. AB - BACKGROUND: Physicians' knowledge of and attitudes towards organ donation may be a factor in organ procurement rates. There is a lack of information about how Canadian medical students perceive organ donation, and what they know about it. OBJECTIVE: This pilot study assesses the knowledge and attitudes of university students toward organ donation. METHODS: Medical students completed a self administered questionnaire after a lecture. Non-medical students completed the same questionnaire at the university student center. The questionnaire included a test that was used to assess knowledge about organ donation. Attitudes were assessed by determining whether the student carried a signed organ donor card, and their reasons if they did not. RESULTS: Of the 76 students in the first-year medical class, 39 responded. A sample of the first 40 non-medical students to visit a booth at the student centre was selected for comparative analysis. The mean age of medical students was 23.5 years; 23 for non-medical students. Of those surveyed, 56.6 per cent were women. Of medical students, 30.8 per cent reported carrying a signed card compared with 50 per cent of non-medical students. The most common reason for not carrying a card in both groups was apathy. Median test scores were 2.4/6 for both groups. Students carrying a signed card had a median test score of 2.7/6, with the median score for those not carrying signed cards being 2.2/6. CONCLUSION: More investigation of the knowledge and attitudes of medical students regarding organ donation is warranted. PMID- 12755121 TI - [The origin of cephalosporins]. AB - The origin of cephalosporins is investigated. In 1945, Giuseppe Brotzu, who was the rector of the University of Cagliari in Sardina, Italy, isolated a cephalosporin-producing strain, Cephalosporium acremonium. Although as many as 48 cephalosporin derivatives have been developed in Japan, how a cephalosporin producing organism was discovered is not widely known here. This article contains the first Japanese translation of Brotzu's Italian publication entitled "Ricerche su di un nuovo antibiotico (Research on a new Antibiotic)" and reveals how cephalosporin was developed, together with a cross reference to the first report of penicillin, a similar antibiotic compound, which was discovered by A. Fleming in 1928. Brotzu's brief academic and social background is also presented. PMID- 12755122 TI - Ethical behaviour of authors in biomedical journalism. AB - BACKGROUND: Biomedical journals communicate new information that changes health care decisions. If authors ignore the fundamental values of honesty and trust, that information becomes flawed, and society or patients may be harmed. OBJECTIVE: By describing two cases of unethical behaviour by authors, and using them as a focus to review acceptable ethics in publication, this article aims to educate readers who have not considered the ethical implications in writing manuscripts for biomedical journals. METHODS: Two cases of unethical behaviour by authors occurred when the results of new drug trials were reported. They were discovered after publication in a biomedical journal, and in the review process after the submission of a manuscript for publication respectively. In the first case, duplicate publication was identified because the same control data were used, but not acknowledged, in three publications by the same investigators. In the second, ghost writing by a pharmaceutical company writer was suspected because of the atypical presentation of a senior author's work. RESULTS: The editor consulted with the authors of both reports. In the first case, the authors concurred about the duplication, and the editors of the three journals wrote editorials to record the duplicate publications. The second case of ghost writing was unconfirmed by the authors, but the submission was withdrawn, and the article was later published in another journal. CONCLUSION: These cases draw attention to recently recognized types of scientific misconduct that influence the perception of scientific work. Duplicate publication and ghost writing not only deceive the reader, but may also conceal flawed study design and conflict of interest. PMID- 12755123 TI - [Camphor in the Edo era (4) moth repellent, deodorant, and fungicide]. AB - A troublesome task in the daily life of the Edo era was ridding houses of harmful insects such as mosquitoes, fleas and clothes moths. People commonly drove away mosquitoes by making smoke. They hung their clothes or books in the air to keep them free from moisture. This was effective in protecting them from becoming moldy or being damaged by insects. Various medicinal plants were used to eliminate harmful house insects or agricultural vermin. Camphor was a variety of insecticide, but it was not popular in early Edo times because it was not easily available then. But in the end of Edo period, camphor became popular as a moth repellent. PMID- 12755124 TI - A call to arms for drug discoverers. PMID- 12755125 TI - [Chen Zutian: the fourth heir to Chen Wailang]. AB - This manuscript deals with the state of affairs in the Chen-Wailang family, with special emphasis on Soden, the fourth heir to the founder of the Uiro family Chen Wailang. Especially interesting is the library that encompassed much of the medical literature of the time, which may still have a bearing on the modern world. PMID- 12755126 TI - Statins--the heart of the matter. PMID- 12755127 TI - Spirituality, religion, and health: a critical appraisal of the Larson reports. AB - The four-volume corpus The Faith Factor, and Scientific Research on Spirituality and Health: A Consensus Report by Larson et al constitute the largest English language review of research on spirituality and health. We have done a critique of the 329 systematic analyses of peer-reviewed research papers presented therein. The objectives were to determine if the Larson conclusions can be generalized; to document the understanding of the potential of qualitative research in assessing the spiritual domain; and to examine whether the definitions of religion and spirituality used by Larson et al correspond to those in general use. We conclude that their results cannot be generalized to other religious and cultural settings; that there is a need for more research focusing on age groups, cultures, religions, and clinical settings not adequately represented in studies to date; and that the need for more qualitative research methods justifies a detailed analysis of the use of qualitative methods in the studies reviewed by the Larson group. Finally, there is a need to establish a common vocabulary that bridges cultural and religious traditions, and facilitates clinical care, research, and teaching relating to spirituality, religion, and health. PMID- 12755128 TI - Enfuvirtide. PMID- 12755129 TI - Physician's ethical responsibilities when there is a discrepancy between demand and supply of medical services. AB - BACKGROUND: The discrepancy between the demand for and the supply of physician's services is the result of actions of multiple parties, including physicians. This situation raises ethical challenges for physicians, because it involves the profession's core values. OBJECTIVE: To discuss physicians' ethical obligations regarding the quantity of services that they might provide. METHOD: The obligations to accept new patients and to provide services to existing patients are deduced from the Code of Ethics of the Canadian Medical Association and The Royal College of Physicians and Surgeons of Canada. A model of physicians' ethics proposed by Pellegrino and Thomasma is examined as an example of a possible higher ethical standard. RESULTS: According to the Code of Ethics, physicians may refuse or accept new patients provided they do not discriminate improperly, and do not deny aid when there is "urgent need." It also states that the termination of medical services is permissible after "adequate notice" is given to the patient. The code states that physicians are to be guided by principles such as justice and compassion. Motivated by these principles, overworked physicians may accept new patients or delay the justifiable termination of services. Such action is praiseworthy as long as the physician can provide to all his or her patients the quantity and quality of care that meets professional standards. More altruistic ethical standards of practice than those in the Code of Ethics have been advocated. Physicians who, in making decisions about the quantity of services they provide, are guided by altruism rather than self-interest deserve recognition and encouragement. PMID- 12755130 TI - Does size really matter? PMID- 12755131 TI - Quality end-of-life care: the rights of every Canadian. PMID- 12755132 TI - Victorian naturalists in China: science and informal empire. AB - This paper discusses the research of British naturalists in China during the period between the Opium War and the collapse of the Qing dynasty (1839-1911). China was defeated in the Opium War and forced to open treaty ports for trade with the Westerners. The foreign powers, particularly Britain, imposed upon the Qing government treaties, concession leases, favourable trade conditions, legal privileges and so on to reduce its political autonomy. In the shadow of the informal empire, not only did the British have more freedom to travel in China, first at the treaty ports and later in the interior, but they successively established diplomatic , commercial and missionary institutions in dozens of Chinese cities. The most important of them - the British Consular Service, the Chinese Maritime Customs and the Protestant missionary organizations - provided the talent and infrastructure for natural historical research and became networks for scientific information. The research into China's natural history epitomized the characteristics of British research on China in general: it engaged in collecting and circulating an ever-increasing amount of information and aimed at producing 'factual' and 'useful' knowledge about China. The paper modified current literature on scientific imperialism, which has dealt primarily with the colonial context, by examining the role of nineteenth-century British imperial science in the context of informal empire. PMID- 12755133 TI - [Pancreatitis and pancreatic cancer]. PMID- 12755134 TI - NOD mice get the nod. PMID- 12755135 TI - [Definition of lipodystrophy syndrome in HIV-infected adults]. PMID- 12755136 TI - [Early detection of prostatic carcinoma: Is PSA screening suitable?]. PMID- 12755137 TI - [Prenatal care information: significance of the experimental health certificate of Seine-Saint-Denis (France)]. PMID- 12755138 TI - [Information systems Audipog as a tool for evaluation of the prenatal care network: experience of the Auvergne network]. PMID- 12755139 TI - Recent references. PMID- 12755140 TI - Importance of LDL/HDL cholesterol ratio as a predictor for coronary heart disease events in patients with heterozygous familial hypercholesterolaemia: a 15-year follow-up (1987-2002). AB - This study evaluated the prognostic significance of several risk factors on the outcome of coronary heart disease (CHD) in 639 cardiovascular disease-free subjects with heterozygous familial hypercholesterolaemia (FH). During the 15 year follow-up, 53 (18%) men and 34 (9.8%) women had a CHD event (men vs women, p < 0.001). The age-adjusted 15-year event rate was 3% (87 events/2915 person years). Smoking increased the CHD risk (hazard ratio = 2.45, p < 0.001), women had a 74% lower risk of a vascular event, compared to men, after controlling for the postmenopausal status (hazard ratio = 0.26, p < 0.001). A one-unit difference in low density lipoprotein (LDL)/high density lipoprotein cholesterol (HDL) cholesterol ratio was associated with a 17% higher risk (hazard ratio = 1.17, p < 0.05); hypertension increased the risk for an adverse event (hazard ratio = 3.02, p < 0.05) and a 1 mg/dl increase in plasma fibrinogen level was associated with a 4% higher CHD risk (hazard ratio = 1.04, p < 0.05). With the power of the 15 years of prospective evaluation, the study shows that increased smoking, hypertension and LDL cholesterol levels eight times more than HDL cholesterol predicts an adverse CHD event, in patients with FH. PMID- 12755141 TI - Cultural pluralism in health care: a South-African-Canadian comparison. AB - Views on the acceptance of cultural pluralism in health care are shaped by myriad of social and cultural factors. Through the comparison of Canada and South Africa, this article examines how ideology, history, demographics, and the cultural understanding of illness have shaped the views of cultural pluralism in South Africa in a way that is distinct from the Canadian perspective. Canadian health-care workers must consider such differences as we must be careful not to apply the concept of cultural pluralism in a way that people of other cultures may not understand or value. As the breadth of Canadian international health-care initiative grows, it is important to consider how other nations perceive cultural pluralism; otherwise, the application of our views on pluralism in health care can fail. PMID- 12755142 TI - The genome and the law: should increased genetic knowledge change the law? PMID- 12755144 TI - Assisted reproductive technologies in Nigeria: placing the law above medical technology. PMID- 12755145 TI - Unemployable genes: genetic discrimination in the workplace. PMID- 12755146 TI - "First, do no harm"--the fiction of legal parental consent to genital-normalizing surgery on intersexed infants. PMID- 12755147 TI - Testing pregnant women and newborns for HIV: legal and ethical responses to public health efforts to prevent pediatric AIDS. PMID- 12755148 TI - Are pitfalls of oxcillometric blood pressure measurements preventable in children? PMID- 12755149 TI - Corralling Kevorkian: regulating physician-assisted suicide in America. PMID- 12755150 TI - Recent advances in breast cancer: the 27th ESMO Congress 2002. AB - Breast cancer was the focus of several presentations during the 27th European Society of Medical Oncology (ESMO) Congress held in Nice. Fulvestrant seems to be equally effective to tamoxifen as a first-line therapy in hormone-sensitive advanced breast cancer in postmenopausal women. However, it may be more effective than tamoxifen in ER- and PgR-positive disease. The addition of tamoxifen to epirubicin containing regimens was found to significantly increase PFS and OS in patients with MBC according to a multivariate analysis of 640 patients. BRCA-1 linked breast cancer was reported to be more sensitive to anthracycline-based CT than sporadic and BRCA-2 linked disease. VEGF was found to be an independent prognostic factor in patients with node-negative breast cancer. The implications and limitations of these findings are discussed. PMID- 12755151 TI - Shaken baby syndrome: a comparative study: Anglo-American law and Jewish law- legal, moral, and ethical issues. PMID- 12755152 TI - Regulating advanced reproductive technologies: a comparative analysis of Jewish and American law. PMID- 12755153 TI - Opportunity lost: why and how to improve the HHS-proposed legislation governing law enforcement access to medical records. PMID- 12755154 TI - Human embryonic stem cell therapy. AB - The author presents an overview (completed on September 15, 2001) of three issues involved in the ethics of human embryonic stem cell therapy: the ethical implications of some of the scientific issues involved, the specific ethical issues of the moral standing of the early human embryo and the problem of cooperation, and a consideration of two public policy issues: should the research go forward, and what kind of health care system should the United States adopt. The author argues that the public policy questions are the most important agenda. PMID- 12755155 TI - Recent advances in breast cancer: the 25th Annual San Antonio Breast Cancer Symposium. AB - Some of the important developments in breast cancer treatment and biology are highlighted from presentations made at the 25th San Antonio Breast Cancer Symposium 2002 (San Antonio, Texas, 11-14 December 2002). These include an updated analysis of the ATAC trial, new data demonstrating the superiority of dose-dense adjuvant chemotherapy over conventional scheduling, and recent developments in the field of proteomics. PMID- 12755156 TI - Growing pains: disputes surrounding human reproductive interests stretch the boundaries of traditional legal concepts. PMID- 12755157 TI - Good-bye Dolly. PMID- 12755158 TI - Spontaneous portal-venous shunt of the liver. PMID- 12755159 TI - Hand fracture management. PMID- 12755160 TI - Fracture healing: bone healing, fracture management, and current concepts related to the hand. AB - Bones fracture frequently and often result in significant impairments, functional limitations, and disabilities, especially when the hand is involved. When fractures occur, there is a disruption of the skeletal tissue organization and a loss of mechanical integrity. The goal of fracture healing is to regenerate mineralized tissue in the fracture area and restore mechanical strength to the bone. Of equal importance is the reconstitution of the normal soft tissue gliding and movement about the fracture site. This article briefly reviews the history of fracture healing and the advances in mechanics and cellular and molecular biology, which should help the reader better understand the current mechanisms related to bone healing (primarily and secondarily). Fracture fixation modes also are described along with the temporal sequencing as to when to protect or move the fractured region. PMID- 12755161 TI - Principles of internal fixation as applied to the hand and wrist. PMID- 12755162 TI - Rehabilitation approaches for distal and middle phalanx fractures of the hand. AB - This article addresses current approaches for the rehabilitation of distal phalanx and middle phalanx fractures, excluding proximal interphalangeal joint avulsion fractures. Emphasis is placed on establishing the optimal rehabilitation program based on an understanding of the type and location of the fracture, method of fracture management, fracture stability, fracture healing, complications from soft tissue injuries, and recognizing and preventing common problems associated with fractures. Patient case examples and results reported in the literature are included. PMID- 12755163 TI - Management of proximal interphalangeal joint fractures and dislocations. AB - Proximal interphalangeal joint (PIP) injuries are among the most common in the hand and their severity is often underestimated. These injuries often lead to prolonged disability, pain, and stiffness. Appropriate treatment includes a thorough assessment, physical examination, and directed imaging. Such an approach should lead to a rational treatment plan that focuses on the rehabilitation of all damaged components, including osseous, articular, and soft tissue structures. This article reviews all elements in the management of PIP injuries and introduces an assessment method for PIP injuries based on the mechanism of injury rather than primarily on the basis of radiographic findings. PMID- 12755164 TI - Rehabilitation for proximal phalangeal fractures. AB - Proximal phalangeal fracture stability is crucial for the initiation of early and effective exercises designed to recover digital and especially proximal interphalangeal joint motion. Active digital flexion and extension exercises are implemented by synergistic wrist motion. Joint blocking exercises and active tendon gliding exercises in protective blocking splints are instrumental elements of early treatment. Dynamic splinting and serial finger casting are used in recalcitrant, severe, and late presenting cases. Surgical release is a last resort in regaining proximal interphalangeal joint motion. This measure is reserved for a failure of treatment when residual proximal interphalangeal joint contracture is persistent and severe enough to cause serious impairment of digital motion and hand function. PMID- 12755165 TI - Management of metacarpal fractures. AB - Fractures of the hand are the most common fractures of the human skeleton. Metacarpal fractures account for 30% to 50% of all of hand fractures. The mechanisms of these injuries vary from axial loading forces to direct blows to the dorsal hand. Resulting deformities include malrotation, angulation, and shortening. Treatment modalities vary from nonoperative reduction to open reduction and internal fixation. The treatment algorithm is guided by the location of the fracture, the stability of the fracture, and the resultant deformity. Operative procedures, although they may lead to excellent radiographic reduction of fractures, often lead to debilitating stiffness from the inflammatory reaction of the surgical procedure. Operative fixation must be employed judiciously and offered only when confident that non-operative therapy can be improved on with operative intervention. This article reviews the various types of metacarpal fractures, with the treatment options available for each fracture. The indications for each treatment modality, postoperative care, and rehabilitation are presented. PMID- 12755166 TI - An update on the management of carpal fractures. AB - This article reviews the appropriate management of common carpal fractures. The fundamental principles used by clinicians to choose appropriate stabilization and length of immobilization for these various injuries are discussed. The principles behind the progression of therapeutic intervention and the relation of progression to the stages of healing are emphasized. PMID- 12755167 TI - Early controlled mobilization of potentially unstable extra-articular hand fractures. AB - This article presents early controlled mobilization options for potentially unstable, nondisplaced, nonarticular hand fractures. Early controlled mobilization of tissues surrounding a healing fracture has the potential to enhance the quality and rate of fracture healing and a person's functional recovery. The options discussed protect the integrity of the fracture alignment, while permitting safe, pain-free protected motion of joints adjacent to the fracture. Traditionally, healing fractures are thought of as clinically stable or unstable. If clinically unstable, the fracture often is considered unable to tolerate unrestricted active motion during the initial stages of healing. This article offers an alternative perspective, in which clinicians can consider the clinical factors that can be controlled to allow for early protected motion of the regional tissues surrounding a potentially unstable hand fracture. These additional clinical options offer an alternative to acute fracture immobilization and help progress the rehabilitation of hand fracture patients. PMID- 12755168 TI - Metacarpal fractures in athletes: treatment, rehabilitation, and safe early return to play. AB - Specialty gloves and buddy taping of adjacent fingers may cushion impact and dissipate twisting forces so that hands, thumbs, and fingers are protected during play. When fractures occur, athletes must be protected from contact until healing has progressed to a point where reinjury or complications are unlikely and performance standards and expectations can be met. This article outlines a program of fracture management principles and progressive graduated rehabilitation that phase the hand-injured athlete first into general conditioning and non-ball-handling drills, then into return to hand impact activities, body contact, ball handling, and catching with the use of protective hand gear. At this point, specialized playing orthoses, gloves, or taping may be added to the treatment regimen. Batting, the use of golf clubs, and racquet handling occur later, and finally throwing with precision, distance, velocity, and frequency is initiated. The importance of the particular sport, the position played, and hand dominance are factored into the decision-making processes. Safety of the patient and opponents is paramount. Extra-articular metacarpal fractures are discussed as a prototype for treatment, rehabilitation, and early safe return to play because of their frequent occurrence in athletes. PMID- 12755169 TI - Bone graft substitutes. AB - A better understanding of the biology of fracture healing and an increasing awareness of the limitations and potential complications of autogenous bone graft harvest have combined to foster a burgeoning interest in the development of bone graft substitutes. A few of these materials have been available for more than a decade, and many more should become available in the near future. The characteristics of the ideal bone graft substitute may vary considerably depending on the intended site of application and the clinical setting in which it is used. Knowledge of the available alternatives is a necessary prerequisite to informed decision making. PMID- 12755170 TI - Complete nucleotide sequence of the chloroplast genome from a leptosporangiate fern, Adiantum capillus-veneris L. AB - We determined the complete nucleotide sequence of the chloroplast genome of the leptosporangiate fern, Adiantum capillus-veneris L. (Pteridaceae). The circular genome is 150,568 bp, with a large single-copy region (LSC) of 82,282 bp, a small single copy region (SSC) of 21,392 bp and inverted repeats (IR) of 23,447 bp each. We compared the sequence to other published chloroplast genomes to infer the location of putative genes. When the IR is considered only once, we assigned 118 genes, of which 85 encode proteins, 29 encode tRNAs and 4 encode rRNAs. Four protein-coding genes, all four rRNA genes and six tRNA genes occur in the IR. Most (57) putative protein-coding genes appear to start with an ATG codon, but we also detected five other possible start codons, some of which suggest tRNA editing. We also found 26 apparent stop codons in 18 putative genes, also suggestive of RNA editing. We found all but one of the tRNA genes necessary to encode the complete repertoire required for translation. The missing trnK gene appears to have been disrupted by a large inversion, relative to other published chloroplast genomes. We detected several structural rearrangements that may provide useful information for phylogenetic studies. PMID- 12755171 TI - Complete sequence and analysis of the plastid genome of the unicellular red alga Cyanidioschyzon merolae. AB - The complete nucleotide sequence of the plastid genome of the unicellular primitive red alga Cyanidioschyzon merolae 10D (Cyanidiophyceae) was determined. The genome is a circular DNA composed of 149,987 bp with no inverted repeats. The G + C content of this plastid genome is 37.6%. The C. merolae plastid genome contains 243 genes, which are distributed on both strands and consist of 36 RNA genes (3 rRNAs, 31 tRNAs, tmRNA, and a ribonuclease P RNA component) and 207 protein genes, including unidentified open reading frames. The striking feature of this genome is the high degree of gene compaction; it has very short intergenic distances (approximately 40% of the protein genes were overlapped) and no genes have introns. This genome encodes several genes that are rarely found in other plastid genomes. A gene encoding a subunit of sulfate transporter (cysW) is the first to be identified in a plastid genome. The cysT and cysW genes are located in the C. merolae plastid genome in series, and they probably function together with other nuclear-encoded components of the sulfate transport system. Our phylogenetic results suggest that the Cyanidiophyceae, including C. merolae, are a basal clade within the red lineage plastids. PMID- 12755172 TI - Peas-Mea1-Ppp2r5d overlapping gene complex: a transposon mediated-gene formation in mammals. AB - Human and mouse MEA1/Mea1 is flanked by two overlapping genes, a novel PEAS/Peas in a head-to-head orientation and PPP2R5D/Ppp2r5d in a tail-to-tail orientation making a Peas-Mea1-Ppp2r5d overlapping gene complex (PMP-complex). Genomic zoo blot analyses and database searching revealed that Mea1 exists only in mammals, while Peas and Ppp2r5d are conserved in eukaryotes. Mea1 and Peas are transcribed from a testis-expressed bidirectional promoter. Mea1-Ppp2r5d overlapping segment (MPOS) contains polyadenylation signals for both genes and shows marked conservation throughout mammals. Furthermore, the MPOS occupies 3'-region of transcripts of both genes is expected to form a clover-like intramolecular structure. Mouse genomic library Screening and database searches identified two MPOS-derived sequences in Odf2 gene and RP23-86H7 cosmid clone, respectively, in which MPOS might be a core segment for the retropositions. Thus, a key role of MPOS, a short transposable element containing polyadenylation signals on both strands, in the formation of the Mea1 during mammalian evolution is suggested. PMID- 12755173 TI - Alternative brain organization after prenatal cerebral injury: convergent fMRI and cognitive data. AB - The current study presents both longitudinal behavioral data and functional activation data documenting the effects of early focal brain injury on the development of spatial analytic processing in two children, one with prenatal left hemisphere (LH) injury and one with right hemisphere (RH) injury. A substantial body of evidence has shown that adults and children with early, lateralized brain injury show evidence of spatial analytic deficits. LH injury compromises the ability to encode the parts of a spatial pattern, while RH injury impairs pattern integration. The two children described in this report show patterns of deficit consistent with the site of their injury. In the current study, their longitudinal behavioral data spanning the age range from preschool to adolescence are presented in conjunction with data from a functional magnetic resonance imaging (fMRI) study of spatial processing. The activation results provide evidence that alternative profiles of neural organization can arise following early focal brain injury, and document where in the brain spatial functions are carried out when regions that normally mediate them are damaged. In addition, the coupling of the activation with the behavioral data allows us to go beyond the simple mapping of functional sites, to ask questions about how those sites may have come to mediate the spatial functions. PMID- 12755174 TI - Facial affect recognition deficits in bipolar disorder. AB - Patients diagnosed with bipolar disorder (BPD), by definition, have problems with emotional regulation. However, it remains uncertain whether these patients are also deficient at processing other people's emotions, particularly while manic. The present study examined the ability of 25 manic bipolar patients and 25 healthy participants on tasks of facial recognition and facial affect recognition at three different presentation durations: 500 ms, 750 ms, and 1000 ms. The groups did not differ in terms of age, education, sex, ethnicity, or estimated IQ. The groups did not differ significantly on either a novel computerized facial recognition task or the Benton Facial Recognition Test. In contrast, the bipolar group performed significantly more poorly than did the comparison group on a novel facial affect labeling task. Although the patient group had slower reaction times on all 3 computerized tasks, the presentation duration did not have an effect on performance in the patients. This study suggests that patients with bipolar disorder are able to recognize faces, but have difficulty processing facial affective cues. PMID- 12755175 TI - The hand movement test as a tool in neuropsychological assessment: interpretation within a working memory theoretical framework. AB - While the Kaufman Hand Movements Test (KHMT) is argued to be sensitive to cognitive impairment secondary to various forms of brain dysfunction in adults, it is unknown which cognitive processes it addresses. Dual-task research has employed tasks similar to the KHMT to determine whether such tasks assess (1) verbal or visuospatial memory, or (2) a proposed additional short-term memory component, movement memory. However findings consistent with both hypotheses have been reported. Experiment 1 involved 24 participants completing a hand movement task and a letter span task under articulatory suppression, finger movement and no interference conditions. Performance on both the hand movement task and letter span task was significantly reduced by articulatory suppression. In Experiment 2, 16 participants were administered the hand movement task and a Corsi span task under articulatory suppression, finger movement, spatial tapping, and no interference. Again, hand movement span was most reduced by articulatory suppression, in contrast to Corsi span which was most reduced by spatial tapping. Hand movement task performance was therefore assumed to rely upon verbal recoding strategies and thus the proposition of an additional component of movement memory was not supported. PMID- 12755176 TI - Motor adaptation in children with myelomeningocele: comparison to children with ADHD and healthy siblings. AB - Myelomeningocele is a common developmental malformation of the central nervous system that usually results in motor deficits. Previous studies of myelomeningocele have not examined motor adaptation, which involves changes in the control of movements that occur as a result of repeated task exposure but do not depend on conscious recall of the exposure. We studied motor adaptation in 17 children with myelomeningocele and shunted hydrocephalus, 19 children with attention deficit/hyperactivity disorder (ADHD), and 20 healthy siblings. All children were 8 to 15 years of age. They were administered 2 measures of motor adaptation known to be sensitive to subcortical abnormalities in adult neurological disorders. One task assessed the biasing in weight judgments that occurs after exposure to heavy versus light weights, and the other assessed the adaptation in reaching movements that occurs when vision is laterally displaced by prisms. Contrary to expectations, the groups did not differ in motor adaptation. Children in all 3 groups displayed significant biasing in their weight judgments and improvement in the accuracy of pointing during prism adaptation trials. Performance on the 2 motor adaptation tasks was not related to age or IQ. Weight biasing was positively related to a measure of response disinhibition. The findings suggest that myelomeningocele does not result in global impairment of motor skills, but instead in a profile of intact and impaired motor functions that potentially may be decomposed in accordance with the neuroscience of motor skills. PMID- 12755177 TI - Do children with myelomeningocele and hydrocephalus display nonverbal learning disabilities? An empirical approach to classification. AB - As a group, children with myelomeningocele (MM) and early-onset hydrocephalus demonstrate many of the features of the syndrome of nonverbal learning disabilities (NLD). However, the frequency with which individual children display a pattern of neuropsychological functioning consistent with the NLD syndrome is unknown. We addressed this question by comparing the prevalence of NLD in 32 children with MM and shunted hydrocephalus to that in a group of 27 healthy siblings. Participants, who were between 8 and 15 years of age, completed a neuropsychological test battery that included 11 measures of possible assets and 17 measures of possible deficits that define the NLD syndrome. As a group, children with MM and shunted hydrocephalus displayed many of the specific assets and deficits. However, they also displayed significantly more variability in their patterns of assets and deficits than siblings, reflecting the substantial individual differences that characterize children with MM. About 50% of the children with myelomeningocele displayed a pattern of assets and deficits consistent with the NLD syndrome. Classification as NLD was weakly related to cumulative medical risk, as well as to left-handedness. The findings suggest a need for caution in making generalizations regarding the applicability of the NLD model to children with MM and early-onset hydrocephalus. PMID- 12755178 TI - Coexistence of posttraumatic stress disorder and traumatic brain injury: towards a resolution of the paradox. AB - The coexistence of posttraumatic stress disorder (PTSD) and traumatic head or brain injury (TBI) in the same individual has been proposed to be paradoxical. It has been argued that individuals who sustain a TBI and have no conscious memory of their trauma will not experience fear, helplessness and horror during the trauma, nor will they develop reexperiencing symptoms or establish the negative associations that underlie avoidance symptoms. However, single case reports and incidence studies suggest that PTSD can be diagnosed following TBI. We highlight critical issues in assessment, definitions, and research methods, and propose two possible resolutions of the paradox. One resolution focuses on ambiguity in the criteria for diagnosing PTSD. The other involves accepting that TBI patients do experience similar symptoms to other PTSD patients, but that there are crucial differences in symptom content. PMID- 12755179 TI - Military and disaster nursing. PMID- 12755180 TI - September 11th--the Pentagon disaster: response and lessons learned. AB - September 11, 2001 is a date we will all remember. For the nurses at the Virginia Hospital Center-Arlington, however, the day is particularly important, as these nurses work at the official hospital for the Pentagon. This article provides an insider's view of the events of the day and the lessons learned. PMID- 12755181 TI - Preparing to respond: Joint Trauma Training Center and USAF Nursing Warskills Simulation Laboratory. AB - Injuries related to the events of September 11, 2001, and continuing military actions associated with Operation Enduring Freedom underscore the accurate focus of the Joint Trauma Training Center and the Warskills Simulation Laboratory. These two programs ensure that nurses are prepared to respond to diverse medical situations worldwide. Outcome measures from both initiatives attest to the effectiveness of an integrated program that facilitates critical thinking skills and clinical judgment to increase the nurses' ability to provide trauma care to severely injured military personnel. PMID- 12755182 TI - The relationship between proinflammatory mediators and heat stress induced rhabdomyolysis in exercising marines. AB - The problem of heat stress continues to plague military combat units deployed worldwide. Heat stroke is the most severe form of heat illness and carries 10% mortality despite intervention. Proteolytic and pyrogenic pro-inflammatory mediators, such as IL-6, are released into peripheral circulation in heat illness. It is not known how these mediators affect the outcome of heat illness. Identifying relationships between release of proinflammatory mediators and extent of rhabdomyolysis may lead to an intervention to reduce severity of outcome. This article presents the most current knowledge regarding the physiologic and pathophysiologic manifestations of heat illness. Based on this current state of the science the article introduces a study that will be conducted to address military unique heat stress research. PMID- 12755183 TI - Care of the critically ill patient in a military unique environment: a program of research. AB - The goal of the Air Force Nursing Research Program at WHMC is to conduct research on topics unique to Air Force and military nursing. The nine stressors of flight and the military environment of care have been used as a conceptual model to guide the development of research studies. The studies conducted to date describe how the environment affects practice and when the environment directly affects the patient. The studies conducted are examples of the numerous military nursing research projects supported by funding from the TSNRP. The research funded by TSNRP contributes to the body of nursing knowledge by supporting scientific research, particularly knowledge that is unique to the military. As our nation faces the threat of chemical and biologic attacks, terrorism, and increased deployment of soldiers to battlefields in remote locations throughout the world, it is more important than ever that we ensure the advancement of military nursing research. Supporting research that advances healthcare in peace and in war is critical to the care of our military members and their families. This will require that research funds continue to be available to support military nursing research, that a strong infrastructure to provide resources in support of nursing research programs continues to exist, and that the military nursing corps continues to attract, train, and retain PhD prepared nurse researchers. Given the results of the research completed to date, the following evidence-based practice can be applied to the care of the patient described at the beginning of this article: The nurse positions the patient in the center of the cargo compartment, away from the bulkhead, toward the front of the aircraft, the warmest location during flight. While enroute, the patient will need to be positioned on an aerovac mattress, repositioned frequently, and have his/her heels elevated at all times. Additional padding may be needed for areas adjacent to the litter cross members to reduce pressure on the skin in areas prone to pressure ulcer formation. Should the patient need endotracheal suctioning, the nurse knows that hyperoxygenation-hyperinflation is effective in preventing suctioning-induced hypoxemia. In addition, the suction pressure will need to be increased to account for the effects of altitude without exceeding the pressure limits on the transport ventilator and causing catastrophic ventilator failure. Because there is not enough room on the litter for the chest tube drainage tubing to lay straight, it will be coiled and should dependent loops develop, they should be drained every 15 minutes. This is Air Force nursing research in practice. PMID- 12755184 TI - Managing critical care casualties on the Navy's hospital ships. AB - In this article, a history of the hospital ships was recounted. Recent missions were described in terms of the ship and crew's capability based on education and mock training exercises. Patient flow was described and a case scenario was presented to illustrate surgically intensive management of critical care casualties. Finally, thoughts on the future of hospital ships were discussed. PMID- 12755185 TI - Military nursing at the forefront: the Army Forward Surgical Team. AB - From World War II to current conflicts, history has validated the need for early surgical intervention to save lives and established the need for FSTs. Historically 10% to 15% of soldiers wounded in action require surgery to control hemorrhage and to provide stabilization sufficient for evacuation to a medical treatment facility where definitive care can be provided. Undoubtedly, many lives were saved in past conflicts because resuscitative surgery and care were available a short time after being wounded. The need for surgical stabilization for patients to survive a long evacuation was well-established during military operations such as Operation Desert Storm. Resuscitative surgery capability must be present on the battlefield, and this capability must be able to move with the units the FST is supporting. These two imperatives were the driving force of the creation of the US Army's FSTs: they are light, easily transportable, and able to keep pace with the supported units. FSTs are an integral and essential element in providing surgical care to save soldiers' lives on the modem battlefield. The US military has been involved in more operations and deployments since the end of the Cold War than in the preceding years. FSTs participated in many of these operations and have performed superbly. FSTs will continue to have a critical role in providing combat health service support to soldiers on the battlefield of the twenty-first century, and FST nurses will continue to be an indispensable asset. PMID- 12755186 TI - EMEDs and SPEARR teams: United States Air Force ready responders. AB - The right mix and unique talents of the medical personnel brought together by chance are undoubtedly the basis for the accomplishments of the EMEDs deployed to support Operation Houston and Operation Enduring Freedom, whereas creativity and flexibility definitely remain the keys to the success of these missions. It is with great confidence that we can proudly say, "Yes, we are ready to respond when called upon to provide the highest quality care for humanitarian missions and for our deployed military personnel." PMID- 12755187 TI - Navy nurse anesthetists at Fleet Hospital Five: the Desert Shield/Storm experience. AB - In 1990, the United States Navy deployed its first operational fleet hospital: "Fleet Hospital Five" in support of Operation Desert Shield/Storm. Within 2 weeks of notification, the 900 medical providers assigned to this medical facility, which was capable of providing major trauma surgery and critical care, were on their way to Al Jabayl, Saudi Arabia. This article discusses the unique characteristics of this facility and introduces the crucial role that nurse anesthetists play. The article also introduces several innovative ideas that were developed and tested to expand the capabilities of the hospital. PMID- 12755188 TI - Global presence: USAF aeromedical evacuation and critical care air transport. AB - Flight nursing, whether as an AE nurse or as a CCAT team member, is a demanding profession that extracts tremendous energy, competes with family and recreational time, and sets high expectations. On reflection, however, most crewmembers claim it is the most rewarding experience in their professional life. The opportunity to be a part of history, to provide care and transport to American servicemen and women in times of extreme need, and to accomplish the mission safely despite the circumstances and personal cost is an unparalleled experience and one that hold tremendous pride. PMID- 12755189 TI - Blue water nursing: the role of Navy nurses on board US Navy combatant ships. AB - The independent and autonomous nature of blue water nursing makes its practice exceptionally demanding, exciting, and rewarding for Navy nurses assigned to US Navy combatant ships. Whether serving on board an aircraft carrier or an amphibious assault ship, the lone critical care nurse is responsible for a diverse mix of nursing care ranging from community health to critical care. Delivering this care in an austere shipboard environment at sea, often in isolated locations without regular available support from shore-based medical facilities, defines the blue water aspect of this nursing practice. Due to this type of setting and the limited critical care nursing presence on board, the greatest professional challenge at sea is care of the critically ill and injured. Leadership and teamwork are essential. Navy Corpsmen play an integral role in the delivery of nursing care, and the nurse relies heavily on them. Because the nurse cannot do it all alone, taking the lead on providing training and clinical guidance to the Corpsmen is a key responsibility of the shipboard nurse. In this assignment, the critical care nurse also has a unique opportunity to make an impact the health and welfare of the ship's entire crew through wellness and prevention programs. Considering the personal and professional challenges and rewards of blue water nursing, most Navy nurses describe their tour of duty on board a combatant ship as the best assignment of their Navy career. PMID- 12755190 TI - Patient care in a biological safety level-4 (BSL-4) environment. AB - The greatest threats to America's public health include accidental importation of deadly diseases by international travelers and the release of biologic weapons by our adversaries. The greatest failure is unpreparedness because international travel and dispersion of biologic agents by our enemies are inevitable. An effective medical defense program is the recommended deterrent against these threats. The United States has a federal response plan in place that includes patient care and patient transport by using the highest level of biologic containment: BSL-4. The DoD has the capability to provide intensive care for victims infected with highly infectious yet unknown biologic agents in an environment that protects the caregiver while allowing scientists to study the characteristics of these new agents and assess the effectiveness of treatment. Army critical care nurses are vital in the biologic medical defense against unidentified infectious diseases, accidental occupational exposures, or intentional dispersion of weaponized biologic agents. Research that carefully advances healthcare using BSL-4 technology addresses the challenges of the human element of BSL-4 containment patient care, and BSL-4 patient transport enhances our nation's ability to address the emerging biologic threats we confront in the future. PMID- 12755191 TI - The Army chemical/biological SMART (SMART-CB) team: the nurse's role. AB - A chemical or biologic attack probably will be covert, rather than overt. Because presenting signs and symptoms may mimic minor nonspecific illnesses or naturally produced disease syndromes and may not appear for several days, it is likely that nurses in emergency rooms and primary care settings will be among the first to come into contact with victims of a chemical or biologic agent exposure. Early recognition, reporting, decontamination, self-protection, prophylaxis, and treatment are imperative. After the terrorist attack on September 11, 2001, the anthrax incidents, and the heightened publicity of chemical and biologic agent attacks brought on by media coverage, the need for highly trained and well prepared medical personnel has increased dramatically. Army nurses have led the way in training and expanding the capabilities of specialized medical response teams. Team members require ongoing training, state-of-the-art protective equipment and medical supplies, and constant practice to maintain the high state of readiness required to respond rapidly and effectively to chemical or biologic threats. Army SMART-CB nurses and their team members are well prepared to provide lifesaving care in highly contaminated areas. It is no longer a question of if but rather when and where the next attack will occur. PMID- 12755192 TI - Peace making/peace keeping missions: role of the U.S. Army nurse. AB - Somewhere in the world every month expert military critical care/trauma providers are working side-by-side with host nations to help them develop their trauma and disaster management systems. This article discusses the Air Force Medical System's (AFMS) mission to provide humanitarian and civic assistance, disaster response, and care of wartime injured. Within the context of this tripartite mission, the article introduces the AFMS's flagship international course, "Leadership Course in Regional Disaster Response and Trauma System Management," and highlights the that military critical care nurses play in this international effort. PMID- 12755193 TI - International trauma and disaster management. AB - Our world has literally changed around us. Based on this reality, the AFMS changed its' way of thinking when providing care to victims in disaster situations. This course is but one of several that are provided around the world that speak one common language, helping each other and caring for our patients the best way we know how. As Kofi Annan, Secretary General of the United Nations, eloquently stated: "We are all in the same boat. There are no safe islands. There is no dividing line between domestic and international crises. There is no them, only us." PMID- 12755194 TI - Maximizing quality of life in ill animals. PMID- 12755195 TI - Accuracy of heart rate obtained by auscultation in atrial fibrillation. AB - The accuracy of heart rate estimation by cardiac auscultation over a 15-second period, and the influence of clinical experience on accuracy were evaluated in a dog with chronic atrial fibrillation by test subjects of varying experience. Only 30% of all test subjects provided accurate heart rate estimates. Board-certified specialists, medicine residents, and experienced nurses were significantly more accurate in their estimates than surgery residents and students. Accurate estimates were provided by 12.5% of surgery residents and students, as opposed to 64% of the other test subjects. Auscultatory estimates of heart rate in atrial fibrillation may be significantly inaccurate, and under some circumstances they may not provide a sound basis for making clinical decisions. PMID- 12755196 TI - A comparison of owner management and complications in 67 cats with esophagostomy and percutaneous endoscopic gastrostomy feeding tubes. AB - Esophagostomy feeding tubes were placed in 46 cats. Percutaneous endoscopic gastrostomy (PEG) feeding tubes were placed in 21 cats. Owner management and complications and facility of use were evaluated retrospectively by review of medical records and owner survey. Both tube types were equally effective for maintenance of body weight, ease of owner management, and complication rates. All of 12 owners surveyed were comfortable with PEG tube management. Ninety-six percent of 24 owners surveyed were comfortable with esophagostomy tube management. The esophagostomy tube can be placed less invasively, without specialized equipment, making it an excellent alternative to the PEG tube. PMID- 12755197 TI - Evaluation of systemic and secretory IgA concentrations and immunohistochemical stains for IgA-containing B cells in mucosal tissues of an Irish setter with selective IgA deficiency. AB - Immunoglobulin A is the predominant secretory antibody at mucosal surfaces. In the dog, immunoglobulin A deficiency (IgAD) is characterized by low to absent serum IgA and normal to elevated serum immunoglobulin G (IgG) and immunoglobulin M (IgM) concentrations. However, studies comparing serum and secretory IgA in dogs have often documented a poor correlation, suggesting that serum concentrations should not be used to estimate mucosal secretion of this antibody. This report demonstrates the concurrent use of serum IgA, IgG, and IgM; secretory IgA (from bronchoalveolar lavage fluid); and immunohistochemical stains on bronchial and duodenal mucosa for IgA-containing B cells in a young Irish setter with recurrent respiratory and gastrointestinal signs. PMID- 12755198 TI - Fatal hemothorax following management of an esophageal foreign body. AB - A 10.8-year-old, spayed female toy poodle presented with an esophageal foreign body. The foreign body was removed endoscopically, and a gastrostomy tube was placed to provide nutritional support during esophageal healing. The gastrostomy tube was later removed by endoscopic retrieval of the bulb through the esophagus. Immediately afterward, the dog developed hemothorax and eventually died. It was determined that many small arterial branches were avulsed from the aorta. The involved sections of aorta histopathogically evidenced medial necrosis, which was believed to be related to a prior disruption of blood flow through the vasa vasorum. PMID- 12755199 TI - Fluoroscopic and endoscopic localization of an esophagobronchial fistula in a dog. AB - A 10-month-old, intact male Chihuahua presented for a 7-month history of regurgitation and coughing. Survey radiographs revealed a soft-tissue opacity within the distal esophagus. A contrast study confirmed the presence of an esophagobronchial fistula. Endoscopic removal of foreign material within the esophagus allowed for visualization of an esophageal diverticulum. Bronchoscopic examination revealed the presence of an abnormal opening in a right caudal tertiary bronchus. Passage of a cardiac guidewire was accomplished, confirming the presence of the fistula and aiding its subsequent surgical removal. PMID- 12755200 TI - Candida spp. urinary tract infections in 13 dogs and seven cats: predisposing factors, treatment, and outcome. AB - Records from 20 animals (13 dogs, seven cats) with Candida spp. urinary tract infections were reviewed. Six Candida spp. were isolated; Candida albicans was the most common isolate. Concurrent diseases or nonantifungal drugs administered within 1 month of isolation included antibiotics (n=16), corticosteroids (n=6), diabetes mellitus (n=4), nonurogenital neoplasia (n=3), and noncandidal urogenital disease (n=14). All animals had sources of local or systemic immune compromise that likely predisposed to infection. Of five animals with resolution of infection, three did not receive specific antifungal treatment. The authors conclude that correction of predisposing conditions is likely critical for management of Candida spp. urinary tract infection. PMID- 12755201 TI - Spinal arachnoid cysts in 17 dogs. AB - The medical records of 17 dogs diagnosed with spinal arachnoid cysts at North Carolina State University Veterinary Teaching Hospital were retrospectively examined to identify trends in signalment, history, neurological status, treatment, and short- and long-term prognosis. The typical case was that of a nonpainful, progressive ataxia frequently characterized by hypermetria and incontinence. Cysts typically occurred in the dorsal subarachnoid space at the first to third cervical vertebrae of young, large-breed dogs or the caudal thoracic vertebrae of older, small-breed dogs. Although 14 of 15 dogs treated surgically did well in the short term, long-term successful outcomes were achieved in only eight of the 12 dogs that were followed for >1 year. Significant predictors of good, long-term outcome were not identified; however, factors associated with a trend toward a good outcome included <3 years of age, <4 months' duration of clinical signs, and marsupialization as the surgical technique. PMID- 12755202 TI - Chlorambucil-induced myoclonus in a cat with lymphoma. AB - Chlorambucil is an alkylating agent commonly used in veterinary oncology for conditions including lymphoma. Chlorambucil neurotoxicity has been well recognized in human patients. Onsets of central nervous system signs, such as myoclonus, tremors, muscular twitching, agitation, and tonic-clonic seizures, have been reported in humans and laboratory animals treated with chlorambucil. This case of a cat with intestinal lymphoma represents the first veterinary patient reported to have chlorambucil-induced neurotoxicity. Neurotoxicity should be considered a potential side effect of chlorambucil therapy in veterinary patients. PMID- 12755203 TI - Astrocytomas in young dogs. AB - Primary brain tumors are not commonly reported in young dogs; however, they are the second most common cancer in children. Astrocytomas are the majority of these tumors. This report presents three cases of astrocytomas in young dogs, indicating a possible higher incidence than what is currently held. When suspected, it is recommended that biopsy or surgical excision be performed to further characterize and grade the tumor and, if appropriate, guide treatment. PMID- 12755204 TI - Combined dorsolateral and intraoral approach for the resection of tumors of the maxilla in the dog. AB - This paper describes in detail a combined dorsal and intraoral approach for maxillectomy for tumors involving tissues more caudal to the third premolar. The only intraoperative complication was that of blood loss, with six out of 20 dogs requiring a single unit of blood. Histopathologically clean margins were obtained in 14 of the 20 cases, with a recurrence rate of 50% in these dogs and a median time to recurrence of 24 months. This represents an improvement in outcome over previously reported studies, and the authors postulate this is due to the better exposure and access to the area afforded by the combined approach over the standard intraoral approach. PMID- 12755205 TI - Surgical removal of an intramedullary spinal cord foreign body granuloma in a dog. AB - A 2-year-old, spayed female, mixed-breed dog was presented for evaluation of a progressive asymmetric tetraparesis and cranial nerve deficits with a 3-week duration. Computed tomography showed a contrast-enhancing lesion along the left side of the junction of the medulla and the cervical spinal cord. An exploratory surgery determined the presence of an intramedullary lesion of the first cervical spinal cord segment. The mass was removed through a dorsal midline myelotomy. Microscopic examination identified a foreign body granuloma that contained a dense, anisotropic outer wall, supporting the conclusion that the foreign body was of plant origin. The dog recovered to a more improved ambulatory status than prior to surgery. PMID- 12755206 TI - Postoperative integrity of veterinary surgical gloves. AB - A multicenter, prospective study was performed to document the incidence of defective gloves postoperatively in veterinary surgery and to correlate defects with a variety of influencing factors. Gloves were collected after surgical procedures performed by the small animal clinical services at two veterinary teaching hospitals and one institution's student surgery laboratories. Gloves were evaluated for defects using electrical resistance testing. The overall incidence of glove defects was 23.3%. Significantly more defects occurred in gloves used for nonsoft-tissue procedures and in gloves worn on the nondominant hand. Eighty-four percent of all defects occurred in procedures lasting >60 minutes. No differences were detected in the brands of gloves used nor among surgeons of different experience levels. The individuals performing the surgery were not able to accurately predict the presence of a defect in their gloves. Surgeons should remain alert for possible glove defects and consider measures such as changing gloves every 60 minutes or double-gloving to minimize potential complications. PMID- 12755207 TI - Corporate greed. PMID- 12755208 TI - Radiologic case study. Benign cartilaginous neoplasm; chondroblastoma. PMID- 12755209 TI - K-wire fixation of displaced physeal injuries. PMID- 12755210 TI - Anatomic and radiographic analysis of arthroscopic tack placement into the anteroinferior glenoid. AB - Tack location within the anteroinferior aspect of the glenoid when performing simulated repairs of anteroinferior capsulolabral avulsions (Bankart lesions) was evaluated anatomically and radiographically. Arthroscopy was performed on six fresh-frozen cadaveric shoulders, and bioabsorbable tacks were placed through an accessory anteroinferior portal coming into the joint just above the subscapularis tendon using an outside-in technique. Tack location was studied after removal of all soft tissues. In addition to their position on the glenoid, the tacks were also evaluated for being partially or completely within bone. The tacks were recannulated with guide pins and anteroposterior, axillary, and en face glenoid radiographs of each specimen were obtained. This study provides quantitative data about the inferior placement limitations of the insertion angle and location of fixation devices within the anteroinferior glenoid through the anteroinferior accessory portal. PMID- 12755211 TI - The incidence of pathology detected by magnetic resonance imaging of the knee: differences based on the specialty of the requesting physician. AB - The usage patterns of magnetic resonance imaging (MRI) by orthopedic and nonorthopedic surgeons were studied. A sample consisting of the radiologist reports from all knee MRIs in a single year at our institution were reviewed. Studies to evaluate tumors or infections were excluded. Reports were classified as normal or demonstrating degenerative joint disease, meniscal tears, cruciate ligament pathology, collateral ligament pathology, focal chondral defects, chondromalacia patella, cysts, extensor mechanism dysfunction, intraosseous edema, or fractures. Six hundred eighteen reports were reviewed. The combined incidence of a normal study or one that found only degenerative joint disease was 45% for nonorthopedic surgeons and 27.6% for orthopedic surgeons (P<.00001). Given the higher incidence of normal findings in studies ordered by nonorthopedic surgeons, these physicians probably use MRI more for screening whereas orthopedic surgeons are more apt to use it for confirmation. Therefore, if clinical guidelines for using MRI are to be established, differences in use as a function of specialty must be acknowledged. PMID- 12755212 TI - Long-term results of derotational femoral varus osteotomy in Legg-Calve-Perthes disease: 26-year follow-up. AB - The long-term results of 30 patients (31 hips) who underwent derotational femoral varus osteotomy for Legg-Calve-Perthes disease are presented. Pain, leg-length discrepancy, Trendelenburg sign, and range of motion at the operated hip were examined clinically. Radiographic analysis included measurement of the Wiberg angle, epiphyseal index, acetabular index, and the Mose index. All were found to be satisfactory for patients in the good/fair category. Good/fair results were obtained in 27 (87%) of 31 hips according to Catterall's postoperative classification. Four patients were classified in the poor category due to severe restriction of movement and constant hip pain. Therefore, derotational femoral varus osteotomy is recommended for the treatment of patients with Legg-Calve Perthes disease. PMID- 12755213 TI - Atrophy of the multifidus muscle in patients with lumbar disk herniation: histochemical and electromyographic study. AB - A histochemical and electromyographic study of the lumbar multifidus muscle in 17 patients with L4-L5 lumbar disk herniation was performed. Electromyography (EMG) was recorded preoperatively with a needle electrode according to Haig's method. Biopsy specimens were obtained intraoperatively from the L5 band of the multifidus muscle on the affected and unaffected sides. Patients with lumbar disk herniation showed atrophy of type 1 and type 2 fibers with structural changes in the multifidus muscle at the involved level. Furthermore, patients with abnormal EMG results had severe muscle atrophy compared with patients with normal EMG results. PMID- 12755214 TI - Interscalene block for elective shoulder surgery. AB - This study assessed patient experiences with interscalene block anesthesia for elective shoulder surgery. Routine use of interscalene anesthesia was introduced at our institution in 1997. All patients who underwent interscalene anesthesia during the first 3 years of our experience with this regional anesthetic technique were asked to respond to an anesthesia-related questionnaire. Overall patient satisfaction with interscalene anesthesia was 87%, and success rate in achieving a complete motor and sensory block was 79%. The duration of pain relief postoperatively was 10.5 hours for patients with a successful block. Patients consistently reported that having an interscalene block was less painful than anticipated; 90% said they would have interscalene anesthesia again for shoulder surgery. This study demonstrates that interscalene anesthesia for elective shoulder surgery is safe and well accepted in this patient population. PMID- 12755215 TI - Erdheim-Chester disease. PMID- 12755216 TI - Operative stabilization of an unstable os acromiale in an adolescent football player. PMID- 12755217 TI - Superficial and deep posterior compartment syndrome following high tibial osteotomy for tibia vara in a child. PMID- 12755218 TI - Delayed neurologic injury due to bone graft migration into the spinal canal following scoliosis surgery. PMID- 12755219 TI - Intra-articular patellar dislocation. PMID- 12755220 TI - Childhood septic arthritis following treatment of spondylodiskitis from Serratia marcescens. PMID- 12755221 TI - Pulmonary fat embolism after total hip and total knee arthroplasty. PMID- 12755222 TI - The effect of bone graft extenders to enhance the performance of iliac crest bone grafts in instrumented lumbar spine fusion. AB - Allograft bone extenders are commonly used in spinal surgery to increase the available graft volume, thereby promoting and achieving a solid fusion mass. We report a single surgeon's use and early results of autologous bone graft and allograft demineralized bone matrix in 65 patients undergoing lumbar spinal fusion. Of the patients included in this study, 59 (91%) patients underwent surgical intervention for lumbar spinal stenosis, three (5%) patients had lumbar spondylolisthesis, two (3%) patients had stenosis, and one (1%) patient had bilateral spondylolysis. Forty-three (64%) women and 22 (36%) men were included in the study. The average patient age was 56 years (20-85 years, SD= +/- 16). Independent radiographic evaluation was performed. Each subsequent radiographic follow-up revealed increased improvement in average Lenke score and was statistically significant between the early (1 month) and recent (12 month) follow-ups. There were statistically significant changes in Lenke score between 1 month and 3 months follow-up (P<.01), between 3 months and 6 months follow-up (P<.001), and between 6 months and 12 months follow-up (P<.01). The gradual and constant increment of improvement in radiographic measurements in this preliminary series may indicate a positive effect of the use of bone graft extenders that may decrease the required amount of autologous bone graft. Bone graft extenders also may minimize the risks and complications associated with the harvesting procedure. PMID- 12755223 TI - Percutaneous treatment of long bone nonunions: the use of autologous bone marrow and allograft bone matrix. AB - Sixty-six patients with 69 "stiff" nonunions (no gross motion) of long bones were entered into a prospective study. The only therapeutic intervention was the percutaneous administration of a mixture of autologous bone marrow and allograft demineralized bone matrix on an outpatient basis. Sixty-one of the percutaneous treatments (88%) resulted in union at an average of 8.1 months (range: 2 months to 3 years). This method of treating nonunions is as successful as standard iliac crest autologous bone grafting and offers the distinct advantages of decreased morbidity, reduced costs, and shorter hospital stay. PMID- 12755224 TI - Interpositional arthroplasty (Jones technique) for the treatment of herniated lumbar disks: a modification of the soft posterior lumbar interbody fusion. AB - Sixty-six patients underwent posterior lumbar interpositional arthroplasty using a combination of calcium sulfate pellets, decompression bone, and autologous growth factors. Patients who underwent this modification of the soft posterior lumbar interbody fusion (PLIF) (Jones technique) were evaluated using clinical and radiographic ratings. The Jones technique produced significantly improved clinical pain scores in all patients, reducing back pain by 71% and leg pain by 90%. Furthermore, 93% of patients achieved at least 50% opacity of the disk space area. The Jones technique for the soft PLIF provided reduction of pain and stabilized the disk space height in addition to decreasing morbidity and cost. PMID- 12755225 TI - Bone marrow aspiration. PMID- 12755226 TI - Restoration of large bone defects using a hard-setting, injectable putty containing demineralized bone particles compared to cancellous autograft bone. AB - An injectable, hard-setting, calcium sulfate-based putty containing demineralized bone matrix particles (AlloMatrix II, Wright Medical Technology, Inc, Arlington, Tenn) was compared to autogenous cancellous bone graft to evaluate healing in a canine model. Area fraction of new bone, modulus of elasticity, and compressive strength of new bone were evaluated, as was radiographic and histologic healing. Bilateral defects were created in the proximal humeri, and each defect was implanted with either the putty or autogenous bone according to a randomized schedule. Dogs were euthanized at 6, 13, and 26 weeks. The area fraction, modulus of elasticity, and compressive strength of newly formed bone was not significantly different between the putty and autogenous bone at 6, 13, or 26 weeks. The putty had excellent handling characteristics, was biocompatible, and was as effective as autograft bone in achieving near complete bony restoration of a large, critical-sized defect. PMID- 12755227 TI - The effect of allomatrix injectable putty on the outcome of long bone applications. AB - Long bone defects due to nonunion or surgical excision of benign bone tumors result in areas that require rapid regeneration of local bone. This clinical and radiographic article details the results of a commercially prepared allograft demineralized bone matrix in patients with long bone voids or gaps. Of the 76 patients included for study, 41 (54%) patients were undergoing surgical intervention for removal of benign tumors or space-occupying lesions and 35 (46%) patients had long bone nonunions. AlloMatrix Injectable Putty (Wright Medical Technology, Inc, Arlington, Tenn) was used alone in 74 (97%) patients and in combination with bone marrow aspirate in two (3%) patients with tibial nonunion. Adjunctive strut allografts were used in three patients with humeral nonunion. The average time to follow-up for the combined population was 7 months (nonunion group 6 months; benign tumor group 7 months). At the most recent follow-up, radiographic evidence of the average percent of bone healing was 85.1% for the nonunion patient group and 93% for the benign tumor patient group. From this study, AlloMatrix Injectable Putty used as a bone void filler in long bone nonunions and benign tumors shows results equal to those historically reported for autograft and other materials. PMID- 12755228 TI - Osseous healing using injectable calcium sulfate-based putty for the delivery of demineralized bone matrix and cancellous bone chips. AB - Three formulations of injectable calcium sulfate-based putties containing demineralized bone matrix (DBM), 50% DBM/50% cancellous bone (CB) chips, and 30% DBM/70% CB were studied in canines. Four humeral defects per dog were implanted with one of each of the putty formulations while the fourth defect was left untreated. After 6 weeks, the dogs were euthanized. Radiographs and histology showed that the area fraction of new bone in the defects was greater for the three putty formulations than the untreated defects. The area of residual cancellous bone graft remaining in the defects was <10% in both CB putties. Residual calcium sulfate was not apparent in any of the histological sections. We conclude that fast-resorbing calcium sulfate-based putties are effective delivery means of bone graft materials for the successful restoration of bony defects. PMID- 12755229 TI - Resorption evaluation of a large bolus of calcium sulfate in a canine medullary defect. AB - New bone formation and resorption of a calcium sulfate bone graft substitute implanted in five canines were evaluated in this study. Healing was assessed radiographically at 2, 6, and 13 weeks. At 13 weeks, the dogs were sacrificed, and the humeri were retrieved. High-resolution contact radiographs of the isolated humeri were obtained and the bones were sectioned for histology. Radiographically, the calcium sulfate appeared to be completely resorbed and replaced by bone at 13 weeks. Histological findings suggest that a residual amount of calcium sulfate remained, which may continue to act as an osteoconductive scaffolding. No adverse inflammatory response was observed. PMID- 12755230 TI - Healing of large defects treated with calcium sulfate pellets containing demineralized bone matrix particles. AB - Calcium sulfate (OsteoSet, Wright Medical Technology, Inc, Arlington, Tenn) and calcium sulfate/demineralized bone matrix (DBM) pellets (OsteoSet DBM, Wright Medical Technology, Inc) have been evaluated preclinically in a bilateral medullary defect model of a canine humerus. In this model, both short (6 week) and long (26 week) time points have been evaluated. An analysis of bone response to the pellets was conducted using radiological, histological, mechanical, and quantification techniques. The calcium sulfate/DBM pellets exhibited more rapid trabecular bone remodeling as demonstrated by the absence of the ringlet bone structure typically seen with calcium sulfate pellets. We concluded that calcium sulfate and calcium sulfate/DBM pellets are both effective bone graft substitutes. PMID- 12755231 TI - The use of a periosteal replacement membrane for bone graft containment at allograft-host junctions after tumor resection and reconstruction with bulk allograft. AB - Bone grafting is frequently used to assist in reconstruction of bone defects following limb salvage procedures in tumor surgery. These defects can be large and require the use of bulk allograft and/or significant amounts of particulate graft material. This article reports a clinic's preliminary results with a bone graft containment material that serves as an artificial periosteum. This radiographic review evaluates the material's ability to prevent migration of bone graft and ascertain progressive callus formation. Early results have been encouraging. PMID- 12755233 TI - Contribution of vaccinations towards reducing morbidity and mortality among children in developing countries. PMID- 12755232 TI - The use of acellular dermal matrix as a scaffold for periosteum replacement. AB - Three preclinical models were used to evaluate GraftJacket Acellular Periosteum Replacement Scaffold (Wright Medical Technology, Inc, Arlington, Tenn). The studies assessed the ability of the acellular dermal matrix to repopulate with cells, revascularize, provide a protected environment for bone defect restoration, and minimize fibrous tissue infiltration. An athymic nude rat muscle implantation study demonstrated a steady increase in cellular repopulation through days 2-21. The formation of blood vessels occurred between days 7-14 in this study. Results from a porcine femoral drill hole study indicated that the scaffold material was intact and adherent to surrounding bone and allowed cellular repopulation and vascular infiltration at a 5-week time period. A preliminary porcine segmental bone defect model at a 6-week time period demonstrated the ability of the scaffold material to protect the bone defect site as revealed by new bone formation within the margins of the defect and adjacent to the scaffold. The segmental model also indicated minimal to no soft tissue invasion into the defect site. The combined studies provided preliminary evidence that the dermal membrane material may be used as a scaffold for periosteum regeneration by allowing for cellular repopulation, revascularization, and bone defect restoration. PMID- 12755234 TI - Comparison of vaccination status of children born in health units and those born at home. AB - BACKGROUND: The Expanded Programme of Immunisation schedule starts at birth, yet a significant number of child births in Uganda occur at home, where there are no vaccines. A child born at home may therefore have less chances of being vaccinated than a child born in a health unit. OBJECTIVES: To investigate vaccination status of the under-fives and to establish whether vaccination status of children born in health units is better than those born at home. DESIGN: Cross sectional descriptive. SETTING: Paediatrics out-patient clinic of Jinja Hospital, a regional referral health facility in Eastern Uganda. METHODS: Children under five years were enrolled consecutively as they arrived at the registration desk. The child health card and physical examination for BCG scar were used to establish the vaccination status of each child. A structured questionnaire was used for collecting relevant data. RESULTS: Of the 486 children sampled, 79 had been born at home, and 407 had been born in health units. Overall, 68% of the children were fully vaccinated for age. A child born in a health unit was significantly more likely to have a BCG scar (p = 0.0087), and to be up to date with their vaccination (p = 0.0173), compared to a child born at home. Vaccine drop-out rate was similarly high irrespective of whether the children were born at home or in health units. CONCLUSION: Being born at home was found to be a risk factor for incomplete or non-vaccination. Continuation of vaccination was similarly poor in children born at home and those born in health units. PMID- 12755235 TI - Primary health care, selective or comprehensive, which way to go? AB - OBJECTIVE: To critically review the advantages and disadvantages of selective versus comprehensive Primary Health Care (PHC) approaches as a strategy towards improving health in the developing world. DATA SOURCES: Review of literature on PHC. DATA SELECTION: Relevant papers from western and developing world literature. DATA EXTRACTION: Search of Pub-Med, WHO/UNICEF reports, and relevant publications on PHC. DATA SYNTHESIS: Examination of principles behind PHC and practical experiences in PHC in the developing world. CONCLUSIONS: Selective PHC programs have improved specific aspects of health, frequently at the expense of other health sectors, but fail to address an individual's health in holistic manner, or the health infrastructure of countries. Selective PHC programs tend to focus only on a small subset of the community. Comprehensive PHC is expensive to implement, however addresses health of individuals more holistically, addresses both preventive and curative health care, and promotes health infrastructure development and community involvement, thereby providing more sustainable improvement of health in the whole community. PMID- 12755236 TI - Daily versus weekly iron supplementation and prevention of iron deficiency anaemia in lactating women. AB - OBJECTIVE: To demonstrate the effectiveness and social feasibility of weekly versus daily iron supplementation in preventing and treating iron deficiency anaemia among anaemic mothers. DESIGN: A longitudinal in nature. SETTING: Seven urban slum communities in Teklehaimanot Wereda, Addis Ababa, Ethiopia. SUBJECTS: Two hundred seven eligible mothers were assigned to the daily supplementation, weekly supplementation or control groups following randomisation between March and May 2001. The daily supplemented groups (n=71) received 60 mg of elemental iron containing 300 mg ferrous sulphate and 400 microg folic acid from Monday to Friday. The weekly group (n=68) received one tablet once a week every Monday supervised while the control group (n=68) was advised to take no medications without the knowledge of the investigators until the completion of the study. To eliminate a major source of variation, subjects participating in the study were de-wormed at the beginning of the study. MAIN OUTCOME MEASURES: Haemoglobin and serum ferritin concentrations were compared before and after the intervention among the groups. RESULTS: The mean haemoglobin (Hgb), and serum ferritin concentration (SFC) at baseline were practically similar among the groups. Haemoglobin levels significantly increased at the end of the study in all the groups and the proportion of anaemia decreased from 6.9% to 1.6% in the daily, 6.7% to 1.7% in the weekly supplemented and 6.7% to 6.1% in the control groups. The difference noted between the daily and weekly supplemented groups was not significant. The improvement of SFC concentration was better in the daily than the weekly group but not statistically significant. Daily supplementation schedule caused more side effects and lower compliance level than the weekly supplementation schedule. CONCLUSION: Weekly supplementation is simple, comparable to daily supplementation and economically advantageous. Thus, it is recommended to adopt the strategy for controlling anaemia. Further because of higher compliance rate and lower side effects, it is deemed to be socially feasible. PMID- 12755237 TI - Vitamin A supplementation on child morbidity. AB - OBJECTIVE: To determine the impact of vitamin A supplementation on child morbidity and nutritional status. DESIGN: A community based follow-up (interventional) in nature. SETTING: Two randomly selected Weredas (districts) of Tigray, North Ethiopia were studied between 1996 and 1997. SUBJECTS: Four thousand seven hundred and seventy children aged between six and 72 months, selected using a multi-stage sampling procedure were enrolled and clinically assessed for xerophthalmia and nutritional status. A sub-sample of these children (n = 281) was further assessed for their serum retinol levels. MAIN OUTCOME MEASURES: The pre and post intervention data on xerophthalmia, morbidity, nutritional status and serum retinol levels were compared. RESULTS: Vitamin A capsule coverage of 87% in all the villages of the Weredas and a statistically significant (p < 0.05) reduction in the prevalence of Bitot's spot (from 1.5 to 0.5), fever (from 29.8 to 14.2), diarrhoea (from 30.2 to 18.2), oedema (from 9.2 to 3.2), measles (from 14.0 to 6.2), conjunctivitis (from 10.2 to 3.0), stunted (from 64.2 to 42.7), wasted (from 12.8 to 2.5) and underweight (from 46.2 to 24.2). The proportion of children with normal serum retinol concentration (> 0.7 micromole/L) has also improved significantly (from 36.8 to 56.2). CONCLUSION: In conclusion, the significant improvement in morbidity and nutritional status that followed the intervention programme although encouraging, it still indicates the importance of coupling periodic provision of Vitamin A capsules with nutrition education. PMID- 12755238 TI - Immediate outcome of babies with low Apgar score in Mulago Hospital, Uganda. AB - BACKGROUND: Birth asphyxia contributes significantly to perinatal morbidity and mortality especially in resource poor countries. Although the Apgar score has been in use for over 50 years, the prevalence of low Apgar score and attendant risk factors and outcome have not been established in many sub-Saharan countries including Uganda. OBJECTIVE: To determine the prevalence of low Apgar score and establish immediate outcome and possible risk factors for poor outcome in babies with low Apgar score. SETTING: Labour wards, operating theatres and special baby care unit, Mulago Teaching and referral Hospital, Uganda. SUBJECTS: Babies delivered in Mulago Hospital between September and October 1999. Those with low Apgar scores, together with an equal number of babies with normal scores matched for sex as controls, were followed up for 48 hours. MEASUREMENTS: Clinical features, anthropometry, gestational age, oxygen saturation, blood glucose and autopsy of babies who died. MAIN OUTCOME MEASURES: Clinical improvement, death, complications such as HIE, RDS, aspiration pneumonia, hypoglycaemia, hypothermia, hypotension and hypoxaemia. RESULTS: The prevalence of low Apgar score at one and five minutes was 8.4% and 2.8% respectively. Adverse outcome was seen in 57.3% of cases: death in 12.1% and clinical complications in 45.2%. HIE occurred in 21.8%, hypoxaemia in 12.9%, hypoglycaemia in 16.9% and aspiration pneumonia in 4.8%. Maternal factors significantly associated with low Apgar scores included primiparity, abnormal delivery, age and medical diseases during pregnancy, while birth injuries and cord accidents were the baby factors. Poor outcome was associated with birth injury, hypothermia, hypoglycaemia, hypotension, aspiration pneumonia, hypoxaemia and severe birth asphyxia. CONCLUSION: Even though the prevalence of low Apgar was only 8.4%, adverse outcomes associated with it were observed in more than half the patients. Therefore there is need to carefully evaluate and monitor babies with low Apgar scores immediately after birth. PMID- 12755239 TI - Pneumocystis carinii pneumonia in HIV/AIDS patients at an urban district hospital in Kenya. AB - BACKGROUND: Pneumocystis carinii pneumonia has generally been regarded to be an uncommon opportunistic infection in HIV infected individuals in sub-Saharan Africa. The reason for this has not been clear but postulates included a lack of suitable pathogenic types in the African environment, diagnostic difficulties and the more commonly held belief that African HIV infected individuals were dying early from common non-opportunistic pathogens before severe degrees of immunosuppression occured. Recently a trend has emerged at the Mbagathi district hospital whereby an increasing number of HIV infected patients are empirically treated for Pneumocystis carinii pneumonia (PCP) based on clinical and radiological features. OBJECTIVE: To determine the prevalence of PCP and clinical outcomes of HIV infected patients presenting at the Mbagathi District Hospital, Nairobi with the presumptive diagnosis of PCP. SETTING: Mbagathi District Hospital, a 169-bed public hospital in Nairobi, Kenya. METHODS: Patients presenting with a sub-acute onset of cough and dyspnoea were eligible for the study if they were found to have bilateral pulmonary shadows and had negative sputum smears for AFBS. Consenting patients who had no contraindication to fiberoptic bronchoscopy had a clinical evaluation which was followed with a fiberoptic bronchoscopy procedure where bronchoalveolar lavage fluid (BALF) was obtained. BALF was examined for cysts of P. carinii using toluidine blue stain and immunofluorescent antibody test (IFAT). BALF was also processed for fungi, bacteria and mycobacteria using routine procedures. Standard treatment with high dose cotrimoxazole was offered to all patients who were then followed up until discharge from hospital or death whichever came first. RESULTS: Between June 1999 and August 2000 a total of 63 patients were referred for bronchoscopy. Of these four declined to undergo the fiberoptic bronchoscopy procedure, four died before the procedure could be done, one was judged too sick to undergo the procedure and three had been on cotrimoxazole for longer than five days. Thus 51 patients underwent bronchoscopy. Pneumocystis carinii stain was positive in 19 (37.2%) while death occured in 16 (31.4%) of the 51 patients. There were more deaths in those without PCP but this difference was not statistically significant (odds ratio 0.68 (95% CI 0.35-1.32; P=0.2). CONCLUSION: PCP was found to be common in HIV infected patients presenting with clinical and radiological features of the disease. The mortality rate for patients with a presumptive diagnosis of PCP is high. This study suggests that cotrimoxazole preventive therapy may be a useful intervention in symptomatic HIV infected patients in Kenya for the prevention of PCP and may avert deaths from this disease. PMID- 12755240 TI - Risk factors and prevalence of diabetic foot ulcers at Kenyatta National Hospital, Nairobi. AB - BACKGROUND: Diabetic foot ulcers contribute significantly to the morbidity and mortality of patients with diabetes mellitus. The diabetic patients with foot ulcers require long hospitalisation and carry risk of limb amputation. The risk factors for developing diabetic foot ulcers are manageable. In Kenya there is paucity of data on such risk factors. OBJECTIVE: To determine the prevalence of diabetic foot ulcers and the risk factors in a clinic-based setting. DESIGN: Cross-sectional study. SETTING: Kenyatta National Hospital, Kenya. SUBJECTS: Patients with both type 1 and 2 diabetes mellitus who had active foot ulcers in both outpatient and inpatient units. MAIN OUTCOME MEASURES: Diabetic foot ulcers glycated haemoglobin, neuropathy, peripheral vascular disease and fasting lipid profile. RESULTS: One thousand seven hundred and eighty eight patients with diabetes mellitus were screened and 82 (4.6%) were found to have foot ulcers. The males and females with diabetic foot ulcers were compared in age, duration of foot ulcers, blood pressure, glycaemic control, neurological disability score and their proportion. Diabetic foot ulcers occurred mostly in patients who had had diabetes for a long duration. The types of (occurence) ulcers were neuropathic (47.5%), neuroischaemic (30.5%) and ischaemic (18%). The neuropathic ulcers had significantly poorer glycaemic control compared to other types and the longest duration (23.3 weeks). Ischaemic ulcers had significantly higher total cholesterol and diastolic blood pressure compared to other ulcer types. Wagner stage 2 ulcers were the commonest (49.4%) but stage 4 ulcers had their highest neuropathic score (7.8/10) and longest duration (23.6 weeks). Aerobic infective pathogens were isolated from 73.2% of the ulcers. CONCLUSION: The prevalence of diabetic foot ulcers was 4.6% in this tertiary clinic. The risk factors of diabetic foot ulcers in the study were poor glycaemic control, diastolic hypertension, dyslipidaemia, infection and poor self-care. These findings are similar to studies done in other environments and they are modifiable to achieve prevention, delay in formation or improved healing of foot ulcers in patients with diabetes. Therefore, specific attention should be paid to the management of these risk factors in patients with or without diabetes foot ulcers in this clinic. PMID- 12755241 TI - Experience with laparoscopic surgery at the Aga Khan Hospital, Nairobi. AB - OBJECTIVE: To outline the experience of laparoscopic surgery at the Aga Khan Hospital, Nairobi, and to determine the acceptability and outcome of the various procedures undertaken laparoscopically. DESIGN: A retrospective case analysis. SUBJECTS: Four hundred and seventy eight cases of laparoscopic surgery were undertaken in Nairobi from May 2000 to May 2002 in the presence of the principal author, of these 408 cases (85.35%) were performed at the Aga Khan Hospital in Nairobi. A review of these cases and their outcomes are analysed and presented. EXCLUSIONS: Loss of client records, or follow up, all diagnostic procedures and all procedures performed at the three other private hospitals, namely; Nairobi Hospital, Nairobi Womens' Hospital and the M.P. Shah Hospital, have been excluded in this review. RESULTS: Minimal access surgery will in time find its place amongst the institutions in Kenya. In the last two years 408 cases were undertaken at the Aga Khan Hospital, Nairobi, involving a pool of 48 consultants obstetricians and gynaecologists and general surgeons. The case load increased from 7.0 cases per month in 2000 to 22 cases per month in 2001. Sixty four point four six percent of the patients spent one night in hospital, while 1.96% had conversion to laparotomy intraoperatively. Thirty nine point seven percent of the patients had no previous surgery. The surgical procedures performed included laparoscopic adhesiolysis (34.55%) and tuboplasty (17.89%) for primary or secondary infertility, 33 cases (8.08% ) for the management of ectopic pregnancies, laparoscopic myomectomy (15.44%), ovarian cystectomy (16.91%), ovarian drilling (4.65%), laparoscopic assisted vaginal hysterectomy (15.19%) and total laparoscopic hysterectomy (2.20%). All cases were reviewed by the consultant pool one week after discharge. The major complications encountered included bladder injury (0.49%) and gut injury (0.73%). CONCLUSIONS: Minimal access surgery in gynaecology and general surgery is gaining remarkable ground worldwide and has tremendous potential in Kenya. It is evident that in trained hands, the common gynaecological operations and certain general Surgical procedures can be undertaken safely, laparoscopically. Minimal access surgery is acceptable to the patients and significantly favourable outcomes have been established in all the cases undertaken, thus far. Patient compliance has been excellent in this series. PMID- 12755242 TI - Induction of labour using low and high dose regimens of prostaglandin E2 vaginal tablets. AB - OBJECTIVE: To compare the efficacy of low dose prostaglandin E2 vaginal tablets with that of high dose in induction of labour. DESIGN: A retrospective study. SETTING: Abha Maternity Hospital, a teaching hospital in the southern region of Saudia Arabia. PATIENTS: The outcome of induction of labour in 73 women induced with 1.5 mg prostaglandin E2 vaginal tablets was compared with those in 168 women induced with 3 mg vaginal tablets. RESULTS: There were no statistical significant differences in the success rate, induction-delivery interval, caesarean section rate and Apgar scores between the two groups. There were two incidences of uterine hyperstimulation and two of uterine rupture in the high dose group. These were not statistically significant. Logistic regression analysis showed that the success rate of induction of labour was dependent only on maternal age (P=0.0025) and Bishop score (P=-0.0403) and not on parity, gestational age, birthweight or dose of prostaglandin. CONCLUSION: The low dose regimen of 1.5 mg is as efficacious as the high dose regimen of 3 mg prostaglandin E2 vaginal tablets in inducing labour. This implies that using the low dose regimen reduces the cost of induction of labour effectively. Larger prospective randomized studies are needed to confirm this finding. PMID- 12755243 TI - Polycystic kidney disease in a patient with achondroplasia: case report. AB - Autosomal dominant polycystic kidney disease is a multisystem disease involving many organs. An association with other diseases such as tuberous sclerosis, von Hippel-Lindau disease and Marfan syndrome have been previously described. We describe a 35 year old female with achondroplasia who developed polycystic kidney disease involving both kidneys and progressing to end-stage renal disease. To the best of our knowledge this is the first such case described in the literature. We also delve, briefly, into the possibility of the genes and chromosomes involved in Marfan syndrome, polycystic kidney disease, tuberous sclerosis and achondroplasia playing a role in the co-occurrence of these entities. PMID- 12755244 TI - Size, course, distribution and anomalies of the middle cerebral artery in adult Nigerians. PMID- 12755245 TI - Effectiveness of interventions to increase Papanicolaou smear use. AB - BACKGROUND: Many women fail to adhere to Papanicolaou smear screening guidelines. Although many interventions have been developed to increase screening, the effectiveness of different types of interventions is unclear. METHODS: We performed a systematic review of interventions to increase Papanicolaou smear use published between 1980 and April 2001 and included concurrently or randomized controlled studies with defined outcomes. Interventions were classified as targeted to patients, providers, patients and providers, or health care systems and as behavioral, cognitive, sociologic, or a combination based on the expected action of the intervention. Effect sizes and 95% confidence intervals were calculated for each intervention. RESULTS: Forty-six studies with 63 separate interventions were included. Most interventions increased Papanicolaou smear use, although in many cases the increase was not statistically significant. Behavioral interventions targeted to patients (eg, mailed or telephone reminders) increased Papanicolaou smear use by up to 18.8%; cognitive and sociologic interventions were only marginally effective, although a single culturally specific, sociologic intervention using a lay health worker increased use by 18.0% (95% confidence interval [CI]: 7.6, 28.4). Provider-targeted interventions were heterogeneous. Interventions that targeted both patients and providers did not appear to be any more effective than interventions targeted to either patients or providers alone. One of the most effective interventions, which introduced a system change by integrating a nurse-practitioner and offered same-day screening, increased screening by 32.7% (95% CI: 20.5, 44.9). CONCLUSIONS: Overall, most interventions increased Papanicolaou smear use, although there was tremendous variability in their effectiveness. Selection of intervention strategies will depend on provider and patient population characteristics and feasibility of implementation. PMID- 12755246 TI - LEEP in the family practice setting. AB - BACKGROUND: We wanted to review our 7-year experience using the loop electrical excision procedure (LEEP) for the treatment of cervical dysplasia in a family practice residency setting in the rural South. METHODS: We conducted a retrospective study with data gathered from chart review of a mostly Medicaid and uninsured patient population of rural Southern women referred from outlying health departments or private practices within an 80-mile radius. The women received follow-up Papanicolaou smears, and outcome measurements were either recurrence of dysplasia or at least 1 year with two negative Papanicolaou smears. Any surgical tissue obtained after LEEP was used to ascertain residual or recurrent dysplasia. RESULTS: Rates of disease recurrence and incomplete excision of cervical intraepithelial neoplasia grade 2/3 (CIN 2/3) compared favorably with results published by expert US gynecologists but were worse than those reported by European authors, who excise all CIN (CIN 1, CIN 2, and CIN 3). CONCLUSION: CIN 2 and CIN 3 can be diagnosed and treated appropriately with LEEP in the setting of a family practice residency. PMID- 12755247 TI - Physician prescribing patterns of oral corticosteroids for musculoskeletal injuries. AB - BACKGROUND: There is considerable controversy as to when and if oral corticosteroids are indicated for musculoskeletal injury. Although the perception is that sports medicine physicians prescribe oral corticosteroids frequently, there is no documentation of this behavior in the literature. The purpose of this study was to obtain a description of the corticosteroid-prescribing patterns of primary care sports medicine physicians and look for common indications. METHODS: A two-page questionnaire was included in the registration packets of 195 physicians at a national sports medicine conference. RESULTS: Ninety-nine questionnaires were returned. Fifty-eight (58.6%) of the physicians reported prescribing oral corticosteroids for musculoskeletal injuries. Physicians who prescribed corticosteroids for injuries averaged 6.6 prescriptions per month. Prednisone was the corticosteroid prescribed by 82% of physicians. The average prescription length was 7 days. One half of the physicians (51.7%) tapered the dose. The most common starting dose (mode) was 60 mg. CONCLUSIONS: Despite little evidence to support their use, primary care sports medicine physicians commonly prescribe corticosteroids. PMID- 12755248 TI - Precipitants of constipation during early childhood. AB - BACKGROUND: Childhood constipation is a common problem, accounting for 3% of visits to pediatric clinics and 30% of visits to pediatric gastroenterologists. Estimates of the prevalence of childhood constipation vary from 0.3% to 28% with younger children being affected most often. We were unable to find any studies that specifically examine the causes of constipation in young children. Our objective of the study was to determine precipitants to constipation during early childhood. METHODS: Findings from 125 families visiting their primary care physician for the first time with a child aged between 2 and 7 years with the complaint of constipation were compared with findings from 95 children between 2 and 7 years without any history of constipation. Parents answered questions concerning family history, toilet training, and bowel habits. Parents of constipated children were asked to describe events that occurred during the 3 months before the onset of constipation and whether these events contributed to the child's constipation. RESULTS: The age and sex of children who did and did not suffer from constipation were comparable (P > .3). When compared with control children, constipated children were no more likely to have a parent (30% vs 40%, P = .14) or sibling (17% vs 14%, P = .54) with a history of constipation. Constipated children did not begin toilet training earlier than did control children (28 +/- 7 vs 27 +/- 6 months, P = .30). When compared with parents of control children, parents of constipated children reported more difficulties with toilet training (P < .001). Parents of constipated children indicated their children had more difficult and more painful defecation experiences than did parents of control children (P < .001), and constipated children were more likely to express worry about future painful defecation than were control children (P < .001). Parents of constipated children described a number of events that occurred before the onset of constipation; however, they did not consider many of the events important contributors to the constipation. Painful defecation was the event most often reported as causing the constipation. CONCLUSION: Painful defecation is the primary precipitant of constipation during early childhood. Parents should be counseled to be attentive to such experiences and taught to intervene quickly to lessen the risk that their child will develop persistent constipation or fecal soiling. PMID- 12755249 TI - Attaching a new understanding to the patient-physician relationship in family practice. AB - BACKGROUND: As a result of continuity of care with patients and their families, family physicians are uniquely poised to form enduring clinical relationships with their patients. The degree of collaboration in and satisfaction with the patient-provider alliance has been shown to have important implications for treatment outcomes across a range of medical problems. Providing optimal care can require family physicians to appreciate the sequelae of having clinically relevant aspects of past relationships emerge in the health care relationship, both in their patients and in themselves. A conceptual model is essential to assist in recognizing these key aspects. METHODS: A literature search was conducted using MEDLINE. Key words entered were "illness" and "attachment theory." Thirty-five English-only articles appeared from which further relevant references were gathered. RESULTS: Attachment theory serves as a useful model for highlighting important features of physician-patient relationships, which can affect treatment outcome in the family practice setting. It posits that everyone has an innate need to form strong attachment bonds to their earliest caregivers. To ensure survival, the child adapts its bonding to the caregiver's attachment style. With time, the maturing person develops a style of relating in subsequent caregiving relationships based on these early, and to some extent later, close relationships. Insecure attachment styles that can develop--dismissing, preoccupied, and fearful--have been shown to affect the clinical relationship and medical treatment outcomes often in important and predictable ways. CONCLUSION: Family physicians can more easily adopt an understanding, compassionate, and flexible treatment stance by recognizing patients' unique attachment relationship patterns, thereby improving medical treatment outcome. PMID- 12755250 TI - Computer-based testing in family practice certification and recertification. AB - BACKGROUND: The member boards of the American Board of Medical Specialties have agreed to expand the scope of certification to include assessment of medical knowledge, practice-based learning and improvement, patient care, interpersonal and communication skills, systems-based practice, and professionalism. Multiple choice examinations provide limited ability to assess these dimensions. METHODS: The American Board of Family Practice (ABFP) has developed a computer simulation system to facilitate more comprehensive candidate evaluation. The system consists of a knowledge base, a simulation program to create patient scenarios, an interface for presenting simulations to users, and an administrative database to track candidate performance and interactions with the system. The system uses population distributions for disease states to produce cases and evolves patients in response to candidate interventions, such as pharmacological and nonpharmacological therapies. We use Bayesian belief networks to model patient characteristics and comorbid condition interactions. RESULTS: Simulations have been created for 7 disease states; ultimately simulations will be available for 25 to 30 disease states. Initial testing will take place in regional examination centers but will ultimately use the Internet for convenient access for certification and recertification candidates. CONCLUSION: The ABFP will begin field-testing the system in early 2003 and will include simulations in the certification and recertification examination process in 2004. PMID- 12755251 TI - Breast cancer screening controversies. AB - BACKGROUND: The Cochrane Collaborative, a respected independent review body, recently published a meta-analysis of the effectiveness of screening mammography in decreasing breast cancer mortality. Based on the results of two controlled trials they judged to be of medium validity, they concluded that screening mammography was unjustified. In contrast, the US Preventive Services Task Force recently updated their screening recommendations, and based on a meta-analysis of the same randomized controlled trials, they recommended screening mammography for all women starting at age 40 years. Additionally the Canadian Task Force on Preventive Health Care no longer recommends breast self-examination (BSE). This article reviews the controversies regarding breast cancer screening. METHODS: We performed a systematic review of the literature using keywords and cross referencing articles. We also used automated data from the Breast Cancer Screening Program at Group Health Cooperative to determine the sensitivity of the clinical breast examination (CBE) at our institution. For the latter we included all cancers diagnosed within 1 year of a screening examination and then determined which of those had been found by CBE. RESULTS: Although most screening studies have shown that mammography decreases breast cancer death, there are controversies about the validity of some of the randomized controlled screening mammography trials. These controversies have led to different conclusions about the efficacy of screening mammography. Evidence is limited about the optimal interval for screening mammography. No studies have directly tested the efficacy of the CBE in decreasing breast cancer mortality. At Group Health Cooperative, 8% of all diagnosed breast cancers were found by the CBE alone (negative mammogram). Whether this 8% incremental increase in case finding leads to decreased breast cancer deaths is unknown. There is good evidence that training women to perform BSE does not increase breast cancer diagnoses or decrease breast cancer deaths. CONCLUSION: There are limitations to randomized controlled trials and meta analyses. The balance of the evidence still favors screening mammography in women aged 40 years and older at least every 2 years. The independent incremental benefit of the CBE, when added to mammography, in decreasing breast cancer mortality is unknown. Population-based education and training to do BSE are unlikely to lead to decreased breast cancer deaths. Many women find their own breast cancers, so women need to pay attention to symptoms or changes in their breasts. PMID- 12755252 TI - A physician's guide to working as a locum tenens. AB - BACKGROUND: The number of physicians working as a locum tenens is increasing. Although most physicians who provide locum tenens coverage are often older and semiretired, an increasing number of physicians are taking this route for limited periods early in their careers. METHODS: The medical literature was searched through MEDLINE using the key words "career choice," "contract services," and "locum tenens." Information about locum tenens was gained by the author through research and a personal experience working as a locum tenens. RESULTS AND CONCLUSION: Working as a locum tenens can be an opportunity to meet interesting patients, learn about local culture, see how practices are organized, learn adaptability, and broaden clinical skills. Opportunities are available through for-profit agencies, academic health centers, state agencies, federal sites such as the Indian Health Service, individual hospitals, and physicians or international staffing companies. Because the physician working as a locum tenens is an independent contractor, exercising some caution when choosing where to work by carefully checking workload and available resources is advised. PMID- 12755253 TI - When deep venous thrombosis fails to respond to therapy. AB - BACKGROUND: Deep venous thrombosis in primary care is usually treated with rest, analgesics, intravenous or low-molecular-weight heparin, and coumadin. In some cases, however, a less familiar course of diagnosis and management is required. METHODS: We describe the case of a 53-year-old truck driver who had an acute deep venous thrombosis of his right lower extremity, which failed to respond to routine therapy with heparin and warfarin. A literature search was undertaken to research the differential diagnosis and management of deep venous thrombosis and to review specifically the role of venal caval filters and inherited thrombotic disorders and occult cancer in this context. RESULTS AND CONCLUSION: The ultimate diagnosis in our patient appeared to be signet ring cell cancer of the colon that had metastasized to the right thigh. This case is an example of the inherent limitations of even an aggressive diagnostic and therapeutic approach to the entity of refractory deep venous thrombosis. PMID- 12755254 TI - Type 2 diabetes mellitus induced by an atypical antipsychotic medication. PMID- 12755255 TI - An elderly woman with severe anxiety associated with anticipated use of an interpreter. PMID- 12755256 TI - Why incremental reforms will not solve the health care crisis. PMID- 12755257 TI - Medical student reflects on hospice experience. PMID- 12755258 TI - Staying connected to hospice patients. PMID- 12755259 TI - Study on the accuracy of automated hematology analyzers in Shanghai. AB - The objectives of this study were to research on the accuracy of automated hematology analyzers of various types from different manufacturers and to observe the deviation among these instruments. Fresh anticoagulated blood from healthy donors on 115 hematology analyzers in 114 different hospitals were determined. RESULTS: The maximum coefficients of variation (CVs. %) among instruments of three main manufacturers (Sysmex, Beckman Coulter and Abbott) of red blood cell count (RBC), hemoglobin (Hgb), hematocrit (Hct), white blood cell count (WBC) and platelet count (Plt) were 3.2%, 3.8%, 3.6%, 9.3% and 10.8%, respectively. The maximum deviations among these parameters of different instruments were 0.74%, 1.65%, 5.45%, 7.06% and 18.55%, respectively. By improving laboratory quality management, the results of hematology analyzer determination may be more reliable than manual methods. The difference among various manufacturers was very small about RBC, Hgb, Hct, WBC and Plt the results from all kinds of instruments will tend to be comparable. PMID- 12755260 TI - Hematology tests of blood anticoagulated with magnesium sulphate. AB - Venous blood treated with magnesium sulphate (MgSO4) at blood collection was used for hematology tests. Complete blood counts and automated leukocyte differentials were obtained using a hematology analyzer, and the results obtained for blood treated with MgSO4 were similar to those for blood treated with dipotassium ethylenediaminetetraacetic acid (EDTA K2). Coagulation tests such as activated partial thromboplastin time, platelet factor 3 availability and prothrombin time were dose-dependently affected by the use of MgSO4. Thrombin time was prolonged by addition of MgSO4, and fibrin concentrations determined by coagulometry tended to decrease with addition of MgSO4, although fibrin concentrations determined by the weighing method were unaffected by MgSO4. MgSO, was thus found to potently inhibit blood coagulation and can be used as an anticoagulant for hematology tests. PMID- 12755261 TI - The present status of the clinical laboratory medicine in Cambodia. AB - The educational system and the introduction of legislation of clinical medicine are both still in developing stage in Cambodia where only 10 years have passed since the establishment of a new government. In order to maintain good health of all Cambodian citizens and to improve the quality of care in health services, it should be necessary to implement an appropriate educational system for both laboratory technologists and technicians. To conduct refreshment training course for laboratory workers with provision of the instruments, material and reagents is another way to make improvement of it in public hospitals. It should be also required to overcome economic problems how to absorb medical expense and to understand the importance for doctors to diagnose with scientific data of clinical examinations. Maturation of the total medical system in this country should be necessary and suggestions from neighboring countries with views toward the world standard would be expected. PMID- 12755262 TI - Thailand intensive external quality assessment schemes as dynamic tools for improving laboratory quality and standard. AB - The term 'external quality assessment (EQA)' has been recommended by WHO consultative meeting in 1980. It has been using to run the retrospective and com parative statistic tests' reliability among the EQA participants of their organizers. EQA schemes in all fields of laboratory pathology should provide objective information as tools to assist laboratory analysts ensuring the test result quality and on the contrary deciding error-eradicated actions, or more or less improving national test performance quality and standard. Scoring assessment has been designed in the EQA schemes to assist monitoring, and encouraging quality and standard of laboratory performance in the long run of consecutive trials. The Faculty of Medical Technology of Mahidol University now as a collaborating WHO:SEARO EQA organizers, has provided wider EQA schemes, servicing participants all around Thailand, including 400 labs in clinical chemistry of EQAC scheme, 150 labs in clinical hormone of EQAH scheme, 200 labs in clinical microscopy of EQAM scheme, and 190 labs in clinical immunology and serology of EQAI schemes. Regular EQA newsletters, and yearly summer-time seminar and training prove to be the essential education tools promoting both laboratory quality and standard intensively. PMID- 12755263 TI - Proposed evaluation of laboratory performance in an external hematology quality control scheme. AB - The commercial control material for hematology Cell Dyn 16 Tri Level gave a good precision an accuracy using the four blood cell counters currently mostly used in Indonesia, the Coulter, Sysmex, Serono and Cell Dyn. It could be used as one of the hematology control material in an external quality control scheme. Result of the CV from participating laboratories are higher compared to this trial are caused by different level of laboratory technical ability although the geographical area and climate may also play a role. The new scoring calculation to evaluate the participant's performance gave a more wider range of DI scores, to give better insight to the performance of each laboratory. PMID- 12755264 TI - Problems on national and locale external quality assurance schema (NEQAS and LEQAS); based on Japanese experience. AB - The quality and stability of surveillance materials is a critical issue to perform good external quality assessment schema. In the case of nationwide surveillance, more stable materials are required than those in the case of locale external quality control schema. For locale surveillance, fresh blood, plasma or serum materials are prepared for hematology and chemistry tests. On the other hand, dried materials are used for the chemistry test and semi-fixed blood is used for hematology tests in the national surveillance. However to evaluate automated white cell differential in hematology, there has been no good materials. We therefore prepared a mixture of EDTA and ACD solution for our locale surveillance. It gave us excellent results on the differential when we used it within 3 days after blood collection. PMID- 12755265 TI - Experience of the first survey of the pilot project of Asian Quality Assurance Survey program. AB - The Asian Network for Clinical Laboratory Standardization (ANCLS) decided to start her First Inter-laboratory Quality Assurance survey at the business meeting during the Second Asian Colloquium on October 21-22, 2000 in Kobe, Japan. The first survey materials of Asian Quality Assurance Survey (AQuAS) were distributed in July 2001 to 21 hospitals in the field of chemistry and 22 hospitals in the field of hematology among total 24 participating laboratories from seven Asian countries: Indonesia, Japan, Korea, Malaysia, Philippines, Singapore and Thailand. The survey methods in chemistry and in hematology were somewhat different. In chemistry the survey material was considered as unknown but handled similarly to the sample from the patient. Nineteen routine tests were performed only once. The hematology samples were considered as unknown but handled similarly to control or calibration material. Five parameters were tested five times repetitively and their average for each parameter was calculated. All the results were supposed to be sent back to the office within two weeks. Although it took more than two weeks, the return rate was 100%. The analysis was performed in several ways such as all the results together, by instruments and by methods. Mean, standard deviation (SD), standard deviation index (SDI), coefficient of variation (CV) and variance index score (VIS) were to be calculated in chemistry, and in hematology the same parameter were to be calculated except CV and VIS. In the first survey, the CV in chemistry was not calculated and the analysis by instrument or by methodology was also not attempted since there were not enough participating hospitals to do such analysis. In hematology the analysis was done by instrument only. The survey process was carried out successfully though there were some difficulties in communication tools, transportation methods and handling of specimens due to different weather conditions, and returning the report in the correct unit and to the correct place. The submitted data were acceptable for analysis. There were some differences in the units of measurement in different countries or laboratories. It was necessary to convert some of the units. Some laboratories apparently do not perform certain tests such as calcium, potasium and gamma-glutamyltransferase (gamma-GT). The gamma-GT is the most frequently not performed test. With the experience of this first survey, all the members involved in the survey have been trained well to do future surveys. PMID- 12755266 TI - The Indonesian society of clinical pathologist external quality assessment program (ISCP-EQAP) in clinical chemistry. PMID- 12755267 TI - Blood bank quality assessment program in Singapore. PMID- 12755269 TI - Comparison of three Dade-Behring Dimension AR machines in a clinical chemistry laboratory. AB - Three Dade-Behring Dimension AR Analyzers are presently in use in the Philippine General Hospital Department of Laboratories. One machine was acquired 2 years ago and two new ones were added recently. In order to determine whether the three machines would produce equivalent results 16 samples were selected for patients with hepatic and/or renal disorder. All 16 samples were tested for 16 different analytes. The results were subjected to tests for coefficient correlation, simple linear regression using slope and intercept and t-test for comparison. We concluded that for two assays (globulin and phosphorus), the values were sufficiently similar. For seven other assays (for total protein, albumin, sodium, potassium, chloride, calcium, and magnesium), the results were statistically significantly different, and for the remaining eight assays (total hiliruhin, direct bilirubin, AST, ALT, ALP, glucose, BUN, and creatinine), the values obtained were not ideal for drawing up conclusions about comparability. For the latter group of assays we plan to pool patient specimens of appropriate numbers and levels and repeat testing at some specified later date. PMID- 12755268 TI - Second national external quality assessment scheme in hematology: the Philippine experience. AB - The Philippine Council for Quality Assurance In Clinical Laboratories has conducted two National External Quality Assessment Schemes (NEQAS) in Hematology. The first survey was conducted in December 1999 and the second in August 2000, with 95 and 187 laboratories, using mostly automated analyzers, participating respectively. Control materials were distributed during a two-week period by human network, and analyzed over a six to eight week period. For the first survey, only 36 laboratories (38.0%) submitted results. Data was divided into 4 peer groups based on the manufacturer. Since most of the samples were hemolysed upon analysis, only WBC and HGB parameters were evaluated. No outliers were detected in each peer group after analysis by the 'Peer Group Mean and SDI' method. Using the clinical laboratory improvement act of 1988 proficiency testing criteria (CLIA'88), only 5 results (13.9%) were unsatisfactory for WBC, and all results were satisfactory for HGB. For the second survey, 87 laboratories (47%) responded. Data was divided into 5 peer groups. There were few incidents of sample deterioration. Although majority of the coefficient of variations were acceptable, about 23 (12.6%) participants showed abnormality in at least one parameter after analysis by the 'Peer Group Mean and SDI'. Using CLIA'88, 5 WBC (6.5%), 6 RBC (7.6%), 8 HGB (9.7%), 15 HCT (19.0%), and 7 PLT (8.0%) results were unsatisfactory. In summary, the first NEQAS study served as a pilot study. Valuable lessons were learned for the improvement of the second NEQAS. The second NEQAS study was marked by a much larger sample size and better results. PMID- 12755270 TI - Evaluation of hematological values obtained with reference automated hematology analyzers of six manufacturers. AB - We evaluated assays of the same fresh blood samples with six different types of reference automated hematology analyzers developed by the following manufacturers: Beckman Coulter, Sysmex, Bayer, Abbott, Nihon Kohden and Horiba. Fresh whole blood samples treated with dipotassium ethylenediaminetetraacetic acid (EDTA K2) were collected from three healthy adult volunteers. The complete blood counts (CBC) including red blood cell count (RBC), hemoglobin (Hgb), hematocrit (Hct), mean corpuscular volume (MCV), white blood cell count (WBC), platelet count (Plt), reticulocyte percentage (Ret) and leukocyte differential counts including % neutrophils (Neu), % lymphocytes (Lym) and % monocytes (Mon) were surveyed with a reference automated hematology analyzer from each manufacturer. The process from sampling to analysis was performed according to procedures in hospital clinical laboratories. RBC, Hgb, Hct and MCV exhibited allowable differences within 5% of mean value among all instruments. Large differences greater than 10% of mean value in WBC, Neu and Lym between Horiba and other manufacturers, and in Plt between Nihon Kohden and other manufacturers, were observed. Ret and Mon exhibited large differences over 10% of mean value among almost all of the instruments tested. This survey suggests that all parameters exhibiting differences greater than 10% of mean value among instruments should be improved for clinical use to ensure good external quality control in blood cell counting and leukocyte differential counting using automated instruments. PMID- 12755271 TI - Three-year experience in using total laboratory automation system. AB - We implemented a total laboratory automation (TLA) system, currently used in Chungnam National University Hospital (CNUH), designed around the A&T Corporation to perform the general and specific laboratory testing throughout the system in a timely and cost-effective manner. The system consists of two major lines: chemistry and hematology. The analyzers attached to the hematology line include one SE 9000 hematology analyzer (Sysmex Corporation, Kobe, Japan), one SP-100 slide maker (Sysmex), and one R9000 reticulocyte counter (Sysmex). The chemistry line consists of preanalytic system and linked analyzers. The analyzers attached to chemistry line are one Hitachi 747 chemistry analyzer (Hitachi, Ltd., Tokyo, Japan), one A&T 502X analyzer (A&T Corporation, Tokyo, Japan) and one Architect i2000 immunochemistry analyzer (Abbott Diagnostics Division, Santa Clara, USA). The preanaytic system including start stocker, centrifuge, de-cap unit and aliquoting unit were connected with analyzers. The turn around time (TAT) of all tests was shortened in comparison to manual systems. Consequently it makes feasible one-day care clinic and makes reduced inherent errors in sample identification, dispensing, and reporting. In addition, real-time re-run can make test quality more reliable. As many as 12 departments can be reduced to 7 departments. The subsidiary advantages of smaller size were incorporated into reduced working steps facilitating workflow. Overall, better qualified and quicker results can be provided to the clinicians, and proper cost-effectiveness and better quality services can be anticipated as a result of much easier system management and higher reliability. In conclusion, the TLA has been successful within our laboratory. PMID- 12755272 TI - The development of systematic quality control method using laboratory information system and unity program. AB - Quality control (QC) process is performed to detect and correct errors in the laboratory, of which systematic errors are repeated and affect all the laboratory process thereafter. This makes it necessary for all the laboratories to detect and correct errors effectively and efficiently. We developed an on-line quality assurance system for detection and correction of systematic error, and linked it to the Unity Plus/Pro (Bio-Rad Laboratories, Irvine, USA), a commercially available quality management system. The laboratory information system based on the client-server paradigm was developed using NCR3600 (NCR, West Columbia, USA) as the server and database for server was Oracle 7.2 (Oracle, Belmont, USA) and development tool was Powerbuilder (Powersoft Burlignton, UK). Each QC material is registered and gets its own identification number and tested the same way as patient sample. The resulting QC data is entered into the Unity Plus/Pro program by in-house data entering program or by manual input. With the implementation of in-house laboratory information system (LIS) and linking it to Unity Plus/Pro, we could apply Westgard's multi-rule for higher error detection rate, resulting in more systematic and precise quality assurance for laboratory product, as well as complementary to conventional external quality assessment. PMID- 12755273 TI - Accreditation of clinical laboratories in the Philippines. AB - In order to assure the reliability of the results of laboratory services, clinical laboratories are regulated in the Philippines. This started with the passage of the Clinical Laboratory Law in 1965, which required the Licensing of clinical laboratories by the Bureau of Research and Laboratories, Department of Health (BRL, DOH) before they can operate. Standards were set for the various types of laboratory services. In 1988, the minimum standards of laboratory services were formulated for three categories, ie, primary, secondary and tertiary categories. Subsequently, to permit clinical laboratories to offer 'special service', accreditation of clinical laboratories was instituted. In 1968, the Philippine Society of Pathologists (PSP) decided to accredit clinical laboratories for Residency Training Program in Anatomic and Clinical Pathology. The following year after the passage of the Medical Technology Law, the BRL, DOH began accrediting clinical laboratories that trained Medical Technology Interns. A few years later, the BRL, DOH started to accredit clinical laboratories who did Water Analysis. In 1989, after realizing the serious implications of HIV Testing, the DOH mandated the BRL, DOH to set standards for clinical laboratories performing HIV testing. In 1997, upon request of the Philippine Health Insurance Corporation (PhilHealth), the PSP formulated and submitted standards for the accreditation of clinical laboratories, both hospital and free-standing, for reimbursement of fees for laboratory services rendered to patients enrolled in the PhilHealth social insurance program. In 2000, the Philippine Council for Accreditation of Healthcare Organizations (PCAHO) approved the Standards for the accreditation of Hospitals for the provision of quality medical services. Included were the standards for the Department of Pathology. PMID- 12755274 TI - Strategic plan and action plan for standardization and harmonization of hematology laboratories in Thailand. PMID- 12755275 TI - Clinical pathologist in Korea--training program and its roles in laboratories. AB - A rapid development of practice of laboratory medicine in Korea owes its success to the clinical pathologists (CP), who have played a role of a pathfinder for laboratories. The Korean CP postgraduate education (residency) program is unique in that it is exclusively for laboratory medicine. The training program for clinical pathologists includes diagnostic hematology, diagnostic immunology, clinical microbiology, clinical chemistry, blood bank, diagnostic genetics, informatics and laboratory management. The program has produced a strong group of about 600 laboratory physicians, officially clinical pathologists since 1963. Most of Korean clinical pathologists work as laboratory directors, directors of university hospital laboratories or teaching faculty members in medical schools. The roles of clinical pathologists are laboratory management, interpretation of laboratory test results, clinical consulting services to clinicians and patients, ordering secondary tests after reviews of requested test results and utilization management. The clinical pathologists have developed clinical laboratories to be a main contributor for improved medical practice. During the last 40 years under the turbulent healthcare system, clinical pathologists have significantly contributed to safeguard the laboratory interests. The education program and the role of clinical pathologists are described. PMID- 12755276 TI - External quality assurance schemes in flow cytometry. AB - Quality assurance schemes were devised to monitor the performance of laboratory tests commonly utilized in routine clinical practice. In the area of Flow Cytometry, external quality assurance schemes are very much in their infancy. There are 3 major applications of Flow Cytometry in the clinical laboratory: lymphocyte subset enumeration, CD34 enumeration and leukemia immuno-phenotyping. Pilot schemes for CD34 enumeration and leukemia immuno-phenotyping have revealed a considerable lack of consensus with a wide divergence in practices. The coefficient of variation in reported results from these assessments have been very large, ranging from 20-200%. Results for lymphocyte subset enumeration surveys are sometimes not much better despite having been around for a longer period of time. Quality assurance programs in Flow Cytometry are difficult to conduct and 3 or 4 concurrent factors are often cited as reasons for the large variability. The need for viable, functionally intact cells as suitable assay material is probably the biggest contributor to the large variability. This difficulty in providing stable cells for inter-laboratory comparison is compounded by the lack of consensus in testing methodology. Participation in a distant external quality assurance scheme is consequently not very helpful to improving the practices in the laboratory. Currently, there are more problems than solutions. An awareness of these issues will hopefully contribute to the development of future consensus and offer up solutions to some of these problems. PMID- 12755277 TI - External quality assessment scheme for blood coagulation. AB - The aim of setting an External Quality Assessment Scheme (EQAS) for Blood Coagulation is to achieve a harmonization among the participating laboratories on blood coagulation testing. In Indonesia EQAS for blood coagulation is organized by the Department of Clinical Pathology, Faculty of Medicine, University of Indonesia which cooperates with the Indonesian Society of Clinical Pathologists, Jakarta Chapter. Currently coagulation tests in Indonesia are only performed in a limited number of laboratories especially in the hospital. Therefore only 65 laboratories participated in the first trial of EQAS. The control material for EQAS was provided by Organon Teknika and parameters involved in the first trial were PT, INR, APTT, and fibrinogen. Currently there are 7 brands of reagents for coagulation tests available in the market, i.e.: Behring, Biomerieux, Biopool, Human, Nycomed, Organon, and Ortho. In the evaluation, the results of each participant were compared to the median of participants who used the same reagent. If the number of participants using a given reagent was less than 10, then the result of each participant was compared to the median of overall participants. The result of a given parameter was classified as within consensus if it fell into the range of median +/- 15%. In the overall evaluation, the percentage of participants which was classified as within consensus for PT, INR, APTT, and fibrinogen were 68%, 64%, 63%, and 67%, respectively, but the CV for PT, INR, APTT, and fibrinogen were 19.84%, 17.89%, 20.21%, and 23.96% respectively. In the evaluation of participating laboratories using Behring's reagent, the percentages of participants classified as within consensus for PT, INR, APTT, and fibrinogen were 72%, 82%, 54%, and 74%, respectively. For Organon's product users, the percentages of those parameters were 84.6%-90%, 48% 75%, 86%, and 67%, respectively. It is concluded that around two-thirds of overall participating laboratories achieved harmonization in the results of coagulation test, but the variation of all parameters is still wide. PMID- 12755278 TI - Quality control and quality assessment of coagulation tests in Japan. AB - We have used prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (Fbg), and fibronogen/fibrin degradation products (FDP) as screening coagulation tests, and antithrombin III (AT III), plasminogen (Plg), alpha2 plasmin inhibitor (alpha2-PI), protein C (PC), thrombin-AT III complex (TAT), plasmin-alpha2-PI complex (PIC), and D-dimer as special coagulation tests. We report the present condition of internal and external quality control for these coagulation tests in Japan. We made a summary report of internal quality control of some coagulation tests in Division of Hematology Laboratory, Department of Laboratory Medicine, Hokkaido University Hospital. The repeatability and reproducibility of screening tests were good, but those of special tests were partly adequate. We have participated in some external quality control surveillance including Japan Medical Association (JMA), Japanese Association of Laboratory Medical Technologists (JALMT), College of American Pathologists (CAP), and some commercial quality control surveillance. We also reported the results of some external quality control surveillance. PMID- 12755279 TI - External quality assurance in diagnostic immunology: a twenty-year experience in Korea. AB - External Quality Assurance (EQA) Program in diagnostic immunology was founded in 1982 in Korea, starting from proficiency testing for HBsAg and serological tests for syphilis. Proficiency testing for antisteptolysin O (ASO) and Widal tests was started in 1983, rheumatoid factor (RF), C-reactive protein (CRP) and anti-HBs tests in 1986, anti-HIV in 1992, and anti-HCV in 1993, now assessing total nine immunological tests. EQA surverys are performed twice annually and about 400 laboratories have been participating now. Over 75% of participating laboratories have used immunoassays including enzyme immunoassay for HBsAg, anti-HBs, anti-HCV and anti-HIV, and less than 20% of laboratories used hemagglutination tests. Overall error rate for HBsAg was less than 5%, those for anti-HBs and anti-HCV about 3% and anti-HIV less than 0.5%. Only negative samples are now used for anti HIV proficiency testing and two levels (negative and weakly positive or positive) of samples should be included in proficiency testing to assure the results. Thirty-five to forty-five percentages of participating laboratories have used nephelometry or turbidimetric immunoassay for CRP, RF and ASO. When comparing two kinds of nephelometry reagents most popularly used in Korea, quantitative results for CRP and RF by one kind were statistically different from the other. The reason for these discrepancies was not clear yet, however, standardization should be required. In the future, EQA in diagnostic immunology in Korea not only includes laboratory performance evaluation, but also evaluation of method performance and reagent evaluation. PMID- 12755280 TI - Histocompatibility testing. AB - The human leukocyte antigen (HLA) region on chromosome 6p21.3 is the most polymorphic in the human genome. It encodes hundreds of genes, of which the class I and class II HLA alleles play a central role in the generation of an immune response, but at the same time represent a barrier to marrow and organ transplantation. There is overwhelming evidence that optimal HLA matching will allow for safer marrow transplants, however with more than 1,200 allelic variants, the need to accurately identify the precise alleles is a technically demanding challenge. Methods currently in use for HLA typing include microcytotoxity assay, sequence-specific oligonucleotide (SSO) probe, sequence specific primer (SSP) amplification, and sequence-based typing (SBT). Ambiguous results may still arise, even with the best molecular typing methods but this problem can be resolved by using a combination of methods. It is clear that a HLA typing laboratory will have to provide allele level resolution of HLA data using the most appropriate means available and this will have the greatest impact in terms of offering the best matched donor for the benefit of patients. PMID- 12755281 TI - Evaluation of the stability of Cell Dyn 16 Tri Level as a control material for laboratory external quality assessment scheme on hematology. AB - The implementation of a laboratory test should always implement a laboratory quality control program, i.e internal quality control and external quality assessment. In an external quality assessment scheme, a control material that is stable over delivery until tested by the participating laboratory. In this study, we evaluated the stability of Cell Dyn 16 Tri Level (TL) control material at room temperature (26-32 degrees C), stored in a transport vessel containing ice pack, and the precision and accuracy of the instrument Cell Dyn 1400. The control used was Cell Dyn 16 TL with low value (L), normal value (N) and high value (H). This study was done in the Clinical Pathology Department of FKUI-RSCM during February 2001 until May 2001. Control material was stored room in a transport vessel containing ice pack for 15 days, then analysed macroscopically, microscopically and evaluated for its stability. Test for precision and accuracy was done within run and for precision between day on Cell Dyn 1400. The result of this study showed a macroscopic change beginning on day 14 (L) day 12 (N) and day 15 (H). Microscopic change was observed on day 13 (L and N) and day 15 (H), Erythrocyte and hemoglobin level was stable until day 15. Changes in leukocyte was seen on day 14 (L), day 12 (N) and day 15 (H). Platelet showed instability on day 9 (L), day 10 (N and H). Mean erythrocyte volume was out of range on day 15 (L), but the N and H control was still stable. The precision and accuracy of Cell Dyn 1400 was in WHO recommended range. We concluded that the precision and accuracy of Cell Dyn 1400 is good. Cell Dyn 16 TL control material was stable until day 9, and its can be recommended to be used as a control material for external quality assessment scheme. PMID- 12755282 TI - Serum levels of bone-specific alkaline phosphatase and procollagen type I carboxyterminal peptide in vitamin D deficiency. AB - Vitamin D deficiency in adults causes osteomalacia where there is a defect in bone mineralization resulting in an excess of unmineralised osteoid in the bone matrix. The aim of this study was to evaluate the markers of bone formation: total (TALP), bone-specific alkaline phosphatase (BSALP) and procollagen type I carboxyterminal peptide (PICP) in vitamin D deficiency. We studied 100 vitamin D deficient subjects and 82 gender-matched controls. Vitamin D deficiency was defined as serum 25-hydroxyvitamin D level of less than 7 ng/ml, and greater than 10 ng/ml for normal controls. Serum TALP assay was performed by a standard automated method, BSALP and PICP were measured by enzyme immunoassays (Metra Biosystems) and vitamin D by radioimmunoassay. There was significant difference in the TALP between female vitamin D deficient and control subjects (mean +/- sem = 99.8 +/- 8.2 vs 70.5 +/- 2.8 iu/l, p<0.001). Elevated serum TALP (>130 iu/l) was found in 20% (20/100) of the vitamin D deficient patients. There were no significant differences in BSALP or PICP between vitamin D deficient patients and gender-matched control subjects. There was no correlation between vitamin D and PICP in patients but in control subjects, a significant negative correlation (r= 0.431, p<0.0001) was found. In conclusion, although elevated TALP was observed in a minority of vitamin D deficient patients, it is a better marker than PICP. The lack of PICP response in vitamin D deficient subjects suggests the possibility of vitamin D deficiency leading to a block in osteoblast differentiation. PMID- 12755283 TI - Application of indicators for quality improvement in the coagulation laboratory. AB - Indicators are tools that measure work performance and serve as a guide to improve the quality of laboratories. Seven Indicators for quality improvement have been established in our coagulation laboratory. They are :- 1). percentage of pre-analytical problems, 2). personnel competency scores, 3). results of external quality assessment, 4). % coefficient of variation (CV) of control materials, 5). unit cost, 6). percentage of reports within determined time, and 7). percentage of customers who were satisfied. The percentage of preanalytical error gradually decreased from 1.8% in April 2001 to 0.8% in June 2001 as a result of co-operation between the coagulation laboratory and the wards. Since there is no system to check personnel competency at a national level in Thailand, we set up a program for testing personnel competency in our department by asking every technician to take a written and practical laboratory examination. The scores achieved by our personnel ranged from 40 to 90%. For those who achieved scores of lower than 70%, we limited their responsibilities and organized a training program for them. In order to check our laboratory's accuracy, we are enrolled in the WHO International External Quality Assessment Scheme (IEQAS) in Blood Coagulation and have been since 1987. The survey results indicated that most of our laboratory tests were within consensus including our homemade ELISA tests for protein C, protein S and vWF antigen. The percent CVs of control materials used for the internal daily control for every test were analyzed. They ranged from 2.3 for normal APTT to 11.4 for the low level of free protein S in plasma. The unit cost for each test was analyzed to determine the cost effectiveness of the laboratory. We set the goal for the turn around time for emergency coagulation tests to be within an hour and the percentage of reports within this time was 91.6% in August 2001. The last indicator was the percentage of satisfied customers, which gave an indication of the quality of all Out Patient Department (OPD) services performed by our department. We sent 400 questionnaires to doctors, nurses and patients in OPD asking their opinion of both the technical services and the behavior of our technicians. The percentage satisfaction of our customers concerning services offered to OPD was lower than 50%. We plan to improve the last 2 indicators by expanding the space of the OPD/emergency laboratory and reorganizing the service system. All indicators mentioned above have helped to improve the quality of our laboratory greatly. PMID- 12755284 TI - Reference method for platelet enumeration. AB - The main principle of enumerating platelets in automated hematology analyzer is electric resistance system. However, there have been an increasing number of instruments that can enumerate platelets by optical system. The use of a flow cytometry (FCM) employing monoclonal antibody has been under study for the enumeration of platelets in recent years. At its meeting held in April 2000, the International Society for Laboratory Hematology (ISLH) decided on a protocol to study the possibility of using, as reference method, a monoclonal antibody employed FCM of enumerating platelets. In our present study, we obtained platelet count by the monoclonal antibody-employed method proposed by ISLH, and compared the result with platelet count obtained with an automated hematology analyzer XE 2100 (Standard counter at Scientific Division, Sysmex, Kobe, Japan) employing electric resistance system, and another platelet count obtained by optical system. PMID- 12755285 TI - Screening for the carriers of thalassemias and abnormal hemoglobins at the community level. AB - Thalassemia and abnormal hemoglobins are common genetic disorders in Southeast Asia. Thalassemia is not only an important public health problem but also a socio economic problem of many countries in the region. The approach to deal with the thalassemic problem is to prevent and control births of the new cases. This requires an accurate identification of couple at high risk to have a thalassemic child. The diagnosis of thalassemia carriers need several tests that are not practical for screening the population at large. In this study we used two simple laboratory tests to screen for potential thalassemia carriers and hemoglobin E individuals. Three-hundred pregnant women and 40 spouses were recruited in this study. The methods were the red cell osmotic fragility (OF) screening test with 0.36% NaCl and the dichlorophenolindophenol (DCIP) precipitation test to detect Hb E and unstable hemoglobins. Standard methods for red cell indices, hemoglobin analysis and detection of alpha-thalassemia by immunological method were also performed to confirm genotypes of thalassemia. The results showed that 98 women (32.7%) were carriers of thalassemias and hemoglobin E. The number of false positive by OF test was 3.2% and by DCIP test was 0.6%. Sensitivity and specificity of OF test were 89.5% and 93.3%, respectively whereas those of DCIP test were 100%. Of the 40 couples investigated, one was found to be at risk of having beta-thalassemia/Hb E fetus. Screening techniques including one tube osmotic fragility and DCIP precipitation tests are sensitive and specific for the detection of thalassemia and unstable hemoglobins such as Hb E. The techniques are also simple, economic, rapid, and give minimal false negative result. PMID- 12755286 TI - Problems related to rapid methods for erythrocyte sedimentation rate test and their solution. AB - The erythrocyte sedimentation rate (ESR) test is one of the most common and traditional laboratory tests in the world. It reflects both plasma concentration of acute-phase proteins of large molecular size and anemia (International Council for Standardization in Hematology, 1993). The ESR test method is easy to perform and inexpensive. Therefore, it is used today as a routine test worldwide. However, the ESR has some demerits, in requiring a large volume of sodium citrate or ethylenediamine tetraacetic acid (EDTA) blood and at least 2-hour testing time. When ESR is tested manually, incorrect results are obtained if the ESR test tube is not stood strait-up and vertically (Dobashi et al, 1994). Reading error for the meniscus line and surrounding temperature at the testing site cause inaccuracy (Manley, 1957). The 2-hour testing time is not practical for modernized automated laboratories. Its test procedures present the risk of infection from contact with pathogen-bearing blood (Imafuku and Yoshida, 2001). In this context, several kinds of simple, rapid and safe methods have been developed. Of these new systems, we selected 4 and evaluated their performance. This paper reports critical reviews of such devices. PMID- 12755287 TI - A mini-review of CD13 antigen in AML: easy induction or enhancement of expression in in vitro culture and necessary consideration for assessment. AB - CD13 is a pan-(MPO-positive) AML antigen, while it is expressed in some B-lineage neoplasms (ALL and PLL) and in T-linaege neoplasms at pro-thymic stage. Some reports have described the lack of this antigen in MPO-positive AML, and tried to regard such MPO-postive AML cases as an clinical entity. However, considering the easy induction or enhancement of the expression in in vitro culture, it is pertinent to interpret that the expression of CD13 in these "CD13-negative and MPO-positive AML" cases is marginally positive, which is readily induced or enhanced in expression in in vitro culture. It can, however, be pointed out that the CD13 expression in ex vivo-positive cases are significantly stronger than that in ex-vivo-negative in vitro-positive cases. Such consideration is necessary particularly in interpreting the results obtained after overnight transportation in commercial laboratories. PMID- 12755288 TI - Prevention and control of influenza. Recommendations of the Advisory Committee on Immunization Practices (ACIP). AB - This report updates the 2002 recommendations by the Advisory Committee on Immunization Practices (ACIP) on the use of influenza vaccine and antiviral agents (CDC. Prevention and Control of Influenza: Recommendations of the Advisory Committee on Immunization Practices [ACIP]. MMWR 2002;51 [No. RR-3]:1-31). The 2003 recommendations include new or updated information regarding 1) the timing of influenza vaccination by age and risk group; 2) influenza vaccine for children aged 6-23 months; 3) the 2003-2004 trivalent inactivated vaccine virus strains: A/Moscow/10/99 (H3N2)-like, A/New Caledonia/20/99 (H1N1)-like, and B/Hong Kong/330/2001-like antigens (for the A/Moscow/10/99 [H3N2]-like antigen, manufacturers will use the antigenically equivalent A/Panama/2007/99 [H3N2] virus, and for the B/Hong Kong/330/2001-like antigen, manufacturers will use either B/Hong Kong/330/2001 or the antigenically equivalent B/Hong Kong/1434/2002); 4) availability of certain influenza vaccine doses with reduced thimerosal content, including single 0.25 mL-dose syringes; and 5) manufacturers of influenza vaccine for the U.S. market. Although the optimal time to vaccinate against influenza is October and November, vaccination in December and later continues to be strongly recommended A link to this report and other information regarding influenza can be accessed at http://www.cdc.gov/ncidod/diseases/flu/fluvirus.htm. PMID- 12755289 TI - Regional risk of exporting cattle seropositive for bluetongue virus from the United States. AB - OBJECTIVE: To create a stochastic model to quantify the risk that shipments of cattle from regions within the United States would contain animals seropositive for bluetongue virus and to determine shipment-level accuracy of serologic testing by use of a competitive ELISA (c-ELISA). SAMPLE POPULATION: 19,216 shipments containing 528,918 cattle and calves. PROCEDURE: Data were obtained on number of animals and state of origin of cattle in export shipments originating within the United States between January 1994 and March 2002. Probability distributions for size of export shipments were determined for all states within the United States, and distributions for agar gel immunodiffusion and c-ELISA accuracy (sensitivity and specificity) were determined from expert opinion and review of the literature. The model simulated selection of a shipment and then determined the probability that a threshold number or percentage of cattle within that shipment would have a positive c-ELISA result. Shipment-level sensitivity, specificity, positive-predictive value, and negative-predictive value were calculated. RESULTS: Substantial differences were evident in the regional probability of a shipment being declared positive, with shipments from northeastern states having the lowest probability and shipments from southwestern states having the highest probability. The c-ELISA had variable predictive values at the shipment level, depending on the threshold used and the prevalence of antibody-positive cattle within the region. CONCLUSIONS AND CLINICAL RELEVANCE: Results from this study will aid importers in making scientifically based decisions regarding risk of importing antibody-positive cattle. PMID- 12755290 TI - Prevention of persistent infection in calves by vaccination of dams with noncytopathic type-1 modified-live bovine viral diarrhea virus prior to breeding. AB - OBJECTIVE: To determine the ability of a modified-live virus (MLV) bovine viral diarrhea virus (BVDV) type 1 (BVDV1) vaccine administered to heifers prior to breeding to stimulate protective immunity that would block transmission of virulent heterologous BVDV during gestation, thus preventing persistent infection of a fetus. ANIMAL: 40 crossbred Angus heifers that were 15 to 18 months old and seronegative for BVDV and 36 calves born to those heifers. PROCEDURE: Heifers were randomly assigned to control (n = 13) or vaccinated (27) groups. The control group was administered a multivalent vaccine where-in the BVDV component had been omitted. The vaccinated heifers were administered a single dose of vaccine (IM or SC) containing MLV BVDV1 (WRL strain). All vaccinated and control heifers were maintained in pastures and exposed to BVDV-negative bulls 21 days later. Thirty five heifers were confirmed pregnant and were challenge exposed at 55 to 100 days of gestation by IV administration of virulent BVDV1 (7443 strain). RESULTS: All control heifers were viremic following challenge exposure, and calves born to control heifers were persistently infected with BVDV. Viremia was not detected in the vaccinated heifers, and 92% of calves born to vaccinated heifers were not persistently infected with BVDV. CONCLUSIONS AND CLINICAL RELEVANCE: These results document that vaccination with BVDV1 strain WRL protects fetuses from infection with heterologous virulent BVDV1. PMID- 12755291 TI - Neutrophil function and plasma opsonic capacity in colostrum-fed and colostrum deprived neonatal kittens. AB - OBJECTIVE: To determine whether passive transfer of IgG in neonatal kittens affects plasma opsonic capacity and neutrophil phagocytic and oxidative burst responses to bacteria in vitro. ANIMALS: 22 kittens from 6 specific pathogen-free queens. PROCEDURE: Kittens were randomized at birth into the following treatment groups: colostrum-fed, colostrum-deprived, or colostrum-deprived supplemented with feline or equine IgG. Blood samples were collected at intervals from birth to 56 days of age. Plasma IgG concentrations were determined by radial immunodiffusion assay. Neutrophil function was assessed by a flow cytometry assay providing simultaneous measurement of bacteria-induced phagocytosis and oxidative burst. The opsonic capacity of kitten plasma was determined in an opsonophagocytosis assay with bacteria incubated in untreated or heat-inactivated plasma. RESULTS: Among treatment groups, there were no significant differences in neutrophil phagocytic and oxidative burst responses to bacteria or opsonic capacity of plasma. In all samples of plasma, inactivation of complement and other heat-labile opsonins significantly reduced the opsonic capacity. Plasma IgG concentrations in kittens did not correlate with neutrophil function or plasma opsonic capacity before or after inactivation of complement. CONCLUSIONS AND CLINICAL RELEVANCE: The plasma opsonic capacity and neutrophil phagocytic and oxidative burst responses in vitro of kittens receiving passive transfer of IgG via colostrum intake or IgG supplementation and those deprived of colostrum were similar. The alternate complement pathway or other heat-labile opsonins may be more important than IgG in bacterial opsonization and phagocytosis. PMID- 12755292 TI - Development of a multiple-marker polymerase chain reaction assay for detection of metastatic melanoma in lymph node aspirates of dogs. AB - OBJECTIVE: To develop a reverse transcriptase-polymerase chain reaction (RT-PCR) assay to detect canine melanoma-associated antigens (MAAs) and to use this technique to screen aspirates of lymph nodes (LNs) for evidence of metastatic spread of oral malignant melanoma. ANIMALS: 7 dogs with oral malignant melanoma and 4 dogs with multicentric lymphosarcoma. PROCEDURES: We prepared cDNA from melanoma tumor biopsies and fine-needle aspirates obtained from submandibular LNs of dogs with oral malignant melanoma or multicentric lymphosarcoma. The RT-PCR assay was performed by use of tyrosinase, Melan-A, gp100, tyrosinase-related protein 2 (TRP-2), or melanoma antigen-encoding gene B (MAGE-B)-specific primers. RESULTS: We detected MAGE-B mRNA in canine testicular tissue but not in melanoma biopsy specimens. Tyrosinase, Melan-A, gp100, and TRP-2 mRNAs were detected in tumor biopsy specimens and in 2 of 5 LN aspirates from dogs with melanoma, suggesting metastatic spread in those 2 dogs. We did not detect MAAs in LN aspirates obtained from dogs with multicentric lymphosarcoma. Sequencing of canine Melan-A and gp100 PCR products confirmed the specificity of the assay for these genes. CONCLUSIONS AND CLINICAL RELEVANCE: Clinical staging of dogs with oral malignant melanoma is useful to assist in designing appropriate treatments. However, results of histologic examination of LN biopsy specimens can be inconclusive and, in humans, can underestimate the number of patients with metastatic disease. Molecular staging of melanomas in dogs can be achieved by screening LN aspirates for MAA mRNA, and this can be performed in combination with cytologic examination to aid in detection of metastatic disease. PMID- 12755293 TI - Coughing, mucus accumulation, airway obstruction, and airway inflammation in control horses and horses affected with recurrent airway obstruction. AB - OBJECTIVE: To investigate relationships between cough frequency and mucus accumulation, airway obstruction, and airway inflammation and to determine effects of dexamethasone on coughing and mucus score. ANIMALS: 13 horses with recurrent airway obstruction (RAO) and 6 control horses. PROCEDURE: 6 RAO affected and 6 control horses were stabled for 3 days. Coughing was counted for 4 hours before and on each day horses were stabled. Before and on day 3 of stabling, tracheal mucus accumulation was scored, airway obstruction was assessed via maximal change in pleural pressure (deltaPpl(max)), and airway inflammation was evaluated by use of cytologic examination of bronchoalveolar lavage fluid (BALF). Effects of dexamethasone (0.1 mg/kg, IV, q 24 h for 7 days) were determined in 12 RAO-affected horses. RESULTS: To assess frequency, coughing had to be counted for 1 hour. In RAO-affected horses, stabling was associated with increases in cough frequency, mucus score, and deltaPpl(max). Control horses coughed transiently when first stabled. In RAO-affected horses, coughing was correlated with deltaPpl(max), mucus score, and airway inflammation and was a sensitive and specific indicator of deltaPpl(max) > 6 cm H2O, mucus score > 1.0, and > 100 neutrophils/microL and > 20% neutrophils in BALF Dexamethasone reduced cough frequency, mucus score, and deltaPpl(max), but BALF neutrophil count remained increased. CONCLUSIONS AND CLINICAL RELEVANCE: Because of its sporadic nature, coughing cannot be assessed accurately by counting during brief periods. In RAO-affected horses, coughing is an indicator of airway inflammation and obstruction. Corticosteroid treatment reduces cough frequency concurrently with reductions in deltaPpl(max) and mucus accumulation in RAO-affected horses. PMID- 12755294 TI - Glomerular lesions in dogs infected with Leishmania organisms. AB - OBJECTIVE: To histologically identify glomerular lesions in dogs infected with Leishmania organisms. ANIMALS: 41 dogs (17 sexually intact males and 14 sexually intact and 10 ovariohysterectomized females) that had positive results when tested for leishmaniosis as determined by use of serologic evaluation (indirect fluorescent antibody test, titers of 1:80 to 1:640) and direct microscopic identification of the protozoal organisms. PROCEDURE: Urine samples were collected by use of cystocentesis and examined by qualitative SDS-agarose gel electrophoresis (AGE). All dogs had non-selective (glomerular) or mixed (glomerular and tubular) proteinemia. Specimens were obtained from each dog during ultrasound-assisted renal biopsy and used for histologic examination. Each specimen was stained with H&E, periodic acid-Schiff, Goldner's trichrome, methenamine silver, and Congo Red stains. Specimens were adequate for evaluation when they contained at least 5 glomeruli/section, except for specimens stained with Congo Red in which 1 glomerulus/section was adequate. RESULTS: Examination of renal biopsy specimens revealed various glomerular lesions in all dogs and interstitial or tubular (or both) lesions in 23 of 41 (55%) dogs. CONCLUSIONS AND CLINICAL RELEVANCE: Glomerular lesions that develop in dogs during infection with Leishmania organisms can be classified histologically as mesangial glomerulonephritis, membranous glomerulonephritis, membranoproliferative glomerulonephritis, and focal segmental glomerulonephritis. Tubulointerstitial histopathologic conditions were not observed as the primary lesion, despite being evident in 23 of 41 (55%) dogs. Use of SDS-AGE for qualitative evaluation of proteinuria and successive collection of specimens during renal biopsies following diagnosis of nonselective glomerular proteinuria provides the possibility for early identification of renal lesions. PMID- 12755295 TI - Measurement of glycated hemoglobin percentages for use in the diagnosis and monitoring of diabetes mellitus in nonhuman primates. AB - OBJECTIVE: To identify a technique for measurement of glycated hemoglobin percentage in blood samples obtained from various species of nonhuman primates (NHPs), to determine whether these percentages varied with respect to glycemic control, and to assess whether this physiologic variable provided a suitable test for diagnosing diabetes mellitus in NHPs. SAMPLE POPULATION: 166 blood samples collected from 121 NHPs comprising 22 species from the Haplorhine and Strepsirhine suborders and including nondiabetic, treated-diabetic, and diabetic animals in 23 zoologic institutions throughout the United States. PROCEDURE: Hemoglobin A1c percentage was measured in 154 samples by use of high-performance liquid chromatography. Total glycated hemoglobin percentage was measured in 159 samples by use of a boronate-affinity chromatographic assay. Glucose concentration was measured in 157 samples with an autochemical analyzer by use of a hexose kinase method. RESULTS: The boronate-affinity chromatographic technique for measurement of total glycated hemoglobin percentage was the most suitable method. Nondiabetic Haplorhines had percentages higher than those in nondiabetic Strepsirhines. In Haplorhines, diabetic animals had percentages higher than those in treated-diabetic animals, which had percentages higher than those in nondiabetic animals. In Strepsirhines, this pattern was less pronounced. CONCLUSIONS AND CLINICAL RELEVANCE: Measurement of total glycated hemoglobin percentage provides useful information for diagnosing diabetes mellitus in Haplorhines and, possibly, in Strepsirhines. Until reference ranges are established for each species, it is recommended that results for samples from NHPs without clinical signs of diabetes mellitus be compared with results of samples collected concomitantly from NHPs with clinical signs of this condition. PMID- 12755296 TI - Effect of bone diameter and eccentric loading on fatigue life of cortical screws used with interlocking nails. AB - OBJECTIVE: To test the effects of bone diameter and eccentric loading on fatigue life of 2.7-mm-diameter cortical bone screws used for locking a 6-mm-diameter interlocking nail. SAMPLE POPULATION: Eighteen 2.7-mm-diameter cortical bone screws. PROCEDURE: A simulated bone model with aluminum tubing and a 6-mm diameter interlocking nail was used to load screws in cyclic 3-point bending. Group 1 included 6 screws that were centrally loaded within 19-mm-diameter aluminum tubing. Group 2 included 6 screws that were centrally loaded within 31.8 mm-diameter aluminum tubing. Group 3 included 6 screws that were eccentrically loaded (5.5 mm from center) within 31.8-mm-diameter aluminum tubing. The number of cycles until screw failure and the mode of failure were recorded. RESULTS: An increase in the diameter of the aluminum tubing from 19 to 31.8 mm resulted in a significant decrease in the number of cycles to failure (mean +/- SD, 761,215 +/- 239,853 to 16,941 +/- 2,829 cycles, respectively). Within 31.8-mm tubing, the number of cycles of failure of eccentrically loaded screws (43,068 +/- 14,073 cycles) was significantly greater than that of centrally loaded screws (16,941 +/ 2,829 cycles). CONCLUSIONS AND CLINICAL RELEVANCE: Within a bone, locking screws are subjected to different loading conditions depending on location (diaphyseal vs metaphyseal). The fatigue life of a locking screw centrally loaded in the metaphyseal region of bone may be shorter than in the diaphysis. Eccentric loading of the locking screw in the metaphysis may help to improve its fatigue life. PMID- 12755297 TI - Validation and comparison of the use of diuresis cystometry and retrograde filling cystometry at various infusion rates in female Beagle dogs. AB - OBJECTIVES: To compare retrograde filling cystometry at infusion rates of 5, 10, and 20 mL/min with diuresis cystometry for determination of an appropriate infusion rate and to confirm the reproducibility of measurements obtained by urethral pressure profilometry (UPP) and cystometry in female Beagles. ANIMALS: Adult female Beagles. PROCEDURE: Successive UPP and cystometry were performed by use of a water perfusion catheter on dogs anesthetized with propofol. Dogs randomly underwent each of the following at 1-week intervals: retrograde filling cystometry at 5, 10, and 20 mL/min, and diuresis cystometry. The maximum urethral pressure and closure pressure, functional and anatomic profile lengths, threshold pressure, threshold volume, and compliance were measured. RESULTS: For each UPP variable, significant differences were found among dogs, but no significant differences were found in intra- or interstudy measurements for individual dogs. For retrograde filling cystometry, threshold pressure was not significantly different between a 5 and 10 mL/min infusion rate. Threshold pressure was significantly higher during retrograde filling cystometry at 20 mL/min, compared with 5 and 10 mL/min, and was associated with bladder wall damages. Threshold pressure was significantly lower during diuresis cystometry, compared with retrograde filling cystometries. Threshold volume and compliance were not significantly different among retrograde filling cystometries but were significantly higher during diuresis cystometry. CONCLUSIONS AND CLINICAL RELEVANCE: Retrograde filling cystometry at 20 mL/min leads to unacceptable sudden increase in threshold bladder pressure. Retrograde filling cystometry at 10 mL/min can be recommended in a clinical setting, shortening the anesthesia time. However, diuresis cystometry approximates physiologic bladder filling most accurately. PMID- 12755298 TI - Effect of intranasal exposure to leukotoxin-deficient Mannheimia haemolytica at the time of arrival at the feedyard on subsequent isolation of M haemolytica from nasal secretions of calves. AB - OBJECTIVE: To determine the effect of intranasal exposure to live leukotoxin (LktA)-deficient Mannheimia haemolytica (MH) at the time of feedyard arrival on nasopharyngeal colonization by wild-type MH in calves. ANIMALS: 200 calves. PROCEDURE: Calves from Arkansas (AR calves; n = 100; mean body weight, 205 kg) were purchased from an order buyer barn. Calves from New Mexico (NM calves; n = 100; mean body weight, 188 kg) were obtained from a single ranch. Calves were transported to a feedyard, where half of each group was exposed intranasally with LktA-deficient MH at the time of arrival. Calves were observed daily for respiratory tract disease (RTD), and nasal swab specimens were collected periodically to determine nasopharyngeal colonization status with MH. Serum samples were assayed for antibodies to MH. RESULTS: 15 AR calves had nasopharyngeal colonization by wild-type MH at the order buyer barn, whereas none of the NM calves had nasopharyngeal colonization. Intranasal exposure to LktA deficient MH elicited an increase in serum antibody titers against MH in NM calves, but titers were less in NM calves treated for RTD. Exposure of NM calves to LktA-deficient MH offered protection from nasopharyngeal colonization by wild type MH. CONCLUSIONS AND CLINICAL RELEVANCE: Exposure of calves to LktA-deficient MH elicited an increase in serum antibody titers against MH and decreased colonization of the nasopharynx by wild-type MH. Earlier exposure would likely allow an immune response to develop before transportation and offer protection from nasopharyngeal colonization and pneumonia caused by wild-type MH. PMID- 12755299 TI - Comparison of radiographic assessments of the tibial plateau slope in dogs. AB - OBJECTIVES: To evaluate the accuracy of 2 radiographic methods used to assess tibial plateau slope (TPS) in dogs and evaluate effects of film digitization and radiographic beam placement on TPS measurements. SAMPLE POPULATION: 16 hind limbs from dog cadavers weighing > 20 kg. PROCEDURES: Radiographs of tibiae were made with the radiographic beam centered over the stifle joint and midshaft of the tibia. Tibiae were collected, the femorotibial contact area was determined, and slope of the medial tibial condyle in relation to the tibial shaft was measured. Radiographs were digitized. Slope of the medial tibial condyle was measured on printed and digitized radiographs read in random order by 6 examiners unaware of anatomic measurements. Three examiners used a conventional measuring technique, and 3 examiners used an alternative measuring technique. RESULTS: Anatomic measurements were significantly higher than radiographic measurements made by use of the conventional interpretation method but did not differ from radiographic measurements made by use of the alternate method. Measurements from printed radiographs were lower than measurements from digitized radiographs for the 4 most experienced examiners. CONCLUSIONS AND CLINICAL RELEVANCE: Measurements made by use of a line tangential to the cranial, linear portion of the medial tibial condyle at the femorotibial contact point were accurate measurements of the anatomic TPS. Measurements made by use of the conventional TPS measurement method underestimated the anatomic TPS. Measurements made on digitized radiographs were typically more accurate than measurements made on printed radiographs. PMID- 12755300 TI - Effects of acepromazine on renal function in anesthetized dogs. AB - OBJECTIVE: To investigate the effects of IM administration of acepromazine on indices of relative renal blood flow and glomerular filtration rate (GFR) by means of scintigraphy, as well as the effects on physiologic, hematologic, and serum biochemical variables in anesthetized dogs, compared with effects of administration of saline. ANIMAL: 6 healthy Beagles. PROCEDURE: Acepromazine (0.1 mg/kg) or physiologic saline (0.9 NaCI) solution was administered IM 30 minutes prior to induction of anesthesia with thiopentone; anesthesia was maintained with inspired isoflurane for 2.25 hours. Blood gases and circulatory and ventilatory variables were monitored. Renal function was evaluated by scintigraphic measurements of GFR and relative renal blood flow and analyses of serum and urine. Statistical analyses used ANOVA or Friedman ANOVA. RESULTS: Values of relative renal blood flow and GFR remained high despite low blood pressures. After administration of acepromazine, mean +/- SD arterial blood pressure was 66 +/- 8 mm Hg during anesthesia; this value was below the threshold (80 mm Hg) for renal autoregulation of GFR. In comparison, mean arterial blood pressure after administration of saline was significantly higher (87 +/- 13 mm Hg). However, between treatments, there were no significant differences in GFR, relative renal blood flow, or other indices of renal function. CONCLUSIONS AND CLINICAL RELEVANCE: Measurements of renal function and blood flow in dogs during anesthesia with thiopentone and isoflurane did not differ significantly between treatments, which suggested that acepromazine protects renal function despite inducing reduction in blood pressure, compared with effects of administration of saline. PMID- 12755301 TI - Assessment of acid-base status and plasma lactate concentrations in arterial, mixed venous, and portal blood from dogs during experimental hepatic blood inflow occlusion. AB - OBJECTIVE: To determine the effects of extended experimental hepatic blood flow occlusion (ie, portal triad clamping [PTC]) in dogs by measuring acid-base status and plasma lactate concentrations in arterial, mixed venous, and portal blood and evaluating the relationship between metabolic and concurrent hemodynamic changes. ANIMALS: 6 healthy Beagles. PROCEDURE: During anesthesia with isoflurane, cardiac output and arterial blood pressure were measured. Arterial, mixed venous, and portal blood samples were collected simultaneously for blood gas analyses and plasma lactate measurements before PTC and at 8-minute intervals thereafter. RESULTS: PTC resulted in severe hemodynamic and metabolic alterations. Eight minutes after PTC, significant decreases in cardiac index from a baseline value of 3.40 +/- 0.27 to 1.54 +/- 0.26 L/min/m2 and in mean arterial blood pressure from a baseline value of 74 +/- 6 to 43 +/- 6 mm Hg were recorded. After PTC, results indicative of lactic acidosis were found in portal blood at 16 minutes, in mixed venous at 32 minutes, and in arterial blood at 48 minutes. Significant differences in measured variables were also found between arterial and portal blood samples, between mixed venous and portal blood samples, and between arterial and mixed venous blood samples after PTC, compared with differences at baseline. CONCLUSIONS AND CLINICAL RELEVANCE: Analysis of mixed venous blood is preferable to analysis of arterial blood in the assessment of metabolic derangement. In a clinical setting, occluded portal blood is released to the systemic circulation, and the degree of reperfusion injury may depend on the metabolic status of pooled portal blood. PMID- 12755302 TI - Two-dimensional link-segment model of the forelimb of dogs at a walk. AB - OBJECTIVE: To calculate normative joint angle, intersegmental forces, moment of force, and mechanical power at elbow, antebrachiocarpal, and metacarpophalangeal joints of dogs at a walk. ANIMALS: 6 clinically normal mixed-breed dogs. PROCEDURE: Kinetic data were collected via a force platform, and kinematic data were collected from forelimbs by use of 3-dimensional videography. Length, location of the center of mass, total mass, and mass moment of inertia about the center of mass were determined for each of 4 segments of the forelimb. Kinematic data and inertial properties were combined with vertical and craniocaudal ground reaction forces to calculate sagittal plane forces and moments across joints of interest throughout stance phase. Mechanical power was calculated as the product of net joint moment and the angular velocity. Joint angles were calculated directly from kinematic data. RESULTS: All joint intersegmental forces were similar to ground reaction forces, with a decrease in magnitude the more proximal the location of each joint. Flexor moments were observed at metacarpophalangeal and antebrachiocarpal joints, and extensor moments were observed at elbow and shoulder joints, which provided a net extensor support moment for the forelimb. Typical profiles of work existed for each joint. CONCLUSIONS AND CLINICAL RELEVANCE: For clinically normal dogs of a similar size at a walk, inverse dynamic calculation of intersegmental forces, moments of force, and mechanical power for forelimb joints yielded values of consistent patterns and magnitudes. These values may be used for comparison in evaluations of gait in other studies and in treatment of dogs with forelimb musculoskeletal disease. PMID- 12755303 TI - Dimensions and histologic characteristics of the small intestine of dogs during postnatal development. AB - OBJECTIVE: To quantify dimensions of the small intestine of dogs and describe changes in histologic characteristics of the mucosa during postnatal development. SAMPLE POPULATION: Gastrointestinal tract tissues obtained from 110 Beagles (15 adult females and 95 puppies of both sexes). PROCEDURE: Several variables (length, total weight, mucosal weight, and nominal surface area) of the small intestine were measured in puppies at birth but before suckling; 1 day after birth and subsequent suckling, 21, 42, and 63 days after birth, and in the adult dams of the puppies. Tissue structure was examined and quantified at each time point by use of routine histologic examination and ocular micrometry of formalin fixed specimens stained with H&E. RESULTS: Small intestinal dimensions increased throughout development with the greatest proportional changes during the first day after birth and onset of suckling. Villus height decreased during suckling but had consistent values from 42 days after birth to maturity, whereas crypt depth increased from birth to maturity. Vacuolated enterocytes were evident from birth to 21 days but not thereafter. CONCLUSIONS AND CLINICAL RELEVANCE: Increases in intestinal dimensions provide growing dogs with a greater capacity for digestion and absorption. Changes in mucosal architecture and cell populations coincided with shifts in dietary inputs. These findings may assist in the diagnosis of small intestinal diseases and nutritional responses during growth and development of dogs. PMID- 12755305 TI - Postnatal development of nutrient transport in the intestine of dogs. AB - OBJECTIVE: To measure nutrient absorption by the intestine during postnatal development of dogs. ANIMAL: 110 Beagles ranging from neonatal to adult dogs. PROCEDURE: Rates of absorption for sugars (glucose, galactose, and fructose), amino acids (aspartate, leucine, lysine, methionine, and proline), a dipeptide (glycyl-sarcosine), and linoleic acid by the proximal, mid, and distal regions of the small intestine were measured as functions of age and concentration (kinetics) by use of intact tissues and brush-border membrane vesicles. Absorption of octanoic acid by the proximal portion of the colon was measured in intact tissues. RESULTS: Rates of carrier-mediated transport by intact tissues decreased from birth to adulthood for aldohexoses and most amino acids but not for fructose and aspartate. Kinetics and characteristics of absorption suggest that there were changes in the densities, types, and proportions of various carriers for sugars and amino acids. Saturable absorption of linoleic acid in the small intestine and octanoic acid in the proximal portion of the colon increased after weaning. CONCLUSIONS AND CLINICAL RELEVANCE: Rates of absorption decreased between birth and adulthood for most nutrients. However, because of intestinal growth, absorption capacities of the entire small intestine remained constant for leucine and proline and increased for glucose, galactose, fructose, aspartate, and proline but were less than predicted from the increase in body weight. Although postnatal ontogeny of nutrient absorption was consistent with changes in the composition of the natural and commercial diets of growing dogs, rates of amino acid and peptide absorption were lower than expected. PMID- 12755304 TI - Activities of gastric, pancreatic, and intestinal brush-border membrane enzymes during postnatal development of dogs. AB - OBJECTIVE: To measure activities of digestive enzymes during postnatal development in dogs. SAMPLE POPULATION: Gastrointestinal tract tissues obtained from 110 Beagles ranging from neonatal to adult dogs. PROCEDURE: Pepsin and lipase activities were measured in gastric contents, and amylase, lipase, trypsin, and chymotrypsin activities were measured in small intestinal contents and pancreatic tissue. Activities of lactase, sucrase, 4 peptidases, and enteropeptidase were assayed in samples of mucosa obtained from 3 regions of the small intestine. RESULTS: Gastric pH was low at all ages. Pepsin was not detected until day 21, and activity increased between day 63 and adulthood. Activities of amylase and lipase in contents of the small intestine and pancreatic tissue were lower during suckling than after weaning. Activities of trypsin and chymotrypsin did not vary among ages for luminal contents, whereas activities associated with pancreatic tissue decreased between birth and adulthood for trypsin but increased for chymotrypsin. Lactase and gamma-glutamyltranspeptidase activities were highest at birth, whereas the activities of sucrase and the 4 peptidases increased after birth. Enteropeptidase was detected only in the proximal region of the small intestine at all ages. CONCLUSIONS AND CLINICAL RELEVANCE: Secretions in the gastrointestinal tract proximal to the duodenum, enzymes in milk, and other digestive mechanisms compensate for low luminal activities of pancreatic enzymes during the perinatal period. Postnatal changes in digestive secretions influence nutrient availability, concentrations of signaling molecules, and activity of antimicrobial compounds that inhibit pathogens. Matching sources of nutrients to digestive abilities will improve the health of dogs during development. PMID- 12755306 TI - Postnatal changes in bacterial populations in the gastrointestinal tract of dogs. AB - OBJECTIVE: To describe postnatal changes in the populations of bacteria in the gastrointestinal tract (GIT) of dogs. ANIMALS: 110 Beagles ranging from neonatal to adult dogs. PROCEDURE: Contents of the stomach and proximal and distal portions of the colon and contents and mucosa of the mid region of the small intestine were collected from puppies at 1 day after birth and subsequent suckling; puppies at 21, 42, and 63 days after birth; and adult female dogs (ie, dams of the puppies) for enumeration of bacterial populations. RESULTS: The entire GIT was colonized at day 1 by all groups of bacteria studied; aerotolerant forms were dominant. During subsequent postnatal development, there were changes in the relative proportions of the various groups of bacteria with anaerobic groups increasing in absolute and relative numbers. CONCLUSIONS AND CLINICAL RELEVANCE: Establishment of bacterial populations in the GIT of dogs is a gradual process that begins immediately after birth. Age-related changes in the relative proportions of bacterial groups coincided with changes in diet and physiologic processes of the host and can influence nutritional state and disease resistance of developing dogs. Differences among regions of the GIT suggest that fecal samples may have limited use for understanding the populations of bacteria and the age and diet-related changes in various regions of the GIT. PMID- 12755307 TI - The history of tracheal surgery. AB - Tracheal surgery, which did not exist in a coherent, systematic fashion 45 years ago, has developed techniques that allow resection of approximately half of the adult trachea with primary reconstruction, largely by anatomic mobilization procedures. Dependable methods have also been developed for laryngotracheal and carinal resection and reconstruction. The daunting problem of long congenital tracheal stenosis appears to be largely solved by slide tracheoplasty. In the past four decades much has also been learned about the etiology, natural history, pathology, and (in some cases) prevention of tracheal diseases including primary and secondary tumors, postintubation injuries, and idiopathic stenosis. PMID- 12755308 TI - Surgical anatomy of the trachea. AB - A clear understanding of tracheal anatomy is a fundamental building block for surgical procedures. This article describes the surgical anatomy of the trachea and emphasizes how these anatomic facts affect surgical procedures performed on the trachea. PMID- 12755309 TI - Tracheal release maneuvers. AB - Careful preoperative trachea "mapping" with accurate measurements should make an unexpected shortfall in remaining tracheal length and mobility uncommon. Bronchoscopy is the most accurate means of "mapping" the pathologic tracheal anatomy. Rigid bronchoscopy using the ventilating bronchoscope is preferred over flexible bronchoscopy because it provides the most accurate means of measuring the length of stenotic trachea (especially when used in conjunction with rigid fiberoptic telescopes), it can be used to dilate a tracheal stenosis before operative repair, and it establishes a safe airway under direct vision. When the anatomy of the tracheal pathology is accurately defined, specific plans for tracheal resection can be established to accomplish a well-vascularized, low tension tracheal anastomosis. PMID- 12755310 TI - Postintubation tracheal stenosis. AB - Postintubation tracheal stenosis is a clinical problem caused by regional ischemic necrosis of the airway. The incidence of postintubation tracheal stenosis has decreased with recognition of its etiology and modifications in the design and management endotracheal and tracheostomy tubes; however, it remains the most common indication for tracheal resection and reconstruction. Single stage resection and reconstruction by a competent tracheal surgeon results in good or satisfactory results in 93.7% of patients, with a failure rate of 3.9% and a mortality rate of 2.4%. The intellect and skill of Dr. Grillo has made the etiology and management of postintubation stenosis obvious to us all. PMID- 12755311 TI - Primary tracheal tumors. AB - Primary tracheal tumors often present with locally advanced tumors. A majority of patients can safely undergo tracheal, laryngotracheal, or carinal resection with low perioperative risk. Airway interventions at the time of diagnosis should be selected carefully to avoid a compromise of curative treatment. Precise judgment is required to determine resectability. The proximity of intrathoracic organs creates anatomical limits to en bloc resection and necessitates adjuvant radiotherapy in malignant tumors. Early referral for consideration of surgical resection might offer the best opportunity for improving the overall prognosis of tracheal tumors. PMID- 12755312 TI - Idiopathic laryngotracheal stenosis. AB - ILTS is a rare inflammatory disease that results in a cicatricial stenosis of the cricoid and upper trachea. It occurs almost exclusively in women and is without known cause. Patients present with dyspnea on exertion that progresses to dyspnea at rest, often with stridor. The diagnosis is usually made on the basis of patient history, physical examination, and radiography. Rigid bronchoscopy is usually reserved for the day of proposed surgery to confirm diagnosis and to plan the operative strategy. On occasion, the presence of active inflammation extending into the immediate subglottis or the patient's use of corticosteroids requires that surgery be postponed. In these cases, a patent airway is temporarily restored with careful bronchoscopic dilation. Single-staged laryngotracheal resection is successful in more than 90% of patients and is the most effective treatment when performed by experienced hands. Long-term follow-up shows stable airway and improvement in voice quality. Palliative procedures such as repeated airway dilations should be reserved for poor surgical candidates. Protective tracheostomy is rarely required. PMID- 12755313 TI - Tracheoesophageal fistula. AB - Acquired TEF is a rare complication that can occur from a variety of causes. The most common etiology of nonmalignant TEF is as a complication of intubation with cuff-related tracheal injury. Most patients present with increased secretions, pneumonia, and evidence of aspiration of gastric contents while the patient is on mechanical ventilation. When diagnosed after extubation, the most frequent sign of TEF is coughing after swallowing. A high index of suspicion is required in patients at risk for developing a TEF. The diagnostic evaluation is by bronchoscopy and esophagoscopy. When the diagnosis has been made, the immediate goal should be to minimize tracheobronchial soilage by placing the cuff of a tracheostomy tube distal to the fistula. Reflux of gastric contents is diminished by placement of a gastrostomy tube, and adequate nutrition is facilitated by inserting a jejunostomy tube. Surgical correction is required because spontaneous closure is rare, but surgery should be postponed until the patient is weaned from mechanical ventilation because positive pressure ventilation after tracheal repair carries an increased risk of anastomotic dehiscence and restenosis. An anterior cervical collar incision can be used for most cases of post-intubation TEFs. The esophagus should be closed in two layers over a nasogastric tube and buttressed with a pedicled strap muscle flap. If the tracheal defect is small, primary repair can be employed. In most cases, however, the best results can be achieved with tracheal resection and reconstruction. The patient should be extubated at the completion of the case, if possible. With this strategy, as first described by Grillo and colleagues [27], single-stage repair can be performed safely and with a high success rate. Malignant TEFs cannot be cured because of the underlying incurable disease process. As with nonmalignant TEFs, the principal complications are tracheo-bronchial contamination and poor nutrition. Without prompt palliation, death occurs rapidly, with a mean survival time of between 1 and 6 weeks in patients who are treated with supportive care alone. The most common primary tumor causing malignant TEF is esophageal carcinoma. The other frequent cause is lung cancer. Patients present with signs and symptoms typical of TEF, including coughing after swallowing. Diagnosis is made by barium esophagography, and the location and size of the fistula is determined by bronchoscopy and esophagoscopy. Treatment must correct the two problems of airway contamination and poor nutrition. The most effective treatments are esophageal bypass and esophageal stenting. Bypass is demonstrated to resolve respiratory soilage and allow fairly normal swallowing, but it should be reserved for patients who can tolerate a major operation. Stenting can be offered to nearly all patients regardless of their physiologic condition. Stenting also limits aspiration and allows swallowing. Esophageal exclusion is rarely indicated in the current era of familiarity with stenting techniques. Direct fistula closure and fistula resection do not yield satisfactory results. Radiation therapy and chemotherapy combined might offer a survival benefit compared with supportive care alone. The complication of TEF secondary to malignancy is a devastating problem that carries a bleak prognosis, but when it is performed promptly after the diagnosis of a malignant TEF, esophageal bypass or stenting improves survival and quality of life for these unfortunate patients. PMID- 12755314 TI - Tracheal trauma. AB - The etiology, presentation, and management of blunt and penetrating injuries of the trachea has been reviewed. The approach to and outcome following management of more unusual situations such as iatrogenic injuries has also been briefly reviewed. Early recognition of these problems and careful attention to the details of acute management can convert a life-threatening situation into one that can usually be successfully managed by the techniques of tracheal surgery developed and popularized by Dr. Grillo. PMID- 12755315 TI - Pediatric tracheal surgery. AB - Pediatric tracheal surgery is uncommon, and few centers have enough experience to make meaningful conclusions about treatment. Short-segment congenital tracheal stenosis is treated by tracheal resection, whereas long-segment stenosis is treated by either augmentation tracheoplasty or slide tracheoplasty (the author's preferred approach). Tracheomalacia is treated most commonly by aortopexy. Postintubation tracheal stenosis is usually treated by tracheal (or laryngotracheal) resection. PMID- 12755316 TI - Carinal resection and reconstruction. AB - Successful outcomes for carinal resection and reconstruction depend upon many factors. Careful patient selection for patients who can tolerate the physiologic effects of the operation cannot be underestimated. Understanding the safe limits of resection, the technical details of airway reconstruction, and ongoing improvements in intraoperative and postoperative care should minimize the morbidity and mortality rates previously reported with this procedure. In addition, further work from institutions with considerable experience in carinal resection is needed to better define the long-term outcome for patients with bronchogenic carcinoma in close proximity to or involving the carina. Prior studies have suggested reasonable survival rates can be expected in the absence of involved mediastinal nodes or distant metastatic disease. PMID- 12755317 TI - Tracheoinnominate fistula: diagnosis and management. AB - TIF is a rare and often fatal complication of tracheostomy. Bleeding from the trachea after tracheostomy demands urgent investigation. Bronchoscopy is the diagnostic procedure of choice. Bedside control of hemorrhage by cuff overinflation or by digital arterial compression can be lifesaving. Prompt operation with division of the innominate artery and subsequent separation of the trachea from the divided artery by viable tissue is indicated. Neurologic complications are rare. PMID- 12755318 TI - The burned trachea. AB - Tracheal burns caused by thermal or chemical injuries are uncommon, but they can be difficult to manage. A high index of suspicion is required for early detection of these injuries. Acutely, they are often life threatening unless they are diagnosed quickly and managed properly. Tracheal burn injuries can lead to long term stenoses, which require specialized surgical techniques and chronic therapy- often with Montgomery T-tubes--for their proper management. PMID- 12755319 TI - Tracheomalacia. AB - Tracheomalacia is a rare tracheal problem that leads to collapse of the airway and expiratory flow obstruction. Expiratory CT scans are the diagnostic test of choice in adults. Chronic obstructive pulmonary disease is the most common cause of adult tracheomalacia. Plication of the membranous wall to polypropylene mesh to recreate the normal airway shape (membranous wall tracheoplasty) is an effective treatment in adults. PMID- 12755320 TI - Surgical management of thyroid carcinoma invading the trachea. AB - Airway invasion by thyroid carcinoma is an uncommon but important clinical problem. The surgical management of airway invasion is somewhat controversial, with some studies suggesting that conservative shave procedures might be adequate; however, the standardization and safety of techniques of airway resection and reconstruction have made en bloc surgery a reasonable approach for the management of such carcinomas. Tracheal resection and reconstruction for thyroid carcinomas with airway invasion might provide long-lasting palliation and might even be curative in a significant number of patients suffering from this disease. PMID- 12755321 TI - Management of persistent tracheal stoma. AB - A rare late complication of tracheostomy is persistence of the stoma after removal of the tracheostomy tube. This article reviews the indications for surgical repair and outlines two alternative approaches to surgical management of this problem. PMID- 12755322 TI - Reoperative tracheal surgery. AB - Tracheal resection and reconstruction for postintubation stenosis is successful in more than 95% of initial repair attempts. The most likely causes of anastomotic failure are anastomotic tension, local devascularization, and granulomatous foreign body reaction. Incomplete resection of areas of stenosis or malacia might also lead to postoperative airway compromise. A variety of systemic factors might contribute to poor anastomotic healing. Postoperative respiratory difficulty requires immediate evaluation. In a patient with recurrent tracheal stenosis, the airway can be managed with dilation, or a tracheostomy or T-tube can be inserted through the failed anastomosis. Patients who are candidates for reoperative tracheal resection and reconstruction can expect good or satisfactory results in 91.9% of cases. Preoperatively addressing the patient's risk factors for failing, and liberally employing release procedures to reduce tension on the anastomosis contribute to the success of a reoperative procedure. PMID- 12755323 TI - Management of complications of tracheal surgery. AB - Basic principles of tracheal reconstruction, which were introduced in the 1960s and 1970s, served to reduce the prevalence of many complications. These principles include thorough preoperative assessment (endoscopic and radiologic) of the tracheal anatomy and glottic function, avoidance of excessive anastomotic tension, preservation of tracheal blood supply, and meticulous dissection and anastomosis. The tracheal surgeon should have access to expert help in radiology and anesthesiology, experienced nursing units, and the help and advice of consultants, especially otolaryngologists. The surgical approach should be meticulously planned. No irreversible maneuvers should be performed until one establishes certainty to proceed to resection. The surgeon should not attempt to exceed the limits of what appears to be reasonably possible. It must be remembered that a permanent tracheal T-tube might be the best solution for a patient with extensive tracheal damage that would defy reconstruction. PMID- 12755324 TI - Personality disorder and Axis I psychopathology: the problematic boundary of Axis I and Axis II. AB - The confusion of personality disorders with Axis I disorders can be traced in part to inadequacies of assessment instruments and diagnostic criterion sets. However, it also reflects the absence of adequate conceptualization. If Axis I continues to include early onset, chronic impairments that characterize everyday functioning, then there is unlikely to be a clear or meaningful distinction. Inherent and unique to personality disorders is that they concern a person's sense of self and identity. They are disorders of everyday functioning. Personality disorders have an early onset, characterize everyday functioning, and relate closely to personality functioning evident within the general population; Axis I disorders, in contrast, have an onset throughout adult life, are episodic, and are readily distinguishable from normal personality functioning. PMID- 12755325 TI - Personality and psychopathology: working toward the bigger picture. AB - There are systematic and meaningful links among normal and abnormal personality traits and Axis I and II constructs from the DSM. Nevertheless, much research in this area focuses on pairs of constructs (e.g., the link between personality traits and a specific Axis I disorder), rather than on the broader multivariate structure of the personality-psychopathology domain. We underscore the need for this broader perspective, a perspective that would transcend largely artificial boundaries between current constructs (e.g., normal and abnormal personality). We outline our approach to research from this perspective and we emphasize the internalizing (mood and anxiety) and externalizing (substance use and antisocial behavior) spectra as promising foci for initial research on the joint structure of personality and psychopathology. PMID- 12755326 TI - The core elements of neurosis: mixed anxiety-depression (cothymia) and personality disorder. AB - Although there has been great diagnostic activity within the conditions formally included under the general rubric of neurosis in the last 20 years, there is little evidence that the many new diagnoses (i.e., generalized anxiety disorder, panic disorder, social anxiety disorder, and dysthymic disorder) have helped clinicians and improved the health of those diagnosed. This is largely because of the extensive comorbidity between these disorders negates much of their attempted separation and it is argued that the core of neurosis is a mixed anxiety depressive disorder, or cothymia, combined with significant personality disorder of any type. The specific association of the anxious-fearful personality cluster (cluster C) and neurosis, called the general neurotic syndrome, is also relevant but appears to have lesser significance as the personality elements are not stable. Data are presented that justify these conclusions from a long-term follow up study of anxiety and depressive disorders. PMID- 12755327 TI - Personality and anxiety disorders: a review. AB - In this article, we summarize research on how normal personality and personality disorder traits may relate to anxiety disorders as predisposing factors, complications, and results of common underlying etiologies. We outline important questions and how these may be addressed through future research using genetically informative longitudinal and other designs, including: Are high neuroticism/cluster C personality traits causally related to the development of anxiety disorders? To what extent does the state of having an anxiety disorder influence the assessment of personality traits? Do high neuroticism/personality disorder traits and anxiety disorders co-occur because of shared genetic and environmental determinants? And, do personality disorder traits add to the prediction of anxiety disorders when normal personality traits are taken into account? PMID- 12755328 TI - Separate personality traits from states to predict depression. AB - Results have been inconsistent regarding the ability of personality measures to predict future depression severity levels, leading some researchers to question the validity of personality assessment, especially when patients are acutely depressed. Using a combination of regression and factor analytic techniques, we separated the variance of personality measures into stable trait and variable state-affect components. Findings supported the hypotheses that depression severity measured at different time points would correlate with both stable trait and concurrent state-affect components in personality measures, whereas change in depression severity would correlate with state changes but not with stable trait scores. Thus, personality assessments tap both state affect and trait variance, with the state-affect variance masking the trait variance when patients are depressed. PMID- 12755329 TI - Flamingo, a cadherin-type receptor involved in the Drosophila planar polarity pathway, can block signaling via the canonical wnt pathway in Xenopus laevis. AB - The Flamingo gene encodes a seven-pass transmembrane receptor of the cadherin super family and is one of a growing number of components identified as being necessary for the establishment of planar polarity in the Drosophila wing. Although vertebrate homologues of Flamingo have been identified in both man and mice, no function has as yet been ascribed to them. Here, we report the cloning of the Xenopus homologue of Flamingo (XFmi). XFmi is expressed in the dorsal ectoderm during gastrulation and in the forebrain and midbrain subsequently. We show that ectopic expression of the murine Flamingo gene can prevent the wnt mediated posteriorisation of the neural plate by interfering with the canonical wnt signalling pathway. PMID- 12755330 TI - Isolation and growth factor inducibility of the Xenopus laevis Lmx1b gene. AB - This paper reports the cloning of the full length Xenopus laevis Lmx1b gene, Xlmx1b. Xlmx1b is a LIM homeodomain protein with high conservation to homologues identified in human, mouse, hamster and chick. In situ hybridisation and RT-PCR analysis showed that Xlmx1b has a specific temporal expression pattern which can be separated into three main spatial domains. An Xlmx1b probe hybridized to regions of the nervous system from stage 13 onwards; these regions included the placodes and otic vesicles, the eye and specific sets of neurons. Sectioning of in situ hybridised embryos confirmed the location of transcripts as discreet regions of staining in ventrolateral regions of the neural tube. From stage 27, transcripts could be detected in the capsule of pronephric glomus. Finally, transcripts were detected by Northern blot analysis in the developing fore and hind limbs. Xlmx1b transcripts were also detected by Northern blot analysis in eye, brain, muscle and mesonephros tissue in metamorphosing tadpoles. RT-PCR analysis showed that zygotic expression of Xlmx1b is initiated at stage 10.5 and the temporal sequence of Xlmx1b expression is identical in both neural and presumptive pronephros regions. The effects of the growth factors activin A, retinoic acid (RA) and basic fibroblast growth factor (bFGF) on the regulation of Xlmx1b were also studied. Xlmx1b was found to be upregulated by activin A and RA inhibited this upregulation in a concentration dependant manner. In contrast, bFGF had no effect on the regulation of Xlmx1b. PMID- 12755331 TI - Notch activity is required to maintain floorplate identity and to control neurogenesis in the chick hindbrain and spinal cord. AB - Notch signalling plays a major role in many invertebrate and vertebrate patterning systems. In this paper, we use high-titre, non-replicative pseudotype viruses to show that the two Notch ligands, Delta1 and Serrate1 (Jagged1), have differing activities in the developing chick spinal cord and hindbrain. In the walls of the neural tube, Serrate1 appears not to affect neurogenesis, in contrast to Delta1 which mediates lateral inhibition as elsewhere in the nervous system. In the floorplate we find that there is also a requirement for Notch, but with a different type of dependence on the two Notch ligands: cells with a floorplate character are lost when Notch activity is blocked with dominant negative, truncated forms of either Delta1 or Serrate1. Our results are consistent with ligand-receptor specificity within the Notch signalling pathway, Serrate1 recognising selectively Notch2 (which is expressed in the floorplate), and Delta1 acting on both Notch2 and Notch1 (which is expressed in the walls of the neural tube). PMID- 12755332 TI - Multilineage hematopoietic progenitor activity generated autonomously in the mouse yolk sac: analysis using angiogenesis-defective embryos. AB - The capacity of the yolk sac to generate multilineage, adult-type hematopoiesis was investigated in vivo using vascular endothelial-cadherin deficient embryos. In these mutants, the yolk sac is not connected to the vasculature of the embryo and therefore all hematopoietic activity detected therein is intrinsic to the yolk sac and not derived from intraembryonic sources. At embryonic days 9.5 and 10.5, the yolk sac contains blood cells from the first wave of hematopoiesis, i.e. primitive erythrocytes and monocytes, but also multipotent progenitors from definitive hematopoiesis and a few granulocytes. Reverse transcription-polymerase chain reaction analysis revealed expression of specific genes of all lineages except lymphoid cells. Moreover, hematopoietic colony assays showed the existence of committed progenitors of the second wave of embryonic hematopoiesis, namely for definitive erythrocytes, megakaryocytes, granulocytes and monocytes. Conversely, the number of lymphocytes after lymphoid culture was insignificant. Our data provide evidence for multilineage hematopoiesis (but not lymphopoiesis) in the yolk sac in the absence of seeding from the embryo. The small number of definitive mature blood cells indicates however that the yolk sac is not an effective environment for the terminal differentiation of committed progenitors from the second wave of hematopoiesis. PMID- 12755333 TI - Analysis of the odontogenic and osteogenic potentials of dental pulp in vivo using a Col1a1-2.3-GFP transgene. AB - Recently, transgenic mice that carry a Green Fluorescent Protein (GFP) reporter gene fused to 2.3 kb fragment of rat Col1a1 regulatory sequences (pOBCol2.3GFPemd) were generated. In the present study, we have examined the patterns of expression of Col1a1-2.3-GFP during odontoblast differentiation in this transgenic line. We report that Col1a1-2.3-GFP is expressed in newly differentiated odontoblasts secreting predentin and fully differentiated odontoblasts. The pattern of expression of Col1a1-2.3-GFP in odontoblasts is correlated with that of dentin sialophosphoprotein (DSPP). Col1a1-2.3-GFP is also expressed in the osteoblasts and osteocytes of alveolar bone. The pattern of expression of Col1a1-2.3-GFP in osteocytes is correlated with the expression of Dmp1. These observations indicate the 2.3 kb rat Col1a1 promoter fragment has sufficient strength and specificity to monitor the stage-specific changes during both odontoblast and osteoblast differentiation. We also used coronal pulp tissues isolated from postnatal pOBCol2.3GFPemd transgenic animals to follow their differentiation after transplantation under the kidney capsule. Our observations provide direct evidence that the dental pulp contains competent progenitor cells capable of differentiating into new generations of odontoblast like cells which express high levels of Col1a1-2.3-GFP and DSPP and secrete tubular containing reparative dentin. We also report that the dental pulp is capable of giving rise to atubular bone-like tissue containing osteocytes expressing high levels of Col1a1-2.3-GFP and Dmp1. Our studies indicate that pOBCol2.3GFPemd transgenic animals provide a powerful tool for direct examination of the underlying mechanisms and the signaling pathways involved in dentin regeneration and repair, stem cell properties and heterogeneity of the dental pulp. PMID- 12755334 TI - Expression of DjXnp, a novel member of the SNF2-like ATP-dependent chromatin remodelling genes, in intact and regenerating planarians. AB - SWI/SNF-related complexes include proteins implicated in the regulation of gene expression by chromatin remodelling. We have identified in planarians, invertebrates well-known for their regenerative capability,the cDNA of a novel gene, DjXnp, which encodes a protein of 1,076 amino acids, containing seven helicase domains similar to those found in the SNF2-like family members. Sequence comparison reveals a significant degree of similarity of DjXNP with mammalian XNP/ATRX proteins. In situ hybridization experiments performed on intact and regenerating planarians demonstrated that DjXnp transcripts were distributed in mesenchymal cells and were especially abundant in nerve cells. During anterior regeneration, DjXnp was detected in the blastemal area where the nervous system is newly forming. This expression pattern reveals extensive similarities with that described for mammalian XNP/ATRX, suggesting that these genes may have a conserved function at the cellular level. PMID- 12755335 TI - Expression of muscle LIM protein during early development in Xenopus laevis. AB - We have isolated the Xenopus homologue of Muscle LIM protein (MLP, CRP3) and examined its expression during early embryonic development. MLP is only expressed in the differentiated heart during early development and is expressed in a subset of other striated muscles during later stages. There is no MLP expression during primary myogenesis in the somites, although it is found in adult skeletal muscle. PMID- 12755336 TI - Myogenic potential of mouse primordial germ cells. AB - Primordial germ cells are the only stem cells that retain true developmental totipotency after gastrulation, express markers typical of totipotent/pluripotent status and are able both in vivo and in vitro to give rise to pluripotent stem cells as EC and EG cells. We have therefore explored the possibility of the trans differentiation of mouse PGCs to a myogenic lineage by transplanting them directly or after in vitro culture into a regenerating muscle and by culturing them on monolayers of differentianting muscle cells. The results obtained suggest that mouse PGCs may trans-differentiate into myogenic cells, provided that their somatic environment is preserved. This occurs at an estimated frequency of 0.01%, which is no higher than that reported for stem cells of adult tissues. PMID- 12755337 TI - Developmental regulation of expression of Ran/M1 and Ran/M2 isoforms of Ran GTPase in mouse testis. AB - Two isoforms of Ran-GTPase have been described: Ran/M1 and Ran/M2. Ran/M2 is testis specific, whereas the Ran/M1 isoform is also expressed in somatic tissues. Here we show that both mRNAs, differing in 35 of the 648 nucleotides included in the ORFs, are developmentally regulated during spermatogenesis. Real-time RT-PCR experiments demonstrated that the expression of Ran/M1 and Ran/M2 increased in pachytene spermatocytes with progressive transcript accumulation until they reached the round spermatid stage, in the seminiferous epithelium of adults. In the testis, the expression of both isoforms was found to be restricted to germ cells. An expression window from early pachytene spermatocytes to late round spermatids was detected by in situ hybridization. PMID- 12755338 TI - Glycosaminoglycans in early chick embryo. AB - The developmental profile of glycosaminoglycans (GAGs) were examined by cellulose acetate electrophoresis and high performance liquid chromatography in the early chick embryo from late blastula (stage XIII+) to early somite developmental stages (stage HH7-9). Sulphated GAGs were present from the earliest stages. They were more abundant than the non-sulphated forms and showed stage-related changes. Chondroitin sulphate and especially dermatan sulphate appeared to be the predominant GAGs in embryos at stage XIII+. Dermatan sulphate was about three times as abundant as chondroitin sulphate at stage XII+. In contrast, embryos at the definitive streak stage (stage HH4) produced about twice as much chondroitin sulphate as dermatan sulphate. At the head process stage (stage HH5), the level of chondroitin sulphate was reduced and its relative content in the embryo was about the same as dermatan sulphate. Levels of dermatan sulphate were more than five times those of heparan sulphate from stage XIII through to stage HH5 and three times more at stage HH7-9. The 4- and 6- sulphation of chondroitin sulphate increased 14- and 10-fold respectively, from stage XIII+ to stage HH 7-9. The sulphation pattern of chondroitin sulphate had a delta(di)-4S:delta(di)-6S molar ratio ranging from 4 to 8:1 and a delta(di)-4S:delta(di)-OS molar ratio ranging from 9 to 16:1 and was developmentally regulated. Thus, chondroitin sulphate in the early chick embryo was sulphated predominately in the 4-position in all stages studied. The presence of both 4- and 6-sulphated disaccharides in chondroitin sulphate indicated that both 4 and 6 sulfotransferases were active in the early embryo. Hyaluronate and sulphated GAG content increased markedly at gastrulation when the first major cellular migrations and tissue interactions begin. PMID- 12755339 TI - Mrs Howlett's box and certificate. PMID- 12755340 TI - Pharmacology of progestogens used in oral contraceptives: an historical review to contemporary prescribing. PMID- 12755341 TI - Evaluation of the clinical usefulness of isolation of fetal DNA from the maternal circulation. AB - OBJECTIVE: To assess the reliability of isolating free fetal DNA from maternal usefulness. DESIGN: Fetal DNA was isolated from plasma or serum that was either collected prospectively or from archived samples collected for the purposes of second trimester screening. METHODS: Prospective samples were collected from patients undergoing prenatal diagnostic procedures (n = 24). A second group of samples from Rhesus negative women (n = 28) were assayed in which blood had originally been collected for maternal triple serum screening. DNA was extracted from all samples and assayed for the presence of the beta-globin gene, sex determing region Y (SRY) gene and Rh gene. All DNA sample handling and extraction was carried out by a single operator, and polymerase chain reaction (PCR) was carried out using previously published PCR primers and appropriate controls. The accuracy of results was assessed relative to the karyotype in the case of the SRY gene or cord blood phenotype in the case of the Rh gene. RESULTS: The SRY PCR results were compared to fetal cell karyotypes obtained from invasive diagnostic testing, 21 out of the 24 samples were correctly 'sexed'. The RhD PCR results were compared to fetal cord blood samples at the time of delivery, and showed both false positive and false negative results. Two RhD negative babies were genotyped as RhD positive, despite repeat analysis. CONCLUSION: It is possible to isolate fetal DNA from maternal serum. It is a potentially clinically useful technique in our laboratory and can be used to detect male fetuses, and Rh negative fetuses. To be useful in clinical practice, it is necessary to safeguard against contamination at the time of sample handling, and to use the optimal range of primers available to cover the polymorphisms present within the RhD gene. Although not robust enough yet to be used with diagnostic certainty in our hands, immense improvements in technique, probes and real-time PCR equipment make this type of diagnosis a reality in the near future. PMID- 12755342 TI - Prospective ranking of the sonographic markers for aneuploidy: data of 2143 prenatal cytogenetic diagnoses referred for abnormalities on ultrasound. AB - OBJECTIVE: To design a scheme to rank sonographic anomalies as indicators of aneuploidy and record the distribution of data from 2143 prenatal amniotic fluid/chorionic villous sample diagnoses referred for karyotyping because of fetal anomalies detected with ultrasound. METHODS: In all cases the records of sonographic anomalies were obtained prior to karyotyping. A cascade of seven prospective categories of ultrasound anomalies was chosen and the data were included in the highest compatible sonography category. The categories were in descending order: (I) combined central nervous system (CNS)/cranial shape and cardiac anomalies (excluding spina bifida and anencephaly); (II) key anomaly present (exomphalos/ intrauterine growth restriction/duodenal atresia/cystic hygroma/fetal hydrops/talipes--with other multiple anomalies); (III) CNS +/- other abnormality (excluding choroid plexus cyst, spina bifida, anencephaly); (IVa) increased nuchal translucency--first trimester +/- other abnormality; (IVb) increased nuchal thickening--second trimester +/- other abnormality; (V) cardiac anomaly +/- other abnormality; (VI) other markers of aneuploidy (pyelectasis/two vessel cord/echogenic bowel/short femur); and (VII) other (mostly isolated) malformations. RESULTS: There were 412/2143 (19.2%) chromosome abnormalities detected in this sonographically abnormal group. Overall, the prevalence of aneuploidy significantly ranged from 51 to 3% according to the above I-VII ultrasound categories and from approximately 1-80% for individual ultrasound anomalies. Likelihood ratios were derived for many ultrasound anomalies for several aneuploidy groups: trisomies of 13; 18; and 21; 45,X and 45,X mosaics; triploidy; other autosomal duplications and/or deletions; and other (than 45,X) sex chromosomal aneuploidies. CONCLUSION: It is suggested this data could be used to assist pre-procedural counselling of patients after the ultrasound scan in tertiary referral centres for prenatal cytogenetic diagnosis. PMID- 12755343 TI - Non-obstetric surgery during gestation: risk factors for lower birthweight. AB - OBJECTIVE: To assess the risk for preterm birth and low birthweight for women undergoing non-obstetric surgery during gestation. DESIGN: Two perinatal tertiary care centres. POPULATION: Women undergoing non-obstetric surgery during gestation between January 1989 and June 1999. MATERIALS AND METHODS: A chart review was carried out. Cervical cerclages, procedures carried out under local anaesthesia or intravenous sedation, or carried out in combination with Caesarean delivery were excluded. MAIN OUTCOME MEASURES: Preterm birth (<37 weeks), birthweight. RESULTS: A total of 116 of 69 800 women (0.2%) underwent non-obstetric surgery, with 96 women delivering under our care. Procedures were more commonly carried out in the second trimester (53%), versus the first (23%) or third trimester (24%). Surgery in the second trimester resulted in the lowest rate of preterm birth (11%). The overall preterm birth rate was 21% (20/96), with 13 out of 20 (65%) occurring between 35 and 37 weeks. The mean interval from surgery to delivery was 18.7 weeks. Rates of preterm birth were similar for either intra- versus extra-abdominal procedures, or general versus regional anaesthetic. Use of a general anaesthetic was associated with a significant decrease in birthweight (3053 vs 3515 g, P = 0.01) despite similar gestational ages at delivery (37.6 vs 38.6 weeks, P = 0.08). Multiple linear regression controlled for gestational age showed that general anaesthesia, longer surgery duration, and intra-abdominal procedures were all significant independent risk factors for lower birthweight. CONCLUSION: While non-obstetric surgery appears to be relatively safe during gestation, general anaesthesia, longer surgery time, and intra-abdominal procedures are associated with lower birthweights. PMID- 12755344 TI - Delivery of singleton preterm infants in New South Wales, 1990-1997. AB - OBJECTIVE: To examine trends in the maternal characteristics and delivery of singleton preterm infants in an Australian population. DESIGN: Population-based descriptive study. SETTING: New South Wales (NSW), Australia. POPULATION: The population included 37 500 singleton preterm births from 1 January 1990 to 31 December 1997. METHODS: Data were obtained from the NSW Midwives' Data Collection (MDC) and rates over time were calculated. Preterm birth by Caesarean section before the onset of labour or where labour was induced were considered to be medically indicated. MAIN OUTCOME MEASURES: Preterm rates, medically indicated preterm birth rates, mode of delivery andneonatal outcomes, and trends over time. RESULTS: Among singleton infants, there was no significant change over time in the rate of preterm birth (annual average 5.5%), preterm births that were medically indicated (annual average 29.3%) or neonatal outcomes of preterm births. The rate of indicated preterm birth varied by gestational age and was highest (39.7%) at 29-32 weeks' gestation. Instrumental preterm births declined over time from 9.5 to 7.8% with a shift from forceps to vacuum use and episiotomy rates declined from 19.7 to 14.8%. CONCLUSIONS: Increases in the reported overall preterm rate (singletons and multiples) were not due to increased delivery of singleton infants. Changes in the management of singleton preterm births were similar to changes observed in term births such as decreasing forceps and episiotomy usage. It may be to time to reassess whether Australian clinicians would be willing to randomise patients to clinical trials of the best method of delivery for preterm infants. PMID- 12755345 TI - Twin pregnancy outcomes for women with gestational diabetes mellitus compared with glucose tolerant women. AB - OBJECTIVE: To examine pregnancy outcomes for women with gestational diabetes mellitus (GDM) and a twin pregnancy compared with glucose tolerant women with a twin pregnancy. DESIGN: Comparison of selected pregnancy outcomes. SETTING: Wollongong, New South Wales, Australia. POPULATION: Women with GDM seen over a 10 year period by an endocrinologist, and women from a selected year of an obstetric database including Wollongong and Shellharbour Hospitals. METHODS: Examination of pregnancy outcome data from the two sources. MAIN OUTCOME MEASURES: Fetal birthweights and method of delivery. RESULTS: There were 28 GDM women with a twin pregnancy from 1229 consecutive referrals (2.3%) of women with GDM for medical management. For comparison there were 29 glucose tolerant women with twin pregnancies evaluable who had delivered over a 1-year period. For the women with GDM and a twin pregnancy there were no significant differences in demographics or outcomes except for a higher rate of elective Caesarean section. CONCLUSION: The higher rate of Caesarean section appeared to be related to the combination of a twin pregnancy and GDM rather than the twin pregnancy or the GDM independently. PMID- 12755346 TI - Changes in body image satisfaction during pregnancy: a comparison of high exercising and low exercising women. AB - OBJECTIVE: This study aimed to compare ratings of body image satisfaction (BIS) from 6 months prepregnancy to 23-30 weeks' gestation for high exercising and low exercising pregnant women. The authors also aimed to assess and compare expectations of BIS for the post-partum period in high and low exercising women. DESIGN: A partial prospective approach was implemented. SAMPLE: A total of 71 healthy pregnant women (40 high exercisers and 31 low exercisers) participated. METHODS: Participants completed a series of questionnaires at 15-22 weeks' gestation and 23-30 weeks' gestation. MAIN OUTCOME MEASURES: There were two main outcome measures. At 15-22 weeks' gestation there was an exercise inventory and two versions of the Body Cathexis Scale (BCS) (retrospective prepregnancy BIS and current BIS). At 23-30 weeks' gestation there was an exercise inventory and two versions of the BCS (current BIS and projected post-partum BIS). RESULTS: At 15 22 weeks' gestation, high exercisers demonstrated significantly higher levels of BIS compared to low exercisers. There were no other significant differences between groups. Within groups, high exercisers were significantly more satisfied with their bodies at 15-22 weeks' gestation compared to 6 months prepregnancy, and expected to be less satisfied with their bodies at 6 weeks' post-partum than they were during pregnancy. Low exercisers demonstrated no significant changes over time. CONCLUSIONS: The findings suggest that women are able to assimilate the bodily changes of pregnancy without a negative shift in BIS. However, women who exercise during pregnancy may respond more favourably to changes in their bodies at early pregnancy compared to women who remain sedentary. PMID- 12755347 TI - Incidence of fetal macrosomia and birth complications in Chinese immigrant women. AB - OBJECTIVE: To compare rates of fetal macrosomia (birthweight > 4000 g) and birth complications in both Chinese women immigrants and Caucasian women for two time periods: 1992 and 1999-2000. POPULATION: Chinese women immigrants and Caucasian women attending the Royal North Shore Hospital and Hornsby Ku-Ring-Gai Hospital in Sydney's northern health region. METHODS: Data used were extracted from the Northern Suburbs Area Health Service OBSTET database. Significance of trends were assessed using chi2 test. RESULTS: The results show a rise in macrosomic babies born to Chinese immigrants from 4% of total Chinese births in 1992 to 9.8% in 1999-2000 (P = 0.02). There was no significant difference in the rate of macrosomia among Caucasian women with respective rates of 11 and 14% for the same periods. The incidence of post-partum haemorrhage increased significantly in both Chinese immigrants and Caucasian women (P < 0.001). CONCLUSION: Australia has a multicultural population and yet the normal ranges defined for many obstetric investigations do not adjust for ethnicity. The application of values derived from a Caucasian population to other ethnic populations may be inappropriate and conceal important pathologies. PMID- 12755348 TI - Outpatient management of abnormal smears. AB - OBJECTIVES: To review the results of the first 403 women treated at the Abnormal Smear and Colposcopy Unit with special reference to the utility, efficacy, acceptability and economy of in-office treatment of cervical lesions by large loop or Fischer cone excision. DESIGN: Retrospective chart review of consecutive patients treated following, referral with an abnormal smear or abnormal cervical morphology, between 1 September 1996 and 1 August 2001. SETTING: Inner city private practice. SAMPLE: A total of 403 consecutive General Practitioner referred women. METHODS: Details of referral smear result, colposcopically directed biopsy result, subsequent treatment type and histological result including assessability, number of specimens submitted, complications and follow up assessment were extracted at chart review. Costs of public hospital inpatient and outpatient care, supplied by the Casemix and Clinical Benchmarking Service, Mater Miseraecordae Public Hospitals (with permission to publish), were compared with Medicare rebates. MAIN OUTCOME MEASURES: A total of 187 women were treated by large loop excision of the transformation zone, and 216 by Fischer cone excision. The number of women who were treated as outpatients under local anaesthetic were 395, while eight patients were treated under general anaesthesia as inpatients. There was poor correlation between referring smear, biopsy and subsequent treatment results. Eight patients had abnormal cytology at follow-up, of whom two have been retreated. Three patients had primary or secondary bleeding requiring treatment and two developed cervical stenosis. Outpatient private practice treatment of women with abnormal smears allows significant savings to the public purse over public or private hospital care. CONCLUSIONS: Outpatient treatment of women with abnormal smears, using the Fischer cone technique, is safe, well accepted, effective and the most cost efficient solution to this public health problem. PMID- 12755349 TI - Role of perineal sonography in the evaluation of patients with stress urinary incontinence. AB - OBJECTIVE: The aim of the study was to determine the role of perineal sonography in diagnosis of stress urinary incontinence. MATERIALS AND METHODS: Thirty patients with stress urinary incontinence and 17 age-matched control patients were included in the study. Perineal sonography was carried out in both groups to evaluate the role of this technique in the diagnosis of stress urinary incontinence. By using the posterior edge of the symphysis pubis as a reference point, posterior urethra-vesical angle (beta angle) and the angle between the vertical axis and urethral axis(alpha angle) were measured at rest and on straining. Bladder neck mobility was evaluated only at the cephalocaudal plane by measuring the desensus diameter. RESULTS: Posterior urethro-vesical angle (beta angle) was found to be significantly different between the study and control groups both at rest and on straining (P < 0.05). The angle between the vertical axis and urethral axis (alpha angle) was found to be significantly different between study and control groups only on straining (P < 0.05). Cephalocaudal distance (desensus diameter) was longer in patients with stress urinary incontinence (P < 0.05). CONCLUSION: Perineal sonography has an important role in diagnosing patients with stress urinary incontinence. PMID- 12755350 TI - A new, simple, safe, effective and cost-effective procedure for genuine stress incontinence: midurethral polypropylene sling. AB - We developed a cost-effective procedure for genuine stress incontinence (GSI) that has the advantages of the tension-free vaginal tape (TVT). The midurethral polypropylene sling procedure (MPS) is carried out under local anaesthesia. A self-fashioned sling (7.5 x 1 cm) was created from a polypropylene mesh with two lengthening polypropylene sutures at the ends. The sutures are carried through the rectus fascia using a needle and the sling is placed around the urethra. Ten patients underwent the MPS and were followed up for a mean of 6.2 months. All patients were cured. The short-term results of the MPS were comparable to those of the TVT. The procedure costs approximately US dollar 9. We conclude that the MPS can be considered as an alternative to the TVT procedure. PMID- 12755351 TI - Malignant mixed mullerian tumour of the ovary: prognostic factor and response of adjuvant platinum-based chemotherapy. AB - OBJECTIVES: The purpose of the present study was to analyse retrospectively the data of a series of patients presenting to our unit with malignant mixed mullerian tumour (MMMT) of the ovary to identify the prognostic factors and relate them to survival. The role of platinum-based chemotherapy in the adjuvant treatment of this tumour was also evaluated. METHODS: All patients diagnosed with MMMT of the ovary from 1987 to 2000 were identified from the gynaecological tumour registry of King George V Hospital, Australia. The effect of clinical and histopathological variables on survival was analysed. The response of platinum based adjuvant chemotherapy after surgery was also evaluated. RESULTS: Twenty patients with MMMT of the ovary were identified. Of the six patients with measurable disease, two (33%) had complete response after adjuvant platinum-based chemotherapy. The median survival of all patients was 8 months, while that of the patients receiving adjuvant platinum-based chemotherapy was 23 months. Women who were older (> 65 years) had a significantly worse survival rate than those who were younger (P = 0.02). The patients with optimal debulking had a better median survival than those with suboptimal debulking, but this difference was not statistically significant (P = 0.21). Sarcomatous component (homologous vs heterologous) was not found to be a significant prognostic factor for predicting survival. CONCLUSIONS: Malignant mixed mullerian tumour of the ovary is a rare and aggressive gynaecological tumour. The current study indicates that patient age was a significant prognostic factor for survival and surgical cytoreduction combined with platinum-based chemotherapy is the most effective management regimen identified to date to treat MMMT of the ovary. PMID- 12755352 TI - Effect of laparoscopic hysterectomy on bladder neck and urinary symptoms. AB - OBJECTIVE: To assess the effect of laparoscopic hysterectomy (LH) on the mobility and position of bladder neck (BN) and urinary symptoms. DESIGN: We assessed the BN and urinary symptoms of 151 patients by introital ultrasonography and questionnaires before and after LH. SAMPLE: One hundred and fifty-one women who underwent LH from June 1999 to June 2001. RESULTS: A significant decrease was noted in the number of women exhibiting one or more urinary symptoms from 81 (53.6%) preoperatively to 58 (38.4%) postoperatively (P < 0.01). The incidence of urinary frequency, mild stress incontinence and nocturia decreased significantly after laparoscopic hysterectomy (P < 0.01). Changes in other urinary symptoms following hysterectomy showed no statistical significance (P > 0.05). During straining, the postoperative position of the BN localised more dorsally (P < 0.01) and the ventral mobility of the BN decreased significantly following surgery (P < 0.05). There was no significant difference in the location of the BN with respect to the pubis at rest and during straining, in the cephalocaudal direction, before and after hysterectomy (P > 0.05). CONCLUSION: Some patients experienced a substantial improvement of preoperative urinary symptoms following LH, partly as a result of a decrease in the hypermobility of BN. PMID- 12755353 TI - Antenatal pelvic organ mobility is associated with delivery mode. AB - OBJECTIVE: Relaxation of pelvic ligaments may facilitate parturition in certain animal species. Biomechanical properties of pelvic connective tissue may also influence progress of labour in the human female. This study was designed to test whether peripheral joint mobility or pelvic organ mobility as measures of connective tissue biomechanical properties are associated with progress in labour and delivery mode. DESIGN: Prospective clinical observational study. SETTING: Tertiary obstetric service. SAMPLE: 200 nulliparous women recruited in antenatal clinic. METHODS: Translabial ultrasound was used to obtain data on third trimester pelvic organ mobility. Upper limb joint mobility was assessed clinically. MAIN OUTCOME MEASURES: Gestational length, length of first and second stage of labour, delivery mode. RESULTS: Pelvic organ mobility was significantly associated with total length of second stage (P = 0.034 to P = 0.002). This was mainly due to the length of passive, not active second stage. There also was a statistically significant association between delivery mode and pelvic organ descent (P = 0.007 to P = 0.001), with the lowest mobility seen in women who required a Caesarean section in second stage. Joint mobility did not correlate with delivery data. CONCLUSION: Third trimester pelvic organ mobility is associated with duration of second stage and delivery mode. PMID- 12755354 TI - Comparability of the amniotic fluid index and single deepest pocket measurements in clinical practice. AB - Two ultrasound techniques, the amniotic fluid index (AFI) and the single deepest pocket (SDP), are currently used to detect oligohydramnios, predict variable decelerations, risk of Caesarean delivery for fetal distress, Apgar scores, umbilical cord artery pH, perinatal mortality, and cerebral palsy. Both techniques poorly identify oligohydramnios. Both techniques identify pregnancies at risk for variable decelerations, low Apgar scores, and Caesarean delivery for fetal distress. Only the SDP is predictive of a compromised fetus-umbilical artery pH, as a stand-alone test, has been correlated with perinatal mortality, and as part of the biophysical profile has been linked to cerebral palsy. This brief communication reviews the comparability of these two techniques and which method, if either, is superior in the identification of oligohydramnios, the predictability of these techniques to identify an adverse pregnancy outcome, and the ability to predict cerebral palsy and perinatal mortality. PMID- 12755355 TI - Management of labour among women with epidural analgesia. AB - To assess current practices in the labour management of low risk primiparous women with epidural analgesia we surveyed delivery suites in New South Wales (NSW) that annually provide at least 100 epidurals to 'standard primipara'. Epidural rates among 'standard primipara' at these hospitals ranged from 14 to 85% (median 46%). Continuous epidural infusion was the most commonly used technique (63%). For 'standard primipara' with an epidural 62% of units usually augmented labour with oxytocin, 89% discontinued the epidural in second stage and 67% had policies of delayed pushing. There is wide variation in epidural availability and in labour management, perhaps reflecting the limited evidence for effective interventions to reduce any unintended effects of epidural analgesia. PMID- 12755356 TI - Fallopian tube carcinoma presenting as cervical growth: the dilemma of skip metastasis. PMID- 12755357 TI - Capillary haemangioma of the uterus: a rare cause of menorrhagia. PMID- 12755358 TI - Hepatic endometriosis: a case report and review of the literature. PMID- 12755359 TI - Birth is not an illness. PMID- 12755360 TI - Early laparoscopy in Australia. PMID- 12755361 TI - Re: Early laparoscopy instruments. PMID- 12755362 TI - Mucosal immunity and microbial pathogenicity. PMID- 12755364 TI - Intestinal M cells and their role in bacterial infection. AB - M cells are located in the epithelia overlying mucosa-associated lymphoid tissues such as Peyer's patches where they function as the antigen sampling cells of the mucosal immune system. Paradoxically, some pathogens exploit M cells as a route of invasion. Here we review our current knowledge of intestinal M cells with particular emphasis on the mechanisms underlying bacterial infection of these atypical epithelial cells. PMID- 12755363 TI - Role of secretory antibodies in the defence against infections. AB - Adaptive immunity mediated by secretory antibodies is important in the defence against mucosal infections. Specific secretory immunoglobulin A (SIgA) can inhibit initial pathogen colonization by performing immune exclusion both on the mucosal surface and within virus-infected secretory epithelial cells without causing tissue damage. Resistance against toxin-producing bacteria such as Vibrio cholerae appears to be particularly dependent on SIgA antibodies. Like natural infections, live topical vaccines or adequate combinations of inactivated vaccines and mucosal adjuvants give rise not only to SIgA antibodies, but also to long-standing serum IgG and IgA responses. The intranasal route of vaccine application could be particularly attractive to achieve this result, but only if successful stimulation is obtained without the use of toxic adjuvants. The degree of protection after vaccination may range from complete inhibition of reinfection to reduction of symptoms. In this scenario it is generally difficult to determine unequivocally the relative importance of SIgA versus serum antibodies. However, infection models in knockout mice strongly support the notion that SIgA exerts a decisive role in protection and cross-protection against a variety of infectious agents. PMID- 12755365 TI - Interaction of Yersinia enterocolitica with epithelial cells: invasin beyond invasion. AB - The chromosomally encoded inv gene product is an outer membrane protein that is functionally expressed in the enteropathogenic Yersinia species Yersinia enterocolitica and Yersinia pseudotuberculosis. Invasin protein is a high affinity ligand for beta1 integrins and especially important in the early phase of intestinal infection for efficient translocation through the M cells located in the follicle-associated epithelium overlying the Peyer's patches. In addition to bacterial internalization, Yersinia invasin mediates proinflammatory epithelial cell reactions. Epithelial cells exhibit immunological functions including production of cytokines thereby signaling to the immune system the presence of invasive or pathogenic bacteria. Several other enteropathogenic bacteria also induce cytokine production in epithelial cells. However, the signaling pathways by which this reaction is accomplished differ for various pathogens. Binding of invasin-expressing Yersinia to beta1 integrin receptors of epithelial cells induces activation of a signal cascade involving Rac1, MAP kinases, activation of the transcription factor NF-kappaB, and the subsequent production of chemotactic cytokines. The Yersinia invasin-triggered inflammatory epithelial cell reaction may lead to the recruitment of phagocytes followed by tissue disruption which may be part of the strategy of the pathogen to promote its dissemination in the host tissue. PMID- 12755366 TI - Shigella interaction with intestinal epithelial cells determines the innate immune response in shigellosis. AB - Shigellae are Gram-negative bacilli that cause bacillary dysentery in humans. This review summarizes current knowledge of Shigella pathogenesis and pathogenicity factors, invasion of epithelial cells, intracellular motility and cell-to-cell spreading, as well as components of the host cell involved in innate immune responses. PMID- 12755367 TI - Mucosal immune response to Tropheryma whipplei. AB - Whipple's disease is a rare infectious disease caused by the ubiquitously occurring Tropheryma whipplei in predisposed persons. Genetic or acquired defects in the mucosal and peripheral immune system become apparent as diminished Th1 immune functions with decreased production of IL-12 and IFN-gamma accompanied by an increased secretion of IL-4. These defects may enable T. whipplei to survive and replicate. The recently established cultivation of the bacterium in HEL cells and the isolation from infected intestinal biopsies enable a multitude of experimental possibilities which may lead to an improved diagnosis as well as understanding of the etiology and pathogenesis of Whipple's disease. PMID- 12755368 TI - Intestinal flora and mucosal immune responses. AB - The normal intestinal flora and the mucosal immune system exist in close spatial proximity. A normal structure and function of both very complex systems is required for health and develops in a constant and interactive process. An abnormal host response to the normal intestinal flora leads to chronic intestinal inflammation. Probiotic bacteria may modulate the intestinal flora and the mucosal immune response and are an effective therapy for remission maintenance of ulcerative colitis and pouchitis. PMID- 12755369 TI - Host defences to Citrobacter rodentium. AB - Citrobacter rodentium is a natural non-invasive bacterial pathogen which infects the distal colon of mice. It uses the same molecular mechanisms of type III secretion as human enteropathogenic and enterohemorrhagic Escherichia coli to colonise the epithelial cells of the gut and is therefore an ideal model to study host-bacterial pathogen interactions in vivo. Infection elicits mucosal inflammation with similarities to inflammatory bowel disease, and so it is a readily accessible model to investigate the relationship between inflammation and anti-bacterial immunity in the gut. PMID- 12755370 TI - Transfer of eukaryotic expression plasmids to mammalian hosts by attenuated Salmonella spp. AB - Transkingdom transfer of DNA from bacteria to other organisms, well established for bacteria, yeast and plants, was recently also extended to mammalian host cells. Attenuated intracellular bacteria or non-pathogenic bacteria equipped with adhesion and invasion properties have been demonstrated to transfer eukaryotic expression plasmids in vitro and in vivo. Here the mucosal application of attenuated Salmonella enterica spp. as DNA carrier for the induction of immune responses towards protein antigens encoded by expression plasmids, their use to complement genetic defects or deliver immunotherapeutic proteins is reviewed. Plasmid transfer has been reported for Salmonella typhimurium, S. typhi and S. choleraesuis so far but clearly other Salmonella strains should be able to transfer expression plasmids as well. Transfer of DNA is effected most likely by bacterial death within the host cell resulting from metabolic attenuation. Since these bacteria remain in the phagocytic vacuole it is unclear how the DNA from such dying bacteria is delivered to the nucleus of infected cells. Nevertheless, the efficiency that has been observed was astonishingly high, reaching close to 100% under certain conditions. Gene transfer in vivo was mainly directed towards vaccination strategies either as vaccination against infectious microorganisms or model tumors. Interestingly, in some cases tolerance against autologous antigens could be broken. In general, this type of immunization was more efficacious than either direct application of antigen, vaccination with naked DNA or using the same bacterium as a heterologous carrier expressing the antigen via a prokaryotic promoter. The ease of generating such vehicles for gene transfer combined with technology validated for mass vaccination programs and the efficacy of induction of protective immune responses makes Salmonella as carrier for mucosal DNA vaccination a highly attractive area for further research and development. PMID- 12755371 TI - Bacterial type III translocation: a unique mechanism for cytosolic display of heterologous antigens by attenuated Salmonella. AB - Upon infection, Gram-negative animal and plant pathogens evade the host immune response by utilizing a specialized protein secretion machinery, known as type III secretion system, for the export of bacterial virulence factors delivered directly into the cytosol of target cells. This unique translocation mechanism can be used for the delivery of large protein fragments derived from immunodominant viral and bacterial heterologous antigens into the MHC class I restricted antigen-processing pathway by attenuated Salmonella carrier vaccines. In orally immunized mice, this novel vaccination strategy results in the induction of pronounced peptide-specific cytotoxic CD8 T cell responses. PMID- 12755372 TI - The role of matrix metalloproteinases (MMPs) in the pathophysiology of chronic obstructive pulmonary disease (COPD): a therapeutic role for inhibitors of MMPs? AB - Chronic obstructive pulmonary disease (COPD) is the collective term describing two separate chronic lung disease diseases: emphysema and chronic bronchitis (1). Initial clinical symptoms are shortness of breath and occasional cough. As the disease progresses difficulties in breathing becomes more pronounced, the cough more persistent and becomes associated with production of a clear sputum. In severe cases there are additional heart complications. The major risk factor for COPD is cigarette smoking. Between 1980 and 1990 there was a 22% increase in the occurrence of the disease with attributed 84,000 deaths in 1990 in the USA (www.nhlbi.nih.gov/health). Current therapies address the symptoms and range from bronchodilators, corticosteroids to oxygen. While there are no effective cures, although the disease can be prevented and progress slowed in many cases by removing the principal risk factor: cigarette smoking. Progression of the disease is associated with degradation of elastin in the walls of the alveoli, resulting in the functional destruction of the these organs. The net increase in proteolytic activity leading to this loss of alveoli function is a growing focus of pharmaceutical efforts for identification of a therapy for the amelioration of this disease. Of specific interest for this review has been the potential roles of members of the MMP family in both the destruction of elastin and the aberrant remodeling of damaged alveoli. An example of such a MMP is Metalloelastase. Metalloelastase (MMP-12) is (as the name suggests) capable of degrading elastin, as well as other extra-cellular matrix components. It is produced predominantly by infiltrating macrophages and appears essential for macrophage migration through extra-cellular matrix (2). Mouse metalloelastase knock-out studies implicate this enzyme as a key mediator in the pathology associated with cigarette smoke induced emhysema (3). There is also associative evidence from human genetic and animal studies suggesting a pathological link with other MMPs, such as MMPs 1,2,3,8 & 9. The evidence for the role of these MMPs in the pathological processes associated with COPD and prospects for MMP inhibitors as the basis for future therapies will be addressed in this review. PMID- 12755373 TI - Luteolin alleviates bronchoconstriction and airway hyperreactivity in ovalbumin sensitized mice. AB - OBJECTIVE AND DESIGN: Asthma is an inflammatory disease of the airways and the current focus in managing asthma is the control of inflammation. In this study, we attempted to investigate the anti-asthmatic potential of a plant derived natural compound, luteolin. MATERIAL: We used a murine model of airway hyperreactivity, which mimicked some of the characteristic features of asthma. Male BALB/c mice (8-9 weeks) were used for this study. TREATMENT: Mice (n = 6) were sensitized by intraperitoneal (i. p.) injection of 10 mg of ovalbumin (OVA) on days 0, 7 and 14 followed by aerosol inhalation (5% OVA) treatments daily beginning from day 19 to day 23. To study its preventive effect, luteolin (0.1, 1.0, and 10 mg/kg body weight; daily) was administered orally during the entire period (0 to 23 day) of sensitization. To study its curative effect, mice were first sensitized and then luteolin (1.0 mg/kg body weight daily) was given orally from day 26 to 32. The airway hyperreactivity, immunoglobulin E (IgE) in the sera, and cytokines (IFN-gamma, IL-4 and IL-5) in the bronchoalveolar lavage fluid (BALF) were measured. RESULTS: Both during sensitization and after sensitization, luteolin, at a dose of 0.1 mg/kg body weight, significantly modulated OVA-induced airway bronchoconstriction and bronchial hyperreactivity (p < 0.05). Luteolin also reduced OVA-specific IgE levels in the sera, increased interferon gamma (IFN-gamma) levels and decreased the interleukin-4 (IL-4) and interleukin-5 (IL-5) levels in the BALF. CONCLUSION: Our study showed that luteolin treatment during and after sensitization significantly attenuated the asthmatic features in experimental mice. Therefore, luteolin could be used either as a lead molecule to identify an effective antiasthma therapy or as a means to identify novel anti-asthma targets. PMID- 12755374 TI - Interleukin-4 production from human basophils is critically dependent on the storage conditions employed prior to stimulation. AB - OBJECTIVE AND DESIGN: Marked variations in the interleukin-4 (IL-4)-producing capacity of basophils can be observed when aliquots from the same cell preparation are kept under different conditions before stimulation. The aim of this study was to identify factors affecting the functional activity of basophils and to determine optimal storage conditions. SUBJECTS: Healthy blood donors. METHODS: Aliquots of purified basophils were kept for different time intervals on ice or at 37 degrees C, in buffer or culture medium, respectively. Following subsequent stimulation with anti-IgE, IL-4 release was determined. RESULTS: Upon storage times up to 4 h, basophils produced more IL-4 when kept at 37 degrees C as compared to 4 degrees C. Surprisingly, buffer was superior compared to culture medium for storage. When the storage time was 20 h, IL-4 release was reduced significantly under all conditions studied. CONCLUSION: The storage conditions considerably affect basophil IL-4 release and thus should taken into account when comparing results. PMID- 12755375 TI - Reactive oxygen species downregulate the expression of pro-inflammatory genes by human chondrocytes. AB - OBJECTIVES: To determine the regulatory effects of reactive oxygen species (ROS) on the expression by human osteoarthritic chondrocytes of interleukin (IL)-1beta, -6 and -8, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) gene in response to interleukin (IL)-1beta or lipopolysaccharide (LPS). METHODS: Human chondrocytes in monolayer culture were incubated for 3 h with ROS generating molecules such as S-nitroso-N-acetyl-D,L-penicillamine (SNAP, 100 microM), 3-morpholinosydnonimine (SIN-1, 100 microM), with chemically synthesised peroxynitrite (ONOO-, 10 microM) or hydrogen peroxide (H2O2, 100 microM). After treatment by ROS, chondrocytes were washed and then cultured for the next 24 h with or without lipopolysaccharide LPS (10 microg/ml) or IL-1beta (1.10(-11) M). IL-1beta, IL-6, IL-8, iNOS and COX-2 gene expression was analysed by real time and quantitative RT PCR. IL-6, IL-8 and prostaglandin (PG) E2 productions were assayed by specific immunoassays. Nitrite was measured in the culture supernatants by the Griess procedure. RESULTS: LPS and IL-1beta stimulated IL 1beta, IL-6, IL-8, iNOS and COX-2 gene expression. SNAP significantly downregulated LPS induced overall gene expressions, whereas SIN-1 had no effect. ONOO- inhibited iNOS and COX-2 gene expression but not that of the cytokine genes. When chondrocytes were incubated with IL-1beta, SIN-1 and ONOO dramatically decreased all gene expressions while SNAP was inefficient. H2O2 treatment inhibited both LPS and IL-1beta induced gene expressions. CONCLUSIONS: These data provide an evidence that ROS may have anti-inflammatory properties by depressing inflammatory gene expression. Further, we demonstrate that ROS effects are dependent on the nature of radical species and the signalling pathway that is activated. These findings should be taken into consideration for the management of antioxidant therapy in treatment of inflammatory joint diseases. PMID- 12755376 TI - Reduced production of hyperalgesic substances by mononuclear cells from aged rats incubated with carrageenan: role of interleukin 2 and prostaglandins. AB - OBJECTIVE AND DESIGN: To compare the production of hyperalgesic substances by cells from aged (A; 24-month) and juvenile (J; 2-month) rats. MATERIAL AND METHODS: 4 x 10(5) purified mononuclear cells from J and A were 2 h-stimulated (test) or not (control) by 250 microg lambda-carrageenan/well. Supernatants (0.1 ml) were intraplantarly (ipl) injected in rat paws and development of mechanical hyperalgesia, in grams, evaluated. Rat interleukin 2 (IL 2) and prostaglandin E2 (PGE2) were also assessed for hyperalgesia development. RESULTS: Test supernatants from A compared with J induced significantly less hyperalgesia (-56 +/- 8.1 and -88.4 +/- 4.6 g, respectively, p < 0.05, ANOVA t test). Local injection of a specific, but not a control, antiserum against IL 2 significantly blocked both pure IL 2- and stimulated supernatants-derived hyperalgesia. In contrast to PGE-like materials, IL 2 content in supernatants was compatible with hyperalgesia development. CONCLUSIONS: Hyperalgesia induced by test supernatants was significantly less intense when derived from aged animals. IL 2 may have accounted for such hyperalgesia. PMID- 12755377 TI - Modulation of airway hyperresponsiveness by thiols in a murine in vivo model of allergic asthma. AB - OBJECTIVE AND DESIGN: Since oxidative stress contributes to the pathogenesis of asthma, this study addressed the question whether supplementing the endogenous antioxidant, glutathione (GSH), would alleviate features of allergic asthma in the mouse. MATERIAL AND METHODS: Ovalbumin-sensitized mice received aerosols of the GSH-donors, glutathione-ethyl ester (GSEt) or N-acetylcysteine, before or during respiratory allergen challenges, or during methacholine challenges given one day after the last allergen challenge. Lung GSH levels were measured shortly after allergen or methacholine challenge. In addition, the effect of GSH supplements on airway hyperresponsiveness and inflammatory cell numbers in the airway lumen was assessed. RESULTS: GSEt decreased allergen-induced airway hyperresponsiveness when given in combination with methacholine. However, when given before or during allergen challenge, both GSH-donors failed to decrease the methacholine-induced airway contractility, change cell numbers in the airway lumen, or increase lung GSH levels. In addition, allergen challenges of sensitized mice did not decrease lung GSH levels. CONCLUSION: In contrast to guinea pigs and humans, allergen challenges in mice does not lead to acute oxidative stress. PMID- 12755378 TI - Analysis of the inflammatory response in the rat paw caused by the venom of Apis melifera bee. AB - OBJECTIVE: This study examines the pro-inflammatory action caused by subcutaneous (s.c.) injection of the bee venom (BV) Apis melifera in the rat paw. METHODS: Male Wistar rats were used. The venom of Apis melifera was injected s.c. into the rat paw and the oedema formation and the activity of myeleperoxidase (MPO) were measured. RESULTS: Subcutaneous injection of BV caused dose-and time-dependent paw oedema (ED50 of 1.5 microg/paw) with peak at 30 min. The MPO activity increased about 1.6, 4.2 and 8.9 folds at 0.5, 4 and 6 h after s.c. injection of BV. The mast cell degranulating drug 48/80, pyrilamine or metysergide, inhibited BV-mediated oedema formation (88, 62 and 96%, respectively). Likewise, L-NAME, the NK1 antagonist FK 888, the B1 des-Arg9-[Leu8]-BK or B2 kinin antagonist Hoe 140 also antagonised the paw oedema induced by BV (60, 59, 49, and 49%, respectively). SR48968 and SR14280, respectively NK2 and NK3 antagonists and also indomethacin, inhibited by 31, 29 and 22%, respectively BV-induced oedema formation. In contrast, the PAF antagonist WEB 2086 or valeryl salycilate, did not affect the BV-induced paw oedema. The levels of MPO were inhibited by compound 48/80, cyproheptadine, Hoe 140, or by des-Arg9[Leu8]-BK (85, 61, 59, and 53%, respectively) measured 6 h after. CONCLUSION: These results indicate that the BV from Apis melifera causes a marked dose-and time-dependent oedema formation in the rat paw, an effect that is accompanied by intense leukocyte migration. The pro-inflammatory response induced by BV is mediated by several mechanisms, namely the release of histamine and/or serotonin from mast cells, activation of H1 histamine receptor, production of nitric oxide, the involvement of kinins through the activation of B1 and B2 receptors, and also tachykinins acting at NK1 receptor or and to a lesser extent at NK2 and NK3 receptors. PMID- 12755379 TI - Enhanced histamine metabolism: a comparative analysis of collagenous colitis and food allergy with respect to the role of diet and NSAID use. AB - OBJECTIVES AND DESIGN: To compare clinical data and histamine metabolism of patients with collagenous colitis with those of food allergy. METHODS: In 17 patients with collagenous colitis, clinical findings (diarrhoea, abdominal pain) were recorded. Plasma (for histamine) and 12-h-urine (for histamine and n methylhistamine, all measured by RIA) were collected during two days with an unrestricted diet followed by two days with an hypoallergenic. The clinical data and measured values were compared with those from patients with confirmed food allergy (n = 21) and controls (n = 41). RESULTS: Patients with collagenous colitis were found to present with significantly more liquid stools than patients with food allergy (p < 0.001) during both types of diet, but they did not experience more abdominal pain. N-methylhistamine in 12-h-urine was significantly increased during both types of diet in patients with collagenous colitis and food allergy when compared with controls (p < 0.001 for all). Patients with food allergy--but not those with collagenous colitis--showed a significant decrease of severity of pain (p < 0.05) when the diet was changed to the elimination protocol. CONCLUSION: Histamine is extensively produced and metabolised in patients with collagenous colitis. In contrast to food allergy, the allergenic potency of the administered food seems not to influence histamine production in collagenous colitis. However, histamine metabolism corresponds with the clinical activity in both patients with food allergy and collagenous colitis. PMID- 12755380 TI - Impact of topoisomerase II inhibition on cytokine and chemokine production. AB - OBJECTIVE AND DESIGN: In systemic inflammatory diseases such as rheumatoid arthritis, cytokines and chemokines are deeply involved in the development of the disease manifestations. Etoposide is a cytostatic drug, known to deplete the monocyte population in mice and rabbits. We have recently shown that suboptimal doses have a disease-ameliorating effect in collagen II induced arthritis in the absence of monocyte depletion. Anti-arthritic properties parallelled with almost total eradication of production of specific collagen II antibodies. The aim of the present study was to investigate ex vivo and in vitro the function of the mononuclear cells and their production of B cell stimulating cytokines following exposure to etoposide, a topoisomerase II inhibitor. MATERIALS AND METHODS: Spleen cells from mice treated during four weeks with etoposide were cultured and the supernatants were analyzed with respect to content of TNF and IL-6. In addition, cells from the murine macrophage cell clone IC-21 were exposed to etoposide and the production of IL-6, using a bioassay, and the production of TNF, MIP-1alpha, RANTES, and IL-1beta, using sandwich ELISAs, was determined. RESULTS: Spleen cells from etoposide-treated mice secreted lower amounts of IL-6 and TNF as compared to the control animals. In addition, in vitro etoposide exposed macrophages showed reduced capacity to produce TNF, IL-6 and MIP-1alpha. CONCLUSION: Our results indicate that inhibition of topoisomerase II downregulated the function of monocytes. Owing to its immunoregulatory properties, use of etoposide is suggested in treatment of chronic inflammatory diseases. PMID- 12755381 TI - Pyrethroid insecticides influence the signal transduction in T helper lymphocytes from atopic and nonatopic subjects. AB - OBJECTIVE AND DESIGN: Pyrethroids are claimed to have a low human toxicity with some neuro- and immunotoxicity. The objective of this study was to investigate the immunotoxicological properties of six commercially used pyrethroids, including natural pyrethrum and synergist piperonyl-butoxide (PBO). MATERIAL AND METHODS: PHA-stimulated cultures of T-helper lymphocytes and blood basophil incubates from nonatopic and atopic patients (IgE > 1000 IU) provided cytokine and histamine determination. Western blot analysis was used for the measurement of Th2-specific signal transducer and activator of transcription-6 (STAT6). Pyrethroids and xenobiotics were added 4 h post-plating. RESULTS: We demonstrated that interferon-gamma (IFN-gamma) production and expression was correlated with lymphocyte proliferation, however, interleukin-4 (IL-4) was down-regulated at the end of the 3 day culture. Atopics showed significantly higher IL-4 activity than nonatopics. Pyrethroids inhibited IFN-gamma and IL-4 in both groups at around 10( 5) M. Only fenvalerate and S-bioallethrin combined with 10-fold PBO in the atopic enriched blood basophil incubates caused a weak but significant increase in histamine release. Histamine acted bidirectionally on STAT6, but pyrethroids inhibited the intracellular Th2-specific STAT6 more effectively in atopics than in nonatopics. CONCLUSION: It can be suggested that pyrethroids inhibit signal transduction in human lymphocytes ex vivo, and do not act via lymphocyte influencing histamine release. PMID- 12755382 TI - Amelioration of hyperalgesia by kinin receptor antagonists or kininogen deficiency in chronic constriction nerve injury in rats. AB - OBJECTIVE: The present study was designed to examine the involvement of bradykinin in thermal and mechanical hyperalgesia induced by chronic constriction nerve injury (CCI) using B1 and B2 receptor antagonists and mutant kininogen deficient rats. METHODS: Sprague-Dawley (SD) rats and Brown Norway (B/N-) rats given CCI treatment on day 0, were used as a model of neuropathic pain. Either a kinin B1 antagonist des-Arg9-[Leu8]-bradykinin or the receptor B2 antagonist HOE 140 was constantly infused into the left jugular vein of SD rats on days 15 to 22 after CCI. Vehicle-treated rats and sham-operated rats without nerve injury were also prepared as controls. In all rats, we observed pain behavior, and measured the latency period of paw withdrawal from the thermal stimuli and, with von Frey filaments, the mechanical pain threshold, before surgery and on days 14 and 22 after CCI. B/N-Katholiek rats, which congenitally lack plasma kininogen and release no kinin, were also tested for hyperalgesic parameters. Expression of kinin receptor mRNA in the dorsal root ganglia was detected by RT-PCR. RESULTS: Most of the rats (88%) showed some pain behavior, which was reduced to 67% by a B1 antagonist and to 57% by a B2 antagonist infused between days 15 to 22. Thermal hyperalgesia was significantly reduced from 7.25 +/- 0.41 sec (mean +/- SEM) to 8.36 +/- 0.41 sec in paw withdrawal latency on day 22 by a B1 antagonist and from 7.24 +/- 0.19 sec to 8.23 +/- 0.21 sec by a B2 antagonist (P < 0.05). Mechanical hyperalgesia was also ameliorated from 0.02 +/- 0.007 g force to 0.16 +/- 0.08 g force in pain threshold by a B1 antagonist and from 0.03 +/- 0.007 g force to 0.10 +/- 0.003 g force on day 22 by a B2 antagonist. Moreover, deficient B/N-Katholiek rats showed a low incidence of thermal and mechanical hyperalgesia on day 14. Expression of both B1 and B2 receptor mRNAs was detected in the lumbar dorsal ganglia ipsilateral to the site of the nerve injury. CONCLUSION: These data suggests that kinin were at least partly involved in yielding nociceptor hypersensitivity up to day 14 after CCI. Bradykinin and its B1 and B2 receptors were involved in the maintenance of hyperalgesia. PMID- 12755383 TI - Administration of the non-steroidal anti-inflammatory drug ibuprofen increases macrophage concentrations but reduces necrosis during modified muscle use. AB - OBJECTIVE: To test the hypothesis that ibuprofen administration during modified muscle use reduces muscle necrosis and invasion by select myeloid cell populations. METHODS: Rats were subjected to hindlimb unloading for 10 days, after which they experienced muscle reloading by normal weight-bearing to induce muscle inflammation and necrosis. Some animals received ibuprofen by intraperitoneal injection 8 h prior to the onset of muscle reloading, and then again at 8 and 16 h following the onset of reloading. Other animals received buffer injection at 8 h prior to reloading and then ibuprofen at 8 and 16 h following the onset of reloading. Control animals received buffer only at each time point. Quantitative immunohistochemical analysis was used to assess the presence of necrotic muscle fibers, total inflammatory infiltrate, neutrophils, ED1+ macrophages and ED2+ macrophages at 24 h following the onset of reloading. RESULT: Administration of ibuprofen beginning 8 h prior to reloading caused significant reduction in the concentration of necrotic fibers, but increased the concentration of inflammatory cells in muscle. The increase in inflammatory cells was attributable to a 2.6-fold increase in the concentration of ED2+ macrophages. Animals treated with ibuprofen 8 h following the onset of reloading showed no decrease in muscle necrosis or increase in ED2+ macrophage concentrations. CONCLUSION: Administration of ibuprofen prior to increased muscle loading reduces muscle damage, but increases the concentration of macrophages that express the ED2 antigen. The increase in ED2+ macrophage concentration and decrease in necrosis may be mechanistically related because ED2+ macrophages have been associated with muscle regeneration and repair. PMID- 12755384 TI - Effect of lipopolysaccharide on D-fructose transport across rabbit jejunum. AB - OBJECTIVE: To investigate alterations in the transport of D-fructose across the rabbit jejunum when the gut is exposed in vitro to lipopolysaccharide (LPS), an endotoxin causative agent of sepsis. MATERIALS AND METHODS: D-fructose intestinal transport was assesed employing three techniques: sugar uptake measurements in rings of everted jejunum (micromol/D-fructose/ml cell water), transepithelial flux measurements in Ussing-type chambers (micromol D-fructose/cm2/h) and transport assays in preparation of brush border membrane vesicles (pmoles D fructose/mg protein). Samples were taken from the bathing solution and from the extracts of the tissue for radioactivity counting. RESULTS: Adding LPS (3 microg/ml) to tissue decreased the uptake and mucosal to serosal flux of 5 mM D fructose across the enterocyte. LPS did not modify sugar uptake across brush border membrane vesicles. The inhibitory effect of LPS was suppressed by W-13 (5 x 10(-6) M), a Ca-calmodulin antagonist, and staurosporine (10(-7) and 10(-6) M) and GF-109203X (10(-6) M) a nonselective and selective protein kinase C (PKC) inhibitor respectively. Tumor Necrosis Factor (TNF-alpha), an immunoregulatory cytokine involved in septic responses occurring during bacterial infection at concentrations 3 x 10(-4) to 3 microg/ml, did not affect the sugar transport. CONCLUSIONS: LPS can inhibit the intestinal uptake of D-fructose across the rabbit jejunum in vitro by intracellular processes related to PKC and calmodulin protein. PMID- 12755385 TI - Histamine uptake into non-neuronal brain cells. PMID- 12755386 TI - Nitric oxide modulates the inhibitory effect of cannabinoids on the immunological activation of guinea pig mast cells. PMID- 12755387 TI - Mast cell-derived vascular endothelial growth factor (VEGF) and microvascular density in diabetic placentae. PMID- 12755388 TI - Pyrethroids do not show relevant ex vivo histamine releasing potency in human basophils. PMID- 12755389 TI - Diagnostic use of mucosa oxygenation and histamine release experiments in patients with gastrointestinally mediated allergy (GMA). PMID- 12755390 TI - A23187-activated mast cells affect intestinal function in the pig proximal colon- role for prostaglandins. PMID- 12755391 TI - Differential inhibition of antigen- and anti-immunoglobulin-induced histamine release from human leucocytes by D-penicillamine and dithiothreitol. PMID- 12755392 TI - Histamine, dithiaden and human polymorphonuclear leukocytes in vitro. PMID- 12755393 TI - The effects of histamine on the lipopolysaccharide-induced metallothionein-I mRNA expression in the mouse spleen. PMID- 12755394 TI - Effects of the SOD mimetic, M40403, on prostaglandin production in an in vivo model of ischemia and reperfusion in rat heart. PMID- 12755395 TI - Effect of protease-activated receptors on guinea-pig ileum contraction in vitro. PMID- 12755396 TI - Brain histamine turnover and seizure behaviour in two mouse strains: preliminary findings. PMID- 12755397 TI - Increased density of H1 histamine receptors in brain regions of rats with absence epilepsy. PMID- 12755398 TI - Identification of novel H3 receptor (H3R) antagonists with cognition enhancing properties in rats. PMID- 12755399 TI - Pro-cognitive effect of a selective histamine H1-receptor agonist, 2-(3 trifluoromethylphenyl)histamine, in the rat object recognition test. PMID- 12755400 TI - Suppressed weight gain following chronic I-histidine loading--involvement of neuronal histaminergic system? PMID- 12755401 TI - Alcoholics show altered histaminergic neurotransmission in several cortical areas -preliminary report. PMID- 12755402 TI - N,N-diethyl-2-[4-(phenylmethyl)phenoxy]ethanamine increases the permeability of primary mouse cerebral endothelial cell monolayers. PMID- 12755404 TI - Role of putative phosphorylation sites in down-regulation of histamine H1 receptors. PMID- 12755403 TI - Central histamine-induced reversal of haemorrhagic shock in rats--a comparison with the pressor effect of peripheral adrenergic receptor stimulation. PMID- 12755405 TI - In vitro pharmacological properties of two novel non-imidazole H3 receptor (H3R) antagonists. PMID- 12755406 TI - D-alanine piperazine-amides: novel non-imidazole antagonists of the histamine H3 receptor. PMID- 12755407 TI - H1 antagonists: receptor affinity versus selectivity. PMID- 12755408 TI - Histamine H1-agonist properties of histaprodifen derivatives on guinea-pig isolated trachea and ileum. PMID- 12755409 TI - Mouse embryonic stem cells express histidine decarboxylase and histamine H1 receptors. PMID- 12755410 TI - Histamine antilipolytic action in rat adipocytes: comparison with the effect of tyramine. PMID- 12755411 TI - Standardised evaluation of urinary excretion of N-tele-methylhistamine in different periods of age in a healthy population. PMID- 12755412 TI - Phosphorothioate antisense oligodeoxynucleotides against histidine decarboxylase: a study in mouse mammary epithelial cell cultures. PMID- 12755413 TI - Effects of unfractionated and low molecular weight heparins on diamine oxidase release. PMID- 12755414 TI - Simultaneous purification of the histamine degrading enzymes diamine oxidase and histamine N-methyltransferase from the same tissue. PMID- 12755415 TI - Alterations in plasma amine oxidase activities in a compartment syndrome model. PMID- 12755416 TI - Analysis of genetic polymorphisms of enzymes involved in histamine metabolism. PMID- 12755417 TI - Genetic polymorphisms of histamine degrading enzymes: from small-scale screening to high-throughput routine testing. PMID- 12755418 TI - High expression of semicarbazide-sensitive amine oxidase genes AOC2 and AOC3, but not the diamine oxidase gene AOC1 in human adipocytes. PMID- 12755419 TI - Expression of histidine decarboxylase in human colonic cancer cells and adenomatous polyps. PMID- 12755420 TI - A practical approach to anthelmintic resistance. PMID- 12755421 TI - Excessive airway mucus in horses with pulmonary disease: is it caused by mucus overproduction, decreased clearance or both? PMID- 12755422 TI - Practice is alive with the sounds of horses. PMID- 12755423 TI - The sacroiliac joints: evaluation using nuclear scintigraphy. Part 1: The normal horse. AB - REASONS FOR PERFORMING STUDY: Sacroiliac (SI) disease is recognised as a cause of poor hindlimb action but differential diagnosis is often difficult. HYPOTHESES: That in clinically normal horses there would be a significant difference in the ratio of radiopharmaceutical uptake (RU) between the fifth lumbar vertebra (L5) and each tuber sacrale (TS) and between L5 and each SI joint; and that these ratios would alter with age, but ratios would be bilaterally symmetrical. METHODS: Dorsal scintigraphic images of the SI region of 15 horses, selected randomly from the clinic database, were analysed by 2 of the authors, comparing noncorrected and motion-corrected images. To determine scintigraphic anatomy, the scintigraphic images of 10 Thoroughbred horses were superimposed over a ventrodorsal radiographic image of an isolated pelvis. Dorsal scintigraphic images of 40 clinically normal horses age 3-16 years were evaluated using subjective examination, profile analysis and quantification using regions of interest. RESULTS: The tubera sacrale were seen as 2 well-defined oval regions immediately to the left and right of the midline, abaxial to which were larger, approximately oval areas with less RU, representing uptake in the SI joints. The definition between the SI region and the TS was more obvious in younger horses. Nonmotion-corrected images were often not of diagnostic quality or could be misinterpreted as abnormal. There were significant differences in RU between the TS and SI joints compared to L5, and decreased RU in the tubera sacrale with increasing age, but no change of uptake in the SI joint region and no effect of gender on RU. There was a high degree of left-right symmetry of the TS and SI joint regions. CONCLUSIONS AND POTENTIAL RELEVANCE: The scintigraphic images of horses with suspected sacroiliac joint disease should be compared with images of normal horses of comparable age. In normal horses, there was a high degree of symmetry; therefore, marked left-right asymmetry is likely to be abnormal. PMID- 12755424 TI - The sacroiliac joints: evaluation using nuclear scintigraphy. Part 2: Lame horses. AB - REASONS FOR PERFORMING STUDY: Sacroiliac (SI) joint disease is difficult to diagnose definitively on clinical grounds. HYPOTHESES: Lameness not attributable to SI joint pain may result in asymmetrical uptake of radiopharmaceutical in the tubera sacrale and SI joints; and that horses with clinical signs suggestive of sacroiliac joint disease would have differences in the ratios of radiopharmaceutical uptake between the fifth lumbar vertebra (L5) and either the tubera sacrale or SI joints compared with normal horses. OBJECTIVES: To determine whether differences in radiopharmaceutical activity could identify individuals with presumed SI joint region pain. METHODS: The scintigraphic appearance of the pelvic region of 234 horses (Group I) with lameness unrelated to the SI joints was compared with that for 40 normal horses (Group N) in full work and that for 41 horses with suspected SI joint disease (Group II). The effect of age, sex, lame limb and discipline were assessed in Group I. Motion-corrected scintigraphic images were assessed grossly, using profile analysis and by comparing ratios of uptake of mean count per pixel between L5 and each of the left (L) tuber sacrale (TS), right (R) TS, LSI and RSI joints. Left-right symmetry was compared. RESULTS: Lame horses (Group I) had greater asymmetry of radiopharmaceutical uptake in the tubera sacrale compared with normal horses. Horses with right hindlimb lameness had a larger RSI/LSI ratio compared to normal horses. In Group I L5/LTS, L5/RTS and L5/RSI increased with age, and there was a trend for L5/LSI to increase with age. In horses with presumed sacroiliac joint disease (Group II), L5/LTS, L5/RTS and L5/RSI were all greater compared with normal horses. Detection of marked left-right asymmetry by quantitative analysis or profile analysis was helpful in discriminating between those horses with clinical signs compatible with SI joint disease and either normal horses or those lame due to another cause. CONCLUSIONS AND POTENTIAL RELEVANCE: Scintigraphic evaluation of the SI region is useful to identify SI joint disease in combination with other clinical signs supportive of the diagnosis. However, diagnosis should not be based on this alone because of some degree of overlap in the range of radiopharmaceutical uptake between horses with SI joint disease and both normal horses and those with other causes of lameness. PMID- 12755425 TI - Pain associated with the sacroiliac joint region: a clinical study of 74 horses. AB - REASONS FOR PERFORMING STUDY: There has been no large study of horses with suspected sacroiliac (SI) joint region pain in which the clinical diagnosis has been supported by either abnormal radiopharmaceutical activity in the SI joint region or by periarticular infiltration of local anaesthetic solution. OBJECTIVES: To describe the clinical features of horses with SI joint region pain, to document the age, breed, sex, discipline, size and conformation of affected horses and to compare these with the author's (SD) normal case population and to document the results of infiltration of local anaesthetic solution around the SI joint region. METHODS: Horses were selected for inclusion in the study based upon the exclusion of other causes of lameness or poor performance, together with clinical signs suggestive of SI joint pain and abnormal radiopharmaceutical activity in the SI joint region and/or a positive response to periarticular infiltration of local anaesthetic solution. RESULTS: Sacroiliac joint region disease was identified in 74 horses between November 1997 and March 2002. Dressage and showjumping horses appeared to be at particular risk (P < 0.001). Affected horses were generally slightly older than the normal clinic population (P < 0.0001), taller at the withers (P < 0.0001) and of greater bodyweight (P < 0.01). There was a significant effect of breed (P < 0.001), with a substantially higher proportion of Warmblood horses (51%) in the SI pain group compared to the normal clinic population (29%). There was no correlation between conformation and the presence of SI joint region pain. The tubera sacrale appeared grossly symmetrical in most (95%) horses. Poor development of the epaxial muscles in the thoracolumbar region and asymmetry of the hindquarter musculature were common. Twenty-six horses (35%) showed restricted flexibility of the thoracolumbar region and 10 (16%) had an exaggerated response to pressure applied over the tubera sacrale. Fourteen horses (19%) were reluctant to stand on one hindlimb for prolonged periods. The majority of horses (75%) had a straight hindlimb flight and only 18% moved closely behind or plaited. In all horses restricted hindlimb impulsion was the predominant feature; invariably this was most obvious when the horse was ridden. Stiffness, unwillingness to work on the bit and poor quality canter were common. Sacroiliac joint region pain was seen alone (47%), or in conjunction with thoracolumbar pain (16%), hindlimb lameness (20%), forelimb lameness (7%) or a combination of problems (10%). Seventy-three horses (99%) had abnormalities of the SI joint region identified using nuclear scintigraphy. Infiltration of local anaesthetic solution around the SI joint region produced profound improvement in gait in all 34 horses in which it was performed. CONCLUSIONS AND POTENTIAL RELEVANCE: Careful clinical examination combined with scintigraphic evaluation of the SI joint region and local analgesia can enable a more definitive diagnosis of SI joint region pain than has previously been possible. PMID- 12755426 TI - Management of drug-resistant cyathostominosis on a breeding farm in central North Carolina. AB - REASONS FOR PERFORMING STUDY: Possible anthelmintic resistance on a breeding farm where a rapid rotation anthelmintic programme had been implemented for 9 years was investigated. Cyathostomins resistant to fenbendazole and pyrantel were documented by faecal worm egg count reduction test (FWECRT). OBJECTIVES: To 1) manage small strongyle transmission in a herd of horses in which resistance to both pyrantel pamoate and fenbendazole was identified and thereby reduce the risk of clinical disease in the individual animal, 2) monitor the change in resistance patterns over time and 3) monitor the efficacy of ivermectin over the study period. METHODS: Targeted ivermectin treatment of horses on the farm was instituted for mature horses with faecal worm egg counts (FWEC) > 200 eggs/g (epg) and for horses < age 2 years with FWEC > 100 epg. RESULTS: Over a 30 month period, targeted ivermectin treatment achieved acceptable control in mares, as judged by FWEC, and improved control of patent cyathostome infection in consecutive foal crops. Egg reappearance time (ERT) after treatment with ivermectin was < 8 weeks in mares and foals more frequently in the second year of the study than in the first year. Numbers of anthelmintic treatments were reduced by 77.6 and 533% in the mare and foal group, respectively. CONCLUSIONS: Targeted ivermectin treatment may be an economically viable method of managing multiple drug resistant cyathostominosis. POTENTIAL RELEVANCE: Use of ivermectin should be monitored closely for development of resistance. PMID- 12755427 TI - Mucin genes in horse airways: MUC5AC, but not MUC2, may play a role in recurrent airway obstruction. AB - REASONS FOR PERFORMING STUDY: Increased mucin gene expression may be an important cause of mucus accumulation observed in recurrent airway obstruction (RAO) affected horses. To date, however, no mucin gene sequences are available for the horse. OBJECTIVES: To identify equine homologues of gel-forming mucins and investigate their expression at different airway generations of healthy and RAO affected horses. METHODS: Two equine homologues were identified by cloning and sequencing fragments of equine (eq)MUC5AC and eqMUC2. RESULTS: Semiquantitative RT-PCR on RNA from airways (generations 1, 5, 10, 15; small airways and parenchyma), stomach (glandular), and colon revealed that eqMUC5AC is expressed in equine stomach and in all of the airway samples. In contrast, eqMUC2 steady state mRNA levels were detected in colon and very faintly in stomach, but not in airway tissue. EqMUC5AC expression was also compared to that of ZO-1, a tight junction protein, and eqMUC5AC/ZO-1 ratios were higher in RAO-affected compared to control horses at all airway generations. CONCLUSIONS: That eqMUC5AC is expressed in horse airways, but any expression of MUC2 is undetectable and unlikely to be of physiological consequence. POTENTIAL RELEVANCE: EqMUC5AC up regulation may be a primary mechanism responsible for mucus hypersecretion and accumulation in RAO. PMID- 12755428 TI - Investigations into the role of the thyrohyoid muscles in the pathogenesis of dorsal displacement of the soft palate in horses. AB - REASONS FOR PERFORMING STUDY: Contributes to the understanding of the pathogenesis of dorsal displacement of the soft palate during exercise so that management of this condition could be enhanced. HYPOTHESIS: That the thyrohyoid muscles play an important role in the stability of the laryngo-palatal relationship and that dysfunction of these muscles leads to dorsal displacement of the soft palate (DDSP) during exercise. METHODS: Ten horses were exercised on a high-speed treadmill under 4 different treatment conditions: control conditions (n = 10), after resection of thyrohyoid muscles (TH, n = 10), after sham treatment (n = 5), or after restoration of function of the thyrohyoid muscles with surgical sutures (prosthesis-treatment, n = 6). During trials, the following determinations were made: videoendoscopy of the upper airway, gait frequency and pharyngeal and tracheal static pressures. RESULTS: None of the 10 horses developed DDSP during 2 separate treadmill-exercise trials under the control conditions. Seven of the 10 horses developed DDSP after resection of the TH muscles, 4 of 5 of these horses still experienced DDSP after sham-treatment, but 5 of 6 horses no longer experienced DDSP at exercise after the prosthesis treatment. There were significant anomalies in airway pressures, respiratory frequency, and occurrence of DDSP in both the TH resection and sham-treatment conditions compared to control conditions. In contrast, no statistical differences were noted in any of the parameters measured between the prosthesis treatment and control conditions. CONCLUSIONS: That the function of the TH muscles is important to the stability of the laryngo-palatal relationship and plays a role in the pathophysiology of exercise-induced DDSP. POTENTIAL RELEVANCE: Management of horses with DDSP could be enhanced by restoring the function of the TH muscles. PMID- 12755429 TI - Spectral analysis of respiratory noise in horses with upper airway disorders. AB - REASONS FOR PERFORMING STUDY: It has long been recognised that the production of abnormal respiratory sounds by horses during exercise is frequently associated with upper airway obstructions. Respiratory acoustic measurements have shown promise in investigation of upper airway disorders in man and, more recently, in horses with experimentally-induced obstructions. OBJECTIVES: To evaluate sounds from exercising horses with naturally occurring dynamic obstructions of the upper respiratory tract and to compare these with those from normal horses in order to determine whether different obstructions produce characteristic spectral patterns. METHODS: The audio signal, airflow and videoendoscopic images were recorded simultaneously during an incremental exercise test on a high-speed treadmill. RESULTS: Spectral analysis of the audio signal showed marked differences between control and clinically afflicted horses. Dorsal displacement of the soft palate was characterised by a narrow low frequency (20-80 Hz) peak during expiration. Horses with dynamic laryngeal collapse produced inspiratory sounds characterised by a broad band high frequency spectral component in the range 1.1-2.7 kHz. CONCLUSIONS AND POTENTIAL RELEVANCE: Spectral analysis of respiratory sounds in horses has potential as a diagnostic technique for field use especially when facilities for high-speed treadmill assessment are not practicable. PMID- 12755430 TI - Tenosynovitis of the digital flexor tendon sheath and annular ligament constriction syndrome caused by longitudinal tears in the deep digital flexor tendon: a clinical and surgical report of 17 cases in warmblood horses. AB - REASONS FOR PERFORMING STUDY: Inflammation of the digital flexor tendon sheaths is a chronic and nebulous condition often unresponsive to medical and surgical treatment. OBJECTIVES: To evaluate the incidence of longitudinal tears (LT) as the underlying cause of chronic tenosynovitis and annular ligament constriction syndrome (ALCS) in warmblood horses. METHODS: The records of 25 horses with chronic tenosynovitis and ALCS in which tenoscopical inspection of the digital flexor tendon sheath (DFTS) was performed between 1999-2000 were reviewed. Of 25 horses, 17 were diagnosed with an LT in the deep digital flexor tendon (DDFT). All horses had a history of distension of the DFTS and/or signs of an ALCS. All cases presented typical signs of a chronic inflamed DFTS on ultrasonography and 11 horses showed ultrasonographic changes (echogenic material or an irregular outlining) at the lateral or medial border of the DDFT. The diagnosis of LTs of the DDFT was established in all cases by tenoscopy. Surgical treatment consisted of removal of the torn collagen fibrils using a mechanical resector and decompressing the fetlock canal by a transection of the palmar annular ligament (PAL) using a hook knife under tenoscopic control. RESULTS: Ten horses became sound and resumed their previous level of work, 3 horses remained lame, 4 horses returned to previous level of work but needed intrasynovial treatment of the DFTS and reduced competition frequency to remain sound. CONCLUSIONS: Horses presented with chronic inflamed DFTS and/or ALCS might suffer from LTs in the DDFT; however, the diagnosis cannot be established with absolute certainty using only ultrasonography. Longitudinal tears should be suspected if ultrasonographic changes are present lateral or medial to the border of the DDFT but tenoscopical examination of the tendon sheath is essential to establish an accurate diagnosis and an effective treatment. POTENTIAL RELEVANCE: The presence of these LTs might explain why some cases of chronic tenosynovitis of the digital flexor tendon sheath and/or ALCS do not respond on surgical transection of the PAL alone without tenoscopy. PMID- 12755431 TI - Idiopathic mucosal lesions of the arytenoid cartilages of 21 Thoroughbred yearlings: 1997-2001. AB - REASONS FOR PERFORMING STUDY: Mucosal ulcers and, occasionally, small granulomas on the axial surface of one or both arytenoid cartilages have been found in TB yearlings presented for post sale endoscopic examination. OBJECTIVES: To determine the incidence, endoscopic characteristics and outcome of a group of Thoroughbred yearlings affected with mucosal ulcers and granulomas of the arytenoid cartilage. HYPOTHESIS: The incidence of mucosal ulceration of the arytenoid cartilages of yearling Thoroughbreds is relatively high compared to other upper airway abnormalities; and that the majority of mucosal ulcers heal uneventfully, although a small percentage may progress to a granuloma and, less commonly, to arytenoid chondropathy. METHODS: The findings of post sale, upper airway endoscopic examinations of 3312 Thoroughbred yearlings, during a 5 year period, were reviewed, including those abnormalities listed in the conditions of sale and others not listed but considered likely to cause airway obstruction. Information obtained from the medical record of horses that had mucosal ulceration or granuloma of the arytenoid cartilage included the location and size of the lesion(s), sex of the affected horse and the presence and nature of other concurrent abnormalities of the upper portion of the respiratory tract. Additional information included treatment and results of follow-up, endoscopic examination by the authors or attending veterinarian. RESULTS: Mucosal lesions were seen in 0.63% of yearlings evaluated, which represented the most common, documented condition of the upper portion of the respiratory tract. The mucosal ulcers of 15 of 19 horses were considered to have healed without complication during follow-up examination; one of the 19 horses was lost to follow-up. Two horses affected with bilateral, arytenoid mucosal ulceration developed a granuloma at each site of ulceration. One horse developed a granuloma at a site of ulceration and, subsequently, arytenoid chondropathy. CONCLUSIONS: Arytenoid mucosal ulceration in sales yearlings was a relatively commonly encountered abnormality and a small percentage progressed to granuloma or chondropathy. POTENTIAL RELEVANCE: The mucosa of the arytenoid cartilage, particularly at the rostral margin of the vocal process, should be examined carefully during endoscopic examination of the upper portion of the respiratory tract of Thoroughbred yearlings presented for sale. Because a small percentage of mucosal ulcers may progress to granuloma or, less commonly, chondropathy, identification of mucosal ulcers of the arytenoid cartilage seen during post sale, endoscopic examination warrants notification to the purchaser and sales company. Medical therapy of affected horses should be considered and follow-up endoscopic examination performed to determine if the lesion has healed. PMID- 12755432 TI - Cardiopulmonary, blood and peritoneal fluid alterations associated with abdominal insufflation of carbon dioxide in standing horses. AB - REASONS FOR PERFORMING STUDY: Abdominal insufflation is performed routinely during laparoscopy in horses to improve visualisation and facilitate instrument and visceral manipulations during surgery. It has been shown that high-pressure pneumoperitoneum with carbon dioxide (CO2) has deleterious cardiopulmonary effects in dorsally recumbent, mechanically ventilated, halothane-anaesthetised horses. There is no information on the effects of CO2 pneumoperitoneum on cardiopulmonary function and haematology, plasma chemistry and peritoneal fluid (PF) variables in standing sedated horses during laparoscopic surgery. OBJECTIVES: To determine the effects of high pressure CO2 pneumoperitoneum in standing sedated horses on cardiopulmonary function, blood gas, haematology, plasma chemistry and PF variables. METHODS: Six healthy, mature horses were sedated with an i.v. bolus of detomidine (0.02 mg/kg bwt) and butorphanol (0.02 mg/kg bwt) and instrumented to determine the changes in cardiopulmonary function, haematology, serum chemistry and PF values during and after pneumoperitoneum with CO2 to 15 mmHg pressure for standing laparoscopy. Each horse was assigned at random to either a standing left flank exploratory laparoscopy (LFL) with CO2 pneumoperitoneum or sham procedure (SLFL) without insufflation, and instrumented for measurement of cardiopulmonary variables. Each horse underwent a second procedure in crossover fashion one month later so that all 6 horses had both an LFL and SLFL performed. Cardiopulmonary variables and blood gas analyses were obtained 5 mins after sedation and every 15 mins during 60 mins baseline (BL), insufflation (15 mmHg) and desufflation. Haematology, serum chemistry analysis and PF analysis were performed at BL, insufflation and desufflation, and 24 h after the conclusion of each procedure. RESULTS: Significant decreases in heart rate, cardiac output and cardiac index and significant increases in mean right atrial pressure, systemic vascular resistance and pulmonary vascular resistance were recorded immediately after and during sedation in both groups of horses. Pneumoperitoneum with CO2 at 15 mmHg had no significant effect on cardiopulmonary function during surgery. There were no significant differences in blood gas, haematology or plasma chemistry values within or between groups at any time interval during the study. There was a significant increase in the PF total nucleated cell count 24 h following LFL compared to baseline values for LFL or SLFL at 24 h. There were no differences in PF protein concentrations within or between groups at any time interval. CONCLUSIONS: Pneumoperitoneum with CO2 during standing laparoscopy in healthy horses does not cause adverse alterations in cardiopulmonary, haematology or plasma chemistry variables, but does induce a mild inflammatory response within the peritoneal cavity. POTENTIAL RELEVANCE: High pressure (15 mmHg) pneumoperitoneum in standing sedated mature horses for laparoscopic surgery can be performed safely without any short-term or cumulative adverse effects on haemodynamic or cardiopulmonary function. PMID- 12755433 TI - Validation and clinical utility of a novel immunoradiometric assay exclusively for biologically active whole parathyroid hormone in the horse. AB - REASONS FOR PERFORMING STUDY: Parathyroid hormone (PTH) plays a critical role in the regulation of mineral metabolism in mammals. Until recently, the standard method for PTH measurement has been the 2nd generation intact-PTH (I-PTH) assay. Current evidence indicates that the I-PTH assay binds to the PTH molecule and to an inactive N-terminally truncated PTH fragment that tends to accumulate in the blood of uraemic patients. Therefore, a new 3rd generation PTH assay that detects only the whole PTH molecule (W-PTH; cyclase-activating PTH [CAP]) has been developed. OBJECTIVES: To validate this more specific W-PTH assay for measurement of equine PTH and evaluate its clinical utility. METHODS: W-PTH and I-PTH were measured in plasma samples from normal horses (adults and foals) and horses with nutritional secondary hyperparathyroidism (N2HPT) and with chronic renal failure (CRF). Replicate measurements and dilutional paralellism were used for assay validation. Changes in blood ionized calcium were induced by EDTA and CaCl2 administration. RESULTS: Performance of the W-PTH assay (accuracy, sensitivity, specificity and ability to detect changes in PTH in response to changes in calcium) was similar to that of the I-PTH assay. Surprisingly, the relative W-PTH concentration in normal horses and foals was higher than the relative I-PTH concentration. W-PTH values remained higher than I-PTH during acute hypo- and hypercalcaemia. An increase in both W-PTH and I-PTH concentrations was found in horses with N2HPT. In horses with CRF, W-PTH and I-PTH values were very low and no increase in I-PTH was observed. CONCLUSIONS: The W-PTH assay can be used for measurement of equine PTH. POTENTIAL RELEVANCE: The use of W-PTH assay is likely to improve the diagnosis of mineral metabolism in horses. PMID- 12755434 TI - Effects of propylthiouracil and bromocryptine on serum concentrations of thyrotrophin and thyroid hormones in normal female horses. AB - REASONS FOR PERFORMING STUDY: There exists a need for better diagnostic tests to characterise thyroid disease in horses. Currently available diagnostic tests fail to differentiate between thyroid gland disorders and thyroid abnormalities resulting from pituitary or hypothalamic problems. OBJECTIVES: To evaluate the effects of treatment with propylthiouracil (PTU) and bromocryptine (BROM) on serum concentrations of triiodothyronine (T3), thyroxine (T4), reverse T3 (rT3) and equine thyroid-stimulating hormone (e-TSH, thyrotrophin) in mature horses. METHODS: Healthy mature horses were treated using either PTU or BROM for 28 days. The effect of treatment on the thyroid axis was assessed by measuring T3, T4, rT3 and e-TSH before and at +14 and +28 days. The effect of PTU and BROM on the response of T3, T4, rT3 and e-TSH to thyrotrophin-release hormone (TRH) administration was also assessed before and at +14 and +28 days of treatment. RESULTS: Treatment with PTU led to a significant reduction in serum concentrations of T3, T4 and rT3 on Day 28 and increase of e-TSH on Day 28 (P < 0.05). Treatment with BROM did not cause any measurable effect on serum concentrations of T3, T4, rT3 or e-TSH. The percentage increment by which serum concentration of T4, T3 and e-TSH increased following stimulation with TRH was decreased by treatment with PTU for 28 days (P < 0.05) but were not affected by treatment with BROM for 28 days. CONCLUSIONS: These results suggest that 1) treatment with PTU may be used in horses as a model of primary hypothyroidism; 2) the use of BROM as a model of secondary hypothyroidism in horses is not supported; and 3) e-TSH assay deserves further investigation for the clinical diagnosis of thyroid axis dysfunction in horses. POTENTIAL RELEVANCE: Propylthiouracil effectively causes primary hypothyroidism. There is substantial variability between horses with respect to their sensitivity to this substance when administered orally. Further studies pertaining to the characterisation of equine thyroid disorders are warranted and the use of both PTU for the experimental induction of primary hypothyroidism and e-TSH for the diagnostic characterisation of thyroid disorders in horses should be considered. PMID- 12755435 TI - Recombinant equine growth hormone administration: effects on synovial fluid biomarkers and cartilage metabolism in horses. AB - REASONS FOR PERFORMING STUDY: Recombinant equine growth hormone (reGH) has recently been evaluated for effects on body condition and wound healing. It has the potential to influence articular cartilage via stimulation of IGF-1. OBJECTIVES: To investigate effects of administration on synovial joint metabolism. METHODS: Six mature horses were given 20 microg/kg bwt reGH daily for 8 weeks by i.m. injection. Three control horses were injected with sterile water. Serum and synovial fluid samples were collected at 6, 8, 11 and 16 weeks for GH and IGF-1 assays. Articular cartilage harvested at week 16 was evaluated by Western analysis using monoclonal antibodies BC-13, BC-4, 8-A-4 and CH-3. RESULTS: Concentrations of IGF-1 in serum and synovial fluid were significantly elevated (P < 0.05) at 6 and 8 weeks in the reGH group. Glycosaminoglycan concentrations in synovial fluid were significantly less than controls at these time points, suggesting that reGH may modulate proteoglycan metabolism in articular cartilage. In the reGH group, there were not any alterations in synovial fluid content of 3B3(-) epitope or aggrecan metabolite, or in aggrecan or link protein catabolites retained within cartilage, that might be expected with development of osteoarthritis. CONCLUSIONS: Intramuscular administration of reGH may be a more efficient means of delivery of IGF-1 to joints for cartilage resurfacing initiatives. POTENTIAL RELEVANCE: We found no alterations in cartilage metabolism indicative of development of osteoarthritis. PMID- 12755436 TI - Medetomidine-ketamine anaesthesia induction followed by medetomidine-propofol in ponies: infusion rates and cardiopulmonary side effects. AB - REASONS FOR PERFORMING STUDY: To search for long-term total i.v. anaesthesia techniques as a potential alternative to inhalation anaesthesia. OBJECTIVES: To determine cardiopulmonary effects and anaesthesia quality of medetomidine ketamine anaesthesia induction followed by 4 h of medetomidine-propofol anaesthesia in 6 ponies. METHODS: Sedation consisted of 7 microg/kg bwt medetomidine i.v. followed after 10 min by 2 mg/kg bwt i.v. ketamine. Anaesthesia was maintained for 4 h with 3.5 microg/kg bwt/h medetomidine and propofol at minimum infusion dose rates determined by application of supramaximal electrical pain stimuli. Ventilation was spontaneous (F(I)O2 > 0.9). Cardiopulmonary measurements were always taken before electrical stimulation, 15 mins after anaesthesia induction and at 25 min intervals. RESULTS: Anaesthesia induction was excellent and movements after pain stimuli were subsequently gentle. Mean propofol infusion rates were 0.89-0.1 mg/kg bwt/min. No changes in cardiopulmonary variables occured over time. Range of mean values recorded was: respiratory rate 13.0-15.8 breaths/min; PaO2 29.1-37.9 kPa; PaCO2 6.2-6.9 kPa; heart rate 31.2-40.8 beats/min; mean arterial pressure 90.0-120.8 mmHg; cardiac index 44.1-59.8 ml/kg bwt/min; mean pulmonary arterial pressure 11.8-16.4 mmHg. Recovery to standing was an average of 31.1 mins and ponies stood within one or 2 attempts. CONCLUSIONS: In this paper, ketamine anaesthesia induction avoided the problems encountered previously with propofol. Cardiovascular function was remarkably stable. Hypoxaemia did not occur but, despite F(I)O2 of > 0.9, minimal PaO2 in one pony after 4 h anaesthesia was 8.5 kPa. POTENTIAL RELEVANCE: The described regime might offer a good, practicable alternative to inhalation anaesthesia and has potential for reducing the fatality rate in horses. PMID- 12755437 TI - Are the material properties and matrix composition of equine flexor and extensor tendons determined by their functions? AB - REASONS FOR PERFORMING STUDY: Injury to the superficial digital flexor tendon (SDFT) is common in competition horses. The SDFT contributes to locomotory efficiency by storing energy; such tendons have low safety margins. Tendons which merely position the limb, including the opposing common digital extensor tendon (CDET), are rarely injured. The current failure of strategies to prevent or effectively treat injury to the SDFT indicates the importance of understanding how it differs from tendons which are not injury-prone. HYPOTHESIS: That the structural and material properties and matrix composition of the SDFT and CDET differ, reflecting their specific functional requirements in vivo. METHODS: Forelimb tendons were harvested from 26 mature horses and loaded to failure prior to matrix composition analysis of specimens. RESULTS: The SDFT had a significantly higher cross-sectional area, structural stiffness, failure load and failure strain and a lower elastic modulus than the CDET (P < 0.0001). CONCLUSIONS: The SDFT has conflicting requirements for strength and elasticity; although as a whole it is a stiffer structure than the CDET, differences in the matrix molecular composition including water and total sulphated glycosaminoglycan contents allow it to remain more elastic as a material. POTENTIAL RELEVANCE: Further information on how the two tendons attain these different properties may be of use in the development of prevention and treatment strategies for SDFT rupture. PMID- 12755438 TI - Cutaneous leishmaniosis in three horses in Spain. PMID- 12755439 TI - Bilateral optic disc colobomas in a Quarter Horse filly. PMID- 12755440 TI - Ultrasonographic abnormalities detected in the sacroiliac area in twenty cases of upper hindlimb lameness. PMID- 12755441 TI - Long-term SO2 dispersion modeling over a coastal region. AB - Air dispersion modeling over coastal regions has proven to be a remarkable challenge in the field of air quality. Many conventional plume dispersion models, such as ISC2 and HYSPLIT, are unable to model such dispersion with the precision that is necessary to accurately predict ground-level concentrations in coastal areas. Considering this, the present work was carried out with two primary objectives: i) to evaluate the effectiveness of currently available mathematical models in predicting plume dispersion over a coastal region and ii) to study the impact of sulfur dioxide emissions from a petroleum refinery over a different community located in the adjacent area. This study demonstrates that CALPUFF predictions are more reliable compared to those of the other models studied, however the operation of CALPUFF is highly data intensive and in many instances, it is difficult to obtain all required input data. This is a particular problem for regions outside ofthe United States of America where sufficient data is difficult to obtain. In addition, the study concluded that the predicted annual average SO2 concentrations in the nearby communities are well within regulatory limits. PMID- 12755442 TI - Kinetic studies of attachment and detachment of microbial cells from soil. AB - A mathematical model was developed to describe the kinetics of cell attachment and detachment from soil. Soil-column experiments were performed to evaluate the model parameters. Pseudomonas putida G7 capable of degrading naphthalene was used as a model microorganism. A sediment sample taken from an uncontaminated area near a coal tar waste site in upstate New York, USA was used as a test soil. The kinetics of cell attachment and detachment from the model soil could be described by the developed first-order model. The equilibrium constant of attachment (11.4 ml g(-1)), the rate coefficient of cell attachment (0.299 ml g(-1) min(-1) and the rate coefficient of cell detachment (0.0263 min(-1)) were determined from the soil-column experiment. The equilibrium constant of attachment determined in this study (11.4 ml g(-1)) was within the range of those reported in the literature for bacterial attachment to soil (0.55 to 12.6 ml g(-1)). The kinetic model succesfully predicted the data of batch experiment for cell attachment and detachment soil. PMID- 12755443 TI - Observations of the impacts of some landfill leachates with clays. AB - Compacted clay liners are common, major, components of landfill leachate (fluid) containment systems. This is sensible, but knowledge and understanding of the longterm performance and behaviour of day mineral/landfill leachate systems remain very limited. The authors studied the reactions of day soil with leachate simulants related to three different climates and waste cultures. The day came from a Tertiary sequence near Melbourne, Australia, a type in common use locally for landfill engineering. X-ray diffraction was used to observe mineralogical change in 3 mm clay plugs caused by reactions with the leachate simulants. Changes in hydraulic conductivity were also measured. The results show both that the different leachates have distinct effects on the clay minerals, and that the leachate/day reactions have direct measurable and distinct impacts on hydraulic conductivity. The laboratory studies were completed at the University of Melbourne. The X-ray diffraction work was completed at The Natural History Museum in London. The experimental results are discussed here and indications given of some potential implications. PMID- 12755444 TI - Development of flexible fibre biofilm reactor for treatment of food processing wastewater. AB - Biological treatment methods are usually effective and commonly used in the treatment of organic wastewater. However, there are a number of problems for conventional biological treatment methods in treating wastewater from industries such as food processing. For example, microorganisms in the aeration tank in an activated sludge process cannot survive a continuous series of shock loads and the process could become operationally unstable. The associated problem of sludge bulking often occurs when the organic loading to the treatment process is high. In this paper, a new flexible fibre biofilm reactor was developed for the treatment of wastewater from fruit and vegetable processing plants. Experiments were carried out to evaluate the performance of the treatment process. Acclimatisation characteristics of the treatment process were also evaluated. The removal efficiencies for COD and BOD5 for different influent organic strengths evaluated. Results indicated that over 90% COD removal and 95% BOD5 removal could be achieved. The performance parameters were also compared with a conventional activated sludge process under similar conditions, operated in parallel. It was found that the biofilm reactor exhibited a number of advantages over the conventional reactor. These include (a) high organic loading rate, (b) long sludge retention times and low sludge discharge rate in the settling tank (about 10%), (c) elimination of the sludge recycle stream, and (d) no sludge bulking problem at high organic loading rates. Therefore, the flexible fibre biofilm reactor can provide a more efficient and cost effective treatment for wastewater from food processing industries. PMID- 12755445 TI - Acetate synthesis from H2/CO2 in simulated and actual landfill samples. AB - To investigate the characteristics of biological decomposition in a municipal solid waste landfill, laboratory simulated samples and full-scale actual samples excavated from landfills in Taiwan were used. Nitrogen gas was produced continuously in the simulated samples. Although nitrate or nitrite could not be detected in the leachate, this did not indicate that nitrate or nitrite was not available in the solid waste. Landfills both produce and consume H2 under anaerobic conditions. In the suspension experiment, where H2/CO2 were used as substrate, significant acetate concentration accumulated concomitantly along with hydrogen consumption. Relatively low production of methane was observed in both the simulated sample and actual samples. Collectively, acetate synthesis was the dominant reaction in the simulated sample and actual samples for hydrogen consumption. This study was the first paper reporting that the homoacetogenesis was the significant process in the landfill for hydrogen utilization, which was validated by the consumption of 4.01-5.43 moles of hydrogen per mole acetate formed. The significant acetate accumulation indicated that the homoacetogens outcompeted the hydrogentrophic methanogens in the landfill samples. Metabolically versatile homoacetogens can use many more substrates simultaneously than their competitor which is bound only to a very few substrates. Themetabolic versatility of homoacetogens could be the main reason for their success in the landfill PMID- 12755446 TI - Evaluation of organics leaching from solidified/stabilized hazardous wastes using a powder reactivated carbon additive. AB - Cement based stabilization/solidification systems have been widely used to minimize leaching of contaminants from hazardous wastes, particularly for stabilization of wastes that contain heavy metals or other inorganic materials. However, few researchers have demonstrated that stabilization/solidification systems can adequately trap and retain organics present in these wastes. Identification of additives that can be used with the inorganic binder to retain organics has attracted much interest. Activated carbon has been found to be the best additive for immobilizing organic contaminants, but it is generally too expensive for routine use. The use of powdered reactivated carbon was studied in this research to prevent organics from leaching from solidified/stabilized waste forms because it is effective and more economically competitive. The Shrinking Unreacted Core leach test method was employed in this study. Stabilization/solidification waste forms were prepared using five organic compounds--phenol, 2-chlorophenol, chlorobenzene, aniline and Methyl Ethyl Ketone (MEK)--that were spiked into the waste forms at two concentration levels each. Reactivated carbon amounts of 0% (as control), 1% and 2% (w/w) were added into each waste form batch to determine how effectively the reactivated carbon can immobilize those organics and the optimum dose of reactivated carbon addition required. Leachate samples were analyzed using gas chromatography. Results show that 1% reactivated carbon addition is generally enough to reduce leaching of all of the organic contaminants by more than 70%, while a 2% addition reduces leaching for most of the organics to less than 1%. PMID- 12755447 TI - Application of a low density support material as an alternative to prevent clogging in a three-phase fluidized-bed reactor. AB - A low density support material was evaluated to determine its effect on biofilm growth dynamics when treating synthetic wastewater in a three-phase fluidized-bed reactor. After two 40-day experimental runs, the results showed significant microbial activity from the very beginning of the study. The highest biofilm growth was observed during the first 10 days of operation. Biofilm kept gradually growing after the 11th day and reached a steady-state at day 21, defined by a consistent biomass attached to the particles. As an indicator of biofilm detachment effluent suspended biomass decreased as biofilm attachment increased, tending also to stabilize around day 21. During the first 10 days, chemical oxygen demand removal averaged 100 mg l(-1) representing 48% of the influent chemical oxygen demand concentration. The highest chemical oxygen demand removal efficiency (78%) was achieved between days 23 and 29, when influent chemical oxygen demand was 280 mg l(-1). The support material evaluated proved to be effective at allowing attachment of microorganisms. Due to its low density, bed fluidization was achieved under recirculation rates lower than those required to fluidize beds of higher density. The physical properties of the proposed support material allowed an appropriate equilibrium to be achieved between biomass attachment and detachment, eliminating the need for bed backwashing. PMID- 12755448 TI - Effects of umbrella palms and wastewater depth on wastewater treatment in a subsurface flow constructed wetland. AB - On-site subsurface flow constructed wetlands are designed to provide secondary quality effluent. Plants and wetland volume are considered in their design. There have been no studies, however, comparing wastewater treatment at different wastewater depths, and plant effects in wetlands are not completely understood. Investigations were conducted on these variables using four wetland cells 228 m wide by 4.75 m long containing 1 to 5 cm diameter river rock. Cyperus alternifolius (umbrella palms) were planted in one cell, and side-by-side comparisons were made between the planted and a control cell. Side-by-side comparisons were also made between cells with equal surface areas and different depths. At best umbrella palms improved effluent 5-day biochemical oxygen demand (BOD5) by 8%, suspended solids by 6%, and did not improve fecal coliform or P wastewater quality in July. When ambient air temperatures were < or = 12 degrees C during December, plants did not improve most treatment parameters. They did, however, significantly improve NH4+ treatment even when ambient air temperatures were as low as 8 degrees C. Increasing wastewater depth enhanced fecal coliform die-off but did not reduce effluent considered when maximum NH4+ reduction is a BOD5, suspended solids, NH4+, or P in effluent. Umbrella palms should be treatment goal and it is not necessary to reduce other parameters. Surface area was more important to wastewater treatment than depth. PMID- 12755449 TI - Phototransformation of diuron in aqueous solution by UV irradiation in the absence and in the presence of H2O2. AB - The phototransformation of diuron has been studied by photolysis at 253.7 nm at 20 degrees C, in the absence and in the presence of H2O2. Experiments were conducted in batch and in continuous-flow reactors. In the absence of H2O2, the value of the quantum yield of photolysis of diuron at 253.7 nm was found to be equal to be 0.0125 +/- 0.0005 (using a molar absorption coefficient of 16500 +/- 500 M(-1) cm (-1) at 253.7 min) and insensitive to pH in the range 2-8.5. Oxidation rates of diuron by H2O2/UV could be predicted successfully by a kinetic model including photochemical and OH*-oxidation reactions using a value of 4.6 x 10(9) M(-1) s(-1) for the rate constant of the reaction of OH* with diuron. The model was verified for the various reactors used and under a wide range of conditions in pure water (pH: 2-8, [H2O2] : 0-0.1 M) and in the presence of hydrogenocarbonate ions (0-35 mM, pH = 8.3-8.4). The contribution of the carbonate radicals to the degradation rates of diuron was found to be insignificant under our experimental conditions. PMID- 12755450 TI - Removal of some heavy metal cations from aqueous solutions by spruce sawdust. II. Adsorption-desorption through column experiments. AB - The adsorption-desorption of copper ions and a mixture of five metal ions (cadmium, copper, nickel, lead and zinc) in aqueous solutions by a spruce sawdust column was reported. Different studies about the copper binding mechanism, the efficiency of some regenerating solutions for the copper desorption, the use of column for copper adsorption-desorption by successive cycles as well as the adsorption-desorption of a multi-component solution were investigated. An ion exchange mechanism seemed to explain the removal of copper by the natural components of sawdust, i.e., calcium, magnesium and manganese accounting for 71, 13 and 5.5% respectively of the copper binding on sawdust. But there was evidence variation in the contribution of these ions during the copper adsorption in the continuous flow process. Regeneration ofthe sawdust column was described by the efficiency sequence: H+ > Ca2+ >> Na+, where protons were the most promising regenerating agents. Adsorption-desorption cycles showed that copper binding capacity of sawdust, after a decrease of 23% between cycles 1 and 2, was stabilized at 3.1 x 10(-2) meq g(-1) for the following cycles. Adsorption of a mixture of five metal ions indicated that nickel broke through first when saturation of sawdust was reached, followed by the other metal ions in the order zinc, cadmium, copper and lead. The effects of competitive ion exchange, because of affinity differences between the metal ions for sawdust, resulting in the metals having the lower affinity being displaced by those having a higher one and to overshoot the 0.2 meq l(-1) feed solution. PMID- 12755451 TI - Effects of system variables on surfactant enhanced electrokinetic removal of polycyclic aromatic hydrocarbons from clayey soils. AB - Numerous polycyclic aromatic hydrocarbon (PAH)-contaminated sites threaten public health and the environment because PAHs are commonly toxins, mutagens, and/or carcinogens. PAHs are hydrophobic and resistant to degradation; therefore, conventional remediation methods are often costly or inefficient, especially when the PAHs are present in low permeability clayey soils. Electrokinetically enhanced in-situ oil flushing is an innovative technology that has the potential to greatly increase soil-solution-contaminant interaction and remedial efficiency, even under low permeability soil conditions. Although this technique is promising, many system variables may affect remedial efficiency, such as the surfactant concentration, pH control and voltage gradient. The objective of this investigation was to evaluate the effects of these system variables. Bench-scale electrokinetic experiments were conducted using various operating conditions, which included deionized water or a 3% or a 5% surfactant concentration flushing solution. Additional tests were also conducted using the 5% surfactant concentration along with a higher pH solution or a larger voltage gradient. The experiments employing the surfactant flushing solution had a lower electroosmotic flow but exhibited greater contaminant desorption, solubilization and migration. The benefits of using a higher pH solution or a larger voltage gradient were difficult to discern because changing these process variables did not significantly improve remedial efficiency. Overall, the results indicated that the surfactant flushing solution was advantageous for treating the soil near the anode region, but contaminant migration was limited by changes in the soil and/or solution chemistry that occurred with time and/or distance from the anode. PMID- 12755452 TI - Modeling of gel barrier formation by using colloidal silica in saturated media. AB - The applicability of constructing a subsurface barrier by injecting a Colloidal Silica solution directly into the saturated medium was examined through numerical experiments. Investigations ascertain that the construction of this Colloidal Silica-based subsurface barrier in the saturated medium might be a promising containment technology. According to the results of numerical studies, gel formation takes place in the region were more than 90% of introductory concentration of the Colloidal Silica solution is maintained at a time equal to the gel-point. By using a point injection method, the radius of the solidified Colloidal Silica mass increases proportional to the respective ratios of the gel points and the injection pressure heads. With higher injection pressures and longer gel-points, a larger volume of injected Colloidal Silica solution is diluted. The effect of subsurface water flow is significant when the solution has a slower gelation rate and the spherical shape of the solidified mass is less distorted with increasing injection rate. The degree of distoration of the solidified Colloidal Silica mass is much less than that of the rest of the Colloidal Silica solution surrounding the solidified Colloidal Silica mass PMID- 12755453 TI - The usefulness of laboratory tests in the early assessment of severity of acute pancreatitis. AB - Acute pancreatitis is a disorder that affects approximately 200,000 individuals in the U.S. annually. While most cases are mild, up to 30% of patients will have a complicated course with prolonged hospitalization and significant morbidity and mortality. Early institution of several therapeutic interventions, such as enteral nutrition, prophylactic antibiotics, endoscopic retrograde cholangiopancreatography (ERCP) and intensive care monitoring, have been shown to decrease the morbidity associated with severe acute pancreatitis. However, the ability of clinicians to predict, upon presentation, which patient will have mild or severe pancreatitis has remained poor over the years, thus leading to a delay in the institution of such treatments. Researchers have focused on markers that might improve upon clinical prediction alone. While data have shown the predictive value of tools such as Ranson's and Glasgow's criteria, C-reactive protein (CRP) and the APACHE score, their application in clinical practice has been limited by a time delay of at least 48 h in the former two and by being cumbersome in the latter. Thus, our focus is to critically appraise the evidence available for various biochemical markers in their ability to distinguish mild and severe acute pancreatitis early and more accurately than currently available tools. PMID- 12755454 TI - Molecular mechanisms and regulation of iron transport. AB - Iron homeostasis is primarily maintained through regulation of its transport. This review summarizes recent discoveries in the field of iron transport that have shed light on the molecular mechanisms of dietary iron uptake, pathways for iron efflux to and between peripheral tissues, proteins implicated in organellar transport of iron (particularly the mitochondrion), and novel regulators that have been proposed to control iron assimilation. The transport of both transferrin-bound and nontransferrin-bound iron to peripheral tissues is discussed. Finally, the regulation of iron transport is also considered at the molecular level, with posttranscriptional, transcriptional, and posttranslational control mechanisms being reviewed. PMID- 12755455 TI - Acetaldehyde, microbes, and cancer of the digestive tract. AB - Excessive alcohol consumption and heavy smoking are the main risk factors of upper digestive tract cancer in industrialized countries. The association between heavy drinking and cancer appears to he particularly prominent in Asian individuals who have an inherited deficient ability to detoxify the first metabolite of ethanol oxidation, acetaldehyde. Alcohol itself is not carcinogenic. However, according to cell culture and animal experiments acetaldehyde is highly toxic, mutagenic, and carcinogenic. In addition to somatic cells, microbes representing normal human gut flora are also able to produce acetaldehyde from ethanol. After the ingestion of alcoholic beverages, this results in high local acetaldehyde concentrations in the saliva, gastric juice, and the contents of the large intestine. In addition, microbes may produce acetaldehyde endogenously without alcohol administration. This review summarizes the epidemiological, genetic, and biochemical evidence supporting the role of locally produced acetaldehyde in the pathogenesis of digestive tract cancer. Special emphasis is given to those factors that regulate local acetaldehyde concentration in the contents of the gastrointestinal tract. The new evidence presented in this review may open a microbiological approach to the pathogenesis of digestive tract cancer and may have an influence on future preventive strategies. PMID- 12755456 TI - The effects of venlafaxine on the subjective, reinforcing, and cardiovascular effects of cocaine in opioid-dependent and non-opioid-dependent humans. AB - The effects of maintenance on venlafaxine, which blocks both norepinephrine and serotonin reuptake, on the response to smoked cocaine (0, 12, 25, or 50 mg) in 7 opioid-free and 7 methadone-maintained cocaine abusers was examined during a 42 day study. Participants received venlafaxine (225 mg daily) and placebo as part of a double-blind crossover design. Cocaine significantly increased heart rate, blood pressure, cocaine choice, cocaine ratings, and ratings of positive subjective effects (e.g., "I feel high") in both groups. Venlafaxine significantly decreased the subjective effects of cocaine by 10-20% without affecting cocaine choice or cardiovascular response in both groups. Although the reduction in cocaine's effects was small, further studies using a longer venlafaxine maintenance period or a larger venlafaxine dose are warranted. PMID- 12755457 TI - Comparison between two measures of delay discounting in smokers. AB - Agreement between computer and questionnaire measures of delay discounting in smokers was compared. Correlations between measures for small, medium, or large rewards were significant. Log k values decreased as the reward delay increased, with values lower for the computer task than the questionnaire, with significant differences for small rewards. The 2 measures were related to smoking rate but not to age, gender, or obesity. The Bland-Altman test of agreement indicated large within-subject differences in k values between the 2 measures. The size of the difference between the log k values and magnitude of the log k values were positively related. Results suggest k values from the 2 measures are related but may not be used interchangeably. PMID- 12755458 TI - Delay discounting of real and hypothetical rewards. AB - The degree to which real and hypothetical rewards were discounted across delays ranging from 6 hr to 1 year was explored in a within-subjects design. An adjusting-amounts procedure was used to estimate the subjective value of real and hypothetical rewards at each delay. A hyperbolic discounting function provided a significantly better fit to individual participants' preferences than did an exponential function. No significant effect of reward type on degree of hyperbolic discounting or area under the discounting curves was detected. These findings offer some support for the validity of using hypothetical rewards to estimate discounting rates in substance-abusing and other populations, but caution is suggested because this support is gleaned from a failure to detect an effect of reward type. PMID- 12755459 TI - Diagnostic compared with abstinence outcomes of day treatment and contingency management among cocaine-dependent homeless persons. AB - Substance use disorder diagnoses were used as a treatment outcome measure in a randomized comparison of day treatment (DT) and day treatment plus contingency management (DT+) among homeless persons with primarily crack/cocaine disorders. Participants (N = 127, DT+ = 69, DT = 58, 73.2% male, 82.7% African American) were assessed at baseline and 6-month treatment completion. Binary positive and negative diagnostic outcomes for cocaine, marijuana, and alcohol were compared by treatment group. DT+ was 2.1 times more likely to have a positive treatment outcome than DT. Concordance between diagnostic change and point and continuous abstinence outcomes were found. The use of diagnostic change can be a practical addition to drug toxicology and self-report treatment outcome measures for research and practice. PMID- 12755460 TI - Aggressiveness, family history of alcoholism, and the heart rate response to alcohol intoxication. AB - Some sons of male alcoholics (SOMAs) are characterized by an increased heart rate (HR) response to alcohol intoxication, which is thought to reflect increased sensitivity to alcohol-induced reward. Such a response has also been related to increased physical aggression. However, the confounding effect of aggression in SOMAs may be obscuring the interpretation of these findings. The HR response to alcohol was therefore assessed in 4 groups: high/low aggressive SOMAs and high/low aggressive non-SOMAs. Results indicate that aggressive SOMAs had the highest intoxicated HR response and that they reported the most alcohol consumption. This suggests that in some cases the high comorbidity between alcohol misuse and aggression is related to an increased sensitivity to alcohol induced reward. PMID- 12755461 TI - Effects of alcohol on controlled and automatic memory processes. AB - The authors used the process-dissociation procedure (L. L. Jacoby, 1998) to examine the effects of alcohol on controlled and automatic influences on memory performance. Participants studied 1 of 2 word lists and subsequently were cued with word stems to recall the words from both lists. Fifty-four men were administered either a moderate dose of alcohol (0.82 g/kg) or placebo prior to studying the word list. Results indicated that alcohol decreased estimates of controlled contributions to performance on the task. In contrast, alcohol did not appear to affect automatic influences on this task. Integrated with recent findings using a different cognitive task, these data suggest that alcohol impairs performance on implicit, conceptually driven tasks but not on implicit, perceptually driven tasks. PMID- 12755462 TI - Effects of negative and positive mood phrases on priming of alcohol words in young drinkers with high and low anxiety sensitivity. AB - This study investigated whether potential emotional cues for drinking activate alcohol concepts in young drinkers. Participants were 84 university freshmen with high or low levels of anxiety sensitivity (AS). A verbal priming task measured activation (i.e., priming) of alcohol concepts (e.g., beer) by positive and negative mood phrases. Time to read alcohol target words was the dependent measure. Negative mood phrases consistently primed alcohol targets; positive mood phrases did not. Degree of negative mood priming did not differ as a function of gender or AS. Reported tendency to drink in bad moods predicted negative mood priming in women, whereas men showed negative mood priming irrespective of their reported drinking tendency. A general association between negative mood priming and severity of alcohol problems also emerged. PMID- 12755463 TI - More on axillary lymphadenectomy. PMID- 12755464 TI - Initial lymphatics--morphology and function of the endothelial cells. AB - In considering lymph formation, the function of the initial lymph sinus is usually considered a passive process. The cells, however, of the initial lymphatics hold a key position in absorbing fluid from the interstitial space. The present study using rats and guinea pigs in different days of the estrus cycle or pregnancy examined by light, scanning electron, and transmission electron microscopy suggests that the forming and closing of the open-junction formations is an active component of the lymphatic endothelium. Open-interface structures represent a further entry into initial lymphatic pathways. The existence of "endothelial cellular buds" probably act structurally to build elements of the initial lymph sinus. In short, the endothelium of the initial lymphatic sinus appears to be a structure of utmost flexibility able to respond promptly to increased amounts of lymph fluid transported from the interstitial space. PMID- 12755465 TI - Oral tolerance to dextrin mediated by specific suppressor T-cells induced in the intestinal intraepithelium and their systemic migration. AB - Antigens presented to the immune system through the oral route induce antigen specific secretory IgA and systemic unresponsiveness, termed oral tolerance (OT). We studied the induction of OT towards a diet antigen: dextrin (DEX) in rats that underwent protein deprivation and were further re-fed. Peyer's patches (PP), mesenteric lymph nodes (MLN) and spleen (Sp) cells from protein re-fed (R) rats mediated hyporesponsiveness after transfer into naive recipient rats. Low numbers of MLN T cells transferred hyporesponsiveness while higher numbers transferred an enhancement of the delayed type hypersensitivity (DTH) reaction. MLN T cells were further separated based on their ability to bind Vicia villosa (VV). MLN VV- T cells, mainly CD8+, mediated hyporesponsiveness and MLN VV+ T cells (CD45RC+ CD4- CD8- cells) abrogated the hyporesponsiveness. Moreover, Sp DEX adherent T cells were mainly CD8+. Intestinal intraepithelial lymphocytes (iIELs) mainly CD8alpha+ gamma(delta)-TCR+ cells also inhibited the DTH response to DEX after transfer. The positive DTH response to another carbohydrate (levan) indicates the specificity of the suppression to dextrin. Therefore, our data indicate that after oral administration of DEX, two different populations of T cells were generated: one found only in the MLN that mediated DTH responses and the other one capable of migrating from the intestinal intraepithelium through PP and MLN to the Sp, mediating systemic tolerance. PMID- 12755466 TI - Blue liposomes for identification of the sentinel lymph nodes in pigs. AB - Effective preoperative staining of regional lymph nodes improves their intraoperative identification and thus the selectivity of "sentinel node" lymphadenectomy. Unfortunately, aqueous Patent Blue V (PBV) often fails to provide the requisite intensity and duration of contrast (1,2). A comparative study of staining characteristics of PBV in aqueous solution with those of liposomally encapsulated PBV was carried out on 7 female pigs with an average weight of 40 kg. The liposomes, consisting of lecithin and cholesterol in a molar ratio of 3:1, were produced by an extrusion technique using membranes with thicknesses of 5 microm, 1.2 microm, 0.4 microm and 0.2 microm. In each instance, a 0.5-ml depot containing 25 +/- 0.4 mg of PBV/ml was injected into each of the four upper and lower mammary glands in aqueous solution on the left side and in a liposomal preparation on the right side. Stained lymph nodes of the groin, pelvis and neck were identified after 3, 6, 12 or 24 hours, then excised and photographed. Their PBV concentrations were measured by spectrophotometry. In all cases, the liposome preparations provided greater intensity and a longer duration than the aqueous solution. Liposomal PBV compared with aqueous PBV can therefore be recommended for better preoperative lymph node staining and identification of "sentinel" lymph nodes. PMID- 12755467 TI - Pre and postnatal exposure to endosulfan in Wistar rats. AB - The possible reproductive adverse effects of the pesticide endosulfan on male offspring rats exposed in utero and during lactation were investigated. Dams were treated orally with 0, 0.5 or 1.5 mg of endosulfan/kg 21 days prior to mating, during the mating, pregnancy and lactation. Maternal and reproductive outcome data and male sexual development landmarks (testis descent and preputial separation) were assessed. Reproductive endpoints of the male offspring were examined at adulthood: sex organ weights, daily sperm production, spermatid number, sperm transit, sperm morphology and testosterone level. No signs of maternal toxicity were detected at the dose levels tested. Sexual development landmarks were also unaffected. Moreover, with the exception of a significant increase in the relative epididymis weight seen in the group treated with the lowest dose, we have not found any statistically significant adverse effect in the reproductive endpoints investigated at adulthood. The results of the present study indicate that pre and postnatal exposure to low doses of endosulfan (0.5 and 1.5 mg/kg) do not induce significant adverse effects in the reproductive system of male offspring Wistar rats at adulthood. PMID- 12755468 TI - Structural and functional changes by Ciprofloxacin of rat submandibular gland. AB - In the present study, the effects of Ciprofloxacin (Cipro), a fluoroquinolone antibiotic, on rat submandibular gland structure and function were examined in an acute experiment. Cipro was administered intraperitoneally at various doses (20, 40 and 80 mg/kg). Pure submandibular saliva was collected intraorally by micropolyethylene tubes under anaesthesia using a dissecting microscope. After collection of saliva, submandibular glands were removed and weighed. Flow rate, amylase activity, total protein and electrolyte concentrations were measured in saliva. Concentrations of DNA and protein were measured in the gland. All doses of Cipro (20, 40, 80 mg/kg) reduced salivary flow rate. Concentrations of salivary total protein and calcium and gland DNA were reduced by all doses of Cipro. Treatment by Cipro (80 mg/kg) induced an increase in salivary sodium and potassium concentrations. Histopathological examination of glands revealed that Cipro at doses of 40 mg/kg and 80 mg/kg induces morphological changes in the glands including irregular shape of the cerous and mucous bobbles, lack of nucleus in some cells, damage of the cytoplasmic and cell walls and presence of oncocytes in secretory ducts. It is concluded that Cipro inhibits rat submandibular gland functions consistent with structural damages to the gland that might be observed as a side effect in humans. Properties of fluoroquinolones to alter intracellular cAMP and their ability to suppress DNA and protein synthesis of acinar cells might be possible reasons for observed changes. PMID- 12755469 TI - Combined efficacies of lipoic acid and meso-2,3-dimercaptosuccinic acid on lead induced erythrocyte membrane lipid peroxidation and antioxidant status in rats. AB - One of the most intriguing phenomenon observed during lead toxicity has been attributed to lead-induced oxidative stress. The combined effect of DL-alpha lipoic acid (LA) and meso-2,3-dimercaptosuccinic acid (DMSA) on lead-induced alterations in selected parameters, which are indicators of oxidative stress in erythrocytes, have been studied. Lead acetate (Pb, 0.2%) was administered in drinking water for 5 weeks to induce toxicity. LA (25 mg/ kg body weight per day i.p.) and DMSA (20 mg/kg body weight per day i.p.) were administered individually and also in combination during week 6. Clinical evidence of toxic exposure was evident from the elevated blood lead levels (BPb) along with lowered levels of haemoglobin (Hb) and haematocrit (Ht). Lead-exposed animals showed enhanced membrane lipid peroxidation (LPO) in the erythrocytes. Damage to the erythrocyte membrane was evident from the decline in the activities of the transmembrane enzymes, viz., Na+, K(+)-ATPase, Ca(2+)-ATPase and Mg(2+)-ATPase. Lead-exposed rats also suffered an onslaught on the antioxidant defence system witnessed by lowered activities of catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx) and reduced glutathione (GSH). Serum glutamic-oxoloacetic transaminase (SGOT) and serum glutamic-pyruvic transaminase (SGPT) were also elevated in lead-exposed rats. Treatment with either LA or DMSA reversed the lead induced biochemical disturbances encountered by the erythrocytes, but combined treatment with LA and DMSA was very effective in mitigating all the parameters indicative of oxidative stress. PMID- 12755470 TI - Nigella sativa (black cumin) ameliorates potassium bromate-induced early events of carcinogenesis: diminution of oxidative stress. AB - Potassium bromate (KBrO3) is a potent nephrotoxic agent. In this paper, we report the chemopreventive effect of Nigella sativa (black cumin) on KBrO3-mediated renal oxidative stress, toxicity and tumor promotion response in rats. KBrO3 (125 mg/kg body weight, intraperitoneally) enhances lipid peroxidation, gamma-glutamyl transpeptidase, hydrogen peroxide and xanthine oxidase with reduction in the activities of renal antioxidant enzymes and renal glutathione content. A marked increase in blood urea nitrogen and serum creatinine has also been observed. KBrO3 treatment also enhances ornithine decarboxylase (ODC) activity and [3H] thymidine incorporation into renal DNA. Prophylaxis of rats orally with Nigella sativa extract (50 mg/kg body weight and 100 mg/kg body weight) resulted in a significant decrease in renal microsomal lipid peroxidation (P < 0.001), gamma glutamyl transpeptidase (P < 0.001), H2O2 (P < 0.001) and xanthine oxidase (P < 0.05). There was significant recovery of renal glutathione content (P < 0.01) and antioxidant enzymes (P < 0.001). There was also reversal in the enhancement of blood urea nitrogen, serum creatinine, renal ODC activity and DNA synthesis (P < 0.001). Data suggest that Nigella sativa is a potent chemopreventive agent and may suppress KBrO3-mediated renal oxidative stress, toxicity and tumour promotion response in rats. PMID- 12755471 TI - Biochemical evidence for free radical-induced lipid peroxidation as a mechanism for subchronic toxicity of malathion in blood and liver of rats. AB - Organophosphorus compounds may induce oxidative stress leading to generation of free radicals and alterations in antioxidant and scavengers of oxygen free radicals (OFRs). The effect of subchronic exposure to malathion in the production of oxidative stress was evaluated in male Wistar rats. Administration of malathion (100, 316, 1000, 1500 ppm) for 4 weeks increased catalase (CAT), superoxide dismutase (SOD) activities as well as malondialdehyde (MDA) concentration in red blood cells (RBC) and liver. However, acetylcholinesterase (AChE) and cholinesterase (ChE) activities were decreased in these samples. The increase in RBC and liver lipid peroxidation correlated well with the inhibition in RBC AChE and liver ChE activities. Elevation of MDA concentrations and increased activities of CAT and SOD showed significant correlations in both RBC and liver samples when different doses of malathion were used. The results of the present study suggest the usefulness of RBC AChE measurement as a good biomarker in the estimation of malathion-induced oxidative stress affecting blood and liver. PMID- 12755472 TI - Investigation of biochemical and histopathological effects of Mentha piperita L. and Mentha spicata L. on kidney tissue in rats. AB - Peppermint plants have been used as a herbal medicine for many conditions, including loss of appetite, common cold, bronchitis, sinusitis, fever, nausea, vomiting and indigestion. This study is aimed at investigating the biochemical and histological effects of Mentha piperita L., growing in the Yenisar Bademli town of Isparta City, and Mentha spicata L., growing on the Anamas high plateau of Isparta City, on rat kidney tissue. Forty-eight male Wistar albino rats weighing 200-250 g were used for this study. Animals were divided into four experimental groups, each with 12 rats, as follows: control group (group I); 20 g/L M. piperita tea (group II); 20 g/L M. spicata tea (group III); 40 g/L M. spicata tea (group IV). The control group rats were given commercial drinking water (Hayat DANONESA water). The tea for the other groups was prepared daily and provided at all times to the rats during 30 days as drinking water. Plasma urea and creatinine levels were determined, and the levels of thiobarbituric acid reactive substance (TBARS) and the activities of glutathione peroxidase (GSH-Px), catalase (CAT) and superoxide dismutase (SOD) were studied in the homogenates of kidney tissue. The levels of plasma urea and creatinine were increased significantly (P < 0.0033) in groups III and IV when compared with group I. The activities of SOD and GSH-Px were decreased significantly (P < 0.0033) in group IV when compared with group I. The activities of CAT were decreased significantly in groups III and IV (P < 0.033, P < 0.0033, respectively) when compared with group I. TBARS levels were increased significantly (P < 0.0033) in groups III and IV when compared with group I. In groups II, III and IV, hydropic degeneration of tubular epithelial cells, the epithelial cells with picnotic nuclei and eosinophilic cytoplasm, tubular dilatation and enlargements in Bowman capsules were observed histologically. However, in group II histopathological changes were more slight than in groups III and IV. In group IV, in addition to these changes, extremely hydropic degeneration of tubular epithelial cells, some atrophic tubules and glomerules, and focal mononuclear cell infiltrations in the kidney tissues of the rats were observed. In conclusion, the results indicate that M. piperita does not show nephrotoxicity but M. spicata presents markedly nephrotoxic changes in rats. PMID- 12755473 TI - Liver death and regeneration in paracetamol toxicity. AB - Paracetamol overdose (POD) is a major clinical problem as the commonest cause of fulminant hepatic failure (FHF) in the UK and the USA. While the main loss of liver mass occurs following hepatocyte necrosis, hepatocyte apoptosis has also been reported to occur during paracetamol toxicity in murine liver. Hepatocyte apoptosis has not previously been identified in human liver and the significance of apoptosis in paracetamol toxicity is not known. In this study of paracetamol toxicity in human liver after POD, hepatocyte apoptosis was identified at time of liver transplantation or death and was associated with striking regenerative activity. The biological significance of apoptosis is unclear but the rates of apoptosis found (0.6%) could account for a significant loss of hepatic parenchyma. The stimulus for apoptosis is not known but it is unlikely to be induced directly by paracetamol since it is absent from serum at this time. The possibility that apoptosis may be induced by Kupffer cell activation with cytokine production is raised. Patients who develop FHF after POD have a poor prognosis, with few therapeutic options apart from liver transplantation; an understanding of the dynamics of liver regeneration and ongoing cell loss by apoptosis may allow the development of new therapies in these patients. PMID- 12755474 TI - Long-term analysis of diesel fuel consumption in a co-culture of Acinetobacter venetianus, Pseudomonas putida and Alcaligenes faecalis. AB - The dynamics of a microbial population isolated from superficial waters of Venice Lagoon and the ability to utilise diesel fuel (n-alkanes mixture C12-C28) as the sole carbon and energy source were studied in a long-term reconstruction experiment. The reconstructed microbial population consisted of three bacterial strains belonging to the species Acinetobacter venetianus, Pseudomonas putida, and Alcaligenes faecalis, which were able to oxidise n-alkanes to alkanoates, n alkanols to alkanoates, or only n-alkanoates, respectively. Three different approaches: plate counting, cell counting by epifluorescence microscopy with DAPI staining, and by fluorescence in situ hybridisation (FISH) by using a probe conjugate with fluoresceine isothiocyanate specifically targeted towards the 16S rRNA of bacteria belonging to the genus Acinetobacter were used to monitor the growth of the bacterial population. The growth of A. venetianus was stimulated by the presence of other strains, suggesting a beneficial interaction. After the first week of growth A. venetianus cells formed aggregates, as confirmed by confocal microscopy (CLSM), which allowed them to be distinguished from free cells. A relationship between cell number and measured areas (microm2) per aggregate was found. Each cell presented an average surface of 1.21 microm2. Each aggregate was formed by a cellular monolayer biofilm consisting of up to several thousands of cells. The A. venetianus aggregates increased in number and size over time, but after two weeks fragmentation events, which had a beneficial effect on the growth of P. putida and A. faecalis, occurred. PMID- 12755475 TI - Molecular taxonomy of dermatophytes and related fungi by chitin synthase 1 (CHS1) gene sequences. AB - In the present study, the nucleotide sequences of the CHS1 gene from dermatophytes and related fungi in the genera Chrysosporium, Epidermophyton, Microsporum and Trichophyton were investigated using molecular methods. About 440 bp genomic DNA fragments of the CHS1 gene from 21 species were amplified by polymerase chain reaction (PCR) and sequenced. The CHS1 nucleotide sequences of these fungi showed more than 83% similarity. The molecular taxonomy of the CHS1 gene sequences revealed that Microsporum was genetically distinct from Chrysosporium and Trichophyton, as classified by morphological characteristics. PMID- 12755476 TI - Streptomyces yatensis sp. nov., a novel bioactive streptomycete isolated from a New-Caledonian ultramafic soil. AB - The taxonomic position of an actinomycete isolated from an ultramafic soil in New Caledonia was examined using a polyphasic approach. The organism, which was designated SFOCin 76, was found to have chemical and morphological properties typical of streptomycetes and formed a distinct phyletic line in the Streptomyces violaceusniger clade of the 16S rDNA tree. It also showed a unique pattern of phenotypic properties that distinguished it from representatives of all of the validly described species classified in this clade. It is, therefore, proposed that strain SFOCin 76 be classified in the genus Streptomyces as Streptomyces yatensis sp. nov. PMID- 12755477 TI - Lactic acid bacteria: inhibition of angiotensin converting enzyme in vitro and in vivo. AB - A total of 26 strains of wild-type lactic acid bacteria, mainly belonging to Lactococcus lactis and Lactobacillus helveticus, were assayed in vitro for their ability to produce a milk fermentate with inhibitory activity towards angiotensin converting enzyme (ACE). It was clear that the test strains in this study, in general, produce inhibitory substances in varying amounts. Using a spectrophotometric assay based on amino group derivatization with ortho phthaldialdehyde as a measure of relative peptide content, it was shown that there is a significant correlation between peptide formation and ACE inhibition, indicating that peptide measurement constitutes a convenient selection method. The effect of active fermentates on in vivo ACE activity was demonstrated in normotensive rats. The pressor effect of angiotensin I (0.3 microg/kg) upon intravenous injection was significantly lower when rats were pre-fed with milks fermented using two strains of Lactobacillus helveticus. An increased response to bradykinin (10 microg/kg, intravenously injected) was observed using one of these fermented milks. It is concluded that Lactobacillus helveticus produces substances which in vivo can give rise to an inhibition of ACE. The inhibition in vivo was low compared to what can be achieved with classical ACE inhibitors. The clinical relevance of this finding is discussed. This work is the first in which an effect of fermented milk on ACE in vivo has been demonstrated, measured as decreased ability to convert angiotensin I to angiotensin II. PMID- 12755478 TI - An Azospirillum brasilense Tn5 mutant with modified stress response and impaired in flocculation. AB - The analysis of an A. brasilense Tn5 mutant shows significant phenotypic differences compared to the wild type isogenic strain. The transposon was located disrupting an open reading frame of 840 bp (ORF280) which exhibits similarity to the universal stress protein (USP) family. The USP family encompasses proteins that are expressed as a response to cell growth arrest. The mutant revealed a pleiotrophic phenotype with respect to different stress conditions. The ORF mutation results in an increased sensitivity of cells to carbon starvation and heat-shock treatment. However, the mutant strain displays a higher tolerance to oxidative stress agents. In contrast to the isogenic parent strain, colonies of the mutant are weakly stained by Congo red added to solid media and are impaired in flocculation. Scanning electron micrographs revealed that the mutant lacks part of the surface material present as a thick layer of exopolysaccharides on the surface of the wild type cells. The pleiotrophic phenotype revealed for this mutant and the similarity of the C-terminal region of ORF280 to UspA from E. coli indicates that the A. brasilense ORF280 may be a Usp-like protein. PMID- 12755479 TI - Influence of different combinations of bacteria and yeasts in voice prosthesis biofilms on air flow resistance. AB - Laryngectomized patients use silicone rubber voice prostheses to rehabilitate their voice. However, biofilm formation limits the lifetime of voice prostheses. The presence of particular combinations of bacterial and yeast strains in voice prosthesis biofilms has been suggested to be crucial for causing valve failure. In order to identify combinations of bacterial and yeast strains causative to failure of voice prostheses, the effects of various combinations of bacterial and yeast strains on air flow resistances of Groningen button voice prostheses were determined. Biofilms were grown on Groningen button voice prostheses by inoculating so-called artificial throats with various combinations of clinically relevant bacterial and yeast strains. After 3 days, all throats were perfused three times daily with 250 ml phosphate buffered saline and at the end of each day the artificial throats were filled with growth medium for half an hour. After 7 days, the air flow resistances of the prostheses were measured. These air flow resistances were expressed relative to the air flow resistances of the same prostheses prior to biofilm formation. This study shows that biofilms causing strong increases in air flow resistance (26 to 28 cm water x s/l) comprised combinations of microorganisms, involving Candida tropicalis, Staphylococcus aureus and Rothia dentocariosa. PMID- 12755480 TI - Effects of lycorine on growth and effects of L-galactonic acid-gamma-lactone on ascorbic acid biosynthesis in strains of Cryptococcus laurentii isolated from Narcissus pseudonarcissus roots and bulbs. AB - The alkaloid lycorine, which is considered to inhibit the last step in ascorbic acid biosynthesis, is produced by Narcissus pseudonarcissus. The growth of two strains (C1 and C3) of Cryptococcus laurentii isolated from root tips of N. pseudonarcissus is inhibited by lycorine, as is the in vivo production of ascorbic acid from L-galactonic acid-gamma-lactone. In contrast, C. laurentii strain C4, isolated from the lycorine-containing bracts of the bulb, was not inhibited by lycorine and did not contain ascorbic acid when cultivated with or without L-galactonic acid-gamma-lactone. PMID- 12755481 TI - An assessment of the in vitro antimicrobial effects of two antiepileptic drugs- sodium valproate and phenytoin. AB - An incidental observation led to the evaluation of the antimicrobial properties of valproate and phenytoin. In vitro inhibition of Mycobacterium smegmatis, Candida albicans, and other standard test organisms by these two antiepileptic drugs was assessed using the broth microdilution procedure. Fluorescence microscopy (Viability Stains, Molecular Probes Inc.) and cultural techniques were employed to distinguish between microbicidal and microbistatic effects. Phenytoin showed no inhibitory activity against the microbes tested. Sodium valproate, on the other hand, was selectively potent against the yeast strains in a dose dependent manner (MIC = 10-20 microg ml(-1)). In vitro activity against Mycobacterium smegmatis (70% growth inhibition by 81 microg ml(-1)) was moderate to low while Staphylococcus aureus. Pseudomonas aeruginosa and Escherichia coli were not significantly affected. Viability data from fluorescent microscopy and plate cultures correlated well with absorbance (A620nm) growth index, and showed that valproate was microbicidal against susceptible organisms. The mode of action may include blockage of calcium channels and perturbation of membrane potential. This report opens up yet another opportunity for further enquiry into the fundamental mechanisms of drug action and microbial resistance. PMID- 12755482 TI - Relation between phylogeny and physiology in some ascomycetous yeasts. AB - The question of whether yeasts with similar physiological properties are closely related has been examined using recently published phylogenetic analyses of 26S domain D1/D2 rDNA nucleotide sequences from all currently recognized ascomycetous yeasts. When apparently unique metabolic pathways are examined, some relationships between physiology and rDNA phylogeny are evident. Most Candida and Pichia species that are able to assimilate methanol as the sole carbon source are in a clade delimited by C. nanospora and C. boidinii. Exceptions are P. capsulata and P. pastoris which are phylogenetically separated from the other methanol assimilating yeasts. Yeasts subject to the petite mutation, resulting in respiratory deficiency, belong to three different clades, viz, a Saccharomyces clade delimited by S. cerevisiae and S. rosinii, the Dekkera/Brettanomyces clade, and some Schizosaccharomyces species ('Archiascomycete' clade). However, petite mutants were also found in Zygosaccharomyces fermentati and some other more distantly related species. Yeasts able to assimilate n-hexadecane, uric acid or amines as sole carbon source are broadly distributed over the ascomycetous phylogenetic tree. However, species that assimilate adenine as sole carbon source are closely related. Most of these species also assimilated glycine, uric acid, n hexadecane, putrescine and branched-chain aliphatic compounds such as isobutanol, leucine and isoleucine. Among the Saccharomycetales, species utilizing all or the great majority of these eight compounds are in the Stephanoascus/Arxula/Blastobotrys clade. Candida blankii, which is distantly related to this clade, proved to be an exception and assimilated six of eight of these compounds. PMID- 12755483 TI - Gordonia sinesedis sp. nov., a novel soil isolate. AB - The taxonomic position of an actinomycete isolated from soil was evaluated using a polyphasic approach. The organism, strain J72, was found to have chemical and morphological properties consistent with its assignment to the genus Gordonia. A nearly complete 16S rDNA sequence of the strain was determined by direct sequencing of the amplified gene. The tested strain formed a distinct phylogenetic line within the evolutionary radiation occupied by the genus Gordonia and was most closely related to G. polyisoprenivorans DSM 44302T. The phenotypic profile of strain 372 readily distinguishes it from representatives of the validly described species of Gordonia. The combined genotypic and phenotypic data show that strain J72 merits recognition as a new species of Gordonia. The name proposed for the new species is Gordonia sinesedis; the type strain is J72T (= DSM 44455T = NCIMB 13802T). PMID- 12755484 TI - Phylogenetic analysis of Antrodia and related taxa based on partial mitochondrial SSU rDNA sequences. AB - Sequences of mitochondrial SSU rDNA were obtained from six species of Antrodia and related fungal taxa to reveal their phylogenetic relationships. Phylogenetic analysis showed that the species of Antrodia did not cluster into a single clade. Brown rot fungi were separated into two main groups through which Antrodia was dispersed. Antrodia sinuosa, A. serialis, A. heteromorpha and A. malicola clustered with Perenniporia, Fomitopsis, Piptoporus, Daedalea and Melanoporia within one group of brown rot fungi, while A. carbonica and A. vaillantii clustered with Oligoporus, Gloeophyllum and Auriporia within the other group of brown rot fungi, indicating that Antrodia is a heterogeneous genus and that brown rot fungi have evolved convergently. PMID- 12755485 TI - The costs and benefits of killer toxin production by the yeast Pichia kluyveri. AB - Numerous yeast species in many genera are able to produce and excrete extracellular toxic proteins (mycocins) that can kill other specific sensitive yeasts. Natural distributions of killer yeasts suggest that they may be important in maintaining community composition and provide a benefit to the toxin producing cells. The fact that not all yeasts are killers and that polymorphisms exist within some killer species suggests there may be a cost associated with killer toxin production. This study focuses on the costs and benefits associated with toxin production by the yeast Pichia kluyveri. Strains differing in their ability to kill were obtained by tetrad dissection. One parent strain produced spores that exhibited a trade-off between killing ability and intrinsic growth rate. A killer clone from this strain was able to maintain a higher proportion of cells than a non-killer when grown with the same sensitive yeast under laboratory simulated natural conditions. On the other hand, when grown with a yeast not sensitive to Pichia kluyveri toxin, the non-killer maintained a higher proportion of the total community than did the killer clone. The data support the hypothesis that there are both costs and benefits to producing killer toxin, and based on this, selection may favor different phenotypes in different conditions. PMID- 12755486 TI - Copper resistance in Desulfovibrio strain R2. AB - A sulfate-reducing bacterium, designated as strain R2, was isolated from wastewater of a ball-bearing manufacturing facility in Tomsk, Western Siberia. This isolate was resistant up to 800 mg Cu/l in the growth medium. By comparison, Cu-resistance of reference cultures of sulfate-reducing bacteria ranged from 50 to 75 mg Cu/l. Growth experiments with strain R2 showed that Cu was an essential trace element and, on one hand, enhanced growth at concentrations up to 10 mg/l but, on the other hand, the growth rate decreased and lag-period extended at copper concentrations of >50 mg/l. Phenotypic characteristics and a 1078 bp nucleotide sequence of the 16S rDNA placed strain R2 within the genus Desulfovibrio. Desulfovibrio R2 carried at least one plasmid of approximately of 23.1 kbp. A 636 bp fragment of the pcoR gene of the pco operon that encodes Cu resistance was amplified by PCR from plasmid DNA of strain R2. The pco genes are involved in Cu-resistance in some enteric and aerobic soil bacteria. Desulfovibrio R2 is a prospective strain for bioremediation purposes and for developing a homologous system for transformation of Cu-resistance in sulfate reducing bacteria. PMID- 12755487 TI - Identifying and validating causal genetic alterations in human breast cancer. AB - An important mechanism for the activation of proto-oncogenes in human breast and other cancers is gene amplification, which results in gene overexpression at both the message and the protein levels. Recent studies have demonstrated that oncogenes rarely if ever become amplified in isolation, but rather are present on large amplicons that contain multiple genes. More detailed analysis of these amplicons has revealed the presence of many candidate breast cancer oncogenes. The broad goal of this issue of Breast Cancer Research and Treatment is to review the current state of our understanding of the causal role of defined genetic alterations that occur in human breast cancers, and to discuss the case for the mechanistic significance of several candidate oncogenes. As will be seen, these studies have revealed a remarkable genetic complexity and heterogeneity in human breast cancer that must be dissected in order to improve our mechanistic understanding of disease progression, and to develop effective new drugs against relevant molecular targets. PMID- 12755488 TI - Profiling breast cancer by array CGH. AB - Breast tumors display a wide variety of genomic alterations. This review focuses on DNA copy number variations in these tumors as measured by the recently developed microarray-based form of comparative genomic hybridization. The capabilities of this new technology are reviewed. Initial applications of array CGH to the analysis of breast cancer, and the mechanisms by which the particular types of copy number changes might arise are discussed. PMID- 12755489 TI - HER-2/neu and topoisomerase IIalpha in breast cancer. AB - In breast cancer, the predominant genetic mechanism for oncogene activation is through an amplification of a gene. The HER-2 (also known as ErbB2/c-erbB2/HER 2/neu) oncogene is the most frequently amplified oncogene in breast cancer, and its overexpression is associated with poor clinical outcome. In addition to its important role in breast cancer growth and progression, HER-2 is also a target for a new form of chemotherapy. Breast cancer patients have been treated with considerable success since 1998 with trastuzumab, a recombinant antibody designed to block signaling through HER-2 receptor. HER-2 has also been implicated in altering the chemosensitivity of breast cancer cells to different forms of conventional cytotoxic chemotherapy, particularly of topoII-inhibitors (e.g., anthracyclines). Topoisomerase IIalpha gene is located just by the HER-2 oncogene at the chromosome 17q12-q21 and is amplified or deleted in almost 90% of the HER 2 amplified primary breast tumors. Recent data suggests that amplification and deletion of topoisomerase IIalpha may account for both relative chemosensitivity and resistance to anthracycline therapy, depending on the specific genetic defect at the topoIIalpha locus. Expanding our understanding of HER-2 amplification also changes its role in the pathogenesis of breast cancer. HER-2 is an oncogene that clearly can drive tumor induction and growth and is also a target for a new kind of chemotherapy, but its function as a marker for chemoselection may be due to associated genetic changes, of which topoisomerase IIalpha is a good example. Moreover, despite potential evidence that genes other than HER-2, such as topoisomerase IIalpha, may be more important predictors of therapeutic response in breast cancer, HER-2 status still has a very significant role in therapeutic selection, mainly as the major criterion for administering trastuzumab in treating breast cancer. Thus, the clinical and therapeutic importance of the HER 2 and topoisomerase IIalpha status to breast cancer management should only increase in the next few years. PMID- 12755490 TI - The 17q23 amplicon and breast cancer. AB - A novel region of amplification in breast tumors was recently identified on chromosome 17q23. Extensive mapping of the amplicon by Southern blotting and fluorescence in situ hybridization (FISH) in breast cancer cell lines determined that the amplicon can be up to 4 Mbp in size and may contain 50 genes. Copy number analysis at 50-75 kb resolution in breast cancer cell lines and breast tumors identified several independently amplified regions within the amplicon, suggesting that a number of genes are selected for amplification because they independently contribute to tumor formation and progression. Support for this hypothesis comes from studies demonstrating that many of the amplified genes are over-expressed in breast cancer cell lines and tumors, and that the RPS6KB1, TBX2, and PPM1D genes from the region, that are amplified and over-expressed in breast tumors and cell lines, contribute to tumor formation and/or tumor progression. In this review we summarize the structural studies of the amplicon that have been carried out, we outline the evidence implicating the RPS6KB1, TBX2, and PPM1D genes as oncogenes, and we describe some of the other candidate oncogenes from the region. PMID- 12755491 TI - Cyclin D1, EMS1 and 11q13 amplification in breast cancer. AB - Chromosome locus 11q13 is frequently amplified in a number of human cancers including carcinoma of the breast where up to 15% carry this chromosomal abnormality. Originally 11q13 amplification was thought to involve a single amplicon spanning many megabases, but more recent data have identified four core regions within 11q13 that can be amplified independently or together in different combinations. Although the region harbors several genes with known or suspected oncogenic potential, the complex structure of the amplicons and the fact that 11q13 is gene-rich have made definitive identification of specific genes that contribute to the genesis and progression of breast cancer a difficult and continuing process. To date CCND1, encoding the cell cycle regulatory gene cyclin D1, and EMS1, encoding the filamentous actin binding protein and c-Src substrate cortactin, are the favored candidates responsible for the emergence of two of the four amplification cores. PMID- 12755492 TI - Genome amplification of chromosome 20 in breast cancer. AB - Recurrent gain and amplification of the long arm of chromosome 20 (20q) has been observed in a wide variety of cancers. This suggests that a gene or genes encoded on 20q play important roles in contributing to the cancer phenotype when overexpressed. In the quest to discover cancer genes, this region of the genome has been exhaustively studied, and the results demonstrate remarkable complexity. Multiple regions of low and high-level 20q copy number gain correlate with poor clinical prognosis and appear to contribute to the cancer phenotype, especially aspects of immortalization, genome instability, apoptosis, and increased proliferation. Gene discovery efforts have revealed a number of interesting candidate genes on chromosome 20 that may contribute to oncogenic progression. The study of 20q serves as a model for positional cloning enthusiasts, demonstrating the path typically taken when moving from initial discovery of an important genomic abnormality to identification of genes likely to be significant players in disease progression. This review will summarize approximately a decade of study on 20q and is structured as moving from an introduction to the techniques used in 20q analyses, to the details of 20q genomic complexity and its involvement with cancer, and finally to a detailed gene-specific look at this region. PMID- 12755493 TI - Genetic and cytogenetic analyses of breast cancer yield different perspectives of a complex disease. AB - Genomic instability in breast cancer results in low-level changes in DNA copy number, a significant but poorly understood mechanism underlying the genetic heterogeneity of this disorder. Two different approaches, loss of heterozygosity (LOH) and comparative genomic hybridization (CGH), have been used to probe the genetics of breast cancer evolution. LOH is a locus specific method that detects the variation in the parental origin of DNA, but is not quantitative. CGH provides a genome-wide accounting of the magnitude of DNA copy number changes, but not parental origin. Both methods have identified complex and heterogeneous patterns of DNA losses, duplications, and amplifications during breast cancer evolution. LOH and CGH technologies interrogate very distinct mechanisms driving breast tumor evolution, yet are seldom used in parallel to profile specimens. Thus, the relative significance of genetic versus numerical variations of DNA in breast cancer evolution remains undefined. This review will attempt to summarize some of the successes of these investigations, highlight some complex and confounding observations emerging from these studies, and discuss the potential of these studies to improve our understanding of breast cancer biology and treatment. PMID- 12755494 TI - Globulins enhance in vitro iron but not zinc dialysability: a study on six legume species. AB - The study was addressed to evaluate the in vitro iron and zinc dialysability from the globulin fraction of six legumes. Five legume species including white bean, mottled bean (Taylor bean), chickpea, lentil, lupin, and a modified mottled bean variety, selected by back-crossing to obtain seeds with globulins composed by G1 fraction only, were used. Globulins (G1 + G2) were extracted from the seeds and analysed for their in vitro iron and zinc dialysability. The highest globulin concentration was detected in lentil (89%). The percentage of globulins in the modified variety of Taylor bean (G1 only) was higher than that of the commercial variety (G1 + G2). The highest concentration of iron was found in Taylor bean globulins. The modified variety of Taylor bean contained 2.6-fold higher iron concentration than the whole seed, and the commercial variety had 1.8-fold higher iron only. The highest zinc concentration was found in lentil globulins. Also iron dialysability from globulins was markedly higher than that of the respective whole seed. The highest value of iron dialysability was found in lentil (10.8%). Zinc dialysability was generally high (above 20%), but no significant differences between whole seed and globulins were detected. The results showed that globulins enhanced iron but not zinc dialysability. Lupin and the modified variety of Taylor bean showed a different behaviour in terms of mineral dialysability compared to the other legumes. The amino acid composition of the digestion products of whole seeds and globulins failed to evidence any direct influence on iron and zinc availability. PMID- 12755495 TI - An evaluation of the influence of therapeutic interventions on serum trace element levels in groups of patients. AB - The aim of the study was to propose a method of large amount data evaluation. A new graphical method for data evaluation was suggested: the data were ranked according to the initial values and both the initial values and final values were intersected by polynomial curves. This method was used in the following situations: 1. Serum levels of Mg and Zn were measured just before and after hemodialysis (HD) in 87 patients in chronic renal failure. 2. Mg levels in serum, red blood cells and urine were estimated in 20 patients before and after administration of a Mg containing drug. Three basic graphic forms of curves were established: 1. Significant decrease of serum Mg levels during HD resulted in two uncrossed lines, the initial-values line being higher than terminal-values one (the higher the initial level the more pronounced was its decrease during HD). 2. Balanced effect of HD on the serum levels of Zn (low values increased, high levels decreased) represented two crossing-lines. 3. Significant increase of urine Mg in patients supplemented by Mg demonstrated two uncrossed lines. The position of initial-values curve was lower than the terminal-values one. The proposed graphical method of the evaluation of large amounts of data is simple and enables a quick orientation in the assessment of the effects of therapeutic interventions (trace elements, drugs and other relevant substances). PMID- 12755496 TI - Sex, age, geographical location, smoking, and alcohol consumption influence serum selenium concentrations in the USA: third National Health and Nutrition Examination Survey, 1988-1994. AB - Selenium has been reported to reduce the risk for heart diseases and cancer. We examined the association of sex, age, geographical location, serum cotinine concentrations, a measure of smoking intensity, and alcohol consumption with serum selenium concentrations in the third National Health and Nutrition Examination Survey (NHANES III), 1988-1994. Study sample consisted of 14,619 apparently healthy individuals (male: 7,102; female: 7,517) aged 14 to >90 years. Sex, age, geographical location, serum cotinine concentrations, and alcohol consumption significantly influenced serum selenium concentrations (P < 0.05). The mean (+/- standard error) serum selenium concentration in men (124.5 +/- 0.20 ng/mL) was significantly higher than in women (122.0 +/- 0.20 ng/mL) (P < 0.0001). Men in the 31-50 y age group had the highest mean serum selenium concentration. In the same age group, women had the lowest mean serum selenium concentration. In both sexes, participants living in the Midwest and West had significantly higher serum selenium concentrations than those living in South and Northeast geographical locations. Serum cotinine was negatively associated with serum selenium concentrations in both men (beta = -0.0108; P < 0.0001 for trend) and women (beta = -0.0097; P < 0.0001 for trend). Alcohol consumption is positively associated with serum selenium in women (beta = 0.0462; P = 0.0044 for trend) but not in men (beta = 0.0015; P = 0.8496 for trend). Although, sex, age, geographical location, smoking, and alcohol intake influenced serum selenium concentrations, clinically low serum selenium concentrations are not common in the USA. PMID- 12755497 TI - Induction of apoptosis by sodium selenite in human acute promyelocytic leukemia NB4 cells: involvement of oxidative stress and mitochondria. AB - The mechanisms involved in the anti-carcinogenic activity of selenium remained to be elucidated. In the present study, we examined sodium selenite induced apoptosis and oxidative stress in human acute promyelocytic leukemia cell lines (NB4). Cell growth and viability were assessed by trypan blue exclusion and cell counting; apoptosis by DNA electrophoresis and analysis of intracellular DNA contents; reactive oxygen species and reduced glutathione in the cell were measured by lucigenin dependent chemoluminescent (CL) test and spectrophotometer; mitochondrial transmembrane potential was measured by flow cytometry. Sodium selenite could inhibit the growth and induce apoptosis of NB4 cells. Sodium selenite could increase the production of reactive oxygen species (ROS) in NB4 cells and decrease the level of intracellular reduced glutathione, but caused no change in the activity of antioxidant enzymes, superoxide dismutase (SOD), glutathione peroxidase (GPx). Sodium selenite enhanced the collapse of mitochondrial transmembrane potential (MTP), in parallel with the production of ROS. Finally antioxidant N-acetylcysteine (NAC) could inhibit the ROS production, MTP collapse and apoptosis in NB4 cells. Our results suggested that sodium selenite could induce apoptosis of NB4 cells through mitochondrial change mediated by production of reactive oxygen species within the cells. PMID- 12755498 TI - Influence of low dietary calcium during pregnancy and lactation on zinc levels in maternal blood and bone in rats. AB - The effect of low dietary calcium on maternal zinc nutritional status was studied. Two groups of 6 adult female Wistar rats were fed during pregnancy and lactation with experimental diets containing either 0.2 g (LCa) or 0.6 g (NCa) of calcium/100 g. Both diets contained/100 g: 20.0 g protein (potassium caseinate), 3.5 mg Zn, 0.6 g P. A third group (n = 6) was fed a "stock diet" (SG), containing/100 g: 24.8 g protein, 1.5 g Ca, 0.6 g P, 11.6 mg Zn. Maternal blood samples were drawn from the tail before mating (To), at delivery (D) and at weaning (W); dams were sacrificed at weaning and the right femur was excised. Determinations (atomic absorption spectrometry) were: Zinc in red blood cells (RBC), Zn and Ca in ashed femur. The results (mean +/- SD) were: RBCZn (microg/mL) at To: 8.65 +/- 1.80, which did not change in the SG or in the NCa groups, but increased significantly in the LCa group (p < 0.001) (D: 18.20 +/- 4.63; W: 26.70 +/- 6.02), regarding To. Femur Zn (microg/100 mg) showed an increase (p < 0.001) in the LCa group (30.2 +/- 2.1) regarding both SG (25.3 +/- 0.7) and NCa groups (24.1 +/- 0.7). Femur Ca (mg/100 mg) decreased (p < 0.05) in the LCa group (19.2 +/- 0.9) regarding both SG (24.0 +/- 0.5) and NCa groups (21.4 +/- 0.7) and leading to a significant increase in Zn/Ca ratio (p < 0.001) in the LCa group. Therefore, dietary calcium deficiency during pregnancy and lactation would produce an increase of Zn utilization, reflected in the increase of maternal blood Zn levels and in femur Zn content. PMID- 12755499 TI - Effects of intraperitoneally injected selenium and vitamin E in rats anesthetized with halothane. AB - Halothane, commonly used for anesthetizing humans and animals, is one of the most important volatile anesthetics and may cause the formation of free radicals during its biotransformation. Free radicals may lead to degeneration of liver cells. Vitamin E and glutathione peroxidase (GSH-Px) containing selenium are two natural antioxidants, and these may protect the cellular lipid and lipoproteins against oxidative damage caused by free radicals. Therefore, the purposes of the present study were to investigate the probable protective effects of intraperitoneally administered Se and vitamin E on liver enzymes and to determine some other hematological parameters in the halothane anesthesia of rats. All rats were randomly divided into five groups. The first group was used as a control, and physiological saline (0.9%) was intraperitoneally injected into these animals as a placebo. The second group was used as an anesthesia control group and was only anesthetized with halothane for two hours. The third group received intraperitoneally administered Se (Na2SeO3, 0.3 mg/200 g body weight), the fourth group vitamin E (dl-alpha-tocopheryl acetate, 100 mg/kg body weight), and the fifth group a Se plus vitamin E combination (Na2SeO3, 0.3 mg/200 g body weight + dl-alpha-tocopheryl acetate, 100 mg/kg body weight). The activities of aspartate aminotransferase, alanine aminotransferase and alkaline phosphatase, triglycerides, erythrocyte counts, the packet-cell volume, hemoglobin concentrations and neutrophyle rates significantly increased (p < 0.05 to p < 0.01) after halothane anesthesia and returned to near control levels after Se, vitamin E and Se plus vitamin E injections. The values of cholesterol, total protein, white blood cell counts and lymphocyte rates significantly decreased (p < 0.05 to p < 0.01) in the anesthesia control group. However, the levels of albumin, total bilirubin, creatinine, the mean corpuscular volume, the mean corpuscular hemoglobin, and the mean corpuscular hemoglobin concentration were not statistically influenced. In conclusion, we have determined that halothane anesthesia affected some liver enzymes and some other biochemical and hematological parameters. Se, vitamin E and their combination may prevent the increase of liver enzymes after halothane anesthesia. Based upon these results, Se and vitamin E may play an important role in the indication of hepatic cellular injury produced by halothane. PMID- 12755500 TI - Melatonin and pinoline prevent aluminium-induced lipid peroxidation in rat synaptosomes. AB - The serum concentrations of aluminum, a metal potentially involved in the pathogenesis of Alzheimer's disease, increase with age. Also, intense and prolonged exposure to aluminum may result in dementia. Melatonin and pinoline are two well known antioxidants that efficiently reduce lipid peroxidation due to oxidative stress. Herein, we investigated the effects of melatonin and pinoline in preventing aluminum promotion of lipid peroxidation when the metal was combined with FeCl3 and ascorbic acid in rat synaptosomal membranes. Lipid peroxidation was estimated by quantifying malondialdehyde (MDA) and 4 hydroxyalkenal (4-HDA) concentrations in the membrane suspension. Under the experimental conditions used herein, the addition of aluminum (0.0001 to 1 mmol/L) enhanced MDA + 4-HDA formation in the synaptosomes. Melatonin and pinoline reduced, in a concentration-dependent manner, lipid peroxidation due to aluminum, FeCl3 and ascorbic acid in the synaptosomal membranes. These results suggest that the indoleamine melatonin and the beta-carboline pinoline may potentially act as neuroprotectant agents in the therapy of those diseases with elevated aluminum concentrations in the tissues. PMID- 12755501 TI - Comparative study of metallic biomaterials toxicity: a histochemical and immunohistochemical demonstration in mouse spleen. AB - Metallic biomaterials available for orthopaedic purposes become essential to perform important physical activities, due to their low cost and excellent mechanical properties. In addition, they are frequently used in dentistry. However, corrosion phenomena of such devices are the main problems resulting in subsequent spreading of the elements through the whole body via lymph and blood. The spleen is the most important lymphoid organ and the only one included in the blood circulation. Thus, the aim of this study was to evaluate the short-term effects on spleen tissues of heavy metals released from stainless steel and Cr-Co Mo alloys, as well as from titanium, at histochemical and immunohistochemical levels. For this purpose, metallic suspensions were obtained by electrochemical dissolution of the mentioned biomaterials: stainless steel (Fe 490 mg/L, Cr 224 mg/L, Ni 150 mg/L), Cr-Co-Mo (Cr 200 mg/L, Co 375 mg/L), and titanium (400 mg/L). Then 0.5 ml of each solution was subcutaneously administered to male Charles River mice each 72 hours during 30 days. Cryostat sections of the spleen from all groups were submitted to routine staining with haematoxylin/eosin, peroxidase detection by 3-3' diaminobenzidine (DAB), and alkaline phosphatase anti-alkaline phosphatase (APAAP) for lymphocyte detection. Several pronounced alterations were found in the spleen architecture, as manifested by irregular features within the capsule and medulla, namely depletion of T4 and B cells. Altogether these results suggest toxic alterations within the spleen induced by some of the metallic elements, indicating that the immune system may be hampered and so interfering in the body mechanisms of defence. PMID- 12755502 TI - Cadmium exposure in tobacco workers: possible renal effects. AB - Cadmium is a nephrotoxic metal widely used in industry and the main source of Cd in general population is smoking. Considering that the source of Cd in cigarettes is the tobacco leaf, the exposure to Cd was evaluated in workers employed at a tobacco leaf processing factory. Blood and urinary Cd levels were measured by flameless atomic absorption spectrometry in 87 workers and 35 controls. Urinary enzymes, total protein, albumin and uric acid were also determined to investigate the possible nephrotoxic effects of Cd. Blood Cd levels were significantly higher in workers (1.63 +/- 1.95 microg/L) than in controls (0.91 +/- 1.15 microg/L) (p = 0.044). The increase observed in urinary Cd levels of workers was non significant (0.56 +/- 0.5 microg/g creatinine in workers and 0.46 +/- 0.5 microg/g creatinine in controls). Both in workers and in controls, subjects smoking >10 cigarettes/day showed significantly increased blood Cd levels compared to non-smokers (p = 0.000 and p = 0.011, respectively). In workers, urinary alkaline phosphatase (ALP), gamma glutamyl transferase (GGT), total protein, and uric acid were observed to be significantly increased (p = 0.013, p = 0.000, p = 0.000, p = 0.025, respectively), ALP, GGT and total protein being positively correlated with Cd in urine. In conclusion, the workers in the tobacco leaf processing factory were found to be exposed to Cd compared to the general population. The increase in the urinary enzymes and proteins suggests that an exposure to Cd affects kidney functions even below the toxic limits generally accepted. PMID- 12755503 TI - Activity of pancreatic antioxidative enzymes and malondialdehyde concentrations in rats with hyperglycemia caused by fluoride intoxication. AB - The aim of this work was to examine the effect of fluoride ions on antioxidative enzyme activity in the pancreas of rats exposed during 4 months to NaF in drinking water. The study was carried out in 30 four-week-old male Wistar FL rats, that were randomly assigned to three equal groups and given distilled water ad libitum for three weeks. Subsequently, two examined groups of animals were exposed to NaF in drinking water: group 1 (10 rats) at 50 mg F(-)/L (2.63 mmol/L), group 2 (10 rats) at 100 mg F(-)/L (5.26 mmol/L). The control group (10 rats) received distilled water. After 4 months the animals were anesthetized with ether prior to collection of pancreas and cardiac blood. Serum concentrations of glucose and fluoride, as well as activities of the cytoplasmic (CuZn-SOD) and the mitochondrial (Mn-SOD) superoxide dismutase, glutathione peroxidase (GSH-Px) and concentrations of malondialdehyde (MDA) in the homogenized pancreas were measured. The activity of CuZn-SOD was reduced by 50% and a tendency to lower activities of Mn-SOD was observed. No changes were noted in the activity of GSH Px or concentrations of MDA. We conclude that: 1) the fluoride caused hyperglycemia in rats in this study is not accompanied by an activation of the free radical production in the pancreas; 2) the hyperglycemia in the exposed rats cannot be attributed to pancreatic damage caused by fluoride ions (the cause in this case appears to be extrapancreatic); 3) the inhibition of pancreatic CuZn SOD is probably due to the direct action of fluoride on the enzyme. PMID- 12755504 TI - The biceps femoris short head muscle flap: an experimental anatomical study. AB - The vascular communication between the heads of the biceps femoris muscle has been established after 25 cadaveric dissections. Perfusions of dye through the long or the short head consistently showed 1-2 anastomotic bundles. Outflow of dye opposite to the site of the perfused head was remarkable in most cases. Intramuscular dissections disclosed broad and well structured vascular networks in all short heads, but this was not true for all long heads. Our observations suggest that the anastomotic vessels alone might support the short head which, when released from its profunda femoris vessels, is adequate to cover lateral knee defects. Depending on the level of the anastomotic vessels, the proximal or the distal part of the short head should be used. A pedicled flap may be used as well, whereas transsection of the biceps tendon offers additional mobility. PMID- 12755505 TI - Orbital fractures in craniofacial trauma in Goteborg: trauma scoring, operative techniques, and outcome. AB - In the 10-year period 1986-1996, 85 patients were admitted to our unit with craniofacial injuries, 56 of whom had orbital fractures. These were studied with respect to the type of injury, type and location of fracture, presence of ocular and intracranial injury, and associated injuries to the head and body, as well as operative techniques used. Both the patient's and the surgeon's opinion on the aesthetic result were noted. The patients were also given a questionnaire about their quality of life after the injury. Road traffic crashes accounted for 31 (55%) of the injuries, falls for 9 (15%), and horse-riding for 6 (11%). The Injury Severity Score (ISS) ranged from 4 to 41 (mean 18). Twelve also had eye injuries, which resulted in complete blindness in one eye in 4 (7%). Thirty patients had 41 neurological injuries (54%), frontal contusions being the most frequent diagnosis (n = 15). Exact repositioning with rigid fixation included bone grafting to the orbit in 11 patients, and the dominating bone graft was split calvarium (n = 5). Forty-two patients completed a questionnaire, 26 of whom (64%) had no aesthetic complaints. Seven of the 42 were too disabled to work one year after the injury. Re-exploration was infrequent and the aesthetic outcome, both in the surgeon's and the patient's opinion, was good. However, the older the patient, the worse the outcome. PMID- 12755506 TI - Need for facial reanimation after operations for vestibular schwannoma: patients perspective. AB - A total of 779 patients operated on for vestibular schwannoma mostly by the translabyrinthine approach in Denmark during the period 1976-2000 answered a questionnaire about various postoperative consequences. In this paper we describe the patients' facial function evaluated by professionals one year postoperatively and self-evaluated by each patient according to the House-Brackmann scale at the time of the questionnaire. The patients' self-evaluation was more pessimistic than that of the professionals with 26% reporting House-Brackmann grade IV-VI, compared with 20%. One hundred and seventeen (15%) of 779 patients considered their facial palsy to be a big problem and 125 patients (16%) were interested in surgical treatment for the sequelae of facial palsy. Seventy-eight (10%) had already had some kind of operation, usually the VII-XII coaptation. Thirty-three of 61 patients who had already been operated on for facial palsy were interested in further surgical treatment. One hundred and ninety-five patients (25%) had some kind of operation on the eye, mostly (88%) a tarsorrhaphy. Reanimation procedures such as a palpebral gold weight or a spring, apparently still have a small place in Denmark. In conclusion, there seem to be a considerable and unmet need for surgical reanimation of facial function in patients with facial palsy after operations for vestibular schwannoma in Denmark. PMID- 12755507 TI - Restoration of involuntary tonic contraction of the levator muscle in patients with aponeurotic blepharoptosis or Horner syndrome by aponeurotic advancement using the orbital septum. AB - Muller's muscle can be thought of as a large serial type of muscle spindle of the levator muscle. Effective stretching of the mechanoreceptor in the proximal part of Muller's muscle by voluntary phasic contraction of the levator muscle for initial opening of the eye induces involuntary tonic contraction of the levator muscle as a stretch reflex via the mesencephalic trigeminal nucleus, to maintain an adequate visual field. After disinsertion of the levator aponeurosis from the tarsus by habitual rubbing, elongation of Muller's muscle secondary to thinning (aponeurotic blepharoptosis) or paralysis (Horner syndrome) desensitises the mechanoreceptor of Muller's muscle, resulting in blepharoptosis. Shortening of the elongated and thinned Muller's muscle by instillation of phenylephrine, and surgical shortening, and fixation of the disinserted, elongated, and thinned aponeurosis using the orbital septum, restored involuntary tonic contraction of the levator muscle in nearly all of 2000 patients with aponeurotic blepharoptosis and in 11 patients with Horner syndrome. PMID- 12755508 TI - Combined surgical lipectomy and liposuction in the treatment of benign symmetrical lipomatosis of the head and neck. AB - Benign symmetrical lipomatosis (Madelung syndrome) is a rare disease of unknown aetiology, which is characterised by diffuse growth of non-encapsulated lipomas. Between 1995 and 2000 we treated 11 patients with benign symmetrical lipomatosis in the head and neck. The group comprised 10 men and one woman aged 34 to 62 years (mean 47). The most common complaints were reduced range of movement of the head and obstruction when eating or speaking. Combined lipectomy and liposuction were done for all patients, with liposuction being done at a second session. The mean follow-up period was 2.7 years. The functional results were satisfactory in all patients. Nine of the 11 patients were also satisfied with the aesthetic outcome. Two patients developed recurrence 1.5 and 2 years after the operation, respectively. There were no serious complications. We think that combined lipectomy and liposuction is a successful procedure for treating benign symmetrical lipomatosis in the head and neck region. Nevertheless, the advantages and drawbacks of the two techniques should be considered preoperatively. PMID- 12755509 TI - Vaginal construction with skin grafts and vacuum-assisted closure. AB - Probably the most common method of constructing a vagina in patients with the Mayer-Rokitansky-Kuster syndrome is the technique popularised by McIndoe and Banister in 1938. A cavity is created between the rectum and urethra-bladder complex and is lined with split-thickness skin grafts. One of the disadvantages of using split-thickness skin grafts is the incidence of late contraction of the neovagina. To avoid this problem full-thickness skin grafts have been used, but their take is less reliable. A new technique to improve the take of skin grafts is the VAC-system (vacuum assisted closure, KCI) which has proved to be particularly valuable in grafting difficult anatomical sites. We have used the VAC-system in the construction of a vagina in one case with split-thickness skin grafts and in two cases with full-thickness skin grafts. In all three cases the take was excellent with little discomfort for the patients. It was not necessary to stent the neovagina in the postoperative period and coitus was possible within a month of operation. PMID- 12755510 TI - Repair of giant incisional hernias with polypropylene mesh: a retrospective study. AB - The aim of the present study was to evaluate the long term results of repair of ventral incisional hernias or of defects in the abdominal wall using polypropylene mesh. Eighty-eight patients were operated on from 1979-1996, inclusive. Abdominal protrusion was found in 78 patients. Fifty-one of these patients had previously had an incisional hernia repaired and the remaining 10 patients had an abdominal wall defect as a result of excision of a malignant tumour. The polypropylene mesh was placed extraperitoneally and sutured with two rows of interrupted stitches, using non-absorbable sutures. Recurrence of the hernia was found in 10 of the 67 patients with incisional hernia. Eight patients had a relaxation of the muscles of the abdominal wall. Perioperative complications consisted of infection (n = 4), embolism (n = 1), haematoma (n = 1), and pneumonia (n = 3, one fatal), and one fatal bowel perforation. Median follow up time was 5.7 years (range 0-17). It is therefore possible to obtain acceptable results after repair of larger incisional hernias even if they had been repaired before. PMID- 12755511 TI - Long-term results with the proximally-based neurovascular lateral calcaneal flap. AB - From 1995 to 1997, the proximally-based neurovascular lateral calcaneal flap was used in eight patients to cover defects in the Achilles tendon area or the lateral malleolar region. The mean postoperative follow-up was 40 months (range 29-51). The outcome of the operation was investigated both clinically and using a questionnaire sent to patients. All flaps healed uneventfully within a mean of 33 days. Revisional operations were required in two cases because of pain at the donor site. No flaps had broken down up to the follow-up, which was between three and five years. Two of the patients operated on were paraplegic, and the other six patients had sensate flaps. Five of the six patients with sensate flaps were able to wear normal shoes and one patient needed a shoe elevated by about 7.5 cm because of a disease independent of the flap. All but two patients were satisfied with the functional and aesthetic results. The proximally-based neurovascular lateral calcaneal flap proved to be safe and can be recommended as a good option to cover tissue defects in the Achilles tendon area or the lateral malleolar region. PMID- 12755512 TI - Value of dynamic splinting after replacement of the metacarpophalangeal joint in patients with rheumatoid arthritis. AB - In a retrospective study, we compared a group of 13 patients (41 joints) treated with dynamic splints after replacement of the metacarpophalangeal (MCP) joints with a group of 9 patients (29 joints) not so treated. We failed to confirm our hypothesis, that the range of movement in the joints would be less in the group treated with dynamic splints. Furthermore, residual extension lag was significantly less (p = 0.002) in the treated group. We conclude that postoperative dynamic splinting seems to be useful after replacement of MCP joints with silastic implants. PMID- 12755513 TI - Primary repair of the collateral ligament of the proximal interphalangeal joint using a suture anchor. AB - Ten out of 12 complete ruptures of the collateral ligament of the proximal interphalangeal joint were repaired primarily using a suture anchor. Active finger exercise was started gently by taping the injured finger to adjacent one one week postoperatively. All patients returned to their original occupation or sports activities within seven weeks of operation. No patients complained of pain on the involved finger at rest or during movement. There was no instability of the proximal interphalangeal joint on manual lateral stress test postoperatively. PMID- 12755514 TI - Subungual glomus tumours of the hand: diagnosis and treatment of 14 cases. AB - A glomus tumour is a rare tumour of the hand, which presents with a classic triad of symptoms of temperature sensitivity, pain, and localised tenderness. We report a series of 14 patients with glomus tumours in the subungual region. There were four men and 10 women, whose mean age was 46 years (range 28-65). All patients complained of pain and localised tenderness and six of temperature sensitivity. The bony lesion was seen on radiographs in four patients. We used a transungual approach in all cases. A detailed history and clinical findings are the best way of making the diagnosis. The treatment of glomus tumour consists of complete excision and repair of the nailbed. Inadequate excision of the tumour usually causes local recurrence. PMID- 12755515 TI - Massive vascular leiomyoma of the hand. AB - A patient presented with a massive (5.0 x 4.0 x 3.0 cm) vascular leiomyoma (angioleiomyoma) in the hand, which had developed over a 50-year-period without pain. The clinical presentation, microscopic findings, and behaviour of vascular leiomyoma are described. PMID- 12755516 TI - The structure of science information. AB - The organization of information within science can be investigated in a principled way through analysis of science language. The restricted use of language in science enables description of the informational structure of science and of particular subfields, with strong similarities to structures in mathematics and programming languages. This result rests on decades of research into the relation between form and content in language, based on an information theoretic approach to the structure of information. Examples are provided from immunology and the social sciences. Practical applications include storage of science information in databases, indexing the literature, and identification and resolution of controversy. PMID- 12755517 TI - Two biomedical sublanguages: a description based on the theories of Zellig Harris. AB - Natural language processing (NLP) systems have been developed to provide access to the tremendous body of data and knowledge that is available in the biomedical domain in the form of natural language text. These NLP systems are valuable because they can encode and amass the information in the text so that it can be used by other automated processes to improve patient care and our understanding of disease processes and treatments. Zellig Harris proposed a theory of sublanguage that laid the foundation for natural language processing in specialized domains. He hypothesized that the informational content and structure form a specialized language that can be delineated in the form of a sublanguage grammar. The grammar can then be used by a language processor to capture and encode the salient information and relations in text. In this paper, we briefly summarize his language and sublanguage theories. In addition, we summarize our prior research, which is associated with the sublanguage grammars we developed for two different biomedical domains. These grammars illustrate how Harris' theories provide a basis for the development of language processing systems in the biomedical domain. The two domains and their associated sublanguages discussed are: the clinical domain, where the text consists of patient reports, and the biomolecular domain, where the text consists of complete journal articles. PMID- 12755518 TI - Information extraction for enhanced access to disease outbreak reports. AB - Document search is generally based on individual terms in the document. However, for collections within limited domains it is possible to provide more powerful access tools. This paper describes a system designed for collections of reports of infectious disease outbreaks. The system, Proteus-BIO, automatically creates a table of outbreaks, with each table entry linked to the document describing that outbreak; this makes it possible to use database operations such as selection and sorting to find relevant documents. Proteus-BIO consists of a Web crawler which gathers relevant documents; an information extraction engine which converts the individual outbreak events to a tabular database; and a database browser which provides access to the events and, through them, to the documents. The information extraction engine uses sets of patterns and word classes to extract the information about each event. Preparing these patterns and word classes has been a time-consuming manual operation in the past, but automated discovery tools now make this task significantly easier. A small study comparing the effectiveness of the tabular index with conventional Web search tools demonstrated that users can find substantially more documents in a given time period with Proteus-BIO. PMID- 12755519 TI - Rutabaga by any other name: extracting biological names. AB - As the pace of biological research accelerates, biologists are becoming increasingly reliant on computers to manage the information explosion. Biologists communicate their research findings by relying on precise biological terms; these terms then provide indices into the literature and across the growing number of biological databases. This article examines emerging techniques to access biological resources through extraction of entity names and relations among them. Information extraction has been an active area of research in natural language processing and there are promising results for information extraction applied to news stories, e.g., balanced precision and recall in the 93-95% range for identifying person, organization and location names. But these results do not seem to transfer directly to biological names, where results remain in the 75-80% range. Multiple factors may be involved, including absence of shared training and test sets for rigorous measures of progress, lack of annotated training data specific to biological tasks, pervasive ambiguity of terms, frequent introduction of new terms, and a mismatch between evaluation tasks as defined for news and real biological problems. We present evidence from a simple lexical matching exercise that illustrates some specific problems encountered when identifying biological names. We conclude by outlining a research agenda to raise performance of named entity tagging to a level where it can be used to perform tasks of biological importance. PMID- 12755520 TI - Information extraction from biomedical text. AB - Information extraction is the process of scanning text for information relevant to some interest, including extracting entities, relations, and events. It requires deeper analysis than key word searches, but its aims fall short of the very hard and long-term problem of full text understanding. Information extraction represents a midpoint on this spectrum, where the aim is to capture structured information without sacrificing feasibility. One of the key ideas in this technology is to separate processing into several stages, in cascaded finite state transducers. The earlier stages recognize smaller linguistic objects and work in a largely domain-independent fashion. The later stages take these linguistic objects as input and find domain-dependent patterns among them. There are now initial efforts to apply this technology to biomedical text. In other domains, the technology plateaued at about 60% recall and precision. Even if applications to biomedical text do no better than this, they could still prove to be of immense help to curatorial activities. PMID- 12755521 TI - Paraphrasing for condensation in journal abstracting. AB - When authors of empirical science articles write abstracts, they employ a wide variety of distinct linguistic operations which interact to condense and rephrase a subset of sentences from the source text. An on-going comparison of biological and biomedical journal articles with their author-written abstracts is providing a basis for a more linguistically detailed model of abstract derivation using syntactic representations of selected source sentences. The description makes use of rich dictionary information to formulate paraphrasing rules of differing degrees of generality, including some which are sublanguage-specific, and others which appear valid in several languages when formulated using "lexical functions" to express important semantic relationships between lexical items. Some paraphrase operations may use both lexical functions and rhetorical relations between sentences to reformulate larger chunks of text in a concise abstract sentence. The descriptive framework is computable and utilizes existing linguistic resources. PMID- 12755522 TI - Determination of sensitivity and specificity of breast tumor diagnosis by primary health care providers (Behvarz) using clinical examination by obstetrician as a gold standard. AB - AIM: The aim of this study was to find a practical screening strategy to detect breast tumors in those who cannot refer to specialists due to problems, such as geographic location, and economical obstacles. METHODS: Considering the mentioned aim this study was designed to compare the sensitivity and specificity of diagnosis of breast tumors made by health care providers (Behvarz) with those made by specialists. For this, the results of examinations of Behvarzes and an obstetrician on 2000 women referring to the 17 health care centers of Kerman and Zarand cities were compared. RESULTS: Among the 2000 women examined by Behvarzes 170 cases were reported to have pathological signs (palpable mass) and 1830 cases were reported without any pathological sign. Among 169 cases diagnosed by physicians as having pathological signs, 162 cases had also been diagnosed by Behvarzes, and there were only seven cases diagnosed by physicians that had been missed by Behvarzes. There were eight cases diagnosed by Behvarzes as having pathological signs that were reported healthy by physicians. CONCLUSIONS: Based on these findings, the sensitivity and specificity of diagnosis of breast tumors made by Behvarzes was 95.8% and 99.56%, respectively, compared with those made by specialists. Considering the obtained results, the screening program for breast tumors by Behvarzes can be very helpful in early diagnosis of breast tumors. PMID- 12755523 TI - Bilateral seminomas in a 45X/46XY mosaic with Turner's phenotype: an unusual case of mixed gonadal dysgenesis. AB - A wide spectrum of phenotypic manifestations are seen in cases with 45X/46XY mosaicism. We present a case with 45X/46XY having female phenotype with Turner's stigmata. Prophylactic laparoscopic gonadectomy was performed and the patient was found to have mixed gonadal dysgenesis with bilateral gonadoblastomas. Microinvasive seminomas were also detected in both gonadoblastomas. The presence of Y cell line in karyotype prompted early and prophylactic gonadectomy, a procedure which is life-saving for these individuals. PMID- 12755524 TI - Pregnancy associated aplastic anemia: maternal and fetal outcome. AB - AIM: To study the maternal and fetal outcome in cases of aplastic anemia associated with pregnancy. METHOD: Retrospective analysis of seven cases of aplastic anemia diagnosed during pregnancy. RESULT: Four patients had an overall successful pregnancy outcome. These were all cases of non-severe aplastic anemia. There was one case each of intrauterine and neonatal death in the severe aplastic anemia group. There was one maternal mortality. CONCLUSION: Maternal and fetal outcome is poor in severe aplastic anemia. PMID- 12755525 TI - Effect of dietary habits on prevalence of anemia in pregnant women of Delhi. AB - AIM: To see the effect of various dietary habits, such as a vegetarian diet or various types of meat, on the prevalence of anemia in pregnant women. METHODS: A study was carried out in Delhi to determine the effect of different dietary habits on prevalence of anemia during pregnancy by questioning the women during pregnancy regarding their dietary habits (vegetarian diet, jhatka or halal meat) and assessing their hemoglobin levels. The data was compiled and chi2 test was employed for understanding the associations between the effect of food habits on prevalence of anemia. RESULTS: Mean age was 26.5 years. Most women were in the second (26%) or third trimester (63.2%) of pregnancy. Prevalence of anemia was found to be very high. Of 1150 women, 96% were anemic (89.8% mildly anemic, 5.3% severely anemic). Anemia was seen in 96.18% cases in vegetarian women, 95.3% in halal meat eaters, and 96.2% in jhatka meat eaters (not significant). Although the percentage of women with < 11 g/dL Hb was less in the jhatka group eating meat more than 5 times per month, than in halal meat eaters and vegetarians, the difference was not statistically significant. CONCLUSIONS: There is very high prevalence of anemia during pregnancy in Delhi, probably due to very low frequency of meat eating in India. Different types of dietary habits had no effect on the prevalence of anemia in pregnant Indian women. PMID- 12755526 TI - A randomized controlled study of three targets of propofol plasma concentration in patients undergoing uterine dilation and curettage. AB - AIM: To compare the efficacy and the complications of three target levels of propofol plasma concentration in patients undergoing uterine dilation and curettage. METHODS: Sixty-nine patients were randomly allocated to receive propofol target controlled infusion at different target concentrations of 4 (group I), 5 (group II) and 6 (group III) mcg/mL combined with 1 microg/kg of fentanyl and 66% of nitrous oxide. Patients' movement during the procedure, hemodynamic variables, oxygen saturation, end tidal carbon dioxide, time to sleep and awake, and bispectral index score were recorded. RESULTS: Seven patients in group I, one in group II, and none in group III (P < 0.05) moved grossly during the procedure. More patients in group III developed hypotension (5 vs 0 and 2 in group I and II, P < 0.05), but no difference was found regarding respiratory complication. No difference was found in time to sleep, but time to wake was longer in group III. CONCLUSION: Propofol infusion at the target concentration of 5 microg/mL was recommended for uterine dilation and curettage when it was administered with fentanyl 1 microg/kg and nitrous oxide 66% under close monitoring and appropriate respiratory management. PMID- 12755527 TI - Immunogenicity and safety of two schedules of Hepatitis B vaccination during pregnancy. AB - AIM: To test the immunogenicity and safety of HepatitisB (HB) vaccination during pregnancy and passive transfer of antibodies to the newborn. METHODS: Ninety-nine HBsAg-negative pregnant women were divided randomly in two groups; group I was given two, and group II was given three doses of recombinant Hepatitis B vaccine and followed up until 4 months after delivery. RESULTS: The vaccination was safe. The maternal anti-HB antibody levels at delivery were significantly higher in three dose schedule as compared to two dose schedule. Similarly, the antibody levels in the newborns were higher in the three dose schedule. The maternal antibody levels were higher at 2 and 4 months post-delivery in group II but were observed to be declining in the newborns. CONCLUSION: Hepatitis B vaccination during pregnancy is safe and highly immunogenic, and there is passive transfer of antibodies to the newborns. PMID- 12755528 TI - A comparison between intravaginal and oral misoprostol for labor induction: a randomized controlled trial. AB - AIM: To compare the effectiveness and safety between intravaginal and oral misoprostol for labor induction. METHODS: One hundred and six pregnant women at term with unfavorable cervix (Bishop score < or = 4) and no contraindication to prostaglandin therapy were randomized to receive either intravaginal misoprostol 50 microg every 4 h or oral misoprostol 50 microg every 4 h for prospective randomized controlled trial study. Treatment interval from induction to vaginal delivery, maternal and neonatal complications were the main outcome measures. RESULTS: There were no statistical differences of baseline characteristics and Bishop score prior to intervention between both groups. Time interval from induction to vaginal delivery in the oral group was slightly, but significantly, longer than that of the intravaginal group (886.1 +/- 443.5 min vs 637.0 +/- 373.3 min, respectively.) Additionally, the number of doses required was significantly higher in the oral group. Nonetheless, there was no significant difference between both groups with regard to failure of induction and maternal neonatal complications. CONCLUSION: The effectiveness in terms of failed induction and safety were comparable between intravaginal and oral misoprostol, but intravaginal route was better with respect to treatment interval and number of required doses. Both routes of administration can alternatively be used for labor induction. PMID- 12755529 TI - Woman with postpartum ventricular tachycardia and hypomagnesemia. AB - A 28-year-old Japanese woman who had received continuous intravenous infusion of magnesium sulfate from 24 weeks of pregnancy until delivery underwent cesarean section at 30 weeks and gave birth to twins. Serum magnesium sharply declined to a subnormal level of 1.5 mg/dL on postpartum day 4. The patient exhibited sinus bradycardia (48 b.p.m.) with intermittent supraventricular contraction on postpartum day 2, intermittent ventricular bigeminy on postpartum day 3, and frequent selfterminated polymorphic ventricular tachycardia on postpartum day 4. The electrocardiogram (ECG) disclosed that the patient had prolonged QTc of 0.45 0.67. Correction of serum magnesium improved ECG findings promptly, resulting in the disappearance of arrhythmias. Hypomagnesemia due to postpartum diuresis may have played a role causing ventricular tachyarrhythmia in this patient. PMID- 12755530 TI - Nutritional status of pregnant and lactating women in Japan: a comparison with non-pregnant/non-lactating controls in the National Nutrition Survey. AB - AIM: To describe the nutritional status in Japanese pregnant and lactating women at a national level, through a comparison with their non-pregnant/non-lactating controls. MATERIAL AND METHODS: Pooled data from five years (1995-1999) of the National Nutrition Survey, Japan. Data on 330 pregnant and 388 lactating women, and their one-by-one matched non-pregnant/non-lactating controls, were used for analysis. RESULTS: There were fewer smokers, drinkers, and exercisers in pregnant women compared to their controls (P < 0.01). Both pregnant and lactating women showed significantly higher intakes of carbohydrates, calcium and vitamin B2. Mean iron intakes ranged 10.3-11.5 mg in the four groups, all being lower than the recommended intake level for non-pregnant/non-lactating women (12mg/day). Pregnant women consumed more fruits, milk and milk products, and less alcohol beverages and fish/shellfish compared to controls. Lactating women consumed more grain, vegetables, milk and milk products, and less alcohol beverages. There were 22.9% anemic subjects (Hb < 11 g/dL) in pregnant women, and 11.1% anemic subjects (Hb < 12g/dL) in lactating women and 15.7% in non-pregnant/non-lactating women. None of the pregnant subjects was severely anemic (Hb < 8 g/dL). No significant differences were observed in iron intakes between anemic and non-anemic women in each group. CONCLUSION: Pregnant and lactating women in Japan were aware of adopting healthy behaviors, such as smoking less, drinking less, and taking more milk or milk products compared to controls. Prevalence of mild anemia in pregnant women may have been partly due to plasma volume expansion in pregnancy. PMID- 12755531 TI - Use of fetal-pelvic index in the prediction of vaginal birth following previous cesarean section. AB - AIM: To clarify the usefulness of the fetal-pelvic index as a predictor of vaginal birth after previous lower segment cesarean section. METHODS: One hundred and seventy women with one lower segment cesarean section who attempted for trial of vaginal birth were enrolled. Pelvimetry was performed to measure maternal pelvic inlet and mid-cavity circumferences at 37 weeks gestation. Ultrasound was performed to measure fetal head and abdominal circumferences at 38-39 weeks. The fetal-pelvic index was derived. The predictability of fetal-pelvic index in the predicting the outcome of delivery was calculated. RESULTS: Fifty-seven (33.5%) women required repeated cesarean section and 113 (66.5%) delivered vaginally. Twenty-two women with positive fetal-pelvic index had repeated cesarean section. The predictability of positive fetal-pelvic index was 48.9%. Ninety of the 125 patients with a negative fetal-pelvic index delivered vaginally. The predictability of negative fetal-pelvic index was 72.0%. CONCLUSIONS: Fetal pelvic index derived in the antenatal period has low predictive value in predicting of successful vaginal birth after cesarean section. This index is not useful in clinical practice. PMID- 12755532 TI - Effect of hypoxia on the auditory system of goat fetuses during extrauterine incubation. AB - AIM: To investigate the effect of hypoxia on the auditory system in fetuses, we attempted to analyze the auditory brainstem response, the middle latency response, and changes of several physiological parameters of goat fetuses during extrauterine incubation. METHODS: We conducted extrauterine incubation of five goat fetuses at around 127days of gestation (term = 148 days). Their physiological parameters, such as fetal heart rate, mean blood pressure, flow rate of carotid artery, as well as the auditory brainstem response and middle latency response, were recorded prior to and during hypoxia, and the two sets of data were compared with each other. RESULTS: In all five cases, the fetal heart rate decreased from 178 +/- 12.2 b.p.m. to 144 +/- 15.2 b.p.m. during hypoxia, while mean blood pressure and flow rate of carotid artery increased from 37.3 +/- 3.7 mmHg to 43.2 +/- 5.1 mmHg, and from 38.5 +/- 5.5mL/min to 47.0 +/- 5.1 mL/min, respectively. The latency of the auditory brainstem response's wave V and of the middle latency response's Pa wave elongated from 5.24 +/- 0.24 ms to 5.69 +/- 0.20 ms, and from 19.2 +/- 1.6 ms to 20.9 +/- 1.4 ms, respectively. CONCLUSIONS: Although fetal compensatory reactions, such as increases in mean blood pressure and flow rate of carotid artery during hypoxia were recognized, elongation of latency, and decrement of amplitude were observed in the auditory brainstem response and middle latency response. These results suggest that hypoxia itself influences the auditory system of the fetus. PMID- 12755533 TI - On practicing the art of clinical research with medical students. PMID- 12755534 TI - Long-residence-time nano-scale liposomal iohexol for X-ray-based blood pool imaging. AB - RATIONALE AND OBJECTIVES: Although soluble nonionic iodine compounds with low systemic toxic effects have been developed for use in computed tomography (CT), they have short residence times of a few minutes or mere seconds-insufficient time for blood pool imaging, even with high-speed multi-detector row spiral CT. Moreover, potential renal toxic effects preclude repeated administration of these contrast agents during imaging, as well as their use in patients with compromised renal function. The objective of this study was to develop and evaluate a CT contrast agent for blood pool imaging that remains in the blood for more than 3 hours and that is relatively nontoxic to the kidneys. MATERIALS AND METHODS: The authors assessed a liposomal iohexol formulation for its encapsulation efficiency in terms of milligrams of iodine per milliliter of lipid formulation and for its stability in phosphate buffer solution and in human plasma in vitro. Using a rabbit model, they also assessed the formulation's in vivo stability, residence time, and enhancement of contrast on images of various organ systems. RESULTS: The formulation, which contained 34.8 mg of iodine per milliliter of liposomal iohexol solution, remained stable in blood plasma both in vitro and in vivo, after injection into rabbit vasculature. An intravenous dose of 475 mg of iodine per kilogram of body weight produced contrast enhancement in the rabbit model of approximately 130 HU in the aorta and liver cortex and approximately 100 HU in the kidney cortex. Contrast enhancement was maintained for 3 hours after injection, and minimal clearance of the contrast agent via the kidneys was observed. CONCLUSION: The liposomal iohexol formulation tested in this study had a sufficient residence time for blood pool imaging in a rabbit model. Future experiments with long-residence-time iohexol formulations may lead eventually to applications in cardiac imaging and in early tumor detection. PMID- 12755535 TI - Detection and quantification of breast tumor necrosis with MR imaging: value of the necrosis-avid contrast agent Gadophrin-3. AB - RATIONALE AND OBJECTIVES: The authors evaluated the use of T1-weighted magnetic resonance (MR) imaging with Gadophrin-3 enhancement and of plain T2-weighted MR imaging to detect and quantify breast tumor necrosis. MATERIALS AND METHODS: Twenty EMT-6 tumors (mouse mammary sarcoma), implanted into the mammary fat pad of BALB/c-AnNCrl mice, underwent MR imaging with plain T2-weighted and T1 weighted fast field echo sequences before and 24 hours after injection of Gadophrin-3, a new necrosis-avid contrast agent. Tumor necrosis on MR images was quantified by means of a dedicated segmentation program and was correlated with histologic findings. RESULTS: In all tumors a central necrosis was revealed by histopathologic analysis, and central enhancement was seen with Gadophrin-3 on T1 weighted images. Small tumors (diameter, < 1 cm) showed an inhomogeneous central enhancement, whereas larger tumors (diameter, > 1 cm) enhanced mainly in the periphery of necrotic tissue. Plain T2-weighted images showed a hyperintense central area in only three of 20 cases with a large central necrosis. CONCLUSION: Gadophrin-3-enhanced T1-weighted images are superior to plain T2-weighted images for the detection of necrosis in a murine tumor xenograft model. PMID- 12755536 TI - Evaluation of simethicone-coated cellulose as a negative oral contrast agent for abdominal CT. AB - RATIONALE AND OBJECTIVES: Because of the increased clinical use of computed tomography (CT) for imaging the abdominal vasculature and urinary tract, there is a need for negative contrast agents. The authors undertook this study to assess the suitability of simethicone-coated cellulose (SCC), which is approved for use as an oral contrast agent in sonography, for use as a negative oral contrast agent in abdominal CT. MATERIALS AND METHODS: This prospective study involved 40 adult patients scheduled to undergo abdominal CT for the evaluation of hematuria. Prior to scanning, 20 subjects received 800 mL of SCC and 20 received 800 mL of water as an oral contrast agent. Imaging was performed with a multi-detector row helical scanner in two phases, according to the abdominal CT protocol used for hematuria evaluation at the authors' institution. The first, "early" phase began an average of 15 minutes after the ingestion of contrast material; the second, "late" phase began an average of 45 minutes after the ingestion of contrast material. Blinded analysis was performed by three abdominal radiologists separately, using a three-point scale (0 = poor, 1 = acceptable, 2 = excellent) to assess the effectiveness of SCC for marking the proximal, middle, and distal small bowel. Average scores for enhancement with SCC and with water were obtained and compared. Statistical analysis was performed with a Wilcoxon signed-rank test. RESULTS: SCC was assigned higher mean scores than water for enhancement in each segment of the bowel, both on early-phase images (0.8-1.35 for SCC vs 0.6 1.1 for water) and on late-phase images (1.1-1.4 vs 0.81-0.96). Bowel marking with SCC, particularly in the jejunum and ileum, also was rated better than that with water in a high percentage of patients. The differences between the scores for water and for SCC, however, were not statistically significant (P > .05). CONCLUSION: SCC is effective as a negative oral contrast agent for small bowel marking at CT. PMID- 12755537 TI - Delayed 99mTc-labeled erythrocyte scintigraphy in patients with lower gastrointestinal tract hemorrhage: effect of positive findings on clinical management. AB - RATIONALE AND OBJECTIVES: This study was performed to determine whether the results of delayed technetium 99m (99mTc)-labeled erythrocyte scintigraphy for lower gastrointestinal tract hemorrhage resulted in different clinical management and outcome from that in cases in which the results of initial scintigraphy were negative or equivocal. MATERIALS AND METHODS: The authors retrospectively reviewed all 398 99mTc-labeled erythrocyte scintigraphic studies obtained emergently for lower gastrointestinal tract hemorrhage at their institution between January 1, 1994, and December 7, 2001. Of 67 patients who underwent delayed studies, 37 had positive findings (average delay, 18.4 hours; range, 6-25 hours) and 30 had negative findings (average delay, 20.1 hours; range, 8-26 hours). Clinical management and outcome were compared between these two groups with respect to duration of hospitalization, volume of blood transfusion, mortality, and the percentage who were treated conservatively or referred for angiography, endoscopy, and/or surgery. RESULTS: Patients with positive delayed studies were referred more frequently for angiography than those with negative studies (35% vs 0%, P < .01). There were no significant differences between patients with positive findings and patients with negative findings with respect to mortality (8% vs 0%, P < .32), transfusion requirements (5.6 vs 3.2 units, P < .20), hospitalization (9.5 vs 6.1 days, P < .11), the percentage treated conservatively (35% vs 37%, P < .90), or the percentages referred for endoscopy (49% vs 60%, P < .50) or for surgery (24% vs 17%, P < .64). CONCLUSION: Positive findings at delayed scintigraphy resulted in increased referrals for angiography but had no other effect on clinical course or outcome of lower gastrointestinal tract hemorrhage. PMID- 12755538 TI - Operating characteristics of hyperpolarized 3He and arterial spin tagging in MR imaging of ventilation and perfusion in healthy subjects. AB - RATIONALE AND OBJECTIVES: The authors tested the feasibility of a magnetic resonance (MR) imaging method combining the use of hyperpolarized helium 3 (3He) for ventilation imaging and an arterial spin-tagging sequence for perfusion imaging in six healthy human subjects. MATERIALS AND METHODS: High-resolution sagittal images depicting 3He distribution were acquired after the subjects' inhalation of 500 mL of laser-hyperpolarized 3He produced by spin-exchange optical pumping. Perfusion MR imaging was performed with a steady-state arterial spin-tagging sequence that enabled the acquisition of three-dimensional images of pulmonary perfusion without the need for subject breath holding. RESULTS: The 3He ventilation images display, with high signal intensity and detailed anatomic localization, the airspace of the lung parenchyma. The signal intensity on the perfusion images decreased by 23.2% with the use of arterial spin tagging. Ventilation and perfusion were matched, as is expected in healthy subjects. CONCLUSION: This method may have important applications in the assessment of lung function, enabling the calculation of regional ventilation-perfusion ratios. It may also aid in the selection of candidates for lung volume-reduction surgery. PMID- 12755539 TI - MR imaging for the detection of endoleaks in recipients of abdominal aortic stent grafts with low magnetic susceptibility. AB - RATIONALE AND OBJECTIVES: This study was performed to assess the efficacy of magnetic resonance (MR) imaging for the detection of endoleaks in recipients of abdominal aortic stent-grafts with low magnetic susceptibility. MATERIALS AND METHODS: A retrospective search was conducted in radiology department records for cases of patients with low-susceptibility stent-grafts who had been evaluated with MR imaging and either computed tomography (CT) or conventional angiography within a 1-month time frame. Any endoleaks previously confirmed and classified with the use of CT and/or conventional angiography were compared with findings from MR imaging. RESULTS: Nine patients fit the selection criteria. Images of five of those patients depicted six different endoleaks. Two endoleaks had been confirmed with CT, another two had been confirmed with CT and angiography, and two had been confirmed with angiography alone. All endoleaks visualized at CT and/or angiography were accurately detected and classified also with MR imaging. In some cases, the endoleak was more clearly visualized with MR imaging than with CT. In four patients in whom no endoleaks were found at CT, MR imaging also indicated no endoleaks. CONCLUSION: MR imaging is a suitable modality for identifying endoleaks in patients with low-susceptibility stent-grafts. Moreover, MR imaging may be more sensitive than CT for the detection of small endoleaks. PMID- 12755540 TI - A randomized trial of prophylactic acetylcysteine and theophylline compared with placebo for the prevention of renal tubular vacuolization in rats after iohexol administration. AB - RATIONALE AND OBJECTIVES: Renal tubular vacuolization (RTV), which has been shown to occur after the use of iodinated contrast material, may be one of the earliest signs of contrast medium-induced renal injury. In this study, the authors tested a method for preventing RTV with the administration of acetylcysteine, theophylline, or both, prior to contrast medium administration. MATERIALS AND METHODS: Eighty rats were randomly selected for inclusion in the study. The treatment group consisted of three subgroups, each of which received prophylactic acetylcysteine, theophylline, or both before injection of iohexol. The control group comprised five subgroups, each of which received acetylcysteine, theophylline, both, normal saline injection, or orally administered normal saline prior to iohexol injection. RESULTS: The occurrence of RTV in the treatment groups was compared with that in the control subgroup that received normal saline and iohexol. All of the rats in that control subgroup (n = 10) and 97% of the rats in the treatment group (n = 30) developed RTV. CONCLUSION: The administration of acetylcysteine, theophylline, or both prior to iohexol injection did not prevent RTV from occurring in rats. PMID- 12755542 TI - The art of clinical research with medical students. PMID- 12755541 TI - MR angiography of the carotid arteries: parameters affecting image quality. AB - RATIONALE AND OBJECTIVES: This study was performed to evaluate the relationship between dose levels of contrast medium and image quality in magnetic resonance (MR) angiography of the carotid arteries with fluoroscopically monitored, manually triggered, elliptically ordered image acquisitions. MATERIALS AND METHODS: Twenty-five patients with clinical indications for angiography of the carotid arteries were examined with MR at 1.5 T by using a fluoroscopically monitored, manually triggered, elliptically ordered pulse sequence with the administration of one of three different volumes of gadolinium-based contrast medium. The signal intensities of the vessel lumen and the surrounding tissues were measured in single partitions at the origin of the common carotid artery, the carotid bifurcation, and the intracranial internal carotid arteries. The contrast-to-noise ratio in these regions of interest also was measured. Maximum intensity projection image quality was appraised for blurring, artifacts, venous enhancement, background suppression, and contrast medium distribution. RESULTS: No artifacts or venous enhancement was observed. The position of the fluoroscopic section affected the distribution of contrast medium along the vessel, as evidenced by the difference between the contrast-to-noise ratio at the origin of the common carotid artery and the ratio at the carotid bifurcation and the intracranial internal carotid arteries (P < .01). The contrast medium dose administered was strongly correlated with image quality (r = 0.90). CONCLUSION: Contrast medium dose is related to image quality in MR angiography of the carotid arteries performed with elliptical ordering, fluoroscopic monitoring, and manual triggering. PMID- 12755543 TI - Conversion of teaching file cases from film to digital format: a comparison between use of a diagnostic-quality digitizer and use of a flatbed scanner with transparency adapter. AB - RATIONALE AND OBJECTIVES: The authors' institution had decided to convert its radiology teaching files from film to digital media. This study was performed to determine the simplest method for converting the analog film images to digital images without a subsequent loss in diagnostic accuracy. MATERIALS AND METHODS: Twenty chest radiographs that demonstrated interstitial lung disease were randomly selected from the departmental teaching files and matched with 20 control radiographs from healthy adults. Analog film images were converted with both a diagnostic-quality film image digitizer (digitized) and a flatbed scanner equipped with a transparency adapter (scanned). Three radiology faculty members reviewed a mixed set of corresponding analog film, digitized, and scanned images. Reviewers judged whether each image depicted interstitial lung disease, indicated their level of confidence in the diagnosis, and rated each image for quality. Image quality was assessed by each reviewer subjectively at the time of viewing the individual image, without regard to other images. A one-way analysis of variance was performed to determine whether there was a statistically significant difference in diagnostic accuracy between the three image formats. Agreement in diagnosis between corresponding images in the three different formats was evaluated for each reviewer with the McNemar test. RESULTS: There was no statistically significant difference in diagnostic accuracy between analog film and scanned images, but there was such a difference between these two groups and digitized images. Accuracy was 97% for analog film, 94% for scanned, and 89% for digitized images. Results of the McNemar test showed no statistically significant difference in agreement between the analog film images and the scanned images for any of the reviewers (P > .05). CONCLUSION: A high-end flatbed scanner with transparency adapter provided accurate, simple, and inexpensive conversion of analog film teaching files to digital format, with no loss of the ability to detect or diagnose subtle abnormalities such as interstitial lung disease. PMID- 12755545 TI - What makes a great radiology resident? PMID- 12755544 TI - COMPARE radiology: creating an interactive Web-based training program for radiology with multimedia authoring software. AB - RATIONALE AND OBJECTIVES: Computer-based training has two primary benefits: Content can be presented interactively, and students can choose the time, place, and pace of learning. As a subject of medical education, radiology lends itself particularly well to computer-based training because of its highly visual content. To improve the efficiency of radiology training at their institution, the authors decided to create an interactive Web-based training site. MATERIALS AND METHODS: Working with a group of medical students knowledgeable in multimedia authoring, the authors used authoring software to create "COMPARE Radiology," an interactive training program that follows the modality-based structure of the undergraduate curriculum for radiology at the University of Erlangen-Nuremberg, Erlangen, Germany, and at medical schools worldwide. RESULTS: The Web-based program offers cases and exercises in radiographic anatomy at different selectable levels of difficulty, allowing users to test and build their knowledge of radiology. Pathologic images are initially presented without any further information. Additional information (patient history, laboratory results, reports from other imaging studies, and normal images for comparison) can be retrieved selectively and successively. Further information regarding the diagnosis and pathologic findings can be found by following links to external Web sites. The COMPARE Radiology program content is extended and updated regularly. The program is subject to internal peer review and can be evaluated by the user online. CONCLUSION: The authors' experience shows that a highly interactive Web-based training program for radiology, tailored to the requirements of the target group, can be developed economically by a team of medical students using an advanced storing system, with the guidance of a radiologist and without the help of professionally trained computer experts. PMID- 12755546 TI - Radiology residency call in the northeastern United States: comparison of difficulty and frequency in programs of different size. AB - RATIONALE AND OBJECTIVES: The purpose of this study was to develop objective measures of residency call frequency and difficulty, to establish mean values for the northeastern United States, and to test those values for correlation with program size. MATERIALS AND METHODS: A survey questionnaire was sent to 104 radiology residency programs in the northeastern United States. The programs were classified according to number of residents, as small (< 11 residents), medium sized (11-20 residents), large (21-30 residents), or very large (> or = 31 residents). The call difficulty index was defined as the number of emergency examinations per resident per year. Call frequency indexes were defined as the numbers of evenings and of nights during the 4-year residency when residents were scheduled for call. RESULTS: The average call difficulty index and standard deviation for the respondent programs was 3,855 +/- 1,779. The average call frequency index and standard deviation for evenings was 140 +/- 53 and for nights was 120 +/- 59. A significant negative correlation was found between program size on one hand and call difficulty index (r = -0.36, P = .01), evening call frequency index (r = -0.29, P = .033), and night call frequency index (r = -0.51, P < .001) on the other. Residents in small programs could expect to be on call 192 evenings and 192 nights in the 4-year residency and to perform 4,866 emergency examinations per year, as opposed to the 110 evenings and 89 nights on call and the 3,213 emergency examinations that residents in very large programs could expect. In other words, the smaller the program, the more calls residents can expect to take, and the more emergency examinations they will interpret. CONCLUSION: The mean call difficulty and off-hours call frequency indexes established for residency programs of different size in the Northeast demonstrate increasing call difficulty and increasing off-hours call frequency with decreasing program size. PMID- 12755547 TI - The ACR-RSNA Fellowship in Clinical Trials of Medical Imaging: reflections from the 2002 recipients. PMID- 12755548 TI - Slow spin exchange explains the effects of diuresis on gadolinium enhancement in MR imaging in the kidney, not in other organs. PMID- 12755549 TI - Evidence for use of coronary stents. A hierarchical bayesian meta-analysis. AB - BACKGROUND: Coronary stents are widely used in interventional cardiology, but a current quantitative systematic overview comparing routine coronary stenting with standard percutaneous transluminal coronary angioplasty (PTCA) and restricted stenting (provisional stenting) has not been published. PURPOSE: To summarize results from all randomized clinical trials comparing routine coronary stenting with standard PTCA. DATA SOURCES: Electronic databases were searched by using the key words angioplasty and stent. References from identified articles were also reviewed. In addition, several prominent general medical and cardiology journals were searched and agencies known to perform systematic reviews were consulted. STUDY SELECTION: All comparative randomized clinical trials were included, except those involving primary angioplasty for the treatment of acute myocardial infarction. DATA EXTRACTION: A specified protocol was followed, and two of the authors independently extracted the data. Outcomes assessed were total mortality, myocardial infarction, angiographic restenosis, coronary artery bypass surgery, repeated PTCA, and freedom from angina. DATA SYNTHESIS: The results were synthesized by using a Bayesian hierarchical random-effects model. A total of 29 trials involving 9918 patients were identified. There was no evidence for a difference between routine coronary stenting and standard PTCA in terms of deaths or myocardial infarctions (odds ratio, 0.90 [95% credible interval [CrI], 0.72 to 1.11]) or the need for coronary artery bypass surgery (odds ratio, 1.01 [CrI, 0.79 to 1.31]). Coronary stenting reduced the rate of restenosis (odds ratio, 0.52 [CrI, 0.37 to 0.69]) and the need for repeated PTCA (odds ratio, 0.59 [CrI, 0.50 to 0.68]). The trials showed a wide range of crossover rates from PTCA to stenting. By use of a multiplicative model, each 10% increase in crossover rate decreased the need for repeated angioplasty by approximately 8% (odds ratio multiplying factor, 1.08 [CrI, 0.98 to 1.18]). Routine stenting probably reduces the need for repeated angioplasty by fewer than 4 to 5 per 100 treated persons compared with PTCA with provisional stenting. Studies were not blinded and suggest a bias with a possible overestimation of this benefit. CONCLUSIONS: In the controlled environment of randomized clinical trials, routine coronary stenting is safe but probably not associated with important reductions in rates of mortality, acute myocardial infarction, or coronary artery bypass surgery compared with standard PTCA with provisional stenting. Coronary stenting is associated with substantial reductions in angiographic restenosis rates and the subsequent need for repeated PTCA, although this benefit may be overestimated because of trial designs. The incremental benefit of routine stenting for reducing repeated angioplasty diminishes as the crossover rate of stenting with conventional PTCA increases. PMID- 12755550 TI - A diagnostic strategy involving a quantitative latex D-dimer assay reliably excludes deep venous thrombosis. AB - BACKGROUND: Because clinical diagnosis is inaccurate, objective testing is usually considered necessary when patients present with suspected deep venous thrombosis (DVT). OBJECTIVE: To determine whether a negative result on a quantitative latex D dimer assay eliminates the need for further investigation in patients with a low or moderate pretest probability of DVT. DESIGN: Prospective cohort study. SETTING: Three tertiary care hospitals in Canada. PATIENTS: 556 consecutive outpatients with suspected first DVT. INTERVENTION: Patients were categorized as having a low, moderate, or high pretest probability of DVT and then underwent D-dimer testing. Patients with low or moderate pretest probability and a negative D-dimer result had no further diagnostic testing and received no anticoagulant therapy. Serial compression ultrasonography was performed in all other patients. Patients who did not receive a diagnosis of DVT were followed for symptomatic venous thromboembolism. MEASUREMENTS: Objectively confirmed symptomatic venous thromboembolic events during 3 months of follow-up. RESULTS: 283 patients (51%) had low or moderate pretest probability and a negative D-dimer result. One of these patients had DVT during follow-up (negative likelihood ratio, 0.05 [CI, 0.01 to 0.23]). The negative likelihood ratio of the d -dimer test in all patients was 0.03 (CI, 0.01 to 0.16). CONCLUSION: A negative result on a quantitative latex d -dimer assay safely eliminates the need for further testing in patients with low or moderate pretest probability of DVT. PMID- 12755552 TI - Case reports of heart failure after therapy with a tumor necrosis factor antagonist. AB - BACKGROUND: Etanercept and infliximab are U.S. Food and Drug Administration approved tumor necrosis factor (TNF) antagonists. OBJECTIVE: To describe adverse event reports of heart failure after TNF antagonist therapy. DESIGN: Case series. SETTING: The U.S. Food and Drug Administration's MedWatch program. PATIENTS: 47 patients who developed new or worsening heart failure during TNF antagonist therapy. MEASUREMENTS: Clinical and laboratory reports. RESULTS: After TNF antagonist therapy, 38 patients developed new-onset heart failure and 9 patients experienced heart failure exacerbation. Of the 38 patients with new-onset heart failure, 19 (50%) had no identifiable risk factors. Ten patients younger than 50 years of age developed new-onset heart failure after receiving TNF antagonists. After TNF antagonist therapy was discontinued and heart failure therapy was started in these 10 patients, 3 had complete resolution of heart failure, 6 improved, and 1 died. CONCLUSION: In a fraction of patients, TNF antagonists might induce new-onset heart failure or exacerbate existing disease. PMID- 12755551 TI - The cost-effectiveness of cyclooxygenase-2 selective inhibitors in the management of chronic arthritis. AB - BACKGROUND: Rofecoxib and celecoxib (coxibs) effectively treat chronic arthritis pain and reduce ulcer complications by 50% compared with nonselective nonsteroidal anti-inflammatory drugs (NSAIDs). However, their absolute risk reduction is small and the cost-effectiveness of treatment is uncertain. OBJECTIVE: To determine whether the degree of risk reduction in gastrointestinal complications by coxibs offsets their increased cost compared with a generic nonselective NSAID. DESIGN: Cost-utility analysis. DATA SOURCES: Systematic review of MEDLINE and published abstracts. TARGET POPULATION: Patients with osteoarthritis or rheumatoid arthritis who are not taking aspirin and who require long-term NSAID therapy for moderate to severe arthritis pain. PERSPECTIVE: Third party payer. INTERVENTIONS: Naproxen, 500 mg twice daily, and coxib, once daily. Patients intolerant of naproxen were switched to a coxib. TIME HORIZON: Lifetime. OUTCOME MEASURES: Incremental cost per quality-adjusted life-year (QALY) gained. RESULTS OF BASE-CASE ANALYSIS: Using a coxib instead of a nonselective NSAID in average-risk patients cost an incremental 275 809 dollars per year to gain 1 additional QALY. RESULTS OF SENSITIVITY ANALYSIS: The incremental cost per QALY gained decreased to 55 803 dollars when the analysis was limited to the subset of patients with a history of bleeding ulcers. The coxib strategy became dominant when the cost of coxibs was reduced by 90% of the current average wholesale price. In probabilistic sensitivity analysis, if a third-party payer was willing to pay 150 000 dollars per QALY gained, then 4.3% of average-risk patients would fall within the budget. CONCLUSIONS: The risk reduction seen with coxibs does not offset their increased costs compared with nonselective NSAIDs in the management of average-risk patients with chronic arthritis. However, coxibs may provide an acceptable incremental cost-effectiveness ratio in the subgroup of patients with a history of bleeding ulcers. PMID- 12755553 TI - Advance care planning for fatal chronic illness: avoiding commonplace errors and unwarranted suffering. AB - Patients with eventually fatal illnesses often receive routine treatments in response to health problems rather than treatments arising from planning that incorporates the patient's situation and preferences. This paper considers the case of an elderly man with advanced lung disease who had mechanical ventilation and aggressive intensive care, in part because his nursing home clinicians did not complete an advance care plan and his do-not-resuscitate order did not accompany him to the hospital. The errors that led to his hospitalization and his unwanted treatment there demonstrate how the ordinary lack of advance care planning is deleterious for patients who are nearing the end of life. We discuss serious, recurring, and generally unnoticed errors in planning for care near the end of life and possible steps toward improvement. Repairing these shortcomings will require quality improvement and system redesign efforts, methods familiar from patient safety initiatives. Reliable improvement will also require making it unacceptable for clinicians to fail to plan ahead for care during fatal chronic illness. PMID- 12755554 TI - Philadelphia chromosome-positive leukemias: from basic mechanisms to molecular therapeutics. AB - The Philadelphia chromosome translocation (t(9;22)) results in the molecular juxtaposition of two genes, BCR and ABL, to form an aberrant BCR-ABL gene on chromosome 22. BCR-ABL is critical to the pathogenesis of chronic myelogenous leukemia and a subset of acute leukemias. The chimeric Bcr-Abl protein has constitutively elevated tyrosine phosphokinase activity. This abnormal enzymatic activation is critical to the oncogenic potential of Bcr-Abl. Initially, protein kinases were thought to be poor therapeutic targets because of their ubiquitous nature and crucial role in many normal physiologic processes. However, the advent of imatinib mesylate (Gleevec, Novartis Pharmaceuticals, Basel, Switzerland), formerly known as STI571 and CGP57148B, demonstrated that designer kinase inhibitors could be specific. This agent has shown striking activity in chronic myelogenous leukemia. It also inhibits phosphorylation of Kit (stem-cell factor receptor) and platelet-derived growth factor receptor. In addition, it has shown similar impressive responses, with little host toxicity, in gastrointestinal stromal tumors, which harbor activating Kit mutations, and in tumors with activated platelet-derived growth factor receptor. The studies of imatinib mesylate provide proof-of-principle for using aberrant kinases as a therapeutic target and are a model for the promise of molecular therapeutics. This paper reviews the current knowledge on the function of Bcr-Abl and its normal counterparts (Bcr and Abl), as well as the impact of this knowledge on the development of a remarkably successful targeted therapy approach. PMID- 12755555 TI - Lessons from the Stroke Prevention in Atrial Fibrillation trials. AB - Atrial fibrillation predisposes to left atrial thrombus formation and carries a sixfold increased risk for stroke. Antithrombotic therapies are the mainstay for stroke prevention. The National Institute of Neurological Disorders and Stroke sponsored Stroke Prevention in Atrial Fibrillation (SPAF) studies assessed the value of warfarin, aspirin, and their combination for preventing stroke in six multicenter trials involving 3950 participants. This review presents the major results and implications, which offer unique perspectives on antithrombotic therapies for stroke prevention in atrial fibrillation. Warfarin and aspirin reduce stroke. Anticoagulation substantially benefits high-risk patients with atrial fibrillation, while many younger patients with atrial fibrillation have a low stroke rate when given aspirin. Pathogenetic and transesophageal echocardiographic correlations shed light on mechanisms by which antithrombotic agents prevent stroke. Warfarin inhibits formation of atrial appendage thrombi and markedly reduces cardioembolic strokes, while aspirin primarily prevents smaller, noncardioembolic strokes. The SPAF III stroke risk stratification scheme has been validated for identifying patients with high versus moderate versus low risk for stroke. Women with atrial fibrillation benefit from anticoagulation significantly more than men do. Many elderly patients with recurrent paroxysmal atrial fibrillation have high rates of stroke. Antithrombotic prophylaxis should be individualized on the basis of the estimated risk for stroke during aspirin therapy and the risk for bleeding during anticoagulation. Overall, nearly one third of patients with atrial fibrillation are low risk and should be treated with aspirin, and about one third are high risk and should receive warfarin if it can be given safely. For patients at moderate risk for stroke, patient preferences and access to reliable anticoagulation monitoring are particularly relevant. PMID- 12755556 TI - Medical professionalism in the new millennium: a physician charter 15 months later. PMID- 12755557 TI - Why have stents replaced balloons? Underwhelming evidence. PMID- 12755558 TI - The Charter on Medical Professionalism and the limits of medical power. PMID- 12755559 TI - Past and present. PMID- 12755560 TI - Any oasis will do. PMID- 12755562 TI - Charter on medical professionalism: putting the charter into practice. PMID- 12755564 TI - Charter on medical professionalism: putting the charter into practice. PMID- 12755565 TI - Charter on medical professionalism: putting the charter into practice. PMID- 12755567 TI - Charter on medical professionalism: putting the charter into practice. PMID- 12755568 TI - Charter on medical professionalism: putting the charter into practice. PMID- 12755570 TI - Charter on medical professionalism: putting the charter into practice. PMID- 12755572 TI - Charter on medical professionalism: putting the charter into practice. PMID- 12755574 TI - Charter on medical professionalism: putting the charter into practice. PMID- 12755575 TI - Charter on medical professionalism: putting the charter into practice. PMID- 12755580 TI - The vaccine crossroads. PMID- 12755581 TI - Summaries for patients. Tumor necrosis factor antagonists and heart failure. PMID- 12755583 TI - Summaries for patients. Negative results on a quantitative latex D-dimer test exclude deep leg clots. PMID- 12755582 TI - Summaries for patients. The cost-effectiveness of cyclooxygenase-2 inhibitors for treating chronic arthritis. PMID- 12755584 TI - Summaries for patients. Benefits of coronary stents. PMID- 12755585 TI - Nitric oxide-induced interstrand cross-links in DNA. AB - The DNA damaging effects of nitrous acid have been extensively studied, and the formation of interstrand cross-links have been observed. The potential for this cross-linking to occur through a common nitrosating intermediate derived from nitric oxide is investigated here. Using a HPLC laser-induced fluorescence (LIF) system, the amount of interstrand cross-link formed on nitric oxide treatment of the 5'-fluorescein-labeled oligomer ATATCGATCGATAT was determined. This self complimentary sequence contains two 5'-CG sequences, which is the preferred site for nitrous acid-induced cross-linking. Nitric oxide was delivered to an 0.5 mM oligomer solution at 15 nmol/mL/min to give a final nitrite concentration of 652 microM. The resulting concentration of the deamination product, xanthine, in this sample was found to be 211 +/- 39 nM, using GC/MS, and the amount of interstrand cross-link was determined to be 13 +/- 2.5 nM. Therefore, upon nitric oxide treatment, the cross-link is found at approximately 6% of the amount of the deamination product. Using this system, detection of the cross-link is also possible for significantly lower doses of nitric oxide, as demonstrated by treatment of the same oligomer with NO at a rate of 18 nmol/mL/min resulting in a final nitrite concentration of 126 microM. The concentration of interstrand cross link was determined to be 3.6 +/- 0.1 nM in this sample. Therefore, using the same dose rate, when the total nitric oxide concentration delivered drops by a factor of approximately 5, the concentration of cross-link drops by a factor of about 4-indicating a qausi-linear response. It may now be possible to predict the number of cross-links in a small genome based on the number of CpG sequences and the yield of xanthine derived from nitrosative deamination. PMID- 12755586 TI - Structure elucidation, synthesis, and contact allergenic activity of a major hydroperoxide formed at autoxidation of the ethoxylated surfactant C12E5. AB - Ethoxylated alcohols, widely used as surfactants, are known to be susceptible to oxidation when exposed to air. At autoxidation, a complex mixture is formed, in which alkyl poly(ethylene glycol) aldehydes, alkyl poly(ethylene glycol) formates, hydroxyaldehydes, and formaldehyde have previously been identified. These compounds are all secondary oxidation products, some of which have been shown to be skin sensitizers and irritants. The primary oxidation products from ethoxylated alcohols are described in the literature as peroxides and hydroperoxides, but their structures have not been elucidated more closely. Peroxides and hydroperoxides are usually reactive species and can be suspected to be biologically active as skin sensitizers and irritants. In the present investigation, we studied the autoxidation of the pure ethoxylated alcohol pentaethylene glycol mono-n-dodecyl ether (C(12)E(5)), using NMR and HPLC-MS. On the basis of experience from previous studies on a small model compound, diethyleneglycol monoethyl ether (C(2)E(2)), the hydroperoxide expected to be found in the highest amount in autoxidized C(12)E(5) was synthesized and used as a reference substance in the analyses. This same hydroperoxide, 16-hydroperoxy 3,6,9,12,15-pentaoxaheptacosan-1-ol, was identified in the autoxidation mixture of C(12)E(5), and its sensitizing capacity was determined. It was found to be a moderate allergen in experimental sensitization studies in guinea pigs. Our data further indicate the presence of at least three additional hydroperoxides in the autoxidation mixture of C(12)E(5), one of which was identified as 1-hydroperoxy 3,6,9,12,15-pentaoxaheptacosan-1-ol. The results accentuate the importance of controlling the storage, transportation, and handling conditions of ethoxylated surfactants, to avoid the formation of allergenic and skin irritant oxidation products. PMID- 12755587 TI - Selective covalent binding of acrylonitrile to Cys 186 in rat liver carbonic anhydrase III in vivo. AB - Covalent binding of reactive chemical species to tissue proteins is a common, but poorly understood, mechanism of toxicity. Identification of the proteins and the specific amino acid residues within the proteins that are chemically modified will aid our understanding of the toxification/detoxification mechanisms involved in covalent binding. Acrylonitrile (AN) is a commercial vinyl monomer that is acutely toxic and readily binds to tissue proteins. Total covalent binding of AN to tissue proteins is highly correlated with acute toxicity. Two-dimensional PAGE and autoradiography were used to locate proteins in male rat liver cytosol that are radiolabeled following administration of [2,3-(14)C]AN in vivo. Four intensely labeled spots were prominent in the autoradiogram and formed an apparent "charge-train" at approximately 30 kDa. Tryptic peptide mapping by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS was used to identify all of the spots as carbonic anhydrase III (CAIII). HPLC of the tryptic digests combined with MALDI-TOF MS was used to localize the radiolabel to tryptic fragment T22 containing amino acids 171-187. This tryptic fragment contains two Cys residues (Cys181 and Cys186) in the rat CAIII sequence. Electrospray ionization ion-trap MS was used to sequence the peptide and establish that only Cys186 was labeled. Thus, although AN is considered to be highly reactive, our data indicate that it does not react indiscriminately with rat CAIII but rather is selective for one out of five Cys residues. Rat liver CAIII has previously been shown to protect cells against oxidative stress. Our data suggest that CAIII is also capable of scavenging reactive xenobiotics and may help prevent covalent binding to more critical macromolecules. PMID- 12755588 TI - Conformations of benzene- and dibenzo[a,l]pyrene diol epoxides studied by density functional theory: ground states, transition states, dynamics, and solvent effects. AB - The (-)-anti- and (+)-syn-diol epoxides of dibenzo[a,l]pyrene (DBPDE, 11,12 dihydroxy-13,14-epoxy-11,12,13,14-tetrahydrodibenzo[a,l]pyrene) and the stereochemically corresponding benzene diol epoxides (BDE, 1,2-dihydroxy-3,4 epoxy-1,2,3,4-tetrahydrobenzene) have been studied by density functional theory (DFT) to determine the structures, energies, dynamics, thermal properties, and solvent effects on the different conformers. The smaller BDE is used as a model compound for studies of transitions between diequatorial and diaxial conformations of the hydroxyl groups. It was found that DBPDE is distorted due to overcrowding in the fjord region and that the arene oxide prefers to be on the same side of the saturated ring as the distal ring ("in") in most stereoisomeric states. For the anti-diastereomer, a diequatorial orientation of the hydroxyl groups is preferred, while the orientation preference in the syn-diastereomer seems to depend on the solvent and the in/out conformation. Transition states for the interconversions between in and out conformations of DBPDE as well as between diequatorial and diaxial conformations on BDE have been found, and transition rates have been estimated by transition state theory. The barriers are found to be moderate, the highest being 9.6 kcal/mol. Solvent effects as well as zero point vibrational energy and thermal effects were included and found to be significant in some cases. The results presented here are in agreement with previous experimental studies. PMID- 12755589 TI - An integrated approach to identifying chemically induced posttranslational modifications using comparative MALDI-MS and targeted HPLC-ESI-MS/MS. AB - Identification of multiple and novel posttranslational modifications remains a major challenge in proteomics. The present approach uses comparative analysis by matrix-assisted laser/desorption ionization (MALDI) MS of proteolytic digests from control and treated proteins to target differences due to modifications, without initial assumption as to type or residue localization. Differences between modified and unmodified digest MS spectra highlight peptides of interest for subsequent tandem mass spectrometry (MS/MS) analysis. Targeted HPLC electrospray ionization (ESI)-MS/MS is then used to fragment peptides, and manual de novo sequencing is used to determine the amino acid sequence and type of modification. This strategy for identifying posttranslational modifications in an unbiased manner is particularly useful for finding modifications produced by exogenous chemicals. Successful characterization of chemically induced posttranslational modifications and novel chemical adducts is given as an example of the use of this strategy. Histone H4 from butyrate-treated LLC-PK1 cells is separated on a gel into bands representing different overall charge state. Bands are analyzed by comparative MALDI-MS and LC-MS/MS to identify the sites of methylation and acetylation. Previous attempts to identify chemically adducted proteins in vivo have been unsuccessful in part due to a lack of understanding of the final adduct form. Cytochrome c is adducted in vitro with benzoquinone, an electrophilic metabolite of benzene capable of interacting with nucleophilic sites within proteins. De novo sequencing identifies a novel cyclized diquinone adduct species as the major reaction product, targeting Lys and His residues at two specific locations on the protein surface. This unpredicted reaction product is characterized using our unbiased methods for detection and demonstrates the important influence of protein structure on chemical adduction. PMID- 12755590 TI - Allyl alcohol activation of protein kinase C delta leads to cytotoxicity of rat hepatocytes. AB - Hepatotoxicity of allyl alcohol involves its bioactivation to acrolein and subsequent protein sulfhydryl loss and lipid peroxidation. However, the links between these events and hepatocellular death are not known. The purpose of these studies was to examine whether specific signal transduction pathways are associated with allyl alcohol toxicity in hepatocytes. Inhibition or augmentation of cyclic AMP and/or protein kinase A (PKA) by Rp-Ado-3N,5N-cyclic monophosphorothioate triethylamine salt or 3-isobutyl-1-methylxanthine had no effect on allyl alcohol-induced cell death. H-7, an inhibitor of PKA, PKC, and PKG, partially inhibited cell killing by allyl alcohol, whereas chelerythrine chloride, a nonselective PKC inhibitor, almost completely abolished allyl alcohol cytotoxicity. Neither 2,2N,3,3N,4,4N-hexahydroxy-1,1N,-biphenyl-6,6N-dimethanol dimethyl ether, a selective PKC alpha and beta inhibitor, nor bisindolylmaleimide I, an inhibitor of PKC alpha, beta, and epsilon, had any effect on allyl alcohol cytotoxicity. In contrast, rottlerin, a selective PKCdelta inhibitor, blocked hepatocellular killing by allyl alcohol. Cytoprotection by chelerythrine chloride and rottlerin was not the result of inhibition of bioactivation of allyl alcohol because each inhibitor also prevented cell death from acrolein. Western blotting and immunohistochemical techniques revealed that allyl alcohol stimulated phosphorylation and translocation of PKCdelta to hepatocyte membranes (i.e., activation), and this activity was inhibited by rottlerin. Cell death appeared to occur via oncotic necrosis rather than apoptosis based on single-stranded DNA ELISA and propidium iodide staining. Together, these results indicate that activation of PKCdelta is a critical, early event in initiating hepatocyte injury and death from allyl alcohol. PMID- 12755591 TI - Identification of adducts formed by pyridyloxobutylation of deoxyguanosine and DNA by 4-(acetoxymethylnitrosamino)-1-(3-pyridyl)-1-butanone, a chemically activated form of tobacco specific carcinogens. AB - The tobacco specific carcinogens 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and N'-nitrosonornicotine (NNN) are metabolically activated to 4-oxo-4-(3 pyridyl)-1-butanediazohydroxide (7), which is known to pyridyloxobutylate DNA. A substantial proportion of the adducts in this DNA releases 4-hydroxy-1-(3 pyridyl)-1-butanone (HPB, 11) under various hydrolysis conditions, including neutral thermal hydrolysis. These HPB-releasing DNA adducts have been detected in target tissues of animals treated with NNK and NNN as well as in lung tissue from smokers. Although their presence in pyridyloxobutylated DNA was conclusively demonstrated 15 years ago, their structures have not been previously determined. We investigated this question in the present study by determining the structures of products formed in reactions with dGuo and DNA of 4-(acetoxymethylnitrosamino) 1-(3-pyridyl)-1-butanone (NNKCH(2)OAc, 3), a stable precursor to 7. Reaction mixtures from NNKCH(2)OAc and dGuo were analyzed by liquid chromatography electrospray ionization-mass spectrometry (LC-ESI-MS) with selected ion monitoring at m/z 415. A major peak was detected and identified as 7-[4-oxo-4-(3 pyridyl)but-1-yl]dGuo (37) by its ESI-MS fragmentation pattern and by neutral thermal hydrolysis, which converted it to 11 and 7-[4-oxo-4-(3-pyridyl)but-1 yl]Gua (26). The latter was identified by comparison to synthetic 26 using LC-ESI MS with selected ion monitoring at m/z 299, M + 1 of 26. Further evidence was obtained by NaBH(4) reduction of 26 to 7-[4-hydroxy-4-(3-pyridyl)but-1-yl]Gua, which was also matched with a standard. Adduct 37 was similarly identified in enzyme hydrolysates of DNA reacted with NNKCH(2)OAc, accounting for 30-35% of the HPB-releasing adducts in this DNA. Several other adducts resulting from pyridyloxobutylation of the N(2)- and O(6)-positions of Gua were also identified as products in the dGuo or DNA reactions by comparison to standards; their concentrations were considerably less than that of 37. These adducts were N(2)-[4 oxo-4-(3-pyridyl)but-1-yl]dGuo (23), N(2)-[4-oxo-4-(3-pyridyl)but-2-yl]dGuo (25), N(2)-[2-(3-pyridyl)tetrahydrofuran-2-yl]dGuo (31a) (or its open chain tautomer 31b), and O(6)-[4-oxo-4-(3-pyridyl)but-1-yl]dGuo (10). Adducts 23, 25, and 10 did not release HPB upon neutral thermal hydrolysis. The results of this study provide the first structural identification of an HPB-releasing DNA adduct of the tobacco specific nitrosamines NNK and NNN. PMID- 12755592 TI - Studies of chemical selectivity of hapten, reactivity, and skin sensitization potency. 3. Synthesis and studies on the reactivity toward model nucleophiles of the 13C-labeled skin sensitizers, 5-chloro-2-methylisothiazol-3-one (MCI) and 2 methylisothiazol-3-one (MI). AB - The skin sensitizers, 5-chloro-2-methylisothiazol-3-one (MCI) and 2 methylisothiazol-3-one (MI), have been synthesized isotopically labeled with (13)C at all carbon positions. The reactivity of 3-[(13)C]-, 4-[(13)C]-, and 5 [(13)C]MCI and MI toward a series of model nucleophiles for protein amino acid residues, i.e., butylamine, imidazole, sodium propanethiolate, and sodium phenoxide, was followed by (13)C and (1)H[(13)C] NMR spectroscopy. While MCI was found to react quantitatively with sodium propanethiolate and butylamine and significantly with imidazole and sodium phenoxide, MI reacted only with sodium propanethiolate. Reaction of MCI with nonthiol nucleophiles proceeded through an initial addition-elimination at position 5, leading to stable substitution adducts in the case of imidazole and sodium phenoxide. In the case of butylamine, the initial adduct was subjected to extra reactions at the sulfur atom through a cleavage of the S-N bond, leading to open adducts of the thioamide or amide type. Experiments carried out with N-acetyl-Cys, in excess or in deficiency, indicated that thiol nucleophiles reacted first at the sulfur atom through a cleavage of the S-N bond followed by extra nucleophilic reactions leading to open adducts of the mercaptothioester or mercaptoester type. Reaction of MCI with thiol nucleophiles gave products consistent with the formation of a reactive thioacyl chloride intermediate able to react with other nucleophiles present in the reaction medium. As a consequence, N-acetyl-Cys was found to be able to activate MCI toward N(alpha)-acetyl-Lys under physiological conditions to form adducts of the thioamide or amide type. Thus MCI, a strong sensitizer, and MI, a weak sensitizer, were found to react with different nucleophiles through different mechanisms. Although both MCI and MI can react with thiol nucleophiles, only MCI is capable of significantly reacting with amino nucleophiles of the Lys or His type. Moreover, MCI could be activated by a prior reaction with thiols. PMID- 12755594 TI - Photosensitization of DNA strand breaks by three phenothiazine derivatives. AB - The interaction and the photosensitizing activity of three phenothiazine derivatives, fluphenazine hydrochloride (FP), thioridazine hydrochloride (TR), and perphenazine (PP), toward DNA were studied. Evidences obtained from various spectroscopic studies such as fluorimetric and linear dichroism measurements indicate that these derivatives bind to the DNA at least in two ways: intercalation and external stacking on the DNA helix, depending on their relative concentrations. Irradiation of supercoiled plasmid DNA in the presence of these phenothiazines leads to single strand breaks. The DNA photocleavage appears to be due to externally bound molecules rather than to those intercalated. The highest photocleavage activity was observed with PP and TR whereas FP was less efficient. The efficiency of the photocleavage in aerated and deaerated solutions does not change thus indicating that an involvement of singlet oxygen can be excluded. Primer extension analysis of plasmid DNA irradiated in the presence of phenothiazines indicates that photocleavage of DNA occurs predominantly at Gua and Cyt residues. Laser flash experiments carried out in the presence of 2' deoxyguanosine 5'-monophosphate reveal an efficient electron transfer between the nucleotide and the radical cations produced by photoionization of the phenothiazines. In the presence of DNA, an electron transfer process takes place within the laser pulse from the lowest singlet state of phenothiazines to the DNA bases; the time-resolved measurements showed that the back-electron transfer is a negligible decay pathway for the charged species. PMID- 12755593 TI - Synthesis and characterization of peptides containing a cyclic Val adduct of diepoxybutane, a possible biomarker of human exposure to butadiene. AB - 1,3-Butadiene, a potential human carcinogen widely used in industry, is oxidized by cytochrome P450 to diepoxybutane (DEB), which is the most mutagenic of the known butadiene metabolites. Assessment of the toxicological significance of DEB formation in humans and animals requires identification of a biomarker uniquely associated with DEB for use in molecular dosimetry studies. We wished to develop a specific and sensitive assay for one such suitable marker, the cyclic adduct 2 (3,4-dihydroxypyrrolidin-1-yl)-3-methylbutyramide (pyr-V), which is formed from addition of DEB to the terminal Val of the alpha- and beta-chains of hemoglobin. We needed to prepare a pure, rigorously characterized DEB-modified N-terminal oligopeptide for raising antibodies both to use in an immunoaffinity purification step and to standardize the assay. In addition, we needed a pure isotopomer to serve as an internal standard for quantitation by LC-MS. Direct modification of the globin sequences by reaction with DEB in vitro proved to be unproductive. We therefore opted to synthesize the cyclic Val adduct and incorporate it by FMOC chemistry into the appropriate oligopeptide sequences. In vitro and in vivo, butadiene is oxidized to enantiomeric and meso forms of DEB. A priori, all three DEB isomers are expected to form pyr-V adducts, resulting in three diastereomeric N-terminal peptides. We therefore synthesized a mixture of the cyclic Val diastereomers as their methyl esters by reaction of DEB with l-Val methyl ester hydrochloride. After protection as the di-O-tert-butyl derivatives, the mixture of 2-(3,4-di-t-butoxypyrrolidin-1-yl)-3-methylbutyric acid diastereomers was incorporated as the N-terminal residue into the 1-11 human globin alpha-chain sequence VLSPADKTNVK. The presence of the three diastereomers was confirmed by two-dimensional correlation NMR spectroscopy and temperature-dependent (1)H NMR. This strategy enabled us to obtain pure, rigorously characterized haptens in quantity for the preparation of polyclonal antibodies. Use of FMOC-protected (2)H(3)-Leu in the automated oligopeptide synthesis provided the required isotopomers for use as internal standard. PMID- 12755595 TI - Tryptophan-14 is the preferred site of DBNBS spin trapping in the self peroxidation reaction of sperm whale metmyoglobin with a single equivalent of hydrogen peroxide. AB - The 3,5-dibromo-4-nitrosobenzenesulfonate (DBNBS)-metmyoglobin adduct formed following the horse metmyoglobin-H(2)O(2) reaction has been assigned to both a tyrosyl and a tryptophanyl residue radical. At low H(2)O(2), hyperfine coupling to a (13)C atom in sperm whale metmyoglobin labeled at the tryptophan residues with (13)C allowed the unequivocal assignment of the primary adduct to a tryptophanyl radical. Trapping at Trp-14 of sperm whale myoglobin was indicated by greatly decreased electron paramagnetic resonance (EPR) spectral intensity of the DBNBS adducts of the Trp-14-Phe recombinant proteins. Complex EPR spectra with partially resolved hyperfine splittings from several atoms were obtained by pronase treatment of the DBNBS/*W14F metmyoglobin adducts. The EPR spectra of authentic DBNBS/*Tyr adducts were incubation time-dependent; the late time spectra resembled the spectra of pronase-treated DBNBS/*W14F sperm whale myoglobin adducts, suggesting formation of an unstable tyrosyl radical adduct in the latter proteins. When the H(2)O(2):metmyoglobin ratio was increased to 5:1, the EPR spectrum after pronase treatment supported trapping of a tyrosyl radical, although similar decreases in tryptophan content were detected at H(2)O(2):metmyoglobin ratios of 1:1 and 5:1. PMID- 12755596 TI - Nucleophilic reactions between thiols and a tobacco specific nitrosamine metabolite, 4-hydroxy-1-(3-pyridyl)-1-butanone. AB - 4-Hydroxy-1-(3-pyridyl)-1-butanone (HPB) is a metabolite of the tobacco specific nitrosamines, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and N' nitrosonornicotine (NNN). HPB is also a breakdown product of covalently bound pyridyloxobutyl adducts resulting from NNK and NNN exposure. HPB released from DNA or hemoglobin has been used as an important dosimeter of tobacco specific nitrosamine exposure in a variety of studies. This compound is not reactive with cellular nucleophiles under biological conditions. We have discovered that HPB reacts with nucleophiles under acidic conditions to form cyclic tetrahydrofuranyl reaction products. Dithiothreitol, 2-mercaptoethanol, and N-acetylcysteine all reacted with HPB under these reaction conditions. In addition, reactions were observed with buffer chemicals such as 4-(2-hydroxyethyl)-1 piperazineethanesulfonic acid and tris(hydroxymethyl)aminomethane. The resulting cyclic adducts were unstable at room temperature. Their half-lives were significantly longer under neutral conditions than under acidic conditions. NMR studies established that the cyclic form of HPB, 2-hydroxy-2-(3-pyridyl)-2,3,4,5 THF, is present at significant concentrations in acidic solutions. The observation of this cyclic compound suggests that the reaction with nucleophiles may occur via a cyclic oxonium ion intermediate. This reaction was significant in our biological samples; there was up to 40% conversion of [5-(3)H]HPB to cyclic DTT-derived compounds when acidic DNA repair reactions containing [5 (3)H]pyridyloxobutylated DNA were stored overnight at -20 degrees C. Therefore, long-term storage of acid hydrolysates of pyridyloxobutylated DNA or protein for the analysis of HPB-releasing adducts could result in an underestimation of HPB releasing adduct in those samples. In addition, these observations provide a mild synthetic method to prepare large quantities of cyclic 2-(3-pyridyl)-2,3,4,5-THF adducts predicted to result from pyridyloxobutylation of important cellular nucleophiles as a result of NNK and/or NNN exposure. PMID- 12755597 TI - Catechol estrogen 4-hydroxyequilenin is a substrate and an inhibitor of catechol O-methyltransferase. AB - Redox and/or electrophilic metabolites formed during estrogen metabolism may play a role in estrogen carcinogenesis. 4-Hydroxyequilenin (4-OHEN) is the major phase I catechol metabolite of the equine estrogens equilenin and equilin, which are components of the most widely prescribed estrogen replacement formulation, Premarin. Previously, we have found that 4-OHEN rapidly autoxidized to an o quinone in vitro and caused toxic effects such as the inactivation of human detoxification enzymes. 4-OHEN has also been shown to be a substrate for catechol O-methyltransferase (COMT) in human breast cancer cells. In the present study, we demonstrated that 4-OHEN was not only a substrate of recombinant human soluble COMT in vitro with a K(m) of 2.4 microM and k(cat) of 6.0 min(-)(1) but it also inhibited its own methylation by COMT at higher concentrations in the presence of the reducing agent dithiothreitol. In addition, 4-OHEN was found to be an irreversible inhibitor of COMT-catalyzed methylation of the endogenous catechol estrogen 4-hydroxyestradiol with a K(i) of 26.0 microM and a k(2) of 1.62 x 10( )(2) s(-)(1). 4-OHEN in vitro not only caused the formation of intermolecular disulfide bonds as demonstrated by gel electrophoresis, but electrospray ionization mass spectrometry and matrix-assisted laser desorption ionization time of-flight mass spectrometry also showed that 4-OHEN alkylated multiple residues of COMT. Peptide mapping experiments further indicated that Cys33 in recombinant human soluble COMT was the residue most likely modified by 4-OHEN in vitro. These data suggest that inhibition of COMT methylation by 4-OHEN might reduce endogenous catechol estrogen clearance in vivo and further enhance toxicity. PMID- 12755598 TI - Structure of frataxin iron cores: an X-ray absorption spectroscopic study. AB - X-ray absorption spectroscopy at the iron K-edge indicates that the iron cores of human and yeast frataxin polymers assembled in vitro are identical to each other and are similar but not identical to ferritin cores. Both frataxin polymers contain ferrihydrite, a biomineral composed of ferric oxide/hydroxide octahedra. The ferrihydrite in frataxin is less ordered than iron cores of horse spleen ferritin, having fewer face-sharing Fe-Fe interactions but similar double corner sharing interactions. The extended X-ray absorption fine structure (EXAFS) analysis agrees with previous electron microscopy data showing that frataxin cores are composed of very small ferrihydrite crystallites. PMID- 12755599 TI - BAFF/BLyS receptor 3 comprises a minimal TNF receptor-like module that encodes a highly focused ligand-binding site. AB - BAFF/BLyS, a member of the tumor necrosis family (TNF) superfamily of ligands, is a crucial survival factor for B cells. BAFF binds three receptors, TACI, BCMA, and BR3, with signaling through BR3 being essential for promoting B cell function. Typical TNF receptor (TNFR) family members bind their cognate ligands through interactions with two cysteine-rich domains (CRDs). However, the extracellular domain (ECD) of BR3 consists of only a partial CRD, with cysteine spacing distinct from other modules described previously. Herein, we report the solution structure of the BR3 ECD. A core region of only 19 residues adopts a stable structure in solution. The BR3 fold is analogous to the first half of a canonical TNFR CRD but is stabilized by an additional noncanonical disulfide bond. BAFF-binding determinants were identified by shotgun alanine-scanning mutagenesis of the BR3 ECD expressed on phage. Several of the key BAFF-binding residues are presented from a beta-turn that we have shown previously to be sufficient for ligand binding when transferred to a structured beta-hairpin scaffold [Kayagaki, N., Yan, M., Seshasayee, D., Wang, H., Lee, W., French, D. M., Grewal, I. S., Cochran, A. G., Gordon, N. C., Yin, J., Starovasnik, M. A, and Dixit, V. M. (2002) Immunity 10, 515-524]. Outside of the turn, mutagenesis identifies additional hydrophobic contacts that enhance the BAFF-BR3 interaction. The crystal structure of the minimal hairpin peptide, bhpBR3, in complex with BAFF reveals intimate packing of the six-residue BR3 turn into a cavity on the ligand surface. Thus, BR3 binds BAFF through a highly focused interaction site, unprecedented in the TNFR family. PMID- 12755600 TI - Sequence-selective targeting of long stretches of the DNA minor groove by a novel dimeric bis-benzimidazole. AB - A dimeric bis-benzimidazole molecule has been designed by computer modeling to bind to a DNA sequence via the DNA minor groove that covers a complete turn of B DNA. A series of bis-benzimidazole dimers incorporating a -O-(CH(2))(n)()-X CH(2))(n)()-O- linker, with n = 2 or 3 and X = O or N(+)H(Me), were screened for their capacity to fit the DNA minor groove. The modeling studies enabled an optimal linker to be devised (n = 3, X = N(+)H(Me)), and the synthesis of the predicted "best" molecule, N-methyl-N,N-bis-3,3-[4'-[5' '-(2' "-p-methoxyphenyl) 5' "-1H-benzimidazolyl]-2' '-1H-benzimidazolyl]phenoxypropylamine (5), is reported. The optimized linker permits the two symmetric bis-benzimidazole motifs to maintain hydrogen-bonded contacts with the floor of the DNA minor groove. DNase I footprinting studies have shown that this ligand binds with high affinity to sequences representing approximately a complete turn of B-DNA, represented by the [A.T](4)-[G.C]-[A.T](4) motif, and only poorly to sequences of half this site size, in accord with the computer modeling studies. Compound 5 does not show acute cellular cytotoxicity, in contrast with its monomeric bis-benzimidazole precursors, yet is rapidly taken up into cells. PMID- 12755601 TI - Investigating the role of the latch in the positive supercoiling mechanism of reverse gyrase. AB - Reverse gyrase is the only topoisomerase known to positively supercoil DNA and the only protein unique to hyperthermophiles. The enzyme comprises an N-terminal ATPase domain and a C-terminal topoisomerase I domain, which interact to couple the hydrolysis of ATP to the overwinding of DNA. The part of the ATPase domain termed the "latch" represses topoisomerase activity in the absence of nucleotide. Here I provide evidence that the latch, in addition to its regulatory role, participates in the supercoiling mechanism during the DNA cleavage and religation steps. The latch also contributes to the coordination of ATP hydrolysis and positive supercoiling by inhibiting ATPase activity in the absence of supercoiling. The latch therefore plays an important role in the communication between the two domains of reverse gyrase. PMID- 12755602 TI - Interaction of cytochrome c with cytochrome c oxidase: an NMR study on two soluble fragments derived from Paracoccus denitrificans. AB - The functional interactions between the various components of the respiratory chain are relatively short-lived, thus allowing high turnover numbers but at the same time complicating the structural analysis of the complexes. Chemical shift mapping by NMR spectroscopy is a useful tool to investigate such transient contacts, since it can monitor changes in the electron-shielding properties of a protein as the result of temporary contacts with a reaction partner. In this study, we investigated the molecular interaction between two components of the electron-transfer chain from Paracoccus denitrificans: the engineered, water soluble fragment of cytochrome c(552) and the Cu(A) domain from the cytochrome c oxidase. Comparison of [(15)N,(1)H]-TROSY spectra of the [(15)N]-labeled cytochrome c(552) fragment in the absence and in the presence of the Cu(A) fragment showed chemical shift changes for the backbone amide groups of several, mostly uncharged residues located around the exposed heme edge in cytochrome c(552). The detected contact areas on the cytochrome c(552) surface were comparable under both fully reduced and fully oxidized conditions, suggesting that the respective chemical shift changes represent biologically relevant protein-protein interactions. PMID- 12755603 TI - Violet bioluminescence and fast kinetics from W92F obelin: structure-based proposals for the bioluminescence triggering and the identification of the emitting species. AB - Obelin from the hydroid Obelia longissima and aequorin are members of a subfamily of Ca(2+)-regulated photoproteins that is a part of the larger EF-hand calcium binding protein family. On the addition of Ca(2+), obelin generates a blue bioluminescence emission (lambda(max) = 485 nm) as the result of the oxidative decarboxylation of the bound substrate, coelenterazine. The W92F obelin mutant is noteworthy because of the unusually high speed with which it responds to sudden changes of [Ca(2+)] and because it emits violet light rather than blue due to a prominent band with lambda(max) = 405 nm. Increase of pH in the range from 5.5 to 8.5 and using D(2)O both diminish the contribution of the 405 nm band, indicating that excited state proton transfer is involved. Fluorescence model studies have suggested the origin of the 485 nm emission as the excited state of an anion of coelenteramide, the bioluminescence reaction product, and 405 nm from the excited neutral state. Assuming that the dimensions of the substrate binding cavity do not change during the excited state formation, a His22 residue within hydrogen bonding distance to the 6-(p-hydroxy)-phenyl group of the excited coelenteramide is a likely candidate for accepting the phenol proton to produce an ion-pair excited state, in support of recent suggestions for the bioluminescence emitting state. The proton transfer could be impeded by removal of the Trp92 H-bond, resulting in strong enhancement of a 405 nm band giving the violet color of bioluminescence. Comparative analysis of 3D structures of the wild-type (WT) and W92F obelins reveals that there are structural displacements of certain key Ca(2+)-ligating residues in the loops of the two C-terminal EF hands as well as clear differences in hydrogen bond networks in W92F. For instance, the hydrogen bond between the side-chain oxygen atom of Asp169 and the main-chain nitrogen of Arg112 binds together the incoming alpha-helix of loop III and the exiting alpha helix of loop IV in WT, providing probably concerted changes in these EF hands on calcium binding. But this linkage is not found in W92F obelin. These differences apparently do not change the overall affinity to calcium of W92F obelin but may account for the kinetic differences between the WT and mutant obelins. From analysis of the hydrogen bond network in the coelenterazine binding cavity, it is proposed that the trigger for bioluminescence reaction in these Ca(2+)-regulated photoproteins may be a shift of the hydrogen bond donor-acceptor separations around the coelenterazine-2-hydroperoxy substrate, initiated by small spatial adjustment of the exiting alpha-helix of loop IV. PMID- 12755604 TI - Molecular basis of spectral tuning in the newt short wavelength sensitive visual pigment. AB - Previously we reported the sequence of the member of the short wavelength sensitive 2 (SWS2) family of vertebrate visual pigments from the retina of the Japanese common newt, Cynops pyrrhogaster[Takahashi, Y. et al. (2001) FEBS Lett. 501, 151-155]. Now we have expressed the apopigment and regenerated it with A1 retinal. Its absorption maximum, 474 nm, is greatly red shifted compared to other known SWS2 pigments (418-455 nm). To determine the amino acid residues that control its spectral tuning, we replaced the residues that were near the chromophore and which differed between the newt and the bullfrog (lambda(max) = 430 nm) wild-type SWS2 pigments: Pro91Ser, Ser94Ala, Ile122Met, Cys127Ser, Ser211Cys, Tyr261Phe, and Ala292Ser. Each of these site-directed mutants led to blue shifts of the newt pigment with five of them causing substantial shifts; their sum was about equal to the difference between the absorption maximum of the bullfrog and newt pigments, 44 nm. The 32 nm shift of the absorption maximum of the multiple seven-residue mutant to 442 nm is fairly close to that of the wild type bullfrog pigment. Thus, the seven amino acid residues that we replaced are the major cause of the red shift of the newt SWS2 pigment's spectrum. Two of the residues, 91 and 94, have not previously been identified as wavelength regulating sites in visual pigments. One of these, 91, probably regulates color via a new mechanism: altering of a hydrogen bonding network that is connected via a water to the chromophore, in this case its counterion, Glu113. PMID- 12755605 TI - Analysis of flash-induced FTIR difference spectra of the S-state cycle in the photosynthetic water-oxidizing complex by uniform 15N and 13C isotope labeling. AB - Protein bands in flash-induced Fourier transform infrared (FTIR) difference spectra of the S-state cycle of photosynthetic water oxidation were analyzed by uniform (15)N and (13)C isotopic labeling of photosystem II (PS II). The difference spectra upon first- to fourth-flash illumination were obtained with hydrated (for the 1800-1200 cm(-)(1) region) or deuterated (for the 3500-3100 cm( )(1) region) films of unlabeled, (15)N-labeled, and (13)C-labeled PS II core complexes from Thermosynechococcus elongatus. Shifts of band frequencies upon (15)N and (13)C labeling provided the assignments of major peaks in the regions of 3450-3250 and 1700-1630 cm(-)(1) to the NH stretches and amide I modes of polypeptide backbones, respectively, and the assignments of some of the peaks in the 1600-1500 cm(-)(1) region to the amide II modes of backbones. Other prominent peaks in the latter region and most of the peaks in the 1450-1300 cm(-)(1) region exhibited large downshifts upon (13)C labeling but were unchanged by (15)N labeling, and hence assigned to the asymmetric and symmetric COO(-) stretching vibrations, respectively, of carboxylate groups in Glu, Asp, or the C-terminus. Peak positions corresponded well with each other among the first- to fourth-flash spectra, and most of the bands in the first- and/or second-flash spectra appeared with opposite signs of intensity in the third- and/or fourth-flash spectra. This observation indicates that the protein movements in the S(1)-->S(2) and/or S(2)- >S(3) transitions are mostly reversed in the S(3)-->S(0) and/or S(0)-->S(1) transitions, representing a catalytic role of the protein moieties of the water oxidizing complex. Drastic structural changes in carboxylate groups over the S state cycle suggest that the Asp and/or Glu side chains play important roles in the reaction mechanism of photosynthetic water oxidation. PMID- 12755606 TI - Rational design, synthesis, evaluation, and crystal structure of a potent inhibitor of human GAR Tfase: 10-(trifluoroacetyl)-5,10-dideazaacyclic-5,6,7,8 tetrahydrofolic acid. AB - Glycinamide ribonucleotide transformylase (GAR Tfase) has been the target of anti neoplastic intervention for almost two decades. Here, we use a structure-based approach to design a novel folate analogue, 10-(trifluoroacetyl)-5,10 dideazaacyclic-5,6,7,8-tetrahydrofolic acid (10-CF(3)CO-DDACTHF, 1), which specifically inhibits recombinant human GAR Tfase (K(i) = 15 nM), but is inactive (K(i) > 100 microM) against other folate-dependent enzymes that have been examined. Moreover, compound 1 is a potent inhibitor of tumor cell proliferation (IC(50) = 16 nM, CCRF-CEM), which represents a 10-fold improvement over Lometrexol, a GAR Tfase inhibitor that has been in clinical trials. Thus, this folate analogue 1 is among the most potent and selective inhibitors known toward GAR Tfase. Contributing to its efficacious activity, compound 1 is effectively transported into the cell by the reduced folate carrier and intracellularly sequestered by polyglutamation. The crystal structure of human GAR Tfase with folate analogue 1 at 1.98 A resolution represents the first structure of any GAR Tfase to be determined with a cofactor or cofactor analogue without the presence of substrate. The folate-binding loop of residues 141-146, which is highly flexible in both Escherichia coli and unliganded human GAR Tfase structures, becomes highly ordered upon binding 1 in the folate-binding site. Computational docking of the natural cofactor into this and other apo or complexed structures provides a rational basis for modeling how the natural cofactor 10 formyltetrahydrofolic acid interacts with GAR Tfase, and suggests that this folate analogue-bound conformation represents the best template to date for inhibitor design. PMID- 12755607 TI - Over-the-barrier transition state analogues and crystal structure with Mycobacterium tuberculosis purine nucleoside phosphorylase. AB - Stable chemical analogues of enzymatic transition states are imperfect mimics since they lack the partial bond character of the transition state. We synthesized structural variants of the Immucillins as transition state analogues for purine nucleoside phosphorylase and characterized them with the enzyme from Mycobacterium tuberculosis (MtPNP). PNPs form transition states with ribooxacarbenium ion character and catalyze nucleophilic displacement reactions by migration of the cationic ribooxacarbenium carbon between the enzymatically immobilized purine and phosphate nucleophiles. As bond-breaking progresses, carbocation character builds on the ribosyl group, the distance between the purine and the carbocation increases, and the distance between carbocation and phosphate anion decreases. Transition state analogues were produced with carbocation character and increased distance between the ribooxacarbenium ion and the purine mimics by incorporating a methylene bridge between these groups. Immucillin-H (ImmH), DADMe-ImmH, and DADMe-ImmG mimic the transition state of MtPNP and are slow-onset, tight-binding inhibitors of MtPNP with equilibrium dissociation constants of 650, 42, and 24 pM. Crystal structures of MtPNP complexes with ImmH and DADMe-ImmH reveal an ion-pair between the inhibitor cation and the nucleophilic phosphoryl anion. The stronger ion-pair (2.7 A) is found with DADMe-ImmH. The position of bound ImmH resembles the substrate side of the transition state barrier, and DADMe-ImmH more closely resembles the product side of the barrier. The ability to probe both substrate and product sides of the transition state barrier provides expanded opportunities to explore transition state analogue design in N-ribosyltransferases. This approach has resulted in the highest affinity transition state analogues known for MtPNP. PMID- 12755608 TI - Thermodynamic aspects and biological profile of CDAN/DOPE and DC-Chol/DOPE lipoplexes. AB - The DNA complexation and condensation properties of two established cationic liposome formulations, CDAN/DOPE (50:50, m/m; Trojene) and DC-Chol/DOPE (60:40, m/m), were investigated by using a combination of isothermal titration calorimetry (ITC), circular dichroism (CD), photon correlation spectroscopy (PCS), and turbidity assays. Plasmid DNA (7528 bp) was titrated with extruded liposomes (90 +/- 15 nm) and a thermodynamic profile established. ITC data revealed that the two liposome formulations differ substantially in their DNA complexation characteristics. Equilibrium dissociation constants for CDAN/DOPE (K(d) = 19 +/- 3 microM) and DC-Chol/DOPE liposomes (K(d) = 2 +/- 0.5 microM) were obtained by fitting the experimental data in a one-site binding model. Both CDAN/DOPE and DC-Chol/DOPE binding events take place with a negative binding enthalpy (DeltaH degrees = -0.5 and -1.7 kcal/mol, respectively) and increasing system entropy (TDeltaS = 6 +/- 0.3 and 6.2 +/- 0.3 kcal/mol, respectively). Interestingly, CDAN/DOPE liposomes undergo substantial rehydration and protonation prior to complexation with pDNA, which is observed as two discrete exothermic signals during titration. No such biphasic effects are seen with respect to the binding between DC-Chol/DOPE and pDNA that appears to be otherwise instantaneous with no rehydration effects. The rehydration and protonation characteristics of CDAN/DOPE liposomes in comparison with those of DC-Chol/DOPE cationic liposomes are confirmed by ITC; CDAN/DOPE liposomes have strongly exothermic dilution characteristics and DC-Chol/DOPE liposomes only mildly endothermic characteristics. Furthermore, analysis of cationic liposome-pDNA binding by CD spectroscopy reveals that CDAN/DOPE-pDNA lipoplexes are more structurally fluid than DC-Chol/DOPE-pDNA lipoplexes. CDAN/DOPE liposomes induced considerable fluctuation in the DNA structure for at least 60 min, whereas liposomes obtained from DC-Chol/DOPE lack the same effect on the DNA structure. Turbidity studies show that DC-Chol/DOPE lipoplexes exhibit greater resistance to serum than CDAN/DOPE lipoplexes, which showed substantial precipitation after incubation for 100 min with serum. Transfection studies on HeLa and Panc-1 cells reveal that CDAN/DOPE lipoplexes are superior in efficacy to DC-Chol/DOPE lipoplexes. CDAN/DOPE liposomes tend to transfect best in normal growth medium (including 10% serum and antibiotics), whereas DC-Chol/DOPE lipoplexes transfect best under serum free transfection conditions. PMID- 12755609 TI - Structural studies on the hairpins at the 3' untranslated region of an anthrax toxin gene. AB - Three proteins, namely, protective antigen (PA), edema factor (EF), and lethal factor (LF), encoded by the pX01 plasmid of Bacillus anthracis play a major role in the pathogenesis of target host cells. PA combines with EF and LF to form bipartite PA-EF and PA-LF toxins and facilitates intracellular delivery of EF and LF both of which cause cytotoxicity to the host. Since the level of PA is crucial to pathogenesis by anthrax toxins, it is important to understand how the host environment regulates the expression of the PA (or pagA) gene by utilizing the 5' and 3' untranslated regions (UTR). The 5' UTR sequence determines the initiation of transcription, whereas the 3' UTR sequence determines the efficient termination and stability of the transcript. Although, the role of the 5'UTR sequence of pagA has been investigated, little is known about the role of the 3' UTR. Since hairpin formation at the 3'UTR of a gene is an established mechanism for efficient termination and stability of the transcript, we carried out structural studies, including gel electrophoresis, circular dichroism, and two dimensional nuclear magnetic resonance spectroscopy, to determine whether the 3' UTR sequences of pagA also form hairpin structures. Our results unequivocally demonstrate that both the coding and the noncoding 3' UTR sequences form stable hairpin structures. It is quite likely that the hairpins at the 3'UTR may contribute to efficient termination and stability of the pagA transcript. PMID- 12755610 TI - Purification and characterization of a transmembrane domain-deleted form of lecithin retinol acyltransferase. AB - Lecithin retinol acyltransferase (LRAT) catalyzes the esterification of all-trans retinol into all-trans-retinyl ester, an essential reaction in the vertebrate visual cycle. Since all-trans-retinyl esters are the substrates for the isomerization reaction that generates 11-cis-retinoids, this esterification reaction is essential in the operation of the visual cycle. In addition, LRAT is the founder member of a series of proteins, which are of novel sequence and have unknown functions. Native LRAT is an integral membrane protein and has never been purified. To obtain a pure LRAT, the N- and C-transmembrane termini were deleted and replaced with a poly His tag for the purpose of purification. This truncated form of LRAT, referred to as tLRAT, has been expressed in bacteria and fully purified. tLRAT is catalytically active and processes all-trans-retinol at least 10-fold more efficiently than 11-cis-retinol, the precursor to the visual chromophore. While tLRAT can be robustly expressed in bacteria, it requires detergent for extraction, as the enzyme still contains hydrophobic domains, which may interact. Indeed, tLRAT can oligomerize and forms dimers. Native LRAT also forms functional homodimers. These studies pave the way for the preparation of large-scale amounts of pure tLRAT for further mechanistic and structural studies. PMID- 12755611 TI - Structure of the substrate binding pocket of the multidrug transporter EmrE: site directed spin labeling of transmembrane segment 1. AB - Site-directed spin labeling (SDSL) was used to explore the structural framework responsible for the obligatory drug-proton exchange in the Escherichia coli multidrug transporter, EmrE. For this purpose, a nitroxide scan was carried out along a stretch of 26 residues that include transmembrane segment 1 (TMS1). This segment has been implicated in the catalytic mechanism of EmrE due to the presence of the highly conserved glutamate 14, a residue absolutely required for ligand binding. Sequence-specific variation in the accessibilities of the introduced nitroxides to molecular oxygen reveals a transmembrane helical conformation along TMS1. One face of the helix is in contact with the hydrocarbon interior of the detergent micelle while the other face appears to be solvated by an aqueous environment, resulting in significant exposure of the nitroxides along this face to NiEDDA. TMS1 from two different subunits are in close proximity near a 2-fold axis of symmetry as revealed by the analysis of spin-spin interactions at sites 14 and 18. The limited extent of spin-spin interactions is consistent with a scissor-like packing of the two TMS1. This results in a V-shaped chamber which is in contact with the aqueous phase near the N-terminus. The spatial organization of TMS1, particularly the close proximity of E14, is consistent with a proposed mechanistic model of EmrE [Yerushalmi, H., and Schuldiner, S. (2000) Biochemistry 39, 14711-14719] where substrate extrusion is coupled to proton influx through electrostatic interactions and shifts of the glutamate 14 pK(a) during the cycle. PMID- 12755612 TI - Reversing the action of noncompetitive inhibitors (MK-801 and cocaine) on a protein (nicotinic acetylcholine receptor)-mediated reaction. AB - The nicotinic acetylcholine receptor (nAChR) is one of five structurally related membrane proteins required for communication between approximately 10(12) cells of the mammalian nervous system. The receptor is inhibited by both therapeutic agents and abused drugs. Understanding the mechanism of noncompetitive allosteric inhibitors of the nicotinic acetylcholine receptor is a long-standing and intensely investigated problem. During the past two decades, many attempts have been made to find drugs that prevent cocaine inhibition, including the synthesis of hundreds of cocaine analogues and derivatives, so far without success. The use of newly developed transient kinetic techniques in investigations of the inhibition of the receptor by the anticonvulsant MK-801 [(+)-dizocilpine] and the abused drug cocaine led to an inhibition mechanism not previously proposed. This mechanism indicates the properties of compounds that would prevent allosteric inhibition of the receptor and how to test for such compounds. Here we present the first evidence that small organic compounds (cocaine derivatives) exist that prevent cocaine and MK-801 inhibition of this receptor. These compounds are RTI 4229-70, a previously synthesized cocaine derivative, and based on its structure four newly synthesized cocaine derivatives, RCS-III-143, RCS-III-140A, RCS-III 218, and RCS-III-202A. Because the nAChR desensitizes rapidly, to make the required measurements a cell-flow technique with a time resolution of 10 ms was used to equilibrate BCH(3) cells containing the fetal mouse muscle-type nAChR with carbamoylcholine. The resulting whole-cell current pertaining to the nondesensitized nAChR was determined. Inhibitors and compounds that alleviate inhibition were tested by their effect on the whole-cell current. PMID- 12755613 TI - Binding of phlorizin to the isolated C-terminal extramembranous loop of the Na+/glucose cotransporter assessed by intrinsic tryptophan fluorescence. AB - Phlorizin, a phloretin 2'-glucoside, is a potent inhibitor of the Na(+)/glucose cotransporter (SGLT1). On the basis of transport studies in intact cells, a binding site for phlorizin was suggested in the region between amino acids 604 610 of the C-terminal loop 13. To further investigate phlorizin binding titration experiments of the intrinsic Trp fluorescence of isolated wild-type loop 13 and two mutated loops (Y604K and G609K) were carried out. Phlorizin (135 microM) produced approximately 40% quenching of the fluorescence of wild-type loop 13; quenching could also be observed with the two mutated loops. The apparent K(d) was lowest for the wild-type loop 13 (K(d) approximately 23 microM), followed by mutant G609K (57 microM) and mutant Y604K (70 microM). Binding of phlorizin was further confirmed by a decrease of the accessibility of loop 13 to the collisional quencher acrylamide. The interaction involves the aromatic moiety of the aglucone since phloretin (the aglucone of phlorizin) showed almost the same effects as phlorizin, while d-glucose did not. MALDI-TOF experiments revealed that loop 13 contained a disulfide bond between Cys 560 and Cys 608 that is very important for phlorizin-dependent fluorescence quenching. These studies provide direct evidence that loop 13 is a site (important amino acids including 604-609) for the molecular interaction between SGLT1 and phlorizin. They confirm that the aglucone part of the glucoside is responsible for this interaction. PMID- 12755614 TI - Growth factor-binding sequence in human alpha2-macroglobulin targets the receptor binding site in transforming growth factor-beta. AB - alpha(2)-Macroglobulin (alpha(2)M) binds transforming growth factor-beta1 (TGF beta1) and TGF-beta2, forcing these growth factors into a state of latency. The mechanism by which this occurs remains unclear. In this paper, we demonstrate that peptides, derived from the structure of human alpha(2)M (amino acids 714 729), bind directly to TGF-beta1 and block the binding of TGF-beta1 to the type I and II TGF-beta receptors. The alpha(2)M-derived peptides are notable for hydrophobic tripeptide sequences (WIW or VVV) and acidic residues (Glu(714) and Asp(719) in the mature alpha(2)M subunit), which may function analogously to the structural elements that mediate TGF-beta-binding in the type II receptor. Mutating Glu(714) and Asp(719) in the alpha(2)M-peptide-GST fusion protein, FP3, which contains the putative growth factor-binding site, significantly decreased the binding affinity of FP3 for TGF-beta1. The alpha(2)M-derived peptides, which bind TGF-beta1, inhibited the interaction of TGF-beta1 with its receptors in fetal bovine heart endothelial cells. The same peptides also inhibited the activity of TGF-beta1 in endothelial cell proliferation assays. These results demonstrate that alpha(2)M-derived peptides target the receptor-binding sequence in TGF-beta. PMID- 12755615 TI - Mechanism of inhibition of skeletal muscle actomyosin by N-benzyl-p toluenesulfonamide. AB - N-Benzyl-p-toluenesulfonamide (BTS) is a small organic molecule that specifically inhibits the contraction of fast skeletal muscle fibers. To determine the mechanism of inhibition by BTS, we performed a kinetic analysis of its effects on the elementary steps of the actomyosin subfragment-1 ATPase cycle. BTS decreases the steady-state acto-S1 ATPase rate approximately 10-fold and increases the actin concentration for half-maximal activation. BTS primarily affects three of the elementary steps of the reaction pathway. It decreases the rate of P(i) release >20-fold in the absence of actin and >100-fold in the presence of actin. It decreases the rate of S1.ADP dissociation from 3.9 to 0.8 s(-)(1) while decreasing the S1.ADP dissociation constant from 2.3 to 0.8 microM. BTS weakens the apparent affinity of S1.ADP for actin, increasing the K(d) from 7.0 to 29.5 microM. ATP binding to S1, hydrolysis, and the affinity of nucleotide-free S1 for actin are unaffected by BTS. Kinetic modeling indicates that the binding of BTS to myosin depends on actin association/dissociation and on nucleotide state. Our results suggest that the reduction of the acto-S1 ATPase rate is due to the inhibition of P(i) release, and the suppression of tension is due to inhibition of P(i) release in conjunction with the decreased apparent affinity of S1.ADP.P(i) and S1.ADP for actin. PMID- 12755616 TI - Influence of ionic strength, actin state, and caldesmon construct size on the number of actin monomers in a caldesmon binding site. AB - There is no consensus on the mechanism of inhibition of actin-myosin ATPase activity by caldesmon. Various models are based on different assumptions for the number of actin monomers that constitute a caldesmon binding site. Differences in binding behavior may be due to variations in the assay, the range of caldesmon concentrations, the type of caldesmon, and the method of data analysis used. We have evaluated these factors by measuring binding in the presence and absence of tropomyosin with both intact caldesmon and a recombinant 35 kDa actin binding fragment and with actin initially in the polymerized state or monomeric state. In all cases caldesmon binding could be simulated with a model having one class of binding sites. However, the number of actin monomers constituting a site was variable. Binding to F-actin at 165 mM ionic strength was best described with 7 actin monomers per site. When caldesmon bound to actin during the polymerization of G-actin, the size of the binding site was 3. Binding of the expressed truncated fragment, Cad35, could be described with 3 monomers per site. A simple interpretation of the data is that caldesmon binds tightly to 2-3 actin monomers. Additional parts of caldesmon bind less tightly to actin, causing caldesmon to cover approximately 7 actin monomers. The appendix contains an analysis of several binding curves with multiple binding site models. There is no compelling evidence for two classes of binding sites. PMID- 12755617 TI - Resonance Raman investigations of cytochrome c conformational change upon interaction with the membranes of intact and Ca2+-exposed mitochondria. AB - The conformational states of cytochrome c inside intact and Ca(2+)-exposed mitochondria have been investigated using resonance Raman spectroscopy. Intact and swelling bovine heart and rat liver mitochondria were examined with an excitation wavelength (413.1 nm) in resonance with the Soret transition of ferrous cytochrome c. The different b- to c-type cytochrome concentration ratio in mitochondria from two different tissues was used to help assign the Raman spectral components. Resonance Raman spectra were also recorded for mitochondria fractions (supernatants and pellets) obtained from swollen (Ca(2+)-exposed) mitochondria after differential centrifugation. The results illustrate that cytochrome c has an altered vibrational spectrum in solution, in intact, and in swollen mitochondria. When cytochrome c is released from mitochondria, its Raman spectrum becomes identical to that of ferrous cytochrome c in solution. The spectra of mitochondrial pellets indicate that a small amount of structurally modified cytochrome c remains associated with the heavy membrane fraction. Indeed, spectroscopic shifts in the low-frequency fingerprint and the high frequency marker-band regions suggest that membrane binding leads to a partial opening of the heme pocket and an alteration of the heme thioether bonds. The results support the conclusion that most cytochrome c molecules in mitochondria are membrane-bound and that the cytochrome c structure changes upon binding. Furthermore, changes in the resonance Raman active mode located at 675 cm(-)(1) in the spectra of intact, swollen, and fractionated mitochondria indicate that b type cytochromes may also undergo structural alterations during mitochondrial swelling and disruption. PMID- 12755618 TI - A cleavable affinity biotinylating agent reveals a retinoid binding role for RPE65. AB - Retinal pigment epithelial (RPE) membranes contain the full biochemical apparatus capable of processing all-trans-retinol (vitamin A) into 11-cis-retinal, the visual chromophore. As many of these proteins are integral membrane proteins and resistant to traditional methods of identification, alternate methods of identifying these proteins are sought. The approach described here involves affinity biotinylation with alkali cleavable linkers. A vitamin A containing affinity-labeling haloacetate is described which facilitates the identification of retinoid binding proteins (RBPs). Treatment of crude bovine RPE membranes with (3R)-3-[boc-lys(biotinyl)-O]-all-trans-retinol chloroacetate 1 in the low micromolar range led to the specific labeling of RPE65 and lecithin retinol acyltransferase (LRAT). Only RPE65 is labeled at 5 microM 1 at 4 degrees C. Labeled RPE65 was readily isolated by binding the labeled protein to avidin containing beads, followed by cleavage of the protein from the beads at pH 11. Trypsin digestion of RPE65 modified by 1, followed by mass spectrometry, demonstrates that C231 and C448 are alkylated by 1. These studies validate the approach that was used, and furthermore demonstrate that RPE65, a major membrane associated protein of the RPE, is a RBP. PMID- 12755620 TI - FTIR spectroscopic characterization of the cytochrome aa3 from Acidianus ambivalens: evidence for the involvement of acidic residues in redox coupled proton translocation. AB - The aa(3)-type quinol oxidase from Acidianus ambivalens is a divergent member of the heme-copper oxidases superfamily, namely, concerning the putative channels for intraprotein proton conduction. In this study, we used electrochemically induced FTIR difference spectroscopy to identify residues involved in redox coupled protonation changes. In the spectral region characteristic for the nu(C=O) mode from protonated aspartic or glutamic acid side chains, a number of prominent features can be observed between 1790 and 1710 cm(-)(1), clearly indicating the reorganization or protonation of more than four protonatable residues upon electron transfer. A direct comparison of the Fourier-transform infrared difference spectra at different pH values reveals the noteworthy high pK of >8 for some of these residues, and the protonation of two of them. These acidic residues may play a role in the proton transport to the oxygen reducing site, in proton pumping pathways, or in protonation reactions concomitant with quinone reduction. Whereas the residues contributing between 1790 and 1750 cm( )(1) have the typical position of an aspartic/glutamic acid side chain buried in the protein, a position closer to the surface is suggested for the residues contributing below approximately 1730 cm(-)(1). The possible involvement of residues contributing between 1750 and 1720 cm(-)(1) in the quinone binding site is demonstrated on the basis of experiments in the presence and absence of ubiquinone-2 and of the native electron carrier of the A. ambivalens respiratory chain, caldariella quinone. Most signals seen here are not observable in comparable spectra of typical members of the heme copper oxidase superfamily and thus reflect unique features of the enzyme from the hyperthermoacidophilic archaeon A. ambivalens. PMID- 12755619 TI - Structural study of GCDFP-15/gp17 in disease versus physiological conditions using a proteomic approach. AB - Gross cystic disease fluid protein (GCDFP-15), also known as prolactin-inducible protein (PIP), is a specific breast tumor marker. GCDFP-15/PIP is also identified as gp17 and/or seminal actin-binding protein (SABP) from seminal vesicles and as extraparotid glycoprotein (EP-GP) from salivary glands. It is an aspartyl proteinase able to specifically cleave fibronectin (FN), suggesting a possible involvement in mammary tumor progression and fertilization. Other functions were attributed to this protein(s) on the basis of its ability to interact with an array of molecules such as CD4, actin, and fibrinogen. We investigated the structure of the protein(s) under disease versus physiological conditions by RP HPLC chromatography, ProteinChip technology, and QStar MS/MS mass spectrometry. The proteins behaved differently when examined by RP-HPLC chromatography and surface-enhanced laser desorption ionization time-of-flight (SELDI-TOF) mass spectrometry, suggesting different conformations and/or tissue-specific posttranslational modifications of the proteins, although their primary structure was identical by MS/MS analysis. Both showed a single N-glycosylation site. A different N-linked glycosylation pattern was observed in pathological GCDFP 15/PIP as compared with physiological gp17/SABP protein by coupling enzymatic digestion and ProteinChip technology. Furthermore, taking advantage of ProteinChip technology, we analyzed the interaction of both proteins with CD4 and FN. We observed that the physiological form was mainly involved in the binding to CD4. Moreover, we defined the specific FN binding-domain of this protein. These data suggested that, depending on its conformational state, the protein could differently bind to its various binding molecules and change its function(s) in the microenviroments where it is expressed. PMID- 12755621 TI - pH dependence of the donor side reactions in Ca2+-depleted photosystem II. AB - We have studied how low pH affects the water-oxidizing complex in Photosystem II when depleted of the essential Ca(2+) ion cofactor. For these samples, it was found that the EPR signal from the Y(Z)(*) radical decays faster at low pH than at high pH. At 20 degrees C, Y(Z)(*) decays with biphasic kinetics. At pH 6.5, the fast phase encompasses about 65% of the amplitude and has a lifetime of approximately 0.8 s, while the slow phase has a lifetime of approximately 22 s. At pH 3.9, the kinetics become totally dominated by the fast phase, with more than 90% of the signal intensity operating with a lifetime of approximately 0.3 s. The kinetic changes occurred with an approximate pK(a) of 4.5. Low pH also affected the induction of the so-called split radical EPR signal from the S(2)Y(Z)(*) state that is induced in Ca(2+)-depleted PSII membranes because of an inability of Y(Z)(*) to oxidize the S(2) state. At pH 4.5, about 50% of the split signal was induced, as compared to the amplitude of the signal that was induced at pH 6.5-7, using similar illumination conditions. Thus, the split-signal induction decreased with an apparent pK(a) of 4.5. In the same samples, the stable multiline signal from the S(2) state, which is modified by the removal of Ca(2+), was decreased by the illumination to the same extent at all pHs. It is proposed that decreased induction of the S(2)Y(Z)(*) state at lower pH was not due to inability to oxidize the modified S(2) state induced by the Ca(2+) depletion. Instead, we propose that the low pH makes Y(Z)(*) able to oxidize the S(2) state, making the S(2) --> S(3) transition available in Ca(2+)-depleted PSII. Implications of these results for the catalytic role of Ca(2+) and the role of proton transfer between the Mn cluster and Y(Z) during oxygen evolution is discussed. PMID- 12755622 TI - Binding of manganese stabilizing protein to photosystem II: identification of essential N-terminal threonine residues and domains that prevent nonspecific binding. AB - The N-terminus of spinach photosystem II manganese stabilizing protein (MSP) contains two amino acid sequences, (4)KRLTYD(10)E and (15)TYL(18)E, that are necessary for binding of two copies of this subunit to the enzyme [Popelkova et al. (2002) Biochemistry 41, 10038-10045]. To better understand the basis of MSP photosystem II interactions, the role of threonine residues in the highly conserved motifs T(Y/F)DE and TY has been characterized. Deletion mutants lacking the first 5, 6, 7, and 15 amino acid residues at the N-terminus of the protein were examined for their ability to reconstitute activity in MSP-depleted photosystem II. The results reported here show that truncations of five and six amino acid residues (mutants DeltaR5M and DeltaL6M mutants) have no negative effect on recovery of oxygen evolution activity or on binding of MSP to photosystem II. Deletion of seven residues (mutant DeltaT7M) decreases reconstitution activity to 40% of the control value and reduces functional binding of the mutant protein to photosystem II from two to one copy. Deletion of 15 amino acid residues (mutant DeltaT15M) severely impairs functional binding of MSP, and lowers O(2) evolution activity to less than 20% of the control. DeltaT7M is the only mutant that exhibited neither nonspecific binding to photosystem II nor changes in tertiary structure. These, and previous results, show that the highly conserved Thr7 and Thr15 residues of MSP are required for functional binding of two copies of the eukaryotic protein to photosystem II. Although the N terminal domains, (1)EGGKR(6)L, (8)YDEIQS(14)K, and (16)YL(18)E of spinach MSP are unnecessary for specific, functional binding interactions, these sequences are necessary to prevent nonspecific binding of the protein to photosystem II. PMID- 12755623 TI - EPR and ENDOR evidence for a 1-His, hydroxo-bridged mixed-valent diiron site in Desulfovibrio vulgaris rubrerythrin. AB - Key features differentiating the coordination environment of the two irons in the mixed-valent (Fe(2+),Fe(3+)) diiron site of Desulfovibrio vulgaris rubrerythrin (Rbr(mv)) were determined by continuous wave (CW) and pulsed ENDOR spectroscopy at 35GHz. (14)N ENDOR evidence indicates that a nitrogen is bound only to the Fe(2+) ion of the mixed-valent site. Assuming that this nitrogen is from His131Ndelta, the same one that furnishes an iron ligand in the crystal structure of the diferric site, the ENDOR data allow us to specify the Fe(2+) and Fe(3+) positions within the molecular reference frame. In addition, the (1,2)H ENDOR on Rbr(mv) indicates the presence of a solvent-derived aqua/hydroxo ligand bound either terminally or in a bridging mode to Fe(3+) in the mixed-valent site. The relatively large g anisotropy of Rbr(mv) and weak antiferromagnetic coupling, J approximately -8 cm(-)(1) (in the 2JS(1)*S(2) formalism), between the irons is more consistent with a bridging than terminal hydroxo ligand. gamma-Irradiation was used to cryoreduce Rbr at 77 K, thereby producing a mixed-valent diiron site [(Rbr(ox))(mv)] that retains the structure of the diferric site. The EPR spectrum of (Rbr(ox))(mv) was nearly identical to that of the as-isolated or chemically reduced samples. This near identity implies that the structure of the mixed valent Rbr diiron site is essentially identical to that of the diferric site, except for protonation of the oxo bridge, which apparently occurred via a proton jump from hydrogen-bonded solvent at 77 K. The EPR spectrum of (Rbr(ox))(mv) thus supports the (14)N ENDOR-assigned His131 ligation to Fe(2+) and assignment of the solvent-derived ligand observed in the (1,2)H ENDOR to a hydroxo bridge between the irons of the mixed-valent diiron site. PMID- 12755624 TI - (18)O isotope exchange measurements reveal that calcium is involved in the binding of one substrate-water molecule to the oxygen-evolving complex in photosystem II. AB - Direct evidence is presented to show that calcium is inherently involved in the binding of one of the two substrate-water molecules to the oxygen-evolving complex in photosystem II. Previous rapid (millisecond range) (18)O isotope exchange measurements between added H(2)(18)O and the photogenerated O(2) have shown that the two substrate-water molecules bind to separate sites throughout the S-state cycle, as revealed by their kinetically distinct rates of (18)O exchange [Hillier, W., and Wydrzynski, T. (2000) Biochemistry 39, 4399-4405]. Upon extraction of the functionally bound calcium using a either a low-pH/citrate treatment or a NaCl/A23187/EGTA treatment and subsequent reconstitution of activity with strontium, we show for the first time a specific increase in the slow rate of (18)O exchange by a factor of 3-4. This increase in the slow rate of exchange is consistently observed across the S(1), S(2), and S(3) states. In contrast, the fast phase of (18)O exchange in the S(3) state appears to be affected little upon strontium reconstitution, while the fast phases of exchange in the S(1) and S(2) states remain largely unresolvable, at the detectable limits of the current techniques. The results are discussed in terms of a possible substrate bridging structure between the functional calcium and a catalytic manganese ion that gives rise to the slowly exchanging component. PMID- 12755625 TI - Specific antibody-DNA interaction: a novel strategy for tight DNA recognition. AB - Anti-double-stranded DNA monoclonal antibodies against a viral transcriptional regulatory site are capable of discriminating single-base replacements with affinities of 1 x 10(-)(9) M, which were optimized for the length of the duplex used as the immunogen. Their affinity for DNA duplexes of increasing length is lower, but reaches a plateau at 2 x 10(-)(8) M, still a fairly high affinity compared to those of most known natural anti-DNA antibodies. The ability of the antibodies to bind to a 166 bp DNA fragment containing the specific sequence strongly suggests that these have the potential of binding the specific sequence within larger genomic DNA fragments. Electrostatic interactions do not play a significant role, the opposite of what is observed in natural DNA binding interfaces. In addition, the insensitivity of the antibody-DNA interaction to solute effects is indicative of a marginal participation of water molecules at the interface compared to the level of participation at the natural E2-DNA interface. Spectroscopic evidence of base unstacking strongly suggests substantial denaturation of antibody-bound DNA, in agreement with thermodynamic results that show an unusual positive heat capacity change, which could be explained at least in part by the exposure of DNA bases upon binding. Lower local DNA stability cooperates with sequence recognition in producing the highest binding affinity. A slow rate of antibody-DNA association indicates an energy barrier imposed by conformational rearrangements, as opposed to an electrostatically assisted diffusion-controlled collision in the E2 DNA binding domain. While the E2-DNA interaction takes place through a typical direct readout mechanism, the anti-double-stranded DNA monoclonal antibody-DNA interaction could be viewed as a distinctive case of indirect readout with a significant distortion in the DNA conformation. However, the precise mechanism with which the DNA bases are accommodated in the antibody combining site will require structural analysis at atomic resolution. These results constitute a first stage for unveiling the unusual molecular recognition mechanism of a specific DNA sequence by antibodies. This mechanism could represent the strategy with which the immune system tightly and specifically recognizes a DNA antigen. PMID- 12755626 TI - Amphotericin B binds to amyloid fibrils and delays their formation: a therapeutic mechanism? AB - The membrane-active antifungal agent amphotericin B (AmB) is one of the few agents shown to slow the course of prion diseases in animals. Congo Red and other small molecules have been reported to directly inhibit amyloidogenesis in both prion and Alzheimer peptide model systems via specific binding. We propose that it is possible that AmB may act similarly to physically prevent conversion of the largely alpha-helical prion protein (PrP) to the pathological beta-sheet aggregate protease-resistant isoform (PrP(res)) in prion disease and by analogy prevent fibrillization in amyloid diseases. To assess whether AmB is capable of binding specifically to amyloid fibrils as does Congo Red, we have used the insulin fibril and Abeta 25-35 amyloid model fibril system. We find that AmB does bind strongly to both insulin (K(d) = 1.1 microM) and Abeta 25-35 amyloid (K(d) = 6.4 microM) fibrils but not to native insulin. Binding is characterized by a red shifted AmB spectrum indicative of a more hydrophobic environment. Thus AmB seems to have a complementary face for amyloid fibrils but not the native protein. In addition, AmB interacts specifically with Congo Red, a known fibril-binding agent. In kinetic fibril formation studies, AmB was able to significantly kinetically delay the formation of Abeta 25-35 fibrils at pH 7.4 but not insulin fibrils at pH 2. PMID- 12755627 TI - Quantitative analysis of influenza virus RNP interaction with RNA cap structures and comparison to human cap binding protein eIF4E. AB - Influenza virus polymerase uses capped RNA primers for transcription initiation in infected cells. This unique mechanism involves the specific binding of the polymerase to capped mRNA precursors in the nucleus of infected cells. These host RNAs are then cleaved by a polymerase associated endonuclease at a position 10-15 nucleotides downstream of the cap structure. The resulting capped RNA oligonucleotides function as primers for transcription initiation. The viral cap binding site has previously been mapped to the PB2 subunit of the trimeric influenza polymerase complex. We have established a quantitative assay system for the analysis of cap interaction with PB2 as part of the native, viral ribonucleoprotein complex (RNP) using a specific UV cross-linking approach. Cap binding was not affected by the RNase pretreatment of the capped RNA substrate and cap binding was not inhibited by excess uncapped RNA, indicating that under the assay conditions, the majority of the binding energy was contributed by the interaction with the cap structure. Binding to 7-methyl-GTP was found to involve synergistic interaction with 7-methyl guanosine and triphosphate binding subsites. A similar mode of interaction with 7-methyl-GTP was found for human cap binding protein eIF4E. However, the potency of 7-methyl-GTP for cap binding inhibition was 200-fold stronger with eIF4E and had a higher contribution from the triphosphate moiety as compared to influenza RNP. Due to this difference in cap subsite interaction, it was possible to identify novel cap analogues, which selectively interact with influenza virus, but not human cap binding protein. PMID- 12755628 TI - BmSPN2, a serpin secreted by the filarial nematode Brugia malayi, does not inhibit human neutrophil proteinases but plays a noninhibitory role. AB - The filarial nematode, Brugia malayi, is a causative agent of lymphatic filariasis. Bm-spn-2, one of two serpin genes identified in B. malayi, is expressed only in humans where the encoded protein, BmSPN2, is secreted by blood dwelling microfilariae. Previous work reported that BmSPN2 could inhibit the activities of elastase and cathepsin G from human neutrophils, despite an atypical amino acid sequence. This did not fit with accepted theories as to the sequence requirements of serpins for proteinase inhibition. We have cloned and expressed Bm-spn-2 in Escherichia coli and characterized the structural and functional properties of recombinant BmSPN2. Sequence alignment, circular dichroism spectroscopy, and susceptibility to cleavage by proteinases all suggest that BmSPN2 shares the tertiary structure typical of the serpin family including an accessible reactive center loop. However, we have found that BmSPN2 has no effect on the activity of neutrophil elastase or cathepsin G and does not form SDS-stable complexes with these proteinases. We provide evidence that BmSPN2 cannot undergo the characteristic stressed to relaxed transition required for proteinase inhibition by serpins. We conclude that BmSPN2 is not an atypical inhibitor but is a new noninhibitory serpin, in keeping with its sequence. PMID- 12755629 TI - Inhibition of Moloney murine leukemia virus integration using polyamides targeting the long-terminal repeat sequences. AB - The retroviral integrase (IN) carries out the integration of the viral DNA into the host genome. Both IN and the DNA sequences at the viral long-terminal repeat (LTR) are required for the integration function. In this report, a series of minor groove binding hairpin polyamides targeting sequences within terminal inverted repeats of the Moloney murine leukemia virus (M-MuLV) LTR were synthesized, and their effects on integration were analyzed. Using cell-free in vitro integration assays, polyamides targeting the conserved CA dinucleotide with cognate sites closest to the terminal base pairs were effective at blocking 3' processing but not strand transfer. Polyamides which efficiently inhibited 3' processing and strand transfer targeted the LTR sequences through position 9. Polyamides that inhibited integration were effective at nanomolar concentrations and showed subnanomolar affinity for their cognate LTR sites. These studies highlight the role of minor groove interactions within the LTR termini for retroviral integration. PMID- 12755630 TI - Absence of evidence for metabolite-modulated association between alpha-glycerol-3 phosphate dehydrogenase and L-lactate dehydrogenase. AB - Evidence for the NADH-modulated formation of a complex between alpha-glycerol-3 phosphate dehydrogenase and l-lactate dehydrogenase was reported [Yong, H., Thomas, G. A., and Peticolas, W. L. (1993) Biochemistry 32, 11124-11131]. This NADH-modulated association suggested a mechanism of potentially great importance to enzyme modulation and the controversial phenomena of direct NADH channeling. In the present paper, we reproduce with additional controls the experiments described by Yong et al. ((1993) Biochemistry 32, 11124-11131). Our results conclusively demonstrate the absence of detectable association between alpha glycol-3-phosphate dehydrogenase and l-lactate dehydrogenase. PMID- 12755631 TI - Structural and morphological diversity of (1-->3)-beta-D-glucans synthesized in vitro by enzymes from Saprolegnia monoica. Comparison with a corresponding in vitro product from blackberry (Rubus fruticosus). AB - Detergent extracts of microsomal fractions from Saprolegnia monoica and blackberry (Rubus fruticosus) cells were incubated with UDP-glucose to yield in vitro (1-->3)-beta-d-glucans. The insoluble products were analyzed by conventional and cryo transmission electron microscopy, X-ray diffraction, and (13)C CP/MAS NMR, and their molecular weights were determined by light scattering experiments. All the products were microfibrillar, but for the detergent extracts from S. monoica, important morphological differences were observed when the pH of the synthesizing medium was modified. At pH 6, the product had a weight average degree of polymerization () exceeding 20 000 and consisted of endless ribbon-like microfibrils. The microfibrils obtained at pH 9 had a length of only 200-300 nm, and their was approximately 5000. Of all the in vitro (1-->3)-beta-d-glucans, the one from R. fruticosus had the shortest length and the smallest. Crystallographic and spectroscopic data showed that the three in vitro samples consisted of triple helices of (1-->3)-beta-d-glucans and contained substantial amounts of water molecules in their structure, the shortest microfibrils being more hydrated. In addition, the long microfibrils from S. monoica synthesized at pH 6 were more resistant toward the action of an endo-(1-->3)-beta-d-glucanase than the shorter ones obtained at pH 9. These results are discussed in terms of molecular biosynthetic mechanisms of fungal and plant (1-->3)-beta-d-glucans, and in relation with the possible existence of several (1-->3)-beta-d-glucan synthases in a given organism. The interpretation and discussion of these observations integrate the current knowledge of the structure and function of (1-->3)-beta-d glucans. PMID- 12755632 TI - Glucose-induced stimulation of the Ras-cAMP pathway in yeast leads to multiple phosphorylations and activation of 6-phosphofructo-2-kinase. AB - Yeast cells respond to changes of the environment by complex modifications of the metabolism. An increase of the extracellular glucose concentration activates the Ras-cAMP pathway. Via a production of cAMP this pathway stimulates the cAMP dependent protein kinase (PKA) which is involved in the posttranslational regulation of the key enzymes of gluconeogenesis and glycolysis. 6-Phosphofructo 2-kinase (PFK2) catalyzes the synthesis of fructose 2,6-bisphosphate, the most potent activator of the glycolytic key enzyme 6-phosphofructo-1-kinase. We investigated the molecular mechanism of the glucose-induced phosphorylation and activation of PFK2 in Saccharomyces cerevisiae. After an incubation of PFK2 with ATP and PKA in vitro, two amino acid residues, Thr157 and Ser644, are phosphorylated and the enzyme is activated. A stimulation of the Ras-cAMP pathway by glucose addition to cultivated yeast cells leads to an in vivo activation of PFK2 which is accompanied by a more complex phosphorylation pattern of the enzyme. The phosphorylation of the protein on Ser644 is the result of PKA stimulation while the protein kinase(s) catalyzing the 5-fold phosphorylation of the peptide fragment T(67)(-)(101) is (are) still unknown. The functional significance of T(67)(-)(101) and its phosphorylation is supported by the finding that PFK2 lacking this peptide is inactive. PMID- 12755633 TI - Kinetic and spectroscopic characterization of the H178A methionyl aminopeptidase from Escherichia coli. AB - To gain insight into the role of the strictly conserved histidine residue, H178, in the reaction mechanism of the methionyl aminopeptidase from Escherichia coli (EcMetAP-I), the H178A mutant enzyme was prepared. Metal-reconstituted H178A binds only one equivalent of Co(II) or Fe(II) tightly with affinities that are identical to the WT enzyme based on kinetic and isothermal titration calorimetry (ITC) data. Electronic absorption spectra of Co(II)-loaded H178A EcMetAP-I indicate that the active site divalent metal ion is pentacoordinate, identical to the WT enzyme. These data indicate that the metal binding site has not been affected by altering H178. The effect of altering H178 on activity is, in general, due to a decrease in k(cat). The k(cat) value for Co(II)-loaded H178A decreased 70-fold toward MGMM and 290-fold toward MP-p-NA compared to the WT enzyme, while k(cat) decreased 50-fold toward MGMM for the Fe(II)-loaded H178A enzyme and 140-fold toward MP-p-NA. The K(m) values for MGMM remained unaffected, while those for MP-p-NA increased approximately 2-fold for Co(II)- and Fe(II) loaded H178A. The k(cat)/K(m) values for both Co(II)- and Fe(II)-loaded H178A toward both substrates ranged from approximately 50- to 580-fold reduction. The pH dependence of log K(m), log k(cat), and log(k(cat)/K(m)) of both WT and H178A EcMetAP-I were also obtained and are identical, within error, for H178A and WT EcMetAP-I. Therefore, H178A is catalytically important but is not required for catalysis. Assignment of one of the observed pK(a) values at 8.1 for WT EcMetAP-I was obtained from plots of molar absorptivity at lambda(max(640)) vs pH for both WT and H178A EcMetAP-I. Apparent pK(a) values of 8.1 and 7.6 were obtained for WT and H178A EcMetAP-I, respectively, and were assigned to the deprotonation of a metal-bound water molecule. The data reported herein provide support for the key elements of the previously proposed mechanism and suggest that a similar mechanism can apply to the enzyme with a single metal in the active site. PMID- 12755634 TI - Interaction of monodisperse anionic amphiphiles with the i-face of secreted phospholipase A2. AB - Pancreatic IB phospholipase A(2) (PLA2) forms aggregates of defined size with monodisperse alkyl sulfates in the premicellar concentration range. As an extension of the interfacial kinetic paradigm, results are interpreted in terms of a model in which several amphiphile molecules bind along their polar headgroup to the interface binding region (i-face) of PLA2. The resulting complex, E(#), has a half-micellar structure, and it acts as an "amphiphile" in the aqueous phase. E(#) not only self-aggregates but also binds hydrophobic probes and interacts with hydrophobic surfaces. As expected, resonance energy transfer from the tryptophan donor in PLA2 to an acceptor probe partitioned in E(#) shows a biphasic dependence as the probe coexisting with PLA2 is diluted at higher alkyl sulfate concentrations. The gel-permeation behavior of PLA2 at premicellar alkyl sulfate concentrations is also biphasic. For example, above 1.2 mM decyl sulfate (CMC = 3.5 mM) PLA2 elutes as a single sharp peak, presumably the self-aggregate of E(#) with apparent molecular mass of 120-150 kDa. At 0.4-1 mM decyl sulfate the retention volume is even larger than that for the 14 kDa PLA2. This anomalous retention is attributed to the interaction of the hydrophobic region of E(#) with the hydrophobic patches on the gel-permeation matrix. Elution behavior of the self-aggregated E(#) form of site-directed mutants in dodecyl sulfate suggests that certain substitutions in the conserved hydrogen-bonding network have a significant effect on the aggregate size. These results suggest a role for the network in the amphiphile binding along the i-face of PLA2, presumably through a change in the anion coordination ligands. PMID- 12755635 TI - Thermodynamic analysis of binding between mouse major urinary protein-I and the pheromone 2-sec-butyl-4,5-dihydrothiazole. AB - The mouse pheromone 2-sec-butyl-4,5-dihydrothiazole (SBT) binds to an occluded, nonpolar cavity in the mouse major urinary protein-I (MUP-I). The thermodynamics of this interaction have been characterized using isothermal titration calorimetry (ITC). MUP-I-SBT binding is accompanied by a large favorable enthalpy change (DeltaH = -11.2 kcal/mol at 25 degrees C), an unfavorable entropy change ( TDeltaS = 2.8 kcal/mol at 25 degrees C), and a negative heat capacity change [DeltaC(p)() = -165 cal/(mol K)]. Thermodynamic analysis of binding between MUP-I and several 2-alkyl-4,5-dihydrothiazole ligands indicated that the alkyl chain contributes more favorably to the enthalpy and less favorably to the entropy of binding than would be expected on the basis of the hydrophobic desolvation of short-chain alcohols. However, solvent transfer experiments indicated that desolvation of SBT is accompanied by a net unfavorable change in enthalpy (DeltaH = +1.0 kcal/mol) and favorable change in entropy (-TDeltaS = -1.8 kcal/mol). These results are discussed in terms of the possible physical origins of the binding thermodynamics, including (1) hydrophobic desolvation of both the protein and the ligand, (2) formation of a buried water-mediated hydrogen bond network between the protein and ligand, (3) formation of strong van der Waals interactions, and (4) changes in the structure, dynamics, and/or hydration of the protein upon binding. PMID- 12755636 TI - Equilibrium and kinetic folding of an alpha-helical Greek key protein domain: caspase recruitment domain (CARD) of RICK. AB - We have characterized the equilibrium and kinetic folding of a unique protein domain, caspase recruitment domain (CARD), of the RIP-like interacting CLARP kinase (RICK) (RICK-CARD), which adopts a alpha-helical Greek key fold. At equilibrium, the folding of RICK-CARD is well described by a two-state mechanism representing the native and unfolded ensembles. The protein is marginally stable, with a DeltaG(H)()2(O) of 3.0 +/- 0.15 kcal/mol and an m-value of 1.27 +/- 0.06 kcal mol(-1) M(-1) (30 mM Tris-HCl, pH 8, 1 mM DTT, 25 degrees C). While the m value is constant, the protein stability decreases in the presence of moderate salt concentrations (below 200 mM) and then increases at higher salt concentrations. The results suggest that electrostatic interactions are stabilizing in the native protein, and the favorable Coulombic interactions are reduced at low ionic strength. Above 200 mM salt, the results are consistent with Hofmeister effects. The unfolding pathway of RICK-CARD is complex and contains at least three non-native conformations. The refolding pathway of RICK-CARD also is complex, and the data suggest that the unfolded protein folds via two intermediate conformations prior to reaching the native state. Overall, the data suggest the presence of kinetically trapped, or misfolded, species that are on pathway both in refolding and in unfolding. PMID- 12755637 TI - DNA binding mode of the cis and trans geometries of new antitumor nonclassical platinum complexes containing piperidine, piperazine, or 4-picoline ligand in cell-free media. Relations to their activity in cancer cell lines. AB - The global modification of mammalian and plasmid DNAs by novel platinum compounds, cis- or trans-[PtCl(2)(NH(3))(Am)], where Am = NH(3), nonplanar heterocycle piperidine, piperazine, or aromatic planar heterocycle 4-picoline, was investigated in cell-free media using various biochemical and biophysical methods. These modifications have been compared with the activity of these new compounds in several tumor cell lines including those resistant to antitumor cis diamminedichloroplatinum(II) (cisplatin). The results show that the replacement of the NH(3) group in cisplatin by the heterocyclic ligands does not considerably affect the DNA binding mode of this drug. Cytotoxicity studies have revealed that the replacement lowers the activity of the platinum compound in both sensitive and resistant cell lines. It has been suggested that the reduced activity of these analogues of cisplatin is associated with some features of the damaged DNA and/or its cellular processing. Alternatively, the reduced activity of the analogues of cisplatin might also be due to the factors that do not operate directly at the level of the target DNA, such as intracellular platinum uptake. In contrast to the analogues of cisplatin, the replacement of one ammine group by the heterocyclic ligand in its clinically ineffective trans isomer (transplatin) results in a radical enhancement of its activity in tumor cell lines. Importantly, this replacement also markedly alters the DNA binding mode of transplatin. The results support the view that one strategy of how to activate the trans geometry in bifunctional platinum(II) compounds including circumvention of resistance to cisplatin may consist of a chemical modification of the ineffective transplatin that results in an increased stability of its intrastrand cross-links in double-helical DNA and/or in an increased efficiency to form interstrand cross-links. PMID- 12755638 TI - Electron transmission through organized organic thin films. AB - Low-energy electron photoemission spectroscopy (LEPS) allows the study of the electronic properties of organized organic thin films (OOTF) adsorbed on conducting surfaces by monitoring the energy and angular distribution of electrons emitted from the substrate and transmitted through the film. The transmission properties are explained by electronic band structure in the organic film. This band is an example of an electron resonance that is delocalized in the layer. It results from the two-dimensional nature of the layer. Other resonances in the transmission spectra are also discussed, as well as their experimental manifestation. The temperature dependence of the electron transmission efficiency is explained in terms of dependence of the transmission probability on the initial momentum of the electron and on the relative orientation of the electron velocity and the molecules in the film. Hence, despite the fact that the molecules in the OOTF are weakly interacting, when not charged, the electron transmission through the film is governed by cooperative effects. These effects must be taken into account when considering electronic properties of adsorbed layers. PMID- 12755639 TI - Tyrosinase autoactivation and the chemistry of ortho-quinone amines. AB - Tyrosinase oxidizes tyrosine to dopaquinone, which undergoes nonenzymatic reactions leading to precursors of melanin pigments. Cyclization of dopaquinone gives cyclodopa, which participates in redox exchange with dopaquinone to give the eumelanin precursor dopachrome plus dopa. The indirect formation of the catechol (dopa) from the phenol (tyrosine) leads to unusual enzyme kinetics. Using a combination of enzyme oximetry, pulse radiolysis, and chemical oxidation, the study of structurally modified dopaquinones provides firm evidence of nonenzymatic catechol formation during tyrosinase oxidation of phenols and reveals significant differences in their modes of reaction. PMID- 12755640 TI - Assembly of cytochrome c oxidase within the mitochondrion. AB - Cytochrome c oxidase (CcO) is an oligomeric complex localized within the mitochondrial inner membrane. Assembly of the active oxidase complex requires the coordinate assembly of subunits synthesized in both the cytoplasm and the mitochondrion. In addition, assembly is dependent on the insertion of five types of cofactors, including two hemes, three copper ions, and one Zn, Mg, and Na ion. A series of accessory proteins are critical for synthesis of the heme A cofactor and insertion of the copper ions. This Account will focus on the steps in the coordinate assembly of CcO subunits, the formation of heme A, and the delivery and insertion of copper ions. PMID- 12755641 TI - The mechanism of methanol to hydrocarbon catalysis. AB - The process of converting methanol to hydrocarbons on the aluminosilicate zeolite HZSM-5 was originally developed as a route from natural gas to synthetic gasoline. Using other microporous catalysts that are selective for light olefins, methanol-to-olefin (MTO) catalysis may soon become central to the conversion of natural gas to polyolefins. The mechanism of methanol conversion proved to be an intellectually challenging problem; 25 years of fundamental study produced at least 20 distinct mechanisms, but most did not account for either the primary products or a kinetic induction period. Recent experimental and theoretical work has firmly established that methanol and dimethyl ether react on cyclic organic species contained in the cages or channels of the inorganic host. These organic reaction centers act as scaffolds for the assembly of light olefins so as to avoid the high high-energy intermediates required by all "direct" mechanisms. The rate of formation of the initial reaction centers, and hence the duration of the kinetic induction period, can be governed by impurity species. Secondary reactions of primary olefin products strongly reflect the topology and acid strength of the microporous catalyst. Reaction centers form continuously through some secondary pathways, and they age into polycyclic aromatic hydrocarbons, eventually deactivating the catalyst. It proves useful to consider each cage (or channel) with its included organic and inorganic species as a supramolecule that can react to form various species. This view allows us to identify structure activity and structure selectivity relationships and to modify the catalyst with degrees of freedom that are more reminiscent of homogeneous catalysis than heterogeneous catalysis. PMID- 12755642 TI - Microwave spectroscopy and nuclear magnetic resonance spectroscopy--what is the connection? AB - The history and development of microwave spectroscopy and nuclear magnetic resonance (NMR) spectroscopy have much in common. In this Account, we discuss the less widely appreciated connections between the parameters measured using the two techniques. Selected examples from our laboratory and from the recent literature attest to the utility and importance of these connections. For example, how are nuclear spin-rotation tensors and NMR chemical shifts related? Why should chemists be interested in absolute magnetic shielding scales? What can chemists learn about trends in spin-spin coupling constants from the hyperfine parameters measured in microwave and molecular beam experiments? The increasingly important role of quantum-chemical calculations in the interpretation of the microwave and NMR data is also highlighted. PMID- 12755643 TI - Chemical approaches to triggerable lipid vesicles for drug and gene delivery. AB - Effective drug delivery requires the precise spatial and temporal delivery of therapeutic agents to the target site. To this end, a variety of chemical and physicochemical approaches have been devised to create lipid vesicles (liposomes) that can be triggered to release their contents in a controlled fashion. The triggers include changes in pH, redox potential, temperature, or the level of specific enzymes. We review the chemistries that have recently been applied to exploit the pH and redox potential triggers so as to release vesicle contents in the appropriate biological location. PMID- 12755644 TI - The building block approach to unusual alpha-amino acid derivatives and peptides. AB - The building block approach was identified as a useful alternative to the commonly used Bucherer-Berg method for the preparation of cyclic unusual alpha amino acid derivatives. The symmetrical building blocks were prepared by dialkylation of ethyl isocyanoacetate under solid-liquid phase transfer catalysis conditions while the unsymmetrical building blocks were prepared by a stepwise alkylation of the O'Donnell Schiff base. Metathesis reactions, Suzuki couplings, and cycloaddition reactions were utilized to assemble the building blocks. PMID- 12755645 TI - Physical symptoms of depression as a public health concern. AB - Depressive disorders are a public health problem. They occur frequently, and it is highly likely that their prevalence will grow in the years to come. Depressive disorders can have severe consequences in terms of suffering, disability, and increased mortality, particularly if left untreated. They are present in all cultural settings and present a major difficulty for the normal functioning of patients' families. A large proportion of people with depressive disorders do not get treatment, and a major reason depressive disorders go unrecognized is that they often present mainly physical symptoms. The fact that depression often co occurs with physical illness further complicates the diagnosis and treatment of depressive disorders. Better undergraduate education of medical students and general education for the public in understanding and treating depressive disorders could considerably improve the prognosis of patients suffering from these illnesses. PMID- 12755646 TI - Physical symptoms of depression: unmet needs. AB - The burden of depression on society is sizable. Innate to this burden are underdiagnosis and under-treatment of unipolar and bipolar major depressive disorder in all parts of the health care system in part due to underrecognition of the physical symptoms that commonly are core components of major depressive disorder. Physical pains especially complicate the diagnosis of depression. Many patients de-emphasize psychosocial symptoms while emphasizing pains as their primary or sole complaints. There is a high correlation between the number of physical symptoms reported and the presence of depression. Additionally, patients with residual physical and emotional symptoms following treatment for depression appear to be at higher risk of relapse compared with those who have no residual symptoms. Complex genetic vulnerabilities underlie the depressive diathesis, and stress appears to be an accentuation for the gene expression that sets off episodes of depression in persons with these predispositions. If underdiagnosis interferes and acute treatment is not implemented early and effectively for initial episodes of depression and maintained after remission, individuals with genetic vulnerabilities may experience a pattern of recurrences, cycle acceleration, and increased severity. Serotonin and norepinephrine may be shared neurochemical links that tie depression and physical symptoms together. Thus, it is reasonable to hypothesize that antidepressants that incorporate both serotonin and norepinephrine reuptake inhibition might be a more efficacious treatment approach for patients with physical symptoms of depression. PMID- 12755647 TI - Physical symptoms of depression: unmet needs in special populations. AB - Over two thirds of people suffering from depression complain of pain with or without reporting psychological symptoms. Many people have trouble expressing internal emotions, consider mental illness to be a stigma, or simply assume depressive symptoms relate to their personal situations and therefore do not seek treatment. Physical symptoms are more prevalent among women, the elderly, the poor, children, culturally diverse populations, the medically ill, and the imprisoned. Because of a dual mechanism of action, medications such as duloxetine and venlafaxine may be useful in treating the physical symptoms as well as depressive symptoms in these special populations. PMID- 12755648 TI - The economic burden of depression with painful symptoms. AB - The economic burden of depression is substantial. The condition is highly prevalent, with both psychiatric and physical symptoms that often inflict pain. The chronic and often debilitating nature of depression results in costly medical therapies, as well as impaired workplace productivity. As a result, the overall economic burden of depression is comparable to that of serious physical illnesses, such as cancer and heart disease. This article presents an overview of the economic burden of depression and provides background on the relationship between depression and pain in this context. Research findings are also presented on the economic burden associated with a particular manifestation of pain among depressed patients, fibromyalgia. When painful physical symptoms accompany the already debilitating psychiatric and behavioral symptoms of depression, the economic burden that ensues for patients and their employers increases considerably. On purely economic grounds, more aggressive outreach may be warranted for patients with depression and comorbid pain to initiate treatment before symptoms are allowed to persist. However, more research is needed to assess the comprehensive economic impact that depression with painful physical symptoms can have on society. PMID- 12755649 TI - Depression with physical symptoms: treating to remission. AB - Depression is a recurrent, often chronic disease consisting of psychological and physical symptoms that are frequently undiagnosed or inadequately treated. While psychological symptoms have been shown to respond to current antidepressants, physical symptoms may not be as responsive. Treating both psychological and physical symptoms of depression may lead to a higher percentage of patients reaching remission. PMID- 12755650 TI - The ups and downs of novel antiemetic drugs, part 1: substance P, 5-HT, and the neuropharmacology of vomiting. AB - ISSUE: Novel antiemetic agents such as the newly approved aprepitant (Emend) target receptors for substance P, known as neurokinin-1 (NK(1)) receptors. When NK(1) receptors are blocked in the vomiting center of the brainstem, chemotherapy induced emesis is reduced. It is possible that blocking NK(1) receptors elsewhere in the CNS will lead to therapeutic actions in depression and other stress related disorders. PMID- 12755651 TI - A review of the safety and efficacy of droperidol for the rapid sedation of severely agitated and violent patients. AB - BACKGROUND: Droperidol had become a standard treatment for sedating severely agitated or violent patients in both psychiatric and medical emergency departments. However, several recent articles have suggested that droperidol may have a quinidine-like effect similar to that of thioridazine in inducing dysrhythmia. METHOD: In view of the recent U.S. Food and Drug Administration (FDA) position regarding the use of thioridazine, the authors reviewed the literature regarding droperidol and dysrhythmia in a MEDLINE search for the years 1960-2002 using the search terms droperidol, dysrhythmia, QTc interval, and sudden death as well as their own experience in using droperidol in a busy psychiatric emergency department. This review was done before the FDA's very recent and peremptory warning about droperidol. RESULTS: The authors report that, in treating approximately 12,000 patients over the past decade, they have never experienced a clinically significant adverse dysrhythmic event using droperidol to sedate severely agitated or violent patients. CONCLUSION: The authors conclude that, in clinical practice, droperidol is an extremely effective and safe method for treating severely agitated or violent patients. While in theory droperidol may prolong the QT interval to an extent similar to thioridazine, in clinical use there is no pattern of sudden deaths analogous to those that provoked the FDA warning about thioridazine. PMID- 12755652 TI - Factors associated with suicide attempts in 648 patients with bipolar disorder in the Stanley Foundation Bipolar Network. AB - BACKGROUND: Clinical factors related to suicide and suicide attempts have been studied much more extensively in unipolar depression compared with bipolar disorder. We investigated demographic and course-of-illness variables to better understand the incidence and potential clinical correlates of serious suicide attempts in 648 outpatients with bipolar disorder. METHOD: Patients with bipolar I or II disorder (DSM-IV criteria) diagnosed with structured interviews were evaluated using self-rated and clinician-rated questionnaires to assess incidence and correlates of serious suicide attempts prior to study entry. Clinician prospective ratings of illness severity were compared for patients with and without a history of suicide attempt. RESULTS: The 34% of patients with a history of suicide attempts, compared with those without such a history, had a greater positive family history of drug abuse and suicide (or attempts); a greater personal history of early traumatic stressors and more stressors both at illness onset and for the most recent episode; more hospitalizations for depression; a course of increasing severity of mania; more Axis I, II, and III comorbidities; and more time ill on prospective follow-up. In a hierarchical logistic regression, a history of sexual abuse, lack of confidant prior to illness onset, more prior hospitalizations for depression, suicidal thoughts when depressed, and cluster B personality disorder remained significantly associated with a serious suicide attempt. CONCLUSION: Our retrospective findings, supplemented by prospective follow-up, indicate that a history of suicide attempts is associated with a more difficult course of bipolar disorder and the occurrence of more psychosocial stressors at many different time domains. Greater attention to recognizing those at highest risk for suicide attempts and therapeutic efforts aimed at some of the correlates identified here could have an impact on bipolar illness-related morbidity and mortality. PMID- 12755653 TI - Residual symptoms in depression: can treatment be symptom-specific? AB - BACKGROUND: Most patients with depression continue to have symptoms after treatment. It is well documented that these "residual" symptoms are common and are associated with increases in suboptimal long-term outcomes such as relapse and disability. While it is clear that residual symptoms, as a group, contribute to poor outcomes, individual residual symptoms have received relatively little attention. To some extent, this lack of attention reflects an uncertainty in the field about the relationship of the syndrome of depression to the symptoms by which the syndrome is defined. METHOD: Recognizing that for clinicians and patients symptom relief is the goal of treatment, this article reviews the evidence that a symptomatic approach to individual residual symptoms is both feasible and useful. Evidence was gathered through a MEDLINE review of articles published in English from 1966 to 2002. Multiple keywords relating to symptoms, depression, and treatment were used. RESULTS: Many of the agents that psychiatrists use for augmentation of depression treatment, such as psychostimulants and alerting agents, atypical antipsychotics and mood stabilizers, and buspirone and benzodiazepines, have specific symptomatic effects, which raises the question of whether we are augmenting the core antidepressant effect or providing symptomatic relief. Fatigue, anxiety, sexual dysfunction, and sleep disturbances are all symptoms that are commonly leftover after treatment of depression. Some data indicate that treatment of these residual symptoms is efficacious and may affect the long-term outcome of depression. DISCUSSION: This discussion of the treatment of residual depressive symptoms raises a variety of research questions that should be addressed. Also implicit in this discussion are theoretical questions on the relationship between symptoms and syndrome. PMID- 12755654 TI - The effectiveness of quetiapine versus conventional antipsychotics in improving cognitive and functional outcomes in standard treatment settings. AB - OBJECTIVE: To examine the effectiveness of quetiapine versus conventional antipsychotics in improving cognitive and functional outcomes. METHOD: Forty stable outpatients with DSM-IV schizophrenia treated in public outpatient clinics were randomly assigned to continue taking conventional antipsychotic medications or switch to quetiapine for 6 months, beginning September 1998 and ending July 2000. Neurocognitive and functional measures were obtained at study entry, 3 months, and 6 months by raters blinded to treatment. Group differences were examined using repeated-measures analyses of covariance for mixed models. RESULTS: The mean (SD) dose of conventional antipsychotics in chlorpromazine equivalents was 348.00 (348.28) mg/day; the mean (SD) dose of quetiapine was 319.25 (142.55) mg/day. A cognitive function summary score improved in the quetiapine group relative to the group treated with conventional antipsychotics over the 6-month period (F = 5.80, df = 1,28.9; p <.023). Patients taking quetiapine did better with respect to both verbal fluency (initiation) and verbal memory. There were also statistically significant group differences with respect to quality of life favoring the quetiapine group (F = 4.87, df = 1,29; p <.04). Differences were not found with respect to adaptive functioning. CONCLUSION: Quetiapine improved cognition relative to conventional agents. After 6 months, groups differed by more than 1 standard deviation when baseline cognitive functioning was taken into account. No group differences were found with respect to improvements in community functioning. Improvements in adaptive functioning may lag behind improvements in cognition. Psychosocial programming may be necessary to translate gains in cognition into improvements in adaptive functioning. PMID- 12755655 TI - Safety of available agents used to treat bipolar disorder: focus on weight gain. AB - BACKGROUND: Pharmacotherapeutic management of bipolar disorder has advanced considerably since the introduction of lithium therapy nearly 50 years ago. The sizable percentage of patients who do not respond adequately to lithium and/or are intolerant to its side effects has served as an impetus for identifying alternative pharmacotherapeutic agents. Recent advances in the understanding of the neurotransmitter systems and their receptors as it applies to treatment of bipolar disorder has, in part, led to progress in delineating applications of anticonvulsant/antiepileptic drugs (AEDs) in this area. Although the efficacy of many drugs has been evaluated in patients with this disorder, medication tolerability and adherence issues related to unfavorable side effect profiles are substantial impediments to the development of novel pharmacotherapies. The potential for excessive weight gain as a side effect of certain psychopharmacologic agents remains a concern to both clinicians and patients. METHOD: English-language literature from 1985-2001 in MEDLINE was searched for the terms bipolar disorder, anticonvulsant, antiepileptic, lithium, antipsychotic, weight, and compliance. This article reviewed current therapeutic options for bipolar disorder, including newer AEDs and atypical antipsychotic drugs, with emphasis on the issue of weight gain and possible approaches to minimizing this risk. RESULTS: Certain newer AEDs are characterized by more favorable safety and tolerability profiles that include weight loss as a desirable side effect. Because bipolar disorder is associated with unacceptably high rates of relapse, recurrence, and morbidity, the concept of pharmacotherapeutic efficacy logically not only includes symptom relief but also necessarily encompasses issues related to regimen tolerability and adherence. CONCLUSION: There is a need for guidelines to help physicians carefully formulate and individualize management plans to reach safe, effective, and cost-efficient patient outcomes. Monitoring the weight of patients with bipolar disorder and educating them regarding this issue should be standard components of any treatment plan. PMID- 12755656 TI - Prevalence of hepatitis A, hepatitis B, and HIV among hepatitis C-seropositive state hospital patients: results from Oregon State Hospital. AB - BACKGROUND: Multiple studies have shown that individuals with severe mental illness are at increased risk for acquiring infection from human immunodeficiency virus (HIV), hepatitis B virus (HBV), and hepatitis C virus (HCV). Moreover, patients with chronic HCV infection are at risk for fulminant hepatitis from acquired infection with hepatitis A virus (HAV) or HBV, but there are limited data on the prevalence of HIV, HAV, and HBV in chronically hospitalized U.S. psychiatric patients without mental retardation who are HCV-seropositive. To address this issue, a comprehensive screening program was commenced at Oregon State Hospital (Salem, Ore.) beginning in 1999. METHOD: The computerized records of all non-geriatric adult inpatients at Oregon State Hospital on April 23, 2001, were reviewed to assess physician compliance with screening and the prevalence of infection with HIV, HAV, HBV, and HCV. RESULTS: Among the 535 patient records reviewed, 94.8% of patients were screened for HCV, of whom 20.3% were seropositive. Among HCV-seropositive patients, only 1.9% were not screened for HAV and HBV, but 23.3% were not tested for HIV. In the HCV-seropositive group, 35.9% were HAV-positive, 49.5% HBV-positive, and 2.6% HIV-positive. CONCLUSION: Chronic psychiatric inpatients have high HCV prevalence rates. Hepatitis C seropositive individuals may be at risk for complications unless vaccinated for HAV and HBV. PMID- 12755657 TI - Venlafaxine in treatment-resistant obsessive-compulsive disorder. AB - BACKGROUND: While selective serotonin reuptake inhibitors (SSRIs) are the first line treatment of obsessive-compulsive disorder (OCD), approximately 40% of patients fail to respond to SSRIs. Venlafaxine is a serotonin-norepinephrine reuptake inhibitor (SNRI) that might be effective in the treatment of OCD, even among those who have failed previous SSRI trials. METHOD: Thirty-nine patients who met DSM-IV criteria for OCD, including 29 who were resistant to prior SRI treatment trials, were treated with venlafaxine in an open, naturalistic fashion. Improvement was assessed using the Clinical Global Impressions-Improvement scale. RESULTS: Of 39 patients treated with venlafaxine, 27 (69.2%) were rated as sustained treatment responders. Of the 29 patients who did not respond to 1 or more previous SRI trials, 22 (75.9%) were rated as having sustained response to treatment. Mean dose of venlafaxine was 232.2 mg/day (range, 37.5-375 mg/day), and it was generally well tolerated. CONCLUSION: Venlafaxine may be beneficial to individuals with OCD, including those who have not responded to prior SSRI trials. However, these findings must be interpreted with caution, as the study is limited by its open, retrospective nature and its inclusion of patients with comorbid diagnoses and patients on concomitant medications. Prospective, controlled trials with a more homogeneous patient population are needed to replicate these preliminary findings. PMID- 12755658 TI - Pretreatment with ibuprofen to prevent electroconvulsive therapy-induced headache. AB - BACKGROUND: Although electroconvulsive therapy (ECT) has been widely recognized as an effective treatment for severe depression and various other psychiatric illnesses, adverse effects have been frequently reported, especially a high incidence of headache. Analgesics, such as acetaminophen, narcotics, or nonsteroidal anti-inflammatory drugs (NSAIDs), are commonly used to treat ECT induced headache. The objective of this study was to determine whether pretreatment with ibuprofen would prevent the onset or decrease the severity of headache that occurs after ECT. METHOD: All inpatients on the psychiatric units who required ECT treatment were asked to participate in the study. Thirty-four patients were randomly assigned to receive either ibuprofen, 600 mg, or placebo orally 90 minutes prior to the initial ECT session, with the alternate treatment given for the second ECT treatment. Patients were asked to complete a questionnaire prior to and after the first 2 ECT treatments regarding the pattern, severity, and onset of headache. Severity of the headache was measured on a visual analogue scale (VAS). RESULTS: Ten patients experienced headache in neither treatment arm, while 7 patients experienced headache in both treatment arms. Eleven patients experienced headache with placebo but not with ibuprofen, while 2 patients experienced headache with ibuprofen but not with placebo. Ibuprofen was significantly more effective than placebo in preventing the onset of headache post-ECT (p =.022). The mean +/- SD VAS headache scores were 1.49 +/- 1.54 and 0.54 +/- 0.91 in the placebo and ibuprofen arms, respectively. Ibuprofen was significantly more effective than placebo in reducing the severity of ECT induced headache (p =.007). CONCLUSION: Ibuprofen premedication reduced the frequency and severity of headache post-ECT and should be considered for appropriate patients who suffer from ECT-induced headache. PMID- 12755659 TI - Chronic depression and comorbid personality disorders: response to sertraline versus imipramine. AB - BACKGROUND: Chronic subtypes of depression appear to be associated with high rates of Axis II personality disorder comorbidity. Few studies, though, have systematically examined the clinical correlates of Axis II personality disorder comorbidity or its effect on treatment response or time to response. METHOD: 635 patients diagnosed with DSM-III-R chronic major depression or "double depression" (dysthymia with concurrent major depression) were randomized to 12 weeks of double-blind treatment with either sertraline or imipramine between February 1993 and December 1994. Axis II diagnoses were made using the personality disorders version of the DSM-III-R Structured Clinical Interview. The effect of study treatment was measured utilizing the Hamilton Rating Scale for Depression and the Clinical Global Impressions scale. RESULTS: Forty-six percent of patients met criteria for at least 1 comorbid Axis II personality disorder, with cluster C diagnoses being the most frequent at 39%; 21% met criteria for at least 2 Axis II personality disorders. A cluster C diagnosis was associated with significantly higher rates of early-onset depression (before age 21; 47% vs. 32% for no cluster C; p =.005) and comorbid anxiety disorder (34% vs. 18% for no cluster C; p <.001). Overall, the presence of Axis II personality disorder comorbidity had minimal-to-no effect on the ability to achieve either an antidepressant response or remission and had inconsistent effects on time to response. The presence of Axis II personality disorder comorbidity did not appear to reduce functional and quality-of-life improvements among patients responding to acute treatment with sertraline or imipramine. CONCLUSION: In this treatment sample, rates of Axis II personality disorder comorbidity were substantial in patients suffering from chronic forms of depression. Axis II personality disorder comorbidity did not appear to diminish symptomatic response to acute treatment or associated improvement in functioning and quality of life. PMID- 12755660 TI - Citalopram treatment of fluoxetine-intolerant depressed patients. AB - BACKGROUND: We assessed the tolerability of and response to citalopram in depressed patients who had discontinued fluoxetine treatment due to adverse events. METHOD: Fifty-five outpatients with DSM-IV major depressive disorder and a confirmed history of intolerance to fluoxetine (mean final dose = 24.6 mg/day) were switched to citalopram (20 mg/day) after a 2- to 4-week single-blind placebo washout period. During a 6-week, open-label treatment protocol, citalopram could be titrated up to 40 mg/day. Safety and tolerability, including reemergence of symptoms that previously had been associated with fluoxetine, were assessed by recording all spontaneously reported or observed adverse events. Efficacy was evaluated using the Hamilton Rating Scale for Depression (HAM-D), the Clinical Global Impressions (CGI) scale, and several other measures. Response was defined as a CGI-Improvement score at endpoint of 1 or 2 (i.e., very much or much improved). RESULTS: Ninety-five percent of patients (N = 52) completed the citalopram trial. The only adverse events reported by more than 5 patients (>or= 10%) were pharyngitis (15%) and constipation (11%), and none of the 3 early terminations were attributed to adverse events. The rate of recurrence of the fluoxetine-associated adverse events was low, with headache (3 [27%] of 11 cases), nausea (2 [22%] of 9 cases), and decreased libido (5 [18%] of 28 cases) being the most common. Significant improvement from baseline HAM-D (p <.001) was observed by the first week of citalopram therapy and continued until study end. The intent-to-treat CGI response rate was 65% (36 of 55 patients) at study endpoint; 69% (36 of 52 patients) of the completers responded. CONCLUSION: These data suggest that fluoxetine-intolerant patients can be treated effectively with citalopram. PMID- 12755661 TI - Conventional antipsychotic prescription in unipolar depression, I: an audit and recommendations for practice. AB - BACKGROUND: Despite narrow indications for conventional antipsychotics in depression, recent reports confirm a suspicion that they are widely prescribed in nonpsychotic depressive conditions. METHOD: Data from the case notes of over 510 patients with unipolar depression (unvalidated clinical diagnoses) were collected between June 1997 and January 1998 from community and acute units in 1 National Health Service (NHS) Trust. The aim of this audit was to assess the extent and pattern of antipsychotic prescription in this sample. RESULTS: More than a quarter (N = 138) of the sample (N = 494) were currently prescribed an antipsychotic; 40% of these received an antipsychotic without any recognized indication. The mean time on antipsychotic therapy was 3 years. Patients on antipsychotic therapy were, on average, taking twice as many total medications as those not on antipsychotic therapy. Patients with psychotic depression were taking an average of nearly twice the antipsychotic dose of nonpsychotic patients. CONCLUSION: Current clinical guidelines commend careful antidepressant choice in preference to polypharmacy. A number of drug choices for specific depressive presentations are summarized from recent sources. PMID- 12755662 TI - Metabolic syndrome in patients with schizophrenia. AB - BACKGROUND: Our aim was to study the frequency of metabolic syndrome and associated factors in patients with schizophrenia. METHOD: The study group consisted of 35 outpatients with long-term schizophrenia defined by DSM-IV criteria. Patients were assessed for the presence of metabolic syndrome, which was defined by the criteria of the National Cholesterol Education Program. All patients were on antipsychotic medication. Data were collected from Jan. 1, 2001, to Aug. 30, 2001. RESULTS: Metabolic syndrome was found in 37% (N = 13) of the patients, and it was associated inversely with the total daily dose of, but not with any specific type of, antipsychotic drug. CONCLUSION: The results suggest that metabolic syndrome is common among patients with schizophrenia, and it may be far more common than in general populations. PMID- 12755663 TI - Effectiveness of switching to ziprasidone for stable but symptomatic outpatients with schizophrenia. AB - BACKGROUND: Many outpatients with schizophrenia experience persistent symptoms or side effects on their current antipsychotic regimen. Few studies have prospectively examined the effects of the prior medication or switching method on the safety and efficacy of a newly available antipsychotic. Efficacy and tolerability of ziprasidone were evaluated in patients with DSM-IV schizophrenia or schizoaffective disorder who were switched from conventional or atypical antipsychotics in three 6-week, multicenter, randomized, open-label, parallel group trials. METHOD: Stable outpatients with persistent symptoms or troublesome side effects on (1) conventional antipsychotic (N = 108), (2) olanzapine (N = 104), or (3) risperidone (N = 58) therapy were switched to an open-label, 6-week, flexible-dose trial of ziprasidone (40-160 mg/day). Patients were randomly assigned at baseline to 1 of 3 switching schedules during the first week of ziprasidone therapy. Baseline and outcome assessments included Positive and Negative Syndrome Scale (PANSS) and Clinical Global Impressions of Severity (CGI S) ratings. RESULTS: All 3 switching strategies were well tolerated for all 3 patient groups. After 6 weeks on ziprasidone therapy, significant (p <.05) improvements were observed on all major symptom measures and almost all subscales for all switched subgroups. CONCLUSION: Switching stable but symptomatic outpatients from their previous antipsychotic to ziprasidone was generally well tolerated and was associated with symptom improvements 6 weeks later. Improvements occurred in patients recently on other first-line atypical antipsychotic, as well as in those on conventional antipsychotic, treatment. While limitations of switching study designs do not permit interpretation of comparative efficacy, these studies suggest that outpatients who partially respond to conventional antipsychotics, risperidone, or olanzapine may experience improved control of psychotic symptoms following a switch to ziprasidone. PMID- 12755664 TI - Risperidone compared with olanzapine in a naturalistic clinical study: a cost analysis. AB - BACKGROUND: Risperidone and olanzapine are thought to have broadly similar clinical effects. This study was designed as a cost analysis study comparing costs and basic clinical outcomes of treatment with risperidone or olanzapine in a naturalistic setting. METHOD: The U.K. Risperidone Olanzapine Drug Outcomes Studies in Schizophrenia (RODOS-UK) program consisted of a retrospective review of medical notes and prescription charts for 501 patients with schizophrenia or schizoaffective disorder who had been admitted to the hospital for the treatment of psychosis. The main outcome measure was cost of inpatient drug treatment. Clinical outcomes (clinician-assessed and -documented effectiveness, time to discharge) were also evaluated. Data were collected and verified between June and September 2000. RESULTS: Clinical outcomes were similar for risperidone and olanzapine. Clinician-assessed effectiveness was similar for both treatments (78% risperidone, 74% olanzapine; p =.39), but mean time to documented onset of effectiveness was significantly shorter for those treated with risperidone versus olanzapine (17.6 vs. 22.4 days; p =.01). Risperidone-treated patients stayed a mean of 9 fewer days in the hospital compared with olanzapine-treated patients (49 vs. 58 days; p =.007). The possibility that these observed differences were a result of different baseline characteristics could not be entirely discounted. Mean +/- SD doses of risperidone and olanzapine were 5.5 +/- 2.4 mg/day and 14.1 +/- 4.7 mg/day, respectively. The mean daily cost of all inpatient drugs was significantly higher for olanzapine than for risperidone (pound 5.63 vs. pound 3.92; p <.0001). Mean total costs of all inpatient drugs were significantly higher for olanzapine than for risperidone (pound 164 vs. pound 96; p <.0001), which partly reflected the longer mean treatment duration for olanzapine compared with risperidone (44 vs. 37 days). Concomitant antipsychotic use was similar for both groups (66% risperidone, 67% olanzapine). The number of patients documented as experiencing adverse events was not different between groups (22% risperidone, 19% olanzapine; p =.32). CONCLUSION: Risperidone and olanzapine produced broadly comparable clinical outcome in this cohort of hospitalized patients, but the use of risperidone was associated with significantly lower drug treatment costs. PMID- 12755665 TI - Serum leptin and triglyceride levels in patients on treatment with atypical antipsychotics. AB - BACKGROUND: Weight gain is a common adverse effect associated with the use of most antipsychotic drugs. Leptin has been reported to be associated with antipsychotic-induced weight gain. Previous studies have demonstrated a relationship between the atypical antipsychotics clozapine and olanzapine and serum leptin levels. We planned to comparatively investigate the effects of the atypical antipsychotics quetiapine, olanzapine, risperidone, and clozapine on leptin and triglyceride levels and weight gain. METHOD: The study population comprised 56 patients with DSM-IV schizophrenia, who were divided into 4 treatment groups: quetiapine (N = 14), olanzapine (N = 14), risperidone (N = 14), or clozapine (N = 14) monotherapy, and a control group of 11 patients receiving no psychopharmacologic treatment. The patients were evaluated at baseline and at the sixth week according to the Positive and Negative Syndrome Scale (PANSS), body mass index (BMI), weight, and fasting serum leptin and triglyceride levels. Data were gathered in 2001 and 2002. RESULTS: Olanzapine and clozapine caused a marked increase in weight and serum triglyceride and leptin levels, though increases in these variables were modest in the patients receiving quetiapine and minimal in those receiving risperidone. There were positive correlations between serum leptin levels and BMI and triglyceride levels. Clinical efficacy, as indicated by decrease in total PANSS scores, was associated with leptin levels in all atypical antipsychotic groups. CONCLUSION: Our results suggest that leptin may be associated with olanzapine- and clozapine-induced weight gain and that quetiapine appears to have modest influence and risperidone appears to have minimal influence on leptin and triglyceride levels and weight gain compared with olanzapine and clozapine. PMID- 12755666 TI - Undiagnosed hyperglycemia in clozapine-treated patients with schizophrenia. AB - BACKGROUND: Clozapine has been demonstrated to be superior to typical neuroleptics in reducing refractory symptoms in patients with schizophrenia, but it has also been associated with hyperglycemia and diabetes mellitus. This study was designed to investigate the proportion of undiagnosed impaired fasting glucose and diabetes mellitus in patients prescribed clozapine at 8 Department of Veterans Affairs (VA) medical centers. METHOD: All patients diagnosed by the VA in New England with ICD-9 schizophrenia from Oct. 1, 1999, to Sept. 30, 2000, who received a prescription for clozapine were identified, and an attempt was made to obtain a fasting plasma glucose (FPG) test. All patients were also characterized as to whether they were diagnosed as diabetic prior to the screening FPG. Patients not previously diagnosed as diabetic were divided into 2 groups: normal FPG (< 110 mg/dL) and elevated FPG (>or= 110 mg/dL). Clinical and sociodemographic characteristics of the 2 groups were compared using chi-square and t tests. RESULTS: Overall, 121 patients were not previously diagnosed as diabetic and received an FPG. Ninety-three (77%) had a normal FPG, and 28 (23%) had an elevated plasma glucose-including 17% with impaired fasting glucose and 6% with diabetes. Patients with hyperglycemia were significantly older (p =.007) and more commonly codiagnosed with bipolar disorder (p =.04). CONCLUSION: Hyperglycemia was common in patients receiving clozapine who had not been previously diagnosed as diabetic. These patients should be considered a group at high risk to develop diabetes mellitus and deserve both close monitoring and early intervention at the first sign of the onset of either diabetes or impaired glucose tolerance. PMID- 12755667 TI - Clozapine and venous thromboembolism: further evidence. PMID- 12755668 TI - Linezolid, a monoamine oxidase inhibiting antibiotic, and antidepressants. PMID- 12755669 TI - Olanzapine-sertraline combination in schizophrenia with obsessive-compulsive disorder. PMID- 12755670 TI - Two cases of alcohol craving curbed by topiramate. PMID- 12755671 TI - Quetiapine and QTc issues: a case report. PMID- 12755672 TI - The efficacy and safety of bilateral rTMS in medication-resistant depression. PMID- 12755673 TI - Racial differences in use of antipsychotics among patients with bipolar disorder. PMID- 12755675 TI - Developing a program of research in school nursing. PMID- 12755677 TI - Identifying and intervening with girls at risk for violence. AB - Youth violence has become a prominent national concern, largely focused on boys who have perpetrated highly publicized massacres. Less well-publicized is the rapid increase in arrests of girls for violent crimes and weapons violations. In just 2 decades, violent crime arrests for female juveniles increased by 108%. From research findings, a composite portrait of the violent girl can be drawn. This profile can be used to identify girls at risk for criminal behaviors. In this article, a three-pronged approach to girls' violent behavior is presented: (a) violence prevention and emotional literacy programs that can be implemented by school nurses, (b) parent education programs that can be conducted at Parent Teacher Association meetings, churches, and community centers, and (c) counseling interventions that can be delivered to troubled girls by psychiatric nurses. Nurses can play vital roles in consultation to teachers and parents and in direct service provision to girls who are on a tragic trajectory of fighting, expulsion from school, and juvenile justice infractions. PMID- 12755678 TI - The importance of hearing conservation instruction. AB - According to Denehy (1999), "school nurses can play a powerful role in promoting health in their schools and community." She encouraged school nurses to "Take and make opportunities to promote health in the classroom" (p. 4). Classroom presentation of hearing conservation information is one way for school nurses to promote health and to reduce the prevalence of noise-induced hearing loss (NIHL), an irreversible yet preventable condition. Because of excessive sound exposure, the prevalence of NIHL among children is increasing. Numerous experts have recommended the implementation of hearing conservation education programs in schools. Despite these recommendations made over the last 3 decades, basic hearing conservation information that could prevent countless cases of NIHL remains conspicuously absent from most school curricula. School nurses should seize this "golden opportunity" to promote health in the classroom and help to reduce the prevalence of NIHL. PMID- 12755679 TI - Recommended minimal emergency equipment and resources for schools: national consensus group report. AB - Providing an environment that is responsive to emergency health needs of students is essential to creating a safe setting for children in schools. The question of what minimal essential emergency equipment and resources should be available in schools brings with it many and varied opinions, issues, and concerns. Through funding from the Emergency Medical Services for Children (EMSC), the National Association of School Nurses (NASN) was charged with the task of convening a consensus group to formulate a recommended list of minimal essential emergency equipment and resources that should be present in all schools. This article provides an overview of the issues surrounding minimal emergency equipment needs for schools, presents recommended minimal emergency equipment and resources, and recommendations for further actions. PMID- 12755680 TI - Quality assurance in school health. AB - School nurses need to become more influential administrators, managers, and entrepreneurs. They must learn to lead and collaborate effectively in designing, implementing, and evaluating coordinated school health programs. Quality assurance is an essential ingredient in this process that requires accurate, timely, and confidential incident reporting and data analysis structures. These structures, in turn, can serve as the foundation of evidence-based practice and overall system improvement. School nurses can and should assume a key role in the process and thereby continue to meet the challenges of the more sophisticated school health services that today's student population requires. In this way, school nurses will continue to protect and advance the health and safety of the students who are entrusted to their care. PMID- 12755681 TI - The impact of school nursing on school performance: a research synthesis. AB - School nurses work in an educational setting. Due to budget cuts, different goals, and confusion between educators and nurses regarding the school nurse role, school nurses are being asked to demonstrate their effectiveness and justify their presence in elementary and secondary schools. Although school nursing was first initiated 100 years ago, a review of the literature published since 1965 indicates that 15 studies have been conducted that examine the impact of school nurses on academic performance. However, today many articles have recommended more research linking school nursing to educational outcomes. This article synthesizes the results of 15 research articles. Findings from these studies indicate that nursing interventions targeted at specific populations, including parents, have had significant effects. A relationship between school nurses' interventions and absenteeism is also suggested. Limitations of these studies are discussed, along with suggestions for future research. PMID- 12755682 TI - Measures of overweight status in school-age children. AB - Identifying and intervening with overweight children may decrease their likelihood of developing heart disease later in life. This secondary analysis of 58 children in the 3rd grade examined the prevalence of overweight children, methods for measuring overweight status, and the relationship among these measures and other risk factors for heart disease. Approximately one third of the 58 children were categorized as overweight. Several measures, such as weight, body fat percentage, body mass index (BMI), and skin-fold, are available to school nurses for measuring overweight status. The highest correlations were between BMI and weight and between BMI and body fat. Anthropometric measurements cannot predict cholesterol level, 24-hour diet recall, or family history. Blood pressure can be predicted by weight, body fat percentage, and BMI. BMI and body fat percentage highly correlate; however, body fat percentage is more liberal in identifying children at risk for overweight status. Therefore, body fat percentage is recommended for identification of overweight status in school-age children. PMID- 12755683 TI - Using active learning strategies to present bloodborne pathogen programs. AB - Every year, school nurses have the responsibility for developing and presenting a bloodborne pathogen presentation to the education and clerical staff of their buildings. Although the information is similar from year to year, the manner in which the information is presented can be altered. Teachers are using active learning strategies in a variety of learning environments, engaging students in the learning process by having them play an active role. With some planning, preparation, and imagination, active learning strategies can be incorporated into bloodborne pathogen presentations. The purpose of this article is to define active learning, describe how to develop a program using active learning strategies, and provide some examples of bloodborne pathogen presentations that have already been developed. Several sources are identified that can provide the school nurse with information regarding bloodborne pathogens. Information about how computers can be integrated into the bloodborne pathogen presentation is also presented. PMID- 12755684 TI - TIMP-2 (tissue inhibitor of metalloproteinase-2) regulates MMP-2 (matrix metalloproteinase-2) activity in the extracellular environment after pro-MMP-2 activation by MT1 (membrane type 1)-MMP. AB - The matrix metalloproteinase (MMP)-2 has a crucial role in extracellular matrix degradation associated with cancer metastasis and angiogenesis. The latent form, pro-MMP-2, is activated on the cell surface by the membrane-tethered membrane type 1 (MT1)-MMP, in a process regulated by the tissue inhibitor of metalloproteinase (TIMP)-2. A complex of active MT1-MMP and TIMP-2 binds pro-MMP 2 forming a ternary complex, which permits pro-MMP-2 activation by a TIMP-2-free neighbouring MT1-MMP. It remains unclear how MMP-2 activity in the pericellular space is regulated in the presence of TIMP-2. To address this question, the effect of TIMP-2 on MMP-2 activity in the extracellular space was investigated in live cells, and their isolated plasma membrane fractions, engineered to control the relative levels of MT1-MMP and TIMP-2 expression. We show that both free and inhibited MMP-2 is detected in the medium, and that the net MMP-2 activity correlates with the level of TIMP-2 expression. Studies to displace MT1-MMP-bound TIMP-2 in a purified system with active MMP-2 show minimal displacement of inhibitor, under the experimental conditions, due to the high affinity interaction between TIMP-2 and MT1-MMP. Thus inhibition of MMP-2 activity in the extracellular space is unlikely to result solely as a result of TIMP-2 dissociation from its complex with MT1-MMP. Consistently, immunoblot analyses of plasma membranes, and surface biotinylation experiments show that the level of surface association of TIMP-2 is independent of MT1-MMP expression. Thus low affinity binding of TIMP-2 to sites distinct to MT1-MMP may have a role in regulating MMP-2 activity in the extracellular space generated by the ternary complex. PMID- 12755685 TI - Protein S-thiolation targets glycolysis and protein synthesis in response to oxidative stress in the yeast Saccharomyces cerevisiae. AB - The irreversible oxidation of cysteine residues can be prevented by protein S thiolation, a process by which protein SH groups form mixed disulphides with low molecular-mass thiols such as glutathione. We report here the target proteins which are modified in yeast cells in response to H(2)O(2). In particular, a range of glycolytic and related enzymes (Tdh3, Eno2, Adh1, Tpi1, Ald6 and Fba1), as well as translation factors (Tef2, Tef5, Nip1 and Rps5) are identified. The oxidative stress conditions used to induce S-thiolation are shown to inhibit GAPDH (glyceraldehyde-3-phosphate dehydrogenase), enolase and alcohol dehydrogenase activities, whereas they have no effect on aldolase, triose phosphate isomerase or aldehyde dehydrogenase activities. The inhibition of GAPDH, enolase and alcohol dehydrogenase is readily reversible once the oxidant is removed. In addition, we show that peroxide stress has little or no effect on glucose-6-phosphate dehydrogenase or 6-phosphogluconate dehydrogenase, the enzymes that catalyse NADPH production via the pentose phosphate pathway. Thus the inhibition of glycolytic flux is proposed to result in glucose equivalents entering the pentose phosphate pathway for the generation of NADPH. Radiolabelling is used to confirm that peroxide stress results in a rapid and reversible inhibition of protein synthesis. Furthermore, we show that glycolytic enzyme activities and protein synthesis are irreversibly inhibited in a mutant that lacks glutathione, and hence cannot modify proteins by S-thiolation. In summary, protein S-thiolation appears to serve an adaptive function during exposure to an oxidative stress by reprogramming metabolism and protecting protein synthesis against irreversible oxidation. PMID- 12755686 TI - The transcription factor early growth response factor-1 (EGR-1) promotes apoptosis of neuroblastoma cells. AB - Early growth response factor-1 (EGR-1) is an immediate early gene, which is rapidly activated in quiescent cells by mitogens or in postmitotic neurons after depolarization. EGR-1 has been involved in diverse biological functions such as cell growth, differentiation and apoptosis. Here we report that enforced expression of the EGR-1 gene induces apoptosis, as determined by flow cytometry and terminal deoxynucleotidyl transferase-mediated dUTP-fluorescein nick-end labelling (TUNEL) analysis, in murine Neuro2A cells. In accordance with this role of EGR-1 in cell death, antisense oligonucleotides increase cell viability in cells cultured in the absence of serum. This apoptotic activity of the EGR-1 appears to be mediated by p73, a member of the p53 family of proteins, since an increase in the amount of p73 is observed in clones stably expressing the EGR-1 protein. We also observed an increase in the transcriptional activity of the mdm2 promoter in cells overexpressing EGR-1, which is paralleled by a marked decrease in the levels of p53 protein, therefore excluding a role of this protein in mediating EGR-1-induced apoptosis. Our results suggest that EGR-1 is an important factor involved in neuronal apoptosis. PMID- 12755687 TI - Characterization of the interaction between alpha2-macroglobulin and fibroblast growth factor-2: the role of hydrophobic interactions. AB - Basic fibroblast growth factor (FGF-2) is important in development, wound healing and angiogenesis. The human plasma proteinase inhibitor alpha2-macroglobulin (alpha2M) binds to and regulates the biological activity of various growth factors, including FGF-2. FGF-2 binds specifically and saturably to native alpha2M and conformationally modified alpha2M (alpha2M*); however, the KD for FGF 2 binding to alpha2M* is 10-fold lower. This study investigates the biochemical nature of the interaction between FGF-2 and alpha2M* and localizes a possible FGF 2 binding site in the alpha2M subunit. FGF-2 binding to alpha2M* was not affected by shifts in pH between 6.5 and 10; however, increasing temperature decreased the KD for this interaction. The binding affinity of FGF-2 for alpha2M* also increased with increasing ionic strength. These results are consistent with the hypothesis that hydrophobic interactions predominate in promoting FGF-2 association with alpha2M*. Consistent with this hypothesis, FGF-2 bound to a glutathione S-transferase fusion protein containing amino acids 591-774 of the alpha2M subunit (FP3) and to a hydrophobic 16-amino-acid peptide (amino acids 718 733) within FP3. Specific binding of FGF-2 to the 16-amino-acid peptide was inhibited by excess transforming growth factor-beta1. When the 16-amino-acid peptide was chemically modified to neutralize the only two charged amino acids, FGF-2-binding activity was unaffected, supporting the predominant role of hydrophobic interactions. FGF-2 presentation to signalling receptors is influenced by growth factor binding to heparan sulphate proteoglycans (HSPGs), which is electrostatic in nature. Our results demonstrate that the interactions of FGF-2 with alpha2M* and HSPGs are biochemically distinct, suggesting that different FGF-2 sequences are involved. PMID- 12755689 TI - Detection and characterization of a novel extracellular fungal enzyme that catalyzes the specific and hydrolytic cleavage of lignin guaiacylglycerol beta aryl ether linkages. AB - Cleavage of the arylglycerol beta-aryl ether linkage is the most important process in the biological degradation of lignin. The bacterial beta-etherase was described previously and shown to be tightly associated with the cellular membrane. In this study, we aimed to detect and isolate a new extracellular function that catalyses the beta-aryl ether linkage cleavage of high-molecular lignin in the soil fungi. We screened and isolated 2BW-1 cells by using a highly sensitive fluorescence assay system. The beta-aryl ether cleavage enzyme was produced by a newly isolated fungus, 2BW-1, and is secreted into the extracellular fraction. The beta-aryl ether cleavage enzyme converts the guaiacylglycerol beta-O-guaiacyl ether (GOG) to guaiacylglycerol and guaiacol. It requires the C alpha alcohol structure and p-hydroxyl group and specifically attacks the beta-aryl ether linkage of high-molecular mass lignins with addition of two water molecules at the C alpha and C beta positions. PMID- 12755688 TI - The Theodore Bucher lecture. Investigating signal transduction with genetically encoded fluorescent probes. AB - Ca2+ and cAMP are ubiquitous second messengers in eukaryotes and control numerous physiological responses ranging from fertilization to cell death induction. To distinguish between these different responses, their subtle regulation in time, space and amplitude is needed. Therefore, the characterization of the signalling process requires measurement of second messengers with tools of precise localization, high dynamic range and as little disturbance of cell physiology as possible. Recently, fluorescent proteins of marine jellyfish have given rise to a set of genetically encoded biosensors which fulfil these criteria and which have already led to important new insights into the subcellular handling of Ca2+ and cAMP. The use of these probes in combination with new microscopical methods such as two-photon microscopy now enables researchers to study second messenger signalling in intact tissues. In this review, the genetically encoded measurement probes and their origin are briefly introduced and some recent insights into the spatio-temporal complexity of both Ca2+ and cAMP signalling obtained with these tools are discussed. PMID- 12755690 TI - Thyroid Ca2+/NADPH-dependent H2O2 generation is partially inhibited by propylthiouracil and methimazole. AB - H2O2 generation is a limiting step in thyroid hormone biosynthesis. Biochemical studies have confirmed that H2O2 is generated by a thyroid Ca2+/NADPH-dependent oxidase. Decreased H2O2 availability may be another mechanism of inhibition of thyroperoxidase activity produced by thioureylene compounds, as propylthiouracil (PTU) and methimazole (MMI) are antioxidant agents. Therefore, we analyzed whether PTU or MMI could scavenge H2O2 or inhibit thyroid NADPH oxidase activity in vitro. Our results show that PTU and thiourea did not significantly scavenge H2O2. However, MMI significantly scavenged H2O2 at high concentrations. Only MMI was able to decrease the amount of H2O2 generated by the glucose-glucose oxidase system. On the other hand, both PTU and MMI were able to partially inhibit thyroid NADPH oxidase activity in vitro. As PTU did not scavenge H2O2 under the conditions used here, we presume that this drug may directly inhibit thyroid NADPH oxidase. Also, at the concentration necessary to inhibit NADPH oxidase activity, MMI did not scavenge H2O2, also suggesting a direct effect of MMI on thyroid NADPH oxidase. In conclusion, this study shows that MMI, but not PTU, is able to scavenge H2O2 in the micromolar range and that both PTU and MMI can impair thyroid H2O2 generation in addition to their potent thyroperoxidase inhibitory effects. PMID- 12755691 TI - Investigation of the kinetics and order of tyrosine phosphorylation in the T-cell receptor zeta chain by the protein tyrosine kinase Lck. AB - We report experiments to investigate the role of the physiologically relevant protein tyrosine kinase Lck in the ordered phosphorylation of the T-cell receptor zeta chain. Six synthetic peptides were designed based on the sequences of the immunoreceptor tyrosine-based activation motifs (ITAMs) of the zeta chain. Preliminary 1H-NMR studies of recombinant zeta chain suggested that it is essentially unstructured and therefore that peptide mimics would serve as useful models for investigating individual ITAM tyrosines. Phosphorylation kinetics were determined for each tyrosine by assaying the transfer of 32P by recombinant Lck on to each of the peptides. The rates of phosphorylation were found to depend on the location of the tyrosine, leading to the proposal that Lck phosphorylates the six zeta chain ITAM tyrosines in the order 1N (first) > 3N > 3C > 2N > 1C > 2C (last) as a result of differences in the amino-acid sequence surrounding each tyrosine. This proposal was then tested on cytosolic, recombinant T-cell receptor zeta chain. After in vitro phosphorylation by Lck, the partially phosphorylated zeta chain was digested with trypsin. Separation and identification of the zeta chain fragments using LC-MS showed, as predicted by the peptide phosphorylation studies, that tyrosine 1N is indeed the first to be phosphorylated by Lck. We conclude that differences in the amino-acid context of the six zeta chain ITAM tyrosines affect the efficiency of their phosphorylation by the kinase Lck, which probably contributes to the distinct patterns of phosphorylation observed in vivo. PMID- 12755692 TI - Selectivity of pyruvate kinase for Na+ and K+ in water/dimethylsulfoxide mixtures. AB - In aqueous media, muscle pyruvate kinase is highly selective for K+ over Na+. We now studied the selectivity of pyruvate kinase in water/dimethylsulfoxide mixtures by measuring the activation and inhibition constants of K+ and Na+, i.e. their binding to the monovalent and divalent cation binding sites of pyruvate kinase, respectively [Melchoir J.B. (1965) Biochemistry 4, 1518-1525]. In 40% dimethylsulfoxide the K0.5 app for K+ and Na+ were 190 and 64-fold lower than in water. Ki app for K+ and Na+ decreased 116 and 135-fold between 20 and 40% dimethylsulfoxide. The ratios of Ki app/K0.5 app for K+ and Na+ were 34-3.5 and 3.3-0.2, respectively. Therefore, dimethylsulfoxide favored the partition of K+ and Na+ into the monovalent and divalent cation binding sites of the enzyme. The kinetics of the enzyme at subsaturating concentrations of activators show that K+ and Mg2+ exhibit high selectivity for their respective cation binding sites, whereas when Na+ substitutes K+, Na+ and Mg2+ bind with high affinity to their incorrect sites. This is evident by the ratio of the affinities of Mg2+ and K+ for the monovalent cation binding site, which is close to 200. For Na+ and Mg2+ this ratio is approximately 20. Therefore, the data suggest that K+ induces conformational changes that prevent the binding of Mg2+ to the monovalent cation binding site. Circular dichroism spectra of the enzyme and the magnitude of the transfer and apparent binding energies of K+ and Na+ indicate that structural arrangements of the enzyme induced by dimethylsulfoxide determine the affinities of pyruvate kinase for K+ and Na+. PMID- 12755693 TI - Tumor necrosis factor-alpha converting enzyme is processed by proprotein convertases to its mature form which is degraded upon phorbol ester stimulation. AB - Tumor necrosis factor-alpha converting enzyme (TACE or ADAM17) is a member of the ADAM (a disintegrin and metalloproteinase) family of type I membrane proteins and mediates the ectodomain shedding of various membrane-anchored signaling and adhesion proteins. TACE is synthesized as an inactive zymogen, which is subsequently proteolytically processed to the catalytically active form. We have identified the proprotein-convertases PC7 and furin to be involved in maturation of TACE. This maturation is negatively influenced by the phorbol ester phorbol-12 myristate-13-acetate (PMA), which decreases the cellular amount of the mature form of TACE in PMA-treated HEK293 and SH-SY5Y cells. Furthermore, we found that stimulation of protein kinase C or protein kinase A signaling pathways did not influence long-term degradation of mature TACE. Interestingly, PMA treatment of furin-deficient LoVo cells did not affect the degradation of mature TACE. By examination of furin reconstituted LoVo cells we were able to exclude the possibility that PMA modulates furin activity. Moreover, the PMA dependent decrease of the mature enzyme form is specific for TACE, as the amount of mature ADAM10 was unaffected in PMA-treated HEK293 and SH-SY5Y cells. Our results indicate that the activation of TACE by the proprotein-convertases PC7 and furin is very similar to the maturation of ADAM10 although there is a significant difference in the cellular stability of the mature enzyme forms after phorbol ester treatment. PMID- 12755694 TI - Selective inhibition of ADAMTS-1, -4 and -5 by catechin gallate esters. AB - Three mammalian ADAMTS enzymes, ADAMTS-1, -4 and -5, are known to cleave aggrecan at certain glutamyl bonds and are considered to be largely responsible for cartilage aggrecan catabolism observed during the development of arthritis. We have previously reported that certain catechins, polyphenolic compounds found in highest concentration in green tea (Camellia sinensis), are capable of inhibiting cartilage aggrecan breakdown in an in vitro model of cartilage degradation. We have now cloned and expressed recombinant human ADAMTS-1, -4 and -5 and report here that the catechin gallate esters found in green tea potently inhibit the aggrecan-degrading activity of these enzymes, with submicromolar IC50 values. Moreover, the concentration needed for total inhibition of these members of the ADAMTS group is approximately two orders of magnitude lower than that which is needed to partially inhibit collagenase or ADAM-10 activity. Catechin gallate esters therefore provide selective inhibition of certain members of the ADAMTS group of enzymes and could constitute an important nutritional aid in the prevention of arthritis as well as being part of an effective therapy in the treatment of joint disease and other pathologies involving the action of these enzymes. PMID- 12755695 TI - Sulfation of hydroxychlorobiphenyls. Molecular cloning, expression, and functional characterization of zebrafish SULT1 sulfotransferases. AB - As a first step toward developing a zebrafish model for investigating the role of sulfation in counteracting environmental estrogenic chemicals, we have embarked on the identification and characterization of cytosolic sulfotransferases (STs) in zebrafish. By searching the zebrafish expressed sequence tag database, we have identified two cDNA clones encoding putative cytosolic STs. These two zebrafish ST cDNAs were isolated and subjected to nucleotide sequencing. Sequence data revealed that the two zebrafish STs are highly homologous, being approximately 82% identical in their amino acid sequences. Both of them display approximately 50% amino acid sequence identity to human SULT1A1, rat SULT1A1, and mouse SULT1C1 ST. These two zebrafish STs therefore appear to belong to the SULT1 cytosolic ST gene family. Recombinant zebrafish STs (designated SULT1 STs 1 and 2), expressed using the pGEX-2TK prokaryotic expression system and purified from transformed Escherichia coli cells, migrated as approximately 35 kDa proteins on SDS/PAGE. Purified zebrafish SULT1 STs 1 and 2 displayed differential sulfating activities toward a number of endogenous compounds and xenobiotics including hydroxychlorobiphenyls. Kinetic constants of the two enzymes toward two representative hydroxychlorobiphenyls, 3-chloro-4-biphenylol and 3,3',5,5' tetrachloro-4,4'-biphenyldiol, and 3,3',5-triiodo-l-thyronine were determined. A thermostability experiment revealed the two enzymes to be relatively stable over the range 20-43 degrees C. Among 10 different divalent metal cations tested, Co2+, Zn2+, Cd2+, and Pb2+ exhibited considerable inhibitory effects, while Hg2+ and Cu2+ rendered both enzymes virtually inactive. PMID- 12755696 TI - Differential post-translational modification of CD63 molecules during maturation of human dendritic cells. AB - The capacity of dendritic cells to initiate T cell responses is related to their ability to redistribute MHC class II molecules from the intracellular MHC class II compartments to the cell surface. This redistribution occurs during dendritic cell development as they are converted from an antigen capturing, immature dendritic cell into an MHC class II-peptide presenting mature dendritic cell. During this maturation, antigen uptake and processing are down-regulated and peptide-loaded class II complexes become expressed in a stable manner on the cell surface. Here we report that the tetraspanin CD63, that associates with intracellularly localized MHC class II molecules in immature dendritic cells, was modified post-translationally by poly N-acetyl lactosamine addition during maturation. This modification of CD63 was accompanied by a change in morphology of MHC class II compartments from typical multivesicular organelles to structures containing densely packed lipid moieties. Post-translational modification of CD63 may be involved in the functional and morphological changes of MHC class II compartments that occur during dendritic cell maturation. PMID- 12755697 TI - Comparative analysis of the ATP-binding sites of Hsp90 by nucleotide affinity cleavage: a distinct nucleotide specificity of the C-terminal ATP-binding site. AB - The 90-kDa heat shock protein (Hsp90) is a molecular chaperone that assists both in ATP-independent sequestration of damaged proteins, and in ATP-dependent folding of numerous targets, such as nuclear hormone receptors and protein kinases. Recent work from our lab and others has established the existence of a second, C-terminal nucleotide binding site besides the well characterized N terminal, geldanamycin-sensitive ATP-binding site. The cryptic C-terminal site becomes open only after the occupancy of the N-terminal site. Our present work demonstrates the applicability of the oxidative nucleotide affinity cleavage in the site-specific characterization of nucleotide binding proteins. We performed a systematic analysis of the nucleotide binding specificity of the Hsp90 nucleotide binding sites. N-terminal binding is specific to adenosine nucleotides with an intact adenine ring. Nicotinamide adenine dinucleotides and diadenosine polyphosphate alarmones are specific N-terminal nucleotides. The C-terminal binding site is much more unspecific-it interacts with both purine and pirimidine nucleotides. Efficient binding to the C-terminal site requires both charged residues and a larger hydrophobic moiety. GTP and UTP are specific C-terminal nucleotides. 2',3'-O-(2,4,6-trinitrophenyl)-nucleotides (TNP-ATP, TNP-GTP) and pyrophosphate access the C-terminal binding site without the need for an occupied N-terminal site. Our data provide additional evidence for the dynamic domain domain interactions of Hsp90, give hints for the design of novel types of specific Hsp90 inhibitors, and raise the possibility that besides ATP, other small molecules might also interact with the C-terminal nucleotide binding site in vivo. PMID- 12755698 TI - Characterization of a high-affinity binding site for the pea albumin 1b entomotoxin in the weevil Sitophilus. AB - The toxicity of the pea albumin 1b (PA1b), a 37 amino-acid peptide extracted from pea seeds, for cereal weevils (Sitophilus oryzae, Sitophilus granarius and Sitophilus zeamais) was recently discovered. The mechanism of action of this new entomotoxin is still unknown and potentially involves a target protein in the insect tissues. This work describes the characterization of a high-affinity binding site for PA1b in a microsomal fraction of Sitophilus spp. extracts. Purified PA1b was labeled to a high specific radioactivity (c. 900 Ci.mmol-1) using 125I, and the iodinated ligand was found to be biologically active. Binding of this ligand to the microsomal fraction of S. oryzae extract was found to be saturable and reversible, with an affinity (Kd) of 2.6 nm, and a high maximal binding capacity (Bmax) of 40 pmol.mg-1 of protein. A binding site displaying similar characteristics was detectable in the five susceptible weevils strains tested, as well as in the pea aphid or in the fruit fly. However, no binding activity was detectable in extracts from four S. oryzae strains previously shown to be resistant to the toxin through a recessive monogenic mechanism. Therefore, we suggest that this binding site might be involved in the mechanism of action of PA1b. PMID- 12755699 TI - A novel retinol-binding protein in the retina of the swallowtail butterfly, Papilio xuthus. AB - Retinoid-binding proteins are indispensable for visual cycles in both vertebrate and invertebrate retinas. These proteins stabilize and transport hydrophobic retinoids in the hydrophilic environment of plasma and cytoplasm, and allow regeneration of visual pigments. Here, we identified a novel retinol-binding protein in the eye of a butterfly, Papilio xuthus. The protein that we term Papilio retinol-binding protein (Papilio RBP) is a major component of retinal soluble proteins and exclusively binds 3-hydroxyretinol, and emits fluorescence peaking at 480 nm under ultraviolet (UV) illumination. The primary structure, deduced from the nucleotide sequence of the cDNA, shows no similarity to any other lipophilic ligand-binding proteins. The molecular mass and isoelectric point of the protein estimated from the amino-acid sequence are 26.4 kDa and 4.92, respectively. The absence of any signal sequence for secretion in the N terminus suggests that the protein exists in the cytoplasmic matrix. All-trans 3 hydroxyretinol is the major ligand of the Papilio RBP in dark-adapted eyes. Light illumination of the eyes increases the 11-cis isomer of the ligand and induces redistribution of the Papilio RBP from the proximal to the distal part of the photoreceptor layer. These results suggest that the Papilio RBP is involved in visual pigment turnover. PMID- 12755700 TI - Reversed-phase HPLC determination of chlorophyll a' and phylloquinone in Photosystem I of oxygenic photosynthetic organisms. Universal existence of one chlorophyll a' molecule in Photosystem I. AB - Chlorophyll (Chl) a', the C132-epimer of Chl a, is a constituent of the primary electron donor (P700) of Photosystem (PS) I of a thermophilic cyanobacterium Synechococcus (Thermosynechococcus) elongatus, as was recently demonstrated by X ray crystallography. To determine whether PS I of oxygenic photosynthetic organisms universally contains one molecule of Chl a', pigment compositions of thylakoid membranes and PS I complexes isolated from the cyanobacteria T. elongatus and Synechocystis sp. PCC 6803, the green alga Chlamydomonas reinhardtii, and the green plant spinach, were examined by simultaneous detection of phylloquinone (the secondary electron acceptor of PS I) and Chl a' by reversed phase HPLC. The results were compared with the Chl a/P700 ratio determined spectrophotometrically. The Chl a'/PS I ratios of thylakoid membranes and PS I were about 1 for all the organisms examined, and one Chl a' molecule was found in PS I even after most of the peripheral subunits were removed. Chl a' showed a characteristic extraction behaviour significantly different from the bulk Chl a in acetone/methanol extraction upon varying the mixing ratio. These findings confirm that a single Chl a' molecule in P700 is the universal feature of PS I of the Chl a-based oxygenic photosynthetic organisms. PMID- 12755701 TI - Emerin interacts in vitro with the splicing-associated factor, YT521-B. AB - Emerin is a nuclear membrane protein that interacts with lamin A/C at the nuclear envelope. Mutations in either emerin or lamin A/C cause Emery-Dreifuss muscular dystrophy (EDMD). The functions of emerin are poorly understood, but EDMD affects mainly skeletal and cardiac muscle. We used a high-stringency yeast two-hybrid method to screen a human heart cDNA library, with full-length emerin as bait. Four out of five candidate interactors identified were nuclear proteins: lamin A, splicing factor YT521-B, proteasome subunit PA28 gamma and transcription factor vav-1. Specific binding between emerin and the functional C-terminal domain of YT521-B was confirmed by pull-down assays and biomolecular interaction analysis (BIAcore). Inhibition by emerin of YT521-B-dependent splice site selection in vivo suggests that the interaction is physiologically significant. A 'bipartite' binding site for YT521-B in emerin was identified using alanine substitution or disease-associated mutations in emerin. The transcription factor GCL (germ cell less) has previously been shown to bind to the same site. The results are consistent with an emerging view that lamins and lamina-associated proteins, like emerin, have a regulatory role, as well as a structural role in the nucleus. YT521-B joins a growing list of candidates for a role in a gene expression model of the pathogenesis of EDMD. PMID- 12755702 TI - The Hansenula polymorpha MOX gene presents two alternative transcription start points differentially utilized and sensitive to respiratory activity. AB - The peroxisomal methanol metabolism of Hansenula polymorpha depends on a group of genes that are coordinately regulated. Methanol oxidase (Mox) plays a key role in this pathway and its synthesis has been shown to be regulated at the transcriptional level. MOX expression is strongly repressed on glucose and activated on glycerol or methanol. In this study we have identified two MOX transcripts that are differentially expressed along MOX derepression. The first one, named l-MOX (for longer MOX), starts at position -425, is only weakly and transiently transcribed and is not translated into the Mox protein. The other is the true MOX mRNA, which initiates around position -25. Using a strain bearing multiple copies of MOX(Q1N) and a reporter gene fused to the MOX promoter, regulation of the two transcripts was investigated. Initiation of the true MOX correlates with repression of l-MOX and conditions that are repressive for MOX transcription, such as the inhibition of mitochondrial activity, lead to higher levels of l-MOX expression. This effect was first observed in a mox mutant (Q1N M8) unable to grow on nonfermentable carbon sources. No function was detected for l-MOX, but its regulation follows a pattern similar to that of catalase, which is essential for methanol metabolism. This suggests that, l-MOX, although precisely regulated, seems to be a remnant of the evolution of the methanol metabolism network. PMID- 12755703 TI - Two W-containing formate dehydrogenases (CO2-reductases) involved in syntrophic propionate oxidation by Syntrophobacter fumaroxidans. AB - Two formate dehydrogenases (CO2-reductases) (FDH-1 and FDH-2) were isolated from the syntrophic propionate-oxidizing bacterium Syntrophobacter fumaroxidans. Both enzymes were produced in axenic fumarate-grown cells as well as in cells which were grown syntrophically on propionate with Methanospirillum hungatei as the H2 and formate scavenger. The purified enzymes exhibited extremely high formate oxidation and CO2-reduction rates, and low Km values for formate. For the enzyme designated FDH-1, a specific formate oxidation rate of 700 U.mg-1 and a Km for formate of 0.04 mm were measured when benzyl viologen was used as an artificial electron acceptor. The enzyme designated FDH-2 oxidized formate with a specific activity of 2700 U.mg-1 and a Km of 0.01 mm for formate with benzyl viologen as electron acceptor. The specific CO2-reduction (to formate) rates measured for FDH 1 and FDH-2, using dithionite-reduced methyl viologen as the electron donor, were 900 U.mg-1 and 89 U.mg-1, respectively. From gel filtration and polyacrylamide gel electrophoresis it was concluded that FDH-1 is composed of three subunits (89 +/- 3, 56 +/- 2 and 19 +/- 1 kDa) and has a native molecular mass of approximately 350 kDa. FDH-2 appeared to be a heterodimer composed of a 92 +/- 3 kDa and a 33 +/- 2 kDa subunit. Both enzymes contained tungsten and selenium, while molybdenum was not detected. EPR spectroscopy suggested that FDH-1 contains at least four [2Fe-2S] clusters per molecule and additionally paramagnetically coupled [4Fe-4S] clusters. FDH-2 contains at least two [4Fe-4S] clusters per molecule. As both enzymes are produced under all growth conditions tested, but with differences in levels, expression may depend on unknown parameters. PMID- 12755704 TI - Ycf1p-dependent Hg(II) detoxification in Saccharomyces cerevisiae. AB - In Saccharomyces cerevisiae, disruption of the YCF1 gene increases the sensitivity of cell growth to mercury. Transformation of the resulting ycf1 null mutant with a plasmid harbouring YCF1 under the control of the GAL promoter largely restores the wild-type resistance to the metal ion. The protective effect of Ycf1p against the toxicity of mercury is especially pronounced when yeast cells are grown in rich medium or in minimal medium supplemented with glutathione. Secretory vesicles from S. cerevisiae cells overproducing Ycf1p are shown to exhibit ATP-dependent transport of bis(glutathionato)mercury. Moreover, using beta-galactosidase as a reporter protein, a relationship between mercury addition and the activity of the YCF1 promoter can be shown. Altogether, these observations indicate a defence mechanism involving an induction of the expression of Ycf1p and transport by this protein of mercury-glutathione adducts into the vacuole. Finally, possible coparticipation in mercury tolerance of other ABC proteins sharing close homology with Ycf1p was investigated. Gene disruption experiments enable us to conclude that neither Bpt1p, Yor1p, Ybt1p nor YHL035p plays a major role in the detoxification of mercury. PMID- 12755706 TI - NMR solution structure of calerythrin, an EF-hand calcium-binding protein from Saccharopolyspora erythraea. AB - The structure of calerythrin, a prokaryotic 20 kDa calcium-binding protein has been determined by solution NMR spectroscopy. Distance, dihedral angle, J coupling, secondary chemical shift, residual dipolar coupling and radius of gyration restraints reveal four EF-hand motifs arranged in a compact globular structure. A tight turn in the middle of the amino acid sequence brings the two halves, each comprising a pair of EF-hands, close together. The structural similarity between calerythrin and the eukaryotic sarcoplasmic calcium-binding proteins is notable. PMID- 12755705 TI - Trehalose influence on beta-lactoglobulin stability and hydration by time resolved fluorescence. AB - The stabilizing role of the disaccharide trehalose on beta-lactoglobulin (BLG) against its chemical denaturation both at native and acidic pH has been explored by means of time-resolved fluorescence of the probe acrylodan covalently bound to the unique free cysteine of BLG. The changes in acrylodan fluorescence lifetime with guanidinium chloride concentration reveal BLG sigmoidal denaturation profiles which depend upon the amount of trehalose in solution. When adding trehalose the transition midpoint shifts towards higher denaturant concentration. This effect has been measured by fitting the data with a two-state model whose parameters indicate that an almost 60% increase in the denaturation free energy is induced independently of trehalose concentrations and pH values. Fluorescence anisotropy measurements performed in the same conditions reveal that the internal dynamics are largely affected by the sugar, which makes the acrylodan environment more rigid, and by the denaturant that acts in the opposite way. The overall rotational diffusion of BLG suggests that trehalose affects the hydrodynamic properties of the solution in the proximity of the protein; tentative mechanisms are discussed. PMID- 12755707 TI - A novel type of family 19 chitinase from Aeromonas sp. No.10S-24. Cloning, sequence, expression, and the enzymatic properties. AB - A family 19 chitinase gene from Aeromonas sp. No.10S-24 was cloned, sequenced, and expressed in Escherichia coli. From the deduced amino acid sequence, the enzyme was found to possess two repeated N-terminal chitin-binding domains, which are separated by two proline-threonine rich linkers. The calculated molecular mass was 70 391 Da. The catalytic domain is homologous to those of plant family 19 chitinases by about 47%. The enzyme produced alpha-anomer by hydrolyzing beta 1,4-glycosidic linkage of the substrate, indicating that the enzyme catalyzes the hydrolysis through an inverting mechanism. When N-acetylglucosamine hexasaccharide [(GlcNAc)6] was hydrolyzed by the chitinase, the second glycosidic linkage from the nonreducing end was predominantly split producing (GlcNAc)2 and (GlcNAc)4. The evidence from this work suggested that the subsite structure of the enzyme was (-2)(-1)(+1)(+2)(+3)(+4), whereas most of plant family 19 chitinases have a subsite structure (-3)(-2)(-1)(+1)(+2)(+3). Thus, the Aeromonas enzyme was found to be a novel type of family 19 chitinase in its structural and functional properties. PMID- 12755709 TI - State of histone modification in the rat Ig-beta/growth hormone locus. AB - The state of acetylation in H3 and H4 histones and dimethylation in the H3 histone Lys4 residue were examined by chromatin immunoprecipitation (ChIP) at 11 targets in the rat Ig-beta/growth hormone locus. Marked enhancement of the acetylation of histones H3 and H4 and the dimethylation of H3 Lys4 was observed in the chromatin situated close to the promoter of an actively transcribed gene. Chromatin positioned near a cell-type-specific DNase I-hypersensitive site with enhancer activity had the same histone modifications as the active promoter. In one transcribed intron, chromatin with fewer histone modifications was found, and in another transcribed intron, chromatin with markedly enhanced modifications was found. In most cases, no appreciable difference in the acetylation of histones H3 and H4 was found at prominently enhanced targets. However, different acetylation levels of H3 and H4 were found at one target. The targets with enhanced dimethylation of the H3 Lys4 residue coincided with those with prominently enhanced acetylation of histones H3 and H4. PMID- 12755708 TI - A novel homocysteine-responsive gene, smap8, modulates mitogenesis in rat vascular smooth muscle cells. AB - We isolated the cDNA of a gene, designated smooth muscle-associated protein 8 (smap8), during a search for new genes expressed in human aortic smooth muscle cells. The full-length smap8 cDNA is 3241 bp long and contains an open reading frame of 1113 bp encoding an approximately 45 kDa soluble protein identical to NDRG4 protein. Smap8 mRNA was expressed predominantly in the brain and heart, and moderately in vascular smooth muscle cells. Expression of smap8 mRNA was induced within 3-12 h by treatment with 10 mm homocysteine in rat aortic smooth muscle cells (A10 cells). Expression of exogenous smap8 markedly reduced both the proliferation and migration rates of rat A10 cells, however, PDGF-induced proliferation was significantly enhanced in smap8-expressed cells compared with mock-transfected cells. To ascertain the involvement of smap8 in mitogenesis, we tested the effects of stimulation of smap8, MEK1/2 or ERK1/2, which is known as a proliferation relating intermediate, by various growth factors and cytokines. PDGF was the most prominent in promoting phosphorylation of the smap8 protein. PDGF-dependent phosphorylation of smap8 was induced prior to ERK1/2 activation, and was repressed by staurosporine, a general inhibitor of serine/threonine kinases. Furthermore, activation of both MEK1/2 and ERK1/2 was markedly enhanced in these cells. Smap8 might therefore regulate the potentiation of ERK1/2 signalling induced by PDGF treatment. Our results imply that smap8 is involved in the regulation of mitogenic signalling in vascular smooth muscle cells, possibly in response to a homocysteine-induced injury. PMID- 12755711 TI - Diversity and dynamics of microbial communities in soils from agro-ecosystems. AB - Soil microbial communities are integrally involved in biogeochemical cycles and their activities are crucial to the productivity of terrestrial ecosystems. Despite the importance of soil microorganisms, little is known about the distribution of microorganisms in the soil or the manner in which microbial community structure responds to changes in land management. We investigated the structure of microbial communities in the soil over two years in a series of replicated plots, that included, cultivated fields, fields abandoned from cultivation and fields with no history of cultivation. Microbial community structure was examined by monitoring the relative abundance of ribosomal RNA (rRNA) from seven of the most common bacterial groups in soil (the Alpha and Beta Proteobacteria, Actinobacteria, Cytophagales, Planctomycetes, Verrucomicrobia and the Acidobacteria) and the Eukarya. These data reveal that soil microbial communities are dynamic, capable of significant change at temporal scales relative to seasonal events. However, despite temporal change in microbial community structure, the rRNA relative abundance of particular microbial groups is affected by the local environment such that recognizable patterns of community structure exist in relation to field management. PMID- 12755712 TI - Bacterial community structure and function in a metal-working fluid. AB - The diversity of bacterial populations colonizing spatially and temporally separated samples of the same metal-working fluid (MWF) formulation was investigated. Analyses were performed with a view to improve strategies for bioaugmentation of waste MWF in bioreactor systems and prevention of in-use MWF biodeterioration in engineering workshops. Significantly, complementary phenotypic, genotypic and in situ methods revealed that the bacterial communities in operationally exhausted MWFs had low diversity and were similar in species composition from different locations and uses. Of the 179 bacterial isolates studied, only 11 genera and 15 species were identified using fatty acid methyl ester (FAME) analysis, with culture independent analyses by 16S rDNA denaturing gradient gel electrophoresis (DGGE) and fluorescent in situ hybridization being congruent with these FAME data. In order to gain some insight into functional role of detected populations, we correlated the MWF chemical composition and potential pollution load with bacterial abundance and community composition detected within samples. PMID- 12755713 TI - Extracellular phosphatase activity of natural plankton studied with ELF97 phosphate: fluorescence quantification and labelling kinetics. AB - ELF(R)97 phosphate (ELFP) is a phosphatase substrate which produces ELF(R)97 alcohol (ELFA), a fluorescent water-insoluble product, upon hydrolysis. We studied the kinetics of ELFA precipitation in freshwater samples at levels of total plankton and single phytoplankton cells, and tested the suitability of ELFP for measurement of surface-bound algal extracellular phosphatases. Samples from acidic Plesne Lake (pH approximately 5; high phosphatase activity) and eutrophic Rimov reservoir (pH approximately 7-10; moderate phosphatase activity) were incubated with ELFP for 5-300 min, fixed with HgCl2 and filtered through polycarbonate filters. Relative fluorescence of filter-retained ELFA precipitates was quantified with image analysis. Time-courses of ELFA formation exhibited lag periods followed by finite periods of linear increase. In Plesne Lake, lag-times were shorter (1-18 min) and rates of increase in ELFA fluorescence higher (by approximately 2 orders of magnitude) than in Rimov reservoir (lag-times 30-200 min). Similar patterns of ELFA formation kinetics were also observed in Plesne Lake samples in cuvette spectrofluorometer measurements (which failed in Rimov reservoir). Linear regression of seasonal data on rates of increase in ELFA fluorescence from image cytometry and spectrofluorometry (r2 = 0.65, n = 10) allowed for calibration of image cytometry in terms of amount of cell-associated ELFA. Preliminary measurements of extracellular phosphatase activities of several algae resulted in rates (10-2260 fmol cell-1 h-1) which are comparable to data reported in the literature for algal cultures. PMID- 12755714 TI - UV-acclimation responses in natural populations of cyanobacteria (Calothrix sp.). AB - Phenotypic acclimation to changing conditions is typically thought to be beneficial to organisms in the environment. UV radiation is an important parameter affecting photosynthetic organisms in natural environments. We measured the response of photosynthetic carbon fixation in populations of cyanobacteria inhabiting a hot spring following acclimation to different UV treatments. These two very closely related populations of cyanobacteria, differing in their content of the extracellular UV-screening pigment scytonemin, were acclimated in situ under natural solar irradiance modified by filters that excluded both UVA/B, only UVB or transmitted both UVA/B. Cells from each preacclimation treatment were subsequently assayed for photosynthetic performance under all UV conditions (incubation treatment) giving a two-factor experimental design for each population. No acclimation filter treatment effects were observed even after two months under different acclimation treatments. This suggests that UV photoacclimation does not occur in either of these populations, regardless of the presence of scytonemin. By contrast, cells showed significant UV-inhibition during 1 h incubations under full sun. The population with high levels of scytonemin usually had lower rates of photosynthetic carbon fixation than the scytonemin-lacking population. However, the degree of UV inhibition, especially UVA inhibition, was higher for the cells without scytonemin pigment. These results suggest that closely related natural cyanobacterial populations respond differently to natural irradiance conditions and may be adopting different strategies of UV tolerance. PMID- 12755715 TI - The synthesis of the UV-screening pigment, scytonemin, and photosynthetic performance in isolates from closely related natural populations of cyanobacteria (Calothrix sp.). AB - Two populations of the cyanobacterium Calothrix sp. found in Yellowstone thermal spring outflows differ greatly in their contents of scytonemin, a UV-screening pigment, and in their photosynthetic carbon assimilation rates. Clonal isolates from both populations were used to investigate these phenotypic differences. Identical partial 16S rDNA sequences ( approximately 900 bp) suggest a very close relationship between the two Calothrix populations and indicate that environmental differences may, in part, explain the field observations. The effects of native spring water on scytonemin synthesis and photosynthesis were tested during experiments using plated cells. Results show differences in the spring water environment were at least partly responsible for the differences in scytonemin content observed in the field. Furthermore, spring water effects on photosynthetic performance suggest adaptation in these strains to their spring of origin. Controlled experiments performed using cultures grown in artificial liquid medium showed no significant difference in photosynthetic carbon uptake between strains. However, significant differences were detected in their ability to synthesize scytonemin indicating genetic differences between populations. These findings suggest that both genetic and environmental differences are responsible for the naturally occurring variation in scytonemin content and photosynthetic ability in these two closely related populations. PMID- 12755716 TI - Archaeal diversity associated with in situ samplers deployed on hydrothermal vents on the East Pacific Rise (13 degrees N). AB - To evaluate possible compositional changes in archaeal communities at a deep-sea hydrothermal vent field scale, we examined five different samples obtained after deploying in situ collectors for different times on three spatially separated venting sulphide structures on the East Pacific Rise (13 degrees N). Direct cell counts and whole-cell hybridizations with fluorescently labelled 16S rRNA-based oligonucleotide probes revealed that the relative abundance of archaeal populations represented from 14 to 33% of the prokaryotic community. 16S rDNA sequence analysis of the archaeal clone libraries indicated that a large percentage of clones were closely related to known archaeal isolates recovered from similar habitats. Among the 24 different phylotypes identified, Thermococcales-related sequences were dominant in all the libraries that also included representative genera of orders Methanopyrales, Methanococcales, Archaeoglobales and Desulfurococcales. The presence of most of these phylogenetic groups was confirmed in enrichment cultures performed at temperatures from 60 to 90 degrees C. Additional sequences with no known cultivated relatives grouped with the Marine group I Crenarchaeota, Korarchaeota and Deep-sea Hydrothermal Vent Euryarchaeota (DHVE) within which a novel lineage was identified. Furthermore, the archaeal community composition was distinct from vent to vent within the same vent field and varied within short time scales. This study provides new insights into microbial diversity and distribution at deep-sea hydrothermal vents. PMID- 12755717 TI - Marinobacterium sp. strain DMS-S1 uses dimethyl sulphide as a sulphur source after light-dependent transformation by excreted flavins. AB - Marinobacterium sp. strain DMS-S1 is a unique marine bacterium that can use dimethyl sulphide (DMS) as a sulphur source only in the presence of light. High performance liquid chromatography (HPLC) analyses of the culture supernatant revealed that excreted factors, which could transform DMS to dimethyl sulphoxide (DMSO) under light, are FAD and riboflavin. In addition, FAD appeared to catalyse the photolysis of DMS to not only DMSO but also methanesulphonate (MSA), formate, formaldehyde and sulphate. As strain DMS-S1 can use sulphate and MSA as a sole sulphur source independently of light, the excretion of flavins appeared to support the growth on DMS under light. Furthermore, three out of 12 marine bacteria from IAM culture collection were found to be able to grow on DMS with the aid of photolysis by the flavins excreted. This is the first report that bacteria can use light to assimilate oceanic organic sulphur compounds outside the cells by excreting flavins as photosensitizers. PMID- 12755718 TI - Extraradical mycelium of the arbuscular mycorrhizal fungus Glomus lamellosum can take up, accumulate and translocate radiocaesium under root-organ culture conditions. AB - Radiocaesium enters the food chain when plants absorb it from soil, in a process that is strongly dependent on soil properties and plant and microbial species. Among the microbial species, arbuscular mycorrhizal (AM) fungi are obligate symbionts that colonize the root cortex of many plants and develop an extraradical mycelial (ERM) network that ramifies in the soil. Despite the well known involvement of this ERM network in mineral nutrition and uptake of some heavy metals, only limited data are available on its role in radiocaesium transport in plants. We used root-organ culture to demonstrate that the ERM of the AM fungus Glomus lamellosum can take up, possibly accumulate and unambiguously translocate radiocaesium from a 137Cs-labelled synthetic root-free compartment to a root compartment and within the roots. The accumulation of 137Cs by hyphae in the root-free compartment may be explained by sequestration in the hyphae or by a bottleneck effect resulting from a limited number of hyphae crossing the partition between the two compartments. Uptake and translocation resulted from the incorporation of 137Cs into the fungal hyphae, as no 137Cs was detected in mycorrhizal roots treated with formaldehyde. The importance of the translocation process was indicated by the correlation between 137Cs measured in the roots and the total hyphal length connecting the roots with the labelled compartment. 137Cs may be translocated via a tubular vacuolar system or by cytoplasmic streaming per se. PMID- 12755719 TI - Monitoring of alkane-degrading bacteria in a sea-water microcosm during crude oil degradation by polymerase chain reaction based on alkane-catabolic genes. AB - Behaviour of microbial populations responsible for degrading n-alkanes, a major component of crude oil, was monitored during crude oil degradation in a sea-water microcosm by both traditional colony culturing and molecular techniques. A DNA extraction method applicable to crude oil-amended sea-water samples was developed to obtain DNA applicable to most probable number (MPN) polymerase chain reaction (PCR). The population of alkane-degrading bacteria responsible for degradation of n-alkanes in a crude oil-amended microcosm altered, so that shorter alkanes were degraded first by alkane-degrading bacteria possessing alkane hydroxylase genes from group I (Kohno et al., 2002, Microb Environ 17: 114-121) and longer ones afterwards by those possessing alkane hydroxylase genes from group II. Thus, the degradation mechanism of the n-alkanes can be clarified during crude oil degradation. Application of the method of detecting different types of alkane catabolic genes, as shown in the present study, enabled bacterial groups preferring alkanes of either shorter or longer chain lengths to be enumerated selectively. PMID- 12755722 TI - Evidence-based diagnosis in patch testing. AB - Evidence-based medicine (EBM) is defined as the integration of the best research evidence with clinical expertise and patient values. Based on the principles of EBM, we can conclude that patch testing is cost-effective only if patients are selected on the basis of a clear-cut clinical suspicion of contact allergy and only if patients are tested with chemicals relevant to the problem (high pretest probability). Random patch testing (low pretest probability) should be discouraged. Proper pretest probability assessment can only be done in expert centres, because problem-based testing requires both a thorough knowledge of the patch-test procedure and knowledge about potential sensitizers in a specific environment. PMID- 12755720 TI - Phylogeny and distribution of nitrate-storing Beggiatoa spp. in coastal marine sediments. AB - Filamentous sulphide-oxidizing Beggiatoa spp. often occur in large numbers in the coastal seabed without forming visible mats on the sediment surface. We studied the diversity, population structure and the nitrate-storing capability of such bacteria in the Danish Limfjorden and the German Wadden Sea. Their distribution was compared to the vertical gradients of O2, NO3- and H2S as measured by microsensors. The main Beggiatoa spp. populations occurred in a 0.5-3 cm thick intermediate zone, below the depth of oxygen and nitrate penetration but above the zone of free sulphide. The Beggiatoa spp. filaments were found to store nitrate, presumably in liquid vacuoles up to a concentration of 370 mM NO3-, similar to the related large marine sulphur bacteria, Thioploca and Thiomargarita. The observations indicate that marine Beggiatoa spp. can live anaerobically and conserve energy by coupling sulphide oxidation with the reduction of nitrate to dinitrogen and/or ammonia. Calculations of the diffusive nitrate flux and the potential sulphide oxidation by Beggiatoa spp. show that the bacteria may play a critical role for the sulphur cycling and the nitrogen balance in these coastal environments. 16S rDNA sequence analysis shows a large diversity of these uncultured, nitrate-storing Beggiatoa spp. Smaller (9-17 micro m wide) and larger (33-40 micro m wide) Beggiatoa spp. represent novel phylogenetic clusters distinct from previously sequenced, large marine Beggiatoa spp. and Thioploca spp. Fluorescence in situ hybridization (FISH) of the natural Beggiatoa spp. populations showed that filament width is a conservative character of each phylogenetic species but a given filament width may represent multiple phylogenetic species in a mixed population. PMID- 12755723 TI - Nickel-elicited systemic contact dermatitis. AB - 20 patients with systemic contact dermatitis due to nickel are described. Of these patients, 15 were female and 5 were male. Their mean age was 24.8 years (16 51 years). All had experienced contact dermatitis in the umbilical area due to continual contact with metal belt-buckles or buttons. Then, with long- or short term aggravation of such periumbilical dermatitis, commonly in summer, lesions spread to other sites such as the side of the neck, the flexures of the extremities, etc. All patients showed a positive patch test to nickel sulphate (2.5% in petrolatum) and the dimethylglyoxime test demonstrated the presence of free nickel on metal buttons or belt-buckles. Punch biopsies performed in 7 patients showed subacute dermatitis. After avoidance of continual exposure to objects containing nickel and foods rich in nickel, as well as treatment with oral antihistamines and topical corticosteroids, all patients improved or cleared. It has been reported that nickel can cause systemic contact dermatitis by some internal systemic route, such as oral intake, transfusion, inhalation, implantation of metal medical devices, etc. In our patients, we found that continual local skin contact could also elicit systemic contact dermatitis. PMID- 12755724 TI - The composition of fine fragrances is changing. AB - High frequencies of contact allergy to fragrance ingredients have been reported in recent years. Developments in analytical chemistry have made it possible to measure exposure to well-known fragrance contact allergens. It has been shown that exposure is widespread in different types of products. The products with the highest concentrations of allergens have been shown to be prestige perfumes intended for women. This investigation explores the possible development in formulation of prestige perfumes, with regard to their content of the chemically defined ingredients of the diagnostic patch test material, the fragrance mix (FM). 10 fine fragrances were subjected to chemical analysis: 5 of these had been launched years ago (1921-1990) and 5 were the latest launches by the same companies, introduced 2 months to 4 years before purchase. The analysis revealed that the 5 old perfumes contained a mean of 5 of the 7 target allergens of the FM, while the new perfumes contained a mean of 2.8 of the allergens. The mean concentrations of the target allergens were 2.6 times higher in the old perfumes than in the new perfumes, range 2.2-337. It is concluded that the old perfumes, which are still popular products on the market, have a different composition from the new perfumes. This may be due to change in fashion or to an effort by the fragrance industry to focus on fragrance contact allergy, especially that to the FM ingredients. PMID- 12755725 TI - Anaphylaxis to polyvinylpyrrolidone after vaginal application of povidone-iodine. AB - A 59-year-old woman who had had several episodes of contact urticaria after hair treatment, developed anaphylaxis after vaginal application of povidone-iodine solution for disinfection. Prick tests showed wheal-and-flare responses to both povidone-iodine (0.1% aqueous) and polyvinylpyrrolidone (povidone, PVP) (0.001% aq.), but not to iodine or polyoxy-ethyrenenonylphenyl ether, both of which are also contained in povidone-iodine solution. We confirmed that basophils from her peripheral blood released considerable amounts of histamine on stimulation by PVPs. It appeared that both the shampoo and the permanent-wave solution contained polyvinylpyrrolidone N, N-dimethyl aminoethyl methacrylic acid copolymer diethyl sulphate solution and polyvinylpyrrolidone styrene-copolymer emulsion. Both these agents in the hair care products provoked an immediate skin response on prick testing. We speculate that sensitization to PVP had been established by these hair care products at a beauty parlor. She was recommended to avoid PVP containing products and remained free from symptoms thereafter. PMID- 12755727 TI - Optimizing the patch-test concentration of para-tertiary-butylcatechol: results of a prospective study with a dilution series. AB - Para-tertiary-butylcatechol (PTBC), which has long been patch tested internationally at 1% and 0.5% concentrations, was recently shown to induce patch test sensitization at the 1% patch-test concentration. In order to determine a safe patch-test concentration, we performed a prospective study with lowered patch-test concentrations. A dilution series of PTBC 0.25%, 0.1%, 0.01% and 0.001% (petrolatum, pet.) was tested on the upper back. Additionally, 0.25% PTBC was tested on the left upper arm to allow patients to carry out self-examination daily. Patch tests were read on D1-3, D7, D14 and D21 after patch-test application. Patients who were not able to return for all scheduled readings were telephoned and asked to report any reaction at the patch-test sites. 65 out of the 101 patients included completed the study. A positive patch-test reaction was observed in 4 patients during D1-D3, indicating previous sensitization. 1 patient had a doubtful reaction at D3 reading. Negative patch-test results were noted in 60 patients. None of the patients developed a positive patch-test reaction during the late readings (D7-D21). Thus, patch-test sensitization was not observed in any case. Para-tertiary-butylcatechol 0.25% pet. is recommended for patch testing internationally. PMID- 12755726 TI - Patch testing with carbamazepine and its main metabolite carbamazepine epoxide in cutaneous adverse drug reactions to carbamazepine. AB - The value of skin tests in the diagnosis of adverse drug reactions (ADRs) has been limited. Lack of knowledge as to the nature of drug allergens has contributed to these limitations. Several reports have addressed the roles of metabolites in cutaneous ADRs and skin testing. We evaluated the role of a carbamazepine (CBZ) metabolite on the results obtained from patch tests, using CBZ and its main metabolite 10, 11-epoxide of CBZ (CBZ-epoxide), on 13 patients with CBZ-induced drug eruptions and 39 controls with no CBZ-induced cutaneous ADRs. 10 of the 13 patients showed a positive reaction, and 2 of the 10 patients had a reaction to the CBZ-epoxide only and 1 to both CBZ and CBZ-epoxide. None of the 39 controls displayed any reactions to either CBZ or CBZ-epoxide. Patch testing of suspected drugs, as well as their available metabolites, would be helpful in improving the results. PMID- 12755728 TI - Spectrum of cross-photosensitization in 18 consecutive patients with contact photoallergy to ketoprofen: associated photoallergies to non-benzophenone containing molecules. AB - Contact photoallergy to ketoprofen gels has been widely reported, and cross sensitivity reactions with other compounds, such as tiaprofenic acid, fenofibrate and benzophenones, are well known. However, positive photopatch tests to other different non-benzophenone-related compounds have recently been observed. We report the results of photopatch testing in patients with contact photoallergy to ketoprofen and discuss the spectrum of cross-sensitization to ketoprofen. 18 consecutive patients with a history of photocontact dermatitis from ketoprofen were investigated. Patch and photopatch tests were performed. As expected, we observed positive photopatch tests to Ketum* gel and ketoprofen 2.5% in petrolatum in all patients (100%). However, it was remarkable to note positive photopatch tests to other unexpected and non-relevant allergens, including fentichlor (67%), tetrachlorosalicylanilide (28%), triclosan (17%), tribromsalan (11%) and bithionol (11%), with no clinical relevance. Interestingly, these agents belong to the family of halogenated salicylanilides and related compounds, which have been forbidden in Europe since the 1970s. Our results raise the question of hyper-photosusceptibility to non-relevant allergens induced by photosensitivity to ketoprofen. The mechanism may involve the high photoreactivity induced by the association of a benzene ring with an oxygen group. PMID- 12755729 TI - Methyldibromo glutaronitrile: clinical experience and exposure-based risk assessment. AB - In the year 2000, the level of methyldibromo glutaronitrile (MDGN) allergy in dermatology clinics in Europe exceeded the level of allergies to all other preservatives, with a prevalence of 3.5%. In the present study, cases of primary sensitization and elicitation to MDGN due to cosmetic products were collected over an 8-month period at the Department of Dermatology, Gentofte University Hospital. The aim was to identify the products related to hand eczema, assess exposure to MDGN in these products and relate the findings to results from a newly developed updated risk assessment model for contact allergy. Out of 24 patients with a positive patch test to MDGN, 17 patients with hand eczema were identified. In 11 of these patients, cosmetic products used in relation to the onset of the disease were shown to contain MDGN (65%). In 8 of these 11 cases, primary sensitization was probable, 5 due to hand/body lotions and 3 due to lotions and/or liquid hand soap. Chemical analysis of 12 products showed that lotions contained 149-390 ppm of MDGN, liquid hand soap 144-399 ppm, a rinsing cream 293 ppm and shampoos 78-79 ppm. The shampoo exposure was not of certain relevance to the eczema. Applying the newly developed updated risk assessment model showed that the concentrations of MDGN in lotions of 149-390 ppm exceeded the calculated maximum acceptable exposure level for MDGN, which would be expected to lead to sensitization in consumers using such products, as seen in the current study. The present cases and updated exposure-based risk assessment process add to the evidence and need for re-defining safe-use concentrations of MDGN in cosmetic products. PMID- 12755730 TI - Airborne allergic contact dermatitis from 3-iodo-2-propynyl-butylcarbamate at a paint factory. AB - 3-Iodo-2-propynyl-butylcarbamate (IPBC) is a fungicide used in both industrial products and cosmetics. We report the first case of allergic contact dermatitis from airborne exposure to this preservative. A 34-year-old female production worker at a paint factory developed dermatitis on air-exposed skin areas. Patch testing showed a ++ reaction to the preservative IPBC 0.01% in petrolatum. The compound was used as a preservative in wood treatment products manufactured at her work place. Based on animal studies, IPBC is considered safe as a cosmetic preservative. However, widespread use of the chemical might lead to increasing levels of contact allergy, and therefore, close monitoring of IPBC is recommended. PMID- 12755731 TI - Routine patch testing with frullanolide mix: an European Environmental and Contact Dermatitis Research Group multicentre study. AB - Contact sensitivity to plants containing 1 or more sesquiterpene lactones (SLs) is difficult to diagnose. The mixture of SLs (SL mix) has been shown to detect only about 60% of sensitized individuals. In order to improve the diagnosis of sensitization to plants containing SLs, we have tested a mixture of frullanolides contained in Frullania dilatata and Frullania tamarisci at 3 different concentrations (0.01%, 0.033% and 0.1% in petrolatum). 8605 consecutive eczema patients in 1 North American and 15 European dermatology departments were tested with this mix, and 0.35% of positive cases to the different concentrations were found. Routine use of this mix permitted detection of only a small percentage of extra cases and did not improve the SL mix score. The frullanolide mix should therefore be restricted to investigations in particular geographical zones and/or in particular occupations. PMID- 12755732 TI - Contact sensitivity to calcium hypochlorite. AB - Eusol (an abbreviation for Edinburgh University solution), consisting of aqueous boric acid and calcium hypochlorite, is widely used in the management of open wounds left to heal by secondary intention (1). PMID- 12755733 TI - Drug intolerance reaction to insulin therapy caused by metacresol. PMID- 12755734 TI - Acute irritation followed by primary sensitization to 2-cyclohexen-1-one in a chemistry student. PMID- 12755735 TI - Glucosides as unexpected allergens in cosmetics. PMID- 12755736 TI - Immediate-type hypersensitivity and allergic contact dermatitis due to para phenylenediamine in hair dye. PMID- 12755737 TI - Delayed hypersensitivity reaction to the non-ionic X-ray contrast medium Visipaque (iodixanol). PMID- 12755738 TI - Allergic contact dermatitis due to DL-alpha-tocopheryl nicotinate. PMID- 12755739 TI - 'Buffalo-hump' dermatitis: a hat trick of antiretroviral side-effects. PMID- 12755740 TI - Clindamycin-associated acute generalized exanthematous pustulosis. PMID- 12755741 TI - Allergic contact dermatitis from formaldehyde with initially negative repeated open application test. PMID- 12755742 TI - Occupational airborne allergic contact dermatitis from mesna. PMID- 12755743 TI - Co-sensitivity between cobalt and other transition metals. PMID- 12755744 TI - Unusual widespread vesicular eruption related to dental composite resin sensitization. PMID- 12755745 TI - Trends in allergic contact dermatitis and preventive measures among cement workers (1985-1999). PMID- 12755746 TI - Occupational protein contact dermatitis due to Calliphora vomitoria larvae (maggots) bred as fishing bait. PMID- 12755747 TI - Allergic contact dermatitis from pentylene glycol in an emollient cream, with possible co-sensitization to resveratrol. PMID- 12755749 TI - Scaffolds and biomaterials for tissue engineering: a review of clinical applications. AB - Tissue engineering is a multidisciplinary area of research aimed at regeneration of tissues and restoration of organ function. This is achieved through implantation of cells/tissues grown outside the body or by stimulating cells to grow into an implanted matrix. In this short review, we discuss the use of biomaterials, in the form of scaffolds, for tissue engineering and review clinical applications to otorhinolaryngology-head and neck surgery. PMID- 12755750 TI - The AAO-HNS Committee on Hearing and Equilibrium guidelines for the diagnosis and evaluation of therapy in Meniere's disease: have they been applied in the published literature of the last decade? AB - To assess how effectively the American Academy of Otolaryngology-Head and Neck Surgery Committee on Hearing and Equilibrium (AAO-HNS CHE) guidelines for the diagnosis and evaluation of therapy in Meniere's disease have been applied in the last 11 years of published literature. This was a MedLine-based review. Some 79.7% of papers attempted to use the AAO-HNS CHE guidelines. However, only 50% of these publications managed to use the AAO-HNS CHE criteria in the diagnosis and evaluation of therapy correctly. In order to advance our understanding of this condition, improved application of the AAO-HNS CHE guidelines by authors and editors alike is required in the reporting of results of the therapy of Meniere's disease. PMID- 12755751 TI - Does asbestos cause laryngeal cancer? AB - Asbestos is a known carcinogen. Its role in the aetiology of laryngeal cancer has been investigated and debated for the last three decades. Here, we consider much of the published evidence from post-mortem studies, cross-sectional, case-control and cohort studies. We feel that the weight of evidence does not support a causal association for asbestos with laryngeal cancer. The 'positive studies' raise important questions regarding an association; the opportunity may present itself to answer some of these as the incidence of asbestos-related malignancy is increasing in the UK and Europe. PMID- 12755752 TI - Could the presence of a Carhart notch predict the presence of glue at myringotomy? AB - The Carhart notch is a dip in the bone conduction at 2000 Hz without a corresponding dip in the air conduction. The main objective of this study was to establish how reliable is the presence of the Carhart notch in a preoperative audiogram in predicting the presence of glue at myringotomy. A prospective study has been carried out in 50 children presenting with glue ear to find out the association between the Carhart notch and the presence of glue at myringotomy. Children were seen before the operation and an audiogram and tympanogram were carried out. Myringotomy was carried out in 95 ears and the presence or absence of glue was recorded. The significance of the air-bone gap and the type of tympanogram in predicting a middle ear effusion were also examined. The audiograms of 37 ears showed a Carhart notch; of these, 36 ears were noted to have glue ear. The correlation between the presence of a Carhart notch in the preoperative audiogram and the presence of glue at myringotomy was found to be significant (P < 0.001) (chi-square test). Using the decision rule that the presence of a Carhart notch predicts the presence of glue, the following operating characteristics were determined: correct, 83 ears (87.4%); false positive, one ear (1.1%); false negative, 11 ears (11.6%). The presence of a Carhart notch was found to be a strong predictor of the presence of glue at myringotomy. PMID- 12755753 TI - Wegener's granulomatosis: an analysis of 33 patients seen over a 10-year period. AB - This retrospective study examined 33 patients with Wegener's granuloma seen between 1990 and 1999 in the Ayrshire and Arran region of Scotland. There was an estimated annual incidence of 10/million/year. The diagnosis in this series was based on the presence of one or more of the following: a positive histology, a positive c-ANCA or a typical clinical course. Twenty-eight patients were diagnosed based on either a positive histology and/or c-ANCA, whereas the remaining five were diagnosed based on the clinical course. c-ANCA was positive in 79.3% but correlated poorly with disease activity. Nasal biopsies were positive in 40%, whereas 94% of renal biopsies were positive, thereby making nasal biopsies unreliable in the diagnosis. Significantly elevated levels of erythrocyte sedimentation rate (ESR) averaging 77 mm/h were found in 32 patients at diagnosis. This showed fluctuation with disease activity. Thirteen patients died, 12 within 5 years. The best prognostic indicator statistically was age. PMID- 12755754 TI - Objective assessment of auditory thresholds in noise-induced hearing loss using steady-state evoked potentials. AB - The purpose of this study is to evaluate whether steady-state evoked potential (SSEP) can be used for objective estimation of auditory thresholds in patients with noise-induced hearing loss (NIHL). Eleven subjects (22 ears) with a characteristic audiometric notch between 3000 and 6000 Hz participated in this study. Both pure-tone thresholds and SSEP thresholds were obtained for each ear of all subjects. The correlation of SSEP thresholds and pure-tone thresholds was assessed. The results show that SSEP thresholds predicted pure-tone thresholds with correlation coefficients (r) of 0.86, 0.92, 0.94 and 0.95 at 500, 1000, 2000 and 4000 Hz respectively. Typically, the SSEP thresholds overestimate the pure tone thresholds by 10-20 dB, but they closely reflect the configuration of the audiogram. The strength of the relationship between SSEP and pure-tone thresholds increased with increasing frequency and increasing degree of hearing loss. In conclusion, SSEP can be used as a reliable and objective tool to assess auditory thresholds in patients with noise-induced hearing loss with high-frequency dips. PMID- 12755755 TI - Is it necessary to screen for hearing loss in the paediatric population with osteogenesis imperfecta? AB - The aim of the study was to assess the necessity of a screening service to detect early hearing loss in the paediatric population with osteogenesis imperfecta. Twenty-two children were assessed over a 5-year period. Five children (22.7%) had normal hearing. Fourteen (63.6%) had conductive hearing loss, with 12 children in this group having otitis media with effusion (OME); all had resolution of hearing loss with appropriate therapy. Two children had persistent conductive losses unrelated to OME. Three children (13.6%) had sensorineural hearing loss, with one being detected at the age of 1 year. Existing evidence suggests that hearing loss associated with osteogenesis imperfecta has its onset in the second to third decade of life. Contrary to this, hearing loss was detected in 77.3% (17) of this population with a median and mean age of 9 years. This study would suggest that routine screening is worthwhile in children with osteogenesis imperfecta. PMID- 12755756 TI - Otological trauma resulting from the Soho Nail Bomb in London, April 1999. AB - We report the otological effects of the April 1999 Soho Nail Bomb on 17 patients. Twenty-one (62%) tympanic membranes were perforated (pars tensa only); 78% closed spontaneously within 6 months. The mean size of the perforation in the tympanic membrane nearer to the blast was significantly larger than the opposite side [33% +/- 8.3 (mean +/- SD) and 13% +/- 4.1 respectively; P = 0.02]. All patients reported hearing losses that were mixed conductive and sensorineural but mainly high-frequency sensorineural (4, 6 and 8 kHz, pure tone average 42.3 dB +/- 20.5). The sensorineural hearing loss correlated inversely with the distance from the explosion but not with the size of perforation. There was no significant difference in the hearing loss between the ear facing the blast and the opposite ear. Fifteen patients (88%) had temporary tinnitus. No patient complained of any vestibular symptoms. The otological effects of a nail bomb in an enclosed space have not been previously reported. Furthermore, an inverse correlation between hearing loss and distance from the explosion and a significant difference in perforation size facing the blast, compared with the opposite side, are also presented for the first time. The high spontaneous closure rate of perforations and minimal ongoing disability from sensorineural losses favour conservative management in most cases. PMID- 12755757 TI - The place of endoscopic sinus surgery in the treatment of paranasal sinus mucocoeles. AB - Mucocoeles of the paranasal sinuses often enlarge slowly resulting in local bone erosion with subsequent extension into the adjacent orbit or intracranial space. We have reviewed the management of 59 patients with 68 mucocoeles. The variety of presentations is examined and discussed. Although diplopia has previously been stated as a very common finding we have found this not to be the case and believe that it may be that previous reports originate from ophthalmology centres. Forty four patients underwent endoscopic surgical management of their mucocoeles, nine had a combined external and endoscopic procedure and 14 patients had an external approach. One patient needed no surgery. The mean follow-up period was 6 years 3 months. There was the lowest number of recurrences in the endoscopic surgical group. We emphasize the importance of long-term follow-up. PMID- 12755758 TI - Delayed facial palsy following laser stapedectomy: in vitro study of facial nerve temperature. AB - Delayed facial palsy following conventional stapes surgery is a rare event, but this complication appears to be more common when a laser is used. We have investigated the temperature in the facial canal during stapes surgery using a KTP laser or a microdrill in preserved human temporal bones. Thermocouples were placed in the facial canal and under the foot plate. The results show maximum rises in temperature of between 1.4 degrees C and 15.2 degrees C in the facial canal during laser surgery (mean 6.1 degrees C, SD 4.5 degrees C), but only between 0.45 degrees C and 2 degrees C during procedures in which a microdrill was used (mean 0.9 degrees C, SD 0.9 degrees C) (P = < 0.009). In addition, the facial nerve undergoes repeated heating and cooling cycles during the laser surgery. We conclude that heating of the facial nerve during laser surgery causes oedema, which in turn leads to compression of the nerve within its bony canal. PMID- 12755759 TI - The modified endoscopic Lothrop procedure in the treatment of complicated chronic frontal sinusitis. AB - The purpose of this study was to evaluate the modified endoscopic Lothrop procedure in the management of complicated frontal sinus disease which has breached the confines of the sinus walls and extended into the cranial cavity or orbit. Fourteen patients with radiological evidence of 17 complications of frontal sinus disease presented over a 23-month period. CT scan and MRI scans revealed the presence of posterior table erosion and extension of the frontal sinus disease into the anterior cranial fossa in 10 patients. In addition, seven patients had intraorbital complications, with three patients having both intracranial and orbital complications. All patients underwent a modified endoscopic Lothrop procedure as part of the management of the complication. In addition, one patient required an orbital abscess drainage and repair of an encephalocele, with a second patient requiring drainage of an orbital subperiosteal abscess. At follow-up, all patients were asymptomatic and had patent frontal sinus ostia. Follow-up ranged from 8 months to 38 months with a median of 25 months. Three patients required a revision of their frontal ostium. Two patients had allergic fungal sinusitis with aggressive polyp recurrence and ostial re-stenosis while one patient developed recurrent orbital infections from a retained frontal sinus cell. Currently, all have patent ostia, with an average size of 14.6 x 11 mm. The modified endoscopic Lothrop procedure is an effective form of treatment in the management of complicated frontal sinus disease. The results are comparable to those achieved with other surgical approaches such as the osteoplastic flap with obliteration. PMID- 12755760 TI - Lymph node metastases in the lower neck. AB - Current knowledge suggests that lymph node metastases in the lower neck (supraclavicular fossa and posterior triangle) are associated with a poor survival. Very little systematic work has been published on this subject. This was a retrospective study carried out on a database where all patients were entered in a prospective manner over a 35-year period using a standard pro-forma. Data on 168 patients presenting with a lower neck node metastasis were retrieved. The main outcome measures were: association between variables and tumour-specific survival. Data were displayed in contingency tables and analysed by chi-square and categorical modelling. Recurrence and survival were plotted in a cause specific manner using the Kaplan Meir method. Differences in curves were analysed using the log rank test. Multivariate analysis was carried out using Cox's proportional hazard model. The only association was between site and node level and histology. Head and neck tumours were associated with squamous histology (P = 0.0004) and supraclavicular nodes (P = 0.0047). Survival time was not significantly different when lower-neck lymph node metastasis from the head and neck was compared to non-head and neck metastasis: 5-year survival 30% and 10% respectively (P = 0.1363). Survival with posterior triangle metastases was significantly better than supraclavicular metastases (P = 0. 0059), confirmed on multivariate analysis. Laterality of metastasis had no effect on survival (P < 0.0001). There was no significant difference in survival between squamous and non squamous metastases on Cox regression (P = not significant). There were 85 head and neck primaries including lymphomas, 53 infraclavicular primaries and 30 unknown primaries. There were 73 squamous cell carcinomas, 27 adenocarcinomas, 34 lymphomas, 28 undifferentiated tumours and six other tumours. Nearly half the primary tumours were below the clavicle. Survival was unaffected by laterality, primary site or histology, but was better for posterior triangle nodes. PMID- 12755761 TI - Hearing loss in adult Refsum's disease. AB - Refsum's disease is characterized by defective peroxisomal alpha oxidation of phytanic acid, with clinical features that include retinitis pigmentosa, polyneuropathy, anosmia and hearing loss. Although hearing loss in Refsum's disease is common, there are few detailed assessments of the site of the abnormality. We examined the audiometric findings in patients with biochemically diagnosed Refsum's disease in order to assess the site of origin of the hearing loss. We found hearing loss, ranging from mild, predominantly high frequency to moderate degree, in seven out of nine patients with biochemically diagnosed adult Refsum's disease. In addition, we found evidence to suggest subtle auditory nerve involvement in six out of the seven patients with hearing loss and in one out of the two patients with a normal pure tone audiogram, on the basis of the ABR test results. We conclude that patients with Refsum's disease who report hearing difficulties should have full audiometric investigations in order to provide appropriate audiological rehabilitation. PMID- 12755762 TI - Increasing incidence of advanced stage head and neck tumours. AB - The aim of this study was to determine whether the incidence of advanced stage (T4) head and neck tumours has increased. We analysed retrospectively 3178 patients diagnosed with oral, pharyngeal or laryngeal cancer in the period 1980 2000 at the University Medical Center Utrecht (UMCU), The Netherlands. There was a statistically significant increase in the proportion of T4 head and neck tumours compared with non-T4 tumours over the period 1980-2000. Linear regression analysis estimated an increase of 0.9% every year. The observed increase in T4 tumours at UMCU shows up in figures from the Netherlands Regional Cancer Registry (IKMN) and the National Cancer Registration (NCR). Although these bodies report for fewer years, this finding refutes the possibility of selected referral to the University Medical Center Utrecht. In conclusion, the number of head and neck cancer patients presenting with an advanced stage carcinoma (T4) has increased over a period of 21 years. PMID- 12755763 TI - Comparison of histology between recurrent tonsillitis and tonsillar hypertrophy. AB - To elucidate the aetiology of tonsillar hypertrophy and recurrent tonsillitis, it is important to determine whether a difference exists between these two conditions in microanatomical architecture. The aim of this study was to investigate the difference in follicle size and numbers in tonsils for patients with tonsillar hypertrophy and recurrent tonsillitis using an image analysis method. Our results showed that there was no significant difference in the mean follicle numbers per counting field (40x magnification) between recurrent tonsillitis (4.5 +/- 2.1) and tonsillar hypertrophy (3.8 +/- 0.5). However, the mean follicle area in the tonsillar hypertrophy group (0.23 +/- 0.02 mm2) is significantly (P < 0.01) larger than in the recurrent tonsillitis group (0.15 +/- 0.02 mm2). Our study demonstrates that tonsillar hypertrophy is characterized histologically by an enlargement of follicles compared with chronic tonsillitis, indicating a hyperplastic condition of lymphoid cells in the germinal centres. It may also explain the difference in aetiology and/or immune defence mechanism underlying these two conditions. PMID- 12755764 TI - ENT presentations in children with HIV infection. AB - Illnesses of the ear, nose and throat (ENT) are common in children with human immunodeficiency virus (HIV) infection. We reviewed the case files of 107 HIV seropositive children in the paediatric HIV unit at St Mary's Hospital. The prevalence, age of onset and type of ENT disease were reviewed. We also determined sex distribution, maternal country of origin and mode of transmission of HIV. Fifty per cent of the HIV children had ENT illnesses. Fifty-five per cent of the children presented with their first ENT symptom before age 3 years with 98% of the children having ENT manifestations by age 9 years. The commonest ENT diseases were cervical lymphadenopathy (70%), otitis media (46%), oral candidiasis (35%) and adenotonsillar disease (31%). HIV transmission was vertical in 90%. Maternal country of origin was Africa in 70% and the UK in 13%. Compared with previous studies, the proportion of HIV children with ENT problems appears to have decreased. Although our figures report a similar ENT symptom profile, the age at onset of these symptoms has increased. PMID- 12755765 TI - Endovascular treatment of epistaxis in patients irradiated for nasopharyngeal carcinoma. AB - Posterior epistaxis commonly occurs after radiotherapy for nasopharyngeal carcinoma and is difficult to treat. Endovascular treatment was carried out on 11 previously irradiated patients who presented with intractable epistaxis over 4 years. We review their outcomes. All had significant angiographic abnormalities. Ten patients underwent intra-arterial embolization and, in one patient, balloon occlusion of an arterial pseudo-aneurysm was performed. Two patients (18%) suffered permanent neurological deficits. Two (18%) required further embolization in the first 2 days and one (9%) after the first month. One patient (9%) is still alive 4 years later. Only three (27%) died from haemorrhage. The mean survival duration after treatment was 225 days. Post-radiation anatomical changes often make conventional treatment of epistaxis difficult. Endovascular treatment maps out aberrant vascular anatomy and identifies the offending vessel. Our results show that the survival benefit of this treatment outweighs the risks, as such haemorrhage is often fatal. PMID- 12755766 TI - Assessment of surgical competence in parotid surgery using a CUSUM assessment tool. AB - The introduction of professional revalidation and clinical governance imposes an increasing requirement for clinicians to provide evidence of competence. This paper introduces a CUSUM (CUmulative SUM) mathematical tool for determining trend and applies the technique to 84 consecutive parotidectomies (superficial, partial or total/near total) performed by one surgeon (J.S.). Using this operation, the paper illustrates how attainment and maintenance of competence in the field of otolaryngological surgery may be demonstrated at differing levels of rigour. The role of the CUSUM in assessing a trainee's surgical progress is also explored. PMID- 12755767 TI - Hearing preservation and facial nerve function in vestibular schwannoma surgery. AB - The objective of this study was the assessment of hearing preservation in vestibular schwannoma surgery. This study reports a prospective cohort of 40 patients with clinically useful hearing from a consecutive series of 191 patients with unilateral vestibular schwannoma. The patients were managed in a tertiary centre by a combined team of Neurosurgeon, Otologist and Neuro-monitoring Scientist via a neuro-oto-surgical-retrosigmoid approach. Pure tone speech audiometry was conducted preoperatively and 6 months to 9 years following surgery. Using the AA0-HNS classification, useful hearing, i.e. grades A, B and C, was preserved in 47.5% of patients. Thirty-eight per cent achieved grade A or B. Using appropriate surgical and monitoring techniques, it is possible to preserve useful hearing in approximately 50% of patients following removal of a vestibular schwannoma via the retro-sigmoid approach. PMID- 12755768 TI - Palatal surgery for snoring: objective long-term evaluation. AB - Snoring surgery has mainly been assessed in the literature using subjective outcome measures with some evidence to indicate that the benefits of surgery are short lived. There is remarkably little published on the objective outcome of snoring surgery. We reported previously short-term (2-11 months) objective results on 38 patients who had palatal surgery. All the patients had preoperative and postoperative objective assessment of their snoring loudness and duration in the home. The current study presents the long-term objective results of 24 patients from the same cohort. All patients who were included in our study had palatal snoring confirmed on sleep nasendoscopy. and none of them had significant obstructive sleep apnoea. The median follow-up period was 45 months (range 29-56 months). Overall, the mean snore index (SI) at long-term follow-up was significantly less than the preoperative SI (P < 0.0001). There was no significant difference between the short-term and the long-term results regarding the fall in the SI. This study is the first to use postoperative objective assessment in the home and to demonstrate that snoring reduction is maintained for several years after snoring surgery. PMID- 12755769 TI - Marginally excised parotid pleomorphic salivary adenomas: risk factors for recurrence and management. A 12.5-year mean follow-up study of histologically marginal excisions. AB - The aim of the study was to identify recurrence risk factors in surgically excised parotid pleomorphic salivary adenomas. We reviewed the case histories and histological findings for all cases of marginal or inadequate excision of pleomorphic salivary adenomas at the Royal Hallamshire Hospital, Sheffield, UK, between 1980 and 1995. A total of 83 cases with complete records were identified, with a mean follow-up period of 12.5 years. The histological slides were reviewed in each case. The overall recurrence rate was 6.0%. Where tumour was present at the margin, recurrence occurred in 17.6% of cases. However, cases conventionally regarded as marginally excised and likely to recur (tissue margin < 1 mm) showed recurrence in only 1.8% of cases. Intraoperative capsular rupture, microscopic capsular invasion by tumour and several other surgical factors were not predictive of recurrence. Adequate excision of pleomorphic salivary adenomas, in the sense of minimal recurrence risk, does not require more than a fraction of a millimetre of surrounding tissue. Only pleomorphic salivary adenomas with tumour actually at the excision margin require prolonged follow-up or consideration of radiotherapy. Provided that the tumour can be removed intact, the surgical approach for pleomorphic salivary adenomas should be guided by the need to preserve vital structures rather than by an attempt to remove a cuff of normal tissue with the tumour. PMID- 12755770 TI - The effect of combined spinal-epidural (CSE) anaesthesia and size of spinal needle on postoperative hearing loss after elective caesarean section. AB - The exact aetiology of vestibulocochlear dysfunction after spinal anaesthesia is unknown. Low-frequency hearing loss occurs after spinal anaesthesia. The aim of this study was to investigate the effects of combined spinal-epidural (CSE) anaesthesia and size of spinal needle on vestibulocochlear dysfunction, using pure tone audiometry performed pre- and on the first and the second day postoperatively. Forty-five patients who were to undergo elective caesarean section were evaluated. In group I, CSE anaesthesia (18 G Tuohy, 25 G Whitacre pencil-point-design spinal needles) was performed in 15 patients. In group II, spinal anaesthesia was performed in 15 patients with 25 G Whitacre pencil-point design spinal needles and, in group III, spinal anaesthesia was performed in 15 patients with 22 G Whitacre pencil-point-design spinal needles. In the pre- and on the first and the second day postoperatively, the pure tone audiogram was performed in the audiology laboratory of our hospital, using a calibrated Kamplex Diagnostic Audiometer AC 40 in a noise-free room. When the CSE anaesthesia group and 22 G spinal group were compared for change in hearing between the pre- and postoperative periods, a statistically significant difference was observed at R right ear 125 Hz (P < 0.025) and at L-left ear 125 Hz (P < 0.023), and at L-left ear 1000 Hz (P < 0.036) and at R-right ear 1500 Hz (P < 0.006), and at L-left ear 1500 Hz (P < 0.022). At other frequencies, the difference was insignificant. When the CSE anaesthesia group and 25 G spinal group were compared for change in hearing between the pre- and postoperative periods, no statistically significant difference was detected at any frequency tested. When 22 G spinal group and 25 G spinal group were compared for change in hearing between the pre- and postoperative periods, there was some hearing loss at low frequency, although this difference did not reach statistical significance. The positive correlation of low-frequency hearing loss and increased pressure in the epidural space (which decrease the risk of cerebrospinal fluid leakage through the dura) suggests that cerebrospinal fluid leakage via the spinal puncture hole is not the only factor involved. Perioperative fluid replacement alone may not prevent hearing loss but CSF loss through the dural puncture site should also be prevented. PMID- 12755771 TI - Endoscopic bipolar diathermy in the management of epistaxis: an effective and cost-efficient treatment. AB - Epistaxis is one of the most frequently managed otorhinolaryngological emergencies. This prospective study over a 4-month period involved 38 adult patients presenting with epistaxis who underwent endoscopic bipolar diathermy under local anaesthesia. Thirty-four (89%) of the 38 adults were successfully treated and 28 (74%) of the patients did not require admission. Based on a simple cost-benefit analysis of savings made by avoiding admission, successful immediate treatment of these 28 patients led to a potential saving of at least pound 6804.00. We conclude endoscopic bipolar diathermy under local anaesthesia is an effective, safe and cost-efficient modality of treatment in the management of adult epistaxis. PMID- 12755772 TI - Management of suspected foreign body aspiration in children. PMID- 12755774 TI - De novo alveolar bone formation adjacent to endosseous implants. AB - OBJECTIVE: To describe a model for the investigation of different phases of wound healing that are involved in the process resulting in osseointegration. MATERIAL AND METHODS: The implants used for the study of early healing had a geometry that corresponded to that of a solid screw implant with an SLA surface configuration. A circumferential trough had been prepared within the thread region (intra osseous portion) that established a geometrically well-defined wound compartment. Twenty Labrador dogs received 160 experimental devices totally to allow the evaluation of healing between 2 h and 12 weeks. Both ground sections and decalcified sections were prepared from different implant sites. RESULTS: The experimental chamber used appeared to be conducive for the study of early phases of bone formation. The ground sections provided an overview of the various phases of soft and hard tissue formation, while the decalcified, thin sections enabled a more detailed study of events involved in bone tissue modeling and remodeling. The initially empty wound chamber became occupied with a coagulum and a granulation tissue that was replaced by a provisional matrix. The process of bone formation started already during the first week. The newly formed bone present at the lateral border of the cut bony bed appeared to be continuous with the parent bone, but woven bone was also found on the SLA surface at a distance from the parent bone. This primary bone that included trabeculae of woven bone was replaced by parallel-fibered and/or lamellar bone and marrow. Between 1 and 2 weeks, the bone tissue immediately lateral to the pitch region, responsible for primary mechanical stability of the device, became resorbed and replaced with newly formed viable bone. Despite this temporary loss of hard tissue contact, the implants remained clinically stable at all times. CONCLUSION: Osseointegration represents a dynamic process both during its establishment and its maintenance. In the establishment phase, there is a delicate interplay between bone resorption in contact regions (between the titanium body and mineralized bone) and bone formation in 'contact- free' areas. During the maintenance phase, osseointegration is secured through continuous remodeling and adaptation to function. PMID- 12755775 TI - Enzymatic degradation of collagen-guided tissue regeneration membranes by periodontal bacteria. AB - Bacterial infection in the vicinity of guided tissue regeneration barrier membranes was shown to have a negative effect on the clinical outcomes of this increasingly used technique. Several oral and specifically periodontal bacteria were shown to adhere to such membranes in vivo and in vitro with a higher affinity to membranes constructed from collagen. The present study examined the role of periodontal bacteria and their enzymes in the degradation of commercially used collagen membranes. Degradation of two collagen membranes [Biomend (Calcitek, Colla-Tec Inc., Plainsboro, NJ) and Bio-Gide (Geistlich Biomaterials, Wolhousen, Switzerland)] labeled by fluorescein isothiocyanate was examined by measuring soluble fluorescence. Porphyromonas gingivalis, Treponema denticola and Actinobacillus actinomycetemcomitans and their enzymes were evaluated. Collagenase from Clostridium hystolyticum was used as a positive control. While whole cells of P. gingivalis were able to degrade both types of membranes, T. denticola could degrade Bio-Gide membranes only and A. actinomycetemcomitans whole cells could degrade none of the membranes. Fractionation of P. gingivalis cells revealed that cell membrane associated proteases were responsible for the degradation of the two collagen membranes. In T. denticola, the purified major phenylalanine protease was found to be responsible for the degradation of Bio Gide membranes. These results suggest that proteolytic bacterial enzymes may take part in the degradation of collagen barrier membranes used for guided tissue regeneration. PMID- 12755776 TI - Effects of hydroxyapatite-coated and commercially pure titanium oral implant surfaces on compound nerve action potentials. AB - The aim of this study was to investigate the effects of hydroxyapatite-coated and commercially pure titanium oral implants on nerve conduction. Isolated rat sciatic nerves were placed between two suction electrodes in a bath containing a tyrode solution. The implants were brought into intimate contact with the nerves and evoked compound action potentials (cAPs) were recorded before and after contact with the implants. The commercially pure titanium implants did not cause any change in cAPs. A gradual reduction in cAPs was observed for hydroxyapatite coated implants. However, this reduction was < 50% after an application time of 120 min. Recovery of the cAPs in this group was recorded after approximately 60 min. We conclude that, although intimate contact with hydroxyapatite-coated implants leads to a reduction in cAPs in nerves in vitro, neither this surface nor a commercially pure titanium surface leads to irreversible neurotoxic effects. PMID- 12755777 TI - In-patient comparison of immediately loaded and non-loaded implants within 6 months. AB - According to the Branemark protocol, a stress-free healing period is one of the most emphasised requirements for implant integration. Recent studies have encouraged a progressive shortening of the healing period and immediate loading has been proposed for the edentulous mandible. This prospective study evaluated the clinical outcomes of 14 immediately loaded FRIALIT-2(R) implants compared with 28 non-loaded controls in an in-patient study. The results were based on clinical stability and on changes of bone level from implant placement to abutment connection 6 months after insertion. In the course of our investigation, seven patients with edentulous mandibles have been treated with 43 implants following an immediate-loading protocol. Six FRIALIT-2(R) implants were placed in the interforaminal region located at positions 34, 33, 32, 42, 43, 44. Bone level in relation to implant margin was measured and recorded. In order to obtain an in patient comparison of immediately loaded and non-loaded implants, the ones at 33 and 43 were chosen to be immediately loaded by a Dolder-bar retained overdenture. The implants in position 32, 34, 42 and 44 were covered and left to heal. After a healing period of 6 months, second stage surgery was carried out. The clinical criteria to be checked at this point were survival, Periotest values and marginal bone level at the loaded and non-loaded implants. The mean Periotest value was 2.7 for the loaded and -5.6 for the non-loaded implants. The Mann-Whitney U-test showed that the difference was highly significant (P < 0.001). The mean bone level changes at prosthetic delivery were 0.9 mm resorption for the loaded implants and 0.33 mm for non-loaded implants. The difference was highly significant (P < 0.001). No implant failures were observed up to the prosthetic restoration 6 months post insertion. The results of this investigation allowed for direct comparison of implant survival and clinical results between immediately loaded implants and standard implants. Clinical bone changes at the 6 month evaluation demonstrated significantly higher crestal resorption around loaded implants. This fact was confirmed by higher median Periotest values (-3 vs. -6) of immediately loaded implants. According to the outcome of this study, immediate loading of two interforaminal implants with a Dolder-bar resulted in an intimate bone apposition comparable with implants with submerged healing. Nevertheless, the coronal bone level as well as clinical stability (PTV) were significantly lower in the case of the immediately loaded implants. Future studies will be necessary to evaluate marginal bone resorption, Periotest values and clinical success rates of mandibular immediately loaded implants in the long term. PMID- 12755778 TI - Morphology and dimensions of the mandibular jaw bone in the interforaminal region in patients requiring implants in the distal areas. AB - This study aimed to analyse variations in the mandibular interforaminal morphology in an attempt to identify potential risks or contraindications for surgery, especially implant installation, in this particular region. A total of 210 spiral computer tomography (CT) examinations of patients requiring endosseous implant installation in the lower jaw were re-evaluated to explore anatomical variations in bone morphology (shape and contour), and to measure parameters concerning height, width and inclination of the bone in the symphyseal atrea. All measurements were performed on the cross-sectional reformatted images mesial to the mental foramina. A lingual concavity (with a depth of 6 +/- 2.6 mm) was observed in 2.4% of the jaws, with a remaining bone height in that area ranging from 4.2 to 11.9 mm. A clearly lingual tilted/inclined morphology was seen in 28.1% of jaws with a mean angle of 67.6 +/- 6.5 degrees, but a relatively constant width (> 8.8 mm). The remaining jaws (69.5%) showed a slight broadening in the caudal direction. The morphologic parameters were influenced neither by age nor by gender. In conclusion, mandibles with a lingual concavity or a severe slope of the lingual cortex might confer increased risks of lingual perforations during trepanation surgery or graft harvesting as well as fenestrations during implant installation. The detection frequency of such variations within the lower jaw seems to advocate a profound dissection of the lingual site and, in the case of some special treatment strategies, additional cross-sectional radiography. PMID- 12755779 TI - Solcoseryl, a tissue respiration stimulating agent, significantly enhances the effect of capacitively coupled electric field on the promotion of bone formation around dental implants. AB - In the present study we examined the combined effect of application of a capacitively coupled electric field (CCEF) and the tissue respiration stimulating agent, Solcoseryl, on the promotion of bone formation around dental implants histologically and mechanically. After a dental implant was inserted into each femur of Japanese white rabbits, Solcoseryl (2 ml/kg) was administered intravenously in the ear vein and a CCEF was applied for 4 h per day for 14 days. The degree of bone formation on microscopic observation, bone contact ratio, bone surface area ratio, and the level of removal torque of the implant in the Solcoseryl- and CCEF-treated group were significantly higher than the respective value in the control group, which had not been treated with Solcoseryl nor CCEF. Thus, the combination of CCEF stimulation and Solcoseryl effectively promoted the formation of new bone. It is suggested that the clinical use of a combination of CCEF stimulation and Solcoseryl for dental implants promotes osseointegration. PMID- 12755780 TI - Implant-supported fixed cantilever prostheses in partially edentulous arches. A seven-year prospective study. AB - The purpose of the following study was to evaluate the medium- to long-term prognosis of implant-supported cantilever fixed prostheses, and to establish to what degree this is influenced by factors such as length, type of cantilever (mesial or distal), and opposite dentition versus cantilever prostheses. This study was performed on a sample of 38 partially edentulous patients treated between January 1994 and March 2001 with 49 partial cantilever fixed prostheses supported by 100 implants. Marginal bone resorption (MBL) has been studied and used as a reference parameter to define therapeutic success. The MBL measurement was made possible by transposing X-ray images of patients selected on a PC and then using a software program. Statistical analysis was carried out for possible correlation between peri-implant bone resorption and the parameters considered in this study: length and type (mesial or distal) of cantilever and opposite dentition to cantilever prostheses. Seven years after loading cantilever prostheses, the overall cumulative implant survival rate (OCSR) was 97%, and the prostheses success rate is 98%. Mesial cantilever prostheses registered a lower success rate (97.1%) than distal cantilever prostheses (100%). Furthermore, a better prognosis was not observed when the opposite dentition of the prostheses comprised natural teeth, or fixed prostheses on natural teeth, when compared with the cases in which opposite teeth were implant-supported fixed prostheses. The authors concluded that medium-term prognosis of implant-supported cantilever fixed prostheses and traditional implant-supported fixed prostheses was comparable. However, a thorough pre-treatment analysis of risk factors regarding implant-supported prosthesis survival is important. PMID- 12755781 TI - Bone contact, growth, and density around immediately loaded implants in the mandible of mini pigs. AB - The aim of the study was to compare the bone mineral apposition rate (BMAR) of immediately loaded implants with an unloaded control during the early healing phase in the partially edentulous mandible. In seven mini pigs, three premolars and the first molar were removed in the left mandible. Three months later, five implants were installed. Four implants received a fixed provisional restoration and were loaded immediately. The most anterior implant was used as unloaded control. Polychromatic fluorescence labelling was performed to assess the BMAR. After 4 months, the implants were retrieved together with the adjacent bone. Histological specimens were prepared and subjected to a fluorescence microscopic and histomorphometric analysis. Two provisional restorations were found partially lost at the end of the observation period. One implant that had lost the splinting fixation showed soft connective tissue healing. The BMAR did not differ statistically significantly between loaded and unloaded implants and within the single groups during the observation period (BMARloaded days 14-42=1.8+/-0.2 microm/d, BMARloaded days 42-70=1.8+/-0.1 microm/d, BMARloaded days 70-98=1.6+/ 0.1 microm/d, pBMARloaded days 14-42/42-70/70-98 =0.156, BMARunloaded days 14 42=1.7+/-0.1 microm/d, BMARunloaded days 42-70=1.8+/-0.2 microm/d, BMARunloaded days 70-98=1.6+/-0.4 microm/d, pBMARunloaded days 14-42/42-70/70-98=0.368, pBMARloaded/unloaded days 14-42=0.073, pBMARloaded/unloaded days 42-70=0.098, pBMARloaded/unloaded days 70-98=0.262). Four months after implant placement, the bone-to-implant contact was 77.8+/-17.3% for the loaded and 78.0+/-5.8% for the unloaded implants (P=0.753). Immediate loading does not affect the bone mineral apposition rate when compared with unloaded implants. Rigid splinting seems to be the crucial factor for implant success. Uncontrolled masticatory forces can cause failure after partial loss of the provisional restoration. PMID- 12755782 TI - Maxillary sinus floor elevation surgery. A clinical, radiographic and endoscopic evaluation. AB - Although augmentation of the maxillary sinus floor with autogenous bone grafts has become a well established preimplantology procedure, its effect on the function of the maxillary sinus has not been the subject of prospective human studies. In this prospective study the effects of sinus floor augmentation on maxillary sinus performance were evaluated. Seventeen consecutive patients who were to undergo augmentation of the maxillary sinus floor with an iliac crest autogenous bone graft agreed to participate in this study. All patients were subject to (i) extensive anamnestic and clinical investigation on sinusitis, (ii) conventional radiography (Waters' projection) and (iii) unilateral endoscopic inspection of the maxillary sinus. This triad of evaluations was performed preoperatively, immediately preceding the augmentation procedure (the maxillary sinus to be inspected endoscopically was randomly selected), and at 3 (at insertion of the implants) and 9 months (at uncovering of implants) postaugmentation. None of the 17 patients showed clinical or radiological signs of actual sinus pathology preoperatively, though 5 patients had a history of an impeded sinus clearance. By contrast, unilateral endoscopic evaluation revealed pre-existing subclinical mucosal pathology in two out of five patients with a history of sinus clearance impairment and in one out of the other 12 patients. At 3 months' postaugmentation, clinical and radiographical examination showed chronic maxillary sinusitis in one non-compromised patient. Moreover, serial unilateral endoscopic evaluation revealed subclinical maxillary mucosal pathology in four other patients (two of whom had a history of an impeded sinus clearance), confirmed by Waters' projection in three of these four patients. At 9 months' postaugmentation, only subclinical maxillary mucosal pathology was detected endoscopically in two patients (one compromised, one non-compromised patient), confirmed by Waters' projection in this last patient. Five implants were lost during the 9-month observation period. As is obvious from this prospective evaluation, the effects of the augmentation procedure on maxillary sinus performance in patients without signs of maxillary sinusitis are of no clinical significance. PMID- 12755783 TI - Long-term implant prognosis in patients with and without a history of chronic periodontitis: a 10-year prospective cohort study of the ITI Dental Implant System. AB - AIM: The aim of this 10-year study was to compare the failure, success and complication rates between patients having lost their teeth due to periodontitis or other reasons. MATERIAL AND METHODS: Fifty-three patients who received 112 hollow screw implants (HS) of the ITI Dental Implant System were divided into two groups: group A - eight patients with 21 implants having lost their teeth due to chronic periodontitis; group B - forty five patients with 91 implants without a history of periodontitis. One and 10 years after surgical placement, clinical and radiographic parameters were assessed. The incidences of peri-implantitis were noticed over the 10 years of regular supportive periodontal therapy. RESULTS: Success criteria at 10 years were set at: pocket probing depth (PPD) 21 kb) was sequenced from 26 chromosomes from a group of Holstein-Friesian cows, as well as exon 3 of PRNP (>4 kb) from a further 24 chromosomes from six diverse breeds. We identified 51 variant sequences of which 42 were single nucleotide polymorphisms and nine were insertion/deletion (indel) events. The study was extended to exon 3 of the sheep PRNP gene where 23 sequence variants were observed, four of which were indels. The level of nucleotide diversity in the complete bovine PRNP gene was pi = 0.00079, which is similar to that found at the bovine T-cell receptor alpha delta joining region (pi = 0.00077), but somewhat less than that observed for the bovine leptin (pi = 0.00265). Sequence variation within exon 3 of PRNP in both cattle (pi = 0.00102) and sheep (pi = 0.00171) was greater than that for the complete PRNP gene, with sheep showing greater sequence variation in exon 3 than cattle. The level of sequence variation reported here is greater than previously thought for the bovine PRNP gene in cattle. This study highlights the contribution that recombination plays in increasing allelic diversity in this species. PMID- 12755819 TI - Deletion of one of the duplicated Hsp70 genes causes hereditary myopathy of diaphragmatic muscles in Holstein-Friesian cattle. AB - Heat-shock protein 70 (Hsp70) is a major chaperone that folds protein and prevents aggregation. The Hsp70 family contains both constitutive and stress inducible forms. In humans, two of the inducible Hsp70 genes are located within the human major histocompatibility complex (MHC) on 6p21.3, as a duplicated locus, 12 kb apart from each other. We report that loss of one of the duplicated Hsp70 genes, the bovine homologue within the bovine MHC, is responsible for hereditary myopathy of diaphragmatic muscles (HMDM) in Holstein-Friesian cattle. Although the remaining Hsp70 gene is intact, Hsp70 protein levels are dramatically decreased in affected cattle. In normal diaphragmatic muscle, Hsp70 binds several proteins involved in energy metabolism including glycogen phosphorylase (PYGM). Immunohistochemical staining indicated that PYGM accumulated in the HMDM-specific core-like structures in affected cattle. Misfolding of energy-related proteins due to Hsp70 deficiency might lead to protein aggregation and muscle fibre degeneration. PMID- 12755820 TI - A radiation hybrid map of bovine chromosome 24 and comparative mapping with human chromosome 18. AB - We present herein a bovine chromosome 24 (BTA24) radiation hybrid (RH) map using 40 markers scored on a panel of 90 RHs. Of these markers, 29 loci were ordered with odds of at least 1000:1 in a framework map. An average retention frequency of 17.4% was observed, with relatively higher frequencies near the centromere. The length of the comprehensive map was 640 centiray5000 (cR5000) with an average marker interval of approximately 17.3 cR5000. The observed locus order is generally consistent with currently published bovine linkage and physical maps. Nineteen markers were either Type I loci or closely associated with expressed sequences and thus could be used to compare the BTA24 RH map with human mapping information. All genes located on BTA24 were located on human chromosome 18, and previously reported regions of conserved synteny were extended. The comparative data revealed the presence of at least six conserved regions between these chromosomes. PMID- 12755821 TI - Use of a novel outbred by inbred F1 cross to detect genetic markers for growth. AB - A unique outbred by inbred F1 resource population was established. The population structure facilitated the unique opportunity of examining gene by genetic background interaction through crossing two modern broiler sires with dams from two unrelated inbred lines, with no selection for growth rate, to produce about 600 F1 chicks. Pools of DNA were generated from the phenotypic extremes (20% high and low) for 8-week body weight for each of the four combinations of sire and dam line. For one sire family, pools were also separately generated for each sex. The pools were genoyped with 25 informative (segregating) microsatellites. This unique F1 cross between outbred and inbred populations allowed use of the inbred alleles as an 'internal control' for polymerase chain reaction amplification quality in DNA pools. Ten microsatellites showed marked differences (P < 0.05) in allele frequencies between high and low pools, suggesting an association between marker and quantitative trait loci (QTL). These differences were verified using selective genotyping. For many markers, differences in allele frequencies between the high and the low pools, or marker effect, varied between the two dam lines and the two sexes, suggesting an interaction between some genes and the genetic background as represented by different dam lines or sexes. The suggestive marker QTL associations identified in this F1 population demonstrate the efficiency of this population design while different QTL effects in different genetic line crosses and sexes highlight the importance of gene by genetic background interaction in QTL detection. PMID- 12755822 TI - Molecular cloning of the porcine inhibin-betaB gene and reassignment to chromosome 15. AB - Inhibins are gonadal glycoproteins belonging to the transforming growth factor beta superfamily that act to suppress pituitary follicle stimulating hormone and are composed of a common alpha-subunit linked by disulphide bonds to either a betaA- or betaB-subunit. The porcine inhibin-alpha, -betaA (INHBA) and -betaB (INHBB) subunit genes have previously been mapped to chromosomes 15, 18 and 12, respectively. Over 6.7 kb of the INHBB gene was sequenced from a porcine genomic cosmid clone and found to contain two microsatellites, one in intron 1 and the other in the 3'-untranslated region. Both microsatellites mapped to pig chromosome 15 at relative position 48 cm. This sequence was greater than 99% identical to two previously reported partial non-contiguous cDNAs for porcine INHBB. Non-coding regions also had a high degree (79-88%) of identity with the corresponding regions of the human gene. Based on sequence information and mapping of two novel microsatellite markers, we reassigned porcine INHBB to chromosome 15, which is consistent with comparative physical and linkage maps of this chromosome and human chromosome 2. PMID- 12755823 TI - Construction of a new porcine whole-genome framework map using a radiation hybrid panel. AB - We have constructed a radiation hybrid (RH) map of the porcine genome using an RH panel generated by an irradiation dose of 5000-rad (Sus scrofa radiation hybrid map, SSRH map). Normal porcine aortic endothelial cells were irradiated and fused with a thymidine kinase-deficient mouse cell line, L-M (TK-). A total of 110 cell lines were selected and used for further analysis. Among 1091 microsatellite (MS) markers selected for mapping, 842 markers (77%) could be typed on the panel. The framework map comprised 342 MS markers and an additional 247 MS markers were then added to generate the whole-genome map. The average retention frequency for the data set was 30.6%. The total map length was 5596.2 centiRay (cR). Using an estimated physical length of 2718 Mbp, the average ratio between cR and physical distance over the porcine genome was estimated to be 0.49 Mb/cR. PMID- 12755824 TI - Identification of SNPs, mapping and analysis of allele frequencies in two candidate genes for meat production traits: the porcine myosin heavy chain 2B (MYH4) and the skeletal muscle myosin regulatory light chain 2 (HUMMLC2B). AB - Myosin is one of the most important skeletal muscle proteins. It is composed of myosin heavy chains and myosin light chains that exist with different isoforms coded by different genes. We studied the porcine myosin heavy chain 2B (MYH4) and the porcine skeletal muscle myosin regulatory light chain 2 (HUMMLC2B) genes. A single nucleotide polymorphism (SNP), identified for each gene, was used for linkage mapping of MYH4 and HUMMLC2B to porcine chromosome (Sscr) 12 and Sscr 3, respectively. The mapping of these two genes was confirmed by using a porcine rodent radiation hybrid panel, even if for MYH4 the LOD score and the retention fraction were low. Allele frequencies at the two loci were studied in a sample of 307 unrelated pigs belonging to seven different pig breeds. Moreover the distribution of the alleles at these two loci was analysed in groups of pigs with extreme divergent (positive and negative) estimated breeding values (EBV) for four meat production traits that have undergone selection in Italian heavy pigs. PMID- 12755825 TI - Estimation of genetic variability of the founder population in a conservation scheme using microsatellites. AB - In a conservation programme with genealogical records it is possible to estimate the amount of variability of the founder population from a measure of the similarity among the individuals in the current population based on microsatellite markers. Here we compare three available methods and we shown that the one based on the molecular coancestry coefficient should be preferred. PMID- 12755826 TI - Differences in recombination rates on chromosome 23 between German Angus and German Simmental and breed specific linkage mapping. AB - Five paternal half sib families of German Angus (GA) (n = 428) and six of German Simmental (GS) (n = 378) including dams were genotyped with 11 microsatellites (INRA132, RM033, BM1815, BM1258, BOLA-DRB1, BM1818, BM1905, BM1443, CYP21, CSSM5 and DYMS1) derived from chromosome 23. Differences in heterozygosity between the breeds were observed. Significant differences in recombination rates between GA and GS could be demonstrated for the marker intervals INRA132-CSSM5, CYP21-BOLA DRB1 and BOLA-DRB1-BM1818. The length of the map of GA was 90.5 cM in contrast to 117.8 cM for GS. The breed specific linkage maps show differences in length but confirmation of the order of the markers. PMID- 12755827 TI - Linkage and radiation hybrid mapping of the porcine MEF2D gene to chromosome 4q. PMID- 12755828 TI - The bovine type I iodothyronine deiodinase (DIO1) gene maps to chromosome 3. PMID- 12755829 TI - Assignment of the fatty acid synthase (FASN) gene to pig chromosome 12 by physical and linkage mapping. PMID- 12755830 TI - Characterization of a bovine gene encoding an ankyrin repeat and SOCS box protein (ASB15). PMID- 12755831 TI - Eleven previously unreported dinucleotide microsatellite loci on bovine chromosome 19. PMID- 12755832 TI - Chicken syndecan-4 (SDC4) maps to linkage group E32E47W24. PMID- 12755833 TI - Linkage and radiation hybrid mapping of the porcine MYF6 gene to chromosome 5. PMID- 12755834 TI - Review article: Oral, modified-release mesalazine formulations--proprietary versus generic. AB - Products containing mesalazine have been used in the treatment of inflammatory bowel disease for many years. Many of the oral, modified-release products are reaching the point of patent expiration, and it is expected that several new 'generic' versions will be developed. As mesalazine acts topically, the drug needs to be available at the site of inflammation to be effective. For this reason, the currently available products have been developed with individual formulations so that physicians have a choice when matching the different release profiles to the site and extent of disease. As such, the current guidelines state that oral, delayed-release mesalazine formulations are not interchangeable and should be prescribed by their proprietary (brand) name. The standard regulatory assessment process for generic or 'copy' products, using systemic bioequivalence data, does not appear to be sufficient when evaluating topically acting, oral, modified-release products. We therefore recommend that the regulatory bodies should require that new, oral mesalazine products should be assessed by a combination of dissolution, bioequivalence and (a minimum of one) adequately powered, comparative trial to determine therapeutic equivalence. Of most importance here is that the assessment of new modified-release products is sufficiently rigorous to allow patients and physicians to be confident in their use. PMID- 12755835 TI - Systematic review: Antacids, H2-receptor antagonists, prokinetics, bismuth and sucralfate therapy for non-ulcer dyspepsia. AB - BACKGROUND: Evidence for the effectiveness of antacids, histamine-2 receptor antagonists, bismuth salts, sucralfate and prokinetic therapy in non-ulcer dyspepsia is conflicting. AIM: To conduct a systematic review evaluating these therapies in non-ulcer dyspepsia. METHODS: Electronic searches were performed using the Cochrane Controlled Trials Register, Medline, EMBASE, Cinahl and SIGLE until September 2002. Dyspepsia outcomes were dichotomized into cured/improved vs. same/worse. RESULTS: Prokinetics [14 trials, 1053 patients; relative risk reduction (RRR), 48%; 95% confidence interval (95% CI), 27-63%] and histamine-2 receptor antagonists (11 trials, 2164 patients; RRR, 22%; 95% CI, 7-35%) were significantly more effective than placebo. Bismuth salts (RRR, 40%; 95% CI, - 3% to 65%) were superior to placebo, but this was of marginal statistical significance. Antacids and sucralfate were not statistically significantly superior to placebo. A funnel plot suggested that the prokinetic and histamine-2 receptor antagonist results could be due to publication bias. CONCLUSIONS: The meta-analyses suggest that histamine-2 receptor antagonists and prokinetics are superior to placebo. These data are difficult to interpret, however, as funnel plot asymmetry suggests that the magnitude of the effect could be due to publication bias or other heterogeneity-related issues. PMID- 12755836 TI - Meta-analysis: proton pump inhibitor or H2-receptor antagonist for Helicobacter pylori eradication. AB - AIM: To compare H2-receptor antagonists and proton pump inhibitors as adjuvants to triple therapy for Helicobacter pylori eradication. METHODS: H. pylori infected patients with peptic ulcer were randomized to receive either 300 mg nizatidine or 30 mg lansoprazole plus 1 g amoxicillin and 500 mg clarithromycin taken b.d. for 7 days. H. pylori eradication was assessed 4 weeks after therapy. Using meta-analytical techniques, we combined the results of this study with other randomized controlled comparisons of H2-receptor antagonists and proton pump inhibitors as adjuvants to triple therapy. RESULTS: One hundred and one patients were randomized. H. pylori eradication was 94% (47/50) [95% confidence interval (CI), 83-99%] (intention-to-treat) in the H2-receptor antagonist group vs. 86% (44/51) (95% CI, 74-94%) in the proton pump inhibitor group (P = 0.3). There has been a total of 12 similar studies (1415 patients). The overall efficacy was similar in intention-to-treat analysis: 78% (549/701) with H2 receptor antagonists vs. 81% (575/714) with proton pump inhibitors (odds ratio, 0.86; 95% CI, 0.66-1.12). A non-significant trend favouring H2-receptor antagonist (79% vs. 69%; odds ratio, 1.14; 95% CI, 0.76-1.71; P = 0.5) was seen in the comparison of clarithromycin-containing regimens. In contrast, in non clarithromycin-containing trials, there was a slight, but significant, advantage with proton pump inhibitors (85% vs. 78%; odds ratio, 0.64; 95% CI, 0.45-0.92; P = 0.02). CONCLUSION: Overall, proton pump inhibitor and H2-receptor antagonist antisecretory agents appear to be similarly effective as adjuvants for H. pylori triple therapy. It is unlikely that the direct anti-H. pylori effect of proton pump inhibitors is responsible for their ability to enhance anti-H. pylori therapy. PMID- 12755837 TI - Meta-analysis: comparing the efficacy of proton pump inhibitors in short-term use. AB - BACKGROUND: Proton pump inhibitors have a prominent role in the management of acid-related diseases. Controlling expenses on proton pump inhibitors would yield great economic benefits for Dutch health care. AIM: To investigate whether clinical differences in proton pump inhibitors exist. METHODS: We searched Medline, EMBASE and the Cochrane Library. We identified papers in English, German, French or Dutch in which two or more proton pump inhibitors were compared under the same clinical conditions in gastro-oesophageal reflux disease, peptic ulcer disease or Helicobacter pylori eradication. The pooled relative risks were calculated using the Mantel-Haenszel method. RESULTS: Two significant differences were found in the proton pump inhibitors compared. In gastro-oesophageal reflux disease, esomeprazole 40 mg was superior to omeprazole 20 mg (relative risk, 1.18; 95% confidence interval, 1.14-1.23). In peptic ulcer disease, pantoprazole 40 mg was superior to omeprazole 20 mg (relative risk, 1.07; 95% confidence interval, 1.02-1.13). In Helicobacter pylori eradication, no significant differences were found. CONCLUSIONS: Both significant differences found were in favour of the highest dose of proton pump inhibitor on a milligram basis. This indicates that the difference may be dose dependent and not proton pump inhibitor specific. Therefore, when prescribing proton pump inhibitors, arguments other than clinical efficacy, such as those related to pharmaco-economics, may be considered. PMID- 12755838 TI - Meta-analysis: evaluation of adjuvant therapy after curative liver resection for hepatocellular carcinoma. AB - AIM: To evaluate adjuvant modalities after curative resection for hepatocellular carcinoma using a meta-analysis of randomized and non-randomized controlled trials. METHODS: In a first step, a meta-analysis of randomized controlled trials was carried out. Sensitivity analyses after inclusion of non-randomized controlled trials were performed. Four therapeutic modalities were evaluated: pre operative transarterial chemotherapy, post-operative transarterial chemotherapy, systemic chemotherapy and a combination of systemic and transarterial chemotherapy. RESULTS: Only post-operative transarterial chemotherapy improved survival significantly at 2 years [difference, 22.8%; confidence interval (CI), 8.6-36.9%; P = 0.002] and 3 years (difference, 27.6%; CI, 8.2-47.1%; P = 0.005), and decreased the probability of no recurrence at 1 year (difference, 28.8%; CI, 16.7-40.8%; P < 0.001), 2 years (difference, 27.6%; CI, 8.2-47.1%; P = 0.005) and 3 years (difference, 28%; CI, 8.2-47.9%; P = 0.006). In a sensitivity analysis after inclusion of non-randomized controlled trials, post-operative transarterial chemotherapy still improved survival at 1 year (difference, 9.6%; CI, 0.8-18.3%; P = 0.03), 2 years (difference, 13.5%; CI, 0.9-26%, P = 0.04) and 3 years (difference, 18%; CI, 7-28.9%; P < 0.001), and decreased the probability of no recurrence at 1 year (difference, 20.3%; CI, 7.7-33%; P = 0.002), 2 years (difference, 35%; CI, 21.4-46.3%; P < 0.001) and 3 years (difference, 34.5%; CI, 18.7-50.3%; P < 0.001). CONCLUSION: Post-operative transarterial chemotherapy improved survival and decreased the cumulative probability of no recurrence. New randomized controlled trials evaluating this modality are required. PMID- 12755839 TI - Infliximab in refractory pouchitis complicated by fistulae following ileo-anal pouch for ulcerative colitis. AB - AIM: To determine the efficacy of infliximab in the treatment of chronic refractory pouchitis complicated by fistulae following ileal pouch-anal anastomosis for ulcerative colitis. METHODS: This open study included seven patients (four females, three males) with chronic refractory pouchitis complicated by fistulae. Pouchitis was diagnosed by clinical, endoscopic and histological criteria. The sites of the fistulae were as follows: pouch-bladder in one, vaginal in three, perianal in two, and both vaginal and perianal in one. Extra-intestinal manifestations (erythema nodosum, arthralgia) were present in four patients. Crohn's disease was carefully excluded in all patients after re evaluation of the history, re-examination of the original proctocolectomy specimen and examination of the proximal small bowel. All patients had been treated with antibiotics and three with steroids. Patients received infliximab, 5 mg/kg, at 0, 2 and 6 weeks. Azathioprine (2.5 mg/kg) was also started for all patients as bridge therapy. Clinical response was classified as complete, partial or no response. Fistulae closure was classified as complete (cessation of fistulae drainage and total closure of all fistulae), partial (a reduction in the number, size, drainage or discomfort associated with fistulae) or no closure. The pouchitis disease activity index and quality of life were also used as outcome measures. RESULTS: Clinically, all patients improved. At the 10-week follow-up, six of the seven patients had a complete clinical response, and five had complete fistulae closure. At the 10-week follow-up, the median pouchitis disease activity index decreased from 12 (baseline) (range, 10-15) to 5 (range, 3-8); the median quality of life decreased from 37 points (range, 33-40) to 14 (range, 9-18). Erythema nodosum and arthralgia showed complete remission soon after the first infusion of infliximab. CONCLUSIONS: These preliminary results indicate that infliximab may be recommended for the treatment of refractory pouchitis complicated by fistulae following ileal pouch-anal anastomosis for ulcerative colitis. PMID- 12755840 TI - Rescue therapy with tacrolimus is effective in patients with severe and refractory inflammatory bowel disease. AB - BACKGROUND: Oral tacrolimus, approved for the prophylaxis of organ rejection in liver or kidney transplants, has been reported to be effective in anecdotal cases of refractory inflammatory bowel disease. AIM: To evaluate the usefulness of low dose oral tacrolimus in refractory inflammatory bowel disease. METHODS: Thirty one adult Caucasian patients with steroid-dependent (n = 15) or steroid refractory (n = 16) inflammatory bowel disease (Crohn's disease, n = 6; ulcerative colitis, n = 23; pouchitis, n = 2) were enrolled. Tacrolimus (0.1 mg/kg body weight per day) was administered orally in 30 patients and initially intravenously in one patient (0.01 mg/kg body weight per day), aiming for serum trough levels of 4-6 ng/mL. The median treatment duration was 12 months (range, 1 137 months). RESULTS: Twenty-eight patients (90.3%) experienced a clinical and laboratory response and 20 (64.5%) went into remission. One ulcerative colitis patient and two Crohn's disease patients did not improve. Three ulcerative colitis patients (9.7%) were colectomized at 1, 12 and 24 months after tacrolimus initiation. In 19 of 23 patients (82.6%) taking steroids, steroids were reduced or discontinued. Side-effects included a temporary rise of creatinine (n = 3, 9.7%), tremor or paraesthesias (n = 3, 9.7%), hyperkalaemia (n = 1, 3.2%), hypertension (n = 1, 3.2%) and an opportunistic infection (n = 1, 3.2%). CONCLUSION: Oral tacrolimus is safe and effective in refractory inflammatory bowel disease. PMID- 12755841 TI - The selection of triple therapy for Helicobacter pylori eradication in chronic renal insufficiency. AB - AIM: To establish a triple therapy regimen for Helicobacter pylori eradication in patients with chronic renal insufficiency. METHODS: Eighty-eight patients with chronic renal insufficiency and H. pylori infection were evenly randomized into two groups receiving 1-week lansoprazole, 30 mg, clarithromycin, 500 mg, and either amoxicillin, 750 mg, or metronidazole, 500 mg, twice daily. The adverse events and compliance with triple therapy were reviewed at the week 1 visit. Patients provided stool samples at week 6 to assess the success of H. pylori eradication by H. pylori-specific stool antigen. The serum creatinine levels were monitored at enrollment, at weeks 1, 2 and 6 and on any unscheduled visit after triple therapy. RESULTS: The success of H. pylori eradication was higher in the lansoprazole-clarithromycin-metronidazole group than in the lansoprazole clarithromycin-amoxicillin group (intention-to-treat analysis: 84% vs. 66%, P < 0.05: per protocol analysis: 93% vs. 76%, P < 0.05). Complete drug compliance was also better in the lansoprazole-clarithromycin-metronidazole group than in the lansoprazole-clarithromycin-amoxicillin group (77% vs. 52%, P < 0.05). Patients in the lansoprazole-clarithromycin-metronidazole group had a lower risk of acute renal failure than those in the lansoprazole-clarithromycin-amoxicillin group (2% vs. 18%, P < 0.05; relative risk, 0.128, 95% confidence interval, 0.016-0.979). CONCLUSIONS: Triple therapy with metronidazole and clarithromycin, but not amoxicillin, can be used for H. pylori eradication in patients with chronic renal insufficiency, because it is more effective, well tolerated and less likely to cause deterioration of renal function. PMID- 12755843 TI - Clinical usefulness of KRAS mutational analysis in the diagnosis of pancreatic adenocarcinoma by means of endosonography-guided fine-needle aspiration biopsy. AB - AIM: To establish the usefulness of KRAS mutational analysis in the diagnosis of pancreatic adenocarcinoma by comparing this technique with conventional cytology in aspirates obtained by endosonography-guided fine-needle aspiration. METHODS: All consecutive patients with pancreatic focal lesions undergoing endosonography guided fine-needle aspiration were included. Samples were obtained with the concurrence of an attendant cytopathologist. Detection of codon-12 KRAS mutations was performed by the restriction fragment length polymorphism-polymerase chain reaction method. The effectiveness of conventional cytology, KRAS mutational analysis and their combination was established with respect to the definitive diagnosis. A cost-effectiveness analysis was also performed. RESULTS: Thirty three patients had pancreatic adenocarcinoma and 24 patients had other lesions. A total of 136 samples was obtained. In patients in whom specimens were adequate (93% for cytology; 100% for mutational analysis), the specificity of both techniques was 100%, whereas the sensitivity favoured cytology (97% vs. 73%). When inadequate samples were considered as misdiagnosed, a combination of both techniques reached the highest overall accuracy (cytology, 91%; mutational analysis, 84%; combination of both, 98%). CONCLUSIONS: Cytology from aspirates obtained by endosonography-guided fine-needle aspiration is the most precise single technique for the diagnosis of pancreatic adenocarcinoma. However, when adequate specimens are not available to reach a cytological diagnosis, the addition of KRAS mutational analysis represents the best strategy. PMID- 12755842 TI - An inter- and intra-laboratory comparison of breath 13CO2analysis. AB - BACKGROUND: 13C breath test analysis requires accurate 13CO2measurements. AIM: To perform a multicentre study to evaluate the repeatability and reproducibility of breath 13CO2analysis. METHODS: Two series of 25 paired randomly coded tubes (each consisting of 23 13CO2-enriched breath samples and two samples of standard reference pure CO2with certified delta 13C(PDB)) were sent to participating centres for 13CO2measurement. Each series of tubes was analysed 10 days apart. The repeatability and reproducibility of 13C measurements was assessed by Mandel's k and h statistics. RESULTS: Twenty-two centres participated in the study: 18 showed good inter- and intra-laboratory variability, whilst four showed abnormally high inter- or intra-laboratory variability. Breath test results were also significantly affected by the accuracy of the 13C analytical procedures. CONCLUSIONS: A low accuracy of 13C measurements may significantly affect the results of breath tests, leading to inappropriate clinical decisions. Standardization of 13C analysis is required to guarantee optimal 13C measurements and accurate 13C breath test results. PMID- 12755845 TI - Current practice in surveillance strategy for patients with Barrett's oesophagus in the UK. AB - BACKGROUND: Many guidelines exist regarding the surveillance of patients with Barrett's oesophagus. There are limited data, however, with regard to whether practitioners follow these guidelines. METHODS: We assessed current surveillance practice amongst members of the British Society of Gastroenterology using a simple 11-question anonymous survey, mailed to 300 randomly selected members from the British Society of Gastroenterology Handbook. RESULTS: Two hundred and three of the 300 (68%) responded, 76% considering that surveillance was worthwhile. In those who considered surveillance to be worthwhile, 83% used sub-selection based on age, the length of Barrett's oesophagus or the presence of ulcer or stricture. Patients with Barrett's oesophagus of < 3 cm (short-segment) were considered to be inappropriate for surveillance by 62%. Forty-one per cent reported following the 'advised' recommendations of four-quadrant biopsies every 2 cm, whereas 44% followed a 'random and suspicious areas only' protocol. Marked variation was reported in the re-endoscope interval for both low- and high-grade dysplasia. Only 55% reported that two experienced pathologists reviewed all biopsies showing high-grade dysplasia. CONCLUSIONS: Despite the existence of multiple guidelines for Barrett's surveillance, clinical practice varies widely in the UK. This may be due to a lack of knowledge or because gastroenterologists remain unconvinced by the quality of the current evidence of its value. These results have implications for studies attempting to collate data from multiple centres. PMID- 12755844 TI - The burden of illness of gastro-oesophageal reflux disease: impact on work productivity. AB - BACKGROUND: The impact of gastro-oesophageal reflux disease on work productivity has become increasingly important, as the symptoms of gastro-oesophageal reflux disease affect individuals in their productive years of life. AIMS: To assess the impact of gastro-oesophageal reflux disease on reduced work productivity and to identify the predictors of reduced productivity. METHODS: A sample of employed individuals reporting chronic heartburn was selected from US household mail survey respondents. Heartburn severity and frequency were recorded using a diary, and work productivity was assessed using the Work Productivity and Activity Impairment Questionnaire for Patients with Symptoms of Gastro-oesophageal Reflux Disease. Predictors of reduced productivity were evaluated. RESULTS: Over 30% of heartburn sufferers reported reduced productivity. Individuals with symptoms of gastro-oesophageal reflux disease (n = 1003) reported 6.0% reduced productivity attributable to symptoms. Over 48% of respondents with severe symptoms reported reduced productivity, compared with 40% and 12% of respondents with moderate and mild symptoms, respectively. Using logistic regression, severity, a younger age and nocturnal symptoms were associated with increased odds of reduced productivity. In those reporting nocturnal heartburn, medication use and sleep interference increased the odds of reduced productivity. CONCLUSIONS: Reduced work productivity is seen in a large proportion of subjects on prescription medication for gastro-oesophageal reflux disease. Symptom severity and nocturnal heartburn are significantly associated with reduced work productivity, particularly when nocturnal heartburn interferes with sleep. PMID- 12755846 TI - Refeeding syndrome: effective and safe treatment with Phosphates Polyfusor. AB - BACKGROUND: Severe hypophosphataemia associated with refeeding syndrome requires treatment with intravenous phosphate to prevent potentially life-threatening complications. However, evidence for replacement regimens is limited and current regimens are complex and replace phosphate inadequately. AIM: To assess the effectiveness and safety of 50 mmol intravenous phosphate infusion, given as a 'Phosphates Polyfusor', for the treatment of severe hypophosphataemia in refeeding syndrome. METHODS: Patients with refeeding syndrome and normal renal function received a Phosphates Polyfusor infusion for the treatment of severe hypophosphataemia (< 0.50 mmol/L). The outcome measures were serial serum phosphate, creatinine and calcium concentrations for 4 days following phosphate infusion and adverse events. RESULTS: Over 2 years, 30 patients were treated. Following treatment, 37% of cases had a normal serum phosphate concentration and 73% had a serum phosphate concentration of > 0.5 mmol/L within 24 h. Ten patients required more than one Phosphates Polyfusor infusion. Within 72 h, 93% of cases had achieved a serum phosphate concentration of > or = 0.50 mmol/L. No patient developed renal failure. Three episodes of transient mild hyperphosphataemia were recorded. Four patients developed mild hypocalcaemia. CONCLUSIONS: This is the largest published series of the use of intravenous phosphate for the treatment of severe hypophosphataemia (< 0.50 mmol/L), and is the most effective regimen described. All patients had refeeding syndrome and were managed on general wards. PMID- 12755849 TI - The European Schizophrenia Outpatient Health Outcomes Study: baseline findings across country and treatment. AB - OBJECTIVE: To describe the baseline findings and study population of the Schizophrenia Outpatient Health Outcomes (SOHO) Study. METHOD: The SOHO study is an ongoing, large, prospective, long-term observational study of schizophrenia treatment in 10 European countries. The study population consists of out-patients who initiate therapy or change to a new antipsychotic. RESULTS: A total of 1096 investigators enrolled 10 972 patients. Approximately 60% of patients were men and the mean age was 40 years. Patients treated with clozapine and more than one antipsychotic are more severely ill, patients receiving depot medications have a history of non-compliance, and patients receiving their first antipsychotic for schizophrenia are most likely to receive an atypical agent. CONCLUSION: The SOHO study population appears to represent European out-patients with schizophrenia in whom a treatment decision is required. Baseline findings reflect European clinical practice with respect to patients treated with individual antipsychotics. PMID- 12755851 TI - A Rasch model analysis to test the cross-cultural validity of the EuroQoL-5D in the Schizophrenia Outpatient Health Outcomes Study. AB - OBJECTIVE: To investigate the pan-European, cross-cultural validity of the EuroQol-5D (EQ-5D) for assessing quality of life in the Schizophrenia Outpatient Health Outcomes (SOHO) Study. METHOD: The EQ-5D items investigated were mobility, self-care, usual activities, pain/discomfort and anxiety/depression. A Rasch rating scale model (a form of differential item functioning) was used to identify invariance of item calibrations for the 10 European countries participating in the SOHO study. RESULTS: There was general congruence in the EQ-5D item calibration pattern. The rank of average EQ-5D item calibrations was similar for all countries except Denmark. Denmark showed slight misfits for mobility and pain/discomfort. CONCLUSION: The EQ-5D is an appropriate measure of health related quality of life across European countries and translations. PMID- 12755850 TI - The Clinical Global Impression-Schizophrenia scale: a simple instrument to measure the diversity of symptoms present in schizophrenia. AB - OBJECTIVE: To describe the development and validation of the Clinical Global Impression-Schizophrenia (CGI-SCH) scale, designed to assess positive, negative, depressive and cognitive symptoms in schizophrenia. METHOD: The CGI-SCH scale was adapted from the CGI scale. Concurrent validity and sensitivity to change were assessed by comparison with the Positive and Negative Symptom Severity (PANSS) and Global Assessment of Functioning (GAF) scales. To evaluate inter-rater reliability, all patients were assessed by two clinicians. RESULTS: Symptoms were assessed in 114 patients. Correlation coefficients between the CGI-SCH and the GAF and PANSS scores were high (most above 0.75), and were highest for positive and negative symptoms. Reliability was substantial (intraclass correlation coefficient, ICC > 0.70) in all but one dimension (depressive dimension, ICC = 0.64). CONCLUSION: The CGI-SCH scale is a valid, reliable instrument to evaluate severity and treatment response in schizophrenia. Given its simplicity, brevity and clinical face validity, the scale is appropriate for use in observational studies and routine clinical practice. PMID- 12755852 TI - Financing health care in Europe: context for the Schizophrenia Outpatient Health Outcomes Study. AB - OBJECTIVE: Exploration of the implications for mental health care of the health care funding environment in Europe, as context for the Schizophrenia Outpatient Health Outcomes (SOHO) Study. METHOD: Data on health care financing for individual countries were sourced by review of the literature and personal communication with European country representatives. RESULTS: The main and complementary sources of health care finance are presented for the 10 European countries participating in the SOHO study. CONCLUSION: A mixture of tax and social insurance funding mechanisms dominate general health care funding in Europe. These mechanisms in principle promote equity in access to all health care interventions, including those for mental health. However, current resource allocation to mental health care may not reflect fully the impact of mental health disorders. Further work is now under way to examine the contribution of non-health sectors (such as social welfare) to the funding and provision of services. PMID- 12755853 TI - Unit costs in international economic evaluations: resource costing of the Schizophrenia Outpatient Health Outcomes Study. AB - OBJECTIVE: We present unit costs corresponding to resource information collected in the Schizophrenia Outpatient Health Outcomes (SOHO) Study. METHOD: The SOHO study is a 3-year, prospective, observational study of health outcomes associated with antipsychotic treatment in out-patients treated for schizophrenia. The study is being conducted across 10 European countries (Denmark, France, Germany, Greece, Ireland, Italy, the Netherlands, Portugal, Spain and the UK) and includes over 10,800 patients and over 1000 investigators. To identify the best available unit costs of hospital admissions, day care and psychiatrist out-patient visits, a tariff-based approach was used. RESULTS: Unit costs were obtained for nine of the 10 countries and were adjusted to 2000 price levels by consumer price indices and converted to US dollars using purchasing power parity rates (and on to Euro). CONCLUSION: The paper illustrates the need to balance the search for sound unit costs with pragmatic solutions in the costing of international economic evaluations. PMID- 12755856 TI - A Chinese definition of nursing. PMID- 12755857 TI - Snap-shots of live theatre: the use of photography to research governance in operating room nursing. AB - The use of photography is an underreported method of research in the nursing literature. This paper explores its use in an ethnographic research project, the fieldwork of which was undertaken by the first author. The aim was to examine the governance of operating room nursing in the clinical setting and the theoretical orientation was the work of Michel Foucault. The focus of this paper is on how photography was used as a means of data generation. To establish some context we begin by drawing on writers from sociology and anthropology to provide an overview of the status of vision and visual research methods in contemporary social research. We then move to a brief discussion of the uses of photography in social research and the limitations imposed by ethical considerations of its use in clinical nursing settings. As well, the process and approach involved in this research project, and issues of analysis are discussed. Three 'snap-shots' of operating room nursing, taken by participants, are presented. Each is analysed in terms of its contributions to the research process as well as its substantive contribution to the theoretical framework and the research aims. PMID- 12755858 TI - Implementing a postcolonial feminist perspective in nursing research related to non-Western populations. AB - In this article, I argue that implementing a postcolonial feminist perspective in nursing research transcends the limitations of modern cultural theories in exploring the health problems of non-Western populations. Providing nursing care in pluralist countries like Canada remains a challenge for nurses. First, nurses must reflect on their ethnic background and stereotypes that may impinge on the understanding of cultural differences. Second, dominant health ideologies that underpin nurses' everyday practice and the structural barriers that may constrain the utilization of public healthcare services by non-Western populations must be further examined. Postcolonial feminism is aimed at addressing health inequities stemming from social discriminative practices. I will draw on extant literature and data of an ongoing ethnography exploring the Haitian caregivers' ways of caring for ageing relatives at home to unveil how the larger social and cultural world has an impact on caregivers' everyday lives. Marginalized locations represent privileged sites from which health problems, intersecting with power, race, gender, and social classes, can be addressed. Postcolonial feminism provides the analytic lens to look at the impact of these factors in shaping health experiences. It also suggests redirecting nursing cultural research and practice to achieve social justice in the healthcare system. PMID- 12755859 TI - Creating what sort of professional? Master's level nurse education as a professionalising strategy. AB - This paper reports on a detailed analysis of selected findings from a larger study of master's level nurse education. It locates some features of such education within the contemporary situation of nursing as a profession and interprets the role of master's level nurse education as a professionalising strategy. In-depth interviews were undertaken with a purposive sample of 18 nurse lecturers drawn from eight universities in the United Kingdom. The interview agenda explored participants' perspectives of the characteristics of master's level performance in practice. Interview transcripts were interpreted by drawing upon hermeneutic methodology. The following themes emerged. (a) The credibility of the master's level nurse was of central importance. In terms of the literature of professionalisation, this may be interpreted as a factor in enhancing the legitimacy of nursing as an occupation. (b) The clinical capability attributed to the nurse is interpreted as leading to an increase in the authority commanded by the expert professional. Thus, the individual capability of the master's level nurse enhances the attribution of autonomous skill to the occupation as a whole. (c) The master's level nurse is seen to exercise influence and leadership and this strengthens the power and status of nursing. Nursing does not have the appearance of a 'traditional' profession, neither has it a clear stance as a 'new profession'. Rather it appears to be especially responsive to the tide of public opinion manifest through government edicts. While nursing is employing rhetoric that espouses both positions, the direction of master's level education is anomalous. PMID- 12755860 TI - Hermeneutic research in nursing: developing a Gadamerian-based research method. AB - This paper takes the stance that although there are many different approaches to phenomenological and hermeneutic research, some of these have become blurred due to multiple interpretations of translated materials. Working from original texts by the German philosophers, this paper reconsiders the relevance of phenomenology and hermeneutics to nursing research. We trace the development of Gadamer's philosophy in order to propose a research method based in this tradition. Five steps have been identified as a guide for nurse researchers. These are deciding upon a question, identification of preunderstandings, gaining understanding through dialogue with participants, gaining understanding through dialogue with text and establishing trustworthiness. PMID- 12755861 TI - Narrative inquiry in a nursing practicum. AB - One approach to creating research-based nursing education is to think and write narratively about the daily life of a BScN program student and her teacher in diverse settings and over time. Gail, as a nurse-teacher, and Faith, as a nursing student and now Public Health Nurse, reconstruct their teaching-learning experiences in an integrated practicum in maternal-child health services as a narrative inquiry. After presenting this reconstruction of experience at a conference on maternal scholarship, further inquiry into their experiences shows how narrative inquiry matters to construction of nursing praxis and to life-long learning as a nurse. Teaching-learning relationships are seen as a template for a student's connections to people experiencing nursing care and to other clinicians. Construction of stories to live by that take into account becoming a nurse, constructing knowledge and enacting caring-healing nursing practices is illuminated through narrative inquiry. PMID- 12755862 TI - The paradox of the Aged Care Act 1997: the marginalisation of nursing discourse. AB - This paper examines the marginalisation of nursing discourse, which followed the enactment of the Aged Care Act 1997. This neo-reform period in aged care, dominated by theories of economic rationalism, enshrined legislation based upon market principles and by implication, the provision of care at the cheapest possible price. This paper exposes some of the gaps in the neo-reform period and challenges the assertion that the amalgamation of nursing homes and hostels in such an environment can provide better quality of care and life for residents. It argues that this amalgamation entails a transformation towards a social model of care and fails to address the professional healthcare needs of the acutely sick and complex extreme old person and makes evident new gaps in the provision of aged care services. The paper proceeds to present strategies where the future for nursing practice in aged care necessarily involves a judicious balancing of individual cases alongside economic prescriptions of care and ever-changing public policy initiatives. It concludes that this can be achieved through a more interactive public, professional and advocacy discourse. The methodology involves extensive analysis of public documents including media, academic journals, government reports and interviews with recognised leaders in the field of aged care. The study utilises a critical interpretative framework consistent with the logic of Michel Foucault. PMID- 12755863 TI - Response to Dave Holmes and Cary Federman: Killing for the state: the darkest side of American nursing. PMID- 12755865 TI - How much effort is required to isolate nuclear microsatellites from plants? AB - The attributes of codominance, reproducibility and high resolution have all contributed towards the current popularity of nuclear microsatellites as genetic markers in molecular ecological studies. One of their major drawbacks, however, is the development phase required to obtain working primers for a given study species. To facilitate project planning, we have reviewed the literature to quantify the workload involved in isolating nuclear microsatellites from plants. We highlight the attrition of loci at each stage in the process, and the average effort required to obtain 10 working microsatellite primer pairs. PMID- 12755866 TI - Population substructures in the soil invertebrate Orchesella cincta, as revealed by microsatellite and TE-AFLP markers. AB - Microsatellite and three enzyme-amplified fragment length polymorphism (TE-AFLP) DNA markers were used to describe the population genetic structure in the soil dwelling collembolan Orchesella cincta (L.). Two forests were sampled according to a three-level nested hierarchical design, with fixed distances among samples within a parcel and among parcels within a forest. The largest component of variation was found at the smallest scale, within parcels (77-97%), while the smallest component of variation was found between forests. The two different methods to study population structure indicated a similar allocation of variance. Population genetic substructuring was revealed between samples on a scale of 50 m; the degree of substructuring however, varied between parcels and forests. One forest showed a high degree of structure as revealed by microsatellites, while another showed a low degree of structure. A significant deviation from random mating (average FIS = 0.23) over the two forests was detected. Two of 18 samples showed a difference in population genetic structure between males and females. We discuss the fact that the population genetic structure of O. cincta is significantly affected by long-range dispersal, even though it is a small and wingless insect. This interpretation is supported by observations on tree climbing behaviour in this species that may facilitate air dispersal. As a consequence, the assumption that migration a priori may be neglected in demographic analysis of O. cincta is incorrect. PMID- 12755867 TI - Patch establishment and development of a clonal plant, Polygonum cuspidatum, on Mount Fuji. AB - Microsatellite analysis was used to investigate the patch establishment and development of Polygonum cuspidatum Sieb. et Zucc, a clonal herbaceous plant that dominates the primary succession on the southeast slope of Mount Fuji. Genotypes of P. cuspidatum in 155 patches at the study site differed from each other. This indicates that P. cuspidatum patches are initially established by seed dispersed on the bare scoria field, and not by clonal rhizome extension. Genetic differentiation was estimated using the FST values between subpopulations at the study site. There was almost no genetic differentiation between subpopulations, indicating the presence of massive gene flow. The pollen fathers of seeds and maternal genets of current-year seedlings were inferred from the microsatellite allele composition by a simple exclusion method. The wide, random distribution of pollen fathers suggests that pollen dispersal occurs over a broad area. Maternal analysis showed a tendency for seed dispersal to be biased to the area nearby and down slope from the mother plants. Patch establishment under massive gene flow may result from such pollen and seed dispersal. To understand the process of patch development, aerial photographs taken from 1962 to 1999 were compared, and then genets in each of 36 patches were identified from the microsatellite genotypes of P. cuspidatum shoots. The comparison of aerial photographs showed that most of the patches enlarged each year and that some neighbouring patches combined during growth. Genet analysis demonstrated a high correlation between patch area and the area of the largest genet within it, and that new genets were recruited at the patch periphery. These findings indicate that both vegetative and sexual reproduction, i.e. rhizome extension and the establishment of new seedlings, contribute to the development of P. cuspidatum patches. PMID- 12755868 TI - An empirical exploration of data quality in DNA-based population inventories. AB - I present data from 21 population inventory studies - 20 of them on bears - that relied on the noninvasive collection of hair, and review the methods that were used to prevent genetic errors in these studies. These methods were designed to simultaneously minimize errors (which can bias estimates of abundance) and per sample analysis effort (which can reduce the precision of estimates by limiting sample size). A variety of approaches were used to probe the reliability of the empirical data, producing a mean, per-study estimate of no more than one undetected error in either direction (too few or too many individuals identified in the laboratory). For the type of samples considered here (plucked hair samples), the gain or loss of individuals in the laboratory can be reduced to a level that is inconsequential relative to the more universal sources of bias and imprecision that can affect mark-recapture studies, assuming that marker systems are selected according to stated guidelines, marginal samples are excluded at an early stage, similar pairs of genotypes are scrutinized, and laboratory work is performed with skill and care. PMID- 12755870 TI - Are the native giant tortoises from the Seychelles really extinct? A genetic perspective based on mtDNA and microsatellite data. AB - The extinction of the giant tortoises of the Seychelles Archipelago has long been suspected but is not beyond doubt. A recent morphological study of the giant tortoises of the western Indian Ocean concluded that specimens of two native Seychelles species survive in captivity today alongside giant tortoises of Aldabra, which are numerous in zoos as well as in the wild. This claim has been controversial because some of the morphological characters used to identify these species, several measures of carapace morphology, are reputed to be quite sensitive to captive conditions. Nonetheless, the potential survival of giant tortoise species previously thought extinct presents an exciting scenario for conservation. We used mitochondrial DNA sequences and nuclear microsatellites to examine the validity of the rediscovered species of Seychelles giant tortoises. Our results indicate that the morphotypes suspected to represent Seychelles species do not show levels of variation and genetic structuring consistent with long periods of reproductive isolation. We found no variation in the mitochondrial control region among 55 individuals examined and no genetic structuring in eight microsatellite loci, pointing to the survival of just a single lineage of Indian Ocean tortoises. PMID- 12755869 TI - Estimating population sizes for elusive animals: the forest elephants of Kakum National Park, Ghana. AB - African forest elephants are difficult to observe in the dense vegetation, and previous studies have relied upon indirect methods to estimate population sizes. Using multilocus genotyping of noninvasively collected samples, we performed a genetic survey of the forest elephant population at Kakum National Park, Ghana. We estimated population size, sex ratio and genetic variability from our data, then combined this information with field observations to divide the population into age groups. Our population size estimate was very close to that obtained using dung counts, the most commonly used indirect method of estimating the population sizes of forest elephant populations. As their habitat is fragmented by expanding human populations, management will be increasingly important to the persistence of forest elephant populations. The data that can be obtained from noninvasively collected samples will help managers plan for the conservation of this keystone species. PMID- 12755871 TI - Was there a second adaptive radiation of giant tortoises in the Indian Ocean? Using mitochondrial DNA to investigate speciation and biogeography of Aldabrachelys (Reptilia, Testudinidae). AB - A radiation of five species of giant tortoises (Cylindraspis) existed in the southwest Indian Ocean, on the Mascarene islands, and another (of Aldabrachelys) has been postulated on small islands north of Madagascar, from where at least eight nominal species have been named and up to five have been recently recognized. Of 37 specimens of Madagascan and small-island Aldabrachelys investigated by us, 23 yielded significant portions of a 428-base-pair (bp) fragment of mitochondrial (cytochrome b and tRNA-Glu), including type material of seven nominal species (A. arnoldi, A. dussumieri, A. hololissa, A. daudinii, A. sumierei, A. ponderosa and A. gouffei). These and nearly all the remaining specimens, including 15 additional captive individuals sequenced previously, show little variation. Thirty-three exhibit no differences and the remainder diverge by only 1-4 bp (0.23-0.93%). This contrasts with more widely accepted tortoise species which show much greater inter- and intraspecific differences. The non Madagascan material examined may therefore only represent a single species and all specimens may come from Aldabra where the common haplotype is known to occur. The present study provides no evidence against the Madagascan origin for Aldabra tortoises suggested by a previous molecular phylogenetic analysis, the direction of marine currents and phylogeography of other reptiles in the area. Ancient mitochondrial DNA from the extinct subfossil A. grandidieri of Madagascar differs at 25 sites (5.8%) from all other Aldabrachelys samples examined here. PMID- 12755872 TI - High multiple paternity and low last-male sperm precedence in a hermaphroditic planarian flatworm: consequences for reciprocity patterns. AB - It is difficult to predict a priori how mating success translates into fertilization success in simultaneous hermaphrodites with internal fertilization. Whereas insemination decisions will be determined by male interests, fertilization will depend on female interests, possibly leading to discrepancies between insemination and fertilization patterns. The planarian flatworm Schmidtea polychroa, a simultaneous hermaphrodite in which mating partners trade sperm was studied. Sperm can be stored for months yet individuals mate frequently. Using microsatellites, maternity and paternity data were obtained from 748 offspring produced in six groups of 10 individuals during four weeks. Adults produced young from four mates on average. Reciprocal fertilization between two mates was found in only 41 out of 110 registered mate combinations, which is clearly less than what is predicted from insemination patterns. Multiple paternity was high: > 80% of all cocoons had two to five fathers for only three to five offspring per cocoon. Because animals were collected from a natural population, 28% of all hatchlings were sired by unknown sperm donors in the field, despite a 10-day period of acclimatization and within-group mating. This percentage decreased only moderately throughout the experiment, showing that sperm can be stored and used for at least a month, despite frequent mating and sperm digestion. The immediate paternity a sperm donor could expect to obtain was only about 25%. Male reproductive success increased linearly with the number of female partners, providing support for Bateman's principle in hermaphrodites. Our results suggest that hermaphrodites do not trade fertilizations when trading sperm during insemination, lending support to the view that such conditional sperm exchange is driven by exchange of resources. PMID- 12755873 TI - Phylogeography of the Northern short-tailed shrew, Blarina brevicauda (Insectivora: Soricidae): past fragmentation and postglacial recolonization. AB - Blarina brevicauda is distributed across the northeastern region of North America, in areas previously covered by Pleistocene glaciers. Previous molecular systematic study of the species in the genus Blarina suggested the presence of two distinct eastern and western phylogroups within B. brevicauda, in agreement with traditionally recognized semi-species. To expand the previous work, a collection of 76 individuals from 14 localities collected throughout the range of B. brevicauda was used to assess the mitochondrial (mt) cytochrome b genealogy for this species. Minimum evolution, maximum parsimony, analysis of molecular variance and nested clade analysis each supported the same conclusions of two well-differentiated and monophyletic east-west groups, separated by the Mississippi River. Denser sampling in areas immediately East of the Mississippi basin revealed further subdivision within the eastern phylogroup into an East Central and an Appalachian clade. The western phylogroup differed from the eastern phylogroup by 2.5% mean absolute DNA sequence difference. About 65% of the genetic variance among samples was explained by the east-west subdivision alone. High haplotype diversities, low nucleotide diversities and unimodal mismatch distributions within subclades suggest recent expansion or diversification within each group. No phylogeographic structure was found within the western phylogroup, but genetic structure because of restricted gene flow and isolation by distance was inferred for the eastern group. The present distribution of B. brevicauda is best explained by past fragmentation and range expansion events during and following the Pleistocene glacial cycles. PMID- 12755874 TI - Chloroplast DNA phylogeography reveals colonization history of a Neotropical tree, Cedrela odorata L., in Mesoamerica. AB - Spanish Cedar (Cedrela odorata L.) is a globally important timber species which has been severely exploited in Mesoamerica for over 200 years. Using polymerase chain reaction-restriction fragment length polymorphisms, its chloroplast (cp) DNA phylogeography was studied in Mesoamerica with samples from 29 populations in six countries. Five haplotypes were characterized, phylogenetically grouped into three lineages (Northern, Central and Southern). Spatial analysis of ordered genetic distance confirmed deviation from a pattern of isolation by distance. The geographically proximate Northern and Central cpDNA lineages were genetically the most differentiated, with the Southern lineage appearing between them on a minimum spanning tree. However, populations possessing Southern lineage haplotypes occupy distinct moist habitats, in contrast to populations possessing Northern and Central lineage haplotypes which occupy drier and more seasonal habitats. Given the known colonization of the proto-Mesoamerican peninsula by South American flora and fauna prior to the formation of the Isthmus of Panama, it seems most likely that the observed population structure in C. odorata results from repeated colonization of Mesoamerica from South American source populations. Such a model would imply an ancient, pre-Isthmian colonization of a dry-adapted type (possessing the Northern lineage or a prototype thereof), with a secondary colonization via the land bridge. Following this, a more recent (possibly post Pleistocene) expansion of moist-adapted types possessing the Southern lineage from the south fits the known vegetation history of the region. PMID- 12755875 TI - Differential patterns of hybridization and introgression between the swallowtails Papilio machaon and P. hospiton from Sardinia and Corsica islands (Lepidoptera, Papilionidae). AB - Proportions of hybridization and introgression between the swallowtails Papilio hospiton, endemic to Sardinia and Corsica, and the holarctic Papilio machaon, were characterized using nine fully diagnostic and two differentiated allozyme loci and a mitochondrial DNA marker. Very low frequencies of F1 hybrids were detected in both Sardinia (0-4%, average 1.4%) and Corsica (0-3%, average 0.5%), as well as of first generation backcrosses (B1). No F2 were observed, in agreement with the hybrid breakdown detected in laboratory crosses. In spite of this minimal current gene exchange, specimens carrying introgressed alleles were found in high proportions in P. machaon but in lower proportions in P. hospiton. Introgression apparently occurred through past hybridization and repeated backcrossing, as evidenced by hybrid index scores and Bayesian assignment tests. Levels of introgression were low (0-1%) at two sex-linked loci and mitochondrial DNA, limited (0.4-2%) at three autosomal loci coding for dimeric enzymes, and high (up to 43%) at four autosomal loci coding for monomeric enzymes. Accordingly, selective filters are acting against foreign alleles, with differential effectiveness depending on the loci involved. The low levels of introgression at sex-linked loci and mitochondrial DNA are in agreement with Haldane's rule and suggest that introgression in P. machaon proceeds mainly through males, owing to a lower fitness of hybrid females. Papilio machaon populations showed higher levels of introgression in Sardinia than in Corsica. The role of reinforcement in the present reproductive isolation between P. machaon and P. hospiton is examined, as well as the evolutionary effects of introgressive hybridization between the two species. PMID- 12755876 TI - Phylogeography of Cavernularia hultenii: evidence of slow genetic drift in a widely disjunct lichen. AB - Population structure and history is poorly known in most lichenized ascomycetes. Many species display large-scale infraspecific disjunctions, which have been explained alternately by range fragmentation in species of high age and widespread long-distance dispersal. Using the lichen Cavernularia hultenii, which is widely disjunct across North America and Europe, Pleistocene and Holocene population history was inferred. The internal transcribed spacer (ITS) and part of the the intergenic spacer (IGS) region of the nuclear ribosomal DNA were sequenced in 300 individuals representing 62 populations across the range of the species. While four ancestral haplotypes are found in all areas, none of the observed tip haplotypes is present in more than one of the three part ranges. Although this is evidence for a past fragmentation event, nested clade analysis (NCA) remains equivocal in the choice between allopatric fragmentation and long distance dispersal. Mismatch distributions indicate exponential population growth, probably during postglacial invasion of C. hultenii into formerly glaciated areas of western North America. The presence of one southern and at least one northern glacial refugium in South Central Alaska is inferred. Evidence for another refugium in the Queen Charlotte Islands or Alexander Archipelago is inconclusive because of sparse sampling. However, a range expansion was not confirmed unambiguously by NCA. The limited power of NCA to infer past range fragmentations and expansions is due apparently to the shallow haplotype network and widespread ancestral haplotypes. This can be explained by slow genetic drift causing incomplete removal of ancestral haplotypes from the postfragmentation and postexpansion areas. PMID- 12755877 TI - Phylogeography of maritime pine inferred with organelle markers having contrasted inheritance. AB - Range-wide variation of maritime pine was studied at maternally inherited and paternally inherited markers (mitochondrial DNA and chloroplast DNA). While chloroplast DNA exhibits the highest diversity, phylogeographic inferences from this marker are blurred by homoplasy and extensive pollen flow. In contrast, the only three mitochondrial haplotypes found provide a clear picture of nonoverlapping areas colonized from different refugia, with no single population having a mixed composition (GST = 1). Comparison of the genetic structure inferred from both organelle genomes allows the investigation of differential seed and pollen dispersal, pointing to pollen, but not seed, dispersal across the Strait of Gibraltar (from Morocco into Iberia). A comparison with already available genetic information, especially that of one of the maritime pine's most threatening insect pests, the bast scale Matsucoccus feytaudi, further completes the picture. PMID- 12755878 TI - Evidence of a hybrid-zone in Atlantic cod (Gadus morhua) in the Baltic and the Danish Belt Sea revealed by individual admixture analysis. AB - The study of hybrid zones is central to our understanding of the genetic basis of reproductive isolation and speciation, yet very little is known about the extent and significance of hybrid zones in marine fishes. We examined the population structure of cod in the transition area between the North Sea and the Baltic Sea employing nine microsatellite loci. Genetic differentiation between the North Sea sample and the rest increased along a transect to the Baltic proper, with a large increase in level of differentiation occurring in the Western Baltic area. Our objective was to determine whether this pattern was caused purely by varying degrees of mechanical mixing of North Sea and Baltic Sea cod or by interbreeding and formation of a hybrid swarm. Simulation studies revealed that traditional Hardy-Weinberg analysis did not have sufficient power for detection of a Wahlund effect. However, using a model-based clustering method for individual admixture analysis, we were able to demonstrate the existence of intermediate genotypes in all samples from the transition area. Accordingly, our data were explained best by a model of a hybrid swarm flanked by pure nonadmixed populations in the North Sea and the Baltic Sea proper. Significant correlation of gene identities across loci (gametic phase disequilibrium) was found only in a sample from the Western Baltic, suggesting this area as the centre of the apparent hybrid zone. A hybrid zone for cod in the ecotone between the high-saline North Sea and the low-saline Baltic Sea is discussed in relation to its possible origin and maintenance, and in relation to a classical study of haemoglobin variation in cod from the Baltic Sea/Danish Belt Sea, suggesting mixing of two divergent populations without interbreeding. PMID- 12755880 TI - The influence of the Miocene Mediterranean desiccation on the geographical expansion and genetic variation of Androcymbium gramineum (Cav.) McBride (Colchicaceae). AB - Chloroplast DNA (cpDNA) restriction site and isozyme data were combined to explore the spatial-temporal influence of the Messinian desiccation in the Mediterranean on the disjunct distribution of Androcymbium gramineum in Almeria and Morocco (north and south of the straits of Gibraltar, respectively). Lack of evidence for different selective pressures, divergence time estimates based on the calibration of the isozyme molecular clock with the cpDNA data, the basal position of Almerian populations in the A. gramineum clade, and the much higher isozyme polymorphism in Almeria suggest that (i) only a southern European range of A. gramineum existed before the Messinian [ approximately 11.2 million years ago (Ma), in the middle Miocene] and (ii) the desiccation of the Mediterranean basin about 5.5-4.5 Ma induced the migration of A. gramineum from Almeria to Morocco (between 4.9 and 4.6 Ma, according to our time estimates). After the split into two allopatric units following the refilling of the Mediterranean, the major influence of drift associated with Plio-Pleistocene recurrent glaciation cycles and range expansions/contractions probably fostered the substantial interpopulation genetic differentiation observed within Almeria (CGST = 0.41, average DNei = 0.185) and, to a lesser extent, within Morocco (CGST = 0.24, average DNei = 0.089), but did not hinder the maintenance of considerable levels of genetic variation in either geographical area (A = 2.14, HE = 0.230 and A = 1.90, HE = 0.213, respectively). PMID- 12755879 TI - Molecular evidence reveals a polyphyletic origin and chromosomal speciation of Lake Baikal's endemic asellid isopods. AB - The six endemic isopod species of Lake Baikal have been regarded as a small species flock with uncertain affinities to related asellids. We provide evidence from 16S rRNA sequences for polyphyletic origins of Baikalian Asellidae. One clade of two species is related to the Eurasian genus Asellus. The other clade, Baicalasellus, shows affinities to North American asellids and may have a long evolutionary history within the lake basin. Some speciation events within Baicalasellus clearly have a chromosomal basis. In contrast with numerous taxa exhibiting monophyletic radiations in ancient lakes, the endemic Baikalian isopods arose by multiple invasions and chromosomal mechanisms. PMID- 12755881 TI - Genetic consequences of the ice ages on nurseries of the bat Myotis myotis: a mitochondrial and nuclear survey. AB - Analyses of mitochondrial DNA (mtDNA) control region polymorphism and of variation at 10 nuclear microsatellite loci were used to investigate the mechanisms and genetic consequences of postglacial expansion of Myotis myotis in Europe. Initial sampling consisted of 480 bats genotyped in 24 nursery colonies arranged along a transect of approximately 3000 km. The phylogeographical survey based on mtDNA sequences revealed the existence of major genetic subdivisions across this area, with several suture zones between haplogroups. Such zones of secondary contact were found in the Alps and Rhodopes, whereas other potential barriers to gene flow, like the Pyrenees, did not coincide with genetic discontinuities. Areas of population admixture increased locally the genetic diversity of colonies, which confounded the northward decrease in nucleotide diversity predicted using classical models of postglacial range expansion. However, when analyses were restricted to a subset of 15 nurseries originating from a single presumed glacial refugium, mtDNA polymorphism did indeed support a northwards decrease in diversity. Populations were also highly structured (PhiST = 0.384). Conversely, the same subset of colonies showed no significant latitudinal decrease in microsatellite diversity and much less population structure (FST = 0.010), but pairwise genetic differentiation at these nuclear markers was strongly correlated with increasing geographical distance. Together, this evidence suggests that alleles carried via male bats have maintained enough nuclear gene flow to counteract the effects of recurrent bottlenecks generally associated with recolonization processes. As females are highly philopatric, we argue that the maternally transmitted mtDNA marker better reflects the situation of past, historical gene flow, whereas current levels of gene flow are better reflected by microsatellite markers. PMID- 12755883 TI - Demographic expansion of parasitic nematodes of livestock based on mitochondrial DNA regions that conflict with the infinite-sites model. AB - Mitochondrial ND4 sequences of populations of four species of parasitic nematodes of livestock were subjected to demographic analyses. Deviation from selective neutrality was detectable using the frequency spectrum of segregating sites and highly negative neutrality statistics. However, the mitochondrial data sets do not comply with the infinite-sites model that underlies these tests, and as a consequence, it was not established whether these features are solely a result of population expansion, or whether aspects of the molecular evolution of these mitochondrial regions are also involved. Coalescent analyses based on Fu's Fs neutrality test, which incorporated estimates of rate heterogeneity, the transition-transversion ratio and nucleotide bias, as well as analyses that are fairly robust to deviations from the infinite-sites model supported population expansion. Also analyses that do not depend on the infinite-sites model suggested historical population expansion of these nematodes. The very similar time since expansion, the absence of signatures of positive selection in ND4 and the logical association with human demography imply that selective sweeps of mitochondrial variants are less probable, and that expansion is the most likely scenario for the parasitic nematodes of livestock. The methods used to characterize the expansion have different assumptions and emphasize different aspects of expansions. The resulting restrictions on the interpretation of expansions are outlined. PMID- 12755882 TI - Population genetic structure of the toad Bufo woodhousii: an empirical assessment of the effects of haplotype extinction on nested cladistic analysis. AB - Nested cladistic analysis (NCA) is increasingly being used to infer historical population-level processes, including population fragmentation, range expansion and long-distance colonization. However, the effects on interpretation of NCA inferences of stochastic extinction of haplotypes due to genetic drift (lineage sorting), or of haplotype loss via localized biotic or climatic influences, have not been thoroughly explored. We provide empirical evidence suggesting that NCA may misinterpret population history when haplotypes or haplotype groups from one clade are replaced by those of another clade. We do so by using NCA to analyse mitochondrial sequences from the toad Bufo woodhousii from 45 locations spanning the Great Plains and southwestern USA. Portions of this region were glaciated and/or desertified in the late Pleistocene and early Holocene, and hence uninhabitable for plains-dwelling organisms. Although NCA inferences of isolation by-distance and gradual range expansion in B. woodhousii are compatible with expectations based on climatic data and toad biology, NCA also detected several instances of long-distance movement. Such movement seems unlikely, given the low vagility of this species. We conclude that inferences of long-distance colonization likely result from extinction of haplotypes in intervening areas. We suggest using additional methods to look for congruent inferences, and amending the NCA inference key, to help avoid misinterpretations resulting from haplotype extinction. PMID- 12755884 TI - Early male reproductive advantage, multiple paternity and sperm storage in an amphibian aggregate breeder. AB - We tested whether the order in which males encounter females affects reproductive fitness in spotted salamanders (Ambystoma maculatum). Using mating chambers in the field, we allowed one male access to a female before a second male. We then used four microsatellite markers in paternity analyses of the resulting larvae. First males sired a significantly larger number of offspring than second males, suggesting that male reproductive success is greatly enhanced by early arrival at breeding ponds. Multiple paternity was common among clutches, and frequently larvae were assigned to unidentified males that had not been in the chambers. Sperm from these males had either been stored by females for a year or obtained more recently at other breeding sites. PMID- 12755885 TI - Estimation of pairwise relatedness between individuals and characterization of isolation-by-distance processes using dominant genetic markers. AB - A new estimator of the pairwise relatedness coefficient between individuals adapted to dominant genetic markers is developed. This estimator does not assume genotypes to be in Hardy-Weinberg proportions but requires a knowledge of the departure from these proportions (i.e. the inbreeding coefficient). Simulations show that the estimator provides accurate estimates, except for some particular types of individual pairs such as full-sibs, and performs better than a previously developed estimator. When comparing marker-based relatedness estimates with pedigree expectations, a new approach to account for the change of the reference population is developed and shown to perform satisfactorily. Simulations also illustrate that this new relatedness estimator can be used to characterize isolation by distance within populations, leading to essentially unbiased estimates of the neighbourhood size. In this context, the estimator appears fairly robust to moderate errors made on the assumed inbreeding coefficient. The analysis of real data sets suggests that dominant markers (random amplified polymorphic DNA, amplified fragment length polymorphism) may be as valuable as co-dominant markers (microsatellites) in studying microgeographic isolation-by-distance processes. It is argued that the estimators developed should find major applications, notably for conservation biology. PMID- 12755886 TI - Queen control over reproductive decisions--no sexual deception in the ant Lasius niger. AB - Queen-worker conflicts in social insect societies have received much attention in the past decade. In many species workers modify the colony sex ratio to their own advantage or produce their own male offspring. In some other species, however, queens seem to be able to prevent workers from making selfish reproductive decisions. So far, little effort has been made to find out how queens may keep control over sex ratio and male parentage. In this study we use a Lasius niger population under apparent queen control to show that sexual deception cannot explain queen dominance in this population. The sexual deception hypothesis postulates that queens should prevent workers from discriminating against males by disguising male brood as females. Contrary to the predictions of this hypothesis, we found that workers are able to distinguish male and female larvae early in their development: in early spring workers generally placed only either female or male larvae in the uppermost chambers of the nest, although both types of larvae must have been present. At this time males were only at 11% of their final dry weight, a developmental stage at which (according to two models) workers would still have benefited from replacing queen-produced males by females or worker-produced males. This study thus demonstrates that sexual deception cannot account for the apparent queen control over colony sex ratio and male parentage in L. niger. PMID- 12755887 TI - Queen-controlled sex ratios and worker reproduction in the bumble bee Bombus hypnorum, as revealed by microsatellites. AB - Social insect colonies provide model systems for the examination of conflicts among parties with different genetic interests. As such, they have provided the best tests of inclusive fitness theory. However, much remains unknown about in which party's favour such conflicts are resolved, partly as a result of the only recent advent of the molecular tools needed to examine the outcome of these conflicts. Two key conflicts in social insect colonies are over control of the reproductive sex ratio and the production of male offspring. Most studies have examined only one of these conflicts but in reality they occur in tandem and may influence each other. Using microsatellite analyses, the outcome of conflict over sex ratios and male production was examined in the bumble bee, Bombus hypnorum. The genotypes were determined for mother queens, their mates and males for each of 10 colonies. In contrast to other reports of mating frequency in this species, all of the queens were singly mated. The population sex ratio was consistent with queen control, suggesting that queens are winning this conflict. In contrast, workers produced over 20% of all males in queen-right colonies, suggesting that they are more effective in competing over male-production. Combining these results with previous work, it is suggested that worker reproduction is a labile trait that may well impose only small costs on queen fitness. PMID- 12755888 TI - Characterization of Pacific golden chanterelle (Cantharellus formosus) genet size using co-dominant microsatellite markers. AB - We characterized five co-dominant microsatellite markers and used them to study Pacific golden chanterelle (Cantharellus formosus) genet size and its relation to forest age and disturbance. Fruit-bodies were mapped in and collected from nine replicate study plots in old-growth, recently thinned, and unthinned 40-60-year old second-growth stands dominated by Douglas fir (Pseudotsuga menziesii). Information from microsatellite loci, combined with random fragment length polymorphism analysis of the nuclear DNA internal transcribed spacer indicates that putative 'C. formosus' fruit-body collections may include a cryptic chanterelle species. Small genets were characterized for both genetic types with mean maximum widths of 3.2 +/- 3.6 m for C. formosus and 1.5 +/- 1.7 m for the alternative genetic group. Variance in genet size was high and some multilocus genotypes were observed on multiple plots separated by 0.3 km or more, indicating that genets were not fully resolved by the loci described here. There was no evidence that genet size differed across the three disturbance treatments. PMID- 12755889 TI - Evidence for interspecies transmission of viruses in natural populations of filamentous fungi in the genus Cryphonectria. AB - Interspecies transmission is a significant evolutionary event that has allowed a variety of pathogens to invade new host species. We investigated interspecies transmission of viruses between the chestnut blight fungus, Cryphonectria parasitica, and a sympatric unidentified Cryphonectria species in Japan. Two isolates of Cryphonectria sp. were found to contain Cryphonectria hypovirus 1 (CHV-1), which has been typically found in C. parasitica. Three lines of evidence support the hypothesis of interspecies transmission of CHV-1. First, host species occur sympatrically and therefore have the opportunity to come into physical contact. Second, we transmitted CHV-1 between species experimentally in the laboratory. Third, phylogenetic analysis of 476 bp of the ORF B region of CHV-1 showed that sequences from Cryphonectria sp. were more closely related to those from C. parasitica than to each other. Local geographical subdivision of virus sequences from both host species argues against the alternative hypothesis of independent evolution of CHV-1 since speciation of their hosts. Based on these findings, we rule out the hypotheses that CHV-1 diverged from viruses in a common ancestor of the hosts, or that ancestral polymorphisms in CHV-1 persisted in the two host taxa. Estimating the direction and frequency of interspecies transmission in nature will require more extensive samples of CHV-1 from both host species. PMID- 12755890 TI - Checking the geographical origin of oak wood: molecular and statistical tools. AB - New methods for better identification of timber geographical origin would constitute an important technical element in the forest industry, for phytosanitary certification procedures or in the chain of custody developed for the certification of timber from sustainably managed forests. In the case of the European white oaks, a detailed reference map of chloroplast (cp) DNA variation across the range exists, and we propose here to use the strong geographical structure, characterized by a differentiation of western vs. eastern populations, for the purpose of oak wood traceability. We first developed cpDNA markers permitting the characterization of haplotype on degraded DNA obtained from wood samples. The techniques were subsequently validated by confirming the full correspondence between genotypes obtained from living tissues (buds) and from wood collected from the same individual oak. Finally, a statistical procedure was used to test if the haplotype composition of a lot of wood samples is consistent with its presumed geographical origin. Clearly, the technique cannot permit the unambiguous identification of wood products of unknown origin but can be used to check the conformity of genetic composition of wood samples with the region of alleged origin. This could lead to major applications not only in the forest industry but also in archaeology or in palaeobotany. PMID- 12755891 TI - Fluorescence in situ hybridization: a new method for determining primary sex ratio in ants. AB - The haplodiploid sex determining system in Hymenoptera, whereby males develop from haploid eggs and females from diploid eggs, allows females to control the primary sex ratio (the proportion of each sex at oviposition) in response to ecological and/or genetic conditions. Surprisingly, primary sex ratio adjustment by queens in eusocial Hymenoptera has been poorly studied, because of difficulties in sexing the eggs laid. Here, we show that fluorescence in situ hybridization (FISH) can be used to accurately determine the sex (haploid or diploid) of eggs, and hence the primary sex ratio, in ants. We first isolated the homologue coding sequences of the abdominal-A gene from 10 species of 8 subfamilies of Formicidae. Our data show that the nucleotide sequence of this gene is highly conserved among the different subfamilies. Second, we used a sequence of 4.5 kbp from this gene as a DNA probe for primary sex ratio determination by FISH. Our results show that this DNA probe hybridizes successfully with its complementary DNA sequence in all ant species tested, and allows reliable determination of the sex of eggs. Our proposed method should greatly facilitate empirical tests of primary sex ratio in ants. PMID- 12755892 TI - Reliable microsatellite genotyping of the Eurasian badger (Meles meles) using faecal DNA. AB - The potential link between badgers and bovine tuberculosis has made it vital to develop accurate techniques to census badgers. Here we investigate the potential of using genetic profiles obtained from faecal DNA as a basis for population size estimation. After trialling several methods we obtained a high amplification success rate (89%) by storing faeces in 70% ethanol and using the guanidine thiocyanate/silica method for extraction. Using 70% ethanol as a storage agent had the advantage of it being an antiseptic. In order to obtain reliable genotypes with fewer amplification reactions than the standard multiple-tubes approach, we devised a comparative approach in which genetic profiles were compared and replication directed at similar, but not identical, genotypes. This modified method achieved a reduction in polymerase chain reactions comparable with the maximum-likelihood model when just using reliability criteria, and was slightly better when using reliability criteria with the additional proviso that alleles must be observed twice to be considered reliable. Our comparative approach would be best suited for studies that include multiple faeces from each individual. We utilized our approach in a well-studied population of badgers from which individuals had been sampled and reliable genotypes obtained. In a study of 53 faeces sampled from three social groups over 10 days, we found that direct enumeration could not be used to estimate population size, but that the application of mark-recapture models has the potential to provide more accurate results. PMID- 12755893 TI - Genetic marker investigation of the source and impact of predation on a highly endangered species. AB - In September and October 2000, the remains of a number of apparently predated northern hairy-nosed wombats (Lasiorhinus krefftii) were discovered in Epping Forest National Park, the site of the only known population of this highly endangered species. Analysis of DNA recovered from six carcasses and a section of intestine found nearby was carried out using microsatellite and Y-specific primers. This identified seven individual wombats, the identity of three of which was inferred from a genotype database prepared from animals sampled during trapping programmes. Six victims were male and one female, suggesting that female biased predation rates are unlikely to be the cause of the current male-biased population sex ratio. DNA isolated from four canid faeces found in the vicinity revealed three distinct canid microsatellite genotypes with very high probabilities of belonging to dingoes (Canis familiaris dingo). A wombat genotype matching that of one of the dead individuals was identified from scats of two of the dingoes. In addition, two macropod microsatellites were amplified from two dingo scats. These observations provided vital information regarding predation on northern hairy-nosed wombats, and prompted the permanent exclusion of dingoes from the park by the erection of a dingo-proof fence. PMID- 12755894 TI - Microspatial genetic heterogeneity and gene flow in stray cats (Felis catus L.): a comparison of coat colour and microsatellite loci. AB - We analysed levels of genetic differentiation between nine local urban colonies of stray cats using eight coat colour and nine microsatellite loci. Both types of markers revealed a strong differentiation between colonies (FST = 0.15 and 0.09 for coat colour and microsatellite loci, respectively). Three coat colour loci showed extreme levels of genetic differentiation comparatively to other loci and are strongly suspected to be under divergent selective pressures. Microsatellite loci showed significant heterozygote deficiency within colonies (FIS = 0.14), suggesting that coat colour loci are not appropriate to investigate genetic structure at a fine scale because coat colour allele frequencies are based on Hardy-Weinberg equilibrium. The reported pattern conformed to that predicted from the social structuring of cat colonies: aggressive exclusion of immigrants, inbreeding and very low dispersal rate. PMID- 12755895 TI - Edaphic races and phylogenetic taxa in the Lasthenia californica complex (Asteraceae: Heliantheae): an hypothesis of parallel evolution. AB - Lasthenia californica sensu Ornduff consists of two races that differ in their flavonoid pigments and edaphic tolerances. Recent phylogenetic studies of Lasthenia have revealed that members of L. californica sensu Ornduff belong to two phylogenetic species. The relationship of the edaphic races to these new species and to each other is the focus of this study. Characterization of flavonoid profiles and phylogenetic placement of 33 populations demonstrates that races and phylogenetic taxa are not concordant, suggesting that one or both edaphic races evolved in parallel in the two clades. We hypothesize an edaphically linked ecological role for flavonoid differences that first revealed the existence of two races. PMID- 12755896 TI - Measuring gene flow from two birdsfoot trefoil (Lotus corniculatus) field trials using transgenes as tracer markers. AB - Genetic engineering is becoming a useful tool in the improvement of plants but concern has been expressed about the potential environmental risks of releasing genetically modified (GM) organisms into the environment. Attention has focused on pollen dispersal as a major issue in the risk assessment of transgenic crop plants. In this study, pollen-mediated dispersal of transgenes via cross fertilization was examined. Plants of Lotus corniculatus L. transformed with either the Escherichia coli asparagine synthetase gene asnA or the beta glucuronidase gene uidA, were used as the pollen donor. Nontransgenic plants belonging to the species L. corniculatus L., L. tenuis Waldst. and Kit. ex Willd, and L. pedunculatus Cav., were utilized as recipients. Two experimental fields were established in two areas of central Italy. Plants carrying the uidA gene were partially sterile, therefore only the asnA gene was used as a tracer marker. No transgene flow between L. corniculatus transformants and the nontransgenic L. tenuis and L. pedunculatus plants was detected. As regards nontransgenic L. corniculatus plants, in one location flow of asnA transgene was detected up to 18 m from the 1.8 m2 donor plot. In the other location, pollen dispersal occurred up to 120 m from the 14 m2 pollinating plot. PMID- 12755898 TI - Influence of endotoxin on the disposition kinetics and dosage regimens of oxytetracycline in calves. AB - The influence of endotoxin on the disposition kinetics of oxytetracycline (OTC) (10 mg/kg) was investigated in five healthy ruminating male crossbred calves. The serum concentration-time data of OTC before and after endotoxin challenge were best described by a two-compartment open model. Repeated administration of Escherichia coli endotoxin (1 microg/kg, i.v.) at an interval of 12 h up to 48 h produced a clear rise in the body temperature and an increase in the pulse and respiration rates. Endotoxin caused a significant reduction in mean transit time in tissue compartment (MTTT) (P < or = 0.05), mean residence time in the peripheral tissue compartment (MRTT) (P < or = 0.05), mean residence time in the body (MRTB) (P < or = 0.05), elimination half-life (t1/2lambda2) (P < or = 0.05) and distribution space in tissues (VT) (P < or = 0.01) and at steady-state (Vd(ss)) (P < or = 0.01). Endotoxin had no effect on the distribution clearance (ClD), systemic clearance (Cl) and distribution half-life of OTC, while the values of first order rate constant of transfer of drug from tissue to central compartment (K21) and the zero time intercept at terminal phase (C2) were significantly high. The drug dosage regimens to maintain serum OTC concentrations of 0.5, 1, 2, 4, 6 and 8 microg/mL were also determined in febrile and clinically healthy animals. PMID- 12755899 TI - Variable absorption of clavulanic acid after an oral dose of 25 mg/kg of Clavubactin and Synulox in healthy dogs. AB - The aims of this investigation were to calculate the pharmacokinetic parameters and to identify parameters, based on individual plasma concentration-time curves of amoxicillin and clavulanic acid in dogs, that may govern the observed differences in absorption of both drugs. The evaluation was based on the data from plasma concentration-time curves obtained following a single dose in an open, randomized, two-way crossover study involving 24 male Beagle dogs treated with two Amoxi-Clav formulations (A Clavubactin and B Synulox, each with 200/50 mg). Plasma amoxicillin and clavulanic acid concentrations were determined using validated bioassay methods. The half-life of elimination of amoxicillin was 1.5 h (t1/2 = 1.52 +/- 0.19 h, Cmax = 11.4 +/- 2.74 microg/mL), and that of clavulanic acid 0.76 h (t1/2 = 0.71 +/- 0.23 h, Cmax = 2.06 +/- 1.05 microg/mL). There was a fivefold variation in the AUCt of clavulanic acid for both formulations, while the AUCt of amoxicillin varied by a factor of 2. The mean ratio of the AUCt amoxicillin : clavulanic acid was 12.7 +/- 3.65 for formulation A and 11.8 +/- 5.22 for formulation B (P = 0.51). PMID- 12755900 TI - The effects of danofloxacin and tilmicosin on neutrophil function and lung consolidation in beef heifer calves with induced Pasteurella (Mannheimia) haemolytica pneumonia. AB - Pneumonia caused by Pasteurella (Mannheimia) haemolytica was induced in weaned beef heifer calves, approximately 6 months of age. Calves were treated at 20 h after challenge with therapeutic doses of danofloxacin or tilmicosin. Peripheral blood neutrophils were collected at 3, 24 and 48 h after treatment. The ex vivo effects on neutrophil function, neutrophil apoptosis, and hematological parameters were examined, as was the effect on percentage lung consolidation. Neutrophil function assays included random migration under agarose, cytochrome C reduction, iodination, Staphylococcus aureus ingestion, chemotaxis, and antibody dependent and antibody-independent cell-mediated cytotoxicity. Apoptosis was determined using a cell death detection kit. Killing was performed at 72 h after treatment. Statistical comparisons were made among the three groups of challenged treated animals: saline, danofloxacin, and tilmicosin. Comparisons were also made between nonchallenged nontreated animals (NCH) and challenged saline-treated animals. There were no significant differences for any of the neutrophil function assays or neutrophil apoptosis among the challenged-treated groups. This suggests that danofloxacin and tilmicosin have no clinically significant effects on neutrophil function or apoptosis. There were also no significant differences in percentage lung consolidation among the challenged-treated groups. Significant differences were found between the NCH calves and the challenged saline-treated calves in several neutrophil assays, which were attributed to effects of P. haemolytica infection. PMID- 12755901 TI - The efficacy of a single intraperitoneal injection of oxolinic acid in the treatment of bacterial infections in goldsinny wrasse (Ctenolabrus rupestris) and corkwing wrasse (Symphodus melops) studied under field and laboratory conditions. AB - The efficacy of a single intraperitoneal injection of oxolinic acid to control an outbreak of atypical Aeromonas salmonicida infection in goldsinny wrasse (Ctenolabrus rupestris) and in the treatment of systemic vibriosis in corkwing wrasse (Symphodus melops) was examined. In addition a field study was performed to examine the effect of medication on the survival rate of goldsinny wrasse in Atlantic salmon cages. Four groups of wild caught goldsinny wrasse, each of 50 fish, were treated with an intraperitoneal injection of propylene glycol:saline (50:50) (control) or 50 mg/kg oxolinic acid at a concentration of 50 mg/mL. Three days after medication the fish in all groups were treated by an intraperitoneal injection of prednisolone acetate and an increase in seawater temperature from 9.0 to 11.5 degrees C. Cumulative mortalities were 18% in the two groups treated with oxolinic acid and 94 and 100% in the unmedicated control groups, giving a 'relative percentage survival' (RPS) value of 82%. A laboratory maintained population of originally wild caught corkwing wrasse experiencing high daily mortality was treated with oxolinic acid (50 mg/kg) or propylene glycol:saline (control). Cumulative mortalities were 84% (control) and 42% (oxolinic acid medicated group) giving an RPS value of 50%. In a field investigation using goldsinny wrasse approximately 30% were medicated with oxolinic acid (50 mg/kg) prior to stocking in cages with Atlantic salmon. In two of three cages the cumulative mortality was significantly lower (P = 0.025 and P < 0.001) in the medicated groups. PMID- 12755902 TI - The steady-state pharmacokinetics and bioequivalence of carprofen administered orally and subcutaneously in dogs. AB - Eighteen male Beagle dogs were randomized to oral (p.o.) or subcutaneous (s.c.) carprofen administration in a two-sequence, two-period crossover design with a 10 day washout between periods. Twenty-five milligrams of carprofen was administered p.o. or s.c. every 12 h for 7 days. Plasma concentrations of carprofen collected after the first and last treatments were determined by high-performance liquid chromatography. Carprofen concentration data were natural log transformed and geometric means were calculated for maximum plasma concentration (Cmax) and area under the plasma concentration-time curve (AUC0--12) following the first dose and Cmax and AUC0--12 following administration of the last dose. Formulations were considered bioequivalent if the 90% confidence interval (CI) of the mean difference for each variable between formulations were within -20% and 25% of the oral formulation. The mean Cmax and AUC0--12 were 16.9 microg/mL and 73.1 microg. h/mL, respectively, following a single oral dose and 8.0 microg/mL and 64.3 microg x h/mL, respectively, following a single s.c. injection. The 90% CI for Cmax (-56.8 to -48.7%) was outside of the bioequivalence criteria whereas the 90% CI for AUC0--12 (-16.3 to -7.5%) was within the bioequivalence criteria. At steady-state, the mean Cmax and AUC0--12 were 18.7 microg/mL and 101.9 microg x h/mL, respectively, following p.o. administration and 14.7 microg/mL and 111.0 microg x h/mL, respectively, following s.c. injection. The 90% CI was outside the bioequivalence criteria for Cmax (-30.8 to -10.8) but within the bioequivalence criteria for AUC0--12 (2.3-15.9%). The results of this study indicate that peak plasma concentrations of carprofen differ when administered p.o. and s.c., but that total drug exposure following a single dose and at steady-state are bioequivalent. PMID- 12755903 TI - Efficacy of intramammary treatment with procaine penicillin G vs. procaine penicillin G plus neomycin in bovine clinical mastitis caused by penicillin susceptible, gram-positive bacteria--a double blind field study. AB - The efficacy of intramammary treatments containing procaine penicillin G alone (treatment A) or a combination of procaine penicillin G and neomycin (treatment B) was compared in treating clinical bovine mastitis caused by gram-positive bacteria susceptible in vitro to penicillin G. Both treatments were supplemented with a single intramuscular injection of procaine penicillin G on the first day of treatment. The study was carried out using a double blind design on commercial dairy farms in Southern Finland. A total of 56 quarters were treated with treatment A and 61 with treatment B. The cure rates for both treatments were equal, which suggests that the use of the penicillin G-aminoglycoside combination does not increase the efficacy of the treatment over that achieved by using penicillin G alone in bovine clinical mastitis caused by penicillin-susceptible, gram-positive bacteria. PMID- 12755904 TI - A comparison of propofol infusion and propofol/isoflurane anaesthesia in dexmedetomidine premedicated dogs. AB - The effects of propofol infusion were compared with propofol/isoflurane anaesthesia in six beagles premedicated with 10 microg/kg intramuscular (i.m.) dexmedetomidine. The suitability of a cold pressor test (CPT) as a stress stimulus in dogs was also studied. Each dog received isoflurane (end tidal 1.0%, induction with propofol) with and without CPT; propofol (200 microg/kg/min, induction with propofol) with and without CPT; premedication alone with and without CPT in a randomized block study in six separate sessions. Heart rate and arterial blood pressures and gases were monitored. Plasma catecholamine, beta endorphin and cortisol concentrations were measured. Recovery profile was observed. Blood pressures stayed within normal reference range but the dogs were bradycardic (mean heart rate < 70 bpm). PaCO2 concentration during anaesthesia was higher in the propofol group (mean > 57 mmHg) when compared with isoflurane (mean < 52 mmHg). Recovery times were longer with propofol than when compared with the other treatments. The mean extubation times were 8 +/- 3.4 and 23 +/- 6.3 min after propofol/isoflurane and propofol anaesthesia, respectively. The endocrine stress response was similar in all treatments except for lower adrenaline level after propofol infusion at the end of the recovery period. Cold pressor test produced variable responses and was not a reliable stress stimulus in the present study. Propofol/isoflurane anaesthesia was considered more useful than propofol infusion because of milder degree of respiratory depression and faster recovery. PMID- 12755905 TI - Interactions of xylazine and detomidine with alpha2-adrenoceptors in brain tissue from cattle, swine and rats. AB - Xylazine is an alpha2-adrenoceptor agonist sedative with a much higher interspecies variability in effect than detomidine, another alpha2-agonist used in veterinary practice. In the present study, we have used radioligand binding in brain tissue to investigate if the high species variation in sensitivity to xylazine could be explained in terms of receptor interactions. Species known to be more (cattle) or less (swine and rats) sensitive to xylazine were used. There was no variation in the density or the subtype pattern of the alpha2 adrenoceptors that could explain the species variation recorded in vivo, as a homogenous population of the alpha2A/D-subtype (200-300 fmol/mg protein) was found in all species. The species differences in the affinities of xylazine and detomidine were minor and similar for the two drugs. The only parameter investigated where a significant species difference was found for xylazine but not for detomidine was the slope of the inhibition binding curve when the G protein coupling was diminished. For xylazine this slope was considerably lower than unity (i.e. 0.77 +/- 0.075) using cattle preparations compared with 0.92 +/- 0.037 (mean +/- SE) and 0.90 +/- 0.028, respectively for swine and rats, while for detomidine this parameter was close to unity in all species (cattle, swine, rat). This finding indicates that the species variation in effect for xylazine could be due to differences at the G-protein level or further down-stream in the effect cascade. PMID- 12755906 TI - Pharmacokinetic/pharmacodynamic modelling of the disposition and effect of benazepril and benazeprilat in cats. AB - The disposition and effect of benazepril and its active metabolite, benazeprilat, were evaluated in cats using a pharmacokinetic/pharmacodynamic model. Cats received single 1 mg/kg doses of intravenous 14C-benazeprilat and oral 14C benazepril.HCl, and single and repeat (eight daily) oral administrations of 0.25, 0.5 and 1.0 mg/kg nonlabelled benazepril.HCl. The pharmacokinetic endpoints were plasma concentrations of benazepril and benazeprilat, and recovery of radioactivity in faeces and urine. The pharmacodynamic endpoint was plasma angiotensin-converting enzyme (ACE) activity. Benazeprilat data were fitted to an equation corresponding to a single-compartment model with a volume equal to the blood space (Vc = 0.093 L/kg). Within this space, benazeprilat was bound nonlinearly to ACE, which was mainly tissular (89.4%) rather than circulating (10.6%). Free benazeprilat was eliminated quickly from the central compartment (t1/2 approximately 1.0 h; Cl approximately 0.125 L/kg/h), elimination being principally biliary ( approximately 85%) rather than urinary ( approximately 15%). Nevertheless, inhibition of ACE was long-lasting (t1/2 16-23 h) due to high affinity binding of benazeprilat to ACE (Kd approximately 3.5 mmol/L, IC50 approximately 4.3 mmol/L). Simulations using the model predict a lack of proportionality between dose of benazepril, plasma benazeprilat concentrations and effect due to the nonlinear binding of benazeprilat to ACE. For example, increasing the dose of benazepril (e.g. above 0.125 mg/kg q24 h) produced only small incremental inhibition of ACE (either peak effect or duration of action). PMID- 12755908 TI - Pyriproxyfen concentration in the coat of cats and dogs after topical treatment with a 1.0% w/v spot-on formulation. PMID- 12755907 TI - Levamisole mucosal adjuvant activity for a live attenuated Escherichia coli oral vaccine in weaned pigs. AB - The present study tested the hypothesis that levamisole exerts its immunopotentiating activity in weaned pigs vaccinated against colibacillosis by priming the lymphocytes and macrophages in the mesenteric lymph node (MLN). Ten weaned piglets were used and allocated into two equal groups. The experimental group was intramuscularly primed with levamisole at an immunostimulatory dose of 2.5 mg/kg given daily, in three consecutive days, and controls received saline according to the same schedule. Both groups were orally vaccinated with the vaccinal Escherichia coli strain on day 0 and challenged with the virulent E. coli strain 7 days later. All pigs were killed on postchallenge day 6. Upon virulent challenge the health status of the two groups was evaluated by clinical observations, and expression of CD25, SWC7 and SWC9 activation antigens by MLN and spleen T and B cells and macrophages, respectively, was tested using flow cytometry. Priming by levamisole significantly contributed to the effectiveness of a live attenuated oral vaccine against porcine postweaning colibacillosis, as evidenced by a good health status of primed vaccinated vs. un-primed vaccinated pigs. The CD3+, CD25+ and SWC9+ MLN but not spleen T cells and macrophages increased in experimental vs. control pigs, implying that levamisole exerts its potentiating activity in the MLN by augmenting both recruitment and activation of cells that participate in cell-mediated immunity. PMID- 12755909 TI - The differences and commonalities between United Kingdom and Canadian Psychiatric/Mental Health nursing: a personal reflection. PMID- 12755910 TI - Becoming a success story: how boys who have molested children talk about treatment. AB - This grounded theory study was designed to generate a mid-range theory of treatment from the perspective of boys who have molested children and undergone outpatient treatment. Data included information from seven boys' charts, their written responses to open-ended questions, and audio-taped interviews. The interviews generated a series of statements reflecting the boys' experiences in treatment, which were analysed by the constant comparative method. The basic social process of treatment was 'becoming a success story'. The structural elements of becoming a success story included relapse prevention, compliance and decision-making. The boys integrated these structural elements by talking to people they trusted, listening to what people said, and using what people said to help them do what was right. Becoming a success story took place in a context of family and community support. In its current form, this theory of treatment success can be used in practice to monitor progress through treatment. PMID- 12755911 TI - Role changes experienced by clinical staff in relation to clients' recovery from psychosis. AB - This study investigates the subjective experiences of staff from many interdisciplinary teams working with clients in the recovery process from psychosis. The clinical staff interviewed in this study included: staff nurses, clinical nurse specialists, occupational therapists, psychologists, social workers, rehabilitation workers, recreation therapists, music therapists, psychiatrists, and lodging home operators. The purpose of this study was to examine the clinical staff's changing roles and relationships with clients recovering from psychosis. The investigation used a naturalistic qualitative design with an ethnographic method of data analysis. The participants were clinical staff working with clients about to commence treatment with clozapine or risperidone. The settings used were a tertiary-care psychiatric hospital and a general hospital. The clinical staff members who participated in the study were interviewed every 3 months. In the initial interview, members of the clinical staff were asked about their knowledge regarding the new medication and their role in the decision to try the new medication. In all the interviews, clinical staff members were asked about how the recovery process was progressing with the client. Data regarding clinical staff fears related to the client's situation, changes in their relationships with the client and the client's family, and what they perceived to be current rehabilitation implications, were collected. Some of the clinical staff roles that evolved during the recovery process were: health teacher; advocate; counsellor; and support person. PMID- 12755912 TI - Impact of caregiving: listening to the voice of informal caregivers. AB - Long-term care of the elderly and the disabled rests on a vast network of informal caregivers. This qualitative study examined the effects of caregiving on the health and wellbeing of a small, non-representative sample of urban women caregivers. These 11 informants participated in two interviews. Health was conceptualized in a holistic manner, consistent with the view of the World Health Organization and Health Canada. The caregiving experience, as described in this study, was life changing and consuming. Analysis revealed that the domain of caregiving could be captured by three dominant themes: loss and grieving; adapting and coping; and the short- and long-term impact. The consistency in the findings and policy recommendations of studies of informal caregiving begs the question: when will caregivers finally be heard? PMID- 12755913 TI - Risk assessment and management of patients with self-neglect: a 'grey area' for mental health workers. AB - This study explores the perceptions and experiences of community mental health workers who assess and manage the risk of self-neglect and severe self-neglect in people with serious mental health problems. The initial literature review demonstrated a lack of material on this specific subject. Semi-structured interviews were undertaken with seven community mental health workers who met the criteria of being knowledgeable, skilled and credible practitioners. Data analysis was completed using thematic content analysis. Findings suggest that community mental health workers are operating in a number of areas where there is a distinct lack of clarity--'The Grey Areas'. The most important of these were the tolerance of workers to situations of self-neglect/severe self-neglect, policies, procedures and legislation, and definitions of self-neglect/severe self neglect. The workers dealt with this lack of clarity by completing a detailed risk assessment, which then enabled them to manage the risk. However, the need to balance the safety needs of clients against their need to be treated as autonomous individuals presented a major dilemma for workers. PMID- 12755914 TI - 'One flew over the psychiatric unit': mental illness and the media. AB - Media representation of mental illness has received growing research attention within a variety of academic disciplines. Cultural and media studies have often dominated in this research and discussion. More recently healthcare professionals have become interested in this debate, yet despite the importance of this subject only a selection of papers have been published in professional journals relating to nursing and healthcare. This paper examines the way in which mental illness in the United Kingdom is portrayed in public life. Literature from the field of media studies is explored alongside the available material from the field of mental healthcare. Three main areas are used to put forward an alternative approach: film representation and newspaper reporting of mental illness; the nature of the audience; and finally the concept of myth. The paper concludes by considering this approach in the context of current mental health policy on mental health promotion. PMID- 12755915 TI - Interventions with family caregivers for people with dementia: efficacy problems and potential solution. AB - Family members are increasingly caring for relatives with dementia at home. It is well recognized that this is a stressful experience for carers. Treatment approaches that aim to alleviate this stress were introduced in the 1970s and later research began to evaluate their efficacy. Early studies using subjective ratings of satisfaction and usefulness demonstrated encouraging results. However, later studies using standardized measures of distress and burden, have not been able to demonstrate efficacy in a convincing way. There have been some positive significant changes; however, most findings have been poor, inconsistent and equivocal. This may be accounted for by the poor methodological rigour diluting the positive potential benefits of interventions for carers. This paper suggests that in order for the efficacy research to evolve, there needs to be a change in direction. It is vital that instead of repeating 'more of the same' studies, researchers need to prioritize the use of theoretically driven interventions and research design. This factor alone could provide the framework to influence the methodological issues that potentially dilute the demonstration of treatment efficacy. Without clearer evidence from research, guidance for practitioners about treatment for carers is ambiguous. PMID- 12755917 TI - Manipulation: description, identification and ambiguity. AB - The word manipulation is frequently applied to some of the difficult-to-manage behaviours of the personality-disordered patient. However, the term is rarely defined, and a review of both the clinical and research literature shows that little has been written about its definition and identification, let alone its clinical management in both in- and outpatient settings. Recent empirical work conducted with nurses in forensic settings has demonstrated the range of behaviours that professionals refer to as 'manipulative', thus clarifying the use of the term and allowing the provision of a more precise definition. The scope of manipulation in everyday life, management practice and politics is perhaps relatively small, although manipulation can occur in all areas of human activity. Social behaviour is doubly ambiguous with respect to judgements of manipulation, as such judgements involve a moral evaluation combined with the identification of deception on the basis of little or partial evidence. The implications of this social ambiguity for clinical psychiatric practice are that professionals need to guard themselves from two polar faults: seeing manipulation everywhere; or being blind to its presence. In order to achieve a cautious moderation, staff need to hold both alternatives in mind at all times. PMID- 12755916 TI - Mutuality as background music in women's lived experience of mental health and depression. AB - Mental health problems, especially depression, have turned into an extensive public health problem, affecting women in particular. The aim of this study was to obtain a deeper understanding of mental health phenomena through elderly women's lived experiences of mental health and depression. The phenomenological approach was chosen for collecting experience-based and person-centred descriptions from 21 women, focusing on mental health and depression. The interviews were analysed using Giorgi's phenomenological descriptive method. Mutuality in their relationships with themselves and others emerged as a major element in the women's experience of mental health and depression. When the women's existence and value were confirmed in relation to themselves and others, mental health appeared as an ascending spiral. When the women's value and self esteem in relation to themselves and others were violated, the result was a descending spiral. Metaphorically speaking, 'mutuality' provided the 'background music' to the women's lives. Being a skilled professional psychiatric nurse means making the best use of mutuality as a creative power in the nurse-patient relationship. This means that the nurse must be aware that her/his attitude, appearance and behaviour are interpreted as a confirmation of the patient's worthiness or worthlessness. PMID- 12755918 TI - Manipulation: searching for an understanding. AB - Manipulative behaviour is one of the reasons why some psychiatric professionals dislike personality-disordered (PD) patients and dislike working with them. Being manipulated arouses strong negative emotions towards the manipulator and can have severe repercussions on the ability to care for or treat those suffering from personality disorders. In order for practitioners to cope with manipulative behaviour and manage their emotional responses towards it, it is necessary to find ways to understand or construe that behaviour, so that moral condemnation and rejection of the patient can be avoided. This article offers a summary of three such methods of interpreting manipulation which are to be found in the literature: manipulation as normal behaviour; manipulation as unconsciously motivated; and manipulation as cognitive distortion. These three viable schemes for the understanding of manipulation provide not just a means to enable a non judgmental approach, they also provide ways to manage that behaviour in a therapeutic manner. PMID- 12755919 TI - The meaning of respite care to mothers of children with learning disabilities: two Irish case studies. AB - There is a growing interest in Ireland in the nature and significance of respite care for carers and those for whom they care. The relationship of individual stress with caring full time for a child who is learning disabled is well documented. Provision of respite care is seen as an important means of alleviating individual carer stress. Yet, the apparent benefits of respite care have been called into question. The present study looks at this issue within the context of respite service provision in Ireland for young people with learning disabilities. A phenomenological approach was used to explore the views of two mothers on respite care and, in particular, its personal significance for them within the context of their caring relationship for their children. The authors found that for these two mothers, whilst some of the predicted benefits of respite care were present, for example improved social activity, their use of respite care and the experience of separation initiated feelings of guilt and appeared to engender a degree of emotional stress. It is argued that providers of respite services in Ireland need to consider how they can support parents who use respite care so that they see its use as a mark of caring for their child and thereby alleviate such feelings of guilt. PMID- 12755920 TI - The role of a mental health consumer in the education of postgraduate psychiatric nursing students: the students' evaluation. AB - Recent Australian Government policy reflects the integral nature of active consumer participation to the planning and delivery of mental health services. The effectiveness of consumer participation in improving mental health services has received some attention in the literature. Commonwealth Government funding enabled the development of a partnership between the Centre for Psychiatric Nursing Research and Practice and the Melbourne Consumer Consultants' Group. The successful application enabled the employment of a mental health consumer as an academic staff member of the Centre for Psychiatric Nursing Research and Practice. One important aspect of this role involved the mental health consumer teaching a consumer perspective to postgraduate psychiatric nursing students. The primary aim was to increase the students' awareness of and sensitivity to greater consumer participation within the mental health arena. This paper presents the results of an evaluation of the consumer academic role in teaching within the Postgraduate Diploma in Advanced Clinical Nursing (Psychiatric Nursing). An evaluation form was distributed to students (n = 21) on completion of the semester. The findings suggest the experience was considered beneficial to students and was impacting significantly on their current practice. This project supports the value of consumer participation in the education of mental health professionals. PMID- 12755922 TI - Perspectives of mindfulness. AB - The idea and practice of mindfulness has a long history in some of the world's religions and is also articulated in a number of secular discourses. The therapeutic potential of mindfulness is now being recognized and is being researched in a diverse range of healthcare settings including mental health. Being mindful presupposes that individuals whose awareness is not impaired do have a choice in what phenomena they attend to and how they act. For the psychiatric nurse, understanding the idea and practice of mindfulness is useful for developing both trans-cultural awareness and to recognize that personal inattention could compromise care. In terms of mental health promotion, mindfulness is worthy of consideration as an important life skill. This paper explores the notion of mindfulness from a number of perspectives. The paper does not claim to have explored all the options. The Buddhist understanding of mindfulness provides the perspective for continuity within the paper and is used to illuminate any similarities and differences with the secular discourses being considered. PMID- 12755921 TI - Patient satisfaction with psychiatric services provided by a Melbourne tertiary hospital emergency department. AB - The mainstreaming of psychiatric services within the general healthcare system has created fundamental changes to the manner in which patients access acute psychiatric services. This change was intended to reduce the stigma associated with psychiatric diagnosis and therefore contribute to improved treatment outcomes for patients. The aim of this paper is to discuss the results of a study designed to ascertain the level of psychiatric patient satisfaction with the services received in the emergency department of a Melbourne metropolitan hospital. The results indicate a high level of satisfaction, particularly with the availability of staff with psychiatric qualifications and experience to provident treatment, support and care. The major areas of dissatisfaction identified by patients included: lengthy waiting times, lack of privacy in the triage area and negative attitudes of general staff. These findings support the argument from the literature for psychiatric consultancy services to be available in the emergency department, and further identifies the need for triage guidelines to be tailored to the needs of mental health patients and for emergency department triage staff to be appropriately educated to adequately triage these patients. PMID- 12755923 TI - Palliative care within secure forensic environments. AB - The principles and philosophy of palliative care can be implemented successfully in secure forensic environments. While incarcerated, many inmates will suffer from a variety of life-limiting illnesses requiring palliative care. There are a number of resources available for the development and implementation of programs within secure forensic environments. PMID- 12755924 TI - A systematic review of the effectiveness of stress-management interventions for mental health professionals. PMID- 12755925 TI - Client and witness: the provision of therapy for vulnerable or intimidated adult witnesses prior to a criminal trial: practice guidance. PMID- 12755926 TI - Schizophrenia: the 'not-so-nice' guidelines. PMID- 12755927 TI - Poverty, underdevelopment and infant mental health. AB - Very great advances have occurred in disciplinary and professional knowledge of infant development and its influence on subsequent development. This expertise includes the ways in which early experiences affect the capacity of mature individuals for social adjustment and productive competence, and promising methods of intervention to promote infant mental health and prevent adverse sequelae of risk conditions. However, very little of this knowledge has been applied in work among infants and children living in conditions of poverty and underdevelopment. This lack of application continues despite the enormous threats to the well-being of infants and young children brought about by the combined effects of poverty and the AIDS pandemic, especially in southern Africa. Protein energy malnutrition, maternal depression, and institutional care of infants and small children are cited as illustrative of areas in which interventions, and their evaluation, are desperately needed in resource-poor countries. An argument is made for the critical importance of considering and addressing psychological factors in care givers and children in conditions of extreme material need. An example is provided of a simple intervention model based on sound developmental principles that can be implemented by trained non-professionals in conditions of poverty and underdevelopment. PMID- 12755928 TI - Parents, infants and health care: utilization of health services in the first 12 months of life. AB - OBJECTIVE: To describe patterns of health-service use in the first 12 months of life. METHODS: In this prospective cohort study, 173 first-born infants and their families living in two middle socio-economic urban areas of Melbourne were enrolled consecutively when presenting for their initial maternal and child health nurse (MCHN) visit (at approximately 4 weeks of age). Families kept a daily "health diary" for the entire 12-month period, recording use of all health services for their infant, and reasons for the contact. RESULTS: There was an 87% completion rate of diaries. The mean number of visits to any health service, including medical, hospitals, MCHN services, pharmacists, allied health services and naturopaths, was 35.7 (95% CI 34.7-36.6) during the 12 months. Of these, 31% (mean 10.9 visits) were visits to a general practitioner (GP) and 41.5% (mean 14.3 visits) were visits to the MCHN. Infants' visits to the MCHN were far more frequent in the first 6 months of life compared with the second 6 months (10.3 vs 3.6, P < 0.001). Rates of GP use were constant over the same periods (5.3 vs 5.7, P = 0.8). CONCLUSIONS: In a universal health-care system, this high rate of health-service use equates to approximately one visit to a health service every 2 weeks in the first year of life. The majority of these visits appeared unrelated to illness. This previously undocumented data has implications for future integrated service delivery, health-professional training and policy development for this age group. PMID- 12755929 TI - Economic effects of childhood cancer on families. AB - OBJECTIVE: To assess the financial impact of childhood cancer on families. METHODS: This was a cross-sectional survey of parents caring for children who were diagnosed with cancer during the period 1990-1993. Self-administered questionnaires were completed by the parents of 237 children from throughout New Zealand with different types of cancer. Dollar amounts were adjusted to the equivalent of December 2000. RESULTS: Eighty-six per cent of the 192 living children were well or in remission. A further 45 children had died. The average extra amount spent, because of the child's illness, by the family of a living child in the 30 days prior to participation in the study was NZ$220 (SD NZ$330). On average, this was 13% of the family income after tax. After reported entitlement to compensation from various sources was allowed for, families were left with a mean deficit of NZ$157 (SD NZ$278) for the 30 days. Twelve families had a shortfall of more than NZ$500, including three families that had a shortfall of more than NZ$1000. Expenditure was greater for those whose children spent more time in hospital (P = 0.003). There was no significant association between the total cost and the distance travelled to the treatment centre (P = 0.96). For 24 families, after-tax income in the month prior to participation in the study was at least NZ$500 lower than it had been in the month before the child's diagnosis. Thirty-seven per cent of families reported that they needed to borrow money because of the financial effects of the child's illness. Bereaved parents spent an average of NZ$3065 (SD NZ$2168) on funeral expenses. CONCLUSION: There is a large financial burden on families who have a child with cancer. PMID- 12755930 TI - Barriers to the use of interpreters in emergency room paediatric consultations. AB - OBJECTIVE: To identify barriers to the use of trained interpreters in emergency department (ED) paediatric consultations. METHODS: A cross-sectional survey of non-English-speaking-background (NESB) carers who presented a child aged 14 years or less to the Fairfield Hospital ED during a 3-month period was undertaken using a multilingual postal questionnaire or a phone interview with interpreter assistance. RESULTS: Of the 1388 paediatric consultations during the study period, 406 of the carers were registered as being of NESB, with 316 being eligible for study inclusion. Of the 278 (88%) respondents who completed the questionnaires, assistance with interpretation was required by 131 (47%) respondents: 55 (20%) used ad hoc interpreters, 47 (17%) used a trained interpreter and 21 (8%) did not receive the required assistance. Fifty-four (41%) carers who required interpreter assistance were identified as having this requirement at clerical registration in the ED, but only 18 of these were provided with a trained interpreter during the consultation. Limited English proficiency was self-reported by 127 (46%) respondents, with these respondents being more likely to need interpreter assistance (odds ratio (OR) 44.2; 95% CI 21.6-90.7), to have impaired understanding of the consultation (OR 8.2; 95% CI 4.7-14.1) and to consult a language concordant general practitioner (OR 10.1; 95% CI 5.1-19.4) compared with English proficient respondents. CONCLUSIONS: Barriers to the use of interpreters in ED paediatric consultations include poor identification of the need for and provision of an interpreter in the ED, and a preference for NESB carers to use ad hoc interpreters or no interpreter. Recommendations include the implementation in the ED of strategies to improve identification of NESB carers, as well as to improve awareness, access and use of trained interpreters during paediatric consultations. There is also a need to explore the experience of ED staff in accessing and using trained interpreters. PMID- 12755931 TI - Unintentional ingestion of over the counter medications in children less than 5 years old. AB - OBJECTIVE: Childhood ingestion of medications remains a substantial problem. Medication available over the counter (OTC) is widely used and has significant toxicity. The present study aims to investigate the nature and extent of unintentional ingestion of OTC medication in children < 5 years old in Victoria, Australia, during the period 1996-2000, in order to highlight critical factors. METHODS: Numbers of enquiries relating to unintentional ingestion of OTC medication in children < 5 years old and medication types were obtained from the Victorian Poisons Information Centre for 1998-2000. Emergency Department presentations involving poisoning of children < 5 years old, the medication types and subsequent admissions were obtained from the Victorian Emergency Minimum Dataset for 1996-2000. RESULTS: Numbers of enquiries and Emergency Department attendances for poisoning were substantially higher for OTC medication than for prescription medication; however, a lower proportion of cases involving ingestion of OTC medication (24.8%) required hospital admission during the study period compared with cases involving ingestion of prescription medications (33.8%). Overall, the peak incidence was at 2 years of age, with a slight male over representation. Paracetamol and cough/cold preparations were the most common agents. CONCLUSIONS: The causes of unintentional ingestion of OTC medications by children might include lack of child-resistant closure (CRC), inadequate design of CRC, attitudes concerning the toxicity of OTC medications, or lack of vigilance by parents and carers in the storage and administration of OTC medications. Consideration should be given to restricting sales of toxic OTC medications to pharmacies, and increasing counselling of consumers concerning the toxicity and safe storage of OTC medications and the correct usage of CRC. The adequacy of CRC design and OTC medications warranting CRC should be reviewed by the relevant authorities. PMID- 12755932 TI - Effect of detergent-coated versus non-coated spacers on bronchodilator response in children with asthma. AB - OBJECTIVE: Previous studies have demonstrated that coating spacers with ionic detergents minimizes the static charge and thereby improves in vivo drug deposition. The present study aims to examine the effect of coated spacers versus non-coated spacers in the clinical situation. METHODS: A randomized, double-blind study in children with asthma and a ratio of forced expiratory volume in 1 s to forced vital capacity (FEV1/FVC) of < or =72% predicted was carried out. Spirometry was performed at baseline and at 10 min and 20 min after inhalation of two puffs of salbutamol (100 microg/puff) through either a detergent-coated or a non-coated spacer. RESULTS: Fifty children were studied (mean age 11.6 years, range 7-18 years): 26 in the group using coated spacers (CG); and 24 in the group using non-coated spacers (NCG). The mean percentage change in FEV1 from baseline 10 min after inhalation was 18.8% (range 5-50%) in the CG versus 18.5% (range 3 35%) in the NCG. At 20 min after inhalation, the per cent increase in FEV1 was 19.8% (range 0-50%) in the CG versus 19.5% (range 9-35%) in the NCG. There was no significant difference between groups in the percentage change in FEV1 after 10 min (P = 0.91), or after 20 min (P = 0.93). CONCLUSIONS: There was no improvement in bronchodilatation from detergent-coated spacers in the present study, possibly because a maximal bronchodilator response was achieved with the lower output. PMID- 12755933 TI - Early manifestation and recognition of C2 complement deficiency in the form of pyogenic infection in infancy. AB - OBJECTIVE: Although frequently asymptomatic, C2 complement component deficiency may lead to severe pyogenic infections or lupus-like illness. In the present report, we describe infectious manifestations in infancy and childhood in our C2 deficient patients. METHOD: A retrospective study of clinical manifestation in three patients was carried out. C2 deficiency was proved both by undetectable serum C2 level and typical homozygous 28 bp deletion of the C2 gene. RESULTS: All patients were hospitalized at least once by the age of 12 months, each had one episode of meningitis in infancy, one also had arthritis with septicaemia. Infections of the respiratory tract were the causes of other hospitalizations. Two patients also suffered from frequent mild respiratory tract infections; in both patients, decreased immunoglobulin IgA and immunoglobulin IgG2 or immunoglobulin IgG3 levels were recorded. CONCLUSION: Our observations point to an early manifestation of C2 deficiency within the first year of life, with meningitis as the most severe complication. The severity of immunodeficiency may be influenced by concomitant deficiencies of immunoglobulin isotypes. PMID- 12755934 TI - Measured and predicted total body water in children with myelomeningocele. AB - OBJECTIVE: Children with myelomeningocele (MMC) have an altered body composition and an atypical distribution of total body water (TBW). The aim of the present study was to determine the accuracy of current predictive equations, based on bioelectrical impedance analysis (BIA), in determining TBW when compared with measured TBW using deuterium dilution. METHODS: Fourteen children with MMC were measured for whole body BIA and TBW (using deuterium dilution and the Plateau method). Total body water was predicted using equations based on the resistance and characteristic frequency from BIA measurements and heights of subjects. RESULTS: The mean measured TBW was 15.46 +/- 8.28 L and the mean predictions for TBW using equations based on the resistance and characteristic frequency from BIA measurements and heights of subjects were 18.29 +/- 8.41 L, 17.72 +/- 11.42 L and 12.51 +/- 7.59 L, respectively. The best correlation was found using characteristic frequency. The limits of agreement between measured and predicted TBW values using Bland-Altman analysis were large. CONCLUSIONS: The present study suggests that the prediction of TBW in children with MMC can be made accurately using the equation of Cornish et al. based on BIA measurements of characteristic frequency. PMID- 12755935 TI - Pleuro-pulmonary involvement in children with blunt splenic trauma. AB - OBJECTIVE: Evaluation of the importance of pleuro-pulmonary involvement in paediatric patients with blunt splenic trauma. METHOD: A retrospective chart review of 27 patients, aged 2-16 years, treated for blunt splenic injury between 1992 and 1999 was performed. RESULTS: All patients except one were treated conservatively. In 12 patients (44.4%) left-sided pleuro-pulmonary involvement was diagnosed as primary traumatic injury or as a late complication. While Grade I and II splenic injuries were prevalent, pleuro-pulmonary involvement patients had a more severe degree of splenic injury. Chest pain, dyspnoea and diminished respiratory sounds were present on primary examination in patients with chest trauma. Body temperature during the first 5 post-trauma days was significantly higher among pleuro-pulmonary involvement patients. Specific pleuro-pulmonary involvement diagnoses on admission in six children with primary chest trauma were: lung contusion, pleural thickness, or haemo-pneumothorax. Three of them developed delayed pleural effusion. In the other six children with pleuro pulmonary involvement, late complications appeared during 2-5 days post-trauma. CONCLUSIONS: Pleuro-pulmonary involvement was observed in almost half of patients with blunt splenic trauma. Pleuro-pulmonary involvement occurred either early as a result of direct chest trauma or was delayed. High suspicion, careful monitoring of body temperature and repeated chest X-ray studies are recommended for early diagnosis and treatment of delayed pleuro-pulmonary involvement. PMID- 12755936 TI - Effects of stool dilution on the faecal Helicobacter pylori antigen test. AB - OBJECTIVE: To examine the effects of stool dilution on the results of the faecal Helicobacter pylori antigen test. METHODS: Stool samples from nine children with H. pylori infection were diluted with normal saline at dilutions of 1, 5, 10, 50, 100 and 500 times, and optical density (OD) was measured. RESULTS: The faecal H. pylori antigen test yielded positive results for all samples at dilutions of 1:10 and less, although the sample from the case whose original stool showed the lowest OD value changed from positive to negative at a dilution of 1:50. CONCLUSION: Our results suggest that stools at dilutions of less than 1:10 usually do not yield false-negative results in the faecal H. pylori antigen test, even in patients with low faecal antigen levels. PMID- 12755937 TI - Heparin-induced thrombocytopenia in children. AB - OBJECTIVE: To audit the frequency of heparinoid (standard heparin and low molecular weight heparin) use in a tertiary paediatric hospital, and to determine the occurrence of heparin-induced thrombocytopenia (HIT). METHODS: A 1-week cross sectional audit of all heparinoids given to inpatients at a tertiary paediatric hospital was undertaken and a retrospective medical record review of all suspected HIT cases at the tertiary paediatric centre over a 2-year period was carried out. RESULTS: One hundred and sixteen patients received heparinoid medications over a 7-day period. An average of 29 children received heparin daily. The retrospective medical record review identified four patients with suspected HIT over a 2-year period. Two patients developed thrombotic complications, which were fatal in one patient. CONCLUSION: Heparin is used frequently in paediatric tertiary hospitals, yet the occurrence of HIT in children is much lower than that reported in adults. Improved laboratory techniques could facilitate improved screening and diagnosis of this serious adverse drug reaction. PMID- 12755938 TI - Parents' descriptions of development and problems associated with infants with Turner syndrome: a retrospective study. AB - OBJECTIVE: To describe parents' experiences of having an infant diagnosed with Turner syndrome and to determine whether receiving the diagnosis influenced the parents' perceptions of their child's development and/or problems during infancy. In addition, we set out to determine whether the late development of the infant and the perceived problems were related to genotype. METHODS: In this retrospective study, 54 parents (39 mothers and 15 fathers) from different families, each containing a girl with Turner syndrome, were interviewed in order to describe the development, feeding and overall well-being of their daughter during infancy (defined as being before the age of 2 years). RESULTS: Late development was reported to occur in the areas of motor activity (39%), fine motor control (59%), speech (37%) and language (37%). Feeding problems were frequent (74%) and screaming periods occurred in 41%. No differences were found between the responses of the parents whose children were diagnosed before 2 years of age and the responses of those whose children were diagnosed after 2 years of age. No differences were found concerning development and/or problems between the genotypes. CONCLUSIONS: Parents reported delayed development and problems to do with feeding and crying during infancy. These problems had an effect on their everyday life and that of their families, especially the problems relating to feeding. Parents reported that support and advice would have been of significant benefit in coping with the feeding difficulties. Parents were particularly concerned that the personnel at well-baby clinics should be more knowledgeable about the difficulties that can occur in families with an infant with Turner syndrome. PMID- 12755939 TI - Correlates of specific childhood feeding problems. AB - OBJECTIVE: The correlates of specific childhood feeding problems are described to further examine possible predisposing factors for feeding problems. We report our experience with 349 participants evaluated by an interdisciplinary feeding team. METHODS: A review of records was conducted and each participant was identified as having one or more of five functionally defined feeding problems: food refusal, food selectivity by type, food selectivity by texture, oral motor delays, or dysphagia. The prevalence of predisposing factors for these feeding problems was examined. Predisposing factors included developmental disabilities, gastrointestinal problems, cardiopulmonary problems, neurological problems, renal disease and anatomical anomalies. RESULTS: The frequencies of predisposing factors varied by feeding problem. Differences were found in the prevalence of the five feeding problems among children with three different developmental disabilities: autism, Down syndrome and cerebral palsy. Gastro-oesophageal reflux was the most prevalent condition found among all children in the sample and was the factor most often associated with food refusal. Neurological conditions and anatomical anomalies were highly associated with skill deficits, such as oral motor delays and dysphagia. CONCLUSIONS: Specific medical conditions and developmental disabilities are often associated with certain feeding problems. Information concerning predisposing factors of feeding problems can help providers employ appropriate primary, secondary and tertiary prevention measures to decrease the frequency or severity of some feeding problems. PMID- 12755940 TI - Bilious vomiting in a 6-month-old infant. PMID- 12755941 TI - 8: Non-parametric methods for continuous or ordered data. PMID- 12755942 TI - Hyponatraemic hypertensive syndrome in two extremely low birthweight infants. AB - Two cases of hyponatraemic hypertensive syndrome occurring in extremely low birthweight infants are presented. Both infants experienced unilateral renal ischaemia resulting in hyponatraemia and hypertension. A proposed pathophysiological mechanism, namely unilateral renal ischaemia leading to a pressure-natriuresis in the contralateral kidney, is presented. This is associated with an increase in plasma renin and aldosterone, with a paradoxical increase in urinary sodium loss. Immature renal tubular function and relative aldosterone resistance could place the extremely low birthweight infant at increased risk for the condition. The paucity of reports suggests that the condition might be under-recognized. PMID- 12755943 TI - Acute opioid withdrawal on accidental injection of naltrexone. AB - We report two 16-year-old female intravenous drug users who, after making purchases from street suppliers, both presented with symptoms of acute opioid withdrawal. Urine toxicology revealed naltrexone, a long-acting opioid antagonist used in detoxification and maintenance therapy in opioid dependence. While the safety and efficacy of opiate antagonist treatment is being debated, the present case highlights the vulnerability of this young population. The recent availability of non-prescribed opiate antagonists suggests that both health professionals and young people themselves need to be aware of their effects. PMID- 12755944 TI - Transient paralytic ileus following the use of cyclopentolate-phenylephrine eye drops during screening for retinopathy of prematurity. AB - Cyclopentolate-phenylephrine eye drops are commonly used for mydriasis during routine screening for retinopathy of prematurity in preterm infants. Although systemic absorption is minimal, it can result in side effects. We report two cases of transient paralytic ileus associated with transient oxygen desaturation and hypertension following the use of cyclopentolate-phenylephrine eye drops. PMID- 12755953 TI - Multiple endocrine neoplasia. PMID- 12755954 TI - Multiple endocrine neoplasia type 1: duodenopancreatic tumours. AB - The pancreaticoduodenal disease in Multiple endocrine neoplasia type 1 (MEN1) is the most frequent cause of death due to the syndrome, and the most controversial with regard to its management. This article discusses the current data and recommendations with respect to disease screening, functional tumour diagnosis, natural history, preoperative imaging, operative strategy and follow-up. PMID- 12755955 TI - The treatment of sporadic versus MEN1-related pituitary adenomas. AB - The treatment of pituitary tumours strongly depends on their clinical presentation. In general, the treatment aims are reducing tumour volume and/or decreasing hormone hypersecretion. It relies on single or a combination of three different methods: surgery, medication and radiotherapy. The rationale for deciding the treatment are many but include the aggressiveness of the tumour. The aetiologies of sporadic pituitary adenomas are not fully understood. However, several causes have been identified resulting in specific familial phenotypes like multiple endocrine neoplasia type I (MEN1). MEN1 is related to mutations in the MEN1 gene, a tumour suppressor gene localized on chromosome 11q13 and which encodes menin, a 610 amino acid protein. During the last years, an evidence progressively emerged that MEN1-related adenomas were more aggressive and less responsive to therapy than their sporadic counterparts. In this article, we review the differences between sporadic and MEN1-related adenomas and suggest specific ways of treatment and follow-up for MEN1-related tumours. PMID- 12755956 TI - Functional studies of the MEN1 gene. AB - Multiple endocrine neoplasia type 1 is an autosomal dominant cancer syndrome affecting primarily parathyroid, enteropancreatic endocrine and pituitary tissues. The inactivating germline and somatic mutations spread throughout the gene and the accompanying loss of the second allele in tumours show that the MEN1 gene is a tumour suppressor. The MEN1-encoded protein, menin, is a novel nuclear protein. Menin binds and alters JunD-, NF-kappaB-, Smad3-mediated transcriptional activation. The mouse Men1 knockout model mimicks the human MEN1 condition contributing to the understanding of tumorigenesis in MEN1. PMID- 12755957 TI - Surgical treatment of medullary thyroid carcinoma. AB - Medullary thyroid carcinoma (MTC) is a malignancy of the parafollicular C cells of the thyroid gland. It occurs sporadically or as part of the multiple endocrine neoplasia type 2 (MEN 2) syndromes. Patients who have inherited a mutation in the RET proto-oncogene should have thyroidectomy early in life to prevent formation and spread of this cancer. Most patients with sporadic disease present with a palpable neck mass. The diagnosis is made by fine needle aspiration biopsy and by measuring calcitonin levels in the blood. Primary treatment consists of surgical resection including a total thyroidectomy, central neck nodal dissection and functional lateral neck nodal dissections. Most patients with a palpable primary tumour have nodal disease present at the time of operation, and nodal involvement is often bilateral. Adequate resection of the primary tumour and cervical lymph nodes is important to optimize outcome and minimize the risk of recurrent disease. Proper handling of the parathyroid glands prevents hypoparathyroidism. Following primary surgical resection, more than half of the patients will have recurrent disease with persistent elevation of calcitonin levels. Currently, there is no adequate systemic therapy for treating recurrent disease. Surgical reoperation or conservative observation are the best available options. Diagnostic laparoscopy for liver evaluation is the most sensitive diagnostic test to detect the presence of distant metastases. PMID- 12755958 TI - Cell signalling and gene expression mediated by RET tyrosine kinase. AB - Germline mutations of the RET proto-oncogene cause multiple endocrine neoplasia (MEN) 2A or 2B by different mechanisms. As is the case for other receptor tyrosine kinases, mutant RET recruits a variety of signalling molecules via phosphorylated tyrosine residues present in the kinase domain and carboxy terminal tail. As we previously reported, the signaling via phosphorylated tyrosine 1062 plays a crucial role in the transforming activities of both RET MEN2A and RET-MEN2B mutant protein. Interestingly, this single tyrosine residue represents a binding site for several signalling molecules including SHC, Enigma, SNT/FRS2, DOK and IRS1 and is responsible for activation of the RAS/ERK, PI3 K/AKT, JNK, p38MAPK and ERK5 signalling pathways. Amongst these, the PI3-K/AKT and JNK pathways appeared to be more strongly activated in the cells expressing RET-MEN2B than in the cells expressing RET-MEN2A, suggesting the possibility that these pathways may be involved in the disease phenotype. In addition, RET is alternatively spliced to produce three isoforms and the splicing site is present just downstream of tyrosine 1062. These isoforms play different roles for the tumour development associated with MEN 2 or the development of the kidney and the enteric nervous system. Moreover, using differential display analysis, we identified several genes whose expression is highly induced by RET-MEN2B mutant proteins. The differential gene expression by RET-MEN2A and RET-MEN2B may also be important for the development of their phenotypes. PMID- 12755959 TI - Hyperparathyroidism-jaw tumour syndrome. AB - Amongst hyperparathyroidism-related syndromes, hyperparathyroidism-jaw tumour syndrome is one of the least common and relatively unknown but its clinical and genetic aspects are not less interesting or important. With the recent identification of its genes, we can now better characterize the disease, both clinically and genetically, which will certainly impact the field of endocrinology and oncology. In this article, we review the clinico-pathological features and genetic basis of this syndrome with the hope that it will create awareness and interest in this disease amongst clinicians and basic scientists. PMID- 12755960 TI - Adverse renal effects of anti-inflammatory agents: evaluation of selective and nonselective cyclooxygenase inhibitors. AB - Conventional nonsteroidal anti-inflammatory drugs (NSAIDs), i.e. nonselective cyclooxygenase COX inhibitors have well-documented nephrotoxicity. Adverse renal effects occur because of inhibition of the synthesis of cyclooxygenase-derived prostaglandins which act to modulate pathologic processes that would normally impair various renal functions. The introduction of the selective COX-2 inhibitors raised hope that this class of drugs would reduce injury in both the gastrointestinal tract and the kidneys. Animal and human data, however, suggest that COX-2 synthesized prostaglandins are important in the modulation of renal physiology during adverse conditions. Hence, it appears that these drugs are equal in causing nephrotoxicity as the nonselective COX inhibitors. PMID- 12755961 TI - Association of boiled and filtered coffee with incidence of first nonfatal myocardial infarction: the SHEEP and the VHEEP study. AB - OBJECTIVES: To evaluate the influence of consumption of filtered and boiled coffee, on the incidence of first nonfatal myocardial infarction. DESIGN: Population-based case-control study. SETTING AND SUBJECTS: The study base consisted of the population 45-65/70 years-old in two Swedish counties, Stockholm and Vasternorrland, 1992/93-94. In all, 1943 cases of first nonfatal myocardial infarction were identified. For each case one control was selected from the study base concurrently with disease incidence by matching the sex, age and place of residence of the case. Information about coffee consumption and other factors was obtained by mailed questionnaire and a medical examination. The participation rate was 85% amongst cases and 74% amongst controls. RESULTS: Men with a reported consumption of 7-9 dL filtered coffee per day showed an increased incidence of first myocardial infarction compared with consumers of 3 dL day-1 or less (RR: 1.32; 95% CI: 1.03-1.70). A consumption of at least 10 dL day-1 was associated with an RR of 1.93 (95% CI: 1.42-2.63) for filtered and 2.20 (95% CI: 1.17-4.15) for boiled coffee. Amongst women, no clear association was seen between consumption of filtered coffee and myocardial infarction but consumption of boiled coffee tended to be related to an increased incidence. Comparing subjects drinking boiled coffee with those drinking filtered coffee and adjusting for the amount consumed gave an increased incidence for boiled coffee amongst both men (RR: 1.41; 95% CI: 1.07-1.80) and women (RR: 1.63; 95% CI: 1.04-2.56). CONCLUSIONS: Consumption of boiled coffee appears to increase the incidence of first nonfatal myocardial infarction. This increased incidence is consistent with randomized trials showing an adverse impact of boiled coffee on blood lipids. PMID- 12755962 TI - Evolving patterns of tobacco use in northern Sweden. AB - BACKGROUND AND OBJECTIVES: Cross-sectional data from northern Sweden suggest that the increased use of Swedish moist snuff (snus) may have contributed to a decline in the prevalence of smoking, especially amongst men. This study describes the evolving patterns of tobacco use in this population over the period 1986-1999. DESIGN: This is a prospective follow-up study of 1651 men and 1756 women, aged 25 64 years, who were enrolled in the northern Sweden MONICA project (entry in 1986, 1990, 1994) and who were followed-up in 1999. Information on tobacco use at entry and at follow-up was used to describe the stability of tobacco use over a period of 5-13 years ending in 1999. RESULTS: Snus was the most stable form of tobacco use amongst men (75%); only 2% of users switched to cigarettes and 20% quit tobacco altogether. Smoking was less stable (54%); 27% of smokers were tobacco free and 12% used snus at follow-up. Combined use (smoking and snus) was the least stable (39%), as 43% switched to snus and 6% switched to cigarettes. Former users of both products were much less stable than former users of either cigarettes or snus. The stability of smoking amongst women was 69%, which was higher than that amongst men (P < 0.05). CONCLUSIONS: The use of snus played a major role in the decline of smoking rates amongst men in northern Sweden. The evolution from smoking to snus use occurred in the absence of a specific public health policy encouraging such a transition and probably resulted from historical and societal influences. PMID- 12755963 TI - The Arctic Oscillation and incidence of acute myocardial infarction. AB - OBJECTIVES: To describe the relation between the Arctic Oscillation (AO) index and the incidence and mortality in acute myocardial infarction (AMI) in the northern, partly subarctic area of Sweden. DESIGN: Comparison of a time series of daily variations in the AO index and register data on the daily number of fatal and nonfatal AMIs. SETTING: The northernmost two Swedish counties, Norrbotten and Vasterbotten. SUBJECTS: All inhabitants in the Norrbotten and Vasterbotten counties were followed for the occurrence of an AMI between 1985 and 1999 within the framework of the WHO MONICA (multinational MONItoring of trends and determinants of CArdiovascular disease) Project. MAIN OUTCOME MEASURE: Fatal and nonfatal AMIs. RESULTS: There was a consistent positive relation between increasing AO index and an increase in AMI incidence and mortality. The maximum impact on AMI incidence of the AO came after a lag phase of 3 days. A one unit increase in AO index was associated with an increase in: the daily number of AMIs (+3.8%), the case fatality in AMI within 28 days (+5.1%), the number of nonfatal AMIs (+3.4%), and the number of sudden cardiac deaths (+8.3%). CONCLUSIONS: An AO index increase, bringing warmer weather over Scandinavia, was associated with an increase in the incidence and mortality in AMI in northern Sweden. PMID- 12755965 TI - Modigliani's "fillette en bleu": a case of juvenile dermatomyositis? PMID- 12755966 TI - Trichotillomania. PMID- 12755967 TI - Azathioprine: current status and future considerations. PMID- 12755968 TI - Lichen planopilaris: report of 30 cases and review of the literature. AB - BACKGROUND: Lichen planopilaris (LPP) affects primarily the scalp, resulting in scaling, atrophy, and alopecia with scarring. The purpose of our study was to obtain original data on LPP and to evaluate the efficacy of topical therapy in comparison with systemic therapies. METHODS: We examined 30 patients affected by LPP between 1996 and 2001, performing clinical, laboratory, histopathologic and direct immunofluorescence examinations. Twenty-one of the patients (70%) were women and nine (30%) were men. The average age at presentation was 51.5 years. The average duration of the disease was 13 months at the time of the diagnosis. All patients received topical steroids for a total of 12 weeks. RESULTS: Resolution of the inflammatory process and blocking of the cicatricial progression were observed in 66% of cases, a mild reduction of fibrosis and cicatrization in 20% of patients, and no response in 13%. CONCLUSIONS: We concluded that topical therapy may be a valid alternative to systemic therapies, especially in patients with lesions in the early phase. PMID- 12755969 TI - Evaluation of clinical findings according to sex in 2313 Turkish patients with Behcet's disease. AB - BACKGROUND: Behcet's disease is a multisystem disease featuring mucocutaneous, ocular, articular, vascular, intestinal, urogenital, and neurologic involvement and occurs with a high prevalence in the Mediterranean including Turkey. Higher incidence of severe clinical course and systemic involvement is observed in male patients. OBJECTIVE: To determine the influence of sex on the clinical course of Behcet' s disease. METHODS: We retrospectively evaluated the clinical findings of 2313 Behcet patients followed up at the multidisciplinary Behcet's Disease Center at Ankara University. RESULTS: The male/female patient ratio was 1.03. Oral aphthae was seen in all patients. In male Behcet patients, the prevalence of mucocutaneous lesions and systemic manifestations was as follows: 85.6% genital aphthae, 45.5% erythema nodosum, 59.5% papulopustular lesions, 17.5% thrombophlebitis, 38.1% ocular involvement, 11.3% articular involvement, 11.7% vascular involvement, 3.3% neurologic involvement, 1.4% gastrointestinal involvement, and 1.8% pulmonary involvement. In female Behcet patients, the prevalence of manifestations were as follows: 91% genital aphthae, 49.8% erythema nodosum, 48.3% papulopustular lesions, 3.5% thrombophlebitis, 19.8% ocular involvement, 11.8% articular involvement, 2.1% vascular involvement, 1.3% neurologic involvement, 1.4% gastrointestinal involvement, and 0.03% pulmonary involvement. CONCLUSIONS: Only genital aphthae and erythema nodosum were more frequent in females. On the other hand papulopustular eruptions, thrombophlebitis, ocular, neurologic, pulmonary and vascular involvement were more frequent in males. While female patients had the best prognosis, male patients had a worse overall prognosis than females. PMID- 12755970 TI - Sunscreen use and skin protection behavior on the Belgian beach. AB - BACKGROUND: Public health campaigns encourage the public to protect themselves against skin cancer by using sunscreens and taking other protective measures. However, it is difficult to estimate the level of awareness among the general public. METHODS: This study explores the prevalence and predictors of solar protection behavior in a community sample of beachgoers. During the months of May, June, July and August 2001 a total of 360 participants (145 men and 215 women) were randomly selected from several Belgian beaches in and around the city of Ostend, Belgium. The solar protection behavior of each participant was assessed by direct observation and interview. RESULTS: The risk awareness percentages were 70.6% for skin cancer, 60.8% for sunburn and 25.0% for skin aging. These percentages were considerably higher in the female participant group than in the male group. Sunscreen cream was the most popular preventive behavior, especially in the female population, whereas the alternative protective measures (limited exposure during peak sun hours, the use of shade and the use of protective clothing and hats) were more popular in the male group. CONCLUSIONS: While solar protection has become part of routine beach behavior, there is room for improvement by more frequent application of sunscreen cream, the use of a higher sun protection factor (SPF) (15+), timed sunbathing, more use of clothing and hats and more seeking of shade. The results of this study can assist in evaluating the effectiveness of current sun-protection campaigns and health education programs. PMID- 12755971 TI - Multiple nodular plexiform neurofibromas in a neurofibromatosis 1 family: a familial tendency? A case report and review of the literature. PMID- 12755972 TI - Ultraviolet radiation properties as applied to photoclimatherapy at the Dead Sea. AB - BACKGROUND: The Dead Sea basin, the lowest terrestrial point on earth, is recognized as a natural treatment center for patients with various cutaneous and rheumatic diseases. Psoriasis is the major skin disease treated at the Dead Sea with excellent improvement to complete clearance exceeding 85% after 4 weeks of treatment. These results were postulated to be associated with a unique spectrum of ultraviolet radiation present in the Dead Sea area. METHODS: The UVB and UVA radiation at two sites is measured continuously by identical sets of broad-band Solar Light Co. Inc. meters (Philadelphia, PA). The spectral selectivity within the UVB and UVA spectrum was determined using a narrow-band spectroradiometer, UV Optronics 742 (Orlando, FL). The optimum exposure time intervals for photoclimatherapy, defined as the minimum ratio of erythema to therapeutic radiation intensities, were also determined using a Solar Light Co. Inc. Microtops II, Ozone Monitor-Sunphotometer. RESULTS: The ultraviolet radiation at the Dead Sea is attenuated relative to Beer Sheva as a result of the increased optical path length and consequent enhanced scattering. The UVB radiation is attenuated to a greater extent than UVA and the shorter erythema UVB spectral range decreased significantly compared with the longer therapeutic UVB wavelengths. CONCLUSIONS: It was demonstrated that the relative attenuation within the UVB spectral range is greatest for the shorter erythema rays and less for the longer therapeutic UVB wavelengths, thus producing a greater proportion of the longer therapeutic UVB wavelengths in the ultraviolet spectrum. These measurements can be utilized to minimize the exposure to solar radiation by correlating the cumulative UVB radiation dose to treatment efficacy and by formulating a patient sun exposure treatment protocol for Dead Sea photoclimatherapy. PMID- 12755973 TI - Increased hydrogen peroxide generation by neutrophils from patients with acne inflammation. AB - BACKGROUND: Reactive oxygen species generated by neutrophils are closely correlated with the pathogenesis of a variety of inflammatory skin diseases. The aim of this study was to investigate the possible role of reactive oxygen species generated by neutrophils in the mediation of acne inflammation. METHODS: Bacterial phagocytotic stimuli, mediated by opsonin activity, were applied to whole blood, and neutrophil hydrogen peroxide production was measured. RESULTS: Patients with acne inflammation showed a significantly increased level of hydrogen peroxide produced by neutrophils compared to patients with acne comedones and healthy controls. There were no marked differences in the level of hydrogen peroxide produced by neutrophils between patients with acne comedones and healthy controls. In addition, patients with acne inflammation treated by oral administration of minocycline hydrochloride, a drug that inhibits hydrogen peroxide generation by neutrophils, showed a significant decrease in the ability of neutrophils to produce hydrogen peroxide in accordance with a decrease in the inflammatory activity of acne lesions. CONCLUSIONS: The present study seems to suggest that acne inflammation is mediated in part by hydrogen peroxide generation by neutrophils. PMID- 12755974 TI - Bromoderma. AB - A 3-year-old Japanese girl with severe epilepsy had been treated with potassium bromide since August 1999. The dose of potassium bromide was increased from 0.5 g/day to 0.8 g/day in May 2000 because of poor control of epilepsy. She also presented high fever, caused by bacterial pneumonia, in the same period. On June 11, a reddish eruption suddenly appeared on her back. Physical examination revealed grain-size, dark-red, erythematous papules and pustules on the back and face (Fig. 1). Some of the lesions on the back were ovoid to circular with small pustules and necrotic centers. Although some papules seemed to have dell in their centers, showing the appearance of herpes virus infection, Tzanck test was negative. A biopsy specimen obtained from one of the papules revealed a massive infiltration of eosinophils and neutrophils, forming an abscess in the epidermis and dermis (Fig. 2). The serum bromide level, which was 43.7 mEq/L (normal, 0-5 mEq/L) on May 25, increased to 114 mEq/L on June 14. The eruption disappeared within 10 days after the withdrawal of potassium bromide and treatment with topical sulfadiazine silver cream. The serum bromide decreased to 56.8 mEq/L on July 6. PMID- 12755975 TI - Treatment of prurigo nodularis with thalidomide: a case report and review of the literature. PMID- 12755976 TI - Linear ectodermal cutaneous hamartoma. PMID- 12755977 TI - Erythema induratum of bazin: role of polymerase chain reaction in diagnosis. PMID- 12755979 TI - A case of subcutaneous nodular fat necrosis with lipase-secreting acinar cell carcinoma. PMID- 12755980 TI - Churg-Strauss syndrome manifested by urticarial plaques. PMID- 12755978 TI - Axillary intertriginous granular parakeratosis responsive to topical calcipotriene and ammonium lactate. PMID- 12755981 TI - Acute onset of neutrophilic dermatosis in patients after therapy with a COX-2 specific inhibitor. PMID- 12755982 TI - Recurrent aphthous stomatitis unresponsive to topical corticosteroids: a study of the comparative therapeutic effects of systemic prednisone and systemic sulodexide. AB - BACKGROUND: Recurrent aphthous stomatitis (RAS) is a common oral condition, the etiology of which remains largely unclear. Numerous therapeutic protocols have been tried. Apart from immunomodulators, no therapy is unequivocally effective, and many systemic therapies have potential adverse effects. OBJECTIVE: To compare, in patients with frequent RAS unresponsive to conventional therapies, the therapeutic effectiveness of systemic prednisone with that of systemic sulodexide, a low-molecular-weight heparin with immunosuppressive activity but few adverse effects. METHODS: The study involved a group of 30 patients suffering from frequent minor RAS over >or= 4 months unresponsive to topical corticosteroids. Patients were randomly assigned to one of three study groups: blind therapy with systemic sulodexide or systemic prednisone and control (no treatment). The outcomes were assessed blind on the basis of the days to recovery from pain and days to recovery from ulceration (epithelialization) during the first month of therapy; the number of aphthae appearing during the second month of therapy; and the number of aphthae appearing in the 2 months after the end of the 2-month treatment cycle. RESULTS AND CONCLUSIONS: The effectiveness of systemic sulodexide was almost comparable with that of systemic prednisone in patients with frequent RAS, without significant adverse effects. PMID- 12755983 TI - Achievements of dermatological research in Denmark and Israel: a comparative 10 year study. AB - Wide differences of achievement in dermatological research between Denmark and Israel have been reported, although the two countries are comparable in terms of academic dermatological structure. The aims of the present study were to document these differences by means of bibliometric analysis, and to attempt to elucidate the causes of these differences. Employing MEDLINE searches for the 10-year period 1988-97, quantitative and qualitative comparisons of the dermatological publications from these two countries were conducted. We found the achievements of Danish dermatological research to be superior to those of Israel, and suggest that the large proportion of case reports and reviews is one cause of the relatively low ranking of Israeli dermatological research efforts. PMID- 12755984 TI - Ulcers caused by bullous morphea treated with tissue-engineered skin. AB - Bullous morphea is an uncommon form of localized scleroderma. The exact pathogenesis is unknown and treatment of the accompanying ulcers is problematic. We report a patient with bullous morphea with long-standing ulcers whom we successfully treated with the tissue-engineered skin Apligraf (Organogenesis Inc., Canton, MA). The patient experienced rapid improvement in granulation tissue and the ulcers healed 4 months after a single application. The rationale for the use of Apligraf is based on experience with patients with venous ulcers who have surrounding peri-ulcer fibrosis. This condition, termed lipodermatosclerosis, has been reported as a poor prognostic factor for healing, yet many ulcers associated with lipodermatosclerosis may respond to treatment with tissue-engineered skin. Taken in concert, these results suggest a role for tissue- engineered skin in the treatment of chronic wounds with surrounding fibrosis. PMID- 12755985 TI - The use of the freer dissector for the removal of trichilemmal cysts. AB - BACKGROUND: Trichilemmal cysts are keratin-containing cysts, usually situated on the scalp. They often show an autosomal dominant inheritance pattern. Excision is the treatment of choice. We describe a practical surgical technique that eases the removal of these cysts. METHODS: A 24-year-old woman presented with a 5-year history of cystic lesions on her scalp, clinically diagnosed as trichilemmal cysts. She was treated with the new technique. After making a small incision under local anesthesia, a freer dissector (Aesculap(R) OL 165 R) was used as a blunt dissector. The freer dissector was inserted through the incision. A blunt dissection was made to dissect the cyst free from the surrounding dermis and, by using the dissector as a lever, gentle pressure was applied to the opposite side to ease the cyst from the dermis. Four cysts were removed with this technique. RESULTS: The treatment was well tolerated by the patient. No complications developed during or after the procedure. CONCLUSIONS: This modified technique is simple and practical. The slight curve at each end of the freer dissector makes it easy to grasp the cyst and, functioning like forceps, the cyst can be eased from the dermis. We believe that the use of the freer dissector in the surgical excision of trichilemmal cysts aids and speeds up the procedure. PMID- 12755986 TI - Treatment of chromomycosis by cryosurgery with liquid nitrogen: 15 years' experience. AB - BACKGROUND: Chromomycosis is a chronic fungal infection of the skin and subcutaneous tissue caused by various dematiaceous fungi, the most common of which is Fonsecaea pedrosoi. The disease is difficult to treat. METHODS: Twenty two chromomycosis patients treated by cryosurgery with liquid nitrogen as the sole or associated therapy were studied. Diagnosis was confirmed by direct examination of KOH cleared specimens, culture, and histology, according to a method previously described by Pimentel et al. (Pimentel ERA, Castro LGM, Cuce LC, Sampaio SAP. Treatment of chromomycosis by cryosurgery with liquid nitrogen. J Dermatol Surg Oncol 1989; 15: 72-77). Small lesions were frozen in a single session, whilst larger lesions were frozen in small parts. All patients received double freeze-thaw cycles. An author-created severity index, based on the extent of the diseased area, number of lesions, presence of complications (lymphedema, ulceration, and secondary infection) and unresponsiveness to previous treatments, was used to determine disease severity. RESULTS: Fonsecaea pedrosoi was isolated from all but one patient. Caucasian (50.0%) males (4.5 : 1.0) were most commonly affected. The average number of cryosurgery sessions per patient was 6.7, varying from one to 22 sessions. The duration of treatment lasted for up to 126 months. Nine patients (40.9%) were considered to be cured (clinically disease-free period of at least 3 years), eight (36.4%) were under observation (clinically disease free but less than 3 years of follow-up), two (9.1%) were under treatment (still with active lesions), and three (13.6%) were classified as unsuccessful. The average cure period was 7.5 years, varying from 3 to 16 years. Five (55.6%) of the nine cured patients had been cured for periods exceeding 9 years. Six (66.7%) of the nine cured patients were classified as having mild, two (22.2%) severe, and one (11.1%) moderate disease. CONCLUSIONS: Cryosurgery with liquid nitrogen is an option in the treatment of chromomycosis. PMID- 12755987 TI - A new grading system for oral pemphigus. PMID- 12755988 TI - A case of hypertrophic lichen ruber planus of the leg complicated by a squamous cell carcinoma. PMID- 12755989 TI - Prurigo pigmentosa is distinctive histopathologically. PMID- 12755990 TI - Pathophysiology of erectile dysfunction. PMID- 12755991 TI - Erectile dysfunction after surgical treatment. AB - Erectile dysfunction is a recognized complication of prostate and bladder radical surgery, although there is significant variation in the reported risk, much of this variability is related to the retrospective nature of most previous studies. Undoubtedly, the quality of life of bladder and prostate cancer patients would be much improved if both normal micturition and potency are preserved, which is the subject of this article. Quality of life studies can delineate sexual function after radical prostatectomy, including the use of sexual aids. Penile erection is a neurovascular event modulated by neurotransmitters and hormonal status. The penis is innervated by autonomic and somatic nerves. Both surgery and radiation therapy appear to affect such a mechanism. Radiation is thought to produce Erectile Dysfunction (ED) by accelerating microvascular angiopathy causing cavernosal fibrosis or stenosis of the pelvic arteries and by accelerating existing arteriosclerosis, leading to vascular impotence. Years may elapse before clinically significant ED occurs. Criteria that influence recovery of erections after surgery include younger patient age, stronger erections before operation, preservation of the neurovascular bundles, and attention to fine details in the surgical technique. Recovery of erections occurs in 68% of preoperatively potent men treated with bilateral nerve-sparing surgery and in 47% of those treated with unilateral nerve-sparing surgery. PMID- 12755992 TI - Suppression of testosterone stimulates recovery of spermatogenesis after cancer treatment. AB - It is important to develop methods to prevent or reverse the infertility caused by chemotherapy or radiation therapy for cancer in men. Radiation and some chemotherapeutic agents kill spermatogonial stem cells, but we have shown that these cells survive in rats, although they are unable to differentiate. There is evidence that this phenomenon also occurs in men. The block to spermatogonial differentiation in rats is caused by some unknown change, either in the spermatogonia or the somatic elements of the testis, such that testosterone inhibits spermatogonial differentiation. In the rat, the spermatogenesis and fertility lost following treatment with radiation or some chemotherapeutic agents can be restored by suppressing testosterone with gonadotropin releasing hormone (GnRH) agonists or antagonists, either before or after the cytotoxic insult. The applicability of this procedure to humans is still unknown. Some anticancer regimens may kill all the stem cells, in which case the only option would be spermatogonial transplantation. However, in some cases stem cells survive and there is one report of stimulation of recovery of spermatogenesis with hormonal treatment. Clinical trials should focus on treating patients with hormones during or soon after anticancer treatment. The hormone regimen should involve suppression of testosterone production with minimum androgen supplementation used to improve the diminished libido. PMID- 12755993 TI - TGF-betas: their role in testicular function and Sertoli cell tight junction dynamics. AB - Transforming growth factor-betas (TGF-betas) are known to regulate multiple physiological functions in the testis, which include spermatogenesis, Leydig cell steroidogenesis, extracellular matrix synthesis and testis development. More recent studies have shown that TGF-beta3 also regulates Sertoli cell tight junction (TJ) dynamics in vitro via the p38 mitogen-activated protein (MAP) kinase pathway, suggesting that this cytokine plays a crucial role in regulating the opening and closing of the blood-testis barrier (BTB). This in turn regulates the passage of pre-leptotene and leptotene spermatocytes across the BTB at stages VIII-XI of the seminiferous epithelial cycle. This review summarizes recent advances of studies on TGF-betas in the testis, highlighting their regulatory role in TJ dynamics. PMID- 12755994 TI - Male age is not an independent factor to affect the outcome of assisted reproductive techniques. AB - Controversy exists whether advanced male age is associated with poor sperm quality and subsequent failure in the assisted reproductive techniques (ART). The purpose of this study was to evaluate the effect of male age on sperm quality and the outcome of ART as well as the association of male age with other relevant factors, particularly with the female age. A retrospective study was performed in order to evaluate the effect of male age on the sperm parameters in 880 routine seminal analyses. Additionally, sperm parameters were also compared among different age groups in 919 cases with male factor infertility who had been included in an ART programme. The laboratory and clinical results of ART (fertilization rate, number and quality of embryos transferred, as well as pregnancy rates) were compared according to different age groups. The results were also evaluated by one-way correlation and also step-wise logistic regression analysis to identify the interactions and correlations between different parameters. There were no statistically significant differences between male age groups in terms of sperm concentration, motility and morphology either in routine seminal analyses or in ART groups. In the ART group, a statistically significant linear correlation was present between male and female ages. Male age was increasing in parallel to female age. Female age was also correlated significantly with ART results. In one-way correlation analysis, male age was found to be correlated with the pregnancy rate, but not with fertilization rate and the quality of the transferred embryos. However, regression analysis revealed that correlation between male age and pregnancy results was simply dependent on the effect of the female age. Seminal parameters did not reveal a significant change with the increasing male age. The effect of male age on ART results in cases with male factor infertility is not a direct effect but a reflection of the negative impact of the parallel increase in the female age. PMID- 12755996 TI - Effects of hepatocyte growth factor on urokinase-type plasminogen activator (uPA) and uPA receptor in DU145 prostate cancer cells. AB - Urokinase-type plasminogen activator (uPA) and the uPA receptor (uPAR) are involved in a proteolytic cascade resulting of extracellular matrix degradation. Upstream, uPA and uPAR are regulated by various factors including hepatocyte growth factor (HGF), which stimulates the uPA/uPAR proteolytic system and increases invasion of cancers. We recently demonstrated that HGF induces invasion of DU145 prostate cancer cells into collagen gel matrix. We therefore examined effects of HGF on uPA and uPAR expression in DU145 cells. Effects of HGF on uPA expression in culture medium were determined by Western blotting and fibrin zymography, effects on uPAR expression in cell-associated protein were examined by Western blotting. HGF increased uPA and uPAR production in a dose-dependent manner up to 10 ng/mL, while effects of 20 ng/mL were approximately equal to those of 10 ng/mL. HGF stimulated uPA production beyond that in control cultures from 8 h until 48 h after HGF addition. HGF stimulated a uPA/uPAR proteolytic network in DU145 cells, which may be important for acquisition invasive potential by prostate cancer. PMID- 12755995 TI - Microtubule-associated epithelial protein E-MAP-115 is localized in the spermatid manchette. AB - A microtubule-associated protein E-MAP-115 has been originally isolated and characterized from HeLa cells. Because of its predominant expression in cultured cells of epithelial origin, it has been suggested to be involved in the regulation of cell polarization. The present immunocytochemical, Northern blot and in situ hybridization analysis of E-MAP-115 in the mouse and rat seminiferous epithelium indicates its distinct association with the spermatid manchette, a unique microtubular structure which appears in the cytoplasm of spermatids at step 8 when nuclear polarization and elongation starts. At steps 15-16 when manchette has been disassembled, immunoreactivity for E-MAP-115 disappeared. At immunoelectron microscopical level, E-MAP-15 was associated with the microtubules of the manchette. In the Western and Northern blot analysis, a distinct stage dependent expression of a single E-MAP-115 polypeptide and two mRNA species (3.4 and 2.4 kb) could be identified. MTEST 60, a spermatid-specific transcript, showed a 100% homology over region of 68-193 bp of E-MAP-115 sequence. The reported specific localization of E-MAP-115 to the spermatid manchette strongly supports its role as a regulator of cell polarization. This, in turn, supports the hypotheses concerning the dynamic function of the manchette during spermiogenesis. PMID- 12755997 TI - A stereological study of the effects of experimental inguinal cryptorchidism and subsequent orchiopexy on spermatogenesis in adult monkeys. AB - Our previous study demonstrated that experimental intra-abdominal cryptorchidism in adult rabbits for 13 weeks resulted in severe spermatogenic arrest: type A spermatogonia was the only germ cell type seen in the seminiferous epithelium and its number per testis was reduced by 84%. Seven weeks following orchiopexy, the type A spermatogonial number returned to the near-normal range in most animals and spermatogenesis partially recovered (Reproduction 2002, 124, 95-105). This study aimed to determine whether inguinal cryptorchidism would produce less severe damage to spermatogenesis and whether subsequent orchiopexy would better restore spermatogenesis. Five normal adult male rhesus monkeys (Macaca mulatta) underwent bilateral artificial inguinal cryptorchidism. Half a year later, one testis together with the ipsilateral epididymis were removed from each animal and then unilateral orchiopexy was performed on the contralateral side, with the remaining testis and epididymis being removed another half a year later. A contemporary unbiased and efficient stereological tool, the optical disector, was used to estimate numbers of all types of spermatogenic cells in the testis and spermatozoa in the epididymis. Spermatogenic arrest was induced by cryptorchidism at the stage of spermatogonia (n = 1), spermatocytes (n = 2) or early spermatids (n = 1), with the type A spermatogonial numbers per testis being reduced to 14.8 57.2% of the control average; in one of the five cryptorchid animals, however, spermatogenesis remained normal. Subsequent orchiopexy, which was successfully performed on two animals with cryptorchidism-induced spermatogenic arrest, brought on a full or partial recovery of spermatogenesis. In conclusion, inguinal cryptorchidism induces less severe (in comparison with an intra-abdominal one) and variable damage to spermatogenesis, which is restored, at least in part, by subsequent orchiopexy. PMID- 12755998 TI - The polymorphic androgen receptor gene CAG repeat, pituitary-testicular function and andropausal symptoms in ageing men. AB - The activity of androgen receptor (AR) is modulated by a polymorphic CAG trinucleotide repeat in the AR gene. In the present study, we investigated hormonal changes among ageing men, and whether the number of AR CAG triplets is related to the appearance of these changes, as well as symptoms and diseases associated with ageing. A total of 213 41-70-year-old men donated blood for hormone analyses (LH, testosterone, oestradiol and SHBG) and answered questions concerning diseases and symptoms associated with ageing and/or androgen deficiency. Of these men, 172 donated blood for the measurement of the CAG repeat length of AR. The CAG repeat region of the AR gene was amplified by polymerase chain reaction (PCR) and the products were sized on polyacrylamide gels. The repeat number was analysed as a dichotomized variable divided according to cut off limits of the lowest (< or =20 repeats) and the highest quartile (> or =23 repeats), and as a continuous variable. The proportion of men with serum LH in the uppermost quartile (>6.0 IU/L) with normal serum testosterone (>9.8 nmol/L, above the lowest 10%) increased significantly with age (p = 0.01). There were fewer men with this hormonal condition among those with CAG repeat number in the uppermost quartile (> or =23 repeats) (p = 0.03). These men also reported less decreased potency (p < 0.05). The repeat number was positively correlated with depression, as expressed by the wish to be dead (r = 0.45; p < 0.0001), depressed mood (r = 0.23; p = 0.003), anxiety (r = 0.15; p < 0.05), deterioration of general well-being (r = 0.22; p = 0.004), as well as decreased beard growth (r = 0.49; p < 0.0001). A hormonal condition where serum testosterone is normal but LH increased is a frequent finding in male ageing. Only certain types of age-related changes in ageing men were associated with the length of the AR gene CAG repeat, suggesting that this parameter may play a role in setting different thresholds for the array of androgen actions in the male. PMID- 12756000 TI - Comparing zonal and CFD model predictions of isothermal indoor airflows to experimental data. AB - It is inappropriate to use the assumption of instantaneously well-mixed zones to model airflows and pollutant transport in large indoor spaces. We investigate two approaches for describing the details of airflows in large indoor spaces, for accuracy and suitability for integration with multi-zone infiltration models. One approach, called the zonal method, was developed over the last 15 years to provide an improvement over the well-mixed assumption. The second approach is the use of a computational fluid dynamics simulation using a coarse grid model of the large indoor space. We compare velocity predictions from different formulations of zonal methods and coarse-grid k-epsilon computational fluid dynamics (CFD) models, to measurements, in a 2D mechanically ventilated isothermal room. Our results suggest that, when airflow details are required, coarse-grid CFD is a better-suited method to predict airflows in large indoor spaces coupled with complex multi-zone buildings, than are the zonal methods. Based on the comparison of pressure predictions from different models, we offer guidance regarding the coupling of a model of detailed airflow in large spaces to algebraic multi-zone infiltration models. PMID- 12756001 TI - Impact of varying area of polluting surface materials on perceived air quality. AB - A laboratory study was performed to investigate the impact of the concentration of pollutants in the air on emissions from building materials. Building materials were placed in ventilated test chambers. The experimental set-up allowed the concentration of pollution in the exhaust air to be changed either by diluting exhaust air with clean air (changing the dilution factor) or by varying the area of the material inside the chamber when keeping the ventilation rate constant (changing the area factor). Four different building materials and three combinations of two or three building materials were studied in ventilated small scale test chambers. Each individual material and three of their combinations were examined at four different dilution factors and four different area factors. An untrained panel of 23 subjects assessed the air quality from the chambers. The results show that a certain increase in dilution improves the perceived air quality more than a similar decrease in area. The reason for this may be that the emission rate of odorous pollutants increases when the concentration in the chamber decreases. The results demonstrate that, in some cases the effect of increased ventilation on the air quality may be less than expected from a simple dilution model. PMID- 12756002 TI - House dust-mite allergen and cat allergen variability within carpeted living room floors in domestic dwellings. AB - Exposure to allergens from house dust-mites (Der p 1) and domestic cats (Fel d 1) is associated with symptom severity in atopic subjects with asthma and rhinitis. Assessment of allergen exposure in the domestic environment is normally determined by measurement from a single floor site. We determined the variability of these allergens and protein throughout the whole living room floor area. Dust samples were collected from 1 m2 areas from 16 carpeted living room floors in Wellington, New Zealand, and analyzed for concentrations of Der p 1 and Fel d 1. Mean coefficients of variation for Der p 1 and Fel d 1 were 53.1% (range: 28.5 136.8) and 65.6% (range: 28.5-131.0), respectively. This study has demonstrated a large variation of house dust-mite and cat allergens within living room floors and thus assessment of a single sampling site may not be representative of an individual's exposure risk. House dust-mite and cat allergen levels from the center of the room, in front of a couch or chair, or from a corner of the room are similar to mean levels from the whole room. These sites may thus be representative of the whole living room floor in large-scale epidemiological studies. PMID- 12756003 TI - Control strategies for sub-micrometer particles indoors: model study of air filtration and ventilation. AB - The effects of air filtration and ventilation on indoor particles were investigated using a single-zone mathematical model. Particle concentration indoors was predicted for several I/O conditions representing scenarios likely to occur in naturally and mechanically ventilated buildings. The effects were studied for static and dynamic conditions in a hypothetical office building. The input parameters were based on real-world data. For conditions with high particle concentrations outdoors, it is recommended to reduce the amount of outdoor air delivered indoors and the necessary reduction level can be quantified by the model simulation. Consideration should also be given to the thermal comfort and minimum outdoor air required for occupants. For conditions dominated by an indoor source, it is recommended to increase the amount of outdoor air delivered indoors and to reduce the amount of return air. Air filtration and ventilation reduce particle concentrations indoors, with the overall effect depending on efficiency, location and the number of filters applied. The assessment of indoor air quality for specific conditions could be easily calculated by the model using user defined input parameters. PMID- 12756004 TI - A screening assessment of emissions of volatile organic compounds and particles from heated indoor dust samples. AB - This paper characterizes and compares emissions during heating of different dust samples relevant to the indoor environment. Characterization includes emission of volatile organic compounds when dust samples were heated to 150 and 250 degrees C (gas chromatograph-mass spectrometer), weight loss during heating to 450 degrees C (thermogravimetric analysis), and the number of particles emitted during heating towards 200 degrees C (condensation nucleus counting). Element analyses were performed for non-heated dust (inductively coupled plasma discharge instrument). Emissions of volatile organic compounds from heated dust from different sources were surprisingly similar. However, the temperature at which the emission of volatiles started varied with the dust source. For most of the samples studied, the emissions were considerable already at 150 degrees C, and increased in number of peaks and peak area at 250 degrees C. Particle emissions started around 70 degrees C regardless of the dust source. Particle emissions seemed to be affected by the content of organic material. PMID- 12756005 TI - Modeling ventilation and radon in new Dutch dwellings. AB - Indoor radon concentrations were estimated for various ventilation conditions, the differences being mainly related to the airtightness of the dwelling and the ventilation behavior of its occupants. The estimations were aimed at describing the variation in air change rates and radon concentrations to be expected in the representative newly built Dutch dwellings and identifying the most important parameters determining air change rate and indoor radon concentration. The model estimations were compared with measurements. Most of the air was predicted to enter the model dwelling through leaks in the building shell, independent of the ventilation conditions of the dwelling. Opening the air inlets was shown to be an efficient way to increase infiltration and thus to decrease radon concentration. The effect of increasing the mechanical ventilation rate was considerably less than opening the air inlets. The mechanical ventilation sets the lower limit to the air change rate of the dwelling, and is effective in reducing the radon concentration when natural infiltration is low. Opening inside doors proved to be effective in preventing peak concentrations in poorly ventilated rooms. As the airtightness of newly built dwellings is still being improved, higher radon concentrations are to be expected in the near future and the effect of occupant behavior on indoor radon concentrations is likely to increase. According to the model estimations soil-borne radon played a moderate role, which is in line with measurements. PMID- 12756006 TI - Determinants of endotoxin levels in carpets in New Zealand homes. AB - Endotoxin in house dust has been shown to be associated with asthma severity. Little is known about the influence of housing characteristics on endotoxin distribution. Using standardized methods, dust was sampled from a 1m(2) site and the whole accessible carpet area in selected Wellington, New Zealand homes (n = 77). Endotoxin was measured using a Limulus Amoebocyte Lysate assay. Relative humidity and temperature were recorded using sensors placed in carpet bases. Questionnaires were used to collect information on housing characteristics. All analyses were performed for endotoxin units (EU)/mg and EU/m2 for each site. Geometric mean endotoxin levels were 22.7 EU/mg [geometric standard deviation (GSD) = 2.4] or 30,544 EU/m2 (GSD = 3.2) from the 1m(2) site, and 28.4 EU/mg (GSD = 3.4) or 5653 EU/m2 (GSD = 6.4) from the whole room. After controlling for confounding, endotoxin was positively associated with dogs inside [geometric mean ratio (GMR): 0.9-2.0], total household occupants (GMR: 1.7-2.0, for 1 m2 sample only), vacuum cleaners <1-year old (GMR: 2.3-2.7), reusing vacuum dust collection bags (GMR: 1.4-3.1), steamcleaning or shampooing the carpet (GMR: 1.4-2.2) and high relative humidity (GMR: 1.4-1.6). Lower endotoxin was associated with floor insulation (GMR: 0.4-0.8), and north-facing living rooms (GMR: 0.4-0.8). This study has identified home characteristics that could be modified to reduce endotoxin exposure. PMID- 12756008 TI - Determination of fungal spore release from wet building materials. AB - The release and transport of fungal spores from water-damaged building materials is a key factor for understanding the exposure to particles of fungal origin as a possible cause of adverse health effects associated to growth of fungi indoors. In this study, the release of spores from nine species of typical indoor fungi has been measured under controlled conditions. The fungi were cultivated for a period of 4-6 weeks on sterilized wet wallpapered gypsum boards at a relative humidity (RH) of approximately 97%. A specially designed small chamber (P-FLEC) was placed on the gypsum board. The release of fungal spores was induced by well defined jets of air impacting from rotating nozzles. The spores and other particles released from the surface were transported by the air flowing from the chamber through a top outlet to a particle counter and sizer. For two of the fungi (Penicillium chrysogenum and Trichoderma harzianum), the number of spores produced on the gypsum board and subsequently released was quantified. Also the relationship between air velocities from 0.3 to 3 m/s over the surface and spore release has been measured. The method was found to give very reproducible results for each fungal isolate, whereas the spore release is very different for different fungi under identical conditions. Also, the relationship between air velocity and spore release depends on the fungus. For some fungi a significant number of particles smaller than the spore size were released. The method applied in the study may also be useful for field studies and for generation of spores for exposure studies. PMID- 12756007 TI - The 3-year follow-up study in a block of flats - experiences in the use of the Finnish indoor climate classification. AB - Indoor climate of two new blocks of flats was investigated. The case building was built for people with respiratory diseases by following the instructions of the Finnish Classification of Indoor Climate, Construction and Finishing Materials, while the control building was built using conventional building technology. The main indoor air parameters (temperature, relative humidity and levels of CO, CO2, ammonia, total volatile organic compounds, total suspended particles, fungal spores, bacteria and cat, dog and house dust mite allergens) were measured in six apartments of both the buildings on five occasions during the 3-year occupancy. In addition, a questionnaire to evaluate symptoms of the occupants and their satisfaction with their home environment was conducted in connection with indoor air quality (IAQ) measurements. The levels of indoor air pollutants in the case building were, in general, lower than those in the control building. In addition, the asthmatic occupants informed that their symptoms had decreased during the occupancy in the case building. This case study showed that high IAQ is possible to reach by careful design, proper materials and equipment and on high-quality construction with reasonable additional costs. In addition, the study indicated that good IAQ can also be maintained during the occupancy, if sufficient information on factors affecting IAQ and guidance on proper use and care of equipment are available for occupants. PMID- 12756009 TI - Comparison of three small chamber test methods for the measurement of VOC emission rates from paint. AB - The aim of this study was to demonstrate a correlation between the measurement of emission rates of volatile organic compounds (VOCs) in three different climate chambers. In order to achieve this aim, the early state of the emission process in the three chambers was investigated and the effects of some important factors on the emission rates from paint were determined. The paper presents results of measurements in three different climate chambers. For the study, a 1-m3 chamber, a field and laboratory emission cell (FLEC), and a chamber for laboratory investigation of materials pollution and air quality (CLIMPAQ) were used. The airflow and surface area were selected so that the area-specific ventilation rates were identical in the three chambers. Temperature and relative humidity were identical during all the measurements. The paint examined was a solvent based alkyd paint intended for indoor, which use contained between 30 and 60% of white spirit in wet condition. The paint was applied to electropolished and cleaned stainless steel plates. After application, the test material was stored for 14 days for drying in a well-ventilated conditioning room before the measurements were made. After 2 weeks storage, the most pronounced emissions were pentanal, hexanal, octanal, and decanol. The period before the emission rate stabilized differed for the three chambers studied. However, all chambers gave similar emission rates within the overall uncertainty used in these experiments. PMID- 12756010 TI - Characterization of endotoxin and 3-hydroxy fatty acid levels in air and settled dust from commercial aircraft cabins. AB - Endotoxin was measured in air and dust samples collected during four commercial aircraft flights. Samples were analyzed for endotoxin biological activity using the Limulus assay. 3-hydroxy fatty acids (3-OH FA) of carbon chain lengths C10:0 C18:0 were determined in dust by gas chromatography-ion trap tandem mass spectrometry. The geometric mean (geometric standard deviation) endotoxin air level was 1.5 EU/m3 (1.9, n = 28); however, significant differences were found by flight within aircraft type. Mean endotoxin levels were significantly higher in carpet dust than in seat dust (140 +/- 81 vs. 51 +/- 25 EU/mg dust, n = 32 each, P < 0.001). Airborne endotoxin levels were not significantly related to either carpet or seat dust endotoxin levels. Mean 3-OH FA levels were significantly higher in carpet dust than in seat dust for C10:2, C12:0, and C14:0 (P < 0.001 for each), while the mean level of C16:0 was significantly higher in seat dust than in carpet dust (P < 0.01). Carpet dust endotoxin was significantly, but moderately, correlated with 3-OH-C12:0 and 3-OH-C14:0 (Pearson r = 0.52 and 0.48, respectively), while correlation of seat dust endotoxin with individual 3-OH FAs depended on the test statistic used. Mean endotoxin potency was significantly higher for carpet dust than for seat dust (6.3 +/- 3.0 vs. 3.0 +/- 1.4 EU/pmol LPS, P < 0.0001). Mean endotoxin levels in the air and dust of commercial aircraft cabins were generally higher than mean levels reported in homes and office buildings. These results suggest that exposure route and dust source are important considerations when relating endotoxin exposure to specific health outcomes. PMID- 12756011 TI - How do the indoor size distributions of airborne submicron and ultrafine particles in the absence of significant indoor sources depend on outdoor distributions? AB - Although almost all epidemiological studies of smaller airborne particles only consider outdoor concentrations, people in Central Europe actually spend most of their time indoors. Yet indoor pollutants such as organic gases, allergens and dust are known to play a prominent role, often affecting human health more than outdoor ones. The aim of this study was to ascertain how the indoor particle size distributions of submicron and ultrafine particles correlate with the outdoor concentrations in the absence of significant indoor sources. A typical indoor particle size distribution pattern has one or two modes. In the absence of significant indoor activities such as smoking, cooking etc., outdoor particles were found to be a very important source of indoor particles. The study shows that in the absence of significant indoor sources, the number of indoor concentrations of particles in this size range are clearly lower than the outdoor concentrations. This difference is greater, the higher the number of outdoor concentrations. However, the drop in concentration is not uniform, with the decrease in concentration of smaller particles exceeding that of larger ones. By contrast, the findings with larger particle sizes (diameter > 1 microm) exhibit rather linear concentration decreases. The non-uniform drop in the number of concentrations from outdoors to indoors in our measurements considering smaller particles ( >0.01 microm) is accompanied by a shift of the concentration maxima to larger particle diameters. PMID- 12756012 TI - Analysis of visitation frequency through particle tracking method based on LES and model experiment. AB - As ventilation efficiency in a room is not always uniformly distributed, an index for measuring ventilation efficiency at a concerned point or in a concerned local domain is required. Local ventilation efficiency is often represented by the rate of the averaged concentration of the local domain to that of exhausted air from the room. From the age theory of air, it is well known that the averaged concentration in a room corresponds to the mean staying time of contaminant. Evaluating the domain-averaged concentration (Cdomain) means evaluating the average staying time of the contaminant in the domain. It can be only one part of the whole room and can be considered as an occupied zone. Visitation frequency (VF) and the average staying time of the contaminant for one visitation in the local domain (Tp) are introduced to analyze the average staying time of the contaminant in the local domain. The value of VF is strongly affected by the position of the local domain in the room; that is, the position of the local domain in the whole flow field of the room. Tp represents the property of the flow pattern in the local domain. As the indication of VF and Tp represent the mechanism for forming the domain-averaged concentration, they are deeply related to local purging flow rate, which represents the airflow rate for defining the domain-averaged concentration. As VF and Tp are related to the contaminant transportation property, it is effective to analyze them by particle tracking method. A CFD method of large eddy simulation (LES) was thereby carried out in this study. The prediction result by LES is also validated by a precise model experiment. In this paper, the detailed analysis of VF and Tp is carried out on the basis of the particle tracking method utilizing the LES result in order to clarify the mechanism of the domain-averaged concentration. The analyzed room has one supply inlet and one exhaust outlet. A clear re-circulating flow, generated by the forced ventilation, is observed in the room. The value of VF is examined with three types of local domains in the room model. In the room model, VF shows a value of 5.70 when the local domain occupies half of the room. It becomes smaller when the size of the local domain is reduced. PMID- 12756013 TI - Contribution of incense burning to indoor PM10 and particle-bound polycyclic aromatic hydrocarbons under two ventilation conditions. AB - Burning incense to worship Gods and ancestors is a traditional practice prevalent in Asian societies. This work investigated indoor PM10 concentrations resulting from incense burning in household environments under two conditions: closed and ventilated. The exposure concentrations of particle-bound polycyclic aromatic hydrocarbons (PAHs) were estimated. The factors of potential exposure were also evaluated. Under both conditions, samples were taken at three locations: 0.3, 3.5 and 7 m away from the altar during three periods: incense burning, the first 3 h, and the 4-6 h after cessation of combustion. PAH concentrations of incense smoke were assessed in the laboratory. Personal environment monitors were used as sampling instruments. The results showed a significant contribution of incense burning to indoor PM10 and particulate PAH concentrations. PM10 concentrations near the altar during incense burning were 723 and 178 microg/m3, more than nine and 1.6 times background levels, under closed and ventilated conditions, respectively. Exposure concentrations of particle-bound PAHs were 0.088-0.45 microg/m3 during incense burning. On average, PM10 and associated PAH concentrations were about 371 and 0.23 microg/m3 lower, respectively, in ventilated environments compared with closed conditions. Concentrations were elevated for at least 6 h under closed conditions. PMID- 12756014 TI - Air flow and particle control with different ventilation systems in a classroom. AB - Most ventilation and air conditioning systems are designed without much concern about how settling particles behave in ventilation air flows. For displacement ventilation systems, designers normally assume that all pollutants follow the buoyant air flow into an upper zone, where they are evacuated. This is, however, not always true. Previous studies show that high concentrations of settling respirable particles can be found in the breathing zone, and that the exposure rates can be a health hazard to occupants. The emphasis here is on how ventilation systems should be designed to minimize respirable airborne particles in the breathing zone. The supply and exhaust conditions of the ventilation air flow are shown to play an important role in the control of air quality. Computer simulation programs of computational fluid dynamics (CFD) type are used. Particle concentrations, thermal conditions and modified ventilation system solutions are reported. PMID- 12756015 TI - Investigation of megakaryocyte apoptosis in children with acute and chronic idiopathic thrombocytopenic purpura. AB - OBJECTIVE: Although the platelet destruction shows a primary role in the thrombocytopenia of idiopathic thrombocytopenic purpura (ITP), it has been demonstrated that impaired platelet production may also contribute to the severity of thrombocytopenia in ITP. The present study examined megakaryocyte apoptosis in bone marrow aspirates of children with acute and chronic ITP and investigated the role of megakaryocyte apoptosis in ITP pathophysiology. METHODS: Thirteen children diagnosed with acute ITP and eight children diagnosed with chronic ITP comprised the study group. Ten children, who were hospitalized for scoliosis operation but healthy otherwise, comprised the control group. In all children, megakaryocytes were isolated from the same amount of bone marrow aspirate samples using MACS CD61 MicroBeads (Miltenyl Biotec, Auburn, CA, USA). Megakaryocyte apoptosis was studied with transferase-mediated d-UTP-bitin nick end-labeling method. RESULTS: Isolated megakaryocyte counts did not differ significantly between acute ITP, chronic ITP and control groups. The percentage of apoptotic megakaryocytes did not differ significantly between acute ITP group and control group and between chronic ITP group and control group. The percentage of apoptotic megakaryocytes in patients with chronic ITP was significantly lower than the patients with acute ITP. There was no correlation between the percentage of apoptotic megakaryocytes and platelet counts of the cases. CONCLUSIONS: Increased megakaryocytic apoptosis does not play a role in the pathogenesis of dysmegakaryopoiesis and impaired platelet production in children with ITP. Decreased megakaryocyte apoptosis in cases with chronic ITP may be due to suppression of megakaryocyte maturation, as the terminal phase of the megakaryocyte lifespan is characterized by the onset of apoptosis. PMID- 12756016 TI - Mycophenolate mofetil (MMF) for the treatment of steroid-resistant idiopathic thrombocytopenic purpura. AB - The treatment of chronic idiopathic thrombocytopenic purpura (ITP) is difficult in those unresponsive to corticosteroids and/or splenectomy. We attempted to induce durable response in 21 patients with refractory ITP by applying mycophenolate mofetil (MMF) (1.5-2.0 g/d), a novel immunosuppressive agent. Overall response rate was 62% (13 of 21), including 24% (five of 21) in complete response (CR), 29% (six of 21) in partial response (PR), and 10% (two of 21) in minor response (MR). The response rates for non-splenectomized and splenectomized ITP patients were 64% (nine of 14) and 57% (four of seven), respectively (P > 0.05). 39% (five of 13) responders relapsed as a result of dose reduction or withdraw of MMF, and 61% (eight of 13) responders maintained their effectiveness for a median of 24 wk. Sustained response was observed in three patients in whom MMF was withdrawn. MMF was well tolerated with only slight nausea and diarrhea recorded in 3 of 21 cases. No premature withdrawal was found in this study. CD3+ peripheral blood mononuclear cells (PBMC) and CD19+ PBMC were significantly reduced 12 wk after MMF administration in the responders. Platelet-associated antibodies against glycoproteins GPIIb/IIIa were detected in 13 of 21 (62%) patients before MMF treatment, and antibody levels were significantly decreased in responders 12 wk after MMF administration. This suggested that MMF might correct the immunologic abnormalities underlying the destruction of circulating platelets in ITP. We conclude that MMF could be used as a second-line agent for the treatment of steroid-resistant ITP before or after splenectomy and thereby is worth of further evaluation in randomized studies. PMID- 12756017 TI - The presence of a significant association between elevated PRV-1 mRNA expression and low plasma erythropoietin concentration in essential thrombocythaemia. AB - Approximately 45% of newly diagnosed patients with essential thrombocythaemia (ET) demonstrate subnormal plasma erythropoietin (EPO) concentrations, which constitutes a risk factor for occlusive vascular events. In 58 ET patients, a possible association between polycythaemia rubra vera-1 (PRV-1) overexpression and subnormal plasma EPO was investigated, which was always measured prior to the institution of platelet lowering agents. At the time when PRV-1 expression was measured, 28 of 58 (48%) ET patients had received platelet lowering treatment. PRV-1 expression was measured by quantitative real-time reverse transcription polymerase chain reaction assay of mRNA extracted from purified peripheral blood buffy coat. The cycle threshold (CT) value of PRV-1 was determined and was divided with the CT value for the housekeeping GAPDH gene transcript. A quotient <0.93 was defined as PRV-1 positive. Of the ET patients 12 of 58 (21%) were PRV-1 positive and 19 of 58 (33%) demonstrated subnormal plasma EPO. In the 58 ET patients there was a significant association between low plasma EPO and PRV-1 positive results (P = 0.001). The 30 ET patients who had not received any platelet lowering treatment showed a significant (P = 0.005) relation between PRV 1 positivity and subnormal plasma EPO. No such relationship was present in the 28 ET patients who had received prior treatment with the above drugs (P = 0.147). PMID- 12756018 TI - High-resolution comparative genomic hybridisation yields a high detection rate of chromosomal aberrations in childhood acute lymphoblastic leukaemia. AB - BACKGROUND: Cytogenetic aberrations are of prognostic significance in childhood acute lymphoblastic leukaemias and a high detection rate could improve the biological understanding and classification of these diseases. METHODS: Bone marrow samples from 92 children with acute lymphoblastic leukaemia were studied by high-resolution comparative genomic hybridisation (HRCGH) using dynamic standard reference intervals that enhance both specificity and sensitivity in the detection of aberrations. RESULTS: In 80 patients (87%) HRCGH revealed a total of 405 aberrations, mostly whole chromosome gains (n = 265) and partial losses (n = 80). The 25 leukaemias with a gain of more than five whole chromosomes by HRCGH harboured only 7% of all losses. With G-band karyotyping 59 patients (64%) had aberrations. HRCGH revealed more aberrations per patient than did G-band karyotyping (median: 3 vs. 1, P = 0.005), revealed aberrations in 27 of the 34 patients for whom the G-band karyotyping failed or was found to be normal, and specifically revealed more 9p losses (21% vs. 5%, P < 0.005), 12p losses (12% vs. 2%, P < 0.05) and 17q gains (11% vs. 1%, P < 0.01). Compared to the present study, the frequency of patients with aberrant karyotypes was significantly lower in previous conventional CGH studies (64% vs. 87%, P < 0.0001), as was the rate of partial aberrations per patient (1.1% vs. 1.7, P < 0.001), particularly with fewer 6q losses, 9p losses and 17q gains detected. CONCLUSION: HRCGH is superior to conventional CGH as an adjunct to G-band karyotyping as it detects recurrent aberrations at a significantly higher rate than both these techniques. PMID- 12756019 TI - Increased serum levels of matrix metalloproteinase-9 predict clinical outcome of patients with early B-cell chronic lymphocytic leukaemia. AB - BACKGROUND AND METHODS: Serum levels of matrix metalloproteinase-9 (MMP-9) which agree with progression in solid and haematological tumours were correlated to the risk of disease progression in 62 patients with early (Binet stage A) B-cell chronic lymphocytic leukaemia (CLL). Sera were taken at diagnosis and tested by an enzyme-linked immunosorbent assay. RESULTS: MMP-9 levels positively correlated with haemoglobin levels (P = 0.03) and platelet count (P = 0.03). No association was found with main clinico-haematological features representative of tumour mass, such as peripheral blood lymphocytosis, bone marrow histology, Rai substages and beta-2 microglobulin (beta-2m). A cut-off of MMP-9 levels corresponding to 33rd percentile (203 ng/mL) or higher identified earlier upstaging and shorter progression-free survival. MMP-9 was a significant prognostic marker in multivariate analysis and partially independent of Rai substages, which suggests its inclusion into such a staging system to better stratify prognostically Rai stages I and II patients. CONCLUSIONS: MMP-9 serum levels predict disease behaviour and help to refine the prognosis of stage A CLL patients. PMID- 12756020 TI - Feasibility of fludarabine added to VAD during induction therapy in multiple myeloma: a randomised phase II-study. AB - Multiple myeloma (MM) is considered to be an essentially incurable haematological malignant disease, probably because of the existence of resistant clonal precursor cell with self-renewal capacity. Recent data have indicated that the myeloma cell hierarchy includes circulating clonal memory B cells, which differ considerably from the classical end-stage plasma cells, infiltrating the bone marrow. The pathophysiological significance of these cells is unknown, but hypothetically they may serve as 'sleeping' myeloma stem cells responsible for and 'feeding' post-treatment relapse and progression. The present study evaluates the toxicity and feasibility of fludarabine, added to the VAD-induction regimen in MM, and investigates the effect on the myeloma cell hierarchy. Nineteen patients were randomised to receive either four cycles of VAD (n = 9) or two cycles of VAD, followed by two cycles of VAD combined with 5 days fludarabine 25 mg/m2/day i.v. (n = 10). Toxicity evaluation showed more profound neutropenia in the fludarabine-treated patients and two infectious episodes in each study arm: three were fever of unknown origin while one, in the fludarabine-arm, was a local skin infection at the insertion site of the central venous line. Nine of the fludarabine-treated patients responded to treatment (two complete remission, seven partial remission), compared with five responders (all PR) in the control arm. The effects on the blood circulating myeloma compartments identified an increased reduction of CD19+ B cells and myeloma plasma cells in the fludarabine arm. In conclusion, adding fludarabine to VAD induction in multiple myeloma is feasible and may be clinically effective by reducing the myeloma clone. PMID- 12756021 TI - Serum levels of cytokines correlate to donor chimerism and acute graft-vs.-host disease after haematopoietic stem cell transplantation. AB - BACKGROUND: Some patients become full donor chimeras (DC) early after stem-cell transplantation (SCT), while others remain mixed chimeras for a longer time. Little is known about the mechanism behind these phenomena. METHODS: Serum cytokine levels during conditioning and during the first month after SCT were analysed in 30 patients. Of the 21 patients who became full T-cell DC from the first analysed sample, 12 developed grade II-IV acute graft-vs.-host disease (GVHD) and the other nine, mild or no acute GVHD. Another nine patients were T cell mixed chimeras (MC). All MC patients had no or mild acute GVHD. RESULTS: During the pretransplant conditioning, DC patients had higher levels of tumour necrosis factor (TNF)-alpha and lower levels of transforming growth factor (TGF) beta and interleukin (IL)-10, compared with MC patients. During the first week after SCT, lower levels of TGF-beta and IL-10 and higher levels of soluble Fas (sFas) were found in DC patients compared with MC patients. During the second and third weeks after SCT, increased levels of TNF-alpha, interferon (IFN)-gamma and sFas were found among DC patients compared with MC patients. Patients who developed moderate-to-severe acute GVHD had higher levels of TNF-alpha, IFN gamma, IL-10 and sFas at 2 weeks post-SCT than in those with less GVHD. Patients homozygous for the TNFd microsatellite alleles 3 or 4 had significantly higher TNF-alpha levels during conditioning and more often developed acute GVHD grades II-IV. CONCLUSION: These results indicate that an imbalance between pro inflammatory and immune- modulating cytokines are involved in the development of chimerism and acute GVHD after allo-SCT. The Fas/FasL pathway is probably involved in the elimination of recipient cells leading to full donor chimerism. PMID- 12756022 TI - Safety monitoring of cardiac and hepatic systems in beta-thalassemia patients with chelating treatment in Taiwan. AB - We conducted a prospective 3-yr clinical study comparing deferiprone (L1) with desferrioxamine (DFX). The therapeutic efficacy and potential side-effects on cardiac and/or hepatic systems of thalassemia patients were assessed by left ventricular ejection fraction, T2-weighted magnetic resonance imaging, and biochemical parameters. In both groups, levels of serum ferritin decreased significantly, and the hepatic function improved notably. Besides decrement of iron, no marked pathohistological changes were observed in the liver biopsies. These results indicated that for patients who failed to respond to DFX treatment, the use of L1 to remove excess iron deposition is recommended. PMID- 12756023 TI - Reversal of cardiac complications in thalassemia major by long-term intermittent daily intensive iron chelation. AB - OBJECTIVES: In patients with thalassemia major (TM) who are non-compliant with long-term deferoxamine (DFO) chelation, survival is limited mainly because of cardiac complications of transfusional siderosis. It was recently shown in a small group of TM patients with established cardiac damage that continuous 24-h DFO infusion via an indwelling intravenous (i.v.) catheter is effective in reversing cardiac toxicity. The aim of the present study was to evaluate the results with intermittent daily (8-10 h) i.v. DFO. PATIENTS: Eight TM patients with cardiac complications treated with intensive intermittent DFO were retrospectively evaluated by the mean annual serum ferritin, radionucleated ventriculography and 24-h electrocardiography recordings. RESULTS: The median age at diagnosis of cardiac disease was 17.5 yr (range 14-21), and the median follow up time was 84 months (range, 36-120). In the majority of patients (seven of eight) high-dose DFO (mean 95 +/- 18.3 mg/kg/d) was administered via a central venous line. During follow-up, there was a significant decrease in the mean ferritin levels (5828 +/- 2016 ng/mL to 1585 +/- 1849 ng/mL, P < 0.001). Both cardiac failure (mean ejection fraction 32 +/- 5) and cardiac arrhythmias were resolved in four of five patients. One non-compliant patient died during the follow-up. Following discontinuation of the i.v. therapy, compliance with conventional DFO therapy improved. The complications of this regimen, mainly catheter-related infections and catheter-related thrombosis, were similar to those described earlier. CONCLUSIONS: These results with the longest follow-up period in the literature suggest that i.v. high-dose DFO for 8-10 h daily may be as effective as continuous 24-h infusion for the reversal of established cardiac disease in TM. PMID- 12756024 TI - Compound heterozygosity of two missense mutations in the NADH-cytochrome b5 reductase gene of a Polish patient with type I recessive congenital methaemoglobinaemia. AB - A case of type I methaemoglobinaemia observed in a Polish subject with compound heterozygosity for two mutations in the reduced nicotinamide adenine dinucleotide (NADH) cytochrome b5 reductase (b5R) gene is described. One is a novel mutation 647T-->C which leads to substitution of isoleucine by threonine at position 215 (I215T). This maternal mutation was found in several family members. A previously known mutation, 757G-->A, leads to the replacement of valine by methionine at position 252 (V252M). The latter mutation was found also in the father and one of the two brothers. The effects of these mutations were analysed on a model of the human b5R protein obtained by homology modelling. Although both amino acid substitutions are located in the NADH-binding domain, the whole protein structure, especially the region between the flavin adenine dinucleotide and NADH binding domains, is disturbed. The structural changes in the I215T mutant are less prominent than those in the V252M mutant. We presume that the 647T-->C mutation is a type I mutation, however, it has not been observed in the homozygous state. PMID- 12756025 TI - Haematuria and urolithiasis in patients with haemophilia. AB - Recurrent haematuria was present in 18 of 474 moderate and severe haemophiliacs, the cause of which was found to be urolithiasis in six patients (33%). The prevalence of urolithiasis in haemophiliacs was found to be significantly higher than that reported from the general population, i.e. 4.5 of 10 000 population under 40 yr of age (odds ratio (OR) 23.4; 95% CI 18.2-28.7; lambda2 test P < 0.01). Even when this prevalence was corrected for gender bias, i.e. male : female (5 : 1), the significance of the present findings remain (OR 17.6; 95% CI 13.8-21.5; lambda2 test P < 0.01). In developing countries, severe and moderately severe haemophiliacs may be at a higher risk of urolithiasis because of prolonged recumbency necessitated by recurrent joint bleeds and inadequate replacement therapy. PMID- 12756026 TI - Successful treatment of a patient with subcutaneous panniculitis-like T-cell lymphoma with high-dose chemotherapy and total body irradiation. AB - A 24-yr-old man was referred for fever, right cheek swelling, subcutaneous tumor and liver dysfunction. Physical examination showed an elastic hard subcutaneous tumor on the right cheek, left axillary lymph node swelling and multiple small subcutaneous tumors in the trunk. Laboratory examinations showed elevated levels of transaminase, soluble interleukin-2 receptor and ferritin. Biopsy of the subcutaneous tumor showed proliferation of medium-sized cells with abundant clear cytoplasm and hyperchromatic nuclei among the subcutaneous fat tissues. These cells showed CD3+, CD4-, CD8+, CD56- and CD20- phenotype and possessed cytotoxic molecules such as granzyme B and T-cell intracellular antigen-1. Bone marrow aspiration showed proliferation of small numbers of abnormal lymphocytes with severe hemophagocytosis. He was thus diagnosed as having subcutaneous panniculitis-like T-cell lymphoma (SPTCL) and treated with dose-escalated CHOP regimen. After three courses of the chemotherapy, he was further treated with high-dose chemotherapy and total body irradiation (TBI) with autologous peripheral blood stem cell rescue. Thereafter, he has been in remission for more than 2 yr. We consider that SPTCL with hemophagocytosis is an extremely aggressive disease, and high-dose chemotherapy and TBI should be included for the choice of the treatment. PMID- 12756027 TI - RANKL expression in a case of follicular lymphoma. AB - The TNF-family molecule, RANKL, is a key regulator of bone remodeling and essential for the development and activation of osteoclasis. Bone involvement signals diesease activity in non-Hodgkin's lymphoma and influences the progenesis. The molecular mechanism and soluble factors involved in osteoclastic activation in haematological malignancies remain unclear except for Multiple Myeloma and Adult T-cell Leukemia. The aim of this paper is to report the first case of Follicular Lymphoma with bone involvement displaying an aberrant expression of RANKL in malignant cells. The detection of RANKL in Follicullar Lymphoma may help to prevent bone lesion in patients by determining an appropriate treatment. PMID- 12756028 TI - Bull's eye maculopathy with deferoxamine. PMID- 12756029 TI - A profile of the patient with periodontal disease? PMID- 12756030 TI - Understanding the etiology of periodontitis: an overview of periodontal risk factors. PMID- 12756031 TI - Microorganisms as risk indicators for periodontal disease. PMID- 12756032 TI - The genetic relationship to periodontal disease. PMID- 12756033 TI - Smoking and periodontal disease. PMID- 12756034 TI - Hormonal influences: effects of diabetes mellitus and endogenous female sex steroid hormones on the periodontium. PMID- 12756035 TI - Systemic disease and periodontitis: manifestations of neutrophil dysfunction. PMID- 12756036 TI - Osteoporosis and periodontal disease progression. PMID- 12756037 TI - Occlusal forces as a risk factor for periodontal disease. PMID- 12756038 TI - Acquired immune suppression and other risk factors/indicators for periodontal disease progression. PMID- 12756039 TI - Kidney transplants for children under 1 year of age - a single-center experience. AB - From September 20, 1970 to October 24, 2001, we performed 46 kidney transplants in infants under 1 yr old at the University of Minnesota. This article reviews the preoperative care, surgical technique, and immunosuppression. Recipients included 16 females and 30 males; the youngest recipient was 6 wk old. The mean pretransplant height was 62.8 cm, which increased to 77 cm at 1 yr post transplant and to 104 cm at 5 yr. We used 40 living donors (all but 1 were related to the recipient) and 6 cadaver donors. The overall actuarial graft survival was 85% at 1 yr and 70% at 5 yr. In the cyclosporine era, graft survival improved to 91% at 1 yr and 80% at 5 yr. Death with function was the most common cause of graft loss (n = 5), followed by biopsy-proven chronic rejection (n = 4), biopsy-proven recurrent disease (n = 3), and graft thrombosis (n = 2). Patient survival was 91% at 1 yr and 86% at 5 yr. In the cyclosporine era, patient survival was 100% at 5 yr and 85% at 10 yr. We concluded that an early transplant is the best treatment option for infants under 1 yr old with chronic renal failure. Whenever possible, adult living kidney donors should be used. PMID- 12756040 TI - Reduced-intensity allogeneic hematopoietic cell transplantation: Graft versus tumor effects with decreased toxicity. AB - The potentially curative role of allogeneic hematopoietic cell transplantation (HCT) in neoplastic and non-neoplastic diseases is offset by the substantial risks of morbidity and mortality from complications of the intensive myeloablative and immunosuppressive preparative regimen. These regimen-related toxicities have restricted allogeneic HCT to young, otherwise healthy individuals without comorbid diseases. Pediatric patients undergoing conventional allogeneic HCT have lower procedure-related mortality but are at risk for non-fatal late effects of the high-dose pretransplant chemoradiotherapy, such as growth retardation, sterility and other endocrine dysfunction. Evaluation of reduced intensity preparative regimens is the major focus of current clinical research in allogeneic HCT. Reduced-intensity HCT (RI-HCT) relies on the use of immunosuppressive but non-myeloablative agents that allow engraftment of donor cells, which provide adoptive allogeneic cellular immunotherapy and graft versus tumor (GVT) effects, with decreased regimen-related toxicities. Although the experience with RI-HCT in pediatric patients is very limited at this time, results in adults indicate that attenuated-dose preparative regimens allow older patients and those with organ dysfunction to undergo successful allogeneic HCT with acceptable morbidity and mortality. In adults, the potency of the allogeneic GVT effect varies among neoplastic diseases, with better results observed in patients with indolent hematological malignancies or renal cell carcinoma. The effectiveness of RI-HCT as treatment for children with hemoglobinopathies, chronic granulomatous disease and cellular immunodeficiencies is encouraging, and the role of reduced-intensity preparative regimens for allogeneic HCT in pediatric malignancies is under investigation. PMID- 12756041 TI - Pilot study describing the use of pravastatin in pediatric renal transplant recipients. AB - Renal transplant (Tx) recipients frequently develop hypercholesterolemia. Pravastatin (P) has been shown to be effective in adult renal Tx recipients, not only in reducing serum cholesterol, but possibly also in decreasing graft rejection. However, there are no data on the use of P in children following renal transplantation. We conducted a retrospective case-control study evaluating the safety and efficacy of P (10-20 mg/day) in reducing hypercholesterolemia, when used pre-emptively in the post-Tx period in seven children, compared with an historical control (C) group of nine children who had not received P. The two groups were comparable with respect to their demographics and in their pretransplant serum cholesterol. Compared with the C group, the mean serum cholesterol in the P group was lower at 3 months (159 mg/dL vs. 225 mg/dL), 6 months (134 mg/dL vs. 200 mg/dL), 9 months (134 mg/dL vs. 209 mg/dL), and 12 months (125 mg/dL vs. 195 mg/dL) (p < 0.005 for all, Student's two-tailed t test). At 1 month only 43% of the P group had hypercholesterolemia compared with 67% of the controls; by 12 months this difference was even more significant (0% in the P group vs. 45% in the C group). None of the treated patients developed any adverse reactions. This study demonstrates that the pre-emptive use of P in pediatric renal Tx recipients appears to be effective in significantly reducing serum cholesterol. Whether this effect will translate into improved allograft and patient survival in the long term cannot be predicted at present and will require additional studies to evaluate. PMID- 12756042 TI - Exfoliative rejection after intestinal transplantation in children. AB - BACKGROUND: Graft rejection is the most significant cause of allograft failure after intestinal transplantation (ITx). Severity can vary and is based on histologic criteria, the most extreme form being exfoliation of the mucosa. We present the characteristics and outcome of children who developed exfoliative rejection (ER) after ITx. METHODS: Between June 1990 and March 2002, 88 patients received 92 ITx which included isolated small bowel (SB, n = 26), combined liver small bowel (LSB, n = 54), and multivisceral MV n = 12) allografts performed under tacrolimus and steroid immunosuppresson. ER was diagnosed by endoscopy and confirmed by biopsy. RESULTS: Thirteen (15%) of 88 patients developed 15 episodes of ER in 15 intestinal allografts, and included SB (n = 8), LSB (n = 5), and MV (n = 2). Time to ER after ITx ranged from 9 days to 45.5 months (median 22 days). Eight episodes of ER developed within 1 month after ITx. Ten episodes of ER were exacerbations of prior rejection. Five episodes occurred abruptly. All but one received OKT3. Fourteen of 15 allografts were lost; six patients underwent allograft enterectomy acutely as a salvage operation because of ER. The remainder of the allografts were either removed or lost to patient death as a consequence to infection or chronic rejection after resolution of ER. Retransplantation was performed in three patients, with subsequent recurrence of ER in two retransplanted allografts. Inclusion of a liver allograft was a protective factor toward decreasing the incidence of ER. The results of cross-matching, inclusion of a colonic segment, and simultaneous bone marrow infusion did not affect the incidence of ER. Infectious complications included post-transplant lymphoproliferative disease (n = 4), cytomegalovirus (n = 5), and adenovirus infection (n = 2). CONCLUSIONS: Exfoliative rejection is associated with a high morbidity and mortality after ITx. Strategies to improve survival may include up front anti-lymphocyte antibody therapy and, when fail to respond promptly and satisfactorily, early intestinal allograft removal. PMID- 12756043 TI - Growth impairment at renal transplantation--a determinant of growth and final height. AB - Long-term outcome of growth and final adult height (FH) are a major concern of children after a renal transplantation (Tx). We therefore studied the yearly measurements of height (Ht), expressed as the Z-score and bone age (BA), in 58 children (28 girls) transplanted in our departments and followed-up for 5-18 (mean 9.6) yr after the operation. Twenty-four children reached final adult height. Renal function was evaluated as the glomerular filtration rate (GFR), which is estimated from the clearance of inulin. The mean Ht Z-score at Tx was 1.3 in girls and -2.7 in boys and increased to -0.6 and -1.5, respectively, at 5 yr after Tx. The greatest increase, seen in the shortest children and those transplanted before 7 yr of age, occurred during the first 3 yr. Children aged 7 12 yr at Tx showed an increase in height during the first years after the transplant, while those transplanted at >12 yr of age were not growth-retarded and therefore did not change their Ht Z-score. The most growth-retarded were also the most BA retarded. The mean FH Z-score was -1.1. A direct correlation was seen between GFR at 1 yr after Tx and the increase in height Z-score from Tx to 1 and 5 yr after Tx. In summary, the increment in height following Tx was the greatest in the most growth-retarded children and most marked during the first 3 yr after the transplant. FH was within normal range in 75% of the children. A high Ht Z score at Tx had a positive effect on FH. Thus, growth after Tx was affected by the degree of stunting at Tx and renal function after Tx. PMID- 12756044 TI - What are the best pulmonary function test parameters for early detection of post lung transplant bronchiolitis obliterans syndrome in children? AB - Post-lung transplant bronchiolitis obliterans syndrome (BOS) is defined as an unexplained fall in forced expiratory volume in 1 s (FEV1) >or=20% of baseline (B). There have been reports in adults that FEF25-75% (>30% decline from B) is more sensitive than FEV1 for the early diagnosis of BOS. Yet, it is not known if other pulmonary function test (PFT) parameters - forced expiratory flow rates at 25-75% of vital capacity (FEF25-75%) and maximal expiratory flow rate at 80% (Vmax80%), 70% (Vmax70%) and 60% (Vmax60%) - are more sensitive indicators for early diagnosis of BOS than FEV1 in post-lung transplant children. We reviewed serial PFTs of 18 patients (ages 14.1 +/- 3.7 yr, 50% female) who had lung transplantation at our institution from 1993 to 1999, and who met the criteria for BOS diagnosis. There was no significant difference in post-transplant days when decline in FEV1 >or=20% of B, FEF25-75% >30% of B, and Vmax80%, Vmax70% and Vmax60% from normal occurred (635 +/- 431, 551 +/- 422 and 454 +/- 287 days, respectively; p = 0.4). However, a decline in FEV1 was the first abnormality in only 39% of the patients, while a decline in FEF25-75% and Vmax at specific lung volume were the first abnormality in 78% and 56% of the patients, respectively. The earliest signs of BOS would be missed in 61% of patients if FEV1 was the primary parameter used for the diagnosis. In order to improve the sensitivity of the diagnosis of post-lung transplant BOS; we speculate that the diagnosis should be based on decreases in FEF25-75% rather than on FEV1. PMID- 12756045 TI - Pretransplant peritoneal dialysis and graft thrombosis following pediatric kidney transplantation: a NAPRTCS report. AB - Graft thrombosis is a common cause of graft failure in pediatric renal transplantation. Several previous studies, including a North American Pediatric Renal Transplant Cooperative Study (NAPRTCS) review of pretransplant dialysis status and graft outcomes, have described a potential correlation of peritoneal dialysis (PD) and graft thrombosis. This issue is of particular concern for pediatric transplant programs as more than 65% of children with end stage renal disease are treated with PD. We reviewed 7247 pediatric renal transplants performed between 1987 and 2001. Thrombosis was the cause of graft loss in 2.7% (199) of all the transplants performed. Among failed transplants, thrombosis was the third most common cause of graft loss in both index (11.6%) and subsequent transplants (14.5%). Thrombosis becomes the most common cause of graft failure (21%, 61/294) if one looks at transplants in the later cohort, from 1996 to 2001. This change is primarily because of a decrease in the incidence of acute rejection. In the PD group, 3.4% of all grafts were lost as a result of thrombosis. This compares with 1.9% in the hemodialysis group, 2.4% in the pre emptive transplant group, and 4.1% among patients who received both dialysis modalities. There was a statistically significant difference in thrombosis failure risk in the different dialysis groups (p = 0.005) with those who received only peritoneal dialysis having the highest risk. Additional significant risk factors for graft thrombosis included; cadaver donor source (p < 0.001), cold ischemia time >24 h (p < 0.001), history of prior transplant (p < 0.001), donor age <6 yr (p < 0.001), and >5 pretransplant blood transfusions (p = 0.02). Using stepwise proportional hazards modeling, only pretransplant peritoneal dialysis, >24 h cold ischemia time, prior transplant, and donor age <6 yr were simultaneously associated with an increased risk of thrombosis. We conclude that pretransplant PD is associated with an increased risk of graft thrombosis. Special precautions should be undertaken in pediatric renal transplant patients who have received PD, especially infants and young children. PMID- 12756046 TI - Does the transplantation process modify the immunogenicity of fetal adrenal grafts in rat? AB - The concept that fetal tissue transplants enjoy an immunologic privilege grounds on the primary immaturity of major histocompatibility complex (MHC) expression. However, experiences in human organ transplantation reveal that the immunogenicity of any graft could be modified by external factors such as ischemia. Consequently, the question arises, whether the process of transplantation modifies the immunogenicity of fetal grafts. In a syngeneic rat model (Lewis), fetal adrenal glands were transplanted into the greater omentum of adult hosts. After harvesting the grafts sequentially, the immunogenicity was evaluated by analyzing the expression and distribution of the MHC classes I and II and were compared with untreated organs of equivalent age. The untreated fetal adrenal gland depicted little immunogenicity. However, compared with age-matched untreated control organs, at 2 wk after transplantation, the grafts demonstrated an increased expression of MHC I and II, upregulated throughout the entire adrenal cortex. No signs of MHC-mediated rejection were found. The upregulation of MHC persisted until the eighth week after transplantation. At 3 months after transplantation the expression of MHC I and II returned to the normal pattern of untreated controls. As this study used a purely syngeneic model, the immunologic changes observed could not be induced by a graft vs. host incompatibility, instead they were caused by experimental factors. The expressions of MHC class I and II was increased at 2 wk, but these proteins did not induce a T-cell mediated rejection or cellular infiltration. In conclusion, these findings question the concept of an immunologic privilege of fetal tissue transplants. Instead, experimental factors may modify the tissue's primary immaturity of its MHC. Further investigations must evaluate, whether the increase in MHC expression will have an impact on the rejection of fetal adrenal grafts in allogeneic hosts. PMID- 12756047 TI - Tacrolimus vs. cyclosporine A as primary immunosuppression in pediatric renal transplantation: a NAPRTCS study. AB - Using the North American Renal Transplant Cooperative Study (NAPRTCS) database, we performed a retrospective cohort study of 986 pediatric renal transplant recipients (index transplant 1997-2000) who were treated either with Cyclosporine A (CSA), Mycophenolate Mofetil (MMF) and steroids (n = 766) or tacrolimus (TAC), MMF and steroids (n = 220) to examine potential difference in clinical outcomes between these two groups. In the first year post-transplant, time to first rejection (29.1% vs. 29%, p = 0.840), risk for rejection [Adjusted Relative Risk (aRR) 1.01, 95% Confidence Interval (CI) 0.77, 1.323], graft survival (96.8% vs. 97.9%, p = 0.607) and risk for graft failure (aRR 0.988, 95% CI 0.64, 1.928) were not significantly different in TAC and CSA-treated patients. At 2 yr post transplant, there was also no difference in risk for rejection (aRR 0.918, 95% CI 0.669, 1.259), graft survival (91.4% vs. 95.1%, p = 0.152) and risk for graft failure (aRR 0.702, 95% CI 0.461, 1.762) in the subset of 391 CSA-treated patients and 77 TAC-treated patients on whom 2 yr follow data were available in the database. TAC-treated patients were significantly less likely to require antihypertensive medication at 1 yr [aRR 0.74 (95% CI 0.454, 0.637)] and 2 yr post-transplant [aRR 0.67 (95% CI 0.56, 0.793)]. At 1 yr post-transplant, TAC treated patients enjoyed a higher mean GFR as estimated by the Schwartz formula [89.1 mL/min/1.73 m(2) (SE 2.64) vs. 78.6 mL/min/1.73 m(2) (SE 1.07), p = 0.0003]. In addition, in the subset of patients with 2 yr of follow-up, TAC patients had a higher mean GFR at both 1 yr [98.6 mL/min/1.73 m(2) (SE 3.83) vs. 78.0 mL/min/1.73 m(2) (SE 1.44), p = 0.0003] and 2 yr post-transplant [96.7 mL/min/1.73 m(2) (SE 3.33) vs. 73.2 mL/min/1.73 m(2) (SE 1.48), p < 0.0001]. In summary, TAC and CSA, in combination with MMF and steroids, produce similar rejection rates and graft survival in pediatric renal transplant recipients. However, TAC is associated with improved graft function at 1 and 2 yr post transplant. Further analysis as more patient data are obtained will be necessary to determine if this difference in graft function persists and translates into improved graft survival. PMID- 12756048 TI - Limitations of EBV-PCR monitoring to detect EBV associated post-transplant lymphoproliferative disorder. AB - Post-transplant lymphoproliferative disorder (PTLD) represents a significant threat to the survival of pediatric transplant recipients. Epstein-Barr (EBV) viral load monitoring using polymerase chain reaction (PCR) has been reported to have a variable sensitively with relatively higher specificity as in an indicator of the development of PTLD. We report two cases of pathologically confirmed PTLD in children who failed to develop sustained increases in their EBV-PCR determined viral loads. We suggest that clinicians should be aware of the potential for false-negative results of EBV-PCR in pediatric transplant recipients. PMID- 12756049 TI - Quality of life of children and adolescents after kidney or liver transplantation: child, parents and caregiver's point of view. AB - A cross-sectional study was performed to assess quality of life (QoL) after kidney or liver transplantation during childhood. Self-questionnaires explored children, adolescent and parent QoL. Seventy-five transplant children, 36 transplant adolescents, 67 mothers, 34 fathers and 67 caregivers filled out the questionnaires; they were compared with a reference population. Children reported a rather good QoL, but their extra-family involvement appeared not as satisfactory as that of the reference population children. Adolescents reported a very high QoL when completing the structured format scale; however, their responses to open-ended questions showed qualitative differences compared with those of a reference population: they expressed concern about their body or health, less pleasure than ordinary adolescents to manage by themselves, and a poor relationship with peers. Mothers indicated a deep impact of the child's illness on their own QoL, and the need for psychological support. In conclusion, a rather good QoL is a long-lasting feature of kidney and liver transplantation in children. Such an assessment is mandatory to identify remaining impairments in some selected areas, either in the recipient or family. PMID- 12756050 TI - Pneumatosis intestinalis and diarrhea in a child following renal transplantation. AB - Pneumatosis intestinalis is an uncommon finding beyond the neonatal period, but it has been reported in immunocompromized pediatric patients. The association of pneumatosis intestinalis in children following renal transplantation has to the best of our knowledge been only reported once in children. We describe a 4-year old female who developed intermittent emesis, weight loss, and intermittently loose bloody stools after cadaveric renal transplantation at age 3.5 years. An abdominal x-ray demonstrated extensive pneumatosis in the colon. The infectious work-up was negative. Histologically, she had increased eosinophils throughout the lamina propria in the rectum. A glucose breath test was suggestive of small bowel bacterial overgrowth. She was treated with 10 days of metronidazole with resolution of the diarrhea and occult blood in stools. One month after the treatment she had radiologic resolution of her pneumatosis. Based on this report, pneumatosis intestinalis should be considered in the differential diagnosis of children after organ transplant suffering from diarrhea, abdominal pain, or blood in the stool. PMID- 12756051 TI - Liver and intestinal transplantation in a child with cystic fibrosis: a case report. AB - Cystic fibrosis (CF) is an inherited disorder that presents as a multisystem disease with meconium ileus being the presenting symptom in 20% of patients. Approximately half of these patients present with complicated meconium ileus mandating early surgical intervention, potentially resulting in short gut syndrome. Although liver transplantation in children with CF has been described, this is the first report of a combined liver and small bowel transplant in a recipient with CF. A 7-month-old boy with CF presented with short bowel syndrome following extensive small bowel resection for meconium ileus and progressive cholestatic liver failure from intravenous hyperalimentation. He underwent combined liver and small intestinal transplant. He was discharged home three weeks post-transplant on enteral feeds with supplemental intravenous fluid. He has had routine protocol small bowel allograft biopsies with no documented rejection episodes. He has been treated for minor respiratory infections without major sequelae. Improvements in pulmonary therapy have impacted on the survival in the CF population to the point where the need for multiorgan transplantation will be increased in the future. Extrapolating from the excellent experience of liver transplantation in children with CF, early liver and small intestinal multivisceral transplantation, if indicated, can be performed safely in children with CF. PMID- 12756052 TI - Use of appendix for complete transplant ureteral necrosis. AB - A 3-yr-old boy with posterior urethral valves underwent cadaveric renal transplant. On the ninth day after transplantation the patient developed a urinary leak, with complete ureteral necrosis. There was insufficient length of undamaged ureter to permit ureteroneocystostomy, unavailability of a native ureter to permit ureteroureterostomy, and an inability to mobilize the transplant kidney or bladder sufficiently to permit direct pyelovesicostomy. As the kidney was otherwise functioning perfectly, we decided to create an appendiceal conduit in the hope of salvaging the patient's renal allograft. At present, 7 months post transplant, the child is clinically well with a serum creatinine of 0.7 mg/dL. Complete ureteral necrosis is an infrequent but devastating complication following renal transplantation. We report a novel method that allowed an otherwise normally functioning cadaveric graft to be salvaged. PMID- 12756053 TI - Use of basiliximab in pediatric liver transplantation for Langerhans cell histiocytosis. AB - This report describes a 16-month-old girl with multi-system Langerhans cell histiocytosis (LCH), who developed end-stage liver disease despite intensive chemotherapy. She underwent a liver transplant at 28 months of age while receiving maintenance chemotherapy for bony lesions. In view of previous reports of a high incidence of acute cellular rejection and post-transplant lymphoproliferative disease (PTLD) in children transplanted for LCH, basiliximab was added to the post-transplant immunosuppression regime of tacrolimus and prednisolone. Sixteen months post-transplant, she has had no episodes of acute rejection or PTLD and her LCH has remained in remission. Current literature regarding liver transplantation (LTx) for LCH and the use of basiliximab in pediatric LTx is reviewed. PMID- 12756054 TI - Evaluation of support groups for women with breast cancer: importance of the navigator role. AB - BACKGROUND: At least some forms of breast cancer are increasingly being viewed as a chronic illness, where an emphasis is placed on meeting the various ongoing needs of people living with cancer, their families and other members of their social support networks. This commentary outlines some approaches to the evaluation of cancer-related support groups, with a particular emphasis on those designed to provide long-distance support, via the internet, for women with breast cancer. DISCUSSION: The literature on evaluations of community-based cancer support groups indicates that they offer a number of benefits, and that it is more reasonable to expect an impact of such interventions on psychosocial functioning and/or health-related quality of life than on survival. The literature on both face-to-face and online social support groups suggests that they offer many advantages, although evaluation of the latter delivery mechanism presents some ethical issues that need to be addressed. Many popular online support groups are peer-moderated, rather than professionally-moderated. In an evaluation of online support groups, different models of the role of the "navigator" need to be taken into account. Some conceptual models are outlined for the evaluation of the "navigator role" in meeting the informational, decisional and educational needs of women with breast cancer. The Breast-Cancer Mailing List, an example of an unmoderated internet-based peer-support group, is considered within the context of a Shared or Tacit Model of the navigator role. CONCLUSION: Application of the concept of a "navigator role" to support groups in general, and to unmoderated online ones in particular, has received little or no attention in the research literature. The navigator role should be taken into account in research on this increasingly important aspect of cancer communication. PMID- 12756055 TI - Womens' opinions on antenatal care in developing countries: results of a study in Cuba, Thailand, Saudi Arabia and Argentina. AB - BACKGROUND: The results of a qualitative study carried out in four developing countries (Cuba, Thailand, Saudi Arabia and Argentina) are presented. The study was conducted in the context of a randomised controlled trial to test the benefits of a new antenatal care protocol that reduced the number of visits to the doctor, rationalised the application of technology, and improved the provision of information to women in relation to the traditional protocol applied in each country. METHODS: Through focus groups discussions we were able to assess the concepts and expectations underlying women's evaluation of concepts and experiences of the care received in antenatal care clinics. 164 women participated in 24 focus groups discussion in all countries. RESULTS: Three areas are particularly addressed in this paper: a) concepts about pregnancy and health care, b) experience with health services and health providers, and c) opinions about the modified Antenatal Care (ANC) programme. In all three topics similarities were identified as well as particular opinions related to country specific social and cultural values. In general women have a positive view of the new ANC protocol, particularly regarding the information they receive. However, controversial issues emerged such as the reduction in the number of visits, particularly in Cuba where women are used to have 18 ANC visits in one pregnancy period. CONCLUSION: Recommendations to improve ANC services performance are being proposed. Any country interested in the application of a new ANC protocol should regard the opinion and acceptability of women towards changes. PMID- 12756056 TI - Seasonal aspects of sleep in the Djungarian hamster. AB - BACKGROUND: Changes in photoperiod and ambient temperature trigger seasonal adaptations in the physiology and behaviour of many species, including the Djungarian hamster. Exposure of the hamsters to a short photoperiod and low ambient temperature leads to a reduction of the polyphasic distribution of sleep and waking over the light and dark period. In contrast, a long photoperiod enhances the daily sleep-wake amplitude leading to a decline of slow-wave activity in NREM sleep within the light period. It is unknown whether these changes can be attributed specifically to photoperiod and/or ambient temperature, or whether endogenous components are contributing factors. The influence of endogenous factors was investigated by recording sleep in Djungarian hamsters invariably maintained at a low ambient temperature and fully adapted to a short photoperiod. The second recording was performed when they had returned to summer physiology, despite the maintenance of the 'winter' conditions. RESULTS: Clear winter-summer differences were seen in sleep distribution, while total sleep time was unchanged. A significantly higher light-dark cycle modulation in NREM sleep, REM sleep and waking was observed in hamsters in the summer physiological state compared to those in the winter state. Moreover, only in summer, REM sleep episodes were longer and waking bouts were shorter during the light period compared to the dark period. EEG power in the slow-wave range (0.75-4.0 Hz) in both NREM sleep and REM sleep was higher in animals in the summer physiological state than in those in the 'winter' state. In winter SWA in NREM sleep was evenly distributed over the 24 h, while in summer it decreased during the light period and increased during the dark period. CONCLUSION: Endogenous changes in the organism underlie the differences in sleep-wake redistribution we have observed previously in hamsters recorded in a short and long photoperiod. PMID- 12756057 TI - Identification of regeneration-associated genes after central and peripheral nerve injury in the adult rat. AB - BACKGROUND: It is well known that neurons of the peripheral nervous system have the capacity to regenerate a severed axon leading to functional recovery, whereas neurons of the central nervous system do not regenerate successfully after injury. The underlying molecular programs initiated by axotomized peripheral and central nervous system neurons are not yet fully understood. RESULTS: To gain insight into the molecular mechanisms underlying the process of regeneration in the nervous system, differential display polymerase chain reaction has been used to identify differentially expressed genes following axotomy of peripheral and central nerve fibers. For this purpose, axotomy induced changes of regenerating facial nucleus neurons, and non-regenerating red nucleus and Clarke's nucleus neurons have been analyzed in an intra-animal side-to-side comparison. One hundred and thirty five gene fragments have been isolated, of which 69 correspond to known genes encoding for a number of different functional classes of proteins such as transcription factors, signaling molecules, homeobox-genes, receptors and proteins involved in metabolism. Sixty gene fragments correspond to genomic mouse sequences without known function. In situ-hybridization has been used to confirm differential expression and to analyze the cellular localization of these gene fragments. Twenty one genes (approximately 15%) have been demonstrated to be differentially expressed. CONCLUSIONS: The detailed analysis of differentially expressed genes in different lesion paradigms provides new insights into the molecular mechanisms underlying the process of regeneration and may lead to the identification of genes which play key roles in functional repair of central nervous tissues. PMID- 12756058 TI - The effects of estrogen, its antagonist ICI 182, 780, and interferon-tau on the expression of estrogen receptors and integrin alphaV beta 3 on cycle day 16 in bovine endometrium. AB - We have shown previously that downregulation of intercaruncular stromal integrin alphavbeta3 in bovine endometrium on day 16 of the estrous cycle coincided with the antibody recognition of estrogen receptors (ER) in the luminal epithelium. In pregnancy, these changes were not observed. Our hypothesis was that on day 16 of the estrous cycle, estrogen from the dominant follicle causes a reduction in integrin alphavbeta3 and affects ERalpha in the luminal epithelium. The pregnancy recognition protein, interferon-tau (IFN-tau), may prevent downregulation of integrin alphavbeta3 and suppress ERalpha expression in the luminal epithelium. On days 14 to 16, heifers received uterine infusions of the anti-estrogen ICI 182, 780, estradiol 17beta, IFN-tau or the saline control. On day 16, reproductive tracts were collected for analysis of integrin alphavbeta3 and ERalpha. Estrogen receptor alpha immunoreactivity was largely restricted to the luminal epithelium in control animals. Using anti-ERalpha recognizing the amino terminus, estrogen-treated animals showed reactivity in the stroma, shallow and deep glands and myometrium as is typical of estrus, whereas ICI 182, 870 treated heifers showed little or no reactivity. In contrast, carboxyl terminus-directed antibodies showed a widespread distribution of ERalpha with reactivity detected in the uterine epithelium, stroma and myometrium of both estrogen and ICI 182, 780 treated animals. Heifers treated with IFN-tau had low ERalpha reactivity overall. Control and IFN-tau treated heifers had lower intercaruncular stromal expression of integrin alphavbeta3 in comparison to estrogen and ICI 182, 780 treatments. Overall, the results suggest that on day 16 of the estrous cycle, estrogen effects on integrin alphavbeta3 are indirect and do not directly involve ERalpha in the luminal epithelium. During pregnancy, interferon-tau may block ERalpha in the luminal epithelium but likely does not rescue integrin alphavbeta3 expression. PMID- 12756060 TI - Do English and Chinese EQ-5D versions demonstrate measurement equivalence? An exploratory study. AB - BACKGROUND: Although multiple language versions of health-related quality of life instruments are often used interchangeably in clinical research, the measurement equivalence of these versions (especially using alphabet vs pictogram-based languages) has rarely been assessed. We therefore investigated the measurement equivalence of English and Chinese versions of the EQ-5D, a widely used utility based outcome instrument. METHODS: In a cross-sectional study, either EQ-5D version was administered to consecutive outpatients with rheumatic diseases. Measurement equivalence of EQ-5D item responses and utility and visual analog scale (EQ-VAS) scores between these versions was assessed using multiple regression models (with and without adjusting for potential confounding variables), by comparing the 95% confidence interval (95%CI) of score differences between these versions with pre-defined equivalence margins. An equivalence margin defined a magnitude of score differences (10% and 5% of entire score ranges for item responses and utility/EQ-VAS scores, respectively) which was felt to be clinically unimportant. RESULTS: Sixty-six subjects completed the English and 48 subjects the Chinese EQ-5D. The 95%CI of the score differences between these versions overlapped with but did not fall completely within pre-defined equivalence margins for 4 EQ-5D items, utility and EQ-VAS scores. For example, the 95%CI of the adjusted score difference between these EQ-5D versions was -0.14 to +0.03 points for utility scores and -11.6 to +3.3 points for EQ-VAS scores (equivalence margins of -0.05 to +0.05 and -5.0 to +5.0 respectively). CONCLUSION: These data provide promising evidence for the measurement equivalence of English and Chinese EQ-5D versions. PMID- 12756062 TI - Immunity to Trichuris muris in the laboratory mouse. AB - Of all the laboratory models of intestinal nematode infection, Trichuris muris in the mouse is arguably the most powerful. This is largely due to the fact that the ability to expel this parasite is strain dependent. Thus, most mouse strains readily expel T. muris. However certain mouse strains, and indeed some individuals within particular mouse strains, are unable to mount a protective immune response and harbour long term chronic infections. This unique model thus presents an opportunity to examine the immune events underlying both resistance to infection and persistent infection within the same host species, and in some cases, the same host strain. PMID- 12756059 TI - Benefits of psychosocial oncology care: improved quality of life and medical cost offset. AB - The burden of cancer in the worldwide context continues to grow, with an increasing number of new cases and deaths each year. A significant proportion of cancer patients at all stages of the disease trajectory will suffer social, emotional and psychological distress as a result of cancer diagnosis and treatment. Psychosocial interventions have proven efficacious for helping patients and families confront the many issues that arise during this difficult time. This paper reviews the literature detailing the extent of distress in patients, the staffing needed to treat such levels of distress, and the efficacy of psychosocial treatments for cancer patients. This is followed by a summary of the literature on medical cost offset in mental health, other medical populations, and in cancer patients, which supports the notion that psychosocial interventions are not only effective, but also economical. Conclusions support taking a whole-person approach, as advocated by a growing number of health care professionals, which would not only help to treat the emotional and social aspects of living with cancer, but also provide considerable long-term cost savings to overburdened health-care systems. PMID- 12756063 TI - Chasing the genes that control resistance to gastrointestinal nematodes. AB - The host-protective immune response to infection with gastrointestinal (GI) nematodes involves a range of interacting processes that begin with recognition of the parasite's antigens and culminate in an inflammatory reaction in the intestinal mucosa. Precisely which immune effectors are responsible for the loss of specific worms is still not known although many candidate effectors have been proposed. However, it is now clear that many different genes regulate the response and that differences between hosts (fast or strong versus slow or weak responses) can be explained by allelic variation in crucial genes associated with the gene cascade that accompanies the immune response and/or genes encoding constitutively expressed receptor/signalling molecules. Major histocompatibility complex (MHC) genes have been recognized for some time as decisive in controlling immunity, and evidence that non-MHC genes are equally, if not more important in this respect has also been available for two decades. Nevertheless, whilst the former have been mapped in mice, only two candidate loci have been proposed for non-MHC genes and relatively little is known about their roles. Now, with the availability of microsatellite markers, it is possible to exploit linkage mapping techniques to identify quantitative trait loci (QTL) responsible for resistance to GI nematodes. Four QTL for resistance to Heligmosomoides polygyrus, and additional QTL affecting faecal egg production by the worms and the accompanying immune responses, have been identified. Fine mapping and eventually the identification of the genes (and their alleles) underlying QTL for resistance/susceptibility will permit informed searches for homologues in domestic animals, and human beings, through comparative genomic maps. This information in turn will facilitate targeted breeding to improve resistance in domestic animals and, in human beings, focused application of treatment and control strategies for GI nematodes. PMID- 12756064 TI - Biological variation in Trichinella species and genotypes. AB - At present, the genus Trichinella comprises seven species of which five have encapsulated muscle larvae (T. spiralis, T. nativa, T. britovi, T. nelsoni and T. murrelli) and two do not (T. pseudospiralis and T. papuae) plus three genotypes of non-specific status (T6, T8 and T9). The diagnostic characteristics of these species are based on biological, biochemical and genetic criteria. Of biological significance is variation observed among species and isolates in parameters such as infectivity and immunogenicity. Infectivity of Trichinella species or isolates is determined, among other considerations, by the immune status of the host in response to species- or isolate-specific antigens. Common and particular antigens determine the extent of protective responses against homologous or heterologous challenge. The kinetics of isotype, cytokine and inflammatory responses against T. spiralis infections are isolate-dependent. Trichinella spiralis and T. pseudospiralis induce different dose-dependent T-cell polarizations in the early host response, with T. spiralis initially preferentially promoting Th1-type responses before switching to Th2 and T. pseudospiralis driving Th2-type responses from the outset. PMID- 12756065 TI - Responses of inbred mouse strains to infection with intestinal nematodes. AB - Comparisons were made of the immune and inflammatory responses of four strains of inbred mice to infection with the intestinal nematodes Trichinella spiralis and Nippostrongylus brasiliensis to determine whether genetically determined 'high responsiveness' to infection, seen most clearly in intestinal responses, is independent of the parasite concerned and necessarily correlated with protection. The time course of infection was followed by counting adult worms at intervals after infection. Mucosal mast cells and Paneth cell numbers were determined as indices of the intestinal inflammatory response. Levels of IgG2a and IgG1 antibodies and of the cytokines IFN-gamma and IL-5 released from in vitro stimulated mesenteric node lymphocytes were measured to assess type 1 and type 2 responses. NIH and CBA mice were the most resistant to T. spiralis and N. brasiliensis respectively, resistance in each case being correlated with the most intense intestinal inflammatory responses. C57BL/10 (B10) and B10.BR were the least resistant to T. spiralis, but were as resistant as CBA to N. brasiliensis, despite their intestinal inflammatory responses to both parasites being much lower than the other two strains. Mice infected with T. spiralis made the expected switch from a type 1 (IFN-gamma) to a type 2 (IL-5) response between days 2 and 8, and there were no significant differences in levels of these cytokines between the strains. In contrast, when infected with N. brasiliensis, CBA showed an IFN-gamma response at day 4, all strains switching to IL-5 by day 8 and NIH mice releasing the greatest amount of IL-5. The results indicate that the "high responder" phenotype to intestinal nematode infection is in part determined by host characteristics, but is also determined by the parasite concerned--seen most clearly by the differences between NIH and CBA when infected with T. spiralis and N. brasiliensis. The fact that "low responder" B10 background mice were more resistant to N. brasiliensis than "high responder" NIH implies that each parasite elicits a particular pattern of protective host responses, rather than parasites being differentially susceptible to the same response profile. PMID- 12756066 TI - Host-parasite interactions in rodent nematode infections. AB - In rodents, Trichinella spiralis and Nippostrongylus brasiliensis infect the small intestine and Trichuris muris resides in the colon. The intestinal host response in these animals is characterized by changes in mucosal architecture and inflammation and is associated with worm expulsion. The requirement of T cell mediated host response in worm expulsion has been demonstrated over many years. Subsequent studies have shown that Th2-type, but not Th1-type, responses mediate resistance to the nematodes. Investigations using neutralizing antibodies and genetically manipulated mice have characterized the contribution of individual Th2-type cytokines in not only worm expulsion, but also specific cellular changes that occur in the mucosa, such as alterations in epithelial phenotype and smooth muscle. There is also increasing appreciation of the contribution of non-bone marrow-derived cells in innate and adaptive host responses in these models. PMID- 12756067 TI - The relative involvement of Th1 and Th2 associated immune responses in the expulsion of a primary infection of Heligmosomoides polygyrus in mice of differing response phenotype. AB - T helper cell (Th1 and Th2) associated responses were examined following a primary infection with the gastrointestinal nematode Heligmosomoides polygyrus in five inbred strains of mice with different resistance phenotypes. Levels of (i) mast cell protease, (ii) specific IgE, (iii) nitric oxide and (iv) specific IgG2a, as markers of Th2 and Th1 associated responses, respectively, were determined in sera and intestinal fluids and correlated with worm burdens. The "fast" responder (resistant) strains SWR and SJL produced strong Th2 and Th1 associated responses respectively in a mutually exclusive fashion. The F1 hybrid (SWRxSJL) F1, showed rapid expulsion of the parasite and expressed both intense Th1 and Th2 responses, suggesting synergism between Th1 and Th2 activity in these mice. The results indicate that both Th2 and Th1 responses operate in mice following a primary infection with H. polygyrus and that each Th response may be involved to a greater or lesser degree within certain strains. Resistance to H. polygyrus was found to correlate only to the intensity of either the gut associated mastocytosis or nitric oxide production in these strains but not to either specific IgE or IgG2a titres. Chronic infections in the "slow" response phenotype mouse strains CBA and C57BL/10, were associated with both poor Th2 and poor Th1-associated responses attributed to a general parasite-mediated immunosuppression of the host immune response to infection. PMID- 12756068 TI - Causality or coincidence: may the slow disappearance of helminths be responsible for the imbalances in immune control mechanisms? AB - Intestinal infection continues to be a problem worldwide and helminths, which currently infect billions of individuals, are primary culprits. The major burden of disease falls on the populations of developing countries, given that over the last four to five decades helminth infections are disappearing in industrialized societies. In developing countries, a major source of immunomodulatory signals in post-natal life are parasites, particularly helminths, which, unlike most bacteria and viruses, selectively stimulate Th2 function. Helminths and their eggs are probably the most potent stimulators of mucosal Th2 responses. Responses elicited by worms can modulate immune reactions to other parasites, bacterial, viral infections and several unrelated diseases. Bacterial and protozoal infections may also protect against atopy and asthma, through the induction of the Th1 regulatory responses. Today, people in developed countries often live in ultra-hygienic environments, avoiding exposure to viruses, bacteria, ectoparasites and endoparasites, particularly helminths. Perhaps failure to acquire worms and experience mucosal Th2 conditioning predisposes to unrelated diseases. In contrast to this hypothesis it has also been suggested that Th2 responses can make the host more susceptible to other important diseases and to contribute to the spread of them. PMID- 12756069 TI - Mucosal mast cells and nematode infection: strain-specific differences in mast cell precursor frequency revisited. AB - Mucosal mast cells (MMC) play an important role in the immune response against selected species of intestinal nematode. The kinetics with which different strains of inbred mice resolve infection with Trichinella spiralis correlates with their ability to mount MMC responses in the intestinal mucosa. Homologues of MMC that express and constitutively secrete abundant amounts of the granule chymase, mouse mast cell protease-1 (mMCP-1), can be generated in vitro from bone marrow cultures supplemented with interleukins-3 and -9, stem cell factor and transforming growth factor-beta1. Using the enhanced growth characteristics of these MMC homologues, a novel limiting dilution assay for mast cell precursor (MCp) frequency has been developed. The assay is highly specific, in that cultures containing mast cells are identified with mMCP-1 specific antibody, and almost three-fold more sensitive than previously published systems. MCp frequencies were compared in BALB/c and C57/BL10 strains of mice that, respectively, respond rapidly and slowly to infection with T. spiralis. MCp frequency (1/378 bone marrow cells) was significantly greater in BALB/c than C57/BL10 mice (frequency: 1/751). Similarly the rate of growth of MMC homologues and the production of mMCP-1 was significantly greater in BALB/c than in C57/BL10 bone marrow cultures. PMID- 12756070 TI - The occurrence and infection dynamics of Anisakis larvae in the black-scabbard fish, Aphanopus carbo, chub mackerel, Scomber japonicus, and oceanic horse mackerel, Trachurus picturatus from Madeira, Portugal. AB - Larval stages of Anisakis spp. (Nematoda: Anisakidae) were found encapsulated or free in the viscera and abdominal cavity of the black-scabbard fish, Aphanopus carbo, chub mackerel, Scomber japonicus and oceanic horse mackerel, Trachurus picturatus in Madeiran waters. The prevalence of infection reached 97.2% for A. carbo, 69.5% for S. japonicus and 62.5% for T. picturatus. Considerable differences in parasite intensities between A. carbo and both S. japonicus and T. picturatus were found, with mean intensities up to 69.6 in A. carbo, while in the other two fish hosts the intensity reached only a maximum of 2.6. These differences were probably due to different feeding behaviours of the hosts. Intensities of Anisakis sp. in A. carbo were high irrespective of sex and season. No relationship between host length and prevalence of infection was observed for A. carbo, while for S. japonicus a weak positive significant relationship was found. PMID- 12756071 TI - Serological responses to Ascaris suum adult worm antigens in Iberian finisher pigs. AB - Adult Ascaris suum were dissected to obtain different worm components (body wall, body fluid, ovaries, uterus and oesophagus) which were used as antigens when testing 95 sera of naturally A. suum-infected Iberian pigs by enzyme-linked immunosorbent assay (ELISA) and Western blot (WB). Pigs with patent Ascaris infections had significantly lower ELISA optical density values than pigs without adult worms when using the body fluid and the body wall as antigens. A poor negative correlation was found between adult intestinal worm burden or eggs in faeces and specific antibody responses, measured by ELISA and WB using all antigens. By WB, the recognition of specific bands was variable, but three groups of bands with molecular weights of 97 kDa, 54-58 kDa and 42-44 kDa were generally recognized by sera from naturally infected pigs as well as from hyperimmunized pigs when using the five antigen extracts. The ELISA and WB techniques may be used for immunodiagnosis, using somatic adult worm antigens, to declare young pigs to be Ascaris-free but cannot be used for individual Ascaris-diagnosis in adult Iberian pigs. PMID- 12756072 TI - Potential interactions between metazoan parasites of the Mayan catfish Ariopsis assimilis and chemical pollution in Chetumal Bay, Mexico. AB - The effect of pollutants on the intensity of infection of metazoan parasites in the Mayan catfish, Ariopsis assimilis was investigated. Data were collected on pollutants and metazoan parasites from 76 catfish from five localities in Chetumal Bay in October, 1996. Nineteen pollutants (pesticides, polychlorinated biphenyls (PCBs) and polycyclic aromatic hydrocarbons (PAHs)) were found in the catfish livers. Heavy metal content was not determined. Nineteen metazoan parasite species were recovered. After controlling for fish length and sampling station, there was a significant negative linear relationship between the intensity of the larval digenean Mesostephanus appendiculatoides and 1,1,1, trichloro-2,2-bis (4-chlorophenyl) ethane (DDT) concentrations. This negative relationship may be explained either by the effect of the pesticide on the mortality of (i) free-living larval forms, (ii) metacercariae in the fish, (iii) infected fish or (iv) intermediate host snails. There were significant differences between fish parasitized and not parasitized with M. appendiculatoides with respect to their DDT concentrations. There were also significant differences between the variances of the mean Clark's coefficient of condition values between catfish parasitized and not parasitized by M. appendiculatoides, with the variance of non-parasitized catfish being significantly larger. The results provided statistical evidence that DDT has a detrimental effect on M. appendiculatoides infection intensity. Furthermore, the significantly larger variance value of Clark's coefficient for non-parasitized fish suggested that DDT affects both the parasite and general host condition. PMID- 12756074 TI - Neoadjuvant chemotherapy for operable breast cancer: is this the future? AB - The idea of using preoperative or neoadjuvant chemotherapy in patients with operable breast cancer originated from experimental and clinical observations as well as theoretical hypotheses on tumor cell growth and dissemination. Initially, nonrandomized studies demonstrated considerable rates of clinical tumor response, low rates of pathologic complete response (pCR), and increased rates of breast conserving procedures. However, nonrandomized studies could not address the relative efficacy of neoadjuvant versus adjuvant chemotherapy on disease-free and overall survival. Similarly, earlier randomized trials were not designed as straightforward comparisons of neoadjuvant versus adjuvant chemotherapy and therefore could not adequately address the relative efficacy of neoadjuvant versus adjuvant chemotherapy on outcome. These answers were eventually provided by larger randomized trials that directly compared neoadjuvant with adjuvant chemotherapy, which are reviewed in more detail in this article. Potential advantages and disadvantages of the neoadjuvant approach and surgical considerations in the breast and axilla after neoadjuvant chemotherapy are also discussed. Finally, several recently reported trials of neoadjuvant therapy incorporating newer agents such as taxanes in sequence with anthracycline containing regimens have shown further increases in pCR rates. Although outcome data are not available yet from these studies, it is hoped that the observed increase in pCR rates will be associated with improved outcome. If the previously observed significant correlation between the achievement of pCR and improved outcome continues to be demonstrated with these newer regimens, it will substantially strengthen the rationale for using neoadjuvant rather that adjuvant chemotherapy in the clinical setting as well as in future research studies. PMID- 12756075 TI - The evolving role of capecitabine in breast cancer. AB - Breast cancer is the most common cancer and the second greatest cause of cancer related death among women in the United States. Capecitabine is a selectively tumor-activated fluoropyrimidine carbamate that is converted to 5-fluorouracil by the sequential activity of these enzymes, the final of which is thymidine phosphorylase, which is overexpressed in many human cancers. Capecitabine as a single agent and in combination with other drugs is efficacious in previously treated and untreated metastatic breast cancer (MBC). The integration of capecitabine, either as a single agent or in combination with docetaxel, into adjuvant breast cancer therapy is justified due to its high antitumor activity in previously treated and untreated MBC, its tolerability, lack of cross-resistance with the anthracyclines and taxanes, and because combined docetaxel/capecitabine improves the overall survival of patients with MBC. Capecitabine is being evaluated as preoperative therapy in patients with operable breast cancer, as adjuvant therapy in patients with high-risk node-negative or node-positive disease, and as oral single-agent therapy in women > or = 65 years of age. This article is an overview of published studies of capecitabine in MBC and the studies that are planned or have been proposed to evaluate capecitabine as adjuvant therapy for breast cancer. PMID- 12756076 TI - Epirubicin versus doxorubicin: which is the anthracycline of choice for the treatment of breast cancer? AB - Breast cancer is the most common malignancy in women in the United States. The addition of anthracyclines to adjuvant therapy regimens has resulted in improvement in overall survival of patients. The 2 most commonly used anthracyclines are doxorubicin and epirubicin. Comparison studies in the metastatic setting have shown that, at similar doses, these 2 anthracyclines provide similar response rates. However, their toxicity profiles differ somewhat. The major side effects of anthracyclines are cardiotoxicity and myelosuppression. The equimolar dose ratios of doxorubicin to epirubicin for myelosuppression and cardiotoxicity are 1:1.2 and 1:1.7-2.0, respectively. There have been many studies comparing different schedules and doses of anthracyclines in the adjuvant setting. However, direct comparisons between doxorubicin and epirubicin in early stage breast cancer have not been performed to date. In this article, we are attempting to provide an overview of current use of doxorubicin and epirubicin in breast cancer PMID- 12756077 TI - Human epidermal growth factor receptor-2 and hormonal therapies: clinical implications. AB - Estrogen-targeted therapies such as administration of tamoxifen or aromatase inhibitors are among the most important treatment strategies in the modern management of breast cancer. Despite initial responses in the metastatic setting and prolonged disease-free intervals in the adjuvant setting, many patients subsequently become resistant to these agents. Human epidermal growth factor receptor-2 (HER2) is a transmembrane glycoprotein receptor that is overexpressed in 13%-30% of human breast cancers. There are experimental data suggesting an important role for HER2 in de novo and acquired resistance to endocrine therapies. These experimental data are discussed in this article, as are clinical data addressing the role of HER2 in resistance to endocrine therapy in the adjuvant, neoadjuvant, and metastatic settings. Responses and benefit from tamoxifen appear to be impaired in patients in whom HER2 is overexpressed. In contrast, early data from the neoadjuvant setting suggest that responses to aromatase inhibitors may be maintained in patients with HER2 overexpression. PMID- 12756078 TI - Anastrozole as adjuvant therapy for early-stage breast cancer: implications of the ATAC trial. AB - Tamoxifen has been the gold standard adjuvant therapeutic agent for postmenopausal women with hormone-sensitive breast cancer for > 25 years. Although it continues to play an important role in treating premenopausal women, tamoxifen's association with some serious safety and tolerability issues, including increased incidence of endometrial cancer and thromboembolic events, may be cause to limit its use in postmenopausal women. Anastrozole was the first drug to show improved efficacy and safety compared with standard therapies for first- and second-line therapy of hormone-sensitive advanced breast cancer in postmenopausal women. This article provides a review of the results of the first major analysis of the Arimidex, Tamoxifen, Alone or in Combination (ATAC) early stage breast cancer trial, initiated in 1996, and discusses the implications for the use of anastrozole in the adjuvant setting. This randomized, double-blind, multicenter trial compared tamoxifen (20 mg once daily) with anastrozole (1 mg) alone and in combination with tamoxifen, as adjuvant endocrine treatment for postmenopausal patients with operable, invasive, early-stage breast cancer. The results of the ATAC trial show anastrozole to be more effective and better tolerated than tamoxifen in this group of patients, and an updated follow-up suggests the therapeutic index for anastrozole will continue to remain superior to that of tamoxifen. Anastrozole is now emerging as a new standard for the adjuvant treatment of postmenopausal women with hormone-sensitive early-stage breast cancer. New adjuvant trials are currently using anastrozole in the control arm. PMID- 12756079 TI - Cytokeratin staining and other sentinel node controversies. AB - Sentinel lymph node (SLN) biopsy for breast cancer staging has been widely accepted because it is more sensitive and less morbid than axillary dissection. Sentinel nodes can be thoroughly scrutinized using a variety of techniques increasing the detection of micrometastases; however, the clinical relevance of micrometastases has been challenged. The available data suggest that the prognostic significance of axillary metastases is related to the size of the metastases, and the best data suggest that outcome for patients with metastases < 0.2 mm is similar to patients with node-negative disease. This would argue against the use of ultrasensitive tests such as reverse transcriptase polymerase chain reaction. Immunohistochemistry upstages 2%-20% of hematoxylin and eosin negative sentinel nodes, and additional nodal metastases are identified in approximately 10% of completion axillary dissections prompted by an immunohistochemistry (IHC)-positive sentinel node. This would appear to be a good reason to perform IHC and act on the results. Because micrometastases can be artifactual, SLN biopsy in ductal carcinoma in situ can lead to harmful overtreatment and is best performed in the context of clinical trials. Lymphoscintigraphy has allowed the detection of alternate drainage patterns to internal mammary, infraclavicular, and supraclavicular lymph nodes. Although patients are occasionally identified who have metastases to these basins but not the axilla, this information will not impact the decision for chemotherapy in most cases. Internal mammary SLN biopsy may have value in patients with tumors < 1 cm, but requires additional evaluation in clinical trials. PMID- 12756080 TI - Positron emission tomography in diagnosis and management of invasive breast cancer: current status and future perspectives. AB - [18F]fluorodeoxyglucose positron emission tomography (FDG-PET) is a metabolic imaging modality that has increasing applications in oncology, neurology, and cardiology. Among the oncology applications, breast cancer is one of the most extensively studied diseases. FDG-PET has been performed for diagnosis, staging, and restaging of invasive breast cancer and for monitoring responsiveness to therapies. At the present time, the results of FDG-PET in detection of primary breast cancer and axillary staging are mixed and inconclusive. However, results demonstrating the superiority of FDG-PET over anatomic imaging modalities in detection of distant metastasis, recurrence, and monitoring therapies are relatively well documented. These applications have been accepted by medical professionals and the public, as evidenced by a recent decision by the Centers for Medicare and Medicaid Services (formerly Health Care Financing Agency) to provide coverage for the procedure. Future trends in this exciting area include development of novel breast cancer-specific PET radiopharmaceuticals and use of dedicated breast PET technologies for scans of breast/axillary lesions. PET/computed tomography technology, which combines anatomic and molecular/biochemical information, is also rapidly proliferating and should help to further improve the management of patients with breast cancer. The role of FDG PET in breast cancer is increasing and evolving, and this metabolic imaging modality, in conjunction with newer tracers and other anatomic imaging methods, should improve diagnosis and management of patients with breast cancer PMID- 12756081 TI - Quality of life after treatment for prostate cancer. AB - Quality of life is a major concern of patients when they are choosing treatment for prostate cancer. Health-related quality of life is a patient-centered variable from the field of health services research that can be measured in a valid and reliable manner. Using standardized questionnaires specifically developed to capture health-related quality of life data in men with prostate cancer, the effect of treatments on patients' quality of life can be studied. Patients with localized disease who are undergoing radical prostatectomy tend to have more sexual and urinary dysfunction than men undergoing external beam radiation therapy, although both groups have more impairment in these areas than age-matched controls. Men undergoing external beam radiation therapy have worse bowel function and more urinary distress from irritative voiding symptoms than men undergoing radical prostatectomy or age-matched controls. Recent studies of men undergoing interstitial brachytherapy indicate that these patients have less urinary leakage than those who undergo radical prostatectomy, but experience considerably more irritating voiding symptoms, which often profoundly affect their quality of life. Better information regarding the potential impact of prostate cancer treatment on quality of life will improve medical decision making. PMID- 12756082 TI - Radical prostatectomy for the patient with locally advanced prostate cancer. AB - The proper management of patients with locally advanced adenocarcinoma of the prostate has been contentious and too frequently based on antiquated misconceptions. Non-extirpative treatments, even when combined with neoadjuvant hormonal therapy, are inferior to the surgical removal of the prostate for controlling local progression and distant dissemination of the cancer. Radical prostatectomy combined with early adjunctive hormonal therapy for patients with nodal metastasis is superior to all other forms of therapy and should be considered the standard of care. This approach provides survival rates comparable with patients with clinically organ-confined prostate cancer. PMID- 12756083 TI - Radiotherapy for t3 prostate cancer. AB - Throughout the past decade, significant improvements in optimizing the management of T3 prostate cancer have been made. Phase III randomized studies have demonstrated the superiority of combined hormone therapy and radiation therapy over radiation therapy alone. Radiotherapy dose escalation using conformal techniques has improved local control and lowered toxicity. These advances have changed the approach to treatment of T3 prostate cancer from that directed at palliation of symptoms to earlier interventions directed at prolongation of survival. The contemporary role of radiation therapy in clinical T3 prostate cancer in the context of other treatment options is reviewed in this article. PMID- 12756084 TI - Salvage radical prostatectomy for recurrence of prostate cancer after radiation therapy. AB - The goal of primary radiation therapy in the treatment of prostate cancer is to eradicate all of the local tumor. Although patients with relapsing disease after radiation therapy differ in their risk of death from prostate cancer, many will develop local progression, metastasis, and death. The recognition that local recurrence after radiation therapy portends a poor prognosis has led to the development of improved methods for early detection of recurrence and the development of alternative treatment strategies for radioresistant cancers. This article reviews knowledge regarding radiation failure and the role of salvage radical prostatectomy for men with local recurrence after radiation therapy for prostate cancer. PMID- 12756085 TI - Alternative therapies for localized prostate cancer. AB - Prostate cancer is the leading malignancy in men; an increase in detected localized prostate cancers is expected in the years to come. Radical prostatectomy, although effective, is associated with a considerable morbidity. The aim of minimal invasive alternative treatment options should be equal efficacy, but a decrease in side effects. Cryosurgical ablation of the prostate, brachytherapy, high-intensity focused ultrasound, and radiofrequency interstitial tumor ablation were evaluated after a literature review from a MEDLINE search (1966-2002). When compared with treatments in the 1960s and 1970s, increased safety is observed in all of the alternative treatments available today. Sophisticated technology, including the latest ultrasonography devices for exact planning and monitoring of treatment, contributes largely to this safety. Five year results of cryosurgical ablation of the prostate show a prostate-specific antigen lower than 1 ng/mL in 60% of the cases; in the third generation, there are no long-term data available on cryosurgical ablation of the prostate. Recent outcome data of brachytherapy come close to results of radical prostatectomy series. Brachytherapy is the only true alternative at this point in time. High intensity focused ultrasound and radiofrequency interstitial tumor ablation are promising new technologies that have proven to be able to induce extensive necrosis; however, follow-up is too short to determine their definite places in the treatment of prostate cancer. PMID- 12756086 TI - Photodynamic therapy: a new approach to prostate cancer. AB - Photodynamic therapy (PDT) is based on the concept that light irradiation can change an inert substance into an active one. In urology, hematoporphyrin derivative (HpD) and Photofrin (Axcan Scandipharm Inc., Birmingham, AL) are used most commonly as photosensitizing agents predominantly for the treatment of transitional cell carcinoma of the bladder. To investigate the basics for PDT of prostate cancer, several studies were performed on the optical characteristics of prostate tissue and prostate carcinoma tissue in vitro and in vivo and on the penetration depths of different laser wavelengths. Initial experimental studies to treat prostate cancer with PDT using HpD were done on Dunning tumors in rats. Combined with interstitial applicators, photodynamic therapy seems to have a great potential in the treatment of prostate carcinoma. However, it is an experimental treatment and even a preliminary evaluation will be possible only after the conclusion of clinical studies with the corresponding long-term results. PMID- 12756088 TI - Brachytherapy: update and results. AB - Prostate cancer is the most common noncutaneous malignancy and the second most common cause of cancer mortality in American men. Treatment options for these patients include radical prostatectomy, external beam radiation therapy, hormonal therapy, and prostate brachytherapy. Patients with clinically and radiographically localized disease, especially young patients with few comorbid illnesses, are good candidates for prostate brachytherapy. Prostate brachytherapy has gained widespread acceptance throughout the past two decades and data from several large series of patients are now available. This article describes current techniques, treatment issues, and clinical results of permanent seed implants. PMID- 12756089 TI - Long-term results after external beam radiation therapy for T1-T2 localized prostate cancer. AB - The incidence of organ-confined and early-stage prostate cancer has increased. The external beam radiation therapy has proven to be a good therapeutic option in terms of biochemical survival and overall survival. It has been modified throughout the years; consequently, the available data on the long-term efficacy of external beam radiation therapy are difficult to compare with the commonly used improved radiation strategies. Intensity-modulated conformal radiotherapy and three-dimensional conformal radiotherapy result in better tumor control at a lower complication rate. External beam radiotherapy seems to be favored in intermediate- and high-risk groups for relapse of prostate cancer and radical prostatectomy is favored in the low-risk group. However, they score similarly in terms of general health-related quality of life after treatment. PMID- 12756087 TI - Chemotherapy of prostate cancer: present and future. AB - The role of chemotherapy in prostate cancer continues to evolve. In men with symptomatic androgen-independent prostate cancer, significant reduction in pain and analgesic requirements are achievable with mitoxantrone and glucocorticoid combinations compared with glucocorticoids alone. However, survival rates are not improved. Taxane-based combinations with estramustine phosphate or other new agents show promise. Prostate-specific antigen response rates with these combinations appear to be 1.5 to 2 times more frequent than with mitoxantrone based combinations. Randomized trials of taxane versus mitoxantrone-based therapies are underway. New agents and applications of current agents in adjuvant settings should be explored if survival in men with prostate cancer is to be improved. PMID- 12756090 TI - The status of high-intensity focused ultrasound in the treatment of localized prostate cancer and the impact of a combined resection. AB - To decrease side effects observed after high-intensity focused ultrasound (HIFU) treatment for localized prostate cancer and to re-establish normal micturition in a patient population that often presents with concomitant prostate enlargement, the impact of a combined transurethral resection of the prostate (TURP) and HIFU has been evaluated. TURP and HIFU treatments were performed under the same spinal anesthesia. For the HIFU treatments, the Ablatherm device (EDAP SA, Lyon, France) was used. Selection criteria for HIFU treatment were localized prostate cancer, no previous treatment for prostate cancer, and prostate-specific antigen (PSA) pound 15 ng/mL at diagnosis. All patients meeting these criteria were considered for treatment and analysis. PSA nadir and stability, histology, International Prostate Specific Score (IPSS) and IPSS-quality of life, and morbidity were assessed during follow-up; 271 patients were selected: 96 in the HIFU group and 175 in the TURP plus HIFU group. A statistically significant impact was observed on catheter time (40.0 days versus 7.0 in median), incontinence (15.4% versus 6.9%), urinary infection (47.9% versus 11.4%), and the evolution of the post treatment IPSS (8.91 versus 3.37 in average) in favor of the TURP plus HIFU group. No significant changes were observed regarding efficacy during short-term follow-up when considering a 25% retreatment rate in the HIFU group versus a 4% retreatment rate in the TURP plus HIFU group. The combination of a TURP and HIFU treatment reduces the treatment-related morbidity significantly. The patient management after a combined TURP and HIFU treatment is comparable with the management after a single TURP. PMID- 12756091 TI - PC-SPES for treatment of prostate cancer: herbal medicine. AB - The number of patients who seek treatment with complementary and alternative medicine has increased during the past decade. The trend is primarily driven by consumers who start to change their views toward conventional pharmaceutical approaches that are offered to them. Among all complementary and alternative therapies used in the management of prostate cancer, Prostate Cancer-SPES (PC SPES) has attracted much national attention because of its potency, controversy, and recall by the US Food and Drug Administration. PC-SPES contains extracts from a mixture of eight common herbs that have been used for thousands of years. This article is devoted to reviewing the basic and clinical data of using PC-SPES in prostate cancer therapy. It also explores the difference in philosophies between Western medicine and herbal medicine and explains the inherent difficulties in evaluating herbal medicine. The article concludes that PC-SPES cannot be evaluated by the same standards established to test synthetic pharmaceutical compounds. Thus, new standards need to be developed for the evaluation of herbal medicine. PMID- 12756092 TI - Successful treatment of vitiligo with 0.1% tacrolimus ointment. PMID- 12756093 TI - Maimonides and midcareer. PMID- 12756094 TI - A double-blind randomized trial of 0.1% tacrolimus vs 0.05% clobetasol for the treatment of childhood vitiligo. AB - OBJECTIVE: To assess the safety and efficacy of topical 0.1% tacrolimus vs 0.05% clobetasol propionate. DESIGN: Randomized double-blind trial. SETTING: Department of Dermatology, Hospital Central Dr Ignacio Morones Prieto, San Luis Potosi, Mexico. PARTICIPANTS: From 20 children with vitiligo, 2 symmetrical lesions of about the same size and evolution time were selected. They were devoid of any topical or systemic therapy for 2 months prior to inclusion. Interventions Treatment with topical tacrolimus and clobetasol for a 2-month period. MAIN OUTCOMES MEASURES: The grade of repigmentation was evaluated by color slides at baseline and again at every 2-week visit. The slides were analyzed by 2 clinicians unrelated to the study and by a morphometric digitalized computer program. Characteristics of pigment, time of response, symptoms, telangiectasias, and atrophy were evaluated every 2 weeks. RESULTS: Eighteen (90%) of the 20 patients experienced some repigmentation. The mean percentage of repigmentation was 49.3% for clobetasol and 41.3% for tacrolimus. Lesions in 3 patients using clobetasol presented atrophy, and 2 lesions incurred telangiectasias; tacrolimus caused a burning sensation in 2 lesions. CONCLUSIONS: Tacrolimus proved almost as effective as clobetasol propionate to restore skin color in lesions of vitiligo in children. Because it does not produce atrophy or other adverse effects, tacrolimus may be very useful for younger patients and for sensitive areas of the skin such as eyelids, and it should be considered in other skin disorders currently treated with topical steroids for prolonged periods. PMID- 12756095 TI - Quality of abstracts in 3 clinical dermatology journals. AB - BACKGROUND: Structured abstracts have been widely adopted in medical journals, with little demonstration of their superiority over unstructured abstracts. OBJECTIVES: To compare abstract quality among 3 clinical dermatology journals and to compare the quality of structured and unstructured abstracts within those journals. DESIGN AND DATA SOURCES: Abstracts of a random sample of clinical studies (case reports, case series, and reviews excluded) published in 2000 in the Archives of Dermatology, The British Journal of Dermatology, and the Journal of the American Academy of Dermatology were evaluated. Each abstract was rated by 2 independent investigators, using a 30-item quality scale divided into 8 categories (objective, design, setting, subjects, intervention, measurement of variables, results, and conclusions). Items applicable to the study and present in the main text of the article were rated as being present or absent from the abstract. A global quality score (range, 0-1) for each abstract was established by calculating the proportion of criteria among the eligible criteria that was rated as being present. A score was also calculated for each category. Interrater agreement was assessed with a kappa statistic. Mean +/- SD scores were compared among journals and between formats (structured vs unstructured) using analysis of variance. MAIN OUTCOME MEASURES: Mean quality scores of abstracts by journal and by format. RESULTS: Interrater agreement was good (kappa = 0.71). Mean +/- SD quality scores of abstracts were significantly different among journals (Archives of Dermatology, 0.78 +/- 0.07; The British Journal of Dermatology, 0.67 +/- 0.17; and Journal of the American Academy of Dermatology, 0.64 +/- 0.15; P =.045) and between formats (structured, 0.71 +/- 0.11; and unstructured, 0.56 +/- 0.18; P =.002). The setting category had the lowest scores. CONCLUSIONS: The quality of abstracts differed across the 3 tested journals. Unstructured abstracts were demonstrated to be of lower quality compared with structured abstracts and may account for the differences in quality scores among the journals. The structured format should be more widely adopted in dermatology journals. PMID- 12756096 TI - Histopathologic recognition of involved margins of lentigo maligna excised by staged excision: an interobserver comparison study. AB - OBJECTIVES: To assess interobserver and intraobserver concordance for identifying positive and negative margins in staged excisions of lentigo maligna and lentigo maligna melanoma and to determine if control biopsy specimens are useful to improve concordance. DESIGN: Retrospective, randomized interobserver and intraobserver comparison study of archived pathologic specimens. The study was conducted in 3 phases, and slides were evaluated blindly and independently by 5 pathologists: in phase 1, all slides were randomized and diagnosed as positive or negative. In phase 2, every third slide was evaluated again and diagnosed as positive or negative. In phase 3, slides were organized into cases, allowing evaluation of each margin in the context of the positive control (tumor from the center of the lesion) and negative control (control biopsy specimen), if available. SETTING: University referral center. STUDY MATERIAL: A total of 301 glass microscopic slides from 27 patients who underwent staged excision for lentigo maligna or lentigo maligna melanoma from March 1997 to April 2001. MAIN OUTCOME MEASURES: Interobserver and intraobserver concordance between original diagnoses and study diagnoses rendered on all slides by 5 pathologists. RESULTS: Phase 1 and 3 agreement was moderate (kappa range, 0.4-0.5). Phase 2 (intraobserver) agreement was moderate to good for all pathologists (kappa range, 0.6-0.9). Subset analysis revealed a statistically significant increase in agreement with the use of a control strip biopsy specimen for difficult slides. CONCLUSIONS: Interobserver concordance for margin analysis in lentigo maligna and lentigo maligna melanoma is moderate, and intraobserver concordance is moderate to good. A control strip biopsy specimen may improve concordance in some cases. PMID- 12756097 TI - Dermatologist detection and skin self-examination are associated with thinner melanomas: results from a survey of the Italian Multidisciplinary Group on Melanoma. AB - OBJECTIVE: To investigate patterns of detection and variables associated with early diagnosis of melanoma in a population at intermediate melanoma risk. DESIGN: Survey. SETTING: Hospital and university centers belonging to the Italian Multidisciplinary Group on Melanoma. PATIENTS: Eight hundred sixteen patients who were consecutively diagnosed as having melanoma and treated at 11 participating centers. MAIN OUTCOME MEASURE: Relationship between patterns of detection and patient's and physician's delay with melanoma thickness, assessed by multivariate analysis. RESULTS: A statistically significant association with early diagnosis was found for female sex (odds ratio [OR] for a lesion >1 mm in thickness, 0.70; 95% confidence interval [CI], 0.50-0.97), higher educational level (OR, 0.44; 95% CI, 0.24-0.79), residence in northern and central Italy (compared with southern Italy) (OR, 0.44; 95% CI, 0.30-0.65 and OR, 0.24; 95% CI, 0.15-0.37, respectively), and the habit of performing a skin self-examination (OR, 0.65; 95% CI, 0.45-0.93). When adjusted for all the previously mentioned variables, only melanoma detection made by a dermatologist, maybe incidentally, was associated with a statistically significant additional effect on early diagnosis (OR, 0.45; 95% CI, 0.28-0.73). No significant effect of anatomical site (trunk compared with other sites: OR, 0.83; 95% CI, 0.59-1.17), presence of atypical nevi (OR, 0.78; 95% CI, 0.52-1.17), and patient's delay (>3 months compared with < or =3 months: OR, 1.12; 95% CI, 0.78-1.60) was found. CONCLUSION: Future melanoma early diagnosis strategies should adequately stress the role of skin self-examination among the adult population, and should recommend that dermatologists perform a total skin examination to identify suspect lesions (such an examination should also be performed during consultations for other reasons). PMID- 12756098 TI - Sentinel lymph node biopsy in patients with thin melanoma. AB - OBJECTIVE: To define the percentage of positive sentinel lymph node biopsies and identify risk factors for the presence of lymph node disease in patients with melanomas less than or equal to 1 mm in depth. DESIGN: Retrospective chart review. SETTING: Washington University School of Medicine and Barnes-Jewish Hospital, St Louis, Mo, a melanoma referral center with outpatient surgical care. PATIENTS: Forty-six patients with melanomas less than or equal to 1 mm in depth undergoing sentinel lymph node biopsy at our institution between 1996 and 2002. RESULTS: The procedure was well tolerated and there were no reported complications. Of the 46 patients, 3 (7%) (95% exact confidence interval, 1.3% 17.8%) were found to have positive sentinel lymph nodes or micrometastatic disease. The finding of a positive sentinel lymph node was associated with a Clark level of III or more (P< or =.07). CONCLUSIONS: Conclusions from this study are limited by the small sample size. The results of our study suggest that sentinel lymph node biopsy of patients with melanomas less than or equal to 1 mm in depth may be indicated when the Clark level is III or more. PMID- 12756099 TI - Phase 1/2 pilot study of methotrexate-laurocapram topical gel for the treatment of patients with early-stage mycosis fungoides. AB - OBJECTIVES: To assess the safety and tolerability of a topical gel formulation combining methotrexate and laurocapram and to obtain preliminary information on the therapeutic potential of methotrexate-laurocapram in patients with early stage mycosis fungoides (stage IA or IB). DESIGN: An open-label, phase 1/2 pilot study. SETTING: Two academic referral centers. PATIENTS: Ten patients 18 years or older with histologically confirmed stage IA or IB mycosis fungoides. Intervention The gel formulation of methotrexate-laurocapram was applied to the total body surface, excluding genital, perianal areas, nipples, face, and skin under the breasts, on an every-other-day basis for 24 consecutive weeks. MAIN OUTCOME MEASURES: The safety of methotrexate-laurocapram was assessed in this study by reviewing adverse events and laboratory data. Efficacy outcomes included changes in lesion condition and severity assessments, reduction in area of sample lesions, and the investigator's global evaluation. RESULTS: Adverse events consisted of skin reactions of mild severity. No clinically significant laboratory abnormalities were observed. Based on the investigator's global evaluation at the end of the treatment phase (week 24), 7 (78%) of 9 patients demonstrated a slight-to-moderate response to treatment with methotrexate laurocapram. Statistical significance (P =.049) was reached for induration and pruritus, a trend (P =.10) was observed for erythema, and no change was found for scaling (P =.37). CONCLUSIONS: These findings indicate that the topical administration of methotrexate-laurocapram is safe and in general well tolerated. This treatment may represent a new therapeutic potential for patients with mycosis fungoides. PMID- 12756100 TI - Rapidly growing mycobacterial infections after pedicures. AB - BACKGROUND: Rapidly growing mycobacteria (RGM) can cause a variety of cutaneous and systemic diseases. The causative organisms are typically Mycobacterium fortuitum or Mycobacterium chelonae (also known as Mycobacterium abscessus). Primary cutaneous lesions may develop after a variable latent period, from weeks to several months, and usually result from direct inoculation after trauma, from injections, or during surgery via contaminated medical instruments. Recently, investigators from the Centers for Disease Control and Prevention, Atlanta, Ga, and the California Department of Health Services, Berkeley, documented a large, unprecedented outbreak of community-acquired RGM infection, during which more than 100 patrons of a northern California nail salon contracted furunculosis in their legs as a result of exposure to whirlpool footbaths that were contaminated with M fortuitum. OBSERVATIONS: We report the clinical and epidemiological findings in 3 cases of lower extremity RGM infections that occurred after similar whirlpool footbath exposure at several different nail salons in southern California. These infections typically presented as recurrent furunculosis, causing considerable morbidity as a result of scarring, delayed diagnosis, and the need for long-term polymicrobial therapy. CONCLUSIONS: Rapidly growing mycobacterial infections related to pedicures may continue to occur in a sporadic fashion. Clinicians should consider the possibility of RGM infection and inquire about recent pedicures in a patient with recurrent lower extremity furunculosis and abscesses that are unresponsive to conventional antibiotic therapy. PMID- 12756101 TI - Telepathology in the diagnosis of routine dermatopathologic entities. AB - BACKGROUND: Telepathology involves the use of video technology to facilitate remote-site diagnosis. To our knowledge, no studies have compared the reproducibility of real-time telepathology between dermatopathologists with that of traditional 2-headed microscopy in the diagnoses of routine dermatopathologic entities. OBSERVATIONS: The kappa statistic for both techniques was favorable: 0.76 (telepathology) vs 0.93 (conventional 2-headed microscopy); P =.04. The time taken per case was 42 seconds (telepathology) vs19 seconds (conventional 2 headed microscopy); P =.003. CONCLUSIONS: Telepathology between 2 remote diagnostic centers offers a feasible means of facilitating the remote-site diagnosis of routine dermatopathologic entities. Although diagnostic accuracy and time taken per case were acceptable with video-assisted diagnosis, conventional microscopy had significantly higher accuracy and shorter time per diagnosis. PMID- 12756102 TI - Basal cell carcinoma with pulmonary and lymph node metastasis causing death. AB - BACKGROUND: The incidence of metastatic basal cell carcinoma ranges from 0.003% to 0.55%. The 230 reported cases most often occurred in long-standing recurrent lesions and appeared in regional nodes or the lungs. OBSERVATIONS: The stromal dependence of the tumor provides an explanation for the nonmetastasizing nature of basal cell carcinoma. The dense fibrous stroma of the lymph node in the case of metastatic basal cell carcinoma reported in the present study is similar to other reported cases with metastases to lymph nodes, bone, bone marrow, glands, and subcutaneous tissue. CONCLUSIONS: This metastatic basal cell carcinoma demonstrated lymphatic and hematogenous dissemination to the lungs and lymph nodes. A dense accumulation of microvessels was present at the boundary of the tumor nests and dermal stroma and in the stroma surrounding the tumor in the lymph node. PMID- 12756103 TI - Childhood vitiligo and tacrolimus: immunomodulating treatment for an autoimmune disease. PMID- 12756104 TI - A 21-year-old woman with progressive reticulated pigmentation of the extremities. PMID- 12756105 TI - Large annular plaques on the trunk and arms. PMID- 12756106 TI - Acral and ear papules and arthritis. PMID- 12756108 TI - Narrowband UV-B phototherapy clears psoriasis through a combination of local and systemic effects. PMID- 12756107 TI - Brown macules symmetrically distributed on the neck, axillae, and thighs. PMID- 12756109 TI - On Zimmerman's "Dermatitis factitia as a war weapon". PMID- 12756110 TI - Hypothesis: a role for telomere crisis in spontaneous regression of melanoma. PMID- 12756111 TI - A new concept for acne therapy: a pilot study with zileuton, an oral 5 lipoxygenase inhibitor. PMID- 12756112 TI - Adults with atopic dermatitis and herpes simplex and topical therapy with tacrolimus: what kind of prevention? PMID- 12756113 TI - Immunohistochemical effects of temporary cessation of long-term acitretin treatment in keratinocytic intraepidermal neoplasia of renal transplant recipients. PMID- 12756114 TI - A systematic histologic analysis of nonablative laser therapy in a porcine model using the pulsed dye laser. AB - BACKGROUND: To our knowledge, no systematic analysis of nonablative laser therapy has been performed. OBJECTIVE: To alter the parameters (fluence, spot size, pulse duration, and use of cooling spray) for the pulsed dye laser to determine the precise settings that would yield the most favorable dermal remodeling in a porcine model. METHODS: Research was conducted in an animal laboratory at Albany Medical College. An anesthetized Yucatan miniature pig was subjected to a pulsed dye laser at various parameters. After 10 weeks, the laser-treated areas were harvested and processed for blinded, randomized, histologic evaluation. Negative (nontreated skin) and positive (ablative carbon dioxide laser-treated skin) controls were compared with the nonablative pulsed dye laser-treated areas. MAIN OUTCOME MEASURES: Quantitative assessment of collagen band width and cells per high-power field and qualitative assessment of epidermal and dermal changes. RESULTS: A significant difference (P<.001) in collagen band width was evident when nonablative laser-treated skin and carbon dioxide ablative laser-treated skin specimens were compared with untreated skin specimens, but no significant (P =.18) difference existed between the nonablative and ablative modalities. Similarly, cellular hypertrophy, as measured by high-power field, corroborated the previous findings. Furthermore, a higher fluence, a larger spot size, and a longer pulse duration proved statistically significant for increased collagen band width (P =.01, P<.001, and P<.001, respectively), and a larger spot size and a longer pulse duration exhibited significance for cells per high-power field (P =.02 and P =.009, respectively), with a trend toward significance for higher fluence (P =.09). Overall, the dermis was considerably thicker for nonablative and ablative laser-treated areas compared with untreated skin, but this could not be quantified because the depth exceeded the punch biopsy instrument. The epidermis remained unchanged. CONCLUSIONS: The nonablative pulsed dye laser has demonstrated favorable histologic evidence of dermal remodeling, and its effects were similar to histologic changes seen with the carbon dioxide ablative laser, both of which were statistically significant compared with untreated skin, as seen in this preliminary animal model. PMID- 12756115 TI - Gore-Tex chin implants: a review of 324 cases. AB - Augmentation of the chin is a long-standing and effective technique for facial enhancement. We have used preformed expanded polytetrafluoroethylene (Gore-Tex) implants for chin augmentation for several years. For this study, we collectively pooled data detailing our experiences with this material. From January 1, 1998, to March 30, 2001, a total of 324 Gore-Tex chin implants were placed. No resorption or visible movement of any implant occurred. Two (0.62%) of the 324 implants became infected and were ultimately removed. No other complications occurred. This complication rate compares favorably to other reports. Five implants (1.5%) were removed or changed in size due to patient requests. All remaining patients (97.8%) were satisfied with their result. We also describe technical points and procedure modifications that have helped us achieve beneficial results for our patients. Gore-Tex is a reliable implant material that helps the surgeon to achieve a high degree of patient satisfaction in chin augmentation. PMID- 12756116 TI - A psychological profile of children with hemangiomas and their families. AB - OBJECTIVE: To assess the psychosocial impact of hemangiomas and their treatment on children with the disease and their families. DESIGN: Thirty-nine children who were treated for hemangiomas were examined by a questionnaire that addressed the emotional attitudes of the parent and child toward the disease and the related treatment. SETTING: Two private ambulatory surgery centers (in Latham and Charleston). RESULTS: Overall, the survey found a negative effect on the child's family, with considerable fear caused in part by adverse public commentary or attitudes--which was ameliorated by education from the primary care provider and specialist. However, the family's perception was that the child was not deeply affected by his or her condition and that treatment (laser, intralesional corticosteroids, oral corticosteroids, surgery, or a combination) did not change the child's emotional response to the disease. However, most parents observed that their child was too young to appreciate his or her malady. CONCLUSION: Given earlier intervention for children with late-involuting hemangiomas and the advent of more effective therapies, our survey did not seem to indicate that the children experienced significant emotional trauma from their condition; nevertheless, their families experienced appreciable emotional and psychological distress. PMID- 12756117 TI - Quantifying the arch position of the female eyebrow. AB - OBJECTIVE: To devise a quantitative aesthetic model for the eyebrow arch position (EAP) in women. METHODS: Full-face frontal magazine photographs of 100 fashion models published between January and July 2001 were analyzed. Apparent EAP relative to a line through the medial canthus parallel to the midline was compared with eyewidth (EW). A similar comparison was made between the lateral limbus (LL) and the EW. Standardized full-face frontal photographs of 105 randomly selected women aged 21 to 61 years were taken after obtaining informed consent. The photographs were analyzed in the same manner as those of the fashion models. Both populations had the medial and lateral extents of their eyebrows analyzed. RESULTS: The mean +/- SD EAP:EW ratio for the fashion models was 0.978 +/- 0.131; the mean +/- SD LL:EW ratio was 0.735 +/- 0.0673. The mean +/- SD EAP:EW ratio for the randomly selected group was 0.929 +/- 0.146; the mean +/- SD LL:EW ratio was 0.762 +/- 0.0420. CONCLUSIONS: The EAP has been described as being above the LL. This does not reflect the EAP seen in both of our study groups. The EAP seems to be 93% to 98% of an EW in the aesthetic model derived from these data. PMID- 12756118 TI - Dorsal onlay cartilage autografts: comparing resorption in a rabbit model. AB - OBJECTIVE: To compare the resorption characteristics of dorsal onlay cartilage autografts from the septum, auricle, and rib. METHODS: Fourteen New Zealand white rabbits underwent harvesting of equal-sized septal, auricular, and costal cartilage grafts free of perichondrium. All autografts were implanted subcutaneously on the nasal dorsum and then removed after 3 months. Graft mass, chondrocyte density, and histologic features from hematoxylin-eosin-stained sections were compared before and 3 months after implantation. RESULTS: At 3 months after implantation, septal cartilage grafts averaged 30.8% resorption by weight, followed by auricular (23.1%) and costal (7.6%) cartilage. All 3 groups demonstrated similar changes in chondrocyte density and minor calcification at 3 months. There was no evidence of necrosis or inflammatory changes in any of the specimens. CONCLUSIONS: Although the septum is often the preferred source of autogenous cartilage for nasal reconstruction, short-term resorption of septal cartilage appears to be higher for dorsal onlay grafts. The low resorption of costal cartilage may be due in part to its compact shape compared with septal and auricular cartilage. It remains to be seen whether these differences in resorption persist in the long term. PMID- 12756119 TI - Endoscopically assisted repair of subcondylar fractures of the mandible: an evolving technique. AB - OBJECTIVE: To review one surgeon's experience with the endoscopic approach to assist with reduction and rigid fixation of subcondylar fractures of the mandible. DESIGN: Chart review of all cases in which endoscopic techniques were used to assist with the reduction and, when possible, repair of these fractures. The numbers of fractures approached, successfully repaired with the use of plates and screws, and not successfully plated with this approach were documented. SETTING: All surgeries were performed in the operating room at a university hospital. PATIENTS: All patients who underwent endoscopic exploration of a single or bilateral subcondylar fractures of the mandible were included. Seventeen explorations were carried out in 12 patients (age range, 16-39 years). Associated mandibular and other facial fractures were noted. INTERVENTIONS: Endoscopic exploration via a transoral approach. A secondary port in the submandibular region was made in 13 of 17 fracture explorations, and this second port was used primarily for the application of downward traction on the angle of the mandible. Plates were introduced transorally, while screws were placed through a transbuccal trochar. MAIN OUTCOME MEASURE: Success was judged by the successful reduction of the fracture and application of a rigid fixation plate by means of the limited, endoscopically assisted approach, a measure of the ability to accomplish the procedure, not an evaluation of functional results. (With these criteria, 9 of 10 successes had normal function at last follow-up, as did the 10th after revision.) RESULTS: Rigid plate fixation was completed endoscopically for 10 fractures, and 2 were plated after conversion to a full open approach. Four were reduced but could not be plated, and in 1 exploration, a bent plate was removed, but a new plate was not applied. Nine of the 10 fractures plated endoscopically resulted in normal occlusion and function. In the 10th case, a persistent malocclusion necessitated reexploration and refixation, resulting in a successful functional outcome with normal occlusion. CONCLUSION: The endoscopically assisted approach for the repair of subcondylar fractures of the mandible is a feasible but challenging technique. PMID- 12756120 TI - The use of "inside-out" lateral osteotomies to improve outcome in rhinoplasty. AB - OBJECTIVE: To evaluate the anatomic effects of the transnasal nasal "inside-out" lateral osteotomy in comparison to a continuous lateral osteotomy to widen the excessively narrow bony nasal pyramid. DESIGN: A series of 4 fresh cadavers were used. Lateral osteotomies were performed on each cadaver. On one side, a continuous lateral osteotomy was performed with a curved, guarded 4-mm osteotome. On the contralateral side, a straight 2-mm osteotome was used to perform the transnasal perforating inside-out lateral osteotomy. The soft tissue was then dissected off the nasal dorsum and the effects were noted. Specifically, the following factors were assessed: (1) the continuity of the internal and external periosteum; (2) any change, without manipulation, of the position of the lateral nasal walls; (3) the mobility of the fragments; and (4) the effect of any of the preceding factors on the nasal airway. RESULTS: All 4 cadavers showed identical results. The external and internal periostea were significantly disrupted on each of the sides with continuous osteotomies. The periosteum was completely preserved on the sides, which underwent inside-out lateral osteotomies, with the exception of the 3 or 4 holes created by the 2-mm osteotome. In each of the sides with continuous osteotomies, the lateral nasal wall was demonstrably unstable, as the segment was, to varying degrees, displaced inwardly. The inside-out lateral osteotomized segments were uniformly lateralized and stable to gentle palpation. Finally, these changes corresponded to a visible effect on the diameter of the nose in the region of the nasal valve. The inward displacement of the segments undergoing continuous osteotomies narrowed the airway. However, the contralateral inside-out osteotomized segments were lateralized, which widened the diameter of the valve. CONCLUSIONS: The inside-out lateral osteotomy is an effective technique for lateral repositioning of the bony lateral sidewall of the nose. It is reproducible and accurate and appears to provide greater preservation of the periosteal support of the bony segments than a continuous osteotomy. This technique provides a more predictable long-term result, with preservation and/or improvement of the nasal airway. The procedure is ideal for managing the bony nasal segment that needs lateralization, particularly in revision rhinoplasties or after trauma. PMID- 12756121 TI - Orbital volume augmentation for late enophthalmos using the deep lateral wall. AB - Orbital volume augmentation to address enophthalmos and hollowing of the superior sulcus has been described with a variety of materials and from a variety of approaches.(1-4) A common location for volume augmentation is the inferomedial orbital wall; this surface is often the one that was expanded related to orbital trauma, and it is easily accessed through hidden conjunctival or caruncular incisions. PMID- 12756122 TI - The coronal incision: sinusoidal, sawtooth, and postauricular techniques. AB - The coronal incision is a popular and versatile surgical approach for access to the cranial vault and the upper two thirds of the facial skeleton. It provides excellent exposure to allow neurosurgical access, craniofacial osteotomies, repair of facial fractures, calvarial bone grafting, and cosmetic procedures such as the forehead lift. Since the introduction and acceptance of the coronal approach, a variety of modifications of the incision have been used, including methods for camouflaging the incision in the hair. To quickly and easily produce a sinusoidal or sawtooth coronal incision line with or without postauricular extension, a tape measure is positioned at the level of the anteriormost point of the auricular helix. A mark is made every 2 cm on alternating sides of the tape measure. The tape measure is advanced coronally until the vertex of the skull is reached, and the procedure is repeated on the contralateral side. After the tape measure is removed, the resulting regularly spaced marks are connected in a sinusoidal or a sawtooth fashion. A postauricular incision can be easily extended from either the sinusoidal or sawtooth coronal template. PMID- 12756123 TI - Reconstruction of congenital and acquired auricular scaphal deficiencies. AB - Although complications from piercing of the helical rim are unusual, infections in this area after piercing can produce devastating changes in the cartilaginous framework. Scaphal height is also reduced in certain congenital pinna anomalies. We developed a method of scaphal reconstruction that uses a composite graft harvested from the ipsilateral conchal bowl. The conchal bowl defect is repaired with an ipsilateral full-thickness graft of postauricular skin. This technique restores scaphal height and helical contour with minimal donor site morbidity. PMID- 12756124 TI - Prophylactic lateral canthopexy in lower blepharoplasties. AB - Lower eyelid malposition is a known complication of lower (lid) blepharoplasty surgery. The prevention of this complication is easier than its treatment. Over the past 5 years in my practice, 247 patients have had lower blepharoplasties with a canthopexy procedure and in some cases a tarsal strip canthoplasty. The criterion for a lateral tarsal strip canthoplasty was a lid distraction distance greater than or equal to 10 mm. Lid distraction distance is the distance the eyelid can be pulled away from the globe after the initial skin incision has been made and measured with calipers. If the eyelid can be pulled away from the globe less than 10 mm, then a canthopexy is performed, which occurred in 98% of cases. Patients had lateral canthopexies regardless of age or preoperative assessment. Herein, I describe a simple method of canthopexy that can be performed on most patients having a lower blepharoplasty, to not only achieve a cosmetically superior result but also to prevent eyelid malposition or ectropion. PMID- 12756125 TI - Review of prophylactic lateral canthopexy in lower blepharoplasties. PMID- 12756126 TI - Johann Friedrich Dieffenbach: the many-sided Odysseus. PMID- 12756128 TI - Familial hemiplegic migraine. PMID- 12756129 TI - Neurological aspects of taste disorders. PMID- 12756130 TI - Application of microarrays to neurological disease. PMID- 12756131 TI - Expanding the phenotypic spectrum of the CACNA1A gene T666M mutation: a description of 5 families with familial hemiplegic migraine. AB - BACKGROUND: Familial hemiplegic migraine (FHM) is a rare autosomal dominant subtype of migraine with aura. Missense mutations in the chromosome 19 CACNA1A calcium channel gene have been found in approximately half of the families. The T666M mutation, replacing a threonine by a methionine at residue number 666, is the most frequent mutation, reported in 14 independent FHM families; other mutations have so far been described in only 1 or 2 families each. The clinical features of T666M families have been reported, but the course is unknown. OBJECTIVE: To present a detailed description of the clinical features of new FHM families in which we identified the T666M mutation in our CACNA1A screening program. METHODS: As part of our ongoing genetic screening, mutation analysis of the CACNA1A gene was performed by single-strand conformational polymorphism analysis in 33 probands of families with FHM. RESULTS: We identified the T666M mutation in 5 unrelated FHM families. In 3 of the families, patients displayed cerebellar ataxia. In 1 family, some affected members with the mutation had attacks with confusion but without hemiparesis. In 1 family, patients had progressive cognitive dysfunction. CONCLUSIONS: The T666M mutation is the most frequent CACNA1A mutation in FHM; it was found in 5 of 33 FHM families at our laboratory, and in 19 of 39 families with a known mutation reported in the literature (including the present study). Screening for the T666M mutation should therefore be the first step when screening families with FHM. There is a remarkable clinical heterogeneity among families with the T666M mutation. PMID- 12756132 TI - Subthalamic stimulation in Parkinson disease: intraoperative predictive factors. AB - BACKGROUND: High-frequency stimulation of the subthalamic nucleus (STN) is an effective treatment for advanced forms of Parkinson disease. Postoperative improvement of motor parkinsonian disability is known to depend on patient selection and surgical targeting. OBJECTIVE: To determine which clinical and electrophysiological variables evaluated during the operation predict the postoperative clinical outcome of patients with Parkinson disease treated by bilateral high-frequency stimulation of the STN. METHODS: Intraoperative clinical and electrophysiological data obtained in 41 patients with Parkinson disease who underwent bilateral implantation of electrodes for STN stimulation were correlated with the improvement in parkinsonian disability assessed 6 months after the operation. RESULTS: The extent of STN neuronal activity recorded along the trajectory of the therapeutic electrode had no effect on the postoperative clinical outcome. The intraoperative improvement in segmental akinesia, but not rigidity, was predictive of the postoperative improvement in parkinsonian motor disability and reduction in daily levodopa-equivalent dosage. Parkinsonian motor disability scores assessed after surgery were lower in patients with intraoperative stimulation-induced dyskinesias than in those without stimulation induced dyskinesias. CONCLUSION: The improvement of segmental akinesia and the observation of dyskinesias provoked by stimulation during the operation predict the best postoperative effects of bilateral STN stimulation on parkinsonian motor disability. PMID- 12756133 TI - Novel tau polymorphisms, tau haplotypes, and splicing in familial and sporadic frontotemporal dementia. AB - BACKGROUND: A subset of familial cases (FTDP-17) of frontotemporal dementia (FTD) are caused by mutations in the tau gene. The role of tau gene mutations and haplotypes in sporadic FTD and the functional consequences of tau polymorphisms are unknown. OBJECTIVES: To investigate (1) the frequency of known FTDP-17 mutations in familial and sporadic FTD and compare these results with previous studies; (2) whether the tau H1 haplotype is associated with FTD; and (3) the functional effect of intronic tau sequence variations. PATIENTS AND METHODS: Patients with familial and sporadic FTD were screened for mutations in the microtubule-binding region of tau. The frequencies of tau haplotypes and genotypes were compared between patients with FTD and control subjects. We analyzed the splicing effect of novel intronic polymorphisms associated with FTD. RESULTS: The P301L mutation was detected in 11% of familial FTD cases. The H1 haplotype was not overrepresented in patients with FTD, but the P301L mutation appeared on the background of the H2 tau haplotype. We identified 4 novel single nucleotide polymorphisms in intron 9 and a 9-base pair deletion in intron 4A. A C to-T transition 177 base pairs upstream from exon 10 was significantly increased in patients with FTD compared with controls. Direct analysis of brain tissue from a patient with this variant showed an increase in exon 10-containing tau transcripts. CONCLUSIONS: Sequence variations in intronic or regulatory regions of tau may have previously unrecognized consequences leading to tau dysfunction and neurodegeneration. PMID- 12756134 TI - Incipient CADASIL. AB - BACKGROUND: Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is caused by mutations in the NOTCH3 gene. Knowledge of disease expression in young adult NOTCH3 mutation carriers (MCs) is limited. OBJECTIVE: To characterize clinical, neuropsychological, and radiological status in NOTCH3 MCs younger than 35 years. DESIGN: Clinical characterization and blinded survey comparing MCs with non-MCs. SETTING: Referral center. PARTICIPANTS: Individuals younger than 35 years who were at a 50% risk of a NOTCH3 mutation, from our CADASIL database. Thirteen individuals, from 8 families, met the criteria. METHODS: Comprehensive clinical, genetic, neuropsychological, and radiological investigations. Magnetic resonance images were scored according to a standardized white matter hyperintensities rating scale. RESULTS: Six individuals, from 5 families, were MCs. Clinical symptoms consisted of migraine (with aura), stroke, and stroke-like episodes. We did not find evidence for psychiatric disturbances, functional disability, or cognitive dysfunction, compared with non-MCs. Radiologically, a characteristic magnetic resonance imaging lesion pattern emerged for all MCs. This comprised white matter hyperintensities in the anterior temporal lobes, the frontal lobes, and the periventricular frontal caps. CONCLUSIONS: Migraine (with aura) and stroke can present in NOTCH3 MCs younger than 35 years; however, more importantly, physical function and cognition are intact. Possible subtle cognitive dysfunction needs to be assessed in a larger study. White matter hyperintensities on magnetic resonance imaging are characteristic, and are consistently visualized from the age of 21 years and onward. Awareness of the clinical and radiological features of CADASIL in those younger than 35 years should increase early diagnosis and allow for customized counseling of young adults from families with CADASIL. PMID- 12756135 TI - Young-onset Parkinson disease with and without parkin gene mutations: a fluorodopa F 18 positron emission tomography study. AB - BACKGROUND: Mutations of the parkin gene are frequently encountered in patients with young-onset Parkinson disease (YOPD), but the effects of this mutation on the nigrostriatal dopaminergic degeneration are not well established. OBJECTIVE: To analyze, using positron emission tomography and fluorodopa F 18, the severity and profile of striatal dopaminergic metabolism in YOPD patients with and without parkin gene mutations. METHODS: We performed positron emission tomography with fluorodopa F 18 in 19 YOPD patients with parkin gene mutations (parkin patients), 6 YOPD patients without parkin gene mutations (nonparkin patients), and 9 healthy controls. Putamen and caudate nucleus fluorodopa F 18 uptake was assessed using regions of interest analysis. RESULTS: In parkin patients, the striatal fluorodopa F 18 uptake reduction was 36.3%, 51.3%, and 66.7%, respectively, for the caudate nucleus, anterior putamen, and posterior putamen compared with controls. In nonparkin patients, this reduction was 23.0%, 43.6%, and 73.0%, respectively. This reduction was asymmetrical according to the most affected hemibody for the anterior and posterior putamen in parkin patients and for the posterior putamen in nonparkin patients. A rostrocaudal gradient was observed with a severe decrease in fluorodopa F 18 uptake in the putamen and relative sparing of the caudate nucleus. There was no significant difference of striatal fluorodopa F 18 uptake between our 2 YOPD populations. In parkin patients, no significant correlation was found among fluorodopa F 18 uptake, motor disability, and the type of mutations. In nonparkin patients, there was a significant correlation between fluorodopa F 18 uptake and clinical severity. CONCLUSIONS: The pattern of fluorodopa F 18 uptake in the striatum of YOPD patients is similar to that of patients with idiopathic Parkinson disease and does not depend on the presence or absence of mutations of the parkin gene. PMID- 12756136 TI - A common NURR1 polymorphism associated with Parkinson disease and diffuse Lewy body disease. AB - BACKGROUND: NURR1 plays a key role in mesencephalic dopaminergic neuron development and survival. A homozygous NURR1 polymorphism (a single base-pair insertion in intron 6) (NI6P) has been reported to be associated with Parkinson disease (PD). OBJECTIVE: To assess the association of the NI6P with PD and diffuse Lewy body disease. DESIGN: Case-control study. SETTING: Movement disorders clinic and tissue provided by brain banks. PATIENTS: Patients with pathologically proven PD (n = 37) or diffuse Lewy body disease (n = 35), neuropathologically normal control subjects (n = 59), those clinically diagnosed as having PD (n = 66), and spousal controls (n = 29). METHODS: Determining the frequency of heterozygotes and homozygotes for the NI6P by DNA sequencing and restriction endonuclease analyses. RESULTS: Overall, 41 (39.8%) of the 103 patients with PD were heterozygotes compared with 22 (25.0%) of the 88 controls (P =.03), with a relative risk (estimated from the odds ratio) for PD of 2.03 (95% confidence interval, 1.08-3.81) for heterozygotes vs wild type subjects. Heterozygotes were more frequent in the subgroup of patients with pathologically confirmed PD (18 [48.6%] of 37) vs controls (14 [23.7%] of 59) (P =.01), with a relative risk for PD of 2.84 (95% confidence interval, 1.17-6.88) for heterozygotes vs wild type subjects. In patients clinically diagnosed as having PD, heterozygotes were more frequent in early-onset cases (onset at < or =45 years) (10 [55.6%] of 18) compared with late-onset cases (onset at >45 years) (10 [23.8%] of 42) (P =.02) or spousal controls (8 [27.6%] of 29) (P =.06), with a relative risk for early-onset PD of 4.17 (95% confidence interval, 1.13-15.33) for heterozygotes vs subjects with 2 wild type alleles. The homozygous NI6P was not associated with PD, but was present in 6 (17.1%) of the 35 patients with diffuse Lewy body disease compared with 3 (5.1%) of the 59 controls (P =.06). CONCLUSIONS: The common heterozygous NI6P is associated with an increased risk of PD. An association of borderline significance was found for the homozygous NI6P and diffuse Lewy body disease. PMID- 12756137 TI - Neurofibrillary tangles, amyloid, and memory in aging and mild cognitive impairment. AB - BACKGROUND: Large numbers of neurofibrillary tangles (NFTs) and amyloid plaques are diagnostic markers for Alzheimer disease (AD), but lesser numbers of these lesions are also seen in nondemented elderly individuals. Much of the existing literature suggests that the NFTs of AD have a closer correlation with cognitive function than do amyloid plaques. Whether a similar relationship exists in normal aging and mild cognitive impairment (MCI), a condition that frequently reflects a preclinical stage of AD, remains unknown. OBJECTIVE: To determine the distribution patterns of beta-amyloid plaques and NFTs and the association of these lesions with memory performance in nondemented individuals. METHODS: We investigated regional distributions and neuropsychological correlates of NFTs and amyloid plaques in cognitively normal elderly persons and subjects with MCI who received neuropsychological testing before death. Subjects Eight nondemented subjects who volunteered to receive annual neuropsychological testing and agreed to brain donation were studied. Five subjects showed no cognitive impairment, and 3 were diagnosed with MCI. RESULTS: Distribution of NFTs followed a rigorous and hierarchical pattern, but distribution of amyloid plaques varied among individuals. Subjects with MCI displayed higher NFT densities than did nonimpaired subjects. In addition, NFT density in the temporal lobe correlated with memory scores, whereas density of amyloid plaques did not. CONCLUSIONS: Neurofibrillary tangles are more numerous in medial temporal lobe regions associated with memory function and show a relationship to performance on memory tests in nondemented individuals. These results suggest that NFTs may constitute a pathological substrate for memory loss not only in AD but also in normal aging and MCI. PMID- 12756138 TI - Molecular diagnosis and prophylactic therapy for presymptomatic Chinese patients with Wilson disease. AB - BACKGROUND: The potential for therapy for Wilson disease (WD) emphasizes the importance of presymptomatic diagnosis in families with WD (WD families). OBJECTIVES: To investigate the feasibility of presymptomatic DNA diagnosis and evaluate the efficacy of zinc sulfate therapy in WD families. METHODS: Seventy eight clinically unaffected siblings were studied from 51 unrelated WD families that were ascertained by affected individuals. The diagnosis in presymptomatic patients was established by a combination of direct mutational analysis and haplotype analysis with 3 short tandem repeat markers. The presymptomatic patients were treated with 50 mg of elemental zinc sulfate twice a day from the time of molecular diagnosis and followed up for 3 to 5 years. RESULTS: Of the 78 siblings, 17 were diagnosed as presymptomatic patients. Kayser-Fleischer rings were absent in 7 and faint in 4 of the 17 presymptomatic patients. The serum ceruloplasmin values gradually increased and 24-hour urinary copper values gradually diminished during zinc therapy, which indicate effective control of copper metabolism. None of the siblings developed clinical symptoms of WD or adverse effects from zinc therapy. CONCLUSION: We conclude that presymptomatic DNA diagnosis and zinc therapy are effective treatment of patients with WD. PMID- 12756139 TI - POEMS syndrome associated with ischemic stroke. AB - BACKGROUND: A syndrome variously combining peripheral neuropathy, visceromegaly, endocrinopathy, monoclonal gammopathy, and skin changes (POEMS syndrome) is a rare variant of plasma cell dyscrasia with multisystemic manifestations. Acute ischemic strokes in patients with POEMS syndrome have rarely been reported, and the pathophysiologic mechanism of this disease is unknown. Fibrinogen is reported to be an independent risk factor for cerebrovascular disease and is correlated with the interleukin 6 level in the plasma. The serum level of interleukin 6 is high in the active stage of POEMS syndrome. OBJECTIVE: To describe the neuroimaging findings and fibrinogen levels in patients with POEMS syndrome. DESIGN: Case series. SETTING: The neurology department of a tertiary referral center. METHODS: Three patients with an acute cerebral infarction associated with POEMS syndrome underwent magnetic resonance imaging, diffusion-weighted imaging, magnetic resonance angiography, transcranial Doppler ultrasonography, and serum fibrinogen level and serum C-reactive protein level analysis. The serum fibrinogen level before the stroke was collected retrospectively from the hospital medical records. RESULTS: There was an elevated fibrinogen level in all of the patients. In 2 patients, unilateral or bilateral end artery border-zone infarcts were observed on the brain magnetic resonance imaging scan. The serum fibrinogen level was high before the stroke in 2 patients. CONCLUSIONS: The POEMS syndrome can be associated with stroke, particularly end artery border-zone infarctions. We suggest that an elevated fibrinogen level might play a role in the pathogenesis of stroke. PMID- 12756140 TI - Depression as a risk factor for Alzheimer disease: the MIRAGE Study. AB - BACKGROUND: Depression symptoms may be associated with the development of Alzheimer disease (AD). OBJECTIVES: To evaluate the association between depression symptoms and risk of AD, and to explore the temporal aspects of this association. SETTING: Academic institutions with specialized memory clinics. DESIGN: Cross-sectional, family-based, case-control study with standardized self- and proxy questionnaires to collect information on depression symptoms and other risk factors. PARTICIPANTS: A total of 1953 subjects with AD and 2093 of their unaffected relatives enrolled in the Multi-institutional Research in Alzheimer's Genetic Epidemiology Study. MAIN OUTCOME MEASURES: Odds ratios (ORs) of AD were estimated with and without depression symptoms, adjusted for age, sex, education, history of head trauma, and apolipoprotein E status. RESULTS: There was a significant association between depression symptoms and AD (adjusted OR, 2.13; 95% confidence interval [CI], 1.71-2.67). In families where depression symptoms first occurred within 1 year before the onset of AD, the association was higher (OR, 4.57; 95% CI, 2.87-7.31), while in the families where the depression symptoms first occurred more than 1 year before the onset of AD, the association was lower (OR, 1.38; 95% CI, 1.03-1.85). In families where depression symptoms first occurred more than 25 years before the onset of AD, there was still a modest association (OR, 1.71; 95% CI, 1.03-2.82). CONCLUSIONS: Depression symptoms before the onset of AD are associated with the development of AD, even in families where first depression symptoms occurred more than 25 years before the onset of AD. These data suggest that depression symptoms are a risk factor for later development of AD. PMID- 12756141 TI - Congenital myasthenic syndrome with episodic apnea in patients homozygous for a CHAT missense mutation. AB - BACKGROUND: The syndrome of congenital myasthenia with episodic apnea (CMS-EA) was previously found to be due to mutations in the choline acetyltransferase gene (CHAT). OBJECTIVE: To identify the mutations underlying CMS-EA in a Turkish multiplex family. DESIGN: Direct sequencing of the CHAT gene. PATIENTS: A consanguineous Turkish family with 2 siblings affected by muscular weakness and episodic respiratory distress. RESULTS: The sequencing of CHAT coding exons identified a previously unknown missense mutation that affected a highly conserved amino acid residue (I336T). The mutation was absent in 164 control chromosomes. CONCLUSIONS: The high degree of conservation in different species strongly suggests that I336T is a functionally important amino acid residue. The absence of I336T from a large control sample further supports the pathogenic role of I336T in CMS-EA. This is the second report of CHAT mutations causing presynaptic CMS. PMID- 12756142 TI - Adult polyglucosan body disease associated with lewy bodies and tremor. AB - BACKGROUND: Adult polyglucosan body disease (PGBD) is rare and typically presents with upper and lower motor neuron involvement and neurogenic bladder. Extrapyramidal features are unusual in PGBD and are presumed secondary to widespread pathology that includes the basal ganglia. There are no prior reports of Lewy bodies in PGBD. OBJECTIVE: To report a unique finding of Lewy bodies in a patient with PGBD. REPORT OF A CASE A 46-year-old woman initially presented with a 4-year history of resting tremor. The tremor responded to levodopa therapy. Several months later, she developed upper and lower motor neuron involvement and other clinical features of PGBD. A sural nerve biopsy specimen revealed intra axonal polyglucosan bodies that confirmed the clinical diagnosis. Bulbar and limb weakness progressed, and she developed dementia. She died 6 years after onset. At autopsy, extensive polyglucosan body formation was found in many regions of the central nervous system. In addition, numerous alpha-synuclein staining Lewy bodies were observed in the substantia nigra, accompanied by marked neuron depopulation. CONCLUSIONS: To our knowledge, this is the first report of adult PGBD associated with Lewy bodies and levodopa-responsive tremor. Although polyglucosan bodies were seen in substantia nigra, it is most likely that our patient had coexisting Parkinson disease. PMID- 12756143 TI - Sporadic Creutzfeldt-Jakob disease mimicking variant Creutzfeldt-Jakob disease. AB - BACKGROUND: The determination of the form of prion disease and early diagnosis are important for prognostic, public health, and epidemiologic reasons. OBJECTIVE: To describe a patient with sporadic Creutzfeldt-Jakob disease (sCJD) who had a clinical history and initial electroencephalogram and magnetic resonance imaging findings consistent with variant CJD (vCJD). RESULTS: Results of a repeated electroencephalogram were suggestive of sCJD, and a subsequent brain biopsy confirmed this diagnosis. CONCLUSIONS: This case cautions against relying solely on T2- and diffusion-weighted pulvinar hyperintensity and clinical features to differentiate between vCJD and sCJD, and further supports established diagnostic criteria for vCJD. PMID- 12756145 TI - Should the multicenter carotid endarterectomy trials be repeated? PMID- 12756144 TI - Tolterodine and memory: dry but forgetful. AB - BACKGROUND: Anticholinergic drugs are known to produce or enhance cognitive deficits. Tolterodine tartrate is marketed as a bladder-selective anticholinergic drug that is reported to be free of significant cognitive adverse effects. OBJECTIVE: To describe a 46-year-old woman who had memory loss and abnormal memory test results that improved when she discontinued tolterodine therapy. RESULTS: While taking tolterodine, the patient's score on the delayed free recall portion of the Hopkins Verbal Learning Test-Revised was at the first percentile. One month after discontinuing tolterodine therapy, this test was administered a second time using an alternative form and she showed marked improvement scoring above the 75th percentile. CONCLUSIONS: Tolterodine therapy caused cognitive dysfunction in our patient. It is possible that cognitive dysfunction is a common result of tolterodine treatment, but in the absence of testing, remains undiagnosed. Alternatively, our patient may have had aberrant metabolism of this drug or an increased sensitivity as a result of incipient Alzheimer disease. PMID- 12756146 TI - Resolved: NASCET and ACAS need not be repeated: the affirmative position. PMID- 12756147 TI - How should we design studies for stroke prevention? PMID- 12756148 TI - History of dermatomyositis. PMID- 12756149 TI - Cervical epidural hematoma causing hemiparesis. PMID- 12756150 TI - Probing the contribution of IKs to canine ventricular repolarization: key role for beta-adrenergic receptor stimulation. AB - BACKGROUND: In large mammals and humans, the contribution of IKs to ventricular repolarization is still incompletely understood. METHODS AND RESULTS: In vivo and cellular electrophysiological experiments were conducted to study IKs in canine ventricular repolarization. In conscious dogs, administration of the selective IKs blocker HMR 1556 (3, 10, or 30 mg/kg PO) caused substantial dose-dependent QT prolongations with broad-based T waves. In isolated ventricular myocytes under baseline conditions, however, IKs block (chromanols HMR 1556 and 293B) did not significantly prolong action potential duration (APD) at fast or slow steady state pacing rates. This was because of the limited activation of IKs in the voltage and time domains of the AP, although at seconds-long depolarizations, the current was substantial. Isoproterenol increased and accelerated IKs activation to promote APD95 shortening. This shortening was importantly reversed by HMR 1556 and 293B. Quantitatively similar effects were obtained in ventricular-tissue preparations. Finally, when cellular repolarization was impaired by IKr block, IKs block exaggerated repolarization instability with further prolongation of APD. CONCLUSIONS: Ventricular repolarization in conscious dogs is importantly dependent on IKs. IKs function becomes prominent during beta-adrenergic receptor stimulation, when it promotes AP shortening by increased activation, and during IKr block, when it limits repolarization instability by time-dependent activation. Unstimulated IKs does not contribute to cellular APD at baseline. These data highlight the importance of the synergism between an intact basal IKs and the sympathetic nervous system in vivo. PMID- 12756151 TI - Adjuvant treatment with a glycoprotein IIb/IIIa receptor inhibitor increases the therapeutic window for low-dose tissue plasminogen activator administration in a rat model of embolic stroke. AB - BACKGROUND: Platelet aggregation and fibrin deposition are key events leading to microvascular thrombosis and progressive impairment of downstream microvascular perfusion after stroke. We tested the hypothesis that inhibition of platelet function with a GP IIb/IIIa receptor antagonist would increase the efficacy and safety and increase the time window for thrombolytic therapy for stroke with full and half-dose tissue plasminogen activator (tPA). METHODS AND RESULTS: Rats were subjected to embolic middle cerebral artery occlusion. Four hours after ischemia, rats were treated with 7E3 F(ab')2 (6 mg/kg) in combination with tPA at doses of 10 and 5 mg/kg, tPA alone at a dose of 10 or 5 mg/kg, 7E3 F(ab')2 (6 mg/kg) alone, or saline. Combination treatment with 7E3 F(ab')2 and tPA (full- or half dose) significantly (P<0.05) reduced infarct volume and neurological deficits compared with saline-treated rats. However, treatment with 7E3 F(ab')2 or tPA (full- or half-dose) alone did not reduce infarct volume. Quantitative measurements of cerebral microvessels perfused by FITC-dextran revealed that combination treatment with 7E3 F(ab')2 and full-dose tPA significantly (P<0.05) increased the percentage of FITC-dextran-perfused vessels compared with saline and full-dose tPA-treated rats. In addition, treatment with 7E3 F(ab')2 in combination with full-dose tPA significantly (P<0.05) decreased microvascular platelet accumulation and matrix metalloproteinase 9 immunoreactivity and protected against loss of collagen IV immunoreactivity. CONCLUSIONS: Combination treatment with 7E3 F(ab')2 with full- and half-dose tPA at 4 hours after ischemia significantly reduces infarct volume and improves neurological outcome. Enhancement of patency and integrity of cerebral microvessels most likely contributes to the benefits observed with this combination therapy. PMID- 12756152 TI - Role of L-type calcium channels in pacing-induced short-term and long-term cardiac memory in canine heart. AB - BACKGROUND: We tested the hypothesis that ICa,L is important to the development of cardiac memory. METHODS AND RESULTS: The effects of L-type Ca2+ channel blockade and beta-blockade were tested on acutely anesthetized and on chronically instrumented, conscious dogs. Short-term memory (STM) was induced by 2 hours of ventricular pacing and long-term memory (LTM) by ventricular pacing for 21 days. STM dogs received placebo, nifedipine, or propranolol, and LTM dogs received placebo, atenolol, or amlodipine. AT1 receptor blockade (candesartan) and ACE inhibition (trandolapril) were also tested in LTM. Microelectrodes were used to record transmembrane potentials from isolated epicardial and endocardial slabs using a protocol simulating STM in intact animals. Left ventricular epicardial myocytes from LTM or sham control dogs were dissociated, and ICa,L was recorded (whole-cell patch-clamp technique). Evolution of STM and LTM was attenuated by ICa,L blockers but not beta-blockers. Neither AT1 receptor blockade nor ACE inhibition suppressed LTM. In microelectrode experiments, pacing induced an epicardial-endocardial gradient change mimicking STM that was suppressed by nifedipine. In patch-clamp experiments, peak ICa,L density in LTM and control were equivalent, but activation was more positive and time constants of inactivation longer in LTM (P<0.05). CONCLUSIONS: ICa,L blockade but not beta adrenergic blockade suppresses cardiac memory. LTM evolution is unaffected by angiotensin II blockade and is associated with altered ICa,L kinetics. PMID- 12756153 TI - Phased-array intracardiac echocardiography monitoring during pulmonary vein isolation in patients with atrial fibrillation: impact on outcome and complications. AB - BACKGROUND: The objective of this study was to assess the impact of intracardiac echocardiography (ICE) on the long-term success and complications in patients undergoing pulmonary vein isolation (PVI) for treatment of atrial fibrillation (AF). METHODS AND RESULTS: Three hundred fifteen patients underwent PVI for treatment of AF. Each patient underwent ostial isolation of all PVs using a cooled-tip ablation catheter. PVI was performed using circular mapping (CM) alone (group 1, 56 patients), CM and ICE (group 2, 107 patients), and CM and ICE with titration of radiofrequency energy based on visualization of microbubbles by ICE (group 3, 152 patients). After a mean follow-up time of 417+/-145 days, 19.6% (11 of 56), 16.8% (18 of 107), and 9.8% (15 of 152) of patients in groups 1, 2, and 3 experienced recurrence of AF, respectively. Moreover, whereas no group 3 patient experienced severe (>70%) PV stenosis, severe PV stenosis was documented in 3 (3.5%) of 56 patients in group 1 and in 2 (1.8%) of 107 patients in group 2 (P<0.05). No embolic events were detected in group 3 patients. CONCLUSIONS: Intracardiac echocardiography improves the outcome of cooled-tip PVI. Power adjustment guided by direct visualization of microbubble formation reduces the risk of PV stenosis and improves long-term cure. PMID- 12756154 TI - Mulibrey heart disease: clinical manifestations, long-term course, and results of pericardiectomy in a series of 49 patients born before 1985. AB - BACKGROUND: Mulibrey nanism is a rare inherited disease characterized by growth failure and multiorgan manifestations, including constrictive pericarditis. Its long-term course, the results of pericardiectomy, and the details of myocardial involvement have not been reported previously. METHODS AND RESULTS: We studied 49 patients (26 men) born before 1985 and followed for up to 25 years. By 1999, 25 patients (51%) had developed congestive heart failure (CHF), 19 (39%) had undergone pericardiectomy for constrictive pericarditis, 10 (22%) had died of cardiac causes, and 5 (10%) had died of noncardiac causes. Of the 19 pericardiectomized patients, 12 derived lasting clinical benefit, whereas 1 patient suffered an early noncardiac death and 6 died later of unrelieved or recurrent CHF. At echocardiography in 34 living patients, left ventricular mass adjusted for body height and weight averaged (+/-SEM) 149+/-5 g in 21 unoperated patients, 144+/-8 g in 13 pericardiectomized patients, and 104+/-7 g in 16 healthy persons matched for age and sex (P=0.000). Autopsies of 11 patients showed fibrotic thickening of the pericardial leaves with myocardial hypertrophy and variable but mostly mild myocardial fibrosis. Endocardial thickening was seen in 3 patients. CONCLUSIONS: Constrictive pericarditis, myocardial hypertrophy, and variable myocardial fibrosis constitute the main elements of Mulibrey heart disease. At least one half of patients ultimately develop CHF. Pericardiectomy generally provides clinical benefit, but in approximately one third of patients, CHF may recur because of coexisting myocardial involvement. PMID- 12756155 TI - Temporal trends in sudden cardiac arrest: a 25-year emergency medical services perspective. AB - BACKGROUND: Little is known about temporal trends in survival and prognostic characteristics of patients with out-of-hospital cardiac arrest treated by emergency medical services (EMS). We hypothesized that an evolving combination of beneficial and adverse factors may contribute to temporal patterns of survival. METHODS AND RESULTS: We evaluated a population-based cohort of EMS-treated adult patients with cardiac arrest (n=12 591) from 1977 to 2001 in King County, Washington. Time was grouped into an initial 5-year period and 5 successive 4 year periods. We sought to determine the potential impact of temporal changes in prognostic factors typically beyond EMS control termed "fate" factors (for example, patient age) and factors implemented by EMS termed "program" factors (programs of dispatcher-assisted cardiopulmonary resuscitation and basic life support defibrillation). Several characteristics associated with survival changed over time. Observed survival did not change over time among all patients with cardiac arrest (OR=0.98 [0.95, 1.01], trend for each successive time period) and improved over time among patients with witnessed ventricular fibrillation (OR=1.05 [1.01, 1.09]). In models that included all patients with cardiac arrest and controlled for fate factors, advancing time period was associated with an increase in survival (OR=1.08 [1.05, 1.11]). Conversely, in models that controlled for program factors, advancing time period was associated with a decrease in survival (OR=0.95 [0.93, 0.98]). Results were similar among patients with witnessed ventricular fibrillation. CONCLUSIONS: The static temporal pattern of survival from cardiac arrest appeared to result from an evolving balance of prognostic factors. Programs implemented by EMS appeared to counter adverse temporal trends in prognostic factors typically beyond EMS control. PMID- 12756156 TI - Microvascular filtration is increased in postural tachycardia syndrome. AB - BACKGROUND: Postural tachycardia syndrome (POTS) is related to defective peripheral vasoconstriction of dependent extremities with redistributive hypovolemia. METHODS AND RESULTS: To test whether enhanced microvascular filtration produces leg enlargement, we studied 12 patients 13 to 19 years of age with POTS and defective leg vasoconstriction and 13 age-matched healthy control subjects, with strain-gauge plethysmography used to measure venous pressure (Pv), forearm and calf blood flow, vascular capacitance, and the microvascular filtration coefficient (Kf). Measurements were made while the patient was supine and at steady state during upright tilt to 35 degrees. Supine Pv was not different in POTS, but upright leg Pv tended to be increased above control. Arm and leg peripheral arterial resistance was decreased in the supine and upright positions in patients with POTS compared with control subjects (P=0.01, upright legs). Supine Kf was not significantly increased in the forearm in patients with POTS but was increased in the calf (9.3+/-2.2 versus 5.7+/-2.4 [10(-3)] mL/100 mL per minute per mm Hg, P=0.04), correlating with calf blood flow (rs=0.84, P=0.002). Kf was invariant with orthostasis. The hydraulic contribution to upright filtered flow at 35 degrees tilt, the product of Kf and Pv, was approximately twice that of control (0.41+/-0.09 versus 0.19+/-0.04 mL/100 mL per minute, P=0.04). CONCLUSIONS: Increased microvascular filtration accounts for enhanced leg swelling in patients with POTS with increased arterial blood flow. PMID- 12756157 TI - Improving guideline adherence: a randomized trial evaluating strategies to increase beta-blocker use in heart failure. AB - BACKGROUND: The dissemination of clinical practice guidelines often has not been accompanied by desired improvements in guideline adherence. This study evaluated interventions for implementing a new practice guideline advocating the use of beta-blockers for heart failure patients. METHODS AND RESULTS: This was a randomized controlled trial involving heart failure patients (n=169) with an ejection fraction < or =45% and no contraindications to beta-blockers. Patients' primary providers were randomized in a stratified design to 1 of 3 interventions: (1) control: provider education; (2) provider and patient notification: computerized provider reminders and patient letters advocating beta-blockers; and (3) nurse facilitator: supervised nurse to initiate and titrate beta-blockers. The primary outcome, the proportion of patients who were initiated or uptitrated and maintained on beta-blockers, analyzed by intention to treat, was achieved in 67% (36 of 54) of patients in the nurse facilitator group compared with 16% (10 of 64) in the provider/patient notification and 27% (14 of 51) in the control groups (P<0.001 for the comparisons between the nurse facilitator group and both other groups). The proportion of patients on target beta-blocker doses at the study end (median follow-up, 12 months) was also highest in the nurse facilitator group (43%) compared with the control (10%) and provider/patient notification groups (2%) (P<0.001). There were no differences in adverse events among groups. CONCLUSIONS: The use of a nurse facilitator was a successful approach for implementing a beta-blocker guideline in heart failure patients. The use of provider education, clinical reminders, and patient education was of limited value in this setting. PMID- 12756158 TI - Low paraoxonase activity predicts coronary events in the Caerphilly Prospective Study. AB - BACKGROUND: The hypothesis that paraoxonase (PON1) has a role in preventing atherosclerosis is based on experimental, transgenic, and case-control studies but has not previously been tested prospectively. METHODS AND RESULTS: The Caerphilly Prospective Study is a cohort study of men aged 49 to 65 years observed for coronary heart disease (CHD) events (fatal and nonfatal myocardial infarction) over a mean period of 15 years. Serum PON1 activity toward paraoxon was measured in 1353 participants. PON1 activity was 20% lower in the 163 men who had a coronary event (P=0.039). Men in the highest quintile of PON1 activity had a decreased risk compared with those in the lowest quintile (OR 0.57 [95% CI, 0.33 to 0.96]). The inverse relationship between quintiles of serum PON1 activity and CHD risk was graded, the median change in OR across each quintile being 0.87 (0.77 to 0.98). After adjustment for all other CHD risk factors, including HDL cholesterol, this median value became 0.90 (0.78 to 1.02). PON1 was most predictive of a new CHD event in patients at highest risk by virtue of preexisting CHD (adjusted median OR for each quintile, 0.74 [0.59 to 0.93]; n=313) or the presence of other risk factors. For the highest tertile of CHD risk (n=390) calculated by the Framingham equation, adjusted median OR for each quintile was 0.84 (0.66 to 1.05); n=390. CONCLUSIONS: Low serum PON1 activity toward paraoxon is an independent risk factor for coronary events in men at high risk because of preexisting disease or other CHD risk factors. PMID- 12756159 TI - Combined steroid treatment for congenital heart surgery improves oxygen delivery and reduces postbypass inflammatory mediator expression. AB - BACKGROUND: Steroid administration during cardiopulmonary bypass is thought to improve cardiopulmonary function by modulating bypass-related inflammation. This study was designed to compare preoperative and intraoperative methylprednisolone (MP) to intraoperative MP alone with respect to postbypass inflammation and clinical outcome. METHODS AND RESULTS: Twenty-nine pediatric patients undergoing bypass procedures were randomly assigned to receive preoperative and intraoperative MP (30 mg/kg 4 hours before bypass and in bypass prime, n=14) or intraoperative MP only (30 mg/kg, n=15). Myocardial inflammatory mediator mRNA expression was determined in paired atrial biopsies (before and after bypass) by ribonuclease protection. Before and after bypass, serum IL-6 and IL-10 were measured by ELISA. Postoperative outcome was assessed by intubation time, CICU length of stay, fluid balance, arterio-venous O2 difference (DeltaA-VO2), and inotrope requirements. Compared with intraoperative MP alone, combined preoperative and intraoperative MP was associated with reduced myocardial mRNA expression for IL-6, MCP-1, and ICAM-1 both before and after bypass (P<0.05). Patients who received combined steroids had lower serum IL-6 and increased IL-10 at end-bypass (P<0.05), although differences were negligible by 24 hours. Combined MP treatment was associated with reduced fluid requirements, lower body temperature, and lower DeltaA-VO2 for the first 24 hours after surgery (P<0.05), along with trends toward improvement in other clinical outcomes. CONCLUSIONS: Compared with intraoperative steroid treatment, combined preoperative and intraoperative steroid administration attenuates inflammatory mediator expression more effectively and is associated with improved indexes of O2 delivery in the first 24 hours after congenital heart surgery. These findings need to be confirmed in a larger multicenter trial. PMID- 12756160 TI - Real-time magnetic resonance imaging-guided coronary catheterization in swine. AB - BACKGROUND: We tested the hypothesis that real-time magnetic resonance imaging (MRI) can guide coronary artery catheterization in swine via a percutaneous femoral artery approach. METHODS AND RESULTS: In 12 pigs, we accessed femoral arteries percutaneously. We used 6- or 7-French coronary Judkins catheters filled with dilute 4% gadolinium (Gd) contrast agent and coaxially inserted 0.030-inch diameter active guidewires as endovascular devices. For catheter tracking, we used a 2-dimensional (2D) inversion recovery-prepared spoiled gradient echo sequence at a temporal resolution of 7 frames/s. For guidewire tracking, we used 2D steady-state free precession imaging at a temporal resolution of 9 frames/s. Coronary artery catheterization under MRI guidance was successful in 12/12 pigs. Successful coronary catheterization was verified by obtaining MR angiographic images after direct catheter-based injections of dilute Gd. CONCLUSIONS: Real time MRI-guided catheterization of coronary arteries in swine is feasible via a percutaneous femoral artery approach. Selective coronary MR angiography can then be performed with dilute contrast agent injections. PMID- 12756161 TI - Risks of using internal thoracic artery grafts in patients in chronic hemodialysis via upper extremity arteriovenous fistula. AB - BACKGROUND: In patients in chronic hemodialysis via upper extremity arteriovenous fistula in whom ipsilateral internal thoracic artery graft was used for myocardial revascularization, hemodynamic interference between the fistula and the graft during dialysis can be hypothesized. METHODS AND RESULTS: In 5 patients undergoing chronic hemodialysis via upper extremity arteriovenous fistula, ipsilateral to an internal thoracic to left anterior descending graft mammary flow was studied by means of transthoracic echo-color Doppler at baseline and during hemodialysis. Flow in the contralateral mammary artery was used as control. Transthoracic echocardiography was performed in concomitance with flow evaluation to assess eventual modifications of left ventricular segmental wall motion. Immediately after hemodialysis pump start there was a marked reduction of peak systolic and end-diastolic velocities and time average mean velocity and flow in the ITA ipsilateral to the fistula, whereas no substantial hemodynamic modification was evident in the contralateral artery. Dialysis-induced reduction of ipsilateral ITA flow was accompanied by evidence of hypokinesia of the anterior left ventricular wall. Three cases also experienced clinical angina. CONCLUSIONS: Hemodynamically evident flow steal and consequent myocardial ischemia develop during hemodialysis in patients with upper extremity arteriovenous fistula and ipsilateral internal thoracic artery to coronary graft. These data have major implications for patients' management, both for nephrologists and cardiac surgeons. PMID- 12756163 TI - Prenatal origin of childhood acute myeloid leukemias harboring chromosomal rearrangements t(15;17) and inv(16). PMID- 12756162 TI - Fcgamma receptor transmembrane domains: role in cell surface expression, gamma chain interaction, and phagocytosis. AB - We constructed chimeric receptors to dissect the role of the transmembrane (TM) domain in cell surface expression of and phagocytosis by the gamma chain dependent Fcgamma receptors FcgammaRIIIA and FcgammaRI. FcgammaR chimeras containing the TM and cytoplasmic (CY) domains of the gamma chain were expressed on the cell surface and mediated an efficient phagocytic signal. In contrast, chimeras containing the FcgammaRIIIA TM were poorly expressed. Receptors containing the FcgammaRI TM and the gamma chain CY but lacking the gamma chain TM also were expressed efficiently and mediated phagocytosis, suggesting that a gamma chain dimer induced by the gamma chain TM is not required for efficient phagocytosis. Cotransfection of FcgammaRI or FcgammaRIIIA with the chimera CD8 gamma-gamma (EC-TM-CY) resulted in FcgammaR cell surface expression and phagocytosis, whereas CD8-CD8-gamma, whose TM does not associate with FcgammaR, allowed cell surface expression of (but not phagocytosis by) FcgammaRI. CD8-CD8 gamma also did not allow surface expression of FcgammaRIIIA. Exchanging FcgammaRI and CD8 TMs indicated that the C-terminal 11 amino acids of the FcgammaRI TM are essential for association of FcgammaRI with the gamma chain and phagocytosis. The data indicate that specific sequences in the FcgammaRIIIA and FcgammaRI TMs govern their different interactions with the gamma chain in cell surface expression and phagocytosis and that gamma chain TM sequences are not required for gamma chain-mediated phagocytosis. The data identify a specific region of the FcgammaRI TM and its asparagine as important for FcgammaRI cell surface expression in the absence of the gamma chain and for distinguishing the FcgammaRI and FcgammaRIIIA phenotypes. PMID- 12756164 TI - Imatinib normalizes bone marrow vascularity in patients with chronic myeloid leukemia in first chronic phase. PMID- 12756165 TI - A monoclonal melanoma-specific T-cell population phenotypically indistinguishable from CD3+ LGL-leukemia. PMID- 12756166 TI - 5-Aza-2'-deoxycytidine (decitabine) treatment of hematopoietic malignancies: a multimechanism therapeutic approach? PMID- 12756167 TI - Scabrous and Gp150 are endosomal proteins that regulate Notch activity. AB - Notch and Delta are required for lateral inhibition during eye development. They prevent a tenfold excess in R8 photoreceptor cell specification. Mutations in two other genes, Scabrous and Gp150, result in more modestly increased R8 specification. Their roles in Notch signaling have been unclear. Both sca and gp150 are required for ectopic Notch activity that occurs in the split mutant. Similar phenotypes showed that sca and gp150 genes act in a common pathway. Gp150 was required for all activities of Sca, including inhibition of Notch activity and association with Notch-expressing cells that occur when Sca is ectopically expressed. Mosaic analysis found that the gp150 and sca genes were required in different cells from one another. Gp150 concentrated Sca protein in late endosomes. A model is proposed in which endosomal Sca and Gp150 promote Notch activation in response to Delta, by regulating acquisition of insensitivity to Delta in a subset of cells. PMID- 12756168 TI - Notch activity in neural cells triggered by a mutant allele with altered glycosylation. AB - The receptor protein Notch is inactive in neural precursor cells despite neighboring cells expressing ligands. We investigated specification of the R8 neural photoreceptor cells that initiate differentiation of each Drosophila ommatidium. The ligand Delta was required in R8 cells themselves, consistent with a lateral inhibitor function for Delta. By contrast, Delta expressed in cells adjacent to R8 could not activate Notch in R8 cells. The split mutation of Notch was found to activate signaling in R8 precursor cells, blocking differentiation and leading to altered development and neural cell death. split did not affect other, inductive functions of Notch. The Ile578-->Thr578 substitution responsible for the split mutation introduced a new site for O-fucosylation on EGF repeat 14 of the Notch extracellular domain. The O-fucose monosaccharide did not require extension by Fringe to confer the phenotype. Our results suggest functional differences between Notch in neural and non-neural cells. R8 precursor cells are protected from lateral inhibition by Delta. The protection is affected by modifications of a particular EGF repeat in the Notch extracellular domain. These results suggest that the pattern of neurogenesis is determined by blocking Notch signaling, as well as by activating Notch signaling. PMID- 12756169 TI - Drosophila TGIF is essential for developmentally regulated transcription in spermatogenesis. AB - We have investigated the role of TGIF, a TALE-class homeodomain transcription factor, in Drosophila development. In vertebrates, TGIF has been implicated, by in vitro analysis, in several pathways, most notably as a repressor modulating the response to TGFbeta signalling. Human TGIF has been associated with the developmental disorder holoprosencephaly. Drosophila TGIF is represented by the products of two tandemly repeated highly similar genes, achintya and vismay. We have generated mutations that delete both genes. Homozygous mutant flies are viable and appear morphologically normal, but the males are completely sterile. The defect lies at the primary spermatocyte stage and differentiation is blocked prior to the onset of the meiotic divisions. We show that mutants lacking TGIF function fail to activate transcription of many genes required for sperm manufacture and of some genes required for entry into the meiotic divisions. This groups TGIF together with two other genes producing similar phenotypes, always early and cookie monster, as components of the machinery required for the activation of the spermatogenic programme of transcription. TGIF is the first sequence-specific transcription factor identified in this pathway. By immunolabelling in mouse testes we show that TGIF is expressed in the early stages of spermatogenesis consistent with a conserved role in the activation of the spermatogenesis transcription programme. PMID- 12756170 TI - Requirement for two nearly identical TGIF-related homeobox genes in Drosophila spermatogenesis. AB - The genetic analysis of spermatogenesis in Drosophila melanogaster has led to the identification of several genes that control the onset of meiosis, spermatid differentiation, or both. We described two tightly linked and nearly identical homeobox genes of the TGIF (TG-interacting factor) subclass called vismay and achintya that are essential for spermatogenesis in Drosophila. In flies deficient for both genes, spermatogenesis is blocked prior to any spermatid differentiation and before the first meiotic division. This suggests that vismay and achintya function at the same step as two previously characterized meiotic arrest genes, always early and cookie monster. Consistent with this idea, both always early and cookie monster are still expressed in flies deficient in vismay and achintya. Conversely, Vismay and Achintya proteins are present in always early mutant testes. Co-immunoprecipitation experiments further suggest that Vismay and Achintya proteins exist in a complex with Always early and Cookie monster proteins. Because Vismay and Achintya are likely to be sequence-specific DNA binding factors, these results suggest that they help to specify the spermatogenesis program by recruiting or stabilizing Always early and Cookie monster to specific target genes that need to be transcriptionally regulated during testes development. PMID- 12756171 TI - Endoplasmic reticulum reorganizations and Ca2+ signaling in maturing and fertilized oocytes of marine protostome worms: the roles of MAPKs and MPF. AB - Before a proper Ca(2+) response is produced at fertilization, oocytes typically undergo a maturation process during which their endoplasmic reticulum (ER) is restructured. In marine protostome worms belonging to the phylum Nemertea, the ER of maturing oocytes forms numerous distinct clusters that are about 5 micro m in diameter. After fertilization, mature oocytes with such aggregates generate a normal series of Ca(2+) oscillations and eventually disassemble their ER clusters at around the time that the oscillations cease. Immature oocytes, however, lack prominent ER clusters and fail to exhibit repetitive Ca(2+) oscillations upon insemination, collectively suggesting that cell cycle-related changes in ER structure may play a role in Ca(2+) signaling. To assess the effects of meiotic regulators on the morphology of the ER and the type of Ca(2+) response that is produced at fertilization, nemertean oocytes were treated with pharmacological modulators of mitogen-activated protein kinases (MAPKs) or maturation-promoting factor (MPF) prior to confocal microscopic analyses. Based on such imaging studies and correlative assays of kinase activities, MAPKs of the ERK1/2 type (extracellular signal regulated kinases 1/2) do not seem to be essential for either structural reorganizations of the ER or repetitive Ca(2+) signaling at fertilization. Conversely, MPF levels appear to modulate both ER structure and the capacity to produce normal Ca(2+) oscillations. The significance of these findings is discussed with respect to other reports on ER structure, MPF cycling and Ca(2+) signaling in oocytes of deuterostome animals. PMID- 12756172 TI - The C. elegans Hand gene controls embryogenesis and early gonadogenesis. AB - The C. elegans genome encodes a single Hand bHLH transcription factor. Either hnd 1(RNAi) or a hnd-1 deletion causes partially penetrant defects in viability and gonadogenesis. Dead embryos and young larvae are often misshapen at the posterior end. Our primary focus has been the role of hnd-1 in gonadogenesis. Wild-type C. elegans has two somatic gonadal precursors and two primordial germ cells in stereotyped positions within its four-celled gonadal primordium. The hnd-1 gene affects the presence and position of both the somatic gonadal precursors and primordial germ cells within the primordium, but does not appear to have any role in later gonadogenesis. hnd-1 probably acts within the somatic gonadal precursors or their mesodermal predecessors; defects in primordial germ cells and germ line appear to be secondary. In hnd-1 mutants, somatic gonadal precursors are generated normally, but are not maintained properly and sometimes die. A similar role in controlling the maintenance of precursor fates has been described for other genes governing early organogenesis, including the zebrafish Hand gene hands off. We also report the discovery of two genes, ehn-1 and ehn-3, that have overlapping functions with hnd-1 in embryogenesis and gonadogenesis. PMID- 12756173 TI - Epidermal patterning genes are active during embryogenesis in Arabidopsis. AB - Epidermal cells in the root of Arabidopsis seedling differentiate either as hair or non-hair cells, while in the hypocotyl they become either stomatal or elongated cells. WEREWOLF (WER) and GLABRA2 (GL2) are positive regulators of non hair and elongated cell development. CAPRICE (CPC) is a positive regulator of hair cell development in the root. We show that WER, GL2 and CPC are expressed and active during the stages of embryogenesis when the pattern of cells in the epidermis of the root-hypocotyl axis forms. GL2 is first expressed in the future epidermis in the heart stage embryo and its expression is progressively restricted to those cells that will acquire a non-hair identity in the transition between torpedo and mature stage. The expression of GL2 at the heart stage requires WER function. WER and CPC are transiently expressed throughout the root epidermal layer in the torpedo stage embryo when the cell-specific pattern of GL2 expression is being established in the epidermis. We also show that WER positively regulates CPC transcription and GL2 negatively regulates WER transcription in the mature embryo. We propose that the restriction of GL2 to the future non-hair cells in the root epidermis can be correlated with the activities of WER and CPC during torpedo stage. In the embryonic hypocotyl we show that WER controls GL2 expression. We also provide evidence indicating that CPC may also regulate GL2 expression in the hypocotyl. PMID- 12756174 TI - Retinal pigmented epithelium determination requires the redundant activities of Pax2 and Pax6. AB - The transcription factors Pax2 and Pax6 are co-expressed in the entire optic vesicle (OV) prior and concomitant with the establishment of distinct neuroretinal, retinal, pigmented-epithelial and optic-stalk progenitor domains, suggesting redundant functions during retinal determination. Pax2; Pax6 compound mutants display a dose-dependent reduction in the expression of the melanocyte determinant Mitf, accompanied by transdifferentiation of retinal pigmented epithelium (RPE) into neuroretina (NR) in Pax2(-/-); Pax6(+/-) embryos, which strongly resembles the phenotype of Mitf-null mutants. In Pax2(-/-); Pax6(-/-) OVs Mitf fails to be expressed and NR markers occupy the area that usually represents the Mitf(+) RPE domain. Furthermore, both, Pax2 and Pax6 bind to and activate a MITF RPE-promoter element in vitro, whereas prolonged expression of Pax6 in the Pax2-positive optic stalk leads to ectopic Mitf expression and RPE differentiation in vivo. Together, these results demonstrate that the redundant activities of Pax2 and Pax6 direct the determination of RPE, potentially by directly controlling the expression of RPE determinants. PMID- 12756175 TI - Alx1, a member of the Cart1/Alx3/Alx4 subfamily of Paired-class homeodomain proteins, is an essential component of the gene network controlling skeletogenic fate specification in the sea urchin embryo. AB - In the sea urchin embryo, the large micromeres and their progeny function as a critical signaling center and execute a complex morphogenetic program. We have identified a new and essential component of the gene network that controls large micromere specification, the homeodomain protein Alx1. Alx1 is expressed exclusively by cells of the large micromere lineage beginning in the first interphase after the large micromeres are born. Morpholino studies demonstrate that Alx1 is essential at an early stage of specification and controls downstream genes required for epithelial-mesenchymal transition and biomineralization. Expression of Alx1 is cell autonomous and regulated maternally through beta catenin and its downstream effector, Pmar1. Alx1 expression can be activated in other cell lineages at much later stages of development, however, through a regulative pathway of skeletogenesis that is responsive to cell signaling. The Alx1 protein is highly conserved among euechinoid sea urchins and is closely related to the Cart1/Alx3/Alx4 family of vertebrate homeodomain proteins. In vertebrates, these proteins regulate the formation of skeletal elements of the limbs, face and neck. Our findings suggest that the ancestral deuterostome had a population of biomineral-forming mesenchyme cells that expressed an Alx1-like protein. PMID- 12756176 TI - The glial sling is a migratory population of developing neurons. AB - For two decades the glial sling has been hypothesized to act as a guidance substratum for developing callosal axons. However, neither the cellular nature of the sling nor its guidance properties have ever been clearly identified. Although originally thought to be glioblasts, we show here that the subventricular zone cells forming the sling are in fact neurons. Sling cells label with a number of neuronal markers and display electrophysiological properties characteristic of neurons and not glia. Furthermore, sling cells are continuously generated until early postnatal stages and do not appear to undergo widespread cell death. These data indicate that the sling may be a source of, or migratory pathway for, developing neurons in the rostral forebrain, suggesting additional functions for the sling independent of callosal axon guidance. PMID- 12756177 TI - Two Pax genes, eye gone and eyeless, act cooperatively in promoting Drosophila eye development. AB - We report the identification of a Drosophila Pax gene, eye gone (eyg), which is required for eye development. Loss-of-function eyg mutations cause reduction or absence of the eye. Similar to the Pax6 eyeless (ey) gene, ectopic expression of eyg induces extra eye formation, but at sites different from those induced by ey. Several lines of evidence suggest that eyg and ey act cooperatively: (1) eyg expression is not regulated by ey, nor does it regulate ey expression, (2) eyg induced ectopic morphogenetic furrow formation does not require ey, nor does ey induced ectopic eye production require eyg, (3) eyg and ey can partially substitute for the function of the other, and (4) coexpression of eyg and ey has a synergistic enhancement of ectopic eye formation. Our results also show that eyg has two major functions: to promote cell proliferation in the eye disc and to promote eye development through suppression of wg transcription. PMID- 12756178 TI - TagA, a putative serine protease/ABC transporter of Dictyostelium that is required for cell fate determination at the onset of development. AB - The tag genes of Dictyostelium are predicted to encode multi-domain proteins consisting of serine protease and ATP-binding cassette transporter domains. We have identified a novel tag gene, tagA, which is involved in cell type differentiation. The tagA mRNA accumulates during the first four hours of development, whereas TagA protein accumulates between two and ten hours of development and decreases thereafter. Wild-type cells express tagA in prespore cells and mature spores, defining tagA expression as prespore specific. However, tagA mutant cells that activate the tagA promoter do not sporulate, but instead form part of the outer basal disc and lower cup of the fruiting body. tagA mutant aggregates elaborate multiple prestalk cell regions during development and produce spores asynchronously and with low viability. tagA mutants produce about twice as many prestalk cells as the wild type as judged by a prestalk cell reporter construct. When mixed with wild-type cells, tagA(-) cells become overrepresented in the prestalk cell population, suggesting that this phenotype is cell-autonomous. These results suggest that TagA is required for the specification of an initial population of prespore cells in which tagA is expressed. Expression profiling uncovered a delay in the transcriptional program between 2 and 6 hours, coincident with TagA expression, revealing an early function for TagA. TagA also appears to play a general role in cell fate determination since tagA mutants express a spore coat protein gene (cotB) within vacuolated cells that form part of the stalk and they express a prestalk/stalk specific gene (ecmB) within cells that become spores. The expression of TagA at two hours of development, the observed coincident delay in the transcriptional program and the subsequent mis-expression of cell-type specific genes provide evidence for cell fate determination beginning in some cells much earlier than previously believed. PMID- 12756180 TI - Somatic motoneurone specification in the hindbrain: the influence of somite derived signals, retinoic acid and Hoxa3. AB - We have investigated the mechanisms involved in generating hindbrain motoneurone subtypes, focusing on somatic motoneurones, which are confined to the caudal hindbrain within rhombomeres 5-8. Following heterotopic transplantation of rhombomeres along the rostrocaudal axis at various developmental stages, we have found that the capacity of rhombomeres to generate somatic motoneurones is labile at the neural plate stage but becomes fixed just after neural tube closure, at stage 10-11. Grafting of somites or retinoic acid-loaded beads beneath the rostral hindbrain induced the formation of somatic motoneurones in rhombomere 4 only, and Hox genes normally expressed more caudally (Hoxa3, Hoxd4) were induced in this region. Targeted overexpression of Hoxa3 in the rostral hindbrain led to the generation of ectopic somatic motoneurones in ventral rhombomeres 1-4, and was accompanied by the repression of the dorsoventral patterning gene Irx3. Taken together, these observations suggest that the somites, retinoic acid and Hox genes play a role in patterning somatic motoneurones in vivo. PMID- 12756179 TI - Retinal ganglion cell-derived sonic hedgehog signaling is required for optic disc and stalk neuroepithelial cell development. AB - The development of optic stalk neuroepithelial cells depends on Hedgehog (Hh) signaling, yet the source(s) of Hh protein in the optic stalk is unknown. We provide genetic evidence that sonic hedgehog (Shh) from retinal ganglion cells (RGCs) promotes the development of optic disc and stalk neuroepithelial cells. We demonstrate that RGCs express Shh soon after differentiation, and cells at the optic disc in close proximity to the Shh-expressing RGCs upregulate Hh target genes, which suggests they are responding to RGC-derived Shh signaling. Conditional ablation of Shh in RGCs caused a complete loss of optic disc astrocyte precursor cells, resulting in defective axon guidance in the retina, as well as conversion of the neuroepithelial cells in the optic stalk to pigmented cells. We further show that Shh signaling modulates the size of the Pax2(+) astrocyte precursor cell population at the optic disc in vitro. Together, these data provide a novel insight into the source of Hh that promotes neuroepithelial cell development in the mammalian optic disc and stalk. PMID- 12756181 TI - giant nuclei is essential in the cell cycle transition from meiosis to mitosis. AB - At the transition from meiosis to cleavage mitoses, Drosophila requires the cell cycle regulators encoded by the genes, giant nuclei (gnu), plutonium (plu) and pan gu (png). Embryos lacking Gnu protein undergo DNA replication and centrosome proliferation without chromosome condensation or mitotic segregation. We have identified the gnu gene encoding a novel phosphoprotein dephosphorylated by Protein phosphatase 1 at egg activation. Gnu is normally expressed in the nurse cells and oocyte of the ovary and is degraded during the embryonic cleavage mitoses. Ovarian death and sterility result from gnu gain of function. gnu function requires the activity of pan gu and plu. PMID- 12756182 TI - Strabismus is asymmetrically localised and binds to Prickle and Dishevelled during Drosophila planar polarity patterning. AB - Planar polarity decisions in the wing of Drosophila involve the assembly of asymmetric protein complexes containing the conserved receptor Frizzled. In this study, we analyse the role of the Van Gogh/strabismus gene in the formation of these complexes and cell polarisation. We find that the Strabismus protein becomes asymmetrically localised to the proximal edge of cells. In the absence of strabismus activity, the planar polarity proteins Dishevelled and Prickle are mislocalised in the cell. We show that Strabismus binds directly to Dishevelled and Prickle and is able to recruit them to membranes. Furthermore, we demonstrate that the putative PDZ-binding motif at the C terminus of Strabismus is not required for its function. We propose a two-step model for assembly of Frizzledcontaining asymmetric protein complexes at cell boundaries. First, Strabismus acts together with Frizzled and the atypical cadherin Flamingo to mediate apicolateral recruitment of planar polarity proteins including Dishevelled and Prickle. In the second phase, Dishevelled and Prickle are required for these proteins to become asymmetrically distributed on the proximodistal axis. PMID- 12756183 TI - pannier and pointedP2 act sequentially to regulate Drosophila heart development. AB - The Drosophila heart consists of two major cell types: cardioblasts, which form the contractile tube of the heart; and pericardial cells, which flank the cardioblasts and are thought to filter and detoxify the blood or hemolymph of the fly. We present the completion of the entire cell lineage of all heart cells. Notably, we detect a previously unappreciated distinction between the lineages of heart cells located in the posterior seven segments relative to those located more anteriorly. Using a genetic screen, we have identified the ETS-transcription factor pointed as a key regulator of cardioblast and pericardial cell fates in the posterior seven segments of the heart. In this domain, pointed promotes pericardial cell development and opposes cardioblast development. We find that this function of pointed is carried out primarily if not exclusively by the pointedP2 isoform and, that in this context, pointedP2 may act independently of Ras/MAPK pathway activity. We go on to show that the GATA transcription factor pannier acts early in dorsal mesoderm development to promote the development of the cardiac mesoderm and thus all heart cells. Finally, we demonstrate that pannier acts upstream of pointed in a developmental pathway in which pannier promotes cardiac mesoderm formation, and pointed acts subsequently in this domain to distinguish between cardioblast and pericardial cell fates. PMID- 12756184 TI - Gata factor Pannier is required to establish competence for heart progenitor formation. AB - Inductive signaling is of pivotal importance for developmental patterns to form. In Drosophila, the transfer of TGFbeta (Dpp) and Wnt (Wg) signaling information from the ectoderm to the underlying mesoderm induces cardiac-specific differentiation in the presence of Tinman, a mesoderm-specific homeobox transcription factor. We present evidence that the Gata transcription factor, Pannier, and its binding partner U-shaped, also a zinc-finger protein, cooperate in the process of heart development. Loss-of-function and germ layer-specific rescue experiments suggest that pannier provides an essential function in the mesoderm for initiation of cardiac-specific expression of tinman and for specification of the heart primordium. u-shaped also promotes heart development, but unlike pannier, only by maintaining tinman expression in the cardiogenic region. By contrast, pan-mesodermal overexpression of pannier ectopically expands tinman expression, whereas overexpression of u-shaped inhibits cardiogenesis. Both factors are also required for maintaining dpp expression after germ band retraction in the dorsal ectoderm. Thus, we propose that Pannier mediates as well as maintains the cardiogenic Dpp signal. In support, we find that manipulation of pannier activity in either germ layer affects cardiac specification, suggesting that its function is required in both the mesoderm and the ectoderm. PMID- 12756185 TI - Myogenic cells fates are antagonized by Notch only in asymmetric lineages of the Drosophila heart, with or without cell division. AB - During the formation of the Drosophila heart, a combinatorial network that integrates signaling pathways and tissue-specific transcription factors specifies cardiac progenitors, which then undergo symmetric or asymmetric cell divisions to generate the final population of diversified cardiac cell types. Much has been learned concerning the combinatorial genetic network that initiates cardiogenesis, whereas little is known about how exactly these cardiac progenitors divide and generate the diverse population of cardiac cells. In this study, we examined the cell lineages and cell fate determination in the heart by using various cell cycle modifications. By arresting the cardiac progenitor cell divisions at different developing stages, we determined the exact cell lineages for most cardiac cell types. We found that once cardiac progenitors are specified, they can differentiate without further divisions. Interestingly, the progenitors of asymmetric cell lineages adopt a myocardial cell fate as opposed to a pericardial fate when they are unable to divide. These progenitors adopt a pericardial cell fate, however, when cell division is blocked in numb mutants or in embryos with constitutive Notch activity. These results suggest that a numb/Notch-dependent cell fate decision can take place even in undivided progenitors of asymmetric cell divisions. By contrast, in symmetric lineages, which give rise to a single type of myocardial-only or pericardial-only progeny, repression or constitutive activation of the Notch pathway has no apparent effect on progenitor or progeny fate. Thus, inhibition of Notch activity is crucial for specifying a myogenic cell fate only in asymmetric lineages. In addition, we provide evidence that whether or not Suppressor-of-Hairless can become a transcriptional activator is the key switch for the Numb/Notch activity in determining a myocardial versus pericardial cell fate. PMID- 12756186 TI - Mechanism of hedgehog signaling during Drosophila eye development. AB - Although Hedgehog (Hh) signaling is essential for morphogenesis of the Drosophila eye, its exact link to the network of tissue-specific genes that regulate retinal determination has remained elusive. In this report, we demonstrate that the retinal determination gene eyes absent (eya) is the crucial link between the Hedgehog signaling pathway and photoreceptor differentiation. Specifically, we show that the mechanism by which Hh signaling controls initiation of photoreceptor differentiation is to alleviate repression of eya and decapentaplegic (dpp) expression by the zinc-finger transcription factor Cubitus interruptus (Ci(rep)). Furthermore, our results suggest that stabilized, full length Ci (Ci(act)) plays little or no role in Drosophila eye development. Moreover, while the effects of Hh are primarily concentration dependent in other tissues, hh signaling in the eye acts as a binary switch to initiate retinal morphogenesis by inducing expression of the tissue-specific factor Eya. PMID- 12756187 TI - Conditional inactivation of FGF receptor 2 reveals an essential role for FGF signaling in the regulation of osteoblast function and bone growth. AB - Human craniosynostosis syndromes, resulting from activating or neomorphic mutations in fibroblast growth factor receptor 2 (FGFR2), underscore an essential role for FGFR2 signaling in skeletal development. Embryos harboring homozygous null mutations in FGFR2 die prior to skeletogenesis. To address the role of FGFR2 in normal bone development, a conditional gene deletion approach was adopted. Homologous introduction of cre recombinase into the Dermo1 (Twist2) gene locus resulted in robust expression of CRE in mesenchymal condensations giving rise to both osteoblast and chondrocyte lineages. Inactivation of a floxed Fgfr2 allele with Dermo1-cre resulted in mice with skeletal dwarfism and decreased bone density. Although differentiation of the osteoblast lineage was not disturbed, the proliferation of osteoprogenitors and the anabolic function of mature osteoblasts were severely affected. PMID- 12756188 TI - Active lifestyle and diabetes. PMID- 12756189 TI - How do beta-blockers improve ventricular function in patients with congestive heart failure? PMID- 12756190 TI - Transesophageal echocardiography: clinical indications and applications. PMID- 12756191 TI - Coronary collaterals: an important and underexposed aspect of coronary artery disease. PMID- 12756192 TI - Eplerenone: cardiovascular protection. AB - Data from animal studies and clinical trials indicate that aldosterone causes cardiovascular and renal injury through mineralocorticoid receptor-dependent mechanisms. However, although aldosterone receptor antagonism reduces mortality in patients with congestive heart failure, the progestational and antiandrogenic side effects of the nonspecific aldosterone receptor antagonist, spironolactone, have limited its usefulness in the treatment of hypertension. This review provides an overview of the pharmacology, efficacy, and safety of a new, more selective aldosterone receptor antagonist, eplerenone, in the context of emerging concepts of the role of aldosterone in cardiovascular toxicity. PMID- 12756193 TI - Images in cardiovascular medicine. Myocardial fibrosis in fabry disease demonstrated by multislice computed tomography: comparison with biopsy findings. PMID- 12756194 TI - Images in cardiovascular medicine. Pulmonary vein stenosis after radiofrequency ablation for atrial fibrillation: image findings with multiphasic pulmonary magnetic resonance angiography. PMID- 12756195 TI - New lipid-lowering combo proves successful. PMID- 12756196 TI - Genetic polymorphisms of CYP2C8 in Japanese population. AB - CYP2C8 plays important roles in metabolizing therapeutic drugs and endogenous compounds. Although genetic polymorphisms of CYP2C8 were reported, there is little information on CYP2C8 polymorphisms in the Japanese population. In the present study, we screened for previously described polymorphisms in the coding region of this gene using polymerase chain reaction (PCR)-restriction fragment length polymorphism or allele specific-PCR analyses. Eleven polymorphisms of CYP2C8*2 (I269F), CYP2C8*3 (R139K, K399R), CYP2C8*4 (I264M), CYP2C8*5 (frameshift), T130N, E154D, N193K, K249R, L390S, P404A, and H411L have been comprehensively investigated in at least 200 Japanese individuals. A single subject was heterozygous for CYP2C8*5, and the allele frequency was calculated as 0.0025. The other single nucleotide polymorphisms (SNPs) were not found in the Japanese subjects in the present study. Thus, it appears that the frequencies of these alleles in Japanese are extremely low. In addition, concerning the SNPs of T130N, E154D, N193K, K249R, and H411L, it remains clear that these alleles exist as polymorphisms or represent sequence errors or cloning artifacts. Although several SNPs such as CYP2C8*2, CYP2C8*3, CYP2C8*4, and P404A have been reported to reduce the enzymatic activity, pharmacokinetic abnormalities of drugs metabolized by polymorphic CYP2C8 might be rare in Japanese. PMID- 12756197 TI - Basolateral active uptake of nitrofurantoin in the CIT3 cell culture model of lactation. AB - Nitrofurantoin and other agents are actively transported into human and rodent milk. The purpose of this study was to determine whether nitrofurantoin active transport across mammary epithelia occurs basolaterally or apically, using the CIT3 cell culture model of lactation. The CIT3 model actively transports nitrofurantoin in the basolateral to apical direction. Basolateral to apical permeability [92.9 +/- 6.6 (microl/h)/cm(2)] was differentially decreased by unlabeled nitrofurantoin (250 microM) on the basolateral, apical, or both sides [49.5 +/- 1.8, 57.9 +/- 1.4, or 48.5 +/- 1.6 (microl/h)/cm(2), respectively]. Apical to basolateral permeability [27.6 +/- 1.8 (microl/h)/cm(2)] was increased in the presence of unlabeled nitrofurantoin (250 microM) on the basolateral, apical, or both sides [36.4 +/- 1.5, 39.9 +/- 0.7, 42.4 +/- 1.1 (microl/h)/cm(2), respectively]. These data indicate a basolateral active uptake mechanism for nitrofurantoin, which remains to be identified. This mechanism may influence the exposure of suckling infants to xenobiotics, as well as having potentially toxic effects on the lactating mammary epithelium and possibly altering the nutritional quality of the milk. PMID- 12756198 TI - NMR identification of an S-linked glucuronide of a triazole thione formed in vitro. AB - The metabolism of 3-([3-(2-Chlorophenyl)-4,5-dihydro-5-thioxo-1H-1,2,4-triazol-1 yl]methyl)benzonitrile (AR-C133611XX) was studied in isolated dog hepatocytes. The major metabolite of AR-C133611XX was characterized by high performance liquid chromatography-mass spectrometry and NMR and found to be the product of direct glucuronidation. Evidence from 1H and 13C-NMR chemical shifts and a long-range proton carbon correlation experiment was used to deduce that glucuronidation had taken place on the sulfur atom. Full NMR data on this unusual metabolite is presented. Substitution or replacement of the sulfur atom resulted in a significant decrease in the observed rate of glucuronidation. PMID- 12756199 TI - Enantioselectivity of human hydroxysteroid sulfotransferase ST2A3 with naphthyl-1 ethanols. AB - Hydroxysteroid (alcohol) sulfotransferases catalyze the sulfation of several endogenous steroids and many hydrophobic xenobiotic alcohols. The substrate stereoselectivities of sulfotransferases may be critically important in determining their overall roles in metabolism of drugs, carcinogens, and other xenobiotics. In the present work, stereoselectivity of the human hydroxysteroid sulfotransferase ST2A3 (also variously named as SULT2A1 or human DHEA-ST) was examined through analysis of its catalytic activities with the enantiomers of 1 naphthyl-1-ethanol and 2-naphthyl-1-ethanol. The kcat/Km value for sulfation of the R-(+)-enantiomer of 1-naphthyl-1-ethanol catalyzed by ST2A3 was 3.3 min-1mM 1, whereas the S-(-)-enantiomer was not a substrate for the enzyme. S-(-)-1 naphthyl-1-ethanol did however interact with ST2A3 as an inhibitor of the sulfation of dehydroepiandrosterone. This substrate stereospecificity was not present with the enantiomers of 2-naphthyl-1-ethanol, since both were substrates for the enzyme. Such differences between the sulfation of 1- and 2-naphthyl-1 ethanol are consistent with the importance of steric interactions between the ethanol group and a hydrogen atom at the peri-position (C8) on the naphthyl ring in 1-naphthyl-1-ethanol that combine with the topology of the enzyme's active site to determine stereospecificity. PMID- 12756200 TI - Quinol-based metabolic cycle for estrogens in rat liver microsomes. AB - According to a recently reported metabolic pathway, phenolic A-ring estrogens are metabolized in rat liver microsomes partially to the corresponding quinols by cytochrome P450 isoenzymes. We found that these quinols could, in turn, undergo reduction to regenerate the parent estrogens consumed during the metabolic process. Among the tested endogenous reducing agents, NADH and especially NADPH produced a significant extent of reductive conversion. Enzymes available in rat liver microsomes further catalyzed this reaction with 6.5 +/- 1.5 nmol. min(-1). (mg of protein)(-1) measured as the initial rate of estrone formation at 37 degrees C, whereas the initial rate of second-order reaction for the reduction of E1-quinol by a 10-fold excess of NADPH in a microsome-free buffer solution and under identical incubation conditions was 0.62 +/- 0.03 nmol. min(-1). The quinol route is, therefore, unique among estrogen-metabolizing pathways for its bioreversibility due to the facile regeneration of the phenolic A-ring estrogens consumed in the preceding oxidative process. PMID- 12756201 TI - High pressure liquid chromatography and mass spectrometry characterization of the nephrotoxic biotransformation products of Cisplatin. AB - Previous studies have shown that cisplatin requires metabolic activation to become nephrotoxic. The activation is proposed to be via the metabolism of a glutathione-platinum conjugate to a cysteinyl-glycine-platinum conjugate, which is further processed to a cysteine conjugate. Preincubating cisplatin with glutathione (GSH), cysteinyl-glycine, or N-acetylcysteine (NAC) results in a transient increase in the toxicity of cisplatin toward renal proximal tubular cells. In this study, the preincubation solutions were analyzed by high pressure liquid chromatography (HPLC), atomic absorption spectrometry, and mass spectrometry (MS) to characterize the formation and structure of the platinum conjugates. HPLC analysis of the cisplatin-GSH, cisplatin-cysteinyl-glycine, and cisplatin-NAC preincubation solutions revealed two new platinum-containing peaks in each of the solutions. MS-MS analysis of the peaks revealed a diplatinum- and a monoplatinum conjugate in each of the solutions. Analysis of the composition and toxicity of the solutions with time showed that the transient increase in toxicity correlated with the formation of the monoplatinum conjugate whereas prolonged preincubation decreased toxicity and correlated with the formation of the diplatinum conjugate. The monoplatinum-monoglutathione conjugate is a substrate for gamma-glutamyl transpeptidase, an enzyme that is essential for the nephrotoxicity of cisplatin. The monoplatinum-mono-NAC conjugate can be deacetylated to a cysteine conjugate, which is a substrate for pyroxidol phosphate (PLP)-dependent cysteine S-conjugate beta-lyase. This PLP-dependent enzyme is proposed to catalyze the final step in the metabolic activation of cisplatin. Identification of the structure and toxicity of these conjugates further elucidates the metabolism of cisplatin to a nephrotoxin. PMID- 12756202 TI - Influence of propiverine on hepatic microsomal cytochrome p450 enzymes in male rats. AB - The bladder spasmolytics propiverine was shown to induce hepatic cytochrome P450 (P450) and aminopyrine and aniline oxidation in rats. To characterize the type of enzyme induction and its dose dependence, activities of seven hepatic microsomal P450-dependent monooxygenases were measured in 72 male LEW1A albino rats (body weight 236-295 g) after oral treatment with 0.5, 2, 6, and 60 mg/kg of propiverine hydrochloride for 5 days and compared with the effects of 40 mg/kg beta-naphthoflavone, 10 mg/kg phenobarbital, and 20 mg/kg dexamethasone (each group, n = 8). CYP2B expression was measured by Western blotting. Furthermore, the inhibitory potency of propiverine on P450 enzymes was evaluated in competition assays with three most specific monooxygenases. Results show that Propiverine induced several monooxygenases and CYP2B expression dose dependently. The effects were well comparable with a phenobarbital-type inducer with 60 mg/kg being equipotent to 10 mg/kg phenobarbital. Furthermore, propiverine in low concentrations inhibited pentylresorufin O-dealkylase (for CYP2B) in vitro. In conclusion, propiverine is a phenobarbital-type inducer on hepatic P450 enzymes in rats in doses about 100-times above the therapeutic doses in man. PMID- 12756203 TI - Distinct tissue distribution of metabolites of the novel glutathione-activated thiopurine prodrugs cis-6-(2-acetylvinylthio)purine and trans-6-(2 acetylvinylthio)guanine and 6-thioguanine in the mouse. AB - The compounds cis-6-(2-acetylvinylthio)purine (cis-AVTP) and trans-6-(2 acetylvinylthio)guanine (trans-AVTG) are glutathione-activated prodrugs of 6 mercaptopurine (6-MP) and 6-thioguanine (6-TG), respectively, that have comparable or lower IC50 values in tumor cells than 6-MP and 6-TG. Previously, we showed that cis-AVTP- and trans-AVTG-treated mice exhibited less bone marrow and intestinal toxicity and excreted a lower fraction of the administered dose in urine than did mice treated with equivalent 6-TG doses. To explain these results, the tissue distribution and levels of metabolites of cis-AVTP, trans-AVTG, and 6 TG were examined at 15, 30, and 45 min after i.p. treatment of mice with equimolar doses of these compounds. After prodrug treatment, the thiopurines, the corresponding thiopurine ribosides and nucleotides, thioxanthine (TX), and thiouric acid (TU) were quantitated in plasma, red blood cells, liver, and intestine. Thiopurine and thiopurine riboside and nucleotide area under the curve between 15 and 45 min [AUC(15-45)] values were generally comparable after cis AVTP and trans-AVTG treatments but were lower than those after 6-TG treatment. A higher liver/plasma metabolite ratio was evident after trans-AVTG treatment than after cis-AVTP or 6-TG treatments, which exhibited similar liver/plasma ratios. Treatment with cis-AVTP yielded the highest AUC(15-45) for TX and TU in plasma, liver, and intestine. Prodrug treatment did not change the concentration of reduced or oxidized glutathione in tissue homogenates. Collectively, these results show distinct patterns of metabolites depending upon the compound used and suggest that differences in metabolite levels and composition after cis-AVTP, trans-AVTG, and 6-TG treatments may partially explain the different toxicity and urinary metabolite excretion profiles previously observed among cis-AVTP, trans AVTG, and 6-TG. PMID- 12756204 TI - Covalent binding of 2-phenylpropionyl-S-acyl-CoA thioester to tissue proteins in vitro. AB - In this study, we investigated the possible involvement of acyl-CoA, reactive intermediary metabolites of 2-arylpropionic acids (profens), in protein adduct formation in rat liver homogenate and in human serum albumin (HSA) in buffer. (RS)-[1-14C]-2-Phenylpropionic acid (14C-2-PPA, 1 mM) was incubated with rat liver homogenate (1.5 mg/ml) in the presence of cofactors of acyl-CoA formation (Mg2+, ATP, and CoA). Aliquots of the incubation mixture were analyzed for covalent binding and acyl-CoA formation over a 3-h period. High-performance liquid chromatographic analysis of the products from such incubations showed the presence of 2-phenylpropionyl-S-acyl-CoA (2-PPA-CoA), which was confirmed by coelution with authentic 2-PPA-CoA, as well as by mass spectrometry. In the same incubations, 2-PPA was shown to bind covalently to hepatic proteins in a time- and ATP-dependent fashion. Inhibition of 2-PPA-CoA formation by acyl-CoA synthetase inhibitors, such as palmitic acid, lauric acid, octanoic acid, and ibuprofen, markedly decreased the extent of covalent binding of 2-PPA to hepatic proteins. Results from these in vitro studies strongly suggest that acyl-CoA thioester derivatives are chemically reactive and are able to bind covalently to tissue proteins in vitro, and, therefore, may contribute significantly to covalent adduct formation of profen drugs in vivo. PMID- 12756205 TI - The disposition of voriconazole in mouse, rat, rabbit, guinea pig, dog, and human. AB - Voriconazole is a new triazole antifungal agent with potent, wide-spectrum activity. Its pharmacokinetics and metabolism have been studied in mouse, rat, rabbit, dog, guinea pig, and humans after single and multiple administration by both oral and intravenous routes. Absorption of voriconazole is essentially complete in all species. The elimination of voriconazole is characterized by non linear pharmacokinetics in all species. Consequently, pharmacokinetic parameters are dependent upon dose, and a superproportional increase in area under the curve is seen with increasing dose in rat and dog toxicology studies. Following multiple administration, there is a decrease in systemic exposure. This is most pronounced in mouse and rat, less so in dog, and not observed in guinea pig or rabbit. Repeat-dose toxicology studies in mouse, rat, and dog have demonstrated that induction of cytochrome P450 by voriconazole (autoinduction of metabolism) is responsible for the decreased exposure in these species. Autoinduction of metabolism is not observed in humans, and plasma steady-state concentrations remain constant with time. Voriconazole is extensively metabolized in all species. The major pathways in humans involve fluoropyrimidine N-oxidation, fluoropyrimidine hydroxylation, and methyl hydroxylation. Also, N-oxidation facilitates cleavage of the molecule, resulting in loss of the fluoropyrimidine moiety and subsequent conjugation with glucuronic acid. Major pathways are represented in animal species. The major circulating metabolite in rat, dog, and human is the N-oxide of voriconazole. It is not thought to contribute to efficacy since it is at least 100-fold less potent than voriconazole against fungal pathogens in vitro. PMID- 12756206 TI - Involvement of CYP3A4, CYP2C8, and CYP2D6 in the metabolism of (R)- and (S) methadone in vitro. AB - To clarify the oxidative metabolism of methadone (R)- and (S)-enantiomers, the depletion of parent (R)- and (S)-methadone and the formation of racemic 2 ethylidene-1,5-dimethyl-3,3-diphe-nylpyrolidine were studied using human liver microsomes and recombinant cytochrome P450 enzymes. Based on studies with isoform selective chemical inhibitors and expressed enzymes, CYP3A4 was the predominant enzyme involved in the metabolism of (R)-methadone. However, it has different stereoselectivity toward (R)- and (S)-methadone. In recombinant CYP3A4, the metabolic clearance of (R)-methadone was about 4-fold higher than that of (S) methadone. CYP2C8 is also involved in the metabolism of methadone, but its contribution to the metabolism of (R)-methadone was smaller than that of CYP3A4. But for the metabolism of (S)-methadone, the roles of CYP2C8 and CYP3A4 appeared equal. Although CYP2D6 is involved in the metabolism of (R)- and (S)-methadone, its role was smaller compared with CYP3A4 and CYP2C8. Using clinically relevant concentrations of ketoconazole (1 microM, selective CYP3A4 inhibitor), trimethoprim (100 microM, selective CYP2C8 inhibitor), and paroxetine (5 microM, potent CYP2D6 inhibitor), these inhibitors decreased the hepatic metabolism of (R)-[(S)-]methadone by 69% (47%), 22% (51%), and 41% (77%), respectively. However, inhibition of the metabolism of (R)- and (S)-methadone by paroxetine was due to inhibition not only of CYP2D6, but also CYP3A4 and, to a minor extent, CYP2C8. The present in vitro findings indicated that CYP3A4, CYP2C8, and CYP2D6 are all involved in the stereoselective metabolism of methadone (R)- and (S) enantiomers. These data suggest that coadministration of inhibitors of CYP3A4 and CYP2C8 may produce clinically significant drug-drug interactions with methadone. PMID- 12756207 TI - In vitro metabolism of chloroquine: identification of CYP2C8, CYP3A4, and CYP2D6 as the main isoforms catalyzing N-desethylchloroquine formation. AB - In humans, the antimalarial drug chloroquine (CQ) is metabolized into one major metabolite, N-desethylchloroquine (DCQ). Using human liver microsomes (HLM) and recombinant human cytochrome P450 (P450), we performed studies to identify the P450 isoform(s) involved in the N-desethylation of CQ. In HLM incubated with CQ, only DCQ could be detected. Apparent Km and Vmax values (mean +/- S.D.) for metabolite formation were 444 +/- 121 microM and 617 +/- 128 pmol/min/mg protein, respectively. In microsomes from a panel of 16 human livers phenotyped for 10 different P450 isoforms, DCQ formation was highly correlated with testosterone 6beta-hydroxylation (r = 0.80; p < 0.001), a CYP3A-mediated reaction, and CYP2C8 mediated paclitaxel alpha-hydroxylation (r = 0.82; p < 0.001). CQ N-desethylation was diminished when coincubated with quercetin (20-40% inhibition), ketoconazole, or troleandomycin (20-30% inhibition) and was strongly inhibited (80% inhibition) by a combination of ketoconazole and quercetin, which further corroborates the contribution of CYP2C8 and CYP3As. Of 10 cDNA-expressed human P450s examined, only CYP1A1, CYP2D6, CYP3A4, and CYP2C8 produced DCQ. CYP2C8 and CYP3A4 constituted low-affinity/high-capacity systems, whereas CYP2D6 was associated with higher affinity but a significantly lower capacity. This property may explain the ability of CQ to inhibit CYP2D6-mediated metabolism in vitro and in vivo. At therapeutically relevant concentrations ( approximately 100 microM CQ in the liver), CYP2C8, CYP3A4, and, to a much lesser extent, CYP2D6 are expected to account for most of the CQ N-desethylation. PMID- 12756208 TI - Comparative analysis of CYP3A expression in human liver suggests only a minor role for CYP3A5 in drug metabolism. AB - To study mechanisms behind the interindividual variability in CYP3A expression and the relative contribution of the different CYP3A enzymes to the overall CYP3A activity, we have analyzed CYP3A4, CYP3A5, CYP3A43, and PXR mRNA and CYP3A4 and CYP3A5 protein expression, catalytic activity, and polymorphism in the CYP3A5 gene in a panel of 46 Caucasian human livers. Protein quantification was performed by Western blotting using enzyme-specific antibodies directed to the C termini of CYP3A4 or CYP3A5, and carrier protein-coupled peptides as standards. The mRNA levels were determined by quantitative real-time PCR. CYP3A activity was measured by analysis of the rate of testosterone 6beta-hydroxylation. A correlation existed between all CYP3A and PXR mRNA transcripts measured. The interindividual variability in CYP3A4 and CYP3A5 mRNA expression was higher than that of CYP3A protein and activity. The CYP3A5 protein was expressed at quantifiable levels in 5 (10.9%) of the livers. Four of those were heterozygous for the CYP3A5*1 allele and had CYP3A5 protein at a mean level of 17% of that of total CYP3A, whereas one liver sample was from a CYP3A5*3 homozygote individual having lower amounts of CYP3A5. In total, CYP3A5 only contributed 2% of the overall CYP3A protein among all samples. In conclusion, our data indicate that CYP3A4, CYP3A5, CYP3A43, and PXR hepatic mRNA expression correlate, indicating common regulatory features, and that the contribution of CYP3A5 to hepatic drug metabolism in Caucasians is insignificant. PMID- 12756209 TI - The effect of incubation conditions on the enzyme kinetics of udp glucuronosyltransferases. AB - Traditionally, the Michaelis-Menten equation has been used to determine kinetic parameters for in vitro glucuronidation assays. Recently, estradiol-3-glucuronide formation was shown to exhibit non-Michaelis-Menten kinetics consistent with autoactivation. A concern with the observation of nontraditional kinetics is that they may result as an artifact of the incubation conditions. To examine this concern, the formation of estradiol-3-glucuronide was investigated using human liver microsomes prepared by two different methods, a range of assay conditions, and activation by alamethecin, sonication, or Brij 58 (polyoxyethylene monocetyl ether). Interestingly, holding the other assay components constant, estradiol-3 glucuronide formation was up to 2.5-fold greater using microsomes prepared in phosphate buffer compared with those prepared in sucrose. Incubations activated by alamethecin consistently exhibited the highest rates of estradiol glucuronidation versus the other activators. Furthermore, estradiol-3 glucuronidation exhibited autoactivation kinetics in all of the conditions investigated (n = 1.2-1.7). Nontraditional kinetics were also observed when other UGT1A1 substrates such as ethinylestradiol, buprenorphine, and anthraflavic acid were studied with both human liver microsomes and recombinant UGT1A1. Naphthol, propofol, morphine, and androstanediol were used as probe UGT substrates selective for UGT1A6, UGT1A9, UGT2B7, and UGT2B15, respectively. Of these substrates, only androstanediol exhibited nontraditional kinetics using human liver microsomes. In conclusion, the Hill and/or Michaelis-Menten equations should be used to fit kinetic data to obtain an accurate assessment of in vitro glucuronidation. PMID- 12756210 TI - Interaction of buprenorphine and its metabolite norbuprenorphine with cytochromes p450 in vitro. AB - Buprenorphine is a thebaine derivative used in the treatment of heroin and other opiate addictions. In this study, the selective probe reactions for each of the major hepatic cytochromes P450 (P450s) were used to evaluate the effect of buprenorphine and its main metabolite norbuprenorphine on the activity of these P450s. The index reactions used were CYP1A2 (phenacetin O-deethylation), CYP2A6 (coumarin 7-hydroxylation), CYP2C9 (diclofenac 4'-hydroxylation), CYP2C19 (omeprazole 5-hydrxoylation), CYP2D6 (dextromethorphan O-demethylation), CYP2B6 (7-ethoxy-4-trifluoromethyl-coumarin 7-deethylation), CYP2E1 (chlorzoxazone 6 hydroxylation), and CYP3A4 (omeprazole sulfoxidation). Buprenorphine exhibited potent, competitive inhibition of CYP2D6 (Ki 10 +/- 2 microM and 1.8 +/- 0.2 microM) and CYP3A4 (Ki 40 +/- 1.6 microM and 19 +/- 1.2 microM) in microsomes from human liver and cDNA-expressing lymphoblasts, respectively. Compared with buprenorphine, norbuprenorphine demonstrated a lower inhibitory potency with CYP2D6 (22.4% inhibition at 20 microM norbuprenorphine) and CYP3A4 (13.6% inhibition at 20 microM) in microsomes from human cDNA-expressing lymphoblast cells. Furthermore, buprenorphine was shown to be a substrate of CYP2D6 (Km = 600 microM; Vmax = 0.40 nmol/min/mg protein) and CYP3A4 (Km = 36 microM; Vmax = 0.19 nmol/min/mg protein). The present in vitro study suggests that buprenorphine and its major metabolite norbuprenorphine are inhibitors of CYP2D6 and CYP3A4; however, at therapeutic concentrations they are not predicted to cause potentially clinically important drug interactions with other drugs metabolized by major hepatic P450s. PMID- 12756211 TI - Induction of hepatic phase II drug-metabolizing enzymes by 1,7-phenanthroline in rats is accompanied by induction of MRP3. AB - The purpose of the present study was to evaluate the effect of 1,7-phenanthroline (PH), which has been proposed to be a selective phase II enzyme inducer, on the gene expression of xenobiotic transporters, as well as hepatic and renal drug metabolizing enzymes. After oral administration of PH for 3 days to male Sprague Dawley rats, mRNA levels in liver (75 and 150 mg/kg doses) and kidney (75 mg/kg dose only) were determined using real-time quantitative polymerase chain reaction. At 150 mg/kg/day, PH treatment resulted in significant increases in hepatic mRNA levels of Mrp3 (36-fold), UGT1A6 (20-fold), UGT2B1 (4-fold), and quinone reductase (QR, 5-fold), compared with the vehicle-treated group. Similar increases in Mrp3 (99-fold), UGT1A6 (17-fold), UGT2B1 (3-fold), and QR (11-fold) mRNA levels were observed in the liver after PH treatment of rats at 75 mg/kg/day. In contrast, the expression levels of CYP2C11 and Oatp2 were decreased by approximately 80 and 50%, respectively. In addition, PH (75 mg/kg/day) elicited statistically significant changes in renal gene expression of CYP3A1, UGT1A6, QR, and Mrp3, but the magnitude of renal Mrp3 induction was less than 2 fold over control. Although PH is known to modulate hepatic glucuronidation in vivo, these data indicated that PH induced mRNA levels of the efflux transporter, Mrp3, which may also affect the disposition of xenobiotics. PMID- 12756212 TI - Effects of acute renal failure on the pharmacokinetics of chlorzoxazone in rats. AB - The purpose of this study is to report the changes of CYP2E1, CYP1A2, CYP2B1/2, CYP2C11, CYP3A23, and CYP3A2 expression and pharmacokinetics and tissue distribution of chlorzoxazone (CZX) and 6-hydroxychlorzoxazone (OH-CZX) in rats with acute renal failure induced by uranyl nitrate (U-ARF), and the role of CYP3A23 and CYP3A2 in the formation of OH-CZX in rats with U-ARF. In rats with U ARF, CYP2C11 decreased to 20% of control, whereas CYP2E1 and CYP3A23 increased 2.3 and 4 times, respectively, compared with control. But expression of CYP1A2 and CYP2B1/2 was not changed by U-ARF. After i.v. administration of CZX at a dose of 20 mg/kg to rats with U-ARF, the areas under the plasma concentration-time curve from time 0 to time infinity (AUCs) of CZX and OH-CZX were significantly smaller and greater, respectively, than those in control rats. In rats with U ARF, CZX was below the detection limit at 120 min in all rat tissues studied, whereas it was detected in all tissues of control rats at both 30 and 120 min. However, in control rats, OH-CZX was below the detection limit at both 30 and 120 min in all rat tissues except kidney, whereas it was detected in all tissues of rats with U-ARF at both 30 and 120 min. Based on results from supporting experiments with DDT and 2,2-bis(4-chlorophenyl)1,1-dichloroethylene treatment of rats, the contribution of CYP3A23 and CYP3A2 to the enhanced formation of OH-CZX in rats with U-ARF is likely to be negligible. PMID- 12756213 TI - Brain penetration of aprepitant, a substance P receptor antagonist, in ferrets. AB - The pharmacokinetics, metabolism, and brain penetration of the neurokinin 1 (NK1) receptor antagonist (substance P receptor antagonist), aprepitant (MK-0869), were examined in ferrets. This species exhibits human-type NK1receptor pharmacology and is of proven value in the identification of clinically useful drugs for the treatment of chemotherapy-induced nausea and vomiting in humans. After a single p.o. dose of aprepitant at 1 or 2 mg/kg, plasma levels of the compound were between approximately 200 and 270 ng/ml, 24 h after dosing. In the brain cortex, concentrations of aprepitant reached between approximately 80 and 150 ng/g of tissue 24 h after dosing. The predominant radioactive component present in the plasma and the brain of ferrets at 24 or 48 h after a single oral dose of [14C]aprepitant at 3 mg/kg was the parent compound itself. The slow plasma clearance of aprepitant ( approximately 1.5 ml/min/kg) and its abundance in ferret brain were in accord with its efficacy in blocking the retching and vomiting at 24 and 48 h postdose when ferrets were challenged with the emetic anticancer drug, cisplatin. When aprepitant and some of its metabolites were assessed for their in vitro binding affinity to the human NK1receptor, aprepitant demonstrated the highest affinity. Collectively, these data suggested that aprepitant, rather than its metabolites, was responsible, primarily, for the antiemetic activity of this compound in the male ferret. PMID- 12756214 TI - Identification of the metabolites of 9-nitro-20(S)-camptothecin in rats. AB - 9-Nitro-20(S)-camptothecin is a novel anticancer drug. In this study, metabolites of 9-nitro-20(S)-camptothecin in rats were identified. Rats were dosed with the drug, and the metabolites in the bile were isolated and collected by high performance liquid chromatography using a gradient elution. By LC/MSn(n =1-3), the biliary metabolites in addition to the unchanged drug were identified as 9 amino-20(S)-camptothecin (M3), 9-acetamido-20(S)-camptothecin (M4), and the glucuronide of 9-hydroxy-20(S)-camptothecin (M1) with the aid of the reference substances and the enzymatic hydrolysis. The accurate mass data for metabolites M2 and M5 were determined by a quadrupole time-of-flight mass spectrometer. Metabolite M5 was suggested to be the glutathione conjugate of 10-hydroxy-20(S) campto-thecin, and M2 may arise from the loss of glutamic acid from M5. Metabolites M1 and M3 were also found in the urine, and M4 in the feces. Fecal metabolite M7 was confirmed to be 9-hydroxy-20(S)-camptothecin (the aglycon of M1) by being compared with the reference standard. The mercapturic acid conjugate of 10-hydroxy-20(S)-camptothecin (M6) was detected in the urine and feces by LC/MSn PMID- 12756215 TI - Hepatobiliary excretion of acetaminophen glutathione conjugate and its derivatives in transport-deficient (TR-) hyperbilirubinemic rats. AB - The involvement of the canalicular multidrug resistance protein 2 (Mrp2) in the hepatobiliary excretion of acetaminophen (APAP)-glutathione (GSH) conjugate and its derivatives was investigated using transport-deficient (TR- rats. Although no differences in the biliary concentration of APAP itself were detected between normal Wistar and TR- rats, significant differences in the biliary disposition of several conjugated metabolites of APAP were detected. APAP-GSH was virtually absent in bile from TR- rats. Also, biliary concentrations of APAP-mercapturate (NAC; N-acetylated l-cysteine) and APAP-GLU were significantly reduced in TR- rats. No differences in the biliary concentration of APAP cysteinylglycine/cysteine (CG/CYS) were detected between normal and mutant rats. The cumulative amounts of APAP-CG/CYS and APAP-NAC excreted in urine of mutant rats were decreased, whereas APAP-GLU was markedly increased. Analysis of liver samples revealed that APAP-GSH and APAP-NAC accumulate in mutant rat livers. Our results support the direct involvement of Mrp2 in the hepatobiliary excretion of several conjugated metabolites of APAP, including APAP-GSH and APAP-NAC, and provide relevant information on processes that may be involved with both their hepatic basolateral transport and renal elimination. PMID- 12756217 TI - Terminology for describing the elastic behavior of arteries. PMID- 12756216 TI - Absorption/metabolism of sulforaphane and quercetin, and regulation of phase II enzymes, in human jejunum in vivo. AB - For the first time the human intestinal effective permeability, estimated from the luminal disappearance and intestinal metabolism of phytochemicals, sulforaphane and quercetin-3,4'-glucoside, as well as the simultaneous changes in gene expression in vivo in enterocytes, has been studied in the human jejunum in vivo (Loc-I-Gut). Both compounds as components of an onion and broccoli extract could readily permeate the enterocytes in the perfused jejunal segment. At the physiologically relevant, dietary concentration tested, the average effective jejunal permeability (Peff) and percentage absorbed (+/- S.D.) were 18.7 +/- 12.6 x 10-4 cm/s and 74 +/- 29% for sulforaphane and 8.9 +/- 7.1 x 10-4 cm/s and 60 +/ 31% for quercetin-3,4'-diglucoside, respectively. Furthermore, a proportion of each compound was conjugated and excreted back into the lumen as sulforaphane glutathione and quercetin-3'-glucuronide. The capacity of the isolated segment to deconjugate quercetin from quercetin-3,4'-diglucoside during the perfusion was much higher than the beta-glucosidase activity of the preperfusion jejunal contents, indicating that the majority (79-100%) of the beta-glucosidase capacity derives from the enterocytes in situ. Simultaneously, we determined short-term changes in gene expression in exfoliated enterocytes, which showed 2.0 +/- 0.4 fold induction of glutathione transferase A1 (GSTA1) mRNA (p < 0.002) and 2.4 +/- 1.2-fold induction of UDP-glucuronosyl transferase 1A1 (UGT1A1) mRNA (p < 0.02). The changes in gene expression were also seen in differentiated Caco-2 cells, where sulforaphane was responsible for induction of GSTA1 and quercetin for induction of UGT1A1. These results show that food components have the potential to modify drug metabolism in the human enterocyte in vivo very rapidly. PMID- 12756219 TI - Effects of the Dietary Approaches to Stop Hypertension (DASH) diet on the pressure-natriuresis relationship. AB - Blood pressure-lowering mechanisms of the Dietary Approaches to Stop Hypertension (DASH) diet, rich in fruits, vegetables, and low-fat dairy foods, were analyzed based on the pressure-natriuresis relationship. Participants (n=375) were randomly assigned to control or DASH diet groups by using a parallel-group design. They then ate their assigned diet for 3 consecutive 30-day intervention feeding periods, during which sodium intake varied among 3 levels by a randomly assigned sequence. Urinary sodium excretion rate and mean arterial pressure were measured at the end of each sodium intake level. Mean arterial pressure and urinary sodium excretion were plotted on x and y axes, respectively, for participants eating control and DASH diets and were modeled as linear relationships for simplicity to allow the estimation of the extrapolated x intercept and slope of the relationships. The DASH diet steepened the slope of the relationship (29.5+/-3.4 vs 64.9+/-13.1 [mmol/d]/mm Hg, P=0.0002) without significantly shifting the x-intercept (94.1+/-0.5 vs 93.2+/-0.6 mm Hg, NS) of the relationship. These data suggest a natriuretic action of the DASH diet. PMID- 12756218 TI - Endothelin-1 and vascular tone in subjects with atherogenic risk factors. AB - Endothelin-1 (ET-1) is a potent vasoconstrictor that increases vascular tone in the resistance vessels of subjects with hypertension. It is unclear whether endogenous ET-1 affects resistance-vessel function equally in patients with other cardiovascular risk factors. Vasoconstriction to ET-1 is mediated principally via the endothelin-A (ETA) receptor on vascular smooth muscle cells. Accordingly, we used an ETA-specific antagonist, BQ-123, to test the hypothesis that endogenous ET-1 increases vascular resistance selectively in subjects with hypertension compared with other risk factors. BQ-123 was infused at 100 nmol/min for 80 minutes into the brachial artery of 10 subjects with hypertension (mean+/-SEM arterial pressure, 106+/-5 mm Hg), 12 subjects with hypercholesterolemia (mean+/ SEM total cholesterol, 7.1+/-0.2 mmol/L), 10 active smokers (mean+/-SEM, 42+/-11 pack-years), and 11 healthy, age-matched individuals. Forearm blood flow (FBF) was measured by venous occlusion plethysmography. BQ-123 dilated resistance arterioles in hypertensive subjects, with FBF's increasing by 46+/-7% from baseline (P<0.001). BQ-123 increased FBF to a lesser extent in hypercholesterolemic (24+/-5%, P<0.001) and healthy (20+/-8%, P=0.007) individuals but did not affect FBF significantly in smokers (10+/-8%, P=0.185). The vasodilator response in hypertensive subjects, but not in hypercholesterolemic patients or smokers, was significantly greater than that in healthy individuals (P=0.012). Endogenous ET-1, acting via the ETA receptor, increases resistance-vessel tone in subjects with hypertension more than in subjects with hypercholesterolemia or in smokers. These results indicate that ET 1 contributes more to the pathophysiology of hypertension than of other risk factors in subjects without overt atherosclerosis. PMID- 12756220 TI - Effects of pressure overload on extracellular matrix expression in the heart of the atrial natriuretic peptide-null mouse. AB - This study tested the hypothesis that atrial natriuretic peptide has direct antihypertrophic actions on the heart by modulating expression of genes involved in cardiac hypertrophy and extracellular matrix production. Hearts of male, atrial natriuretic peptide-null and control wild-type mice that had been subjected to pressure overload after transverse aortic constriction and control unoperated hearts were weighed and subjected to microarray, Northern blot, and immunohistochemical analyses. Microarray and Northern blot analyses were used to identify genes that are regulated differentially in response to stress in the presence and absence of atrial natriuretic peptide. Immunohistochemical analysis was used to identify and localize expression of the protein products of these genes. Atrial natriuretic peptide-null mice demonstrated cardiac hypertrophy at baseline and an exaggerated hypertrophic response to transverse aortic constriction associated with increased expression of the extracellular matrix molecules periostin, osteopontin, collagen I and III, and thrombospondin, as well as the extracellular matrix regulatory proteins, matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-3, and the novel growth factor pleiotrophin compared with wild-type controls. These results support the hypothesis that atrial natriuretic peptide protects against pressure overload-induced cardiac hypertrophy and remodeling by negative modulation of genes involved in extracellular matrix deposition. PMID- 12756221 TI - Mild renal dysfunction and subclinical cardiovascular damage in primary hypertension. AB - The presence of mild renal dysfunction is associated with high cardiovascular morbidity and mortality rates in patients with primary hypertension. The pathophysiological mechanisms underlying this association are currently unknown. We investigated the relation between mild renal dysfunction and subclinical cardiovascular organ damage in 358 never previously treated patients with primary hypertension. Mild renal dysfunction was defined as a creatinine clearance <60 mL/min and/or the presence of microalbuminuria. Left ventricular mass index and carotid intima-media thickness were assessed by ultrasound scan. The prevalence of mild renal dysfunction, left ventricular hypertrophy, and carotid plaque was 18%, 48%, and 28%, respectively. Mild renal dysfunction was related to the presence of several risk factors, such as older age, higher blood pressure levels and lipid status, and smoking habits. Patients with the highest left ventricular mass and carotid intima-media thickness (upper quartiles) showed a higher prevalence of mild renal dysfunction (P<0.0001). After adjusting for duration of hypertension, mean blood pressure, smoking habits, and age, we found that the risk of left ventricular hypertrophy and/or carotid atherosclerosis increased by 43% with each SD reduction in creatinine clearance, and by 89% with each SD increase in albuminuria. Mild renal dysfunction is associated with preclinical end-organ damage in patients with primary hypertension. These data may help explain the high cardiovascular mortality rates reported in patients with low glomerular filtration rate or with increased albuminuria. The evaluation of creatinine clearance and urinary albumin excretion could be useful for identifying subjects at higher cardiovascular risk. PMID- 12756222 TI - Seventh report of the Joint National Committee on the Prevention, Detection, Evaluation, and Treatment of High Blood Pressure (JNC 7): resetting the hypertension sails. PMID- 12756223 TI - Blood pressure indices and cardiovascular disease in the Asia Pacific region: a pooled analysis. AB - This article aims to compare the importance of systolic blood pressure (SBP), diastolic blood pressure (DBP), mean arterial pressure (MAP), and pulse pressure (PP) as risk factors for stroke and ischemic heart disease and to assess whether the patterns are consistent by age and gender. Cox proportional-hazards regression, adjusted for cholesterol and smoking, was used to assess the associations of the 4 BP indices with stroke and ischemic heart disease by age and gender. The relative importance of individual indices was assessed with a hazard ratios for a 1-SD change in BP and by likelihood-ratio chi2 tests. The influence of >1 BP index in the Cox model was also estimated. The analyses demonstrated similar associations of SBP, DBP, and MAP with both fatal stroke and ischemic heart diseases, which were stronger than those of PP. Both SBP and MAP tended to be more important in the regression model than DBP or PP. In Cox models including DBP, addition of SBP improved the goodness of fit at all ages and for both genders. However, in Cox models including SBP, addition of DBP typically resulted in little incremental benefit over and above that of SBP alone. These data suggest that if time or resources are highly constrained, such as in much needed epidemiologic surveys in developing countries, very little is lost from only measuring SBP. PMID- 12756224 TI - Stem cells in the skin: waste not, Wnt not. PMID- 12756225 TI - Mice lacking a transcriptional corepressor Tob are predisposed to cancer. AB - tob is a member of antiproliferative family genes. Mice lacking tob are prone to spontaneous formation of tumors. The occurrence rate of diethylnitrosamine induced liver tumors is higher in tob(-/-) mice than in wild-type mice. tob(-/ )p53(-/-) mice show accelerated tumor formation in comparison with single null mice. Expression of cyclin D1 mRNA is increased in the absence of Tob and is reduced by Tob. Tob acts as a transcriptional corepressor and suppresses the cyclin D1 promoter activity through an interaction with histone deacetylase. Levels of tob mRNA are often decreased in human cancers, implicating tob in cancer development. PMID- 12756226 TI - Transient activation of beta -catenin signaling in cutaneous keratinocytes is sufficient to trigger the active growth phase of the hair cycle in mice. AB - Wnts have key roles in many developmental processes, including hair follicle growth and differentiation. Stabilization of beta-catenin is essential in the canonical Wnt signaling pathway. We developed transgenic mice expressing a regulated form of beta-catenin in the skin. Chronic activation of beta-catenin in resting (telogen) hair follicles resulted in changes consistent with induction of an exaggerated, aberrant growth phase (anagen). Transient activation of beta catenin produced a normal anagen. Our data lend strong support to the notion that a Wnt/beta-catenin signal operating on hair follicle precursor cells serves as a crucial proximal signal for the telogen-anagen transition. PMID- 12756227 TI - In vitro propagation and transcriptional profiling of human mammary stem/progenitor cells. AB - Although the existence of mammary stem cells has been suggested by serial transplantation studies in mice, their identification has been hindered by the lack of specific surface markers, and by the absence of suitable in vitro assays for testing stem cell properties: self-renewal and ability to generate differentiated progeny. We have developed an in vitro cultivation system that allows for propagation of human mammary epithelial cells (HMECs) in an undifferentiated state, based on their ability to proliferate in suspension, as nonadherent mammospheres. We demonstrate that nonadherent mammospheres are enriched in early progenitor/stem cells and able to differentiate along all three mammary epithelial lineages and to clonally generate complex functional structures in reconstituted 3D culture systems. Gene expression analysis of cells isolated from nonadherent mammospheres revealed overlapping genetic programs with other stem and progenitor cells and identified new markers that may be useful in the identification of mammary stem cells. The isolation and characterization of these stem cells should help elucidate the molecular pathways that govern normal mammary development and carcinogenesis. PMID- 12756228 TI - JNK initiates a cytokine cascade that causes Pax2 expression and closure of the optic fissure. AB - The c-Jun NH(2)-terminal kinase (JNK) group of mitogen-activated protein kinases is stimulated in response to a wide array of cellular stresses and proinflammatory cytokines. Mice lacking individual members of the Jnk family (Jnk1, Jnk2, and Jnk3) are viable and survive without overt structural abnormalities. Here we show that mice with a compound deficiency in Jnk expression can survive to birth, but fail to close the optic fissure (retinal coloboma). We demonstrate that JNK initiates a cytokine cascade of bone morphogenetic protein-4 (BMP4) and sonic hedgehog (Shh) that induces the expression of the paired-like homeobox transcription factor Pax2 and closure of the optic fissure. Interestingly, the role of JNK to regulate BMP4 expression during optic fissure closure is conserved in Drosophila during dorsal closure, a related morphogenetic process that requires JNK-regulated expression of the BMP4 ortholog Decapentaplegic (Dpp). PMID- 12756229 TI - RNA polymerase mutations that impair conversion to a termination-resistant complex by Q antiterminator proteins. AB - Bacteriophage lambda Q-protein stably binds and modifies RNA polymerase (RNAP) to a termination-resistant form. We describe amino acid substitutions in RNAP that disrupt Q-mediated antitermination in vivo and in vitro. The positions of these substitutions in the modeled RNAP/DNA/RNA ternary elongation complex, and their biochemical properties, suggest that they do not define a binding site for Q in RNAP, but instead act by impairing interactions among core RNAP subunits and nucleic acids that are essential for Q modification. A specific conjecture is that Q modification stabilizes interactions of RNAP with the DNA/RNA hybrid and optimizes alignment of the nucleic acids in the catalytic site. Such changes would inhibit the activity of the RNA hairpin of an intrinsic terminator to disrupt the 5'-terminal bases of the hybrid and remove the RNA 3' terminus from the active site. PMID- 12756230 TI - An intersubunit contact stimulating transcription initiation by E coli RNA polymerase: interaction of the alpha C-terminal domain and sigma region 4. AB - The C-terminal domain of the Escherichia coli RNA polymerase (RNAP) alpha subunit (alphaCTD) stimulates transcription initiation by interacting with upstream (UP) element DNA and a variety of transcription activators. Here we identify specific substitutions in region 4.2 of sigma 70 (sigma(70)) and in alphaCTD that decrease transcription initiation from promoters containing some, but not all, UP elements. This decrease in transcription derives from a decrease in the initial equilibrium constant for RNAP binding (K(B)). The open complexes formed by the mutant and wild-type RNAPs differ in DNAse I sensitivity at the junction of the alphaCTD and sigma DNA binding sites, correlating with the differences in transcription. A model of the DNA-alphaCTD-sigma region 4.2 ternary complex, constructed from the previously determined X-ray structures of the Thermus aquaticus sigma region 4.2-DNA complex and the E. coli alphaCTD-DNA complex, indicates that the residues identified by mutation in sigma region 4.2 and in alphaCTD are in very close proximity. Our results strongly suggest that alphaCTD, when bound to an UP element proximal subsite, contacts the RNAP sigma(70) subunit, increasing transcription. Previous data from the literature suggest that this same sigma-alphaCTD interaction also plays a role in transcription factor mediated activation. PMID- 12756231 TI - Heterochromatin protein 1 (HP1) is associated with induced gene expression in Drosophila euchromatin. AB - Heterochromatin protein 1 (HP1) is a conserved nonhistone chromosomal protein, which is involved in heterochromatin formation and gene silencing in many organisms. In addition, it has been shown that HP1 is also involved in telomere capping in Drosophila. Here, we show a novel striking feature of this protein demonstrating its involvement in the activation of several euchromatic genes in Drosophila. By immunostaining experiments using an HP1 antibody, we found that HP1 is associated with developmental and heat shock-induced puffs on polytene chromosomes. Because the puffs are the cytological phenotype of intense gene activity, we did a detailed analysis of the heat shock-induced expression of the HSP70 encoding gene in larvae with different doses of HP1 and found that HP1 is positively involved in Hsp70 gene activity. These data significantly broaden the current views of the roles of HP1 in vivo by demonstrating that this protein has multifunctional roles. PMID- 12756232 TI - The Caenorhabditis elegans vab-10 spectraplakin isoforms protect the epidermis against internal and external forces. AB - Morphogenesis of the Caenorhabditis elegans embryo is driven by actin microfilaments in the epidermis and by sarcomeres in body wall muscles. Both tissues are mechanically coupled, most likely through specialized attachment structures called fibrous organelles (FOs) that connect muscles to the cuticle across the epidermis. Here, we report the identification of new mutations in a gene known as vab-10, which lead to severe morphogenesis defects, and show that vab-10 corresponds to the C. elegans spectraplakin locus. Our analysis of vab-10 reveals novel insights into the role of this plakin subfamily. vab-10 generates isoforms related either to plectin (termed VAB-10A) or to microtubule actin cross linking factor plakins (termed VAB-10B). Using specific antibodies and mutations, we show that VAB-10A and VAB-10B have distinct distributions and functions in the epidermis. Loss of VAB-10A impairs the integrity of FOs, leading to epidermal detachment from the cuticle and muscles, hence demonstrating that FOs are functionally and molecularly related to hemidesmosomes. We suggest that this isoform protects against forces external to the epidermis. In contrast, lack of VAB-10B leads to increased epidermal thickness during embryonic morphogenesis when epidermal cells change shape. We suggest that this isoform protects cells against tension that builds up within the epidermis. PMID- 12756233 TI - Interplay of signal recognition particle and trigger factor at L23 near the nascent chain exit site on the Escherichia coli ribosome. AB - As newly synthesized polypeptides emerge from the ribosome, they interact with chaperones and targeting factors that assist in folding and targeting to the proper location in the cell. In Escherichia coli, the chaperone trigger factor (TF) binds to nascent polypeptides early in biosynthesis facilitated by its affinity for the ribosomal proteins L23 and L29 that are situated around the nascent chain exit site on the ribosome. The targeting factor signal recognition particle (SRP) interacts specifically with the signal anchor (SA) sequence in nascent inner membrane proteins (IMPs). Here, we have used photocross-linking to map interactions of the SA sequence in a short, in vitro-synthesized, nascent IMP. Both TF and SRP were found to interact with the SA with partially overlapping binding specificity. In addition, extensive contacts with L23 and L29 were detected. Both purified TF and SRP could be cross-linked to L23 on nontranslating ribosomes with a competitive advantage for SRP. The results suggest a role for L23 in the targeting of IMPs as an attachment site for TF and SRP that is close to the emerging nascent chain. PMID- 12756234 TI - Photocross-linking of nascent chains to the STT3 subunit of the oligosaccharyltransferase complex. AB - In eukaryotic cells, polypeptides are N glycosylated after passing through the membrane of the ER into the ER lumen. This modification is effected cotranslationally by the multimeric oligosaccharyltransferase (OST) enzyme. Here, we report the first cross-linking of an OST subunit to a nascent chain that is undergoing translocation through, or integration into, the ER membrane. A photoreactive probe was incorporated into a nascent chain using a modified Lys tRNA and was positioned in a cryptic glycosylation site (-Q-K-T- instead of -N-K T-) in the nascent chain. When translocation intermediates with nascent chains of increasing length were irradiated, nascent chain photocross-linking to translocon components, Sec61alpha and TRAM, was replaced by efficient photocross-linking solely to a protein identified by immunoprecipitation as the STT3 subunit of the OST. No cross-linking was observed in the absence of a cryptic sequence or in the presence of a competitive peptide substrate of the OST. As no significant nascent chain photocross-linking to other OST subunits was detected in these fully assembled translocation and integration intermediates, our results strongly indicate that the nascent chain portion of the OST active site is located in STT3. PMID- 12756235 TI - Colocalization of synapsin and actin during synaptic vesicle recycling. AB - It has been hypothesized that in the mature nerve terminal, interactions between synapsin and actin regulate the clustering of synaptic vesicles and the availability of vesicles for release during synaptic activity. Here, we have used immunogold electron microscopy to examine the subcellular localization of actin and synapsin in the giant synapse in lamprey at different states of synaptic activity. In agreement with earlier observations, in synapses at rest, synapsin immunoreactivity was preferentially localized to a portion of the vesicle cluster distal to the active zone. During synaptic activity, however, synapsin was detected in the pool of vesicles proximal to the active zone. In addition, actin and synapsin were found colocalized in a dynamic filamentous cytomatrix at the sites of synaptic vesicle recycling, endocytic zones. Synapsin immunolabeling was not associated with clathrin-coated intermediates but was found on vesicles that appeared to be recycling back to the cluster. Disruption of synapsin function by microinjection of antisynapsin antibodies resulted in a prominent reduction of the cytomatrix at endocytic zones of active synapses. Our data suggest that in addition to its known function in clustering of vesicles in the reserve pool, synapsin migrates from the synaptic vesicle cluster and participates in the organization of the actin-rich cytomatrix in the endocytic zone during synaptic activity. PMID- 12756236 TI - The Sec1p/Munc18 (SM) protein, Vps45p, cycles on and off membranes during vesicle transport. AB - Protein phosphatase 1 (PP1, Glc7p) functions in the final stage of SNARE-mediated vesicle transport between docking and fusion. During this process, trans-SNARE complexes, formed between molecules in opposing membranes, convert to cis complexes, with all participants in the same lipid bilayer. Here, we show that glc7 mutant cells accumulate SNARE complexes. These complexes are clearly different from those found in either wild-type or sec18-1 cells as the Sec1p/Munc18 (SM) protein Vps45p does not bind to them. Given that PP1 controls fusion, the SNARE complexes that accumulate in glc7 mutants likely represent trans-SNARE complexes. Vps45p dissociates from the membrane in the absence of PP1 activity, but rapidly reassociates after its reactivation. These data reveal that SM proteins cycle on and off membranes in a stage-specific manner during the vesicle transport reaction, and suggest that protein phosphorylation plays a key role in the regulation of this cycle. PMID- 12756237 TI - Macrophage podosomes assemble at the leading lamella by growth and fragmentation. AB - Podosomes are actin- and fimbrin-containing adhesions at the leading edge of macrophages. In cells transfected with beta-actin-ECFP and L-fimbrin-EYFP, quantitative four-dimensional microscopy of podosome assembly shows that new adhesions arise at the cell periphery by one of two mechanisms; de novo podosome assembly, or fission of a precursor podosome into daughter podosomes. The large podosome cluster precursor also appears to be an adhesion structure; it contains actin, fimbrin, integrin, and is in close apposition to the substratum. Microtubule inhibitors paclitaxel and demecolcine inhibit the turnover and polarized formation of podosomes, but not the turnover rate of actin in these structures. Because daughter podosomes and podosome cluster precursors are preferentially located at the leading edge, they may play a critical role in continually generating new sites of cell adhesion. PMID- 12756239 TI - Calcium-dependent protein interactions in MUC5B provide reversible cross-links in salivary mucus. AB - The macromolecular organization within saliva was investigated by tracer diffusion measurements of fluorescent polystyrene microspheres by fluorescence recovery after photobleaching using a confocal microscope (confocal-FRAP). There was a concentration-dependent reduction in microsphere diffusion; this was much greater in the presence of calcium (10 mm) and was reduced by the addition of EGTA (10 mm). These effects on tracer diffusion showed that native saliva contained a macromolecular organization that was sensitive to free calcium concentrations. This was supported by a major increase in the weight average molecular weight of the high molecular weight mucin fraction in saliva (10-62 x 106) and an increase in intrinsic viscosity of saliva (733 to 1203 ml/g) both caused by calcium. Analysis of the change in tracer diffusion in saliva showed a 20-fold increase in the apparent pore size (from 130 nm in 10 mm CaCl2 to 2600 nm in 10 mm EGTA at physiological concentration). The effect was specific for calcium and was unaffected by up to 2 m NaCl. The calcium binding activity was contained in a high buoyant density fraction of saliva excluded from Sepharose CL 2B. Calcium binding to this fraction gave an approximate Kd of 7 x 10-6 m, and the binding was irreversibly destroyed by treatment with 6 m guanidinium chloride and by mild reduction, suggesting it to be to a protein site. This fraction of saliva was shown to contain MUC5B as the single major protein species by positive ion electrospray ionization-tandem mass spectrometry analysis. The results suggested that oligomeric MUC5B in saliva is assembled into much larger linear or branched assemblies through calcium-mediated protein cross-links. PMID- 12756238 TI - A novel pathway for MuSK to induce key genes in neuromuscular synapse formation. AB - At the developing neuromuscular junction the Agrin receptor MuSK is the central organizer of subsynaptic differentiation induced by Agrin from the nerve. The expression of musk itself is also regulated by the nerve, but the mechanisms involved are not known. Here, we analyzed the activation of a musk promoter reporter construct in muscle fibers in vivo and in cultured myotubes, using transfection of multiple combinations of expression vectors for potential signaling components. We show that neuronal Agrin by activating MuSK regulates the expression of musk via two pathways: the Agrin-induced assembly of muscle derived neuregulin (NRG)-1/ErbB, the pathway thought to regulate acetylcholine receptor (AChR) expression at the synapse, and via a direct shunt involving Agrin induced activation of Rac. Both pathways converge onto the same regulatory element in the musk promoter that is also thought to confer synapse-specific expression to AChR subunit genes. In this way, a positive feedback signaling loop is established that maintains musk expression at the synapse when impulse transmission becomes functional. The same pathways are used to regulate synaptic expression of AChR epsilon. We propose that the novel pathway stabilizes the synapse early in development, whereas the NRG/ErbB pathway supports maintenance of the mature synapse. PMID- 12756240 TI - Ssq1, a mitochondrial Hsp70 involved in iron-sulfur (Fe/S) center biogenesis. Similarities to and differences from its bacterial counterpart. AB - The results of in vivo and in organellar experiments indicate that the Hsp70 Ssq1 and the J-protein Jac1 function together to assist in the biogenesis of iron sulfur (Fe/S) centers in the mitochondrial matrix. Here we present biochemical evidence supporting this idea. Isu, the proposed scaffold on which Fe/S centers are assembled, is a substrate for both Jac1 and Ssq1. Jac1 and Isu1 cooperatively stimulate the ATPase activity of Ssq1. In addition, Jac1 facilitates the interaction of Ssq1 with Isu1 in the presence of ATP. These findings are consistent with the role in Fe/S biogenesis previously proposed for the bacterial Hsp70 Hsc66 and J-protein Hsc20 that interact with the bacterial Isu homologue IscU. However, unlike the bacterial Hsp70, we found that Ssq1 has a high affinity for nucleotide, and shares a nucleotide exchange factor, Mge1, with a second mitochondrial Hsp70, Ssc1. Thus, whereas the bacterial and mitochondrial chaperone systems share critical features, they possess significant biochemical differences as well. PMID- 12756241 TI - Reverse reaction of lysophosphatidylinositol acyltransferase. Functional reconstitution of coenzyme A-dependent transacylation system. AB - CoA-dependent transacylation activity in microsomes catalyzes the transfer of fatty acid between phospholipids and lysophospholipids in the presence of CoA without the generation of free fatty acid. We examined the mechanism of the transacylation system using partially purified acyl-CoA:lysophosphatidylinositol (LPI) acyltransferase (LPIAT) from rat liver microsomes to test our hypothesis that both the reverse and forward reactions of acyl-CoA:lysophospholipid acyltransferases are involved in the CoA-dependent transacylation process. The purified LPIAT fraction exhibited ATP-independent acyl-CoA synthetic activity and CoA-dependent LPI generation from PI, suggesting that LPIAT could operate in reverse to form acyl-CoA and LPI. CoA-dependent acylation of LPI by the purified LPIAT fraction required PI as the acyl donor. In addition, the combination of purified LPIAT and recombinant lysophosphatidic acid acyltransferase could reconstitute CoA-dependent transacylation between PI and phosphatidic acid. These results suggest that the CoA-dependent transacylation system consists of the following: 1) acyl-CoA synthesis from phospholipid through the reverse action of acyl-CoA:lysophospholipid acyltransferases; and 2) transfer of fatty acyl moiety from the newly formed acyl-CoA to lysophospholipid through the forward action of acyl-CoA:lysophospholipid acyltransferases. PMID- 12756242 TI - Changed energy state and increased mitochondrial beta-oxidation rate in liver of rats associated with lowered proton electrochemical potential and stimulated uncoupling protein 2 (UCP-2) expression: evidence for peroxisome proliferator activated receptor-alpha independent induction of UCP-2 expression. AB - Lowering of plasma triglyceride levels by hypolipidemic agents is caused by a shift in the liver cellular metabolism, which become poised toward peroxisome proliferator-activated receptor (PPAR) alpha-regulated fatty acid catabolism in mitochondria. After dietary treatment of rats with the hypolipidemic, modified fatty acid, tetradecylthioacetic acid (TTA), the energy state parameters of the liver were altered at the tissue, cell, and mitochondrial levels. Thus, the hepatic phosphate potential, energy charge, and respiratory control coefficients were lowered, whereas rates of oxygen uptake, oxidation of pyridine nucleotide redox pairs, beta-oxidation, and ketogenesis were elevated. Moderate uncoupling of mitochondria from TTA-treated rats was confirmed, as the proton electrochemical potential (Delta(p)) was 15% lower than controls. The change affected the Delta(Psi) component only, leaving the (Delta)pH component unaltered, suggesting that TTA causes induction of electrogenic ion transport rather than electrophoretic fatty acid activity. TTA treatment induced expression of hepatic uncoupling protein 2 (UCP-2) in rats as well as in wild type and PPARalpha-deficient mice, accompanied by a decreased double bond index of the mitochondrial membrane lipids. However, changes of mitochondrial fatty acid composition did not seem to be related to the effects on mitochondrial energy conductance. As TTA activates PPARdelta, we discuss how this subtype might compensate for deficiency of PPARalpha. The overall changes recorded were moderate, making it likely that liver metabolism can maintain its function within the confines of its physiological regulatory framework where challenged by a hypolipemic agent such as TTA, as well as others. PMID- 12756243 TI - Inhibition of calcium uptake via the sarco/endoplasmic reticulum Ca2+-ATPase in a mouse model of Sandhoff disease and prevention by treatment with N butyldeoxynojirimycin. AB - Gangliosides are found at high levels in neuronal tissues where they play a variety of important functions. In the gangliosidoses, gangliosides accumulate because of defective activity of the lysosomal proteins responsible for their degradation, usually resulting in a rapidly progressive neurodegenerative disease. However, the molecular mechanism(s) leading from ganglioside accumulation to neurodegeneration is not known. We now examine the effect of ganglioside GM2 accumulation in a mouse model of Sandhoff disease (one of the GM2 gangliosidoses), the Hexb-/- mouse. Microsomes from Hexb-/- mouse brain showed a significant reduction in the rate of Ca2+-uptake via the sarco/endoplasmic reticulum Ca2+-ATPase (SERCA), which was prevented by feeding Hexb-/- mice with N butyldeoxynojirimycin (NB-DNJ), an inhibitor of glycolipid synthesis that reduces GM2 storage. Changes in SERCA activity were not due to transcriptional regulation but rather because of a decrease in Vmax. Moreover, exogenously added GM2 had a similar effect on SERCA activity. The functional significance of these findings was established by the enhanced sensitivity of neurons cultured from embryonic Hexb-/- mice to cell death induced by thapsigargin, a specific SERCA inhibitor, and by the enhanced sensitivity of Hexb-/- microsomes to calcium-induced calcium release. This study suggests a mechanistic link among GM2 accumulation, reduced SERCA activity, and neuronal cell death, which may be of significance for delineating the neuropathophysiology of Sandhoff disease. PMID- 12756244 TI - Catalytic mechanism revealed by the crystal structure of undecaprenyl pyrophosphate synthase in complex with sulfate, magnesium, and triton. AB - Undecaprenyl pyrophosphate synthase (UPPs) catalyzes chain elongation of farnesyl pyrophosphate (FPP) to undecaprenyl pyrophosphate (UPP) via condensation with eight isopentenyl pyrophosphates (IPP). UPPs from Escherichia coli is a dimer, and each subunit consists of 253 amino acid residues. The chain length of the product is modulated by a hydrophobic active site tunnel. In this paper, the crystal structure of E. coli UPPs was refined to 1.73 A resolution, which showed bound sulfate and magnesium ions as well as Triton X-100 molecules. The amino acid residues 72-82, which encompass an essential catalytic loop not seen in the previous apoenzyme structure (Ko, T.-P., Chen, Y. K., Robinson, H., Tsai, P. C., Gao, Y.-G., Chen, A. P.-C., Wang, A. H.-J., and Liang, P.-H. (2001) J. Biol. Chem. 276, 47474-47482), also became visible in one subunit. The sulfate ions suggest locations of the pyrophosphate groups of FPP and IPP in the active site. The Mg2+ is chelated by His-199 and Glu-213 from different subunits and possibly plays a structural rather than catalytic role. However, the metal ion is near the IPP-binding site, and double mutation of His-199 and Glu-213 to alanines showed a remarkable increase of Km value for IPP. Inside the tunnel, one Triton surrounds the top portion of the tunnel, and the other occupies the bottom part. These two Triton molecules may mimic the hydrocarbon moiety of the UPP product in the active site. Kinetic analysis indicated that a high concentration (>1%) of Triton inhibits the enzyme activity. PMID- 12756245 TI - Identification, purification, and characterization of an eukaryotic-like phosphopantetheine adenylyltransferase (coenzyme A biosynthetic pathway) in the hyperthermophilic archaeon Pyrococcus abyssi. AB - Although coenzymeA (CoA) is essential in numerous metabolic pathways in all living cells, molecular characterization of the CoA biosynthetic pathway in Archaea remains undocumented. Archaeal genomes contain detectable homologues for only three of the five steps of the CoA biosynthetic pathway characterized in Eukarya and Bacteria. In case of phosphopantetheine adenylyltransferase (PPAT) (EC 2.7.7.3), the putative archaeal enzyme exhibits significant sequence similarity only with its eukaryotic homologs, an unusual situation for a protein involved in a central metabolic pathway. We have overexpressed in Escherichia coli, purified, and characterized this putative PPAT from the hyperthermophilic archaeon Pyrococcus abyssi (PAB0944). Matrix-assisted laser desorption ionization time of flight mass spectrometry and high performance liquid chromatography measurements are consistent with the presence of a dephospho-CoA (dPCoA) molecule tightly bound to the polypeptide. The protein indeed catalyzes the synthesis of dPCoA from 4'-phosphopantetheine and ATP, as well as the reverse reaction. The presence of dPCoA stabilizes PAB0944, as it induces a shift from 76 to 82 degrees C of the apparent Tm measured by differential scanning microcalorimetry. Potassium glutamate was found to stabilize the protein at 400 mm. The enzyme behaves as a monomeric protein. Although only distantly related, secondary structure prediction indicates that archaeal and eukaryal PPAT belong to the same nucleotidyltransferase superfamily of bacterial PPAT. The existence of operational proteins highly conserved between Archaea and Eukarya involved in a central metabolic pathway challenge evolutionary scenarios in which eukaryal operational proteins are strictly of bacterial origin. PMID- 12756246 TI - The interaction between HIV-1 Gag and human lysyl-tRNA synthetase during viral assembly. AB - Human lysyl-tRNA synthetase (LysRS) is a tRNA-binding protein that is selectively packaged into HIV-1 along with its cognate tRNALys isoacceptors. Evidence exists that Gag alone is sufficient for the incorporation of LysRS into virions. Herein, using both in vitro and in vivo methods, we begin to map regions in Gag and LysRS that are required for this interaction. In vitro reactions between wild-type and truncated HIV-1 Gag and human LysRS were monitored using GST-tagged molecules and glutathione-agarose chromatography. Gag/LysRS interaction in vivo was detected in 293FT cells cotransfected with plasmids coding for wild-type or mutant HIV-1 Gag and LysRS, either by monitoring Gag.LysRS complexes immunoprecipitated from cell lysate with anti-LysRS or by measuring the ability of LysRS to be packaged into budded Gag viral-like particles. Based on these studies, we conclude that the Gag/LysRS interaction depends upon Gag sequences within the C-terminal domain of capsid (the last 54 amino acids) and amino acids 208-259 of LysRS. The latter domain includes the class II aminoacyl-tRNA synthetase consensus sequence known as motif 1. Both regions have been implicated in homodimerization of capsid and LysRS, respectively. Sequences falling outside these amino acid stretches can be deleted from either molecule without affecting the Gag/LysRS interaction, further supporting the observation that LysRS is incorporated into Gag viral-like particles independent of its ability to bind tRNALys. PMID- 12756247 TI - Regulatory interactions between the checkpoint kinase Chk1 and the proteins of the DNA-dependent protein kinase complex. AB - Checkpoints are biochemical pathways that provide cells a mechanism to detect DNA damage and respond by arresting the cell cycle to allow DNA repair. The conserved checkpoint kinase, Chk1, regulates mitotic progression in response to DNA damage by blocking the activation of Cdk1/cyclin B. In this study, we investigate the regulatory interaction between Chk1 and members of the Atm family of kinases and the functional role of the C-terminal non-catalytic domains of Chk1. Chk1 stimulates the kinase activity of DNA-PK (protein kinase) complexes, which leads to increased phosphorylation of p53 on Ser-15 and Ser-37. In addition, Chk1 stimulates DNA-PK-dependent end-joining reactions in vitro. We also show that Chk1 protein complexes bind to single-stranded DNA and DNA ends. These results indicate a connection between components that regulate the checkpoint pathways and DNA-PK complex proteins, which have a role in the repair of double strand breaks. PMID- 12756248 TI - BLOC-3, a protein complex containing the Hermansky-Pudlak syndrome gene products HPS1 and HPS4. AB - The Hermansky-Pudlak syndrome (HPS) is a genetic disorder characterized by defective lysosome-related organelles. HPS results from mutations in either one of six human genes named HPS1 to HPS6, most of which encode proteins of unknown function. Here we report that the human HPS1 and HPS4 proteins are part of a complex named BLOC-3 (for biogenesis of lysosome-related organelles complex 3). Co-immunoprecipitation experiments demonstrated that epitope-tagged and endogenous HPS1 and HPS4 proteins assemble with each other in vivo. The HPS1.HPS4 complex is predominantly cytosolic, with a small amount being peripherally associated with membranes. Size exclusion chromatography and sedimentation velocity analyses of the cytosolic fraction indicate that HPS1 and HPS4 form a moderately asymmetric protein complex with a molecular mass of approximately 175 kDa. HPS4-deficient fibroblasts from light ear mice display normal distribution and trafficking of the lysosomal membrane protein, Lamp-2, in contrast to fibroblasts from AP-3-deficient pearl mice (HPS2), which exhibit increased trafficking of this lysosomal protein via the plasma membrane. Similarly, light ear fibroblasts display an apparently normal accumulation of Zn2+ in intracellular vesicles, unlike pearl fibroblasts, which exhibit a decreased intracellular Zn2+ storage. Taken together, these observations demonstrate that the HPS1 and HPS4 proteins are components of a cytosolic complex that is involved in the biogenesis of lysosomal-related organelles by a mechanism distinct from that operated by AP-3 complex. PMID- 12756249 TI - Regulation of blastocyst migration, apposition, and initial adhesion by a chemokine, interferon gamma-inducible protein 10 kDa (IP-10), during early gestation. AB - For a pregnancy to be established, initial apposition and adhesion of the blastocyst to maternal endometrium must occur in a coordinated manner; however, a key factor(s) that mediates the trophoblast cell migration and attachment to the apical surface of the endometrium has not been identified. In this study, we examined the effect of an endometrial chemokine, interferon-gamma-inducible protein 10 kDa (IP-10), on conceptus migration to the endometrial epithelium. We first studied endometrial IP-10 mRNA expression, which was localized in the subepithelial stromal region, and detected the protein in the uterine flushing media during early pregnancy. Expression of IP-10 mRNA by the endometrium of cyclic animals was stimulated by the addition of a conceptus factor interferon tau (IFN-tau). Immunofluorescent analysis revealed that IP-10 receptor, CXCR3, was localized in the trophoblast cells, to which biotinylated-recombinant caprine IP-10 (rcIP-10) bound. Chemotaxis assay indicated that rcIP-10 stimulated the migration of trophoblast cells, and the effects of rcIP-10 were neutralized by the pretreatment with an anti-IP-10 antibody. Adhesive activity of trophoblast cells to fibronectin was promoted by rcIP-10, and the effect was inhibited by the use of anti-IP-10 antibody. Further adhesion experiments demonstrated that binding of trophoblast cells to fibronectin was completely inhibited by a peptide of the Arg-Gly-Asp (RGD) sequence, which binds to integrins alpha5beta1, alphaVbeta1, alphaVbeta3, and alphaVbeta5, whereas non-binding peptide containing Arg-Gly-Glu (RGE) had minimal effects. More importantly, rcIP-10 promoted the adhesion of trophoblast cells to primary cells isolated from endometrial epithelium. Furthermore, rcIP-10 stimulated the expression of integrin alpha5, alphaV, and beta3 subunit mRNA in trophoblast cells. These findings suggest that endometrial IP-10 regulates the establishment of apical interactions between trophoblast and epithelial cells during early gestation. PMID- 12756250 TI - Aft1p and Aft2p mediate iron-responsive gene expression in yeast through related promoter elements. AB - The transcription factors Aft1p and Aft2p from Saccharomyces cerevisiae regulate the expression of genes that are involved in iron homeostasis. In vitro studies have shown that both transcription factors bind to an iron-responsive element (FeRE) that is present in the upstream region of genes in the iron regulon. We have used DNA microarrays to distinguish the genes that are activated by Aft1p and Aft2p and to establish for the first time that each factor gives rise to a unique transcriptional profile due to the differential expression of individual iron-regulated genes. We also show that both Aft1p and Aft2p mediate the in vivo expression of FET3 and FIT3 through a consensus FeRE. In addition, both proteins regulate MRS4 via a variant FeRE with Aft2p being the stronger activator from this particular element. Like other paralogous pairs of transcription factors within S. cerevisiae, Aft1p and Aft2p are able to interact with the same promoter elements while maintaining specificity of gene activation. PMID- 12756251 TI - Agonist-independent desensitization and internalization of the human platelet activating factor receptor by coumermycin-gyrase B-induced dimerization. AB - Platelet-activating factor (PAF) is a phospholipid with potent and diverse physiological actions, particularly as a mediator of inflammation. We have reported previously that mutant G protein-coupled receptors (GPCRs) affect the functional properties of coexpressed wild-type human PAF receptor (hPAFR) (Le Gouill, C., Parent, J. L., Caron, C. A., Gaudreau, R., Volkov, L., Rola Pleszczynski, M., and Stankova, J. (1999) J. Biol. Chem. 274, 12548-12554). Increasing evidence suggests that dimerization of GPCRs may play an important role in the regulation of their biological activity. Additional data have also suggested that dimerization may be important in the subsequent internalization of the delta-opioid receptor. To investigate the specific role of dimerization in the internalization process of GPCRs, we generated a fusion protein of hPAFR and bacterial DNA gyrase B (GyrB), dimerized through the addition of coumermycin. We found that dimerization potentiates PAF-induced internalization of hPAFR-GyrB in Chinese hamster ovary cells stably expressing c-Myc-hPAFR-GyrB. Coumermycin driven dimerization was also sufficient to induce an agonist-independent sequestration process in an arrestin- and clathrin-independent manner. Moreover, the protein kinase C inhibitors staurosporine and GF109203X blocked the coumermycin-induced desensitization of hPAFR-GyrB, suggesting the implication of protein kinase C in the molecular mechanism mediating the agonist-independent desensitization of the receptor. Taken together, these findings suggest a novel mechanism of GPCR desensitization and internalization triggered by dimerization. PMID- 12756252 TI - Characterization of the metastasis-associated protein, S100A4. Roles of calcium binding and dimerization in cellular localization and interaction with myosin. AB - Elevated S100A4 protein expression is associated with metastatic tumor progression and appears to be a strong molecular marker for clinical prognosis. S100A4 is a calcium-binding protein that is known to form homodimers and interacts with several proteins in a calcium-dependent manner. Here we show that S100A4 localizes to lamellipodia structures in a migrating breast cancer-derived cell line and colocalizes with a known S100A4-interacting protein, myosin heavy chain IIA, at the leading edge. We demonstrate that S100A4 mutants that are defective in either their ability to dimerize or in calcium binding are unable to interact with myosin heavy chain IIA. An S100A4 mutant that is deficient for calcium binding retains the ability to form homodimers, suggesting that S100A4 can exist as calcium-free or calcium-bound dimers in vivo. However, a calcium bound S100A4 monomer only interacts with another calcium-bound monomer and not with an S100A4 mutant that does not bind calcium. Interestingly, despite the calcium dependence for interaction with known protein partners, calcium binding is not necessary for localization to lamellipodia. Both wild type and a mutant that is deficient for calcium binding colocalize with known markers of actively forming leading edges of lamellipodia, Arp3 and neuronal Wiskott-Aldrich syndrome protein. These data suggest that S100A4 localizes to the leading edge in a calcium-independent manner, and identification of the proteins that are involved in localizing S100A4 to the lamellipodial structures may provide novel insight into the mechanism by which S100A4 regulates metastasis. PMID- 12756253 TI - Structure of the bifunctional dCTP deaminase-dUTPase from Methanocaldococcus jannaschii and its relation to other homotrimeric dUTPases. AB - The bifunctional dCTP deaminase-dUTPase (DCD-DUT) from Methanocaldococcus jannaschii catalyzes the deamination of the cytosine moiety in dCTP and the hydrolysis of the triphosphate moiety forming dUMP, thereby preventing uracil from being incorporated into DNA. The crystal structure of DCD-DUT has been determined to 1.88-A resolution and represents the first known structure of an enzyme catalyzing dCTP deamination. The functional form of DCD-DUT is a homotrimer wherein the subunits are composed of a central distorted beta-barrel surrounded by two beta-sheets and four helices. The trimeric DCD-DUT shows structural similarity to trimeric dUTPases at the tertiary and quaternary levels. There are also additional structural elements in DCD-DUT compared with dUTPase because of a longer primary structure. Four of the five conserved sequence motifs that create the active sites in dUTPase are found in structurally equivalent positions in DCD-DUT. The last 25 C-terminal residues of the 204-residue-long DCD DUT are not visible in the electron density map, but, analogous to dUTPases, the C terminus is probably ordered, closing the active site upon catalysis. Unlike other enzymes catalyzing the deamination of cytosine compounds, DCD-DUT is not exploiting an enzyme-bound metal ion such as zinc or iron for nucleophile generation. The active site contains two water molecules that are engaged in hydrogen bonds to the invariant residues Ser118, Arg122, Thr130, and Glu145. These water molecules are potential nucleophile candidates in the deamination reaction. PMID- 12756254 TI - Recruitment of JNK to JIP1 and JNK-dependent JIP1 phosphorylation regulates JNK module dynamics and activation. AB - JIP1 is a scaffold protein that assembles and facilitates the activation of the mixed lineage kinase-dependent JNK module. Results of earlier work led us to propose a model for JIP1-JNK complex regulation that predicts that under basal conditions, JIP1 maintains DLK in a monomeric, unphosphorylated, and catalytically inactive state. Upon appropriate module stimulation, JNK-JIP1 binding affinity increases and DLK-JIP1 affinity decreases. Dissociation of DLK from JIP1 results in subsequent DLK oligomerization, autophosphorylation, and ultimately module activation. Our previous published results suggested the hypothesis that recruitment of JNK to JIP1 and phosphorylation of JIP1 by JNK is prerequisite for activation of the JNK module (Nihalani, D., Meyer, D., Pajni, S., and Holzman, L. B. (2001) EMBO J. 20, 3447-3458). The present study corroborated this hypothesis by demonstrating that JNK binding to JIP1 is necessary for stimulus-induced dissociation of DLK from JIP1, for DLK oligomerization, and for JNK activation. After mapping JNK-dependent JIP1 phosphorylation sites and testing their functional significance, it was observed that phosphorylation by JNK of JIP1 on Thr-103 and not other phosphorylated JIP1 residues is necessary for the regulation of DLK association with JIP1, DLK activation, and subsequent module activation. A refined model of JIP1-JNK module regulation is presented in which JNK phosphorylation of JIP1 is necessary prior to module activation. PMID- 12756255 TI - Requirement of domain-domain interaction for conformational change and functional ATP hydrolysis in myosin. AB - Coordination between the nucleotide-binding site and the converter domain of myosin is essential for its ATP-dependent motor activities. To unveil the communication pathway between these two sites, we investigated contact between side chains of Phe-482 in the relay helix and Gly-680 in the SH1-SH2 helix. F482A myosin, in which Phe-482 was changed to alanine with a smaller side chain, was not functional in vivo. In vitro, F482A myosin did not move actin filaments and the Mg2+-ATPase activity of F482A myosin was hardly activated by actin. Phosphate burst and tryptophan fluorescence analyses, as well as fluorescence resonance energy transfer measurements to estimate the movements of the lever arm domain, indicated that the transition from the open state to the closed state, which precedes ATP hydrolysis, is very slow. In contrast, F482A/G680F doubly mutated myosin was functional in vivo and in vitro. The fact that a larger side chain at the 680th position suppresses the defects of F482A myosin suggests that the defects are caused by insufficient contact between side chains of Ala-482 and Gly 680. Thus, the contact between these two side chains appears to play an important role in the coordinated conformational changes and subsequent ATP hydrolysis. PMID- 12756256 TI - Self-association of PAR-3-mediated by the conserved N-terminal domain contributes to the development of epithelial tight junctions. AB - PAR-3 is a scaffold-like PDZ-containing protein that forms a complex with PAR-6 and atypical protein kinase C (PAR-3-atypical protein kinase C-PAR-6 complex) and contributes to the establishment of cell polarity in a wide variety of biological contexts. In mammalian epithelial cells, it localizes to tight junctions, the most apical end of epithelial cell-cell junctions, and contributes to the formation of functional tight junctions. However, the mechanism by which PAR-3 localizes to tight junctions and contributes to their formation remains to be clarified. Here we show that the N-terminal conserved region, CR1-(1-86), and the sequence 937-1,024 are required for its recruitment to the most apical side of the cell-cell contact region in epithelial Madin-Darby canine kidney cells. We also show that CR1 self-associates to form an oligomeric complex in vivo and in vitro. Further, overexpression of CR1 in Madin-Darby canine kidney cells disturbs the distribution of atypical protein kinase C and PAR-6 as well as PAR-3 and delays the formation of functional tight junctions. These results support the notion that the CR1-mediated self-association of the PAR-3-containing protein complex plays a role during the formation of functional tight junctions. PMID- 12756257 TI - The unique properties of tonic smooth muscle emerge from intrinsic as well as intermolecular behaviors of Myosin molecules. AB - To better understand the molecular basis for some of the unique mechanical properties of tonic smooth muscle, we use a laser trap to assay the mechanochemistry of single smooth muscle heavy meromyosin molecules lacking a seven-amino acid insert in the nucleotide binding loop (minus insert). We measured a second-order ATP-induced actin dissociation rate, kT, of 2.2 x 10(6) m(-1) s(-1), an ADP release rate, k-D, of 19 s(-1), a second-order ADP binding rate, kD, of 60 x 10(5) m(-1) s(-1), and an ADP affinity, KD, of 3.2 microm, which is more than 100-fold greater than that measured for skeletal muscle myosin. By performing in vitro motility studies under nearly identical conditions, we show that the relatively slow actin velocity generated by minus insert heavy meromyosin is significantly influenced, but not limited, by k-D. Our results support a model in which two separate intermediate steps in the actin myosin catalyzed ATP hydrolysis reaction are energetically coupled through mechanical interactions, and we discuss this model in the context of the ability of tonic muscle to maintain high forces at low energetic cost (latch). PMID- 12756258 TI - Twitchin from molluscan catch muscle: primary structure and relationship between site-specific phosphorylation and mechanical function. AB - The phosphorylation state of the myosin thick filament-associated mini-titin, twitchin, regulates catch force maintenance in molluscan smooth muscle. The full length cDNA for twitchin from the anterior byssus retractor muscle of the mussel Mytilus was obtained using PCR and 5'rapid amplification of cDNA ends, and its derived amino acid sequence showed a large molecule ( approximately 530 kDa) with a motif arrangement as follows: (Ig)11(IgFn2)2Ig(Fn)3Ig(Fn)2Ig(Fn)3(Ig)2(Fn)2(Ig)2 FnKinase(Ig)4. Other regions of note include a 79-residue sequence between Ig domains 6 and 7 (from the N terminus) in which more than 60% of the residues are Pro, Glu, Val, or Lys and between the 7th and 8th Ig domains, a DFRXXL motif similar to that thought to be necessary for high affinity binding of myosin light chain kinase to F-actin. Two major phosphorylation sites, i.e. D1 and D2, were located in linker regions between Ig domains 7 and 8 and Ig domains 21 and 22, respectively. Correlation of the phosphorylation state of twitchin, using antibodies specific to D1 and D2, with mechanical properties suggested that phosphorylation of both D1 and D2 is required for relaxation from the catch state. PMID- 12756259 TI - Mismatch repair in human nuclear extracts: effects of internal DNA-hairpin structures between mismatches and excision-initiation nicks on mismatch correction and mismatch-provoked excision. AB - DNA mismatch repair (MMR) couples recognition of base mispairs by MSH2.MSH6 heterodimers to initiation, hundreds of nucleotides away, of nascent strand 3'-5' or 5'-3' excision through the mispair. Mismatch-recognition complexes have been hypothesized to move along DNA to excision-initiation signals, in eukaryotes, perhaps ends of nascent DNA, or to remain at mismatches and search through space for initiation signals. Subsequent MMR excision, whether simple processive digestion of the targeted strand or tracking of an excision complex, remains poorly understood. In human cell-free extracts, we analyzed correction of a mismatch in a 2.2-kilobase pair (kbp) circular plasmid containing a pre-existing excision-initiation nick for initiation, and measured MMR excision (in the absence of exogenous dNTPs) at specific locations. Excision specificities were approximately 100:1 for nicked versus continuous strands, 80:1 for mismatched versus homoduplex DNA, and 30:1 for shorter (0.3-kbp) versus longer (1.9-kbp) nick-mispair paths. To test models for recognition-excision coupling and excision progress, we inserted potential blockades, 20-bp hairpins, into nick-mispair paths, using a novel technique to first generate gapped plasmid. Continuous strand longer-path hairpins did not affect mismatch correction, but shorter-path hairpins reduced correction 4-fold, and both together eliminated it. Shorter-path hairpins had little effect on initiation of (3'-5') excision, measured 30-60 nucleotides 5' to the nick, but blocked subsequent progress of excision to the mismatch; longer-path hairpins blocked the (lower level) 5'-3' excision to the mismatch. Thus, (a) MMR excision protein(s) cannot move past DNA hairpins. Hairpins at both ends of substrate-derived 0.5-kbp DNA fragments did not prevent ATP-induced dissociation of mismatch-bound human MSH2.MSH6, so recognition complexes at mismatches might provoke excision at nicks beyond hairpins, or loosely sliding MSH2.MSH6 dimers might move to the nicks. PMID- 12756260 TI - Functional endothelin receptors are present on nuclei in cardiac ventricular myocytes. AB - Endothelins are thought to act through two specific, plasmalemmal G protein coupled receptor subtypes, ETAR and ETBR. However, in subfractionated cardiac membranes, ETAR immunoreactivity was detected only in the plasma membrane whereas ETBR immunoreactivity was detected predominantly in membranes of intracellular origin. Confocal microscopy demonstrated the presence of intracellular ETAR and ETBR in ventricular myocytes. ETAR were primarily on plasma membrane (surface membranes and transverse-tubules) and to a lesser extent on the nucleus while ETBR localized primarily to the nuclei. Western blot analysis of nuclei isolated from the heart indicated the presence of endothelin receptors: both ETAR and ETBR copurified with nucleoporin 62, whereas markers of endoplasmic reticulum and Golgi membranes were depleted. Radioligand binding studies revealed that isolated nuclei contain specific [125I]ET-1 binding sites. Specific [125I]ET-1 binding was reduced by 70-80% using the ETAR-selective antagonist BQ610 and 20-30% using the ETBR-specific antagonist BQ788. IRL-1620, an ETBR-specific agonist, also reduced [125I]ET-1 binding. Furthermore, ET-1 and IRL-1620 altered the incorporation of 32P into nuclear proteins and caused a transient increase in nuclear Ca2+ concentration. Hence, cardiac nuclei possess both ETAR and ETBR subtypes, which are functional with respect to ligand binding and are coupled to signaling mechanisms within the nuclear membrane. PMID- 12756261 TI - Global structural changes in annexin 12. The roles of phospholipid, Ca2+, and pH. AB - Ca2+-dependent membrane interaction has long been recognized as a general property of the annexin (ANX) family of proteins. More recently, it has become clear that ANXs can also undergo Ca2+-independent membrane interactions at mildly acidic pH. Here we use site-directed spin labeling in combination with circular dichroism and biochemical labeling methods to compare the structure and membrane topography of these two different membrane-bound forms of ANX12. Our results reveal strong similarities between the solution structure and the structure of the Ca2+-dependent membrane-bound form at neutral pH. In contrast, all Ca2+ independent membrane interactions tested resulted in large scale conformational changes and membrane insertion. Pairs of spin labels that were in close proximity across the interface of different domains of the protein in both the soluble and Ca2+-dependent membrane form were >25 A apart in the Ca2+-independent membrane bound form. Despite these major conformational changes, the overall secondary structure content did not appear to be strongly altered and ANX12 remained largely helical. Thus, Ca2+-independent membrane interaction leads to massive refolding but not unfolding. Refolding did not occur at low pH in the absence of membranes but occurred within a few seconds after phospholipid vesicles were added. The phospholipid composition of the vesicles was an important modulator of Ca2+-independent membrane interaction. For example, cardiolipin-containing vesicles induced Ca2+-independent membrane interaction even at near neutral pH, thereby raising the possibility that lipid composition could induce relatively rapid Ca2+-independent membrane interaction in vivo. PMID- 12756262 TI - Antigen-capturing cells can masquerade as memory B cells. AB - As well as classically defined switched immunoglobulin isotype-expressing B cells, memory B cells are now thought to include IgM-expressing cells and memory cells that lack B cell lineage markers, such as B220 or CD19. We set out to compare the relative importance of memory B cell subsets with an established flow cytometry method to identify antigen-specific cells. After immunization with PE, we could detect B220+ and, as reported previously, B220- antigen-binding cells (McHeyzer-Williams, L.J., M. Cool, and M.G. McHeyzer-Williams. 2001. J. Immunol. 167:1393-1405). The B220-PE+ cells bore few markers typical of B cells, but resembled myeloid cells. Further analysis of the antigen-binding characteristics of these cells showed that, upon immunization with two fluorescent proteins, the B220- cells could bind both. Furthermore, this subpopulation was detected in RAG1 /- mice after transfer of anti-PE mouse serum. These data strongly suggest that these cells capture serum Ig, via Fc receptors, and thus appear antigen-specific. Investigation of these antigen-capturing cells in a variety of knockout mice indicates that they bind monomeric IgG in an FcgammaR1 (CD64)-dependent manner. We find no evidence of a B220- memory B cell population that is not explicable by antigen-capturing cells, and warn that care must be taken when using antigen specificity or surface IgG as an indicator of B cell memory. PMID- 12756263 TI - NKG2D-mediated natural killer cell protection against cytomegalovirus is impaired by viral gp40 modulation of retinoic acid early inducible 1 gene molecules. AB - Natural killer (NK) cells play a critical role in the innate immune response against cytomegalovirus (CMV) infections. Although CMV encodes several gene products committed to evasion of adaptive immunity, viral modulation of NK cell activity is only beginning to be appreciated. A previous study demonstrated that the mouse CMV m152-encoded gp40 glycoprotein diminished expression of ligands for the activating NK cell receptor NKG2D on the surface of virus-infected cells. Here we have defined the precise ligands that are affected and have directly implicated NKG2D in immune responses to CMV infection in vitro and in vivo. Murine CMV (MCMV) infection potently induced transcription of all five known retinoic acid early inducible 1 (RAE-1) genes (RAE-1alpha, RAE-1beta, RAE-1delta, RAE-1 epsilon, and RAE-1gamma), but not H-60. gp40 specifically down-regulated the cell surface expression of all RAE-1 proteins, but not H-60, and diminished NK cell interferon gamma production against CMV-infected cells. Consistent with previous findings, a m152 deletion mutant virus (Deltam152) was less virulent in vivo than the wild-type Smith strain of MCMV. Treatment of BALB/c mice with a neutralizing anti-NKG2D antibody before infection increased titers of Deltam152 virus in the spleen and liver to levels seen with wild-type virus. These experiments demonstrate that gp40 impairs NK cell recognition of virus-infected cells through disrupting the RAE-1-NKG2D interaction. PMID- 12756264 TI - Lymphocyte homing to bronchus-associated lymphoid tissue (BALT) is mediated by L selectin/PNAd, alpha4beta1 integrin/VCAM-1, and LFA-1 adhesion pathways. AB - Bronchus-associated lymphoid tissue (BALT) participates in airway immune responses. However, little is known about the lymphocyte-endothelial adhesion cascades that recruit lymphocytes from blood into BALT. We show that high endothelial venules (HEVs) in BALT express substantial levels of VCAM-1, in marked contrast to HEVs in other secondary lymphoid tissues. BALT HEVs also express the L-selectin ligand PNAd. Anti-L-selectin, anti-PNAd, and anti-LFA-1 mAbs almost completely block the homing of B and T lymphocytes into BALT, whereas anti-alpha4 integrin and anti-VCAM-1 mAbs inhibit homing by nearly 40%. alpha4beta7 integrin and MAdCAM-1 are not involved. Importantly, we found that mAbs against alpha4 integrin and VCAM-1 significantly block the migration of total T cells (80% memory phenotype) but not naive T and B cells to BALT. These results suggest that an adhesion cascade, which includes L-selectin/PNAd, alpha4beta1 integrin/VCAM-1, and LFA-1, targets specific lymphocyte subsets to BALT. This high level of involvement of alpha4beta1 integrin/VCAM-1 is unique among secondary lymphoid tissues, and may help unify lymphocyte migration pathways and immune responses in BALT and other bronchopulmonary tissues. PMID- 12756265 TI - Steroid hormone synthesis by vaccinia virus suppresses the inflammatory response to infection. AB - The 3beta-hydroxysteroid dehydrogenase (3beta-HSD) isoenzymes play a key role in cellular steroid hormone synthesis. Vaccinia virus (VV) also synthesizes steroid hormones with a 3beta-HSD enzyme (v3beta-HSD) encoded by gene A44L. Here we examined the effects of v3beta-HSD in VV disease using wild-type (vA44L), deletion (vDeltaA44L), and revertant (vA44L-rev) viruses in a murine intranasal model. Loss of A44L was associated with an attenuated phenotype. Early (days 1-3) after infection with vDeltaA44L or control viruses the only difference observed between groups was the reduced corticosterone level in lungs and plasma of vDeltaA44L-infected animals. Other parameters examined (body weight, signs of illness, temperature, virus titres, the pulmonary inflammatory infiltrate, and interferon [IFN]-gamma levels) were indistinguishable between groups. Subsequently, vDeltaA44L-infected animals had reduced weight loss and signs of illness, and displayed a vigorous pulmonary inflammatory response. This was characterized by rapid recruitment of CD4+ and CD8+ lymphocytes, enhanced IFN gamma production and augmented cytotoxic T lymphocyte activity. These data suggest that steroid production by v3beta-HSD contributes to virus virulence by inhibiting an effective inflammatory response to infection. PMID- 12756266 TI - AID mediates hypermutation by deaminating single stranded DNA. AB - Activation-induced deaminase (AID) is a protein indispensable for the diversification of immunoglobulin (Ig) genes by somatic hypermutation (SHM), class switch recombination (CSR), and gene conversion. To date, the precise role of AID in these processes has not been determined. Here we demonstrate that purified, tetrameric AID can deaminate cytidine residues in DNA, but not in RNA. Furthermore, we show that AID will bind and deaminate only single-stranded DNA, which implies a direct, functional link between hypermutation and transcription. Finally, AID does not target mutational hotspots, thus mutational targeting to specific residues must be attributed to different factors. PMID- 12756267 TI - Enforced granulocyte/macrophage colony-stimulating factor signals do not support lymphopoiesis, but instruct lymphoid to myelomonocytic lineage conversion. AB - We evaluated the effects of ectopic granulocyte/macrophage colony-stimulating factor (GM-CSF) signals on hematopoietic commitment and differentiation. Lineage restricted progenitors purified from mice with the ubiquitous transgenic human GM CSF receptor (hGM-CSFR) were used for the analysis. In cultures with hGM-CSF alone, hGM-CSFR-expressing (hGM-CSFR+) granulocyte/monocyte progenitors (GMPs) and megakaryocyte/erythrocyte progenitors (MEPs) exclusively gave rise to granulocyte/monocyte (GM) and megakaryocyte/erythroid (MegE) colonies, respectively, providing formal proof that GM-CSF signals support the GM and MegE lineage differentiation without affecting the physiological myeloid fate. hGM CSFR transgenic mice were crossed with mice deficient in interleukin (IL)-7, an essential cytokine for T and B cell development. Administration of hGM-CSF in these mice could not restore T or B lymphopoiesis, indicating that enforced GM CSF signals cannot substitute for IL-7 to promote lymphopoiesis. Strikingly, >50% hGM-CSFR+ common lymphoid progenitors (CLPs) and >20% hGM-CSFR+ pro-T cells gave rise to granulocyte, monocyte, and/or myeloid dendritic cells, but not MegE lineage cells in the presence of hGM-CSF. Injection of hGM-CSF into mice transplanted with hGM-CSFR+ CLPs blocked their lymphoid differentiation, but induced development of GM cells in vivo. Thus, hGM-CSF transduces permissive signals for myeloerythroid differentiation, whereas it transmits potent instructive signals for the GM differentiation to CLPs and early T cell progenitors. These data suggest that a majority of CLPs and a fraction of pro-T cells possess plasticity for myelomonocytic differentiation that can be activated by ectopic GM-CSF signals, supporting the hypothesis that the down-regulation of GM-CSFR is a critical event in producing cells with a lymphoid-restricted lineage potential. PMID- 12756268 TI - Lysosomal membrane permeabilization induces cell death in a mitochondrion dependent fashion. AB - A number of diseases are due to lysosomal destabilization, which results in damaging cell loss. To investigate the mechanisms of lysosomal cell death, we characterized the cytotoxic action of two widely used quinolone antibiotics: ciprofloxacin (CPX) or norfloxacin (NFX). CPX or NFX plus UV light (NFX*) induce lysosomal membrane permeabilization (LMP), as detected by the release of cathepsins from lysosomes. Inhibition of the lysosomal accumulation of CPX or NFX suppresses their capacity to induce LMP and to kill cells. CPX- or NFX-triggered LMP results in caspase-independent cell death, with hallmarks of apoptosis such as chromatin condensation and phosphatidylserine exposure on the plasma membrane. LMP triggers mitochondrial membrane permeabilization (MMP), as detected by the release of cytochrome c. Both CPX and NFX* cause Bax and Bak to adopt their apoptotic conformation and to insert into mitochondrial membranes. Bax-/- Bak-/- double knockout cells fail to undergo MMP and cell death in response to CPX- or NFX-induced LMP. The single knockout of Bax or Bak (but not Bid) or the transfection-enforced expression of mitochondrion-targeted (but not endoplasmic reticulum-targeted) Bcl-2 conferred protection against CPX (but not NFX*)-induced MMP and death. Altogether, our data indicate that mitochondria are indispensable for cell death initiated by lysosomal destabilization. PMID- 12756269 TI - By altering ocular immune privilege, bone marrow-derived cells pathogenically contribute to DBA/2J pigmentary glaucoma. AB - Pigment dispersion syndrome causes iris pigment release and often progresses to elevated intraocular pressure and pigmentary glaucoma (PG). Because melanin pigment can have adjuvant like properties and because the Gpnmb gene, which contributes to pigment dispersion in DBA/2J (D2) mice, is expressed in dendritic cells, we tested the hypothesis that ocular immune abnormalities participate in PG phenotypes. Strikingly, we show that D2 eyes exhibit defects of the normally immunosuppressive ocular microenvironment including inability of aqueous humor to inhibit T cell activation, failure to support anterior chamber (AC)-associated immune deviation, and loss of ocular immune privilege. Histologic analysis demonstrates infiltration of inflammatory leukocytes into the AC and their accumulation within the iris, whereas clinical indications of inflammation are typically very mild to undetectable. Importantly, some of these abnormalities precede clinical indications of pigment dispersal, suggesting an early role in disease etiology. Using bone marrow chimeras, we show that lymphohematopoietic cell lineages largely dictate the progression of pigment dispersion, the ability of the eye to support induction of AC-associated immune deviation, and the integrity of the blood/ocular barrier. These results suggest previously unsuspected roles for bone marrow-derived cells and ocular immune privilege in the pathogenesis of PG. PMID- 12756270 TI - A new type of metal recognition by human T cells: contact residues for peptide independent bridging of T cell receptor and major histocompatibility complex by nickel. AB - In spite of high frequencies of metal allergies, the structural basis for major histocompatibility complex (MHC)-restricted metal recognition is among the unanswered questions in the field of T cell activation. For the human T cell clone SE9, we have identified potential Ni contact sites in the T cell receptor (TCR) and the restricting human histocompatibility leukocyte antigen (HLA)-DR structure. The specificity of this HLA-DR-promiscuous VA22/VB17+ TCR is primarily harbored in its alpha chain. Ni reactivity is neither dependent on protein processing in antigen-presenting cells nor affected by the nature of HLA-DR associated peptides. However, SE9 activation by Ni crucially depends on Tyr29 in CDR1alpha, an N-nucleotide-encoded Tyr94 in CDR3alpha, and a conserved His81 in the HLA-DR beta chain. These data indicate that labile, nonactivating complexes between the SE9 TCR and most HLA-DR/peptide conjugates might supply sterically optimized coordination sites for Ni ions, three of which were identified in this study. In such complexes Ni may effectively bridge the TCR alpha chain to His81 of most DR molecules. Thus, in analogy to superantigens, Ni may directly link TCR and MHC in a peptide-independent manner. However, unlike superantigens, Ni requires idiotypic, i.e., CDR3alpha-determined TCR amino acids. This new type of TCR-MHC linkage might explain the high frequency of Ni-reactive T cells in the human population. PMID- 12756271 TI - P-selectin glycoprotein ligand-1 mediates L-selectin-dependent leukocyte rolling in venules. AB - Leukocyte rolling in postcapillary venules of inflamed tissues is reduced in L selectin-deficient mice and mice treated with L-selectin blocking antibodies, but the glycoprotein ligand for L-selectin in inflamed venules is unknown. Here, we show that L-selectin-dependent rolling after P-selectin blockade is completely absent in P-selectin glycoprotein ligand-1 (PSGL-1)-/- mice or wild-type mice treated with a PSGL-1 blocking monoclonal antibody. Immunohistochemistry and flow cytometry failed to show PSGL-1 expression on resting or inflamed endothelium or on platelets. To investigate whether leukocyte-expressed PSGL-1 is mediating L selectin-dependent rolling, we reconstituted lethally irradiated wild-type mice with PSGL-1-/- bone marrow cells. These chimeric mice showed no L-selectin dependent rolling, suggesting that leukocyte-expressed PSGL-1 mediates L-selectin dependent rolling. Frame-to-frame video analysis of L-selectin-dependent rolling in wild-type mice showed that the majority of observed L-selectin-dependent leukocyte rolling was between free flowing leukocytes and already adherent leukocytes or possibly leukocyte fragments, followed by E-selectin-dependent leukocyte rolling along the endothelium. Leukocyte rolling was significantly slower for leukocyte-endothelial than leukocyte-leukocyte interactions. We conclude that leukocyte-expressed PSGL-1 serves as the main L-selectin ligand in inflamed postcapillary venules. L-selectin binding to PSGL-1 initiates tethering events that enable L-selectin-independent leukocyte-endothelial interactions. These findings provide a molecular mechanism for the inflammatory defects seen in L-selectin-deficient mice. PMID- 12756273 TI - Role of Na(+), K(+)-ATPase in morphine-induced antinociception. AB - We evaluated the modulation by Na+,K+-ATPase inhibitors of morphine-induced antinociception in the tail-flick test and [3H]naloxone binding to forebrain membranes. The antinociception induced by morphine (1-32 mg/kg, s.c.) in mice was dose-dependently antagonized by ouabain (1-10 ng/mouse, i.c.v.), which produced a significant shift to the right of the morphine dose-response curve. The i.c.v. administration of three Na+,K+-ATPase inhibitors (ouabain at 0.1-100, digoxin at 1-1000, and digitoxin at 10-10000 ng/mouse) dose-dependently antagonized the antinociceptive effect of morphine (4 mg/kg, s.c.) in mice, with the following order of potency: ouabain > digoxin > digitoxin. This effect cannot be explained by any interaction at opioid receptors, since none of these Na+,K+-ATPase inhibitors displaced [3H]naloxone from its binding sites, whereas naloxone did so in a concentration-dependent manner. The antinociception induced by morphine (5 mg/kg, s.c.) in rats was antagonized by the i.c.v. administration of ouabain at 10 ng/rat, whereas it was not significantly modified by intrathecally administered ouabain (10 and 100 ng/rat). These results suggest that the activation of Na+,K+-ATPase plays a role in the supraspinal, but not spinal, antinociceptive effect of morphine. PMID- 12756274 TI - The role of I(1)-imidazoline and alpha(2)-adrenergic receptors in the modulation of glucose metabolism in the spontaneously hypertensive obese rat model of metabolic syndrome X. AB - We examined glucose metabolism after I1-imidazoline (I1R) and alpha2-adrenergic receptor (alpha2AR) activation in an animal model of metabolic syndrome X. Fasted spontaneously hypertensive obese rats (SHROB) were given the I1R/alpha2AR agonists moxonidine and rilmenidine or the alpha2AR agonist guanabenz. Because of the dual specificity of moxonidine, its actions were split into adrenergic and nonadrenergic components by using selective antagonists: rauwolscine (alpha2AR) efaroxan (I1R/alpha2AR), or 2-endo-amino-3-exo-isopropylbicyclo[2.2.1.]heptane (AGN 192403) (I1R). Hyperglycemia induced by moxonidine, rilmenidine, and guanabenz resulted from inhibition of insulin secretion. Similar responses were observed after oral dosing and in lean littermates. Glucagon was reduced by the I1R agonists (moxonidine, 32 +/- 5%; rilmenidine, 24 +/- 7%) but elevated by guanabenz (71 +/- 32%). The hyperglycemic and hypoinsulinemic responses to moxonidine were blocked by rauwolscine. In contrast, rauwolscine potentiated the reduction in glucagon (39 +/- 6%). AGN 193402 blocked the glucagon response without affecting hyperglycemia and hypoinsulinemia. Efaroxan blocked all responses to moxonidine. When SHROB rats were treated with moxonidine 15 min before an oral glucose tolerance test, the glucose area under the curve (AUC) was increased. Antagonizing the alpha2AR component of moxonidine's action with rauwolscine improved glucose AUC 3-fold and facilitated the insulin secretory response and reduced glucagon secretion. Testing fasting glucose and insulin during 3 weeks of oral moxonidine revealed early hyperglycemia that later faded, and a progressive drop in fasting insulin. The acute hyperglycemia and hypoinsulinemia elicited by moxonidine and rilmenidine was mediated by alpha2AR, whereas I1R may reduce glucagon and increase insulin, particularly after a glucose load. PMID- 12756272 TI - Competition between two MHC binding registers in a single peptide processed from myelin basic protein influences tolerance and susceptibility to autoimmunity. AB - Experimental allergic encephalomyelitis (EAE) is an animal model for multiple sclerosis induced by stimulating myelin basic protein (MBP)-specific T cells. The MBP-specific repertoire in B10.PL mice is shaped by tolerance mechanisms that eliminate MBP121-150-specific T cells. In contrast, MBPAc1-11-specific T cells escape tolerance and constitute the encephalitogenic repertoire. To determine if this differential tolerance is caused by differences in the abundance of MBP epitopes generated by processing, MBP peptides were eluted from I-Au complexes and analyzed by mass spectrometry. Peptides were identified from both the NH2 terminal and MBP121-150 regions. Unexpectedly, MBPAc1-18 and Ac1-17, which contain the MBPAc1-11 epitope, were much more abundant than MBP121-150 peptides. The results demonstrate that competition between two I-Au binding registers, a low affinity register defined by MBPAc1-11 and a high affinity register defined by MBP5-16, prevents most of the NH2-terminal naturally processed peptides from binding in the MBPAc1-11 register. The small fraction of MBPAc1-18 bound in the MBPAc1-11 register is not sufficient to induce tolerance but provides a ligand for MBPAc1-11-specific T cells during disease. These results provide a basis for both the lack of tolerance to MBPAc1-11 and the ability of this epitope to become a target during autoimmunity. PMID- 12756276 TI - Diffusion, perfusion and the exclusion principles in the structural and functional organization of the living cell: reappraisal of the properties of the 'ground substance'. AB - The thesis is presented that only within very small microdomains of the cell internum might diffusion operate in the sorting of molecular affinities. Much of cell metabolism is guided and controlled in rate by the speed with which molecules that have to interact encounter one another. What is clear, however, is that the cell does not have a single 'modus operandi' but has the choice of many different strategies, each of which can contribute in different proportion to the rate of ongoing activity. It is probably our own desire to simplify things and use the most (or more) probable strategy that confines our appreciation of the overall robustness of the cell as a 'survival machine'. The main operative process at any given time (perfusion, diffusion or whatever) has always to be considered very carefully in relation to the organisational structure of the cell, which can be transient and fickle but nevertheless has been seen as involving an extensive cytomatrix, a ground substance, within an aqueous environment in which the degree of water structuring is even more fickle. PMID- 12756277 TI - Actin-based photo-orientation movement of chloroplasts in plant cells. AB - In photosynthesizing plant cells, chloroplasts change their arrangements and/or positions in response to light irradiation. These photo-orientation movements of chloroplasts are believed to play important roles in optimizing the photosynthetic activity of plant cells. We have been investigating the roles of the actin cytoskeleton in the intracellular movement and positioning of chloroplasts using the aquatic monocot Vallisneria gigantea Graebner and the terrestrial dicot Spinacia oleracea L. (spinach). In Vallisneria epidermal cells, chloroplasts accumulate on the cytoplasmic layer facing the top surface (outer periclinal layer) under dim red light, whereas they move to the cytoplasmic layer perpendicular to the outer periclinal layer (anticlinal layer) under strong blue light. Concomitant with these responses, actin filaments exhibit dramatic changes in their configurations. The possible modes of action of the actin cytoskeleton to regulate the movement and positioning of chloroplasts are briefly summarized, together with our recent analysis of the association of actin filaments with chloroplasts isolated from spinach leaves. PMID- 12756278 TI - Force-velocity relationships in actin-myosin interactions causing cytoplasmic streaming in algal cells. AB - Cytoplasmic streaming in giant internodal cells of green algae is caused by ATP dependent sliding between actin cables fixed on chloroplast rows and cytoplasmic myosin molecules attached to cytoplasmic organelles. Its velocity (>/=50 micro m s(-1)) is many times larger than the maximum velocity of actin-myosin sliding in muscle. We studied kinetic properties of actin-myosin sliding causing cytoplasmic streaming in internodal cell preparations of Chara corallina, into which polystyrene beads, coated with cytoplasmic myosin molecules, were introduced. Constant centrifugal forces directed opposite to the bead movement were applied as external loads. The steady-state force-velocity (P-V) curves obtained were nearly straight, irrespective of the maximum isometric force generated by cytoplasmic myosin molecules, indicating a large duty ratio of cytoplasmic myosin head. The large velocity of cytoplasmic streaming can be accounted for, at least qualitatively, by assuming a mechanically coupled interaction between cytoplasmic myosin heads as well as a large distance of unitary actin-myosin sliding. PMID- 12756279 TI - Actin comet tails, endosomes and endosymbionts. AB - The Arp2/3 complex consists of seven highly conserved and tightly associated subunits, two of which are the actin-related proteins Arp2 and Arp3. One of the best-studied functions of the Arp2/3 complex is to stimulate actin nucleation and force production at the leading edge of motile cells. What is now clear is that Arp2/3-complex-mediated force production drives many intracellular movements, including movement of bacterial pathogens in infected host cells, internalization of extracellular materials via phagocytosis and endocytosis, and movement of mitochondria during cell division in budding yeast. Here, we describe recent advances in the mechanisms underlying Arp2/3 complex-driven intracellular movement. PMID- 12756281 TI - Mitochondria are morphologically heterogeneous within cells. AB - Mitochondria play key roles in the life and death of cells. We investigated whether mitochondria represent morphologically continuous entities within single intact cells. Physical continuity of mitochondria was determined by three dimensional reconstruction of fluorescence from mitochondrially targeted DsRed1 or tetra-methyl rhodamine ethyl ester (TMRE). The mitochondria of pancreatic acinar, porcine aortic endothelial (PAE) cells, COS-7 cells and SH-SY5Y cells and neocortical astrocytes all displayed heterogeneous distributions and were of varying sizes. In general, there was a denser aggregation of mitochondria in perinuclear positions than in the cell periphery, where individual isolated mitochondria could clearly be seen. DsRed1 was found to be highly mobile within the matrix of individual mitochondria, with an estimated linear diffusion rate of 1 micro m s(-1). High-intensity irradiation of subcellular regions bleached the fluorescence of mitochondrially targeted DsRed1, but did not cause the mitochondria to depolarise or fragment. A lack of rapid fluorescence-recovery after-photobleaching (FRAP) of DsRed1 indicated lumenal discontinuity between mitochondria. We observed a slow (half-time approx. 20 min) recovery of DsRed1 fluorescence within the irradiated area that was attributed to mitochondrial movement or fusion of unbleached and bleached organelles. Mitochondria were not electrically coupled, since typically only individual mitochondria were observed to depolarise following irradiation of TMRE-loaded cells. Our data indicate that the mitochondria within individual cells are morphologically heterogeneous and unconnected, thus allowing them to have distinct functional properties. PMID- 12756280 TI - Mitochondrial movement and positioning in axons: the role of growth factor signaling. AB - The extreme length of axonal processes requires that aerobic ATP production and Ca(2+) homeostasis are non-uniformly organized in the cytoplasm. As a result, the transport and positioning of mitochondria along axons is essential for neuronal homeostasis. Mitochondria undergo rapid but intermittent transport in both the anterograde and retrograde directions in axons. We have shown that in chick embryonic sensory neurons, the transport of mitochondria responds to physiological changes in the cell and, particularly, to growth cone activity. When an axon is actively elongating, mitochondria move preferentially anterograde and then become stationary, accumulating in the region of the active growth cone. When axonal elongation ceases, mitochondria in the distal axon resume movement but undergo net retrograde transport and become uniformly distributed along the axon. This redistribution of mitochondria is achieved in two ways: there is a transition between motile and stationary mitochondria and a large up- and downregulation of their anterograde, but not retrograde, motor activity. Mitochondrial transport does not respond to the experimentally induced elongation of axons in the absence of an active growth cone, implying that signals from the active growth cone regulate transport. To determine the nature of these signals, we have focally stimulated the shafts of sensory axons in culture with nerve growth factor (NGF) covalently conjugated to polystyrene beads. We find that mitochondria accumulate at regions of focal NGF stimulation. This response is specific to mitochondria and does not result from general disruption of the cytoskeleton in the region of stimulation. Disruption of the phosphoinositide 3 kinase (PI 3-kinase) pathway, one of the signaling pathways downstream from NGF receptor binding, completely eliminates NGF effects on mitochondrial behavior in axons. We propose that mitochondrial transport and/or docking are regulated in part via NGF/TrkA/PI 3-kinase signaling. PMID- 12756282 TI - Intracellular convection, homeostasis and metabolic regulation. AB - Two views currently dominate experimental approaches to metabolic regulation. The first, let us call it Model 1, assumes that cells behave like a watery bag of enzymes. The alternative Model 2, however, assumes that 3-dimensional order and structure constrain metabolite behavior. A major problem in cell metabolism is determining why essentially all metabolite concentrations are remarkably stable (homeostatic) over large changes in pathway fluxes-for convenience, this is termed the [s] stability paradox. During large-scale transitions from maintenance metabolic rates to maximally activated work, contrasting demands of intracellular homeostasis versus metabolic regulation obviously arise. Data accumulated over the last 3-4 decades now make it clear that the demands of homeostasis prevail: during rest-work transitions, metabolites such as ATP and O(2) are notably and rigorously homeostatic; other intermediates usually do not vary by more than 0.5- to threefold over the resting condition. This impressive homeostasis is maintained despite changes in pathway fluxes that can exceed two orders of magnitude. Classical or Model 1 approaches to this problem can explain metabolite homeostasis, but the mechanisms for each metabolite, each enzyme locus, are necessarily specific. Thus Model 1 approaches basically do not provide a global explanation for the [s] stability paradox. Model 2 takes a different tack and assumes that an intracellular convection system acts as an over-riding 'assist' mechanism for facilitating enzyme-substrate encounter. Model 2 postulates that intracellular movement and convection are powered by macromolecular motors (unconventional myosins, dyneins, kinesin) running on actin or tubulin tracks. For fast and slow muscle fibers, microfilaments are concentrated near the periphery (where convection may be most important), but also extend throughout the actomyosin contractile apparatus both in horizontal and vertical dimensions. To this point in the development of the field, Model 1 and Model 2 approaches have operated as 'two solitudes', each considering the other incompatible with its own experimental modus operandi. In order to finally assemble a model that can sensibly explain a realistic working range of metabolic systems, opening of channels of communication between the above two very differing views of metabolic regulation would seem to be the requirement for the future. PMID- 12756283 TI - Myoglobin function reassessed. AB - The heart and those striated muscles that contract for long periods, having available almost limitless oxygen, operate in sustained steady states of low sarcoplasmic oxygen pressure that resist change in response to changing muscle work or oxygen supply. Most of the oxygen pressure drop from the erythrocyte to the mitochondrion occurs across the capillary wall. Within the sarcoplasm, myoglobin, a mobile carrier of oxygen, is developed in response to mitochondrial demand and augments the flow of oxygen to the mitochondria. Myoglobin-facilitated oxygen diffusion, perhaps by virtue of reduction of dimensionality of diffusion from three dimensions towards two dimensions in the narrow spaces available between mitochondria, is rapid relative to other parameters of cell respiration. Consequently, intracellular gradients of oxygen pressure are shallow, and sarcoplasmic oxygen pressure is nearly the same everywhere. Sarcoplasmic oxygen pressure, buffered near 0.33 kPa (2.5 torr; equivalent to approximately 4 micro mol l(-1) oxygen) by equilibrium with myoglobin, falls close to the operational K(m) of cytochrome oxidase for oxygen, and any small increment in sarcoplasmic oxygen pressure will be countered by increased oxygen utilization. The concentration of nitric oxide within the myocyte results from a balance of endogenous synthesis and removal by oxymyoglobin-catalyzed dioxygenation to the innocuous nitrate. Oxymyoglobin, by controlling sarcoplasmic nitric oxide concentration, helps assure the steady state in which inflow of oxygen into the myocyte equals the rate of oxygen consumption. PMID- 12756284 TI - Shaken and stirred: muscle structure and metabolism. AB - Muscles are ideal models with which to examine the relationship between structure and metabolism because they are some of the most highly structured cells, and are capable of the largest and most rapid metabolic transitions as well as the highest metabolic rates known. Studies of metabolism have traditionally been conducted within what can considered as the kinetic paradigm provided by 'solution biochemistry'; i.e. the rates of enzymatic reactions are studied in terms of their regulation by mass-action and allosteric effectors and, most recently, metabolic control analysis of pathways. This approach has served biology well and continues to be useful. Here, we consider the diffusion of small and large molecules in muscles and energy metabolism in the context of intracellular space. We find that in attempting to explain certain phenomena, a purely kinetic paradigm appears insufficient. Instead, phenomena such as the 'shuttling' of high-energy phosphate donors and acceptors and the binding of metabolic enzymes to intracellular structures or to each other are better understood when metabolic rates and their regulation are considered in the context of intracellular compartments, distances, gradients and diffusion. As in all of biology, however, complexity dominates, and to such a degree that one pathway may consist of several reactions that each behave according to different rules. 'Soluble' creatine kinase operates at or near equilibrium, while mitochondrial and myofibrillar creatine kinases directly channel substrate to (or from) the adenine nucleotide translocase and actomyosin-ATPase, their operation being thus displaced from equilibrium. Hexose 6-phosphate metabolism appears to obey the rules of solution biochemistry, e.g. phosphoglucoisomerase behaves as Haldane would have predicted in 1930. In contrast, given low steady-state substrate and product concentrations and high flux rates, a number of glycolytic reactions further downstream must be catalyzed by enzymes localized in close proximity to each other. Metabolites may be channeled within these complexes. When observed, mechanistic differences between species in the same steps or processes should not be surprising, considering how animals vary so much in structures, mechanical properties, mitochondrial contents and metabolic rates. This analysis suggests that declarations of the triumph of one mechanism or paradigm over all others, as well as calls for the abandonment of solution biochemistry, are unwarranted. Rather, metabolic biochemistry would seem better served by reconciling the old and the new. PMID- 12756285 TI - Analysis of glycolytic enzyme co-localization in Drosophila flight muscle. AB - In Drosophila flight muscles, glycolytic enzymes are co-localized along sarcomeres at M-lines and Z-discs and co-localization is required for normal flight. We have extended our analysis of this phenomenon to include a set of six glycolytic enzymes that catalyze consecutive reactions along the glycolytic pathway: aldolase, glycerol-3-phosphate dehydrogenase (GPDH), glyceraldehyde-3 phosphate dehydrogenase (GAPDH), triose phosphate isomerase, phosphoglycerate kinase and phosphoglycerol mutase (PGLYM). Each of these enzymes has an identical pattern of localization. In mutants null for GPDH, localization of none of the other enzymes occurs and therefore is interdependent. In optimally fixed preparations of myofibrils, accumulation of the enzymes at M-lines is much greater than at Z-discs. However, localization at M-lines is more labile, as shown by loss of localization when fixation is delayed. We have begun to analyze the protein-protein interaction involved in glycolytic enzyme co-localization using the yeast two-hybrid system. We have identified two pair-wise interactions. One is between GPDH and GAPDH and another is between GPDH and PGLYM. PMID- 12756286 TI - Phosphotransfer networks and cellular energetics. AB - Precise coupling of spatially separated intracellular ATP-producing and ATP consuming processes is fundamental to the bioenergetics of living organisms, ensuring a fail-safe operation of the energetic system over a broad range of cellular functional activities. Here, we provide an overview of the role of spatially arranged enzymatic networks, catalyzed by creatine kinase, adenylate kinase, carbonic anhydrase and glycolytic enzymes, in efficient high-energy phosphoryl transfer and signal communication in the cell. Studies of transgenic creatine kinase and adenylate kinase deficient mice, along with pharmacological targeting of individual enzymes, have revealed the importance of near-equilibrium reactions in the dissipation of metabolite gradients and communication of energetic signals to distinct intracellular compartments, including the cell nucleus and membrane metabolic sensors. Enzymatic capacities, isoform distribution and the dynamics of net phosphoryl flux through the integrated phosphotransfer systems tightly correlate with cellular functions, indicating a critical role of such networks in efficient energy transfer and distribution, thereby securing the cellular economy and energetic homeostasis under stress. PMID- 12756287 TI - Isozymes of mammalian hexokinase: structure, subcellular localization and metabolic function. AB - The first step in metabolism of glucose (Glc) is usually phosphorylation, catalyzed by hexokinase. However, the Glc-6-P produced can then enter one or more of several alternative pathways. Selective expression of isozymic forms of hexokinase, differing in catalytic and regulatory properties as well as subcellular localization, is likely to be an important factor in determining the pattern of Glc metabolism in mammalian tissues/cells. Despite their overall structural similarity, the Type I, Type II and Type III isozymes differ in important respects. All three isozymes are inhibited by the product, Glc-6-P, but with the Type I isozyme, this inhibition is antagonized by P(I), whereas with the Type II and Type III isozymes, P(i) actually causes additional inhibition. Reciprocal changes in intracellular levels of Glc-6-P and P(i) are closely associated with cellular energy status, and it is proposed that the response of the Type I isozyme to these effectors adapts it for catabolic function, introducing Glc into glycolytic metabolism for energy production. In contrast, the Type II, and probably the Type III, isozymes are suggested to serve primarily anabolic functions, e.g. to provide Glc-6-P for glycogen synthesis or metabolism via the pentose phosphate pathway for lipid synthesis. Type I hexokinase binds to mitochondria through interaction with porin, the protein that forms channels through which metabolites traverse the outer mitochondrial membrane. Several experimental approaches have led to the conclusion that the Type I isozyme, bound to actively phosphorylating mitochondria, selectively uses intramitochondrial ATP as substrate. Such interactions are thought to facilitate coordination of the introduction of Glc into glycolysis, via the hexokinase reaction, with the terminal oxidative stages of Glc metabolism occurring in the mitochondria, thus ensuring an overall rate of Glc metabolism commensurate with cellular energy demands and avoiding excessive production of lactate. The Type II isozyme also binds to mitochondria. Whether such coupling occurs with mitochondrially bound Type II hexokinase in normal tissues, and how it might be related to the proposed anabolic role of this isozyme, remain to be determined. The Type III isozyme lacks the hydrophobic N-terminal sequence known to be critical for binding of the Type I and Type II isozymes to mitochondria. Immunolocalization studies have indicated that, in many cell types, the Type III has a perinuclear localization, the possible metabolic consequences of which remain unclear. PMID- 12756288 TI - Metabolic consequences of functional complexes of mitochondria, myofibrils and sarcoplasmic reticulum in muscle cells. AB - Regulation of mitochondrial respiration both by endogenous and exogenous ADP in the cells in situ was studied in isolated and permeabilized cardiomyocytes, permeabilized cardiac fibers and 'ghost' fibers (all with a diameter of 10-20 micro m) at different (0-3 micro moll(-1)) free Ca(2+) concentrations in the medium. In all these preparations, the apparent K(m) of mitochondrial respiration for exogenous ADP at free Ca(2+) concentrations of 0-0.1 micro moll(-1) was very high, in the range of 250-350 micro moll(-1), in contrast to isolated mitochondria in vitro (apparent K(m) for ADP is approximately 20 micro moll(-1)). An increase in the free Ca(2+) concentration (up to 3 micro moll(-1), which is within physiological range), resulted in a very significant decrease of the apparent K(m) value to 20-30 micro moll(-1), a decrease of V(max) of respiration in permeabilized intact fibers and a strong contraction of sarcomeres. In ghost cardiac fibers, from which myosin was extracted but mitochondria were intact, neither the high apparent K(m) for ADP (300-350 micro moll(-1)) nor V(max) of respiration changed in the range of free Ca(2+) concentration studied, and no sarcomere contraction was observed. The exogenous-ADP-trapping system (pyruvate kinase + phosphoenolpyruvate) inhibited endogenous-ADP-supported respiration in permeabilized cells by no more than 40%, and this inhibition was reversed by creatine due to activation of mitochondrial creatine kinase. These results are taken to show strong structural associations (functional complexes) among mitochondria, sarcomeres and sarcoplasmic reticulum. Inside these complexes, mitochondrial functional state is controlled by channeling of ADP, mostly via energy- and phosphoryl-transfer networks, and apparently depends on the state of sarcomere structures. PMID- 12756289 TI - Four-dimensional organization of protein kinase signaling cascades: the roles of diffusion, endocytosis and molecular motors. AB - Extracellular signals received by membrane receptors are processed, encoded and transferred to the nucleus via phosphorylation and spatial relocation of protein members of multiple component pathways, such as mitogen activated protein kinase (MAPK) cascades. The receptor-induced membrane recruitment of the cytoplasmic protein SOS results in the activation of the Ras/MAPK cascade. It has been suggested that the membrane recruitment of signaling proteins causes an increase in the diffusion-limited rates. We have recently shown that this increase is too small to be responsible for enhanced signal transduction. Instead we demonstrate that the function of membrane localization is to increase the number (or average lifetime) of complexes between signaling partners. A hallmark of signaling pathways is the spatial separation of activation and deactivation mechanisms; e.g. a protein can be phosphorylated at the cell surface by a membrane-bound kinase and dephosphorylated in the cytosol by a cytosolic phosphatase. Given the measured values of protein diffusion coefficients and of phosphatase and kinase activities, the spatial separation is shown to result in precipitous phospho protein gradients. When information transfer is hampered by slow protein diffusion and rapid dephosphorylation, phospho-protein trafficking within endocytic vesicles may be an efficient way to deliver messages to physiologically relevant locations. The proposed mechanism explains recent observations that various inhibitors of endocytosis can inhibit MAPK activation. Additional mechanisms facilitating the relay of signals from cell-surface receptors to the nucleus can involve the assembly of protein kinases on a scaffolding protein and active transport of signaling complexes by molecular motors. We also discuss long range signaling within a cell, such as survival signaling in neurons. We hypothesize that ligand-independent waves of receptor activation or/and traveling waves of phosphorylated kinases emerge to spread the signals over long distances. PMID- 12756290 TI - The caveolar nitric oxide synthase/arginine regeneration system for NO production in endothelial cells. AB - The enzyme endothelial nitric oxide synthase (eNOS) catalyzes the conversion of arginine, oxygen and NADPH to NO and citrulline. Previous results suggest an efficient, compartmentalized system for recycling of citrulline to arginine utilized for NO production. In support of this hypothesis, the recycling enzymes, argininosuccinate synthase (AS) and argininosuccinate lyase (AL), have been shown to colocalize with eNOS in caveolae, a subcompartment of the plasma membrane. Under unstimulated conditions, the degree of recycling is minimal. Upon stimulation of NO production by bradykinin, however, recycling is co-stimulated to the extent that more than 80% of the citrulline produced is recycled to arginine. These results suggest an efficient caveolar recycling complex that supports the receptor-mediated stimulation of endothelial NO production. To investigate the molecular basis for the unique location and function of endothelial AS and AL, endothelial AS mRNA was compared with liver AS mRNA. No differences were found in the coding region of the mRNA species, but significant differences were found in the 5'-untranslated region (5'-UTR). The results of these studies suggest that sequence in the endothelial AS-encoding gene, represented by position -92 nt to -43 nt from the translation start site in the extended AS mRNA 5'-UTRs, plays an important role in differential and tissue specific expression. Overall, a strong evidential case has been developed supporting the proposal that arginine availability, governed by a caveolar localized arginine regeneration system, plays a key role in receptor-mediated endothelial NO production. PMID- 12756291 TI - Tight-adherence genes of Actinobacillus actinomycetemcomitans are required for virulence in a rat model. AB - Actinobacillus actinomycetemcomitans is a Gram-negative coccobacillus that has been associated with localized aggressive periodontitis and infections of the heart, brain, and urinary tract. Wild-type clinical isolates have the remarkable ability to adhere tenaciously and nonspecifically to solid surfaces such as glass, plastic, and hydroxyapatite. Adherence by A. actinomycetemcomitans is mediated by the tight-adherence (tad) gene locus, which consists of 14 genes (flp 1-flp-2-tadV-rcpCAB-tadZABCDEFG). All but 2 of the genes have been shown to be required for the secretion and assembly of long, bundled Flp1 fibrils. To test whether the tad locus is required for colonization and disease, we developed a rat model for periodontal disease. To mimic the natural route of infection, Sprague-Dawley rats were inoculated orally by adding bacteria directly to their food for 8 days. After inoculation with wild-type or mutant strains defective in adherence (flp-1 and tadA), the rats were assessed for colonization of the oral cavity and pathogenesis. Wild-type A. actinomycetemcomitans was able to colonize and persist for at least 12 weeks in the oral cavity, elicit a humoral immune response, and cause significant bone loss in rats. In contrast, rats fed flp-1 or tadA mutant strains showed no bone loss and their immune responses were indistinguishable from those of the uninoculated controls. These results demonstrate the critical importance of the tad locus in the colonization and pathogenesis of A. actinomycetemcomitans. PMID- 12756292 TI - Suppression of macrophage inflammatory responses by PPARs. PMID- 12756293 TI - Myocardin is a master regulator of smooth muscle gene expression. AB - Virtually all smooth muscle genes analyzed to date contain two or more essential binding sites for serum response factor (SRF) in their control regions. Because SRF is expressed in a wide range of cell types, it alone cannot account for smooth muscle-specific gene expression. We show that myocardin, a cardiac muscle- and smooth muscle-specific transcriptional coactivator of SRF, can activate smooth muscle gene expression in a variety of nonmuscle cell types via its association with SRF. Homodimerization of myocardin is required for maximal transcriptional activity and provides a mechanism for cooperative activation of smooth muscle genes by SRF-myocardin complexes bound to different SRF binding sites. These findings identify myocardin as a master regulator of smooth muscle gene expression and explain how SRF conveys smooth muscle specificity to its target genes. PMID- 12756294 TI - Transcriptome signature of irreversible senescence in human papillomavirus positive cervical cancer cells. AB - A frequent characteristic of human papillomavirus (HPV)-positive cervical cancers is the loss of viral E2 gene expression in HPV-infected cervical epithelial cells as a consequence of viral DNA integration into the cellular genome. The expression of E2 in HPV-positive cancer cells results in the repression of the viral E6/E7 oncogenes, activation of the p53 and pRB pathways, and a G1 cell cycle arrest, followed by induction of cellular senescence. The transcriptional consequences of E2-mediated cell cycle arrest that lead to senescence currently are unknown. Using conditional senescence induction in HeLa cells and microarray analysis, we describe here the expression profile of cells irreversibly committed to senescence. Our results provide insight into the molecular anatomy of senescence pathways and its regulation by HPV on-coproteins. These include the induction of the RAB vesicular transport machinery and a general down-regulation of chromatin regulatory molecules. The repression of tumor-specific G antigens during E2 senescence supports a reversal of the tumorigenic phenotype by E2 and the potential approach of tumor-specific G antigen-specific immunotherapy for cervical cancer. PMID- 12756295 TI - Specific protein methylation defects and gene expression perturbations in coactivator-associated arginine methyltransferase 1-deficient mice. AB - Arginine methylation has been implicated in the regulation of gene expression. The coactivator-associated arginine methyltransferase 1 (CARM1/PRMT4) binds the p160 family of steroid receptor coactivators (SRCs). This association enhances transcriptional activation by nuclear receptors. Here, we show that embryos with a targeted disruption of CARM1 are small in size and die perinatally. The methylation of two known CARM1 substrates, poly(A)-binding protein (PABP1) and the transcriptional cofactor p300, was abolished in knockout embryos and cells. However, CARM1-dependent methylation of histone H3 was not observed. Furthermore, estrogen-responsive gene expression was aberrant in Carm1-/- fibroblasts and embryos, thus emphasizing the role of arginine methylation as a transcription activation tag. These findings provide genetic evidence for the essential role of CARM1 in estrogen-mediated transcriptional activation. PMID- 12756296 TI - Correlated motion and the effect of distal mutations in dihydrofolate reductase. AB - Dihydrofolate reductase (DHFR) catalyzes the reduction of dihydrofolate to tetrahydrofolate. The catalytic rate in this system has been found to be significantly affected by mutations far from the site of chemical activity in the enzyme [Rajagopalan, P. T. R, Lutz, S., and Benkovic, S. J. (2002) Biochemistry 41, 12618-12628]. On the basis of extensive computer simulations for wild-type DHFR from Escherichia coli and four mutants (G121S, G121V, M42F, and M42F/G121S), we show that key parameters for catalysis are changed. The parameters we study are relative populations of different conformations sampled and hydrogen bonds. We find that the mutations result in long-range structural perturbations, rationalizing the effects that the mutations have on the kinetics of the enzyme. Such perturbations also provide a rationalization for the reported nonadditivity effect for double mutations. We finally examine the role a structural perturbation will have on the hydride transfer step. On the basis of our new findings, we discuss the role of coupled motions between distant regions in the enzyme, which previously was reported by Radkiewicz and Brooks. PMID- 12756297 TI - Involvement of multiple signaling pathways in follicular lymphoma transformation: p38-mitogen-activated protein kinase as a target for therapy. AB - Follicular lymphoma (FL) is the most common form of low-grade non-Hodgkin's lymphoma. Transformation to diffuse large B cell lymphoma (DLBCL) is an important cause of mortality. Using cDNA microarray analysis we identified 113 transformation-associated genes whose expression differed consistently between serial clonally related samples of FL and DLBCL occurring within the same individual. Quantitative RT-PCR validated the microarray results and assigned blinded independent group of 20 FLs, 20 DLBCLs, and five transformed lymphoma derived cell lines with 100%, 70%, and 100% accuracy, respectively. Notably, growth factor cytokine receptors and p38beta-mitogen-activated protein kinase (MAPK) were differentially expressed in the DLBCLs. Immunohistochemistry of another blinded set of samples demonstrated expression of phosphorylated p38MAPK in 6/6 DLBCLs and 1/5 FLs, but not in benign germinal centers. SB203580 an inhibitor of p38MAPK specifically induced caspase-3-mediated apoptosis in t(14;18)+/p38MAPK+-transformed FL-derived cell lines. Lymphoma growth was also inhibited in SB203580-treated NOD-SCID mice. Our results implicate p38MAPK dysregulation in FL transformation and suggest that molecular targeting of specific elements within this pathway should be explored for transformed FL therapy. PMID- 12756298 TI - Reversal of hyperglycemia in mice by using human expandable insulin-producing cells differentiated from fetal liver progenitor cells. AB - Beta-cell replacement is considered to be the most promising approach for treatment of type 1 diabetes. Its application on a large scale is hindered by a shortage of cells for transplantation. Activation of insulin expression, storage, and regulated secretion in stem/progenitor cells offers novel ways to overcome this shortage. We explored whether fetal human progenitor liver cells (FH) could be induced to differentiate into insulin-producing cells after expression of the pancreatic duodenal homeobox 1 (Pdx1) gene, which is a key regulator of pancreatic development and insulin expression in beta cells. FH cells possess a considerable replication capacity, and this was further extended by introduction of the gene for the catalytic subunit of human telomerase. Immortalized FH cells expressing Pdx1 activated multiple beta-cell genes, produced and stored considerable amounts of insulin, and released insulin in a regulated manner in response to glucose. When transplanted into hyperglycemic immunodeficient mice, the cells restored and maintained euglycemia for prolonged periods. Quantitation of human C-peptide in the mouse serum confirmed that the glycemia was normalized by the transplanted human cells. This approach offers the potential of a novel source of cells for transplantation into patients with type 1 diabetes. PMID- 12756299 TI - Monocyte chemoattractant protein 1 in obesity and insulin resistance. AB - This study identifies monocyte chemoattractant protein 1 (MCP-1) as an insulin responsive gene. It also shows that insulin induces substantial expression and secretion of MCP-1 both in vitro in insulin-resistant (IR) 3T3-L1 adipocytes and in vivo in IR obese mice (ob/ob). Thus, MCP-1 resembles other previously described genes (e.g., PAI-1 and SREBP-1c) that remain sensitive to insulin in IR states. The hyperinsulinemia that frequently accompanies obesity and insulin resistance may therefore contribute to the altered expression of these and other genes in insulin target tissues. In vivo studies also demonstrate that MCP-1 is overexpressed in obese mice compared with their lean controls, and that white adipose tissue is a major source of MCP-1. The elevated MCP-1 may alter adipocyte function because addition of MCP-1 to differentiated adipocytes in vitro decreases insulin-stimulated glucose uptake and the expression of several adipogenic genes (LpL, adipsin, GLUT-4, aP2, beta3-adrenergic receptor, and peroxisome proliferator-activated receptor gamma). These results suggest that elevated MCP-1 may induce adipocyte dedifferentiation and contribute to pathologies associated with hyperinsulinemia and obesity, including type II diabetes. PMID- 12756300 TI - BRCA2 cooperates with histone acetyltransferases in androgen receptor-mediated transcription. AB - Germ-line mutations of the BRCA2 tumor suppressor gene greatly increase the risk of developing breast and ovarian cancers. Here, we show that wild-type BRCA2, but not a tumor-specific truncated mutant BRCA2, synergizes with the nuclear receptor coactivator p160 GRIP1 to enhance transcriptional activation by androgen receptor (AR). BRCA2 not only associates with AR and GRIP1 but also cooperates with both the histone acetyltransferase P/CAF and BRCA1 to enhance AR- and GRIP1-mediated transactivation. As such, BRCA2 can exert its tumor suppressor function, in part, by modulating androgen signaling, which has been shown to be antiproliferative in a subset of breast cancer cells and particularly implicated in male breast tumors. PMID- 12756301 TI - Seaweed resistance to microbial attack: a targeted chemical defense against marine fungi. AB - Pathogenic microbes can devastate populations of marine plants and animals. Yet, many sessile organisms such as seaweeds and sponges suffer remarkably low levels of microbial infection, despite lacking cell-based immune systems. Antimicrobial defenses of marine organisms are largely uncharacterized, although from a small number of studies it appears that chemical defenses may improve host resistance. In this study, we asked whether the common seaweed Lobophora variegata is chemically defended against potentially deleterious microorganisms. Using bioassay-guided fractionation, we isolated and characterized a 22-membered cyclic lactone, lobophorolide (1), of presumed polyketide origin, with sub-microM activity against pathogenic and saprophytic marine fungi. Deterrent concentrations of 1 were found in 46 of 51 samples collected from 10 locations in the Bahamas over a 4-year period. Lobophorolide (1) is structurally unprecedented, yet parts of the molecule are related to tolytoxin, the scytophycins, and the swinholides, macrolides previously isolated from terrestrial cyanobacteria and from marine sponges and gastropods. Until now, compounds of this structural class have not been associated with marine macrophytes. Our findings suggest that seaweeds use targeted antimicrobial chemical defense strategies and that secondary metabolites important in the ecological interactions between marine macroorganisms and microorganisms could be a promising source of novel bioactive compounds. PMID- 12756302 TI - Hypergeometric generating functions for values of Dirichlet and other L functions. AB - Although there is vast literature on the values of L functions at nonpositive integers, the recent appearance of some of these values as the coefficients of specializations of knot invariants comes as a surprise. Using work of G. E. Andrews [(1981) Adv. Math. 41, 173-185; (1986) q-Series: Their Development and Application in Analysis, Combinatories, Physics, and Computer Algebra, Conference Board of the Mathematical Sciences Regional Conference Series in Mathematics 66 (Am. Math. Soc, Providence, RI); (1975) Problems and Prospects for Basic Hypergeometric Series: The Theory and Application of Special Functions (Academic, New York); and (1992) Illinois J. Math. 36, 251-274], we revisit this old subject and provide uniform and general results giving such generating functions as specializations of basic hypergeometric functions. For example, we obtain such generating functions for all nontrivial Dirichlet L functions. PMID- 12756303 TI - Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation. AB - Multicolor nonlinear microscopy of living tissue using two- and three-photon excited intrinsic fluorescence combined with second harmonic generation by supermolecular structures produces images with the resolution and detail of standard histology without the use of exogenous stains. Imaging of intrinsic indicators within tissue, such as nicotinamide adenine dinucleotide, retinol, indoleamines, and collagen provides crucial information for physiology and pathology. The efficient application of multiphoton microscopy to intrinsic imaging requires knowledge of the nonlinear optical properties of specific cell and tissue components. Here we compile and demonstrate applications involving a range of intrinsic molecules and molecular assemblies that enable direct visualization of tissue morphology, cell metabolism, and disease states such as Alzheimer's disease and cancer. PMID- 12756306 TI - Autogenous bone graft interpositional arthrodesis for the correction of flail toe. A retrospective analysis of 22 procedures. AB - Digital surgery is one of the most common types of surgery performed by foot and ankle surgeons. Flail toe is a complication that may occur after overaggressive resection arthroplasty of the proximal interphalangeal joint of the lesser toes. Correction of flail toe deformity has received little attention and has predominantly involved soft-tissue procedures. The authors' preferred technique for the surgical correction of flail toe is to place a unicortical autogenous bone graft (harvested from the ipsilateral calcaneus) within the revised proximal interphalangeal joint of the lesser toes to create a distraction arthrodesis. This technique allows restoration of digital length, stability, and purchase. A retrospective review of 22 such procedures in 13 patients is presented, along with a literature review of other procedures and a description of the authors' current surgical technique and postoperative management protocol. Overall success using the authors' procedure was 82%. Complications occurred in three patients, with one of the grafts showing complete resorption and two requiring additional surgical intervention owing to nonunion and malunion of toes. PMID- 12756305 TI - 3D electron microscopy reveals the variable deposition and protein dynamics of the peripheral pyruvate dehydrogenase component about the core. AB - Cryo-electron microscopy was exploited to reveal and study the influence of pyruvate dehydrogenase (E1) occupancy on the conformational states of the Saccharomyces cerevisiae pyruvate dehydrogenase complex (PDC). Structures representative of PDC preparations with approximately 40% and full E1 occupancy were determined after the electron microscopy images from each preparation were classified according to their sizes. The reconstructions derived from two size groups showed that the deposition of the E1 molecules associated with the larger complex is, unexpectedly, not icosahedrally arranged, whereas in the smaller complex the E1 molecules have an arrangement and architecture similar to their more ordered deposition in the WT bovine kidney PDC. This study also shows that the linker of dihydrolipamide acetyltransferase (E2) that tethers E1 to the E2 core increases in length from approximately 50 to 75 A, accounting largely for the size difference of the smaller and larger structures, respectively. Extensive E1 occupancy of its 60 E2 binding sites favors the extended conformation of the linker associated with the larger complex and appears to be related to the loss of icosahedral symmetry of the E1 molecules. However, the presence of a significant fraction of larger molecules also in the WT PDC preparation with low E1 occupancy indicates that the conformational variability of the linker contributes to the overall protein dynamics of the PDC and the variable deposition of E1. The flexibility of the complex may enhance the catalytic proficiency of this macromolecular machine by promoting the channeling of the intermediates of catalysis between the active sites. PMID- 12756304 TI - A global view of the selectivity of zinc deprivation and excess on genes expressed in human THP-1 mononuclear cells. AB - Among the micronutrients required by humans, zinc has particularly divergent modes of action. cDNA microarray and quantitative PCR technologies were used to investigate the zinc responsiveness of known genes that influence zinc homeostasis and to identify, through global screening, genes that may relate to phenotypic outcomes of altered dietary zinc intake. Human monocytic/macrophage THP-1 cells were either acutely zinc depleted, using a cell-permeable zinc specific chelator, or were supplemented with zinc to alter intracellular zinc concentrations. Initially, genes associated with zinc homeostasis were evaluated by quantitative PCR to establish ranges for fold changes in transcript abundance that might be expected with global screening. Zinc transporter-1 and zinc transporter-7 expression increased when cellular zinc increased, whereas Zip-2 expression, the most zinc-responsive gene examined, was markedly increased by zinc depletion. Microarrays composed of approximately 22,000 elements were used to identify those genes responsive to either zinc depletion, zinc supplementation, or both conditions. Hierarchal clustering and ANOVA revealed that approximately 5% or 1,045 genes were zinc responsive. Further sorting based on this pattern of the zinc responsiveness of these genes into seven groups revealed that 104 genes were linearly zinc responsive in a positive mode (i.e., increased expression as cellular zinc increases) and 86 genes that were linearly zinc responsive in a negative mode (i.e., decreased expression as cellular zinc increases). Expression of some genes was responsive to only zinc depletion or supplementation. Categorization by function revealed numerous genes needed for host defense were among those identified as zinc responsive, including cytokine receptors and genes associated with amplification of the Th1 immune response. PMID- 12756307 TI - Neurogenic positional pedal neuritis. Common pedal manifestations of spinal stenosis. AB - Neurogenic positional pedal neuritis is a presentation of neuritic symptoms in one or both feet usually affected by body position, specifically, the position of the spine. Its etiology is similar to that of neurogenic-induced claudication caused by spinal stenosis in that the symptoms are caused by compression or irritation of nerves of the lower lumbosacral spine, usually the fifth lumbar and first sacral nerve roots. Burning, stabbing, a cold feeling, aching, numbness, paresthesia, or a weak or tired feeling of the feet (during some part of the disease process) depend on spinal position and may occur during standing, walking, or even lying in bed. Symptoms may be severe and are often eliminated by lumbosacral spine flexion, such as by walking with wheeled support such as a grocery cart or walker; less frequently by negative-heel shoe modification, which can change the position of the lumbosacral spine in stance; or by alteration of sleeping position. This condition, which can include loss of protective sensation, is often misdiagnosed as neuropathy (especially in diabetic patients) or less frequently as biomechanical in origin. In diabetic patients, this condition is frequently the cause of failure of monochromatic infrared energy therapy for diabetic peripheral neuropathy. Treatment is aimed at reducing the spinal nerve or nerve root irritation. Clear definition of the pedal symptoms of spinal nerve compression within a single diagnostic category should facilitate identification and treatment. PMID- 12756308 TI - The reliability of the manual supination resistance test. AB - Several decisions need to be made when prescribing foot orthoses for abnormal foot pronation. One of these decisions is how much force is needed from orthoses to supinate the foot. The supination resistance test has been described as one technique to help determine the amount of force needed. The aim of this project was to determine the reliability of the manual supination resistance test. Four clinicians of differing levels of experience performed the test on 44 subjects (88 feet) on 2 separate days. The test had good reliability overall, with an intertester intraclass correlation coefficient of 0.89. For the two more experienced clinicians, the intratester intraclass correlation coefficients were good (0.82 and 0.78), but for the two inexperienced clinicians they were poor (0.56 and 0.62). The supination resistance test may be clinically useful in the prescription of foot orthoses, but more work is needed to determine its validity and its relationship to clinical outcomes. PMID- 12756309 TI - A modified technique for Morton's neuroma. Decompression with relocation. AB - Interdigital nerve decompression with relocation was performed on 82 feet in 78 patients. The primary indication for surgery was chronic neuritic symptoms that did not resolve with conservative treatment. All but four patients (95%) achieved complete resolution of preoperative symptoms within an average of 7 days following surgery, with full sensation restored at an average of 5 weeks. All of the patients could tolerate a shoe with a wide toe box within 8 days postoperatively. Interdigital nerve decompression with relocation provides for rapid resolution of neuritic symptoms and early return to normal activities. It is also a relatively easy surgical technique. As such, nerve decompression with relocation should be the procedure of choice for the treatment of Morton's neuroma or interdigital nerve compression syndrome. PMID- 12756310 TI - Confirmation of dermatophytes in nail specimens using in-office dermatophyte test medium cultures. Insights from a multispecialty survey. AB - Using data from a multicenter nationwide multispecialty survey, the authors investigated the efficacy of in-office dermatophyte test medium (DTM) and central laboratory cultures used to confirm onychomycosis across samples collected by podiatric, dermatologic, and primary-care physicians. The samples collected by podiatric physicians were both positive or both negative in 43% and 27% of patients, respectively. Samples harvested by dermatologists were both positive in 37% of patients and both negative in 32%, while the samples collected by primary care physicians were both positive in 28% of patients and both negative in 38%. The accuracy of DTM and central laboratory tests is dependent on the proper collection of nail samples, and the accuracy of mycologic test results varied significantly across nail specimens harvested by podiatric, dermatologic, and primary-care physicians. DTM culture was found to be an effective and convenient method of confirming dermatophyte infections in patients with signs of onychomycosis. The data presented here indicate that the special expertise of podiatric physicians in treating foot-related illnesses translates into more accurate mycologic testing. PMID- 12756312 TI - High plantar pressure and callus in diabetic adolescents. Incidence and treatment. AB - This study examined the incidence of high peak plantar pressure and plantar callus in 211 adolescents with diabetes mellitus and 57 nondiabetic controls. The percentage of subjects with these anomalies was the same in both groups. Although diabetic subjects were no more likely than nondiabetic controls to have high peak plantar pressure and callus, these anomalies place individuals with diabetes at greater risk of future foot problems. The effects of orthoses, cushioning, and both in combination were monitored in 17 diabetic subjects with high peak plantar pressure and in 17 diabetic subjects with plantar callus; reductions of up to 63% were achieved. Twelve-month follow-up of diabetic subjects fitted with orthoses showed a significant reduction in peak plantar pressure even when the orthoses were removed. The diabetic subjects who had not received any interventions during the same 12-month period showed no significant change in peak plantar pressure. PMID- 12756311 TI - Reliability of the foot posture index and traditional measures of foot position. AB - Repeatable measures are essential for clinicians and researchers alike. Both need baseline measures that are reliable, as intervention effects cannot be accurately identified without consistent measures. The intrarater and interrater reliability of the new Foot Posture Index and current podiatric measures of foot position were assessed using a same-subject, repeated-measures study design across three age groups. The Foot Posture Index total score showed moderate reliability overall, demonstrating better reliability than most other current measures, although navicular height (normalized for foot length) was the single most reliable measure in adults. None of the tested measures exhibited adequate reliability in young children, and, with less-than-desirable reliability being demonstrated, most measures need to be interpreted accordingly when repeated measures are involved. PMID- 12756313 TI - The fitting of amputated and nonamputated diabetic feet. A French experience at the Villiers-Saint-Denis Hospital. AB - The Villiers-Saint-Denis Hospital in France specializes in the rehabilitation of and fitting of orthoses for lower-limb amputees, who frequently have diabetes mellitus. The percentage of partial-foot amputations has increased relative to the percentage of transtibial or transfemoral amputations. This article describes a complete range of orthoses and prostheses, adapted to each patient, that allow recovery of the standing position, gait ability, and physical activity. PMID- 12756314 TI - Charcot-Marie-Tooth disease and vincristine. AB - This article reports on a case of sensorimotor neuropathy in a 55-year-old man that developed after vincristine therapy. Subsequent biopsy of the sural nerve and electromyographic studies revealed the presence of Charcot-Marie-Tooth disease. Only 17 patients who developed severe neuropathy with very low accumulated doses of vincristine have been described in the literature. Pain and lateral ankle instability were treated with a functional orthosis. Orthopedic treatment and the biomechanical basis of foot and ankle problems in patients with vincristine therapy-induced Charcot-Marie-Tooth disease are discussed. PMID- 12756315 TI - Plantar fasciitis: a degenerative process (fasciosis) without inflammation. AB - The authors review histologic findings from 50 cases of heel spur surgery for chronic plantar fasciitis. Findings include myxoid degeneration with fragmentation and degeneration of the plantar fascia and bone marrow vascular ectasia. Histologic findings are presented to support the thesis that "plantar fasciitis" is a degenerative fasciosis without inflammation, not a fasciitis. These findings suggest that treatment regimens such as serial corticosteroid injections into the plantar fascia should be reevaluated in the absence of inflammation and in light of their potential to induce plantar fascial rupture. PMID- 12756316 TI - Lichen planus. A case study. PMID- 12756317 TI - Human skin equivalent in the treatment of chronic leg ulcers in sickle cell disease patients. PMID- 12756318 TI - Council on Podiatric Medical Education: Eighty-third Annual Report, 2002. PMID- 12756320 TI - The effect of nonsense codons on splicing: a genomic analysis. AB - The phenomenon of nonsense-associated altered splicing raises the possibility that the recognition of in-frame nonsense codons is used generally for exon identification during pre-mRNA splicing. However, nonsense codon frequencies in pseudo exons and in regions flanking 5' splice sites are no greater than that expected by chance, arguing against the widespread use of this strategy as a means of rejecting potential splice sites. PMID- 12756321 TI - On the occurrence of the T-loop RNA folding motif in large RNA molecules. AB - The T-loop RNA folding motif may be considered as a five-nucleotide motif composed of a U-turn flanked by a noncanonical base pair. It was recently proposed that the flanking noncanonical base pair is always a UA trans Watson Crick/Hoogsteen base pair stacked on a Watson-Crick base pair on one side. Here we show that structural analysis of several large RNA molecules, including the recently solved crystal structure of the specificity domain of Bacillus subtilis RNase P, combined with sequence analysis, indicates a broader sequence consensus for the motif. Additionally, we show that the flanking base pair does not necessarily stack on a Watson-Crick base pair and the 3' terminus of the five nucleotide motif is often followed by a sharp turn in the phosphate backbone rather than just a bulged base or bases. PMID- 12756322 TI - Metabolite-binding RNA domains are present in the genes of eukaryotes. AB - Genetic control by metabolite-binding mRNAs is widespread in prokaryotes. These riboswitches are typically located in noncoding regions of mRNA, where they selectively bind their target compound and subsequently modulate gene expression. We have identified mRNA elements in fungi and in plants that match the consensus sequence and structure of thiamine pyrophosphate-binding domains of prokaryotes. In Arabidopsis, the consensus motif resides in the 3'-UTR of a thiamine biosynthetic gene, and the isolated RNA domain binds the corresponding coenzyme in vitro. These results suggest that metabolite-binding mRNAs are possibly involved in eukaryotic gene regulation and that some riboswitches might be representatives of an ancient form of genetic control. PMID- 12756323 TI - Stop codon suppression via inhibition of eRF1 expression. AB - In humans, recognition of a stop codon by protein release factor eRF1 leads to release of the nascent peptide from the ribosome. Although efficient eRF1 activity is usually desirable, numerous pathologies result from eRF1 recognition of premature stop mutations in essential genes. In these cases, decreased eRF1 activity could increase readthrough of the premature stop codon, thereby making full-length protein. To broaden the means available to beneficially decrease eRF1 activity, we have targeted eRF1 mRNA using siRNAs and antisense oligonucleotides. We show that both eRF1-targeted siRNA and antisense oligonucleotides decrease eRF1 mRNA and eRF1 protein concentrations, and increase UAG readthrough in cultured human cells. PMID- 12756324 TI - The interaction of the cap-binding complex (CBC) with eIF4G is dispensable for translation in yeast. AB - In eukaryotes, the m(7)GpppN cap structure is added to all nascent RNA polymerase II transcripts, and serves important functions at multiple steps of RNA metabolism. The predominantly nuclear cap-binding complex (CBC) binds to the cap during RNA synthesis. The predominantly cytoplasmic eukaryotic initiation factor 4F (eIF4F) is thought to replace CBC after export of mature mRNA to the cytoplasm, and mediates the bulk of cellular translation. Yeast as well as mammalian CBC interacts in vitro with eIF4G, a subunit of eIF4F. In this work, we investigate a potential role of this interaction during translation in yeast. We identify a mutation (DR548/9AA) in Tif4631p, one of two isoforms of yeast eIF4G, that abolishes its binding to CBC. Cells expressing this mutant protein as the sole source of eIF4G grow at wild-type rates, and bulk cellular translation, as assessed by metabolic labeling and polysome profile analysis, is unchanged. Importantly, we find that the DR548/9AA mutation neither diminishes nor delays the translation of newly induced reporter mRNA. Finally, microarray analysis reveals marked transcriptome alterations in CBC subunit deletion strains, whereas eIF4G point mutants have essentially a wild-type transcriptome composition. Collectively, these data suggest that in yeast, the phenotypic consequences of CBC deletions are separable from its interaction with eIF4G, and that the CBC eIF4G interaction is dispensable for a potential "pioneering round" of translation in yeast. PMID- 12756325 TI - Structural mimicry in the phage phi21 N peptide-boxB RNA complex. AB - We determined the solution structure of a 22-amino-acid peptide from the amino terminal domain of the bacteriophage phi21 N protein in complex with its cognate 24-mer boxB RNA hairpin using heteronuclear magnetic resonance spectroscopy. The N peptide binds as an alpha-helix and interacts predominately with the major groove side of the 5' half of the boxB RNA stem-loop. This binding interface is defined by surface complementarity of polar and nonpolar interactions, and little sequence-specific recognition. The phi21 boxB loop (CUAACC) has hydrogen bond and backbone torsions typical of the "U-turn" motif, as well as base stacking of the last 4 nt, and a hydrogen bonded C:C pair closing the loop. The exposed face of the phi21 boxB loop, in complex with the N peptide, is strikingly similar to the GNRA tetraloop-like folds of the related lambda and P22 bacteriophage N peptide boxB RNA complexes. The N peptide-boxB complexes of the various phage, while individually distinct, provide similar structural features for interactions with the Escherichia coli host factors to enable antitermination. PMID- 12756327 TI - Substrate specificity and reaction kinetics of an X-motif ribozyme. AB - The X-motif is an in vitro-selected ribozyme that catalyzes RNA cleavage by an internal phosphoester transfer reaction. This ribozyme class is distinguished by the fact that it emerged as the dominant clone among at least 12 different classes of ribozymes when in vitro selection was conducted to favor the isolation of high-speed catalysts. We have examined the structural and kinetic properties of the X-motif in order to provide a framework for its application as an RNA cleaving agent and to explore how this ribozyme catalyzes phosphoester transfer with a predicted rate constant that is similar to those exhibited by the four natural self-cleaving ribozymes. The secondary structure of the X-motif includes four stem elements that form a central unpaired junction. In a bimolecular format, two of these base-paired arms define the substrate specificity of the ribozyme and can be changed to target different RNAs for cleavage. The requirements for nucleotide identity at the cleavage site are GD, where D = G, A, or U and cleavage occurs between the two nucleotides. The ribozyme has an absolute requirement for a divalent cation cofactor and exhibits kinetic behavior that is consistent with the obligate binding of at least two metal ions. PMID- 12756326 TI - An uncapped RNA suggests a model for Caenorhabditis elegans polycistronic pre mRNA processing. AB - Polycistronic pre-mRNAs from Caenohabditis elegans operons are processed by internal cleavage and polyadenylation to create 3' ends of mature mRNAs. This is accompanied by trans-splicing with SL2 approximately 100 nucleotides downstream of the 3' end formation sites to create the 5' ends of downstream mRNAs. SL2 trans-splicing depends on a U-rich element (Ur), located approximately 70 nucleotides upstream of the trans-splice site in the intercistronic region (ICR), as well as a functional 3' end formation signal. Here we report the existence of a novel gene-length RNA, the Ur-RNA, starting just upstream of the Ur element. The expression of Ur-RNA is dependent on 3' end formation as well as on the presence of the Ur element, but does not require a trans-splice site. The Ur-RNA is not capped, and alteration of the location of the Ur element in either the 5' or 3' direction alters the location of the 5' end of the Ur-RNA. We propose that a 5' to 3' exonuclease degrades the precursor RNA following cleavage at the poly(A) site, stopping when it reaches the Ur element, presumably attributable to a bound protein. Part of the function of this protein can be performed by the MS2 coat protein. Recruitment of coat protein to the ICR in the absence of the Ur element results in accumulation of an RNA equivalent to Ur-RNA, and restores trans-splicing. Only SL1, however, is used. Therefore, coat protein is sufficient for blocking the exonuclease and thereby allowing formation of a substrate for trans-splicing, but it lacks the ability to recruit the SL2 snRNP. Our results also demonstrate that MS2 coat protein can be used as an in vivo block to an exonuclease, which should have utility in mRNA stability studies. PMID- 12756328 TI - RNA editing and regulation of Drosophila 4f-rnp expression by sas-10 antisense readthrough mRNA transcripts. AB - We have previously described an example of extensively A-to-G edited cDNA derived from adult heads of the fruitfly Drosophila melanogaster. In that study, the source of the predicted antisense RNA pairing strand for template recognition by dADAR editase was not identified, and the biological significance of the observed hyperediting was not known. Here, we address each of these questions. 4f-rnp and sas-10 are closely adjacent X-linked genes located on opposite DNA strands that produce convergent transcripts. We show that developmentally regulated antisense sas-10 readthrough mRNA arises by activation of an upstream promoter P2 during the late embryo stage of fly development. The sas-10 readthrough transcripts pair with 4f-rnp mRNA to form double-stranded molecules, as indicated by A-to-G editing observed in both RNA strands. It would be predicted that perfect RNA duplexes would be targeted for modification/degradation by enzyme pathways that recognize double-stranded RNAs, leading to decline in 4f-rnp mRNA levels, and this is what we observe. The observation using quantitative RT-PCR that sas-10 readthrough and 4f-rnp transcript levels are inversely related suggests a role for the antisense RNA in posttranscriptional regulation of 4f-rnp gene expression during development. Potential molecular mechanisms that could lead to this result are discussed, one of which is targeted transcript degradation via the RNAi pathway. Insofar as the dADAR editase and RNAi pathways are known to be constitutive in this system, it is likely that control of antisense RNA transcription is the rate-limiting factor. The results provide insight into roles of naturally occurring antisense RNAs in regulation of eukaryotic gene expression. PMID- 12756329 TI - A novel unanticipated type of pseudouridine synthase with homologs in bacteria, archaea, and eukarya. AB - Putative pseudouridine synthase genes are members of a class consisting of four subgroups that possess characteristic amino acid sequence motifs. These genes have been found in all organisms sequenced to date. In Escherichia coli, 10 such genes have been identified, and the 10 synthase gene products have been shown to function in making all of the pseudouridines found in tRNA and ribosomal RNA except for tRNA(Glu) pseudouridine13. In this work, a protein able to make this pseudouridine was purified by standard biochemical procedures. Amino-terminal sequencing of the isolated protein identified the synthase as YgbO. Deletion of the ygbO gene caused the loss of tRNA(Glu) pseudouridine13 and plasmid-borne restoration of the structural gene restored pseudouridine13. Reaction of the overexpressed gene product, renamed TruD, with a tRNA(Glu) transcript made in vitro also yielded only pseudouridine13. A search of the database detected 58 homologs of TruD spanning all three phylogenetic domains, including ancient organisms. Thus, we have identified a new wide-spread class of pseudouridine synthase with no sequence homology to the previously known four subgroups. The only completely conserved sequence motif in all 59 organisms that contained aspartate was GXKD, in motif II. This aspartate was essential for in vitro activity. PMID- 12756330 TI - Effect of transcription on folding of the Tetrahymena ribozyme. AB - Sequential formation of RNA interactions during transcription can bias the folding pathway and ultimately determine the functional state of a transcript. The kinetics of cotranscriptional folding of the Tetrahymena L-21 ribozyme was compared with refolding of full-length transcripts under the same conditions. Sequential folding after transcription by phage T7 or Escherichia coli polymerase is only twice as fast as refolding, and the yield of native RNA is the same. By contrast, a greater fraction of circularly permuted variants folded correctly at early times during transcription than during refolding. Hybridization of complementary oligonucleotides suggests that cotranscriptional folding enables a permuted RNA beginning at G303 to escape non-native interactions in P3 and P9. We propose that base pairing of upstream sequences during transcription elongation favors branched secondary structures that increase the probability of forming the native ribozyme structure. PMID- 12756331 TI - Recognition of the 5' leader of pre-tRNA substrates by the active site of ribonuclease P. AB - The bacterial tRNA processing enzyme ribonuclease P (RNase P) is a ribonucleoprotein composed of a approximately 400 nucleotide RNA and a smaller protein subunit. It has been established that RNase P RNA contacts the mature tRNA portion of pre-tRNA substrates, whereas RNase P protein interacts with the 5' leader sequence. However, specific interactions with substrate nucleotides flanking the cleavage site have not previously been defined. Here we provide evidence for an interaction between a conserved adenosine, A248 in the Escherichia coli ribozyme, and N(-1), the substrate nucleotide immediately 5' of the cleavage site. Specifically, mutations at A248 result in miscleavage of substrates containing a 2' deoxy modification at N(-1). Compensatory mutations at N(-1) restore correct cleavage in both the RNA-alone and holoenzyme reactions, and also rescue defects in binding thermodynamics caused by A248 mutation. Analysis of pre-tRNA leader sequences in Bacteria and Archaea reveals a conserved preference for U at N(-1), suggesting that an interaction between A248 and N(-1) is common among RNase P enzymes. These results provide the first direct evidence for RNase P RNA interactions with the substrate cleavage site, and show that RNA and protein cooperate in leader sequence recognition. PMID- 12756334 TI - Presidential address: medical education: thoughts on the training of physicians and surgeons. PMID- 12756332 TI - In vivo analysis of nucleolar proteins modified by the yeast arginine methyltransferase Hmt1/Rmt1p. AB - In this report, we have investigated the impact of arginine methylation on the Gar1, Nop1, and Nsr1 nucleolar proteins in Saccharomyces cerevisiae. Although previous reports have established that protein arginine methylation is important for nucleocytoplasmic shuttling, they have focused on the examination of heterogeneous nuclear ribonucleoproteins (hnRNPs). We have extended this analysis to several nucleolar proteins that represent a distinct functional class of arginine-methylated proteins. We first developed an in vivo assay to identify proteins methylated by the Hmt1 arginine methyltransferase. This assay is based on the fact that the Hmt1 enzyme utilizes S-Adenosyl-L-methionine as the methyl donor for protein arginine methylation. Following SDS polyacrylamide electrophoresis, 11 distinct proteins were identified as substrates for the Hmt1 methyltransferase. Hmt1p overexpression did not increase the methylation level on these proteins, suggesting they are fully methylated under the conditions examined. Three of the radiolabeled proteins were confirmed to be Gar1p, Nop1p, and Nsr1p. To monitor the cellular localization of these proteins, functional GFP fusion proteins were generated and found to be localized to the nucleolus. This localization was independent of arginine methylation. Furthermore, all three proteins examined did not export to the cytoplasm. In contrast, arginine methylation is required for the export of the nuclear RNA-binding proteins Npl3p, Hrp1p, and Nab2p. The observation that three nucleolar proteins are modified by Hmt1p but are not exported from the nucleolus implies an alternate role for arginine methylation. PMID- 12756333 TI - Transfer RNA modifications that alter +1 frameshifting in general fail to affect 1 frameshifting. AB - Using mutants (tgt, mnmA(asuE, trmU), mnmE(trmE), miaA, miaB, miaE, truA(hisT), truB) of either Escherichia coli or Salmonella enterica serovar Typhimurium and the trm5 mutant of Saccharomyces cerevisiae, we have analyzed the influence by the modified nucleosides Q34, mnm(5)s(2)U34, ms(2)io(6)A37, Psi39, Psi55, m(1)G37, and yW37 on -1 frameshifts errors at various heptameric sequences, at which at least one codon is decoded by tRNAs having these modified nucleosides. The frequency of -1 frameshifting was the same in congenic strains only differing in the allelic state of the various tRNA modification genes. In fact, in one case (deficiency of mnm(5)s(2)U34), we observed a reduced ability of the undermodified tRNA to make a -1 frameshift error. These results are in sharp contrast to earlier observations that tRNA modification prevents +1 frameshifting suggesting that the mechanisms by which -1 and +1 frameshift errors occur are different. Possible mechanisms explaining these results are discussed. PMID- 12756336 TI - Transrenal fixation of endovascular stent-grafts for infrarenal aortic aneurysm repair: mid-term results. AB - PURPOSE: We evaluated mid-term results of a single-center consecutive series of endovascular stent-grafts implanted for aortic aneurysm repair with transrenal fixation, to determine clinical outcome, aneurysm anatomy, renal artery patency, and renal complications. METHODS: Modular stent-grafts were placed with transrenal fixation in 37 patients between November 1998 and July 2000. Follow-up evaluation included clinical examination, laboratory evaluation of serum creatine concentration, computed tomographic angiography, and renal duplex scanning. RESULTS: Thirty-seven patients underwent transrenal fixation of aortic stent grafts as part of a Phase II US Food and Drug Administration study. Two patients subsequently underwent follow-up at institutions closer to their homes, and thus provided clinical information but no long-term renal or aneurysm morphologic data. There were no perioperative deaths. Five patients died during follow-up, at a mean of 9 months, because of myocardial infarction in 4 patients and respiratory failure in 1 patient. Thirty patients, ages 75 +/- 8 years, have been followed up for 28.5 +/- 7.2 months. Aneurysm diameter at follow-up was 5.0 +/- 0.8 cm, compared with 5.7 +/- 0.8 cm preoperatively. In 5 patients, endoleak developed during follow-up: 1 type I leak was treated with an aortic cuff, with temporary stabilization of the aneurysm and correction of the endoleak; 2 type II endoleaks were treated with translumbar coil embolization, and 1 resolved spontaneously; and 1 type III endoleak was treated with a combination of coil embolization and stent-graft extension to cover a graft defect. Preoperatively, serum creatinine concentration was normal in 23 patients, but increased persistently in 2 patients and was abnormal in 7 patients. Postoperatively, creatine concentration increased in 4 patients to greater than 20% of baseline level. Seventeen patients had no evidence of renal artery stenosis, compared with 13 patients with renal artery stenosis. Of 41 normal renal arteries, 90% remained unchanged, 1 became occluded, 3 demonstrated 60% stenosis. Nephrectomy was necessary in 1 patient because of cancer. Of 19 abnormal renal arteries, progression of disease was noted in 3 arteries. CONCLUSIONS: Transrenal fixation of aortic stent-grafts can be performed with acceptable mid-term outcome with respect to mortality, need for follow-up intervention, and aneurysm exclusion with protection from rupture. Postprocedural stenosis can develop in both normal and abnormal renal arteries. Rate of progression of disease was greater in patients with preprocedural renal dysfunction compared with patients with normal renal arteries. This is merely an observation, and may not be related to transrenal fixation. Long-term follow up is needed. PMID- 12756335 TI - Nature, frequency, and predictors of secondary procedures after endovascular repair of abdominal aortic aneurysm. AB - OBJECTIVES: Endovascular stent grafting offers a potentially less invasive option for treatment of abdominal aortic aneurysm. Clinical benefit has been demonstrated with respect to early parameters such as blood transfusion, return of gastrointestinal function, and length of hospital stay. Endovascular repair, however, has been criticized on the basis of inferior long-term outcome. Secondary procedures may be necessary to address durability issues such as migration, high-pressure endoleak, graft limb thrombosis, and degeneration of the stent-fabric structure itself, issues that may compromise the primary goal of aneurysm repair, protection from rupture. METHODS: Between 1996 and 2002, 703 patients underwent endovascular treatment of infrarenal abdominal aortic aneurysm at The Cleveland Clinic Foundation. During this time, five devices were used: Ancure, AneuRx, Excluder, Talent, and Zenith. Outcome was assessed with physical examination, lower extremity arterial studies, plain abdominal radiography, and computed tomography at discharge, at 1, 6, and 12 months postoperatively, and annually thereafter. Secondary procedures were defined as any procedure, exclusive of diagnostic angiography, performed after stent graft implantation, directed at treatment of aneurysm-related events. Multivariable statistical techniques for censored data (Cox proportional hazards modeling) were used to determine baseline parameters associated with need for secondary procedures over follow-up, with calculation of hazards ratio (HR) and 95% confidence interval (CI). RESULTS: Patient follow-up averaged 12.2 +/- 11.7 months. Patient survival was 90% +/- 1.4% at 1 year, 78% +/- 2.6% at 2 years, and 70% +/- 3.8% at 3 years. Aneurysm rupture occurred in 3 patients (0.4%), accounting for rupture risk of 1.4% over the first 2 years of follow-up (Kaplan-Meier method). Overall, 128 secondary procedures were required in 104 patients (15%), with a cumulative risk of 12% +/- 1.5% at 1 year, 24% +/- 2.8% at 2 years, and 35% +/- 4.4% at 3 years after stent graft implantation. Among the secondary procedures, new stent grafts and extensions were placed in 34 patients (27%), embolization of endoleak was performed in 33 patients (26%), and open surgical conversion was undertaken in 11 patients (9%). Periprocedural mortality of secondary procedures was 8% overall, but was 18% for patients undergoing open surgical conversion. Multivariable modeling identified the date the procedure was performed (HR, 1.53 per 3-month period of study; CI, 1.22-1.92; P <.001) and aneurysm size (HR, 1.35 per centimeter of minor axis; CI, 1.13-1.60; P <.001) as independent predictors of need for secondary procedures. CONCLUSIONS: Current endovascular devices are associated with a relatively high rate of complications over mid-term follow-up, culminating in frequent need for secondary remedial procedures. With strict follow-up imaging compliance, however, risk for rupture and aneurysm-related death remain exceedingly low. Newer technology may achieve improved durability and a lower requirement for secondary procedures, while maintaining the minimally invasive nature of presently available devices. PMID- 12756338 TI - Relationship of residual intraluminal to intrathrombotic pressure in a closed aneurysmal sac. AB - PURPOSE: This study was undertaken to determine the relationship of residual intraluminal aneurysmal sac pressure (ILASP) to intrathrombic aneurysm sac pressure (ITASP) and to define the relationship between abdominal aorta aneurysm (AAA) size (anteroposterior or transverse diameter), volume of intraluminal thrombus, and residual ITASP. METHODS: We measured ILASP and ITASP after proximal aortic neck and distal iliac clamping by placing angiocatheters into the lumen and thrombus of an excluded aneurysm sac in 41 consecutive patients. Simultaneously, mean blood pressure was recorded and aneurysm sac pressure ratio was calculated. Changes in ILASP and ITASP after clamping of the inferior mesenteric artery were recorded. In addition, correlation between AAA size, volume of intraluminal thrombus in AAA, and residual ITASP was determined. RESULTS: Mean ILASP/blood pressure ratio was 0.40 (SD, 0.20). Mean ITASP/blood pressure ratio was 0.37 (SD, 0.23). There was a significant positive correlation of 0.47 between ITASP and ILASP (P =.002). Clamping of the inferior mesenteric artery resulted in markedly decreased ITASP in 2 patients (n = 40) and ILASP in 4 patients (n = 41). Each centimeter increase in AAA size resulted in a 47 mL increase in thrombus volume. CONCLUSION: Increased ILASP results in corresponding increase in ITASP, and increased AAA size is associated with increased thrombus volume. However, neither thrombus volume nor AAA size has any relationship to ITASP. PMID- 12756337 TI - Distal internal iliac artery embolization: a procedure to avoid. AB - OBJECTIVES: Internal iliac artery (IIA) coil embolization as an adjunct to endovascular stent grafting (ESG) is common practice for treating abdominal aortic aneurysm (AAA) in patients with a substantially enlarged common iliac artery requiring extension of the stent-graft limb into the external iliac artery. The literature describing pelvic ischemia in association with IIA coil embolization contains conflicting reports of symptom severity. We studied IIA occlusion outcome as a function of coil placement in the IIA. METHODS: From August 1997 to March 2002, 20 patients with AAA underwent ESG with unilateral IIA coil embolization. Coils were placed proximal to the first branch of the IIA in 8 patients and distal to the first branch in 12 patients. Symptoms of pelvic ischemia and mid-term outcome were studied. RESULTS: Patients included 18 men and 2 women with mean age of 70(1/2) years (range, 53-86 years). Mean diameter of AAA was 54.4 mm (range, 38-80 mm), and of common iliac artery was 24.2 mm (range, 15 48 mm). Ten patients (50%) had new onset of symptoms of pelvic ischemia after endograft procedures: 1 of 8 patients (13%) with proximal IIA embolization had buttock claudication, and 9 of 12 patients (75%) with distal IIA embolization had pelvic ischemic symptoms, including buttock claudication in 8 and impotence in 1 (P =.02, Fisher exact test). No colonic ischemia occurred in this series. At 12 month follow-up, 4 patients with distal IIA embolization were symptom-free. At further follow-up to 24 months, 4 patients remained significantly limited with symptoms of claudication. CONCLUSIONS: A significantly higher incidence of symptoms of pelvic ischemia occurred with more distal placement of coils for IIA embolization. Failure to control for extent of coil placement may account for the apparently conflicting results in published studies. IIA coil embolization should be performed as proximal as possible to prevent interference with pelvic collateral circulation. PMID- 12756339 TI - Fate of excluded popliteal artery aneurysms. AB - OBJECTIVE: Popliteal artery aneurysms (PAA) are frequently treated with ligation and exclusion bypass grafting. It is assumed that these aneurysms will shrink and remain asymptomatic. This may not always be true. We sought to elucidate the fate of excluded PAA over time. METHODS: Data for all PAAs treated with ligation and exclusion bypass grafting between 1986 and 1999 were retrospectively reviewed. Computed tomography (CT) scans and duplex ultrasound scans provided aneurysm patency data and maximal transverse diameter measurements of the popliteal artery during late postoperative follow-up. This information was compared with that from similar preoperative studies. RESULTS: Forty-one patients (39 men, 2 women) underwent 57 ligation and exclusion bypass grafting procedures. Both preoperative and late postoperative (mean, 4.0 years; range, 0.43-13.5 years) CT scans or duplex ultrasound scans were available for review of 25 PAAs in 18 patients (ages 42-80 years; mean, 63 years). Preoperative PAA size ranged from 14 to 45 mm (mean, 28.7 mm). In late follow-up, 12 (48%) PAA had decreased in size (mean, 7.3 mm), 5 (20%) remained unchanged, and 8 (32%) increased in mean transverse diameter (mean, 5.9 mm). One large aneurysm increased by 50%. Contrast material enhancement was identified in the excluded sac in 11 aneurysms. CONCLUSIONS: PAA treated with ligation and exclusion bypass grafting often expand and can become symptomatic. This may be analogous to type II endoleak or endotension noted after aortic endovascular repair. We recommend PAA excision or endoaneurysmorrhaphy when feasible. PMID- 12756341 TI - Hospital volume-related differences in aorto-bifemoral bypass operative mortality in the United States. AB - PURPOSE: Aorto-bifemoral bypass (AFB) is commonly performed in US hospitals. Durable long-term outcome is achieved after AFB performed to treat aortoiliac occlusive disease. However, short-term outcome for complex surgical procedures is not uniform across medical centers. The objective of the current study was to define the relationship of hospital volume to operative mortality after AFB. METHODS: The study included 3073 patients with a primary procedure code for AFB and a diagnostic code for peripheral vascular occlusive disease who received treatment during 1997 at 483 hospitals in the Nationwide Inpatient Sample (NIS). The NIS represents a 20% stratified random sample representative of all US hospitals. Unadjusted and case mix-adjusted analyses were performed. RESULTS: Overall AFB-related mortality was 3.3%. Hospitals that performed more than 25 AFB per year (33% of patients at 37 hospitals in the NIS) had a lower crude mortality rate (3.7% vs 2.2%) compared with hospitals that performed fewer AFB. In a multivariate analysis adjusting for case mix, AFB at a high- volume hospital was associated with 42% decreased risk for in-hospital mortality (odds ratio [OR], 0.58; 95% confidence interval [CI], 0.34-0.97; P =.04) compared with low-volume hospitals. Two other important risk factors associated with increased mortality in the multivariate analysis included age more than 65 years (OR, 3.3; 95% CI, 2.0-5.4) and history of chronic pulmonary disease (OR, 1.9; 95% CI, 1.2-2.9). CONCLUSIONS: AFB operative mortality was significantly lower at high-volume hospitals in this nationally representative database. The effect of hospital volume of AFB procedures on outcome should be of importance to patients, providers, and health policy makers. PMID- 12756340 TI - Reliability of common femoral artery hemodynamics in assessing the severity of aortoiliac inflow disease. AB - OBJECTIVES: We investigated the utility of color duplex ultrasound (CDU)-derived common femoral artery (CFA) hemodynamics for detecting significant aortoiliac occlusive disease and predicting its severity. METHODS: From January 1997 to June 2001, 132 consecutive patients with lower extremity arterial insufficiency underwent both femoropopliteal CDU scanning and aortography with runoff studies. CDU-derived CFA waveform contour (monophasic, biphasic, or triphasic), peak systolic velocity (PSV), and acceleration time were recorded for each patient. Severity of aortoiliac occlusive disease was classified by arteriography into three distinct groups: normal or minimal disease (<50%, group 1), significant focal or diffuse stenoses (>/=50%, group 2), or total occlusion (group 3). Using probability and receiver operating characteristic curve analysis, waveform contour and PSV were compared alone and in combination with the arteriographic groups to identify waveform contours and threshold PSV, which may accurately differentiate the three categories of aortoiliac occlusive disease. RESULTS: Of 214 limbs available for study, 112 composed group 1, 70 composed group 2, and 32 composed group 3. Concomitant femoropopliteal disease was present in 47% of limbs in group 1, 53% of limbs in group 2, and 34% of limbs in group III. An abnormal CFA waveform contour (monophasic or biphasic) differentiated group 1 from groups 2 and 3, with 95% sensitivity, 89% specificity, 89% positive predictive value (PPV), 95% negative predictive value (NPV), and 92% accuracy. Mean PSV and acceleration time for monophasic and biphasic waveforms were 39 cm/sec +/- 19, 178 msec +/- 36 vs 95 cm/sec +/- 67, 97 msec +/- 31 respectively (P <.05). In differentiating between groups 2 and 3, the specificity, PPV, and accuracy for CFA PSV of 70 years) in the common femoral vein, superficial femoral vein, popliteal vein, and posterior tibial vein. The highest augmentation response was found in Asian subjects, in the common and superficial femoral veins and the sapheno-femoral and sapheno-popliteal junctions; and the smallest augmentation response was found in African American subjects, in these same veins and junctions. Differences in vein diameters may explain these findings, ie, smaller diameters in Asians and larger diameters in African Americans. Most important, compared with normal values, augmentation response was decreased in legs with venous obstructive disease only when trophic changes were present. CONCLUSION: Like quantification of reflux, quantitative evaluation of the augmentation response may help in diagnosis of venous obstructive disease when trophic changes are present. PMID- 12756355 TI - A nonintrinsic regional basis for increased infrarenal aortic MMP-9 expression and activity. AB - OBJECTIVE: This investigation was undertaken to determine whether intrinsic or regional factors at different anatomic sites of the aorta affect expression and activity of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs). METHODS: Aortas from Sprague-Dawley rats (n = 22) were divided into arch, descending thoracic, and infrarenal abdominal segments. Specimens were stimulated with interleukin-1beta (IL-1beta) (2 ng/mL) for 72 hours. In separate experiments, syngeneic aortic segments were transplanted from the thoracic or abdominal aortas of donor rats into the infrarenal aortic position of recipient rats (n = 12 each). At 4 weeks, aortas from rats who had received transplants were harvested, sectioned into arch, thoracic, and transplanted thoracic or transplanted abdominal segments, and stimulated with IL 1beta. Reverse transcriptase polymerase chain reaction, zymography, and reverse zymography were performed to assess MMP-9, MMP-2, and TIMP-1 in all aortic segments. Differences were assessed with analysis of variance (ANOVA) and post hoc Tukey test. RESULTS: In control rats, abdominal segments had significantly higher MMP-9 expression compared with arch and thoracic segments (P <.002). Total MMP-9 activity was also higher in abdominal segments (P <.02). In rats who received transplants, transplanted thoracic (P <.004) and transplanted abdominal (P <.05) segments demonstrated upregulation of MMP-9 expression, compared with control arch and thoracic segments. Zymography documented increased total MMP-9 activity in transplanted thoracic (P <.03) and transplanted abdominal (P <.04) segments versus arch and thoracic segments. No significant difference in MMP-9 expression was found between control abdominal, transplanted thoracic, or transplanted abdominal segments. No significant differences in MMP-2 or TIMP-1 expression or activity were demonstrated in either control or transplanted segments. CONCLUSIONS: These data demonstrate that variations in aortic MMP-9 expression and activity result from regional factors affecting the aorta rather than intrinsic aortic wall differences. Increases in abdominal aortic MMP-9 may contribute to the predilection for aneurysm to develop in the infrarenal aorta. PMID- 12756356 TI - Wall shear stress and strain modulate experimental aneurysm cellularity. AB - OBJECTIVE: Clinical evidence indicates that hemodynamic conditions influence abdominal aortic aneurysm (AAA) disease. We modified blood flow to evaluate the effects of wall shear stress (WSS) and relative wall strain (RWS) on aneurysm structure and cellularity. METHODS: Rodent AAAs were created with porcine pancreatic elastase infusion. In group 1 AAA WSS was increased with left femoral arteriovenous fistula creation, whereas in group 2 AAA WSS was decreased with left iliac artery ligation. Aortic flow, wall motion, and blood pressure were recorded in both groups. AAA diameter, endothelial and smooth muscle cellularity (CD31 and alpha-smooth muscle actin immunostaining), markers for cell proliferation (5-bromodeoxyuridine), endothelial and smooth muscle cell growth factor production (vascular endothelial growth factor-D and platelet-derived growth factor-beta, respectively), and apoptosis (deoxyuridine triphosphate nick end-labeled [TUNEL] stain) were compared between groups when the animals were killed. RESULTS: Arteriovenous fistula creation increased WSS (high-flow AAA) by 300% and RWS by 150%. Iliac ligation reduced WSS (low-flow AAA) by 60%. Neither procedure significantly altered systolic, diastolic, or mean aortic pressure. When the animals were killed 7 days after elastase infusion, low-flow AAAs were significantly larger than high-flow AAAs. High-flow AAAs also contained more endothelial cells and smooth muscle cells, and evidence of increased growth factor production, cell proliferation, and decreased apoptosis. No difference in type or severity of AAA inflammatory cell infiltrate was noted between groups. CONCLUSIONS: High flow conditions stimulate endothelial cell and smooth muscle cell proliferation in experimental aneurysms. Enhanced cellularity may stabilize aortic integrity, limiting aneurysm growth. Increased lower extremity activity may prevent or retard AAA disease through salutary effects on aortic remodeling mediated by endothelial cells and smooth muscle cells. PMID- 12756357 TI - R136K fibroblast growth factor-1 mutant induces heparin-independent migration of endothelial cells through fibrin glue. AB - OBJECTIVES: R136K is a mutation of fibroblast growth factor-1 (FGF-1) in which arginine replaces lysine at the primary thrombin cleavage site. This may be important in vivo in inducing endothelial cell (EC) migration and coverage of arterial injury sites by allowing R136K to be used in a fibrin glue delivery system, without thrombin-induced degradation, in the absence of heparin. The objectives of this study were to determine whether R136K, with and without heparin, can induce migration of EC and smooth muscle cells (SMC) through fibrin glue, and to compare these results with those of wild-type FGF-1; and to determine the resistance of R136K to thrombin-induced degradation versus FGF-1. METHODS: The dose-response migration through fibrin glue induced by wild-type FGF 1 and the R136K mutant in the presence and absence of heparin was tested with EC and SMC. Migration was tested with 50, 100, and 200 ng/mL of both FGF-1 and R136K, either with or without 5 U/mL of heparin. Migration of EC was also assessed after growth inhibition with mitomycin C. A novel modified Boyden chamber-type migration assay using fibrin glue on the upper surface of the chamber filter was used to test migration. The fluorescent marker calcein was used to identify those cells that had migrated through the fibrin glue and were embedded in the filter. Molecular degradation by thrombin was assessed with sodium dodecylsulfate polyacrylamide gel electrophoresis. RESULTS: For EC, R136K in the absence of heparin induced significantly more migration than did FGF-1 at 50 (P <.002), 100 (P <.0001), and 200 (P <.0001) ng/mL. In the presence of heparin, a chemotactic response of EC to cytokine was seen at all doses, with no significant difference between FGF-1 and R136K. A dose-dependent difference was noted in this group between the 100 and 200 ng/mL concentrations of cytokine (for FGF-1, P <.0001; for R136K, P <.0001). SMC showed no difference in migration with FGF-1, R136K, or negative control at any dose in the presence or absence of heparin. Gel electrophoresis demonstrated that R136K was more resistant to thrombin degradation than was FGF-1. CONCLUSION: Site-directed mutagenesis of FGF 1 to R136K enables induction of heparin-independent migration of EC through fibrin glue at an optimal concentration of 100 ng/mL. Neither FGF-1 nor R136K elicits SMC migration through fibrin glue. The ability of R136K to induce EC migration through fibrin glue in the absence of heparin may prove useful in vivo by inducing EC migration and coverage of arterial injury sites, thus potentially reducing thrombogenicity and intimal hyperplasia. PMID- 12756359 TI - Endovascular exclusion of iliac artery to iliac vein fistula after lumbar disk surgery. AB - Iliac arteriovenous (AV) fistula is rare after lumbar disk surgery. Traditionally, open repair through the arterial lumen was performed. We report endovascular exclusion of an iliac AV fistula in a 41-year-old woman 8 years after lumbar diskectomy. An angiogram showed an AV fistula connecting the right common iliac artery and vein. This was repaired with placement of two covered wall stents in the right common artery and external iliac artery, and embolization of the right internal iliac artery. Contrast medium-enhanced computed tomography scan at 5 months confirmed elimination of the AV fistula and right iliac artery patency. This technique should be considered in management of iliac AV fistulas. PMID- 12756358 TI - Neutrophil survival on biomaterials is determined by surface topography. AB - PURPOSE: Cardiovascular device-centered infections are a major cause of hospital morbidity, mortality, and expense. Caused by opportunistic bacteria, this phenomenon is thought to arise because of a defect in neutrophil bacterial killing. We have shown that neutrophils that adhere to polystyrene remain viable, whereas neutrophils that adhere to the vascular biomaterials expanded polytetrafluoroethylene (ePTFE) and Dacron undergo a rapid nonapoptotic death. This study was designed to test the hypothesis that surface topography is a determinant of the nonapoptotic death response of neutrophils to biomaterials. METHODS: We took advantage of the ease with which a polystyrene surface can be manipulated to examine the effect of surface topography on neutrophil viability. Neutrophils were exposed to smooth or roughened polystyrene surfaces both in vivo and in vitro. Changes in cell membrane permeability and production of reactive oxygen species by individual cells were monitored with fluorescent dyes. RESULTS: Host cells and isolated human neutrophils died rapidly after adhesion to roughened polystyrene. Neutrophils adherent to roughened surfaces produced more reactive oxygen intermediates than those adherent to smooth surfaces and were first to die. The cell death response precipitated by expanded polytetrafluoroethylene, Dacron, or the roughened surfaces was significantly reduced with treatment of the neutrophils with catalase, diphenylene iodonium, or the src kinase inhibitor PP2 before adhesion. CONCLUSIONS: Neutrophil adhesion to roughened materials triggers rapid production of reactive oxygen species and precipitates a nonapoptotic cell death. Understanding the material properties that trigger these responses is essential to development of the next generation of implantable biomaterials. PMID- 12756360 TI - Multiple isolated aneurysms in a case of "burned out" Takayasu aortitis. AB - Takayasu aortitis (TA) is a chronic inflammatory disease predominantly seen in young Asian women. The disease is idiopathic and largely affects the aorta and its major branches. The basic pathologic changes in TA are fibrosis and subsequent occlusion of the large arteries. TA is classically termed "pulseless" disease, with manifestations during the occlusive stage including limb ischemia, renovascular hypertension, and heart failure. Arterial dilation and aneurysm are largely unappreciated manifestations of TA, but they occur in as many as 32% of affected patients. We report chronic "burned out" TA in a 23-year-old Hispanic woman with isolated aneurysms of the descending thoracic aorta, abdominal aorta, and common iliac arteries, without occlusive disease. PMID- 12756361 TI - Abdominal aortic aneurysm associated with crossed renal ectopia without fusion: case report and literature review. AB - Genitourinary anomalies can present a formidable challenge to the vascular surgeon at abdominal aortic reconstruction. We saw a case of crossed renal ectopia without fusion, a rare anomaly, associated with abdominal aortic aneurysm. Because of risk for injury to the kidney during surgery, preoperative evaluation of this anomaly must include computed tomography, angiography, and intravenous pyelography. Preoperative placement of a ureteral catheter may prevent injury to the anomalous ureter. Renal failure of the ectopic kidney during aortic reconstruction can be a serious problem. We used in situ hypothermic perfusion with cold (4 degrees C) Ringer solution for renal protection, and reimplanted the aberrant renal artery. The postoperative course was good, without major complications. The procedure for renal preservation must be selected on the basis of anatomic findings. We review the literature and present the first case of crossed renal ectopia. PMID- 12756362 TI - Acute limb ischemia secondary to myositis-induced compartment syndrome in a patient with human immunodeficiency virus infection. AB - Myositis, while uncommon, develops more frequently in patients with human immunodeficiency virus infection. We report a case of acute lower leg ischemia caused by myositis in such a patient. Urgent four-compartment fasciotomy of the lower leg was performed, which decompressed the compartmental hypertension and reversed the arterial ischemia. This case underscores the importance of recognizing compartment syndrome as a cause of acute limb ischemia. PMID- 12756363 TI - Guidelines for the treatment of abdominal aortic aneurysms. Report of a subcommittee of the Joint Council of the American Association for Vascular Surgery and Society for Vascular Surgery. AB - Decision-making in regard to elective repair of abdominal aortic aneurysms (AAA) requires careful assessment of factors that influence rupture risk, operative mortality, and life expectancy. Individualized consideration of these factors in each patient is essential, and the role of patient preference is of increasing importance. It is not possible or appropriate to recommend a single threshold diameter for intervention which can be generalized to all patients. Based upon the best available current evidence, 5.5 cm is the best threshold for repair in an "average" patient. However, subsets of younger, good-risk patients or aneurysms at higher rupture risk may be identified in whom repair at smaller sizes is justified. Conversely, delay in repair until larger diameter may be best for older, higher-risk patients, especially if endovascular repair is not possible. Intervention at diameter <5.5 cm appears indicated in women with AAA. If a patient has suitable anatomy, endovascular repair may be considered, and it is most advantageous for older, higher-risk patients or patients with a hostile abdomen or other technical factors that may complicate standard open repair. With endovascular repair, perioperative morbidity and recovery time are clearly reduced; however, there is a higher reintervention rate, increased surveillance burden, and a small but ongoing risk of AAA rupture. There is no justification at present for different indications for endovascular repair, such as earlier treatment of smaller AAA. Until long-term outcome of endoluminal repair is better defined and results of randomized trials available, the choice between endovascular and open repair will continue to rely heavily on patient preference. PMID- 12756364 TI - Computational modeling of arterial biomechanics: insights into pathogenesis and treatment of vascular disease. AB - We review how advances in computational techniques are improving our understanding of the biomechanical behavior of the healthy and diseased cardiovascular system. Numerical modeling of biomechanics is being used in a wide variety of ways, including assessment of effects of mural and hemodynamically induced stresses on atherogenesis, development of risk measures for aneurysm rupture, improvement in interpretation of medical images, and quantification of oxygen transport in diseased and healthy arteries. Although not amenable to routine clinical use, numerical modeling of cardiovascular biomechanics is a powerful research tool. PMID- 12756365 TI - Suprainguinal deep venous thrombosis and pulmonary embolism--common but often silent. PMID- 12756366 TI - Superior mesenteric artery aneurysm reconstruction. PMID- 12756367 TI - Advanced age, dementia, and an abdominal aneurysm: intervene? PMID- 12756369 TI - Regarding "Treatment of type II endoleaks after endovascular repair of abdominal aortic aneurysm". PMID- 12756368 TI - Regarding "Hemodynamic benefits of regional anesthesia for carotid endarterectomy". PMID- 12756371 TI - The importance of philanthropy in sustaining pediatric academic departments. PMID- 12756372 TI - The challenge pays off: early enhanced nutritional intake for VLBW small-for gestation neonates improves long-term outcome. PMID- 12756373 TI - Exhaled nitric oxide: a useful aide in pediatric asthma management? PMID- 12756374 TI - Catch-up growth of head circumference of very low birth weight, small for gestational age preterm infants and mental development to adulthood. AB - OBJECTIVE: To examine the influence of postnatal energy quotient (EQ, energy intake/kg body weight per day) on head circumference (HC) growth and mental development of very low birth weight (VLBW), small for gestational age (SGA, <10th percentile) preterm infants. STUDY DESIGN: SGA VLBW preterm infants (n = 46) with primarily symmetric intrauterine growth restriction were compared with 62 appropriate for gestational age (AGA) VLBW preterm infants and 73 term infants from the Bonn Longitudinal study. RESULTS: Twenty-seven of 46 (59%) of the SGA preterm infants showed complete HC catch-up growth by the age of 12 months, but mostly before 6 months after term (HC catch-up group). These infants had significantly higher mean EQs from day 2 to 10 than the group of 19 infants without HC catch-up (EQ, 95 vs 78). Mean EQs correlated significantly with developmental and intelligence quotients (DQ/IQ) from 18 months to 6 years. As adults, the HC of the HC catch-up group was not significantly different from that of the AGA preterm infants, the term infants, and their parents. The group without HC catch-up had smaller HC as adults. CONCLUSIONS: Our data suggest that early postnatal high-energy nutrient intake for SGA preterm infants is needed to promote HC catch-up growth and to prevent negative consequences of undernutrition. PMID- 12756376 TI - Long-term follow-up of home mechanical ventilation in young children with spinal cord injury and neuromuscular conditions. AB - OBJECTIVES: To provide outcomes of two decades of experience in home ventilation of children with spinal cord injury and neuromuscular conditions. STUDY DESIGN: Data were collected through chart review and interviews on 39 children who had become ventilator-dependent before their 6th birthday; 23 children had neuromuscular diseases and 16 had spinal cord injuries. RESULTS: Patients required an average of 0.7 rehospitalizations per year. There were 8 deaths. Survival rates were 97% at 1 year, 97% at 3 years, 84% at 5 years, and 71% at 10 years. Thirty children attended school, 13 were in regular school (1 at university level), 5 were home-schooled, 5 were in special education schools, and 5 were in regular school with some special education classes. One graduated high school, and another graduated university and received a graduate degree. Three children had progressive weakness. Two gained significant muscle strength. CONCLUSIONS: Our experience showed that these patients can be discharged to home with low morbidity and mortality rates and successful reintegration into the community. PMID- 12756377 TI - Initiation of home mechanical ventilation in children with neuromuscular diseases. AB - OBJECTIVE: To determine how often home mechanical ventilation (HMV) is instituted electively in children with respiratory failure from neuromuscular diseases and whether there were opportunities to discuss therapeutic options with patients/families before respiratory failure. METHODS: Patients with neuromuscular disease (n = 73) requiring HMV (age, 2 months to 24 years) were studied. Whether HMV was initiated nonelectively because of acute respiratory failure or electively before acute respiratory failure, and opportunities for health care providers to discuss therapeutic options with patients/families before acute respiratory failure (hospitalization with pneumonia, clinic visits for preoperative evaluation, pulmonary function testing [PFT] and/or polysomnography [PSG]) were recorded. RESULTS: HMV was initiated electively in 21% of patients with neuromuscular disease; 69% of the nonelective HMV group had HMV initiated after respiratory failure caused by pneumonia. In the nonelective group, opportunities for discussion of therapeutic options with the patients and families could have occurred before respiratory failure during 111 hospitalizations for pneumonia, 13 preoperative evaluations, 43 abnormal PFTs, and 24 abnormal PSGs. CONCLUSIONS: Most patients with neuromuscular disease had HMV initiated nonelectively after acute respiratory failure caused by pneumonia. Opportunities for discussing the therapeutic options with patients and families before respiratory failure were missed or ineffective. PMID- 12756375 TI - Relations between exhaled nitric oxide and measures of disease activity among children with mild-to-moderate asthma. AB - OBJECTIVE: Exhaled nitric oxide (FE(NO)) was evaluated in children with asthma after 4 to 6 years of treatment with budesonide, nedocromil, or albuterol as needed. STUDY DESIGN: FE(NO), spirometry, total eosinophil count, and serum eosinophil cationic protein levels were obtained from 118 children at the Denver site of the Childhood Asthma Management Program upon completion of treatment and after a 2- to 4-month washout. RESULTS: Budesonide-treated patients had significantly lower median (1st, 3rd quartile) FE(NO) (21.5 [13.2, 84.4] vs 62.5 [26.2, 115.0] ppb, P <.01) and eosinophil cationic protein levels (17.4 [10.1, 24.3] vs 24.0 [15.4, 33.9] mg/dL, P =.05) compared with placebo, whereas no differences were noted between nedocromil and placebo groups. After washout, FE(NO) levels were similar between the three treatments. FE(NO) levels significantly correlated with degree of bronchial hyperresponsiveness, bronchodilator reversibility, allergen skin prick tests, serum IgE, and total eosinophil count. FE(NO) levels were also higher in patients with nocturnal symptoms and in patients requiring beta-agonist use at least once weekly. CONCLUSIONS: Budesonide therapy was more effective than nedocromil in reducing FE(NO). Unfortunately, the effects of long-term budesonide were not sustained after its discontinuation. FE(NO) may be a complementary tool to current practice guidelines in assessing asthma control and medication response. PMID- 12756378 TI - Who is breast-feeding? Recent trends from the pregnancy risk assessment and monitoring system. AB - OBJECTIVE: To examine breast-feeding initiation and continuation among women with recent live births in 10 states. STUDY DESIGN: By using Pregnancy Risk Assessment and Monitoring System surveillance data (n = 96,204), we assessed breast-feeding initiation and continuation for > or =10 weeks among women with recent deliveries from 1993 to 1998. We used 1993 as the base for comparing results by using univariate and multivariate analyses. RESULTS: Ten states showed a significant increase of 18% in initiation of breast-feeding from 1993 to 1998, from 57.0% (95% confidence interval [CI], 55.6-58.4) to 67.5% (95% CI, 66.1-68.9). Initiation increased among vulnerable groups such as low-income and black women, participants in the Special Supplemental Nutrition Program for Women, Infants, and Children program, and mothers of infants admitted to the neonatal intensive care unit. The percentage of women predominantly breast-feeding at > or =10 weeks among women who initiated remained stable: 58.5% (95% CI, 56.5-60.5) in 1993 and 57.9% (95% CI, 56.0-59.8) in 1998. More women in vulnerable groups initiated breast-feeding, but those from higher socioeconomic groups continued breast feeding. CONCLUSIONS: Breast-feeding initiation significantly increased, and several states exceeded the year 2010 objective. Breast-feeding continuation among women who initiated remained stable; however, gaps remained, indicating a continued need to implement breast-feeding promotion programs. PMID- 12756379 TI - Intrapartum antibiotics and neonatal invasive infections caused by organisms other than group B streptococcus. AB - OBJECTIVES/STUDY DESIGN: Administration of group B streptococcal (GBS) antibiotic prophylaxis to women in labor has dramatically reduced the incidence of GBS neonatal disease, but there is little information on its impact on neonatal infections caused by other organisms. We conducted a nested case-control study to define the association between maternal intrapartum antibiotics and risk of neonatal non-GBS infection. RESULTS: In our study population, 114 of 13,224 infants had 115 non-GBS infections. The incidence of non-GBS neonatal infections fell during the study period, ranging from an attack rate of 9.6 per 1000 infants in 1990 to 1992 to 8.0 per 1000 infants in 1996 to 1998, although this trend was not statistically significant (P >.05). The unadjusted association between neonatal infection and GBS prophylaxis was 0.89 (95% CI, 0.29, 2.6) and between neonatal infection and maternal intrapartum antibiotic due to any cause was 1.3 (95% CI, 0.65, 2.8). CONCLUSIONS: The current policy of GBS maternal prophylaxis does not appear to convey excess risk of non-GBS infection to neonates. PMID- 12756380 TI - Relation between pain and self-injurious behavior in nonverbal children with severe cognitive impairments. AB - OBJECTIVES: To explore whether self-injurious behavior (SIB) alters pain expression in children with severe cognitive impairments and the relation between SIB and chronic pain. STUDY DESIGN: Caregivers of 101 nonverbal children 3 to 18 years of age (55% boys) completed the Non-Communicating Children's Pain Checklist Revised (NCCPC-R) retrospectively and for an observed pain episode. Caregivers of children with SIB (n = 44) completed the Behavior Problems Inventory, the Self Injury Grid, and the Self-Injury and Self-Restraint Checklist. RESULTS: Multivariate analysis of variance indicated that NCCPC-R scores did not differ between children with and those without SIB. However, t tests indicated that children with chronic pain (n = 13) self-injured less body surface (P =.01) and fewer body sites (P =.04) than did children without (n = 31). Multiple Correspondence Analysis generated 2 dimensions (49% variance), suggesting a distinction between two SIB forms: (1) high frequency of SIB to the head/hand and absence of chronic pain and (2) less frequent SIB near the site of pain. CONCLUSIONS: Children with severe cognitive impairments who display SIB do not have reduced pain expression, and chronic pain may influence the frequency and location of SIB. Further research should examine the usefulness of these findings for management of SIB and pain. PMID- 12756381 TI - Parathyroid function and growth in 22q11.2 deletion syndrome. AB - OBJECTIVE: To determine the frequency and expression of hypoparathyroidism and the factors of short stature in 22q11.2 deletion syndrome to optimize clinical care. STUDY DESIGN: Cross-sectional study of 39 patients 9.7 +/- 0.8 (2.5-20) years of age. RESULTS: The congenital abnormalities were cardiac defects in 33 of 39, thymus hypoplasia in 15 of 18 evaluated, and craniofacial dysmorphy in all; 15 patients (39%) had had one or more seizures. Before evaluation, 12 patients were hypocalcemic, with (n = 4) or without clinical manifestations, diagnosed before 1 month in 10 cases, at 3 months or 12 years in two others. At evaluation, 9 patients were hypocalcemic, 5 of 9 had been hypocalcemic, and 8 others had parathyroid hormone (PTH) concentrations low for their ionized calcium. One had high PTH without hypocalcemia and 2 were hypercalcemic. The values were below -2 SD at birth for weight and/or height in 26% of cases and at evaluation for height and body mass index in 23% and for insulin-like growth factor-I in 37%. CONCLUSIONS: Parathyroid function was abnormal in 27 of 39 (69%) patients. This was not diagnosed in the majority. Short stature was probably due to intrauterine growth restriction, underweight, and growth hormone deficiency, as suggested by low insulin-like growth factor I. PMID- 12756382 TI - Randomized, double-blind, placebo-controlled trial of oral albuterol in infants with mild-to-moderate acute viral bronchiolitis. AB - OBJECTIVE: To determine whether oral albuterol is effective in reducing symptomatology of acute viral bronchiolitis in infants with mild-to-moderate illness. STUDY DESIGN: In a randomized, double-blind trial, previously well infants were randomized upon discharge from the emergency department to receive either albuterol (0.1 mg/kg/dose) three times per day or placebo three times per day for 7 days. Daily standardized telephone interviews were conducted for as long as 14 days. The primary outcome was the time to resolution of illness. Secondary outcomes included time to normal feeding, normal sleeping, quiet breathing, resolved cough, and coryza. RESULTS: We studied 129 infants (albuterol, n = 64; placebo, n = 65). Baseline characteristics were similar between groups. The overall mean age was 5.3 months, 60% were male, and 49 of 61 tested infants were positive for respiratory syncytial virus. The median (95% confidence interval) time to resolution of illness (days) was similar: albuterol, 9.0 (8-13); placebo, 8.0 (7-9); P =.3) (log-rank test). There were no significant group differences in any secondary outcome. Health care revisit and admission rates were similar between groups. CONCLUSIONS: No significant group differences in either primary or secondary outcomes in infants treated with oral albuterol versus placebo were found. The widespread use of oral albuterol in this patient group is not recommended. PMID- 12756383 TI - Enuresis in children with sleep apnea. AB - OBJECTIVES: To test the hypothesis that the presence of nocturnal enuresis is related to the severity of sleep apnea, we examined the relation between the Respiratory Disturbance Index (RDI, apneas plus hypopneas per hour of sleep) and the presence and severity of enuresis. STUDY DESIGN: All children 4 years of age and older who were referred to our sleep center for suspected sleep disordered breathing (SDB) were asked whether and how frequently they currently wet the bed. All patients underwent full overnight polysomnography (PSG). The relation between RDI and enuresis was examined by chi(2) analysis. A value of P <.05 was considered statistically significant. RESULTS: Ninety boys and 70 girls were studied; 66 children (41%) described current enuresis. At all ages, enuresis was more prevalent in our patients than control patients in the literature. Children with an RDI of < or =1 had a significantly lower prevalence of enuresis (17%) than did children with an RDI >1 (47%) (P <.05). Fourteen percent of children with an RDI < or =1 had frequent enuresis, compared with 32% of children with an RDI >1 (P <.05). There was no significant difference in the prevalence of enuresis in children with an RDI 1 to 5, 5 to 15, or >15 (P =.92). CONCLUSIONS: There is a high prevalence of enuresis in children with suspected sleep disordered breathing. Children with an RDI >1 were at higher risk for enuresis than children with an RDI < or =1. This may be due to the effects of obstructive sleep apnea on arousal response, bladder pressure, or urinary hormone secretion. PMID- 12756384 TI - Umbilical cord blood transplantation in Wiskott Aldrich syndrome. AB - OBJECTIVE: To report the use of umbilical cord blood (UCB) stem cell transplantation in Wiskott Aldrich syndrome (WAS) when a matched sibling donor was unavailable. METHODS: Three children with WAS received unrelated umbilical cord blood stem cell transplantation after a preparative regimen for the treatment of combined immunodeficiency diseases. The patients ranged in age from 1.9 to 7.9 years. The cord blood units were 4/6 HLA antigen matches in 2 children and 5/6 in 1 child, with molecular HLA-DR match in all 3 children. RESULTS: The time for neutrophil engraftment (ANC >500/mm(3)) was 11 to 16 days, and the average time for platelet engraftment was 36 to 49 days. One patient had no evidence of GvHD, 1 patient grade I, and 1 patient grade II. No patient had chronic GvHD. The patient with grade II GvHD also had gut involvement. Immunologic reconstitution demonstrated that cord blood stem cell transplantation resulted in consistent and stable T-, B-, and NK-cell development. Functional B cell antibody responses revealed that 2 of the patients in whom IVIG has been discontinued had low detectable antibody responses to tetanus and diphtheria toxoid immunizations at 18 to 24 months after transplantation. CONCLUSIONS: Unrelated umbilical donor cord blood is an alternative source of stem cells for transplantation in children with WAS when a suitable HLA-matched donor is not available. Benefits of UCB include rapid and reliable recovery of immune function, low risk of GvHD, and low viral transmission rate. PMID- 12756385 TI - Red cell and plasma plant sterols are related during consumption of plant stanol and sterol ester spreads in children with hypercholesterolemia. AB - OBJECTIVE: To show whether the ratios of squalene and cholesterol precursor sterols to cholesterol and cholestanol and plant sterols to cholesterol change differently in plasma and especially in the red cells of hypercholesterolemic children during consumption of plant stanol and sterol ester spreads. STUDY DESIGN: In a randomized, double-blind, crossover study, hypercholesterolemic children (n = 23) consumed low-fat plant stanol and sterol ester spreads for 5 week periods separated by a 5-week washout period. Plasma and red cell lipids, squalene, and noncholesterol sterols were measured before and at the end of each period. RESULTS: The plant stanol and sterol ester spreads lowered plasma total ( 9% and -6%, respectively) and low-density lipoprotein (-12% and -9%) cholesterol but had no effect on red cell cholesterol, high-density lipoprotein cholesterol, or plasma triglycerides. The ratios of plasma and red cell sitosterol and campesterol to cholesterol decreased by 32% to 36% (P <.001) with the plant stanol ester and increased by 40% to 52% (P <.001) with the sterol ester spread. CONCLUSIONS: Consumption of plant sterols increases and consumption of plant stanols decreases the ratios of plant sterols to cholesterol in red cells of hypercholesterolemic children proportionately to the respective changes in plasma. PMID- 12756386 TI - Is McCune-Albright syndrome overlooked in subjects with fibrous dysplasia of bone? AB - OBJECTIVE: McCune-Albright syndrome (MAS) is characterized by a clinical triad of endocrinopathies, cafe au lait pigmentation, and polyostotic fibrous dysplasia of bone. We hypothesized that children diagnosed with fibrous dysplasia are not routinely being evaluated for coexisting endocrine dysfunction or MAS. Our objective was to prospectively screen subjects with fibrous dysplasia for endocrine disease and G(s)alpha gene (GNAS1 )-activating mutations. STUDY DESIGN: Nine subjects who presented with fibrous dysplasia and were followed in orthopedic clinics were evaluated for other manifestations of MAS. Genomic DNA was isolated from blood, and mutation analysis of GNAS1 was performed. RESULTS: On physical examination, 5 of 9 subjects were found to have cafe au lait pigmentation. Three of 9 subjects had TSH levels below the normal range. One of these subjects was found to have hyperthyroidism and was treated by total thyroidectomy. GNAS1 mutations were identified in 5 of 9 subjects with either monostotic or polyostotic fibrous dysplasia of bone. CONCLUSIONS: We conclude that a substantial proportion of children being followed for fibrous dysplasia of bone have unrecognized clinical and laboratory features of MAS. These children are at risk for endocrinopathy and should be screened accordingly. PMID- 12756387 TI - Adverse events with rhGH treatment of patients with chronic renal insufficiency and end-stage renal disease. AB - OBJECTIVE: Recombinant human growth hormone (rhGH) has been used to improve the growth retardation associated with chronic renal insufficiency (CRI) and end stage renal disease. We determined the incidence of one of four targeted adverse events (AEs): malignancy, slipped capital femoral epiphysis (SCFE), avascular necrosis (AN), and intracranial hypertension (ICH). STUDY DESIGN: During a 6.5 year period, we prospectively assessed patients enrolled in the CRI, dialysis, and transplant registries of the North American Renal Transplant Cooperative Study. The availability of an untreated control population facilitated determining whether or not there was the association between the AE and rhGH treatment. RESULTS: Of the targeted AE, the only significant relation with rhGH treatment was the presence of ICH in patients with CRI; however, in all 3 instances, ICH occurred 2, 50, and 1131 days after discontinuation of rhGH. Considering that the mechanism of ICH in rhGH-treated patients is thought to be increased CSF production, rhGH probably had no role in the development of ICH in at least 2 of the 3 patients with CRI. A number of nontargeted AE were identified that have been associated with rhGH treatment in patients without renal disease. The incidence of glucose intolerance, pancreatitis, progressive deterioration of renal function, acute allograft rejection, and fluid retention were not more frequent in those receiving rhGH treatment compared with the control population. CONCLUSIONS: This report validates the importance of a control population in ascribing AE to any therapeutic intervention. Previously identified AE associated with rhGH treatment are infrequent in patients with CRI and end-stage renal disease. PMID- 12756388 TI - Does age play a role in recovery from sports-related concussion? A comparison of high school and collegiate athletes. AB - OBJECTIVE: To evaluate symptoms and neurocognitive recovery patterns after sports related concussion in high school and college athletes. STUDY DESIGN: College athletes (n = 371) and high school athletes (n = 183) underwent baseline neuropsychological evaluation between 1997 and 2000. Individuals who received a concussion during athletic competition (n = 54) underwent serial neuropsychologic evaluation after injury and were compared with a noninjured within-sample control group (n = 38). Main outcome measures included structured interview, four memory measures, and Concussion Symptom Scale ratings. Baseline to postinjury change scores and multiple analyses of variance were used to compare recovery curves within and between groups. RESULTS: High school athletes with concussion had prolonged memory dysfunction compared with college athletes with concussion. High school athletes performed significantly worse than age-matched control subjects at 7 days after injury (F = 2.90; P <.005). College athletes, despite having more severe in-season concussions, displayed commensurate performance with matched control subjects by day 3 after concussion. Self-report of postconcussion symptoms by student athletes was not predictive of poor performance on neuropsychologic testing. CONCLUSIONS: Caution and systematic evaluation should be undertaken before returning athletes with concussion to competition. Sole reliance on the self-report of the athlete may be inadequate. Preliminary data may suggest a more protracted recovery from concussion in high school athletes. PMID- 12756390 TI - A 16-year-old boy with purpura and leg pain. PMID- 12756389 TI - Parents' attitudes to children's participation in randomized controlled trials. AB - OBJECTIVE: To explore parents' attitudes to children's participation in randomized controlled trials. STUDY DESIGN: Qualitative analysis of focus group discussions involving 33 parents from a pediatric teaching hospital and local school in Australia. Parents varied in age, gender, ethnicity, level of education, research experience, and child's health status. The transcribed discussions were analyzed by theme linkage using the constant comparative method. RESULTS: Parents balance risks and benefits when deciding about trial participation for their child. Perceived benefits include the offer of hope, better care of their child, the opportunity to access new treatments, healthcare professionals and health information, meeting others in similar circumstances, and helping others. Perceived risks include potential side effects, being randomized to ineffective treatments, and the inconvenience of participation. The decision for trial participation is also influenced by parental factors (parents' knowledge, beliefs, and emotional responses), child factors (the child's health status and preference about participation), trial factors (the use of placebos and uncertainties of participation), and doctor factors (doctor's recommendations and communication of trial information). CONCLUSIONS: Educating parents about trials, improving communication between trialists, pediatricians, and parents, increasing incentives while decreasing inconveniences, and providing decision aids for parents may increase parents' willingness to participate in trials. PMID- 12756391 TI - Time to positive culture results in neonatal Candida septicemia. AB - To determine the time required for blood cultures to be detected as positive in neonates with invasive Candida infection, we analyzed 207 positive culture episodes in 74 infants. Time to positive result was 37 +/- 14 hours, and 97% of infected neonates were detected by 72 hours if not exposed to antifungal therapy. PMID- 12756392 TI - Cholangitis after hepatic portoenterostomy for biliary atresia: a multivariate analysis of risk factors. AB - Cholangitis occurred in 59% of 77 patients who underwent the portoenterostomy procedure for biliary atresia between 1980 and 2000. Good postoperative bile drainage was associated with a lower risk of cholangitis than partial (odds ratio, 5.72; 95% CI, 2.89-11.3) or poor (odds ratio, 3.29; 95% CI, 1.89-5.7) bile drainage. Cholangitis was not an independent risk factor for death or liver transplantation. PMID- 12756393 TI - Digoxin-carvedilol interactions in children. AB - Digoxin is often coadministered with carvedilol in children with severe ventricular failure. In eight children (age 2 weeks to 8 years), the oral clearance of digoxin decreased by half with carvedilol, and two of them had digoxin toxicity. Carvedilol increases serum concentrations of digoxin in children, and its dose may need to be reduced to avoid toxicity. PMID- 12756394 TI - Electrophysiologic evidence of impaired cross-modal recognition memory in 8-month old infants of diabetic mothers. AB - OBJECTIVES: Previous studies have shown deficits in auditory and visual recognition memory in infants of diabetic mothers. The purpose of this study was to further investigate memory development in infants of diabetic mothers (IDMs) by evaluating cross-modal recognition memory followed by behavioral memory testing at 8 months of age. STUDY DESIGN: Cross-modal (tactile to vision) recognition memory was evaluated using event related potentials. Control and IDMs palpated an object without seeing it and were then tested on their ability to recognize that object visually. Infants were then tested behaviorally on their ability to recognize and discriminate faces. The Bayley Scales of Infant Development was administered at 12 months of age. RESULTS: Control infants showed typical event-related potential patterns indicative of intact cross-modal recognition memory, whereas the IDMs did not show any evidence of recognition of the palpated object. Neither group showed behavioral evidence of visual recognition memory. Both groups had Bayley scores in the normal range, although controls had slightly higher mental development index scores than IDMs. CONCLUSIONS: IDMs showed neurophysiologic evidence of persistent subtle impairments in hippocampally-based recognition memory, despite having normal one year developmental scores. PMID- 12756395 TI - Glutathione metabolism and antioxidant enzymes in children with Down syndrome. AB - Oxidative stress has been proposed as a pathogenic mechanism of atherosclerosis, cell aging, and neurologic disorders in Down syndrome. This study demonstrates a systemic decrease of all glutathione forms, including glutathionyl-hemoglobin, in the blood of children with Down syndrome. Furthermore, we obtained a disequilibrium, in vivo, between the antioxidant enzyme activities. PMID- 12756396 TI - Temporary tattoo dermatitis. PMID- 12756397 TI - Evaluating neonatal developmental care. PMID- 12756398 TI - Evaluating neonatal developmental care. PMID- 12756399 TI - No evidence of interaction between PCBs and critical developmental factors. PMID- 12756400 TI - Nitric oxide in Kawasaki disease. PMID- 12756401 TI - Insulin resistance and vascular remodelling, in relation to left ventricular mass, geometry and function: an answer to LIFE? PMID- 12756403 TI - Red blood cell K+ could be a marker of K+ changes in other cells involved in blood pressure regulation. AB - The aim of this study is to determine whether red blood cell K(+) content (RBC(Ki)) is associated with blood pressure levels and, if so, could RBC(Ki) be a marker of potassium changes in other cells involved in blood pressure regulation. The study was performed on 50 untreated hypertensives, 32 of their offspring and 50 age- and sex-matched controls. Systolic (SBP) and diastolic (DBP) blood pressures, height, weight, plasma, urine and red blood cell electrolytes were measured in all subjects. RBC(Ki) was significantly lower in hypertensives than in offspring of hypertensives and normotensive controls. Offspring of hypertensives had significantly lower RBC(Ki) than normotensive controls. Plasma K(+) was significantly lower both in hypertensives and offspring of hypertensives when compared to normotensive controls. A significant negative correlation was found in hypertensives between RBC(Ki) and DBP (r=-0.27, P=0.04) and in offspring of hypertensives between RBC(Ki) and DBP (r=-0.43, P=0.02). A significant correlation was found in hypertensives between RBC(Ki) and plasma K(+) (r=0.3, P=0.02). A positive correlation with borderline significance was found in hypertensives between RBC(Ki) and ionized Ca(2+) (r=0.2, P=0.1). In conclusion, our results support the hypothesis that RBC(Ki) is associated with blood pressure levels and that the measurement of RBC(Ki) levels may represent a biochemical marker for K(+) changes in other cells involved in blood pressure regulation. Further studies are necessary to explain the exact mechanisms of reduced RBC(Ki) levels in hypertensive patients and their offspring. PMID- 12756404 TI - Physical mapping of autonomic/sympathetic candidate genetic loci for hypertension in the human genome: a somatic cell radiation hybrid library approach. AB - Allelic variation at multiple genetic loci may contribute to hypertension. Since autonomic/sympathetic dysfunction may play an early, pathogenic, heritable role in hypertension, we evaluated candidate loci likely to contribute to such dysfunction, including catecholamine biosynthetic enzymes, catecholamine transporters, neuropeptides, and adrenergic receptors. Since chromosomal locations and physical map positions of many of these loci had not yet been identified, we used the GeneBridge4 human/hamster radiation (somatic cell) hybrid library panel (resolution approximately 1 to approximately 1.5 Mb), along with specifically designed oligonucleotide primers and PCR (200-400 bp products) to position these loci in the human genome. Primers were designed from sequences outside the coding regions (3'-flanking or intronic segments) to avoid cross species (hamster) amplification. Chromosomal positions were assigned in cR (centi Ray) units ( approximately 270 Kbp/cR(3000) for GeneBridge 4). A total of 13 loci were newly assigned chromosomal positions; of particular interest was a cluster of adrenergic candidate loci on chromosome 5q (including ADRB2, ADRA1A, DRD1, GPRK6, and NPY6R), a region harbouring linkage peaks for blood pressure. Such physical map positions will enable more precise selection of polymorphic microsatellite and single nucleotide polymorphism markers at these loci, to aid in linkage and association studies of autonomic/sympathetic dysfunction in human hypertension. PMID- 12756402 TI - Are left ventricular mass, geometry and function related to vascular changes and/or insulin resistance in long-standing hypertension? ICARUS: a LIFE substudy. AB - Vascular hypertrophy and insulin resistance have been associated with abnormal left ventricular (LV) geometry in population studies. We wanted to investigate the influence of vascular hypertrophy and insulin resistance on LV hypertrophy and its function in patients with hypertension. In 89 patients with essential hypertension and electrocardiographic LV hypertrophy, we measured blood pressure; insulin sensitivity by hyperinsulinaemic euglucaemic clamp; minimal forearm vascular resistance (MFVR) by plethysmography; intima-media cross-sectional area of the common carotid arteries (IMA) by ultrasound; and LV mass, relative wall thickness (RWT), systolic function and diastolic filling by echocardiography after two weeks of placebo treatment. LV mass index correlated to IMA/height (r=0.36, P=0.001), serum insulin (r=-0.25, P<0.05), plasma glucose (r=-0.34, P<0.01), and showed a tendency towards a correlation to insulin sensitivity (r=0.21, P=0.051), but was unrelated to MFVR. Deceleration time of early diastolic transmitral flow positively correlated to IMA/height (r=0.30, P<0.01). The ratio between early and atrial LV filling peak flow velocity negatively correlated to MFVR(men) (r=-0.30, P<0.05). Endocardial and midwall systolic LV function were not related to vascular hypertrophy, plasma glucose, serum insulin or insulin sensitivity. In conclusion, insulin resistance was not related to LV hypertrophy or reduced LV function. However, high thickness of the common carotid arteries was associated with LV hypertrophy and high deceleration time of early diastolic transmitral flow. High MFVR was associated with low ratio between early and atrial LV filling peak flow velocity. This may suggest that systemic vascular hypertrophy contributes to abnormal diastolic LV relaxation in patients with hypertension and electrocardiographic LV hypertrophy. PMID- 12756405 TI - Ambulatory blood pressure and left ventricular structure and function in relation to the G-protein beta3-subunit polymorphism C825T in White Europeans. AB - The 825T allele of the G-protein beta(3)-subunit is associated with increased intracellular signalling. Its association with hypertension is inconsistent. We, therefore, studied the C825T polymorphism in relation to ambulatory blood pressure as well as left ventricular structure and function in two European populations. We genotyped 248 parents and 318 offspring, enrolled in the European Project on Genes in Hypertension in Cracow, Poland (n=286) and in Novosibirsk, Russian Federation (n=280). The 24-h ambulatory blood pressure was recorded using oscillometric SpaceLabs 90207 monitors. Within each centre, a single observer performed two-dimensionally guided M-mode echocardiography and Doppler sonography to measure left ventricular structure (American Society of Echocardiography conventions) and diastolic function: early (E) and late (A) peak diastolic inflow velocities. We used analysis of covariance and generalized estimating equations to allow for covariables and nonindependence among related subjects. Genotype frequencies were similar (P=0.25) in Cracow and Novosibirsk and amounted to 44.7% for CC, 47.2% for CT, and 8.1% for TT. Among parents (mean age: 51.3 years)-but not among offspring (mean age 25.1 years)-24-h, daytime and night time systolic blood pressures were 5-6 mmHg higher in TT homozygotes than in C allele carriers. In TT homozygous parents (-8.2 cm/sec, P=0.004) as well as in TT homozygous offspring (-7.5 cm/sec, P=0.02), the E-wave was significantly reduced, which in offspring also resulted in a lower E/A ratio (-0.25, P=0.002). Neither in parents nor in offspring, left ventricular mass index was associated with the C825T polymorphism. In conclusion, in TT homozygotes of both generations, early left ventricular relaxation was reduced. In TT homozygous parents, the latter observation might be because of the higher systolic pressure associated with the TT genotype. PMID- 12756406 TI - Exercise blood pressure response is related to left ventricular mass. AB - An exaggerated SBP response to exercise has been associated with increased left ventricular (LV) mass in some but not all studies. A total of 43 women and 34 men, aged 55-75 years, without evidence of cardiovascular disease, with a mean resting BP of 142+/-9/77+/-8 mmHg had their BP measured at rest and during maximal treadmill exercise. LV mass was measured using magnetic resonance imaging. LV mass was adjusted for lean body mass, which was assessed by dual energy X-ray absorptiometry. LV mass was within the normal range for the majority of the subjects. Among the resting and exercise BP indices, maximal SBP was the strongest correlate of LV mass (r=0.41, P<0.05). In multivariate analysis, maximal SBP was independently associated with LV mass after adjustment for lean body mass and gender, explaining 3% of the variance (P<0.05). Maximal exercise SBP is a modest but still independent predictor of LV mass in older persons with normal LV mass. These results raise the possibility that the SBP response to maximal exercise is an early marker of LV hypertrophy. PMID- 12756408 TI - Prevalence of primary hyperaldosteronism in moderate to severe hypertension in the Central Europe region. AB - Recently published studies from different parts of the world report significantly higher prevalence of primary hyperaldosteronism (PH) in hypertensives (ranging from 5 to 25%) than the previously accepted figures. There have been no data so far about the prevalence of PH in Central Europe. Therefore, we have undertaken this study to evaluate the prevalence of PH in patients with moderate to severe hypertension referred to a hypertension unit in the Czech Republic, together with the determination of the percentage of different subtypes of PH including familial forms. In addition to that, we have evaluated the prevalence of other types of secondary forms of hypertension.A total of 402 consecutive patients (230 females and 172 males) with hypertension, referred to our hypertension unit, were studied. Positive aldosterone/renin ratio (ARR, (ng/100 ml)/(ng/ml/h)) >/=50 as a more strict marker of PH was found in 87 patients (21.6%), 30% of them were normokalaemic. The diagnosis of PH was later confirmed in 77 cases (89%); the total prevalence of PH was thus 19%. PH consisted of the following forms: idiopathic hyperaldosteronism 42%, unilateral aldosterone-producing adenoma 36%, unilateral hyperplasia 7%, nonclassifiable PH (refused operation/adrenal venous sampling) 13%, familial hyperaldosteronism type 1.2%. The prevalence of other types of secondary hypertension was as follows: pheochromocytoma 5%, renovascular 4.5%, hypercortisolism 2%, renal 0.75%. In conclusion, we have noted that PH in the Central Europe region (Czech Republic) is the most frequent form of endocrine hypertension with a considerably high prevalence in moderate to severe hypertension. Application of more strict criteria raises the probability of correct diagnosis of PH including the early normokalaemic stages of PH. PMID- 12756407 TI - Factor analysis of possible risks for hypertension in a black South African population. AB - To date only a small number of studies have investigated the pattern of associations within a set of hypertension risks. The objective of this study was therefore to examine the interrelation of main hypertension risks in an African population by using factor analysis in order to detect underlying risk patterns. Subjects aged 16-70 years (N=963) were recruited from 37 randomly selected sites throughout the North West Province during 1996-1998. Exclusion criteria were pregnancy, lactation, casual visitors, drunkenness and treatment for chronic diseases, such as hypertension. Subjects with blood pressures exceeding 140/90 mmHg were classified as hypertensive. Children aged 10-15 years were also recruited from 30 randomly selected schools during 2000-2001 (N=694). Children were classified as hypertensive when an average systolic or diastolic blood pressure greater than or equal to the 90th percentile for age and sex was encountered, while correcting for height. The following hypertension risks were measured: urbanisation, obesity, plasma fibrinogen, lipids, insulin, serum gamma glutamyl-transferase, dietary intake, smoking and alcohol consumption. From 23 risks the factor analysis disclosed five factors that explained 56.2% of the variance in the male and 43.5% of the variance in the female group: an urban malnutritional phenomenon, the metabolic syndrome X, a hypercholesterolaemic and obesity complex, an alcoholic hypertriglyceridaemia, and central and peripheral cardiovascular hypertensive effects. In conclusion, South Africans migrating from rural to urban areas adapt to a new lifestyle with numerous risks, resulting in conditions like malnutrition, the metabolic syndrome X, dyslipidaemia, alcoholism, obesity and increased peripheral vascular resistance. For successful prevention of hypertension in a population in transition, a whole risk pattern should be corrected, rather than an individual risk by implementing lifestyle modification programmes. PMID- 12756409 TI - Mean arterial pressure and pulse pressure are associated with different clinical parameters in chronic haemodialysis patients. AB - The mean arterial pressure (MAP) usually serves as an expression of blood pressure in patients on chronic haemodialysis (PCHD), instead of using solely systolic or diastolic pressure. Pulse pressure (PP) has been recognized as an important correlate of mortality in PCHD. We conducted this study in order to demonstrate clinical and biochemical determinants and variability of predialysis and postdialysis MAP and PP values. A total of 136 single haemodialysis (HD) treatments in 23 subjects (PCHD, 11 male and 12 female patients) were processed during 15 months. MAP before HD was in negative correlation with haemoglobin (P<0.001) and body mass index (BMI) (P<0.001), and in positive correlation with weekly erythropoietin dosage (P=0.017). MAP after HD was in negative correlation with haemoglobin (P<0.001), ultrafiltration per HD (P=0.015), and BMI (P=0.001), and in positive correlation with weekly erythropoietin dosage (P=0.003). PP before HD was in negative correlation with parathyroid hormone (PTH) level (P=0.020), haemoglobin (P<0.001), ultrafiltration per HD (P=0.001), and years on the chronic HD treatment (P=0.001), and in positive correlation with weekly erythropoietin dosage (P<0.001) and age (P<0.001). PP after HD was in significant negative correlation with PTH (P=0.015), haemoglobin (P=0.005), ultrafiltration per HD (P<0.001), BMI (P=0.003), and in positive correlation with weekly erythropoietin dosage (P<0.001) and age (P=0.004). Multiple regression analyses unveiled the strongest and negative correlations between MAP before HD and BMI (beta=-0.37, P=0.01); MAP after HD and haemoglobin (beta=-0.36, P=0.01); PP after HD and ultrafiltration/body weight ratio (beta=-0.41, P<0.001). The strongest and positive correlation was found between PP before HD and erythropoietin dosage per week (beta=0.51, P&<0.001). In conclusion, our findings support the assumption that PP and MAP are associated with different clinical parameters. PP values have advantages as the method of blood pressure expression. PMID- 12756410 TI - Hyperhomocysteinemia but not MTHFR genotype is associated with young-onset essential hypertension. PMID- 12756411 TI - Abdominal adiposity, age, education level and therapy associated with uncontrolled hypertension. PMID- 12756412 TI - Virology and immunology of gene therapy, or virology and immunology of high MOI infection with defective viruses. PMID- 12756413 TI - Molecular basis of the inflammatory response to adenovirus vectors. AB - Adenovirus vectors are extensively studied in experimental and clinical models as agents for gene therapy. Recent generations of helper-dependent adenovirus vectors have the majority of viral genes removed and result in vectors with a large carrying capacity, reduced host adaptive immune responses and improved gene transfer efficiency. Adenovirus vectors, however, activate innate immune responses shortly after administration in vivo. Unlike the adaptive response, the innate response to adenovirus vectors is transcription independent and is caused by the viral particle or capsid. This response results in inflammation of transduced tissues and substantial loss of vector genomes in the first 24 h. The adenovirus capsid activates a number of signaling pathways following cell entry including p38 mitogen-activated protein kinase and extracellular signal-regulated kinase (ERK) that ultimately lead to expression of proinflammatory genes. Various cytokines, chemokines and leukocyte adhesion molecules are induced by the adenovirus particle in a wide range of cell types providing a molecular basis for the inflammatory properties of these vectors. An understanding of the innate response to adenovirus vectors is essential to overcome the last remaining hurdle to improve the safety and effectiveness of these agents. PMID- 12756414 TI - Immune responses to replication-defective HSV-1 type vectors within the CNS: implications for gene therapy. AB - Herpes simplex virus (HSV) is a naturally occurring double-stranded DNA virus that has been adapted into an efficient vector for in vivo gene transfer. HSV based vectors exhibit wide tropism, large transgene size capacity, and moderately prolonged transgene expression profiles. Clinical implementation of HSV vector based gene therapy for prevention and/or amelioration of human diseases eventually will be realized, but inherently this goal presents a series of significant challenges, one of which relates to issues of immune system involvement. Few experimental reports have detailed HSV vector-engendered immune responses and subsequent resolution events primarily within the confines of the central nervous system. Herein, we describe the immunobiology of HSV and its derived vector platforms, thus providing an initiation point from where to propose requisite experimental investigation and potential approaches to prevent and/or counter adverse antivector immune responses. PMID- 12756416 TI - Immunity to adenovirus and adeno-associated viral vectors: implications for gene therapy. AB - Viral vectors have provided effective methods for in vivo gene delivery for therapeutic purposes. The ability of viruses to infect a wide variety of cell types in vivo has been exploited for several applications, such as liver, lung, muscle, brain, eye and many others. Immune responses directed towards the viral capsids and the transgene products have severely affected the ability of these vectors to induce long-term gene expression. This paper reviews the influence of viral vectors on antigen-presenting cells (APC), which are central to the induction of innate as well as adaptive immune responses. In this respect, we have focused on adenovirus and adeno-associated viruses because of the polar responses these vector systems induce in vivo. While adenovirus vector can induce significant inflammatory responses, adeno-associated viral vectors are characterized by their inability to consistantly induce immune responses to the transgene product. Understanding the mechanism of infection, transduction and activation of APC by viral vectors will provide strategies to develop safe vectors and prevent immune responses in gene therapies. PMID- 12756415 TI - Inflammation and adaptive immune responses to adenoviral vectors injected into the brain: peculiarities, mechanisms, and consequences. PMID- 12756417 TI - Immune responses to adeno-associated virus and its recombinant vectors. AB - Recombinant adeno-associated virus (rAAV) vectors have emerged as highly promising for use in gene transfer for a variety of reasons, including lack of pathogenicity and wide host range. In addition, all virus-encoded genes have been removed from standard rAAV vectors, resulting in their comparatively low intrinsic immunogenicity. For gene replacement strategies, transgenes encoded by rAAV vectors may induce less robust host immune responses than other vectors in vivo. However, under appropriate conditions, host immune responses can be generated against rAAV-encoded transgenes, raising the potential for their use in vaccine development. In this review, we summarize current understanding of the generation of both undesirable and beneficial host immune responses directed against rAAV and encoded transgenes, and how they might be exploited for optimal use of this promising vector system. PMID- 12756418 TI - Immune response following intraocular delivery of recombinant viral vectors. AB - There has been significant progress in the last few years in demonstrating the utility of recombinant viral vectors in treating a variety of ocular diseases. The field has moved beyond 'proof-of-principle' and, in fact, has entered the phase where some of these vectors/paradigms are being or soon will be evaluated in human clinical trials. For this reason and also, to increase the understanding of immunological effects of transgenes/viral vectors on the eye, it is important to summarize what is known about these effects. Here, the biology of and immune responses to intraocular injection of three different recombinant viral vectors - adenovirus, adeno-associated virus (AAV), and lentivirus - are summarized. Perhaps, in part because of the unique immunological environment of the eye, the immunological effects of these viruses appear to be fairly benign. Nevertheless, a significant cell-mediated immune response can develop after intraocular administration of adenovirus. The magnitude of this response is affected by the nature of the intraocular compartment to which this virus is administered. Neither AAV nor lentivirus, however, elicit a cell-mediated response and are thus promising vectors for treatment of chronic ocular (retinal) diseases. PMID- 12756419 TI - Effects of innate immunity on herpes simplex virus and its ability to kill tumor cells. AB - Several clinical trials have or are being performed testing the safety and efficacy of different strains of oncolytic viruses (OV) for malignant cancers. OVs represent either naturally occurring or genetically engineered strains of viruses that exhibit relatively selective replication in tumor cells. Several types of OV have been derived from herpes simplex virus 1 (HSV1). Tumor oncolysis depends on the processes of initial OV infection of tumor, followed by subsequent propagation of OV within the tumor itself. The role of the immune responses in these processes has not been extensively studied. On the contrary, effects of the immune response on the processes of wild-type HSV1 infection and propagation in the central nervous system have been studied and described in detail. The first line of defense against a wild-type HSV1 infection in both naive and immunized individuals is provided by innate humoral (complement, cytokines, chemokines) and cellular (macrophages, neutrophils, NK cells, gammadelta T cells, and interferon producing cells) responses. These orchestrate the lysis of virions and virus infected cells as well as provide a link to effective adaptive immunity. The role of innate defenses in curtailing the oncolytic effect of genetically engineered HSV has only recently been studied, but several of the same host responses appear to be operative in limiting anticancer effects by the replicating virus. The importance of this knowledge lies in finding avenues to modulate such initial innate responses, in order to allow for increased oncolysis of tumors while minimizing host toxicity. PMID- 12756420 TI - Adaptive and innate immune responses to gene transfer vectors: role of cytokines and chemokines in vector function. PMID- 12756421 TI - SARS challenges therapeutics. PMID- 12756423 TI - Drug-herb interaction among commonly used conventional medicines: a compendium for health care professionals. AB - The objective of the review was to consolidate the clinical and pharmacologic aspects of drug-herb interactions to develop a compendium of information to provide prescribers with a measure of the risk of interactions, a description of the clinical consequences, and an assessment of the quality (ie, validity) of evidence. A variety of electronic databases and hand-searched references were used to identify documentation of interactions between herbal products and drugs from the most commonly used therapeutic classes. MEDLINE, Allied and Complementary Medicine Database, CINHAL, HealthSTAR, and EMBASE were searched from 1966 to the present. One hundred sixty-two citations were identified. Only 22 citations met the inclusion criteria. Using a matrix of 165 possible drug-herb interaction pairs (15 therapeutic drug classes by 11 herbal products), we identified 51 (31%) interactions discussed in the literature. Twenty-two of these 51 drug-herb pairs (43%) were supported by randomized clinical trials, case control studies, cohort studies, case series, or case studies. The remaining interaction pairs reflected theoretic reasoning in the absence of clinical data. Most interactions were pharmacokinetic, with most actually or theoretically affecting the metabolism of the affected product by way of the cytochrome p450 enzymes. In this review, warfarin was the most common drug and St. John's wort was the most common herbal product reported in drug-herb interactions. To create a comprehensive and valid list of herb-drug interactions would require a substantial increase in research activities in this area. Improvements in the quality of methodology used are also necessary. PMID- 12756424 TI - Clinical effects of locally delivered nicotine in obstructive sleep apnea syndrome. AB - Obstructive sleep apnea (OSA) is a common disorder characterized by disordered breathing and associated with increased mortality and cardiovascular morbidity. A factor in the pathogenesis of OSA is hypotonia of the upper airway muscles during sleep, resulting in occlusion of the upper airway. Nicotine may be a suitable drug because it is a stimulant of breathing and activity of oropharyngeal muscles. A novel delivery system, a nicotine tooth patch (NTP) that releases nicotine continuously, has been developed by Perio Products (Jerusalem, Israel). A 2-mg NTP achieved low plasma levels of nicotine with high saliva levels (62 microg/mL), presumably resulting in high nicotine levels in the oropharynx. The aim of this study was to evaluate the effect of two doses of NTP, 2 mg and 4 mg, on the clinical features in OSA. Ten subjects with OSA were admitted overnight and monitored by polysomnography at baseline and during two treatments. The treatments were blind and in a randomized order. After a 4.3-mg NTP, T(max) was 40 +/- 16 minutes, C(max) was 123 +/- 43 microg/mL, and terminal T(1/2) was 29 +/ 11 minutes in saliva. Substantial nicotine levels persisted in saliva for approximately 4 hours. There was no effect of nicotine on the apnea-hypopnea index, even during the first 4 hours when there were high levels of nicotine in saliva (26.4 +/- 11.6, 26.8 +/- 19.5, and 26.8 +/- 23), or on sleep stages. Eppworth Sleepiness Scale scores were lower with a 4.3-mg NTP (9.1 +/- 4.5, 9.1 +/- 7.7, and 5.9 +/- 6.5). Locally delivered nicotine at the doses used had no significant effect on OSA. PMID- 12756425 TI - Tender points as predictors of distress and the pharmacologic management of fibromyalgia syndrome. AB - The object of this study was to determine the association between tender point pain ratings, tender point counts and distress in people with fibromyalgia and to review the pharmacotherapy of fibromyalgia. Demographic, psychosocial, and health status information was collected from 316 health maintenance organization members with fibromyalgia. A manual tender point exam was conducted. Tender point counts predicted 3.0%, and tender point severity ratings predicted 8.3%, of the variance in distress. Little difference was found between the variance predicted for physical versus psychologic distress. A principal components analysis of all measures produced four distinct factors: global-physical functioning, tender points, psychologic, and physical. Tender point pain ratings and counts predicted a small but significant amount of variance in distress. In addition, FMS involves at least four rather distinct factors, one of which is related to tender points. Pharmacotherapeutic management is provided on a patient-specific basis including pharmacokinetics, pharmacodynamic, pathophysiologic, and psychosocial needs designed and modulated for each individual patient. PMID- 12756426 TI - Viral respiratory infections due to rhinoviruses: current knowledge, new developments. AB - Viral respiratory infections (VRIs) are among the most common reasons for which primary care providers are consulted. VRIs due to rhinoviruses-the most commonly implicated etiologic agent-constitute a syndrome characterized by signs and symptoms of a cold. Rhinoviruses have been implicated in respiratory tract illnesses such as sinusitis and otitis media, as well as lower respiratory complications in high-risk populations. Most patients treat VRI with over-the counter remedies that have been demonstrated to produce marginal clinical benefits. The development of novel antiviral agents has intensified interest in VRIs. Pleconaril, a capsid-function inhibitor currently under FDA review, has been shown in clinical trials to reduce the duration and severity of rhinovirus VRIs. By targeting the cause of illness, antiviral agents represent an opportunity to reduce the substantial clinical burden of VRI. Furthermore, effective therapies can potentially reduce inappropriate antibiotic use for viral infections. PMID- 12756428 TI - Role of leptin in the regulation of body fluid volume and pressures. AB - Leptin is a circulating polypeptide hormone produced by an adipocyte-specific gene. It regulates energy balance by binding to receptors in the hypothalamus, leading to alterations in food intake, temperature, and energy expenditure. More recent pharmacologic information suggests that this circulating hormone may play an important role in the regulation of body fluid volume and pressures through direct and indirect actions. Although the relevance of the endogenous leptin on cardiovascular and renal function is yet to be clearly determined, it seems to be a potential salt-regulating factor and may function pathophysiologically as a common link to obesity and hypertension. PMID- 12756427 TI - Melanoma chemoprevention: a role for statins or fibrates? AB - Although numerous second-generation isoprenylation inhibitors are proposed or under investigation for the treatment and/or prevention of cancer (eg, R115777, SCH 66336, L-778,123, BMS-214662), the chemotherapeutic and chemopreventive potential of commonly prescribed first-generation isoprenylation inhibitors, the statins, and other classes of lipid-lowering medications, the fibrates, has yet to be seriously explored. Two lipid-lowering medications, lovastatin and gemfibrozil, have been associated with a decreased incidence of melanoma in large, prospective, randomized, double-blind, placebo-controlled clinical cardiology trials. This article reviews melanoma biology and the clinical evidence for the use of lipid-lowering medications for melanoma chemoprevention and/or adjuvant chemotherapy. PMID- 12756429 TI - A swollen joint: why all the fuss? AB - Acute arthritis may be a potential medical emergency. An infected joint causes rapid cartilaginous destruction and risk of future osteoarthritis. Prompt attention to the historical clues and potential causative organisms ensures appropriate therapy. Numerous microbes have been identified as the causative agent in septic arthritis, and various populations have distinct susceptibilities to these specific organisms. One broad classification of septic arthritis is differentiating gonococcal and nongonococcal organisms. This classification is important, as not only do the organisms differ, but the age of the patient and the portal of entry also differ. Aspiration of synovial fluid is paramount for proper diagnosis and management of septic arthritis. In addition, the timely administration of properly chosen antibiotic agents is essential for reducing morbidity and mortality. PMID- 12756430 TI - Severe methemoglobinemia after transesophageal echocardiography. AB - Methemoglobinemia, an increased concentration of methemoglobin in the blood, is an altered state of hemoglobin whereby the ferrous form of iron is oxidized to the ferric state, rendering the heme moiety incapable of carrying oxygen. This can cause hypoxia, cyanosis, or even death. Severe methemoglobinemia resulting from oral benzocaine spray before endoscopic procedures has been reported as a rare complication. We report a case of severe acquired methemoglobinemia resulting from topical benzocaine use before transesophageal echocardiography. This case serves to highlight the severity of methemoglobinemia that can result from an otherwise innocuous agent even in small doses and the fact that prompt recognition and treatment of this disorder can be lifesaving. PMID- 12756431 TI - Pulmonary toxicity during prostate cancer treatment with docetaxel and thalidomide. AB - The standard therapies of surgery, radiotherapy, and hormonal manipulations often fail to control metastatic prostate cancer (PC). Docetaxel and thalidomide may have activity in refractory PC. We highlight the potential pulmonary toxicity when docetaxel is combined with thalidomide. We reviewed three examples of docetaxel and thalidomide pulmonary toxicity at the National Cancer Institute (NCI) and summarized the published literature regarding docetaxel and thalidomide pulmonary toxicity. Docetaxel and thalidomide pulmonary toxicity has the following four main presentations: (1). symptomatic effusions; (2). dyspnea on exertion without any objective pathologic evidence; (3). interstitial lung disease; and (4). pulmonary embolus. As chemotherapy becomes more common in the treatment of PC, clinicians must consider possible pulmonary toxicities. If pulmonary symptoms or signs develop, clinicians should consider holding chemotherapy pending a complete evaluation. PMID- 12756432 TI - Cold-Eeze lozenge for common colds. PMID- 12756434 TI - Vitamin C hypertension. PMID- 12756436 TI - Activated Charcoal RCT. PMID- 12756437 TI - Outcome prediction in terms of functional disability and mortality at 1 year among ICU-admitted severe stroke patients: a prospective epidemiological study in the south of the European Union (Evascan Project, Andalusia, Spain). AB - OBJECTIVES: To analyse 1-year mortality and functional disability outcomes and resource use in critical stroke patients admitted to ICU. DESIGN AND SETTING: Multi-centre, prospective, observational study in 28 Spanish hospitals. PATIENTS. Patients admitted for acute stroke from March-August 1999. INTERVENTION: Collection of data on: severity by Apache III and Glasgow Coma Score; neurological lesion, hospital and 1-year mortality; functional disability at 1 year by Barthel Index and Glasgow Outcome Scale; ICU length of stay, life support techniques, and neurosurgical interventions. MEASUREMENT AND RESULTS: We studied 132 patients: 21% with subarachnoid haemorrhage (SAH), 58% intracerebral haemorrhage (ICH), 20% ischaemic stroke (ISC); Apache III 63+/-29 ICU stay 13+/ 12 days; 74% required mechanical ventilation. Hospital and 1-year mortality was 33% (22%:ISC, 32%:SAH, 37%:ICH) and 53.8% (66%:ISC, 39%:SAH, 54%:ICH), respectively. Age, APACHE III, and diagnosis defined hospital mortality. Age, APACHE III, and Glasgow Coma Score defined 1-year mortality. Barthel Index score improved ( P<0.001) between discharge and 1 year; 73% of patients presented severe disability at discharge vs. 26% at 1 year; 8% minimal/no disability at discharge vs. 43.3% at 1 year. Only 17% of subarachnoid haemorrhage patients presented severe disability at 1 year. Admission Apache III and hospital discharge Barthel Index scores were related to functional outcome at 1 year. CONCLUSIONS: Critical stroke patients are characterized by high severity of illness, elevated resource consumption, and poor outcomes that are mainly influenced by severity and age. Glasgow Coma Score-measured neurological severity is the main determinant of future functional capacity, which is greater at 1 year. PMID- 12756438 TI - Comment on "Prevention of severe Candida infections in non-neutropenic, high risk, critically ill patients," by Garbino et al. PMID- 12756439 TI - Cardiac massage in infants. PMID- 12756440 TI - Effects of helium-oxygen on respiratory mechanics, gas exchange, and ventilation perfusion relationships in a porcine model of stable methacholine-induced bronchospasm. AB - OBJECTIVE: To explore the consequences of helium/oxygen (He/O(2)) inhalation on respiratory mechanics, gas exchange, and ventilation-perfusion (VA/Q) relationships in an animal model of severe induced bronchospasm during mechanical ventilation. DESIGN: Prospective, interventional study. SETTING: Experimental animal laboratory, university hospital. INTERVENTIONS: Seven piglets were anesthetized, paralyzed, and mechanically ventilated, with all ventilator settings remaining constant throughout the protocol. Acute stable bronchospasm was obtained through continuous aerosolization of methacholine. Once steady-state was achieved, the animals successively breathed air/O(2) and He/O(2) (FIO(2) 0.3), or inversely, in random order. Measurements were taken at baseline, during bronchospasm, and after 30 min of He/O(2) inhalation. RESULTS: Bronchospasm increased lung peak inspiratory pressure (49+/-6.9 vs 18+/-1 cm H(2)O, P<0.001), lung resistance (22.7+/-1.5 vs 6.8+/-1.5 cm H(2)O x l(-1).s, P<0.001), dynamic elastance (76+/-11.2 vs 22.8+/-4.1 cm H(2)O x l(-1), P<0.001), and work of breathing (1.51+/-0.26 vs 0.47+/-0.08, P<0.001). Arterial pH decreased (7.47+/ 0.06 vs 7.32+/-0.06, P<0.001), PaCO(2) increased, and PaO(2) decreased. Multiple inert gas elimination showed an absence of shunt, substantial increases in perfusion to low VA/Q regions, and dispersion of VA/Q distribution. He/O(2) reduced lung resistance and work of breathing, and worsened hypercapnia and respiratory acidosis. CONCLUSIONS: In this model, while He/O(2) improved respiratory mechanics and reduced work of breathing, hypercapnia and respiratory acidosis increased. Close attention should be paid to monitoring arterial blood gases when He/O(2) is used in mechanically ventilated acute severe asthma. PMID- 12756441 TI - Candiduria in critically ill patients admitted to intensive care medical units. AB - BACKGROUND: The purpose of this study was to determine the incidence of candiduria in critically ill patients admitted to intensive care medical units (ICUs), to identify risk factors for candiduria and to assess the frequency distribution of different Candidaspp. SUBJECTS AND METHODS: This was a prospective cohort observational and multicenter study. A total of 1,765 patients older than 18 years of age who were admitted for at least 7 days to 73 medical surgical ICUs of 70 Spanish hospitals were included in the study. Urine cultures were performed once a week. RESULTS: In 389 patients (22%), Candidaspp. in one or more urine samples were isolated. In the multivariate analysis, independent risk factors for candiduria included: age >65 years, female sex, length of hospital stay before ICU admission, diabetes mellitus, total parenteral nutrition, mechanical ventilation and previous use of antimicrobials. Candida albicanswas recovered in 266 cases (68.4%), followed by C. glabrata(32 cases, 8.2%) and C. tropicalis(14 cases, 36%). Previous use of antifungal agents was the only risk factor for the selection of Candidanon-albicans candiduria (OR 2.64, 95% CI 1.35 5.14, P=0.004). In-hospital mortality was 48.8% in patients with candiduria compared to 36.6% in those without candiduria ( P<0.001). Significant differences were also found for ICU mortality (38.% vs. 28.1%, P<0.001). CONCLUSIONS: Twenty two percent of critically ill patients admitted for more than 7 days in the ICU developed candiduria. C. albicanswas the most frequent causative pathogen. Previous use of antifungals was the only risk factor for the selection of Candidanon-albicans. PMID- 12756442 TI - Vibrio harveyi mutagenicity assay as a preliminary test for detection of mutagenic pollution of marine water. PMID- 12756444 TI - Comparative quantitative analysis of agricultural chemicals using a microplate mammalian cell cytotoxicity assay. PMID- 12756443 TI - Nuclear anomalies and blood protein variations in fish of the Hooghly-Matlah river system, India, as an indicator of genotoxicity in water. PMID- 12756445 TI - Relative changes of elements in human osseous tissue. PMID- 12756447 TI - Adsorptive behavior of acetochlor on organoclay complexes. PMID- 12756446 TI - Polychlorinated biphenyls in taiwanese primipara human milk and associated factors. PMID- 12756448 TI - Decolorization of acid blue 9 dye wastewater using waste furnace slag. PMID- 12756449 TI - Improved analytical method for residual dioxins in human milk. PMID- 12756450 TI - Photodegradation of diesel oil in aqueous solutions. PMID- 12756451 TI - Evaluation of beta-cyfluthrin: protection of cole crops, dietary intake, and consumer risk assessment. PMID- 12756452 TI - Maximum residue limit and risk assessment of beta-cyfluthrin and imidacloprid on tomato (Lycopersicon esculentum Mill). PMID- 12756453 TI - Polychlorinated biphenyls and organochlorine pesticides in edible fish species and dolphins from Guanabara Bay, Rio de Janeiro, Brazil. PMID- 12756454 TI - Persistent organochlorine pollutants in the aquatic ecosystem of Lake Manzala, Egypt. PMID- 12756455 TI - Airborne exposure concentrations of lead, cadmium, and chromium during demolition of incinerators inside a building. PMID- 12756456 TI - Asbestos contamination in biota and abiota in the vicinity of asbestos-cement factory. PMID- 12756457 TI - Influence of leaded-gasoline regulations on the blood lead concentrations in Murciano-Granadina goats from Murcia Region, Southeast Spain. PMID- 12756458 TI - Effect of pan masala on sperm morphology of a mouse. PMID- 12756459 TI - Protective role of ascorbic acid against lead toxicity in blood of albino mice as revealed by metal uptake, lipid profiles, and ultrastructural features of erythrocytes. PMID- 12756461 TI - In vitro effect of azadirachtin on aerobic bacteria of rat intestine. PMID- 12756460 TI - Biodisposition and biochemical effects of a new phosphoramidate series in rat tissues. PMID- 12756462 TI - Mercury in bottom sediments of the tropical Rio Marabasco, its estuary, and Laguna de Navidad, Mexico. PMID- 12756463 TI - Heavy metal distribution in surface sediments from Mtwapa and Shirazi Creeks, Kenyan coast. PMID- 12756464 TI - Trace metals in sediments from Bahia de Chetumal, Mexico. PMID- 12756465 TI - Total metals in intertidal surface sediment of oyster culture areas in Sonora, Mexico. PMID- 12756466 TI - Predicting mercury in mallard ducklings from mercury in chorioallantoic membranes. PMID- 12756467 TI - Sensitivity of Ceriodaphnia dubia of different ages to sodium chloride. PMID- 12756468 TI - Structure-toxicity relationships for the effects of N- and N,N'-alkyl thioureas to Tetrahymena pyriformis. PMID- 12756469 TI - Acute Toxicity of Carbofuran to a Freshwater Teleost, Clarias batrachus. PMID- 12756470 TI - Acute and chronic toxicity of dimethylsulfoxide to Daphnia magna. PMID- 12756471 TI - Development of high-oleic, low-linolenic acid Ethiopian-mustard (Brassica carinata) germplasm. AB - Seed oil of current zero erucic-acid germplasm of Ethiopian mustard ( Brassica carinata A. Braun) is characterized by a low concentration of oleic acid and high concentrations of linoleic and linolenic acids. Sources of increased oleic-acid (HO) and reduced linolenic-acid (LL) concentration have been developed separately in high erucic-acid germplasm. The objectives of the present research were to study the inheritance of the HO and LL traits in crosses HO x LL, and to develop HOLL recombinants, both in high erucic-acid and zero erucic-acid backgrounds. The HO mutant N2-3591 (about 20% oleic acid compared to 9% in conventional high erucic-acid materials), was reciprocally crossed with the LL lines N2-4961 and HF 186 (both with about 5% linolenic acid compared to 12% in standard high erucic acid materials). Increased oleic acid concentration of N2-3591 was found to be controlled by alleles at one locus (Ol), whereas three different loci for reduced linolenic-acid concentration (Ln, Ln1 and Ln2) were identified in N2-4961 and HF 186. Crosses between N2-3591 and N2-4961 generated HOLL recombinants where levels of increased oleic-acid and reduced linolenic-acid were similar to those of the parents. However, a transgressive segregation for oleic acid was observed in crosses between N2-3591 and HF-186, where F(2) seeds with up to 29.7% oleic acid were obtained, in comparison to an upper limit of 25.1% in the N2-3591 parent grown in the same environment. The transgressive increased oleic-acid was expressed in the F(3) generation and was attributed to the presence of a second locus, designated Ol2. The transgressive trait was transferred to the zero erucic acid line 25X-1, resulting in a zero erucic-acid germplasm with very high oleic acid concentration (83.9% compared to 32.9% in 25X-1) and low linolenic-acid concentration (5.0% compared to 16% in 25X-1). Additionally, two other lines exhibiting different stable levels of increased oleic-acid (70.7% and 79.5%, respectively) and reduced levels of linolenic-acid (7.5% and 8.7%, respectively) were isolated. PMID- 12756472 TI - Biochemical and genetic studies of two Heterodera avenae resistance genes transferred from Aegilops ventricosa to wheat. AB - Two Heterodera avenae resistance genes, Cre2 from Aegilops ventricosa AP-1 and Cre5 from Ae. ventricosa #10, were shown to confer a high level of resistance to the Spanish pathotype Ha71. No susceptible plants were found in the F(2) progeny from the cross between the two accessions of Ae. ventricosa, suggesting that their respective resistance factors were allelic. However, genes Cre2 and Cre5 apparently were transferred to a different chromosomal location in the wheat line H-93-8 and in the 6M(v)(6D) substitution, respectively, as proved by F(2) segregation of their cross progeny. The induction of several defence responses during early infection by the same H. avenae pathotype in resistant lines carrying Cre2 or Cre5 genes was studied. Isoelectrofocusing (IEF) isozyme analysis revealed that peroxidase, esterase and superoxide dismutase activity increased after nematode infection, in roots of resistant lines in comparison with their susceptible parents. Differential induced isoforms were also identified when IEF patterns of resistant lines were compared. A DNA marker, absent in Cre5-carrying genotypes, was found to be linked, thought not very tightly, to the Cre2 gene in the H-93-8 line. The differences observed between the Cre2 and Cre5 genes with respect to their chromosomal location in wheat introgression lines, de-toxificant enzyme induction and behaviour against different pathotypes, suggest they are different H. avenae resistance sources for wheat breeding. PMID- 12756474 TI - [Cardiovascular changes during pregnancy]. AB - Cardiovascular alterations during pregnancy are characterized by an increased vascular volume, cardiac output, and heart rate, with a marked fall in vascular resistance. Cardiac output is about 40-50% higher during the third trimester. Even higher values of cardiac output are observed during uterine contractions in labor. In general, arterial blood pressure remains unaffected or demonstrates some tendency toward lower diastolic pressure. The higher blood volume is associated with a slight increase in left ventricular dimensions. Left ventricular contraction force and its first derivative remain unchanged. Many symptoms and findings during pregnancy are caused by the described changes, such as dyspnea on exertion, presyncope due to pressure on the inferior vena cava resulting in a decreased venous return to the heart, prominent jugular venous pulsation, leg edema, and ejection murmurs over the aorta and pulmonary artery. Paroxysmal nocturnal dyspnea, anginal chest pain, syncopy, anasarca, and diastolic heart murmurs require further evaluation. PMID- 12756473 TI - Uncoupling protein 1 and 3 polymorphisms are associated with waist-to-hip ratio. AB - Body weight regulation is a complex phenotype also depending on the action of uncoupling proteins (UCPs) that mediate the "uncoupling" of respiration leading to the dissipation of energy as heat. This study investigated whether genetic variants in the genes encoding UCP-1 and UCP-3 are associated with different obesity-related phenotypes in 162 whites with a wide range of body mass index. All subjects were genotyped for the polymorphisms UCP-1 A-3826G, UCP-1 Ala64Thr, and UCP-3 C-55T using a PCR-based restriction method with appropriate enzymes. The frequencies of the UCP-1 3826G, UCP-1 64Thr, and UCP-3 55T alleles were 27.2%, 12.0%, and 22.8%, respectively. No significant associations were observed between polymorphism and body mass index or obesity. However, after adjustment for gender, age, body mass index, and diabetes mellitus the waist-to-hip ratio was significantly associated with UCP-1 Ala64Thr ( P=0.003) and UCP-3 C-55T ( P=0.02) but not with UCP-1 A-3826G. The higher waist-to-hip ratios associated with the UCP-1 64Thr and UCP-3 55T alleles were due to higher waist circumference in these allele carriers. In conclusion, central obesity in whites as reflected by an increased waist-to-hip ratio is associated with the UCP-1 Ala64Thr and UCP 3 C-55T polymorphisms. To what extent these genotypes contribute to the overall cardiovascular risk remains to be elucidated. PMID- 12756475 TI - [Myocardial infarction and thromboembolism during pregnancy]. AB - Acute myocardial infarction is a very rare event during pregnancy and bears the problem of misdiagnosis. However, about 150 cases have been published worldwide with a preponderance of anterior wall infarcts. With more women delaying childbearing until an older age and increasing prevalence of smoking in young women, it can be expected that all forms of coronary artery disease--including acute myocardial infarction--will be seen more often in the future. Among the causes of coronary artery occlusion in pregnancy are (1) rupture of very small coronary artery plaques triggered by different events, e.g., hypertension; (2) plain coronary artery disease; (3) dissection of coronary arteries; (4) coronary artery spasms with/without arterial thrombosis. Prompt diagnosis and immediate therapy are necessary to lower the high mortality of mother and fetus. The gold standard in the therapy of acute myocardial infarction during pregnancy is immediate coronary angiography and percutaneous transluminal coronary angioplasty (PTCA) with or without stent implantation. Application of thrombolytics (recombinant tissue plasminogen activator [rt-PA], r-PA, streptokinase [SK], urokinase [UK]) has been reported in single patients but should be limited to cases where acute PTCA is not available and where the infarct occurs before the 14th week of pregnancy because of possible embryopathy. If the patient is in the last 10 weeks of pregnancy, anticipation of delivery should be part of the medical planning. Consultation with an obstetrician must be obtained as soon as the patient enters the hospital. Besides bleeding complications, venous thrombosis with pulmonary embolism is among the most common causes of death during pregnancy. Pregnancy-related changes in physiology - increase in the resistance to flow from the lower extremities to the heart - and congenital coagulation abnormalities are most important to be recognized. This leads to the fact that superficial and deep venous thromboses occur more often in pregnancy than in the nonpregnant state. Among the coagulation abnormalities found in pregnancy are hypercoagulability (increased levels of fibrinogen, factor VII, factor VIII, factor X), decreased fibrinolytic activity due to an increased level of plasminogen activator inhibitor, increased adhesion and aggregation of platelets, decreased level of protein C and of the APC (activated protein C) ratio. Individual risks factors justifying diagnostic screening include contraception, smoking, immobilization, infection, adiposity, placental insufficiency, and a family history of thrombosis. It is even more important to establish/rule out the diagnosis of thrombosis in pregnancy than in the nonpregnant state, because the use of anticoagulants carries certain risks during pregnancy. Doppler vein studies should be used for diagnosis. If necessary, venography may be used with shielding of the maternal abdomen. Therapy consists of subcutaneous application of heparin, compression, and early mobilization. Alternatively, especially for long-term management, treatment with low molecular weight heparins is feasible. Thrombolytic treatment is contraindicated in most cases due to the high risk of bleeding complications. However, the application of thrombolytics can be contemplated in single cases after careful consideration of the pros and cons. Most cases of pulmonary embolism should also be handled conservatively with heparin. Only in massive pulmonary embolism with severe hemodynamic compromise, thrombolytic treatment is indicated. To guide future therapy in the patients, it is necessary to establish the lifetime risk of recurrent events by determining: APC resistance, prothrombin mutation 20210 A, homocysteine, AT III, protein C and S, antiphospholipid antibodies, and anticardiolipin antibodies. PMID- 12756476 TI - [Hypertension and hypertensive emergencies in pregnancy]. AB - BACKGROUND: Hypertensive emergencies are acute, life threatening, and usually- but not necessarily--associated with severe increases in blood pressure. In pregnancy, this is the fact in eclampsia. Eclampsia refers to the occurrence of one or more generalized convulsions in the setting of preeclampsia with proteinuria, edema, and hypertension. PATHOGENESIS: Our current understanding of the pathogenesis of preeclampsia will be reviewed here. Some major risk factors for the development are preexisting hypertension and renal disease. PREVENTION AND THERAPY: Preventive measures of preeclampsia and treatment of this specific hypertensive emergency in pregnancy are discussed. PMID- 12756477 TI - [Pregnancy and cardiomyopathies]. AB - This overview on the topic of cardiomyopathy and gestation comprises the diagnostic and therapeutic options of patients with preexistent cardiomyopathies (dilated, hypertrophic, inflammatory, and others) and with cardiomyopathies which have been discovered during or in the 6 months following delivery. CARDIOMYOPATHIES PREEXISTENT BEFORE GESTATION: If cardiomyopathy is present before an intended gestation, the couple should be advised against pregnancy because of the high risk of deterioration both during gestation and peripartum. If pregnancy occurs, according to ESC (European Society of Cardiology) recommendations termination should be advised if the ejection fraction is < 50% and/or the LV dimensions are definitely above normal. If termination is refused, the patient must be checked regularly by both gynecologist and cardiologist, by the latter to perform regular echocardiograms. Termination is not recommended for the hypertrophic (nonobstructive) cardiomyopathies. If atrial fibrillation occurs, anticoagulation with low molecular weight heparin and digoxin and/or Betablockers are recommended for rhythm and rate control. PERIPARTUM CARDIOMYOPATHIES: In peripartum cardiomyopathies, which are discovered clinically postpartum, inflammation of the myocardium sometimes associated with pericarditis is frequently found. For those patients, we recommend heart catheterization with endomyocardial biopsy to allow for the exact diagnosis of the underlying cardiac process (inflammatory and/or viral vs autoreactive myocarditis or noninflammatory or nonviral [= idiopathic] forms). This diagnostic algorithm, which we recommend for any form of dilated cardiomyopathy, bears impact on treatment options beyond the mere heart failure therapy that should be instigated anyhow. PMID- 12756478 TI - Pericardial disease in pregnancy. AB - BACKGROUND: There is no evidence that pregnancy affects susceptibility to pericardial disease. However, when such a condition occurs, its proper diagnosis and management may be crucial for the outcome of the pregnancy. INCIDENCE AND DIAGNOSIS: Hydropericardium is the most frequent form of pericardial involvement in pregnancy. It is typically a small, clinically silent pericardial effusion present in the third trimester in approximately 40% of healthy pregnant women. Small amounts of fetal pericardial fluid (< 2 mm in echocardiography, in diastole) can be detected after 20 weeks of gestation. Larger effusions should raise clinical concern for hydrops fetalis, Rh disease, hypoalbuminemia, and infectious or autoimmune disorder. Wide varieties of etiologic forms of pericardial diseases occur sporadically in pregnant women. Significant symptoms, electrocardiographic changes, or physiologic impairment warrant hospitalization. TREATMENT: Most pericardial disorders are managed during pregnancy as in nonpregnant patients (i.e., nonsteroidal antiinflammatory drugs for acute, antibiotics and drainage for purulent pericarditis, and corticosteroids for systemic autoimmune disorders). However, colchicine is contraindicated in pregnancy, and pericardiocentesis should be performed only for very large effusions causing clinical signs of cardiac tamponade or if presence of suppurative, tuberculous or neoplastic pericardial effusion is suspected. Echocardiographic guidance of pericardiocentesis is preferred to fluoroscopic guidance in order to avoid fetal X-ray exposure. Pericardiectomy should be reserved for significant pericardial constriction and resistant bacterial infections. Delivery of normal infants in term after pericardiocentesis or pericardiectomy is expected, whenever natural history of causative disease allows. Pericardiectomy itself is not a contraindication for subsequent successful pregnancies. PMID- 12756479 TI - [Cardiac arrhythmias during pregnancy--what to do?]. AB - METHODS: Atrial premature beats are frequently diagnosed during pregnancy, supraventricular tachycardia (atrial tachycardia, AV nodal reentrant tachycardia, circus movement tachycardia) less frequently. For acute therapy, electrical cardioversion with 50-100 J is indicated in all unstable patients. In stable supraventricular tachycardia, initial therapy includes vagal maneuvers to terminate breakthrough tachycardias. For short-term management, when vagal maneuvers fail, intravenous adenosine is the drug of first choice and may safely terminate the arrhythmia. For long-term therapy, beta-blocking agents with beta(1) selectivity are first-line drugs; class Ic agents or the class III drug sotalol represent effective and therapeutic alternatives. Ventricular premature beats are also frequently present during pregnancy and benign in most of the unstable patients; however, malignant ventricular tachyarrhythmias (sustained ventricular tachycardia, ventricular flutter, ventricular fibrillation) are less frequently observed. Electrical cardioversion is necessary in all patients with hemodynamically unstable situation and life-threatening ventricular tachyarrhythmias; in hemodynamically stable patients, initial therapy with ajmaline, procainamide or lidocaine is indicated. If prophylactic therapy is needed, beta-blocking agents with beta(1) selectivity are regarded as drugs of first choice. If this therapy proves ineffective, class Ic agents or sotalol can be considered. In patients with syncopal ventricular tachycardia, ventricular fibrillation, ventricular flutter or aborted sudden death, an implantable cardioverter-defibrillator is indicated. In patients with symptomatic bradycardia, a pacemaker can be implanted using echocardiography at any stage of pregnancy. CONCLUSIONS: The treatment of the pregnant patient with cardiac arrhythmias requires important modifications of the standard practice of arrhythmia management. The goal of therapy is to protect the patient and fetus through delivery, after which chronic or definitive therapy can be administered. PMID- 12756480 TI - [Congenital heart disease and acquired valvular lesions in pregnancy]. AB - BACKGROUND: In Germany, about 6,000 pregnancies in women with grown-up congenital heart disease or acquired valvular lesions are expected per year. The pregnancy related physiology is characterized by a 50% increase in plasma volume and a 25% increase in erythrocyte volume. The cardiac output increases by 40% due to 30% increase in stroke volume and 10% increase in heart rate during the first half, and 10% increase in stroke volume but 30% increase in heart rate during the second half of pregnancy. As a consequence of the decrease of systemic vascular resistance, the systolic and, even more, the diastolic blood pressures are reduced during approximately the first 20 weeks of pregnancy. UNCORRECTED CONGENITAL LESIONS: Women with uncorrectable congenital heart disease, congestive heart failure (NYHA III and IV) despite optimized medical treatment after palliative surgery, or pulmonary vascular resistances > 800 dyn x s x cm(-5) should be advised against pregnancy. The presence of congestive heart failure or persistent cyanosis in the mother are the most important predictors of fetal hypoxia. Patients with pretricuspid shunts (e.g., atrial septal defect [ASD]) are at low risk of a hemodynamic deterioration or first onset of arrhythmias. In the rare case of a marked clinical deterioration, catheter-based closing of the shunt is the first-line treatment. Also, ventricular septal defects and persistent ducti arteriosi are usually well tolerated during pregnancy, as they are highly resistant to flow. In some cases, arrhythmias may occur. The prognosis is less favorable, if myocardial compromise has already been present before pregnancy. The fatal complication rate correlates closely with the degree of congestive heart failure. In aortic coarctation, development of severe hypertension, myocardial decompensation, aortic dissection, and cerebral hemorrhage have been reported in 2.3% of cases. To prevent aortic dissection and rupture of cerebral vascular aneurysms, patients should be advised to reduce their physical activity and have their blood pressure controlled closely. If, during pregnancy, a therapeutic intervention is unavoidable, stent placement is the therapy of choice. The maternal complication rate is low in pulmonary artery stenosis. Hemodynamically significant stenoses should be treated before pregnancy. In the rare case of progressive right heart failure or cyanosis during pregnancy, balloon valvotomy is the first-line therapeutic option. CONGENITAL HEART DISEASE WITH PRIOR PALLIATION: Women with incomplete correction of a tetralogy of Fallot or significant residual gradients or shunts carry a particular risk of myocardial deterioration. A maternal hematocrit > 60%, an arterial O(2) saturation < 80%, markedly elevated right ventricular pressures, and the former presentation of syncopes are indicators of a poor prognosis. Fatal complication rates have been reported in 3-17% of cases. Other cyanotic lesions have been linked with a poor maternal and fetal prognosis. A 32% incidence of severe cardiovascular complications (pump failure, thromboembolic events, life-threatening arrhythmias, infective endocarditis) has been reported during 96 pregnancies of women with cyanotic heart disease. In addition, the frequency of abortions, premature birth, fetal distress, and congenital malformation of the child was 57%. ACQUIRED VALVE LESIONS: Mitral stenosis is the lesion that most frequently requires therapeutic intervention during pregnancy, as the transmitral flow increases and time of diastole decreases during pregnancy due to the increase in cardiac output and heart rate. A consequent increase in mean pulmonary artery pressure by approximately 50% and a deterioration by one to two NYHA classes must be expected. While patients with a mitral orifice area > 1.5 cm(2) can usually be treated medically, more advanced mitral stenoses often require percutaneous mitral balloon valvotomy, a procedure with a very low complication rate in experienced centers. A chronic mitral or aortic regurgitation without jeopardized myocardial function is usually well tolerated during pregnancy, as the drop in peripheral vascular resistance results in a favorable left ventricular impedance, which reduces the transmitral regurgitant fraction and improves left ventricular antegrade ejection. Moreover, the increase in heart rate limits diastolic transaortic regurgitation. Hemodynamically advanced aortic stenosis is rare among patients in child-bearing age. The hemodynamic changes during pregnancy result in a decrease of the transaortic flow per time and thus in a decrease of the transaortic pressure loss. On the other hand, myocardial wall stress and oxygen consumption are significantly increased. If aortic valve orifice area is > 1.5 cm(2), the hemodynamic situation is usually well tolerated during pregnancy. In the case of more advanced aortic stenosis, there is a considerable risk of myocardial decompensation. The development of symptoms such as dyspnea, near syncopes or syncopes, and arrhythmias are indicators of a complicated course. If treatment is unavoidable, aortic valve replacement is the therapy of choice. ORAL ANTICOAGULATION: With respect to anticoagulation during pregnancy, there is an ongoing debate about the potential risk and benefit of phenprocoumon, standard heparins, and low molecular heparins. Withdrawal of any anticoagulation results in the most favorable fetal outcome, oral anticoagulation throughout pregnancy in the best prognosis for the mother. An individual approach by an experienced center taking all therapeutic options into account is probably the best strategy. PMID- 12756481 TI - [Two- and three-dimensional echocardiographic analysis of congenital heart disease in the fetus]. AB - INTRODUCTION: With increasing experience of obstetric sonographers, a higher proportion of cardiac malformations is found antenatally. However, undiagnosed fetal cardiac defects still result in a significant pre- and postnatal morbidity and mortality. The purpose of two- and three-dimensional echocardiographic imaging in the fetus is to provide clear representations of the underlying cardiac and vascular anatomy. Studies on pre- and postnatal echocardiography have shown these techniques to provide an adequate form of image display for comprehensive assessment of most cases with congenital heart disease. METHODS: To date, two different methods are used for three-dimensional echocardiography in the fetus. The technique currently employed at numerous institutions derives from a complex assembly of sequentially acquired and reconstructed two-dimensional images and is analogous to the 3-D technology assessed in studies on neonates, children, and adults. Although an electromagnetic location device is used to register transducer position during data acquisition, this technique has important limitations due to fetal movement artifacts and difficulties in cardiac gating. This often results in inadequate image quality when compared with 2-D echocardiography. Recent progress in the design and fabrication of higher frequency real-time volumetric transducers has greatly improved 3-D echocardiographic imaging resolution and allows more immediate three-dimensional "on-line" analysis of cardiac anatomy. CONCLUSIONS: Advantages of 3-D fetal echocardiography include the ability to slice the acquired 3-D volume data into an infinite number of two-dimensional cross sections, and the ability to reconstruct unique three-dimensional views not seen with two-dimensional imaging. However, considering the current limitations and the time needed for 3-D image processing, its practical clinical relevance in the antenatal situation is not yet clear. PMID- 12756482 TI - Prenatal diagnosis of the fetus with hypoplastic left heart syndrome management and outcome. AB - OBJECTIVES: To review our 13-year experience with prenatally detected hypoplastic left heart syndrome (HLHS) of which management remains controversial. MATERIAL AND METHODS: Retrospective study of the management and outcome in all cases of HLHS diagnosed prenatally in a tertiary referral center for pediatric cardiology and cardiac surgery between January 1988 and July 2001. RESULTS: The diagnosis of HLHS was made in 32 fetuses. One mother had two pregnancies associated with HLHS. In 16 cases parents opted for termination of pregnancy and in five for compassionate care. Four fetuses died in utero, and seven patients received a palliative reconstructive Norwood procedure. In seven fetuses, associated anomalies were detected: three chromosomal and structural and four only structural. In six fetuses, other associated intracardiac anomalies were detected. Of seven infants operated, six had no associated anomalies and only one is alive at an age of 17 months. CONCLUSION: The low percentage of intention to treat among patients in our center (34%) is in accordance with the percentage found in another study from the UK (36.2%), but differs significantly from reported series across the Atlantic (67%). Prenatal diagnosis of the HLHS provides opportunities not only for getting patients in optimal preoperative condition when surgery is offered, but also for in-depth counseling of the parents on this severe malformation. A minority of parents faced with the difficult decision of possible termination of pregnancy, compassionate care or the Norwood strategy, choose surgical treatment which might be based on socioreligious differences and the interpretation of the long-term quality of life. PMID- 12756483 TI - [Anomalous origin of the left coronary artery. Surgical therapy using a miniaturized heart-lung machine]. AB - Anomalies of the coronary arteries are rare, congenital heart diseases which often require surgical therapy. We report on a patient with an aberrant left coronary artery originating from the right coronary artery and discuss the operative procedure using a miniaturized heart-lung machine. PMID- 12756484 TI - Activation of Na+/H+ exchange by protein phosphatase inhibitors in red blood cells of the frog Rana ridibunda. AB - The present study was designed to evaluate the role of protein phosphatases in regulation of sodium transport in the marsh frog erythrocytes using 22Na as a tracer. For this purpose the cells were treated with several known inhibitors of protein phosphatases. In standard isotonic medium, exposure of the cells to 10 mmol l(-1) NaF, 20 nmol l(-1) calyculin A or 0.1 mmol l(-1) cantharidin resulted in a significant (1.7-fold) increase in unidirectional ouabain-insensitive Na+ influx. The Na+ influx in frog red cells was progressively activated as the medium osmolality was increased by addition of 100, 200 or 300 mmol l(-1) sucrose to standard isotonic medium. The stimulatory effect of protein phosphatase blockers on Na+ influx was much higher in hypertonic medium containing 100 or 200 mmol l(-1) sucrose than that in isotonic medium. Stimulation of Na+ transport enhanced with increasing concentrations of calyculin A, and half-maximal activation (EC50) was obtained at 16 nmol l(-1). However, Na+ influx induced by strong hypertonic treatment (+300 mmol l(-1) sucrose) was not altered further in the presence of protein phosphatase inhibitors. The changes in Na+ influx evoked by protein phosphatase inhibitors and hypertonic treatment were associated with a rise in the intracellular Na+, but not K+, content. Enhancement in Na+ influx after addition of protein phosphatase blockers to cell suspension in isotonic or hypertonic media was almost completely inhibited by Na+/H+ exchange inhibitors, amiloride and ethyl-isopropyl-amiloride. The basal Na+ influx in frog erythrocytes in isotonic medium was relatively low (1.7 mmol/l cells/h) and not affected by 1 mmol l(-1) amiloride. Thus, the data obtained clearly indicate that Na+/H+ exchanger in the marsh frog red blood cells is under tight regulatory control, in all likelihood via protein phosphatases of types PP-1 and PP-2A. PMID- 12756485 TI - Effects of temperature acclimation on maximum heat production, thermal tolerance, and torpor in a marsupial. AB - Marsupials, unlike placental mammals, are believed to be unable to increase heat production and thermal performance after cold-acclimation. It has been suggested that this may be because marsupials lack functional brown fat, a thermogenic tissue, which proliferates during cold-acclimation in many placentals. However, arid zone marsupials have to cope with unpredictable, short-term and occasionally extreme changes in environmental conditions, and thus they would benefit from an appropriate physiological response. We therefore investigated whether a sequential two to four week acclimation in Sminthopsis macroura (body mass approx. 25 g) to both cold (16 degrees C) and warm (26 degrees C) ambient temperatures affects the thermal physiology of the species. Cold-acclimated S. macroura were able to significantly increase maximum heat production (by 27%) and could maintain a constant body temperature at significantly lower effective ambient temperatures (about 9 degrees C lower) than when warm-acclimated. Moreover, metabolic rates during torpor were increased following cold-acclimation in comparison to warm-acclimation. Our study shows that, despite the lack of functional brown fat, short-term acclimation can have significant effects on thermoenergetics of marsupials. It is likely that the rapid response in S. macroura reflects an adaptation to the unpredictability of the climate in their habitat. PMID- 12756486 TI - Design of the Jacky dragon visual display: signal and noise characteristics in a complex moving environment. AB - Visual systems are typically selective in their response to movement. This attribute facilitates the identification of functionally important motion events. Here we show that the complex push-up display produced by male Jacky dragons ( Amphibolurus muricatus) is likely to have been shaped by an interaction between typical signalling conditions and the sensory properties of receivers. We use novel techniques to define the structure of the signal and of a range of typical moving backgrounds in terms of direction, speed, acceleration and sweep area. Results allow us to estimate the relative conspicuousness of each motor pattern in the stereotyped sequence of which displays are composed. The introductory tail flick sweeps a large region of the visual field, is sustained for much longer than other components, and has velocity characteristics that ensure it will not be filtered in the same way as wind-blown vegetation. These findings are consistent with the idea that the tail-flick has an alerting function. Quantitative analyses of movement-based signals can hence provide insights into sensory processes, which should facilitate identification of the selective forces responsible for structure. Results will complement the detailed models now available to account for the design of static visual signals. PMID- 12756488 TI - Tomatoes, lycopene and prostate cancer: a clinician's guide for counseling those at risk for prostate cancer. AB - Prostate cancer has become a major public health issue and the search for etiologic risk factors and the development of chemopreventive agents has gained momentum over the last decade. An important epidemiologic finding has been the association between the consumption of tomato products and a lower risk of prostate cancer. Several investigators have proposed that lycopene, a carotenoid consumed largely from tomato products, may be the component responsible for lowering the risk of prostate cancer. Laboratory and clinical studies have been initiated with the goal of assessing the ability of pure lycopene to serve as a chemopreventive agent for prostate cancer. The focus on lycopene should continue, and an improved understanding of lycopene absorption, distribution, role in antioxidant reactions, and metabolism is critical in the quest to elucidate mechanisms whereby this compound may possibly reduce prostate cancer risk. PMID- 12756487 TI - Odor-guided behavior in Drosophila requires calreticulin. AB - The efficient processing of olfactory information is crucial for many aspects of life in animals, including behavior in insects. While much is known about the organization of the insect olfactory system, comparatively little is understood about the molecules that support its function. To further elucidate the molecular basis of olfaction, we explored the role of the calcium-binding chaperone calreticulin in the behavioral response of Drosophila to aversive odorants. We show that avoidance of naturally aversive odorants is impaired in flies harboring mutations in Calreticulin. Calreticulin mutants have broad defects in odor avoidance without abnormalities in antennal responses to odorants, alterations in central nervous system structure, or deficits in overall locomotor abilities. Interestingly, Calreticulin mutants exhibit defects in behavioral responses to odorants at low strength, whereas responses to higher odorant concentrations are preserved in these animals. Our studies indicate that calreticulin plays a key role in olfactory system function, possibly by establishing its overall sensitivity to odorants. PMID- 12756489 TI - High-grade prostatic intraepithelial neoplasia and prostate cancer risk reduction. AB - High-grade prostatic intraepithelial neoplasia (PIN) is the earliest accepted stage in carcinogenesis, possessing most of the phenotypic and biochemical changes in cancer without invasion of the basal membrane of the acini. The support for high-grade PIN as the main premalignant lesion of prostate cancer is based on several lines of evidence derived from prostate cancer animal models, epidemiological, morphological, genetic, and molecular studies. The incidence of high-grade PIN averages 9% (range 4-16%) in prostate biopsies, representing 115,000 new cases of high-grade PIN diagnosed each year in the United States. Performing saturation prostate biopsies to rule out any coexistent prostate cancer followed by every 3-6 month serial repeated prostate biopsies is currently the only way in which to manage patients found to have high-grade PIN. Medical therapy for high-grade PIN may easily become the mainstay treatment for high grade PIN. Treatment of high-grade PIN appears to be of clinical benefit notwithstanding the potential for prostate cancer risk reduction. These clinical benefits would reduce morbidity, enhance quality of life, delay surgery or radiation, and increase the interval for surveillance requiring invasive procedures. PMID- 12756491 TI - The Prostate Cancer Prevention Trial: design, status, and promise. AB - The Prostate Cancer Prevention Trial is the first phase III (randomized, placebo controlled, prospective) population-based study to determine whether the risk of prostate cancer can be reduced. Between 1994 and 1997, 18,882 men were randomized to either finasteride or placebo at 219 sites in the United States. The study design included annual digital rectal examination and PSA determinations with PSA performed centrally with reports provided to sites corrected for treatment arm (as finasteride lowers PSA level). Because of a number of potential biases that could confound disease ascertainment all patients were scheduled for an end-of study prostate biopsy. End-of-study prostate biopsies began in 2000, and the final biopsy is expected in 2004, with results of the study anticipated shortly thereafter. The high participant interest and extensive data collected for this study demonstrate the extraordinary opportunity for chemoprevention trials of prostate cancer in the United States PMID- 12756490 TI - The selenium and vitamin E cancer prevention trial. AB - BACKGROUND: Evidence suggests that both selenium and vitamin E reduce the risk of prostate cancer. The Selenium and Vitamin E Cancer Prevention Trial (SELECT) is a randomized, prospective, double-blind study designed to determine whether selenium and vitamin E alone and in combination can reduce the risk of prostate cancer among healthy men. MATERIALS AND METHODS: The preclinical and epidemiological evidence supporting a role for selenium and vitamin E as chemopreventive agents in prostate cancer are reviewed, and details of the trial design are presented. RESULTS. Preclinical, epidemiological, and phase III data from randomized, placebo-controlled clinical trials suggest that both selenium and vitamin E have potential efficacy in prostate cancer prevention. SELECT is a 2x2 factorial study with an accrual goal of 32,400 men with nonsuspicious DRE and serum PSA of 4 ng/ml or lower. CONCLUSIONS: SELECT is the second large-scale study of chemoprevention for prostate cancer. Enrollment began in 2001 with final results anticipated in 2013. PMID- 12756493 TI - The non-steroidal antiandrogen, bicalutamide ('Casodex'), may preserve bone mineral density as compared with castration: results of a preliminary study. AB - The impact of bicalutamide (Casodex) monotherapy on bone mineral density (BMD) was investigated in patients with locally advanced prostate cancer. BMD was assessed after treatment with bicalutamide 150 mg daily ( n=21) or by medical castration (goserelin acetate 3.6 mg every 28 days) ( n=8) for a median of 287 weeks. In 38% of castration compared with 17% of bicalutamide patients, femoral neck Z-scores were < or =-1 SD of the reference value (accepted as a two to three times increased risk of fracture) and T-scores were < or =-2.5 SD (World Health Organization definition of osteoporosis in white females). Total hip Z-scores were < or =-1 in 43% of castration patients and 13% of bicalutamide patients. In 38% of patients, lumbar spine BMD was affected by degenerative disease. These preliminary data suggest that there may be an advantage in terms of BMD in using bicalutamide monotherapy compared with castration; a benefit confirmed in a recent prospective randomised study. PMID- 12756494 TI - Increased discrimination between benign prostatic hyperplasia and prostate cancer with equimolar total prostate specific antigen measurement. AB - Although prostate specific antigen (PSA) is widely used in the discrimination of benign prostatic hyperplasia (BPH) and prostate cancer, its diagnostic value is controversial due to an appreciable false positive rate. In the present study, we compared a recently introduced assay method, equimolar PSA measurement, to non equimolar PSA measurement and also determined the diagnostic value of percent free PSA with changing total PSA (tPSA) measurements. Between April 1999 and December 2001, the sera of 61 patients with BPH and 41 with prostate cancer were examined. Total PSA and free PSA was determined using the Immulite 2000 assay system, whereas equimolar tPSA measurement was performed using Bayer PSA Q for the Chiron ACS 180 system. Comparative analysis of the two different assays revealed better diagnostic sensitivity and specificity values for equimolar tPSA measurement, which in turn would have led to 10% of the patients avoiding an unnecessary biopsy. Additionally, percent free PSA with the changing denominator of tPSA assays showed that the free PSA/equimolar tPSA ratio was the best tumor marker among the studied forms of PSA. It was concluded that equimolar tPSA measurement using recombinant Fab fragments is superior to the classical measurements with monoclonal antibodies, and that the use of percent free PSA with the equimolarly measured tPSA has better sensitivity and specificity in the discrimination of benign and malignant diseases of the prostate. PMID- 12756492 TI - Antiestrogens and selective estrogen receptor modulators reduce prostate cancer risk. AB - The development of chemoprevention strategies against prostate cancer would have the greatest overall impact both medically and economically against prostate cancer. Estrogens are required for prostate carcinogenesis. Estrogenic stimulation through estrogen receptor alpha in a milieu of decreasing androgens contributes significantly to the genesis of benign prostatic hyperplasia, prostate dysplasia, and prostate cancer. The ability of antiestrogens and selective estrogen receptor modulators (SERMs) to delay and to suppress prostate carcinogenesis is supported by preclinical, clinical, and epidemiological studies. SERMs have many features that make them attractive candidates for prostate cancer chemoprevention including their favorable safety profile and efficacy in preclinical prostate cancer models. The true clinical benefits of SERMs for chemoprevention to prevent prostate cancer, however, should continue to be investigated through human clinical trials. A phase IIb/III human clinical trial is currently evaluating safety and efficacy of toremifene, a SERM, in men who have high-grade prostatic intraepithelial neoplasia. PMID- 12756495 TI - Fibromyalgia in diabetes mellitus. AB - OBJECTIVE: The aim of this study was to evaluate the prevalence of fibromyalgia (FM) in patients with diabetes mellitus (DM). SUBJECTS: The study included 100 consecutive unselected patients with DM attending our diabetes clinic. Patients were divided into two groups: 45 patients with type 1 diabetes and 55 patients with type 2 diabetes. A group of 50 healthy hospital staff members served as controls. The FM was diagnosed according to the 1990 American College of Rheumatology criteria. Counts of 18 tender points were performed by thumb palpation and assessed by dolorimeter. Routine biochemical tests and levels of HbA(1c) were recorded in each patient. RESULTS: The main outcome measure was the association of FM with DM. Fibromyalgia was diagnosed in 17 patients (17%) with DM and in only one (2%) healthy control ( P=0.008). No differences in patients were noted in the prevalence of FM between type 1 and type 2 diabetes (18.5% vs 15.5%, respectively). Patients with both FM and DM had significantly higher levels of HbA(1c) than DM patients without FM (9.2+/-1.1% vs 6.4+/-1.5%) ( P<0.05). Similarly, the numbers of tender points, pain scores, and the prevalence of sleep disturbances, fatigue, and headaches were higher in this group of patients. A significant correlation was observed between the numbers of tender points and HbA(1c) levels in the DM patients with FM ( r=0.72, P=0.027). CONCLUSION: Fibromyalgia is a common finding in patients with types 1 and 2 diabetes, and its prevalence could be related to control of the disease. As with other diabetes complications, FM might be prevented by improved control of blood glucose levels. PMID- 12756496 TI - Comparison of different transformation methods for Aspergillus giganteus. AB - Four different transformation methods were tested and compared in an attempt to facilitate the genetic transformation of Aspergillus giganteus, the producer of an antifungal protein (AFP). The fungus was transformed to hygromycin B resistance, using the hph gene of Escherichia coli by protoplast transformation, electroporation, biolistic transformation, and Agrobacterium tumefaciens-mediated transformation. Electroporation and biolistic transformation were found to be inappropriate for transforming A. giganteus, due to a low transformation yield. The conventional transformation technique based on protoplasts yielded up to 55 transformants in 10(8) protoplasts/microg DNA and was enhanced to 140-fold by A. tumefaciens-mediated transfer of its T-DNA. Here, the germination time prior to cocultivation and the fungus:bacterium ratio were found to alter the transformation efficiency. Southern blot analysis revealed that the A. giganteus transformants contained a randomly integrated single T-DNA copy, whereas multiple integration events were frequent in transformants obtained by the protoplast method. PMID- 12756497 TI - Prognosis of older patients with acute myeloid leukemia receiving either induction or noncurative treatment: a single-center retrospective study. AB - Since few studies focus on prognostic factors in unselected elderly acute myeloid leukemia (AML) patients, a retrospective analysis of 138 consecutive patients aged >55 years (median age: 67, range: 56-89) with AML diagnosed at a single center over an 8-year period was performed: 69% had de novo AML and 31% secondary (s) AML; 67% of the patients were karyotyped. Of the patients, 73 (53%) were treated with standard induction therapy protocols and 65 (47%) received palliative treatment only. Univariate and multivariate analyses of the effects of the following factors on overall survival (OS) were performed: sex, age > or = vs <65 years, de novo vs sAML, serum (s) lactate dehydrogenase (LDH) > or = vs <400 U/l, leukocytes > or = vs <50,000/ microl, induction therapy, and karyotype. Additionally, in patients receiving induction therapy, complete remission (CR) rates and survival from CR were analyzed. CR rate was 47% [95% confidence interval (35%, 59%)], 53% (39%, 66%) in de novo AML, and 21% (5%, 51%) in sAML. After a median follow-up of 4 years, 130 deaths were observed (94%). In a univariate analysis, significant factors for longer OS were induction therapy, age <65 years, sLDH <400 U/l, and de novo AML. In a multivariate analysis, significant factors for longer OS were sLDH <400 U/l and induction therapy. However, the difference between treatment outcome may also be due to selection criteria not captured, such as performance status, comorbid conditions, wish of the patient, etc. The effects of intensive and nonintensive treatment in this patient group need to be investigated in prospective, randomized trials in which these clinical parameters of high relevance for treatment decisions in older patients are also considered. PMID- 12756499 TI - HOVON 47/EORTC 20013: chlorambucil vs 2x2 Gy involved field radiotherapy in stage III/IV previously untreated follicular lymphoma patients. PMID- 12756500 TI - Measurement of three-joint-finger motions: reality or fancy? A three-dimensional anatomical approach. AB - Various anatomical publications have reported two-dimensional studies with flexion/extension or abduction/adduction motion analysis, but longitudinal axial rotations (LAR) of three-joint fingers have rarely been mentioned. The aim of our study was to determine the maximal passive motions of three-joint-fingers and to measure the passive LAR of phalanges during a flexion/extension movement. A protocol of anatomical dissection was carried out with 22 fresh-frozen limbs from 11 human cadavers free from any visible pathology. The sample consisted of six females and five males with a mean age of 75.7 years (range 65-94 years). Passive motions of fingers excluding the thumb were analyzed with a wire circling technique. Extreme flexion/extension angles and adduction/abduction laxities were measured for each joint. LAR angles of distal bony segment position were evaluated in comparison with the proximal bony segment position in extreme flexion or extension. The results were recorded for the joints of each three joint-finger. No difference was statistically related to sex or right/left-sided criteria ( p>0.05). Passive LARs were measured in spite of an aggressive anatomical protocol. A small database was set up. LARs were an important third type of motion. They should be analyzed during a routine clinical examination of patients' hands as well as flexion/extension or abduction/adduction motions. PMID- 12756501 TI - Embryology of the walls of the lateral sellar compartment: apropos of a continuous series of 39 embryos and fetuses representing the first six months of intra-uterine life. AB - The aim of this study was a continuous and rather exhaustive description of the embryological development of the lateral sellar compartment. The histological sections of 39 embryos and fetuses were studied, and represent the first six months of intra-uterine life. The embryological period showed the organization of the content of the compartment. The medial and lateral walls appeared during the 15th week of amenorrhoea, and did not modify later. The medial wall was constituted in its rostral part by the hypophyseal lodge and in its caudal part by the periosteum of the sphenoid bone. Two layers formed the lateral wall: the superficial layer was an expansion of the dura mater that surrounded the oculomotor nerves along their course to the superior orbital fissure; the deep layer was weaker, and surrounded and joined the nerves together. The results of this study advocate the evolution of the nomenclature of this region, as the term "inter-periosto-dural space" would better reflect the real pattern of the lateral sellar compartment. Furthermore, the presence of a communication between the two lateral sellar compartments has led to a discussion of the previous hypothesis about the development of the lateral sellar compartment. The venous network was located on both the medial and lateral sides of the internal carotid artery, but expanded in the lateral wall of the lateral sellar compartment. That is of interest to surgeons and radiologists because it could explain some hemorrhagic complications. PMID- 12756502 TI - Early results of a modified technique of cryosurgery. AB - The objective of this study was to present a simple, convenient, and reliable technique for the application of liquid nitrogen and to evaluate the effectiveness of curettage and cryosurgery. Between 1992 and 2002, 24 patients who had benign aggressive and low-grade malignant bone tumors were treated by curettage and cryosurgery. While cryosurgery was applied by the "direct pour" technique in the first seven patients, it was applied by the "pressurized spraying" technique in the others. Functional results were graded according to Enneking. The mean follow-up was 47 (range 9-131) months. There were no local recurrences. Three patients who underwent the direct pour technique developed partial skin necrosis. The functional results were excellent in 14 patients, good in six, and fair in four. When compared with previous reports on cryosurgery and its application techniques, we detected no tumor recurrence or complications, including soft-tissue injury, infection, and late fracture with liquid nitrogen applied by the pressurized spraying technique. PMID- 12756504 TI - Diversification of exogenous genes in vivo in Neurospora. AB - We have adapted the meiotic recombination hotspot cog of Neurospora crassa for shuffling exogenous DNA, providing a means of generating novel genes in situ from sequences introduced into chromosomes. Genes to be diversified are inserted between the his-3 locus and cog. Diversification crosses are heterozygous both for alleles of the exogenous DNA and for auxotrophic alleles of his-3. Progeny selected for ability to grow without histidine supplementation are enriched for exchange events within the exogenous DNA. Exchange events initiated by cog can propagate past DNA sequences mismatched for more than 370 bp and complete exchanges in patches of matched sequence as short as 24 bp, parameters that make the system suited for use in the directed evolution of genes for protein engineering. Here we demonstrate the system by shuffling human immunoglobulin kappa chain genes and also endoglucanase genes derived from different species of fungi. PMID- 12756503 TI - Enhanced washout of 99mTc-tetrofosmin in hypertrophic cardiomyopathy: quantitative comparisons with regional 123I-BMIPP uptake and wall thickness determined by MRI. AB - The diagnostic value of technetium-99m tetrofosmin (TF) washout in hypertrophic cardiomyopathy (HCM) was examined by investigating its relation to the metabolic abnormality depicted by iodine-123 beta-methyl- p-iodophenylpentadecanoic acid (BMIPP) uptake and the left ventricular (LV) myocardial wall thickness as measured by magnetic resonance imaging (MRI). TF washout was evaluated in 31 patients with HCM and 23 normal control subjects using 30-min (early) and 3-h (delayed) TF single-photon emission tomography images. The LV myocardial wall was divided into 19 segments and the percentage TF washout, regional BMIPP uptake and LV wall thickness were measured in each segment. Mean TF washout in the patients with HCM was significantly faster than that in normal control subjects (23.7+/ 5.7 vs 13.4+/-4.1, P<0.0001). In the patients with HCM, TF washout showed an excellent correlation with MRI wall thickness ( r=0.82, P<0.0001) and a good inverse correlation with regional BMIPP uptake ( r=-0.72, P<0.0001). In addition, a good linear correlation was observed between TF uptake and MRI wall thickness in the 19 regional segments. In conclusion, the degree of TF washout corresponds well with the severity of myocardial wall thickness and the degree of metabolic abnormality in patients with HCM. These results suggest that enhanced TF washout might provide additional clinical information regarding metabolic alterations in HCM. PMID- 12756505 TI - Evolution of major histocompatibility complex class I genes in Cetartiodactyls. AB - Previous studies of cattle MHC have suggested the presence of at least four classical class I loci. Analysis of haplotypes showed that any combination of one, two or three genes may be expressed, although no gene is expressed consistently. The aim of this study was to examine the evolutionary relationships among these genes and to study their phylogenetic history in Cetartiodactyl species, including cattle and their close relatives. A secondary aim was to determine whether recombination had occurred between any of the genes. MHC class I data sets were generated from published sequences or by polymerase chain reaction from cDNA. Phylogenetic analysis revealed that MHC class I sequences from Cetartiodactyl species closely related to cattle were distributed among the main cattle gene "groups", while those from more distantly related species were either scattered (sheep, deer) or clustered in a species-specific manner (sitatunga, giraffe). A comparison between gene and species trees showed a poor match, indicating that divergence of the MHC sequences had occurred independently from that of the hosts from which they were obtained. We also found two clear instances of interlocus recombination among the cattle MHC sequences. Finally, positive natural selection was documented at positions throughout the alpha 1 and 2 domains, primarily on those amino acids directly involved in peptide binding, although two positions in the alpha 3 domain, a region generally conserved in other species, were also shown to be undergoing adaptive evolution. PMID- 12756507 TI - Gorlin's syndrome with a thin corpus callosum and a third ventricular cyst. AB - Gorlin's syndrome (naevoid basal cell carcinoma) is an autosomal dominant tumor predisposition syndrome, classically consists of multiple basal cell carcinomas of the skin, odontogenic keratocyst of the jaw, various skeletal abnormalities, and lamellar falx calcifications. Many associated lesions have been reported. We report a case of Gorlin's syndrome in a 22-year-old man in whom CT and MR images showed unusual findings of the thin corpus callosum and third ventricular cyst. We present a case of this syndrome with special emphasis on its unusual neuroradiological findings and radiological management. PMID- 12756506 TI - Mutations in the HLA class II genes leading to loss of expression of HLA-DR and HLA-DQ in diffuse large B-cell lymphoma. AB - Loss of expression of human leukocyte antigen (HLA) class II molecules on tumor cells affects the onset and modulation of the immune response through lack of activation of CD4+ T lymphocytes. Previously, we showed that the frequent loss of expression of HLA class II in diffuse large B-cell lymphoma (DLBCL) of the testis and the central nervous system (CNS) is mainly due to homozygous deletions in the HLA region on chromosome band 6p21.3. A minority of cases showed hemizygous deletions or mitotic recombination, implying that mutation of the remaining copy of the class II genes might be involved. Here, we studied three DLBCLs with loss of HLA-DQ expression for mutations in the DQB1 and DQA1 genes and three tumors with loss of HLA-DR expression for mutations in the DRB1 and DRA genes. In one case, a point mutation in exon 2 of the DQB1 gene, leading to the formation of a stop codon, was detected at position 47. In a second case, a stop codon was found at position 11 due to a deletion of 19 bp in exon 1 of the DRA gene. No mutations were found in the promoter sequences of the DRA, DQA1 and DQB1 genes. We conclude that both homozygous deletions and hemizygous deletions or mitotic recombination with mutations of the remaining allele may lead to loss of expression of the HLA class II genes, which is comparable to the mechanisms affecting HLA class I expression in solid cancers. PMID- 12756508 TI - Isotropic apparent diffusion coefficient mapping of postnatal cerebral development. AB - Diffusion-weighted imaging (DWI) allows us to image the motion of tissue water. This has been used to demonstrate acute ischaemia. Diffusion imaging is also sensitive to water movement along neuronal tracts. Our objective was to map brain maturation in vivo using maps of apparent diffusion coefficient (ADC). We studied 22 children without neurological disease aged between 2 and 720 days. MRI was performed at 1.5 tesla. Multislice single-shot echoplanar DWI was performed at b 0 and 1000 s/mm(2). ADC maps were generated automatically and measurements were performed in the basal ganglia, frontal and temporal white matter and the pons. There was a decrease over time in water diffusion in the areas examined, most marked in the frontal (0.887-1.898 x 10(-3) mm(2)/s) and temporal (1.077-1.748 x 10(-3) mm(2)/s)lobes. There was little change, after an initial decrease, in the basal ganglia (0.690-1.336 x 10(-3) mm(2)/s). There was a difference in water diffusion between the anterior (0.687-1.581 x 10(-3) mm(2)/s) and posterior (0.533-1.393 x 10(-3) mm(2)/s) pons. These changes correlate well with those observed in progressive myelination: the increased water content probably reflects incomplete myelination and the decrease with time in water motion reflects the increase in myelinated brain. PMID- 12756509 TI - Effects of atorvastatin, simvastatin, and fenofibrate therapy on monocyte chemoattractant protein-1 secretion in patients with hyperlipidemia. AB - OBJECTIVE: Monocytes that migrate into the arterial wall participate in the development and, eventually, rupture of the atherosclerotic plaque. The aim of this study was to evaluate the secretion of monocyte chemoattractant protein-1 (MCP-1) by monocytes from hyperlipidemic patients treated with hypolipidemic drugs, namely fenofibrate, simvastatin, or atorvastatin to determine what role is played by these drugs in the development and stabilization of the atherosclerotic plaque. METHODS: Fifty-four hyperlipidemic patients, who did not respond to a low fat diet, were treated with fenofibrate, simvastatin, or atorvastatin (18 patients in each group) for 1 month. The control group included 18 normolipidemic, healthy, age-matched participants. Ten hyperlipidemic patients were effectively treated with hypolipidemic diet alone for 1 month. This group was compared with a control group of ten healthy subjects. To accurately evaluate the adhesion molecule levels, we excluded hyperlipidemic patients and control subjects with any inflammatory disease. Before and after treatment, monocytes were isolated from peripheral blood. After stimulation with lipopolysaccharide (LPS), MCP-1 secretion was measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: MCP-1 levels were significantly higher in hyperlipidemic patients than controls: 15.8+/-0.47, 16.7+/-0.23, and 14.9+/-0.45 compared with 12.36+/-0.42 ng/ml. Fenofibrate, atorvastatin, and simvastatin significantly decreased MCP-1 levels from 15.8+/-0.47 to 8.79+/-0.89, from 16.7+/-0.23 to 7.46+/-0.73, and from 14.9+/-0.45 to 10.3+/-0.8 ng/ml, respectively. In the diet-treated group of hyperlipidemic patients, the level of MCP-1 before therapy was significantly higher than in controls (16.89+/-0.31 vs 12.45+/-0.36 ng/ml). The diet therapy caused a significant decrease in levels of MCP-1 to 15.1+/-0.36 ng/ml. There was a correlation between the decreased levels of lipids and the decreased release of MCP-1 in the patients treated with hypolipemic drugs. CONCLUSION: The drug induced decrease in MCP-1 secretion in hyperlipidemic patients suggests that, apart from acting on lipids, the hypolipidemic drugs studied may directly inhibit the activity of monocytes. PMID- 12756510 TI - Effects on postural oscillation and memory functions of a single dose of zolpidem 5 mg, zopiclone 3.75 mg and lormetazepam 1 mg in elderly healthy subjects. A randomized, cross-over, double-blind study versus placebo. AB - OBJECTIVE: In elderly patients, both falls and impaired memory are considerable medical problems. Hypnotics, which are frequently administered to this patient group for the treatment of insomnia, should ideally not impair equilibrium or memory functions. This double-blind, randomised, four-way, cross-over study investigated the effects of frequently prescribed hypnotics from different classes on postural oscillation and memory under real life conditions. Zolpidem 5 mg, zopiclone 3.75 mg, lormetazepam 1 mg (i.e. usual starting doses in elderly) or placebo were administered at night to 48 healthy elderly volunteers aged 65 years or more. The study included four treatment periods separated by wash-out periods of at least 1 week. METHODS: Psychomotor tests up to 9 h or 10 h after drug intake included, for attention and body sway, clinical stabilometric platform (CSP) tests, simple reaction time (SRT), and the critical tracking test (CTT); for memory, the learning memory tasks (LMT) and the Sternberg memory scanning test (mean reaction time [MRT] and percentage of correct answers) were used. For subjective sleep evaluation the Leeds sleep evaluation questionnaire (LSEQ) and for sedation a visual analogue scale (VAS) were used. For safety evaluations, adverse events (AEs) were recorded. RESULTS: The results demonstrate that compared with placebo, the active drugs increased body sway (area eyes open and closed in the CSP); however, this effect disappeared after 5 h with zolpidem, while it disappeared only after 8 h with lormetazepam and zopiclone. All three drugs did not affect attention assessed by the SRT and CTT. Concerning memory, Sternberg MRT at 9 h was not significantly different up to 5 digits for all groups in comparison with placebo, while for 6 digits it was significantly increased with lormetazepam and zopiclone. In the LMT, an impairment of performance was observed with lormetazepam relative to both zolpidem and placebo. CONCLUSION: The safest compared drug with regard to body sway was zolpidem, because of its short-lasting effect. In addition, zolpidem did not show any significant effect on memory functions, in the present dose comparison. PMID- 12756511 TI - CYP3A5*3 and *6 single nucleotide polymorphisms in three distinct Asian populations. AB - OBJECTIVE: To determine the frequencies of two functional single nucleotide polymorphisms, CYP3A5*3 and CYP3A5*6, in the CYP3A5 gene in three distinct Asian ethnic groups, namely, the Chinese, Malays and Indians. METHODS: Single nucleotide polymorphism analyses of CYP3A5*1, *3 and *6 were performed in 296 healthy subjects (108 Chinese, 98 Malays and 90 Indians) using the polymerase chain reaction-restriction fragment length polymorphism method. RESULTS: The *1 allele frequency was 25% in Chinese compared with 40% in Malays and Indians ( P=0.001). The *3 allele frequency was also higher in the Chinese population, being 76% versus 60% in the Malays and Indians ( P=0.001). The Malays and Indians also had allele frequencies significantly different from Caucasian, Japanese and African-American populations (each P0.05). CONCLUSIONS: Daidzein, a principal isoflavone in soybean, in higher doses may inhibit CYP1A2 activity in vivo, and physicians should be aware of potential drug-food interactions. PMID- 12756513 TI - Central and peripheral haemodynamic effects of L-NAME infusion in healthy volunteers. AB - AIMS: To evaluate the effects of the intravenous administration of the nitric oxide synthesis inhibitor N(g)nitro-L-arginine methyl ester (L-NAME) in healthy volunteers. METHODS: L-NAME (0.25, 0.5 and 0.75 mg/kg over 8 min) was infused in 13 healthy male volunteers. Finally, subjects were infused with either L- or D arginine. RESULTS: L-NAME resulted in dose-dependent falls in heart rate 60 bpm (55-64 bpm) to 49 bpm (46-52 bpm) (P<0.01) and increased mean arterial pressure 77.0 mmHg (73.2-80.8 mmHg) to 90.0 mmHg (87.1-92.8 mmHg) (P<0.01). The cardiac output was significantly reduced after each L-NAME infusion, and systemic vascular resistance increased linearly over the dosage range. Cardiac stroke volume was significantly reduced only following 0.75 mg/kg/min L-NAME: from 100 ml (91.3-108.7 ml) to 83 ml (74.7-91.4 ml); P<0.01. Forearm blood flow was unchanged at any dosage. L-arginine but not D-arginine infusion reversed the haemodynamic effects of L-NAME. CONCLUSIONS: Contrasting with the profound dose dependent effects of L-NAME had significant effects on central haemodynamics but no discernible effects on peripheral blood flow. PMID- 12756514 TI - No major difference in inhibitory susceptibility between CYP2C9.1 and CYP2C9.3. AB - OBJECTIVE: CYP2C9 is a polymorphic enzyme, and CYP2C9*3 is associated with decreased metabolic activity. In addition to the impaired metabolism, we investigated whether the CYP2C9*3 exhibited altered inhibitory susceptibility compared with CYP2C9*1. METHOD: In the present study, CYP2C9.1 and CYP2C9.3 were expressed in yeast. Using typical CYP2C9 substrates (diclofenac, tolbutamide and S-warfarin) and a potent CYP2C9 inhibitor (nicardipine), the Ki values for nicardipine on the three metabolisms in CYP2C9*1 and CYP2C9*3 were determined. RESULT: The ratios of Ki(CYP2C9*3)/Ki(CYP2C9*1) on tolbutamide, diclofenac and S warfarin metabolisms were 1.2, 3.1 and 0.8, respectively. CONCLUSION: In conclusion, there are no significant differences in the inhibitory susceptibility between the two CYP2C9 enzymes. PMID- 12756515 TI - Signal that reboxetine use is linked with symptoms of peripheral ischaemia (Raynaud's syndrome). PMID- 12756516 TI - Modulations of early somatosensory ERP components by transient and sustained spatial attention. AB - To investigate when and how spatial attention affects somatosensory processing, event-related brain potentials (ERPs) were recorded in response to mechanical tactile stimuli delivered to the left and right hand while attention was directed to one of these hands. The attended hand either remained constant throughout an experimental block (sustained attention), or was changed across successive trials (transient attention). Attentional modulations of the N140 component and a sustained 'processing negativity' for attended stimuli were observed in both attention conditions. However, attentional effects on earlier somatosensory components differed systematically. Sustained attention resulted in a contralateral negativity overlapping with the N80 component, while transient attention was reflected by a bilateral positivity overlapping with the P100 component. This dissociation indicates that sustained and transient attention affect different somatosensory areas. It is suggested that sustained attention can modulate tactile processing within primary somatosensory cortex (S1), while effects of transient attention are located beyond S1. Overall, results demonstrate that spatial selectivity in touch is mediated by activity modulations in modality-specific somatosensory cortex. PMID- 12756518 TI - Schizophrenia patients' and psychiatrists' perspectives on ethical aspects of symptom re-emergence during psychopharmacological research participation. AB - RATIONALE: Study designs involving medication-free intervals have become the subject of controversy in the current dialogue on the ethics of serious mental illness research. METHODS: Schizophrenia patients ( n=59; response rate 75%; 48% inpatients) and psychiatrists ( n=70; response rate 83%) responded to ten questions about a hypothetical scenario in which a schizophrenia study participant experienced the re-emergence of serious symptoms during the "wash out" phase of a psychopharmacological trial. Patients provided their personal views, and psychiatrists gave their personal views and made predictions as to how schizophrenia patients in general would respond. RESULTS: Schizophrenia patients and psychiatrists judged the hypothetical protocol as moderately harmful. Both expressed relatively low likelihood of willingness to participate in the study, given this potential outcome. Schizophrenia patients and psychiatrists found the decision fairly easy. Psychiatrists underestimated the level of harm and overestimated the difficulty of the decision as perceived by schizophrenia patients. Schizophrenia patients acknowledged that the offer of money and request by their doctor or family would increase the likelihood of their participation, and psychiatrists accurately predicted these responses. In hypothetical decisions about the symptomatic study participant, 38% of patients and 39% of psychiatrists said they would allow him to leave the hospital. A majority of both groups (63% and 52%, respectively) indicated that medication should be given despite the study participant's objection. Psychiatrists incorrectly predicted this response, expecting instead that most schizophrenia patients would support the discharge request and few would support involuntary administration of medication. Patients and psychiatrists offered similar reasons for participation decisions but differed in their strategies for handling the situation. CONCLUSION: These findings suggest potential strengths of decisionally capable schizophrenia patients in assessing ethically important design elements of psychopharmacological trials. Implications for informed consent for research, expectations of the therapeutic obligations of clinical investigators, and the role of psychiatric advance directives in psychopharmacological research are outlined. PMID- 12756517 TI - High-frequency transcranial magnetic stimulation of the supplementary motor area reduces bimanual coupling during anti-phase but not in-phase movements. AB - Previous electrophysiological and neuroimaging studies have provided evidence that the supplementary motor area (SMA) has an important role in the control of bimanual coordination. The present experiment investigated the effects of high frequency repetitive transcranial magnetic stimulation (rTMS) over the SMA region on kinematic variables during cyclical bimanual coordination, with a particular focus on the quality of coordination. Subjects performed metronome-paced trials of in-phase and anti-phase bimanual index-finger movements at near-maximal cycling frequency. During movement execution, rTMS (20 Hz, 0.5 s, 120% hand motor threshold) was applied over one of three positions in the sagittal midline 2.0, 4.0 and 6.0 cm anterior to the primary motor leg area. Sham rTMS was included as a control condition. After rTMS, the mean relative phase error between hands increased, but only in the anti-phase trials. The maximum increase in phase error occurred immediately after rather than during the rTMS train. The effect was largest after stimulation 4 or 6 cm anterior to the leg area of the primary motor cortex. We did not observe any changes in the variability of relative phase or in cycle duration or movement amplitude. Findings are discussed in light of recent functional models on the role of the SMA in bimanual movement control. PMID- 12756519 TI - 2-Butoxyethanol enhances the adherence of red blood cells. AB - We recently presented a unique, chemically-induced rat model of hemolytic anemia and disseminated thrombosis. In this 2-butoxyethanol (BE)-induced model the organs developing infarction are comparable to those seen in human diseases, characterized by hemolysis and thrombosis (e.g., thalassemia, sickle-cell disease, paroxysmal nocturnal hemoglobinuria, disseminated intravascular coagulation, thrombotic thrombocytopenic purpura, and hemolytic uremic syndrome). Red blood cells (RBCs) have special flow properties, namely, self-aggregability, deformability, and potential adherence to endothelial cells (ECs) of the blood vessel wall, which are essential for adequate blood flow and tissue perfusion; their alteration facilitates circulatory disorders. To examine the possible contribution of alterations in RBC flow properties to the observed thrombosis in the present investigation we determined the BE-induced changes in adherence, aggregability, and deformability of RBCs from male and female Fischer F344 rats exposed to two, three, or four daily doses of BE at 250 mg BE/kg body weight. Control animals were treated with the vehicle alone. Blood was taken on days 2, 3, 4, and 29. The administration of BE did not affect the RBCs aggregability but markedly enhanced their adherence to extracellular matrix; such enhancement was correlated with adherence to cultured ECs. RBC/EC interaction has been shown to be a potent catalyst of vascular occlusion in hemolytic hemoglobinopathies; thus the enhanced RBC adherence to EC is a likely mechanism by which thrombosis and organ infarct are induced in BE-treated rats. PMID- 12756520 TI - The effect of quercetin on the mRNA expression of different antioxidant enzymes in hepatoma cells. AB - The flavonol quercetin shows a wide range of effects in biological systems. We investigated whether quercetin exerts its proposed antioxidant properties via the antioxidant enzyme system. Quercetin in a concentration range from 5 to 100 microM decreased manganese superoxide dismutase, glutathione peroxidase, and copper zinc superoxide dismutase mRNA expression levels each by 30-40% in rat hepatoma H4IIE cells. Catalase mRNA expression levels increased about 30% but only with the cytotoxic concentration of 100 microM. Despite the down-regulation of antioxidant enzyme mRNA expression quercetin treatment of cells induced only a mild oxidative stress. Pretreatment of H4IIE cells with quercetin even protected against an oxidative stress resulting from hydrogen peroxide exposure. In conclusion, the antioxidant capacity of quercetin was shown not to be due to the antioxidant enzyme system. PMID- 12756521 TI - An anatomical study of meniscal allograft sizing. AB - Meniscus-to-femoral condyle congruity is essential for the development of circumferential hoop stresses and thus function of the meniscus. When meniscal allograft transplantation is performed using bony anchorage of the insertional ligaments, accurate graft-to-host size matching is therefore essential. The standard method currently employed for size matching of meniscal allografts is to rely on plain radiographs of the host's knee, from which expected meniscal dimensions are measured. This study aimed to examine the correlation between tibial plateau dimensions and meniscal dimensions. We studied 44 donor tibial plateaus with medial and lateral meniscal allografts attached intact. Meniscal and tibial plateau dimensions were measured. Linear regression analysis was used to calculate expected meniscal dimensions from each specimen's plateau dimensions. Using specific medial and lateral tibial plateau width and length measurements, meniscal dimensions could be predicted with a mean error of only 5.0+/-6.4%. When predicting meniscal dimensions from only total bony plateau width, the mean error observed was 6.2+/-8.0%. The difference between the two methods was not statistically significant. The results suggest that meniscal dimensions can be predicted accurately from tibial plateau measurements, with only small mean errors. However, potential size mismatches should be carefully borne in mind by surgeons using meniscal allografts. PMID- 12756522 TI - A clinical case and anatomical study of the innervation supply of the vastus medialis muscle. AB - The innervation supply to the vastus medialis (VM) muscle, a component of quadriceps femoris (QF), is provided by a branch of the femoral nerve (FN) running along the muscle. The course of the nerve from lumbar roots to the muscle has been described by many researchers. It is known to ride along the femoral vein, artery and saphenous nerve and enter the adductor canal (Hunter's canal), and then to divide into branches that supply vastus medialis and the knee joint. Femoral mononeuropathy is uncommon, and is usually due to compression in the spinal level. Hematoma in the psoas and iliacus muscles, drug abuse, lithotomy position and limb lengthening are the other associated reasons for a mononeuropathy of the femoral nerve. Isolated vastus lateralis (VL) atrophies have been reported by a few authors, suggesting that compression of the nerve and direct violation of the nerve with injections might be the reason for mononeuropathy. Isolated VM atrophy has not been previously reported. The purpose of the study was to identify the anatomical structures around the FN branch which innervates the VM muscle. PMID- 12756523 TI - Cardiac allograft vasculopathy: current concepts and treatment. AB - Cardiac allograft vasculopathy (CAV) remains the leading limiting factor of patient and graft survival after the first post-operative year. The pathogenesis involves both immunological and non-immunological factors. Here, we present recent advances and discuss potential preventative and treatment regimens. A review of the current literature of heart transplantation, detailing molecular mechanisms, pharmacological risk factors and novel immunosuppression regimens was performed. Recent findings demonstrate the pivotal role of the endothelium, resulting in release of pro-fibrotic cytokines, recruitment of circulating leucocytes, proliferation of vascular smooth muscle cells, and deposition of extracellular matrix proteins (ECMs). The role of HMG-CoA reductase inhibitors and anti-hypertensives remains controversial, but there is increasing evidence advocating their prophylactic use. We can conclude that novel immunosuppressive agents such as rapamycin, mycophenolate mofetil and FTY-720 are experimental immunosuppressive agents that are undergoing evaluation in clinical trials. The prophylactic use of statins and anti-hypertensive drugs needs to be defined but needs to suggest potential strategies to prolong cardiac allograft survival. PMID- 12756524 TI - Nutrient conservation increases with latitude of origin in European Pinus sylvestris populations. AB - Nutrient availability varies across climatic gradients, yet intraspecific adaptation across such gradients in plant traits related to internal cycling and nutrient resorption remains poorly understood. We examined nutrient resorption among six Scots pine (Pinus sylvestris L.) populations of wide-ranging origin grown under common-garden conditions in Poland. These results were compared with mass-based needle N and P for 195 Scots pine stands throughout the species' European range. At the common site, green needle N (r(2)=0.81, P=0.01) and P (r(2)=0.58, P=0.08) concentration increased with increasing latitude of population origin. Resorption efficiency (the proportion of the leaf nutrient pool resorbed during senescence) of N and P of Scots pine populations increased with the latitude of seed origin (r(2) > or = 0.67, P < or = 0.05). The greater resorption efficiency of more northerly populations led to lower concentrations of N and P in senescent leaves (higher resorption proficiency) than populations originating from low latitudes. The direction of change in these traits indicates potential adaptation of populations from northern, colder habitats to more efficient internal nutrient cycling. For native Scots pine stands, results showed greater nutrient conservation in situ in cold-adapted northern populations, via extended needle longevity (from 2 to 3 years at 50 degrees N to 7 years at 70 degrees N), and greater resorption efficiency and proficiency, with their greater resorption efficiency and proficiency having genotypic roots demonstrated in the common-garden experiment. However, for native Scots pine stands, green needle N decreased with increasing latitude (r(2)=0.83, P=0.0002), and P was stable other than decreasing above 62 degrees N. Hence, the genotypic tendency towards maintenance of higher nutrient concentrations in green foliage and effective nutrient resorption, demonstrated by northern populations in the common garden, did not entirely compensate for presumed nutrient availability limitations along the in situ latitudinal temperature gradient. PMID- 12756525 TI - Stable isotopes as indicators of altitudinal distributions and movements in an Ecuadorean hummingbird community. AB - Altitudinal migration and dispersal is an important component of the life history of several temperate and tropical birds but remains poorly understood due to the limited success of mark and recapture techniques. Stable isotopes of hydrogen (deltaD) in rainfall, and to a lesser extent, carbon (delta13C) in plants are known to change with altitude and hence may provide the basis of a technique for tracking the altitudinal movements in birds and other wildlife. We investigated the potential for this technique by measuring delta13C, deltaD, and delta15N values in tail feathers of eight species of hummingbirds ( Phaethornis malaris, P. syrmatophorus, P. guy, Adelomyia melanogenys, Coeligena torquata, C. lutetiae, Metallura baroni, M. williami) along an altitudinal gradient (300-3,290 m asl) in the Andes Mountains of Ecuador. Feather delta13C and deltaD values were correlated and each changed significantly with elevation above 400 m. In general, we found good agreement between feather deltaD values and those predicted from a generalized relationship of precipitation and surface water deltaD with altitude. Similarly, feather delta13C values showed an enrichment of approximately 1.5 per thousand per 1,000 m over the linear portion of the elevational response. Stable nitrogen isotope values were variable, and so did not provide useful information on elevation in birds, apart from trophic effects. Overall there appears to be good potential for using the (deltaD, delta13C) stable isotope approach to track altitudinal movements and to elucidate previously unrecognized patterns of life history variation in both temperate and tropical species that migrate across elevational isotopic gradients. PMID- 12756526 TI - Preference and performance of a willow-feeding leaf beetle: soil nutrient and flooding effects on host quality. AB - The distribution and abundance of herbivores on plants growing under different environmental conditions may depend upon preference and/or performance. Soil nutrients and water availability are key determinants of herbivore distribution, as both influence plant growth and tissue quality. However, the effects of water on plant quality may depend upon the availability of nutrients and vice versa. Surprisingly few studies have examined the interactions between the two. We investigated the effects of soil nutrient and water availability on (1) the growth and chemistry of the silky willow (Salix sericea Marshall), and (2) the preference and performance of the imported willow leaf beetle (Plagiodera versicolora Laichartig). We conducted two common garden experiments using a similar 2x2 fully factorial design with two levels of soil nutrients (low, high) and two levels of water availability (field capacity, flooded). In the first experiment (larval performance), larval development time and pupal weight were not influenced by nutrient or water availability to the plant. This occurred despite the fact that plants in the high nutrient treatments had higher protein concentration and lower foliar concentrations of the phenolic glycoside 2' cinnamoylsalicortin. In the second experiment (adult preference), we caged four plants (one from each treatment) and released beetles into cages. We found that plant growth and leaf protein depended upon the interaction between nutrient and water availability. Plant growth was greatest in the high nutrient-field capacity treatment and leaf protein was greatest in the high nutrient-flooded treatment. In contrast, adults settled and oviposited preferentially on the high nutrient treatment under flooded conditions, but we found no evidence of interactions between nutrients and water on preference. Thus, at least under flooded conditions nutrients affect adult preference. We also found that foliar protein was correlated positively with adult oviposition preference and per capita egg production. Our results, then, suggest that soil nutrients can influence adult preference, and that adults choose high-quality hosts (high protein) that promote egg production. PMID- 12756527 TI - Effect of pollinator-inflicted ovule damage on floral abscission in the yucca yucca moth mutualism: the role of mechanical and chemical factors. AB - The long-term persistence of obligate mutualisms (over 40 Mya in both fig/fig wasps and yucca/yucca moths) raises the question of how one species limits exploitation by the other species, even though there is selection pressure on individuals to maximize fitness. In the case of yuccas, moths serve as the plant's only pollinator, but eggs laid by the moths before pollination hatch into larvae that consume seeds. Previous studies have shown that flowers with high egg loads are more likely to abscise. This suggests that yucca flowers can select against moths that lay many eggs per flower through selective abscission of flowers; however, it is not known how yucca moths trigger floral abscission. We tested how the moth Tegeticula yuccasella triggers floral abscission during oviposition in Yucca filamentosa by examining the effects of ovipositor insertion and egg laying on ovule viability and floral abscission. Eggs are not laid at the site of ovipositor insertion: we used this separation to test whether wounded ovules were more closely associated with the ovipositor site or an egg's location. Using a tetrazolium stain to detect injured ovules, we determined whether the number of ovipositions affected the number of wounded ovules in naturally pollinated flowers. Two wounding experiments were used to test the effect of mechanical damage on the probability of floral abscission. The types of wounds in these experiments mimicked two types of oviposition-superficial oviposition in the ovary wall and oviposition into the locular cavity-that have been observed in species of Tegeticula. The effect of moth eggs on ovule viability was experimentally tested by culturing ovules in vitro, placing moth eggs on the ovules, and measuring changes in ovule viability with a tetrazolium stain. We found that ovules were physically wounded during natural oviposition. Ovules showed a visible wounding response in moth-pollinated flowers collected 7 12 h after oviposition. Exact location of wounded ovules relative to eggs and oviposition scars, as well as results from the artificial wounding experiments, showed that the moth ovipositor inflicts mechanical damage on the ovules. Significantly higher abscission rates were observed in artificially wounded flowers in which only 4-8% of the ovules were injured. Eggs did not affect ovule viability as measured by the tetrazolium stain. These results suggest that physical damage to ovules caused by ovipositing is sufficient to explain selective fruit abscission. Whether injury as a mechanism of selective abscission in yuccas is novel or a preadaptation will require further study. PMID- 12756528 TI - Extracellular matrix of different composition supports the various splenic compartments of guinea fowl ( Numida meleagris). AB - The extracellular matrix (ECM) of the spleen of the guinea fowl has been studied by immunohistochemistry. Each splenic compartment contains a different composition of ECM. Reticular-fiber-specific collagen III is expressed by the red pulp and thymus-dependent periarterial lymphatic sheath, but silver impregnation reveals a reticular-fiber-like structure in the periellipsoidal white pulp (PWP) where collagen III is absent. The penicillar capillaries of one central artery are enveloped by a single branching sheath or sleeve: the ellipsoid or Schweigger Seidel sheath. The shape of the sleeve shows more resemblance to a deer antler than to an ellipsoid; it emerges at the beginning of the penicillar capillaries and ends at the edge of the PWP. It is wrapped by a novel discontinuous basement membrane-like structure that expresses tenascin and that is named the capsule of the Schweigger-Seidel sheath (CSS). The cuboidal-shaped inner cell layer of the ellipsoid can be identified by a novel monoclonal antibody: BID3. BID3-positive stellate-shaped cells also occur in the PWP, suggesting that this cell population has migratory capability. Monoclonal antibody KIF8 recognizes an ECM component in the ellipsoid not only of guinea fowl, but also of chicken and quail, and may thus identify an ellipsoid-specific antigen. Collagen I is associated with both the basement membrane of the penicillar capillary and the CSS, whereas collagen III is present only in the CSS. Laminin is expressed in the red pulp, but its staining pattern does not indicate the presence of the "ring fibers", which suggests the absence of sinuses. Fibronectin is the only ECM molecule studied that occurs in every splenic compartment, indicating extensive intrasplenic cell migration. PMID- 12756529 TI - Quantitative cDNA-AFLP analysis for genome-wide expression studies. AB - An improved cDNA-AFLP method for genome-wide expression analysis has been developed. We demonstrate that this method is an efficient tool for quantitative transcript profiling and a valid alternative to microarrays. Unique transcript tags, generated from reverse-transcribed messenger RNA by restriction enzymes, were screened through a series of selective PCR amplifications. Based on in silico analysis, an enzyme combination was chosen that ensures that at least 60% of all the mRNAs were represented by an informative sequence tag. The sensitivity and specificity of the method allows one to detect poorly expressed genes and distinguish between homologous sequences. Accurate gene expression profiles were determined by quantitative analysis of band intensities, and subtle differences in transcriptional activity were revealed. A detailed screen for cell cycle modulated genes in tobacco demonstrates the usefulness of the technology for genome-wide expression analysis. PMID- 12756530 TI - Importance of transmembrane segments in Escherichia coli SecY. AB - To assess the functional importance of the transmembrane regions of SecY, we constructed a series of SecY variants, in which the six central residues of each transmembrane segment were replaced by amino acid residues from either transmembrane segment 3 or 4 of LacY. The SecY function, as assessed by the ability to complement cold-sensitive secYmutants with respect to their growth and translocase defects, was eliminated by the alterations in transmembrane segments 2, 3, 4, 7, 9 and 10. Among them, those in segments 3 and 4 had especially severe effects. In contrast, transmembrane segments 1, 5, 6, and 8 were more tolerant to the sequence alterations. The purified protein with an altered transmembrane segment 6 retained, in large measure, the ability to support SecA-dependent preprotein translocation in vitro. These results will help us to further understand how the SecYEG protein translocation channel functions. PMID- 12756531 TI - Mutant casein kinase I (Hrr25p/Kti14p) abrogates the G1 cell cycle arrest induced by Kluyveromyces lactiszymocin in budding yeast. AB - Zymocin, a toxic protein complex produced by Kluyveromyces lactis, inhibits cell cycle progression in Saccharomyces cerevisiae. In studying its action, a resistant mutant ( kti14-1) was found to express the tot-phenotype typical of totDelta cells, toxin target (TOT) mutants that are impaired in RNA polymerase II Elongator function. Phenotypic analysis of a kti14-1 tot3Delta double mutant revealed a functional link between KTI14 and TOT/Elongator. Unlike totDelta cells, the kti14-1 mutant is sensitive to the drug methylmethane sulfonate (MMS), indicating that, besides being affected in TOT function, kti14-1 cells are also compromised in DNA repair. Single-copy complementation identified HRR25, which codes for casein kinase I (CKI), as KTI14. Kinase-minus hrr25 mutations (K38A and T176I) conferred zymocin resistance, while deletion of the other yeast CKI genes ( YCK1-3) had no effect. A mutation in KTI14 that truncates the P/Q-rich C terminus of Hrr25p also dissociates MMS sensitivity from zymocin resistance; this mutant is resistant to the toxin, but shows normal sensitivity to MMS. Thus, although kinase-minus mutations are sufficient to protect yeast cells from zymocin, toxicity is also dependent on the integrity of the C-terminal region of Hrr25p, which has been implicated in determining the substrate specificity or localization of Hrr25p. PMID- 12756532 TI - A candidate-gene approach to clone the sorghum Brown midrib gene encoding caffeic acid O-methyltransferase. AB - The brown midrib (bmr) mutants of sorghum have brown vascular tissue in the leaves and stem as a result of changes in lignin composition. The bmr mutants were generated via chemical mutagenesis with diethyl sulfate (DES) and resemble the brown midrib (bm) mutants of maize. The maize and sorghum brown midrib mutants are of particular value for the comparison of lignin biosynthesis across different, yet evolutionarily related, species. Although the sorghum brown midrib mutants were first described in 1978, none of the Brown midrib genes have been cloned. We have used a candidate-gene approach to clone the first Brown midrib gene from sorghum. Based on chemical analyses of the allelic mutants bmr12, bmr18 and bmr26, we hypothesized that these mutants had reduced activity of the lignin biosynthetic enzyme caffeic acid O-methyltransferase (COMT). After a northern analysis revealed strongly reduced expression of the COMT gene, the gene was cloned from the mutants and the corresponding wild types using PCR. In all three mutants, point mutations resulting in premature stop codons were identified: bmr12, bmr18 and bmr26 are therefore mutant alleles of the gene encoding COMT. RT PCR indicated that all three mutants express the mutant allele, but at much lower levels relative to the wild-type controls. Molecular markers were developed for each of the three mutant alleles to facilitate the use of these mutant alleles in genetic studies and breeding programs. PMID- 12756533 TI - Construction of a mitotic linkage map of Fusarium oxysporum based on Foxy-AFLPs. AB - Construction of the first mitotic linkage map of the asexual fungus Fusarium oxysporum, based on a population of 32 parasexual fusion products, is reported. Molecular markers were developed using a modified AFLP technique which combines a Foxy-specific primer with standard adapter primers. The retroposon Foxy is abundantly present and highly variable in location in F. oxysporum isolates: 43% of the Foxy-AFLP markers tested appeared to be polymorphic between the strains Fol004 and Fol029. Of the 102 Foxy markers obtained, 83 segregated in a 1:1 ratio. The remaining fragments showed a skewed segregation pattern in which the Fol004 derived Foxy fragments were overrepresented. Foxy markers were observed to be clustered, suggesting that active Foxy elements may not transpose very far from their initial insertion sites, or that hotspots for insertion may exist. Linkage analysis revealed 23 linkage groups. Physical linkage between segregating markers predicted to be 20 cM apart was confirmed, indicating that the mitotic linkage map is reliable. PMID- 12756534 TI - A rapid screen for functional mutants of TraM, an autoregulatory protein required for F conjugation. AB - TraM is an autoregulatory protein required for conjugative transfer of the F plasmid. A rapid screening procedure was developed to select for traM mutants constructed by random PCR mutagenesis. The mutated traM gene was cloned into pT7 5, without the traM promoters (collectively called P( traM)), such that these mutants were expressed from the downstream traJ promoter, resulting in constitutive, low-level, transcription of traM by polymerases that had circumnavigated the plasmid. P( traM) was cloned into pPR9tt as a translational fusion in which a DNA fragment containing P( traM), the ribosome binding site and first 24 codons of traM was fused to the 5' end of lacZ. To downregulate beta galactosidase expression, a -1 frameshift mutation was introduced at the junction between traM and lacZ in the fusion. Selected TraM mutants were further characterized for their intracellular levels, electrophoretic mobility on nondenaturing gels, and activity in F conjugation. Point mutations throughout TraM were found to affect both autoregulation and conjugative function. PMID- 12756536 TI - Mapping of gene-specific markers on the genetic map of chickpea (Cicer arietinum L.). AB - With the exception of the fact that it is made up of eight different chromosomes, the physical organization of the 738-Mb genome of the important legume crop chickpea (Cicer arietinum L.) is unknown. In an attempt to increase our knowledge of the basic structure of this genome, we determined the map positions of a series of genes involved in plant defence responses (DR) by genetic linkage analysis. Exploiting the sequence data available in GenBank, we selected genes known to be induced in chickpea and other plants by pathogen attack. Gene specific primers were designed based on conserved regions, and used to detect the corresponding gene sequences in a segregating population derived from an interspecific cross between Cicer arietinum and C. reticulatum. Forty-seven gene specific markers were integrated into an existing map based on STMS, AFLP, DAF and other anonymous markers. The potential of this approach is discussed. PMID- 12756535 TI - The GATE retrotransposon in Drosophila melanogaster: mobility in heterochromatin and aspects of its expression in germline tissues. AB - A full-length copy of the retrotransposon GATE was identified as an insertion in the tandemly repeated, heterochromatic, Stellate genes, which are expressed in the testis of Drosophila melanogaster. Sequencing of this heterochromatic GATE copy revealed that it is closely related to the BEL retrotransposon, a representative of the recently defined BEL-like group of LTR retrotransposons. This copy contains identical LTRs, indicating that the insertion is a recent event. By contrast, the euchromatic part of the D. melanogaster genome contains only profoundly damaged GATE copies or fragments of the transposon. The preferential localization of GATE sequences in heterochromatin was confirmed for the other species in the melanogaster subgroup. The level of GATE expression is dramatically increased in ovaries, but not in testes, of spn-E(1) homozygous flies. We speculate that spn-E is involved in the silencing of GATE via an RNA interference mechanism. PMID- 12756537 TI - Cloning and expression of a novel lipase gene from Bacillus sphaericus 205y. AB - A Bacillus sphaericus strain (205y) that produces an organic solvent-tolerant lipase was isolated in Port Dickson, Malaysia. The gene for the lipase was recovered from a genomic library and sequenced. Phylogenetic analysis was performed based on an alignment of thirteen microbial lipase sequences obtained from the NCBI database. The analysis suggested that the B. sphaericus lipase gene is a novel gene, as it is distinct from other lipase genes in Families I.4 and I.5 reported so far. Expression in Escherichia coli under the control of the lacZ promoter resulted in an eight-fold increase in enzyme activity after a 3-h induction with 1 mM IPTG. The crude enzyme thus obtained showed a slight (10%) enhancement in activity after a 30-min incubation in 25% (v/v) n-hexane at 37 degrees C, and retained 90% of its activity after a similar period in 25% (v/v) p xylene. PMID- 12756539 TI - Site-directed mutational analysis of the novel catalytic domains of alpha aminoadipate reductase (Lys2p) from Candida albicans. AB - The alpha-aminoadipate reductase, a novel enzyme in the alpha-aminoadipic acid pathway for the biosynthesis of lysine in fungi, catalyzes the conversion of alpha-aminoadipic acid to alpha-aminoadipic-delta-semialdehyde in the presence of ATP, NADPH and MgCl(2). This reaction requires two distinct gene products, Lys2p and Lys5p. In the presence of CoA, Lys5p posttranslationally activates Lys2p for the alpha-aminoadipate reductase activity. Sequence alignments indicate the presence of all functional domains required for the activation, adenylation, dehydrogenation and alpha-aminoadipic acid binding in the Lys2p. In this report we present the results of site-directed mutational analysis of the conserved amino acid residues in the catalytic domains of Lys2p from the pathogenic yeast Candida albicans. Mutants were generated in the LYS2 sequence of pCaLYS2SEI by PCR mutagenesis and expressed in E. coli BL21 cells. Recombinant mutants and the wild-type Lys2p were analyzed for their alpha-aminoadipate reductase activity. Substitution of threonine 416, glycine 418, serine 419, and lysine 424 of the adenylation domain (TXGSXXXXK, residues 416-424) resulted in a significant reduction in alpha-aminoadipate reductase activity compared to the unmutagenized Lys2p control. Similarly replacement of glycine 978, threonine 980, glycine 981, phenylalanine 982, leucine 983 and glycine 984 of the NADPH binding domain (GXTGFLG, residues 978-984) caused a drastic decrease in alpha-aminoadipate reductase activity. Finally, substitution of histidine 460, aspartic acid 461, proline 462, isoleucine 463, glutamine 464, arginine 465, and aspartic acid 466 of the putative alpha-aminoadipic acid binding domain (HDPIQRD, residues 460-466) resulted in a highly reduced alpha-aminoadipate reductase activity. These results confirm the hypothesis that specific amino acid residues in highly conserved catalytic domains of Lys2p are essential for the alpha-aminoadipate reductase activity. PMID- 12756540 TI - Genetic and physical mapping of Pi5(t), a locus associated with broad-spectrum resistance to rice blast. AB - To gain an understanding of the molecular basis for resistance to rice blast (Magnaporthe grisea), we have initiated a project to clone Pi5(t), a locus associated with broad-spectrum resistance to diverse blast isolates. AFLP-derived markers linked to Pi5(t)-mediated resistance were isolated using bulked segregant analysis of F(2) populations generated by crossing three recombinant inbred lines (RILs), RIL125, RIL249, and RIL260 with the susceptible line CO39. The most tightly linked AFLP marker, S04G03, was positioned on chromosome 9 of the fingerprint-based physical map of Nipponbare, a well-characterized rice genotype. Flanking BAC-based Nipponbare markers were generated for saturation mapping using four populations, the three initial RILs and an additional one derived from a cross between M202 and RIL260. A BIBAC (binary BAC) library was constructed from RIL260. Using these resources Pi5(t) was mapped to a 170-kb interval, and a contiguous set of BIBAC clones spanning this region was constructed. It had previously been suggested that Pi3(t) and Pi5(t) might be allelic, due to their identical resistance spectrum and tight linkage. We therefore compared genomic regions for lines containing Pi3(t) using the Pi5(t)-linked markers. DNA gel-blot analyses indicated that the region around Pi3(t) is identical to that of Pi5(t), suggesting that Pi3(t) and Pi5(t) are the same resistance gene. PMID- 12756541 TI - Investigations on the biology, epidemiology, pathology and control of Tunga penetrans in Brazil: I. Natural history of tungiasis in man. AB - Tungiasis is an important health problem in poor communities in Brazil and is associated with severe morbidity, particularly in children. The causative agent, the female flea Tunga penetrans, burrows into the skin of its host, where it develops, produces eggs and eventually dies. From the beginning of the penetration to the elimination of the carcass of the ectoparasite by skin repair mechanisms, the whole process takes 4-6 weeks. The present study is based on specimens from 86 patients, for some of whom the exact time of penetration was known. Lesions were photographed, described in detail and biopsied. Biopsies were examined histologically and by means of scanning electron microscopy (SEM). Based on clinical, SEM and histological findings, the "Fortaleza classification" was elaborated. This allows the natural history of tungiasis to be divided into five stages: (1) the penetration phase, (2) the phase of beginning hypertrophy, (3) the white halo phase, (4) the involution phase and (5) residues in the host's skin. Based on morphological and functional criteria, stages 3 and 4 are divided into further substages. The proposed Fortaleza classification can be used for clinical and epidemiological purposes. It allows a more precise diagnosis, enables the assessment of chemotherapeutic approaches and helps to evaluate control measures at the community level. PMID- 12756538 TI - Genetic evidence for a role of phospholipase C at the budding yeast kinetochore. AB - Chromosome segregation during mitosis requires kinetochores, specialized organelles that mediate chromosome attachment to spindle microtubules. We have shown previously that in budding yeast, Plc1p (phosphoinositide-specific phospholipase C) localizes to centromeric loci, associates with the kinetochore proteins Ndc10p and Cep3p, and affects the function of kinetochores. Deletion of PLC1 results in nocodazole sensitivity, mitotic delay, and a higher frequency of chromosome loss. We report here that despite the nocodazole sensitivity of plc1Delta cells, Plc1p is not required for the spindle checkpoint. However, plc1Delta cells require a functional BUB1/BUB3-dependent spindle checkpoint for viability. PLC1 displays strong genetic interactions with genes encoding components of the inner kinetochore, including NDC10, SKP1, MIF2, CEP1, CEP3, and CTF13. Furthermore, plc1Delta cells display alterations in chromatin structure in the core centromere. Chromatin immunoprecipitation experiments indicate that Plc1p localizes to centromeric loci independently of microtubules, and accumulates at the centromeres during G(2)/M stage of cell cycle. These results are consistent with the view that Plc1p affects kinetochore function, possibly by modulating the structure of centromeric chromatin. PMID- 12756542 TI - Entamoeba histolytica infections in captive primates. AB - A group based survey on the presence of Entamoeba histolytica and Entamoeba dispar using real-time PCR among 20 species of captive non-human primates was performed after diagnosis of E. histolytica dysentery in a spider monkey ( Ateles belzebuth hybridus). E. histolytica DNA was detected in three species of New World primates and in three species of Old World primates. In five of six E. histolytica isolates, it was possible to amplify the SREHP gene. They all revealed the same pattern after AluI digestion, indicating a common source of infection. E. dispar DNA was detected in two species of New World monkeys and three species of Old World monkeys. The results demonstrate that E. histolytica is capable of causing symptomatic and non-symptomatic infections in Old World and New World non-human primates. To our knowledge, this is the first report of E. histolytica sensu stricto in non-human primates after the redescription separating it from E. dispar in 1993. PMID- 12756543 TI - Purification of Sarcocystis neurona sporocysts from opossum (Didelphis virginiana) using potassium bromide discontinuous density gradient centrifugation. AB - This report describes a new, inexpensive procedure for the rapid and efficient purification of Sarcocystis neurona sporocysts from opossum small intestine. S. neurona sporocysts were purified using a discontinuous potassium bromide density gradient. The procedure provides a source of sporocyst wall and sporozoites required for reliable biochemical characterization and for immunological studies directed at characterizing antigens responsible for immunological responses by the host. The examined isolates were identified as S. neurona using random amplified polymorphic DNA primers and restriction endonuclease digestion assays. This method allows the collection of large numbers of highly purified S. neurona sporocysts without loss of sporocyst viability as indicated by propidium iodide permeability and cell culture infectivity assays. In addition, this technique might also be used for sporocyst purification of other Sarcocystis spp. PMID- 12756544 TI - Structural and functional analysis of an amplification containing a PGPA gene in a glucantime-resistant Leishmania (Viannia) guyanensis cell line. AB - Drug resistance is a complex phenomenon in Leishmania and commonly involves gene amplification. Active efflux and metal sequestration through a P-glycoprotein have been pointed to as the major mechanisms used by drug-resistant Leishmania. A gene amplification from a glucantime-resistant Leishmania (Viannia) guyanensis cell line was characterised in an attempt to understand the mechanism of metal resistance in this pathogenic species. We show that the amplification is present as an extrachromosomal amplicon of 30 kb and contains a PGPA gene ( LgPGPA), which is overexpressed in the resistant line as shown by Northern and Western blot analyses. In addition, we gathered evidence from transfection experiments for the role of the LgPGPA gene in oxyanion resistance in L. (V.) guyanensis. Our work indicates that, in this pathogenic New World Leishmania species, amplification of the PGPA gene is the major determinant in oxyanion resistance. PMID- 12756545 TI - Genetic homogeneity of axenic isolates of Giardia intestinalis derived from acute and chronically infected individuals in Mexico. AB - Twenty-six axenic isolates of Giardia intestinalis, established in Mexico City over an 11-year period from symptomatic and asymptomatic individuals with acute or chronic infections, were typed genetically. A segment of the glutamate dehydrogenase gene was amplified by PCR and examined by restriction analysis using BspH1 and ApaI to determine the major genetic assemblages to which the isolates belonged. This was coupled with the amplification and analysis of segments of variant-specific surface protein genes to determine genetic subgroupings. Despite their heterogeneous clinical backgrounds, the isolates were found to be genetically homogeneous-all belonging to genetic group I of assemblage A. The results show that type A-I G. intestinalis is ubiquitous in Mexico City and that host factors play an important, if not dominant, role in determining the clinical outcome of Giardia infections in humans. PMID- 12756546 TI - Zoonotic genotypes of Blastocystis hominis detected in cattle and pigs by PCR with diagnostic primers and restriction fragment length polymorphism analysis of the small subunit ribosomal RNA gene. AB - Twenty two Blastocystis isolates from cattle and pigs were genotyped by PCR using diagnostic primers, and the homology among isolates was then confirmed by restriction fragment length polymorphism (RFLP) analysis of the small subunit ribosomal RNA gene. Two distinct genotypes, subtypes 1 and 3, were detected in one and two of the ten isolates from cattle, and three and one of the 12 isolates from pigs, respectively. The RFLP profiles of all isolates designated as subtype 1 and subtype 3 were identical to those of known ribodemes 1 and 2, respectively. We found that 31.8% (7/22) of isolates examined were zoonotic genotypes of Bastocystis hominis. However, 68.2% (15/22) of isolates were not amplified with any of the diagnostic primers, and the profiles of these isolates were different from any ribodemes of B. hominis reported previously. This study shows that Blastocystis organisms in cattle and pigs are a potential source of human infection. PMID- 12756547 TI - Evidence of phenotypic differences between resistant and susceptible isolates of Pseudosuccinea columella (Gastropoda: Lymnaeidae) to Fasciola hepatica (Trematoda: Digenea) in Cuba. AB - Isolates of Cuban Pseudosuccinea columella susceptible and non-susceptible to Fasciola hepatica miracidial infection were compared for cellular reaction to miracidial development, shell morphometrics, mantle pigmentation pattern, and egg laying behaviour. At 24 h post-exposure, non-susceptible snails showed an early cellular host reaction to encapsulate and phagocytise the miracidium, whereas viable transforming miracidia were observed in exposed susceptible snails, indicating that non-susceptibility resulted from resistance rather than physiological unsuitability. Susceptible isolates (more than 80% susceptible to infection) showed significantly higher values of the morphometric indices shell width/length and aperture width/length than resistant isolates (100% resistant). Mantle pigmentation pattern was analysed using Visilog software; the cumulative distances between mantle spots were plotted and compared between isolates, showing a clear distinction between groups. Egg-laying behaviour also showed significant differences with susceptible snails preferring to lay their eggs on the container's walls whereas resistant snails preferred to lay their eggs on the bottom of the container. PMID- 12756548 TI - A general survey of the helminth parasites of fish from inland waters in the Fayoum Governorate, Egypt. AB - During the present investigation, 450 fishes belonging to seven genera and seven species were examined for helminth parasites. Of these, 216 fishes (48%) were found to harbour Acanthocephala (14%), cestodes (16.22%), Digenea (10.66%), Monogenea (1.77%), or nematodes (6.22%). The species richness and intensity of infections are given. Results summarized from nine studies on the helminth parasites of fish from inland waters are presented. The incidence of infection is reported and compared in host/parasite lists. PMID- 12756549 TI - Environmental dispersal of Cryptosporidium parvum oocysts and cross transmission in cultured bivalve molluscs. AB - Two commercially valuable mollusc species ( Ostrea edulisand Tapes decussatus) were experimentally contaminated with Cryptosporidium parvum oocysts. A direct immunofluorescent antibody technique and inclusion/exclusion of the fluorogenic vital dye propidium iodide were used to test for the presence and viability of the oocysts, showing that transmission of contamination occurred between coexisting species. There was a decrease in the viability of oocysts in the initially uncontaminated molluscs as well as a large decrease in the number of oocysts retained when dead molluscs were used as the source of contamination. The results show the potentially important role that these molluscs play in spreading contamination in depuration plants and areas where aquatic organisms are cultivated. PMID- 12756550 TI - Effects of conspecifics and heterospecifics on individual worm mass in four helminth species parasitic in fish. AB - Intraspecific and interspecific effects on the growth and body size of helminths are rarely studied in natural situations, yet knowing what determines helminth sizes and thus fecundity is crucial to our understanding of helminth ecology and epidemiology. The determinants of average individual worm mass were investigated in four common species of helminths parasitic in trout, Salmo trutta. In the acanthocephalan Echinorhynchus truttae, there was a negative relationship between the intensity of infection by conspecifics and average individual worm size. However, in the acanthocephalans Pomphorhynchus laevis and Acanthocephalus anguillae and in the cestode Cyathocephalus truncatus, the relationship was positive: individual worms were larger on average when co-occurring with many conspecifics than when co-occurring with very few. In addition, the average mass of individual C. truncatus in a host decreased as the total mass of other helminth species in the same host increased. This interspecific effect involves the whole helminth community, as the combined effect of all other helminth species is a better predictor of reduced mass in C. truncatus than the mass of any other species taken on its own. These results illustrate the importance of considering helminth interactions and helminth growth in a natural setting. PMID- 12756551 TI - NO nerves and their targets in a tapeworm: An immunocytochemical study of cGMP in Hymenolepis diminuta. AB - We studied the pattern of cGMP immunostaining (IS) after stimulation with a nitric oxide donor in the presence of an inhibitor of phosphodiesterase in adult Hymenolepis diminuta. cGMP-IS was detected in the peripheral nervous system, especially in nerve fibres close to the body muscle fibres. cGMP-IS also occurred in terminals beneath the basal lamina of the tegument and between the muscle fibres of the suckers. The pattern of cGMP-IS was compared to that of 5-HT-IS and GYIRFamide-IS. TRITC-conjugated phalloidin was used to stain the musculature. PMID- 12756552 TI - Lactic dehydrogenase virus infection reduces the expulsion of the nematode Nippostrongylus brasiliensis. AB - The interleukin (IL)-13-mediated goblet cell response is the major host effector system involved in the expulsion of Nippostrongylus brasiliensis. Lactic dehydrogenase virus (LDV) induced higher levels of N. brasiliensis egg production compared with controls, but the effect of LDV infection on worm expulsion of, and goblet cell and IL-13 responses to, N. brasiliensis have not been studied. In this study, the effects of LDV infection on these host responses against N. brasiliensis were examined. Mice with chronic LDV infection showed significantly lower worm expulsion rates than non-LDV-infected mice after N. brasiliensis infection, and there were no significant differences in the ratio of female versus male adult worms between control and LDV-infected mice. The number of goblet cells in LDV-infected mice was significantly lower than that in controls. In addition, the levels of IL-13 gene expression in lymph nodes were significantly lower in LDV-infected mice compared with controls. These results suggest that LDV infection reduces the protective immune responses against N. brasiliensis infection by the suppression of IL-13 production. PMID- 12756553 TI - CD4+ and CD19+ splenocytes undergo apoptosis during an experimental murine infection with Taenia crassiceps. AB - The Taenia crassiceps cysticercus is a cestode that naturally and experimentally infects rodents in which it reproduces by budding. In the laboratory, a persistent cellular immunosuppression with a concomitant increasing load of parasites has been observed in experimentally infected BALB/cAnN mice. In this study, enhanced apoptosis was found in spleen cells from 30-day infected mice with a typical "ladder-patterned" DNA fragmentation and an increase in phosphatidylserine expression. A characteristic poly-(ADP-ribose) polymerase cleavage indicates that this cell death is caspase-mediated. Apoptosis was detected in the CD4(+) and CD19(+) splenocytes of infected mice after in vitro stimulation with cysticercal antigens. Considering previous results on the crucial role that CD4(+) T cells play in controlling the extent of infection, apoptosis in this T-lymphocyte subpopulation induced by T. crassiceps cysticerci could be responsible for the immunosuppression that underlies parasite success. PMID- 12756554 TI - Effect of nitric oxide releasing drugs on the intensity of infection during experimental trichinellosis in mice. AB - Nitric oxide (NO) has been shown to play a critical role in various physiological and pathological conditions. Apart from its physiological functions, NO indirectly participates in certain aspects of the pathology of infectious diseases. The aim of this work was to examine the influence of NO-releasing drugs on the intensity of infection in mice infected with Trichinella spiralis. The selected substances were nitroglycerin, isosorbide dinitrate, nitrendipine, sildenafil, and pentaerythritol. These were administered over a prolonged period of time: from the 3rd to the 28th day post-infection. Our study showed that NO administered during trichinellosis may enhance the infection in mice as compared to untreated controls. Thus, treatment with pentaerythritol caused a 44% increase in the intensity of infection relative to the untreated controls, sildenafil a 37% increase, and nitrendipine a 30% increase. This effect may be related to the action of NO on the host's defence mechanisms. PMID- 12756555 TI - A Plasmodium homologue of cochaperone p23 and its differential expression during the replicative cycle of the malaria parasite. AB - The complete gene sequence of a major phosphoprotein from the malaria parasite reveals that it is a homologue to cochaperone p23. This p23 homologue is highly conserved between Plasmodium falciparum and other malaria parasites and exhibits 44% sequence identity with the Schizosaccharomyces pombe p23 homologue. The Plasmodium p23 is a relatively abundant cytoplasmic protein with a molecular mass of 34-36 kDa depending on species. Expression of this 34 kDa protein and its mRNA commences in the early ring stage and continues throughout the trophozoite stage. At the beginning of schizogony there is a decrease in the transcription and translation rates and a decline in the amount of the 34 kDa protein. The exact role of the 34 kDa phosphoprotein in parasite replication and differentiation is not known, but the Plasmodium p23 homologue may play a role in parasite proliferation and differentiation through its interactions with protein kinases and other chaperones. PMID- 12756556 TI - Susceptibility of heat shock protein 70.1-deficient C57BL/6 J, wild-type C57BL/6 J and A/J mice to Trypanosoma congolense infection. AB - The heat shock protein (HSP) 70.1 gene lies on mouse chromosome 17 among the candidates for Tir1, the major quantitative trait locus associated with response to Trypanosoma congolense infection. To evaluate whether the HSP70.1 gene is involved in the response, we compared the susceptibility of HSP70.1-deficient C57BL/6 J, resistant wild-type C57BL/6 J and susceptible A/J mice. No differences were observed between HSP70.1-deficient and wild-type C57BL/6 J mice in survival time, levels of parasitemia and anemia, suggesting that there is no involvement of the HSP70.1 gene in control of T. congolense infection. The course of infection was markedly different between A/J and C57BL/6 J mice. A/J mice showed a bi-phasic survival pattern, which seemed to be associated with two waves of high parasitemia, but developed only moderate anemia. C57BL/6 J mice controlled parasitemia well but developed severe anemia in the late stage of infection. PMID- 12756557 TI - Pneumocystis carinii pneumonia as a complication of bendamustine monotherapy in a patient with advanced progressive breast cancer. AB - BACKGROUND: Bendamustine is an alkylator with anticipated antimetabolic activity. It has shown activity in malignant lymphoma, multiple myeloma, and breast cancer. Recognized side-effects are relatively mild with myelosuppression as the dose limiting toxicity. The CD4/CD8 ratio may be reduced. To what extent the alteration of lymphocytes, especially CD4(+) lymphocytes, correlates with an increase in opportunistic infections cannot be definitively answered. CASE REPORT: The patient, female, aged 48 years, was suffering from an advanced progressive breast cancer. After initial treatment with several chemotherapies, a cytotoxic therapy was initiated, with bendamustine (150 mg/m(2)) administered on two consecutive days and repeated every 4 weeks. After five courses, the patient developed Pneumocystis carinii pneumonia (PCP), disclosed in the bronchoalveolar lavage. While receiving bendamustine therapy, the CD4(+) and CD8(+) lymphocyte counts in the peripheral blood were determined by flow cytometry. The next-to normal CD4/CD8 ratio before therapy (0,82) had decreased to 0,05 during the therapy mainly due to a decline of CD4(+) lymphocyte. The patient was seronegative for human immunodeficiency virus. In spite of high-dose intravenous trimethoprim/sulfamethoxazole and methylprednisolone application, the patient died of a respiratory failure 3 days after PCP was diagnosed. CONCLUSION: Bendamustine is capable of inducing a reduction in CD4(+) lymphocyte counts causing a severe T-lymphocyte-mediated immunosuppression. Measuring CD4(+) lymphocyte counts may be helpful in determining the risk of PCP in patients treated with bendamustine. PMID- 12756559 TI - Regional distribution of risk for childhood diabetes in Austria and possible association with body mass index. AB - This study was performed to investigate whether the body mass index (BMI) of diabetic children at manifestation is related to the non-random risk distribution in Austria and whether there is an association with the BMI of the background population. All newly diagnosed cases aged <15 years (n=1644) from 1989 to 2000 were allocated to districts using the postal code. BMI at the time of diagnosis was derived from the Austrian incidence data sheet in 99.3% of the registered cases. Data on BMI of the whole population >14 years of age for the year 1991 and birth weight for the years 1989-1999 were derived from Statistics Austria. Linear regression as well as ecological regression (Poisson) models with correction for non-linearity were used. BMI at diabetes manifestation was positively associated with year of diagnosis (P<0.05) and the age at diagnosis (P<0.01). An increase in BMI when moving from West to East (P<0.01) and a higher BMI at diagnosis in boys than girls (P<0.01) was observed. The ecological regression model showed a positive significant association of BMI in newborns and a positive trend of BMI in the background population with incidence rates. CONCLUSION: our data support the hypothesis that obesity may be a risk factor not only for type 2 but for type 1 in children acting as an accelerator for the clinical manifestation. PMID- 12756560 TI - Inpatient treatment of obese children: a multicomponent programme without stringent calorie restriction. AB - This prospective clinical case-control study describes the effect of an inpatient multicomponent treatment programme for obese children and adolescents on their weight and psychological well being. We studied 38 patients and 38 controls on the waiting list, matched for age and gender, referred because of obesity, with a median age of 13 years (range 10-17 years) and a median adjusted body mass index (BMI) of 173% (range 130%-257%). The treatment consisted of a 10-month inpatient programme focussing on attaining a healthy lifestyle by increasing physical activity and offering a healthy diet within a cognitive-behavioural framework. At base line, at the end of the treatment, 6 months and 14 months after completion of the treatment, the adjusted BMI was calculated and psychological variables were measured with the Dutch Eating Behaviour Questionnaire, the Eating Disorder Inventory and the Self-Perception Profile for Children. All patients lost weight during treatment (median -48% of the adjusted BMI, range -4% to -102%), in contrast to the non-treated control subjects (median +6%, range -29% to +27%). The children treated developed a higher self-esteem and were more capable of coping with external eating stimuli. They did not develop anorexia nor bulimia nervosa. At the 6-months follow-up, a median increase in the adjusted BMI of +6% (range -19% to +37%) was found; with an additional increase of +4% (range -30% to +41%) at 14-months follow-up. CONCLUSION: a multicomponent long-term inpatient treatment programme is a valuable treatment option for obese children, with a lasting effect up to 14 months post-treatment. Nevertheless, more research is needed to characterise those children who regain weight after treatment and how this may be prevented. PMID- 12756558 TI - Congenital disorders of glycosylation: review of their molecular bases, clinical presentations and specific therapies. AB - Congenital disorders of glycosylation (CDG, formerly named carbohydrate-deficient glycoprotein syndromes) are a rapidly growing family of inherited disorders affecting the assembly or processing of glycans on glycoconjugates. The clinical spectrum of the different types of CDG discovered so far is variable, ranging from severe multisystemic disorders to disorders restricted to specific organs. This review deals with clinical, diagnostic, and biochemical aspects of all characterized CDGs, including a disorder affecting the N-glycosylation of erythrocytes, congenital dyserythropoietic anemia type II (CDA II/HEMPAS), and the first disorders affecting O-glycosylation. Since the clinical spectrum of symptoms in CDG is variable and may be unspecific, a generous selective screening for the presence of CDG is recommended. PMID- 12756561 TI - The effect of low-dose simvastatin in children with familial hypercholesterolaemia: a 1-year observation. AB - Familial hypercholesterolaemia (FH) is a severe disorder of lipid metabolism associated with an enhanced risk to develop cardiovascular disease later in life, with atherosclerotic lesions beginning already in childhood. These are facts which make an early diagnosis and therapy necessary to prevent or delay such complications. The aim of this study was to investigate the efficacy and safety of low-dose simvastatin, a potent HMG-CoA reductase inhibitor, in children and adolescents with FH. Therefore, 20 children and adolescents (12 females, 8 males) aged between 10 and 17 years with FH were recruited for this 1-year simvastatin study. According to baseline levels of low density lipoprotein (LDL)-C, girls and boys were divided into two groups, one group (with LDL-C <220 mg/dl) starting with a simvastatin dosage of 5 mg/day, the other (with LDL-C >220 mg/dl) 10 mg/day with the possibility to increase dosages up to a daily maximum of 20 mg, if not reaching LDL-C concentrations of <170 mg/dl within the first period. Every 4-8 weeks, weight, height, lipids, Lp(a) and routine safety parameters of all participants were determined by a paediatrician, documenting exactly all side effects. The percentage decrease was 25% for LDL-C in the 5 mg simvastatin period (19% for total cholesterol (tChol)), 30% for LDL-C in the 10 mg period (26% for tChol) and 36% decrease for LDL-C in the 20 mg period (29% for tChol). The changes for high density lipoprotein (HDL)-C were -5.9% (5 mg), +2.9% (10 mg) and -10.9% (20 mg) the percentage decrease in triglycerides was 12.6% (5 mg), 14.3% (10 mg) and 21% (20 mg). The side-effects of simvastatin were of no clinical relevance and all disappeared after a couple of days. CONCLUSION: our results showed that simvastatin seems to be an effective and safe medical therapy even in children and adolescents with familial hypercholesterolaemia. PMID- 12756562 TI - Acellular pertussis and meningococcal C vaccines: cardio-respiratory events in preterm infants. PMID- 12756563 TI - Autosomal recessive polycystic kidney disease: improvement of renal function. PMID- 12756564 TI - Expression of apoptosis-related proteins in endometriomas and benign and malignant ovarian tumours. AB - Apoptosis is a physiological process by which multicellular organisms eliminate superfluous cells. Alterations in apoptosis play a key role in tumour development. The objective was to evaluate the immunohistochemical expression of p53, p21, bax, bak, fas, bcl-2 and bcl-x proteins in 10 endometriomas, 20 benign ovarian tumours (10 mucinous, 10 serous) and 30 malignant ovarian tumours (9 mucinous, 19 serous; 2 endometrioids). p53 positive cells (mean+/-SD) in endometriomas, and benign and malignant tumours were 1.9+/-3.2, 0 and 16.2+/ 33.0, respectively. The difference was significant between benign tumours and endometriomas (P=0.003) but not between endometriomas and malignant tumours. P21 expression in endometriomas and benign and malignant tumours was 19.5+/-27.8, 1.7+/-6.7 and 4.1+/-8.6, respectively. Increased p21 expression was found in endometriomas compared with benign (P=0.001) and malignant (P=0.01) tumours. Bax expression was higher in endometriomas than in benign tumours (P=0.01), but no difference was found between endometriomas and malignant tumours. No difference in bak, fas, bcl-2 or bcl-x expression was observed among the groups. In endometriomas, a negative correlation was found between p53 and fas expression (P=0.04, r=0.66). Although endometriomas have histological features of benign ovarian tumours, endometriomas share with malignant ovarian tumours certain alterations in apoptosis-related proteins. PMID- 12756566 TI - Complementary expression of AP-2 and AP-2rep in ectodermal derivatives of Xenopus embryos. AB - In an attempt to define the pattern of developmental expression of AP-2rep and AP 2 in Xenopus embryos, we cloned a Xenopus AP-2rep cDNA. The AP-2rep message was localized in the organizer region at the gastrula stage whereas AP-2 was expressed ventro-laterally in the animal hemisphere. Later, AP-2rep was expressed in the entire neural tissue at the neurula stage while AP-2 was predominantly expressed in the cranial neural crest areas. The endogenous expression of AP-2 in the neural crest area was diminished by ectopic injection of AP-2rep RNA, suggesting a role for AP-2rep in the differentiation of neural tissues by restricting the expression of AP-2 in the Xenopus embryo. PMID- 12756565 TI - Pregnancy and breast cancer: a population-based study. AB - The incidence of pregnancy-associated breast cancer, i.e. during pregnancy and lactation, and of pregnancy subsequent to a breast-cancer diagnosis will increase as more women choose childbearing at a later age. Few larger series are published on pregnancy-associated breast cancer. In a population-based study, we evaluated the outcome and prognostic factors in 173 breast-cancer patients. One hundred and twenty-two patients had pregnancy-associated breast cancer (20 coincident with pregnancy and 102 during lactation) and 51 patients had pregnancy subsequent to breast cancer. The median follow-up time was 151 months. Histopathological parameters and immunoreactivity for oestrogen and progesterone receptors c-erbB-2 and c-erbB-4 were studied. All three groups had tumours with high histological grade, low frequency of hormone receptors and high expression of c-erbB-2. The pregnancy and lactation groups were near identical with regard to all histopathological parameters and outcome. In the two pregnancy-associated breast cancer groups, tumours were significantly larger, with more extensive lymph-node involvement. For node-negative tumours the respective 5- and 10-year survival rates were 62% and 50% in the pregnancy group and 60% and 50% in the lactation group. For node-positive tumours, respective 5- and 10-year survival rates were 50% and 34% in the pregnancy group and 50% and 33% in the lactation group. In the subsequent group, overall survival was high in both node-negative and -positive groups, with 5- and 10-year survival rates of 80% and 73% and 86% and 76%, respectively. Tumour size, lymph-node status, histological grade, progesterone receptor, oestrogen receptor and c-erbB-2 were significant prognostic factors in the pregnancy-associated breast-cancer patients. PMID- 12756567 TI - Ca2+ signalling and membrane current activated by cADPr in starfish oocytes. AB - Cyclic ADP-ribose (cADPr) is a second messenger that regulates intracellular free [Ca2+] ([Ca2+](i)) in a variety of cell types, including immature oocytes from the starfish Astropecten auranciacus. In this study, we employed confocal laser scanning microscopy and voltage clamp techniques to investigate the source of the cADPr-elicited Ca2+ wave originating from the cortical Ca2+ patches we have described previously. The Ca2+ swing was accompanied by a membrane current with a reversal potential of approximately +20 mV. Decreasing external Na+ almost abolished the current without affecting the Ca2+ response. Removal of extracellular Ca2+ altered neither the Ca2+ transient nor the ionic current, nor did the holding potential exert any effect on the Ca2+ wave. Both the Ca2+ response and the membrane current were abolished when BAPTA, ruthenium red or 8 NH(2)-cADPr were preinjected into the oocytes, while perfusion with ADPr did not elicit any [Ca2+](i) increase or ionic current. However, elevating [Ca2+](i) by uncaging Ca2+ from nitrophenyl- (NP-EGTA) or by photoliberating inositol 1,4,5 trisphosphate (InsP(3)) induced an ionic current with biophysical properties similar to that elicited by cADPr. These results suggest that cADPr activates a Ca2+ wave by releasing Ca2+ from intracellular ryanodine receptors and that the rise in [Ca2+](i) triggers a non-selective monovalent cation current that does not seem to contribute to the global Ca2+ elevation. PMID- 12756568 TI - Liver transplantation in patients with liver cirrhosis and esophageal bleeding. AB - BACKGROUND: Uncontrolled hemorrhage from esophageal varices is one of the most devastating complications of portal hypertension in patients with advanced cirrhosis. METHODS: Drug therapy, endoscopic therapy, transjugular intrahepatic portosystemic shunt (TIPS), or surgical shunts are used with increasing success in the prevention and treatment of bleeding. However, all these treatment modalities have limitations because they do not treat the liver cirrhosis itself. On the other hand, treatment modalities for variceal bleeding may influence the ease of the feasibility of the transplantation procedure. This is particularly the case for surgical treatments like portosystemic shunts and devascularization operations. For this reason these procedures should be avoided if possible. When positioned correctly, a TIPS provides an elegant way of treating portal hypertension without influencing the course of liver transplantation. Liver transplantation offers a treatment that cures both the portal hypertension and the liver disease. However, the use of this method of treatment is limited by the organ availability and by the organ allocation algorithm, resulting in considerable waiting time. CONCLUSION: In conclusion, esophageal bleeding should be noticed as an early warning factor, leading the hepatologist to consider liver transplantation and early listing of the patient. PMID- 12756569 TI - The effect of prolonged submaximal exercise on gas exchange kinetics and ventilation during heavy exercise in humans. AB - This study compared ventilation, gas exchange (oxygen uptake, VO(2)) and the surface electromyogram (EMG) activity of four major lower limb muscles during heavy exercise before (Pre-Ex) and after (Post-Ex) a sustained 90-min cycling exercise at 60% VO(2peak). The 90-min exercise was incorporated under the hypothesis that sustained exercise would alter substrate availability in the second exercise bout causing differences in fibre recruitment patterns, gas exchange and ventilation. Nine trained male subjects [VO(2peak)=60.2 (1.7) ml.kg( 1).min(-1)] completed two identical 6-min bouts of cycling performed at high intensity [approximately 90% VO(2peak); 307 (6) W, mean (SE)]. Ventilation and gas exchange were measured breath-by-breath and the EMG was recorded during the last 12 s of each minute of the two 6-min bouts. EMG signals were analysed to determine integrated EMG (iEMG) and mean power frequency (MPF). VO(2) at min 3 and min 6 in Post-Ex were significantly higher (i.e., +201 and 141 ml.min(-1), respectively, P<0.05) than in Pre-Ex but there was a approximately 25% decrease of the slow component, taken as the difference between min 6 and min 3 [187 (27) vs 249 (35) ml.min(-1), respectively, P<0.05]. The greater whole-body VO(2) after 3 min of exercise in Post-Ex was not accompanied by clear alterations in the iEMG and MPF of the examined leg muscles. Ventilation and heart rate were elevated (approximately 12-16 l.min(-1) and approximately 10 beats.min(-1), respectively, P<0.05) as were the ratios V(E)/O(2) and V(E)/VCO(2) in the Post-Ex tests. It was concluded that the VO(2) and ventilation responses to high-intensity exercise can be altered following prolonged moderate intensity exercise in terms of increased amplitude without associated major changes in either iEMG or MPF values among conditions. PMID- 12756570 TI - The effect of eccentric training at different velocities on cross-education. AB - The purpose of this study was to determine whether cross-education, defined as the increase in strength of an untrained limb after training of the contralateral homologous limb, is specific to low and high velocity eccentric training. Twenty six subjects were randomized into two groups ( n=13 each) that performed unilateral eccentric training of the elbow flexors on an isokinetic dynamometer at velocities of either 30 degrees s(-1) (0.52 rad s(-1)) or 180 degrees s(-1) (3.14 rad s(-1 )). Subjects trained three times per week for 8 weeks. Ten subjects served as controls and did not train. Subjects were tested before and after training for peak torque of the elbow flexors during eccentric and concentric contractions at 30 degrees s(-1) and 180 degrees s(-1). Eccentric peak torque at the velocity of 180 degrees s(-1) in the untrained arm increased only for the group that trained at that velocity (P<0.05). There were no other changes in untrained arms for any of the groups at velocities of 30 degrees s(-1) or 180 degrees s(-1). For the trained arm, the increase in eccentric torque (pooled over velocities) was greatest for the group training at 180 degrees s(-1), whereas the increase in concentric torque was similar for the groups training at 30 degrees s(-1) and 180 degrees s(-1). For the trained arm, there was no specificity for velocity or contraction type. We conclude that cross-education was specific to contraction type and velocity when fast (but not slow) eccentric contractions were used during training; whereas there was no specificity of training in the trained arm. PMID- 12756571 TI - The effects of eccentric and concentric training at different velocities on muscle hypertrophy. AB - The purpose of this study was to examine the effect of isokinetic eccentric (ECC) and concentric (CON) training at two velocities [fast, 180 degrees s(-1 )(3.14 rad s(-1)) and slow,30 degrees s(-1)(0.52 rad s(-1))] on muscle hypertrophy. Twenty-four untrained volunteers (age 18-36 years) participated in fast- ( n=13) or slow- ( n=11) velocity training, where they trained one arm eccentrically for 8 weeks followed by CON training of the opposite arm for 8 weeks. Ten subjects served as controls (CNT). Subjects were tested before and after training for elbow flexor muscle thickness by sonography and isokinetic strength (Biodex). Overall, ECC training resulted in greater hypertrophy than CON training (P<0.01). No significant strength or hypertrophy changes occurred in the CNT group. ECC (180 degrees s(-1)) training resulted in greater hypertrophy than CON (180 degrees s(-1)) training and CON (30 degrees s(-1)) training (P<0.01). ECC (30 degrees s(-1)) training resulted in greater hypertrophy than CON (180 degrees s( 1)) training (P<0.05), but not CON (30 degrees s(-1)) training. ECC (180 degrees s(-1)) training resulted in the greatest increases in strength (P<0.01). We conclude that ECC fast training is the most effective for muscle hypertrophy and strength gain. PMID- 12756572 TI - Effect of sampling strategy on measures of VO2peak obtained using commercial breath-by-breath systems. AB - The purpose of this study was to determine the effect of sampling strategy (i.e., number of breaths) on measured peak rate of oxygen uptake ( VO(2peak)) elicited by a range of severe intensity exercise bouts. The hypothesis was that a smaller sample (i.e., fewer breaths) would produce a higher measure of VO(2peak) and that this effect would be greater in shorter tests than in longer tests. Thirty-three university students performed constant-power cycle ergometer tests at intensities selected to elicit fatigue in ~3.0 min (short duration), approximately 5.5 min (medium duration), and approximately 8.0 min (long duration). Values for VO(2peak) were the highest rates of oxygen uptake obtained using the following sampling methods: single breath, and 3-, 5-, 15- and 30-breath rolling averages. As hypothesized, measures of VO(2peak) increased systematically with decreasing sample size. Contrary to the hypothesis, the effect of sample size was greater in medium duration and long duration tests than in the short duration tests. The interaction between test duration and sample size on measures of VO(2peak) highlights the importance of standardizing the analysis protocol for exercise in the severe domain. If such standardization is not feasible, it should be recognized that specific analysis protocols may exert a substantial effect upon the reported VO(2peak). PMID- 12756573 TI - Do gender differences exist in the ventilatory response to progressive exercise in males and females of average fitness? AB - Gender differences in lung volumes and flow rates, and in respiratory control have been documented previously. How these gender differences affect exercise responses in normal subjects is less clear, particularly as many studies involved highly fit subjects. This study aimed to investigate potential gender differences occurring during progressive exercise in healthy males and females of average fitness. Fourteen males and ten females of mean (SD) age 23 (0.35) years completed a progressive exercise test to exhaustion on a cycle ergometer, with a ramp increase of 15 W min(-1) (female) or 20 W min(-1) (male). All females were studied during the follicular phase of their menstrual cycle. Cardiorespiratory variables were measured, breath by breath, and values were compared at rest, at 40 W, at physiologically equivalent workloads below, at and above the gas exchange threshold and at peak oxygen uptake (VO(2peak)). Mean VO(2peak) (SEM) was 32.4 (2.01) ml kg(-1) min(-1) for the females and 41.9 (1.80) ml kg(-1) min( 1) for the males. Females had a significantly lower end-tidal partial CO(2) pressure at rest and throughout exercise. Increases in exercise minute ventilation were achieved by a significantly greater tidal volume in males, whereas females adopted a significantly greater breathing frequency. Ratings of respiratory discomfort were significantly greater in the male group at physiologically equivalent workloads compared to the female group. This study shows gender differences exist in the ventilatory and sensory response to progressive exercise in untrained subjects. Further work is required to ascertain if these effects are altered during the luteal phase of the menstrual cycle. PMID- 12756574 TI - Complementary neuronal and glial expression of two high-affinity glutamate transporter GLT1/EAAT2 forms in rat cerebral cortex. AB - The glutamate transporter GLT1 is essential in limiting transmitter signaling and restricting harmful receptor overstimulation. It has been shown recently that GLT1 exists in two forms, the generic GLT1 and a 3'-end-spliced variant of GLT1 (GLT1v), both with similar transport characteristics. To differentiate clearly the cellular distribution of both GLT1 forms in the cortex, specific cRNA probes for non-radioactive in situ hybridization were generated and applied to adult rat brain sections. The results were complemented by western and northern blot analyses and by immunocytochemical investigations using specific peptide antibodies against both GLT1 forms. The study confirmed that generic GLT1 mRNA was expressed predominantly in astrocytes and, to a small extent, in neurons, whereas GLT1 protein was detected only in cell membranes of astrocytes. On the other hand, GLT1v mRNA and protein were demonstrated predominantly in neurons and in non-astrocytic glial cells irrespective of the cortical areas studied. A cytoplasmic granular staining of neurons and astrocytes predominated in the demonstration of GLT1v protein. It is concluded that the cellular expression of the two GLT1 forms is complementary. The cytoplasmic vesicular distribution of GLT1v may represent an endogenous protective mechanism to limit glutamate-induced excitotoxicity. PMID- 12756575 TI - Cyclic summation versus m-sequence technique in the multifocal ERG. AB - BACKGROUND: The m-sequence technique is a typical tool for the multifocal ERG. The use of LEDs instead of a computer monitor enables a new technique that merits closer investigation: The cyclic summation technique. The aim of this study was to compare the two methods. METHODS: Six normal right eyes were examined with the RETIscan system using DTL electrodes. With an LED array (display diameter 52 degrees, 103 segments, 1 foveal + 102 arranged in six concentric rings) we studied: (1). first order kernels (m-sequence); (2). 30-Hz flicker responses (m sequence); (3). 30-Hz flicker responses (cyclic summation). The three methods were tested with a pattern of concentric rings generated by selective deactivation of LEDs (the central LED and rings 2, 4 and 6; rings 1, 3 and 5 remained active). In each case six cumulative measurements (40 s each) were made and stored separately. To determine the signal-to-noise ratio, the average mf ERG response to all active LEDs was divided by the average response to the inactive ones. RESULTS: 1. Using cyclic summation the signal-to-noise ratio exceeds the signal-to-noise ratio of both m-sequence-controlled stimuli about twofold. This implies also better spatial resolution with the cyclic summation technique 2. Since the signal-to-noise ratio increases faster with the cyclic summation technique than with the m-sequence technique, the gain of time in mf ERG can reach 80%. CONCLUSION: As far as the signal-to-noise ratio and measuring time is concerned, the cyclic summation technique outmatches the m-sequence technique in mf ERG. PMID- 12756576 TI - The relation between visual acuity, fixation stability, and the size and location of foveal hard exudates after photocoagulation for diabetic maculopathy: a 1-year follow-up study. AB - PURPOSE: To study the relation between changes in visual acuity, fixation stability and the retinal area covered by hard exudates 3, 6 and 12 months after laser photocoagulation for diabetic maculopathy. METHODS: Twenty-four eyes of 24 patients with diabetes mellitus were examined 3, 6 and 12 months after retinal photocoagulation for clinically significant macular oedema. Each examination included visual acuity testing (ETDRS charts), quantification of fixation stability using a scanning laser ophthalmoscope (Rodenstock 101) and fundus photography. The fundus photographs were digitised and the retinal area covered by hard exudates determined by image processing. RESULTS: The retinal area covered by hard exudates decreased significantly during the first 6 months after treatment ( P=0.05, paired t-test), but increased again between 6 and 12 months after treatment. The visual acuity showed an increase, albeit non-significant, during the first 6 months after treatment followed by a decrease between 6 and 12 months after treatment. Quantification of fixation stability showed that four patients with central exudates fixated at the border of these lesions, and in two of these patients the disappearance of the exudates resulted in increased visual acuity and a change in fixation to the former exudate area. CONCLUSIONS: The retinal area covered by hard exudates decreases during the first 6 months after central photocoagulation, but increases again between 6 and 12 months after the treatment. Hard exudates covering the foveal region contribute to disturbance of central vision. PMID- 12756577 TI - Short-wavelength automated perimetry in patients with diabetes mellitus without macular edema. AB - BACKGROUND: The short-wavelength-sensitive (SWS) cone-mediated sensitivity is a sensitive indicator of functional changes of the macula in diabetic maculopathy. This study was performed to investigate whether functional losses of the macula are detectable in patients without a significant macular edema. METHODS: In 45 patients with diabetes mellitus with clear optical media and no macular edema, conventional white-on-white perimetry (WWP) and short-wavelength automated perimetry (SWAP) were performed in the central 10-deg field. Fifty-eight healthy subjects ranging in age from 16 to 62 years served as controls. The two groups did not differ in age. RESULTS: Variance analysis (ANOVA) revealed significantly lower sensitivity in patients with diabetes than in controls. SWAP thresholds were significantly more greatly reduced by diabetes than those of WWP (ANOVA interaction: P=0.003). Post-hoc testing revealed a sensitivity reduction of 2.8 dB ( P=0.0003) in patients with diabetes for SWAP versus 0.46 for WWP ( P=0.15). Subgroup analysis revealed that mean thresholds of SWAP and WWP predominantly were reduced in patients with advanced disease. In patients with no retinopathy, sensitivity was not affected at all. CONCLUSION: SWS sensitivity may be affected in patients with diabetic retinopathy without clinically significant macular edema. Sensitivity loss was pronounced with increasing severity of retinopathy, reflecting the global status of the eye. PMID- 12756578 TI - Pars plana vitrectomy with or without silicone oil endotamponade in post traumatic endophthalmitis. AB - BACKGROUND: Results of core vitrectomy in post-traumatic endophthalmitis are poor. Our initial results of complete vitrectomy with primary silicone oil endotamponade were promising. A comparative study of this procedure with conventional core vitrectomy was therefore carried out. METHODS: A prospective randomized controlled study of 24 consecutive cases of post-traumatic endophthalmitis was conducted. Patients were randomized into two groups in the absence of clinical improvement after primary tap and treatment with intravitreal vancomycin and amikacin: group 1 consisted of patients who underwent core vitrectomy alone, group 2 of patients who underwent complete vitrectomy with silicone oil endotamponade. All patients included in the study received intravenous antibiotics and underwent lensectomy. Patients were followed up 1, 2, 4 and 12 weeks postoperatively. In all patients of group 2, silicone oil was removed 6 weeks after primary surgery. The mean duration of follow-up was 112+/ 55 days. RESULTS: Vision of 20/400 or better was obtained in 58.33% of cases (14/24). Visual acuity of only one patient in group 1 was >or=20/200, compared with that of 58.3% of patients (7/12) in group 2 ( P=0.02). Intra-operative retinal breaks were found in 50% (6/12) of the patients belonging to group 1, but did not affect the final visual outcome. In group 1, 33.33% (4/12) developed rhegmatogenous retinal detachment in the immediate post-operative period. Only one of these patients had useful final visual outcome after resurgery. CONCLUSION: Complete vitrectomy with primary silicone oil endotamponade is a useful treatment modality which improves the anatomical and functional results in post-traumatic endophthalmitis. PMID- 12756579 TI - Simulation of airbag impact on eyes after photorefractive keratectomy by finite element analysis method. AB - BACKGROUND: A simulation model of the human eye which we have developed was applied to simulated airbag ocular injury, to determine the physical and mechanical conditions of the impacting airbag that would cause globe rupture in a post-photorefractive keratectomy (PRK) eye. METHODS: Simulations were performed with a computer using the finite element analysis program PAM-CRASH()(Nihon ESI, Tokyo, Japan). The airbag was set to impact on the surface of post-PRK eyes-D3, D6, D10, and D15-and an intact eye at various impact velocities. Strain on the cornea and sclera exceeding 18.0% and 6.8%, respectively, was assumed to indicate the possibility of rupture of each tissue. RESULTS: In contrast to the intact eye, in post-PRK eyes, at the lowest velocity of 20 m/s, some of the element reached the strain threshold in D15. At the medium velocity of 30 m/s, limited corneal rupture was observed in all situations. At the high velocity, 40 m/s, scleral laceration was found in eyes with all diopters, and apparent corneal rupture was observed in D10 and D15, indicating that globe rupture was very likely to occur. CONCLUSION: These results suggest that severe ocular trauma can be caused in post-PRK eyes by airbags at high impact velocities. Preoperative discussion with candidates for laser refractive surgery regarding the potential for severe ocular injury if the normal integrity of the eye is compromised by surgery may be appropriate. Research on modification of airbag design and deployment to minimize the risk of ocular injury is important. PMID- 12756580 TI - Intrastromal refractive surgery with ultrashort laser pulses: in vivo study on the rabbit eye. AB - BACKGROUND: Femtosecond (fs) laser pulses may offer new possibilities in the field of refractive surgery, especially when using the laser as a microkeratome. By induction of nonlinear absorption processes the laser can be used to perform intrastromal cuts. The conventional microkeratome, associated with numerous potential side effects, can possibly be replaced. Furthermore, refractive lenticules can be prepared within the stroma and removed in a single-step operation. METHODS: In 10 rabbits, cuts were made to create both a lamellar flap and an intrastromal refractive lenticule. The flap was lifted, the lenticule was extracted and, finally, the flap was repositioned (intrastromal laser keratomileusis, ILK). The corneal samples were collected up to 120 days after treatment and processed for histopathological analysis. RESULTS: All flaps could be opened and prepared lenticules could be extracted in one piece by the surgeon. The treated corneas developed a mild wound healing reaction, comparable to that known from excimer laser in situ keratomileusis (LASIK) studies. The wound healing was restricted to the flap-stroma interface, most pronounced at the periphery of the flaps. CONCLUSIONS: The use of the fs-laser offers new possibilities in preparation of corneal flaps, possibly providing advantages over conventional microkeratomes. Furthermore, the fs-laser has the potential to create intrastromal refractive lenticules for complete refractive procedures (ILK). PMID- 12756581 TI - Positive serology for Helicobacter pylori and vomiting in the pregnancy. AB - INTRODUCTION: Recently, several investigators have suggested that H. pylori may be a contributory factor in hyperemesis gravidarum. The purpose of this study was to evaluate whether seropositivity for IgG antibodies to H. pylori may also be related to nausea, vomiting, heartburn and epigastric pain in pregnancy. MATERIALS AND METHODS: One hundred and eighty-five women, at term pregnancy, were included in the study. All women completed a questionnaire regarding information on the number of pregnancies and deliveries, weight gain, smoking and gastrointestinal complaints before and during pregnancy. The presence of H. pylori infection was determined by serology. RESULTS: The overall prevalence rate of H. pylori seropositivity was 45.9%. Women positive for H. pylori IgG were older (28.7+/-4.5 vs. 27.0+/-4.5, p=0.02), had more prior pregnancies (3.2+/-2.1 vs. 2.6+/-1.6, p=0.02) and deliveries (2.6+/-1.6 vs. 2.0+/-1.1, p=0.006) and reported vomiting in the first trimester more frequently than H. pylori negative patients (81.2% vs. 65%, p=0.004). On the other hand vomiting in the second trimester was reported more frequently among smokers during pregnancy compared to non-smokers. CONCLUSIONS: H. pylori seropositivity is significantly associated with emesis gravidarum but not with gastro-intestinal symptoms later in pregnancy. First trimester vomiting more than doubles the likelihood that the gravida is H. pylori IgG positive. PMID- 12756582 TI - High basal estradiol level and FSH/LH ratio in unexplained recurrent pregnancy loss. AB - BACKGROUND: The potential role of diminished ovarian reserve in unexplained recurrent pregnancy loss (RPL) in a retrospective comparative analysis. METHODS: Eighty women with RPL underwent routine work-up to exclude known associations of RPL. Serum FSH, LH and E(2) levels were assessed on the 3rd day of the menstrual cycle. Following investigation, 58 women failed to reveal an identifiable cause and are therefore classified as unexplained RPL. Control group consisted of women in whom the cause of abortions was known such as uterine septum and parental chromosomal abnormalities. Mean age, gravidity, parity, presence of infertility, previous number of miscarriages, duration of marriage were similar in both groups. Day 3 serum levels of FSH, E(2) and FSH: LH ratios were compared in the two groups. RESULTS: Elevated FSH concentrations were equally distributed in the unexplained RPL and control groups. Both day 3 E(2) and FSH:LH ratio were elevated in the unexplained RPL group compared with the control group ( p=0.0066 and p=0.0187 respectively). The percentage of women with elevated FSH and/or E(2) levels on day 3 were significantly higher in the unexplained RPL group than in controls ( p=0.0045). CONCLUSIONS: Unexplained RPL may be associated with diminished ovarian reserve and should be considered in the workup of RPL. PMID- 12756583 TI - Indication of emergency peripartum hysterectomy: review of 17 cases. AB - OBJECTIVES: The objectives were to determine the incidence, indications, associated risk factors and complications with emergency peripartum hysterectomy at King Abdulaziz University Hospital, Saudi Arabia. METHODS: This is a retrospective analysis of 17 cases of emergency peripartum hysterectomy done from January 1, 1991 to December 31, 2002. RESULTS: Seventeen patients of emergency peripartum hysterectomy were identified among 34,379 deliveries and the incidence rate was 0.5 per 1,000. Uterine atony 11 (64.7%, 9 without previa and 2 with previa) and followed by morbid adherent placenta with previa 6 (35.3%, 1 complete placenta accreta and 5 partial adherent placenta) was the most common indication of hysterectomy. Of the atonic group, 3 were primigravidae, 2 of 3 induced and 1 placenta previa. In morbid adherent placenta group the gravidity, previous abortions and prior cesarean deliveries were higher compared to the atonic group and were statistically significant. Conservative surgery performed in 6 (35.3%) patients before proceeding to hysterectomies, 3 (17.7%) patients had uterine artery ligation and 3 (17.7%) internal iliac ligation. Eight (47.1%) hysterectomies were subtotal. Nine (53%) patients developed disseminated intravascular coagulopathy (DIC) and one case (6%) had bilateral ureteric ligation and bladder injury. No maternal deaths occurred. CONCLUSION: Uterine atony still is the leading cause of primary postpartum hemorrhage and the main indications of emergency peripartum hysterectomy. The combination of high parity, cesarean section, prior cesarean delivery and current placenta previa were identified as risk factors, and should alert the obstetrician that an emergency peripartum hysterectomy may needed. Although no maternal mortality occurred morbidity remained high. PMID- 12756584 TI - Complications of in vitro fertilization with intracytoplasmic sperm injection in human immunodeficiency virus serodiscordant couples. AB - INTRODUCTION: Our experience with complications related to in vitro fertilization (IVF), particularly ovarian hyperstimulation syndrome (OHSS) and higher-order multiple gestations, in human immunodeficiency virus (HIV) serodiscordant couples undergoing IVF with intracytoplasmic sperm injection (ICSI) is presented. METHODS: We retrospectively evaluated 132 consecutive IVF-ICSI cycles and the ensuing 43 pregnancies in 74 HIV-seropositive male discordant couples. The diagnosis of moderate and severe OHSS was based on clinical criteria. Higher order multiple gestations were defined as viable triplets or greater by the end of the first trimester verified by ultrasonography. RESULTS: The incidence of moderate and severe OHSS was 4.5% of initiated IVF cycles. Seven of the 43 (16.3%) pregnancies were higher-order multiple gestations (1 quadruplet and 6 triplet pregnancies), resulting in the delivery of 3 sets of triplets and 3 sets of twins, with an ongoing twin pregnancy. Four patients had undergone multifetal pregnancy reduction to twins. There were no seroconversions in the women and the infants. CONCLUSION: HIV serodiscordant couples undergoing fertility treatment with IVF-ICSI should be made particularly aware of the risks of higher-order multiple gestations and OHSS. PMID- 12756585 TI - Expression of p53, bcl-2 and growth hormone receptor in actinic keratosis, hypertrophic type. AB - According to novel investigations, actinic keratosis (AK) is not a premalignant lesion but is a malignant lesion in the evolution to invasive squamous cell carcinoma (SCC). Thus, we analyzed p53, bcl-2 and growth hormone receptor (GHR) expression in hypertrophic-type AK (HAK) to determine the relative importance of these protooncogenes in the biological behavior of HAK. Expression of p53, bcl-2 and GHR was determined by immunohistochemistry in 33 HAK specimens and surrounding perilesional normal skin (PNS). The relative proportions of immunoreactive cells were determined. Of the 33 HAK specimens, 30 (91%) showed immunopositive staining for p53, 33 (100%) for bcl-2, and 12 (36%) for GHR. Highly positive p53 expression in HAK lesions could indicate that p53 mutation is an early and crucial event in lesion development. The detected pattern of the p53/ bcl-2 ratio in HAK suggests an important role for another gene: the proapoptotic gene bax. Our findings indicate that GHR expression could be a biological marker of progression of HAK to SCC. PMID- 12756586 TI - Premature keratinocyte death and expression of marker proteins of apoptosis in human skin after UVB exposure. AB - Epidermal keratinocytes undergo a process of terminal differentiation or cornification that in many aspects resembles apoptosis. It is characterized by the elimination of cell nuclei within the granular layer, whereas the cytoplasm is transformed into horn cells. Premature death of keratinocytes can be induced by extrinsic factors such as UV irradiation. We investigated the time-dependent expression of apoptotic marker proteins in the skin of one healthy human volunteer after irradiation with a fourfold minimal erythema dose (MED) of UVB. The data were supplemented by including healthy skin areas of biopsies from patients UVB-irradiated for therapeutic reasons. Punch biopsies were analysed by in situ end-labelling (ISEL) for DNA strand breaks and by immunohistochemistry for expression of p53, bcl-2, active caspase-3 and its proform, and deoxyribonuclease I (DNase I). Keratinocytes with pyknotic nuclei were first detected 6 h after UVB exposure, and apoptotic keratinocytes (sunburn cells) 12 h after exposure. These aggregated to sunburn bodies after 24 h. In control skin, nuclei with DNA strand breaks were only occasionally detected in the granular layer but 6 h after UVB irradiation in the spinous layer. After 12 h, many sunburn cells were ISEL-positive and positively stained for active caspase-3, P53, and DNase I. Morphometric evaluation of the immunohistochemical data demonstrated that maximal upregulation of P53, DNase I and activation of caspase 3 occurred 12 h after irradiation and in advance of the peak of apoptotic cell death reached after 24 h as verified by ISEL. In contrast, strong Bcl-2 immunostaining appeared restricted to presumed melanocytes and basal cells but was not increased after UVB irradiation. PMID- 12756587 TI - The garlic-derived organosulfur component ajoene decreases basal cell carcinoma tumor size by inducing apoptosis. AB - Although the therapeutic role of ajoene, an organosulfur compound of garlic, in cardiovascular diseases and mycology has been established, its usefulness in cancer treatment has only recently been suggested. We applied ajoene topically to the tumors of 21 patients with either nodular or superficial basal cell carcinoma (BCC). A reduction in tumor size was seen in 17 patients. Immunohistochemical assays for Bcl-2 expression in a selection of these tumors before and after treatment showed a significant decrease in this apoptosis-suppressing protein. On average, the percentage of tumor cells expressing the proliferation marker Ki-67 was not decreased, which suggests that the action of ajoene is not explained by a cytostatic effect. To obtain further insight into the mode of action of ajoene, the BCC cell line TE354T and a short-term primary culture of BCC were analyzed for apoptosis induction after treatment with the drug. Apoptosis was detected by morphology of the cells and by flow cytometry. Ajoene induced apoptosis in a dose and time-dependent manner in these cultures. Taking together the results of the in vivo and in vitro studies, we conclude that ajoene can reduce BCC tumor size, mainly by inducing the mitochondria-dependent route of apoptosis. PMID- 12756588 TI - Stress fractures of the femoral neck and coxa vara. AB - BACKGROUND: Theoretically, coxa vara substantially modifies the biomechanical conditions of the femoral neck, increasing the effect of direct muscle pull and leading to fatigue of opposing muscle groups; such modifications would appear to favour the appearance of stress fractures. METHODS: We studied 22 stress fractures of the femoral neck, 12 in patients with coxa vara (group A) and 10 in patients with a normal neck-shaft angle (group B), to assess the possible influence of the femoral angle in the production of stress fractures. RESULTS: Intergroup differences were found for age at the appearance of the fracture (younger patients in the coxa vara group) and symptom duration (longer in group A). CONCLUSIONS: It is suggested that coxa vara predisposes to femoral neck stress-fracture. PMID- 12756589 TI - Evaluation of diagnostic NOTCH3 immunostaining in CADASIL. AB - CADASIL is caused by mutations in the NOTCH3 gene. Although increasingly recognized as a disease entity, the diagnostic confirmation can be lengthy or inconclusive. Recently, NOTCH3 immunostaining of skin biopsy specimens has been introduced as a new diagnostic test. The aim of this study was to independently assess the diagnostic value of NOTCH3 immunostaining, and determine whether the degree of immunostaining correlates with other disease parameters. We determined NOTCH3 mutation carrier status in 62 symptomatic and asymptomatic individuals from 15 CADASIL families. Skin biopsy specimens of these individuals, as well as of a disease control group, were immunostained with NOTCH3 antibody and blindly analyzed by two independent observers to determine sensitivity and specificity. A semiquantitative NOTCH3 immunostaining score was correlated with clinical, genetic and MRI parameters. The sensitivity was 90.2% and 85.4%, respectively, for the two observers, the specificity 95.2% and 100%; both lower than previously reported. Certain NOTCH3 mutations may underlie false-negative results. False positive results were found in a non-mutated control, and also in one disease control. There was no difference in immunostaining between symptomatic and asymptomatic NOTCH3 mutated individuals. Furthermore, the NOTCH3 immunostaining score did not correlate with clinical or MRI parameters. NOTCH3 immunostaining is a supportive, but not definitive, CADASIL diagnostic test, and should be interpreted in the context of clinical and radiological data. Confirmation by DNA analysis is requisite for positive results, and when there exists high clinical suspicion, also for negative results. PMID- 12756590 TI - Autologous stem cell transplantation for treatment of rectovaginal fistula in perianal Crohn's disease: a new cell-based therapy. AB - BACKGROUND AND AIMS: Rectovaginal fistulas in patients with Crohn's disease are difficult to resolve, and surgical failure is very frequent. Recent studies have shown that adult stem cells extracted from certain tissues, such as adipose tissue, can develop into different tissues, such as muscle. PATIENT AND METHODS: We report here the case of a young patient with Crohn's disease who had a recurrent rectovaginal fistula that was treated by autologous stem-cell transplantation with a lipoaspirate as the source of stem cells. RESULTS: Although Crohn's disease is the worst condition for a surgical approach in cases of rectovaginal fistula, we observed good closure. Since the surgical procedure 3 month ago the patient has not experienced vaginal flatus or fecal incontinence through her vagina. Thus our treatment seems to be effective. CONCLUSION: Cell transplantation to overcome healing problems is a new surgical tool, and careful evaluation of this new modality may provide an opportunity to define a new era in the treatment of surgical challenges associated with healing disorders. Ethical and safety items do not seem to be critical problems using autologous stem cells. PMID- 12756591 TI - Synchronous postoperative adjuvant chemoradiation therapy for locally advanced carcinoma of the rectum. AB - BACKGROUND AND AIMS: The adjuvant management of locally advanced rectal cancer has been the subject of much debate over the past 10 years. Whilst it is now widely accepted that combined chemoradiation therapy is the treatment of choice for adjuvant therapy following resection of high-risk tumours, there is still no clear answer on the sequencing of the two modalities in the postoperative setting. PATIENTS AND METHODS: Soon after the NCI in the United States issued its recommendations on the management of resected rectal cancer, we decided to commence a phase 2 study to collect data prospectively on the toxicity of postoperative combined chemoradiation therapy. Radiation therapy was given early in combination with bolus chemotherapy using 5-fluorouracil ( n=80). The prescribed radiation dose was 50.4 Gy in 28 fractions, and the chemotherapy was 450 mg/m(2) given with fractions 1 - 3 and 26 - 28. On completion of the radiation therapy the patient was given a further four cycles of bolus 5 fluorouracil at monthly intervals. The patients were then closely monitored for side effects from the therapy and for signs of local and distant relapse. RESULTS: Acute toxicity of the therapy was significant, with 16% of patients experiencing severe bowel morbidity. The other major side effects of the therapy were skin reactions, neutropenia and bladder problems. Late bowel toxicity was also severe. The local in field relapse rate was 10%. The majority of relapses were at distant sites, mostly in the liver and lungs. The actuarial survival at 5 years was 55%. CONCLUSION: We conclude that the combined adjuvant postoperative chemoradiation therapy using this protocol is effective but has significant acute and late morbidity. The optimum regimen for those patients requiring postoperative adjuvant therapy is yet to be determined. PMID- 12756592 TI - Caecal volvulus following laparoscopy-assisted sigmoid colectomy for sigmoid volvulus. AB - CASE PRESENTATION. We report a case of caecal volvulus in a patient who underwent laparoscopy-assisted sigmoid resection for sigmoid volvulus 1 year previously. DISCUSSION. Clinico-radiological features and the management of metachronous sigmoid and caecal volvulus are discussed. PMID- 12756593 TI - Alterations in cholinergic and neuropeptide innervation of urinary bladder following partial bladder outlet obstruction. AB - Posterior urethral valves (PUV) are the most common cause of bladder outlet obstruction (BOO) in infancy. Bladder instability, poor compliance and myogenic failure are responsible for the poor long-term prognosis in these patients. Previous studies have reported abundance of sensory neuropeptides, e.g. substance P (SP), calcitonin gene-related peptide (CGRP), vasoactive intestinal polypeptide (VIP) and acetylcholinesterase (AchE) nerves in the urinary bladder. We hypothesized that the functional changes in the bladder following BOO are due to alteration in cholinergic and sensory neuropeptide innervation. We therefore investigated cholinergic and sensory innervation of urinary bladder following BOO. Fifteen immature male guinea pigs (Hartley strain) 3-4 weeks old and weighing approximately 250 g. underwent placement of a silk ligature around the bladder neck to induce BOO. Controls included 5 sham-operated animals. The animals were killed 1, 2 and 4 weeks following obstruction, respectively. Whole mount preparation and conventional sections of bladder wall were performed. AchE histochemistry, and single-label immunofluorescence histochemistry for SP, CGRP and VIP were utilized. Light microscopy and laser scanning confocal microscopy were used to assess the results. AchE staining showed marked increase in cholinergic innervation density within the suburothelial region following BOO. The staining for SP, CGRP and VIP demonstrated marked reduction in sensory nerve density within the suburothelial region 1 week following BOO and the lack of sensory innervation 4 weeks after BOO. The marked reduction in sensory innervation of the bladder and simultaneous increase in cholinergic innervation following BOO may lead to bladder instability and decrease in bladder compliance. PMID- 12756594 TI - Nephron-sparing procedures in 11 patients with Wilms' tumor. AB - PURPOSE: In unilateral Wilms' tumor (WT), tumor nephrectomy is the standard surgical approach, whereas partial nephrectomy (PN) is controversially discussed. The aim of our retrospective study was to show that in selected cases of unilateral WT kidney-sparing operations could be a reasonable alternative to nephrectomy and to discuss the results of patients with bilateral WT treated by tumor enucleation. MATERIALS AND METHODS: From 1981 to 1998, seven patients with unilateral nephroblastoma (four stage I, one stage III and two stage IV) had tumor resection by PN (five right side, two left side), which was planned when the tumor volume was reduced after 4 to 6 weeks of chemotherapy by at least 50%, when the tumor occupied one pole or was easily resectable, when 50% or more of the kidney tissue remained and when paraaortic lymph nodes were free by intraoperative histological examination. In four patients with bilateral WT (stage V) bilateral tumor enucleation was carried out-except in one patient in whom the contralateral kidney had to been removed because of extension of the tumor via the inferior vena cava to the right atrium. All patients ( n = 11) received pre- and postoperative chemotherapy followed by radiotherapy in four patients. RESULTS: All patients with unilateral WT ( n = 7) are still alive and disease free (follow-up time: mean 6.6 years, range: 28 months to 11 years) with normal renal function, although two patients with secondary nephrectomy revealed creatinine clearance levels at the lower range. In six patients primary PN was performed successfully. In a stage III tumor patient (intraperitoneal metastasis, free lymph nodes), secondary nephrectomy was necessary due to renal arterial thrombosis 2 days after PN. In one stage IV tumor patient (lung metastasis, free lymph nodes), the primary resection was not far enough away from the tumor margin so that an additional slice of tissue with then tumor-free margins had to be resected. This patient evolved a local relapse 19 months after PN and had to be nephrectomised thereafter. In the group of bilateral WT patients ( n = 4), one child died 2 months after surgery during chemotherapy because of central venous line sepsis. One patient who additionally suffered from inferior vena cava tumor thrombosis extending to the right atrium making nephrectomy of the right kidney necessary developed chronic renal failure 4.7 years postoperatively. The other two stage V tumor patients have creatinine clearance levels within the normal range. CONCLUSIONS: Kidney-sparing procedures remain the operative approach of choice in patients with bilateral WT, but bear the risk of chronic renal failure when one kidney has to be removed. PN in children with unilateral WT, carried out by an experienced surgeon, is a reasonable alternative to nephrectomy if strict guidelines such as excellent tumor response to preoperative chemotherapy and easy resectability far away from the tumor margins through healthy kidney tissue are followed. Paraaortic lymph nodes must be free of tumor invasion in order to avoid local radiotherapy. PN prevents the patient from having to have dialysis in cases of contralateral nephrectomy resulting from metachronous WT or subsequent renal trauma. PMID- 12756595 TI - Safety-pin ingestion in children: a cultural fact. AB - Pediatric foreign-body (FB) ingestion is a common problem. Many of these FBs are sharp objects such as needles, toothpicks and safety pins (SP). This report reviews the management of SP ingestion in children. During a 16-year period, we recorded 49 pediatric cases of witnessed SP ingestion. In all children, SPs were used to attach the blue beads to the child's suits with the belief of averting the evil eye. The mean age was 8 months ranging from 4 months to 2 years, and 30 patients were males and 19 were females. SPs were most commonly sited in esophagus (37%) and stomach (37%). In the remainder, the SPs have already reached the duodenum and intestine. In this series, 20 (41%) children passed SPs spontaneously, 14 (28.5%) required endoscopic removal and 15 (30.5%) underwent surgery. The outcome of all patients was uneventful. All of the esophageal SPs require endoscopic intervention, however, after passing into stomach the patients can be observed with keeping the surgical intervention in mind if the SP displays a fixed position for more than three days. PMID- 12756596 TI - Histopathological differences between recto-sigmoid Hirschsprung's disease and total colonic aganglionosis. AB - Total colonic aganglionosis (TCA) is a severe form of ultra long Hirschsprung's disease with an incidence of 2 to 14% among all forms of intestinal aganglionosis. C-kit positive interstitial cells of Cajal (ICCs) are pacemaker cells that play a key role in the motility function of the bowel. The aim of this study was to compare the innervation and ICCs distribution in total colonic and recto-sigmoid HD. Full thickness colonic specimens were obtained from four children with TCA, ten with recto-sigmoid HD and four controls. Single immunohistochemistry using peripherin, neuronal nitric oxide synthase (nNOS) and c-kit antibody was performed and analysed in light microscopy. Additionally, whole-mount preparations were stained using anti c-kit immunohistochemistry and NADPH-diaphorase. In the ganglionic bowel of TCA, recto-sigmoid HD and control patients there was a strong nNOS and peripherin immunoreactivity (IR) in ganglia of myenteric and submucous plexus and in thin nerve fibres in the muscle layers. In the TCA there was weak or lack of nNOS IR in the sparse, short nerve trunks of the myenteric and submucous plexuses and muscle layers, whereas nNOS weakly positive nerve trunks were observed in the recto-sigmoid HD bowel. Peripherin IR was markedly reduced in the TCA specimens compared to recto-sigmoid HD. In the TCA specimens there was a lack of ICCs-MY in the smooth muscle layer in all the specimens, whereas in the recto-sigmoid aganglionic bowel ICCs-MY were markedly reduced. Whole-mount preparations showed lack of ICCs-MY and a markedly reduced number of NADPH-positive nerve trunks in TCA. Our findings demonstrate clear histopathological differences between rectosigmoid Hirschsprung's disease and total colonic aganglionosis. PMID- 12756597 TI - Early full blood count and severity of disease in neonates with necrotizing enterocolitis. AB - Thrombocytopaenia occurs during necrotizing enterocolitis (NEC), and nadir platelet count is associated with extent of disease. In paediatric meningococcal disease, the product of neutrophil and platelet count at admission is prognostically useful. We therefore aimed to determine whether the first full blood count (FBC) after diagnosis of NEC is useful as a score for poor outcome and severity of disease. Between 1987 to 2001, neutrophils (N), platelets (P) and their product (PN) was available in 187 neonates treated for NEC at our institution. Neonates with NEC were grouped according to the extent of disease (no gangrene, focal, multifocal and pan-intestinal). Data were not normally distributed so Mann-Whitney U test or analysis of variance (ANOVA) on logged data were used ( p<0.05 was considered significant). Receiver operating characteristics (ROC) curves were used to examine the relationship between specificity and sensitivity. A perfect test would have an ROC curve area of 1. Initial P count and PN product of non-survivors were both significantly lower than in survivors ( p<0.0001), whereas N was not different ( p<0.08). Low Log(10)PN was significantly associated with greater extent of disease (ANOVA; no gangrene vs multifocal, p<0.01, vs panintestinal, p<0.0005), suggesting that the initial FBC could be prognostically useful. Area under the ROC survival curve for neutrophils was 0.58, for platelets 0.75 and for PN product 0.71; thus, although no test performed extremely well, initial platelet count and NP product could be useful in evaluating disease severity in neonates with NEC and for further monitoring. PMID- 12756599 TI - Progression of terminal syrinx in occult spina bifida. PMID- 12756598 TI - Vascular changes play a role in the pathogenesis of necrotizing enterocolitis in asphyxiated newborn pigs. AB - Necrotizing enterocolitis (NEC) is the most common acquired gastrointestinal emergency in neonates. We have developed an animal model of NEC in asphyxiated newborn pigs and investigated the effects of asphyxia on blood flow in superior mesenteric artery and abdominal aorta, cardiovascular data, arterial acid-base and blood gas parameters, and endothelial cytoskeletal structure in mesenteric microvasculature. Anesthetized, mechanically ventilated newborn pigs were included in two groups: piglets underwent severe asphyxia, and sham-operated control animals. A cardiovascular and metabolic failure developed in asphyxiated piglets approximately 1 h after the induction: severe hypotension and bradyarrhythmia were seen and significant reductions of the blood flow were measured in the superior mesenteric artery and abdominal aorta during the critical phase. Rearrangement of cytoskeletal actin structure corresponding to enhanced vascular permeability was seen with bodipy phallacidin in mesenterial endothelium of asphyxiated piglets after a 24-h recovery period. In conclusion, severe vasomotor changes during asphyxia may result in mesenteric endothelial dysfunction implicated in increased vascular permeability, edema formation, and development of NEC in asphyxiated piglets. PMID- 12756601 TI - Delayed recovery of left ventricular regional work after coronary angioplasty in patients with opposite wall old myocardial infarction. AB - To evaluate the changes in left ventricular (LV) regional function during acute ischemia in patients with opposite wall old myocardial infarction (OMI), we examined LV regional work during percutaneous transluminal coronary angioplasty (PTCA) of the left anterior descending artery (LAD) in patients with a posterior OMI. Twelve patients with normal LV contraction (group A) and six patients with posterior OMI (group B) who were scheduled to undergo PTCA were enrolled in this study. All patients had single-vessel coronary artery disease and no collateral circulation. Sixty-second inflation was performed, and data were collected every 10 s. The regional work was calculated from the relationship between the mean wall stress and area strain. Regional work of the interventricular septum decreased after balloon inflation and was at its minimum at the end of inflation (group A: 0.6 +/- 0.3 mJ/cm(3); group B: 0.8 +/- 0.4 mJ/cm(3)). After balloon deflation, the septal regional work increased in both groups, and recovered to baseline at 40 s in group A and at 60 s in group B. Regional work of the posterior wall increased in group A after balloon inflation, but not in group B. The recovery of LV regional work after PTCA is delayed in patients with opposite wall OMI. PMID- 12756602 TI - High-sensitivity C-reactive protein and left ventricular remodeling in patients with acute myocardial infarction. AB - Inflammatory cytokines are suspected to play an important role in the pathophysiology of left ventricular (LV) remodeling. We investigated whether high sensitivity C-reactive protein (CRP) (hs-CRP) is a predictor for LV remodeling in patients with acute myocardial infarction (AMI) with successful reperfusion, and also whether such a situation can be avoided by the administration of angiotensin converting enzyme inhibitors (ACEI) or angiotensin II receptor blockers (ARB). The subjects were 139 patients with an initial attack of anterior myocardial infarction successfully treated by reperfusion therapy. They were randomly divided into the following two groups: an angiotensin (AG) group (91 patients treated with ACEI/ARB) and a NON-AG group (48 patients not treated with ACEI/ARB). Levels of hs-CRP, creatine kinase, human atrial natriuretic polypeptide, brain natriuretic peptide (BNP), fasting blood glucose, serum lipids, fibrinogen, fibrin degradation product, prothromloin time, and activated partial thromboplastin time were measured immediately after 1, 2, 3, and 7 days, and 1 months after the onset of AMI. ACEI or ARB administration lowered hs-CRP levels and prevented the development of LV remodeling. Peak CRP levels significantly correlated with BNP levels during the acute stage (r = +0.54, P < 0.0001), end-diastolic volume index (r = +0.78, P < 0.0001), end-systolic volume index (r = +0.36, P = 0.0405), ejection fraction (r = -0.45, P = 0.0052), left ventricular end-diastolic diameter (r = +0.61, P < 0.0001), cardiac output (r = 0.52, P = 0.0005), cardiac index (r = -0.41, P = 0.0099), and systolic pulmonary arterial pressure (r = +0.48, P = 0.0017) 1 month after the onset of AMI in the NON-AG group but not in the AG group. Logistic multivariate analysis revealed that peak CRP alone was an independent risk factor for the development of LV remodeling in the NON-AG group (odds ratio = 1.79, P = 0.002). These results suggest that hs-CRP is a useful factor for predicting LV remodeling. Furthermore, ACEI or ARB administration to AMI patients showing increased hs-CRP levels during the early stage of the disease could prevent LV remodeling. PMID- 12756603 TI - Antegradely insertable aortic balloon occlusion catheter for aortic arch repair. AB - We have developed an antegradely insertable aortic balloon occlusion catheter for aortic arch repair, and review our experiences of using it. The purpose of the present study was to examine the usefulness of the balloon for surgical treatment of aortic arch aneurysm. In 30 patients with aortic arch aneurysm, including 22 with a non-ruptured and 8 with a ruptured aneurysm, the catheter was antegradely inserted into the descending thoracic aorta through the aortic arch or the aneurysm without opening the pleural space after establishing antegrade selective cerebral perfusion and obtaining cardiac arrest. During distal anastomosis, the catheter occluded the aorta with continuous perfusion of the lower half of the body through an arterial cannula inserted into the femoral artery. Among the patients with a nonruptured aneurysm, two deaths (9.1%) occurred because of aorto broncho-esophageal fistulae or cardiac arrest due to severe asthma attack within 30 days, and the other three hospital deaths were due to aspiration pneumonia, multiple organ failure with preoperative renal dysfunction, or low cardiac output syndrome due to perioperative myocardial infarction. Among the patients with a ruptured aneurysm, three deaths (37.5%) were due to acute myocardial infarction, respiratory failure, or intractable arrhythmia within 30 days, and another hospital death was caused by mediastinitis. No paraplegia was caused in any patient excluding one of the patients with a ruptured aneurysm who could not be weaned from the extracorporeal circulation due to perioperative myocardial infarction. There was no early postoperative serious visceral organ dysfunction except for two patients with postoperative low cardiac output syndrome or preoperative severe renal dysfunction. This catheter was effective in protecting the visceral organs and the spinal cord in the repair of an aortic arch aneurysm. PMID- 12756604 TI - Anomalous origin of the left pulmonary artery from the aorta. Our experience and literature review. AB - Three patients with anomalous origin of the left pulmonary artery (AOLPA) from the aorta underwent surgical correction. Two patients presented with an isolated malformation and one with an associated ventricular septal defect. One of the patients with an isolated malformation presented with an AOLPA from the descending thoracic aorta. Implantation of the AOLPA to the main PA trunk was performed by direct anastomosis in two patients and by interposition of a synthetic graft in the other patient with AOLPA from the descending aorta. All patients survived the operation. Low cardiac output was identified in one patient. The mean residual gradient through the anastomotic site was 21 mmHg at follow-up. We found almost 72 cases with AOLPA reported in the literature with a high mortality rate in patients not undergoing surgery. The most frequently found associated malformation is tetralogy of Fallot and ventricular septal defect. AOLPA from the aorta is a rare but important entity, necessitating a scrupulous preoperative and intraoperative evaluation. Patients with AOLPA can undergo surgical repair with excellent results. PMID- 12756605 TI - Relationship between natural killer activity and anger expression in patients with coronary heart disease. AB - Recently, the contribution of the immune system in the development and progression of arteriosclerosis has been suggested. Moreover, psychological stress has also been reported to affect the immune response. Thus, psychological factors are considered to influence the immune response, and an excessive immune response promotes the progression of arteriosclerosis, resulting in the development of coronary heart disease (CHD). In this study, the relationship between the immune response and psychological factors was investigated in CHD patients (n = 74) and normal controls (n = 64). Natural killer (NK) activity was significantly higher in the CHD group than the normal control (CHD vs normal: 8151.39 vs 6653.06, P < 0.05), and was significantly elevated by the suppression of anger and negative emotions (P < 0.05). These results indicate that the characteristic process of psychological response in CHD patients is related to an overresponse of NK activity. We speculate that this linkage significantly influences the development of CHD. PMID- 12756606 TI - Intramyocardial sustained delivery of basic fibroblast growth factor improves angiogenesis and ventricular function in a rat infarct model. AB - Recently we have demonstrated that the release of basic fibroblast growth factor (bFGF) from a biodegradable gelatin hydrogel carrier depends on the degradation of hydrogel in vivo. The purpose of our study was to assess whether bFGF incorporating gelatin hydrogels induce myocardial angiogenesis and improve left ventricular function in the infarcted myocardium of rats. Studies were conducted in 22 Lewis rats after a 4-week ligation of the proximal left anterior descending coronary artery. The rats were randomized into the following two groups: the control group (n = 11) had an intramyocardial injection of saline alone, and the FGF group (n = 11) had gelatin hydrogel microspheres containing 100 microg of bFGF injected into the border zone of the infarct area after the repeat left thoracotomy. For visualization of the regional myocardial blood flow in the rat heart, (201)Tl images were taken just before and 4 weeks after the treatment using a 4-head single photon emission computed tomography scanner with pinhole collimators. Left ventricular function was also assessed with echocardiography and a micromanometer-tipped catheter. Finally, the extent of myocardial angiogenesis was evaluated quantitatively in the postmortem analysis. The (201)Tl defect score in the control group remained unchanged before and after the treatment, whereas it decreased significantly in the FGF group. Both regional and global left ventricular function was significantly better in the FGF group compared with the control group. The vascular density in the border zone of the infarct in the FGF group was significantly higher than that in the control group. In conclusion, intramyocardial injection of bFGF-impregnated gelatin hydrogels induces functionally significant angiogenesis and improves left ventricular systolic and diastolic function in the infarcted myocardium of rats. PMID- 12756607 TI - Effect of suplatast tosilate, an antiallergic selective Th2 cytokine inhibitor, on acute eosinophilic myocarditis: a case report. AB - We encountered a 44-year-old woman with acute eosinophilic myocarditis that showed positive immunostaining for interleukin-4 in the area of marked infiltration of eosinophils into the myocardium. When prednisolone alone proved ineffective, supplementary treatment with suplatast tosilate, an antiallergic selective Th2 cytokine inhibitor, improved the patient's inflammation, reduced the level of cardiac enzymes, and allowed for a reduction in corticosteroid dosage without any adverse effects. PMID- 12756608 TI - Mitral valve replacement and tricuspid annuloplasty via right mini-thoracotomy in a patient with tracheostomy. AB - This report describes our experience in performing mitral valve replacement and tricuspid annuloplasty via a right mini-thoracotomy in a patient with tracheostomy. A 24-year-old woman was admitted with shortness of breath and palpitations. She had subglottic tracheal stenosis and tracheostomy due to tracheal intubation of long duration. Echocardiography revealed chronic severe mitral and tricuspid valve regurgitation. We planned to perform at first the cardiac, and then the tracheal operation, because her left ventricular function was worsening. To eliminate the potential complications of sternotomy in patients with tracheostomy, we used right mini-thoracotomy. We performed mechanical mitral valve replacement for the mitral valve and De Vega annuloplasty for the tricuspid valve. The patient was transferred to the tracheal surgery clinic after the 20th day. Tracheal resection and anastomosis were performed in this department. Three months later, the patient was asymptomatic. We believe that the right mini thoracotomy approach is a good technique for mitral valve replacement in patients with tracheostomy. PMID- 12756609 TI - A heteroplasmic mitochondrial complex I gene mutation in adult-onset dystonia. AB - Mitochondrial DNA (mtDNA) mutations can cause rare forms of dystonia, but the role of mtDNA mutations in other types of dystonia is not well understood. We now report identification by sequencing, restriction endonuclease analyses, and clonal analyses of a heteroplasmic missense A to G base pair substitution at nucleotide position 3796 (A3796G) in the gene encoding the ND1 subunit of mitochondrial complex I in a patient with adult-onset dystonia, spasticity, and core-type myopathy. The mutation converts a highly conserved threonine to an alanine. The same mutation subsequently was identified in 2 of 74 additional unrelated adult-onset dystonia patients. A muscle biopsy was obtained from 1 of these 2 subjects and this revealed abnormalities of electron transport chain (ETC) activities. The mutation was absent in 64 subjects with early onset dystonia, 82 normal controls, and 65 subjects with Parkinson's disease or multiple system atrophy. The A3796G mutation previously has been reported in 3 of 226 subjects from mitochondrial haplogroup H. Each of the 3 subjects in our study harboring the A3796G mutation was also from haplogroup H. However, a subgroup analysis of haplogroup H subjects from our study indicates that the A3796G mutation is significantly overrepresented among haplogroup H adult-onset dystonia subjects compared with haplogroup H controls (P<0.01). This difference remains significant even after excluding the index patient (P=0.04). These data suggest that, among haplogroup H subjects, the presence of the A3796G mutation increases the risk of developing adult-onset dystonia. PMID- 12756611 TI - Impaired hydrolysis of cisplatin derivatives to aquated species prevents energy dependent uptake in GLC4 cells resistant to cisplatin. AB - It has been widely stated that cisplatin enters cells by passive diffusion, despite some reports supporting a carrier-mediated mechanism. We have determined the rate of uptake of carboplatin (CBDCA), of cisplatin (CDDP) and of aquated forms, at different values of the extracellular pH, in the small lung-cancer cells GLC4 and GLC4/CDDP, cells resistant to CDDP. The rate of CDDP uptake is about 2-fold lower in resistant cells than in sensitive ones; in ATP-depleted cells this rate is about the same for both cell lines. The rate of CBDCA uptake is about 10-fold lower than that of CDDP and is the same in both cell lines independently of the ATP status of the cells. On the other hand, the rate of uptake of the aquated form of CDDP is approximately 40-fold higher than that of CDDP and is the same in both cell lines, but decreases dramatically in ATP depleted cells. The plot of the initial rate of uptake of the aquated species as a function of its extracellular concentration shows a tendency to be saturable with k(m)=1.9 mM. In conclusion, our data show that, in sensitive GLC4 cells, passive diffusion of CDDP, probably in its neutral dichloro form, and active uptake of the aquated form contribute to the platinum uptake. The active transport of CDDP involves at least two steps: (1). the hydrolysis of the dichloro species in a deficient Cl(-) space at the level of the plasma membrane, which is the limiting step, and (2). the active transport of the aquated species. In resistant cells, step (1). should be deficient whereas step (2). is the same as in sensitive cells. For CBDCA this mechanism holds; however, step (1). is so low that the active transport does not contribute to the uptake of CBDCA by cells. PMID- 12756610 TI - WaterScore: a novel method for distinguishing between bound and displaceable water molecules in the crystal structure of the binding site of protein-ligand complexes. AB - We have performed a multivariate logistic regression analysis to establish a statistical correlation between the structural properties of water molecules in the binding site of a free protein crystal structure, with the probability of observing the water molecules in the same location in the crystal structure of the ligand-complexed form. The temperature B-factor, the solvent-contact surface area, the total hydrogen bond energy and the number of protein-water contacts were found to discriminate between bound and displaceable water molecules in the best regression functions obtained. These functions may be used to identify those bound water molecules that should be included in structure-based drug design and ligand docking algorithms. FIGURE The binding site ( thin sticks) of penicillopepsin (3app) with its crystallographically determined water molecules ( spheres) and superimposed ligand (in thick sticks, from complexed structure 1ppk). Water molecules sterically displaced by the ligand upon complexation are shown in cyan. Bound water molecules are shown in blue. Displaced water molecules are shown in yellow. Water molecules removed from the analysis due to a lack of hydrogen bonds to the protein are shown in white. WaterScore correctly predicted waters in blue as Probability=1 to remain bound and waters in yellow as Probability<1x10(-20) to remain bound. PMID- 12756612 TI - Antigenic relationship between five isolates of murine gammaherpesvirus analysed with monoclonal antibodies. AB - A panel of six monoclonal antibodies (mAbs) specific for murine gammaherpesvirus (MHV) was used for analysis of the antigenic relationship between five MHV isolates (MHV 68, MHV 72, MHV 76, MHV 78, MHV S). Two mAbs raised against MHV 72 and four raised against MHV S were used in the study. Antibody-virus interactions were tested using immunochemical (ELISA, Western blot, immunofluorescence) and biological (virus-neutralization) assays. Immunoanalysis by ELISA showed a close antigenic relationship between the five viruses, nevertheless, some antigenic individuality of the isolate MHV S was observed. This isolate originated from a geographically distinct area in Czechia relative to the other four isolates from Slovakia. In Western blot analysis, antibodies to MHV 72 recognized viral antigens with the relative molecular mass about 116,000. Of four mAbs against MHV S, only two reacted with denatured viral antigen in Western blot and showed specificity for the 50-55,000 protein. These findings suggested that both isolates, besides of minor antigenic variability, could differ also in immunodominant proteins. Mabs to MHV S exhibited much stronger virus-neutralizing potency than mAbs to MHV 72, indicating thus that the 50-55,000 antigen might be more relevant for the infectivity of MHV-virus. Immunofluorescence with mAbs allowed specific localization of antigens in virus-infected VERO cells. PMID- 12756613 TI - The partial sequence of RNA 1 of the ophiovirus Ranunculus white mottle virus indicates its relationship to rhabdoviruses and provides candidate primers for an ophiovirus-specific RT-PCR test. AB - A 4018 nucleotide sequence was obtained for RNA 1 of Ranunculus white mottle virus (RWMV), genus Ophiovirus, representing an incomplete ORF of 1339 aa. Amino acid sequence analysis revealed significant similarities with RNA polymerases of viruses in the family Rhabdoviridae and a conserved domain of 685 aa, corresponding to the RdRp domain of those in the order Mononegavirales. Phylogenetic analysis indicated that the genus Ophiovirus is not related to the genus Tenuivirus or the family Bunyaviridae, with which it has been linked, and probably deserves a special taxonomic position, within a new family. A pair of degenerate primers was designed from a consensus sequence obtained from a relatively conserved region in the RNA 1 of two members of the genus, Citrus psorosis virus (CPsV) and RWMV. The primers, used in RT-PCR experiments, amplified a 136 bp DNA fragment from all the three recognized members of the genus, i.e. CPsV, RWMV and Tulip mild mottle mosaic virus (TMMMV) and from two tentative ophioviruses from lettuce and freesia. The amplified DNAs were sequenced and compared with the corresponding sequences of CPsV and RWMV and phylogenetic relationships were evaluated. Assays using extracts from plants infected by viruses belonging to the genera Tospovirus, Tenuivirus, Rhabdovirus and Varicosavirus indicated that the primers are genus-specific. PMID- 12756614 TI - Analysis of the morphogenetic cluster and genome of the temperate Lactobacillus casei bacteriophage A2. AB - The genes that encode the morphogenetic proteins of bacteriophage A2 are clustered and expressed as a single operon which originates a late transcript of more than 20 kb. This DNA stretch is analyzed in the context of the whole phage genome, which presents the following peculiarities: a) the head presents two major proteins that share their NH(2) termini, i.e.: both are translated from a single gene (orf5), b) these two proteins suffer a proteolytic maturation process before being incorporated into the capsid, rendering a 123 NH(2) terminal putative polypeptide that is postulated to be the scaffolding protein of the phage, c) similar maturation processes occur at the portal and tail length determination proteins, having all in common a Pho-Pho-Arg downward arrow sequence (where Pho stands for any hydrophobic amino acid) at the processing point, d) the genes encoding the subunits of the terminase (orf61 and orf2) are separated by the cohesive ends, e) two genes that might mediate lysogenic conversion ( orf19 and orf22) have been identified and f) the genome presents a dispensable region (which covers at least 10 orfs, as judged from analysis of deletion mutants) that might be involved in maintaining its size between the packaging limits of the capsid. PMID- 12756615 TI - Effect of neomycin B on rotavirus plus- and minus-strand RNA synthesis. AB - Rotavirus multiplication was inhibited by neomycin B by affecting the synthesis of the viral genome. Genome replication involves a two-step mechanism, one is plus-strand synthesis or transcription, and the other is minus-strand synthesis or replication. The results indicate that both activities are inhibited by aminoglycoside. The effect on the minus-strand was determined by an in vitro assay using a template-dependant open core preparation. The effect on transcription was explored using transcriptionally active virus particles. In the case of transcription, the inhibitory effect of neomycin B was studied in both initiation and the elongation of the mRNA. Initiation was defined by the synthesis of short transcripts of less than 25 nucleotides in length and elongation as an extension of those molecules into full-length transcripts. The inhibitory effect of neomycin was also mimicked by other aminoglycosides such as lividomycin, paromomycin, and tobramycin. The results may be explained based on the ability of the drug to interact with the stem and loop regions, which, in the case of rotavirus, have been identified at the end of the templates required for both transcription and minus-strand synthesis. PMID- 12756616 TI - Isolation of a new flavivirus related to cell fusing agent virus (CFAV) from field-collected flood-water Aedes mosquitoes sampled from a dambo in central Kenya. AB - Cell fusing agent virus (CFAV) is an RNA insect virus that was isolated from a line of Aedes aegypti mosquito cells and has been assigned to the family Flaviviridae, genus Flavivirus. We report here the first isolation of a CFA-like virus from field-collected mosquitoes. Mosquito larvae and pupae were sampled from flooded dambos in Central Province, Kenya during the short rain season of 1999. Specimens were reared to adults, identified and pooled by species and were tested for the presence of virus. Two virus isolates were obtained from two pools of Aedes macintoshi mosquitoes. The virus isolates replicated only in invertebrate cells in culture and not in vertebrate cells or in mice. The virus isolates did not antigenically cross-react with known arboviruses but were identified to family by reverse-transcriptase polymerase chain reaction (RT-PCR) performed using primers specific to alphaviruses, bunyaviruses and flaviviruses; only the flavivirus-specific primers produced a DNA fragment of the expected size. Nucleic acid sequencing of this fragment showed the two isolates to be nearly identical. Comparison of sequences to the GenBank database using BLAST identified the virus as most closely related to CFAV. Results from cross neutralization tests suggested that, although the BLAST search indicated homology to CFAV, the virus isolated represented a new insect flavivirus. Detailed characterization of this new virus, described in Crabtree et al. [7], further supports this finding. We propose this new flavivirus be designated Kamiti River virus (KRV). This is the first isolation of a CFA-like virus from field-collected mosquitoes and indicates the presence of this group of viruses in nature. PMID- 12756617 TI - Genetic and phenotypic characterization of the newly described insect flavivirus, Kamiti River virus. AB - We have described in the accompanying paper by Sang, et al., ([57], Arch Virol 2003, in press) the isolation and identification of a new flavivirus, Kamiti River virus (KRV), from Ae. macintoshi mosquitoes that were collected as larvae and pupae from flooded dambos in Central Province, Kenya. Among known flaviviruses, KRV was shown to be most similar to, but genetically and phenotypically distinct from, Cell fusing agent virus (CFAV). KRV was provisionally identified as an insect-only flavivirus that fails to replicate in vertebrate cells or in mice. We report here the further characterization of KRV. Growth in cell culture was compared to that of CFAV; although growth kinetics were similar, KRV did not cause the cell fusion that is characteristic of CFAV infection. The KRV genome was found to be 11,375 nucleotides in length, containing a single open reading frame encoding 10 viral proteins. Likely polyprotein cleavage sites were identified, which were most similar to those of CFAV and were comparable to those of other flaviviruses. Sequence identity with other flaviviruses was low; maximum identity was with CFAV. Possible terminal secondary structures for the 5' and 3' non-coding regions (NCR) were similar to those predicted for other flaviviruses. Whereas CFAV was isolated from insect cells in the laboratory, the isolation of KRV demonstrates the presence of an insect-only flavivirus in nature and raises questions regarding potential interactions between this virus and other mosquito-borne viruses in competent vector populations. Additionally, this virus will be an important tool in future studies to determine markers associated with flavivirus host specificity. PMID- 12756618 TI - Cytological modifications in zucchini yellow mosaic virus (ZYMV)-infected Styrian pumpkin plants. AB - The present research demonstrates severe ultrastructural changes induced by zucchini yellow mosaic virus (ZYMV) within the cells of older and younger leaves of Styrian pumpkin plants (Cucurbita pepo L. subsp. pepo var. styriaca GREB.). Cylindrical inclusions (pinwheels), proliferated endoplasmatic reticulum and filamentous viral particles were found throughout the cytoplasm of ZYMV-infected cells and within sieve elements. ZYMV-infection also induced severe modifications in the number and ultrastructure of chloroplasts, whereas mitochondria, nuclei and peroxisomes remained unaffected. A significantly lower number of chloroplasts was observed in all tissues of both ZYMV-infected leaf types when compared to control plants. Statistical quantification revealed that in chloroplasts of ZYMV infected older and younger leaves the amount of plastoglobuli and starch increased significantly, whereas the amount of thylakoids significantly decreased. The present research gives a more precise insight in ZYMV-induced modifications within single cells and organelles, and provides statistical data of the most affected chloroplasts. PMID- 12756619 TI - The morphogenesis of Theiler's murine encephalomyelitis virus is strain specific. AB - During a single cycle infection with the neurovirulent GDVII- and demyelinating DA-strain of Theiler's murine encephalomyelitis virus (TMEV) in L-929 cells, different subviral particles were found for both strains. Early in the assembly process, the DA-strain generated 14 S pentamers composed of the viral proteins VP0, VP1 and VP3, while in GDVII-infected cells, particles with the same protein composition but with a sedimentation coefficient of 20 S were found. These newly discovered 20 S particles are probably virion assembly precursors considering their capsid protein composition and their early time of appearance in infected cells. Near the end of the assembly process, VP0, VP1 and VP3 containing 80 S empty capsids became apparent in GDVII-infected cells, while these particles could not be found in DA-infected cells. The significance of these empty capsids will be discussed. After virion assembly, 14 S particles were observed for both strains. These 14 S particles resulted from the degradation of the 160 S virions as indicated by their protein composition (VP1, VP2, VP3) and time of appearance. Our results demonstrate that the assembly of the GDVII-strain differs from that of the DA-strain. In addition, the strain-specific assembly of TMEV implies that not all picornaviruses assemble as proposed by the poliovirus morphogenesis model and thus rendering its general validity questionable. PMID- 12756620 TI - RT-PCR detection of the expression of the polymerase gene of a novel reptilian herpesvirus in tumor tissues of green turtles with fibropapilloma. AB - An alpha-herpesvirus has recently been associated with green turtle fibropapilloma (FP). To further understand the etiological role of this newfound green turtle herpesvirus (GTHV) in the pathogenesis of FP, expression of GTHV polymerase ( pol) gene was determined in tumors and normal-appearing nontumor tissues and organs from five green turtles suffering multiple fibropapillomas, using reverse transcription-polymerase chain reaction (RT-PCR). Amplification of RNA prepared from tumor tissues evidenced the substantial expression of GTHV DNA pol gene in all specimens tested (15/15). However, GTHV pol gene expression in normal-appearing tissues and organs of affected animals was limited (4/45), and GTHV mRNA was detected only in periorbital tissue (1/2), gall bladder (2/5) and lung (1/5) by nested RT-PCR. By contrast, RT-PCR evaluation of RNA isolated from non-tumored turtles revealed undetectable expression of this herpesvirus gene. cDNA sequence analysis revealed that GTHV gene sequences were identical in different tumors. Our data represent the first evidence of the replication of this putative turtle herpesvirus in affected green turtles and fibropapilloma tissues are always active sites of GTHV mRNA synthesis. These findings extend and substantiate the pathogenic association of GTHV with FP. PMID- 12756621 TI - Sequence analysis demonstrates that Onion yellow dwarf virus isolates from China contain a P3 region much larger than other potyviruses. AB - The complete sequence of an isolate of Onion yellow dwarf virus (OYDV) from Yuhang, Zhejiang province, China, was determined. It was 10538 nts in length and was predicted to encode a polyprotein 3403 amino acids (aa) long with a calculated M(r) of 385.1 kDa. The predicted P3 protein (530 aa) was larger than that of any of the potyviruses sequenced to date (344-378 aa). The additional sequence occurs at the N-terminus of the protein, does not represent a duplication from elsewhere in the OYDV genome and could not be matched to any other sequences in the databases. Similar sequences were found in 4 other Chinese OYDV isolates. Phylogenetic analysis of the amino acid sequences of the polyprotein showed that OYDV is distantly related to Pea seed-borne mosaic virus and the potyviruses of grasses and cereals. PMID- 12756622 TI - In vivo distribution of receptor for ecotropic murine leukemia virus and binding of envelope protein of Friend Murine leukemia virus. AB - Ecotropic infection by Murine leukemia virus (MuLV) infection is initiated by the interaction between the receptor-binding domain of the viral surface glycoprotein (SU) and the cell-surface receptor, mCAT-1. To study the in vivo localization of viral binding site in mice, green fluorescence protein (GFP)-tagged Friend SU (F SU/GFP) was incubated with tissue sections. Lymphohematopoietic organs and a part of the glandular tissues of C3H as well as C57BL/6 mice revealed positive signals for F-SU/GFP binding on the cell surface. In contrast, C4W mice, which is a partial congenic mouse strain carrying the Fv-4 (r) gene on a BALB/c genetic background, exhibited negative signals in most of the organs except for a very weak binding in the pancreas. The expression of mCAT-1 mRNA determined by reverse transcriptase (RT)-polymerase chain reaction (PCR) revealed a similar distribution in C3H, C57BL/6 and C4W mice. Most of the organs including lymphohematopoietic organs and glandular organs revealed significant expression of mRNA for mCAT-1 gene, while the liver, heart and muscle did not. The results from binding assay were consistent with the fact that Friend MuLV-induced pathogenesis was usually associated with lymphohematopoietic systems, although mRNA expression for mCAT-1 was rather ubiquitous. The discrepancy between F SU/GFP binding and mRNA expression for mCAT-1 in lymphohematopoietic organs of C4W mice would support the receptor interference effect by the Fv-4 (r) gene causing the resistance of C4W mouse to Friend MuLV infection. PMID- 12756623 TI - Development of recombinant coat protein antibody based IC-RT-PCR for detection and discrimination of sugarcane streak mosaic virus isolates from Southern India. AB - Sugarcane streak mosaic virus (SCSMV), causes mosaic disease of sugarcane and is thought to belong to a new undescribed genus in the family Potyviridae. The coat protein (CP) gene from the Andhra Pradesh (AP) isolate of SCSMV (SCSMV-AP) was cloned and expressed in Escherichia coli. The recombinant coat protein was used to raise high quality antiserum. The CP antiserum was used to develop an immunocapture reverse transcription-polymerase chain reaction (IC-RT-PCR) based assay for the detection and discrimination of SCSMV isolates in South India. The sequence of the cloned PCR products encoding 3'untranslated region (UTR) and CP regions of the virus isolates from three different locations in South India viz. Tanuku (Coastal Andhra Pradesh), Coimbatore (Tamil Nadu) and Hospet (Karnataka) was compared with that of SCSMV-AP. The analysis showed that they share 89.4, 89.5 and 90% identity respectively at the nucleotide level. This suggests that the isolates causing mosaic disease of sugarcane in South India are indeed strains of SCSMV. In addition, the sensitivity of the IC-RT-PCR was compared with direct antigen coating-enzyme linked immunosorbent assay (DAC-ELISA) and dot-blot immunobinding assays and was found to be more sensitive and hence could be used to detect the presence of virus in sugarcane breeding, germplasm centres and in quarantine programs. PMID- 12756624 TI - Phylogeny of isolates of Prunus necrotic ringspot virus from the Ilarvirus Ringtest and identification of group-specific features. AB - Isolates of Prunus necrotic ringspot virus (PNRSV) were examined to establish the level of naturally occurring sequence variation in the coat protein (CP) gene and to identify group-specific genome features that may prove valuable for the generation of diagnostic reagents. Phylogenetic analysis of a 452 bp sequence of 68 virus isolates, 20 obtained from the European Union Ilarvirus Ringtest held in October 1998, confirmed the clustering of the isolates into three distinct groups. Although no correlation was found between the sequence and host or geographic origin, there was a general trend for severe isolates to cluster into one group. Group-specific features have been identified for discrimination between virus strains. PMID- 12756625 TI - Molecular evidence that aphid-transmitted Alpinia mosaic virus is a tentative member of the genus Macluravirus. AB - Alpinia mosaic virus (AlpMV), once assigned to the genus Potyvirus, infects primarily plants of the ginger family. To seek molecular evidence for correct classification of this virus, a cDNA clone corresponding to the 3' portion of the AlpMV genome was obtained by reverse transcriptase-PCR and TA cloning. The authenticity of the cDNA clone was confirmed by expression of the coat protein (CP) in E. coli followed by immunoblot analysis. Sequence analysis indicated that, in contrast to its low identity with all the other genera of the family Potyviridae, the deduced amino acid sequence of AlpMV CP was 42.9 - 61.9% identical to members of the genus Macluravirus. Phylogenetic analysis also demonstrated that the AlpMV CP clustered with those of Cardamom mosaic virus and Chinese yam necrotic mosaic virus. These results indicate that AlpMV should be classified as a tentative species within the genus Macluravirus rather than Potyvirus as proposed previously. PMID- 12756626 TI - Molecular characterization of a distinct potyvirus from whitegrass in China. AB - Apotyvirus isolated from perennial whitegrass ( Pennisetum centrasiaticum Tzvel.) in North China was characterized at the molecular level. The 3' terminal nucleotide (nt) sequence of 1669 nt of the viral RNA genome has been determined, which covered the coding region of the C-terminal part of the large nuclear inclusion protein (NIb, RNA polymerase), capsid protein (CP) gene and the 3' nontranslated region (NTR). The CP gene consisted of 909 nt (including the stop codon) encoding 302 amino acid residues, and the 3' NTR was 241 nt in length excluding the polyadenylated tract. Sequence comparison of the amino acids of CPs showed that this virus was most closely related to Sorghum mosaic virus and Maize dwarf mosaic virus with percent identities of 77% to 78% while that of the 3' NTRs suggested that it was most closely related to Zea mosaic virus with identity of 72%. This virus isolate was to some extent closely related to other members of the Sugarcane mosaic virus subgroup of potyviruses for the CP amino acid sequences. Phylogenetic analyses of the sequences indicated that this virus isolate represented a distinct potyvirus, and the name Pennisetum mosaic virus (PenMV) is proposed. PMID- 12756627 TI - Identification of the pocket factors in a picornavirus. AB - Bovine enterovirus (BEV), along with other enteroviruses and the rhinoviruses, has a hydrophobic pocket within structural protein VP1. In the crystal structures of these viruses there is electron density commensurate with a non-protein molecule within the pocket. These molecules, termed pocket factors, have been shown to stabilise the capsid and their removal from the pocket is a necessary prerequisite to uncoating. The pocket factors have been proposed, from the electron densities and uncoating studies, to be short chain fatty acids. In order to identify the pocket factor of BEV, we have grown and purified the virus in an identical manner to that used for the crystal structure determination and have performed a lipophilic extraction. Palmitic acid, C(16:0), was the most abundant accounting for 40.8% by mass of the lipophilic extract (39.3 mol%). Myristic acid C(14:0), was next most abundant at 18.5% by mass (20.0 mol%). In addition, we have identified other fatty acids in smaller proportions. We have therefore shown that BEV contains saturated fatty acid pocket factors of varying chain length. We have also compared the profile of the fatty acyl chain composition of BEV with those for uninfected BHK-21 cell plasma membrane and endoplasmic reticulum extracts. PMID- 12756628 TI - Virus nomenclature; continuing topicality. PMID- 12756629 TI - Pain 5 years after instrumented and non-instrumented posterolateral lumbar spinal fusion. AB - Pain drawings have been used in spine surgery for diagnostic use and psychological evaluation of fusion candidates; they have rarely been used to evaluate pain status after spinal fusion. This study is a 5-year follow-up on a randomised clinical trial assigning patients to posterolateral spinal fusion with or without pedicle screw instrumentation. Patients were mailed a pain drawing and questionnaires including questions regarding work, social status, smoking status, the Dallas Pain Questionnaire (DPQ), and the Low Back Pain Rating Scale (LBPRS). Pain drawings were scored using a visual inspection method and a surface-based point scoring and evaluated for the presence of donor site pain. Pain drawings from 109 patients (87% of the initially included patients), 56 men and 53 women, mean age at follow-up 51 years, were analysed. Fifty-three patients had undergone an instrumented fusion and 56 a non-instrumented fusion. Some presence of low back pain was marked by 79% and leg pain by 69%. Sixty-two percent of the pain drawings were classified as "organic" and 38% as "non-organic". There was no difference between the instrumented and the uninstrumented group. DPQ and LBPRS scores were higher in the non-organic group ( P=0.007). Using the point scoring, no difference between the instrumented and the uninstrumented group was seen. The results of the point scoring were found to correlate with the DPQ and LBPRS scores ( P=0.001). Working patients (39%) had significantly better scores than the rest. Ten percent of the patients had donor site pain. Twenty percent of spinal fusion patients are totally pain free at 5-year follow-up. Ten percent still experience donor site pain. In general, instrumentation does not affect the amount and localisation of pain 5 years after lumbar spinal fusion surgery. The pain drawing seems to be a valuable tool when following spinal fusion patients, but its use as prognostic marker in connection with fusion surgery needs further investigation. PMID- 12756630 TI - Mortality rate and relative strain index in Buenos Aires city. AB - Thermal stress may seriously affect human health to the extent of provoking death in those human groups at high risk such as little children and elderly persons. This research studies the climatology of the relative strain index (RSI) in the city of Buenos Aires (Argentina). It is observed that the conditions at 1400 hours local time on more than 75% of summer days cause discomfort, though only on 25% of days are physiological failure levels reached. The nocturnal RSI values show 75% of nights to be comfortable, bringing relief and allowing physiological recuperation. Comparison between the RSI frequency distributions of different decades by the application of the Kolmogorov-Smirnov test shows a possible urban heat-island effect that increases nocturnal RSI values. The relationship between the RSI and the daily number of interments is also analysed. Because of the large number of different causes of mortality, only about 10% of the variance of the daily number of interments can be explained by the RSI value at 1400 hours local time 1 or 2 days before. Comparison of mortality figures with these RSI values shows deaths to be at a minimum for the interval 0.0-0.1 (the RSI comfort range), slightly increasing for negative values that could represent unexpected Summer cold waves. For values of RSI over 0.2 the increment is more abrupt, indicating an impact of a thermal strain on mortality. The relationship between the joint occurrence of the 1400 hour and 0200 hour (local time) RSI values and mortality shows a clear impact of prolonged exposure to uncomfortable conditions. PMID- 12756631 TI - Increases in cytokine and antimicrobial peptide gene expression in horses by immunomodulation with Propionibacterium acnes. AB - Immunomodulation with Propionibacterium acnes is used for prophylaxis of respiratory disease or in horses suffering from chronic pulmonary inflammation; however, the mechanism for this response is poorly understood. Semiquantitative reverse transcriptase-polymerase chain reaction assays were used to evaluate gene expression of interleukin (IL)-2, IL-10, interferon (IFN)-gamma, and NK-lysin in healthy horses treated with P. acnes. Findings in the study indicated that horses treated with a P. acnes-based immunomodulator exhibited increased IFN-gamma and NK-lysin gene expression in peripheral blood mononuclear cells. These results suggest that part of the immunostimulating properties of a P. acnes-based immunomodulator is derived from enhanced gene expression of the type-1 cytokine IFN-gamma and NK-lysin, an antimicrobial peptide. PMID- 12756633 TI - Evaluation of a modified live virus type-1a bovine viral diarrhea virus vaccine (Singer strain) against a type-2 (strain 890) challenge. AB - Both type-1 and type-2 bovine viral diarrhea virus (BVDV) infections are responsible for major losses in the cattle industry. However, several commercial BVDV vaccines contain only a type-1 strain. A vaccine trial was conducted to evaluate the efficacy of BVDV type-1 (Singer strain; BVDV-1) vaccine for protecting calves challenged with virulent BVDV type-2 (890 strain; BVDV-2). Thirty-eight BVDV-negative calves were randomly allocated to four groups. One group was treated with a modified live virus (MLV) BVDV-1 vaccine by i.m. injection and another group was treated with the same vaccine by s.c. injection. Two groups served as nonvaccinated controls (one i.m. and one s.c.). Twenty-eight days following vaccination, the calves were challenged with BVDV-2 and monitored for 21 days. Clinical scores and body temperatures of vaccinated calves were significantly (P<.05) lower than for controls on several days, and peak differences occurred 8 days after challenge. The control calves had significantly (P<.05) lower leukocyte counts 3 through 8 days after challenge; leukocyte counts for vaccinated animals did not decline significantly from prechallenge levels. There were no differences in protection between the i.m. and s.c. routes of vaccination. The study demonstrated satisfactory cross protection of the BVDV-1 vaccine against BVDV-2 challenge. PMID- 12756632 TI - Evaluation of an ear cleanser for the treatment of infectious otitis externa in dogs. AB - Thirty-one ears (16 dogs) with otitis externa originating from bacterial or yeast infections were enrolled in a study to evaluate the in vivo efficacy of an ear cleanser containing 2.5% lactic acid and 0.1% salicylic acid for the treatment of infectious otitis externa. The affected ears were treated with the ear cleanser twice daily for 2 weeks and evaluated after 1 and 2 weeks of treatment. The ear cleanser was effective in resolution of infection in 67.7% of the ears, and clinical signs of infectious otitis externa were significantly reduced within 2 weeks. PMID- 12756634 TI - Effects of the glycoprotein IIb/IIIa antagonist abciximab on thrombus formation and platelet function in cats with arterial injury. AB - An established model of arterial injury was used to evaluate the ability of the glycoprotein (GP) IIb/IIIa antagonist abciximab to inhibit platelet function and deter thrombus formation in cats. The study was a blinded evaluation, with control cats receiving aspirin and placebo and treatment cats receiving aspirin plus abciximab. The group treated with aspirin plus abciximab had significantly (P<.05) less thrombus formation based on a smaller number and frequency of cyclic flow reductions and reduced luminal thrombus area than cats treated with aspirin plus placebo. Abciximab administered in addition to aspirin also resulted in significant (P<.05) inhibition of platelet function based on mucosal bleeding time and change in bleeding time from baseline. PMID- 12756635 TI - Efficacy of two 65% permethrin spot-on formulations against induced infestations of Ctenocephalides felis (Insecta: Siphonaptera) and Amblyomma americanum (Acari: Ixodidae) on beagles. AB - The efficacy of two formulations of a topically applied 65% permethrin spot-on (Defend Exspot Treatment for Dogs, Schering-Plough Animal Health) was evaluated against experimental infestations of the cat flea Ctenocephalides felis and the lone star tick Amblyomma americanum in dogs. Eighteen dogs were randomly assigned to treatment with 65% permethrin in either diethylene glycol monomethyl ether (DGME; original formulation) or propylene glycol monomethyl ether (PGME) or to be untreated as a control. Treated dogs received either 1 (body weight < 15 kg) or 2 ml (body weight > or =15 kg) of the assigned formulation on Day 0. One hundred unfed, adult C. felis were placed on each dog on Days -6, -1, 4, 11, 18, 25, and 32. Fifty unfed, adult ticks were placed on each dog on Days -1, 3, 9, 16, 23, and 30. Live fleas and ticks were counted and removed on Days 3, 7, 14, 21, and 28. Treatment of dogs with the 65% permethrin in DGME reduced flea numbers by 90.4% to 99.9% from Days 3 through 21 (P < or =.05) and by 48.2% 28 days after treatment. Treatment of dogs with 65% permethrin in PGME reduced flea numbers by 93.7% to 99.7% from Days 3 through 28 and by 78.4% 35 days after treatment (P < or =.05). Treatment with 65% permethrin in DGME reduced tick numbers by 90% or more only on Day 7, whereas treatment with 65% permethrin in PGME reduced the number of live ticks by 90%or more on Days 7 and 14 and approached 90%(87.9%) on Day 21. Efficacy against fleas and ticks for the PGME formulation was significantly better (P < or =.05) than for the DGME formulation on Day 28. Findings in this study indicate that both the DGME and PGME formulations of 65% permethrin performed well in reducing numbers of live C. felis and A. americanum on laboratory beagles; however, the PGME formulation was effective approximately 1 to 2 weeks longer than the DGME formulation. PMID- 12756636 TI - Use of client-specific outcome measures to assess treatment effects in geriatric, arthritic dogs: controlled clinical evaluation of a nutraceutical. AB - A questionnaire method was designed for dog owners to monitor the orthopedic disabilities of their pets for evaluation of a nutraceutical with joint health claims. Fifty large-breed dogs, 7 to 12 years of age, presenting with signs of osteoarthritis, were randomly allocated to placebo and active treatment groups. Degree of disability was assessed by physical examination, a standard questionnaire on daily activities, and a case-specific questionnaire that monitored specific impairments of each dog. The test product was a special milk protein concentrate (SMPC) from hyperimmunized cows, previously shown to express antiinflammatory and antiarthritic activity in humans. After a 1-week run-in period of dosing with placebo, each dog was randomly assigned to a treatment and given gelatin capsules containing either SMPC or a placebo twice daily for 8 weeks. Overall improvement was noted in 68% and 35% of the SMPC and placebo groups, respectively. Significant (P <.05) improvement in mean standardized and patient- specific questionnaire scores and in owner global assessments was detected in the SMPC group but not in the placebo group. Compared with the placebo group, the treatment response was significantly better in the SMPC group with regard to case-specific scores (P <.001) and owner global assessments (P =.004). The product was well tolerated and serum chemistry findings remained within normal limits. PMID- 12756637 TI - Serologic prevalence of Dirofilaria immitis, Ehrlichia canis, and Borrelia burgdorferi infections in Brazil. AB - Dogs infected with Dirofilaria immitis, Ehrlichia canis, or Borrelia burgdorferi may show nonspecific clinical signs or may be asymptomatic. In Brazil, E. canis and D. immitis infections are frequently diagnosed based on the presence of classical signs; however, serologic tests are seldom performed to confirm the presence of infection. To estimate the seroprevalence of these three canine diseases in Brazil, 2,553 dogs presented at veterinary practices for various tests, routine treatments, or examinations were evaluated by an in-office commercial ELISA test kit (SNAP 3Dx, IDEXX Laboratories). Each dog was examined by the veterinarian, and a whole-blood sample was collected and immediately tested for the simultaneous detection of B. burgdorferi and E. canis antibodies and D. immitis antigen. D. immitis infection was detected in 51 dogs (2.0%) and E. canis antibodies were present in 505 dogs 19.8%). Only one dog tested positive for B. burgdorferi antibodies. PMID- 12756638 TI - Preclinical tolerance and pharmacokinetic assessment of MU-Gold, a novel chemotherapeutic agent, in laboratory dogs. AB - MU-Gold, tetrakis (trishydroxymethyl) phosphine gold(I) chloride, a novel gold compound, has cytotoxic effects against human androgen-dependent and -independent prostatic, gastric, and colonic carcinoma in cell culture and against malignant lymphoma in rodent models. A pilot study was conducted to evaluate the tolerance and pharmacokinetic properties of MU-Gold in normal dogs in anticipation of clinical trials in cancer-bearing dogs. MU-Gold (10 mg/kg) was administered by i.v. injection to three purpose-bred dogs. Serum was collected from all dogs for measurement of gold levels via atomic absorption spectrometry. In addition, complete blood counts and biochemical profiles were monitored for Dogs 2 and 3 every 7 days for 30 days. A two-compartment i.v. bolus model with first-order kinetics, mean elimination half-life of approximately 40 hours, and mean volume of distribution of 0.6 L/kg was established. Serum gold concentrations ranging from 10 to 50 mcg/ml were sustained for 2 to 3 days with no clinically significant toxicities observed. Based on in vitro results in earlier studies and preliminary pharmacokinetic data collected in the present study, Phase I clinical trials should be conducted to define the optimal dosage, dose-limiting toxicities, and other characteristics of MU-Gold that will be used to design Phase II clinical trials. PMID- 12756639 TI - A comprehensive review of ceftiofur sodium and hydrochloride formulations for treatment of acute bovine foot rot. AB - Seven well-controlled studies conducted under multiple management conditions demonstrated that ceftiofur, a late-generation veterinary parenteral cephalosporin, is effective for the treatment of bovine foot rot in beef and dairy cattle. Two preliminary dosage titration studies using a challenge model compared the efficacy of ceftiofur (1.1 mg or 2.2 mg ceftiofur equivalents [CE]/kg administered once daily for 3 days) with placebo. One preliminary clinical study evaluated the efficacy of ceftiofur sodium (1.0 mg CE/kg once daily for 3 days) in lactating dairy cows. Two clinical trials evaluated the efficacy of ceftiofur sodium versus placebo for naturally occurring foot rot, and two trials compared the efficacy of ceftiofur sodium or hydrochloride (1.0 mg CE/kg) with oxytetracycline (6.6 or 10 mg/kg), each administered once daily for 3 days, for treatment of acute foot rot in beef cattle. All trials demonstrated the efficacy of ceftiofur for treatment of acute bovine foot rot. Ceftiofur and oxytetracycline were comparable in efficacy, with ceftiofur having excellent injection-site tolerance and short or no milk discard or preslaughter withdrawal. PMID- 12756640 TI - Effects of an ethyl lactate shampoo in conjunction with a systemic antibiotic in the treatment of canine superficial bacterial pyoderma in an open-label, nonplacebo-controlled study. AB - An open-label, nonplacebo-controlled study was designed to compare systemic cephalexin therapy versus systemic cephalexin and ethyl lactate shampoo therapy in the treatment of canine superficial bacterial pyoderma. Twenty client-owned dogs diagnosed with generalized superficial bacterial pyoderma (SP) were alternately assigned to oral treatment with cephalexin (25 to 30 mg/kg every 12 hours) or treatment with cephalexin (as for Group 1) and twice-weekly shampooing with a 10% ethyl lactate shampoo, which was left in contact with the dog's skin for 10 minutes. On Days 14 and 28, skin lesion severity scores, assessed by the investigators, were significantly (P <.01) lower for the group treated with cephalexin and shampoo than for the group treated with cephalexin only. On Day 14, dog owners gave better scores to dogs treated with cephalexin and shampoo for haircoat appearance and body odor than for dogs treated only with cephalexin. Clinical and cytologic resolution of SP occurred significantly (P <.02) sooner in the cephalexin/shampoo group (29.4 days) than in the cephalexin only group (37.8 days). PMID- 12756642 TI - Positron emission tomography and single photon emission computed tomography in epilepsy care. AB - Radiopharmaceutical brain imaging is clinically applied in planning resective epilepsy surgery. Cerebral sites of seizure generation-propagation are highly associated with regions of hyperperfusion during seizures, and with glucose hypometabolism interictally. For surgical planning in epilepsy, the functional imaging modalities currently established are ictal single photon emission computed tomography (SPECT) with [(99m)Tc]technetium-hexamethylpropyleneamine oxime (HMPAO) or with [(99m)Tc]technetium-ethylene cysteine dimer (ECD), and interictal positron emission tomography (PET) with 2-[(18)F]fluoro-2-deoxyglucose (FDG). Ictal SPECT and interictal FDG PET can be used in presurgical epilepsy evaluations to reliably: (1) determine the side of anterior temporal lobectomy, and in children the area of multilobar resection, without intracranial electroencephalographic recording of seizures; (2) select high-probability sites of intracranial electrode placement for recording ictal onsets; and, (3) determine the prognosis for complete seizure control following anterior temporal lobe resection. Coregistration of a patient's structural (magnetic resonance) and functional images, and statistical comparison of a patient's data with a normal data set, can increase the sensitivity and specificity of these SPECT and PET applications to the presurgical evaluation. PMID- 12756643 TI - Imaging studies in movement disorders. AB - Imaging presynaptic dopaminergic markers provides key insights into the pathophysiology of Parkinson's Disease (PD) and is becoming an important endpoint in clinical trials of potential disease-modifying therapies for PD. The further development of this area includes work to optimize targets for accurate and reliable measurement of disease progression. Ultimately, it may be possible to elaborate these markers to fine-tune our understanding of those patients who might be enrolled in a trial. For example, PD patients may be characterized as slow vs. fast progressors based on imaging measures, providing the opportunity to optimize the trial recruitment to demonstrate the greatest impact in Phase 2 evaluations of neuroprotective agents. Further, while dopamine degeneration is a crucial feature of PD, it is clear that there is widespread degeneration in the brain in PD and that many clinical manifestations of PD are likely not due to dopamine deficiency. It is reasonable to imagine that the characterization of additional targets outside the dopamine system could aid in both the molecular basis for disease characterization and ultimately optimization of therapeutics. PMID- 12756644 TI - Positron emission tomography and single-photon emission computed tomography in substance abuse research. AB - Many advances in the conceptualization of addiction as a disease of the brain have come from the application of imaging technologies directly in the human drug abuser. New knowledge has been driven by advances in radiotracer design and chemistry and positron emission tomography (PET) and single-photon emission computed tomography (SPECT) instrumentation and the integration of these scientific tools with the tools of biochemistry, pharmacology, and medicine. This topic cuts across the medical specialties of neurology, psychiatry, oncology, and cardiology because of the high medical, social, and economic toll that drugs of abuse, including the legal drugs, cigarettes and alcohol, take on society. This article highlights recent advances in the use of PET and SPECT imaging to measure the pharmacokinetic and pharmacodynamic effects of drugs of abuse on the human brain. PMID- 12756645 TI - Applications of positron emission tomography in psychiatry. AB - Blood flow, metabolism, and structural imaging studies suggest altered neural circuits in major psychiatric disorders including mood disorders, schizophrenia, and obsessive compulsive disorder. Neuroreceptor mapping studies have identified serotonergic abnormalities in mood disorders and dopaminergic abnormalities in schizophrenia. Further imaging applications have involved development of new positron emission tomography (PET) tracers that may identify abnormalities in peptide neurotransmitter systems such as corticotrophin releasing factor or substance P. Finally, PET can play an important role in quantifying the relationship between receptor occupancy, drug blood levels, oral dose and therapeutic outcome. In that way PET scanning can contribute to both therapeutics and to drug development by more rapid identification of the likely therapeutic dose range compared with conventional parallel group dose comparisons or dose ranging studies. PMID- 12756646 TI - Neuroimaging in patients with head injury. AB - Head trauma affects thousands of people every year. Neuroimaging techniques provide some of the most important diagnostic, prognostic, and pathophysiological information in the management of brain injury. Anatomical imaging modalities can help assess intracranial hemorrhage, fractures, and other structural lesions. Functional imaging has been shown to be helpful in assessing the areas of the brain affected by the trauma as well as determining long term prognosis and rehabilitation potential. This article will review the current uses of neuroimaging techniques in head trauma and delineate future applications. PMID- 12756647 TI - Imaging gliomas with positron emission tomography and single-photon emission computed tomography. AB - Over the last two decades the large volume of research involving various brain tracers has shed invaluable light on the pathophysiology of cerebral neoplasms. Yet the question remains as to how best to incorporate this newly acquired insight into the clinical context. Thallium is the most studied radiotracer with the longest track record. Many, but not all studies, show a relationship between (201)Tl uptake and tumor grade. Due to the overlap between tumor uptake and histologic grades, (201)Tl cannot be used as the sole noninvasive diagnostic or prognostic tool in brain tumor patients. However, it may help differentiating a high-grade tumor recurrence from radiation necrosis. MIBI is theoretically a better imaging agent than (201)Tl but it has not convincingly been shown to differentiate tumors according to grade. MDR-1 gene expression as demonstrated by MIBI does not correlate with chemoresistance in high grade gliomas. Currently, MIBI's clinical role in brain tumor imaging has yet to be defined. IMT, a radio labeled amino acid analog, may be useful for identifying postoperative tumor recurrence and, in this application, appears to be a cheaper, more widely available tool than positron emission tomography (PET). However, its ability to accurately identify tumor grade is limited. 18 F-2-Fluoro-2-deoxy-d-glucose (FDG) PET predicts tumor grade, and the metabolic activity of brain tumors has a prognostic significance. Whether FDG uptake has an independent prognostic value above that of histology remains debated. FDG-PET is effective in differentiating recurrent tumor from radiation necrosis for high-grade tumors, but has limited value in defining the extent of tumor involvement and recurrence of low-grade lesions. Amino-acid tracers, such as MET, perform better for this purpose and thus play a complementary role to FDG. Given the poor prognosis of patients with gliomas, particularly with high-grade lesions, the overall clinical utility of single photon emission computed tomography (SPECT) and PET in characterizing recurrent lesions remains dependent on the availability of effective treatments. These tools are thus mostly suited to the evaluation of treatment response in experimental protocols designed to improve the patients' outcome. PMID- 12756648 TI - [Islet of Langerhans transplantation in type I diabetes. Regulation of neovascularization in the transplant is a key question]. PMID- 12756649 TI - [Interviews with sexually abused children have often wrong aims. What are we prepared to listen to?]. PMID- 12756650 TI - [Severe vascular dysfunction shown in transplanted islets]. AB - Despite recent advances in clinical islet transplantation, a surprisingly large number of islets (approximately 1 million) are still required to obtain insulin independence in type 1 diabetes. The reasons for this are obscure and likely multifactorial. One explanation may be disturbances in engraftment of the transplanted islets, i.e. the adaptation of the islet transplant to its new surroundings with regard to e.g. revascularization and blood perfusion. Endogenous islets have a dense glomerular-like angioarchitecture. Transplantation of isolated islets causes a disruption of their vascular connections, making the islets dependent on the formation of new blood vessels for optimal function. Evidence from experimental islet transplantation indicates an insufficient revascularization of transplanted islets with subsequent chronically decreased blood perfusion and oxygen tension, which has metabolic consequences within the tissue. PMID- 12756651 TI - [Time to institute more active follow up after curative surgery for colorectal cancer? Wide variation of follow up and diagnostic routines]. AB - There are still no large controlled studies that have proved any survival benefit with intensive follow-up after curative surgery for colorectal cancer. There is a wide variation in follow-up. Three meta-analysis have shown that intensive follow up can improve survival, the most recent based on five previously published small controlled trials that compared intensive with control follow-up. Since the randomised studies were initiated, there have been a strong development of rectal cancer surgery and surgery for livermetastasis as well as for adjuvant and palliative oncologic treatments. This development gives support to a more active attitude towards intensive regular follow-up. An active follow-up should be selective and only include patients who tolerate oncologic or surgical treatment. Diagnostic tests to find intraluminal recurrence are ineffective. Large multicenter studies are still warranted. PMID- 12756652 TI - [Injuries and mortality in motorcycle and moped accidents in Sweden 1987-1994. Advanced age and male sex are risk factors of fatal moped and motorcycle accidents]. AB - Data from the Swedish Hospital Discharge Registry were analysed, years 1987-1994 inclusive. The registry includes all living patients admitted to Swedish hospitals. Patients admitted after moped or motor-cycle accidents were studied. A total of 4,716 mopedists and 8,927 motor-cycle riders were admitted in the given time frame. The median age of mopedists was 16 and that of motor-cycle riders 22 years. 85% of mopedists were male, while 95% of motor-cycle drivers and 55% of motor-cycle passengers were male. The injury distribution in both groups was quite similar with fractures and cerebral concussion dominating, although fractures of the arm and vertebrae were considerably more common in motor-cycle riders. The incidence of hospital admission after accidents did not change over time in mopedists, but decreased in motorcyclists. Accidents involving both types of vehicles occurred more commonly in the countryside. High age and male sex was associated with increased mortality in mopedists, while high age and being a passenger was associated with higher death rates in admitted motor-cyclists. PMID- 12756653 TI - [A study of primary health care in 15 different countries. Examination and treatment of patients with heart failure should be better]. PMID- 12756654 TI - [Serendipity--insight of unexpected connections]. PMID- 12756655 TI - [Primus Mortimer Pettersson--one of our leading naivists. He created a world of beauty, tranquility and alarm]. PMID- 12756656 TI - [Medicalization of poverty--illness seen from a primary health care's point of view]. PMID- 12756657 TI - [The 3G net and "pensioners' computers" can help to solve problems in health care]. PMID- 12756659 TI - [Proposal for a new structure of the specialist education should not be implemented]. PMID- 12756658 TI - [A new interesting study on coxib]. PMID- 12756660 TI - [Needle-exchange programs can not be defended]. PMID- 12756661 TI - [A staggering blow if I am not allowed to practice my surgery]. PMID- 12756662 TI - [Cobalamin and folate from the cradle to the grave!]. PMID- 12756663 TI - [About the content of the freedom of speech--psychiatric research should tolerate critical scrutiny, too]. PMID- 12756664 TI - New ads promote "sip all day, get decay" massage. PMID- 12756665 TI - Legalize denturism? PMID- 12756666 TI - Our fair share. PMID- 12756667 TI - A new way to look at heart disease? PMID- 12756668 TI - Reporting intoxicated patients; good samaritan immunity. PMID- 12756669 TI - Successfully working with people you don't like. PMID- 12756670 TI - A safety checklist for your office. PMID- 12756671 TI - Are your patients depressed? Implications for dental practice. AB - Depressive disorders traditionally reside outside the realm of customary dental practice. Nonetheless, one in every five patients who visits a dentist experiences clinically significant symptoms of depression. The clinical implications of this are substantial. Depression is associated with diminished salivary flow and the complaint of dryness of mouth. It is associated with a diminished and distorted taste sensation, and a higher oral lactobacillus count. Depression is a risk factor for the development of dental caries, periodontal disease, and the erosive variant of oral lichen planus. Antidepressant medications can produce xerostomia, dysgeusia and bruxism. Depressive illness is a legitimate medical condition, with recognizable signs and symptoms, definable pathophysiology, and a significant response to treatment. Unfortunately, despite the availability of effective therapeutic measures, the majority of patients remain untreated. Routine dental checkup visits provide an opportunity for screening. PMID- 12756672 TI - Dr. Martin Tuck, Chair of the Michigan Board of Dentistry. Interview by Michael G. Maihofer. PMID- 12756673 TI - Quiz #13. Oral pathology quiz: chronic ulcer under a denture. Infectious, traumatic and idiopathic disease processes. PMID- 12756674 TI - Alternative products for cancer: a benefit or risk to the patient? PMID- 12756675 TI - The nursing shortage: myth or reality. PMID- 12756676 TI - Syncop.aging. n. The art and science of syncope in the aged. PMID- 12756677 TI - C-reactive protein. Should it be considered a coronary risk factor? AB - C-reactive protein (CRP) is an acute phase reactant which is not associated with coronary atherosclerosis in many studies. However, it has been demonstrated in many, but not all, studies to predict cardiovascular events. Increased CRP levels may reflect tissue damage and inflammation not only in the arteries, but anywhere in the body. Elevated CRP levels may be induced by metabolic, infective, immunologic, or other processes. Increased CRP levels are probably an indirect marker of any increased cytokine response to inflammatory stimuli that are critical for atherosclerotic plaque progression and rupture. A large-scale prospective trial is needed to investigate whether reduction of elevated CRP will reduce cardiovascular events. PMID- 12756678 TI - Syncope. Identifying cardiac causes in older patients. AB - Syncope is a transient loss of consciousness and postural tone, with spontaneous recovery, and may represent an episode of aborted cardiac arrest. The final common pathway for loss of consciousness from non-neurologic causes is hypoperfusion of the reticular activating system. The etiology of syncope in older patients frequently includes a myriad of causes (e.g., orthostatic hypotension, medication side effects, sick sinus syndrome, atrioventricular block, carotid hypersensitivity, ventricular tachycardia, and aortic stenosis). Initial evaluation must include a detailed history, complete physical examination, and 12-lead ECG, which provides a framework as to the probable cause of syncope in approximately 40% of patients. Echocardiography, electrophysiology testing, and head-upright tilt table testing will confirm the diagnosis in most of the remainder. Approximately 25% of patients will have no identifiable cause of syncope, but have a better prognosis than patients with a confirmed cardiac cause of syncope. Treatment is directed at the underlying cause. PMID- 12756679 TI - Statins. Evidence of effectiveness in older patients. AB - As the population ages, increasing numbers of older adults are becoming candidates for lipid-lowering therapy with HMG CoA reductase inhibitors (statins) and lifestyle modification. Available evidence shows that statins reduce cardiovascular events and invasive revascularization in older adults. Statins have not only been shown to be safe and effective for lowering LDL cholesterol, but appear to have ancillary pleiotropic effects that are beneficial in other conditions to which older adults are prone. Despite these benefits, older adults are currently undertreated for this highly treatable cardiovascular risk factor. PMID- 12756680 TI - Rash on the lower back. History of low-back pain leads to the source of the problem. PMID- 12756681 TI - Anxiety in older adults. Assessment and management of three common presentations. PMID- 12756682 TI - Expired research. PMID- 12756683 TI - Intrusion: the central problem for family health promotion among children and single mothers after leaving an abusive partner. AB - Like other single-parent families, those consisting of mothers and their children who leave abusive partners/fathers are broadly viewed a deficient, high-risk structures in which children are susceptible to multiple problems. The mechanisms of strength and vulnerability in these families are poorly understood, and, consequently, their health promotion processes remain virtually unexplored. In a feminist grounded theory study of health promotion processes of single-parent families after leaving abusive partners/fathers, the authors discovered intrusion to be the basic social problem as families strive to promote health in the aftermath of abuse. The authors discuss the complex nature of intrusion, demonstrating how health is socially determined, and the challenges of health promotion in terms of the issues and dilemmas faced by study families and consider implications for health promotion knowledge and practice. PMID- 12756684 TI - Home health care nurses' perceptions of physician-nurse relationships. AB - This work is a sociological analysis of the physician-nurse relationship in home health care. Data include observation, interview, and survey sources. Results confirm that, largely because of the physician's distance both physically and symbolically, home care nurses take on a more primary care role with patients than do nurses in hospitals. There are fewer instances of conflict between physician and nurse in this setting, as doctors yield more responsibility to nurses than they may in hospitals. In the end, though nurses claim higher levels of autonomy, patients appear to be the true beneficiaries of the in situ shift in nursing responsibility. PMID- 12756685 TI - Factors underlying anxiety in HIV testing: risk perceptions, stigma, and the patient-provider power dynamic. AB - Client anxiety is often associated with diagnostic testing. In this study, the authors used a grounded theory approach to examine the situational and social factors underlying anxiety associated with HIV testing, analyzing transcripts from semistructured interviews with 39 HIV test recipients in Ontario, Canada (selected based on HIV serostatus, risk experience, geographic region, gender, and number of HIV tests), then integrating emergent themes with existing research literature. Analysis revealed four themes: perceptions of risk and responsibility for health, stigma associated with HIV, the patient-provider power dynamic, and techniques used by test recipients to enhance control in their interactions with providers. Service implications include modifications to information provision during the test session, attention to privacy and anonymity, and sensitivity to patient-provider interactions. PMID- 12756686 TI - Living the golden rule: reciprocal exchanges among African Americans with cancer. AB - Giving is receiving, and receiving is giving. This is the key finding from interviews conducted with 28 African American women and men with cancer who were active participants in dynamic relationships characterized by both giving and receiving. These participants engaged in reciprocal relationships varying in the number of persons involved, types of resources exchanged, and timing of exchange. Findings suggest the need to reconceptualize social support as caregiving and caregiving as social support. This study also points to the need to redesign intervention studies to be more inclusive of components that allow the elderly in illness-related situations to maintain their status as givers in their social networks. PMID- 12756687 TI - Reconstructing a meaning of pain: older Korean American women's experiences with the pain of osteoarthritis. AB - The purpose of this grounded theory study was to gain a deeper understanding of older Korean American women's experiences of chronic osteoarthritic pain. The data included a set of 3 interviews with 7 women over age 60, field notes, observations, memos, and the literature about the chronic pain of osteoarthritis. Through constant comparative analysis and coding typical of grounded theory, a five-stage process emerged inductively from the data. The core variable, Reconstructing a Meaning of Pain, included the concepts of the process in the grounded theory through which the women learned to manage and tolerate the pain. During this process, the women came to perceive their pain as a component of aging rather than as a symptom of disease. PMID- 12756688 TI - Legal, financial, and ethical ambiguities for Mexican American families: caring for children with chronic conditions. AB - The author reports findings from a study about experiences of 17 Mexican American families caring for children with serious chronic conditions. Legal, financial, and ethical ambiguities arose when parents' desire to provide necessary care for their children and providers' professional commitment to offer this care conflicted with United States laws, including welfare reform initiatives, requiring providers to determine eligibility before providing care to immigrants and to report undocumented care seekers to authorities. Families frequently felt intimidated because health care systems are complex, and legal residency status often varied among family members. Findings imply that official policy and education of family members should aim to assure that children with chronic conditions receive needed services without relying on providers to enforce immigration laws. PMID- 12756689 TI - Understandings of health and its determinants among clients and providers at an urban community health center. AB - The authors' aim was to explore clients' and health care providers' perceptions of health and its determinants in preparation for the development of a screening tool to assess client health determinants at a community health center through template and editing analyses of interviews with 6 health care providers and 7 clients. Participants defined health as a multidimensional state, with common themes including the ability to cope and to function according to expectations. They identified multiple interrelated factors that affect health. The findings support existing health determinant frameworks but provide greater detail about specific determinants within broad categories presented in these frameworks. The results create a foundation for the development of a screening tool to assess client health determinants. PMID- 12756690 TI - Overcoming ambivalence: the challenges of exploring socially charged issues. AB - In this article, the author considers the challenges associated with the investigation of sensitive and socially charged issues. Drawing on a qualitative study of older women's body image and embodied experience, she discusses how societal ambivalence toward older women's bodies and appearances shapes and constrains the establishment of rapport between a young researcher and the study participants. Exploring the internalized anxiety expressed by the women, she investigates how life history narratives, multiple interviews, photographs, location of the interview, self-disclosure, information sharing, provision of assistance, student and stranger statuses, collaborative interviewing, and impression management combine to enhance rapport and ease the women's sense of discomfort. PMID- 12756691 TI - [Dieting with (un)known hormones]. PMID- 12756692 TI - [Grades, tests, quota system or drawing of lots for medical school applications? When there are four applications for every single post something must be done]. PMID- 12756693 TI - [Dieting preparations caused difficult-to-interpret thyroid gland disorders. Warning for "Eat&Lose"]. PMID- 12756694 TI - [Home care for elderly cancer patients. More intensive follow-up and home services reduce the need of specialist care]. AB - The effects of intensified primary health care were examined in the "Support-Care Rehabilitation" project conducted in Uppsala county 1993-1997. Intensified primary health care was one part of an individual support intervention and comprised extended information about patients from the specialist clinics, and education and supervision in cancer care for GPs and home care nurses. The aim was to improve the ability of general practitioners and home care nurses to monitor and support cancer patients. A total of 485 newly diagnosed cancer patients were randomized to intensified primary health care or to a control group (standard care). The intervention group showed a marked increase in follow-up contacts. About 90% of intensified primary health care patients reported such contacts, compared to 26% of control patients. The number of days of hospitalization at the specialist clinics for elderly patients (> or = 70 yr.) randomized to the intervention group were 393 less than for elderly controls three months after diagnosis. The conclusion is that intensified primary health care constitutes a cost-effective strategy for enhancing co-operation between home care, primary health care and specialist clinics. PMID- 12756695 TI - [West Nile fever may even be spread to Sweden]. AB - West Nile Virus (WNV) is a flavivirus, which was first isolated in Uganda 1937. This virus has attracted attention in the past years. WNV is one of the worlds most widespread flaviviruses and has caused recent outbreaks among humans and animals. West Nile Fever (WNF) is a mosquito borne zoonotic disease. The last decade the epidemiological pattern of the virus and the severity of the outbreaks have changed. This article reviews the epidemiological changes and current situation of WNF, presents a case report and evaluates the risk for a possible outbreak in Sweden. PMID- 12756696 TI - [A case report: anomaly of the fourth branchial pouch with recurring cervical abscesses. Cauterization with trichloroacetic acid closed the fistula opening and cured the patient]. AB - A fourth branchial pouch sinus is a rare congenital anomaly, which in a 13-year old girl presented clinically as recurrent deep cervical abscesses. The location of the majority of these anomalies is the left side of the neck (90%). Radiological and endoscopic investigations verified the diagnosis. The internal orifice located at the apex of the pyriform sinus could facilitate contamination by infectious pharyngeal secretions and lead to abscess recurrence. Traditionally, the recommended treatment is radical surgery. It can, however, be technically difficult to excise the whole fistula tract. In this patient we used a non-invasive treatment modality; chemocauterization with 40% trichloroacetic acid (TCA). After three treatments the fistula was closed. To date (month no. 15) there has been no abscess recurrence. TCA chemocauterization seems to be a safe first-line treatment for patients with pyriform sinus fistulas. PMID- 12756697 TI - [Diaries and letters indicate that Franz Kafka suffered from Horton headache]. PMID- 12756698 TI - [Injudicious declarations about the benefits of alcohol!]. PMID- 12756699 TI - [Nabumetone is an alternative]. PMID- 12756700 TI - [Interdisciplinary society-oriented analysis is required to understand the "ill health paradox"]. PMID- 12756701 TI - [Sexual experiences in connection with sexual abuse can delay the disclosure]. PMID- 12756702 TI - [Human feelings are exceptional when efficiency measures in health care services take over]. PMID- 12756703 TI - [Dermatological views on orthopedic metal implants]. PMID- 12756705 TI - [Joint policy is the guarantee for the future of private practitioners in Stockholm]. PMID- 12756704 TI - [The issue of organ donation is seriously taken by the intensive care personnel]. PMID- 12756706 TI - [From fire to ashes--and then?]. PMID- 12756707 TI - [Nexium and Losec are equally good]. PMID- 12756708 TI - [Waiting lists for surgery are expensive--depending on how we calculate. No calculation method is without controversies]. PMID- 12756709 TI - [Severe acute respiratory syndrome (SARS). Local infection--global panic]. PMID- 12756710 TI - [Four years on a waiting lists for surgery--an expensive option. Millions in lost production while waiting for an orthopedic intervention]. AB - This study was performed in order to determine the cost to society (in terms of loss of production) of having patients on paid sick-leave while on a waiting list for elective orthopedic surgery. All patients on surgical waiting lists receiving sick-leave benefits for the same diagnosis as for the planned procedure, specifically for lumbar disc herniation, lumbar spinal stenosis, and certain knee and shoulder diagnoses (not including arthritis), were identified at two large Swedish orthopedic clinics. These diagnoses were chosen since there is evidence that surgery can reduce pain and disability and also improve work ability. The number of days on sick-leave was determined individually as was each subject's reimbursement from universal health insurance. These benefits were treated as equal to the production losses caused by their inability to work according to the so-called Human Capital Method. 159 patients on the waiting lists were on sick leave. The average waiting time varied between one and two years for the diagnoses included. Forty-four of the patients were granted temporary or permanent disability pensions while awaiting surgery. The costs for paid sick leave together with future costs for those granted permanent disability pensions were almost 90 million SEK (almost 90 million USD). This amount corresponded to the cost of more than 2000 disc operations or more than 1000 total hip replacements. Instead of being spent on sick-leave this money ought to be used to shorten the waiting time for surgery. PMID- 12756711 TI - [Uneven quality of referrals for lower urinary tract symptoms in men]. AB - Referrals (N = 126) for LUTS (lower urinary tract symptoms) in men over 40 years of age were scrutinized. Descriptions regarding symptom duration, type and intensity were unsatisfactory in 25% of the forms. Not a single work-up prior to referral was completed in accordance with the established protocol. One quarter of the men had previously been referred for LUTS. PMID- 12756712 TI - [Few injuries among Swedish telemark skiers, but equipment requires careful consideration]. AB - A study of the incidence and predisposing factors for telemark injuries was carried out in a population of 504 active Swedish skiers, who reported 17,383 skiing days, during 2 seasons in northern Sweden. This is on an average 17 skiing days per person and season. The average age was 33 years. 90 skiers reported 113 injuries in 103 accidents, thus they reported 6.5 injuries per 1000 skiing days. The most common injury location was the knee, followed by, in decreasing order, trunk, thumb and ankle injuries. Skiers who used binding lifters had an increased risk, while more rigid plastic ski boots and increased skiing skill had a preventing effect. The releasable bindings need refinement to fulfil the desired effect. PMID- 12756713 TI - [Scrutiny of drug marketing by drug industry]. PMID- 12756714 TI - [Nexium is not better than Losec]. PMID- 12756715 TI - [Winter vomiting disease in the county of Kronoberg 2002-2003: more serious sequelae than expected]. PMID- 12756716 TI - [The National Board of Health and Welfare and the National Social Insurance Board: Questions and answers concerning ICD codes on formularies for sickness certification]. PMID- 12756717 TI - [Sick listing--who is to blame, Bernhard Grewin?]. PMID- 12756718 TI - [Smallpox and HIV--a final reply]. PMID- 12756719 TI - Evolution of pathogenic Yersinia, some lights in the dark. PMID- 12756721 TI - Tracing acquisitions and losses in Yersinia genomes. PMID- 12756720 TI - DNA adenine methylation. PMID- 12756722 TI - Subtractive hybridization uncovers novel pathogenicity-associated loci in Yersinia enterocolitica. PMID- 12756723 TI - Identification of genes involved in Yersinia pestis virulence by signature-tagged mutagenesis. PMID- 12756724 TI - Characterization of two conjugative Yersinia plasmids mobilizing pYV. PMID- 12756725 TI - Signature-tagged mutagenesis of Yersinia pestis. PMID- 12756726 TI - Cobalamin synthesis in Yersinia enterocolitica 8081. Functional aspects of a putative metabolic island. PMID- 12756727 TI - Construction of a Yersinia pestis microarray. PMID- 12756728 TI - A conjugal type IV transfer system in Yersinia enterocolitica strains. PMID- 12756729 TI - Transmission factors: Yersinia pestis genes required to infect the flea vector of plague. PMID- 12756730 TI - Rho-GTP binding proteins in Yersinia target cell interaction. PMID- 12756731 TI - A technique of intradermal injection of Yersinia to study Y. pestis physiopathology. PMID- 12756732 TI - YopT is a cysteine protease cleaving Rho family GTPases. PMID- 12756733 TI - Structural studies of Yersinia adhesin YadA. PMID- 12756734 TI - Yersinia pseudotuberculosis harbors a type IV pilus gene cluster that contributes to pathogenicity. PMID- 12756735 TI - Salicylanilides are potent inhibitors of type III secretion in Yersinia. PMID- 12756736 TI - Mapping of possible laminin binding sites of Y. pestis plasminogen activator (Pla) via phage display. AB - We tried to determine amino acid motifs of Y. pestis plasminogen activator (Pla) involved in laminin binding. We selected heptamer peptides using a random phage library which was tested against immobilised laminin. Two sequences seemed to inhibit Pla mediated laminin binding of E. coli and exhibited a strong laminin binding capacity revealing a competition with Pla for the same laminin binding site. The motifs are also involved in plasminogen activation because they caused fibrinolysis on fibrin films. The patterns were localised outside the putative surface-exposed loops (Kukkonen et al., 2001). PMID- 12756737 TI - The fish pathogen Yersinia ruckeri possesses a TTS system. PMID- 12756738 TI - Characterisation of the type III secretion protein YscU in Yersinia pseudotuberculosis. YscU cleavage--dispensable for TTSS but essential for survival. PMID- 12756739 TI - Mutagenesis elucidates the assembly pathway and structure of Yersinia pestis F1 polymer. PMID- 12756740 TI - Characterization of infections with wild and mutant Yersinia pseudotuberculosis strains in rabbit oral model. PMID- 12756741 TI - Identification of Yersinia pestis pigment receptor. PMID- 12756742 TI - Yersinia enterocolitica biotype 1A: not as harmless as you think. PMID- 12756743 TI - Pestoides F, a Yersinia pestis strain lacking plasminogen activator, is virulent by the aerosol route. PMID- 12756744 TI - Impact of the Yersinia pseudotuberculosis-derived mitogen (YPM) on the murine immune system. PMID- 12756745 TI - Role of T cells and gamma interferon in Yersinia pseudotuberculosis-derived mitogen (YPM)-induced toxicity in mice. PMID- 12756746 TI - Yersinia pestis Pla has multiple virulence-associated functions. PMID- 12756747 TI - Polyclonal B-cell activation in mice infected by intragastric route with Yersinia enterocolitica O:8. PMID- 12756748 TI - Polyclonal activation as a consequence of infection of mice with Yersinia enterocolitica O:3 isolated from patients with or without arthritis. PMID- 12756749 TI - The response of murine macrophages to infection with Yersinia pestis as revealed by DNA microarray analysis. AB - Macrophages play a crucial role in recognition and phagocytosis of pathogens and in the induction of response, immunity and immunopathology. A key strategy employed by numerous pathogens such as Yersinia pestis is to circumvent the immune response of the host via actively down-regulating the activation of macrophages. The study on host-pathogen interaction and gene expression is imperative for the development of alternative therapeutics. We have combined Suppression Subtractive Hybridisation (SSH), Microarray techniques, Northern blot analysis and quantitative reverse transcription coupled PCR (RT-PCR) to gain a view of differential host gene expression in response to Y. pestis-26 degrees C infection. In our study, a total of 22 different genes were identified as up regulated in response to the Y. pestis infection. These genes include unknown EST's, cytokines, enzyme of cytokine, receptors, ligands, transcriptional factors, inhibitor of transcriptional factor, and proteins involved with cytoskeleton. More interestingly, among them are 7 genes that encode for factors known to be associated with cell cycling and cell proliferation, with 3 of them playing a role in apoptosis. Our data also indicate that macrophage cells undergo apoptosis during an infection with Y. pestis-37 degrees C, however an infection with 26 degrees C cultures results in a delayed apoptosis. The correlation between the delayed apoptosis and the up-regulation of anti-apoptotic gene is currently being studied. PMID- 12756750 TI - Defensive function of phagocytes in pseudotuberculosis. PMID- 12756751 TI - Mechanisms of Yersinia enterocolitica evasion of the host innate immune response by V antigen. PMID- 12756752 TI - Yersinia pseudotuberculosis, its toxins and plant cells. PMID- 12756753 TI - Influence of Yersinia pseudotuberculosis on the immunity of echinoderms. PMID- 12756754 TI - Acute and chronic experimental infection processes caused by Yersinia pseudotuberculosis and evaluation of interleukin action on their development and outcome. PMID- 12756755 TI - Role of apoptosis of phagocytic cells in the development of immunodeficiency in plague. PMID- 12756757 TI - O antigen gene clusters of Yersinia pseudotuberculosis. PMID- 12756756 TI - Molecular genetics, biochemistry and biological role of Yersinia lipopolysaccharide. AB - Lipopolysaccharide (LPS) is the major component of the outer leaflet of the outer membrane of Gram-negative bacteria. The LPS molecule is composed of two biosynthetic entities: the lipid A--core and the O-polysaccharide (O-antigen). Most biological effects of LPS are due to the lipid A part, however, there is an increasing body of evidence also with Yersinia indicating that O-antigen plays an important role in effective colonization of host tissues, resistance to complement-mediated killing and in the resistance to cationic antimicrobial peptides that are key elements of the innate immune system. The biosynthesis of O antigen requires numerous enzymatic activities and includes the biosynthesis of individual NDP-activated precursor sugars in the cytoplasm, linkage and sugar specific transferases, O-unit flippase, O-antigen polymerase and O-chain length determinant. Based on this enzymatic mode of O-antigen biosynthesis LPS isolated from bacteria is a heterologous population of molecules; some do not carry any O antigen while others that do have variation in the O-antigen chain lengths. The genes required for the O-antigen biosynthesis are located in O-antigen gene clusters that in genus Yersinia is located between the hemH and gsk genes. Temperature regulates the O-antigen expression in Y. enterocolitica and Y. pseudotuberculosis; bacteria grown at room temperature (RT, 22-25 degrees C) produce in abundance O-antigen while only trace amounts are present in bacteria grown at 37 degrees C. Even though the amount of O-antigen is known to fluctuate under different growth conditions in many bacteria very little detailed information is available on the control of the O-antigen biosynthetic machinery. PMID- 12756758 TI - Cloning and characterization of the Yersinia enterocolitica serotype O:9 lipopolysaccharide O-antigen gene cluster. PMID- 12756759 TI - Characterization of the lipopolysaccharide outer core biosynthesis of Yersinia enterocolitica serotype O:3. PMID- 12756760 TI - ECA-antibodies in antisera against R mutants of Yersinia enterocolitica O:3. PMID- 12756761 TI - Lipopolysaccharides of Yersinia. An overview. PMID- 12756762 TI - The core structure of the lipopolysaccharide of Yersinia pestis strain KM218. Influence of growth temperature. PMID- 12756764 TI - Properties of the temperate Yersinia enterocolitica bacteriophage PY54. PMID- 12756763 TI - Yersiniophages. Special reference to phi YeO3-12. PMID- 12756765 TI - Transposon mutagenesis of the phage phi YeO3-12. PMID- 12756766 TI - Analysis of enterocoliticin, a phage tail-like bacteriocin. PMID- 12756767 TI - Function and regulation of the Salmonella-like pmrF antimicrobial peptide resistance operon in Yersinia pseudotuberculosis. PMID- 12756768 TI - Porin from Yersinia pseudotuberculosis: cloning and analysis of primary structure. PMID- 12756769 TI - Pore-forming proteins of genus Yersinia. PMID- 12756770 TI - Regulation of O-antigen biosynthesis in Yersinia enterocolitica. PMID- 12756771 TI - Regulation of the Yersinia pestis Yfe and Ybt iron transport systems. PMID- 12756772 TI - Function and regulation of the transcriptional activator RovA of Yersinia pseudotuberculosis. PMID- 12756773 TI - Temperature and growth phase regulate the transcription of the O-antigen gene cluster of Yersinia enterocolitica O:3. PMID- 12756774 TI - Molecular epidemiology of Yersinia enterocolitica 4/O:3. PMID- 12756775 TI - Occurrence of Y. enterocolitica in slaughter pigs and consequences for meat inspection, slaughtering and dressing procedures. PMID- 12756776 TI - Molecular epidemiology of the five recent outbreaks of Yersinia pseudotuberculosis in Finland. PMID- 12756777 TI - Yersinia pestis from natural foci. PMID- 12756778 TI - A virulence study of Yersinia enterocolitica O:3 isolated from sick humans and animals in Brazil using PCR and phenotypic tests. PMID- 12756779 TI - Molecular virulence characteristics and kinetics of infection of Yersinia pseudotuberculosis isolated from sick and healthy animals. PMID- 12756780 TI - Detection of Yersinia enterocolitica in slaughter pigs. PMID- 12756781 TI - Mechanism of formation of a population level of virulence of Yersinia pestis. PMID- 12756782 TI - Polymerase chain reaction assays for the presumptive identification of Yersinia pestis strains in Georgia. PMID- 12756783 TI - Genetic (sero) typing of Yersinia pseudotuberculosis. PMID- 12756784 TI - Yersinia spp. in the environment: epidemiology and virulence characteristics. PMID- 12756785 TI - Molecular characterization of Yersinia enterocolitica 1A strains isolated from Buenos Aires sewage water. PMID- 12756786 TI - Molecular typing of Yersinia strains by pulsed-field gel electrophoresis and RAPD PCR. PMID- 12756787 TI - Bacteriocin susceptibility of clinical Yersinia strains. PMID- 12756788 TI - Molecular epidemiology of Yersinia pseudotuberculosis. PMID- 12756789 TI - Growth of Yersinia enterocolitica in Inegol meatballs. PMID- 12756790 TI - Molecular genetic typing of Yersinia enterocolitica serovar O:8 isolated in Japan. PMID- 12756791 TI - Prevalence and characterisation of yadA-positive Yersinia enterocolitica in pig tonsils in 1995 and 1999. PMID- 12756792 TI - Yersinia pseudotuberculosis in pigs and pig houses in Finland. PMID- 12756793 TI - Evaluation of pulsed-field gel electrophoresis (PFGE) for Yersinia enterocolitica molecular epidemiology investigations. PMID- 12756794 TI - F1-negative natural Y. pestis strains. PMID- 12756795 TI - Occurrence of Yersinia pseudotuberculosis in iceberg lettuce and environment. PMID- 12756796 TI - A rapid method for the detection of enteropathogenic Yersinia in routine diagnostics of yersiniosis and pseudotuberculosis. PMID- 12756797 TI - Molecular epidemiological characterization of Yersinia pseudotuberculosis circulating in different geographic areas of the Russian Federation. PMID- 12756798 TI - Second and third generation plague vaccines. PMID- 12756799 TI - Yersinia outer protein E, YopE. A versatile type III effector molecule for cytosolic targeting of heterologous antigens by attenuated Salmonella. AB - Many Gram-negative pathogens evade the host's immune response by utilizing a specialized protein secretion machinery, known as type III secretion system (TTSS). Virulence factors such as the Yersinia outer protein E (YopE) are delivered directly into the cytosol of target cells in a TTSS-dependent fashion. This unique translocation mechanism can be used by attenuated Salmonella carrier vaccines for the delivery of heterologous antigens fused to YopE into the MHC class I-restricted antigen processing pathway. In orally immunized mice, this novel vaccination strategy results in the induction of pronounced peptide specific cytotoxic CD8 T cell responses. PMID- 12756800 TI - Immunological characterisation of sub-units of the Yersinia type III secretion apparatus. PMID- 12756801 TI - A recombinant prototrophic Yersinia pestis strain over-produces F1 antigen with enhanced serological activity. PMID- 12756802 TI - Vaccination with plasmid DNA expressing the Yersinia pestis capsular protein F1 protects mice against plague. PMID- 12756803 TI - Evaluation of protective immunity induced by Yersinia enterocolitica type-III secretion system mutants. PMID- 12756804 TI - Epidemiology and diagnostics of Yersinia-infections. PMID- 12756806 TI - Food-PCR. Validation and standardization of diagnostic PCR for detection of Yersinia enterocolitica and other foodborne pathogens. PMID- 12756805 TI - Susceptibility to plague of the rodents in Antananarivo, Madagascar. PMID- 12756807 TI - A multiplex PCR-detection assay for Yersinia enterocolitica serotype O:9 and Brucella spp. based on the perosamine synthetase gene. Application to Brucella diagnostics. PMID- 12756808 TI - Vascular endothelial growth factor in yersiniosis. A study on 157 military recruits. PMID- 12756809 TI - Pathogenic role of a superantigen in Yersinia pseudotuberculosis infection. PMID- 12756810 TI - Chronic infection with Yersinia enterocolitica in patients with clinical or latent hyperthyroidism. PMID- 12756811 TI - A new selective medium provides improved growth and recoverability of Yersinia pestis. PMID- 12756812 TI - Evaluation of a one-step biochemical screening test to determine pathogenic strains of Yersinia enterocolitica. PMID- 12756813 TI - Production of polyvalent Yersinia enterocolitica bacteriophage preparation for medical prophylactic use. PMID- 12756814 TI - New approaches to detect and assess the pathogenicity of clinical strains of Yersinia spp. based on molecular biology techniques. PMID- 12756815 TI - [Physicians' participation in clinical drug trials]. PMID- 12756816 TI - [Administrative organization and responsibility]. AB - The paper describes the current organisation of clinical trials in Danish hospitals, with particular emphasis on the relationship between hospitals and the pharmaceutical industry. Legal responsibilities as well as mutual agreements on collaboration and organisation are described and discussed. PMID- 12756817 TI - [Openness in the cooperation between physicians and private corporations]. AB - The past decades in Danish health services have been characterized by limited cooperation between health authorities, health employees, and private corporations. Today, society and legislature acknowledge the need to support this cooperation, which creates a great challenge to hospital management and its responsibility to create and secure strong research environments. This effort is often challenged by the tendency of the media to transform initial conflicts into problems of credibility. Society is also aware of the fact that increased cooperation is important to maintain the ability to develop new and better methods of treatment. It is therefore of vital importance to solve potential conflicts before the media interpret the problems, by encouraging openness in the cooperation between physicians and private corporations. PMID- 12756818 TI - [Publication ethics, national and international regulations]. AB - The publication ethics cover all stages of the scientific manuscript processing. In this respect a couple of ethical assumptions must be taken into account: the research ethics (i.e. respect and safety for participating patients and healthy volunteers), and the researcher's ethics (i.e. the credibility of the researcher). International declarations, conventions, directives, and various national laws and rules regulate research ethics and researcher's ethics. These topics are to some extent included in the Vancouver regulations, which also include qualifications for fulfilling authorship. PMID- 12756819 TI - [New drugs and European procedures of approval. The European Agency for the Evaluation of Medicinal Products' role]. AB - Whereas regulatory drug approval in Europe previously was the responsibility of national authorities, innovative new drugs are now being approved by the European Commission instead. Through its agency for drug evaluation (EMEA), the Commission provides guidance on the clinical documentation necessary to obtain regulatory approval for new drugs. As a large proportion of clinical drug trials is sponsored by industry and aims at obtaining regulatory approval, the guidance provided by EMEA will have increasing influence on clinical drug research in Europe. PMID- 12756820 TI - [Contact and dialogue between drug companies and the global regulatory authorities throughout research and development phase]. AB - Contact between pharmaceutical companies and local as well as global regulatory authorities (Ministry of Health) takes place throughout the development phase and especially when a clinical study is to be initiated. This contact is important both for the company and for the authorities, as both parties are constantly kept informed about the development progress and about the potential unexpected findings in connection with development of a new drug. The authority dialogue secures that authorities and companies exploit their resources optimally and patients get access to new, safe and efficacious drugs without delay. PMID- 12756821 TI - [The role of the Danish Medical Products Agency in clinical trials]. AB - The registration and legislation concerning clinical trials are described with specific focus on subjects of importance to Danish doctors. Especially the investigators' responsibility for the registration of the trial, reporting of serious adverse events and final report to the agency are mentioned. The fact that the evaluation of a protocol will focus on the risk, the contents of essential new knowledge and the design are emphasised. The process of approval and the fees are mentioned, and finally the changes caused by the new GCP directive are discussed. PMID- 12756822 TI - [The inspection by the Danish Medical Products Agency of drug clinical trials]. AB - An official review by the regulatory authorities of the conduct of a clinical trial is called a GCP-inspection and is an important element of Good Clinical Practice. The Danish Medicines Agency initiated GCP-inspections in Denmark in 1992. The article gives a short description of the inspections together with a few examples of findings during the inspections. PMID- 12756823 TI - [ICH-GCP Guideline: quality assurance of clinical trials. Status and perspectives]. AB - Good Clinical Practice (GCP) is an international ethical and scientific quality standard for designing, conduction, recording and reporting trials that involve the participating of human subjects. Since 1997 the ICH-GCP Guideline has been a requirement for conducting clinical trials which should be used as documentation to the authorities. The 13 fundamental principles of the ICH-GCP Guideline for conducting clinical trials are described. Furthermore, the most essential responsibilities of the investigator, sponsor and sponsor-investigator are mentioned. Investigator-initiated trials do not need to be conducted according to the GCP-principles but after the implementation of the EU Directive 2001/20/EC this will be changed. Today the GCP-principles only apply to clinical research with drugs but within a few years the GCP-principles will probably be a requirement for all clinical research. PMID- 12756825 TI - [How to establish GCP-units in Denmark?]. AB - According to a new EU Directive investigator initiated drug trials are to comply with the guidelines for Good Clinical Practice (GCP) as of May 2004. This implies that trials should be conducted according to a set of Standard Operating Procedures (SOPs) and be subject to monitoring and auditing. In 2001, investigators in Denmark initiated 73 drug trials. In order to provide GCP services and guidance to the investigators it is proposed to establish 3-4 regional GCP units at the three university hospitals. The estimated annual cost for the 3-4 GCP units will be between DDK 7.5 and 10 million. PMID- 12756824 TI - [The GCP directive--consequences for clinical drug research]. AB - The contents and implications of the EU Directive on good clinical (research) practice (GCP) regarding drug trials are described. As of May 2003, clinical researchers in Denmark must have standard operation procedures, conduct monitoring, consider quality assurance, and expect inspections. The industry may be better prepared, but the Directive makes GCP part of the law and phase IV studies become subject to GCP. Patients will be assured the same quality in trials irrespective of the industry or investigator being the sponsor and may look forward to quality improvement of drug trials. PMID- 12756827 TI - [Pharmacoeconomics--survey and status]. AB - Pharmacoeconomics is a subdiscipline of health economics. There are two main areas of application: economic evaluation (cost-effectiveness, cost-utility and cost-benefit analysis) and regulation of the drug market, e.g. price control, reimbursement mechanisms linked to prices (reference prices), parallel trade, research and development (R&D), and patents. It is shown how price regulation has led to price stability in the Danish drug market with a price level on or [table: see text] slightly below the European level. The basic principles of cost effectiveness analysis are outlined followed by a critical discussion of the use of guidelines in connection with application for reimbursement. It is noted that many methodological problems still have not been settled definitively leading to many low quality economic evaluations. PMID- 12756826 TI - [Public GCP unit--experiences from the Aarhus University Hospital]. AB - According to the EU GCP-directive, all clinical trials involving medical products must adhere to the GCP-standards by May 2004. This may entail difficulties for investigator-initiated trials not sponsored by the industry. A public GCP-unit was established at Aarhus University Hospital in 1995. The unit offers researchers a monitoring system including discussion of the protocol, check of formalities, and verification of data and protocol compliance. As a contribution to the future work with GCP, experience from the GCP-unit at Aarhus University Hospital is presented here. PMID- 12756828 TI - [Does the public sector have an independent research role in the development of drugs?]. AB - Exclusively private companies do drug development. The State contributes with education of academics and basic research constituting the basis of half of the drugs developed by the private companies. The Danish private drug research amounts to six billion DKK per year, corresponding to the estimated price of the development of one new drug. The development shows a negative tendency. There are doubts about the scientific credibility, the number of new drugs is declining, drug development costs are rising, and the competitiveness in Europe is declining compared with the one of The United States. Continued improvement of Danish drug development can be achieved by stimulation of the public research related to drug development. PMID- 12756829 TI - [The roles and responsibilities of the pharmaceutical industry]. AB - A major part of clinical research in Denmark involves clinical testing of pharmaceuticals sponsored by the pharmaceutical industry. All these trials are carried out according to Good Clinical Practice (GCP) and necessitate a close working relationship between responsible investigators and the pharmaceutical industry. It is of mutual interest that these trials should have a high scientific standard and that the integrity of patients always has the highest priority. Many guidelines, laws and conventions regulate this area to ensure the fulfilment of these goals. In the present article, we describe the roles and responsibilities that pharmaceutical companies have to comply with. PMID- 12756830 TI - [The scientific committee system]. AB - Since 1992 a law based national system of Scientific Ethical Committees has existed. It consists of regional committees supplemented with a Central Committee. The Danish model is characterised by having a majority of lay people in all committees. The law is based on the principle of informed consent. A new law in 2003 will implement the EU directive on Good Clinical Practice. A maximum time of 60 days for the approval of biomedical research projects and approval of multi-centre trials at only one scientific ethical committee will be introduced. PMID- 12756831 TI - [The personal data act, clinical trials and data privacy. Rules for treatment of personal data in clinical trials and scientific research projects]. AB - The Danish Act on Processing of Personal Data applies to all processing of data for the purpose of carrying out medical trials and other scientific or statistical studies. Prior to the commencement of the processing of data, a private controller, i.e. a pharmaceutical company, shall notify the Danish Data Protection Agency and obtain the authorisation of the Agency. The article presents the various conditions laid down by the Agency for the carrying out of the processing operations. Furthermore, special attention is drawn to the data subject's various rights. In order to ensure the privacy of the data subject and to protect data against accidental or unlawful destruction, loss or alteration and against unauthorized disclosure, the controller shall implement certain technical and organisational security measures laid down by the Agency. When a controller leaves the processing of data to a processor, the processor is obliged to ensure compliance with the given measures. The Data Protection Agency supervises that the processing is carried out in compliance with the provisions of the Act and the conditions laid down by the Agency. PMID- 12756832 TI - [What are the requirements for establishing a biological specimen bank?]. AB - Establishing of a biological bank requires consideration. Optimal location of the bank for handling of the samples and quick response to alarms are essential. Decisions about freezer system (electrical or liquid nitrogen container), how to store the biological material (tubes or straws) as well as proper labelling of the samples are important. Quality control must be considered to monitor possible degradation over time. PMID- 12756833 TI - [Attitude of patient associations to clinical trials of new drugs]. AB - Both the Danish Rheumatism Association and the Danish Cystic Fibrosis Association think that patients have a right of access to new and better medicine. The patients affiliated to these two patient groups are more than willing to enroll in clinical trials on new drugs. The Danish Rheumatism Association focuses on good information and security while the Danish Cystic Fibrosis Association has an unrestricted willingness to enroll in clinical trials. PMID- 12756834 TI - [Dishonesty in clinical trials is expensive for the society]. PMID- 12756835 TI - [How long does a medical truth survive?]. PMID- 12756836 TI - [Obstetric acupuncture]. PMID- 12756837 TI - [Condom against HIV infections: no point]. PMID- 12756838 TI - [Renal artery embolism]. AB - Renal artery embolism (RAE) is a rare disease. Urgent treatment is necessary, as ischaemia can cause irreversible kidney damage in 60 to 90 minutes. RAE frequently clinically manifests as a pain similar to renal colic. Source of embolus is predominantly the heart at atrial fibrillation. Laboratory findings are unspecific. Ultrasonography with color Doppler imaging is essential. Kidney perfusion is low and upper urinary tract is undilated. Renal function can be recognized by intravenous urography and at renal scintigraphy. In angiography, renal artery is closed with thromboembolus. With no delay, transcatheter clot aspiration should be performed and fibrinolytic agents (tissue plasminogen activator) should be topically administered. Continual heparinisation and later warfarinisation should follow. In spite of successful revascularisation, parameters of kidney function can almost never reach that prior the RAE and shrinkage of kidney becomes a frequent consequence. Treatment can be successful even in patients with renal occlusion lasting over 90 minutes, since occlusion is often incomplete or significant collateral blood supply exists. In conclusion, renal artery embolism must be considered in cases of flank pain in patients with certain risk actors (especially atrial fibrillation). Ultrasonography with color Doppler imaging and urgent angiography of the renal artery are necessary in these cases. Thromboembolus can be then aspirated, and kidney perfused with fibrinolytic agent. PMID- 12756839 TI - [Somatostatin, somatostatin analogs and their clinical use]. AB - Somatostatin is a hormone with broad spectrum of physiological effects. Somatostatin and its analogues are employed in the diagnosis and treatment of various clinical conditions. The article reviews the use of somatostatine and somatostatine analogues in the clinical medicine. It also comprises the overview of the basic therapeutical and diagnostic indications for somatostatine and its analogues and the theoretical basis of their use. PMID- 12756840 TI - [Markers of lung disease in expired air]. AB - Many chronic inflammatory pulmonary diseases are associated with inflammation and oxidative stress. A complex interplay between the specific cause of the diseases, the type and intensity of inflammation and oxidative stress, results in the clinical picture and reveals the probability of progression of the specific disease. The presence of inflammation and oxidative stress has been established in interstitial lung diseases, cystic fibrosis, bronchiectasis, adult respiratory distress syndrome, chronic obstructive lung diseases and asthma. Noninvasively obtained markers of inflammation and oxidative stress from the breath may assist in diagnosis of pulmonary diseases, assessment of diseases severity and response to treatment. Exhaled markers include NO, CO, H2O2, ethane, pentane, and isoprostanes. PMID- 12756841 TI - [Multiple sclerosis and magnetic resonance]. AB - Multiple sclerosis is a demyelinating process presently referred to autoimmune diseases. Its diagnostics is based on clinical examination and paraclinical tests (magnetic resonance, examination of CSF and evoked potentials recording). Magnetic resonance (MR) has the highest significance, both for the diagnostics and for the monitoring of the course of disease and results of treatment. Results of magnetic resonance are not specific for the multiple sclerosis and therefore for the reliable diagnosis the McDonadl's criteria have to be fulfilled. It appears that magnetic resonance is more sensitive to progression of disease than the clinical examination. Monitoring of the course of disease requires new techniques of MR imaging. Automatic, software assisted determination of plaque volumes in T2 and T1 weighted images--so called "lesion load", is checked during the patient's treatment. Assessment of brain volume determines progression of atrophy. The aim of all the new methods of MR imaging is to search for a reliable technique of the disease monitoring and namely for the prediction of disease progression. PMID- 12756842 TI - [Disease-causing mutations versus neutral polymorphism: use of bioinformatics and DNA diagnosis]. AB - Molecular genetic diagnostics is available for increasing number of genetically determined diseases. A wide spectrum of mutations can be detected by laboratory methods. A mutation can be defined as a change in a specific DNA sequence when compared with the reference sequence published in the gene database. However, in some cases it is difficult to distinguish if the detected sequence variant is a causal mutation or a neutral (polymorphic) variation without any effect on phenotype. The interpretation of rare sequence variants of unknown significance detected in disease-causing genes becomes an increasingly important problem. Further analysis on DNA and on protein levels with the use of bioinformatics are needed to reveal the effect of rare sequence variants. Inherited complex disorders, for example rare hereditary forms of cancer diseases, represent a challenge to molecular geneticists. The identification of exact causal mutation directly responsible for the development of the disease and for the assessment of disease risk resulting from this genetic variation has further implications. Predictive genetic diagnostics allows identify relatives at high risk of genetically determined disease and use of targeted preventive and therapeutic approaches. In severe cases it allows also prenatal or pre-implantation diagnostics. PMID- 12756843 TI - [Intravascular brachytherapy in the prevention of vascular restenosis]. AB - Vascular restenoses currently represent a major problem in the treatment of vascular stenoses. One of new approaches in the prevention of restenoses is intravascular brachytherapy. Intravascular brachytherapy uses local irradiation of the stenotic vessel segment by ionizing radiation with the aim of prevention of restenosis. This is a new rapidly developing multidisciplinary approach based on collaboration of specialties of intervention radiology, intervention cardiology, angiology, nephrology and radiation oncology. This review examines current options of intravascular brachytherapy as well as results of clinical trials evaluating the efficacy of intravascular brachytherapy in the different anatomical regions. Intravascular brachytherapy may substantially reduce the rate of restenoses. However, intravascular brachytherapy should be currently used only in the setting of clinical trials. Optimal method of irradiation of the stenotic segment of the vessel is still to be defined. PMID- 12756844 TI - [Hormones and quality of life in aging men]. AB - BACKGROUND: Aging men suffer from a decrease of androgen-anabolic steroids known as "Partial Androgen Decline in the Aging Male" (PADAM) that is sometimes compensated by the use of androgen replacement therapy. METHODS AND RESULTS: To decide whether androgen therapy should be considered, hormonal serum levels are commonly tested. In addition, the St. Louis questionnaire is used as and indication. Accordingly, serum levels of testosterone, androstendione, dihydrotestosterone, dehydroepiandrosterone, its sulphate, epimers of 7-hydroxy dehydroepiandrosterone, epitestosterone, lutropin, follitropin, prolactin and sex hormone binding globulin were measured in 216 men over 50 years of age. Further, the Sr. Louis questionnaire was applied and the extent of the relations among the data was evaluated. RIA and IRMA kits from Immunotech and Orion-Diagnostica were used for measurement of the hormones except epitestosterone and 7-hydroxy dehydroepiandrosterone epimers, which were determined using specific radioimmunoassays. CONCLUSIONS: The decline in the index of free testosterone, given by the increase of SHBG levels, as well as the decrease in the levels of DHEA and its derivates were the most prominent. No correlation was found between the levels of hormones and the individual items of the questionnaire or with the total score calculated from the questionnaire data. In conclusion, the University of Saint Louis questionnaire did not give a reliable indication of an androgen deficit. However, the found values and their changes can still be helpful during the decision process concerning indication and initiation of hormone replacement. PMID- 12756845 TI - [Would ultrasonography contribute to the early detection of clinically silent dialysis access stenoses?]. AB - BACKGROUND: Stenoses are leading factors limiting dialysis vascular access patency. Both physical and haemodynamic examinations are used in stenoses screening. To estimate potential advantage of ultrasonography as a screening measure, we calculated both sensitivity and specificity of clinical (nephrological) diagnoses of access stenoses compared with ultrasound. METHODS AND RESULTS: 268 examinations of 193 subjects were included into this comparison. Linear-array 7.5 MHz ultrasound probe were used for whole-length access examination. Sensitivity of clinical examination was 35.8%, specificity 92.8%. Sensitivity did not differ significantly according to the stenosis localization. CONCLUSIONS: Ultrasound use in dialysis access screening would considerably increase the number of stenoses diagnosed in-time. On the contrary, clinical suspicion on access stenosis is highly specific, therefore it is not indispensable to confirm it by ultrasound. PMID- 12756846 TI - [Dipeptidyl peptidase IV (DPP IV, CD 26): a tumor marker in cytologic and histopathologic diagnosis of lesions of the thyroid gland]. AB - BACKGROUND: Morphological diagnostics of thyroid gland tumours faces certain differential diagnostic problems. Extensive histological examination of the entire tumour is required for the final diagnosis of follicular and oncocytic tumours. Thus, assessment of reliable definitive cytological and/or intraoperative histological diagnosis is not possible. No marker of malignancy has been so far generally accepted in the thyroid tumour diagnosis. The aim of the study was to evaluate membrane protease dipeptidyl peptidase IV (DPP IV) in the differential diagnosis of thyroid tumours. METHODS AND RESULTS: DPP IV was assessed cytochemically in 254 smears, histochemically in 314 cryostat sections, and immunohistochemically in 309 paraffin-embedded sections obtained from the group of 336 patients. There were 283 females and 53 males with the mean age of 48 years (range 15-80 years) in this series. Sensitivity of cytochemical detection was 71%, specificity was 96%, and diagnostic accuracy was 93% using the 50% threshold. Histochemically, sensitivity was 71%, specificity was 93%, and diagnostic accuracy was 90% using the 5% threshold. Using the immunohistochemical assessment, sensitivity was 68%, specificity was 94%, and diagnostic accuracy was 91% using the 5% threshold. CONCLUSIONS: According to our results, DPP IV can be used as a marker of malignancy in well-differentiated carcinomas of follicular cell origin, namely in papillary carcinoma. However, it is less reliable in follicular and oncocytic carcinomas. PMID- 12756847 TI - [Fluorescence cystoscopy using 5-aminolevulinic acid]. AB - BACKGROUND: Fluorescence diagnosis has an increasing importance in different medical fields. In the presented paper, comparison of cystoscopy in white light with fluorescence cystoscopy after intravesical administration of 1 g of 5 aminolevulinic acid is presented. METHODS AND RESULTS: From the group of 63 persons examined, no difference between findings in white and blue light was found in 39 cases. In 21 patients more pathological spots were found in blue light (10 cases) or, in agreement with histology, pathology was detected in the blue light only (11 cases). CONCLUSIONS: The intravesical administration of 5 aminolevulinic acid had no side effects. Our study has definitely proved the advantages of fluorescence cystoscopy. PMID- 12756848 TI - [CD14 (C-159-->T) polymorphism and levels of C-reactive protein]. AB - BACKGROUND: High plasma levels of C-reactive protein (involved in unspecific host defence) have been described as independent risk factor of atherosclerosis. The possible association has been analysed between the CD14C-159-->T polymorphism and plasma level of CRP. METHODS AND RESULTS: With the PCR and subsequent restriction analysis we have evaluated C-159-->T polymorphism in the CD14 gene of 166 representatively selected male Caucasians with known plasma level of CRP. The CRP was measured immunologically. A significantly higher (p < 0.01) frequency of the CD14-159TT homozygotes between the individuals with the plasma level of CRP > 2.19 mg/l (31.9%, 15 out of 47) has been detected when compared to the group with plasma level of CRP lower than 2.18 mg/l (11.9%, 5 out of 42). CONCLUSIONS: CD14 C-159-->T polymorphism could be the first described genetic marker associated with plasma level of C-reactive protein. PMID- 12756849 TI - [New drugs for treatment of parasitic infections]. AB - In this review we have summarized published data on two new compounds, which can represent important antiparasitic drugs in the near future, nitazoxanide for treatment of intestinal parasitic infections including cryptosporidiosis and miltefosine for oral treatment of visceral leishmaniasis. PMID- 12756850 TI - [Endobronchial neurofibroma]. AB - Neurofibroma belongs to the benign tumors and it is categorised as neuroectodermal tumor. Neurofibromas are most frequently found in the posterior mediastinum, their endobronchial localisation is rather rare. Our paper presents two cases of endobronchial neurofibromas with diverse clinical and X-ray symptomatology. Both patients underwent surgery and the etiology of tumours was assessed by histological examination of the preoperative biopsy. PMID- 12756852 TI - Genes for human behavior. PMID- 12756853 TI - The challenges of genetic tests for human behavior. AB - OBJECTIVE: To review the potential benefits and adverse effects of genetic tests that may develop, that effectively predict mental illness or variation from the norm on behavior traits. METHOD: A review is offered of the history of genetic based political policies and racial discrimination. The current status of genetic research in mental disorders is described. Anticipated genetic progress, and derivative tests and limits on their predictive power are considered. RESULTS: There does exist a potential for invidious stigmatization and discrimination, based on genotypes of individuals and allele frequencies in communities, in various transactions such as health care access, employment and education, as well as in more intimate decisions including choice of spouse and decisions to become a parent. CONCLUSIONS: The major uses of valid genetic tests for human behavioral traits will be for development of knowledge on pathophysiology of illness, and for development of new treatment and educational strategies to deal with illness. Some extension of genetic tests into predictions about specific individuals may well occur. Awareness of the potential for discrimination, and a determination to develop culturally sensitive and fair applications of genetic tests, is required to avoid adverse effects on individuals and communities. PMID- 12756855 TI - Genetics of complex psychiatric disorders: scientific foundations. AB - Advances in genetic analysis as well as progress in the Human Genome Sequencing Project have raised the hope to elucidate the molecular basis of mental disorders on the DNA level. In the present review we describe and discuss the basic concepts which are currently applied for the genetic approach in order to find DNA variations associated with a disorder. A prerequisite for such studies is the confirmed evidence for genetic transmission established by family-, twin-, and adoption studies. We describe the study design and discuss the mode of inheritance for complex genetic traits in comparison to monogenic, Mendelian traits. Recombination as a basis for linkage analysis is explained and its implication for classical LOD score analysis in families, as well as for affected sib-pair analysis and for analysis of linkage disequilibrium is discussed. Moreover, we describe types of markers and maps used in these studies. We explain and discuss the association approach in conjunction with the Human Genome Project. Finally, we stress possible implications for diagnosis, prevention and new therapeutic approaches. PMID- 12756854 TI - History of the eugenic movement. AB - The goal of this review is to introduce the reader to the ideas behind the eugenic movement, its implementation, and its consequences. First, we address the work of prominent 19th century forerunners of eugenics. Second, we discuss the eugenic movement during the first half of the 20th century, and its common and differential characteristics in countries where its expression was more marked, including the US and Nazi Germany. We also discuss the eugenic movement in Sweden, whose history has remained unknown until recently. We analyze the reasoning behind forced sterilization and involuntary euthanasia. Finally, we consider the impact that past eugenics has had on the profession of psychiatry and psychiatric research, particularly genetics. We argue that the history of the 20th century eugenics movement and its leaders still require attention. PMID- 12756856 TI - Genetics of schizophrenia: a review of linkage findings. AB - BACKGROUND: Family, twin and adoption studies have demonstrated the genetic basis of schizophrenia. Several genes, acting synergistically with each other and with the environment probably underlie the disorder. Different genes for schizophrenia are surely present in different populations. Parametric and nonparametric linkage analyses are the main methods employed in the search for schizophrenia genes. Only one study, published recently, has aspired to report the cloning of such a gene, and its significance is debated. METHOD: This review covers the significant and suggestive evidence found in linkage studies for localization of schizophrenia genes. Support for these results from other sources is also described. RESULTS: Significant and suggestive linkage results were reported for at least 18 different chromosomal regions. The more convincing evidence is for loci on 1q, 5q, 6p, 6q, 8p, 10p, 13q, 22q and Xp. CONCLUSIONS: Current methods of genetic analysis are limited in their power to detect genes for complex disorders such as schizophrenia. Technological advances and the use of special populations such as genetic isolates could overcome these limitations. PMID- 12756857 TI - Genetics of suicidal behavior: candidate association genetic approach. AB - Suicidal behavior runs in families and seemed to be genetically determined in part and independent of the presence of psychiatric disorders. This review presents the current knowledge of candidate gene association studies in the field of suicidology. Concordance for monozygotic twins is about 13% Vs. 0.7% in dizygotics. It seems that there is a relationship to intermediate phenotypes such as impulsivity and aggression. The problem of studying complex traits and of ethnic stratification in heterogeneous population are major concerns in studying genetics of suicide. Family-based methods is one of the strategies to overcome stratification. We found a possible association of tryptophane hydroxylase gene polymorphism with depression and serotonin transporter promoter gene to violence traits by family-based methods. Recent data from the study of COMT. MAO and serotonin receptors genes polymorphisms are controversial at this stage. Future directions for research includes alternative phenotypes, endo-phenotypes and genome scan. PMID- 12756858 TI - Heritability, genetics and association findings in anorexia nervosa. AB - Anorexia nervosa (AN) is a severe psychiatric disorder, characterized by a combination of abnormal eating behavior and weight regulation with disturbances of attitudes toward body weight and shape. Prevalence is estimated at 1/1000, but with a high prevalence of the partial syndrome and a 10% mortality rate. This article reviews the findings concerning the heritability and the contributing genes of the disorder, with a focus on candidate genes. In family studies, a higher frequency of AN and BN was found among relatives of AN probands. The heritability rate was estimated at 0.71, similar to twin studies, which estimate 0.58-0.76. The search for genes responsible for the disorder focuses on the monoaminergic and peptidergic systems that are related to appetite and weight regulation. So far, for serotonin and dopamine there are no consistent findings in association studies of AN. However, in an Israeli study, an association was found between susceptibility for AN and the COMT gene, which is involved in monoamine metabolism. Another Israeli study found a relation between AN and the gene encoding for the potassium channel (hSKCa3), which is involved in regulation of neuronal activity. In the endocrine system an unequivocal finding was described of an association to the gene encoding the receptor for beta-estrogen. In the appetite and weight regulation system an association was described between AN and a marker of the uncoupling protein-2 and -3 chromosomal region, raising the likelihood that the mutation within the gene is close to a positive marker. To conclude, although there is a strong familial component in AN, so far the search for candidate genes has not been fruitful and further large scale prospective and adoption studies are needed to confirm genetic factors. We hope that relative studies using a wide genome scan, as well as subtyping the different types of AN, will bring us closer to understanding of the heritability of AN and enable the development of improved means of prevention and treatment. PMID- 12756859 TI - Personality genetics, 2002. AB - The study of the genetics of personality looks for effects of genetic differences on psychological differences between people. The most replicated associations have been between the dopamine D4 receptor gene and "novelty seeking" and between the serotonin reuptake transporter gene and anxiety. But even these reports are controversial, and their effects are too small to add to existing scientific theories or allow for clinical use in psychological counseling. Simultaneously studying three polymorphisms suggests that gene-gene interactions are partly responsible for these difficulties. Recently a linkage of anxiety-related traits to chromosome 8p21-23 has been replicated, and we may be close to isolating relevant genes in the region. But uncertainty about psychological phenotypes remains, and it is not clear whether future breakthroughs will require studies of thousands of genes in very large samples, or revolutions in the current assumptions and methods of personality genetics. PMID- 12756860 TI - [Patient and family oriented information and counseling in "home nursing of sick children" in Germany--a stepchild in scientific nursing research?]. PMID- 12756861 TI - [The role of nursing in public health/health promotion--attempt at a systematic approach]. AB - Public health and nursing meet mainly in the fields of care managing processes and health promotion. Since there is a new development of Public Health in the German speaking countries also the role of nurses in health education and health promotion has become more important. But traditional fields and new conceptions of public health nursing are still unmethodical. I present a systematic approach of different fields and different strategies of health promotion in nursing which has to be discussed further on. Health promotion in nursing is differentiated in a behavioural approach (empowerment of patients on different levels of nursing) and an approach which focuses on the conditions for health, mainly those for better social support, for better health in the community and in other settings. PMID- 12756862 TI - [Prevention of decubitus ulcer: theory and practice]. AB - In Spring 2001, the Department of Nursing Science, Humboldt-University in Berlin, conducted a pressure ulcer prevalence study on 3012 patients of eleven hospitals. The investigation revealed the use of pressure ulcer prevention on patients with different stages, or without a pressure ulcer. The study also focused on the use of interventions and bed-aids on patients both at risk and not at risk. Interventions without the use of bed-aids were used more often than interventions using bed-aids. The main type of intervention used was the mobilisation of the patient. This was found in all groups (at risk or not at risk; with or without a pressure ulcer). The bed-aids which were most frequently used were pressure relieving cushions and heel-protectors. Because of the differences in pressure ulcer management among the hospitals, this article also shows evidence on bed aids and the interventions as known from literature. In four of the interventions as used in practice, the highest degree of evidence could be found. However, in five cases no evidence could be found at all. The comparison of the use in practice and the literature revealed that interventions which are often used as pressure reduction are well tested and can be recommended. However, the (sheep-) skins, which are frequently used, are not reducing the pressure significantly. PMID- 12756863 TI - [What decision-making authority regarding oral care do nurses have?]. AB - By means of a written questionnaire nurses of different wards of various disciplines in 21 German hospitals were asked about their decision-making authorization in mouth care. At the same time experience in this field of nursing was examined. The study revealed that nurses have a high autonomy concerning their decisions in performing mouth care. They decide independently on necessity, indication, and aids to be applied. They are also very independent when it comes to the evaluation of the interventions' effect. Finally, they are the professionals to inform and educate patients on the subject of mouth care. Furthermore, most nurses are not satisfied with the co-operation between nursing and dental medicine. Even in the case of serious mouth problems a dentist's assistance is only rarely sought for although nurses consider such help necessary. Increased co-operation between nurses and dentists is therefore recommended to improve mouth care in hospitals. PMID- 12756864 TI - [Some frequently used substances, instruments and methods in mouth care with regard to evidence-based applications]. AB - The purpose of this study was to evaluate current practice in mouth care as few data exist in this area. By means of a questionnaire nurses were asked which mouth care substances, methods and instruments they used in their clinical practice. They were also asked about their clinical experience. The most frequently used substances and instruments were then examined with regard to their evidence base using an evaluation system bases on a compilation of scientific literature. The results show that nurses' selections of mouth care substances and instruments are mainly based on their clinical experience and on traditions. There is little scientific evidence for their mouth care practice. Using assessment instruments in order to assess oral conditions and evaluate nursing interventions as well as using more substances with proven effect has the best potential to improve practice. There is an obvious need for evidence-based guidelines and standards. PMID- 12756865 TI - [Improving job morale of nurses despite insurance cost control. 1: Organization assessment]. AB - Faced with declining resources for health care and greater pressures to improve productivity of nursing staff, nursing administrators must act now to develop organizational responses to morale problems among nursing staff. As part of a two part series for JONA, the authors describe low-cost organizational approaches that address nursing morale. Presented in Part 1 is a low-cost diagnostic process for assessing needs of staff and appraising organizational dimensions contributing to morale. Assessment findings provide clear direction for developing organizational approaches for improving morale. PMID- 12756866 TI - Primary squamous oesophageal cancer. PMID- 12756867 TI - Penman Lecture. Fluid resuscitation--where, when, how much and what fluid. PMID- 12756868 TI - Preliminary report on the effect of brachytherapy on expression of p53, bc1-2 and apoptosis in squamous cell carcinoma of the oesophagus. AB - Pre-brachytherapy biopsies and post-brachytherapy oesophagectomy specimens of 10 patients with early squamous cell carcinoma of the middle third of the oesophagus were examined for the expression of p53, bcl-2 and apoptosis using immunohistochemical markers. There was no expression of p53 in one patient in both pre- and post-brachytherapy specimens. In 8 patients, p53 staining was strongly positive (3+) with approximately 50% or more cells, and with diffuse and no specific pattern in the pre-brachytherapy biopsies. The tumour areas of the post-brachytherapy specimens of this group showed strong 3+ positivity with p53 (10-50% positive cell count), with the pattern being focal and peripheral in the tumour islands. The centre of the tumour islands showed necrosis and/or keratinisation. In one patient, the pre-brachytherapy biopsy showed expression of p53 while the post-brachytherapy specimen was negative. bcl-2 expression in both pre- and post-brachytherapy was equivocal and inconclusive in both the pre- and post-brachytherapy specimens. Apoptosis was negative in all the pre- and post brachytherapy tissue sections in the presence of positive controls. Brachytherapy does not cause cell death by apoptosis but by necrosis and maturation of the cells into better differentiated cells, which is caused by OH free radical, and induction of the keratin gene respectively. It is possible that brachytherapy may cause destruction of cells containing wild-type p53, while mutant p53 in cells located at the tumour periphery escape the effect of brachytherapy. This may be responsible for the high incidence of local recurrence and distant metastasis in oesophageal cancer treated with radiotherapy. There is no effect of brachytherapy on bcl-2 expression in oesophageal cancer. PMID- 12756869 TI - Gastrointestinal stromal tumours of the oesophagus. AB - A rare case of gastrointestinal stromal tumour (GIST) of the oesophagus is presented. Pathological features with a review of the treatment options and the literature are presented. PMID- 12756870 TI - Why we use the donor left kidney in live related transplantation. AB - In addition to the superior graft survival afforded by live related transplantation, this option has assumed an important role in the management of endstage renal failure in centres confronted with a scarcity of cadaveric kidneys. In pursuing this option, it is imperative that the donor has minimal morbidity. An ongoing dilemma is which side the kidney should be harvested from. This study reviews the anatomical basis for selecting the left kidney and the impact on outcome for patient and donor. A database comprising cadaveric and clinical subsets was analysed. The total sample size analysed was 1 244 kidney pairs (305 cadaveric; 939 clinical (61 live related left kidney transplants harvested by the extraperitoneal approach)). RESULTS: Additional renal arteries (ARAs): Right first, second = 18.6%, 4.7%; left first, second = 27.6%, 4.4%. Additional renal veins (ARV): Right first, second = 26%, 3.3%; left first only = 2.6%. Length of renal vein (cm): Right 2.4 +/- 0.7, left 5.9 +/- 1.5. Other venous variations encountered were on the left side only (renal collar 0.3%, retro-aortic vein 0.5%). In the live related transplant series 24.6% ARAs were encountered (first 19.7%, second 4.9%). The postoperative course and outcome of both donor and recipient were not associated with increased morbidity. While greater length of the left renal vein (LRV) afforded easier technical manipulation, it is interesting to note that its length is shorter than that reported in the literature. ARVs are infrequent on the left and when encountered the smaller calibre vessel may be ligated with impunity due to rich intrarenal anastomosis. In selecting the donor kidney the surgeon has to balance the prospect of fewer ARAs on the right against the benefit of a longer LRV. The solution to this dilemma can only arise from a randomised clinical study. In our practice, consistent use of the left kidney has not affected clinical outcome. PMID- 12756871 TI - Gallstone ileus--clinical image. PMID- 12756872 TI - [A patient with an unexplained loss of consciousness: a case for the neurologist or the cardiologist?]. AB - Three patients, a woman aged 62 and two men aged 22 and 77 years respectively, were admitted because of an unexplained loss of consciousness. In the woman and younger man these complaints were caused by a vasovagal reaction, and in the older man by hypersensitivity of the carotid sinus. In a hospital setting, the examination of patients with a loss of consciousness often leads to discussions between the neurologist and the cardiologist. Elaborate additional testing seldom reveals epilepsy or cardiac arrhythmia as the cause. A thorough medical history can exclude both diagnoses in almost all cases. Physicians should be aware that most patients experience a vasovagal syncope and that this might be triggered by very common circumstances. Knowledge of these triggers and the epidemiology of loss of consciousness might prevent the patients from being examined by different specialists and undergoing unnecessary additional tests. PMID- 12756874 TI - [ 'Severe acute respiratory syndrome' (SARS) in perspective]. AB - In the Netherlands, the risk of an outbreak of the severe acute respiratory syndrome (SARS) appears to be limited, if people in contact with a possibly imported case observe strict measures to prevent contamination. SARS may well be a zoonosis. Bovine spongiform encephalopathy and outbreaks of virus infections in the bioindustry, such as swine fever, foot-and-mouth disease and classical avian influenza, have led to massive killing of cattle, swine and fowl in the Netherlands. The possibility that the bioindustry constitutes a risky microbiological 'experiment' makes a discussion as to its future in the Netherlands urgent. PMID- 12756873 TI - [Violent behavior in men due to genetic predisposition and childhood abuse: an hypothesis]. AB - Caspi et al. recently reported that the likelihood of an abused child becoming violent as an adult hinges on a gene regulating the expression of monoamine oxidase A. This enzyme metabolises neurotransmitters. The scientists followed 442 boys from birth to their mid twenties and found an interaction between childhood abuse and a genetic polymorphism. Major strengths of the study are the longitudinal design and the excellent follow-up rate. Moreover, childhood abuse and antisocial behaviour are operationalised in various manners. The different forms of childhood abuse and neglect are, however, summarised and individual results are only presented for the different outcomes. In this way the consistency of the results is shown, but significance is only reached for one outcome: 'antisocial personality disorder symptoms'. It is especially important to replicate the study that investigated an association but no causal mechanism. Thus, conclusions about susceptibility to childhood abuse or medical treatment options for neglected children are premature. PMID- 12756875 TI - [Initial diagnostic strategy in the case of transient losses of consciousness: the importance of the medical history]. AB - It is estimated that almost half of the people experience a transient loss of consciousness at some time during their life. In young patients (< 35 years) the cause is mostly a reflex syncope. In older patients the common causes are orthostatic and postprandial hypotension, sinus caroticus syndrome, cardiac arrhythmias and valvular disorders. The medical history can identify a probable cause of the transient loss of consciousness in almost all young patients (< 35 years) and in the majority of older patients. A physical examination and an ECG should be performed in all patients who have experienced a transient loss of consciousness, other than those with classical vasovagal syncope, in order to exclude orthostatic hypotension and dangerous cardiac causes. PMID- 12756877 TI - [Diagnostic image (137) An infant with fever and pancytopenia. Hemophagocytic lymphohistiocytosis]. AB - A 2-month-old male infant with consanguineous parents had fever of unknown origin, pancytopenia, and hepatosplenomegaly. Laboratory tests, peripheral blood smear and bone marrow revealed haemophagocytic lymphohistiocytosis. PMID- 12756876 TI - [Diaphragm dysfunction in patients with chronic obstructive pulmonary disease]. AB - Chronic obstructive pulmonary disease (COPD) is characterised by alterations in the airways and lung parenchyma resulting in an increased respiratory workload. Besides an increased load and hyperinflation of the thorax, additional factors, such as systemic inflammation, oxidative stress, hypoxia and loss of muscle mass, further have a negative influence on diaphragm contractility. The diaphragm seems to adapt only partly to the altered circumstances to which it is exposed. As a consequence, several morphological, biochemical and functional alterations occur in the diaphragm, resulting in diaphragm dysfunction. In an appropriately selected group of patients, the function of the diaphragm can be improved by respiratory muscle training, oral nutritional therapy or hormonal interventions. PMID- 12756878 TI - [Stereotactic large core needle biopsy for the diagnosis of nonpalpable breast lesions: reliable without additional excision biopsy]. AB - OBJECTIVE: To determine the reliability of stereotactic large core needle biopsy (SLCNB) in the diagnostic work-up of suspected carcinomatous non-palpable breast lesions. DESIGN: Prospective. PATIENTS AND METHODS: Between 1 February 2000 and 6 June 2002, data were collected on all patients with a non-palpable breast lesion who were scheduled to undergo a SLCNB. SLCNB procedures were performed in 4 centres and patients were referred from 40 hospitals in the Netherlands. Histological diagnosis at SLCNB and subsequent therapy were recorded. Results were compared with results of the preceding 'Core biopsy after radiological localisation' (COBRA) study, where each large core needle biopsy was followed by an excision biopsy. Follow-up data were also obtained from the Dutch National Automated Pathology Archive (PALGA). The adherence to the COBRA guidelines was also evaluated. RESULTS: Nine hundred and fifty-five patients were included with 995 lesions, and 905 biopsy procedures were completed in 874 patients (mean age: 59 years; range: 23-86). Of the high-risk lesions, 27% were found to be carcinomas upon excision biopsy, which is comparable to the results of the COBRA study (23%). Twenty-eight percent of the ductal in-situ carcinomas were found to be invasive, which is higher than in the COBRA study (17%). No malignancies were found during the limited follow-up period (mean: 20.0 months; range: 5.8-34.0). Ninenty-six percent of the patients were treated according to the COBRA guidelines. CONCLUSION: No malignancies were missed during the limited follow-up period, and therefore, to date, the sensitivity of SLCNB seems to be comparable to that estimated in the study setting. PMID- 12756879 TI - [Behcet's disease in a Dutch boy with painful skin lesions]. AB - A 14-years-old Dutch boy had pain in both calves, recurrent oral ulcers, fever, hoarseness and erythema nodosum-like skin lesions. Laryngoscopic examination revealed a vocal cord ulcer and leukocytoclastic vasculitis was visible in a skin biopsy. Based on the clinical picture, the diagnosis of 'incomplete juvenile Behcet's disease' was established. During treatment with colchicine, genital ulcers developed. These disappeared after local treatment with corticosteroids. Juvenile Behcet's disease is rare in Western Europe and the diagnosis is often difficult. Diagnostic criteria have been formulated, but in most cases there is a delay before these criteria are fulfilled due to the slow clinical course of the disease. All organ systems may be affected; mucocutaneous and skin lesions are the most frequent manifestations. Therapy depends on the severity and the symptoms of the disease. PMID- 12756880 TI - [Carcinoma of the fallopian tube after prophylactic laparoscopic ovariectomy in a patient with a BRCAI mutation]. AB - A 48-year-old woman with a distended abdomen appeared to have ascites and was admitted to the gynaecological ward. At the age of 31 years she had been diagnosed with breast cancer and had undergone surgical breast conservation of the right breast. There was a history of both ovarian cancer and breast cancer in her family. Genetic evaluation showed that she was carrying a BRCAI germline mutation. At the age of 42 years she underwent a prophylactic bilateral laparoscopic ovariectomy and 5 years later she underwent a complete mastectomy due to breast carcinoma of the left breast. Two months later she developed ascites, a raised CA125 level and on a CT scan carcinoma of the peritoneum. During the laparotomy a fallopian tube carcinoma was found. After the uterus, fallopian tubes and omentum had been surgically removed, chemotherapy took place. The patient tolerated this well and the CA125 value decreased. Recently, the first molecular evidence was found that linked fallopian tube cancer to germline mutations in BRCAI patients. Patients harbouring a BRCA germline mutation not only have an increased risk of ovarian carcinoma but also of fallopian tube carcinoma. Therefore, in patients with a BRCA mutation, prophylactic surgery should take the form of an adnexectomy, not an oophorectomy. PMID- 12756881 TI - [Diagnostic image (129). A woman who collapsed upon catheterisation]. PMID- 12756882 TI - [Mechanical ventilation in acute respiratory distress syndrome (ARDS): lung protecting strategies for improved alveolar recruitment]. PMID- 12756883 TI - Exporting occupational disease. PMID- 12756884 TI - A short history of pathology registries, with emphasis on cancer registries. AB - In the 1950s, major technical problems for ensuring quality and effectiveness of population based registration of cancer and other conditions had been faced and solved. Nevertheless, the classical epidemiological texts published in the 1960s and the 1970s gave little attention to and showed limited enthusiasm for population-based registries of pathology. The latter, in fact, have been marginal to the dramatic evolution of basic conceptual issues such as study design and causal inference. This perspective may change with the use of registries in order to assess quality of care in a public health perspective. In looking retrospectively, marked changes occurred since the 1960s in distribution between countries and continents of cancer registries to the dataset known as "Cancer Incidence in Five Continents" (where worldwide data from cancer registries of quality converge). The number of countries with cancer registries approximately doubled vs. a fivefold increase in both the number of active cancer registries and the total population served by registration. Nevertheless the increases were concentrated in developed countries whereas in developing countries there was a substantial decrease in registration, attributable to the political and economical situation. PMID- 12756885 TI - [Shared decision making: an overview of international research literature]. AB - Shared decision making (SDM) is a particular type of physician-patient interaction to which the patient brings his/her individual preferences and the physician contributes the medical expertise. The aim of the SDM process is a treatment decision based on mutual agreement and active participation. This kind of decision-making is best effectuated in cases of diseases with medical uncertainty and/or differing patient outcome-treatment potentialities (e.g., breast- or prostate cancer). The concept of SDM is widely known in the English speaking world. A database search with the keywords "shared decision" produced 301 relevant papers, of which 193 are theoretical and 108 empirical works. This paper gives an overview over the state of international research under special consideration in continental European literature. Different questions are explored with regard to potential methodological and contextual research fields. Present results indicate that the level of patient preference to participate in decisions is higher than their actual involvement. Results also prove that SDM leads to higher rates of satisfaction and better treatment results according to patients. Results regarding the efficacy of various intervention methods used to promote SDM, especially within different cultural contexts, are inconsistent. A great amount of research still needs to be done in this field. PMID- 12756886 TI - Prevention of neural tube defects: effect of an intervention aimed at implementing the official recommendations. AB - OBJECTIVES: The periconceptional intake of 0.4 mg folic acid is recommended in many countries to prevent neural tube defects. This paper describes the poor implementation of corresponding guidelines in Germany, the effectiveness of an intervention-based on providing adequate information and the problems associated with the implementation. METHODS: Two cross sectional studies investigated knowledge, attitude, and behaviour of randomly sampled gynaecologists (n = 24/27), pharmacists (n = 21/21), and women in childbed (n = 131/118) before and after the information campaign. RESULTS: Before the intervention, 3.8% of the women implemented folic-acid-prophylaxis compared with 9.3% afterwards (p = n.s.). The awareness of the prophylaxis before pregnancy correlated with socio economic status and rose from 28% (before) to 42% after intervention (p < 0.05). Before the intervention, 38% of the gynaecologists and 38% of the pharmacists recommended the prophylaxis compared with 74% (p < 0.05) and 43% (p = n.s.) afterwards. CONCLUSIONS: The effect of the intervention was small. To improve the situation, fortification of specially selected and labelled food should be considered. An accompanying nationwide information campaign should focus on women with lower socio-economic status. PMID- 12756888 TI - [Educational status and occupational training, occupational status and ischemic heart diseases: a prospective study with data from statutory health insurance in Germany]. AB - OBJECTIVES: To study associations between education, occupational position, and incidence of ischemic heart disease (ICD-9 410-414). METHODS: A cohort of 151,471 male and female members of a German statutory health insurance company aged between 25 and 65 years was investigated. The cohort comprised all members between 1987 and 1996. Information on ischemic heart disease was derived from clinical diagnosis. Education, training, and occupational position according to the British Registrar General defined the indicators of social status. RESULTS: After adjustment for age and length of observation period, education and training as well as occupational position were associated with the incidence of ischemic heart disease in both men and women. Whereas a gradient was observed in men regarding education and training (odds ratios (OR): 3.41-6.02) men with lower occupational position had higher risk estimates as compared to the highest occupational status group (OR: 1.73-3.05). Among women a gradient was observed concerning education and training (OR: 1.75-3.78). With regard to occupational status position female members of the highest group showed the lowest risk as compared to the lower status groups (OR: 1.58-2.19). CONCLUSIONS: Social inequality in ischemic heart disease morbidity was observed among male and female members of a German statutory health insurance. Findings are of importance for health policy and call for preventive action. PMID- 12756887 TI - The Victorian ambulatory care sensitive conditions study: rural and urban perspectives. AB - OBJECTIVES: Ambulatory Care Sensitive Conditions (ACSCs) are those for which hospitalisation is thought to be avoidable with the application of preventive care and early disease management, usually delivered in the ambulatory setting. This study presents detailed analyses of ACSCs as a measure of health outcome that might vary with access to primary health care in rural and urban regions of Victoria. METHOD: The Victorian Admitted Episodes Dataset (VAED), and data from the Health Insurance Commission, Medical Labour Force Annual Survey, socio economic indexes for areas, and accessibility/remoteness index of Australia were merged to identify individual and aggregate level predictors of urban/rural differentials of ACSCs. Estimates of odds ratios and 95% confidence intervals were based on random effect multi-level generalised linear models. RESULTS: After adjustment for age, sex, and severity of illness, significant predictors of higher admission rates of ACSCs within rural areas include lack of insurance, emergency admissions, higher degree of remoteness, lower population density, lower number of general practitioners/10,000 population by local government area (LGA), lower number of general practitioner visits per person by LGA, and areas with lower socio-economic status, education and occupation, and economic resources. CONCLUSIONS: This study suggests that lack of timely and effective care may have a significant impact on rates of admissions for ACSCs in rural areas of Victoria especially in lower socio-economic groups. PMID- 12756889 TI - [Questions about questions: cognitive survey of questionnaire development]. AB - OBJECTIVES: Traditional pretests are used in the development of survey items to identify technical and comprehension problems. Cognitive processes involved in answering survey questions are not the object of this kind of test. METHODS: Cognitive survey methods were used here to test a questionnaire screening for rehabilitation needs in people suffering from back pain. Essential techniques of cognitive testing (think-aloud, probing, confidence ratings) are outlined. We applied these techniques to 20 patients suffering from either acute or chronic back pain in order to test the survey. RESULTS: The main goal, i.e., identifying problems in item formulation by means of cognitive testing, was achieved. Almost one third of the survey questions were rephrased according to the results of the study. Some of the improvements of the questionnaire are illustrated. CONCLUSIONS: The increased effort required to perform cognitive testing as compared to traditional pretesting pays off. The two methods have specific pros and cons and cannot replace one another. PMID- 12756890 TI - Mesotheliomas among Italians, returned to the home country, who worked when migrant at a cement-asbestos factory in Switzerland. AB - OBJECTIVES: To report the occurrence of mesotheliomas in Italy among subjects who worked, when migrant, at a cement-asbestos factory in Niederurnen, Switzerland, and had resettled to the home country. METHODS: Information about the disease and on the working history of subjects was collected by regional mesothelioma registries. Only cases diagnosed by means of histo-pathological examinations have been considered here. RESULTS: 15 mesotheliomas (13 pleural, 2 perithoneal; 12 among males, 3 among females) have been identified among Italians, who had worked at the factory. None of them had other occupational exposure to asbestos. The majority was living in the Veneto Region (North East of Italy), and in Puglia (Southern Italy). CONCLUSIONS: Exposure to asbestos at this factory has already caused an important number of occupational cancers among the employees, a large fraction being constituted of migrants. In order to avoid under-estimation of risks and to allow compensation, diseases which occur among foreign workers returned to their home country should be evaluated. Migration for work is at the genesis of asbestos-related mesotheliomas now occurring in Italy. PMID- 12756891 TI - Asking sensitive information: an example with income. PMID- 12756893 TI - Apoptotic detection methods--from morphology to gene. AB - To date, many terms have been given for cell death and forgotten before they became widely accepted. Most researchers studying cell death use some terms for cell death that have survived over centuries and regard apoptosis and programmed cell death (PCD), and necrosis and oncosis as synonymous. The different terminologies used for cell death depending on the preference of researchers sometimes cause confusion in the study of apoptosis. The study of apoptosis was first based on cell morphology using transmission electron microscopy (TEM): chromatin condensation, cellular shrinkage, budding and apoptotic body formation. Recently, marked progress in biochemistry, molecular biology and genetics provided researchers of apoptosis various tools for apoptosis detection, such as the terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick end labeling (TUNEL) method, agarose gel electrophoresis using extracted DNA, staining methods using fluorescence dyes, and flow cytometry. This review focuses on the following topics: history of the study of cell death and its classification, apoptosis-related proteins and their signal pathways, morphological, biochemical and molecular biological methods of apoptosis detection, and the transcriptional regulation of bcl-2 using the real-time Southwestern method and cancer gene therapy. In addition, the merits and demerits of the above-mentioned apoptosis detection methods are discussed. PMID- 12756892 TI - The diffuse endocrine system: from embryogenesis to carcinogenesis. AB - In the present review we will summarise the current knowledge about the cells comprising the Diffuse Endocrine System (DES) in mammalian organs. We will describe the morphological, histochemical and functional traits of these cells in three major systems gastrointestinal, respiratory and prostatic. We will also focus on some aspects of their ontogeny and differentiation, as well as to their relevance in carcinogenesis, especially in neuroendocrine tumors. The first chapter describes the characteristics of DES cells and some of their specific biological and biochemical traits. The second chapter deals with DES in the gastrointestinal organs, with special reference to the new data on the differentiation mechanisms that leads to the appearance of endocrine cells from an undifferentiated stem cell. The third chapter is devoted to DES of the respiratory system and some aspects of its biological role, both, during development and adulthood. Neuroendocrine hyperplasia and neuroendocrine lung tumors are also addressed. Finally, the last chapter deals with the prostatic DES, discussing its probable functional role and its relevance in hormone resistant prostatic carcinomas. PMID- 12756894 TI - [Socio-economic repercussion of rheumatoid arthritis]. PMID- 12756895 TI - [Approximation to the cost of pharmacological treatment of rheumatoid arthritis in Spain]. AB - OBJECTIVE: To study the direct cost derived of utilization of pharmaceutical compounds in the treatment of rheumatoid arthritis(RA). MATERIAL AND METHODS: A prospective study with 150 patients (125 women/25 men) suffered of RA was carry out in Galicia-Spain public hospital. The mean age was 60.2 years, with a mean lengthy of disease of 11.2 years (1-53); the 64.4% come from rural areas. A personal interview was made with a complete registry of all data: demographic, activity score (ESR, Swollen joints), radiological status, functional class (ACR), extrarticular manifestations and co-morbid diseases. Such data was accomplished with all the medications employees with a calculation of monthly or annual cost for the different therapeutic groups and a final total cost. The statistical study was made with Excel's (Microsoft) and the Analysis Tool Pack. RESULTS: The 53% of patients was in functional class I and 52% in radiological stage I or II (Steinbrocker) whereas 16 patients was considered in remission. Non steroideal ant-inflammatory drugs (NSAID) were used in the 82.7% of patients with a monthly cost of 12.71 [symbol: see text] (1.10-80). Corticosteroids at low doses were used in 90.7% with a monthly cost of 5.17 [symbol: see text] (1.24 33.7). The Disease Modifying Anti-rheumatic Drugs (DMARD) was used in 94% of cases, the most common methotrexate and association of two or more in 21%. The mean monthly cost for a single DMARD was 3.63 [symbol: see text] and for two, 13.75 [symbol: see text]. Gastroprotection and therapy for co-morbid diseases was employee in 80% and 95% of cases, with a monthly cost of 36.9 [symbol: see text] and year cost of 568.6 [symbol: see text], respectively. The study included 23 patients under treatment with anti-TNF therapy with a monthly mean cost of 933.8 [symbol: see text]. For pharmaceuticals exclusively for RA, annual cost was 342.8 [symbol: see text] excluding anti-TNF therapy, but with wide variation (6.4-2.910 [symbol: see text]). If we include all patients with anti-TNF therapy, gastro protection and co-morbid situations in a calculation, the mean cost was 2.587 [symbol: see text] year. The most important cost was found in patients with 50-70 years-old and existing a good correlation between the final burden and use of medications for co-morbid conditions, gastro-protection, use of anti-TNF, age, lengthy of disease between 5-10 years and number of swollen joints, but not radiological stage. CONCLUSIONS: The economic burden for pharmaceuticals used in RA is very variable depending of some variables, including the proper disease and other related conditions. The most important cost occur in case of use of anti TNF therapy. In the most frequent conditions, gastro-protection and therapy for co-morbid diseases lead the 62% of total annual burden, followed by the use of DMARD and in a minor load, the NSAIDs and corticosteroids. PMID- 12756896 TI - [Glycemic control related factors in type 2 diabetes mellitus patients]. AB - OBJECTIVES: To describe metabolic, lipidic and blood pressure control and to identify blood glucose control predictive factors in patients with type 2 diabetes. MATERIAL AND METHODS: Retrospective cohort study. Ambulatory type 2 diabetic patients were selected from 2/1/00 to 14/11/01 if they had been visited yearly at least three years previously. They were classified into three groups: a) initial and final treatment with diet and/or oral glucose-lowering drugs; b) insulinized patients during follow-up; and c) initial and final therapy with insulin. BODY MASS INDEX: (BMI), blood pressure, HbA1c and lipid profile were determined at start and at the end of the study. A multiple regression model was used to predict mean HbA1c through follow-up. RESULTS: 320 patients were selected, 199 women (62.2%) and 121 men (37.8%) with mean age of 69 years (SD: 9). Mean follow-up was 5.2 years (SD: 1.1). 106 patients (33.8%) were included into A group, 70 patients (22.4%) into B group and 137 (43.8%) into C group. 63% of patients had values of HbA1c > 7.5%, 36% blood pressure > 140/85 mmHg and 8.5% LDL values > 155 mg/dl. HbA1c mean through follow-up was of 8.2% and its independent predictors were baseline HbA1c (0.37; IC 95% 0.31-0.43; p < 0.001), baseline BMI (0.023; IC 95% 0.004-0.042; p = 0.017), baseline treatment with insulin (0.67; IC 95% 0.45-0.88; p < 0.001) and insulinization during follow-up (0.46; IC 95% 0.21-0.71; p < 0.001). CONCLUSIONS: It's difficult to achieve an acceptable blood glucose control. Mean HbA1c predictive factors include baseline HbA1c and BMI and insulin therapy. PMID- 12756897 TI - [Hypothyroidism: clinical and economical implications in a health area]. AB - OBJECTIVE: To analyze the clinical, biochemical and epidemiological characteristics in patients with hypothyroidism, substitutive dose and annual direct costs, in a health area. MATERIAL AND METHODS: Seventy five patients attended in a health area were included in the review. Data was divided in epidemiological, biochemical, clinics and economics. RESULTS: A 94.7% were females and 5.3% males, with an average age of 52.07 + 16.5 years. The most frequent disease was Thyroiditis of Hashimoto 48% and atrophic thyroiditis 46.6%. Average value of TSH was 17.57 + 40.8 mUI/ml and TPO 629.3 + 1049 UI/ml and Ac TG 532.1 + 1028 UI/ml. A 58.7% of patients had goiter, only evident 2.6%. The average dose of T4 was 81 + 53.5 mg/day. The average number of visits was 2.67 + 1.08. The highest cost was due to specialists visit 160 + 65 euros with a total average cost of 212.9 + 81.5 euros. Levels of TPO was higher in T. Hashimoto, with a higher number of visits and costs. CONCLUSIONS: The most prevalent disease was autoimmune (T Hashimoto and atrophic thyroiditis) with higher levels of TPO in Hashimoto. Total cost is higher in T. Hashimoto. PMID- 12756898 TI - [Treatment of deep vein thrombosis with low molecular weight heparins at home]. AB - Treatment of patients with proximal vein thrombosis with low molecular weight heparins is effective and safe. So it allows "hospital at home" care. Among low molecular weight heparins tinzaparin is given once daily, making the compliance easier. The twenty patients with deep vein thrombosis who were assisted in the "hospital at home" unit of the Clinica Nuestra Senora de la Concepcion in Madrid from December 1999 to December 2000. The mean of age was 71 + 15. The most frequent risk factors were surgery in past three months (19%), known tumour (15%) and previous venous thrombosis (15%). Eighteen patients were treated with tinzaparin. No patients showed adverse effects nor complications. Low molecular weight heparins, specially tinzaparin, are a safe and effective treatment for deep vein thrombosis in our unit. PMID- 12756900 TI - [Hepatic granulocytic sarcoma: an unusual presentation]. AB - A case of granulocytic sarcoma (chloroma) of hepatic localization is presented. It is a extramedullary strange tumour, composed of immature precursors of myeloid cells. Clinically it can show, before, during or after a acute myeloid leukemia, chronic myeloproliferative disorders or myelodysplastic syndromes. Our patient, 81 year-old male, presented a process of important acute jaundice, with negative image technics, what indicated us the intrahepatic origin, negative tumorals markers, negative serology and hepatic biopsy (the piece of greenish coloration is described) what showed a hepatic sinusoides diffuse infiltration by indifferentiation cellularity, with study immuno-histochemical that was positive for the myeloperoxydase, giving a diagnose compatible with hepatic infiltration for acute myeloid leukemia. The patient doesn't present affectation of peripheral blood, and he died for acute hepatic and renal failure after 8 days of entrance. PMID- 12756899 TI - [Diagnosis and localization of hyperparathyroidism by nuclear medicine procedures]. AB - Primary hyperparathryoidism is a PTH hypersecretion caused by the parathyroid glands. In most cases (85%), the origin is to be due to the existence of a parathyroid adenoma, despite the intrinsic difficulty in being localized under certain circumstances. From some time now, we can count with the invaluable help of a nuclear medicine technique, namely the parathyroid scintigraphy with Technetium 99m-sestamibi (Tc99m-MIBI), a technique which is easy to perform, cheap and with excellent results, and which additionally can provide us with the above mentioned necessary information regarding location. We present here the case of a patient suffering from primary hyperparatyiroidism, in whom both the disease and the precise location of the hyperfunctioning tissue were identified by means of the parathyroid scintigraphy. Another nuclear medicine procedure, the one known as bone scintigraphy, also contributed meaningfully to the correct diagnosis in the same patient. PMID- 12756901 TI - [Leishmaniasis in a patient with liver cirrhosis]. AB - We present a 35 years-old man with fever related leishmania infection as the first complication of a previously undiagnosed cirrhosis. Diagnosis of leishmaniasis is often complicated. Clinical and therapeutical aspects are commented. PMID- 12756902 TI - [Treatment of chronic obstructive pulmonary disease]. AB - Chronic obstructive pulmonary disease (COPD) is a highly prevalent condition and most epidemiological studies have found that COPD prevalence, morbidity and mortality have increased over the last few years. Recent trials suggest that treatments are improving. Several studies have demonstrated the usefulness of the long-acting inhaled beta-2-agonists. Inhaled glucocorticoids, although unable to alter the progression of COPD, may reduce the frequency of acute exacerbations and health status deterioration in a limited group of patients with COPD. Likewise, these drugs may offer benefits in combination with long-acting inhaled beta-agonists. Finally, a better understanding of the molecular and cellular mechanisms involved in COPD pathogenesis should lead to effective treatments that slow or halt the course of the disease. New classes of agents such as the phosphodiesterase inhibitors are now in development. PMID- 12756903 TI - [Anaphylaxis as first manifestation of unknown hydatid disease]. PMID- 12756904 TI - [A new case of Creutzfeld-Jacob disease]. PMID- 12756905 TI - [Secondary plasma cell leukemia as evolution of monoclonal gammapathy]. PMID- 12756906 TI - [Erysipelothrix rhusiopathiae endocarditis, a new case report]. PMID- 12756907 TI - [Aortic dissection, report of two cases]. PMID- 12756908 TI - [Primary cholelithiasis in a situs inversus totalis patient]. PMID- 12756910 TI - [Lung abscess caused by Prevotella oralis and Prevotella ruminicola in a HIV positive patient]. PMID- 12756909 TI - [Upper airway obstruction by angioedema following thrombolytic therapy with recombinant tissue plasminogen activator]. PMID- 12756911 TI - [Low height and mental retardation]. PMID- 12756912 TI - Purification and characterization of a thermostable beta-galactosidase from kidney beans (Phaseolus vulgaris L.) cv. PDR14. AB - Using five different steps, beta-Galactosidase has been purified from kidney beans to apparent electrophoretic homogeneity with approximately 90-fold purification with a specific activity of 281 units mg-1 protein. A single band was observed in native PAGE. Activity staining of the native gel with 5-bromo 4 chloro 3-indoxyl beta-D-galactopyranoside (X-Gal) at pH 4.0 also produced a single band. Analytical gel filtration in Superdex G-75 revealed the molecular mass of the native protein to be approximately 75 kD. 10% SDS-PAGE under reducing conditions showed two subunits of molecular masses, 45 and 30 kD, respectively. Hence, beta-galactosidase from kidney beans is a heterodimer. A typical protein profile with lambda max at 280 nm was observed and A280/A260 ratio was 1.52. The N-terminal sequence of the 45 kD band showed 86% sequence homology with an Arabidopsis thaliana and 85% with Lycopersicon esculentum putative beta galactosidase sequences. The Electrospray Mass Spectrometric analysis of this band also revealed a peptide fragment that had 90% sequence homology with an Arabidopsis thaliana putative beta-galactosidase sequence. The N-terminal sequencing of the 30 kD band as well as mass spectrometric analysis both by MALDI TOF and ES MS revealed certain sequences that matched with phytohemagglutinin of kidney beans. The optimum pH of the enzyme was 4.0 and it hydrolysed o- and p nitrophenyl beta-D galactopyranoside with a Km value of 0.63 mmol/L and 0.74 mmol/L, respectively. The energy of activation calculated from the Arrhenius equation was 14.8 kcal/mol enzyme site. The enzyme was found to be comparatively thermostable showing maximum activity at 67 degrees C. Thermal denaturation of the enzyme at 65 degrees C obeys single exponential decay with first order-rate constant 0.105 min-1. Galactose, a hydrolytic product of this enzyme was a competitive inhibitor with a Ki of 2.7 mmol/L. PMID- 12756913 TI - Putrescine facilitated enhancement of capsaicin production in cell suspension cultures of Capsicum frutescens. AB - Putrescine treatment (0.1 mmol/L) influenced enhancement of growth and capsaicin production in the cell suspension cultures of C. frutescens. The administration of polyamine inhibitor DFMA (alpha-DL-difluoromethylarginine) resulted in a reduction of the growth, capsaicin content and the endogenous titres of polyamines (PAs). The capsaicin synthase activity was also higher in the putrescine (Put) treated cultures. Ethylene levels were lower in the cultures treated with putrescine. This study suggested that Put facilitates growth and capsaicin production. PMID- 12756914 TI - Enhanced stress-tolerance of transgenic tobacco plants expressing a human dehydroascorbate reductase gene. AB - To analyze the physiological role of dehydroascorbate reductase (DHAR, EC 1.8.5.1) catalyzing the reduction of DHA to ascorbate in environmental stress adaptation, T1 transgenic tobacco (Nicotiana tabacum cv. Xanthi) plants expressing a human DHAR gene in chloroplasts were biochemically characterized and tested for responses to various stresses. Fully expanded leaves of transgenic plants had about 2.29 times higher DHAR activity (units/g fresh wt) than non transgenic (NT) plants. Interestingly, transgenic plants also showed a 1.43 times higher glutathione reductase activity than NT plants. As a result, the ratio of AsA/DHA was changed from 0.21 to 0.48, even though total ascorbate content was not significantly changed. When tobacco leaf discs were subjected to methyl viologen (MV) at 5 mumol/L and hydrogen peroxide (H2O2) at 200 mmol/L, transgenic plants showed about a 40% and 25% reduction in membrane damage relative to NT plants, respectively. Furthermore, transgenic seedlings showed enhanced tolerance to low temperature (15 degrees C) and NaCl (100 mmol/L) compared to NT plants. These results suggest that a human derived DHAR properly works for the protection against oxidative stress in plants. PMID- 12756915 TI - Seasonal changes in oleosomic lipids and fatty acids of perennial root nodules of beach pea. AB - Seasonal changes in the fatty acid composition of phospholipids (PL), monoglycerides (MG), diglycerides (DG), free fatty acids (FA) and triglycerides (TG) separated from oleosomes (lipid bodies) of perennial root nodules of beach pea (Lathyrus maritimus) were analysed. Thin layer chromatography (TLC) revealed that PL and MG are the major lipids in nodule oleosomes. The fatty acid profile and overall double bond index (DBI) varied among lipid classes depending upon the season. High DBI in PL and MG found during late winter and early spring indicated that they may play a major role in winter survival and regeneration of perennial nodules. The DBI of DG was high at the end of the fall season and the DBI of FA and TG was high in summer months. The dominant fatty acids are C16:0 followed by C18:0 and C18:1. The levels of many unsaturated fatty acids such as C18:1, C18:2 and C18:3 increased while saturated fatty acid C18:0 decreased during winter. These unsaturated fatty acids possibly play an important role in the protection of nodule cells from cold stress. Nodules seem to retain some fatty acids and selectively utilize specific fatty acids to survive the winter and regenerate in spring. PMID- 12756916 TI - Effects of cadmium on the co-ordination of nitrogen and carbon metabolism in bean seedlings. AB - The effect of cadmium (Cd) was investigated on the in vitro activities of leaf and root enzymes involved in carbon (C) and nitrogen (N) metabolism of bean (Phaseolus vulgaris L. cv. Morgane). Cd induced a high increase in maximal extractable activity of glutamate dehydrogenase (NADH-GDH, EC 1.4.1.2). Cd promoted ammonium accumulation in leaves and roots, and a tight correlation was observed between ammonium amount and GDH activity. Changes in GDH activity appear to be mediated by the increase in ammonium levels by Cd treatment. Cd stress also enhanced the activities of phosphoenolypyruvate carboxylase (PEPC, EC 4.1.1.31) and NADP(+)-isocitrate dehydrogenase (NADP(+)-ICDH, EC 1.1.1.42) in leaves while they were inhibited in roots. Immuno-titration, the PEPC sensitivity to malate and PEPC response to pH indicated that the increase in PEPC activity by Cd was due to de novo synthesis of the enzyme polypeptide and also modification of the phosphorylation state of the enzyme. Cd may have modified, via a modulation of PEPC activity, the C flow towards the amino acid biosynthesis. In leaves, Cd treatments markedly modified specific amino acid contents. Glutamate and proline significantly accumulated compared to those of the control plants. This study suggests that Cd stress is a part of the syndrome of metal toxicity, and that a readjustment of the co-ordination between N and C metabolism via the modulation of GDH, PEPC and ICDH activities avoided the accumulation of toxic levels of ammonium. PMID- 12756918 TI - In vivo and in vitro effects of cadmium on H+ ATPase activity of plasma membrane vesicles from oat (Avena sativa L.) roots. AB - The effect of an in vivo and in vitro treatment with cadmium on transport activities of root plasma membrane enriched vesicles was studied in oat (Avena sativa L. cv. Argentina) plants. Addition of 100 mumol/L CdSO4 to nutrient solution decreases both proton transport activity and ATPase activity to the same level. In vitro experiments show that cadmium seems to have a differential inhibiting effect on proton transport activity and ATPase activity, the most pronounced one on ATP-dependent H(+)-accumulation, suggesting that cadmium would interfere with membrane permeability properties. This is indeed the case. The results demonstrate that cadmium decreases passive permeability to protons. PMID- 12756917 TI - Nitrogen fixation in transposon mutants from Bradyrhizobium japonicum USDA 110 impaired in nitrate reductase. AB - Tn5 transposon mutagenesis was carried out in Bradyrhizobium japonicum strain USDA 110 to produce defective mutants. From over one thousand clones expressing low levels of nitrate reductase activity as free-living bacteria, approximately five percent had significantly different ratios of nodulation, N2 fixation or nitrate reductase activity compared to the wild strain when determined in bacteroids from soybean nodules. Tn5 insertions were checked previously and mutants were arranged into four different groups. Only one of these groups, designated AN, was less effective at N2 fixation than the wild strain, suggesting a mutation in a domain shared by nitrogenase and NR. The remaining groups of insertions successfully nodulated and were as effective at N2 fixation as the wild strain, but showed diminished ability to reduce nitrate both in nodules and in the isolated bacteroids when assayed in vitro with NADH or methyl viologen as electron donors. PCR amplification demonstrated that Tn5 insertions took place in different genes on each mutant group and the type of mutant (CC) expressing almost no nitrate reductase activity under all treatments seemed to possess transposable elements in two genes. Induction of nitrate reductase activity by nitrate was observed only in those clones expressing a low constitutive activity (AN and AE). Nitrate reductase activity in bacteroids along nodule growth decreased in all groups including the ineffective AN group, whose nodulation was highly inhibited by nitrate at 5 mmol/L N. Host-cultivar interaction seemed to influence the regulation of nitrate reductase activity in bacteroids. Total or partial repression of nitrate reductase activity in bacteroids unaffected by N2 fixation (CC, AJ and AE groups) improved nodule resistance to nitrate and N yields of shoots over those of the wild strain. These observations may suggest that some of the energy supplied to bacteroids was wasted by its constitutive NRA. PMID- 12756919 TI - Seasonal changes in the levels of compatible osmolytes in three halophytic species of inland saline vegetation in Hungary. AB - Seasonal changes in the leaf concentration of compatible osmolytes were investigated in three halophytic species (Lepidium crassifolium, Camphorosma annua and Limonium gmelini subsp. hungaricum) native to a salty-sodic grassland. The investigated species were shown to accumulate both carbohydrate- and amino acid-derived osmolytes. The leaf tissues of C. annua (Chenopodiaceae) preferentially stored glycine betaine and pinitol, while in L. gmelini (Plumbaginaceae) beta-alanine betaine, choline-O-sulphate, and pinitol were accumulated. In the leaves of L. crassifolium (Brassicaceae) a very high amount of proline, associated with a high level of soluble carbohydrates was found. Not only the biochemical nature of the osmolyte, but also the seasonal pattern of osmolyte accumulation showed significant species-specific fluctuations. In addition, the cellular levels of the observed osmolytes changed with the growth period and according to the environmental parameters. The highest concentrations of osmolytes were found in March, when low temperatures, hypoxic conditions and high salt concentrations were the main constraints to plant growth. The high structural diversity of osmolytes combined with their multifunctionality and the seasonal flexibility of the metabolism in plants facing multiple stresses is discussed. PMID- 12756920 TI - Photosystem II photochemistry and photosynthetic pigment composition in salt adapted halophyte Artimisia anethifolia grown under outdoor conditions. AB - The effects of high salinity (0-400 mmol/L NaCl) on photosystem II (PSII) photochemistry and photosynthetic pigment composition were investigated in the halophyte Artimisia anethifolia grown under outdoor conditions and exposed to full sunlight. High salinity resulted in an inhibition in plant growth and a significant accumulation of sodium and chloride in leaves. However, high salinity induced no effects on the actual PSII efficiency, the efficiency of excitation energy capture by open PSII reaction centres, photochemical quenching, and non photochemical quenching at midday. High salinity also induced neither changes in the maximum efficiency of PSII photochemistry, the efficiency with which a trapped exciton can move an electron into the electron transport chain further than QA and the quantum yield of electron transport beyond QA, nor changes in absorption, trapping and electron transport fluxes per PSII reaction centre. No significant changes were observed in the levels of neoxanthin, lutein, beta carotene, violaxanthin, antheraxanthin, and zeaxanthin expressed on a total chlorophyll basis in salt-adapted plants. Our results suggest that Artimisia anethifolia showed high resistance not only to high salinity, but also to photoinhibition even if it was treated with high salinity as high as 400 mmol/L NaCl and exposed to full sunlight. The results indicate that tolerance of PSII to high salinity and photoinhibition can be viewed as an important strategy for Artimisia anethifolia, a halophyte plant, to grow in very high saline soil. PMID- 12756921 TI - Development of the primary root and mobilisation of reserves in etiolated seedlings of Brassica napus grown on a slowly rotating clinostat. AB - The effect of the slow rotating clinostat (1 rpm) on the growth of the primary root was studied on Brassica napus seedlings. After 5 d in darkness, the primary root was longer and thinner in seedlings grown on the clinostat than in seedlings grown in the vertical position. However, the breakdown of lipid reserves, sucrose level and transport of 14C-labeled sucrose from the cotyledons to the primary root, were not altered by growth on the clinostat. Moreover, the activity of isocitrate lyase, one of the two enzymes necessary for the conversion of lipids into glucids also was also not modified in the cotyledons of clinorotated seedlings. Thus, there was clear evidence that clinorotation had a direct effect on the growth of the primary root that was independent of the mobilisation of lipid reserves in the cotyledons. As a sink, the primary root had the same strength on the clinostat as in the vertical position, but the reserves were used in a different way. The increase in root elongation on the clinostat could be due to the slight, but continuous, omnilateral gravitropic stimulation due to the rotation of the seedlings about a horizontal axis. PMID- 12756922 TI - Somatic embryogenesis and plant regeneration from cotyledon explants of a timber yielding leguminous tree, Dalbergia sissoo Roxb. AB - Efficient plant regeneration through somatic embryogenesis was achieved from callus cultures derived from semi-mature cotyledon explants of Dalbergia sissoo Roxb., a timber-yielding leguminous tree. Somatic embryos developed over the surface of embryogenic callus and occasionally, directly from cotyledon explants without intervening callus phase. Callus cultures were initiated from cotyledon pieces of D. sissoo on Murashige and Skoog (1962) medium supplemented with 4.52, 9.04, 13.57, and 18.09 mumol/L 2,4-dichlorophenoxyacetic acid and 0.46 mumol/L Kinetin. Maximum percentage response for callus formation was 89% on MS medium supplemented with 9.04 mumol/L 2,4-D' and 0.46 mumol/L Kn. Somatic embryogenesis was achieved after transfer of embryogenic callus clumps to 1/2-MS medium without plant growth regulators (1/2-MSO). Average numbers of somatic embryos per callus clump was 26.5 on 1/2-MSO medium after 15 weeks of culture. Addition of 0.68 mmol/L L-glutamine to 1/2-MSO medium enhanced somatic embryogenesis frequency from 55% to 66% and the number of somatic embryos per callus clump from 26.5 to 31.1. Histological studies were carried out to observe various developmental stages of somatic embryos. About 50% of somatic embryos converted into plantlets on 1/2-MSO medium containing 2% sucrose, after 20 days of culture. Transfer of somatic embryos to 1/29-MSO medium containing 10% sucrose for 15 days prior to transfer on 1/2-MS medium with 2% sucrose enhanced the conversion of somatic embryos into plantlets from 50 to 75%. The plantlets with shoots and roots were transferred to 1/2 and 1/4-liquid MS medium, each for 10 days, and then to plastic pots containing autoclaved peat moss and compost mixture (1:1). 70% of the plantiets survived after 10 weeks of transfer to pots. 120 regenerated plantlets out of 150 were successfully acclimatised. After successful acclimatisation, plants were transferred to earthen pots. PMID- 12756923 TI - Silver nitrate and aminoethoxyvinylglycine promote in vitro adventitious shoot regeneration of pomegranate (Punica granatum L.). AB - A protocol is presented for direct adventitous shoot organogenesis and complete plant regeneration from seedling-derived explants of pomegranate (Punica granatum L.), a tropical fruit tree. Murashige and Skoog (1962) (MS) medium enriched with 8.9 mumol/L benzyladenine (BA), 5.4 mumol/L naphthaleneacetic acid (NAA) and 10% coconut water (CW) induced adventitious shoot bud differentiation in axenic seedling-derived cotyledons as well as hypocotyl segments. The cotyledons were more responsive than the hypocotyls. Addition of ethylene inhibitors such as AgNO3 (10-40 mumol/L) and aminoethoxyvinylglycine (AVG) (5-15 mumol/L) to the medium markedly enhanced regeneration frequency as well as number of shoots obtained per explant. The promotive effect of AVG and AgNO3 on shoot organogenesis was observed only in cotyledon explants. The regeneration medium containing AgNO3 (20 mumol/L) or AVG (10 mumol/L) induced adventitious shoot buds from 57% or 53% of the cotyledon explants respectively. These shoot buds developed into shoots upon transfer to a regeneration medium without AgNO3 and AVG. The promotive effect of AVG on shoot regeneration was reversed by exogenous application of 20 mumol/L 2-chloroethylphosphonic acid (CEPA), an ethylene releasing compound. On the other hand, shoot regeneration stimulated by AgNO3 was relatively less affected by CEPA. Regenerated shoots were rooted in half-strength MS medium (1/2 MS) containing 0.54 mumol/L NAA. The well rooted plantlets were acclimatized and eventually established in soil. PMID- 12756924 TI - Alternate energy dissipation? Phenolic metabolites and the xanthophyll cycle. AB - The dynamics of phenolic galloylglucoses (di-, tri-, tetra- and penta galloylglucose), flavonoids (quercitin and quercitin glycosides) and sideroxylonal were compared with that of xanthophyll cycle-dependent energy dissipation during rapid induction of chilling-dependent photo-inhibition. Pre dawn xanthophyll cycle engagement of seedlings of Eucalyptus nitens transferred from mild nursery conditions to a low temperature controlled environment increased logarithmically during eight days of treatment. Photochemical efficiency and flavonoids decreased after four days of treatment and non photochemical quenching after two days of treatment. Galloylglucoses and sideroxylonal decreased linearly during treatment. These results demonstrate that rapid changes in foliar phenolic levels are associated with abrupt changes in the plant environment. It is argued that under these growth-chamber conditions, the xanthophyll cycle facilitated dissipation of excess light energy, lessening the requirement for the dissipation of energy or antioxidant activity through phenolic metabolites. PMID- 12756925 TI - Impact--a new "high" for the Journal of Orofacial Pain. PMID- 12756926 TI - Longitudinal outcome of temporomandibular disorders: a 5-year epidemiologic study of muscle disorders defined by research diagnostic criteria for temporomandibular disorders. AB - AIMS: To investigate the course of myofascial pain defined by Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD) over a period of 5 years, and to identify prognostic factors from baseline data. METHODS: Subjects were 155 consecutive patients and 80 community cases identified from an age-stratified representative population sample; all met the primary selection criterion of reporting pain in the temporomandibular joint (TMJ) or masticatory muscles. The 2 groups were combined to yield a total sample of 235 subjects (50 male, 185 female; mean age = 39 years). Subjects were evaluated at baseline, 1 year, 3 years, and 5 years by trained examiners using standardized, reliable methods. Psychological and behavioral factors were assessed by self report. RESULTS: According to RDC/TMD criteria, 50 (31%) of the 165 subjects presenting with myofascial pain (MFP) at baseline continued to have their disorder over a period of 5 years; 55 (33%) remitted, and 60 (36%) were recurrent cases. Bivariate statistics and multivariate logistic regression analyses indicated that baseline pain frequency, number of painful palpation sites, and total number of body sites with pain were significant predictors of persistent vs remitted and recurrent cases. No predictors that distinguished remission vs recurrence could be identified. Thirty subjects from the 70 without a diagnosis of MFP at baseline developed a new MFP. A high baseline somatization score (without pain items) was a significant risk factor for onset of MFP. CONCLUSION: Muscle disorders classified by RDC/TMD are predominantly chronic or fluctuating pain conditions, with a modest probability (31%) of remission. PMID- 12756927 TI - Prevalence of temporomandibular disorder subtypes, psychologic distress, and psychosocial dysfunction in Asian patients. AB - AIMS: To use the Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD) to investigate the physical diagnoses, psychologic distress, and psychosocial dysfunction in Asian TMD patients. The RDC/TMD Axis I and II findings were compared to those of Swedish and American TMD patients. METHODS: One hundred ninety-one patients (53 male and 138 female) referred to 2 institutionalized TMD clinics in Singapore were enrolled in the study. The mean age of the predominantly Chinese population (83.2%) was 33.6 +/- 9.3 years. Data from a RDC/TMD history questionnaire and clinical examination were fed directly by patients and clinicians into a computerized diagnostic system (NUS TMDv1.1). Axis I and II findings were generated on-line, based on RDC/TMD rule engines. Data were automatically exported to SPSS for statistical analysis. RESULTS: Group I (muscle) disorders were found in 31.4% of the patients; Group II (disc displacement) disorders were found in 15.1% and 15.7% of the patients in the left and right temporomandibular joints, respectively; and Group III (arthralgia, arthritis, and arthrosis) disorders were found in 12.6% and 13.0% of the patients in the left and right joints, respectively. Axis II assessment of psychologic status showed that 39.8% of patients experienced moderate to severe depression and 47.6% had moderate to severe nonspecific physical symptom scores. Psychosocial dysfunction was observed in only 4.2% of patients based on graded chronic pain scores. CONCLUSION: Axis I and II findings of Asian TMD patients were generally similar to their Swedish and American cohorts. In all 3 populations, women of child-bearing age represented the majority of patients. Muscle disorders were the most prevalent type of TMD. A substantial portion of TMD patients were depressed and experienced moderate to severe somatization. PMID- 12756928 TI - Gender difference in symptoms related to temporomandibular disorders in a population of 50-year-old subjects. AB - AIMS: To investigate, by means of a mail questionnaire, the prevalence of symptoms related to temporomandibular disorders (TMD) in 50-year-old subjects living in the counties of Orebro and Ostergotland, Sweden. METHODS: The total population comprised 8,888 individuals, and the overall response rate was 71%. A clinical evaluation of the masticatory system was performed in subgroups to validate the responses to the questionnaire. There was satisfactory correspondence between self-reports and well-defined clinical conditions. RESULTS: Women reported, more often than men, pain from the temporomandibular joints (TMJs), TMJ sounds, bruxism, sensitive teeth, and burning mouth symptoms. The prevalences of difficulties in jaw opening, loss of anterior teeth due to trauma, and masticatory problems were greater in men than in women. No gender difference was found in the number of remaining teeth. Logistic regression analysis with pain from the TMJ as the dependent variable identified bruxism, impaired chewing efficiency, and gender (women) as the most significant risk factors. With reduced chewing ability as the dependent variable, several missing teeth constituted the highest risk, followed by pain from the TMJ, bruxism, gender (men), and loss of anterior teeth due to trauma. CONCLUSION: There were significant gender differences in reported TMD-related symptoms in 50-year-old Swedes. Bruxism was a significant risk factor for pain from the TMJ. Reduced number of teeth and pain from the TMJ were significant risk factors for impaired chewing ability. PMID- 12756929 TI - Complementary and alternative therapy use by patients with myofascial temporomandibular disorders. AB - AIMS: To examine the prevalence and predictors of complementary and alternative medicine (CAM) use among patients with temporomandibular disorders (TMD), prior to their first treatment with an intraoral splint. METHODS: Sixty-three women with a diagnosis of myofascial TMD, and who had never been prescribed an intraoral appliance, reported on their use of CAM and other treatments for their facial pain. In addition to providing a comprehensive symptom history, participants completed a 2-week daily diary in which they described the nature of daily efforts to reduce their facial pain. RESULTS: Although more than half of all participants had not sought any prior treatment for their facial pain, 22.2% had received CAM treatment. The only single type of treatment more commonly used than CAM treatment was medication (28.6%). The most common type of CAM treatment was relaxation therapy (12.7%), followed by chiropractic treatment (9.5%). Although pain duration, pain severity, or mood did not predict CAM use, users were significantly more likely to report work or social disability associated with their facial pain and were more likely to report onset associated with an accident. CAM users were more likely than non-users to employ multiple pain reduction strategies over the 2-week daily diary report, including prescription medication use. CONCLUSION: A sizeable minority of women with myofascial TMD report CAM treatment for their pain, even prior to an initial treatment with an intraoral splint. Since empirical reports have not adequately demonstrated their safety or efficacy, there is a need for controlled clinical trials evaluating the utility of CAM treatments for TMD. PMID- 12756930 TI - Counseling and physical therapy as treatment for myofascial pain of the masticatory system. AB - AIMS: To prospectively evaluate the effectiveness of a treatment regimen comprising counseling and physical therapy in patients with myofascial pain of the masticatory system, and to explore whether the duration of the physical therapy offered (4 vs 6 weeks) would influence the treatment result. METHODS: Twenty-six patients were randomly distributed over 2 groups. All patients received reassuring information, advice regarding relaxation of the jaws, avoiding parafunctions, and limited use of the jaws. In addition, a physical therapy program (heat application, massage, ultrasound and muscle stretching) was initiated 2 weeks after the start of the study (group I, receiving 4 weeks of physical therapy) or immediately from the start of the study (group II, receiving 6 weeks of physical therapy). The following parameters were taken at baseline, 2, 4, and 6 weeks: visual analog scale (VAS) scores of present pain; lowest and highest pain over the past period; percentage of pain relief; jaw function assessment by the Mandibular Function Impairment Questionnaire (MFIQ); and pressure pain thresholds (PPTs) of the masseter, temporalis, and thumb muscles. Statistical analysis used a linear mixed model and corrected for multiple testing (Tukey test). RESULTS: Pain and MFIQ scores decreased while PPTs increased in both groups. Only after 4 and 6 weeks, significant differences were present for the PPT of the masseter in group I (P < .02) and the temporalis in both groups (P < .01). Also, the VAS scores of present (P < .02), minimal (P < .01), and maximal (P < .0001) pain and the MFIQ score (P < .001) improved. After 6 weeks, a mean of 60% pain decrease was reported (P < .0001). There were no significant differences between the groups receiving 4 weeks vs 6 weeks of physical therapy. CONCLUSION: A conservative approach involving counseling and physical therapy resulted in significant improvement in parameters of pain and jaw function in patients with myofascial pain. A controlled study will be necessary to elucidate the specific effectiveness of physical therapy over counseling or no treatment. PMID- 12756931 TI - Predictors of bruxism, other oral parafunctions, and tooth wear over a 20-year follow-up period. AB - AIMS: To analyze predictors of bruxism, other oral parafunctions, and tooth wear in a group of subjects who had been examined 20 years earlier. METHODS: Originally, 402 randomly selected 7-, 11-, and 15-year-old subjects were examined clinically and by means of a questionnaire. Twenty years after the first examination, 94% of the original group could be traced, and 320 (85%) completed and returned the questionnaire. Of the oldest group, 100 (81%) also underwent a clinical examination focusing on occlusal factors and function and dysfunction of the masticatory system. For analyses of predictors of some oral parafunctions and tooth wear registered at the 20-year follow-up, logistic regression was used with recordings at the first examination as independent variables. RESULTS: Subjective reports in childhood of bruxism (defined as tooth clenching during daytime and/or tooth grinding at night), clenching only, grinding at night only, nail biting, and/or other parafunctions were predictors of the same oral parafunctions 20 years later. There were different predictors of the 2 components of bruxism, daytime tooth clenching and tooth grinding at night. Postnormal occlusion (Angle Class II malocclusion) and tooth wear in childhood predicted increased tooth wear in adulthood. Subjects with nonworking-side interference had less anterior tooth wear than those without such interference. CONCLUSION: Oral parafunctions in childhood may be a persistent trait in many subjects. Postnormal occlusion and tooth wear in childhood predicted increased anterior tooth wear 20 years later, whereas nonworking-side interference reduced the risk for such wear in 35-year old subjects. PMID- 12756932 TI - Bruxism force detection by a piezoelectric film-based recording device in sleeping humans. AB - AIMS: To test the reliability and utility of a force-based bruxism detection system (Intra-Splint Force Detector [ISFD]) for multiple night recordings of forceful tooth-to-splint contacts in sleeping human subjects in their home environment. METHODS: Bruxism-type forces, i.e., forceful tooth-to-splint contacts, during the night were recorded with this system in 12 subjects (6 bruxers and 6 controls) for 5 nights in their home environment; a laboratory based nocturnal polysomnogram (NPSG) study was also performed on 1 of these subjects. RESULTS: All 12 subjects were able to use the device without substantial difficulty on a nightly basis. The bruxer group exhibited bruxism events of significantly longer duration than the control group (27 seconds/hour versus 7.4 seconds/hour, P < .01). A NPSG study performed on 1 subject revealed that, when the masseter muscle electromyogram (EMG) was used as a "gold standard," the ISFD had a sensitivity of 0.89. The correlation coefficient between the duration of events detected by the ISFD and the EMG was also 0.89. CONCLUSION: These results suggest that the ISFD is a system that can be used easily by the subjects and that has a reasonable reliability for bruxism detection as reflected in forceful tooth-to-splint contacts during sleep. PMID- 12756933 TI - Bruxism levels and daily behaviors: 3 weeks of measurement and correlation. AB - AIMS: To test whether 3-week duration recordings of sleep bruxism are correlated with daily behaviors. METHODS: Twelve patients with a sleep bruxism disorder were monitored to see if any daily behaviors (stress, physical activity, anger), jaw pain/headache symptoms, or sleep quality were correlated with their sleep bruxism levels. A telemetric-based system was used for monitoring bruxism levels, which were detected with an intra-appliance piezoelectric film system. Bruxism was defined as a force applied to the occlusal surface of the splint at or above a level of 10% maximum voluntary contraction. Bruxism levels were recorded at night for at least 3 weeks on the 12 subjects in this study (6 females and 6 males). Patients used standard (100 mm) visual analog scaling methods during this period to rate their daily behaviors, sleep quality, and jaw-pain/headache symptoms in a diary. Correlation analysis was performed between these recorded variables. RESULTS: The subjects demonstrated both bruxism and sleep disturbance, and the mean bruxism score for the male subjects was significantly higher than that for the female subjects. Overall, no single diary variable was consistently correlated with the bruxism levels in these subjects. CONCLUSION: These data support the conclusion that bruxism is not strongly related to any of the subject's self-monitored daytime activities or sleep quality. PMID- 12756935 TI - Tenth world congress on pain. San Diego, California, August 17-22, 2002. PMID- 12756934 TI - Chronic paroxysmal hemicrania: a case report and review of the literature. AB - Chronic paroxysmal hemicrania (CPH) is a rare type of headache that is characterized by daily, multiple, short-lasting attacks of severe pain and associated autonomic symptoms. The pain is strictly unilateral and presents most commonly in the ocular, temporal, maxillary, and frontal areas. The excruciating, throbbing pain of CPH can be misdiagnosed as pain associated with dental pathology, especially when located in the maxillary area. Moreover, pain manifesting in the maxillary and temporal areas can be confused with temporomandibular disorders. CPH patients occasionally seek treatment in dental offices or orofacial pain centers. Accordingly, dentists should be familiar with CPH in order to avoid unnecessary, irreversible dental treatment. A case is presented to highlight many of the features of CPH. PMID- 12756936 TI - Nation unprepared for microbial threats, IOM reports. PMID- 12756937 TI - Patient safety is primary job for Missouri pharmacist. PMID- 12756938 TI - Asheville Project improves patient outcomes, cuts medical costs. PMID- 12756939 TI - Webographies: a patient's guide to health information on the Internet. PMID- 12756941 TI - Resolution of drug-related problems in home care patients through a pharmacy referral service. AB - A referral-based, pharmacist-conducted medication management program designed to identify, categorize, and resolve drug-related problems (DRPs) in a home health care (HHC) population was studied. A clinical pharmacy service model was developed to identify patients at high risk for adverse health events resulting from DRPs. Policies and procedures were developed for this service model, including explicit referral criteria, patient-consent documentation, and physician orders. All identified DRPs were categorized, and pharmacist activities relating to the discovery and resolution of DRPs were tabulated. From July 1, 2001, through March 29, 2002, 80 patients were referred for and received clinical pharmacy services, and pharmacists identified 271 DRPs. Every patient referred had a DRP, and 32% of DRPs were identified by a visit to the patient's home. Nearly 65% of pharmacist recommendations were implemented. Using a referral-based practice model in an HHC setting, pharmacists were able to identify and resolve DRPs for every patient referred. PMID- 12756940 TI - Current approaches to the prevention and treatment of postmenopausal osteoporosis. AB - Current approaches to the prevention, detection, treatment, and monitoring of postmenopausal osteoporosis are discussed. In the United States, 44 million men and women ages 50 years or older have low bone mass or osteoporosis. The most devastating consequence of this disease is fractures. The assessment of osteoporosis risk includes determining risk factors, conducting laboratory and physical examinations, and measuring bone density and bone-turnover markers. Once risk has been established, nonpharmacologic strategies, such as exercise, appropriate dietary habits, and discontinuing tobacco and alcohol use, are helpful. Fall prevention and adequate intake of calcium and vitamin D are critical. When pharmacologic therapy is warranted, bisphosphonates have shown the greatest benefit in preventing bone loss and lowering fracture rates. Selective estrogen-receptor modulators and calcitonin are also options for prevention or treatment of osteoporosis. Estrogen should not be used for the sole purpose of osteoporosis prevention; however, short-term use is acceptable for women with vasomotor symptoms or in whom the benefits outweigh the risks. Parathyroid hormone may offer another treatment alternative. A variety of pharmacologic options are available for patients with osteoporosis in whom lifestyle modifications have proven insufficient. Bisphosphonates are the mainstay of drug therapy. PMID- 12756942 TI - Using personal digital assistants to document pharmacist cognitive services and estimate potential reimbursement. AB - The use of personal digital assistants (PDAs) to document pharmacist cognitive services and estimate potential reimbursement was studied. Between September 2001 and February 2002, four pharmacy residents and four clinical pharmacists used PDAs for documenting cognitive services. Interventions recorded on paper during the same six-month period one year earlier were reviewed for comparison. Potential reimbursement for these services was calculated by linking current procedural terminology codes and charges to the electronically documented services. Over the six-month study period, pharmacists recorded 7319 interventions with PDAs, compared with 5028 documented on paper during the earlier six-month period. Potential claims for pharmacists' cognitive services documented with PDAs amounted to more than $1 million in the six months, assuming a 100% reimbursement rate. PDAs provide a simple, efficient paperless system for documenting pharmacists' clinical services and generating reimbursement claims. PMID- 12756943 TI - Stability and preservative effectiveness of treprostinil sodium after dilution in common intravenous diluents. AB - The stability of treprostinil sodium after dilution in three common i.v. infusion vehicles was assessed. The chemical stability of treprostinil sodium was tested over a 48-hour period at 40 degrees C and 75% relative humidity after dilution in each of three diluents: sterile water for injection, 0.9% sodium chloride injection, and 5% dextrose injection, and after passage through an i.v. delivery system. Chemical analysis was conducted by using a validated stability-indicating high-performance liquid chromatographic assay, visually inspecting the solutions, and measuring the pH of each solution. The preservative effectiveness of the solutions was tested by the recovery of inoculations of compendial microorganisms after 48 hours in dilute solutions of treprostinil sodium. All assay results for treprostinil were within 90.0% to 110.0% of the prepared solutions diluted at 0.004 and 0.13 mg/mL treprostinil sodium in sterile water for injection and 0.9% sodium chloride injection. The assay results were the same for dilute treprostinil solutions in 5% dextrose injection at concentrations of 0.02 and 0.13 mg/mL. The pH values for these solutions remained within acceptable values of 6.0 to 7.2 for the stability study. No change in physical appearance or any visible particulate matter was observed. Approximately 70% of metacresol, the preservative, in the dilute treprostinil sodium solutions was removed before reaching the terminal end of the tubing. None of the dilute treprostinil sodium solutions supported microbial growth in the cassette reservoirs for the organisms considered. Treprostinil sodium 0.13 mg/mL solution in sterile water for injection, 0.9% sodium chloride for injection, and 5% dextrose for injection appeared to be stable after storage in controlled ambulatory drug-delivery systems for 48 hours at 40 degrees C and 75% relative humidity. Treprostinil sodium 0.004 mg/mL in sterile water and 0.9% sodium chloride for injection and 0.02 mg/mL in 5% dextrose injection was also stable under the same conditions. None of the solutions showed signs of microbial growth. PMID- 12756944 TI - Effect of changing s.c. epoetin alfa administration from thrice weekly to once weekly in hemodialysis patients. PMID- 12756945 TI - Management of community-acquired pneumonia at a tertiary care medical center in Lebanon. PMID- 12756946 TI - Standards for continuing pharmacy education: role of the American Council on Pharmaceutical Education. PMID- 12756947 TI - Maintaining quality pharmaceutical education in the digital age. PMID- 12756948 TI - Impact of guidelines on vancomycin use at a Hong Kong teaching hospital. PMID- 12756949 TI - Introducing interdisciplinary collaboration to nursing and pharmacy technician students. PMID- 12756951 TI - Pharmacy as a medical specialty. PMID- 12756950 TI - Interpretation of findings about dalteparin interchange. PMID- 12756952 TI - [Canine distemper virus--an agent looking for new hosts]. AB - Canine distemper is caused by the canine distemper virus (CDV), a RNA virus belonging to the genus Morbillivirus of the family Paramyxoviridae. The genus Morbillivirus includes measles virus, Rinderpest virus and peste-des-petits ruminants virus. The host spectrum of CDV, which includes numerous families of Carnivores, has been changed in the last years and distemper-like diseases have been observed in numerous other species. These include epidemics in large felids, marine mammals and javelinas. Different viruses have been isolated from pinnipeds including a seal-specific isolate, termed phocine distemper virus 1, PDV-1, and a CDV strain, named PDV-2. Retrospective analysis of previous epidemics among marine mammals in various regions of the world provide evidence for the occurrence of so far unrecognized morbillivirus epidemics. In some including the mass mortalities of Baikal and Caspian seals and of large felids in the Serengeti, terrestrial carnivores including dogs and wolves have been suspected as a vector for the infectious agent. However, in other epidemics among marine mammals the source of infection remains unknown including both seal epidemics in northwestern Europe in 1988 and 2002. It remains to be determined whether a morbillivirus from other marine mammals or terrestrial carnivores caused the infection in this unprotected seal populations. PMID- 12756953 TI - [Electron microscopic investigation of CD4+ lymphocyte cell line C8166 after infection with simian immunodeficiency virus (SIV)]. AB - The SIV infection of rhesus macaques (Macaca mulatta) is the most appropriate animal model in HIV research. The permanent human T-cell line C8166 is used for in vitro SIV propagation. This paper describes ultrastructural features of the cells after infection with SIVmac. The C8166 cells are ultrastructurally characterized by a heterogenous morphology which is independent of the infection. SIV induced cell syncytia are observed 18 hours after infection. Viral particles and budding occur 48 hours p.i with a peak at the day 8. Viral particles present the typical lentiviral morphology. Using the monoclonal antibody anti SIVp28 and ultra small (0.8 nm) immunogold-silver enhancement technique, we are able to demonstrate SIV antigen immunoelectron microscopically. Therefore, this ultrastructural method is suitable to detect SIV antigen in in vivo experiments with C8166 cells from day 8 p.i. serving as positive control. PMID- 12756954 TI - [Bronchopneumonia and polyarthritis due to Mycoplasma bovis in a calf]. AB - This case report describes gross lesions and histopathological findings in a 3 months-old calf originating from a feedlot with approximately 400 cattle. In this animal and additional 14 cattle of similar age, which were kept together in the same stable, swollen joints had occurred suddenly. The examination of this calf showed that a severe polyarthritis induced by haematogenous spread of Mycoplasma bovis following bronchopenumonia was present, which was characterised by necrotising lesions of the joint capsules and severe cartilage erosions. PMID- 12756955 TI - Cutaneous lesions in experimental acute and subacute African swine fever: an immunohistopathological and ultrastructural study. AB - Histopathological, immunohistochemical and ultrastructural cutaneous changes are described in acute and subacute forms of experimental African Swine Fever (ASF). Fourteen 3-month-old Large White cross pigs were infected intramuscularly with the Dominican Republic 78 (DR78) ASF virus isolate and euthanized in pairs on alternatives days (3 to 17) post-inoculation (dpi). Three pigs were found dead at 8, 10 and 13 dpi, respectively. Antibodies against viral antigen Vp73, human fibrinogen, glycoprotein IIIa and Factor VIII-ra were used to evaluate viral antigen distribution, fibrin microthrombi and platelets in dermal vessels, respectively. Cutaneous lesions were characterised by vascular changes ranging from hyperaemia, mild oedema, scarce fibrin microthrombi and microhaemorrhages in euthanized animals, to generalized fibrin microthrombosis and microhaemorrhages in dead pigs. Secondary pustules and superficial folliculitis were observed in two animals dead at 10 and 13 dpi. Diffuse cytoplasmic Vp73 labelling was found in numerous intravascular monocytes and dermal macrophages. Ultrastructural studies showed mature viral particles in the lumen of dermal vessels but viral replication was not observed; nonetheless, microtubuloreticular structures were observed in the cytoplasm of some endothelial cells and macrophages which showed cytopathic effects, signs of cell activation or degeneration. Morphological and immunohistochemical evidences of platelet activation, degranulation and consumption were observed from 5 dpi onward. PMID- 12756956 TI - [Tumors of the thyroid gland in dogs--a local characteristic in the area of Leipzig]. AB - During 17 years (1985-2002) 4.072 necropsies in dogs were performed at the Institut fur Veterinar-Pathologie of the University of Leipzig. 154 of them showed tumors of the thyroid. Retrospective classification of the neoplasia between 1992-2002 revealed 6 adenomas, 18 follicular, 33 solid-follicular and 21 solid carcinomas as well as one squamous cell carcinoma, one medullary (C-cell) carcinoma and one fibrosarcoma. During the time investigated, the number of thyroidal neoplasia out of all tumorous diseases in dogs decreased from 48% in 1986 to 3.8% in 2002. This phenomenon may be the result of an increased use of commercially produced animal food with supplementation of iodine. This may have led to the disappearance of the "typical" disease--thyroidal cancer--in dogs in this endemic area of iodine deficiency. The use of immunohistochemical markers shows that the expression pattern in neoplastic follicular areas mostly corresponds to the intact thyroidea. But in solid tumor regions the expression of thyroglobulin, cytokeratin 19 and neuron specific enolase (NSE) has diminished in most areas. Thyroid transcription factor 1 (TTF-1) is expressed in all carcinomas in variable intensity. Medullary carcinoma typically express calcitonin. PMID- 12756957 TI - [Acute herpesvirus-gastritis in a cat]. AB - Gastritis in cats is caused, among other things, by infectious agents, like bacteria, metazoic parasites or viruses. Herpesvirus-gastritis has not as yet been documented in cats. Therefore in this paper such a case will be described. In this case the mucous membrane of the stomach shows multifocal acute necroses with evidence of intranuclear inclusion bodies in epithelial cells of the gastric glands. By means of electron microscopy the causative virus can be specified as herpesvirus. PMID- 12756958 TI - [Experimental studies on the therapy of epiglottis hypoplasia in horses- transendoscopic injection of collagen and polytetrafluoroethylene]. AB - Epiglottic augmentation by transendoscopic injection of an implant material was performed on ten clinically healthy horses. In six cases bovine collagen (Zyplast) was used, in the remaining four horses the injection was done with polytetrafluorethylene (PTFE-Paste). The results of the surgery were observed endoscopically and by contrast radiography. Using the radiographs, epiglottic length and thickness were measured. For necropsy and histologic assessment the horses, three and two animals of the two groups, were euthanized at three and 12 weeks after surgery. The data obtained from the measurements at necropsy were correlated with those from the radiographs. The minimal invasive technique was very easy to perform with the bovine collagen while PTFE was less suitable for the transendoscopic injection. The data concerning the epiglottic length as well as its thickness measured on the radiographs correlated well to those obtained from the measurements at necropsy. After injection of collagen as well as PTFE an organisation of the implant was noticed histologically, which increased in accordance with the increasing time after injection. As far as the collagen implant is concerned, there was fibrous granulation tissue accompanied by an inflammatory reaction consisting mainly of lymphocytes and histiocytes. In contrast the injection of PTFE led to the development of a foreign body granuloma. In addition, some cases showed ulcers. Both implant materials led to an increase in epiglottic thickness, so that one can expect an improvement in its stability which could be of benefit to the therapy of epiglottic hypoplasia. But this increase in thickness had its maximum level directly after injection and declined over the period observed. Still, especially in the case of the collagen used here, the development of a fibrous granulation tissue and the immigration of fibroblasts and fibrocytes into the implant indicates that there is a stabilisation to some extent. Bovine collagen was proven to be highly suitable for transendoscopic injection. Although there is report about good clinical results using PTFE via laryngotomy, this material seems to be unsuitable for the transendoscopic technique due to the circumstances that lead to a higher number of complications. PMID- 12756959 TI - African swine fever and classical swine fever: a review of the pathogenesis. AB - This paper describes major pathogenetic mechanisms of African and Classical Swine Fever virus infections. The interactions between both viruses and the monocyte macrophage-system result in the release of mediator molecules, which are important for the further progression of the diseases. The causes of the thrombocytopenia and the mechanisms of the haemorrhages, which are characteristic in both infections, are described. Apoptotic cell death is regarded as the predominant cause of lymphopenia in both virus infections. PMID- 12756960 TI - [Effects of excess caloric fat feeding on the lipid metabolism in Shetland ponies]. AB - To investigate the influence of overweight and dietary fat supplementation on lipid and insulin glucose metabolism of Shetland ponies, eight Shetland pony geldings were fed a hypercaloric (30 MJ DE/150 kg bwt. and day) fat diet (10% fat as soybean oil) or a carbohydrate control diet for nine months until ponies gained an overweight of 15%. Afterwards oral glucose tolerance tests (oGTT; 5, 6 mmol/kg bwt.) were performed after a 12 hour fast and after a fast which led to an increase of plasma triglyceride concentrations to a threshold of 3 mmol/l (36 65 hrs.). Plasma concentrations of glucose, insulin, triglycerides and non esterified fatty acids (NEFA) were determined for 480 minutes after the glucose load. Ponys having had received the control diet tended to a higher insulin secretion in case of both oGTTs, whereas the glucose tolerance was similar in both groups but lower than in ponies of normal weight. During the oGTTs after fasting leading to the plasma triglyceride threshold, triglyceride concentrations decreased significantly (p < 0.05) faster and stronger in fat fed ponies. Additionally, fat fed pony showed significantly (p < 0.05) lower NEFA levels. The results of this study demonstrate a positive effect of fat feeding on the triglyceride clearance of overweight Shetland ponies. PMID- 12756961 TI - Adrenocortical carcinoma in a donkey. AB - An adrenocortical carcinoma was found in an eight-year-old male donkey. The main clinical feature was severe depression and inappetence. The animal died 24 hours after hospitalization. Necropsy revealed a large tumor around the anterior pole of the left kidney and metastases to liver, heart and lung. Fibrous adhesions were seen between the tumor and the intestinal serosa. The abdominal cavity contained a large amount of red-brown fluid. PMID- 12756962 TI - Pleural mesothelioma in a young cat. AB - A mesothelioma was seen as an incidental finding in the thoracic cavity of an eleven-month old female cat. At necropsy, the pale nodular lesions were spread over the ventral parts of the lungs and their corresponding parts of the diaphragm and the thoracic wall. Microscopically, the tumor consisted of mesenchymal tissue with large amounts of collagenous fibers covered by a mainly unilayered, polymorphic, partly vacuolated line of cells with large nuclei. Mitotic figures were rare. Based on morphological appearance the lesion was classified as an early mesothelioma. PMID- 12756963 TI - [Treatment of emergencies in the hospital--problems and management]. AB - Due to the growing number of high-risk patients, the increasing proportion of geriatric patients and the expansion of surgical and invasive-diagnostic procedures, medical stuff in hospitals are confronted with a rising number of emergency situations. Nearly 50% are of cardio-circulatory origin and occur during surgical interventions or immediately afterwards. Another cause of life threatening complications are side-effects of orally or intravenously administered agents, especially after treatment with antibiotics, anaesthetics, analgetics and sedatives. Due to a lack of emergency training and management in most hospitals, the survival rate after cardiopulmonary resuscitation in general wards lies between just two and 35%. Thus it seems necessary to perform special training in CPR procedures and emergency management at regular intervals for the entire medical stuff. In addition, a special infrastructure for giving sufficient treatment in emergencies has to be established (emergency team, emergency telephone number, intra-hospital emergency car). The second part of this review presents current diagnostic and therapeutic strategies for the most common emergency situations, e.g. anaphylaxis, myocardial infarction, pulmonary embolism, gastrointestinal bleeding, and heparin-induced thrombocytopenia (HIT). PMID- 12756964 TI - [Clinical results with the "open lung concept"]. AB - Elements of the "open lung concept" are being increasingly included in clinical ventilatory strategies. Despite encouraging experimental investigations to date, relatively few studies exist that examine the clinical application of the complete concept. The aim of this study was to prove that with effective recruitment maneuvers and titrated PEEP levels this concept is applicable in clinical settings. We sought to determine if it was possible to achieve a significant improvement in oxygenation and also to examine what side-effects resulted. Twenty consecutive patients who had had an acute lung injury (ALI) for less than 72 hours, with an oxygenation index (P/F-Ratio = quotient from arterial partial pressure of oxygen [PaO2] and the inspiratory fraction of oxygen [FiO2]) of less than 200 torr, and with a PEEP > or = 10 cmH2O were treated using a recruitment manoeuvre (RM). A PEEP was titrated to keep the lung open, and the patients were kept under pressure-controlled ventilation. The P/F-Ratio increased while using a recruitment pressure of 66 +/- 13 cmH2O from 137 +/- 41 to 381 +/- 150 torr (p < 0.001). The titrated PEEP which kept the lung open after recruitment was 17 +/- 3 cmH2O. One patient developed a pneumothorax. The dose of norepinephrine was increased in ten patients from 0.24 +/- 0.12 to 0.31 +/- 0.1 microgram/kg/min. Due to elevated liver enzymes within the first 48 hours, titrated PEEP had to be decreased in three patients. The clinical application of the "open lung concept" demonstrated a quick and effective improvement in oxygenation in many patients. Side-effects in some patients limited the use of high PEEP levels. PMID- 12756966 TI - [Atraumatic retroperitoneal hemorrhage--interdisciplinary and differential diagnostic considerations based on a case report]. AB - The differential-diagnostic strategy and the order of precedence of most diverse radiologic diagnostic procedures are discussed based on a case of retroperitoneal bleeding. Apart from iatrogen-caused bleedings, the therapeutic anticoagulant therapy in the context of the patient's disease, haemodialysis or a rare, spontaneously-occurring retroperitoneal bleeding play a substantial role. In the order of precedence of radiologic diagnostic procedures for fast diagnosis of a retroperitoneal haematoma, the abdomen CT-scan is the preferred method. PMID- 12756965 TI - [Is the combination of remifentanil and propopfol suitable for transsphenoid resection of the hypophysis?]. AB - In a multi-center trial, the feasibility of combining remifentanil (RF) and target-controlled infusion of propofol (P) for patients undergoing transsphenoidal resection of the pituitary gland was tested. After IRB approval, 74 patients (29 male/45 female) were included in the study. The concentration of RF and the target concentration of P were recorded as were heart rate (HR) and mean arterial blood pressure (MAP). For intubation the RF dosage was 0.26 +/- 0.06 microgram.kg-1.min-1 and the target concentration of P was 3.16 +/- 0.63 micrograms.ml-1. After induction, HR and MAP decreased significantly. The painful events of the operation were preparation of the nasal mucous membrane and penetration of the sella turcica. By adjusting the RF dose to 0.31 +/- 0.09 microgram.kg-1. min-1 and the target concentration of P to 3.48 +/- 1.49 micrograms.ml-1, an increase of HR and MAP above initial values was avoided at this time. Hypotension and bardycardia were treated in eight patients (10.8%) with a vasopressor, in four patients (5.4%) with atropine and in four more patients (5.4%) with a combination of these drugs. Two patients (2.7%) needed antihypertensive therapy. The average time interval between the end of P-TCI and spontaneous breathing was 6 +/- 3 min (median 6 min) and till patients opened their eyes 9 +/- 4 min (median 9 min). After 13 +/- 4 min (median 13 min) the patients became orientated. The average doses of analgetics were 19.5 +/- 19.9 mg piritramide and 1.8 +/- 1.0 g metamizol during the first 12 hours postoperatively. Eight patients (10.8%) did not need any analgetics. We suggest that the combination of RF and P as a "fast track concept" can supplement the repertoire of anaesthetic managements used for transsphenoidal resection of the pituitary gland. PMID- 12756967 TI - How does quality enter into health care purchasing decisions? PMID- 12756968 TI - Validating polymerase chain reaction for detecting HPV in cervical intraepithelial neoplasia. AB - OBJECTIVE: To validate polymerase chain reaction (PCR) analysis for detecting HPV in Mexican women with cervical intraepithelial neoplasia, grade 2 and 3 (CIN 2/3) versus histologic evidence. STUDY DESIGN: A diagnostic test study was carried out. A sample of 25 selected women who were diagnosed by histology as having CIN 2/3 was analyzed. Biopsies were examined for HPV infection using light microscopy. The histologic criteria used for HPV infection included koilocytosis, dyskeratosis cells, bi/multinucleation, and parakeratosis. PCR was performed on each sample using commercial probes (MY09 and MY11), and then HPV typing was carried out by restriction fragment length polymorphism analyses. RESULTS: PCR revealed that 88% (22/25) of the women were HPV positive (19 high risk and 3 low risk). In contrast, histology revealed that 28% (7/25) of the women were HPV positive. The number of women infected with HPV was 3.14 times (88/28) more frequently detected with PCR procedure than with the histology. Using PCR as the gold standard, 4 values (true positive, false positive, false negative and true negative) were obtained (7, 0, 15 and 3), and histology had a sensitivity, specificity, and positive and negative predictive values of .32, 1.00, 1.00 and .17, respectively. There was a correlation between low-risk and high-risk for PCR (chi 2 with Yates correction = 6.32, P = .012). CONCLUSION: PCR is a powerful tool for the early detection of HPV infection and is independent of histologic criteria. PMID- 12756969 TI - Molecular weight, tertiary structure, water binding and colon behaviour of ispaghula husk fibre. AB - Molecular variables, using aqueous and alkaline extracts, of the polysaccharide from ispaghula husk (IH) were examined using gel-permeation chromatography linked to multi-angle laser light scattering. Progressive extraction can yield a component with a molecular weight (MW)value up to about 7 x 106 Da, and gels, which accompany the extraction, have MW ranging from 10-20 x 106 Da. To mimic the polysaccharide degradation, particularly in the colon, the solid IH was degraded progressively using ionising radiation. A chain break occurs every 7.5 kGy in NaOH and every 15 kGy in water. The solid-state matrix is opened by the radiation to yield increased visco-elasticity of the aqueous extracts at critical radiation doses, before further degradation occurs after about 12 kGy. Differential scanning calorimetry is used to study the mechanism of interaction of water with IH. The first water to be taken up is non-freezing water and represents about twelve water molecules/disaccharide unit of the polysaccharide. As the water content is increased, the water becomes bound to the polysaccharide and freezes and melts at a temperature different from free water. This water is thermodynamically distinguishable from free water. It forms amorphous ice on cooling which crystallises exothermically and subsequently melts endothermically. Saturation occurs at a water content of 2-3 g water/g polymer, showing that about 60% of the water in the system is 'bound'. The most surprising conclusion is that despite the fact that the IH swells in water to form a solid and stiff gel, the greater part of that water in the gel is still free and behaves like liquid water. PMID- 12756970 TI - Primary structure of arabinoxylans of ispaghula husk and wheat bran. AB - The primary structures of ispaghula husk and wheat bran were investigated in order to determine how and why these fibres are among the most beneficial dietary fibres. To this end, the polysaccharide preparations have been subjected to enzymic hydrolysis and methylation analysis.The results have shown ispaghula husk and wheat bran to be very-highly-branched arabinoxylans consisting of linear f-D (1-4)-linked xylopyranose (Xylp) backbones to which a-L-arabinofuranose (AraJ3 units are attached as side residues via a-(l13) and a-(1-02) linkages.Other substituents identified as present in wheat bran include P-D-glucuronic acid attached via the C(O)-2 position, and arabinose oligomers, consisting of two or more arabinofuranosyl residues linked via 1-2, 1-3, and 1-4 linkages. Ispaghula husk arabinoxylan is more complex having additional side residues which include a D-glucuronopyranose (GalAp)-(1-42)-linked-a-L-rhamnopyranose-(1-04)-0-D-Xylp, a-D GalAp-(l-o3)-linked-a-L-Araf-(l-4)-[3-D-Xylp, and a-L-Araf-(l-43)-linked-P-D-Xylp (1l -4)--D-Xylp. The beneficial effects of increased faecal bulk and water holding capacity are undoubtedly related to the structures of the arabinoxylans, with differences in their efficacy to treat various functional bowel disorders due to their specific structural features. PMID- 12756971 TI - Fibre and water binding. AB - The range of interactions between fibre and water and the consequential properties of the bound water are modelled and examined. Dietary fibre may interact with water by means of polar and hydrophobic interactions, hydrogen bonding and enclosure. The results of these interactions vary with the flexibility of the fibre surface. When the fibre is insoluble or junction zones are formed,this may result in profound changes in the surrounding water. Such interactions are capable of affecting the structuring and solvation properties of water well away from the immediate surfaces involved. In particular, the specific properties of water enclosed by dietary fibre are examined, an area of investigation previously receiving scant attention. The way this enclosure may affect the properties of water is exemplified by modelling the colon to show how fibre may exert a beneficial action by the preferential partitioning of hydrophobic carcinogens. Unfermented dietary fibre has a tendency to form low density expanded water that acts as a preferential solvent for hydrophobic molecules when compared with the less-structured denser water within the much more hydrophilic mucus layer. PMID- 12756972 TI - Abnormalities of the cadherin-catenin complex in chemically-induced colo-rectal carcinogenesis. AB - Beta-Catenin is a multifunctional protein originally identified as a component of the cadherin cell-cell adhesion complex. It also binds the adenomatous polyposis coli (APC) tumour suppressor which controls beta-catenin cellular levels through its degradation. (beta-Catenin and/or APC mutations result in increased cytoplasmic Beta-catenin and nuclear translocation. The aim of the present study was to examine the expression and cellular localisation of alpha and beta catenin, p120 and E-cadherin in a chemically-induced mouse model of colo-rectal cancer using 1,2-dimethylhydrazine (DMH). Female Balb/C mice were injected subcutaneously with a solution providing 25 mg DMH base/kg body weight for 17 weeks. Animals were killed and tumours identified in the intestine with a dissecting microscope. Formalin-fixed paraffin-embedded sections of normal and dysplastic colonic mucosa were stained by an indirect avidin-biotin immunohistochemical technique using mouse monoclonal antibodies, and membranous, cytoplasmic and nuclear cellular localisation was assessed by light microscopy. Staining distribution scored as follows: 3, > 90 % positive epithelial cells; 2, >50 % positive epithelial cells; 1, <50 % positive epithelial cells. Non dysplastic colonic epithelial cells revealed beta-catenin expression at the membrane (33/41 scored 3),areas of cytoplasmic expression (24/41 scored 1) and no nuclear staining. Dysplastic colonic epithelium revealed increased membranous and cytoplasmic, beta-catenin immunoreactivity (39/41 and 38/41 both scored 3) with focal nuclear staining (14/41). Expression patterns for ac-catenin, p120, and E cadherin were similar to beta-catenin with increased membranous and cytoplasmic immunoreactivity in dysplastic mucosa, although no nuclear staining was observed. Increased cytoplasmic expression and nuclear localisation of beta-catenin are consistent with a possible mutation in its gene, and this finding was in keeping with the mutational analysis of exon 3 by single-strand conformational polymorphism. Increased immunoreactivity of the other catenins also suggests further disruption in catenin regulation. In summary, alterations in the beta catenin expression and cellular localisation in the DMH-induced tumours are similar to those seen inhuman sporadic colorectal tumours. The DMH is therefore a useful model for studying the abnormalities of the E-cadherin-catenin pathway in colorectal carcinogenesis. PMID- 12756973 TI - Colonic mucin: methods of measuring mucus thickness. AB - Mucus is a water-insoluble gel secreted by the gastrointestinal tract. It exists as a protective gel layer adherent to the epithelial surface of the stomach, small intestine and colon. The mucus gel is composed of 1-10 % (w/v) mucin glycoprotein in a plasma-like fluid. Since the mucus gel is predominantly water, standard histological techniques dehydrate the mucus, making visualisation of the functional barrier difficult. Specialist techniques have been developed to enable visualisation of the intact mucus layer. A simple histological method using snap frozen tissue, sectioned with a cryostat and stained with modified periodic acid Schiffs/Alcian blue in mucus-preserving conditions will be described. A second powerful in vivo animal model is described which enables measurement of mucus secretion over time. The use of these two methods has allowed the characterisation of the normal mucus layer in the colon and the determination of how it is affected by disease and dietary intervention, in particular the effect of dietary fibre, and evidence that fibre deficiency results in colonic mucosal fragility is presented. PMID- 12756974 TI - Colonic mucus: secretion and turnover in relation to dietary fibre intake. AB - The colon is subjected to a myriad of potentially damaging agents that may reside within the lumen for 1-2 d. Its first line of defence against these agents is the protective mucus bilayer that lines the entire colonic mucosa. This bilayer acts as a physical barrier to mucosal aggressors and also reduces shear stress to the mucosa. These actions are dependent on the unstimulated ('resting') colonic mucus thickness, and also on the rate that this layer can be replenished. The colonic mucus layer is altered in a number of colonic diseases that have been linked to a deficiency of fibre in the diet. The action of fibre intake on colonic mucus thickness and secretion is unknown. Using an in vivo rat model it has been demonstrated that: (1). fibre deficiency leads to a decreased protective potential of the mucus layer (e.g. the mean resting mucus thickness of the fibre deficient group (429 microm) was significantly lower than its respective control (579 microm; P< 0-001), as was its total mucus secretion over 6 h (270 microm v. 541 microm; P<0-01); (2). specific fibre types in the diet alter the secretion dynamics of colonic mucus (e.g. a cellulose-based diet reduces total mucus secretion over 6 h compared with its control (175 microm v. 463 microm). Analysis of the diets suggested a necessity for both soluble and insoluble fibre types in the diet to increase mucosal protection. PMID- 12756975 TI - [Two editors in dialog. The example of Justus von Liebig (1803-1873) and Johann Bartholomaus Trommsdorff (1770-1837) and their letters]. AB - Today, scientists willing to publish results face a wide range of requirements (instructions for authors, peer review procedure). During the 18th and 19th century manuscripts were treated in a less formal way. However, the scientific discussion was vital and implemented intensely via various means, especially by letters. This study analyses the dialog of two distinguished contributors to chemical and pharmaceutical sciences, both experienced journal editors, from January 1828 to February 1837. PMID- 12756976 TI - [Counseling is a multifaceted and widespread helping process]. PMID- 12756977 TI - Reproductive intentions of the newlywed Bulgarian families--artificial neural network approach. AB - INTRODUCTION: The aim of this study was to describe the reproductive intentions of newlywed Bulgarian families, build predictive models of the desired number of children by artificial neural networks (ANN) and compare these models' overall prediction accuracy with conventional statistical analyses. MATERIAL AND METHODS: Data from a large cross-sectional study on family planning knowledge and habits of newlywed couples from the region of Plovdiv (Bulgaria) in the year 2000 were used. A subset of 17 variables was extracted from the database, in order to construct and train a back-propagation multilayer ANN. Desired number of children, separately for male and female spouses, was used as an output variable. The following statistical analyses were used: descriptive statistics, percentage distribution, nonparametric tests and logit regression analysis. RESULTS AND DISCUSSION: Factors that determine the opinion of male and female spouses on desired number of children have different relative importance in the phase space. The reproductive planning of the newly-married family can be better modeled and predicted in women than in men. The reproductive intentions of the modern newlywed Bulgarian family are decided on the basis of mutual agreement and understanding. There is no difference between desired number of children by both partners. The age at marriage of spouses is increasing with a tendency to reach the Western European patterns. Artificial neural networks are found to be a very useful tool in cross-sectional population studies for factor classification and outcome modeling, especially in cases where data are not entirely suitable for logistic regression analyses. PMID- 12756978 TI - Development of pro- and antisaccades in children with attention-deficit hyperactivity disorder (ADHD) and healthy controls. AB - To date, the investigation of the antisaccade task, a simple test of "executive functions," in children with ADHD has yielded inconsistent results. The present study aimed at contributing to this issue by (a) the investigation of a large sample of carefully diagnosed ADHD patients aged 7-15 years, and (b) the analyses of differential age effects in patients and controls. Healthy control children were pairwise matched with patients (N = 46; age = 136 +/- 24 months) for age and gender, and did not significantly differ in IQ. Horizontal pro- and antisaccades were elicited under the 200-ms gap and overlap conditions (blocks of 100 trials each). Overall, patients exhibited (a) augmented pro- and antisaccadic reaction times, (b) augmented error rates (antitasks), (c) augmented proportions of early responses (all conditions), and (d) reduced proportions of express saccades under the prosaccadic gap condition. The greater decline in anti- as compared to pro SRT with increasing age that characterized controls was missing in patients. Confirming Barkley's (1997) neuropsychological theory of ADHD, these results altogether point to alterations in "executive functions" in ADHD patients that are presumably supported by frontal lobe structures, in particular the lateral prefrontal cortex and the frontal eye fields. PMID- 12756979 TI - Use of microencapsulated iron(II) fumarate sprinkles to prevent recurrence of anaemia in infants and young children at high risk. AB - OBJECTIVE: To compare the effectiveness of microencapsulated iron(II) fumarate sprinkles (with and without vitamin A), iron(II) sulfate drops, and placebo sprinkles in preventing recurrence of anaemia and to determine the long-term haematological outcomes in children at high risk of recurrence of anaemia 12 months after the end of supplementation. METHODS: A prospective, randomized, placebo-controlled design was used to study 437 Ghanaian children aged 8-20 months who were not anaemic (haemoglobin > or = 100 g/l). Four groups were given microencapsulated iron(II) fumarate sprinkles, microencapsulated iron(II) fumarate sprinkles with vitamin A, iron(II) sulfate drops or placebo sprinkles daily for six months. Primary outcome measures were change in haemoglobin and anaemic status at baseline and study end. Non-anaemic children at the end of the supplementation period were reassessed 12 months after supplementation ended. FINDINGS: Overall, 324 children completed the supplementation period. Among the four groups, no significant changes were seen in mean haemoglobin, ferritin or serum retinol values from baseline to the end of the supplementation period. During the trial, 82.4% (267/324) of children maintained their non-anaemic status. Sprinkles were well accepted without complications. At 12 months post supplementation, 77.1% (162/210) of children with no intervention remained non anaemic. This proportion was similar for children among the four groups. CONCLUSION: In most children previously treated for anaemia, further supplementation was not needed to maintain their non-anaemic status. These results may have important implications for community intervention programmes in which initial high-dose treatment is needed because of a high prevalence of anaemia. PMID- 12756980 TI - An analytical framework for the study of child survival in developing countries. 1984. PMID- 12756981 TI - Reoccurrence of levofloxacin-induced tendinitis by phenoxymethylpenicillin therapy after 6 months: a rare complication of fluoroquinolone therapy? PMID- 12756982 TI - California's new Assembly and Senate districts: geographic disparities in health insurance coverage. PMID- 12756983 TI - Dealing with drugs. PMID- 12756984 TI - [Sports and nutrition: high performance athletes need balanced energy metabolism]. PMID- 12756985 TI - [Backache in high performance sports exemplified by basketball]. PMID- 12756986 TI - [Foll, M. Reduction of excessive reactions after tuberculin skin test (Mendel Mantoux method). Gesundheitswesen 2002; 64: 544-547]. PMID- 12756987 TI - Congenital optic pit with serous maculopathy in childhood. PMID- 12756988 TI - [Leading health insurance associations and MDS present 1st German country-wide prevention report]. PMID- 12756989 TI - [Social medicine: social insurance adds committee]. PMID- 12756990 TI - [Report of the Society of "Nordwestdeutscher Chirurgen"]. PMID- 12756991 TI - [Surgery of the upper gastrointestinal tract (part 2)]. PMID- 12756992 TI - Obituary: Roy Sydney Porter (1946-2002). PMID- 12756993 TI - [Consensus statement on antimicrobial treatment of pharyngo-tonsillitis]. PMID- 12756994 TI - Surgery in Hitler's bunker. PMID- 12756995 TI - Respiration apparatus of Lavoisier. PMID- 12756996 TI - Studies of narcosis: Charles Ernest Overton introduction. PMID- 12756997 TI - [Dingeman van der Vliet, veterinary artist, 1792-1866]. AB - Dingeman van der Vliet (1792-1866) from Zierikzee in province of Zeeland, was one of the first officially trained veterinarians in the Netherlands. He was sent to Paris on the order of Louis Napoleon, king of Holland, at the expense of the Dutch government. In the period of his stay at the Veterinary School in Alfort (1808-1811) he wrote a great number of letters to his parents and friends. His letters give insight in the daily life of the school in Alfort and the festivities in Paris during the reign of the emperor Napoleon Bonaparte. In a second article, the laborious built-up of a veterinary practice in those days will be described. PMID- 12756998 TI - [Collegiality, a great blessing, but also a subject of constant concern]. AB - The incorporation of the Veterinary College into the University of Utrecht, in 1925, had large implications for the social life of the students. Their student's union, named "Absyrtus", with its own club building and longtime traditions was dissolved. The students were supposed to become members of the several unions that existed in the university. But only about a quarter of them managed to do so, and these were divided over four or five different unions. After some years voices were heard within the Netherlands Veterinary Association that collegiality, highly valued in professional life, could be hampered if no social sense was cultivated during student's days. At the same time the number of students showed a jumpy increase, so that unemployment was to be feared. The plan for a new society of veterinary students, brought forward by a commission of three professors, set up by the Veterinary Association, was accepted, and in 1932 the new society was established. PMID- 12756999 TI - [Local and regional veterinary history]. AB - Continuing a review, written in 1992, an overview is given of the five recent studies on local themes. The author emphasizes that telling a story of local traditions and usages, of striking personalities or personal experiences can illuminate the problems of a broader national or even international context. He points to the fact that the local archives contain many materials of veterinary historical relevance waiting for exploitation. PMID- 12757000 TI - Human genetics in the Holy Qur'an and Sunna. PMID- 12757001 TI - Stature in Scotland over the centuries. PMID- 12757002 TI - The decline of German medicine, 1933-45. PMID- 12757003 TI - John Gregory and the background to medical philosophy. PMID- 12757004 TI - Abstracts of the 50th Annual Meeting of the Society of Nuclear Medicine. New Orleans, Louisiana, USA. June 21-25, 2003. PMID- 12757005 TI - Abstracts of the 7th Congress of Infectious Diseases and Tropical Medicine and the 11th Annual Meeting of the German Society of Pediatric Infectious Diseases. Berlin, Germany, 27 February-1 March 2003. PMID- 12757006 TI - American Transplant Congress. Washington, DC, USA. May 30-June 4, 2003. Abstracts. PMID- 12757007 TI - Abstracts of the 6th Annual Meeting of the American Society of Gene Therapy. June 4-8, 2003. Washington, DC, USA. PMID- 12757008 TI - Abstracts of the 12th Commonwealth International Sport Conference. Manchester, United Kingdom, 19-23 July 2002. PMID- 12757010 TI - British Maternal and Fetal Medicine Society 8th Annual Conference. 20-21 March 2003. Abstracts. PMID- 12757009 TI - Abstracts of the 3rd North American Symposium on Skeletal Complications of Malignancy. April 2002, Bethesda, Maryland, USA. PMID- 12757011 TI - [Violence and the interaction between genes and environment]. PMID- 12757012 TI - Updates in Cartilage Repair. Proceedings of the 4th Symposium of the International Cartilage Repair Society. Toronto, Canada, June 2002. PMID- 12757013 TI - Proceedings of the 3rd International Conference on the Lacrimal Gland, Tear Film, and Dry Eye Syndromes: Basic Science and Clinical Relevance. November 15-18, 2000, Maui, Hawaii. PMID- 12757014 TI - [Septic encephalopathy--frequently mistaken and underestimated]. PMID- 12757015 TI - Proceedings of the 3rd S. Takahashi Memorial International Workshop on 3 Dimensional Conformal Radiotherapy. PMID- 12757016 TI - [Dizzy heights. How the heart and lungs react to hypoxic states high up in the mountains]. PMID- 12757017 TI - Toyota Motor Manufacturing, Kentucky, Inc. v. Williams: Disabling the Americans with Disabilities Act. PMID- 12757018 TI - Retinoids as regulators of gene transcription. AB - Our understanding on the molecular mechanisms that underlie complex biology of retinoids' action has increased tremendously in recent years. Retinoids can bind and activate heterodimeric receptors and can directly affect transcriptional regulation of certain genes. Alternatively, they can indirectly affect the transcriptional regulation by inhibiting certain transcription factors, such as AP-1 and STAT-3. Alterations in retinoid receptor and subsequent signaling during acute promyelocytic leukemia [APL, characterized by a specific chromosome translocation, t(15;17)], has provided invaluable tool for understanding the myeloid differentiation at molecular level. Studies with APL have highlighted the critical role that retinoids play in cancer and its treatment with retinoids. PMID- 12757019 TI - Receptor-independent induction of apoptosis by synthetic retinoids. AB - Retinoids modulate cell proliferation, differentiation and apoptosis. Many of these effects are mediated by nuclear retinoid receptors. However, studies with certain synthetic retinoids, including some that can activate retinoid receptors, revealed that they affect cell growth and especially apoptosis by mechanisms that are independent of nuclear receptors. This chapter describes the pro-apoptotic effects of the synthetic retinoid CD437 [6-[3-(1-adamantyl)-4-hydroxyphenyl]-2 naphthalene carboxylic acid], and structurally-related retinoids and summarizes the mechanisms by which they induce apoptosis. PMID- 12757020 TI - The retinoid X receptor and its ligands: versatile regulators of metabolic function, cell differentiation and cell death. AB - Retinoid X Receptors (RXRs) consist of a family of nuclear receptors that target and regulate multiple signalling pathways. The early evolutionary emergence of RXRs in comparison to other nuclear receptors may have allowed for the development of unique properties as transcriptional regulators. Indeed, the complexity of these receptors is derived from their ability to activate transcription as homodimers or as obligate heterodimeric partners of a multitude of other nuclear receptors. In addition, RXRs can regulate gene expression in a ligand-dependent (forming permissive heterodimeric complexes) or - independent (forming non-permissive heterodimeric complexes) manner. Given that ligand binding is a critical component of RXR function, this review will focus on the ligand dependent functions of RXR. The remarkably conserved ligand binding domain of RXR is a multi-functional structure that in addition to ligand binding, serves as a homo- and heterodimeric interface, and a region to bind coactivactor and corepressor molecules. RXRs have a small ligand binding pocket and therefore bind their ligands (such as 9-cis RA) with both high affinity and specificity. In the presence of ligand, permissive RXR heterodimers bind coactivators, but nonpermissive complexes can bind coactivators or corepressors depending on the activation of the RXR's heterodimeric partner. Physiologically, the temporal and tissue specific pattern of RXRs as well as the presence of phenotypic abnormalities in receptor knockout studies (most severe in RXRa -/- animals) demonstrate the important role for these receptors both during development (morphogenesis) and in adult differentiated tissues (cell proliferation, cell differentiation, cell death). These receptors also play an important regulatory role metabolic signaling pathways (glucose, fatty acid and cholesterol metabolism), including metabolic disorders such as type 2 diabetes, hyperlipidemia and atherosclerosis. RXRs function as master regulators producing diverse physiological effects through the activation of multiple nuclear receptor complexes. RXRs represent important targets for pharmacologic interventions and therapeutic applications. PMID- 12757021 TI - Retinoids in myelopoiesis. AB - The vitamin A derivative retinoic acid plays a critical role during the differentiation of myeloid progenitors towards the neutrophil lineage. This role is primarily mediated by binding of retinoic acid to retinoic acid receptor alpha (RARalpha), a nuclear receptor that modulates the expression of multiple downstream targets via retinoic acid response elements. The importance of this signalling pathway in myelopoiesis is evidenced by the recurrent disruption of the RARalpha gene by chromosomal rearrangements in all cases of acute promyelocytic leukemia (APL). Biochemical evidence suggests RARalpha performs two opposing functions, one as a repressor of gene expression in the absence of ligand, the second as a transcriptional activator in the presence of ligand, each controlled by multimeric complexes of transcription corepressors and coactivators, respectively. Here the molecular mechanisms activated by retinoic acid during myelopoiesis in the context of neutrophil development will be reviewed, together with some of the more recently identified targets of the retinoic signalling pathway. PMID- 12757022 TI - Retinoids as chemopreventive agents. AB - Retinoids are promising agents for cancer chemoprevention. The myriad effects of retinoids on biological processes including development, differentiation, homeostasis, carcinogenesis and apoptosis are mediated through their molecular targets, the retinoid and rexinoid receptors. Tissue specific expression patterns, ligand specificities, receptor numbers, their distinct functions and functional redundancy make retinoid signaling highly complex. The cross-talks of these receptors with cell surface receptors signaling pathways, as well as their interactions with multiple co-activators and co-repressors further add to the complexity of the pleiotropic effects of retinoids. Elucidation of retinoid signaling pathways and indepth understanding of the mechanisms that underlie the anti-proliferative and apoptotic action of retinoids has paved the way for designing synthetic retinoids for effective chemoprevention and therapy of cancer. Development of receptor selective synthetic retinoids is a major focus of molecular retinoid development. Other new avenues encompass identification of novel retinoid regulated genes, orphan-receptor ligands/functions, novel retinoid mechanisms involving receptor-independent apoptosis inducing activity and synergistic combinations with other agents for cancer prevention and therapy. This review focuses on recent advances in the understanding of molecular mechanisms underlying the action of retinoids and retinoid molecular targeting studies designed primarily to develop retinoids with reduced toxicity, while maintaining or enhancing activity in context of chemoprevention. The clinical efficacy of retinoid based chemoprevention trials is discussed. PMID- 12757023 TI - Retinoids in chemoprevention of cancer. AB - Retinoic acid (RA), a natural metabolite of circulating Vitamin A (retinol) and an irreversible oxidation product of retinol, is essential in maintaining the normal pathway of differentiation of epithelial tissues. RA and a number of its analogs, both natural and synthetic (retinoids), have been shown to be effective in the prevention of a variety of cancers in experimental animals, and in reversing preneoplastic lesions in humans. The retinoids exhibit a high degree of specificity in cancer chemoprevention. Diverse effects of retinoids are mediated by retinoid nuclear receptors, the ligand-inducible trans-acting transcription factors. The receptor-selective retinoids may be more effective and less toxic in cancer prevention. Our chemoprevention study with retinoids using the mouse skin carcinogenesis model indicated that retinoids are anti-tumor promoters. One of the mechanisms by which retinoids inhibit promotion of mouse skin tumor formation involves their property to inhibit the induction of ornithine decarboxylase by the phorbol ester tumor promoter 12-O-tetradecanoylphorbol-13-acetate. RARalpha and RARgamma, but not RXRs, may mediate mouse skin anti-tumor promotion activity of retinoids. Retinoids are highly selective chemopreventive agents and are toxic at high pharmacological doses. Clinical trials with retinoids should be conducted with a carefully evaluated, appropriate patient population and perhaps at low doses in combination with other chemopreventive agents with mechanisms of action different from retinoids. PMID- 12757024 TI - Clinical applications of retinoids in cancer medicine. AB - The retinoids are compounds structurally related to vitamin A. The most extensively studied agents in cancer medicine include all-trans-retinoic acid, 9 cis-retinoic acid, and 13-cis-retinoic acid. In addition to several described immune regulatory functions, these agents may exert their antineoplastic effects through the regulation of tumor suppressor genes such as RAR-beta2. The survival benefit provided to patients with acute promyelocytic leukemia (APL) after induction therapy with all-trans RA and the responses experienced by patients with cutaneous lesions from Kaposi's sarcoma and cutaneous T cell lymphoma treated with 9-cis RA and a selective rexinoid--bexarotene--respectively, led to their approval by the Food and Drug Administration during the last decade. As chemopreventive agents, retinoids have proven to effectively regress laryngeal papillomatosis and oral leukoplakia lesions. The ability of 13-cis-RA to prevent second primary malignancies in patients with carcinoma of the head and neck has also been demonstrated. Unfortunately, this intervention did not affect the primary tumor recurrence rates. The toxicity and efficacy of retinoids administered in combination with other biological and cytotoxic agents have also been explored in patients with renal cell carcinoma, breast cancer, myelodysplasia, prostate, cervix, and other malignancies with a broad range of reported responses. Further characterization of the molecular processes modulated by these agents will serve to better define their role in the prevention and treatment of human cancer and to tailor specific targeted therapies in combination with other compounds. Newer and more selective retinoids and rexinoids are completing phase I and phase II studies and hold promising. PMID- 12757025 TI - Peritoneal dialysis in acute renal failure. PMID- 12757026 TI - Peritoneal dialysis and infection by hepatitis B and C virus. PMID- 12757027 TI - Continuous renal replacement therapy: does technique influence electrolyte and bicarbonate control? AB - BACKGROUND AND OBJECTIVES: Different techniques of continuous renal replacement therapy (CRRT) might have different effects on electrolyte and acid-base control. The aim of this study was to determine whether continuous veno-venous hemodiafiltration (CVVHDF) or continuous veno-venous hemofiltration (CVVH) achieve better control of serum sodium, potassium and bicarbonate concentrations. DESIGN: Retrospective controlled study. SETTING: Two tertiary intensive care units. PATIENTS: Critically ill patients with acute renal failure (ARF) treated with CVVHDF (n=49) or CVVH (n=50). INTERVENTIONS: Retrieval of daily morning sodium and potassium values and arterial bicarbonate levels from computerized biochemical records before and after the initiation of CRRT for up to 2 weeks of treatment. Statistical comparison of findings. MEASUREMENTS AND RESULTS: Before treatment, abnormal (high or low) values were frequently observed for sodium (65.1% for CVVHDF vs. 80.0% for CVVH; NS), potassium (45.9% vs. 34.0%; NS), and bicarbonate (73.3% vs. 68.0%; NS). After treatment, however, CVVHDF was more likely to achieve serum sodium concentrations within the normal range (74.1% vs. 62.9%; p=0.0026). Both treatments decreased the mean serum potassium concentration over the first 48 h (p=0.0059 and p<0.0001, respectively), but there was no difference in terms of the normalization of serum potassium concentration during the entire treatment period (88.3% vs. 90.5%; NS). Both treatments increased the mean arterial bicarbonate concentration over the first 48 hours (p=0.011 and p<0.0001, respectively). However, CVVH was associated with a lower incidence of metabolic acidosis (13.8% for CVVH vs. 34.5% for CVVHDF; p<0.0001) and a higher incidence of metabolic alkalosis (38.9% vs. 1.1%; p<0.0001) during the entire treatment period. CONCLUSIONS: CRRT strategies based on different techniques have a significantly different impact on sodium and bicarbonate control. PMID- 12757028 TI - The influence of ozonated autohemotherapy on oxidative stress in hemodialyzed patients with atherosclerotic ischemia of lower limbs. AB - Ozonated autohemotherapy is used as a complementary medical approach in the treatment of vascular disorders. One of the greatest problems concerning an application of ozone in medicine is its induction of oxidative stress. The standards of ozonotherapy were elaborated recently making this treatment useful and probably non toxic. The aim of the present study was to investigate the influence of ozonated autohemotherapy on the oxidative stress extent in hemodialyzed patients, known to be particularly exposed to generation and deleterious effects of free radicals. Twelve continuously hemodialyzed subjects with atherosclerotic ischemia of the lower limbs were examined in a prospective, controlled, single blind study. Autohemotherapy with blood exposure to oxygen served as a control. The protein and lipid peroxidation products, the reduced glutathione level in red blood cells and free hemoglobin plasma concentration were measured. The study showed that ozonated autohemotherapy with ozone concentration 50 microg/ml per gram of blood induced a significant decrease in glutathione level after 9 sessions of this procedure. Therapy did not cause either the enhancement of protein and lipid peroxidation, or erythrocytes damage. It seems likely that the antioxidant defense system, part of which is glutathione, neutralizes oxidative properties of ozone in this concentration and protects against oxidative cell damage. PMID- 12757029 TI - L-carnitine consecutively administered to patients on hemodialysis improves beta cell response. AB - Eight patients with end stage renal disease (ESRD) on chronic hemodialysis (CHD) treatment were supplemented with 1 g L-carnitine intravenously (i.v.) after each dialysis session for one month. A Tolbutamide test was done and blood sugar (BS), serum C-peptide (CP) were measured at 0, 20 and 60 minutes, as well as the plasma L-carnitine level before and after treatment. Delta CP and the area under CP curve were ascertained. After L-carnitine application delta CP was significantly increased (1.33 +/- 0.63 vs. 2.24 +/- 1.0 nmol/L; p<0.05) and also the area of the stimulated secretion under the CP curve (14.93 +/- 11.11 vs. 36.88 +/- 25.36 nmol/L x 60 min.; p<0.05). The fasting BS-level was significantly lower after the treatment--3.85 +/- 0.43 vs. 4.76 +/- 1.02 mmol/L; p<0.05 and plasma L-carnitine level significantly increased (72.8 +/- 43.2 vs. 35.2 +/- 18.3 mcmol/L; p<0.05) Improving the oxidative processes in peripheral tissues, L-carnitine increases the peripheral effectiveness of insulin and relieves the overstretched beta-cell apparatus. PMID- 12757030 TI - System design of a bioartificial liver with a high performance hemodialyzer as an immunoisolator using a mathematical kinetic model. AB - We propose a new bioartificial liver (BAL) system equipped with a high performance hemodialyzer to act as an immunoisolation device. We discuss the design of the BAL system using a mathematical kinetic model with the experimentally obtained mass-transfer performances of various hemodialyzers. The mass transfer resistances of the hemodialyzers did not adversely influence the ammonia-removal and bioactive-substance-supply performances of the BAL system. A suitable hemodialyzer for the BAL system is available even at present using an engineering design. The remaining problems to be overcome before realizing clinical use of the BAL system are to increase the rate constant of the first order reaction of the BAL for ammonia metabolism and to develop a new method of blood access that can be used safely with long term reliability at a high blood flow rate (ca. 556 ml/min). PMID- 12757031 TI - Collagen expression in tissue engineered cartilage of aged human articular chondrocytes in a rotating bioreactor. AB - This study describes the culture and three-dimensional assembly of aged human articular chondrocytes under controlled oxygenation and low shear stress in a rotating-wall vessel. Chondrocytes cultured in monolayer were released and placed without any scaffold as a single cell suspension in a rotating bioreactor for 12 weeks. Samples were analyzed with immunohistochemistry, molecular biology and electron microscopy. During serial monolayer cultures chondrocytes dedifferentiated to a "fibroblast-like" structure and produced predominantly collagen type I. When these dedifferentiated cells were transferred to the rotating bioreactor, the cells showed a spontaneous aggregation and formation of solid tissue during the culture time. Expression of collagen type II and other components critical for the extracellular cartilage matrix could be detected. Transmission electron microscopy revealed a fine network of randomly distributed collagen fibrils. This rotating bioreactor proves to be a useful tool for providing an environment that enables dedifferentiated chondrocytes to redifferentiate and produce a cartilage-specific extracellular matrix. PMID- 12757032 TI - Application of Taylor vortices in hemocompatibility investigations. AB - BACKGROUND: Artificial organs, implants and extracorporeal circulation affect the physiological flow characteristics of blood as a liquid organ. These artificial systems consist of a wide variety of biomaterials with different geometries and, therefore, with their own flow properties. Secondary flow also occurs in extra- as well as in intracorporeal circulation. METHODS: In order to investigate the influence of vortical flow conditions a modified Taylor-Couette system was introduced. It consisted of two coaxial cylinders whose surfaces were the target of investigation. The annular gap was filled with donor blood shear and secondary flows were produced by rotating the inner cylinder. Platelet activation and protein adsorption were investigated as markers for thrombogenicity. RESULTS: At shear rates high enough to establish stable Taylor vortices (G > or = 550 s(-1)) significant differences between vortical Taylor flow and steady laminar flow were detected. At shear rates of G > or = 550 s(-1) laminar flow caused a significantly higher platelet drop and PF4 release when compared to Taylor vortex flow. Also protein adsorption per square unit was significantly higher for laminar flow. CONCLUSIONS: Based on the present data we conclude that vortical flow patterns lead to an accumulation of platelets and plasma proteins in the vortex center and therefore to a decreased probability of contact between platelets and material surfaces. It can be concluded that a preactivation of the platelets circulating in extracorporeal circuits can be manifested downstream in other geometrical configurations and flow conditions. PMID- 12757033 TI - Interaction between fibroblast cells and fluorinated polyimide with nano-modified surface. AB - In this study, we investigated the effect of surface nano-modification of aromatic fluorinated polyimide (6FDA-6FAP) derived from 2,2'-bis (3,4 dicarboxyphenyl) hexafluoropropane dianhydride (6FDA) and 2,2'-bis (4 aminophenyl) hexafluoropropane (6FAP) on the interaction with proteins and cells. The surface of 6FDA-6FAP was modified by surface-rubbing showed nano-ordered stripes along the rubbing direction. The rat fibroblast FR cells formed multicellular spheroids with high cell density on the rubbed surface then expressed excellent collagen production similarly in vivo, while on the non modified surface the cells formed two-dimensional monolayers and the collagen production was negligible. The modulation of cell function by the surface nano modification along with surface micro-modification may be one of the most important considerations during the design and manufacture of novel biochips or tissue engineering materials. PMID- 12757034 TI - Treatment strategies for postinfarction left ventricular free wall rupture: stabilization with peri-operative IABP and off-pump repair. AB - BACKGROUND: Perioperative management of post-infarction left ventricular free wall rupture (LVFWR) is not clearly standardized and surgical repair is the only therapeutic option. Role of off-pump surgery and stabilization with perioperative intraaortic balloon pumping (IABP) were here analysed. METHODS: Seven patients underwent surgery for LVFWR between 1990 and 2002. Clinical picture included electromechanical dissociation (3 patients) and sudden hypotension (4 patients). Except in one patient who was reanimated through femoro-femoral cardiopulmonary bypass, off-pump repair through on-lay patching technique was always performed. IABP was employed in the immediate postoperative period in five cases. RESULTS: A satisfactory hemodynamic state was restored in all cases and there were no reoperations for bleeding or rerupture. Hospital survival was 100%. One patient underwent successful surgical myocardial revascularization two months after LVFWR. Two patients died at follow-up. The survivors present with good NYHA and CCS functional classes. CONCLUSIONS: When the anatomy of the LVFWR is favourable, off-pump external patching repair proves a good choice. Postoperative IABP provides satisfactory hemodynamic support. PMID- 12757035 TI - Comparison of myocardial infarction with sequential ligation of the left anterior descending artery and its diagonal branch in dogs and sheep. AB - We report a comparison of the effects of myocardial infarction in dogs and sheep using sequential ligation of the left anterior descending artery (LAD) and its diagonal branch (DA), with hemodynamic, ultrasonographic and pathological evaluations. Five animals were used in each group. After surgical preparation, the LAD was ligated at a point approximately 40% of the distance from the apex to the base of the heart, and after one hour, the DA was ligated at the same level. Hemodynamic and ultrasonographic measurements were performed preligation, 30 minutes after LAD ligation, and 1 hour after DA ligation. As a control, two animals in each group were used for the simultaneous ligation of the LAD and the DA. Two months after the coronary ligation, the animals were evaluated as previously, and killed for postmortem examination of their hearts. All seven animals in the dog group survived the experimental procedures, while in the sheep group only animals with sequential ligation of the LAD and DA survived. Statistically significant decreases in systemic arterial blood pressure and cardiac output, and an increase in the pulmonary artery capillary wedge pressure (PACWP) were observed one hour after sequential ligation of the LAD and its DA in the sheep, while only systemic arterial pressures decreased in the dog. Ultrasonographic analyses demonstrated variable degrees of anteroseptal dyskinesia and akinesia in all sheep, but in no dogs. Data two months after coronary artery ligation showed significant increases in central venous pressure, pulmonary artery pressure, and PACWP in the sheep, but not in the dog. Left ventricular end-diastolic dimension and left ventricular end-systolic dimension in ultrasonographic studies were also increased only in the sheep. Pathologically, the well-demarcated thin-walled transmural anteroseptal infarcts with chamber enlargement were clearly seen in all specimens of sheep, and only mild-to-moderate chamber enlargements with endocardial fibrosis were observed in the dog hearts. In conclusion, this study confirms that the dog is not a suitable model for myocardial infarction with failure by coronary artery ligation despite negligent operative mortality, when compared directly with an ovine model. PMID- 12757036 TI - Levofloxacin elimination during albumin dialysis. PMID- 12757037 TI - Severe acute respiratory syndrome: a lesson in infectious disease. PMID- 12757038 TI - Asthma, a disease of rich and poor. PMID- 12757039 TI - A prioritised research agenda for DOTS-Plus for multidrug-resistant tuberculosis (MDR-TB). PMID- 12757040 TI - Increasing prevalence of asthma in UK primary care during the 1990s. AB - OBJECTIVE: To estimate recent prevalence trends of physician-diagnosed asthma in primary care in the UK, and to test the hypothesis that the asthma epidemic in the UK peaked in the mid-1990s and is currently declining. METHODS: A retrospective cohort of asthma patients was obtained from the General Practice Research Database (GPRD). From January 1990 to February 1999, asthmatics were followed up to death, censoring or mention of chronic obstructive pulmonary disease (COPD) in their clinical record. Prevalence rates of ever and managed asthma were obtained by sex, age and calendar year. RESULTS AND CONCLUSION: From 1990 to 1998, annual prevalence rates of managed physician-diagnosed asthma in women rose from 3.01% (95%CI 2.99-3.03) to 5.14% (95%CI 5.10-5.18), and in men from 3.44% (95%CI 3.41-3.46) to 5.06% (95 %CI 5.02-5.10) (P for trend <0.01 in both). In 1998, prevalence rates of managed asthma in children aged 5-14 affected 7.86% (95%CI 7.71-8.00) of girls and 10.30% (95%CI 10.15-10.47) of boys. Increasing prevalence rates in adult asthma (maximum 4.11% in 1998, 95%CI 4.03 4.19) and elderly asthma (maximum 3.37% in 1998, 95%CI 3.29-3.46) were observed as well in 1998. The study shows that the burden of asthma in UK primary care during the 1990s was still increasing. PMID- 12757041 TI - Lack of essential asthma medications in primary care centres in Kuwait. AB - SETTING: Asthma is a common chronic disease that can affect any age group. Available, affordable and effective asthma medications are essential to achieve good asthma control. OBJECTIVES: To determine the availability of essential asthma medications in primary care centres and the cost of a year's supply in Kuwait. METHODS: A telephone survey was conducted in 36 centres. The cost of asthma medication was calculated based on estimated annual needs for the different grades of asthma severity. RESULTS: None of the centres surveyed had adequate asthma medication: 12 (33%) had just adequate and 24 (67%) had below adequate supplies. All had intravenous hydrocortisone and nebulised bronchodilators, but none had high-dose inhaled steroids; 24 (67%) had neither oral steroids nor salbutamol inhalers. All had oral bronchodilators, aminophylline and theophylline. Family medical centres had more asthma medications. The annual cost of treating a case of moderate asthma in Kuwait is 562 US dollars. CONCLUSION: Most of the primary care centres in Kuwait lack essential asthma medications. This creates over-reliance on hospital emergency departments and affects asthma management. PMID- 12757042 TI - Adult-to-child transmission of tuberculosis: household or community contact? AB - SETTING: If a child develops tuberculosis, it is assumed that the source was an adult infectious case, usually living in the same house. Restriction fragment length polymorphism (RFLP) was used in this study to establish transmission from source cases to children. DESIGN: Adult and child tuberculosis cases were prospectively identified from 1993 to 1998 and cultures of Mycobacterium tuberculosis collected. Interviews and RFLP analysis of M. tuberculosis strains were performed to establish epidemiological links and to confirm household transmission. RESULTS: Tuberculosis was confirmed by culture in 1139 (91%) of 1291 adults and 65 (16%) of 417 children. Due to problems in recovering specimens or extracting DNA, RFLP analysis was done in 832 adults and 35 children: 19 (54%) children had household members identified with tuberculosis, 12 with the same strain as the child. Twenty-nine (83%) strains from children formed part of community clusters, but definite contact with source cases was established in only 15. CONCLUSION: The presence of an adult with infectious tuberculosis in the same house as a child with tuberculosis does not necessarily imply adult-to-child transmission. Young children may be infected in the community or in the household. These findings have implications for contact tracing and treatment strategies in high incidence areas. PMID- 12757043 TI - Factors associated with time to sputum smear conversion in active pulmonary tuberculosis. AB - OBJECTIVE: To ascertain the factors affecting the time between the initiation of treatment and obtaining three negative sputum smears. DESIGN: In a study of 109 patients with pulmonary tuberculosis, the main variable was the period during which the patients had sputum smears once treatment was initiated. Multivariate analysis (multiple linear regression) was performed to document those variables independently associated with time to conversion. RESULTS: The patients had positive smears for a mean of 28.63 days. The most frequent radiographic pattern was cavitary disease (36.7%). HIV co-infection was present in 38.5% of the patients. HIV-infected patients showed a cavitation pattern in only 9.6% vs 52.2% of patients without HIV infection (P < 0.001). The variables that showed a statistically significant and independent relationship with the time to sputum smear conversion were pulmonary radiographic pattern, age and erythrocyte sedimentation rate (ESR). CONCLUSIONS: ESR, age and the presence of cavitary disease seem to be factors associated with a longer time to sputum smear conversion in patients with active pulmonary tuberculosis. However, HIV co infection is associated with a shorter time to sputum conversion. A key factor is therefore the presence or not of cavitation, independently of HIV infection. PMID- 12757044 TI - Systematic restaining of sputum smears for quality control is useful in Burundi. AB - SETTING: Routine tuberculosis control services in Burundi. OBJECTIVE: To determine whether systematic restaining of sputum smears for acid-fast bacilli (AFB) prior to rechecking quality assessment is necessary. DESIGN: Blinded rechecking of peripheral routine smears, including a second control of discordants, before and after restaining. RESULTS: Without restaining, 10/825 (1.2%) negative, and 59/189 (31.2%) positive results were declared false. After restaining, there were 34 (4.1%) false negatives and 13 (6.9%) false positives, both highly significant changes. Before restaining, quantification of positive smears was usually considered too high, while after restaining 41 out of 42 positives were found to have too low readings. CONCLUSIONS: Despite mild climatic conditions in Burundi, restaining all slides before rechecking revealed an unrecognised, widespread problem of false negatives, rather than false positives. This indicated the need for critical re-appraisal of the standard procedure using cold staining, rather than re-training, as would have been inferred from results without restaining. Systematic restaining of all slides prior to rechecking may be more widely needed in National Tuberculosis Programmes to cover all possible serious causes of error. Cold staining should be avoided in field programmes since its performance is easily affected by frequently encountered adverse factors. PMID- 12757045 TI - The effect of decentralisation on tuberculosis services in three states of Sudan. AB - SETTING: Referral hospitals and primary health care (PHC) facilities in Khartoum, Red Sea and Gadaref States. OBJECTIVES: To measure the effect of the decentralisation of the tuberculosis (TB) services on the clinical profile and treatment outcome of tuberculosis. DESIGN: A cohort study of case detection and treatment outcome using information routinely collected comparing patients attending PHC facilities and referral hospitals in selected locations in Sudan. RESULTS: Two-thirds of all TB patients were diagnosed in referral hospitals and one-third in PHC facilities. In PHC facilities, women represented 46% of notified cases, compared to 37.9% in referral hospitals (OR 1.398, 95%CI 1.343-1.455). Older age groups were more likely to prefer PHC facilities to referral hospitals. In referral hospitals, 38% were cured, 29.3% completed treatment without smear examination and 17.3% defaulted, while in PHC facilities 58% were cured, 17.8% completed treatment without smear examination and 11.6% defaulted. CONCLUSION: PHC facilities provide care for a higher proportion of women and older age groups of tuberculosis patients, suggesting a higher level of accessibility for these groups. A higher cure rate and a lower default rate were noted in PHC facilities, possibly reflecting better conditions for directly observed treatment and follow up. PMID- 12757046 TI - Differential decline in tuberculosis incidence among US- and non-US-born persons in New York City. AB - SETTING: A large urban tuberculosis control program. OBJECTIVES: To examine changes in tuberculosis incidence and characteristics of cases in New York City (NYC), and assess the epidemiology of tuberculosis among non-US-born persons. DESIGN: Tuberculosis surveillance data (1995-1999) for NYC were analyzed. RESULTS: Tuberculosis incidence decreased by 56.6% in US-born and 19.6% in non-US born persons (age-adjusted) over the study period. The decline in tuberculosis incidence among US-born persons was more substantial in the first half of the study period (23-24%) than in the second half (13-15%). The greatest decline in incidence was among US-born Hispanic or Black males aged 25-64. However, although there was an overall decline in incidence among non-US-born persons, there was no significant change in any sex or racial/ethnic subgroup. The percent of multidrug resistant (MDR) cases among non-US-born patients remained stable, but recent arrivals accounted for 79% of non-US-born MDR-TB patients in 1999, a significant increase from 16% in 1997. CONCLUSIONS: Continuing current tuberculosis control efforts and treatment of immigrants with latent tuberculosis infection are of highest priority for reducing incident cases in NYC. Global collaboration towards earlier detection and treatment of active tuberculosis cases in high incidence countries is also essential. PMID- 12757047 TI - TB status among Kosovar refugees. AB - DESIGN: In the spring of 1999, 864 Kosovars were directly airborne from refugee camps in Macedonia to a refugee camp in Kristiansand, Norway; 800 were examined according to official Norwegian TB screening procedures with X-ray (if more than 15 years of age) and tuberculin test shortly after arrival. RESULTS: The mean (SD) age was 29.2 (18.7) years of age and 29% were aged under 15 years; 79% of the refugees had escaped from urban areas in Kosovo, and 75% had an identifiable BCG scar. Among those with BCG scar, increasing age and male sex were associated with a significant tuberculin reaction: 20% had tuberculin reactions indicating latent TB infection, while 40% had negative tuberculin reactions. Approximately 4% of the refugees aged over 15 had abnormal chest X-rays, predictive of an enhanced tuberculin reaction. Four refugees had X-ray findings compatible with active TB and were treated with standard four-drug chemotherapy. CONCLUSION: The Kosovar refugees had a high incidence of active tuberculosis (50/100,000). A fifth of the BCG-vaccinated refugees needed careful follow-up to monitor possible progress from latent to active TB infection after immigration, while one in seven non-BCG-vaccinated refugees had tuberculin skin reactions compatible with latent TB infection. PMID- 12757048 TI - Public-private mix for improved TB control in Ho Chi Minh City, Vietnam: an assessment of its impact on case detection. AB - SETTING: Ho Chi Minh City (HCMC), Vietnam. OBJECTIVE: To assess the impact on case detection of a public-private mix (PPM) project linking private providers (PPs) to the National Tuberculosis Programme (NTP). METHOD: Nine-month monitoring of referral and diagnostic data recorded in new referral forms and treatment cards for PPs and upgraded NTP registers. RESULTS: A total of 1549 TB suspects were referred, of whom 1090 (70%) actually went to the NTP for sputum examination. A total of 569 cases were detected through referrals or notification, of whom 45% were new sputum smear-positive cases. The case detection of new sputum smear-positive cases in PPM districts increased by 18% (21/100,000, 95%CI 0-42) compared to the previous year, while a slight decrease occurred in control districts. In HCMC as a whole, case detection increased by 7% (7/100,000, 95%CI 2-11/100,000). Among sputum smear-positive cases detected in NTP through referrals from PPs, 58% defaulted before initiating treatment. CONCLUSIONS: The tendency towards increased case detection associated with this PPM indicates a potential for utilising PPs to improve case detection. However, the NTP and PPs should jointly address the problem of initial default before considering expansion of this PPM model. PMID- 12757049 TI - Resistance to anti-tuberculosis drugs among smear-positive cases in Thai prisons 2 years after the implementation of the DOTS strategy. AB - SETTING: Three prisons in Bangkok and vicinity, Thailand. OBJECTIVE: To examine the prevalence of drug-resistant tuberculosis among smear-positive cases in Thai prisons 2 years after the implementation of the DOTS strategy, and to identify factors associated with resistance to anti-tuberculosis drugs. METHODS: In a cross-sectional study, 154 consecutive tuberculosis patients with at least one positive sputum smear and at least one positive sputum culture registered between 1 May and 31 October 2000 were enrolled. Drug susceptibility testing was performed by the Ministry of Public Health Tuberculosis Division. Patient characteristics were obtained by face-to-face interview. RESULTS: Resistance to at least one drug was found in 50.6% of the subjects, including multidrug resistant tuberculosis (MDR-TB) in 19.5%. The proportion of resistance to any anti-tuberculosis drug in prisons A, B and C was respectively 52.7%, 37.8% and 61.5%. The only factor significantly associated with resistance to at least one drug (P = 0.011) and MDR-TB (P < 0.001) was a history of previous tuberculosis treatment. CONCLUSION: After 2 years of the DOTS strategy, resistance to anti tuberculosis drugs, an indicator of the quality of tuberculosis treatment, was found to be high. The DOTS strategy currently used in Thai prisons should be reviewed, in order to reduce and prevent drug-resistant tuberculosis. PMID- 12757051 TI - How can detection of infectious tuberculosis be improved? Experience in the Somali region of Ethiopia. AB - In early 1999, 48% of pulmonary tuberculosis (PTB) cases detected in the Somali region of Ethiopia were smear-positive. Actions at the laboratory level and peer review of smear-negative PTB diagnoses were proposed. Clinicians knew, but did not adhere to, the algorithm recommended by the National Tuberculosis Programme for these diagnoses, partly due to the costs involved to patients. Challenging clinicians, in a non-threatening way, to become more clinically rigorous proved successful, and the proportion of smear-positive PTB increased to 65%. Operational research is needed to assess the feasibility of these widely recommended smear-negative PTB diagnosis guidelines. PMID- 12757050 TI - Factors associated with humoral response to ESAT-6, 38 kDa and 14 kDa in patients with a spectrum of tuberculosis. AB - SETTING: Tertiary care chest hospital in Montreal, Canada, where the average annual incidence of TB is 10/100,000 population. OBJECTIVES: To evaluate the clinical correlates of humoral response to three Mycobacterium tuberculosis antigens. METHODS: Humoral response to three M. tuberculosis antigens, 38 kDa, 14 kDa and ESAT-6, was measured with ELISA in patients with a spectrum of TB-related conditions. The association of positive tests for each antigen, defined with receiver operator characteristics (ROC) analysis, and patient characteristics was assessed in multivariate regression. RESULTS: A total of 383 patients underwent serologic testing. In multivariate analysis, humoral response to 38 kDa was associated with active disease, response to 14 kDa was associated with inactive TB and female sex, and response to ESAT-6 with inactive TB, female sex, prior contact with TB, and recent arrival in Canada from high prevalence countries. CONCLUSIONS: Response to the 38 kDa antigen was associated with current active disease, and was very different from response to the 14 kDa and ESAT-6 antigens. These latter two antigens were associated with risk factors for future active, but not current disease, suggesting that they might be useful to identify persons with higher risk of reactivation of latent TB. PMID- 12757052 TI - Tuberculosis in health care workers in a central hospital in Malawi. AB - A retrospective study was carried out at Lilongwe Central Hospital, Malawi, to determine 1) the tuberculosis case notification rate in health care workers (HCW) in 2001, and 2) whether NTP guidelines were adhered to in diagnosing TB. Of 571 HCWs, 33 (6%) were notified with TB in 2001, giving a TB case notification rate of 5780/100,000. Patient attendants had higher rates of TB than nurses, ward attendants and doctors, but otherwise there were no significant differences between the HCW categories. NTP diagnostic guidelines were not properly followed, particularly in diagnosing smear-negative TB; in HCWs with smear-negative PTB, 64% had no sputum smears examined. The rate of TB in HCWs in a central hospital in Malawi was high, although lack of adherence to guidelines might have resulted in overdiagnosis. PMID- 12757053 TI - Para-aminosalicylic acid (PAS) desensitization review in a case of multidrug resistant pulmonary tuberculosis. AB - SETTING: Tertiary care hospital in the Upper Midwest, United States. OBJECTIVE: Rapid desensitization to para-aminosalicylic acid (PAS) in a patient with previous hypersensitivity reaction and a review of published PAS desensitization protocols. DESIGN: Composition and implementation of a short-course PAS desensitization protocol for a 34-year-old woman with multidrug-resistant (MDR) pulmonary tuberculosis, incorporating published experiences of PAS desensitization over the past 50 years. RESULTS: We composed a protocol and successfully desensitized our patient to PAS (Paser granules). By starting with a low dose (50 mg), then doubling the PAS dose on each successive day, our patient was able to tolerate full dose in 1 week. No steroids were required and no adverse reactions were encountered. Previous published PAS desensitization protocols used starting doses of 10-500 mg, desensitization time ranges from 7 to 54 days and commonly used steroids or corticotropin. CONCLUSION: Rapid desensitization to PAS can be successfully conducted within 1 week without the use of steroids or corticotropin. Given the limited number of drugs available for many patients with MDR-TB, desensitization to PAS is a valid alternative to drug discontinuation for patients with hypersensitivity reactions. PMID- 12757054 TI - Disseminated Mycobacterium avium complex (DMAC) in an immunocompetent adult. AB - A female patient with multiple osteomyelitis and pulmonary Mycobacterium avium disease visited an orthopaedic clinic with back pain. Systemic bone scan showed multiple sites of increased radioactivity in the vertebral bodies, right scapula, femurs and ribs. M. avium was isolated from sputum and a sample aspirated from the right scapula. The route of infection was unknown as there was no history of trauma or surgery. HIV testing was negative. As there was no underlying immunological disease she was diagnosed as disseminated M. avium complex (DMAC) disease in an immunocompetent adult. Cytokine production on several stimuli from peripheral blood mononuclear cells was similar to that in pulmonary M. avium patients. Sequence analysis of IFN-gamma receptor revealed no nucleotide substitution. We detected serotypes 1, 2 and 4 from mycobacteria cultured from the right scapula, and conclude that this case could be the result of undetected immune deficiency and/or unrecognised virulence of the infecting isolate. PMID- 12757055 TI - Disseminated Mycobacterium genavense in an HIV-infected patient from South America. PMID- 12757056 TI - Pleural effusion due to Mycobacterium gordonae infection. PMID- 12757057 TI - Mycobacterium tuberculosis presenting as an infected pulmonary bulla. PMID- 12757058 TI - The efficacy and safety of oral sildenafil in Thai men with erectile dysfunction: a randomized, double-blind, placebo controlled, flexible-dose study. AB - OBJECTIVE: To evaluate the efficacy and safety of sildenafil citrate (Viagra) in a randomized, double-blind, placebo-controlled, flexible-dose study in Thai men with erectile dysfunction of broad-spectrum etiology and more than 6 months' duration. MATERIAL AND METHOD: 125 patients aged 26 to 77 years were randomized at 4 centers in Thailand to receive either sildenafil citrate (50 mg initially, increased if necessary up to 100 mg or decreased to 25 mg depending on efficacy and/or tolerability) (n = 63) or a matching placebo (n = 62) taken on an 'as needed' basis approximately 1 hour prior to anticipated sexual activity for a period of 12 weeks. Efficacy was assessed by the patients' responses to the 15 question International Index of Erectile Function (IIEF), to questions on the event log of sexual activity, and to the global efficacy assessment question concerning improvement in erections. RESULTS: At the conclusion of the study, both the primary efficacy variables relating to the achievement and maintenance of erections sufficient for sexual intercourse and the secondary efficacy variables, which included the 5 separate domains of sexual functioning of the IIEF, the percentage of successful attempts at sexual intercourse, and the global efficacy assessment question concerning improvement in erections, were all significantly improved statistically by sildenafil in comparison with placebo except in the sexual desire domain which showed no difference. The percentage of successful attempts at sexual intercourse in the sildenafil group was 66.16 per cent while in the placebo group it was 33.05 per cent. The percentage of global efficacy assessment was improved in the sildenafil group by 82.5 per cent compared to 36.1 per cent in the placebo group. Adverse events considered treatment-related occurred in 19 patients (30.2%) receiving sildenafil and 7 (11.3%) receiving placebo. The most common adverse events with sildenafil were vasodilatation (flushing), headache, and dizziness, which occurred in 14.3 per cent, 6.3 per cent, and 6.3 per cent of patients respectively. All events were mild in nature. CONCLUSIONS: Sildenafil is a safe and effective treatment for erectile dysfunction of broad-spectrum etiology in Thai men. Its efficacy appears similar to that reported in other studies in Western populations. PMID- 12757059 TI - Clinical indications for penetrating keratoplasty in Maharaj Nakorn Chiang Mai Hospital, 1996-1999. AB - PURPOSE: To determine the leading indication for penetrating keratoplasty. MATERIAL AND METHOD: The authors retrospectively performed a chart review of the hospital records of all patients who underwent penetrating keratoplasty at Maharaj Nakorn Chiang Mai Hospital between January 1996 and December 1999. RESULTS: A total of 45 corneal transplants were performed. The leading indications for penetrating keratoplasty, in order of decreasing frequency, were bullous keratopathy (28.9%), corneal scar (22.2%), corneal dystrophy and degeneration (20.0%), corneal ulcer (17.8%), regraft (8.9%), and trauma (2.2%). CONCLUSION: Bullous keratopathy was the leading indication for penetrating keratoplasty at Maharaj Nakorn Chiang Mai Hospital from 1996 to 1999, followed by corneal scar. The major cause of bullous keratopathy was associated with posterior chamber intraocular lens implantation. PMID- 12757061 TI - Peri-operative factors predicting the outcome of hepatic porto-enterostomy in infants with biliary atresia. AB - INTRODUCTION: Without hepatic transplantation, hepatic porto-enterostomy is the only definitive surgical therapy for infants with biliary atresia. Unfortunately, clearance of jaundice by the procedure is not promising. Pre-operative data that may predict the outcome is of great value in the selection of surgical candidates. Early post-operative determinants of outcome also help in follow-up planning. OBJECTIVE: To determine peri-operative factors influencing jaundice clearance after hepatic porto-enterostomy in infants with biliary atresia. PATIENTS AND METHOD: Clinical and laboratory data of pediatric patients undergoing hepatic porto-enterostomy in Songklanagarind Hospital from 1988 to 2001 were reviewed regarding age at operation, clinical presentation, gross pathology of bile duct atresia, liver function profiles and changes after the procedure, liver pathology and post-operative ascending cholangitis. Univariate comparison followed by multivariate logistic regression analysis was analyzed against the clearance of jaundice. Statistical analysis was aided by the Stata 7.0 program. Statistical significance was set at p-value less than 0.05. RESULTS: There were 62 infants operated on during the thirteen-year period. Four cases of operative death and a case lost to follow-up before the second post-operative month was excluded. The median age at the operation was 78 days (34-326 days). Twenty-four cases (42.1%) presented with signs of portal hypertension. After the operation, 19 cases (33.4%) were jaundice free, 6 cases (10.5%) had fair clearance and 32 cases (56.1%) had a poor result. Univariate analysis revealed an association between age at surgery and jaundice clearance. Post-operative stool color and decline of total bilirubin level at one month after surgery were significantly correlated with the outcome (p < 0.01). Cholangitis within the first post-operative month significantly had an adverse effect on the short-term survival probability. Multivariate analysis showed an independent association of jaundice clearance with age at surgery and type of bile duct atresia. CONCLUSION: Age of the infants younger than 60 days and type I of bile duct atresia were the key determinants of successful hepatic porto-enterostomy. Early cholangitis was an accelerator of progressive cirrhosis. Stool color and bilirubin level at one month after surgery can be used as predictors of jaundice clearance. PMID- 12757060 TI - Risk index for predicting complications and prognosis in Thai patients with neutropenia and fever. AB - BACKGROUND: New strategies in the treatment of febrile neutropenic patients have been proposed during the past decade. It is more and more widely accepted that febrile neutropenic patients are a heterogeneous population and they have varying risks for complications and death. However, most of the data have been collected from patients in Western countries. The purpose of the study was to identify types of infection and etiologic organisms in febrile neutropenic patients at Siriraj Hospital, Thailand, and also to develop a prediction model in order to identify patients who are expected to have a favorable outcome or a low-risk subset. METHOD: The medical records of chemotherapy-induced neutropenic patients with fever hospitalized at Siriraj Hospital, Thailand, from January 1999 to December 2000 were analyzed. Data included patient characteristics, epidemiological data and the potential factors at the onset of fever for predicting patient outcome. A scoring system for predicting patients with favorable outcome was developed. The scoring system developed from this study was compared with a previously used scoring system. RESULTS: Of 220 patients with 267 febrile neutropenic episodes, 71.8 per cent had hematologic malignancies and 28.2 per cent had solid tumors. Bacteremia was found in 61 episodes (22.8%) and gram negative bacilli were the most common causative organism in bacteremia (88.6%). Overall mortality was 17.7 per cent. Multivariate analysis revealed that the factors predicting outcome were burden of illness, control of cancer, duration of neutropenia and dehydration. The scoring system developed from this set of data revealed that a score > or = 16 identified patients with a favorable outcome with a specificity of 90.2 per cent, sensitivity of 76.6 per cent and positive predictive value of 85.4 per cent. CONCLUSION: The causative organisms of bacterial infections in febrile neutropenic patients in Thailand are still gram negative bacteria. The locally developed risk index has a fair accuracy to identify patients with favorable outcome and may be used to identify patients suitable for less aggressive treatment strategies. PMID- 12757062 TI - Suprafibrous injection with corticosteroid in de Quervain's disease. AB - Between February 1996 and August 2001, 115 suprafibrous injections with corticosteroid were performed on 103 patients. Initial satisfactory result was found in 105 wrists (91.30%). The average duration of follow-up was 34 months. Seventy-one wrists (61.74%) had no recurrence of the symptom. The average pain free interval in the patient who had recurrence of the symptom after having an initial satisfactory result was five months. The success rate following one to three suprafibrous injections was 77.39 per cent. Suprafibrous injection is technically easier and has less risk of intratendinous injection than intrasynovial injection. The accuracy of injection and anatomical variation of the first extensor compartment of the wrist affect the result of intrasynovial injection but will not affect the result of suprafibrous injection in de Quervain's disease. PMID- 12757063 TI - Optic neuritis: characteristics and visual outcome. AB - OBJECTIVE: To determine clinical characteristics of patients with optic neuritis and visual outcome after intravenous methylprednisolone treatment. METHOD: A total of 81 patients with optic neuritis were reviewed retrospectively with regard to their clinical characteristics by dividing into two groups as follows: group I had isolated optic neuritis and group II had optic neuritis with demyelinative disease. The visual outcome in these patients before and after intravenous methylprednisolone treatment was analyzed. RESULTS: Of 81 patients with optic neuritis, 63 patients (77.8%) had isolated optic neuritis and 18 (22.2%) patients were optic neuritis with demyelinative disease. The ages of the patients ranged from 16 to 59 years (mean = 35.3 years) in patients with isolated optic neuritis and from 16 to 73 years of age (mean = 35.8 years) in patients with optic neuritis with demyelinative disease. After treatment, 45 patients (52 eyes) with isolated optic neuritis and 14 patients (25 eyes) with optic neuritis with demyelinative disease who were followed-up for more than 10 days were studied. After treatment, 60 per cent of the isolated optic neuritis patients and 24 per cent of the optic neuritis patients with demyelinative disease had a visual acuity of 6/12 or better respectively. The isolated optic neuritis who had an onset interval to treatment of less than 8 days had a visual acuity better than 6/9 in 75 per cent. CONCLUSION: The final visual outcome in patients with isolated optic neuritis who received earlier treatment was better than those who received treatment later. PMID- 12757064 TI - The efficacy of ginger in prevention of post-operative nausea and vomiting after outpatient gynecological laparoscopy. AB - OBJECTIVE: To study the efficacy of ginger in prevention of nausea and vomiting after outpatient gynecological laparoscopy. STUDY DESIGN: Double blind randomized controlled trial. SETTING: Department of Obstetrics and Gynecology, Thammasat Hospital, Faculty of Medicine, Thammasat University. MATERIAL AND METHOD: From January, 2001-December, 2001, 80 patients who underwent outpatient gynecological laparoscopy were randomly allocated into group A (n = 40) and group B (n = 40). The patients in group A received 2 capsules of ginger (1 capsule contain 0.5 g of ginger powder) 1 h before the procedure while the patients in group B received the placebo. The visual analogue nausea scores (VANS) and vomiting times were evaluated at 2, 4 and 24 hours after operation. RESULT: There was a significant difference in the incidence of the nausea between group A [12 (30%)] and group B [23 (57.50%)]. The VANS was lower in group A than in group B at 2 and 4 hours (p < 0.05). No difference of VANS at 24 hours was found in both groups. Incidence and frequency of vomiting in group A were lower than group B but there were not statistically different. CONCLUSION: From our data, ginger is effective in prevention of nausea after outpatient gynecological laparoscopy. PMID- 12757065 TI - New visual acuity chart: modification for clinical practice and research. AB - The authors created numeric optotypes by steps following the standard Sloan letters and developed a visual acuity test chart using Logarithm of the Minimal Angle of Resolution (Log MAR) such as the Early Treatment Diabetic Retinopathy Study (ETDRS) chart. Legibility of new numeric optotypes was presented in per cent of correct response at threshold. Only seven numbers (0, 2, 3, 5, 6, 8, 9) were used as optotypes and the average of per cent of correct response at threshold equal to 80.7 which was slightly less than standard Sloan letters (82%). The comparison between results of visual acuity level from the new chart and ETDRS showed that the ETDRS chart was slightly more legible than the new chart. It can be used universally especially with Thai people for clinical practice and research. PMID- 12757066 TI - A controlled trial of a new treatment for galactocele. AB - BACKGROUND: Needle aspiration, followed by excision should it recur, is the standard method of treating galactocele. Villagers in Northeast Thailand traditionally treat galactocele by probing the obstructed duct with double strands of pleated human hair. The aim of the study was to mimic this method in order to scientifically assess its effectiveness. PATIENTS AND METHOD: Sixteen patients were consecutively enrolled between 1995 and 2001. They elected either standard needle aspiration (Group A) or treatment by 6-0 nylon probing (Group B). The results were compared using the Fisher's exact and Mann-Whitney tests at p value < 0.05. RESULTS: The two groups were similar regarding the children's age, first para, mass size, and duration of symptoms, but patients in the aspiration group were considerably younger than the nylon probing group. Both methods reduced the symptoms completely. Pain from treatment was reported by all patients in the aspiration method while there were none in the nylon probing method (p < 0.001). The aspiration method took 14.8 minutes less time than the nylon probing method (p < 0.001). Recurrence was found in 2 out of 5 patients in the aspiration method, whereas there was none in the 11 patients with the nylon probing method (difference = 40%; 95% CI: -3% to 83%; p = 0.083). CONCLUSIONS: The new treatment of galactocele by nylon probing took longer than aspiration but removed the protein plug that caused obstruction of the duct without pain and had a tendency to reduce the recurrence rate. PMID- 12757067 TI - Spinal morphine for post-operative analgesia after lumbar laminectomy with fusion. AB - BACKGROUND: Intrathecal administration of preservative free morphine (spinal morphine) provides excellent post-operative analgesia. Since the dura is readily accessible by the surgeon during lumbar spinal surgery, it would be convenient and attractive to administer morphine into the spinal space to provide adequate post-operative analgesia in these patients. METHOD: A prospective randomized controlled study evaluated the post-operative analgesic effect of spinal morphine after lumbar laminectomy with fusion. Forty patients were randomly allocated to two groups, morphine (MO) or normal saline (NSS). Morphine 0.3 mg in normal saline 0.3 ml or normal saline 0.3 ml was injected into the dural sac under direct visualization before closing the wound. An intravenous PCA morphine device was provided for post-operative pain relief. RESULTS: Median visual analog scale (VAS) pain scores were lower in the MO group at 2, 4, 24 and 48 h after surgery (1, 1, 2.75 and 1.5 cm in the MO group vs 4.25, 4.25, 5 and 4 cm in the NSS group) (p < 0.05). The time to first patient control analgesia (PCA) demand was delayed in the MO group (131.7 min vs 29.6 min) (p < 0.05). The cumulative doses of PCA morphine consumption were lower in the MO group in the first 24 h and 24 48 h (13.7 and 15.9 mg vs 41.3 mg and 27.1 mg) (p < 0.001). The incidence of pruritus was higher in the MO group in 24 h and 24-48 h (45%, and 45% vs 5% and 10%) (p < 0.05). The incidence and severity of nausea, vomiting and sedation were not different. No patient developed respiratory depression or postdural puncture headache (PDPH). The patients' satisfaction with post-operative pain management was 100 per cent in the MO group and 85 per cent in the NSS group. CONCLUSION: Spinal morphine improved post-operative pain relief after lumbar laminectomy. PMID- 12757068 TI - The midpedicular and interpedicular distance of thoracolumbar vertebrae. AB - OBJECTIVES: 1. To introduce the measurement of the midpedicular distance (MPD) as an alternative to the interpedicular distance (IPD). 2. To measure the IPD and MPD of T11, T12, L1 and L2 vertebrae on anteroposterior (AP) plain film of the normal spine. 3. To study the relationship of the IPD and MPD of T11 to L2 of the normal spine. STUDY DESIGN: Thoracolumbar AP plain film of 89 subjects (39 males, 50 females) with an average age of 47.6 years (range 21-78 years) from the roentgenographic files were included. Both the IPD and MPD of T11, T12, L1 and L2 were measured by two observers. The mean difference of IPD and MPD at these four levels were compared by using a one-way ANOVA. The relationship of the IPD and MPD measured from the T11 to L2 levels were evaluated using the simple linear regression model. RESULTS: The mean IPD was progressively wider (p = 0.000) from the T11 to L1 level, but no significant difference (p = 0.308) was found between the mean IPDs of L1 and L2. The mean MPD of each level was significantly different from the others (p = 0.000) except the mean MPDs of T12 and L1 (p = 1.000). Both the IPD and MPD had a statistically significant linear relationship with the level of the vertebrae from T11 to L2 (p = 0.000) with the coefficients of determination (R2) of 0.39 and 0.28, respectively. CONCLUSION: More care should be taken in clinical practice when measuring the IPD of a fractured vertebra relative to those of adjacent lower levels in order to determine whether or not widening has occurred, especially IPD L1 vis-a-vis IPD L2. MPD measurement has no advantage over the IPD measurement but is a useful alternative when a comparison of the pedicular distance of L1 and L2 is needed. PMID- 12757069 TI - Preliminary study on somatic cell nuclear transfer in rabbits in Thailand. AB - The objective of the study was to develop the somatic nuclear transfer technique by using rabbits as the model. The oocyte recipients aged 16 h post coitus were collected surgically from 20 superovulated rabbit doe with 28 and 40 mg Follicle Stimulating Hormone (FSH) after mating with a vasectomized male. The metaphase II plate and 1st polar body of oocyte was later aspirated by enucleated micropipette under an inverted microscope. A single donor cell; cumulus cell or cultured or frozen fibroblast cell from passage 1 to 9 were transferred to enucleated oocyte and fused with triple DC pulses, 3.2 kv, 20 micros. The fused embryos were cultivated in TCM 199 NaHCO3 + 10 per cent fetal calf serum (FCS) for 4 days. The cleavage rate (2-cell stage) was 37.2 per cent (32/86) from eight experiments, and 18.8 per cent (6/32) developed to the early morula stage. This study also indicated that the enucleation pipette and the somatic cell type influenced the success. PMID- 12757070 TI - A knee model for arthrocentesis simulation. AB - OBJECTIVES: To produce a knee model for medical students and residents to practice knee aspiration and intra-articular injection. MATERIAL AND METHOD: The model was made of plastic, rubber and silicone that included the lower third of the femur, the upper third of the tibia and the patella. They were fixed on 2 plastic boxes in the anatomical position and the boxes were connected together with 2 small hinge joints. A rubber bag was made in the pattern of synovial space of the knee. Quadriceps, anterior muscles of the leg and patellar tendon were also presented. The model was covered with silicon sheet, representing the skin. Water was used to fill up the synovial bag to simulate joint aspiration via supero-lateral approach with the knee in extension. The model was appraised by 30 medical students, 26 orthopedic residents and 10 orthopedic staff in terms of size, anatomy, physical examination, feeling during aspiration, need of the model in education and commercial use. RESULTS: Most of the medical students, residents and staff (80-90%) were satisfied with the model and rated it as good to very good teaching media. However, the model should come out in different sizes and the synovial bag should be modified to improve the ballotment test. Mass production of the model should be done. CONCLUSION: The knee model is an acceptable teaching model for arthrocentesis simulation with affordable cost. PMID- 12757071 TI - The teaching of ethics. PMID- 12757072 TI - The efficacy of fluconazole 600 mg/day versus itraconazole 600 mg/day as consolidation therapy of cryptococcal meningitis in AIDS patients. AB - Cryptococcal meningitis is one of the major complications affecting the central nervous system of patients suffering from AIDS. The results of treatment, when following current recommendation are still unsatisfactory. OBJECTIVE: This study aimed to evaluate the efficacy of a higher than recommended dose of oral fluconazole and itraconazole as consolidation therapy for cryptococcal meningitis in AIDS patients. DESIGN AND METHOD: HIV infected patients with primary cryptococcal meningitis, who had been treated initially with amphotericin B for 2 weeks were included in this study. They were randomized into two groups, to receive either fluconazole 600 mg daily or itraconazole 600 mg daily for 10 weeks. The response towards the two different treatments was clinically defined to be successful, if after 10 weeks of treatment no clinical symptoms and signs of meningitis remained and the cerebrospinal fluid (CSF) fungal culture was negative. RESULTS: The trial was performed from April 1999 to April 2000 at Srinagarind Hospital, Khon Kaen, Thailand. At the beginning of the trial 44 cases were selected, but only 35 patients proved to be suitable for the final evaluation of the study. Out of those, 19 cases were assigned to the fluconazole and 16 cases to the itraconazole group. Ten weeks after treatment, all patients clinically recovered completely. The CSF sterilization rate for the fluconazole group and for the itraconazole group were 100 and 94 per cent respectively. The Fisher's exact test showed no significant difference in the CSF sterilization rate between both groups (p = 0.26). CONCLUSION: The result of this study indicates that treatment with either 600 mg per day of fluconazole or itraconazole as consolidation treatment have the same efficacy for AIDS patients suffering from cryptococcal meningitis. The results of this study also suggest, comparing the result of this trial with the results of similar trials published somewhere else, that treatment with the higher doses may be superior to treatment regimens using lower doses, as can be judged from the clinical outcome and the results of the mycological cultures. PMID- 12757073 TI - Evaluation of attitude, risk behavior and expectations among Thai participants in Phase I/II HIV/AIDS vaccine trials. AB - The Understanding of volunteers in vaccine trials about their role as study participants and their voluntary commitment during the study are always one of the important concerns apart from evaluation of safety and efficacy of vaccine trials, especially in HIV prophylactic vaccine trials. The apprehension of indirectly risky behavior encouragement and deviated expectations among volunteers should be of concern. The current prospective cohort study aimed to assess and monitor the changes of risk behaviors, attitude and expectations among 164 volunteers from 2 studies of different prophylactic HIV vaccines, the Chiron HIV Thai E gp 120/MF59 +/- the Chiron HIV SF52 gp120 and Aventis Pasteur Live Recombinant ALVAC HIV (vCP1521) priming with VaxGen gp120B/E (AIDSVAX B/E) boosting. 113 males and 51 females with a mean age (+/- SD) of 28.82 +/- 7.97 years old were enrolled from October 1997 to December 1998 and February 2000 to April 2001. Education and risk reduction counseling were regularly performed at every visit and questionnaires about risk behaviors, knowledge, attitudes, social influences and expectations were asked at baseline, 4 months and 12 months. No change of potentially HIV transmission related risk behavior was observed during the studies. There was a statistically significant decrease of risk sexual practices from the beginning of the trials (42.2% vs 1%, p < 0.0001). While 35.2 per cent from 62.2 per cent of the volunteers at the beginning of the study continued sexual practice with an identified single sexual partner at the end of the study (p < 0.0001). All of the volunteers expressed the beneficial expectations as knowledge gain, social contribution, feelings of having gained merit and self-benefits from health check-ups. PMID- 12757074 TI - A randomized controlled trial of pubovaginal sling versus vaginal wall sling for stress urinary incontinence. AB - PURPOSE: To compare the results of Pubovaginal sling and Vaginal wall sling for the treatment of stress urinary incontinence in females. MATERIAL AND METHOD: Between February 2001 and December 2001, a randomized controlled trial was done to compare safety and efficacy of pubovaginal sling versus vaginal wall sling in the management of women with urinary incontinence. Fifteen women 42-68 years old (mean age 51.3 years) were treated with fascial sling (group A) and 11 women 45 60 years old (mean age 50.4 years) with vaginal wall sling (group B). Twenty-one patients had type II SUI and 5 patients had type III SUI (ISD); none had pre operative detrusor instability. Measures of outcome included efficacy based on SEAPI-QMN, post-operative presence of stress or urge incontinence, frequency of complications, operative time, post-operative pain, length of hospitalization, length of clean intermittent catheterization (CIC) time and mean global evaluation. RESULTS: All patients were followed for at least 3 months after surgery (median 7 months). A total of 20 and 6 women received spinal and general anesthesia, respectively. SEAPI-QMN decreased from a median of 6.3 to 0.8 for group A and from 6.1 to 0.9 for group B. No patient in either group had persistent stress incontinence. Urge incontinence was present in 2 of group A patients and 1 of group B patients. No serious post-operative complications were encountered in both groups. Post-operative pain and operative times for group B patients were significantly lower than for group A patients. Length of hospitalization, length of CIC time and mean global evaluation were not significantly different between the two groups. CONCLUSION: In the short-term, both pubovaginal sling and vaginal wall slings were effective in the treatment of women with SUI. However, the use of vaginal wall sling resulted in significantly shorter operative times and lower post-operative pain compared with pubovaginal sling. Therefore, the vaginal wall sling should be the prefered treatment for SUI. PMID- 12757075 TI - An epidemiological study on insomnia in an elderly Thai population. AB - The authors investigated the one-month prevalence and associations of insomnia in an elderly Thai population. A random sample of 40,111 individuals was selected from those of persons over 60 years of age by multiple stage sampling. The subjects were interviewed using a sleep questionnaire. Prevalence of insomnia of the population was 46.3 per cent. Depression and poor perceived health were factors strongly associated with insomnia. On the basis of these findings, the authors consider the prevalence of insomnia among the Thai elderly to be rather high. The implications of this study are of great importance for the design and development of preventive strategies and community-based interventions. PMID- 12757076 TI - Mastectomy without drain at pectoral area: a randomized controlled trial. AB - OBJECTIVES: Mastectomy is still one of the standard alternative procedures for the management of female breast cancer. Axillary node dissection is also performed to establish the accurate staging. After operation, the axilla must be drained because of lymphatic leakage. Whether the raw surface at the pectoral area should be drained or not is an interesting controversial point. The authors conducted a randomized controlled trial to compare outcomes after modified radical mastectomy (MRM) with and without drainage at the pectoral area. METHODS: Sixty patients who agreed to be treated with MRM and had given their consent were enrolled. Mastectomy was performed to remove the breast tissue proper by scalpel in order to minimize tissue injury. The axillary contents were removed by sharp instrument. After bleeding had stopped, patients were randomly allocated to one or other of 2 groups: group I (n = 30): only 1 drain was inserted at the axilla area; group II (n = 30): 2 conventional drains were inserted into the pectoral area and axilla area. The size of tube drain and negative suction pressure were constant in all cases. Volume of contents was recorded daily. Subcutaneous seroma or hematoma were carefully observed and confirmed by ultrasonography 3-5 days after operation. Overall drainage contents and complications were compared. RESULTS: The mean weight of breast tissue of group I was 632.1 g and group II 654.0 g (p = 0.81). Total drainage contents (median) from the two groups were 250 cm3 and 231 cm3 respectively (p = 0.796). Complications occurred in 1 case in group I and 2 cases in group II (p = 0.35). None of the above differences were statistically significant. CONCLUSION: Mastectomy by scalpel can be performed without drainage at the pectoral area. Overall complications in the conventional group and the group without drain did not differ significantly. PMID- 12757077 TI - Sonographic morphological pattern in the pre-operative prediction of ovarian masses. AB - The study was undertaken to evaluate the accuracy of sonographic morphological pattern in the detection of ovarian malignancy. A total of 123 patients with a suspicion of ovarian pathology, who were scheduled for elective surgery at the Department of Obstetrics and Gynecology, Faculty of Medicine Siriraj Hospital, Mahidol University were included in the study. All patients underwent sonographic examination prior to surgery by the same physician. The sonographic morphological pattern of each patient was compared to the histological diagnosis of the ovarian tumors. Of the 120 patients with an ovarian lesion, the sonographic morphological pattern of 10 had a sensitivity of 88.6 per cent and a specificity of 89.4 per cent in detection of malignant ovarian tumors. The positive predictive value, negative predictive value, and the accuracy rate were 77.5, 95.0, and 89.2 per cent, respectively. In the present study, a score of 9 would be the best discriminator between benign and malignant ovarian masses, giving a sensitivity of 97.1 per cent and specificity of 82.4 per cent. As stand alone, the present results confirm that ultrasonography is still a useful diagnostic tool in the differentiation of benign from malignant ovarian masses. PMID- 12757078 TI - Pythium corneal ulcer in Ramathibodi Hospital. AB - This is a retrospective study of corneal ulcers caused by Pythium insidiosum in Ramathibodi Hospital from 1988 to 1998. The clinical data was from the medical records of 10 patients, of which 8 had complete information. Mean age of the patients was 49.8 years old and seven were farmers. Initial visual acuity was mostly below finger counts. All manifested as fungal corneal ulcers, diagnosis of Pythium insidiosum was confirmed by histology. After failed medication, penetrating keratoplasty (PKP) was performed in order to remove the infected tissue. One patient had only anterior lamella keratectomy performed and was completely cured. Seven other patients had to have their eyes removed (evisceration or enucleation) to be cured. The study shows that Pythium corneal ulcer is rare but devastating. Patients with an agricultural occupation are most at risk. Antibiotics and antifungals could not control or cure this disease, so education for people at high risk is advocated. PMID- 12757079 TI - Body iron stores in Thai women of reproductive age. AB - Body iron stores is sensitively indicated to iron status. Since iron status strongly affects to iron absorption, body iron stores is a factor of estimating the dietary iron absorption method as proposed by Monsen. This study aimed to determine body iron stores in Thai women of reproductive age, which is one of the iron deficiency risk groups. The serum ferritin levels of 115 normal iron status (serum ferritin > or = 12 ng/ml) women aged between 18-45 years were included in the body iron stores calculation by Cook's method. The result showed that the mean body iron stores of the women was 292.78 mg. This finding was consistent with previous reports that the values were in the range of 200-400 mg in the women. The mean body iron stores were of 309 mg and nearly 300 mg in American and Australian women, respectively. The values were less than the recommended amount of 500 mg in adult women. Only 3 per cent of the Thai women in the present study met the recommended amount. So, as in American and Swedish women, less than 5 per cent of them reached the storage iron of 500 mg. PMID- 12757080 TI - Basic tear secretion measurement in pterygium. AB - PURPOSE: To evaluate the correlation between dry eye and pterygium. MATERIAL AND METHOD: A prospective study of basic tear secretion in 30 patients who had unilateral pterygium was performed. Schirmer's test with anesthesia was assessed in both eyes of patients with unilateral pterygium. RESULTS: Mean Schirmer's test value in the eye with pterygium was 11.6 +/- 0.4 mm and 12.4 + 0.4 mm, without pterygium. The Schirmer's test value was decreased significantly in the eye with pterygium. CONCLUSION: This study revealed that the Schirmer's test value with anesthesia was decreased significantly in the eye with unilateral pterygium when compared with a healthy eye. PMID- 12757081 TI - Uroflowmetry in normal Thai subjects. AB - BACKGROUND AND OBJECTIVES: Uroflowmetric parameters of urination may be influenced by many factors including age, sex, voiding position, technique used and also anatomical and physiological variations. A cross-sectional study was carried out to measure uroflowmetric parameters in normal Thai subjects and to compare these parameters among different ages and genders. Correlations between peak flow rate and other parameters were also studied. METHOD: One hundred and forty healthy Thai subjects were studied. They were classified into two groups. Group I comprised of 50 male and 50 female young adults aged 18-30 years. Group II comprised of 20 male and 20 female pre-elderly aged 50-60 years. A Dantec Urodyn 1,000 uroflowmeter was used. The residual urine measurement was performed using an ultrasonograph. RESULT: The techniques revealed the following uroflowmetric parameters. In the young adults, the mean with standard deviation of the peak flow rate was 31.2 +/- 9.0 ml/sec, mean flow rate 22.6 +/- 7.4 ml/sec, voiding time 24.7 +/- 10.6 sec, and voided volume 376.9 +/- 147.5 ml. In the pre-elderly group, the peak flow rate was 27.5 +/- 9.2 ml/sec, mean flow rate 19.1 +/- 6.2 ml/sec, voiding time 24.4 +/- 8.5 sec, and voided volume 310.3 +/- 107.8 ml. The peak flow and mean flow rates were significantly higher in the young adults (p < 0.05). The voided volume in the young was higher with similar voiding time. The peak flow and mean flow rates in females were significantly higher than the males (32.5 +/- 10.0 vs 27.8 +/- 8.0 ml/sec, p < 0.05 and 23.5 +/ 8.1 vs 19.8 +/- 5.8 ml/sec, p < 0.05 respectively). Voided volume and voiding time did not differ among both genders. The correlation between peak flow rate and voided volume was significant (r = 0.382, p < 0.01) indicating that the higher the voided volume the higher the peak flow rate. Residual urine was less than 50 ml in all subjects indicating that these subjects could void completely well. This study yielded normal uroflowmetric parameters in Thai young adult and pre-elderly subjects without urological symptoms. These parameters vary with age and gender, and are useful for the investigations of bladder function in a urological clinic. PMID- 12757082 TI - Isolated primary chylopericardium: a case report. AB - Isolated primary chylopericardium is a rare entity with an obscure etiology. The authors report a 10-week-old male infant presenting with tachypnea and enlarged cardiac silhouette. Echocardiography revealed a large pericardial effusion. A specific diagnosis of chylopericardium was made by pericardiocentesis and analysis of the fluid. Despite the pericardial tube drainage and medium-chain triglyceride diet, pericardial effusion reaccumulated. Ligation of the thoracic duct with the establishment of a pleuropericardial window was performed through a left thoracotomy. Follow-up echocardiograms have shown no reaccumulation of the pericardial fluid. PMID- 12757083 TI - Pseudoaneurysm following modified Blalock-Taussig shunt: a rare complication mimicking pulmonary disease. AB - The authors report a pseudoaneurysm in a 2-year-old boy presenting with fever, increasing cyanosis and right upper lung shadowing on a chest radiograph at six weeks following modified Blalock-Taussig shunt surgery. Echocardiography and a CT scan of the chest revealed a large pseudoaneurysm originating from the right subclavian artery at the proximal insertion of modified Blalock-Taussig shunt. The patient underwent aneurysmal resection, Blalock-Taussig shunt removal, right subclavian artery ligation and the creation of a central shunt between the ascending aorta and main pulmonary artery. Unfortunately, the patient died 3 hours after the operation. PMID- 12757084 TI - Balloon-expandable stenting, percutaneous coil embolization, and amplatzer septal occluder; treatments after complicated lateral tunnel Fontan: a case report. AB - The authors report a 7-year-old girl with univentricular heart physiology who developed prolonged pleural effusion due to discrete narrowing of the proximal right pulmonary artery, and progressive cyanosis which resulted from leakage of the atrial baffle, multiple veno-venous collaterals after the lateral tunnel Fontan operation. Percutaneous balloon-expandable stent implantation was used to correct the right pulmonary artery stenosis with an excellent result. Cyanosis was improved by coil embolization of the collaterals and occlusion of the baffle leakage with Amplatzer septal occluder. This is the first successful report in Thailand. PMID- 12757085 TI - Episodic ataxia type 2: an uncommon inherited CNS channelopathies. AB - The author reports the first Thai patient with a rare inherited ataxic disorder characterized by intermittent episodes of ataxia, headache and vertigo. The patient was well between attacks despite persistent nystagmus on examination. Magnetic resonance imaging of the brain revealed cerebellar atrophy. All symptoms were ameliorated by acetazolamide therapy. This clinical syndrome was previously described as acetazolamide-responsive episodic ataxia which was subsequently shown to be associated with mutations in a alpha1A-subunit of P/Q type voltage gated calcium channel gene, known as 'episodic ataxia type 2'. Clinical and molecular aspects of episodic ataxia type 2 were also reviewed. PMID- 12757086 TI - Congenital aortocaval fistula to the superior vena cava: a case report. AB - Various systemic arteriovenous fistulas have been described. The arteriovenous fistula arising from the ascending aorta and draining separately into the superior vena cava is very uncommon. The authors report a case of congenital aortocaval fistula to the superior vena cava in a 22 year-old woman in whom the fistula was closed successfully. PMID- 12757087 TI - Pleuropulmonary blastoma in a child presenting with spontaneous pneumothorax. AB - The authors described a 27-month-old boy with the diagnosis of pleuropulmonary blastoma who presented with spontaneous pneumothorax. The child was admitted to our hospital with the chief complaint of respiratory distress for 8 months. Initial chest X-ray revealed tension pneumothorax on the right side. After chest tube insertion to the right side, a repeated chest X-ray showed minimal pleural effusion and a mass-like lesion at the right lower lung field. Computed tomography (CT) of the chest showed a cavity with intramural mass confined in the right lower lung accompanied with hydropneumothorax. The surgery revealed a cystic and solid mass occupying the right pleural space medially displacing the right lower lung. Total removal of the mass was performed, the histopathologic findings revealed a mixed cystic and solid type of pleuropulmonary blastoma which was composed of primitive blastema with multidirectional differentiation. Combination chemotherapy consisting of vincristine, cyclophosphamide, doxorubicin and dactinomycin was administered two weeks after surgery. The child has been well for almost 6 months since the surgery, without any signs of metastasis or recurrence. PMID- 12757088 TI - Ethical consideration for the senior physician. PMID- 12757089 TI - Comparison of intraocular pressure profiles during cataract surgery by phacoemulsification and extracapsular cataract extraction. AB - BACKGROUND AND OBJECTIVE: In view of reports of fluctuations in intraocular pressure (IOP) during phacoemulsification, real-time IOP during small-incision, manual extracapsular cataract extraction (ECCE) and phacoemulsification was measured to determine the effect of anterior chamber maintainer infusion on intraoperative IOP dynamics. PATIENTS AND METHODS: A comparative study of IOP during cataract surgery was conducted by these two methods in 29 patients. An anterior chamber maintainer infused balanced salt solution continuously into the eye throughout ECCE. The anterior chamber maintainer was open in 7 cases throughout phacoemulsification and switched off during phacoemulsification in 7 cases. RESULTS: IOP measurements during small-incision ECCE were physiologic (range, 5 to 20 mm Hg) during most of the operation and coincided with the value of hydrostatic pressure established by the height of the anterior chamber maintainer infusion bottle. During phacoemulsification without concomitant anterior chamber maintainer, IOP was positive (> 2 to 3 mm Hg) as long as the phacoemulsification infusion line was activated (position 1). Moving to position 0 caused hypotony. Occlusion breaks occasionally caused wide fluctuations in IOP. Concomitant anterior chamber maintainer and phacoemulsification infusion did not prevent surges, but may account for the lower range of fluctuation. The anterior chamber maintainer also maintained a positive IOP at position 0 during phacoemulsification. CONCLUSIONS: An anterior chamber maintainer in conjunction with a self-sealing incision can maintain stable physiologic IOP during small incision ECCE. The IOP fluctuations during phacoemulsification, especially during occlusion breaks, can be attenuated by an anterior chamber maintainer. PMID- 12757091 TI - Evaluation of limbal conjunctival autograft and low-dose mitomycin C in the treatment of recurrent pterygium. AB - BACKGROUND AND OBJECTIVE: To assess the effectiveness of limbal conjunctival autograft and low-dose mitomycin C in the treatment of recurrent pterygium. PATIENTS AND METHODS: Thirty-four eyes with recurrent pterygium underwent surgical excision followed by the intraoperative application of low-dose (0.05%) mitomycin C and the use of limbal conjunctival autograft. All patients were observed for at least 18 months for any complications, including recurrences. RESULTS: Neither recurrences nor serious complications were recorded. Visual acuity improved significantly (P < .05) in eyes with pterygium extending 3 mm or more beyond the limbus. CONCLUSION: Simple excision and low-dose mitomycin C followed by limbal conjunctival autograft is a safe and effective way of treating recurrent pterygium. PMID- 12757090 TI - Intravitreal dexamethasone effectively reduces postoperative inflammation after vitreoretinal surgery. AB - BACKGROUND AND OBJECTIVE: Corticosteroids remain the mainstay for control of ocular inflammation after vitreous surgery. A controlled, randomized, prospective study was performed to evaluate the effectiveness of a single intravitreal injection of dexamethasone phosphate on postoperative inflammation after simple vitreous surgery in patients with proliferative diabetic retinopathy and macular pucker. PATIENTS AND METHODS: Aqueous cell flare intensity was measured preoperatively and on days 1, 10, and 90 in 56 consecutive patients who underwent vitreous surgery for proliferative diabetic retinopathy and macular pucker. Subjects were consecutively randomized to two groups: 400 microg of intravitreal dexamethasone (treatment group) or no dexamethasone (control group) RESULTS: Before surgery, cell and flare intensity was similar in both groups. Flare intensity was significandy lower at 10, 30, and 90 days in the proliferative diabetic retinopathy treatment group (P < .05). CONCLUSION: Intravitreal dexamethasone significantly alleviates postoperative inflammation after vitreous surgery and is a useful adjunct. PMID- 12757092 TI - Opacification of two hydrophilic acrylic intraocular lenses 3 months after implantation. AB - BACKGROUND AND OBJECTIVE: To report clinical, pathologic, histochemical, ultrastructural, and spectrographic analyses of explanted hydrophilic acrylic intraocular lenses (IOLs) obtained from two patients who had early visual disturbances caused by postoperative opacification of the lens optic. MATERIALS AND METHODS: Two hydrophilic IOLs were explanted from patients with decreased visual acuity. The patients became symptomatic 3 months after uneventful phacoemulsification and lens implantation. IOL optic opacification was associated with a fine granularity within the substance of the lens optic. The IOLs were examined by gross and light microscopy. Full-thickness cut sections of the optics were stained with 1% alizarin red (a special stain for calcium). Some sections were submitted for scanning electron microscopy and energy dispersive x-ray spectroscopy. RESULTS: Microscopic analyses revealed the presence of multiple fine, granular deposits of variable sizes within the optics of the lenses distributed in a line parallel to the anterior and posterior curvatures of the optic, with a clear zone just beneath the optics' surface. Extension of the opacification into the haptics of the IOLs could also be observed. The deposits stained positive with alizarin red. Energy dispersive x-ray spectroscopy of the internal substance of the IOLs also demonstrated the presence of calcium within the deposits. CONCLUSIONS: This is the first clinicopathologic report of optic and haptic opacification occurring with this hydrophilic acrylic IOL model only 3 months postoperatively. Further studies on other similar cases with this lens should be done to determine the incidence and possible mechanisms of this phenomenon. PMID- 12757093 TI - Nd:YAG laser photocystotomy of a free-floating pigmented anterior vitreous cyst. AB - An 8-year-old girl presented with a free-floating pigmented cyst in the anterior vitreous causing a large, visually disturbing floater. A simple laser photocystotomy with a single-frequency Nd:YAG laser resulted in a reduction in the size of the cyst but persistence of the floater. An attempt at further laser fragmentation of the cyst resulted in iatrogenic cataract formation. PMID- 12757094 TI - Rhegmatogenous retinal detachment with Schwartz's syndrome following Nd:YAG laser peripheral iridectomy in the management of pigmentary glaucoma. AB - A case of rhegmatogenous retinal detachment complicated by Schwartz's syndrome following Nd:YAG laser peripheral iridectomy in the management of pigmentary glaucoma is described. A 36-year-old man underwent Nd:YAG laser peripheral iridectomy for pigmentary glaucoma. Three days later, he had a rhegmatogenous retinal detachment. The break remained open following scleral buckling with persistent subretinal fluid, "iridocyclitis," and elevated intraocular pressure. Two months later, the patient underwent revision of the scleral buckle. Following closure of the retinal break, the retina flattened, and within 2 weeks the "iridocyditis" resolved and the intraocular pressure decreased to approximately 20 mm Hg. Rhegmatogenous retinal detachment may occur following laser peripheral iridectomy, and suggests the necessity for a prospective dinical trial to evaluate the role of laser peripheral iridectomy in the management of pigmentary glaucoma. PMID- 12757095 TI - Delayed suprachoroidal hemorrhage following viscocanalostomy. AB - Delayed suprachoroidal hemorrhage is a rare complication of conventional glaucoma surgery. Viscocanalostomy is one of the new surgical techniques used in glaucoma surgery that may theoretically protect against suprachoroidal effusion and hemorrhage due to the nonpenetrating nature of the procedure. Delayed suprachoroidal hemorrhage developed in a 92-year-old white woman following viscocanalostomy. This case demonstrates that the risk of suprachoroidal hemorrhage may not be completely eliminated after a nonpenetrating glaucoma procedure such as viscocanalostomy. PMID- 12757096 TI - Intraretinal hemorrhage associated with leishmaniasis. AB - A 39-year-old man presented with sudden loss of visual acuity caused by two retinal hemorrhages with no choroidal neovascularization (confirmed by fluorescein angiography). The patient was hospitalized for malaise, progressive pancytopenia, hepatosplenomegaly, progressive anemia, and perianal inflammation. Positive serologies were obtained for toxoplasmosis (IgG) and Leishmania (1/160). A diagnosis of visceral leishmaniasis was made, and the patient was treated with pentavalent antimonials. Two months later, best-corrected visual acuity was 20/25, with no residual scotoma. Indirect ophthalmoscopy showed complete resolution of the hemorrhages. This patient was an otherwise healthy immunocompetent adult who presented frank visceral leishmaniasis and retinal hemorrhages as the only ocular or systemic hemorrhagic findings, with spontaneous resolution after improvement of platelet levels. This rare cause of macular hemorrhage should be considered in areas where Leishmania is endemic. PMID- 12757097 TI - Central retinal vein occlusion immediately following cardiac surgery. AB - The relationship between cardiac surgery and central retinal vein occlusion (CRVO) has been previously documented; however, in all of these cases the occlusion occurred months after the cardiac surgery. This is the first report of a patient awakening from cardiac surgery with ophthalmoscopically documented CRVO. This diagnostic case report describes a patient who developed CRVO immediately following extensive cardiac surgery including aortic valve, aortic root, and proximal aortic arch replacements, as well as coronary artery bypass grafts. Ophthalmologists and cardiac surgeons alike should be aware that CRVO is a potential complication of cardiac surgery, especially when cardiopulmonary bypass is necessary. PMID- 12757098 TI - Retinal pigment epithelial tear involving the fovea with preserved visual function. AB - Tears of the retinal pigment epithelium are known to occur either spontaneously or after laser photocoagulation in eyes with retinal pigment epithelium detachment. A 65-year-old man with preexisting retinal pigment epithelium detachment developed a retinal pigment epithelium tear after dye laser retinal photocoagulation. The tear gradually expanded to involve the fovea, but his best corrected visual acuity remained 0.7 in the left eye during 20 months. Optical coherence tomography showed a defect of the retinal pigment epithelium with absence of regeneration. Scanning laser ophthalmoscopy revealed his fixation approached intact retinal pigment epithelium, but was still beneath the fovea. This case may indicate that the retinal pigment epithelium directly beneath the central macula is not essential for maintenance of the overlying foveal function under some conditions. PMID- 12757099 TI - A newborn with lipemia rednalis. AB - Lipemia retinalis is a rare ocular manifestation of certain types of hyperlipidemia. A case of a newborn with lipemia retinalis evaluated by laboratory and dinical findings is described. A creamy white appearance of all retinal blood vessels was demonstrated by color fundus photographs. The patient had elevated levels of triglycerides (29,000 mg/dL) and cholesterol (1,470 mg/dL). Lipid electrophoresis indicated hyperprebetalipoproteinemia. Type IV primary hyperlipoproteinemia was diagnosed. PMID- 12757100 TI - Intraocular intrusion of a scleral sponge implant. AB - A patient with intrusion of a scleral sponge implant after retinal reattachment surgery is described. Retinal incarceration occurred during primary retinal reattachment surgery and the patient underwent additional cryopexy and radial sponge implant. However, the retina remained detached and pars plana vitrectomy was performed with silicone oil endotamponade to reattach the retina. The interval between placement of the sderal buckle and development of intrusion was 3 months. The buckle was sutured to the sclera with mattress sutures. During the postoperative follow-up period, the intraocular pressure was elevated and controlled with topical antiglaucomatous medication. The encircling band was cut but left in place to avoid perforation, and the retina remained attached. Intrusion of a buckle after retinal reattachment surgery is a rare complication that usually does not require surgical intervention. Cutting the silicone band without removing it may be a treatment option. PMID- 12757102 TI - Surgical management of unilateral elevator deficiency associated with horizontal deviation using a modified Knapp's procedure. AB - Both horizontal recti are transpositioned in unilateral elevator deficiency, leaving no alternative to correct any significant ipsilateral horizontal deviation. An effective method to correct vertical and horizontal deviations simultaneously is reported. A modified Knapp's procedure was performed by transposition of the superior half of equally divided (up to 15 mm) medial and lateral recti for vertical deviation and the inferior half after suitable recession or resection for horizontal deviation. This procedure was performed in ten patients who had type II unilateral elevator deficiency with horizontal deviation ranging from 18 to 45 prism diopters (PD). We were able to correct 20 PD of horizontal deviation and 25 PD of vertical deviation with a modified Knapp's procedure without any significant adverse effects after a follow-up of 21 months. A modified Knapp's procedure is recommended for unilateral elevator deficiency associated with horizontal deviation. PMID- 12757103 TI - Trypan blue staining of the anterior capsule under an air bubble with a modified cannula. AB - To attain good visibility of the anterior capsule in the advanced or white cataract, trypan blue 0.1% is used to stain the anterior capsule. The dye is usually injected under an air bubble. However, it is difficult to inject the dye properly due to capillary forces. An ordinary anterior chamber cannula was modified and its coverage area increased to facilitate the staining of the anterior capsule under an air bubble. The anterior capsule was stained properly by using the modified cannula in all of the cases. PMID- 12757101 TI - Temporary use of a customized, glued-on hard contact lens before penetrating keratoplasty for descemetocele or corneal perforation. AB - Descemetocele or corneal perforation makes it difficult to perform penetrating keratoplasty (PKP). To circumvent this difficulty, a polymethylmethacrylate hard contact lens with a diameter of 4.0 mm was customized and applied to the cornea with tissue adhesive prior to PKP in three patients, one with corneal perforation and two with descemetoceles. The results showed that this modified method facilitated trephination during PKP without complications in all three patients, suggesting that it may be applicable to other similar clinical situations. PMID- 12757104 TI - Graphic representation of data resulting from measurement comparison trials in cataract and refractive surgery. AB - BACKGROUND AND OBJECTIVES: The evaluation of new diagnostic measurement devices allows intraindividual comparison with an established standard method. However, reports in journal articles often omit the adequate incorporation of the intraindividual design into the graphic representation. PATIENTS AND METHODS: This article illustrates the drawbacks and the possible erroneous conclusions caused by this misleading practice in terms of recent method comparison data resulting from axial length measurement in 220 consecutive patients by both applanation ultrasound and partial coherence interferometry. RESULTS: Graphic representation of such method comparison data should be based on boxplots for intraindividual differences or on Bland-Altman plots. Otherwise, severe deviations between the measurement devices could be erroneously ignored and false positive conclusions on the concordance of the instruments could result. CONCLUSION: Graphic representation of method comparison data should sensitively incorporate the underlying study design for intraindividual comparison. PMID- 12757105 TI - Scanning laser Doppler flowmetry of nonperfused regions of the optic nerve head in patients with glaucoma. AB - BACKGROUND AND OBJECTIVE: To examine the reliability of scanning laser Doppler flowmetry in measuring capillary flow in regions of the optic nerve head, which are manifested differently by confocal tomographic angiography. PATIENTS AND METHODS: Capillary blood flow of the optic nerve head was measured in 16 eyes of 13 patients who had glaucoma with moderate to severe visual field defects. Blood flow measurements were taken by the scanning laser Doppler flowmeter and analyzed using the automatic full-field perfusion image analyzer (AFFPIA). Flow values were compared between perfused and nonperfused halves of the optic nerve head. Vascular anatomy of the optic nerve head was evaluated by confocal laser tomography with indocyanine green injection, and visual field tests by a standard visual field analyzer. RESULTS: Analysis of laser capillary flow parameters by the AFFPIA disclosed a mean lower flow value in nonperfused compared with perfused discs (426 +/- 310 vs 547 +/- 306; P = .006). There was also a positive correlation between visual field defect and capillary angiography of the optic nerve head. CONCLUSIONS: Flow measurements of the optic nerve head by scanning laser Doppler flowmetry using the AFFPIA revealed a mean lower flow value in nonperfused discs of patients who had glaucoma with severe visual field defect. Capillary angiography demonstrated by Heidelberg retina angiography has a good correlation with the pattern of visual field defect. PMID- 12757106 TI - Evaluation of hydroxychloroquine retinopathy with multifocal electroretinography. AB - BACKGROUND AND OBJECTIVE: To describe the changes revealed by multifocal electroretinography (ERG) in patients taking hydroxychloroquine. PATIENTS AND METHODS: Six patients being treated for various inflammatory conditions with hydroxychloroquine for periods ranging from 8 months to 7 years were consecutively evaluated. Each examination included measurement of Snellen visual acuities, Amsler grid assessment, and automated visual field testing. In some cases, funduscopic examinations were complimented by photography and fluorescein angiography. Multifocal ERG was performed for all patients. RESULTS: Three patients (six eyes) were found to have distinctive abnormalities on multifocal ERG consisting of pericentral depression of ERG signals. The abnormalities on multifocal ERG corresponded with the patients' subjective descriptions and the visual field depiction of their pericentral scotomas. All affected patients had been taking hydroxychloroquine for at least 7 years. One patient with generalized depression on multifocal ERG had possible hydroxychloroquine retinopathy. Two patients (three eyes) had relatively normal results on multifocal ERG. CONCLUSION: Multifocal ERG objectively demonstrates depression of signals in the perifoveal region in visually symptomatic patients with long-term hydroxychloroquine use. Even patients with normal visual acuity and no fundus abnormalities can have abnormal results. Although we have not yet identified patients with abnormalities on multifocal ERG before the onset of symptoms, multifocal ERG may be useful in monitoring patients at risk and may provide an earlier opportunity to identify maculopathy. PMID- 12757107 TI - High-frequency ultrasonographic imaging in suprachoroidal hemorrhage after filtering surgery. AB - A 64-year-old woman with glaucoma had suprachoroidal hemorrhage approximately 6 hours after trabeculectomy. High-frequency ultrasonography revealed high reflectivity in the inner space of a kissing choroidal detachment, which facilitated the differential diagnosis of suprachoroidal hemorrhage resulting from serous choroidal detachment. The ciliary process and iris were anteriorly displaced due to the ciliary detachment and forward pressure of the anterior vitreous. On the basis of the diagnosis, transscleral choroidal drainage was performed without delay and the patient's vision was preserved. After surgery, suprachoroidal hemorrhage disappeared clinically, but high-frequency ultrasonography detected persistence of the choroidal detachment in the peripheral area. PMID- 12757108 TI - Specialized nutrition support: the pediatric perspective. AB - Nutrition support may be more important in children than in adults. The first reports of the use of parenteral nutrition and of parentaneous endoscopic gastrostomies were in children. A number of unresolved questions in this area remain and will be answered by a coordinated effort of the nutrition community. PMID- 12757109 TI - Clinical aspects of essential fatty acid metabolism: Jonathan Rhoads Lecture. AB - The clinical implications of the metabolism of the 2 essential fatty acids, linoleic and alpha-linolenic acid, are most clearly related to the membrane phospholipid concentrations of their elongation and desaturation products, arachidonic, eicosapentaenoic, and docosahexaenoic acid. Levels of these very long chain polyunsaturated fatty acids can be altered by diet, prematurity, and disease which can affect growth (nutritional repletion) and the intensity and character of systemic inflammation as well as cognitive and visual function in infants. PMID- 12757110 TI - Inhibition of activator protein-1 transcription factor activation by omega-3 fatty acid modulation of mitogen-activated protein kinase signaling kinases. AB - BACKGROUND: Lipopolysaccharide (LPS)-stimulated macrophages (Mphi) produce excess tumor necrosis factor (TNF), and the direct inhibition of IkappaB phosphorylation and its subsequent separation from the nuclear factor kappaB (NFkappaB)-IkappaB complex has been experimentally supported as a mechanism for omega-3 fatty acid (FA) inhibition of this TNF response. However, TNF production is a "late" event in the LPS-induced Mpsi inflammatory cascade, and in addition to NFkappaB associated pathways, a separate transcription factor, activator protein-1 (AP-1) is an important pathway for Mpsi proinflammatory cytokine production. The mitogen activated protein kinase (MAPK) cascade regulates both NFkappaB-IkappaB--and AP-1 associated gene transcription through several cross-amplifying phosphorylation kinases, specifically p44/42 [ie, extracellular signal-regulated kinase (ERK) 1/2], p38, and c//jun N-terminal kinase (JNK)/stress-activated protein kinase (SAPK). The activation of these kinases occurs in the proximal MAPK cascade and activation modulates AP-1 activation. In this set of experiments, it was hypothesized that inhibition of MAPK signaling phosphorylation kinases by omega-3 fatty acids in a model of LPS-stimulated Mphi(s) would alter the activation of the proinflammatory cytokine transcription factor AP-1. METHODS: RAW 264.7 cells were pretreated with a sterile, commercially available, pharmaceutical grade omega-3 FA emulsion, equivalent grade omega-6 FA emulsion, or Dulbecco's modified eagles medium (media alone) for 4 hours. Cells were washed twice and exposed to LPS for 15 minutes. Total cell lysates were collected, and both total and phosphorylated portions of the p44/42, p38, and JNK/SAPK proteins were determined by Western blotting. AP-1 nuclear translocation was determined by electromobility shift assay. RESULTS: Phosphorylation of p44/42 and JNK/SAPK proteins of the MAPK pathways in LPS-stimulated Mpsi(s) was significantly reduced by omega-3 FA treatment compared with Mphi treated with omega-6 FA or media alone. In contrast, phosphorylation of p38 was not inhibited in the presence of omega-3 or (omega-6 FA treatment compared with media alone. Omega-3 FA pretreatment inhibited AP-1 activation. CONCLUSIONS: omega-3 FA inhibited p44/42 and JNK/SAPK phosphorylation; however, p38 remained unchanged. Phosphorylation of p44/42 and JNK/SAPK are the immediate prior steps in AP-1 activation. Attenuated AP-1 activation and subsequent attenuated gene-level proinflammatory cytokine elaboration is anticipated after inhibition of these MAPK intermediates and is confirmed by the reduction in AP-1 activity. These results provide further evidence for the transcriptional level regulation in the elaboration of proinflammatory cytokines by omega-3 FA in this Mphi model. PMID- 12757111 TI - Oral glutamine and the healing of colonic anastomoses in rats. AB - BACKGROUND: Recent evidence has suggested that glutamine is one of the primary energy sources of the colon. The aim of this study was to evaluate the effects of oral glutamine supplementation on the healing of colonic anastomoses in rats. METHODS: Forty-eight adult male Wistar rats, weighing 174.41 +/- 37.39 g, were housed in individual cages. All rats had free access to water and standard rat chow. The rats were randomized to receive daily, for 7 days before the operation and during the postoperative period, 10% L-glutamine (GLN group) or 10% glycine (GLY group) in isonitrogenous and isovolumetric solutions (1.5 g/kg per day), through an orogastric tube. On the eighth day, rats were anesthetized and subjected to 2 colonic transections, one 6 cm distal from the ileocecal valve and another 5 cm distal from the first transection. Bowel continuity was restored by 2 end-to-end, single layer, everted, anastomoses with 8 interrupted sutures (6-0 nylon). After the operation, the rats were kept in individual cages and had free access to water and rat chow. One-half of the rats in each group were killed either on postoperative day 3 or 8, and the 2 colonic anastomoses of each animal were resected and stored in 0.9% saline and 10% formalin for tensile strength and histologic (hematoxylineosin and collagen densitometry) studies, respectively. Student's t-test and Kruskal Wallis tests were used for statistical analysis. RESULTS: Total rupture strength was significantly higher in the GLN group (GLN: 0.068 +/- 0.045 kgf versus GLY: 0.042 +/- 0.027 kgf, p = .04). The mean monocytes infiltrate was significantly smaller in the GLN group (p = .04). The collagen densitometry analysis demonstrated greater percent area of type I (mature) in the GLN group compared with GLY (58.65 +/- 11.70% versus 41.79 +/- 10.54%, p = .0000), respectively. Subgroup analyses according to the day of rat death were still significant: GLN 3: 54.22 +/- 10.02% versus GLY 3: 41.92 +/- 13.31% (p = .04) and GLN 8: 62.63 +/- 12.13% versus GLY 8: 41.67 +/- 7.69% (p = .0004). Type III collagen (immature) percent area was significantly smaller in the GLN group's colonic anastomoses (GLN: p = .0000; GLN 3: p = .04 and GLN 8: p = .0003, respectively). CONCLUSIONS: Perioperative oral glutamine supplementation increases total rupture strength and improves the percent area of mature collagen at the anastomoses sites on postoperative days 3 and 8. PMID- 12757112 TI - Glutamine infusion during ischemia is detrimental in a murine gut ischemia/reperfusion model. AB - BACKGROUND: Gut ischemia/reperfusion (I/R) frequently occurs in clinical settings as a result of disproportionate splanchnic hypoperfusion during shock. Glutamine (GLN) supplementation of total parenteral nutrition (TPN) before gut I/R improves survival after gut I/R compared with standard TPN. However, it is unknown whether GLN treatment after the occurrence of the insult is beneficial or not. The aims of this study were to examine effects of GLN infusion during gut ischemia on survival, myeloid cell (neutrophils + monocytes) activation, and vascular permeability in organs. METHODS: Male Institute of Cancer Research (ICR) mice were randomized to control and GLN groups. After IV cannulation, mice underwent 90 (experiments 1 and 2) or 60 (experiment 3) minutes of gut I/R. Control mice received normal saline infusion at 1 mL/h for 60 minutes during ischemia, whereas the GLN group was given 3% GLN solution. In experiment 1, survival rates were monitored for 72 hours (n = 25). In experiment 2, peripheral blood was obtained at 2 or 4 hours after reperfusion (n = 17). Reactive oxygen intermediate (ROI) production by myeloid cells was determined by flow cytometry using dihydrorhodamine 123 with or without phorbol myristate acetate stimulation. Expression of CD11a and CD11b on myeloid cells was also measured. Myeloperoxidase (MPO) activity in the lung was evaluated. In experiment 3, vascular permeability in organs was measured using Evans blue at 2 or 4 hours. RESULTS: In experiment 1, survival time in the GLN group was significantly reduced compared with the control group (p = .02, log-rank test). The survival rates were 92% (12/13) and 42% (5/12) for the control and GLN groups at 12 hours (p = .01) and 38% (5/13) and 0% (0/12) at 48 hours (p = .02), respectively. In experiment 2, ROI production was significantly higher in the GLN group than in the control group after PMA stimulation both at 2 and 4 hours. CD11b expression was significantly higher in the GLN group than in the control group at 4 hours. There was no difference in pulmonary MPO activity at either time point. In experiment 3, GLN infusion significantly increased hepatic vascular permeability compared with saline infusion at 4 hours. CONCLUSIONS: GLN infusion during ischemia is detrimental for survival after gut I/R. A possible mechanism is excessive priming of myeloid cells caused by GLN infusion. Timing of GLN administration is critical for outcome after gut ischemic insult. PMID- 12757113 TI - Brief refeeding rapidly reverses dietary restriction-induced nuclear factor kappaB downregulation in peritoneal resident cells. AB - BACKGROUND: Malnutrition impairs host immunity, resulting in high mortality and morbidity, secondary to infections. Nuclear factor kappaB (NFkappaB) plays a critical role in host defense, but how quickly refeeding normalizes the impaired NFkappaB activity in peritoneal resident cells (PRCs) is unknown. Our aim was to investigate the effects of 1-day ad libitum refeeding on severe diet restriction induced NFkappaB activity in PRCs. METHODS: Mice received chow, 146 g/kg per day (ad libitum) or 36.5 g/kg per day (severe diet restriction), for 7 days. One-half the mice in the diet-restricted group were then fed ad libitum for 1 day (refeeding). PRCs were harvested by peritoneal lavage. After incubation with tumor necrosis factor-alpha (TNF-alpha), nuclear translocation of NFkappaB in PRCs was investigated using laser scanning cytometry. RESULTS: The main subpopulation of PRCs was macrophages in all groups. Mean fluorescence intensity over the nuclear area at 0 or 100 ng/mL of TNF-alpha was 16 +/- 2 or 31 +/- 8* in the ad libitum, 20 +/- 4 or 19 +/- 3 in the severe diet-restricted, and 20 +/- 4 or 30 +/- 5* in the refeeding group, respectively (*p < .05 versus 0 ng/mL of TNF alpha in each group versus 100 ng/mL of TNF-alpha in diet-restricted group). Cytoplasmic accumulation of NFkappaB was significantly increased after TNF-alpha stimulation in the refed group but not in the ad libitum group. CONCLUSIONS: The blunted NFkappaB activity in PRCs, after exposure to inflammatory stimuli, was restored after 1 day of refeeding, with increased accumulation of NFkappaB in the cytoplasm. Even brief nutritional replenishment in malnutrition may improve host defense by restoring NFkappaB activity and thereby improving macrophage functions. PMID- 12757114 TI - 2003 Harry M. Vars Research Award. Keratinocyte growth factor improves epithelial function after massive small bowel resection. AB - BACKGROUND: Massive small bowel resection with subsequent short bowel syndrome (SBS) leads to the acute loss of epithelial cell (EC) absorptive function. Keratinocyte growth factor (KGF) has been shown to improve EC growth, although little is known about KGF activity on EC function after SBS. We hypothesized that KGF would improve epithelial function in a mouse SBS model. METHODS: Adult C57BL/6J mice were randomized to a 55% mid-small bowel resection (SBS), SBS with KGF administration (SBSKGF), or sham-operated (Control) group, and were killed at 7 days. Ussing chambers were used to study epithelial function. Short circuit current (Isc) was monitored. EC absorption was studied by measuring (1) glucose [3-O-methyl-D-[1-3H]glucose (3-OMG)] absorption; (2) sodium coupled amino acid (alanine) absorption; and (3) changes in Isc by using the absorptive agent D glucose (stimulated Na+ absorption). Epithelial barrier function was measured with transepithelial resistance (TER) and transmural passage of 3H-mannitol (Papp). ECs were separated along the crypt-villus axis with laser capture microdissection. Epithelial KGF receptor (KGFR) mRNA expression was studied using real time reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: KGF administration increased the basic ion transport activity and net transepithelial absorption of 3-OMG and sodium-coupled alanine absorption. SBS significantly decreased epithelial ion transport, including the Na+ absorption stimulated by D glucose and L-alanine. KGF administration partially improves Na+ absorption. KGF had no apparent effect on the TER and 3H-mannitol permeability in this study. KGF upregulated EC KGFR mRNA expression, predominately in the crypt and lower portion of the villus. CONCLUSIONS: KGF administration improves epithelial absorptive function and stimulates intestinal proliferation after SBS. This suggests that KGF improves intestinal adaptation after SBS and may have clinical applicability. PMID- 12757115 TI - Safety and metabolic tolerance of a concentrated long-chain triglyceride lipid emulsion in critically ill septic and trauma patients. AB - BACKGROUND: A concentrated fat emulsion (Intralipid 30%) with a phospholipid/triglyceride ratio of 0.04 was tested for clinical tolerance and metabolic effects in the short-term parenteral nutrition of septic and trauma critically ill patients and compared with Intralipid 20% (phospholipid/triglyceride ratio of 0.06). METHODS: This was a prospective, randomized, multicenter study in the intensive care units in 10 university hospitals, including 90 adult patients in 2 groups: 55 septic and 35 trauma patients. Patients in each group were randomly divided into 2 subgroups according to the fat emulsions administered (1.4 g/kg per day) as part of the calories for at least 6 days of continuous total parenteral nutrition (TPN). One subgroup was treated with 30% long-chain triglycerides (phospholipid/ triglyceride ratio: 0.04) and the other with 20% long-chain triglycerides (phospholipid/triglyceride ratio: 0.06). The parenteral nutrition formula was isocaloric and isonitrogenous with 0.25 g of nitrogen/kg per day and 40% of the nonprotein calories as fat. Clinical tolerance was assessed during the study. At baseline and after 3 and 6 days of TPN, the following biochemical parameters were measured: prealbumin, retinol-binding protein, serum albumin, hematologic, hepatic and renal function variables, triglycerides, phospholipids, total and free cholesterol, nonesterified cholesterol, nonesterified fatty acids, and lipoproteins. RESULTS: At baseline, no differences in age, gender, severity of the condition [Acute Physiology and Chronic Health Evaluation (APACHE II) score], or clinical chemistry were found between the subgroups. The levels of plasma proteins studied and the renal, hematologic, or hepatic function variables did not vary during the study period. Total cholesterol increased significantly, owing to esterified cholesterol, with 20% long-chain triglyceride in septic patients (baseline: 2.1 +/- 0.8 mmol/L, day 6: 2.8 +/- 0.6 mmol/L, p = .026). In septic patients receiving 20% long-chain triglycerides, plasma triglycerides had a similar behavior (baseline: 1.4 +/- 0.6 mmol/L, day 3: 2.2 +/- 0.8 mmol/L, p < .05). The very-low-density lipoprotein content of cholesterol, triglycerides, and phospholipids showed a tendency to decrease in septic patients treated with 30% long-chain triglycerides (NS). None of the emulsions induced the synthesis of lipoprotein X. CONCLUSIONS: Our results indicate that while both fat emulsions used in the TPN of critically ill patients are clinically safe, the 30% long chain triglyceride fat emulsion with a phospholipid/triglyceride ratio of 0.04 causes fewer lipid metabolic disturbances. PMID- 12757116 TI - Pump-assisted versus gravity-controlled enteral nutrition in long-term percutaneous endoscopic gastrostomy patients: a prospective controlled trial. AB - BACKGROUND: Vomiting, aspiration, flatulence, and diarrhea are well-known negative side effects of enteral nutrition through percutaneous endoscopic gastrostomy (PEG). However, it is not yet clarified if pump-assisted (PA) or gravity-controlled (GC) application is the more comfortable and safe choice for long-term nutrition through PEG. METHODS: This was a prospective, randomized, crossover study. Fifty long-term PEG patients were fed by PA nutrition (G1) and 50 patients were fed by GC nutrition (G2). Six weeks of observation (O1) was followed by a switch of method of nutritional application in both groups and an additional 6 weeks of observation (O2). Daily determination of comfort and safety was done with a standardized questionnaire. Evaluation of blood glucose levels on days 1, 21, and 42 during O1 and O2. RESULTS: The patients in both groups had the same medical conditions and were of the same age and sex. Far less flatulence (p < .0006) and epigastric fullness (p < .0003) was discovered in G1 during O1. Also, significantly less regurgitation (p < .0002) and vomiting of feeding diet (p < .0001) in G1 versus G2 could be observed. The rate of diarrhea (p < .0003) in G2 was higher than in G1. The daily profile of blood glucose was significantly better (p < .0008) in G1 than in G2. After the nutritional application was changed in O2, the PA group (G2) again showed a significantly better rate of flatulence, epigastric fullness, regurgitation, vomiting, diarrhea, and daily profile of blood glucose. Ninety-six percent of the patients in G2 preferred further nutrition by PA after finishing this study. All patients in G1 continued their accustomed nutrition by PA. CONCLUSION: Nutrition through PA showed not only a higher comfort rate but also increased safety, which was expressed through a low rate of regurgitation and vomiting. PA presented better glucose metabolization manifested in improved blood glucose levels. As a result of this prospective study, PA is preferable to GC and preferred by patients with long term PEG nutrition. PMID- 12757117 TI - New Food and Drug Administration requirements for inclusion of vitamin K in adult parenteral multivitamins. AB - The newly amended requirements by the Food and Drug Administration (FDA) for an effective adult parenteral multivitamin drug product increase the amounts of vitamins B1, B6, C, and folic acid currently in the product to better meet estimated needs, and specifies the inclusion of 150 microg of vitamin K. Infuvite Adult Multiple Vitamins for Infusion is the first adult parenteral multivitamin product to meet the revised FDA requirements. The inclusion of vitamin K in adult parenteral multivitamin products is intended to afford patients a consistent daily supply of vitamin K approximating usual levels of intake. This is a change to clinical practice in the United States, where vitamin K has not been included in adult parenteral vitamin preparations, and physicians prescribe vitamin K separately. The reformulation of adult parenteral multivitamins required by the FDA raises questions about the potential impact that inclusion of vitamin K will have on patient management. One clinical practice change is that patients on parenteral nutrition receiving Infuvite Adult should no longer need weekly subcutaneous (SC) or intramuscular (IM) vitamin K injections. In addition, the consistent and modest level of vitamin K provided by the reformulated adult parenteral multivitamins may make it easier for physicians to maintain the desired level of hypoprothrombinemia (low levels of prothrombin) in those patients also on anticoagulant therapy with warfarin. However, for physicians accustomed to administering vitamin K separately, it is important to highlight that it may be more difficult to titrate anticoagulant therapy, especially among patients receiving dual feeding or vitamin K from another source. If marketed globally, these clinical practice issues may be more pronounced outside of the United States, where dual feeding is common, and patients may receive vitamin K from other sources. PMID- 12757118 TI - Jonathan Roads Symposium Papers. History of parenteral nutrition. AB - A survey is given of the development of parenteral nutrition with the beginning of William Harvey's fantastic discovery of the circulation to today's discussion of what is an optimal regime of parenteral nutrition. The important and different steps of development during the 17th and 19th centuries are discussed. The modern steps during the last century leading to the concept we have today of parenteral nutrition is mentioned, with reference to all pioneers all over the world. Glucose, protein hydrolysates and crystalline amino acids, development of safe fat emulsions, and the current concepts of parenteral nutrition and future considerations are discussed. PMID- 12757120 TI - Dr. Robert T. Kennedy. PMID- 12757119 TI - What will the next healthcare system look like? PMID- 12757121 TI - Analysis of bipyridylium herbicides by capillary zone electrophoresis. AB - The separation of paraquat, diquat, and difenzoquat bipyridilium herbicides and ethyl viologen as internal standard is shown. The separation has been carried out using a fused silica capillary with a high-sensitivity detection cell for a capillary electrophoresis system and with a diode array detector. The experiments have been performed using phosphate buffer 50 mM at pH = 4 and pH = 7. The effect of methanol has also been studied when used as an organic modifier. Different voltages and electrokinetic conditions have been used to optimize the separation. Twenty Kv voltage and 10 Kv for 3 sec for injection have been found to be reliable conditions of separation at pH 4 without organic modifier. PMID- 12757122 TI - Monitoring the unfolding and refolding of creatine kinase by capillary zone electrophoresis. AB - Creatine kinase (ATP: creatine N-phosphotransferase; EC 2.7.3.2) plays a key role in the energy transport, muscle contraction, and reproduction of adenosine triphosphate (ATP). The activity of the enzyme is dependent on the correct folding of the peptide. We observed the unfolding and refolding processes of the creatine kinase of rabbit muscle, and concluded that traditional electrophoresis technology is unsuitable for the transient folding intermediates formed during protein unfolding and refolding. Capillary zone electrophoresis with diode array detection was used to monitor the unfolding and refolding of rabbit creatine kinase under different pretreatments and experimental conditions. This technique provides a simple, sensitive, and rapid approach to protein unfolding and refolding. PMID- 12757123 TI - Effect of nonaqueous buffer modifiers on the capillary electrophoresis-mass spectrometry analysis of peptides. AB - The effect of the addition of nonaqueous modifiers in capillary electrophoresis at varying concentrations was determined using synthetic peptides as model compounds. The electroosmotic flow is compared in each of the binary solvent systems and, in all cases, an increase in the concentration of organic solvent resulted in a decrease in the electroosmotic flow. The electrophoretic mobilities of each peptide were calculated and, in the case of acetonitrile, little or no change was observed in the peptide mobilities. In general, a decrease in the electrophoretic mobilities of the peptides was observed for increasing concentrations of methanol and N-methylformamide. The compatibility of each modifier at 25% (vol/vol) for use with mass spectrometric detection was evaluated. From our data, the addition of acetonitrile to the CE buffer appears to be the most compatible with mass spectrometry due to shorter analysis times and increased sensitivities in comparison to methanol and N-methylformamide. PMID- 12757124 TI - Large-volume sample stacking in nonaqueous capillary electrophoresis using alcoholic run buffers. AB - A straightforward method for stacking an extremely large volume of a sample solution containing weakly acidic organic compounds was developed. By using alcohols such as methanol, ethanol, and 1-propanol as run buffer solvents, the electroosmotic flow was suppressed, so that analyte anions could proceed to the outlet vial against the electroosmotic flow under a reverse voltage. This effect made large-volume sample stacking possible for large anions using a bare, fused silica capillary without intermediate polarity switching. The detection limits were in the low nanomolar range with conventional UV absorbance detection. The applicability of the technique to other organic solvents was also tested. PMID- 12757125 TI - Capillary electrophoretic assay of acyl-CoA hydrolase activity. AB - Acyl-CoA hydrolases catalyze the hydrolysis of fatty acyl CoA thioesters to free fatty acids and coenzyme A. These enzymes play an important role in the maintenance of cellular acyl-CoA and free CoASH pools and in the detoxification of nonphysiological metabolites. The assays most commonly used for acyl-CoA hydrolase quantitation are spectrophotometric and radioisotopic methods, both of which have limitations. In this study, capillary electrophoresis was used as an effective analytical technique to characterize rat liver peroxisomal acyl-CoA hydrolase reactivity using octanoyl-CoA as a substrate at different reaction conditions. The substrate and product of acyl-CoA hydrolase were identified by their migration times and quantitated using the peak areas. The enzyme activity exhibited a typical Michaelis-Menten pattern with increasing octanoyl-CoA concentration. The apparent Km and Vmax of octanoyl-CoA hydrolysis were determined using the enzyme activity at varying substrate concentrations. The rate of hydrolysis of octanoyl-CoA with increasing enzyme concentration appeared as a hyperbolic plot. The enzyme activity became elevated with increasing incubation time, showing the highest activity at 20 min, after which it started to decrease as the incubation time increased. Thus, capillary electrophoresis has been shown to be an effective, rapid, and reproducible method for the characterization of acyl-CoA hydrolase. PMID- 12757126 TI - Capillary electrophoresis separation of autocondensation glycation products of glucosamine. AB - Glucosamine nonenzymatically forms autocondensation glycation products under physiological conditions. Many studies have reported the effectiveness of oral doses of glucosamine alone or in combination with the galactosamine containing chondroitin in treating osteoarthritis. However, none of these studies has considered whether it is the glucosamine itself and/or one or more of its autocondensation products that exert this effect. A capillary electrophoresis method was developed to monitor the nonenzymatic formation of autocondensation glycation products of glucosamine, galactosamine, and mannosamine under physiological conditions. Major components were detected and separated by CE with a UV detector. The effects of concentration and incubation time on product species were determined. The method described is simple, rapid, and effective. PMID- 12757127 TI - Confidence and competence of recent veterinary graduates--is there a problem? PMID- 12757128 TI - An ethicist's commentary on veterinarians treating unowned animals and euthanizing unwanted animals. PMID- 12757129 TI - Employer and new graduate satisfaction with new graduate performance in the workplace within the first year following convocation from the Ontario Veterinary College. AB - Mailed questionnaires administered to employers of graduates and to graduates of the Ontario Veterinary College in 2000 and 2001, 7 to 10 months after convocation, surveyed new graduate performance in the workplace. Proficiency at 9 species-specific (in 4 practice contexts) and 7 nonspecies-specific clinical activities were rated as "high," "some," or "low." Fifteen nonvocation-specific attributes, reflecting interpersonal, communication, and business skills, and the new graduate's competence to do his/her job were rated as "very good," "good," or "poor." Ninety or more percent of employers reported "high" to "some" proficiency in 8/9, 5/9, 3/9, and 1/9 activities relative to small animal, food animal, equine, and exotic animal practice, respectively, and in 5/7 nonspecies-specific clinical activities. Ninety or more percent of employers assessed workplace proficiency as "very good" to "good" in 13/15 nonvocation-specific work skills and overall competence to do the job for which the new graduate had been hired. PMID- 12757130 TI - The effect of perineural anesthesia on infrared thermographic images of the forelimb digits of normal horses. AB - Infrared thermography is an imaging modality gaining popularity as a diagnostic aid in the evaluation of equine lameness. Anecdotal reports of skin hyperthermia induced by local anesthesia, detected by thermography, have been made; however, no controlled studies have been reported. The purpose of this study was to examine the effects of perineural anesthesia on infrared thermographic images of the forelimb digits in normal horses. After environmental acclimation, infrared thermographs were made at intervals of 0, 5, 10, 15, 30, and 45 min from administration of mepivacaine hydrochloride or phosphate buffered saline in 6 adult horses with no clinical evidence of abnormality of the forelimb digits. The mean limb surface temperatures were compared by 2-factor ANOVA. Results indicated no significant difference between treatments, time after injection, or an interaction of time and treatment. Infrared thermographic imaging apparently can be performed within 45 min of perineural mepivacaine hydrochloride anesthesia without risk of artifactual changes in limb surface temperature. PMID- 12757131 TI - Factors influencing the development of jugular thrombophlebitis in cattle and comparison of 2 types of catheter. AB - Two studies were conducted to evaluate the factors associated with the development of thrombophlebitis in cattle following intravenous catheterization of the jugular vein. In study 1, 20 healthy animals were catheterized with 2 different types of catheter (polytetrafluoroethylene (PTE) versus polyvinylchloride (PVC)) for a period of 120 hours. In study 2, 50 dairy cows referred for treatment to the Large Animal Veterinary Teaching Hospital at the University of Montreal were catheterized with a commercial PTE catheter, using a standardized technique, for a period varying from 3 to 6 days, and inherent risk factors were evaluated. A clinical and echographical evaluation of the 2 groups in study 1 demonstrated a higher frequency of thrombosis in the PTE group than in the PVC group. In study 2, the primary factors associated with the development of thrombophlebitis in sick cows were the experience of the manipulator and the severity of the disease. PMID- 12757133 TI - Evaluation of the California mastitis test to detect an intramammary infection with a major pathogen in early lactation dairy cows. AB - The purpose of this study was to evaluate the usefulness of the California mastitis test (CMT) to detect an intramammary infection caused by a major mastitis pathogen in early lactation cows. The gold standard used for comparison was bacteriological culture of single milk samples. The sensitivity (82.4%) and specificity (80.6%) of a positive CMT were highest on the 4th day of lactation. PMID- 12757132 TI - Effects of moderate to severe osteoarthritis on canine thyroid function. AB - Several nonthyroidal illnesses in euthyroid dogs can affect the results of thyroid function testing, making interpretation of the results more difficult with an increased risk of overdiagnosing hypothyroidism. The purpose of this study was to evaluate the effect of chronic, moderate to severe, osteoarthritis on canine thyroid function. Ninety-six, healthy, client-owned dogs, 65 of which were suffering from moderate to severe osteoarthritis and 31 euthyroid dogs without any physical evidence of osteoarthritis, were used in this study. Blood samples were collected to evaluate serum basal total thyroxine (TT4), free thyroxine (FT4), and thyrotropin (TSHc) concentrations. Basal serum TT4 concentration was not affected by osteoarthritis in dogs. Mild, but statistically significant, differences were noticed in FT4 and TSHc concentrations among the 2 groups. However, this had limited clinical relevance, since virtually all values were within their reference range, and no dogs would have been misdiagnosed as hypothyroid. Therefore, based on the results of our study, osteoarthritis does not need to be considered a factor influencing thyroid function evaluation in dogs. PMID- 12757134 TI - Long-term prevention of estrus in the bitch and queen using chlormadinone acetate. AB - Estrus was prevented with weekly oral administration of 2 mg chlormadinone acetate for 2.0 to 9.8 y in bitches and queens. Abnormalities, including mammary or uterine disorders, or both, were noted in 7 out of 14 bitches and 9 out of 24 queens during this long-term treatment. PMID- 12757136 TI - Gastric ulceration in an equine neonate. AB - A 24-hour-old colt presented with clinical signs consistent with gastric ulceration. Treatment was initiated with a histamine type-2 receptor antagonist and clinical signs resolved. Gastroscopy at 16 d confirmed the presence of a gastric ulcer. Although gastric ulceration is common in foals, it is rarely reported in foals this young. PMID- 12757135 TI - Juvenile cellulitis in a puppy. AB - An 8-week-old, male Labrador retriever presented for acute onset of left hind limb lameness. This rapidly progressed to juvenile cellulitis, characterized by dermatitis of the face, otitis externa, regional lymphadenopathy, lethargy, and depression. The puppy made a full recovery on glucocorticoid therapy. PMID- 12757137 TI - Diagnostic ophthalmology. Congenital lens luxation and secondary glaucoma. PMID- 12757138 TI - When change in the self is mistaken for change in the world. AB - The authors examined whether and when changes in the self lead to mistaken assessments that the world has changed. Survey data revealed that: personal changes in respondents (e.g., parenthood, financial change) were positively correlated with their assessments of various social changes (e.g., crime rates, freedom). Experimental data provided converging evidence. Experimentally induced change in knowledge influenced participants' perceptions of change in an author's writing style from one decade to the next (Study 3). Bringing self-change to participants' attention attenuated their judgments of change in the world when they had sufficient cognitive resources to consider how such self-changes might affect their perceptions (Studies 4-6). Discussion highlights how such misattributions of change contribute to the pervasive belief in societal decline. PMID- 12757139 TI - Malicious pleasure: schadenfreude at the suffering of another group. AB - Two studies examined intergroup schadenfreude--malicious pleasure at an out group's misfortune. Study 1 showed that schadenfreude regarding a German loss in soccer was increased by interest in soccer and threats of Dutch inferiority. The effect of inferiority threat was especially strong for participants less interested in soccer; the more interested showed relatively high schadenfreude. Study 2 replicated these effects by showing a similar pattern of schadenfreude regarding losses by Germany and Italy in another setting. However, schadenfreude toward legitimately superior Italy was lower when a norm of honest and direct expression was made salient to participants lower in soccer interest. These results establish schadenfreude as an emotion that is moderated by the salient dimensions of particular intergroup relations. PMID- 12757140 TI - The role of intent and harm in judgments of prejudice and discrimination. AB - Four experiments examined how an actor's intent and the harm experienced by a target influence judgments of prejudice and discrimination. The presence of intent increased the likelihood that participants judged an actor as prejudiced and the actor's behavior as discriminatory. When intent was uncertain, harm influenced judgments of the behavior, which in turn influenced judgments of the actor, and participants were more cautious in their judgments about an actor than an actor's behavior. Harm also played a stronger role in targets' than observers' judgments. Understanding the role of intent and harm on perceptions of prejudice can help explain variations in targets' versus observers', and possibly targets' versus actors', judgments of discrimination and prejudice. PMID- 12757141 TI - Exposure to violent media: the effects of songs with violent lyrics on aggressive thoughts and feelings. AB - Five experiments examined effects of songs with violent lyrics on aggressive thoughts and hostile feelings. Experiments 1, 3, 4 and 5 demonstrated that college students who heard a violent song felt more hostile than those who heard a similar but nonviolent song. Experiments 2-5 demonstrated a similar increase in aggressive thoughts. These effects replicated across songs and song types (e.g., rock, humorous, nonhumorous). Experiments 3-5 also demonstrated that trait hostility was positively related to state hostility but did not moderate the song lyric effects. Discussion centers on the potential role of lyric content on aggression in short-term settings, relation to catharsis and other media violence domains, development of aggressive personality, differences between long-term and short-term effects, and possible mitigating factors. PMID- 12757142 TI - Do not prime hawks with doves: the interplay of construct activation and consistency of social value orientation on cooperative behavior. AB - Low and high consistent pro-socials and pro-selfs were primed with neutral, morality, or might concepts in mixed-motive situations. The authors expected participants' social value orientation to influence cooperative behavior among (a) high consistent individuals in all prime conditions and (b) low consistent individuals in the neutral prime condition only. The authors also expected the primes to influence cooperative behavior more among low than high consistent individuals. Four experiments using supra-liminal (Experiments 1, 2, and 4) or subliminal (Experiment 3) priming and 2-person (Experiments 1-3) or N-person (Experiment 4) social dilemmas partially supported these initial predictions. One intriguing exception was that morality primes reduced cooperation among high consistent pro-selfs. Experiments 2-4 allowed testing for the potential role of expectations in shaping participants' cooperative behavior. PMID- 12757143 TI - Retrospection in social dilemmas: how thinking about the past affects future cooperation. AB - This article reports 2 studies investigating the effects of retrospective thought on future cooperation in social dilemmas. Some general theories of cooperation presume, but have not tested, whether retrospection has impact: People may think about the choices they could have made instead, realize that cooperation would have produced larger outcomes, and change their strategy as a result. Across both studies, the authors show that rate of future cooperation is directly related to the number of best-case scenarios and inversely related to the number of worst case scenarios generated. The 2nd study also shows that the number and type of retrospective thoughts generated can be predicted from the person's social value orientation. PMID- 12757144 TI - Male honor and female fidelity: implicit cultural scripts that perpetuate domestic violence. AB - Two studies explored how domestic violence may be implicitly or explicitly sanctioned and reinforced in cultures where honor is a salient organizing theme. Three general predictions were supported: (a) female infidelity damages a man's reputation, particularly in honor cultures; (b) this reputation can be partially restored through the use of violence; and (c) women in honor cultures are expected to remain loyal in the face of jealousy-related violence. Study 1 involved participants from Brazil (an honor culture) and the United States responding to written vignettes involving infidelity and violence in response to infidelity. Study 2 involved southern Anglo, Latino, and northern Anglo participants witnessing a "live" incident of aggression against a woman (actually a confederate) and subsequently interacting with her. PMID- 12757145 TI - When forgiving enhances psychological well-being: the role of interpersonal commitment. AB - The present research addresses the question of when and why forgiving might enhance psychological well-being. The authors predict that forgiving is associated with enhanced well-being but that this association should be more pronounced in relationships of strong rather than weak commitment. This hypothesis received good support in Studies 1-3. Studies 2 and 3 addressed the issue of why forgiving might be associated with psychological well-being, revealing that this association was reduced after controlling for psychological tension (i.e., a psychological state of discomfort due to conflicting cognitions and feelings). Study 4 revealed that in the context of marital relationships, tendencies toward forgiving one's spouse exhibited a more pronounced association with psychological well-being than did tendencies to forgive others in general. PMID- 12757146 TI - Narcissism, sexual refusal, and aggression: testing a narcissistic reactance model of sexual coercion. AB - Laboratory analog studies investigated the theory that narcissism and reactance contribute to causing rape. In Study 1, narcissism correlated positively with rape-supportive beliefs and negatively with empathy for rape victims. In Study 2, narcissists reported more enjoyment than other men of film depictions that presented consensual, affectionate activity followed by rape (but not in response to either affection or rape alone). In Study 3, narcissists were more punitive than other men toward a female confederate who refused to read a sexually arousing passage aloud to them. PMID- 12757147 TI - Development of personality in early and middle adulthood: set like plaster or persistent change? AB - Different theories make different predictions about how mean levels of personality traits change in adulthood. The biological view of the Five-factor theory proposes the plaster hypothesis: All personality traits stop changing by age 30. In contrast, contextualist perspectives propose that changes should be more varied and should persist throughout adulthood. This study compared these perspectives in a large (N = 132,515) sample of adults aged 21-60 who completed a Big Five personality measure on the Internet. Conscientiousness and Agreeableness increased throughout early and middle adulthood at varying rates; Neuroticism declined among women but did not change among men. The variety in patterns of change suggests that the Big Five traits are complex phenomena subject to a variety of developmental influences. PMID- 12757148 TI - Emotional convergence between people over time. AB - The authors propose that people in relationships become emotionally similar over time--as this similarity would help coordinate the thoughts and behaviors of the relationship partners, increase their mutual understanding, and foster their social cohesion. Using laboratory procedures to induce and assess emotional response, the authors found that dating partners (Study 1) and college roommates (Studies 2 and 3) became more similar in their emotional responses over the course of a year. Further, relationship partners with less power made more of the change necessary for convergence to occur. Consistent with the proposed benefits of emotional similarity, relationships whose partners were more emotionally similar were more cohesive and less likely to dissolve. Discussion focuses on implications of emotional convergence and on potential mechanisms. PMID- 12757149 TI - Relationship between coping styles and perceptual asymmetry. AB - This study examined the relationship between coping styles and hemispheric asymmetry, on the basis of prior evidence of reduced posterior right hemisphere (RH) activity in depression, and the relationship between ruminative coping and depression. Two samples of undergraduates (N = 170) completed chimeric faces tasks and 2 measures of coping styles, the self-report Responses Styles Questionnaire and a behavioral choice task. In women but not in men, self reported rumination was associated with a decreased RH bias on the emotion-based chimeric task. In both genders, choosing to engage in an emotional task was associated with increased RH involvement. Results indicate that although brooding and dwelling on the negative may be associated with decreased RH involvement, openness to emotion may be associated with increased RH involvement. PMID- 12757150 TI - The role of racial identity in perceived racial discrimination. AB - This study examined the role that dimensions of racial identity play regarding the antecedents and consequences of perceived racial discrimination among African Americans. A total of 267 African American college students completed measures of racial identity, perceived racial discrimination, and psychological distress at 2 time points. After controlling for previous perceptions of discrimination, racial centrality was positively associated with subsequent perceived racial discrimination. Additionally, perceived discrimination was positively associated with subsequent event-specific and global psychological distress after accounting for previous perceptions of discrimination and distress. Finally, racial ideology and public regard beliefs moderated the positive relationship between perceived discrimination and subsequent distress. The results illustrate the complex role racial identity plays in the lives of African Americans. PMID- 12757151 TI - It takes two to mimic: behavioral consequences of self-construals. AB - The present studies demonstrated the moderation of self-construal orientation on mimicry. Recent research has indicated that an interdependent self-construal is associated with assimilation of the other to the self whereas an independent self construal is associated with minimizing the influence of others on the self (H. R. Markus & S. Kitayama, 1991; D. Stapel & W. Koomen, 2001). Therefore, the authors hypothesized that an interdependent self-construal would be associated with more mimicry than an independent self-construal. When self-construal orientations were experimentally primed, as in Studies 1 and 2, independent self construals produced less nonconscious mimicry than interdependent self construals. When self-construals were examined as cultural differences with either a chronically dominant independent (Americans) or interdependent (Japanese) construal of the self, these results were replicated. PMID- 12757154 TI - Morphological properties of zero-stress state in rat large intestine during systemic EGF treatment. AB - Systemic treatment with epidermal growth factor (EGF) induce growth of the large intestine. The aim of this study was to investigate the morphological properties early in the course of EGF-induced large intestinal growth. The effects of systemic EGF treatment on the morphological properties at the zero-stress state along the large intestine were investigated. EGF-treated rats and control rats were allocated into group with EGF treatment for 2, 4, 7, and 14 days (N = 6 for each EGF treatment group except N = 4 for the 14-day group). The controls had saline injected (N = 3 for each group). The excised large intestine was subdivided into four segments: the ascending colon, transverse colon, descending colon, and rectum. The length and weight of each segment was measured. The zero stress state was obtained by cutting rings of the large intestine radially, and the opening angle was measued on video images. The thickness and cross-sectional area of the mucosa and muscle layers, and the inner and outer circumference were measured. The total colon length did not increase, whereas the weight of the large intestine, muscosal thickness and mass, and inner and outer circumference increased significantly (P < 0.05). The increase was most prominent in the proximal colon after 7 days of EGF treatment. Later no further morphological changes were observed, except for a decrease in mucosal thickness in most segments and in mucosal cross-sectional area in the descending colon. In the controls and during the first week of EGF treatment, the opening angle was approximately 100 degrees. After 14 days the opening angle increased significantly in the ascending and transverse colon to approximately 172 and 135 degrees (P < 0.05). In conclusion, this study demonstrates that systemic EGF treatment caused remodeling of the morphology of the zero-stress state in the large intestine in a time-dependent manner. The growth was most pronounced in the ascending and transverse colon and involved mainly the mucosal layer. PMID- 12757153 TI - Spasmolytic polypeptide-expressing metaplasia (SPEM) associated with gastric cancer in Iceland. AB - Recent studies have described a spasmolytic polypeptide-expressing metaplastic cell lineage (SPEM) in the gastric fundic mucosa associated with both chronic H. pylori infection and gastric adenocarcinoma. We investigated the association of SPEM both with early gastric adenocarcinoma and in biopsies taken from patients prior to diagnosis of cancer. Two cohorts were examined. First, gastric resections from 29 patients with early gastric cancer were examined. Second, biopsies taken from 18 patients prior to the diagnosis of gastric cancer were compared with their respective resection specimens as well as with control biopsies from a cohort of 19 patients diagnosed with gastritis without subsequent development of cancer. The presence of SPEM and intestinal metaplasia (IM) adjacent to and distant from the cancer was compared and spasmolytic polypeptide (SP) immunostaining within dysplastic/cancerous cells was identified. SPEM was present adjacent to cancer in all early cancer cases where the tumor was located in the body or at the body/antrum junction, and was present in the body mucosa distant from the cancer in 76% of cases. Intestinal metaplasia was found adjacent to the tumor in 76% of cases and in body sections in 52% of resections. SP immunostaining was noted within cancer cells in 62% of tumors, and within dysplastic cells in 76% of resections where dysplasia was present. SPEM was present in 82% of the biopsies obtained prior to the diagnosis of cancer, compared with only 37% in the gastritis cohort. IM was present in only 57% of biopsies. In conclusion, SPEM is strongly associated with early gastric cancers and is observed in gastric biopsies prior to the development of cancer. In addition, early gastric cancers demonstrated a high incidence of SP expression. These results suggest that SPEM merits consideration as an important pre neoplastic gastric lesion. PMID- 12757152 TI - Overlap syndromes of autoimmune hepatitis: what is known so far. AB - The pattern of some autoimmune hepatitis can be difficult to classify, sometimes due to the overlap of these with primary biliary cirrhosis, primary sclerosing cholangitis and chronic viral hepatitis. The etiology of these variant forms remains unclear. The distinction among the overlap syndromes poses different problems both of prognosis and therapeutic approach. Presently, the utility of the scoring system devised and revised by the International Autoimmune Hepatitis Group regarding these cases is under discussion. Histological examination seems to be an important tool, but often the result does not help in defining a correct diagnosis. To date, the overlap syndromes can be classified at an intermediate level between cholestatic forms of autoimmune hepatitis or hepatic forms of cholestatic syndromes, but it cannot be excluded that the syndromes represent independent disorders. PMID- 12757155 TI - Vascular involvement in pancreatic neoplasm: a comparison between spiral CT and DSA. AB - The principal criterion for resectability of pancreatic carcinoma is the assessment of vascular involvement. In a prospective evaluation the ability of Spiral CT Angiography (CTA) to detect vascular involvement in 50 patients with pancreatic carcinoma, was proved; DSA was performed later in all patients. In 20 patients, without vascular involvement, a complete concordance was obtained. Of 30 patients with vascular involvement, there was complete concordance between CTA and angiography in 22 patients and discordance in 8 patients. CTA was superior in 2 cases with periadventitial infiltration and in 5 patients with splenoportal confluence thrombosis. DSA was superior in 1 case with infiltration of the superior mesenteric vein. After surgical evaluation, sensitivity of CTA and DSA was 97% and 77%, respectively, and the negative predictive values were 95% and 74%. As compared to DSA, CTA is more rapid and less invasive and can be considered the modality of choice for preoperative work-up of pancreatic neoplasm. PMID- 12757156 TI - Overexpression of pancreatitis-associated protein (PAP) in human pancreatic ductal adenocarcinoma. AB - Pancreatitis-associated protein (PAP) is almost absent in normal pancreas, but is strongly induced in acute pancreatitis. PAP mRNA is also expressed in cancer cells, including pancreatic ductal adenocarcinoma. However, the clinicopathological significance of PAP in human pancreatic cancer is not clear. We examined PAP expression in pancreatic tissues from individuals with pancreatic ductal adenocarcinoma using immunohistochemistry. PAP was overexpressed in 79% (30 of 38) of pancreatic ductal adenocarcinoma, 19% (7 of 36) of chronic pancreatitis, and 29% (2 of 7) of mucinous cystadenoma. PAP was found in malignant ductular structures in pancreatic carcinomas as well as in benign proliferating ductules and acinar cells in chronic pancreatitis. It was not expressed in normal pancreas. The incidence of PAP overexpression was significantly higher in pancreatic cancer than in the other pancreatic diseases (P < 0.01). PAP overexpression was significantly correlated with nodal involvement, distant metastasis (P < 0.05), and short survival (P < 0.01) in pancreatic cancer. These results suggest that overexpression of PAP in human pancreatic ductal adenocarcinoma indicates tumor aggressiveness. PMID- 12757157 TI - Verrucous carcinoma of the esophagus: clinicopathophysiologic features and treatment of a rare entity. PMID- 12757159 TI - Helicobacter pylori cytotoxins and tyrosine phosphatase functions. PMID- 12757158 TI - Helicobacter pylori infection and high dietary salt independently induce atrophic gastritis and intestinal metaplasia in commercially available outbred Mongolian gerbils. AB - Risk factors for development of gastric adenocarcinoma include high dietary salt and Helicobacter pylori infection. Few animal models exist for the laboratory investigation of these factors. We examined gastric pathology resulting from H. pylori infection and high dietary salt as independent variables in commercially available, outbred Mongolian gerbils. Gastric adenocarcinoma and its precursor lesion, intestinal metaplasia, have been previously reported in inbred Mongolian gerbils (MGS/Sea) infected either with clinical isolates of H. pylori or with the strain ATCC 43504. In contrast, we utilized outbred gerbils [Crl:(MON)] infected with the Sydney strain of H. pylori. After 37 weeks, five of five infected animals had atrophic gastritis and intestinal metaplasia. These lesions were similar in description and time of appearance to the lesions reported in inbred gerbils. Atrophic gastritis and intestinal metaplasia also developed in six of six uninfected, outbred gerbils fed a 2.5% salt diet for 56 weeks. In contrast to the H. pylori-infected animals, these lesions were present without concurrent gastric inflammation. The outbred Mongolian gerbil therefore provides an excellent animal model for the study of several gastric cancer risk factors. PMID- 12757160 TI - Effect of altering gastric emptying on postprandial plasma glucose concentrations following a physiologic meal in type-II diabetic patients. AB - The purpose of this study was to determine the effects of altering gastric emptying on postprandial plasma glucose concentration after a physiologic meal in patients with type II diabetes mellitus (T II DM). Nine T II DM patients underwent a double-blind, randomized, three-way crossover study, receiving erythromycin 200 mg, morphine 8 mg, or normal saline (placebo) intravenously prior to ingestion of a radiolabeled, dual-isotope, solid-liquid meal. Gastric emptying of solids and liquids and serial plasma glucose, glucagon, and serum insulin concentrations were measured at baseline and for 5 hr after meal ingestion. Erythromycin accelerated and morphine delayed solid- and liquid-phase gastric emptying compared to placebo (P < 0.05). During the first hour, the postprandial plasma glucose concentrations were higher after erythromycin (P < 0.05) and lower after morphine (P < 0.05) compared to placebo. The peak postprandial plasma glucose concentration was higher after erythromycin (P = 0.05) and lower after morphine (P < 0.05) compared to placebo. In conclusion, pharmacologic acceleration of gastric emptying resulted in higher postprandial glucose concentrations, while delaying gastric emptying resulted in lower postprandial glucose concentrations after a physiologic meal in T II DM. These results suggest that administration of opiate analgesics or prokinetic agents to diabetic patients may alter glucose control. Modifying gastric emptying may be helpful in achieving glucose control in T II DM. PMID- 12757161 TI - Vector analysis of electrogastrography during motion sickness. AB - Although the dominant frequency at 3 cpm of electrogastrography (EGG) is reported to shift to a higher frequency during motion sickness (MS), it is unclear whether the normal slow wave (NSW) disappears or not. The authors investigated changes in NSW using vector analysis of EGG. Fourteen subjects were exposed to a Coriolis stimulation to evoke MS. EGG was recorded from two sets of bipolar leads, placed perpendicular to each other representing x and y axes. Trajectories for each frequency were drawn on the x-y plane. The amplitude and phase difference at NSW were compared before, during, and after the stimulus for each subject. In those with a change in phase difference, changes in NSW and tachygastria were negatively correlated (P = -0.048), whereas in those without a change, they were not correlated. This indicated two different kinds of tachygastria due to MS: tachygastria with and without a change in NSW. PMID- 12757162 TI - Origins and patterns of spontaneous and drug-induced canine gastric myoelectrical dysrhythmia. AB - The aim of this study was to investigate the characteristics and orientation of gastric dysrhythmia using multichannel serosal recordings in dogs. Ten dogs chronically implanted with four to eight pairs of electrodes were studied. Gastric slow waves were recorded in four sessions: postsurgical and after atropine, vasopressin, and glucagon. A total of 554.7 min of bradygastria, 201 min of tachygastria and 22.3 min of arrhythmia were observed in the recordings. The majority of bradygastria (80.5 +/- 9.4%) originated in the proximal stomach (P < 0.04, vs other locations) and propagated all the way to the distal antrum. In contrast, tachygastria mainly originated in the distal antrum (80.6 +/- 8.8%) (P < 0.04, vs other locations) and propagated partially or all the way to the proximal stomach. Dysrhythmia appeared intermittently with normal gastric slow waves. In all recordings, normal slow waves were present 38.0 +/- 5.3% of the time, while bradygastria, tachygastria, and arrhythmia were present 35.9 +/- 5.3%, 23.0 +/- 1.6%, and 2.4 +/- 0.5% of the time, respectively. The prevalence of dysrhythmia was highest in the distal antrum (80.4%) (P < 0.01) and lowest in the proximal part of the stomach. In conclusion, tachygastria originates from an ectopic pacemaker in the distal antrum. It may completely or partially override the normal slow waves. Bradygastria is attributed to a decrease in the frequency of the normal pacemaker in the corpus. The prevalence of gastric dysrhythmia is different in different locations of the stomach and is highest in the distal antrum. PMID- 12757163 TI - Influence of gender, parity, and caloric load on gastrorectal response in healthy subjects: a barostat study. AB - The gastrocolonic response consists of a prompt increase in colonic tone after a meal. With a barostat and a high compliant air-filled bag, it is possible to measure rectal tone by recording changes in volume at a constant intrabag pressure. The aim of this study was to evaluate the gastrorectal response in males and females as well as the effect of different caloric loads on the gastrorectal response. In 33 volunteers a barostat procedure during basal conditions and after a 600-kcal meal was performed. In 26 volunteers the procedure was repeated with a 1000-kcal meal. A meal response was defined as a decrease in volume of more than 10%. Phasic volume events (PVE) were defined as a 10% decrease in volume of 15-60 sec duration. After a 600-kcal meal, the decrease in volume after 1 hr was 28 +/- 7% (mean +/- SEM, P < 0.001). A meal response was found in 64% of the subjects. Parous females had a diminished meal response compared with nulliparous females (2 +/- 5% and 48 +/- 11%, P < 0.001). After the 600-kcal meal, PVEs increased from 3 to 10/hr (P = 0.001). In the 26 subjects, volume decrease was 40 +/- 9% after the 1000-kcal meal and 20 +/- 7% after the 600-kcal meal (P = 0.28). In the high-calorie meal, 18 subjects (69%) had a response versus 14 (54%) in the low-calorie meal (NS). Enhancing the caloric load of the meal did not increase the amounts of PVEs. In conclusion, a gastrorectal response occurs in 64% of the healthy subjects after a 600-kcal meal. The gastrorectal response can be measured to a similar extent in men and nulliparous women; however, the response is significantly impaired in parous women. This is possibly due to neurogenic damage during childbirth. Increasing the caloric load did not increase the gastrorectal response. Therefore, to study gastrorectal meal response with the barostat, a meal of 600-kcal is sufficient and a correction for parity should be made when results are compared. PMID- 12757164 TI - EGG data. PMID- 12757166 TI - Vitamin D3 in patients with various grades of chronic pancreatitis, according to morphological and functional criteria of the pancreas. AB - There are still too few conclusive reports about conspicuous vitamin D deficiency in patients with chronic pancreatitis, or any connection of the deficiency to the severity of the disease. Between October 1999 and September 2000, we investigated 42 patients at an average age of 53 years, suffering from chronic pancreatits, as well as 20 healthy male controls at an average age of 49 years. Serum levels of D3 vitamins, 1,25-(OH)2-vitamin D3 and 25-(OH)-vitamin D3, as well as the concentration of fecal elastase 1 were determined in patients and controls. Furthermore, the severity of chronic pancreatitis in patients was determined via endoscopic retrograde cholangiopancreatography (ERCP) into 3 grades, based on the Cambridge classification. Elastase 1 in feces revealed sensitivities of 14%, 87%, and 95% for Cambridge-grades I, II, and III, respectively, and correlated significantly with this classification of severity of chronic pancreatitis (P < 0.01). In patients with Cambridge-grade II and III 1,25-(OH)2-D3 was markedly decreased (26.7 +/- 7.7 pg/ml and 27.6 +/- 9.0 pg/ml) compared to those with Cambridge-grade I (38.0 +/- 10.5 pg/ml; between I and II P = 0.027, between I and III P = 0.033). 25-(OH)-D-3 did not differ significantly within the various Cambridge-grade groups (P = 0.07). Nevertheless, vitamin D3 and fecal elastase 1 in patients correlated significantly (P < 0.01) and, compared to controls, both were extremely low (means in patients: fecal elastase 1 140.7 +/- 75.7 microg/g, 1,25-(OH)2-D3 29.9 +/- 9.5 pg/ml, 25-(OH)-D3 26.7 +/- 9.7 nmol/liter; controls: fecal elastase 1 694.9 +/- 138.6 microg/g, 1,25-(OH)2-D3 67.5 +/- 4.3 pg/ml, 25 (OH)-D3 69.5 +/- 13.5 nmol/liter). The amounts of both D3 vitamins in patients were significantly lower when the content of fecal elastase 1 was under 200 microg/g compared to the others [for 1,25-(OH)2-D3 P < 0.01, for 25-(OH)-D3 P < 0.05]. Therefore, ERCP and fecal elastase 1 verify the severity grade of a chronic pancreatitis, and thus show a vitamin D3 deficiency, depending on the progress of the disease. There seems to be a connection between inflammatory pancreas destruction (Cambridge classification), exocrine insufficiency (fecal elastase 1), and perhaps even the characteristics of sterol-binding of pancreatic elastase 1, which seems to be relevant for vitamin D supply. PMID- 12757167 TI - Cholestatic hepatitis induced by Epstein-Barr virus infection in an adult. AB - Liver involvement is nearly universal in healthy persons with Epstein-Barr Virus (EBV) infection-induced infectious mononucleosis. It is usually mild, undetected clinically and resolves spontaneously. Jaundice is distinctly uncommon and may reflect either more severe hepatitis or an associated hemolytic anemia. Cholestatic hepatitis due to EBV infection is infrequently reported and may pose a diagnostic quandary. We describe a patient who presented with jaundice and a markedly elevated serum alkaline phosphatase level due to serologically confirmed acute infection with EBV. Imaging studies excluded biliary obstruction. Symptoms and laboratory abnormalities resolved spontaneously. EBV infection should be included in the differential diagnosis of cholestatic hepatitis in adults. PMID- 12757165 TI - Acid-induced mesenteric hyperemia in rats: role of CGRP, substance P, prostaglandin, adenosine, and histamine. AB - Intraduodenal acidification produces a mesenteric hyperemia that is mediated in part by mucosal capsaicin-sensitive afferent nerves and the bradykinin B2 receptor in anesthetized rats. We hypothesized that novel mechanisms mediated by substance P, adenosine, and histamine1 receptors are involved. Confirmation of a role for calcitonin gene-related peptide (CGRP) but not endogenous prostaglandin was also sought. In study 1, vehicle or antagonists (CGRP(8-37), CP 96345) was administered intravenously. Capsaicin or acid was administered intraduodenally, followed by intravenous CGRP or substance P. In study 2, pretreatments included indomethacin, 8-phenyltheophylline, pyrilamine, or the respective vehicles. Acid was then administered intraduodenally. In both studies, superior mesenteric artery blood flow was monitored. In study 1, the antagonists significantly attenuated capsaicin- and acid-induced mesenteric hyperemia. In study 2, the pretreatments did not alter acid-induced hyperemia. The data confirmed the role of CGRP and indicated for the first time an involvement for substance P in acid induced mesenteric hyperemia. PMID- 12757168 TI - TIPS for management of refractory ascites: response and survival are both unpredictable. AB - Refractory ascites is a serious complication of advanced cirrhosis with a 1-year transplant-free survival of 20-50%. The aim of our study was to investigate the short- and long-term effects of transjugular intrahepatic portosystemic shunt (TIPS) in the management of refractory ascites. In all 65 patients (39 M, 26 F; Child B 55%, Child C 45%, mean MELD score 14.8 +/- 6.6) with liver disease (alcoholic 40%, cryptogenic 20%, HCV 14%, others 26%) and refractory ascites were included in this study. Forty-eight (74%) patients had no signs of hepatic encephalopathy (HE), 16 (24%) had mild and 1 (2%) had moderate HE before TIPS; 28 (43%) had mild (> 1.2 and < 2.4 mg/dl) and 6 patients (9%) had moderate (> 2.4 mg/dl) renal dysfunction. Mean follow-up was 55.5 +/- 70.2 weeks. Treatment success, defined as complete response, partial response, and no response, and survival was determined at 3 weeks, and 3, 6, 12, 24, and 36 months after TIPS. TIPS was successful in all patients. Mean portal venous pressure gradient improved significantly after TIPS (24 +/- 8 to 10 +/- 4). During follow-up, 40 (58%) patients died and 17 (27%) patients had liver transplantation (OLT); 20 (31%) patients had 38 shunt revisions due to lack of initial response or recurrence of ascites. The response was assessed in patients who were alive, without OLT, at each time point. Complete response was seen in 10%, 23%, 17%, 11%, 22% and 33%; partial response was seen in 46%, 46%, 40%, 44%, 28%, and 8%; and no response was seen in 44%, 31%, 43%, 41 %, 39%, and 50% at 3 weeks, and 3, 6, 12, 24, and 36 months respectively. There were no pre-TIPS variables that could predict the response at 3 weeks, 3 months, or 6 months. Mild HE was seen in 8 (12%) patients and severe HE was seen in 16 (25%) immediately after TIPS. The mortality at 3 weeks, and 3, 6, 12, 24, and 36 months was 26%, 38%, 46%, 51%, 57%, and 58%, respectively. Three-week (P = 0.01) and 3-month (P = 0.04) mortality was higher in Child C patients compared to Child B. However, there were no independent predictors of survival on multivariate analysis at 3 or 6 months. Child-Pugh score 3 weeks after TIPS was a strong predictor of mortality. In conclusion, in patients with refractory ascites, TIPS was associated with a high mortality and morbidity. The response and the mortality were both unpredictable on the basis of pretransplant variables. PMID- 12757169 TI - Spontaneous closure of intrahepatic portovenous shunt in a noncirrhotic patient with recurrent encephalopathy. AB - A 70-year-old man was admitted to our hospital with recurrent encephalopathy. Liver function tests, abdominal computerized tomography, ultrasonography, angiography, and laparoscopy revealed an intrahepatic portovenous shunt in a noncirrhotic liver. During follow-up, the intrahepatic portovenous shunt closed spontaneously. Subsequent liver function tests were markedly improved, with resolution of the patient's previously disturbed consciousness. In the elderly, intrahepatic portovenous shunt that can be managed with conservative therapy may spontaneously close, suggesting that management options should include watchful waiting. PMID- 12757170 TI - Hepatopulmonary syndrome in noncirrhotic portal hypertensive patients. AB - Hepatopulmonary syndrome has yet not been sufficiently assessed in noncirrhotic portal hypertension. The prevalence of hepatopulmonary syndrome was determined in 31 consecutive patients with noncirrhotic portal hypertension (19 idiopathic portal hypertension, 7 portal vein thrombosis, 5 congenital hepatic fibrosis) and 46 patients with liver cirrhosis. Contrast echocardiography was carried out in all patients. Macroaggregated albumin lung perfusion scans were performed in patients with positive contrast echocardiogram. Hepatopulmonary syndrome was detected in 5 (10.8%) cirrhotic and 3 (9.7%) noncirrhotic portal hypertensive patients (2 idiopathic portal hypertension, 1 portal vein thrombosis). All patients with hepatopulmonary syndrome had an increased shunt fraction (13-62%) and a decreased diffusion capacity of carbon monoxide (40-79%), and 7 of them were hypoxemic (PaO2, 31.6-69.8 mm Hg). These findings show that hepatopulmonary syndrome may occur in both liver cirrhosis and noncirrhotic portal hypertension and that portal hypertension is the predominant etiopathogenic factor related to hepatopulmonary syndrome. PMID- 12757171 TI - Identification of beta-subunit of bacterial RNA-polymerase--a non-species specific bacterial protein--as target of antibodies in primary biliary cirrhosis. AB - Several observations suggest that bacteria induce autoimmunity in primary biliary cirrhosis (PBC). Since no PBC-specific bacterial species could be identified, it can be speculated that the triggers are non-species-specific bacterial proteins. This hypothesis would imply that several or even all bacterial species can trigger PBC. Therefore, we investigated whether PBC exhibits immune reactions to non-species-specific bacterial antigens. Yersinia enterocolitica O3 was screened for the presence of proteins that were labeled by immunoblotting using PBC sera. We focused our investigations on a 160-kDa protein, which was further enriched and characterized by partial N-terminal amino acid sequencing. The prevalence of antibodies to this protein was determined by immunoblotting in a variety of diseases. The 160-kDa protein was identified as the beta-subunit of bacterial RNA polymerase, a highly conserved bacterial protein with a very high degree of sequence identity among all bacterial species. Antibodies to the beta-subunit of bacterial RNA polymerase were specific for this protein. Until now no mammalian protein could be found that cross-reacts with these antibodies. The prevalence of antibodies to the beta-subunit of bacterial RNA polymerase (ARPA) using the protein from Yersinia enterocolitica O3 (serum dilution 1:1000) was: healthy controls (HC, N = 101) 7.9%, primary biliary cirrhosis (PBC, N = 61) 32.8%, autoimmune hepatitis type 1 (AIH, N = 46) 26.1%, alcoholic liver cirrhosis (ALC, N = 44) 9.1%, Crohn's disease (CD, N = 38) 7.9%, ulcerative colitis (UC, N = 24) 8.3%, primary sclerosing cholangitis + UC (PSC/UC, N = 11) 0%, acute yersiniosis (Yers, N = 36) 19.4%, acute infection with Campylobacter jejuni (Camp, N = 10) 0%, acute Q-fever (QF, N = 16) 6.25%, chronic hepatitis C (HCV, N = 39) 7.7%, c ANCA-positive vasculitis (Vasc, N = 40) 15%, systemic lupus erythematosus (SLE, N = 28) 10.7%, and malaria tropica (MT, N = 24) 16.7%. There was no significant difference between PBC and AIH. The group of autoimmune liver diseases (PBC + AIH, N = 107, 29.9%) differed highly significantly from HC, chronic inflammatory bowel diseases (CD + UC + PSC/UC, N = 73, 6.8%), ALC, and HCV and also differed significantly (P = 0.01) from the group with bacterial and parasitic diseases (Yers + Camp + QF + MT, N = 86,13.95%) and from the group with Vasc + SLE (N = 68,13.2%). Testing of ARPA using the protein from E. coli yielded nearly identical results. In conclusion, an increased prevalence of antibodies to the beta-subunit of bacterial RNA polymerase, a highly conserved non-species-specific bacterial protein, can be found in primary biliary cirrhosis, but also in autoimmune hepatitis type I. These findings do not add an argument for a bacterial trigger of PBC. Rather, they suggest that ARPA belong to the pool of natural antibodies that are up-regulated in autoimmune liver diseases. PMID- 12757173 TI - Growth rate of primary single hepatocellular carcinoma: determining optimal screening interval with contrast enhanced computed tomography. AB - To determine the optimal screening interval for detecting small (< 20 mm) hepatocellular carcinoma (HCC) in a high-risk group using multiphase contrast enhanced computed tomography (CECT), we evaluated the growth rate of primary single HCC. Forty-nine primary single HCC cases were reviewed. CECT screening was performed more than two times preceding to the diagnosis in 29 cases, and HCC nodule was identified at least two times in 22 cases. The initial nodule sizes ranged between 3 and 30 mm. Doubling time of tumor volume ranged from 34.8 to 496.4 days, with a geometric mean of 93.5 days, and a 95% lower threshold value of 27.1 days. It means that HCC will not double in diameter within 3 months. Therefore CECT screening at intervals of 3 months will detect new nodules at 10 20 mm in size and CECT screening at intervals of longer than 3 months will detect new nodules but they might be larger than 20 mm in size. PMID- 12757172 TI - Idoxifene and estradiol enhance antiapoptotic activity through estrogen receptor beta in cultured rat hepatocytes. AB - Oxidative stress plays a causative role in the development of hepatic fibrosis and apoptosis. Estradiol (E2) is an antioxidant, and idoxifene is a tissue specific selective estrogen-receptor modulator. We have previously demonstrated that E2 inhibits hepatic fibrosis in rat models of hepatic fibrosis and that the actions of E2 are mediated through estrogen receptors (ERs). This study reports on the antiapoptotic role of idoxifene and E2, and the functions of ER subtypes ER-alpha and ER-beta in hepatocytes undergoing oxidative stress. Lipid peroxidation was induced in cultured rat hepatocytes with ferric nitrilotriacetate solution with idoxifene or E2. Oxidative stress-induced early apoptosis was linked to its ability to inhibit not only the expression of Bcl-2 and Bcl-XL but the production of antioxidant enzymes as well and to stimulate Bad expression. Hepatocytes possessed functional ER-beta, but not ER-alpha, to respond directly to idoxifene and E2. Idoxifene and E2 suppressed oxidative stress-induced reactive oxygen species generation and lipid peroxidation, and their antiapoptotic effects on the activation of activator protein-1 and nuclear factor-kappaB, the loss of antioxidant enzyme activity, and Bcl-2 family protein expression in early apoptotic hepatocytes were blocked by the pure ER antagonist ICI 182,780. Our results indicate that idoxifene and E2 could enhance antiapoptotic activity through ER-beta during oxidative damage in hepatocytes. PMID- 12757174 TI - Computed tomography-guided transarterial chemoembolization as the initial therapy for hepatocellular carcinoma: experience of 75 cases in a single institute. AB - We present the survival rates of 75 nonruptured hepatocellular carcinoma cases initially treated with computed tomography-guided transarterial chemoembolization in a single institute. The 1-, 3-, and 5-year survival rates were 93.9%, 74.7%, and 47.4% in 50 Child's A cases; 75.0%, 43.6%, and 6.8% in 20 Child's B cases; and 60.0%, 40.0%, 0.0% in 5 Child's C cases, respectively. The 1-, 3-, and 5-year survival rates of the 38 estimated resectable hepatocellular carcinoma cases (Child's A, tumors limited in a single lobe) were 94.7%, 82.0%, and 44.6%, respectively. The 1-, 3-, and 5-year survival rates of the 41 cases with estimated indication for percutaneous ethanol injection therapy (tumors less than 3 cm in diameter and three or fewer in number) were 96.8%, 84.6%, and 55.5% in 31 Child's A cases; and 90.0%, 46.7%, and 0% in 10 Child's B cases, respectively. In conclusion, computed tomography-guided transarterial chemoembolization is an excellent primary therapy for hepatocellular carcinoma. PMID- 12757176 TI - Oxidant-stimulated chloride secretion in rat jejunum in vitro is mediated by eicosanoids. AB - Specific effects of the cytotoxic secondary lipid oxidation product, 4 hydroxynonenal (10(-8)-10(-4) M), on intact sheets of rat jejunum were measured as changes in short circuit current (delta(I)sc) following cumulative addition to either the mucosal or serosal side, using the analogous aldehyde, nonenal, as reference. 4-Hydroxynonenal stimulated I(sc) from the serosal side (maximal delta(I)sc = 27.2 +/- 3.5 microA/cm2, mean +/- SEM, N = 32) while nonenal stimulated I(sc) primarily from the mucosal side (maximal delta(I)sc = 16.2 +/- 3.4 microA/cm2, N = 20). Inhibition by 100 microM bumetanide (4-hydroxynonenal: 88.9 +/- 3.0%, N = 6, p < 0.05, nonenal: 69.3 +/- 2.9%, N = 6, P < 0.05) indicated chloride secretion. Nonenal-induced delta(I)sc was inhibited (72.5 +/- 1.2%, N = 8, P < 0.05) by a combination of nordihydroguaiaretic acid (100 microM) and piroxicam (10 microM), while 4-hydroxynonenal-induced delta(I)sc, was abolished by piroxicam (N = 8, P < 0.001) and inhibited by 1 microM tetrodotoxin (69.8 +/- 9.7%, N = 6, P < 0.001). These data indicate that 4-hydroxynonenal stimulates chloride secretion mediated by prostaglandins and the enteric nervous system. The site of action (serosal) being opposite to the reference aldehyde. PMID- 12757175 TI - Combination therapy with interferon-alpha(2b), ribavirin, and amantadine in chronic hepatitis C nonresponders to interferon and ribavirin. AB - Standard therapies for the treatment of hepatitis C are ineffective in almost 50% of patients. Amantadine is an antiviral agent that may have activity against hepatitis C virus. In this pilot study, we evaluated the efficacy of a combination of interferon, ribavirin, and amantadine in patients with chronic hepatitis C who had previously failed 6-12 months of treatment with interferon and ribavirin. In this prospective open-label study, 23 patients were treated with a combination of interferon-alpha(2b) 3 million units subcutaneously three times per week, ribavirin 1000-1200 mg daily, and amantadine 100 mg twice daily for 6-12 months. Treatment was discontinued at 6 months if the patients had detectable HCV RNA by PCR. All patients were followed for 6 months after the completion of treatment. At the end of treatment, the biochemical response was 47% and the virological response was 30%. However, the rate of sustained virological response was only 13% (3/23). There were no unexpected side effects with triple therapy. In conclusion, triple therapy with interferon, ribavirin and amantadine resulted in a low sustained viral clearance in chronic hepatitis C patients who had previously failed interferon and ribavirin combination therapy. PMID- 12757177 TI - Diagnostic yield of duodenal aspirate for G. lamblia and comparison to duodenal mucosal biopsies. AB - Our objective was to examine the diagnostic yield of duodenal aspirates for Giardia in children and compare it to results of duodenal mucosal biopsies. The results of all duodenal aspirates submitted for direct parasite examinations over a 31-month period were reviewed, as were the histological results of duodenal mucosal biopsies from these patients. In all, 161 children (89 boys; age range 0.33-18 years) were included in the study. Giardia was identified in the duodenal aspirate of 5.6% (9/161) patients and on duodenal mucosal biopsies from all nine patients. In conclusion, the 5.6% diagnostic yield of duodenal aspirates for Giardia is higher than reported in a previous study of adult patients from a similar geographical region (0.7%). The detection of Giardia on duodenal mucosal biopsies from all patients with positive duodenal aspirates brings into question the utility and cost of the latter test. Duodenal aspirates for Giardia may be unnecessary if duodenal mucosal biopsies are obtained for histological examination. PMID- 12757178 TI - High antigliadin IgG titers in laboratory rabbits fed a wheat-containing diet: a model for celiac disease? PMID- 12757179 TI - Distribution of HLA class I alleles differs in celiac disease patients according to age of onset. AB - Celiac disease (CD) or gluten-sensitive enteropathy is strongly associated with HLA-DQ alleles; more than 95% of patients are DQB1*02. However, the uniform association with HLA-DQ alleles does not explain the clinical heterogeneity, especially the wide range in the age of onset of CD. We asked whether the age of onset of CD is also influenced by class I genes of the human MHC. We performed HLA typing in three groups of patients suffering from CD. The age of onset in the first group (N = 200) was before 15 years of age, in the second group (N = 62) between 15 and 40 years, in the third group (N = 59) after 40 years. We observed a statistically significant increase in the frequencies of HLA-B8 and Cw7 with increasing age of onset. In conclusion, we conclude that distinct alleles from the class I region of the human MHC might lead to late onset of CD. In particular, relatives of CD patients with the disease-prone HLA class I alleles HLA-B8 and Cw7 should be followed up carefully for late onset of CD. PMID- 12757180 TI - Immunolocalization and expression of kallistatin and tissue kallikrein in human inflammatory bowel disease. AB - The distribution of tissue kallikrein (TK) and its plasma inhibitor, kallistatin in plasma and intestinal tissue, was studied in patients with active ulcerative colitis (UC) and Crohn's disease (CD). TK was localized to goblet cells and kallistatin to epithelial cells of normal human intestine. Both proteins are visualized in macrophages inside granulomas in CD as well as in plasmocytes in both CD and UC. Intestinal tissue kallikrein (ITK) and kallistatin are significantly decreased in inflamed intestine compared to noninflammatory controls. TK mRNA is significantly decreased in intestinal biopsy samples from active UC patients compared with inactive patients or controls. Immunoreactive TK is present in plasma in very low concentrations in patients and did not differ in normal subjects. Plasma kallistatin was significantly decreased in patients with active disease compared to normal controls. Our data suggest that release of TK during inflammation plays a role in inflammatory bowel disease. PMID- 12757182 TI - Future directions for the clinical laboratory scientist. PMID- 12757181 TI - Biliary tract disease: a rare manifestation of eosinophilic gastroenteritis. AB - Eosinophilic gastroenteritis (EGE) is a rare inflammatory disease characterized by diffuse or scattered eosinophilic infiltration of the digestive tract and usually by peripheral blood eosinophilia. The most common presenting symptoms of EGE are abdominal pain, vomiting and diarrhea, but clinical features depend on which layers or location of gastrointestinal tract are involved. Treatment with corticosteroids results in clinical and histological remission in most patients and surgery can be avoided if a correct diagnosis is made. Previous history of allergy is a key to diagnosing EGE, but peripheral eosinophilia may be absent in some patients under concomitant treatment with corticosteroids. Radiological and endoscopic findings are also nonspecific and diagnosis must always be histologically confirmed. The gastrointestinal involvement is patchy in distribution, so more than one panendoscopic examination is often necessary to establish the diagnosis, and surgical or CT-guided full-thickness biopsy is needed in patients with muscular or serosal involvement. It emphasises the importance of a high index of clinical suspicion, which mainly depends on knowledge of natural history of the disease. We report here a case of EGE associated with transmural eosinophilic cholecystocholangitis, in a patient who presented with dyspeptic symptoms and recurrent cholestasis responsive to corticoesteroids. To our knowledge, this patient represents the second case, in the English literature, in which corticoid-responsive cholangitis was associated to histologically proven eosinophilic cholecystitis and gastrointestinal involvement, suggesting that EGE must always be considered in the differential diagnosis of biliary tract disease in patients with eosinophilia and/or atopic diseases. PMID- 12757183 TI - Winter 2003 issue of Clinical Laboratory Science. PMID- 12757184 TI - CLIA regulations updated. PMID- 12757185 TI - Trans type genotype alpha thalassemia trait: a case study. AB - This is a case of hypochromic, microcytic red cells in a young adult Caucasian female. It illustrates the importance of performing iron studies to confirm suspected iron deficiency anemia (IDA). Thalassemia minor is often misdiagnosed as IDA and iron therapy may be needlessly administered. Moreover, the patient will be unaware of an inherited hematological disorder which may require genetic counseling. alpha-thalassemia patients with the --/alphaalpha (cis) genotype should be advised of the risk for producing offspring with Hemoglobin H disease (genotype --/alpha-). In this case, DNA analysis confirmed the diagnosis of a trans type gene deletion alpha-thalassemia trait. Ancestry on the maternal side is German and French. On the paternal side the ancestry is Dutch and Scandinavian. Additionally, there was no knowledge of any family history of anemia on either the maternal or paternal side of the family. This case reaffirms that Anglo-Saxon ancestry does not preclude the diagnosis of alpha-thalassemia. It also supports the findings of Wang that when laboratory findings are suggestive of alpha-thalassemia minor, a moderately decreased MCV, slightly elevated red cell count, and the absence of hemoglobin H inclusions is probably indicative of trans rather than cis type gene deletion alpha-thalassemia trait. PMID- 12757186 TI - Career alternative: clinical trials. PMID- 12757187 TI - HIPAA Privacy Rule: the debate continues. PMID- 12757188 TI - Method comparison studies for prostate specific antigen and unconjugated estriol immunoassays. AB - OBJECTIVE: Method comparison studies were performed in order to move a semi automated prostate specific antigen (PSA) immunoassay and a manual unconjugated estriol (uE3) immunoassay to an automated chemistry immunoassay analyzer. The results of the two method comparison studies are compared. DESIGN: Serum samples collected on patients with physician orders for PSA or uE3 were assayed by both methods. PSA samples were assayed on a Hybritech Tandem Photon ERA and on two Beckman Coulter Access instruments. UE3 samples were assayed by RIA and on two Beckman Coulter Access instruments. Linear regression analysis was performed on both sets of data and within-run precision and dilution studies were performed on the PSA Access method. SETTING: Clinical chemistry laboratory, West Virginia University Hospitals Inc, Morgantown WV. RESULTS: PSA linear regression analysis for the two methods (ERA and Access 1) were y = 1.0008x + 0.0393, r = 0.9976, SE = 0.1319, n = 37 and (ERA and Access 2), y = 1.0019x + 0.0486, r = 0.9964, SE = 0.1632, n = 37. Within-run precision studies for both Access instruments produced acceptable coefficient variations and dilution study results were in PSA reportable range. uE3 linear regression analysis for the two methods (RIA and Access 1) were y = 1.4105x - 0.3741, r = 0.8696, SE = 0.8330, n = 33 and (RIA and Access 2) were y = 1.315x - 0.2292, r = 0.8643, SE = 0.7964, n = 33. CONCLUSION: The results of the method comparison studies for PSA were acceptable and the automated PSA immunoassay method was adopted. The results of the uE3 comparison studies did not show good correlation; the automated method was not adopted. PMID- 12757189 TI - Validity of injecting drug users' self report of hepatitis A, B, and C. AB - OBJECTIVE: To test the validity of drug users self-reports of diseases associated with drug use, in this case hepatitis A, B, and C. DESIGN: Injecting drug users (n = 653) were recruited and asked whether they had been diagnosed previously with hepatitis A, B, and/or C. These self-report data were compared to total hepatitis A antibody, hepatitis B core antibody, and hepatitis C antibody seromarkers as a means of determining the validity of the self-reported information. SETTING: Anchorage, Alaska. PARTICIPANTS: Criteria for inclusion included being at least 18-years old; testing positive on urinalysis for cocaine metabolites, amphetamine, or morphine; having visible signs of injection (track marks). INTERVENTION: Serological testing for hepatitis A, B, and C. MAIN OUTCOME: Findings indicate high specificity, low sensitivity, and low kappa coefficients for all three self-report measures. RESULTS: Subgroup analyses revealed significant differences in sensitivity associated with previous substance abuse treatment experience for hepatitis B self-report and with gender for hepatitis C self-report. CONCLUSION: Given the low sensitivity, the validity of drug users, self-reported information on hepatitis should be considered with caution. PMID- 12757190 TI - Managing the bleeding patient. PMID- 12757191 TI - Laboratory management of the bleeding patient. PMID- 12757192 TI - Use of blood products and factor concentrates for coagulation therapy. AB - Therapy of coagulation disorders has evolved from early use of fresh whole blood and plasma, to sophisticated recombinant factor concentrates. Although current testing protocols and viral inactivation methods ensure that transfusion of components is safer than ever, the potential for new threats continually exists, e.g., West Nile virus. Effective therapy depends on treating the specific deficiency with the safest and most appropriate replacement product, in the proper dose. PMID- 12757193 TI - Treatment of single factor deficiencies: a case study approach. PMID- 12757194 TI - Methodology for characterizing heat removal mechanism in human skin during cryogen spray cooling. AB - Cryogen spray cooling (CSC) reduces epidermal damage during laser treatment of various dermatoses. The goal of this study was to determine the heat removal mechanism in skin and quantify the amount in response to CSC. Thermocouples were imbedded in four model substrates with a range of thermal diffusivities, greater than three orders of magnitude in difference, to measure the temperature profiles in response to CSC and sapphire contact cooling, which removes heat completely by conduction. An algorithm solving an inverse heat conduction problem was subsequently used to quantify the amount of heat removal from the substrates using the measured temperatures. The interface thermal conductance and internal temperatures within the substrates were computed by a finite difference algorithm that solved the heat conduction equation. Results verify a marked increase in heat removal and interface thermal conductance with increasing thermal diffusivity. By estimation from the model substrate results, heat removal and interface thermal conductance values for skin were obtained. Data demonstrate that during CSC, evaporation is the dominant heat transfer mechanism in materials with higher thermal diffusivities; however, conductive cooling dominates in substrates with lower thermal diffusivities such as skin. PMID- 12757196 TI - Three-dimensional reconstruction of thrombus formation during photochemically induced arterial and venous thrombosis. AB - We have selected a model of photochemically induced thrombosis in hamsters and mice in which thrombus formation is visualized via transillumination and quantified via image analysis. Applying a gray-compensation method, the images of developing thrombi were presently transformed and a three-dimensional (3D) reconstruction of thrombus evolution performed off line. To this end, a nondimensional Gray-compensated parameter Gc was calculated. The integrated Gc (IGc) correlated linearly (r = 0.973) with the amount of light transilluminated and previously quantified as arbitrary light units. Matching Gc for reconstructed occlusive arterial and venous thrombi with the inner diameters of the hamster carotid artery and femoral vein, enabled the further conversion of IGc to real thrombus volumes, up to 0.14 mm in the carotid artery. In addition to enabling a graphical three-dimensional reconstruction of experimental thrombosis, via image subtraction, the kinetics of thrombus growth were visualized. Thus, platelet mediated thrombus growth was found to occur randomly in small thrombi, but in larger thrombi, it occurred preferentially in its tailing vortex in areas of recirculating flow. The present study therefore confirms in vitro findings in an in vivo model. The 3D reconstruction and kinetics of thrombus growth may be helpful in the mechanistic and pharmacological study of experimental thrombosis. PMID- 12757195 TI - Integrin-mediated preadipocyte adhesion and migration on laminin-1. AB - Cell adhesion and migration are key events in many biological processes and depend on extracellular matrix proteins. Understanding these cellular events in adipogenesis is paramount to elucidating the biological mechanisms underlying preadipocyte-specific aspects of obesity, diabetes, and adipose tissue development for the design of pharmaceutical screening and tissue engineering strategies. We quantitatively investigated preadipocyte adhesion and migration on laminin-1 surfaces and identified candidate cognate preadipocyte receptors for laminin-1. In adhesion studies, we found that preadipocytes readily adhered to laminin-1 as compared with other extracellular matrix proteins. In addition, immunocytochemical analysis demonstrated that an array of integrin molecules was present on the surface of preadipocytes. Preadipocyte adhesion on laminin-1 was quantitatively assessed using a sedimentation adhesion assay, and results suggested that preadipocyte adhesion to laminin-1 was mediated by the alpha1beta1 integrin. In addition, digital time-lapse microscopy and quantitative cell tracking revealed that inhibition of the alpha1beta1 integrin resulted in abrogation of preadipocyte migration on laminin-1 surfaces. These results strongly support the hypothesis that preadipocyte adhesion to and migration on laminin-1 substrata are regulated, in part, by integrins. PMID- 12757197 TI - Effect of specimen size and aspect ratio on the tensile properties of porcine aortic valve tissues. AB - The measurement of mechanical properties of biological tissues is subject to artifacts such as natural variability and inconsistency in specimen preparation. As a result, data cannot be easily compared across laboratories. To test the effects of variable specimen dimensions, we systematically modified the size and aspect ratio (AR) of porcine aortic valve tissues and measured their stiffness and extensibility. We found that: (i) as the AR of circumferential specimens increased from 1:1 to 5:1, their stiffness increased by 36% (p < 0.001) and their extensibility decreased by 21% (p < 0.001); (ii) as the AR of radial specimens increased from 0.8:1 to 4:1, their stiffness increased by 36% (p < 0.001) and their extensibility decreased by 34% (p < 0.001); (iii) as the size of circumferential specimens was reduced from 128 to 32 mm2 at fixed AR (2:1), their stiffness decreased by 6 (p = 0.05), and their extensibility increased by 17% (p < 0.001); and (iv) as the size of radial specimens was reduced from 72 to 32 mm2 at fixed AR (2:1), their stiffness decreased by 7% (p = 0.03) and their extensibility increased by 16% (p = 0.005). Thus, as specimens of constant length became narrower, they became stiffer and less extensible, and as specimens of fixed aspect ratio became smaller, they became less stiff and more extensible. Statistical models of these trends were predictive and can thus be used to integrate materials test data across different laboratories. PMID- 12757198 TI - Homogeneity of cardiac contraction despite physiological asynchrony of depolarization: a model study. AB - The use of mathematical models combining wave propagation and wall mechanics may provide new insights in the interpretation of cardiac deformation toward various forms of cardiac pathology. In the present study we investigated whether combining accepted mechanisms on propagation of the depolarization wave, time variant mechanical properties of cardiac tissue after depolarization, and hemodynamic load of the left ventricle (LV) by the aortic impedance in a three dimensional finite element model results in a physiological pattern of cardiac contraction. We assumed that the delay between depolarization for all myocytes and the onset of crossbridge formation was constant. Two simulations were performed, one in which contraction was initiated according to the regular depolarization pattern (NORM simulation), and another in which contraction was initiated after synchronous depolarization (SYNC simulation). In the NORM simulation propagation of depolarization was physiological, but wall strain was unphysiologically inhomogeneous. When simulating LV mechanics with unphysiological synchronous depolarization (SYNC) myofiber strain was more homogeneous and more physiologic. Apparently, the assumption of a constant delay between depolarization and onset of crossbridge formation results in an unrealistic contraction pattern. The present finding may indicate that electromechanical delay times are heterogeneously distributed, such that a contraction in a normal heart is more synchronous than depolarization. PMID- 12757199 TI - Voronoi polyhedra analysis of optimized arterial tree models. AB - Topological and metric properties of Voronoi polyhedra (VP) generated by the distal end points of terminal segments in arterial tree models grown by the method of constrained constructive optimization (CCO) are analyzed with the aim to characterize the spatial distribution of their supply sites relative to randomly distributed points as a reference model. The distributions of the number Nf of Voronoi cell faces, cell volume V, surface area S, area A of individual cell faces, and asphericity parameter alpha of the CCO models are all significantly different from the ones of random points, whereas the distributions of V, S, and alpha are also significantly different among CCO models optimized for minimum intravascular volume and minimum segment length (p < 0.0001). The distributions of Nf, V, and S of the CCO models are reasonably well approximated by two-parameter gamma distributions. We study scaling of intravascular blood volume and arterial cross-sectional area with the volume of supplied tissue, the latter being represented by the VP of the respective terminal segments. We observe scaling exponents from 1.20 +/- 0.007 to 1.08 +/- 0.005 for intravascular blood volume and 0.77 +/- 0.01 for arterial cross-sectional area. Setting terminal flows proportional to the associated VP volumes during tree construction yields a relative dispersion of terminal flows of 37% and a coefficient of skewness of 1.12. PMID- 12757201 TI - A deformable finite element derived finite difference method for cardiac activation problems. AB - We present a finite element (FE) derived finite difference (FD) technique for solving cardiac activation problems over deforming geometries using a bidomain framework. The geometry of the solution domain is defined by a FE mesh and over these FEs a high resolution FD mesh is generated. The difference points are located at regular intervals in the normalized material space within each of the FEs. The bidomain equations are then transformed to the embedded FD mesh which provides a solution space that is both regular and orthogonal. The solution points move in physical space with any deformation of the solution domain, but the equations are set up in such a way that the solution is invariant as it is constructed in material space. The derivation of this new solution technique is presented along with a series of examples that demonstrate the accuracy of this bidomain framework. PMID- 12757200 TI - A single equivalent moving dipole model: an efficient approach for localizing sites of origin of ventricular electrical activation. AB - We propose a new method for guiding catheter ablation procedures to abolish sites of origin of arrhythmias. This method models both cardiac electrical activity and current pulses delivered from the tip of the ablation catheter with a single equivalent moving dipole (SEMD). The SEMD parameters are obtained from analysis of body surface potentials. In this paper we examine the feasibility of this method by evaluating the performance of an inverse algorithm we developed to localize the SEMD from the surface potentials. In computer simulations realistic levels of measurement noise led to uncertainties in SEMD location approximately 0.005 cm. Dipole orientation randomization contributed to increased uncertainty (0.04 cm) in SEMD location only when boundary effects were included. In ventricular pacing swine studies, we found that the SEMD model accurately accounted for electrocardiographic wave forms and that measurement noise led to an uncertainty of approximately 0.04 cm in the SEMD at 15 ms after the pacing spike. We have also found that the algorithm we developed to identify the SEMD parameters yielded positions for two spatially separated pacing sites that maintained their direction and were very close to their physical separation. These results suggest that the SEMD method may potentially be used to guide radio frequency ablation procedures. PMID- 12757202 TI - An in vitro uniaxial stretch model for axonal injury. AB - We have developed a unique uniaxial stretching device to study axonal injury and neural cell death resulting from brain tissue deformations common in traumatic head injuries. Using displacement control rather than force control, this device is capable of achieving strains >70% and strain rates up to 90 s(-1), well above those currently used for studying axonal injury. We have demonstrated that the deformation of the specimen was uniaxial, uniform and highly reproducible; the prespecified displacement profiles could be realized almost precisely; and adequate cell adhesion could be achieved readily. The entire device can fit into a biological safety cabinet to maintain sterility, and the specimens are convenient for cell culture. This device can be used to investigate a wide range of biomechanical issues involved in diffuse axonal injury. PMID- 12757204 TI - Dynamics of the human head-neck system in the horizontal plane: joint properties with respect to a static torque. AB - The vestibular system has often been studied by perturbing the position of the head. This study was conducted to identify the dynamic properties of the head neck system in response to horizontal plane perturbations. A quasilinear approach was used to quantify the dynamics of the head-neck system at different levels of static torque. An operating point was established by applying a static torque to the head with a helmet-based perturber. The head-neck dynamics were then probed with a rich spectrum, stochastic, torque perturbation. Impulse response functions (IRFs) were estimated from correlation measures, and parametric models were fit to the IRFs. The results indicated that when the mean torque was held constant, the head-neck system behaved like a second-order, underdamped, passive system between 0.5 and 10.0 Hz. The system was not strictly linear, however. The properties of the system were sensitive to the static component of the torque. As the mean torque increased, the effective stiffness and damping progressively increased, and did so such that the system's damping ratio remained essentially constant. The findings of the study will assist in designing stimuli that are well tolerated by subjects and can induce head motions that span the performance capabilities of the vestibular system. PMID- 12757203 TI - Effect of altered matrix proteins on quasilinear viscoelastic properties in transgenic mouse tail tendons. AB - Tendons have complex mechanical behaviors that are viscoelastic, nonlinear, and anisotropic. It is widely held that these behaviors are provided for by the tissue's composition and structure. However, little data are available to quantify such structure-function relationships. This study quantified tendon mechanical behaviors, including viscoelasticity and nonlinearity, for groups of mice that were genetically engineered for altered extracellular matrix proteins. Uniaxial tensile stress-relaxation experiments were performed on tail tendon fascicles from the following groups: eight week old decorin knockout, eight week old reduced type I collagen, three week old control, and eight week old control. Data were fit using Fung's quasilinear viscoelastic model, where the model parameters represent the linear viscoelastic and nonlinear elastic response. The viscoelastic properties demonstrated a larger and faster stress relaxation for the decorin knockout and a smaller and slower stress relaxation for the three week control. The elastic parameter, A, in the eight week control group was significantly greater than in the collagen reduction and three week control groups. This study provides quantitative evidence for structure-function relationships in tendon, including the role of proteoglycan in viscoelasticity. Future studies should directly correlate composition and structure with tendon mechanics for the design and evaluation of tissue-engineered constructs or tendon repairs. PMID- 12757205 TI - An advanced approach for computer modeling and prototyping of the human tooth. AB - This paper presents a systematic and practical method for constructing accurate computer and physical models that can be employed for the study of human tooth mechanics. The proposed method starts with a histological section preparation of a human tooth. Through tracing outlines of the tooth on the sections, discrete points are obtained and are employed to construct B-spline curves that represent the exterior contours and dentino-enamel junction (DEJ) of the tooth using a least square curve fitting technique. The surface skinning technique is then employed to quilt the B-spline curves to create a smooth boundary and DEJ of the tooth using B-spline surfaces. These surfaces are respectively imported into SolidWorks via its application protocol interface to create solid models. The solid models are then imported into Pro/MECHANICA Structure for finite element analysis (FEA). The major advantage of the proposed method is that it first generates smooth solid models, instead of finite element models in discretized form. As a result, a more advanced p-FEA can be employed for structural analysis, which usually provides superior results to traditional h-FEA. In addition, the solid model constructed is smooth and can be fabricated with various scales using the solid freeform fabrication technology. This method is especially useful in supporting bioengineering applications, where the shape of the object is usually complicated. A human maxillary second molar is presented to illustrate and demonstrate the proposed method. Note that both the solid and p-FEA models of the molar are presented. However, comparison between p- and h-FEA models is out of the scope of the paper. PMID- 12757206 TI - Malaria genomics and disease control. PMID- 12757207 TI - The clinical and epidemiological features of childhood malaria in a moderately endemic area of Sri Lanka. AB - This study describes some clinical and epidemiological features of childhood malaria in a moderately endemic area of southern Sri Lanka. Six hundred and sixty two children, who experienced 1,138 attacks of malaria, and 172 children, who experienced 202 attacks of acute non-malarial fever, were followed over a period of two years. Of the 1,138 malaria infections followed, 776 were due to P. vivax, 359 were due to P. falciparum, and 3 were mixed infections. The majority of children presented within the first three days of the onset of symptoms. Headache (96%), feeling cold (81%) and arthralgia (77%) were the commonest presenting symptoms. Two hundred and sixty-four children experienced more than one attack of malaria. The clinical and epidemiological features of childhood malaria that have important implications for the planning and targeting of preventive measures are discussed. PMID- 12757208 TI - Use of routinely collected past surveillance data in identifying and mapping high risk areas in a malaria endemic area of Sri Lanka. AB - Stratification of malaria endemic areas on eco-epidemiological criteria is an important step in planning and implementing malaria control programs. The uses of stratification of malaria endemic areas lead to better targeting of control measures such as residual insecticide spraying in countries where unstable malaria transmission occur. In this study, two methods that can be used for stratification of malaria endemic areas in Sri Lanka using routinely collected surveillance data over a period of 9 years are described. In the first method, the median Annual Parasite Incidence (API) was used as the criterion to classify an area as at risk for malaria while in the second method, the API and the Falciparum Rate (FR) were used as the criteria. Risk maps were produced by plotting the results of the analyses on maps generated by EPIMAP. The potential uses of risk maps are discussed. PMID- 12757209 TI - Proguanil plus sulfamethoxazole in the treatment of uncomplicated Plasmodium falciparum malaria. AB - In a malaria endemic area of Brazil where P. falciparum is highly resistant to chloroquine and Fansidar, we conducted an in vivo study to evaluate the therapeutic response of proguanil plus sulfametoxazole against Plasmodium falciparum malaria. Twenty-five adult subjects with uncomplicated P. falciparum malaria received supervised drug administration and were followed for 28 days in an inpatient hospital or in a malaria free-transmission area. The therapeutic regimen was proguanil 100 mg BID plus sulfamethoxazole 1,000 mg BID for 7 days. Of those who took all medications (n=21), 17 (81%) were cured. Recrudescent parasitemia during follow-up occurred in four (19%) patients on days 14, 19, 20 and 21 after beginning of treatment. The remaining four (16%) subjects did not complete their therapeutic regimen because the incidence of side effects. Considering the shortage of falciparum malaria therapeutic options and the urgent need for new regimens to deal with the spread of drug resistant P. falciparum, one might consider the study results as a lead to study analogous compounds, hopefully with fewer adverse reactions. PMID- 12757210 TI - Plasmodium malariae in East Timor. AB - A community-based rainy-season malaria prevalence survey was conducted in Bobonaro district, in recently independent East Timor, in 2001. Although the survey was primarily aimed at defining the prevalence of Plasmodium falciparum and P. vivax, six individuals with P. malariae infection were identified (prevalence 0.57%). We believe these are the first reported cases of P. malariae from the island of Timor. PMID- 12757211 TI - Trypsin and aminopeptidase activities in blood-fed females Anopheles dirus (Diptera: Culicidae) of differing susceptibility to Plasmodium yoelii nigeriensis. AB - Midgut proteolytic enzymes contribute to the success or failure of Plasmodium infection of the mosquito. The present study investigated trypsin and aminopeptidase activities in the midgut of two strains of Anopheles dirus selected for susceptibility and refractoriness to Plasmodium yoelii nigeriensis. At intervals of 6 hours following a bloodmeal, the midguts of fully engorged female mosquitos were dissected, homogenized, and assayed for enzyme activity. No differences trypsin activity (nmole/min) were observed between the two strains throughout the course of blood digestion. By contrast, the aminopeptidase activity measured at 0 to 18 hours post-feeding was the same for the two strains, but at 24, 30 and 36 hours significantly less activity was observed in the refractory females. The results suggest neither trypsin nor aminopeptidase plays a role in the limitation of parasite development. PMID- 12757212 TI - Biting behavior and seasonal variation in the abundance of Anopheles minimus species A and C in Thailand. AB - We measured the seasonal abundance and bloodfeeding behavior of species A and C of the mosquito Anopheles minimus Theobald 1901 in an endemic malarious area of western Thailand. An. minimus s.l. is a major vector of human malarial and filarial parasites in Southeast Asia. Mosquitos were collected once a month for one year using four collection methods: human-baiting indoors, human-baiting outdoors, human-baiting in the forest, and cow-baiting. We found that both species A and C tend to feed from cows rather than humans; we did not find any preference for indoor, outdoor or forest-biting in either species. Both species had a peak biting density in October/November, at the end of the rainy season, and species C showed a second, smaller peak at the end of the cool season. These findings are discussed in relation to previous reports of the behavior of An. minimus s.l., particularly in light of suggestions that An. minimus s.l. has changed its feeding behavior in response to DDT spraying. PMID- 12757213 TI - Relative abundance of Culex quinquefasciatus (Diptera: Culicidae) with reference to infection and infectivity rate from the rural and urban areas of East and West Godavari districts of Andhra Pradesh, India. AB - This paper describes the seasonal abundance of Culex quinquefasciatus in the rural and urban areas of the East and West Godavari districts (EGDT and WGDT) of Andhra Pradesh, India. The per man-hour density (PMHD) was collected from seven units in EGDT and two units in WGDT, which comprised rural and urban areas. The highest infection and infectivity rates were found in the rural areas of Rajahmundry (43.6%) and Amalapuram (13.2%) respectively. In urban areas, the highest infection and infectivity rates were found in Rajahmundry: 7.5% and 3.6% respectively. There was considerable difference in the infection rate and infectivity rates between the rural areas and urban areas in each unit. PMID- 12757214 TI - Single-dose therapy for giardiasis in school-age children. AB - A randomized controlled trial was carried out to study the efficacy of combined albendazole and praziquantel in the treatment of giardiasis in school-age children. Eighty-four children were randomly allocated to 3 groups: group 1 (n = 31) albendazole 400 mg combined with praziquantel 20 mg/kg; group 2 (n = 26) albendazole 800 mg as a single dose; group 3 (n = 27) tinidazole 50 mg/kg as a single dose. The treatment was considered curative when Giardia was not found in two consecutive stool samples. The parasitological cure rate was 74.2% for combined single-dose albendazole-praziquantel, 50% and 92.6% in the albendazole and tinidazole groups respectively (p = 0.0023). There was no statistically significant difference between the cure rates of the combined regimen and tinidazole (p > 0.05). This combined regimen was considered safe, with only minor side-effects being observed. Of the single-dose regimens, tinidazole still achieves the highest parasitological cure rate for giardiasis. The albendazole praziquantel combined regimen may be an alternative single-dose therapy for giardiasis in children, especially as this combination will eradicate common intestinal protozoa and co-existing helminths. Whether the dosage of this combination treatment should be adjusted for G. intestinalis remains to be established by further study. PMID- 12757215 TI - Prevalence of Sarcocystis infection in horses in Mongolia. AB - Sarcocystis infection was detected in 93% of horses in Mongolia. Using the compress method, sarcocysts were found in the muscles of the diaphragm, heart and tongue in 40 of the 43 horses that were slaughtered at the Makh Impex Meat Company in Ulaan Baatar in July 1998. The muscle of the tongue showed the highest rate (97.5%) of infection. The distribution of sarcocysts in the muscles was positively correlated with horse age; the rate of detection was significantly lower (p=0.01) in the under 10 year old group than the older group. All horses were apparently healthy and were slaughtered for human consumption. PMID- 12757216 TI - Schistosoma japonicum strains: differentiation by RAPD and SSR-PCR. AB - Eight geographical isolates of Schistosoma japonicum from Taiwan and mainland China and one isolate of Schistosoma mansoni were studied by RAPD analysis using six arbitrary primers and SSR-PCR analysis using a (CA)8RY primer. The genetic distance was determined by the percentage of unshared bands. The RAPD and SSR-PCR results showed that the genetic distance between S. mansoni and S. japonicum was more than 0.900 and 0.850 respectively; the genetic distance between the eight geographical isolates of S. japonicum was 0.000 to 0.232 and 0.066 to 0.368 respectively. These results demonstrated the usefulness of RAPD and SSR-PCR for showing the differences of inter- and intra-species of Schistosoma. The results also suggest that there is genetic diversity among the different geographical strains of S. japonicum in China. PMID- 12757219 TI - Diversity of mollusks in the Lam Ta Khong reservoir, Nakhon Ratchasima, Thailand. AB - Sampling surveys to study the diversity of mollusks in the Lam Ta Khong reservoir, Nakhon Ratchasima Province, northeast Thailand, were carried out in the hot, rainy and cold seasons. The study area was divided into: Area I--the upper part where the Lam Ta Khong river drains; Area II--the mid-section of the reservoir; Area III--behind the dam. Mollusks were collected from four locations on each bank (to the right and left) of each area. Each location was sampled to include 6 cross-sectional stations; in total, 144 stations were sampled. In the deep water, an Ekman dredge was used to collect samples; the scoop or manual method was used at the water's edge. Ten species of snails and four species of clams were found. The dominant species of snails were: Clea helena, Bithynia siamensis goniomphalos and Melanoides tuberculata; clams were dominated by Corbicula sp. The intermediate host of the human blood fluke was not found. The population of most mollusk species increased during the cold season while that of clams and that of some species of snails increased during the rainy season. Clams and operculate snails predominated in Areas II and III. Pulmonate snails were mostly found close to the bank and on aquatic plants especially in Areas I and II. Operculate snails and clams mainly inhabited water 1 to 10 m deep. Two species of edible mollusks were found: Filopaludina martensi martensi and large numbers of Corbicula. Neither shedding light nor digestion with pepsin A revealed any human parasites in the mollusks sampled. PMID- 12757218 TI - The endoscopic diagnosis of intestinal capillariasis in a child: a case report. AB - A 13-year-old boy was diagnosed as having intestinal capillariasis by gastroduodenoscopy. He presented with a 10-month history of chronic abdominal pain and diarrhea. The boy had stayed in central Thailand and had eaten uncooked fish and raw shellfish. Gastroduodenoscopy showed normal jejunal mucosa although histology revealed flattened villi, crypt proliferation, acute inflammation, and eosinophilic granulomata. An egg of Capillaria philippinensis was also seen. The child was treated with mebendazole for 30 days. He had gained six kilograms by the time of his last follow-up. PMID- 12757220 TI - Seroprevalence of melioidosis in dairy cattle in Chiang Mai Province, northern Thailand. AB - The seroprevalence of melioidosis in dairy cattle in Chiang Mai Province was investigated using of the indirect hemagglutination antibody (IHA) method. Two hundred and fifty-three samples were tested for serum antibodies to Burkholderia pseudomallei. The samples were from a total population of 8,688 dairy cattle in the province; random sampling, stratified by the location of cattle, was used. The seroprevalence was determined as 2% at 1:40 cut-off value, which was estimated to equate to 0.3% to 3.7% (95% CI). This report of relatively low disease prevalence in the animal population corresponds to other prevalence studies of the agent in the environment and the human population in the region. The prevalence is markedly different to that reported from northeastern Thailand, where the disease is highly endemic. PMID- 12757217 TI - Intestinal parasites in children with diarrhea in Delhi, India. AB - The parasitic causes of diarrhea in children in Delhi were determined by the direct smear technique; stool specimens of 127 children were examined for intestinal parasites. In 59 cases (46.5%) intestinal helminths and protozoa were demonstrated. Ascaris lumbricoides was observed in 1 (0.8%) case, while Trichuris trichiura was the finding in 3 (2.4%). Protozoal parasites included Giardia intestinalis and Entamoeba histolytica in 14 (11%) cases each, Balantidium coli in 3 (2.4%) cases and Cryptosporidium spp in 24 (18.9%) patients. Mixed infection was not seen in any of the cases. Intestinal parasites may increase susceptibility to infection with other intestinal pathogens and therefore with the help of a simple technique, like direct fecal smear examination. rapid diagnosis can be made and specific therapy instituted. PMID- 12757221 TI - Antibody response in typhoid fever in endemic Indonesia and the relevance of serology and culture to diagnosis. AB - Culture and serology were performed on blood and serum samples collected at or shortly after admission from 473 patients presented with suspected clinical typhoid. Clinical symptoms at first presentation including confusion, hepatomegaly, splenomegaly, abdominal pain, anemia, and gastrointestinal bleeding were non-specific as they were observed even more often in non-typhoid patients. Culture confirmed the diagnosis in 65.3% of the patients with typhoid fever as the final diagnosis. The sensitivity (58%) and specificity (98.1%) of a rapid dipstick assay for the detection of S. typhi-specific immunoglobulin M were somewhat lower than those of culture but higher than those of the Widal test. The dipstick assay thus may well be used in the serodiagnosis of typhoid in situation where culture facilities are not available. Combination of test results of dipstick and culture improved sensitivity to 82.5%. In laboratories that perform blood culture the dipstick assay may be used as a rapid screening tests to facilitate a rapid diagnosis. Sensitivity of the dipstick assay strongly increased with duration of illness and was higher for culture positive than for culture negative patients. Duration of illness, and different pathogen and host factors including dose of infection, pathogenicity and antigenicity, and prior antibiotic use are likely to influence the immune response, therefore the result of the dipstick assay. Duration of illness and presence of S. typhi in the blood are major factors that determine severity of disease. PMID- 12757222 TI - Bacterial enteric pathogens in children with acute dysentery in Thailand: increasing importance of quinolone-resistant Campylobacter. AB - Current data on pathogen prevalence and drug resistance patterns are important for treatment and vaccine-development strategies. An etiologic study of acute bacterial dysentery was conducted in children up to 12 years of age in 2 major hospitals in and around Bangkok. Stool samples or rectal swabs and clinical data were collected. Standard microbiological methods were used to detect Salmonella, Shigella, Campylobacter, Vibrio, Aeromonas and Plesiomonas. Pathogenic E. coli (ETEC, EIEC, STEC) was identified by digoxigenin-labeled probes. A total of 623 cases were enrolled: median age 11.0 months (range 1 month-12 years). At least one bacterial pathogen was isolated in 55% of cases. Campylobacter was the most common pathogen found (28%), whereas Salmonella, Shigella and ETEC were isolated from 18%, 9% and 6% respectively. EIEC, Vibrio and Plesiomonas were isolated from <1% and no STEC was detected. C. jejuni serotypes 36, 4 and 11 were the most common. The mean age of cases with Campylobacter was significantly lower than with Shigella (17.9 vs 52.8 months, p<0.001). Clinical presentations of Campylobacter and Shigella infections were compared: fever (28% vs 37%), abdominal colic (62% vs 80%, p<0.05), vomiting (38% vs 70%, p<0.001) and bloody stools (52% vs 48%). The Campylobacter isolates (80% C. jejuni, 20% C. coli) were 90% resistant to ciprofloxacin but sensitive to macrolides. All the Shigella isolates (70% S. sonnei) were sensitive to quinolones. Our study illustrates the increasing importance of quinolone-resistant Campylobacter and the decline of Shigella in the etiology of dysentery in Thailand. The clinical presentation of campylobacteriosis is similar to that of shigellosis, except that the patients may be younger and there may be less association with colic and vomiting; having fecal leukocytes will be >10/HPF. The use of macrolide antibiotics rather than quinolones would be reasonable in children <24 months of age; fluoroquinolones will be ineffective in at least half of culture-positive cases. PMID- 12757223 TI - Evaluation of different primers for detecting mecA gene by PCR in comparison with phenotypic methods for discrimination of methicillin-resistant Staphylococcus aureus. AB - Detection of the mecA gene by polymerase chain reaction (PCR) is the gold standard for identifying methicillin-resistant Staphylococcus aureus (MRSA). PCR assays, employing MR1-MR2 primers (primer set 1) and MR3-MR4 primers (primer set 2) to generate 154 and 533 bp fragment, respectively, are most widely used for amplification of mecA gene. The purpose of this study was to evaluate the presence of mecA gene in 100 clinical isolates of S. aureus using PCR with the two pairs of primers. The results were compared to the broth dilution MIC method, oxacillin salt screening method (OSS) and oxacillin disk agar diffusion method (ODD). Fifteen of the 100 isolates showed a discrepancy between the mecA primer sets 1 and 2. Three isolates (3%) without the mecA gene showed discrepancies with phenotypic methods. The sensitivity, specificity and positive and negative predictive values for the 154 and 533 bp products of mecA were 79, 85, 83, 81 and 94, 100, 100, 94%, respectively. The results indicated that primer set 2 was more appropriate than primer set 1 for the detection of mecA gene in MRSA. There was a good correlation among the mecA gene detection, ODD and OSS methods. The discrepancy of three isolates between PCR and phenotypic methods should be clarified for other resistant mechanisms. PMID- 12757224 TI - Blood culture isolates from children admitted to Medical Unit III, Yangon Children's Hospital, 1998. AB - A one year study (August 1998-July 1999) of bacteremia in febrile children was carried out in the Medical Unit (III), Yangon Children's Hospital (YCH), Myanmar, to determine: (1) the bacteria responsible for fever of five days or more in children; (2) the antibiotic sensitivity pattern of these bacteria. Children aged one month to 12 years who had fever for five days or more and who did not receive antibiotics within the first 48 hours irrespective of the diagnosis were included in this study. A total of 120 patients fulfilled the criteria. Bacteria could be isolated from 65 cases (54.2%). The commonest organism isolated was Salmonella typhi (43.1%). Others included Escherichia coli (12.3%), Staphylococcus aureus (7.7%), Pseudomonas aeruginosa (7.7%); Streptococcus, Shigella, Diplococcus, Klebsiella and Acinetobacter were also isolated. The Salmonella typhi were resistant to conventional antibiotics (ampicillin, amoxicillin, chloramphenicol, and co-trimoxazole); however, they were sensitive to amikacin, netilmicin, nalidixic acid, and cephalothin. A cluster of enteric fever cases from Mingalartaungnyunt township was noticed and was reported to the Directorate of Health. Changes in the incidence and etiology of bacteremia in hospitals are well documented. Sentinel surveys of bacteremia in major hospitals should be carried out in order to detect the changing patterns of bacteremia and antibiotic sensitivity; such surveys will be of great help in establishing local antibiotic policies. PMID- 12757225 TI - Isolation and PCR detection of rickettsiae from clinical and rodent samples in Malaysia. AB - Isolation of rickettsiae from patients' blood samples and organ samples of wild rodents from areas with high seroprevalence of rickettsial infections was attempted using cell culture assay and animal passages. L929 mouse fibroblast cells grown in 24 well tissue culture plate were inoculated with buffy coat of febrile patients and examined for the growth of rickettsiae by Giemsa, Gimenez staining and direct immunofluorescence assay. No rickettsiae were isolated from 48 patients' blood samples. No symptomatic infections were noted in mice or guinea pigs infected with 50 organ samples of wild rodents. There was no rickettsial DNA amplified from these samples using various PCR detection systems for Orientia tsutsugamushi, typhus and spotted fever group rickettsiae. PMID- 12757226 TI - Septic shock secondary to scrub typhus: characteristics and complications. AB - Scrub typhus is an acute febrile illness caused by infection with Orientia tsutsugamushi transmitted by the bite of larval trombiculid mites (chiggers). A prospective study was conducted in septic shock patients in Maharat Hospital, Nakhon Ratchasima Province, Thailand, from 12 November 2001 to 5 January 2002. Of the 51 septic shock patients studied during the 7 week period, 18 (35.3%) were found to have evidence of scrub typhus infection; 3 patients (16.7%) died. In this study, septic shock caused by Orientia tsutsugamushi is the most prominent (35.3%) in endemic area of scrub typhus. Scrub typhus with septic shock patients results in organ failure: respiratory failure, DIC were predominant, followed by renal and hepatic involvement. Two deaths were due to respiratory failure and one death was as a result of combined respiratory and renal failure. Fever was the most common symptom, followed by headache, myalgia and dyspnea; lymphadenophathy and eschar are common signs. Laboratory findings revealed that almost all of the patients had a mild leukocytosis, reduced hematocrit and thrombocytopenia; SGOT, ALP, direct bilirubin (DB), total billirubin (TB), BUN, Cr were elevated; hypoalbuminemia was noted. Urinalysis showed that 88.9% of the patients had albuminuria. 77.8% of patients had abnormal chest X-rays. PMID- 12757227 TI - Borrelia burgdorferi (strain B. afzelii) antibodies among Malaysian blood donors and patients. AB - In this study, the presence of IgG and IgM antibodies against Borrelia burgdorferi (strain B. afzelii) among Malaysian blood donors and patients admitted to hospital with various infectious diseases was determined. Sera were screened using enzyme-linked immunosorbent assay (ELISA); positive sera were then subjected to Western blot testing. All but one of the blood donors were negative for borrelial antibodies. Of 121 patients' sera, IgM antibodies were detected in 24 (19.8%) and IgG antibodies were detected in 5 (4.1%) sera. Only one of two patients with skin manifestations suggestive of Lyme disease had IgM antibody against B. afzelii. Of 30 patients with exposure to tick typhus, 4 (13.3%) were IgM positive and 1 (3.3%) was IgG positive. Based on the detection of antigenic bands by Western blot, 6 patients' sera showed positive reactions. Antigenic bands of p39, p41 and p59/62 kDa were the commonest findings of Western blotting. This study provides serological evidence of B. afzelii infections in Malaysia; further investigation is needed to correlate serological and clinical findings. PMID- 12757229 TI - Prevalence of West Nile virus infection in India. AB - During the course of the virological investigation of cases of suspected viral fevers carried out at the National Institute of Virology (NIV), Pune, India, evidence of recent infection with West Nile (WN) virus was detected in 88 cases. Fever, general aches, headache, nausea and vomiting were the principal clinical features in 92% (81/88) of the cases; there were seven cases of encephalitis, in which WN virus-specific IgM class antibodies were detected in CSF samples. These cases of encephalitis were from Japanese encephalitis (JE) nonendemic areas, like Maharashtra and Rajasthan, as well as from JE endemic areas, like Goa and Orissa. Interestingly, neutralizing antibodies predominantly to WN virus were detected in CSF samples by the 50% cytopathic effect inhibition method; the titers ranged from 5 to 375. Cases of WN virus infection associated with both encephalitis and classic features have been reported for the first time in recent years in India. Reports of unique urban West Nile virus encephalitis epidemics in New York, Romania, and Algeria in recent years have signaled the emergence of neurological infection due to West Nile virus as a novel public health threat. This study is important because it records evidence of WN virus infection in India. PMID- 12757228 TI - A serological survey of arboviral diseases among the human population of the Andaman and Nicobar Islands, India. AB - In an attempt to determine the prevalence of certain arthropod-borne viruses of public health importance amongst the human population of the Andaman and Nicobar Islands of India, 2,401 sera were collected from six major localities. The sera were analysed by the hemagglutination inhibition (HI) and neutralization (N) tests, using Chikungunya (CHIK), Japanese encephalitis (JE), West Nile (WN), dengue (DEN-2), Langat (TP-21) and Kyasanur Forest disease (KFD) viral antigens. The highest prevalence of HI antibodies was detected against KFD virus (22.4%), followed by Langat (20.2%), JE (5.9%), DEN-2 (3.1%), CHIK (2.9%) and WN (0.8%) viruses. Cross-reactions to the viral antigens were also noted. The results of N tests indicated a high prevalence of DEN-2 (25.4%) virus, followed by Langat (17.5%), CHIK (15.3%), KFD (12%), JE (2.19%) and WN (1.8%). These results are discussed in relation to important epidemiological parameters like age, sex and geographical location. To our knowledge, this is the first report of an extensive serosurvey of arthropod-borne viruses on these islands. PMID- 12757230 TI - Risk factors for hypoxemia and respiratory failure in respiratory syncytial virus bronchiolitis. AB - Respiratory syncytial virus (RSV) bronchiolitis is a common infection in young children and may result in hospitalization. We examined the incidence of, and risk factors associated with, hypoxemia and respiratory failure in 216 children aged < 24 months admitted consecutively for proven RSV bronchiolitis. Hypoxemia was defined as SpO2 < 90% in room air and severe RSV bronchiolitis requiring intubation and ventilation was categorized as respiratory failure. Corrected age at admission was used for premature children (gestation < 37 weeks). Hypoxemia was suffered by 31 (14.3%) children. It was more likely to occur in children who were Malay (OR 2.56, 95%CI 1.05-6.23, p=0.03) or premature (OR 6.72, 95%CI 2.69 16.78, p<0.01). Hypoxemia was also more likely to develop in children with failure to thrive (OR 2.96, 95%CI 1.28-6.82, p<0.01). The seven (3.2%) children who were both premature (OR 11.94, 95%CI 2.50-56.99, p<0.01) and failure to thrive (OR 6.41, 95%CI 1.37-29.87, p=0.02) were more likely to develop respiratory failure. Prematurity was the only significant risk factor for hypoxemia and respiratory failure by logistic regression analysis (OR 1.17, 95%CI 1.06-1.55, p<0.01 and OR 1.14 95%CI 1.02-2.07, p=0.02 respectively). Prematurity was the single most important risk factor for both hypoxemia and respiratory failure in RSV bronchiolitis. PMID- 12757231 TI - Congenital hearing impairment associated with rubella: lessons from Bangladesh. AB - Infection with rubella virus during pregnancy may cause fetal death or the multiple congenital fetal abnormalities that are known as congenital rubella syndrome (CRS). Studies have demonstrated that congenital hearing impairment is the most frequent abnormality associated with intrauterine rubella infection. In the present study, the first of its kind in Bangladesh, we investigated the presence of rubella antibody in hearing-impaired children in order to understand the possible role of rubella infection in the development of hearing impairment. A total of 198 hearing-impaired children and 200 children without hearing problems were studied. After taking a detailed history from the parents, blood samples were collected from both mothers and children; sera were subjected to enzyme-linked immunosorbent assay (ELISA) for anti-rubella IgG. Rubella antibody was detected in 74% of the hearing-impaired children and in 18% of those with normal hearing: this finding correlated with the presence of rubella antibody in the mothers (67%) of rubella seropositive hearing-impaired children. In contrast, we observed rubella antibody in only 14% of the mothers of the children without hearing problems. Consistent with the presence of antibody, 41% of the seropositive mothers who had hearing-impaired children gave a history of fever and rash during early pregnancy. Our study indicates a strong association between rubella infection and hearing impairment in Bangladeshi children. In addition, it also indicates that infection by rubella virus is common in Bangladesh: this suggests that priority should be given to implementing appropriate measures for the control of rubella. PMID- 12757233 TI - The spectrum of HIV-related disease in rural Central Thailand. AB - To determine the spectrum of HIV-related illnesses presenting to a rural primary and secondary healthcare facility in Central Thailand, a cross-sectional study was conducted. Routinely collected data were extracted from outpatient medical notes for all adult HIV-infected new attenders of the Manorom Christian Hospital Infectious Disease Clinic. Data concerning inpatient admissions of HIV-infected individuals were collected from ward admission books and discharge summaries. Complete data were available for 229 outpatients, 70% of whom were men. The median age at presentation was 31 years for men and 30 years for women. The majority of the outpatients were married (69%) and infected heterosexually (91%). The commonest conditions requiring admission were cryptococcal meningitis (15%), bacterial pneumonia (12%), extrapulmonary tuberculosis (12%), Pneumocystis carinii pneumonia (7%), cerebral toxoplasmosis (4%) and pulmonary tuberculosis (3%). Of the patients presenting for the first time, 32% had AIDS-defining illnesses. Presentations with some conditions, such as tuberculosis and septicemia, were fewer than expected. The common opportunistic infections among HIV-infected adults in this rural region are treatable and preventable. Most patients present with advanced disease and earlier diagnosis, through improved access to voluntary counseling and testing, would enable them to receive appropriate preventive therapies and antiretrovirals as they becomes available. The high prevalence of cryptococcal disease suggests that prophylactic anti fungal therapy may be of value in this area. Septicemia and tuberculosis may be under-diagnosed, highlighting the need for improved diagnostic laboratory facilities or treatment based upon validated clinical algorithms. Community care and palliative care services for HIV-infected individuals will increasingly be required in this region. PMID- 12757232 TI - Prevalence of HIV-1 drug resistance in antiretroviral-naive pregnant Thai women. AB - HIV-1 drug resistance may limit the use of antiretrovirals when attempting to reduce the vertical transmission rate. Establishing the prevalence of the HIV-1 mutations associated with antiretroviral resistance in pregnant women will enable clinicians to maximize the chances of preventing vertical transmission. In order to determine the prevalence of HIV-1 resistant strains among antiretroviral-naive pregnant Thai women, the nucleotide sequences of the HIV-1 polymerase (pol) gene were evaluated. The plasma samples were collected from the women during the 34th week of pregnancy: numerous secondary mutations could be found in the reverse transcriptase (RT) and protease gene, while no primary mutations in the pol gene were found. The result also showed that by detecting the delta32bp deletion within the CCR 5 locus, it was evident that none of HIV-1 infected individuals had homozygous or heterozygous delta32bp deletions of the CCR5 gene; moreover, no CCR5 gene mutations were found in any individual. PMID- 12757234 TI - Assessment of current epidemiological status of viral hepatitis in Guangdong Province, China. AB - Descriptive epidemiology was employed to analyze the current epidemiological status of viral hepatitis and strategies for control of viral hepatitis in Guangdong Province, China. The average incidence rate (IR) was 52.21 per 100,000 during 1991-2001 with a decrease of 37.98% from 79.10 per 100,000 in 1991 to 44.61 per 100,000 in 2001. Hepatitis B constituted 79.1% of total hepatitis cases. The Pearl River Delta Region had the highest prevalence of viral hepatitis, with the highest IR in the 20-35 age-group and the highest seasonal IR from March to May. Since 1991 HAV cases have declined gradually but HBV cases rose gradually with an increase of 2.48 times in constitutive ratio and with an increase of 53.35% in IR from 1991 to 2001. HBV vaccination in the newborn resulted in decreasing IR of the viral hepatitis and HBsAg carrier rate in infants. As to an increase of IR of the viral hepatitis in the 20-35 age group, it is suggested that HBV vaccination should be performed in adults and juveniles, especially in the social workers and the employees in public services. PMID- 12757236 TI - Etiology of endemic viral hepatitis in urban North India. AB - This study was carried out to determine the presence of markers of hepatitis viruses in patients with acute liver disease. Coinfection of HAV, HBV, HCV, and HEV was studied. Sera from 306 patients with a clinical diagnosis of acute liver disease were tested for the presence of anti-HAV antibody, HBsAg, anti-HBc antibody, anti-HBs antibody, anti-HCV antibody and IgM anti-HEV antibody by ELISA. Liver function tests were correlated with the presence of infection. Of the 306 cases, 7 (2.3%) had IgM anti-HAV, 9 (2.9%) had IgM anti-HBc, 37 (12.1%) had HBsAg, 84 (27.4%) had anti-HBs, 10 (3.3%) were HCV infected and 63 (20.6%) had IgM anti-HEV. There was no significant difference in the clinical and liver function profiles of infected and uninfected patients. Similarly, no difference was observed in cases coinfected with more than one virus compared with those infected with a single pathogen. HEV had the highest prevalence amongst our cases. There was no difference in the clinical profiles of patients with non-A, non-B, non-C, non-E hepatitis by antibody assays and testing for viremia could be helpful in making the correct diagnosis. PMID- 12757235 TI - The prevalence of viral hepatitis among the Hmong people of northern Thailand. AB - Sera from 269 Hmong people (102 males and 167 females, with mean age 35.4 years, range 16-63 years) were examined in order to determine the seroprevalence of hepatitis virus infection. The seroprevalence rates for HAV (hepatitis A virus), HBV (hepatitis B virus), HCV (hepatitis C virus), HDV (hepatitis D virus), HEV (hepatitis E virus), HGV (hepatitis G virus) and TTV (TT virus) infection were 87.8% (n=140), 76.0% (n=150), 2.0% (n=150), 0.7% (n=150), 6.5% (n=139), 5.3% (n=94) and 25.6% (n=121) respectively. The rate for carriers of HBV (HBsAg) was 13.8% (20.5% in males and 9.6% females) with a peak prevalence in the 21-40 year age group. A high rate of HAV seropositivity was found among the younger subjects. The rate of HEV seroprevalence was low. The prevalence of TTV-DNA was high with no difference between the sexes. HGV-RNA prevalence was low and seen primarily in males. This study indicates that the Hmong people are endemically infected with HAV and HBV infection and should be considered for targeted vaccination. The role of TTV and HGV in producing illness and hepatic disease has yet to be determined in this population. PMID- 12757238 TI - Levels of serum vascular cell adhesion molecule-1 in measles. AB - While the levels of serum vascular cell adhesion molecule-1 (VCAM-1) have been investigated in many diseases, they have not, to our knowledge, been studied in patients with measles. Serum VCAM-1 and creatinine levels were determined in six adolescent and adult Japanese patients with measles in the acute febrile phase and defervescent afebrile phase. Serum VCAM-1 levels were significantly higher in the acute febrile phase than in the convalescent afebrile phase, but no significant difference between the serum creatinine levels was shown in the two phases. Our study revealed that the high serum VCAM-1 level in the acute febrile phase later decreased in the afebrile convalescent phase. The measurement of circulating VCAM-1 may be useful for the assessment of convalescence in patients with measles. PMID- 12757237 TI - Prevalence and genotypes of hepatitis C virus infection among drug addicts and blood donors in Thailand. AB - Hepatitis C virus (HCV) is an infectious agent that has the potential to cause chronic liver disease, cirrhosis and hepatocellular carcinoma. We determined the prevalence and genotypes of HCV infection among groups of drug addicts: intravenous drug users (n = 134), methamphetamine users (n = 100), inhaled-drugs users (n = 19) and alcoholics (n = 50); a group of blood donors acted as a control. The control group consisted of 179 randomly-selected anti-HCV positive samples: these were subjected to HCV RNA screening and genotyping. The anti-HCV test was performed by ELISA: HCV RNA screening was by nested RT-PCR that employed primers from the 5' noncoding region. The genotype assay was based upon analysis of the 5' NCR amplified sequences and RFLP. Hepatitis C virus was highly prevalent among all groups of drug addicts (12-70%). In 2000. among the new blood donors (n = 66,340) at the National Blood Center, Thai Red Cross, anti-HCV prevalence amounted to 0.98%. The HCV genotype distribution showed that the most prevalent genotype was 3a, followed by 1b and 6a. Our data demonstrated the very high prevalence of HCV infection in IVDUs, a finding that is consistent with the blood-borne nature of the virus. In order to curb HCV infection, a determined effort to educate both the general population and high-risk groups is required; such a program of education would address both general and particular methods of transmission, especially the use of non-sterile needles etc. PMID- 12757239 TI - Hemoglobinopathies among five major ethnic groups in Karachi, Pakistan. AB - A brief survey of abnormal hemoglobin variants among the major ethnic groups of Karachi was conducted; 202,600 subjects were studied. Patients with low hemoglobin (Hb), low mean cell volume (MCV) and mean cell hemoglobin (MCH) including anemia, microcytosis, hypochromic hemolysis and target cells, were refered for the identification of hemoglobinopathy by molecular methods. Population screening showed that 60% had iron-deficiency anemia and 40% had hemolytic anemia, of which 20.6% was due to beta-thalassemia major, 13% beta thalassemia trait, 5.1% sickle cell disease, 0.76% hemoglobin D Punjab (HbD Punjab), 0.32% hemoglobin C (HbC), and 0.22% hereditary persistence of fetal hemoglobin (HPFH). PMID- 12757240 TI - The validation of an automated liquid chromatography system for the diagnosis of thalassemias and hemoglobinopathies. AB - Accurate and precise hemoglobin separation and the quantitation of Hb A2 and Hb F are essential for the diagnosis of the thalassemias and hemoglobinopathies. Presented in this study is the validation of the the Hb A2 assay of the HbGold analyzer, a fully automated liquid chromatography system for hemoglobin separation and quantitation. Variability of Hb A2 quantitation was quite low; the CV's of within-run, between-run and interlaboratory studies were 1.8-3.1%, 3.4 6.0% and 6.8-8.8% respectively. The results of the %Hb A2 quantitated by HbGold analyzer correlated well with those given by the Bio-Rad Variant Hb testing system (r=0.98). The application of the HbGold analyzer for the diagnosis of the thalassemia phenotypes frequently observed in Thailand is considered. In conclusion, the Hb A2 assay of the HbGold analyzer could be used for the quantitation of Hb A2 and Hb F and the presumptive identification of abnormal hemoglobins. PMID- 12757241 TI - Factor V Leiden and prothrombin G20210A mutations in Thai patients awaiting kidney transplant. AB - Renal transplantation provides the best long-term treatment for chronic renal failure, but thrombosis of the transplanted renal artery or renal vein is one of the causes of kidney failure in the early postoperative period. Factor V Leiden (FVL) and prothrombin G20210A mutation are the most frequent genetic abnormalities associated with venous thrombosis. We investigated the prevalence of FVL and prothrombin G20210A by polymerase chain reaction with restriction fragment length polymorphism in 75 Thai patients awaiting renal transplant, and a control group of 106 healthy blood donors. Of those awaiting renal transplant, none was found to carry FVL or prothrombin G20210A mutations. Neither the heterozygous nor the homozygous FVL mutation nor the prothrombin G20210A mutation was detected in the 106 healthy volunteers. Although we failed to detect FVL and prothrombin G20210A mutation among those waiting for a kidney transplant, the population size was small. Further studies need to be performed in order to ascertain if these coagulation mutations are of relevance in predicting patients at risk of early transplant failure. PMID- 12757242 TI - Hematologic abnormalities in recurrent oral ulceration. AB - The aim of this study was to analyse the hematologic status in patients with recurrent oral ulceration (ROU). Twenty-three patients with ROU and 19 control subjects were examined consecutively for hematological abnormalities including serum folate, red cell folate and vitamin B12 levels. Their complete blood counts, hemoglobin typing, serum and red cell folate and serum vitamin B12 levels were studied. Low red cell folate levels were found in 11 out of 23 patients (47.83%) with ROU. They were defined as having folate deficiency (n=5), folate deficient erythropoiesis (n=1), and folate depletion (n=5). The serum and red cell folate levels in the control group were within normal range. There was a statistically significant low red cell folate in the ROU compared to the control group (p=0.000). The serum vitamin B12 levels were within normal range in both ROU and control groups. Hemoglobin, hematocrit and mean corpuscular hemoglobin concentrations were in the normal range in both groups and none had anemia or macrocytosis. PMID- 12757243 TI - High urine ttMA levels among fishermen from a Thai rural village. AB - The ongoing industrialization of Thailand, a developing country in Southeast Asia, has put many occupations at high risk of benzene exposure. However, there are few reports about monitoring the biomarkers of benzene exposure among Thais. In this study, we report on high urine trans, trans-muconic acid (ttMA) levels among the fishermen of a rural community. Using high-performance liquid chromatography (HPLC) for urine ttMA determination, 49 subjects (30 fishermen and 19 control subjects) were studied. The mean urine ttMA level in fisherman (0.180 +/- 0.130 mg/g creatinine) was significantly higher than that of the control group (0.015 +/- 0.053 mg/g creatinine) (p < 0.05). We recommend the monitoring of urine ttMA in these workers. The monitoring of the possible benzene contamination of the water and fish is recommended for further study. PMID- 12757244 TI - Infectious threats from exotic pets: dermatological implications. AB - Zoonoses are diseases that can be transmitted from animals to humans. More than 250 distinct zoonoses have been described in the literature. It is estimated that 56% of United States households contain at least one pet, and although considerable research has been performed regarding the more common household animals including dogs, cats, small birds, and rodents, surprisingly little is known about the zoonotic hazards of owning the more exotic pets. According to the 1997 USPHS/IDSA Report on the Prevention of Opportunistic Infections in Persons Infected with Human Immunodeficiency Virus, the immunocompromised patient should avoid contact with feces-laden soil, litter boxes, reptiles, most pet birds, and any animal less than 6 months old . It has also been documented that because of their inquisitive nature, children are at even higher risk for infection from animals than adolescents or immunocompetent adults. In this article the authors have reviewed the available data regarding hazards associated with the hedgehog, flying squirrel, iguana, chinchilla, and cockatoo. With the growing popularity of such exotic pets, further observation and research is warranted. Physicians need to be aware of the possibility of zoonotic disease related to exotic pet ownership, and they should address this issue when obtaining a history and formulating a differential diagnosis of cutaneous lesions suggestive of such illnesses. PMID- 12757245 TI - Tick-borne infections. AB - Ticks, obligate, blood-sucking members of the order Acarina and class Arachnida, are the most common agents of vector-borne diseases in the United States. Ticks play an important role in transmitting viruses, bacteria, spirochetes, parasites, and rickettsia. This article reviews the epidemiology, microbiology, diagnosis, and treatment of the major tick-borne diseases in the United States. PMID- 12757246 TI - Infections with Vibrio vulnificus. AB - V. vulnificus is an uncommon cause of soft tissue infection and primary septicemia, especially in patients with hepatic disease or who patients who are immunocompromised. The mortality of infection in these patients is extremely high despite timely antibiotic therapy. It is important to consider the possibility of infection with V. vulnificus in patients who present with high fever and rapidly progressive sepsis and have a history of consumption of raw seafood (especially oysters) or exposure of open wounds in a marine environment. Public education regarding the risk of raw seafood consumption is essential to preventing infection with this virulent pathogen. PMID- 12757247 TI - Protothecosis. AB - Intravenous amphotericin B remains the most effective drug for eradicating Prototheca infections. It should be used as a first-line agent in cases of disseminated disease and in patients with severe underlying illness or with immunosuppression or immunocompromise. Azole antifungals and surgery should be reserved for patients with more localized disease. Itraconazole appears to be the most effective agent of this drug class, and it should be administered at 200 mg/day for 2 months. Surgical excision should be considered as a first-line therapy in patients who present with olecranon bursal infections. PMID- 12757248 TI - Non-dermatophyte onychomycosis. AB - Non-dermatophyte organisms are becoming increasingly prevalent in onychomycosis. This apparent emergence might be an artifact of improved diagnostic techniques and increased awareness that these fungi are potential etiologic agents. It is important to bear in mind that all isolated organisms should be evaluated as potential pathogens when diagnosing fungal infections, especially given the increasing use of immunosuppressive drugs and the increasing numbers of chronically immunocompromised individuals. While many patients with non dermatophyte mold onychomycosis will respond to oral or topical antifungal therapy, poor or incomplete response might still be expected in some patients. PMID- 12757249 TI - New and emerging pediatric infections. AB - Several aspects of emerging pediatric cutaneous infections are of importance to the clinician. New manifestations of parvovirus infection should be recognized promptly, especially because transmission to susceptible populations might lead to serious complications. In the immunocompromised pediatric population, the outcome of cutaneous mold infections can be improved with prompt recognition and initiation of treatment. The incidence of community-acquired MRSA infections in pediatrics is becoming more than a regional concern, and this coincides with the issue of limited antibiotic susceptibility for MRSA as well as other infections. New antibiotics such as linezolid are emerging as potential treatments for drug resistant pathogens. An older group of antibiotics, the fluoroquinolones, appear to be well tolerated in children and should not be withheld from this population when the benefits of treatment out-weigh the risks. PMID- 12757250 TI - Ivermectin. AB - Ivermectin is a potent antiparasitic drug and the first commercially available member of a new class of drugs (macrocyclic lactones) that has been approved for human use. Ivermectin has already proven to be highly effective in the elimination of river blindness as a public health burden. Side effects have been minor, and patient acceptance is good. Promising results in off-label applications for ectoparasitic infestations are increasingly important as resistance to topical therapy becomes more prevalent Ivermectin represents an advance in the therapeutic armamentarium and should be considered in appropriate cases. PMID- 12757251 TI - Albendazole: a new drug for human parasitoses. AB - In 1961 Brown and his team discovered that thiobendazoles were highly effective against gastrointestinal nematodes. This discovery led to the development of albendazole, the most recent of the benzimidazolic drugs. Albendazole is used against numerous animal and human parasites and it is the authors' first choice as drug treatment of cutaneous larva migrans. PMID- 12757252 TI - Imiquimod. AB - Imiquimod is the first of the immune response modifiers to stimulate a localized immune response to treat infectious skin conditions. The reported TLR-7 activation to provoke an immune response suggests that imiquimod might mimic a microbial antigen. The immune response initiated by induced production of IFN alpha and TFN-alpha is specifically aimed at an infectious antigen and appears mediated (in part) by enhanced migration of Langerhans' cells to regional lymph nodes. The approved indication for imiquimod is for treatment of genital warts. The drug has demonstrated a 50% to 60% clearance rate and a 12% to 20% recurrence rate for this indication (Table 1). This recurrence rate is the lowest reported among the currently recommended treatment modalities. The self-applied treatment avoids costly and painful office-based procedures. Case reports and open-label studies have demonstrated the efficacy of imiquimod in treating some cases of common, plantar, and flat warts, as well as molluscum contagiosum and leishmaniasis. Common and plantar warts respond better to imiquimod in combination with cryosurgery, occlusion, and keratolytics. Reports of successful imiquimod treatment of granuloma annulare, alopecia areata, and vitiligo might suggest an infectious etiology to those conditions, although this hypothesis is highly speculative. PMID- 12757253 TI - Topical cidofovir for the treatment of dermatologic conditions: verruca, condyloma, intraepithelial neoplasia, herpes simplex and its potential use in smallpox. AB - Cidofovir is a new antiviral drug that has a broad spectrum of activity against several DNA viruses. Many of the disorders caused by these viruses do not have satisfactory therapy, and given the efficacy of this agent in treating many of these conditions, it holds great promise. It is hoped that ongoing studies will confirm the initial anecdotal reports regarding its therapeutic efficacy and lack of systemic side effects. It is also hoped that the cost to formulate and use cidofovir topically will eventually decrease to a level that will allow more widespread use of this drug. PMID- 12757254 TI - Management of acyclovir-resistant herpes simplex virus. AB - In immunocompetent patients, HSV is controlled rapidly by the human host's immune system, and recurrent lesions are small and short lived. When treated with antiviral agents, these patients rarely develop resistance to these drugs. In contrast immunocompromised patients might not be able to control HSV infection. Thus, frequent and severe reactivations are often seen and might lead to fatal herpetic encephalitis or disseminated HSV infection. Treatment in these patients is limited because immunocompromised hosts often develop severe herpes disease refractory to antiviral drug therapy. It is therefore imperative that physicians develop regimens to deal with both receptive and refractory HSV disease. The following treatment protocol (modified from Balfour and colleagues) might serve as a guide until further investigation of new drugs is performed. In all patients standard oral ACV therapy should be initiated at a dose of 200 mg orally, five times a day for the first 3 to 5 days. Prior to treatment, cultures the lesions should be obtained to verify HSV etiology. If the response is poor, the dose of oral ACV should be increased to 800 mg five times a day. If no response seen after 5 to 7 days, it is unlikely that the lesion will respond to intravenous ACV (or chemically and structurally related drugs such as VCV or famciclovir), so an alternative regimen must be assigned. First, repeat cultures for vital, fungal, and bacterial pathogens must be performed. In addition, ACV susceptibility studies should be ordered, if available. If the mucocutaneous lesion is accessible for topical treatment, TFT (as ophthalmic solution) should be applied to the area three to four times a day until the lesion is completely healed. If the lesion is inaccessible or if the response to TFT is poor, therapy with intravenous foscarnet should be given for 10 days or until complete resolution of the lesions. The dosage of foscarnet should be 40 milligrams per kilogram three times per day or 60 milligrams per kilogram twice daily. If foscarnet fails to achieve clinical clearing, consideration should be given to use of intravenous cidofovir (or application of compounded 1% to 3% topical cidofovir ointment). Vidarabine is reserved for situations in which all of these therapies fail. If lesions reoccur in the same location following clearing, the patient should started on high-dose oral ACV (800 mg, five times daily) or intravenous foscarnet (40 mg/kg tid or 60 mg/kg bid) as soon as possible. When lesions occur in a different location, the patient should be treated initially with standard doses of oral ACV (200 mg, five times daily) and the above protocol should be followed should there be clinical failure. In the future, new treatment options for patients with documented HSV resistance will be important in reducing the clinical impact of HSV. PMID- 12757255 TI - Unusual infectious complications of dermatologic procedures. AB - Because dermatologic procedures disrupt skin integrity, they alter the body's protective barrier and predispose individuals to cutaneous infection. Postoperative wound infections--even with common pathogens such as S. aureus- seldom complicate dermatologic procedures; however, unusual infections have been reported to complicate excisions, biopsies, skin grafts, chemical peels, dermabrasion, laser resurfacing, liposuction, blepharoplasty, and injections (eg, with anesthetic solutions or botulinum toxin). Numerous environmental and patient risk factors increase the rate of postoperative wound infections, but otherwise healthy individuals undergoing relatively simple procedures are sometimes affected. Obtaining a thorough patient, history (including history of prior HSV infection or any immunocompromising factors) is crucial. Patients should be warned of potential complications, particularly when they are undergoing cosmetic procedures. It is important to maintain a high index of suspicion for possible wound infection in all patients that extends several months postoperatively. Manifestations of unusual postoperative infections are highly variable, and they might be secondary to bacterial, fungal, viral, or parasitic pathogens. Bacterial lesions are often polymicrobial, and bacterial superinfection can exacerbate other wound complications such as HSV reactivation. Most wound infections remain localized, but occasionally systemic disease occurs. For example, cutaneous diphtheria or rapidly growing mycobacteria rarely disseminate, whereas TSS results in systemic disease caused by toxin release. Some unusual postsurgical infections are self-limited, but they can still be potentially life threatening or disfiguring. Antimicrobial prophylaxis might reduce the risk of wound infection in some cases. Clinicians can better care for patients by becoming familiar with the causes and clinical manifestations of unusual dermatologic postoperative wound infections (Table 1). Following the recognition of an infectious process, appropriate diagnostic procedures allow for pathogen identification and the prompt institution of indicated therapy. PMID- 12757256 TI - Antibiotic use in dermatologic surgery. AB - Few situations in dermatologic surgery require prophylactic antibiotics. The AHA has decreased the dose for endocarditis prophylaxis from antibiotics before and after the procedure to only 1 hour prior to the procedure. In the 1997 guidelines, fewer procedures are listed as requiring antibiotics compared with prior guidelines. In fact, several authors have questioned the efficacy of prophylactic antibiotics. The sequela of endocarditis or an infected prosthetic joint are certainly serious and possibly life-threatening conditions, yet this should not be a justification for using a therapy that is not proven and has potential serious side effects of its own. The authors suggest not using antibiotics on clean or clean-contaminated wounds regardless of cardiac history. Patients with prosthetic joint replacements probably do not need prophylactic antibiotics in cutaneous surgery unless mucosa is invaded; in such cases the guidelines set by the ADA and the AAOS should be followed. The authors believe that antibiotics should be reserved for contaminated or infected wounds when their application is therapeutic. Table 2 contains a summary of the authors' recommendations for the use of antibiotics in cutaneous surgery. Each patient should be evaluated on an individual basis, and consultation with the patient's primary physician, cardiologist, or orthopedist should be sought when the need arises. PMID- 12757257 TI - Vaccines for viral diseases with dermatologic manifestations. AB - Vaccines against infectious diseases have been available since the 1800s, when an immunization strategy against smallpox developed by Jenner gained wide acceptance. Until recently, the only vaccination strategies available involved the use of protein-based, whole killed, and attenuated live virus vaccines. These strategies have led to the development of effective vaccines against a variety of diseases with primary or prominent cutaneous manifestations. Effective and safe vaccines now used worldwide include those directed against measles and rubella (now commonly used together with a mumps vaccine as the trivalent MMR), chickenpox, and hepatitis B. The eradication of naturally occurring smallpox remains one of the greatest successes in the history of modern medicine, but stockpiles of live smallpox exist in the United States and Russia. Renewed interest in the smallpox vaccine reflects concerns about a possible bioterrorist threat using this virus. Yellow fever is a hemorrhagic virus endemic to tropical areas of South America and Africa. An effective vaccine for this virus has existed since 1937, and it is used widely in endemic areas of South America, and to a lesser extent in Africa. This vaccine is recommended once every 10 years for people who are traveling to endemic areas. Advances in immunology have led to a greater understanding of immune system function in viral diseases. Progress in genetics and molecular biology has allowed researchers to design vaccines with novel mechanisms of action (eg, DNA, vector, and VLP vaccines). Vaccines have also been designed to specifically target particular viral components, allowing for stimulation of various arms of the immune system as desired. Ongoing research shows promise in prophylactic and therapeutic vaccination for viral infections with cutaneous manifestations. Further studies are necessary before vaccines for HSV, HPV, and HIV become commercially available. PMID- 12757258 TI - Wound healing: a multidisciplinary approach for dermatologists. AB - This article has provided a review of common and some less common approaches to wound healing. Chronic wound healing is one of the more challenging areas of medicine, with a nice balance of the science and art of medicine. An evidence based, patient-centered approach can be used to effectively improve the care of many difficult to heal ulcers in often frustrated patients. The multidisciplinary wound clinic concept can work to improve the outcomes of patients with leg ulcers. PMID- 12757259 TI - Insights into the mitochondrial signaling pathway: what lessons for chemotherapy? AB - Mitochondria are potent integrators/coordinators of apoptosis signaling pathways. Indeed, under physiological conditions, the initiation of apoptosis leads to the accumulation of second messengers that converge on mitochondria. In response, these organelles undergo a membrane permeabilization, presumably due to the opening of protein channels, culminating in the release of proapoptotic proteins into the cytosol. Under pathological conditions, a failure of mitochondrial membrane permeabilization (MMP) can result in an inhibition of apoptosis and enhanced resistance to chemotherapy. Several non-mutually exclusive mechanisms may account for a defect in the execution or regulation of MMP. These include (i) alterations in gene transcription, (ii) gene mutations resulting in protein inactivation, and (iii) defects of intracellular localization. This may concern structural proteins of the permeability transition pore complex, as well as MMP regulatory proteins, such as Bax/Bcl-2 family members, p53, and cyclophilin D. Analysis of these mechanisms should improve our understanding of the basic function of mitochondria in apoptosis and help elaborate new strategies to correct MMP failure from a therapeutic perspective. PMID- 12757260 TI - Studies on the production of IL-15 in HIV-infected/AIDS patients. AB - IL-15 is essential for the development and differentiation of NK cells. It selectively induces proliferation of CD8+ memory T lymphocytes. Despite its importance in both innate and adaptive immune responses, little is known about its production in HIV-infected persons. We report here that IL-15 levels are significantly decreased in the sera of HIV-infected/AIDS patients compared to control sera. We also show that PBMC from the infected patients are compromised in their ability to respond with enhanced production of IL-15 upon exposure to HSV-1. The decreased production of IL-15 occurs despite a comparable increase in IL-15 mRNA in the PBMC of HIV-infected and healthy HIV-seronegative donors when exposed to HSV-1. The HSV-stimulated patients' PBMC exhibited less NK activity compared to similarly treated normal PBMC. These results suggest that a compromised ability of PBMC from HIV-infected individuals to induce IL-15 production in response to a viral stimulus may be a reason of their compromised innate and adaptive immunity. PMID- 12757261 TI - Lupus manifestations in severe combined immunodeficient (SCID) mice and in human/mouse radiation chimeras. AB - The objective of this study was to develop a human lupus model. To this end we have established and compared two models: (1) severe combined immunodeficient (SCID) mice reconstituted with peripheral blood lymphocytes (PBL) of either systemic lupus erythematosus (SLE) patients or healthy controls and (2) lethally irradiated BALB/c mice radioprotected with bone marrow of SCID mice, to which human PBL were transferred (human/mouse chimera). Engraftment was successful in most (78.4%) recipient mice as determined by the levels of human IgG measured. In about 50% of either SCID mice or human/mouse chimeras that were successfully engrafted with PBL of SLE patients, significant anti-dsDNA autoantibodies, mostly of the IgG1 and IgG2 isotypes, were determined. Interestingly, in a significant number (84.5%) of recipients of PBL of the healthy controls, anti-dsDNA antibodies were observed as well, suggesting that PBL of at least some of the healthy controls have the potential to develop SLE-associated autoantibodies under the appropriate stimulatory conditions. Glomerular immune deposits (human IgG, mouse C3) were detected in 70-80% of SCID mice with human DNA specific antibodies and in a third of the human/mouse chimeras. Thus, SLE serology and glomerular pathology were reproducibly demonstrated in two models of human SLE. These models should allow the evaluation of potential therapies for the treatment of lupus patients. PMID- 12757262 TI - Dehydroepiandrosterone therapy ameliorates experimental autoimmune myasthenia gravis in Lewis rats. AB - To detect a possible effect of dehydroepiandrosterone (DHEA) in the pathogenesis of experimental autoimmune myasthenia gravis (EAMG), DHEA (0.5 mg/rat) was administrated intraperitoneally to Lewis rats every other day from day 4 postimmunization (p.i.) to day 35 p.i. with Torpedo acetylcholine receptor (AChR) and Freund's complete adjuvant. Rats treated with DHEA had a lower clinical score (mean clinic score, 2 versus 0.5 on day 37 p.i.) and a lower body weight loss (mean body weight, 169 versus 142 g on day 37 p.i.) compared with control EAMG rats. DHEA treatment decreased serum anti-AChR IgG and IgG2b antibody titers on days 7, 14, and 21 p.i. and inhibited the levels of anti-AChR IgG antibody secreting cells (60%), accompanied by decreased IL-4 (33%) and augmented TGF beta1-positive cells (41%) among lymph node mononuclear cells. These results obtained from EAMG in Lewis rats further encourage us to study DHEA treatment in human MG. PMID- 12757263 TI - Transcript expression of two Iglambda rearrangements and RAG-1/RAG-2 in a mature human B cell producing IgMlambda islet cell autoantibody. AB - A human B cell clone, EBV-MB91, producing IgMlambda islet cell autoantibody (ICA), obtained by Epstein-Barr virus (EBV) transformation of peripheral CD5- surface Ig+ B cells from a Type 1 diabetic child, and an EBV-MB91-derived hetrohybridoma, HY-MB91, were analyzed for rearranged Ig genes. Both EBV-MB91 and HY-MB91 contained and expressed a unique IgH chain rearrangement (unmutated VH5 51-D6-19-JH5) but contained and expressed two Iglambda chain rearrangements: (i) Vlambda1-4-Jlambda3-Clambda3, which encoded the Iglambda chains (pI, 8.0) of IgMlambda-ICA, showing few mutations but consistent with Ag-driven selection according to the multinomial probability model; and (ii) Vlambda4-1-Jlambda3 Clambda3, with more mutations but inconsistent with antigen-driven selection and involving stop codons that precluded Iglambda synthesis. HY-MB91 showed a progressive loss of IgMlambda-ICA secretion, which was coupled with transcripts of the aberrant Vlambda4-1-Jlambda3-Clambda3 predominating (1.7-fold) over those of Vlambda1-4-Jlambda3-Clambda3. EBV-MB91 also showed the loss of IgMlambda-ICA secretion, associated with cell death. RAG-1 and RAG-2 transcripts occurred in EBV-MB91 but not in HY-MB91, indicating that the former but not the latter might have been able to exhibit V(D)J recombinase activity. Data show that a mature nonmalignant human B cell clone producing IgMlambda-ICA can express RAG-1/RAG-2 transcripts. That the aberrant Vlambda4-1-Jlambda3-Clambda3 was a nonproductive rearrangement occurring at the pre-B cell stage cannot be excluded. However, the hypothetical possibility that one of the two rearrangements corresponded to a secondary rearrangement occurring in the mature B cell represented by the EBV MB91 clone might also be considered and is discussed. PMID- 12757264 TI - IL-4 expression delays eosinophil-independent vasculopathy and fibrosis during allograft rejection in the mouse. AB - Transplant vasculopathy in the mouse is thought to be dependent on IL-4 and mediated by IL-5 and eosinophils, whereas in the rat and human systems, IL-4 is associated with the absence of transplant vasculopathy and down-regulation of a Th1-type response. In this study we tested the possibility that the apparent difference in the role of IL-4 in transplant vasculopathy is related to protocol differences rather than to the species being studied. Using a protocol that closely resembles that used in rat and human studies, we developed a model of transplant vasculopathy in the mouse that is associated with Th1-type cytokines and independent of IL-5 and eosinophil infiltration. In this model IL-4 promotes a significant delay in vasculopathy in the graft (P = 0.04) and a decrease in the incidence of allograft rejection (P = 0.02). The data suggest that the role of IL 4 in transplant vasculopathy can be controlled by the protocol used to treat the transplant recipient. PMID- 12757266 TI - Chronic active Epstein-Barr virus infection associated with low expression of leukocyte-associated immunoglobulin-like receptor-1 (LAIR-1) on natural killer cells. AB - Chronic active Epstein-Barr virus (CAEBV) infection is an uncommon and severe manifestation of EBV infection that frequently leads to the development of natural killer (NK) or T cell lymphomas. We report here that NK cells derived from a boy with CAEBV showed decreased cytotoxic function and impaired interferon gamma secretion against a variety of transformed target cell lines. In addition, NK cells from the patient lacked expression of the broadly expressed NK receptor, leukocyte-associated immunoglobulin-like receptor-1 (LAIR-1). These data suggest that CAEBV is associated with low expression of LAIR-1 on NK cells. PMID- 12757265 TI - Treatment with an anti-CD14 monoclonal antibody delays and inhibits lipopolysaccharide-induced gene expression in humans in vivo. AB - CD14 is a receptor important for activation of cells by lipopolysaccharide (LPS). Treatment with the CD14 antibody IC14 was previously found to attenuate the release of proinflammatory cytokines and some chemokines into the circulation of healthy humans intravenously injected with LPS. To determine the role of circulating leukocytes in CD14-dependent gene expression, 16 healthy volunteers received LPS preceded by either IC14 or placebo. At different time points, mRNA was isolated from whole blood and gene expression was determined by multiplex ligation-dependent probe amplification (MLPA). LPS induced MIP-1alpha, MIP-1beta, IL-8, IL-1beta, and IL-1Ra mRNA production, which was delayed by 1 hr and reduced twofold by IC14 treatment. TNFR1 was unresponsive, whereas other investigated cytokines remained undetectable. Further, LPS showed differential effects on NFkappaB gene expression. LPS induced IkappaBalpha production, whereas p50 was unresponsive and p65 and p49/p100 remained undetectable. LPS induced IkappaBalpha expression was delayed (1 hr) and reduced by IC14. Gene expression profiles in blood cells corresponded poorly with observed changes in plasma levels. These data suggest that peripheral blood cells are of negligible importance in LPS induced production of inflammatory mediators in vivo and that LPS may activate genes via a CD14-independent pathway that is slower and less efficient. PMID- 12757267 TI - Will reference pricing address the health cost conundrum? PMID- 12757268 TI - Reference pricing for drugs: is it compatible with U.S. health care? AB - To control spending on prescription drugs, health insurance systems abroad have experimented in recent years with a novel form of patient cost sharing called "reference pricing." Under this approach, the insurer covers only the prices of low-cost, benchmark drugs in therapeutic clusters that are deemed to be close substitutes for one another in treating specific illnesses. Patients who desire a higher-price substitute in a cluster must then pay the full difference between the retail price of that drug and the reference price covered by the insurer. This paper explores the difficult trade-offs that policymakers must make in designing such a system, drawing where relevant from experience abroad. PMID- 12757269 TI - Dilemmas in regulation of the market for pharmaceuticals. AB - What can be learned from international experience of efforts to control spending and to improve efficiency and access in pharmaceutical markets? Policymakers tend to reinvent many policies to control the behavior of patients, doctors, and industry, despite a lack of evidence of those policies' cost-effectiveness. There is an emerging consensus that reimbursement in public and private health care systems should be informed by evidence of the cost-effectiveness of treatments and that utilization should be constrained by budget caps and information systems. Whatever the policy chosen, evaluation is as essential as it is rare. PMID- 12757270 TI - The efficient use of pharmaceuticals: does Europe have any lessons for a Medicare drug benefit? AB - Managing drug use in a way that maximizes the value obtained from total health care spending faces obstacles; hence, payers and policymakers tend to look at pharmaceutical expenditures in isolation from the rest of health care spending. Currently there are both regulatory and putative market-based approaches to containing pharmaceutical spending worldwide. But evidence suggests that regulatory efforts in Europe and elsewhere have not proved effective in containing costs or improving efficiency or access, and supposedly market-based solutions now in vogue, such as reference pricing, pose their own set of challenges and may in practice violate market principles. In the end, silo-based budgeting is short-sighted; the emphasis in Europe and in the United States should be on measures that achieve efficient health care rather than the containment of drug spending. PMID- 12757271 TI - Government commitment and regulation of prescription drugs. AB - Two papers in this volume review efforts worldwide to control the growth of drug spending and discuss the potential role for the U.S. government with respect to rationing of prescription drugs. I put the roles given to government in the two papers in context by focusing on the role of government as a partner with the pharmaceutical industry. I concentrate on the unique features of the prescription drug market, coupled with the fact that government is a payer, regulator, and provider in the health sector. I conclude that the federal government should exercise caution when attempting to regulate prescription drug prices. PMID- 12757272 TI - Whither seniors' pharmacare: lessons from (and for) Canada. AB - Canada's provincial governments have, until relatively recently, provided virtually all seniors with generous prescription drug coverage. Managers of these programs have implemented a variety of policies to contain spending while ensuring access to necessary medicines. Some of these policies have been successful in temporarily slowing cost growth. However, the lack of comprehensive utilization management tools has resulted in ongoing spending increases that now constitute a threat to the sustainability of a public drug subsidy for Canadian seniors. Sustainable and equitable pharmacare programs require the political willingness to confront opposition to policies that will, if successful, contain program costs without obstructing access. PMID- 12757273 TI - Taiwan's new national health insurance program: genesis and experience so far. AB - In 1995, after a planning effort of about half a decade, the Republic of China (Taiwan) replaced a previous patchwork of separate social health insurance funds with one single-payer, national health insurance scheme that is administered by an agency of the central government's Department of Health. Within a year this bold legislative act brought the health care utilization rates of the 41 percent of Taiwan's hitherto uninsured population up to par with those of the previously insured population. This paper describes the achievements of this policy initiative so far, along with the growing pains it has encountered, and seeks to extract lessons from the experience for health policymakers in other countries. PMID- 12757274 TI - Does universal health insurance make health care unaffordable? Lessons from Taiwan. AB - This paper examines the performance of Taiwan's National Health Insurance (NHI), a universal health insurance program, implemented in 1995, that covers comprehensive services. The authors address two key questions: Did the NHI cause Taiwanese health spending to escalate to an "unaffordable" level? What are the benefits of the NHI? They find that Taiwan's single-payer NHI system enabled Taiwan to manage health spending inflation and that the resulting savings largely offset the incremental cost of covering the previously uninsured. Under the NHI, the Taiwanese have more equal access to health care, greater financial risk protection, and equity in health care financing. The NHI consistently receives a 70 percent public satisfaction rate. PMID- 12757275 TI - It's the prices, stupid: why the United States is so different from other countries. AB - This paper uses the latest data from the Organization for Economic Cooperation and Development (OECD) to compare the health systems of the thirty member countries in 2000. Total health spending--the distribution of public and private health spending in the OECD countries--is presented and discussed. U.S. public spending as a percentage of GDP (5.8 percent) is virtually identical to public spending in the United Kingdom, Italy, and Japan (5.9 percent each) and not much smaller than in Canada (6.5 percent). The paper also compares pharmaceutical spending, health system capacity, and use of medical services. The data show that the United States spends more on health care than any other country. However, on most measures of health services use, the United States is below the OECD median. These facts suggest that the difference in spending is caused mostly by higher prices for health care goods and services in the United States. PMID- 12757276 TI - Common concerns amid diverse systems: health care experiences in five countries. AB - This article reports on a comparative survey of sicker adults in Australia, Canada, New Zealand, the United Kingdom, and the United States. The study finds that despite differences among the health care systems, large proportions of citizens across the five countries report dissatisfaction with their health care system and serious problems including medical and medication errors, faulty patient-physician communication, and poor care coordination. The most crucial policy implication of these findings is that a focus on a small population of intensive health system users could have the potential to both control costs and improve care. PMID- 12757277 TI - The prevalence of treated and untreated mental disorders in five countries. AB - We analyzed survey data from Canada, Chile, Germany, The Netherlands, and the United States to study the prevalence and treatment of mental and substance abuse disorders. Total past-year prevalence estimates range between 17.0 percent (Chile) and 29.1 percent (U.S.). Many cases are mild. Although disorder severity is strongly related to treatment, one- to two-thirds of serious cases receive no treatment each year. Most treatment goes to minor and mild cases. Undertreatment of serious cases is most pronounced among young poorly educated males. Outreach is needed to reduce barriers to care among serious cases and young people at risk of serious disorders. PMID- 12757278 TI - Public reporting on quality in the United States and the United Kingdom. AB - The public reporting of comparative information about health care quality is becoming an accepted way of improving accountability and quality. Quality report cards have been prominent in the United States for more than a decade and are a central feature of British health system reform. In this paper we examine the common challenges and differences in implementation of the policy in the two countries. We use this information to explore some key questions relating to the content, target audience, and use of published information. We end by making specific recommendations for maximizing the effectiveness of public reporting. PMID- 12757279 TI - The impact of a national prescription drug formulary on prices, market share, and spending: lessons for Medicare? AB - Several recent bills in Congress to add a Medicare prescription drug benefit would allow the use of formularies to control costs. However, there is little empirical evidence of the impact of formularies among elderly and disabled populations. We assess the effect of a closed formulary implemented by the Veterans Health Administration (VHA) in 1997 on drug prices, market share, and drug spending. We find that the VHA National Formulary was effective at shifting prescribing behavior toward the selected drugs, achieving sizable price reductions from manufacturers, and greatly decreasing drug spending. PMID- 12757280 TI - Provider risk sharing in Medicaid managed care plans. AB - Provider risk sharing was common throughout the 1990s. Recent evidence suggests waning interest, although no information exists that is specific to Medicaid. This paper examines risk-sharing arrangements in Medicaid managed care through a survey of participating plans in eleven states conducted during 2001. Risk sharing is prevalent among Medicaid-participating plans and often involves traditional providers. The "flight from risk" that others describe is not yet apparent in Medicaid, but Medicaid's idiosyncrasies might mean that trends appearing in other lines of business do not apply. PMID- 12757281 TI - Wealth patterns among elderly Americans: implications for health care affordability. AB - This paper estimates the ability of the elderly to pay for necessary health care services and emerging technologies. Projections from the Long Term Care Financing Model paint a promising picture of the income and assets that elders in the future will have available to support discretionary, uncovered health care and service costs. Nevertheless, policymakers should pay close attention to the finances of the "Tweeners"--people who are middle class with low levels of discretionary assets available for health and long-term care. PMID- 12757282 TI - Do seniors get the medicines prescribed for them? Evidence from the 1996-1999 Medicare Current Beneficiary Survey. AB - Using the 1996-1999 Medicare Current Beneficiary Survey, we examine trends in nonacquisition of prescribed medicines among seniors. Each year less than 3 percent of seniors reported not getting the medicines that were prescribed for them, most often for economic reasons, but they also noted reasons relating to their preferences about using medicines. The presence or absence of drug coverage and chronic disease did not appreciably change this percentage, which suggests that a Medicare drug benefit may not greatly increase seniors' acquisition of their prescribed medicines. PMID- 12757283 TI - On being a grantmaker. PMID- 12757284 TI - From the talk to the walk. PMID- 12757285 TI - High out-of-pocket health care spending by the elderly. AB - We use data from the Health and Retirement Study to examine the elderly's out-of pocket health care spending. We find that Medicare HMOs, employer supplements, and Medicaid effectively insulate against the risk of high expenditures. At the ninetieth percentile, Medicare beneficiaries with employer supplements or enrolled in Medicare HMOs spend 1,600 dollars less out of pocket than beneficiaries with traditional Medicare spend. For the poor elderly, Medicaid offers similar protection. Among the near-poor elderly, there is little employer coverage, so Medicare HMOs provide most of the protection against financial risk. There is evidence that Medicare HMO benefits have eroded since 1998, raising the question of whether the near-poor have lost financial protection since then. PMID- 12757286 TI - Health insurance for workers who lose jobs: implications for various subsidy schemes. AB - A number of proposals have been made to help laid-off workers purchase health insurance. We use data from the 1996 Medical Expenditure Panel Survey to profile the insurance status of workers who left a job. Our descriptive analysis suggests that it might be difficult to design policies that target those who would otherwise be uninsured and that large subsidies might be needed to help laid-off workers. PMID- 12757288 TI - Toward a policy agenda on medical research funding: results of a symposium. AB - University of the Sciences in Philadelphia convened a symposium to discuss the roles of government, industry, and foundations in funding biomedical research. Government, no longer the largest funder of biomedical research, focuses on basic science. The pharmaceutical industry, now the largest sponsor, focuses on developing and testing new products. Foundations play a unique role in their ability to fund research overlooked by the other sectors. However, gaps remain in this infrastructure. Barriers, including lack of qualified investigators and administrative burdens, constrain discovery. Funders should collaborate to address these constraints and stimulate new sources of funding. PMID- 12757287 TI - Rehabilitation therapy in skilled nursing facilities: effects of Medicare's new prospective payment system. AB - In 1998 the Centers for Medicare and Medicaid Services (CMS) began phasing in a new prospective payment system (PPS) for Medicare payments to skilled nursing facilities (SNFs). I examine the effects of the new PPS on the level of rehabilitation therapy provided in SNFs. The percentage of residents of freestanding SNFs receiving extremely high levels of rehabilitation therapy dropped significantly, and the percentage receiving moderate levels increased. Freestanding SNFs, particularly for-profits, dramatically altered the services they provided in response to new financial incentives. This responsiveness underscores the importance of efforts now under way to refine the SNF PPS. PMID- 12757289 TI - Quality improvement efforts. PMID- 12757290 TI - Hospitals disclosing harm. PMID- 12757291 TI - Telling patients the truth. PMID- 12757292 TI - International medical graduates. PMID- 12757293 TI - Medicaid and the disabled. PMID- 12757294 TI - Medicaid and mental health. PMID- 12757295 TI - Medicaid recipients' rights. PMID- 12757296 TI - Medicaid and managed care. PMID- 12757297 TI - Preventing chronic disease. PMID- 12757298 TI - Looking ahead with anticipation and hope. PMID- 12757299 TI - ACEN response to health accord. PMID- 12757300 TI - Leadership in research: about building relationships. PMID- 12757301 TI - Leadership is present in the commonplace acts. PMID- 12757302 TI - What is leadership in nursing administration? PMID- 12757303 TI - Reaping what we sow: nursing education and leadership in Canada and the United States. PMID- 12757304 TI - Profile: Dorothy Wylie. PMID- 12757305 TI - Exploring an alternative metaphor for nursing: relinquishing military images and language. AB - The language used to describe nursing practice and nursing leadership has a profound influence on how nurses think about themselves, their work relationships, and indeed the very essence of their reason for being. Language often includes metaphor in order to help capture the complexities and layers of meaning that establish contexts for action. Nurses and others have relied on various metaphors to describe nursing work. However, there is one metaphor that, more than any other, has shaped the context of nursing work and formed the images and the meanings that nurses have of themselves and their purposes in practice. The privileged one is the military metaphor. This article explores the notion of metaphor, and its usefulness and potential to help nurses change their work patterns. The traditions and history of the military metaphor are examined and an alternative notion of the "frontier" is proposed in order to enhance understanding of the potential for change. PMID- 12757306 TI - Partnership in research: a tandem of opportunities and constraints. AB - Partnership is a term that is occurring more and more frequently in the research lexicon, an approach that is gradually becoming a sine qua non in the field of health and healthcare research in Canada. The purpose of this article is to share thoughts and experiences regarding research carried out in partnership. The relevance and necessity of partnerships in strategic health research will be examined, and the contribution of partnerships to the development and "re centring" of intra- and interdisciplinary knowledge and knowledge transfer will be discussed. Based on the nursing and related-fields literature, the key elements of partnership, and the advantages and disadvantages of this strategy to pursuing research projects will be presented. An important issue in a professional discipline such as nursing will be discussed, i.e. the intra disciplinary partnership between researchers and clinicians. Strategies that could enhance this particular type of partnership and avenues for catalyzing the synergy that must perforce develop, over the coming years, will be proposed. PMID- 12757307 TI - Pathfinding for nursing science in the 21st century: where to from here? AB - In late 2002, the Office of Nursing Policy convened a "Think Tank" of Canadian nurse scientists and policy-makers to create an opportunity to articulate the major challenges in advancing the nursing science agenda in Canada, and also to discuss what action(s) might be taken to address these challenges. Together, participants identified a number of challenges as follows: * increasing the pool and critical mass of nurse scientists * maximizing career trajectory * aligning focus of research with sources of research funding * enhancing linkages between practice and science * building research programs evaluating scientific productivity, and recognizing individual scientists * capitalizing on nursing research opportunities and * investing in leadership and succession planning. Participants subsequently selected four of these challenges as priorities and identified key collaborative opportunities to address them. Given this promising start, it seems that nursing is positioning itself to make a significant contribution to health research. PMID- 12757308 TI - Alternative work arrangements: senior administrative job-share. AB - As the healthcare workforce ages and workers expect increasingly flexible work environments, employers are looking for creative strategies to maintain a healthy and vital workforce. This paper offers one such strategy. It describes a job share experience by two senior administrators within a large healthcare authority and provides the findings from a survey evaluation. PMID- 12757309 TI - Cultivating APNs for the future: a hospital-based advanced practice nursing internship program. AB - This paper describes a clinical nurse specialist/clinical nurse specialist-nurse practitioner (CNS/CNS-NP) internship program in a tertiary care teaching hospital Hamilton Health Sciences, located in south-central Ontario, Canada. The goal of the program is to increase the number of fully qualified CNS/CNS-NPs in the organization. With this program, Hamilton Health Sciences (HHS) is able to recognize and reward nurses already employed within the hospital. The program assists nurses in transition to the CNS/CNS-NP role while they are completing the required educational requirements. The authors describe the components of the internship program, responsibilities of participants and the infrastructure required to support the program. Overall the internship program is a valuable mechanism for supporting the development of advanced practice roles in the hospital setting. PMID- 12757310 TI - Learning circles: collaborating to promote RN and LPN role enhancement. AB - Recent events in the Newfoundland and Labrador healthcare system, including changes to nursing scope of practice, have had a profound effect on practice environments for direct care registered nurses (RNs) and Licensed practical nurses (LPNs). This article describes an initiative introduced by the Association of Registered Nurses of Newfoundland and Labrador (ARNNL) and the Council for Licensed Practical Nurses (CLPN) in response to concerns raised by healthcare providers related to collaboration and teamwork between RNs and LPNs. Through the Learning Circles Project, the ARNNL and CLPN committed to fostering an improved working relationship for nursing team members based on the values of working together, mutual respect and an enhanced working environment. PMID- 12757311 TI - Healthcare restructuring with a view to equity and efficiency: reflections on unintended consequences. AB - This paper is developed from a research study that examined the hospitalization and helpseeking experiences of diverse ethnocultural populations in the era of healthcare restraint. Interview data were gathered from 60 patients while hospitalized and after their discharge home. Fifty-six healthcare professionals, the majority of whom were nurses caring for these patients while they were in hospital, were also interviewed. The data gathered in this study provides evidence to illustrate how restructuring associated with fiscal restraint designed to enhance efficiencies while ensuring the provision of medically necessary services, has had unintended consequences for some groups of patients and for nurses. These consequences have created a context for inequities in care delivery for those most vulnerable. In this paper we trace the ways in which the changed context of care delivery has exerted its effects on both nurses and patients and illustrate how each has sought to bridge gaps created when organizational supports are lacking. Our study data offer insight into the complexities of the practice setting and difficulties that arise when resources cannot be mobilized to match patients' needs. Our analysis examines how tensions between ideologies of efficiency and accessibility are navigated at the front lines, and draws attention to unintended consequences of the current policy context. PMID- 12757312 TI - How we do it: Acute pancreatitis. PMID- 12757313 TI - Case of the month. Solid pseudo-papillary tumor of the pancreas (SPT) and incidental mature cystic teratoma in the pelvis. PMID- 12757314 TI - Anterior mediastinal extension of primary chest wall infections: role of spiral CT in detection and management. AB - The purpose of this review was to define the role of spiral CT in the diagnosis and clinical management of patients with mediastinal infections resulting from extension of anterior chest wall infections. The review focuses on determining the frequency of mediastinal extension of chest wall infections as well as to define the patterns spread. SUBJECTS AND METHODS: CT scans of 22 consecutive patients collected over a 2-year period with anterior chest wall infections were evaluated both prospectively and retrospectively. Clinical, pathologic, and CT characteristics of the infections were evaluated. The extension of the infection into the mediastinum as well as the role of CT in diagnosis and management of these patients was examined. RESULTS: Of the 22 patients, 10/22 (45.4%) showed CT evidence of mediastinal infection. Extension of chest wall infection occurred in patients with both soft tissue and sternoclavicular joint processes. The assessment of mediastinal involvement was best seen at the level of the great vessels origin from the aortic arch. CT findings in the mediastinum included fat hyperdensity and stranding, fluid collections (abscess), and mass effect on the great vessels. Sensitivity and specificity of CT for determination of mediastinal infection were both 100%. In 5 of 10 cases with evidence of mediastinal extension, the clinical situation required surgical drainage. In the other five cases the patients were managed with antibiotic therapy alone. CONCLUSIONS: Mediastinal involvement of chest wall infections is a common occurrence. Conservative and/or surgical management must be tailored to the patient. Factors to consider in management decisions include the clinical status, CT findings, and patient compliance. Rapid CT evaluation may permit timely surgical intervention, limiting complications in some patients. PMID- 12757315 TI - Radiation issues with multidetector row helical CT. PMID- 12757316 TI - Role of trail pheromone in foraging and processionary behavior of pine processionary caterpillars Thaumetopoea pityocampa. AB - Although caterpillars of Thaumetopoea pityocamnpa may mark their pathways with silk, this study shows that the material is essential to neither the elicitation nor maintenance of trail-following or processionary behavior. Trail following is dependent upon a pheromone the caterpillars deposit by brushing the ventral surfaces of the tips of their abdomens against the substate. Earlier instars are strongly bound to their trail system; in the laboratory, caterpillars followed circular trails continuously for as long as 12 hr before breaking away from them. The trail pheromone is long-lived and soluble in nonpolar solvents, but its volatilization or degradation allows the caterpillars to distinguish new from aged trails. In contrast to trail following, processionary behavior, the head-to tail, single-file movement of the caterpillars is dependent on neither silk nor the trail pheromone. Stimuli associated with setae found on the tip of the abdomen of the precedent caterpillar serve to hold processions together, and such stimuli take priority over those associated with either the trail pheromone or silk. Although the caterpillars discern trail strength and choose stronger over weaker trails, the trail marking system of the processionary caterpillar appears less sophisticated than those of other, previously studied species of social caterpillars, and colonies are relatively inefficient in abandoning exhausted feeding sites in favor of new food finds. In laboratory studies, females were more likely to lead processions than males, and leaders, regardless of gender, expended more energy in locomotion than followers. PMID- 12757317 TI - Role of contact pheromones in mate recognition in Xylotrechus colonus. AB - Adult male and female rustic borers, Xylotrechus colonus F. (Coleoptera: Cerambycidae), aggregate on cut logs and fallen trees that are the hosts of their larvae. Our studies show that male X. colonus actively search for females, and only respond to them after contacting them with their antennae. Stripping cuticular hydrocarbons from females with solvent rendered them unattractive to males, suggesting that males did not recognize females by mechanoreception alone. Reapplying solvent extract to washed females restored their attractiveness to males, confirming the role of cuticular hydrocarbons in mate recognition. Female cuticular hydrocarbon extracts contain n-pentacosane, 9-methylpentacosane, and 3 methylpentacosane, components that were either absent or present in very small amounts on males. We demonstrate that the contact pheromone is a blend of these three cuticular hydrocarbons. PMID- 12757318 TI - Contact chemosensory cues in egg bundles elicit male-male agonistic conflicts in the squid Loligo pealeii. AB - Male Loligo pealeii engage in frequent agonistic bouts to gain access to female mates while aggregated at communal egg beds. Male squids are attracted to eggs in the field and in the laboratory. It was recently demonstrated that visual detection followed by physical contact with egg capsules elicited male-male aggression. We tested specific physical and chemical features of the egg capsules that may cause this strong behavioral reaction. Male squids were presented with either natural or artificial egg stimuli and scored for four selected behaviors (egg touch, egg blowing, forward-lunge grab, and fin-beating), the last two of which are highly aggressive behaviors. First, squids were presented with natural eggs versus eggs sealed in agarose-coated tubes (ESACT), which eliminated both tactile and chemical stimuli. Second, males were presented with natural eggs versus eggs sealed in agarose coated tubes containing C18 Sep-Pak-purified extracts (TCPE) from squid egg capsules, which provided chemical cues from natural eggs without the physical stimulus of the egg capsules. Third, natural eggs versus heat-denatured eggs were tested to determine whether the active factor in natural eggs is heat-labile. Squids responded aggressively when contacting natural eggs and TCPE, whereas squids did not respond after touching ESACT or denatured eggs. These results suggest that aggressive behavior is elicited by a heat-labile factor that is embedded within squid egg capsules. This chemosensory cue appears to be a contact pheromone that stimulates the agonistic interactions that characterize the mating behavior of migratory squids on inshore spawning grounds. PMID- 12757319 TI - Male bugs modulate pheromone emission in response to vibratory signals from conspecifics. AB - Male Nezara viridula emit a volatile sex pheromone that acts as a long-range attractant to females. Both sexes communicate through vibrations once they are on the same plant. Males respond to the female calling song by emitting a male courtship song, and they orient to the female calling song on a plant. Simultaneity between vibratory and chemical communication during the last steps of mate finding suggests that pheromone emission might be modulated by signals from conspecifics. Male volatile emissions were collected with fibers for solid phase microextraction, while male bugs were stimulated with natural and artificial signals. Percentages of males releasing pheromone and collected amounts of pheromone increased when males were stimulated with female calling song. Pheromone emission was stable in males stimulated with male rivalry songs, and it decreased in males stimulated with a 100-Hz artificial signal. The ability of male bugs to modulate their pheromone emission may reduce metabolic costs, reduce parasitism, and offer a better synchronization of sexual activity. PMID- 12757320 TI - Characteristic odor of Osmoderma eremita identified as a male-released pheromone. AB - Osmoderma eremita (Scopoli) is an endangered scarab beetle living in hollow trees. It has mainly been known for its characteristic odor, typically described as a fruity, peachlike or plumlike aroma. The odor emanating from a single beetle can sometimes be perceived from a distance of several meters. In this paper, we show that the characteristic odor from O. eremita is caused by the compound (R) (+)-gamma-decalactone, released in large quantities mainly or exclusively by male beetles. Antennae from male and female beetles responded in a similar way to (R) (+)-gamma-decalactone in electroantennographic recordings. Field trapping experiments showed that (R)-(+)-gamma-decalactone is a pheromone attracting female beetles. Lactones similar to (R)-(+)-gamma-decalactone are frequently used as female-released sex pheromones by phytophagous scarabs. This is, however, the first evidence of a lactone used as a male-produced pheromone in scarab beetles. We propose that the strong signal from males is a sexually selected trait used to compete for females and matings. The signal could work within trees but also act as a guide to tree hollows, which are an essential resource for O. eremita. Males may, thus, attract females dispersing from their natal tree by advertising a suitable habitat. This signal could also be exploited by other males searching for tree hollows or for females, which would explain the catch of several males in our traps. PMID- 12757321 TI - (Z,Z)-6,9-heneicosadien-11-one, labile sex pheromone of the whitemarked tussock moth, Orgyia leucostigma. AB - The whitemarked tussock moth (WMTM), Orgyia leucostigma (J. E. Smith), is a major pest of coniferous and deciduous trees in eastern Canada. Chemical identification of its sex pheromone depended primarily on GC-EAD and HPLC analysis, with confirmation of behavioral activity by wind tunnel and field tests. We identified (Z,Z)-6,9-heneicosadien-11-one (Z,Z-6,9-ket) at 4-5 ng/female as the only essential sex pheromone component. Also detected in female extracts were (Z)-6 heneicosen-11-one (Z6-ket) at 2.5 ng/female, (Z,E)-6,8-heneicosadien-11-one (Z,E 6.8-ket) at about 0.5 ng/female, and a trace amount of (Z,E)-6,9-heneicosadien-11 one. Traps containing as little as 1 microg of Z,Z-6,9-ket attracted males at low population levels, indicating it is a potent sex attractant. Traps baited with Z6 ket attracted few males, and in windtunnel bioassays it was at least 100-fold less attractive to males than Z,Z-6,9-ket. No improvement in trap catch occurred with the addition of Z6-ket in various binary mixtures with Z,Z-6,9-ket, including the female ratio, and a ternary mixture of Z,Z-6.9-ket, Z6-ket, and Z,E 6,8-ket in the 9:5:1 ratio detected in females was no better than Z,Z-6,9-ket alone. We attribute the presence of Z,E-6,8-ket and Z,E-6,9-ket in female extracts to the spontaneous and rapid stereospecific isomerization of Z,Z-6,9-ket at room temperature. Male flight began at sunset but peaked during the second half of the night. PMID- 12757322 TI - Evidence for a short-range sex pheromone in female Maladera matrida beetle. AB - Laboratory studies with live and frozen Maladera matrida female and male beetles showed that males were attracted to chemical substances emanating from the females at dusk. Beetles exhibited sexual activity (including mating) at dusk towards frozen females but not towards frozen males. Frozen females that had been extracted with methanol together with either hexane or dichloromethane or with a mixture of all three solvents did not elicit male sexual activity. Activity was fully restored when a concentrate of the extract was applied to the previously extracted frozen females. Males also responded with vigorous sexual activity to frozen males to which female extract had been applied. Deterrent chemicals appear to be absent from the male body. Males exposed to females that had been frozen during the morning displayed weak sexual activity, indicating that females lack active semiochemicals. Differences between dusk and morning extracts were found with respect to more than 20 compounds, some of which were present in much higher concentrations at dusk than in the morning, while others were not detected in the morning extract. The active component(s) of the short-range sex pheromone of female M. matrida is (are) presumably to be found among these compounds. PMID- 12757323 TI - Cuticular lipids of the booklouse, Liposcelis bostrychophila: hydrocarbons, aldehydes, fatty acids, and fatty acid amides. AB - The booklouse, Liposcelis bostrychophila, is an increasingly common pest of stored food products worldwide. We report here the cuticular lipid composition of this pest (the first report of the hydrocarbons of any member of the Order Psocoptera and the first report of fatty acid amides as cuticular components for any insect). No unsaturated hydrocarbons were present. A homologous series of n alkanes (C21-C34), monomethyl alkanes (3-, 4-, 5-, 7-, 9-, 11-, 12-, 13- and 15 methyl-) with a carbon chain range of C28-C42, and dimethyl alkanes (3, 7-; 9, 13 ; 11, 15-; 13, 17-; 9, 21-; 11, 19-; and 13, 21-) with a carbon number range of C31-C41 were identified. The relative abundances of these hydrocarbons were low, comprising approximately 0.0125% of total biomass. The amides were a homologous series (C16-C22 in chain length), with the major amide being stearoyl amide. In addition to the amides, free fatty acids (C16:1, C16:0, C18:2, C18:1, and C18:0 in chain length) and three straight chain aldehydes (C15, C16, and C17:1 in chain length) also occurred as cuticular components. These findings are discussed in terms of the chemical and physiological ecology of this species. PMID- 12757324 TI - Chemical composition of precloacal secretions of two Liolaemus fabiani populations: are they different? AB - The chemical composition of secretions of precloacal pores from two populations of the lizard Liolaemus fabiani (Puilar and Punta-Brava) were compared. This is an endemic species from the Atacama Salt Flat (26 degrees 46'S 68 degrees 14'W; 2400 m) in northern Chile, restricted to the internal lakes of the salt flat. Interpopulational differences in the chemical composition of the secretions were expected considering that populations have genetic differences and are subject to different thermal conditions. By using GC-MS, a total of 44 compounds were found belonging to three categories: n-alkanes, long-chain carboxylic acids, and steroids. Six compounds were found in all the individuals studied: tetradecanoic, pentadecanoic, hexadecanoic, hexadecenoic, octadecanoic, and octadecenoic acids. The secretions of both populations had similar types and proportions of the different compounds, except for cholesterol and hexanoic acid, which were more abundant in the Puilar population. These differences can be ascribed to the different environmental conditions to which populations are subjected. The adaptive meaning of these differences is discussed. PMID- 12757325 TI - Interspecific variation in terpenoid composition of defensive secretions of European Reticulitermes termites. AB - Sixteen terpene compounds were isolated from the soldier defensive secretions of seven European termite taxa of the genus Reticulitermes (Isoptera, Rhinotermitidae). We describe species-specific mixtures of monoterpenes (alpha pinene, beta-pinene, limonene), sesquiterpenes (germacrene C, germacrene A, germacrene B, beta-selinene, delta-selinene, gamma-selinene, (E)-beta-farnesene, gamma-cadinene, nerolidol), diterpenes (geranyl linalool, geranyl geraniol, geranyl geranial), and one sesterterpene (geranyl farnesol). Compounds were purified by HPLC and their structures determined by means of MS spectrometry, or 1D and 2D NMR spectroscopy. Comparison of two different analytical approaches, CC MS and HPLC with subsequent NMR spectroscopy, revealed Cope rearrangement of germacrene A, germacrene B, and germacrene C to the respective beta-elemene, gamma-elemene, and delta-elemene under GC conditions, thus demonstrating the limits for this analytical approach. The species-specific compound composition provides insight into taxonomy and species origin of European Reticulitermes. The biological significance of the species-specific composition of Reticulitermes defensive secretions is briefly discussed. PMID- 12757326 TI - Combined roles of contact stimulant and deterrents in assessment of host-plant quality by ovipositing zebra swallowtail butterflies. AB - Zebra swallowtail (Eurytides marcellus) butterflies are stimulated to oviposit by a single compound, 3-caffeoyl-muco-quinic acid (1). Analysis of the aqueous extracts of the leaves of the host. Asimina triloba, showed that they contained stimulant 1, its isomer (2), and the flavonoids rutin (3) and nicotiflorine (4) as major components. We compared the concentrations of compounds 1-4 in terminal leaves (TL) and expanded leaves (EL) of the host plants at four different times throughout the growing season. In spring, the concentration of 1 was highest in TLs, and flavonoids were not detectable or present at low levels. As the season progressed, however, the concentrations of flavonoids increased, reached maxima by late summer, and then decreased as the plants started senescing. There were also significant differences in the concentrations of these compounds between TLs and ELs. In a choice assay with model leaves, we tested equivalent amounts of post-dichloromethane aqueous extracts made in spring (May) and in fall (September). September extracts received significantly fewer approaches and eggs. In greenhouse experiments with potted A. triloba plants, the butterflies chose some leaves to lay eggs, while others were rejected or ignored. Analyses showed that the concentrations of compound 1 were not significantly different in the three kinds of leaves. The flavonoids (3 and 4), however, were significantly higher in the leaves that were ignored. Multiple-choice tests using model plants suggested that concentrations of both flavonoids and stimulant were important in assessing host suitability. There was a gradual decrease in approaches as the concentration of 1 decreased. Higher amounts of flavonoids deterred egg laying even in the presence of high concentrations of stimulant 1. At lower concentrations of 1, the addition of low doses of flavonoids deterred egg laying. Thus, the results suggest that the butterflies use both qualitative and quantitative information about these compounds to assess host quality. This behavior may have evolved to take advantage of seasonal variation in the chemistry of their host, A. triloba. PMID- 12757327 TI - Characterization of furanocoumarin metabolites in parsnip webworm, Depressaria pastinacella. AB - Although metabolites of furanocoumarins have been characterized in a wide range of organisms, to date they have been identified in only a single insect species, Papilio polyxenes. Depressaria pastinacella, the parsnip webworm, like P. polyxenes a specialist on Apiaceae, routinely consumes plant tissues higher in furanocoumarin content than does P. polyxenes and is capable of faster cytochrome P-450-mediated detoxification of these compounds. In this study, we characterized metabolites of xanthotoxin, a linear furanocoumarin, and sphondin, an angular furanocoumarin, in midguts and frass of parsnip webworms. Two metabolites were isolated and identified from webworms fed artificial diet containing xanthotoxin. LC-ESI-MS analysis resulted in the determination of a MW of 266 for the compound in the frass and one of the compounds in the midgut; 1H NMR confirmed its structure as 6-(7-hydroxy-8-methoxycoumaryl)-hydroxyacetic acid (HCHA). The second compound from the midgut had a MW of 252 and was identified by 1H NMR and 13C NMR analysis as 6-(7-hydroxy-8-methoxycoumaryl)-hydroxyethanol) (HMCH). Whereas HCHA has been found in frass of Papilio polyxenes fed xanthotoxin, HMCH has not been reported previously in insects. Although the first step of metabolism of xanthotoxin in webworms as well as P. polyxenes is likely the formation of an epoxide on the furan ring, angular furanocoumarin metabolism in webworms appears to differ. The principal metabolite of sphondin was identified as demethylated sphondin (6-hydroxy-2H-furo[2,3-h]-1-benzopyran-2-one) by LC-ESI MS and confirmed by 1H NMR and 13C NMR analyses. That webworms produce metabolites of xanthotoxin in common not only with other Lepidoptera (e.g., HCHA) but with other vertebrates (e.g., HMCH) suggests a remarkable conservatism in the metabolic capabilities of cytochrome P-450s and raises the possibility that insects may share other detoxification reactions with vertebrates with respect to toxins in foodplants. PMID- 12757328 TI - Antioxidants in grasshoppers: higher levels defend the midgut tissues of a polyphagous species than a graminivorous species. AB - Polyphagous grasshoppers consume plants that contain markedly greater amounts of potentially prooxidant allelochemicals than the grasses eaten by graminivorous grasshoppers. Therefore, levels of antioxidant defenses maintained by these herbivores might be expected to differ in accordance with host plant ranges. Antioxidant levels were compared in midgut tissues and gut fluids of a polyphagous grasshopper. Melanoplus sanguinipes, and a graminivorous grasshopper, Aulocara ellioti. Glutathione concentrations in midgut tissues of M. sanguinipes (10.6 mM) are among the highest measured in animal tissues and are twice as high as those in A. ellioti. Alpha-tocopherol levels are 126% higher in midgut tissues of M. sanguinipes than in those of A. ellioti, and remain at high levels when M. sanguinipes is reared on plants containing a wide range of alpha-tocopherol concentrations. Ascorbate levels in M. sanguinipes midgut tissues are 27% higher than in those of A. ellioti, but vary depending on the host plant on which they are reared. Midgut fluids of both species contain elevated levels of glutathione, as well as large (millimolar) amounts of undetermined antioxidants that are produced in the insects. The consumption of tannic acid decreases ascorbate concentrations in midgut tisssues and gut fluids of A. ellioti but has no effect on ascorbate levels in M. sanguinipes. The results of this study provide the first measurements of antioxidants in grasshoppers and suggest that the maintenance of high levels of antioxidants in the midgut tissues of polyphagous grasshoppers might effectively protect them from oxidative stress. PMID- 12757329 TI - Linking chemical reactivity and protein precipitation to structural characteristics of foliar tannins. AB - Tannins influence ecosystem function by affecting decomposition rates, nutrient cycling, and herbivory. To determine the role of tannins in ecological processes, it is important to quantify their abundance and understand how structural properties affect reactivity. In this study, purified tannins from the foliage of nine species growing in the pygmy forest of the northern California coast were examined for chemical reactivity, protein precipitation capacity (PPC), and structural characteristics (13C NMR). Reactivity of purified tannins varied among species 1.5-fold for the Folin total phenol assay, and 7-fold and 3-fold, respectively, for the acid butanol and vanillin condensed tannin assays. There was about a 5-fold difference in PPC. Variation in chemical reactivity and PPC can be largely explained by differences in structural characteristics of the tannins determined by 13C NMR. In particular, the condensed versus hydrolyzable tannin content, as well as the hydroxylation pattern of the B-ring and stereochemistry at the C-2-C-3 position appear to influence reactivity. Due to the large differences in chemical reactivity among species, it is necessary to use a well-characterized purified tannin from the species of interest to convert assay values to concentrations. Our results suggest that structural characteristics of tannins play an important role in regulating their reactivity in ecological processes. PMID- 12757330 TI - Effects of molasses grass, Melinis minutiflora volatiles on the foraging behavior of the cereal stemborer parasitoid, Cotesia sesamiae. AB - Olfactory responses of the cereal stemborer parasitoid Cotesia sesamiae to volatiles emitted by gramineous host and nonhost plants of the stemborers were studied in a Y-tube olfactometer. The host plants were maize (Zea mays) and sorghum (Sorghum bicolor), while the nonhost plant was molasses grass (Melinis minutiflora). In single-choice tests, females of C. sesamiae chose volatiles from infested and uninfested host plants and molasses grass over volatiles from the control (soil). In dual-choice tests, the wasp preferred volatiles from infested host plants to those from uninfested host plants. There was no discrimination between molasses grass volatiles and those of uninfested maize, uninfested sorghum, or infested maize. The wasp preferred sorghum volatiles over maize. Combining uninfested maize or sorghum with molasses grass did not make volatiles from the combination more attractive as compared to only uninfested host plants. Infested maize alone was as attractive as when combined with molasses grass. Infested sorghum was preferred over its combination with molasses grass. Local growth conditions of the molasses grasses influenced attractiveness to the parasitoids. Volatiles from Thika molasses grass were attractive, while those from Mbita molasses grass were not. Growing the Thika molasses grass in Mbita rendered it unattractive and vice versa with the Mbita molasses grass. This is a case of the same genotype expressing different phenotypes due to environmental factors. PMID- 12757331 TI - Effect of urea fertilizer application on soluble protein and free amino acid content of cotton petioles in relation to silverleaf whitefly (Bemisia argentifolii) populations. AB - The impact of urea nitrogen fertilization on silverleaf whitefly, Bemisia argentifolii Bellows & Perring, population dynamics was examined in field-grown cotton (Gossypium hirsutum L.). Five urea nitrogen treatments were tested, consisting of soil applications of 0, 112, 168, and 224 kg nitrogen per hectare, and acombined soil-foliar application of 112:17 kg nitrogen per hectare. A positive response was observed between N application rates and the measured levels of nitrate N in petioles from mature cotton leaves. Similarly, a positive response was observed between N application rates and the numbers of adult and immature whiteflies appearing during population peaks. To determine whether these positive responses were related, we measured the levels of dietary N compounds (proteins and free amino acids) that would be available for insect nutrition in cotton petioles at the different N application rates. Sampling dates and N application treatments affected levels of soluble proteins in cotton petioles, and interactions between sampling dates and treatments were significant. Across all sampling dates, the relationship between N application rates and levels of soluble proteins was linear. Sampling dates also affected levels of total and individual free amino acids. Fertilizer treatments only affected levels of total amino acids, aspartate, asparagine, and arginine plus threonine. Levels of aspartate or asparagine and the N application rates were linearly correlated. No significant correlations were observed between levels of dietary N compounds in cotton petioles and numbers of whiteflies, either adults or immatures, on the cotton plants. PMID- 12757332 TI - Constraints to herbivore-induced systemic responses: bidirectional signaling along orthostichies in Nicotiana attenuata. AB - We investigated the impact of leaf vascular connections on systemically transmitted herbivore-induced gene expression in Nicotiana attenuata. Although systemic signaling is clearly associated with the plant vascular system, few studies consider vascular architecture when measuring systemically induced defenses. N. attenuata is a plant with dispersed phyllotaxis approximating 3/8 in the rosette stage of growth. We mimicked Manduca sexta herbivory by introducing larval regurgitant to wounds produced with a standardized continuous mechanical wounding and investigated mRNA accumulation of genes. Herbivory in N. attenuata induces the expression of genes coding for a proteinase inhibitor protein (PI), threonine deaminase (TD, EC 4.3.1.19), a luminal-binding protein (BiP), and an alpha-dioxygenase (alpha-DOX). We measured the systemic response of sink leaves when orthostichous (growing at an angular distance of 0 degrees) source leaves were treated, and vice versa, and compared it to the systemic response of leaves growing at the maximum angular distance of 180 degrees. Vascular architecture clearly controlled the intensity of systemic mRNA accumulation within the 4-hr time frame of the experiment. In addition, we found signal translocation to be bidirectional, travelling from source to sink as well as from sink to source leaves, which argues against a phloem-based assimilate-linked signal identity. PMID- 12757333 TI - Distribution of the carcinogenic terpene ptaquiloside in bracken fronds, rhizomes (Pteridium aquilinum), and litter in Denmark. AB - The distribution of ptaquiloside (PTA) was studied in four Danish bracken populations in order to evaluate the transfer of PTA from ferns to soil. Populations showed statistically significant differences in PTA contents of fronds and rhizomes despite large in-site variations. The highest concentrations were encountered in fronds with concentrations ranging between 213 and 2145 microg/g, while rhizomes had concentrations between 11 and 902 microg/g. PTA was present in soil materials in amounts of 0.22-8.49 microg/g but apparently with no correlation with PTA contents of fronds or rhizomes. Laboratory tests showed that water could leach PTA from bracken fronds, which is in support of the high soil contents observed at sites exposed to heavy showers just before sampling. The observed soil contents correspond to estimated soil solution concentrations of 200-8500 microg/liter, demonstrating a substantial risk of PTA contamination of surface water and groundwater. PMID- 12757334 TI - Left ventricular aneurysmectomy: endoventricular circular patch plasty or septoexclusion. AB - BACKGROUND: Septoexclusion is a technique described by Guilmet in the mid 1980s. Its indications and midterm results are evaluated and compared to those obtained with the Dor operation. METHODS: From January 1998 to April 2001, 79 patients had an exclusion of scars following myocardial infarction in left anterior descending artery (LAD) territory. Fifty of them (63.3%) had the Dor operation (Group D) and 29 (36.7%) the Guilmet operation (Group G). Dor technique was used when the involvement of the septum and the free wall was roughly similar. Guilmet technique was indicated when the septum was involved at a greater extent than the free wall. Ejection fraction (EF) was lower and end-diastolic volumes were higher in Group G. Incidence of functional mitral regurgitation was similar in both groups. RESULTS: Thirty-day mortality was 7.6% (8.0% in Group D versus 6.9% in Group G, p = ns). After a mean of 21.0 +/- 8.5 months, five patients (6.9%) died, two in Group D and three in Group G. Causes of death were cardiac related in four and not cardiac related in one. Mean follow-up of the 68 survivors was 24.3 +/- 12.0 months (range: 4-38 months). Fifty patients (73.5% of the survivors) improved (28 in Group D and 22 in Group G, p = 0.026), whereas in 18, New York Heart Association (NYHA) class remained unchanged or worsened. Both groups showed an increase of EF and a volumetric reduction, whereas stroke volume remained unchanged. Fewer patients had mitral regurgitation than in the preoperative period (41.3% versus 65.8%, p = 0.013) and at a lesser extent (1.7 +/- 0.7 versus 0.7 +/- 0.6, p < 0.001). CONCLUSIONS: Our results show that both Dor and Guilmet techniques are effective in the surgical treatment of left ventricular dyskinetic or akinetic areas related to LAD territory. Each technique has its own indications and has to be addressed to patients with different extension of septal scars. PMID- 12757335 TI - Predictors of early outcome after coronary artery surgery in patients with severe left ventricular dysfunction. AB - BACKGROUND: The surgical survival in patients with severe myocardial dysfunction is critically dependent on the selection of patients. The present study was undertaken to identify the prognostic factors in such patients. METHODS: We analyzed the data of 176 consecutive patients (161 men, 15 women), aged 29 to 88 years (mean 58.43), with a left ventricular ejection fraction (LVEF) < 30% who underwent isolated coronary artery bypass grafting. The LVEF ranged from 15% to 30% (mean 27.18%). Preoperatively, 33% had angina, 19.9% had recent myocardial infarction, and 21.6% had congestive heart failure. The mean number of grafts was 2.5/patient. The intra-aortic balloon was used prophylactically in 20.5% of patients and therapeutically in 4.0% of patients. RESULTS: The hospital mortality was 2.3%. The complications occurred as follows: perioperative myocardial infarction in two (1.1%), intractable ventricular arrhythmias in two (1.1%), prolonged ventilation in four (2.3%) and peritoneal dialysis in 1 (0.6%). The mean ICU and hospital stay were 2.46 +/- 0.76 and 7.57 +/- 2.24 days, respectively. The predictors of survival on univariate analysis were New York Heart Association (NYHA) class (x2 = 14.458, p < 0.001), recent myocardial infarction (x2 = 5.852, p = 0.016), congestive heart failure (CHF) (x2 = 5.526, p = 0.019), and left ventricular end-systolic volume index (LVESVI) (x2 = 25.833, p < 0.001). However, on multivariate analysis, left ventricular end-systolic volume index was the only independent left ventricular function measurement predictive of survival (x2 = 10.228, p = 0.001). CONCLUSION: Left ventricular end-systolic volume index is the most important predictor of survival after coronary artery bypass surgery in patients with severe myocardial dysfunction. PMID- 12757337 TI - Surgical treatment of post infarction left ventricular aneurysms: our experience with double breasting and Dor's repair. AB - BACKGROUND: This is a retrospective study of left ventricle (LV) aneurysm repair done at the Escorts Heart Institute and Research Centre, New Delhi, since October 1988. Two methods of LV aneurysm repair are practiced: double breasting (DB) and Dor's repair. The method varies with location, size, and extent of aneurysm and quality of the fibrotic wall of the aneurysm. METHODS: Between October 1988 and May 2001, 129 patients underwent LV aneurysm repair using one of the two techniques; 78 patients had Dor's repair while 51 patients had DB repair. RESULTS: Overall mortality was 2.3% (three patients). One patient died in the DB group, and two patients died in the Dor's repair group. Mean preoperative ejection fraction (EF) after surgery in DB was 31% while in Dor's repair it was 29.2%. EF showed improvement after surgery to 48.5% in DB and 46.6% in Dor's repair. Decrease in end-diastolic volume (EDV) in DB was from 146 to 91.4 cm3/m2, and in Dor's repair it was from 156 cm3/m2 to 88.6 cm3/m2. Decrease in end systolic volume was from 101 cm3/m2 to 60.2 cm3/m2 in DB and from 109 cm3/m2 to 64.5 cm3/m2 in Dor's group. All of these values showed statistically significant improvement. At six months postoperatively, 83 patients (74.1%) out of 112 patients who were preoperatively in New York Heart Association (NYHA) Functional Classes III and IV improved to class II while 7 patients (6.3%) improved to class I. CONCLUSION: In our experience Dor's repair is indicated for anteroseptal and apical isolated posterior aneurysm to restore LV volume and geometry while DB is indicated for apical, anterolateral, and lateral aneurysms where septal involvement is less. These two techniques have definite indications and advantages with good results. PMID- 12757336 TI - Left ventricular assist devices as bridge to heart transplantation: The Niguarda Experience. AB - BACKGROUND: Congestive heart failure is the leading cause of death in Western countries. Heart transplantation currently is the only accepted therapy for patients with end-stage heart failure, but the supply of donor hearts is inadequate, and different mechanical circulatory support systems have been investigated as bridges to heart transplant. METHODS: Since April 1992, 53 patients (47 men, 6 women, aged 12 to 61 years) received left ventricle mechanical circulatory support as bridge to heart transplant. The two principal devices used were: the Novacor LVAS in 31 patients and the DeBakey VAD in 11 patients. RESULTS: All patients survived the operation. Mean duration of LVAD support was 2.8 +/- 5.6 months. Thirty-seven patients (71.1%) underwent heart transplantation. Twelve major bleeding episodes occurred in nine patients (16.9%). Globally, major and minor neurologic events occurred in 13 patients (24.5%). Ten patients (19.9%) assisted with the Novacor Wearable LVAS device were discharged at home while waiting for heart transplant (HTx). The mean follow-up of the 34 discharged transplanted patients was 45.3 +/- 37 months. Actuarial survival of transplanted patients while on LVAD was 91.0 +/- 4.9% and 83.4 +/- 8.5% at 1 and 5 years, respectively. No differences in post-transplant long-term survival and rejection and allograft vasculopathy occurred between patients transplanted with or without LVAD implanted. CONCLUSIONS: LVAD therapy proved to be effective in bridging patients with end-stage heart failure to HTx. While on LVAD support, patients assisted with implantable wearable devices could be discharged at home, ameliorating their quality of life. The excellent survival rate after HTx is concomitant with a low incidence of rejection and cardiac allograft vasculopathy. PMID- 12757338 TI - Left ventriculoplasty for nonischemic dilated cardiomyopathy. AB - From December 1996 to May 2001, we have performed 82 cases of left ventriculoplasty (LVP) for nonischemic dilated cardiomyopathy (DCM). Surgical procedure was partial left ventriculectomy in 70 patients and septal anterior ventricular exclusion in 12 patients with evaluation by utilizing intraoperative echocardiography. There were 70 men and 12 women with a mean age of 49, varying from 14 to 76. All patients had medically refractory heart failure with New York Heart Association (NYHA) Functional Class III in 33 patients and NYHA IV in 49; 34 patients were supported by inotropic infusion prior to the operation. Intra aortic balloon pump (IABP) and left ventricle assist device (LVAD) were used in 12 and 2 patients at perioperative period, respectively. Hospital mortality was 8.2% in elective operation (5/61), 57.1% in emergency operation (12/21), and 20.7% overall. One- and four-year survival rates were 75.5% and 69.3% in elective cases, 37.9% and 0 in emergency cases, and 64.7% and 3.6% overall, respectively. Left ventricular (LV) ejection fraction increased from 22.3% to 29.0% at the time of surgery and has maintained around 33% up to two years. LV diastolic dimension and LV end diastolic pressure decreased from 83.8 mm to 65.0 mm; from 31.7 mmHg to 22.0 mmHg have maintained around 70 mm and 22.1 mmHg up to two years, respectively. Over one year most of the survivors were medically controlled within NYHA Class I-II. In conclusion, careful choice of surgical procedure by utilizing intraoperative echocardiography enables left ventriculoplasty to effectively treat severe heart failure with nonischemic cardiomyopathy. PMID- 12757339 TI - Early in vivo hemodynamic results after aortic valve replacement with the St Jude Medical Regent mechanical heart valve in patients with pure aortic stenosis. AB - BACKGROUND: The St Jude Medical Regent is a new-generation mechanical heart valve that represents a design evolution of the St Jude Hemodynamic Plus Series (HP). The purpose of this study was to evaluate early "in vivo" hemodynamic performance of the Regent valve in patients with aortic stenosis. METHODS: Between March 2000 and December 2001, 32 patients (mean age 59.9 +/- 5.9,56.3% male) with pure aortic stenosis received a Regent mechanical prosthesis in the aortic position. Hemodynamic performance was assessed by Doppler echocardiography at discharge, two months, six months, and one year by calculating peak transprosthetic velocity (Vmax), mean (MG) and peak (PG) transprosthetic gradients, effective orifice area index (EOAI), left ventricular mass index (LVMI), and degree of aortic regurgitation. RESULTS: A significant reduction in mean and peak transaortic gradients (p < 0.001) and a significant increase in EOAI (p < 0.001) over time followed valve replacement, and a bivariate analysis of variance (ANOVA) failed to demonstrate statistical differences by valve size over time (p = ns). A significant reduction in left ventricular hypertrophy occurred over time (p < 0.001) in all valve sizes (p = ns between groups): baseline LVMI was 221 +/- 57 g/cm2; it decreased by 30 g/cm2 (p < 0.001) at discharge. LVMI decreased from 191 +/- 54 g/cm2 to 161 +/- 41 g/cm2 (p < 0.001) from discharge to two months. Further reductions were not significant. At the six-month follow-up no patient in our cohort exhibited moderate or severe aortic regurgitation at Doppler echocardiography. CONCLUSIONS: Early results with the St Jude Medical Regent valve have been satisfactory. Further assessments are necessary to confirm these results. PMID- 12757340 TI - Prospective comparison of minimally invasive and standard techniques for aortic valve replacement: initial experience in the first hundred patients. AB - BACKGROUND: Aortic valve replacement (AVR) can be performed through a partial upper sternotomy. In this study we compared the early postoperative outcome in two groups of patients who underwent AVR with a minimally invasive procedure (n = 30) or with a conventional approach (n = 70). The predicted operative mortality (Parsonnet Index) was slightly higher in the conventional group (17.69 +/- 0.85 versus 12.7 +/- 1.02), reflecting the greater mean age of the patients (70.96 +/- 1.17 versus 64.20 +/- 2.57). RESULTS: The distribution of the different etiologies of aortic valve pathology did not differ between groups. There was no postoperative death in the mini-invasive group. Cardiopulmonary bypass time was longer in the mini-invasive group, but the other operative parameters did not differ between groups. Postoperative morbidity regarding the need for blood transfusion, the duration of assisted ventilation, length of stay in the intensive care unit, and abnormalities of cardiac rhythm and conduction was slightly but not significantly reduced in the mini-invasive group. CONCLUSIONS: Our data demonstrate that a partial upper sternotomy is a safe and effective technique for AVR. Postoperative morbidity is not significantly reduced in patients undergoing AVR by this approach. Further studies in a larger patient population are necessary to assess whether postoperative morbidity is significantly reduced. PMID- 12757341 TI - Heparin-coated circuits (Duraflo II) with reduced versus full anticoagulation during coronary artery bypass surgery. AB - BACKGROUND: Introduction of completely heparin-coated cardiopulmonary bypass (CPB) circuits combined with reduced systemic anticoagulation has been shown to reduce postoperative bleeding and requirements for allogeneic transfusions after cardiac surgery. However, some uncertainty exists whether this effect is due to the reduced amount of heparin or to the heparinized surface itself. Therefore, a retrospective study was undertaken, comparing two different anticoagulation protocols applied to coronary artery bypass patients treated with identical heparin-coated CPB equipment. METHOD: Over a 12 month period all coronary artery bypass patients operated with extracorporeal circulation were subjected to a Duraflo II heparin-coated circuit (Baxter Healthcare Corp, Bentley Laboratories Division, Irvine, Calif) and full heparin dose (activated clotting time [ACT] > 480 seconds; Group F, n = 651). Over the next 24 months, all coronary patients who were treated with an identical circuit combined with reduced systemic heparinization (ACT > 250 seconds) were included in Group R (n = 675). Except for the different anticoagulation protocols, all treatment regimens before, during, and after the operation remained unchanged throughout the study period. RESULTS: There were no statistically significant differences in any major demographic or operative parameters. In Group R, the postoperative bleeding was mean 665 +/- 257 ml versus 757 +/- 367 ml in Group F (p < 0.0001), and the perioperative decrease in hemoglobin concentration was significantly lower in Group R (22 +/- 1.2 gm/L versus 25 +/- 1.3 gm/L, p < 0.0001). The time for postoperative ventilatory support was shorter in Group R (1.7 +/- 1.3 hours versus 1.9 +/- 1.1 hours in Group F, p = 0.0006), and the incidence of new episodes of atrial fibrillation after the operation was lower (26.4% in Group R versus 32.8% in Group F, p = 0.01). There were no significant differences in the incidences of perioperative myocardial infarction, stroke, transient neurological disturbances, physical rehabilitation, or mortality. No technical or coagulation problems were recorded in either group. CONCLUSION: The use of Duraflo II coated circuits for CPB combined with reduced anticoagulation decrease postoperative bleeding and hemoglobin loss compared with full heparin dose treatment. In addition, the intubation time was shorter and the incidence of postoperative atrial fibrillation was lower in the patients treated with low heparin doses. PMID- 12757342 TI - Limits of arterial myocardial revascularization. AB - A prospective study of myocardial blood perfusion after coronary artery bypass graft (CABG) was conducted in two groups of patients. In group 1, a two-year assessment by exercise thallium myocardial scintigraphy without medical treatment was performed in 122 patients who consecutively underwent CABG with exclusive use of both internal mammary arteries (IMA) and gastroepiploic artery (GEA). In group 2, myocardial function and perfusion were determined by radionuclide investigations performed before and one year after CABG in 100 patients with preoperative LV dysfunction (defined as LV ejection fraction (LVEF) less than 0.40), comparing results of myocardial revascularization performed with either exclusive arterial grafts (arterial group, 54 patients) or one arterial graft (IMA) associated with a sequential vein graft (vein group, 46 patients). In group 1, 21% of patients presented silent residual electric ischemia during exercise stress testing and 26% had reversible scintigraphic ischemic defect despite complete revascularization, 18% of those in the inferior wall bypassed with GEA and 8% in the anterior wall bypassed with the right IMA. In group 2, the significant preoperative ischemia significantly decreased in both the vein group and the arterial group. LV function was significantly improved in the vein group; in contrast there was no modification of LV function in the arterial group. A multivariate analysis showed that the surgical technique used and the preoperative LVEF were independent prognostic factors of the postoperative myocardial outcome, with a positive impact of the vein use on the postoperative myocardial function recovery. It is important to recognize that arterial grafts have some limitations in the ability to supply blood flow for coronary circulation that may induce postoperatively silent residual myocardial ischemia and a lack of LV function recovery. PMID- 12757343 TI - Descending aortic dissection post coarctation repair in a patient with Turner's syndrome. AB - Dissection of the descending aorta post coarctation repair in a patient with Turner's syndrome has rarely been described. A 45-year-old woman with Turner's syndrome had repair of coarctation by resection and interposition graft. Her postoperative course was uneventful. Chest X-ray two months postoperatively showed a hematoma in the proximal descending aorta, and a CT scan confirmed dissection distal to the coarctation repair, which was treated medically. Subsequent CT scanning one year later showed the hematoma resolving with no increase in the diameter of the dissected segment. PMID- 12757344 TI - Carbamazepine-related hyponatremia following cardiopulmonary bypass. AB - A 67-year-old man on long-term carbamazepine therapy underwent elective coronary artery bypass grafting. Following an initially uncomplicated recovery, he developed symptomatic hyponatremia. The symptoms and biochemical abnormality improved after gradual discontinuation of carbamazepine. We discuss the association between carbamazepine and hyponatremia and the causes of hyponatremia after cardiopulmonary bypass. Surgeons should consider stopping carbamazepine before operations with cardiopulmonary bypass. PMID- 12757345 TI - Single aortic clamp versus partial occluding clamp technique for cerebral protection during coronary artery bypass: a randomized prospective trial. AB - Single aortic clamp (SAC) versus partial occluding clamp (POC) technique for the construction of proximal anastomosis has been suggested to provide better cerebral protection during coronary artery bypass grafting (CABG). The aim of this study was to assess this hypothesis in a prospective randomized trial. METHODS: Two hundred sixty-eight consecutive patients underwent CABG at a single institution. All patients were randomized to either SAC (Group S) or POC (Group P) for the construction of the proximal anastomosis. Myocardial protection consisted of multidose antegrade cold blood cardioplegia with topical cooling. The operations were performed using standard cardiopulmonary bypass support and moderate systemic hypothermia (29 to 32 degrees C). The incidences of neurological events, perioperative myocardial infarction (MI), and mortality were prospectively evaluated. RESULTS: The two groups were similar in mean age, gender, urgency of operation, and number of bypasses. Group S patients had a significantly longer cross-clamp (61 +/- 21 minutes [S] vs 44 +/- 13.8 minutes [P], p < 0.05) and bypass times (85 +/- 25 minutes [S] vs 74 +/- 19.7 minutes [P], p < 0.05). There were no differences in the number of perioperative MIs (Group S = 3 [2.3%]; Group P = 2 [1.5%], p = 0.50) or mortality (Group S = 2 [1.5%]; Group P = 3 [2.2%], p = 0.50). Two patients randomized to POC were switched to SAC intraoperatively because of severe calcification of the ascending aorta. In Group P, there were two strokes (1.5%) and two (1.5%) postoperative confusions versus none in Group S (relative risk = 2.0, p < 0.05, respectively). CONCLUSION: The SAC technique improved cerebral protection without any adverse effect on myocardial protection and postoperative outcome in patients undergoing CABG. PMID- 12757346 TI - Two cases using "epi-endocardial patch repair" for postinfarction left ventricular rupture. AB - The treatment points of left ventricular (LV) free wall rupture after acute myocardial infarction (MI), so far, are to prevent a deterioration of LV function after MI and to prevent a recurrence or extension of the dissection of the infarcted/necrotic myocardium to stop bleeding. We report two cases of LV rupture after myocardial infarction that underwent epicardial patch repair using deep epicardial sutures reaching LV subendocardial area ("epi-endocardial patch" repair). The procedure was done under beating condition with cardiopulmonary bypass in the first case and with preoperatively percutaneous cardiopulmonary support system (PCPS) in the second case to prevent a deterioration of LV function. Hemostasis was effective and complete, and extension of the intramuscular dissection was well blocked. The patients recovered LV function soon. The epi-endocardial sutures can be placed safely without inducing new ischemia, and the method might be possible with beating condition. PMID- 12757347 TI - Impending paradoxical embolism after coronary artery bypass grafting successful surgical treatment. AB - We describe a case of impending paradoxical embolism of a 22 cm long thromboembolus, straddling over a patent foramen ovale, detected by transthoracic and transesophageal echo 11 days after a coronary artery bypass operation. The patient underwent successful emergency removal of the clot and closure of the patent foramen ovale. A vena cava filter was placed because of new thrombi detected in deep veins of the legs. PMID- 12757348 TI - Beating heart surgery in a patient with dextrocardia and complete situs inversus. AB - We present a 65-year-old female patient with dextrocardia and situs inversus who underwent successful coronary artery bypass without cardiopulmonary bypass. Vessels revascularized included right internal mammary artery to the left anterior descending artery and a saphenous vein graft to the first obtuse marginal branch. The procedure was performed on a beating heart through a median sternotomy with the use of a compression epicardial stabilizer. The patient was discharged to her home after an uneventful recovery. Only 12 similar cases of myocardial revascularization in patients with dextrocardia have been reported so far, and this is one of the first procedures, in patients with dextrocardia, performed off pump. PMID- 12757349 TI - Surgical approach to ascending aorta in bicuspid aortic valve. AB - OBJECTIVE: Former studies have pointed out that hemodynamic stress imposed by associated valvular disease is the primary factor in the development of ascending aorta dilatation. At present, intrinsic wall pathology is blamed for dilatation and aneurysm formation in bicuspid aortic valve (BAV). MATERIALS AND METHODS: Aortic valve replacement (AVR) was performed on 78 adult patients with BAV. Patients were divided into two groups. Group I (n = 27) underwent only AVR. Group II (n = 51) underwent AVR and additional ascending aorta procedures such as Shawl Lapel aortoplasty (n = 12) and tailoring aortoplasty (n = 9). Dacron wrapping was performed after both techniques were done. Ascending aorta replacement was done on 11 patients by using composite graft. Supracoronary graft replacement was performed in 3 patients after AVR. RESULTS: Ascending aorta diameter increment was 1.25 mm/year in normotensive and 2.80 mm/ year in hypertensive patients. Ascending aorta aneurysm (diameter > 55 mm) developed in eight patients in the postoperative period in group I. Ascending aorta dilatation did not develop in group II patients. Mean survival time +/- standard error (SE) was 128 +/- 11 and 99 +/- 4 months and survival possibility was 77.78% and 92.16%. Freedom from reoperation was 65.4% and 95.9% in 8 years in group I and group II, respectively. CONCLUSION: Aortic wrapping with or without aortoplasty has a beneficial effect not only in dilated ascending aorta but also in all nondilated BAV patients with normal-sized aortic diameter. Ascending aorta wrapping in BAV patients preserves the endothelial lining and prevents further dilatation, aneurysm formation, and dissection. PMID- 12757351 TI - General education teachers' relationships with included students with autism. AB - In this study, we examine the relationship between general education teachers and second- and third-grade included students with autism. We also examine the effect of childrens' behavior problems on these relationships, as well as inclusion within the social environment of the classroom. Included students with autism form multidimensional relationships with their general education teachers. These relationships are associated with student's display of behavior problems and level of inclusion in the class. Specifically, when teachers perceived their relationships with included students with autism to be more positive, children's levels of behavioral problems were lower and they were more socially included in the class. PMID- 12757350 TI - Responses and sustained interactions in children with mental retardation and autism. AB - Sustained interactions and responses to social bids made by children with autism and verbal-age-matched children with mental retardation were recorded in two naturalistic settings. Children with autism produced fewer positive responses and more "no responses" than children with mental retardation; both groups were more likely to make positive responses to adults and not to respond to other children. Furthermore, although the frequency of conversations was not different for the two groups, children with autism were significantly less likely to engage in sustained play compared to children with mental retardation. Results suggest that children with autism are able to master the more rote and need-oriented social skills, such as simple conversation, but may not develop other forms of social interactions, like play. PMID- 12757352 TI - Effects of a model treatment approach on adults with autism. AB - The study evaluated the effectiveness of a residential program, based on the TEACCH model, in improving the quality of the treatment program and the adaptation of individuals with autism with severe disabilities. The results indicated that participants in the Carolina Living and Learning Center experienced an increase in structure and individualized programming in the areas of communication, independence, socialization, developmental planning, and positive behavior management compared to participants in control settings. The experimental program was viewed as a more desirable place to live than the other settings, and the families were significantly more satisfied. Based on exploratory analyses, the use of the TEACCH methods over time were related to a decrease in behavior difficulties. There was no difference in the acquisition of skills. PMID- 12757353 TI - Behavioral adjustment of siblings of children with autism engaged in applied behavior analysis early intervention programs: the moderating role of social support. AB - There have been few studies of the impact of intensive home-based early applied behavior analysis (ABA) intervention for children with autism on family functioning. In the present study, behavioral adjustment was explored in 78 siblings of children with autism on ABA programs. First, mothers' ratings of sibling adjustment were compared to a normative sample. There were no reported increases in behavioral adjustment problems in the present sample. Second, regression analyses revealed that social support functioned as a moderator of the impact of autism severity on sibling adjustment rather than a mediator or compensatory variable. In particular, siblings in families with a less severely autistic child had fewer adjustment problems when more formal social support was also available to the family. The implications of these data for future research and for practice are discussed. PMID- 12757355 TI - Autism-related language, personality, and cognition in people with absolute pitch: results of a preliminary study. AB - Reports of a relatively high prevalence of absolute pitch (AP) in autistic disorder suggest that AP is associated with some of the distinctive cognitive and social characteristics seen in autism spectrum disorders. Accordingly we examined cognition, personality, social behavior, and language in 13 musicians with strictly defined AP (APS) and 33 musician controls (MC) without AP using standardized interviews and tests previously applied to identify the broad autism phenotype seen in the relatives of autistic probands. These included the Pragmatic Rating Scale (PRS) (social aspects of language) the Personality Assessment Schedule (PAS) (rigidity, aloofness, anxiety/worry, hypersensitivity), and WAIS performance subtests (PIQ). On the basis of their behavior in the interviews, subjects were classified as socially eccentric, somewhat eccentric, or not eccentric. Forty-six percent of the APS, but only 15% of the MC, were classified as socially eccentric (p < .03). APS but not MC showed higher scores on block design than on the other PIQ tests (p < .06), a PIQ pattern seen in autism spectrum disorders. Although APS and MC did not differ significantly on other measures it is of note that APS mean scores on the PRS and PAS (5.69, 4.92) were almost twice as high as those for the MC (3.03, 2.45). Thus, musicians with AP show some of the personality, language, and cognitive features associated with autism. Piecemeal information processing, of which AP is an extreme and rare example, is characteristic of autism and may be associated as well with subclinical variants in language and behavior. We speculate that the gene or genes that underlie AP may be among the genes that contribute to autism. PMID- 12757354 TI - Long-term memory in high-functioning autism: controversy on episodic memory in autism reconsidered. AB - Two studies were conducted to examine the nature of verbal long-term memory (LTM) in people with autism. In Study 1, undergraduate students showed better LTM and more verbal associations for concrete than abstract nouns. Probability of recall of the nouns strongly correlated with the number of associations with those nouns. In Study 2, unlike controls, autistic subjects did not show superior recall of concrete over abstract nouns despite overall comparable performance. A highly significant correlation between probability of recall and associative value was found only in the controls. Furthermore, there was an unusual correlation between LTM performance and a nonverbal measure in the autistic group. The results were discussed in terms of the relation between episodic memory and semantic memory. PMID- 12757356 TI - Case report: Angelman syndrome in an individual with a small SMC(15) and paternal uniparental disomy: a case report with reference to the assessment of cognitive functioning and autistic symptomatology. AB - The case of a 15-year-old male with Angelman syndrome, paternal uniparental disomy of chromosome 15, and a small supernumerary marker chromosome is discussed. Assessment of cognitive functioning revealed an uneven profile of ability across different domains; in particular, receptive language ability was found to be superior to expressive language ability, whilst both gross and fine motor skills were found to be relatively well developed. Assessment using the Autism Diagnostic Observation Schedule showed very little evidence of autistic symptomatology. The patient showed an interest in social interaction and used a variety of methods to communicate, including some gestures and several single words. A clinical history revealed febrile convulsions during childhood but an absence of seizures in the previous 5 years. The patient was not hypopigmented, and height, weight, and head circumference were within the normal range for his age. The implications of these features are discussed in the context of previous work describing a milder phenotype in nondeletion cases of Angelman syndrome and work that has examined the prevalence of autism spectrum disorders amongst individuals with Angelman syndrome. PMID- 12757357 TI - Partial tetrasomy of chromosome 3q and mosaicism in a child with autism. AB - In this report we describe the case of an 11-year-old male with autism and mental retardation, presenting a tetrasomy of chromosome 3q. Cytogenetic analysis showed a mosaic for an unbalanced karyotype consisting of mos46,XY,add(12)(p13.3)(56)/46,XY(45). FISH using WCP and subtelomeric probes identified the extra material on 12p to be an inverted duplication of the distal segment of chromosome 3q. Anomalies in chromosome 3q have not been previously described in association with autism, although association with psychomotor delays and behavior problems has been frequently reported and are here further discussed. This chromosomal 3q segment is therefore likely to include genes involved in specific neurodevelopment pathways, and further analysis of the region is warranted for the identification of the molecular alterations that lead to the autistic features described. PMID- 12757358 TI - The yield of the medical evaluation of children with pervasive developmental disorders. AB - Little information is available regarding the yield of the medical evaluation of children diagnosed with pervasive developmental disorder-not otherwise specified (PDD-NOS) compared to children diagnosed with autistic disorder. Medical records were reviewed for 182 patients less than 18 years of age with either PDD-NOS or autistic disorder evaluated between 1994 and 1998 at Mayo Clinic. A condition likely to be etiologically relevant was identified in 6/117 (5.1%) patients diagnosed with PDD-NOS and 2/65 (3.1%) patients diagnosed with autistic disorder. Genetic disorders, both chromosomal and single-gene, were the most commonly identified conditions. Seizure disorders, electroencephalogram abnormalities, and anomalies on brain imaging were common in both groups. The likelihood of uncovering an etiologically relevant condition in children diagnosed with either PDD-NOS or autistic disorder may be equivalent. The scope of the etiological search in an individual patient with an autistic spectrum disorder should not be limited by the specific diagnostic category. PMID- 12757360 TI - Autism and phenylketonuria. AB - Phenylketonuria (PKU) has been also reported in children with infantile autism (IA); however, the frequency of this association is variably reported. Patients with various forms of hyperphenylalaninemia (HPA) were evaluated applying two methods: the Autism Diagnostic Interview-Revised (ADI-R) and the Childhood Autism Rating Scale (CARS). A total of 243 patients were investigated, 97 with classical PKU, 62 identified by neonatal screening, and 35 late diagnosed. None out of 62 patients with classic PKU diagnosed early met criteria for autism. In the group of 35 patients diagnosed late, two boys (5.71%) ages 16 and 13 years fulfilled the diagnostic criteria for autism. The present study confirms that classical PKU is one of the causes of autism, but the prevalence seems to be very low. PMID- 12757359 TI - Two children with muscular dystrophies ascertained due to referral for diagnosis of autism. AB - We report two children who were referred for diagnostic assessment for autism and were subsequently determined to have a muscular dystrophy (MD). Each child had a history of speech delay and social impairments, but also had motor delays that had not previously been investigated. Both children met diagnostic criteria for autism spectrum disorders on standardized assessment. Each child was hypotonic and had other mild motor impairments. Serum creatine kinase (CK) activity was markedly elevated in each child, and subsequent muscle biopsy led to diagnosis of Becker's MD and congenital (autosomal recessive) MD, respectively. These cases highlight the importance of a thorough neuromotor examination for all children with suspected autism spectrum disorders. PMID- 12757361 TI - Brief report: pitocin induction in autistic and nonautistic individuals. AB - Oxytocin plays an important role in social-affiliative behaviors. It has been proposed that exposure to high levels of exogenous oxytocin at birth, via pitocin induction of delivery, might increase susceptibility to autism by causing a downregulation of oxytocin receptors in the developing brain. This study examined the rates of labor induction using pitocin in children with autism and matched controls with either typical development or mental retardation. Birth histories of 41 boys meeting the criteria for autistic disorder were compared to 25 age- and IQ-matched boys without autism (15 typically developing and 10 with mental retardation). There were no differences in pitocin induction rates as a function of either diagnostic group (autism vs. control) or IQ level (average vs. subaverage range), failing to support an association between exogenous exposure to oxytocin and neurodevelopmental abnormalities. PMID- 12757362 TI - Brief report: cognitive correlates of enlarged head circumference in children with autism. AB - This study examined the frequency and cognitive correlates of enlarged head circumference in a sample of 63 children with autism between the ages of 4 and 14. Consistent with prior evidence, macrocephaly occurred at a significantly higher frequency than in a normal reference sample. Head circumference was not associated with language or executive functioning, nor was it related to verbal or nonverbal IQ. Head circumference was, however, correlated with discrepancies between verbal and nonverbal IQ scores, independent of absolute level of verbal ability. Children with discrepantly high nonverbal abilities had a mean standardized head circumference that was more than 1 SD greater than in the reference sample, and that was significantly greater than in autistic children with a relative verbal advantage or no discrepancy in cognitive abilities, for whom mean head circumference was within normal limits. This convergence of physical and cognitive features suggests a possible etiologically significant subtype of autism. PMID- 12757363 TI - Developing an Estonian version of the Psychoeducational Profile Revised (PEP-R). PMID- 12757364 TI - Commentary: Bebko, Perry, and Bryson on Mostert (2001), "facilitated communication since 1995". PMID- 12757366 TI - Functional assessment. PMID- 12757365 TI - Commentary: Blaxill, Baskin, and Spitzer on Croen et al. (2002), the changing prevalence of autism in California. PMID- 12757367 TI - Transcortical alterations in Na(+)-K+ ATPase and microtubule-associated proteins immunoreactivity in the rat cortical atrophy model induced by hypoxic ischemia. AB - To identify the chronological transcortical change in the contralateral hemisphere following ischemic insults, we investigated the changes in microtubule associated protein (MAP) and Na(+)-K+ ATPase expressions in the peri-infarct zone and contralateral hemisphere, including the hippocampus. Two days after hypoxic ischemia, Na(+)-K+ ATPase immunoreactivity was significantly enhanced in the contralateral cortex and was maintained up to 7 days after ischemia, whereas Na(+)-K+ ATPase immunoreactivity in the peri- and infarct zones was unaffected by hypoxic ischemia. In contrast, 2 to 7 days after ischemia, MAP1A and MAP2 immunoreactivity in the ipsi- and contralateral cortex significantly decreased, whereas in layer V, MAP1 immunoreactivity obviously accumulated in the neurons and their processes. In the hippocampus, 2 days after insults both MAP1A and MAP2 immunoreactivity was significantly reduced within the ipsi- and contralateral hippocampus. In the contralateral hippocampus, however, the distribution of MAP2 immunoreactivity recovered to the sham level 7 days after ischemia, whereas MAP1A immunoreactive axons remained 2 months after ischemia. The results suggest that the unilateral elevation of Na(+)-K+ ATPase immunoreactivity reflects elevated neuronal activity. In addition, this asymmetric hyperexcitability might play an important role in the recovery or the reorganization of the brain, accompanied by transcortical changes in MAPs expression. PMID- 12757368 TI - Chronic restraint stress induces an isoform-specific regulation on the neural cell adhesion molecule in the hippocampus. AB - Existing evidence indicates that 21-days exposure of rats to restraint stress induces dendritic atrophy in pyramidal cells of the hippocampus. This phenomenon has been related to altered performance in hippocampal-dependent learning tasks. Prior studies have shown that hippocampal expression of cell adhesion molecules is modified by such stress treatment, with the neural cell adhesion molecule (NCAM) decreasing and L1 increasing, their expression, at both the mRNA and protein levels. Given that NCAM comprises several isoforms, we investigated here whether chronic stress might differentially affect the expression of the three major isoforms (NCAM-120, NCAM-140, NCAM-180) in the hippocampus. In addition, as glucocorticoids have been implicated in the deleterious effects induced by chronic stress, we also evaluated plasma corticosterone levels and the hippocampal expression of the corticosteroid mineralocorticoid receptor (MR) and glucocorticoid receptor (GR). The results showed that the protein concentration of the NCAM-140 isoform decreased in the hippocampus of stressed rats. This effect was isoform-specific, because NCAM-120 and NCAM-180 levels were not significantly modified. In addition, whereas basal levels of plasma corticosterone tended to be increased, MR and GR concentrations were not significantly altered. Although possible changes in NCAM-120, NCAM-180 and corticosteroid receptors at earlier time points of the stress period cannot be ignored, this study suggests that a down-regulation of NCAM-140 might be implicated in the structural alterations consistently shown to be induced in the hippocampus by chronic stress exposure. As NCAM-140 is involved in cell-cell adhesion and neurite outgrowth, these findings suggest that this molecule might be one of the molecular mechanisms involved in the complex interactions among neurodegeneration-related events. PMID- 12757369 TI - Different modes of pitch perception and learning-induced neuronal plasticity of the human auditory cortex. AB - We designed a melody perception experiment involving eight harmonic complex tones of missing fundamental frequencies (hidden auditory object) to study the short term neuronal plasticity of the auditory cortex. In this experiment, the fundamental frequencies of the complex tones followed the beginning of the virtual melody of the tune "Frere Jacques". The harmonics of the complex tones were chosen so that the spectral melody had an inverse contour when compared with the virtual one. Evoked magnetic fields were recorded contralaterally to the ear of stimulation from both hemispheres. After a base line measurement, the subjects were exposed repeatedly to the experimental stimuli for 1 hour a day. All subjects reported a sudden change in the perceived melody, indicating possible reorganization of the cortical processes involved in the virtual pitch formation. After this switch in perception, a second measurement was performed. Cortical sources of the evoked gamma-band activity were significantly stronger and located more medially after a switch in perception. Independent Component Analysis revealed enhanced synchronization in the gamma-band frequency range. Comparing the gamma-band activation of both hemispheres, no laterality effects were observed. The results indicate that the primary auditory cortices are involved in the process of virtual pitch perception and that their function is modifiable by laboratory manipulation. PMID- 12757370 TI - Correlation of clinical neuromusculoskeletal and central somatosensory performance: variability in controls and patients with severe and mild focal hand dystonia. AB - Focal hand dystonia (FHd) is a recalcitrant, disabling movement disorder, characterized by involuntary co-contractions of agonists and antagonists, that can develop in patients who overuse or misuse their hands. The aim of this study was to document clinical neuromusculoskeletal performance and somatosensory responses (magnetoencephalography) in healthy controls and in FHd subjects with mild versus severe hand dystonia. The performance of healthy subjects (n = 17) was significantly better than that of FHd subjects (n =17) on all clinical parameters. Those with mild dystonia (n = 10) demonstrated better musculoskeletal skills, task-specific motor performance, and sensory discrimination, but the performance of sensory and fine motor tasks was slower than that of patients with severe dystonia. In terms of somatosensory evoked field responses (SEFs), FHd subjects demonstrated a significant difference in the location of the hand representation on the x and y axes, lower amplitude of SEFs integrated across latency, and a higher ratio of mean SEF amplitude to latency than the controls. Bilaterally, those with FHd (mild and severe) lacked progressive sequencing of the digits from inferior to superior. On the affected digits, subjects with severe dystonia had a significantly higher ratio of SEF amplitude to latency and a significantly smaller mean volume of the cortical hand representation than those with mild dystonia. Severity of dystonia positively correlated with the ratio of SEF mean amplitude to latency (0.9029 affected, 0.8477 unaffected; p < 0.01). The results of the present study strengthen the evidence that patients with FHd demonstrate signs of somatosensory degradation of the hand that correlates with clinical sensorimotor dysfunction, with characteristics of the de differentiation varying by the severity of hand dystonia. If these findings represent aberrant learning, then effective rehabilitation must incorporate the principles of neuroplasticity. Training must be individualized to each patient to rebalance the sensorimotor feedback loop and to restore normal fine motor control. PMID- 12757371 TI - Experience-dependent color constancy in guppies (Poecilia reticulata). AB - We investigated the ability to recognize the color of surfaces in fish (Poecilia reticulata), bred from birth in conditions of artificial light with constant spectral content. The capacity for color constancy significantly deteriorated when compared that to the control group. Further alteration of lighting conditions and transfer into natural daylight conditions restored the suppressed function to its normal level. We suggest that the color constancy function belongs in the visual system-response functions, the full development of which requires the accumulation of individual visual experience. PMID- 12757372 TI - A rapid method for the construction of oligonucleotide arrays. AB - A simple method has been devised to construct oligonucleotide array on a variety of surfaces using commonly available reagents and chemistry with good efficiency and accuracy. The method involves the generation of hydroxyl functionalities on glass, polypropylene, polyethylene, and commonly used surfaces for construction of oligonucleotide arrays followed by their activation with trifluoroethanesulfonyl chloride (tresyl chloride). The activated surface in the subsequent reaction is used to covalently immobilize oligonucleotides in regioselective fashion to create an oligonucleotide array. The surface bound tresyl sulfonate esters allow the immobilization of oligonucleotides specifically via their 3'- or 5'-end having mercaptohexyl- or aminohexyl functionalities. The constructed oligonucleotide arrays were successfully used to analyze oligonucleotides by hybridization technique. PMID- 12757373 TI - Photoenhancement of transfection efficiency using novel cationic lipids having a photocleavable spacer. AB - New cationic lipids having an o-nitrobenzyl moiety as a photocleavable spacer between its hydrophilic and hydrophobic region were synthesized. To improve the efficiency of transfection with lipoplexes, after transfecting the cationic lipid aggregate/DNA complex, photoirradiation was performed. Photochemical decomposition of lipids would not only make the vector's membrane unstable to facilitate the fusion with endocytic vesicles, but also promote dissociation of cationic lipid-DNA complex, thus aiding the escape of DNA from the endocytic vesicles. Using a luciferase gene as a model, we show that UV irradiation of photoresponsive lipoplex-treated COS-1 cells induces a substantial increase in the efficiency of transfection. Herein, we show a novel photoresponsive gene delivery system. PMID- 12757374 TI - Temperature-induced switching of enzyme activity with smart polymer-enzyme conjugates. AB - A method for thermally induced switching of enzyme activity has been developed, based on the site-directed conjugation of end-reactive temperature-responsive polymers to a unique cysteine (Cys) residue positioned near the enzyme active site. The reversible temperature-induced collapse of N,N-dimethylacrylamide (DMA)/N-4-phenylazo-phenylacrylamide (AZAAm) copolymers (DMAAm) has been used as a molecular switch to control the catalytic activity of endoglucanase 12A (EG 12A). The polymer was conjugated to the EG 12A site-directed mutant N55C, directly adjacent to the cellulose binding cleft, and to the S25C mutant, where the conjugation site is more distant. The N55C conjugate displayed a larger activity shutoff efficiency in the collapsed polymer state than the S25C conjugate. Increasing the polymer molecular weight was also shown to increase the shutoff efficiency of the switch. Related to these effects of conjugation site and polymer size, the switching efficiency was found to be strongly dependent on substrate size. With a small substrate, o-nitrophenyl-beta-d-cellobioside (ONPC), there was minimal blocking of enzyme activity when the polymer was in the expanded state. With a large substrate, hydroxyethyl cellulose (HEC), there was a large reduction of enzyme activity in the polymer expanded state, even with relatively small polymer chains, and a further reduction when the polymer was collapsed. Similar general trends for the interactive effects of conjugation site, polymer size, and substrate size were observed for immobilized conjugates. Kinetic studies demonstrated that the switching activity was due to the blocking of substrate association by the collapsed polymers. These investigations provide mechanistic insight that can be utilized to design molecular switches for a variety of stimuli-responsive polymer-protein conjugates. PMID- 12757375 TI - Synthesis of polymerized thin films for immobilized ligand display in proteomic analysis. AB - We describe a new material for the display of biomolecular ligands for use in proteomic analysis. We report here on the construction of the first functionalized polymerized diacetylene thin films (PDTFs) for use in displaying immobilized ligands and their application in mass spectral proteomic analysis. Functionalized polymerized thin film surfaces were constructed with diacetylene containing biotin lipid monomers designed for the capture of proteins (streptavidin) from a complex cellular lysate and detection with mass spectrometry (MS). These materials serve as a prototype for ligand-based spotted arrays amenable to high throughput screening. Functionalized PDTFs can be easily manufactured for customized microarrays and demonstrate high protein specificity and low nonspecific protein adsorption, and the resulting microarrays constructed from these materials are compatible with several different protein analysis platforms. Our results suggest that these materials have broad potential applications for use in mass spectral-based proteomic analysis. PMID- 12757376 TI - A concise method for the preparation of peptide and arginine-rich peptide conjugated antisense oligonucleotide. AB - Peptide-oligonucleotide conjugates were synthesized using two strategies: a mimetic signal peptide-conjugated oligonucleotide was assembled stepwise on CPG by using 2,2-dimethyl-3-hydroxypropionic acid as a linker. To solve the precipitation problem in the coupling reaction caused by the electrostatic interaction of arginine-rich peptides and oligonucleotide, oligonucleotides were absorbed on an anion-exchange resin, and then the on-resin fragment was applied for the conjugation with arginine-rich peptide. The peptide-antisense oligonucleotides showed permeability to the cell membrane of HepG-2 cells. PMID- 12757377 TI - Enhanced tumor detection using a folate receptor-targeted near-infrared fluorochrome conjugate. AB - Fluorescence optical imaging technologies are currently being developed to image specific molecular targets in vivo. Detection technologies range from those providing microscopic detail to whole body imaging systems with potential clinical use. A number of target-specific near-infrared imaging probes have recently been developed to image receptors, antigens, and enzymes. The goal of the current study was to evaluate a new near-infrared (NIR) folate receptor (FR) targeted imaging probe for its ability to improve detection of FR-positive cancers. We hypothesized that modification of folate would retain receptor affinity in vivo, despite the bulkier NIR fluorochrome, NIR2 (em = 682 nm). Cellular uptake of the NIR conjugates was significantly higher in FR-positive nasopharyngeal epidermoid carcinoma, KB cells, compared to FR-negative human fibrosarcoma, HT1080 cells. When tumors were implanted in vivo, equal-sized KB tumors showed a 2.4-fold higher signal intensity compared to HT1080 tumors (24 h). The maximum signal-to-background ratio (3-fold) was observed at 24 h in KB tumor. Injection of the unmodified NIR2 fluorochrome did not result in persistent contrast increases under similar conditions. Furthermore, tumor enhancement with the NIR2-folate probe persisted over 48 h and was inhibitable in vivo by administration of unlabeled folate. These results indicate that folate-modified NIR fluorochrome conjugate can be used for improved detection of FR-positive tumors. PMID- 12757378 TI - Monoclonal antibody radiopharmaceuticals: cationization, pegylation, radiometal chelation, pharmacokinetics, and tumor imaging. AB - The 528 murine monoclonal antibody (MAb) to the human epidermal growth factor receptor (EGFR) was sequentially cationized with hexamethylenediamine and conjugated with diethylenetriaminepentaacetic acid (DTPA) as a potential antibody radiopharmaceutical for imaging EGFR-expressing cancer. The cationized 528 MAb was characterized with isoelectric focusing and electrophoresis, and an immunoradiometric assay, which showed the affinity of the 528 MAb for the human EGFR was retained following cationization. The native or cationized 528 MAb, labeled with (111)In, was injected intravenously in scid mice bearing human U87 flank tumors, which express the EGFR, and tumor imaging was performed with both external detection in live animals and with whole body autoradiography. However, the tumor signal was not increased with the cationized MAb, relative to the native MAb, and this was due to a serum inhibition phenomenon that was confirmed by a pharmacokinetics analysis in control mice. In an attempt to block the serum inhibition, the cationized 528 MAb was pegylated with 2000 Da poly(ethylene glycol), and the cationized/pegylated MAb was conjugated with DTPA and labeled with (111)In. However, a pharmacokinetics analysis showed the pegylation did not reverse the serum inhibition of the cationic charge on the MAb. These studies describe methods for reformulating monoclonal antibodies to develop improved radiopharmaceuticals, but show that radiolabeling a cationized MAb with DTPA produces a serum neutralization of the initial cationization modification. PMID- 12757379 TI - A cysteine-linkable, short cleavable photoprobe with dual functionality to explore protein-protein interfaces. AB - We developed a bifunctional photoprobe with dual functionality, that can be specifically tethered to cysteinyl residues of peptides and proteins through a short cleavable disulfide bond. Thus, an aryldiazonium moiety is positioned at approximately 8.5 A from the modified cysteinyl alpha-carbon, leading to one of the shortest cleavable linkages. In a sodium azide-containing buffer, the aryldiazonium moiety is transformed into an aryl azide. Therefore, with one bifunctional photoprobe two types of photogenerated species can be obtained: a hydrophilic and positively charged arylcation or a hydrophobic nitrene. We coupled the aryldiazonium probe, in a site-directed manner, to a nicotinic acetylcholine receptor competitive antagonist, obtained by chemical engineering of an analogue of a snake alpha-neurotoxin. In this molecule, Arg33, which is known to interact with the receptor, was replaced by a cysteine residue, where the photoprobe could be attached. Under inactinic light, this novel photosensitive snake toxin behaved as a reversible ligand on the Torpedo acetylcholine receptor. However, when irradiated at 391 nm, it generated a highly reactive arylcation which labeled mostly the receptor alpha-subunit, confirming the location of the tip of the second toxic loop near this receptor subunit. Finally, we showed that reduction of the disulfide bond, linking the ligand to the photocoupled receptor, allowed introduction of radioactivity on the labeled residue(s), opening the way to further characterization and avoiding the synthesis of a radioactive bifunctional photoprobe. PMID- 12757380 TI - Folate-targeted PEG as a potential carrier for carboplatin analogs. Synthesis and in vitro studies. AB - Like most low molecular weight drugs, carboplatin has a short blood circulation time, which reduces tumor uptake and intracellular DNA binding. Drugs conjugated to PEG carriers benefit from prolonged blood circulation, but suffer from reduced cell permeability. In this work we attempted to develop long-circulating PEGylated carboplatin analogues with improved cell permeation abilities, by conjugating the platinum moiety to folate-targeted PEG carriers capable of utilizing the folate receptor-mediated endocytosis (FRME). Two bifunctional FA PEG conjugates, FA-PEG-Pt and FA-PEG-FITC, were prepared, and their cell uptake, DNA binding, and cytotoxicity were studied by fluorescent microscopy, FACS, and platinum analysis. Folate-targeted PEG conjugates enter the cells efficiently by the FRME pathway but form relatively few DNA adducts and have higher IC(50) values than carboplatin and their nontargeted analogues. Nontargeted PEG-Pt conjugates have a lower cellular uptake but produce higher levels of DNA binding and improved cytotoxicity. Carboplatin, used as a control, has the fastest cellular uptake, but after 16 h of postincubation a large percentage of the drug is excreted from the cells. The findings of this study suggest that folate targeted conjugates such as FA-PEG-Pt, may not be an optimal prodrug for the carboplatin family compounds, because the conjugates or the active moieties are neutralized or blocked during the FRME process and do not manage to effectively reach the nuclear DNA. PMID- 12757381 TI - Affinity thermoprecipitation and recovery of biotinylated biomolecules via a mutant streptavidin-smart polymer conjugate. AB - A system has been developed for reversibly binding and thermoprecipitating biotinylated macromolecules. A high off-rate Ser45Ala (S45A) streptavidin mutant has been covalently conjugated to poly(N-isopropylacrylamide) (PNIPAAm), a temperature-responsive polymer. The resulting conjugate is shown to coprecipitate biotinylated immunoglobulin G (IgG) and a biotinylated oligonucleotide in response to a thermal stimulus. Thermally precipitated biotinylated macromolecules can be released from the S45A-PNIPAAm conjugate by simple treatment with excess free biotin. This release step has been shown to be unique to the mutant streptavidin conjugate-a conjugate of wild type (WT) streptavidin and PNIPAAm does not release bound biotinylated molecules upon treatment with excess free biotin. The capture efficiency (fraction of target molecule precipitated from solution) of the S45A-PNIPAAm conjugate is similar to that of the WT-PNIPAAm conjugate for the biotinylated IgG target molecule (near 100%), but significantly smaller for the biotinylated oligonucleotide target (approximately 60% for the S45A-PNIPAAm conjugate compared to 80% for the WT PNIPAAm conjugate). The release efficiency (fraction of originally precipitated target molecule released after treatment with free biotin) of the S45A-PNIPAAm conjugate is 70-80% for the biotinylated IgG target and nears 100% for the biotinylated oligonucleotide target. This system demonstrates the use of a high off-rate streptavidin mutant to add reversibility to a system based on smart polymer-streptavidin conjugates. PMID- 12757382 TI - Synthesis of linear polyethylenimine derivatives for DNA transfection. AB - A series of linear polymers containing varying amounts of ethylenimine or N propylethylenimine units were synthesized by hydrolysis and/or reduction of polyethyloxazolines. The pK(a)s of the polyamines were determined potentiometrically. Gel mobility shift assay showed that the efficiency of DNA complexation was related to the fraction of amino groups that are protonated at neutral pH. The effects of cationic charge density and molar weight of the polymers on the transfection efficiency were evaluated on HepG2 cells. The results obtained with different copolymers show that the transfection efficiency primarily depends on the fraction of ethylenimine units included in the polymer albeit the molar weight is also of importance. On the basis of the results obtained with poly(N-propylethylenimines), we also demonstrate that the high transfection efficiency of polyethylenimines does not solely rely on their capacity to capture protons which are transferred into the endo-lysosomes during acidification. PMID- 12757383 TI - Synthesis and properties of ester-linked peptide nucleic acid prodrug conjugates. AB - A Boc-protected amino acid containing an ester function, 2-([N-Boc glycyl]oxymethyl)benzoic acid, has been synthesized and incorporated into peptide nucleic acid (PNA) oligomers. In model experiments it is found that the ester is fairly stable in aqueous solution at pH 7.4 and 37 degrees C (t(1/2) = 6 h), whereas it is rapidly cleaved in mouse serum and in kidney and liver homogenates (t(1/2) = 0.1-0.5 min). Furthermore, ester-linked fatty acid PNA conjugates targeted to an aberrant splice site in luciferase mRNA were prepared and shown to be twice as potent for inducing active luciferase as the corresponding conjugate not containing the linker. Thus, a PNA prodrug approach may be useful for both ex vivo as well as in vivo applications. PMID- 12757384 TI - Atomic force microscopy of liposomes bearing fibrinogen. AB - Extruded liposomes formed from dipalmitoylphosphatidylcholine and cholesterol, with and without fibrinogen, were examined by atomic force microscopy (AFM). The sequence of events involved in the transition from attached liposomes to bilayer patches on mica supports was viewed by tapping mode in liquid. After adhesion to the mica surface, both liposomes without fibrinogen and liposomes with attached fibrinogen collapsed into patches. The fibrinogen layer attached to the liposomes was 2.6 nm thick. This implied that the protein was spread over the entire liposome and the protein characteristic trinodular structure disappeared. To check the type of bond between fibrinogen and liposome, sequential images were taken after the incubation of fibrinogen with liposomes with and without a chemical group for attaching the protein. The results clearly confirmed that fibrinogen bound covalently to liposomes. PMID- 12757385 TI - Interaction of a ferrocenoyl-modified peptide with papain: toward protein sensitive electrochemical probes. AB - A new ferrocenoyl tetrapeptide, Fc-Gly-Gly-Tyr-Arg-OH, has been synthesized, which acts as an effective competitive inhibitor to papain, with a K(i) of 9 microM at pH 6.2. The electrochemical potential of the ferrocenoyl moiety is influenced by papain binding, resulting in a small cathodic shift of 30 mV. PMID- 12757386 TI - Nucleosides and nucleotides. 218. Alternate-strand triple-helix formation by the 3'-3'-linked oligodeoxynucleotides using a purine motif. AB - In this paper, we describe the synthesis of the 3'-3'-linked TFOs that can form the antiparallel triplexes with the duplex DNA target by reverse Hoogsteen hydrogen bonds. Stability of the alternate-strand triplexes between these TFOs and the target DNAs was investigated using the electrophoretic mobility shift assay (EMSA). It was found that the alternate-strand triplexes were significantly stabilized by linking the TFO fragments with the pentaerythritol linker. And, unlike the alternate-strand triplexes composed of the pyrimidine motif, the terminal ammonium ion of the aminobutyl-linker and the intercalator of the TFOs did not contribute to the stability of the alternate-strand triplex comprised of the purine motif. We also tested the ability of the 3'-3'-linked TFOs to inhibit cleavage of the duplex DNA target 17 by the restriction enzyme EcoT14I and found that the 3'-3'-linked TFOs 12 and 13 inhibited the cleavage by the enzyme more effectively than the unlinked decamer 8. Thus, the TFOs linked with pentaerythritol may be useful as the antigene oligonucleotide to the DNA targets, which have alternating oligopyrimidine-oligopurine sequences. PMID- 12757387 TI - A novel method for the rational construction of well-defined immunogens: the use of oximation to conjugate cholera toxin B subunit to a peptide-polyoxime complex. AB - Cholera toxin B subunit (CTB), capable of binding to all mucous membranes in its pentameric form, is a potential carrier of mucosal vaccines. In our previous work we reported that the N-terminus of CTB, a threonine, could in principle undergo oxidation and oximation to form conjugates with a cascade of immunogenic peptides. In this study, we set up a model by chemically coupling CTB to a polyoxime that possessed five copies of influenza virus-derived peptides displayed in comblike form. The construct was reconstituted into pentameric form when eluted from a Superdex column after conjugation, and the pentameric nature of this CTB-viral peptide complex was confirmed by SDS-PAGE. GM(1)-ELISA assay showed that the binding properties of CTB-viral peptide complex were increased 4 5-fold over native CTB. PMID- 12757388 TI - Site-specific quantitative evaluation of the protein glycation product N6-(2,3 dihydroxy-5,6-dioxohexyl)-L-lysinate by LC-(ESI)MS peptide mapping: evidence for its key role in AGE formation. AB - Advanced glycation end products (AGEs) contribute to various pathologies associated with the general aging process and long-term complications of diabetes. Involvement of alpha-dicarbonyl intermediates in the formation of such compounds is firmly established. We now report on the first unequivocal identification of the dideoxyosone N(6)-(2,3-dihydroxy-5,6-dioxohexyl)-l-lysinate (4) on lysozyme via its quinoxaline derivative N(6)-(2,3-dihydroxy-4-quinoxalin-2 ylbutyl)-l-lysinate (6), formed by reaction of 4 with o-phenylenediamine (OPD). For accurate quantification of the total content of 6 as well as of glucosepane 5 by LC-(ESI)MS, (13)C(6)-labeled reference compounds were independently synthesized; 5 so far is the only established follow-up product of 4. With an overall lysine derivatization quota of 5%, compound 4 is shown to be a quantitatively important Maillard intermediate of which only about 8 per thousand are transformed into the cross-link 5. Hence, the major follow-up products of the highly reactive intermediate 4 are yet unknown. The site-specific quantitative evaluation of aminoketose 1 and quinoxaline 6 by LC-(ESI)MS peptide mapping shows that all lysine moieties in lysozyme are in fact modified by these compounds. If an arginine side chain is adjacent to the lysine moiety, transformation of 1 into 4 seems to be favored. The efficient formation and high reactivity of 4 clearly points to its potential as exogenous or endogenous glycotoxin. PMID- 12757389 TI - Synthesis and biological evaluation of halogenated naphthyridone carboxamides as potential ligands for in vivo imaging studies of substance P receptors. AB - With the aim of developing new radioligands for in vivo studies of substance P receptors using positron emission tomography or single photon emission computed tomography, 2- and 3-halo naphthyridone-6-carboxamide derivatives were synthesized. Their affinities toward the target receptors were evaluated on CHO cells and compared to the unsubstituted analogue EP 00652218 (IC(50) = 100 nM +/- 20). The IC(50) value was not altered in the case of 2-chloro compound 1 (IC(50) = 100 nM +/- 15) and only slightly reduced for the 2-fluoro and -iodo analogues 6 and 8 (IC(50) = 500 nM +/- 80). A drastic reduction in binding (IC(50) > 1000 nM) was observed for the halogenated compounds 2-5, 7, and 9. PMID- 12757390 TI - New reagents for the introduction of reactive functional groups into chemically synthesized DNA probes. AB - An efficient and versatile preparative approach is described, allowing for the preparation of DNA probes modified with an aldehyde group at the 3'- or 5'-end. The developed synthetic strategy allows for the preparation of a new family of phosphoramidites and solid supports compatible with the automated synthesis of modified oligonucleotide probes. These new reagents were prepared from intermediates 3 and 3a, obtained from the commercially available aleuritic acid 1. It was demonstrated that the new phosphoramidite reagents also could be used as new types of cleavable linkers. A new and efficient method for the production of 5' aldehyde-labeled DNA probes was developed. PMID- 12757391 TI - Extending the applicability of carboxyfluorescein in solid-phase synthesis. AB - Optimized coupling protocols are presented for the efficient and automated generation of carboxyfluorescein-labeled peptides. Side products, generated when applying earlier protocols for the in situ activation of carboxyfluorescein, were eliminated by a simple procedure, yielding highly pure fluorescent peptides and minimizing postsynthesis workup. For the cost-efficient labeling of large compound collections, coupling protocols were developed reducing the amount of coupling reagent and fluorophore. To enable further chemical derivatization of carboxyfluorescein-labeled peptides in solid-phase synthesis, the on-resin introduction of the trityl group was devised as a protecting group strategy for carboxyfluorescein. This protecting group strategy was exploited for the synthesis of peptides labeled with two different fluorescent dyes, essential tools for bioanalytical applications based on fluorescence resonance energy transfer (FRET). Tritylation and optimized labeling conditions led to the development of a fluorescein-preloaded resin for the automated synthesis of fluorescein-labeled compound collections with uniform labeling yields. PMID- 12757393 TI - Enhanced hydrolytic stability and water solubility of an aromatic nitrogen mustard by conjugation with molecular umbrellas. AB - Chlorambucil, an aromatic nitrogen mustard, has been conjugated to putrescine- and spermidine-based scaffolds bearing one, two, and four persulfated cholic acid units. Those conjugates bearing two or four sterols show improved hydrolytic stability and water solubility relative to chlorambucil. A similar conjugate that contained only one sterol unit shows negligible improvement in hydrolytic stability but a significant increase in water solubility. Qualitatively, the hydrolytic stability within this series of conjugates parallels the shielding effects that have previously been found for related conjugates bearing a pendant, hydrophobic fluorescent probe. In vitro studies indicate that these conjugates possess modest to moderate activity against certain human lymphoblastic leukemia and human colon carcinoma cells. PMID- 12757392 TI - Utility of poly(ethylene glycol) conjugation to create prodrugs of amphotericin B. AB - This paper reports on the synthesis, safety, and efficacy of a series of water soluble derivatives of poly(ethylene glycol) (PEG)-conjugated amphotericin B (AmB). PEG 40 000 attached to the sugar amino group of AmB via labile carbamate and carbonate linkages was examined. The synthetic program conducted for this investigation provided a series of disubstituted PEG-AmB derivatives which had in vitro PEG half-life of hydrolyses rates in rat plasma varying between 1 and 3 h. Importantly, all conjugates demonstrated less than 6% hydrolysis following 24 h incubation in pH 7.4 phosphate buffer at 25 degrees C and showed solubilities greater than 46 mg/mL in aqueous solutions. The solubility of AmB in the conjugates increased up to approximately 200 times compared to unmodified AmB in saline. As a major finding, this investigation demonstrated that conjugation of PEG to AmB could produce conjugates that were significantly (6x) less toxic than AmB-deoxycholate and maintained, or even had enhanced, in vivo antifungal activity. PMID- 12757394 TI - Structural effects of carbohydrate-containing polycations on gene delivery. 3. Cyclodextrin type and functionalization. AB - Linear cationic beta-cyclodextrin (beta-CD)-based polymers can form polyplexes with plasmid DNA and transfect cultured cells. The effectiveness of the gene delivery and the cellular toxicity has been related to structural features in these polycations. Previous beta-CD polycations were prepared from the cocondensation of 6(A),6(D)-dideoxy-6(A),6(D)-diamino-beta-CD monomers with other difunctionalized monomers such as dimethyl suberimidate (DMS). Here, the type of CD and its functionalization are varied by synthesizing numerous 3(A),3(B) dideoxy-3(A),3(B)-diamino-beta- and gamma-CD monomers. Both alkyl- and alkoxydiamines are prepared in order to vary the nature of the spacing between the CD and the primary amines in the monomers. These diamino-CD-monomers are polymerized with DMS to yield amidine-based polycations. The nature of the spacer between the CD-ring and the primary amines of each monomer is found to influence both molecular weight and polydispersity of the polycations. When these polycations are used to form polyplexes with plasmid DNA, longer alkyl regions between the CD and the charge centers in the polycation backbone increase transfection efficiency and toxicity in BHK-21 cells, while increasing hydrophilicity of the spacer (alkoxy versus alkyl) provides for lower toxicity. Further, gamma-CD-based polycations are shown to be less toxic than otherwise identical beta-CD-based polycations. PMID- 12757395 TI - PNA-based RNA-triggered drug-releasing system. AB - A three-component sequence-specific RNA-triggered drug-releasing system is described that consists of an 8-mer PNA linked to a coumarin ester (the prodrug component) and a 14-mer PNA linked to histidine (the catalytic component) that are complementary to the C loop of E. coli 5S rRNA (the triggering component). Binding of the catalytic component to the RNA creates a prodrug-metabolizing enzyme that catalyzes a 60,000-fold acceleration in the rate of coumarin release from the prodrug compared to the rate of coumarin release from the ester subunit catalyzed by imidazole alone. RNA-triggered release of hydroxycoumarin is only slightly less efficient than that triggered by a short unfolded DNA sequence corresponding to the PNA binding sites. The lower efficiency results from a decrease in k(cat) and an increase in K(M), presumably due to the bent nature of the RNA. The efficiency of DNA-triggered hydroxycoumarin release was found to depend on the distance between the catalytic and prodrug components. PMID- 12757396 TI - Synthesis of 3'-3'-linked oligonucleotides branched by a pentaerythritol linker and the thermal stabilities of the triplexes with single-stranded DNA or RNA. AB - Synthesis of 3'-3'-linked oligonucleotides branched by a pentaerythritol linker is described. The branched oligonucleotides were synthesized on a DNA/RNA synthesizer using a controlled pore glass (CPG) with a pentaerythritol linker carrying 4,4'-dimethoxytrityl (DMTr) and levulinyl (Lev) groups. The stability of the triplexes between the branched oligonucleotides and the target single stranded DNA or RNA was studied by thermal denaturation. The oligonucleotides with the pentaerythritol linker formed thermally stable triplexes with the single stranded DNA and RNA. Furthermore, the branched oligonucleotides containing 2'-O methylribonucleosides, especially the oligonucleotide composed of 2' deoxyribonucleosides and 2'-O-methylribonucleosides, stabilized the triplexes with the single-stranded DNA or RNA. Thus, the branched oligonucleotide containing 2'-O-methylribonucleosides may be a candidate for a novel antisense molecule by the triplex formation. PMID- 12757397 TI - Synthesis of antisense oligonucleotides carrying modified 2-5A molecules at their 5'-termini and their properties. AB - The synthesis of 8-methyladenosine-substituted 2-5A tetramers with hydroxyalkyl groups at the 5'-phosphates and the corresponding 2-5A-antisense chimeras is described. These oligonucleotides were synthesized by the phosphoramidite method with a DNA/RNA synthesizer. These 2-5A tetramers with hydroxyethyl and hydroxybutyl groups at their 5'-phosphates were more resistant to hydrolysis by alkaline phosphatase than those without the hydroxyalkyl groups. Incorporation of the hydroxyethyl group into the 2-5A tetramer and 2-5A-antisense chimera slightly reduced the abilities of their analogues to activate recombinant human RNase L, but the abilities of the 2-5A tetramer and the 2-5A-antisense chimera both with the hydroxyethyl group and 8-methyladenosine returned to 80 and 50% relative to those of the oligonucleotides without the hydroxyethyl group and 8 methyladenosine, respectively. Furthermore, the enzyme activated by 8 methyladenosine-substituted 2-5A-antisense chimera with the hydroxyethyl group cleaved the complementary RNA as efficiently as that activated by 2-5A-antisense chimera without the hydroxyethyl group and 8-methyladenosine. Thus, the 2-5A antisense chimera carrying the hydroxyethyl group and 8-methyladenosine will be a candidate for a novel antisense molecule. PMID- 12757398 TI - Site-specific fluorescent labeling of DNA using Staudinger ligation. AB - We report the site-specific fluorescent labeling of DNA using Staudinger ligation with high efficiency and high selectivity. An oligonucleotide modified at its 5' end by an azido group was selectively reacted with 5-[(N-(3'-diphenylphosphinyl 4'-methoxycarbonyl)phenylcarbonyl)aminoacetamido]fluorescein (Fam) under aqueous conditions to produce a Fam-labeled oligonucleotide with a high yield (approximately 90%). The fluorescent oligonucleotide was characterized by matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Because of the relatively high yield of the Staudinger ligation, simple purification of the product by size-exclusion chromatography and desalting is sufficient for the resulting fluorescent oligonucleotide to be used as a primer in a Sanger dideoxy sequencing reaction to produce fluorescent DNA extension fragments, which are analyzed by a fluorescent electrophoresis DNA sequencer. The results indicate that the Staudinger ligation can be used successfully and site specifically to prepare fluorescent oligonucleotides to produce DNA sequencing products, which are detected with single base resolution in a capillary electrophoresis DNA sequencer using laser-induced fluorescence detection. PMID- 12757399 TI - Effects of levocarnitine on mitochondrial antioxidant systems and oxidative stress in aged rats. AB - OBJECTIVE: Levocarnitine is important in beta-oxidation of fatty acids. We evaluated the role of levocarnitine in the skeletal muscle mitochondrial antioxidant system of aged rats. METHODS: Male albino Wistar rats were used in this study. The animals were divided into two groups: young rats (group I) and aged rats (group II). These rats were further subdivided into three groups: one control group (groups Ia and IIa) and two experimental groups (groups Ib, IIb and Ic, IIc) for supplementation with levocarnitine for 14 and 21 days, respectively. After the experimental period, the animals were killed by cervical decapitation; blood and skeletal muscle were isolated for further analysis. RESULTS: Levels of antioxidant enzymes such as superoxide dismutase, catalase and glutathione peroxidase, and non-enzymatic antioxidants such as reduced glutathione, ascorbic acid (vitamin C) and tocopherol (vitamin E) were found to be decreased in the blood and skeletal muscle mitochondria of aged rats. Supplementation with levocarnitine in aged rats improved the antioxidant status in a dose-dependent manner. CONCLUSIONS: These findings demonstrated that levocarnitine enhances the activity of the mitochondrial antioxidant system and decreases the incidence of free radical-induced lipid peroxidation in aged rats. PMID- 12757400 TI - Hydroxyethyl starch (HES) [130/0.4], a new HES specification: pharmacokinetics and safety after multiple infusions of 10% solution in healthy volunteers. AB - OBJECTIVE: To investigate the pharmacokinetics and safety of a daily infusion of 500 mL of hydroxyethyl starch (HES) [130/0.4] 10% solution on 10 consecutive days. STUDY DESIGN AND PARTICIPANTS: An open, one-way, multiple-dose study was performed in 12 healthy male volunteers. Daily infusions over 30 minutes of 500 mL of HES (130/0.4) 10% solution were performed on 10 consecutive days. Plasma and urine HES concentrations were determined repeatedly during the study until 72 hours after the last infusion. RESULTS: Maximum plasma HES concentrations, assessed with geometric means of 7.7 and 7.4 mg/mL, respectively, as well as the time courses of the plasma concentrations were similar on days 1 and 10 of treatment. Plasma HES concentrations 24 hours after the last infusion were 0.48 mg/mL (mean). Total plasma clearance was calculated as 23.7 and 21.8 mL/min on days 1 and 10, respectively. Urinary recoveries of 69% on day 1 and of 70% on day 10 were in good agreement. CONCLUSION: The results clearly demonstrated that there is no relevant accumulation in plasma after repetitive infusion of the medium-molecular weight HES (130/0.4) solution, which exhibits a high renal excretion rate over 10 days. Local as well as systemic tolerability of 10 repeated doses was good. PMID- 12757401 TI - Profile summary. AB - All drugs appearing in the Adis Profile Summary table have been selected based on information contained in R&D Insight trade mark, a proprietary product of Adis International. The information in the profiles is gathered from the world's medical and scientific literature, at international conferences and symposia, and directly from the developing companies themselves. The emphasis of Drugs in R&D is on the clinical potential of new drugs, and selection of agents for inclusion is based on products in late-phase clinical development that have recently had a significant change in status. PMID- 12757402 TI - Abarelix: abarelix-depot-F, abarelix-depot-M, abarelix-L, PPI 149, R 3827. AB - Abarelix [Abarelix-Depot-F, Abarelix-Depot-M, Abarelix-L, PPI 149, R 3827, Plenaxis] is a peptide consisting of natural and artificial amino acids. In females, abarelix is an estrogen production antagonist with potential for the treatment of breast cancer, endometriosis and other reproductive hormone diseases. In males it is a testosterone production antagonist and has potential as hormonal therapy of prostate cancer. Depot formulations of abarelix (abarelix depot-M and abarelix-depot-F) are being developed for hormonally responsive prostate cancer and endometriosis, respectively. Clinical development of the depot formulations is currently being conducted by Praecis Pharmaceuticals, the originators of the agent. A non-depot formulation, abarelix-L, was also being conducted for prostate gland volume reduction. Praecis Pharmaceuticals has entered into a number of licensing agreements covering abarelix. However, all agreements have since been terminated leaving Praecis to develop and commercialise the agent on its own. The terminated agreements include an agreement between Praecis and Roche for the commercialisation of abarelix in the US. This agreement was terminated in November 1998. Praecis Pharmaceuticals also entered into a collaborative agreement with Amgen in March 1999, whereby the companies would develop abarelix and Amgen would commercialise the drug in the US, Canada, Australia, Asia and several secondary markets. However, in September 2001, Praecis and Amgen announced that they were terminating the agreement for all indications. Praecis stated at the time that it remained committed to developing abarelix for both prostate cancer and endometriosis. Amgen had submitted 'Lotestrol' to the US Patent and Trademarks Office as a possible tradename for abarelix-depot-M. Lotestrol may also have been under consideration as a tradename for abarelix-depot-F. Praecis had also sold European, African, Latin American and Middle Eastern rights to abarelix to Sanofi-Synthelabo. However, in October 2001, Sanofi-Synthelabo announced that it had waived its rights to abarelix. Praecis confirmed in December 2000 that it had filed an NDA seeking FDA approval for abarelix in the US. In January 2001, the FDA granted the abarelix application priority review status. However, in June 2001, the FDA rejected the NDA for prostate cancer. The FDA requested that Praecis use existing data from the completed trials to analyse the allergic reactions that occurred in a small subset of patients. The FDA also expressed concerns over the lack of maintenance of testosterone suppression beyond the 3-month timeframe that occurred in a subset of patients. In February 2003, Praecis announced the re submission of its NDA to the US FDA. The submission seeks approval for the use of abarelix in a defined subpopulation of advanced prostate cancer patients for whom the current hormonal therapies are not appropriate. Praecis plans to submit its regulatory application in Europe during the second quarter of 2003. Following the completion of a phase I/II trial of abarelix-L in prostate gland volume reduction, a phase IIIb study of the depot formulation was initiated in September 2001. The trial is comparing the effects of neoadjuvant hormonal therapy with depot formulations of leuprorelin or abarelix for prostate gland volume reduction. Abarelix-L is no longer mentioned on Praecis' website, suggesting that development of this formulation is no longer being pursued. The Financial Times (ft.com) reported in May 2001 that approximately 12 new anti-cancer agents are expected to be approved by the FDA through to the end of 2002, with the potential to generate total sales of US dollars 2.6 billion--abarelix is one of these products. The paper quoted analysts at Salomon Smith Barney predicting that abarelix could reach sales of US dollars 120 million for the indication of prostate cancer. However, in June 2001 the FDA rejected Praecis Pharmaceuticale FDA rejected Praecis Pharmaceuticals' NDA filing; this was later re-submitted in February 2003. A year earlier, in May 2000, the Financial Times (ft.com) stated that Credit Suisse First Boston had forecast abarelix to reach peak sales of 1 billion US dollars . Other analysts, at SG Cowen, predicted annual sales of US dollars 200 million for the first 3 years; however, this could increase to 1 billion US dollars if abarelix is also approved for the indication of endometriosis. Abarelix' main competitors at the time were said to be Lupron [TAP Pharmaceuticals], Viadur [Alza] and Zoladex [AstraZeneca]. PMID- 12757403 TI - Anidulafungin: ECB, LY 303366, V-echinocandin, VEC, VER 002, VER-02. AB - Vicuron Pharmaceuticals (formerly Versicor Inc.) is developing anidulafungin [LY 303366, ECB, V-echinocandin, VEC, VER-02, VER 002], a lipopeptide echinocandin B derivative, for IV treatment of mycoses. Anidulafungin acts against fungal infection by inhibiting beta-1,3-glucan synthase, an enzyme essential for cell wall formation. Anidulafungin was originally developed for oral use by Eli Lilly and was undergoing phase II clinical trials in the UK and the US for the treatment of Candida, Aspergillus and Pneumocystis carinii infections. However, Eli Lilly discontinued development of the oral formulation due to poor oral bioavailability. In May 1999, Versicor obtained exclusive worldwide commercialisation rights to anidulafungin with responsibility for its development and clinical registration. Under the terms of the agreement, Eli Lilly received a signing fee, and will receive milestone payments upon future development of anidulafungin and royalties on future sales. Eli Lilly also retains an option for the development of an oral formulation of the compound. On 3 March 2003, Versicor Inc. of Fremont (California, USA) and Biosearch Italia SpA of Milan (Italy) announced the completion of a merger agreement, whereby Biosearch was merged with and into Versicor in a stock-for-stock exchange valued at US dollars 260.7 million. The combined company temporarily kept the name Versicor until the new name, Vicuron Pharmaceuticals, was announced on 26 March 2003. In January 2003, Versicor announced that positive results from a phase II trial for anidulafungin IV treatment involving 120 patients in the US with invasive candidiasis/candidaemia, have led to another double-blind, randomised phase III trial being conducted in the US, Canada and Europe for this indication. This additional phase III trial will enrol approximately 300 patients to investigate the efficacy of IV anidulafungin (200 mg loading dose followed by 100 mg maintenance dose) versus IV fluconazole for 10 to 42 days. Vicuron Pharmaceuticals also plans to seek approval for invasive candidiasis/candidaemia in Europe and Canada in the second half of 2003. PMID- 12757404 TI - CDP 571: anti-TNF monoclonal antibody, BAY 103356, BAY W 3356, Humicade. AB - CDP 571 [anti-TNF monoclonal antibody, BAY 103356, BAY W 3356, Humicade] is a recombinant humanised antibody directed against tumour necrosis factor (TNF). CDP 571 has an advantage over the mouse/human chimera anti-TNF-alpha antibody, nerelimomab, in that it is suitable for multiple dosing as it is not so immunogenic. Celltech constructed CDP 571 by grafting the section of mouse antibody that recognises TNF onto a human IgG4 antibody. In the third quarter of 1999, Celltech merged with Chiroscience to form Celltech Chiroscience. In January 2000, Medeva was merged into Celltech Chiroscience, which was renamed as Celltech Group. The research division of Celltech Group is now called Celltech R&D (formerly Celltech Chiroscience Discovery) and the manufacturing and marketing division is called Celltech Pharmaceuticals (formerly Celltech Medeva Pharma). Celltech has completed two phase III trials, involving around 670 patients with moderate-to-severe Crohn's disease; however, both these trials failed to meet their primary endpoints. Biogen and Celltech group will review the scope of their collaboration following additional analysis of the phase III data and discussions with regulatory authorities. The Celltech Group intends to devote significant resources towards enhancing the capability of Celltech Pharmaceuticals to market CDP 571 and other new drugs (such as CDP 860 and CDP 870) as specialised hospital products. Phase II trials were underway in the United Kingdom for use of the drug in the treatment of type 2 diabetes mellitus. However, these trials have also been discontinued. Celltech Group is no longer developing CDP 571 for septic shock, based on negative results with the related compound nerelimomab. The compound was in phase III trials for septic shock in France, Belgium, the United Kingdom, Germany and the US. Celltech also plans to investigate the use of CDP 571 in psoriasis via a collaboration with Biogen (USA). In January 2002, AFX (Agence France-Presse and the Financial Times Group) reported that analysts at Morgan Stanley have forecast Humicade trade mark to reach sales of 250 million US dollars in 2008--at that time, the market value for anti-TNF products to treat rheumatoid arthritis and Crohn's disease will exceed 4 billion US dollars, according to Morgan Stanley. PMID- 12757405 TI - Decitabine: 2'-deoxy-5-azacytidine, Aza dC, DAC, dezocitidine, NSC 127716. AB - Decitabine [NSC 127716, DAC, dezocitidine, Aza dC, 2'-deoxy-5-azacytidine] is a deoxycytidine and cytarabine derivative with potent antileukaemic activity, which was originated by Pharmachemie. This antimetabolite is able to induce in vitro gene activation and cellular differentiation by a mechanism involving DNA hypomethylation. SuperGen acquired worldwide rights to decitabine from Pharmachemie in the third quarter of 1999 for 4 million US dollars worth of SuperGen shares and income from manufacture upon the launch of decitabine. SuperGen announced in May 2000 that it had entered a Cooperative Research and Development Agreement (CRADA) with the US National Cancer Institute (NCI). SuperGen will supply decitabine to the NCI, which will initiate and sponsor clinical trials in patients with solid tumours and haematological malignancies. The NCI will also conduct studies on decitabine's mechanism of action. In 2002, the US FDA has granted decitabine orphan drug status for the treatment of myelodysplastic syndromes and sickle cell anaemia. In February 2003, the European Commission granted orphan drug status to decitabine for myelodysplastic syndrome. Decitabine has also received orphan drug status in the US as a host-protective agent in the treatment of AML. Decitabine has been studied in solid tumours as well as in different types of leukaemia. In several phase II studies it has been shown to have very limited efficacy against solid tumours. However, decitabine has shown better activity in the treatment of haematological malignancies such as acute myeloid leukaemia (AML), chronic myeloid leukaemia (CML) and myelodysplastic syndrome (preleukaemia). In March 2001, SuperGen announced that it had begun patient enrolment into its pivotal open-label phase III trial of decitabine in advanced myelodysplastic syndrome patients. The study, which will compare decitabine with standard care therapy, will be conducted at 15 medical centres in the US and will enrol a total of 160 patients. In March 2003, SuperGen announced that patient enrolment was complete. The study, which will compare decitabine with standard care therapy, will be conducted at 22 medical centres in the US and will enrol a total of 160 patients. A European pivotal trial is also underway for the same indication, and is aiming to enrol 220 patients. A phase I/II trial of 8 patients, designed to establish safety and efficacy in the treatment of sickle cell anaemia, has been completed at the University of Illinois, USA. Plans for additional studies of decitabine as a treatment for sickle cell anaemia are underway. Decitabine is also undergoing phase II clinical trials in Canada, for the treatment of non-small cell lung cancer, and in the US for chronic myeloid leukaemia and prostate cancer. Glasgow University in Scotland has conducted preclinical trials in chemotherapy-resistant ovarian and colon cancers. The results suggest that decitabine administration may reverse chemotherapy resistance in these cancers. SuperGen was issued a US patent (No. 6 191 119) in 2001 covering the use of decitabine in combination with rubitecan and antibiotic agents, including doxorubicin. PMID- 12757406 TI - Dexanabinol: dexanabinone, HU 211, PA 50211, PRS 211007, sinnabidol. AB - Dexanabinol [HU 211, dexanabinone, sinnabidol, PA 50211, PRS 211007] is a synthetic, non-psychotropic tetrahydro-cannabinoid. Dexanabinol lacks cannabimimetic activity, and is a functional antagonist of the NMDA receptor with antioxidant and anti-tumour necrosis factor-alpha properties. Dexanabinol is in clinical trials for traumatic brain injury (head injuries), glaucoma and mild cognitive impairment, and is being investigated preclinically for its potential in the treatment of multiple sclerosis. Pharmos has a licensing agreement with the Hebrew University, Israel, and is seeking a partner for development and commercialisation of the dexanabinol family of compounds. The Financial Times (ft.com) reported in March 2001 that the market for brain trauma could be worth approximately 500 million US dollars, according to estimates by Pharmos. The company was said to have 26 million US dollars in capital at the time, most of which would be used taking dexanabinol through regulatory submission, according to the Financial Times. PMID- 12757407 TI - EGb 761: ginkgo biloba extract, Ginkor. AB - EGb 761 [Ginkgo biloba extract EGb 761, Rokan, Tanakan, Tebonin] is a standardised extract of Ginkgo biloba leaves and has antioxidant properties as a free radical scavenger. A standardised extract of Ginkgo biloba leaves is a well defined product and contains approximately 24% flavone glycosides (primarily quercetin, kaempferol and isorhamnetin) and 6% terpene lactones (2.8-3.4% ginkgolides A, B and C, and 2.6-3.2% bilobalide). Ginkgolide B and bilobalide account for about 0.8% and 3% of the total extract, respectively. Other constituents include proanthocyanadins, glucose, rhamnose, organic acids, D glucaric and ginkgolic acids. EGb 761 promotes vasodilation and improves blood flow through arteries, veins and capillaries. It inhibits platelet aggregation and prolongs bleeding time. EGb 761, which was originated by Dr Willmar Schwabe Pharmaceuticals (Dr Willmar Schwabe Group), has been available in Europe as a herbal extract since the early 1990s. However, products containing EGb 761 are not approved for use by the US FDA. As a dietary supplement, Nature's Way in the US distributes and markets a standardised extract of Ginkgo biloba leaves (the EGb 761 Formula) under the name Gingold Nature's Way. The French company Beaufour Ipsen and its German subsidiary Ipsen Pharma are co-developing EGb 761 with Dr Willmar Schwabe Group. Beaufour-Ipsen (France) is developing EGb 761 as Tanakan, Dr Willmar Schwabe Pharmaceuticals (Germany) as Tebonin and Ipsen Pharma (Germany) as Rokan. Intersan was formerly developing EGb 761 in Germany, but Intersan appears to have been merged into Ipsen Pharma. However, there has been no recent development for these indications. In the UK and other European countries, the cardioprotective effects of EGb 761 in myocardial ischaemia and reperfusion are being investigated in preclinical studies. The psychological and physiological benefits of ginkgo are said to be based on its primary action of regulating neurotransmitters and exerting neuroprotective effects in the brain, protecting against or retarding nerve cell degeneration. Ginkgo also benefits vascular microcirculation by improving blood flow in small vessels and has antioxidant activity. There has been conflicting evidence about the benefits of ginkgo, e.g. the ginkgo clinical trial published in August 2002 in JAMA concluded that a leading ginkgo supplement did not produce measurable benefits for memory in healthy adults over 60, although a month earlier, another study concluded that the same ginkgo extract is effective in helping normal healthy older adults in memory and concentration. However, in December 2002, the Cochrane Collaboration, the world's most respected scientific reviewer of clinical trials in medicine, concluded that the published literature strongly supports the safety and potential benefits of ginkgo in treating memory loss and cognitive disorders associated with age- related dementia. A phase II study of EGb 761 in combination with fluorouracil is in progress in Germany in patients with pancreatic cancer. German researchers are investigating the potential of EGb 761 for the treatment of sudden deafness and tinnitus in clinical studies. EGb 761 was undergoing preclinical development for the potential treatment of diabetes in France, diabetic neuropathies in Russia, and cancer in Brazil. However, there has been no recent development for these indications. Beaufour-Ipsen has expressed the intention to license out its diabetes projects that may include EGb 761. PMID- 12757408 TI - Hyaluronidase (Vitrase)--ISTA: hyaluronidase--ISTA pharmaceuticals. AB - ISTA Pharmaceuticals (formerly Advanced Corneal Systems) has developed an ophthalmic injectable formulation of highly purified hyaluronidase [ovine hyaluronidase, Vitrase] for the initial treatment of vitreous haemorrhage and diabetic retinopathy. Hyaluronidase is a naturally occurring enzyme that digests certain forms of carbohydrate molecules called proteoglycans. The current medical treatment for vitreous haemorrhage is vitrectomy, an invasive surgical procedure that may result in future cataract formation, retinal detachment or other complications. There are currently no approved drug therapies for vitreous haemorrhage. ISTA believes that an injection of Vitrase causes the vitreous to liquefy, thereby promoting the clearance of vision-distorting blood. The elimination of blood helps to restore vision and provides an ophthalmologist with an unobstructed view of the retina, allowing the doctor to diagnose and treat the underlying cause of the hemorrhage. In mid-1997, Advanced Corneal Systems (now ISTA Pharmaceuticals) formed a Singapore subsidiary called Visionex to develop and market the company's technologies in Southeast Asia and China. In March 2000, ISTA completed the acquisition of Visionex. Also in March 2000, subsidiaries of Allergan obtained marketing, sales and distribution agreements from ISTA for Vitrase worldwide, except Mexico (until April 2004) and Japan. ISTA will split Vitrase profits equally with Allergan and receive royalties on sales in non-US countries. ISTA is responsible for all costs of product development, preclinical studies and clinical trials, of Vitrase and may receive up to 35 million US dollars in milestone payments from Allergan upon the achievement of specified regulatory and development objectives. In December 2001, Otsuka gained exclusive rights to develop, market and commercialise Vitrase in Japan. In July 2002, ISTA announced that it has entered into an agreement with Cardinal Health for the manufacture of commercial quantities of Vitrase. The agreement covers the US, Canada, Japan and the European Union. Cardinal Health will also provide manufacturing-related information for the US New Drug Application (NDA). Sophia Laboratories distribute Vitrase in Mexico. The US FDA designated Vitrase as a fast track product in October 1998, which means the FDA will facilitate the development and expedite the review of the product. Vitrase has being investigated in two multinational, randomised, placebo-controlled, phase III trials in patients with severe vitreous haemorrhage. One was conducted in the US, Mexico and Canada (North American trial) with an enrolment of 750 patients. The second trial was conducted in Europe, Brazil, Australia and South Africa and enrolled 556 patients. In March 2002, ISTA began unmasking the data, revealing that although preliminary efficacy results did not show any statistically significant improvement in the primary endpoint, clinically relevant improvements in visual acuity and a decrease in the density of vitreous haemorrhage were observed in patients treated with a 55IU dose of Vitrase, compared with placebo treated patients. In December 2002, the FDA accepted the NDA for Vitrase for filing. The FDA's Dermatologic and Ophthalmic Drugs Advisory Committee reviewed the Vitrase NDA on 17 March 2003 and voted 8 to 4 that there was insufficient statistical evidence to support the use of Vitrasefor the treatment of vitreous haemorrhage. However, the Committee did recognise that in certain patient subgroups, the benefits of Vitrase therapy outweighed the potential risks. The FDA has recommended that ISTA provide additional analyses from the two pivotal phase III trials conducted. In April 2003, the FDA issued an approvable letter for Vitrase for the treatment of vitreous haemorrhage. ISTA anticipates that the FDA will complete its review of the Vitrase NDA anete its review of the Vitrase NDA and issue the results during the second half of 2003. In addition, ISTA plans to submit a marketing approval application with the European Medical Evaluation Agency (EMEA) in the first half of 2003. A phase II trial in Singapore was being conducted by Visionex. However, in March 2000, ISTA completed the acquisition of Visionex. In its Securities and Exchange Commission (SEC) filing, as at 31 December 2002, ISTA stated that the continued development of Vitrase for diabetic retinopathy will be dependent upon a number of factors including the FDA's evaluation of Vitrase for the treatment of vitreous hemorrhage, the successful completion of any additional clinical trials for the diabetic retinopathy, and the continuing assessment of the market opportunity for this indication compared with other product opportunities that ISTA may be pursuing at the time. ISTA is also developing hyaluronidase products for the treatment of cataracts (Keratase) and keratoconus (Keraform). PMID- 12757409 TI - Marimastat: BB 2516, TA 2516. AB - Marimastat [BB 2516, TA 2516] is a second-generation anticancer drug originally developed with British Biotech in Europe and North America. It is an orally active metalloprotease inhibitor of the same class as batimastat, and is the first compound in this class to have completed a pivotal clinical trial. Marimastat also has collagenase- and angiogenesis-inhibiting properties. British Biotech and Schering-Plough have signed an agreement enabling the latter to develop and market marimastat in North America and Europe. Under the terms of the agreement, British Biotech will receive an up-front license fee of 4 million US dollars and a 4 million US dollars equity investment in British Biotech by Schering-Plough. Schering-Plough holds rights to marimastat in all countries other than the Far East and Japan. The two companies are considering asking the FDA for accelerated approval in gastric cancer based on the secondary endpoint of progression-free survival. Marimastat is licensed to Tanabe Seiyaku in Japan, where phase II clinical trials are underway for the treatment of advanced gastric cancer and lung cancer. Further phase II trials in other tumour types are planned. The commencement of phase II trials in Japan resulted in a milestone payment of 5 million US dollars to British Biotech from Tanabe Seiyaku. Tanabe Seiyaku also holds rights to marimastat in the Far East. Marimastat has been in pivotal phase III trials in glioblastoma, breast, ovarian and small and non-small cell lung cancer, but these trials have all been discontinued because marimastat failed to show superior efficacy over either standard chemotherapy or placebo. Results from the marimastat 131 trial in patients with glioblastoma, for example, indicated that marimastat was no better than placebo at prolonging survival in these cancer patients. In June 2000, when the results of this study were released, shares in British Biotech fell 21.6% to just 19 pence per share. The phase III trial in small cell lung cancer was discontinued when the results of study 140 were released in February 2001 showing that marimastat was not significantly more effective than placebo in prolonging the survival of small cell lung cancer patients. The results of this study were consistent with those reported in study 117. British Biotech has also conducted a phase III placebo controlled study of marimastat as monotherapy in patients with inoperable gastric cancer at 37 centres throughout Europe. Results from this trial indicated that it did not achieve its primary endpoint of a statistically significant survival benefit over placebo. However, data collected during the follow-up period have shown increases in survival benefit in the treatment group in addition to a significant improvement in disease-free progression, the secondary endpoint of the trial. Development of marimastat for this indication is ongoing. In May 2001, British Biotech reported data from an interim analysis of results from the remaining phase III study in pancreatic cancer (study 183) that showed no patient benefit for marimastat recipients compared with gemcitabine. However, these results did not meet stopping criteria and the study continues under the guidance of Schering-Plough. The multicentre trials are being conducted in the US, Canada and the European Union. The phase III trial of marimastat in combination with carboplatin that was being conducted in patients with ovarian cancer was discontinued because British Biotech realised that the design of the trial was insufficient for registration in the US or Europe. Altogether, seven phase III studies have failed to meet their primary end-points, but the company has stated that the effectiveness of marimastat is more likely to be seen in patients with less advanced disease. Phase II trials in prostate and head and neck cancer are still underway in the US. PMID- 12757410 TI - Oxymorphone--Endo/Penwest: EN 3202, EN 3203. AB - Penwest Pharmaceuticals and Endo Pharmaceuticals are jointly developing an oral, controlled-release opioid analgesic oxymorphone [EN 3202] using Penwest's TIMERx proprietary drug delivery technology. The product is being developed for twice-a day dosing in patients with moderate to severe pain. TIMERx is a controlled release technology based on an agglomerated hydrophilic matrix, which consists of the polysaccharides locust bean gum and xanthan gum. The technology provides a full spectrum of controlled-release profiles lasting >/=4 h. In February 2003, the US FDA has announced that it accepted the NDA for oxymorphone extended release tablets, oxymorphone ER, for the treatment of moderate to severe pain in patients requiring continuous opioid therapy for an extended period of time. Late in 2002, the Oxymorphone formulation successfully completed a phase III clinical trial in patients with osteoarthritis pain. Another randomised, double-blind, placebo-controlled trial was completed in 127 patients with moderate to severe pain resulting from surgery in the knee. Results demonstrated that patients receiving EN 3202 treatment had significantly superior pain relief compared with placebo recipients. An immediate-release formulation of oxymorphone (EN 3203) has also been accepted by the US FDA. PMID- 12757411 TI - Hepatocyte growth factor activates endothelial nitric oxide synthase by Ca(2+)- and phosphoinositide 3-kinase/Akt-dependent phosphorylation in aortic endothelial cells. AB - Hepatocyte growth factor (HGF) causes endothelium-dependent vasodilation, but its relation to endothelial nitric oxide synthase (eNOS) activity remains to be elucidated. Treatment of bovine aortic endothelial cells with HGF increased eNOS activity within minutes, accompanied by an increase of activity-related site specific phosphorylation of eNOS. The phosphorylation was completely abolished by pretreatment of the cells with a phosphoinositide 3-kinase (PI3K) inhibitor (wortmannin) and by transfection of dominant-negative Akt, and the enzyme activity was inhibited by wortmannin. In addition, eNOS activity and phosphorylation were abolished by pretreatment of the cells with an intracellular Ca(2+)-chelator, bis-(o-aminophenoxy)ethane-N,N,N',N'-tetra-acetic acid tetrakis(acetoxymethyl ester) (BAPTA/AM), with a suppression of Akt phosphorylation. These results suggest that HGF stimulates eNOS activity by a PI3K/Akt-dependent phosphorylation in a Ca(2+)-sensitive manner in vascular endothelial cells. PMID- 12757412 TI - An antitumour lectin from the edible mushroom Agrocybe aegerita. AB - An antitumour lectin (named AAL) consisting of two identical subunits of 15.8 kDa was isolated from the fruiting bodies of the edible mushroom Agrocybe aegerita using a procedure which involved precipitating the extract by addition of (NH(4))(2)SO(4), ion exchange chromatography on DEAE-Sepharose Fast Flow, gel filtration chromatography on Sephacryl S-200 HR and finally purification on a GF 250 HPLC column. Amino acid analysis of the N-terminus and an internal fragment indicated that the sequences of the two fragments were QGVNIYNI and Q(K)PDGPWLVEK(Q)R respectively. AAL showed strong inhibition of the growth of human tumour cell lines HeLa, SW480, SGC-7901, MGC80-3, BGC-823, HL-60 and mouse sarcoma S-180. AAL also inhibited the viability of S-180 tumour cells in vivo. Analysis by Hoechst 33258 staining, MitoSensor Kit and flow cytometry showed that AAL induced apoptosis in HeLa cells. TUNEL (terminal transferase deoxytidyl uridine end labelling) analysis of slides of tumour tissues excised from BALB/c mice also demonstrated the apoptosis-induction activity of the lectin. Furthermore, AAL was shown to possess DNase activity in assays using plasmid pCDNA3 and salmon sperm DNA. Based on the results obtained in these assays, we conclude that AAL exerts its antitumour effects via apoptosis-inducing and DNase activities. PMID- 12757430 TI - 'and even evidence-based librarianship'? PMID- 12757429 TI - Chylomicron-remnant-like particles inhibit the basal nitric oxide pathway in porcine coronary artery and aortic endothelial cells. AB - The effects of chylomicron remnants on the activity of basally produced nitric oxide (NO) from porcine coronary artery rings and porcine aortic endothelial cells were studied by investigating the effects of chylomicron-remnant-like particles (CMR-LPs) containing porcine apolipoprotein E on the vessel tone of porcine coronary arteries and on cGMP release by aortic endothelial cells. CMR LPs were oxidized by incubation with CuSO(4) (10 microM) for 18 h at 37 degrees C. N (omega)-nitro-L-arginine (L-NOARG) and oxidized CMR-LPs (oxCMR-LPs), but not native CMR-LPs, increased the vessel tone of static porcine coronary artery rings (increase in tone as a percentage of the tone induced by depolarizing Krebs Henseleit solution: L-NOARG, 14.24 +/- 2.09; oxCMR-LPs, 4.98 +/- 0.88; and native CMR-LPs, 0.47 +/- 0.21). L-NOARG, endothelium removal and oxCMR-LPs also all significantly increased the maximum relaxation of the vessels to S -nitroso- N acetyl-DL-penicillamine. In addition, oxCMR-LPs reduced the amounts of cGMP released by porcine aortic endothelial cells into the culture medium from 116 +/- 12.0 to 84.2 +/- 11.6 fmol/microg of cellular protein, mimicking the effects of L NOARG. These results indicate that oxCMR-LPs, but not native CMR-LPs, inhibit the activity, production or release of NO from unstimulated porcine coronary and aortic endothelial cells. oxCMR-LPs mimicked the addition of L-NOARG and endothelium removal in these experimental systems, suggesting that the lipoproteins were interfering with the L-arginine/NO pathway. This study provides further evidence to support a role of chylomicron remnants in the development of atherosclerosis. PMID- 12757431 TI - Information skills training: a systematic review of the literature. AB - The objectives of this study were to undertake a systematic review to determine the effectiveness of information skills training, to identify effective methods of training and to determine whether information skills training affects patient care. A systematic review, using an iterative approach to searching, was employed. Studies selected for inclusion in the review were critically appraised using a tool used in previous reviews. A tabular approach was used to provide a summary of each paper allowing synthesis of results. One thousand, three hundred and fifty-seven potentially relevant papers were located. On the basis of titles and abstracts, 41 potentially relevant studies were identified for potential inclusion. Further reading and application of the inclusion criteria left 24 studies for critical appraisal and inclusion in the review. Study designs included randomised controlled trials, cohort designs and qualitative studies. The majority of studies took place in US medical schools. Wide variations were found in course content and training methods. Eight studies used objective methods to test skills, two compared training methods and two examined the effects on patient care. There was limited evidence to show that training improves skills, insufficient evidence to determine the most effective methods of training and limited evidence to show that training improves patient care. Further research is needed in a number of areas. PMID- 12757432 TI - Clinical librarianship: a systematic review of the literature. AB - Clinical librarianship (CL), currently receiving renewed interest world-wide, seeks to provide quality-filtered information to health professionals at the point of need to support clinical decision-making. This review builds upon the work of Cimpl (Bulletin of the Medical Library Association 1985, 73, 21-8) and attempts to establish the evidence base for CL. The objectives were to determine, from the literature, whether CL services are used by clinicians, have an effect on patient care, and/or clinicians' use of literature in practice and/or are cost effective. The methodology used was a systematic review of the literature, following, where possible, the NHS Centre for Reviews and Dissemination (CRD) framework. Modifications to this methodology included the resources searched, and the critical appraisal checklist (CriSTAL) used. Two hundred and eighty-four unique references were retrieved. Seventeen (16 unique) evaluative and a further 33 descriptive studies met the inclusion criteria. The quality of reporting of the literature was generally poor. CL programmes appear to be well-used and received by clinicians. However, there is insufficient evidence available on their effect on patient care, clinicians' use of literature in practice, and their cost-effectiveness, thus highlighting the need for further high-quality research. PMID- 12757433 TI - Educational workshop improved information-seeking skills, knowledge, attitudes and the search outcome of hospital clinicians: a randomised controlled trial. AB - A double-blind randomised controlled trial was conducted on a group of Hong Kong hospital clinicians. The objective was to test if a three-hour educational workshop (with supervised hands-on practice) is more effective (than no training) to improve clinical question formulation, information-seeking skills, knowledge, attitudes, and search outcomes. The design was a post-test-only control group; recruitment by stratified randomization (by profession), blocked at 800. End-user training was more effective than no training in improving clinical question formulation, in raising awareness, knowledge, confidence and use of databases, but had made no impact on preference for secondary databases. It changed the attitude of clinicians to become more positive towards the use of electronic information services (EIS). Participants had higher search performance and outcomes (satisfaction with information obtained (NNT = 3), EIS satisfaction (NNT = 3) and success in problem solving (NNT = 4)). The workshop improved knowledge and skills in evidence-based searching, but this effect gradually eroded with time. Search logs confirmed that follow-up is required if effects are to be sustained. Longer effects on search behaviours appear to be positive. A randomised controlled trial is valuable in identifying cause-and-effect relations and to quantify the magnitude of the effects for management decision-making. PMID- 12757434 TI - The randomised controlled trial design: unrecognized opportunities for health sciences librarianship. AB - OBJECTIVE: to describe the essential components of the Randomised Controlled Trial (RCT) and its major variations; to describe less conventional applications of the RCT design found in the health sciences literature with potential relevance to health sciences librarianship; to discuss the limited number of RCTs within health sciences librarianship. METHODS: narrative review supported to a limited extent with PubMed and Library Literature database searches consistent with specific search parameters. In addition, more systematic methods, including handsearching of specific journals, to identify health sciences librarianship RCTs. RESULTS: While many RCTs within the health sciences follow more conventional patterns, some RCTs assume certain unique features. Selected examples illustrate the adaptations of this experimental design to answering questions of possible relevance to health sciences librarians. The author offers several strategies for controlling bias in library and informatics applications of the RCT and acknowledges the potential of the electronic era in providing many opportunities to utilize the blinding aspects of RCTs. RCTs within health sciences librarianship inhabit a limited number of subject domains such as education. This limited scope offers both advantages and disadvantages for making Evidence-Based Librarianship (EBL) a reality. CONCLUSIONS: The RCT design offers the potential to answer far more EBL questions than have been addressed by the design to date. Librarians need only extend their horizons through use of the versatile RCT design into new subject domains to facilitate making EBL a reality. PMID- 12757435 TI - Clear-cut?: facilitating health librarians to use information research in practice. AB - In 1999, staff at the universities of Sheffield and Oxford commenced an unfunded project to examine whether it is feasible to apply critical appraisal to daily library practice. This aimed to establish whether barriers experienced when appraising medical literature (such as lack of clinical knowledge, poor knowledge of research methodology and little familiarity with statistical terms) might be reduced when appraising research within a librarian's own discipline. Innovative workshops were devised to equip health librarians with skills in interpreting and applying research. Critical Skills Training in Appraisal for Librarians (CRISTAL) used purpose-specific checklists based on the Users' Guides to the Medical Literature. Delivery was via half-day workshops, based on a format used by the Critical Appraisal Skills Programme. Two pilot workshops in Sheffield and Oxford were evaluated using a brief post-workshop form. Participants recorded objectives in attending, their general understanding of research, and whether they had read the paper before the workshop. They were asked about the length, content and presentation of the workshop, the general format, organization and learning environment, whether it had been a good use of their time and whether they had enjoyed it. Findings must be interpreted with caution. The workshops were enjoyable and a good use of time. Although the scenario selected required no clinical knowledge, barriers remain regarding statistics and research methodology. Future workshops for librarians should include sessions on research design and statistics. Further developments will take forward these findings. PMID- 12757436 TI - A Delphi study to determine nursing research priorities in the North Glasgow University Hospitals NHS Trust and the corresponding evidence base. AB - The aims of the study are to identify and prioritize research questions of importance to nurses in North Glasgow University Hospitals NHS Trust and to investigate and describe the existing evidence base for the highest priority questions identified. The consensus method known as the Delphi technique was used. Systematic searching of the literature for each question identified from the research priorities enabled the existing evidence base to be characterized. The following priorities were identified as the most important (in descending order of importance): Recruitment and Retention of Nursing Staff; Staff Stress & Morale; Training & Education of Nursing Staff; Infection Control; and Pain Management. The Delphi technique proved a useful method to determine and prioritize research areas in nursing. PMID- 12757437 TI - LISTENing to healthcare students: the impact of new library facilities on the quality of services. AB - Following a low assessment of 'Learning resources' provision by the Quality Assurance Agency, the librarian of Homerton College, School of Health Studies commenced the LISTEN Project, a long-term study to monitor the effects of planned interventions on the quality of library provision. Surveys of entry-to-register student nurses & midwives were conducted in 1999 and 2001 by extensive questionnaires, inviting Likert-scaled and free text responses. Following a college relocation, students made greater than expected use of a new health studies library in Cambridge, and significantly less use of the local teaching hospital library. Using both a satisfaction index and a non-parametric test of mean scores, student evaluation of library services in Cambridge significantly improved following relocation. The physical accommodation and location of library services remain important to healthcare students. Identifiable improvements to the quality of services, however, will overcome initial resistance to change. Education providers must ensure the best mix of physical and electronic services for students who spend much of their time on clinical placement. PMID- 12757438 TI - Towards a Cochrane Information Retrieval Methods Group: a progress report. PMID- 12757439 TI - Journal clubs for continued professional development. PMID- 12757440 TI - Facilitating evidence-based librarianship: a Canadian experience. PMID- 12757441 TI - Facilitating evidence-based librarianship: a UK experience. PMID- 12757444 TI - Microbial products in allergy prevention and therapy. PMID- 12757445 TI - Epidermal growth factor receptor - but not histamine receptor - is upregulated in seasonal allergic rhinitis. AB - BACKGROUND: We were interested in exploring the molecular mechanisms underlying the observed difference in histamine (H) responsiveness between seasonal allergic rhinitic (SAR) and nonrhinitic (NR) subjects. We hypothesized that SAR subjects express higher nasal mucosal histamine receptor 1 (H1R) and 2 (H2R) levels than do NR subjects. In addition, we examined expression of genes involved in regulating the glandular response, including epidermal growth factor (EGF), EGF receptor (EGFR), and mucins (Muc5Ac and Muc5B). METHODS: Fourteen subjects, seven SAR and seven NR, were provoked during pollen season with doubling doses of H (0.125-8.0 mg/ml). Nasal airway resistance (NAR) was measured by active posterior rhinomanometry. Provocation was halted when NAR exceeded 150% of baseline. Prior to provocation, nasal scrapings were obtained and mRNA quantified using two-step real-time PCR. RESULTS: The mean PD50 (concentration of H producing a 50% increase in NAR) was significantly lower in the SAR than NR group (0.36 vs 1.32 mg/ml; P < 0.05). The ratio of relative gene copy numbers between the SAR and NR groups were as follows: H1R, 0.85 (P = 0.52); H2R, 0.67 (P = 0.35); EGF, 1.02 (P = 0.93), and EGFR, 103.5 (P < 0.05). CONCLUSIONS: There were no significant differences in H1R or H2R mRNA levels between SAR and NR subjects in-season, despite observed differences in H reactivity. SAR subjects, however, did show a significant elevation in EGFR expression, consistent with the observation of mucus hypersecretion in allergic rhinitis. PMID- 12757446 TI - Nasal blockage and urinary leukotriene E4 concentration in patients with seasonal allergic rhinitis. AB - BACKGROUND: Cysteinyl-leukotrienes have been reported to have a primary role in the induction of nasal blockage of allergic rhinitis. However, there has been little experimental evidence that substantiates the relationship between nasal blockage severity and urinary leukotriene E4 (U-LTE4) concentration in patients with seasonal allergic rhinitis (SAR). METHODS: The concentrations of urinary mediators in 20 SAR patients were measured using an enzyme immunoassay to determine the relationship between nasal blockage severity and U-LTE4 concentration in patients with SAR. RESULTS: The basal U-LTE4 concentration was significantly higher in SAR patients with severe nasal blockage than in those with mild nasal blockage and in healthy control subjects. Although U-LTE4 concentrationwas significantly higher in patients with both asthma and SAR than in SAR patients with mild nasal blockage, no significant difference in the U-LTE4 concentration between patients with both asthma and SAR and SAR patients with severe nasal blockage was found. There was a significant correlation between U LTE4 and urinary 9alpha11beta-prostoglandin F2 (9alpha11betaPGF2) concentrations (rs = 0.51, P = 0.02) in SAR patients. CONCLUSIONS: Although specific sites and cells of cysteinyl-leukotriene biosynthesis could not be determined in this study, severe nasal blockage is associated with the increased excretion level of U-LTE4. PMID- 12757447 TI - Comparison of the effects of desloratadine 5-mg daily and placebo on nasal airflow and seasonal allergic rhinitis symptoms induced by grass pollen exposure. AB - BACKGROUND: Nasal congestion is a chronic symptom of seasonal allergic rhinitis (SAR) that is often difficult to treat with antihistamines. Desloratadine, a new, potent, H1-receptor antagonist has been shown to decrease nasal congestion in clinical trials and to maintain nasal airflow in response to grass pollen exposure. We compared the effects of desloratadine 5 mg and placebo on nasal airflow, nasal secretion weights and SAR symptoms, including nasal congestion, in patients exposed to grass pollen in an environmental exposure unit. METHODS: Forty-six grass pollen allergic SAR patients received desloratadine or placebo for 7 days, followed by a 10-day washout, and then crossed over to the other treatment for 7 days. A 6-h allergen exposure was performed at the end of each treatment period. RESULTS: Desloratadine was significantly superior to placebo in maintaining nasal airflow (P or= 32 micromol/l). The median value of induced cough counts after a HCA challenge was 11 coughs in patients with ICS and was a significantly enhanced cough response compared to that of the patients with NCS and healthy controls (four coughs, respectively). CONCLUSIONS: A simple cough provocation test using a HCA challenge may be useful for detecting ICS. It also suggests that hyperreactive cough reflexes may be one of the mechanisms of inducing chronic cough. PMID- 12757449 TI - Herbal supplements and skin testing: the lack of effect of commonly used herbal supplements on histamine skin prick testing. AB - BACKGROUND: The use of herbal supplements is common, yet little is known about their pharmacologic properties. The purpose of this study was to assess the effects of 23 commonly used herbal supplements on histamine skin prick testing (SPT). METHODS: Fifteen healthy volunteers participated in a double-blind, placebo-controlled, single-dose, crossover study. Wheal and flare responses to SPT with histamine phosphate (1 mg/ml) were measured before and 4 h after administration of each of the 23 popular herbal supplements, fexofenadine (60 mg) and placebo. Wheal and flare areas were recorded with tracings performed 10 min after the prick test and measured with a PC-digitizer using stereometric software. RESULTS: Fexofenadine significantly suppressed the wheal (P < 0.001) and flare (P = 0.02) areas compared with placebo. None of the herbal supplements caused significant suppression of the wheal and flare areas compared with placebo (P > 0.10). CONCLUSION: When taken in single-doses, the popular herbal supplements tested did not significantly affect the histamine skin response. Therefore, it seems unnecessary for clinicians to ask patients to discontinue these herbal supplements prior to allergy skin testing. PMID- 12757450 TI - Combined skin prick and patch testing enhances identification of peanut-allergic patients with atopic dermatitis. AB - BACKGROUND: Food atopy patch tests (APTs) are considered a useful tool for the diagnosis of food allergy. Hypersensitivity to peanuts has not been investigated by means of APTs so far. METHODS: APTs and skin prick tests (SPTs) with peanuts were performed in 136 atopic dermatitis (AD) patients. Relevance of positive and negative responses to these tests was assessed by repeated open challenges with peanuts. RESULTS: Nine percent of our AD patients reacted to the challenge. Positive responses to APTs were recorded in 19% of the patients, whereas in 12% positive SPTs were observed. APTs were more frequently positive in subjects with eczematous responses after challenge with respect to those with urticarial reactions. SPT reactivity proved to be higher in patients above 12 years of age, whereas APT positivity was more frequent in children under 6 years. APT sensitivity proved significantly higher than SPT sensitivity, in particular in children under 12 years of age. On the contrary, SPT specificity and positive predictive value were significantly higher with respect to those of APT in the age group of subjects under 6 years of age. CONCLUSIONS: Our data suggest that APTs with peanuts may represent a useful integration to standard testing modalities employed for the diagnosis of peanut allergy in AD patients. PMID- 12757451 TI - Distribution of house dust mite allergen: comparing house dust mite allergen levels in dust samples collected from different sites on living room floors with smooth coverings. AB - BACKGROUND: The distribution of house dust mite allergen (Der p1) in living rooms with smooth floor coverings, as measured in the middle compared with the border of the floor was investigated. It was hypothesized that activity causes displacement of Der p1, from the middle towards the border. METHODS: Dust samples from the middle and border of 50 floors with smooth coverings were collected and analysed on Der p1 content in a standardized way. RESULTS: The Der p1 exposure expressed as per unit area (ng/m2) showed that border samples contained significantly more Der p1 compared with middle samples (median: 2.57 vs 0.27, respectively, P = 0.023). Presence of pets and presence of more than two inhabitants increased the difference. When expressed as per unit weight of dust (ng/g), significant differences were only detected when comparing Der p1 content of samples collected in households with three or more inhabitants [median: 2 (border) vs 53 (middle), respectively; P = 0.035]. CONCLUSIONS: The Der p1 is unequally distributed on living room floors with smooth coverings, most likely because of displacement of dust from the middle towards the border due to activity. Expression as ng/g of dust and ng/m2 could not obviously be interchangeable. PMID- 12757453 TI - Lettuce anaphylaxis: identification of a lipid transfer protein as the major allergen. AB - BACKGROUND: Allergy to plant-derived foods is associated with birch pollinosis in central and northern Europe. Symptoms elicited are usually limited to the oropharyngeal system. By contrast, in the Mediterranean area, allergy to the same foods manifests more frequently with systemic reactions caused by nonspecific lipid transfer proteins (nsLTP), independently of an associated pollinosis. OBJECTIVE: We sought to investigate the pattern of immunoglobulin E (IgE) binding protein bands implicated in lettuce allergy, in particular the presence of an nsLTP. METHODS: Consecutive lettuce allergic patients were selected. Determination of serum-specific IgE, immunoblot, and inhibition experiments were performed in order to study the pattern of IgE binding proteins and the potential cross-reactivity to pollens. Inhibition studies with recombinant allergens were conducted to identify the lettuce allergens. The major allergen was subjected to N-terminal amino acid sequencing. RESULTS: Fourteen patients were diagnosed as being allergic to lettuce. All were sensitized to Platanus pollen. Ten of them showed specific IgE to a lettuce protein of 9-kDa. The IgE binding to this protein was completely inhibited by the cherry-LTP and peach extract. The N terminal sequence of the 9-kDa protein showed a high degree of amino acid sequence identity to other nsLTPs. A clear partial cross-reactivity was observed between lettuce-LTP and Platanus-pollen extract. CONCLUSIONS: An LTP has been demonstrated to be a major allergen in patients suffering from lettuce allergy. PMID- 12757452 TI - Cross-reactivity between the major Parietaria allergen and rotavirus VP4 protein. AB - BACKGROUND: The present study investigates immunological cross-reactivity between Par o 1, the major pollen allergen of Parietaria, and the VP4 protein of rotavirus, a microorganism that is world-wide the main etiological agent of gastroenteritis in children. METHODS: IgG and IgE cross-reactivity was assessed by direct binding and competitive inhibition assays (ELISA and DARIA), using recombinant VP4 from rhesus infectious rotavirus (RR), synthetic peptides and Par o 1-specific antibodies affinity purified from pooled and individual human sera. RESULTS: Antibodies specifically binding Par o 1, affinity purified from the sera of 35 individuals with skin test positivity to Parietaria and from 14 pools, were extensively cross-reactive with RRVP4. Cross-reactive binding was specifically inhibited by synthetic peptides derived from the C-terminal sequences of the VP4 proteins from human and rhesus infectious rotavirus. CONCLUSIONS: This study reports the first evidence of cross-reactivity between an allergen and a viral antigen. PMID- 12757454 TI - Accumulation of CCR4-expressing CD4+ T cells and high concentration of its ligands (TARC and MDC) in bronchoalveolar lavage fluid of patients with eosinophilic pneumonia. AB - BACKGROUND: Th2 cells are thought to be involved in eosinophilic inflammation of the lung. CC chemokine receptor 4 (CCR4) has been identified as a specific receptor for both thymus and activation-regulated chemokine (TARC) and macrophage derived chemokine (MDC), and is preferentially expressed on Th2 cells. OBJECTIVE: Our aim was to evaluate the role of Th2 cells in the lung of patients with eosinophilic pneumonia (EP). METHODS: The concentrations of TARC, MDC, and interleukin (IL)-5 were measured in bronchoalveolar lavage fluid (BALF) by ELISA. Proportion of CCR4-expressing CD4+ T cells (CCR4+ CD4+ T cells) was determined by flow cytometry. RESULTS: TARC and MDC concentrations in BALF were higher in patients with EP than in normal subjects. The proportion of CCR4-expressing cells among CD4+ T cells was higher in BALF than in peripheral blood of patients with EP. There was a significant correlation between the number of CCR4+ CD4+ T cells and the levels of TARC, MDC, and IL-5 in BALF of patients with EP. CONCLUSIONS: Our data suggest that Th2 cells, which express CCR4 and its ligands (TARC and MDC), contribute to the pathogenesis of EP in the lung. PMID- 12757455 TI - A dual long-term effect of breastfeeding on atopy in relation to heredity in children at 4 years of age. AB - BACKGROUND: The long-term effect of early feeding on atopic sensitization is still unsolved. The aim of this study was to evaluate the long-term effect of breastfeeding on atopy in groups of 4-year-old children stratified by atopic heredity. METHODS: We collected four groups of 4-year-old children from a birth cohort: two groups with differing backgrounds of atopic heredity, all exclusively breast-fed for at least 3 months; and two groups with differing atopic heredity, but all fed with cow's milk-based formula during their first weeks. The data were collected with a questionnaire, skin prick testing, and measurement of serum total and allergen-specific IgE levels. RESULTS: Breastfeeding significantly decreased the risk of allergic rhino-conjunctivitis [odds ratio (OR) 0.41, 95% confidence interval (CI) 0.18-0.95] and sensitization to furred pets, as measured by skin prick results, in children with atopic heredity, whereas in children without atopic heredity, breastfeeding was related to an increased risk of symptomatic atopy (OR 2.57, 95% CI 1.16-5.70), and high serum IgE values. A significant interaction was found between heredity and breastfeeding. CONCLUSIONS: The long-term effect of breastfeeding was dual: in children with atopic heredity, breastfeeding protected against atopy, whereas in children without atopic heredity, it increased the risk of atopy. PMID- 12757456 TI - Removal of house-dust mites and Der p I during laundering of bedding. PMID- 12757458 TI - Allergy to Ailanthus altissima (tree of heaven) pollen. PMID- 12757457 TI - Low rate of cockroach sensitivity in Italian pre-school children. PMID- 12757459 TI - Egg allergy--to be or not to be boiled. PMID- 12757460 TI - Corn-induced hypersensitivity pneumonitis. PMID- 12757461 TI - Food allergy to alcohol. PMID- 12757462 TI - Prolonged cyclosporin-A treatment for severe chronic urticaria. PMID- 12757463 TI - Systemic allergic reaction by a human insulin analog. PMID- 12757464 TI - Type-I hypersensitivity to ceftriaxone and cross-reactivity with cefalexin and ampicillin. PMID- 12757465 TI - CD63 expression by flow cytometry in the in vitro diagnosis of allergy to omeprazole. PMID- 12757466 TI - Delayed-type hypersensitivity rash from ibuprofen. PMID- 12757467 TI - Rapid oral desensitization to isoniazid, rifampin, and ethambutol. PMID- 12757468 TI - Contrast-enhanced magnetic resonance angiography: development and optimization of techniques for paramagnetic and hyperpolarized contrast media. AB - Contrast-enhanced magnetic resonance angiography (CE-MRA) is a diagnostic method for imaging of vascular structures based on nuclear magnetic resonance. Vascular enhancement is achieved by injection of a contrast medium (CM). Studies were performed using two different types of CM: conventional paramagnetic CM, and a new type of CM based on hyperpolarized (HP) nuclei. The effects of varying CM concentration with time during image acquisition were studied by means of computer simulations using two different models. It was shown that a rapid concentration variation during encoding of the central parts of k-space could result in signal loss and severe image artifacts. The results were confirmed qualitatively with phantom experiments. A postprocessing method was developed to address problems with simultaneous enhancement of arteries and veins in CE-MRA of the lower extremities. The method was based on the difference in flow-induced phase in the two vessel types. Evaluation of the method was performed with flow phantom measurements and with CE-MRA in two volunteers using standard pulse sequences. The flow-induced phase in the vessels of interest was sufficient to distinguish arteries from veins in the superior-inferior direction. Using this method, the venous enhancement could be extinguished. The possibility of using HP nuclei as CM for CE-MRA was evaluated. Signal expressions for a flow of HP CM imaged with a gradient echo sequence were derived. These signal expressions were confirmed in phantom experiments using HP 129Xe dissolved in ethanol. Studies were also performed with a new CM based on HP 13C. The CM had very long relaxation times (T1, in vivo/T2, in vivo approximately 38/1.3 s). The long relaxation times were utilized in imaging with a fully balanced steady-state free precession pulse sequence (trueFISP), where the optimal flip angle was found to be 180 degrees. CE-MRA with the 13C-based CM in rats resulted in images with high vascular SNR (approximately 500). CE-MRA is a useful clinical tool for diagnosing vascular disease. With the development of new contrast media, based on hyperpolarized nuclei for example, there is a potential for further improvement in the signal levels that can be achieved, enabling a standard of imaging of vessels that is not possible today. PMID- 12757469 TI - Fas expression on T cells and sFas in relapsing-remitting multiple sclerosis. AB - OBJECTIVES: To investigate the proportions of peripheral blood CD4+/Fas+ and CD8+/Fas+ cells and serum sFas levels in relapsing-remitting multiple sclerosis (RRMS) patients with relapses (active RRMS), those without relapses (stable RRMS), and controls over 1 year. MATERIAL AND METHODS: Sixteen RRMS patients and 10 controls were tested monthly. Cells were analyzed by dual immunofluorescence and the sFas levels by ELISA. There were 14 relapses which occurred 1223 days after the last control visits. The measurements performed at these visits in the active RRMS patients were considered as relapse-related, while the rest were regarded as relapse-unrelated. RESULTS: In active RRMS patients the median of CD4+ Fas+ to total CD4+ and CD8+ Fas+ to total CD8+ from relapse-related measurements were higher than the median from relapse-unrelated measurements (P=0.003, 0.004, respectively). The median of CD4+ Fas+ to total CD4+ from relapse-unrelated measurements in active RRMS was higher compared with stable RRMS (P = 0.005) and controls (P = 0.004). The sFas level from relapse-unrelated measurements was also higher in active RRMS than in stable RRMS (P = 0.04) and in controls (P = 0.004). CONCLUSIONS: We suggest that increased expression of Fas antigen on CD4+ subset and increased serum sFas level are valuable markers of clinical activity in MS. PMID- 12757470 TI - Effects of psychological group therapy in patients with multiple sclerosis. AB - OBJECTIVES: The aim of this pilot study was to evaluate a psychological therapy program used in the treatment of multiple sclerosis (MS) and including cognitive/behavioral strategies, relaxation training and physical exercise. MATERIAL AND METHODS: The participants were 29 patients with MS recruited from an outpatient unit; 14 patients were assigned to the 7-week psychological therapy group (one session per week), the remainder formed a control group. Before and immediately after the course of therapy and after a 2-month follow-up, the participants completed a series of questionnaires measuring factors such as depression, anxiety, coping and body image. RESULTS: Compared with the control group the therapy group showed long-term improvements in depressive stress coping style and a short-term improvement in "vitality and body dynamics". CONCLUSION: Further studies should investigate the differential effects of specific units of the therapy program and how the short-term improvements in "vitality and body dynamics" could be maintained for longer periods. PMID- 12757471 TI - The milder phenotype of the dystrophin gene double deletions. AB - OBJECTIVES: This study aimed to examine the genotypephenotype correlation in Duchenne muscular dystrophy (MD) patients with double deletion (Ddel) mutations in comparison with those having single deletions (Sdel). MATERIALS AND METHODS: The study included 250 Duchenne/Becker MD male patients from whom the 10 Ddel patients were compared with 20 Sdel subjects of same age and disease durations. The patients were subjected to neurological examination including functional disability grading scale (FDGS), molecular analysis of the dystrophin gene and immunohistochemical studies of some muscle biopsies. RESULTS: The mean FDGS value in the Ddel group was lower than that in Sdel patients. The Ddel patients had partial expression of dystrophin in their skeletal muscles, while Sdel cases showed complete absence of the protein. CONCLUSION: Patients with double deletion mutations within the dystrophin gene have a milder phenotype than patients harboring single deletions at either major or minor hot spots of the gene. PMID- 12757472 TI - Changes of cerebral blood flow velocities during enhanced external counterpulsation. AB - OBJECTIVES: Intra-aortic counterpulsation is the most frequently used cardiac assist device. However, there are only few studies of the effects of counterpulsation on cerebral blood flow and these report conflicting outcomes. The new enhanced external counterpulsation (EECP) technique reproduces non invasively the effects of intra-aortic counterpulsation. In this study, we evaluated effects of EECP on blood pressure (BP) and on cerebral flow velocity (CBFV). SUBJECTS AND METHODS: Twenty-three healthy controls and 15 atherosclerotic patients each underwent a 5-min session of EECP. Before, during and after EECP we monitored heart rate, beat-to-beat radial artery BP and CBFV. RESULTS: EECP induced a second increase in BP and CBFV during diastole with a significant increase of mean BP and a decrease of systolic BP in patients and controls. Mean CBFV increased in both groups during the first 5 s of EECP. After 3 min of EECP, diastolic CBFV was still higher than at baseline, but systolic CBVF was lower than at baseline; mean CBFV was as low as before EECP in the patients and lower than the baseline values in the controls. Three minutes after ending EECP, mean and systolic BP were lower in the patients than the corresponding baseline values. Otherwise, CBFV and BP values did not differ from baseline in patients and controls. CONCLUSION: Cerebral autoregulation ensures the constancy of cerebral blood flow even though EECP creates marked systemic changes. In the patients, the decrease of BP after EECP with maintained CBFV indicates an improved BPCBFV relation and a more economic autoregulation. PMID- 12757473 TI - Endogenous protectant kynurenic acid in amyotrophic lateral sclerosis. AB - OBJECTIVES: Excitotoxicity may play a role in neurodegeneration in amyotrophic lateral sclerosis (ALS). Kynurenic acid (KYNA), an endogenous antagonist of excitatory amino acid receptors, may inhibit excitotoxic lesions. The aim of this study was to determine the concentration of KYNA in ALS patients. MATERIAL AND METHODS: KYNA was measured by high-performance liquid chromatography in the serum and cerebrospinal fluid (CSF) from ALS and control patients. RESULTS: Our study revealed that CSF KYNA concentration was significantly higher in patients with bulbar onset of ALS compared to controls, and compared to patients with limb onset of the disease. CSF KYNA was also higher in patients with severe clinical status compared to controls. Serum KYNA was significantly lower in ALS patients with severe clinical status compared to controls, and compared to patients with mild clinical status. There were no significant differences in CSF and serum KYNA concentration between the whole ALS group of patients and controls. There was no difference in CSF KYNA concentration between males and females, and there was no correlation between KYNA concentration and age of patients, and duration of ALS. CONCLUSIONS: An increased CSF KYNA concentration in patients with bulbar onset of ALS and in patients with severe clinical status may indicate neuroprotective role of KYNA against excitotoxicity. The difference of KYNA concentration in CSF of patients with bulbar and limb onset of ALS suggests that these two variants of motor neuron disease may have different etiopathogenetic mechanisms. PMID- 12757474 TI - Genetics of familial amyloidotic polyneuropathy in a Hong Kong Chinese kindred. AB - Familial amyloidotic polyneuropathy type 1 (FAP1, MIM176300) is an autosomal dominant disease caused by mutations in the transthyretin (TTR) gene. An extended Chinese kindred of FAP1 was first reported in Hong Kong in 1989, three of the four histologically proven subjects have deceased. TTR gene mutations were not studied then. A DNA-based diagnosis was performed on FAP1 by restriction analysis and direct DNA sequencing was carried out on a symptomatic member of this family who had undergone a liver transplantation. It showed a substitution of thymine by cytosine in the second base of codon 30 in exon 2 of the TTR gene, with the creation of a novel HhaI restriction endonuclease site. Valine is substituted by alanine (V30A) in the mutant TTR. Both restriction analysis and direct sequencing revealed the same mutation in one of the two asymptomatic siblings. This mutation was first reported in a FAP1 family of German descent. PMID- 12757475 TI - Ischemic infarct involving all arterial territories of the thalamus. AB - Ischemic infarcts of the thalamus involve one or two of its four arterial territories that are usually supplied by the posterior cerebral (PCA) and the posterior communicating (PCoA) arteries. We report a patient who suffered ischemic infarcts in all arterial territories of the right thalamus. Magnetic resonance (MR) angiography showed an occlusion of the right PCA and failed to visualize a PCoA. We assume that the absence of a relevant thalamic blood supply deriving from the PCoA enabled PCA occlusion to cause infarcts in all thalamic territories. PMID- 12757476 TI - MRI in ALS patients. PMID- 12757498 TI - Near-patient testing of haemostasis in the operating theatre: an approach to appropriate use of blood in surgery. AB - Several haemostasis point-of-care (POC) monitors are now available in the operating theatre. Two of these are widely used; the Coaguchek and the thromboelastograph (TEG), but they have been developed in very different ways. Bedside-activated partial thromboplastin time and prothrombin time performed with the Coaguchek monitor seem to be reliable and have been used to build algorithms for transfusion decision making. They have been developed in close collaboration with haemostasis groups and therefore gain a benefit from these links. Conversely, TEG provides very important information except it has never been validated. The number of collaborative studies with biologists has to increase in order to implement the use of TEG in the routine practice. PMID- 12757499 TI - In vivo and in vitro comparison of platelets stored in either synthetic media or plasma. AB - Since 1980, several synthetic media have been developed for the storage of platelets for transfusion. At present, platelets suspended in approximately 70% synthetic medium and 30% plasma can be stored for at least 5 days at very stable pH levels, generally pH 6.8-7.2. Present knowledge suggests that synthetic media should contain at least acetate, citrate, phosphate, potassium and magnesium. Future studies will probably result in the inclusion of other components to this list. Glucose for platelet metabolism will generally be supplied by carryover of plasma from the original platelet preparation. In addition, improved plastic containers for the storage of platelets will probably facilitate the introduction of new synthetic media. In six studies comparing synthetic media with plasma as the storage environment, and involving patients with intensive chemotherapy for haematological malignancies, the clinical outcome in terms of corrected count increments (CCI) generally indicated similar results. Three studies suggested significant reduction of the incidence of transfusion reactions of platelets suspended in synthetic media as compared with plasma. For future comparisons of platelet storage in either plasma or new synthetic media, additional platelet survival and recovery studies, as well as patient-transfusion studies, will be needed as in vitro data may not always reflect the clinical outcome. This will add further knowledge to data from the present few clinical studies available that compare storage of platelets in either synthetic media or plasma. PMID- 12757500 TI - Evaluation of the de-selection of men who have had sex with men from blood donation in England. AB - BACKGROUND AND OBJECTIVES: The Blood Services of the UK permanently de-select men who have had sex with men (MSM) from donating blood. The rationale for this has been questioned. This article attempts to evaluate whether this selection criterion does contribute to blood safety. MATERIALS AND METHODS: Data about transfusion-transmissible infections, in particular about human immunodeficiency virus (HIV) infection, were used to evaluate whether de-selection of MSM meets the aims of donor selection. Models were constructed to estimate the risk of HIV infectious donations entering the blood supply should this criterion be changed. RESULTS: Many assumptions were required to generate estimates of the risk of HIV infection entering the blood supply. The accuracy of the estimates is therefore uncertain and the probable ranges around the estimates were wide. However, by using the most probable assumptions, our models suggested that de-selection of MSM for 12 months since the last sexual contact, or complete removal of this selection criterion, would be expected to increase the risk of HIV-infectious donations entering the blood supply in England by approximately 60% (from the current risk of 0.45 per year to 0.75 per year) and 500% (to 2.5 per year), respectively. The increase in numbers of non-infected donations would be relatively small--less than 2% of donations. The probability of a relatively high frequency of other sexually transmissible blood-borne infections also currently favours maintaining permanent de-selection of MSM, irrespective of the risk of HIV-infectious donations. Current compliance with this selection criteria was estimated to be 95%. CONCLUSIONS: Based on current knowledge, accepting blood donations from MSM would probably increase the risk of transfusion-transmission of HIV and of other blood-borne infections. Good compliance with this criterion has contributed greatly to the safety of blood transfusions in England. Better communication about donor selection, to maintain and improve compliance with this and other selection criteria, is recommended. Other risk groups are gaining in relative importance for the risk of transfusion-transmitted HIV infection, and ongoing evaluation of all donor-selection criteria is also recommended. PMID- 12757501 TI - Estimation of the risk of hepatitis B virus, hepatitis C virus and human immunodeficiency virus infectious donations entering the blood supply in England, 1993-2001. AB - BACKGROUND AND OBJECTIVES: The frequency of hepatitis B virus (HBV), hepatitis C virus (HCV) or human immunodeficiency virus (HIV) infectious donations entering the blood supply in England is too low to monitor using observational studies. The expected frequency of infectious donations can be estimated and these estimates may be used to contribute to monitoring of blood safety and used in the design of strategies to decrease the risk of transfusion-transmitted infections. MATERIALS AND METHODS: The prevalence and incidence of hepatitis B surface antigen (HBsAg), and antibodies to HCV and HIV (anti-HCV and anti-HIV, respectively) in donors in England, between 1993 and 2001, were used together with data about the length of negative 'window-periods' of current assays for each of these markers and data about test performance, to estimate the number of infectious donations that enter the blood supply. The risks were calculated separately for donations from new donors and from repeat donors, and for the three time periods 1993-95, 1996-98 and 1999-01. RESULTS: The estimated frequency of infectious donations entering the blood supply in England, between 1993 and 2001 was 1 in 260,000 for HBV and 1 in 8 million for HIV. For HCV, the frequency of infectious donations was 1 in 520,000 during 1993-98 and fell to 1 in 30 million during 1999-2001 when all donations were tested for HCV RNA. The frequency of HBV- and HCV-infectious donations entering the blood supply fell over these 9 years: the frequency of HIV-infectious donations remained essentially unchanged. The risk from donations from new donors was found to be approximately sevenfold higher than the risk from donations from repeat donors. CONCLUSIONS: The risks of HBV-, HCV- or HIV-infectious donations entering the blood supply in England are very low, and have decreased since 1993. Although the accuracy of these estimates is imperfect, mainly owing to uncertainty in some assumptions and to small numbers of infections, they provide some quantification of the risk of HBV, HCV or HIV transmission by transfusion, and allow comparison of the magnitude of these risks for each infection and over time. The methods we have used have been developed and improved from previously published methods. PMID- 12757502 TI - Superiority of minipool nucleic acid amplification technology for hepatitis B virus over chemiluminescence immunoassay for hepatitis B surface antigen screening. AB - BACKGROUND AND OBJECTIVES: The Japanese Red Cross (JRC) have developed a fully automated multiplex (MPX) nucleic acid amplification technology (NAT) system for hepatitis B virus (HBV), hepatitis C virus (HCV) and human immunodeficiency virus 1 (HIV-1). This is used to test serologically negative blood units from volunteer, non-remunerated donors. The system utilizes a 50-sample pool for NAT screening with an input volume of each pool. This results in a significantly higher sensitivity for hepatitis B than that seen with highly sensitive hepatitis B surface antigen (HBsAg) testing. MATERIALS AND METHODS: From 1 February 2000 to 15 October 2001, over 11 million donations, which were serologically negative, were tested using the MPX NAT system. Donations found to be HBV DNA positive were further tested by using the chemiluminescence immunoassay (CLIA). RESULTS: Out of 181 HBV DNA-positive donations, 96 (53%) and 76 (42%) were negative by individual enzyme immunoassay (EIA) and CLIA testing, respectively. CONCLUSIONS: The sensitivity of the 50-sample pool MPX NAT system was higher than that of individual HBsAg screening by CLIA. By adopting this NAT-screening system, the JRC has improved the safety of the blood supply and maintained supply across Japan. PMID- 12757503 TI - High frequency of hepatitis B virus infection in patients with beta-thalassemia receiving multiple transfusions. AB - BACKGROUND AND OBJECTIVES: Hepatitis B virus (HBV) may occasionally be transmitted through transfusion of blood units that are hepatitis B surface antigen (HBsAg) negative but HBV DNA positive. Children with beta-thalassemia are particularly susceptible to HBV because they receive multiple blood transfusions. These children have high infection rates despite vaccination against HBV. Post vaccination infections may be a result of viruses harbouring surface (S)-gene mutations (e.g. G587A) in a region critical for reactivity to antibody to hepatitis B surface antigen (anti-HBs). The true prevalence of HBV in individuals with beta-thalassemia has not been studied previously. PATIENTS AND METHODS: Seventy patients with beta-thalassemia (median age 6 years; range 8 months to 22 years; 49 male), who had received seven to 623 (median 61) units of blood each and three doses (10/20 micro g) of HBV vaccine (Engerix B) before presentation to us, were included in the study; 50 of the 70 patients had received transfusions prior to vaccination. Enzyme-linked immunoassay for serological markers [HBsAg, antibody to hepatitis B core antigen (anti-HBc) and quantitative anti-HBs] and polymerase chain reaction (PCR) followed by Southern hybridization for molecular detection of hepatitis B, was performed on all samples. The PCR-amplified product was cloned, sequenced and the nucleotide and deduced amino acid sequences for the HBV S and polymerase (P) genes were analysed for mutations. RESULTS: Four of 70 (5.7%) individuals with beta-thalassemia were HBsAg positive and 14 (20%) were anti-HBc positive. The prevalence of serological markers increased with number of transfusions (P < 0.01). Of 70 patients, 53 (75.7%) had an anti-HBs titre of > 10 IU/l following vaccination and 17 (24.3%) were non-responders (< 10 IU/l); 22 (31.4%) of the 70 were DNA positive. The frequency of HBV infection in beta thalassemia was similar in vaccine responders and non-responders. The virus was of subtype ayw (genotype D) in the five DNA-positive samples in which a 388 nucleotide region of the S gene was sequenced. Mutations occurred at 13 positions in the S gene and at 10 positions in the P gene. Hydrophobicity plots revealed differences in amino acid regions 117-165 and 195-211. Some of these amino acid substitutions coincided with the putative cytotoxic T-lymphocyte epitopes of both S and P proteins. CONCLUSIONS: A high frequency of HBV infection was seen using molecular methods in thalassemic patients. The frequency of infection was similar in vaccine responders and non-responders. A number of mutations were observed in the S gene, which could have implications for viral replication as well as virus host cell interaction. PMID- 12757504 TI - Prevalence of human erythrovirus B19 DNA in healthy Belgian blood donors and correlation with specific antibodies against structural and non-structural viral proteins. AB - BACKGROUND AND OBJECTIVES: Human parvovirus (erythrovirus) B19 is recognized as a major contaminant of blood and blood products. To reduce the risk of contamination, plasma-pool screening and exclusion of highly viraemic donations are recommended. The objectives of this study were to estimate the prevalence of B19 DNA in our blood-donor population, to determine the appropriate pool size to be tested (taking into account parameters such as prevalence, viral load, test sensitivity, and the efficacy of inactivation procedures), and to correlate viral loads with the serological status of donors as regards antibodies against different viral proteins. MATERIALS AND METHODS: Pools of different sizes were tested for B19, using a sensitive nested polymerase chain reaction (PCR) as well as an simple, un-nested, less sensitive PCR. Positive pools were resolved to the level of individual donations, and the viral load and serological markers were determined. RESULTS: Of 16,859 donations, 27 (one of 625) were found to be B19 DNA positive, with viral loads ranging from 10(2) to > 10(7) IU/ml. Twenty-five of the positive donations were tested for VP-specific anti-B19 antibodies, and eight (32%) were negative for both immunoglobulin (Ig)M and IgG. They were probably collected in the preseroconversion window period or from chronic carriers without detectable antibodies. We regarded the seven (28%) IgM-positive donors as being in the early phase of infection. The remaining 10 (40%) IgM negative, IgG-positive donors were probably carriers of persistent infection (i.e. PCR positive despite the presence of IgG antibodies), as suggested by their low viral loads (< 10(4) IU/ml). Fifteen out of 36 major pools contained one or more contaminated donations. Among these, 12 tested positive by nested PCR and only three by un-nested PCR, this reflecting a viral load of > 10(4) IU/ml. CONCLUSIONS: By testing all donations as pools of 480 by un-nested PCR, and resolving positive pools to identify the responsible donations, it is possible to ensure that the viral load in fractionation pools (5000 donations) remains < 10(3) IU/ml, compatible with the efficacy of inactivation procedures and complying with Food and Drug Administration (FDA) recommendations. PMID- 12757505 TI - Quality assessment of seven types of fresh-frozen plasma leucoreduced by specific plasma filtration. AB - BACKGROUND AND OBJECTIVES: A study was undertaken to determine plasma quality after specific filtration. MATERIALS AND METHODS: Seven types of plasma were tested, after filtration of plasma from filtered or non-filtered whole blood. Leucocyte counting was carried out after a 30-fold concentration of the sample. Twenty-nine parameters (including coagulation testing, proteins, coagulation factors and activation markers) were measured before and after filtration, and after 6 months of storage. RESULTS: After specific plasma filtration, the average residual leucocyte counts were less than 2250/l. In spite of small statistically significant changes in proteins, coagulation factors and complement activation, this study showed that plasma filtration did not alter plasma quality. After 6 months of storage at -30 degrees C, factor VIII recovery varied between 91 and 109%. Haemostasis parameters and activation markers remained within the normal range. CONCLUSIONS: Specific plasma filtration reduced the leucocyte number to < 104 leucocytes/l. The quality of plasma was not altered by the additional step of specific plasma filtration. PMID- 12757506 TI - Will it ever be possible to balance the risk of intracranial haemorrhage in fetal or neonatal alloimmune thrombocytopenia against the risk of treatment strategies to prevent it? AB - BACKGROUND AND OBJECTIVES: Intracranial haemorrhage (ICH) of the fetus or newborn is a severe complication of fetal or neonatal alloimmune thrombocytopenia (FNAIT). In order to attain management decisions to prevent ICH, the risk of ICH in successive pregnancies with thrombocytopenia, with or without a history of ICH, must be established. MATERIALS AND METHODS: We performed a search of medline for ICH cases in untreated FNAIT pregnancies. After exclusion of cases with confounding factors, 24 reports, describing 62 pregnancies of 27 mothers, were eligible. In addition, two mothers with five pregnancies were included from our own case records. Observational studies were examined to estimate the risk of ICH in subsequent FNAIT pregnancies without a history of ICH. Finally, medline was searched for complication rates in the treatment of FNAIT pregnancies. RESULTS: In 52% of the ICH cases, a previous sibling suffered from ICH. The recurrence rate of ICH in the subsequent offspring of women with a history of FNAIT with ICH was 72%[confidence interval (CI): 46-98%] without inclusion of fetal deaths and 79% (CI: 61-97%) with inclusion of fetal deaths. In 48% of the ICH cases, the previous sibling had thrombocytopenia but not ICH. Population studies revealed an overall ICH risk in thrombocytopenic infants of 11% (CI: 0.8-23%) without inclusion of fetal deaths and 15% (CI: 1.5-19%) with inclusion of fetal deaths. Assuming occurrence in 48%, the risk of ICH in a subsequent pregnancy following a history of FNAIT without ICH, was estimated to be 7% (CI: 0.5-13%). Invasive treatment strategies carry a risk of 2.8% (CI: 1.2-4.4%) on complications. CONCLUSIONS: The number of eligible publications on ICH in untreated FNAIT pregnancies is strikingly limited. The recurrence rate is high. As sufficient data on successive FNAIT cases without ICH are lacking, the occurrence of ICH in pregnancies with thrombocytopenia, but without ICH in a previous sibling, cannot be predicted. We estimate this risk to be 7%. This risk must be balanced against the risk of interventions in treatment strategies. PMID- 12757508 TI - Regarding 'Meta-analysis of randomized trials comparing the risk of postoperative infection between recipients of allogeneic and autologous blood transfusion' by Vamvakas et al. PMID- 12757507 TI - A novel DI*A allele without the Band 3-Memphis mutation in Amazonian Indians. AB - BACKGROUND AND OBJECTIVES: The blood-group antigens Dia and Dib are carried on erythrocyte band 3 and are defined by a single amino acid substitution at position 854 (Leu for Dia and Pro for Dib). The Band 3-Memphis variant has a point mutation (166A>G) in the SLC4A1 gene, which encodes the amino acid substitution Lys56Glu. Two types of Band 3-Memphis, variants I and II, are distinguished by their susceptibility to covalent labelling with 4,4' diisothiocyanato-1,2-diphenylethane-2,2'-disulphonic acid (H2DIDS). Memphis II is more readily labelled than Memphis I or normal band 3. It is reported that Memphis II is associated with Dia. In a study designed to determine the frequency of the DI*A/DI*B and 166A>G polymorphisms in different populations in Brazil, we found a new DI*A allele. MATERIALS AND METHODS: We studied DNA samples from 70 Amazonian Indians, 71 individuals of Japanese descent, 93 random Brazilian blood donors and 84 blacks with sickle cell disease. Polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) analyses were performed on all samples, using MspI for DI*A/DI*B (exon 19) and MnlI for 166A>G (exon 4). Exon 4 and exon 19 from four outliers were sequenced. RESULTS: Among Amazonian Indians, DI*A and 166G mutations both had a high frequency (0.57 and 0.54, respectively). In individuals of Japanese descent, these alleles were moderately frequent (0.07 and 0.19, respectively). We identified a new allele with DI*A and 166A (56Lys) in four Amazonian Indians. CONCLUSIONS: Our results revealed that DI*A does not have a strict association with 166G. They also show the relevance of testing a cohort of different populations. PMID- 12757512 TI - Unusual phenotype of cis-AB. PMID- 12757510 TI - Solvent-detergent-treated plasma may be cost-effective. PMID- 12757513 TI - Fetomaternal alloimmune thrombocytopenia. PMID- 12757516 TI - Do HLA antibodies cause hemolytic transfusion reactions or decreased RBC survival? PMID- 12757514 TI - Transfusion medicine illustrated. Lack of effect of 2 M urea on Jk(a-b-) cells. PMID- 12757517 TI - How much do we know about the platelet transfusion threshold? PMID- 12757519 TI - Frequency of HBV DNA detection in US blood donors testing positive for the presence of anti-HBc: implications for transfusion transmission and donor screening. AB - BACKGROUND: An estimate of the rate of HBV DNA-positive, anti-HBc-positive units is important for evaluating the need for anti-HBc donor screening, especially in the context of HBV NAT. STUDY DESIGN AND METHODS: HBsAg EIA-nonreactive, anti-HBc reactive (Corzyme, Abbott Laboratories) specimens were retrieved from a repository and were retested for anti-HBc (with PRISM HBcore, Abbott Laboratories, currently under FDA review) and anti-HBs (with PRISM Ausab, Abbott Laboratories, research assay). HBV DNA testing using a PCR assay with a greater than 95 percent detection rate of less than 50 copies per mL was performed on a subset of specimens that were PRISM HBcore-reactive and were anti-HBs- negative or reactive at less than 100 IU per L. RESULTS: A total of 395 of 1231 specimens eligible by our serologic criteria were tested by PCR. Four anti-HBs-negative specimens were PCR-positive with estimated HBV DNA copy numbers of 10 per 30 copies per mL in two specimens and 50 to 100 copies per mL in two others. The HBV DNA detection rate in anti-HBs-negative specimens was 3.7 percent, and the projected rate among all Corzyme-reactive specimens was 0.24 percent, leading to an estimated yield of 1 HBV DNA-positive, anti-HBc-positive unit in 49,000 units that were otherwise eligible for transfusion (95% CI, 1 in 16,600-1 in 152,600). CONCLUSIONS: Anti-HBc screening detects HBsAg EIA-negative, HBV-infected donors at a rate comparable to the estimated residual risk for HBV window-period infections. The low viral load in the HBV DNA-positive samples suggests that minipool NAT will not detect most potentially infectious units from anti-HBc positive donors. PMID- 12757520 TI - Prevalence of transfusion-transmissible viral infections in first-time US blood donors by donation site. AB - BACKGROUND: Understanding the donor base, infectious disease prevalence, and donation loss at various blood donation sites will help maximize blood collection efforts and blood availability. STUDY DESIGN AND METHODS: Using donation data collected at five US blood centers, the prevalence of HIV, HTLV, HBsAg, and HCV in first-time whole-blood donations at 10 donation sites was evaluated: military, education, religious, professional, industry, services, community, health care, government, and fixed sites. Donation loss from screening test reactivity at each donation site was also evaluated. RESULTS: During the study, 1.2 million first time whole-blood donations were collected. Military and education sites had a low prevalence of all viral markers, except for HBsAg, which was highest at education sites. Variations in viral marker prevalence among donation sites were partially explained by donor demographic differences. Donation loss varied by donation site, ranging from 3.3 percent at education sites to 6.4 percent at industry sites, indicating differential efficiency of blood collection efforts. CONCLUSION: Different rates of positive viral test results and donation loss in first-time whole-blood donors were observed at various types of donation sites. This information may be useful in estimating the yield of usable units from specific blood drives and in allocating resources to meet blood center collection goals. PMID- 12757521 TI - HCV screening in blood donations using RT-PCR in mini-pool: the experience in Spain after routine use for 2 years. AB - BACKGROUND: The aim of this study is to evaluate the feasibility of implementing a commercial HCV RNA RT-PCR screening method and provide data on the prevalence and incidence rates of hepatitis C in Spain. STUDY DESIGN AND METHODS: Five transfusion centers participated in the study, covering 34.1 percent of the country's total number of donations. All the centers evaluated the sensitivity and characteristics of a commercial RT-PCR reagent kit designed for pool testing (Cobas AmpliScreen HCV v2.0), for which serial dilutions of HCV WHO International Standard 96/790 and preseroconversion samples were used. The data obtained from this technique, employed routinely from May 1999 to June 2001 in 22- to 48-unit mini-pools, are presented in this study. RESULTS: An overall 95-percent detection limit was obtained either at 69 IU per mL when 0.2 mL volume of plasma was extracted (used to analyze individual units), or at 20 IU per mL, when viral particles were pelleted from 1 mL plasma (as used for screening in mini-pool) Three HCV-RNA-positive anti-HCV-negative donations were identified out of 1,015,482 screened donations. One of these had an initially undisclosed risk of HCV sexual transmission and carried a low viral load of 104.2 IU per mL HCV RNA. The analysis of first-time (FT) donations during the period of study (21.3% of the total) indicated an average prevalence rate of 2.05 per 103 FT donors (of which 1.55/103 FT donors were RNA positive); the residual risk calculated on repeat (RPT) donors was 3.91 per 106 donations (serology) or 0.59 per 106 donations (serology + NAT), and the predicted NAT yield estimate was 4.2 per 106 FT + RPT donations. CONCLUSIONS: The commercial RT-PCR reagent kit complies with the current European and FDA recommendations on sensitivity and can be easily implemented on a routine basis. The results obtained by the five transfusion centers on the predicted NAT yield (1/302,000 RPT donations or 1/237,000 FT + RPT donations) are very close to the published estimates corresponding to a larger area of our country (1/237,000 RPT donations) and are somewhat higher than, though in line with, the observed NAT yield (1/338,000 FT + RPT donations). PMID- 12757522 TI - The cost-effectiveness of NAT for HIV, HCV, and HBV in whole-blood donations. AB - BACKGROUND: The risk of viral infection associated with blood transfusion is lower than ever before because of aggressive screening and testing practices. NAT technology has lowered that risk even further but at an additional cost to the health-care system. STUDY DESIGN AND METHODS: Marginal cost-effectiveness of NAT for HIV, HCV, and HBV in whole-blood donations was calculated with a previously published Markov decision model. This model was updated with disease incidence data from all 2001 American Red Cross whole-blood donations as well as window period data from the Retrovirus Epidemiology Donor Study (REDS). RESULTS: Whole blood donation NAT for HIV and HCV is expected to cost between 155 US dollars million (minipool NAT) and 428 million US dollars(single-donation NAT) per year in the US and avert 4 to 7 HIV infections and 56 to 59 HCV infections. Adding HBV NAT would be expected to avert 9 to 37 HBV infections at an additional cost of between 39 million US dollars and 130 million US dollars per year. Overall, NAT would cost between 4.7 million US dollars and 11.2 million US dollars per quality adjusted life-year saved. Discontinuing HIV p24 antigen and HBc testing would offset this somewhat. CONCLUSIONS: The cost-effectiveness of whole-blood NAT is poor. The testing cost would need to decrease significantly to bring the cost effectiveness in line with most other accepted medical practices. PMID- 12757523 TI - Temporal trends in the prevalence of HIV and other transfusion-transmissible infections among blood donors in northern Thailand, 1990 through 2001. AB - BACKGROUND: Thailand's epidemic of HIV infection, which began in 1988, has primarily involved heterosexual transmission of the virus. This study describes changes in prevalence of HIV and other infectious diseases among blood donors in northern Thailand from 1990 through 2001. STUDY DESIGN AND METHODS: Serologic screening results and demographic data were analyzed from 276,066 donors screened at two blood collection facilities in Chiang Mai, Thailand, from 1990 through 2001. RESULTS: The HIV prevalence peaked in 1991 to 1993 at 4.04 percent and then declined to 0.38 percent in 2001. The overall prevalence of HIV infection was 2.16 percent; HIV prevalence was higher among male (2.24%) than among female (0.64%) donors, in first-time donors, and in replacement volunteer donors. The majority of the donors were men and first-time donors throughout this study. The prevalence of antibodies to syphilis decreased significantly in both men and women. However, the prevalence of antibodies to HCV and HBsAg were stable. CONCLUSIONS: The declining HIV prevalence from 1990 through 2001 among blood donors in two large blood banks in northern Thailand indicates significant progress toward recruitment of a safer donor population in a developing country despite a major HIV and AIDS epidemic involving the general population. PMID- 12757524 TI - Circulatory arrest during PBPC apheresis in an unrelated donor. AB - BACKGROUND: Nowadays, the collection of PBPCs by apheresis from healthy donors is a routine method. The mobilization with rHu G-CSF and the apheresis procedures are usually well tolerated without severe side effects. STUDY DESIGN AND METHODS: We report a severe complication in a 41-year-old unrelated female donor who was allowed to donate PBPCs and was mobilized with 10 microg of G-CSF per kg per day. During PBPC apheresis, she experienced a circulatory arrest after 132 minutes and processing of 7078 mL of blood (twice the donor's blood volume). RESULTS: Immediate cardiopulmonary resuscitation restored sinus rhythm and regulatory respiration without sequelae. Subsequent cardiologic examinations (heart catheterization, electrophysiologic testing, tilting table test) resulted in the diagnosis of a neurocardiogenic syncope. Other cardiac or circulatory disorders could be excluded. The implantation of a cardiac pacemaker was recommended to the donor. The 4-year-old recipient was successfully transplanted with the partial product collected until the arrest occurred. The patient received a total of 2.54 x 106 CD34+ cells per kg of body weight. CONCLUSION: After exclusion of other cardiac diseases, the diagnosed neurocardiogenic syncope probably induced the circulatory arrest during apheresis rather than the administration of G-CSF. PMID- 12757525 TI - Methodologic issues in the use of bleeding as an outcome in transfusion medicine studies. AB - BACKGROUND: Prophylactic platelet transfusions are given to thrombocytopenic patients to prevent bleeding. The benefit of platelet transfusions has frequently been assessed by measuring the count increment; however, more recently, an assessment of bleeding has been used because it is a more clinically relevant outcome measure. The purpose of this study was to identify platelet transfusion trigger studies that used bleeding as an outcome measure, compare and contrast methods used to document bleeding and analyze bleeding outcomes, and identify and discuss methodologic issues to consider when bleeding is used as a study outcome. STUDY DESIGN AND METHODS: A systematic search to identify platelet transfusion trigger studies was performed. Relevant articles were reviewed to identify how bleeding data was captured and analyzed, and methodologic considerations were identified. RESULTS: Seven articles meeting the predefined entry criteria were identified. Methods used to document bleeding included chart review and clinical assessment. The frequency of assessment and the type of personnel performing the assessment were variable. Four approaches to analysis were identified: descriptive; comparison of the proportions of patients having at least one bleed; comparison of patient days with bleeding expressed as a proportion of the total days at risk of bleeding; and time-to-event (first bleed) analysis. CONCLUSION: Methodologic issues for consideration when designing a clinical study with bleeding as the outcome measure included approaches to minimize bias in the documentation and classification of bleeding and selection of an analysis approach that is appropriate to the question being asked. The need for development of a valid and reliable bleeding scale was also identified. PMID- 12757526 TI - Acute and delayed hemolytic transfusion reactions secondary to HLA alloimmunization. AB - BACKGROUND: HLA antibodies may be directed against HLA antigens on RBCs, but these antibodies are generally not considered to be clinically significant in transfusion practice. A case of a multiparous woman who had hemolytic transfusion reactions due to HLA-related Bg antibodies is reported. CASE REPORT: A 37-year old woman was admitted with anemia. No unexpected RBC antibodies were identified. Two group O, D+ RBC units were transfused. Ten days later she returned with hemolysis and anemia. Two more RBC units were ordered, no unexpected RBC antibodies were identified, and two crossmatch-compatible units were issued. During the transfusion, the patient developed symptoms of an acute reaction, and the posttransfusion sample showed evidence of intravascular hemolysis. RESULTS: Repeat RBC antibody screen showed anti-Bg. HLA antibody screen identified anti HLA-A2, A28, B7, B7 cross-reactive group (CREG). The two RBC units from the first transfusion episode and one RBC unit from the second transfusion episode were HLA incompatible with the patient. No other cause for the hemolytic reactions was identified. The patient was later successfully transfused with one RBC unit from an HLA-compatible donor. CONCLUSION: HLA antibodies should be considered in patients with hemolytic transfusion reactions when RBC-specific antibodies are not found to be the etiology. PMID- 12757527 TI - Production of recombinant murine-human chimeric IgM and IgG anti-Js(b) for use in the clinical laboratory. AB - BACKGROUND: Directly agglutinating MoAbs are more useful than IgG MoAbs of murine origin for typing RBCs from donors and patients. The molecular manipulation and conversion of a murine IgG MoAb into mouse- human chimeric IgM and IgG antibodies are described. STUDY DESIGN AND METHODS: cDNA encoding the variable heavy- and light-chain genes of a murine hybridoma anti-Jsb cell line (MIMA-8) were cloned into human IgM or IgG expression vectors, which were then separately stably transfected into SP2/0-Ag14 B-cells. The secreted antibodies were screened by ELISA and analyzed by flow cytometry and hemagglutination. RESULTS: Forty percent (16 of 40) of the stable clones secreted IgM and 66 percent (12 of 18) of the stable clones secreted IgG. The chimeric IgM from the highest expressing clone reacted 4+ in LISS at room temperature. The chimeric IgG from one clone reacted 4+ by the IAT, resembling the specificity of the original murine antibody. Both manipulated MoAbs reacted specifically with RBCs as assessed by flow cytometry. CONCLUSION: Human-mouse chimeric IgM and IgG from a murine IgG MoAb anti-Jsb has been successfully engineered for use in the clinical laboratory. This approach can potentially be used to manipulate other murine MoAbs to blood group antigens into more clinically useful human isotypes. PMID- 12757529 TI - Differences in the perception of blood transfusion risk between laypeople and physicians. AB - BACKGROUND: There is little objective evidence to support the commonly held belief that laypeople perceive blood transfusion risk differently from physicians. Acknowledging and characterizing such differences may improve risk communication. The objective of this study was to characterize how laypeople and physicians perceive the risks of blood transfusion in comparison with a wide variety of other hazards. STUDY DESIGN AND METHODS: A total of 161 laypeople and 91 physicians and medical trainees were surveyed in Kingston, Ontario, between March and August 2000. The perceived riskiness and other qualitative characteristics of blood transfusion and 9 other hazards were measured by psychometric scaling and principal components analysis. RESULTS: The overall return rate was 100 percent, with 86 percent of surveys having no missing responses. Physicians perceived the risks of blood transfusion and most other hazards to be less dreaded and severe, but also less understood and controllable than laypeople. CONCLUSION: Laypeople and physicians perceive risk differently for blood transfusion, but this perceptual gap between the groups for blood transfusion may be representative of a more generalized phenomenon that spans different types of hazards, both medical and nonmedical. Awareness of such differences may facilitate risk communication and shared decision making between physicians and their patients. PMID- 12757528 TI - HLA and RBC immunization after filtered and buffy coat-depleted blood transfusion in cardiac surgery: a randomized controlled trial. AB - BACKGROUND: WBC reduction of all blood components is being introduced in many countries. Prevention of immunologic side effects of transfusions is part of the motivation. To compare the immunogenicity of before- or after-storage WBC-reduced RBCs with RBCs without buffy coat, a randomized clinical trial was performed. STUDY DESIGN AND METHODS: Cardiac surgery patients were randomly assigned to receive either RBCs without buffy coat (PCs), WBC-reduced RBCs that were filtered before storage (FFs), or WBC-reduced RBCs that were filtered after storage (SFs). Serum samples for antibody analyses were collected before and after surgery. RESULTS: Sera of 404 patients were tested. Of the 317 patients with negative preoperative screening, 12.6 percent developed anti-WBC antibodies (PC, 14.5%; FF, 9.6%; SF, 13.3%). Of the 87 patients with preoperative anti-WBC antibodies, 28.7 percent showed a marked increase in panel reactivity (PC, 31.3%; FF, 29.0%; SF, 25.0%). ELISA showed the newly formed antibodies to be of IgG class and directed against HLA class I in more than 90 percent of the samples tested. Newly formed anti-RBC antibodies appeared in 5.3 percent (PC, 7.1%; FF, 3.4%; SF 5.4%). Alloimmunization against WBCs and RBCs was strongly correlated (p < 0.01). The differences in newly formed anti-WBC antibodies and anti-RBC antibodies between the trial arms did not show significance. CONCLUSION: Buffy coat removal, and additional WBC reduction by filtration, either before or after storage, result in similar posttransfusion alloimmunization frequencies after a single transfusion event with multiple RBCs. PMID- 12757530 TI - Evaluation of a visual risk communication tool: effects on knowledge and perception of blood transfusion risk. AB - BACKGROUND: Effective risk communication in transfusion medicine is important for health-care consumers, but understanding the numerical magnitude of risks can be difficult. The objective of this study was to determine the effect of a visual risk communication tool on the knowledge and perception of transfusion risk. STUDY DESIGN AND METHODS: Laypeople were randomly assigned to receive transfusion risk information with either a written or a visual presentation format for communicating and comparing the probabilities of transfusion risks relative to other hazards. Knowledge of transfusion risk was ascertained with a multiple choice quiz and risk perception was ascertained by psychometric scaling and principal components analysis. RESULTS: Two-hundred subjects were recruited and randomly assigned. Risk communication with both written and visual presentation formats increased knowledge of transfusion risk and decreased the perceived dread and severity of transfusion risk. Neither format changed the perceived knowledge and control of transfusion risk, nor the perceived benefit of transfusion. No differences in knowledge or risk perception outcomes were detected between the groups randomly assigned to written or visual presentation formats. CONCLUSION: Risk communication that incorporates risk comparisons in either written or visual presentation formats can improve knowledge and reduce the perception of transfusion risk in laypeople. PMID- 12757532 TI - Retrospective targeted HCV lookback using centralized contracted notification service. AB - BACKGROUND: Mandated HCV 2.0 lookback significantly challenged the human and financial resources available to the six Los Angeles County Department of Health Services (DHS) hospital blood banks. To comply with FDA requirements, DHS developed a centralized process that utilized the services of a contract vendor for performing HCV lookback. STUDY DESIGN AND METHOD: A DHS public health nurse acted as HCV lookback coordinator and as liaison with the vendor. Cases were electronically forwarded to the vendor, who then reviewed the patient's chart to obtain information necessary for tracking the patient through commercial databases. The vendor was responsible for notifying the recipient for pretest counseling and for providing documentation of all efforts. RESULTS: In total, 411 recipients were identified, of which, 168 cases were completed by the hospitals, 243 were forwarded to the vendor, 50 percent were deceased, 21 percent were contacted, and 28 percent could not be contacted, and 1 percent could not be located on any databases. The vendor contacted 35 of 66 cases that the hospitals had unsuccessfully attempted to contact. Of the cases forwarded to the vendor, 82 percent were completed on time. Delays were attributable to the vendor in less than 2 percent of cases. The cost per case forwarded to the vendor was 322 US dollars. CONCLUSION: Utilizing a professional service to conduct HCV recipient notifications is an effective strategy for meeting FDA deadlines and for freeing transfusion service staff to perform critical functions. It remains to be seen whether such strategies offer a cost savings. PMID- 12757531 TI - Comparative sensitivity of HBV NATs and HBsAg assays for detection of acute HBV infection. AB - BACKGROUND: A study was designed to estimate relative analytic sensitivity and window-period (WP) closure and to project incremental yield of newer HBsAg tests, pooled-sample NAT, and single-sample NAT, compared to currently licensed HBsAg tests. STUDY DESIGN AND METHODS: HBV DNA and HBsAg test results for 23 HBV seroconversion (SC) panels were first analyzed to construct a model of primary HBV viremia. One-hundred representative samples were then selected from 10 panels and coded with 28 analytical controls. All 128 samples were tested by seven HBsAg tests and by four pooled-sample and three single-sample NAT assay formats. Results were analyzed to obtain differential times to HBV detection and combined with HBV incidence rates to project comparative yields. RESULTS: HBV doubling time during the ramp-up phase was estimated at 2.56 days. HBsAg concentrations at cutoff for new tests ranged from 0.07 to 0.12 ng per mL, compared with 0.13 to 0.62 ng per mL for licensed tests. Estimated viral load at cutoff ranged from 102 to 267 IU per mL for new tests and from 363 to 1069 IU per mL for licensed tests. HBsAg tests detected 31 to 63 percent of early ramp-up phase samples in the 100 member seroconversion panel study, while pooled-sample NAT detected 55 to 71 percent and single-sample NAT, 82 to 99 percent. Compared with currently licensed HBsAg assays, newer HBsAg assays would reduce the WP by 2 to 9 days; pooled sample NAT would reduce the WP by 9 to 11 days; and single-sample NAT would reduce the WP by 25 to 36 days. CONCLUSION: Newer HBsAg tests would be expected to detect an additional 15 to 21 infected units per 107 donations, compared to licensed HBsAg tests. Sensitivity, WP closure, and yield projections for newer HBsAg assays and pooled-sample NAT are comparable. Single-sample NAT would increase yield by 13 to 15 units per 107 donations over pooled-sample NAT and newer HBsAg assays and by 35 to 50 units per 107 donations over currently licensed HBsAg assays. PMID- 12757533 TI - Cord blood processing with an automated and functionally closed system. AB - BACKGROUND: Umbilical cord blood processing with standard centrifugation techniques is performed in open systems and results in varying cell and volume recoveries. STUDY DESIGN AND METHODS: Forty umbilical cord blood donations were randomly assigned to processing either with a microprocessor-controlled cell separator equipped with closed disposables or with a manual separation procedure in blood bags. The collection efficiency of nucleated cells, MNCs, RBCs, and CD34+ cells and the processing time were analyzed. RESULTS: Using the cell processor, mean collection efficiencies were 78.6 +/- 24.9 percent for nucleated cells, 77.4 +/- 27.8 percent for MNCs, 55.5 +/- 14.6 percent for RBCs, and 83.6 +/- 32.5 percent for CD34+ cells, while they were 73.1 +/- 13.2 percent for nucleated cells, 78.1 +/- 14.9 percent for MNCs, 26.0 +/- 12.2 percent for RBCs, and 77.0 +/- 17.6 percent for CD34+ cells when using the standard centrifugation technique. The processing time was about 20 minutes for automated processing and 60 to 80 minutes for the standard centrifugation technique. CONCLUSION: Using the new cell processor, the collection efficiencies for nucleated cells, MNCs, and CD34+ cells are similar to those obtained by established centrifugation techniques while the RBC reduction is less effective. The main advantages of the new systems are the closed system, the more standardized processing procedure, and a significantly shorter processing time. PMID- 12757534 TI - Collection of MNCs and progenitor cells by two separators for PBPC transplantation: a randomized crossover trial. AB - BACKGROUND: Efficient collection of progenitor cells is essential for PBPC transplantation. Two apheresis machines (Amicus, Baxter Healthcare; and Spectra, Gambro BCT, software version 4.7) were compared prospectively by a crossover trial. STUDY DESIGN AND METHODS: Apheresis collections were performed for two consecutive days on patients for autologous and donors for allogeneic PBPC transplantation. The patients and donors, receiving a G-CSF, were randomized into two groups. In Group I, PBPCs were collected by the Amicus on the 1st day and the Spectra on the 2nd day, and the reverse order was used with Group II. A total of 60 apheresis procedures of 30 (16 in Group I and 14 in Group II) among 40 patients and donors enrolled were performed and evaluated. RESULTS: The nucleated cell counts, MNC counts, CD34+ PBPC counts, and amounts of CFU-GM collected per procedure were similar with the Amicus and the Spectra. On the other hand, the decrease of peripheral blood platelet counts of patients and donors was more prominent from using Spectra than Amicus (p < 0.0001). Components collected by the Amicus had fewer platelets than those collected by the Spectra (p < 0.0001). The efficiencies of collecting nucleated cells, MNCs, and CD34+ PBPCs were not different between the machines (p > 0.05). However, the efficiency of collecting platelets was significantly higher with Spectra than with Amicus (p < 0.0001). The Amicus took longer than the Spectra to process the same volume (p < 0.05). CONCLUSION: Amicus is superior to Spectra in avoiding apheresis-induced thrombocytopenia caused by platelets contaminating the collected samples. Therefore, the Amicus is useful for patients with thrombocytopenia or with a less than-normal platelet count. PMID- 12757535 TI - Therapeutic apheresis: a summary of current indication categories endorsed by the AABB and the American Society for Apheresis. PMID- 12757536 TI - Transfusion medicine: discipline with a future. PMID- 12757537 TI - Platelet transfusion and thrombotic thrombocytopenic purpura. PMID- 12757539 TI - Risk of transfusion-transmitted infections by NAT-negative blood. PMID- 12757541 TI - Effect of universal WBC reduction. PMID- 12757543 TI - HCV NAT in Austria. PMID- 12757545 TI - Enzyme treatment of platelets for antibody detection. PMID- 12757547 TI - Traceability of inbred and crossbred Cinta Senese pigs by evaluating the oxidative stress. AB - Previous studies on the oxidative stress in swine indicated a strong link between the values of reactive oxygen metabolites (ROMs), the subsequent antioxidant adaptive response (OXY) and the genetic selection. Such findings, mainly related to a cardiovascular inadequacy in lean, large muscle blocks and fast growing breeds, is associated with specific metabolic diseases such as porcine stress syndrome and mulberry heart disease. In this study, we investigated the oxidative stress parameters to trace the genetics of Cinta Senese (CS) pigs, a historical breed free-range reared in Siena countryside. Sera from CS (n = 24) and Large White x CS (LW x CS) (n = 24) groups around 120 kg body weight fed the same diet were sampled at slaughter. Sera from wild boars (WB) (n = 24) hunted in the same district were also considered. ROMs and OXY were evaluated in the three groups of swine. Significant differences by one-way anova (P < 0.05) between groups were found for both procedures. ROM levels were lower in WB (9.79 +/- 5.76 mm H2O2) and CS (18.02 +/- 7.42 mm H2O2), and highest in LW x CS (42.78 +/- 8.61 mm H2O2). OXY values ranged from 271.37 +/- 50.90 microm neutralized HOCl (WB) to 343.21 +/ 57.45 microm neutralized HOCl (LW x CS). Results indicated that the evaluation of the oxidative stress can effectively trace the CS pigs, thus aiding in preserving the overall specific traits of such a historical animal. PMID- 12757548 TI - Effects of hypothyroidism on the skeletal muscle blood flow response to contractions. AB - Hypothyroidism is associated with impaired blood flow to skeletal muscle under whole body exercise conditions. It is unclear whether poor cardiac and/or vascular function account for blunted muscle blood flow. Our experiment isolated a small group of hindlimb muscles and simulated exercise via tetanic contractions. We hypothesized that muscle blood flow would be attenuated in hypothyroid rats (HYPO) compared with euthyroid rats (EUT). Rats were made hypothyroid by mixing propylthiouracil in their drinking water (2.35 x 10-3 mol/l). Treatment efficacy was evidenced by lower serum T3 concentrations and resting heart rates in HYPO (both P<0.05). In the experimental preparation, isometric contractions of the lower right hindlimb muscles at a rate of 30 tetani/min were induced via sciatic nerve stimulation. Regional blood flows were determined by the radiolabelled microsphere method at three time points: rest, 2 min of contractions and 10 min of contractions. Muscle blood flow generally increased from rest ( approximately 5-10 ml/min per 100 g) through contractions for both groups. Further, blood flow during contractions did not differ between groups for any muscle (eg. red section of gastrocnemius muscle; EUT, 59.9 +/- 14.1; HYPO, 61.1 +/- 15.0; NS between groups). These findings indicate that hypothyroidism does not significantly impair skeletal muscle blood flow when only a small muscle mass is contracting. Our findings suggest that impaired blood flow under whole body exercise is accounted for by inadequate cardiac function rather than abnormal vascular function. PMID- 12757549 TI - Characteristics of lactoferrin receptor in bovine intestine: higher binding activity to the epithelium overlying Peyer's patches. AB - Several lines of evidence have recently demonstrated the occurrence of specific lactoferrin (Lf) receptors in different cells. We report here, for the first time, the characteristics of binding, and distribution of Lf receptors in the bovine intestinal tract with special emphasis on the epithelium overlying Peyer's patches (EOPP). Brush-border membrane vesicles (BBMV) were prepared from the mucosa of duodenum, jejunum, ileum, colon, EOPP in jejunum and EOPP in ileum. Receptor binding assays were carried out using 125I-labelled bovine Lf. Specific and saturable Lf receptors were found in BBMV of all the intestinal segments examined. Non-linear regression and Scatchard plot analyses clearly revealed that EOPP had the highest binding maximal (Bmax), and lowest in colon. The maximum dissociation constant (Kd) 3.74 microm was in the ileum. We found that bovine transferrin competed with Lf for the same binding site of receptors. In contrast, no binding of bovine serum albumin occurred. It was concluded that Lf receptors in the mucosal lining are attributable to mediate multifunctional activities of Lf in the gut, especially in the EOPP. PMID- 12757551 TI - Immunohistochemical characteristics of canine aortic and carotid body tumours. AB - In an immunohistochemical study of 25 canine chemodectomas, 17 tumours were stained with antisera to neurone specific enolase and the same number were stained for synaptophysin; a single tumour was stained for S100. Staining for Ki 67 occurred in 18 cases; the Ki-67-labelling index and the intensity of immunostaining was increased in more pleomorphic and malignant tumours, as assessed on histological grounds. Immunohistochemistry did not aid in recognition of less well-differentiated tumours. PMID- 12757550 TI - Effects of prenatal stress on corticosteroid receptors and monoamine concentrations in limbic areas of suckling piglets (Sus scrofa) at different ages. AB - The present study was conducted in order to reveal the effects of prenatal stress on the central stress regulation in domestic pigs by measuring changes in corticosteroid receptor binding and monoamine concentrations in different limbic brain regions. Pregnant sows were subjected to a restraint stress for 5 min daily during the last 5 weeks of gestation. Maternal stress resulted in a significantly higher number of glucocorticoid receptors in the hippocampus, but decreased glucocorticoid receptors in the hypothalamus of the offspring at the first postnatal day. No alterations of hippocampal mineralocorticoid receptors were found. There was also no significant effect of prenatal stress on the brain monoamine concentrations. Prenatally stressed piglets showed lower basal plasma cortisol and increased corticosteroid binding globulin concentrations at the third postnatal day indicating decreased free cortisol concentrations after birth. Morbidity and mortality during the suckling period were significantly increased in prenatally stressed litters, as shown by a higher frequency of diseased and died piglets per litter. In conclusion, the results indicate that in pigs restraint stress during late gestation affects the ontogeny of the foetal neuroendocrine feedback system with consequences for the regulation of the hypothalamo-pituitary-adrenal function and the vitality of the offspring. PMID- 12757552 TI - Polioencephalomalacia in captive harbour seals (Phoca vitulina). AB - In a colony of 11 harbour seals (Phoca vitulina Linne 1758) two episodes of central nervous disorders occurred within 2 years causing fatalities in seven adult animals. Clinical signs comprised dyspnoea, anorexia, apathy, incoordination and lateral recumbency. Vitamin B complex therapy was successful once. Pathomorphological examination of seven carcasses revealed acute and subacute malacia of the cerebellar grey matter. Additional acute malacic lesions located in the cerebral cortices and basal ganglia were observed. Mesencephalic nuclei were less severely affected and displayed acute changes. Despite intense search for environmental toxins and infectious agents, the cause of the fatalities remained undetermined. However, the type and pattern of the lesions are most suggestive of a thiamine deficiency. PMID- 12757553 TI - Extranodal lymphoblastic lymphoma of suspected B-cell lineage in the gingiva of a racehorse, accompanied by mandibular osteolysis. AB - A mass developed in the mandibular gingiva of a thoroughbred racehorse. When the horse could no longer eat unassisted, it was killed and immediately autopsied. Macroscopically, the mandible exhibited extensive osteolysis, with only a small amount of bone remaining around the tooth roots. The cut surface of the mass around the mandible consisted of neoplastic medullary tissue, in which osteogenesis was observed. The medullary tissue was composed of pleomorphic medium-sized to large cells, interlaced by collagen bundles. These cells had large, pale, round or ovoid, sometimes cleaved nuclei, with one or two prominent nucleoli. Mitoses were numerous. Electron microscopy showed that the cells in the medullary tissues were similar in shape to undifferentiated lymphocytes. Immunohistochemically, these cells were positive for B-cell associated antigen in the pre-B-cell stage. Our findings suggest that the horse had extranodal lymphoblastic lymphoma of suspected B-cell lineage, possibly originating from the lymphatic system of the gingiva. We consider that the osteolysis resulted from activation of osteoclasts by proliferation of the tumour cells. PMID- 12757554 TI - Induction of apoptosis by canine natural killer cells. AB - Besides a secretory pathway of canine natural killer (NK) cells, which results in necrosis of the target cell, a second pathway was demonstrated, which results in apoptosis of the target cell. Comparing the Chromium Release Assay (CRA) and the Rose Bengal Assay (RBA) for quantification of in vitro canine NK cell activity, a constant 10% higher NK cell activity was found in the RBA compared with the CRA. To find out the mechanism responsible for the different results of both tests, morphological studies of in vitro canine NK cell activity against epithelial and mesenchymal allogenic target cell lines were performed. Most target cells were undergoing necrosis as a result of NK cell killing, which was evidenced by transmission electron microscopy. However, besides necrotic target cells, shrunken target cells with dense cytoplasm, fragmented nuclei and disruption into membrane-bound bodies were detected, which are known as signs of apoptosis. Additionally, using the terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling (TUNEL) method, 13-23% of target cells presented a positive staining, indicative of apoptosis. These findings give evidence for the ability of canine NK cells to kill their target cells via two different pathways, which results either in apoptosis or necrosis. PMID- 12757555 TI - Xylazine does not induce retrograde flow of spermatozoa into the urinary bladder of sexually rested boars. AB - The effect of xylazine on the retrograde flow of spermatozoa from their storage sites in the epididymides and vasa deferentia into the urinary bladder of sexually rested boars was examined. The bladder of four boars was evacuated through a surgically implanted urinary catheter and the urine was examined for the presence of spermatozoa. Boars were then given an injection of 2.2 mg of xylazine per kilogram of body weight and, immediately thereafter, 500 ml of saline was infused into the urinary bladder. Approximately 50 ml of the post treatment mixture of urine and saline, referred to as 'urine', was collected through the catheter at 5, 10, 15, 20, 30 and 45 min after the injection of xylazine, and examined immediately for the presence and motility of spermatozoa. At 60 min, the urinary bladder was evacuated and the remaining 'urine' was examined for the presence and motility of spermatozoa. None of the pre-xylazine urine and post-xylazine fractions of 'urine' had motile spermatozoa and xylazine did not increase (P > 0.1) the concentration and the number of spermatozoa in the post-treatment 'urine'. Thus, in contrast to other species, xylazine does not induce retrograde flow of spermatozoa into the urinary bladder of boars. PMID- 12757556 TI - Evaluation of a commercially available enzyme-linked immunosorbent assay (ELISA) for the determination of C-reactive protein in canine serum. AB - The purpose of this study was to evaluate a commercially available enzyme-linked immunosorbent assay for determination of canine serum C-reactive protein (CRP). The concentration of CRP could be determined accurately and the intra- and inter assay coefficients of variation were in the range of 6.9-10.1 and 7.5-29.0%, respectively. This level of imprecision between runs is usually considered unacceptable for diagnostic purposes, but the overall results indicated that the assay was useful in differentiating dogs suffering from infections, from dogs suffering from various other diseases (neoplastic diseases, endocrine/metabolic disorders), and healthy dogs. The assay was also able to detect dynamic changes of CRP during development and after cessation of spontaneous occurring inflammatory stimuli in two clinical cases. PMID- 12757558 TI - Efficacy of donepezil on maintenance of activities of daily living in patients with moderate to severe Alzheimer's disease and the effect on caregiver burden. AB - OBJECTIVES: This study investigated the efficacy of donepezil treatment on activities of daily living (ADLs) and social functioning in patients with moderate to severe Alzheimer's disease (AD) and the possible benefits of this treatment on caregiving time and stress levels. DESIGN: Randomized, double-blind, placebo-controlled, multinational study. SETTING: Patients resided in the community or in assisted living facilities who did not require skilled 24-hour nursing care. PARTICIPANTS: Two hundred ninety patients with moderate to severe AD (baseline standardized Mini-Mental State Examination score of 5-17). INTERVENTION: Donepezil (5 mg/d for 4 weeks and 10 mg/d per clinician's judgment thereafter) or placebo for 24 weeks. MEASUREMENTS: ADLs were assessed using the Disability Assessment for Dementia (DAD), the modified instrumental activities of daily living (IADL) scale (IADL+), and the modified Physical Self Maintenance Scale (PSMS+). Caregiver time spent assisting patients with basic and instrumental ADLs was recorded as part of the IADL+ and PSMS+ scales. Patients' social behavior was evaluated through the use of a caregiver diary that captured observations of patients' motivation, interactions, and engagement. Caregivers were evaluated for their levels of caregiver stress with a modified, multiple item Caregiver Stress Scale (CSS). For each outcome measure, the mean change from baseline at Week 24 using a last observation carried forward (LOCF) analysis was calculated. RESULTS: IADL+ and PSMS+ mean change from baseline scores for donepezil-treated patients showed a slower decline during the study than placebo treated patients (Week 24 LOCF mean treatment differences: IADL+ = 6.83, P <.0001; PSMS+ = 1.32, P =.0015). Significant differences between the groups in favor of donepezil were observed on the DAD for instrumental and basic ADLs and on the three components required for the completion of each ADL: initiation, planning and organization, and effective performance. At baseline, caregivers of patients treated with donepezil (n = 141) did not differ significantly from caregivers of patients treated with placebo (n = 146) with respect to demographics or mean total scores on the CSS. At Week 24 LOCF, the overall distribution of caregiver ratings on each of the three caregiver diary items favored donepezil-treated patients over placebo-treated patients (P <.005). At Week 24 LOCF, mean change from baseline scores for CSS total and individual domain scores (all domains except caregiving competence, personal gain, and management of distress) were better for caregivers of donepezil-treated patients than for those of placebo-treated patients (CSS total, mean treatment difference = 1.82). Caregivers of donepezil-treated patients reported spending less time assisting with ADLs than caregivers of placebo-treated patients (mean difference = 52.4 min/d). CONCLUSION: Donepezil demonstrated a significantly slower decline than placebo in instrumental and basic ADLs in these patients with moderate to severe AD. The ADL benefits in AD patients treated with donepezil were associated with less caregiving time and lower levels of caregiver stress. PMID- 12757559 TI - Functional status and health outcomes in older americans with diabetes mellitus. AB - OBJECTIVES: To determine how baseline functional status affects health outcomes in older adults with diabetes mellitus (DM). DESIGN: Nationally representative cross-sectional and longitudinal health interview survey. SETTING: Waves I (1993) and II (1995) of the Assets and Health Dynamics of the Oldest Old Survey (AHEAD) in the United States. PARTICIPANTS: AHEAD respondents aged 70 and older (n = 7,447, including 995 with DM). MEASUREMENTS: At baseline, the entire sample was divided into three groups: high-functioning group, defined as having no physical limitations or instrumental activities of daily living/activities of daily living (IADL/ADL) disabilities (39%); low-functioning group, having three or more limitations or IADL/ADL disabilities (24%); and intermediate-functioning group, those in the middle (36%). Older adults with and without DM, within each of the functioning groups, were compared at 2-year follow-up with respect to demographic characteristics, weight/body mass index, baseline and incident chronic diseases and conditions, and follow-up functioning. RESULTS: Of people aged 70 and older, 28% with DM and 41% without were high functioning; 38% with DM and 22% without were low functioning (both P <.001). High-functioning people with DM remained high functioning at 2 years but had a significantly higher incidence of heart disease and mortality than high-functioning people without DM. Low-functioning people with DM were significantly more likely to have vascular comorbidities at baseline than low-functioning people without DM, but their 2-year outcomes were similar. The intermediate-functioning group showed the most differences between those with and without DM; those with DM were significantly (P <.01) more likely to have baseline vascular disease, low cognitive performance, increased incident vascular disease, and significantly worse 2-year functioning and to have experienced falls (P <.001). CONCLUSION: Differences in baseline functional status in older adults with DM were associated with outcome differences. High functioning older people with DM tended to remain high functioning but demonstrated significantly higher incidence of heart disease and mortality than those without DM, whereas low-functioning people with and without DM had similar outcomes. However, intermediate-functioning older diabetics had worse health and functioning outcomes than a similarly impaired group without DM. DM management adjusted to functional status can potentially address the most-relevant outcomes in the heterogeneous older population with DM. PMID- 12757560 TI - The prognostic significance of subsyndromal delirium in elderly medical inpatients. AB - OBJECTIVES: To determine the prognostic significance of subsyndromal delirium (SSD) presentations. DESIGN: Cohort study. SETTING: University-affiliated primary acute care hospital. PARTICIPANTS: One hundred sixty-four elderly medical inpatients who did not meet Diagnostic and Statistical Manual of Mental Disorders, Third Edition, Revised (DSM-III-R) criteria for delirium during the first week after admission were classified into three mutually exclusive groups. The first group, prevalent SSD, included those who had two or more of four core symptoms of delirium (clouding of consciousness, inattention, disorientation, perceptual disturbances) at admission. The second group, incident SSD, included those who did not meet criteria for prevalent SSD but displayed one or more new core symptoms during the week after admission. The third group had no prevalent or incident SSD. The three groups were followed up at 2, 6, and 12 months. MEASUREMENTS: Outcomes (length of stay, symptoms of delirium (Delirium index), cognitive (Mini-Mental State Examination) and functional status (instrumental activities of daily living), and mortality) were compared using univariate techniques and multivariate regression models that adjusted for age, sex, marital status, living arrangements before admission, comorbidity, clinical and physiological severity of illness, and dementia status and severity. RESULTS: Patients with prevalent SSD had longer acute care hospital stay, increased postdischarge mortality, more symptoms of delirium, and a lower cognitive and functional level at follow-up than patients with no SSD. Most of the findings for incident SSD were similar but not statistically significant. Patients with prevalent or incident SSD had risk factors for DSM-defined delirium. CONCLUSION: SSD in elderly medical inpatients appears to be a clinically important syndrome that falls on a continuum between no symptoms and DSM-defined delirium. PMID- 12757561 TI - Influenza- and respiratory syncytial virus-associated morbidity and mortality in the nursing home population. AB - OBJECTIVES: To estimate winter viral-related morbidity and mortality in Tennessee nursing home residents during 4 consecutive years. DESIGN: A retrospective cohort study. SETTING: Three hundred eighty-one Tennessee nursing homes. PARTICIPANTS: Nursing home residents. MEASUREMENTS: Viral surveillance data were used to define three seasons: influenza (influenza and respiratory syncytial virus (RSV) cocirculating), RSV (RSV alone circulating), and non winter-viral (neither virus circulating). Adjusted seasonal differences in rates of cardiopulmonary hospitalizations, antibiotic prescriptions, and deaths during these three seasons were calculated to estimate annual hospitalizations, courses of antibiotics, and deaths attributable to influenza and RSV from 1995 to 1999. RESULTS: Nursing home residents had 81,885 person-years of follow-up. In the 63% of residents with comorbid conditions that increase influenza morbidity, influenza infection contributed to an estimated average of 28 hospitalizations, 147 courses of antibiotics, and 15 deaths per 1,000 persons annually. Similarly, RSV accounted for an annual average of 15 hospitalizations, 76 courses of antibiotics, and 17 deaths per 1,000 persons. Influenza and RSV accounted for 7% of cardiopulmonary hospitalizations and 9% of total deaths in high-risk residents during the 4 study years. Absolute morbidity and mortality were lower in residents without identified comorbid conditions but accounted for 15% of hospitalizations and 14% of deaths. These estimates depend on the assumption that morbidity and mortality from other respiratory viruses were distributed evenly between the three defined seasons. CONCLUSION: Influenza and RSV substantially increased hospitalization rates, antibiotic use, and deaths in elderly nursing home residents each winter. These data should encourage persistent efforts toward disease prevention, and thoughtful study of vaccine development and delivery, diagnostic tools, and methods of prophylaxis and therapy. PMID- 12757562 TI - Prevalence of small bowel bacterial overgrowth and its association with nutrition intake in nonhospitalized older adults. AB - OBJECTIVES: To determine the prevalence of small bowel bacterial overgrowth (SBBO) in older adults and to assess whether SBBO is associated with abdominal complaints and nutrient intake. DESIGN: Cross-sectional survey. SETTING: Eight senior residence sites in Stuttgart, Germany. PARTICIPANTS: Older adults living independently in senior residence houses. MEASUREMENTS: The prevalence of SBBO was measured in 328 subjects, of whom 294 were aged 61 and older, by measuring hydrogen concentration (parts per million; ppm) in exhaled air after ingestion of 50 g glucose. Anthropometric data were obtained and nutritional status was recorded with a computer-aided diet history. RESULTS: The prevalence of a positive hydrogen breath test (>10 ppm increase) was 15.6% in older adults, compared with 5.9% in subjects aged 24 to 59. The intake of inhibitors of gastric acid production contributed significantly to the high prevalence of a positive breath test in older adults, which was associated with lower body weight, lower body mass index, lower plasma albumin concentration, and higher prevalence of diarrhea. Subjects with a positive hydrogen breath test consumed significantly less fiber, folic acid, and vitamins B2 and B6 than those without. No difference was observed in the intake of energy, protein, fat, or carbohydrates. CONCLUSION: Prevalence of SBBO is associated with reduced body weight, which is paralleled by reduced intake of several micronutrients. Malabsorption resulting from diarrhea might be an aggravating factor contributing to weight loss in these subjects. PMID- 12757563 TI - Hypertension in older urban Native-American primary care patients. AB - OBJECTIVES: To examine hypertension and its management in a population of older urban American Indians and Alaska Natives (AI/ANs). DESIGN: Retrospective cohort study using medical record review. SETTING: Urban health clinic serving predominantly AI/ANs in the Pacific Northwest. PARTICIPANTS: Five hundred twenty four AI/ANs aged 50 and older seen between 1994 and 1995. MEASUREMENTS: Frequency of diagnosed hypertension, undiagnosed hypertension, comorbid conditions, hypertension treatment, control, and quality of care. RESULTS: The prevalence of diagnosed hypertension was 38%, and the prevalence of possible undiagnosed hypertension was 23%. Patients with diagnosed hypertension were more likely to be obese (age-adjusted odds ratio (OR) = 3.5), have diabetes mellitus (DM) (OR = 2.2), depression (OR = 1.7), heart disease (OR = 3.8), or renal disease (OR = 5.6) than patients without hypertension. Undiagnosed hypertension was inversely associated with number of health problems (OR = 0.8). Eighty-one percent of diagnosed patients were treated pharmacologically, but no factors associated with nontreatment were identified. Diuretic and beta-blocker usage was low. Patients with DM used angiotensin-converting enzyme inhibitors more frequently than patients without DM (OR = 2.4). Blood pressure was well controlled in 37%, with men being less well controlled than women (OR = 0.5). Serum cholesterol, creatinine, and retinal screening were performed more often than urinalyses or electrocardiograms. Lifestyle-modification counseling was uncommon. Number of health problems was the most common factor associated with screening tests for end-organ disease. CONCLUSION: Few studies have examined the care of older urban AI/ANs. Improvements are needed in adherence to recommendations for the detection, management, and monitoring of hypertension and its complications in older urban AI/ANs. PMID- 12757564 TI - Self-reported disability and its association with performance-based limitation in elderly men: a comparison of three European countries. AB - OBJECTIVES: To compare self-reported disability and performance-based limitation and their association in elderly men from three European countries. DESIGN: Cross sectional data from a cohort study collected around 1990. SETTING: Three cohorts from Finland, the Netherlands, and Italy. PARTICIPANTS: One thousand one hundred sixty-one men aged 70 and older. MEASUREMENTS: Disability and functional limitation were measured in a standardized way in three countries. Self-reported disability was estimated by questionnaire, assessing three domains of activities of daily living: instrumental activities of daily living, mobility, and activities of daily living (score 0-3). Functional limitation was measured by performance tests (score 0-16), with 0 indicated the healthiest score. RESULTS: Self-reported disability and performance-based limitation scores differed between countries. Mean self-reported disability score was worse in Italy (0.72) and the Netherlands (0.70) than in Finland (0.54). Italian men scored worst on the performance-based tests (mean 4.80 vs 4.04 for Finland and 3.74 for the Netherlands). Differences in self-reported disability remained after adjusting for performance scores: Dutch men reported more disabilities (odds ratio (OR) = 1.66, 95% confidence interval (CI) = 1.23-2.25) than men in Finland (reference group) and Italy (OR = 1.08, 95% CI = 0.77-1.53). Self-reported disability was positively associated with performance-based score (OR = 1.28, 95% CI = 1.21 1.35) and did not differ between countries. CONCLUSION: Cross-cultural variation was noted in self-reported disability adjusted for performance score. These differences may be due to sociocultural and physical environmental factors. Self reported disability was consistently associated with performance-based limitation in Finland, the Netherlands, and Italy. PMID- 12757565 TI - Decisions for hospice care in patients with advanced cancer. AB - OBJECTIVES: To identify factors that may influence the decision of whether to enter a hospice program or to continue with a traditional hospital approach in patients with advanced cancer and to understand their decision-making process. DESIGN: Cross-sectional structured interview. SETTING: One community-based hospice and three university-based teaching hospitals. PARTICIPANTS: Two hundred thirty-four adult patients diagnosed with advanced lung, breast, prostate, or colon cancer with a life expectancy of less than 1 year: 173 hospice patients and 61 nonhospice patients receiving traditional hospital care. MEASUREMENTS: Hospice and nonhospice patients' demographic, clinical, and other patient-related characteristics were compared. Multivariate analysis was then conducted to identify variables associated with the hospice care decision in a logistic regression model. Information sources regarding hospice care and people involved in the hospice decision were identified. RESULTS: Patients receiving hospice care were significantly older (average age 69 vs 65 years, P =.009) and less educated (average 11.9 vs 12.9 years, P =.031) and had more people in their households (average 1.66 vs 1.16 persons, P =.019). Hospice patients had more comorbid conditions (1.30 vs 0.93, P =.035) and worse activities of daily living scores (7.01 vs 6.23, P =.030) than nonhospice patients. Hospice patients were more realistic about their disease course than their nonhospice counterparts. Patients' understanding of their prognoses affected their perceptions of the course of their disease. Hospice patients preferred quality of life to length of life. In the multivariate analysis, lower education level and greater number of people in the household were associated with the decision to enter hospice. A healthcare provider first told most of those who entered hospice about hospice. Families largely made the final decision to enter hospice (42%), followed by patients themselves (28%) and physicians (27%). CONCLUSION: The decision to enter hospice is related to demographic, clinical, and other patient-related characteristics. This study suggests that the decision-making process for hospice care in patients with advanced cancer is multidimensional. The healthcare community may better meet the end-of-life care needs of advanced cancer patients through enhanced communication with patients and families, including providing accurate prognoses and better understanding of patients' preferences and values. PMID- 12757566 TI - How does the timing of hospice referral influence hospice care in the last days of life? AB - OBJECTIVES: To determine factors associated with the type of hospice care received in the last days of life and, in particular, how the timing of referral influences the use of continuous hospice home care and inpatient hospice care. DESIGN: Retrospective cohort study. SETTING: Twenty-one hospice programs across seven states under the ownership of one hospice parent provider. PARTICIPANTS: Hospice patients who were cared for and died between October 1, 1998, and September 30, 1999 (N = 28,747). MEASUREMENTS: Patient sociodemographic and clinical data were merged with use data from the provider's centralized information system to examine the factors associated with the differing levels of hospice care in the last week of life. In the last days of life, patients were classified as having received routine hospice home care only, having received continuous hospice home care, or having died in an inpatient hospice bed. RESULTS: Twenty-three percent of the patients received continuous hospice home care during the last week of life, and 34% died in an inpatient hospice bed. Patients with hospice stays of less than 7 days had a lower likelihood of receiving continuous hospice home care than those who had stays of more than 30 days (adjusted odds ratio (AOR) = 0.81, 95% confidence interval (CI) = 0.75 0.87). Patients with hospice stays of 14 days or less had a greater likelihood of dying in an inpatient hospice bed. Furthermore, patients with stays of less than 7 days who were referred from hospitals were six times likelier to die in an inpatient hospice bed than those who were referred from another source (AOR = 6.40, 95% CI = 5.74-7.14). Patients in nursing homes had a 93% lower likelihood of dying in an inpatient hospice bed than patients in the community without a live-in caregiver (AOR = 0.07, 95% CI = 0.03-0.19). Strong independent associations were observed between several other covariates and the study outcomes, particularly the covariates of which state hospice care was provided in and level of pain intensity. CONCLUSION: Findings suggest that continuous hospice home care in the last week of life is less likely to occur when patients have short hospice stays. Also, the probability of dying in an inpatient hospice bed is substantially greater for patients referred from hospitals and referred closer to time of death. Further work to determine the appropriateness of use of the different levels of hospice care is needed. PMID- 12757567 TI - Partnering managed care and community-based services for frail elders: the care advocate program. AB - OBJECTIVES: To describe a demonstration program that uses master's-level care managers (care advocates) to link Medicare managed care enrollees to home- and community-based services, testing whether referrals to noninsured services can reduce service usage and increase member satisfaction and retention. DESIGN: Using an algorithm designed to target frail, high-cost users of Medicare insured healthcare services, the program partners PacifiCare's Secure Horizons and four of its medical groups with two social service organizations. SETTING: Three care advocates located in two community-based social services agencies using telephone interviews to interact with targeted elders living in the community. PARTICIPANTS: Three hundred ninety PacifiCare members aged 69 to 96 receiving care from four PacifiCare-contracted medical groups. INTERVENTION: The 12-month intervention provides telephone assessment, links to eight types of home- and community-based services, and monthly follow-up contacts. MEASUREMENTS: Sociodemographic characteristics of intervention participants, types of service referrals, and acceptance rates. RESULTS: Lessons learned included the importance of building a shared vision among partners, building on existing relationships between members and providers, and building trust without face-to-face interactions. CONCLUSION: The program builds on current insured case management services and offers a practical bridge to community-based services. PMID- 12757568 TI - Contribution of informant and patient ratings to the accuracy of the mini-mental state examination in predicting probable Alzheimer's disease. AB - OBJECTIVES: To determine whether the accuracy of the Mini-Mental State Examination (MMSE) in predicting future Alzheimer's disease (AD) could be improved by the addition of patient and informant ratings of cognitive difficulties. DESIGN: An inception cohort of nondemented patients followed longitudinally for 2 years. SETTING: Patients referred to a university teaching hospital research investigation by their family physicians because of concerns about memory impairment. PARTICIPANTS: One hundred sixty-five community-residing patients were included who did not have dementia or any identifiable cause for memory impairment. After 2 years, 29 met criteria for AD, and 95 were not demented. MEASUREMENTS: Baseline assessments included MMSE, an Informant Rating Scale, and a Patient Rating Scale of cognitive difficulties. After 2 years, patients were diagnosed following the reference standard for probable AD. Diagnosticians were blind to baseline scores. RESULTS: Age and education were included in all analyses as covariates. The best logistic regression model included the Informant Rating Scale and the MMSE (sensitivity = 83%, specificity = 79%). An empirically reduced six-item model that included two items each from the MMSE, the Patient Rating Scale, and the Informant Rating Scale produced a significantly better model than the one with the full test scores (sensitivity = 90%, specificity = 94%). CONCLUSION: Results indicate that inclusion of informant ratings with the MMSE significantly improved its accuracy in the prediction of probable AD. Replication in a new prospective cohort of nondemented patients is necessary to confirm these findings. PMID- 12757569 TI - Accuracy of collateral source reports in very mild to mild dementia of the Alzheimer type. AB - OBJECTIVES: To examine the reporting accuracy of collateral sources (knowledgeable informants) regarding very mild and mild dementia of the Alzheimer type (DAT) and to identify characteristics associated with collateral source accuracy. DESIGN: Secondary data analysis of initial visits of individuals enrolled in a longitudinal study of healthy aging and Alzheimer disease. SETTING: Urban Alzheimer disease research center. PARTICIPANTS: Pairs of 515 individuals with very mild (n = 203) or mild (n = 312) DAT and their collateral sources. MEASUREMENTS: Collateral sources were asked separately during a semistructured interview by experienced clinicians to report current ability of the individual with DAT in memory, orientation, and judgment and problem solving. The clinical performance of the individuals with DAT in these domains was compared with these predictions. RESULTS: Collateral sources were consistently and significantly accurate in reporting the cognitive capabilities of individuals with very mild and mild DAT. Although all types of collateral sources performed significantly better than chance, individual variables that correlated with collateral source accuracy included spousal relationship; living with the individual with DAT; frequent exposure to the individual; and age, education level, sex and dementia severity of the individual with DAT. CONCLUSION: Collateral sources are accurate in reporting the cognitive capabilities of individuals with DAT, even in the very mild stage of dementia. PMID- 12757570 TI - The association between time since last meal and blood pressure in older adults: the cardiovascular health study. AB - OBJECTIVES: To demonstrate a postprandial hypotensive (PPH) phenomenon in older adults. DESIGN: Observational, prospective cohort study composed of baseline and nine follow-up visits. SETTING: Cardiovascular Health Study, an epidemiological study of risk factors for cardiovascular disease in older adults. PARTICIPANTS: Five thousand eight hundred eighty-eight community-dwelling adults aged 65 and older. MEASUREMENTS: Blood pressure and time since last meal were recorded synchronously at baseline and at follow-up clinic visits. Generalized estimating equations were used to estimate associations between time since last meal and blood pressure and to adjust variance estimates to account for repeated blood pressure measures within subjects across fasting times. RESULTS: Mean systolic and diastolic blood pressures were lower in the first hour after the last meal and were progressively higher through the fourth hour after the last meal than blood pressures measured immediately after the last meal (0 hour: 133.7/68.8 mmHg; 1st hour: 130.1/66.6 mmHg; 4th hour: 136.5/71.1 mmHg). Changes were significant for systolic and diastolic measures (P <.001 for both). Exploratory analyses suggested that the systolic PPH association was more pronounced in women. Little evidence was found that the degree of systolic or diastolic PPH varied by age, race, prevalent cardiovascular disease, heart rate, ejection fraction, treated hypertension or diabetes mellitus, or body mass index. CONCLUSION: These data support previous observations that there is a significant drop in blood pressure within 1 hour after a meal in older adults. Time since last meal may be an important factor to consider when measuring blood pressure in older adults, and perhaps national standards need to be set. PMID- 12757571 TI - Reducing fear of falling in seniors through education and activity programs: a randomized trial. AB - OBJECTIVES: To determine the relative effect of education and activity programs on fear of falling, balance, strength, and health status. DESIGN: A randomized intervention trial with two groups (education and activity); evaluators were blind to group allocation. SETTING: Motor performance laboratory at Queen's University. PARTICIPANTS: Thirty-eight community-dwelling seniors who reported a fear of falling and activity restriction but were free of neurological and mobility-limiting orthopedic conditions. INTERVENTIONS: Programs designed to reduce fear of falling were delivered weekly to groups of three to five seniors for 8 weeks, each session lasting 1 hour. The activity program included low resistance exercises and weight-shifting activities. Education focused on identifying and reducing risk factors for falls. MEASUREMENTS: Balance confidence, activity level, limits of stability (LOS), isokinetic strength, and health status were measured twice preintervention (baseline), postintervention, and 6 weeks later. RESULTS: Both programs reduced fear of falling (P <.006) as ascertained from the balance confidence scores. Differential effects were observed in LOS (P <.05); activity improved balance, whereas education led to modest declines. Gains in perception of health status were limited to physical health for the activity group and mental health for the education group. Benefits were generally sustained at follow-up. CONCLUSION: Improved balance confidence is not intervention-specific, but associated changes in physical ability and health status are a function of the composition of the intervention program. PMID- 12757572 TI - Family involvement in end-of-life hospital care. AB - OBJECTIVES: To examine whether the end-of-life treatment provided to hospitalized patients differed for those who had a family member present at death and those who did not. DESIGN: A retrospective cohort analysis. SETTING: An urban community hospital. PARTICIPANTS: All 370 inpatients who died during a 1-year period. MEASUREMENTS: Medical records were examined for whether life-support treatments were provided or withdrawn, occurrence and timing of do-not-resuscitate (DNR) orders, and use of comfort measures such as narcotics and sedation. RESULTS: DNR orders were written for 85% of patients. For patients who had a DNR order written, the average time from the DNR order to death was 4.8 days. Only 26% of patients had one or more treatments withdrawn. Sixty-seven percent of patients received narcotics before death, and 22% received sedatives. Patients aged 75 and older and African Americans were less likely to have a family member present at death. After adjusting for age and ethnicity, patients who had a family member present at death were more likely to have DNR orders written, to have treatments withdrawn, and to receive narcotics before death. Patients with a family member present at death also had a shorter time to death after DNR orders were written. CONCLUSION: The presence of a family member at death appears to be an indirect measure of family involvement during patients' hospitalization. Family involvement before death may reduce the use of technology and increase the use of comfort care as patients die. PMID- 12757574 TI - A randomized controlled trial of an enhanced balance training program to improve mobility and reduce falls in elderly patients. AB - OBJECTIVES: To evaluate the effectiveness of an enhanced balance training program in improving mobility and well-being of elderly people with balance problems. DESIGN: Prospective, single-blind, randomized, controlled trial. SETTING: District general hospital. PARTICIPANTS: One hundred ninety-nine patients aged 60 and older with a Berg Balance Scale (BBS) score of less than 45. INTERVENTIONS: Six weeks enhanced balance training consisting of a series of repetitive tasks of increasing difficulty specific to functional balance. The control group received physiotherapy conforming to existing practice in elderly patients with mobility problems. MEASUREMENTS: Ten-meter timed walk test (TWT), BBS, Frenchay Activities Index (FAI), Falls Handicap Inventory (FHI), and European Quality of Life questionnaire (Euroqol) measured at 6, 12, and 24 weeks after intervention. RESULTS: The mean age +/- standard deviation of subjects was 82.7 +/- 5.6, and baseline characteristics were comparable between the groups. Both groups showed improvements in TWT (intervention: 22.5-16.5 seconds, P =.001; control: 20.5-15.8 seconds, P =.054), BBS (intervention: 33.3-42.7, P =.001; control: 33.4-42.0, P <.0001), FAI (18-21, P =.02 in both groups), FHI score (intervention: 31-17, P =.0001; control: 33-17, P =.0001) and Euroqol score (intervention: 58-65, P =.04; control: 60-65, P =.07). There were no intergroup differences at any time. More patients reported increased confidence in walking indoors (36% vs 28%; P =.04) and outdoors (27% vs 18%; P =.02) in the enhanced balance-training group. CONCLUSION: Exercise programs significantly improve balance and mobility in patients with balance problems, independent of strategy. Enhanced balance training may, in addition, improve confidence and quality of life but needs further investigation. PMID- 12757573 TI - Is a fall just a fall: correlates of falling in healthy older persons. The Health, Aging and Body Composition Study. AB - OBJECTIVES: To identify factors associated with falling in well-functioning older people. DESIGN: Cross-sectional analyses of report of falls over the past 12 months using baseline data from the Health, Aging and Body Composition Study. SETTING: Clinic examinations in Pittsburgh, Pennsylvania, or Memphis, Tennessee. PARTICIPANTS: Three thousand seventy-five high-functioning black and white elderly aged 70 to 79 living in the community. MEASUREMENTS: Physical function assessed using self-report and performance measures. Health status indicators included diseases, medication use, and body composition measures. RESULTS: Almost one-quarter (24.1%) of women and 18.3% of men reported at least one fall within the year before the baseline examination. Fallers were more likely to be female; white; report more chronic diseases and medications; and have lower leg strength, poorer balance, slower 400-meter walk time, and lower muscle mass. In men, multivariate logistic regression models showed white race (adjusted odds ratio (OR) = 1.4, 95% confidence interval (CI) = 1.2-1.6), slower 6-meter walk speed (OR = 1.1, 95% CI = 1.0-1.3), poor standing balance (OR = 1.2, 95% CI = 1.0-1.4), inability to do 5 chair stands (OR = 1.7, 95% CI = 1.3-1.9), report of urinary incontinence (UI) (OR = 1.5, 95% CI = 1.1-2.0), and mid-quintile of leg muscle strength (OR = 0.6, 95% CI = 0.4-0.9) to be independently associated with report of falling. In women, benzodiazepine use (OR = 1.6, 95% CI = 1.0-2.6), UI (OR = 1.5, 95% CI = 1.2-1.9), and reported difficulty in rising from a chair (OR = 1.4, 95% CI = 1.2-1.6) were associated with past falls. CONCLUSION: Falls history needs to be screened in healthier older adults. Even for well-functioning older persons, specific correlates of falling can be identified to define those at risk. PMID- 12757575 TI - Paradoxical vasodilation during lower body negative pressure in patients with vasodepressor carotid sinus syndrome. AB - OBJECTIVES: To elucidate the pathophysiological mechanism of the vasodepressor form (VD) of carotid sinus syndrome (CSS) by maneuvers designed to induce generalized sympathetic activation after baroreceptor unloading (lower body negative pressure, LBNP) or direct peripheral adrenoreceptor stimulation via local administration of norepinephrine (NA). DESIGN: Subjects were identified with VD of CSS through diagnostic testing. SETTING: Research laboratory. PARTICIPANTS: Eleven young controls (YC) (mean age +/- standard error of mean = 22.8 +/- 0.7), eight elderly controls (EC) (72.6 +/- 0.6), and eight elderly patients with VD (78.7 +/- 1.7). MEASUREMENTS: Forearm arterial blood flow (FABF) was measured in the left and right arms by venous occlusion plethysmography. Measurements were performed during baseline conditions, LBNP (-20 mmHg), and intra-arterial NA infusion in the left brachial artery at three progressively increasing rates (60, 120, and 240 pmol/min). RESULTS: During LBNP, FABF significantly decreased in YC (baseline 3.61 +/- 0.30 vs -20 mmHg 2.96 +/- 0.24 mL/100 g/min, P =.030) and EC (4.05 +/- 0.74 vs 3.69 +/- 0.65 mL/100 g/min, P =.033) but increased in elderly patients with VD (3.65 +/- 0.60 vs 4.54 +/- 0.80 mL/100 g/min, P =.020). During NA infusion, a significant forearm vasoconstriction occurred in YC (FABF left:right ratio 1.00 +/- 0.05 at baseline; 0.81 +/- 0.08 at 60 pmol/min, P =.034; 0.81 +/- 0.05 at 120 pmol/min, P <.001; 0.72 +/- 0.04 at 240 pmol/min, P <.001), whereas no significant FABF changes were observed in EC (1.04 +/- 0.06; 0.96 +/- 0.07, P =.655; 0.89 +/- 0.10, P =.401; 0.94 +/- 0.10, P =.590) or elderly patients with VD (1.04 +/- 0.06; 1.16 +/-0.10, P =.117; 1.04 +/- 0.08, P =.602; 1.11 +/- 0.10, P =.305). CONCLUSION: VD of CSS is associated with a paradoxical vasodilatation during LBNP and an impairment of peripheral alpha-adrenergic responsiveness, which may be age-related. PMID- 12757576 TI - Geriatric medicine curriculum consultations for family practice residency programs: american academy of family physicians residency assistance program/hartford geriatrics initiative. AB - Increasing the quality and quantity of geriatric medicine training for family practice residents is a particular challenge for community-based programs. These programs have an average of only seven full-time equivalent physician faculty. This report summarizes results of the Residency Assistance Program/Hartford Geriatric Initiative (RAP/HGI) geriatric medicine curriculum consultations for family practice (FP) residency programs conducted from 1996 to 2001. This project was developed as part of the RAP in family practice. Ten experienced FP educators were selected and trained as special consultants. Between 1996 and 2001, 39 FP residency programs participated in the 1- to 4-day RAP/HGI consultations. The programs were diverse in size and location. The consultations reached 308 family practice residency faculty members involved in training 807 residents. Program evaluations of the consultants were uniformly in the very good to excellent range, with a mean rating of 4.6 (5-point scale, with 5 indicating excellent). At the end of the initial consultation visit, the residency program faculty and the consultant developed short-term goals for geriatrics program development. Eighty five percent (33/39) of the programs submitted their curriculum goals in writing. The mean number of goals per program was 4.8 (range = 3-11). Of the 33 programs with written goals, follow-up was documented for 29 programs. Seventy-nine percent of the programs' self-defined educational goals were met during the 6 to 12 months of follow-up (range 50-100%). Ten of the programs implemented all of their educational goals. The RAP/HGI project demonstrated that achievable geriatric medicine curriculum improvements could occur as part of an onsite consultation process. PMID- 12757577 TI - Barriers to and benefits of leisure time physical activity in the elderly: differences across cultures. AB - The purpose of this study was to compare ethnic differences in attitudes toward barriers and benefits of leisure-time physical activity (LTPA) in sedentary elderly Mexican (MAs) and European Americans (EAs). An in-home, cross-sectional survey was performed on 210 community-dwelling elders from 10 primary care practices in south Texas that are part of the South Texas Ambulatory Research Network, a practice-based research network. Analytical variables included ethnicity, age, sex, income, education, marital status, and LTPA. Fisher exact test was used to analyze the 100 sedentary elders (LTPA <500 kcal/wk; 63 MAs and 37 EAs). Self-consciousness and lack of self-discipline, interest, company, enjoyment, and knowledge were found to be the predominant barriers to LTPA in both groups. Both groups held similar beliefs about benefits gained from exercise, such as improved self-esteem, mood, shape, and health, but the beliefs about the positive benefits of exercise were more prevalent in MAs. These findings remained after adjusting for age, income, education, marital status, and sex. Some might think that a major barrier lies in misconception about benefits of LTPA, but in this study, both ethnic groups were accurate in their perceived benefits of LTPA. When attempting to engage elderly in LTPA, it is important not only to consider what barriers exist but also what beliefs about the benefits exist. PMID- 12757578 TI - Guidelines abstracted from the American Academy of Neurology's Dementia Guidelines for Early Detection, Diagnosis, and Management of Dementia. PMID- 12757579 TI - The revolution in aging is here. 1953. PMID- 12757580 TI - Still waiting for the revolution in aging. PMID- 12757582 TI - The value of informant reports for assessment and prediction of dementia. PMID- 12757583 TI - AGS guidelines on persistent pain in older persons: lack of specific pharmacotherapeutic recommendations. PMID- 12757584 TI - Neuropsychiatric symptoms in the elderly: let us not forget celiac disease. PMID- 12757585 TI - Lack of association between thyroid disorders and Alzheimer's disease in older persons: a cross-sectional observational study in a geriatric outpatient population. PMID- 12757587 TI - Efficacy of antithrombotic therapy for atrial fibrillation in the oldest old. PMID- 12757586 TI - The effect of stability of oral anticoagulant therapy upon patient-perceived health status and quality of life. PMID- 12757588 TI - Donepezil and athetosis in an elderly patient with Alzheimer's disease. PMID- 12757590 TI - Relationship between body composition and cytokines in cachectic patients with chronic obstructive pulmonary disease. PMID- 12757591 TI - Vitamin D and disability. PMID- 12757592 TI - Plasma brain natriuretic peptide levels increase in proportion to changes in pulmonary artery pressure in old-old persons. PMID- 12757595 TI - Unenhanced helical computed tomography in the evaluation of acute flank pain. AB - OBJECTIVES: The purpose of this study was to determine the value of unenhanced helical computed tomography (UHCT) in the diagnosis of acute flank pain at our institution. METHODS: Two hundred and thirty-three consecutive UHCT examinations, performed for suspected renal/ureteral colic between July 2000 and August 2001 were reviewed, along with pertinent medical records. RESULTS: Ureteral calculi were identified in 148 (64%) examinations, evidence of recent passage of calculi was found in 10 (4%) and no calculi were found in 75 (32%). Thirty-two of the conservatively managed patients were excluded for inadequate follow-up. In the remaining 201 patients, sensitivity of UHCT in diagnosing calculi was 99% and specificity was 98%, while the positive predictive value was 99% and negative positive predictive value was 98%. Overall, an alternative or additional diagnosis was established in 28 (12%) patients. Upon diagnosis of ureterolithiasis on UHCT, none of the patients required additional imaging studies for confirmation. CONCLUSION: UHCT is a highly sensitive imaging modality for the detection of urinary tract calculi and obstruction. PMID- 12757596 TI - Association of absence of intestinal oxalate degrading bacteria with urinary calcium oxalate stone formation. AB - AIM: Urinary concentration of oxalate is considered an important factor in the formation of renal stones. Dietary oxalate is a major contributor to urinary oxalate excretion in most individuals. Furthermore, oxalate degrading bacteria have been isolated from human feces. We investigated the significance of oxalate degrading bacteria for urinary oxalate excretion and urinary stone formation. METHODS: Twenty-two known calcium oxalate stone-forming patients (stone formers) and 34 healthy volunteers (non-stone formers) were included in the study. Stool specimens were inoculated into pepton yeast glucose (PYG) medium supplemented with oxalate under anaerobic condition at 37 C for one week. After the incubation period, each colony was checked for the loss of oxalate from the culture medium. A 24-h urine sample was collected in 43 individuals and analyzed for oxalate excretion. RESULTS: Twenty-eight of 34 (82%) healthy volunteers and 10 of 22 (45%) calcium oxalate stone formers were colonized with oxalate degrading bacteria. Calcium oxalate stone formers were more frequently free of oxalate degrading bacteria (P < 0.01). Urinary excretion of oxalate in those with oxalate degrading bacteria was significantly less than in those without oxalate degrading bacteria (P < 0.05). Hyperoxaluria (> 40 mg/day) was found in four of 27 individuals (15%) with oxalate degrading bacteria compared to seven of 16 (44%) without oxalate degrading bacteria (P < 0.05), suggesting an association between the absence of oxalate degrading bacteria and the presence of hyperoxaluria. CONCLUSION: The absence of oxalate degrading bacteria in the gut could promote the absorption of oxalate, thereby increasing the level of urinary oxalate excretion. The absence of oxalate degrading bacteria from the gut appears to be a risk factor for the presence of absorptive hyperoxaluria and an increased likelihood of urolithiasis. PMID- 12757597 TI - Functional evaluation of one-stage urethroplasty with parameatal foreskin flaps repair of hypospadias using uroflowmetry. AB - BACKGROUND: Uroflowmetry is a simple, accurate and non-invasive test. In the present study, we aimed to determine the role of uroflowmetry in the evaluation of the functional results of one-stage urethroplasty with parameatal foreskin flaps (OUPF) technique. METHODS: Twenty-one children who had undergone OUPF repair at our clinic were selected. Selection criteria were that patients were toilet trained and had no fistula. Uroflowmetry was performed using a rotating disk sensor. The maximum flow rate (Qmax) and average flow rate (Qave) were plotted against body surface related flow rate nomograms. The upper 95% tolerance limits for the 5th, 10th, 15th, 20th and 25th percentiles of the normal population were used for comparison. The flow pattern was classified as bell ring, plateau or intermittent. RESULTS: The median age at the first uroflowmetry was 4.7 years (range 2.5-8.6) and the mean postoperative follow-up period was 25 months (range 1-58). Twelve children had Qmax above the 25th, six between the 5th and 25th and three less than the 5th percentiles of the normal population. A normal bell-shaped flow curve was obtained in 17 (80.6%) of the children. Of the three children with Qmax below the 5th percentile, two children had a plateau flow pattern and were found to have a urethal stricture. Dilation was performed successfully, after which the Qmax returned to the normal range and the symptoms disappeared. CONCLUSION: The OUPF technique provided satisfactory functional results for hypospadias repair. We advocate the use of uroflowmetry for routine postoperative follow-up. PMID- 12757598 TI - Transition zone biopsy and prediction of extraprostatic extension at radical prostatectomy. AB - BACKGROUND: There is limited data in the literature that suggests that transition zone (TZ) biopsy might be useful for the prediction of extraprostatic extension (EPE) in clinically localized prostate cancer. We studied the role of TZ biopsy in the prediction of EPE. METHODS: Transition zone biopsies were performed in addition to systematic peripheral zone (PZ) biopsies between November 1995 and December 1999. During this period, 59 patients underwent radical prostatectomy for clinically localized disease. Final pathological results were compared with preoperative clinical and biopsy findings. RESULTS: Of the 59 patients who underwent radical prostatectomy, 46 had cancer only in the PZ cores and 13 had cancer both in the PZ and the TZ cores at the biopsy. Final histopathological results revealed EPE in 19 (32%) patients and positive surgical margins in 22 (37%). In univariate analysis of age, prostate-specific antigen (PSA), mean percentage of positive PZ cores, mean biopsy Gleason score and positive TZ biopsy, there was a significant difference for serum PSA levels (P = 0.021), presence of positive TZ cores (P = 0.018) and percentage of positive PZ cores in patients with and without EPE (P < 0.001). In multivariate analysis, the single independent predictor of EPE was the percentage of positive PZ biopsy cores (P = 0.0227). There was agreement between the side of positive TZ biopsy and EPE in seven of eight patients. CONCLUSION: Taking two TZ cores in addition to peripheral sextant biopsy did not result in better prediction of EPE. The relationship between TZ involvement and the presence of EPE can be investigated further in radical prostatectomy specimens. PMID- 12757602 TI - Significance of capsular attachment and invasion of cancer tissues in prostate cancer. AB - BACKGROUND: There is a discrepancy in tumor node metastasis (TNM) staging of capsular attachment and invasion; the condition was classified as pT3 in 1987, then as pT2 in 1992. Because capsular finding associated with radical prostatectomy is an important prognostic factor, the present study was conducted to characterize clinicopathological states of cancer tissues attached to and invading the capsule. METHODS: Specimens removed by radical prostatectomy exhibiting pT2 or pT3 from 90 patients who did not receive any treatment before surgery were classified as Loc (24%, cancer tissues localized and apart from capsule), Inv (59%, attached to and invading but not penetrating capsule) and Pen (17%, penetrating capsule). Their clinicopathological profiles were examined. RESULTS: Gleason score, volume of cancer tissues, seminal vesicle invasion, positive surgical margin and regional lymph node metastasis of Inv were distributed between those of Loc and Pen. Postoperative management was decided as routine check-up or endocrine therapy according to pathological findings. Median follow-up was 59 months. Prostate-specific antigen (PSA) relapse occurred in 13 patients, one of whom died of prostate cancer. The remaining of these patients lived. Rate of PSA relapse was not different between Loc and Inv, mainly due to endocrine therapy to Inv with high risk factors. CONCLUSION: Pathological profile of Inv lies between those of Loc and Pen. Therefore, pT2a (1997) would be subclassified as Loc and Inv. Patients with Inv may be required to receive the respective management according to clinicopathological profile, which would be different to that of Loc. PMID- 12757603 TI - Prevalence of elevated serum prostate-specific antigen in rural Nigeria. AB - BACKGROUND: Recent hospital and cancer registry data show increasing prostate cancer incidence in Nigeria, which was previously regarded as a low incidence region. This study investigates the prevalence of prostate cancer risk in a previously unscreened cohort of rural Nigerians. METHODS: Rural Nigerian men, 40 years and older, were screened by serum prostate-specific antigen (PSA) and digital rectal examination (DRE) and those with PSA >/= 4 ng/mL and/or abnormal DRE were referred for prostate biopsy. RESULTS: Of 200 consecutive men invited, 151 (75.5%) presented for screening, the mean age was 56.45 + 15.1 and 95 (61.6%) were >/= 50 years of age. Of the 140 who consented to a blood test, PSA correlated with age (r = 0.3, P < 0.01), 14 (10.0%) had abnormal PSA >/= 4 ng/mL, increasing from 3 (3.6%) in men < 60 years to 4 (50%) in men >/= 80 years. The rate was 13 (15.7%) for men >/= 50 years and there was no evidence of increased incidence of prostatitis in the community. Mean (median) PSA in ng/mL increased from 1.17 (0.60) in the youngest to 13.75 (4.45) in the oldest cohort. Of those who accepted DRE, 38 (29.0%) had an enlarged prostate, including two who had nodular prostate, one-third with symptoms, increasing from 4 (5.4%) in those < 50 years to 6 (75.0%) in men >/= 80 years. The proportion of men with PSA >/= 4 ng/mL among those with enlarged vs normal prostate is 27.0 to 3.4%, P < 0.001, and the pattern was similar for men >/= 60 years and those < 60 years of age. The 40 (32.0%) men referred for prostate biopsy defaulted mainly because they did not fully understand the need for further investigation because they were symptom free or afraid of the possible side-effects of the procedure or diagnosis of cancer. CONCLUSION: The proportion of men with PSA >/= 4 ng/mL is comparable to that of previously unscreened populations with high incidence of prostate cancer such as African-American men. A larger study is required to confirm these findings and intensify efforts to determine the prostate cancer detection rate by biopsy in this population. A prostate cancer awareness and education campaign will be useful in this community. PMID- 12757604 TI - Prominent induction of cyclin B1 in G2/M renal cancer cells with butyrolactone 1. AB - BACKGROUND: Butyrolactone 1 (BL) is a cyclin dependent kinase (CDK) inhibitor derived from Aspergillus terreus. None of the present drugs are effective for the treatment of renal cell carcinoma. The use of BL is expected to promote a new type therapy of renal cancer. METHODS: We investigated three human renal cancer cell lines: ACHN, OS-RC-2 and RCC10RGB, using 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) assay and two-color flow cytometry. Simultaneous measurements of DNA content and cyclin expression allowed us to perform cell cycle specific analysis. Western blot analysis was performed using ACHN to represent cell lines. RESULTS: BL inhibited cell proliferation and caused cell accumulation at G2/M phase associated with the emergence of the third peak. Moreover, BL induced cyclin B1 over-expression in G2/M cells. These changes were quite definite, whereas cyclins D1, E and A showed no changes at all. Cyclin B1 accumulation was confirmed by western blot analysis. The chronological observation revealed that the emergence of the third peak preceded the regression of the increased cyclin B1 positive G2/M cells. These results suggested that BL accelerated cyclin B1 accumulation in G2/M cells, which then shifted to G1 phase without cell division. New G1 cells started DNA synthesis most likely as endoreduplication to form the third peak and the mechanism of cyclin B1 accumulation converted into down-regulation. CONCLUSION: BL induced significant cell kinetic interference in the tested human renal carcinoma cell lines. This might indicate the possibility of a new medical treatment modality for renal cancer. PMID- 12757606 TI - Bladder smooth muscle cell phenotypic changes and implication of expression of contractile proteins (especially caldesmon) in rats after partial outlet obstruction. AB - BACKGROUND: The purpose of the present study was to investigate morphological changes in bladder smooth muscle of rats with partial outlet obstruction. We investigated smooth muscle cell phenotypic changes and implication of synthetic phenotype in contractility decrease and bladder compliance after bladder outlet obstruction. METHODS: Partial bladder outlet obstruction was introduced in female rats. Bladder were removed at 1, 3, 6, 10 and 20 weeks after the obstruction. Temporal pattern of changes in bladder mass, light microscopic pathogenesis and phenotypic expression of the bladder smooth muscle cells in the electron micrographs were investigated. Expression of contractile protein was also investigated by the immunoblotting method. RESULTS: Marked increase in bladder mass with marked thickening of smooth muscle layer was observed at 1 week after obstruction. The ratio of myocytes exhibiting contractile and synthetic phenotypes was almost constant until 6 weeks after the obstruction, but thereafter, synthetic phenotypes gradually increased and the ratio (synthetic/contractile phenotype) was 1.5-fold at 20 weeks after the obstruction. Caldesmon was most markedly expressed after the obstruction among contractile proteins examined by the immunoblotting method. CONCLUSION: Phenotypic changes were confirmed in bladder smooth muscle, and the decrease of the ratio of contractile phenotype was observed after long-term obstruction of the bladder outlet. Among the contractile proteins in the bladder smooth muscle cell, caldesmon was considered a reliable marker for predicting the pathogenetic conditions of the bladder. PMID- 12757605 TI - Cadmium nephrotoxicity and evacuation from the body in a rat modeled subchronic intoxication. AB - BACKGROUND: Cadmium (Cd) is an important industrial pollutant, although its mechanism of toxicity has not been completely clarified. We studied Cd-induced subchronic nephrotoxicity and the cadmium evacuation system in rats and cultured human renal tubular cells. METHODS: Male Sprague-Dawley rats were subcutaneously injected with 0.6 mg Cd/kg per day for periods of 3, 5 and 8 weeks. The concentration of Cd in urine, serum and kidneys was measured by atomic absorption spectrophotometry. Nephrotoxicity was evaluated based on the urinary concentration of beta2 microglobulin (B2MG) and histopathological findings. Apoptotic cells were detected by nick-end labeling and DNA laddering, and were based on the level of caspase-3 activity. Cadmium-induced toxicity was also studied in cultured human renal tubular cells. RESULTS: Nephrotoxicity was detected after 4 weeks of exposure to Cd, because Cd and B2MG appeared in urine. The tissue concentration of Cd increased linearly throughout the 8 weeks of exposure to Cd. The concentration of renal Cd did not change in the 3-week exposure group, but it decreased after withdrawal of Cd in the 5-week exposure group, suggesting an active Cd excretion mechanism started after the 4th week. The threshold Cd concentration for nephrotoxicity was 150 micrograms/gram wet tissue, at which concentration histological tubular damage started. Although the kidneys presented mainly necrosis, apoptosis was observed at weeks 4 and 5, before renal tubular necrosis occurred. In vitro DNA laddering was observed and peak caspase-3 activity was detected when the cells were exposed to the threshold concentration of Cd. CONCLUSION: Cadmium was effectively evacuated from the body by exfoliation of damaged renal tubular cells presenting focal tubular necrosis after the renal Cd concentration reached the threshold. Apoptosis may be involved in the regulation of Cd-induced nephrotoxicity. PMID- 12757607 TI - Acute urethritis caused by Neisseria meningitidis. AB - A 48-year-old heterosexual Japanese man visited the outpatient clinic of Nagoya Urology Hospital, complaining of burning pain at voiding and pus discharge from the urethral orifice. These symptoms appeared the day following oral-genital contact (fellatio) with a commercial sex worker. On the basis of the presumptive clinical diagnosis of gonorrhea because of the microscopic detection of diplococci in the urethral discharge, he was treated with levofloxacin (300 mg per day) for 7 days. His symptoms responded quickly and urinalysis taken 7 days later was normal. Microbiological examinations isolated Neisseria meningitidis in the urethral discharge by culture with the use of enzymatic profiles. Further prevalence of sexually transmitted diseases (STD) through oral-genital contact would lead to an increase in meningococcal urethritis. PMID- 12757608 TI - Clear cell adenocarcinoma of the male urethra. AB - We present a rare case of clear cell adenocarcinoma of the male urethra. These tumors are usually presented with hematuria, obstructive voiding symptoms or urinary retention. Histologically, they display tubulocystic, tubular, papillary or diffuse patterns with clear and hobnail cells. The present case of this rare disease emphasizes the aggressive nature of urethral clear cell adenocarcinoma in males. PMID- 12757609 TI - Testicular metastasis from squamous cell carcinoma of the lung. AB - We present a case of squamous cell carcinoma of the testis that metastasized from lung cancer. The patient, who had received left pneumonectomy 2 years earlier for squamous cell carcinoma (SCC) of the lung, developed pulmonary metastasis, which was treated with chemotherapy. Although the recurrence regressed after treatment, the testicular tumor progressed gradually. Left radical orchiectomy was performed. Pathological examination revealed metastatic SCC. Testicular metastasis from lung cancer is a very rare disease. PMID- 12757610 TI - Japan's share of research output in urology and nephrology. AB - BACKGROUND: Recent studies have documented Japan's share of articles in different biomedical fields. However, Japan's contribution to urology and nephrology is not known. METHODS: Seven journals in this field with the highest impact factors were selected to obtain relevant data. In June 2002, we searched the Medline database to obtain country affiliation details of articles published in the selected journals during 1991-2000. The proportion of articles with a higher grade of evidence (randomized controlled trials or clinical trials, cohort studies and case-control studies) were also generated and compared to the average values for all articles published in these journals. RESULTS: Of the total number of articles (22 232), Japan's share of research output for the selected journals in urology and nephrology was 6% (1335 articles) and it ranked second in the world following the USA (56.5%). The recent increase in the share was statistically significant for Japan, Austria, France, Germany, Spain and Turkey. The proportion of articles with a high grade of evidence was lower for articles originated in Japan than for the total number of articles. CONCLUSION: Japan's contribution to urology and nephrology research is within the range of its contribution to other biomedical fields. However, the number of high quality clinical research conducted in Japan in this field was small. PMID- 12757611 TI - Experience with flexible video cystoscope equipped with built-in high-frequency cauterizing element for transurethral resection of the bladder. AB - We successfully treated three patients using the flexible video cystoscope (CYF 240A, Olympus, Japan) for superficial bladder cancer under local anesthesia and/or spinal anesthesia. The major advantage of this new technique is that surgeons can use digital pictures and high frequency cauterization. This new system provides good quality of life for bladder cancer patients experiencing frequent recurrences. PMID- 12757613 TI - Analysis of differential immune responses induced by innate and adaptive immunity following transplantation. AB - The roles of innate and adaptive immunity in allograft rejection remain incompletely understood. Previous studies analysing lymphocyte deficient or syngeneic graft recipients have identified subsets of inflammatory chemokines and cytokines induced by antigen independent mechanisms. In the current study, we analysed a panel of 60 inflammatory parameters including serum cytokines, intragraft chemokines and cytokines, receptors, and cellular markers. Our results confirmed the up-regulation of a subset of markers by innate mechanisms and also identified a subset of parameters up-regulated only in the context of an adaptive response. Thus, we successfully differentiated markers of the innate and adaptive phases of rejection. Current paradigms emphasize that innate signals can promote a subsequent adaptive response. Interestingly, in our studies, expression of the markers induced by innate mechanisms was markedly amplified in the allogeneic, but not syngeneic or lymphocyte deficient, recipients. These results suggest that inflammatory mediators can have functional overlap between the innate and adaptive responses, and that the adaptive component of the rejection process amplifies the innate response by positive feedback regulation. PMID- 12757614 TI - Rat bone marrow-derived dendritic cells, but not ex vivo dendritic cells, secrete nitric oxide and can inhibit T-cell proliferation. AB - The relationships between different dendritic cell (DC) populations are not clearly established. In particular, it is not known how DC generated in vitro relate to those identified in vivo. Here we have characterized rat bone marrow derived DC (BMDC) and compared them with DC isolated from spleen (SDC) and pseudo afferent lymph (LDC). BMDC express typical DC markers and are mostly OX41 positive and CD4 negative. In contrast to ex vivo DC, some BMDC express Fc receptors. FcR+ and FcR- BMDC express similar levels of major histocompatibility complex class II molecules (MHC) and are B7 positive, but some FcR- BMDC express high levels of B7. In contrast to freshly isolated or cultured ex vivo SDC and LDC, both BMDC subpopulations can express inducible nitric oxide synthase (iNOS) and can secrete nitric oxide (NO) in amounts similar to those secreted by peritoneal macrophages. Despite expressing MHC class II and B7, FcR+ BMDC stimulate only a very weak MLR and inhibit stimulation by FcR- BMDC and ex vivo DC. Inhibition is only partially NO dependent. FcR+ BMDC are not macrophages, as judged by adherence and phagocytosis. Both subpopulations are able to present antigen to primed T cells in vitro and are able to prime naive CD4 T cells in vivo. However, unlike SDC, BMDC are unable to stimulate cytotoxic T-lymphocyte (CTL) responses to a minor histocompatibility antigen. Thus, BMDC show marked differences to ex vivo DC and their relationship to those of in vivo DC populations, to date, is unclear. PMID- 12757612 TI - Innate self recognition by an invariant, rearranged T-cell receptor and its immune consequences. AB - This review attempts to illuminate the glycolipid antigen presentation properties of CD1d, how CD1d controls the function of natural T (iNKT) cells and how CD1d and iNKT cells interact to jump-start the immune system. It is postulated that the CD1d-iNKT cell system functions as a sensor, sensing alterations in cellular lipid content by virtue of its affinity for such ligands. The presentation of a neo-self glycolipid, presumably by infectious assault of antigen-presenting cells, activates iNKT cells, which promptly release pro-inflammatory and anti inflammatory cytokines and jump-start the immune system. PMID- 12757616 TI - Age-related increase of peripheral CD4+ CD8+ double-positive T lymphocytes in cynomolgus monkeys: longitudinal study in relation to thymic involution. AB - The age-related increase of peripheral CD4+ CD8+ double-positive (DP) T cells in cynomolgus monkeys has been reported previously. Because the percentage of DP T cells in cynomolgus monkeys increases abruptly in parallel with the thymic involution occurring at around 11 years of age, it was suggested that thymic involution was associated with this increase. Therefore, a longitudinal study was carried out over 5 years to clarify the exact time when DP T lymphocytes start to increase in relation to the thymic involution. Twelve cynomolgus monkeys at 6 years of age were classified into three groups, based on their percentage of DP T cells, as follows: DP-High (>5% DP T cells); DP-Middle (1-5% DP T cells); and DP Low (<1% DP T cells). In the DP-High group, the percentage of DP T cells showed an abrupt increase, of >10%, in monkeys at 7 years of age, and the prevalence of this subset correlated with a distinctive increase in the percentage of memory T cells (CD4+ CD29(high), CD8+ CD28-), indicating an association with the maturation of immune function, including thymic involution. To assess the thymic function, the coding joint of T-cell receptor excision circles (cjTREC) levels in sorted T cells were analysed by polymerase chain reaction (PCR)-enzyme-linked immunosorbent assay (ELISA). The cjTREC in the T cells of the DP-High group (4362 +/- 3139 copies/105 T cells) was significantly lower than that (22 722 +/- 4928 copies/105 T cells) of the DP-Low group. Moreover, the mean copy number of cjTREC in naive T cells was also significantly different between the DP-High and the DP Low group (0.457 +/- 0.181 and 1.141 +/- 0.107, respectively). These findings suggest that thymic involution has an influence on the age-related increase of DP T cells in cynomolgus monkeys. PMID- 12757615 TI - Isolation of proliferation factor of immature T-cell clone in concanavalin A stimulated splenocyte culture supernatant. AB - An athymic mouse-derived immature T-cell clone, N-9F, was not maintained by interleukin-2 alone but required another soluble factor, contained in concanavalin A-stimulated rat splenocyte culture supernatant, namely T cell growth factor (TCGF), for its proliferation. An N-9F-proliferation factor (NPF) was isolated in a pure form from TCGF. N-9F cells and immature thymocytes proliferated in the presence of NPF at 10-11-10-8 g/ml in a dose-dependent manner, but adult thymocytes were not stimulated by NPF. NPF increased DNA synthesis of N-9F. NPF increased CD4 and CD8 double negative thymocytes and CD8 single positive thymocytes in fetal thymus organ culture. A hamster anti-NPF antiserum possessing the capacity to neutralize N-9F proliferation activity of NPF decreased double negative thymocytes. The amino-terminal amino acid sequence of NPF was identified to be Ser-Leu-Pro-Cys-Asp-Ile-Cys-Lys-Thr-Val-Val-Thr-Glu Ala-Cys-Asn-Leu-Leu-Lys-Asp- and was identical to that of rat saposin A. The apparent molecular weight of NPF, 16000, was comparable to that of saposin A. A rabbit anti-mouse recombinant His-tag (mrH)-saposin A antibody recognized a 16000 MW molecule in TCGF. A Hitrap-saposin A antibody column bound NPF and pulled down the NPF activity in TCGF. Thus, NPF in TCGF was a saposin A-like protein possessing the capacity for growth and differentiation of immature thymocytes. PMID- 12757618 TI - NIM-R7, a novel marker for resting B1 and marginal-zone B lymphocytes, is also expressed on activated T and B cells. AB - In mice, follicular B cells have been studied in detail, while two other B-cell subpopulations - marginal-zone B and B1 cells - are less well understood. In this work we report the expression pattern of p58, a lymphocyte-activation marker, recognized by rat monoclonal antibody, NIM-R7, and present on the latter two cell subpopulations. Staining with NIM-R7 showed that undisturbed marginal-zone B cells, as well as peritoneal cavity and splenic B1a cells, constitutively expressed p58, whereas follicular B cells and resting T lymphocytes did not. Ontogenic analysis of different compartments showed that p58 did not appear at any stage of development, prior to the development of mature T or B2 lymphocytes. Upon polyclonal stimulation, however, p58 appeared on both T and B2 lymphocytes. Finally, ricin A-conjugated NIM-R7 was able to kill the BCL1 lymphoma without effect on mature resting B2 cells. Therefore, p58 may be a potential target for diagnosis or therapy of B1 and marginal-zone B-cell malignancies. PMID- 12757617 TI - Costimulation through OX40 is crucial for induction of an alloreactive human T cell response. AB - The alloreactive immune response is a series of events initiated by the interaction of T cells with allogeneic dendritic cells (DCs), involving alloantigen recognition and costimulatory signals. In this study, we investigated the role of OX40 in alloreactivity in vitro. We first demonstrated that anti-OX40 ligand (anti-OX40L) monoclonal antibody (mAb) could markedly suppress the mixed leucocyte reaction (MLR) of peripheral blood mononuclear cells (PBMC). To further define the contribution of the OX40/OX40L system to the MLR, we set up a co culture system of CD4+ T cells and allogeneic monocyte-derived dendritic cells (DCs). After 2 days, OX40 expression was induced on CD4+ T cells and this induction was strongly inhibited by the addition of cytotoxic T lymphocyte associated antigen-4 (CTLA-4)-Fc fusion protein, suggesting that the expression of OX40 during alloreaction is dependent on CD28 signalling. Next we examined the effects of anti-OX40L mAb, CTLA-4-Fc fusion protein and anti-human leucocyte antigen (HLA)-DR mAb on the proliferative response of CD4+ T cells to allogeneic DCs. The proliferation of T cells was almost completely suppressed by anti-OX40L mAb, which was comparable with that of CTLA-4-Fc. Measurement of interleukin-2 (IL-2) production in the culture supernatants showed that suppression of a proliferative response was at least in part ascribed to reduced IL-2 production. Furthermore, purified OX40L- allogeneic DCs could induce considerable proliferation of CD4+ T cells, which was suppressed by anti-OX40L mAb. These results suggest that the OX40/OX40L system is crucial for induction of the allogeneic T-cell response and the OX40/OX40L system is subsequent to and dependent on CD28 signalling, but is crucial for the end outcome of the human alloreactive T-cell response. PMID- 12757619 TI - Hyporesponsiveness of murine B lymphocytes exposed to the filarial nematode secreted product ES-62 in vivo. AB - ES-62 is a phosphorylcholine (PC)-containing glycoprotein secreted by filarial nematodes, parasites of vertebrates including humans. We have previously demonstrated that pre-exposure to this molecule in vitro interferes with subsequent B-cell receptor (BCR)-dependent activation of murine splenic B lymphocytes. To investigate the significance of this during filarial nematode infection, we now employ mice exposed to ES-62, at concentrations equivalent to those found for PC-containing molecules in the bloodstream of parasitized humans, via release from implanted osmotic pumps. Using this approach, we reveal that splenic and lymph node mononuclear cells, and also purified splenic B cells recovered from these mice have reduced ability ex vivo to proliferate in response to BCR ligation. The effect on BCR-induced proliferation was further investigated with respect to elucidating the mechanism of action of the parasite product and was shown to be associated with impaired signal transduction affecting the ErkMAPkinase pathway. Also, it was found that ES-62 did not act by promoting apoptosis or by priming for apoptosis following subsequent stimulation, but rather, appeared to render cells hyporesponsive to stimulation. ES-62 is thus shown for the first time to be a potent modulator of B lymphocyte function in vivo at a concentration relevant to natural filarial nematode infection. This finding considerably strengthens the idea that ES-62 plays a role in evasion of the immune response during parasitism. PMID- 12757620 TI - Soluble FcgammaRIIa inhibits rheumatoid factor binding to immune complexes. AB - Soluble low-affinity receptors for IgG are known to inhibit immune complex (IC) mediated inflammation, and expression by leukocytes is elevated in several inflammatory diseases. Immunoglobulin M (IgM) rheumatoid factors (RF), anti-Fc autoantibodies, are found in autoimmune diseases, such as rheumatoid arthritis (RA), as well as in normal immune responses. This study demonstrated that soluble FcgammaRIIa inhibits the interaction of rheumatoid factors with ICs. The recombinant soluble low-affinity FcgammaR, rsFcgammaRIIa, partially inhibited (30 70%) the rate of precipitation of soluble ICs by RF-positive RA sera. This required the normal interaction of FcgammaRIIa with Fc as the effect could be abrogated with the Fab fragment of the blocking mAb IV-3. Furthermore, rsFcgammaRIIa partially inhibited (40%) the binding of a monoclonal IgM RF (RF AN) to an IC formed by IgG2 antibody binding to an antigen-coated biosensor chip. Since RF-AN has been characterized by crystallography to bind to the CH2/CH3 interface of the IgG-Fc, and leukocyte FcgammaRIIa binds to a distinct site centred on the lower hinge, this inhibition is uncompetitive. Some inhibition (15%) of staphylococcal protein A binding to IC was also observed. As soluble FcgammaRIIa disrupts Fc:Fc interactions in IgG-ICs, we propose that this alteration of the IC also reduces the accessibility of Fc portions in the IC, resulting in the partial inhibition of ligands, particularly IgM RF, which bind Fc. We propose that the high concentrations of soluble FcgammaR found during inflammation can affect the properties of ICs and their interaction with the immune system. PMID- 12757621 TI - Effect of cytofectins on the immune response of murine macrophages to mammalian DNA. AB - DNA, depending on base sequence, can induce a wide range of immune responses. While bacterial DNA is stimulatory, mammalian DNA is inactive alone and can, moreover, inhibit the response to bacterial DNA. To determine whether the mode of cell entry affects the immune properties of mammalian DNA, we have investigated the effects of the cytofectin agents Fugene 6 (Roche Diagnostics Corp., Indianapolis, IN), Lipofectin and Lipofectamine (Life Technologies, Grand Island, NY) on the responses of murine macrophages to DNA from calf thymus and human placenta. Whereas calf thymus and human placenta DNA alone failed to stimulate J774 or RAW264.7 cell lines or bone marrow-derived macrophages, these DNAs in complexes with cytofectin agents stimulated macrophages to produce nitric oxide but not interleukin 12. Both single-stranded and double-stranded DNAs were active in the presence of cytofectins. Macrophage activation by the DNA-cytofectin complexes was reduced by chloroquine, suggesting a role of endosomal acidification in activation. As shown by flow cytometry and confocal microscopy, the cytofectins caused an increase in the uptake of DNA into cells. Our findings indicate that macrophages vary in their response to DNA depending on uptake pathway, suggesting that activation by DNA reflects not only sequence but also context or intracellular location. PMID- 12757622 TI - Host's innate immune response to fungal and bacterial agents in vitro: up regulation of interleukin-15 gene expression resulting in enhanced natural killer cell activity. AB - Natural killer (NK) cells play an important role in the first line of defence against viral infections. We have shown earlier that exposure of human peripheral blood mononuclear cells (PBMC) to viruses results in rapid up-regulation of NK cell activity via interleukin-15 (IL-15) induction, and that this mechanism curtails viral infection in vitro. By using Candida albicans, Escherichia coli and Staphylococcus aureus, we now show here that exposure of PBMC to fungi and bacteria also results in an immediate increase of NK cytotoxicity. Reverse transcriptase-polymerase chain reaction and Western blot analyses as well as the use of antibodies against different cytokines revealed that IL-15 induction played a predominant role in this NK activation. These results indicate that IL 15 is also involved in the innate immune response against fungal and bacterial agents. PMID- 12757623 TI - Assessment of oral transmission using cell-free human immunodeficiency virus-1 in mice reconstituted with human peripheral blood leucocyte. AB - Oral-genital contact is one of the risk factors for the transmission of human immunodeficiency virus (HIV) in adults. In recent reports, oral exposure to simian immunodeficiency virus (SIV) was found to have important implications for the achievement of mucosal transmission of HIV in a rhesus monkey animal model. In the present study, we aimed first to establish a small animal model which did not develop tonsils suitable for HIV oral mucosa transmission, using non-obese diabetic/severe combined immunodeficiency (NOD/SCID) mice and NOD/SCID B2m(null) mice grafted with human peripheral blood leucocytes (hu-PBL) and stimulated with interleukin (IL)-4, and second to investigate whether oral exposure to cell-free R5 and X4 HIV-1 could lead to oral transmission of HIV through intact or traumatized mucosal tissues in humanized mice. Both low and high concentrations of cell-free R5 and X4 viruses failed to cause oral transmission with or without trauma in hu-PBL-NOD/SCID and NOD/SCID Beta2m(null) mice, which presented a number of CD4+ cells in gingival tissues and oral cavities with or without tissue injury. The present results show that IL-4-administrated NOD/SCID B2m(null) mice are useful as a small-humanized model for the study of HIV oral infection, which may help to define the window of opportunity for oral transmission by the HIV virus in animal model experiments. PMID- 12757624 TI - Infection with chicken anaemia virus impairs the generation of pathogen-specific cytotoxic T lymphocytes. AB - Infection with chicken anaemia virus (CAV), a circovirus, can result in immunosuppression and subsequent increased susceptibility to secondary infections. This is the first report of impairment of pathogen-specific cytotoxic T lymphocytes (CTL) after natural and experimental infection of chickens with CAV and Marek's disease virus (MDV) or reticuloendotheliosis virus (REV). MDV- and REV-specific CTL were generated at 7 days post infection by 9-30-day-old-chickens that were positive for maternal antibodies to CAV at 9-17 days of age. Replication of CAV could not be demonstrated in these chickens using quantitative real-time polymerase chain reaction (PCR) and reverse transcriptase (RT)-PCR assays. In contrast, REV-specific CTL failed to develop when chickens negative for maternal antibodies at 9-17 days of age were infected. Infection with CAV at 45 days of age after CAV maternal antibodies had waned also caused a decreased REV-specific CTL response. In these chickens increased levels of CAV DNA of up to 107 copy numbers per micro g DNA and increased relative transcript levels of CAV by up to a factor of 106 were detected by quantitative real-time PCR and RT-PCR. Interleukin (IL)-1beta and IL-2 mRNA levels were not significantly affected by CAV infection at 7 or 14 days p.i. Similar assays for interferon-gamma (IFN gamma) transcripts demonstrated a 10-fold increase in IFN-gamma mRNA levels at 7 days post infection following REV or REV + CAV infection, while CAV alone caused a two- to fourfold increase. These results show a strong link between CAV antibody status, CAV replication, and the ability to generate REV-specific CTL. It is likely that the immunosuppressive effects of subclinical infection have previously been underestimated. PMID- 12757625 TI - Increased interleukin-10 expression is not responsible for failure of T helper 1 immunity to resolve airborne Mycobacterium tuberculosis infection in mice. AB - With a view to determining whether failure of mice to resolve Mycobacterium tuberculosis (Mtb) infection is a consequence of downregulation of T helper 1 (Th1) immunity by interleukin (IL)-10, mice deleted of the gene for IL-10 were compared with wild-type (WT) mice in terms of their ability to make IL-10 mRNA, generate Th1-mediated immunity [as measured by synthesis of mRNA for interferon gamma (IFN-gamma)], IL-12p40 and inducible nitric oxide synthase (iNOS), and to control lung infection. It was found that the response of WT mice to infection included a substantial and sustained increase in IL-10 mRNA synthesis in the lungs. A Th1 response in the lungs of WT and IL-10-/- mice was evidenced by a large and sustained increase in the synthesis of mRNA for IFN-gamma, IL-12p40 and iNOS, with somewhat higher levels of these mRNA species being made in the lungs of IL-10-/- mice, particularly at an early stage of infection. However, IL-10-/- mice were no more capable than WT mice at combating infection. PMID- 12757626 TI - Immunotherapy of spontaneous mammary carcinoma with fusions of dendritic cells and mucin 1-positive carcinoma cells. AB - The tumour-associated antigen mucin 1 (MUC1) is a multifunctional protein involved in protection of mucous membranes, signal transduction, and modulation of the immune system. More than 70% of cancers overexpress MUC1, making MUC1 a potential target for immunotherapy. In the present study, MUC1 transgenic mice were crossed with syngeneic strains that express the polyomavirus middle-T oncogene (PyMT) driven by the mouse mammary tumour virus promoter long-terminal repeat (MMTV-LTR). The resultant breed (MMT mice) developed spontaneous MUC1 expressing mammary carcinomas with 100% penetrance at 8-15 weeks of age. As found in human breast cancer, the mammary carcinoma in MMT mice arose in multiple stages. Immunization with fusions of dendritic cells and MUC1-positive tumour cells (FC/MUC1) induced MUC1-specific immune responses that blocked or delayed the development of spontaneous breast carcinomas. In contrast, there was no delay of tumour development in MMT mice immunized with irradiated MC38/MUC1 tumour cells. The efficacy of fusion cells was closely correlated with the timing of initial immunization. Immunization with FC/MUC1 initiated in MMT mice at < 1, 1-2 and 2-3 months of age rendered 33, 5 and 0% of mice free of tumour, respectively, up to 6 months. Whereas mice immunized in the later stage of tumour development succumbed to their disease, immunization resulted in control of tumour progression and prolongation of life. These results indicate that immunization with FC/MUC1 can generate an anti-MUC1 response that is sufficient to delay the development of spontaneous mammary carcinomas and control tumour progression in MMT mice. PMID- 12757627 TI - The combination of plasmid interleukin-12 with a single DNA vaccine is more effective than Mycobacterium bovis (bacille Calmette-Guerin) in protecting against systemic Mycobacterim avium infection. AB - Sub-unit vaccines utilizing purified mycobacterial proteins or DNA vaccines induce partial protection against mycobacterial infections. For example, immunization with DNA vaccines expressing the gene for the immunodominant 35000 MW protein, common to Mycobacterium avium and Mycobacterium leprae but absent from the Mycobacterium tuberculosis complex, conferred significant protection against infection with either virulent M. avium or M. leprae in mice. However, the level of protection was equivalent to that obtained with the viable, attenuated vaccine, Mycobacterium bovis, bacille Calmette-Guerin (BCG). The cytokine, interleukin (IL)-12, is essential for priming naive CD4+ T lymphocytes to differentiate into interferon-gamma (IFN-gamma)-secreting T cells. We have used a novel self-splicing vector expressing both chains of murine IL-12 to determine if plasmid IL-12 would increase the efficacy of a vaccine expressing the M. avium 35000 MW protein (DNA-Av35). Co-immunization with p2AIL-12 and DNA Av35 led to a significant increase in the number of antigen-specific IFN-gamma secreting cells and total amount of IFN-gamma released, but a concomitant fall in the antibody response to the 35000 MW protein. This pattern of response was associated with enhanced clearance of M. avium from the liver and spleen of coimmunized mice, and was significantly more effective than BCG or DNA-Av35. alone. Following M. avium challenge there was significant increase in the expansion of the 35000 MW antigen-reactive T cells in the coimmunized mice. Therefore, plasmid-delivered IL-12 acts as an effective adjuvant to increase the protective efficacy of a single DNA vaccine against M. avium infection above that achieved by BCG, and this strategy may improve the efficacy of subunit vaccines against M. leprae and M. tuberculosis. PMID- 12757631 TI - Regional comparisons of inpatient and outpatient patterns of cerebrovascular disease diagnosis in the province of Alberta. AB - The diagnosis of cerebrovascular disease (CBVD) from administrative data has been critically examined by epidemiologists in recent years. Much of the existing literature suggests that hospital discharge diagnoses based on ICD-9-CM codes are an unreliable source of information for determining a diagnosis of stroke, particularly when four- and five-digit codes are used. We examined how diagnoses for CBVD in hospital inpatient and outpatient facilities vary between rural and urban areas and among the 16 administrative health regions. Our analysis revealed differences in diagnostic patterns between the two sources of data, differences between rural and urban areas, and variation across most of the regions. Geographic variation in health service utilization, diagnostic practices, specialty of the physician making the diagnosis, and disease burden may explain our findings. Our results suggest that the diagnosis of patients attending rural facilities are either coded differently (and less precisely) than those of urban residents or are coded more precisely only after the patients attend urban facilities. Regional differences in coding practices show that any CBVD surveillance system based on administrative data requires a large-scale (in this case, province-wide) and person-oriented approach. PMID- 12757630 TI - Agreement between proxy- and case-reported information obtained using the self- administered Ontario Familial Colon Cancer Registry epidemiologic questionnaire. AB - Case-control studies of fatal cancers often rely on proxy respondents. Therefore, it is important to determine the completeness and accuracy of proxy-reported information. We evaluated proxy reports using the Ontario Familial Colon Cancer Registry epidemiology questionnaire. A proxy questionnaire was completed by spouses or relatives identified by a sample of participating cases. Item non response and percentage agreement (between case and proxy reports) were assessed. More than 30% of proxies were unable to report on physical activity, gynecological surgery, alcohol intake, weight 20 years ago, and oral contraceptive use. Proxy reports of medical history and bowel screening varied, the percentage missing ranging from 5% for diabetes to 44% for familial polyposis in the case of medical history, and from 4% for colonoscopy to 27% for hemoccult tests in the case of screening. Agreement between case and proxy report was good to excellent for colonic screening, most medical history, and for reproductive, medication and vitamin use variables (74% to 100%). It is useful to collect proxy information on such variables as medical history, parity, colonic screening and vitamin use, whereas oral contraceptive use and previous weight are not well reported. PMID- 12757632 TI - Collection and retention of demographic, medical, and occupational information in northeastern Ontario workplaces. AB - This study determined whether workplaces in northeastern Ontario currently collect and retain demographic information, medical history, work history, and information on occupational exposure. Surveys were mailed to 434 northeastern Ontario workplaces with 50 or more employees, and a telephone follow-up was conducted. The response rate was 42.6% (185/434). Over 97% of workplaces reported that they always collect surnames and first names of employees, 13.5% reported collecting maiden names (and 70.8% never collect maiden names), 85.4% collect date of birth, 55.7% next of kin, 97.8% current address, 21.6% medical history, and 31.9% collect the health insurance number. Job titles were routinely recorded by 79.5%. Start and end dates for each job were always recorded by 68.1%, and 70.3% reported that they always note the area of work. Overall, 64.9% of workplaces collected previous place of employment. For 72.1%, legislation influenced the amount of information collected on current records. Thirteen percent routinely recorded smoking history on occupational health records, and 25.9% undertook exposure surveillance. This type of information can assist in planning occupational epidemiological studies. PMID- 12757633 TI - The Ontario sun safety working group. PMID- 12757634 TI - A call for action to support best practices in evaluation of comprehensive tobacco control evaluation strategies. AB - The National Tobacco Control Best Practices Working Group convened a two-day workshop to support best practices in evaluation of comprehensive tobacco control strategies. A Better Practices Model, aimed at developing a self-correcting system for best practices, guided the workshop content and process. Organizers intended to identify a common surveillance and monitoring framework for tobacco control strategies in Canada by first building strong working relationships between 44 decision-makers, practitioners and researchers from 12 Canadian jurisdictions. Participants identified needs and recommendations related to increased understanding and use of uniform evaluation strategies, building capacity, and recognition of the complexity of the task of evaluating comprehensive tobacco control strategies. The workshop highlighted the need for increased communication to facilitate understanding across the different sectors of participants. It also identified the potential benefits of harmonization in evaluation of tobacco control strategies across jurisdictions. Priority actions include forming a national team to agree on a model for evaluation, conducting an environmental scan for indicators, planning evaluation / monitoring and research agendas and determining roles for various stakeholders. PMID- 12757635 TI - [Microsurgery for intracranial aneurysm, clinical analysis of 1,041 cases]. AB - OBJECTIVE: To study the effect of microsurgery on incranial aneurysm and factors influential in prognosis. METHODS: A retrospective analysis was made on the clinical data of 1,041 patients, 505 males and 536 females, aged 44.2 +/- 12.6, with incranial aneurysm treated by microsurgery 1988 approximately 2001, including clipping of aneurysm (982 cases, 94.3%), isolating of the aneurysm (32 cases, 3.1%), and coating of the aneurysm (27 cases, 2.6%). RESULTS: 93 patients (8.9%) had multiple aneurysms. 357 aneurysms (31.2%) were in the posterior communicating artery, 261 (22.8%) in the anterior communicating artery, 210 (18.4%) in the internal carotid artery, 158 (13.8%) in the middle cerebral artery, 63 (5.5%) in the anterior cerebral artery, 59 (5.2%) in the vertebral basilar artery, and 35 (3.1%) in the posterior cerebral artery. The overall operative mortality was 1.8%. Angiography performed post-operationally showed that the aneurysm was occluded in 978 patients (99.6%). The prognosis was related to the age of patient (P = 0.001), Hunt and Hess grading before operation (P = 0.001), diameter of aneurysm (P = 0.000), and whether the aneurysm ruptures during operation (P = 0.006), and was not related to the gender of patient (P = 0.059). CONCLUSION: Clipping of the aneurysm is considered the best treatment. Microinvasive neurosurgery has improved significantly the effect of treatment of intracranial aneurysm. Factors influencing prognosis after operation include the age of patient, Hunt and Hess grading before operation, diameter of aneurysm, and whether the aneurysm ruptures during operation. PMID- 12757636 TI - [Combined intraarterial thrombolysis and intra-cerebral stent for acute ischemic stroke institute of brain vascular diseases]. AB - OBJECTIVE: To test feasibility, safety and efficacy of combined intraarterial thrombolysis and intra-cerebral stent for acute ischemic stroke. METHODS: From 2001-5 to 2002-8, 19 patients with acute onset of stroke were treated with intraaterial thrombolysis followed by intra-cerebral stent. 9 located in middle cerebral artery (MCA), 10 located in basal artery (BA). For each patient, intraarterial thrombolysis of MCA or BA was applied at first, stent angioplasty was applied at the stenosis left after the thrombolysis by using the stent for coronary artery. RESULTS: All the 19 patients left stenosis after intraaterial thrombolysis, and the average stenosis was 85% in diameter. After the operation of stent, the images showed the vascular is smooth and had no stenosis left. The symptoms were disappeared or improved. Thrombus formation in the stent occurred on 1 patient 24 hours after the operation. TIA never occurred in the left 18 patients then. TCD follow up showed blood flow is normal. CONCLUSIONS: It is feasibility of intraaterial thrombosis followed by intra-cerebral stent. And it is valid for preventing occlusion again of the vessel and decreasing the rate of TIA occurring. PMID- 12757637 TI - [Anatomical study and three dimensional image characteristic analysis of basicranial artery and its clinical significance]. AB - OBJECTIVE: To observe and measure the inside diameter of basicranial arteries, the angulation of main arteries, the three dimensional image characteristic of internal carotid arteries and the anatomical variation of Willis circle. METHODS: The arteries of 30 formalin-fixed adult heads were injected with latex after which the caliber and characteristic of cerebral arteries were observed and measured. The three dimensional image characteristic of internal carotid arteries and its branches were measured using 3D-DSA. RESULTS: (1) Main artery caliber: origin of internal carotid artery (Left 5.12 +/- 1.48 mm; Right 5.11 +/- 1.42 mm); origin of middle cerebral artery (Left 2.93 +/- 1.44 mm; Right 2.92 +/- 1.46 mm); origin of anterior cerebral artery (Left 2.63 +/- 1.33 mm; Right 2.61 +/- 1.32 mm); origin of vertebral artery (Left 4.37 +/- 1.21 mm; Right 3.22 +/- 1.64 mm); origin of basilar artery (4.45 +/- 1.28 mm); origin of posterior cerebral artery (Left 2.62 +/- 1.36 mm; Right 2.61 +/- 1.22 mm). (2) The angulation of main arteries: C1, 2 of ICA and C4, 5 of ICA (Left 32 +/- 22 degrees; Right 36 +/ 28 degrees ); ICA and ACA (Left 43 +/- 26 degrees; Right 46 +/- 28 degrees). (3) The results show that anatomical and three dimensional image characteristic of internal carotid arteries have no difference (P > 0.05). (4) The anatomical variation of Willis circle: Type O (56.7%); Type A (16.7%); Type P (20.0%); Type AP (6.7%). CONCLUSIONS: It is helpful to measure the inside diameter of basicranial arteries for the selection of various catheter in interventional neuroradiology, to observe the angulation of main arteries and the three dimensional image characteristic of internal carotid arteries for the moulding of various catheter in endovascular therapy and to master the anatomical variation of Willis circle for decreasing complications of endovascular treatment and judging prognosis of cerebrovascular diseases. PMID- 12757639 TI - [Multifetal pregnancy reduction of triplets to twins: a prospective comparison of pregnancy outcome]. AB - OBJECTIVE: To investigate the outcome of triplet pregnancy and the necessity and benefit of multifetal pregnancy reduction (MFPR). METHODS: Sixty cases of triplet pregnancy were collected. In 15 cases that did not choose MFPR, the triplets were spontaneously obtained. Out of the 45 cases with triplets obtained after infertility treatment, 32 chose reduction to obtain twins using transvaginal puncture of the fetal heart and physical hurt. The maternal obstetric complication rate, preterm delivery rate, means gestational age at delivery, birth weight of infants and percentage of fetal weights < 1,500 g were compared between these two groups. RESULTS: The maternal obstetric complication rate and preterm delivery rate were lower significantly in the reduction group. Deliveries occurred earlier slightly in the triplets group compared with the reduction group. The mean birth weight of infants was higher and the percentage of low weight infants (< 1,500 g) was lower in the reduction group. CONCLUSION: MFPR of triplets to twins is effective to improve pregnancy outcome. MFPR applied on triplets is reasonable and should be accepted if requested. PMID- 12757638 TI - [Endovascular stent-assisted angioplasty for atherosclerotic intracranial stenosis, a clinical analysis of 39 cases]. AB - OBJECTIVE: To assess the preliminary outcome of endovascular stent-assisted angioplasty for intracranial stenosis. METHODS: Thirty-nine patients with atherosclerotic intracranial stenosis (19 cases in posterior circulation and 20 in anterior circulation) were treated with endovascular angioplasty using balloon expandable coronary stents. Follow-up was made for 4 - 24 months. The clinical manifestations were observed and CT, MR, or DSA were conducted before and after the operation. RESULTS: All of the stents were successfully implanted on the first procedure without procedural and periprocedural complications. The mean carotid artery stenosis rate was 73.5 +/- 6.7% before the procedure and 11.2 +/- 4.3% after the procedure. The patients were clinical asymptomatic and neurologically intact. Follow-up angiographies made in 29 patients revealed no restenosis. CONCLUSION: With effective short-term results and helpful in decreasing incidence of stroke, endovascular stenting is safe and feasible for treatment of intracranial stenosis. PMID- 12757641 TI - [Surgical treatment of newborns with congenital heart diseases]. AB - OBJECTIVE: To summarize the experience of surgical treatment of newborns with congenital heart diseases. METHODS: The experience of surgical treatment of 8 newborns with critical congenital heart diseases, including 3 cases of D transposition of great arteries with intact ventricular septum, 2 cases of ventricular septal defect with artrial defect (ASD), one case of complete atrioventricular canal defect, 1 case of obstructed supracardiac total anomalous pulmonary venous drainage with ASD, and 1 case of patent ductus arteriosus (PDA). RESULTS: The case of PDA underwent ligation under normothermic anesthesia and the other 7 cases were operated upon under moderate or deep hypothermic cardiopulmonary bypass. All the 8 cases were observed in ICU for 2 approximately 10 days, and were discharged 7 approximately 15 days after operation. The follow up after discharge showed a satisfying outcome. The postoperative complications included low cardiac output, infection of mediastinum, and respiratory distress syndrome. CONCLUSION: The critical and complex congenital heart diseases should by diagnosed as early as possible and emergency operation is effective and feasible. PMID- 12757640 TI - [A novel molecular mechanism of congenital FV deficiency: mutation in the intron acceptor splice site of human blood coagulation FV gene]. AB - OBJECTIVE: To explore the molecular mechanism involved in patient with congenital FV deficiency. METHODS: Activity of FV was determined by biochemical method. The PCR products of FV gene was analyzed by DNA sequencing directly or cloned into T vector prior to DNA analysis. The mutation of FV gene in proband and his family numbers was analysed by restriction enzyme analysis. Its occurrence was investigated in the control group. DNA was extracted from the peripheral blood mono1nuclear cells of the proband, male, 18 years old, and his parents. The PCR products were analyzed by direct sequencing or cloned into T-vector prior to DNA analysis. One hundred patients with different kind of hemotopathy were used as controls. RESULTS: A single point mutation, AG-->GG was found at position 3' splice site of intron 8 of the proband. This mutation was confirmed by family screening. CONCLUSION: A single point mutation, AG-->GG at position 3' splice site of intron 8 mutation of FV gene is related to the pathogenesis of congenital FV deficiency. PMID- 12757642 TI - [Inhibiting scar formation in rat cutaneous wounds by blocking TGF-beta signaling]. AB - OBJECTIVE: TGF-beta plays a key role in wound scarring. This study explored the possibility of using gene therapy to inhibit wound scarring by blocking TGF-beta signaling. METHODS: In vitro, human normal dermal fibroblasts were infected with recombinant adenoviruses of truncated TGF-beta receptor II (tTGF-betaRII, 100 pfu/cell) and beta-galactosidase (beta-gal, 100 pfu/cell), and their effects on regulating TGF-beta1 gene expression were analyzed by Northern blot. For gene therapy, beta-gal and tTGF-betaRII viruses (1 x 10(9) pfu)were injected intradermally at left and right side of dorsal skin of newborn Sprague-Dawley rats (n = 15) respectively. A full-thickness incisional wound (0.5 cm long) was created at the injection sites of each rat 2 days post-injection. An incisional wound was similarly created in the middle part of the dorsal skin of tTGF-betaRII transgenic mice (n = 5) and control mice (n = 5). Wound tissues of rats and mice were harvested at various time points post-wounding for histological and immunohistochemical analysis. Scar area in tissue section was measured by Image Pro Plus software. RESULTS: Over-expression of tTGF-betaRII markedly reduced TGF beta1 gene expression in dermal fibroblasts. Adenovirus mediated gene expression in skin reached a peak level 2 - 3 days post-injection, and decreased gradually at 5 - 7 days. Two weeks post-wounding, histology and quantitative analysis demonstrated that relative scar area in the wounds of transgenic mice and control mice were 136,969.8 +/- 66,339 and 474,641.6 +/- 227,396 respectively, the scar area of transgenic wounds was 29 percent of control area (P < 0.05). In all rats, wounds transfected with tTGF-betaRII gene healed with much less scarring (relative scar area 128,311.2 +/- 36,764.6) than control wounds (251,189.1 +/- 62,544.7) of the same rat, with a 45% reduction of scar area in average (P < 0.001). In addition, the tTGF-betaRII expression also decreased inflammation and TGF-beta1 production in treated wounds, and promoted the repair of panniculus muscle in treated wounds. CONCLUSIONS: Adenovirus mediated over-expression of tTGF-betaRII can block TGF-beta signaling and inhibit wound scarring, and thus can serve as a gene therapy strategy to control wound scarring. PMID- 12757643 TI - [Impact of cyclin-dependent kinase inhibitor p27 on resistance of ovarian cancer multicellular spheroids to taxol]. AB - OBJECTIVE: To examine the expression of the cyclin-dependent kinase inhibitor p27 in ovarian cancer multicellular spheroid (MCS) and explore the reversal effect of p27-antisense oligodeoxynucleotide (p27-ASON) on taxol resistance in ovarian cancer cell lines. METHODS: Three-dimensional culture was used to form MCS of human ovarian cancer cell lines A2780 and CAOV3. The MCSs were divided into 3 groups: MCSs transduced with p27-ASON or p27-sense oligodeoxynucleotide (p27-SON) by LipofectAMINE respectively and MCS without transduction. Monolayer A2780 and CAOV3 cells were cultured as controls. The cells were exposed to taxol of different concentrations (0.2, 2.0, 10.0, and 20.0 micro mol/L) for 24 hours. The expression of p27 in those cells was detected with Western blot and flow cytometry (FACS), and the subcellular distribution of p27 was detected by laser confocal microscopy before and after the experiment. The cell cycle profile and apoptosis were analyzed by FACS. The resistance to taxol was detected with trypan blue exclusion testing. RESULTS: Compared with that in monolayer cells, expression of p27 in MCS cells was significantly higher (P(A2780) = 0.011, P(CAOV3) = 0.024). The percentage of cells in G0-G1 phase was significantly higher in the MCS cells than in the monolayer cells, and the percentages of cells in S and G2-M phases were significantly lower in MCS cells than in monolayer cells. The apoptotic rate of the monolayer cells was significantly higher than those of MCS cells treated with taxol of the concentration of 20.0 micro mol/L (P(A2780) = 0.003, P(CAOV3) = 0.015). The apoptotic rate of p27-ASON/MCS was significantly higher than that of MCS (P(A2780) = 0.022, P(CAOV3) = 0.036). There was no significant difference of apoptotic rate between p27-SON/MCS and MCS (P(A2780) = 0.412, P(CAOV3) = 0.071). The monolayer A2780 and CAOV3 cells formed compact spheroids after three-dimensional culture. Cells transduced with p27-SON formed compact MCS too. However, cells tranduced with p27-ASON formed loose collections of cells easy to shatter. The expression of p27 in p27-ASON/MCSs was downregulated. CONCLUSION: The drug resistance of ovarian cancer MCS is related to upregulation of p27. P27-ASON reverses the resistance of ovarian cancer to taxol, thus increasing the chemotherapeutic sensitivity of ovarian cancer cells. PMID- 12757644 TI - [Chronic Helicobacter pylori infection induces apoptosis-resistance in gastric epithelial cells]. AB - OBJECTIVE: To establish a cell model by chronically infecting human gastric epithelial cells with H.pylori, and to determine the effect of chronic H.pylori infection on apoptosis of gastric epithelial cells. METHODS: Human non-tumor gastric epithelial cells GES-1 were cocultured with an H.pylori strain SS1 for 16 weeks, in order to obtain GES-1 "model" cells. Biological characteristics of the model cells including growth, adherence and clone formation were determined. Apoptosis of the "model" cells in response to apoptosis inducers such as H.pylori and other enterobacteria and agents commonly used in the chemotherapy of gastric cancer were measured by flow cytometry. RESULTS: A cell model of human gastric epithelial cells with chronic H.pylori infection (GES-1 model cells) was successfully established. Apoptosis was dramatically decreased in the "model" cells with and without stimulation by H.pylori and other enterobacteria and some chemotherapeutic agents. CONCLUSION: Chronic H.pylori infection induces apoptosis resistance in gastric epithelial cells, which may increase the risk of the development of gastric cancer. PMID- 12757645 TI - [Effect of morphine on expression of gene of enzymes related to purine nucleotide metabolism in c6 glioma]. AB - OBJECTIVE: To investigate the effect of morphine on catabolism and anabolism of purine nucleotide in c6 glioma cells. METHODS: C6 glioma cells were cultured and divided into 3 groups: 1) morphine group: morphine (10 micro g/ml culture) was added for 6 h, 12 h, 24 h, 48 h, and 72 h; 2) morphine + naloxone group: naloxone (1 micro mol/L) was added for 1 hour and then morphine (10 micro g/ml) was added for 24 hours; and 3) control group: normal saline was used for 6, 12, 24, 48, and 72 hours. The C6 glioma cells were centrifuged. RT-PCR was used to examine the gene transcripts of key enzymes of purine salvage way, hypoxanthine-guanine phosphoribosyl transferase (HGPRT) and adenylate kinase (AK). RT-PCR-Southern blotting was used to examine the gene transcripts of key enzymes of purine salvage way, xanthine dehydrogenase (XD)/xanthine oxidase (XO) mRNA. RESULTS: Compared with that in the control group, the transcript of AK mRNA was significantly lower in the C6 cells treated with morphine for 24 hours, and began to re-increase 48 hours after morphine treatment. The transcript of AK mRNA remained at a low level after treatment of naloxone for 1 hour and treatment of morphine for 24 hours. The levels of transcript of HGPRT mRNA were lower in the morphine group than in the control group at all time points after treatment. However, the level of transcript of HGPRT mRNA 72 hours after treatment was higher in the morphine group than in the control group. The level of transcript of HGPRT mRNA 24 hours after exposure to morphine in the naloxon2 + morphine group was still lower than in the control group. The levels of transcripts of XD/XO mRNA were significantly higher after exposure to morphine in comparison with those in the control group at all time points after treatment. However, the levels of XD/XO mRNA 24 hours after exposure to morphine in the naloxone + morphine group recovered to the normal level. CONCLUSION: The downregulation effect of morphine on the gene expression of AK and HGPRT may not be mediated by mu receptor. The upregulation effect of morphine on the gene expression of XD/XO may be mediated by mu receptor. Naloxone reverses the effect of morphine on enhancement of XD/XO gene expression and cannot reverse the inhibitory effect of morphine on HGPRT and AK gene expression. PMID- 12757646 TI - [Subcutaneous transplantation of microencapsulated Chinese hamster ovary (CHO) cells/pcDNA 3.1/mIL-12 and subcutaneous transplantation of microencapsulated CHO/pcDNA3.1/mIL-12 combined with 5-fluoro-uracil in treatment of tumor-burdened mice]. AB - OBJECTIVE: To investigate the effect of subcutaneous transplantation of microencapsulated Chinese hamster ovary cells (CHO)/pcDNA3.1/mIL-12 and subcutaneous transplantation of microencapsulated CHO/pcDNA3.1/mIL-12 combined with 5-fluoro-uracilum (5-FU) in treatment of tumor-burdened mice. METHODS: CHO/pcDNA3.1/mIL-12 and CHO/pcDNA3.1 were suspended in solution of sodium alginate and made into microcapsules. Sixty mice were divided into 6 groups of 10 mice: (1) IL-12 group, microencapsulated CHO/pcDNA3.1/mIL-12 was injected subcutaneously, (2) combined treatment group: CHO/pcDN with 5-FU, (3) 5-FU group: 5-FU was injected intraperitoneally, (4) blank vector group: CHO/pcDNA3.1 was injected subcutaneously, (5) tumor-burdened group: without any treatment, and (6) blank control group: normal mice without any treatment. Except the mice of the blank control group, all mice were injected subcutaneously into the inner side of right thigh with mice colonic adenoma cells. The volume of tumor was measured every third day. 20 days after the treatment, 5 mice in each group were killed to examine the serum Th1 type cytokines: interferon (IFN)-gamma, IL-12, and Th2 type cytokins: IL-4, IL-10 by double antibody sandwich ELISA. The spleens were made into suspension of lymphocytes to examine the activity of natural killer cell (NK) and cytolytic T lymphocyte (CTL). The survival period of the remaining 5 mice in each group was observed till the 60th day. RESULTS: The activity of NK and CTL were significantly much more in the IL-12 group than in other groups. The activity of NK was significantly much more in combined treatment group than in the tumor-burdened group. The levels of Th1 type cytokines were the lowest in the tumor-burdened group. There was no difference in the levels of Th1 type cytokine between the tumor-burdened group and 5-FU group. However, the levels of Th1 type cytokine were significantly higher in the IL-12 group than in other groups. The levels of Th2 type cytokines were rather high in the tumor-burdened group. There was no difference in the levels of Th2 type cytokine between the tumor-burdened group and 5-FU group. However, the levels of Th2 type cytokine were the lowest in the IL-12 group than in other groups. There was no significant difference in volume of tumor among the tumor-burdened group, blank vector group, and 5-FU group. The mean diameters of tumor in the IL-12 group and combined treatment group were significantly smaller than in the 5-FU, tumor-burdened, and blank vector groups (P < 0.05), however, without a difference between the IL-12 group and combined treatment group. The survival periods of the IL-12 group and combined treatment group were significantly longer than those in the blank vector, tumor-burdened and 5-FU groups (P < 0.05). CONCLUSION: Microencapsulated CHO/pcDNA3.1/mIL-12 transplanted subcutaneously significantly improves the immune function of tumor-burdened mice and partially overcomes immune suppression caused by chemotherapy, and is effective in slowing the growth of tumor and lengthening the survival period of tumor-burdened ice. PMID- 12757647 TI - [Effects of oxytocin, misoprostol and nimodipine on expression of L-type voltage dependent calcium channel mRNA of the uterine myometrium and left ventricular myocardial cells in late pregnant rats]. AB - OBJECTIVE: To study the effect of oxytocin, misoprostol and nimodiping on expressions of L-type voltage dependent calcium channel (VDCC-L) alpha(1) and VDCC-L beta(2) mRNA in the myometrium and left ventricular myocardial cells of the late pregnant rats. METHODS: A reverse transcription polymerase chain reaction (RT-PCR) method was used to examine and quantitate VDCC-L alpha(1) and VDCC-L beta(2) mRNA expressions in the uterine myometrium and left ventricular myocardial cells of rats, which were divided into the nature labor group, oxytocin group (oxytocin inducing labor), misoprostol group (misoprostol inducing labor) and nimodipine group (active labor at 48 hours after administering nimodipine). RESULTS: (1) In the uterine myometrium, the levels of VDCC-L alpha(1) mRNA expression in the nature labor, oxytocin, misoprostol and nimodipine group were 0.800 3 +/- 0.165 9, 0.863 1 +/- 0.192 1, 0.812 0 +/- 0.173 4 and 0.742 6 +/- 0.182 6, respectively. No significant difference was found in the four groups (P > 0.05). The levels of VDCC-L beta(2) mRNA expression in the above four groups were 0.646 9 +/- 0.130 4, 0.506 2 +/- 0.147 2, 0.500 5 +/- 0.135 6 and 0.492 9 +/- 0.127 6, respectively. There was no remarkable difference between the nature labor group and the other three administering drugs groups (P > 0.05). (2) In the left ventricular myocardial cells, expressions of VDCC-L alpha(1) in the nature labor, oxytocin, misoprostol and nimodipine group were 0.662 5 +/- 0.180 1, 0.636 5 +/- 0.157 8, 0.591 7 +/- 0.141 3 and 0.542 6 +/- 0.143 6, respectively; the levels of VDCC-L beta(2) mRNA expression were 0.670 2 +/- 0.140 5, 0.606 2 +/- 0.143 9, 0.591 4 +/- 0.121 9 and 0.585 2 +/- 0.131 0, respectively. Although both the levels of VDCC-L alpha(1) and VDCC-L beta(2) mRNA expression in the nature labor group were a little higher than those in the other three experiment groups, the statistic difference was not noted (P > 0.05). CONCLUSIONS: Oxytocin, misoprostol or nimodipine can induce or inhibit labor through regulating expressions of VDCC-L alpha(1) and VDCC-L beta(2) mRNA in the rat uterine myometrium and it may not have an adverse effect on heart function of normal pregnant rats. VDCC-L may be the common channel of labor induced by internal or external factors. PMID- 12757648 TI - [Relationship between p53, bax and bcl-2 expression and cell apoptosis in intrahepatic cholestasis of pregnancy]. AB - OBJECTIVE: To investigate the changes of gene p53, bax and bcl-2 expression and to study the role on cell apoptosis in placenta through apoptosis in placenta with intrahepatic cholestasis of pregnancy (ICP). METHODS: TdT-mediated dUTP nick end labeling (TUNEL) and immuno-histochemistry methods were used for 31 placental samples with ICP and 31 normal placental samples to detect apoptosis index and the expression of p53, bax and bcl-2 in placenta tissues. RESULTS: The apoptosis index in placental cytotrophoblast, syncytiotrophoblast, decidual cells and mediate cell on ICP group was significantly higher than that in control group (P < 0.01). The expression of p53 was higher than those of control. Bax expression in syncytoitrophoblast, decidual cell and mediate cell in ICP group was higher (P < 0.01), but the expression of bax in placental cytotrophoblast cell was not significantly higher (P > 0.05). The expression of bcl-2 of these positions in ICP group was lower than that in control group (P < 0.01) p53, bax and bcl-2 expressions in (chi +/- s)% in the syncytoitrophoblast cells of placenta with ICP is 75.9 +/- 8.2, 65.8 +/- 13.8, 45.3 +/- 11.3. CONCLUSION: The placental dysfunction may closely relate to the apoptosis. Abnormal overexpression of p53, bax and the low expression of bcl-2 on placental tissue of ICP was the main reason of placental apoptosis. PMID- 12757649 TI - [Study on prenatal diagnosis using fluorescence quantitative polymerase chain reaction for congenital toxoplasmosis]. AB - OBJECTIVE: To investigate prenatal diagnosis and treatment of toxoplasmosis in fetuses with fluorescence quantitative polymerase chain reaction (FQ-PCR) technique. METHODS: Of the 70 pregnant women with toxoplasma (TOX) DNA positive, TOX DNA in amniotic fluid and/or fetal umbilical cord blood was detected with FQ PCR technique to diagnose fetal infection. 48 ones were given routine treatment with spiramycin for 2 therapy periods. Ultrasound examination were undertaken in all of pregnant women to monitor fetal growth. RESULTS: Of the 70 cases with TOX DNA positive, TOX DNA was detected in 21 fetuses. TOX DNA positive rates were similar in amniotic fluid and umbilical cord blood. The higher the TOX DNA, the higher fetal infectious rate. Fetal infectious rate was lower in treatment group (21%) than that in control group (50%), there was a statistically difference between two groups. CONCLUSIONS: Maternal TOX infection may cause fetal damage. Detection of TOX-DNA in amniotic fluid with FQ-PCR technique can diagnose fetal toxoplasmosis exactly. Treatment in pregnant period may decrease intrauterine infection rate. PMID- 12757650 TI - [Fetal development and gestational weeks for triplets pregnancies and the comparison with singletons]. AB - OBJECTIVE: To study the fetal developmental regulation and gestational weeks at labor for triplets pregnancies. METHODS: To detect biparietal diameters and femoral length at different gestational weeks, record gestational weeks of labor and birth weights, and compare with those of singletons. RESULTS: The fetal development of triplets slowed down from the beginning of the 28th gestational week compared with that of singletons (the mean difference was 2.1 mm and 3.1 mm respectively, P < 0.01, P < 0.01). Birth weights of 36.2% triplets was less than that of the 10th percent of the same singletons gestational week. 95.7% triplets delivered their babies before 36 weeks. The average gestational week was 34.0 +/- 1.6, obviously ahead of the singletons'. CONCLUSION: The fetal developmental regulation of triplets pregnancies is different from that of singletons, the gestational period is shorter than those of twins and singletons. PMID- 12757651 TI - [Concentrations of cortisol and dehydroepiandrosterone sulfate in fetal umbilical cord blood for term labor]. AB - OBJECTIVE: To study the effect of cortisol and dehydroepiandrosterone sulfate (DHEA-S) in fetal umbilical cord blood on term labor. METHODS: Cortisol and DHEA S concentrations were measured By radioimmunoassay in 100 term fetal umbilical cord blood. They were divided into four groups. Group A selective cesarean section without any birth pain (n = 18), Group B cesarean section in latent phase (n = 10), Group C cesarean section in active phase (n = 12), Group D spontaneous vaginal deliver (n = 60). RESULTS: The concentrations of fetal umbilical cord cortisol in spontaneous vaginal deliver group was gradually increased with gestational week. The peak level was in the 39th gestation week, by the 42nd gestation week, the concentration of cortisol declined to the 37th gestation week. DHEA-S changed paralleled with cortisol (r = 0.46, P < 0.05). Active birth pain was associated with increased fetal umbilical cortisol concentration, but not with DHEA-S. The concentrations of fetal umbilical cord cortisol in vaginal deliver group was higher than all the cesarean section groups. CONCLUSION: Concentrations of cortisol and DHEA-S played an important role in the initiation and acceleration of labor. PMID- 12757652 TI - [Clinical features of 3798 perinatals suffering from syndromic neural tube defects]. AB - OBJECTIVE: To investigate the patterns and associated malformations in neural tube defects (NTDs) cases. METHODS: From 1987 to 1995, hospital-based cluster sampling method was adopted for collecting data. During that period all live or still births with 28 weeks of gestation or more were assessed within 7 days after delivery. RESULTS: Three thousand seven hundred and ninety eight syndromic NTDs were identified, among which anencephaly, spina bifida and encephalocele were 997, 2394 and 407 respectively. 51.3% associated abnormalities of NTDs appeared in muscle and skeletal system, 19.6% in face, ear and neck, and 9.3% in urinary genital system. Congenital talipes equinovarus, cleft lip with cleft palate, gastroschisis, talipes valgus, undescended testicle were frequently combined with NTDs. The most frequent association in schisis type of NTDs was NTDs accompanied by cleft lip with or without cleft palate (67.5%), followed by anencephaly with cleft palate (8.3%) and anencephaly with omphalocele (6.6%). Of all syndromic NTDs, the rate of low birth weight was 36.9%, of perinatal mortality was 71.2% and the ratio 33.1% patients were diagnosed prenatally. CONCLUSIONS: Approximately one third of NTDs may be associated with other system's malformation. Syndromic NTDs has a high perinatal mortality rate and a poor prognosis. PMID- 12757653 TI - [Analysis of X chromosome mosaicism in patients with premature ovarian failure by fluorescent in-situ hybridization]. AB - OBJECTIVE: To investigate whether low-level 45, X/46, XX mosaicism presents in women with premature ovarian failure (POF) and the sensitivity and specificity of fluorescent in-situ hybridization (FISH) for detecting X chromosome mosaicism. METHODS: Karyotypes of peripheral lymphocyte in 18 patients with POF and 9 normal controls were analyzed and the orange signals during FISH using X chromosome enumeration probes (CEPX) were counted. RESULTS: Single signals of X chromosome (45, X) were found in 7.6% of the total counted cells, which was significantly greater than that in the controls, in spite of the normal 46, XX karyotypes by routine analyses in all POF patients. CONCLUSIONS: This study indicated that some POF patients may attribute to low-level 45, X/46, XX mosaicism. FISH is more sensitive than the routine chromosome analyses in detecting low-level X chromosome monosomy. PMID- 12757654 TI - [Combined effect of interleukin-6 and estrogen receptor gene polymorphisms on bone mass in postmenopausal women]. AB - OBJECTIVE: To examine the individual and combined effects of interleukin-6 (IL-6) and estrogen receptor (ER) gene polymorphisms on the Z score of bone mineral density (BMD). METHODS: BMD at lumbar spine (L(2 - 4)) and femoral neck (FN) were analyzed in 205 postmenopausal Chinese women by dual energy X-ray absorptiometry. Polymorphic 3'flanking region of IL-6 gene and the allelic variance in ER gene PvuII and XbaI sites were studied by polymerase chain reaction. Serum alkaline phosphatase, osteocalcin and urinary pyridinolin were also measured. RESULTS: There were D/D, D/E, C/D, C/C and E/E 5 genotypes of IL-6 gene in our samples, 193 of 205 women were D/D or D/E types. No difference of BMD at L(2 - 4) or FN was found between D/D and D/E types. For the PvuII and XbaI polymorphisms in these IL-6 D/D and D/E genotypes, the FN BMD was higher in pp in comparison with Pp (P = 0.036), and lumbar spine BMD was higher in XX type, comparing with Xx and xx genotype (P = 0.005 and 0.031, respectively). Women without Px haplotype had elevated BMD at L(2 - 4) (P = 0.029) and a trend for higher BMD at FN (P = 0.056) than those with it. Combining the women with IL-6 D/D, D/E genotypes and ER gene Px haplotypes, the L(2 - 4) BMD in those of DD * without Px haplotype showed a tendency to be higher than those of DD * with Px haplotype (P = 0.057), and was significantly increased than those of DE * with Px haplotypes (P = 0.036). CONCLUSION: The introduction of ER gene Px haplotype in the analysis of IL-6 gene might identify individuals with a reduced bone mass more precisely. PMID- 12757655 TI - [Identification of the two subtypes of latent autoimmune diabetes in adults by glutamic acid decarboxylase 65 antibody titers]. AB - OBJECTIVE: To compare the clinical characteristics between type 2 diabetes and latent autoimmune diabetes in adults (LADA) and to define the two distinct types of LADA with different glutamic acid decarboxylase antibody (GADA) titers. METHODS: Sera of 750 patients with an initial diagnosis of type 2 diabetes mellitus (T2DM) were screened for GADA with radioimmunoprecipitation assay. The distribution and frequency of different GADA indices were described. Two hundred and ninety five patients were further studied and divided into four groups (T2DM; GADA index < 0.05; index > or = 0.5 and index > or = 0.05 but < 0.5) to compare the age of onset, body mass index, level of major component of adult hemoglobin (HbA1c) and C peptide as well as the rates of hypertension, hyperlipidemia and chronic complications. RESULTS: A total of 64 antibody-positive patients were identified. Compared with T2DM, these patients had younger age of onset, lower C peptide level (fasting C peptide 500 pmol/L vs 414 pmol/L, P < 0.01), lower body mass index (23.2 kg/m(2) vs 21.2 kg/m(2), P < 0.01) and also lower rates of hypertension (48.7% vs 31.7%, P < 0.05) and hyperlipidemia (60.2% vs 38.5%, P < 0.01). However, only the patients with high GADA titer had reduced beta cell function as compared with T2DM and low titer patients. Their diabetic complications were less than those of T2DM. Low GADA titer (index 0.05 - < 0.5) patients were similar to T2DM patients, except that they were prone to ketoacidemia. CONCLUSION: Two clinically distinct types of LADA can be identified by GADA titers. High titer GADA (GADA > or = 0.5) patients have more resemblance to insulin dependent diabetes and can be regarded as LADA-type 1 diabetes, while low titer GADA patients (0.05 - < 0.5) have clinical and metabolic phenotype of type 2 diabetes and can be regarded as LADA-type 2 diabetes. PMID- 12757656 TI - [Is insulin resistance a common pathway for hereditary and environmental factors induced hypertension?]. AB - OBJECTIVE: To investigate whether insulin resistance is the common route for hereditary and environmental factors to cause hypertension. METHODS: 93 hypertensives with family history of hypertension, 94 hypertensives without family history of hypertension and 99 normal tensives without family history of hypertension as well as their spouse, and one child in each family were enrolled in the present study. Insulin sensitivity was calculated from fasting plasma glucose (FPG) and insulin (FINS) with the formula insulin sensitivity index (ISI) = 1/(FPG x FINS) and insulin resistance index (Homa-IR) = (FPG x FINS)/22.5. The contribution of insulin resistance to blood pressure elevation was investigated by multivariate regression analysis. RESULTS: Subjects with essential hypertension, regardless of the presence or absence of family history of hypertension, were more insulin resistant. Multivariate regression analysis demonstrated that insulin resistance contributed to 17% of mean blood pressure (MBP) elevation, whereas taken FPG + total cholesterol (TC) + high density lipoprotein (HDL) together it explained only 9% of MBP changes in the indicator group. When family history of hypertension was included as a independent variable in the analysis, it became the most important factor instead of insulin resistance in the model and contributed to 30% of the changes of MBP, while contribution of insulin resistance was significantly reduced as to explain only 7% of the changes of MBP. CONCLUSIONS: Insulin resistance is a common route for hereditary and environmental factors to induce hypertension. It is suggested that improving insulin sensitivity may play an important role in the management of essential hypertension. PMID- 12757657 TI - [Immunohistochemical identification of adrenomedullin in adrenal diseases]. AB - OBJECTIVE: Adrenomedullin (ADM) is a hypotensive peptide isolated from human pheochromocytoma tissue. In order to identify the pathophysiological effects of ADM on the etiology of adrenal diseases which can induce hypertension, the localization of immunoreactive adrenomedullin (ir-ADM) in adrenal tissue of subjects with normal adrenal and patients with different adrenal diseases was investigated by means of immunohistochemical staining. METHODS: Peroxidase anti peroxidase complex method was undertaken in the tissue of human adrenal cortex (n = 10) and medulla (n = 10)of subjects with normal adrenal, as well as in the tissue of pheochromocytoma (n = 31), primary aldosteronism including adrenocortical adenoma (n = 13) and hyperplasia (n = 9), Cushing's syndrome including adrenocortical adenoma (n = 12), hyperplasia (n = 10), carcinoma (n = 4) and non-functional adrenocortical carcinoma (n = 7). RESULTS: Positive immunohistochemical staining of ir-ADM was observed in all the cases of normal adrenal medulla and 16 of the 31 cases of pheochromocytoma. The intensity of immunohistochemical staining of ir-ADM in the majority of pheochromocytoma was lower than that in normal adrenal medulla (P < 0.01). The number of pheochromocytoma cases with positive staining in paroxysmal hypertensive group was higher than that in sustained hypertensive group (P < 0.05). Negative immunohistochemical staining was found in all the cases of normal human adrenal cortex, adrenocortical adenoma, hyperplasia and carcinoma. CONCLUSION: It is suggested that heterogeneous of localization of ir-ADM in pheochromocytoma tissue may be correlated to the diversity of blood pressure in patients with pheochromocytoma. The pathophysiological effects of ADM on the etiology of pheochromocytoma should be further investigated. PMID- 12757658 TI - [A case report and pedigree study of a single point mutation in RET proto oncogene and type 2B multiple endocrine neoplasia]. AB - OBJECTIVE: To determine the relationship between a single point mutation in RET proto-oncogene and the occurrence of multiple endocrine neoplasia type 2B (MEN 2B) in a Chinese pedigree. METHODS: We used the methods of polymerase chain reaction (PCR), reverse transcriptase polymerase chain reaction (RT-PCR) and direct gene sequencing of PCR products by an automated DNA sequencer to scan the entire exon 16 of RET proto-oncogene in the tumor (c)DNA from one patient with MEN-2B and the leukocyte DNA from this patient and both of his parents. RESULTS: We found the same mutation Met(ATG)-->Thr(ACG) at codon 918 in exon 16 of RET proto-oncogene in both the tumor (c)DNA and leukocyte DNA of the MEN-2B patient in the form of homozygous missense mutation, but there was no corresponding mutation in leukocytes DNA of his parents. CONCLUSION: We propose that in Chinese population, the point mutation M918T is also associated with the onset of MEN-2B and this case of MEN-2B is sporadic. Thus it may provide a genetic basis for the early diagnosis and treatment of patients suffering from MEN-2B and their at-risk family members in Chinese population. PMID- 12757659 TI - [Changes of serum interleukin (IL)-12 and IL-13 in asthmatic patients and regulatory effects of glucocorticoids on them]. AB - OBJECTIVE: To observe the changes of serum interleukin (IL)-12, IL-13 and lung function in asthmatic patients and to evaluate the influence of glucocorticoid on them. METHODS: The serum samples were obtained from (1) 25 asthmatic patients with acute asthma attack before and after one week course of oral prednisone, (2) 20 asthmatic patients in remission stage, and (3) 15 healthy volunteers. Serum IL 12 and IL-13 were determined with sandwich ELISA. Lung ventilatory function forced expiratory volume in one second (FEV(1)) and respiratory impedance airway resistance (R(5)) were measured in all patients. RESULTS: (1) Serum level of IL 12 in asthma attack group was significantly lower than that in asthma remission group (P < 0.05); both were significantly lower than that in normal control group (P < 0.01); in asthma attack group IL-12 level after prednisone treatment was significantly higher than that before treatment (P < 0.01). (2) Serum level of IL 13 in asthma attack group was significantly higher than that in asthma remission group (P < 0.01); both were significantly higher than that in normal control group (P < 0.01); in asthma attack group IL-13 level after prednisone treatment was significantly lower than that before treatment (P < 0.01). Correlation analysis showed that the serum level of IL-12 was positively correlated to FEV(1) and negatively correlated to R(5) and to serum level of IL-13 (r(1) = 0.458, r(2) = -0.516, and r(3) = -0.549, respectively; P < 0.05 and P < 0.01); the serum level of IL-13 was negatively correlated to FEV(1) and positively correlated to R(5) (r(1) = -0.493, and r(2) = 0.528, respectively; P < 0.05). CONCLUSION: The secretion of IL-12 and IL-13 was impaired in asthma with a significant increase in serum level of IL-13 and decrease in serum level of IL-12. Glucocorticoid could downregulate the serum level of IL-13 and upregulate the serum level of IL 12, redress the imbalance of IL-12/IL-13, and improve lung function in asthmatic patients. PMID- 12757661 TI - [The value of autofluorescence spectrum analysis of gastric juice in diagnosing gastric carcinoma: a preliminary result]. AB - OBJECTIVE: To reveal the difference of autofluorescence spectrums of gastric juice derived from malignancy and benignancy for screening and diagnosis of gastric carcinoma. METHOD: Gastric juice from 202 patients with different gastric diseases were collected, then detected their autofluorescence spectrums (the excitation wavelength is 288 nm, whereas the range of emission wavelength is 300 800 nm) after diluted by 1:10. The diagnostic model for gastric cancer was made by Classification and Regression Trees V2.0 software. RESULTS: There were three peaks (the emission wavelength were 320 - 360 nm, 576 nm and 670 - 690 nm respectively) in the autofluorescence spectrums of all patients, though the intensity of the first peak (with the emission wavelength of 320 - 360 nm) were enhanced distinctively in malignance than those in benignancy. The diagnostic model's sensitivity and specificity of prior probability were 91.4% and 83.2% respectively, whereas the sensitivity and specificity of posterior probability were 85.7% and 82.6%. CONCLUSION: Detection of autofluorescence spectrum of gastric juice has great prospect in the diagnosis and screening gastric carcinoma. PMID- 12757660 TI - [The relationship between subclassification of anti-myeloperoxidase IgG by enzyme linked immuno-sorbent assay analysis and vasculitis activity]. AB - OBJECTIVE: Antineutrophil cytoplasmic antibody (ANCA) is an important serological diagnostic tool for certain vasculitides, such as Wegener's granulomatosis and microscopic polyangiitis. Several studies suggested that ANCA titers correlated with disease activity. However, in some patients, ANCA still remained positive when patients were in clinical remission. The major isotype of ANCA is IgG, which has four subclasses. This study is to investigate the relationship between anti MPO IgG subclasses and vasculitis activity. METHODS: Serum samples, taken from 30 anti-myeloperoxidase (MPO) antibody positive patients both in active phase and in remission, were analysed by ELISA for their anti-MPO IgG subclass distribution and the change of anti-MPO IgG subclasses was compared with the change of anti MPO IgG antibodies. RESULTS: All four anti-MPO IgG subclasses were positive in active phase, IgG(4) being the highest. In remission, the level of anti-MPO IgG(1) and IgG(3) subclasses decreased the most and the percentage of negative change of IgG(3) was the highest, even more significant than that of the total IgG. CONCLUSION: anti-MPO IgG(3) antibody is more closely related with clinical disease activity than the total IgG. Higher ratio of IgG(4) may suggest that chronic repeated antigen stimuli may play a role in the production of ANCA. PMID- 12757662 TI - [An epidemiological and quality of life study of irritable bowel syndrome in Zhejiang province]. AB - OBJECTIVE: To evaluate the prevalence and quality of life (QOL)on irritable bowel syndrome (IBS) patients in Zhejiang province, and to evaluate the applying of Chinese version of the SF-36 questionnaire on IBS. METHODS: From Jan 2001 through Jan 2002, IBS patients from city, island, countryside and suburban in 10 hospitals in Zhejiang province were recruited to the study. 662 Roma II criteria positive IBS patients completed the demographic checklists, symptoms questionnaire and QOL by Chinese version of the SF-36. We compared the demographic characteristics of IBS with the general population in Zhejiang province. The previously reported SF-36 (Chinese vision) data in general population were considered normative QOL. RESULTS: (1) Male to female was 0.89:1. Compared with general population, IBS was more common in female (P < 0.05), adults in 25 - 50 years old (P < 0.001), and among high educated and cadres samples (P < 0.001). (2) The dominant symptoms in IBS included the change of stool formation (79.1%), abnormal of processing of stool (67.9%), abdominal pain (67.7%), abdominal distention (63.2%) and mucous stool (49.8%). Besides colonic symptoms, IBS can cause generalized body discomfort and psychosocial problems. (3) IBS patients experienced significant impairment in QOL. Decrements are most pronounced in vitality, general health, mental health and bodily pain scales. (4) Compared with the general population in Hangzhou city (adjusted to gender and age), IBS patients scored significantly lower on all SF-36 scales (P < 0.001), with the exception of physical function scale. There was a significant correlation between symptom scores and all 8 SF-36 scales. CONCLUSIONS: The prevalence of IBS had a close correlation with sex, age, education and profession. IBS patients experience significant impairment in QOL. The Chinese version of the SF-36 Health Survey Scales may be a useful measurement of IBS patients. These data offer further insight into epidemiology and the impact of IBS on patient functional status and well-being of Chinese. PMID- 12757663 TI - [A study of susceptibility genes mapping for aberrant B1 cell proliferation in New Zealand mice]. AB - OBJECTIVE: To map the susceptibility genes for aberrant B1 cell proliferation in New Zealand mice. METHODS: New Zealand Black (NZB) x New Zealand White (NZW) F1 x NZW backcross mice model was set up and polymorphic microsatellite markers and quantitative trait locus (QTL) analysis was used. RESULTS: Susceptibility genes of aberrant B1 cell proliferation was linked to the microsatellite markers on chromosome 1, 4, 19 in NZB and on chromosome 17 in NZW such as D1Mit115, D4Mit58, D19Mit73 and D17Mit61 nearby which there existed Fcgr2b, c-jun, Fas and tumor necrosis factor (TNF)alpha, H-2 genes according to the QTL analysis. CONCLUSION: B1 cell proliferation in New Zealand mice was controlled by multiple genes and the candidate susceptibility genes were Fcgr2b, c-jun, Fas derived from NZB strain and TNF alpha, H-2 genes derived from NZW strain. PMID- 12757664 TI - [The value of reticulated platelet counts in diagnosing thrombocytopenic disorders]. AB - OBJECTIVE: To evaluate the significance of reticulated platelets (RPs) in the diagnosis of thrombocytopenic disorders and the relationship between RPs and the proliferative degree of megakaryocyte (MK) in bone marrow. METHODS: With thiazole orange as a fluorescent dye, RPs were measured by analyzing the RNA content in platelets with flow cytometry and the percent and absolute counts of RPs were calculated. RESULTS: (1) The percent and absolute counts of the RPs in a normal group were (8.4 +/- 2.5)% and (16.8 +/- 6.8) x 10(9)/L respectively. (2) As compared with the normal group, the patients with idiopathic thrombocytopenic purpura (ITP) and hypersplenism had a significantly high percent and low absolute counts of RPs (P < 0.01). In patients with aplastic anemia, both the percent and absolute counts of RPs were at low levels (P < 0.05 and P < 0.01, respectively). There was no difference of RP percentage between the patients with acute leukemia or myelodysplastic syndromes and normal controls, but the absolute counts of RPs in the former was significantly lower than that in the latter. There was no difference between the percent and absolute counts of RPs among ITP patients with different proliferative degree of MK in bone marrow. (3) In all the diseases mentioned above, it was shown that RP percentage returned to normal in the effective cases after treatment, but no such change was found in the ineffective cases. CONCLUSIONS: Reticulated platelet counts contribute to the aetiology determination of the thrombocytopenia. It is also a valuable diagnostic method and a monitoring marker. There is no relationship between reticulated platelet counts and the counts of MK proliferation in bone marrow. PMID- 12757665 TI - [Phrenic radix conduction time in inflammatory demyelinating polyneuropathy patients]. AB - OBJECTIVE: To investigate the features and application of phrenic radix conduction time (PRCT) in inflammatory demyelinating polyneuropathy (IDP) patients. METHODS: Distal motor latency (DML) was recorded by stimulating phrenic nerve electrically and phrenic nerve root magnetically in 20 IDP patients with surface electrodes at intercostal space. The difference of the two DML was calculated and compared with those recorded previously from 61 sides in 31 healthy control subjects. RESULTS: PRCT of the IDP patients was longer than that of the controls, and it correlated with the severity of clinical dyspnea. The abnormal rate of PRCT was higher than the incidence of clinical dyspnea. CONCLUSIONS: PRCT could reflect the functional status of the spinal roots related to respiration in IDP patients at an electrophysiological level. It correlates well with the degree of clinical dyspnea. Its detection is helpful to find out the subclinical dyspnea in IDP patients. PMID- 12757668 TI - Outbreak of SARS. PMID- 12757666 TI - [A retrospective clinical study of chronic high altitude disease complicated by acute exacerbation in high altitude environment]. AB - OBJECTIVE: To study chronic high altitude disease (CHAD) with concurrent acute high altitude disease (AHAD) in regions of high altitude. METHODS: 18,090 inpatients from Feb. 1956 to Dec. 1995 conforming to a screening standard were observed in a hospital located at altitude 3658 m. 1,028 inpatients suffering from CHAD when hospitalized were collected as a study group. 17,020 inpatients suffering from non-HAD when first hospitalized served as a control group. The morbidity rate of AHAD in these two groups in a follow-up period of 1 - 20 years was analyzed. RESULTS: (1) AHAD morbidity rate was increasing with prolongation of observation time in the control group, but it was not so in the study group. Annual and accumulative morbidity of AHAD in the study group was obviously higher than that in the control group (P < 0.005, OR = 5.03, RR = 4.33). (2) The morbidity rates of three types of AHAD aside from high altitude pulmonary edema (HAPE) of high altitude hypertension (HAH) group and high altitude cerebral edema (HACE) of high altitude heart disease (HAHD) group was obviously higher in the study group than in the control (P < 0.05 - 0.005). AHAD morbidity rate in HAHD group and Monge's disease was 23.5% and 22.0% (OR = 7.33 - 6.71, RR = 5.86 - 5.47). (3) AHAD morbidity rate in HAHD group and Monge's disease group was obviously higher than that in the control, constituting mainly a high morbidity of mild acute high altitude disease. CONCLUSION: The risk of AHAD increases about 5-fold in CHAD patients than in the multitude of high altitude acclimatization, being most evident in HAHD and Monge's disease. PMID- 12757669 TI - Transradial diagnostic and percutaneous coronary intervention. AB - OBJECTIVE: To describe the technique and rationale of coronary diagnostic and intervention procedures via radial artery. DESIGN: A descriptive study. PLACE AND DURATION OF STUDY: From January, 2000 to August, 2001 at National Institute of Cardiovascular Diseases (NICVD), Karachi. SUBJECTS AND METHODS: A total of 167 (3.6%) patients underwent TR approach for both diagnostic and intervention procedures. The minimum age of the patients was 20 years whereas maximum age was 75 years. All patients with positive Allen's Test were included in the study. All the procedures were done by using the right radial artery approach. RESULTS: Out of 167 cases 76% were diagnostic and 24% were in Percutaneous Coronary Intervention (PCI) group. In 3% the radial artery approach was not successful. In PCI group disease, pattern was single vessel (58%) with mostly left anterior descending artery(LAD) involvement (44%). Lesions were mostly low to moderate risk. In PCI group 51 stents both pre-mounted and un-mounted (bare) were used. In 19% cases, direct stenting was done while 12.5% patients received Abciximab and 7.5% patients underwent IVUS for lesion quantification. In PCI group, procedure was unsuccessful in 2.4% cases due to inability to cannulate and negotiate the lesion. In our limited local experience of 167 cases of TR approach, there were no major complications like major bleed, limb ischemia etc. CONCLUSION: The TR approach for invasive procedures yields comparable results to femoral approach. It has a major benefit of reduction in puncture site related complications vis-a vis intensive use of anticoagulants, antiplatelet, and fibrinolytics required for PCI. Additionally, the approach also increases patient comfort through early mobilization and reduction in cost. PMID- 12757670 TI - Frequency of anti-HCV antibodies in patients with lichen planus. AB - OBJECTIVE: To determine the frequency of anti-HCV antibodies, identify risk factors associated with HCV infection and to screen asymptomatic carriers in patients with lichen planus. DESIGN: A prospective study. PLACE AND DURATION OF STUDY: This study was carried out in the out patient department of Dermatology, Shaikh Zayed Hospital, Lahore, from October, 1999 to September, 2001. SUBJECTS AND METHODS: A total of 184 clinically diagnosed cases of lichen planus (LP) were selected for the study. Blood samples of all the patients were tested for anti hepatitis C virus antibodies (anti-HCV Ab). Polymerase chain reaction for hepatitis C virus was done in patients with positive anti-HCV Ab. Transcutaneous liver biopsy was performed in 7 patients with positive HCV-RNA. The histopathological results were evaluated using validated Metavir and Knodell scoring systems. RESULTS: Out of 184 LP patients, 43 (23.4%) were anti-HCV antibodies positive. Females were predominantly affected and male to female ratio was 1:5.1. Maximum positivity for anti-HCV was observed in age group 31-40 years (39.53%) followed by 41-50 years (25.58%). Eighty-one percent patients had history of dental treatment and 63% had received multiple injections for various ailments. Forty percent patients had family history of jaundice while 26% had jaundice in the past. Ten out of 16 anti-HCV antibody positive patients, checked for HCV-RNA, had high levels of virus in blood. Transcutaneous liver biopsy done in 7 patients revealed underlying liver disease at various stages. Four patients treated with alpha-interferon and ribazole therapy for liver disease, showed marked improvement in their skin disease. CONCLUSION: A high prevalence of HCV infection was detected in patients with lichen planus. Patients with lichen planus should be screened for HCV carrier state. PMID- 12757671 TI - Perceptions of menopause among rural women of Lahore. AB - OBJECTIVE: The present study was undertaken to document the perceptions of Pakistani women regarding menopause. DESIGN: Cross-sectional descriptive study. PLACE AND DURATION OF STUDY: The study was conducted in the suburb of Lahore from 1st July 2000 to 31st August 2000. SUBJECTS AND METHODS: The data were collected from a rural population of 28,419 individuals living in 20 villages outside Lahore. A systematic random sample of 130 women was drawn from those 1337 women, who had reached natural menopause. In-depth interviews were conducted in local dialect. RESULTS: The age of the population ranged from 42 to 80 years with a mean of 59.8+/-7.4 years. The mean age at menopause was 49.0+/-3.6 years. Majority of the women (82.3%) considered menopause as a positive change. According to 71.6% women, their relationship with the family had either improved or there was no change. There was an increase in libido among 70.5% women, decrease in libido among 5.7% and no change among 23.8% women. However, 16.9 % women reported dyspareunia. Seven percent of the women suffered a fracture after menopause. None of the study subjects reported postmenopausal bleeding. Three women (2.3%) had cardiac ailments. The proportion of women showing a positive attitude towards menopause was significantly higher (p<0.005) among our study subjects as compared to those from India, Thailand, USA and Australia. CONCLUSION: The majority of study subjects did not consider menopause as a negative milestone, loss of time, a partial death or a disease as seen in many western populations. On the contrary menopause is regarded as a natural phase in a woman's reproductive life PMID- 12757672 TI - Pregnancy with hyperthyroidism. AB - OBJECTIVE: The purpose of the study was (i) to diagnose the pregnant women suffering from hyperthyroidism by differentiating their symptoms from normal pregnancy and confirming this disease biochemically and (ii) to treat such patients medically to achieve euthyroid state of the mother while monitoring the developing fetus for better obstetrical outcome. DESIGN: A descriptive comparative study. PLACE AND DURATION OF STUDY: This was a hospital-based study carried out on the patients presented at Federal Government Services Hospital, Islamabad from 1997 to 1998. SUBJECTS AND METHODS: A total of 30 patients in reproductive age group, suffering from hyperthyroidism with pregnancy, were studied. Patients were recruited in the study in their first trimester having no associated medical problems. The patients were divided into two groups. Group I patients were diagnosed for hyperthyroidism for the first time during pregnancy by biochemical methods. These patients were treated medically. After treatment effects on their previous obstetrical complications, general health and fetal well-being were monitored. The group II patients were already under medical treatment and served as control. Their thyroid status in all trimesters, types of medicine, dose adjustment and maternal and fetal well-being were compared with the patients of group I. The patients of both the groups were treated medically with beta blockers and anti-thyroid drugs mainly carbimazole. For fetal monitoring, ultrasonography (USG) and cardiotocography (CTG) were carried out. RESULTS: The most common type of hyperthyroidism, in patients of both the groups, was toxic nodular goiter that affected 70% of the patients. The most common previous obstetrical complication in patients of both the groups was abortion (85.7%), followed by premature labour (7%). Carbimazole was the drug of choice given to 70% and 60% patients of group I and group II respectively. Having achieved euthyroid state, dose reduction was possible in 65% patients of group I and 83% patients of group II. CONCLUSION: Euthyroid state in mothers, by the time of delivery, can be easily achieved with antithyroid drugs and b blockers and the aim to improve obstetrical outcome can be successfully achieved. Hence obstetrical complications, which otherwise affect 93.33% of such patients, can easily be avoided through medical treatment. PMID- 12757673 TI - Sociocultural determinants of induced abortion. AB - OBJECTIVE: To determine the frequency of induced abortion and identify the role of sociocultural factors contributing to termination of pregnancy and associated morbidity and mortality in hospital setting. DESIGN: Prospective observational study. PLACE AND DURATION OF STUDY: The study was conducted in the Department of Obstetrics and Gynaecology, Jinnah Postgraduate Medical Centre, Karachi from January 1999 to June 2001. SUBJECTS AND METHODS: The patients who were admitted for induced abortion were interviewed in privacy. On condition of anonymity they were asked about the age, parity, family setup and relationships, with particular emphasis on sociocultural reasons and factors contributing to induction of abortion. Details of status of abortionist and methods used for termination of pregnancy, the resulting complications and their severity were recorded. RESULTS: Out of total admissions, 57(2.35%) gave history of induced abortion. All women belonged to low socioeconomic class and 59.6% of them were illiterate. Forty three (75.5%) of these women had never practiced contraception. Twenty-four (42%) were grandmultiparae and did not want more children. In 29 women (50.9%) the decision for abortion had been supported by the husband. In 25 women (43.8%) abortion was carried out by Daiyan (traditional midwives). Serious complications like uterine perforation with or without bowel injury were encountered in 25 (43.8%) of these women. During the study period illegally induced abortion accounted for 6 (10.5%) maternal deaths. CONCLUSION: Prevalence of poverty, illiteracy, grand multiparity and non-practice of contraception are strong determinants of induced abortion. PMID- 12757674 TI - Percutaneous renal biopsy by automated biopsy gun. AB - OBJECTIVE: To study the safety and efficacy of automated biopsy gun in obtaining the renal tissue by percutaneous renal biopsy. DESIGN: Single centered prospective hospital based study. PLACE AND DURATION OF STUDY: Department of Medicine, Unit II, Lahore General Hospital, Lahore for more than one year from January 2000 to February 2001. SUBJECTS AND METHODS: Forty patients of either gender having clinical features of renal disease with overt proteinuria were included in the study. An informed consent was taken from patients. Patients were lied prone, local anaesthesia was given at lower pole of left kidney. Renal biopsy was performed under ultrasound guidance, using 18G spring loaded automated biopsy gun. RESULTS: Majority of patients, 28(70%), was male. The mean age was 26.9 years with range of 5-70 years. In 37 (97.5%) patients successful biopsy was done while there was its failure in 3 (7.5%) patients. The average core length of renal tissue obtained was Mean+/-SD (1.02+/-0.34) cm with a mean number of glomeruli per core Mean+/-SD (7.76+/-2.36) and histopathological diagnosis was made in all cases. Total number of attempts were 1-3. In 70% patients single attempt was done. No major complication related to the procedure was seen. CONCLUSION: Renal biopsy with automated biopsy gun is relatively safe and effective in establishing the diagnosis of renal diseases. Although renal biopsy can be done on outpatient basis, it is better to observe the patients for 24 hours post-operatively for any complication PMID- 12757675 TI - Differentiation of pulmonary embolism from high altitude pulmonary edema. AB - OBJECTIVE: To differentiate the high altitude pulmonary edema (HAPE) from pulmonary embolism (PE) by clinical probability model of PE, lactate dehydrogenase (LDH), aspartate transaminase (AST) and D-dimer assays at high altitude. DESIGN: A prospective study. PLACE AND DURATION OF STUDY: The study was carried out at CMH, Skardu, from October 2001 to December 2002. SUBJECTS AND METHODS: Consecutive 40 patients evacuated from height > 3000 meters with symptoms of PE or HAPE were included. Clinical pretest probabilities scores of PE, Minutex D-dimer assay (Biopool international) and cardiac enzymes estimation by IFCC approved methods, were used for diagnosis. Mann-Whitney U test was applied by using SPSS and level of significance was taken at (p<0.05). RESULTS: Out of 40 subjects, 31 HAPE and 9 patients of PE were initially diagnosed on the basis of clinical features, D-dimer assay and V-Q scanning. Out of 9 patients of PE; 3 had plasma D-dimer between 250-500 ng/ml and 6 > 500 ng/ml. Plasma D-dimer of 500 ng/ml was considered as cut-off value; 6 (66.7%) patients of PE could be diagnosed and 30 (96.7%) cases of HAPE excluded indicating very good negative predictive value. Serum LDH, AST and CK were raised above the reference ranges in 8 (89%), 7 (78 %) and 3 (33 %) patients of PE as compared to 11 (35%), 6 (19%) and 9 (29 %) of HAPE respectively. CONCLUSION: Clinical assessment in combination with D-dimer assay, LDH and AST can be used for timely differentiation of PE from HAPE at high altitude where diagnostic imaging procedures are not available. PMID- 12757676 TI - Incidence of respiratory distress syndrome. AB - OBJECTIVE: To determine the incidence of respiratory distress syndrome (RDS) in hospital born babies. DESIGN: A prospective study. PLACE AND DURATION OF STUDY: The study was conducted in the neonatal intensive care unit (NICU) at the Pediatric department, in collaboration with the Obstetrics and Gynecology Department, Military Hospital, Rawalpindi, over a period of one year from January to December 2000. SUBJECTS AND METHODS: All live born infants delivered at the hospital and who fulfilled the diagnostic criteria of respiratory distress syndrome (RDS) were included in the study. RESULTS: Ninety-four neonates developed RDS. Out of these, 88 (93.61%) were preterm and 06 (6.38%) were term infants. There was a male preponderance (65.95%). RDS was documented in 1.72% of total live births, 37.28% of preterm and 0.11% of term neonates born at the hospital. The incidence of RDS was 100% at 26 or less weeks of gestation, 57.14% at 32 weeks, and 3.70% at 36 weeks. The mortality with RDS was 41 (43.61%). CONCLUSION: RDS is the commonest cause of respiratory distress in the newborn, particularly, in preterm infants. It carries a high mortality rate and the incidence is more than that documented in the Western world. PMID- 12757677 TI - Liaison psychiatry and referral rates among hospitalized patients. AB - OBJECTIVE: The study was conducted to assess the psychiatric co-morbidity among general hospital patients and their rate of referrals. DESIGN: A hospital based descriptive observational study. PLACE AND DURATION OF STUDY: The study was conducted in a private teaching hospital for a period of 6 months. METHOD: All admitted patients in different disciplines were assessed on a structured proforma supplemented by detailed psychiatric interview on positive cases. Morbidity patterns were assessed on the basis of ICD-10 classification. The total referral record was also maintained and compared with the data of morbidity and the number of referred cases. RESULTS: Out of the total 487 patients seen, 180 patients were found to have psychiatric co-morbidity; depression outnumbered all psychiatric disorders followed by mixed anxiety depression and somatoform disorders. Married female formed the majority group and the vulnerable age was between 41 to 60 years. Large segment of patients belonged to the discipline of internal medicine with the lowest (12%) referral rate. Out of 121 patients, referred from department of psychiatry, 44% were seen by the department of medicine followed by the gynecology and other disciplines. CONCLUSION: There is generally a low referral rate despite significant mental health co-morbidity. Training is needed for non-psychiatrists and there is a need for development of consultation-liaison psychiatry services in hospitals PMID- 12757678 TI - Comparison between strictureplasty and resection anastomosis in tuberculous intestinal strictures. AB - OBJECTIVE: To compare the effectiveness, safety and morbidity of strictureplasty with resection anastomosis in patients with tuberculous small gut strictures. DESIGN: Prospective comparison study. PLACE AND DURATION OF STUDY: Surgical Unit I of Rawalpindi General Hospital from March 2000 to February 2002. PATIENTS AND METHODS: Thirty patients who presented with intestinal obstruction due to tuberculous strictures, and underwent either resection anastomosis or strictureplasty were included in the study. Data was collected on a proforma and analyzed using software SPSS (version 8.0). Chi-square and t-test were used to test the hypothesis. Main outcome measures included the presence or absence of postoperative leakage anastomosis, wound infection, recurrence of intestinal obstruction and postoperative stay. RESULTS: Chi-square test applied to see the effectiveness showed no significant difference (p>0.5) between the two procedures. t-Test on the score of morbidity also showed no significant difference (p>0.5) between the two procedures. CONCLUSION: Both procedures performed were equally effective and had equal morbidity in cases of intestinal tuberculous strictures. Strictureplasty is superior to resection anastomosis in cases of multiple strictures as it conserves gut length and can even be performed safely in cases with coexistent gut perforation. PMID- 12757679 TI - Evaluation of low ligation and high ligation procedures of varicocele. AB - OBJECTIVE: To describe the mode of presentation of varicocele and to compare the low ligation (Inguinal approach/Ivanissevich's procedure) and high ligation (Retroperitoneal approach/modified Palomo's procedure) of varicocele repair regarding efficacy and postoperative complications. DESIGN: Institutional-based randomized comparative clinical trial. PLACE AND DURATION OF STUDY: This study was conducted at the Urology Department, Bolan Medical College and Sandeman Provincial Teaching Hospital, Quetta from December 1996 to November 2001 (5 years). SUBJECTS AND METHODS: A total of 213 patients of varicocele who underwent treatment in the department and completed 6 months follow-up were included in the study. The mode of presentation of varicocele was recorded for all patients. For treatment purpose patients were randomly divided into two groups: in group I (n: 115) varicocele was ligated by inguinal approach (Ivanissevich's procedure) while in group II (n: 98) by retroperitoneal approach (modified Palomo's procedure). The efficacy and postoperative complications of the two procedures were carefully recorded and compared between the two groups. RESULTS: The mean age of the patients of group-I and group-II was 27.2 and 27.5 years respectively. More than 50% patients of both groups presented with feeling of heaviness/dragging sensation on ipsilateral side. Presentation with infertility/sub fertility in group-I and group-II was 16.5% and 15.3% respectively. Majority of patients of both groups had grade III varicocele with visible deformity. The rate of postoperative complications was very low in both groups of patients. Recurrence of varicocele and postoperative hydrocele formation were slightly more common in group-II as compared to group-I but the difference was statistically not significant (p>0.05). An extremely significant improvement occurred postoperatively in both sperms density and motility in infertile/sub fertile patients of both groups. CONCLUSION: Both procedures of varicocele repair are equally effective and have a low rate of complications. There is no significant difference in postoperative complications. An extremely significant improvement occurs in semen parameters of infertile/sub fertile patients postoperatively, irrespective of method/technique of varicocelectomy PMID- 12757680 TI - Management of difficult airway by retrograde tracheal intubation. AB - A case of difficult intubation is presented in a patient of adenoid cystic carcinoma with a large right-sided facial defect. She was managed with radiotherapy and a myocutaneous flap reconstruction was done with retrograde tracheal intubation PMID- 12757681 TI - Precocious pseudopuberty due to juvenile granulosa cell tumor. AB - A case of precocious puberty occurring in a young girl is presented. Vaginal bleeding and secondary sexual characteristics had occurred at 7 years of age associated with an abdominal mass. These findings were due to a functional juvenile granulosa cell tumor in the right ovary. Right adenectomy was performed. Histopathology was confirmatory. PMID- 12757682 TI - Cerebrospinal fluid ascites. AB - Cerebrospinal fluid (CSF) ascites can be a refractory problem in the management of patients with hydrocephalus developing as a result of tuberculous meningitis. A young 17-year-old patient developed CSF ascites due to the drainage of CSF into the peritoneal cavity via a ventriculoperitoneal shunt. The ascites failed to resolve despite repeated courses of antituberculous chemotherapy. Eventually diversion of CSF away from the peritoneal cavity through a ventriculoatrial shunt relieved the ascites PMID- 12757683 TI - Colonic lipoma. AB - Lipoma of the colon is rare and may lead to intestinal obstruction. We have presented two cases of colonic lipoma. Both were elderly females, one presented with diarrhea and the other with sub-acute intestinal obstruction. After colonoscopy surgical removal was done. Histopathology revealed lipoma PMID- 12757684 TI - Vesical gossypiboma. AB - Gossypiboma (retained surgical sponge) is an iatrogenic complication. It is infrequently reported in literature because of legal implications setting up a vicious cycle of non-anticipation and misdiagnosis. Four cases of vesical gossypiboma following transvesical prostatectomy are presented. Clinical presentation, diagnostic aspects and preventive strategies are discussed. PMID- 12757685 TI - Paternalistic model of medical practice. PMID- 12757686 TI - A review of rotavirus diarrhea in Pakistan: how much do we know? AB - INTRODUCTION: Rotavirus diarrhea has a worldwide distribution, infecting almost all children by the age of 3-5 years. EPIDEMIOLOGY: A comparable etiological 2 year survey carried out by the W.H.O Diarrheal Disease control (CDD) Program in 1991, in a multicenter study in 5 developing countries including Pakistan revealed that Rotavirus was found to be the most frequently detected pathogen in diarrheal episodes, during the first year of life, with the highest incidence (20%) occurring among 6-11 months old. Two other studies done in Pakistan, in under five children done in Lahore (between 1985 and 1991) and Rawalpindi (between May 1983 and April 1984) showed that Rotavirus was the second most common Diarrhea causing enteric pathogen following E.Coli TRANSMISSION: Rotaviruses are shed in high concentrations 2 days before and as many as 10 days after onset of symptoms in immunocompetent hosts, thus being an important source of viral transmission. CLINICAL COURSE: A multicenter study in 5 developing countries including Pakistan conducted by WHO CDD program revealed that only 1.8 % of cases presented with severe dehydration and these were mostly due to Rotavirus, V. Cholerae and ETEC13. DIAGNOSTIC TESTS: A study conducted in local hospitals in Pakistan during the period of October 1985-April 1986 compared the different diagnostic modalities for the detection of rotavirus in the faeces of children with acute diarrhea. The study all methods detected Rotavirus to varying degrees but ELISA was found to be the most sensitive method with 72.4% stools being positive. PREVENTIVE STRATEGIES: A study was conducted in Lahore (Pakistan) among 72 infants 6 weeks old in 1991 to assess safety and efficacy of RRV vaccine. It was found that of all infants given RRV with OPV, 50% had a two to four-fold rise in neutralization titers against rotavirus. RRV was found to be safe and not associated with adverse reactions in the 6 weeks old infants. CONCLUSION: With regards to Pakistan, there is a great need for defining rotavirus associated disease burden and strain prevalence. We also need to conduct Rotavirus vaccine trials to assess its efficacy and safety in our setting. PMID- 12757688 TI - The efficacy of chemoprophylaxis against malaria with chloroquine plus proguanil, mefloquine, and atovaquone plus proguanil in travelers from Denmark. AB - BACKGROUND: The risk of malaria infection in travelers is seldom known in detail and neither is the efficacy of different prophylactic regimens, due to a lack of controlled trials. Surveillance of malaria diagnosed after return can provide data on risk and efficacy. METHODS: An open case-control study was initiated. Imported cases were notified to our department and were studied in 320 permanent residents in Denmark, returning from abroad with malaria from 1997 to 1999. These were compared with a group of 600 travelers who were not infected with malaria and matched by age, sex, and destination. Information on the use of chemoprophylaxis and the length of stay in malarious areas were obtained by questionnaire. RESULTS: Two hundred cases of Plasmodium falciparum malaria were notified of which 103 had used chloroquine and proguanil, 16 mefloquine, and 3 atovaquone and proguanil as prophylaxis, whereas the rest had taken other drugs or no prophylaxis. This study showed that the risk increased with increasing exposure and that compliance was lower especially for mefloquine users in malaria cases compared with controls. The study provided the first comprehensive data on the use of atovaquone/proguanil to travelers. The estimated efficacy of chloroquine and proguanil, mefloquine, and atovaquone and proguanil in fully compliant users was 1:599, 1:2,232, and 1:1,943, respectively, P. falciparum cases per prescription. The country specific risk data showed that the risk of getting malaria varied from 1 per 140 travelers to Ghana to almost 1 per 40,000 to Thailand, providing data that allow the use of prophylaxis to be restricted to high-risk areas. CONCLUSION: There was a considerable variation in risk between the countries with the highest risk in tropical Africa. Chloroquine and proguanil was less efficient compared with mefloquine. Atovaquone/proguanil (Malarone) was at least as efficient as mefloquine, but breakthroughs were observed. PMID- 12757687 TI - Meningococcal carriage in Umra pilgrims returning from Saudi Arabia. AB - BACKGROUND: Moslems from all over the world go to Mecca and Medina in Saudi Arabia for two types of pilgrimage: the major pilgrimage (hajj) and the minor (umra). An international outbreak of meningococcal disease with serogroup W-135 occurred in association with hajj pilgrimage in the years 2000 and 2001, and it has been shown that pharyngeal carriage of a single W-135 strain was high in returning hajj pilgrims. We investigated the meningococcal carriage in umra pilgrims to determine the extent of circulation of this strain, during the minor pilgrimage. METHOD: Tonsillopharyngeal swabs were taken from umra returnees. Serogrouping and pulsed field gel electrophoresis were performed on all meningococcal isolates. Subjects were questioned about the occurrence of symptoms of upper respiratory tract infection and use of antibiotics during the pilgrimage. RESULTS: were compared with those previously reported in hajj pilgrims. Results: We enrolled 160 pilgrims returning from the umra pilgrimage in 2001. The meningococcal carriage rate was 1.3%, which is significantly lower compared with the hajj pilgrimage (17%; p<0.001). None of the umra pilgrims carried serogroup W-135, whereas 90% of the isolates in returning hajj pilgrims were Neisseria meningitidis W-135. CONCLUSIONS: Meningococcal carriage during the umra pilgrimage was significantly lower compared with the hajj pilgrimage in the year 2001. No carriage of N. meningitidis W-135 was documented in umra pilgrims, whereas this was the predominant serogroup in hajj pilgrims. Public health measures to reduce the potential introduction of N. meningitidis W-135 into the countries of origin of returning pilgrims need to be prioritized for the hajj pilgrimage. PMID- 12757689 TI - Avoiding misdiagnosis of imported malaria: screening of emergency department samples with thrombocytopenia detects clinically unsuspected cases. AB - BACKGROUND: Misdiagnosis of imported malaria is not uncommon and even abnormal routine laboratory tests may not trigger malaria smears. However, blind screening of all thrombocytopenic samples might be a possible way to detect clinically unsuspected malaria cases in the accident and emergency department (AED). METHODS: The frequency and degree of thrombocytopenia was determined in two cohorts of malaria patients (Lisbon, Portugal and Berlin, Germany). The frequency of thrombocytopenia in full blood count (FBC) samples from patients presenting at the ED at a large teaching hospital in Lisbon, Portugal, was determined and compared with urgent samples from in-patients, both determined at a dedicated emergency laboratory. A cut-off value was established for screening of all FBC samples from the ED with Giemsa-stained thick-blood films. RESULTS: In 4,362 unselected samples from the ED in Lisbon a thrombocytopenia of <150,000/microL was found in 7% and of <100,000/microL in 2.2% of cases (2.5 samples/day with <100.000/microL). In patients with malaria, a thrombocytopenia was found in 75% and 77% (<150,000/microL) or 53% and 45% (<100,000/microL) in Lisbon, (n=60) and Berlin (n=170), respectively. Blind screening of all samples with <100,000 thrombocytes/microL by thick-blood film microscopy led to the diagnosis of 5 unsuspected malaria cases (3 Plasmodium falciparum, 1 Plasmodium vivax and 1 Plasmodium ovale), during the study. The diagnosis of each unsuspected malaria case would have cost 21 hours of dedicated technician's time. CONCLUSIONS: The problem of clinically unsuspected malaria seems to be more common than generally expected and is dependent on the local incidence of malaria as well as clinical and laboratory expertise. The blind screening of all thrombocytopenic samples with <100,000/microL may be a cost-effective way to reduce the misdiagnosis of imported malaria. PMID- 12757690 TI - A preliminary study on travel health issues of medical students undertaking electives. AB - BACKGROUND: With the inclusion of elective programs, often overseas, in many medical courses, it was decided that a preliminary retrospective analysis of health problems associated with these programs in medical students from the University of Tasmania would be desirable. METHODS: A questionnaire covering general travel health issues was distributed to all medical students in the University of Tasmania, on return from their elective. They were asked to complete the questionnaire and return it on an anonymous and voluntary basis. In addition, student elective submissions were consulted for information relating to their chosen destination. RESULTS: Results of the study indicate that general practitioners were the most common source of pretravel advice for Tasmanian medical students. Overall, 64% of students experienced some sort of health problem of which travelers' diarrhea was the most common. Most problems were mild and self-limiting, but a number of serious infections were recorded, including acute leptospirosis, paratyphoid, and Staphylococcus aureus cellulitis. Of particular concern were the reports of assault and sexual harassment recorded by several students. CONCLUSIONS: Elective programs are an important part of many medical courses. With the widespread destinations chosen by students, it is important that they be given adequate pretravel health advice. Most of the health problems encountered by students from the University of Tasmania were mild, but exposure to serious infections was recorded. It is thus imperative that students take out travel health insurance and that they are counseled on how to avoid dangerous situations while abroad. PMID- 12757692 TI - Managing cancer: the role of holiday taking. AB - BACKGROUND: The primary objective of this paper is to consider whether holiday taking may have a role to play post-diagnosis/treatment, within the rehabilitation process for cancer patients. METHOD: A qualitative study was undertaken. Informants consisted of cancer patients (n=24), their family members (n=13) and friends (n=2), consultants (n=2) pediatric oncology nurses (n=2), social workers (n=2), a voluntary services coordinator (n=1), and a play specialist (n=1). All were accessed through four facilitators at Christie Hospital, National Health Service Trust, Manchester, United Kingdom. Data were collected via interviews, telephone discussions, letters, personal accounts, and health diaries between January 2000 and March 2001. Questions were asked regarding the perceived effects of travel during serious illness, travel propensity, and factors inhibiting full participation. RESULTS: Benefits to health and well being, particularly of a sociopsychological nature, were identified. Such benefits impacted upon personal health, social effectiveness, personal identity, self-image, independence, future career prospects, and personal behavior. Factors inhibiting travel included the side effects of treatment, other peoples' reactions to the visible signs of cancer (e.g., hair loss, surgical scars), inflated insurance costs, a lack of holiday information sources, and communication issues (e.g., language barriers). CONCLUSIONS: Holiday taking offers a vehicle for transcending illness, even if only for a short period of time. Travel offered a range of therapeutic opportunities as well as providing a necessary means of escapism. Promoting travel as part of the rehabilitation process may well generate more intrinsic benefits than are currently appreciated. Such benefits may also be of broader application to patients facing other similarly complex illnesses. PMID- 12757691 TI - Imported schistosomiasis in Europe: sentinel surveillance data from TropNetEurop. AB - BACKGROUND: Schistosomiasis is a major parasitic disease, increasingly imported into temperate climates by immigrants from and travelers to endemic areas. METHOD: To generate valid data on imported infectious diseases to Europe and to recognize trends over time, the European Network on Imported Infectious Diseases Surveillance (TropNetEurop) was founded in 1999. Three hundred and thirty-three reports of schistosomiasis were analyzed for epidemiologic and clinical features. RESULTS: Male patients accounted for 64% of all cases. The average age of all patients was 29.5 years. The majority of patients were of European origin (53%). Europeans traveled predominantly for tourism (52%). Main reasons for travel for people from endemic areas were immigration and refuge (51%) and visits to relatives and friends (28%). The majority of infections were acquired in Africa; 92 infections were clearly attributable to Schistosoma haematobium, 130 to Schistosoma mansoni, and 4 to Schistosoma intercalatum. Praziquantel was the only treatment used. No deaths were recorded. CONCLUSION: TropNetEurop sentinel provides valuable epidemiologic and clinical data on imported schistosomiasis to Europe. PMID- 12757693 TI - Evaluation of a public health newsletter intended for travel agents. AB - OBJECTIVES: Travel agents are in a key position to encourage travelers to seek consultation in travel clinics. Since the beginning of the year 2000, a newsletter specifically designed to sensitize travel agents to travel health has been published by the public health authorities and distributed to all travel agencies in Quebec. This study was undertaken to evaluate the utilization and appreciation of the newsletter by travel agents and its impact on preventive practices. METHODS: During the autumn of 2001, a cross-sectional descriptive survey was carried out among travel agencies in Quebec. Data were collected using a self-administered questionnaire sent by fax with a postal follow-up. RESULTS: A total of 252 of the 950 travel agencies contacted (27%) answered our questionnaire. In all, 78% of respondents said their agency receives the newsletter. Among these agencies, the majority of respondents considered that the subjects discussed in the newsletter are interesting (often or in general: 96%), that the subjects and preventive recommendations for travel destinations are useful in the travel agent's practice (often or in general: 89%), and generally presented in an adequate way (96%). According to the respondents, the newsletter encouraged them, often or very often, to inform travelers about travel-related health problems (70%) or to recommend a consultation in a travel clinic (63%). The impact of the newsletter on the recommendation to consult was greater among agents having more than 10 years' experience (odds ratio [OR] 3.2). When asked about the best way to send them the newsletter, only 31% identified bulk mailing, which was the current mode of distribution. CONCLUSION: Satisfaction rate with the newsletter appears to be high among respondents who receive it. However, the low response rate to the survey may indicate that as a whole, the travel agents' interest in the newsletter is mitigated. Despite the limitations of this study, the results will allow us to modify some aspects of the publication, especially regarding its distribution. PMID- 12757694 TI - Travel health insurance: indicator of serious travel health risks. AB - BACKGROUND: Travel health risks documented by questionnaire surveys obtained (e.g., during homebound flights) are incomplete since they miss most patients who need to be repatriated. METHODS: All patient claim files were reviewed from 1997 to 1998, of the largest Swiss travel insurance company. RESULTS: Among 242 claims, 69.4% were due to illness, 30.6% due to accidents; infections were the most frequent illnesses, the extremities were the most frequently traumatized part of the body. Although the illness-to-accident ratio was 1:5 in industrialized countries and the Caribbean, it exceeded 3:0 in some developing regions. Accident proneness was noted in the first week abroad. CONCLUSION: Even if no denominator data are available, this analysis offers an insight in travel health risks, allowing comparison of the occurrences of very different, serious, health problems abroad. PMID- 12757695 TI - Trends in antimalarial drugs prescribed in Australia 1992 to 1998. PMID- 12757696 TI - Febrile Broca's aphasia: a rare presentation of typhoid fever. PMID- 12757697 TI - Mefloquine and anticoagulant interaction. PMID- 12757698 TI - Trends in antimalarial drugs prescribed in New Zealand 1993 to 1998. PMID- 12757699 TI - Gnathostomiasis without eosinophilia led to a 5-year delay in diagnosis. PMID- 12757700 TI - A major step on the road to understanding a unique posttranslational modification and its role in a genetic disease. AB - The posttranslational conversion of cysteine to C(alpha)-formylglycine in the catalytic site of mammalian sulfatases is deficient in the rare but devastating disorder multiple sulfatase deficiency (MSD). Two papers in this issue of Cell report the cloning of a gene responsible for this activity. PMID- 12757701 TI - Fusion with the fused: a new role for interleukin-4 in the building of muscle. AB - In this issue of Cell, G. Pavlath and coworkers demonstrate a novel role for Interleukin-4 (IL-4) in regulating the fusion of myoblasts with differentiated myotubes. The authors demonstrate that NFATc2 signaling in newly formed myotubes induces IL-4 expression and secretion which promotes myoblast fusion with pre existing myotubes. PMID- 12757702 TI - Graded positional information: interpretation for both fate and guidance. AB - Recent evidence indicates that gradients of the same extracellular molecules can act as both morphogens, specifying cell differentiation, and guidance cues, directing axon movement. We discuss how cells may use common mechanisms to convert graded information into discrete responses; and how extracellular signals provide coordinate systems that can be linked to highly diverse cellular outputs. PMID- 12757703 TI - Tails of intrigue: phosphorylation of RNA polymerase II mediates histone methylation. AB - Histone lysine methylation plays a key role in the organization of chromatin structure and the regulation of gene expression. Recent studies demonstrated that the yeast Set1 and Set2 histone methyltransferases are recruited to mRNA coding regions by the PAF transcription elongation complex in a manner dependent upon the phosphorylation state of the carboxy-terminal domain of RNA polymerase II. These studies define an unexpected link between transcription elongation and histone methylation. PMID- 12757705 TI - Multiple sulfatase deficiency is caused by mutations in the gene encoding the human C(alpha)-formylglycine generating enzyme. AB - C(alpha)-formylglycine (FGly) is the catalytic residue in the active site of eukaryotic sulfatases. It is posttranslationally generated from a cysteine in the endoplasmic reticulum. The genetic defect of FGly formation causes multiple sulfatase deficiency (MSD), a lysosomal storage disorder. We purified the FGly generating enzyme (FGE) and identified its gene and nine mutations in seven MSD patients. In patient fibroblasts, the activity of sulfatases is partially restored by transduction of FGE encoding cDNA, but not by cDNA carrying an MSD mutation. The gene encoding FGE is highly conserved among pro- and eukaryotes and has a paralog of unknown function in vertebrates. FGE is localized in the endoplasmic reticulum and is predicted to have a tripartite domain structure. PMID- 12757706 TI - The multiple sulfatase deficiency gene encodes an essential and limiting factor for the activity of sulfatases. AB - In multiple sulfatase deficiency (MSD), a human inherited disorder, the activities of all sulfatases are impaired due to a defect in posttranslational modification. Here we report the identification, by functional complementation using microcell-mediated chromosome transfer, of a gene that is mutated in MSD and is able to rescue the enzymatic deficiency in patients' cell lines. Functional conservation of this gene was observed among distantly related species, suggesting a critical biological role. Coexpression of SUMF1 with sulfatases results in a strikingly synergistic increase of enzymatic activity, indicating that SUMF1 is both an essential and a limiting factor for sulfatases. These data have profound implications on the feasibility of enzyme replacement therapy for eight distinct inborn errors of metabolism. PMID- 12757707 TI - Interaction of Akt-phosphorylated ataxin-1 with 14-3-3 mediates neurodegeneration in spinocerebellar ataxia type 1. AB - Spinocerebellar ataxia type 1 (SCA1) is one of several neurological disorders caused by a CAG repeat expansion. In SCA1, this expansion produces an abnormally long polyglutamine tract in the protein ataxin-1. Mutant polyglutamine proteins accumulate in neurons, inducing neurodegeneration, but the mechanism underlying this accumulation has been unclear. We have discovered that the 14-3-3 protein, a multifunctional regulatory molecule, mediates the neurotoxicity of ataxin-1 by binding to and stabilizing ataxin-1, thereby slowing its normal degradation. The association of ataxin-1 with 14-3-3 is regulated by Akt phosphorylation, and in a Drosophila model of SCA1, both 14-3-3 and Akt modulate neurodegeneration. Our finding that phosphatidylinositol 3-kinase/Akt signaling and 14-3-3 cooperate to modulate the neurotoxicity of ataxin-1 provides insight into SCA1 pathogenesis and identifies potential targets for therapeutic intervention. PMID- 12757708 TI - The pathogen-inducible nitric oxide synthase (iNOS) in plants is a variant of the P protein of the glycine decarboxylase complex. AB - A growing body of evidence indicates that nitric oxide (NO) plays important signaling roles in plants. However, the enzyme(s) responsible for its synthesis after infection was unknown. Here, we demonstrate that the pathogen-induced, NO synthesizing enzyme is a variant form of the P protein of glycine decarboxylase (GDC). Inhibitors of the P protein of GDC block its NO synthase (NOS)-like activity, and variant P produced in E. coli or insect cells displays NOS activity. The plant enzyme shares many biochemical and kinetic properties with animal NOSs. However, only a few of the critical motifs associated with NO production in animals can be recognized in the variant P sequence, suggesting that it uses very different chemistry for NO synthesis. Since nitrate reductase is likely responsible for NO production in uninfected or nonelicited plants, our results suggest that plants, like animals, use multiple enzymes for the synthesis of this critical hormone. PMID- 12757709 TI - IL-4 acts as a myoblast recruitment factor during mammalian muscle growth. AB - Skeletal muscle formation and growth require the fusion of myoblasts to form multinucleated myofibers or myotubes, but few molecules are known to regulate myoblast fusion in mammals. The transcription factor NFATc2 controls myoblast fusion at a specific stage of myogenesis after the initial formation of a myotube and is necessary for further cell growth. By examining genes regulated by NFATc2 in muscle, this study identifies the cytokine IL-4 as a molecular signal that controls myoblast fusion with myotubes. Muscle cells lacking IL-4 or the IL 4alpha receptor subunit form normally but are reduced in size and myonuclear number. IL-4 is expressed by a subset of muscle cells in fusing muscle cultures and requires the IL-4alpha receptor subunit on myoblasts to promote fusion and growth. These data demonstrate that following myotube formation, myotubes recruit myoblast fusion by secretion of IL-4, leading to muscle growth. PMID- 12757710 TI - Aging reduces proliferative capacities of liver by switching pathways of C/EBPalpha growth arrest. AB - The liver is capable of completely regenerating itself in response to injury and after partial hepatectomy. In liver of old animals, the proliferative response is dramatically reduced, the mechanism for which is unknown. The liver specific protein, C/EBPalpha, normally arrests proliferation of hepatocytes through inhibiting cyclin dependent kinases (cdks). We present evidence that aging switches the liver-specific pathway of C/EBPalpha growth arrest to repression of E2F transcription. We identified an age-specific C/EBPalpha-Rb-E2F4 complex that binds to E2F-dependent promoters and represses these genes. The C/EBPalpha-Rb E2F4 complex occupies the c-myc promoter and blocks induction of c-myc in livers of old animals after partial hepatectomy. Our results show that the age-dependent switch from cdk inhibition to repression of E2F transcription causes a loss of proliferative response in the liver because of an inability to induce E2F target genes after partial hepatectomy providing a possible mechanism for the age dependent loss of liver regenerative capacity. PMID- 12757711 TI - Apoptotic phosphorylation of histone H2B is mediated by mammalian sterile twenty kinase. AB - DNA in eukaryotic cells is associated with histone proteins; hence, hallmark properties of apoptosis, such as chromatin condensation, may be regulated by posttranslational histone modifications. Here we report that phosphorylation of histone H2B at serine 14 (S14) correlates with cells undergoing programmed cell death in vertebrates. We identify a 34 kDa apoptosis-induced H2B kinase as caspase-cleaved Mst1 (mammalian sterile twenty) kinase. Mst1 can phosphorylate H2B at S14 in vitro and in vivo, and the onset of H2B S14 phosphorylation is dependent upon cleavage of Mst1 by caspase-3. These data reveal a histone modification that is uniquely associated with apoptotic chromatin in species ranging from frogs to humans and provide insights into a previously unrecognized physiological substrate for Mst1 kinase. Our data provide evidence for a potential apoptotic "histone code." PMID- 12757713 TI - Computational modeling and experimental analysis of nonsense-mediated decay in yeast. AB - A conserved mRNA surveillance system, referred to as nonsense-mediated decay (NMD), exists in eukaryotic cells to degrade mRNAs containing nonsense codons. This process is important in checking that mRNAs have been properly synthesized and functions, at least in part, to increase the fidelity of gene expression by degrading aberrant mRNAs that, if translated, would produce truncated proteins. Using computational modeling and experimental analysis, we define the alterations in mRNA turnover triggered by NMD in yeast. We demonstrate that the nonsense containing transcripts are efficiently recognized, targeted for deadenylation independent decapping, and show NMD triggered accelerated deadenylation regardless of the position of the nonsense codon. We also show that 5' nonsense codons trigger faster rates of decapping than 3' nonsense codons, thereby providing a mechanistic basis for the polar effect of NMD. Finally, we construct a computational model that accurately describes the process of NMD and serves as an explanatory and predictive tool. PMID- 12757712 TI - The zipper model of translational control: a small upstream ORF is the switch that controls structural remodeling of an mRNA leader. AB - Transport of the essential amino acids arginine and lysine is critical for the survival of mammalian cells. The adaptive response to nutritional stress involves increased translation of the arginine/lysine transporter (cat-1) mRNA via an internal ribosome entry site (IRES) within the mRNA leader. Induction of cat-1 IRES activity requires both translation of a small upstream open reading frame (uORF) within the IRES and phosphorylation of the translation initiation factor eIF2alpha. We show here that translation of the upstream ORF unfolds an inhibitory structure in the mRNA leader, eliciting a conformational change that yields an active IRES. The IRES, whose activity is induced by amino acid starvation, is created by RNA-RNA interactions between the 5' end of the leader and downstream sequences. This study suggests that the structure of the IRES is dynamic and regulation of this RNA structure is a mechanism of translational control. PMID- 12757716 TI - Synthesis and in vitro trypanocide activity of several polycyclic drimane-quinone derivatives. AB - The Diels-Alder reaction between two polygodial-derived dienes and simple quinones to yield substituted naphtho- and anthraquinones, is described. The in vitro trypanocide activity for the series was determined. Two of the new compounds showed an activity ten and two times higher, respectively, than nifurtimox and benznidazole, the medicines of choice for the treatment of the acute Chagas' disease. PMID- 12757717 TI - New unusual iridoids from the leaves of noni (Morinda citrifolia L.) show inhibitory effect on ultraviolet B-induced transcriptional activator protein-1 (AP-1) activity. AB - A novel iridoid dimer in whose structure the two iridoid units are connected by a rare ether group, together with two new unusual iridoids showing significant inhibition of UVB-induced Activator Protein-1 (AP-1) activity in cell cultures, have been isolated from the leaves of noni (Morinda citrifolia L.). Their structures were determined on the basis of detailed high-field 1D and 2D spectral analysis. Their inhibitory effect on UVB-induced transcriptional Activator Protein-1 (AP-1) activity are also discussed. PMID- 12757718 TI - Specific alkylation of human telomere repeats by hairpin pyrrole-imidazole polyamide. AB - A novel hairpin polyamide-cyclopropapyrroloindole (CPI) conjugate PyImImIm-gamma PyPyPyLDu86 (conjugate 11), which targets human telomere repeats d(TTAGGG)(n)/d(CCCTAA)(n), was synthesized. High resolution denaturing polyacrylamide gel electrophoresis using 44 bp DNA fragments and HPLC product analysis of a synthetic nonanucleotide demonstrated that conjugate 11 alkylates the target adenine in the telomere repeats, 5'-CCCTAA-3'. Examination of the antitumor activity of conjugate 11 using a panel of 39 cancer cell lines demonstrated that the average concentration of conjugate 11 required for 50% growth inhibition was 5.75 microM, which is superior to pepleomycin and bleomycin and comparable to cisplatin. PMID- 12757719 TI - Cancer preventive potential of trichothecenes from Trichothecium roseum. AB - Bioassay-guided separation of extracts from the culture broth and mycelium of the fungus Trichothecium roseum, aiming at the discovery for cancer preventive agents, resulted in the isolation of three new trichothecene sesquiterpenes, trichothecinols A-C (1-3) together with three known analogues, trichothecin (4), trichodermol (5) and trichothecolone (6). Compounds 1-6 exhibited remarkably potent inhibition against Epstein-Barr virus early antigen (EBV-EA) activation induced by the tumor promoter, 12-O-tetradecanoylphorbol-13-acetate (TPA). Further compound 1 strongly inhibited TPA-induced tumor promotion on mouse skin initiated with 7,12-dimethylbenz[a]anthracene (DMBA) in two-stage carcinogenesis tests. These results suggest that compound 1 might be a valuable lead for further evaluation as a cancer preventive agent. In addition to their cancer preventive activity, compound 2 was found to show modest antifungal activity against Crypotcoccus albidus and Saccharomyces cerevisiae. PMID- 12757720 TI - N-[18F]fluoroethyl-4-piperidyl acetate ([18F]FEtP4A): A PET tracer for imaging brain acetylcholinesterase in vivo. AB - N-[(18)F]Fluoroethyl-4-piperidyl acetate ([(18)F]FEtP4A) was synthesized and evaluated as a PET tracer for imaging brain acetylcholinesterase (AchE) in vivo. [(18)F]FEtP4A was previously prepared by reacting 4-piperidyl acetate (P4A) with 2-[(18)F]fluoroethyl bromide ([(18)F]FEtBr) at 130 degrees C for 30 min in 37% radiochemical yield using an automated synthetic system. In this work, [(18)F]FEtP4A was synthesized by reacting P4A with 2-[(18)F]fluoroethyl iodide ([(18)F]FEtI) or 2-[(18)F]fluoroethyl triflate ([(18)F]FEtOTf in improved radiochemical yields, compared with [(18)F]FEtBr under the corresponding condition. Ex vivo autoradiogram of rat brain and PET summation image of monkey brain after iv injection of [(18)F]FEtP4A displayed a high radioactivity in the striatum, a region with the highest AchE activity in the brain. Moreover, the distribution pattern of (18)F radioactivity was consistent with that of AchE in the brain: striatum>frontal cortex>cerebellum. In the rat and monkey plasma, two radioactive metabolites were detected. However, their presence might not preclude the imaging studies for AchE in the brain, because they were too hydrophilic to pass the blood-brain barrier and to enter the brain. In the rat brain, only [(18)F]fluoroethyl-4-piperidinol ([(18)F]FEtP4OH) was detected at 30 min postinjection. The hydrolytic [(18)F]FEtP4OH displayed a slow washout and a long retention in the monkey brain until the PET experiment (120 min). Although [(18)F]FEtP4A is a potential PET tracer for imaging AchE in vivo, its lower hydrolytic rate and lower specificity for AchE than those of [(11)C]MP4A may limit its usefulness for the quantitative measurement for AchE in the primate brain. PMID- 12757721 TI - Conformationally constrained analogues of diacylglycerol (DAG). Effect on protein kinase C (PK-C) binding by the isosteric replacement of sn-1 and sn-2 esters in DAG-lactones. AB - In order to determine the importance of the two ester pharmacophores in high affinity, conformationally constrained DAG-lactones (Lac-1-5) as PK-C ligands, we have independently replaced the sn-1 and sn-2 carbonyl esters in these compounds by ketone (2, 10, 11), amide (3, 25-28), and hydroxyl (12, 13) isosteres. Although the ketone analogue of the sn-1 ester (2) exhibited comparable activity to the parent Lac-1 when taking into account the difference in lipophilicities, the other isosteres were significantly poorer PK-C alpha ligands compared to the parent DAG-lactones. This study demonstrates that the ester functionality in DAG lactone plays an important role in the ligand's capacity to form a strong hydrogen bond with Gly253 at the active site. The discrete K(i) analysis from the sn-1 and sn-2 isosteres further confirms that the DAG-lactones bind preferentially to the C1-domain in the sn-2 binding mode, as previously suggested. PMID- 12757722 TI - 1H-Imidazo[4,5-c]quinoline derivatives as novel potent TNF-alpha suppressors: synthesis and structure-activity relationship of 1-, 2-and 4-substituted 1H imidazo[4,5-c]quinolines or 1H-imidazo[4,5-c]pyridines. AB - Structural modification of imiquimod (1), which is known as an interferon-alpha (IFN-alpha) inducer, for the aim of finding a novel and small-molecule tumor necrosis factor-alpha (TNF-alpha) suppressor and structure-activity relationship (SAR) are described. Structural modification of a imiquimod analogue, 4-amino-1 [2-(1-benzyl-4-piperidyl)ethyl-1H-imidazo[4,5-c]quinoline (2), which had moderate TNF-alpha suppressing activity without IFN-alpha inducing activity, led to a finding of 4-chloro-2-phenyl-1-[2-(4-piperidyl)ethyl]-1H-imidazo[4,5-c]quinoline (10) with potent TNF-alpha suppressing activity. The relation between conformational direction of 2-(4-piperidyl)ethyl group at position 1 and TNF alpha suppressing activity is also demonstrated by NMR. PMID- 12757723 TI - Acyl sulfonamides as potent protease inhibitors of the hepatitis C virus full Length NS3 (protease-helicase/NTPase): a comparative study of different C terminals. AB - Synthesis and inhibitory potencies of three types of protease inhibitors of the hepatitis C virus (HCV) full-length NS3 (protease-helicase/NTPase) are reported: (i) inhibitors comprising electrophilic serine traps (pentafluoroethyl ketones, alpha-keto acids, and alpha-ketotetrazoles), (ii) product-based inhibitors comprising a C-terminal carboxylate group, and (iii) previously unexplored inhibitors comprising C-terminal carboxylic acid bioisosteres (tetrazoles and acyl sulfonamides). Bioisosteric replacement with the tetrazole group provided inhibitors equally potent to the corresponding carboxylates, and substitution with the phenyl acyl sulfonamide group yielded more potent inhibitors. The hexapeptide inhibitors Suc-Asp-D-Glu-Leu-Ile-Cha-Nva-NHSO(2)Ph and Suc-Asp-D-Glu Leu-Ile-Cha-ACPC-NHSO(2)Ph with K(i) values of 13.6 and 3.8 nM, respectively, were approximately 20 times more potent than the corresponding inhibitors with a C-terminal carboxylate and were comparable to the carboxylate-based inhibitor containing the native cysteine, Suc-Asp-D-Glu-Leu-Ile-Cha-Cys-OH (K(i)=28 nM). The acyl sulfonamide group constitutes a very promising C-terminal functionality that allows for prime site optimization. PMID- 12757724 TI - Three-dimensional quantitative structure-activity relationship (3D-QSAR) studies of tricyclic oxazolidinones as antibacterial agents. AB - Oxazolidinones exemplified by eprezolid and linezolid are a new class of antibacterials that are active against Gram positive and anaerobic bacteria including methicillin-resistant Staphylococcus aureus (MRSA), methicillin resistant Staphylococcus epidermidis (MRSE) and vancomycin resistant enterococci (VRE). In an effort to have a better antibacterial agent in the oxazolidinone class, we have performed three-dimensional quantitative structure-activity relationship (3D-QSAR) studies for a series of tricyclic oxazolidinones. 3D-QSAR studies were performed using the Comparative Molecular Field Analysis (CoMFA) and Comparative Molecular Similarity Indices Analysis (CoMSIA) procedures. These studies were performed using 42 compounds; the QSAR model was developed using a training set of 33 compounds. The predictive ability of the QSAR model was assessed using a test set of 9 compounds. The predictive 3D-QSAR models have conventional r(2) values of 0.975 and 0.940 for CoMFA and CoMSIA respectively; similarly, cross-validated coefficient q(2) values of 0.523 and 0.557 for CoMFA and CoMSIA, respectively, were obtained. The CoMFA 3D-QSAR model performed better than the CoMSIA model. PMID- 12757725 TI - Synthesis and antiproliferative activity of basic thioanalogues of merbarone. AB - Three series of 5-substituted 1,3-diphenyl-6-(omega-dialkyl- and omega-cyclo aminoalkyl)thio-2-thiobarbiturates (11-13) were synthesized as polysubstituted thioanalogues of merbarone, a topoisomerase II inhibitor acting on the catalytic site. To better understand pharmacophore requirements, a forth series of conformationally constrained analogues 14 was also prepared. Derivatives 11b,e, 14b,e,h,i,j were active in the low micromolar concentration range (IC(50): 3.3 4.3 microM), whereas compounds 11a,c,d,f,h,j and 13a,b,d,g,j and 14a,d,f showed IC(50) values between 10 and 15.5 microM. In contrast, compounds 12a-c,g-j, 13e,f,h and 14k were inactive. Cytotoxicity data provided from N.C.I. on selected compounds provided evidence that 11b,d, 13d,g and 14b,d,f,h,i,j were endowed with potent antiproliferative activity against leukemia and prostate cell lines (GI(50) up to 0.01 microM). In general, bicyclic derivatives 14 were up to 10 fold more potent than monocyclic counterparts against solid tumor-derived cell lines. SAR studies indicated that, in general, a certain tolerability in length of the alkyl side chains and in shape of distal amines is allowed in the four series, but in the monocyclic derivatives (11-13) antiproliferative activity was strongly affected by the nature of the 5-substituents (COOC(2)H(5)>COCH(3)>>C(6)H(5)). Compounds 11b and 14b were also evaluated against KB cell subclones expressing altered levels of topoisomerases or the multidrug resistance phenotype (MDR). In both cases the above compounds showed a decrease in potency. In enzyme assays, 11b and 14b turned out to be inhibitors of topoisonerase II as merbaron. PMID- 12757726 TI - Solid-phase synthesis of a pepticinnamin E library. AB - Pepticinnamin E is a naturally occurring bisubstrate inhibitor of farnesyltransferase. Based on the structure of the natural product, a compound library was synthesized by variation of eight structural parameters. Following three different routes, a total of 51 analogues was synthesized on the polymeric support in 6-11-step parallel syntheses. Overall yields ranged from 3 to 63%, and the compounds were obtained with >90% purity. PMID- 12757727 TI - Identification of mono- and bisubstrate inhibitors of protein farnesyltransferase and inducers of apoptosis from a pepticinnamin E library. AB - A library of 51 analogues of the naturally occurring protein farnesyltransferase inhibitor pepticinnamin E was investigated biologically. Several compounds with pronounced inhibitory activity were discovered with the lowest IC(50) value reaching 1 microM. The library contains inhibitors which are competitive to either farnesylpyrophosphate or the peptide substrate and a bisubstrate inhibitor. This activity is supported and rationalized by molecular modelling experiments and different binding modes of the inhibitors deduced from them. Several compounds induced apoptosis in a Ras-transformed tumour cell line, and in one case this correlated with farnesyltransferase-inhibiting activity. PMID- 12757728 TI - Bisquaternary dimers of strychnine and brucine. A new class of potent enhancers of antagonist binding to muscarinic M2 receptors. AB - Bisquaternary dimers of strychnine and brucine were synthesized and their allosteric effect on muscarinic acetylcholine M(2) receptors was examined. The compounds retarded the dissociation of the antagonist [(3)H]N-methylscopolamine ([(3)H]NMS) from porcine cardiac cholinoceptors. This action indicated ternary complex formation. All compounds exhibited higher affinity to the allosteric site of [(3)H]NMS-occupied M(2) receptors than the monomeric strychnine and brucine, while the positive cooperativity with NMS was fully maintained. SAR studies revealed the unchanged strychnine ring as an important structural feature for high allosteric potency. PMID- 12757729 TI - Tripartate poly(ethylene glycol) prodrugs of the open lactone form of camptothecin. AB - Two PEG prodrugs utilizing conjugation of PEG through the C-21 acid functionality as well as the C-17 OH group of CPT hydroxy-amide open forms were synthesized and characterized. Both of these open lactone tripartate prodrugs were shown to be water soluble and highly effective in MX-1 mouse xenograph studies. Indirect evidence implies that the initial ester or carbonate bond breaking is esterase mediated in the first step of the cascade of CPT release. PMID- 12757730 TI - Synthesis of 1-D- and 1-L-myo-inosityl 2-N-acetamido-2-deoxy-alpha-D glucopyranoside establishes substrate specificity of the Mycobacterium tuberculosis enzyme AcGI deacetylase. AB - Mycothiol (MSH, 1-D-myo-inosityl 2-(N-acetyl-L-cysteinyl)amido-2-deoxy-alpha-D glucopyranoside) is the principal low molecular weight thiol in actinomycetes. The enzyme 1-D-myo-inosityl 2-N-acetamido-2-deoxy-alpha-D-glucopyranoside deacetylase (AcGI deacetylase) is involved in the biosynthesis of MSH and forms the free amine 1-D-myo-inosityl 2-amino-2-deoxy-alpha-D-glucopyranoside, which is used in the third of four steps of MSH biosynthesis. Here, we report the synthesis of two isomers of AcGI, which contain either 1-L-myo-inositol or 1-D myo-inositol. These synthetic products were used to investigate substrate specificity of the Mycobacterium tuberculosis enzyme AcGI deacetylase. PMID- 12757731 TI - Helical templating of oligopeptides by cyclodextrin dimers. AB - beta-cyclodextrin-based receptors were synthesized and tested for their ability to induce a helical fold in peptides bearing hydrophobic amino acid residues in the i, i+11- or i, i+14-positions. Circular dichroism experiments revealed that a dimeric beta-cylodextrin receptor synthesized from a [1,1'-biphenyl]-4,4'-dithiol core demonstrated an ability to fold a designed peptide bearing the artificial amino acid L-p-t-butylphenylalanine in the i, i+11-positions, while other dimeric and monomeric receptors failed to do so. Titration studies were performed using both circular dichroism and calorimetry, the analysis of which yielded an apparent K(a) on the order of 10(4)-10(5) M(-1). However, no evidence could be obtained for helical folding with a peptide carrying tryptophan residues in place of the p-t-butylphenylalanine units. Our studies suggest that receptors of this type may be useful in molecular recognition of hydrophobic, already alpha-helical peptides in aqueous solution. PMID- 12757732 TI - Soluble proteolytic enzyme release by naive and HIV-infected cultured T-cells. AB - The possible hydrolysis of leucine enkephalin was measured in the presence of cell-free supernatants obtained from naive and chronically HIV-infected immunocompetent cell lines. The data obtained indicate that, under all conditions examined, leu-enkephalin was partially degraded; its disappearance was associated with the appearance of peptides whose composition is consistent with the involvement of three enzyme classes, i.e. aminopeptidases, dipeptidylaminopeptidases and dipeptidylcarboxypeptidases. In the presence of supernatants obtained from infected cells, substrate hydrolysis was less than that measured in naive controls. This appears to result from infection-associated variations in the activity of all three enzyme classes active on the substrate, variations that were different for each class. Specifically, in unfractionated supernatants, the activity of aminopeptidases was reduced, that of dipeptidylaminopeptidase was increased, and the activity of dipeptidylcarboxypeptidases was nearly unmodified. Data obtained upon chromatographic separation of the soluble supernatants allowed for the identification of features that can be interpreted as indicating the existence of infection-associated variations in the activity of single enzymes. The sum of the data shown makes it possible to advance the hypothesis that the infection associated modifications in the release of proteolytic enzymes may contribute to the alterations in the functionality of immunocompetent cells induced by viral infection. PMID- 12757733 TI - Potential involvement of monocyte chemoattractant protein (MCP)-1/CCL2 in IL-4 mediated tumor immunity through inducing dendritic cell migration into the draining lymph nodes. AB - We previously observed that IL-4 gene transduction into a mouse colon 26 adenocarcinoma cell line abrogated its tumorigenicity due to the generation of anti-tumor CTL. DEC-205- and CD11c-double positive cells were increased in the lymph nodes of mice injected with IL-4-transfected cells between 2 and 3 days after the tumor injection, compared with those injected with parental cells. Most of these double positive cells expressed CD86 antigen. Among the chemokines with chemotactic activities against dendritic cells, monocyte chemoattractant protein (MCP)-1/CCL2, ABCD-1/CCL22, and liver and activation-regulated chemokine (LARC)/CCL20 gene expression was enhanced no later than 3 days after the tumor injection, in the draining lymph nodes of IL-4-transfected cell bearing mice. Moreover, gene expression of the receptor for MCP-1/CCL2, CCR2, was enhanced in the draining lymph nodes of the mice injected with IL-4-transfected cells, and most DEC-205-positive cells in the lymph nodes expressed CCR2. Finally, the administration of anti-MCP-1/CCL2 antibodies retarded the rate of tumor regression in mice injected with IL-4-tranfected cells, concomitantly with a decrease in DEC-205- and CD11c-double positive cell number in the draining lymph nodes. Thus, locally produced MCP-1/CCL2 may be responsible for IL-4-mediated tumor rejection presumably based on the induction of dendritic cell migration into the draining lymph nodes. PMID- 12757735 TI - Immunomodulating principles from shark cartilage. Part 1. Isolation and biological assessment in vitro. AB - Extracts from shark cartilage exhibiting powerful immunostimulating activity in vitro are described. The study shows that a simple extraction with water is very effective in producing the immunostimulating principles and implies that it is ideal for scale-up and manufacturing on a large scale. The extracts are potent stimulators of B cells and macrophages isolated from BALB/c mice spleen while stimulation of T cells was insignificant in our in vitro models. The study demonstrates that the active principles are thermally stable proteoglycans with molecular masses exceeding 100 kDa. This in vitro study represents an important step needed for further assessment of the products in vivo and their value for nutraceutical use. PMID- 12757734 TI - Anti-tumor immune response induced by the fractions derived from OK-432, a streptococcal preparation, by using a monoclonal antibody TS-2 that neutralizes the interferon-gamma-inducing activity of OK-432: comparison between the TS-2 binding and TS-2-unbinding fraction. AB - We have previously isolated a lipoteichoic acid (LTA)-related molecule (OK-PSA) from OK-432, a streptococcal agent, by affinity chromatography on a CNBr activated Sepharose 4B bound TS-2 monoclonal antibody (mAb) that neutralizes the interferon (IFN)-gamma-inducing activity of OK-432. In the current study, we compared the cytokine-inducing and anti-tumor activities of OK-PSA, a TS-2 binding fraction, with those of OK-PTF, a TS-2-unbinding fraction, in order to determine the efficacy of OK-PSA for clinical use in affinity chromatography using TS-2. In the in vitro experiments using human peripheral blood mononuclear cells (PBMCs), OK-PSA markedly induced Th1-type cytokines, while interleukin (IL) 6 and IL-10, Th2-type cytokines, were induced by OK-PTF. Th1-cytokine induction by OK-PTF was not dose-dependent and was suppressed when PBMCs were treated with a high concentration of OK-PTF. In a mouse model, Th1 cytokines were also induced by OK-PSA and Th2 cytokines were induced by OK-PTF. Th2 cytokine-inducing activity of OK-PTF was accelerated in tumor-bearing mice relative to that in healthy mice. Although the anti-tumor effect of OK-PTF was statistically significant, it was much weaker than that of OK-PSA. A significant difference between the anti-tumor effect of OK-PSA and that of OK-PTF was observed (P<0.05). Finally, OK-PSA elicited its cytokine-inducing effect via Toll-like receptor (TLR) 4, whereas OK-PTF-induced signaling was mediated by both TLR2 and TLR4. These findings strongly suggested that the affinity chromatography using TS-2 is a useful strategy to separate the effective component for cancer therapy (OK-PSA) from other components. PMID- 12757736 TI - Sodium arsenite retards proliferation of PHA-activated T cells by delaying the production and secretion of IL-2. AB - Arsenic is a metalloid that commonly contaminates drinking water, and is a known human carcinogen. It has been shown that peripheral blood mononuclear cells (PBMCs) from healthy donors treated in vitro with NaAsO(2) and stimulated with phytohemagglutinin (PHA) show a lower proliferation than nontreated cells. We reported previously a reduction in the secretion of IL-2 in NaAsO(2)-treated PBMCs stimulated with PHA, an observation that might explain, in part, the reduction in proliferation. Since arsenic induces cytoskeleton alterations, which in turn may affect protein transport of the cell, we assumed that NaAsO(2) induced an accumulation of IL-2 inside the cells, and thus a reduction in the secretion of IL-2. In order to demonstrate this hypothesis, we assessed the intracellular IL-2 at the single cell level by flow cytometry, and unexpectedly found a reduction in the percentage of IL-2 producing T cells in the presence of NaAsO(2). We tracked the proliferation of T cells by using the 5,6 carboxyfluorescein diacetate succinimidyl ester (CFSE) dye and found that NaAsO(2) slows down the entrance to cell division and delays the proliferation of cells that have already entered the cell cycle. Nevertheless, the expression of the activation molecules, CD25 and CD69, was unaltered. Assessment of the intracellular and secreted IL-2 in kinetic experiments showed that in fact, NaAsO(2) delays the production of IL-2, given that a recovery of both intracellular and secreted IL-2 was detected at 72 h. Evaluation of the cell cycle showed a higher proportion of cells in G(0)/G(1) and a lower proportion in G(2)/M in the presence of NaAsO(2). We thus conclude that NaAsO(2) reduces proliferation of T cells by delaying the production and secretion of IL-2, thus blocking T cells in G(1); as a consequence, the entry to cell cycle and the rounds of cell division are retarded, and a lower proliferation of T cells is hence observed. PMID- 12757737 TI - Characterization of B cell membrane receptors of polysaccharide isolated from the root of Acanthopanax koreanum. AB - We investigated the immunomodulatory activity of polysaccharide isolated from the root of Acanthopanax koreanum (AK) at the cellular level. AK directly increased B cell proliferation and antibody production, but did not affect the expression of IL-2, IFN-gamma or IL-4 by T cells, or T cell proliferation in vitro. Since AK cannot penetrate cells due to its large molecular mass, B cell activation may be caused by the surface binding of AK to B cell-specific receptors. The role of TLR4 as an AK receptor was shown by the fact that AK activity in B cells from C3H/HeJ mice, which are known to have a defective Toll-like receptor (TLR)-4, was found to be reduced compared with that in control cells from C3H/HeN mice. AK activity was also reduced by antibodies blocking TLR2, TLR4, CD19 or CD79b, but not by an antibody blocking CD38, which suggests AK receptor profiling in B cells. Two main differences between AK and lipopolysaccharide (LPS) were observed. First, LPS activity was inhibited by antibodies to either TLR2 or TLR4, but not by antibodies to CD19, CD79b or CD38. Another was that LPS-induced B cell proliferation was inhibited by polymyxin B (PMB), a specific inhibitor of LPS, whereas AK activity was not affected. Taken together, our results demonstrate that AK directly activates B cells, but not T cells, and suggest that AK has a broader receptor profile than LPS in B cells. PMID- 12757739 TI - Methionine enkephalin suppresses metabolic activity of a leukemic cell line (NALM 1) and enhances CD10 expression. AB - NALM-1 cells (a cell line derived from human pre-B leukemia) were exposed to the opioid pentapeptide methionine-enkephalin (Met-enkephalin) and/or to thiorphan, an inhibitor of the enzyme that degrades the enkephalins (membrane endopeptidase EC 3.4.24.11, CALLA, the CD10 marker). Metabolic and proliferative activity was assessed after 6, 24 and 48 h in microplates using a colorimetric assay with vital dye MTT. CD10 expression was determined by means of semi-quantitative RT PCR. Exposure to the Met-enkephalin at concentrations of 10(-8)-10(-6) M for 6 h reduced the MTT-activity, and after 24 and 48 h the suppression waned. Thiorphan (5 x 10(-6) M) abrogated the suppressive effect of the enkephalin, and after 6 h converted suppression into stimulation. Met-enkephalin (10(-6) M) increased and thiorphan (2.5 x 10(-6)-10(-6) M) decreased expression of CD10 at the RNA level. Suppression of the MTT uptake was attributed to the products of Met-enkephalin degradation caused by the enzymatic activity of CD10. PMID- 12757738 TI - The use of selective pharmacological inhibitors to delineate signal transduction pathways activated during complement receptor-mediated degranulation in chicken heterophils. AB - Complement receptors (CRs), along with Fc receptors, play a primary role in the removal of bacterial pathogens in poultry. The binding of serum-opsonized bacteria to CR results in the secretion of both toxic oxygen metabolites and antibacterial granules. We have previously shown that the stimulation of chicken heterophils with serum-opsonized Salmonella enteritidis induced tyrosine kinase dependent phosphorylation regulated degranulation. In the present studies, we used selective pharmacological inhibitors to investigate the roles of protein tyrosine kinases, phospholipases C and D (PLC and PLD), phosphatidylinositol 3' kinase (PI3-K), and the super family of mitogen-activated protein kinases (MAPKs) on CR-mediated heterophil degranulation. Inhibitors of receptor-linked tyrosine kinases (the tryphostins AG1478 and AG1296) had no attenuating effects on CR mediated degranulation. However, PP2, a selective inhibitor of the src family of protein tyrosine kinases, and piceatannol, an inhibitor of Syk tyrosine kinases, both significantly attenuated the CR-mediated degranulation. Additionally, the specific inhibitors of PLC, U73122, and PI3-K, LY294002, significantly decreased CR-mediated heterophil degranulation. Two inhibitors of PLD-mediated signaling, 2,3-diphosphoglycerate (2,3-DPG) and 1-butanol, hindered degranulation. Addition of purified PLD restored control levels of degranulation in heterophils in which PLD was inhibited. Lastly, SP600125, a selective inhibitor of c-Jun N-terminal kinase (JNK), inhibited degranulation; whereas neither PD98059, the inhibitor of p38 MAPK, nor SB203580, the inhibitor of extracellular signal-regulated kinase, had any effect on CR-mediated heterophil degranulation. These studies demonstrate that CRs on chicken heterophils lack intrinsic tyrosine kinase activity, but that binding of serum-opsonized bacteria activates both proximal tyrosine kinases (src and Syk kinases), but differentially activates downstream tyrosine kinases (JNK, but not p38 nor ERK). Activation of src and Syk kinases plays a significant role in signal transduction of heterophil degranulation probably by stimulating downstream phosphorylation of PLC, PLD, and PI3-K. PI3-K has also been recently shown to be an upstream mediator of JNK activation, suggesting that this enzyme can induce signaling as both a lipid kinase and protein kinase. Engaging CRs on chicken heterophils activates a proximal tyrosine kinase (src and Syk kinases)- >PLC (PLD)-->PI3-K-->JNK signal transduction pathway that induces degranulation. PMID- 12757740 TI - Combined treatment with colchicine and Herba Taraxaci (Tarazacum mongolicum Hand. Mazz.) attenuates Behcet's disease-like symptoms in mice and influences the expressions of cytokines. AB - Herbal medicine, Herba Taraxaci (Tarazacum mongolicum Hand.-Mazz.), was administered to mice with Behcet's disease (BD)-like symptoms induced by herpes simplex virus (HSV). BD is a chronic recurrent inflammatory disease. Herba Taraxaci (6 mg) was administered alone or in combination with 2 microg of colchicine to BD-like mice. Colchicine is a drug that is widely used as a medication for BD patients. The water extracts of Herba Taraxaci were administered orally once per day for 20 days. Eighty percent (8 of 10) of mice treated with Herba Taraxaci combined with colchicine showed improvement in mucocutaneous symptoms compared to 0% (0 of 10) of the nontreated group and 30% (3 of 10) treated with colchicine alone. Cytokine expression in spleen tissue collected from treated mice was analyzed by RT-PCR and FACS. Treatment with Herba Taraxaci induced IL-4 mRNA, and spleen from mice receiving the combined treatment (Herba Taraxaci and colchicines) showed an increased number of splenocytes staining with anti-IL-10 (46.8+/-6.80) compared to Herba Taraxaci (35.4+/-2.17) (p<0.05) or colchicine alone (26.2+/-4.47) (p<0.001). These results suggest that the Herba Taraxaci may be an effective complementary agent in the treatment of BD. PMID- 12757742 TI - Modulation of IL-12 p35 and p40 promoter activity by smokeless tobacco extract is associated with an effect upon activation of NF-kappaB but not IRF transcription factors. AB - Interleukin (IL)-12 (p70), composed of p35 and p40 subunits, stimulates cellular immunity and inflammation. Stimulation of IL-12 production by smokeless tobacco extract (STE) could increase the chances of oral inflammatory disease. However, p40 forms homodimers and is part of IL-23 heterodimers. Expression of p35 and p40 in response to lipopolysaccharide (LPS) and interferon (IFN)-gamma requires activation of nuclear factor-kappa-B (NF-kappaB) and interferon regulatory factor (IRF) transcription factors. To determine the impact of STE on expression of p35 and p40, the activities of p35 and p40 promoter reporter plasmids in RAW264.7 cells stimulated with STE alone or in the presence of IFN-gamma and LPS were assessed. In addition, nuclear localizations of NF-kappaB p50, p65 and IRF-1, -2 and -8 in RAW264.7 cells treated with STE were evaluated. The results show that STE alone stimulates p40 and p35 promoter activity and enhances IFN-gamma-induced p40 and p35 promoter activity. In contrast, STE had no effect on LPS-induced p35 and p40 promoter activity and diminished IFN-gamma/LPS-induced p35 promoter activity. STE had little effect upon nuclear localization of IRFs, but it stimulated nuclear localization of both NF-kappaB p50 and p65. STE also stimulated IFN-gamma-induced activation of NF-kappaB p50 but reduced nuclear localization of IFN-gamma- and IFN-gamma/LPS-induced NF-kappaB p65. SN50, an inhibitor of NF-kappaB nuclear localization, significantly lowered STE-induced p35 and p40 promoter activity. These results suggest that STE stimulation of bioactive IL-12 production is correlated with its impact upon both p35 and p40 and can be attributed in part through an effect upon NF-kappaB p50 nuclear localization. PMID- 12757741 TI - A water-extract of the Korean traditional formulation Geiji-Bokryung-Hwan reduces atherosclerosis and hypercholesteremia in cholesterol-fed rabbits. AB - Geiji-Bokryung-Hwan (GBH), a drug preparation consisting of five herbs of Cinnamomi Ramulus (Geiji), Poria Cocos (Bokryun), Mountan Cortex Radicis (Mokdanpi), Paeoniae Radix (Jakyak) and Persicae Semen (Doin), is a traditional Korean herbal medicine that is widely used in the treatment of atherosclerosis related disorders. A water extract of GBH was found to scavenge 1,1-diphenyl-2 picrylhydrazyl (DPPH) radicals and inhibit low-density lipoprotein (LDL) oxidation more effectively than probucol, a well-known commercially available antioxidant. In order to evaluate the anti-atherogenic potential of this medication, New Zealand White (NZW) rabbits were fed a normal diet for 12 weeks, a high cholesterol diet, a high cholesterol diet containing 1% probucol or a high cholesterol diet containing 5% water-soluble extract of GBH. Both GBH and probucol reduced plasma cholesterol levels. LDLs from the GBH-treated group were more resistant to Cu(2+)-induced oxidation and contained more vitamin E than LDLs from the high cholesterol diet group. Endothelial damage, determined at week 6, was reduced by 55% in the GBH group (P<0.01). GBH treatment reduced an atherosclerotic area in the abdominal aorta by 58% (P<0.05) and cholesterol deposition in the thoracic aorta by 55% (P<0.05). The severity of atherosclerosis in the GBH group was significantly reduced after an adjustment using cholesterol exposure as an index of the cholesterol-lowering effect. On the other hand, diet induced hyperlipidemic rabbits were given water extract of GBH in doses of 50 (Group B) and 200 mg/kg (Group C) and compared with controls (Group A). At 40 days after intervention in groups A, B and C, total and LDL cholesterol levels were significantly lowered (P<0.01). LDL/high density lipoprotein (HDL) ratio was also significantly decreased (P<0.01). This study concludes that the reduction in atherosclerosis by GBH relies not only on its cholesterol-lowering effect but also more heavily on its antioxidant potential, which prevents endothelial damage and inhibits LDL oxidative modification in hypercholesterolemic animals. PMID- 12757743 TI - Low-dose sulfur mustard primes oxidative function and induces apoptosis in human polymorphonuclear leukocytes. AB - Although considerable work has focused on understanding the processes of direct tissue injury mediated by the chemical warfare vesicant, sulfur mustard (2,2'-bis chloroethyl sulfide; SM), relatively little is known regarding the mechanisms of secondary injury caused potentially by the acute inflammatory response that follows SM exposure. Polymorphonuclear leukocytes (PMNs) play a central role in the initiation and propagation of inflammatory responses that, in some cases, result in autoimmune tissue damage. The potential for PMN-derived tissue damage following SM exposure may, in part, account for the protracted progression of the injury before it resolves. The current study was undertaken to evaluate the priming, oxidative function, and viability of PMN following exposure to low doses of SM such as those that might remain in tissues as a result of topical exposure. Our results demonstrate that doses of SM ranging from 25 to 100 microM primed PMN for oxidative burst in response to activation by fMLP, and that doses of SM ranging from 50 to 100 microM induced PMN apoptosis. Understanding the mechanisms through which SM directly affects PMN activation and apoptosis will be of critical value in developing novel treatments for inflammatory tissue injury following SM exposure. PMID- 12757744 TI - Effects of tetracaine and bupivacaine on chemiluminescence generated by formyl methionyl-leucyl-phenylalanine-stimulated human leukocytes and cell-free systems. AB - We investigated the abilities of an ester-type local anesthetic tetracaine and an amide-type local anesthetic bupivacaine to inhibit reactive oxygen and/or nitrogen species generated by either human leukocytes or cell-free systems via luminol- and lucigenin-enhanced chemiluminescence (CL). Tetracaine (96+/-1%, n=6, 1 mM) and bupivacaine (97+/-0.4%, n=5, 1 mM) significantly inhibited FMLP-induced CL in leukocyte assay. In cell-free experiments, CL due to superoxide production was significantly inhibited by tetracaine (23+/-2%, n=6) and bupivacaine (25+/ 4%, n=6) at 1 mM. Although bupivacaine was ineffective on H(2)O(2)-induced CL, tetracaine activated H(2)O(2)-induced luminol CL. Additionally, tetracaine inhibited FeSO(4)-induced CL (42+/-2%, n=6, 1 mM). In hypochlorous acid (HOCl) induced CL assay, 70+/-10% (n=5) and 57+/-4% (n=15) inhibitions were observed by tetracaine and bupivacaine, respectively. Peroxynitrite-induced luminol (54+/-7%, n=7, tetracaine, and 26+/-5%, n=8, bupivacaine, at 1 mM) and lucigenin CL (58+/ 3%, n=6, tetracaine, and 22+/-14%, n=9, bupivacaine, at 1 mM) were markedly inhibited. Tetracaine interacted with superoxide, hydroxyl radical, HOCl and peroxynitrite, while bupivacaine scavenged superoxide, HOCl and peroxynitrite. These direct scavenging properties of these drugs might be involved in the inhibition observed in leukocyte free radical release. In general, a decrease in CL-response was seen with higher concentrations (0.1-1 mM) of the local anesthetics, it is likely that tetracine and bupivacaine at therapeutic concentrations do not suppress leukocyte function in vivo. PMID- 12757745 TI - Time course of acute-phase response induced by Tityus serrulatus venom and TsTX-I in mice. AB - Animal venom can induce systemic alterations similar to those observed in acute phase inflammatory response. In the present study, we report the systemic (circulatory) and local (peritoneal cavity) effects induced by Tityus serrulatus venom and its major toxin TsTX-I (Ts1) in mice over various time periods. Both the venom and TsTX-I elicited quite similar responses in most assays. Responses included reduction of albumin, increased C-reactive protein, IL-6, IL-1alpha and TNF-alpha. Local and systemic leucocytosis, with a predominance of polymorphonuclear cells, was also observed. These effects show that a systemic inflammation-like syndrome is triggered during the severe envenomation caused by the T. serrulatus sting. The initial increases of albumin and total protein were probably consequences of the dehydration that occurs at the beginning of envenomation. Time-course analysis of these effects shows that responses are most pronounced on the first day after poisoning. However, leucocytosis and changes in acute-phase protein concentrations can be observed up to 7 days after envenomation. PMID- 12757746 TI - Liver fluke (Fasciola hepatica)-derived peptides activate rat peritoneal mast cells. AB - Short peptides with sequences derived from those found in the tegumental antigen of Fasciola hepatica have been synthesised. Incubation of some of these peptides with rat peritoneal mast cells resulted in the degranulation of the cells as measured by a histamine release assay. This activity was shown to be associated with the proline-lysine-proline motif, which is responsible for the induction of mast cell degranulation by the mammalian bioactive peptide substance P. Studies on the mode of action of the fluke-derived peptide indicated that it was operating through the same biochemical pathways as substance P. The implications of these findings for the development of immune responses during parasite infections are discussed. PMID- 12757747 TI - Target-mediated control of neural differentiation. AB - The development of the nervous system entails the coordination of the spatial and chemical development of both pre- and postsynaptic elements. This coordination is accomplished by signals passing between neurons and the target cells that they innervate. This review focuses on well-characterized examples of target-mediated neuronal differentiation in the central and peripheral nervous systems. These include control of neurogenesis in the leech by male genitalia, presynaptic differentiation induced by postsynaptic molecules expressed by skeletal muscle, postsynaptic adhesion molecules that induce presynaptic differentiation in the central nervous system (CNS), target-mediated control of neurotransmitter phenotype in peripheral neurons, and target-regulated control of neuronal nicotinic acetylcholine receptors (nAChRs) and large conductance calcium activated potassium channels (BK). The detailed understanding of these processes will uncover signals critical for the directed differentiation of stem cells as well as identify future targets for therapies in neural regeneration that promote the reestablishment of functional connections. PMID- 12757748 TI - Diabetic neuropathy and nerve regeneration. AB - Diabetic neuropathy is the most common peripheral neuropathy in western countries. Although every effort has been made to clarify the pathogenic mechanism of diabetic neuropathy, thereby devising its ideal therapeutic drugs, neither convinced hypotheses nor unequivocally effective drugs have been established. In view of the pathologic basis for the treatment of diabetic neuropathy, it is important to enhance nerve regeneration as well as prevent nerve degeneration. Nerve regeneration or sprouting in diabetes may occur not only in the nerve trunk but also in the dermis and around dorsal root ganglion neurons, thereby being implicated in the generation of pain sensation. Thus, inadequate nerve regeneration unequivocally contributes to the pathophysiologic mechanism of diabetic neuropathy. In this context, the research on nerve regeneration in diabetes should be more accelerated. Indeed, nerve regenerative capacity has been shown to be decreased in diabetic patients as well as in diabetic animals. Disturbed nerve regeneration in diabetes has been ascribed at least in part to all or some of decreased levels of neurotrophic factors, decreased expression of their receptors, altered cellular signal pathways and/or abnormal expression of cell adhesion molecules, although the mechanisms of their changes remain almost unclear. In addition to their steady-state changes in diabetes, nerve injury induces injury-specific changes in individual neurotrophic factors, their receptors and their intracellular signal pathways, which are closely linked with altered neuronal function, varying from neuronal survival and neurite extension/nerve regeneration to apoptosis. Although it is essential to clarify those changes for understanding the mechanism of disturbed nerve regeneration in diabetes, very few data are now available. Rationally accepted replacement therapy with neurotrophic factors has not provided any success in treating diabetic neuropathy. Aside from adverse effects of those factors, more rigorous consideration for their delivery system may be needed for any possible success. Although conventional therapeutic drugs like aldose reductase (AR) inhibitors and vasodilators have been shown to enhance nerve regeneration, their efficacy should be strictly evaluated with respect to nerve regenerative capacity. For this purpose, especially clinically, skin biopsy, by which cutaneous nerve pathology including nerve regeneration can be morphometrically evaluated, might be a safe and useful examination. PMID- 12757749 TI - Intracellular sorting and transport of proteins. AB - The secretory and endocytic pathways of eukaryotic organelles consist of multiple compartments, each with a unique set of proteins and lipids. Specific transport mechanisms are required to direct molecules to defined locations and to ensure that the identity, and hence function, of individual compartments are maintained. The localisation of proteins to specific membranes is complex and involves multiple interactions. The recent dramatic advances in understanding the molecular mechanisms of membrane transport has been due to the application of a multi-disciplinary approach, integrating membrane biology, genetics, imaging, protein and lipid biochemistry and structural biology. The aim of this review is to summarise the general principles of protein sorting in the secretory and endocytic pathways and to highlight the dynamic nature of these processes. The molecular mechanisms involved in this transport along the secretory and endocytic pathways are discussed along with the signals responsible for targeting proteins to different intracellular locations. PMID- 12757750 TI - On the structure and function of apolipoproteins: more than a family of lipid binding proteins. AB - Exchangeable apolipoproteins have been the subject of intense biomedical investigation for decades. However, only in recent years the elucidation of the three-dimensional structure reported for several members of the apolipoprotein family has provided insights into their functions at a molecular level for the first time. Moreover, the role of exchangeable apolipoproteins in several cellular events distinct from lipid metabolism has recently been described. This review summarizes these contributions, which have not only allowed the identification of the apolipoprotein domains that determine substrate binding specificity and/or affinity but also the plausible molecular mechanism(s) involved. PMID- 12757751 TI - Muscle satellite cells. AB - Skeletal muscle satellite cells are quiescent mononucleated myogenic cells, located between the sarcolemma and basement membrane of terminally-differentiated muscle fibres. These are normally quiescent in adult muscle, but act as a reserve population of cells, able to proliferate in response to injury and give rise to regenerated muscle and to more satellite cells. The recent discovery of a number of markers expressed by satellite cells has provided evidence that satellite cells, which had long been presumed to be a homogeneous population of muscle stem cells, may not be equivalent. It is possible that a sub-population of satellite cells may be derived from a more primitive stem cell. Satellite cell-derived muscle precursor cells may be used to repair and regenerate damaged or myopathic skeletal muscle, or to act as vectors for gene therapy. CELL FACTS: (1) Number of cells in body: 2 x 10(7) to 3 x 10(7) myonuclei/g, 20-25 kg muscle in average man; 2 x 10(5) to 10 x 10(5) satellite cells/g, i.e. approximately 1 x 10(10) to 2 x 10(10) satellite cells per person. (2) Main functions: repair and maintenance of skeletal muscle. (3) Turnover rate: close to zero in non-traumatic conditions high in disease or severe trauma. PMID- 12757752 TI - Kallmann's syndrome: molecular pathogenesis. AB - Kallmann's syndrome (KS) is a genetic condition characterised by hypogonadotrophic hypogonadism (HH) and anosmia; although these are the defining features of the condition, additional neurological and non-neurological sequel may also occur depending on the specific mode of inheritance. KS affects about 1 in 8000 males and 1 in 40,000 females, with most presentations being of the 'sporadic' type. Of the inherited forms, hitherto, only the gene responsible for the X-linked form (X-KS), namely KAL-1, has been identified and the encoded protein, anosmin-1, consists primarily of a whey acidic protein (WAP) and fibronectin-like type III (FnIII) domains which appear to mediate distinctly different protein functions. The WAP/FnIII combination is conserved in anosmins across species and recent studies in rodents and in Caenorhabditis elegans demonstrate that anosmin functions in both axonal targeting and branching. Screening for loci that modify these phenotypes in C. elegans has identified heparan-6-O-sulphotransferase as a key interactor mediating anosmin-1 function. Furthermore, over-expression and loss of function of the C. elegans Kal-1 gene disrupt epidermal morphogenesis, resulting in ventral enclosure and male tail formation defects. These findings provide novel insights into the molecular pathogenesis of X-KS. PMID- 12757753 TI - Evaluation of the fluidity and functionality of the renal cortical brush border membrane in experimental diabetes in rats. AB - The present study was aimed at addressing the effect of hyperglycemia on the renal cortical brush border membrane. The fluidity and the functionality of the renal cortical brush border membrane have been evaluated after 6 weeks of streptozotocin-induced diabetes in rats. Lipid peroxidation and protein oxidation were first performed to confirm a state of oxidative stress. The fluidity of the brush border membrane of diabetic rats decreased significantly by 15.76%. There was an increase in the amount of early (19.39%) and advanced (42.23%) glycation end-products suggesting the accumulation of significant amount of non-enzymic glycation products at 6 weeks of diabetes. Although, the activities of both gamma glutamyl transpeptidase and alkaline phosphatase of the brush border membrane decreased, that of the latter decreased to a significant extent with an increase in K(m) (81%) and no change in the V(max). A study of the activities of glutathione-dependent antioxidant enzymes in the renal cortical homogenates showed that the activities of glutathione peroxidase and glyoxalase II were altered significantly. Our study seems to suggest that increased free radical generation accompanied by non-enzymic glycation may be responsible for oxidative stress and an increased rigidity of the diabetic brush border membrane. Alkaline phosphatase may thus serve as a potentially useful marker of free radical induced damage to the renal cortical brush border membrane. The results also suggest that enhanced susceptibility to oxidative stress during early stages may be an important factor in the development of secondary complications of diabetes. PMID- 12757754 TI - Glycoprotein Ib-IX-V. AB - Glycoprotein (GP) Ib-IX-V is a remarkable platelet adhesion receptor of the leucine-rich repeat family. It has evolved to fulfil its major function of initiating platelet aggregation (thrombus formation) at high-shear stress in flowing blood. In addition to binding von Willebrand factor (vWF) in subendothelial matrix or plasma to trigger platelet aggregation, GPIb-IX-V also binds counter-receptors, alphaMbeta2 (Mac-1) on neutrophils or P-selectin on activated platelets or endothelial cells. GPIb-IX-V ligands also include alpha thrombin, clotting factors XI/XIIa, and high-molecular-weight kininogen. Interactions involving GPIb-IX-V are therefore central to vascular processes of thrombosis and inflammation, and the receptor is under intense scrutiny as a potential therapeutic target. PMID- 12757755 TI - The molecular pathogenesis of hepatic encephalopathy. AB - Hepatic encephalopathy (HE) incorporates a spectrum of neuropsychiatric abnormalities seen in patients with liver dysfunction with a potential for full reversibility. Distinct syndromes are identified in acute liver failure and cirrhosis. Rapid deterioration in consciousness level and increased intracranial pressure that may result in brain herniation and death are a feature of acute liver failure whereas manifestations of HE in cirrhosis include psychomotor dysfunction, impaired memory, increased reaction time, sensory abnormalities, poor concentration and in severe forms, coma. For over a 100 years ammonia has been considered central to its pathogenesis. In the brain, the astrocyte is the main site for ammonia detoxification, during the conversion of glutamate to glutamine. An increased ammonia level raises the amount of glutamine within astrocytes, causing an osmotic imbalance resulting in cell swelling and ultimately brain oedema. The present review focuses upon the molecular mechanisms involved in the pathogenesis of HE. Therapy of HE is directed primarily at reducing ammonia generation and increasing its detoxification. PMID- 12757756 TI - The Gardos channel: a review of the Ca2+-activated K+ channel in human erythrocytes. AB - Ca(2+)-dependent K(+) efflux from human erythrocytes was first described in the 1950s. Subsequent studies revealed that a K(+)-specific membrane protein (the Gardos channel) was responsible for this phenomenon (the Gardos effect). In recent years several types of Ca-activated K(+) channel have been identified and studied in a wide range of cells, with the erythrocyte Gardos channel serving as both a model for a broader physiological perspective, and an intriguing component of erythrocyte function. The existence of this channel has raised a number of questions. For example, what is its role in the establishment and maintenance of ionic distribution across the red cell membrane? What role might it play in erythrocyte development? To what extent is it active in circulating erythrocytes? What are the cell-physiological implications of its dysfunction?This review summarises current knowledge of this membrane protein with respect to its function and structure, its physiological roles (some putative) and its contribution to various disease states, and it provides an introduction to adaptable NMR methods, which is our own area of technical expertise, for such ion transport analysis. PMID- 12757757 TI - Cell signaling and transcription factor activation by asbestos in lung injury and disease. AB - Signaling pathways initiated at the external cell surface or within the cytoplasm regulate transactivation of transcription factors and gene expression that are causally related to a number of critical cellular outcomes including proliferation, apoptosis, cell survival, and production of inflammatory cytokines. Asbestos, a ubiquitous pathogenic group of mineral fibers, can stimulate gene expression in a variety of cell types in the lung via intracellular signaling pathways. These cell signaling cascades may be initiated through receptor-mediated events or integrins. Alternatively, they may be stimulated by oxidants generated both during phagocytosis of minerals and/or by redox reactions on the mineral surface. Once initiated, these pathways can lead to promotion of gene expression critical to cellular injury, proliferation and inflammation-events leading to the development of fibroproliferative diseases of the lung and pleura. The elucidation and relevance of critical signaling cascades to lung injury or repair following asbestos exposure could aid in developing strategies to prevent or treat asbestos-associated lung and pleural diseases. PMID- 12757758 TI - Ternary complex factors: prime nuclear targets for mitogen-activated protein kinases. AB - Ternary complex factors (TCFs), a subgroup of the ETS protein family, were first described in the context of c-fos gene regulation. Subsequently, their early identification as nuclear targets for mitogen-activated protein kinases served to exemplify the fundamental links in eukaryotic cells between growth factor mediated signalling pathways and gene control. This article provides an overview of recent work on ternary complex factors, addressing their expression and molecular structure, as well as how selective interactions with members of other protein families serve to up-1 regulate or restrict their activity. Although only one genetic study on ternary complex factors has been published to date, unravelling of the underlying molecular events provides a basis for tentative predictions about their biological roles in mammalian organisms. PMID- 12757759 TI - Unfolding and inactivation of Ampullarium crossean beta-glucosidase during denaturation by guanidine hydrochloride. AB - Changes of activity and conformation of Ampullarium crossean beta-glucosidase in different concentrations of guanidine hydrochloride (GuHCl) have been studied by measuring the fluorescence spectra and its relative activity after denaturation. The fluorescence intensity of the enzyme decreased distinctly with increasing guanidine concentrations, the emission peaks appeared red shifted (from 338.4 to 350.8 nm), whereas a new fluorescence emission peak appeared near 310 nm. Changes in the conformation and catalytic activity of the enzyme were compared. A corresponding rapid decrease in catalytic activity of the enzyme was also observed. The extent of inactivation was greater than that of conformational changes, indicating that the active site of the enzyme is more flexible than the whole enzyme molecule. k(+0)>k(+0)' also showed that the enzyme was protected by substrate to a certain extent during guanidine denaturation. PMID- 12757760 TI - Localization and hormonal control of serine dehydratase during metabolic acidosis differ markedly from those of phosphoenolpyruvate carboxykinase in rat kidney. AB - Serine dehydratase (SDH) is abundant in the rat liver but scarce in the kidney. When administrated with dexamethasone, the renal SDH activity was augmented 20 fold, whereas the hepatic SDH activity was affected little. In situ hybridization and immunohistochemistry revealed that SDH was localized to the proximal straight tubule of the nephron. To address the role of this hormone, rats were made acidotic by gavage of NH(4)Cl. Twenty-two hours later, the SDH activity was increased three-fold along with a six-fold increment in the phosphoenolpyruvate carboxykinase (PEPCK) activity, a rate-limiting enzyme of gluconeogenesis. PEPCK, which is localized to the proximal tubules under the normal condition, spreads throughout the entire cortex to the outer medullary rays by acidosis, whereas SDH does not change regardless of treatment with dexamethasone or NH(4)Cl. When NH(4)Cl was given to adrenalectomized rats, in contrast to the SDH activity no longer increasing, the PEPCK activity responded to acidosis to the same extent as in the intact rats. A simultaneous administration of dexamethasone and NH(4)Cl into the adrenalectomized rats fully restored the SDH activity, demonstrating that the rise in the SDH activity during acidosis is primarily controlled by glucocorticoids. The present findings clearly indicate that the localization of SDH and its hormonal regulation during acidosis are strikingly different from those of PEPCK. PMID- 12757762 TI - Q9, a content-balancing accuracy index to evaluate algorithms of protein secondary structure prediction. AB - A content-balancing accuracy index, called Q(9), has been proposed to evaluate algorithms of protein secondary structure prediction. Here the content-balancing means that the evaluation is independent of the contents of helix, strand and coil in the protein being predicted. It is shown that Q(9) is much superior to the widely used index Q(3). Therefore, algorithms are more objectively evaluated by Q(9) than Q(3). Based on 396 non-homologous proteins, five algorithms of secondary structure prediction were evaluated and compared by the new index Q(9). Of the five algorithms, PHD turned out to be the unique algorithm with an average Q(9) better than 60%. Based on the new index, it is shown that the performance of the consensus method based on a jury-decision from several algorithms is even worse than that of the best individual method. Rather than Q(3), we believe that Q(9) should be used to evaluate algorithms of protein secondary structure prediction in future studies in order to improve prediction quality. PMID- 12757761 TI - A simple method of rat renal brush border membrane preparation using polyethylene glycol precipitation. AB - A simple method for preparation of brush border membranes (BBM) from rat kidney using polyethylene glycol (PEG) precipitation has been described. This method avoids the use of cations for the preparation, which might alter membrane lipid composition. These preparations were assessed for enrichment of marker enzymes, contamination by subcellular structures, lipid composition and transport function. An enrichment of 11.8910-fold of alkaline phosphatase, 13.9500-fold of amino peptidase and 13.6500-fold of gamma-glutamyl transpeptidase and an approximate yield of 60% were seen in the final membrane preparation as compared to the homogenate. There was very little contamination of basolateral membranes, peroxisomes, microsomes or lysosomes in the final membrane preparation. Analysis of sugars indicated high content of fucose and sailic acid as compared to hexoses. Isolated membranes appeared as vesicles as seen by electron microscopy. Lipid analysis indicated the presence of various neutral and phospholipids with a high content of sphingomyelin along with a cholesterol/phospholipid ratio of 0.4850. The isolated membrane vesicles were able to transport glucose. This study has shown a simple method of renal brush border membrane preparation, which is comparatively pure and functionally active. PMID- 12757763 TI - 5-Aminolevulinic acid synthesis in epimastigotes of Trypanosoma cruzi. AB - BACKGROUND AND AIMS: Trypanosoma cruzi is the causative agent of Chagas disease or American trypanosomiasis. The parasite manifests a nutritional requirement for heme compounds because of its biosynthesis deficiency. The aim of this study has been to investigate the presence of metabolites and enzymes of porphyrin pathway, as well as ALA formation in epimastigotes of T. cruzi, Tulahuen strain, Tul 2 stock. METHODS: Succinyl CoA synthetase, 5-aminolevulinic acid (ALA) synthetase, 4,5-dioxovaleric (DOVA) transaminase, ALA dehydratase and porphobilinogenase activities, as well as ALA, porphobilinogen (PBG), free porphyrins and heme content were measured in a parasite cells-free extract. Extracellular content of these metabolites was also determined. RESULTS: DOVA, PBG, porphyrins and heme were not detected in acellular extracts of T. cruzi. However ALA was detected both intra- and extracellularly This is the first time that the presence of ALA (98% of intracellularly formed ALA) is demonstrated in the extracellular medium of a parasite culture. Regarding the ALA synthesizing enzymes, DOVA transaminase levels found were low (7.13+/-0.49EU/mg protein), whilst ALA synthetase (ALA-S) activity was undetectable. A compound of non-protein nature, low molecular weight, heat unstable, inhibiting bacterial ALA-S activity was detected in an acellular extract of T. cruzi. This inhibitor could not be identified with either ALA, DOVA or heme. CONCLUSIONS: ALA synthesis is functional in the parasite and it would be regulated by the heme levels, both directly and through the inhibitor factor detected. ALA formed can not be metabolized further, because the necessary enzymes are not active, therefore it should be excreted to avoid intracellular cytotoxicity. PMID- 12757764 TI - The human hyaluronan synthase genes: genomic structures, proximal promoters and polymorphic microsatellite markers. AB - The glycosaminoglycan (GAG) hyaluronan (HA) is a key component of the vertebrate extracellular matrix (ECM) and is synthesised by the HA synthase (HAS) enzymes HAS1, HAS2 and HAS3 at the plasma membrane. Accumulating evidence emphasises the relevance of HA metabolism in an increasing number of processes of clinical interest including renal fibrosis and peritoneal mesothelial wound healing. In the present study, the genomic sequences and organisation of the genes encoding the human HAS isoforms were deduced, in silico, from reference cDNA and genomic sequence data. These data were confirmed in vitro by sequencing of PCR-amplified HAS exons and flanking genomic sequences, comparison with sequence data for the corresponding murine Has orthologues, rapid amplification of 5' cDNA ends analysis and luciferase reporter assays on putative proximal promoter sequences. The HAS1 gene comprised five exons, with the translation start site situated 9bp from the 3' end of exon 1. In contrast, the genomic structures for HAS2 and both HAS3 variants spanned four exons, exon 1 forming a discrete 5'-untranslated region (5'-UTR) and the translation start site lying at nucleotide 1 of exon 2. Dinucleotide microsatellite loci were identified in intron 1 of HAS1 and HAS2, and immediately upstream of the HAS3 gene and their utility as linkage markers demonstrated in genomic DNA (gDNA) studies. We thus present a comprehensive resource for mutation detection screening of all HAS exons and/or linkage analysis of each HAS gene in a variety of disorders for which they are attractive candidates. PMID- 12757765 TI - Cell adhesion-related gene expression by Helicobacter pylori in gastric epithelial AGS cells. AB - Helicobacter pylori (H. pylori) infection leads to gastroduodenal inflammation, peptic ulceration and gastric carcinoma. H. pylori may induce disease-specific gene expression in gastric epithelial cells. cDNA microarray for 352 cancer related genes was used to identify the genes altered by H. pylori (cagA positive) in gastric epithelial AGS cells. Expressions of the genes identified on the microarray and other genes closely associated with these genes were determined by reverse transcriptase-polymerase chain reaction (RT-PCR). Western blot analysis and cell adhesion assay were performed to confirm the protein levels of the genes and the role of the genes on cell adhesion in H. pylori-infected AGS cells. As a result, the expression of four genes (galectin 1, aldolase A, integrin alpha5, LIM domain only 7 (LMO7)) were up-regulated by H. pylori in AGS cells, determined by cDNA microarray. RT-PCR analysis showed that the genes up-regulated by H. pylori were the genes regulating cell-cell adhesion and cell-extracellular matrix interaction, such as galectin-1 and galectin-3, integrin alpha5, and LIM domain only 7 (LMO7), and cancer-related glycolytic enzyme aldolase A and C. Cell adhesion to extracellular matrix proteins such as poly-L-lysine and fibronectin was mediated by H. pylori-induced expression of integrin alpha5. RT-PCR and Western blot analysis showed that E-cadherin, regulating cell adhesion and contact cell inhibition, was decreased by H. pylori in AGS cells. In conclusion, the increased expression of cell adhesion molecules and decrease in E-cadherin expression by H. pylori might contribute to cell adhesion, invasion and possibly cell proliferation in gastric epithelial cells. PMID- 12757768 TI - Oral anticoagulants in patients with coronary artery disease: an inexpensive and effective strategy. PMID- 12757769 TI - Oral anticoagulation after myocardial infarction. AB - In the present review, the role of oral anticoagulants (OAC) in the secondary prophylaxis a long term after myocardial infarction (MI) is discussed in the light of the results from recently published large randomized controlled trials (RCTs). In particular, comparision with aspirin, alone or in combination with OAC, is presented. The pathophysiological role of atherothrombosis in MI, with special stress on the crucial role of thrombin generation representing a special rationale for OAC prophylaxis, is emphasized. Recent RCTs clearly show the superiority of OAC over aspirin to reduce clinical end points a long term after MI. A prerequisite for optimal effect of OAC is a knowledge of the narrow therapeutic window towards bleeding complications. This necessitates good patient compliance and strict control of the treatment. The INR threshold for effective prophylaxis with OAC after MI, when given alone or in combination with aspirin, is also presented. The possibility of self-management of INR controls with satisfactory quality and cost reduction is also highlighted. Finally, the combination of OAC with a target INR of 2.0-2.5 (2.3) and aspirin of 75 mg/day is especially recommended. PMID- 12757770 TI - Prevalence of factor V Leiden, FII G20210A, FXIII Val34Leu and MTHFR C677T polymorphisms in cancer patients with and without venous thrombosis. AB - Venous thromboembolism (VTE) is a common complication in patients with malignant disease. In addition to well-established acquired risk factors for VTE, several genetic risk factors, mainly related to the haemostatic system, are known to influence thrombotic risk. However, the contribution of gene abnormalities to thrombotic tendency in cancer patients remains poorly explored. We performed a prospective study to evaluate the prevalence and clinical significance of four gene variations (factor V Leiden [FVL], factor II G20210A, factor XIII Val34Leu and MTHFR C677T) in cancer patients, with and without VTE. Enrolled were 211 unrelated and unselected patients (M/F ratio 0.5, mean age 57 years, range 12-91 years) with a diagnosis of cancer, admitted to two University Oncology Clinics in the city of Sao Paulo, Southeastern Brazil. After admission, all patients were evaluated for the presence of symptoms and signs of VTE. Sixty-four patients (30.3%) had an episode of deep venous thrombosis (DVT) or pulmonary embolism (PE), which has been objectively verified; 147 patients (69.7%) had no evidence of VTE. FVL was found with a frequency of 1.5% and 2.7% in the VTE and non-VTE group, respectively (odds ratio [OR] for VTE 0.6, 95% CI: 0.06-5.3). FII G20210A was found in 1.5% and 1.3% of thrombotic and nonthrombotic patients, respectively, yielding an OR of 1.2 (95% CI: 0.1-13.1). FXIII Val34Leu was detected in 29.6% of the thrombotic patients and in 28.5% of the non-thrombotic patients (OR 1.1, 95% CI: 0.5-2). MTHFR 677T was present in 53.1% and 60.5% of patients with and without thrombosis, respectively (OR 0.8, 95% CI: 0.4-1.4). The present data do not point to an association between the four polymorphisms here investigated and the risk of VTE in cancer patients. PMID- 12757771 TI - Endothelial cells release phenotypically and quantitatively distinct microparticles in activation and apoptosis. AB - BACKGROUND: Endothelial cells (EC) shed endothelial microparticles (EMP) in activation and apoptosis. OBJECTIVES: We compared the antigenic expression of EMP species released during activation as compared to apoptosis, in three cell lines. METHODS: EC from renal and brain microvascular (MiVEC) and coronary macrovascular (MaVEC) origin were incubated with TNF-alpha to induce activation, or deprived of growth factors to induce apoptosis. Antigens expressed on EMP and EC were assayed flow cytometrically and included constitutive markers (CD31, CD51/61, CD105), inducible markers (CD54, CD62E and CD106), and annexin V binding. RESULTS: It was found that in apoptosis, constitutive markers in EMP were markedly increased (CD31>CD105), with a concomitant decrease in expression in EC. Annexin V EC surface binding and annexin V+ EMP were more sharply increased in apoptosis than in activation. In contrast, in activation, inducible markers in EMP were markedly increased in both EMP and EC (CD62E>CD54>CD106). Coronary MaVEC released significantly less EMP than MiVEC. CONCLUSION: EC release qualitatively and quantitatively distinct EMP during activation compared to apoptosis. Analysis of EMP phenotypic signatures may provide clinically useful information on the status of the endothelium. PMID- 12757772 TI - Aprotinin restores the adhesive capacity of dysfunctional platelets. AB - The post-operative coagulopathy associated with cardiopulmonary bypass (CPB) is known to be predominantly related to platelet dysfunction. The use of the serine protease inhibitor aprotinin dramatically reduces CPB associated hemorrhage and is thought to act primarily through the inhibition of plasmin without directly influencing platelets. Our data indicate that there is a direct effect of aprotinin on platelet adhesion, which has not been previously reported. We found that when aprotinin was added to blood samples with poorly adhesive platelets, platelet adhesion significantly increased as measured by the percent coverage of denuded arterial segments in the Baumgartner perfusion chamber. In preliminary experiments using expired platelet concentrates or fresh whole blood, the addition of aprotinin induced a positive increase of 22+/-7.5 and 14+/-6.2 percentage point in platelet adhesion, respectively. A simulated CPB model that recirculated a unit of anticoagulated whole blood for 2 h was used (n=14) to induce a platelet adhesion defect similar to that seen in clinical CPB. At initiation of recirculation, platelet adhesion was 55+/-9.5% but dropped to 13+6.5% coverage after 2 h simulated CPB. The addition of aprotinin to the post recirculation samples induced a significant restoration of platelet adhesion back to 38+/-11% coverage. When epsilon amino-caproic acid with soybean trypsin inhibitor was added to post recirculation samples, there was no similar effect on adhesion scores. To compare these findings with surgical CPB, we collected one blood sample at the beginning and two at the end of CPB from each of seven open heart patients. Aprotinin was added to one of each of the post-CPB samples. Platelet adhesion at the onset of surgical CPB was only 39+/-11% in this patient group but dropped to 7+/-7% by the end. Similar to the model, the addition of aprotinin post-CPB restored adhesion to 29+/-11%. These results suggest some action of aprotinin other than its antiplasmin effect, and that platelet adhesion in general can be promoted by aprotinin. PMID- 12757773 TI - Enzymes that hydrolyze adenine nucleotides in diabetes and associated pathologies. AB - The activities of the enzymes NTPDase (E.C. 3.6.1.5, apyrase, ATP diphosphohydrolase, ecto-CD39) and 5'-nucleotidase (E.C. 3.1.3.5, CD73) were analyzed in platelets of type 2 diabetic, hypertensive and type 2 diabetic/hypertensive patients. The results showed an increase in platelet NTPDase activity in type 2 diabetic (34% and 72%), hypertensive (32% and 70%) and type 2 diabetic/hypertensive patients (30% and 55%) when compared to control (P<.01) with ATP and ADP as substrate, respectively. 5'-Nucleotidase activity was elevated in the hypertensive (60%) and type 2 diabetic/hypertensive (53%) groups when compared to the control and type 2 diabetic group (P<.01). No differences in sensitivity to inhibitors was detected between the platelets of controls and type 2 diabetic/hypertensive patients. No effects on the enzyme activities were observed when pharmacological doses of propranolol, captopril, furosemide, chlorpropamide, acetylsalicylic acid and glibenclamide were administered. Furthermore, changes in platelet adhesiveness and reactivity were found in all groups tested. In conclusion, we may postulate that NTPDase and 5'-nucleotidase from platelets are altered in patients with type 2 diabetes and hypertension. Probably, such alterations are involved in compensatory physiological responses in these diseases and are related to other important mechanisms of thromboregulation. PMID- 12757774 TI - Testosterone and cocaine: vascular toxicity of their concomitant abuse. AB - Over the last few years, several studies have described an increase in the use of anabolic-androgenic steroids (AAS). More important, frequency of AAS use was significantly associated with frequency of psychotropic drug use, such as cocaine. Since information is not available on the effects of their concomitant abuse, and taking into account that cocaine and testosterone, when singly abused, are known to induce severe adverse effects on vascular system, our purpose was to evaluate in vitro the combined effect of these drugs on platelet and endothelial functions. Results show that testosterone, at concentrations not exerting any appreciably acute effects on their own, is capable of potentiating the cocaine effect on endothelial and platelet functions, indicating that concomitant use of testosterone and cocaine could result in enhancement of the thrombotic risk ascribed to these drugs. PMID- 12757775 TI - Soluble P selectin in systemic sclerosis: relationship with von Willebrand factor, autoantibodies and diffuse or localised/limited disease. PMID- 12757776 TI - Characterization and functional analysis of TFPI-2 gene promoter in a human choriocarcinoma cell line. AB - Tissue factor pathway Inhibitor-2 (TFPI-2) is associated with extracellular matrices and plays a major role in cell migration and tumor invasion. In this study, a 4.8-kb human TFPI-2 gene 5'-flanking region was isolated, cloned and sequenced. Promoter region analysis revealed a high GC-rich content without canonical TATA and CAAT boxes but three transcription initiation sites were identified. Moreover, several putative binding sites for transcription factors were identified (MyoD, LYF1, NF-Y, GATA, oct-1, AP-1, Sp1, NF1, NF-kappa B and egr-1). To characterize potential regulatory regions, TFPI-2/luciferase promoter constructs were then transfected in human choriocarcinoma JEG-3 cells. We first showed that the minimal TFPI-2 promoter is located between -166 and -111 from the translation start site. Luciferase activity consistently increased after stimulation of JEG-3 cells by phorbol 12-myristate 13-acetate indicating that NF1, NF-kappa B and egr-1/Sp1 binding sites are crucial in inducible TFPI-2 expression. Moreover, negative regulatory regions included AP-1 binding sites were identified. This study demonstrates that the TFPI-2 gene promoter exhibits typical features of a housekeeping gene. PMID- 12757777 TI - Recombinant factor VIIa reduces bleeding in severely thrombocytopenic rabbits. AB - Severe thrombocytopenia is a common complication to intensive chemotherapeutic regimens. For bleeding episodes associated with severe thrombocytopenia, the current standard treatment is platelet transfusion. However, due to several transfusion complications such as transfusion-transmitted diseases, platelet refractoriness and immunomodulation, as well as increasing problems with sufficient supply of platelet products, it is imperative to search for alternatives to platelet transfusion. To test the efficacy of recombinant activated human coagulation factor VII (rFVIIa, NovoSeven) in thrombocytopenia, a preclinical study was conducted in thrombocytopenic rabbits. Thrombocytopenia was induced by a combination of gamma-irradiation and the use of platelet antibodies, and the effect of rFVIIa on nail cuticle bleeding was determined. Administration of rFVIIa at 2 mg/kg significantly shortened the prolonged bleeding time in thrombocytopenic animals (rFVIIa vs. control, median 23 min 41 s vs. 60 min, p=0.016) as well as significantly reducing the blood loss (rFVIIa vs. control, median: 8.8 vs. 12.2 nmol hemoglobin/ml, p=0.016). This effect was also reflected by a significant reduction of the prothrombin time, activated partial thromboplastin time, as well as improvement in clotting parameters in an in vitro thromboelastography thrombocytopenia model. Histopathological evaluation of kidney biopsies for the presence of micro thrombi did not reveal evidence of prothrombotic effects of rFVIIa in this model. These data demonstrate the haemostatic efficacy of rFVIIa in a rabbit model of severe thrombocytopenia. Clinical trials will be needed to further explore the potential of NovoSeven as a haemostatic agent in thrombocytopenic patients. PMID- 12757778 TI - Cloning and expression of rat coagulation factor VII. AB - Smaller and widely available animals such as rats are commonly used to evaluate antithrombotic drug candidates in vivo. However, the isolation and purification of FVII from rats and other species is very challenging because they are present in extremely low levels in plasma (approximately 10 nM). Furthermore, purification of FVII from other coagulation factors present in the plasma such as prothrombin, factor IX and factor X can often be very challenging and labor intensive. To facilitate studies on the role of the extrinsic pathway of coagulation in rats, a full-length cDNA-encoding rat factor VII was isolated using polymerase-mediated DNA amplification using a rat liver cDNA library. The cDNA codes for a 41-residue signal/propeptide region, followed by a 405-residue mature protein consisting of the light chain with gamma-carboxy glutamic acid (gla) including epidermal growth factor domains (EGF) and the heavy chain with the serine protease catalytic domain. Rat factor VII cDNA was transfected into human embryonic kidney 293 cells and several cell lines that constitutively express rat factor VII were established. The media from the stable lines expressing recombinant rat FVII were rapidly screened for functional activity and were found to normalize clotting time of FVII-depleted human plasma. The supernatants were also functionally active in the presence of tissue factor in chromogenic assays by measuring FVIIa activation using a tripeptide chromogenic substrate and in a two-stage, coupled assay measuring the generation of FXa. Recombinant rat FVII may be an important new tool in the development of novel antithrombotic drugs. PMID- 12757779 TI - Conifer needles as passive biomonitors of the spatial and temporal distribution of DDT from a point source. AB - Needles of two conifer species, Picea abies and Pinus nigra, were used as passive samplers for monitoring air contamination by sampling at increasing distances from a suspected point source of DDT. Needle concentrations declined with increasing distance downwind of the point source allowing to identify spatial and temporal trends of accumulation. This suggested that conifer needles are effective biomonitors of contamination levels in areas characterized by the presence of semi-volatile substances. Differences in uptake were apparent between the species. Needle morphology and structure were studied with scanning electron microscope (SEM) as were dimensional parameters (surface area, volume). The results suggest that the concentrations depend on a mechanism involving the inner structure of the needles, specifically the number and accessibility of resin channels rather than their surface area. Pine needles have more channels with greater accessibility than spruce. The results suggest that spruce is more suitable for short term measurement while pine for determining long term cumulative exposure. PMID- 12757780 TI - Effect of glyphosate on the microbial activity of two Brazilian soils. AB - Glyphosate [N-(phosphonomethyl)-glycine] is a broad-spectrum, non-selective, post emergence herbicide that is widely used in agricultural. We studied, in vitro, changes in the microbial activity of typical Hapludult and Hapludox Brazilian soils, with and without applied glyphosate. Glyphosate was applied at a rate of 2.16 mg glyphosate kg(-1) of soil and microbial activity was measured by soil respiration (evolution of CO(2)) and fluorescein diacetate (FDA) hydrolysis over a period of 32 days. We found an increase of 10-15% in the CO(2) evolved and a 9 19% increase in FDA hydrolyses in the presence of glyphosate compared with the same type of soil which had never received glyphosate. Soil which had been exposed to glyphosate for several years had the strongest response in microbial activity. Most probable number (MPN) counts showed that after 32 days incubation the number of actinomycetes and fungi had increased while the number of bacteria showed a slight reduction. After the incubation period, high pressure liquid chromatography (HPLC) detected the glyphosate metabolite aminomethyl phosphonic acid (AMPA), indicating glyphosate degradation by soil microorganisms. PMID- 12757781 TI - Carbonyl compounds in the urban environment of Athens, Greece. AB - The concentration levels of 15 selected carbonyl compounds in 62 samples were determined at two sites in Athens basin from June to December 2000. Formaldehyde was the most abundant species (0.05-39 microg m(-3)), which comprised from 22% to 37% of the total measured compounds, followed by acetaldehyde (4.32-49 microg m( 3)), acetone/acrolein (0.64-198 microg m(-3)) and butanal (0.79-140 microg m( 3)). The mean formaldehyde/acetaldehyde and acetaldehyde/propanal molar ratios were calculated. No significant seasonal differences were observed for all the carbonyls. Photochemical production was found to weigh upon atmospheric levels for 83-93% in summer days, dropping below 33% in the winter. The importance of formaldehyde and acetaldehyde as a source of hydroxyl radicals in Athens was also assessed. PMID- 12757782 TI - A high performance liquid chromatography method for determination of gas-phase hydrogen peroxide in ambient air using Fenton's chemistry. AB - A new method for determination of hydrogen peroxide in atmospheric samples is described. Cryogenically collected H2O2 is reacted with sodium salicylate in the presence of Fe(2+) to produce dihydroxybenzoate, which is separated from the reaction mixture by high performance liquid chromatography and detected by UV absorption. Measurements of atmospheric H2O2 were conducted in Las Vegas, NV from June 1999 to December 1999 to evaluate and characterize the method. Measured mixing ratios of H2O2 (there were also non-detects) ranged from 0.012 to 2.74 ppbv, with expected correlations to primary gaseous pollutants and strong seasonal variation consistent with a photochemically derived species. It was concluded that the method is easy to use and has sufficient sensitivity and selectivity to be useful in atmospheric monitoring. PMID- 12757783 TI - Evaluation of activated carbon fiber filter for sampling of organochlorine pesticides in environmental water samples. AB - A simple method for quantitative analyses of organic chlorine pesticides (OCPs) in environmental water samples such as rainwater, river water and seawater using activated carbon fiber filters (ACFF) is described. ACFF was used as adsorbent to collect the chemicals in water samples. The collection of OCPs was completed almost for one day by stirring the mixture of the sample and the ACFF chips at room temperature. The adsorbed OCPs on the ACFF could be extracted easily with toluene-ethanol (4:1) mixed solvent. The purified extract by a florisil column chromatograph was followed by the analysis using high-resolution gas chromatograph/high-resolution mass spectrometer. Recoveries of OCPs spiked to actual samples such as rainwater, river water and seawater samples were approximately more than 80%, and the coefficients of variations were within 10%. This method was applied to the actual samples and was confirmed to be applicable for monitoring sub-ng/l level OCPs in environmental water samples. PMID- 12757784 TI - Removal of mercury (II) from aqueous solution by activated carbon obtained from furfural. AB - The adsorption of Hg(II) from aqueous solution at 293 K by activated carbon obtained from furfural is studied. The carbon is prepared by polymerization of furfural following carbonization and activation of the obtained polymer material with water vapor at 800 degrees C. Adsorption studies of Hg(II) are carried out varying some conditions: treatment time, metal ion concentration, adsorbent amount and pH. It is determined that Hg(II) adsorption follows both Langmuir and Freundlich isotherms. The adsorption capacity of the carbon is 174 mg/g. It is determined that Hg(II) uptake increases with increasing pH. Desorption studies are performed with hot water. The percent recovery of Hg(II) is 6%. PMID- 12757785 TI - Photocatalytic production of hydrogen in single component and mixture systems of electron donors and monitoring adsorption of donors by in situ infrared spectroscopy. AB - The photocatalytic production of hydrogen using aqueous Pt/TiO2 suspension has been investigated in single component and mixture systems of electron donors (pollutants). The reaction systems consisted of oxalic acid, formic acid and formaldehyde, respectively. The adsorption of these donors on TiO2 was also monitored by in situ attenuated total reflection infrared spectroscopy (ATRIR). In the single component systems, the efficiency order of electron donors is as follows: H2C2O4 > HCOOH > HCHO. The order is consistent with the order of adsorption affinity of the electron donors on TiO2 determined by ATRIR, which suggests a link between the strength of surface interaction and the efficiency of photocatalytic hydrogen evolution. In the binary mixture systems, competitive inhibition kinetics is observed. When a donor adsorbed strongly on TiO2 in a state of saturated adsorption in a binary system, the overall rate of the hydrogen evolution is consistent with that of decomposition of the donor, and the system can be treated as a single component system. PMID- 12757786 TI - Photodegradation mechanism for bisphenol A at the TiO2/H2O interfaces. AB - Bisphenol A (BPA) can be decomposed photocatalytically under UV-illumination in aqueous TiO2 dispersion. The two methyl groups in BPA were initially attacked with .OH and/or .OOH radicals having strong oxidizing power, followed by the cleavage of the two phenyl moieties. Finally, the photomineralization to CO2 gas occurred via oxidative processes involving carboxylic acids and aldehydes. The decomposition of structurally similar substances of 4,4'-ethylidenebisphenol (EBP) and 4,4'-methylenebisphenol (MBP) was compared. The decomposition of BPA gave more kinds of intermediates such as 4-isopropylphenol, 4-ethylphenol, etc. On the other hand, that of EBP gave mainly 4-isopropylphenol and that of MBP gave a predominant product of 4-hydroxybenzaldehyde. These photooxidative pathways were proposed on the base of the evidence of oxidative intermediate formation. PMID- 12757787 TI - Biodegradation of potential diesel oxygenate additives: dibutyl maleate (DBM), and tripropylene glycol methyl ether (TGME). AB - The addition of oxygen-bearing compounds to diesel fuel considerably reduces particulate emissions. TGME and DBM have been identified as possible diesel additives based on their physicochemical characteristics and performance in engine tests. Although these compounds will reduce particulate emissions, their potential environmental impacts are unknown. As a means of characterizing their persistence in environmental media such as soil and groundwater, we conducted a series of biodegradation tests of DBM and TGME. Benzene and methyl tertiary butyl ether (MTBE) were also tested as reference compounds. Primary degradation of DBM fully occurred within 3 days, while TGME presented a lag phase of approximately 8 days and was not completely degraded by day 28. Benzene primary degradation occurred completely by day 3 and MTBE did not degrade at all. The total mineralized fractions of DBM and TGME achieved constant values as a function of time of approximately 65% and approximately 40%, respectively. Transport predictions show that, released to the environment, DBM and TGME would concentrate mostly in soils and waters with minimal impact to air. From an environmental standpoint, these results combined with the transport predictions indicate that DBM is a better choice than TGME as a diesel additive. PMID- 12757788 TI - Estimation of selected physicochemical properties for methylated naphthalene compounds. AB - Liquid aqueous solubility (S(w,L)), octanol/water partition coefficients (K(ow)), liquid vapor pressure (P(v,L)), and Henry's law constants (H(c)) were estimated for 20 methylated naphthalenes ranging from monomethyl to tetramethylnaphthalenes. Chromatographic methods were used for the estimation. Chromatographic retention measurements were conducted for 11 reference compounds and regressions were fit between the retention indices and the physicochemical properties. HPLC octadecylsilyl column with acetonitrile/water eluent was used for the estimation of S(w,L) and K(ow). Two GC columns, HP5-MS and a more hydrophobic HP-1, were tested for the estimation of P(v,L). Measured retention indices for the methylated naphthalenes were entered to the regression equations to calculate the physicochemical properties for these compounds. Literature values, where available, were used to validate the calculated values. The method accurately estimated the physicochemical properties. Estimated S(w,L) and P(v,L) decreased with the number of methyl groups. K(ow) increased with the number of methyl groups. There was no obvious relation between H(c) and the number of methyl groups. Log S(w,L) ranged from 0.885 for 1,2,5,6-tetramethylnaphthalene to 2.269 for 1-methylnaphthalene (mmol/m(3)). Log K(ow) varied from 3.89 for 1 methylnaphthalene to 4.95 for 1,2,5,6-tetramethylnaphthalene. Log P(v,L) ranged from -0.983 for 1,2,5,6-tetramethylnaphthalene to 0.789 for 2-methylnaphthalene (Pa). Log H(c) varied from 1.03 for 1,4,5-trimethylnaphthalene to 1.73 for 2,6 dimethylnaphthalene (Pa m(3)/mol). There were no apparent effects of GC column hydrophobicity on the accuracy of the results. Estimation of S(w,L) and K(ow) based on GC retention indices was not as accurate as with HPLC. Comparison of the estimated values with values predicted by EPIWIN indicated that EPIWIN is useful in giving order-of-magnitude prediction of physicochemical properties. PMID- 12757789 TI - Ionic composition of PM(10) in the area of Thessaloniki, Greece. AB - Airborne particulate matter (PM(10)) was collected from July 1997 to July 1998 at three locations in the city of Thessaloniki. PM(10) samples were analyzed for Cl( ), NO3(-), SO4(2-), Ca(2+), Mg(2+), Na(+), K(+) and NH4(+). The average PM(10) concentrations were found similar in all three sites with higher values in cold period. The ionic content comprised the 17-23% of the PM(10) mass and sulfate made up the 35-38% of the PM(10) ionic content with an average concentration of 4.80-7.26 microg m(-3). Good correlation was found for SO4(2-) and NO3(-) with Ca(2+), Mg(2+) and Cl(-). Two factors were found to influence the variance of ionic constituents in PM(10) by using factor analysis. Data evaluation considering wind direction showed that higher PM(10) and other ionic concentrations are associated with calm conditions, suggesting influences of local sources. PMID- 12757790 TI - Behavior of 1,3-dichloropropene and methyl isothiocyanate in undisturbed soil columns. AB - To develop alternatives to methyl bromide (MeBr) for soil disinfection under environmental Moroccan conditions, distribution and persistence of 1,3 dichloropropene (1,3-D) and methyl isothiocyanate (MITC) were tested in undisturbed soil columns (12 cm internal diameter, 1 m length). 1,3-D was injected at a 15 cm depth and directly followed by metam-sodium (a precursor of MITC), which was applied at the soil surface of the same column using a peristaltic pump. Concerning the distribution of these fumigants in the soil profiles, our results showed that 24 h after treatment, 1,3-D and MITC were concentrated at the 0-40 cm soil layers, and reached the deeper layers 48 h later. MITC and 1,3-D dissipation was studied and the half-life (DT50) measured were 6.5 and 8 days, respectively. Total volatilization losses reached 9% for MITC and 28% for 1,3-D. MITC and 1,3-D volatilization was found to be influence by soil water contents. The results show that by reducing volatilization, photodegradation and leaching of these fumigants a suitable alternative to MeBr use is offered. PMID- 12757791 TI - Contribution of dissolved sulfates and sulfites in hydrogen sulfide emission from stagnant water bodies in Sri Lanka. AB - Accumulation of sulfur-containing compounds and their bacterial mediated reductions have led to the emission of pungent odors from stagnant water bodies. This study is focused on the contribution of inorganic sulfur compounds in the emission of hydrogen sulfide. The measured dissolved oxygen levels have demonstrated good negative correlations with the dissolved sulfide levels implying the oxygen deficiency is the key for the reduction of sulfate ion and sulfite ion to sulfide ion. Particularly, the dissolved molar fractions of sulfide from the total dissolved sulfur compounds (sulfates, sulfites and sulfides) have a very good correlation with the dissolved oxygen for the stagnant water bodies except the artificially aerated prawn farms. For the stagnant water bodies with significant correlations, linear regressions are reported for them to be utilized in estimating one component of the regression from the measurement of the other. The measured data were further utilized to estimate the levels of hydrogen sulfide gas. The pH of the water bodies has confined much of the dissolved sulfides in the form of bisulfide ion and they can be easily escaped to the atmosphere upon acidification due to industrial discharges and/or acidic precipitations. The estimated levels of hydrogen sulfide just above the water surface were plotted for the most polluted stagnant water body in Sri Lanka for the pH range of 5-10 and temperature range of 25-35 degrees C. PMID- 12757792 TI - Removal of organic polyelectrolytes and their metal complexes by adsorption onto xonotlite. AB - Natural organic polyelectrolytes (humic and fulvic acids) and their metal complexes were removed by adsorption onto xonotlite. The removal percentages of humic and fulvic acids by xonotlite were approximately 80% and 30%, respectively. Humic acid removal from solution by adsorption onto xonotlite took place more readily than fulvic acid removal. The molecular weight distributions of the humic substances remaining in solution after adsorption with the xonotlite were measured with size exclusion chromatography. A comparison of molecular weight distributions demonstrated conclusively that large molecular weight components were adsorbed preferentially, indicating that adsorption efficiency depends on the number of functional groups of humic substances. Furthermore, the surface topography of the adsorbent was observed before and after adsorption by scanning electron microscopy. The calculated heat of adsorption was of 330 kJ mol(-1) which was evaluated from the Clapeyron-Clausius equation. Therefore, the adsorption type can be considered chemical. Since xonotlite can be easily synthesized and obtained at low cost, the adsorption method of humic and fulvic acids is superior to their precipitation. PMID- 12757793 TI - Studies on the sorption behaviors of nitrobenzene on marine sediments. AB - The sorption behaviors of nitrobenzene on marine sediments were systematically investigated in this study. The nitrobenzene sorption on both HCl-treated and untreated sediments accorded well with the linear sorption isotherm. It occurred primarily through partition function of organic carbon of sediments. In comparison, the sorption behavior of nitrobenzene on H2O2-treated sediments was nonlinear and conformed to Langmuir isotherm. Sorption of nitrobenzene on H2O2 treated sediment was mainly through surface function of sediment minerals such as clays. With the increase of ionic strength (salinity), solubility of nitrobenzene in solution would decrease. At the same time, the release of dissolvable part of organic carbon into water solution would also decrease. As a result, partition coefficient and saturate adsorption amount of nitrobenzene on marine sediments increased with increasing salinity of seawater. Contrary to the influence of salinity, partition coefficient and saturate adsorption amount of nitrobenzene decreased with increasing temperature. PMID- 12757794 TI - Behavior of methyl isothiocyanate in soils under field conditions in Morocco. AB - The behavior of methyl isothiocyanate (MITC), active metabolite of metam-sodium (MS), was studied under field conditions in Morocco. MS was applied through drip irrigation in: (i) uncovered soil, (ii) soil covered with transparent polyethylene, and (iii) soil covered with virtual impermeable film. Concentrations of MITC were determined at different soil depths to determine the distribution of MITC and the concentration-time product (CTP). Six hours after MS application, in a sandy soil, MITC reaches the 20-30 cm soil layer, but remains highly concentrated in the upper 10-20 cm soil layer. In a silty clay soil, MITC was concentrated in the upper 0-10 cm soil layer. The dissipation of MITC under different conditions of application was fast and complete after seven days. However, MITC dissipation time (DT(50)) was <24 h in sandy soil treated, but 63 h in silty clay soil. Under these application conditions of MS, the plastic film reduced MITC loss to the atmosphere but the plastic film quality did not affect the behavior of MITC. The use of plastic film maintained high MITC concentrations and appropriate CTP at different soil depths. PMID- 12757795 TI - Are data on light-induced ammonium release from dissolved organic matter consistent? PMID- 12757796 TI - On the sedimentary occurrence of chlorophyllone a. AB - The 13(2),17(3)-cyclopheophorbide a enol (CPP) is shown to convert mainly to a approximately 1:1 mixture of (13(2)R/S) chlorophyllones a (Chlone), when chromatographed over silica gel or alumina supports. 15(1) hydroxychlorophyllonelactone a and some other chlorophyll a related compounds are also tentatively identified as minor transformation products of CPP. This raises the possibility that the chlorophyllones reported in recent sediments may be analytical artifacts from CPP. However, data for the surface sediments from Lake Motte as well as literature data for other contemporary sediments show that, (i) they are not artifacts, (ii) considering that CPP is the intermediate compound in the formation of chlorophyllones from chlorophyll a, the hydroxylation of CPP in the sedimentary environment involves an enzymatic process leading preferentially to 13(2)S chlorophyllone a. PMID- 12757797 TI - Trabecular bone modulus-density relationships depend on anatomic site. AB - One outstanding issue regarding the relationship between elastic modulus and density for trabecular bone is whether the relationship depends on anatomic site. To address this, on-axis elastic moduli and apparent densities were measured for 142 specimens of human trabecular bone from the vertebra (n=61), proximal tibia (n=31), femoral greater trochanter (n=23), and femoral neck (n=27). Specimens were obtained from 61 cadavers (mean+/-SD age=67+/-15 years). Experimental protocols were used that minimized end-artifact errors and controlled for specimen orientation. Tissue moduli were computed for a subset of 18 specimens using high-resolution linear finite element analyses and also using two previously developed theoretical relationships (Bone 25 (1999) 481; J. Elasticity 53 (1999) 125). Resultant power law regressions between modulus and density did depend on anatomic site, as determined via an analysis of covariance. The inter site differences were among the leading coefficients (p<0.02), but not the exponents (p>0.08), which ranged 1.49-2.18. At a given density, specimens from the tibia had higher moduli than those from the vertebra (p=0.01) and femoral neck (p=0.002); those from the trochanter had higher moduli than the vertebra (p=0.02). These differences could be as large as almost 50%, and errors in predicted values of modulus increased by up to 65% when site-dependence was ignored. These results indicate that there is no universal modulus-density relationship for on-axis loading. Tissue moduli computed using methods that account for inter-site architectural variations did not differ across site (p>0.15), suggesting that the site-specificity in apparent modulus-density relationships may be attributed to differences in architecture. PMID- 12757798 TI - Muscle oxygen consumption, determined by NIRS, in relation to external force and EMG. AB - Local oxygen consumption in a muscle (VO(2)) can be determined by near infrared spectroscopy (NIRS). In principle it should be possible to use this measure to validate musculoskeletal models. However, the relationship between VO(2) and external force, or between VO(2) and surface EMG, as a measure for muscle activity, is hardly known. The aim of this study was: (1) to evaluate the characteristics of the relationship between VO(2) and external moments and (2) to determine whether differences exist between the EMG-moment relationship and the VO(2)-moment relationship. Subjects (n=5) were asked to perform isometric contractions exerting combinations of elbow flexion and pro/supination moments at force levels up to 70% of their maximum. Simultaneous surface-EMG and NIRS measurements were performed on the m. biceps breve (BB) and the m. brachioradialis (BR). A linear relationship was found between EMG and VO(2). For the BB VO(2) and EMG were linearly related to both the flexion moment and the pro/supination moment. However, for the BR only a linear relationship with flexion moment was found. As expected, based on the findings above, the relationship between VO(2) and elbow flexion moment can be described by a linear equation, under the conditions of this study (isometric, and force levels up to 70%). These findings suggest that load sharing is independent of force level and that next to EMG, VO(2) can be used for the validation of musculoskeletal models. PMID- 12757800 TI - Coping with perturbations to a layout somersault in tumbling. AB - Tumbling is a dynamic movement requiring control of the linear and angular momenta generated during the approach and takeoff phases. Both of these phases are subject to some variability even when the gymnast is trying to perform a given movement repeatedly. This paper used a simulation model of tumbling takeoff to establish how well gymnasts can cope with perturbations of the approach and takeoff phases. A five segment planar simulation model with torque generators at each joint was developed to simulate tumbling takeoffs. The model was customised to an elite gymnast by determining subject specific inertia and torque parameters and a simulation was produced which closely matched a performance of a layout somersault by the gymnast. The performance of a layout somersault was found to be sensitive to the approach characteristics and the activation timings but relatively insensitive to the elasticity of the track and maximum muscle strength. Appropriate variation of the activation timings used during the takeoff phase was capable of coping with moderate perturbations of the approach characteristics. A model of aerial movement established that variation of body configuration in the flight phase was capable of adjusting for takeoff perturbations that would lead to rotation errors of up to 8%. Providing the errors in perceiving approach characteristics are less than 5% or 5 degrees and the errors in timing activations are less than 7ms, perturbations in the approach can be accommodated using adjustments during takeoff and flight. PMID- 12757799 TI - An investigation of the interactions between lower-limb bone morphology, limb inertial properties and limb dynamics. AB - Bone mass and size clearly affect the safety and survival of wild animals as well as human beings, however, little is known about the interactions between bone size and movement dynamics. A modeling approach was used to investigate the hypothesis that increased bone cortical area causes increased limb moments of inertia, decreased lower-limb movement maximum velocities, and increased energy requirements to sustain submaximum lower-limb locomotion movements. Custom software and digital data of a human leg were used to simulate femur, tibia, and fibula cortical bone area increases of 0%, 22%, 50%, and 80%. Limb segment masses, center of mass locations, and moments of inertia in the sagittal plane were calculated for each bone condition. Movement simulations of unloaded running and cycling motions were performed. Linear regression analyses were used to determine the magnitude of the effect cortical area has on limb moment of inertia, velocity, and the internal work required to move the limbs at a given velocity. The thigh and shank moment of inertia increased linearly up to 1.5% and 6.9%, respectively for an 80% increase in cortical area resulting in 1.3% and 2.0% decreases in maximum unloaded cycling and running velocities, respectively, and in 3.0% and 2.9% increases in internal work for the cycling and running motions, respectively. These results support the hypothesis and though small changes in movement speed and energy demands were observed, such changes may have played an important role in animal survival as bones evolved and became less robust. PMID- 12757801 TI - Direct comparison of calculated hip joint contact forces with those measured using instrumented implants. An evaluation of a three-dimensional mathematical model of the lower limb. AB - Characterisation of hip joint contact forces is essential for the definition of hip joint prosthesis design requirements. In vivo hip joint contact force measurements have been made using instrumented hip joint prostheses. However, to allow determination of the range of values of joint contact force and their directions relative to anatomical structures in a range of subject groups sufficient to form an agreed data base it is necessary to adopt a different approach without the use of an implanted transducer. The use of mathematical models of the lower limb to examine the forces in soft tissues and at the joints has provided valuable insight into internal loading conditions. Several authors have proposed mathematical musculo-skeletal models. However, there have been only limited attempts at validation of these models. It is possible to use the results of in vivo force measurements from instrumented prostheses to validate the results calculated using the mathematical models. In this study two subjects with instrumented hip joint prostheses were studied. Forces at the hip joints were calculated using a three-dimensional model of the leg. Walking at slow, normal and fast speeds (0.97-2.01m/s), weight transfer from two to one leg and back again, and sit to stand were studied. Direct comparisons were made between the 'gold standard' measured hip joint contact forces and the calculated forces. There was general agreement between the calculated and measured forces in both pattern and magnitude. There were, however, discrepancies. Reasons for these differences in results are discussed and possible model developments suggested. PMID- 12757802 TI - Video-computer quantitative evaluation of thumb function using workspace of the thumb. AB - This study develops the basis for video-computer quantitative evaluation method for measuring degree of thumb impairment. The system evaluates the three dimensional space (workspace) within which the thumb-tip moves and computes its surface area. This study is intended to be the basis of a subsequent study which will compute the percentage of actual use compared to the "ideal" value for the given thumb length. Based on this quantitative measurement, the movement function of the impaired thumb can be identified objectively. Repeatability testing confirms that the video motion analysis system is a reliable tool for measuring the workspace of thumb-tip motion. It is also shown experimentally that there is a linear correlation between the surface area of thumb-tip motion workspace and the square of the thumb length. Accordingly, this method may be used to evaluate the functional abnormalities and deformities of the thumb. PMID- 12757803 TI - Analysis of creep strain during tensile fatigue of cortical bone. AB - During fatigue tests of cortical bone specimens, at the unload portion of the cycle (zero stress) non-zero strains occur and progressively accumulate as the test progresses. This non-zero strain is hypothesised to be mostly, if not entirely, describable as creep. This work examines the rate of accumulation of this strain and quantifies its stress dependency. A published relationship determined from creep tests of cortical bone (Journal of Biomechanics 21 (1988) 623) is combined with knowledge of the stress history during fatigue testing to derive an expression for the amount of creep strain in fatigue tests. Fatigue tests on 31 bone samples from four individuals showed strong correlations between creep strain rate and both stress and "normalised stress" (sigma/E) during tensile fatigue testing (0-T). Combined results were good (r(2)=0.78) and differences between the various individuals, in particular, vanished when effects were examined against normalised stress values. Constants of the regression showed equivalence to constants derived in creep tests. The universality of the results, with respect to four different individuals of both sexes, shows great promise for use in computational models of fatigue in bone structures. PMID- 12757804 TI - Modeling the bifurcating flow in an asymmetric human lung airway. AB - In a former paper, the inspiratory flow characteristics in a three-generation symmetric bifurcation airway have been numerically investigated using a control volume method to solve the fully three-dimensional laminar Navier-Stokes equations. The present paper extends the work to deal with asymmetric airway extracted from the 5th-11th branches of the model of Weibel (Morphometry of the Human Lung. New York Academic Press, Verlag, 1963) in order to more appropriately model human air passage. Computations are carried out in the Reynolds number range 200-1600, corresponding to mouth-air breathing rates ranging from 0.27 to 2.16l/s, representative for an averaged height man breathing from quiet to vigorous state. Particular attentions are paid to establishing relations between the Reynolds number and the overall flow characteristics, including flow patterns and pressure drop. The study shows that the ratios of airflow rate through the medial branches to that of their mother branches are the same, and this is also true for the ratios of airflow rate through the lateral branches. This partially explains why regular human breathing is not affected by airways of different sizes. PMID- 12757805 TI - Elastic properties of cancellous bone derived from finite element models of parameterized microstructure cells. AB - Evaluation of constitutive properties of cancellous bone and their relationships to microstructural parameters is a crucial issue in analysis of stresses and strains in bone tissues and simulation of their remodelling. Known limitations of experimental methods as well as of the micro-FE techniques make the analysis and homogenization of 'equivalent' trabecular microstructures an advantageous tool for this task. In this study, parameterized orthotropic constitutive models of cancellous bone are derived from finite element analysis of repeatable microstructure cells. Two cell types are analysed: cube- and prism-based. The models are fully three-dimensional, have realistic curvilinear shapes and are parameterized with three shape parameters. Variation of the parameters allows to imitate most of the typical microstructure patterns observed in real bones, along with variety of intermediate geometries. Finite element models of cells are generated by a special-purpose structured mesh generator for any arbitrary set of shape parameter values. Six static numerical tests are performed for an exhaustive number of parameter value sets (microstructure instances). Multi-point boundary conditions imposed on the models ensure mutual fitting of deformed neighbouring cells. Values of computed stresses allow to determine all coefficients of elastic orthotropic stiffness matrix. Results have a form of tabularized functions of elastic constants versus the shape parameters. Comparison of the results with micro-FE data obtained for a large set of cancellous bone specimens proves a good agreement, though evidently better in the case of the prism-based cell model. PMID- 12757806 TI - Microcrack accumulation at different intervals during fatigue testing of compact bone. AB - Fatigue damage in bone occurs in the form of microcracks. This microdamage contributes to the formation of stress fractures and acts as a stimulus for bone remodelling. A technique has been developed, which allows microcrack growth to be monitored during the course of a fatigue test by the application of a series of fluorescent chelating agents. Specimens were taken from bovine tibiae and fatigue tested in cyclic compression at a stress range of 80MPa. The specimens were stained before testing with alizarin and up to three other chelating agents were applied during testing to label microcracks formed at different times. Microcracks initiated in interstitial bone in the early part of a specimen's life. Further accumulation of microcracks is then suppressed until the period late in the specimen's life. Microcracks were found to be longer in the longitudinal than in the transverse direction. Only a small proportion of cracks are actively propagating; these are longer than non-propagating cracks. These results support the concept of a microstructural barrier effect existing in bone, whereby cracks initiate easily but slow down or stop at barriers such as cement lines. PMID- 12757807 TI - Rheology of concentrated suspensions of deformable elastic particles such as human erythrocytes. AB - New models for the viscosity of concentrated suspensions of deformable elastic particles are developed using the differential effective medium approach (DEMA). The models are capable of describing the rheological behavior of un-aggregated suspensions of human red blood cells (RBCs). With the increase in shear rate, a shear-thinning behavior is predicted similar to that observed in the case of un aggregated suspensions of RBCs. A decrease in relative viscosity and an enhancement of shear-thinning behavior is predicted when either the particle rigidity (elastic modulus) is decreased or the continuous medium viscosity is increased. These predictions are similar to those observed in suspensions of human RBCs. The proposed models are evaluated using experimental data on normal and hardened human RBC suspensions in protein-free saline. PMID- 12757808 TI - Experimental study of laminar blood flow through an artery treated by a stent implantation: characterisation of intra-stent wall shear stress. AB - The stimulation of endothelial cells by arterial wall shear stress (WSS) plays a central role in restenosis. The fluid-structure interaction between stent wire and blood flow alters the WSS, particularly between stent struts. We have designed an in vitro model of struts of an intra-vascular prosthesis to study blood flow through a 'stented' section. The experimental artery consisted of a transparent square section test vein, which reproduced the strut design (100x magnifying power). A programmable pump was used to maintain a steady blood flow. Particle image velocimetry method was used to measure the flow between and over the stent branches, and to quantify WSS. Several prosthesis patterns that were representative of the total stent strut geometry were studied in a greater detail. We obtained WSS values of between -1.5 and 1.5Pa in a weak SS area which provided a source of endothelial stimulation propitious to restenosis. We also compared two similar patterns located in two different flow areas (one at the entry of the stent and one further downstream). We only detected a slight difference between the weakest SS levels at these two sites. As the endothelial proliferation is greatly influenced by the SS, knowledge of the SS modification induced by the stent implantation could be of importance for intra-vascular prostheses design optimisation and thus can help to reduce the restenosis incidence rate. PMID- 12757809 TI - Bias compensated least squares estimate of the center of rotation. AB - It is shown that a recently published least squares method for the estimation of the average center of rotation is biased. Consequently, a correction term is proposed, and an iterative algorithm is derived for finding a bias compensated solution to the least squares problem.The accuracy of the proposed bias compensated least squares method is compared to the previously proposed least squares method by Monte-Carlo simulations. The tests show that the new method gives a substantial improvement in accuracy. PMID- 12757811 TI - Towards a realistic biomechanical model of the thumb: the choice of kinematic description may be more critical than the solution method or the variability/uncertainty of musculoskeletal parameters. AB - A biomechanical model of the thumb can help researchers and clinicians understand the clinical problem of how anatomical variability contributes to the variability of outcomes of surgeries to restore thumb function. We lack a realistic biomechanical model of the thumb because of the variability/uncertainty of musculoskeletal parameters, the multiple proposed kinematic descriptions and methods to solve the muscle redundancy problem, and the paucity of data to validate the model with in vivo coordination patterns and force output. We performed a multi-stage validation of a biomechanical computer model against our measurements of maximal static thumbtip force and fine-wire electromyograms (EMG) from 8 thumb muscles in each of five orthogonal directions in key and opposition pinch postures. A low-friction point-contact at the thumbtip ensured that subjects did not produce thumbtip torques during force production. The 3-D, 8 muscle biomechanical thumb model uses a 5-axis kinematic description with orthogonal and intersecting axes of rotation at the carpometacarpal and metacarpophalangeal joints. We represented the 50 musculoskeletal parameters of the model as stochastic variables based on experimental data, and ran Monte Carlo simulations in the "inverse" and "forward" directions for 5000 random instantiations of the model. Two inverse simulations (predicting the distribution of maximal static thumbtip forces and the muscle activations that maximized force) showed that: the model reproduces at most 50% of the 80 EMG distributions recorded (eight muscle excitations in 5 force directions in two postures); and well-directed thumbtip forces of adequate magnitude are predicted only if accompanied by unrealistically large thumbtip torques (0.64+/-0.28Nm). The forward simulation (which fed the experimental distributions of EMG through random instantiations of the model) resulted in misdirected thumbtip force vectors (within 74.3+/-24.5 degrees from the desired direction) accompanied by doubly large thumbtip torques (1.32+/-0.95Nm). Taken together, our results suggest that the variability and uncertainty of musculoskeletal parameters and the choice of solution method are not the likely reason for the unrealistic predictions obtained. Rather, the kinematic description of the thumb we used is not representative of the transformation of net joint torques into thumbtip forces/torques in the human thumb. Future efforts should focus on validating alternative kinematic descriptions of the thumb. PMID- 12757810 TI - Forefoot structural predictors of plantar pressures during walking in people with diabetes and peripheral neuropathy. AB - Various foot structures are thought to influence forefoot plantar pressures during walking. High peak plantar pressures (PPP) during walking in people with diabetes mellitus (DM) and peripheral neuropathy (PN) can cause skin breakdown. The question addressed by this study is "What are the primary forefoot structural factors that predict regional PPP during walking in groups of people with and without DM and PN?" Twenty people with DM and PN (mean age 55+/-9 years, 6 female, 14 male, BMI=33+/-8) and 20 people without DM, matched for gender, age, and BMI were tested. Measures of foot structure were taken from three-dimensional images constructed from spiral X-ray computed tomography. Peak plantar pressure data were recorded during walking. Hierarchical multiple regression analysis was used to predict regional PPP at the great toe and five metatarsal heads from selected structural and walking variables. Metatarsal phalangeal joint angle (hammer toe deformity) was the most important variable predicting pressure, accounting for 19-45% of the PPP variance at five of the six locations in the DM group. Soft tissue thickness, hallux valgus, and forefoot arthropathy were the most important predictors of PPP in the control group. Combinations of structural and walking variables accounted for 47-71% of the variance in the DM group and 52 83% of the variance of PPP during walking in the control group. These structural variables, especially hammer toe deformity, should be considered in attempts to develop strategies to reduce excessive forefoot PPP that may contribute to skin breakdown or other injury. PMID- 12757812 TI - Muscle mechanics and neuromuscular control. AB - The purpose of this paper is to demonstrate that the properties of the mechanical system, especially muscle elasticity and limb mass, to a large degree determine force output and movement. This makes the control demands of the central nervous system simpler and more robust. In human triceps surae, a muscle with short fibres and a long tendon, the time courses of the total muscle+tendon length and of the length of the contractile component (CC) alone in running are completely different. The muscle tendon complex shows first an eccentric phase with negative work, followed by a concentric phase. The CC, on the other hand, is concentric all the time. Moreover, the work that is done, is done at a speed that guarantees a high energetic efficiency. It is argued that this high efficiency is an in built property of the muscle mechanics for muscles with a compliant tendon and a low v(max). When a muscle, or a set of muscles, moves a mass, and the duration of the action is short with respect to the isometric time constant of the muscle, we may call it an 'elastic bounce contraction'. In such a case the mass-spring interaction largely determines the time course of the force, and the efficiency of muscle contraction is most of the time close to optimum. In a similar way, whole limbs can be modelled as springs, with a stiffness that can be modulated by flexing the joints more or less. The motor control task of the central nervous system is simple for such elastic bounce contractions: a block-like activation is sufficient, in which timing is critical, but activation level is not. It seems possible that a whole class of actions can be generated by an identical timing sequence, with only a modulation in activation amplitude. An example is walking or running at different speeds. PMID- 12757813 TI - A new method of estimating gauge length for porcine aortic valve test specimens. AB - Porcine aortic valve (PAV) cusps are folded and wrinkled in the in vitro state. In the tensile testing of PAV specimens, estimating gauge length (the length at which a specimen starts to offer measurable resistance to load) is often difficult and subjective. We have therefore developed a new method for estimating the gauge length of such tissues. The method is based on the observation that the specimen's gauge length can be associated with a stationary point on the slope of its load-length curve if loaded from a wrinkled state, or a state of slight compression. We represented the load-length response of test specimens in the low load, high-compliance region by a cubic function and determined the stationary point on the slope of the function using elementary calculus. The cubic function representation is fine-tuned by reducing or expanding an originally selected "test region" until the correlation coefficient of the cubic fit is maximized. The new method was applied to data obtained from the tensile testing of strips of heart valve tissue and was found to be objective, repeatable and robust. PMID- 12757815 TI - Spatial and temporal heterogeneity of superficial muscle strain during in situ fixed-end contractions. AB - Numerical models of contracting muscle offer a powerful tool to study local mechanical load. For validation of these models, the spatial and temporal distribution of strain was quantified in fixed-end contracting rat tibialis anterior muscle in situ at optimal muscle length (L(o)) and at 120 degrees plantar flexion as well as at 125 and 33Hz stimulation frequency. We studied the hypothesis that after termination of stimulation in situ muscle segments near the motor endplates elongate while segments away from the endplates shorten. We show that both spatial and temporal inhomogeneities in muscle deformation occurred during contraction. Muscle plateau shortening strain equalled 4.1%. Maximal plateau shortening of a muscle segment was much larger (9.6%) and occurred distally (at 0.26 of the scaled length of the muscle). Manipulating torque levels by decreasing the stimulation frequency at the same muscle length induced a decrease in torque ( approximately 20%) with a smaller effect on the level and no effect on the pattern of muscle deformation. During relaxation, distal segments actively shortened at the expense of proximal muscle segments, which elongated. The segments undergoing lengthening were nearer to motor endplates than segments undergoing shortening. In conclusion, the present study provides experimental data on magnitude of contraction-induced deformation needed for validation of numerical models. Local muscle deformation is heterogeneous both temporally and spatially and may be related to proximity to the motor endplates. PMID- 12757814 TI - Fall-related upper body injuries in the older adult: a review of the biomechanical issues. AB - Although the epidemiology of fall-related injuries is well established for the elderly population over 65 years of age, the biomechanics of how, when and why injuries do and do not occur when arresting a fall have received relatively little attention. This paper reviews the epidemiological literature in the MEDLINE data base pertinent to the biomechanics of fall-related injuries, including data on fall rates, fall-related injury rates, fall directions and types of injuries available. It also covers primary sources not listed on MEDLINE, along with the pertinent biomechanics literature. Many falls in older adults are in a forward direction, and as a result the upper extremities are one of the most commonly injured structures, presumably in protecting the head and torso. In this review emphasis is placed on what is, and what is not, known of the biomechanical factors that determine the impact forces and injury risk associated with upper extremity injuries in forward falls. While decreased bone mineral density may be contributory, it is not a reliable predictor of fracture risk. Evidence is presented that fall-related impact forces can be reduced by appropriate volitional arrest strategies. Further theoretical and experimental research is needed to identify appropriate fall-arrest strategies for the elderly, as well as the physical capacities and skills required to do so. Inexpensive interventions might then be developed to teach safe fall-arrest techniques to older individuals. PMID- 12757816 TI - Tracking the time course of word recognition with MEG. AB - Twenty years ago it was discovered that recognition of semantically unexpected words is associated with a special ERP signature - the N400. Pinpointing the precise functional significance of the N400 has, however, been difficult. Recent MEG studies of word processing reveal that, in fact, the N400 decomposes into several functionally distinct subcomponents, allowing for more fine-grained investigation of its significance. PMID- 12757817 TI - Chronometric explorations of disordered minds. AB - Patients with borderline personality disorder have unstable emotions, difficulty maintaining relationships and a high probability of self-inflicted damage. A recent experiment of Posner et al. that measured reaction-time differences derived from a simple task, revealed that alerting and orienting is normal in these patients. By contrast, and perhaps at the root of this personality disorder, the module responsible for cognitive conflict resolution is defective. PMID- 12757818 TI - How are qualia coupled to functions? AB - WHAT DETERMINES THE NATURE OF SUBJECTIVE EXPERIENCE: associated behavioural functions or mediating neural activity? A recent analysis by Hurley and Noe of a variety of cases of behavioural and neural plasticity shows that, under different conditions, either can predominate. This adds to other efforts to transfer the doctrine of functionalism from philosophical debate to empirical scrutiny, where it is hoped that it may eventually be resolved. PMID- 12757819 TI - The deferential brain in action: Response to Jeffrey Gray. PMID- 12757820 TI - Neural circuitry underlying pain modulation: expectation, hypnosis, placebo. AB - The ability to predict the likelihood of an aversive event is an important adaptive capacity. Certainty and uncertainty regarding pain cause different adaptive behavior, emotional states, attentional focus, and perceptual changes. Recent functional neuroimaging studies indicate that certain and uncertain expectation are mediated by different neural pathways-the former having been associated with activity in the rostral anterior cingulate cortex and posterior cerebellum, the latter with activation changes in the ventromedial prefrontal cortex, mid-cingulate cortex and hippocampus. Expectation plays an important role not only in its modulation of acute and chronic pain, but also in other disorders which are characterized by certain expectation (specific phobias) or uncertain expectation (generalized anxiety disorder) of aversive events. PMID- 12757821 TI - The representation of grammatical categories in the brain. AB - Language relies on the rule-based combination of words with different grammatical properties, such as nouns and verbs. Yet most research on the problem of word retrieval has focused on the production of concrete nouns, leaving open a crucial question: how is knowledge about different grammatical categories represented in the brain, and what components of the language production system make use of it? Drawing on evidence from neuropsychology, electrophysiology and neuroimaging, we argue that information about a word's grammatical category might be represented independently of its meaning at the levels of word form and morphological computation. PMID- 12757822 TI - Is perception discrete or continuous? AB - How does conscious perception evolve following stimulus presentation? The idea that perception relies on discrete processing epochs has been often considered, but never widely accepted. The alternative, a continuous translation of the external world into explicit perception, although more intuitive and subjectively appealing, cannot satisfactorily account for a large body of psychophysical data. Cortical and thalamocortical oscillations in different frequency bands could provide a neuronal basis for such discrete processes, but are rarely analyzed in this context. This article reconciles the unduly abandoned topic of discrete perception with current views and advances in neuroscience. PMID- 12757823 TI - The disunity of consciousness. AB - Attempts to decode what has become known as the (singular) neural correlate of consciousness (NCC) suppose that consciousness is a single unified entity, a belief that finds expression in the term 'unity of consciousness'. Here, I propose that the quest for the NCC will remain elusive until we acknowledge that consciousness is not a unity, and that there are instead many consciousnesses that are distributed in time and space. PMID- 12757824 TI - Constructions: a new theoretical approach to language. AB - A new theoretical approach to language has emerged in the past 10-15 years that allows linguistic observations about form-meaning pairings, known as 'constructions', to be stated directly. Constructionist approaches aim to account for the full range of facts about language, without assuming that a particular subset of the data is part of a privileged 'core'. Researchers in this field argue that unusual constructions shed light on more general issues, and can illuminate what is required for a complete account of language. PMID- 12757825 TI - Behavioural studies of strategic thinking in games. AB - Game theory is a mathematical language for describing strategic interactions, in which each player's choice affects the payoff of other players (where players can be genes, people, companies, nation-states, etc.). The impact of game theory in psychology has been limited by the lack of cognitive mechanisms underlying game theoretic predictions. 'Behavioural game theory' is a recent approach linking game theory to cognitive science by adding cognitive details about 'social utility functions', theories of limits on iterated thinking, and statistical theories of how players learn and influence others. New directions include the effects of game descriptions on choice ('framing'), strategic heuristics, and mental representation. These ideas will help root game theory more deeply in cognitive science and extend the scope of both enterprises. PMID- 12757826 TI - Animal models of periadolescent substance abuse. AB - Use of addictive substances by human juveniles and adolescents is common, including use to intoxication and progression from one drug to other drugs. Until quite recently, animal studies have not addressed the periadolescent period as a time of special vulnerability to substance abuse. For ethanol (EtOH), the most studied drug in periadolescent animals, it has become clear that effects of alcohol are similar in some ways, but different in others, compared to the effects seen in adult animals. Sparse data suggest that this conclusion may apply to other drugs as well. Recent work in rats indicates that periadolescent substance use may disrupt normal pubertal development, and may induce stronger effects on systems subserving plasticity and cognition than in adults. Animal data also indicate that some drugs may produce altered subsequent sensitivity to the same or a different drug, changes in adult cognitive capabilities, and even CNS damage. It is now clear that there can be no presumption that all studies of abusable substances carried out in adult animals will generalize readily to periadolescents. Some data suggest that continuing developmental changes in receptor expression may underlie age-related changes in drug effects. However, the biological characteristics of periadolescence which predispose toward consumption to intoxication, and the mechanisms underlying drug progression and persisting drug effects, still remain to be well defined. PMID- 12757827 TI - Brain growth spurt-prenatal ethanol exposure and the guinea pig hippocampal glutamate signaling system. AB - This study tested the hypothesis that prenatal ethanol exposure (PEE) during the brain growth spurt (BGS) in the guinea pig suppresses the glutamate-NMDA receptor nitric oxide synthase (NOS) signaling system in the developing hippocampus. Pregnant guinea pigs [term, about gestational day (GD) 68] received daily oral administration of 2 g ethanol/kg maternal body weight/day on GD 43 and/or GD 44 and then 4 g ethanol/kg maternal body weight/day from GD 45 to GD 62, isocaloric sucrose/pair-feeding or water. Offspring were studied at GD 63 (near-term fetus) and postnatal day (PD) 10 (young postnatal life). Maternal blood ethanol concentration during ethanol treatment, pregnancy outcome variables, no change in spontaneous locomotor activity, and decreased brain and cerebral cortical weight data were reported previously [Neurotoxicol. Teratol. 23 (2001) 355]. This BGS PEE regimen did not affect hippocampal stimulated glutamate release in young postnatal offspring, NMDA receptors as assessed by [3H]MK-801 binding, or NOS activity in near-term fetal offspring. Furthermore, BGS-PEE did not affect the number of hippocampal CA1 and CA3 pyramidal cells and dentate gyrus granule cells in defined locations of these three regions in the hippocampal formation. These findings are in contrast to the effects of chronic prenatal exposure to this ethanol regimen throughout gestation, including suppression of the hippocampal glutamate-NMDA receptor-NOS signaling system, decreased number of hippocampal CA1 pyramidal cells, increased spontaneous locomotor activity, and impaired performance in the Morris water maze. PMID- 12757828 TI - Behavioral response profiles following drug challenge with dopamine receptor subtype agonists and antagonists in developing rat. AB - As part of an investigation into the effects of gestational ethanol (ETOH) exposure on the developing dopamine (DA) system, pregnant Sprague-Dawley rats were exposed to one of three conditions: ETOH, pair-fed (PF) to the ETOH group, or ad libitum lab chow controls (LC). In this paper we report behavioral drug challenge effects for offspring of the two control groups (PF and LC). Male and female pups between postnatal days (PNDs) 21 and 23 in age were exposed to one of three intraperitoneal/subcutaneous doses of one of eight drugs chosen to assess the functional status of the DA D(1), D(2), and D(3) receptor subtype, or a saline control. Agonists were SKF 38393, apomorphine (APO), quinpirole (QUIN), and 7-hydroxy-N,N-di-n-propyl-2-amino-tetralin [7-OH-DPAT (DPAT)]; antagonists were spiperone (SPIP), SCH 23390, and two recently developed D(3) antagonists nafadotride (NAF) and PD 152255. Immediately following drug injection, pups were placed in observation cages, where eight behaviors (square entries, grooming, circling, rearing, sniffing, head and oral movements, and yawning) were scored at 3-min intervals for 30 min. Classic behavioral profiles were generally obtained for the high-dose mixed agonists APO, DPAT, and QUIN, which potently increased square entries, rearing, and sniffing, while reducing grooming and head movements. However, low-dose APO had no effect on behavior. The D(1) agonist, SKF 38393, had a strikingly different behavioral profile; it had no effect on square entries at any dose, while increasing grooming and sniffing at the medium dose. The D(1) antagonist, SCH 23390, profoundly decreased all behaviors except oral and head movements, especially at high doses. In contrast, the effects of the D(2) antagonist, SPIP, were limited to increasing sniffing at the medium dose. The two putative D(3) antagonists, NAF and PD 152255, presented strikingly different profiles. NAF induced a pattern of behavioral suppression that resembled the profile of high-dose SCH, while high-dose PD 152255 stimulated behavior. The failure of low-dose APO to have any effect on behavior suggests that the D(2) autoreceptor is not functional in preweanling rats. This hypothesis is further supported by the lack of behavioral suppression seen with low-dose QUIN and DPAT. Failure of NAF to produce behavioral activation at low doses and the stimulatory effects seen with PD 152255 suggests that either the D(3) autoreceptor, the postsynaptic D(3) receptor, or both are not fully functional at this age as well. PMID- 12757829 TI - Long-term nicotine exposure reduces Purkinje cell number in the adult rat cerebellar vermis. AB - Nicotine affects functions of the central nervous system. A previous study showed that developing cerebellar Purkinje cells are targets for early postnatal nicotine exposure. In this study, we assessed the effects of long-term nicotine exposure on mature cerebellar Purkinje cells. This is particularly relevant since the majority of smokers are exposed to nicotine over a long period. Female adult Sprague-Dawley rats received three doses of nicotine (0.01%, 0.03%, or 0.06%) through their sole water source. After 8 weeks of nicotine exposure, the cerebellar vermis was removed and processed for stereological cell counting. The results showed that this long-term nicotine treatment did not change the cerebellum weight or the size (volume) of the cerebellar vermis. The long-term nicotine treatment regimen did result in a significant loss of mature Purkinje cells in the cerebellum, however, such a loss of Purkinje cells was not nicotine dose-related. These findings indicated that the mature adult cerebellum is susceptible to the damaging effects of nicotine in depleting Purkinje cells in the cerebellum. PMID- 12757831 TI - Mercury (Hg2+) suppression of potassium currents of outer hair cells. AB - The heavy metal mercury (Hg(2+)) is an insidious environmental pollutant that causes toxic effects on sensory systems. It is well known that the group IIB divalent cation Hg(2+) is an inhibitor of the group I monovalent potassium (K(+)) cation pore-forming channel in several biological preparations. Here, we used the whole cell patch clamp technique on freshly isolated outer hair cells (OHCs) of the guinea pig cochlea to record outward K(+) currents and inward K(+) currents treated with mercuric chloride (HgCl(2)). HgCl(2) affected K(+) currents in a voltage- and dose-dependent manner. The effects of HgCl(2) at 1.0-100 microM are more pronounced on onset peak current than on steady-state end current. HgCl(2) depolarized also the resting membrane potential. Although the effect of HgCl(2) at 1.0 microM was partially washed out over several minutes, the effects at 10 and 100 microM were irreversible to washout. Since K(+) channels of OHCs are targets for HgCl(2) ototoxicity, this may lead to auditory transduction problems, including a loss in hearing sensitivity. A better understanding of fundamental mechanisms underlying K(+) channelopathies in OHCs due to HgCl(2) poisoning may lead to better preventive or therapeutic agents. PMID- 12757830 TI - Neurobehavioral changes in mice chronically exposed to methylmercury during fetal and early postnatal development. AB - Pregnant C57BL/6 mice were chronically treated with 0, 4, 6, or 8 ppm of methylmercury chloride (MeHg) in drinking water during fetal and early postnatal development. Four behavioral functions were analyzed in female and male offspring between the age of 6 and 12 weeks: motor coordination learning on the rotarod; training to spatial alternation in the standard T maze followed by a working memory test with delays; spontaneous locomotion and rearings in the open field; reference and working memory assessment in the modified T maze [Behav. Neurosci. 102 (1988) 635]. Chronic perinatal treatment with MeHg resulted in moderate brain levels of mercury near birth which rapidly decreased during nursing. MeHg exerted an effect on the performance of females, but not of males, on two of the four measurements. All treated females exhibited less locomotion than control mice when the open field was new, but not in the following four sessions when the environment was becoming increasingly familiar. Working memory was impaired in females treated with 6 and 8 ppm of MeHg in the modified T maze, but not on the test with delays in the standard T maze. Taken together, these results show that chronic exposure to MeHg during fetal and postnatal development had sex-dependent effects on horizontal exploration and on working memory in the modified T maze, and no effects on motor coordination learning and reference memory. PMID- 12757832 TI - Effects of alcohol on cocaine lethality in rats: acute and chronic assessments. AB - Cocaine and alcohol is a popular, yet toxic, drug combination that results in effects greater than that of either drug alone. The following experiment presents additional evidence that supports this position. Specifically, the lethal effects of acute (1 day) and chronic (4 days) alcohol (0.5 g/kg), cocaine (20, 30, or 40 mg/kg), or the respective alcohol/cocaine combinations were assessed in rats. For acute drug administration, lethality was only evident in those animals administered the combination of 0.5 g/kg alcohol and 40 mg/kg cocaine, supporting the position that the effects of combining alcohol and cocaine are greater than either drug alone. Chronic drug administration resulted in a weak sensitization to cocaine-induced lethality that was increased when alcohol was combined with cocaine. Together, this evidence suggests that combining alcohol and cocaine (acutely) can have lethal consequences in rats that are greater than either drug alone, effects that can be exacerbated with chronic use. PMID- 12757833 TI - Effects of recombinant human growth hormone on the pharmacokinetics of intravenous chlorzoxazone in rats with acute renal failure induced by uranyl nitrate. AB - It has been reported from our laboratories that expression of CYP2E1 significantly increased and decreased in rats with acute renal failure induced by uranyl nitrate (U-ARF) treated with recombinant human growth hormone (rGH) for one day (U-ARF1) compared with those in control rats and rats with U-ARF, respectively. Chlorzoxazone (CZX) primarily undergoes hydroxylation to form 6 hydroxychlorzoxazone (OH-CZX) catalyzed mainly by CYP2E1 in rats. Hence, the effects of rGH on the pharmacokinetics of intravenous CZX (20 mg/kg) were investigated in rats with U-ARF. Based on CYP2E1 expression, it could be expected that in rats with U-ARF1, the formation of OH-CZX significantly increased and decreased compared with those in control rats and rats with U-ARF, respectively. This was proven in the following results. First, the total area under the plasma concentration-time curve from time zero to 8 hr (AUC(0-->8 hr)) of OH-CZX in rats with U-ARF1 (36,100 microg min/ml) was significantly greater and smaller than those in control rats (1040 microg min/ml) and rats with U-ARF (50,300 microg min/ml), respectively. Second, the AUC(0-->8 hr, OH-CZX)/AUC(CZX) ratio in rats with U-ARF1 (28.9) was significantly greater and smaller than those in control rats (0.468) and rats with U-ARF (72.6), respectively. PMID- 12757834 TI - An androgen receptor in the brain of the green frog Rana esculenta. AB - An androgen receptor (AR) has been detected in both the hypothalamic and extra hypothalamic area of the brain of the male of the green frog Rana esculenta. 3H-T binding activity was detected separately in the hypothalamic and extra hypothalamic areas. Experiments of binding assay showed 3H-T binding activity in the nuclear extract but not in the cytosol of both the hypothalamic and the extra hypothalamic areas. The androgen binding moiety was not strictly specific for androgens, binding also 17 beta-estradiol, although to a lesser extent. 3H testosterone binding activity fluctuated in both the hypothalamic and the extra hypothalamic areas throughout the reproductive cycle, and paralleled androgen plasma levels. PMID- 12757835 TI - Preliminary evaluation of several disinfection/sterilization techniques for use with microdialysis probes. AB - This study evaluated the suitability of some disinfection and sterilization methods for use with microdialysis probes. Disinfection or sterilization should minimize the tissue inflammatory reaction and improve the long-term health of rats on study and ensure the quality of data obtained by microdialysis sampling. Furthermore, the treatment should not negatively impact probe integrity or sampling performance. The techniques chosen for evaluation included two disinfection methods (70% ethanol and a commercial contact lens solution) and two sterilization methods (hydrogen peroxide plasma, and e-beam radiation). Linear microdialysis probes treated by these processes were compared to untreated probes removed from the manufacturer's packaging as if sterile (the control group). The probes were aseptically implanted in the livers of rats and monitored for 72 hours. The parameters chosen to evaluate probe performance were relative sample mass recovery and the relative in vivo extraction efficiency of the probe for caffeine. Post mortem bacterial counts and histopathology examination of liver tissue were also conducted. The probes remained intact and functional for the entire study period. The methods tested did not acutely alter the probes although hydrogen peroxide plasma and contact lens solution groups showed reduced extraction efficiencies. Minimal tissue damage was observed surrounding the probes and acute inflammatory reaction was mild to moderate. Low numbers of bacterial colonies from the implantation sites indicates that the health of animals in this study was not impaired. This was also true for the control group (untreated probe). PMID- 12757836 TI - Pro-oxidant effect of dehydroepiandrosterone in rats is mediated by PPAR activation. AB - DHEA-treatment exerts a dual effect, prooxidant or antioxidant, depending on the dosage and, therefore, on the tissue concentration reached. In agreement with previous studies showing a prooxidant effect of DHEA, here we show that pharmacological doses of DHEA produce increased H(2)O(2) levels and a marked reduction of GSH content in rat liver. DHEA, also increases both catalase (by 30%) and cytochrome-C-reductase (by 30%) activities in the liver cytosol. The effectiveness of the state of increased oxidative stress is also documented by changes in fatty acid pattern of the microsomal membranes. Moreover, DHEA, at high doses, enhances beta-oxidation, as demonstrated by an increase of acyl-CoA oxidase activity and of cytochrome P450 4A content, confirming that it acts as a PPARs inducer. Both PPARs induction and proxidant effects completely disappear when DHEA is administered at lower doses. Seven days treatment (4 or 10 mg) is unable to affect either levels of proxidant species and of antioxidant molecules, or cytochrome P450 4A content and beta-oxidation. Prolonged DHEA treatment (4 mg/day) for three weeks not only is unable to affect PPARs activation and beta oxidation, but it also exerts a protective effect against ADP/Fe(2+) induced lipid peroxidation. This latter result confirms the antioxidant effects of DHEA at low doses, as already previously documented. PMID- 12757837 TI - Thrombin-induced Ca2+ mobilization in human gingival fibroblasts is mediated by protease-activated receptor-1 (PAR-1). AB - By reverse transcription (RT)-PCR analyses, human gingival fibroblasts (HGFs) were demonstrated to express mRNAs for protease-activated receptor-1 (PAR-1) and PAR-3, although the expression of PAR-3 was much weaker than that of PAR-1. The mRNAs for PAR-2 and PAR-4 were not found by RT-PCR. Furthermore, PAR activation was studied by monitoring cytosolic Ca(2+) concentration ([Ca(2+)]i) in cultured HGFs loaded with fura-2. At concentrations > 0.1 nM, alpha-thrombin caused a transient increase in [Ca(2+)]i in a concentration-dependent manner, and the maximum response was obtained with 10 nM alpha-thrombin. After the [Ca(2+)]i response, the HGFs were completely desensitized to the second stimulation with alpha-thrombin. The PAR-1 agonist peptide SFLLRN produced approximately the same transient [Ca(2+)]i response as alpha-thrombin. After stimulation with SFLLRN, the HGFs did not respond to alpha-thrombin, indicating that treatment with SFLLRN results in complete desensitization to alpha-thrombin. The PAR-2 and PAR-4 agonist peptides had no effect on [Ca(2+)]i in HGFs. These results suggest that alpha-thrombin-induced Ca(2+) mobilization in HGFs is solely mediated by PAR-1. PMID- 12757838 TI - Effects of nicotine on target biting and resident-intruder attack. AB - The effects of acute administration of nicotine on target biting (defensive) and resident-intruder (offensive) attack of male mice were assessed. In the target biting procedure confined mice received tail shock on a fixed time, 2-min schedule. Under baseline conditions, biting attack directed toward an inanimate target occurred at three distinct rates. A high target biting rate (13.5 +/- 3.8 bites/15 sec) followed shock delivery, an intermediate biting rate (9.6 +/- 4.1 bites/15 sec) occurred during the inter-shock interval, and a low biting rate (1.0 +/- 0.5 bites/15 sec) occurred during a tone stimulus which signalled the impending shock. Nicotine (administered IP, 15 min presession) reduced post-shock and inter-shock interval target biting in a dose-dependent manner (ED50 values estimated at 0.13 and 0.14 mg/kg, respectively) but exerted more variable effects on target biting during the tone. In the resident-intruder paradigm the same mice were exposed to an intruder introduced into its home cage for a 10-min test session. Under baseline conditions, residents directed 20 +/- 3.2 biting attacks toward the intruder during the session with an average latency of 89 +/- 40 sec to the first attack. Nicotine caused a dose-dependent decrease in this attack behavior (ED50 values estimated to be 0.48 and 0.49 mg/kg, respectively). These observations are interpreted to indicate that nicotine has an increased potency at reducing "defensive" aggression. PMID- 12757839 TI - Effect of age on body distribution of Tityustoxin from Tityus serrulatus scorpion venom in rats. AB - Previous research from our Laboratory has shown a greater susceptibility of young animals, when compared to adults, to envenomation by tityustoxin (TsTX), one of the main toxins from Tityus serrulatus scorpion venom. Our hypothesis is that a differential body distribution of TsTX among adult and young animals could account for the worse prognosis of scorpion envenomation in infants. Thus, TsTX labeled with technetium-99m was injected (6 microg, subcutaneous) in adult (150 160 day-old) and young (21-22 day-old) male rats. Groups of animals were sacrificed at different times after TsTX injection (0.08, 1.0, 3.0, 6.0, 12.0 and 24.0 hours) under Urethane anesthesia (140 mg/100 g, i.p.). The brain, heart, lungs, liver, kidneys, spleen and thyroid were excised and blood collected. Young rats presented a shorter latency toxin concentration peak in all studied organs except for the liver and the kidney, when compared to adults. The ratio between the area under the curve of the toxin concentration in each organ and that in blood (Kp) indicates higher accumulation in the organs of young animals mainly for brain, liver and heart. These observations suggest a faster toxin distribution in the organs of young rats. The higher uptake of TsTX in the brain is suggestive of a greater permeability for the toxin along the blood-brain barrier of young rats. In conclusion, the higher uptake in heart, together with data from the brain, may help to elucidate the clinical manifestations frequently observed in children under scorpion envenomation. PMID- 12757840 TI - Effect of age and photoperiodic conditions on metabolism and oxidative stress related markers at different circadian stages in rat liver and kidney. AB - It has been shown that some cytochrome P450-dependent enzyme activities could present daily fluctuations, particularly CYP3A isoenzymes which are enhanced during the dark period. The aim of this study was to investigate whether age and photoperiodic conditions at different circadian stages could influence these fluctuations. Young mature (10 weeks) and old (22 months) Wistar rats were initially exposed to light-dark cycles 12:12 during 4 weeks, and secondly 18:6 for either one week or six weeks. Erythromycin N-demethylase (CYP3A-dependent), 7 ethoxycoumarin O-deethylase (CYP1A-dependent) and aniline 4-hydroxylase (CYP2E dependent) activities were determined in liver and kidney microsomes at different hours after darkness onset (HADO). In addition, liver and kidney GSH, GSHPx, ATP, TBARS were determined. During the LD 12:12 cycle, while no significant modification was observed in CYP1A- and 2E-dependent enzyme activities as functions of HADO, erythromycin N-demethylase activity (CYP3A-dependent) showed a significant increase during the second third of the dark period in both young and old rats. After switching to a LD 18:6 cycle, this variation was still observed during second third of the dark period, to a lesser but still significant degree, with no difference between one week and six weeks exposure to the new photoperiod. It can be noted that the old rats showed a significantly lower level of erythromycin N-demethylase activity than the young rats, in parallel to a decrease in GSH, GSHPx and ATP, and an increase in TBARS. These results confirm the lower resistance of old animals to oxidative stress. The observed variations in metabolism parameters underline the need for study designs in pharmaco toxicology taking into account the possible risks induced by circadian changes, especially in aged subjects. PMID- 12757841 TI - Eugenol suppresses cyclooxygenase-2 expression in lipopolysaccharide-stimulated mouse macrophage RAW264.7 cells. AB - Inducible cyclooxygenase (COX-2) has been implicated in the processes of inflammation and carcinogenesis. Thus, the potential COX-2 inhibitors have been considered as anti-inflammatory or cancer chemopreventive agents. In this study, the methanolic extract of the cortex of Eugenia caryophyllata Thunberg (Myrtaceae) was found to potently inhibit the prostaglandin E(2) production in lipopolysaccharide (LPS)-activated mouse macrophage RAW264.7 cells (98.3% inhibition at the test concentration of 10 microg/ml). Further, hexane-soluble layer was the most active partition compared to ethyl acetate, n-butanol, and water-soluble parts. By bioassay-guided fractionation of hexane-soluble partition, eugenol was isolated and exhibited a significant inhibition of PGE(2) production (IC(50) = 0.37 microM). In addition, eugenol suppressed the cyclooxygenase-2 (COX-2) gene expression in LPS-stimulated mouse macrophage cells. On the line of COX-2 playing an important role in colon carcinogenesis further study was designed to investigate the effect of eugenol on the growth and COX-2 expression in HT-29 human colon cancer cells. Eugenol inhibited the proliferation of HT-29 cells and the mRNA expression of COX-2, but not COX-1. This result suggests that eugenol might be a plausible lead candidate for further developing the COX-2 inhibitor as an anti-inflammatory or cancer chemopreventive agent. PMID- 12757842 TI - Relationship between 24-hour urinary-free cortisol excretion and salivary cortisol levels sampled from awakening to bedtime in healthy subjects. AB - The utility of repeated salivary cortisol sampling as a substitute for 24-hour urinary-free cortisol (UFC) assessment was examined. Forty-four participants completed both 24-hour collections and 6 salivary collections at wake-up, 08:00, 12:00, 16:00, 20:00 and bedtime, during the same 24-hour period. The results demonstrated that mean, maximum, and amplitude (maximum minus minimum) for salivary cortisol all correlated positively with urinary cortisol, but the associations of these variables with urinary-free cortisol excretion were relatively small. Furthermore, a single salivary sample taken at wake-up was as good an indicator of overall cortisol production as the measures derived from multiple salivary samples. An examination of subject compliance indicated that many subjects failed to collect the timed salivary collections as instructed. The authors conclude that diurnal salivary cortisol sampling versus 24-hour urinary cortisol collections are likely to provide different information about ambient hypothalamic-pituitary-adrenal productivity, and therefore these measures should not be used interchangeably. In addition, subject compliance is a serious consideration in designing studies that employ home salivary collections. PMID- 12757843 TI - Involvement of cytochrome P450 and the flavin-containing monooxygenase(s) in the sulphoxidation of simple sulphides in human liver microsomes. AB - This study was conducted to examine the involvement of cytochrome P450 (CYP450) and the flavin-containing monooxygenase (FMO) in the sulphoxidation of ethyl methyl sulphide (EMS), 4-chlorophenyl methyl sulphide (CPMS) and diphenyl sulphide (DPS) in human liver microsomes from a phenotypic CYP2D6 extensive metabolizer. Human liver microsomes catalyzed the sulphoxidation of EMS, CPMS and DPS to their corresponding sulphoxides. Lineweaver-Burk plots for the sulphoxidation of EMS in human liver microsomes indicated that the apparent K(m) and V(max) were 1.53 +/- 0.07 mM and 1.11 +/- 0.25 nmoles/mg protein/min, respectively. The apparent K(m) and V(max) for the sulphoxidation of CPMS were 0.17 +/- 0.05 mM and 1.41 +/- 0.16 nmoles/mg protein/min, respectively. The apparent K(m) and V(max) for the sulphoxidation of DPS were 0.10 +/- 0.01 mM and 1.08 +/- 0.05 nmoles/mg protein/min, respectively. Methimazole noncompetitively inhibited the sulphoxidation of EMS, CPMS and DPS by human liver microsomes with K(i) values of 8.6 +/- 0.6, 5.7 +/- 0.4 and 6.6 +/- 0.5 mM, respectively. SKF525A noncompetitively inhibited the sulphoxidation of CPMS and DPS by human liver microsomes with K(i) values of 6.6 +/- 0.4 and 0.40 +/- 0.1 mM, respectively. The results suggest that FMO is involved in the sulphoxidation of EMS, CPMS and DPS while CYP450 is involved in the sulphoxidation of CPMS and DPS in human liver microsomes. PMID- 12757844 TI - In vitro characterization of [3H]MethoxyPyEP, an mGluR5 selective radioligand. AB - We have characterized the in vitro properties of 3-[3H]methoxy-5-(pyridin-2 ylethynyl)pyridine ([3H]MethoxyPyEP), an analogue of the mGluR(5) receptor subtype antagonist MPEP [2-methyl-6-(phenylethynyl)-pyridine], in rat tissue preparations using tissue homogenates and autoradiography. Binding of [3H]MethoxyPyEP to rat cortex, hippocampus, thalamus and cerebellum membrane preparations revealed saturable, high affinity binding (3.4 +/- 0.4 nM, n = 4 in rat cortex) to a single population of receptors in all regions studied except for cerebellum. Binding was found to be relatively insensitive to pH and insensitive to DTT. High concentrations of NEM both reduce receptor concentration and binding affinity for the radioligand. In time-course studies at room temperature k(on) and k(off) were determined as 2.9 x 10(7) M(-1) min(-1) and 0.11 min(-1) respectively. The rank order of affinities, as assessed by equilibrium competition studies, of a variety of ligands suggested binding of the radioligand selectively to mGluR5 (MPEP > trans-azetidine-2,4-dicarboxylic acid congruent with (S)-4-carboxyphenylglycine congruent with (+)MK801 congruent with CP-101,606 congruent with clozapine congruent with atropine congruent with ketanserin congruent with yohimbine congruent with benoxathian). Autoradiographic studies with [3H]MethoxyPyEP showed that binding was regioselective, with high density of binding in caudate and hippocampus, intermediate binding in thalamus and very low density in the cerebellum. These data show that [3H]MethoxyPyEP is a high affinity radioligand useful for the in vitro study of mGluR5 receptor distribution and pharmacologic properties in brain. PMID- 12757845 TI - Editorial commentary on "Superoxide reacts with hydroethidine but forms a fluorescent product that is distinctly different from ethidium: potential implications in intracellular fluorescence detection of superoxide" by H. Zhao et al. PMID- 12757846 TI - Superoxide reacts with hydroethidine but forms a fluorescent product that is distinctly different from ethidium: potential implications in intracellular fluorescence detection of superoxide. AB - Hydroethidine (HE) or dihydroethidium (DHE), a redox-sensitive probe, has been widely used to detect intracellular superoxide anion. It is a common assumption that the reaction between superoxide and HE results in the formation of a two electron oxidized product, ethidium (E+), which binds to DNA and leads to the enhancement of fluorescence (excitation, 500-530 nm; emission, 590-620 nm). However, the mechanism of oxidation of HE by the superoxide anion still remains unclear. In the present study, we show that superoxide generated in several enzymatic or chemical systems (e.g., xanthine/xanthine oxidase, endothelial nitric oxide synthase, or potassium superoxide) oxidizes HE to a fluorescent product (excitation, 480 nm; emission, 567 nm) that is totally different from E+. HPLC measurements revealed that the HE/superoxide reaction product elutes differently from E+. This new product exhibited an increase in fluorescence in the presence of DNA. Mass spectral data indicated that the molecular weight of the HE/superoxide reaction product is 330, while ethidium has a molecular weight of 314. We conclude that the reaction between superoxide and HE forms a fluorescent marker product that is different from ethidium. Potential implications of this finding in intracellular detection and imaging of superoxide are discussed. PMID- 12757847 TI - Oxidative stress and calcium signaling in the adverse effects of environmental particles (PM10). AB - This review focuses on the potential role that oxidative stress plays in the adverse effects of PM(10). The central hypothesis is that the ability of PM(10) to cause oxidative stress underlies the association between increased exposure to PM(10) and both exacerbations of lung disease and lung cancer. Pulmonary inflammation may also underlie the cardiovascular effects seen following increased PM(10), although the mechanisms of the cardiovascular effects of PM(10) are not well understood. PM(10) is a complex mix of various particle types and several of the components of PM(10) are likely to be involved in the induction of oxidative stress. The most likely of these are transition metals, ultrafine particle surfaces, and organic compounds. In support of this hypothesis, oxidative stress arising from PM(10) has been shown to activate a number of redox responsive signaling pathways in lung target cells. These pathways are involved in expression of genes that play a role in responses relevant to inflammation and pathological change, including MAPKs, NF-kappaB, AP-1, and histone acetylation. Oxidative stress from particles is also likely to play an important role in the carcinogenic effects associated with PM(10) and hydroxyl radicals from PM(10) cause DNA damage in vitro. PMID- 12757848 TI - Mutant Cu,Zn superoxide dismutases and familial amyotrophic lateral sclerosis: evaluation of oxidative hypotheses. AB - FALS-associated missense mutations of SOD1 exhibit a toxic gain of function that leads to the death of motor neurons. The explanations for this toxicity fall into two broad categories. One involves a gain of some oxidative activity, while the second involves a gain of protein: protein interactions. Among the postulated oxidative activities are the following: (i) peroxidase action; (ii) superoxide reductase action; and, (iii) the enhancement of production of O2- by partial reversal of the normal SOD activity, which then leads to increased formation of ONOO(-). We will herein concentrate on evaluating the relative merits of these oxidative hypotheses and consider whether the experiments with transgenic animals that purport to disprove these oxidative explanations really do so. PMID- 12757849 TI - HO-1 induction attenuates renal damage and oxidative stress induced by K2Cr2O7. AB - Heme oxygenase (HO) is the rate-limiting enzyme in the degradation of heme; its inducible isozyme HO-1 protects against some types of acute tissue injury. The expression and functional role of HO-1 in rats with renal injury induced by potassium dichromate (K(2)Cr(2)O(7)) was investigated in this work. Rats were studied 24 h after a single injection of K(2)Cr(2)O(7). To address the possible protective effect of HO-1 in this experimental model, this enzyme was induced by an injection of stannous chloride (SnCl(2)) 12 h before K(2)Cr(2)O(7) administration. The functional role of HO-1 in K(2)Cr(2)O(7) + SnCl(2)-treated animals was tested by inhibiting HO activity with an injection of zinc (II) protoporphyrin IX (ZnPP) 18 h before K(2)Cr(2)O(7). In K(2)Cr(2)O(7)-treated rats: (i) renal HO-1 content, measured by Western blot, increased 2.6-fold; and, (ii) renal nitrotyrosine and protein carbonyl content, markers of oxidative stress, increased 3.5- and 1.36-fold, respectively. Renal damage and oxidative stress were ameliorated and HO-1 content was increased in the K(2)Cr(2)O(7) + SnCl(2) group. The attenuation of renal injury and oxidative stress was lost by the inhibition of HO activity in K(2)Cr(2)O(7) + SnCl(2) + ZnPP-treated animals. Our data suggest that HO-1 overexpression induced by SnCl(2) is responsible for the attenuation of renal damage and oxidative stress induced by K(2)Cr(2)O(7). PMID- 12757850 TI - The mode of decomposition of Angeli's salt (Na2N2O3) and the effects thereon of oxygen, nitrite, superoxide dismutase, and glutathione. AB - The classical view of the aerobic decomposition of Angeli's salt is that it releases NO(2)(-) + NO(-)/HNO the latter then reacting with O(2) to yield ONOO( ). An alternative that has recently been proposed envisions electron transfer to O(2) followed by decomposition to NO(2)(-) + NO. The classical view is now strongly supported by the observation that the rates of decomposition of Angeli's salt under 20% O(2) or 100% O(2) were equal. Moreover, NO(2)(-), which inhibits this decomposition by favoring the back reaction, was more effective in the absence of agents that scavenge NO(-)/HNO. It is thus clear that Angeli's salt is a useful source of NO(-)/HNO for use in defined aqueous systems. The measurements made in the course of this work allowed approximation of the rate constants for the reactions of NO(-)/HNO with NO(2)(-), O(2), glutathione, or Cu, Zn superoxide dismutase. The likelihood of the formation of NO(-)/HNO in vivo is also discussed. PMID- 12757852 TI - Zinc-induced apoptosis in substantia nigra of rat brain: neuroprotection by vitamin D3. AB - Accumulation of transition metals has been suggested to be responsible for the deteriorated nigrostriatal dopaminergic system in Parkinson's patients. In the present study, the mechanism underlying the zinc-induced neurotoxicity was investigated in the nigrostriatal dopaminergic system in vivo. Our 6-methoxy-8 paratoluene sulfonamide quinoline fluorescence study showed zinc translocation in the infused nigral cells after intranigral infusion of zinc. Furthermore, lipid peroxidation in the zinc-infused substantia nigra was consistently elevated 4 h to 7 d after the infusion. At the same time, an abrupt increase in cytosolic cytochrome c content in the infused substantia nigra was observed 4 h after zinc infusion and gradually decreased to basal levels 7 d after infusion. Both TUNEL positive neurons and DNA fragmentation, indicatives of apoptosis, were detected in the zinc-infused substantia nigra. Furthermore, striatal dopamine content was reduced 7 d after the infusion. In attempt to prevent zinc-induced neurotoxicity, vitamin D3 was systemically administered. Zinc-induced increases in lipid peroxidation and cytosolic cytochrome c in the infused substantia nigra were prevented by this treatment. Moreover, zinc-induced reduction in striatal dopamine content was attenuated after vitamin D3 treatment. Our in vivo data suggest that zinc-induced oxidative stress may result in apoptosis followed by reduced dopaminergic function in the nigrostriatal dopaminergic system. Furthermore, vitamin D3 prevented zinc-induced oxidative injuries in the rat brain. PMID- 12757851 TI - A stress-responsive glutathione S-transferase confers resistance to oxidative stress in Caenorhabditis elegans. AB - Previous studies demonstrated that the Caenorhabditis elegans GST-p24 is upregulated at the steady state mRNA level in response to oxidative stress. A transcriptional upregulation was confirmed in the current study by analyzing Ce GST-p24 promoter-reporter constructs in transgenic C. elegans strains CL2166 and CL3166. The transgenic strain BL1, which overexpresses the Ce-GST-p24 enzyme (as a GFP fusion protein controlled by its own promoter), was generated to investigate the function of this enzyme in vivo. Stress experiments with BL1 demonstrated an increased resistance to intracellularly induced oxidative stress, as compared to wild type. The consequences of a decrease in the Ce-GST-p24 enzyme concentration were examined by RNAi-treatment of BL1 C. elegans to silence both the endogene and the transgene Ce-GST-p24 and by the analysis of the K08F4.7 homozygous deletion mutant. In both cases, the reduced Ce-GST-p24 enzyme level resulted in a significant decrease in the stress resistance of the nematodes. These results clearly demonstrate a direct correlation between the concentration of Ce-GST-p24 and the resistance to oxidative stress. We have demonstrated for the first time that manipulation of the expression of a single GST can modulate the organismal response to oxidative stress. The enzymatic activity of this detoxification enzyme was examined with various substrates, giving emphasis to lipid peroxidation products. The Ce-GST-p24 was also localized in BL1 C. elegans by confocal laser-scanning microscopy, revealing a wide-spread distribution profile. PMID- 12757853 TI - Saccharated colloidal iron enhances lipopolysaccharide-induced nitric oxide production in vivo. AB - We investigated the effects of iron on the production of nitric oxide (NO), inducible NO synthase (iNOS), and plasma cytokines induced by lipopolysaccharide (LPS) in vivo. Male Wistar rats were preloaded with a single intravenous injection of saccharated colloidal iron (Fesin, 70 mg iron/kg body weight) or normal saline as a control, and then given an intraperitoneal injection of LPS (5.0 mg/kg body weight). Rats, preloaded with iron, had evidence of both iron deposition and strong iNOS induction in liver Kupffer cells upon injection of LPS; phagocytic cells in the spleen and lung had similar findings. LPS-induced NO production in iron-preloaded rats was significantly higher than control rats as accessed by NO-hemoglobin levels measured by ESR (electron spin resonance) and NOx (nitrate plus nitrite) levels. Western blot analysis showed that iron preloading significantly enhanced LPS-induced iNOS induction in the liver, but not in the spleen or lung. LPS-induced plasma levels of IL-6, IL-1beta, and TNF alpha were also significantly higher in iron-preloaded rats as shown by ELISA, but IFN-gamma levels were unchanged. We conclude that colloidal-iron phagocytosed by liver Kupffer cells enhanced LPS-induced NO production in vivo, iNOS induction in the liver, and release of IL-6, IL-1beta, and TNF-alpha. PMID- 12757854 TI - The effect of valproic acid on hepatic and plasma levels of 15-F2t-isoprostane in rats. AB - The mechanism by which valproic acid (VPA) induces liver injury remains unknown, but it is hypothesized to involve the generation of toxic metabolites and/or reactive oxygen species. This study's objectives were to determine the effect of VPA on plasma and hepatic levels of the F(2)-isoprostane, 15-F(2t)-IsoP, a marker for oxidative stress, and to investigate the influence of cytochrome P450- (P450 ) mediated VPA biotransformation on 15-F(2t)-IsoP levels in rats. In rats treated with VPA (500 mg/kg), plasma 15-F(2t)-IsoP was increased 2.5-fold at t(max) = 0.5 h. Phenobarbital pretreatment (80 mg/kg/d for 4 d) in VPA-treated rats increased plasma and liver levels of free 15-F(2t)-IsoP by 5-fold and 3-fold, respectively, when compared to control groups. This was accompanied by an elevation in plasma and liver levels of P450-mediated VPA metabolites. Pretreatment with SKF-525A (80 mg/kg) or 1-aminobenzotriazole (100 mg/kg), which inhibited P450-mediated VPA metabolism, did not attenuate the increased levels of plasma 15-F(2t)-IsoP in VPA treated groups. Plasma and hepatic levels of 15-F(2t)-IsoP were further elevated after 14 d of VPA treatment compared to single-dose treatment. Our data indicate that VPA increases plasma and hepatic levels of 15-F(2t)-IsoP and this effect can be enhanced by phenobarbital by a mechanism not involving P450-catalyzed VPA biotransformation. PMID- 12757855 TI - Expression of human MutT homologue (hMTH1) protein in primary non-small-cell lung carcinomas and histologically normal surrounding tissue. AB - In situ, oxidation of deoxyguanosine yields 8-hydroxy-2'-deoxyguanosine (8-oxo dG), which is mutation prone and results in a G:C --> T:A transversion following DNA replication. Another pathway to the formation of DNA containing 8-oxo-dG is by the misincorporation of 8-oxo-dGTP via DNA polymerase. Human MutT homologue (hMTH1), an 8-oxo-dGTPase, prevents misincorporation of this oxidized nucleotide by hydrolyzing 8-oxo-dGTP to 8-oxo-dGMP. Previous studies have shown that hMTH1 mRNA is overexpressed in human renal cell carcinomas and breast tumors. Elevated levels of hMTH1 protein have also been detected in brain tumors. In the current study, we determined whether hMTH1 protein is overexpressed in primary non-small cell lung carcinomas as compared to adjacent histologically normal lung tissue. Twenty matched human lung tumor/normal pairs were examined by Western analysis for expression of hMTH1 protein. Overexpression in the tumors was detected in 4/8 (50%) adenocarcinomas, 4/4 (100%) adenocarcinomas with bronchioalveolar (BAC) features, 2/2 (100%) BACs, and 3/6 (50%) squamous cell carcinomas. The data from Western analysis were validated by immunohistochemical staining for hMTH1 protein. The results of this study indicate that hMTH1 protein may be a potential marker for the detection of persistent oxidative stress in lung cancer. PMID- 12757856 TI - Cloning of novel human SEC14p-like proteins: ligand binding and functional properties. AB - We describe the cloning and expression of two novel genes highly similar to the tocopherol-associated protein (hTAP/SEC14L2/SPF). Immunoprecipitation of the three recombinant hTAPs and extraction of their associated lipid-soluble molecules indicates that they bind not just tocopherols, but also phosphatidylinositol, phosphatidylcholine, and phosphatidylglycerol. Ligand competition analysis by isoelectric point mobility shift assay indicates that phosphatidylcholine, tocopherols, and tocopheryl-succinate compete with phosphatidylinositol binding to hTAPs. To investigate a possible function of hTAPs on enzymes involved in phospholipids metabolism, the activity of recombinant phosphatidylinositol 3-kinase (PI3Kgamma/p110gamma) was tested. Recombinant hTAPs reduce in vitro the activity of the recombinant catalytic subunit of PI3Kgamma and stimulate it in the presence of alpha-tocopherol up to 5 fold. Immunoprecipitation of hTAP1 from cells results in co-precipitation of PI3 kinase activity, indicating a physical contact between the two proteins at a cellular level. In summary, hTAPs may modulate, in a tocopherol-sensitive manner, phosphatidylinositol-3-kinase, a central enzyme in signal transduction, cell proliferation, and apoptosis. It is possible that other phosphatidylinositol- and phosphatidylcholine-dependent signaling pathways are modulated by hTAPs and tocopherols, possibly by transporting and presenting these ligands to the corresponding enzymes. PMID- 12757857 TI - Spin traps: in vitro toxicity and stability of radical adducts. AB - We have evaluated the effects of DMPO, CMPO, EMPO, BMPO, and DEPMPO on functioning CHO cells and the stability of the radical adducts in the presence of cells. The potential toxic effects of the spin traps were measured by two estimates of cell viability (trypan blue exclusion and colony formation) and one of cell function (rate of oxygen consumption). We also studied the effects of the spin traps on colony formation in a second cell line, 9L tumor cells. Toxicity varied with the type of cell line and the parameter that was measured. In aqueous solutions the order of stability for all spin adducts was SO(3) > OH > CH(3), while in cell suspensions it was SO(3) > OH approximately CH(3). The radical adducts of the new spin traps have significantly increased stability as compared to DMPO. These results indicate that the new spin traps potentially offer increased stability of spin adducts in functioning cells. It also is clear that it is necessary to carry out appropriate studies of the stability and toxicity in the system that is to be studied for any particular use of these spin traps. It then should be feasible to select the spin trap(s) best suited for the proposed study. PMID- 12757858 TI - L-2-oxothiazolidine-4-carboxylate supplementation in murine gamma-GT deficiency. AB - gamma-glutamyl transpeptidase (gamma-GT) deficiency in GGT(enu1) mice is associated with glutathionemia, glutathionuria, growth retardation, infertility, lethargy, cataracts, and shortened life span. Total liver glutathione (GSH) content is significantly reduced in gamma-GT-deficient mice due to chronic excessive GSH loss. Oral supplementation of GGT(enu1) mice with L-2 oxothiazolidine-4-carboxylate (OTZ), a cysteine prodrug, led to partial restoration of liver GSH content. The growth, physical appearance, and behavior of gamma-GT-deficient mice were substantially improved following OTZ supplementation. Tissue GSH deficiency is the proximate cause of the phenotypic abnormalities associated with murine gamma-GT deficiency. PMID- 12757859 TI - Modeling the influence of superoxide dismutase on superoxide and nitric oxide interactions, including reversible inhibition of oxygen consumption. AB - A mathematical mass transport model was constructed in cylindrical geometry to follow coupled biochemical reactions and diffusion of oxygen, nitric oxide, superoxide, peroxynitrite, hydrogen peroxide, nitrite, and nitrate around a blood vessel. Computer simulations were performed for a 50 microm internal diameter arteriole to characterize mass transport in five concentric regions (blood, plasma layer, endothelium, vascular wall, perivascular tissue). Steady state gradients in nitric oxide, oxygen partial pressure, superoxide, and peroxynitrite, and associated production of hydrogen peroxide, nitrite, and nitrate were predicted for varying superoxide production rates, superoxide dismutase concentrations, and other physiological conditions. The model quantifies how competition between superoxide scavenging by nitric oxide and superoxide dismutase catalyzed removal varies spatially. Reversible inhibition of oxygen consumption by nitric oxide, which causes increased tissue oxygenation at deeper locations, was also included in the model. The mass transport model provides insight into complex interactions between reactive oxygen and nitrogen species in blood and tissue, and provides an objective way to evaluate the relative influence of different biochemical pathways on these interactions. PMID- 12757860 TI - Spotlight on stem cells--makes old hearts fresh. PMID- 12757861 TI - Regulatory modules in the developing heart. AB - Fragments of regulatory DNA of cardiac genes drive reporter gene expression in sometimes unexpected subdomains of the heart. These patterns have revealed that the regulatory DNA of genes consists of distinct subfragments (regulatory modules) that are active in different regions of the developing heart. In this review we give an overview of the activity of regulatory modules in vivo. Furthermore, we investigated the relationship between the activity domains of the regulatory modules, the building blocks of the heart and the developmental patterning of the myocardium. Most of the regulatory modules show a domain of activity broader than the morphological boundary of a cardiac compartment and seem to respond to a patterning program along the antero-posterior axis. PMID- 12757862 TI - Molecular pathways in myocardial development: a stem cell perspective. AB - The heart has long been considered to adapt to increased work or pathology through the cellular growth process of hypertrophy. However, recent evidence for the existence of endogenous stem cells and regenerative capacity in the adult heart has given new impetus to the quest for cell therapies for heart failure, which remains the number one killer in Western cultures. The molecular cues driving cardiac development are now being explored in detail and will come into sharp focus as regimes for stem cell differentiation and efforts to augment endogenous regeneration are trialed. This review briefly outlines the current state of knowledge on the molecular basis of the four modalities of myogenesis that have been identified in the developing vertebrate heart. Stem cell-mediated myogenic repair in the heart represents a fifth modality, and an exciting frontier with basic and practical implications that remain to be explored. PMID- 12757863 TI - Cardiac specific differentiation of mouse embryonic stem cells. AB - Embryonic stem (ES) cells may represent an alternative source of functionally intact cardiomyocytes for the causal treatment of cardiovascular diseases. However, this requires cardiac-specific differentiation of stem cells and the selection of pure lineages consisting of early embryonic cardiomyocytes. Therefore, an understanding of the basic mechanisms of heart development is essential for selective differentiation of embryonic stem cells into cardiac cells. The development of cardiac cells from embryonic stem cells is regulated by several soluble factors and signalling molecules together with cardiac specific transcription factors such as the zinc-finger GATA proteins and Nkx-2.5. GATA-4 and Nkx-2.5 seem to be essential for heart development. The use of enhanced green fluorescent protein (EGFP) under the control of cardiac-specific promoters in combination with the ES cell system has allowed for the functional characterisation of cardiac precursor cells. Embryonic stem cell-derived cardiomyocytes developmentally express similar cardiac-specific proteins, ion channels and signalling molecules to that of adult cardiomyocytes. Furthermore, identification of growth factors and signalling molecules under cell culture conditions is crucial for the selective cardiac differentiation of embryonic stem cells. Therefore, serum-free culture conditions have to be established in order to examine the influence of different growth factors and signalling molecules on cardiac development and/or formation from ES cells. Although significant progress has been made in generating cardiac cell lineage by the combination of genetically manipulative methods with selective culture conditions for cell transplantation therapy, one of the remaining future challenges for transplantation in humans is the immunological rejection of the engrafted cardiomyocytes. PMID- 12757864 TI - Twenty one years of P19 cells: what an embryonal carcinoma cell line taught us about cardiomyocyte differentiation. AB - Many different stem cell types have been shown to differentiate into cardiac muscle cells in vitro but P19 embryonal carcinoma (EC) cells were one of the first examples described and have been the most extensively characterized. P19 EC cells, isolated from an experimental embryo-derived teratocarcinoma in mice, are multipotent and can differentiate into cell types of all three germ layers. Because of their capacity to form cardiomyocytes however, they have been used to dissect the role of cardiac-specific transcription factors and upstream signalling pathways in cardiac cell differentiation. Furthermore, they have shed light on unique aspects of cardiac cell physiology during heart cell differentiation, including regulation of the proteins underlying the electrical and contractile systems. Here, we review studies on different subclones of P19 cells, and what they have taught us about cardiac differentiation and physiology. PMID- 12757865 TI - Development of heart muscle-cell diversity: a help or a hindrance for phenotyping embryonic stem cell-derived cardiomyocytes. AB - Despite the advances in cardiovascular treatment, cardiac disease remains a major cause of morbidity in all industrialized countries. The extraordinary potential of (embryonic) stem cells for therapeutic purposes has revolutionized ideas about cardiac repair of diseased cardiac muscle to exciting stages. This, in turn, has challenged research on cardiac differentiation of stem cells. For instance, cultures of mouse embryonic stem cells quite easily differentiate into the cardiogenic lineage, as assessed by their potential to beat spontaneously. However, repair of impaired cardiac muscle by spontaneously beating cardiac muscle cells might impose severe risks upon a human patient. Therefore, it is of crucial importance to understand the mechanisms that underlie the development of the distinct cardiac muscle cell types of the adult mammalian heart. In this review we tried to relate cardiac morphogenesis to the development of unique molecular phenotypes of cardiomyocytes. This relationship will provide a framework to assess the significance of the molecular phenotypes that are observed in embryonic stem cell-derived cardiomyocytes (ESDCs). Although for the phenotyping of ESDCs a comparison should be made with the phenotypes of the developing heart, so far none of the currently available markers allow unequivocal assignment of subtypes. PMID- 12757866 TI - Human embryonic stem cells for cardiovascular repair. AB - The critical loss of functional cardiomyocytes causes severe deterioration of pump function, resulting in heart failure. The possibility to regenerate or repair damaged or ischemic cardiac tissue is a great challenge for the future treatment of end-stage heart failure. As cardiomyocytes cannot be regenerated in adults, current therapeutic modalities for the treatment of end-stage heart failure are limited and include medical therapy, mechanical left ventricular assist devices, and cardiac transplantation. This review will focus on the potential use of human embryonic stem (hES) cell-derived cardiomyocytes and vascular cells, as a therapeutic tool for the treatment of myocardial infarction and end-stage heart failure. PMID- 12757867 TI - Origin and use of embryonic and adult stem cells in differentiation and tissue repair. AB - Stem cells are self-renewing, unspecialised cells that can give rise to multiple cell types of all tissues of the body. They can be derived from the embryo, foetus and adult. The ability of stem cells to divide but also to differentiate to specialised cell types like nerve and muscle, have made them candidates on which to base therapies for diseases and disorders for which no, or only partially effective, therapies are available. Replacement of defective or absent cells in defective tissues and organs could represent a cure. Here, we introduce the background to stem cell research and review the present state-of-the-art in stem cell biology, directed differentiation and tissue repair. In particular, we distinguish embryonic versus adult sources of stem cells and data derived from animal versus human experiments in order to place current research and perspectives for clinical application in their correct context. PMID- 12757868 TI - Myocyte and myogenic stem cell transplantation in the heart. AB - Cellular transplantation is emerging as a potential mechanism with which to augment myocyte number in diseased hearts. To date a number of cell types have been shown to successfully engraft into the myocardium, including fetal, neonatal, and embryonic stem cell-derived cardiomyocytes, skeletal myoblasts, and stem cells with apparent cardiomyogenic potential. Here we provide a review of studies wherein myocytes or stem cells with myogenic potential have been transplanted into the heart. In addition, issues pertaining to the tracking and functional consequences of cell transplantation are discussed. PMID- 12757869 TI - Skeletal muscle satellite cell transplantation. AB - Cell transplantation is currently gaining a growing interest as a potential new means of improving the prognosis of patients with cardiac failure. The basic assumption is that left ventricular dysfunction is primarily caused by the loss of a critical number of cardiomyocytes and that their replacement by new contractile cells could functionally 'regenerate' postinfarction scars in which these cells are implanted. Primarily for practical reasons, autologous skeletal myoblasts have been the first to undergo clinical trials but other cell types are also considered, particularly bone marrow stem cells which are attractive because of their autologous origin and their purported cardiomyocyte/endothelial transdifferentiation potential in response to cues present in the target organ. However, several key issues still need to be addressed including (1) the optimal type of donor cells, (2) the mechanism by which cell engraftment improves cardiac function, (3) the optimization of cell survival, and (4) the potential benefits of cell transplantation in nonischemic dilated cardiomyopathies. In parallel to the experimental studies designed to address these issues, clinical trials are underway and their results should hopefully allow the assessment of to what extent cellular therapy may improve the outcome of patients with heart failure. PMID- 12757870 TI - Cellular cardiomyoplasty--cardiomyocytes, skeletal myoblasts, or stem cells for regenerating myocardium and treatment of heart failure? PMID- 12757871 TI - Stem cells and cardiac disorders: an appraisal. AB - The use of stem cells has proved to be an important tool in investigating the events of early cardiac development, differentiation, and morphogenesis. In addition, stem cell transplantation in the treatment of certain cardiac disorders has shown early promise. We have attempted to present a balanced review of both basic studies and clinical-therapeutic potential of stem cells transplantation in the damaged heart. PMID- 12757872 TI - De novo vasculogenesis in the heart. AB - The formation of the embryonic heart vasculature is a complex process and is the result of vasculogenic, angiogenic and arteriogenic mechanisms, involving cells from distinct origins. In the neonate and the adult, several sources of endothelial precursor cells (EPCs) have been identified that contribute to physiological and pathological vascularization, consistent with the concept of de novo vasculogenesis after birth, including in the heart. The existence of EPCs in the adult has offered the possibility to use these cells for revascularization of ischemic tissues. An important challenge for vascular regeneration in ischemic and post-infarction patients is now to identify the most adequate cell source and cell dose for sufficient revascularization. This review gives an overview of the cellular and molecular cues involved in the formation of the heart vasculature before and after birth and discusses some of the recent insights and outstanding questions on EPCs and other vascular progenitors-both from a biological and therapeutic perspective. PMID- 12757873 TI - Post-natal endothelial progenitor cells for neovascularization in tissue regeneration. AB - The isolation of endothelial progenitor cells (EPCs) derived from bone marrow (BM) was an outstanding event in the recognition of 'de novo vessel formation' in adults occurring as physiological and pathological responses. The finding that EPCs home to sites of neovascularization and differentiate into endothelial cells (ECs) in situ is consistent with 'vasculogenesis', a critical paradigm well described for embryonic neovascularization, but proposed recently in adults in which a reservoir of stem or progenitor cells contributes to vascular organogenesis. EPCs have also been considered as therapeutic agents to supply the potent origin of neovascularization under pathological conditions. This review provides an update of EPC biology as well as highlighting their potential use for therapeutic regeneration. PMID- 12757874 TI - Cardiomyocytes derived from embryonic stem cells resemble cardiomyocytes of the embryonic heart tube. AB - OBJECTIVE: After formation of the linear heart tube a chamber-specific program of gene expression becomes active that underlies the formation of the chamber myocardium. To assess whether this program is recapitulated in in vitro differentiated embryonic stem cells, we performed qualitative and quantitative analyses of cardiogenesis in vivo and in vitro. METHODS: Gene expression profiles were made by in situ hybridisation and real-time PCR and electrophysiological profiles by patch clamp analyses of cardiomyocytes derived from time series of differentiating HM1 mouse embryonic stem cells and from embryonic and adult mouse hearts. RESULTS: In embryoid bodies the in situ patterns of expression of alpha myosin heavy chain, myosin light chain 2a and sarcoendoplasmic reticulum calcium ATPase 2a were similar to that of the heart muscle-specific marker gene cardiac troponin I. Myosin light chain 2v was expressed in part of the cardiac troponin I expressing area, indicating heterogeneity within the cardiac cell population. Atrial natriuretic factor expression, indicative of the chamber-type program, could only very occasionally be detected by in situ hybridisation. Quantitative reverse transcriptase PCR showed that all cardiac genes, most notably atrial natriuretic factor, were expressed at relatively low levels, similar to those in embryonic hearts at embryonic day 8.75-9. Analysis of the electrophysiological characteristics of embryonic stem cell-derived cardiomyocytes showed an increase of the upstroke velocity and a shorter duration of the action potential during prolonged differentiation in vitro. When embryonic mouse heart compartments of embryonic day 12.5 were used as a reference, the electrophysiological characteristics of a substantial part of the embryonic stem cell-derived cardiomyocytes were most reminiscent to those observed in the embryonic outflow tract. CONCLUSION: Together, these data suggest that most cardiomyocytes acquired by differentiation of embryonic stem cells maintain a phenotype reminiscent of that of the cardiomyocytes of the primary heart tube, and hardly any myocytes develop a chamber myocardial phenotype. PMID- 12757876 TI - The human adult cardiomyocyte phenotype. AB - AIM: Determination of the phenotype of adult human atrial and ventricular myocytes based on gene expression and morphology. METHODS: Atrial and ventricular cardiomyocytes were obtained from patients undergoing cardiac surgery using a modified isolation procedure. Myocytes were isolated and cultured with or without serum. The relative cell attachment promoting efficiency of several reagents was evaluated and compared. Morphological changes during long-term culture were assessed with phase contrast microscopy, morphometric analysis and immunocytochemistry or RT-PCR of sarcomeric markers including alpha-actinin, myosin light chain-2 (MLC-2) and the adhesion molecule, cadherin. RESULTS: The isolation method produced viable rod-shaped atrial (16.6+/-6.0%, mean+/-S.E.; n=5) and ventricular cells (22.4+/-8.0%, mean+/-S.E.; n=5) in addition to significant numbers of apoptotic and necrotic cells. Cell dedifferentiation was characterized by the loss of sarcomeric structure, condensation and extrusion of sarcomeric proteins. Cells cultured with low serum recovered and assumed a flattened, spread form with two distinct morphologies apparent. Type I cells were large, had extensive sarcolemmal spreading, with stress fibers and nascent myofibrils, whilst type II cells appeared smaller, with more mature myofibril organisation and focal adhesions. CONCLUSION: Characterization of the redifferentiation capabilities of cultured adult cardiac myocytes in culture, provides an important system for comparing cardiomyocytes differentiating from human stem cells and provides the basis for an in vitro transplantation model to study interaction and communication between primary adult and stem cell-derived cardiomyocytes. PMID- 12757875 TI - P19 embryonal carcinoma cells: a suitable model system for cardiac electrophysiological differentiation at the molecular and functional level. AB - OBJECTIVE: Murine P19 embryonal carcinoma (EC) cells can differentiate into spontaneously beating cardiomyocytes in vitro and have revealed important insight into the early molecular processes of cardiomyocyte differentiation. We assessed the suitability of the P19 cell model for studying cardiac ion channel regulation at the molecular and functional level. METHODS: P19 cells were induced to differentiate towards cardiomyocytes. mRNAs for cardiac markers and ion channels were determined by RT-PCR at six timepoints during the differentiation process. Action potentials and individual ion currents were measured by whole cell patch clamp. RESULTS: Ion channel mRNA expression of several channels is temporally regulated during differentiation, while others show little or no regulation. L type calcium and transient outward channels are expressed from very early on, while sodium and delayed and inward rectifier channels are upregulated at somewhat later stages during differentiation, which mirrors the in vivo murine cardiomyocyte differentiation during embryogenesis. Spontaneous cardiomyocyte action potentials exhibit a low upstroke velocity, which often can be enhanced by hyperpolarizing the cells, hence activating thusfar dormant ion channels to contribute to the action potential upstroke. Action potential duration decreases considerably during the differentiation of spontaneously beating cells. In late stages, non-beating myocytes can be found which only generate action potentials upon electrical stimulation. Their shape is comparable to neonatal/juvenile ventricular mouse myocytes in culture. Finally, we show that P19-derived cardiomyocytes display a very complete set of functional ion channels. CONCLUSION: P19 cells represent a powerful model to study the regulation of myocardial electrophysiological differentiation at the molecular and functional level. PMID- 12757877 TI - Survival and function of mouse embryonic stem cell-derived cardiomyocytes in ectopic transplants. AB - OBJECTIVE: Embryonic stem cell-derived cardiomyocytes are a useful source for cell transplantation into the heart, as well as for tissue engineering of the extracardiac vascular system. The present study was designed to investigate the survival and contractile function of embryonic stem cell-derived cardiomyocytes around large blood vessels to assess the feasibility of their ectopic use for future engineering of cardiovascular tissues. METHODS: The mouse embryonic stem cell-derived cardiomyocytes were transplanted into the retroperitoneum of the adult nude mice, and the myocardial tissues that developed were characterized by electrophysiological and histological techniques. RESULTS: Macroscopic and electrophysiological analyses showed spontaneously contracting transplants in the host retroperitoneum 7 and 30 days after transplantation. Immunohistochemistry detected developing cardiomyocytes in the transplants on Day 7, which formed the myocardial tissues. They were positive for cardiac troponin I, cadherin, connexin 43, and proliferating cell nuclear antigen, but negative for alpha-smooth muscle actin. Vascular formation was discernible in the transplant tissues. By Day 30, more mature myocardial tissues had been established in the transplants. Electron microscopic study emphasized that the transplant tissues comprised cardiomyocytes, in which myofibrils with organized sarcomeres were observed. Desmosomes, fasciae adherens and gap junctions were evident in the cellular junctions. CONCLUSIONS: The cardiomyocytes derived from the mouse ES cells were demonstrated to be viable and function in the ectopic site of the host retroperitoneum up to Day 30, following a process of proliferation and differentiation. Vascularization and host perfusion beneficial for the survival of the cardiomyocytes occurred in the transplants. PMID- 12757878 TI - Endoventricular porcine autologous myoblast transplantation can be successfully achieved with minor mechanical cell damage. AB - OBJECTIVE: Transplantation of skeletal myogenic precursor cells (mpc) into the myocardium using a non-surgical procedure. METHODS: Closed-chest mpc transplantation was assessed in pigs using the NOGA-Biosense device allowing both electromechanical mapping of the left ventricle (LV), and guided mpc injections through endocardium. RESULTS: We successively established that: (1) adequate preimplantation handling of mpc can be achieved when mpc are kept in 0.1% serum albumin-containing medium until implantation; (2) mpc are neither retained nor destroyed in the catheter or the needle and their passage does not affect their survival, growth and differentiation; (3) large numbers of autologous mpc can be actually transplanted in the LV myocardium by transendocardial route, as assessed by post-mortem examination of pigs injected with iron-loaded mpc; (4) cell injection into the myocardium does not induce conspicuous cell mortality since more than 80% of mpc recovered from LV tissue are alive 15 min after injection; (5) mpc injections can be guided into circumscribed LV targets such as infarcted areas, as assessed by comparison of map injection sites with location of iron loaded mpc at post-mortem examination of LV myocardium. CONCLUSION: This new approach may pave the way for a large spectrum of cell therapies targeting myocardial diseases. PMID- 12757879 TI - Temporal patterns of bone marrow cell differentiation following transplantation in doxorubicin-induced cardiomyopathy. AB - OBJECTIVE: Recent studies have suggested benefits of bone marrow cell transplantation for the regeneration of ischemic cardiac tissue. To extend the potential of cell transplantation, we assessed this treatment in a mouse model of acute nonischemic doxorubicin-induced cardiomyopathy. METHODS: To allow detection of engrafted cells, we used transgenic mice expressing the nuclear-located LacZ under the control of either desmin or vimentin promoters, which identify muscle lineage and mesenchymal cells, respectively. All transplanted cells were also labeled with the fluorescent dye DIL. One week after the administration of doxorubicin (15 mg/kg), mice were intramyocardially injected with either allogeneic unpurified bone marrow cells (6 x 10(6) in 30 microl, n=59) or purified sca-1(pos) cells (4 x 10(5) in 30 microl, n=22). In parallel, control normal mice received only unpurified bone marrow cells (n=28). Hearts were harvested at serial intervals until 2 weeks after transplantation and analyzed by immunohistochemistry to assess the degree of engraftment and transplanted cell differentiation. RESULTS: In control mice, no differentiation of bone marrow cells was detected. In contrast, unpurified bone marrow cells grafted into diseased myocardium featured two successive phases of cell differentiation. The first yielded cells with a mesenchymal phenotype (44.1+/-10.1 cells/3 x 10(-2) mm(3) at 2 days), was transient and lasted 1 week. The second phase was characterized by cells with a muscular phenotype detected in a small number of cells (5.6+/-2.3 cells/3 x 10(-2) mm(3) at 7 days). Two weeks after transplantation, some of these cells appeared phenotypically close to cardiomyocytes, as evidenced by morphology and positive staining for myosin binding protein C, vinculin and myosin heavy chain. In sca-1(pos) hematopoietic progenitor grafted mice hearts, no transdifferentiation into cardiac cells was detected at any time point. CONCLUSION: These data support the hypothesis of the potential for a myogenic differentiation of bone marrow cells following engraftment in a nonischemic model of global cardiomyopathy. Bone marrow-derived cells amenable to cardiac differentiation are present in total unpurified bone marrow but not in the sca-1(pos) hematopoietic progenitor cell population. However, the very small number of transdifferentiated cells raises concerns over their functional efficacy. PMID- 12757880 TI - Growth and differentiation of rat bone marrow stromal cells: does 5-azacytidine trigger their cardiomyogenic differentiation? AB - OBJECTIVE: The potential use of bone marrow stromal cells (MSCs) as a cellular therapy for chronic cardiac diseases relies on the ability of the cell to replicate extensively in vitro and to give rise to myogenic cells that can replace the damaged cardiomyocytes. For this reason the present study investigated the replication lifespan and chemical-induced cardiomyogenic differentiation of rat MSCs in vitro. METHODS: The primary and the successively passaged Wistar rat MSCs were exposed to different concentrations (3, 5 and 10 microM) of 5-azacytidine using different methods (single- or repeat-treatment). The growth properties and the fate of the cells were compared to their untreated counterparts by cell counting, immunocytochemistry and Western analysis. RESULTS: When seeded at a density of 2845 cells/cm(2) and cultured under common conditions, rat MSCs could be expanded up to 21.94 cell doublings in 30 days of successive subcultures. This was accompanied by a gradual loss of their replication ability with passages. When treated with 5-azacytidine for 24 h at day 3 of primary culture and the first subculture, the growth properties of the MSCs were not obviously affected. Neither the spontaneously beating cells nor the formation of myotubes were found in the primary and first passaged MSCs after a single treatment with 5-azacytidine and in cultures which underwent repeated 5 azacytidine-treatments during continuous subculturing to passage 2. The expressions of cardiac troponin I, cardiac myosin heavy chain and connexin 43 by the 5-azacytidine-treated MSCs were also undetectable at both immunocytochemistry and Western blot levels. The specificity and reliability of the detection methods were technically confirmed with cultured rat cardiomyocytes. CONCLUSIONS: Rat MSCs cannot be extensively expanded in vitro or be induced to differentiate in an expected cardiomyogenic way by 5-azacytidine-treatment, if the cells are not immortalized. PMID- 12757882 TI - Endothelial progenitor cell culture and differentiation in vitro: a methodological comparison using human umbilical cord blood. AB - OBJECTIVE: Endothelial progenitor cells (EPC) can contribute to vascular repair and targeted tumour therapy. Little is known about generating EPC from human umbilical cord blood. We therefore compared methods for purification of EPC from human umbilical cord blood. METHODS: Mononuclear cells were isolated from human umbilical cord blood by density gradient centrifugation and used either unselected or after CD34 preselection. Unselected mononuclear cells were cultured for 9 days. Culture-dish-adherent (CDAC) and non-adherent (CDNAC) CD34+ cells were cultured separately for 4 weeks. Surface markers were assessed by immunofluorescence staining and FACS analysis. RESULTS: In unselected mononuclear cells, VEGF-R2 and VE-cadherin expression increased up to day 6. They stained positive with UEA-1 and took up acetylated LDL. Expression of CD45 and CD14 decreased over time, but remained strong. CD133 and CD34 were not expressed. CD34+-CDNAC acquired an endothelial phenotype over time with an increase of VEGFR 2 and von Willebrand factor (vWF). CD45 and CD14 decreased, while CD34 and the progenitor-cell marker CD133 remained strongly expressed. CD34(+)-CDAC showed a strong increase in VEGFR-2, CD133, CD34 and vWF, while CD14 decreased, and CD45 did not change. CONCLUSION: Putative EPC can be obtained from human umbilical cord blood. When selected for CD34, cells can be differentiated in culture to express markers of mature endothelial cells, while keeping progenitor markers. In contrast, short-term culture of unselected mononuclear cells leads to an endothelioid-monocytoid phenotype devoid of progenitor markers. Thus, the outgrowth from CD34-selected cells appears to be superior to short-term culture of unselected mononuclear cells with regard to endothelial cell-lineage specific differentiation of cells with a progenitor marker profile. PMID- 12757881 TI - Vascular gene delivery of anticoagulants by transplantation of retrovirally transduced endothelial progenitor cells. AB - OBJECTIVE: Recent studies have documented the presence of bone marrow-derived endothelial progenitor cells (EPC) in the circulation of several species. This study was designed to evaluate the use of engineered EPC for vascular gene delivery into angioplasty-induced arterial lesions. METHODS AND RESULTS: EPC could easily be isolated from whole bone marrow and peripheral blood of adult rats. Differentiation was induced by culture on fibronectin in the presence of endothelial specific growth factors. Rat EPC shared several phenotypic and functional properties with mature endothelial cells. Recombinant retroviruses were generated encoding for the anticoagulants tissue-type plasminogen activator (tPA) and hirudin. Efficient (>90%) ex vivo gene transfer could be achieved resulting in high levels of transgene production. Engineered EPC were locally infused into freshly balloon-injured carotid arteries. Analysis of day 7 vessels showed 73+/-10% luminal coverage of the lesioned arterial bed with transduced EPC. Sustained secretion of both anticoagulants could be detected in organ cultures of explanted arteries. EPC seeding inhibited dilation of the injured arterial segment and prevented reduction of media thickness. However, rapid repopulation with EPC failed to attenuate neointima formation in this model. CONCLUSIONS: Peripheral blood and bone marrow can be used as source for endothelial lineage cells. Cultured EPC can be genetically engineered by retroviral gene transfer and serve as cellular vehicles for vascular gene and drug delivery of anticoagulants. Local transplantation of EPC attenuates reendothelialization of angioplasty-injured arteries but fails to inhibit neointima proliferation. PMID- 12757883 TI - Neoendothelialization after peripheral blood stem cell transplantation in humans: a case report of a Tokaimura nuclear accident victim. AB - OBJECTIVES: Neoendothelialization by circulating endothelial progenitor cells has been a topic of recent research. The extent and scale of this process in humans is not well understood. We examined the extent of neoendothelialization of the aorta and peripheral arteries in the case of a patient who underwent peripheral blood stem cell transplantation for acute radiation syndrome. METHODS: Human tissue samples from the aorta and peripheral arteries were obtained at autopsy. Endothelial cells were isolated, confirmed by von Willebrand factor immunostaining, and then subjected to fluorescent in situ hybridization analysis using X- and Y-chromosome specific probes to examine neoendothelialization by donor cells as possible in this case in which the donor and recipient were of different genders. RESULTS: The aorta showed almost 25% of all endothelial cells to be replaced by donor-origin endothelial cells. The peripheral arteries were also replaced but to a lesser extent. DISCUSSION: The present study provides evidence that peripheral blood is a source of endothelial progenitor cells in humans. Neoendothelialization of the aorta occurs to a significant extent under certain conditions suggesting the potential for exploitation of therapeutic neovascularization by transplantation of circulating endothelial progenitor cells. PMID- 12757884 TI - Interleukin-1beta increases spinal cord wind-up activity in normal but not in monoarthritic rats. AB - Cytokines produced by spinal cord glia after peripheral inflammation, infection or trauma have a relevant role in the maintenance of pain states. The effect of intrathecally administered interleukin-1beta (IL-1beta) on spinal cord nociceptive transmission was studied in normal and monoarthritic rats by assessing wind-up activity in a C-fiber-mediated reflex paradigm evoked by repetitive (1 Hz) electric stimulation. Low i.t. doses of IL-1beta (0.03, 0.12, 0.5 and 2.0 ng) dose-dependently enhanced wind-up activity in normal rats, while higher doses (8.0 ng) only produced a marginal unsignificant effect. IL-1beta administration to monoarthritic rats did not significantly change wind-up scores at any dose. Adaptive changes developed in the spinal cord during chronic pain may underlie the ineffectiveness of exogenous IL-1beta to up-regulate nociceptive transmission. PMID- 12757885 TI - Brain regions sensitive to the face inversion effect: a functional magnetic resonance imaging study in humans. AB - Perception of upright faces relies on configural processing. Therefore recognition of inverted, compared to upright faces is impaired. In a functional magnetic resonance imaging experiment we investigated the neural correlate of a face inversion task. Thirteen healthy subjects were presented with a equal number of upright and inverted faces alternating with a low level baseline with an upright and inverted picture of an abstract symbol. Brain activation was calculated for upright minus inverted faces. For this differential contrast, we found a signal change in the right superior temporal sulcus and right insula. Configural properties are processed in a network comprising right superior temporal and insular cortex. PMID- 12757886 TI - Continuity of the processing of declarative knowledge during human sleep: evidence from interrelated contents of mental sleep experiences. AB - The positive influence of sleep on memory may partly depend on the processing which transforms items of declarative knowledge into contents of mental sleep experience (MSE). This view implies that the consolidation level should be more enhanced for those items which have been repeatedly processed and transformed into identical or very similar (so-called interrelated) contents of distinct MSEs in the same night. We examined here the occurrence of interrelated contents in the MSEs reported after an awakening provoked in stage 2 at sleep onset and the spontaneous awakening in the morning. Interrelated contents resulted much more frequently than the chance occurrence of contents with the same semantic features, regardless of the sleep stage in which morning awakening occurred. The accessibility of given items for transformation into MSE contents over the night makes it plausible that they are reprocessed, and thus further consolidated, during various stages and cycles of sleep. PMID- 12757887 TI - Immunohistochemical study on the distribution of alpha-synuclein in the central nervous system of transgenic mice expressing a human Cu/Zn superoxide dismutase mutation. AB - We used the SOD1(G93A) transgenic mice as an in vivo model of amyotrophic lateral sclerosis (ALS) and performed immunohistochemical studies to investigate whether alpha-synuclein is involved in the pathogenesis of ALS. In the spinal cord of transgenic mice, immunohistochemistry showed intense staining of alpha-synuclein mainly in the anterior horn. In the hippocampus of transgenic mice, differential increases in the staining density of alpha-synuclein were observed. In the cerebellar cortex of transgenic mice, the prominent immunostaining of alpha synuclein was found in the molecular and granular layers. The present study provides the first in vivo evidence that alpha-synuclein immunoreactivity was increased in the central nervous system of SOD(G93A) transgenic mice, suggesting that alpha-synuclein might play an important role in the pathogenesis of ALS. However, the functional implications of these increases require elucidation. PMID- 12757888 TI - Plasma amyloid beta protein 1-42 levels are increased in old Down Syndrome but not in young Down Syndrome. AB - Plasma amyloid beta protein 1-40 (Abeta40) and Abeta42 levels were quantitated from 28 young Down syndrome (DS) (20-40 years old), 28 age-matched controls, 32 old DS (41-65 years old) and 32 age-matched controls in a sandwich enzyme-linked immunosorbent assay. Abeta40 levels were higher in young DS and old DS than controls. Abeta42 levels in young DS and controls were similar, however Abeta42 levels were higher in old DS than controls or young DS. The higher Abeta42 levels in old DS suggests that Abeta42 is selectively increased in plasma concurrently with the development of Alzheimer disease neuropathology. PMID- 12757889 TI - Overexpression of beta-1,4-galactosyltransferase I in rat Schwann cells promotes the growth of co-cultured dorsal root ganglia. AB - The cell surface beta-1,4-galactosyltransferase I (beta-1,4-GalT-I) functions as one of the receptors of laminin during the neurite outgrowth on basal lamina by binding to N-linked oligosaccharides in the laminin E8 domain. In this study, we demonstrated that the purified rat Schwann cells transfected with the expression plasmid of beta-1,4-GalT-I cDNA transiently promoted outgrowth and elongation of the neurites from co-cultured rat dorsal root ganglia, while those transfected with the antisense expression plasmid of beta-1,4-GalT-I had the opposite effects. These results suggested that the expression of beta-1,4-GalT-I in Schwann cells of peripheral nerve might promote both growth of developmental neuron and regeneration of injured nerve. PMID- 12757890 TI - Differentiating primary central nervous system lymphoma from glioma in humans using localised proton magnetic resonance spectroscopy. AB - In order to characterise primary central nervous system lymphomas (PCNSL) and to evaluate if 1H spectroscopy improves the preoperative differential diagnosis of PCNSL and glioma, seven immunocompetent patients with PCNSL and 21 patients with glioma were examined using single voxel, short echo time magnetic resonance spectroscopy (MRS; 1.5 T, STEAM 1500/20). All PCNSL demonstrated massively elevated lipid resonances and markedly elevated choline. Similarly increased lipid resonances were only found in seven necrotic glioblastomas, PCNSL differing from all solid astrocytomas by massively elevated lipid resonances. Additionally, PCNSL had higher Cho/Cr ratios than all grades of astrocytoma. In conclusion, we found that massively elevated lipid resonances are a hallmark of PCNSL in immunocompetent patients. Together with a markedly elevated Cho/Cr ratio, MRS provides metabolic information which may improve the preoperative differentiation of PCNSL and glioma. PMID- 12757891 TI - The correspondence between the labeling patterns of antibody RT97, neurofilaments, microtubule associated protein 1B and tau varies with cell types and development stages of chicken retina. AB - The correspondence between the labeling patterns of antibody RT97, neurofilaments (NF-M and NF-H), microtubule associated protein 1B (MAP1B) and tau, were studied in the developing chicken. At embryonic day 3 (E3), intense RT97 immunoreactivity (IR) was found to be localized in cells in the region adjacent to the intraretinal space, which separates the inner and outer layers of the optic cup, and this was sustained at E8. However, this pattern changed dramatically at E12, as the intensities of RT97 IR increased in the inner retinal layer, while the outermost layers showed only weak IR. The adult stage retina showed RT97 IR within the nerve fibers of the ganglion cells, the processes of the amacrine cells and the photoreceptors. Additional immunostainings for NF-M, -H, MAP1B and tau showed that the observed changes in RT97 IRs were due to the different expressions of these proteins at different development stages. PMID- 12757892 TI - Relational discrimination learning between amplitude-modulated sounds in the rat. AB - We studied the cue used by rats to discriminate between amplitude-modulated (AM) sounds. Water-deprived rats were exposed to two sounds, and licking a spout during the presentation of one of the two (S+) was rewarded with water, while licking during the other (S-) was not. The AM depth was 100, 60 or 20%. Rats were trained to discriminate between 100 and 60% AM sounds over 2 consecutive days. On the next day, the performance in discriminating between 60 and 20% AM sounds was evaluated. Of the four possible combinations of stimuli, a good performance was observed only when the relationship between S+ and S- regarding the AM depth was kept constant throughout the experiments, indicating that the relationship regarding the AM depth was the cue used by the rats for the discrimination. PMID- 12757893 TI - Novel variants of murine serotonin transporter mRNA and the promoter activity of its upstream site. AB - Three variants of murine serotonin transporter (5-HTT) mRNA, which consist of a different exon-one (exon 1a, exon 1b or exon 1c) and the same exon-two to exon five, were identified. The promoter region for each exon 1 (p1a, p1b and p1c, respectively), ligated to pGL-3 enhancer vector, had activities significantly higher than the empty vector in all cell lines tested except p1c in PC-12, whereas the activity of p1c was significantly lower than the others. Effects of the treatment of dibutyryl-cyclic AMP, human interferon-alpha or mouse interferon gamma have different profiles among COS-7, PC-12, C6 glioma and immortalized rat serotonergic raphe neurons, RN46A. These three promoter regions may play a role in the transcription of the 5-HTT and could offer a model of the regulation of 5 HTT production in humans and further the pathogenesis of depression. PMID- 12757894 TI - The serotonergic modulation of synaptic plasticity in the rat hippocampo-medial prefrontal cortex pathway. AB - The ability of the serotonergic (5-HTergic) system to affect the hippocampo medial prefrontal cortex (mPFC) synaptic properties was examined in rats with lesions of 5-HTergic neurons. Intracerebroventricular injections of 5,7 dihydroxytryptamine (5,7-DHT) resulted in selective depletion of 5-HT and 5 hydroxyindoleacetic acid in the cerebral cortex, hippocampus and raphe regions. The 5,7-DHT-lesioned rat had no changes in basal synaptic transmission in the hippocampo-mPFC pathway. Conversely, we observed the augmentation of short-term synaptic plasticity, i.e. paired-pulse facilitation, when compared with sham operated rats in this pathway. The magnitude of long-term potentiation (LTP) was significantly augmented in 5,7-DHT-lesioned rats. This augmentation of hippocampo mPFC LTP had a significant correlation with the degree of cortical 5-HT levels. These results suggest that the 5-HTergic system may modulate plastic properties at the hippocampal-mPFC synapses in vivo. PMID- 12757895 TI - Growth hormone secretagogue receptor expression in the cells of the stomach projected afferent nerve in the rat nodose ganglion. AB - Growth hormone secretagogue receptor (GHS-R) is widely expressed in various regions of the body, such as the brain, pituitary gland, heart and gastrointestinal tract. Recently, ghrelin, an endogenous ligand for GHS-R, was found in the rat stomach, and several studies have suggested that ghrelin acts via the vagal afferent nerve. In this study, we studied the expression of GHS-R mRNA in the rat nodose ganglion by reverse transcriptase-polymerase chain reaction and in situ hybridization, the results of which clearly demonstrated the presence of GHS-R mRNA and GHS-R producing cells in the rat nodose ganglion. We also studied the retrograde tracing of nodose ganglion cells to the stomach and found that some GHS-R mRNA-expressing cells contain the retrograde rebelling. Our results provide direct morphological evidence that GHS-R is produced in afferent neurons of the nodose ganglion and suggest that ghrelin signals from the stomach are transmitted to the brain via vagal afferent nerves. PMID- 12757896 TI - Central mechanisms for two-point discrimination in humans. AB - We studied the cognitive mechanisms for two-point discrimination (TPD) in 11 normal subjects, using electrical pulses. We used six ball-shaped electrodes placed in line on the dorsal surface of the left hand, and two-point was stimulated by two electrodes randomly selected. We measured the reaction time for TPD and calculated the percentage of correct responses for each two-point stimulation. The subjects' response was significantly affected by the preceding stimuli as well as the distance of the stimuli: for a two-point stimulus condition, subjects tended to feel the stimuli as two-point when the distance between the stimuli was longer than that of preceding stimuli, whereas they felt the stimuli as one-point when the distance was shorter than that of the preceding stimuli. The present results indicate that the TPD process involved evaluation of the distance between the stimuli relatively to that of the preceding stimuli, as well as evaluation of absolute distance between the stimuli. PMID- 12757897 TI - Steady-state visual evoked potentials reveal frontally-mediated working memory activity in humans. AB - Steady-state visual evoked potentials (SSVEPs) reflect power changes at the stimulus driving frequency and have been used to assess brain activity reflecting cognitive processing. Only one study has demonstrated SSVEP modulation associated with working memory (WM), and none have compared the spatial localization of SSVEP modulations during WM performance with other brain imaging methods. Here we examined WM-related activity recorded with dense-array SSVEPs, analyzed using low resolution electromagnetic tomography, and compared the results to our previous findings using functional magnetic resonance imaging (fMRI). WM was associated with increased SSVEP activity over the right dorsolateral prefrontal cortex, paralleling our previous fMRI findings. Frontal WM-related SSVEP power correlated selectively with task performance. These results demonstrate the utility of SSVEPs for studying representational aspects of cognition. PMID- 12757898 TI - Rapid eye-movement sleep deprivation does not 'rescue' developmentally regulated long-term potentiation in visual cortex of mature rats. AB - The age at which it is possible to obtain a usually age-limited (developmental) form of long-term potentiation (LTP) in rat visual cortex slices can be extended by suppressing rapid eye movement (REM) sleep. In this study, we examined whether REM sleep deprivation can also 'rescue' this type of LTP in older rats. Rats, 42 59 days of age, were either REM sleep-deprived for 7-10 days (n=8), or not deprived of REM sleep (control group, n=8). Brain slices from visual cortex were tested for the developmental form- and a related, non-developmental form of LTP. Three of the eight REM sleep-deprived animals and four of the eight non-deprived animals met criteria for a valid attempt to induce the developmental form of LTP. Though the non-age-regulated form of LTP could be obtained in all seven of these animals, the developmental form could not be elicited in any, indicating that REM sleep deprivation does not uniformly affect all forms of LTP in adult rats. We conclude that extended periods of REM sleep deprivation do not facilitate induction of developmentally regulated LTP once the animal is beyond a certain age. PMID- 12757899 TI - Estradiol reduces seizure-induced hippocampal injury in ovariectomized female but not in male rats. AB - Estrogens protect ovariectomized rats from hippocampal injury induced by kainic acid-induced status epilepticus (SE). We compared the effects of 17beta-estradiol in adult male and ovariectomized female rats subjected to lithium-pilocarpine induced SE. Rats received subcutaneous injections of 17beta-estradiol (2 microg/rat) or oil once daily for four consecutive days. SE was induced 20 h following the second injection and terminated 3 h later. The extent of silver stained CA3 and CA1 hippocampal neurons was evaluated 2 days after SE. 17beta Estradiol did not alter the onset of first clonus in ovariectomized rats but accelerated it in males. 17beta-Estradiol reduced the argyrophilic neurons in the CA1 and CA3-C sectors of ovariectomized rats. In males, estradiol increased the total damage score. These findings suggest that the effects of estradiol on seizure threshold and damage may be altered by sex-related differences in the hormonal environment. PMID- 12757900 TI - Modulation of the human glucocorticoid receptor function by antidepressive compounds. AB - Hyperactivity of the hypothalamic-pituitary-adrenal axis is associated with depression. We investigated the effect of various types of antidepressant agents in vitro on the function of glucocorticoid receptor (GR). Desipramine, clomiplamine, fluoxetine, milnacipran and clorgyline all induced rapid and sustained translocation of GR into the nucleus of human lymphocytes. In contrast, major and minor tranquilizers, lithium and verapamil, a blocker of membrane steroid transporter, were without effect. These antidepressants did not affect GR mediated transcription by themselves, but significantly inhibited dexamethasone induced transcription. These results indicate that structurally different antidepressants induce translocation of GR and inhibit GR-mediated transcription. PMID- 12757901 TI - Amygdala damage impairs emotion recognition from scenes only when they contain facial expressions. AB - Bilateral damage to the human amygdala impairs recognition of negatively valenced emotions from facial expressions, but it is unclear if this finding generalizes to richer visual stimuli that contain cues in addition to faces. We investigated this issue in 4 subjects with bilateral amygdala damage, 23 with unilateral amygdala damage, 22 brain-damaged controls and 16 normal individuals. Subjects were shown two blocks of complex social scenes; all stimuli in the two blocks were identical, except that the first block had all facial expressions in the image erased. While control subjects were more accurate in recognizing emotions when facial expressions were present, subjects with bilateral amygdala damage did not show the same benefit for negative emotions, often performing equivalently across the two conditions. Most striking, subjects with bilateral amygdala damage were more accurate in recognizing scenes showing anger with faces erased than with faces present, an effect resulting in part from highly abnormal recognition of certain angry facial expressions. All four subjects with bilateral amygdala damage were impaired in recognizing angry faces shown in isolation, and frequently mistook expressions of anger for smiles, a mistake never made by any control subject. Bilateral amygdala damage thus disproportionately impairs recognition of certain emotions from complex visual stimuli when subjects utilize information from facial expressions. PMID- 12757903 TI - Spatial and temporal processing in a subject with cortical blindness following occipital surgery. AB - Blindsight subjects are typically better at discriminating rapid, transient visual events than those with gradual on/off-sets. Surprisingly, the detailed investigation of temporal characteristics of mechanisms mediating blindsight is only reported in one subject (GY). It is of interest to establish whether these characteristics are similar to those in other cases of blindsight. Here, we report on a systematic study of spatio-temporal properties of mechanisms mediating blindsight in a subject VN. VN has a lower right quadranopia following surgical removal of the left occipital cortex above the calcarine sulcus, therefore, there are no remaining islands of intact visual cortex within this area. Similar to GY, the blindsight mechanisms in VN have narrowly tuned band pass temporal characteristics with a peak sensitivity at 20Hz and above chance performance at temporal frequencies >/=10 and 4.8->7.2 log of all the viruses tested, i.e. Sindbis, HSV-1, vaccinia, polio-1, EMC, HAV, CPV, BPV and SV40. However, in the presence of protein, i.e. 5% v/v foetal-calf serum (0.2% w/v protein), virus inactivation was less effective. At a dose of 2.0 J/cm2, virus inactivation was 5.0->6.4 log, however, HSV-1 (3.8 log), BPV (2.4 log) and SV40 (2.9 log) were all relatively resistant. This virus inactivation procedure may have application for increasing the safety of therapeutic biological products. PMID- 12757922 TI - A rapid method for immunotitration of influenza viruses using flow cytometry. AB - Reliable assays for accurate titration of influenza virus in infectious samples are pivotal to both influenza research and vaccine development. A titration assay adopted commonly for this purpose is the plaque assay on Madin-Darby canine kidney (MDCK) cells, despite it being time and labour consuming. A novel assay is described for titration of influenza viruses based on the detection of intracellular viral nucleoprotein (NP) by fluorescence-activated cell sorting (FACS). By using a panel of viruses of different type, subtype and origin, it is demonstrated that there is a mathematical correlation between titres measured by immunotitration and by classical plaque assay on MDCK cells. Moreover, the availability of NP antibodies specific for type A or type B influenza virus ensures the specificity of the assay. Based on speed, accuracy and specificity, it is concluded that the FACS-based immunotitration of influenza virus represents a valid and efficient alternative to the classical plaque assay. PMID- 12757923 TI - A real-time TaqMan PCR for routine quantitation of cytomegalovirus DNA in crude leukocyte lysates from stem cell transplant patients. AB - A real-time TaqMan PCR based on the cytomegalovirus (CMV) polymerase (pol) gene was developed for quantitation of CMV DNA in crude peripheral blood leukocyte (PBL) lysate from stem cell transplantation (SCT) patients. The dynamic range of the assay was between 10 and 4x10(6) copies. Both intra- and inter-assay variability were well within +/-0.25 log10 S.D. Thus, a pooled PBL sample that was used as positive control in 57 consecutive TaqMan PCR runs over 7 months showed a stable CMV quantity (4.12+/-0.13, log10 mean+/-S.D.). The sensitivity of the pol TaqMan PCR was validated by parallel analysis of 177 PBL samples with a nested PCR. The use of crude PBL lysate as PCR input did not cause PCR inhibition. We demonstrated further the clinical utility of the newly developed TaqMan PCR by monitoring changes in CMV levels in eight patients receiving antiviral therapy. This TaqMan PCR was highly sensitive, reproducible, and stable and has served a useful tool for monitoring CMV DNA levels in large number of clinical samples in a routine diagnostic setting for over 1 year. PMID- 12757924 TI - A new one-step RT-PCR method for virus quantitation in murine AIDS. AB - The causative agent of murine AIDS (MAIDS) in C57BL/6 mice, is a defective murine leukemia virus (BM5d) that requires the replication-competent helper virus (BM5e). Since this animal model of immunodeficiency, which shows many similarities to human AIDS, is also used to test the efficacy and toxicity of antiretroviral drugs, a method that allows the quantitative detection of both viruses would be very useful also if hampered potentially by endogenous viral sequences usually present in mice. While BM5d alone could induce the disease, the effect of BM5e on the immune system of diseased mice is unclear. A specific and reliable one-step RT-PCR method was developed for the co-amplification, with the same efficiency, of BM5d or BM5e with ss-actin used as an internal standard. The standard curves produced with cloned cDNA sequences (ss-actin and BM5d or BM5e) assure that all samples are analyzed during the exponential phase of the reaction. Using this new assay which provided a dynamic range of at least four log-unit, the ratio of initial absolute amounts of the virus and ss-actin RNA was determined, obtaining quantitative information on virus-specific cellular transcript in the lymph nodes and spleen during the natural history of the disease and during therapeutic regimens. PMID- 12757926 TI - Detection of specific reticuloendotheliosis virus sequence and protein from REV integrated fowlpox virus strains. AB - The detection is described of reticuloendotheliosis virus (REV) protein in tissue culture of chicken embryonated cells (CEFs) infected with field isolates of fowl poxvirus (FPV). By the polymerase chain reaction (PCR), five out of the six field isolates, but two out of the seven vaccine strains of FPV, were found to have had a 291 bp repeat sequence of REV-LTR integrated in their genomic DNA. An immunofluorescence (IF) method was employed using a monoclonal antibody (MAb) known to specify strain common envelope proteins for REV and allowed to detect the presence of a specific REV protein. The IF results indicate the localization of REV proteins in boundaries defined precisely within cells infected with these field strains of FPV carrying REV (FPV-REV). Furthermore, by immunoblotting (IB) using a chemiluminescent detection kit, the REV protein reacted specifically with the MAb and had a relative molecular mass (RMM) of 62 kDa. The data have the potential to advance substantially the current understanding of the integrated REV in FPV strains; and the identification of a unique protein associated with variant forms of FPV will also offer great potential for identification of novel vaccine candidates for use in poultry against variant forms of FPV. PMID- 12757925 TI - Production of recombinant Potato mop-top virus coat protein in Escherichia coli and generation of antisera recognising native virus protein. AB - Potato mop-top virus (PMTV, Pomovirus) is difficult to detect because it is unevenly distributed and present at low concentration in infected tissues. The production of PMTV-free seed relies on sensitive and specific detection methods of virus detection, including serological methods. The possibility of using a PMTV recombinant coat protein (CP) as an antigen for antiserum production was investigated. The region encoding the PMTV CP was inserted into pET3A, expressed in Escherichia coli, and the recombinant PMTV CP produced was used to raise antibodies in rabbits. Three antisera were produced. All recognised efficiently the recombinant CP in Western blot analysis and the most sensitive antiserum (H5003) detected native CP on Western blots and in ELISA. Thus, recombinant CP can be used as an alternative to purified virus for the production of specific antibodies against PMTV. PMID- 12757927 TI - Tagging of viral RNA transcripts with strain-specific oligonucleotides: characterization and application. AB - Binding('tagging') of a virus-specific oligonucleotide 'sticker' to RNA transcripts copied from PCR products caused retardation of transcript mobility in gel. This enables detection of specific sequences within the RNA transcripts, and a virus strain (PVY(NTN)) could thus be positively identified. We have demonstrated further that oligonucleotides that contained virus sequences originated from different genomic locations varied in their inhibitory effect on the rate of transcript migration in gel; thus, the most effective oligonucleotide could be chosen. Combinations of different strain-specific oligonucleotides had additive retarding effects on transcript migration. The conditions for annealing oligonucleotides to the RNA transcripts were studied, including concentrations of oligonucleotides and salt. A higher electrophoresis temperature (up to 45 degrees C) reduced the gel retardation phenomena, which indicated a conformation mechanism. The applicability of 'tagging' of RNA transcripts with a strain specific oligonucleotide for virus strain differentiation is discussed. PMID- 12757928 TI - Generation of recombinant influenza virus using baculovirus delivery vector. AB - A recombinant baculovirus vector containing mammalian cell-active promoters and transcription terminators was used to deliver a mutated influenza NS gene into Vero cells. In addition to the influenza NS gene, the baculovirus contained a reporter gene expression cassette (Green fluorescent protein, GFP), allowing to monitor the Vero cell transduction efficiency. More than 90% of Vero cells were expressing GFP 24-48 h post transduction. After infecting baculovirus transduced cells with influenza helper virus, progeny of attenuated influenza virus carrying the recombinant NS gene could be selected. Baculovirus delivery was highly reproducible and efficient in Vero cells. This new method for influenza gene delivery could contribute to influenza virus research and vaccine development. PMID- 12757929 TI - Physiological and pathophysiological functions of the ecto-nucleotide pyrophosphatase/phosphodiesterase family. AB - The ecto-nucleotide pyrophosphatase/phosphodiesterase (E-NPP) multigene family contains five members. NPP1-3 are type II transmembrane metalloenzymes characterized by a similar modular structure composed of a short intracellular domain, a single transmembrane domain and an extracellular domain containing a conserved catalytic site. The short intracellular domain of NPP1 has a basolateral membrane-targeting signal while NPP3 is targeted to the apical surface of polarized cells. NPP4-5 detected by database searches have a predicted type I membrane orientation but have not yet been functionally characterized. E NPPs have been detected in almost all tissues often confined to specific substructures or cell types. In some cell types, NPP1 expression is constitutive or can be induced by TGF-beta and glucocorticoids, but the signal transduction pathways that control expression are poorly documented. NPP1-3 have a broad substrate specificity which may reflect their role in a host of physiological and biochemical processes including bone mineralization, calcification of ligaments and joint capsules, modulation of purinergic receptor signalling, nucleotide recycling, and cell motility. Abnormal NPP expression is involved in pathological mineralization, crystal depositions in joints, invasion and metastasis of cancer cells, and type 2 diabetes. In this review we summarize the present knowledge on the structure and the physiological and biochemical functions of E-NPP and their contribution to the pathogenesis of diseases. PMID- 12757930 TI - Expression of mutant and wild-type TIMP3 in primary gingival fibroblasts from Sorsby's fundus dystrophy patients. AB - Gingival fibroblast cell lines were derived from Sorsby's fundus dystrophy (SFD) patients carrying the S181C TIMP3 and the E139X TIMP3 mutations. These cell lines were grown in culture to study expression of the wild-type and mutant tissue inhibitor of metalloproteinase 3 (TIMP3) alleles from a normal diploid cell type. Firstly, patient cells were found to co-express the wild-type and mutant TIMP3 alleles, S181C TIMP3 or E139X TIMP3, at the mRNA level using restriction fragment length polymorphism (RFLP) analysis. A SpeI RFLP for E139X TIMP3 is described. Low levels of endogenous TIMP3 protein expression were elevated using the natural polysaccharide calcium pentosan polysulfate (CaPPs) in combination with the cytokine IL-1alpha. Immunoblotting detected protein expression from both wild type and mutant alleles, S181C TIMP3 or E139X TIMP3. S181C TIMP3 from these cells was found to dimerise and retain MMP2 inhibitory activity. To facilitate studies of the E139X TIMP3 protein, the allele was expressed using HighFive insect cells. In this cell type, the E139X TIMP3 was synthesised as a mixture of monomer and dimer. Both monomeric and dimeric E139X TIMP3 protein retained MMP2 inhibitory activity in gelatin zymography. Expression of mutant E139X or S181C TIMP3 protein from a normal diploid patient-derived fibroblast cell had no effect on either MMP2 or MMP9 expression or activation whilst transcribed from their normal promoter context. PMID- 12757931 TI - Betaine homocysteine methyltransferase: gene cloning and expression analysis in rat liver cirrhosis. AB - It has been known for over half a century that homocysteine levels are elevated in liver cirrhosis, but the basis for it is not fully understood. Using differential display, we identified betaine homocysteine methyltransferase (BHMT) as a gene down-regulated in rat liver cirrhosis and most likely involved in this dysregulation. A partial BHMT clone was isolated by screening of a cDNA library with the differential display fragment. The full-length gene was generated by primer extension of cDNA. Expression levels of BHMT in cirrhotic livers of bile duct ligated rats were compared to controls by Northern and Western blotting as well as by enzyme activity measurements. BHMT mRNA levels were reduced to 29+/ 23% in established liver cirrhosis induced by bile duct ligation (BDL) as compared to controls. Enzyme assays in crude liver homogenates showed a similar reduction in BHMT activity in bile duct ligated rat livers. By Western blotting, BHMT could be detected in crude liver homogenates of control animals, but was reduced to below the limit of detection in cirrhotic livers. In conclusion, these findings establish a reduced BHMT enzyme activity in cirrhotic rat livers, which may explain the elevated plasma homocysteine levels in cirrhosis. PMID- 12757932 TI - p80 coilin, a coiled body-specific protein, interacts with ataxin-1, the SCA1 gene product. AB - Spinocerebellar ataxia type 1 (SCA1) is an autosomal-dominant neurodegenerative disorder characterized by ataxia and progressive motor deterioration. SCA1 is associated with an elongated polyglutamine tract in ataxin-1, the SCA1 gene product. Using the yeast two-hybrid system and co-immunoprecipitation experiments, we have found that p80 coilin, coiled body-specific protein, binds to ataxin-1. In further experiments with deletion mutants, we found that the C terminal regions of ataxin-1 and p80 coilin were essential for this interaction. In HeLa cells that have been co-transfected with ataxin-1 and p80 coilin, the p80 coilin protein co-localizes with ataxin-1 aggregates in the nucleoplasm. However, immunohistochemical analysis and immunofluorescence assays showed that mutant ataxin-1 aggregates do not redistribute p80 coilin's dot-like structures in the Purkinje cells of SCA1 transgenic mice. This feature of the interaction between ataxin-1 and p80 coilin suggests that p80 coilin might be implicated in altering the function of ataxin-1. PMID- 12757933 TI - Functional analysis of PCCB mutations causing propionic acidemia based on expression studies in deficient human skin fibroblasts. AB - Propionic acidemia (PA) is a recessive disorder caused by a deficiency of propionyl-CoA carboxylase (PCC), a dodecameric enzyme composed of two different proteins alpha-PCC and beta-PCC, nuclear encoded by the PCCA and PCCB genes, respectively. Mutations in either gene cause PA and to date, up to 47 different allelic variations in the PCCB gene have been identified in different populations. In this work, we describe the expression studies of 18 PCCB sequence changes in order to elucidate their functional consequences. We have used a PCCB deficient transformed fibroblast cell line to target the wild-type and mutant proteins to their physiological situation, analysing the effect of the mutations on PCC activity and protein stability. Of the 18 mutant proteins tested for activity, those carrying the L17M and A497V substitutions showed an activity similar to the wild-type one, which proves that these changes do not have any effect on protein activity. The other 16 mutant proteins exhibited two different functional behaviours, 3 retained substantial activity (K218R, R410W and N536D), and the remaining 13 proteins showed null or very low activity. Western blot analysis demonstrated instability only for the L519P, R512C and G112D mutant proteins. We have proved the pathogenicity of R67S, R165Q and G112D mutation in PCCB gene, expressed for the first time in this work. The information derived from the expression analysis is discussed in the phenotype and genotype context in order to improve the knowledge of this complex disease. PMID- 12757934 TI - Kinetic properties of the glucose 6-phosphatase of the liver from arthritic rats. AB - According to previous reports, adjuvant-induced arthritic rats present reduced activities of the hepatic glucose 6-phosphatase. A kinetic study was done in order to characterize this phenomenon. Microsomes were isolated from livers of arthritic and control rats (Holtzman strain) and the glucose 6-phosphatase was measured at various temperatures (13-37 degrees C) and glucose 6-phosphate concentrations. Irrespective of the temperature, the enzyme from arthritic rats presented a reduction of both V(max) and K(M). Detergent treatment of liver microsomes from control rats increased the activity, but no increase was found when microsomes from arthritic rats were treated in the same way. The mannose 6 phosphatase activity of detergent-treated microsomes from arthritic rats was only 25% of the activity found with detergent-treated microsomes from control rats. Without detergent treatment, the mannose 6-phosphatase activities of both control and arthritic rats were minimal. The activation energy, derived from V(max), was not changed by arthritis. In vivo arthritic rats presented higher hepatic glucose 6-phosphate concentrations, a phenomenon that is consistent with a reduced activity of glucose 6-phosphatase. It was concluded that in arthritic rats, the hydrolase is probably reduced, without a similar change in the translocase activity. PMID- 12757936 TI - Hypoxia alters expression and function of syncytin and its receptor during trophoblast cell fusion of human placental BeWo cells: implications for impaired trophoblast syncytialisation in pre-eclampsia. AB - The fundamental process of placental trophoblast cell fusion (syncytiotrophoblast formation or syncytialisation) which is a characteristic of this tissue is poorly understood. Pre-eclampsia is associated with placental hypoxia and suppressed syncytiotrophoblast formation. We therefore have studied the effect of low-oxygen tensions on the rate of cell fusion, relative abundance of mRNAs encoding syncytin and its receptor, amino acid transport system B(0), which are thought to be involved in trophoblast cell fusion (as well as the activity of amino acid transport through this system) in a cell model of syncytialisation (BeWo cells following forskolin treatment). Forskolin-induced cell fusion (determined by a quantitative flow cytometry assay) was reversibly suppressed in 2% oxygen compared to 20% oxygen. This was associated with suppressed secretion of human chorionic gonadotropin. Forskolin stimulated relatively less syncytin mRNA (determined by reverse transcription-polymerase chain reaction) in 2% than in 20% oxygen and there was no stimulation after 48 h in 2% oxygen. There was a spontaneous, time-dependent increase of amino acid transporter B(0) mRNA in vehicle, which was suppressed by 2% oxygen and by forskolin treatment in 20% oxygen. Forskolin-induced changes in amino acid transport system B(0) function were not seen in cells cultured for 48 h in 2% oxygen. These observations suggest that under conditions of low ambient oxygen, dysregulation of expression of syncytin and of its receptor may suppress the normal process of cell fusion necessary for syncytiotrophoblast formation and contributes to syncytiotrophoblast abnormalities characteristic of pre-eclampsia. PMID- 12757935 TI - Extracellular matrix and nuclear abnormalities in skeletal muscle of a patient with Walker-Warburg syndrome caused by POMT1 mutation. AB - Walker-Warburg syndrome (WWS) is an autosomal recessive disorder characterized by congenital muscular dystrophy, structural eye abnormalities and severe brain malformations. We performed an immunohistochemical and electron microscopy study of a muscle biopsy from a patient affected by WWS carrying a homozygous frameshift mutation in O-mannosyltransferase 1 gene (POMT1). alpha-Dystroglycan glycosylated epitope was not detected in muscle fibers and intramuscular peripheral nerves. Laminin alpha2 chain and perlecan were reduced in muscle fibers and well preserved in intramuscular peripheral nerves. The basal lamina in several muscle fibers showed discontinuities and detachment from the plasmalemma. Most nuclei, including myonuclei and satellite cell nuclei, showed detachment or complete absence of peripheral heterochromatin from the nuclear envelope. Apoptotic changes were detected in 3% of muscle fibers. The particular combination of basal lamina and nuclear changes may suggest that a complex pathogenetic mechanism, affecting several subcellular compartments, underlies the degenerative process in WWS muscle. PMID- 12757937 TI - Vitronectin in human breast carcinomas. AB - We have analysed the occurrence of the extracellular glycoprotein vitronectin in carcinomas and normal tissue of human breast. Immunohistochemical analysis of carcinomas revealed a strong vitronectin accumulation in extracellular matrix (ECM) around some cancer cell clusters and in the subendothelial area of some blood vessels. In normal tissue, vitronectin had a homogeneous periductal occurrence, with local accumulation much lower than that in the carcinomas. Using a new solid phase radioligand assay, the vitronectin concentrations of extracts of carcinomas and normal breast tissue were determined and found to be indistinguishable. Comparison of the vitronectin and the hemoglobin concentrations of the extracts showed that their vitronectin content was not derived from blood contamination. Vitronectin mRNA was undetectable in both carcinomas and normal tissue. We conclude that vitronectin is not synthesised locally in breast tissue but derived by leakage from vessels, followed by extracellular accumulation in patterns distinctly different in carcinomas and normal tissue. The observation of a high vitronectin content in the carcinomas and its localisation in the tissue contributes to the clarification of the role of vitronectin in tumour biology in interaction with the plasminogen activation system and integrins. PMID- 12757938 TI - Cloning genomic INGAP: a Reg-related family member with distinct transcriptional regulation sites. AB - The protein product of hamster islet neogenesis-associated protein (INGAP) cDNA induces new pancreatic islet development. Manipulation of this process provides a new therapeutic strategy for the treatment of diabetes. As regulators of INGAP gene expression are unknown over 6 kb of hamster genomic INGAP has been cloned. Sequence analysis has identified a 3 kb 5-prime region with core promoter elements that is rich in transcription factor binding sites and six exons for the coding region. Analysis of promoter activity reveals stimulus-responsive DNA elements which have been identified though deletion analysis. Comparison of transcription factor binding sites in INGAP to the related gene RegIIIdelta exposes potential sites for differential gene regulation. PMID- 12757939 TI - The phosphotriesterase gene opdA in Agrobacterium radiobacter P230 is transposable. AB - We report a transposase gene (tnpA) upstream of the opdA phosphotriesterase gene of Agrobacterium radiobacter P230, as well as inverted repeats indicative of insertion sequences, flanking the two genes. Both the tnpA gene and the inverted repeats resemble the Tn610 transposon from Mycobacterium fortuitum. Two additional putative open reading frames separate opdA and tnpA with inferred translation products with similarity to two proteins encoded on the Geobacillus stearothermophilus IS5376 transposon. To test the proposition that these genes were contained on a transposon, an artificial composite transposon was constructed. This artificial transposon was then delivered into Escherichia coli DH10beta cells. Transposition was demonstrated by the presence of opdA on the E. coli chromosome and confirmation of insertion by inverse polymerase chain reaction. The data presented suggest a possible role of transposition in the distribution of the opd/opdA genes across a wide range of soil bacteria. PMID- 12757940 TI - Rapid identification of the species of the Bacteroides fragilis group by multiplex PCR assays using group- and species-specific primers. AB - We report a rapid and reliable two-step multiplex polymerase chain reaction (PCR) assay to identify the 10 Bacteroides fragilis group species - Bacteroides caccae, B. distasonis, B. eggerthii, B. fragilis, B. merdae, B. ovatus, B. stercoris, B. thetaiotaomicron, B. uniformis and B. vulgatus. These 10 species were first divided into three subgroups by multiplex PCR-G, followed by three multiplex PCR assays with three species-specific primer mixtures for identification to the species level. The primers were designed from nucleotide sequences of the 16S rRNA, the 16S-23S rRNA intergenic spacer region and part of the 23S rRNA gene. The established two-step multiplex PCR identification scheme was applied to the identification of 155 clinical isolates of the B. fragilis group that were previously identified to the species level by phenotypic tests. The new scheme was more accurate than phenotypic identification, which was accurate only 84.5% of the time. The multiplex PCR scheme established in this study is a simple, rapid and reliable method for the identification of the B. fragilis group species. This will permit more accurate assessment of the role of various B. fragilis group members in infections and of the degree of antimicrobial resistance in each of the group members. PMID- 12757941 TI - HutC/FarR-like bacterial transcription factors of the GntR family contain a small molecule-binding domain of the chorismate lyase fold. AB - Numerous bacterial transcription factors contain a DNA-binding helix-turn-helix domain and a signaling domain, linked together in a single polypeptide. Typically, this signaling domain is a small-molecule-binding domain that undergoes a conformational change upon recognizing a specific ligand. The HutC/FarR-like transcription factors of the GntR family are one of the largest groups of transcription factors in the proteomes of most free-living bacteria. Using sensitive sequence profile analysis we show that the HutC/FarR-like transcription factors contain a conserved ligand-binding domain, which possesses the same fold as chorismate lyase (Escherichia coli UbiC gene product). This relationship suggests that the C-terminal domain of the HutC/FarR-like transcription factors binds small molecules in a cleft similar to the substrate binding site of the chorismate lyases. The sequence diversity within the predicted binding cleft of the HutC/FarR ligand-binding domains is consistent with the ability of these transcription factors to respond to diverse small molecules, such as histidine (HutC), fatty acids (FarR), sugars (TreR) and alkylphosphonate (PhnF). UbiC-like chorismate lyases function in the ubiquinone biosynthesis pathway, and have characteristic charged, catalytic residues. Genome comparisons reveal that chorismate lyase orthologs are found in several bacteria, chloroplasts of eukaryotic algae and euryarchaea. In contrast, the GntR transcription regulators lack the conserved catalytic residues of the chorismate lyases, and have so far been detected only in bacteria. An ancestral, generic small-molecule-binding domain appears to have given rise to the enzymatic and non catalytic ligand-binding versions of the same fold under the influence of different selective pressures. PMID- 12757942 TI - Null mutations in the essential gene yrfF (mucM) are not lethal in rcsB, yojN or rcsC strains of Salmonella enterica serovar Typhimurium. AB - Insertion of factor MudJ in the intergenic region between divergent genes yrfF and yrfE, at centisome 76 in the genome of Salmonella enterica serovar Typhimurium LT2, confers the characteristics recently described for mucM mutants, i.e. mucoidy and resistance to mecillinam. Cloning of the intergenic region plus either the yrfF or the yrfE gene in a multicopy plasmid showed that only the plasmid carrying the yrfF gene complemented mucM mutants, thus suggesting that mucM mutations are in fact yrfF mutations. A null yrfF mutation obtained by insertion of a kanamycin cassette into the yrfF open reading frame (yrfF28::Kan) produced abortive colonies when transduced to a wild-type strain but was normally accepted by rcsB, rcsC or yojN strains. Neither mutations preventing synthesis of the capsular exopolysaccharide colanic acid (cps, galE) nor rcsA mutations, which reduce expression of cps genes, conferred tolerance to the lethal yrfF28::Kan mutation. Spontaneous suppressor mutations arose very frequently in abortive yrfF28::Kan colonies, and all of them affected either rcsC, yojN, or rcsB genes. Thus, the lethal effect caused by inactivation of gene yrfF appears to be mediated by a function that is dependent on the rcsC-yojN-rcsB phosphorelay system but does not involve synthesis of colanic acid. PMID- 12757943 TI - Sulerythrin, the smallest member of the rubrerythrin family, from a strictly aerobic and thermoacidophilic archaeon, Sulfolobus tokodaii strain 7. AB - A protein corresponding to the N-terminal domain of rubrerythrin was isolated from a strictly aerobic archaeon, Sulfolobus tokodaii strain 7. The molecular mass was found to be 15.8 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, 16278 Da by time-of-flight mass spectrometry and 34.5 kDa by gel filtration chromatography, suggesting that the protein is dimeric. Two mol iron and 1-2 mol zinc mol(-1) protein were detected. On addition of the azide ion, the absorption spectrum was greatly affected. The far UV circular dichroism spectrum suggested that the protein was mostly composed of alpha-helices. The N-terminal sequence completely matched the open reading frame, st2370, recently found on genome analysis of the organism. The protein was homologous to rubrerythrin but lacked a C-terminal rubredoxin domain. It was found in the genus Sulfolobus and therefore named sulerythrin; it is the smallest and first aerobic member of the rubrerythrin family. PMID- 12757944 TI - Involvement of enterobactin in norepinephrine-mediated iron supply from transferrin to enterohaemorrhagic Escherichia coli. AB - Exposure of bacteria to members of the stress-associated family of catecholamine hormones, principally norepinephrine, has been demonstrated to increase both growth and production of virulence-related factors. Mutation of genes for enterobactin synthesis and uptake revealed an absolute requirement for enterobactin in norepinephrine-stimulated growth of Escherichia coli O157:H7. The autoinducer produced by norepinephrine-stimulated E. coli could not substitute for enterobactin. We also demonstrate that norepinephrine promotes iron shuttling between transferrin molecules, thereby enabling the bacterial siderophore enterobactin to more readily acquire iron for growth. These results suggest one of the possible mechanisms by which the hormonal output of stress may affect enterohaemorrhagic E. coli pathogenicity. PMID- 12757945 TI - Purification and characterization of an endochitinase produced by Colletotrichum gloeosporioides. AB - The phytopathogenic fungus Colletotrichum gloeosporioides was analyzed for chitinase activity, the best production occurring at the fourth day. A 43 kDa endochitinase with specific activity of 413 U microg(-1) protein was purified corresponding to a 75% yield. The optima of temperature and pH for the enzyme were 50 degrees C and pH 7.0, respectively. The enzyme showed a high stability at 50 degrees C and pH 7.0. Values of pH from 5.0 up to 7.0 gave, at least, 50% of maximum activity, suggesting a biotechnological application. Further studies are in progress to determine the possible use of this endochitinase in biological control. PMID- 12757946 TI - Differentiation of Borrelia burgdorferi sensu lato through groEL gene analysis. AB - The nucleotide sequences (310 bp) of the groEL gene, which encode the 60-kDa heat shock protein GroEL from 31 reference strains of Borrelia were determined and compared. More than 92.3% similarity was observed among Borrelia burgdorferi sensu lato strains. In the phylogenetic tree constructed with the maximum likelihood method, each species of B. burgdorferi sensu lato was differentiated as a distinct entity. We developed polymerase chain reaction-restriction fragment length polymorphism analysis using a specific single amino acid variation [N(213) (AAT)-->S (AGC or AGT)] between B. burgdorferi sensu stricto strains and the other B. burgdorferi sensu lato strains. These results showed that the groEL gene is useful for differentiation of B. burgdorferi sensu lato. PMID- 12757947 TI - Origin and evolution of transmembrane Chl-binding proteins: hydrophobic cluster analysis suggests a common one-helix ancestor for prokaryotic (Pcb) and eukaryotic (LHC) antenna protein superfamilies. AB - All chlorophyll (Chl)-binding proteins constituting the photosynthetic apparatus of both prokaryotes and eukaryotes possess hydrophobic domains, corresponding to membrane-spanning alpha-helices (MSHs). Hydrophobic cluster analysis of representative members of the different Chl protein superfamilies revealed that all Chl proteins except the five-helix reaction center II proteins and the small subunits of photosystem I possess related domains. As a major conclusion, we found that the eukaryotic antennae likely share a common precursor with the prokaryotic Chl a/b antennae from Chl-b-containing oxyphotobacteria. From these data, we propose a global scheme for the evolution of these proteins from a one MSH ancestor. PMID- 12757948 TI - CNE, a collagen-binding protein of Streptococcus equi. AB - Streptococcus equi subspecies equi is an important horse pathogenic bacterium causing a serious disease called strangles. Using bioinformatics we identified a gene denoted cne (gene encoding collagen-binding protein from S. equi) coding for a novel potential virulence factor of this species called protein CNE. The protein is composed of 657 amino acids and has the typical features found in cell surface-anchored proteins in Gram-positive bacteria. CNE displays amino acid sequence similarities to the previously well-studied collagen-binding protein CNA from Staphylococcus aureus, a proven virulence factor in septic arthritis. Based on similarity to CNA the structure of the mature CNE protein can be divided into an N-terminal A domain and a C-terminal B domain. The highest similarity between CNA and CNE is found in the A domains. The A domain in CNA is known to be the collagen-binding domain. Two parts of cne were amplified using polymerase chain reaction (PCR) and ligated into an expression vector, and recombinant CNE proteins were produced in Escherichia coli. The purified CNE proteins were shown to display collagen-binding activity in a Western ligand blot and to inhibit collagen binding to cells of subsp. equi and to CNE-coated microtitre wells. Furthermore, the A domain of CNE was sufficient for binding collagen, and was shown to compete for the same site on collagen as CNA in inhibition studies. Using PCR, the cne gene was detected in all studied strains of subsp. equi and S. equi subsp. zooepidemicus. PMID- 12757949 TI - Immunochemical detection of Salmonella group B, D and E using an optical surface plasmon resonance biosensor. AB - A surface plasmon resonance biosensor (Biacore) was used to detect Salmonella through antibodies reacting with Salmonella group A, B, D and E (Kauffmann-White typing). In the assay designed, anti-Salmonella antibodies immobilized to the biosensor surface were allowed to bind injected bacteria followed by a pulse with soluble anti-Salmonella immunoglobulins to intensify the signal. No significant interference was found for (mixtures of) 30 non-Salmonella serovars at 10(9) CFU ml(-1). A total of 53 Salmonella serovars were successfully detected at 1 x 10(7) CFU ml(-1), except those of groups C, G, L and P, as expected. The cut-off point was determined with an equicellular mixture of Salmonella enteritidis and Salmonella typhimurium at a final amount of 1.7 x 10(3) CFU per test portion. Although further work is needed to cover the detection of all relevant Salmonella serovars in food-producing animals and food products, this work demonstrates the merits of this alternative biosensor approach in terms of automation, sensitivity, specificity, simple handling and limited hands-on time. PMID- 12757950 TI - Expression of putative pathogenicity-related genes in Xylella fastidiosa grown at low and high cell density conditions in vitro. AB - Xylella fastidiosa is the causal agent of economically important plant diseases, including citrus variegated chlorosis and Pierce's disease. Hitherto, there has been no information on the molecular mechanisms controlling X. fastidiosa-plant interactions. To determine whether predicted open reading frames (ORFs) encoding putative pathogenicity-related factors were expressed by X. fastidiosa 9a5c cells grown at low (LCD) and high cell density (HCD) conditions in liquid modified PW medium, reverse Northern blot hybridization and reverse transcription-polymerase chain reaction (RT-PCR) experiments were performed. Our results indicated that ORFs XF2344, XF2369, XF1851 and XF0125, encoding putative Fur, GumC, a serine protease and RsmA, respectively, were significantly suppressed at HCD conditions. In contrast, ORF XF1115, encoding putative RpfF, was significantly induced at HCD conditions. Expressions of ORFs XF2367, XF2362 and XF0290, encoding putative GumD, GumJ and RpfA, respectively, were detected only at HCD conditions, whereas expression of ORF XF0287, encoding putative RpfB was detected only at LCD conditions. Bioassays with an Agrobacterium traG::lacZ reporter system indicated that X. fastidiosa does not synthesize N-acyl-homoserine lactones, whereas bioassays with a diffusible signal factor (DSF)-responsive Xanthomonas campestris pv. campestris mutant indicate that X. fastidiosa synthesizes a molecule similar to DSF in modified PW medium. Our data also suggest that the synthesis of the DSF like molecule and fastidian gum by X. fastidiosa is affected by cell density in vitro. PMID- 12757951 TI - Sequence analysis and biochemical characterisation of chromosomal CAV-1 (Aeromonas caviae), the parental cephalosporinase of plasmid-mediated AmpC 'FOX' cluster. AB - Aeromonas caviae CIP 74.32 was resistant to amoxicillin, ticarcillin and cephalothin, and susceptible to cefoxitin, cefotaxime, ceftazidime, aztreonam and imipenem. This strain produced a cephalosporinase (pI 7.2) and an oxacillinase (pI 8.5). The cephalosporinase gene cav-1 was cloned and sequenced. Unlike A. caviae donor, Escherichia coli pNCE50 transformant producing CAV-1 beta-lactamase was resistant to cefoxitin. The deduced protein sequence CAV-1 contained 382 amino acids, and shared >96% homology with FOX-1 to FOX-5 cephalosporinase. CAV-1 presented only two amino acid substitutions (Thr270Ser and Arg271Ala) with FOX-1. CAV-1 is the chromosomal putative ancestor of the FOX family, a cluster of class C/group 1 plasmidic cephalosporinases spreading in Klebsiella and E. coli clinical isolates via conjugative plasmids. PMID- 12757953 TI - Isolation and mutation site determination of the temperature-sensitive murB mutants of Staphylococcus aureus. AB - The murB gene encodes UDP-N-acetylenolpyruvylglucosamine reductase and functions in bacterial peptidoglycan biosynthesis. A plasmid carrying the murB gene restored the temperature-sensitive growth of six Staphylococcus aureus mutants, in which peptidoglycan biosynthesis stopped at a restrictive temperature. Specific activity of UDP-N-acetylenolpyruvylglucosamine reductase in extracts from the mutants was lower than that from wild-type cells. Nucleotide sequence determination revealed that each mutant had a single amino acid substitution in the murB gene and five of six mutations were located within domain 3, where the proposed substrate binding site is located. These results suggest that the murB gene is essential for growth of S. aureus and that domain 3 is important for the MurB activity. PMID- 12757952 TI - A model of meningococcal bacteremia after respiratory superinfection in influenza A virus-infected mice. AB - We developed a model of sequential influenza A virus (IAV)-Neisseria meningitidis serogroup C (Nm) infection in BALB/c mice. Mice infected intranasally with a sublethal IAV dose (260 pfu) were superinfected intranasally with Nm. Fatal meningococcal pneumonia and bacteremia were observed in IAV-infected mice superinfected with Nm on day 7, but not in those superinfected on day 10. The susceptibility of mice to Nm superinfection was correlated with the peak interferon-gamma production in the lungs and decrease in IAV load. After Nm challenge, both IAV-infected and uninfected control mice produced the inflammatory cytokines interleukin (IL)-1 and IL-6. However, IL-10 was detected in susceptible mice superinfected on day 7 after IAV infection, but not in resistant mice. This model of dual IAV-Nm infection was also used to evaluate the role of bacterial virulence factors in the synthesis of the capsule. A capsule defective mutant was cleared from the lungs, whereas a mutant inactivated for the crgA gene, negatively regulating expression of the pili and capsule, upon contact with host cells, retained invasiveness. Therefore, this model of meningococcal disease in adult mice reproduces the pathogenesis of human meningococcemia with fatal sepsis, and is useful for analyzing known or new genes identified in genomic studies. PMID- 12757955 TI - Optimization of the green fluorescent protein (GFP) expression from a lactose inducible promoter in Lactobacillus casei. AB - An expression vector for Lactobacillus casei has been constructed containing the inducible lac promoter and the gene encoding ultraviolet visible green fluorescent protein (GFP(UV)) as reporter. Different conditions to grow L. casei were assayed and fluorescence as well as total protein synthesized were quantified. The maintenance of neutral pH had the greatest incidence on GFP(UV) expression, followed by aeration and a temperature of 30 degrees C. Environmental factors favoring GFP(UV) accumulation did not exactly correlate with those enhancing fluorescence. Therefore, oxygenation, by stirring the culture, had the greatest influence on the proportion of fluorescent protein, which is in accordance with the structural requirements of this protein. The highest yield obtained was 1.3 microg of GFP per mg of total protein, from which 55% was fluorescent. PMID- 12757954 TI - 60Co irradiation of Shiga toxin (Stx)-producing Escherichia coli induces Stx phage. AB - Shiga toxin (Stx)-producing Escherichia coli (STEC), an important cause of hemolytic uremic syndrome, was completely killed by (60)Co irradiation at 1 x l0(3) gray (1 kGy) or higher. However, a low dose of irradiation (0.1-0.3 kGy) markedly induced Stx phage from STEC. Stx production was observed in parallel to the phage induction. Inactivation of Stx phage required a higher irradiation dose than that for bacterial killing. Regarding Stx, cytotoxicity was susceptible to irradiation, but cytokine induction activity was more resistant than Stx phage. The findings suggest that (1). although (60)Co irradiation is an effective means to kill the bacteria, it does induce Stx phage at a lower irradiation dose, with a risk of Stx phage transfer and emergence of new Stx-producing strains, and (2). irradiation differentially inactivates some activities of Stx. PMID- 12757956 TI - Multiplex PCR using 16S rRNA gene-targeted primers for the identification of bifidobacteria from human origin. AB - Three multiplex polymerase chain reactions (PCRs) targeted on Bifidobacterium and related species were designed to identify human species. The selected primers yielded amplified products of various sizes, each specific for a species. Three to four pairs were gathered in one PCR reaction and their specificity under multiplex conditions was confirmed using DNA from 26 reference strains. Using this technique on unidentified faecal strains, B. bifidum, B. longum and B. breve species were commonly recovered in infants while B. adolescentis, B. catenulatum/B. pseudocatenulatum continuum and B. longum species were predominant in adults. Thus, a single PCR can provide the assignment of a strain to one these species, reducing the number of PCR reactions and hands-on time for the identification of human isolates of bifidobacteria. Moreover, this technique is also applicable for the in situ detection of bifidobacteria in DNA extracts from human stools. PMID- 12757957 TI - The VirR/VirS regulatory cascade affects transcription of plasmid-encoded putative virulence genes in Clostridium perfringens strain 13. AB - We analyzed the transcriptional regulation of the putative virulence genes encoded on the plasmid pCP13 from Clostridium perfringens strain 13. The transcription of the beta2-toxin (cpb2) and possible collagen adhesin (cna) genes were regulated in both a positive and negative manner, respectively, by the two component VirR/VirS system. The secondary regulator of the VirR/VirS system, VR RNA, also affects the expression of both of these genes in the same fashion as the VirR/VirS system. This indicates that the global regulatory cascade of the VirR/VirS system controls the expression of virulence genes located on the plasmid, as well as those found chromosomally in C. perfringens strain 13. PMID- 12757958 TI - Species level identification of conifer associated Ceratocystis sapstain fungi by PCR-RFLP on a beta-tubulin gene fragment. AB - The genus Ceratocystis includes several morphologically similar species commonly found as agents of sapstain in coniferous trees. In this paper we describe a simple and reliable polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique that aids in the identification and differentiation of these fungi. PCR was used to amplify a 1.3-kb fragment of the beta-tubulin gene from C. coerulescens, C. pinicola, C. douglassi, C. resinifera, C. rufipenni, C. polonica and C. adiposa. The PCR amplicon from representative isolates was sequenced. This information was utilized to select restriction enzymes that generated species-specific RFLP patterns. This approach was tested on our collection of over 200 Ceratocystis isolates and identified the fungi with a high level of confidence, reducing the time needed to identify these species by classical methods. PMID- 12757959 TI - Production of landomycins in Streptomyces globisporus 1912 and S cyanogenus S136 is regulated by genes encoding putative transcriptional activators. AB - The regulatory genes lanI and lndI have been cloned from the landomycin A (LaA) producer Streptomyces cyanogenus S136 and from the landomycin E (LaE) producer Streptomyces globisporus 1912, respectively and both have been sequenced. A culture of S. globisporus I2-1 carrying a disrupted lndI gene did not produce LaE and other related intermediates. Complementation of S. globisporus I2-1 with either the lndI or lanI gene reconstituted LaE production indicating that LanI and LndI are involved in activation of structural genes in the respective clusters. Structural features of these regulatory genes are discussed. PMID- 12757960 TI - Treatment of adolescent tobacco smokers: issues and opportunities for exposure reduction approaches. AB - The cycle of tobacco dependence typically begins with the initiation of tobacco use during adolescence. Many teenagers try to quit smoking, fail and subsequently desire treatment for their tobacco dependence. Adolescents do not currently benefit from the same level of societal support for quit attempts as adults, and they may be less motivated for total cessation despite the short and long-term health consequences of smoking. Overall, the combination of low participation, high attrition and low complete cessation rates for adolescent smokers in treatment prompts the consideration of alternative treatment endpoints. It is likely that interactions among the processes of child and adolescent development, smoke exposure and trajectory influence patterns of tobacco use and treatment for tobacco dependence in adolescents. A rational framework is needed to integrate the study of these dynamic interactions to address tobacco dependence among youth from an exposure reduction, in addition to a cessation, perspective. This paper considers the issues and potential implications of tobacco exposure reduction therapy as an intermediate treatment goal for adolescent smokers who are dependent or dependence-prone, but for whom initial treatment interventions do not yield complete tobacco cessation. PMID- 12757961 TI - Behavioral and subjective effects of marijuana following partial sleep deprivation. AB - This study tested whether performance would be more impaired when marijuana use followed partial sleep deprivation (PSD) than when marijuana use followed a typical night of sleep. Seven recreational marijuana users (mean 15 of last 30 days) completed six test sessions in a double-blind randomized within-subject design. Each session began with an overnight stay in a sleep laboratory. Bed and wake times were calculated from mean data on individual sleep diaries. Time-in bed was either regular (mean=8.2 h) or shortened (first 65% of regular time-in bed deprived). At 3 and 5 h after waking, daytime sleepiness was measured with self-report questionnaires and a sleep latency test. Approximately 6.5 h after waking, subjects smoked a marijuana cigarette (0.003, 2, or 3.5% delta-9 tetrahydrocannabinol [THC]). Test batteries were completed 2, 62, and 122 min after smoking ended. Sleepiness was significantly greater following PSD than after regular sleep. Following regular sleep, heart rate increases with active THC doses were comparable, but heart rate with 2% THC was significantly less elevated following PSD. Ratings of "impaired" and "stoned" increased with both THC doses after regular sleep and were further increased with 3.5% THC after PSD. High-potency marijuana increased body sway similarly across sleep conditions. There were no significant effects of marijuana or PSD, alone or in combination, on brake latency. Thus, while PSD increased the dose-dependence of THC effects on heart rate and subjective impairment, it did not enhance the effects of marijuana on standing balance and brake latency. PMID- 12757962 TI - Recanting of substance use reports in a longitudinal prevention study. AB - We analyzed recanting of substance use reports for lifetime use of alcohol, alcohol to get drunk, cigarettes, marijuana and cocaine in an 8-wave panel study designed to evaluate the Drug Abuse Resistance Education program in the State of Illinois. Although this phenomenon has been identified elsewhere, the current analysis of recanting is a unique attempt to track this behavior over the entire course of adolescence. Overall, rates of recanting for specific drugs were extremely high, ranging from 45% for lifetime reports of alcohol use to 81% for lifetime reports of cocaine. Most recanting occurred in the wave immediately following the wave of first disclosure. Paralleling results from other studies, race/ethnicity was an important correlate of recanting in both bivariate and multivariate analyses. African American respondents had higher rates of recanting than White subjects. Even after controlling for the number of follow-up waves, the later the wave of first disclosed lifetime drug use, the lower the probability that drug use would be recanted ever (for all substances) or in the wave immediately following first disclosure (for reports of ever having been drunk or for lifetime marijuana or cocaine use). Alternative causes for this phenomenon are discussed. Implications for the design and interpretation of multiwave school-based panel surveys targeted toward adolescents are also addressed. PMID- 12757963 TI - Discriminative stimulus effects of (-)-ephedrine in rats: analysis with catecholamine transporter and receptor ligands. AB - A drug discrimination procedure was used to examine the neuropharmacology of (-) ephedrine (5 mg/kg), a sympathomimetic amine found in a variety of dietary supplements. (-)-Ephedrine has caused concern because of its use as a precursor in the manufacture of street drugs (e.g. methamphetamine) and its potential for abuse and toxicity. In the present study, the catecholamine reuptake inhibitors mazindol and nomifensine, the norepinephrine (NE) reuptake inhibitor desipramine, and the dopamine D(2)-like (e.g. D(2), D(3) and D(4)) agonist quinpirole substituted for (-)-ephedrine (>/=80% (-)-ephedrine-lever responding). The NE reuptake inhibitor nisoxetine, the D(1)-like (e.g. D(1) and D(5)) agonists (+/-) SKF 38393 and SKF 82958, and the mixed D(1)-/D(2)-like agonist apomorphine occasioned intermediate levels of responding (50-79% (-)-ephedrine-lever responding). The (-)-ephedrine cue was antagonized by the D(1)-like antagonist SCH 23390 and the alpha(1)-adrenoceptor antagonist prazosin as well as the D(2) like antagonists (-)-eticlopride and haloperidol, although only at doses that disrupted responding in some rats. The discriminative stimulus effects of a small dose of (-)-ephedrine (1.25 mg/kg) were enhanced by the alpha(2)-adrenoceptor antagonist idazoxan and to a lesser extent by the beta-adrenoceptor antagonist ( )-propranolol. However, the alpha(2)-adrenoceptor agonist clonidine (0.04 mg/kg) did not attenuate the (-)-ephedrine stimulus. These results suggest that D(1)-, D(2)-like, and alpha(1)-adrenergic receptors mediate the discriminative stimulus effects of (-)-ephedrine. Substitution of desipramine for (-)-ephedrine and not for some other stimulants suggests that NE transmission is a prominent feature of the (-)-ephedrine discriminative stimulus, and that NE underlies therapeutic and abuse-related effects of (-)-ephedrine that diverge from those of other stimulants. PMID- 12757965 TI - Changes in patterns of drug injection concurrent with a sustained reduction in the availability of heroin in Australia. AB - Between 1996 and 2000, heroin was the drug most frequently injected in Australia, and viable heroin markets existed in six of Australia's eight jurisdictions. In 2001, there was a dramatic and sustained reduction in the availability of heroin that was accompanied by a substantial increase in its price, and a 14% decline in the average purity of seizures analysed by forensic laboratories. The shortage of heroin constitutes a unique natural experiment within which to examine the impact of supply reduction. This paper reviews one important correlate of the shortage, namely changes in patterns of illicit drug injection. A number of studies have consistently suggested that between 2000 and 2001, there was a sizeable decrease in both prevalence and frequency of heroin injection among injecting drug users. These changes were accompanied by increased prevalence and frequency of stimulant injection. Cocaine was favoured in NSW, the sole jurisdiction in which a cocaine market was established prior to the heroin shortage; whereas methamphetamine predominated in other jurisdictions. Some data suggest that, at least in the short-term, some drug injectors left the market altogether subsequent to the reduced heroin availability. However, the findings that (1) some former heroin users switched their drug preference to a stimulant; and (2) subsequently attributed this change to the reduced availability of heroin, suggests that reducing the supply of one drug may serve to increase the use of others. Given the differential harms associated with the use of stimulants and opiates, this possibility has grave implications for Australia, where the intervention and treatment system is designed primarily to accommodate opiate use and dependence. PMID- 12757964 TI - A pilot trial of olanzapine for the treatment of cocaine dependence. AB - BACKGROUND: Multiple lines of evidence suggest both dopaminergic and serotonergic involvement in the reinforcing effects of cocaine. Medications such as olanzapine, which block dopamine D2 receptors, as well as serotonin receptors 5HT2A and 5HT2C may be able to reduce cocaine use in cocaine dependent patients by reducing the euphoric effects of cocaine and attenuating cocaine craving. METHODS: This was a 12-week, double blind, placebo controlled, pilot trial involving 30 cocaine dependent subjects. Subjects received either olanzapine (10 mg/day) or identical placebo. Outcome measures included treatment retention, qualitative urine benzoylecgonine tests, cocaine craving, clinical global impression scores, and results from the addiction severity index. RESULTS: Treatment retention was slightly, but significantly, better in the placebo treated subjects. Placebo-treated subjects were more likely to be abstinent from cocaine during the trial compared to olanzapine-treated subjects, based on urine benzoylecgonine results. Olanzapine was not superior to placebo in any outcome measure. CONCLUSIONS: The results of this trial do not support the usefulness of olanzapine for the treatment of cocaine dependence. In fact, olanzapine may worsen cocaine treatment outcome. PMID- 12757966 TI - Buprenorphine dosing regime for inpatient heroin withdrawal: a symptom-triggered dose titration study. AB - The study aimed to identify the range of buprenorphine doses required to comfortably alleviate symptoms in patients undergoing inpatient heroin withdrawal using a symptom-triggered titration dosing regime, and to identify the patient characteristics that impact upon the buprenorphine dose requirements. The study was conducted in two Australian inpatient withdrawal units, recruiting 63 dependent, injecting heroin users with no recent methadone treatment, dependence on other drugs, or other active medical or psychiatric conditions. In a single (patient) blinded case series, placebo or 2 mg sublingual buprenorphine tablets was administered four times a day according to severity of withdrawal (assessed with Subjective Opiate Withdrawal Scale). Up to 16 mg buprenorphine was available over the first 4 days of the admission, up to 8 mg on day 5, and placebo continued until day 6. Thirty-two subjects completed the dosing regime, with mean (+/-S.D.) daily doses of 3.8+/-2.8 on day 1, 5.8+/-3.2 on day 2, 4.8+/-3.3 on day 3, 2.3+/-2.6 on day 4, 0.8+/-1.3 on day 5, and a total dose of 17.4+/-9.7. Higher buprenorphine doses were required by those patients with more severe psychosocial dysfunction, women, those with more frequent heroin use, and those with more severe dependence on heroin at intake. A dosing regime using sublingual buprenorphine tablets for short inpatient heroin withdrawal is proposed. PMID- 12757967 TI - A genome-wide search for quantitative trait loci influencing substance dependence vulnerability in adolescence. AB - This study describes results from a genome-wide search for quantitative trait loci (QTL) influencing substance dependence vulnerability in adolescence. We utilized regression-based multipoint (and single-point) QTL mapping procedures designed for selected sibpair samples. Selected sibling pairs included 250 proband-sibling pairs from 192 families. Clinical probands (13-19 years of age) were drawn from consecutive admissions to substance abuse treatment facilities in the Denver metropolitan area; siblings of probands ranged in age from 12 to 25 years. In addition to the selected sample, a community-based sample of 3676 adolescents and young adults were utilized to define a clinically-significant, heritable, age- and sex-normed index of substance dependence vulnerability-a priori and independent of our linkage results. Siblings and their parents were genotyped for 374 STR micro-satellite markers distributed across the 22 autosomes (average inter-marker distance=9.2 cM). Non-parametric single-point linkage results indicated 17 markers on 11 chromosomes with nominally significant tests of linkage; six markers with LOD scores greater than 1.0 and one marker (D3S1614) with a LOD score of 2.2. Multipoint mapping corroborated two locations and provided preliminary evidence for linkage to regions on chromosome 3q24-25 (near markers D3S1279 and D3S1614) and chromosome 9q34 (near markers D9S1826 and D9S1838). PMID- 12757968 TI - Long-term outcome after benzodiazepine withdrawal treatment in subjects with complicated dependence. AB - BACKGROUND: The study aimed to monitor subjects with benzodiazepine (BZ) dependence after withdrawal treatment in order to evaluate long-term outcome and predictors of remaining BZ-free. Subjects with high-dose dependence or co occurring alcohol problems were not excluded. METHOD: Seventy-six participants in an earlier, randomized, controlled trial of outpatient BZ discontinuation were interviewed, and documents from their treatment settings obtained, along with urine and serum samples for BZ use. Long-term outcomes for a cognitive-behavioral treatment group and a treatment-as-usual group were measured. RESULTS: BZ discontinuation treatment outcomes were maintained in both treatment groups. No between-group differences were found. At the end of the study 25% of the subjects were BZ-free, and the median dose decrease from pre-treatment levels was 16.1 mg in diazepam equivalents. Subjects with pre-treatment doses exceeding 40 mg were able to maintain their doses at therapeutic levels through the follow-up. Pre treatment low BZ dose, no previous withdrawal attempts, and high life satisfaction predicted success in staying BZ-free. CONCLUSIONS: In subjects with complicated BZ dependence, the benefits of BZ discontinuation treatment may persist, but more studies are needed. PMID- 12757970 TI - A partner's drug-using status impacts women's drug treatment outcome. AB - The role that male sexual partners play in the treatment outcome of drug dependent pregnant women deserves greater attention. Pregnant women enrolled in a comprehensive treatment program with drug-free (n=85) or drug-using (n=82) male sexual partners completed a relationship survey and were compared on partner and psychosocial variables. Compared with male drug-free partners, male drug-using partners had more unemployment and more current legal involvement, less education, were less likely to be supportive of the pregnant woman's recovery efforts and were more likely to give them money to buy drugs. Male drug-free partners also had fewer medical, dental, legal and transportation needs than male drug-using partners. Data from treatment retention suggests that women with male drug-using partners are retained in a comprehensive treatment for a shorter time than women with male drug-free partners. A male partner's drug-using status should be considered when treating pregnant drug dependent women. PMID- 12757971 TI - Telomeres, telomerase and the cancer cell: an introduction. PMID- 12757969 TI - Desipramine and contingency management for cocaine and opiate dependence in buprenorphine maintained patients. AB - Co-dependence on opiates and cocaine occurs in about 60% of patients entering methadone treatment and has a poor prognosis. However, we recently found that desipramine (DMI) could be combined with buprenorphine to significantly reduce combined opiate and cocaine use among these dually dependent patients. Furthermore, contingency management (CM) has been quite potent in reducing cocaine abuse during methadone maintenance. To test the efficacy of combining CM with these medications we designed a 12-week, randomized, double blind, four cell trial evaluating DMI (150 mg/day) or placebo plus CM or a non-contingent voucher control in 160 cocaine abusers maintained on buprenorphine (median 16 mg daily). Cocaine-free and combined opiate and cocaine-free urines increased more rapidly over time in those treated with either DMI or CM, and those receiving both interventions had more drug-free urines (50%) than the other three treatment groups (25-29%). Self reported opiate and cocaine use and depressive and opioid withdrawal symptoms showed no differences among the groups and symptom levels did not correlate with urine toxicology results. Lower DMI plasma levels (average 125 ng/ml) were associated with greater cocaine-free urines. DMI and CM had independent and additive effects in facilitating cocaine-free urines in buprenorphine maintained patients. The antidepressant appeared to enhance responsiveness to CM reinforcement. PMID- 12757972 TI - Biochemical aspects of telomerase function. AB - Arthur Kornberg "never met a dull enzyme" (For the Love of Enzymes: The Odyssey of a Biochemist, Harvard University Press, 1989) and telomerase is no exception. Telomerase is a remarkable polymerase that uses an internal RNA template to reverse-transcribe telomere DNA, one nucleotide at a time, onto telomeric, G-rich single-stranded DNA. In the 17 years since its discovery, the characterization of telomerase enzyme components has uncovered a highly conserved family of telomerase reverse transcriptases that, together with the telomerase RNA, appear to comprise the enzymatic core of telomerase. While not as comprehensively understood as yet, some telomerase-associated proteins also serve crucial roles in telomerase function in vivo, such as telomerase ribonudeoprotein (RNP) assembly, recruitment to the telomere, and the coordination of DNA replication at the telomere. A selected overview of the biochemical properties of this unique enzyme, in vitro and in vivo, will be presented. PMID- 12757973 TI - Alternative lengthening of telomeres, telomerase, and cancer. AB - Telomere length may be maintained in cancer cells by telomerase or an alternative lengthening of telomeres (ALT) mechanism. Low levels of telomerase activity have been detected in some normal somatic cells and presumably some types of normal cells also have low levels of an ALT-like activity. It is hypothesized here that inherited abnormalities of these and other aspects of telomere maintenance may contribute to cancer and ageing. The telomere length maintenance mechanisms are similar in that activation of each is associated with immortalization. They may also confer other properties on cancer cells, however, and the nature of these additional properties may be different for telomerase and ALT. It is expected that these similarities and differences will have implications for prognosis and treatment. PMID- 12757974 TI - Telomerase and tumorigenesis. AB - The unique biology of telomeres and telomerase plays important roles in many aspects of mammalian cell physiology. Over the past decade, several lines of evidence have confirmed that the maintenance of telomeres and telomerase participate actively in the pathogenesis of human cancer. Specifically, activation of telomerase is strongly associated with cancer, and recent observations confirm that telomeres and telomerase perform important roles in both suppressing and facilitating malignant transformation by regulating genomic stability and cell lifespan. In addition, recent evidence suggests that telomerase activation contributes to tumorigenesis independently of its role in maintaining telomere length. Here we review recent developments in our understanding of the relationships among telomeres, telomerase, and cancer. PMID- 12757975 TI - Telomere-driven genomic instability in cancer cells. AB - Telomeres, the ends of linear chromosomes, play a major role in the maintenance of genome integrity. Telomerase or alternative lengthening of telomeres (ALT) mechanisms exist in most cancer cells in order to stabilize telomere length by the addition of telomeric repeats. Telomere loss can be dramatically mutagenic. Chromosomes lacking one telomere remain unstable until they are capped, generating chromosomal instability, gene amplification via breakage/fusion/bridge (B/F/B) cycles and resulting in chromosome imbalances. The chronology of the occurrence of gene amplification and chromosome imbalances detected in human tumors is still unknown. All of the aberrations that occur prior to, during or after activation of a telomere maintenance mechanism promote the development of cancer. PMID- 12757976 TI - Telomeres in cancer and aging: lessons from the mouse. AB - The analysis of mice deficient in telomerase activity has allowed to directly test the relevance of telomeres and telomerase in tumor development and aging in the context of a mammalian organism. More recently, mice with impaired telomere capping due to abrogation of telomere-binding proteins have been also characterized. Here, I will discuss these studies, as well as their implications for putative therapies based of telomerase inhibition of telomerase re introduction for cancer or age-related diseases, respectively. PMID- 12757977 TI - Telomere repeat binding factors: keeping the ends in check. AB - Per definition, a linear chromosome contains two ends, two sites, which by analogy to double-stranded breaks, might be expected to induce cell cycle checkpoints. The fact that cells divide without inducing such checkpoints suggests that telomeres, the natural ends of linear chromosomes, have the ability to suppress checkpoint activation. This suppression takes place at a number of levels. The TTAGGG repeats of human telomeric DNA recruit telomere specific proteins, among them the telomere repeat binding factors TRF1 and TRF2. These proteins, along with their interaction partners, reorganize the linear chromosome end into a t loop, a protected structure, which hides the very end of the chromosome. Here it is discussed how mammalian telomeres differ from DNA breaks, and what methods they use to prevent checkpoint activation. PMID- 12757978 TI - Human epithelial cell immortalization as a step in carcinogenesis. AB - Human epithelial cells encounter two senescence barriers that enforce a limited proliferative potential. A first barrier is mediated by the retinoblastoma protein, and can be overcome by multiple types of errors, many of which are observed in human cancers. A second, extremely stringent telomere-dependent barrier, is a consequence of repression of telomerase activity. Although relieved by ectopic hTERT expression, the nature of the errors required to overcome this latter barrier during in vivo carcinogenesis have not yet been defined. Attainment of immortality and telomerase reactivation are crucial to human carcinoma development; the derangements responsible for attainment of immortality may be rate-limiting and permissive for further progression to malignancy. PMID- 12757979 TI - Telomerase inhibition as cancer therapy. AB - A number of different approaches have been developed to inhibit telomerase activity in human cancer cells. Different components and types of inhibitors targeting various regulatory levels have been regarded as useful for telomerase inhibition. Most methods, however, rely on successive telomere shortening. This process is very slow and causes a long time lag between the onset of inhibition and the occurrence of senescence or apoptosis as a reversal of the immortal phenotype. Many telomerase inhibitors seem to be most efficient when combined with conventional chemotherapeutics. There are some promising approaches that seem to circumvent the slow way of telomere shortening and induce fast apoptosis in treated tumor cells. It has been demonstrated that telomerase may be involved in triggering apoptosis, but the underlying molecular mechanism remains unclear. PMID- 12757980 TI - Telomerase as tumor marker. AB - Telomerase, a critical enzyme responsible for continuous cell growth, is repressed in most somatic cells except proliferating progenitor cells and activated lymphocytes, and activated in approximately 85% of human cancer tissues. Telomerase activity is a useful cancer-cell detecting marker in some types of cancers in which almost all cases show telomerase activation. In other types in which telomerase becomes upregulated according to tumor progression, it is a useful prognostic indicator. Detection of human telomerase reverse transcriptase (hTERT) mRNA or protein in various clinical samples is also applicable. However, careful attention should be paid to the false negative results due to the instability of this enzyme or hTERT mRNA and the existence of polymerase chain reaction inhibitors as well as the false-positive results due to the contamination by normal cells with telomerase activity. If these pitfalls are avoided, in situ detection of hTERT mRNA or protein will facilitate the reliability of telomerase as a tumor marker. PMID- 12757981 TI - The role of the TSC-22 (-396) A/G variant in the development of diabetic nephropathy. AB - TSC-22 is a leucine zipper transcriptional factor and expression of the TSC-22 gene is highly induced by TGF-beta treatment. We estimated the frequency of the 396 A/G polymorphism of the TSC-22 gene with an Alu I-Restriction fragment length polymorphism (RFLP) method in 498 Japanese subjects with type 2 diabetes mellitus. We also determined the promoter activity. The diabetic patients with the AA genotype had a significantly higher incidence of the diabetic nephropathy (vs. the AG genotype, P<0.05, odds ratio: 1.95; 95% confidence intervals 1.14 3.33). There was no significant difference in the promoter activity between the fragments with -396A and -396G. These findings suggest that the TSC-22 gene ( 396) A allele is associated with an increasing risk of the diabetic nephropathy. PMID- 12757982 TI - Metabolic syndrome in urban Asian Indian adults--a population study using modified ATP III criteria. AB - AIMS/HYPOTHESIS: To determine the prevalence of the Metabolic syndrome (MetS) using modified ATP III criteria in urban Asian Indian adults. METHODS: 475 subjects (age 20-75 years) from a population data base were studied for the MetS using ATP III criteria but with a modified waist circumference (WC) appropriate for Indians. Presence of >or=3 of the following; raised WC (Men >or=90 cm, Women >or=85 cm), triglycerides (TG) >or=1.7 mmol/l), HDL-Cholesterol (HDL-C)-<1.0 mmol/l for men, <1.3 mmol/l for women, fasting plasma glucose (FPG)>or=6.1 mmol/l and blood pressure (BP)>or=130/>or=85 mm of Hg, or using BP medication, indicated the MetS. Insulin resistance (IR) was calculated using the Homeostasis Model Assessment (HOMA) equation. Factor analysis was used to identify clusters of correlated abnormalities. RESULTS: MetS was present in 41.1%. WC was increased in 31.4%, TG in 45.6%, low HDL-C in 65.5%, hypertension in 55.4% and raised FPG 26.7%. MetS was present in 27.9% of subjects with FPG<6.1 mmol/l and its prevalence increased to >70% with higher FPG values. MetS was more common in women than in men (46.5 vs. 36.4%, chi(2)=4.6, P=0.03) and in older people. Four distinct clusters of abnormalities were identified with some gender variations. IR was more prevalent in MetS and was a component of two clusters but it was not a core component in factor analysis. CONCLUSIONS: MetS is common in Asian Indians. Its prevalence is age-related, and is more common in women. HOMA-IR or fasting plasma insulin was not a core component of the MetS. PMID- 12757983 TI - Relation between insulin resistance and hematological parameters in elderly Koreans-Southwest Seoul (SWS) Study. AB - In this study, we investigated the relation between insulin resistance and hematological parameters in elderly Koreans. This study included 1314 non diabetic subjects over the age of 60, selected from a cross-sectional study, which was conducted in 1999 in Seoul, Korea. We measured fasting and post-load 2 h plasma glucose, insulin levels, lipid profiles, anthropometric measures, and hematological parameters. The degree of insulin resistance was assessed using the homeostasis model assessment (HOMA). We found a correlation between insulin resistance and hemoglobin concentrations in non-smoking men (r=0.20, P=0.0186). In non-smoking women, insulin resistance correlated with hemoglobin (r=0.10, P=0.0017) and with white blood cell (WBC) count (r=0.15, P=0.001). Hemoglobin concentrations and WBC counts were also associated with other components of the insulin resistance syndrome such as body mass index, blood pressure, lipid profiles and fasting plasma insulin levels (surrogate for insulin resistance). Furthermore, the group in the upper quartile for insulin resistance showed higher hemoglobin concentrations and WBC counts than the lower quartile, independent of smoking status and serum iron concentrations. Using HOMA-IR as a dependent variable in a multiple regression analysis, age, body mass index (BMI), waist-to hip ratio (WHR), systolic blood pressure, HDL cholesterol, triglyceride, WBC count, hemoglobin, hematocrit and serum TIBC were significant. Our results provide support for a relation between insulin resistance/hyperinsulinemia and hematological parameters such as hemoglobin concentrations and WBC counts in elderly Koreans. This suggests that increased erythropoiesis and subclinical inflammation could be part of the metabolic syndrome in elderly Koreans. PMID- 12757986 TI - Stress management training as related to glycemic control and mood in adults with Type 1 diabetes mellitus. AB - Relationships between attending a stress management and relaxation-training program, glycemic control (HbA(1c)) and mood were examined in two randomised groups of 31 persons with Type 1 diabetes. The program involved group-education 2 h a week for 14 weeks. Whereas one group received the program, the other acted as a control group and received the program later. HbA(1c) was measured and subjects filled out a mood adjective checklist before the start of intervention and both 1 month and 1 year after completing it. In both groups, significant positive mood changes were obtained, but no significant changes in HbA(1c) values occurred. No significant relationship was found between measures of change in HbA(1c) and of changes in mood. For those attending the group-sessions less frequently, the HbA(1c) values were significantly worse on each of the three measurement occasions than the values of those attending more frequently. The effectiveness of the program, with its failure to improve glycemic control but enhancing the mood of participants, is discussed in terms of characteristics of the sample and various methodological issues as well as in comparison with results of similar studies involving Type 1 and Type 2 diabetes. PMID- 12757984 TI - Successful treatment of necrotizing fasciitis associated with diabetic nephropathy. AB - A 50-year-old woman with a 15-year history of type 2 diabetes mellitus was admitted to our hospital due to high fever and a skin lesion with severe pain, swelling and a sensation of heat in the right thigh. Laboratory examination showed elevated C-reactive protein (CRP), thrombocytopenia, nephrotic syndrome and renal dysfunction. Her blood glucose level had been well controlled. Streptococcus agalactiae was detected in both the skin lesion and blood culture, and pathological examination revealed neutrophil infiltration in the fascia and muscle layer. The patient was diagnosed with necrotizing fasciitis, septic shock and disseminated intravascular coagulation. A combination therapy of antibiotics and surgical debridement resulted in the improvement of symptoms as supported by laboratory findings, and the skin lesion also showed improvement. Although group A streptococcus is well known to be implicated in the pathogenesis of necrotizing fasciitis, only S. agalactiae, belonging to group B streptococcus, was isolated from the tissue and blood cultures in this case. Although this organism is not virulent and rarely causes a necrotizing fasciitis, both the superficial fascial layer and underlying muscle were affected in this case. There have been only a few reports of necrotizing fasciitis due to S. agalactiae in patients with diabetes mellitus. Although the blood glucose level was well controlled in our patient, this disease might be caused by other factors, including diminished sense of touch and pain, abnormality of microcirculation and hypogammaglobulinemia due to nephrotic syndrome. PMID- 12757987 TI - Insulin lispro is as effective as regular insulin in optimising metabolic control and preserving beta-cell function at onset of type 1 diabetes mellitus. AB - The aim of the study was to examine the effects of intensive insulin therapy using lispro on metabolic control, immunogenicity and beta-cell function of newly diagnosed type 1 diabetic subjects in comparison with intensive insulin therapy using regular insulin. An open study was conducted in 45 newly diagnosed type 1 diabetic subjects. Patients were randomly assigned to intensive insulin therapy using insulin lispro (lispro) (lispro, n=22; 22.8 years) or intensive insulin therapy using regular insulin (regular) (regular, n=23; 24.4 years): three to five injections of subcutaneous rapid-acting insulin before meals and Neutral Protamine Hagedorn (NPH) before dinner/bed-time. GAD, IA2, insulin antibodies, basal and stimulated plasma C-peptide and HbA(1c) were measured initially and at months 1, 4, 8 and 12. Daily blood glucose profiles tended to be lower in the lispro group, particularly values after breakfast, without reaching statistical significance. There were no differences in terms of HbA(1c) throughout the study. The proportion of subjects achieving an HbA(1c)<6% at the end of the study was similar in both groups (regular 73.9%, lispro 68.0%). The number of mild hypoglycemic episodes tended to be lower with lispro, but not significantly. beta Cell function was not significantly different in both groups. During follow-up there were no differences in antibodies, including IAAb. In summary, insulin lispro used in intensive insulin therapy is as effective as regular insulin in optimizing metabolic control and preserving beta-cell function at diagnosis of type 1 diabetes. PMID- 12757989 TI - Evaluation of physical fitness in patients with Type 2 diabetes mellitus. AB - PURPOSE: The aim of this study was to compare functional capacity in 30 Type 2 Diabetic patients with 30 healthy non-diabetic control subjects. METHODS: Physical fitness was evaluated using the "EUROFIT Physical Fitness Test Battery". This battery estimates body composition, cardiopulmonary, musculoskeletal and motor fitness. RESULTS: Percentage of body fat (PBF) was higher in the diabetic compared with control groups (P<0.05) although body mass index (BMI) was similar. Biceps and suprailiac skinfold thickness were also greater in the diabetic group (P<0.05). The 6-min walking distance and VO(2max) were significantly lower in the diabetic group (P<0.05). The diabetic patients had lower values of the single leg balance test with eyes opened and closed. Jump-stretch, handgrip and side-bending of trunk tests were also lower in the diabetic patients. CONCLUSION: Physical functional capacity is lower in Type 2 diabetic patients than in age-matched control subjects. PMID- 12757988 TI - Comparison of glycaemic control and cardiovascular risk profile in patients with type 2 diabetes during treatment with either repaglinide or metformin. AB - OBJECTIVE: To compare glycaemic control and cardiovascular risk profile in patients with type 2 diabetes following 12 months' treatment with either repaglinide or metformin. STUDY DESIGN AND METHODS: This was an open uncontrolled randomised study in n=112 patients with inadequately controlled type 2 diabetes not previously treated with oral hypoglycaemic agents. Patients beginning treatment with either repaglinide or metformin entered an 8-week titration period (to optimise dosage: repaglinide, 2-4 mg/day; metformin, 1500-2500 mg/day) followed by a 12-month treatment period. Glycaemic control and cardiovascular risk factors were determined at baseline and at the end of the treatment period. RESULTS: Mean (S.D.) final drug doses were 3 (+/-1) mg/day in the repaglinide group and 2000 (+/-500) mg/day in the metformin group. Significant improvements in glycaemic control [glycated haemoglobin, fasting and 2-h postprandial plasma glucose (PPG)] were demonstrated in both treatment groups. The decrease in PPG was significantly greater in the repaglinide group (P<0.05). During the treatment period, fasting plasma insulin (FPI) decreased significantly in both groups, more so with metformin (P<0.05). Two-hour postprandial plasma insulin (PPI) levels decreased only in the metformin group (P<0.05). Significant improvements between baseline and final visit were demonstrated in one or both groups in the following cardiovascular risk factors: total cholesterol, low-density lipoprotein cholesterol (LDL-C), triglycerides, plasminogen activator inhibitor, lipoprotein(a) and homocysteine. No changes were observed in high-density lipoprotein cholesterol (HDL-C), apolipoprotein A-I, apolipoprotein B, fibrinogen, body mass index (BMI) or blood pressure. CONCLUSIONS: The use of repaglinide or metformin in drug therapy-nai;ve patients with type 2 diabetes over a 12-month period is associated with improvements in both glycaemic control and cardiovascular risk profile. The latter cannot necessarily be attributed to the pharmacotherapy per se, but provides reassurance in the context of initiating oral hypoglycaemic drug therapy with these agents. PMID- 12757990 TI - Impaired fasting glucose and risk of diabetes in Taiwan: follow-up over 3 years. AB - The purpose of this paper was to examine the relationship between fasting glucose levels and development of diabetes among residents of Penghu, Taiwan. From July 1995 to June 1996, a population-based cohort study was conducted among residents aged >or=40 years on the island of Penghu, Taiwan. Of the 1601 surveyed, 1306 (81.6%) did not have diabetes. Six hundred of these 1306 persons were re-examined 3 years later. Participants with fasting plasma glucose (FPG) concentration <110 mg/dl (<6.1 mmol/l) were classified as normoglycemic, those with a glucose concentration of 110-126 mg/dl (6.1-7.0 mmol/l) had impaired fasting glucose (IFG), and those with a fasting glucose concentration of >or=126 mg/dl (7.0 mmol/l) were considered to have diabetes. During the 3-year follow-up, 4.3% of the total population (1.4% per year, 95% CI 0.9-1.9%) developed diabetes. Of those with IFG at baseline, 9.6% (3.2% per year, 95% CI 1.8-5.0%) progressed to diabetes, but only 2.5% (0.8% per year, 95% CI 0.4-1.2%) of normoglycemic people did so. The multivariate-adjusted odds ratio of developing diabetes was 4.4 (95% CI 1.9-10.6) for persons with IFG compared with those who were normoglycemic at baseline. Other significant predictors of progression to diabetes were higher waist-hip ratio (WHR), triglyceride and apolipoprotein B (apo B) levels. In this Asian Chinese population, IFG is a strong predictor of diabetes. The high rate of conversion from IFG to diabetes, combined with the previously observed high IFG prevalence, suggests future high prevalence rates of diabetes in Taiwan. PMID- 12757991 TI - The effects of local review on informed consent documents from a multicenter clinical trials consortium. AB - There is increasing controversy about the appropriate role of the local institutional review board in the review of multicenter clinical studies. We evaluated the effects of the local review process at 25 study sites on the consent forms from two studies of the Tuberculosis Trials Consortium, a multicenter trials group. Two independent reviewers classified all changes made in the centrally approved consent forms; a third reviewer evaluated those changes if the two initial reviewers disagreed. The median time to initial local approval was 104.5 days (range 31-346). There were no changes in the study protocols as a result of local review. Consent forms became longer and less readable after local review, with a mean increase in grade level of 0.9 (+/-0.9) reading grade levels (p<0.001). A median of 46.5 changes (range 3-160) were made in the centrally approved forms. Most changes (85.2%) involved altering wording without affecting meaning. Errors were commonly introduced (11.2% of changes), and 33 of 50 (66%) locally approved consent forms contained at least one error in protocol presentation or a required consent form element. Local approval of two multicenter clinical trials was time-consuming and resulted in many changes in centrally approved consent forms. These changes frequently decreased readability and introduced errors. PMID- 12757992 TI - Toward protecting the safety of participants in clinical trials. AB - It is a widely held belief that the current system of oversight of clinical research, particularly the means of assessing risks and minimizing harms to participants in clinical trials, could be improved. In particular, the system is inefficient with overemphasis on the monitoring ability of some groups such as research ethics review boards and investigators, underemphasis on others such as data monitoring committees (DMCs) and sponsors, confusion about responsibilities for safety and imperfect communication between these different groups. Research ethics review boards are not able to perform safety monitoring by review of individual adverse events and are often burdened by duplicative reviews of large multicenter studies. There are no standards for DMCs to ensure they can reliably identify safety issues. Sponsors may be overreliant on data audits and slow to disseminate safety data in a coherent summary. Investigators, their staffs and clinical sites may not fully appreciate all the nuances of good clinical practice or may be inattentive to the daily conduct of studies. Regulators, particularly those in the United States, have failed to completely harmonize their policies with each other or with international regulatory agencies. We recommend well designed monitoring plans for all studies that are appropriate to their scope and risk, more centralized review of large multisite studies and closer local scrutiny of single-institution studies. In addition, sponsors should pay greater attention to monitoring adverse events and keeping up-to-date databases or investigator's brochures emphasizing safety issues. A minimal standard of education or expertise in good clinical practice should be established for investigators, their staffs and research ethics review board members. DMC composition and functions should be standardized and regulations should be harmonized nationally and internationally. Finally, there should be a concerted effort to study the efficacy of various components of the system. PMID- 12757993 TI - Perceptions of equipoise are crucial to trial participation: a qualitative study of men in the ProtecT study. AB - Recruitment to trials is known to be difficult. Previous research suggests that a crucial factor may be participants' difficulty with the concept of randomization. This study explored patients' perceptions of randomization and reasons for consent or refusal to participate in the ProtecT study (a randomized trial of surgery, radiotherapy, and monitoring for localized prostate cancer). In-depth interviews were conducted with 21 men diagnosed with localized prostate cancer who were invited to participate in the ProtecT treatment trial. Interviewees were selected purposefully from three U.K. clinical centers to ensure the inclusion of similar proportions of those agreeing or refusing random treatment allocation in each of the treatment groups. Interviews explored men's recall and understanding of chance, comparison, and equipoise, and reasons for consent/refusal of randomization and acceptance/rejection of treatment allocation. Data were analyzed methodically using the techniques of constant comparison. Checking of coding and interpretation was assured by four experienced qualitative researchers. Recall and understanding of the major principles of the randomized design were good and were similar for "chance" and "comparison" between those who consented to and refused randomization. Clinical equipoise, however, caused difficulty. Almost all recalled and understood it, but those who found it acceptable tended to consent to randomization and those who could not accept it tended to refuse to participate. Belief in clinical equipoise was key to participants' consent to randomization. Ensuring patients understand and accept equipoise may thus increase their readiness to consent to participate in trials. A priority for future research is to focus on the provision and presentation of suitable and effective trial information, concentrating in particular on the neglected concept of clinical equipoise. PMID- 12757994 TI - Sample sizes for comparing means of two lifetime distributions with type II censored data: application in an aging intervention study. AB - Sample size determination is a very important part of planning for clinical trials. Most clinical trials do not follow all their subjects to the terminal event, resulting in censored observations. This article presents a method of computing sample sizes required to achieve adequate statistical power to compare the means of two lifetime distributions when both samples are subject to type II censoring. Our approach is based on the location-scale family of log-transformed lifetime distributions as compared to that based on the log-rank test and the family of proportional hazards. Specific applications to log-normal distribution and Weibull distribution are also discussed. PMID- 12757995 TI - Surgical quality assurance in the Ischemic Optic Neuropathy Decompression Trial (IONDT). AB - The purpose of this article is to report the methods and results of the surgical quality assurance program associated with the Ischemic Optic Neuropathy Decompression Trial (IONDT). A surgical quality assurance committee developed and implemented a quality assurance program for a randomized clinical trial requiring surgical intervention. A surgical technique questionnaire was administered at two times during the study course, and maintenance of surgeon certification required submission and approval of a masked videotape of an optic nerve sheath decompression surgery by each study surgeon. Surgical quality was assessed through completion of surgical report forms and standardized, masked review of operative notes. Rates of compliance and intra- and interreviewer agreement were assessed for each aspect of the program. Twenty-five of 32 surgeons (81%) successfully completed and maintained certification. Item agreement varied from 21-92% among reviewers of satisfactory videotapes and 22-89% on unsatisfactory videotapes. Intrarater agreement for videotape acceptability was 11 of 13 (85%), and for specific surgical steps, 147 of 182 (81%). Operative notes were submitted for 123 of 125 (98%) patients receiving surgery. Interrater agreement on individual items ranged from 73-100%. Classification of individual items was identical on first and second review for 1285 of 1344 (95.6%) items. Overall agreement for individual reviewers was 93.8-97.8%. We conclude that use of a small peer review committee, which developed and oversaw a quality assurance program, allowed for consistent certification and monitoring of surgical performance. This in turn increased the credibility of the IONDT results, which demonstrated no difference in outcome between operated and unoperated groups of patients. PMID- 12757996 TI - Crossing controls to treatment in repeated-measures trials. AB - For repeated-measures trials with continuous outcomes in which one group is designated as control and the other treatment, it is sometimes advantageous to add the treatment to the control group in the last periods of the design rather than to continue with control conditions throughout the repeated measures. Many circumstances preclude crossing patients who were initially administered a test treatment back to control conditions so that a full crossover design is not possible. Maintaining the treatment in the treatment group while crossing controls over to treatment in the last periods can result in greater efficiency in estimating a treatment effect that is constant over measures than continuing with control conditions. In contrast, crossing controls is less efficient for estimating a slope parameter for the treatment effect. Equations are provided for both treatment-effect models to determine loss or gain in efficiency from crossing controls. The information from the measures in which controls are crossed to treatment decreases variances of other estimators of interest, such as subgroup-by-treatment interaction effects, stepped or quadratic changes in treatment effect, and interpatient random treatment-effect variance and covariances. Exploration of various models for changes in treatment effect over measures showed that the efficiency of crossing controls depended on the model. This sensitivity to model means that the practical and statistical advantages of crossing controls over to treatment only sometimes outweigh those of the traditional continued-control design. PMID- 12757997 TI - Constructing a database of individual clinical trials for longitudinal analysis. AB - Individual patient data are often required to evaluate how patient-specific factors modify treatment effects. We describe our experience combining individual patient data from 1946 subjects in 11 randomized controlled trials evaluating the effect of angiotensin-enzyme converting (ACE) inhibitors for treating nondiabetic renal disease. We sought to confirm the results of our meta-analysis of group data on the efficacy of ACE inhibitors in slowing the progression of renal disease, as well as to determine whether any study or patient characteristics modified the beneficial effects of treatment. In particular, we wanted to find out if the mechanism of action of ACE inhibitors could be explained by adjusting for follow-up blood pressure and urine protein. Each trial site sent a database of multiple files and multiple records per patient containing longitudinal data of demographic, clinical, and medication variables to the data coordinating center. The databases were constructed in several different languages using different software packages with unique file formats and variable names. Over 4 years, we converted the data into a standardized database of more than 60,000 records. We overcame a variety of problems including inconsistent protocols for measurement of key variables; varying definitions of the baseline time; varying follow-up times and intervals; differing medication-reporting protocols; missing variables; incomplete, missing, and implausible data values; and concealment of key data in text fields. We discovered that it was easier and more informative to request computerized data files and merge them ourselves than to ask the investigators to abstract partial data from their files. Although combining longitudinal data from different trials based on different protocols in different languages is complex, costly, and time-intensive, analyses based on individual patient data are extremely informative. Funding agencies must be encouraged to provide support to collaborative groups combining databases. PMID- 12757998 TI - Do arrhythmia patients improve survival by participating in randomized clinical trials? Observations from the Cardiac Arrhythmia Suppression Trial (CAST)and the Antiarrhythmics Versus Implantable Defibrillators Trial (AVID). AB - It is debatable whether patients benefit directly from participation in a randomized clinical trial. We attempt to address this question for participants in the Cardiac Arrhythmia Suppression Trial (CAST) and the Antiarrhythmics Versus Implantable Defibrillators (AVID) studies. Survival rates were compared between eligible patients who enrolled in the trials and eligible patients who did not enroll, adjusting for baseline covariates. In CAST, despite that the active therapy was found to confer an almost threefold increased risk of death, survival was similar between the 3163 enrolled and the 1363 nonenrolled eligible patients. However, when patients were under study management, their risk of death was approximately 20% lower than when they left study management. In AVID, overall survival was similar between the 1016 enrolled and the 1246 nonenrolled eligible patients. However, mortality was substantially higher among patients not enrolled because the referring physician mandated the type of therapy. Overall these observational analyses suggest a net improvement in survival for the participants in these two trials. PMID- 12758000 TI - Influence of suspended solids on acute toxicity of carbofuran to Daphnia magna: I. Interactive effects. AB - This study explored the effects on Daphnia magna from exposure to the pesticide carbofuran in combination with stress from suspended solids exposure. Our objective was to assess whether suspended solids affects the toxicodynamic response of D. magna to carbofuran. A series of laboratory experiments was performed where animals were exposed to carbofuran concentrations ranging from 0 to 160 microg/l in combination with suspended solids concentrations ranging from 0 to 10000 mg/l. In the absence of suspended solids, effects of carbofuran were dose dependent and resulted in an EC(50) of 92 microg/l. Exposure to suspended solids, up to extreme levels that may be encountered in the environment and in the absence of carbofuran, showed no measurable toxicity. When D. magna were exposed to a constant carbofuran concentration, the numbers of affected organisms increased with increasing suspended solids concentrations. At a suspended solids concentration of 1000 mg/l, the EC(50) for carbofuran was reduced by half to 45 microg/l. The relationship between the toxicity of carbofuran (microg/l) and the concentration of suspended solids (mg/l) can be described with the following equation: carbofuran EC(50)=72 exp(-0.00014 [suspended solids]). An analysis of the data indicates that this relationship is consistent with a potentiated toxicity mechanism rather than an additive model. PMID- 12758001 TI - Influence of suspended solids on acute toxicity of carbofuran to Daphnia magna: II. An evaluation of potential interactive mechanisms. AB - It has been demonstrated that simultaneous exposure of Daphnia magna to suspended solids and a carbamate pesticide potentiates the toxic response to the pesticide. The toxicodynamics between these stressors were investigated to determine possible mechanisms of interaction. Three experimental series were conducted with D. magna to determine: the effect of food availability on carbofuran toxicity; the effect of food availability on jointly administered carbofuran and suspended solids; and changes in the magnitude of effects which can occur with suspended solids of different composition. These experiments demonstrated that both carbofuran toxicity and the joint toxicity of carbofuran and suspended solids to D. magna can be modulated by food availability. While it is clear that food dilution could contribute to energy stress, it appears likely that additional interactive processes also contribute to the observed synergism between carbofuran and suspended solids. Additionally, decomposed peat was shown to be less of a stressor to these pelagic invertebrates than inorganic subsoil. PMID- 12758002 TI - Effects of the pyrethroid insecticide, cypermethrin, on a freshwater community studied under field conditions. I. Direct and indirect effects on abundance measures of organisms at different trophic levels. AB - The effects of the pyrethroid insecticide cypermethrin on a natural freshwater community were studied in small in situ enclosures over an 11-day period. The experiment was conducted in a eutrophic lake using a regression design that included three untreated controls and a gradient of six unreplicated cypermethrin concentrations, ranging from 0.01 to 6.1 microg/l. This paper is the first in a series of two, and describes the fate of cypermethrin and its effects on the abundance of crustaceans, rotifers, protozoans (cilliates and heterotrophic nanoflagellates (HNF)) and bacteria and the biomass of periphytic and planktonic algae. The concentration of cypermethrin decreased quickly during the experiment, with a half-life of 48 h for the total and 25 h for the dissolved fractions of cypermethrin, respectively. Cypermethrin proved to be acutely toxic to crustaceans in enclosures receiving nominal cypermethrin concentrations of >/=0.13 microg/l. No Effect Concentration (NEC) and median Effect Concentration (EC(50)) for the total crustacean community and cladoceran and copepod subgroups ranged between 0.02-0.07 and 0.04-0.17 microg/l, respectively, with copepods being less sensitive than cladocerans. The abundance of rotifers, protozoans and bacteria and the chlorophyll-a concentration of planktonic and periphytic algae was significantly related to the concentration of cypermethrin. All groups proliferated within 2-7 days after the cypermethrin application in those enclosures where the abundance of crustaceans was seriously affected by cypermethrin (i.e. >/=0.13 microg/l). We hypothesise that the proliferation of rotifers, protozoans, bacteria and algae was due to a reduced grazer control from crustaceans and thereby mediated indirectly by cypermethrin. The results of this experiment provide knowledge on how an entire microplankton community may respond to pyrethroids in nature, and the indirect effects observed on the community clearly demonstrates the necessity of multispecies field experiments in ecotoxicological risk assessment. PMID- 12758003 TI - Effects of the pyrethroid insecticide cypermethrin on a freshwater community studied under field conditions. II. Direct and indirect effects on the species composition. AB - The effects of cypermethrin, a commonly used pyrethroid insecticide, were studied in small in situ enclosures situated in an eutrophic lake over an 11-day period. The experimental design used a regression principle that included three untreated controls and a gradient of six unreplicated cypermethrin concentrations, ranging from 0.01 to 6 microg/l. This paper is the second in a series of two and describes the effects on the species composition of the crustacean, rotifer, periphyton and phytoplankton communities. Multivariate ordination technique (redundancy analysis (RDA) combined with Monte Carlo permutation tests) showed that exposure to cypermethrin caused significant changes in the species composition of the communities. Changes in the structure of the communities were observed following exposure to a nominal concentration of 0.13 microg cypermethrin per litre above. The direct acute effect of exposure to cypermethrin was a rapid decrease of many species of crustacean zooplankton. The alterations in crustacean species composition were probably due to variations in susceptibility to the direct toxic effects of cypermethrin. No effects concentration (NEC) for individual zooplankton species were calculated using inverse regression and revealed that copepod nauplii were the most sensitive (NEC=0.01 microg/l) of the crustacean groups examined. The observed alterations of the species composition of the autotrophic communities as well as of the rotifers were most likely caused indirectly by cypermethrin, mediated through the direct negative effects of the insecticide on the crustacean grazers. The results of this experiment provide further knowledge about the direct and indirect effects of pesticide stress on the ecosystem level. They also show that there is a variation in sensitivity between different species of zooplankton under natural conditions and thus exemplify the necessity of multispecies approaches in the risk assessment of pesticides. PMID- 12758004 TI - Effects of the antiandrogens, vinclozolin and cyproterone acetate on gonadal development in the Japanese medaka (Oryzias latipes). AB - This study was focused on determining the effects of exposure to antiandrogens on the gonadal development of Japanese medaka (Oryzias latipes). Test compounds included the fungicide, vinclozolin and the clinical antiandrogen, cyproterone acetate. Newly hatched medaka were exposed to aqueous solutions of vinclozolin (2500 microg/l) and the vinclozolin fungicide formulation, Ronilan (1000 and 5000 microg/l) and cyproterone acetate (1 and 10 microg/l), for 3 months. Histological evaluation of the gonadal tissues of exposed fish indicated that the 5000 microg/l concentration of the vinclozolin formulation (Ronilan) induced a low incidence of intersex (i.e. testis-ova) and the 2500 microg/l concentration of vinclozolin-affected spermatogenesis in males. Also, the vinclozolin treatments induced moderate ovarian atresia. Cyproterone acetate also induced a low incidence of testis-ova, but in contrast to the vinclozolin treatment the amount of ovarian tissue in the testis-ova was equal to or greater than the amount of testicular tissue. In the cyproterone acetate treatments, both oogenesis and spermatogenesis were moderately inhibited at all test concentrations. The results of this study indicate that antiandrogens have the potential to alter testicular development and gametogenesis in fish. However, research is needed to determine the mechanisms by which antiandrogens affect fish. PMID- 12758005 TI - Influence of LAS on marine calanoid copepod population dynamics and potential reproduction. AB - The toxicity of linear alkyl benzene sulfonate (LAS) to marine invertebrates is well documented under laboratory conditions using single-species tests. It is less known how LAS affects natural populations of aquatic organisms. We hypothesised that LAS was more toxic to the calanoid copepod Acartia sp. under natural conditions than Acartia tonsa under cultured conditions in the laboratory. This hypothesis was checked by a direct comparison of LAS toxicity in single-species and model (mesocosm) studies. The acute and sublethal effects of LAS on the survival and egg production of laboratory reared A. tonsa were examined by standard test, i.e. incubation with LAS without food for 24-72 h. The LC(50) and EC(50) values averaged 1.23 and 0.74 mg l(-1) for survival and egg production, respectively. These values are comparable to previous reports. The effects of LAS on a natural copepod community were also investigated under in situ conditions. A series of seven mesocosms (holding approx. 3 m(3) of seawater each) was established with two mesocosms being controls without LAS and five mesocosms with increasing concentrations of LAS ranging from 0.1 to 6.5 mg l(-1) applied as a single dose. The indigenous copepod community, dominated by Acartia sp. and Centropages sp., responded clearly to LAS concentrations above 0.1 mg l( 1). The calculated no effect value was 0.14 mg LAS l(-1) (95% CI=0.08-1.82 mg LAS l(-1)) for the entire copepod community including all development stages after 24 h exposure. The increased sensitivity under in situ conditions was probably promoted by the suboptimal growth conditions, e.g. no saturated food concentration or inadequate nutritive values of the food. The amount of food expressed as chlorophyll concentration was low (around 2 microg chl. a l(-1)) but was not affected by LAS. It appeared that the naupliar stages of Acartia and Centropages were the least affected by LAS and that new cohorts were able to develop 15 days after the dosing with LAS. PMID- 12758006 TI - Developmental estrogenic exposure in zebrafish (Danio rerio): I. Effects on sex ratio and breeding success. AB - In vivo studies of fishes exposed to xenoestrogens have reported vitellogenin (Vtg) induction, ovatestes, altered sex ratios, and impaired reproductive capacity. The objective of this study was to determine concentration dependent effects of a weak estrogen receptor agonist, 4-nonylphenol (NP) and a potent estrogen receptor agonist, 17alpha-ethinylestradiol (EE) on sex ratios, gonad morphology, Vtg induction and breeding success in zebrafish (Danio rerio). Fish were exposed from 2 to 60 days post-hatch (dph) to NP (10, 30, or 100 microg/l nominal), EE (1, 10, or 100 ng/l nominal), or solvent control (acetone; 0.2% v/v) in a static-renewal system with replacement every 48 h. At 60 dph, 20 fish from each treatment were euthanized for histological examination of gonads and Western blotting for Vtg using pooled heart homogenates. Remaining fish were reared in clean water until adulthood (120 dph) for breeding studies. Due to high mortality in the 100 ng/l EE group, insufficient fish were available for analyses. The percentage of males at 60 dph changed from 45% (9/20) in solvent controls to 0% at 10 ng/l EE and 10% at 100 microg/l NP. A concentration dependent increase in the number of fish with undeveloped gonads at 60 dph was observed in the EE exposure group. Two fish with ovatestes were observed at 100 microg/l NP, while one was observed at 30 microg/l NP. Western blotting showed induction of Vtg at 30 and 100 microg/l NP and 10 ng/l EE. The sex ratios of adults determined at 160 dph revealed no significant departure from 1 male:1 female, suggesting that exposure of zebrafish to estrogenic chemicals during sexual differentiation and early gametogenesis did not irreversibly alter phenotypic sex. The condition factor of adult fish and ovo-somatic index of adult female fish were also unaffected by developmental exposure to NP or EE. Despite this, breeding trials conducted in adult fish from 120 to 160 dph revealed significant reductions in the percent of viable eggs, hatchability, and swim-up success at 10 ng/l EE and 100 microg/l NP. Our results suggest that functional reproductive capacity (breeding success) may be more sensitive than gross morphological endpoints (condition, ovo-somatic index, sex ratio) in adult zebrafish exposed to xenoestrogens during sexual differentiation and early gametogenesis. PMID- 12758007 TI - Developmental estrogenic exposure in zebrafish (Danio rerio): II. Histological evaluation of gametogenesis and organ toxicity. AB - Aquatic species can be exposed to endocrine disrupting chemicals (EDCs) in wastewater that often includes the weak estrogen, 4-nonylphenol (NP) and the potent estrogen, 17alpha-ethinylestradiol (EE). The goal of the present study was to determine concentration-dependent effects of developmental exposure to NP and EE on gametogenesis, as well as gonad, kidney and liver pathology using quantitative histological evaluation of hematoxylin/eosin-stained saggital sections of zebrafish (Danio rerio). The major finding of the present study was that exposure to NP (>/=100 microg/l nominal) and EE (>/=1 ng/l nominal) from 2 to 60 days post-hatch (dph) caused concentration-dependent suppression of gametogenesis in both male and female zebrafish. Severe kidney pathology was observed in 60 dph zebrafish, specifically glomerular dilation or degeneration, fibrosis, tubule enlargement and tubule necrosis, at a threshold of 10 ng/l EE. However, minor kidney histopathology indicated by increased pyknotic nuclei in kidney tubule and interstitial (hematopoietic) cells was detected at lower estrogenic exposures (>/=10 microg/l NP nominal) than delayed gametogenesis. Considering all histological parameters in the current study, the rank order of potency for pathological effects in 60 dph zebrafish was 10 ng/l EE>1 ng/l EE=100 microg/l NP>30 microg/l NP>10 microg/l NP10 (nominal concentrations). Zebrafish from the same cohort examined in the current study that had been placed in clean water from 60 to 300 dph had histologically normal testes and no kidney or liver histopathology. However, increased ovarian follicle atresia was detected at 300 dph in zebrafish exposed developmentally to 100 microg/l NP. Therefore, we conclude that functional rather than morphological changes may be more important for future evaluations of developmental exposure to estrogens in fish, and that negative effects in female rather than male gonads may contribute to prolonged breeding impairment. PMID- 12758008 TI - Utility of the TBARS assay in detecting oxidative stress in white sucker (Catostomus commersoni) populations exposed to pulp mill effluent. AB - Recent evidence indicates that contaminant-stimulated free radical production and resulting oxidative damage may be an important mechanism of toxicity in organisms exposed to water-borne contaminants. This study tested the hypothesis that increases in oxidative stress and associated biochemical alterations would be present in white sucker (Catostomus commersoni) living in an environment receiving pulp mill effluent. Consistent increases in liver TBARS and frequent increases in gonadal TBARS were observed immediately downstream of the pulp mill effluent discharge; observable effects were attenuated with distance until they were not significantly different from reference values. Increases detected with the TBARS assay were commensurate with increases detected using the lipid hydroperoxides assay, a mechanistically independent technique. Fish exposed to pulp mill effluent also exhibited significant increases in hepatic free iron and ascorbic acid and a reduced free radical scavenging capacity in the livers of fish downstream of the effluent discharge relative to reference fish. Increases in oxidative stress are not necessarily dependent on increases in lipid substrate or related to reductions in ascorbic acid. TBARS values similar to those observed in fish resident below pulp mill effluent discharges were observed in white sucker 2 h after intraperitoneal injection using 15 mg/kg body weight ferric nitrilotriacetate (Fe(3+) NTA) as a positive control. This study indicates oxidative stress could be a mechanism of toxicity in fish exposed to pulp mill effluent and demonstrates the utility of TBARS in delineating zones of exposure to pulp mill effluent. PMID- 12758009 TI - Synthesis, characterization and thermal studies of (Ni/Co) metal salts of hydrazine: potential initiatory compounds. AB - Nickel hydrazinium nitrate (NiHN) and cobalt hydrazinium nitrate (CoHN) were prepared by reacting their respective metal nitrates with hydrazine hydrate at 25 and 65 degrees C. The compounds were characterized by metal content and infrared (IR) spectroscopy. Differential thermal analysis (DTA) results suggest that the nickel complex is relatively more stable than the cobalt complex. The activation energy determined by DTA and ignition delay measurements corresponds to an energy of activation (E(a)) of 80+/-4 kJ/mol for NiHN and that of 150+/-8 kJ/mol for CoHN. Thermo gravimetry (TG) also revealed more rapid decomposition of NiHN than that of CoHN in the temperature region of 215-235 degrees C. High temperature Fourier transform-infrared (FT-IR) studies indicated rupture of the Ni-Co-N bond as the primary step in the thermolysis. As regards sensitivity to mechanical stimuli, NiHN was found to be less impact sensitive while CoHN exhibited less friction sensitivity. The study revealed that NiHN could be used alone as well as in combination with oxidizer/fuel as initiators depending upon the specific requirements. The effect of silver azide and glass on the sensitization of NiHN was also studied. CoHN appears to be an effective ballistic modifier in enhancing burning rates of composite propellants. PMID- 12758010 TI - Mechanical recycling of waste electric and electronic equipment: a review. AB - The production of electric and electronic equipment (EEE) is one of the fastest growing areas. This development has resulted in an increase of waste electric and electronic equipment (WEEE). In view of the environmental problems involved in the management of WEEE, many counties and organizations have drafted national legislation to improve the reuse, recycling and other forms of recovery of such wastes so as to reduce disposal. Recycling of WEEE is an important subject not only from the point of waste treatment but also from the recovery of valuable materials.WEEE is diverse and complex, in terms of materials and components makeup as well as the original equipment's manufacturing processes. Characterization of this waste stream is of paramount importance for developing a cost-effective and environmentally friendly recycling system. In this paper, the physical and particle properties of WEEE are presented. Selective disassembly, targeting on singling out hazardous and/or valuable components, is an indispensable process in the practice of recycling of WEEE. Disassembly process planning and innovation of disassembly facilities are most active research areas. Mechanical/physical processing, based on the characterization of WEEE, provides an alternative means of recovering valuable materials. Mechanical processes, such as screening, shape separation, magnetic separation, Eddy current separation, electrostatic separation, and jigging have been widely utilized in recycling industry. However, recycling of WEEE is only beginning. For maximum separation of materials, WEEE should be shredded to small, even fine particles, generally below 5 or 10mm. Therefore, a discussion of mechanical separation processes for fine particles is highlighted in this paper. Consumer electronic equipment (brown goods), such as television sets, video recorders, are most common. It is very costly to perform manual dismantling of those products, due to the fact that brown goods contain very low-grade precious metals and copper. It is expected that a mechanical recycling process will be developed for the upgrading of low metal content scraps. PMID- 12758011 TI - Experimental study of the remediation of atrazine contaminated soils through soil extraction and subsequent peroxidation. AB - This paper presents a feasibility study in the field of the remediation of soils contaminated with atrazine. Experimental tests were performed on an artificially contaminated synthetic soil. Atrazine was removed from the soil by flushing with an aqueous solution at 5 vol.% of ethanol. Experimental tests of evaporation and Fenton's oxidation on the extracted solution were then performed in order to transform atrazine into its oxidation products. Tests were performed in the presence of a peroxide excess the ratio between Fe(2+) and H(2)O(2) was 1:10. Peroxide was first added in order to reduce the consumption of hydroxyl radicals by their reaction with the excess of Fe(2+). The degradation mechanism of atrazine during oxidation with Fenton's reagent in the presence of ethanol was investigated. Results showed that due to the non selective nature of Fenton's reagent a high consumption of reagent was needed to achieve a significant atrazine oxidation from solutions at 4.5 vol.% of ethanol. While at a Fe(2+) concentration of 3mM atrazine practically disappeared from pure aqueous solutions within 2h, a degradation yield of only 28.1% was observed in the presence of ethanol even when Fe(2+) concentration was 15 mM. PMID- 12758012 TI - Simultaneous multi-element detection of metal ions bound to a Datura innoxia material. AB - An on-line detection scheme has been developed for the determination of metal ion affinities for binding to a plant-based substrate. This involves monitoring the effluent of a column packed with cell-wall fragments from the plant Datura innoxia for 27 different elements simultaneously by coupling the column to an ICP emission spectrometer. Previously accepted procedures for removing native metal ions from biological materials by washing the material with a pH 2 solution were found to be insufficient for this material. Measurable amounts of Na, Mg, Al, Ca, Mn, Fe, Ni, Cr, Zn, Cd, Pb, Ba, Sr, and Si were all detected in an effluent from the introduction of 1.0M HCl following washing the material in a pH 2 solution. Metal ion breakthrough curves for Cd(2+), Zn(2+), Ni(2+), Cu(2+), and Pb(2+) were found to exhibit an affinity order of Pb(2+)>Cu(2+)>>Zn(2+) congruent with Cd(2+)>Ni(2+) for an equimolar mixture of these metal ions. This configuration also enabled the displacement of metal ions to be detected as the breakthrough curve for a subsequent metal ion was monitored. Comparison of Ni and Zn binding indicates a simple ion exchange model is insufficient to explain sequential binding of these metal ions. PMID- 12758013 TI - Landfarming operation of oily sludge in arid region--human health risk assessment. AB - Landfarming is becoming one of the most preferred treatment technologies for oily sludge disposal in the Arabian Gulf region in general, and in the Kingdom of Saudi Arabia in particular. This technology is considered to be, economical, energy efficient, and environmentally friendly with minimal residue disposal problems. Application of this technology in the region is simply based on the studies conducted in the United States of America and Europe. There have hardly been any scientific studies conducted to evaluate performance of landfarming technology under arid conditions. Recently, detailed field experimental study has been conducted to evaluate the degradation process and health risk issues in landfarming under arid conditions. The study observed volatilization as the main process of hydrocarbon degradation, which can cause significantly high concentration of airborne volatile organic compounds (VOCs) in the atmosphere leading to serious human health risk to the onsite workers. It is particularly true in the early phase of the landfarming process (first 2 months from initial loading). This paper elaborates these findings in detail. PMID- 12758015 TI - Kinetic behaviour of Duolite ES 468 in the cosorption of non-ionic surfactant and copper(II). AB - Kinetic behaviour of the hydrogen form of Duolite ES 468 polyacrylic acid functionalised cation exchanger with respect to the sorption of non-ionic surfactant alkylmonoethers (ALM-10) and copper(II) has been investigated; kinetic curves have been obtained, using spectrophotometric determination for ALM-10 and complexometric one for copper(II). Kinetic coefficient (B), intraparticle diffusion coefficient (D; m(2)s(-1)) and overall rate constant (k(o); s(-1)) for non-ionic surfactant and copper(II) depend on the solution composition, pH and the maximum sorption at the equilibrium. On increasing the solution acidity from pH 5 to pH 3 a decrease in both D and the equilibrium sorption for copper(II) although an increase in D for ALM-10 is observed. The action of copper(II) results in an increase in both D for ALM-10 and the maximum sorption at the equilibrium, whereas the action of ALM-10 leads to a decrease in the corresponding parameters for copper(II). Hydrogen form of Duolite ES 468 polyacrylic acid-functionalised cation exchanger is suitable for the simultaneous removal of non-ionic surfactant and copper(II) from waste water. PMID- 12758014 TI - Removal characteristics of anionic metals by micellar-enhanced ultrafiltration. AB - Surfactant-based separation of Fe(CN)(6)(3-) and CrO(4)(2-) using regenerated cellulose membrane was studied in order to assess the potential of micellar enhanced ultrafiltration for the remediation of wastewater or groundwater polluted with ferriccyanide and chromate. In the ferriccyanide/octadecylamine acetate (ODA) and chromate/ODA systems, removal of ferriccyanide increased from 73 to 92% and to 98%, and that of chromate from 64 to 97% and to >99.9% as the molar ratio of ODA to ferriccyanide and to chromate increased from 1 to 2 and to 3, respectively. In the ferriccyanide/chromate/ODA system, while the removal of ferriccyanide increased from 62 to 72% and to 93%, the removal of chromate from 20 to 38% and to 68% as the molar ratio of ferriccyanide:chromate:ODA increased from 1:1:1 to 1:1:2 and to 1:1:4, respectively. With the molar ratio of 1:1:6, the removal was >99.9 and 98% for chromate and ferriccyanide, respectively. Ferriccyanide ions were more easily bound to ODA micelles because the binding power of ferriccyanide was greater than that of chromate. PMID- 12758016 TI - Study of mercury pollution near a thermometer factory using lichens and mosses. PMID- 12758017 TI - Interactions between earthworms and arsenic in the soil environment: a review. AB - Chemical pollution of the environment has become a major source of concern. In particular, many studies have investigated the impact of pollution on biota in the environment. Studies on metalliferous contaminated mine spoil wastes have shown that some soil organisms have the capability to become resistant to metal/metalloid toxicity. Earthworms are known to inhabit arsenic-rich metalliferous soils and, due to their intimate contact with the soil, in both the solid and aqueous phases, are likely to accumulate contaminants present in mine spoil. Earthworms that inhabit metalliferous contaminated soils must have developed mechanisms of resistance to the toxins found in these soils. The mechanisms of resistance are not fully understood; they may involve physiological adaptation (acclimation) or be genetic. This review discusses the relationships between earthworms and arsenic-rich mine spoil wastes, looking critically at resistance and possible mechanisms of resistance, in relation to soil edaphic factors and possible trophic transfer routes. PMID- 12758018 TI - Concurrent plant uptake of heavy metals and persistent organic pollutants from soil. PMID- 12758019 TI - Deposition to forests in Europe: most important factors influencing dry deposition and models used for generalisation. AB - Dry deposition of gases and particles to forests is influenced by factors influencing the turbulent transport, such as wind speed, tree height, canopy closure, LAI, etc. as well as by factors influencing surface condition, such as precipitation, relative humidity, global radiation, etc. In this paper, an overview of these factors is given and it is shown which are the most important determining temporal and spatial variation of dry deposition of sodium and sulphur. Furthermore, it is evaluated how well current deposition models are able to describe the temporal and spatial variation in dry deposition. It is concluded that the temporal variation is not modelled well enough, because of limited surface-wetness exchange parameterisations. The influence of forest characteristics are modelled reasonably well, provided enough data describing the forests and the spatial variation in concentration is available. For Europe these data are not available. The means to decrease the atmospheric deposition through forest management is discussed. PMID- 12758020 TI - The annual course of TCA formation in the lower troposphere: a modeling study. AB - We present a modeling study investigating the influence of climate conditions and solar radiation intensity on gas-phase trichloroacetic acid (TCA) formation. As part of the ECCA-project (Ecotoxicological Risk in the Caspian Catchment Area), this modeling study uses climate data specific for the two individual climate regimes, namely "Kalmykia" and "Kola Peninsula". A third regime has also been included in this study, namely "Central Europe", which serves as a reference to somehow more moderate climate conditions. The simulations have been performed with a box modeling package (SBOX, photoRACM), which uses Regional Atmospheric Chemistry Mechanism (RACM) as its chemistry scheme. For this model a mechanism supplement has been developed including the reaction pathways of methyl chloroform photooxidation. The investigations are completed by a detailed sensitivity study addressing the impact of temperature and relative humidity. Atmospheric OH and HO2 concentrations and the NOx/HO2 ratio were identified as the governing quantities controlling the TCA formation trough methyl chloroform oxidation in the gas phase. Model calculations show a TCA production rate ranging between almost zero and 6.5 x 10(3) molecules cm(-3) day(-1) depending on location and season. In the Kalmykia regime the model predicts mean TCA production rates of 1.3 x 10(-4) and 5.4 x 10(-5) microg m(-3) year(-1) for the urban and rural environment, respectively. From the comparison of model calculations with measured TCA burdens in the soil ranging between 130 g m(-3) and 1750 g m(-3) we conclude that TCA formation through methyl chloroform photooxidation in the gas-phase is probably not the principal atmospheric TCA source in this region. PMID- 12758021 TI - Cu, Cr and As distribution in soils adjacent to pressure-treated decks, fences and poles. AB - Chromated copper arsenate (CCA)-treated wood has been widely used in the Southeastern United States to protect wood products from microbial and fungal decay. The aims of this study were to (1). determine the distribution of arsenic (As), chromium (Cr), and copper (Cu), in soils surrounding CCA-treated wood structures such as decks, fences and poles; and (2). evaluate the impacts of these structures on As, Cr and Cu loading of the soils. Profile and lateral soil samples were collected under CCA-treated decks and adjacent to poles and fences. The results showed elevation of As, Cr and Cu concentrations close to and under the structures, with mean As concentrations as high as 23 mg x kg(-1) close to utility poles compared with less than 3 mg x kg (-1) at distances of about 1.5 m away. Concentrations of As, Cr, and Cu decreased with depth in areas close to CCA treated poles. However, these results were only apparent in relatively new structures. A combination of weathering and leaching with time may have reduced the impact in older poles. Increased concentrations of As, Cu and Cr were also observed close to CCA-treated decks and fences, with age showing a similar impact. These results are helpful for CCA-treated wood product users to determine the safe use of these structures. PMID- 12758022 TI - The response of some common Egyptian plants to ozone and their use as biomonitors. AB - Relative sensitivity of five common Egyptian plant species namely, Senecio vulgaris, Malva parviflora, Sonchus oleraceus, Medicago sativa and Melilotus indicus to elevated levels of ozone has been studied. The plants were exposed to charcoal filtered air (CFA) and different levels of O3 (50 and 100 ppb) for 5 h per day. The studied parameters were recorded for five consecutive days after fumigation. The foliar injury varied significantly among species in a dose dependent manner. Severe injury symptoms were recorded on the leaves of M. sativa. With the exception of M. parviflora, all species exhibited significant increases in the percentage reduction of the above-ground dry weight as a result of reductions in both leaf and stem dry weights. M. sativa showed a marked reduction in its relative growth rate at elevated levels of O3. The extent of chlorophyll a destruction was higher in both M. sativa and S. oleraceus than in the other species tested. No differences in the sensitivity of chlorophylls a+b and carotenoids to ozone levels were recorded in this work. Percentage reduction of ascorbic acid was higher in M. sativa and S. oleraceus, compared with the other species studied. With respect to relative percentages of proline, there was a significant difference in the responses of plants to ozone. According to the ozone resistance (R%), measured as relative growth rate, the test species were arranged in the descending order: M. parviflora>M. Indicus>S. Vulgaris>S. Oleraceus>M. sativa. In M. sativa, both determinant and correlation coefficients are well reflected in the relationship between its physiological response, its performance and ozone levels, supporting its recommendation as a candidate for biomonitoring in Egypt. PMID- 12758023 TI - Variation of heavy metals within and among feathers of birds of prey: effects of molt and external contamination. AB - In this study we examined the effect of external contamination on the heavy metal (Ag, Al, As, Cd, Co, Cr, Cu, Fe, Hg, Mn, Ni, Pb and Zn) concentration in feathers. We compared the heavy metal content among the 10 primary wing feathers of sparrowhawks (Accipiter nisus), little owls (Athene nocta) and barn owls (Tyto alba) and the variation within the outermost tail feather of sparrowhawks and tawny owls (Strix aluco). The concentration of Hg was significantly higher in feathers molted first, suggesting that levels in feathers reflect levels in the blood during formation. For some other elements (Al, Co, Ni, Pb, Zn) on the other hand, there are strong indications that external contamination may have an important impact on the levels detected in the feathers. This should be taken into account in future monitoring studies. PMID- 12758024 TI - Effects of geographical location and land use on atmospheric deposition of nitrogen in the State of Connecticut. AB - A network of eight monitoring stations was established to study the atmospheric nitrogen concentration and deposition in the State of Connecticut. The stations were classified into urban, rural, coastal and inland categories to represent the geographical location and land use characteristics surrounding the monitoring sites. Nitrogen species including nitrate, ammonium, nitric acid vapor and organic nitrogen in the air and precipitation were collected, analyzed and used to infer nitrogen concentrations and dry and wet deposition flux densities for the sampling period from 1997 through 1999, with independently collected meteorological data. Statistical analyses were conducted to evaluate the spatial variations of atmospheric concentration and deposition fluxes of total nitrogen in Connecticut. A slightly higher atmospheric concentration of total nitrogen was observed along the Connecticut coastline of Long Island Sound compared to inland areas, while the differences of nitrogen deposition fluxes were insignificant between coastal and inland sites. The land use characteristics surrounding the monitoring sites had profound effects on the atmospheric nitrogen concentration and dry deposition flux. The ambient nitrogen concentration over the four urban sites was averaged 38.9% higher than that over the rural sites, resulting a 58.0% higher dry deposition flux in these sites compared to their rural counterparts. The local industrial activities and traffic emissions of nitrogen at urban areas had significant effects on the spatial distribution of atmospheric nitrogen concentration and dry deposition flux in the State. Wet and total deposition fluxes appeared to be invariant between the monitoring sites, except for high flux densities measured at Old Greenwich, a monitoring station near to and downwind of the New York and New Jersey industrial complexes. PMID- 12758025 TI - Genetic structure along a gaseous organic pollution gradient: a case study with Poa annua L. AB - The population genetic composition of Poa annua L. was studied by starch electrophoresis along a transect running NE from an organic reagents factory at Shanghai, China. Five enzyme systems were stained. We have reached the following preliminary conclusions: (1). Organic pollution has dramatically changed genotypic frequencies at some loci of Poa annua populations. At polluted sites, significant deviations from Hardy-Weinberg equilibrium were observed on loci Sod 1 and Me due to the excess of heterozygote. Especially in the two nearest sites to pollution source, all the individuals were heterozygous at locus Sod-1. The data suggests that heterozygotes were more tolerant to organic pollution than homozygotes, indicating the fitness superiority of heterozygotes. (2). A tendency towards clinal changes of allele frequencies was found at some polymorphic loci. Frequencies of the common alleles at loci Sod-1, Me and Fe-1 increased as the distance to the pollution source increased. (3). The effective number of alleles per locus, and the observed and expected heterozygosity were much higher in the pollution series than in the clear control site (Botanic Park population), but genetic multiplicity (number of alleles per locus) was lower than for the control. (4). Most genetic variability was found within populations, and only 2.56% were among populations of the polluted series. However, 9.48% of the total genetic variation occurred among populations when including the Botanic Park population. The genetic identity between populations of the pollution series (0.9869-1.0000, mean 0.9941) was higher than those between the pollution series and the Botanic Park population. UPGMA divided the five populations into two groups. One contained the four polluted populations, and the other only contained the Botanic Park population. PMID- 12758026 TI - Soluble and insoluble fractions of heavy metals in wet and dry atmospheric depositions in Bologna, Italy. AB - Atmospheric depositions were collected monthly using a modified wet and dry sampler (dry deposition was collected on a water surface) located in Bologna, a northern Italian urban area, to evaluate the impact of airborne heavy metals on the local pollution load. Wet deposition samples were filtered and heavy metal contents in soluble and insoluble fractions were determined. The same procedure was applied to the water samples which collected dry deposition. The entire procedure was tested using a certified reference material (CRM), which provided satisfying recovery results. The percentage of heavy metal soluble fraction in dry deposition was generally lower than in wet one; Cd, V, Cu and Zn showed a higher average solubility than Cr, Ni and Pb both in wet and dry deposition. Factor analysis, after a varimax rotation of principal components, suggested possible anthropogenic sources which explain different metal deposition patterns. This data analysis also allowed to distinguish different clusters formed by monthly fluxes of heavy metals. PMID- 12758027 TI - Evaluation of caged freshwater mussels as an alternative method for environmental effects monitoring (EEM) studies. AB - On three occasions between 1998 and 2000, freshwater mussels were collected by divers in Lake Memphremagog during the spring and transplanted to various locations in the St-Francois River (Quebec, Canada). Mussel growth was monitored by comparing total weight and length at the beginning and end of the exposure period. In 1998, mussels were caged for 60 days at 10 stations, including locations receiving treated effluents from three pulp and paper mills. Overall, there was an apparent trend of increased mussel growth from upstream to downstream along the river. However, mussels caged downstream from the effluent discharge of a bleached kraft pulp and paper mill grew more slowly than those caged immediately upstream in the river. In 1999 and 2000, we further investigated the situation in the vicinity of this bleached kraft mill. The measurements again indicated that growth of mussels in the effluent plume from this mill was reduced in comparison to sites upstream. Overall, in terms of growth, the caged mussels responded both positively and negatively to different environmental conditions. Compared with other monitoring approaches used at these sites during the same period, the caged mussel experiment results were consistent with the trends observed with the benthic invertebrate survey but not with the trends observed for fish. PMID- 12758028 TI - Impact of ozone on the growth of birch (Betula pendula) saplings. AB - Saplings of one half-sib family of birch, Betula pendula, were exposed to three levels of ozone in open-top chambers (OTCs) during two growing seasons 1997-1998. The ozone treatments were non-filtered air (NF, accumulated daylight AOT40 over the two growing seasons of 3.0 l l-1 h), non-filtered air with extra ozone (NF+, accumulated daylight AOT40 of 27.3 l l-1 h) and non-filtered air with additional extra ozone (NF++, accumulated daylight AOT40 of 120 l l-1 h). The birch saplings, including the roots, were harvested after the first and second growing seasons. After the first growing season, the NF++ treatment reduced the total wood biomass by 22%, relative to the NF treatment. There was no further reduction of the total wood biomass in the NF++ treatment after the second growing season. The root biomass was reduced by 30% after the first growing season. The shoot/root ratio, as well as the proportional biomass of leaves, were increased by ozone during both years. The ozone impact on the relative growth rate was estimated to -2% per 10 l l-1 h daylight AOT40 per growing season. PMID- 12758029 TI - Xenoestrogens in the River Elbe and its tributaries. AB - 4-Alkylphenols, 4-alkylphenol ethoxylates, 4-alkylphenoxy carboxylates, bisphenol A, bisphenol F, 4-hydroxyacetophenon, 4-hydroxybenzoic acid and steroid hormones were analyzed in water samples of the River Elbe and its tributaries Schwarze Elster, Mulde, Saale, Havel and Schwinge. Additionally, freshly deposited sediments (FDS, composite samples) of the River Elbe and its tributaries were analyzed. The concentrations in water samples ranged from (in ng/l): bisphenol A 4 to 92, branched nonylphenol 13 to 87, branched nonylphenol ethoxylates <0.5 to 120, 4-tert. nonylphenoxy carboxylates <10 to 940 and 4-hydroxybenzoic acid 4 to 12. Steroid hormones were only detected in the Czech tributaries Jizera and Vltava in concentrations near the limit of quantification. In FDS samples the concentrations amounted to (in g/kg d.w.): bisphenol A 10-380, branched nonylphenol 27-430, branched nonylphenol ethoxylates 24-3700, nonylphenoxy carboxylates <50 and 4-hydroxybenzoic acid 23-4400. Increased bisphenol A concentrations were found in water and FDS samples taken from the Czech-German border at Schmilka and the mouth of the Schwinge (only water sample). According to studies conducted in the Elbe Estuary and the German Bight, the River Elbe must be considered as a major source of pollution for the North Sea in respect of the compounds analyzed. A comparison of bisphenol A concentrations, 4 alkylphenols and the corresponding ethoxylates analyzed in the River Elbe and its tributaries with those found in other German surface waters indicated a low level of contamination. The evaluation of the data based on LOEC-values indicated that the concentrations were well below the effectivity threshold for some 4 alkylphenols. According to recent ecotoxicological investigations, for example, with prosobranch snails, bisphenol A concentrations found in water samples of the River Elbe and its tributaries may well be detrimental to aquatic organisms. On the basis of the monitoring data and its implications for estrogenic potency the inclusion of bisphenol A in the list of priority substances (European Union Directive 2000/60/EC, Annex X) should be considered. PMID- 12758030 TI - Trophic transfer of persistent organochlorine contaminants (OCs) within an Arctic marine food web from the southern Beaufort-Chukchi Seas. AB - Stable isotope values (13C, 15N) and concentrations of persistent organochlorine contaminants (OCs) were determined to evaluate the near-shore marine trophic status of biota and biomagnification of OCs from the southern Beaufort-Chukchi Seas (1999-2000) near Barrow, AK. The biota examined included zooplankton (Calanus spp.), fish species such as arctic cod (Boreogadus saida), arctic char (Salvelinus alpinus), pink salmon (Oncorhynchus gorbuscha), and fourhorn sculpin (Myoxocephalus quadricornis), along with marine mammals, including bowhead whales (Balaena mysticetus), beluga whales (Delphinapterus leucas), ringed seals (Phoca hispida) and bearded seals (Erignathus barbatus). The isotopically derived trophic position of biota from the Beaufort-Chukchi Seas marine food web, avian fauna excluded, is similar to other coastal food webs in the Arctic. Concentrations of OCs in marine mammals were significantly greater than in fish and corresponded with determined trophic level. In general, OCs with the greatest food web magnification factors (FWMFs) were those either formed due to biotransformation (e.g. p,p'-DDE, oxychlordane) or considered recalcitrant (e.g. HCH, 2,4,5-Cl substituted PCBs) in most biota, whereas concentrations of OCs that are considered to be readily eliminated (e.g. -HCH) did not correlate with trophic level. Differences in physical-chemical properties of OCs, feeding strategy and possible biotransformation were reflected in the variable biomagnification between fish and marine mammals. The FWMFs in the Beaufort Chukchi Seas region were consistent with reported values in the Canadian Arctic and temperate food webs, but were statistically different than FWMFs from the Barents and White Seas, indicating that the spatial variability of OC contamination in top-level marine Arctic predators is attributed to differences in regional sources of contamination rather than trophic position. PMID- 12758031 TI - Integrated assessment of the impacts of agricultural drainwater in the Salinas River (California, USA). AB - The Salinas River is the largest of the three rivers that drain into the Monterey Bay National Marine Sanctuary in central California. Large areas of this watershed are cultivated year-round in row crops and previous laboratory studies have demonstrated that acute toxicity of agricultural drainwater to Ceriodaphnia dubia is caused by the organophosphate (OP) pesticides chlorpyrifos and diazinon. In the current study, we used a combination of ecotoxicologic tools to investigate incidence of chemical contamination and toxicity in waters and sediments in the river downstream of a previously uncharacterized agricultural drainage creek system. Water column toxicity was investigated using a cladoceran C. dubia while sediment toxicity was investigated using an amphipod Hyalella azteca. Ecological impacts of drainwater were investigated using bioassessments of macroinvertebrate community structure. The results indicated that Salinas River water downstream of the agricultural drain is acutely toxic to Ceriodaphnia, and toxicity to this species was highly correlated with combined toxic units (TUs) of chlorpyrifos and diazinon. Laboratory tests were used to demonstrate that sediments in this system were acutely toxic to H. azteca, which is a resident genus. Macroinvertebrate community structure was moderately impacted downstream of the agricultural drain input. While the lowest macroinvertebrate abundances were measured at the station demonstrating the greatest water column and sediment toxicity and the highest concentrations of pesticides, macroinvertebrate metrics were more significantly correlated with bank vegetation cover than any other variable. Results of this study suggest that pesticide pollution is the likely cause of laboratory-measured toxicity in the Salinas River samples and that this factor may interact with other factors to impact the macroinvertebrate community in the system. PMID- 12758032 TI - Planning for remodelling: nuclear architecture, chromatin and chromosomes. AB - DNA sequences occupy three-dimensional positions and their architecture is related to gene expression, gene-protein interactions and epigenetic processes. The recent analysis of chromosome 4 in Arabidopsis interphase nuclei reveals that gene-rich, undermethylated DNA is composed of active loops of 200 to 2000 kb associated with acetylated histones, providing a well-defined model system to study chromatin in its nuclear context. PMID- 12758033 TI - Two genetically discrete pathways convert tryptophan to auxin: more redundancy in auxin biosynthesis. AB - The answer to the simple question of how plants make auxin has proven to be inordinately complex. Recent in planta studies in Arabidopsis have uncovered additional complexity in auxin biosynthesis. Two distinct pathways from tryptophan to the intermediate indoleacetaldoxime were identified. Genic, as well as functional redundancy, appear to be characteristic for auxin biosynthesis and plants might have evolved many different solutions for making and regulating auxin. PMID- 12758035 TI - Plants flex their skeletons. AB - Recent work on the fragile fiber mutants of Arabidopsis has identified microtubule-associated proteins that affect the orientation of cellulose microfibrils in cell walls, a major determinant of plant elongation growth. These same proteins are implicated in responses to gibberellin, provoking fresh speculation about how this hormone affects cell elongation and growth. PMID- 12758034 TI - Plants pass the salt. AB - Recently, overexpression of the plasma membrane Na(+)/H(+) antiporter SOS1 was shown to increase salt tolerance of Arabidopsis and revealed that levels of SOS1 mRNA are post-transcriptionally regulated by salt stress. In addition to demonstrating a novel approach to engineer salt-tolerant crops, the results provide the first glimpse of a previously unknown mechanism used by plants to regulate gene expression in response to salt stress. PMID- 12758036 TI - Insertional mutants: a foundation for assessing gene function. AB - The abundance of Arabidopsis insert mutants portends the day when null alleles in every gene will be obtained. Once these are created, all plant scientists can become geneticists. However, this brief technical highlight of genetic concepts cautions against ascribing gene function based exclusively on phenotypic analysis of null alleles. PMID- 12758037 TI - Risk assessment of GM plants: avoiding gridlock? AB - Cultivation of genetically modified crops is presently based largely on four crops containing few transgenes and grown in four countries. This will soon change and pose new challenges for risk assessment. A more structured approach that is as generic as possible is advocated to study consequences of gene flow. Hazards should be precisely defined and prioritized, with emphasis on quantifying elements of exposure. This requires coordinated effort between large, multidisciplinary research teams. PMID- 12758038 TI - ABA action and interactions in seeds. AB - The recent discovery of genes involved in abscisic acid (ABA) biosynthesis and responses heralds a new era for seed physiology. Our understanding of the regulation of ABA biosynthesis is moving from a linear metabolic pathway to a spatial and temporal network that governs ABA action in seeds. Transcription factors involved in ABA signaling have been identified, together with their target sequences. This allows further analysis of the specificity of ABA signaling in a complex system of interacting factors. PMID- 12758039 TI - 14-3-3 proteins find new partners in plant cell signalling. AB - 14-3-3 proteins are phosphoserine-binding proteins that regulate the activities of a wide array of targets via direct protein-protein interactions. In animal cells, the majority of their known targets are involved in signal transduction and transcription. In plants, we know about them primarily through their regulation of the plasma membrane H(+)-ATPase and enzymes of carbon and nitrogen metabolism. Nevertheless, an increasing number of plant signalling proteins are now being recognized as 14-3-3-interacting proteins. Plant 14-3-3 proteins bind a range of transcription factors and other signalling proteins, and have roles regulating plant development and stress responses. Important mechanisms of regulation by 14-3-3 include shuttling proteins between different cellular locations and acting as scaffolds for the assembly of larger signalling complexes. PMID- 12758040 TI - Green or red: what stops the traffic in the tetrapyrrole pathway? AB - Regulation of tetrapyrrole biosynthesis is crucial to plant metabolism. The two pivotal control points are formation of the initial precursor, 5-aminolaevulinic acid (ALA), and the metal-ion insertion step: chelation of Fe(2+) into protoporphyrin IX leads to haem and phytochromobilin, whereas insertion of Mg(2+) is the first step to chlorophyll. Recent studies with mutants and transgenic plants have demonstrated that perturbation of the branch point affects ALA formation. Moreover, one of the signals that controls the expression of genes for nuclear-encoded chloroplast proteins has been shown to be Mg-protoporphyrin-IX. Here, we discuss the regulation of branch-point flux and the relative contributions of the haem and chlorophyll branches to the regulation of ALA synthesis and thus to flow through the tetrapyrrole pathway. PMID- 12758041 TI - Making inroads into plant receptor kinase signalling pathways. AB - Cell-membrane-located receptor kinases play important roles in many plant signal transduction pathways. Exciting progress has been made in recent years with the characterization of four ligand-receptor systems involved in physiological processes as diverse as self-pollen rejection, stem-cell maintenance and differentiation at the shoot meristem, the response to the brassinosteroid hormones and the innate response to bacterial pathogens. These new findings emphasize the remarkably high diversity of these signalling pathways, although some downstream components are shared. This observation supports the idea that the wide diversification of plant receptors is associated with a high degree of specialization, one receptor potentially regulating a single developmental process. However, the possibility that one receptor might have a dual recognition function cannot be ruled out. PMID- 12758042 TI - Quorum quenching and proactive host defense. AB - Both plants and humans have inducible defense mechanisms. This passive defense strategy leaves the host unprotected for a period of time until resistance is activated. Moreover, many bacterial pathogens have evolved cell-cell communication (quorum-sensing) mechanisms to mount population-density-dependent attacks to overwhelm the host's defense responses. Several chemicals and enzymes have been investigated for years for their potential to target the key components of bacterial quorum-sensing systems. These quorum-quenching reagents, which block bacterial cell-cell communications, can disintegrate a bacterial population density-dependent attack. It has now been shown that a quorum-quenching mechanism can be engineered in plants and might be used as a strategy in controlling bacterial pathogens and to build up a proactive defense barrier. PMID- 12758043 TI - Respiratory tract infections. PMID- 12758044 TI - The classification of viruses infecting the respiratory tract. AB - Following the boom in respiratory virology in the 1960s, species of rhinoviruses, coronaviruses, enteroviruses, adenoviruses, parainfluenza viruses and respiratory syncytial virus were added to influenza and measles viruses as causes of respiratory tract infection. In restricted patient groups, such as the immunocompromised, members of the family of herpesviruses including herpes simplex, cytomegalovirus, varicella-zoster virus, Epstein-Barr virus and human herpes virus 6 have also been associated with respiratory disease. This list of pathogens was extended last year with the discovery of a novel virus, the human metapneumovirus. More than 200 antigenically distinct viruses have been documented as causes of sporadic or epidemic respiratory infections in infants, children and adults. However, this varied and diverse group can be divided among six distinct families. Understanding some of the basic biology of these families gives an insight into possible strategies for diagnosis, control and therapy. PMID- 12758045 TI - Human rhinoviruses. AB - Human rhinoviruses are the most important causative agents of upper respiratory infections and are also implicated in more severe clinical entities. Although often present, very little is known about human rhinoviruses. Molecular methods have been used in the classification of this large group of viruses into two separate clades. In addition, one known serotype was found to be a member of enterovirus group D. Laboratory diagnosis of human rhinovirus infection is based on reverse transcription polymerase chain reaction methods or the more tedious virus culture but a rapid "bedside" method is unavailable. Anti-rhinoviral therapy has been under extensive study over the past few decades but symptomatic treatment of the common cold is still the only useful approach in clinical use. More data on circulating human rhinovirus strains would facilitate both detection and treatment of these common pathogens. PMID- 12758046 TI - Influenza virus infection in infancy and early childhood. AB - Infants and young children have the highest influenza infection and hospitalisation rates in paediatrics. The immaturity of the infant's immune system and the absence of prior immunity and exposure to the virus are potential contributors. Although most children that suffer from influenza infection are otherwise healthy, an underlying chronic medical condition further increases the risk for complications. Annual immunisation with influenza vaccine is recommended for any child 6 months of age and older in whom prevention of disease is desirable, particularly for those with underlying medical conditions. Offering influenza vaccine to pregnant women who will deliver during the influenza season can potentially reduce the frequency and severity of influenza disease in infants less than 6 months of age. Family members, including other children and all other close contacts, should also receive influenza vaccine to reduce transmission to children at risk and infants in the first 6 months of life. PMID- 12758047 TI - Epidemiology and seasonality of respiratory tract virus infections in the tropics. AB - Acute viral respiratory tract infections are a significant cause of morbidity worldwide. Information on the epidemiology and seasonality of these infections is important in planning vaccination and treatment strategies. In temperate climes, there are distinct seasonal peaks in the winter months. This paper reviews the seasonal trends of respiratory viral infections in the tropics. Despite the absence of a winter season, consistent seasons of infection, albeit less distinct, have been observed. With few exceptions, respiratory syncytial virus and influenza infections have been observed mainly during the rainy seasons in Asian, African and South American countries. PMID- 12758048 TI - Immunology of viral respiratory tract infection in infancy. AB - Respiratory viruses such as respiratory syncytial virus (RSV), parainfluenza viruses (PIV) and the influenza viruses cause severe lower respiratory tract disease in infants and children throughout the world. We discuss the recent discovery of the epidemiologic importance of the human metapneumoviruses, first reported in 2001. Experimental live-attenuated vaccines for each of these viruses are being developed for intranasal administration in the first weeks or months of life. The immunology of these infections in humans is poorly defined but many studies are ongoing. A significant obstacle to successful immunisation of infants against respiratory-virus-associated disease early in life may be the relatively poor immune response of very young infants to primary virus infection. This paper reviews the immune correlates of protection against disease caused by these viruses, immune responses of infants to naturally acquired infection or experimental infection with candidate vaccine viruses and the genetics of susceptibility to severe disease. PMID- 12758049 TI - Growth factors in cystic fibrosis - when more is not enough. AB - In the airways of patients with cystic fibrosis, repeated cycles of infection and inflammation are responsible for bronchial wall thickening, a major determinant of loss of FEV(1) and progressive damage to the small and large airways. Proteolytic degradation of elastin, collagen and fibronectin fibrils in the tissue matrix leads to the loss of normal tissue architecture and the development of bronchiectasis, the most commonly observed morphological change on high resolution computed tomography examination. We have reviewed the evidence for increased expression of growth factors (TGF, HGF, FGF, EGF, VEGF) and activation of tissue repair processes in cystic fibrosis. Significantly higher concentrations of the growth factors compared with normal do not appear to prevent or reverse structural remodelling in the airways. The reasons why this process appears to be ineffective are discussed and we speculate on alternative strategies that might have a significant impact on the observed structural changes. PMID- 12758050 TI - Early sensitisation and development of allergic airway disease - risk factors and predictors. AB - The development and phenotypic expression of allergic airway disease depends on a complex interaction between genetic and several environmental factors, such as exposure to food, inhalant allergens and non-specific adjuvant factors (e.g. tobacco smoke, air pollution and infections). The first months of life seem to be a particularly vulnerable period and there is evidence that sensitisation is related to the level of allergen exposure during early life. At present, the combination of atopic heredity and elevated cord-blood IgE seems to result in the best predictive discrimination as regards development of allergic disease at birth. Early sensitisation, cow's milk allergy and atopic eczema are predictors for later development of allergic airway disease. Exposure to indoor allergens, especially house dust mite allergens, is a risk factor for sensitisation and development of asthma later in childhood in high-risk infants and infants with early atopic manifestations. PMID- 12758051 TI - Role of inhaler competence and contrivance in "difficult asthma". AB - Failure to deliver drug effectively to the lungs is a common cause of referrals with "difficult asthma". This may be due to poor regime compliance or poor device compliance (lack of competence and/or contrivance). The former is the more difficult to address. The latter can be addressed providing the healthcare professional is aware of the principles underlying aerosol delivery and aspects of patient behaviour. Unlike the gastrointestinal tract, the airways have evolved to exclude foreign material. A narrow window of opportunity exists with particles in the size range 1-7 micro m having a relatively high probability of depositing within the airways once inhaled. Current delivery systems are able to generate particles in this size range but they all have associated problems that may significantly impair the effectiveness of drug delivery to the lungs. While there is little evidence that compliance with inhaled therapy is significantly worse than with oral therapy, there is evidence that lack of competence (the inability to use a device effectively) or contrivance (knowing how to use a device effectively but contriving to use it ineffectively) are major additional impediments to effective therapy. Ensuring that the delivery system is acceptable to the patient is particularly important in the very young. The device chosen with a patient should be one that can and will be used effectively. For inhaled steroids, the choice of device/drug combination should be limited to those with a good therapeutic index (i.e. the majority of any systemic exposure should result from the lung dose). The best guide to determining the appropriate dose of inhaled steroid is to use the lowest effective dose. PMID- 12758052 TI - Bronchoscopy and infection. AB - Bronchoscopy can be useful to diagnose respiratory infections in the immunocompromised and those with problematic cystic fibrosis or with focally abnormal chest X-rays. Its role in paediatric tuberculosis is controversial with scanty objective evidence. As it is an invasive procedure, the following should be asked: what question am I trying to answer by bronchoscopy? Will the answer justify the risks of the procedure? PMID- 12758053 TI - Extracorporeal life support - state of the art. AB - Extracorporeal life support (ECLS) has become an accepted therapeutic measure in the treatment of infants, children and adults with reversible respiratory or cardiac failure. The principle behind ECLS involves obtaining access to drain blood from the venous circulation into the extracorporeal circuit where it is oxygenated and cleansed of carbon dioxide before being returned to the circulation. The UK Collaborative ECMO Trial showed that an ECLS policy was clinically effective in terms of improved survival without a rise in severe disability at age 1 year. Long-term follow-up has confirmed these benefits. The value of ECLS in paediatric and, more recently, adult respiratory failure is becoming clearer. ECLS has a vital role to play in the support of paediatric cardiac surgery programmes. Recent advances include newer oxygenators, greater use of less invasive veno-venous support and the use of ECLS to support novel therapies used to treat severe congenital diaphragmatic hernia. PMID- 12758054 TI - Inhaled glucocorticoids and adrenal function: an update. AB - For the vast majority of asthmatic children, treatment with inhaled glucocorticoids is safe and effective. Mild impairment of adrenal function of doubtful clinical significance is known to occur in some children inhaling > or = 400 micro g/day budesonide and beclomethasone or > or = 200 micro g fluticasone. Recent reports of life-threatening adrenal failure in asthmatic children inhaling glucocorticoids, some of whom were prescribed licensed doses, have prompted the recommendation that the use of high-dose inhaled glucocorticoids, particularly fluticasone, should be avoided. However, the importance of correctly diagnosing asthma, of using the minimum dose of inhaled glucocorticoid required for symptom control and of regular growth-velocity assessment cannot be over-emphasised. Appropriate asthma management including the early introduction of steroid-sparing agents such as a long-acting beta-agonist or leukotriene antagonist may reduce the morbidity associated with inhaled glucocorticoid use but some children, for reasons as yet unknown, may exhibit increased sensitivity to the systemic effects of inhaled glucocorticoid treatment. Possible explanations for this, with reference to the pharmacology and molecular mechanisms of glucocorticoid action, are accompanied in this review by a summary of the recent case reports and discussion of assessment of adrenal function. PMID- 12758056 TI - AhR- and ERK-dependent pathways function synergistically to mediate 2,3,7,8 tetrachlorodibenzo-p-dioxin suppression of peroxisome proliferator-activated receptor-gamma1 expression and subsequent adipocyte differentiation. AB - Activation of the aryl-hydrocarbon receptor (AhR) by pretreatment with 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD) blocks hormone (IDM/BRL)-induced adipocyte differentiation of C3H10T1/2 cells in proportion to the suppression of the elevation of the key mediator, peroxisome proliferator-activated receptor (PPARgamma1). Inhibition of MEK-induced ERK phosphorylation had no effect on adipogenesis but prevented this TCDD suppression. Initiation of MEK inhibition up to 6 h after IDM/BRL stimulation in combination with serum addition completely reversed the TCDD-mediated suppression but declined to ineffectiveness when delayed to 24 h after stimulation. This period occurs well after the decline of serum-induced ERK activation, at a time when ERK phosphorylation is low, and prior to the onset of IDM/BRL-stimulated PPARgamma1 expression. This temporal separation of ERK activation from the affected PPARgamma1 expression suggests that ERK does not act directly on either PPARgamma1 transcription or receptor function. Thus, ERK activation and TCDD/AhR stimulation work synergistically to inhibit adipocyte differentiation. Nonrenewal of serum at the time of IDM/BRL addition removed most of the ERK activation and also the TCDD-mediated suppressions of PPARgamma1 expression and adipocyte differentiation. Transfection of a vector expressing constitutively active MEK1 generated a constant, high level of phosphorylated ERK comparable to the peak serum-induced level and fully restored TCDD suppression without a TCDD-mediated effect on ERK phosphorylation. We conclude that low levels of activated MEK and ERK cooperate with AhR-induced factor(s) to generate a suppressor that prevents PPARgamma1 transcription and then differentiation. PMID- 12758055 TI - 2,2',4,6,6'-pentachlorobiphenyl (PCB 104) induces apoptosis of human microvascular endothelial cells through the caspase-dependent activation of CREB. AB - It has been proposed that endothelial integrity can play an active regulatory role in the extravasation of tumor cells during cancer metastasis. Since polychlorinated biphenyls (PCBs) have been shown to cause endothelial cell activation or injury and to lead to various diseases that involve dysfunction of the vascular endothelium, the present study was designed to determine the cellular and molecular signaling mechanisms of PCB-induced apoptosis in human microvascular endothelial cells (HMEC-1). A significant and marked decrease in cell viability was observed in HMEC-1 treated with 2,2',4,6,6' pentachlorobiphenyl (PCB 104) in a time- and dose-dependent manner. Exposure of HMEC-1 to PCB 104 also dramatically induced internucleosomal DNA fragmentation. However, the caspase inhibitor zVAD-fmk significantly reversed the PCB 104 induced DNA fragmentation in HMEC-1, suggesting that endothelial cell death induced by PCB 104 exposure is, at least in part, due to caspase-dependent apoptotic pathways. To elucidate the molecular signaling mechanisms of PCB 104 induced apoptotic cell death in human microvascular endothelial cells, the present study focused on the effects of acute exposure of PCB 104 on the activation of several transcription factors, such as cAMP responsive element binding protein (CREB), activator protein-1 (AP-1), nuclear factor-kappaB (NF kappaB), and signal transducers and activators of transcription (STAT1), which have been known to play a pivotal role in the molecular signaling cascades for the induction of apoptosis. A series of electrophoretic mobility shift assay showed that PCB 104 specifically increased only CREB DNA-binding activity in a dose-dependent manner. AP-1, NF-kappaB, and STAT1, however, were not activated. In addition, zVAD-fmk significantly and dose-dependently blocked the CREB activation enhanced by PCB 104 exposure. These results suggest that PCB-induced death of human microvascular endothelial cells is mediated, at least in part, via the caspase-dependent apoptotic pathways and that the selective activation of CREB is involved in this process. PMID- 12758057 TI - Experimental hepatic uroporphyria induced by the diphenyl-ether herbicide fomesafen in male DBA/2 mice. AB - Hepatic uroporphyria can be readily induced by a variety of treatments in mice of the C57BL strains, whereas DBA/2 mice are almost completely resistant. However, feeding of the protoporphyrinogen oxidase-inhibiting herbicide fomesafen (0.25% in the diet for 18 weeks) induced hepatic uroporphyria in male DBA/2N mice (liver porphyrin content up to 150 nmol/g, control animals 1 nmol/g), whereas fomesafen treated male C57BL/6N mice displayed only a slight elevation of liver porphyrins (approximately 5 nmol/g). The profile of accumulated hepatic porphyrins in fomesafen-treated DBA/2N mice resembled the well-characterised uroporphyria induced by polyhalogenated aromatic hydrocarbons, while histological examination confirmed the presence of uroporphyria-specific cytoplasmic inclusions in the hepatocytes. Uroporphyrinogen decarboxylase activity decreased to about 30% of control values in fomesafen-treated DBA/2N mice; microsomal methoxyresorufin O dealkylase activity was slightly reduced. The amount of CYP1A1 and CYP1A2 mRNA, as determined by real-time PCR, was not significantly changed; mRNA encoding the housekeeping 5-aminolevulinic acid synthase was elevated 10-fold. Total liver iron was slightly increased. A similar uroporphyria was induced by the herbicide formulation Blazer, containing a structurally related herbicide acifluorfen, when fed to DBA/2N mice at a dose corresponding to 0.25% of acifluorfen in the diet. Since DBA/2 mice are almost completely resistant to all well-characterised porphyrogenic chemicals, the results suggest the possible existence of a yet unknown mechanism of uroporphyria induction, to which the DBA/2 mouse strain is more sensitive than the C57BL strain. PMID- 12758058 TI - Oxidative DNA damage in vitamin C-supplemented guinea pigs after intratracheal instillation of diesel exhaust particles. AB - The health effects of diesel exhaust particles (DEP) are thought to involve oxidative damage. We have investigated the effect of intratracheal DEP instillation to guinea pigs in three groups of 12 animals each given 0, 0.7, or 2.1 mg. Five days later guinea pigs exposed to DEP had increased levels of oxidized amino acids (gamma-glutamyl semialdehyde), DNA strand breaks, and 7 hydro-8-oxo-2'-deoxyguanosine (8-oxodG) in the lung. Bulky DNA ad- ducts were not significantly elevated in the lung. The antioxidant enzyme activity of glutathione reductase was increased in the lung of DEP-exposed guinea pigs, whereas glutathione peroxidase and superoxide dismutase enzyme activities were unaltered. There was no difference in DNA strand breaks in lymphocytes or urinary excretion of 8-oxodG at the two doses tested. Protein oxidations in plasma and in erythrocytes were not altered by DEP exposure. The concentrations of ascorbate in liver, lung, and plasma were unaltered by the DEP exposure. The results indicate that in guinea pigs DEP causes oxidative DNA damage rather than bulky DNA adducts in the lung. Guinea pigs, which are similar to humans with respect to vitamin C metabolism, may serve as a new model for the study of oxidative damage induced by particulate matter. PMID- 12758059 TI - Suramin inhibits beta-bungarotoxin-induced activation of N-methyl-D-aspartate receptors and cytotoxicity in primary neurons. AB - We demonstrated that beta-bungarotoxin (beta-BuTX), a snake presynaptic neurotoxin, exhibited a potent cytotoxic effect on cultured cerebellar granule neurons. The mechanism of action of beta-BuTX and the cytoprotective agents against beta-BuTX were studied. The neuronal death of cerebellar granule neurons induced by beta-BuTX was manifested with apoptosis and necrosis processes as revealed by neurite fragmentation, morphological alterations, and staining apoptotic bodies with the fluorescent dye Hoechst 33258. By means of microspectrofluorimetry and fura-2, we measured intracellular Ca2+ concentration, [Ca2+]i and found that [Ca2+]i was increased markedly prior to the morphological changes and cytotoxicity. The downstream pathway of the increased [Ca2+]i was investigated: there was increased production of free radicals, decreased mitochondrial membrane potential, and depleted cellular ATP content. MK801 and suramin effectively suppressed these detrimental effects of beta-BuTX. Furthermore, the [3H]MK801 binding was reduced by unlabeled MK801, beta-BuTX, and suramin. Thus, activation of N-methyl-D-aspartate (NMDA) receptors appeared to play a crucial role in the cytotoxic effects following betaBuTX exposure. In conclusion, the novel finding of this study was that a polypeptide beta-BuTX exerted a potent cytotoxic effect through sequential events, including activating NMDA receptors followed by increasing [Ca2+]i, ROS production, and impaired mitochondrial energy metabolism. Suramin, clinically used as a trypanocidal agent, was an effective antagonist against beta-BuTX. Data suggest that suramin might have value to detect the possible pathway of certain neuropathological disorders. PMID- 12758060 TI - Different transport mechanisms for cadmium and mercury in Caco-2 cells: inhibition of Cd uptake by Hg without evidence for reciprocal effects. AB - Cadmium/Hg interactions have been studied in the TC7 clone of the enterocytic like Caco-2 cells to test the hypothesis that these metals may compete for intestinal transport. Comparison of the kinetic parameter values for 203Hg(II) and 109Cd(II) uptake in a serum-free medium revealed that Hg is accumulated much more rapidly and to higher concentrations. The very rapid uptake/binding step and the initial uptake rate of 109Cd were both significantly inhibited by an excess of unlabeled Cd or Hg (apparent K(i) for Hg of 9.3 +/- 1.2 microM) without reciprocal effects. 109cadmium uptake was highly sensitive to temperature and a significant fraction of accumulation (12%) was EDTA extractable. 203Hg uptake remained insensitive to temperature or the EDTA washing procedure. However, the uptake of both tracers was half-decreased when an excess of the respective unlabeled metal was added in the stop solution, suggesting an exchange mechanism for adsorption. Cell pretreatment with N-ethylmaleimide (NEM) led to a 30% decrease or a 73% increase in the 3-min specific transport of 109Cd when NEM was still present in or removed from the uptake medium, respectively. NEM had no effect on 203Hg uptake. Overall our results suggest the involvement of a saturable specific mechanism for Cd, which is highly sensitive to inhibition by Hg and NEM under some conditions, and a nonspecific passive diffusion for Hg. The Hg- or NEM-induced inhibition of Cd uptake likely involves a thiol-mediated reaction, but our results suggest that NEM pretreatment may activate other cellular mechanisms leading to a stimulatory effect. PMID- 12758061 TI - Impact of genetic profiles on experimental studies: outbred versus wild rats. AB - Most rats used in toxicological or pharmacological research are obtained from outbred stocks. It is assumed that they are characterized by a high degree of genetic heterogeneity, reflecting that of the human population. However, most outbred stocks available have been kept as restricted colonies over a long period of time, decreasing their genetic variability. Since it is unclear to what extent genetic variability can be expected from an outbred stock, we analyzed 20 rats from each of two outbred stocks, Crl:WIST and Rj:SD, and compared them with 17 unrelated wild rats, using 90 randomly selected microsatellite markers, located on 20 autosomes and chromosome X. Up to 8 alleles per locus were found in wild rats compared with the maximum of 4 alleles in Crl:WIST and 3 alleles in Rj:SD rats. Therefore, significant differences were observed in the mean of number of alleles between Crl:WIST (2.3 +/- 0.6) and Rj:SD (1.7 +/- 0.6), as well as between both outbred stocks and wild rats (4.6 +/- 1.5). The lowest heterozygosity was in Rj:SD (0.27 +/- 0.19), which was significantly different from Crl: WIST (0.41 +/- 0.19) and wild rats (0.35 +/- 0.21). Our findings clearly show loss of alleles in both outbred stocks compared with wild rats and demonstrate that the designation "outbred stock" is no guarantee for genetic and phenotypic variability. This fact will have a significant impact on generation of new outbred stocks as well as performing multistrain experiments and thereby it may reduce false-positive and false-negative results in toxicological and pharmacological research in the future. PMID- 12758062 TI - Regulation of brain function by exercise. AB - The effect of excercise on brain function was investigated through animal experiments. Exercise leads to increased serum calcium levels, and the calcium is transported to the brain. This in turn enhances brain dopamine synthesis through a calmodulin-dependent system, and increased dopamine levels regulate various brain functions. There are abnormally low levels of dopamine in the neostriatum and nucleus accumbens of epileptic mice (El mice strain) and spontaneously hypertensive rats (SHR). The low dopamine levels in those animals were improved following intracerebroventricular administration of calcium chloride. Dopamine levels and blood pressure in SHR were also normalized by exercise. In epileptic El mice, convulsions normalized dopamine levels and physiologic function. These findings suggest that exercise or convulsions affect brain function through calcium/calmodulin-dependent dopamine synthesis. This leads to the possibility that some symptoms of Parkinson's disease or senile dementia might be improved by exercise. PMID- 12758063 TI - Alzheimer disease-associated cystatin C variant undergoes impaired secretion. AB - CST3 is the coding gene for cystatin C (CysC). CST3 B/B homozygosity is associated with an increased risk of developing Alzheimer disease. We performed CysC analysis on human primary skin fibroblasts obtained from donors carrying A/A, A/B, and B/B CST3. Pulse-chase experiments demonstrated that the release of the B variant of CysC has a different temporal pattern compared to that of the A one. Fibroblasts B/B homozygous displayed a reduced secretion of CysC due to a less efficient cleavage of the signal peptide, as suggested by high-resolution Western blot analysis and by in vitro assay. In the brain, the reduced level of CysC may represent the molecular factor responsible for the increased risk of Alzheimer disease. PMID- 12758064 TI - Role of metallothionein-III following central nervous system damage. AB - We evaluated the physiological relevance of metallothionein-III (MT-III) in the central nervous system following damage caused by a focal cryolesion onto the cortex by studying Mt3-null mice. In normal mice, dramatic astrogliosis and microgliosis and T-cell infiltration were observed in the area surrounding the lesioned tissue, along with signs of increased oxidative stress and apoptosis. There was also significant upregulation of cytokines/growth factors such as tumor necrosis factor-alpha, interleukin (IL)-1 alpha/beta, and IL-6 as measured by ribonuclease protection assay. Mt3-null mice did not differ from control mice in these responses, in sharp contrast to results obtained in Mt1- Mt2-null mice. In contrast, Mt3-null mice showed increased expression of several neurotrophins as well as of the neuronal sprouting factor GAP-43. Thus, unlike MT-I and MT-II, MT III does not affect the inflammatory response elicited in the central nervous system by a cryoinjury, nor does it serve an important antioxidant role, but it may influence neuronal regeneration during the recovery process. PMID- 12758065 TI - Molecular investigation of TBP allele length: a SCA17 cellular model and population study. AB - Recently, an inherited spinocerebellar ataxia (SCA17) has been attributed to polyglutamine coding expansions within the gene coding for human TATA-box binding protein (TBP). The normal repeat range is 25-42 units with patients having as few as 46 repeats. We undertook a TBP repeat length population study showing its relative stability, skewed distribution, and substantial population specific differences. To investigate the mechanism of neurodegeneration in SCA17 we have developed a cellular model expressing full-length TBP with a range of polyQ expansions. As has been found with other polyQ cellular models, insoluble intracellular inclusions form in a repeat-length-dependent manner. In addition, we have shown that the expanded TBP polyQ tract is able to interact with other overexpressed polyQ-containing proteins. Importantly, overexpression of expanded TBP results in increased Cre-dependent transcriptional activity. As TBP is required for transcription by all RNA polymerases, this may indicate a mechanism for aberrant polyQ gain of function. PMID- 12758066 TI - Compensatory regulation of striatal neuropeptide gene expression occurs before changes in metabolic activity of basal ganglia nuclei. AB - Compensatory mechanisms delay the appearance of parkinsonian symptoms. However, both the order of appearance and potential interactions of compensatory mechanisms acting within the nigrostriatal pathway as well as inside and outside the basal ganglia are not clear. We hypothesize that, after the striatal dopaminergic homeostasis breakdown, a modification in the expression of several striatal markers (neuropeptide precursors and dopamine receptors) may occur before a change in the activity of both globus pallidus (GP) and substantia nigra pars reticulata (SNr) in response to a partial nigrostriatal lesion. The present data show, in MPTP-treated mice, that preproenkephalin-A and preprotachykinin mRNA expression and D(3) receptor binding are modified without changes in cytochrome oxidase metabolic activity in both GP and SNr, respectively. These changes in neuropeptide expression would compensate for the dopamine depletion induced changes in inhibitory GABAergic input from the striatum to GP and SNr. It also indicates that nondopaminergic compensatory mechanisms inherent to the basal ganglia are activated before those residing outside the basal ganglia. PMID- 12758067 TI - Age-dependent loss of PTP and LTP in the hippocampus of PrP-null mice. AB - We have investigated synaptic function in the hippocampus in mice of different ages carrying a null mutation in the PrP gene. Experiments carried out in vivo and in vitro in two laboratories revealed no differences in the ability of juvenile and young adult control and PrP-null mice to express long-term potentiation, paired-pulse facilitation, or posttetanic potentiation in either the dentate gyrus or in the CA1 region. However, we found a significant reduction in the level of posttetanic potentiation and long-term potentiation in the CA1 region of aged PrP-null mice. These results are discussed in relationship to reported increased levels of oxidative stress in older PrP-null mice. PMID- 12758068 TI - Potential use of early alterations in mu and delta opioid receptors as a predictive index for delayed brain ischemic damage. AB - We previously reported differential alterations of the mu, delta, and kappa opioid receptors following permanent middle cerebral artery occlusion. The present work studied the evolution of opioid receptor types following transient focal cerebral ischemia (tMCAO), as well as the putative predictive potential of early neurochemical alterations on the delayed ischemic damage. delta receptors were significantly decreased as early as 6 h post tMCAO (-22% approximately -57% vs. sham group), followed by a decrease in the mu binding site density at 24 h post tMCAO (-18% approximately -65%), in infarcted and penumbral cortices. Finally, early decreases in cortical opioid mu and delta receptor densities were found to significantly correlate (P < 0.001, r(2) = 0.48 and 0.75, respectively) with the occurrence of delayed histological damage. The high correlation between decreases in mu and delta receptor densities at 6 h post tMCAO and the histological damage that occurred at 24 h post tMCAO suggests that these early neurochemical alterations could be used as predictive markers of delayed ischemic damage. PMID- 12758069 TI - Reverse transcriptase and reverse splicing activities encoded by the mobile group II intron cobI1 of fission yeast mitochondrial DNA. AB - Mobile group II introns encode multidomain proteins with maturase activity involved in splicing and reverse transcriptase (RT) and (often) endonuclease activities involved in intron mobility. These activities are present in a ribonucleoprotein complex that contains the excised intron RNA and the intron encoded protein. Here, we report biochemical studies of the protein encoded by the group IIA1 intron in the cob gene of fission yeast Schizosaccharomyces pombe mitochondria (cobI1). RNP particle fractions from the wild-type fission yeast strain with cobI1 in its mtDNA have RT activity even without adding an exogenous primer. Characterization of the cDNA products of such reactions showed a strong preference for excised intron RNA as template. Two main regions for initiation of cDNA synthesis were mapped within the intron, one near the DIVa putative high affinity binding site for the intron-encoded protein and the other near domain VI. Adding exogenous primers complementary to cob exon 2 sequences near the intron/exon boundary stimulated RT activity but mainly for pre-mRNA rather than mRNA templates. Further in vitro experiments demonstrated that cobI1 RNA in RNP particle fractions can reverse splice into double-stranded DNA substrates containing the intron homing site. Target DNA primed reverse transcription was not detected unless a DNA target was used that was already nicked in the antisense strand of exon 2. This study shows that S.pombe cobI1 encodes RNP particles that have most of the biochemical activities needed for it to be a retroelement. Interestingly, it appears to lack an endonuclease activity, suggesting that the active homing exhibited by this intron in crosses may differ somewhat from that of the better-characterized introns. PMID- 12758071 TI - Teaching TetR to recognize a new inducer. AB - Tet Repressor (TetR) recognizes the inducer tetracycline (tc) with high affinity. The tc analog 4-de(dimethylamino)-6-deoxy-6-demethyl-tetracycline (cmt3) is not an inducer for TetR. Induction specificity for cmt3 was generated by employing a directed evolution approach to screen appropriate TetR mutants in four successive steps. The specificity of the best TetR mutant is more than 20,000-fold increased for cmt3 over tc as judged by the ratio of their respective binding constants. Two rounds of directed evolution via DNA shuffling revealed His64 as a key residue for inducer specificity. The best TetR mutant with cmt3 specificity contains the H64K exchange, leading to a 300-fold decreased tc and a 20-fold increased cmt3 affinity. Another round of directed evolution made use of randomized oligonucleotides to mutate selected residues close to the tc-binding pocket of TetR and yielded TetR S135L with a 250-fold increased cmt3 affinity. The double mutant TetR H64K S135L was constructed and again subjected to directed evolution using randomized oligonucleotides to alter residues in the "secondary shell" of the tc-binding pocket. The resulting best mutants TetR H64K E114Q S135L, TetR A61V H64K Q109E Q116E S135L and TetR H64K T112K S135L are fully inducible by cmt3 and not by tc. Thus, their inducer specificity has been redesigned. The molecular mechanism of changed inducer recognition is discussed, based on binding constants with several tc analogs and in light of the TetR crystal structure. PMID- 12758070 TI - Functional interplay between CBP and PCAF in acetylation and regulation of transcription factor KLF13 activity. AB - The transcriptional co-activators CBP/p300 and PCAF participate in transcriptional activation by many factors. We have shown that both CBP/p300 and PCAF stimulate the transcriptional activation by KLF13, a member of the KLF/Sp1 family, either individually or cooperatively. Here we further investigated how CBP and PCAF acetylation regulate KLF13 activity, and how these two co-activators functionally interplay in the regulation of KLF13 activity. We found that CBP and PCAF acetylated KLF13 at specific lysine residues in the zinc finger domain of KLF13. The acetylation by CBP, however, resulted in disruption of KLF13 DNA binding. Although the acetyltransferase activity of CBP is not required for stimulating the DNA binding activity of all of the transcription factors that we have examined, the disruption of factor DNA binding by CBP acetylation is factor specific. We further showed that PCAF and CBP act synergistically and antagonistically to regulate KLF13 DNA binding depending on the status of acetylation. PCAF blocked CBP acetylation and disruption of KLF13 DNA binding. Conversely, acetylation of KLF13 by CBP prevented PCAF stimulation of KLF13 DNA binding. PCAF blocked CBP disruption of KLF13 DNA binding by preventing CBP acetylation of KLF13. These results demonstrate that acetylation by CBP has distinct effects on transcription factor DNA binding, and that CBP and PCAF regulate each other functionally in their regulation of transcription factor DNA binding. PMID- 12758072 TI - Structural requirement for Mg2+ binding in the group I intron core. AB - Divalent metal ions are required for splicing of group I introns, but their role in maintaining the structure of the active site is still under investigation. Ribonuclease and hydroxyl radical footprinting of a small group I intron from Azoarcus pre-tRNA(Ile) showed that tertiary interactions between helical domains are stable in a variety of cations. Only Mg(2+), however, induced a conformational change in the intron core that correlates with self-splicing activity. Three metal ion binding sites in the catalytic core were identified by Tb(III)-dependent cleavage. Two of these are near bound substrates in a three dimensional model of the ribozyme. A third metal ion site is near an A minor motif in P3. In the pre-tRNA, Tb(3+) cleavage was redirected to the 5' and 3' splice sites, consistent with metal-dependent activation of splice site phosphodiesters. The results show that many counterions induce global folding, but organization of the group I active site is specifically linked to Mg(2+) binding at a few sites. PMID- 12758073 TI - Functionally distinct nucleic acid binding sites for a group I intron encoded RNA maturase/DNA homing endonuclease. AB - A large number of group I introns encode a family of homologous proteins that either promote intron splicing (maturases) or are site-specific DNA endonucleases that function in intron mobility (a process called "homing"). Genetic studies have shown that some of these proteins have both activities, yet how a single protein carries out both functions remains obscure. The similarity between respective DNA-binding sites and the RNA structure near the 5' and 3' splice sites has fueled speculation that such proteins may use analogous interactions to perform both functions. The Aspergillus nidulans mitochondrial COB group I intron encodes a bi-functional protein, I-AniI, that has both RNA maturase and site specific DNA endonuclease activities in vitro. Here, we show that I-AniI shows distinctive features of the endonuclease family to which it belongs, including highly specific, tight binding and sequential DNA strand cleavage. Competition experiments demonstrate that I-AniI binds the COB intron RNA even in saturating concentrations of its DNA target site substrate, suggesting that the protein has a separate binding site for RNA. In addition, we provide evidence that two different DNA-binding site mutants of I-AniI have little effect on the protein's RNA maturation activity. Since RNA splicing is likely a secondary adaptation of the protein, these observations support a model in which homing endonucleases may have developed maturase function by utilizing a hitherto "non-functional" protein surface. PMID- 12758074 TI - Flexible DNA target site recognition by divergent homing endonuclease isoschizomers I-CreI and I-MsoI. AB - Homing endonucleases are highly specific catalysts of DNA strand breaks that induce the transposition of mobile intervening sequences containing the endonuclease open reading frame. These enzymes recognize long DNA targets while tolerating individual sequence polymorphisms within those sites. Sequences of the homing endonucleases themselves diversify to a great extent after founding intron invasion events, generating highly divergent enzymes that recognize similar target sequences. Here, we visualize the mechanism of flexible DNA recognition and the pattern of structural divergence displayed by two homing endonuclease isoschizomers. We determined structures of I-CreI bound to two DNA target sites that differ at eight of 22 base-pairs, and the structure of an isoschizomer, I MsoI, bound to a nearly identical DNA target site. This study illustrates several principles governing promiscuous base-pair recognition by DNA-binding proteins, and demonstrates that the isoschizomers display strikingly different protein/DNA contacts. The structures allow us to determine the information content at individual positions in the binding site as a function of the distribution of direct and water-mediated contacts to nucleotide bases, and provide an evolutionary snapshot of endonucleases at an early stage of divergence in their target specificity. PMID- 12758075 TI - Thermophilic topoisomerase I on a single DNA molecule. AB - Control of DNA topology is critical in thermophilic organisms in which heightened ambient temperatures threaten the stability of the double helix. An important role in this control is played by topoisomerase I, a member of the type IA family of topoisomerases. We investigated the binding and activity of this topoisomerase from the hyperthermophilic bacterium Thermotoga maritima on duplex DNA using single molecule techniques, presenting it with various substrates such as (+) plectonemes, (-) plectonemes, and denaturation bubbles. We found the topoisomerase inactive on both types of plectonemes, but active on denaturation bubbles produced at increased stretching forces in underwound DNA. The relaxation rate depended sensitively on the applied force and the protein concentration. These observations could be understood in terms of a preference of the topoisomerase for single-stranded DNA over double-stranded DNA and allowed for a better understanding of activity of the topoisomerase in bulk experiments on circular plasmids. Binding experiments on a single duplex molecule using a mutant unable to perform cleavage confirmed this interpretation and suggested that T.maritima topoisomerase I behaves like an SSB by lowering the denaturation threshold of underwound DNA. Finally, experiments with a unique single-stranded DNA showed that both ends of the cleaved DNA are tightly maintained by the enzyme, supporting an enzyme-bridged mechanism for this topoisomerase. PMID- 12758076 TI - The 9.8 kDa subunit of complex I, related to bacterial Na(+)-translocating NADH dehydrogenases, is required for enzyme assembly and function in Neurospora crassa. AB - A nuclear gene encoding a 9.8 kDa subunit of complex I, the homologue of mammalian MWFE protein, was identified in the genome of Neurospora crassa. The gene was cloned and inactivated in vivo by the generation of repeat-induced point mutations. Fungal mutant strains lacking the 9.8 kDa polypeptide were subsequently isolated. Analyses of mitochondrial proteins from mutant nuo9.8 indicate that the membrane and peripheral arms of complex I fail to assemble. Respiration of mutant mitochondria on matrix NADH is rotenone-insensitive, confirming that the 9.8 kDa protein is required for the assembly and activity of complex I. We found a similarity between the MWFE homologues and the C-terminal part of the nqrA subunit of bacterial Na(+)-translocating NADH:quinone oxidoreductases (Na(+)-NQR), suggesting a link between proton-pumping and sodium pumping NADH dehydrogenases. PMID- 12758077 TI - A spatially extended stochastic model of the bacterial chemotaxis signalling pathway. AB - We have combined two distinct but related stochastic approaches to model the Escherichia coli chemotaxis pathway. Reactions involving cytosolic components of the pathway were assumed to obey the laws of conventional stochastic chemical kinetics, while the clustered membrane receptors were represented in two dimensional arrays similar to the Ising model. Receptors were assumed to flip between an active and an inactive state with probabilities dependent upon three energy inputs: ligand binding, methylation level due to adaptation, and the activity of neighbouring receptors. Examination of models with different lattice size and geometry showed that the sensitivity to stimuli increases with lattice size and the nearest-neighbour coupling strength up to a critical point, but this amplification was also accompanied by a proportional increase in steady-state noise. Multiple methylation of receptors resulted in diminished signal-to-noise ratio, but showed improved stability to variation in the coupling strength and increased gain. Under the best conditions the simulated output of a coupled lattice of receptors closely matched the time-course and amplitude found experimentally in living bacteria. The model also has some of the properties of a cellular automaton and shows an unexpected emergence of spatial patterns of methylation within the receptor lattice. PMID- 12758078 TI - Investigation of the role of the histidine-aspartate pair in the human exonuclease III-like abasic endonuclease, Ape1. AB - Hydrogen bonded histidine-aspartate (His-Asp) pairs are critical constituents in several key enzymatic reactions. To date, the role that these pairs play in catalysis is best understood in serine and trypsin-like proteases, where structural and biochemical NMR studies have revealed important pK(a) values and hydrogen bonding patterns within the catalytic pocket. However, the role of the His-Asp pair in metal-assisted catalysis is less clear. Here, we apply liquid state NMR to investigate the role of a critical histidine residue of apurinic endonuclease 1 (Ape1), a human DNA repair enzyme that cleaves adjacent to abasic sites in DNA using one or more divalent cations and an active-site His-Asp pair. The results of these studies suggest that the Ape1 His-Asp pair does not function as either a general base catalyst or a metal ligand. Rather, the pair likely stabilizes the pentavalent transition state necessary for phospho-transfer. PMID- 12758079 TI - Cooperative binding of dominant-negative prion protein to kringle domains. AB - Conversion of the cellular prion protein (PrP(C)) to the pathogenic isoform (PrP(Sc)) is a major biochemical alteration in the progression of prion disease. This conversion process is thought to require interaction between PrP(C) and an as yet unidentified auxiliary factor, provisionally designated protein X. In searching for protein X, we screened a phage display cDNA expression library constructed from prion-infected neuroblastoma (ScN2a) cells and identified a kringle protein domain using full-length recombinant mouse PrP (recMoPrP(23-231), hereafter recMoPrP) expressing a dominant-negative mutation at codon 218 (recMoPrP(Q218K)). In vitro binding analysis using ELISA verified specific interaction of recMoPrP to kringle domains (K(1+2+3)) with higher binding by recMoPrP(Q218K) than by full-length recMoPrP without the mutation. This interaction was confirmed by competitive binding analysis, in which the addition of either a specific anti-kringle antibody or L-lysine abolished the interaction. Biochemical studies of the interactions between K(1+2+3) and various concentrations of both recMoPrP molecules demonstrated binding in a dose dependent manner. A Hill plot analysis of the data indicates positive cooperative binding of both recMoPrP(Q218K) and recMoPrP to K(1+2+3) with stronger binding by recMoPrP(Q218K). Using full-length and an N-terminally truncated MoPrP(89-231), we demonstrate that N-terminal sequences enable PrP to bind strongly to K(1+2+3). Further characterization with truncated MoPrP(89-231) refolded in different conformations revealed that both alpha-helical and beta-sheet conformations bind to K(1+2+3). Our data demonstrate specific, high-affinity binding of a dominant negative PrP as well as binding of other PrPs to K(1+2+3). The relevance of such interactions during prion pathogenesis remains to be established. PMID- 12758081 TI - Sheared-type G(anti).C(syn) base-pair: a unique d(GXC) loop closure motif. AB - Stable DNA loop structures closed by a novel G.C base-pair have been determined for the single-residue d(GXC) loops (X=A, T, G or C) in low-salt solution by high resolution nuclear magnetic resonance (NMR) techniques. The closing G.C base-pair in these loops is not of the canonical Watson-Crick type, but adopts instead a unique sheared-type (trans Watson-Crick/sugar-edge) pairing, like those occurring in the sheared mismatched G.A or A.C base-pair, to draw the two opposite strands together. The cytidine residue in the closing base-pair is transformed into the rare syn domain to form two H-bonds with the guanine base and to prevent the steric clash between the G 2NH(2) and the C H-5 protons. Besides, the sugar pucker of the syn cytidine is still located in the regular C2'-endo domain, unlike the C3'-endo domain adopted for the pyrimidines of the out-of-alternation left-handed Z-DNA structure. The facile formation of the compact d(GXC) loops closed by a unique sheared-type G(anti).C(syn) base-pair demonstrates the great potential of the single-stranded d(GXC) triplet repeats to fold into stable hairpins. PMID- 12758080 TI - Leishmania mexicana glycerol-3-phosphate dehydrogenase showed conformational changes upon binding a bi-substrate adduct. AB - Certain pathogenic trypanosomatids are highly dependent on glycolysis for ATP production, and hence their glycolytic enzymes, including glycerol-3-phosphate dehydrogenase (GPDH), are considered attractive drug targets. The ternary complex structure of Leishmania mexicana GPDH (LmGPDH) with dihydroxyacetone phosphate (DHAP) and NAD(+) was determined to 1.9A resolution as a further step towards understanding this enzyme's mode of action. When compared with the apo and binary complex structures, the ternary complex structure shows an 11 degrees hinge bending motion of the C-terminal domain with respect to the N-terminal domain. In addition, residues in the C-terminal domain involved in catalysis or substrates binding show significant movements and a previously invisible five-residue loop region becomes well ordered and participates in NAD(+) binding. Unexpectedly, DHAP and NAD(+) appear to form a covalent bond, producing an adduct in the active site of LmGPDH. Modeling a ternary complex glycerol 3-phosphate (G3P) and NAD(+) with LmGPDH identified ten active site residues that are highly conserved among all GPDHs. Two lysine residues, Lys125 and Lys210, that are presumed to be critical in catalysis, were mutated resulting in greatly reduced catalytic activity. Comparison with other structurally related enzymes found by the program DALI suggested Lys210 as a key catalytic residue, which is located on a structurally conserved alpha-helix. From the results of site-directed mutagenesis, molecular modeling and comparison with related dehydrogenases, a catalytic mechanism of LmGPDH and a possible evolutionary scenario of this group of dehydrogenases are proposed. PMID- 12758082 TI - Regulation of RecA protein binding to DNA by opposing effects of ATP and ADP on inter-domain contacts: analysis by urea-induced unfolding of wild-type and C terminal truncated RecA. AB - RecA protein requires ATP and its hydrolysis to ADP to complete the DNA strand exchange reaction. We investigated how the nucleotides activate RecA by examining their effect on urea-induced unfolding, which could reflect domain-domain contact of protein. RecA is folded into three continuous domains: the N-terminal, central and C-terminal domains. The fluorescence of tyrosine residues, which lie mainly in the central domain, was modified in 1-3 M urea, while the red shift of fluorescence peak of the tryptophan residues located in the C-terminal domain occurred only in 3-6 M urea. Thus, the C-terminal domain of RecA is unfolded after the central part unfolds. The change in intensity of tryptophan fluorescence without a large shift in the peak at low concentrations of urea suggests that there are weak interactions between the central and C-terminal domains. This is supported by our observation that RecA protein lacking the C terminal tail unfolded at lower concentrations of urea than the entire RecA, and with clear transitions, unlike the entire RecA. ATP and its unhydrolyzable analog (ATPgammaS), which enhance the binding of RecA to DNA, facilitated the urea induced change in RecA tryptophan fluorescence, while ADP, an antagonist of ATP, prevented the change. ATP probably weakens the domain-domain contact and facilitates the DNA binding, while ADP stabilizes the contact and inhibits it. Supporting this conclusion, the binding of RecA lacking the C-terminal tail to DNA was not inhibited by ADP, while that of the intact RecA was. PMID- 12758083 TI - Can Forster resonance energy transfer measurements uniquely position troponin residues on the actin filament? A case study in multiple-acceptor FRET. AB - Straightforward interpretation of Forster resonance energy transfer (FRET) data in terms of the distance from donor-labeled troponon-tropomyosin (TnTm) to acceptor-labeled actin is complicated by the potential for energy transfer to acceptors on neighboring actin monomers (cross-transfer). Calculations indicate that cross-transfer can account for a substantial percentage of the total transfer efficiency. In some cases, this renders isolated FRET data uninterpretable. To overcome these limitations, we have developed an analysis method that incorporates cross-transfer and can, in principle, define the most probable (in the "least-squares" sense) position of a TnTm residue on the actin filament. The technique analyzes data from four or more FRET experiments using acceptors attached to different residues on actin. We have used this method to specify the coordinates of skeletal troponin I (sTnI) residue 133 relative to the actin filament under Mg(2+) and Ca(2+)-saturating conditions. Ca(2+)-activation causes the C terminus of the regulatory domain of TnI to move away from the actin surface by 6.3A, laterally along the actin surface toward actin subdomain 3 by 22.0A, and azimuthally toward the actin inner domain by 13.2A. This information is used to construct a low-resolution structural model of thin filament activation. PMID- 12758084 TI - Searching sequence space for high-affinity binding peptides using ribosome display. AB - We present the construction of a synthetic library based on the scaffold of bovine heart fatty acid-binding protein (FABP) with 1.1x10(14) independent members. Ribosome display was applied to select streptavidin-binding peptides in vitro from 2x10(13) molecules of the library each encoding FABP with 15 contiguous random amino acid residues at its N terminus. The selection yielded several different binding peptides. The best binder possessed a dissociation constant as low as 4nM and, in contrast to the previously isolated peptides, contained no HPQ motif. A substitution analysis enabled shortening of the 15-mer peptide and revealed a 9-mer variant with a dissociation constant of 17nM, which is a 1000-fold increase of affinity compared to the already known peptides of this size. This high-affinity binding peptide in combination with the whole set of streptavidin conjugates should be an extremely useful tool for the detection and purification of recombinant proteins. PMID- 12758086 TI - Intranuclear Ca2+ signals within individual nuclear lobes of neutrophils. AB - Cytosolic free Ca(2+)and intranuclear Ca(2+)behave similarly in human neutrophils. However, conventional laser scanning at 350 ms/frame resolution at lower than physiological temperatures demonstrates that (i) the nuclear fluo3 Ca(2+)signal persists longer than the cytosolic signal in some (but not all) nuclear lobes, (ii) the neutrophil nuclear membrane and fine inter-lobe filaments present barriers to diffusion of fluo3-Ca(2+)and lucifer yellow, and (iii) the diffusion barrier correlates with condensed chromosomal material on the nuclear envelope and blockage of the movement of fluo3-Ca(2+)into individual nuclear lobes. PMID- 12758085 TI - Ligand binding energy and catalytic efficiency from improved packing within receptors and enzymes. AB - Some small molecules bind to their receptors, and transition states to enzymes, so strongly as to defy current understanding. We show that in the binding of biotin to streptavidin, the streptavidin structure becomes better packed. We conclude that this contraction of the streptavidin structure promotes biotin binding. The improved packing is associated with positively cooperative binding, occurring with a benefit in enthalpy and a cost in entropy. Evidence indicating that catalytic efficiency can also originate via improved packing in some enzyme transition states, derived from the work of others, is presented. Negatively cooperative ligand binding is concluded to induce converse effects (less efficient packing, a cost in enthalpy, and a benefit in entropy). It applies to the binding of O(2) to haemoglobin, which indeed occurs with a hitherto unreported loosening of the amide backbones of the haemoglobin monomers. PMID- 12758087 TI - Immunocytochemical study of the glucocorticoid receptor in rat liver nuclei after hyperthermic stress. AB - The presence of glucocorticoid receptors (GR) in rat liver nuclei over a 24 h time period following hyperthermic stress at 41 degrees C was immunocytologically studied using unfixed nuclear smears. Liver nuclei in unstressed animals were found to be immunonegative for GR. However, intense GR immunopositivity followed by a subsequent gradual decrease in receptor levels was observed in the nuclei of test animals during the first 2 h after stress. This stress-related increase in the receptor nuclear level was greater than the increase seen after dexamethasone administration. These results suggest that hyperthermic stress could potentiate the hormonal stimulation of receptor nuclear translocation. PMID- 12758089 TI - Suppression of anoikis by v-Src but not by activated c-H-ras in human gallbladder epithelial cells. AB - Detachment of anchorage-dependent normal epithelial cells from their substratum causes the type of apoptosis known as anoikis, whereas malignant cells can proliferate independently of anchorage. Because src and ras oncogenes are activated in many human cancers, we investigated their role and downstream signaling pathways in anoikis resistance, using HAG-1 human epithelial cells transfected with v-src or activated H-ras. Consequently, anchorage-dependent mock or ras-transfected cells underwent anoikis. In contrast, anchorage-independent v Src-transformed cells did not exhibit such apoptotic features. Focal adhesion kinase (FAK), a transducer of integrin, was only activated in v-Src-transformed cells. Herbimycin A, an Src kinase inhibitor, reduced tyrosyl phosphorylation of FAK and reversed resistance to anoikis. However, both protein kinase C (PKC) and phophatidylinositol-3 (PI-3) kinase inhibitors failed to induce anoikis. These data suggest that the ability of activated Src to prevent anoikis may be mediated by Src to a downstream signaling pathway involving FAK, but not Ras, PI-3 kinase, or PKC. PMID- 12758088 TI - Growth control of mouse mammary epithelial cells by keratin antibody-targeted liposome containing oligonucleotides antisense to epidermal growth factor receptor. AB - NMuMG cells were incubated with 17beta-estradiol (E)+progesterone (P)+epidermal growth factor (EGF), with or without various types of oligomers (21-mers) to the EGF receptor activity domain (amino acid residues 718 to 724). Sense or antisense oligomers were encapsulated in protein A-bearing liposomes. Uncoupled protein A and unencapsulated sense or antisense oligomers were separated from liposomes on a Sepharose 4B column (the encapsulation efficiency of oligomers in liposome protein A was 0.8%). The addition of various concentrations of EGF to E+P showed an interaction between them during DNA synthesis (P<0.05). Antisense oligomers (1 microM) decreased DNA synthesis induced by E+P+EGF (65.0% inhibition, P<0.05). Sense oligomers also inhibited DNA synthesis induced by E+P+EGF (P<0.05). However, random-sequence oligomers did not inhibit EGF-induced DNA synthesis. We cannot rule out the possibility that sense oligomers match an unknown functional gene mRNA involved in cell growth, which causes their inhibitory effect. Cells were incubated with a keratin monoclonal antibody and then with dilutions of protein A-bound liposomes containing sense or antisense oligomers in the presence of E+P+EGF. Dose dependent inhibition of DNA synthesis was observed. The encapsulated oligomers in protein A-bound liposomes inhibited DNA synthesis at a 100-fold lower concentration than that of unencapsulated oligomers or the oligomer+liposome mixture. The tyrosine kinase activity domain has an important role in EGF regulation of mammary growth. The effect of a cytokeratin-targeted antibody on DNA synthesis in normal mouse mammary epithelial cells was marginal. PMID- 12758091 TI - Cytoskeletons in prokaryotes. AB - Not only eukaryotes, but also prokaryotes possess a cytoskeleton. Tubulin-related bacterial protein FtsZ, and actin-related bacterial proteins MreB/Mbl have recently been described as constituents of bacterial cytoskeletons. Genes coding for MreB/Mbl could only be found in elongated bacteria, not in coccoid forms. It was speculated that constituents of today's eukaryotic cytoskeleton (tubulin, actin) may have evolved from prokaryotic precursor proteins closely related to today's bacterial proteins FtsZ and MreB/Mbl. Prior to the description of proteins MreB/Mbl, evidence had been obtained for the existence of a shape preserving cytoskeleton ubiquitously present in all bacteria. In the meantime, structural studies allow to speculate on a possible role of bacterial elongation factor Tu (EF-Tu) as a structural element in such a "cytoskeletal web". EF-Tu was long known to form fibrillar structures in vitro; now experimental data accumulate, pointing towards formation of intracellular protofilaments containing EF-Tu, and networks thereof as well. In addition, results of these structural studies suggest a so far unknown mode of complex formation of EF-Tu with active ribosomes: ribosomes/polysomes were seen to be attached to intracellular protofilaments. Implications for the understanding of EF-Tu-ribosome interaction, and a role of such a kind of putative protofilaments as a general site of attachment for cellular functional macromolecules are discussed. The notion is discussed that an EF-Tu-containing cytoskeletal web might have been the "primary" or "basic" kind of prokaryotic cytoskeleton, already in existence prior to the "invention" of precursors of today's MreB. PMID- 12758090 TI - Effect of a single neonatal endorphin treatment on the hormone content of adult rat white blood cells and mast cells. AB - Using flow cytometry and confocal microscopy, the presence of endorphin, serotonin and chorionic gonadotropin (hCG) was demonstrated in rat white blood cells and peritoneal mast cells. After a single neonatal treatment with beta endorphin (hormonal imprinting), the mast cells of female rats reaching adulthood contained significantly less endorphin and serotonin, as well as slightly less hCG, than control cells. There was no change in the hormone content of the mast cells of males. The lymphocytes, monocytes and granulocytes of both sexes also contained the three hormones, but endorphin imprinting had no effect on these cells. PMID- 12758092 TI - Caution regarding the combined effects of extracellular matrices and nutrient media on cultured endothelial cells. AB - To study the influence of smooth muscle cells (SMC) on endothelial cells (EC), different co-culture designs are available, including EC seeding on SMC extracellular matrix (ECM). We explored human umbilical vein endothelial cell (HUVEC) adhesion and proliferation on either in situ or coated ECM, elaborated by HUVECs or human arterial smooth muscle cells (HUASMCs), in the presence of different nutrient media containing varying amounts of fetal calf serum. Coating wells with HUVEC or HUASMC ECMs did not improve HUVEC adhesion 1 h after cell seeding, compared with uncoated wells. HUVEC adhesion on in situ HUVEC-ECM and HUASMC-ECM was significantly increased compared with uncoated wells. The substratum upon which cells are maintained was found to play a crucial role, in conjunction with the medium to which HUVECs are exposed for their proliferative response. These results stress the importance of selecting media in relation to the particular substratum, in order to avoid misinterpretation of data. PMID- 12758093 TI - Unexpected durability of PKH 26 staining on rat adipocytes. AB - In a case of autologous mature fat cell transplantation to an individual rat, persistence of adipose staining with PKH26 was detected 14.5 months later. Fluorescent fat cells with spotty surface markings were easily visible. Such a long period of persistence and stability of staining exceeds the expectation from previous reports, and the given half-life of the substance. This finding encourages the use of the dye for long-term follow-up of connective tissue cells, especially adipocytes and preadipocytes, following transplantation. PMID- 12758094 TI - Subfertility: causes, treatment and outcome. AB - Common causes of subfertility include ovulatory disorders, tubal disease, peritoneal adhesions, endometriosis, uterine abnormalities, abnormalities of sperm and advancing female age. Infertility is unexplained after thorough evaluation in about 5-10% of cases. Significant caveats must be attached to the interpretation of available data regarding infertility treatments. Successful ovulation induction in anovulatory women is possible for nearly all women except in cases of ovarian failure. Surgery is an option for some patients with tubal damage, adhesions, endometriosis and uterine abnormalities. Male factor infertility may be amenable to treatment of a specific cause, but is often empirical with the use of intra-uterine insemination (IUI) or in vitro fertilization (IVF). Egg donation is currently the most effective treatment available for age-related infertility when other treatments have not been successful. Couples with unexplained infertility may be effectively treated with ovulation induction plus IUI or IVF. PMID- 12758095 TI - Advances in the assessment of the uterus and fallopian tube function. AB - Hysterosalpingo-contrast-sonography (HyCoSy) using saline and Echovist is a well tolerated outpatient technique that provides a significant amount of information of relevance to the infertile woman that is not obtainable at hysterosalpingogram (HSG) whilst avoiding exposure to X-ray irradiation. When performed by experienced operators, it serves as a valuable, first-line screening test for the more invasive procedures of laparoscopy and dye chromopertubation and hysteroscopy. If detailed diagnostic information is required in women in whom there is no clinical or ultrasound evidence of pelvic pathology, the surgical technique of fertiloscopy can be considered to be appropriate. This technique permits confirmation that the ovum pick-up mechanism is normal, the tubes are patent and the uterine cavity is normal, while salpingoscopy and microsalpingoscopy permit the assessment of the tubal lumen. PMID- 12758096 TI - Male infertility. AB - Infertility affects 13-18% of couples and growing evidence from clinical and epidemiological studies suggests an increasing incidence of male reproductive problems. The pathogenesis of male infertility can be reflected by defective spermatogenesis due to pituitary disorders, testicular cancer, germ cell aplasia, varicocele and environmental factors or to defective sperm transport due to congenital abnormalities or immunological and neurogenic factors. Recent studies suggest an increased incidence of genetic disorders related to male infertility which may affect different levels, interfering with germ cell generation and maturation or leading to the production of non-functional spermatozoa. The identification of genetic causes of male infertility raises the issue of the transmission of defects to the offspring, a situation that is becoming more important given the increasing use of intracytoplasmic sperm injection (ICSI), a procedure in which the natural selection of the spermatozoa is by-passed. Fertilization can occur in vitro using ejaculated, epididymal or testicular spermatozoa, either fresh or frozen-thawed, providing opportunities hitherto not possible for men to be genetic fathers. PMID- 12758097 TI - Developments in drugs for ovarian stimulation. AB - Drugs for ovarian stimulation have been improved during the last decades. Initially gonadotrophins were extracted from human pituitary glands and urine; nowadays they are produced from transformed cell-lines. All three gonadotrophins- follicle stimulating hormone (FSH), luteinizing hormone (LH) and human chorionic gonadotrophin (hCG)--are now marketed as recombinant (r-) products. The near-100% pure FSH preparations might, in some situations, cause abnormally low LH levels and it is likely that the addition of LH may be beneficial in these situations. It is possible that r-LH will become available in sufficient dosages to replace hCG for ovulation induction and this may reduce the incidence of ovarian hyperstimulation syndrome due to its shorter half-life. In parallel to the development of gonadotrophin preparations, protocols for ovarian stimulation are now more comfortable for the patients, especially with the introduction of gonadotrophin receptor hormone (GnRH)-agonists in the early 1980s and, more recently, the introduction of GnRH-antagonists. PMID- 12758098 TI - Prevention and treatment of ovarian hyperstimulation syndrome. AB - The ovarian hyperstimulation syndrome (OHSS) is a potentially fatal condition with a pathophysiology that is not clearly understood. A shift in fluid from the extravascular space occurs, thought to be induced by cytokines and/or vascular endothelial growth factor. Human chorionic gonadotrophin (hCG), exogenous or endogenous, seems to be the triggering mechanism, resulting in early and late development of the syndrome, respectively. The management of the syndrome is mainly symptomatic. Preventive strategies are being developed and constantly refined. Women at increased risk of OHSS need to be on the lowest possible dose of gonadotrophin with the aim of reducing the granulosa/luteal cell mass. Ultrasound and serum oestradiol (E2) measurements are, at present, the main methods used to identify and monitor those at risk during controlled ovarian hyperstimulation (COH). Withholding gonadotrophin stimulation (coasting), but continuing down-regulation, when a large number of follicles (greater than 20) and a rising serum oestradiol level are seen, is the most widely favoured and used preventive measure and the most cost effective. Management is symptomatic and aimed at achieving fluid balance, restoring plasma volume and improving renal function. This may be combined with an early resort to ascitic fluid aspiration, which will improve the feeling of wellbeing and may remove those agents responsible for the syndrome. Heparin, to prevent the risk of thromboembolism as a result of haemoconcentration, is important. PMID- 12758099 TI - Polycystic ovary syndrome--a systemic disorder? AB - Polycystic ovary syndrome (PCOS) is the commonest endocrine disturbance affecting women. There is considerable heterogeneity of symptoms and signs amongst women with PCOS and for an individual these may change over time. PCOS is familial and genetics appear to be associated with disturbances of insulin secretion. Polycystic ovaries can exist without clinical signs of the syndrome, which may then become expressed over time. There are a number of interlinking factors that affect expression of PCOS. A gain in weight is associated with a worsening of symptoms whilst weight loss will ameliorate the endocrine and metabolic profile and symptomatology. Women with PCOS are characterized by the presence of insulin resistance, central obesity and dyslipidaemia, which appears to place them at a higher risk of developing diabetes as well as cardiovascular disease. A number of studies have confirmed the higher incidence of diabetes, although they have not shown a higher risk of mortality from ischaemic heart disease (IHD). Cross sectional studies have demonstrated a significant association between PCOS and IHD. Prospective, long-term longitudinal studies confirming this risk are still awaited. Weight loss remains the most effective first line therapeutic intervention in women with PCOS and recently the use of insulin sensitizing agents has demonstrated benefit. PMID- 12758100 TI - Endometriosis: modern surgical management to improve fertility. AB - Endometriosis is a pleiotropic reproductive condition and the lesions visualized at laparoscopy are only one aspect of this disease process. At present, there is no evidence that surgery for endometriosis can cure infertility. The most important surgery in infertility is ovarian surgery. It is generally accepted that, in most cases, the invagination of the cortex results in the formation of an endometriotic pseudocyst. As a consequence, primordial follicles are present at the base of the cyst. Since surgery is of limited value, the utmost care has to be given to conservation of the patient's fertility. Ablative surgery offers advantages over excision of the cyst in terms of less adhesion formation and better preservation of the ovarian reserve. The ablative eversion technique differs from fenestration and drainage by its access through the site of inversion and resection of the fibrotic ring. Using the new technique of transvaginal hydrolaparoscopy, access to the site of pathology in the fossa ovarica is facilitated. Furthermore, the aqueous distension medium keeps the organs afloat and provides a clear delineation between the organs and adhesions allowing atraumatic reconstructive surgery. PMID- 12758101 TI - The endometrium as a cause of implantation failure. AB - One of the biggest obstacles to reproductive success is our inability to diagnose or treat effectively the non-receptive endometrium. The endometrium becomes receptive for a limited period of time under the influence of steroid hormones and paracrine signals from the developing embryo. It is likely that the receptive state is characterized by the expression of particular genes that allow the normally refractory endometrium to respond to the embryo and permit attachment. Recently, several molecules have been reported whose function is essential for uterine receptivity in rodents and primates. This article will review current models of the control of receptivity and early implantation and assess progress in defining markers for the receptive endometrium in women. Although some genes have been identified whose expression is altered in women with subfertility, none of these molecular markers have yet proven clinically useful in the assessment of functional receptivity. The use of high throughput techniques such as microarrays and proteomic methods to investigate gene expression in the endometrium provides a novel approach to defining receptivity at the molecular level. The potential impact of these tools on clinical practice will be discussed. PMID- 12758102 TI - Reducing the incidence of twins and triplets. AB - Multiple pregnancy rates remain high after assisted conception because of a misconceived assumption that transferring three or more embryos will maximize pregnancy rates. Maternal morbidity is sevenfold greater in multiple pregnancies than in singletons, perinatal mortality rates are fourfold higher for twins and sixfold higher for triplets, while cerebral palsy rates are 1-1.5% in twin and 7 8% in triplet pregnancies. Therefore, multiple pregnancies must be considered a serious adverse outcome of assisted reproductive techniques. Primary prevention of multiple pregnancies is the solution. The overwhelming evidence presented in this chapter demonstrates that limiting the embryo transfer in in vitro fertilization to two embryos would significantly reduce adverse maternal and perinatal outcomes by reducing the incidence of high order multiple pregnancies without reducing take-home-baby rates. Secondary prevention by multifetal pregnancy reduction is effective, but not acceptable to all patients. New developments in blastocyst culture, single embryo transfer, embryo cryopreservation and pre-implantation aneuploidy exclusion, should allow improvements in pregnancy rates without increasing multiple pregnancies. PMID- 12758103 TI - Cryobiology of ovarian and testicular tissue. AB - Cryopreservation of ovarian and testicular tissue can be used in young individuals who are likely to lose their gonadal function prematurely because of chemotherapy, radiotherapy or a known genetic condition causing gonadal failure. Tissue cryopreservation is relatively well established, but freezing techniques have still to be optimized. Slow programmed freezing using propanediol or dimethylsulphoxide as a cryoprotectant is the most established method. Tissue cryopreservation is highly experimental. No human pregnancies have been achieved as yet, although re-implantation experiments have been started. Maturation of gametes in vitro from cryopreserved tissue is far from being clinically applicable. Cryopreservation of spermatozoa and embryos is an established method, whenever applicable. Cryopreservation of oocytes is still experimental, but successful pregnancies have been achieved by several clinics. Cryopreservation of testicular spermatozoa for intracytoplasmic sperm injection (ICSI) is an established method and should be performed whenever a testicular biopsy is being carried out. PMID- 12758104 TI - Cancer risks associated with the diagnosis of infertility. AB - The effects of infertility and its treatment on cancer risk are of concern to many infertile couples. Infertile women appear to be at no higher risk for breast cancer although they may be more at risk for cancers of the uterus, particularly if they have ovulation disorders. Most studies show no increase in the risk of invasive ovarian cancer in infertile women, but ovarian tumours of borderline malignancy are more common. There seems to be no overall increase in cancer risk associated with fertility drug treatment, but questions remain about risk in subgroups of infertile women. Male infertility has been associated with an increased risk of testis cancer. Cancer incidence in children born after fertility treatment appears to be similar to that in the general population. Discussion of cancer risks in the context of other reproductive and community risks can be helpful to patients. PMID- 12758108 TI - Impact of aging on hippocampal function: plasticity, network dynamics, and cognition. AB - Aging is associated with specific impairments of learning and memory, some of which are similar to those caused by hippocampal damage. Studies of the effects of aging on hippocampal anatomy, physiology, plasticity, and network dynamics may lead to a better understanding of age-related cognitive deficits. Anatomical and electrophysiological studies indicate that the hippocampus of the aged rat sustains a loss of synapses in the dentate gyrus, a loss of functional synapses in area CA1, a decrease in the NMDA-receptor-mediated response at perforant path synapses onto dentate gyrus granule cells, and an alteration of Ca(2+) regulation in area CA1. These changes may contribute to the observed age-related impairments of synaptic plasticity, which include deficits in the induction and maintenance of long-term potentiation (LTP) and lower thresholds for depotentiation and long term depression (LTD). This shift in the balance of LTP and LTD could, in turn, impair the encoding of memories and enhance the erasure of memories, and therefore contribute to cognitive deficits experienced by many aged mammals. Altered synaptic plasticity may also change the dynamic interactions among cells in hippocampal networks, causing deficits in the storage and retrieval of information about the spatial organization of the environment. Further studies of the aged hippocampus will not only lead to treatments for age-related cognitive impairments, but may also clarify the mechanisms of learning in adult mammals. PMID- 12758109 TI - Estrogens: protective or risk factors in brain function? AB - Over the past century, the average lifespan of women has increased from 50 to over 80 years, but the age of the menopause has remained fixed at 51 years. This "change of life" is marked by a dramatic and permanent decrease in circulating levels of ovarian estrogens. Therefore, more women will live a greater proportion of their lives in a chronic hypoestrogenic state. Ovarian steroid hormones are pleiotropic and have multiple, diverse, and possibly opposing actions in different contexts. In light of recent reports of the possible health risks of hormone replacement therapy (HRT) on several different physiological systems, the question of whether estrogens are protective or risk factors must be carefully re evaluated. PMID- 12758110 TI - Proteomics in brain research: potentials and limitations. AB - The advent of proteomics techniques has been enthusiastically accepted in most areas of biology and medicine. In neuroscience, a host of applications was proposed ranging from neurotoxicology, neurometabolism, determination of the proteome of the individual brain areas in health and disease, to name a few. Only recently, the limitations of the method have been shown, hampering the rapid spreading of the technology, which in principle consists of two-dimensional gel electrophoresis with in-gel protein digestion of protein spots and identification by mass-spectrometrical approaches or microsequencing. The identification, including quantification using specific software, of brain protein classes, like enzymes, cytoskeleton proteins, heat shock proteins/chaperones, proteins of the transcription and translation machinery, synaptosomal proteins, antioxidant proteins, is a clear domain of proteomics. Furthermore, the concomitant detection of several hundred proteins on a gel allows the demonstration of an expressional pattern, rather generated by a reliable, protein-chemical method than by immunoreactivity, proposed by protein-arrays. An additional advantage is that hitherto unknown proteins, so far only proposed from their nucleic acid structure, designated as hypothetical proteins, can be identified as brain proteins. As to shortcomings and disadvantages of the method we would point to the major problem, the failure to separate hydrophobic proteins. There is so far no way to analyse the vast majority of these proteins in gels. Several other analytical problems need to be overcome, but once the latter problem can be solved, there is nothing to stop the method for a large scale analysis of membrane proteins in neuroscience. PMID- 12758111 TI - Neurovascular diseases: from angiography to management. PMID- 12758112 TI - Treatment of ruptured intracranial aneurysms: implications of the ISAT on clipping versus coiling. AB - Patients with a ruptured intracranial aneurysm should be treated as soon as possible after the haemorrhage to prevent rebleeding and to allow vigorous treatment of ischemic events in case of vasospasm. The choice of treatment, endovascular or surgical, should be based on the angio-architectural aspects of the aneurysm. 3D rotational subtraction angiography will more clearly show the aneurysm morphology and will therefore help in the decision-making process. If an aneurysm is suitable for endovascular treatment ('coiling'), this should be the treatment of first choice, as has been clarified in the ISAT study. Location of the aneurysm only influences the treatment decision in aneurysms located at the basilar artery bifurcation. These aneurysms are preferably treated by endovascular means. The long-term results of endovascular and surgical treatment are still the subject of debate. For both treatment modalities, re-growth of the treated aneurysm has been described, but solid comparative data is missing. In analogy with the ISAT, referral of patients with a ruptured intracranial aneurysm should be performed as soon as possible after the haemorrhage. Preferably, this should be a hospital where neurosurgeons, interventional neuroradiologists, as well as neurologists (with expertise on medical treatment of patients with a subarachnoid haemorrhage) collaborate. PMID- 12758113 TI - Giant intracranial aneurysms: development, clinical presentation and treatment. AB - The natural history of giant intracranial aneurysms are grave. More than 50% of patients suffer from rupture of these aneurysms and mortality is >60% in 2 years. Modern technology and advancement of knowledge in neurosurgery and interventional neuroradiology have altered its natural course for the better. As many reports have shown, the majority of these aneurysms can be treated either by surgery or by endovascular approach, even though morbidity is higher than when treating smaller aneurysms. Certain aneurysms are more suitable to direct surgical clipping and others may have better chances of good clinical outcome by endovascular treatment. It is imperative to analyse the location, morphology, hemodynamics and circulation of normal brain of each aneurysm before the mode of treatment is decided. Needless to say, the individual patient's age, neurological and medical condition should be considered. For endovascular treatment, application of each technique, endosaccular occlusion or parent artery occlusion depends on the aneurysm location and geometry as well as its pathology. Several reports indicated that clinical outcome is better in patients treated by parent artery occlusion since it eliminates any blood flow to the aneurysm and it provides a more effective reduction of the mass effect. However, not all parent arteries can be sacrificed. In addition, endosaccular treatment is effective in preventing haemorrhage if the aneurysm is not re-canalised. It is also demonstrated that symptoms of mass effect can be reversed by endosaccular coiling. The patients who are treated this way should be closely monitored for re canalisation. PMID- 12758114 TI - Management of patients with brain arteriovenous malformations. AB - Arteriovenous malformations (AVMs) of the brain, which are probably genetically determined, are errors in the development of the vasculature that, together with the effects of blood flow, may lead to a focal arteriovenous shunt. Clinically, the adult patient may present with acute or chronic neurological symptoms-fixed or unstable-such as deficits, seizures or headache. Sometimes the lesion is an incidental finding. In about half of the patients, the revealing event is an intracranial haemorrhage. The prevalence of AVM in the western world is probably <0.01% and the detection rate is about one per 100,000 person-years. Most AVMs are revealed in patients 20-40 years of age. Therefore, the risk of developing neurological symptoms from an AVM, usually because of haemorrhage, increases with patient age. In the young adult population, AVMs are significant risk factors for hemorrhagic stroke. This risk increases with AVM volume and is higher in centrally located AVMs. Almost all patients with AVM are subjected to treatment, either by surgery, radiosurgery or embolisation, with the functional aim of reducing the risk of haemorrhage or to alleviate neurological symptoms with an acceptable treatment risk. Few neurocentres have physicians highly skilled in all treatment modalities. Therefore, the prescribed treatment may not be defined from an objective assessment of what is optimal for each individual patient, but rather from local expertise. In this context, more and better data about the natural history and the outcome of different treatments, as well as predictive models, would be valuable to help to optimise the management. Management strategies obviously differ according to local preferences, but results presented in the literature suggest the following strategy: (I) cortically located AVMs with a nidus volume <10 ml could be operated, with or without presurgical embolisation, unless there is a single feeder that can easily be catheterised and embolised for obliteration or other obvious target for embolisation, such as pseudoaneurysms or large fistulae; (II) centrally located AVMs with a nidus volume <10 ml should be treated by radiosurgery, unless suitable for embolisation as indicated above; (III) patients harbouring AVMs with a nidus volume >10 ml could benefit from targeted partial embolisation followed by radiosurgery or surgery, depending on the angioarchitecture; and (IV) AVMs >20 ml nidus volume usually have a high treatment risk with any treatment modality and are not obvious targets for treatment at all. PMID- 12758115 TI - Management of intracranial dural arteriovenous shunts in adults. AB - Dural arteriovenous shunts are abnormal arteriovenous communications within the dura. They are thought to be an acquired condition in adults and can present with a variety of clinical features, ranging from benign bruits to intracranial hemorrhage and neurological deficits. The presentation and natural history of these shunts is largely determined by the pattern of venous drainage. Knowledge of natural history and careful study of the angioarchitexture by angiography is therefore mandatory for correct management of these lesions. In this review, principles of management in adults and the various factors that influence treatment decisions are discussed, with a focus on endovascular therapy. Retrograde leptomeningeal or cortical venous drainage has a strong correlation with adverse clinical events and the requirement for aggressive management in this situation is highlighted. Indications for endovascular treatment, therapeutic goals, approaches and techniques are reviewed. The role of surgical treatment is also briefly discussed. PMID- 12758117 TI - Primary thyroid lymphoma: CT findings. AB - INTRODUCTION: To evaluate the computed tomographic (CT) findings of primary thyroid lymphoma. METHODS AND MATERIAL: The clinicopathological data and CT images of nine patients with primary thyroid lymphoma were retrospectively reviewed. The CT appearances were classified into three types: type 1, a solitary nodule surrounded by normal thyroid tissue; type 2, multiple nodules in the thyroid, and type 3, a homogeneously enlarged both thyroid glands with a reduced attenuation with or without peripheral thin hyperattenuating thyroid tissue. RESULTS: All patients had a rapidly enlarging thyroid mass and coexistent Hashimoto's thyroiditis. One patient showed type 1 pattern, three type 2, and five type 3. Six patients had homogeneous tumor isoattenuating to surrounding muscles. The tumors had a strong tendency to compress normal remnant thyroid and the surrounding structure without invasion. CONCLUSION: Primary thyroid lymphoma should be included in the differential diagnosis when old female had a homogeneous thyroidal mass isoattenuating to muscles, which does not invade surrounding structures. PMID- 12758116 TI - Spinal cord arteriovenous shunts: from imaging to management. AB - Spinal cord arteriovenous shunts (SCAVSs) are either fistulas or niduses that can be separated in four different groups according to their localization and relationship to the dura. Paraspinal AVSs are located outside the spine and are responsible for neurological symptoms because of cord compression by ertatic veins, venous congestion or arterial steal. Epidural shunts are located in the epidural space and drain in epidural veins with secondary intradural congestion. Dural shunts are embedded in the dura, produce a cord venous myelopathy after draining through veins that either pierce the dura far from a nerve root or accompany a nerve root. Intradural shunts affect the cord, the roots or the filum. Additionally, they can be classified according to their potential relationships with genetics, vascular biological features and angiogenesis into genetic hereditary lesions (hereditary hemorrhagic telangiectasia), genetic non hereditary lesions (multiple lesions with metameric links) and single lesions (AVMs or micro AVFs). MRI and MRA are able to visualise SCAVS early after the onset of clinical symptoms. The type of shunt and its localization may remain difficult to be precise. Angiography remains the gold standard for analysis of the anatomical, morphological and architectural features necessary for therapeutic decisions in both paediatric and adult populations. In our series, embolisation is chosen in first intention whatever the type of shunt responsible for the clinical symptoms and glue is preferably used. In paraspinal, dural or epidural arteriovenous shunts, the goal of treatment should be complete closure of the shunt. A complete cure by embolization is rather easily achieved in paraspinal lesions. Failure of endovascular therapy in dural or epidural shunts must bring the patient to surgery. The prognosis of most intradural shunts seems better than previously thought, even after haemorrhage. In intradural spinal cord arteriovenous shunts, embolisation targeted towards the portions of the malformation felt to be responsible for the symptoms (venous congestion) or pointing to the point of rupture (false aneurysms) of the malformation, allows restoration of a new hemodynamic equilibrium between the malformation and the cord itself. Such targeted treatment offers long-term stabilisation or improvement to patients suffering from SCAVSs and good protection against (re) haemorrhages, with an acceptable morbidity. Cure of the shunt is not imperative to obtain these satisfactory outcomes. The clinical results obtained by such management compare favourably with those obtained by neurosurgery. PMID- 12758118 TI - Physicians' liability in interventional radiology and endovascular therapy. AB - INTRODUCTION/OBJECTIVE: Modern practice in Radiology has rapidly changed over the last decades incorporating invasive techniques. Additionally, litigation in medical practice has arisen as an important issue. This article aims to highlight issues related to malpractice in interventional radiology and endovascular therapy in order to point out the importance of the written informed consent. METHODS AND MATERIAL: Search of relevant literature from the Pubmed. RESULTS: The role of radiologist has been greatly transformed over the last decades. He is not only entitled to participate in the diagnosis but also he undertakes therapeutic procedures, either alone or as a member of a team. Thus the radiologist is now more exposed to actions that maximize litigation risk. Adequate communication and a written consent form seem to be mandatory before any invasive radiological procedure. Patient should know in detail the benefits and the risks of the scheduled procedure and whether the proposed therapy is a new form of treatment or part of a randomized trial. DISCUSSIONS AND CONCLUSION: Interventional radiologist or physician is exposed to high litigation risk. This certainly requires an urgent adaptation of his practice and attitude to the new reality. Written patient's informed consent remains an integral part of the communication between physicians and patients, and importantly is offering professional protection along these lines. PMID- 12758119 TI - Ophthalmic artery color Doppler ultrasonography in mild-to-moderate preeclampsia. AB - OBJECTIVE: To evaluate the hemodynamic changes in mild-moderate preeclampsia using ophthalmic artery Doppler ultrasonography technique. METHODS: Ophthalmic artery pulsatility and resistivity indices were calculated in 30 mild-moderate preeclamptic women and 30 normotensive gravid women of matched gestational age. Student's t-test was performed to test the significance of difference. RESULTS: Both indices of peripheral resistance were found to be significantly lower in the ophthalmic arteries of mild-moderately preeclamptic women than those measured in normotensive gravid women at similar stage of pregnancy. In a small number patients whose disease progressed to severe preeclampsia, both indices increased. CONCLUSION: In patients with mild-moderate preeclampsia, ophthalmic artery color Doppler ultrasonography detects hemodynamic changes that are not present in normotensive gravid women. Reversal of Doppler patterns in a small number of patients with progressive disease supports the hypotheses suggesting the presence of early vasodilation and late vasospasm in the etiology of preeclampsia. PMID- 12758122 TI - Identification of conserved vault RNA expression elements and a non-expressed mouse vault RNA gene. AB - Vault RNA (vRNA) genes have been cloned from several vertebrates including rat, mouse, and humans. Their copy numbers vary, as does the length of the encoded RNA. We have determined that the mouse genome contains two vRNA genes; one is expressed the other is a pseudogene. In vitro transcription of the rat vRNA gene by RNA polymerase III has previously been shown to be dependent on a combination of both external and internal promoter sequence elements. By comparing the upstream regions of the vertebrate vRNA genes, a 25 bp conserved sequence and a TATA box can be identified. Furthermore, the unique arrangement of the internal promoter boxes (one A and two B boxes) is conserved in the expressed human vRNA genes even though a new RNA polymerase III termination sequence has evolved between the two B boxes. PMID- 12758123 TI - Unusual number and genomic organization of Hox genes in the tunicate Ciona intestinalis. AB - Hox genes are organized in genomic clusters. In all organisms where their role has been studied, Hox genes determine developmental fate along the antero posterior axis. Hence, these genes represent an ideal system for the understanding of relationships between the number and expression of genes and body organization. We report in this paper that the ascidian Ciona intestinalis genome appears to contain a single Hox gene complex which shows absence of some of the members found in all chordates investigated up to now. Furthermore, the complex appears to be either unusually long or split in different subunits. We speculate that such an arrangement of Hox genes does not correspond to the chordate primordial cluster but occurred independently in the ascidian lineage. PMID- 12758124 TI - Plac8 and Plac9, novel placental-enriched genes identified through microarray analysis. AB - Microarray expression profiling of a collection of 15,000 mouse genes with placental and embryonic RNAs revealed candidates for placental-enriched genes, three of which we have confirmed and further characterized. One, Plac1, strongly expressed in all trophoblast-derived cells in the placenta, has been described earlier (Genomics 68 (2000) 305). Here we report that of the other two, Plac8 expression is restricted to the spongiotrophoblast layer during development, whereas Plac9 is weakly expressed though highly enriched in placenta. For both, cDNAs with complete open reading frames were recovered and exon-intron structures inferred from comparisons of mouse cDNA and genomic sequence. The predicted proteins (112 and 108 amino acids) both contain putative signal peptides, with a coiled-coil segment of mPLAC9 as the only other detected motif. Genomic sequence comparisons reveal that in addition to an apparent pseudogene on chromosome 1, Plac8 is expressed at mouse cytoband 5e3. It is tightly conserved in human in a syntenically equivalent ortholog at 4q21.23. Plac9 is present in a single copy on chromosome 14, with a syntenically equivalent human ortholog at 10q22.3. Putative promoter regions up to 10 kb 5' of the transcription units for Plac1, Plac8, and Plac9 contain sites for widely-expressed transcription factors which, by analogy to other instances, may be sufficient to explain placental enrichment. PMID- 12758125 TI - Identification and cloning of two forms of liver peroxisomal fatty Acyl CoA Oxidase from the koala (Phascolarctos cinereus). AB - In the present study, the cloning, expression and characterization of the rate limiting enzyme of the peroxisomal beta-oxidation spiral, acyl CoA oxidase (AOX), from koala (Phascolarctos cinereus) liver is described. It has been previously reported that peroxisomal cyanide-insensitive palmitoyl-CoA oxidation activity was absent in koala liver [Comp. Biochem. Physiol. (C) 127 (2000) 327]. This activity is a measure of the overall peroxisomal beta-oxidation minus the final step catalysed by thiolase. Two 2039 bp koala liver AOX cDNAs, designated AOX1 and AOX2, were cloned by reverse transcription-polymerase chain reaction and rapid amplification of cDNA ends. The koala AOX cDNAs encode proteins of 662 amino acids. Transfection of the koala AOX cDNAs into Cos-7 cells resulted in the expression of proteins with palmitoyl-CoA oxidase activity. The apparent K(m) values for AOX1 and AOX2 cDNA-expressed enzymes were 28 and 38 microM, respectively, which are within the range of order of magnitude reported for rat and human purified AOX enzymes (approximately 10 microM). Northern analysis, utilizing the koala AOX1 cDNA as probe, detected a more intense AOX mRNA band in the koala liver as compared to rats and humans. Southern blot analysis of liver genomic DNA samples revealed a single AOX gene fragment of less than 14 kb in koalas, rat and humans, suggesting a single AOX gene. Collectively, the results of this study suggest that the absence of peroxisomal cyanide-insensitive palmitoyl-CoA oxidation activity in the koala liver is possibly due to deficiencies of one or more enzymes downstream of acyl-CoA oxidase and/or deficiencies of mitochondrial beta-oxidation enzymes. PMID- 12758126 TI - Transcription control of a gene for Drosophila transcription factor, DREF by DRE and cis-elements conserved between Drosophila melanogaster and virilis. AB - A DNA replication-related element (DRE)-binding factor (DREF) has been revealed to be an important transcription factor for activating promoters of cell proliferation and differentiation related genes. The amino acid sequences of DREF are conserved in evolutionary separate Drosophila species, Drosophila melanogaster (Dm) and Drosophila virilis (Dv) in three regions. In the present study, evidence was obtained that there are several highly conserved regions in the 5' flanking region between the DmDREF and DvDREF genes. Band mobility shift assays using oligonucleotides corresponding to these conserved regions revealed that specific trans-acting factors can bind to at least three regions -554 to 543 (5'-TTTGTTCTTGCG), -81 to -70 (5'-GCCCACGTGGCT) and +225 to +234 (5' GCAATCAGTG). Using a transient luciferase expression assay, we demonstrated that the region -554 to -543 functions as a negative regulatory element for DmDREF promoter activity, while the regions -77 to -70 (5'-ACGTGGCT) and +225 to +236 (5'-GCAATCAGTGTT) function as positive regulatory elements. In previous studies, we observed that expression of the homeodomain protein Zerknullt (Zen) represses PCNA gene transcription, by reducing the DNA binding activity of DREF. Here we show Zen downregulates DREF gene promoter activity through action on the region between +241 and +254 (5'-AGAATACTCAACA). In addition, the DmDREF promoter contains five DREs. Using a double stranded RNA-mediated interference method, we generated evidence that expression of DmDREF could be auto-regulated by DREF through the third DRE located at +211 to +218. In living flies we obtained results consistent with those obtained in vitro and in cultured cells. The study thus indicates that DmDREF is effectively regulated via highly conserved regions between the DmDREF and DvDREF promoters, suggesting the existence of common regulatory factors, and that DmDREF can be positively regulated by itself via the third DRE located in its most highly conserved region. PMID- 12758127 TI - Characterization of the promoter and the transcription factors for the mouse UDP Gal:betaGlcNAc beta1,3-galactosyltransferase gene. AB - Galbeta1-3Gal-NAcbeta1-4Gal(3-2alphaNeuAc)beta1-4Glcbeta1-1Cer (GM1) is one of the most extensively investigated gangliosides that plays critical roles in the development and functions of the nervous system. UDP-Gal:betaGlcNAc beta1,3 galactosyltransferase (Gal-T-II) is responsible for synthesis of ganglioside GM1 in the ganglioside biosynthetic pathway. To understand the transcriptional regulation of Gal-T-II gene expression, we cloned a 1448 bp 5'-flanking fragment from the mouse Gal-T-II gene. The transcriptional activity of the fragment was demonstrated in mouse Neuro-2a cells by a luciferase assay. The proximal 550 bp fragment showed the highest transcriptional activity as determined by a series of 5'-truncated constructs of the promoter. One negative regulatory region was also identified. Primer extension assay revealed a transcription initiation site approximately 242 bp upstream from the ATG translation start codon. Analysis of the promoter sequence revealed a number of potential binding sites for known transcription factors. To determine which transcription factors bind to the promoter, we carried out a systematic search for the binding proteins using the 1142 bp Gal-T-II promoter fragment containing both positive and negative regulatory regions in a combination of DNA pull-down assay and transcription factor array analysis. Twenty-seven transcription factors bound to consensus sites in the promoter region. In addition, four other factors without consensus binding sites in this region were also recruited, possibly as components of transcription factor complexes. These data indicate that the basal regulation of Gal-T-II gene transcription involves multiple transcription factors, some of which may be present in complexes. PMID- 12758129 TI - Analysis of the 3' Cmu region of the rabbit Ig heavy chain locus. AB - The immunoglobulin D (IgD) antibody class was, for many years, identified only in primates, rodents and teleost fish. The limited distribution of IgD among vertebrates suggested that IgD is a functionally redundant antibody class that has been lost by many vertebrate species during evolution. The recent identification of IgD in artiodactyls, however, suggests that IgD might be more widely expressed among vertebrates than previously thought, possibly serving a unique role in immunity. IgD expression has been searched for but not detected in rabbits. In order to search directly for a rabbit Cdelta locus encoding the constant region of IgD, we determined the nucleotide sequence of 13.5 kb of genomic DNA downstream of the rabbit Cmu locus. We did not find a rabbit Cdelta locus in this region, but found instead that this region is densely populated by repetitive elements, including a long interspersed DNA element repeat, six C repeats, and two processed pseudogenes. We conclude that the rabbit probably does not express IgD because there is no Cdelta locus immediately downstream of the rabbit Cmu locus. PMID- 12758128 TI - The human SPANX multigene family: genomic organization, alignment and expression in male germ cells and tumor cell lines. AB - Multigenicity is one of the features of cancer/testis-associated genes. In the present study we analyzed the number and expression of genes of the SPANX(CTp11) family of cancer/testis-associated genes. Genomic database analysis, next to the four previously described SPANX genes, revealed the presence of a novel gene: SPANXE. Moreover, we detected an allelic variant of SPANXB resulting in one amino acid substitution in the encoded protein: SPANXB'. Most SPANX genes are present on contig NT_011574 located at Xq26.3-Xq27.1. Based on expressed sequence tag databases and RT-PCR analysis three additional novel SPANX sequences were identified, though not represented so far in the human genome sequence. Sequence alignments justify a subdivision of this gene family based on the absence (SPANXA likes) or presence (SPANXB) of an 18 base pair sequence stretch in the open reading frame. The alignments also reveal an unusually high level (99%) of intron homology. Furthermore, the nucleotide variations in the open reading frame almost all lead to amino acid substitutions. Southern blot and database analyses indicate that SPANX sequences are exclusively present in primates. With RT-PCR analysis on human sperm cell precursors and tumor cell lines most family members could be detected. SPANXB was only found in sperm cell precursors and could not be detected in the tumor cell lines tested. Overall SPANXA was the most frequently expressed SPANX variant in melanoma and glioblastoma cell lines. PMID- 12758130 TI - Regulated gene expression from adenovirus vectors: a systematic comparison of various inducible systems. AB - Positively and tightly regulated gene expression is essential for gene function and gene therapy research. The currently-used inducible gene expression systems include tetracycline (Tet-on and T-REx), ecdysone, antiprogestin and dimerizer based systems. Adenovirus (Ad) vectors play an important role in gene function and gene therapy research for their various advantages over other vector systems. Previously, we reported the inferiority of the Tet-on system as an inducible gene expression system in the context of Ad vectors in comparison with the Tet-off system. In this study, to identify an optimal system for regulated gene expression from Ad vectors, we made a rigorous direct comparison of these five inducible gene expression systems in three cell lines using the luciferase reporter gene. The highest sensitivity to the respective inducer was that of the dimerizer system, followed by the antiprogestin system. The lowest basal expression and the highest induction factor were both characteristic of the dimerizer system. Furthermore, the dimerizer and T-REx systems exhibited much higher induced expression levels than the other three systems. The elucidation of the characteristic features of each system should provide important information for widespread and feasible application of these systems. Overall, these results suggest the most appropriate inducible gene expression system in the context of Ad vectors to be the dimerizer system. PMID- 12758131 TI - Segmentation of volumetric tissue images using constrained active contour models. AB - In this article we describe an application of active contour model for the segmentation of 3D histo-pathological images. The 3D images of a thick tissue specimen are obtained as a stack of optical sections using confocal laser beam scanning microscope (CLSM). We have applied noise reduction and feature enhancement methods so that a smooth and slowly varying potential surface is obtained for proper convergence. To increase the capture range of the potential surface, we use a combination of distance potential and the diffused gradient potential as external forces. It has been shown that the region-based information obtained from low-level segmentation can be applied to reduce the adverse influence of the neighbouring nucleus having a strong boundary feature. We have also shown that, by increasing the axial resolution of the image stack, we can automatically propagate the optimum active contour of one image slice to its neighbouring image slices as an appropriate initial model. Results on images of prostate tissue section are presented. PMID- 12758132 TI - Compression assessment based on medical image quality concepts using computer generated test images. AB - Compression algorithms are widely used in medical imaging systems for efficient image storage, transmission, and display. In the acceptance of lossy compression algorithms in the clinical environment, important factors are the assessment of 'visually lossless' compression thresholds, as well as the development of assessment methods requiring fewer data and time than observer performance based studies. In this study a set of quantitative measurements related to medical image quality parameters is proposed for compression assessment. Measurements were carried out using region of interest (ROI) operations on computer-generated test images, with characteristics similar to radiographic images. As a paradigm, the assessment of the lossy Joint Photographic Expert Group (JPEG) algorithm, available in a telematics application for healthcare, is presented. A compression ratio of 15 was found as the visually lossless threshold for the JPEG lossy algorithm, in agreement with previous observer performance studies. Up to this ratio low contrast discrimination is not affected, image noise level is decreased, high contrast line-pair amplitude is decreased by less than 3%, and input/output gray level differences are minor (less than 1%). This type of assessment provides information regarding the type of loss, offering cost and time benefits, in parallel with the advantages of test image adaptation to the requirements of a certain imaging modality and clinical study. PMID- 12758133 TI - A new method to compare planned and achieved position of an orthopaedic implant. AB - The present study describes an automatic method to evaluate the efficacy of a computer aided orthopaedic surgery system by comparing the position of the joint implant, as derived from post-operative computed tomography (CT) scans, to that planned by the surgeon before the operation. The method relies on two spatial registrations, one to align the post-operative femur with the pre-operative femur, the second to compute the planned versus achieved (PVA) accuracy as the roto-translation that registers the pre-operative implant position with the post operative position. Two surface registration algorithms (a generic average distance minimisation and the specialised iterative closest point (ICP) method) were comparatively evaluated first on a set of test cases to measure the absolute accuracy and robustness with respect to peculiar situations such as a distant starting point. The average distance method failed the registration of one test case and showed peak errors of 0.97 degrees on the rotations and 3.09 mm on the translations. The ICP method was found much more efficient and was able to register all test cases. The peak error was 0.44 degrees on the rotations and 0.67 mm on the translations. The ICP method was then used to compute the PVA accuracy on six clinical cases treated with a CT-based planning system in combination with conventional surgical procedures. The method successfully processed all cases demonstrating the efficacy of the proposed procedure in the specific application. PMID- 12758134 TI - An electronic post market surveillance system for medical devices. AB - An important issue in the health care sector is the collection of information about adverse incidents relative to medical devices and the exchange of this information among Health Care Institutions, Manufacturers and Competent Authorities. We present an Electronic Post Market Surveillance System, which solves in an efficient and secure way the above problem and it is compliant to the International Standards. The system consists of the PMS Server Application installed in the manufacturers/suppliers premises and the PMS Client Application installed in the Health Care Institutions. The PMS applications are used for the management of the PMS Reports and Responses with important features such as the user-friendly interfaces, interoperability and different implementations depending on the performance and cost requirements. The messages are in XML format and the security is based on public-private key cryptography. The system was evaluated in a systematic way by a variety of users, who were satisfied by the system functionality and efficiency. PMID- 12758135 TI - Principal component regression analysis with SPSS. AB - The paper introduces all indices of multicollinearity diagnoses, the basic principle of principal component regression and determination of 'best' equation method. The paper uses an example to describe how to do principal component regression analysis with SPSS 10.0: including all calculating processes of the principal component regression and all operations of linear regression, factor analysis, descriptives, compute variable and bivariate correlations procedures in SPSS 10.0. The principal component regression analysis can be used to overcome disturbance of the multicollinearity. The simplified, speeded up and accurate statistical effect is reached through the principal component regression analysis with SPSS. PMID- 12758136 TI - Computing P-values for a class of permutation tests of equal survival functions. AB - Dallas and Rao (Biometrics 56 (2000) 154) proposed a class of permutation tests for testing the equality of two survival distributions based on randomly right censored survival time data consisting of both paired and unpaired observations. Data sets of this type can occur frequently in medical settings. Two members of this class were advocated for use due to their generally high power for detecting scale and location shifts in the exponential and log-logistic distributions for the survival times, and improved power over paired data test procedures that disregard unpaired observations. Because the computations for the tests become quite laborious as the sample sizes increase, computing routines are required for practical implementation of these tests. This paper provides computing routines to execute the tests. PMID- 12758137 TI - Comparing the importance of prognostic factors in Cox and logistic regression using SAS. AB - Two SAS macro programs are presented that evaluate the relative importance of prognostic factors in the proportional hazards regression model and in the logistic regression model. The importance of a prognostic factor is quantified by the proportion of variation in the outcome attributable to this factor. For proportional hazards regression, the program %RELIMPCR uses the recently proposed measure V to calculate the proportion of explained variation (PEV). For the logistic model, the R(2) measure based on squared raw residuals is used by the program %RELIMPLR. Both programs are able to compute marginal and partial PEV, to compare PEVs of factors, of groups of factors, and even to compare PEVs of different models. The programs use a bootstrap resampling scheme to test differences of the PEVs of different factors. Confidence limits for P-values are provided. The programs further allow to base the computation of PEV on models with shrinked or bias-corrected parameter estimates. The SAS macros are freely available at www.akh-wien.ac.at/imc/biometrie/relimp PMID- 12758138 TI - Distortion correction for digital subtraction angiography imaging: PC based system for radiosurgery planning. AB - We report, the development of a personal computer (PC) based system to correct distortion of digital subtraction angiography (DSA) images. The program was written in INTERACTIVE DATA LANGUAGE (IDL) and implemented on a PC equipped with an Intel Pentium III 450 MHz CPU in the MICROSOFT WINDOWS 98 environment. The system consists of two modules. The coefficient calculation module detects distortions of grid phantom images automatically and determines the distortion correction function. An additional distortion correction module corrects detected distortion using the correction function determined by the coefficient calculation module. The correction program can be used for images taken at arbitrary lateral oblique angle, and about 4 min are required to correct an image. The correction program was verified using phantom and clinical images. After image correction, the root mean square (rms) deviations of the reference points of each image were calculated. The average value of the rms deviation of all phantom images was about 0.1 mm. The residual mean rms deviation of the corrected clinical images (0.34+/-0.19 mm) and maximum error (0.59+/-0.26 mm) were within the acceptable limits of stereotactic radiosurgery. The accuracy, the ability to process lateral oblique angles, and reasonable program running time makes the developed system a valuable tool in clinical practice, for example for the planning of gamma knife radiosurgery. PMID- 12758139 TI - Np.OR: an S-Plus function for pointwise nonparametric estimation of odds-ratios of continuous predictors. AB - Calculating odds ratios (OR) and corresponding confidence intervals (CI) for exposures that have been measured using a continuous scale, presents important limitations in the traditional practice of epidemiology. Our objective is to describe an S-Plus function, that we called np.OR, that allows the computation of the pointwise estimates of the ORs as well as their corresponding CIs of continuous predictors introduced nonlinearly in a generalised model. The function can also be generalised to compute the pointwise estimates of categorical predictors that were introduced nonlinearly. To illustrate usage of the program we analyse the relationship between ambient temperature and total mortality in Barcelona for the period 1991-1995, while controlling for observed and unobserved confounders. PMID- 12758140 TI - Fixing the nonconvergence bug in logistic regression with SPLUS and SAS. AB - When analyzing clinical data with binary outcomes, the parameter estimates and consequently the odds ratio estimates of a logistic model sometimes do not converge to finite values. This phenomenon is due to special conditions in a data set and known as 'separation'. Statistical software packages for logistic regression using the maximum likelihood method cannot appropriately deal with this problem. A new procedure to solve the problem has been proposed by Heinze and Schemper (Stat. Med. 21 (2002) pp. 2409-3419). It has been shown that unlike the standard maximum likelihood method, this method always leads to finite parameter estimates. We developed a SAS macro and an SPLUS library to make this method available from within one of these widely used statistical software packages. Our programs are also capable of performing interval estimation based on profile penalized log likelihood (PPL) and of plotting the PPL function as was suggested by Heinze and Schemper (Stat. Med. 21 (2002) pp. 2409-3419). PMID- 12758141 TI - Crystallographic structure at 1.6-A resolution of the human adenovirus proteinase in a covalent complex with its 11-amino-acid peptide cofactor: insights on a new fold. AB - The crystal structure of the human adenovirus proteinase (AVP), a cysteine proteinase covalently bound to its 11-amino-acid peptide cofactor pVIc, has been solved to 1.6-A resolution with a crystallographic R-factor of 0.136, R(free)=0.179. The fold of AVP-pVIc is new and the structural basis for it is described in detail. The polypeptide chain of AVP folds into two domains. One domain contains a five-strand beta-sheet with two peripheral alpha-helices; this region represents the hydrophobic core of the protein. A second domain contains the N terminus, several C-terminal alpha-helices, and a small peripheral anti parallel beta-sheet. The domains interact through an extended polar interface. pVIc spans the two domains like a strap, its C-terminal portion forming a sixth strand on the beta-sheet. The active site is in a long, deep groove located between the two domains. Portions are structurally similar to the active site of the prototypical cysteine proteinase papain, especially some of the Calpha backbone atoms (r.m.s. deviation of 0.354 A for 12 Calpha atoms). The active-site nucleophile of AVP, the conserved Cys(122), was shown to have a pK(a) of 4.5, close to the pK(a) of 3.0 for the nucleophile of papain, suggesting that a similar ion pair arrangement with His(54) may be present in AVP-pVIc. The interactions between AVP and pVIc include 24 non-beta-strand hydrogen bonds, six beta-strand hydrogen bonds and one covalent bond. Of the 204 amino acid residues in AVP, 33 are conserved among the many serotypes of adenovirus, and these aid in forming the active site groove, are involved in substrate specificity or interact between secondary structure elements. PMID- 12758142 TI - Site-directed fluorescence probing to dissect the calcium-dependent association between soluble tissue factor and factor VIIa domains. AB - We have used the site-directed labeling approach to study the Ca(2+)-dependent docking of factor VIIa (FVIIa) to soluble tissue factor (sTF). Nine Ca(2+) binding sites are located in FVIIa and even though their contribution to the overall binding between TF and FVIIa has been thoroughly studied, their importance for local protein-protein interactions within the complex has not been determined. Specifically we have monitored the association of the gamma carboxyglutamic acid (Gla), the first EGF-like (EGF1), and the protease domains (PD) of FVIIa to sTF. Our results revealed that complex formation between sTF and FVIIa during Ca(2+) titration is initiated upon Ca(2+) binding to EGF1, the domain containing the site of highest Ca(2+) affinity. Besides we showed that a Ca(2+)-loaded Gla domain is required for an optimal association of all domains of FVIIa to sTF. Ca(2+) binding to the PD seems to be of some importance for the docking of this domain to sTF. PMID- 12758143 TI - Expression of recombinant human L-glutaminase in Escherichia coli: polyclonal antibodies production and immunological analysis of mouse tissues. AB - The first complete sequence of human L-glutaminase was deduced from breast cancer glutaminase cDNA cloned in our laboratory. This cDNA clone has now been engineered to synthesize both precursor and mature forms of the protein in Escherichia coli. Among several different plasmid constructions, the expression system based on phage T7 promoter (vector pET-3c) was found to be the most efficient for glutaminase overproduction. Upon induction, precursor glutaminase accounts for about 25% of total E. coli protein, whereas a lower amount (12%) was achieved for the putative mature protein. The optimal length of the translational spacer on the ribosome binding site was shown to be eight nucleotides. However, using this length of spacer, we were unable to obtain expression in the pQE vector, tagged with a 6x His sequence at the NH(2)-terminus, stressing the importance of the 5'-coding sequence in the expression efficiency. Although the precursor and mature recombinant forms of glutaminase were devoid of catalytic activity, the purified protein allowed us to obtain highly specific polyclonal antibodies, as shown by immunoblot analysis of mouse tissues. Furthermore, the antibodies were able to immunoprecipitate the in vitro translated enzyme using a reticulocyte lysate system; these antibodies might be a valuable tool for studies on L-glutaminase expression in mammalian tissues. PMID- 12758144 TI - Alcoholysis catalyzed by Candida antarctica lipase B in a gas/solid system: effects of water on kinetic parameters. AB - The influence of water on the kinetics of alcoholysis of methyl propionate and n propanol catalyzed by immobilized lipase B from Candida antarctica was studied in a continuous solid/gas reactor. In this reactor, the solid phase is composed of a packed enzymatic sample which is percolated by gaseous nitrogen, simultaneously carrying gaseous substrates to the enzyme while removing reaction products. In this system, interactions between the enzyme and nonreacting molecules are avoided, since no solvent is present, and it is thus more easy to assess the role of water. To this end, alcohol inhibition constant, substrates dissociation constants as well as acylation rate constant and ratio of acylation to deacylation rate constants have been determined as a function of water activity (a(w)). Data obtained highlight that n-propanol inhibition constant and dissociation constant of methyl propionate are a lot affected by a(w) variations whereas water has no significant effect on the catalytic acylation step nor on the ratio of acylation to deacylation rate constants. These results suggest the water-independent character of the transition step. PMID- 12758145 TI - Multiple isoforms of choline kinase from Caenorhabditis elegans: cloning, expression, purification, and characterization. AB - Choline kinase is the first enzymatic step in the CDP-choline pathway for phosphatidylcholine biosynthesis. The genome of the nematode, Caenorhabditis elegans, contains seven genes that appear likely to encode choline and/or ethanolamine kinases. We cloned five and expressed four of these genes, and purified or partially purified three of the encoded enzymes. All expressed proteins had choline kinase activity; those that most closely resemble the mammalian choline kinases were the most active. CKA-2, a very active form, was purified to near homogeneity. The K(m) values for CKA-2 were 1.6 and 2.4 mM for choline and ATP, respectively, and k(cat) was 74 s(-1). CKA-2 was predominantly a homodimer as assessed by glycerol gradient sedimentation and dynamic light scattering. CKB-2, which was less similar to mammalian choline kinases, had K(m) values for choline and ATP of 13 and 0.7 mM, and k(cat) was 3.8 s(-1). Both of these highly purified enzymes required magnesium, had very alkaline pH optima, and were much more active with choline as substrate than with ethanolamine. These results provide a foundation for future studies on the structure and function of choline kinases, as well as studies on the genetic analysis of the function of the multiple isoforms in this organism. PMID- 12758146 TI - Effects of salts on the function and conformational stability of shikimate kinase. AB - The unfolding of shikimate kinase (SK) from Erwinia chrysanthemi by urea and its subsequent refolding on dilution of the denaturing agent has been studied in detail [Eur. J. Biochem. 269 (2002) 2124]. Comparison of the effects of urea on the enzyme with those of guanidinium chloride (GdmCl) and NaCl indicated that chloride ions significantly weakened the binding of shikimate. This finding prompted a more detailed examination of the effects of salts on the structure, function and stability of the enzyme; the effects of NaCl and Na(2)SO(4) were investigated in detail. These salts have very small effects on the secondary structure as judged by far UV CD circular dichroism (CD), although the near UV CD spectra suggest that some limited changes in the environment of aromatic amino acids may occur. Both salts inhibit SK activity and analysis of the kinetic and substrate binding parameters point to a complex mechanism for the inhibition. Inclusion of salts leads to a marked stabilisation against unfolding of the enzyme by urea. When the enzyme is unfolded by incubation in 4 M urea, addition of NaCl or Na(2)SO(4) leads to a relatively slow refolding of the enzyme as judged by the regain of native-like CD and fluorescence. In addition, the refolded enzyme can bind shikimate, though more weakly than the native enzyme. However, the refolded enzyme does not appear to be capable of binding nucleotides, nor does it possess detectable catalytic activity. The refolding process brought about by addition of salt in the presence of 4 M urea is not associated with any change in the fluorescence of the probe 8-anilino-1 naphthalenesulfonic acid (ANS), indicating that an intermediate formed by hydrophobic collapse is unlikely to be significantly populated. The results point to both specific and general effects of salts on SK. These are discussed in the light of the structural information available on the enzyme. PMID- 12758147 TI - Kohamaic acid A, a novel sesterterpenic acid, inhibits activities of DNA polymerases from deuterostomes. AB - We previously found and isolated a novel natural product, designated kohamaic acid A (KA-A), which inhibited the first cleavage of fertilized sea urchin eggs. In this paper, we report that this compound could selectively inhibit the activities of DNA polymerases (pol. alpha, beta, gamma, delta and epsilon ) only from species in the deuterostome branch in the animal kingdom, like sea urchin, fish and mammals, but not from protostomes including insects (fruit fly, Drosophila melanogaster) and mollusks (octopus and oyster). Inhibition of deuterostome DNA polymerases was dose dependent. IC(50) values for DNA polymerases of mammals and fish occurred at approximately 5.8-14.9 microM and those of sea urchin at 6.1-30.3 microM. In the sea urchin DNA polymerases, the activities of the replicative DNA polymerases such as alpha, delta and epsilon were more strongly inhibited than that of the repair-related pol. beta. KA-A is an inhibitor of replicative DNA polymerases from the deuterostome species, and subsequently, the inhibition of the first cleavage of fertilized sea urchin eggs might occur as a result of the suppression of DNA replication. PMID- 12758148 TI - Soluble human core 2 beta6-N-acetylglucosaminyltransferase C2GnT1 requires its conserved cysteine residues for full activity. AB - Human UDP-GlcNAc: Galbeta1-3GalNAc- (GlcNAc to GalNAc) beta1,6-GlcNAc-transferase (C2GnT1) is a member of a group of beta6-GlcNAc-transferases that belongs to CAZy family 14. One of the striking features of these beta6-GlcNAc-transferases is the occurrence of nine completely conserved cysteine residues that are located throughout the catalytic domain. We have expressed the soluble catalytic domain of human C2GnT1 in insect cells, and isolated active enzyme as a secreted protein. beta-Mercaptoethanol (beta-ME) and dithiothreitol (DTT) were found to stimulate the enzyme activity up to 20-fold, indicating a requirement for a reduced sulfhydryl for activity. When the enzyme was subjected to nonreducing PAGE, the migration of the protein was identical to the migration in reducing gels, demonstrating the absence of intermolecular disulfide bonds. This suggested that the monomer is the active form of the enzyme. Sulfhydryl reagents such as 5,5'-dithiobis-2-nitrobenzoic acid (DTNB) and N-ethylmaleimide (NEM) inactivated the enzyme, and the inactivation was partially prevented by prior addition of donor or acceptor substrate and by sulfhydryl reducing agents. We therefore investigated the role of all nine conserved cysteine residues in enzyme stability and activity by site-directed mutagenesis where individual cysteine residues were changed to serine. All of the mutants were expressed as soluble proteins. Seven of the Cys mutants were found to be inactive, while C100S and C217S mutants had 10% and 41% activity, respectively, when compared to the wild-type enzyme. Wild type and C217S enzymes had similar K(M) and V(max) values for acceptor substrate Galbeta1-3GalNAcalpha-p-nitrophenyl (GGApnp), but the K(M) value for UDP-GlcNAc was higher for C217S than for the wild-type enzyme. In contrast to wild-type enzyme, C217S was not stimulated by reducing agents and was not inhibited by sulfhydryl specific reagents. These results suggest that Cys-217 is a free sulfhydryl in active wild-type enzyme and that Cys-217, although not required for activity, is in or near the active site of the protein. Since seven of the mutations were totally inactive, it is likely that these seven Cys residues play a role in maintaining an active conformation of soluble C2GnT1 by forming disulfide bonds. These bonds are only broken at high concentrations of disulfide reducing agents. PMID- 12758149 TI - Recombinant human C1-inhibitor produced in Pichia pastoris has the same inhibitory capacity as plasma C1-inhibitor. AB - Therapeutic application of the serpin C1-inhibitor (C1-Inh) in inflammatory diseases like sepsis, acute myocardial infarction and vascular leakage syndrome seems promising, but large doses may be required. Therefore, a high-yield recombinant expression system for C1-Inh is very interesting. Earlier attempts to produce high levels of C1-Inh resulted in predominantly inactive C1-Inh. We describe the high yield expression of rhC1-Inh in Pichia pastoris, with 180 mg/l active C1-Inh at maximum. On average, 30 mg/l of 80-100% active C1-Inh was obtained. Progress curves were used to study the interaction with C1s, kallikrein, coagulation factor XIIa and XIa, and demonstrated that rhC1-Inh had the same inhibitory capacity as plasma C1-Inh. Structural integrity, as monitored via heat stability, was comparable despite differences in extent and nature of glycosylation. We conclude that the P. pastoris system is capable of high-level production of functionally and structurally intact human C1 inhibitor. PMID- 12758150 TI - Structural and functional characterisation of the DNA binding domain of the Aspergillus nidulans gene regulatory protein AreA. AB - The 876-aa protein AreA regulates the expression of numerous genes involved in nitrogen metabolism in Aspergillus nidulans, and interacts with GATA sequences upstream of the relevant genes. We have carried out limited proteolysis of the C terminal domain of the AreA protein in order to identify possible structural domains within the protein. A stable 156-amino-acid fragment was identified that contained the zinc finger region, and this sequence was cloned and expressed in E. coli. Fluorescence spectroscopy of the purified protein showed that the proteolytic domain was folded and could be denatured by high concentrations of urea (approximately 4 M), exhibiting a sharp transition. Fluorescence spectroscopy was also used to monitor binding to a DNA duplex containing the AreA recognition site, demonstrating tight binding of the domain to its DNA recognition sequence. The DNA binding affinity of the domain is comparable with that of the native AreA protein and much higher than that of the minimal zinc finger region of AreA. PMID- 12758151 TI - Cloning and expression of calglandulin, a new EF-hand protein from the venom glands of Bothrops insularis snake in E. coli. AB - The EF-hand protein family is comprised of many proteins with conserved Ca(2+) binding motifs with important biological roles in intracellular communication. During the generation of Expressed Sequence Tags (ESTs) from the venom glands of the Viperidae snake Bothrops insularis, we identified a cDNA coding for a putative Ca(2+) binding protein with four EF-hand motifs, named here calglandulin. The deduced amino acid sequence displayed the greatest sequence similarity with calmodulin (59%), followed by troponin-C (52%). The encoded polypeptide was first expressed in Escherichia coli as a 6XHis-tagged fusion protein. The expressed protein was purified by Ni(2+)-charged affinity chromatography and circular dichroism (CD) spectroscopy confirmed the prevalence of alpha-helix as observed in calmodulin/calmodulin-like proteins. A polyclonal antiserum was generated in mice using this recombinant calglandulin. To investigate the tissue-specific biological occurrence of this protein, this antiserum was used in Western blot experiments, which revealed an immunoreactive band in samples of venom gland extracts from different snakes, but not in the crude venom or in brain, heart and other tissues. This exclusive occurrence suggests a specialized function of calglandulin in snake venom glands. PMID- 12758152 TI - Creation of soybean beta-conglycinin beta with strong phagocytosis-stimulating activity. AB - beta-Conglycinin is composed of three kinds of subunit: alpha, alpha' and beta. A phagocytosis-stimulating peptide sequence (MITLAIPVNKPGR), soymetide, exists in the alpha' subunit of beta-conglycinin. Met at N terminus of the soymetide is essential for the activity. When Thr at the third residue from N terminus of the soymetide is replaced by Phe or Trp, the phagocytosis-stimulating activity greatly increases (ThrMet, Lys- >Thr, Phe, or Trp) into the beta subunit after confirmation of the effects of residue replacements by molecular modeling, suggesting that the introduced mutations might not prevent the correct folding. The studies of circular dichroism (CD), gel filtration and differential scanning calorimetry (DSC) of the mutants (I122M/K124T, I122M/K124F, I122M/K124W) expressed in E. coli demonstrated that they folded and self-assembled similarly to the wild type. This was confirmed by X-ray analysis of I122M/K124W crystal where the biggest residue tryptophane was introduced. The three mutants exhibited phagocytosis activities after digestion by trypsin, and the order was the wild type95% accuracy. The RNA- and DNA binding SVMs demonstrate more diverse accuracy, ranging from approximately 76% to approximately 97%. Analysis of the test results suggests the directions of improving the SVMs. PMID- 12758156 TI - Spectrofluorimetric analysis of 7-hydroxycoumarin binding to bovine phenol sulfotransferase. AB - Phenol sulfotransferases (SULT1s, EC 2.8.2.1) catalyze sulfuryl group transfer from 3'-phosphoadenosine-5'-phosphosulfate (PAPS) to the hydroxyl oxygen of aromatic acceptor substrates. Previous work with the bovine SULT1A1 has utilized the highly fluorescent substrate 7-hydroxycoumarin (7-HC, umbelliferone) as an acceptor substrate [Biochem. Biophys. Res. Commun. 261 (1999) 815]. Here we report that adenosine-3',5'-bisphosphate (PAP)-dependent binding of 7-HC to bSULT1A1 can be observed due to the appearance of a 400-420-nm shoulder in the emission spectrum, using an excitation wavelength of 280 nm. This emission was observed by placing 7-HC in ethanol, which is consistent with bSULT1A1 phenol binding site hydrophobicity. Titrations with 7-HC indicate a K(d) for 7-HC of 0.58 microM and substoichiometric binding to the homodimeric enzyme. The bSULT1A1:PAP:7-HC complex could be disrupted with pentachlorophenol (PCP), titrations with which indicated 0.5 equivalents per enzyme subunit. Titrations of enzyme plus 7-HC with PAP also indicated 0.5 equivalents per enzyme subunit. These results suggest a model of homodimeric bSULT1A1 in which subunit interactions favor half-site reactivity in the formation of a dead end complex. PMID- 12758157 TI - Digestion of mu- and m-calpain by trypsin and chymotrypsin. AB - Proteolytic digestion by trypsin and chymotrypsin was used to probe conformation and domain structure of the mu- and m-calpain molecules in the presence and the absence of Ca(2+). Both calpains have a compact structure in the absence of Ca(2+); incubation with either protease for 120 min results in only three or four major fragments. A 24-kDa fragment was produced by removal of the Gly-rich area in domain V of the 28-kDa subunit. The other fragments were from the 80-kDa subunit. Except for trypsin digestion of m-calpain, the region between amino acids 245 and 265 (human sequence) was very susceptible to cleavage by both proteases in the absence of Ca(2+); this region is in domain II (IIb of the crystallographic structure). Although no proteolytically active fragments could be isolated from either tryptic or chymotryptic digests, the calpain molecule can remain assembled in a proteolytically active complex even after the 80-kDa subunit has been completely degraded. The results suggest that interaction among different regions of the entire calpain molecule is required for its full proteolytic activity. In the presence of 1 mM Ca(2+), both calpains are degraded to fragments less than 40-kDa in less than 5 min. The C-terminal ends of both subunits, from amino acids 503 to 506 to the end of the 80-kDa subunit and from amino acids 85 to 88 to the end of the 28-kDa subunit, were resistant to degradation by either protease in the presence or in the absence of Ca(2+). Hence, this part of the calpain molecule is in a compact structure that does not change significantly in the presence of Ca(2+). PMID- 12758158 TI - Recombinant human antibodies against the reverse transcriptase of human immunodeficiency virus type-1. AB - Inhibitory antibodies to the reverse transcriptase (RT) of human immunodeficiency virus type-1 (HIV-1) can be used to block the life cycle of the virus. We have isolated five different human single chain Fv (ScFv) antibodies specific for HIV 1 RT from an antibody phage display library. Three of these antibodies inhibited the RNA-dependent DNA polymerase (RDDP) activity of RT and one of the three (F-6) inhibited also its DNA-dependent DNA polymerase (DDDP) activity. Unexpectedly, F 6 binds to the carboxyl terminus of the large subunit of RT, which contains the ribonuclease H (RNase H) domain, and not the polymerase domain of the protein. Moreover, this binding did not inhibit the RNase H enzymatic activity. To further characterize F-6 antibody, two cyclic synthetic peptides based on the amino acids sequences of the CDR3 of F-6 were synthesized. Peptide F-6CDRH3, with the sequence of CDR3 of the heavy chain, inhibited the RDDP activity of RT while peptide F-6CDRL3, with the sequence of CDR3 of the light chain, had no effect on this activity of RT. These results indicate that some of the effects of F-6 are mediated by the CDR3 of the heavy chain. The antibodies identified here will be further tested as intrabodies for their capacity to protect human cells from HIV 1 infection. PMID- 12758159 TI - Identification of an equilibrium intermediate in the unfolding process of galectin-1, which retains its carbohydrate-binding specificity. AB - The unfolding process of galectin-1 (Gal-1) in the presence of a denaturing agent was examined using fluorescence and far-UV circular dichroism (CD) spectroscopy determinations, and was found to be completely reversible. The data showed that the transitions of guanidine hydrochloride (GdnHCl)-induced lectin unfolding, in the absence of ligand, were biphasic in nature, clearly showing the existence of at least one stable intermediate. On the other hand, the unfolding in the presence of disaccharide yielded data that could fit very well to a two-state model, indicating a stabilizing effect of the ligand. The folding intermediate was further characterized by size exclusion chromatography, near-UV CD and anilinonaphtalene sulfonate binding, and shown to belong to the molten globule type. Strikingly, this intermediate retained its carbohydrate-binding specificity, as evidenced by the tryptophan fluorescence changes detected upon its interaction with lactose. PMID- 12758160 TI - Denaturant-induced equilibrium unfolding of concanavalin A is expressed by a three-state mechanism and provides an estimate of its protein stability. AB - The urea and guanidine hydrochloride (GdnHCl)-induced denaturation of tetrameric concanavalin A (ConA) at pH 7.2 has been studied by using intrinsic fluorescence, 8-anilino-1-naphthalenesulfonate (ANS) binding, far-UV circular dichroism (CD), and size-exclusion chromatography. The equilibrium denaturation pathway of ConA, as monitored by steady state fluorescence, exhibits a three-state mechanism involving an intermediate state, which has been characterized as a structured monomer of the protein by ANS binding, far-UV CD and gel filtration size analysis. The three-state equilibrium is analyzed in terms of two distinct and separate dissociation (native tetramer<-->structured monomer) and unfolding (structured monomer<-->unfolded monomer) reaction steps, with the apparent transition midpoints (C(m)), respectively, at 1.4 and 4.5 M in urea, and at 0.8 and 2.4 M in GdnHCl. The results show that the free energy of stabilization of structured monomer relative to the unfolded state (-DeltaG(unf, aq)), is 4.4-5.5 kcal mol(-1), and that of native tetramer relative to structured monomer ( DeltaG(dis, aq)) is 7.2-7.4 kcal mol(-1), giving an overall free energy of stabilization (-DeltaG(dis&unf, aq)) of 11.6-12.9 kcal mol(-1) (monomer mass) for the native protein. However, the free energy preference at the level of quaternary tetrameric structure is found to be far greater than that at the tertiary monomeric level, which reveals that the structural stability of ConA is maintained mostly by subunit association. PMID- 12758161 TI - Equilibrium unfolding of RNase Rs from Rhizopus stolonifer: pH dependence of chemical and thermal denaturation. AB - The conformational stability of RNase Rs was determined with chemical and thermal denaturants over the pH range of 1-10. Equilibrium unfolding with urea showed that values of D(1/2) (5.7 M) and DeltaG(H(2)O) (12.8 kcal/mol) were highest at pH 5.0, its pI and the maximum conformational stability of RNase Rs was observed near pH 5.0. Denaturation with guanidine hydrochloride (GdnHCl), at pH 5.0, gave similar values of DeltaG(H(2)O) although GdnHCl was 2-fold more potent denaturant with D(1/2) value of 3.1 M. The curves of fraction unfolded (f(U)) obtained with fluorescence and CD measurements overlapped at pH 5.0. Denaturation of RNase Rs with urea in the pH range studied was reversible but the enzyme denatured irreversibly >pH 11.0. Thermal denaturation of RNase Rs was reversible in the pH range of 2.0-3.0 and 6.0-9.0. Thermal denaturation in the pH range 4.0-5.5 resulted in aggregation and precipitation of the protein above 55 degrees C. The aggregate was amorphous or disordered precipitate as observed in TE micrographs. Blue shift in emission lambda(max) and enhancement of fluorescence intensity of ANS at 70 degrees C indicated the presence of solvent exposed hydrophobic surfaces as a result of heat treatment. Aggregation could be prevented partially with alpha-cyclodextrin (0.15 M) and completely with urea at concentrations >3 M. Aggregation was probably due to intermolecular hydrophobic interaction favored by minimum charge-charge repulsion at the pI of the enzyme. Both urea and temperature-induced denaturation studies showed that RNase Rs unfolds through a two-state F right arrow over left arrow U mechanism. The pH dependence of stability described by DeltaG(H(2)O) (urea) and DeltaG (25 degrees C) suggested that electrostatic interactions among the charged groups make a significant contribution to the conformational stability of RNase Rs. Since RNase Rs is a disulfide-containing protein, the major element for structural stability are the covalent disulfide bonds. PMID- 12758162 TI - Tight-binding inhibition by alpha-naphthoflavone of human cytochrome P450 1A2. AB - Human cytochrome P450 (P450) enzymes exhibit remarkable diversity in their substrate specificities, participating in oxidation reactions of a wide range of xenobiotic drugs. Previously, we reported that alpha-naphthoflavone (ANF) is bound to the recombinant P450 1A2 tightly and stabilizes an overall enzyme conformation. The present study is designed to determine the type of P450 1A2 inhibition exerted by ANF, using two different substrates of P450 1A2, 7 ethoxycoumarin (EOC) and 7-ethoxyresorufin (EOR). ANF is generally known as a competitive inhibitor of the enzyme. However, in our tight-binding enzyme kinetics study, ANF acts as noncompetitive inhibitor in 7-ethoxycoumarin O deethylation (ECOD) (K(i)=55.0 nM), but as competitive inhibitor in 7 ethoxyresorufin O-deethylation (EROD) (K(i)=1.4 nM). Based on homology modeling studies, ANF is positioned to bind to a hydrophobic cavity next to the active site where it may cause a direct effect on substrate binding. It is agreed with the predicted binding site of ANF in P450 3A4, in which ANF is rather known as a stimulating modulator. Our results suggest that ANF binds near the active site of P450 1A2 and exhibits differential inhibition mechanisms, possibly depending on the molecular structure of the substrate. PMID- 12758163 TI - The molecular class C acid phosphatase of Chryseobacterium meningosepticum (OlpA) is a broad-spectrum nucleotidase with preferential activity on 5'-nucleotides. AB - The olpA gene of Chryseobacterium meningosepticum, encoding a molecular class C phosphatase, was cloned and expressed in Escherichia coli. The gene encodes a 29 kDa polypeptide containing an amino-terminal signal peptide typical of bacterial membrane lipoproteins. Expression in E. coli results in a functional product that mostly partitions in the outer membrane. A secreted soluble OlpA derivative (sOlpA) lacking the N-terminal cysteine residue for lipid anchoring was produced in E. coli and purified by means of two steps of ion exchange chromatography. Analysis of the kinetic parameters of sOlpA with several organic phosphoesters revealed that the enzyme was able to efficiently hydrolyze nucleotide monophosphates, with a strong preference for 5'-nucleotides and for 3'-AMP. The enzyme was also able to hydrolyze sugar phosphates and beta-glycerol phosphate, although with a lower efficiency, whereas it was apparently inactive against nucleotide di- and triphosphates, diesters, and phytate. OlpA, therefore, can be considered a broad-spectrum nucleotidase with preference for 5'-nucleotides. Its functional behaviour exhibits differences from that of the Haemophilus influenzae OMP P4 lipoprotein, revealing functional heterogeneity among phosphatases of molecular class C. PMID- 12758165 TI - Intracellular warfare between human influenza viruses and human cells: the roles of the viral NS1 protein. PMID- 12758164 TI - Heterogeneity in gamma-glutamyltransferase mRNA expression and glycan structures. Search for tumor-specific variants in human liver metastases and colon carcinoma cells. AB - The enzyme gamma-glutamyltransferase (GGT) is frequently overexpressed in cancer cells and tissues and has significant utility as a cancer marker. Significant heterogeneity of the enzyme has been described due to both transcriptional and post-translational variations. For possible use in diagnosis and follow-up of patients with colorectal cancer, a search was performed for specific mRNA subtypes and glycan structures of the enzyme in liver metastases. We found no differences in the distribution of three GGT mRNA subtypes (fetal liver, HepG2, placenta) in metastatic tissue and normal liver tissue. Furthermore, the three subtypes were present in leukocytes isolated from both normal individuals and cancer patients. Two colon carcinoma cell lines (Colo 205 and HCC 2998) also displayed the three forms and no consistent changes in mRNA composition were noted after butyrate-induced differentiation of the cells. Thus, neither of the GGT mRNA subforms appear to be tumor-specific, although some qualitative and quantitative variations were noted. Two distinct glycosylation features were detected for GGT in metastatic tissue in contrast to normal liver GGT; an extreme sialic acid heterogeneity and a significant increase in beta1,6GlcNAc branching. The GGT glycans from the two colon carcinoma cell lines also possessed these features. As butyrate treatment of the cells resulted in an increased sialic acid content and a reduced beta1,6GlcNAc branching, the described carbohydrate structures appear to be part of a tumor-related pattern. We were, however, unable to identify such GGT isoforms in serum from patients with advanced colorectal cancer. This indicates that their usefulness in diagnostic use is doubtful. PMID- 12758166 TI - Limited evolution of West Nile virus has occurred during its southwesterly spread in the United States. AB - Analysis of partial nucleotide sequences of nine West Nile virus strains isolated in southeast Texas during June-August 2002 revealed a maximum of 0.35% nucleotide variation from a New York 1999 strain. Two sequence subtypes were identified that differed from each other by approximately 0.5%, suggesting multiple introductions of virus to this area. Analysis of sequences from cloned PCR products for one strain revealed up to 0.6% divergence from the consensus sequence at the subpopulation level. The presence of unique patterns of small numbers of mutations in North American West Nile strains studied to date may suggest the absence of a strong selective pressure to drive the emergence of dominant variants. PMID- 12758167 TI - Regulation of MCP-1 gene transcription by Smads and HIV-1 Tat in human glial cells. AB - Expression of several cytokines involved in signal transduction such as TGFbeta-1 and the inflammatory chemokines including MCP-1 is elevated during the course of AIDS progression. The enhancement of these cellular proteins in astrocytic cells is mediated, at least in part, by HIV-1 Tat protein. Here, we investigate the possible regulation of MCP-1 transcription by Tat and the Smad family of transcription factors whose activities are induced by the TGFbeta-1 pathway. Results from transfection studies revealed that Smad-3 stimulates basal and Tat mediated transcription of MCP-1 in human astrocytic cells. Smad-4, on the other hand, had no effect on the basal activity of the MCP-1 promoter, but showed the ability to decrease both Smad-3 and Tat-induced transcription of the MCP promoter. Results from protein-binding studies revealed the ability of both Smad 3 and Smad-4 to associate with the region of Tat spanning residues 1-40. Examination of the transcriptional activity of the various domains of Smad including MH1, at the N-terminus, and MH2, at the C-terminus of the protein indicated that neither MH1 or MH2 alone positively cooperate with Tat in modulating MCP-1 transcription. However, ectopic expression of MH1 and, more notably, MH2 severely suppressed transcriptional activation of MCP-1 by Tat in astrocytic cells. Binding studies revealed that similar to the full-length Smad protein, both MH1 and MH2 associate with Tat protein and that the residues between 1 and 40 of Tat are important for their interaction. These observations reveal a novel mechanism for Tat-mediated transcriptional activation via TGFbeta signaling pathway and provide evidence for regulation of MCP-1 gene transcription by this signaling pathway in human astrocytic cells. PMID- 12758168 TI - Deleterious effect of peptide insertions in a permissive site of the AAV2 capsid. AB - A permissive site for insertion of heterologous peptide sequences has been identified in the capsid proteins of AAV2. While attempting to use this site for insertion of a nuclear localization sequence, we have observed a drastic reduction in the yield of DNA-containing particles. ELISA analysis showed that capsid assembly was modestly affected, whereas genome encapsidation was more profoundly altered, a phenomenon we did not observed when a RGD peptide was inserted at the same location. Furthermore, the NLS viruses displayed poor transduction efficiency on HeLa and 293 cells. Altogether, these results indicate that the nature of the peptide inserted at position 587 in the capsid may have important consequences on both particle formation and infectivity. PMID- 12758169 TI - X-ray structure of the hemagglutinin of a potential H3 avian progenitor of the 1968 Hong Kong pandemic influenza virus. AB - We have determined the structure of the HA of an avian influenza virus, A/duck/Ukraine/63, a member of the same antigenic subtype, H3, as the virus that caused the 1968 Hong Kong influenza pandemic, and a possible progenitor of the pandemic virus. We find that structurally significant differences between the avian and the human HAs are restricted to the receptor-binding site particularly the substitutions Q226L and G228S that cause the site to open and residues within it to rearrange, including the conserved residues Y98, W153, and H183. We have also analyzed complexes formed by the HA with sialopentasaccharides in which the terminal sialic acid is in either alpha2,3- or alpha2,6-linkage to galactose. Comparing the structures of complexes in which an alpha2,3-linked receptor analog is bound to the H3 avian HA or to an H5 avian HA leads to the suggestion that all avian influenza HAs bind to their preferred alpha2,3-linked receptors similarly, with the analog in a trans conformation about the glycosidic linkage. We find that alpha2,6-linked analogs are bound by both human and avian HAs in a cis conformation, and that the incompatibility of an alpha2,6-linked receptor with the alpha2,3-linkage-specific H3 avian HA-binding site is partially resolved by a small change in the position and orientation of the sialic acid. We discuss our results in relation to the mechanism of transfer of influenza viruses between species. PMID- 12758170 TI - C-terminal region of herpes simplex virus ICP8 protein needed for intranuclear localization. AB - The herpes simplex virus single-stranded DNA-binding protein, ICP8, localizes initially to structures in the nucleus called prereplicative sites. As replication proceeds, these sites mature into large globular structures called replication compartments. The details of what signals or proteins are involved in the redistribution of viral and cellular proteins within the nucleus between prereplicative sites and replication compartments are poorly understood; however, we showed previously that the dominant-negative d105 ICP8 does not localize to prereplicative sites and prevents the localization of other viral proteins to prereplicative sites (J. Virol. 74 (2000) 10122). Within the residues deleted in d105 (1083 to 1168), we identified a region between amino acid residues 1080 and 1135 that was predicted by computer models to contain two alpha-helices, one with considerable amphipathic nature. We used site-specific and random mutagenesis techniques to identify residues or structures within this region that are required for proper ICP8 localization within the nucleus. Proline substitutions in the predicted helix generated ICP8 molecules that did not localize to prereplicative sites and acted as dominant-negative inhibitors. Other substitutions that altered the charged residues in the predicted alpha-helix to alanine or leucine residues had little or no effect on ICP8 intranuclear localization. The predicted alpha-helix was dispensable for the interaction of ICP8 with the U(L)9 origin-binding protein. We propose that this C-terminal alpha helix is required for localization of ICP8 to prereplicative sites by binding viral or cellular factors that target or retain ICP8 at specific intranuclear sites. PMID- 12758171 TI - Herpes simplex virus replication compartments can form by coalescence of smaller compartments. AB - Herpes simplex virus (HSV) uses intranuclear compartmentalization to concentrate the viral and cellular factors required for the progression of the viral life cycle. Processes as varied as viral DNA replication, late gene expression, and capsid assembly take place within discrete structures within the nucleus called replication compartments. Replication compartments are hypothesized to mature from a few distinct structures, called prereplicative sites, that form adjacent to cellular nuclear matrix-associated ND10 sites. During productive infection, the HSV single-stranded DNA-binding protein ICP8 localizes to replication compartments. To further the understanding of replication compartment maturation, we have constructed and characterized a recombinant HSV-1 strain that expresses an ICP8 molecule with green fluorescent protein (GFP) fused to its C terminus. In transfected Vero cells that were infected with HSV, the ICP8-GFP protein localized to prereplicative sites in the presence of the viral DNA synthesis inhibitor phosphonoacetic acid (PAA) or to replication compartments in the absence of PAA. A recombinant HSV-1 strain expressing the ICP8-GFP virus replicated in Vero cells, but the yield was increased by 150-fold in an ICP8 complementing cell line. Using the ICP8-GFP protein as a marker for replication compartments, we show here that these structures start as punctate structures early in infection and grow into large, globular structures that eventually fill the nucleus. Large replication compartments were formed by small structures that either moved through the nucleus to merge with adjacent compartments or remained relatively stationary within the nucleus and grew by accretion and fused with neighboring structures. PMID- 12758172 TI - Screening for simian foamy virus infection by using a combined antigen Western blot assay: evidence for a wide distribution among Old World primates and identification of four new divergent viruses. AB - Simian foamy viruses (SFVs) belong to a genetically and antigenically diverse class of retroviruses that naturally infect a wide range of nonhuman primates (NHPs) and can also be transmitted to humans occupationally exposed to NHPs. Current serologic detection of SFV infection requires separate Western blot (WB) testing by using two different SFV antigens [SFV(AGM) (African green monkey) and SFV(CPZ) (chimpanzee)]. However, this method is labor intensive and validation is limited to only small numbers of NHPs. To facilitate serologic SFV testing, we developed a WB assay that combines antigens from both SFV(AGM) and SFV(CPZ). The combined-antigen WB (CA-WB) assay was validated with 145 serum samples from 129 NHPs (32 African and Asian species) and 16 humans, all with known SFV infection status determined by PCR. Concordant CA-WB results were obtained for all 145 PCR positive or -negative primate and human specimens, giving the assay a 100% sensitivity and specificity. In addition, no reactivity was observed in sera from persons positive for human immunodeficiency virus or human T cell lymphotropic virus (HIV/HTLV) (n = 25) or HIV/HTLV-negative U.S. blood donors (n = 100). Using the CA-WB assay, we screened 360 sera from 43 Old World primate species and found an SFV prevalence of about 68% in both African and Asian primates. We also isolated SFV from the blood of four seropositive primates (Allenopithecus nigroviridis, Trachypithecus francoisi, Hylobates pileatus, and H. leucogenys) not previously known to be infected with SFV. Phylogenetic analysis of integrase sequences from these isolates confirmed that all four SFVs represent new, distinct, and highly divergent lineages. These results demonstrate the ability of the CA-WB assay to detect infection in a large number of NHP species, including previously uncharacterized infections with divergent SFVs. PMID- 12758173 TI - Analysis of intermolecular RNA-RNA recombination by rubella virus. AB - To investigate whether rubella virus (RUB) undergoes intermolecular RNA-RNA recombination, cells were cotransfected with pairs of in vitro transcripts from genomic cDNA plasmid vectors engineered to contain nonoverlapping deletions: the replicative transcript maintained the 5'-proximal nonstructural (NS) ORF (which contained the replicase, making it RNA replication competent), had a deletion in the 3'-proximal structural protein (SP) ORF, and maintained the 3' end of the genome, including the putative 3' cis-acting elements (CSE), while the nonreplicative transcript consisted of the 3' half of the genome including the SP ORF and 3' CSE. Cotransfection yielded plaque-forming virus that synthesized the standard genomic and subgenomic RNAs and thus was generated by RNA-RNA recombination. Using transcripts tagged with a 3'-terminal deletion, it was found that recombinants contained the 3' end derived from the replicative strand, indicating a cis-preference for initiation of negative-strand synthesis. In cotransfections in which the replicative transcript lacked the 3' CSE, recombination occurred, albeit at lower efficiency, indicating that initiation in trans from the NS-ORF can occur. The 3' CSE was sufficient as a nonreplicative transcript, showing that it can serve as a promoter for negative-strand RNA synthesis. While deletion mutagenesis showed that the presence of the junction untranslated region (J-UTR) between the ORFs appeared to be necessary on both transcripts for recombination in this region of the genome, analysis with transcripts tagged with restriction sites showed that the J-UTR was not a hot spot for recombination compared to neighboring regions in both ORFs. Sequence analysis of recombinants revealed that both precise (homologous) and imprecise recombination (aberrant, homologous resulting in duplications) occurred; however, imprecise recombination only involved the J-UTR or the 3' end of the NS-ORF and the J-UTR (maintaining the NS-ORF), indicating selection pressure against duplications in other regions of the genome. PMID- 12758174 TI - Enhanced cellular immune response against SIV Gag induced by immunization with DNA vaccines expressing assembly and release-defective SIV Gag proteins. AB - Codon-optimized genes were synthesized for the SIVmac239 Gag, a mutant Gag with mutations in the major homology region, and a chimeric Gag containing a protein destruction signal at the N-terminus of Gag. The mutant and chimeric Gag were expressed at levels comparable to that observed for the wild-type Gag protein but their stability and release into the medium were found to be significantly reduced. Immunization of mice with DNA vectors encoding the mutant or chimeric Gag induced fourfold higher levels of anti-SIV Gag CD4 T cell responses than the DNA vector encoding the wild-type SIV Gag. Moreover, anti-SIV Gag CD8 T cell responses induced by DNA vectors encoding the mutant or chimeric Gag were found to be 5- to 10-fold higher than those induced by the DNA construct for the wild type Gag. These results indicate that mutations disrupting assembly and/or stability of the SIV Gag protein effectively enhance its immunogenicity when expressed from DNA vaccines. PMID- 12758175 TI - A mosaic adenovirus possessing serotype Ad5 and serotype Ad3 knobs exhibits expanded tropism. AB - The efficiency of cancer gene therapy with recombinant adenoviruses based on serotype 5 (Ad5) has been limited partly because of variable, and often low, expression by human primary cancer cells of the primary cellular-receptor which recognizes the knob domain of the fiber protein, the coxsackie and adenovirus receptor (CAR). As a means of circumventing CAR deficiency, Ad vectors have been retargeted by utilizing chimeric fibers possessing knob domains of alternate Ad serotypes. We have reported that ovarian cancer cells possess a primary receptor for Ad3 to which the Ad3 knob binds independently of the CAR-Ad5 knob interaction. Furthermore, an Ad5-based chimeric vector, designated Ad5/3, containing a chimeric fiber proteins possessing the Ad3 knob, demonstrates CAR independent tropism by virtue of targeting the Ad3 receptor. Based on these findings, we hypothesized that a mosaic virus possessing both the Ad5 knob and the Ad3 knob on the same virion could utilize either primary receptor, resulting in expanded tropism. In this study, we generated a dual-knob mosaic virus by coinfection of 293 cells with Ad5-based and Ad5/3-based vectors. Characterization of the resultant virions confirmed the incorporation of both Ad5 and Ad3 knobs in the same particle. Furthermore, this mosaic virus was able to utilize either receptor, CAR and the Ad3 receptor, for virus attachment to cells. Enhanced Ad infectivity with the mosaic virus was shown in a panel of cell lines, with receptor profiles ranging from CAR-dominant to Ad3 receptor-dominant. Thus, this mosaic virus strategy may offer the potential to improve Ad-based gene therapy approaches by infectivity enhancement and tropism expansion. PMID- 12758176 TI - Mutations that abrogate transactivational activity of the feline leukemia virus long terminal repeat do not affect virus replication. AB - The U3 region of the LTR of oncogenic Moloney murine leukemia virus (Mo-MuLV) and feline leukemia viruses (FeLV) have been previously reported to activate expression of specific cellular genes in trans, such as MHC class I, collagenase IV, and MCP-1, in an integration-independent manner. It has been suggested that transactivation of these specific cellular genes by leukemia virus U3-LTR may contribute to the multistage process of leukemogenesis. The U3-LTR region, necessary for gene transactivational activity, also contains multiple transcription factor-binding sites that are essential for normal virus replication. To dissect the promoter activity and the gene transactivational activity of the U3-LTR, we conducted mutational analysis of the U3-LTR region of FeLV-A molecular clone 61E. We identified minimal nucleotide substitution mutants on the U3 LTR that did not disturb transcription factor-binding sites but abrogated its ability to transactivate the collagenase gene promoter. To determine if these mutations actually have altered any uncharacterized important transcription factor-binding site, we introduced these U3-LTR mutations into the full-length infectious molecular clone 61E. We demonstrate that the mutant virus was replication competent but could not transactivate cellular gene expression. These results thus suggest that the gene transactivational activity is a distinct property of the LTR and possibly not related to its promoter activity. The cellular gene transactivational activity-deficient mutant FeLV generated in this study may also serve as a valuable reagent for testing the biological significance of LTR-mediated cellular gene activation in the tumorigenesis caused by leukemia viruses. PMID- 12758177 TI - Signals in hepatitis A virus P3 region proteins recognized by the ubiquitin mediated proteolytic system. AB - The hepatitis A virus 3C protease and 3D RNA polymerase are present in low concentrations in infected cells. The 3C protease was previously shown to be rapidly degraded by the ubiquitin/26S proteasome system and we present evidence here that the 3D polymerase is also subject to ubiquitination-mediated proteolysis. Our results show that the sequence (32)LGVKDDWLLV(41) in the 3C protease serves as a protein destruction signal recognized by the ubiquitin protein ligase E3alpha and that the destruction signal for the RNA polymerase does not require the carboxyl-terminal 137 amino acids. Both the viral 3ABCD polyprotein and the 3CD diprotein were also found to be substrates for ubiquitin mediated proteolysis. Attempts to determine if the 3C protease or the 3D polymerase destruction signals trigger the ubiquitination and degradation of these precursors yielded evidence suggesting, but not unequivocally proving, that the recognition of the 3D polymerase by the ubiquitin system is responsible. PMID- 12758178 TI - The immunodominant influenza matrix T cell epitope recognized in human induces influenza protection in HLA-A2/K(b) transgenic mice. AB - The protective efficacy of the influenza matrix protein epitope 58-66 (called M1), recognized in the context of human HLA-A2 molecules, was evaluated in a HLA A2/K(b) transgenic mouse model of lethal influenza infection. Repeated subcutaneous immunizations with M1 increased the percentage of survival. This effect was mediated by T cells since protection was abolished following in vivo depletion of all T lymphocytes, CD8(+), or CD4(+) T cells. The survival correlated with the detection of memory CD8(+) splenocytes able to proliferate in vitro upon stimulation with M1 and to bind M1-loaded HLA-A2 dimers, as well as with M1-specific T cells in the lungs, which were directly cytotoxic to influenza infected cells following influenza challenge. These results demonstrated for the first time that HLA-A2-restricted cytotoxic T cells specific for the major immunodominant influenza matrix epitope are protective against the infection. They encourage further in vivo evaluation of T cell epitopes recognized in the context of human MHC molecules. PMID- 12758179 TI - Cre/loxP-mediated adenovirus type 5 packaging signal excision demonstrates that core element VI is sufficient for virus packaging. AB - Previous analyses have demonstrated that packaging of the adenovirus type 5 (Ad5) genome is dependent on at least seven cis-acting elements, called AI to AVII, which are located in the left-end region of the genome. These elements have different packaging efficiencies, and without AI through AV, viral DNA cannot be packaged. Here we report the identification of the cis-acting Ad5 packaging domain in vivo by using the Cre/loxP system. We found that an adenoviral DNA fragment (nt 192 to nt 358), which includes elements AI to AV, is excised by Cre recombinase and packaged into capsids. Furthermore, this mutant adenovirus replicated so efficiently by repetitive propagation that its purification by CsCI equilibrium gradient was possible. This study clarified that the region from nt 358 to nt 454 on the viral genome is sufficient for packaging. Recently, the helper-dependent adenoviral vector (HDAd) construction system has been developed for the purpose of gene therapy. This system uses a helper virus with two parallel loxP sites flanking the packaging signal. This region is eliminated by Cre-mediated excision, which prevents helper virus packaging. Our data provide useful information regarding factors affecting efficient elimination. PMID- 12758180 TI - A long HBV transcript encoding pX is inefficiently exported from the nucleus. AB - The longest hepatitis B virus transcript is a 3.9-kb mRNA whose function remained unclear. In this study, we wished to identify the translation products and physiological role of this viral transcript. This transcript initiates from the X promoter region ignoring the inefficient and noncanonical viral polyadenylation signal at the first round of transcription. However, an HBV mutant with canonical polyadenylation signal continues, though with lower efficiency, to program the synthesis of this long transcript, indicating that the deviated HBV polyadenylation signal is important but not essential to enable transcription of the 3.9-kb species. The 3.9-kb RNA contains two times the X open reading frame (ORF). The X ORF at the 5'-end is positioned upstream of the CORE gene. By generating an HBV DNA mutant in which the X and Core ORFs are fused, we demonstrated the production of a 40-kDa X-Core fusion protein that must be encoded by the 3.9-kb transcript. Mutagenesis studies revealed that the production of this protein depends on the 5' X ORF ATG, suggesting that the 3.9 kb RNA is active in translation of the X ORF. Based on these features, the 3.9-kb transcript was designated lxRNA for long X RNA. Unlike other HBV transcripts, lxRNA harbors two copies of PRE, the posttranscriptional regulatory element that controls the nuclear export of HBV mRNAs. Unexpectedly, despite the presence of PRE sequences, RNA fractionation analysis revealed that lxRNA barely accumulates in the cytoplasm, suggesting that nuclear export of lxRNA is poor. Collectively, our data suggest that two distinct HBV mRNA species encode pX and that the HBV transcripts are differentially regulated at the level of nuclear export. PMID- 12758181 TI - The Fourth World Congress on Tuberculosis. PMID- 12758183 TI - The TB epidemic from 1992 to 2002. AB - In 1992, less than 20 countries were implementing a sound TB control strategy. At the same time, TB was being resurrected as a major public health problem world wide after two decades of neglect. Awareness of upward trends in the industrialized countries and MDR-TB outbreaks in large cities were driving forces behind the re-emergence of TB in the international health agenda. New evidence, and consequent estimates, suggested that the situation in developing countries, especially in sub-Saharan Africa, was deteriorating rapidly. Similarly, major increases were observed in the former USSR. It was estimated that some 7-8 million new cases and 2-3 million deaths were occurring annually in the world. The global targets of reaching 85% cure rates and 70% case detection among infectious cases were established by the World Health Assembly in 1991. Both the WHO declaration of TB as a global emergency in 1993 and the launch of the five element DOTS strategy in 1994-1995 resulted in countries adopting DOTS in encouraging numbers. In fact, in 2000, 148 countries including all 22 highest burden countries (HBC) responsible for 80% of cases world-wide, had adopted the new DOTS strategy. Nevertheless, progress in case detection remained slow due to incomplete geographical coverage or need to widen detection and notification capacity with innovative schemes. The major constraints to TB control became increasingly clear, and a global Stop TB Partnership was eventually established to address such constraints. A Global DOTS Expansion Plan revealed the needs and the gaps to achieve the global targets in 2005. Today, in 2002, the top priority remains that of expanding DOTS, as rapidly as possible, using a number of new approaches to increase case detection and notification while maintaining high cure rates. These must involve collaboration with the private sector and communities, as well as strengthening of primary care services. Similarly, crucial is the rapid identification of solutions to TB/HIV and MDR-TB. PMID- 12758184 TI - Tuberculosis control in China. PMID- 12758185 TI - Tuberculosis trends in the United States, 1992-2001. AB - At the time of the last world congress on tuberculosis (TB) in 1992, the United States (US) was experiencing an unprecedented resurgence of TB. Since the mid 1950s, TB incidence had been steadily decreasing, until 1984 when this longstanding trend was reversed. The annual national total of TB cases continued to increase and peaked in 1992 with 26,673 TB cases reported (10.5 TB cases per 100,000 population). A prompt and formidable response from local, state, and federal governments helped curb the resurgence. From 1992 to 2001, total TB incidence decreased by 40% to an all-time low of 15,989 TB cases reported in 2001. The decrease in TB cases from 2000 to 2001, however, was the smallest (2.4%) since the resurgence a decade ago. This report will briefly review the trends and factors associated with the TB resurgence in the late 1980s and early 1990s, and provide a detailed description of specific TB trends in the US between 1992 and 2001. PMID- 12758186 TI - DOTS progress in India: 1995-2002. PMID- 12758187 TI - What is the limit to case detection under the DOTS strategy for tuberculosis control? AB - In year 2000, the WHO DOTS strategy for tuberculosis (TB) control had been adopted by 148 out of 212 countries, but only 27% of all estimated sputum smear positive patients were notified under DOTS in that year. Here we investigate the way in which gains in case detection under DOTS were made up until 2000 in an attempt to anticipate future progress towards the global target of 70% case detection. The analysis draws on annual reports of DOTS geographical coverage and case notifications, and focuses on the 22 high-burden countries (HBCs) that account for about 80% of new TB cases arising globally each year. Our principal observation is that, as TB programmes in the 22 HBCs have expanded geographically, the fraction of the estimated number of sputum smear-positive cases detected within designated DOTS areas has remained constant at 40-50% although there are significant differences between countries. This fraction is about the same as the percentage of all smear-positive cases notified annually to WHO via public health systems worldwide. The implication is that, unless the DOTS strategy can reach beyond traditional public health reporting systems, or unless these systems can be improved, case detection will not rise much above 40% in the 22 HBCs, or in the world as a whole, even when the geographical coverage of DOTS is nominally 100%. We estimate that, under full DOTS coverage, three-quarters of the undetected smear-positive cases will be living in India, China, Indonesia, Nigeria, Bangladesh and Pakistan. But case detection could also remain low in countries with smaller populations: in year 2000, over half of all smear-positive TB cases were living in 49 countries that detected less than 40% of cases within DOTS areas. Substantial efforts are therefore needed (a) to develop new case finding and management methods to bridge the gap between current and target case detection, and (b) to improve the accuracy of national estimates of TB incidence, above all by reinforcing and expanding routine surveillance. PMID- 12758188 TI - The global situation of MDR-TB. AB - Drug-resistant tuberculosis has been reported since the early days of the introduction of chemotherapy. However, most of the evidence was limited to developed countries. In 1992, the Third World Congress on Tuberculosis concluded that there was little recent information on the global magnitude of multidrug resistant tuberculosis (MDR-TB), defined as resistance to at least isoniazid and rifampicin. Through the WHO/IUATLD Global Project on Drug-Resistance Surveillance launched in 1994, a large number of reliable and accurate data have allowed us to understand the magnitude of the problem of MDR-TB. The data available suggest that globally MDR-TB is not a problem (median = 1% in 64 countries/geographical sites surveyed) of the same magnitude as that of drug-susceptible tuberculosis. However, MDR-TB is at critical levels in specific regions of the world. Hot spots for MDR-TB include Estonia, Latvia, the Oblasts of Ivanovo and Tomsk in Russia, and the provinces of Henan and Zhejiang Provinces in China. Trends confirm that MDR-TB is limited to local epidemics but the evidence is not yet irrefutable, as many countries have only provided short-term data. Two-thirds of the world's countries and, more importantly, half of the 22 tuberculosis high-burden countries, have not yet provided data. Mathematical modelling suggests that 3.2% (or 273,000) of the world's estimated new tuberculosis cases (95% confidence intervals: 185,000 and 414,000) were MDR-TB in 2000. Adoption of DOTS to prevent the generation of resistant strains and careful introduction of second-line drugs to treat patients with MDR are the top priorities for proper control/containment of MDR-TB. PMID- 12758189 TI - DOTS-Plus for multidrug-resistant tuberculosis in the Philippines: global assistance urgently needed. AB - SETTING: The Philippines, a high burden country for tuberculosis (TB). STUDY DESIGN: Health Operational Study. OBJECTIVE: To describe preliminary data from the Makati Medical Center (MMC)-DOTS Plus pilot project. METHODS: Patients were consecutively enrolled after confirmation of MDR-TB status. Individualized treatment regimens were based on drug susceptibility testing and history of previous intake for the other drugs that were not tested. Treatment outcome in those who had completed at least 18 months of therapy and interim outcome for those who received more than 12 months but less than 18 months were analyzed. RESULTS: One hundred forty-nine patients with MDR-TB were enrolled from April 1999 to 30 May 2002 at the MMC DOTS Clinic. Referrals were from private institutions and practicing physicians in 73.2% of cases. Approximately 30% of isolates tested were resistant to all five first-line drugs, 39.4% to four, 16.8% to three, 12.1% to two. Fluoroquinolone resistance was noted in 40.9% of all the isolates, including 54.5% of those resistant to five drugs and 34.6% of those resistant to four drugs. The outcome of 23 patients who completed therapy and 62 who have received more than 12 months therapy showed cure and likely cure in 73.4% of cases and failure in 3.8% and likely failure in 6.3%. Death occurred in 3.8% and default was observed in 11.4%. CONCLUSION: The MMC DOTS-Plus pilot project is a public-private collaboration in TB Control. Response to therapy was encouraging. Complete subsidy of medicines and laboratory and clinic services and DOT were essential in the successful implementation of the program. DOTS-Plus and DOTS should go hand in hand in TB control if MDR-TB is highly prevalent. PMID- 12758190 TI - From multidrug-resistant tuberculosis to DOTS expansion and beyond: making the most of a paradigm shift. AB - This review examines the paradigm shift around multidrug-resistant tuberculosis (MDR-TB). This shift has centered largely on the activities of institutions participating in the World Health Organization's Working Group on DOTS-Plus for MDR-TB. We review the important milestones since 1995, namely, the emergence of new evidence, the construction of new mechanisms, and the building of consensus to support new policy guidelines. This paper offers a case study of the construction of a model of good global governance that--if the opportunity is taken--can improve access to effective TB care through improving and helping to expand the WHO-recommended DOTS strategy. PMID- 12758191 TI - The impact of HIV on transmission of tuberculosis in Tanzania. PMID- 12758192 TI - Can we control tuberculosis in high HIV prevalence settings? AB - The overlap between the epidemiology of HIV and tuberculosis and consequent rapid rise in numbers of patients with tuberculosis in many African countries has put a huge burden on health systems. The stigma of HIV has increased the existing stigma surrounding tuberculosis. There are three mechanisms by which we may reduce the number of cases of tuberculosis in a community: reducing transmission of tuberculosis, reducing reactivation of latent tuberculosis infection and reducing HIV transmission. Reinforcing the existing health service to find more cases, active case-finding in communities or enhanced case-finding in specific groups will reduce transmission of tuberculosis. However, health services that find it difficult to find cases efficiently will also find it difficult to support patients throughout treatment to achieve a cure. Partnership with traditional healers, community-based organizations and private practitioners could reduce this burden. Reactivation of tuberculosis among people living with HIV can be reduced by tuberculosis preventive therapy or by antiretroviral therapy. Programmes that identify people living with HIV can also implement enhanced tuberculosis case-finding increasing the benefits of the programme. However, the impact of widespread use of antiretroviral therapy may be to increase the number of people in a community who are mildly immunocompromised and the incidence of tuberculosis at a community level might rise. Any strategy that successfully reduces HIV transmission will benefit tuberculosis control, since around a third of all HIV-positive individuals will develop tuberculosis before they die. To control tuberculosis in high HIV prevalence settings, we must strengthen health systems to include not only expansion of the DOTS strategy but also full-blooded implementation of voluntary counselling and testing, enhanced and active tuberculosis case-finding, preventive therapy and better care for people living with HIV including antiretroviral therapy. The approach needed to control tuberculosis needs also to be integrated into broader development and poverty reduction goals. PMID- 12758193 TI - Ten years of research progress and what's to come. AB - There has been a renaissance in interest in tuberculosis research over the last decade. A search of the National Library of Medicine database records an output of 246 papers on Mycobacterium tuberculosis in 1980. This had risen to 615 in 1990, to over 1000 in 1995, and to 1537 in the year 2000. This increase has been stimulated by heightened awareness amongst the research community of the magnitude of the global burden of tuberculosis, by increased funding, and by new scientific opportunities provided by advances in genomics and in cellular immunology. PMID- 12758194 TI - Tuberculosis control: past 10 years and future progress. AB - The number of countries implementing directly observed therapy short-course (DOTS) has grown rapidly in the past decade and more than 10 million patients have now been treated under DOTS. While global case detection rates increased slightly, from 35% to 40% between 1995 and 2000, the proportion attributable to DOTS grew from less than one-third to more than two-thirds. DOTS is replacing inferior treatment but still treating fewer than 40% of estimated new TB cases. Misconceptions threaten to undermine continued success in tuberculosis control. The first misconception is that treatment observation is unnecessary. Treatment observation needs to be made more patient-friendly, but must not be abandoned. The second misconception is that health care reform will strengthen tuberculosis control. TB control is essentially a management problem. Greater accountability of governments, donors and providers is essential. A third misconception is to focus on treating multi-drug-resistant tuberculosis (MDRTB) cases without addressing the root causes of MDRTB. While it is important, on a clinical basis and epidemiologically in some contexts, to care optimally for patients with MDRTB, it is more important to address the cause of MDRTB and to fix the program generating MDRTB. The fourth misconception is an inordinate concern for sustainability. Delaying assistance will make implementation and sustainability in the future more difficult. Tuberculosis control is remarkably inexpensive and cost-effective, but efforts will fail unless programs have the ability to hire staff, purchase supplies, and contract for services efficiently. Critical issues for the future of tuberculosis control are sustained funding, technical rigor, and good management. PMID- 12758195 TI - A short history of Robert Koch's fight against tuberculosis: those who do not remember the past are condemned to repeat it. PMID- 12758196 TI - Structure, function, and biogenesis of the cell wall of Mycobacterium tuberculosis. AB - Much of the early structural definition of the cell wall of Mycobacterium spp. was initiated in the 1960s and 1970s. There was a long period of inactivity, but more recent developments in NMR and mass spectral analysis and definition of the M. tuberculosis genome have resulted in a thorough understanding, not only of the structure of the mycobacterial cell wall and its lipids but also the basic genetics and biosynthesis. Our understanding nowadays of cell-wall architecture amounts to a massive "core" comprised of peptidoglycan covalently attached via a linker unit (L-Rha-D-GlcNAc-P) to a linear galactofuran, in turn attached to several strands of a highly branched arabinofuran, in turn attached to mycolic acids. The mycolic acids are oriented perpendicular to the plane of the membrane and provide a truly special lipid barrier responsible for many of the physiological and disease-inducing aspects of M. tuberculosis. Intercalated within this lipid environment are the lipids that have intrigued researchers for over five decades: the phthiocerol dimycocerosate, cord factor/dimycolyltrehalose, the sulfolipids, the phosphatidylinositol mannosides, etc. Knowledge of their roles in "signaling" events, in pathogenesis, and in the immune response is now emerging, sometimes piecemeal and sometimes in an organized fashion. Some of the more intriguing observations are those demonstrating that mycolic acids are recognized by CD1-restricted T-cells, that antigen 85, one of the most powerful protective antigens of M. tuberculosis, is a mycolyltransferase, and that lipoarabinomannan (LAM), when "capped" with short mannose oligosaccharides, is involved in phagocytosis of M. tuberculosis. Definition of the genome of M. tuberculosis has greatly aided efforts to define the biosynthetic pathways for all of these exotic molecules: the mycolic acids, the mycocerosates, phthiocerol, LAM, and the polyprenyl phosphates. For example, we know that synthesis of the entire core is initiated on a decaprenyl-P with synthesis of the linker unit, and then there is concomitant extension of the galactan and arabinan chains while this intermediate is transported through the cytoplasmic membrane. The final steps in these events, the attachment of mycolic acids and ligation to peptidoglycan, await definition and will prove to be excellent targets for a new generation of anti-tuberculosis drugs. PMID- 12758197 TI - Human immunity to M. tuberculosis: T cell subsets and antigen processing. AB - A hallmark of M. tuberculosis infection is the ability of most (90-95%) healthy adults to control infection through acquired immunity, in which antigen specific T cells and macrophages arrest growth of M. tuberculosis bacilli and maintain control over persistent bacilli. In addition to CD4+ T cells, other T cell subsets such as, gammadelta, CD8+ and CD1-restricted T cells have roles in the immune response to M. tuberculosis. A diverse T cell response allows the host to recognize a wider range of mycobacterial antigens presented by different families of antigen-presenting molecules, and thus greater ability to detect the pathogen. Macrophages are key antigen presenting cells for T cells, and M. tuberculosis survives and persists in this central immune cell. This is likely an important factor in generating this T cell diversity. Furthermore, the slow growth and chronic nature of M. tuberculosis infection results in prolonged exposure to antigens, and hence further T cell sensitization. The effector mechanisms used by T cells to control M. tuberculosis are poorly understood. To survive in macrophages, M. tuberculosis has evolved mechanisms to block immune responses. These include modulation of phagosomes, neutralization of macrophage effector molecules, stimulating the secretion of inhibitory cytokines, and interfering with processing of antigens for T cells. The relative importance of these blocking mechanisms likely depends on the stage of M. tuberculosis infection: primary infection, persistence, reactivation or active tuberculosis. The balance of the host-pathogen interaction in M. tuberculosis infection is determined by the interaction of T cells and infected macrophages. The outcome of this interaction results either in control of M. tuberculosis infection or active disease. A better understanding of this interaction will result in improved approaches to treatment and prevention of tuberculosis. PMID- 12758198 TI - Immune response to tuberculosis: experimental animal models. PMID- 12758199 TI - The mouse as a useful model of tuberculosis. AB - Like other animal models of tuberculosis, the mouse has provided a large amount of information that can be applied to understanding the disease process in infected humans. The model is particularly useful in providing information about the immune response, given the huge database of reagents now available, including antibodies to lymphocyte markers and the growing number of available gene disrupted mice, and the model is validated by the fact that multiple mechanisms discovered in the mouse such as the TH1 pathway and the Toll-like receptor system are similarly important in humans. The model also has its limitations, particularly in terms of the immunopathologic response, in which similar elements occur but are expressed somewhat differently. PMID- 12758200 TI - Non-human primates: a model for tuberculosis research. AB - A variety of animals have been used for tuberculosis research, and each animal model has its strengths and weaknesses. We sought to develop a non-human primate model of tuberculosis to model aspects of human tuberculosis that are difficult to model in other animals, including the pathology in the lungs, various progression to disease, and immunologic correlates of infection or disease that are likely to be similar in humans. To date, we have infected 17 cynomolgus macaques (Macaca fasicularis) with a low dose (15-25CFU) of Mycobacterium tuberculosis strain Erdman. The monkeys were grouped into three categories on the basis of disease progression: rapid progression (advanced disease by 3 months post-infection), active/chronic infection (signs of disease but a slower progression), and latent infection (no signs of clinical disease). Animals were followed clinically post-infection, including blood work, physical examinations, serial bronchoalveolar lavage (BAL) and gastric aspirates for M. tuberculosis culture, chest radiographs, and tuberculin reactivity. Immunologic assays on cells from blood, BAL fluid, and tissue, have been performed, including proliferation, flow cytometry, ELIspot assays, cytotoxic T lymphocyte (CTL) assays, and ELISAs. The spectrum of disease observed in these monkeys is similar to humans, and this model may be very useful for studying pathogenesis and immunology of tuberculosis, as well as testing vaccines, diagnostic reagents, and drugs prior to use in human populations. PMID- 12758201 TI - Use of the bovine model of tuberculosis for the development of improved vaccines and diagnostics. AB - Over the past few years there has been a resurgence in research into bovine tuberculosis due to the sharp rise of the disease in countries such as Great Britain and to the continuing problem of wild-life reservoirs in countries such as New Zealand. One of the goals of this research is to develop cattle vaccines against TB. The initial testing of candidate vaccines is carried out in laboratory animals, initially mice and subsequently guinea pigs. A unique feature of the cattle vaccination programme is that candidate vaccines which show promise in laboratory models can then be tested in the natural host species, cattle, before progressing to clinical trials. This is a major advantage over the strategy for developing a vaccine for human tuberculosis where, of course, it is impossible to test a candidate vaccine by experimentally challenging the host species with the pathogen. The most commonly used model for testing vaccine candidates in cattle consists of an intra-tracheal challenge of between 10(3) and 10(4) colony forming units of Mycobacterium bovis. The pathology observed following challenge is similar to human tuberculosis giving rise to a marked granulomatous reaction and a predominantly cellular immune response. Using this model we have been able to make a number of significant advances towards a bovine TB vaccine. First we have developed antigen cocktails that, when used in a whole blood gamma interferon assay, can differentiate between M. bovis infected and BCG vaccinated animals. Next we have developed immune correlates of pathology, which allow us to assess whether the vaccine is protecting animals against challenge before post mortem examination. Finally we have been able to use the model to develop a vaccine that improves the efficacy of BCG against M. bovis challenge. PMID- 12758202 TI - Hematogenous reseeding of the lung in low-dose, aerosol-infected guinea pigs: unique features of the host-pathogen interface in secondary tubercles. AB - The ability to study the early events in the pathogenesis of pulmonary tuberculosis in guinea pigs following very low dose (3-5 cfu) infection by the respiratory route has revealed that early (10-14 days) extrapulmonary dissemination results in reseeding of previously uninfected lobes of the lung by the hematogenous route. Thus, in every guinea pig, the lung is challenged twice, once by the airway and 2-3 weeks later by the circulatory system. The so called "secondary" pulmonary lesions which result from the bacillemia differ fundamentally from the primary lesions, in part, because the host has already developed a strong T cell mediated immunity when the hematogenous reseeding occurs. Secondary lung lesions in non-vaccinated guinea pigs behave similarly to primary lung lesions in previously vaccinated guinea pigs. Since the secondary, blood-borne lesions are thought to be the "reactivatable foci" which result in reactivation tuberculosis following prolonged persistent infection, it is important to understand the nature of the host-pathogen interaction in secondary lesions. The guinea pig model provides a unique opportunity to examine both the microbial and host factors which constitute that interface. PMID- 12758203 TI - Shifting the focus of tuberculosis research in India. AB - India has a long and distinguished tradition of research in the field of tuberculosis (TB). Pioneering studies from India demonstrated the efficacy and safety of domiciliary treatment, the necessity of direct observation of treatment, the feasibility of case detection through sputum smear microscopy in primary health care institutions, and the effectiveness of intermittent short course chemotherapy. These findings laid the foundation of directly observed treatment, short course (DOTS), which has been adopted by nearly 150 countries worldwide. Today, India has the second-largest and the fastest-growing DOTS programme in the world. A strong component of programme evaluation and operational research is needed to sustain and expand DOTS in the context of a suboptimal primary health care system, a large and unregulated private health care system, and the dual threats of HIV and multidrug-resistant TB (MDR-TB). Therefore, the focus of TB research in India has shifted to the following operational research areas: evaluating models to involve the private health sector; assessing the role of incentives in increasing treatment compliance; examining gender differentials in the access to TB services; assessing risk factors for delay in diagnosis; evaluating diagnosis, treatment and prevention of TB among HIV-infected persons; monitoring MDR-TB; estimating cost-effectiveness of the DOTS programme; monitoring the quality of smear microscopy services; and measuring the current burden of TB. Research for developing newer diagnostic tools, drugs and vaccines remains a long-term priority. Greater networking is needed among national researchers, programme managers and policy-makers to translate the findings of research into policies and programmes to make TB control in India more effective and efficient. PMID- 12758204 TI - Integration of operational research into National Tuberculosis Control Programmes. AB - Operational research, within the context of a national disease control programme, may be described as the search for knowledge on interventions, tools or strategies which enhance programme effectiveness. There are two examples from Malawi of how operational research into recurrent tuberculosis and decentralization of treatment lead to information which enabled the National TB Control Programme (NTP) to change and improve its practice and policy. The key factors which allowed this to happen, and the guiding principles about integrating research into national programmes are discussed. TB programmes must have clear objectives, be able to identify constraints which prevent objectives being met and ask research questions around these constraints. There must be sufficient resources for research, both material and financial, and this requires programmes to incorporate a research agenda into their costed annual workplans. Training is also a key component of developing an integrated research programme, and should be included in the budgets. Research outputs should be judged in terms of activities undertaken and completed, papers written, and regular documentation of how research has influenced policy and practice. Finally, there should be strong advocacy for operational research, so that government policy makers can be convinced of its value. PMID- 12758205 TI - Involving private practitioners and chest physicians in the control of tuberculosis. AB - SETTING: Private practitioners including chest physicians are often an important obstacle to having a successful National Tuberculosis Programme (NTP) in low- and middle-income countries. This complicated situation has been present in most of the Latin American countries for many years. OBJECTIVE: To design an intervention model to obtain collaboration and integration of specialist physicians in the actions of the NTP. DESIGN: In 1998, the IUATLD designed a special interactive model of training courses, to be held in an important number of Latin American countries. This intensive (25h in 3 days) course was named "Importance of the role of chest physicians and their integration in NTP strategies". At the end of each course, the participants were invited to sign a series of agreements concerning controversial topics that had blocked collaboration in the past. RESULTS: This course, adapted to the situation of the different countries, has been held 17 times in 9 different countries. So far, nearly 600 specialist physicians have been trained with this special model, and all of them have signed important agreements on future collaboration. CONCLUSION: There has been an important improvement in integrating these specialist physicians into the actions of the NTPs. This intervention has contributed to a substantial improvement of tuberculosis control in the last 4 years in Latin America. PMID- 12758206 TI - Involving private health care providers in delivery of TB care: global strategy. AB - Most poor countries have a large and growing private medical sector. Evidence suggests that a large proportion of tuberculosis patients in many high TB- burden countries first approach a private health care provider. Further, private providers manage a significant proportion of tuberculosis cases. Surprisingly though, there is virtually no published evidence on linking private providers to tuberculosis programmes. As a part of global efforts to control tuberculosis through effective DOTS implementation, the World Health Organization has recently begun addressing the issue of private providers in TB control through an evolving global strategy. As a first step, a global assessment of private providers' participation in tuberculosis programmes was undertaken. The findings of the assessment were discussed and debated in a consultation involving private practitioners, TB programme managers and policy makers. Their recommendations have contributed to the evolving global strategy called Public-Private Mix for DOTS implementation (PPM DOTS). This paper presents the guiding principles of PPM DOTS and major elements of the global strategy. These include: informed advocacy; setting-up "learning projects"; scaling-up successful projects and formulation of regional, national and local strategies; developing practical tools to facilitate PPM DOTS and pursuing an operational research agenda to help better design and shape PPM DOTS strategies. Encouraging results from some ongoing project sites are discussed. The paper concludes that concerted global efforts and local input are required for a sustained period to help achieve productive engagement of private practitioners in DOTS implementation. Such efforts have to be targeted as much towards national tuberculosis programmes as towards private providers and their associations. Continued apathy in this area could not only potentially delay achieving global targets for TB control but also undo, in the long run, the hard-earned achievements of National TB Programmes. PMID- 12758207 TI - The Mumbai experience in building field level partnerships for DOTS implementation. AB - In February 1999, the Revised National Tuberculosis (TB) Control Programme (RNTCP) was implemented in the city of Mumbai after a pilot phase of 5 years. The city has a population of more than 12 million people and an estimated annual TB incidence of 21,000 cases, 8000 of these being infectious. This paper describes a partnership between the TB programme and a Non Governmental Organization (NGO), which began with a methodological analysis of the problems faced by the programme to help identify other key organizations, who might usefully be involved. The work focussed on "networking" to ensure the optimum use of existing resources. The problems encountered affected all levels of TB control from access to drug supply and treatment. The major issues related to an inadequate public health infrastructure resulting in poor technical and administrative support to field staff. There was confusion over roles of the health personnel in the TB programme and the public health facility, as well as poor technical performance. Partnerships were found to be useful in addressing the following areas: (1) the implementation of an external quality assurance scheme for sputum microscopy through involvement of microbiologists from large hospitals and research organizations; (2) training and capacity strengthening of programme and public health facility staff through innovative training and team building exercises organized by the programme, NGOs and the private sector; (3) development of Information, Education and Communication (IEC) material through partnerships with NGOs, and (4) the involvement of local NGOs and private doctors to increase case finding and to improve access to direct observation of treatment (DOT). The paper discusses the lessons learnt in this process and identifies some of the key issues in urban TB control, for consideration by policy makers. PMID- 12758208 TI - Public-private mix DOTS in the Philippines. AB - The National Tuberculosis Prevalence Survey done in 1997 (1997 NPS) revealed that the magnitude of tuberculosis in the Philippines hardly declined between 1982 and 1997. In 1996, the National TB Control Program (NTP) adopted the directly observed treatment short-course (DOTS) strategy. By the end of 2001, more than 90% of the population have access to it. Cohort analysis revealed good treatment outcomes. Despite this improvement in the public sector, there is a concern that the epidemiological impact of DOTS will be limited due to the non-participation of the private practitioners, a major stakeholder. Public-private sector collaboration in TB control was strengthened since the Philippine Coalition Against TB (PhilCAT) was organized in 1994. There are four areas of collaboration, namely, policy development, advocacy and information dissemination, training and research and service delivery. Four models of public private mix (PPM) DOTS in service delivery were developed. The private DOTS clinics provide the space, staff and operational funds while the Department of Health (DOH) provided technical assistance, anti-TB drugs, laboratory supplies and forms. Evaluation showed that PPM in TB control is feasible with good results. The major challenge is to replicate and institutionalize the PPM DOTS. PMID- 12758209 TI - The role of the community in the control of tuberculosis. AB - SETTING: High tuberculosis (TB) prevalence countries, where National TB programmes (NTPs) need to ensure widespread access to effective treatment. OBJECTIVE: To review possible ways in which communities can contribute to TB control and to describe the project "Community TB Care in Africa". DESIGN: A review of the literature on community contribution to TB control and a progress report on the project "Community TB Care in Africa". RESULTS: Eight district based projects in six countries (Botswana, Kenya, Malawi, South Africa, Uganda, Zambia) participated in the overall "Community TB Care in Africa" project. District TB programmes offering patients the choice of community or health facility treatment supervision generally performed effectively, with satisfactory rates of treatment success. Average health system and patient and family costs were lower in comparison with controls. CONCLUSION: Investigators formulated policy recommendations. NTPs should: (1) extend TB care to the community to improve access; (2) identify suitable community TB treatment supporters in consultation with the community; (3) ensure that effective systems extend into the community for recording and reporting, and for supply of anti-TB drugs; (4) monitor community contribution to TB care using standard indicators; (5) develop costed plans for expansion of the community approach. Ministries of Health should: (1) ensure adequate financing; (2) coordinate the efforts of NTPs, donors and NGOs to ensure sustainability; (3) consider opportunities for collaboration between NTPs and HIV/AIDS programmes. PMID- 12758210 TI - Preliminary results of an operational field study to compare side-effects, complaints and treatment results of a single-drug short-course regimen with a four-drug fixed-dose combination (4FDC) regimen in South Sulawesi, Republic of Indonesia. AB - SETTING: Health centres in the South Sulawesi Province, Republic of Indonesia. OBJECTIVES: To compare complaints, side-effects and treatment outcome in new smear-positive patients treated with a single-drug short-course (National TB Programme (NTP)) regimen with those treated with a four-drug fixed-dose combination (4FDC) regimen. DESIGN: A prospective study in which patients are randomly allocated to the NTP or the 4FDC regimen. RESULTS: Preliminary results of the first 360 patients (162 treated with the NTP regimen and 198 with the 4FDC regimen) show that two patients, treated with the NTP regimen, developed jaundice. During the intensive phase of treatment, gastro-intestinal and muscle joint complaints of any duration and gastro-intestinal complaints lasting for 2 consecutive weeks or more were more frequent in patients treated with the NTP regimen. Sputum conversion was 89% in patients treated with the NTP regimen and 94% in those treated with the 4FDC regimen. Nine-five per cent of patients, both regimens, were cured. CONCLUSION: The results so far show that complaints during the intensive phase of treatment are less frequent among patients treated with the 4FDC regimen. The lower dose of pyrazinamide might be the reason. Treatment results are excellent for both regimens. PMID- 12758211 TI - Costs and effectiveness--the impact of economic studies on TB control. AB - This paper assesses the impact of economic studies on TB control during the period 1982-2002, with a focus on cost and cost-effectiveness studies. It begins by identifying broad categories of economic study relevant to TB control, and how economic studies can, theoretically, have an impact on TB control. The impact that economic studies of TB control have had in practice is then analysed through a systematic review of the literature on cost and cost-effectiveness studies related to TB control, and three case studies (one cost study and two cost effectiveness studies). The results show that in the past 20 years, 66 cost effectiveness studies and 31 cost studies have been done on a variety of important TB control topics, with a marked increase occurring after 1994. In terms of numbers, these studies have had most potential for impact in industrialized countries, and within industrialized countries are most likely to have had an impact on policy and practice related to screening and preventive therapy. In developing countries with a high burden of tuberculosis, far fewer studies have been undertaken. Here, the main impact of economic studies has been influencing policy and practice on the use of short-course chemotherapy, justifying the implementation of community-based care in Africa, and helping to mobilize funding for TB control based on the argument that short-course treatment for TB is one of the most cost-effective health interventions available. For the future, cost and cost-effectiveness studies will continue to be relevant, as will other types of economic study. PMID- 12758212 TI - Progress in TB drug development and what is still needed. AB - Highly effective drugs for treating TB were introduced over 30 years ago, yet deaths from the disease continue to increase. New tools are needed, including drugs with activity against multi-drug resistant strains of Mycobacterium tuberculosis. Agents that reduce the duration and complexity of the current therapy would have a major impact on compliance and overall cure rate. In recent years, our understanding of the tubercle bacillus and its interaction with the human host has improved dramatically, particularly with the publication in 1998 of the complete genome sequence of M. tuberculosis H37Rv. New genetic tools have been developed and we can now ascertain the function of individual genes. Thus, many potential drug targets have been identified and a number demonstrated to be essential. Several lead compounds have been found, as well as a potential drug candidate, the nitroimidazopyran PA-824. A far greater effort is needed to translate basic research into drug discovery programmes. High throughput screening and rational design must be employed to find lead compounds acting against well-validated targets and a substantial increase in resources devoted to medicinal chemistry is required to take these leads and turn them into drugs. Models of mycobacterial persistence, in which compounds with potent sterilizing activity can be rapidly analysed, must be characterized. Finally, surrogate markers that give an early indication of treatment outcome would facilitate clinical trials. PMID- 12758213 TI - Pediatric tuberculosis: time for a new approach. PMID- 12758214 TI - Prospects for a better vaccine against tuberculosis. AB - There have been many new promising approaches to developing human vaccines against tuberculosis (TB). Advances in gene and antigen identification, availability of genome sequences, a greater understanding of immune mechanisms in resistance to TB, the development of adjuvants and delivery systems to stimulate T-cell immunity, and increased funding from public and private agencies are some of the reasons for progress in this area. Dozens of vaccine candidates have been tested in animal models in recent years, and several of these are poised to move into clinical trials in the next several years. Thus, there is renewed optimism for the potential of developing new and improved TB vaccines. PMID- 12758215 TI - Comparison of available benchmark dose softwares and models using trichloroethylene as a model substance. AB - By using trichloroethylene as a model substance the U.S. EPA benchmark dose software was compared to the software by Crump and the software by Kalliomaa. Dose-response and dose-effect data on the liver, kidneys, central nervous system (CNS), and tumours were selected for the evaluation. Based on the present study the U.S. EPA software is preferable to the other softwares for dichotomous data. A wider range in benchmark doses was often observed for dichotomous data when the numbers of dose levels were limited. The log-logistic model in most cases gave the best fit when ranking the dichotomous models. In addition, the log-logistic model often implied a more conservative benchmark dose. For continuous data it was more difficult to find a model describing the data. The softwares by Kalliomaa and by the U.S. EPA offered the best opportunities for benchmark dose modelling of continuous data. Flexible models, like the Hill- and the Mult model, are needed for S-shaped continuous data but these models demand more dose levels in order to describe the data. Since the number of dose levels are important for model selection study design is important and should be further evaluated. PMID- 12758216 TI - Some aspects relating to the evaluation of the effects of chemicals on male fertility. AB - Reviews and studies on individual compounds were analyzed as to the suitability of different study designs and endpoints for detecting adverse effects of chemicals on male reproduction in animal species. Of the endpoints investigated, the most sensitive proved to be histopathology of the testes. Using refined histopathology, effects could be detected with a high degree of sensitivity as early as 4 weeks after treatment. Other sensitive endpoints were the weights of reproductive organs, including accessory glands, i.e., testis, epididymis, prostate, and of the seminal vesicle, as well as sperm parameters such as sperm count, sperm morphology, and sperm motility. Sperm motility was found to be in some cases more sensitive than histopathology. The above parameters showed a higher sensitivity than fertility parameters. In fact, in most cases, not only one but several endpoints were affected. Continuous breeding studies and 90-day studies with additional measurements of sperm parameters were similarly effective in detecting compounds which affect male fertility. Interspecies extrapolation factors (IEFs) have been derived for the most sensitive endpoints in laboratory animals. If the calculation is based on caloric demand and a sensitive endpoint of reproductive toxicity, many IEFs tend to be about 1, indicating that humans are generally not more susceptible to reproductive toxicants than laboratory animals. With respect to hazard identification, it is possible to detect adverse effects on male reproduction in a standard subacute study with concentrations that produce significant general toxicity. If effects are found, for the risk assessment the NOAEL has to be determined by testing specific sensitive parameters as specified above. PMID- 12758217 TI - Risk analysis under uncertainty, the precautionary principle, and the new EU chemicals strategy. AB - Three categories of uncertainty in relation to risk assessment are defined; uncertainty in effect, uncertainty in cause, and uncertainty in the relationship between a hypothesised cause and effect. The Precautionary Principle (PP) relates to the third type of uncertainty. Three broad descriptions of the PP are set out, uncertainty justifies action, uncertainty requires action, and uncertainty requires a reversal of the burden of proof for risk assessments. The application of the PP is controversial but what matters in practise is the precautionary action (PA) that follows. The criteria by which the PAs should be judged are detailed. This framework for risk assessment and management under uncertainty is then applied to the envisaged European system for the regulation of chemicals. A new EU regulatory system has been proposed which shifts the burden of proof concerning risk assessments from the regulator to the producer, and embodies the PP in all three of its main regulatory stages. The proposals are critically discussed in relation to three chemicals, namely, atrazine (an endocrine disrupter), cadmium (toxic and possibly carcinogenic), and hydrogen fluoride (a toxic, high-production-volume chemical). Reversing the burden of proof will speed up the regulatory process but the examples demonstrate that applying the PP appropriately, and balancing the countervailing risks and the socio-economic benefits, will continue to be a difficult task for the regulator. The paper concludes with a discussion of the role of precaution in the management of change and of the importance of trust in the effective regulation of uncertain risks. PMID- 12758218 TI - Di-alkyl phosphate biomonitoring data: assessing cumulative exposure to organophosphate pesticides. AB - The 1996 Food Quality Protection Act (FQPA) requires the evaluation of both aggregate and cumulative health risks from pesticides (FFDCA 408(b)(2)(D)(v) and (vi).) Organophosphate (OP) pesticides are the first class of chemicals to undergo FQPA mandated aggregate and cumulative assessments. In this report, summary data on biomonitoring for urinary levels of six alkyl phosphate (AP) metabolites of OPs, as reported in the initial, March 2001, U.S. Centers for Disease Control and Prevention's (CDC) "National Report on Human Exposure to Environmental Chemicals," are compared to EPA modeled estimates of OP exposure reported in Registration Eligibility Decision documents (REDs), Interim REDs and to currently reported cumulative exposure estimates in the EPA's Cumulative Risk Assessment of the Organophosphate Pesticides. This comparison indicates that EPA's aggregate exposure estimates (dietary, drinking water, and non-dietary residential exposures) for many individual OPs were greater than the cumulative estimate for all OPs combined based on the CDC AP biomonitoring data. The results also suggest that EPA's screening level assessments of OPs, while being qualitative indicators of the relative importance of various exposure sources, are not good quantitative indicators of actual exposures. However, the mean biomonitoring estimate of cumulative OP exposure appears to exceed the EPA's subsequent preliminary estimate of cumulative OP exposure by as much as the REDs appear to overestimate the biomonitoring results. While the conservatism, tendency to overestimate exposure, in the individual REDs is readily acknowledged, the conservatism and limitations of applying currently available CDC AP biomonitoring data to evaluate human exposure to OPs are not as readily apparent. We postulate that oral absorption of non-anti cholinergic, pre hydrolyzed OPs, sources of APs other than pesticides, and the conservative result of summing exposure from each AP at the geometric mean contribute to non quantified overestimates of absorbed dosage from the CDC biomonitoring data reported in March 2001. CDC AP biomonitoring data may serve a useful purpose in providing an upper bound estimate of absorbed dosage for "ground truthing" aggregate exposure estimated from first tier models used in REDs, but at best may provide only a credible "target" for the complex cumulative exposure assessment models currently under development. The reliability of quantitative estimates of OP exposure levels will improve as cumulative risk exposure models are validated over time and under use conditions prevalent at the time the AP biomonitoring samples are collected. Analyses contained herein should be revisited and compared to the CDC Second National Report on Human Exposure to Environmental Chemicals ( http://www.cdc.gov/exposurereport), released to the public on January 31, 2003, and the final EPA OP Cumulative Risk Assessment. PMID- 12758219 TI - Ninety-day oral toxicity study of lycopene from Blakeslea trispora in rats. AB - Lycopene, as a suspension in sunflower oil (20% w/w), was tested for subchronic toxicity by administration at dietary concentrations of 0, 0.25, 0.50, and 1.0% to groups of 20 male and 20 female Wistar rats for a period of 90 days. The lycopene examined in this study was derived from a fungal biomass (Blakeslea trispora). Lycopene intake was calculated to be 0, 145, 291, and 586mg/kg body weight/day in control through high-dose males and 0, 156, 312, and 616mg/kg body weight/day in control through high-dose females. The results from this study do not provide any evidence of toxicity of lycopene at dietary levels up to 1.0% as demonstrated by the findings of clinical observations, neurobehavioral observations, motor activity assessment, body weight and food consumption measurements, ophthalmoscopic examinations, hematology, clinical chemistry, urinalysis, organ weights, gross pathology, or histopathology. The No-Observed Effect Level (NOEL) was 1.0% in the diet, the highest dietary concentration tested. PMID- 12758220 TI - Does exposure to bisphenol A represent a human health risk? PMID- 12758222 TI - What's wrong with the National Ambient Air Quality Standard (NAAQS) for fine particulate matter (PM2.5)? PMID- 12758223 TI - Human sex differentiation and its abnormalities. AB - The purpose of this chapter is to review the presentation and management of patients affected by conditions of abnormal sex differentiation. First, the processes of normal sex differentiation are covered, followed by an overview of the various syndromes of abnormal sex differentiation, or intersex conditions, that can occur. These disorders are presented according to the following categories: patients who possess a 46,XX chromosome complement, those who possess a 46,XY chromosome complement, and individuals who present with an atypical sex chromosome complement (i.e. 45,XO or 45,X0/46,XY mosaicism). A description of the medical, surgical and psychological treatment options for people affected by various intersex conditions and reared as females are included. Practice points, based on research studies when available, are dispersed throughout the chapter. Additionally, information pertaining to relevant Internet websites and patient support groups are provided, so that medical staff can educate their patients about the availability of these resources. PMID- 12758224 TI - Congenital malformations of the genital tract and their management. AB - While congenital malformations of the genital tract are not common, the sequelae of their presence can be serious. The practising gynaecologist must be aware of the range of congenital abnormalities that may occur and the symptoms that may result from them. Failure to manage these patients correctly may have long-term sequelae for their psychological, sexual and reproductive health. The involvement of a multi-disciplinary team in dealing with these patients is imperative, and preparation for surgery-particularly in congenital malformations of the vulva and the vagina-is imperative if the long-term sexual function in these patients is to be fulfilled. Surgical correction of vulval abnormalities in adolescence is related solely to sexual function as most of the reconstructive surgery is done in childhood. For the management of Mayer-Rokitansky-Kuster-Hauser (MRKH) syndrome, the recommendation is now that passive dilatation by Franks' technique is the treatment of first choice and only if that fails should surgical approaches be embarked upon. The results of the surgery are similar in all techniques and the particular surgical centre will have its own preference of which technique it adopts. Congenital absence of the cervix is a complex surgical problem and should be dealt with solely in centres with expertise. PMID- 12758225 TI - Disorders of pubertal development. AB - Puberty is the period of life during which reproductive capability is acquired. It is characterized clinically by the acquisition of secondary sexual characteristics associated with a growth spurt, and on average takes 3-4 years. Early maturation is defined as the development of sexual characteristics before the age of 8 years in girls and 9 years in boys. Delayed puberty is defined when there are no signs of puberty at the age of 13.4 years in girls and 14 years in boys (2 SD above the mean of chronological age for the onset of puberty). There are many forms of premature sexual maturation: gonadotrophin-dependent (central, or 'idiopathic' or 'true' precocious puberty) and gonadotrophin-independent precocious puberty (McCune-Albright syndrome in girls, testotoxicosis in boys); isolated premature thelarche (in the forms of classical, atypical and variant); premature adrenarche (characterized by the production of significant quantities of androgens between 5 and 8 years of age); premature menarche. The differential diagnosis of delayed puberty is between constitutional delay of growth and puberty, pubertal delay secondary to chronic disease and hypogonadotrophic hypogonadism. PMID- 12758226 TI - Eating disorders in adolescence and their sequelae. AB - Eating disorders are prevalent in adolescents and are associated with significant medical and psychiatric morbidity. Amenorrhoea, one of the cardinal features of anorexia nervosa, is the most likely reason for consulting the gynaecologist. Amenorrhoea in a young woman should alert the gynaecologist to the possibility of an underlying eating disorder. Osteopenia is a potentially irreversible complication of prolonged amenorrhoea and a low oestrogen state. Eating disorders are best managed by a team approach, with the team comprising a physician, nutritionist and therapist. Oestrogen replacement therapy has not been shown to be an effective treatment for osteopenia in anorexia nervosa and the gynaecologist should avoid simply prescribing oestrogen replacement therapy without referring the patient for comprehensive treatment of the eating disorder. Nutritional rehabilitation, weight restoration and resumption of spontaneous menses are the mainstay of medical management. Calcium and vitamin D supplementation and moderate weight-bearing exercise should be prescribed where indicated. Newer therapeutic options for the treatment of osteopenia include DHEA, IGF-1 and alendronate. PMID- 12758227 TI - Menstrual disorders in adolescence. AB - Problems associated with menstruation affect 75% of adolescent females and are a leading reason for visits to physicians. This chapter begins with a review of the timing and characteristics of normal menstruation during adolescence. It then discusses the evaluation and management of adolescents with amenorrhoea, dysmenorrhoea and abnormal uterine bleeding. An approach to adolescent amenorrhoea is presented that utilizes primary versus secondary amenorrhoea, delayed versus normal pubertal development, and the presence or absence of hyperandrogenism as nodal points for decision making. The differential diagnosis of dysmenorrhoea and the management of primary dysmenorrhoea and endometriosis are reviewed. The section on abnormal uterine bleeding contrasts anovulatory dysfunctional uterine bleeding (DUB) with bleeding secondary to problems of pregnancy, uterine pathology, exogenous hormone use and systemic bleeding disorders. PMID- 12758228 TI - Adolescent pelvic pain. AB - Chronic pelvic pain is a frequent complaint in adolescent females. It is a complex disorder with multiple causes. The assessment must attempt to differentiate between gynaecological and non-gynaecological sources of pain. An understanding of the physical, cognitive and environmental factors associated with the pain are essential. Laparoscopy has been used in the assessment of CPP but a significant number of patients will have no obvious aetiology at the time of laparoscopy. For the young patient with CPP, a multidisciplinary approach may be essential to facilitate diagnosis and management. Although the symptoms may not always be curable, management that allows the young female to obtain normal or near normal function may be possible. This chapter focuses on the various causes of pelvic pain in the adolescent female with a focus on the assessment, diagnosis and treatment of the different causes. PMID- 12758229 TI - Sexual function, sexual abuse and sexually transmitted diseases in adolescence. AB - As adolescents progress through puberty, many biological changes occur and, for young women, this includes the onset of menses and the capability for reproduction. During this time, sexual identity is developed and expressions of sexuality become more frequent. Adolescent women engage in a variety of sexual behaviours, both non-coital and coital. As teens begin dating relationships, they are at risk for dating violence and sexual abuse. Some may even be raped after sedation with a 'date rape' drug. As adolescents attempt to develop intimate sexual relationships, they may be at high risk for health consequences associated with sexual activity, such as pregnancy and sexually transmitted diseases (STDs). Providers, such as physicians, nurse practitioners, physician assistants and nurses, must know current STD diagnosis and treatment recommendations to decrease morbidity caused by these infections. By knowing how to interview, understanding legal issues and anticipating concerns pertinent to teens, providers have the opportunity to decrease barriers to health care for adolescents. PMID- 12758230 TI - Unplanned teenage pregnancy. AB - This chapter addresses unplanned teenage pregnancy from a human-rights-based perspective. Many programmes focus only on the negative aspects of young people's sexual and reproductive health; putting rights at the centre of teenage sexual health avoids treating adolescents as a homogeneous collection of discrete problems. Taking a rights-based approach to adolescent sexual and reproductive health encompasses the inter-relationships and complexity of factors influencing choices and decisions. In this chapter we explore the meaning of a rights-based approach and examine its implementation in the provision of sexuality, education and health services. We provide examples of international strategies for ensuring the participation of diverse young people and generating youth-friendly services to meet their sexual and reproductive needs and rights. PMID- 12758231 TI - Acne and hirsuties in teenagers. AB - Acne and body hair are both cutaneous responses to androgenic stimulation. They are normal events in adolescent girls. There is considerable variation in the evolution of the two conditions. The sebaceous gland is exquisitely sensitive to androgens, and acne appears with the onset of puberty, peaks in prevalence in the teenage years and gradually improves thereafter. Hair growth on the face, trunk and limbs develops more slowly and generally peaks in the 20s. Indications for endocrine investigation include very severe acne, onset of acne and hirsuties in the very early stage of puberty (Tanner stage 3) and systemic virilism. Treatment for acne and hirsuties can be either topical or systemic. The choice of therapy is based on the severity of the disease rather than the results of endocrine investigation. Further, since PCO is related to impaired glucose tolerance, advice relating to lifestyle changes should be offered to prevent the development of diabetes. PMID- 12758232 TI - Pelvic tumours in adolescence. AB - Malignant pelvic tumours in adolescents are rare. Cancers most commonly associated with adolescent women are ovarian germ-cell tumours, ovarian stromal tumours, genital rhabdomyosarcomas and cervico-vaginal clear-cell adenocarcinomas. The incidence of the last of these has reduced with the abandonment of diethylstilbestrol (stilboestrol) therapy in pregnancy. With sexual activity among adolescent women increasing, the incidence of cervical cancer and gestational trophoblastic tumours is rising. Treatment for pelvic cancers in adolescence should be in a multidisciplinary setting and in most cases surgery should be conservative with the aim of preserving sexual and reproductive function. With a few notable exceptions, the prognosis for most malignant pelvic tumours that occur in adolescence is good and treatment is with curative intent. PMID- 12758234 TI - Cancers of the head and neck region in developing countries. PMID- 12758235 TI - Tumor hypoxia at the micro-regional level: clinical relevance and predictive value of exogenous and endogenous hypoxic cell markers. AB - BACKGROUND AND PURPOSE: Tumor oxygenation is recognized as an important determinant of the outcome of radiotherapy and possibly also of other treatment modalities in a number of tumor types and in particular in squamous cell carcinomas. The hypoxic status of various solid tumors has been related to a poor prognosis due to tumor progression towards a more malignant phenotype, with increased metastatic potential, and an increased resistance to treatment. It has been demonstrated in head and neck cancer that hypoxic radioresistance can be successfully counteracted by hypoxia modifying approaches. The microregional distribution and the level of tumor hypoxia depend on oxygen consumption and temporal and spatial variations in blood supply. It is unclear if severely hypoxic cells can resume clonogenicity when O(2) and nutrients become available again as a result of (treatment related) changes in the tumor microenvironment. Non-terminally differentiated hypoxic cells that are capable of proliferation are important for outcome because of their resistance to radiotherapy and possibly other cytotoxic treatments. Various exogenous and endogenous markers for hypoxia are currently available and can be studied in relation to each other, the tumor architecture and the tumor microenvironment. Use of nitroimidazole markers with immunohistochemical detection allows studying tumor cell hypoxia at the microscopic level. Co-registration with other microenvironmental parameters, such as vascular architecture (vascular density), blood perfusion, tumor cell proliferation and apoptosis, offers the possibility to obtain a comprehensive functional image of tumor patho-physiology and to study the effects of different modalities of cancer treatment. CONCLUSION: A number of functional microregional parameters have emerged that are good candidates for future use as indicators of tumor aggressiveness and treatment response. The key question is whether these parameters can be used as tools for selection of treatment strategies for individual patients. This requires testing of these markers in prospective randomized clinical trials comparing standard treatment against experimental treatments targeting the relevant microregional constituent. PMID- 12758237 TI - Quantitative tissue perfusion measurements in head and neck carcinoma patients before and during radiation therapy with a non-invasive MR imaging spin-labeling technique. AB - PURPOSE: Tumor blood flow, tumor tissue perfusion and oxygen supply have substantial influence on the responsiveness of tumors to radiotherapy. This study was aimed at implementing and evaluating a non-invasive functional magnetic resonance (MR) imaging spin-labeling technique at a main magnetic field strength of 2T for measuring tissue perfusion changes in head and neck carcinoma patients before and during radiotherapy. METHODS: Tissue perfusion was determined quantitatively in ten patients with head and neck cancer. Five patients were investigated twice during radiation therapy. For perfusion measurements, a non invasive MR spin-labeling technique was employed: The longitudinal relaxation time T(1) was measured with segmented Snapshot-FLASH imaging after either slice selective or non-selective spin inversion. Perfusion values were calculated pixelwise employing a two-compartment tissue model. With this technique no contrast agents are required so that repetitive measurements are possible. Perfusion images with a slice thickness of 10mm and an in-plane resolution of 1.9x2.8mm(2) were acquired at a total scan time of 8:30min per scan. RESULTS: With the non-invasive MR imaging technique it was possible to visualize tumor and normal tissue perfusion as well as perfusion changes in the course of radiotherapy with a spatial resolution of less than 3mm. Among the investigated subjects measured tumor perfusion and changes in perfusion were heterogenous. In 4/5 patients studied at the start and end of radiotherapy, perfusion decreased, while in one patient there was an increase. CONCLUSIONS: A method is presented that allows non-invasive and repetitive characterization of tissue perfusion. This parameter may be used for treatment stratification, especially in treatments that use vasomodulation or anti-angiogenic agents. PMID- 12758236 TI - Radiotherapy with or without mitomycin c in the treatment of locally advanced head and neck cancer: results of the IAEA multicentre randomised trial. AB - BACKGROUND AND PURPOSE: Single agent mitomycin c (MMC) has been shown to improve the outcome of radiotherapy in single institution trials. In order to confirm these findings in a broader worldwide setting, the International Atomic Energy Agency (IAEA) initiated a multicentre trial randomising between radiotherapy alone versus radiotherapy plus MMC. MATERIAL AND METHODS: Patients with advanced head and neck cancer were treated with primary curative radiotherapy (66 Gy in 33 fractions with five fractions per week) +/-a single injection (15 mg/m(2)) of MMC at the end of the first week of radiotherapy. Stratification parameters were tumour localization, T-stage, N-stage, and institution. A total of 558 patients were recruited in the trial from February 1996 to December 1999. Insufficient accrual and reporting led to the exclusion of three centres. The final study population consisted of 478 patients from seven centres. Patients had stage III (n=223) or stage IV (n=255) squamous cell carcinoma of the oral cavity (n=230), oropharynx (n=140), hypopharynx (n=65) or larynx (n=43). Prognostic factors like age, gender, site, size, differentiation and stage were well balanced between the two arms. RESULTS: The haematological side effects of MMC were very modest (<5% grade 3-4) and did not require any specific interventions. Furthermore, MMC did not enhance the incidence or severity of acute and late radiation side effects. Confluent mucositis and dry skin desquamation was common, occurring in 56% and 62% of patients, respectively. The overall 3-year primary locoregional tumour control, disease-specific and overall survival rates were 19, 36 and 30%, respectively. Gender, haemoglobin drop, tumour site, tumour and nodal stage were significant parameters for loco-regional tumour control. There was no significant effect of MMC on locoregional control or survival, except for the 161 N0 patients, where MMC resulted in a better loco-regional control (3-year estimate 16% vs. 29%, P=0.01). CONCLUSIONS: The study did not show any major influence of MMC on loco-regional tumour control, survival or morbidity after primary radiotherapy in stage III-IV head and neck cancer except in N0 patients where loco-regional control was significantly improved. PMID- 12758238 TI - Measurements of hypoxia using pimonidazole and polarographic oxygen-sensitive electrodes in human cervix carcinomas. AB - BACKGROUND AND PURPOSE: The measurement of tumour oxygenation using Eppendorf oxygen-sensitive needle electrodes can provide prognostic information but the method is limited to accessible tumours that are suitable for electrode insertion. In this paper the aim was to study the relationship between such physiological measurements of tumour hypoxia and the labelling of tumours with the hypoxia-specific marker pimonidazole. MATERIALS AND METHODS: Assessment of tumour oxygen partial pressure (pO(2)) using an Eppendorf pO(2) histograph and immunohistochemical pimonidazole labelling was carried out in 86 patients with primary cervix carcinomas. Pimonidazole was given as a single injection (0.5 g/m(2) i.v.) and 10-24 h later pO(2) measurements were made and biopsies taken. Tumour oxygenation status was evaluated as the median tumour pO(2) and the fraction of pO(2) values 0.05). CONCLUSION: HDR brachytherapy in the management of the cervix appears to be a safe and efficacious approach. Pelvic control, survival and complications rates are quite similar when compared with LDR. PMID- 12758246 TI - To what extent can digital images obtained with a non-isocentric C-arm be used for brachytherapy treatment planning in gynaecology. AB - BACKGROUND AND PURPOSE: The non-isocentric C-arm X-ray fluoroscopy unit is not typically used as a brachytherapy localizer. The main aim of the study is to examine the extent to which the potential of a mobile C-arm unit as a brachytherapy localizer for gynaecological treatments with Fletcher-Suit applicator can be used. PATIENTS AND METHODS: With a C-arm, located in the brachytherapy treatment room in situ reconstruction images are made, eliminating the transportation of the patient to the X-ray room. The brachytherapy treatment planning is based on the frame grabbed images using variable angle reconstruction. RESULTS: The method of making a pair of reconstruction images with the C-arm is described. The accuracy of the reconstruction has been examined with phantom studies and therapy plans of Fletcher-Suit applications of different variable angles. PMID- 12758247 TI - Clinical evaluation of dynamic arc conformal radiotherapy for paraaortic lymph node metastasis. AB - PURPOSE: This study was performed to evaluate the efficacy and safety of dynamic arc conformal radiotherapy, a simple intensity modulated radiation therapy (IMRT), for the treatment of paraaortic lymph node metastases. MATERIALS AND METHODS: Twenty-nine patients with paraaortic lymph node metastases were enrolled in this study. The total planned dose was 55-60 Gy. A computed tomography (CT) simulator was used in the treatment planning. RESULTS: The total radiation dose delivered was 50-63.4 Gy (median 60 Gy). Sixteen of 29 patients showed local tumor shrinkage on CT, and the 2 year in-field recurrence free survival rate was 58%. Acute Grade 1 and Grade 2 gastrointestinal disorders occurred in 31% and 17%, respectively, and acute Grade 2 liver dysfunction occurred in 7%. As a late complication, Grade 1 and Grade 2 liver dysfunction occurred in six patients (21%) and five patients (17%), respectively. There was no renal dysfunction or myelopathy detected. CONCLUSION: Dynamic arc conformal radiotherapy, a simple IMRT, is a safe and effective treatment method for paraaortic lymph node metastasis. PMID- 12758248 TI - Application of monomer/polymer gel dosimetry to study the effects of tissue inhomogeneities on intensity-modulated radiation therapy (IMRT) dose distributions. AB - BACKGROUND AND PURPOSE: When planning an intensity-modulated radiation therapy (IMRT) treatment in a heterogeneous region (e.g. the thorax), the dose computation algorithm of a treatment planning system may need to account for these inhomogeneities in order to obtain a reliable prediction of the dose distribution. An accurate dose verification technique such as monomer/polymer gel dosimetry is suggested to verify the outcome of the planning system. MATERIALS AND METHODS: The effects of low-density structures: (a) on narrow high-energy (18 MV) photon beams; and (b) on a class-solution IMRT treatment delivered to a thorax phantom have been examined using gel dosimetry. The used phantom contained air cavities that could be filled with water to simulate a homogeneous or heterogeneous configuration. The IMRT treatment for centrally located lung tumors was delivered on both cases, and gel derived dose maps were compared with computations by both the GRATIS and Helax-TMS planning system. RESULTS: Dose rebuildup due to electronic disequilibrium in a narrow photon beam is demonstrated. The gel measurements showed good agreement with diamond detector measurements. Agreement between measured IMRT dose maps and dose computations was demonstrated by several quantitative techniques. An underdosage of the planning target volume (PTV) was revealed. The homogeneity of the phantom had only a minor influence on the dose distribution in the PTV. An expansion of low-level isodoses in the lung volume was predicted by collapsed cone computations in the heterogeneous case. CONCLUSIONS: For the class-solution described, the dose in centrally located mediastinal tumors can be computed with sufficient accuracy, even when neglecting the lower lung density. Polymer gel dosimetry proved to be a valuable technique to verify dose calculation algorithms for IMRT in 3D in heterogeneous configurations. PMID- 12758249 TI - Quality assurance of a system for improved target localization and patient set-up that combines real-time infrared tracking and stereoscopic X-ray imaging. AB - BACKGROUND AND PURPOSE: The aim of this study is to investigate the positional accuracy of a prototype X-ray imaging tool in combination with a real-time infrared tracking device allowing automated patient set-up in three dimensions. MATERIAL AND METHODS: A prototype X-ray imaging tool has been integrated with a commercially released real-time infrared tracking device. The system, consisting of two X-ray tubes mounted to the ceiling and a centrally located amorphous silicon detector has been developed for automated patient positioning from outside the treatment room prior to treatment. Two major functions are supported: (a) automated fusion of the actual treatment images with digitally reconstructed radiographs (DRRs) representing the desired position; (b) matching of implanted radio opaque markers. Measurements of known translational (up to 30.0mm) and rotational (up to 4.0 degrees ) set-up errors in three dimensions as well as hidden target tests have been performed on anthropomorphic phantoms. RESULTS: The system's accuracy can be represented with the mean three-dimensional displacement vector, which yielded 0.6mm (with an overall SD of 0.9mm) for the fusion of DRRs and X-ray images. Average deviations between known translational errors and calculations varied from -0.3 to 0.6mm with a standard deviation in the range of 0.6-1.2mm. The marker matching algorithm yielded a three-dimensional uncertainty of 0.3mm (overall SD: 0.4mm), with averages ranging from 0.0 to 0.3mm and a standard deviation in the range between 0.3 and 0.4mm. CONCLUSIONS: The stereoscopic X-ray imaging device integrated with the real-time infrared tracking device represents a positioning tool allowing for the geometrical accuracy that is required for conformal radiation therapy of abdominal and pelvic lesions, within an acceptable time-frame. PMID- 12758250 TI - Development of a F actin-based live-cell fluorimetric microplate assay for diarrhetic shellfish toxins. AB - A new cytotoxicity assay for detection and quantitation of diarrhetic shellfish toxins (DSP) is presented. This assay is based upon fluorimetric determination of F-actin depolymerization induced by okadaic acid (OA)-class compounds in the BE(2)-M17 neuroblastoma cell line. No interferences were observed with other marine toxins such as saxitoxin, domoic acid, or yessotoxin, thus indicating a good specificity of the assay as expected by the direct relationship between protein phosphatase inhibition and cytoskeletal changes. The proposed method is rapid (<2h) and shows a linear response in the range of 50-300 nM OA. The detection limit of the assay for crude methanolic extracts of bivalves lies between 0.2 and 1.0 microg OA per gram of digestive glands, depending on the type of samples (fresh or canned), thus being similar to that of the mouse bioassay. The performance of this assay has been evaluated by comparative analysis of 32 toxic mussel samples by the F-actin assay, mouse bioassay, HPLC and PP2A inhibition assay. Results obtained by the F-actin method showed no differences with HPLC and significant correlation with PP2A inhibition assay (r(2)=0.71). No false negative results were obtained with this new cell assay, which also showed optimum reproducibility. PMID- 12758251 TI - Fluorescence spectral properties of cyanine dye-labeled DNA oligomers on surfaces coated with silver particles. AB - We examined the fluorescence spectral properties of DNA oligomers, labeled with Cy3 or Cy5, when bound to quartz surfaces coated with metallic silver particles. Prior to binding of labeled DNA the surfaces were treated with polylysine or 3 aminopropyl triethoxysilane or were coated with avidin for binding of biotinylated oligomers. The fluorescence intensities were increased an average of 8-fold on these surfaces. Despite the increased emission intensity, the photostability of the labeled DNA was the same or higher on the silver-coated surfaces than on the uncoated slides. The time-integrated intensities, that is the area under the intensity plots with continuous illumination, increased an average of 6-fold. In all cases the lifetimes were dramatically shortened on the silver particles, indicating an over 100-fold increase in the radiative decay rates. These results suggest the use of substrates containing silver particles for increased sensitivity of DNA detection on DNA arrays. PMID- 12758252 TI - Detection of JNK and p38 activation by flow cytometry analysis. AB - JNK and p38 protein kinases are involved in the signal transduction of apoptotic stimulus. JNK and p38 are activated by dual phosphorylation on threonine and tyrosine residues. Different techniques such as Western blotting (WB) and confocal microscopy analysis have been developed to detect the activation by using antibodies that recognize the phosphorylated forms of both enzymes. However, these techniques are time consuming, not quantitative, and dependent on subjective interpretation. Herein, we describe a flow cytometry-based analysis to detect JNK and p38 activation. Using human primary lymphocytes and Jurkat CD4(+) T cells stimulated with PMA/ionomycin, we demonstrate activation (phosphorylation) of JNK and p38, which is further confirmed by two additional established techniques (WB and confocal microscopy). Flow cytometry analysis is shown to be more sensitive than WB to detect JNK and p38 activation, which can be quantitated and enables us to study their activation within cell populations. PMID- 12758254 TI - Single-nucleotide polymorphism detection in plants using a single-stranded pyrosequencing protocol with a universal biotinylated primer. AB - Analysis of variations in plant genomes is increasingly focused on single nucleotide polymorphism (SNP) analysis, increasing the need for fast yet reliable, simple, and cost-effective techniques to handle the large number of these polymorphisms within large plant genomes. Pyrosequencing technology offers a technique that takes advantage of the interaction of four enzymes in a single tube assay to measure DNA synthesis in real time. Pyrosequencing provides a DNA sequence and an advantage over alternative techniques in poorly characterized genomes such as those of most plant species. Here we compare the use of both single-stranded and double-stranded template Pyrosequencing on plant tissue for SNP identification. Different enzymatic strategies for double-stranded template preparation were compared. Preparation of double-stranded template from plant tissue required labor-intensive purification to allow double-stranded Pyrosequencing. A more cost-effective and less labor-intensive alternative to double-stranded template preparation in plants has been developed using a universal biotinylated primer to improve the efficiency of single-stranded Pyrosequencing. This provides an efficient high-throughput method for SNP analysis by Pyrosequencing. PMID- 12758253 TI - A high-throughput solid-phase extraction assay capable of measuring diverse polyprenyl phosphate: sugar-1-phosphate transferases as exemplified by the WecA, MraY, and MurG proteins. AB - The bacterial proteins WecA and MraY are members of the polyprenyl phosphate:N acetylhexosamine-1-phosphate transferase family, each of which catalyzes the transfer of a specific hexosamine 1-P from a soluble UDP-hexosamine substrate to a bactoprenyl phosphate carrier at the membrane surface. Currently, assays designed to quantitate the activity of these enzymes rely on paper chromatography or liquid-liquid extractions or are specialized to a few members of the family. We describe a generalizable, high-throughput, one-pot assay for these activities that uses a solid-liquid bead-based separation system to selectively adsorb the highly hydrophobic products of reaction. By judicious choice of radiolabeled UDP hexosamine precursor, the same format can be used to quantitate not only diverse members of this transferase family, but also enzymes that catalyze the further modification of these transferase products. This possibility is exemplified by the MurG protein of bacterial cell wall synthesis, which catalyzes the addition of an N-acetylglucosamine residue to the product of the MraY reaction. Thus, the use of this flexible assay tool will allow a critical biochemical and enzymologic analysis of many such membrane-bound transferases in a similar setting. PMID- 12758255 TI - Quantitative analysis of membrane protein-amphiphile interactions using resonance energy transfer. AB - This work describes a simple method for determining the association constant of amphiphiles to membrane proteins. The method uses a fluorescent phospholipid probe, which senses the competition among unlabeled amphiphiles for positions on the transmembrane surface of the protein. The contact between the probe and the protein surface is detected through resonance energy transfer. We have analyzed theoretically this process deriving a general equation for the dependence of the energy transfer efficiency on the composition of the micelles/bilayers in which the protein is inserted. This equation includes an exchange constant for each amphiphile, which gives a measure of its affinity for the protein with respect to that of an amphiphile set as the reference. We applied this method to determine the exchange constant of different phospholipids for the plasma membrane calcium pump. PMID- 12758256 TI - Detection of farnesyl diphosphate accumulation in yeast ERG9 mutants. AB - A sensitive method was developed for measuring farnesyl diphosphate (FPP) accumulation in a mutant strain of Saccharomyces cerevisiae. The strain was blocked at squalene synthase (ERG9 gene) in the isoprenoid pathway and had the catalytic domain of the 3-hydroxy-3-methylglutaryl coenzyme A reductase gene integrated into the chromosome. It required ergosterol for growth and produced E,E-farnesol. The method was based on the isolation of FPP using the anion exchanger Macro Prep High Q and conversion of FPP to E,E-farnesol with alkaline phosphatase. Farnesol was measured using gas chromatography-mass spectrometry. Background farnesol in the cell-free extract was also retained by the anion exchanger, but was removed with repeated washing with methanol. Both 1M NaCl and 40% (v/v) methanol were required in the elution buffer to effectively elute FPP. The preparation of cell-free extract in Bis-Tris propane/HCl, pH 7, buffer containing 0.025% (w/v) Triton X-100 and 15 mM MgCl(2) provided optimum conditions for the stabilization of FPP. PMID- 12758257 TI - Modeling of DNA hybridization kinetics for spatially resolved biochips. AB - The marriage of microfluidics with detection technologies that rely on highly selective nucleic acid hybridization will provide improvements in bioanalytical methods for purposes such as detection of pathogens or mutations and drug screening. The capability to deliver samples in a controlled manner across a two dimensional hybridization detection platform represents a substantial technical challenge in the development of quantitative and reusable biochips. General theoretical and numerical models of heterogeneous hybridization kinetics are required in order to design and optimize such biochips and to develop a quantitative method for online interpretation of experimental results. In this work we propose a general kinetic model of heterogeneous hybridization and develop a technique for estimating the kinetic coefficients for the case of well spaced, noninteracting surface-bound probes. The experimentally verified model is then incorporated into the BLOCS (biolab-on-a-chip simulation) 3D microfluidics finite element code and used to model the dynamic hybridization on a biochip surface in the presence of a temperature gradient. These simulations demonstrate how such a device can be used to discriminate between fully complementary and single-base-pair mismatched hybridization using fluorescence detection by interpretation of the unique spatially resolved intensity pattern. It is also shown how the dynamic transport of the targets is likely to affect the rate and location of hybridization as well as that, although nonspecific hybridization is present, the change in the concentration of hybridized targets over the sensor platform is sufficiently high to determine if a fully complementary match is present. Practical design information such as the optimum transport speed, target concentration, and channel height is presented. The results presented here will aid in the interpretation of results obtained with such a temperature-gradient biochip. PMID- 12758258 TI - Identification and quantification of mutagenic halogenated cytosines by gas chromatography, fast atom bombardment, and electrospray ionization tandem mass spectrometry. AB - Oxidative modification of nucleic acids has been implicated in carcinogenesis. One potential mechanism involves halogenation by the myeloperoxidase and eosinophil peroxidase systems of phagocytes. In the current studies, three mass spectrometric methods for the in vitro and in vivo analysis of halogenated cytosines and deoxycytidines were compared: gas chromatography-electron ionization-mass spectrometry (GC-EI-MS) with a quadrupole instrument, fast atom bombardment or electrospray ionization (ESI) tandem MS with a four-sector magnetic instrument, and liquid chromatography ESI tandem MS (HPLC-ESI-MS/MS) with an ion-trap instrument. GC-EI-MS with selected ion monitoring of dimethyl tert-butylsilyl derivatives of nucleobases was the most sensitive method. High energy collisionally induced dissociation MS/MS analysis with a four-sector magnetic instrument yielded detailed structural information about halogenated nucleoside adducts but required relatively large amounts of material. The most sensitive analysis of intact halogenated deoxycytidine was achieved with extracted ion chromatograms using HPLC-ESI-MS/MS with an ion-trap instrument. Our results indicate that GC-EI-MS is the methodology of choice for ultrasensitive analysis of halogenated cytosines. HPLC-ESI-MS/MS provides greater structural detail for these compounds and may rival GC-EI-MS in sensitivity with more advanced liquid chromatography applications. The mass spectrometric methods we have developed should be useful for evaluating the role of phagocyte-derived oxidants in halogenating nucleobases, nucleosides, and DNA at sites of inflammation. PMID- 12758259 TI - A homogeneous, nonradioactive high-throughput fluorogenic protein kinase assay. AB - Protein kinases play an important role in many cellular processes and mediate cellular responses to a variety of extracellular stimuli. They have been identified by many pharmaceuticals as valid targets for drug discovery. Because of the large number of protein kinases, and the large number of compounds to be screened, it is important to develop assay systems that are not only sensitive but also homogeneous, fast, simple, nonradioactive, and cost-effective. Here we present a novel, rapid, robust assay to measure the enzyme activity of low concentrations of several serine/threonine and tyrosine protein kinases. It is based on the use of fluorogenic peptide substrates (Rhodamine 110, bis peptide amide) that are cleaved before phosphorylation to release the free Rhodamine 110; upon phosphorylation, cleavage is hindered, and the compound remains as a nonfluorescent peptide conjugate. The assay can be carried out in single- as well as multiwell plate formats such as 96- and 384-well plates. The signal-to-noise ratio is very high (40), the Z(') is over 0.8, and the signal is stable for at least 4h. Finally, the assay is easily adapted to a robotic system for drug discovery programs targeting protein kinases. PMID- 12758260 TI - Validation of an algorithm for automatic quantification of nucleic acid copy numbers by real-time polymerase chain reaction. AB - Real-time quantitative polymerase chain reaction (PCR) with on-line fluorescence detection has become an important technique not only for determination of the absolute or relative copy number of nucleic acids but also for mutation detection, which is usually done by measuring melting curves. Optimum assay conditions have been established for a variety of targets and experimental setups, but only limited attention has been directed to data evaluation and validation of the results. In this work, algorithms for the processing of real time PCR data are evaluated for several target sequences (p53, IGF-1, PAI-1, Factor VIIc) and compared to the results obtained by standard procedures. The algorithms are implemented in software called SoFAR, which allows fully automatic analysis of real-time PCR data obtained with a Roche LightCycler instrument. The software yields results with considerably increased precision and accuracy of quantifications. This is achieved mainly by the correction of amplification independent signal trends and a robust fit of the exponential phase of the signal curves. The melting curve data are corrected for signal changes not due to the melting process and are smoothed by fitting cubic splines. Therefore, sensitivity, resolution, and accuracy of melting curve analyses are improved. PMID- 12758261 TI - Merging fluorescence resonance energy transfer and expressed protein ligation to analyze protein-protein interactions. AB - Determination of protein oligomerization state can be technically challenging. We have combined the methods of expressed protein ligation (EPL) and fluorescence resonance energy transfer (FRET) for the analysis of protein homo-oligomerization states. We have attached fluorescein (donor) and rhodamine (acceptor) chromophores via dipeptide linkages to the C-termini of three recombinant proteins and examined the potential for FRET between mixtures of these semisynthetic proteins. The known protein dimer (glutathione S-transferase) showed evidence of FRET and the known protein monomer (SH2 domain phosphatase-1) did not display FRET. Using this method, the previously uncharacterized circadian rhythm enzyme, serotonin N-acetyltransferase, displayed significant FRET, indicating its likely propensity for dimerization or more complex oligomerization. These results establish the potential of the union of EPL and FRET in the analysis of protein-protein interactions and provide insight into the unusual enzymatic behavior of a key circadian rhythm enzyme. PMID- 12758262 TI - Enhancement of the fluorescence and stability of o-phthalaldehyde-derived isoindoles of amino acids using hydroxypropyl-beta-cyclodextrin. AB - Addition of hydroxypropyl-beta-cyclodextrin to o-phthalaldehyde (OPA)-amino acid thiol reaction mixtures is shown to cause significant enhancement of the fluorescence of the isoindole product for a wide range of amino acids, with the largest effects observed in the cases of glycine and lysine. The largest enhancement observed was a factor of 2.67 in the case of the derivative of glycine. This fluorescence enhancement is the result of the formation of a 1:1 host:guest inclusion complex between the isoindole and the cyclodextrin. Relatively small association constants of 44 and 130 M(-1) were obtained for the inclusion of the derivatives of glycine and lysine, respectively. Inclusion of the isoindole derivative into hydroxypropyl-beta-cyclodextrin was also found to result in a significant stabilization of the isoindole derivatives, contrary to what has been previously reported for inclusion into beta-cyclodextrin. For example, the lifetime of the lysine derivative was found to increase from 42 to 222 min, a factor of 5.3. These results have potential applications in fluorescence-based HPLC and high-performance capillary electrophoresis amino acid analysis methods using OPA derivation. Addition of hydroxypropyl-beta cyclodextrin to the reaction mixture results in an increase in both the fluorescence and the stability of the isoindole product, providing potentially significant improvements to the method. PMID- 12758263 TI - A new approach to monitoring proteolysis phenomena using antibodies specifically directed against the enzyme cleavage site on its substrate. AB - Proteolysis is a generic biochemical process that is central in all biological activities. A new strategy for monitoring this biochemical process is proposed here. This approach is based on the production of rabbit polyclonal anti-peptide antibodies directly against the cleavage site on the substrate of the enzyme responsible for proteolysis. So long as the molecule's cleavage site is intact, the antibody will bind to the protein. However, after cleavage of the peptide bond by the protease, the antibody will no longer be able to recognize the substrate. Thus, the development of an ELISA that uses this specific antibody allows hydrolysis of the substrate protein to be monitored. Hydrolysis of beta casein by plasmin, the main indigenous protease of milk, during the ripening of Swiss-type cheese, has been chosen as a model for this study. PMID- 12758264 TI - Sequential one-step extraction and analysis of triacylglycerols and fatty acids in plant tissues. AB - A method for plant tissue digestion and triacylglycerol (TAG) extraction followed by transmethylation of TAGs to produce the fatty acid methyl esters (FAMEs) from small storage tissue samples is presented. The method allows the analysis of both TAGs and FAMEs from the same sample. Several reagent mixtures and different experimental conditions were tested on sliced sunflower seeds. The best results were obtained using a mixture that was 33.3% a solution of NaCl (0.17 M) in methanol and 66.6% heptane by volume. The TAGs in the heptane solution were transmethylated with a mixture containing methanol:toluene:dimethoxypropane:H(4)SO(2) (39:20:5:2, by vol). The method was also tested on other oil seed storage tissue (soybean) and fruit tissues from olive and acorn. In all cases, sunflower, soybean, olive, and acorn, the TAGs and FAMEs composition data obtained by this method were quite similar to data from a standard analysis method. In samples with high protein content, such as soybean and sunflower seeds, the TAG extraction was incomplete. The water content of fruit samples did not interfere with TAG extraction obtained by this method. PMID- 12758265 TI - Expression screen by enzyme-linked immunofiltration assay designed for high throughput purification of affinity-tagged proteins. AB - High-throughput purification of affinity-tagged fusion proteins is currently one of the fastest developing areas of molecular proteomics. A prerequisite for success in protein purification is sufficient soluble protein expression of the target protein in a heterologous host. Hence, a fast and quantitative evaluation of the soluble-protein levels in an expression system is one of the key steps in the entire process. Here we describe a high-throughput expression screen for affinity-tagged fusion proteins based on an enzyme linked immunofiltration assay (ELIFA). An aliquot of a crude Escherichia coli extract containing the analyte, an affinity-tagged protein, is adsorbed onto the membrane. Subsequent binding of specific antibodies followed by binding of a secondary antibody horseradish peroxidase (HRP) complex then allows quantitative evaluation of the analyte using tetramethylbenzidine as the substrate for HRP. The method is accurate and quantitative, as shown by comparison with results from western blotting and an enzymatic glutathione S-transferase (GST) assay. Furthermore, it is a far more rapid assay and less cumbersome than western blotting, lending itself more readily to high-throughput analysis. It can be used at the expression level (cell lysates) or during the subsequent purification steps to monitor yield of specific protein. PMID- 12758266 TI - Polymerase chain reaction-based methods of DNA methylation analysis. AB - DNA methylation is the main epigenetic modification in humans, and changes in methylation patterns play an important role in tumorigenesis. Hypermethylation of normally unmethylated CpG islands in the promoter regions often occurs in important tumor suppressor genes, DNA repair genes, and metastasis inhibitor genes. The changes of methylation status of various gene promoters seem to be a common feature of malignant cells and these changes can occur early in the progression process. Therefore detection of aberrant promoter hypermethylation of cancer-related genes may be useful for cancer diagnosis or detection of cancer recurrence. The purpose of this review is to provide a summary of the most commonly used techniques for the study of DNA methylation. Current scientific literature involving methylation detection methods was reviewed with an emphasis on polymerase chain reaction (PCR)-based detection methods. The current methodologies may be broadly classed into PCR-based methylation assays and non PCR-based methylation assays. The problems and advantages of the different methods for detecting aberrant methylation are discussed. As the number of genes known to be hypermethylated in cancer is growing, the detection of aberrant promoter region methylation will be a promising approach for using DNA-based markers for the early detection of human cancers. Many techniques, especially PCR based methylation assay techniques, make it practical to use these new methylation biomarkers in early cancer diagnosis. PMID- 12758268 TI - A continuous fluorometric assay for inorganic pyrophosphate and for detecting single-base changes with polymerase chain reaction. PMID- 12758267 TI - Initial investigations into the ultrasonic lysis of microbial cells for the release of adenosine triphosphate. PMID- 12758269 TI - High-resolution separation of proteins by a three-dimensional sodium dodecyl sulfate polyacrylamide cube gel electrophoresis. PMID- 12758270 TI - A microarray method to evaluate the effect of CA mispairs on the accuracy of BstUI restriction endonuclease. PMID- 12758271 TI - Run parameters affecting protein patterns from second dimension electrophoresis gels. PMID- 12758272 TI - A protocol for separation and isolation of small and/or large DNA fragments with high yield using CL4B Sepharose. PMID- 12758273 TI - The Gram-negative bacterium Chlamydia trachomatis L2 stimulates tumor necrosis factor secretion by innate immune cells independently of its endotoxin. AB - The endotoxin of Chlamydia trachomatis L(2), the causative agent of lymphogranuloma venerum, has been described as an endotoxin with an atypical structure and weak stimulatory activity. It is, however, unclear whether chlamydial endotoxin plays a role in the stimulation of innate immune cells upon contact with the whole microorganism C. trachomatis L(2). We show here that chlamydial endotoxin and, as expected, Escherichia coli O55:B5 endotoxin depend on Toll-like receptor 4 without depending on Toll-like receptor 2 to stimulate bone marrow-derived dendritic cells to secrete tumor necrosis factor (TNF). In contrast, the whole microorganism C. trachomatis L(2) induces TNF secretion by innate immune cells independently of Toll-like receptor 4, while stimulation by E. coli O55:B5 depends on Toll-like receptor 4. Furthermore, although TNF secretion of the macrophage cell line RAW264.7 with chlamydial or E. coli O55:B5 endotoxin as well as with the bacterium E. coli O55:B5 is inhibited by the endotoxin-neutralizing compound polymyxin B, C. trachomatis L(2)-induced secretion of TNF cannot be reduced. In accordance with the literature, the potential of chlamydial endotoxin is more than 100-fold weaker than E. coli O55:B5 endotoxin on all cell types tested. We conclude that chlamydial endotoxin is unlikely to be involved in C. trachomatis L(2)-induced release of TNF by innate immune cells. PMID- 12758274 TI - Experimental transmission of Leishmania tropica to hamsters and mice by the bite of Phlebotomus sergenti. AB - Phlebotomus sergenti is a natural vector of Leishmania tropica. However, the ability of P. sergenti to transmit L. tropica by bite has not been proven experimentally yet. We have transmitted L. tropica to golden hamsters and BALB/c mice by the bite of P. sergenti. Sand flies and Leishmania both originated from an anthroponotic cutaneous leishmaniasis focus in Urfa, Turkey. P. sergenti females from a laboratory colony were infected by feeding on lesions of needle inoculated hamsters or mice. Gravid females were allowed to refeed on uninfected hosts 9-15 d after the infective feeding. At the second feeding, some infected females took a full blood meal, while others only a partial one; some females failed to feed at all. The ability of infected females to take a blood meal did not correlate with the parasite transmissibility. In four BALB/c mice, lesions developed after 1-6 months. In two albino hamsters (Mesocricetus auratus), lesions developed 1 month after the infective feeding, and Leishmania could be reisolated from these sites. Another hamster did not develop a lesion; however, the feeding site and the adjacent ear were PCR positive 1 year after infective feeding. Our results show that dissemination to other parts of host body occurs in L. tropica after sand fly bite. Experimental transmission of the parasite confirms that P. sergenti is a natural vector of L. tropica. PMID- 12758275 TI - Phenotypic and genotypic characterization of the neurovirulence and neuroinvasiveness of a large-plaque attenuated Japanese encephalitis virus isolate. AB - The virulent phenotypes of Japanese encephalitis virus (JEV) can be divided into neuroinvasiveness (NI) and neurovirulence (NV). In this study, two JEV antigenic variants, CH2195LA (large-plaque, attenuated) and CH2195SA (small-plaque, non attenuated), were passaged in suckling mice by intracerebral inoculation. Viruses at passage two and four were characterized in terms of NV and NI in weaning mice, as well as their in vitro growth characteristics in six cell lines. Following two brain-brain passages in mice, the attenuated variant CH2195LA was found to significantly restore the NV and NI by approximately 90% and 20-40%, respectively. The increased titers in THP-1 monocytic cells but not IMR-32 and Neuro-2A neuroblastoma cells were more correlated with the phenotypic changes of NI and NV in mice. Entire genomic sequencing was further performed to demonstrate that 14 nucleotides were altered in the attenuated variant CH2195LA following four brain-brain passages in mice, giving 12 amino acid changes, in prM-73, prM 80, E-161, E-170, E-276, NS2A-136, NS2A-215, NS3-346, NS4A-128, NS4B-196, NS4B 197, NS4B-198. This study indicated a cluster of amino acids which is involved in NV and NI of the JEV for mice and, perhaps, for humans. Elucidating the molecular basis of virulence of flaviviruses can provide valuable information for live attenuated vaccine development. PMID- 12758276 TI - Vibrio parahaemolyticus infectious for both humans and edible mollusk abalone. AB - The aims of this study are to report evidence of the first laboratory-acquired infection of Vibrio parahaemolyticus associated with handling experimentally infected abalones and to describe the virulence of the two bacterial strains tested in these animals. Two strains of V. parahaemolyticus, one from the stool of a patient with acute gastroenteritis (strain 880713) and the other from the hemolymph of a diseased small abalone Haliotis diversicolor supertexta (strain 880915), were identified and characterized. Both strains were lethal to small abalone, with similar LD(50) values (8.36-8.41 x 10(4) colony-forming units/g abalone). Laboratory-acquired infection resulted in one individual experiencing two episodes of acute gastroenteritis due to handling virulence tests during a 1 week interval. Our present results suggest that a V. parahaemolyticus strain isolated from the stool of a patient with gastroenteritis was infectious for small abalone, a major species of edible mollusk abalone cultured in Taiwan, while a similar strain isolated from hemolymph of a diseased small abalone was infectious for humans. This is the first report of V. parahaemolyticus virulent to small abalone as a zoonotic pathogen. PMID- 12758277 TI - A novel approach for collecting samples from fruit bats for isolation of infectious agents. AB - During the outbreak of Nipah virus encephalitis involving pigs and humans in peninsular Malaysia in 1998/1999, a conventional approach was initially undertaken to collect specimens from fruit bats by mist-netting and shooting, as an integral part of wildlife surveillance of the natural reservoir host of Nipah virus. This study describes a novel method of collecting fruit bats' urine samples using plastic sheets for isolation of Nipah virus. This novel approach resulted in the isolation of several other known and unidentified infectious agents besides Nipah virus. PMID- 12758279 TI - The role of natural killer T cells and other T cell subsets against infection by the pre-erythrocytic stages of malaria parasites. AB - T cells are critical mediators of immunity to the pre-erythrocytic stages of malaria parasites. In this review, we survey the role of the various T cell subsets in combating the pre-erythrocytic stages; in particular, the role of NK T cells. Moreover, we show how studies using malaria models have revealed a unique behavior of NK T cells, namely the bridging of innate and adaptive immunity. PMID- 12758278 TI - Vgamma2Vdelta2+ T cells and anti-microbial immune responses. AB - Vgamma2Vdelta2(+) T cells exist only in primates and constitute the majority of circulating human gammadelta T cells. Recent studies have demonstrated that this unique gammadelta T cell subpopulation can be a component of adaptive immune responses and contribute to anti-microbial immunity to infections. PMID- 12758280 TI - Early mechanisms of Leishmania infection in human blood. AB - In human blood, promastigotes bind natural antibodies and activate the classical complement pathway. C3-opsonized promastigotes immune-adhere within seconds to erythrocytes. Promastigote lysis by complement parallels C3 deposition kinetics, and ~90% of promastigotes are killed after 2.5 min. During infection, complement thus exerts strong selective pressure on Leishmania. Paradoxically, promastigote adaptation to the host immune adherence mechanism may provide the parasite a key to invasion. PMID- 12758281 TI - The critical role of adenosine A2A receptors in downregulation of inflammation and immunity in the pathogenesis of infectious diseases. AB - Adenosine can be described as a retaliatory metabolite, the production and release of which is usually enhanced under adverse environmental conditions. Binding via specific receptors, adenosine activates endogenous protective mechanisms aiming at the restoration of tissue homeostasis. While adenosinergic downregulation of tissue damage is beneficial in acute inflammation, chronic suppression of the immune system by adenosine may account for immunoparalysis in long-term septic patients. PMID- 12758282 TI - Responses to reactive oxygen intermediates and virulence of Salmonella typhimurium. AB - Salmonella typhimurium is an intracellular pathogen that can survive and replicate in macrophages. One of the host defense mechanisms that S. typhimurium encounters upon infection is superoxide produced by the phagocytes' NADPH oxidase. Salmonella has evolved numerous ways of coping with superoxide in the extracellular environment. In addition, Salmonella has to defend itself against superoxide produced as a by-product of aerobic respiration. Over the last decade, research on bacterial mutants has led to the identification of Salmonella strains that differ from their parental strain in susceptibility to superoxide in vitro. However, the consequences of such mutations for bacterial virulence are highly variable, indicating that superoxide sensitivity per se is not a characteristic that renders Salmonella less virulent. By discussing various bacterial mutants classified according to their in vitro sensitivity to superoxide, we will exemplify the complex mechanisms that Salmonella has evolved to cope with superoxide stress. PMID- 12758283 TI - Endophytes as sources of bioactive products. AB - An increase in the number of people in the world having health problems caused by various cancers, drug-resistant bacteria, parasitic protozoans, and fungi is a cause for alarm. An intensive search for newer and more effective agents to deal with these disease problems is now under way and endophytes are a novel source of potentially useful medicinal compounds. PMID- 12758284 TI - Lipid metabolism in Plasmodium falciparum-infected erythrocytes: possible new targets for malaria chemotherapy. AB - The emergence and spread of drug-resistant parasites coupled with the absence of an effective vaccine makes malaria treatment more complicated, and thus the development of new antimalarial drugs is one of the urgent tasks in malaria research. This review highlights lipid metabolism in Plasmodium parasite cells, the study of which would lead to providing new targets for therapeutic intervention. PMID- 12758285 TI - Louis Pasteur (1822-1895). AB - In Louis Pasteur's scientific career it is striking to note the exponential character of the research he introduced in all the fields he opened up. He offered fabulous opportunities to stereochemistry. He is acknowledged as one of the founders of microbiology. He established the possibility of anaerobic life. He pointed the way to epidemiology, public health, and the bacteriologic fight. He struggled against the idea of spontaneous generation of life. He irrevocably substantiated the microbial theory of infectious diseases. He demonstrated that bacterial virulence could be attenuated, he evidenced immunity and generalised the vaccination principle. He also was an incomparable experimenter. PMID- 12759093 TI - Maternal drug use in early pregnancy and infant cardiovascular defect. AB - The purpose of the paper is to identify maternal drug use that may be associated with an increased risk for cardiac defects in the offspring. A case-control study was performed with cases (cardiovascular defects without known chromosome anomalies) being identified from three Swedish health registers (n=5015) and controls being all infants born in Sweden during the period 1 July 1995-2001 (n=577,730). Information on drug exposure was obtained by interview in early pregnancy. Associations between maternal drug use and infant cardiovascular defect were identified for insulin, antihypertensives, fertility drugs, erythromycin, naproxen, anticonvulsants, nitrofurantoin, clomipramine, and budesonide in nasal preparations. Some of these associations are probably due to confounding from underlying disease or complaint, some may be due to multiple testing, some may be true drug effects. Further studies are needed to verify or reject these associations. PMID- 12759094 TI - Dibromochloropropane inhibits spermatogonial development in rats. AB - Exposure to the nematocide dibromochloropropane (DBCP) has caused prolonged oligo and azoospermia in men. There are questions regarding the cellular targets resulting in this effect. In this study we characterized an animal model, in which four daily injections of DBCP produced prolonged oligospermia in LBNF(1) rats without any indication of recovery. Between 6 and 20 weeks after DBCP treatment, 70% of seminiferous tubules showed an epithelium with Sertoli cells but no differentiating germ cells. About 20% of tubules contained differentiating germ cells and 10% showed occlusion or major morphologic alterations to Sertoli cells. Since gonadotropin levels and intratesticular testosterone (ITT) concentrations were elevated in the DBCP-treated rats, the failure of spermatogonial development could not have been a result of lack of these hormones. The tubules without differentiating germ cells contained actively proliferating and dividing type A spermatogonia, which underwent apoptosis instead of differentiation. Thus, the target for the damaging effect appears not to be the killing of stem spermatogonia, but the loss of their ability to undergo differentiation. The presence of type A spermatogonia in the atrophic tubules indicates the potential for intervention to restore spermatogenesis. PMID- 12759095 TI - In vivo exposure of female rats to toxicants may affect oocyte quality. AB - A potential endpoint for female reproductive toxicants is fertilizability of the oocytes. This endpoint has not been adequately examined for mammalian females. The objective of these studies was to evaluate fertilizability of rat oocytes following in vivo exposure to known male reproductive toxicants that exert effects via pathways that do not include endocrine disruption and to 4 vinylcyclohexene diepoxide, known to interfere with early follicular development. Oocytes were obtained from females following exposure and quality assessed by in vitro fertilization rate. One study evaluated fertilizability following 2 weeks exposure of females to inhaled tetrachloroethylene (2h/day, 5 days/week). The remaining studies evaluated fertilizability immediately following 2 weeks exposure via drinking water to tetrachloroethylene, trichloroethylene, the fuel oxidants methyl tertiary butyl ether (MTBE), ethyl tertiary butyl ether (ETBE), tertiary amyl methyl ether (TAME), and a metabolite of the first two ethers 2 methyl-1,2-propanediol (2M2P), and to 4-vinylcyclohexene diepoxide. The percentage of oocytes fertilized was reduced following inhalation exposure to tetrachloroethylene, or consumption of trichloroethylene or TAME. Fertilizability was not altered by exposures to the other reproductive toxicants or to the other fuel oxidants. Consistent with the reduced oocyte fertilizability following exposure to trichloroethylene, oocytes from exposed females had a reduced ability to bind sperm plasma membrane proteins. Female reproductive capability assessed by the endpoint, oocyte fertilizability, was reduced by exposure to trichloroethylene and inhaled tetrachloroethylene. PMID- 12759096 TI - Changes in reproductive organs of female rats treated with bisphenol A during the neonatal period. AB - The effects of neonatal administration of estrogenic agents on rat reproductive organs were examined. Either bisphenol A (BPA; 0.25, 1, or 4mg/pup) or 10 micro g 17beta-estradiol (E(2)) was given subcutaneously to Sprague-Dawley female rats during the neonatal period from post-natal day (PND) 0 to 9. Animals ovariectomized at 80 days were given subcutaneous injections of 1 micro g/kg E(2) for 3 days from PND 94 to 96. Clefts in the clitoris, early vaginal opening, irregular estrous cycles, a decrease in the area occupied by the corpora lutea (CL) in the ovary, and multiple cystic follicles in the ovary were found in the animals treated neonatally with 1mg BPA. Uterine fluid weight measured after E(2) treatment on PND 94-96 was less than controls. In addition to these abnormalities, unusual body weight gains, persistent vaginal cornification, and lack of CL were observed in females treated neonatally with 4mg BPA. The ovary weight on PND 80 and uterine fluid weight measured after E(2) treatment on PND 94 96 were less than controls for the 4mg BPA group. Neonatal treatment with 10 micro g E(2) induced similar abnormalities as found in the 4mg BPA group. These results show that BPA when given during the neonatal period caused changes in female reproductive organs. PMID- 12759097 TI - Multiple mechanisms are involved in apoptotic cell death in the mouse uterus and vagina after ovariectomy. AB - Withdrawal of sex hormones by gonadectomy results in rapid involution of mouse reproductive organs. To study the regression mechanism in the uterus and vagina after ovariectomy, histologic and biochemical changes were examined. Apoptotic cells were detected by in situ 3'-DNA nick end labeling method and electron microscopy, while the number of cells showing incorporation of bromo-deoxyuridine (BrdU) decreased in the uterus and vagina after ovariectomy. DNA fragmentation in the uterus was observed even at estrus and the degree of fragmentation increased after ovariectomy. DNA fragmentation in the vagina occurred 1-5 days after ovariectomy. Semi-quantitative RT-PCR revealed that expression of Fas-ligand and tumor necrosis factor-alpha (TNF-alpha) mRNA in the uterus and vagina was increased by ovariectomy. These results suggest that apoptotic cell death is induced by ovariectomy through the mediation of both Fas and TNF-alpha in the mouse uterus and vagina; however, uterine and vaginal cells in CBA lpr(cg)/lpr(cg) mice lacking functional Fas showed apoptosis, indicating that Fas is not the sole regulator of apoptosis in female reproductive organs in mice. PMID- 12759098 TI - Effects of lindane on oocyte maturation and preimplantation embryonic development in the mouse. AB - Lindane, an organochlorine insecticide, is suspected of preimplantation embryonic toxicity based on in vitro experiments with bovine and murine embryos. To verify this hypothesis in vivo we tested lindane for developmental alterations during early embryonic cleavage in the mouse. Two treatment schedules were tested: three daily doses of 15 or 25mg/kg b.w. lindane were orally administered to female mice either before mating or immediately after mating. Morphologic alterations (lysis or fragmentation of blastomeres, developmental arrest) of two-cell embryos and morulae were evaluated by inverted microscopy. In addition, cytologic abnormalities and cell proliferation delay, possibly induced during the first four cleavage cycles, were evaluated by fluorescent microscope analysis of the number and morphology of blastomere nuclei. A statistically significant increase of degenerating two-cell embryos was induced by exposure of preovulatory oocytes to the highest tested lindane dose. Early cleavage embryos exposed to the same dose showed a lower average number of blastomeres per morula, as well as a 40% reduction of the mitotic index with respect to matched controls. However, mean values in individual litters were variable and litter analysis did not show a lindane-related effect. One possible mechanism for the observed effects could be the recently demonstrated inhibitory action of lindane on gap junction-mediated cell communication between oocyte and cumulus cells. A comparison between human exposure levels and experimental doses based on measured and predicted blood concentrations suggests that there are ample margins of safety for human embryonic development at the present exposure levels. PMID- 12759099 TI - Effect of mono-(2-ethylhexyl) phthalate on bovine oocyte maturation in vitro. AB - The effect of mono(2-ethylhexyl) phthalate (MEHP) on bovine oocyte maturation in vitro was examined. Cumulus-oocyte complexes (COCs) were cultured in maturation medium supplemented with various levels of MEHP for 24h, and then examined for the degree of cumulus expansion and the stage of maturation. A higher percentage of oocytes remained at the germinal vesicle (GV) stage after exposure to 75 and 100 micro M MEHP treatments (13.8 and 44.9% of oocytes, respectively) than the control (2.1% of oocytes). The proportion of oocytes that progressed to the metaphase II (MII) stage was significantly decreased with 25 micro M (59.6% of oocytes), 50 micro M (19.8%), 75 micro M (21.3%), and 100 micro M (3.1%) treatments than the control (77.3%). MEHP did not affect the process of cumulus expansion. For denuded oocytes, MEHP treatment of 50-100 micro M resulted in a significantly higher rate of oocytes remained at the GV stage compared to the control (53.4, 80.2, 88.4, and 5.4%, respectively). The rate of MII formation was significantly decreased with 10 micro M (60.9%) and 25 micro M (22.5%) MEHP treatments compared to control (68.9%). Furthermore, with 50, 75 or 100 micro M MEHP, no oocyte reached the MII stage. When COCs were cultured for 24h with 50 or 100 micro M MEHP and then cultured for an additional 24h in MEHP-free medium, most of the oocytes reached the MII stage (71.1 and 64.5%, respectively).Taken together, these results indicate that MEHP, at doses lower than those reported in blood transfusion patients, could negatively modulate bovine oocyte meiotic maturation in vitro, suggesting possible risks for human and other mammalians reproductive health. PMID- 12759100 TI - Novel Calbindin-D(9k) protein as a useful biomarker for environmental estrogenic compounds in the uterus of immature rats. AB - The compounds that bind the estrogen receptors (ER) and induce or modulate an ER mediated response can be defined as estrogenic endocrine disruptors (EDs). We demonstrated that environmental estrogens induced the uterine CaBP-9k mRNA in rats using Northern blot assay suggesting that CaBP-9k gene could be a biomarker for the estrogenicity of chemicals in the previous studies. In the present studies, we further collaborated this idea by investigating the regulation and localization of CaBP-9k protein in response to estrogenic compounds. Immature rats were injected with estrogenic chemicals, 4-tert-octylphenol (OP), nonylphenol (NP) and bisphenol A (BPA) or 17beta-estradiol (E2). After treatment with these estrogenic compounds, the effects on the accumulation of CaBP-9k protein and uterine localization were examined by Western blot analysis and immunohistochemical staining (IHC), respectively. A dose- and time-dependent increase in CaBP-9k protein was observed in the uterus of immature rats when treated with OP and NP. In addition, treatment with BPA resulted in a significant increase in CaBP-9k protein at dose of 500mg/kg BW/day. Taken together, CaBP-9k protein is strongly up-regulated by estrogenic compounds (OP, NP and BPA) and E2 itself in the uterus of immature rats. These results indicate that CaBP-9k protein can be a useful biomarker for detection of the estrogenicity of putative estrogenic compounds. Thus, regarding to risk assessment, we propose that CaBP-9k protein assay in the immature rat uterus can be a very sensitive and powerful tool to identify compounds with estrogenic activity when used in combination with classical assays. PMID- 12759101 TI - Nonthermal effects of mobile-phone frequency microwaves on uteroplacental functions in pregnant rats. AB - Exposure to high-density microwaves can cause detrimental effects on the testis, eye, and other tissues, and induce significant biologic changes through thermal actions. To examine nonthermal effect of continuous wave (CW) 915MHz microwaves used in cellular phones, we compared the effects of microwaves with those of heat. Thirty-six pregnant rats were assigned to six groups: rats exposed to microwaves at 0.6 or 3mW/cm(2) incident power density at 915MHz for 90min, rats immersed in water at 38 or 40 degrees C, which induces about the same increase in colonic temperature of 1.0 or 3.5 degrees C as 0.6 or 3mW/cm(2) microwaves, respectively; rats immersed in water at 34 degrees C, which is considered to be thermoneutral; and control rats. We identified significant differences in the uteroplacental circulation, and in placental endocrine and immune functions between pregnant rats immersed in water at 34 and 38 degrees C, but not between rats immersed at 38 degrees C and those exposed to microwaves at 0.6mW/cm(2). By contrast, we observed significant decreases in uteroplacental blood flow and estradiol in rats exposed to microwaves at 3mW/cm(2) as compared with those immersed in water at 40 degrees C. These results suggest microwaves at 0.6mW/cm(2) at 915MHz, equal to a specific absorption rate (SAR) of 0.4W/kg, which is the maximum permissible exposure level recommended by the American National Standards Institute (ANSI), do not exert nonthermal effects on blood estradiol and progesterone, on splenic natural killer cell activity, on the uteroplacental circulation. PMID- 12759102 TI - Two-generation reproductive study in mink fed diisopropyl methylphosphonate (DIMP). AB - Two generations of "Ranch Wild" mink (Mustela vison) were fed the organophosphate diisopropyl methylphosphonate (DIMP) at 0, 150, 450, or 1250ppm, to determine potential toxicity to the dams. Chemical, hematologic, necropsy, and microscopic examinations were performed on all parental animals and representative kits. The F0 and F1 dams had 3.4 and 4.6% mortality, respectively, distributed among all groups and not attributed to DIMP exposure. Adverse effects were mild and limited to the highest dose group. Plasma cholinesterase was reduced 40% (F0) and 31% (F1), as was whole blood cholinesterase (16 and 8.5%). Heinz bodies were present in 2.8% (F0) and 1.3% (F1) of erythrocytes. The erythrocyte count was reduced 6.3% in the F0. Reproductive efficiency was not affected. The mink were not uniquely susceptible to DIMP, relative to the literature on other species. The no observed adverse effect level (NOAEL), based on the 450ppm group of F1 females, was 56.5mg DIMP/kgBW per day; the lowest observed adverse effect level (LOAEL) was 329.5mg DIMP/kgBW per day. PMID- 12759103 TI - Evaluation of an in utero through lactational exposure protocol for detection of estrogenic effects of ethinyl estradiol on the offspring of rats: preliminary trial. AB - As a preliminary trial of an in utero through lactational exposure protocol, ethinyl estradiol, 0, 0.5, 5, or 50 micro g/kg/day, was administered by gavage to pregnant Crj: CD (SD) IGS BR rats from gestational day (GD) 7 to day 18 after delivery to evaluate the efficacy of this protocol and to estimate optimal endpoints. The dams showed no abnormalities. Cleft phallus was observed in female offspring at 50 micro g/kg. Other than a retardation of body weight gain in both sexes at 50 micro g/kg/day, no other abnormal findings were detected. The fact that cleft phallus was the only change induced suggests that the protocol is applicable to the detection of effects of estrogenic chemicals given to pregnant rats on offspring development and that the morphology of the female external genitalia may be a useful endpoint. PMID- 12759104 TI - Offspring sex ratio is not associated with parental GSTT1 and GSTM1 null genotypes. AB - The present study evaluated the association between offspring sex ratio and parental inherited variations of xenobiotic metabolism by glutathione S transferase (GST) T1 and GSTM1. Polymorphisms of GSTT1 and GSTM1 were investigated in 235 Iranian Muslims (153 males, 82 females) using polymerase chain reaction-based method. Statistical analysis of 890 offspring in the study families revealed that there was no association between GSTM1 null genotype of either the fathers (OR=1.28; 95% CI=0.88-1.85; P=0.17) or mothers (OR=0.77; 95% CI=0.49-1.19; P=0.22) and offspring sex ratio. There was no association between paternal GSTT1 null genotype and sex ratio in offspring (OR=1.04; 95% CI=0.71 1.54; P=0.84), whereas there was a weak association between maternal GSTT1 null genotype and offspring sex ratio (OR=1.54; 95% CI=1.0-2.38; P=0.05). Combination of parental GSTT1 and GSTM1 genotypes showed no association with offspring sex ratio. PMID- 12759105 TI - Methanol metabolism and embryotoxicity in rat and mouse conceptuses: comparisons of alcohol dehydrogenase (ADH1), formaldehyde dehydrogenase (ADH3), and catalase. AB - Mouse embryos are more sensitive than rat embryos in response to methanol (CH(3)OH) and its ability to elicit developmental abnormalities. Intrinsic differences in the metabolism of CH(3)OH to formaldehyde (HCHO) and formic acid (HCOOH) by the enzymes alcohol dehydrogenase (ADH1), formaldehyde dehydrogenase (ADH3), and catalase may contribute to the observed species sensitivity. Specific activities for enzymes involved in CH(3)OH metabolism were determined in rat and mouse conceptuses during the organogenesis period of 8-25 somites. Spatial activity relationships were also compared separately in heads, hearts, trunks, and the visceral yolk sac (VYS) from early (7-12 somites) and late (20-22 somites) organogenesis-stage rat and mouse embryos. Catalase activities were similar between rat and mouse conceptuses. In the mouse heart, catalase activities were consistently lower when compared to other tissues. Specific activities for catalase were consistently highest in the VYS of both species when compared to other tissues of the embryo. These activities were highly significant in the 6-12 somite VYS. ADH1 activities were significantly higher in embryos when compared to VYS in both species, except for a 27% lower activity in the early 8 10 somite mouse embryo. Mouse ADH1 activities in the VYS were significantly lower throughout the organogenesis period when compared to the rat VYS or embryos of either species. Mouse activities were lower overall in specific tissues of the embryo but maintained the same relative proportions as in the rat. ADH3 activities in the rat VYS were significantly higher by 20% than those in the mouse. Mouse embryo ADH3 activities were slow to mature, starting at a level 42% below rat, and failed to reach optimal levels until the 14-16-somite stage. Heart ADH3 activities were also significantly lower in the mouse embryo at the 7-12 somite stage. Both species have lower ADH3 activities in the early heart, relative to other embryonic tissues. These results show a more slowly maturing capacity of the mouse embryo to remove HCHO, which provides a rationale for increased sensitivity of this species to CH(3)OH-induced embryotoxicity and teratogenicity. PMID- 12759107 TI - Lack of effect of 5HT3 antagonist in mediating subjective and behavioral responses to cotinine. AB - Previous studies have shown that cotinine, a metabolite of nicotine, may antagonize some of the therapeutic effects of nicotine. The mechanisms underlying cotinine's effects are unclear, but cotinine has been observed to increase serotonin levels in the brain. Thus, it is possible that blocking serotonin effects may antagonize the actions of cotinine, thereby reducing its impact on responses to nicotine. This study determined whether granisetron, a 5HT(3) receptor antagonist, would enhance the efficacy of the nicotine patch. Subjects were randomly assigned to one of the three granisetron conditions (N=43 for 2 mg/day; N=43 for 1 mg/day; N=42 for 0 mg/day) and asked to take the assigned medication daily during 15 days of tobacco abstinence. Because we were interested in interactions between cotinine and serotonin, all groups were also treated with a 21-mg nicotine patch. Assessments of withdrawal symptoms were made for 1 week during baseline smoking and several times during the experimental period. There was a near but nonsignificant difference among groups on a measure of tobacco withdrawal and no significant differences on global measures of drug effects or physiological measures. The data do not strongly support the hypothesis that 5HT(3) agonism is the mechanism by which cotinine offsets the effects of nicotine. PMID- 12759108 TI - Effects of dopamine antagonists on the timing of two intervals. AB - Rats were trained on a two-interval (12 and 36 s) temporal production task (the peak procedure). Test sessions were conducted in which either the D(1) antagonist SCH-23390 (SCH; 0.02, 0.04, 0.06 mg/kg) or the D(2) antagonist haloperidol (HAL; 0.05, 0.1, 0.2 mg/kg) were injected prior to testing. Both drugs affected the amount of responding, but only HAL affected timing. Under HAL, both intervals were overestimated, consistent with a HAL-induced decrease in clock speed. Drug induced decreases in response output were more profound for the long interval than the short. In addition, there was evidence of HAL- and SCH-induced delays in response initiation that were more severe for the long interval, perhaps owing to its status as a weaker conditioned stimulus. PMID- 12759109 TI - Differential vulnerability to motor deficits in second replicate HAS and LAS rats following neonatal alcohol exposure. AB - Children exposed prenatally to alcohol suffer from a variety of behavioral alterations. However, variation exists in the pattern and severity of these alcohol-related neurodevelopmental disorders. We examined the influence of alcohol sensitivity in the etiology of fetal alcohol effects by studying rat lines selectively bred for extremes in alcohol-induced sleep time: high-alcohol sensitive (HAS) and low-alcohol-sensitive (LAS) rats. Using subjects from the first replicate, we previously reported that HAS rats exposed to alcohol during development were more vulnerable to ethanol-induced hyperactivity and motor deficits compared to LAS rats. To determine if these effects were, in fact, related to the trait for which these subjects were selected, the present study examined the consequences of developmental alcohol exposure in second replicate HAS and LAS rats. Second replicate HAS and LAS rats, as well as Sprague-Dawley rats, were exposed to 6.0 g/kg/day ethanol on Postnatal Days (PD) 4-9, a period of brain development equivalent to the third trimester, via an artificial rearing procedure. Artificially and normally reared controls were included. Activity was measured on PD 18-21 and parallel bar motor coordination on PD 30-32. Ethanol exposure produced hyperactivity in all genetic groups, and there were no differences among HAS and LAS rats. In contrast, consistent with findings from the first replicate, ethanol-exposed HAS rats were more impaired on the motor coordination task compared with LAS rats. These data suggest that genetically mediated responses to alcohol may relate to behavioral vulnerability to motor deficits following developmental alcohol exposure. They also provide evidence that genetic factors play a role in fetal alcohol effects and suggest that phenotypic markers may indicate individuals at high risk for some fetal alcohol effects. PMID- 12759110 TI - Blockade of histamine H2 receptors of the periaqueductal gray and inferior colliculus induces fear-like behaviors. AB - Electrical and chemical stimulation of the dorsal periaqueductal gray matter (dPAG) and the inferior colliculus (IC) induces escape behavior, usually accompanied by autonomic responses and antinociception. Recently, we presented evidence for a tonic inhibitory control exerted by H(2) histamine receptors on defensive behaviors generated in these midbrain tectum sites. Since treatments of these areas that elicit the defensive behavior repertoire frequently also have anxiogenic effects, we here used the elevated plus-maze (EPM) test for assessing the effects of microinjections of histamine (5-40 nmol), dimaprit (5-10 nmol) and ranitidine (10-30 nmol) into either dPAG or IC, which have a relative abundance of histamine-containing cells and histaminergic receptors. Dimaprit is an agonist and ranitidine is an antagonist of H(2) histamine receptors. Immediately after the injections, the animals were submitted to the EPM test. Whereas dPAG injections of dimaprit had no behavioral effects, histamine (40 nmol) caused a significant reduction in exploratory activity. On the other hand, ranitidine alone or following saline had aversive-like effects in both structures, i.e. reduced open arm, but not closed arm, entries. This pattern is usually interpreted as representing an anxiogenic effect. These effects were more pronounced after injection into dPAG than into IC. Freezing, the most prominent effect produced by ranitidine, was significantly inhibited by histamine as well as dimaprit. Thus, H(2) receptor blockade has fear-like action in the midbrain tectum with predominance in the dPAG. Such an action can be understood as a concomitant of defensive behavior, which has been shown to be a consequence of H(2) receptor antagonism in both dPAG and IC. The functional significance of the different effects of H(2) receptor blockade in dPAG and IC is discussed in the light of the probable distinct roles of these structures in the organization of defensive behavior. PMID- 12759111 TI - Dose-response effects of chronic lithium regimens on spatial memory in the black molly fish. AB - Lithium is widely used in the management of bipolar disorder, yet memory impairment is a serious side effect. To assess the effects of lithium on spatial working and reference memories, we have employed a plus maze utilizing spontaneous alternation (SA) and place-learning paradigms in two experiments with the black molly fish. Four treatment groups were gavaged with 20 microl of a 10, 100, or 1000 mM lithium chloride (LiCl) solution or ddH(2)O vehicle every 12 h for 22 to 24 days. On Day 15, subjects began an 8-day SA task or a 10-day place learning task. Results indicate that there is a significant difference in SA performance among the treatment groups for Days 1, 2, and 3. Results of the place learning task indicate that the 1 M dose group needed significantly more trials to reach criterion and made significantly fewer correct first choices than the other dose groups. Capillary ion analysis determinations of plasma and brain lithium levels illustrate linear dose-response relationships to doses administered. Regression analyses indicate that there is a relationship between SA performance and plasma/brain lithium levels during the initial part of testing. Collectively, the results indicate that chronic lithium administration impairs spatial working and reference memories. PMID- 12759112 TI - The relationship between cocaine-induced increases in NAC1 and behavioral sensitization. AB - Repeated exposure to cocaine can cause long-term behavioral changes in mammals, including an augmented locomotor response known as behavioral sensitization. A major goal of research is the identification of molecules associated with these behaviors. NAC1, a member of the POZ/BTB transcription factor family, exhibited increased mRNA levels in the nucleus accumbens of the rat weeks after cocaine use. NAC1 exists as two isoforms, each demonstrating the ability to inhibit transcription, but to different extents. The present experiments examined the time course for both NAC1 isoforms after five consecutive days of systemic cocaine administration in male rats. Tissues were collected from several central nervous system regions and underwent Western blot analysis. There was significantly greater expression of the long isoform, lNAC1 (cocaine 1.341+/ 0.641; saline 1+/-0.321; P=.044), and the short isoform, sNAC1 (cocaine 3.038+/ 2.816; saline 1+/-0.720; P=.001), in the nucleus accumbens of cocaine-treated rats. The olfactory tubercle also showed a significant increase, but only in sNAC1 expression and at only one time period. No other significant differences were observed for either isoform of NAC1 in any other brain region. The expression of lNAC1 exhibited an inverse relationship with behavioral sensitization in rats 1-3 months following repeated cocaine injections predicting approximately 40% of the variance in the behavior variables (R(2)=.387; and P=.031 for distance and P=.025 for ambulatory count). These results indicate that NAC1 expression is increased for a period of several months after chronic cocaine exposure. Furthermore, these data suggest that NAC1 may function as an endogenous inhibitor of behavioral sensitization. NAC1 represents a target for future studies examining cocaine-induced behavioral changes. PMID- 12759113 TI - Dopamine D1 and D2 antagonists reduce the acquisition and expression of flavor preferences conditioned by fructose in rats. AB - The effects of dopamine (DA) D(1) and D(2) receptor antagonists on the acquisition and expression of flavor-preferences conditioned by the sweet taste of fructose were examined. Food-restricted rats were trained over eight alternating one-bottle sessions to drink an 8% fructose solution containing one novel flavor (CS+) and a less preferred 0.2% saccharin solution containing a different flavor (CS-). Three groups of rats were treated daily with either vehicle (control group), SCH23390 (200 nmol/kg; D(1) group), or raclopride (200 nmol/kg; D(2) group) during training. Additional groups of vehicle-treated rats had their daily training intakes matched to that of the D(1) and D(2) groups. Preferences were assessed in two-bottle tests with the CS+ and CS- flavors presented in 0.2% saccharin solutions following doses of 0, 50, 200, 400, or 800 nmol/kg of either D(1) or D(2) antagonists. The D(1) and D(2) groups, unlike the control and yoked-control groups, failed to display a significant CS+ preference in the two-bottle tests following vehicle treatment. In addition, treatment with SCH23390 prior to the two-bottle tests blocked the expression of the CS+ preference in the control groups. Pretest raclopride treatment attenuated the CS+ preference at some dose levels. Raclopride also attenuated the preference for fructose in rats given two-bottle training with the CS+/fructose (CS+/F) and CS /saccharin (CS-/S) solutions. These findings indicate that D(1) and D(2) antagonists block flavor-preference conditioning by sweet taste and that D(1), and to a lesser extent D(2), receptor antagonists attenuate the expression of a previously acquired preference. PMID- 12759114 TI - Methylphenidate-induced motor activity in rats: modulation by melatonin and vasopressin. AB - Methylphenidate (MPH), a dopamine (DA) reuptake inhibitor, is well known to enhance motor activity, in part depending on the time of its application during the light-dark cycle. Moreover, after MPH administration, the hypothalamo neurohypophysial axis including the neuropeptide vasopressin (AVP) was found influenced. Both the latter and behavioural effects of central AVP can also be modulated by the pineal gland with its light-dark-dependent activity. The present study was performed to investigate whether the pineal gland, its hormone melatonin (Mel), and AVP are involved in the MPH-evoked stimulation of activity. After application of 10 mg/kg MPH, the motor activity in pinealectomised (PE) rats was significantly higher than in sham-operated (SO) animals. After application of 250 microg Mel before MPH treatment, the stimulation of motor activity was diminished in PE rats and augmented in SO animals; however, when SO and PE rats were compared after Mel pretreatment, the reaction to MPH was nearly identical. Blocking the endogenous AVP by 25 or 1 microg of the V1a receptor antagonist d(CH(2))(5)[Tyr(Me)(2)]AVP (AAVP) before MPH treatment significantly augmented the motor activity in SO rats only and abolished the differences seen between SO and PE animals after MPH application. The present results indicate that the behavioural stimulation of MPH was modulated by both the pineal gland with its hormone Mel as well as the neuropeptide AVP. PMID- 12759115 TI - The effects of alprazolam on conditioned place preferences produced by intravenous heroin. AB - Case studies reveal that opiate addicts often self-medicate with benzodiazepine (BDZ) tranquilizers prior to taking their opiate. Our laboratory has previously utilized the conditioned place preference paradigm to confirm that BDZs can augment the affective response to heroin in laboratory animals. The combination of alprazolam and varying doses of intravenous heroin resulted in a leftward shift of the heroin dose-response curve. The present experiment was devised to extend the previous findings by examining the ability of varying alprazolam doses (0.125, 0.25, or 0.5 mg/kg ip) to potentiate the reward of a single challenge dose of heroin (0.025 mg/kg iv). The results demonstrate that a nonrewarding dose of alprazolam (0.125 mg/kg) and intravenous heroin can interact to produce reliable place preferences. The data thereby support prior work from our laboratory regarding the synergistic actions of BDZs and opiates. PMID- 12759116 TI - Noradrenergic and serotonergic blockade inhibits BDNF mRNA activation following exercise and antidepressant. AB - Antidepressants and physical exercise have been shown to increase the transcription of hippocampal brain-derived neurotrophic factor (BDNF). Much evidence regarding the initial actions of antidepressant medications as well as exercise leads to the hypothesis that noradrenergic (NE) and/or serotonergic (5 HT) activation is a key element in the BDNF transcriptional elevation common to both interventions. Currently, we used short-term beta-adrenergic, 5-HT(1A), or 5 HT(2A/C) receptor blockade to characterize the influence of NE and 5-HT systems on BDNF transcription during physical exercise and antidepressant treatment. In situ hybridization revealed that beta-adrenergic blockade significantly blunted the BDNF mRNA elevations due to exercise, and also inhibited the modest elevations in the CA3 and dentate gyrus following short-term treatment with tranylcypromine. In contrast, 5-HT(2A/C) blockade only minimally altered exercise induced BDNF mRNA levels, but inhibited up-regulation of BDNF transcription via tranylcypromine. Finally, 5-HT(1A) blockade did not inhibit exercise-induced BDNF mRNA elevations, but significantly enhanced levels above those achieved with exercise alone in the CA4. These results suggest that NE activation via beta adrenergic receptors may be essential for both exercise and antidepressant induced BDNF regulation. 5-HT(1A) and 5-HT(2A/C) activation, on the other hand, appear to be most important for antidepressant-induced BDNF regulation, but may also participate significantly in exercise-induced regulation in the CA4. PMID- 12759117 TI - Moderate doses of ethanol partially reverse avoidance learning deficits in high alcohol-drinking rats. AB - We previously reported that ethanol-naive high-alcohol-drinking (HAD1 and HAD2) rats exhibited selective deficits in active avoidance learning, as compared to low-alcohol-drinking (LAD1 and LAD2) rats, in a signaled bar-pressing task [Alcohol. Clin. Exp. Res. 24 (2000) 1778]. In the current study, we used appetitive and aversive learning tasks to assess whether administration of ethanol influences approach and avoidance learning in HAD and LAD rats. Rats were administered 0.0, 0.5, 1.0, or 1.5 g ethanol/kg body weight during appetitive and aversive conditioning sessions. We found that ethanol impaired acquisition of the appetitive conditioned response in a dose-dependent manner in both HAD and LAD rats, with 1.5 g/kg ethanol producing the greatest deficits. Notably, moderate doses of ethanol (0.5 and 1.0 g/kg) partially reversed avoidance learning deficits in HAD rats, but only when appetitive conditioning preceded aversive conditioning. The highest dose (1.5 g/kg EtOH) abolished avoidance responding altogether in HAD rats. Avoidance responding in LAD rats was not affected by any dose of ethanol. These results are consistent with previous studies suggesting that alcohol preference may be associated with increased fear or anxiety, but the conditions under which ethanol produces a reduction of fear and anxiety in HAD rats appear to be relatively complex. PMID- 12759118 TI - Effects of preexposure to dexfenfluramine, phentermine, dexfenfluramine phentermine, or fluoxetine on sibutramine-induced hypophagia in the adult rat. AB - The antiobesity drug sibutramine suppresses food intake via inhibition of reuptake of both norepinephrine (NE) and serotonin (5-HT) into brain terminals. The present study examined whether preexposure to other antiobesity drugs (fluoxetine [FLUOX], phentermine [PHEN], and dexfenfluramine [DEX]) that alter noradrenergic and/or serotonergic activity in brain induces tolerance or sensitization to the subsequent hypophagic action of sibutramine. Accordingly, adult male rats were treated (administered orally once per day for 21 days) with DEX (0, 1, or 3 mg/kg) and/or PHEN (0, 5, or 10 mg/kg), alone and in combination, or with the selective 5-HT reuptake inhibitor FLUOX (0, 15, or 30 mg/kg). Daily administration of PHEN persistently reduced food intake and body weight whereas tolerance developed to the hypophagic action of DEX or of FLUOX within the first week of daily administration. Moreover, low doses of DEX (1 mg/kg) and PHEN (5 mg/kg) interacted in a supra-additive manner to inhibit food intake and water intake and decrease body weight over the 21-day exposure period. After a recovery period of 9 days, a series of food intake trials were conducted to assess the hypophagic action of sibutramine (0, 1, 3, and 9 mg/kg po). Preexposure to PHEN (5 or 10 mg/kg), DEX (3 mg/kg), or FLUOX (30 mg/kg) resulted in a significant attenuation of the hypophagia induced by sibutramine over an 8-h, but not a 2-h, testing period. The pattern of cross-tolerance noted in this study is consistent with the observation that sibutramine inhibits eating via an interaction with noradrenergic and serotonergic mechanisms. Whether PHEN and DEX preexposure in humans alters subsequent sibutramine effectiveness is unknown. PMID- 12759119 TI - Differential effects of intraplantar capsazepine and ruthenium red on capsaicin induced desensitization in mice. AB - Intraplantar injection of capsaicin (1.6 microg/paw) into the mouse hindpaw produced an acute paw-licking/biting response. This study was designed (1) to investigate the antinociceptive effects of intraplantar administration of capsazepine, a competitive vanilloid receptor antagonist, and ruthenium red, a noncompetitive antagonist, in the nociceptive licking/biting response induced by intraplantar injection of capsaicin, and (2) to determine whether these compounds were able to prevent capsaicin-induced desensitization in mice. Both capsazepine and ruthenium red produced a dose-dependent reduction in the capsaicin-induced nociceptive response. In licking/biting response to intraplantar capsaicin, ruthenium red was more potent than capsazepine in producing antinociceptive activity as assayed by the capsaicin test. The first injection of capsaicin induced a profound desensitization to the second and third injections of capsaicin at the interval of 15 or 30 min. The capsaicin-induced desensitization was prevented dose-dependently by antinociceptive doses of capsazepine, whereas ruthenium red in doses exhibiting antinociceptive activity was without effect on capsaicin-induced desensitization. The present results suggest that both capsazepine and ruthenium red can produce a local peripheral antinociceptive action, which may be mediated by inhibiting the membrane ion channel activated by capsaicin. In addition, these data suggest that capsazepine may act in the mechanism clearly different from ruthenium red in the capsaicin-induced nociceptive desensitization. PMID- 12759120 TI - Locomotion, stereotypy, and dopamine D1 receptors after chronic "binge" cocaine in C57BL/6J and 129/J mice. AB - We have shown that C57BL/6J and 129/J mice differ in their behavioral response to "binge" pattern cocaine (three daily injections of 15 mg/kg separated by 1 h). To determine if these differences persist during chronic binge cocaine administration, we examined the effects of 14-day binge pattern cocaine on home cage behavior. Since the dopamine D(1) receptor may be an important mediator of cocaine-induced locomotor activity, we examined binding to the dopamine D(1) receptor. Locomotor activity was increased by chronic binge cocaine in C57BL/6J (P<.0001) but not in 129/J mice. C57BL/6J mice developed tolerance to the locomotor-activating effects of cocaine. Stereotypic responses were greater in C57BL/6J than in 129/J mice (P=.03), with neither tolerance nor sensitization in either strain. Dopamine D(1) receptor binding in the nucleus accumbens and olfactory tubercle did not differ between strains and was not affected by chronic binge cocaine. In the caudate putamen, subregion specific strain differences in dopamine D(1) receptor binding were observed; chronic binge cocaine increased dopamine D(1) receptor binding in the caudal (P<.05), but not rostral caudate putamen. There was no correlation between locomotor activity or stereotypy and dopamine D(1) receptor density. Thus, with chronic binge cocaine administration, behavioral differences persist between the C57BL/6J and 129/J mice, and cocaine induced locomotor activity is not correlated with changes in dopamine D(1) receptor binding. PMID- 12759121 TI - Chronic treatment with antipsychotics in rats as a model for antipsychotic induced weight gain in human. AB - Several clinical reports have demonstrated that most antipsychotics of the new generation, but not the typical antipsychotic haloperidol, induce weight gain in schizophrenic patients. Since weight gain induces serious health complications in humans, it is crucial to test upcoming antipsychotic compounds in an animal model of weight gain. With the aim of evaluating whether the rat can be used as a model for antipsychotic-induced weight gain, we have investigated the effect of chronic treatment (3 weeks) with one antipsychotic drug inducing weight gain in clinic (olanzapine) and one antipsychotic not inducing weight gain in clinic (haloperidol), on food and water intake and body weight gain in rats. We included both female and male rats in this study. To reduce spontaneous high food intake in rats, and to be able to evaluate the treatment effect on a potential increase of food intake or metabolic changes, we allowed animal to receive only low palatability chow. In male rats, none of the two compounds induced weight gain, but in female rats, both compounds induced weight gain. Consequently, the effect observed in rats does not match the clinical situation, and Wistar rats in this set-up cannot be considered a relevant model for antipsychotic-induced weight gain in humans. PMID- 12759122 TI - Time-dependent effects of cycloheximide on long-term memory in the cuttlefish. AB - When shown prawns in a glass tube, cuttlefish promptly learn to inhibit their predatory behavior and retain this ability for a long time. The cellular and molecular mechanisms of this long-term memory (LTM) are not yet known. In this study, we analyzed the dependency of LTM on de novo brain protein synthesis. Cycloheximide (CXM), a protein synthesis inhibitor, is injected intravenously immediately, 1 h, 3 h, 4 h or 6 h after the training. Retention is tested 24 h posttraining. The injections of CXM revealed one period of memory sensitivity to pharmacological intervention. CXM administered immediately or 6 h after training has no effect on LTM. Conversely, injections given between 1 and 4 h posttraining resulted in amnesia. Taken together, findings of this study establish for the first time in Sepia officinalis that de novo protein synthesis is an essential and time-dependent event for LTM formation of this form of associative learning. PMID- 12759123 TI - Presynaptic regulation of neurotransmitter release in the cortex of aged rats with differential memory impairments. AB - Cluster analysis of water-maze reference-memory performances of 25-27-month-old (compared to 3-5-month-old) rats distinguished subpopulations of young adult rats (YOUNG), aged rats with no significant impairment (AU), aged rats with moderate impairment (AMI), and aged rats with severe impairment (ASI). In the frontoparietal cortex, we subsequently assessed the electrically evoked release of tritium in slices preloaded with [3H]choline, [3H]noradrenaline (NA), or [3H]serotonin (5-HT) and the effects of an agonist (oxotremorine, UK 14,304, and CP 93,129) of the respective autoreceptors. Cholinergic and monoaminergic markers were measured in homogenates. Overall, aged rats exhibited reduced accumulation of [3H]choline (-25%) and weaker evoked transmitter release (in % of accumulated tritium: -44%, -20%, and -34%, for [3H]acetylcholine, [3H]NA, and [3H]5-HT, respectively). In all rats, the inhibitory effects of the autoreceptor agonists on the evoked release of [3H] were comparable. Acetylcholinesterase (AChE), not choline acetyltransferase (ChAT), activity was reduced. The results suggest age related modifications in the cholinergic, noradrenergic, and serotonergic innervation of the frontoparietal cortex, alterations of evoked transmitter release, but no interference with presynaptic autoinhibition of the release. Neither of these alterations seemed to account for the cognitive impairment assessed. PMID- 12759125 TI - Dimenhydrinate produces a conditioned place preference in rats. AB - Dimenhydrinate (DMH; trade names Gravol and Dramamine) is a compound of diphenhydramine (DP) and 8-chlorotheophylline in equimolar ratios. DMH has been reported to be abused by humans for its euphoric and hallucinogenic properties but few studies have evaluated its reinforcing effects in animals. To evaluate the hypothesis that DMH and its constituents DP and 8-chlorotheophylline are rewarding in animals, rats were tested for conditioned place preference (CPP). The paradigm consisted of pre-exposure (three 15-min sessions of access to both sides of the chamber), conditioning [eight 30-min pairings of one side with drug (four sessions) and, on alternate days, the other side with vehicle (four sessions)] and test phases (three 15-min sessions of access to both sides of the chamber). Significant preferences for the drug-paired location were found on test session one after conditioning with 60.0, but not 25.0, 40.0 or 50.0 mg/kg of DMH, and after conditioning with 37.8 but not 27.0 or 32.4 mg/kg of DP. No preference was found after conditioning with 23.0, 27.6 or 32.2 mg/kg of 8 chlorotheophylline. All three drugs stimulated locomotor activity during conditioning sessions and DMH and DP showed sensitization over conditioning sessions. DMH doses that showed sensitization (25.0 and 40.0 mg/kg) were lower than the dose (60.0 mg/kg) that produced a CPP revealing a dissociation of locomotor stimulating versus rewarding effects. Results reveal that DMH and DP have rewarding properties, although the molar equivalent dose-response curve for DP appeared to be further to the right than that for DMH. Future investigations into the neurotransmitter systems modulating this effect are awaited. PMID- 12759124 TI - Amphetamine-modified acoustic startle responding and prepulse inhibition in adult and adolescent alcohol-preferring and -nonpreferring rats. AB - Selective breeding has been used to develop the alcohol-preferring (P) and nonpreferring (NP) rats, with the P rat having lower CNS levels of dopamine (DA) and reduced DA innervation in the nucleus accumbens compared with the NP rat. The acoustic startle response (ASR) and prepulse inhibition (PPI) of the ASR are experimental behaviors altered by DA agonists. We examined whether functional differences in amphetamine (AMPH)-modified ASR and PPI exist between P and NP rats. AMPH [0.0 (saline), 1.0, 2.0, or 4.0 mg/kg] was injected 15 min prior to placement into a startle apparatus. After a 5-min habituation period, rats were given approximately twelve 95-, 105-, or 115-dB white-noise burst (ASR) and PPI trials. As adults, P rats were sensitive to AMPH potentiation of the ASR to a greater extent than NP rats. During adolescence, P and NP rats had similar levels of AMPH-potentiated ASR. As adults, NP rats displayed potentiated, rather than disrupted, PPI at the 1.0-mg/kg dose, whereas P rats displayed the expected disrupted PPI at the 4.0-mg/kg dose. As adolescents, NP rats did not display significant differences in PPI after AMPH, whereas P rats displayed dose dependent disruption of PPI, which was significant at the 4.0-mg/kg dose. The limited effect of AMPH on increasing the ASR and the presence of AMPH-potentiated PPI at the lowest dose in the adult NP rat suggests reduced functioning of the interactions between DA circuits and the neurocircuitry mediating the ASR and PPI, compared with P rats. However, the neurocircuitry mediating PPI does not appear to be fully developed in the adolescent NP rat. The present findings also indicate that lower levels of DA content and immunoreactive fibers in the P rat may not reflect reduced DA neuronal activity, because the P rat displayed AMPH potentiated ASR, and, at the highest dose, AMPH disruption of PPI during both adulthood and adolescence. PMID- 12759126 TI - Meal pattern changes associated with temporomandibular joint inflammation/pain in rats; analgesic effects. AB - Establishing a valid animal model to study temporomandibular joint (TMJ) pain has proven extremely difficult. Using complete Freund's adjuvant (CFA) to induce TMJ inflammation, we recently showed that meal pattern analysis could be used as a noninvasive biological marker to study TMJ pain in an animal model. The purpose of this study was to further validate our animal model by determining whether aspects of CFA-induced TMJ inflammation/pain are reversed with ibuprofen (IBU) treatment. In the first trial, 48 male rats were used and in the second trial, 32 female ovariectomized rats, given 17beta-estradiol replacement, were used. The rats were assigned to one of four groups: control (CON-CON); control+IBU (CON+IBU); CFA-CON; and CFA+IBU. In the male trial, CFA injection (P<.01) caused TMJ swelling and chromodacryorrhea (CFA-CON); IBU eliminated these changes in the CFA+IBU group. Meal pattern analysis showed the pertinent CFA-induced change and the IBU effect was that meal duration was increased in the CFA-CON group (P<.01), but normal in the CFA+IBU-treated group on the first, but not second, day postinjection. In the female trial, CFA increased TMJ swelling, but did not cause significant chromodacryorrhea (CFA-CON); IBU eliminated swelling in the CFA+IBU group. Meal duration was increased (P<.01) in the CFA-CON group, but was normal in the CFA+IBU-treated group on both the first and second days postinjection. In both trials, interleukin-1beta (IL-1beta) levels were increased similarly in CFA CON and CFA+IBU groups (P<.01). This study shows that CFA-induced TMJ inflammation/pain can cause changes in meal patterns (i.e., meal duration), which may be used as a behavioral marker for TMJ inflammation/pain. PMID- 12759127 TI - Heritable differences in the effects of amphetamine but not DOI on startle gating in albino and hooded outbred rat strains. AB - Sensorimotor gating, measured by prepulse inhibition (PPI) of the startle reflex, is reduced in schizophrenia patients and in rats treated with dopamine (DA) agonists. Strain and substrain differences in the sensitivity to the PPI disruptive effects of DA agonists may provide insight into the basis for human population differences in sensorimotor gating. We reported heritable differences in sensitivity to the PPI-disruptive effects of the D1/D2 agonist apomorphine (APO) in Harlan Sprague-Dawley (SDH) and Long-Evans (LEH) rats, offspring (F1) of an SDHxLEH cross, and subsequent offspring (N2) of an SDHxF1 cross. In this study, we assessed the neurochemical specificity of this heritable phenotype across parental SDH and LEH strains, and their F1 and N2 offspring, based on their sensitivity to the PPI-disruptive effects of the indirect DA agonist D amphetamine (AMPH) and the 5HT2A agonist DOI. AMPH sensitivity followed a gradient of SDH>N2>F1>LEH, consistent with past findings with APO. DOI sensitivity did not differ across strains or generations. These findings demonstrate that the heritable phenotype in this model is not specific to a particular compound (APO), and reflects physiological differences in the DAergic, but not serotonergic, regulation of PPI. PMID- 12759128 TI - Facilitation of electrical brain self-stimulation behavior by abused solvents. AB - Animal models are needed to study the abuse-related behavioral and pharmacological effects of inhaled solvents. Previous studies have suggested that intracranial self-stimulation techniques may be successfully adapted for testing the effects of solvent exposure. The present study aimed to assess the effects of toluene, cyclohexane, acetone, and petroleum benzine (a widely used mixture of hexanes and heptanes) in rats trained to lever press or nose-poke for electrical stimulation delivered through electrodes implanted into the medial forebrain bundle. It was found that toluene, cyclohexane, and benzine but not acetone, increased rates of responding, particularly at the lower stimulation intensities. In another set of experiments utilizing an auto-titration procedure, all tested solvents significantly reduced self-stimulation thresholds. However, only for toluene and benzine were these effects observed at the exposure levels that did not impair rates of operant performance. There may not be such a clear separation of effects for acetone and cyclohexane. Thus, toluene and benzine appear to selectively affect brain reward systems in a manner similar to that for most other abused drugs. Data from intracranial self-stimulation studies of solvents may be useful in abuse potential assessment of individual compounds and for examining neural and behavioral processes involved in inhalant abuse. PMID- 12759129 TI - Norepinephrine regulates locomotor hyperactivity in the mouse mutant coloboma. AB - An imbalance between dopaminergic and noradrenergic systems is implicated in hyperactivity disorders such as attention deficit hyperactivity disorder (ADHD) and Tourette syndrome. We have identified the mouse mutant coloboma as an animal model for examining the neurological basis of hyperactivity. Coloboma mice exhibit spontaneous locomotor hyperactivity that is a result of a reduction in SNAP-25, a presynaptic protein that regulates exocytotic release. These mice exhibit an imbalance in catecholamine regulation whereby brain dopamine (DA) utilization is reduced while norepinephrine (NE) concentrations are significantly increased. Further, calcium-dependent NE release was also increased in these hyperactive mice, despite the reduction in SNAP-25. To determine the role of NE in the expression of hyperactivity, brain NE concentrations were reduced using the specific noradrenergic neurotoxin DSP-4 [N-(2-chloroethyl)-N-ethyl-2 bromobenzylamine hydrochloride]. DSP-4 treatment specifically decreased NE concentrations, but had no effect on brain DA or serotonin. Depletion of NE by DSP-4 through either systemic or central administration significantly reduced the locomotor activity in coloboma mice. These results suggest that NE regulation in the CNS plays an important role in the expression of hyperactivity in this mouse model, consistent with results of human studies and current models of ADHD. PMID- 12759130 TI - Effects of imidacloprid metabolites on habituation in honeybees suggest the existence of two subtypes of nicotinic receptors differentially expressed during adult development. AB - Habituation of the proboscis extension reflex (PER) in honeybees (Apis mellifera) is age-dependent. Very young bees (< or =7 days old) require significantly less trials to abolish the response to multiple sucrose stimulations than older bees (> or =8 days old). A nicotinic agonist, imidacloprid, modifies this behaviour by increasing the number of trials in < or =7-day-old bees and by decreasing it in older bees [Neurobiol. Learn. Mem. 76 (2001) 183.]. Here we tested our hypothesis that this effect is associated with a differential expression of two subtypes of nicotinic acetylcholine receptors (nAChRs). By testing the effects of six metabolites of imidacloprid, we show that two of them, olefin and 5-hydroxy imidacloprid, modify the number of trials needed to habituate the PER in a contrasting manner. Olefin increases the number of trials in both age groups, whereas 5-hydroxy-imidacloprid decreases the number of trials, but only in 8-day old individuals. We conclude that olefin and 5-hydroxy-imidacloprid are specific agonists of two subtypes of an nAChR that are differentially expressed during adult maturation of young honeybees. Olefin is the agonist of an nAChR expressed in both age groups, whereas 5-hydroxy-imidacloprid is the agonist of a late-onset nAChR that is activated in 8-day-old bees. The implications of this finding for the honeybee biology are discussed. PMID- 12759131 TI - Flavor preferences conditioned by intragastric ethanol with limited access training. AB - In a prior study, ad libitum fed rats learned a strong preference (90%) for a flavored saccharin solution (conditioned stimulus, CS+) paired with concurrent intragastric (IG) infusions of 5% ethanol over another flavor (CS-) paired with water infusions in unlimited access sessions (22 h/day). The present study expanded the investigation of ethanol-conditioned preferences to limited access sessions (30 min/day). Experiment 1 revealed that ad lib or food-restricted rats failed to develop a CS+ preference using the same CS solutions (0.05% Kool Aid+0.2% saccharin) and IG infusions that were effective with long-term training. Experiments 2 and 3 mimicked the parameters from a report of successful ethanol conditioning in deprived rats: ethanol (0.5 g/kg) or water was infused intragastrically 5 min before access to sweetened CS solutions flavored with HCl or NaCl. Rats learned to prefer the ethanol-paired CS+ when the flavors were mixed with 5% sucrose but not when mixed with 0.2% saccharin. Experiment 4 revealed that 5% sucrose solutions flavored with 0.25% Kool Aid also supported flavor preference conditioning by IG ethanol (0.5 g/kg). CS+ preferences were obtained in rats trained with ethanol infused 5 min before or concurrent with CS+ intake, but not in rats trained with ethanol infused 30 min before CS+ intake. These data confirm that flavor preferences can be conditioned by IG ethanol using a limited access procedure. However, in contrast to 22 h/day training, 30 min/day training requires more intense CS flavors and a nutritive sweetener. The preference reinforcing actions of ethanol may develop slowly and are thus most effective with long training sessions or when intense CS flavors are used in short training sessions. PMID- 12759132 TI - Agonist/antagonist properties of nalbuphine, butorphanol and (-)-pentazocine in male vs. female rats. AB - To determine whether sex differences in the effects of mixed-action opioids could be due to differential activity at mu or kappa receptors, agonist/antagonist properties of nalbuphine, butorphanol and (-)-pentazocine were compared in male vs. female rats using a diuresis test. In water-loaded rats (2-h test), nalbuphine and (-)-pentazocine dose-dependently increased urination similarly in both sexes, whereas butorphanol increased urination more in females than in males on a ml/kg basis. The diuretic effects of all three opioids were at least partially blocked by the kappa receptor-selective antagonist nor-binaltorphimine (nor-BNI, 5 mg/kg) in both sexes. Kappa receptor-mediated antagonism of diuresis induced by U69,593 (0.56 mg/kg) was only observed with butorphanol in males. In water-loaded rats (1-h test), nalbuphine did not suppress, and butorphanol and ( )-pentazocine significantly suppressed urination in males only; all three mixed action opioids dose-dependently blocked the antidiuretic effect of the selective mu agonist fentanyl (0.056 mg/kg) in both sexes. The ability of nalbuphine and ( )-pentazocine to block fentanyl-induced antidiuresis was not affected by pretreatment with nor-BNI in either sex. In contrast, the ability of butorphanol to block fentanyl-induced antidiuresis was attenuated by pretreatment with nor BNI in males but not in females. These results suggest that sex differences in the effects of these mixed-action opioids are primarily due to their greater relative efficacy at the mu receptor in male than in female rats; butorphanol also may have greater efficacy at kappa receptors in females than in males. PMID- 12759133 TI - Prevention by dietary (n-6) polyunsaturated phosphatidylcholines of intrahepatic cholestasis induced by cyclosporine A in animals. AB - Previous findings showed that dietary (n-6) polyunsaturated phosphatidylcholines (vegetable lecithin) could efficiently prevent intrahepatic cholestasis induced by cyclosporine A in rats. Mechanistic studies showed that expressions in rat liver of Na(+), K(+)-ATPase, Ca(2+), Mg(2+)-ATPase and F-actin were both decreased by drug administration and both enhanced by (n-6) lecithin enriched diet. There is a possible direct effect of phosphatidylcholines, vectors of polyunsaturated fatty acids provided by the metabolism of the dietary lecithin, on the aforesaid hepatic parameters. Such modulations by drug and diet result in reversed modifications of membrane composition and fluidity. Final outcome is decreased and enhanced bile lipid secretion by cyclosporine and vegetable lecithin enriched diet respectively. Moreover, we advance the hypothesis of a bypass process including a separate and functional actin-independent way for the non micellar and phospholipid-dependent secretion of bile lipids. The relationships between the ATPases, the microfilament components such as F-actin and the different transporters still remain to be clarified. Furthermore, one can speculate on beneficial effects in humans of diets enriched in vegetable lecithins that might prevent cholestasis induced by cyclosporine A. PMID- 12759134 TI - Prolonged treatment with prostaglandin E1 increases the rate of lipolysis in rat adipocytes. AB - Prolonged treatment of adipocytes with certain inhibitors of lipolysis, including N(6)-phenylisopropyl adenosine (PIA) and prostaglandin E(1) (PGE(1)) leads to down-regulation of G(i). Prolonged treatment with PIA increases the rate of lipolysis, and we have reported that tumor necrosis factor-alpha (TNF alpha) stimulates lipolysis by down-regulating G(i). To determine the relationship between G(i) concentration and lipolysis, we have investigated the effect of two other acute inhibitors of lipolysis; PGE(1), which down-regulates G(i), and nicotinic acid (NA), which does not down-regulate G(i). Rat adipocytes were incubated with PIA (300 nM), PGE(1) (3 microM) or nicotinic acid (1 mM) for 24 h. The rate of lipolysis (glycerol release) was increased approximately 2 to 3-fold in PIA-treated cells, and in PGE(1)-treated cells. Conversely, the rate of lipolysis was not altered by the prolonged nicotinic acid treatment. These findings support the hypothesis that the rate of lipolysis in adipocytes is determined, at least partly, by the cellular concentration of G(i) proteins. PMID- 12759135 TI - Endomorphin-2, deltorphin II and their analogs suppress formalin-induced nociception and c-Fos expression in the rat spinal cord. AB - In this study, we evaluated the effects of intrathecally administered agonists of mu- and delta-opioid receptor and their analogs on the pain-induced behavior and expression of c-Fos immunoreactivity in the spinal cord, elicited by intraplantar injection of 12% formalin to the hindpaw of the rat. Previous report from our laboratory and other author's study indicated that intrathecal administration of mu agonists morphine and endomorphin-2 and delta-opioid agonist deltorphin II produced a dose-dependent antinociceptive effects in acute and inflammatory pain. In this study, intrathecal injection of morphine (10 microg), endomorphin-2 (5 microg) and its analog Dmt-endomorphin-2 (10 microg) significantly decreased the formalin-induced pain behavior, and lowered a number of c-Fos positive neurons in the laminae I, II and III of the spinal cord by about 40%, 30% and 40%, respectively. Significant reduction of formalin-induced behavioral responses was also observed after i.th. administration of deltorphin II (15 microg) and its analog ile-deltorphin II (15 microg). Agonists of delta-opioid receptor significantly reduced a number of c-Fos positive neurons by about 28% and 40%, respectively. Analog of endomorphin-2 and analog of deltorphin II suppressed more potently expression of c-Fos in the dorsal horn of the spinal cord than the parent peptides. Our study indicates that new analogs of mu- and delta-opioid receptor exhibit strong antinociceptive potency similar or even higher than the parent peptides, and that their effect is positively correlated with the inhibition of c-Fos expression. PMID- 12759136 TI - Agmatine attenuates neuropathic pain in rats: possible mediation of nitric oxide and noradrenergic activity in the brainstem and cerebellum. AB - Effect of agmatine (10-400 mg/kg) on neuropathic pain in a rat model produced by loose ligatures around the common sciatic nerve was studied. The involvement of possible alterations in nitric oxide (NO) levels [measured as its stable metabolites nitrate + nitrite] and in noradrenergic activity [measured as norepinephrine and 3-methoxy-4-hydroxyphenylethylene glycol (MHPG) levels] in this effect was also investigated biochemically in the brainstem and cerebellum. Agmatine increased the neuropathic pain threshold at 300 and 400 mg/kg. There was almost a twofold increase in nitrate + nitrite levels in the brainstem and cerebellum of the rats with neuropathic pain and agmatine decreased the high nitrate + nitrite levels only in the brainstem at 300 mg/kg and both in the brainstem and cerebellum at 400 mg/kg. Ligation of sciatic nerve resulted in almost twofold increase in norepinephrine and MHPG levels only in the brainstem of the rats. Agmatine decreased MHPG levels at 300 and 400 mg/kg, however it decreased norepinephrine levels only at the higher dose. These findings indicate that agmatine decreases neuropathic pain, an effect which may involve the reduction of NO levels and noradrenergic activity in the brain. PMID- 12759137 TI - Effects of lomefloxacin and norfloxacin on pancreatic beta-cell ATP-sensitive K(+) channels. AB - In patients administered lomefloxacin alterations in blood glucose concentrations have been observed in some cases and lomefloxacin has previously been shown to augment insulin release from rat pancreatic islets at micromolar concentrations. The aim of the present study was to compare the effects of two structurally related fluoroquinolones, lomefloxacin and norfloxacin, on ATP-sensitive K(+) (K(ATP)) currents from the clonal insulinoma cell line RINm5F using the whole cell configuration of the patch-clamp technique. The application of lomefloxacin concentration-dependently blocked K(ATP) currents from RINm5F cells with a half maximally inhibitory concentration of 81 microM, whereas the application of norfloxacin (at concentrations up to 300 microM) had only minor effects on K(ATP) currents. Block of pancreatic beta-cell K(ATP) currents could be mediated by interaction of lomefloxacin either with the regulatory subunit (SUR1) or with the pore-forming subunit (Kir6.2). We favour the latter hypothesis, since some fluoroquinolones have recently been shown to block the pore-forming subunit of the cardiac rapid delayed rectifier K(+) current I(Kr) (which is encoded by HERG (human ether-a-go-go-related gene)). Thus, as demonstrated for cardiac HERG channels in previous studies and for pancreatic beta-cell K(ATP) channels in the present study, fluoroquinolones differ markedly in their potencies to inhibit K(+) channel activity. PMID- 12759139 TI - Leukotactin-1-induced ERK activation is mediated via Gi/Go protein/PLC/PKC delta/Ras cascades in HOS cells. AB - Recently cloned leukotactin-1 (Lkn-1) that belongs to CC chemokine family has not been characterized. To understand the intracellular events following Lkn-1 binding to CCR1, we investigated the activities of signaling molecules in response to Lkn-1 in human osteogenic sarcoma cells expressing CCR1. Lkn-1 stimulated cells showed elevated phosphorylation of extracellular signal-related kinases (ERK1/2) with a distinct time course. ERK activation was peaked in 30 min and 12 h showing biphasic activation of ERK. Pertussis toxin, an inhibitor of G(i)/G(o) protein, and phospholipase C (PLC) inhibitor blocked Lkn-1-induced activation of ERK. Protein kinase C delta (PKC delta) specific inhibitor rottlerin inhibited ERK activation in Lkn-1-stimulated cells. The activities of PLC and PKC delta were also enhanced by Lkn-1 stimulation. Dominant negative Ras inhibited activation of ERK. Immediate early response genes such as c-fos and c myc were induced by Lkn-1 stimulation. Lkn-1 affected the cell cycle progression by cyclin D(3) induction. These results suggest that Lkn-1 activates the ERK pathway by transducing the signal through G(i)/G(o) protein, PLC, PKC delta and Ras, and it may play a role for cell proliferation, differentiation, and regulation of gene expression for other cellular processes. PMID- 12759138 TI - Non-steroidal anti-inflammatory drugs affect the methotrexate transport in IEC-6 cells. AB - Methotrexate (MTX) is used not only for the cancer chemotherapy but also for the treatment of rheumatic disease, often together with non-steroidal anti inflammatory drugs (NSAIDs). MTX is actively cotransported with H(+) in the small intestine, mediated by a reduced folate carrier (RFC). The coadministration of some NSAIDs with MTX to rats caused a decrease of MTX absorption through the small intestine. This may be due to the uncoupling effect of oxidative phosphorylation of the NSAIDs. The present study investigated whether flufenamic acid, diclofenac and indomethacin, NSAIDs, decreased ATP content of rat-derived intestinal epithelial cell line IEC-6 cells and affected the MTX transport in IEC 6 cells. The MTX uptake in IEC-6 cells was dependent on medium pH and maximum around pH 4.5-5.5. The MTX uptake was composed of a transport inhibited by 4, 4' diisothiocyanostilbene-2, 2'-disulfonic acid (DIDS) and a non-saturable one. The DIDS-sensitive component in the MTX uptake showed a saturation kinetics (Michaelis-Menten constant (Km): 3.91 +/- 0.52 microM, Maximum velocity (Vmax): 94.66 +/- 6.56 pmol/mg protein/5 min). The cellular ATP content in IEC-6 cells decreased significantly at 30 min after the cells were started to incubate with the NSAIDs (250 microM flufenamic acid, 500 microM diclofenac and 500 microM indomethacin). The MTX uptake in IEC-6 cells in the presence of the NSAIDs decreased with the reduction of cellular ATP content and showed a good correlation with the ATP content (correlation coefficient: 0.982). Thus it seems likely that the ATP content in IEC-6 cells with the NSAIDs decreased due to the uncoupling effect of oxidative phosphorylation of the NSAIDs, resulting in the inhibition of the secondary active transport of MTX in IEC-6 cells. The present results also suggest that IEC-6 cells are useful to evaluate the drug interaction relating to this carrier system. PMID- 12759140 TI - Hyperforin inhibits vesicular uptake of monoamines by dissipating pH gradient across synaptic vesicle membrane. AB - Extracts of Hypericum perforatum (St. John's wort) have antidepressant properties in depressed patients and exert antidepressant-like action in laboratory animals. The phloroglucinol derivative hyperforin has become a topic of interest, as this Hypericum component is a potent inhibitor of monoamines reuptake. The molecular mechanism by which hyperforin inhibits monoamines uptake is yet unclear. In the present study we try to clarify the mechanism by which hyperforin inhibits the synaptic vesicle transport of monoamines. The pH gradient across the synaptic vesicle membrane, induced by vacuolar type H(+)-ATPase, is the major driving force for vesicular monoamines uptake and storage. We suggest that hyperforin, like the protonophore FCCP, dissipates an existing Delta pH generated by an efflux of inwardly pumped protons. Proton transport was measured by acridine orange fluorescence quenching. Adding Mg-ATP to a medium containing 130 mM KCl and synaptic vesicles caused an immediate decrease in fluorescence of acridine orange and the addition of 1 microM FCCP abolished this effect. H(+)-ATPase dependent proton pumping was inhibited by hyperforin in a dose dependent manner (IC(50) = 1.9 x 10(-7) M). Hyperforin acted similarly to the protonophore FCCP, abolishing the ATP induced fluorescence quenching (IC(50) = 4.3 x 10(-7) M). Hyperforin and FCCP had similar potencies for inhibiting rat brain synaptosomal uptake of [3H]monoamines as well as vesicular monoamine uptake. The efflux of [3H]5HT from synaptic vesicles was sensitive to both drugs, thus 50% of preloaded [3H]5HT was released in the presence of 2.1 x 10(-7) M FCCP and 4 x 10(-7) M hyperforin. The effect of hyperforin on the pH gradient in synaptic vesicle membrane may explain its inhibitory effect on monoamines uptake, but could only partially explain its antidepressant properties. PMID- 12759141 TI - Antinociceptive mechanisms of orally administered decursinol in the mouse. AB - Antinociceptive profiles of decursinol were examined in ICR mice. Decursinol administered orally (from 5 to 200 mg/kg) showed an antinociceptive effect in a dose-dependent manner as measured by the tail-flick and hot-plate tests. In addition, decursinol attenuated dose-dependently the writhing numbers in the acetic acid-induced writhing test. Moreover, the cumulative response time of nociceptive behaviors induced by an intraplantar formalin injection was reduced by decursinol treatment during the both 1st and 2nd phases in a dose-dependent manner. Furthermore, the cumulative nociceptive response time for intrathecal (i.t.) injection of TNF-alpha (100 pg), IL-1 beta (100 pg), IFN-gamma (100 pg), substance P (0.7 microg) or glutamate (20 microg) was dose-dependently diminished by decursinol. Intraperitoneal (i.p.) pretreatment with yohimbine, methysergide, cyproheptadine, ranitidine, or 3,7-dimethyl-1-propargylxanthine (DMPX) attenuated inhibition of the tail-flick response induced by decursinol. However, naloxone, thioperamide, or 1,3-dipropyl-8-(2-amino-4-chloro-phenyl)-xanthine (PACPX) did not affect inhibition of the tail-flick response induced by decursinol. Our results suggests that decursinol shows an antinociceptive property in various pain models. Furthermore, antinociception of decursinol may be mediated by noradrenergic, serotonergic, adenosine A(2), histamine H(1) and H(2) receptors. PMID- 12759143 TI - Increase in complement component C3 is an early response to experimental magnesium deficiency in rats. AB - The importance of the inflammatory process in the pathology of experimental Mg deficiency has been reconsidered but the sequence of events leading to inflammatory response remains unclear. In this study, the effect of Mg-deficiency on complement system by measuring total C3 concentration, mRNA abundance for rat pre-pro complement C3 in liver by RT-PCR, complement haemolytic activity and C3 activation by Western Blot was studied. Weaning male Wistar rats were fed either Mg-deficient or control experimental diets for 2 or 8 days. At 8 days, a characteristic inflammatory response of Mg-deficiency including hyperaemia, leukocytosis and enlarged spleen was accompanied by an increase in the total C3 quantity in plasma. Moreover, at 8 days, RT-PCR analysis indicated higher level of mRNA rat pre-pro complement C3 in liver from Mg-deficient rats compared to control rats. Even if the inflammatory syndrome was not observed in rats after 2 days, total plasma C3 was shown to be significantly increased as compared to total plasma C3 level in control rats. Because of the high variability of complement haemolytic activity values in Wistar rats, weaning male Sprague-Dawley rats were used in a second experiment. At 8 days, the inflammatory response of Sprague-Dawley rats was accompanied by an increase in total C3 quantity and by a higher haemolytic activity. The Western Blot technique failed to display distinct bands resulting from C3 cleavage in plasma from Mg-deficient rats. Since, the complement C3 is a positive acute phase reactant, the elevation of C3 indicates that the modification of inflammatory response is an early event of Mg deficiency. However, complement activation does not appear to be involved in the acute phase of the deficiency. PMID- 12759142 TI - Arachidonyl dopamine as a ligand for the vanilloid receptor VR1 of the rat. AB - The vanilloid receptor VR1 is a nonspecific Ca(2+) channel, expressed in sensory neurons in the peripheral nervous system and in various brain regions, which is believed to be an important molecular integrator of several chemical (acid, vanilloids) and physical stimuli (heat) that cause pain. Recently, several endogenous ligands for VR1 have been identified such as arachidonyl ethanolamide (anandamide) and the more potent arachidonyl dopamine (AA-DO). Here, we further characterize AA-DO as a ligand for rat VR1, heterologously expressed in CHO and HEK293 cells. AA-DO inhibited the binding of [3H]RTX to VR1 with a K(d) value of 5.49 +/- 0.68 microM and with positive cooperativity (p = 1.89 +/- 0.27), indicating that AA-DO was about 5-fold more potent than anandamide in this system. The K(d) (9.7 +/- 3.3 microM), and p values (1.54 +/- 0.04) for the binding of AA-DO to spinal cord membranes are in good correlation with the CHO VR1 data. AA-DO stimulated 45Ca(2+) uptake on CHO-VR1 and HEK-VR1 cells with EC(50) values of 4.76 +/- 1.43 and 7.17 +/- 1.64 microM and Hill coefficients of 1.28 +/- 0.11 and 1.13 +/- 0.13, respectively, consistent with the binding measurements. In contrast to anandamide, AA-DO induced virtually the same level of 45Ca(2+) uptake as did capsaicin (90 +/- 6.6% in the CHO cells expressing VR1 and 89.3 +/- 9.4% in HEK293 cells expressing VR1). In a time dependent fashion following activation, AA-DO partially desensitized VR1 both in 45Ca(2+) uptake assays (IC(50) = 3.24 +/- 0.84 microM, inhibition is 68.5 +/- 6.85%) as well as in Ca(2+) imaging experiments (35.8 +/- 5.1% inhibition) using the CHO-VR1 system. The extent of desensitization was similar to that caused by capsaicin itself. We conclude that AA-DO is a full agonist for VR1 with a potency in the low micromolar range and is able to significantly desensitize the cells in a time and dose dependent manner. PMID- 12759144 TI - Activins, inhibins, and follistatins in reproduction. PMID- 12759145 TI - Expression and function of placenta growth factor: implications for abnormal placentation. AB - OBJECTIVE: Essential requirements for successful gestation include the coordinated growth and differentiation of the placenta and the development of a functional placental vasculature. However, relatively little is known about factors that are responsible for regulating these functions. One angiogenic growth factor that might be involved in regulating both vascular endothelial cell and trophoblast function is placental growth factor (PGF). METHODS: Current published reports were surveyed and our own work was reviewed to highlight the expression, function, and potential significance of PGF at the human maternal fetal interface. RESULTS: PGF is highly expressed in trophoblasts during normal pregnancy, and its expression is significantly decreased in preeclampsia, an obstetric complication presumed to be associated with placental bed hypoxia and ischemia. In agreement with this, in vitro trophoblast expression of PGF can be down-regulated by low oxygen tension. The cognate receptor for PGF, fms-like tyrosine kinase receptor, is expressed on trophoblasts as well as vascular endothelial cells, suggesting that it has autocrine and paracrine functions. Accordingly, PGF can regulate proliferation in first trimester trophoblasts, apoptosis in term trophoblasts, and it can directly or indirectly regulate vascular growth, maturation, and permeability. CONCLUSIONS: Many obstetric complications, most notably preeclampsia, are associated with aberrant trophoblast function and inadequate or dysfunctional vasculature within the developing placenta. The ability of PGF to influence trophoblast and vascular endothelial cells provides clear impetus for further studies to investigate the biological and clinical significance of PGF in normal and abnormal human pregnancies. PMID- 12759146 TI - Effects of long-term high-altitude hypoxia on myocardial protein kinase A activity and troponin I isoforms in fetal and nonpregnant sheep. AB - OBJECTIVE: In fetal sheep, we found that the augmentation of cardiac contractility by beta-adrenergic receptor (beta-AR) stimulation was reduced after exposure to long-term hypoxia. However, cyclic adenosine monophosphate (cAMP) production after beta-AR stimulation was higher in long-term hypoxic fetal sheep than in normoxic ones. Therefore, we studied the potential role of changes in myocardial protein kinase A (PKA) activity and troponin I (TnI) isoforms in fetal and nonpregnant sheep exposed to approximately 112 days of hypoxia at high altitude (3820 m). METHODS: Resting and maximally stimulated (by cAMP) PKA activity was measured by phosphorylation of the artificial peptide, Kemptide. Specificity was confirmed by inhibition with PKI, a specific PKA inhibitor. For TnI isoforms, sodium dodecyl sulfate-polyacrylamide gel electrophoresis was used to resolve the proteins. We used monoclonal anti-cardiac TnI antibody (clone C5), which also cross-reacted with slow skeletal muscle TnI, to identify TnI isoforms. RESULTS: For the fetal hearts, resting PKA activity was significantly higher in the high-altitude group than the control group, but total PKA activity was not different between the normoxic and hypoxic groups. In the adult hearts, no significant difference was observed in either resting or total PKA activity between normoxic and hypoxic groups. For both the fetal and adult sheep, the predominant TnI was the cardiac isoform, and hypoxic exposure produced no change in the TnI isoform composition. CONCLUSIONS: Neither a reduction in PKA activity nor a change in TnI isoforms could explain the reduction in beta-receptor augmentation of cardiac contractility in fetal sheep exposed to long-term hypoxia. PMID- 12759147 TI - A new ultrasound technique to measure the isovolumetric contraction time as an index of cardiac contractility: fetal lamb validation. AB - OBJECTIVE: We developed a noninvasive Doppler technique for measuring fetal cardiac isovolumetric contraction time (ICT). The purpose of this study was to determine how well our method reflects real cardiac performance using fetal lamb as an instrumented model. METHODS: The true ICT was measured by simultaneous recording of the pressure waves of the left ventricle and ascending aorta. The maximum first derivative of the left ventricular pressure wave (Max dp/dt) was calculated. The Doppler ICT was measured in the appropriately filtered Doppler cardiac signals. Positive and negative inotropic agents were administered to change the cardiac contractility. RESULTS: There was an inverse relationship between the Doppler ICT and the Max dp/dt. Excellent linear correlation was found an absolute value and changes from control value between the true ICT and the Doppler ICT (r = 0.959, r = 0.962). CONCLUSIONS: The Doppler ICT measurement provides useful information about changes in ventricular performance. PMID- 12759148 TI - Continuous systolic blood pressure monitoring by the difference in electrocardiogram and pulse oximetry in near-term, exteriorized goat fetuses. AB - OBJECTIVE: Our purpose was to determine whether systolic arterial pressure, as predicted from the delay time from the electrocardiogram to pulse oximeter waveform, correlates with directly measured systolic blood pressure. METHODS: Six Japanese Saanen goats at 120-125 days' gestation were used. Experiments were conducted in acutely operated animals under anesthesia. We placed catheters in the carotid artery and jugular vein, electrodes on the chest, and a pulse oximeter probe on the tongue. Systolic blood pressure (Psm), fetal heart rate (FHR), delay time (Td), and pulse oximeter waveform were monitored continuously, and the data were stored. Systolic blood pressure was estimated (Pse) by the simple linear equation of Td. Fetal blood pressure was decreased 10% by hydralazine and blood withdrawal. Correlation coefficients and the agreement between Psm and Pse were determined. RESULTS: A significant correlation (P <.01) with good bias values (< 8 mmHg) and precision values (< 5 mmHg) was obtained between Psm and Pse in all animals. There was also a significant correlation between Psm and Pse with high coefficients for hydralazine (r(2) = 0.82 +/- 0.12) and for withdrawal (r(2) = 0.85 +/- 0.15). The correlations became slightly scattered in acidemia (pH < 7.25). CONCLUSIONS: This technique has the potential to provide information on fetal blood pressure during labor and delivery on a beat-by-beat basis, which will aid in the interpretation of electronic FHR recordings. PMID- 12759149 TI - Estrogen selectively up-regulates eNOS and nNOS in reproductive arteries by transcriptional mechanisms. AB - OBJECTIVE: To determine the mechanism(s) whereby daily and acute estradiol-17beta (E(2)beta) exposure modifies endothelium-derived nitric oxide synthase (eNOS) and vascular smooth muscle (VSM) neuronal nitric oxide synthase (nNOS) in reproductive and nonreproductive arteries and to localize NOS isoform expression within the vessel wall. METHODS: Oophorectomized nonpregnant ewes received E(2)beta (1 microg/kg per day) or no E(2)beta for 5-6 days or acute E(2)beta (1 microg/kg) on day 6-7 with or without daily E(2)beta. Uterine, mammary, mesenteric, and femoral arteries were collected at completion of each study, adventitia were removed, and samples were frozen and stored at -80C. After separating endothelium and VSM, NOS isoform mRNA was measured using reverse transcription-polymerase chain reaction. VSM nNOS protein was determined by Western analysis. RESULTS: Basal eNOS and nNOS mRNA was greatest (P <.02) in reproductive artery endothelium and VSM, respectively. Daily E(2)beta was required for maximum uterine vascular responses to acute E(2)beta and was associated with increased reproductive artery endothelial eNOS mRNA (>1.5-fold, P <.02) and uterine VSM nNOS mRNA (>2.5-fold, P <.003) and protein (21%, P <.05). Acute E(2)beta in the presence and absence of daily E(2)beta also increased uterine eNOS 68% and 28% (P =.01), respectively, within 90 minutes but did not affect VSM nNOS. Mammary eNOS increased 71% only after E(2)beta withdrawal; VSM nNOS was unchanged. Neither NOS isoform was altered in nonreproductive arteries by daily or acute E(2)beta. CONCLUSIONS: Basal eNOS and nNOS isoform expression is greatest in arteries from reproductive tissues, and isoform responses to E(2)beta are cell specific and transcriptionally regulated. Furthermore, optimal uterine vascular responses to acute E(2)beta exposure require daily E(2)beta exposure that enhances basal NOS expression and abundance. PMID- 12759150 TI - Is low-dose streptozotocin in rats an adequate model for gestational diabetes mellitus? AB - OBJECTIVE: To examine the use of streptozotocin (SZ) in rats as a model for gestational diabetes mellitus (GDM). METHODS: We studied various doses of SZ, either as a single administration (30, 35, 40, or 50 mg/kg, intraperitoneally) on day 1 of pregnancy or as 2 low doses (30 and 20 or 30 and 30 mg/kg) administered 2 days before mating and on day 1 of pregnancy. We examined the effect on maternal and fetal glucose and insulin concentrations and on fetal weight on day 20 of pregnancy. In a second series of experiments, we studied two groups (SZ 30/20 and SZ 35) with fetal hyperinsulinemia on day 20 of pregnancy. We performed an intravenous glucose tolerance test (IVGTT) on day 20, and in separate groups we reassessed fetal weight and insulin concentrations at term (day 22). RESULTS: There was considerable variability in glucose concentrations with most SZ doses. Lower doses of SZ (30, 30/20, and 35 mg/kg) did not significantly increase maternal and fetal glucose levels, in contrast to higher doses of SZ (30/30 and 50 mg/kg). Fetuses were smaller on day 20 with all doses except SZ 30 and SZ 30/20; fetal insulin concentrations were elevated with SZ 30, 30/20, and 35. The IVGTT showed glucose intolerance in SZ 35 and SZ 30/20, but the insulin response was unaffected in either group. Fetuses were smaller on day 22 in both these SZ groups, whereas fetal insulin levels at term were not different compared with controls. CONCLUSIONS: Low-dose SZ is not a good model for GDM because of the high variability in glucose levels, the normal insulin response to a glucose load, the absence of fetal macrosomia, and the inconsistent effect on fetal insulin concentrations. PMID- 12759151 TI - The control of prostaglandin endoperoxide H-Synthase-2 expression in the human chorion laeve at term. AB - OBJECTIVE: Prostaglandin endoperoxide H synthase-2 (PGHS-2), the key enzyme of prostaglandin biosynthesis in gestational tissues, is expressed in the chorion laeve at term. We have determined the mechanisms that control the level of PGHS-2 mRNA in the chorion membrane in order to assess the significance of chorion derived prostaglandins in term labor. METHODS: Chorion membranes were collected after elective cesarean delivery (CD, n = 21) and after spontaneous labor (SL, n = 20) at term. The PGHS-2 gene transcription rate was measured by transcriptional run-on, and PGHS-2 mRNA and heterogeneous RNA (hnRNA) abundance was determined by quantitative real-time reverse transcriptase polymerase chain reaction. PGHS-2 mRNA stability, PGHS-2 hnRNA processing rate, and the short-term dynamics of the two RNA species were characterized in 0-24-hour-long tissue incubations. RESULTS: The transcriptional activity of the PGHS-2 gene predicted (P <.02) the abundance of PGHS-2 mRNA and hnRNA in individual tissues. PGHS-2 gene activity and hnRNA processing rate were not different in the CD and SL groups. PGHS-2 mRNA was constitutively stable before and after labor, and its abundance spontaneously increased sixfold in tissues incubated for 24 hours. At the same time, PGHS-2 gene activity decreased by 80% within 2 hours and rebounded to 60% of its initial level by 24 hours. CONCLUSIONS: PGHS-2 mRNA is highly stable, and its abundance is transcriptionally controlled in the chorion laeve at term. Labor is not associated with changing PGHS-2 gene activity. Endogenous factors drive PGHS-2 gene transcription in the chorion, and the stable PGHS-2 mRNA accumulates in the tissue at term. This accumulation has little or no impact on the timing of labor. PMID- 12759152 TI - Concentrations of serum total activin A and inhibin A in preterm and term labor patients: a cross-sectional study. AB - OBJECTIVE: To compare maternal serum levels of total activin A and inhibin A in preterm and term patients who are in labor or not in labor. METHODS: A cross sectional study compared activin A and inhibin A in the following groups of patients: preterm and in labor (n = 65), preterm and not in labor (n = 96), term and in labor (n = 65), and term and not in labor (n = 65). Preterm was defined as 23-34 weeks' gestation and term as 37-42 weeks' gestation. Labor was defined as regular contractions with progressive cervical change or an initial examination revealing cervical dilation of 1-3 cm with 50% effacement or more. Follistatin levels were analyzed in a subset of 12 patients from each group. Analytes were measured by two-site enzyme-linked immunosorbent assays. RESULTS: Activin A levels were higher in the preterm labor group (median 1.38 multiples of the median [MoM], interquartile range [IQR] 1.01 MoM) compared with the preterm nonlabor group (median 1.0 MoM, IQR 0.78 MoM, P <.05) and in the term labor group (median 1.37 MoM, IQR 1.74 MoM) compared with the term nonlabor group (median 1.0 MoM, IQR 0.87 MoM, P <.05). Inhibin A levels were higher in the preterm labor group (median 1.27 MoM, IQR 0.73 MoM) compared with the preterm nonlabor group (median 1.0 MoM, IQR 0.58 MoM, P <.05). Post-hoc analysis of activin A and inhibin A elevations in the preterm labor group revealed a significant effect only during 31-34 weeks' gestation. The total activin A:follistatin ratio, an indirect measure of free activin A, was similar between labor and nonlabor gestational age-matched patient groups. CONCLUSIONS: Levels of total activin A and inhibin A were increased in patients during labor; however, based on the moderate degree and narrow gestational age range of the increased levels, these analytes are not likely to be clinically useful in predicting preterm labor. PMID- 12759153 TI - Endometrial expression and secretion of activin A, but not follistatin, increase in the secretory phase of the menstrual cycle. AB - OBJECTIVE: Activin A is a growth factor expressed by human endometrium, and its biologic effects are counteracted by follistatin. We evaluate whether activin A and follistatin mRNA and peptide expression as well as protein secretion from human endometrium change throughout the menstrual cycle. METHODS: In 25 healthy fertile patients, uterine washing fluid was retrieved by hydrosonography. In a subgroup (n = 13), endometrial tissue samples were collected by hysteroscopy during the proliferative (n = 6) or secretory (n = 7) phase of the menstrual cycle. Activin and follistatin mRNA and peptide expression were evaluated by reverse transcriptase-polymerase chain reaction (RT-PCR) and by immunohistochemistry (IHC), respectively. Activin A and follistatin levels were assayed in uterine washing fluids by specific enzyme-linked immunosorbent assays and evaluated according to the endometrial thickness and menstrual cycle days. RESULTS: Both activin A and follistatin mRNAs were expressed by human endometrium, and their peptides immunolocalized both in proliferative and secretory endometrial epithelial and stromal cells. A significant increase in immunoreactive activin betaA but not in follistatin was observed in glandular epithelium during the secretory phase. Activin A but not follistatin was significantly (P <.0001) higher in the washing fluids collected during the secretory than proliferative phase of the menstrual cycle. In addition, a significant correlation was found between activin A, but not follistatin, and menstrual cycle days (P <.0001) or endometrial thickness (P <.0001). CONCLUSIONS: Both activin A and follistatin mRNAs are expressed by human endometrium; however, activin A but not follistatin peptide expression and secretion were increased in the secretory phase of the menstrual cycle, suggesting an important role in human endometrium. PMID- 12759154 TI - Generation of tumors in transgenic mice expressing the SV40 T antigen under the control of ovarian-specific promoter 1. AB - OBJECTIVE: The ovarian-specific promoter, OSP-1, which was cloned from the transcript of a rat retrovirus-like element specifically expressed in ovarian tissue, was tested for its ability to drive ovary-specific transcription in transgenic mice. METHODS: Transgenic mice were generated with the lacZ reporter gene (OSP-lacZ) or the early region of SV40 virus (OSP-TAg) placed under the control of the OSP-1 promoter. OSP-lacZ and OSP-TAg transgenic animals were examined, respectively, for the expression of lacZ (OSP-lacZ) or the development of tumors (OSP-TAg). RESULTS: The expression of lacZ in the resulting OSP-lacZ mice was restricted to the ovary as determined by X-gal staining of multiple organs. Immunohistochemical detection of beta-galactosidase showed lacZ expression mainly in the granulosa cells and ovarian surface epithelial cells. OSP-TAg mice developed tumors in a variety of tissues, including unilateral granulosa cell tumors in two of three female founder mice. In the contralateral ovary of one mouse with a granulosa cell tumor, there were alterations in the ovarian surface epithelial cells suggestive of preneoplasia. CONCLUSIONS: Although the OSP-1 promoter was able to restrict reporter gene expression to the ovary in transgenic mice, the expression of TAg in the OSP-TAg mice resulted in ovarian tumors as well as tumors in numerous other organs. This indicated that although transcription from the OSP-1 promoter occurs predominantly in the ovary, this promoter is sufficiently leaky in cells in other tissues to permit their tumorigenic conversion by SV40 TAg. PMID- 12759155 TI - Cannabis use and anxiety in daily life: a naturalistic investigation in a non clinical population. AB - The study's objective was to investigate in a non-clinical population the association between cannabis use and anxiety in daily life using the Experience Sampling Method (ESM). Seventy-nine subjects with high or low levels of cannabis use were selected among a sample of 685 undergraduate university students. ESM was used to collect information on cannabis use and state-anxiety in daily life. DSM-IV diagnoses were assessed using a structured clinical interview. Statistical analyses were performed using multilevel linear random regression models. There was no significant association between the level of state anxiety and cannabis use in daily life. However, a diagnosis of agoraphobia was significantly associated with increased likelihood of cannabis use, independent of state anxiety and other confounding factors. No evidence was found for an anxiolytic or anxiogenic effect of cannabis in daily life. This finding does not support the hypothesis that subjects with high levels of anxiety use cannabis as a means of self-medication. The association between agoraphobia and cannabis use in daily life may be explained by anticipatory anxiety secondary to previous cannabis induced panic-like symptoms. PMID- 12759156 TI - Aldehyde dehydrogenase 2 genotypes, alcohol flushing symptoms and drinking patterns in Thai men. AB - The drinking behavior, alcohol-induced facial flushing and ALDH2 genotypes were determined in 283 Thai men comprising 85 who were alcohol-dependent, 62 hazardous/harmful drinkers and 136 non-drinkers or infrequent drinkers. A structured interview questionnaire, containing the 'tri-level' method and the Alcohol-Use Disorders and Associated Disabilities Schedule, was used to determine the quantity of drinking and the number of alcohol-related adverse experiences. The study revealed the mutant ALDH2*2 allele in 44 (15.5%) subjects. The risks of being alcohol-dependent and of having hazardous/harmful drinking were lower in individuals with heterozygous ALDH2*1/*2, compared with homozygous ALDH2*1/*1 [relative probability ratios (95% CI) 0.14 (0.05-0.41) and 0.23 (0.08-0.61), respectively]. Eighty percent of those who were heterozygous and 28% of those who were homozygous ALDH2*1 reported flush symptoms after drinking alcohol. Twenty nine percent of homozygous ALDH2*1 individuals, but only 9% of heterozygous subjects, drank almost everyday (24-30 days/month). Similarly, higher percentages of people drinking beyond the safety limit (>60 g/day) and having alcohol-related problems were observed in homozygous ALDH2*1 compared with heterozygous individuals: 32% vs. 5% and 27% vs. 12%, respectively. Overall, the study supports the role of the mutant ALDH2*2 allele in preventing high alcohol consumption and the development of alcohol dependence in a Thai population. PMID- 12759158 TI - Positive association between panic disorder and polymorphism of the serotonin 2A receptor gene. AB - Family and twin studies have shown that genetic factors play an important role in the etiology of panic disorder. However, linkage and association studies using DNA markers have yielded inconclusive results. Increased serotonin neurotransmission may cause or be related to panic disorder. Assuming that genes regulating the serotonin system are involved in the pathogenesis of panic disorder, the authors searched for a genetic association of panic disorder with the serotonin 1A (HTR1A), 2A (HTR2A), and 2C (HTR2C) receptor genes. HTR1A, HTR2A and HTR2C polymorphisms were detected by the polymerase chain reaction method with analysis of restriction fragment-length polymorphisms (PCR-RFLP). The subjects were 63 biologically unrelated patients with panic disorder and 100 biologically unrelated normal control subjects who were native Japanese living in the western area of Japan. HTR1A and HTR2C showed no significant association with panic disorder. However, the frequency of the MspI A2 allele of HTR2A was significantly higher in the patients than in the normal control subjects. The study showed a positive association between panic disorder and the HTR2A gene, suggesting that HTR2A plays an important role in the pathogenesis of panic disorder. PMID- 12759157 TI - Possible association between serotonin transporter promoter region polymorphism and impulsivity in Koreans. AB - Serotonin has become the major focus of biological studies of suicidal behavior and impulsive-aggressive behavior in humans. The serotonin transporter (5-HTT) gene is one of the important genes involved in the regulation of serotonin transmission. We examined the association of impulsivity in Korean populations with a functional polymorphism of the promoter region of the 5-HTT gene (5 HTTLPR). We recruited 186 adolescent prisoners and 64 medical students. Impulsivity was measured using the Barratt Impulsiveness Scale and we divided all subjects into three groups: impulsive subjects (IS, N=121), non-impulsive subjects (NIS, N=115) and an intermediate group (excluded, N=14). The 5-HTTLPR genotype was determined by polymerase chain reaction. All subjects were Korean men unrelated to each other. There were no significant differences in the genotype frequency of 5-HTTLPR-S/S, S/L and -L/L between the two groups in the Korean population (IS vs. NIS: 47.9 vs. 61.7%; 43.0 vs. 32.2%; and 9.1 vs. 6.1%, respectively). However, there was a statistically significant difference in allelic frequency of 5-HTTLPR-S and 5-HTTLPR-L between the two groups in the Korean population (IS vs. NIS: 69.4 vs. 77.8%; and 30.6 vs. 22.2%, respectively. From our results, this 5-HTTLPR polymorphism appears to be a possible candidate gene for impulsivity in the Korean population. PMID- 12759159 TI - Serotonergic activity measured by platelet [3H]paroxetine binding in patients with eating disorders. AB - Most of the evidence from pharmacological studies supports the hypothesis of a serotonergic (5-HT) dysregulation in eating disorders (ED), though a specific alteration related to the major ED subtypes, anorexia (AN) and bulimia nervosa (BN), has not been identified yet, possibly because of changes over time in ED nosology. The aim of the present study was to verify whether differences in serotonergic activity, measured by platelet [3H]paroxetine binding, would validate current ED classification. Platelet [3H]paroxetine binding was investigated in 26 patients with eating disorders diagnosed in accord with DSM-IV criteria (AN, n=11; BN, n=15) and 26 normal weight controls of comparable age; ED symptomatology was assessed by the Diagnostic Schedule for Eating Disorders. ED patients had significantly lower B(max) values than controls (288.5+/-109.2 vs. 1396.8+/-251.3 fmol/mg), whereas the K(d) was not significantly altered (0.12+/ 0.13 and 0.12+/-0.05 nM, respectively). Among patients, differences in B(max) were related neither to DSM-IV subtypes nor to clinical variables such as presence of binge-eating, purging, impulsive behaviors, or symptoms of depression. Although ED patients share a dysregulation in serotonergic activity, DSM-IV subtype classification was not validated by [3H]paroxetine binding, and hence does not correspond to a specific 5-HT profile. PMID- 12759160 TI - Somatic symptoms in treatment-resistant depression. AB - The purpose of this work was to study the prevalence of somatic symptoms in patients with treatment-resistant depression (TRD) and their impact on the response to antidepressant treatment. Somatic symptoms were assessed during the screen visit with the Symptom Questionnaire (SQ) somatic symptom (SQ-SS) and somatic well-being sub-scales (SQ-SWB) in 40 patients with TRD enrolled in a 6 week open trial of nortriptyline. A logistic regression was used to test whether SQ-SS or SQ-SWB scores predicted clinical response to nortriptyline. Ninety-five percent of patients reported at least one somatic symptom. Higher SQ-SS scores during the screen visit predicted poorer response to nortriptyline. There was a trend for lower SQ-SWB scores to predict poor response to nortriptyline. None of the patients with SQ-SS scores above the mean for the sample responded to nortriptyline. The overwhelming majority of patients with TRD presented with somatic symptoms. In addition, a greater number of somatic symptoms during the screen visit placed patients at risk for further treatment resistance. PMID- 12759161 TI - Melatonin excretion with affect disorders over age 60. AB - Numerous studies have reported low melatonin secretion in depression, but other studies have suggested no deficit or an increase. Alterations of circadian phase or duration of melatonin secretion have also been described. Since melatonin secretion decreases as we age, it seemed interesting to examine melatonin and depression in an aging sample. Volunteers who complained of mood or sleep problems were recruited for studies in which fractional urine specimens were collected for 24 h, both at home and in the laboratory. The major metabolite, 6 sulfatoxymelatonin (aMT6s), was determined by radioimmunoassay. Of 72 volunteers aged 60-78 years, seven had current major depression and 55% had a lifetime history of an affective disorder. A 55-fold range of home aMT6s excretion rates was observed. A lifetime history of any affective disorder was significantly associated with greater log(10)[mesor] aMT6s excretion in home collections and laboratory collections, but current affective disorders were neither significantly related to melatonin excretion nor to aMT6s acrophase timing, onset, offset or duration. These results are only weakly consistent with a photoperiodic hypothesis of depression. PMID- 12759162 TI - Lymphocytes from bipolar and schizophrenic patients share common biochemical markers related to histone synthesis and histone cell membrane localization characteristic of an activated state. AB - In a previous communication, based on the total histone and histone variants' synthesis rates, biochemical parameters used for the characterization of the activation state of lymphocytes, we showed that a portion of the lymphocyte population obtained from peripheral blood of patients with bipolar disorder in the manic and/or depressed phases of the illness were in an activated state as opposed to normothymic patients and control subjects whose lymphocytes are in a resting, Go, state. In light of these previous findings, in the present investigation, we have analyzed total histone synthesis rates and the H2A and H3 histone variants' synthesis pattern of acid-extracted histones from the lymphocytes' nuclear fraction obtained from control subjects, patients with bipolar disorder in all phases of the illness, and patients with schizophrenia. Additional biochemical parameters, such as total cellular protein and DNA synthesis rates, were also studied. Moreover, recent findings from other investigators showed the association of histones on the plasma membrane fraction of PHA-activated, but not Go resting lymphocytes. Based on these results, acid extracted proteins from the plasma membrane fraction obtained from control, bipolar patients in all phases of the illness, and schizophrenic patients were analyzed by immunoblotting using a polyclonal histone antibody, anti-H2B. All biochemical parameters tested show that a portion of the lymphocyte population from bipolar, i.e. manic and depressive, as well as schizophrenic patients are in an activated state and clearly indicate that the unusual for lymphocytes cell cycle-related histone biochemical properties are common to both disorders. PMID- 12759163 TI - Increased sympathetic response to standing in panic disorder. AB - Although autonomic function has been investigated in panic disorder (PD), previous studies have not used non-invasive beat by beat blood pressure (BP) monitoring to assess the rapid dynamics of BP during autonomic reflex tests. The hypothesis of the current study was that patients with PD would show increased cardiovascular sympathetic reactivity compared with healthy or anxious controls, as assessed by the initial overshoot of diastolic BP during the immediate response to standing. Patients with PD (n=56), social phobia (n=28) and healthy volunteers (n=56) were tested using finger photoplethysmography during an orthostatic challenge. Panic disorder patients showed an increased BP overshoot compared with both control groups. Moreover, in a preliminary assessment of selective serotonin reuptake inhibitor treatment effects, the BP overshoot was significantly reduced towards normal values. These findings are consistent with recent evidence for increased sympathetic baroreflex function in PD and may be relevant to the pathophysiology of the disorder. PMID- 12759164 TI - Test-retest reliability of the Temperament and Character Inventory in patients with opiate dependence. AB - We examined the test-retest reliability of the Temperament and Character Inventory (TCI) in a clinical sample of 29 inpatients with opiate dependence disorder (DSM-IV). The previously validated French translation of the TCI was administered at baseline and again four weeks later. Intraclass correlation coefficients (ICCs) were used to estimate stability of the TCI over time for the 15 patients who completed the study. For all ICCs, the TCI showed satisfactory to excellent stability across all factors (ICC= 0.66-0.82). Stability was lower for the two temperamental traits of 'persistence' (ICC=0.51) and 'reward dependence' (ICC=0.63), possibly reflecting both clinical instability and measurement errors. These results highlighted the overall stability of the TCI in patients with opiate dependence and provided evidence for the usefulness of this questionnaire, which was originally designed to explore genetic and environmental factors underlying normal and abnormal personality dimensions. Further studies are required to confirm these results on larger clinical samples. PMID- 12759165 TI - How does the Hospital and Anxiety and Depression Scale measure anxiety and depression in healthy subjects? AB - Many anxiety and depression scales are commonly used, although the assumption that they all measure the same construct may be questioned. Thus, researchers have to pay attention to the nature of the scales they use. The Hospital Anxiety and Depression Scale (HADS) was constructed in 1983 to allow a rapid and separate measure of depression and generalised anxiety. Surprisingly, since its introduction, there has been no comprehensive documentation of its psychometric properties. Therefore, as a contribution to assessing the construct validity of the HADS, we conducted a set of confirmatory factor analyses in a sample of 195 healthy students. None of the formerly proposed models fit our data. We were able to split the original Anxiety subscale into two components that we have labelled 'Anxiety' and 'Restlessness', while the original Depression subscale is slightly modified. The results are discussed from both clinical and theoretical points of view. PMID- 12759166 TI - Modular concepts of brain organization and the neuropathology of psychiatric conditions. PMID- 12759167 TI - Altered glutamate uptake in peripheral tissues from Down syndrome patients. AB - Overexpression of APP and SOD induces beta-amyloid deposition and oxidative stress in Down syndrome (DS) patients. Both phenomena may impair glutamate transport and decreased glutamate uptake sites have been demonstrated in patient brains at autopsy. Since alterations of APP metabolism and oxidative damage are systemic, we investigated glutamate uptake in platelets and fibroblasts from DS patients to explore whether abnormalities in this process are inherent properties of DS cells and not secondary to neurodegeneration. Glutamate uptake was significantly decreased in platelets (P<0.005 vs. control) and fibroblasts (P<0.001 vs. control) from DS patients, particularly in those with free trisomy and with mitochondrial point mutations. Systemic impairment of glutamate uptake in DS is suggested, probably related to APP overexpression and mitochondrial dysfunction. Such mechanisms may contribute to neurodegeneration and dementia development in these patients. PMID- 12759168 TI - Vacuole-creating protein in neurodegenerative diseases in humans. AB - Vacuole-creating protein (VCP) is a member of the ATPases associated with diverse cellular activities and is a putative sensor protein for degenerative proteins. Immunohistochemical examinations demonstrated that VCP was observed in ubiquitin positive intraneuronal inclusions in motor neuron disease with dementia, ballooned neurons in Creutzfeldt-Jakob disease, dystrophic neurites of senile plaque in Alzheimer's disease, and Lewy and Marinesco bodies and Lewy neurites in Parkinson's disease, while granules of granulovacuolar degeneration and neurofibrillary tangles in Alzheimer's disease were not positively stained for VCP. These results indicate that VCP reacts with abnormal or misfolded proteins and plays a role in accelerating the process of degeneration and cell death. The elucidation of an association between VCP and these degenerative proteins will provide an important clue for understanding common mechanisms underlying neurodegenerative diseases. PMID- 12759169 TI - The AMPA receptor antagonist NBQX prolongs survival in a transgenic mouse model of amyotrophic lateral sclerosis. AB - alpha-Amino-3-hydroxy-5-methylisoxazole propionic acid (AMPA) receptor-mediated excitotoxicity has been implicated in the selective motor neuron loss in amyotrophic lateral sclerosis (ALS). The extent to which excitotoxicity contributes to motor neuron death remains incompletely understood. We therefore tested the potent and selective AMPA/kainate receptor antagonist 1,2,3,4 tetrahydro-6-nitro-2,3-dioxo-benzo[f]quinoxaline-7-sulfonamide (NBQX) on motor neurons in culture and in the G93A mouse model for familial ALS. Kainate-induced currents and changes in intracellular Ca(2+) concentration were measured with the perforated patch clamp technique combined with Ca(2+) imaging, motor neuron death was quantified by counting experiments and G93A mice were treated with saline or 8 mg/kg NBQX. NBQX blocked kainate-induced currents and concomitant changes in intracellular Ca(2+), prevented the kainate-induced motor neuron death, and prolonged survival of G93A mice. PMID- 12759170 TI - Electrical and pharmacological properties of the slow depolarization induced by Schaffer collateral stimulation in astrocytes of the rat hippocampal CA1 region. AB - Intracellular recordings were made from astrocytes of the hippocampal CA1 region in rat slice preparations. A single stimulus at the Schaffer collaterals with high intensities (>/=10 V), but not low intensities (<10 V), induced a slow depolarization similar to the evoked postsynaptic potentials observed in hippocampal CA1 neurons. The slow depolarization was almost abolished in the presence of alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate/kainate receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (20 microM) and N-methyl-D aspartate receptor antagonist DL-2-amino-5-phosphonopentanoic acid (100 microM). Tetrodotoxin (0.5 microM) or Co(2+) (2 mM) also abolished this slow depolarization. The slow depolarization reversed its polarity at -8 mV in the presence of tetraethylammonium (20 mM). These results suggest that the synaptically released glutamate (Glu) induces depolarization, which is mainly mediated by ionotropic Glu receptors, in astrocytes in the hippocampal CA1 region. PMID- 12759171 TI - Spinal sensitization mechanism in vincristine-induced hyperalgesia in mice. AB - Our aim was to investigate the possible involvement of spinal voltage-dependent Ca(2+) channels (VDCCs) in vincristine-induced hyperalgesia and also to characterize this hyperalgesic state in the spinal cord. Mice receiving vincristine displayed significantly lower mechanical nociceptive thresholds than controls. Intrathecal omega-conotoxin GVIA (an N-type blocker) produced dose dependent inhibition of the mechanical nociception, its antinociceptive effect being greater in vincristine-treated mice than in controls. The antinociception of omega-agatoxin IVA (a P/Q-type blocker) and calciseptine (an L-type blocker) were both slightly, but not significantly greater in vincristine-treated mice. An N-methyl-D-aspartate-receptor antagonist but not a tachykinin-NK1-receptor antagonist produced greater antinociception in vincristine-treated mice. These results suggest that vincristine-induced hyperalgesia involves a sensitization of the spinal processing of mechanical sensory information via a mechanism involving N-type but not P/Q- or L-type VDCCs. A spinal glutamatergic pathway also appears to be involved in this hyperalgesia. PMID- 12759172 TI - Lack of association between the interleukin-1alpha gene C(-889)T polymorphism and Alzheimer's disease in a Chinese population. AB - Interleukin 1A (IL-1A), a potent proinflammatory cytokine, has been implicated in the pathogenesis of Alzheimer's disease (AD). Several recent studies have shown that a polymorphism in the IL-1A locus -889 was associated with AD risk and onset age, however, other studies were not able to replicate these findings. In this investigation, the IL-1A C(-889)T polymorphism has been analyzed for a Chinese population of 234 AD patients and 170 controls. There were no significance differences in IL-1A genotype or allele frequencies comparing the AD cases and controls, even after stratification for age of onset and adjustment for apolipoprotein E genotype. The results reveal that it is not likely that the IL 1A C(-889)T polymorphism is involved in AD pathogenesis in the Chinese population. Further studies of the associations between other IL-1A genetic polymorphisms and AD are needed to fully elaborate the involvement of this gene in AD. PMID- 12759173 TI - Systemic oxytocin treatment modulates glucocorticoid and mineralocorticoid receptor mRNA in the rat hippocampus. AB - The present study investigated the effects of oxytocin treatment on hippocampal glucocorticoid receptors. Oxytocin (1 mg/kg s.c.) was administered once a day for 5 days to male Sprague-Dawley rats. The animals were sacrificed 1 day after treatment and expression of glucocorticoid and mineralocorticoid receptor (GR and MR) mRNA in the dorsal hippocampus was measured with in situ hybridization. The oxytocin treatment decreased GR mRNA expression in CA1+2 and the dentate gyrus (P<0.05), tended to decrease GR mRNA expression in CA3 (P=0.07), and increased MR mRNA expression in the dentate gyrus (P<0.05). These findings demonstrate that systemic oxytocin treatment induces changes in the hippocampal glucocorticoid receptors. Thus, oxytocin may modulate the activity of the hypothalamic-pituitary adrenal-axis also at the hippocampal level in rats. PMID- 12759174 TI - Nitric oxide-induced inhibition on striatal cells and excitation on globus pallidus neurons: a microiontophoretic study in the rat. AB - Single units were recorded in the striatum and in the globus pallidus (GP) of urethane-anesthetized rats under microiontophoretic administration of either Nomega-nitro-L-arginine methyl ester (L-NAME, inhibitor of nitric oxide synthase), or 3-morpholino-sydnonimin-hydrocloride (SIN-1, nitric oxide, NO donor). A steady baseline firing of sporadically discharging striatal neurons (basal firing rate <0.1 spikes/s) was evoked by a pulsed microiontophoretic ejection of glutamate. On striatal neurons, microiontophoretic application of SIN 1 induced a current-dependent inhibition (11/13), whereas L-NAME administration produced a clear excitation (9/9). On GP cells, the administration of SIN-1 had excitatory effects (10/15), whereas the administration of L-NAME reduced the neuronal activity (6/6). We hypothesize that NO could exert an intrinsic regulatory action on the activity of both striatal and GP cells. PMID- 12759175 TI - Postdecentralization plasticity of voltage-gated Na+ currents in rat glandular sympathetic neurons. AB - The kinetic properties of voltage-gated Na(+) currents in two groups of glandular postganglionic sympathetic neurons were assessed. The first group of neurons remained innervated by preganglionic axons until the day of current recordings, while the second--decentralized 4 weeks prior to recordings. An increase of maximum current amplitude and density was noted in decentralized neurons. Na(+) currents activated and time-dependently inactivated more slowly in decentralized than in control neurons. Furthermore, after decentralization the currents steady state inactivated at less hyperpolarized potentials as well as reactivated faster from inactivation. We conclude that the Na(+) currents in decentralized postganglionic glandular sympathetic neurons undergo up-regulation. PMID- 12759176 TI - Melatonin enhances lymphocyte proliferation and decreases the release of pituitary pro-opiomelanocortin-derived peptides in surgically traumatized rats. AB - The present study was undertaken to evaluate the effects of melatonin (MT) on spleen lymphocyte proliferation and release of pituitary pro-opiomelanocortin (POMC)-derived peptides in surgically traumatized rats. MT prevented the depression of lymphocyte proliferation induced by trauma in vivo and in vitro, and prevented the decrease of beta-endorphin and adrenocorticotropin in the pituitary in vivo, but dose-dependently inhibited the release of POMC-derived peptides from the pituitary in vitro. The culture media of the pituitaries derived from the traumatized rats inhibited lymphocyte proliferation of normal rats. These results suggest that MT can improve the trauma-induced depression of lymphocyte proliferation and inhibit the release of POMC-derived peptides. The neuroimmunomodulatory role of MT and POMC-derived peptides deserves further study. PMID- 12759177 TI - Effects of propofol on the activity of rat glutamate transporter type 3 expressed in Xenopus oocytes: the role of protein kinase C. AB - We investigated the effects of propofol on one type of glutamate transporter, excitatory amino acid transporter 3 (EAAT3) and the role of protein kinase C (PKC) in mediating these effects. Rat EAAT3 was expressed in Xenopus oocytes. L glutamate (30 microM)-induced membrane currents were measured. Propofol increased glutamate-induced inward currents significantly at two tested concentrations (30 and 100 microM) but not at other concentrations. Propofol (30 microM) significantly increased V(max), but not K(m) of EAAT3 for glutamate. The combination of phorbol-12-myrisate-13-acetate (PMA, a PKC activator) and propofol did not increase the responses further compared with PMA or propofol alone. Three PKC inhibitors (staurosporine, calphostin C, and chelerythrine) did not affect basal EAAT3 activity but significantly inhibited the propofol-enhanced EAAT3 activity. Our results suggest that propofol enhances EAAT3 activity at clinically relevant concentrations and PKC may mediate these effects. PMID- 12759178 TI - Failure to find causal mutations in the GABA(A)-receptor gamma2 subunit (GABRG2) gene in Japanese febrile seizure patients. AB - Recently, mutations in the GABA(A)-receptor gamma2 subunit (GABRG2) gene were identified in two families with generalized epilepsy with febrile seizures plus (GEFS+) and two families with childhood absence epilepsy (CAE) and febrile seizures (FS). We tested the hypothesis that genetic variations in the GABRG2 gene confer susceptibility to FS in the Japanese population. We performed a systematic search for mutations in 94 unrelated Japanese patients with FS and detected six variants (-158C>T, 315C>T, 588T>C, IVS5-55C>T, IVS7+20G>A, and IVS7 141T>A). No non-synonymous mutation was detected. We genotyped three exonic polymorphisms and performed a case control study and a transmission disequilibrium test using 55 independent complete trios with FS and 106 control subjects. None of these polymorphic alleles were significantly associated with FS. Our results indicate that genomic variations of GABRG2 are not likely to be substantially involved in the etiology of FS in the Japanese population. PMID- 12759179 TI - Widely integrative properties of layer 5 pyramidal cells support a role for processing of extralaminar synaptic inputs in rat neocortex. AB - We have compared the length, strength, conduction velocity and divergence of horizontal connections onto layers 2/3 and 5 neurons in slices of rat primary somatosensory neocortex. Slices were cut along laminar borders to eliminate most vertical connections, and excitatory postsynaptic potentials (EPSPs) were recorded from pyramidal cells in adjacent uncut tissue. When electric stimuli were delivered within the same layer as the recorded cell, EPSPs could be evoked up to 2000 microm away for both layers 2/3 and 5 pyramids. Estimates of horizontal axonal conduction velocities ( approximately 0.4 m/s) and the thresholds for activation also did not differ between layers. However, layers 2/3 cells rarely responded to stimuli delivered to isolated deeper layers, while layer 5 neurons were often excited by horizontal inputs from isolated layers 2/3 and 4. PMID- 12759180 TI - Suppression of postsynaptic density protein 95 expression attenuates increased tyrosine phosphorylation of NR2A subunits of N-methyl-D-aspartate receptors and interactions of Src and Fyn with NR2A after transient brain ischemia in rat hippocampus. AB - The effects of suppression of postsynaptic density protein 95 (PSD-95) expression on the increased tyrosine phosphorylation of N-methyl-D-aspartate receptor subunit NR2A and interactions of Src and Fyn with NR2A after brain ischemia were investigated by immunoprecipitation and immunoblotting. Transient (15 min) brain ischemia was induced by the four-vessel occlusion method in Sprague-Dawley rats. Intracerebroventricular infusion of PSD-95 antisense oligonucleotides (every 24 h for 3 days before ischemia), but not missense oligonucleotides or vehicle, not only markedly decreased the protein level of PSD-95 but also attenuated the elevated tyrosine phosphorylation of NR2A and interactions of Src and Fyn with NR2A induced by 6 h of reperfusion following ischemia in the hippocampus. The protein levels of NR2A, Src and Fyn had no differences under the above conditions. These data suggested that PSD-95 is critical for facilitating NR2A tyrosine phosphorylation by Src family kinases in postischemic brain. PMID- 12759181 TI - Human neural stem cells can migrate, differentiate, and integrate after intravenous transplantation in adult rats with transient forebrain ischemia. AB - Intraparenchymally transplanted rodent-origin neural and human-origin mesenchymal stem cells migrate and differentiate in neurological diseases. By intravenously injecting human neural stem cells, we showed that transplanted human neural stem cells migrate to the damaged hippocampus, proliferate and differentiate into mature neurons and astrocytes in the adult rat brain with transient forebrain ischemia. We also demonstrated the migratory course of implanted human neural stem cells after intravenous injection. Our findings show that transplanted human neural stem cells differentiate into mature neurons to replace lost neural cells in the adult hippocampus with human-rat neural chimeras. PMID- 12759182 TI - Identification of nuclear/nucleolar localization signal in Aplysia learning associated protein of slug with a molecular mass of 18 kDa homologous protein. AB - We isolated a learning associated protein of slug with a molecular mass of 18 kDa (LAPS18) homologue from the expressed sequence tag database of Aplysia kurodai and named it Aplysia LAPS18-like protein (ApLLP). ApLLP encodes 120 amino acids and has 57% identity with LAPS18. To examine the subcellular expression pattern of ApLLP we constructed an EGFP-tagged ApLLP fusion protein and overexpressed it in both Aplysia neurons and COS-7 cells. In contrast to the previous findings, which showed that LAPS18 is secreted by COS-7 cells, ApLLP-EGFP was localized to the nucleus, and most of it to nucleoli. Analysis of deletion mutants of ApLLP EGFP showed that the N-terminal and the C-terminal nucleolar and nucleus localization signal sequences are important for localization to the nucleus and the nucleoli. PMID- 12759183 TI - 1,25-dihydroxyvitamin D3 induces nerve growth factor, promotes neurite outgrowth and inhibits mitosis in embryonic rat hippocampal neurons. AB - There is an accumulation of evidence implicating a role for vitamin D(3) in the developing brain. The receptor for this seco-steroid is expressed in both neurons and glial cells, it induces nerve growth factor (NGF) and it is a potent inhibitor of mitosis and promoter of differentiation in numerous cells. We have therefore assessed the direct effect of vitamin D(3) on mitosis, neurite outgrowth, as well as NGF production as a possible mediator of those effects, in developing neurons. Using cultured embryonic hippocampal cells and explants we found the addition of vitamin D(3) significantly decreases the percentage of cultured hippocampal cells undergoing mitosis in conjunction with increases in both neurite outgrowth and NGF production. The role of vitamin D(3) during brain development warrants closer scrutiny. PMID- 12759184 TI - Response of neuropathic trigeminal pain to the combination of low-dose nalbuphine plus naloxone in humans. AB - We report on the response of medically refractory neuropathic trigeminal pain in three patients to intravenous administration of a combination of the kappa partial agonist opioid nalbuphine and the opioid antagonist naloxone. Each of the three patients had developed a painful peripheral neuropathy as a complication of chemical or mechanical injury to the trigeminal nerve. Each patient had been tried on a number of analgesics, including mu-opioids, and had not gained relief or was not able to tolerate side effects of the medications. Pain intensity was measured for 3 h following drug administration using a 10 cm visual analog scale. All three patients reported marked decrease in pain following administration of the nalbuphine and naloxone combination. These findings suggest a novel approach to the management for neuropathic pain. PMID- 12759185 TI - Nitrous oxide reverses the increase in striatal dopamine release produced by N methyl-D-aspartate infusion in the substantia nigra pars compacta in rats. AB - Bilateral administration of NMDA (5 x 10(-10) mol) in the substantia nigra pars compacta increases the striatal dopamine (DA) release. However, this enhancing effect of NMDA was suppressed by nitrous oxide exposure at 0.1 MPa, which induced per se a decrease of the DA release. These results show that nitrous oxide exerts a reversal effect on the increase in striatal DA release produced by NMDA receptor activation in the substantia nigra pars compacta. This observation may be related to the fact that nitrous oxide is thought to produce its effects by acting as an NMDA receptor antagonist. PMID- 12759186 TI - Glutamine is incorporated at the nonsense codons UAG and UAA in a suppressor-free Escherichia coli strain. AB - Readthrough of the nonsense codons UAG, UAA, and UGA is seen in Escherichia coli strains lacking tRNA suppressors. Earlier results indicate that UGA is miscoded by tRNA(Trp). It has also been shown that tRNA(Tyr) and tRNA(Gln) are involved in UAG and UAA decoding in several eukaryotic viruses as well as in yeast. Here we have investigated which amino acid(s) is inserted in response to the nonsense codons UAG and UAA in E. coli. To do this, the stop codon in question was introduced into the staphylococcal protein A gene. Protein A binds to IgG, which facilitates purification of the readthrough product. We have shown that the stop codons UAG and UAA direct insertion of glutamine, indicating that tRNA(Gln) can read the two codons. We have also confirmed that tryptophan is inserted in response to UGA, suggesting that it is read by tRNA(Trp). PMID- 12759187 TI - Pig whey acidic protein gene is surrounded by two ubiquitously expressed genes. AB - A 140-kb pig DNA fragment containing the whey acidic protein (WAP) gene cloned in a bacterial artificial chromosome (BAC344H5) has been shown to contain all of the cis-elements necessary for position-independent, copy-dependent and tissue specific expression in transgenic mice. The insert from this BAC was sequenced. This revealed the presence of two other genes with quite different expression patterns in pig tissues and in transfected HC11 mouse mammary cells. The RAMP3 gene is located 15 kb upstream of the WAP gene in reverse orientation. The CPR2 gene is located 5 kb downstream of the WAP gene in the same orientation. The same locus organization was found in the human genome. The region between RAMP3 and CPR2 in the human genome contains a WAP gene-like sequence with several points of mutation which may account for the absence of WAP from human milk. PMID- 12759188 TI - "Blue heart": characterization of a mifepristone-dependent system for conditional gene expression in genetically modified animals. AB - A detailed characterization of a cardiac muscle-specific, ligand-regulated gene expression system was performed in transgenic mice using the inducing ligand mifepristone (MFP). Several lines of double transgenic mice were created that expressed a bacterial lacZ reporter gene in the heart, under the control of a MFP activated transcription factor constitutively expressed in cardiac muscle. The transgenic mice, which were administered MFP at a dose of 1 micromol/l in the drinking water, responded to the ligand within 24 h. Induction of beta galactosidase enzyme activity in the heart continued for up to 21 days and resulted in an average 17-fold increase in enzyme activity. The highest individual animal response measured was a 94-fold increase in enzyme activity. The EC(50) for MFP induction of beta-galactosidase activity in the heart was 0.7 micromol/l when MFP was administered in the drinking water. Pharmacokinetic analysis of MFP dosing in wild-type FVB/N mice showed that absorption was very rapid (T(max) 1-10 min), bioavailability was modest ( approximately 10%) and the t(1/2) of MFP in mouse plasma was determined to be approximately 5 h. Thus, the system functions effectively in transgenic mouse heart where induction of gene expression is sensitive and can be accomplished by a simple and broadly applicable drinking water protocol. PMID- 12759189 TI - 3' Splicing variants of ret receptor tyrosine kinase are differentially expressed in mouse embryos and in adult mice. AB - The RET protooncogene encodes for a transmembrane receptor tyrosine kinase and plays a crucial role in nephrogenesis and the enteric nervous system (ENS) development. Alternative splicing at the 3' end of the RET gene generates 3' splicing variants that encode RET 9, RET 51 and RET 43 isoforms. It has been hypothesized that these isoforms perform distinct functions and that their expressions are differentially regulated during mammalian development. To gain an insight into the expression patterns of various ret isoforms during embryogenesis, we investigate the temporal and spatial expressions of ret gene in mouse embryos and in adult mice. We characterized the 3' end of the mouse ret gene and localized the alternatively spliced exons. Using 3' rapid amplification of cDNA ends (3' RACE) and reverse transcription-polymerase chain reaction (RT PCR), ret 9 and ret 51 transcripts were identified in both mouse embryos and adult mouse tissues. However, the ret 43 transcript was not. Using in situ hybridization, we showed that ret 9 was the dominant ret encoding transcript in mouse embryos. Transcripts of ret 9 were detected in all cranial ganglia; in the sensory and autonomic ganglia of the trunk; in a subset of neurons of the dorsal root ganglion (DRG); in the motor neurons of the spinal cord; in the developing lung and excretory systems; in the enteric neuroblasts of the ENS; and in the thyroid lobes. In contrast, ret 51 expression was weak and restricted to the motor column of the spinal cord, the DRG, the enteric neuroblasts, the lung bud and the kidney. In adult mice, ret 9 expression was relatively widespread in many organs while that of ret 51 was rather restricted. Our data indicated that ret isoforms are temporally and spatially regulated in mouse embryos and adult mouse. PMID- 12759190 TI - Positioning of mRNA codons with respect to 18S rRNA at the P and E sites of human ribosome. AB - Positioning of each nucleotide of the E site and the P site bound codons with respect to the 18S rRNA on the human ribosome was studied by cross-linking with mRNA analogs, derivatives of the hexaribonucleotide UUUGUU (comprising Phe and Val codons) that carried a perfluorophenylazide group on the second or the third uracil, and a derivative of the dodecaribonucleotide UUAGUAUUUAUU with a similar group on the guanine residue. The location of the modified nucleotides at any mRNA position from -3 to +3 (position +1 corresponds to the 5' nucleotide of the P site bound codon) was adjusted by the cognate tRNAs. A modified uridine at positions from -1 to +3 cross-linked to nucleotide G1207 of the 18S rRNA, and to nucleotide G961 when it was in position -2. A modified guanosine cross-linked to nucleotide G1207 if it was in position -3 of the mRNA. These data indicate that nucleotide G961 of the 18S rRNA is close only to mRNA positions -3 and -2, while G1207 is in the vicinity of positions from -3 to +3. The latter suggests that there is a sharp turn between the P and E site bound codons that brings nucleotide G1207 of the 18S rRNA close to each nucleotide of these codons. This correlates well with X-ray crystallographic data on bacterial ribosomes, indicating existence of a sharp turn between the P site and E site bound codons near a conserved nucleotide G926 of the 16S rRNA (corresponding to G1207 in 18S rRNA) close to helix 23b containing the conserved nucleotide 693 of the 16S rRNA (corresponding exactly to G961 of the 18S rRNA). PMID- 12759191 TI - Coprinus cinereus DNA ligase I during meiotic development. AB - DNA ligase I is thought to be essential for DNA replication, repair and recombination, at least in the mitotic cell cycle, but whether this is also the case during the meiotic cell cycle is still obscure. To investigate the role of DNA ligase I during the meiotic cell cycle, we cloned the Coprinus cinereus DNA ligase I cDNA (CcLIG1). Northern blotting analysis indicated that CcLIG1 is expressed not only in the premeiotic S-phase but also during the meiotic cell cycle itself. Especially, intense signals were observed in the leptotene and zygotene stages. Western blotting analysis indicated that CcLIG1 is expressed through the meiotic cell cycle and immunofluorescence also showed CcLIG1 protein staining in meiotic cells. Interestingly, the patterns was similar to that for the C. cinereus proliferating cell nuclear antigen gene (CcPCNA) and immunoprecipitation analysis suggested that CcPCNA binds to CcLIG1 in crude extracts of meiotic prophase I tissues. Based on these observations, relationships and roles during the meiotic cell cycle are discussed. PMID- 12759192 TI - Swine Toll-like receptor 9(1) recognizes CpG motifs of human cell stimulant. AB - Complementary DNA (cDNA) encoding swine Toll-like receptor 9 (sTLR9) was isolated from Peyer's patches (Pps) of gut-associated lymphoid tissue (GALT). The complete open reading frame (ORF) of sTLR9 contains 3093 bp coding deduced 1030 amino acid residues. The amino acid sequence of sTLR9 was characterized by a signal peptide followed by multiple leucine-rich repeats, a transmembrane sequence and a cytoplasmic domain homologous to that of the human interleukin-1 receptor (TIR). The sTLR9 showed a higher amino acid identity with humans (81.8%) and felis catus (86.7%) than mice (74.9%). The HEK293T cells transfected with pCXN2.1-FLAG DNA containing the sTLR9 cDNA were expressed sTLR9 as a membrane-bound molecules, which were reactive with anti-sTLR9 rabbit polyclonal antibody. Moreover, the transfectant was responsible for the CpG oligo DNA. sTLR9 was preferentially expressed in Pps and mesenteric lymph nodes (MLNs), and its degree was approximately three times higher than a spleen but weak in the other tissues by the real-time quantitative PCR analyses. The strong expression of sTLR9 in Pps and MLNs and its recognizing CpG DNA for human cell stimulant are shown first in this study, which may help in understanding the intestinal immune system mediated by a bacterial DNA through TLR9. PMID- 12759193 TI - Conference on abuse liability assessment of CNS drugs. PMID- 12759194 TI - Keeping the lid on: a century of drug regulation and control. AB - Since the early 1900s, national and international drug control legislation has acted as a key site of contention between important societal actors. Physicians and pharmacists, regulators and drug companies, patients and addicts, and researchers and pharmacologists all attempted to influence formulation and interpretation of the rules that regulate access to addicting but medically useful substances. The 1970 Controlled Substances Act (CSA) consolidated and rationalized previous US domestic legislation and paid careful attention to the international aspects of the issue. Yet the CSA also incorporated long-standing fundamental disputes about who would act as gatekeepers, what criteria would be employed in regulatory decisions, and the basic goals of drug control legislation. Rather than view the CSA as a beginning or an end, it is better conceived as a major milepost in a century-long odyssey of maneuvering among interested parties for advantage in a complex regulatory environment. Instead of providing a definitive authoritative structure to which all parties must adhere, the CSA has served as a vehicle for discernment and continuous renegotiation of essential concepts such as "abuse liability". PMID- 12759195 TI - Guidelines and methodological reviews concerning drug abuse liability assessment. AB - Regulatory control of drugs with abuse liability is an important component of drug control policy and is believed to help prevent nonmedical use. To be maximally effective, this requires a scientific assessment of abuse liability of drugs considered for regulatory control. These assessments have relied extensively on laboratory-based animal and human testing, but also utilize information from clinical trials, actual abuse and other sources. Here, we discuss recommendations and guidelines that have been proposed for abuse liability assessment and describe important review papers and conference proceedings that have addressed this matter, focusing primarily on drugs with medical usefulness. Historically, there is substantial consensus about how to approach abuse liability evaluation of drugs with actions similar to those of abused opiates, stimulants, depressants, and to a somewhat lesser extent, cannabinoids and hallucinogens, and much of what has been recommended for abuse potential assessment in the past remains valid and useful. On the other hand, novel CNS-active medications which cannot be readily classified with these traditional drugs of abuse are increasingly under development. In addition, advances in the science of abuse liability assessment need to be incorporated into future guidelines and recommendations on this subject. Developers of new medications need guidance on how to utilize scientific research to maximize therapeutic benefit while minimizing risk for abuse. Thus, another goal of this review has been to identify areas where critical thinking and new guideline development are needed. PMID- 12759196 TI - Principles of initial experimental drug abuse liability assessment in humans. AB - This paper describes the rationale and procedures for conducting what is considered by many to be the current "gold standard" for initial abuse liability testing of a novel compound: the classic acute dose-effect comparison study in volunteers with histories of drug abuse. Such a trial is most appropriate for predicting the likelihood of abuse by drug abusers and, in turn, the extent of drug diversion and illicit street sales if the novel compound became available in the community. The dose-effect abuse liability trial typically involves a double blind complete crossover design in 10-14 subjects with histories of polydrug abuse in a controlled clinical pharmacology laboratory setting. Drug conditions usually involve placebo, three doses of the novel compound and three doses of an appropriate reference compound of known abuse liability. In each session, the time-course of effects of a single drug dose are evaluated. Intervals between experimental sessions are typically 1 to several days. The importance of testing high supra-therapeutic doses of the novel drug for the validity of the trial is emphasized, and the use of a dose run-up pilot study for selecting maximal doses and matching doses between the novel and comparison compound is explained. The rationale and description of outcome measures is discussed, including measures that reflect likelihood of abuse (e.g. drug vs. money choice and subject ratings of liking, good effects, estimated monetary street value), secondary measures that should be considered in interpreting likelihood of abuse (e.g. drug identification, subject-rated side effects and mood changes), and additional concurrent measures to establish equivalence of the novel and comparison compound (e.g. behavioral performance, observer-rated assessments, physiological measures). PMID- 12759197 TI - Principles of drug abuse liability assessment in laboratory animals. AB - This paper describes the rationale for use of preclinical assessments of abuse liability in laboratory animals, and then discusses "cross-cutting" methodological issues that apply to behavioral evaluations intended to contribute to an abuse liability evaluation package. Issues include use of: (1) positive and negative control conditions; (2) full dose-effect evaluations, (3) multiple dependent measures, (4) pharmacokinetic evaluations to guide choice of dose ranges, (5) a species for which good methodological and comparative data are available to aid interpretation of results, and (6) appropriate methods for the group or single-subject experimental design selected. The remainder of the paper describes basic methodology by which three core pieces of behavioral data required by the Food and Drug Administration for its use in the overall abuse liability analysis can be obtained preclinically. Reinforcing effects are assessed in study of drug self-administration; drug discrimination assesses degree of overlap of interoceptive stimulus effects with relevant comparison drugs; physical dependence potential is determined by assessing whether a withdrawal syndrome occurs after chronic drug administration. Background and methodological issues specific to each procedure are discussed. A key consideration for cross-cutting and specific methodological issues is that choices made enable confident interpretation of both positive and negative results. PMID- 12759198 TI - Incorporating the assessment of abuse liability into the drug discovery and development process. AB - Evaluation of abuse liability is one of many obligations incurred by industrial sponsors in the development of medications acting on substrates in the central nervous system. In addition to providing the information necessary for a scheduling recommendation in the marketing application, the abuse liability assessment allows sponsors to estimate safety and commercial risks associated with scheduling, as well as to tailor their pre- and post-approval programs to collect information relevant to product misuse, illicit diversion and physical dependence. There are several important factors to consider before embarking on an abuse liability assessment, including the compound's primary and secondary biochemical activities, its absorption and metabolism, its final formulation, and its intended clinical population. Each of these factors will temper the timing and extent of the abuse liability program in animals and humans. Although every drug development program is unique in some way, a decision-making process may be applied to abuse liability assessment that will serve to better utilize limited resources and inform decisions regarding subsequent steps in the process. The emerging properties of the product will define the unique procedures best applied to assess it. PMID- 12759199 TI - Assessing abuse liability in clinical trials. AB - In this article, the use of data collected in registration-focused clinical trials to provide information concerning abuse liability of compounds under development is discussed. Registration-focused trials are limited by the small and select sample chosen for study participation and design constraints. However, most compounds under development are first administered to humans during the conduct of registration-focused trials, so this presents an opportunity to collect potentially important information about the effects of drugs in humans. At present, information concerning subjective effects and symptoms associated with drug discontinuation are not collected systematically. Reports are generally considered as adverse events (AEs) and are recorded on the case report forms (CRFs) in the investigators own language. There is generally no rating of drug "liking". The authors suggest strategies that could be implemented in registration-focused clinical trials to improve the information gathered with regard to subjective effects, abuse liability and discontinuation-emergent symptoms. Importantly, there remains much groundwork to be done in developing and validating appropriate assessment instruments and determining "threshold" levels for concern. Under the best of circumstances, registration-focused clinical trials have limited potential to detect abuse liability because of the small number of patients seen and the exclusion of many subjects who might be particularly vulnerable to the abuse of marketed compounds (i.e. individuals with substance use disorders). In cases where there are reasons to suspect that a drug under development has abuse potential, systematic exploration with a series of studies specifically designed to assess abuse potential must be conducted. PMID- 12759200 TI - Postmarketing surveillance for drug abuse. AB - Assessing actual abuse of prescribed medications requires postmarketing surveillance. In this article we discuss general systems of postmarketing surveillance that exist as of the end of 2002 in the United States and two medication-specific surveillance systems that were devised and tested. The two specific surveillance systems are compared with limitations highlighted. Postmarketing surveillance is in its infancy and requires more research on ways to improve its validity without inducing illicit experimentation. Information on comparator medications is highly recommended both to validate the system and to place the results in context. PMID- 12759201 TI - Abuse liability assessment of CNS drugs: conclusions, recommendations, and research priorities. PMID- 12759202 TI - Extraanatomical coronary artery bypass grafts on the beating heart for management of the severely atherosclerotic ascending aorta. AB - BACKGROUND: Crossclamping a severely atherosclerotic ascending aorta carries a significant risk of stroke in coronary artery bypass grafting. Besides other techniques aortic no touch concepts are increasingly applied for management of this problem. METHODS: Out of 407 patients undergoing epiaortic scanning during coronary artery bypass grafting 38 (9.3%) exhibited severe ascending aortic atherosclerosis. 22 of these patients (18 male, 4 female, age 72 (57-79) years, Parsonnet Score 11 (0-18), Euro Score 8 (2-13), McSPI Stroke Risk Index 6 (1-30) %) were operated on using a beating heart and aortic no touch technique. All patients received at least one internal mammary artery (IMA) in situ graft and additional extraanatomical bypass conduits: venous Y-graft from the IMA (n=14), arterial Y-graft from the IMA (n=3), vein graft from the axillary artery (n=3), vein graft from the IMA stump (n=2), vein graft from the innominate artery (n=2). RESULTS: No stroke occurred. The rate of perioperative myocardial infarction (CKMB rt; 50 U/l) was 5/22. Median ICU length of stay was 54 (15-1245) h. Hospital mortality was 2/22. Pre- and postoperative angina class (CCSC) were 3.3 +/- 0.9 and 1.4 +/- 0.9 respectively (p<0.001). After a median follow up period of 8 months 3 deaths, one stroke, and one myocardial infarction occurred. On 3D multislice CT scan reconstructions which were performed in 13 patients during the first postoperative year all IMA grafts to the LAD and 11 out of 13 extraanatomical vein grafts were shown to be patent. CONCLUSION: Performance of beating heart extraanatomical coronary artery bypass grafts for management of a heavily diseased ascending aorta can result in a very low stroke rate despite a considerable stroke risk. The complexity of the procedures may be reflected by a relatively high rate of perioperative myocardial infarctions. Perioperative mortality as well as short term patency of extraanatomical bypass grafts seem to be acceptable. PMID- 12759203 TI - Integrated Minimally Invasive Direct Coronary Artery Bypass (MIDCAB) grafting and angioplasty for coronary artery revascularization. PMID- 12759204 TI - Conversion from ministernotomy to full sternotomy in aortic valve replacement. AB - Conversion (C) from ministernotomy (M.S.) to full sternotomy was necessary in 5% of the cases in a series of 100 patients consecutively operated for Aortic Valve Replacement (A.V.R.) Analysis of the demographics and surgical techniques indicate older age, aortic fragility, diffuse coronary disease, chronic renal failure and left vent insertion as contributing factors. Despite increased operative blood losses, extra-corporeal circulation (E.C.C.) times, intensive care unit (I.C.U.) stay and hospital stay, no mortality was observed in the conversion group, as compared to 4.2% mortality in the total ministernotomy (MS) population. Preoperative patients selection, avoidance of technical pitfalls, and knowledge of alternative surgical measures are suggested to further decrease the incidence of conversions. PMID- 12759205 TI - Beating heart surgery reduces mortality in the reoperative bypass patient. AB - BACKGROUND: Reoperative coronary artery bypass grafting (redo-CABG) has an increased operative morbidity and mortality compared to patients undergoing primary revascularization. In an effort to reduce the hazards of reoperative CABG, we commenced revascularizing selected patients without cardiopulmonary bypass (CPB) as an alternative to conventional approaches. METHODS: From January 1998 to Dec. 2000, 432 patients underwent reoperative CABG, 153 patients (35%) without the aid of CPB. Treatment groups were compared by means of univariate analysis for preoperative risk factors and postoperative complications. Predicted risk and risk-adjusted mortality were determined by the Society of Thoracic Surgeons risk algorithm. RESULTS: There was a significant difference in the preoperative predicted risk scores between the two treatment groups (off pump 6.5% vs. on pump 5.4%, p=0.0343). There was a significant difference in the off pump observed mortality (2.61%) versus the on pump group (9.68%, p=0.0065). Decreased morbidity in the off pump group was evidenced by a reduced need for blood products (25% vs. 67%, p<0.0001), and the incidence of prolonged ventilation (4% vs. 14%, p=0.0032). The off pump group also had shorter hospital stays (6.2 +/- 5.96 days vs. 8.0 +/- 7.82, p=0.0091). No significant differences between the two groups were seen in the prevalence of perioperative myocardial infarction, stroke, renal failure, or reoperation for bleeding. CONCLUSION: Bypass grafting without CPB significantly decreases mortality and morbidity in selected reoperative patients, and should be considered a viable alternative to conventional approaches. PMID- 12759206 TI - Bypass graft disease: analysis of proliferative activity in human aorto-coronary bypass grafts. AB - BACKGROUND: Aortocoronary bypass graft disease with its increasing clinical signification represents an unsolved problem in cardiological and heart surgery practice. Late occlusion of autologous saphenous vein grafts occurs against a background of medial and neointimal thickening due to migration and proliferation of smooth muscle cells and the later appearance of atherosclerotic plaques. To clarify the role of cellular proliferation in humans we characterized the cellular composition and proliferative index in 30 stenotic saphenous vein grafts. METHODS: 30 stenotic vein grafts and 25 control veins were explantated during redo heart surgery procedures. Time between initial surgical intervention and explantation was 3-168 month (mean 94,8 month). The total area and cell count of the neointima, media and adventitia was calculated computer assisted. Actively proliferating cells were identified using antibody to Ki-67 and by double-lable immuncytochemistry with alpha SMC actin, CD 31 (endothelial cells), CD 68 lpar;makrophages) and CD 45 (T-lymphocytes). RESULTS: Active proliferation was detected in different cell typs with a mean proliferation index of 0.15%,0.18% and 0.086% for the neointima, media and adventita. Only 9% of the proliferating cells in the neointima were SMC (not identified cells 40%); corresponding 14% SMC (not identified cells 33%) were detected in the media. Endothelial cells were the predominante proliferating cell type in all sites of the vessel wall. CONCLUSION: 1. Proliferation occured at low level. While proliferation may play an important role in early lesions our data imply low proliferation activity in advanced graft lesions. Other mechanism like production and deposition of extracellular matrix (ECM) in the neointima are responsible for the lumen reduction of bypass grafts. 2. The high portion of unidentified cells may represent SMC or other cell types at different stages of differentiation; this requires further investigation. 3. The identification of proliferating macrophages and T-lymphocytes implicate an inflammatory component in the development of human bypass graft lesions. PMID- 12759207 TI - Multi-vessel off-pump coronary artery bypass grafting can be taught to trainee surgeons. AB - BACKGROUND: Although, Off-Pump Coronary Artery Bypass (OPCAB) surgery is being increasingly explored and practised in many cardiac units worldwide, there have been only few reports documenting the training of surgeons in this new technique. The purpose of this study was to address the reproducibility of the OPCAB in a unit where this technique is used extensively. METHODS: Registry data, notes and charts of 64 patients who were operated on by four trainee cardiac surgeons over a period of thirteen months at Harefield Hospital, were reviewed retrospectively. These trainees were part of an accredited training programme for cardiothoracic training and were trained by a single consultant trainer in a cardiac unit after it has had an established recent experience in performing non-selective OPCAB for all coming-in patients. Five (7.8%) patients (with 17 distal anastomoses) consented and underwent early postoperative angiography to check the quality of the grafts and anastomoses. RESULTS: The mean age of the study patients was 65.6 and the mean Parsonnet score was 9.4. There was a mean of 2.9 grafts per patient and circumflex territory anastomoses were performed in 48 (75%) patients. No operation required conversion to Cardiopulmonary Bypass (CPB). Angiography of the five patients revealed satisfactory seventeen (100%) distal anastomoses. CONCLUSION: With appropriate training, it is possible for trainees to learn OPCAB and perform multi-vessel revascularization in relatively high-risk patients with good results. PMID- 12759209 TI - A prospective comparison of Doppler echocardiography and postoperative angiography in the assessment of left internal thoracic artery graft in 72 patients submitted to minimally invasive direct coronary artery bypass. AB - BACKGROUND: Concerns have been raised regarding the accuracy of the left internal thoracic artery (LITA) anastomosis performed during minimally invasive direct coronary artery bypass (MIDCAB). In a prospective study, we tested the hypothesis that transthoracic Doppler echocardiography is an adequate technique to determine LITA patency when compared to "gold standard" postoperative angiography. METHODS: Seventy-two consecutive patients with single left anterior descending (LAD) coronary artery stenosis were submitted to MIDCAB performed on a beating heart using the LITA. All patients underwent transthoracic Doppler Echocardiography and angiography before discharge. LITA was considered patent when diastolic fraction (DF) of time-velocity integral was equal or greater than 0.5. FitzGibbon grading system was used to evaluate LITA patency by angiography (A=excellent; B=stenosis reducing caliber of anastomosis or trunk to < 50% of grafted coronary artery; O=occlusion). RESULTS: Angiography showed that LITA was patent in 70 (97.2%) patients, 69 of them been graded A. Adequate image and flow signal of the LITA was achieved in 65 (90.3%) patients, been considered patent in 61 (93.8%) of them. Comparison between echocardiography and angiography in these 65 patients showed a specificity of 96.8% and a sensitivity of 50%. In 7 (9.7%) patients in whom no adequate echocardiography signal was obtained, the LITA graft was normal in six and occluded in one. CONCLUSIONS: For patients whose LITA graft can be imaged, transthoracic Doppler echocardiography is highly specific and is a valuable method for noninvasive evaluation of LITA graft patency after MIDCAB. PMID- 12759208 TI - Continuous graft perfusion: optimizing the quality of saphenous vein grafts. AB - BACKGROUND: Healthy unaltered vascular endothelium in graft material is a prerequisite for a successful CABG operation. Damage done to the endothelium during vein harvest is responsible for an early graft occlusion rate of 20% in the first year after operation. Minimally invasive vein harvesting is regarded to minimize the damage done to the Endothelium. We compared minimally invasive vein harvesting with conventional vein harvesting and studied the influence of a continuous perfusion of the veins with patient autologous blood on their endothelial integrity. METHODS: 80 patients were randomly split into 4 groups: Group 1: Conventional vein harvest and storage of the vein in a crystalloid solution before usage. Group 2: Endoscopic vein harvest and storage in cristallloid solution. Group 3: Conventional harvest under continuous perfusion of the vein with 100 ml blood via the heart lung machine. Group 4: Endoscopic vein harvest under continuous perfusion. Immediately prior to the first peripheral anastomosis a sample was taken from each graft and evaluated by scanning electron microscopy. The endothelial integrity was rated in 5 categories (from "completely confluent endothelium" (1) to "no endothelium" (5)). RESULTS: Group 1: 2.7+/-1.13 Group 2: 2.2+/-1.06 Group 3: 1.6+/-0.68 Group 4: 1.6+/-0.69 CONCLUSION: In regard to the endothelial integrity endoscopic vein harvesting is superior to conventional vein harvest. If the grafts are harvested while continuously perfused with blood there is no more difference between the groups. Considering the well known additional benefits such as reduction in wound healing disorders endoscopic vein harvesting appears to be the preferable technique. PMID- 12759211 TI - Less invasive radial artery harvest. AB - BACKGROUND: Recent studies have sparked a renewed interest in the use of autogenous radial arteries in coronary operations. Some concerns have been found about sequelae of conventional harvesting. A less invasive technique for radial artery harvesting has been proposed by others using endoscopic devices. This technique is time consuming, needs expensive instrumentation and an important learning curve. METHODS: A new less invasive approach for radial harvesting has been developed with a light assisted retractor under direct vision. A small skin incision, median in the forearm, is followed by dissection of the proper plane of the artery. A subcutaneous tunnel is created around the vessel and all the branches are ligated or clipped. The dissection of the pedicle under the skin is completed with the aid of a modified light assisted retractor, originally designed for the saphenous vein harvesting. The incision is closed after heparin reversal with a small redon as drainage. RESULTS: A preliminary serie of 15 patients have been operated with this technique. In all patients the radial artery was patent and functional at the postoperative angiography. Morbidity included only a light hematoma at the beginning of our experience. CONCLUSION: This less invasive technique for the radial artery harvesting appears to be an excellent surgical compromise between the open technique and the endoscopic procedure; it is easy to perform, the learning curve is acceptable and it offers an excellent aesthetic result. PMID- 12759210 TI - Endoscopic vein harvest: early results of a prospective trial with open vein harvest. AB - BACKGROUND: Conventional open long saphenous vein harvest (OVH) is often associated with significant wound pain and serious morbidity in some patients with a resultant extended treatment period. Endoscopic vein harvest (EVH) in theory should alleviate wound pain, be less predisposed towards leg wound infection and lead to greater patient satisfaction. This study aims to compare the two techniques on this basis and determine whether EVH is a viable technique within normal operative time. METHODS: During September and December 2000, 60 saphenous vein harvests were prospectively randomised to EVH (n=30) and OVH (n=30); all performed by one surgeon with the Clearglide(r) endoscopic vein harvest system (Ethicon Inc). End points were impaired wound healing (ASEPSIS score) and postoperative pain (Visual analogue scale) and operative variables. Statistical analysis done using Fisher s exact test and Mann-Whitney U test as appropriate. RESULTS: The groups were well matched demographically. Patients in the EVH group had significantly lower ASEPSIS scores (p<0.001) and postoperative pain (p<0.001). The vein was harvested at 0.96cm/min (0.43-1.5+/-0.33) in the OVH group compared to 0.81cm/min (0.41-1.13+/-0.19) in the EVH group (p=0.09). The new procedure did not prolong the overall operative time (p=0.53). Two patients needed to be converted to open technique. There was no difference found in the vein quality by histological analysis. CONCLUSIONS: These data clearly demonstrate that endoscopic vein harvest results in significantly reduced post operative pain, better impaired wound healing, allows earlier ambulation and does not prolong the operative time significantly with no compromise in vein quality. PMID- 12759212 TI - Echocardiographic findings in minimally invasive coronary artery bypass grafting: The role of intrathoracic CO2 - insufflation and single lung ventilation. AB - BACKGROUND: Current options for surgical treatment of coronary single vessel disease range from beating heart procedure without cardiopulmonary bypass via a mini thoracotomy (MIDCAB) to totally endoscopic robot-assisted techniques (TECAB) with cardiopulmonary bypass. Both procedures are associated with considerable stress even before revascularization such as single lung ventilation, temporary coronary occlusion, Luxatio cordis, intrathoracic CO2 insufflation and extended bypass and operating time. The aim of the this study was to document the extent of intraoperative segmental wall motion abnormalities (SWMA) by echocardiography, and to identify variables affecting SWMA. MATERIALS AND METHODS: Forty patients with coronary single vessel disease were included in the study. 16 patients were operated with the MIDCAB technique, and 24 patients underwent TECAB. In both groups of patients sequential transesophageal echocardiograms (2D-loops) were recorded and analyzed. Hemodynamic and electrocardiographic data as well as oxygenation parameters were acquired during echo exams. In both groups of patients mild, but significant perioperative SWMA were identified, which increased in the course of the operation. These SWMA were more pronounced in the TECAB as compared to the MIDCAB group. Independent of operating time these changes disappeared completely until the ends of surgery. Significant hemodynamic or elektrocardiographic modifications were not observed. CONCLUSION: The application of minimally invasive techniques for the surgical treatment of coronary single vessel disease is associated with significant perioperative SWMA. The more pronounced SWMA in the TECAB group may be a consequence of intrathoracic CO2-insufflation. Both techniques can be applied without significant myocardial ischemia, provided that appropriate intraoperative monitoring is performed, and intrathoracic CO2 pressure in TECAB patients is limited. PMID- 12759213 TI - The left atrial roof incision: an asset for minimally invasive mitral valve surgery. AB - BACKGROUND: The focus in minimally invasive mitral valve surgery has been on reducing thoracic incisions. Several cardiac incisions described in these procedures do not conform with the philosophy of "minimal invasiveness". We describe the left atrial roof incision which has the potential of facilitating an excellent mitral valve exposure through a limited cardiac incision, without major cardiac trauma. The safety, efficacy and technical ease of this technique for minimally invasive mitral valve surgery is evaluated. METHODS: From July 1998 to December 2000, 95 consecutive patients underwent mitral valve surgery by the same surgeon. The patients were divided into 2 groups on the basis of the cardiac incision used for correction of mitral valve disease. Group I patients had left atrial roof incision and group II patients had the traditional paraseptal incision. Each group was further classified into the minimally invasive sternotomy and standard sternotomy subgroups. The clinical variables and complications in the 2 groups were compared. RESULTS: There were 40 male and 55 female patients with a mean age of 63 +/-12yrs (range 29 to 88yrs). LARI was used for mitral valve exposure in 81 patients (85%). In 18 patients (19%), surgery was by minimally invasive technique. The mitral valve was replaced in 82% of patients and repaired in 18%. LARI provided a better exposure of the mitral valve in its anatomical and physiological disposition, without distortion. There was no significant difference in the cross clamp times and postoperative complications between the 2 groups. More patients in the LARI group regained sinus rhythm at discharge. CONCLUSION: LARI is safe and technically easy to perform. It provides an excellent exposure of the mitral valve and left heart cavities in its anatomical disposition with minimal cardiac trauma, making it ideal for minimally invasive mitral surgery. PMID- 12759214 TI - Off-pump redo coronary artery bypass grafting: technical aspects and early results. AB - BACKGROUND: Redo coronary artery bypass grafting (CABG) represents an high-risk surgical procedure, because of an increased incidence of perioperative death, myocardial infarction and stroke. Theoretically, the avoidance of cardiopulmonary bypass may reduce surgical traumatism and ameliorate early results. MATERIALS AND METHODS: From January 1995 to May 2001, we performed 123 redo CABGs, of which 53 (44%) off-pump. Off-pump procedure represented respectively 90% of redo CABG in the period 2000-2001 versus 30% in the 1995-1999 period. The mean age was 66.4 years, males were 39 (73%). The mean 2D-echo ejection fraction was 56% and in 9 cases (17%) was less than 40%. Three operations (5.6%) were performed on an urgent base. The access was median sternotomy in all cases. The mean number of grafts per patient was 1.9 (1.7 in the period 1995-99 vs. 2.3 in the period 2000 01, p=0.01). In 20 cases (38%) we grafted the circumflex artery branches (19% in the period 1995-99 vs. 55.5% in the period 2000-01, p=0.015). Improvements in surgical techniques were achieved over time. The current operative strategy includes the use of deep traction stitches in the posterior pericardium and wall stabilizers to expose target vessels, coronary intraluminal shunts during construction of the anastomoses and continuous trans-esophageal echocardiographic monitoring. Urgent conversion to on-pump procedure was not required in any case. RESULTS: We recorded no in-hospital death, one perioperative myocardial infarction (1.9%), one fifth postoperative day-stroke (1.9%) and 9 atrial fibrillations (17%). Mediastinal re-exploration for bleeding was performed in no one patient; 13 patients (24.5%) required postoperative blood transfusion. The mean length of postoperative stay was 7.5 days, ranging from 6 to 18 days. CONCLUSIONS: In our experience off-pump redo CABG is a safe and effective alternative to on-pump procedure and now off-pump is our first choice-technique in redo CABG. A complete revascularization is technically feasible with a low incidence of perioperative complications. PMID- 12759215 TI - Mitral valve surgery via a right anterior mini-thoracotomy with central aortic cannulation and no endoscopic assistance. AB - BACKGROUND: A variety of techniques have been described to reduce surgical access in mitral valve surgery: Mini-sternotomy (Gundry) involves partial division of the sternum while the right anterior mini-thoracotomy approaches described involve either Port Accesstradmark; (Heartport, Redwood City, CA), indirect endoscopic techniques (Chitwood) and more recently, robotic techniques (Intuitive Surgicaltradmark;). This report describes a simplified approach that "borrows" aspects from several techniques. METHODS: Using currently available technology, a simplified technique to perform mitral valve surgery (MVST) has been developed. MVST eliminates the need for endoscopic assistance and femoral arterial cannulation. The results of 50 of these patients were analyzed and compared with those of 66 patients who had isolated mitral valve surgery via a conventional approach (MVCS) over the same five-year interval. RESULTS: Between January 1, 1995 and December 31, 2000, 50 patients had mitral valve surgery performed with a simplified technique (MVST). Twenty-six (52%) of the MVST patients underwent mitral valve replacement and 24 (48%) underwent mitral valve repair. There were no in-hospital deaths in the MVST group, compared to a death rate of 7.1% in the MVCS group. There were no strokes and no perioperative myocardial infarctions in the MVST group. Average ICU stay was 3.4 days (1 day shorter than the MVCS group) and average hospital stay was 8.1 days, which was significantly less than the 12.5 day length of stay for the patients having MVCS (p<0.01). Blood was utilized in 36% of the MVST patients, compared to a 55% rate in the MVCS group. There were no wound infections in the MVST group. Two patients did develop bloody effusions requiring thoracentesis. Antegrade blood cardioplegia was used in 35 (70%) of the MVST patients. Antegrade and retrograde blood cardioplegia was used in 15 (30%) patients. Average cross-clamp time in the MVST group was 70 minutes compared to 85 in the MVCS group (p<0.05) and the average pump run was 98 minutes in the MVST group compared to 112 for the MVCS group (p=0.08). CONCLUSION: Mitral valve surgery using a simplified, less invasive technique can be successfully and safely performed in selected patients, resulting in less blood utilization and shorter hospital length of stay, with a cosmetic result that rivals that of robotically assisted techniques. PMID- 12759216 TI - Off-pump coronary surgery - results of over 100 cases done by trainees in a single institution over one year. PMID- 12759217 TI - Intraaortic filtration captures particulate debris in OPCAB cases using anastomotic devices. AB - BACKGROUND: Studies associate atheroemboli with neurologic complications following cardiac surgery. The International Council of Emboli Management (ICEM) has demonstrated debris is captured when intraaortic filtration is employed during cardiac surgery. Particulate debris has been extracted from over 98% of ICEM patients and fibrous atheroma from 73%. Anastomotic device use may reduce aortic manipulation, minimizing debris. This study compares particulate capture among three patient groups: Cohort 1 (n = 745) patients receiving on-pump CABG only procedures; Cohort 2 (n = 24) patients receiving off-pump CABG-only procedures with conventional anastomoses; Cohort 3 (n = 10) patients receiving off-pump CABG-only procedures with automatic proximal anastomoses. METHODS: The intraaortic filter was placed distal to anastomoses, and proximal to the innominate artery. Upon removal, filters were fixed in formalin and shipped to a core lab (Stanford, CA). Demographic, procedural, and outcomes data were collected in the ICEM Registry. RESULTS: Of the 745 patients receiving stopped heart CABG, complete data on preoperative risk factors was reported for 673 patients. At least 19 patients reported complete data in Cohort 2, and 10 reported in Cohort 3. One adverse outcome, a death, occurred in the 24 off-pump patients reporting (4.2%). Histologic analyses showing the number of filters successfully extracting particles, the number of particles and total particulate surface area per filter, and the ranges are shown in below. Histologic Finding Cohort 1 Cohort 2 Cohort 3 Filters with particles 98% 100% 100% Mean # of particles 8.5 9.6 5.6 Range # of particles 0-76 1-29 2-13 Mean surface area (mm2) 11.8 18.4 6.1 Range surface area (mm2) 0-171 0.5-61 0.2-14 CONCLUSIONS: These data clearly suggest that there is no difference between the amount of particulate debris generated in OPCAB cases versus CPB cases. In OPCAB cases where an anastomosis device was used without a partial clamp, particulate debris may be reduced but not eliminated. These findings may demonstrate the importance of aortic manipulation, particularly clamping, as a source of particulate material in cardiac surgery. However, more study needs to be done to confirm these findings with a larger number of patients. PMID- 12759218 TI - Angiotensin II phosphorylation of extracellular signal-regulated kinases in rat anterior pituitary cells. AB - We studied the effects of ANG II on extracellular signal-regulated kinase (ERK)1/2 phosphorylation in rat pituitary cells. ANG II increased ERK phosphorylation in a time- and concentration-dependent way. Maximum effect was obtained at 5 min at a concentration of 10-100 nM. The effect of 100 nM ANG II was blocked by the AT1 antagonist DUP-753, by the phospholipase C (PLC) inhibitor U-73122, and by the MAPK kinase (MEK) antagonist PD-98059. The ANG II-induced increase in phosphorylated (p)ERK was insensitive to pertussis toxin blockade and PKC depletion or inhibition. The effect was also abrogated by chelating intracellular calcium with BAPTA-AM or TMB-8 by depleting intracellular calcium stores with a 30-min pretreatment with EGTA and by pretreatment with herbimycin A and PP1, two c-Src tyrosine kinase inhibitors. It was attenuated by AG-1478, an inhibitor of epidermal growth factor receptor (EGFR) activation. Therefore, in the rat pituitary, the increase of pERK is a Gq- and PLC-dependent process, which involves an increase in intracellular calcium and activation of a c-Src tyrosine kinase, transactivation of the EGFR, and the activation of MEK. Finally, the response of ERK activation by ANG II is altered in hyperplastic pituitary cells, in which calcium mobilization evoked by ANG II is also modified. PMID- 12759219 TI - Hormone-sensitive lipase activity and triacylglycerol hydrolysis are decreased in rat soleus muscle by cyclopiazonic acid. AB - Cyclopiazonic acid (CPA) is a sarcoplasmic reticulum Ca2+-ATPase inhibitor that increases intracellular calcium. The role of CPA in regulating the oxidation and esterification of palmitate, the hydrolysis of intramuscular lipids, and the activation of hormone-sensitive lipase (HSL) was examined in isolated rat soleus muscles at rest. CPA (40 micro M) was added to the incubation medium to levels that resulted in subcontraction increases in muscle tension, and lipid metabolism was monitored using the previously described pulse-chase procedure. CPA did not alter the cellular energy state, as reflected by similar muscle contents of ATP, phosphocreatine, free AMP, and free ADP. CPA increased total palmitate uptake into soleus muscle (11%, P < 0.05) and was without effect on palmitate oxidation. This resulted in greater esterification of exogenous palmitate into the triacylglycerol (18%, P < 0.05) and phospholipid (89%, P < 0.05) pools. CPA decreased (P < 0.05) intramuscular lipid hydrolysis, and this occurred as a result of reduced HSL activity (20%, P < 0.05). Incubation of muscles with 3 mM caffeine, which is also known to increase Ca2+ without affecting the cellular energy state, reduced HSL activity (24%, P < 0.05). KN-93, a calcium/calmodulin dependent kinase inhibitor (CaMKII), blocked the effects of CPA and caffeine, and HSL activity returned to preincubation values. The results of the present study demonstrate that CPA simultaneously decreases intramuscular triacylglycerol (IMTG) hydrolysis and promotes lipid storage in isolated, intact soleus muscle. The decreased IMTG hydrolysis is likely mediated by reduced HSL activity, possibly via the CaMKII pathway. These responses are not consistent with the increased hydrolysis and decreased esterification observed in contracting muscle when substrate availability and the hormonal milieu are tightly controlled. It is possible that more powerful signals or a higher [Ca2+] may override the lipid storage effect of the CPA-mediated effects during muscular contractions. PMID- 12759220 TI - TNF-alpha acutely inhibits vascular effects of physiological but not high insulin or contraction. AB - TNF-alpha is elevated in many states of insulin resistance, and acutely administered TNF-alpha in vivo inhibits insulin-mediated hemodynamic effects and glucose uptake in muscle. In this study, we assess whether the inhibitory effects of TNF-alpha are affected by insulin dose or muscle contraction. Whole body glucose infusion rate (GIR), femoral blood flow (FBF), hindleg vascular resistance, hindleg glucose uptake (HGU), 2-deoxyglucose uptake into muscles of the lower leg (R'g) and hindleg metabolism of infused 1-methylxanthine (1-MX), a measure of capillary recruitment, were determined. Three groups were studied with and without infusion of TNF-alpha: euglycemic insulin-clamped, one-leg field stimulated (2 Hz, 0.1 ms at 30 V), and saline-infused control anesthetized rats. Insulin infusions were 3, 10, or 30 mU x kg-1 x min-1 for 2 h x 1-MX metabolism was maximally increased by all three doses of insulin. GIR, HGU, and R'g were maximal at 10 mU and FBF was maximal at 30 mU of insulin. Contraction increased FBF, HGU, and 1-MX. TNF-alpha (0.5 microg x kg-1 x h-1) totally blocked the 3 and 10 mU insulin-mediated increases in FBF and 1-MX, and partly blocked GIR, HGU, and R'g. None of the increases due to twitch contraction was affected by TNF alpha, and only the increase in FBF due to 30 mU of insulin was partly affected. We conclude that muscle capillary recruitment and glucose uptake due to high levels of insulin or muscle contraction under twitch stimuli at 2 Hz are resistant to TNF-alpha. These findings may have implications for ameliorating muscle insulin resistance resulting from increased plasma TNF-alpha and for the differing mechanisms by which contraction and insulin recruit capillary flow in muscle. PMID- 12759221 TI - Combined growth hormone/insulin-like growth factor I in addition to glutamine supplemented TPN results in net protein anabolism in critical illness. AB - Protein loss leading to reduced lean body mass is recognized to contribute to the high levels of morbidity and mortality seen in critical illness. This prospective, randomized, controlled study compared the effects of conventional parenteral nutrition (TPN), glutamine-supplemented (0.4 g.kg-1.day-1) TPN (TPNGLN), and TPNGLN with combined growth hormone (GH, 0.2 IU.kg-1.day-1) and IGF I (160 microg.kg-1.day-1) on protein metabolism in critical illness. Nineteen mechanically ventilated subjects [64 +/- 3 yr, body mass index (BMI) 23.8 +/- 1.3, kg/m2] were initially studied in the fasting state (study 1) and subsequently after 3 days of nutritional with/without hormonal support (study 2). All had recently been admitted to the ICU and the majority were postemergency abdominal surgery (APACHE II 17.5 +/- 1.0). Protein metabolism was assessed using a primed constant infusion of [1-13C]leucine. Conventional TPN contained mixed amino acids, Intralipid, and 50% dextrose. TPNGLN, unlike TPN alone, resulted in an increase in plasma glutamine concentration ( approximately 50%, P < 0.05). Both TPN and TPNGLN decreased the rate of protein breakdown (TPN 15%, P < 0.002; TPNGLN 16%, P < 0.05), but during these treatments the patients remained in a net negative protein balance. Combined treatment with TPNGLN + GH/IGF-I increased plasma IGF-I levels (10.3 +/- 0.8 vs. 48.1 +/- 9.1 nmol/l, study 1 vs. study 2, P < 0.05), and in contrast to therapy with nutrition alone, resulted in net protein gain (-0.75 +/- 0.14 vs. 0.33 +/- 0.12 g protein.kg-1.day-1, study 1 vs. study 2, P < 0.05). Therapy with GH/IGF-I + TPNGLN, unlike nutrition alone, resulted in net positive protein balance in a group of critically ill patients. PMID- 12759222 TI - Differential regional metabolism of glucagon in anesthetized pigs. AB - Glucagon metabolism under basal (endogenous) conditions and during intravenous glucagon infusion was studied in anesthetized pigs by use of midregion (M), COOH terminal (C), and NH2-terminal (N)-RIAs. Arteriovenous concentration differences revealed a negative extraction of endogenous glucagon immunoreactivity across the portal bed (-35.4 +/- 11.0, -40.3 +/- 9.6, -35.6 +/- 16.9%, M-, C-, N-RIA, respectively), reflecting net secretion of pancreatic glucagon and intestinal glicentin and oxyntomodulin, but under exogenous conditions, a net extraction occurred (11.6 +/- 3.6 and 18.6 +/- 5.7%, C- and N-RIA, respectively). Hindlimb extraction of endogenous (17.4 +/- 3.7%, C-RIA) and exogenous (29.1 +/- 4.8 and 19.8 +/- 5.1%, C- and M-RIA) glucagon was detected, indicating M and C cleavage of the molecule. Renal extraction of glucagon was detected by all assays under endogenous (19.4 +/- 6.7, 33.9 +/- 7.1, 29.5 +/- 6.7%, M-, C-, N-RIA) and exogenous conditions (46.9 +/- 4.8, 46.4 +/- 6.0, 47.0 +/- 7.7%; M-, C-, N-RIA), indicating substantial elimination of the peptide. Hepatic glucagon extraction was undetectable under basal conditions and detected only by M-RIA (10.0 +/- 3.8%) during glucagon infusion, indicating limited midregional cleavage of the molecule. The plasma half-life determined by C- and N-RIAs (2.7 +/- 0.2 and 2.3 +/- 0.2 min) were similar, but both were shorter than when determined by M-RIA (3.2 +/- 0.2 min, P < 0.02). Metabolic clearance rates were similar regardless of assay (14.4 +/- 1.1, 13.6 +/- 1.7, 17.0 +/- 1.7 ml x kg-1 x min-1, M-, C-, N RIA). Porcine plasma degraded glucagon, but this was not significantly affected by the dipeptidyl peptidase IV (DPP IV) inhibitor valine-pyrrolidide, and in anesthetized pigs, glucagon's metabolic stability was unchanged by DPP IV inhibition. We conclude that tissue-specific metabolism of glucagon occurs, with the kidney being the main site of removal and the liver playing little, if any, role. Furthermore, valine-pyrrolidide has no effect on glucagon stability, suggesting that DPP IV is unimportant in glucagon metabolism in vivo, in contrast to its significant role in the metabolism of the other proglucagon-derived peptides and glucose-dependent insulinotropic polypeptide. PMID- 12759224 TI - Validation and calibration of DEXA body composition in mice. AB - Validated methods of determining murine body composition are required for studies of obesity in mice. Dual-energy X-ray absorptiometry (DEXA) provides a noninvasive approach to assess body fat and lean tissue contents. Similar to DEXA analyses in other species, body fat measurements in mice show acceptable precision but suffer from poor accuracy. Because fat and lean tissues each contain various components, these inaccuracies likely result from selection of inappropriate calibration standards. Analysis of solvents showed that the PIXImus2 DEXA gave results consistent with theoretical calculations. Male mice weighing 26-60 g and having body fat percentages ranging from 3 to 49% were analyzed by both PIXImus2 DEXA and chemical carcass analysis. DEXA overestimated mouse fat content by an average of 3.3 g, and algorithms were generated to calculate body fat from both measured body fat values and the measured ratio of high- to low-energy X-ray attenuations. With calibration to mouse body fat content measured by carcass analysis, the PIXImus2 DEXA gives accurate body composition values in mice. PMID- 12759223 TI - Physiological role of AMP-activated protein kinase in the heart: graded activation during exercise. AB - AMP-activated protein kinase (AMPK) is emerging as a key signaling pathway that modulates cellular metabolic processes. In skeletal muscle, AMPK is activated during exercise. Increased myocardial substrate metabolism during exercise could be explained by AMPK activation. Although AMPK is known to be activated during myocardial ischemia, it remains uncertain whether AMPK is activated in response to the physiological increases in cardiac work associated with exercise. Therefore, we evaluated cardiac AMPK activity in rats at rest and after 10 min of treadmill running at moderate (15% grade, 16 m/min) or high (15% grade, 32 m/min) intensity. Total AMPK activity in the heart increased in proportion to exercise intensity (P < 0.05). AMPK activity associated with the alpha2-catalytic subunit increased 2.8 +/- 0.4-fold (P < 0.02 vs. rest) and 4.5 +/- 0.6-fold (P < 0.001 vs. rest) with moderate- and high-intensity exercise, respectively. AMPK activity associated with the alpha1-subunit increased to a lesser extent. Phosphorylation of the Thr172-regulatory site on AMPK alpha-catalytic subunits increased during exercise (P < 0.001). There was no increase in Akt phosphorylation during exercise. The changes in AMPK activity during exercise were associated with physiological AMPK effects (GLUT4 translocation to the sarcolemma and ACC phosphorylation). Thus cardiac AMPK activity increases progressively with exercise intensity, supporting the hypothesis that AMPK has a physiological role in the heart. PMID- 12759225 TI - Increased expression of H+-ATPase in inner medullary collecting duct of aquaporin 1-deficient mice. AB - Phenotype analysis has demonstrated that aquaporin-1 (AQP1) null mice are polyuric and manifest a urinary concentrating defect because of an inability to create a hypertonic medullary interstitium. We report here that deletion of AQP1 is also associated with a decrease in urinary pH from 6.15 +/- (SE) 0.1 to 5.63 +/- 0.07. To explore the mechanism of the decrease in urinary pH, we examined the expression of H+-ATPase in kidneys of AQP1 null mice. There was strong labeling for H+-ATPase in intercalated cells and proximal tubule cells in both AQP1 null and wild-type mice. Strong H+-ATPase immunostaining was also present in the apical plasma membrane of inner medullary collecting duct (IMCD) cells in AQP1 null mice, whereas no H+-ATPase labeling was observed in IMCD cells in wild-type mice. In addition, there was an increase in the prevalence of type A intercalated cells in the IMCD of AQP1 null mice, suggesting that the deletion of intercalated cells from the IMCD, which normally occurs during postnatal kidney development, was impaired. Western blot analysis of H+-ATPase expression in the different regions of the kidney demonstrated a significant increase in H+-ATPase protein in the inner medulla of AQP1 null mice compared with wild-type mice. There were no changes in H+-ATPase expression in the cortex or outer medulla. These results represent the first demonstration of apical H+-ATPase immunoreactivity in IMCD cells in vivo and suggest that the decrease in urinary pH observed in AQP1 null mice is due to upregulation of H+-ATPase in the IMCD. The induction of H+-ATPase expression in IMCD cells of AQP1 null mice may be related to the chronically low interstitial osmolality in these animals. The challenge will be to identify the molecular signal(s) responsible for the de novo H+-ATPase expression. PMID- 12759226 TI - Feline interstitial cystitis results in mechanical hypersensitivity and altered ATP release from bladder urothelium. AB - ATP can be released from a variety of cell types by mechanical stimulation; however, the mechanism for this release and the influence of pathology are not well understood. The present study examined intracellular signaling mechanisms involved in swelling-evoked (exposure to a hypotonic solution) release of ATP in urothelial cells from normal cats and cats diagnosed with interstitial cystitis (feline interstitial cystitis; FIC). Using the luciferin-luciferase bioluminescent assay, we demonstrate that swelling-evoked ATP release is significantly elevated in FIC cells. In both normal and FIC cells, ATP release was significantly decreased (mean 70% decrease) by application of blockers of stretch-activated channels (amiloride or gadolinium), as well as brefeldin A and monensin (mean 90% decrease), suggesting that ATP release occurs when ATP containing vesicles fuse with the plasma membrane. Swelling-evoked release was reduced after removal of external calcium (65%), and release was blocked by incubation with BAPTA-AM or agents that interfere with internal calcium stores (caffeine, ryanodine, heparin, or 2-aminoethoxydiphenyl borate). In addition, agents known to act through inositol 1,4,5-triphosphate (IP3) receptors (thapsigargin, acetylcholine) release significantly more ATP in FIC compared with normal urothelium. Taken together, these results suggest that FIC results in a novel hypersensitivity to mechanical stimuli that may involve alterations in IP3 sensitive pathways. PMID- 12759227 TI - Epithelial Na+ channel mutants causing Liddle's syndrome retain ability to respond to aldosterone and vasopressin. AB - Liddle's syndrome is a monogenic form of hypertension caused by mutations in the PY motif of the COOH terminus of beta- and gamma-epithelial Na+ channel (ENaC) subunits. These mutations lead to retention of active channels at the cell surface. Because of the critical role of this PY motif in the stability of ENaCs at the cell surface, we have investigated its contribution to the ENaC response to aldosterone and vasopressin. Mutants of the PY motif in beta- and gamma-ENaC subunits (beta-Y618A, beta-P616L, beta-R564stop, and gamma-K570stop) were stably expressed by retroviral gene transfer in a renal cortical collecting duct cell line (mpkCCDcl4), and transepithelial Na+ transport was assessed by measurements of the benzamil-sensitive short-circuit current (Isc). Cells that express ENaC mutants of the PY motif showed a five- to sixfold higher basal Isc compared with control cells and responded to stimulation by aldosterone (10(-6) M) or vasopressin (10(-9) M) with a further increase in Isc. The rates of the initial increases in Isc after aldosterone or vasopressin stimulation were comparable in cells transduced with wild-type and mutant ENaCs, but reversal of the effects of aldosterone and vasopressin was slower in cells that expressed the ENaC mutants. The conserved sensitivity of ENaC mutants to stimulation by aldosterone and vasopressin together with the prolonged activity at the cell surface likely contribute to the increased Na+ absorption in the distal nephron of patients with Liddle's syndrome. PMID- 12759228 TI - Localization of Mg2+-sensing shark kidney calcium receptor SKCaR in kidney of spiny dogfish, Squalus acanthias. AB - We recently cloned a homologue of the bovine parathyroid calcium receptor from the kidney of a spiny dogfish (Squalus acanthias) and termed this new protein SKCaR. SKCaR senses alterations in extracellular Mg2+ after its expression in human embryonic kidney cells (Nearing J, Betka M, Quinn S, Hentschel H, Elger M, Baum M, Bai M, Chattopadyhay N, Brown E, Hebert S, and Harris HW. Proc Natl Acad. Sci USA 99: 9231-9236, 2002). In this report, we used light and electron microscopic immunocytochemical techniques to study the distribution of SKCaR in dogfish kidney. SKCaR antiserum bound to the apical membranes of shark kidney epithelial cells in the following tubular segments: proximal tubules (PIa and PIIb), late distal tubule, and collecting tubule/collecting duct as well as diffusely labeled cells of early distal tubule. The highly specific distribution of SKCaR in mesial tissue as well as lateral countercurrent bundles of dogfish kidney is compatible with a role for SKCaR to sense local tubular Mg2+ concentrations. This highly specific distribution of SKCaR protein in dogfish kidney could possibly work in concert with the powerful Mg2+ secretory system present in the PIIa segment of elasmobranch fish kidney to affect recycling of Mg2+ from putative Mg2+-sensing/Mg2+-reabsorbing segments. These data provide support for the possible existence of Mg2+ cycling in elasmobranch kidney in a manner analogous to that described for mammals. PMID- 12759229 TI - Smad3 and PKCdelta mediate TGF-beta1-induced collagen I expression in human mesangial cells. AB - Transforming growth factor (TGF)-beta has been associated with fibrogenesis in clinical studies and animal models. We previously showed that Smad3 promotes COL1A2 gene activation by TGF-beta1 in human mesangial cells. In addition to the Smad pathway, it has been suggested that TGF-beta1 could also activate more classical growth factor signaling. Here, we report that protein kinase C (PKC)delta plays a role in TGF-beta1-stimulated collagen I production. In an in vitro kinase assay, TGF-beta1 treatment specifically increased mesangial cell PKCdelta activity in a time-dependent manner. Translocation to the membrane was detected by immunocytochemistry and immunoblot, suggesting activation of PKCdelta by TGF-beta1. Inhibition of PKCdelta by rottlerin decreased basal and TGF-beta1 stimulated collagen I production, mRNA expression, and COL1A2 promoter activity, whereas blockade of conventional PKCs by Go 6976 had little or no effect. In a Gal4-LUC assay system, inhibition of PKCdelta abolished TGF-beta1-induced transcriptional activity of Gal4-Smad3 and Gal4-Smad4(266-552). Overexpression of Smad3 or Smad3D, in which the three COOH-terminal serine phosphoacceptor residues have been mutated, increased activity of the SBE-LUC construct, containing four DNA binding sites for Smad3 and Smad4. This induction was blocked by PKCdelta inhibition, suggesting that rottlerin decreased Smad3 transcriptional activity independently of COOH-terminal serine phosphorylation. Blockade of PKCdelta abolished ligand-independent and ligand-dependent stimulation of COL1A2 promoter activity by Smad3. These data indicate that PKCdelta is activated by TGF-beta1 in human mesangial cells. TGF-beta1-stimulated PKCdelta activity positively regulates Smad transcriptional activity and is required for COL1A2 gene transcription. Thus cross talk among multiple signaling pathways likely contributes to the pathogenesis of glomerular matrix accumulation. PMID- 12759230 TI - Chronic acidosis-induced alteration in bone bicarbonate and phosphate. AB - Chronic metabolic acidosis increases urinary calcium excretion without altering intestinal calcium absorption, suggesting that bone mineral is the source of the additional urinary calcium. In vivo and in vitro studies have shown that metabolic acidosis causes a loss of mineral calcium while buffering the additional hydrogen ions. Previously, we studied changes in femoral, midcortical ion concentrations after 7 days of in vivo metabolic acidosis induced by oral ammonium chloride. We found that, compared with mice drinking only distilled water, ammonium chloride induced a loss of bone sodium and potassium and a depletion of mineral HCO3(-) and phosphate. There is more phosphate than carbonate in neonatal mouse bone. In the present in vitro study, we utilized a high-resolution scanning ion microprobe with secondary ion mass spectroscopy to test the hypothesis that chronic acidosis would decrease bulk (cross-sectional) bone phosphate to a greater extent than HCO3(-) by localizing and comparing changes in bone HCO3(-) and phosphate after chronic incubation of neonatal mouse calvariae in acidic medium. Calvariae were cultured for a total of 51 h in medium acidified by a reduction in HCO3(-) concentration ([HCO(-)]; pH approximately 7.14, [HCO3(-)] approximately 13) or in control medium (pH approximately 7.45, HCO3(-) approximately 26). Compared with incubation in control medium, incubation in acidic medium caused no change in surface total phosphate but a significant fall in cross-sectional phosphate, with respect to the carbon-carbon bond (C2) and the carbon-nitrogen bond (CN). Compared with incubation in control medium, incubation in acidic medium caused no change in surface HCO3(-) but a significant fall in cross-sectional HCO3(-) with respect to C2 and CN. The fall in cross sectional phosphate was significantly greater than the fall in cross-sectional HCO3(-). The fall in phosphate indicates release of mineral phosphates, and the fall in HCO3(-) indicates release of mineral HCO3(-), both of which would be expected to buffer the additional protons and help restore the pH toward normal. Thus a model of chronic acidosis depletes bulk bone proton buffers, with phosphate depletion exceeding that of HCO3(-). PMID- 12759232 TI - Rous-Whipple Award Lecture. How tumors make bad blood vessels and stroma. PMID- 12759233 TI - Intimal exuberance: veins in jeopardy. PMID- 12759234 TI - hTERT gene amplification and increased mRNA expression in central nervous system embryonal tumors. AB - High-level gains at 5p15, a chromosomal region including the human telomerase catalytic protein subunit (hTERT) gene, have been documented in several medulloblastomas. We therefore analyzed hTERT gene dosage in a group of medulloblastomas and other embryonal brain tumors using differential PCR. Amplification of the hTERT locus was detected in 15 of 36 (42%) tumors examined. To correlate gene amplification with message level, we used real-time quantitative PCR to measure hTERT mRNA in 50 embryonal brain tumors. hTERT mRNA was detected in all but one of these cases, and mRNA level correlated significantly with gene dosage (r = 0.82). Log-rank analysis of survival data revealed a trend toward poor clinical outcomes in patients with medulloblastomas containing high hTERT mRNA levels, but clinical follow-up was relatively short and the association was not statistically significant (P = 0.078). Comparative genomic hybridization was used to further analyze the tumor with the greatest hTERT gene dosage and mRNA level, a recurrent medulloepithelioma. hTERT was amplified in the recurrent tumor but not in the primary lesion, suggesting this locus can be involved in tumor progression. Our data indicate that hTERT gene amplification is relatively common in embryonal brain tumors, and that increased expression of hTERT mRNA may be associated with biologically aggressive tumor behavior. PMID- 12759235 TI - Reciprocal modulation of matrix metalloproteinase-13 and type I collagen genes in rat hepatic stellate cells. AB - Collagen degradation by matrix metalloproteinases is the limiting step in reversing liver fibrosis. Although collagen production in cirrhotic livers is increased, the expression and/or activity of matrix metalloproteinases could be normal, increased in early fibrosis, or decreased during advanced liver cirrhosis. Hepatic stellate cells are the main producers of collagens and matrix metalloproteinases in the liver. Therefore, we sought to investigate whether they simultaneously produce alpha1(I) collagen and matrix metalloproteinase-13 mRNAs. In this communication we show that expression of matrix metalloproteinase-13 mRNA is reciprocally modulated by tumor necrosis factor-alpha and transforming growth factor-beta1. When hepatic stellate cells are co-cultured with hepatocytes, matrix metalloproteinase-13 mRNA is up-regulated and alpha1(I) collagen is down regulated. Injuring hepatocytes with galactosamine further increased matrix metalloproteinase-13 mRNA production. Confocal microscopy and differential centrifugation of co-cultured cells revealed that matrix metalloproteinase-13 is localized mainly within hepatic stellate cells. Studies performed with various hepatic stellate cell lines revealed that they are heterogeneous regarding expression of matrix metalloproteinase-13. Those with myofibroblastic phenotypes produce more type I collagen whereas those resembling freshly isolated hepatic stellate cells express matrix metalloproteinase-13. Overall, these findings strongly support the notion that alpha1(I) collagen and matrix metalloproteinase 13 mRNAs are reciprocally modulated. PMID- 12759236 TI - Progesterone induces cellular differentiation in MDA-MB-231 breast cancer cells transfected with progesterone receptor complementary DNA. AB - Progesterone is an important regulator of growth and differentiation in breast tissues. In this study, the effect of progesterone on cell differentiation was evaluated in the estrogen receptor-negative and progesterone receptor (PR) negative MDA-MB-231 cell line which was transfected with PR-complementary DNA. Morphological changes were analyzed at the ultrastructural level by scanning and transmission electron microscopy. Progesterone-treated PR-transfected cells exhibited a more protracted and well spread morphology with an increase in organelles such as mitochondria and rough endoplasmic reticulum as compared to the rounded form of control vehicle (0.1% ethanol)-treated PR-transfected cells. Vehicle and progesterone-treated MDA-MB-231 cells transfected with the pSG5 plasmid (transfection control cells) had similar rounded morphology as control vehicle-treated PR-transfected cells. Immunofluorescence staining revealed that expression of E-cadherin, a differentiation marker, was more prominent in progesterone-treated cells. Expression of keratin and vimentin but not beta catenin was up-regulated in progesterone treated cells when evaluated by immunoblotting. As signal transducers and activators of transcription (STAT) molecules have been implicated in mammary differentiation, we analyzed the expression of Stat 1, 3, 5a, and 5b proteins and found a significant up regulation of the Stat 5b protein in progesterone-treated cells. We have provided in vitro evidence of the close association of PR with differentiation in breast cancer. It is likely that the Stat 5b protein may play a major role in progesterone-induced differentiation in breast cancer cells. PMID- 12759238 TI - Potential tumor suppressive pathway involving DUSP6/MKP-3 in pancreatic cancer. AB - We previously found frequent loss of heterozygosity at 12q21 and 12q22-q23.1 in primary pancreatic cancers, and the DUSP6/MKP-3 gene residing in this region at 12q22 lost its expression in the great majority of pancreatic cancer cell lines. The DUSP6/MKP-3 protein is a dual-specificity phosphatase that dephosphorylates the active form of ERK, making a feedback loop to control ERK activity. Gain-of function mutations of KRAS2 occur in the great majority of pancreatic cancer cells, and loss of expression of DUSP6/MKP-3 may synergistically promote constitutive activation of ERK and uncontrolled cell growth. To study loss of the feedback pathway and its impact on pancreatic cancer cell growth, we first investigated the expression of DUSP6/MKP-3 in primary pancreatic cancer tissues immunohistochemically; we found up-regulation in mildly as well as severely dysplastic/in situ carcinoma cells and down-regulation in invasive carcinoma, especially in the poorly differentiated type. Adenovirus-mediated reintroduction of DUSP6/MKP-3 into cultured pancreatic cancer cells induced strong expression of recombinant DUSP6/MKP-3 and reduction of phosphorylated ERK in a dose-dependent manner based on the multiplicity of infection and resulted in suppression of cell growth. Moreover, analyses by flow cytometry and immunocytochemistry revealed that the exogenous expression of DUSP6/MKP-3 induced apoptosis. These results show that DUSP6 exerts apparent tumor-suppressive effects in vitro and suggest that DUSP6 is a strong candidate tumor suppressor gene at 12q22 locus. PMID- 12759237 TI - Hypoxia-inducible erythropoietin signaling in squamous dysplasia and squamous cell carcinoma of the uterine cervix and its potential role in cervical carcinogenesis and tumor progression. AB - Tissue hypoxia is a characteristic property of cervical cancers that makes tumors resistant to chemo- and radiation therapy. Erythropoietin (Epo) is a hypoxia inducible stimulator of erythropoiesis. Acting via its receptor (EpoR), Epo up regulates bcl-2 and inhibits apoptosis of erythroid cells and rescues neurons from hypoxic damage. In addition to human papillomavirus infection, increased bcl 2 expression and decreased apoptosis are thought to play a role in the progression of cervical neoplasia. Using reverse transcriptase-polymerase chain reaction and Western blotting we showed that HeLa and SiHa cervical carcinoma cells and human cervical carcinomas express EpoR, and that hypoxia enhances EpoR expression. Exogenous Epo stimulated tyrosine phosphorylation and inhibited the cytotoxic effect of cisplatin in HeLa cervical carcinoma cells. Using immunohistochemistry, we examined the expression of Epo, EpoR, p16, hypoxia inducible factor (HIF)-1alpha, and bcl-2 in benign and dysplastic cervical squamous epithelia and invasive squamous cell carcinomas (ISCCs). EpoR expression in benign epithelia was confined to the basal cell layers, whereas in dysplasias it increasingly appeared in more superficial cell layers and showed a significant correlation with severity of dysplasia. Diffuse EpoR expression was found in all ISCCs. Expression of Epo and HIF-1alpha was increased in dysplasias compared to benign epithelia. Focal Epo and HIF-1alpha expression was seen near necrotic areas in ISCCs, and showed correlation in their spatial distribution. Significant correlation was found between expression of EpoR, and p16 and bcl-2 in benign and dysplastic squamous epithelia. Our results suggest that increased expression of Epo and EpoR may play a significant role in cervical carcinogenesis and tumor progression. Hypoxia-inducible Epo signaling may play a significant role in the aggressive behavior and treatment resistance of hypoxic cervical cancers. PMID- 12759239 TI - Protease-activated receptor-2 signaling triggers dendritic cell development. AB - Dendritic cells (DC) are potent antigen-presenting cells that govern the effector cell responses of the immune system. DC are thought to continuously develop from circulating progenitors in a process that is accelerated by inflammatory stimuli. However, the physiological signals that regulate the development of DC from precursor cells have not been well defined. Here we show that a serine protease acting via protease-activated receptor-2 (PAR-2) stimulates the development of DC from bone marrow progenitor cells cultured in granulocyte-macrophage colony stimulating factor and IL-4. DC fail to develop in bone marrow cultures treated with soy bean trypsin inhibitor, a serine protease inhibitor, but this inhibition is overcome by a PAR-2 agonist peptide. DC do not spontaneously develop from the bone marrow of PAR-2-deficient mice, but can be stimulated to do so by inflammatory mediators. These results suggest that endogenous serine proteases stimulate DC development in vitro. Thus, serine proteases may help trigger adaptive immune responses in vivo. PMID- 12759240 TI - Mutation of p53 in recurrent hepatocellular carcinoma and its association with the expression of ZBP-89. AB - p53 has recently been identified as a downstream target of ZBP-89, a zinc finger transcription factor. ZBP-89 promotes growth arrest through stabilization of the p53 protein. The aim of this study is to determine the status of the p53 gene in recurrent human hepatocellular carcinoma (HCC) and test the link between the expression of ZBP-89 and the p53 gene. The results showed that mutations in the p53 gene were frequently detected in recurrent HCC. The interval between surgical resection and the recurrence of HCC was significantly longer in patients with the wild-type p53 gene than those with mutations, strongly suggesting a pathological role for the mutant p53 gene in HCC recurrence. Among those positive for the p53 protein, nearly 85% (18 of 21) showed nuclear localization of the p53 protein while only about 14% (3 of 21) were positive for the p53 protein in the cytoplasm. ZBP-89 co-localized with p53 in the nucleus in about 67% (12 of 18) of all cases positive for the nuclear p53 protein, suggesting that ZBP-89 may play a role in the nuclear accumulation of the p53 protein in a subset of recurrent HCC. With accumulation of p53 protein in the nucleus, tumor cells undergo apoptosis and thus are more susceptible to radiotherapy and chemotherapy. Therefore, co localization of p53 protein with ZBP-89 may define a subgroup of recurrent HCC that is more sensitive to treatment. PMID- 12759241 TI - Matrilysin (matrix metalloproteinase-7) mediates E-cadherin ectodomain shedding in injured lung epithelium. AB - Matrilysin (matrix metalloproteinase-7) is highly expressed in lungs of patients with pulmonary fibrosis and other conditions associated with airway and alveolar injury. Although matrilysin is required for closure of epithelial wounds ex vivo, the mechanism of its action in repair is unknown. We demonstrate that matrilysin mediates shedding of E-cadherin ectodomain from injured lung epithelium both in vitro and in vivo. In alveolar-like epithelial cells, transfection of activated matrilysin resulted in shedding of E-cadherin and accelerated cell migration. In vivo, matrilysin co-localized with E-cadherin at the basolateral surfaces of migrating tracheal epithelium, and the reorganization of cell-cell junctions seen in wild-type injured tissue was absent in matrilysin-null samples. E-cadherin ectodomain was shed into the bronchoalveolar lavage fluid of bleomycin-injured wild-type mice, but was not shed in matrilysin-null mice. These findings identify E-cadherin as a novel substrate for matrilysin and indicate that shedding of E cadherin ectodomain is required for epithelial repair. PMID- 12759242 TI - Constitutive activation of the signal transducer and activator of transcription pathway in celiac disease lesions. AB - The biological effects of interferon (IFN)-gamma rely mainly on the activity of the transcription factor signal transducer and activator of transcription (STAT) 1 and the intracellular levels of suppressor of cytokine signaling (SOCS)-1, a negative regulator that controls the amplitude and duration of STAT-1 activation. IFN-gamma is a key mediator of the immunopathology in celiac disease (CD, gluten sensitive enteropathy). Thus we have investigated STAT-1 signaling and SOCS-1 expression in this condition. As expected, high local concentrations of IFN-gamma were invariably seen in duodenal biopsies from CD patients in comparison to controls. On the basis of immunohistochemistry, STAT-1 phosphorylation, nuclear localization, and DNA-binding activity, STAT-1 activation was consistently more pronounced in CD compared with controls. Despite samples from CD patients containing abundant SOCS-1 mRNA, SOCS-1 protein was expressed at the same level in CD patients and controls. In explant cultures of CD biopsies, gliadin induced the activation of STAT-1 but not SOCS-1. Furthermore, inhibition of STAT-1 prevented the gliadin-mediated induction of ICAM-1 and B7-2. These data suggest that persistent STAT-1 activation can contribute to maintaining and expanding the local inflammatory response in CD. PMID- 12759243 TI - Involvement of large tenascin-C splice variants in breast cancer progression. AB - Alternative splicing of fibronectin-like type III (FNIII) repeats of tenascin-C (Tn-C) generates a number of splice variants. The distribution of large variants, typical components of provisional extracellular matrices that are up-regulated during tumor stroma remodeling, was here studied by immunoblotting and immunohistochemistry using a monoclonal antibody against the FNIII B domain (named 4C8MS) in a series of human breast cancers. Large Tn-C variants were found at only low levels in normal breast tissues, but were highly expressed at invading sites of intraductal cancers and in the stroma of invasive ductal cancers, especially at invasion fronts. There was a positive correlation between the expression of large Tn-C variants and the cell proliferation rate determined by immunolabeling of the Ki-67 antigen. Of the Tn-C recombinant fragments (all FNIII repeats or mFNIII FL, the conserved FNIII domain only, the epidermal growth factor-like domain, and the fibrinogen-like domain) which were expressed by CHO K1 cells transfected with mouse Tn-C cDNAs, only the mFNIII FL enhanced in vitro migration and mitotic activity of mammary cancer cells derived from a Tn-C-null mouse. Addition of 4C8MS blocked the function of mFNIII FL. These findings provide strong evidence that the FNIII alternatively spliced region has important roles in tumor progression of breast cancer. PMID- 12759244 TI - Clearance of apoptotic photoreceptors: elimination of apoptotic debris into the subretinal space and macrophage-mediated phagocytosis via phosphatidylserine receptor and integrin alphavbeta3. AB - The effective phagocytotic clearance of apoptotic debris is fundamental to the maintenance of neural tissues during apoptosis. Retinal photoreceptors undergo apoptosis after retinal detachment. Although their induction phase of apoptosis has been well discussed, their phagocytotic process remains quite unclear. We herein demonstrate that apoptotic photoreceptors are selectively eliminated from their physiological localization, the outer nuclear layer, to the subretinal space, and then phagocytosed by monocyte-derived macrophages. This could be shown by an ultrastructural and immunophenotypic analysis. Moreover, in chimera mice expressing transgenic green fluorescent protein in bone marrow-derived cells, the local infiltration of macrophages could be detected after retinal detachment induced photoreceptor apoptosis. The local injection of an antibody blocking the phosphatidylserine receptor (PSR) or a peptide (GRGDSP)-blocking integrin alphavbeta3 revealed that phagocytotic clearance involves the PSR as well as integrin alphavbeta3 in vivo. Importantly, the level of blockade obtained with these reagents was different. Although anti-PSR increased the frequency of apoptotic cells that fail to bind to macrophages, GRGDSP prevented the engulfment (but not the recognition) of apoptotic photoreceptor cells by macrophages. To our knowledge, this is the first report describing the mechanisms through which apoptotic photoreceptors are selectively eliminated via a directional process in the subretinal space. PMID- 12759245 TI - Differential expression of the angiogenic factor genes vascular endothelial growth factor (VEGF) and endocrine gland-derived VEGF in normal and polycystic human ovaries. AB - Angiogenesis is a key aspect of the dynamic changes occurring during the normal ovarian cycle. Hyperplasia and hypervascularity of the ovarian theca interna and stroma are also prominent features of the polycystic ovary syndrome (PCOS), a leading cause of infertility. Compelling evidence indicated that vascular endothelial growth factor (VEGF) is a key mediator of the cyclical corpus luteum angiogenesis. However, the nature of the factor(s) that mediate angiogenesis in PCOS is less clearly understood. Endocrine gland-derived (EG)-VEGF has been recently identified as an endothelial cell mitogen with selectivity for the endothelium of steroidogenic glands and is expressed in normal human ovaries. In the present study, we compared the expression of EG-VEGF and VEGF mRNA in a series of 13 human PCOS and 13 normal ovary specimens by in situ hybridization. EG-VEGF expression in normal ovaries is dynamic and generally complementary to VEGF expression in both follicles and corpora lutea. A particularly high expression of EG-VEGF was detected in the Leydig-like hilus cells found in the highly vascularized ovarian hilus. In PCOS ovaries, we found strong expression of EG-VEGF mRNA in theca interna and stroma in most of the specimens examined, thus spatially related to the new blood vessels. In contrast, VEGF mRNA expression was most consistently associated with the granulosa cell layer and sometimes the theca, but rarely with the stroma. These findings indicate that both EG-VEGF and VEGF are expressed in PCOS ovaries, but in different cell types at different stages of differentiation, thus suggesting complementary functions for the two factors in angiogenesis and possibly cyst formation. PMID- 12759247 TI - Foveolar differentiation of mouse gastric mucosa in vitro. AB - We report a novel method that allows the culture of highly differentiated gastric surface mucous cells. Isolated mouse gastric epithelial cells and fibroblasts were co-cultured in a three-dimensional collagen gel system, and the reconstructed mucosal surface treated with an air-liquid interface. Cultured cells were examined by histology, immunohistochemistry, and electron microscopy. Isolated epithelial cells were positive for MUC5AC, and showed immature mucous cell features (pre-pit cell stage) on cell-free collagen gel. However, when given fibroblastic support, the epithelial cells differentiated into mature surface mucous cells (pit cell stage), and showed a tall columnar cell shape, basal round nuclei, and mucus-filled cytoplasm. In the fine structure, the cells showed junctional complexes, basal lamina, and glycogen and secretary granules. Further treatment by the air-liquid interface environment modified the differentiated state of the pit cells (pit top cell stage); resulting in the expression of cathepsin E, the disappearance of glycogen granules and the apical accumulation of secretory granules along with an increase in apoptotic cells. This culture model should provide a useful tool for studying gastric epithelial cell biology and various diseases of the gastric mucosa. PMID- 12759246 TI - Heparin inhibits the motility and proliferation of human myometrial and leiomyoma smooth muscle cells. AB - Uterine fibroids (leiomyomas) are a major women's health problem. Currently, the standard for treatment remains hysterectomy, because no other treatment modalities can reduce both symptoms and recurrence. As leiomyomas are a hyperproliferation of smooth muscle cells, we sought to understand the regulation of uterine smooth muscle cell mitogenesis by the glycosaminoglycan heparin, which has been extensively studied as an anti-proliferative molecule in vascular smooth muscle cells. Using matched pairs of human myometrial and leiomyoma smooth muscle cells from the same uterus, we demonstrate that the proliferation and motility of both cell types are inhibited by heparin. We report that the decrease in cell number seen in the presence of heparin is not because of cell death. Interestingly, there is significant patient-to-patient variability in the proliferation response but not in the motility response to heparin. Furthermore, nonanticoagulant and anticoagulant heparin were equally effective at inhibiting leiomyoma and myometrial smooth muscle cell proliferation. These results warrant further investigation into the possibility that heparin might be useful in the treatment of uterine fibroids. PMID- 12759248 TI - Distinct role of fibroblast growth factor-2 and vascular endothelial growth factor on tumor growth and angiogenesis. AB - Tumors express more than a single angiogenic growth factor. To investigate the relative impact of fibroblast growth factor-2 (FGF-2) and vascular endothelial growth factor (VEGF) on tumor growth and neovascularization, we generated tumor cell transfectants differing for VEGF and/or FGF-2 expression. Human endometrial adenocarcinoma HEC-1-B-derived Tet-FGF-2 cells that express FGF-2 under the control of the tetracycline-responsive promoter (Tet-off system) were further transfected with a VEGF(121) anti-sense (AS-VEGF) cDNA. Next, Tet-FGF-2 and AS VEGF/Tet-FGF-2 cells were transplanted subcutaneously in nude mice that received tetracycline or not in the drinking water. Simultaneous expression of FGF-2 and VEGF in Tet-FGF-2 cells resulted in fast-growing lesions characterized by high blood vessel density, patency and permeability, and limited necrosis. Blood vessels were highly heterogeneous in size and frequently associated with pericytes. Inhibition of FGF-2 production by tetracycline caused a significant decrease in tumor burden paralleled by a decrease in blood vessel density and size. AS-VEGF expression resulted in a similar reduction in blood vessel density associated with a significant decrease in pericyte organization, vascular patency, and permeability. The consequent decrease in tumor burden was paralleled by increased tumor hypoxia and necrosis. A limited additional inhibitory effect was exerted by simultaneous down-regulation of FGF-2 and VEGF expression. These findings demonstrate that FGF-2 and VEGF stimulate vascularization synergistically but with distinctive effects on vessel functionality and tumor survival. Blockade of either one of the two growth factors results in a decrease in blood vessel density and, consequently, in tumor burden. However, inhibition of the expression of VEGF, but not of FGF-2, affects also vessel maturation and functionality, leading to tumor hypoxia and necrosis. Our experimental model represents an unique tool to investigate anti-neoplastic therapies in different angiogenic environments. PMID- 12759249 TI - Angiogenic actions of angiopoietin-1 require endothelium-derived nitric oxide. AB - Angiopoietin1 (Ang1) is a novel angiogenic factor with important actions on endothelial cell (EC) differentiation and vascular maturation. Ang1 has been shown to prevent EC apoptosis through activation of PI3-kinase/Akt, a pathway that is also known to activate endothelium nitric oxide synthase (eNOS). Therefore, we hypothesized that the angiogenic effects of Ang1 would also be dependent on the PI3-kinase/Akt pathway, possibly mediated by increased eNOS activity and NO release. Treatment of human umbilical vein endothelial cells with recombinant Ang1* (300 ng/ml) for 15 minutes resulted in PI3-kinase-dependent Akt phosphorylation, comparable to that observed with vascular endothelial growth factor (VEGF) (50 ng/ml), and increased NO production in a PI3-kinase/Akt dependent manner. Capillary-like tube formation induced by Ang1* in fibrin matrix at 24 hours (differentiation index, DI: 13.74 +/- 0.76 versus control 1.71 +/- 0.31) was abolished in the presence of the selective PI3-kinase inhibitor, LY294002 (50 micro mol/L) (DI: 0.31 +/- 0.31, P < 0.01) or the NOS inhibitor, L NAME (3 mmol/L) (DI: 4.10 +/- 0.59, P < 0.01). In subcutaneous Matrigel implants in vivo, addition of recombinant Ang1* or wild-type Ang1 from conditioned media of COS-1 cells transfected with a pFLAG Ang1 expression vector, induced significant neovascularization to a degree similar to VEGF. Finally, angiogenesis in vivo in response to both Ang1 and VEGF was significantly reduced in eNOS deficient compared with wild-type mice. In summary, our results demonstrate for the first time that endothelial-derived NO is required for Ang1-induced angiogenesis, and that the PI3-kinase signaling mediates the activation of eNOS and NO release in response to Ang1. PMID- 12759250 TI - Gelatinase A (MMP-2) is necessary and sufficient for renal tubular cell epithelial-mesenchymal transformation. AB - Progressive renal interstitial fibrosis and tubular atrophy represent the final injury pathway for all commonly encountered forms of renal disease that lead to end-stage renal failure. It has been recently recognized that myofibroblastic cells are the major contributors to the deposition of interstitial collagens. While there are several potential cellular sources of myofibroblasts, attention has focused on the transformation of the organized tubular epithelium to the myofibroblastic phenotype, a process potently driven both in vitro and in vivo by transforming growth factor-beta1 (TGF-beta1). Integrity of the underlying basal lamina provides cellular signals that maintain the epithelial phenotype, and disruption by discrete proteases could potentially initiate the transformation process. We demonstrate that TGF-beta1 coordinately stimulates the synthesis of a specific matrix metalloproteinase, gelatinase A (MMP-2), and its activator protease, MT1-MMP (MMP-14), and that active gelatinase A is absolutely required for epithelial-mesenchymal transformation induced by TGF-beta1. In addition, purified active gelatinase A alone is sufficient to induce epithelial-mesenchymal transformation in the absence of exogenous TGF-beta1. Gelatinase A may also mediate epithelial-mesenchymal transformation in a paracrine manner through the proteolytic generation of active TGF-beta1 peptide. MT1-MMP and gelatinase A were co-localized to sites of active epithelial-mesenchymal transformation and basal lamina disruption in the rat remnant kidney model of progressive renal fibrosis. These studies indicate that a discrete matrix metalloproteinase, gelatinase A, is capable of inducing the complex genetic rearrangements that characterize renal tubular epithelial-mesenchymal transformation. PMID- 12759252 TI - Down-regulation of growth arrest DNA damage-inducible gene 45beta expression is associated with human hepatocellular carcinoma. AB - In this study, we describe the growth arrest DNA damage-inducible gene 45beta (GADD45beta), whose expression was significantly down-regulated in the hepatocellular carcinoma (HCC) microarray study and confirmed by Northern blot analysis. The results suggested that expression of GADD45beta was decreased in human liver cancer cell lines HepG2 and Hep3B, but not in normal human embryonic liver cell line CL-48 or normal liver tissue. Histochemistry study and real-time PCR further confirmed that GADD45beta staining in HCC was significantly decreased when compared to surrounding non-neoplastic liver tissue. In further studies of multiple human cancer tissues, GADD45beta strongly stained tissues such as colon cancer, breast cancer, prostate cancer, squamous cell cancer, lymphoma, and leiomyosarcoma, suggesting that the decreased expression of GADD45beta is specific to HCC. Eighty-five cases of primary HCC were further examined by immunohistochemistry and statistical analyses demonstrated that HCC scored lower than matched non-neoplastic liver tissues consistently and significantly. No staining occurred in 12.94% of HCC cases (score = 0, n = 11); 42.35% had weak staining (score = 1, n = 36); 27.06% had moderate staining (score = 2, n = 23); and 17.65% had staining as strong as normal tissue (score = 3, n = 15). Overall, surrounding non-neoplastic liver tissue was highly positive for GADD45beta compared to adjacent neoplastic liver tissues (P < 0.01). We further observed that down-regulation of GADD45beta expression was strongly correlated with differentiation (P < 0.01) and high nuclear grade (P < 0.01). Moreover, we found that expression of GADD45beta was inversely correlated to the presence of mutant p53 in HCC tissue (P < 0.05). Thus, the results of our study suggest that GADD45beta, which is down-regulated in most cases of HCC, remains an ideal candidate for development as a molecular marker in the diagnosis of HCC and as a potential therapeutic target. PMID- 12759251 TI - Tumor lymphangiogenesis: a novel prognostic indicator for cutaneous melanoma metastasis and survival. AB - Malignant melanomas of the skin are distinguished by their propensity for early metastatic spread via lymphatic vessels to regional lymph nodes, and lymph node metastasis is a major determinant for the staging and clinical management of melanoma. However, the importance of tumor-induced lymphangiogenesis for lymphatic melanoma spread has remained unclear. We investigated whether tumor lymphangiogenesis occurs in human malignant melanomas of the skin and whether the extent of tumor lymphangiogenesis may be related to the risk for lymph node metastasis and to patient survival, using double immunostains for the novel lymphatic endothelial marker LYVE-1 and for the panvascular marker CD31. Tumor samples were obtained from clinically and histologically closely matched cases of primary melanomas with early lymph node metastasis (n = 18) and from nonmetastatic melanomas (n = 19). Hot spots of proliferating intratumoral and peritumoral lymphatic vessels were detected in a large number of melanomas. The incidence of intratumoral lymphatics was significantly higher in metastatic melanomas and correlated with poor disease-free survival. Metastatic melanomas had significantly more and larger tumor-associated lymphatic vessels, and a relative lymphatic vessel area of >1.5% was significantly associated with poor disease-free and overall survival. In contrast, no differences in the density of tumor-associated blood vessels were found. Vascular endothelial growth factor and vascular endothelial growth factor-C expression was equally detected in a minority of cases in both groups. Our results reveal tumor lymphangiogenesis as a novel prognostic indicator for the risk of lymph node metastasis in cutaneous melanoma. PMID- 12759253 TI - Adenoviral gene transfer of interleukin-1 in combination with oncostatin M induces significant joint damage in a murine model. AB - Oncostatin M (OSM) is an interleukin (IL)-6 family cytokine that we have previously shown can synergize with a number of proinflammatory cytokines to promote the release of collagen from cartilage in explant culture. However, the effects of this potent cytokine combination in vivo are not known. Using adenoviral gene transfer, we have overexpressed murine IL-1 (AdmIL-1) and murine OSM (AdmOSM) intraarticularly in the knees of C57BL/6 mice. Histological analyses indicated marked synovial hyperplasia and inflammatory cell infiltration for both AdmIL-1 and AdmOSM but not in control joints. This inflammation was even more pronounced for the AdmIL-1+AdmOSM combination with evidence of cartilage and bone destruction. Significant loss of both proteoglycan and collagen was also seen for this combination, and immunohistochemistry revealed an increased expression of matrix metalloproteinases (MMPs) with decreased tissue inhibitor of metalloproteinases (TIMPs) in both articular cartilage and synovium. Similar expression profiles for MMPs/TIMPs were found in IL-1+OSM-stimulated human articular chondrocytes. Taken together, these data confirm that, in vivo, OSM can exacerbate the effects of IL-1 resulting in inflammation and tissue destruction characteristic of that seen in rheumatoid arthritis. We provide further evidence to implicate the up-regulation of MMPs as a key factor in joint pathology. PMID- 12759254 TI - Nitric oxide synthase 3-dependent vascular remodeling and circulatory dysfunction in cirrhosis. AB - Vascular remodeling is an active process that consists in important modifications in the vessel wall. Endothelium-derived nitric oxide (NO) plays a major role in this phenomenon. We assessed wall thickness (WT), total wall area (TWA), lumen diameter, and total nuclei number/cross-section (TN) in cirrhotic rats with ascites and in control rats. A second group of cirrhotic rats received the NO synthesis inhibitor, L-NAME, or vehicle daily for 11 weeks and systemic hemodynamics, arterial compliance, aortic NO synthase 3 (NOS3) protein expression, and vascular morphology were analyzed. Cirrhotic vessels showed a significant reduction in WT, TWA, and TN as compared to control vessels. Long term inhibition of NOS activity in cirrhotic rats resulted in a significant increase in WT, TWA, and TN as compared to cirrhotic rats receiving vehicle. NOS3 protein abundance was higher in aortic vessels of nontreated cirrhotic animals than in controls. This difference was abolished by chronic treatment with L-NAME. NOS inhibition in cirrhotic rats resulted in higher arterial pressure and peripheral resistance and lower arterial compliance than cirrhotic rats receiving vehicle. Therefore, vascular remodeling in cirrhosis with ascites is a generalized process with significant functional consequences that can be negatively modulated by long-term inhibition of NOS activity. PMID- 12759256 TI - Febrile-range hyperthermia augments pulmonary neutrophil recruitment and amplifies pulmonary oxygen toxicity. AB - Febrile-range hyperthermia (FRH) improves survival in experimental infections by accelerating pathogen clearance, but may also increase collateral tissue injury. We hypothesized that FRH would worsen the outcome of inflammation stimulated by a non-replicating agonist and tested this hypothesis in a murine model of pulmonary oxygen toxicity. Using a conscious, temperature-controlled mouse model, we showed that maintaining a core temperature at FRH (39 degrees C to 40 degrees C) rather than at euthermic levels (36.5 degrees C to 37 degrees C) during hyperoxia exposure accelerated lethal pulmonary vascular endothelial injury, reduced the inspired oxygen threshold for lethality, induced expression of granulocyte-colony stimulating factor, and expanded the circulating neutrophil pool. In these same mice, FRH augmented pulmonary expression of the ELR(+) CXC chemokines, KC and LPS induced CXC chemokine, enhanced recruitment of neutrophils, and changed the histological pattern of lung injury to a neutrophilic interstitial pneumonitis. Immunoblockade of CXC receptor-2 abrogated neutrophil recruitment, reduced pulmonary vascular injury, and delayed death. These combined data demonstrate that FRH may enlist distinct mediators and effector cells to profoundly shift the host response to a defined injurious stimulus, in part by augmenting delivery of neutrophils to sites of inflammation, such as may occur in infections. In certain conditions, such as in the hyperoxic lung, this process may be deleterious. PMID- 12759257 TI - A murine model to study leukocyte rolling and intravascular trafficking in lung microvessels. AB - The cascade of leukocyte interactions under conditions of blood flow is well established in the systemic microcirculation, but not in lung microcirculation. We have developed a murine model to study lung microcirculation by transplanting lung tissue into dorsal skin-fold window chambers in nude mice and examining the ability of leukocytes to traffic within revascularized lung microvessels by intravital microscopy. The revascularized lung allograft demonstrated a network of arterioles, capillaries, and postcapillary venules with continuous blood flow. Stimulation of the lung allograft with TNF-alpha induced leukocyte rolling and adhesion in both arterioles and venules. Treatment with function-blocking anti selectin mAb revealed that P- and L-selectin are the predominant rolling receptors in the lung microvessels, with E-selectin strengthening P-selectin dependent interactions. Intravital microscopic studies also demonstrated that during their transit in capillaries, some leukocytes undergo shape change and continue to roll as elongated cells in postcapillary venules. Furthermore, the revascularized microvessels demonstrated the ability to undergo vasoconstriction in response to superfusion with endothelin-1. Overall, these studies demonstrate that the revascularized lung allograft is responsive to various external stimuli such as cytokines and vaso-active mediators and serves as a model to evaluate the interaction of leukocytes with the vascular endothelium in the lung microcirculation under acute as well as chronic experimental conditions. PMID- 12759255 TI - Experimental diabetes causes breakdown of the blood-retina barrier by a mechanism involving tyrosine nitration and increases in expression of vascular endothelial growth factor and urokinase plasminogen activator receptor. AB - The purpose of these experiments was to determine the specific role of reactive oxygen species (ROS) in the blood-retinal barrier (BRB) breakdown that characterizes the early stages of vascular dysfunction in diabetes. Based on our data showing that high glucose increases nitric oxide, superoxide, and nitrotyrosine formation in retinal endothelial cells, we hypothesized that excess formation of ROS causes BRB breakdown in diabetes. Because ROS are known to induce increases in expression of the well-known endothelial mitogen and permeability factor vascular endothelial growth factor (VEGF) we also examined their influence on the expression of VEGF and its downstream target urokinase plasminogen activator receptor (uPAR). After 2 weeks of streptozotocin-induced diabetes, analysis of albumin leakage confirmed a prominent breakdown of the BRB. This permeability defect was correlated with significant increases in the formation of nitric oxide, lipid peroxides, and the peroxynitrite biomarker nitrotyrosine as well as with increases in the expression of VEGF and uPAR. Treatment with a nitric oxide synthase inhibitor (N-omega-nitro-L-arginine methyl ester, 50 mg/kg/day) or peroxynitrite scavenger (uric acid, 160 mg/kg/day) blocked the breakdown in the BRB and prevented the increases in formation of lipid peroxides and tyrosine nitration as well as the increases in expression of VEGF and uPAR. Taken together, these data indicate that early diabetes causes breakdown of the BRB by a mechanism involving the action of reactive nitrogen species in promoting expression of VEGF and uPAR. PMID- 12759258 TI - Absence of caveolin-1 sensitizes mouse skin to carcinogen-induced epidermal hyperplasia and tumor formation. AB - Caveolin-1 is the principal protein component of caveolae membrane domains, which are located at the cell surface in most cell types. Evidence has accumulated suggesting that caveolin-1 may function as a suppressor of cell transformation in cultured cells. The human CAV-1 gene is located at a putative tumor suppressor locus (7q31.1/D7S522) and a known fragile site (FRA7G) that is deleted in a variety of epithelial-derived tumors. Mechanistically, caveolin-1 is known to function as a negative regulator of the Ras-p42/44 MAP kinase cascade and as a transcriptional repressor of cyclin D1, possibly explaining its transformation suppressor activity in cultured cells. However, it remains unknown whether caveolin-1 functions as a tumor suppressor gene in vivo. Here, we examine the tumor suppressor function of caveolin-1 using Cav-1 (-/-) null mice as a model system. Cav-1 null mice and their wild-type counterparts were subjected to carcinogen-induced skin tumorigenesis, using 7,12-dimethylbenzanthracene (DMBA). Mice were monitored weekly for the development of tumors. We demonstrate that Cav 1 null mice are dramatically more susceptible to carcinogen-induced tumorigenesis, as they develop skin tumors at an increased rate. After 16 weeks of DMBA-treatment, Cav-1 null mice showed a 10-fold increase in tumor incidence, a 15-fold increase in tumor number per mouse (multiplicity), and a 35-fold increase in tumor area per mouse, as compared with wild-type littermate mice. Moreover, before the development of tumors, DMBA-treatment induced severe epidermal hyperplasia in Cav-1 null mice. Both the basal cell layer and the suprabasal cell layers were expanded in treated Cav-1 null mice, as evidenced by immunostaining with cell-type specific differentiation markers (keratin-10 and keratin-14). In addition, cyclin D1 and phospho-ERK1/2 levels were up-regulated during epidermal hyperplasia, suggesting a possible mechanism for the increased susceptibility of Cav-1 null mice to tumorigenesis. However, the skin of untreated Cav-1 null mice appeared normal, without any evidence of epidermal hyperplasia, despite the fact that Cav-1 null keratinocytes failed to express caveolin-1 and showed a complete ablation of caveolae formation. Thus, Cav-1 null mice require an appropriate oncogenic stimulus, such as DMBA treatment, to reveal their increased susceptibility toward epidermal hyperplasia and skin tumor formation. Our results provide the first genetic evidence that caveolin-1 indeed functions as a tumor suppressor gene in vivo. PMID- 12759260 TI - Caveolin-1 knockout mice show an impaired angiogenic response to exogenous stimuli. AB - Recent studies have shown that caveolin-1 (Cav-1) plays an important role as a regulator of angiogenesis in vitro. Here, we use Cav-1 knockout (KO) mice as a model system to examine the in vivo relevance of these findings. A primary mediator of angiogenesis is basic fibroblast growth factor (bFGF). Thus, we studied bFGF-induced angiogenesis in Cav-1 KO mice using a reconstituted basement membrane system, ie, Matrigel plugs, supplemented with bFGF. In Cav-1 KO mice, implanted Matrigel plugs showed a dramatic reduction in both vessel infiltration and density, as compared with identical plugs implanted in wild-type control mice. We also examined the necessity of Cav-1 to support the angiogenic response of an exogenous tumor by subcutaneously injecting Cav-1 KO mice with the melanoma cell line, B16-F10. We show that tumor weight, volume, and vessel density are all reduced in Cav-1 KO mice, consistent with diminished angiogenesis. Ultrastructural analysis of newly formed capillaries within the exogenous tumors reveals a lack of endothelial caveolae and incomplete capillary formation in Cav 1 KO mice. These results provide novel evidence that Cav-1 and caveolae play an important positive role in the process of pathological angiogenesis in vivo. PMID- 12759259 TI - Induction of pathogenic sets of genes in macrophages and neurons in NeuroAIDS. AB - The etiology of the central nervous system (CNS) alterations after human immunodeficiency virus (HIV) infection, such as dementia and encephalitis, remains unknown. We have used microarray analysis in a monkey model of neuroAIDS to identify 98 genes, many previously unrecognized in lentiviral CNS pathogenesis, whose expression is significantly up-regulated in the frontal lobe of simian immunodeficiency virus-infected brains. Further, through immunohistochemical illumination, distinct classes of genes were found whose protein products localized to infiltrating macrophages, endothelial cells and resident glia, such as CD163, Glut5, and ISG15. In addition we found proteins induced in cortical neurons (ie, cyclin D3, tissue transglutaminase, alpha1 antichymotrypsin, and STAT1), which have not previously been described as participating in simian immunodeficiency virus or HIV-related CNS pathology. This molecular phenotyping in the infected brains revealed pathways promoting entry of macrophages into the brain and their subsequent detrimental effects on neurons. These data support the hypothesis that in HIV-induced CNS disease products of activated macrophages and astrocytes lead to CNS dysfunction by directly damaging neurons, as well as by induction of altered gene and protein expression profiles in neurons themselves which are deleterious to their function. PMID- 12759261 TI - Pathogenesis of mucous cell metaplasia in a murine asthma model. AB - Increased mucus production in asthma is an important cause of airflow obstruction during severe exacerbations. To better understand the changes in airway epithelium that lead to increased mucus production, ovalbumin-sensitized and challenged mice were used. The phenotype of the epithelium was dramatically altered, resulting in increased numbers of mucous cells, predominantly in the proximal airways. However, the total numbers of epithelial cells per unit area of basement membrane did not change. A 75% decrease in Clara cells and a 25% decrease in ciliated cells were completely compensated for by an increase in mucous cells. Consequently, by day 22, 70% of the total epithelial cell population in the proximal airways was mucous cells. Electron microscopy illustrated that Clara cells were undergoing metaplasia to mucous cells. Conversely, epithelial proliferation, detected with 5-chloro-2-deoxyuridine immunohistochemistry, was most marked in the distal airways. Using ethidium homodimer cell labeling to evaluate necrosis and terminal dUTP nick-end labeling immunohistochemistry to evaluate apoptosis, this proliferation was accompanied by negligible cell death. In conclusion, epithelial cell death did not appear to be the stimulus driving epithelial proliferation and the increase in mucous cell numbers was primarily a result of Clara cell metaplasia. PMID- 12759263 TI - Chronic otitis media with effusion sequelae in children treated with tubes. AB - OBJECTIVE: To determine incidence and prevalence of middle ear sequelae and abnormal tympanometry results among children with chronic otitis media with effusion (OME) who received standard treatment with tympanostomy tubes. DESIGN: Prospective cohort study. SETTING: Community clinic and academic medical center. Patients A total of 140 children followed up for 8 years after tube treatment. MAIN OUTCOME MEASURES: Tympanic membrane perforation, atrophy, retraction, hearing loss, myringosclerosis, low static admittance (SA) and broad-peaked tympanogram, high SA and narrow-peaked tympanogram, and negative tympanometric peak pressure. RESULTS: Annual incidence of sequelae was typically greater during 3 to 5 years than 6 to 8 years of follow-up. Greatest increases in incidence during the 5-year follow-up were for atrophy (67%), high SA and narrow-peaked tympanogram (70%), and retraction pocket (47%). Prevalence of these sequelae also increased over time, whereas low SA and broad-peaked tympanogram and negative tympanometric peak pressure decreased during follow-up. Sequela tended to become bilateral over time, and concordance of different sequelae in the same ear was low (kappa, 0.05-0.42). CONCLUSIONS: Annual incidence of sequelae decreased during follow-up. This finding parallels decreasing incidence of OME and tube placement as children mature and demonstrates that sequelae are more likely to develop during active acute and chronic OME. The cumulative effect of incidence resulted in few ears free of sequelae by 8 years of follow-up. Based on this cohort of healthy children with OME, although the risk of sequelae decreased over time, functional and morphologic sequelae were prevalent and may put children at risk for continuing middle ear problems as they grow into adolescence and adulthood. PMID- 12759264 TI - Tracheotomy in pediatric patients: a national perspective. AB - BACKGROUND: During the past 50 years, changes in the epidemiology of infectious diseases and the capabilities of medical technology have altered the indications for, and implications of, tracheotomy in children. Given the complexity of health care that these patients subsequently require, monitoring the performance of this procedure and patient outcomes across the diverse US health care system is warranted. OBJECTIVES: To characterize children who received tracheotomies in 1997 and to determine whether disposition and mortality vary by region or health care system attributes. DESIGN: A nationally representative retrospective cohort drawn from an 80% sample of administrative hospital discharge records from all pediatric admissions in 22 states during 1997. PARTICIPANTS: Patients aged 0 to 18 years who underwent tracheotomy. METHODS: The sampling scheme of the discharge records enabled the calculation of regional and national estimates and of age stratified population-based rates of tracheotomies. Weighted descriptive statistical and Poisson analyses were performed. RESULTS: The 2065 tracheotomy procedures recorded in the Kids' Inpatient Database yielded a national estimate of 4861 tracheotomies performed in 1997. The mean length of hospital stay was 50 days, with a mean total facilities charge exceeding $200,000. The rate of tracheotomy was highest among infants and varied significantly across regions of the United States. Adjusting for other patient and health care system attributes, patients who received their tracheotomy in a children's hospital had half the risk of dying during the admission compared with patients who were cared for in a non-children's hospital. Hospitals that performed more pediatric tracheotomies had significantly lower mortality rates than hospitals with lesser case volume. Among patients who survived to discharge, those cared for in the Northeast were discharged to long-term care facilities at twice the rate of patients in the West. Children cared for in children's hospitals or in teaching hospitals were significantly less likely to be discharged to a long-term care facility. CONCLUSIONS: Pediatric tracheotomy is associated with significant variation in rates and outcomes across the United States and across different hospital types. Further research to clarify the reasons for these associations is warranted. PMID- 12759262 TI - Increased venous proinflammatory gene expression and intimal hyperplasia in an aorto-caval fistula model in the rat. AB - We hypothesized that the venous limb of an arteriovenous (AV) fistula would evince up-regulation of genes relevant to vascular remodeling along with neointimal hyperplasia and relevant histological changes. Using the aorto-caval model of an AV fistula model in the rat, we demonstrate marked up-regulation in such proinflammatory genes as monocyte chemoattractant protein-1, plasminogen activator inhibitor-1, and endothelin-1, 2 weeks after the creation of the fistula. Neointimal hyperplasia occurred in variable degrees by 5 weeks after establishing the fistula, and by 16 weeks, such neointimal hyperplasia was progressive and pronounced; at this time point, abundant extracellular matrix was also observed. Smooth muscle cells were present in the hyperplastic neointima as evidenced by staining for alpha-smooth muscle actin; ultrastructurally, smooth muscle cells with a synthetic as well as a contractile phenotype were readily observed. Accumulation of extracellular matrix in the model at 16 weeks was accompanied by increased expression of transforming growth factor-beta1 mRNA, the latter finding contrasting with the suppression of transforming growth factor beta1 mRNA observed in this model at 2 weeks. In summary, we describe marked up regulation in proinflammatory genes and progressive neointimal formation in the venous vasculature in an AV fistula model in the rat. We suggest that such alteration in gene expression and histological injury, in conjunction with the relative simplicity of this model, offer a new approach in the study of such timely biological and clinically relevant phenomena as differential gene expression in response to hemodynamic forces, processes involved in vascular remodeling, mechanisms of injury in venous bypass grafts, and mechanisms of dysfunction of AV fistulae used in hemodialysis. PMID- 12759265 TI - Temperature-controlled radiofrequency tonsil reduction in children. AB - OBJECTIVE: To evaluate the safety and efficacy of temperature-controlled radiofrequency tonsil reduction in the treatment of children with a sleep-related breathing disorder associated with tonsillar obstructive hypertrophy. DESIGN: Prospective, nonrandomized, case series feasibility study of children meeting the criteria for tonsillectomy or adenotonsillectomy for the treatment of an obstructive sleep-related breathing disorder. SETTING: Community-based hospital. Patients Ten children, aged 4 to 13 years, presenting consecutively to a community-based otolaryngology practice with tonsillar or adenotonsillar obstructive hypertrophy implicated clinically in causing a sleep-related breathing disorder; their parents consenting to temperature-controlled radiofrequency tonsil reduction instead of surgical tonsillectomy. Intervention Temperature-controlled radiofrequency tonsil reduction, along with surgical adenoidectomy, if adenoids were present, under general anesthesia. MAIN OUTCOME MEASURES: Tonsil size reduction, treatment morbidity, and symptom improvement with follow-up to 1 year. Baseline and 3-month posttreatment polysomnographic data were used. RESULTS: There was a reduction in tonsil size at 1 year of 75.0% on average, without evidence of regrowth during the 1-year follow-up. All children were drinking liquids in the recovery room, and most were eating soft diets within 6 hours; 8 of the 10 children were eating a normal diet by day 5. On average, the return to normal activity was 3.9 days, with 2.9 days of parental loss of work time. Quality-of-life variables all improved. Snore indexes decreased by 88.6%. Polysomnography at 3 months revealed an 84.2% reduction in the apnea index and a 52.3% reduction in the apnea/hypopnea index. There were no complications. CONCLUSION: Temperature-controlled radiofrequency tonsil reduction seems to be a safe, effective, and minimally morbid treatment for tonsil hypertrophy in children with obstructive sleep-related breathing disorders. PMID- 12759266 TI - The association of the severity of obstructive sleep apnea with plasma leptin levels. AB - OBJECTIVE: To examine whether circulating leptin levels correlate with the severity of disease in patients with obstructive sleep apnea. DESIGN: Prospective nonrandomized study. SETTING: Referral sleep laboratory for patients with sleep disordered breathing and biochemistry laboratory. Patients Thirty-two subjects (mean +/- SD age, 47 +/- 12 years) who were referred for suspected sleep apnea underwent an overnight sleep study and fasting morning venous blood sampling. Patients were divided into 3 groups with respect to apnea-hypopnea index: (1) severe sleep apnea (n = 8), apnea-hypopnea index greater than 20; (2) mild sleep apnea (n = 12), apnea-hypopnea index between 5 and 20; and (3) nonapneic control (n = 12), apnea-hypopnea index less than 5. RESULTS: Leptin levels (mean +/- SD) were 21.2 +/- 8.6, 16.2 +/- 5.2, and 10.6 +/- 7.5 ng/mL (P =.005) in patients with severe and mild obstructive sleep apnea and nonapneic controls, respectively. Plasma leptin levels correlated positively with the degree of sleep disordered breathing as recorded by the apnea-hypopnea index (r = 0.54, P =.001) and percentage of sleep time spent with oxygen saturation below 90% (r = 0.39, P =.02). CONCLUSIONS: Circulating leptin concentrations in patients with obstructive sleep apnea, independent of body mass index and age, are significantly higher than levels in nonapneic controls and there is a positive relationship between leptin concentrations and the severity of sleep apnea. Hyperleptinemia may be a prognostic marker of obstructive sleep apnea syndrome. PMID- 12759267 TI - Efficacy of the 2-staged procedure in the management of cholesteatoma. AB - OBJECTIVE: To demonstrate the efficacy of intact canal wall procedure coupled with a second-stage exploration for the treatment of cholesteatoma. DESIGN: Retrospective case study of patients with cholesteatomas treated with staged surgical extirpation. A minimum of 6 months' postoperative follow-up time was required for inclusion into the study. SETTING: Tertiary academic referral center. Patients A total of 35 adult and pediatric patients, ranging from 9 to 65 years of age, who underwent 2-stage procedures for removal of cholesteatomas. INTERVENTIONS: Two-stage procedures, separated by 6 months, performed with posterior tympanotomy approaches. MAIN OUTCOME MEASURES: The presence or absence of cholesteatoma on second-stage look and the subsequent surgical treatment for recurrent cholesteatoma. The overall hearing results after the completion of the 2-staged procedure were calculated. RESULTS: Disease was controlled in 26 (74%) of the patients. Residual and/or recurrent cholesteatomas were found in 9 (26%) of the patients during the second-stage operation. Of these patients, 5 (14% of the total group) ultimately required conversion to canal-wall-down procedure. Average hearing gain at the completion of the second-stage procedure was 9 dB. CONCLUSIONS: A planned 2-stage procedure that uses the posterior tympanotomy approach for the control of cholesteatoma is an effective technique. This approach offers significant potential for hearing preservation and restoration. PMID- 12759268 TI - Utility of area curve ratio electrocochleography in early Meniere disease. AB - BACKGROUND: Electrocochleography (ECochG) is useful in supporting the diagnosis of Meniere disease (MD). Possible MD (early disease as defined by the 1995 American Academy of Otolaryngology-Head and Neck Surgery Committee on Hearing and Equilibrium) is a readily treatable form of MD. OBJECTIVES: To identify whether ECochG summating potential/action potential (SP/AP) area curve measures are more sensitive than conventional SP/AP amplitude ratios in detecting possible MD. PATIENTS AND METHODS: A retrospective chart review of a 3-year period (1997-2000) was conducted. All charts of patients diagnosed as having MD who had undergone tympanic ECochG were examined to identify those with possible MD. Exclusion criteria were incomplete workup, ECochG performed using a prior system, cochlear microphonic spike obscuring ratio measurements, and prior otologic surgery. A control group of patients with normal SP/AP ratios and ECochG data were identified. SP/AP amplitude and area curve ratios for both groups were measured. RESULTS: Of 138 patients with MD reviewed, 20 (14%) had possible MD, and 8 passed exclusion criteria. An audiologist blinded to patients' diagnoses performed all measurements. The upper limit of normal for SP/AP amplitude and area curve ratios from the control group of ears (n = 13) (alpha =.05) were similar to previously published results. Of the 8 patients with possible MD, 4 had an abnormal SP/AP amplitude ratio, and 7 had an abnormal SP/AP area curve ratio; the difference between groups was statistically significant (P =.03, chi2). CONCLUSIONS: The SP/AP area curve ratio significantly improves ECochG diagnostic sensitivity in possible MD. This ECochG refinement will allow earlier intervention to preserve inner ear function in MD. PMID- 12759269 TI - Temporal bone and sinonasal inverted papilloma: the same pathological entity? AB - BACKGROUND: Inverted papilloma involving the temporal bone is an extremely rare occurrence. Reports in the literature suggest a higher recurrence rate and association with squamous cell carcinoma in Schneiderian-type papillomas of the middle ear than in sinonasal Schneiderian-type papillomas. OBJECTIVES: To investigate the expression of apoptosis-related proteins, markers of cell proliferation activity, and sex hormone receptors in temporal bone inverted papillomas and to compare this entity with sinonasal papillomas. Design and Subjects We investigated 2 rare cases of inverted papilloma of the temporal bone and a control group of 6 cases of sinonasal inverted papilloma. The expression of p53, Mib-1, p27, and progesterone and estrogen receptors was determined. RESULTS: In the 2 cases of temporal bone inverted papilloma, p53 expression was 43.75% and 4.92%; p27 expression was higher in temporal bone inverted papilloma (82.45% and 70.53%) than in the sinonasal inverted papilloma group. One of our 2 cases of temporal bone Schneiderian-type papilloma was positive for progesterone receptor. CONCLUSIONS: The expression of progesterone receptor in 1 of our 2 cases and in the only other case reported in the literature may imply some degree of hormonal dependence of temporal bone inverted papilloma. Our analysis of the expression of apoptosis-related proteins, markers of cell proliferation activity, and sex hormone receptors does not allow us to demonstrate that temporal bone and sinonasal inverted papilloma are different pathological entities. PMID- 12759270 TI - Fluticasone nasal spray and the combination of loratadine and montelukast in seasonal allergic rhinitis. AB - OBJECTIVE: To compare the effectiveness of an intranasal steroid treatment with that of the combination of a histamine1 receptor antagonist and a leukotriene D receptor antagonist in the treatment of seasonal allergic rhinitis. DESIGN: A 2 week, parallel, randomized, double-blind, double-dummy study with rolling enrollment. SETTING: Tertiary care medical center. Subjects A total of 63 adults with a 2-year history of ragweed sensitivity in the Chicago, Ill, area and a positive skin-prick reaction to ragweed pollen. Intervention Subjects were randomized to receive either 100 micro g of fluticasone propionate aqueous nasal spray in each nostril or 10 mg of loratadine and 10 mg of montelukast sodium by mouth once daily in the evening for 2 weeks. At visits 1 and 2, subjects completed a quality-of-life questionnaire and underwent nasal lavage to determine total eosinophil count and eosinophil cationic protein (ECP) measurements. Daily symptom diaries were kept for 2 weeks. MAIN OUTCOME MEASURES: Questionnaire answers, daily nasal symptom scores, eosinophil counts, and ECP levels. RESULTS: Median total nasal symptom scores were lower in the fluticasone group (4.5 vs 6), but the difference was not statistically significant. The questionnaire answers showed dramatic improvement in overall and individual domains for both groups (P<.01 vs visit 1) with significantly greater reduction in nasal symptoms in the fluticasone group (P<.05). Eosinophil counts and ECP levels were significantly reduced in the fluticasone group. CONCLUSION: Both treatments provided clinically meaningful responses, but the overall results favored fluticasone propionate. PMID- 12759271 TI - Effects of exogenous interferon gamma on patients with treatment-resistant chronic rhinosinusitis and dysregulated interferon gamma production: a pilot study. AB - OBJECTIVE: To evaluate the effects of exogenous interferon gamma treatment in patients with chronic rhinosinusitis and evidence of aberrant production of interferon gamma (IFN-gamma) and its regulatory cytokines. METHODS: Ten patients with treatment-resistant chronic rhinosinusitis (4 males and 6 females) treated with exogenous interferon gamma (50 micro g/m2) were retrospectively evaluated by assessing clinical outcomes compared with clinical and laboratory findings before interferon gamma treatment. RESULTS: Dysregulated IFN-gamma production was suspected to be characterized by (1) decreased interleukin 12 production (n = 1), (2) defects in interleukin 12 receptor signaling (n = 4), (3) intrinsic defects in interleukin 12 (n = 4), and (4) decreased IFN-gamma production. Eight patients had a history of chronic otitis media with positive bacterial cultures of sinus lavage samples. Adverse skin reactions to various antibiotics were reported in 7 patients. Asthma was reported in 4 patients. Along with sinusitis symptoms, these conditions were better controlled in all 9 patients who received exogenous interferon gamma for longer than 3 months. In 1 patient, interferon gamma treatment was discontinued after 3 weeks secondary to "presumed" tremor that was later diagnosed as a tic. Repeated surgical procedures and hospitalizations were reported in 2 patients after interferon gamma treatment secondary to recurrent chronic otitis media/mastoiditis/catheter infection and G-tube leakage. Interferon gamma treatment was discontinued in 1 of these patients because of a concern about neutropenia that occurred after catheter infection. Adverse effects of using exogenous interferon gamma were generally limited to local skin reactions. CONCLUSION: Exogenous interferon gamma may be a therapeutic option in a subset of patients with treatment-resistant chronic rhinosinusitis and evidence of dysregulated IFN-gamma production. PMID- 12759272 TI - Endoscopic photography: digital or 35 mm? AB - OBJECTIVE: To compare off-the-shelf digital imaging equipment with a standard single lens reflex 35-mm endoscopic camera in a busy pediatric ears, nose, and throat setting. DESIGN: Two digital cameras with an endoscope adapter and a step down ring were evaluated to obtain optimal settings for digital endoscopic photography. The equipment was used in various clinical and surgical settings to include otoscopy, sinonasal endoscopy, laryngoscopy, and bronchoscopy. The overall quality, color, brightness, and diagnostic quality of the endoscopic digital photographs were compared with those of the single-lens reflex 35-mm flash-generated photographs by experienced endoscopists. Cost analysis and ease of use were also compared. Subjects Initial digital endoscopic settings were formulated from cadaveric tests. These settings were then studied in multiple patients during endoscopy. RESULTS: Endoscopic digital photography resulted in high-quality images in all settings. Digital images were comparable to 35-mm images. The digital system was easier to use and less expensive than the 35-mm system. CONCLUSIONS: We introduce a simple, inexpensive, and easily available endoscopic digital photography system. Digital photography offers numerous advantages over analog photography in a clinical practice. Digital imaging and archiving is more durable and easier to incorporate into patient records and clinical presentations. As the demand for high-quality digital imaging increases, easy-to-use inexpensive digital endoscopic photography will soon replace 35-mm camera technology. PMID- 12759273 TI - Endonasal endoscopic repair of cerebrospinal fluid leaks of the sphenoid sinus. AB - BACKGROUND: Multiple reports have demonstrated the efficacy of the transnasal endoscopic repair of cerebrospinal fluid (CSF) leaks of the anterior cranial base. The literature, however, lacks a comprehensive clinical study specifically addressing the transnasal endoscopic repair of CSF leaks of the sphenoid sinus. OBJECTIVE: To ascertain the factors that significantly affect the surgical outcome after transnasal endoscopic repair of CSF leaks of the sphenoid sinus. METHODS: We retrospectively reviewed the medical records of all patients who underwent an endoscopic transnasal repair of CSF leaks of the sphenoid sinus at our teaching hospitals. RESULTS: Twenty-four patients with CSF leaks of the sphenoid sinuses that were repaired by the transnasal endoscopic approach were included in our study. Causes of the CSF leaks included trauma, surgery, neoplasms, and idiopathic causes. Obliteration was the most common technique used to repair the CSF fistulas (used in 15 [58%] of 26 procedures). Grafting materials included banked pericardium, mucosa, turbinate bone, and mucoperichondrium placed by underlay or onlay grafting or abdominal fat used to obliterate the sphenoid sinus. Twenty-two patients were successfully treated on the first attempt. A persistent leak in 2 patients with previously unrecognized high-pressure hydrocephalus was repaired during a second endoscopic surgery, quickly followed by ventriculoperitoneal shunting. CONCLUSIONS: Assuming an adequate repair, other factors such as the cause, the size of the defect, the technique and material used to repair the defect, and perioperative management do not affect the surgical outcome significantly. Untreated high-pressure hydrocephalus can lead to a recurrence or persistence of the leaks and should be suspected in patients with posttraumatic, idiopathic, or recurrent CSF leaks. PMID- 12759274 TI - Cervical thoracic duct cysts: a case report and review of the literature. PMID- 12759275 TI - Vasomotor rhinitis is not a wastebasket diagnosis. PMID- 12759276 TI - Vasomotor rhinitis remains a true clinical problem. PMID- 12759277 TI - Vasomotor rhinitis should not be a wastebasket diagnosis. PMID- 12759278 TI - Pathology quiz case: cutaneous angiosarcoma. PMID- 12759279 TI - Radiology quiz case 1: patency of Huschke foramen and fistula formation between the auditory canal and the parotid gland. PMID- 12759280 TI - Radiology quiz case 2: Ehlers-Danlos syndrome (EDS) type IV, vascular form. PMID- 12759281 TI - Fenofibrate and losartan. PMID- 12759282 TI - Chondrocalcinosis, osteophytes and osteoarthritis. PMID- 12759284 TI - An unusual cause of pain in both hips. PMID- 12759285 TI - Case number 26: massive cholesterol crystal deposition: unusual location in rheumatoid arthritis. PMID- 12759283 TI - Therapeutic role of dual inhibitors of 5-LOX and COX, selective and non-selective non-steroidal anti-inflammatory drugs. AB - Dual 5-LOX/COX inhibitors are potential new drugs to treat inflammation. They act by blocking the formation of both prostaglandins and leucotrienes but do not affect lipoxin formation. Such combined inhibition avoids some of the disadvantages of selective COX-2 inhibitors and spares the gatrointestinal mucosa. PMID- 12759286 TI - UK community prevalence of knee chondrocalcinosis: evidence that correlation with osteoarthritis is through a shared association with osteophyte. AB - OBJECTIVES: (1) To estimate the prevalence of chondrocalcinosis (CC) in the community and to characterise its compartmental distribution within the knee. (2) To investigate the associations between CC and individual radiographic features of osteoarthritis (OA) at the tibiofemoral joint (TFJ) and patellofemoral joint (PFJ). METHODS: From three community questionnaire studies investigating the prevalence of knee pain, standing anteroposterior and skyline radiographs were obtained on 1727 subjects (1084 women, 643 men; mean age 63.7; 999 (58%) with knee pain). A single observer recorded the presence and site of CC and graded osteophyte and joint space narrowing (JSN) using a line atlas. "OA" was globally defined as the presence of definite osteophyte and definite JSN. Minimum joint space width (JSW) was measured to 0.1 mm with a metered dial caliper RESULTS: (1) The crude prevalence of CC was 7.0% (95% confidence interval (CI) 5.8 to 8.2). This showed a strong association with age. The age adjusted odds ratio (aOR) for CC in women v men was 0.79 (95% CI 0.52 to 1.12). The age, sex, and knee pain standardised estimate for those aged >40 in Nottingham, UK was 4.5%. Patellofemoral CC was seen in only nine cases, all with tibiofemoral CC. (2) The age-sex aOR for the association between CC and OA was 2.08 at the PFJ (1.38 to 3.12) and 2.00 (1.11 to 3.60) at the TFJ. There was no association between measured JSW and CC at either the PFJ or TFJ. Both total osteophyte score and total number of sites with osteophyte were positively associated with CC; aOR for the upper quartile was 2.40 (1.48 to 3.90) and 1.94 (1.15 to 3.26), respectively. An association between CC and diuretic use was also demonstrated (aOR=2.07, 1.02 to 4.19). CONCLUSIONS: In this large UK community study the age, sex, and knee pain adjusted prevalence of CC was 4.5%. There was a strong age association, but no sex predisposition. Patellofemoral CC was uncommon. An association between OA and CC was confirmed, but this appears to operate through an association with osteophyte rather than JSN. The new association between CC and diuretic use might theoretically be explained by diuretic induced hypomagnesaemia. PMID- 12759287 TI - Radiographic assessment of sacroiliitis by radiologists and rheumatologists: does training improve quality? AB - OBJECTIVE: To assess performance of radiologists and rheumatologists in detecting sacroiliitis METHODS: 100 rheumatologists and 23 radiologists participated. One set of films was used for each assessment, another for training, and the third for confidence judgment. Films of HLA-B27+ patients with AS were used to assess sensitivity. For specificity films of healthy HLA-B27- relatives were included. Plain sacroiliac (SI) films with simultaneously taken computed tomographic scans (CTs) were used for confidence judgment. Three months after reading the training set, sensitivity and specificity assessments were repeated. Next, participants attended a workshop. They also rated 26 SI radiographs and 26 CTs for their trust in each judgment. Three months later final assessments were done. RESULTS: Sensitivity (84.3%/79.8%) and specificity (70.6%/74.7%) for radiologists and rheumatologists were comparable. Rheumatologists showed 6.3% decrease in sensitivity after self education (p=0.001), but 3.0% better specificity (p=0.008). The decrease in sensitivity reversed after the workshop. Difference in sensitivity three months after the workshop and baseline was only 0.5%. Sensitivity <50% occurred in 13% of participants. Only a few participants showed changes of >5% in both sensitivity and specificity. Intraobserver agreement for sacroiliitis grade 1 or 2 ranged from 65% to 100%. Sensitivity for CT (86%) was higher than for plain films (72%) (p<0.001) with the same specificity (84%). Confidence ratings for correctly diagnosing presence (7.7) or absence (8.3) of sacroiliitis were somewhat higher than incorrectly diagnosing the presence (6.6) or absence (7.4) of sacroiliitis (p<0.001). CONCLUSION: Radiologists and rheumatologists show modest sensitivity and specificity for sacroiliitis and sizeable intraobserver variation. Overall, neither individual training nor workshops improved performance. PMID- 12759288 TI - Genetic markers for the efficacy of tumour necrosis factor blocking therapy in rheumatoid arthritis. AB - BACKGROUND: Rheumatoid arthritis (RA) is a genetically complex disease where the response to different treatments varies greatly between different patients. This is the case with the tumour necrosis factor (TNF) blocking agents, where 20-40% of patients have been described as non-responders. No predictive markers exist as yet for the prognosis of response. OBJECTIVE: To analyse whether polymorphisms of several cytokine genes are associated with the responsiveness to TNF blockade with etanercept. METHODS: 123 patients with active RA were treated with etanercept and response rates were determined after three months using American College of Rheumatology (ACR)20 and disease activity score (DAS)28 response criteria. Genotyping was done for TNF (-308 TNFA), interleukin (IL)10 (-1087 IL10), transforming growth factor (TGF)beta1 (codon 25 TGFB1), and IL1 receptor antagonist (intron 2 IL1RN). RESULTS: 24 patients (20%) were defined as non responders owing to their failure to fulfil any of the ACR20 or DAS28 response criteria. None of the recorded alleles was alone significantly associated with responsiveness to treatment. However, a certain combination of alleles (-308 TNF1/TNF1 and -1087 G/G) was associated with good responsiveness to etanercept (p<0.05). In addition, a combination of alleles influencing interleukin 1 receptor antagonist (IL1Ra) and TGFbeta1 production (A2 allele for IL1RN and rare C allele in codon 25 of TGFB1 gene) was associated with non-responsiveness (p<0.05). CONCLUSION: Genetic polymorphisms, which may influence the balance of pro- and anti-inflammatory cytokines of relevance for the course of RA, are associated with clinical responsiveness to etanercept treatment. PMID- 12759289 TI - Long term outcome of catastrophic antiphospholipid syndrome survivors. AB - BACKGROUND: Catastrophic antiphospholipid syndrome (APS) is defined as life threatening multiple organ thromboses developing simultaneously or over a short period. The survival rate of catastrophic APS is about 50%, but the long term outcome of patients who survive is unknown. OBJECTIVE: To determine the long term outcome of patients with catastrophic APS and provide further information on patients who survived. PATIENTS AND METHODS: The clinical characteristics and outcomes of 130 patients with catastrophic APS have been reported previously. Six new cases were recently added to this series. Based on these publications, the authors who reported patients who had survived were contacted. Each author was asked (a) what treatment they gave their patients after the catastrophic APS; (b) if their patients had any further thrombosis. RESULTS: 63/136 (46%) patients died at the initial event. Of the remaining 73 patients, information was available for 58 (79%). Thirty eight (66%) patients did not develop further APS related events during an average follow up of 67.2 months. Eleven (19%) patients developed further APS related events but were still alive. No patients developed further catastrophic APS. Nine (16%) patients died: due to multiple organ failure (three patients); myelofibrosis (one); pneumonia (one); and APS related events (four). CONCLUSION: Sixty six per cent of patients who survive an initial catastrophic APS event remained symptom free with anticoagulation during an average follow up of 67.2 months. Twenty six per cent of the survivors developed further APS related events and the mortality rate of these patients was about 25%. PMID- 12759290 TI - Mycophenolate mofetil prevents a clinical relapse in patients with systemic lupus erythematosus at risk. AB - BACKGROUND: Systemic lupus erythematosus (SLE) is characterised by the presence of antibodies to double stranded DNA (dsDNA), which are involved in the pathogenesis of SLE. Previous studies showed that at least two thirds of patients develop a clinical relapse within six months after a significant rise in the anti dsDNA level, and most relapses were prevented by the administration of corticosteroids at the time of the rise. OBJECTIVE: To determine whether mofetil mycophenolate (MMF) can prevent a clinical relapse without the side effects associated with corticosteroids. METHODS: 36 patients with SLE were examined monthly to determine whether a rise in anti-dsDNA level had occurred. A rise was defined as an increase of 25% of the level of the previous sample of at least 15 IU/ml within a four month period. After a rise patients were treated with MMF 2000 mg daily for six months. Patients were monitored monthly for the occurrence of a clinical relapse and to assess the serological activity and state of activation of CD4+, CD8+, and CD19+ lymphocyte subsets. RESULTS: Anti-dsDNA rose in 10 patients. Treatment with MMF was started in all these patients, and after six months no clinical relapse had occurred. Side effects were minimal. Antibodies to dsDNA decreased during the treatment (p<0.001), associated with a decrease in the state of activation of CD19+ lymphocytes. No changes were found in the state of activation of CD4+ or CD8+ lymphocyte subsets. CONCLUSION: Administration of MMF after a rise in antibodies to dsDNA is well tolerated, decreases anti-dsDNA and B cell activation, and seems to prevent the occurrence of a clinical relapse in patients with SLE. PMID- 12759292 TI - Costs of rheumatoid arthritis in Germany: a micro-costing approach based on healthcare payer's data sources. AB - OBJECTIVE: To develop a systematic set of German cost data in rheumatoid arthritis (RA) based solely on valid healthcare payer's cost data sources. METHODS: Retrospectively one year cost data of 338 patients with RA were generated and analysed. The cost data were derived from a major statutory health insurance plan ("Allgemeine Ortskrankenkasse Niedersachsen") and the regional physicians' association ("Kassenarztliche Vereinigung Niedersachsen"). The recently published matrix of cost domains in RA was applied to structure the analysis. Descriptive statistics were used to analyse the data. RESULTS: The total direct costs for the 338 patients during one year (third quarter 2000 to second quarter 2001) were euro 3815 per patient-year. RA related direct costs were euro 2312 per patient-year. Outpatient costs accounted for 73.7%, inpatient costs for 24.0%, and other disease related costs for 2.3% of RA related direct costs. Outpatients cost drivers were RA related drugs (euro 1019 per patient year), physician visits (euro 323 per patient-year), diagnostic and therapeutic procedures and tests (euro 185 per patient-year), and devices and aids (euro;168 per patient-year). 98 patients were retired prematurely owing to RA related work disability and incurred costs of euro;8358 per retired patient-year. 96 patients were gainfully employed and incurred sick leave costs of euro 2835 per employed patient-year. CONCLUSION: Micro-costing based on healthcare payer's data provides a relatively conservative albeit highly accurate estimate of costs in RA. Both RA related and non-RA related costs must be taken into account. In gainfully employed patients and in patients who receive RA related retirement payments productivity costs exceed direct costs. PMID- 12759291 TI - Value of IgA anticardiolipin and anti-beta2-glycoprotein I antibody testing in patients with pregnancy morbidity. AB - OBJECTIVE: To study the prevalence of IgA antiphospholipid antibodies, particularly anticardiolipin antibodies (aCL) and anti-beta(2)-glycoprotein I (abeta(2)GPI), in a cohort of patients with pregnancy morbidity. PATIENTS AND METHODS: Serum samples from four groups of patients were studied by an in house enzyme linked immunosorbent assay (ELISA). Group I: 28 patients with primary antiphospholipid syndrome (PAPS) (median age 32.5 years, range 25-34). Twelve patients had a history of thrombosis. All were positive for IgG/M aCL or lupus anticoagulant (LA), or both. Group II: 28 patients with unexplained pregnancy morbidity (median age 35 years, range 23-48). Seven had history of thrombosis. Nine patients were positive for IgG/M aCL. None from this group fulfilled Sapporo criteria for APS. Group III: 28 patients with systemic lupus erythematosus (SLE) (median age 34 years, range 25-52). Eleven had a history of thrombosis. Twenty one patients had IgG/M aCL and/or LA, but only 19 fulfilled Sapporo criteria for APS. RESULTS: IgA aCL were found in 12, 6, and 14 patients from the groups with PAPS, unexplained pregnancy morbidity, and SLE, respectively. Most patients had these antibodies together with IgG/IgM aCL. Three patients from the group with unexplained pregnancy morbidity and two with SLE had IgA aCL alone. IgA abeta(2)GPI was present in one patient from each group. All IgA abeta(2)GPI were present together with IgG and/or IgM abeta(2)GPI. CONCLUSIONS: The prevalence of IgA aCL is high in patients with pregnancy morbidity, although IgA aCL are usually present together with IgG and/or IgM aCL. IgA abeta(2)GPI are not useful in identifying additional women with APS and pregnancy morbidity. PMID- 12759293 TI - Auditory P300 event related potentials and serotonin reuptake inhibitor treatment in patients with fibromyalgia. AB - BACKGROUND: The P300 components of auditory event related potentials (ERPs) are objective measures related to information and cognitive processing. OBJECTIVES: To assess P300 ERPs in female patients with fibromyalgia (FM) in comparison with healthy age matched controls. To investigate the relationship between P300 potentials and pain threshold levels of patients, and subsequent effect of sertraline treatment on P300 potentials. METHODS: P300 auditory ERPs were studied in 13 untreated female patients with FM and 10 healthy controls matched for age, sex, and education. Pain pressure thresholds and total myalgic scores (TMS) were assessed with an algometer. Patients were evaluated for clinical measures and P300 potentials (recorded from the vertex) at the first visit, and then in the fourth and eighth weeks of sertraline treatment. RESULTS: Patients with FM had significantly lower P300 amplitudes, but not significantly different P300 latencies, than controls at entry. P300 latencies in patients correlated negatively with TMS (r(s)=-0.79, p<0.01) and P300 amplitudes correlated significantly with TMS (r(s)=0.53, p<0.05). Anxiety and depression scores did not correlate significantly with P300 latencies or amplitudes at the study entry. P300 auditory ERPs had increased amplitudes that had reached nearly the same levels as those of the controls at the eighth week without any significant change in their latencies. CONCLUSION: The results show reduced P300 amplitudes in patients with FM. Further studies assessing the relationship between P300 ERPs and neuropsychiatric tests are required for better clarification of the clinical relevance of P300 potentials in FM. PMID- 12759295 TI - Up regulation of the production of tumour necrosis factor alpha and interferon gamma by T cells in ankylosing spondylitis during treatment with etanercept. AB - BACKGROUND: Treatment of active ankylosing spondylitis (AS) with the recombinant, soluble tumour necrosis factor alpha (TNFalpha) receptor molecule etanercept has been shown to be clinically highly effective. The precise mechanism of action, however, is not known. OBJECTIVE: To assess the change in the cytokine secreting ability of CD4+ and CD8+ T cells and macrophages during etanercept treatment. PATIENTS AND METHODS: Peripheral blood mononuclear cells from 10 patients with AS treated with 25 mg etanercept and 10 patients with AS treated with placebo were investigated during treatment given twice weekly subcutaneously. Production of cytokines by T cells was investigated after in vitro stimulation by flow cytometry. RESULTS: Twelve weeks of etanercept treatment induced a significant increase in the number of interferon gamma (IFNgamma) positive (14.2% (9.6-19.5%) before v 24.4% (13.4-36.4%) after) and TNFalpha positive CD4+ T cells (p=0.008 for both cytokines) and IFNgamma positive (37.5% (19.0-45.4%) before v 52.9% (33.2-60.0%) after) and TNFalpha positive CD8+ T cells (p=0.008 for both cytokines) upon phorbol myristate acetate/ionomycin stimulation, but not in the placebo group. Furthermore, etanercept treatment induced a significant increase in the number of IFNgamma positive CD8+ T cells (p=0.024 at 12 weeks) and a non significant increase of TNFalpha positive CD8+ T cells after in vitro stimulation with the aggrecan derived peptides. CONCLUSIONS: Neutralisation of peripheral TNFalpha does not induce a down regulation of the ability of T cells to produce TNFalpha but rather an up regulation, possibly due to a counterregulatory mechanism. PMID- 12759294 TI - Anti-dsDNA, anti-Sm antibodies, and the lupus anticoagulant: significant factors associated with lupus nephritis. AB - BACKGROUND: Lupus nephritis (LN) is a common manifestation in patients with systemic lupus erythematosus (SLE). Autoantibodies and ethnicity have been associated with LN, but the results are controversial. OBJECTIVE: To study the immunological and demographic factors associated with the development of LN. PATIENTS AND METHODS: A retrospective case-control study of 127 patients with biopsy-proven LN, and 206 randomly selected patients with SLE without nephritis as controls was designed. All patients had attended our lupus unit during the past 12 years. Standard methods were used for laboratory testing. RESULTS: Patients with LN were significantly younger than the controls at the time of SLE diagnosis (mean (SD) 25.6 (8.8) years v 33.7 (12.5) years; p<0.0001). The proportion of patients of black ethnic origin was significantly higher in the group with nephritis (p=0.02). There were no differences in sex distribution or duration of follow up. A higher proportion of anti-dsDNA, anti-RNP, anti-Sm, and lupus anticoagulant (LA) was seen in the group with nephritis (p=0.002; p=0.005; p=0.0001; p=0.01, respectively). In univariate, but not in multivariate, analysis male sex and absence of anti-dsDNA were associated with earlier onset of renal disease (p=0.03; p=0.008). In multivariate analysis the only factors associated with nephritis were younger age at diagnosis of SLE, black race, presence of anti dsDNA, anti-Sm, and LA. No demographic or immunological associations were seen with WHO histological classes. CONCLUSIONS: Young, black patients with anti dsDNA, anti-Sm antibodies, and positive LA, appear to have a higher risk of renal involvement. These patients should be carefully monitored for the development of LN. PMID- 12759296 TI - Sonographic training in rheumatology: a self teaching approach. AB - OBJECTIVE: To evaluate a self teaching approach to be followed by a novice without previous practical experience in musculoskeletal ultrasonography. METHODS: The novice was given short general training (two hours) by an experienced sonographer focusing on the approach to the ultrasound equipment, and asked to obtain the best sonographic images of different anatomical areas as similar as possible to the "gold standard" pictures in the online version of the guidelines for musculoskeletal ultrasonography in rheumatology (free access at http://www.sameint.it/eular/ultrasound). At the end of each scanning session, both novice and tutor scored "blindly" all the images from 0 (the lowest quality) to 10 (the highest quality), with a minimum quality score of 6 considered acceptable for standard clinical use. The tutor then explained how to improve the quality of the pictures. Fourteen consecutive inpatients (seven with rheumatoid arthritis, three with psoriatic arthritis, two with reactive arthritis, and two with osteoarthritis) and five healthy subjects were examined. Ultrasound examinations were performed with a Diasus (Dynamic Imaging Ltd, Livingston, Scotland, UK) using two broadband linear probes of 5-10 and 8-16 MHz frequency. RESULTS: Sonographic training lasted one month and included 30 scanning sessions (24 hours of active scanning). 243 images were taken of the selected anatomical areas. The mean time required to produce each image was 6 minutes (SD 4.2; range 1-30). At the end of the training, the novice scored >/=6 for each standard scan. CONCLUSION: A novice can obtain acceptable sonographic images in 24 non consecutive hours of active scanning after an intensive self teaching programme. PMID- 12759297 TI - In vitro growth rate of fibroblast-like synovial cells is reduced by methotrexate treatment. AB - BACKGROUND: Fibroblast-like synovial cells (FLS) can be cultured and expanded in vitro in monolayer. Little is known about the growth characteristics of FLS derived from different patients. OBJECTIVE: To study FLS cultures, with particular attention to differences in growth rate of FLS from patients with rheumatoid arthritis (RA) and from other arthritic patients. Additionally, to analyse the influence of methotrexate (MTX) treatment, patient age, and disease duration on FLS growth characteristics. MATERIALS AND METHODS: FLS were isolated from needle arthroscopy biopsy specimens. Twenty four patients (11 RA, 8 spondyloarthropathy, 1 osteoarthritis, and 4 undifferentiated arthritis) were studied. FLS population doubling time was determined between passage 2 and passage 5. Differences in population doubling time between RA and non-RA FLS and between FLS from patients receiving MTX and those not receiving this drug were analysed. In addition, possible correlations between FLS population doubling time and patient age or disease duration were examined. RESULTS: In vitro monolayer FLS cultures from needle arthroscopy biopsy specimens showed linear growth characteristics. Cell growth rate was not correlated with type of disease. Cells from patients undergoing treatment with MTX showed a longer population doubling time than FLS from patients not receiving this drug (Mann-Whitney test, p<0.05). No correlation was found with patient age or disease duration. CONCLUSION: The results suggest that FLS growth in monolayer is not dependent on the disease affecting the joint. MTX treatment, however, was more relevant in determining FLS growth rate. PMID- 12759298 TI - Effects of combination treatment using anti-hyperuricaemic agents with fenofibrate and/or losartan on uric acid metabolism. AB - OBJECTIVE: To assess the effect of a combination treatment using anti hyperuricaemic agents with fenofibrate and/or losartan on uric acid metabolism in hypertriglyceridaemic and/or hypertensive patients with gout. METHODS: Twenty seven patients with gout were included in a fenofibrate plus anti-hyperuricaemic agents combination study, and 25 in a losartan plus anti-hyperuricaemic agents combination study. Serum uric acid concentration, uric acid clearance, and 24 hour urinary uric acid excretion were measured before and two months after the addition of fenofibrate (300 mg once daily) or losartan (50 mg once daily) to anti-hyperuricaemic agents. RESULTS: Combination therapy of fenofibrate or losartan with anti-hyperuricaemic agents, which included benzbromarone (50 mg once daily) or allopurinol (200 mg twice a day), significantly reduced serum uric acid concentrations in accordance with increased uric acid excretion. CONCLUSION: A combination of fenofibrate or losartan with anti-hyperuricaemic agents is a good option for the treatment of gout patients with hypertriglyceridaemia and/or hypertension, though the additional hypouricaemic effect may be modest. PMID- 12759299 TI - Epidemiological profile of symptomatic osteoarthritis in older adults: a population based study in Dicomano, Italy. AB - OBJECTIVES: To assess the prevalence of hand, hip, and knee osteoarthritis (OA) in an older Italian community dwelling population and its association with disability. METHOD: A cross sectional survey of the whole community aged 65 years and over, was carried out in Dicomano, a small rural town in Tuscany, Italy. Subjects were screened by geriatricians for major chronic conditions, including hip, knee, and hand OA, using diagnostic algorithms based on the American College of Rheumatology (ACR) clinical criteria. A rheumatologist examined subjects who screened positive. Disability was assessed by a World Health Organisation questionnaire. RESULTS: 697 of 864 eligible subjects (81% of the eligible population) were screened. OA of the knee, hand, or hip was identified by clinical ACR criteria respectively in 159, 139, and 81 subjects, and was confirmed by the rheumatologist in 158/182 (87%), 75/101 (74%), and 63.2% of cases. The estimated prevalence was 29.8%, 14.9%, and 7.7%, respectively. Only hip OA was significantly associated with disability in basic activities of daily living. CONCLUSIONS: About one third of community dwelling older people are affected by symptomatic peripheral OA. Hip OA was strongly associated with disability. PMID- 12759300 TI - The "X-Ray RheumaCoach" software: a novel tool for enhancing the efficacy and accelerating radiological quantification in rheumatoid arthritis. AB - BACKGROUND: Precise diagnosis and follow up treatment of rheumatoid arthritis (RA) requires objective quantification, which is still lacking. For this purpose, radiological analyses are considered to be the most appropriate method. OBJECTIVE: To develop computer assisted quantification software that is particularly applicable to joint scoring in rheumatic disorders. METHODS: 3914 radiographs from hands and feet of 190 patients with RA were collected, expertly examined, analysed, and statistically evaluated. Radiographs were quantified using the conventional Larsen score and the "X-Ray RheumaCoach" (XRRC) software. The XRRC is a Java stand alone application which can support and accelerate, but not fully automate, the scoring procedure in RA. The scorer can apply both the Larsen and the Ratingen-Rau scores. RESULTS: Compared with conventional scoring procedures, the XRRC software accelerated quantification time by approximately 25%. The program, which is now available on the internet free of charge, ran stably and proved to be a consistently valuable tool. CONCLUSIONS: Compared with conventional scoring methods, the XRRC software offers several advantages: (a) structured data analysis and input that minimises variance by standardisation; (b) faster and more precise calculation of sum scores and indices; (c) permanent data storing and fast access to the software's database; (d) the possibility of cross calculation to other scores; (e) "user friendly" technology and a dedicated help program; (f) fast access and data transfer through the internet if desired; and (g) reliable documentation of results in a specially designed printout. PMID- 12759301 TI - A neuroimaging follow up study of a patient with juvenile central nervous system systemic lupus erythematosus. AB - BACKGROUND: The course of central nervous system systemic lupus erythematosus (CNS-SLE) is largely unknown. New imaging techniques are available to assist in monitoring the disease course. OBJECTIVE: To report a case of juvenile CNS-SLE, in which magnetic resonance imaging (MRI) was used to assess disease activity. CASE REPORT: A 10-year-old female patient with SLE presented with convulsions; MRI and computed tomography (CT) of the cerebrum disclosed abnormalities. Despite adequate treatment, two years later she had a generalised convulsion, and MRI showed new lesions. MR spectroscopy (MRS) indicated neuronal loss, inflammation, and metabolically compromised tissue; magnetisation transfer imaging (MTI) showed an increase in whole brain lesion load. After exclusion of a malignancy, CNS-SLE was the most likely diagnosis, and cyclophosphamide pulses were administered. Initially, multiple sclerosis (MS)-like lesions regressed, but despite maximal immunosuppressive drugs, new lesions formed and disappeared. When immunosuppressive drugs had been stopped for six months MRI showed improved lesions and MTI histograms. DISCUSSION: In this case report, the anatomical substrate, metabolic aspect, neuroimaging, and clinical course of MS-like lesions in a child with CNS-SLE are described. The way in which radiological techniques can support clinical decision making in this young patient with progressive CNS SLE is illustrated. PMID- 12759302 TI - Successful treatment of severe rheumatoid vasculitis by infliximab. PMID- 12759303 TI - Low dose methotrexate osteopathy in a patient with polyarticular juvenile idiopathic arthritis. PMID- 12759304 TI - More on anticardiolipin and anti-beta2 glycoprotein I in systemic sclerosis. PMID- 12759305 TI - Will pharmacogenetics allow better prediction of methotrexate toxicity and efficacy in patients with RA? PMID- 12759306 TI - Assessment and management of insomnia. PMID- 12759307 TI - Cause of progeria's premature aging found: expected to provide insight into normal aging process. PMID- 12759308 TI - Peter J. Safar, MD: "father of CPR," innovator, teacher, humanist. PMID- 12759313 TI - From the Centers for Disease Control and Prevention. Colorectal cancer test use among persons aged > or =50 years--United States, 2001. PMID- 12759314 TI - From the Centers for Disease Control and Prevention. Advancing HIV prevention: new strategies for a changing epidemic--United States, 2003. PMID- 12759315 TI - From the Centers for Disease Control and Prevention. Update: severe acute respiratory syndrome--United States, 2003. PMID- 12759316 TI - Estimating deaths due to influenza and respiratory syncytial virus. PMID- 12759317 TI - Estimating deaths due to influenza and respiratory syncytial virus. PMID- 12759318 TI - Estimating deaths due to influenza and respiratory syncytial virus. PMID- 12759319 TI - Effect of industry sponsorship on the results of biomedical research. PMID- 12759320 TI - Economic issues in the residency match. PMID- 12759321 TI - Allopurinol and markers of muscle damage among participants in the Tour de France. PMID- 12759322 TI - Active neck muscle training in the treatment of chronic neck pain in women: a randomized controlled trial. AB - CONTEXT: Active physical training is commonly recommended for patients with chronic neck pain; however, its efficacy has not been demonstrated in randomized studies. OBJECTIVE: To evaluate the efficacy of intensive isometric neck strength training and lighter endurance training of neck muscles on pain and disability in women with chronic, nonspecific neck pain. DESIGN: Examiner-blinded randomized controlled trial conducted between February 2000 and March 2002. SETTING: Participants were recruited from occupational health care systems in southern and eastern Finland. PATIENTS: A total of 180 female office workers between the ages of 25 and 53 years with chronic, nonspecific neck pain. INTERVENTIONS: Patients were randomly assigned to either 2 training groups or to a control group, with 60 patients in each group. The endurance training group performed dynamic neck exercises, which included lifting the head up from the supine and prone positions. The strength training group performed high-intensity isometric neck strengthening and stabilization exercises with an elastic band. Both training groups performed dynamic exercises for the shoulders and upper extremities with dumbbells. All groups were advised to do aerobic and stretching exercises regularly 3 times a week. MAIN OUTCOME MEASURES: Neck pain and disability were assessed by a visual analog scale, the neck and shoulder pain and disability index, and the Vernon neck disability index. Intermediate outcome measures included mood assessed by a short depression inventory and by maximal isometric neck strength and range of motion measures. RESULTS: At the 12-month follow-up visit, both neck pain and disability had decreased in both training groups compared with the control group (P<.001). Maximal isometric neck strength had improved flexion by 110%, rotation by 76%, and extension by 69% in the strength training group. The respective improvements in the endurance training group were 28%, 29%, and 16% and in the control group were 10%, 10%, and 7%. Range of motion had also improved statistically significantly in both training groups compared with the control group in rotation, but only the strength training group had statistically significant improvements in lateral flexion and in flexion and extension. CONCLUSIONS: Both strength and endurance training for 12 months were effective methods for decreasing pain and disability in women with chronic, nonspecific neck pain. Stretching and fitness training are commonly advised for patients with chronic neck pain, but stretching and aerobic exercising alone proved to be a much less effective form of training than strength training. PMID- 12759323 TI - Race, quality of care, and outcomes of elderly patients hospitalized with heart failure. AB - CONTEXT: Black patients hospitalized with heart failure reportedly receive poorer quality of care and have worse outcomes than white patients. Because previous studies have been based on selected patient populations treated more than a decade ago, it is unclear if racial differences in quality of care and outcomes currently exist in the United States. OBJECTIVE: To evaluate differences in quality of care and patient outcomes between black and white Medicare beneficiaries hospitalized with heart failure. DESIGN: Retrospective analysis of medical record data systematically collected for the National Heart Failure Project. SETTING AND PATIENTS: Nationwide US sample of 29 732 fee-for-service Medicare beneficiaries hospitalized with heart failure in 1998 and 1999. MAIN OUTCOME MEASURES: Prescription of angiotensin-converting enzyme (ACE) inhibitors, measurement of left ventricular ejection fraction (LVEF), readmission within 1 year of discharge, and mortality within 30 days and 1 year of admission. RESULTS: Black patients and white patients had similar crude rates of LVEF assessment (67.8% black vs 66.6% white; P =.29). Among patients classified as ideal for ACE inhibitor use, black patients had higher crude rates of ACE inhibitor use than white patients (81.0% vs 73.8% white; P<.001) but had similar rates of ACE inhibitor or angiotensin receptor blocker (ARB) use (85.7% black vs 82.5% white; P =.08). After multivariable adjustment, black patients had comparable rates of LVEF assessment (risk ratio [RR], 0.99; 95% confidence interval [CI], 0.95-1.03). Black patients remained more likely to be prescribed ACE inhibitors (RR, 1.22; 95% CI, 1.14-1.28) than were white patients in an adjusted analysis, but there were no significant racial differences in the prescription of ACE inhibitors or ARBs (black vs white, RR, 1.03; 95% CI, 0.97-1.07). Black patients had higher rates of readmission within 1 year of discharge (68.2% vs 63.0%; P<.001) but had lower crude 30-day (6.3% vs 10.7%; P<.001) and 1-year (31.5% vs 40.1%; P<.001) mortality rates than white patients. After multivariable adjustment, black patients had a slightly higher rate of readmission than white patients (RR, 1.09; 95% CI, 1.06-1.13) but remained at lower risk of 30-day mortality (RR, 0.78; 95% CI, 0.68-0.91) and 1-year mortality (RR, 0.93; 95% CI, 0.88-0.98). CONCLUSIONS: Black Medicare patients hospitalized with heart failure received comparable quality of care and had slightly higher rates of readmission but had lower mortality rates up to 1 year after hospitalization than did white patients. PMID- 12759324 TI - Combination therapy with hormone replacement and alendronate for prevention of bone loss in elderly women: a randomized controlled trial. AB - CONTEXT: Therapy with individual antiresorptive agents has been shown to be effective for prevention and treatment of postmenopausal osteoporosis, but whether combination antiresorptive therapy with hormones and bisphosphonates is safe or efficacious or how these agents compare in elderly women is unknown. OBJECTIVE: To determine whether hormone replacement and the bisphosphonate alendronate sodium in combination are efficacious and safe, and how they compare with monotherapy in community-dwelling elderly women. DESIGN: Randomized, double blind, placebo-controlled, clinical trial. SETTING AND PARTICIPANTS: Five hundred seventy-three community-dwelling women age 65 years or older were screened: 485 completed screening and 373 (aged 65 to 90 years) were randomized following a 3 month, open-label, run-in phase with hormone replacement and alendronate placebo. The trial was conducted at a single academic US medical center from January 1996 to May 2001. INTERVENTIONS: Participants were randomly assigned in a 2 x 2 factorial design to receive hormone replacement (conjugated equine estrogen, 0.625 mg/d, with or without medroxyprogesterone, 2.5 mg/d) and alendronate, 10 mg daily, both agents, or neither. All participants received calcium and vitamin D supplements. MAIN OUTCOME MEASURES: Annualized change in bone mineral density of the hip and spine and occurrence of adverse events. RESULTS: Bone mineral density at 3 years was significantly greater at all femoral and vertebral sites in women treated with combination therapy than with monotherapy, with mean (SD) increases of 5.9% (3.8) at the total hip, 10.4% (5.4) at the posteroanterior lumbar spine, and 11.8% (6.8) at the lateral lumbar spine. Mean (SD) increases in bone mass at the hip in women treated with alendronate alone were significantly greater than in those treated with hormone replacement therapy alone (4.2% [3.8] vs 3.0% [4.9]; P<.05, respectively), and alendronate resulted in more responders to therapy. All therapies were well tolerated and participant retention was 90% at 3 years. CONCLUSIONS: Combination therapy with hormone replacement and alendronate was efficacious and well tolerated in this cohort. Alendronate was superior to hormone replacement, and combination therapy was superior to either therapy alone. Combination therapy may represent an option for women with more severe disease or for those who have failed to achieve an adequate response to monotherapy. PMID- 12759325 TI - Health outcomes associated with various antihypertensive therapies used as first line agents: a network meta-analysis. AB - CONTEXT: Establishing relative benefit or harm from specific antihypertensive agents is limited by the complex array of studies that compare treatments. Network meta-analysis combines direct and indirect evidence to better define risk or benefit. OBJECTIVE: To summarize the available clinical trial evidence concerning the safety and efficacy of various antihypertensive therapies used as first-line agents and evaluated in terms of major cardiovascular disease end points and all-cause mortality. DATA SOURCES AND STUDY SELECTION: We used previous meta-analyses, MEDLINE searches, and journal reviews from January 1995 through December 2002. We identified long-term randomized controlled trials that assessed major cardiovascular disease end points as an outcome. Eligible studies included both those with placebo-treated or untreated controls and those with actively treated controls. DATA EXTRACTION: Network meta-analysis was used to combine direct within-trial between-drug comparisons with indirect evidence from the other trials. The indirect comparisons, which preserve the within-trial randomized findings, were constructed from trials that had one treatment in common. DATA SYNTHESIS: Data were combined from 42 clinical trials that included 192 478 patients randomized to 7 major treatment strategies, including placebo. For all outcomes, low-dose diuretics were superior to placebo: coronary heart disease (CHD; RR, 0.79; 95% confidence interval [CI], 0.69-0.92); congestive heart failure (CHF; RR, 0.51; 95% CI, 0.42-0.62); stroke (RR, 0.71; 0.63-0.81); cardiovascular disease events (RR, 0.76; 95% CI, 0.69-0.83); cardiovascular disease mortality (RR, 0.81; 95% CI, 0.73-0.92); and total mortality (RR, 0.90; 95% CI, 0.84-0.96). None of the first-line treatment strategies-beta-blockers, angiotensin-converting enzyme (ACE) inhibitors, calcium channel blockers (CCBs), alpha-blockers, and angiotensin receptor blockers-was significantly better than low-dose diuretics for any outcome. Compared with CCBs, low-dose diuretics were associated with reduced risks of cardiovascular disease events (RR, 0.94; 95% CI, 0.89-1.00) and CHF (RR, 0.74; 95% CI, 0.67-0.81). Compared with ACE inhibitors, low-dose diuretics were associated with reduced risks of CHF (RR, 0.88; 95% CI, 0.80-0.96), cardiovascular disease events (RR, 0.94; 95% CI, 0.89-1.00), and stroke (RR, 0.86; 0.77-0.97). Compared with beta-blockers, low-dose diuretics were associated with a reduced risk of cardiovascular disease events (RR, 0.89; 95% CI, 0.80-0.98). Compared with alpha-blockers, low-dose diuretics were associated with reduced risks of CHF (RR, 0.51; 95% CI, 0.43-0.60) and cardiovascular disease events (RR, 0.84; 95% CI, 0.75-0.93). Blood pressure changes were similar between comparison treatments. CONCLUSIONS: Low-dose diuretics are the most effective first-line treatment for preventing the occurrence of cardiovascular disease morbidity and mortality. Clinical practice and treatment guidelines should reflect this evidence, and future trials should use low-dose diuretics as the standard for clinically useful comparisons. PMID- 12759326 TI - Analysis and reporting of factorial trials: a systematic review. AB - CONTEXT: Although factorial trials have become common, standards for the analysis and reporting of such trials have not been established and, despite concerns about the possibility of unrecognized interactions between therapies in factorial trials, the magnitude of this potential problem is unknown. OBJECTIVE: To examine the rationale, methods, and analysis of randomized factorial trials. DATA SOURCES AND STUDY SELECTION: We searched MEDLINE, EMBASE, and the Cochrane Controlled Trials Register using the terms factorial, interaction, 2 x 2, 2 by 2, and incremental to identify factorial randomized trials published from January 2000 to July 2002. To identify trials missed by the electronic search, we performed a hand search of English-language trials in a defined topic area (using the term myocardial ischemia [exp]) listed in MEDLINE (1966-2002), EMBASE (1980-2002), and the Cochrane Controlled Trials Register, as well as all trials in any topic area published in December 2000, excluding trials reporting only continuous surrogate end points. The final set of 33 eligible publications described 29 unique trials. DATA EXTRACTION: Two investigators independently identified factorial trials, generated a list of items affecting validity of results, and abstracted these items from each trial. DATA SYNTHESIS: The sensitivity of electronic searching for identifying factorial trials was 76%. Our 3-pronged search strategy identified 44 factorial trials with clinically important binary outcomes: 36 (82%) were done for reasons of efficiency (testing 2 interventions in the same patient population), and 8 (18%) were done to assess the incremental benefits of combining the 2 treatments. All but 1 of the trials reported treatment effects by comparing all patients who received treatment A (ie, those receiving either A alone or both A and B) vs all those not receiving treatment A (ie, those receiving either B alone or neither A nor B). Twenty-nine of the 44 trials (66%) reported the data from each of the treatment groups separately; 26 trials (59%) reported testing for interactions between the treatments. Only 2 of 31 (6%) comparisons demonstrated a statistically significant interaction between the 2 treatments. CONCLUSIONS: Accurate interpretation of factorial trials depends on the transparent reporting of data for each treatment cell. Despite concerns about unrecognized interactions, our findings suggest that investigators are appropriately restricting their use of the factorial design to those situations in which 2 (or more) treatments do not have the potential for substantive interaction. PMID- 12759327 TI - Composite outcomes in randomized trials: greater precision but with greater uncertainty? AB - Composite outcomes, in which multiple end points are combined, are frequently used as primary outcome measures in randomized trials and are often associated with increased statistical efficiency. However, such measures may prove challenging for the interpretation of results. In this article, we examine the use of composite outcomes in major clinical trials, assess the arguments for and against them, and provide guidance on their application and reporting. To assess incidence and quality of reporting, we systematically reviewed the use of composite end points in clinical trials in Annals of Internal Medicine, BMJ, Circulation, Clinical Infectious Diseases, Journal of the American College of Cardiology, JAMA, Lancet, New England Journal of Medicine, and Stroke from 1997 through 2001 using a sensitive search strategy. We selected for review 167 original reports of randomized trials (with a total of 300 276 patients) that included a composite primary outcome that incorporated all-cause mortality. Sixty three trials (38%) were neutral both for the primary end point and the mortality component. Sixty trials (36%) reported significant results for the primary outcome measure but not for the mortality component. Only 6 trials (4%) were significant for the mortality component but not for the primary composite outcome, whereas 19 trials (11%) were significant for both. Twenty-two trials (13%) were inadequately reported. Our review suggests that reporting of composite outcomes is generally inadequate, implying that the results apply to the individual components of the composite outcome rather than only to the overall composite. Current guidelines for the undertaking and reporting of clinical trials could be revised to reflect the common use of composite outcomes in clinical trials. PMID- 12759328 TI - Clinical trials--multiple treatments, multiple end points, and multiple lessons. PMID- 12759329 TI - JAMA patient page. Insomnia. PMID- 12759330 TI - 2-[18F]F-A-85380: PET imaging of brain nicotinic acetylcholine receptors and whole body distribution in humans. AB - Noninvasive imaging of nicotinic acetylcholine receptors (nAChRs) in the human brain in vivo is critical for elucidating the role of these receptors in normal brain function and in the pathogenesis of brain disorders. Here we report the first in vivo visualization of human brain areas containing nAChRs by using PET and 2-[18F]fluoro-3-(2(S)azetidinylmethoxy)pyridine (2-[18F]FA). We acquired scans from six healthy non-smoking volunteers after i.v. bolus administration of 2-[18F]FA (1.6 MBq/kg or 0.043 +/- 0.002 mCi/kg). This dose was sufficient for visualizing nAChRs in the thalamus up to 5 h after injection. There were no adverse effects associated with administration of no-carrier-added 2-[18F]FA (1.3 10 pmol/kg). Consistent with the distribution of nAChRs in human brain, accumulated radioactivity was greatest in thalamus, intermediate in the midbrain, pons, cerebellum, and cortex; and least in white matter. As approximately 90% of the injected radioactivity was eliminated via the urine (biological half-life ca. 4 h), the urinary bladder wall received the highest radiation dose. The estimate of radiation dose equivalent to the urinary bladder wall (ca. 180 +/- 30 mSv/MBq or 0.7 rem/mCi with a 2.4 h void interval) suggests that multiple studies could be performed in a single subject. The results predict that quantitative PET imaging of nAChRs in human brain with 2-[18F]FA is feasible. PMID- 12759331 TI - How estrogen-specific proteins discriminate estrogens from androgens: a common steroid binding site architecture. AB - Steroid hormones play an essential role in a wide range of physiological and pathological processes, such as growth, metabolism, aging, and hormone-sensitive cancers. Estrogens are no exception and influence growth, differentiation, and functioning of many target tissues, such as the mammary gland, uterus, hypothalamus, pituitary, bone, and liver. Although very similar in structure, each steroid class (i.e., estrogens, androgens, progestins, mineral corticoids, or glucocorticoids) is responsible for distinct physiological processes. To permit specific biological responses for a given steroid class, specific proteins are responsible for steroid bioactivation, action, and inactivation, yet they have low or no affinity to other classes. Estrogens make no exception and possess their own set of related proteins. To understand the molecular basis underlying estrogen recognition from other steroids, structural features of estrogen specific proteins were analyzed along with their ability to discriminate between steroid hormones belonging to different classes. Hence, the study of all estrogen specific proteins for which an atomic structure has been determined demonstrated that a common steroid-binding pocket architecture is shared by these proteins. This architecture is composed of the following elements: i) a glutamate residue acting as a proton acceptor coupled with a proton donor that interact with the steroid O3; ii) a proton donor (His or Ser) that interacts with O17; iii) a highly conserved sandwich-like structure providing steric hindrance and preventing C19 steroid from binding; and iv) several amino acid residues interacting with the C18. As these different estrogen-specific proteins are not related in overall sequence, the inference is that the steroid binding site in these proteins has originated by convergent evolution. PMID- 12759332 TI - Nitric oxide induces polarization of actin in encephalitogenic T cells and inhibits their in vitro trans-endothelial migration in a p70S6 kinase-independent manner. AB - Nitric oxide (NO) inhibits both actively induced and transferred autoimmune encephalomyelitis. To explore potential mechanisms, we examined the ability of NO to inhibit migration of T lymphoblasts through both collagen matrices and monolayers of rat brain endothelial cells. The NO donor 1-hydroxy-2-oxo-3, 3-bis (2-aminoethyl)-1-triazene (HOBAT) inhibited migration in a concentration dependent manner. NO pretreatment of T cells inhibited migration through untreated endothelial cells, but NO pretreatment of endothelial cells had no inhibitory effect on untreated T cells. Therefore NO's migration inhibitory action was mediated through its effect on T cells and not endothelial cells. HOBAT did not inhibit migration by inducing T-cell death but rather by polarizing the T cells, resulting in a morphology suggestive of migrating cells. P70S6 kinase, shown to have a role in NO-induced migration inhibition in fibroblasts, had no role in the inhibitory effect of NO on T-cell migration. Thus, HOBAT did not alter p70S6K activity nor did rapamycin, a specific inhibitor of p70S6K, inhibit HOBAT-induced T-cell morphological changes or T-cell migration. We suggest that NO-induced morphological changes result in T cells with predefined migratory directionality, thus limiting the ability of these cells to respond to other migratory signals. PMID- 12759333 TI - HMGB1 inhibits cell death in yeast and mammalian cells and is abundantly expressed in human breast carcinoma. AB - Apoptosis is a fundamental biological process used to eliminate unwanted cells in a multicellular organism. An increasing number of regulatory proteins have been identified that either promote or inhibit apoptosis. For tumors to arise, apoptosis must be blocked in the transformed cells, for example by mutational overexpression of anti-apoptotic proteins, which represent attractive target proteins for molecular therapy strategies. In a functional yeast survival screen designed to select new anti-apoptotic mammalian genes, we have identified the chromosomal high-mobility group box-1 protein (HMGB1) as an inhibitor of yeast cell death induced by the pro-apoptotic Bcl-2 family member Bak. The C-terminal 33 amino acids of HMGB1 are dispensable for this inhibitory function. HMGB1 is also able to protect mammalian cells against different death stimuli including ultraviolet radiation, CD95-, TRAIL-, Casp-8-, and Bax-induced apoptosis. We found high HMGB1 protein levels in human primary breast carcinoma. Hmgb1 RNA levels are changing during different stages of mouse mammary gland development and are particularly low during lactation and involution. These data suggest that HMGB1 may participate in the regulation of mammary gland apoptosis and that its high expression level promotes tumor growth because of its anti-apoptotic properties. PMID- 12759334 TI - Tetracycline-dependent regulation of formamidopyrimidine DNA glycosylase in transgenic mice conditionally reduces oxidative DNA damage in vivo. AB - 8-Oxo-deoxyguanosine (8-oxo-dG) is a pervasive oxidative DNA lesion formed by endogenous oxidative stress and enhanced by drugs and environmental chemicals. This lesion results in transcriptional errors and mutations and is linked to neurodegeneration, teratogenesis, cancer, and other pathologies. We demonstrate that the neonatal central nervous system of transgenic mice carrying the tetracycline-regulable DNA repair gene formamidopyrimidine DNA glycosylase (fpg) has a 50% reduction in 8-oxo-dG levels. This enhanced DNA repair is suppressed by treatment with doxycycline. For the first time, this murine model permits the level of a specific DNA oxidation product to be regulated in a temporally and spatially specific manner, allowing its role as a primary or secondary factor in neurodegenerative disease to be determined in vivo. PMID- 12759335 TI - Identification of beta-carotene 15, 15'-monooxygenase as a peroxisome proliferator-activated receptor target gene. AB - Beta-carotene 15,15'-monooxygenase (BCM) catalyzes the first step of vitamin A biosynthesis from provitamin A carotenoids. We wished to determine the factors underlying the transcriptional regulation of this gene. After cloning of the 40 kilobase pair (kbp) mouse Bcm gene and determination of its genomic organization, analysis of the 2 kb 5'-flanking region showed several putative transcription factor binding sites including TATA box, a peroxisome proliferator response element (PPRE), AP2, and bHLH. The 2 kb fragment drove specific luciferase gene expression in vitro only in cell lines that express BCM (TC7, PF11, and monkey retinal pigment epithelium). Nucleotides -41 to +163, and -60 to +163 drove basal and specific Bcm transcriptional activity, respectively. Site-directed mutagenesis and gel shift experiments demonstrate that PPRE was essential for Bcm promoter specificity and that the peroxisome proliferator activated receptor (PPAR) gamma (PPARgamma) specifically binds to this element. Furthermore, cotransfection experiments and pharmacological treatments in vitro, using the specific PPARgamma agonists LY17883 and ciglitazone, demonstrate that the PPRE element confers peroxisome proliferator responsiveness via the PPARgamma and retinoid X receptor-alpha heterodimer. Treatment of mice with the PPARalpha/gamma agonist WY14643 increases BCM protein expression in liver. Thus PPAR is a key transcription factor for the transcriptional regulation of the Bcm gene, suggesting a broader function for PPARs in the regulation of carotenoid metabolism metabolism that is consistent with their established role in neutral lipid metabolism and transport. PMID- 12759336 TI - Novel stimulatory actions of the phytoestrogen genistein: effects on the gain of cardiac excitation-contraction coupling. AB - Genistein, a phytoestrogen found abundantly in soy products, is thought to be cardioprotective, partly through its ability to act as a natural Ca2+ channel antagonist. However, the precise nature and significance of such direct cardiac actions remain obscure. We investigated the hypothesis that genistein exerts important additional actions on cardiac excitation-contraction coupling (ECC). Genistein acutely increased cell shortening and the Ca2+ transient in field stimulated guinea-pig ventricular myocytes despite potently inhibiting the L-type Ca2+ current, I(Ca,L). The specific phosphotyrosine phosphatase inhibitor, bpV(phen), diminished the stimulatory effects of genistein on myocyte contractility, suggesting that the mechanism partly involved tyrosine kinase inhibition. Genistein increased sarcoplasmic reticulum (SR) Ca2+ load as measured with a caffeine pulse in Na+-free/ Ca2+-free solution. Furthermore, in the continued presence of caffeine, genistein increased the time constant of decline of the caffeine-induced Ca2+ transient, implying impaired sarcolemmal Na+/Ca2+ exchanger function. Tetanization studies in intact myocytes revealed that 43% of cells exhibited increased myofilament Ca2+ sensitivity in the presence of genistein. These findings demonstrate novel cardiac actions of genistein on the SR Ca2+ load, Na+/Ca2+ exchanger, and myofilament Ca2+ sensitivity, which result in an overall increase in myocyte contractility and consequently the gain of ECC. PMID- 12759337 TI - The terminal complement complex inhibits apoptosis in vascular smooth muscle cells by activating an autocrine IGF-1 loop. AB - Two counteracting processes determine accumulation of human vascular smooth muscle cells (SMCs) in atherosclerotic lesions: cell proliferation and apoptosis. SMCs synthesize insulin-like growth factor-1 (IGF-1), which potently inhibits apoptosis. The terminal complement complex C5b-9 interacts with SMCs in early human atherogenesis. In this study, we investigated whether C5b-9 may activate the IGF-1 system in SMCs, resulting in the inhibition of SMC apoptosis. C5b-9 generation on SMCs in vitro markedly reduced CD95-mediated apoptosis as assessed by flowcytometric analysis of annexin V binding and in caspase 3 assays. C5b-9 induced both significant IGF-1 release and up-regulation of IGF-1 binding sites in SMCs. Immunoneutralization of IGF-1 with a monoclonal IGF-1 antibody abolished the antiapoptotic effects of C5b-9. We conclude that C5b-9 inhibits apoptosis in SMCs by inducing an autocrine IGF-1 loop. This mechanism may contribute to the accumulation of SMCs in early human atherosclerotic lesions. PMID- 12759338 TI - Activation of c-Jun-N-terminal-kinase is crucial for the induction of a cell cycle arrest in human colon carcinoma cells caused by flurbiprofen enantiomers. AB - The unselective cyclooxygenase (COX) inhibitor S-flurbiprofen and its-in terms of COX-inhibition-"inactive" enantiomer R-flurbiprofen have been previously found to inhibit tumor development and growth in various animal models. The underlying mechanisms are unknown. Here, we show that both R- and S-flurbiprofen reduce survival of three colon cancer cell lines, which differ in the expression of COX 2 (HCT-15, no COX-2; Caco-2, inducible COX-2; and HT-29, constitutive COX-2). The IC50 for S- and R-flurbiprofen ranged from 250 to 450 microM. Both flurbiprofen enantiomers induced apoptosis in all three cell lines as indicated by DNA- and PARP-cleavage. In addition, R- and S-flurbiprofen caused a G1-cell cycle block. The latter was associated with an activation of c-Jun N-terminal kinase (JNK), an increase of the DNA binding activity of the transcription factor AP-1 and down regulation of cyclin D1 expression. Western blot analysis, as well as supershift experiments, revealed that the AP-1 activation was associated with a change of AP 1 composition toward an increase of JunB. The JNK inhibitor SP600125 antagonized R- and S-flurbiprofen-induced AP-1 DNA binding, suppression of cyclin D1 expression, and the G1-cell cycle block. However, JNK inhibition had no effect on flurbiprofen-induced apoptosis. Hence, the cell cycle arrest is obviously mediated, at least in part, through JNK-activation, whereas R- and S-flurbiprofen induced apoptosis is largely independent of JNK. Although in vitro effects of R- and S-flurbiprofen were indistinguishable, only R-flurbiprofen inhibited HCT-15 tumor growth in nude mice, suggesting the involvement of additional in vivo targets, which are differently affected by R- and S-flurbiprofen. PMID- 12759339 TI - Hoxb13 knockout adult skin exhibits high levels of hyaluronan and enhanced wound healing. AB - In contrast to adult cutaneous wound repair, early gestational fetal cutaneous wounds heal by a process of regeneration, resulting in little or no scarring. Previous studies indicate that down-regulation of HoxB13, a member of the highly conserved family of Hox transcription factors, occurs during fetal scarless wound healing. No down-regulation was noted in adult wounds. Here, we evaluate healing of adult cutaneous wounds in Hoxb13 knockout (KO) mice, hypothesizing that loss of Hoxb13 in adult skin should result in enhanced wound healing. Tensiometry was used to measure the tensile strength of incisional wounds over a 60-day time course; overall, Hoxb13 KO wounds are significantly stronger than wild-type (WT). Histological evaluation of incisional wounds shows that 7-day-old Hoxb13 KO wounds are significantly smaller and that 60-day-old Hoxb13 KO wounds exhibit a more normal collagen architecture compared with WT wounds. We also find that excisional wounds close at a faster rate in Hoxb13 KO mice. Biochemical and histochemical analyses show that Hoxb13 KO skin contains significantly elevated levels of hyaluronan. Because higher levels of hyaluronan and enhanced wound healing are characteristics of fetal skin, we conclude that loss of Hoxb13 produces a more "fetal-like" state in adult skin. PMID- 12759340 TI - Ischemic preconditioning preserves connexin 43 phosphorylation during sustained ischemia in pig hearts in vivo. AB - During myocardial ischemia, connexin 43 (Cx43) is dephosphorylated in vitro, and the subsequent opening of gap junctions formed by two opposing Cx43 hexamers was suggested to propagate ischemia/reperfusion injury. Reduction of infarct size (IS) by ischemic preconditioning (IP) involves activation of protein kinase C (PKC) and p38 mitogen activated protein kinase (MAPK), both of which can phosphorylate Cx43. We now studied in anesthetized pigs whether IP impacts on Cx43 phosphorylation by measuring the density of non-phosphorylated and total Cx43 (confocal laser) during normoperfusion and 90-min ischemia in non preconditioned and preconditioned hearts. Co-localization of PKCalpha, p38MAPKalpha, and p38MAPKbeta with Cx43 and the activity of p38MAPK were assessed. IP by 10 min ischemia and 15 min reperfusion reduced IS. Non phosphorylated Cx43 remained unchanged during ischemia in preconditioned hearts, while it increased from 35+/-3 to 75+/-8 AU (P<0.05) in non-preconditioned hearts. Co-localization of PKCalpha, p38MAPKalpha, and p38MAPKbeta with Cx43 during ischemia increased only in preconditioned hearts. While the ischemia induced increase in p38MAPKalpha activity was comparable in preconditioned and non-preconditioned hearts, p38MAPKbeta activity was increased only in preconditioned hearts. Blockade of p38MAPK by SB203580 attenuated the IS reduction and the increased p38MAPK-Cx43 co-localization by IP. We conclude that IP increases co-localization of protein kinases with Cx43 and preserves phosphorylation of Cx43 during ischemia. PMID- 12759341 TI - Soluble purine-converting enzymes circulate in human blood and regulate extracellular ATP level via counteracting pyrophosphatase and phosphotransfer reactions. AB - Extracellular ATP and other purines play a crucial role in the vasculature, and their turnover is selectively governed by a network of ectoenzymes expressed both on endothelial and hematopoietic cells. By studying the whole pattern of purine metabolism in human serum, we revealed the existence of soluble enzymes capable of both inactivating and transphosphorylating circulating purines. Evidence for this was obtained by using independent assays, including chromatographic analyses with 3H-labeled and unlabeled nucleotides and adenosine, direct transfer of gamma terminal phosphate from [gamma-32P]ATP to NDP/AMP, and bioluminescent measurement of ATP metabolism. Based on substrate-specificity and competitive studies, we identified three purine-inactivating enzymes in human serum, nucleotide pyrophosphatase (EC 3.6.1.9), 5'-nucleotidase (EC 3.1.3.5), and adenosine deaminase (EC 3.5.4.4), whereas an opposite ATP-generating pathway is represented by adenylate kinase (EC 2.7.4.3) and NDP kinase (EC 2.7.4.6). Comparative kinetic analysis revealed that the Vmax values for soluble nucleotide kinases significantly exceed those of counteracting nucleotidases, whereas the apparent Km values for serum enzymes were fairly comparable and varied within a range of 40-70 micro mol/l. Identification of soluble enzymes contributing, along with membrane-bound ectoenzymes, to the active cycling between circulating ATP and other purines provides a novel insight into the regulatory mechanisms of purine homeostasis in the blood. PMID- 12759342 TI - The neuropeptide galanin augments lobuloalveolar development. AB - Mammary lobuloalveolar development during pregnancy is controlled by ovarian sex steroids and pituitary prolactin release. In organ culture these hormones are incapable of reproducing the density and size of lobuloalveoli seen in mice, suggesting the existence of other undiscovered factors. We showed previously that galanin knockout mice fail to lactate sufficiently for pup survival following their first pregnancy. Here we demonstrate that prolactin treatment of galanin knockout mice allows pup survival but does not completely rescue lobuloalveolar development or reduced milk protein expression. When galanin was used in combination with prolactin in mammary organ culture, larger and more numerous lobules were produced than with prolactin alone. Galanin alone produced sustained activation of STAT5a and the induction of milk protein expression but did not induce lobulogenesis. Examination of the transcriptional interaction between galanin and prolactin using oligonucleotide microarrays demonstrated synergistic and antagonistic modes of interaction between these hormones. These data establish a new role for galanin as a hormone augmenting mammary development during pregnancy in concert with prolactin. PMID- 12759343 TI - Analysis of steady-state protein phosphorylation in mitochondria using a novel fluorescent phosphosensor dye. AB - The phosphorylation of mitochondrial proteins is pivotal to the regulation of respiratory activity in the cell and to signaling pathways leading to apoptosis, as well as for other vital mitochondrial processes. A number of protein kinases have been identified in mitochondria but the physiological substrates for many of these remain unknown or poorly understood. By necessity, most studies of mitochondrial phosphoproteins to date have been conducted using in vitro incorporation of 32P. However, proteins that are highly phosphorylated from in situ reactions are not necessarily detected by this approach. In this study, a new small molecule fluorophore has been employed to characterize steady-state levels of mitochondrial phosphoproteins. The dye is capable of sensitive detection of phosphorylated amino acid residues in proteins separated by gel electrophoresis. When the fluorescent dye is combined with a total protein stain in a sequential gel staining procedure, the phosphorylated proteins can be visualized in the same gel as the total proteins. To optimize resolution of the proteins in mitochondria, a previously described sucrose gradient fractionation method was employed prior to gel electrophoresis. Phosphorylated proteins, as defined by the fluorescence of the phosphosensor, were excised from the gels and identified by peptide mass fingerprinting. One novel and prominent phosphoprotein identified in this manner was determined to be the 42-kDa subunit of mitochondrial complex I. PMID- 12759344 TI - Physical and functional interactions between PML and MDM2. AB - The tumor suppressor protein PML and oncoprotein MDM2 have opposing effects on p53. PML stimulates p53 activity by recruiting it to nuclear foci termed PML nuclear bodies. In contrast, MDM2 inhibits p53 by promoting its degradation. To date, neither a physical nor functional relationship between PML and MDM2 has been described. In this study, we report an in vivo and in vitro interaction between PML and MDM2 which is independent of p53. Two separate regions of PML are recognized which can interact with MDM2. The C-terminal half of PML, encoded by residues 300-633, can interact with the central region of MDM2 which includes the MDM2 acidic domain. In addition, PML amino acids 1-200, which encode the RING finger and most of the B box zinc binding motifs, can interact with the C terminal, RING-finger containing region of MDM2. Interestingly, PML mutants in which sumoylation at lysine 160 was inhibited displayed an increased association with MDM2, suggesting that sumoylation at this site may be a determinant of PML MDM2 binding. Coexpression with MDM2 caused a redistribution of PML from the nucleus to the cytoplasm, and this required the PML N terminus and the MDM2 RING finger domain. These results suggest that interaction between the PML N terminus and MDM2 C terminus can promote PML nuclear exclusion. Wild-type MDM2 inhibited the ability of PML to stimulate the transcriptional activity of a GAL4-CBP fusion protein. This inhibition required the central, acidic region of MDM2, but did not require the MDM2 C terminus. Taken together, these studies demonstrate that MDM2 and PML can interact through at least two separate protein regions, and that these interactions can have specific effects on the activity and/or localization of PML. PMID- 12759345 TI - Isolation and characterization of a cone snail protease with homology to CRISP proteins of the pathogenesis-related protein superfamily. AB - The pathogenesis-related (PR) protein superfamily is widely distributed in the animal, plant, and fungal kingdoms and is implicated in human brain tumor growth and plant pathogenesis. The precise biological activity of PR proteins, however, has remained elusive. Here we report the characterization, cloning and structural homology modeling of Tex31 from the venom duct of Conus textile. Tex31 was isolated to >95% purity by activity-guided fractionation using a para nitroanilide substrate based on the putative cleavage site residues found in the propeptide precursor of conotoxin TxVIA. Tex31 requires four residues including a leucine N-terminal of the cleavage site for efficient substrate processing. The sequence of Tex31 was determined using two degenerate PCR primers designed from N terminal and tryptic digest Edman sequences. A BLAST search revealed that Tex31 was a member of the PR protein superfamily and most closely related to the CRISP family of mammalian proteins that have a cysteine-rich C-terminal tail. A homology model constructed from two PR proteins revealed that the likely catalytic residues in Tex31 fall within a structurally conserved domain found in PR proteins. Thus, it is possible that other PR proteins may also be substrate specific proteases. PMID- 12759346 TI - Hypochlorous acid generated by myeloperoxidase modifies adjacent tryptophan and glycine residues in the catalytic domain of matrix metalloproteinase-7 (matrilysin): an oxidative mechanism for restraining proteolytic activity during inflammation. AB - Dysregulation of matrix metalloproteinase (MMP) activity is implicated in tissue destruction under inflammatory conditions. An important mechanism controlling enzymatic activity might involve reactive oxygen species generated by phagocytes. Myeloperoxidase, a heme protein secreted by neutrophils, monocytes, and macrophages, uses hydrogen peroxide to generate hypochlorous acid (HOCl). We demonstrate that HOCl inhibits the activity of human matrilysin (MMP-7) in vitro, suggesting that it might limit proteolytic activity during inflammation. When MMP 7 was exposed to HOCl generated by myeloperoxidase, the proteinase lost activity. High performance liquid chromatographic analysis of the tryptic digest of the HOCl-treated proteinase demonstrated the absence of two peptides that were present in the untreated enzyme. Tandem mass spectrometric analysis revealed that both of the lost peptides contained methionine and tryptophan-glycine residues. The methionine residue of one of the peptides had been oxidized to methionine sulfoxide. In contrast, the major product from the other peptide was 4 atomic mass units smaller than its precursor (WG-4). This novel oxidation product was derived though modification of adjacent tryptophan and glycine residues in the catalytic domain of the enzyme. Loss of proteolytic activity was associated with conversion of the precursor peptide to WG-4 but not with methionine oxidation. In contrast, hydrogen peroxide failed to oxidize MMP-7 or to inactivate the enzyme. Thus, HOCl inactivates MMP-7, perhaps by site-specific conversion of tryptophan glycine to WG-4. This inactivation mechanism is distinct from the well studied mechanisms involving tissue inhibitors of metalloproteinases. Our findings suggest that local pericellular production of HOCl by phagocytes is a physiological mechanism for governing MMP activity during inflammation. PMID- 12759347 TI - Functional domains of chicken mitochondrial transcription factor A for the maintenance of mitochondrial DNA copy number in lymphoma cell line DT40. AB - Nuclear and mitochondrial (mt) forms of chicken mt transcription factor A (c TFAM) generated by alternative splicing of a gene (c-tfam) were cloned. c-tfam mapped at 6q1.1-q1.2 has similar exon/intron organization as mouse tfam except that the first exons encoding the nuclear and the mt form-specific sequences were positioned oppositely. When cDNA encoding the nuclear form was transiently expressed in chicken lymphoma DT40 cells after tagging at the C terminus with c Myc, the product was localized into nucleus, whereas the only endogenous mt form of DT40 cells was immunostained exclusively within mitochondria. c-TFAM is most similar to Xenopus (xl-) TFAM in having extended C-terminal regions in addition to two high mobility group (HMG) boxes, a linker region between them, and a C terminal tail, also found in human and mouse TFAM. Similarities between c- and xl TFAM are higher in linker and C-terminal regions than in HMG boxes. Disruption of both tfam alleles in DT40 cells prevented proliferation. The tfam+/tfam- cells showed a 50 and 40-60% reduction of mtDNA and its transcripts, respectively. Expression of exogenous wild type c-tfam cDNA in the tfam+/tfam- cells increased mtDNA up to 4-fold in a dose-dependent manner, whereas its transcripts increased only marginally. A deletion mutant lacking the first HMG box lost this activity, whereas only marginal reduction of the activity was observed in a deletion mutant at the second HMG box. Despite the essential role of the C-terminal tail in mtDNA transcription demonstrated in vitro, deletion of c-TFAM at this region reduced the activity of maintenance of the mtDNA level only by 50%. A series of deletion mutant at the tail region suggested stimulatory and suppressive sequences in this region for the maintenance of mtDNA level. PMID- 12759348 TI - Intracellular Na+ regulates dopamine and angiotensin II receptors availability at the plasma membrane and their cellular responses in renal epithelia. AB - The balance and cross-talk between natruretic and antinatruretic hormone receptors plays a critical role in the regulation of renal Na+ homeostasis, which is a major determinant of blood pressure. Dopamine and angiotensin II have antagonistic effects on renal Na+ and water excretion, which involves regulation of the Na+,K+-ATPase activity. Herein we demonstrate that angiotensin II (Ang II) stimulation of AT1 receptors in proximal tubule cells induces the recruitment of Na+,K+-ATPase molecules to the plasmalemma, in a process mediated by protein kinase Cbeta and interaction of the Na+,K+-ATPase with adaptor protein 1. Ang II stimulation led to phosphorylation of the alpha subunit Ser-11 and Ser-18 residues, and substitution of these amino acids with alanine residues completely abolished the Ang II-induced stimulation of Na+,K+-ATPase-mediated Rb+ transport. Thus, for Ang II-dependent stimulation of Na+,K+-ATPase activity, phosphorylation of these serine residues is essential and may constitute a triggering signal for recruitment of Na+,K+-ATPase molecules to the plasma membrane. When cells were treated simultaneously with saturating concentrations of dopamine and Ang II, either activation or inhibition of the Na+,K+-ATPase activity was produced dependent on the intracellular Na+ concentration, which was varied in a very narrow physiological range (9-19 mm). A small increase in intracellular Na+ concentrations induces the recruitment of D1 receptors to the plasma membrane and a reduction in plasma membrane AT1 receptors. Thus, one or more proteins may act as an intracellular Na+ concentration sensor and play a major regulatory role on the effect of hormones that regulate proximal tubule Na+ reabsorption. PMID- 12759349 TI - A flux model of glycolysis and the oxidative pentosephosphate pathway in developing Brassica napus embryos. AB - Developing oilseeds synthesize large quantities of triacylglycerol from sucrose and hexose. To understand the fluxes involved in this conversion, a quantitative metabolic flux model was developed and tested for the reaction network of glycolysis and the oxidative pentose phosphate pathway (OPPP). Developing Brassica napus embryos were cultured with [U-13C6]glucose, [1-13C]glucose, [6 13C]glucose, [U-13C12]sucrose, and/or [1,2-13C2]glucose and the labeling patterns in amino acids, lipids, sucrose, and starch were measured by gas chromatography/mass spectrometry and NMR. Data were used to verify a reaction network of central carbon metabolism distributed between the cytosol and plastid. Computer simulation of the steady state distribution of isotopomers in intermediates of the glycolysis/OPPP network was used to fit metabolic flux parameters to the experimental data. The observed distribution of label in cytosolic and plastidic metabolites indicated that key intermediates of glycolysis and OPPP have similar labeling in these two compartments, suggesting rapid exchange of metabolites between these compartments compared with net fluxes into end products. Cycling between hexose phosphate and triose phosphate and reversible transketolase velocity were similar to net glycolytic flux, whereas reversible transaldolase velocity was minimal. Flux parameters were overdetermined by analyzing labeling in different metabolites and by using data from different labeling experiments, which increased the reliability of the findings. Net flux of glucose through the OPPP accounts for close to 10% of the total hexose influx into the embryo. Therefore, the reductant produced by the OPPP accounts for at most 44% of the NADPH and 22% of total reductant needed for fatty acid synthesis. PMID- 12759350 TI - Role of hexosamine biosynthesis in glucose-mediated up-regulation of lipogenic enzyme mRNA levels: effects of glucose, glutamine, and glucosamine on glycerophosphate dehydrogenase, fatty acid synthase, and acetyl-CoA carboxylase mRNA levels. AB - Glucose uptake into adipose and liver cells is known to up-regulate mRNA levels for various lipogenic enzymes such as fatty acid synthase (FAS) and acetyl-CoA carboxylase (ACC). To determine whether the hexosamine biosynthesis pathway (HBP) mediates glucose regulation of mRNA expression, we treated primary cultured adipocytes for 18 h with insulin (25 ng/ml) and either glucose (20 mm) or glucosamine (2 mm). A ribonuclease protection assay was used to quantitate mRNA levels for FAS, ACC, and glycerol-3-P dehydrogenase (GPDH). Treatment with insulin and various concentrations of d-glucose increased mRNA levels for FAS (280%), ACC (93%), and GPDH (633%) in a dose-dependent manner (ED50 8-16 mm). Mannose similarly elevated mRNA levels, but galactose and fructose were only partially effective. l-glucose had no effect. Omission of glutamine from the culture medium markedly diminished the stimulatory effect of glucose on mRNA expression. Since glutamine is a crucial amide donor in hexosamine biosynthesis, we interpret these data to mean that glucose flux through the HBP is linked to regulation of lipogenesis through control of gene expression. Further evidence for hexosamine regulation was obtained using glucosamine, which is readily transported into adipocytes where it directly enters the HBP. Glucosamine was 15 30 times more potent than glucose in elevating FAS, ACC, and GPDH mRNA levels (ED50 approximately 0.5 mm). In summary: 1) GPDH, FAS, and ACC mRNA levels are upregulated by glucose; 2) glucose-induced up-regulation requires glutamine; and 3) mRNA levels for lipogenic enzymes are up-regulated by glucosamine. Hyperglycemia is the hallmark of diabetes mellitus and leads to insulin resistance, impaired glucose metabolism, and dyslipidemia. We postulate that disease pathophysiology may have a common underlying factor, excessive glucose flux through the HBP. PMID- 12759351 TI - Phosphorylation at serine 75 is required for UV-mediated degradation of human Cdc25A phosphatase at the S-phase checkpoint. AB - The human Cdc25A phosphatase plays a pivotal role at the G1/S transition by activating cyclin E and A/Cdk2 complexes through dephosphorylation. In response to ionizing radiation, Cdc25A is phosphorylated by both Chk1 and Chk2 on Ser-123. This in turn leads to ubiquitylation and rapid degradation of Cdc25A by the proteasome resulting in cell cycle arrest. We found that in response to UV irradiation, Cdc25A is phosphorylated at a different serine residue, Ser-75. Significantly, Cdc25A mutants carrying alanine instead of either Ser-75 or Ser 123 demonstrate that only Ser-75 mediates protein stabilization in response to UV induced DNA damage. As a consequence, cyclin E/Cdk2 kinase activity was high. Furthermore, we find that Cdc25A was phosphorylated by Chk1 on Ser-75 in vitro and that the same site was also phosphorylated in vivo. Taken together, these data strongly suggest that phosphorylation of Cdc25A on Ser-75 by Chk1 and its subsequent degradation is required to delay cell cycle progression in response to UV-induced DNA lesions. PMID- 12759352 TI - The amino-terminal GAF domain of Azotobacter vinelandii NifA binds 2-oxoglutarate to resist inhibition by NifL under nitrogen-limiting conditions. AB - The expression of genes required for the synthesis of molybdenum nitrogenase in Azotobacter vinelandii is controlled by the NifL-NifA transcriptional regulatory complex in response to nitrogen, carbon, and redox status. Activation of nif gene expression by the transcriptional activator NifA is inhibited by direct protein protein interaction with NifL under conditions unfavorable for nitrogen fixation. We have recently shown that the NifL-NifA system responds directly to physiological concentrations of 2-oxoglutarate, resulting in relief of NifA activity from inhibition by NifL under conditions when fixed nitrogen is limiting. The inhibitory activity of NifL is restored under conditions of excess combined nitrogen through the binding of the signal transduction protein Av GlnK to the carboxyl-terminal domain of NifL. The amino-terminal domain of NifA comprises a GAF domain implicated in the regulatory response to NifL. A truncated form of NifA lacking this domain is not responsive to 2-oxoglutarate in the presence of NifL, suggesting that the GAF domain is required for the response to this ligand. Using isothermal titration calorimetry, we demonstrate stoichiometric binding of 2-oxoglutarate to both the isolated GAF domain and the full-length A. vinelandii NifA protein with a dissociation constant of approximately 60 microm. Limited proteolysis experiments indicate that the binding of 2-oxoglutarate increases the susceptibility of the GAF domain to trypsin digestion and also prevents NifL from protecting these cleavage sites. However, protection by NifL is restored when the non-modified (non-uridylylated) form of Av GlnK is also present. Our results suggest that the binding of 2 oxoglutarate to the GAF domain of NifA may induce a conformational change that prevents inhibition by NifL under conditions when fixed nitrogen is limiting. PMID- 12759353 TI - Processing of seminal plasma hCAP-18 to ALL-38 by gastricsin: a novel mechanism of generating antimicrobial peptides in vagina. AB - The human cathelicidin, hCAP-18, is expressed both in neutrophils and in epithelial cells. hCAP-18 is processed to the antimicrobial peptide LL-37 by proteinase 3 in neutrophils. hCAP-18 is highly expressed in the epididymis with a subsequent high concentration in seminal plasma where the protein is present in its unprocessed and antimicrobially inactive form. We report here that hCAP-18 in seminal plasma is processed to generate a 38-amino acid antimicrobial peptide ALL 38 by the prostate-derived protease gastricsin when incubated at a pH corresponding to the vaginal pH. In accordance with this, seminal plasma derived hCAP-18 was found in its processed form in the vagina following sexual intercourse. The antimicrobial activity of ALL-38 against a variety of microorganisms tested is equal to that of LL-37. This enzymatic activation of a proantimicrobial substance in seminal plasma following exposure to the vaginal milieu represents a novel mechanism to prevent infection following sexual intercourse. PMID- 12759354 TI - Activation of the p70 S6 kinase and phosphorylation of the 4E-BP1 repressor of mRNA translation by type I interferons. AB - The Type I IFN receptor-generated signals required for initiation of mRNA translation and, ultimately, induction of protein products that mediate IFN responses, remain unknown. We have previously shown that IFNalpha and IFNbeta induce phosphorylation of insulin receptor substrate proteins and downstream engagement of the phosphatidylinositol (PI) 3'-kinase pathway. In the present study we provide evidence for the existence of a Type I IFN-dependent signaling cascade activated downstream of PI 3'-kinase, involving p70 S6 kinase. Our data demonstrate that p70 S6K is rapidly phosphorylated on threonine 421 and serine 424 and is activated during treatment of cells with IFNalpha or IFNbeta. Such activation of p70 S6K is blocked by pharmacological inhibitors of the PI 3' kinase or the FKBP 12-rapamycin-associated protein/mammalian target of rapamycin (FRAP/mTOR). Consistent with this, the Type I IFN-dependent phosphorylation/activation of p70 S6K is defective in embryonic fibroblasts from mice with targeted disruption of the p85alpha and p85beta subunits of the PI 3' kinase (p85alpha-/-beta-/-). Treatment of sensitive cell lines with IFNalpha or IFNbeta also results in phosphorylation/inactivation of the 4E-BP-1 repressor of mRNA translation. Such 4E-BP1 phosphorylation is also PI3'-kinase-dependent and rapamycin-sensitive, indicating that the Type I IFN-inducible activation of PI3' kinase and FRAP/mTOR results in dissociation of 4E-BP1 from the eukaryotic initiation factor-4E (eIF4E) complex. Altogether, our data establish that the Type I IFN receptor-activated PI 3'-kinase pathway mediates activation of the p70 S6 kinase and inactivation of 4E-BP1, to regulate mRNA translation and induction of Type I IFN responses. PMID- 12759355 TI - The Cdk inhibitor p21 is required for necrosis, but it inhibits apoptosis following toxin-induced liver injury. AB - Liver injury and repair were examined in wild type, p21Waf1/Cip1, and p27Kip1 deficient mice following carbon tetrachloride (CCl4) administration. In wild type liver, p21 expression is induced in a biphasic manner following injection of CCl4, with an early peak of p21 expression occurring in pericentral hepatocytes at 6 h, prior to evidence of injury, and a second peak succeeding regenerative proliferation. In contrast, p27 is present throughout the quiescent liver, but its expression decreases following CCl4 injection. Surprisingly, p21-deficient animals were resistant to CCl4-induced necrotic injury, indicating that rapid induction of p21 in pericentral hepatocytes following CCl4 injection contributes to subsequent necrosis. Expression of cytochrome P450 2E1, which plays an essential role in CCl4-induced necrotic injury, was not affected in p21-deficient mice. Although they had the least injury, p21-deficient mice had the highest levels of hepatic proliferation that correlated with increases in hyperphosphorylated retinoblastoma protein and Cyclin A gene expression. Increased replication in p21-deficient livers was counteracted by an increase in hepatocyte apoptosis as detected by caspase-3 activation. p21 plays distinct and opposing roles regulating hepatocyte survival during injury and subsequent repair, with early induction of p21 contributing to necrotic injury and later expression to cessation of proliferation and hepatocyte survival. PMID- 12759357 TI - Two functional heads are required for full activation of smooth muscle myosin. AB - The motor activity of smooth muscle myosin II is regulated by the regulatory light chain phosphorylation, but it is not understood how phosphorylation activates motor activity. To address this question, we produced asymmetric heavy meromyosin (HMM), which is composed of a wild-type (WT) heavy chain and a mutant heavy chain having no motor activity (i.e. S236T or G457A). The actin-activated ATPase activities (Vmax) of asymmetric HMMs were only 21.8 and 8.4% of the wild type HMM for S236A/WT HMM and G456A/WT HMM, respectively. If the two heads of HMM are independent for their ATPase activities, asymmetric HMM should show 50% of the activity of wild-type HMM; however, the activity of asymmetric HMM was much lower than the expected value. The results suggest that the activity of the wild type head is attenuated by the presence of inactive head. Consistently, the actin gliding velocity of the asymmetric HMM (i.e. S236T/WT or G457A/WT) was less than one-fifth of the wild-type HMM. The present study supports an idea that the two heads of smooth muscle myosin II interact with each other and the presence of two active heads is required for full activation. PMID- 12759356 TI - NF-kappaB specifically activates BMP-2 gene expression in growth plate chondrocytes in vivo and in a chondrocyte cell line in vitro. AB - Bone morphogenetic protein-2 (BMP-2) regulates growth plate chondrogenesis during development and postnatal bone growth, but the control mechanisms of BMP-2 expression in growth plate chondrocytes are unknown. Here we have used both in vitro and in vivo approaches to demonstrate that transcription factor, NF-kappaB, regulates BMP-2 gene expression in chondrocytes. Two putative NF-kappaB response elements were found in the -2712/+165 region of the BMP-2 gene. Cotransfection of mutant I-kappaBalpha expression plasmids with BMP-2 promoter-luciferase reporters into TMC-23 chondrocyte cell line suppressed BMP-2 transcription. Mutations in NF kappaB response elements in the BMP-2 gene lead to decreases in BMP-2 promoter activity. Electrophoretic mobility shift assay using nuclear extracts from TMC-23 chondrocytic cells revealed that the NF-kappaB subunits p50 and p65 bound to the NF-kappaB response elements of the BMP-2 gene. Thus, NF-kappaB may positively regulate BMP-2 gene transcription. Consistent with these findings, expression of BMP-2 mRNA was significantly reduced in growth plate chondrocytes in NF-kappaB p50/p52 dKO mice, which associated with decreased numbers of 5-bromo-2' deoxyuridine (BrdUrd)-positive cells in the proliferating zone of growth plate in these mice. Therefore, in postnatal growth plate chondrocytes, expression of BMP 2 is regulated by NF-kappaB, which may play an important role in chondrogenesis. PMID- 12759358 TI - Bacterial injection machines. PMID- 12759359 TI - Recruitment of nectin-3 to cell-cell junctions through trans-heterophilic interaction with CD155, a vitronectin and poliovirus receptor that localizes to alpha(v)beta3 integrin-containing membrane microdomains. AB - Nectins present a novel class of Ig superfamily adhesion molecules that, cooperatively with cadherins, establish and maintain cell-cell adherens junctions. CD155, the cognate receptor for poliovirus, undergoes cell-matrix contacts by binding to the extracellular matrix protein vitronectin. The significant homology of nectins with CD155 prompted us to investigate the possibility of their interaction. We determined that nectin-3 binds CD155 and its putative mouse homologue Tage4 in cell-based ligand binding assays. Coculture of nectin-3- and CD155-expressing HeLa cells led to CD155-dependent recruitment of nectin-3 to cell-cell contacts. In a heterologous coculture system with CD155 expressing mouse neuroblastoma cells, HeLa cell-expressed nectin-3 was recruited to contact sites with CD155 bearing neurites. CD155 and nectin-3 colocalized to epithelial cell-cell junctions in renal proximal tubules and in the amniotic membrane. Efficient interaction depended on CD155 dimerization, which appears to be aided by cell type-specific cofactors. We furthermore found CD155 to codistribute with alpha(v) integrin microdomains on the surface of transfected mouse fibroblasts and at amniotic epithelial cell junctions. Our findings demonstrate the possible trans-interaction between the bona fide cell-cell adherens type adhesion system (cadherin/nectin) and the cell-matrix adhesion system (integrin/CD155) by virtue of their nectin-3 and CD155 components, respectively. PMID- 12759360 TI - Nanosecond dynamics of the mouse acetylcholinesterase cys69-cys96 omega loop. AB - The paradox of high substrate turnover occurring within the confines of a deep, narrow gorge through which acetylcholine must traverse to reach the catalytic site of acetylcholinesterase has suggested the existence of transient gorge enlargements that would enhance substrate accessibility. To establish a foundation for the experimental study of transient fluctuations in structure, site-directed labeling in conjunction with time-resolved fluorescence anisotropy were utilized to assess the possible involvement of the omega loop (Omega loop), a segment that forms the outer wall of the gorge. Specifically, the flexibility of three residues (L76C, E81C, and E84C) in the Cys69-Cys96 Omega loop and one residue (Y124C) across the gorge from the Omega loop were studied in the absence and presence of two inhibitors of different size, fasciculin and huperzine. Additionally, to validate the approach molecular dynamics was employed to simulate anisotropy decay of the side chains. The results show that the Omega loop residues are significantly more mobile than the non-loop residue facing the interior of the gorge. Moreover, fasciculin, which binds at the mouth of the gorge, well removed from the active site, decreases the mobility of 5-((((2 acetyl)amino)ethyl)amino)naphthalene-1-sulfonic acid reporter groups attached to L76C and Y124C but increases the mobility of the reporter groups attached to E81C and E84C. Huperzine, which binds at the base of active-site gorge, has no effect on the mobility of reporter groups attached to L76C and Y124C but increases the mobility of the reporter groups attached to E81C and E84C. Besides showing that fluctuations of the Omega loop residues are not tightly coupled, the results indicate that residues in the Omega loop exhibit distinctive conformational fluctuations and therefore are likely to contribute to transient gorge enlargements in the non-liganded enzyme. PMID- 12759361 TI - Deletion of N-terminal residues 23-88 from prion protein (PrP) abrogates the potential to rescue PrP-deficient mice from PrP-like protein/doppel-induced Neurodegeneration. AB - Accumulating evidence has suggested that prion protein (PrP) is neuroprotective and that a PrP-like protein/Doppel (PrPLP/Dpl) is neurotoxic. A line of PrP deficient mice, Ngsk Prnp0/0, ectopically expressing PrPLP/Dpl in neurons, exhibits late-onset ataxia because of Purkinje cell death that is prevented by a transgene encoding wild-type mouse PrP. To elucidate the mechanisms of neurodegeneration in these mice, we introduced five types of PrP transgene, namely one heterologous hamster, two mouse/hamster chimeric genes, and two mutants, each of which encoded PrP lacking residues 23-88 (MHM2.del23-88) or with E199K substitution (Mo.E199K), into Ngsk Prnp0/0 mice. Only MHM2.del23-88 failed to rescue the mice from the Purkinje cell death. The transgenic mice, MHM2.del23 88/Ngsk Prnp0/0, expressed several times more PrP than did wild-type (Prnp+/+) mice and PrPLP/Dpl at an equivalent level to Ngsk Prnp0/0 mice. Little difference was observed in the pathology and onset of ataxia between Ngsk Prnp0/0 and MHM2.del23-88/Ngsk Prnp0/0. No detergent-insoluble PrPLP/Dpl was detectable in the central nervous system of Ngsk Prnp0/0 mice even after the onset of ataxia. Our findings provide evidence that the N-terminal residues 23-88 of PrP containing the unique octapeptide-repeat region is crucial for preventing Purkinje cell death in Prnp0/0 mice expressing PrPLP/Dpl in the neuron. PMID- 12759362 TI - Identification and characterization of DEN1, a deneddylase of the ULP family. AB - To identify deneddylases, proteases with specificity for hydrolysis of Nedd8 derivatives, a facile method was developed for the synthesis of Nedd8 amidomethylcoumarin (a substrate) and Nedd8 vinyl sulfone (an inhibitor). Deneddylase activity is necessary to reverse the conjugation of Nedd8 to cullin, a modification that regulates at least some ubiquitin ligases. The reaction of Nedd8 vinyl sulfone with L-M(TK-) mouse fibroblast lysates identified two deneddylases. The deubiquitinating enzyme UCH-L3 is labeled by both ubiquitin vinyl sulfone and Nedd8 vinyl sulfone. In contrast, a second and more selective enzyme is labeled only by Nedd8 vinyl sulfone. This protein, DEN1, is a 221-amino acid thiol protease that is encoded by an open reading frame previously annotated as SENP8. Recombinant human DEN1 shows significant specificity for Nedd8 and catalyzes the hydrolysis of Nedd8 amidomethylcoumarin with a Km of 51 nm and a kcat of7s-1. The catalytic efficiency of DEN1 acting upon ubiquitin amidomethylcoumarin is 6 x 10-4 that of Nedd8 amidomethylcoumarin and its activity on SUMO-1 amidomethylcoumarin is undetectable. This selectivity was unexpected as DEN1 is most closely related to enzymes that catalyze desumoylation. This observation expands to four the number of DUB families with members that can process the C terminus of Nedd8. PMID- 12759363 TI - DEN1 is a dual function protease capable of processing the C terminus of Nedd8 and deconjugating hyper-neddylated CUL1. AB - Nedd8 activates ubiquitination by increasing the efficiency of polyubiquitin chain assembly through its covalent conjugation to cullin molecules. Here we report the isolation, cloning, and characterization of a novel human Nedd8 specific protease called DEN1. Human DEN1 is encoded by AAH31411.1, a previously uncharacterized protein of 212 amino acids that shares homology with the Ulp1 cysteinyl SUMO deconjugating enzyme family. Recombinant human DEN1, purified from bacteria, selectively binds to Nedd8 and hydrolyzes C-terminal derivatives of Nedd8. Interestingly, DEN1 deconjugates cullin 1 (CUL1)-Nedd8 in a concentration dependent manner. At a low concentration, DEN1 processes hyper-neddylated CUL1 to yield a mononeddylated form, which presumably contains the Lys-720CUL1-Nedd8 linkage. At elevated concentrations, DEN1 is able to complete the removal of Nedd8 from CUL1. These activities distinguish DEN1 from the COP9 signalosome, which is capable of efficiently cleaving the Lys-720CUL1-Nedd8 conjugate, but lacks Nedd8 C-terminal hydrolytic activity and poorly processes hyperneddylated CUL1. These results suggest a unique role for DEN1 in regulating the modification of cullins by Nedd8. PMID- 12759365 TI - The beta-subunit of the signal recognition particle receptor is a novel GTP binding protein without intrinsic GTPase activity. AB - The beta-subunit of the signal recognition particle receptor (SRbeta), a member of the Ras family of small molecular weight GTPases, is involved in the targeting of nascent polypeptide chains to the protein translocation machinery in the endoplasmic reticulum membrane. We purified SRbeta from an expressing strain of Escherichia coli and investigated the properties of the isolated GTPase. We find that, unlike other Ras family GTPases, most SRbeta purifies bound to GTP, and SRbeta-bound GTP is not easily exchanged with solution GTP. SRbeta possesses no detectable GTPase activity. Although a stable interaction between SRbeta and ribosomes is observed, SRbeta is not stimulated to hydrolyze GTP when incubated with ribosomes or ribosome-nascent chains. A GTPase mutant harboring a mutation in a region predicted to be functionally important, based on observations made in related GTPases, binds GTP with faster kinetics and appears to be a less stable protein but otherwise displays similar properties to the wild-type SRbeta GTPase. Our results demonstrate that as an isolated GTPase, SRbeta functions differently from the Arf- and Ras-type GTPases that it is most closely related to by sequence. PMID- 12759364 TI - Functional interaction of the DNA-binding transcription factor Sp1 through its DNA-binding domain with the histone chaperone TAF-I. AB - Transcription involves molecular interactions between general and regulatory transcription factors with further regulation by protein-protein interactions (e.g. transcriptional cofactors). Here we describe functional interaction between DNA-binding transcription factor and histone chaperone. Affinity purification of factors interacting with the DNA-binding domain of the transcription factor Sp1 showed Sp1 to interact with the histone chaperone TAF-I, both alpha and beta isoforms. This interaction was specific as Sp1 did not interact with another histone chaperone CIA nor did other tested DNA-binding regulatory factors (MyoD, NFkappaB, p53) interact with TAF-I. Interaction of Sp1 and TAF-I occurs both in vitro and in vivo. Interaction with TAF-I results in inhibition of DNA-binding, and also likely as a result of such, inhibition of promoter activation by Sp1. Collectively, we describe interaction between DNA-binding transcription factor and histone chaperone which results in negative regulation of the former. This novel regulatory interaction advances our understanding of the mechanisms of eukaryotic transcription through DNA-binding regulatory transcription factors by protein-protein interactions, and also shows the DNA-binding domain to mediate important regulatory interactions. PMID- 12759366 TI - Adjacent sequence controls the response polarity of nitric oxide-sensitive Sp factor binding sites. AB - Nitric oxide (NO*) and cAMP-dependent protein kinase (PKA) inhibitors up-regulate tumor necrosis factor alpha (TNFalpha) by decreasing Sp1 binding to a proximal GC box element. Here, elements flanking GC boxes were tested for their role in determining whether Sp sites act as activators or repressors. Promoter studies in receptive human cell lines demonstrated that NO* down-regulated endothelial NO* synthase (eNOS) but up-regulated TNFalpha. Like TNFalpha, Sp1 binding to the eNOS promoter was decreased by NO* and a PKA inhibitor, H89, and increased by a PKA activator, dibutyryl cAMP (Bt2cAMP). For either promoter, mutation of Sp sites abolished NO* responses. In contrast, mutation of an upstream AP1 site in the TNFalpha promoter (not present in eNOS) maintained NO* responsiveness, but reversed the direction of NO* and cAMP effects. Using artificial constructs, NO* increased transcription when Sp and AP1 sites were both present (TNFalpha-like response), but decreased it when the adjacent AP1 site was disrupted (eNOS-like response). NO*, H89, and Bt2cAMP were found to produce reciprocal protein binding changes at contiguous AP1 and Sp sites (p < 0.0001 for an interaction). Chromatin immunoprecipitation assays demonstrated that Sp1 and to a lesser extent Sp3 bound to the GC box regions of eNOS and TNFalpha in intact cells. Thus, this NO*- and cAMP-responsive regulatory module has a Sp site sensor variably coupled to an adjacent element that determines response polarity. These results define a composite element that can utilize secondary inputs to convert off signals to on, thereby conferring complex functionalities to the same DNA binding motif. PMID- 12759368 TI - Measuring antioxidants in tree species in the natural environment: from sampling to data evaluation. AB - Biochemical measurements of antioxidants and protective pigments have been successfully introduced as markers of environmental stress in field studies (mainly forest studies). A guideline for field sampling and analysis methods is required to allow better comparison of data from different studies. The present review paper recommends HPLC methods for the analysis of ascorbate and glutathione (in oxidized and reduced form), tocopherols, and chloroplast pigments. Methodological variations are substantially lower (coefficients of variance of repeated extractions typically 4-9%) than biological variations of field samples (typical variation coefficients 8-36%), hence special emphasis is put on considerations of sampling standardization in the field with respect to sample time (seasonal and diurnal) and representative sampling of individuals and tissues. Following the suggestions in this paper would enable researchers to produce results that could be compared with those of several forest studies on conifers published in recent years. A larger data-set available for multivariate statistical evaluations (e.g. principal component analysis and cluster analysis) will enhance the diagnostic value of such investigations. PMID- 12759369 TI - Ectopic expression of syntaxin 1 in the ER redirects TI-VAMP- and cellubrevin containing vesicles. AB - SNARE proteins are key mediators of membrane fusion. Their function in ensuring compartmental specificity of membrane fusion has been suggested by in vitro studies but not demonstrated in vivo. We show here that ectopic expression of the plasma membrane t-SNARE heavy chain syntaxin 1 in the endoplasmic reticulum induces the redistribution of its cognate vesicular SNAREs, TI-VAMP and cellubrevin, and its light chain t-SNARE SNAP-23. These effects were prevented by co-expressing nSec1. Expression of syntaxin 1 alone impaired the cell surface expression of TI-VAMP and cellubrevin but not the recycling of transferrin receptor. TI-VAMP, cellubrevin and SNAP-23 associated in vivo with exogenous syntaxin 1. Redistribution of TI-VAMP in the ER of syntaxin-1-expressing cells was microtubule dependent and impaired the trafficking of CD63, a cargo of TI VAMP-containing vesicles. We conclude that the destination of v-SNAREs is driven by their specific interaction with cognate t-SNAREs. Our in vivo data provide strong support for the theory that highly specific v-SNARE-t-SNARE interactions control compartmental specificity of membrane fusion. PMID- 12759370 TI - alpha3beta1-integrin regulates hair follicle but not interfollicular morphogenesis in adult epidermis. AB - alpha3beta1-integrin is abundantly expressed in the epidermis, and in mice, ablation of the alpha3 gene results in embryonic defects and perinatal lethality. To determine the role of alpha3-integrin in adult skin development, we grafted skin from newborn alpha3-integrin-deficient mice on to ICRF nu/nu recipients. We report that adult alpha3-integrin-deficient skin has severe abnormalities restricted to hair follicle morphology, which include stunted hair follicle growth, increased hair follicle fragility, aberrant pigment accumulation and formation of hair follicle clusters. These abnormalities are caused by a combination of defects in: (1) keratinocyte cytoskeletal organisation, (2) outer root sheath architecture and (3) integrity of the lamina densa. Our results indicate that alpha3beta1 is not essential for adult interfollicular epidermal differentiation, but it is required to direct several processes important in hair follicle maintenance and morphogenesis. PMID- 12759371 TI - Regulation of early endocytic vesicle motility and fission in a reconstituted system. AB - We previously established conditions to reconstitute kinesin-dependent early endocytic vesicle motility and fission on microtubules in vitro. The present study examined the question whether motility and fission are regulated in this system. Screening for proteins by immunofluorescence microscopy revealed that the small G protein, Rab4, was associated with 80% of hepatocyte-derived early endocytic vesicles that contain the ligand asialoorosomucoid (ASOR). By contrast, other markers for early endocytic vesicles including clathrin, Rab5 and EEA1 were present in the preparation but did not colocalize with the ASOR vesicles. Guanine nucleotides exchanged into the Rab4 present on the vesicles as shown by solubilization of Rab4 by Rab-GDI; solubilization was inhibited by incubation with GTP-gamma-S and promoted by GDP. Pre-incubation of vesicles with GDP increased the number of vesicles moving on microtubules and markedly increased vesicle fission. This increase in motility from GDP was shown to be towards the minus end of microtubules, possibly through activation of the minus-end-directed kinesin, KIFC2. Pre-incubation of vesicles with GTP-gamma-S, by contrast, repressed motility. Addition of exogenous GST-Rab4- GTP-gamma-S led to a further repression of motility and fission. Repression was not seen with addition of GST Rab4-GDP. Treatment of vesicles with Rab4 antibody also repressed motility, and repression was not seen when vesicles were pre-incubated with GDP. Based on these results we hypothesize that endogenous Rab4-GTP suppresses motility of ASOR containing vesicles in hepatocytes and that conversion of Rab4-GTP to Rab4-GDP serves as a molecular switch that activates minus-end kinesin-based motility, facilitating early endosome fission and consequent receptor-ligand segregation. PMID- 12759372 TI - The type 3 inositol 1,4,5-trisphosphate receptor is concentrated at the tight junction level in polarized MDCK cells. AB - The subcellular localization of inositol 1,4,5-trisphosphate (InsP3)-induced Ca2+ signals is important for the activation of many physiological functions. In epithelial cells the spatial distribution of InsP3 receptor is restricted to specific areas, but little is known about the relationship between the receptor's distribution and cell polarity. To investigate this relationship, the best known polarized cell model, MDCK, was examined. This cell line is characterized by a strong expression of the type 3 InsP3 receptor and the subcellular localization of this receptor was followed during cell polarization using immunofluorescence and confocal analysis. In non-polarized cells, including ras transformed f3 MDCK cells, the type 3 InsP3 receptor was found to co-localize with markers of the endoplasmic reticulum in the cytoplasm. In contrast, in polarized cells, this receptor was mostly distributed at the apex of the lateral plasma membrane with the markers of tight junctions, ZO-1 and occludin. The localization of the type 3 InsP3 receptor in the vicinity of tight junctions was confirmed by immunogold electron microscopy. The culture of MDCK cells in calcium-deprived medium, led to disruption of cell polarity and receptor redistribution in the cytoplasm. Addition of calcium to these deprived cells induced the restoration of polarity and the relocalization of the receptor to the plasma membrane. MDCK cells were stably transfected with a plasmid coding the full-length mouse type 1 InsP3 receptor tagged with EGFP at the C-terminus. The EGFP-tagged type 1 receptor and the endogenous type 3 co-localized in the cytoplasm of non-polarized cells and at the tight junction level of polarized cells. Thus, the localization of InsP3 receptor in MDCK depends on polarity. PMID- 12759373 TI - Specific inhibition of pathological prion protein accumulation by small interfering RNAs. AB - Development of transmissible spongiform encephalopathies (TSEs) pathogenesis requires the presence of both the normal host prion protein (PrP-sen) and the abnormal pathological proteinase-K resistant isoform (PrP-res). PrP-res forms highly insoluble aggregates, with self-perpetuating properties, by binding and converting PrP-sen molecules into a likeness of themselves. In the present report, we show that small interfering RNA (siRNA) duplexes trigger specific Prnp gene silencing in scrapie-infected neuroblastoma cells. A non-passaged, scrapie infected culture transfected with siRNA duplexes is depleted of PrP-sen and rapidly loses its PrP-res content. The use of different murine-adapted scrapie strains and host cells did not influence the siRNA-induced gene silencing efficiency. More than 80% of transfected cells were positive for the presence of fluorescein-labeled siRNA duplexes. No cytotoxicity associated with the use of siRNA was observed during the time course of these experiments. Despite a transient abrogation of PrP-res accumulation, our results suggest that the use of siRNA may provide a new and promising therapeutic approach against prion diseases. PMID- 12759374 TI - Temporally and spatially selective loss of Rec8 protein from meiotic chromosomes during mammalian meiosis. AB - Sister chromatid cohesion is maintained from DNA replication to metaphase-to anaphase transition by multisubunit protein complexes called cohesin, which include at least four proteins, SMC1alpha, SMC3, Rad21 and either SA1 or SA2, in mammalian somatic cells. We report here the first evidence of the involvement of Rec8 protein, a mammalian homolog of yeast Rec8p, in meiosis-specific chromosome behavior in mammals. In immunoblotting and immunohistochemical analysis using specific antibodies against mouse Rec8, we found that Rec8 was expressed in the testis but not in the kidney or liver; more precisely, it was expressed in spermatocytes and spermatids but not in spermatogonia or other somatic cells. We also found that Rec8 is present in both phosphorylated and dephosphorylated states in vivo. Immunoprecipitation analyses revealed that Rec8 associates with other cohesin proteins, SMC1beta (meiosis-specific protein) and SMC3 and with a component of synaptonemal complexes, SCP3, but not with SMC1alpha. In meiotic chromosome spreads, Rec8 was localized along the axial/lateral elements of the synaptonemal complexes in meiotic prophase from the leptotene to diplotene stages. At later stages, diakinesis and metaphase I, Rec8 was localized along the interstitial axes of chromosomes, including both centromere and arm regions of chromosomes. However, concomitantly with separation of homologous chromosomes in anaphase I, Rec8 was no longer detected along the arm regions, while it persisted on centromere regions up to metaphase II. In anaphase II, the centromeric signals were diminished. We propose from these results that mammalian Rec8 protein, in association with SMC3 and SMC1beta but not SMC1alpha, is involved in meiosis specific chromosome behavior, and that homologous chromosome separation is triggered by selective loss of Rec8 from chromosome arms in meiosis I, while sister chromatid cohesion is maintained until metaphase II/anaphase II transition by centromeric Rec8 during mammalian meiosis. PMID- 12759375 TI - Fission yeast meu14+ is required for proper nuclear division and accurate forespore membrane formation during meiosis II. AB - Using a meiosis-specific subtracted cDNA library of Schizosaccharomyces pombe, we identified meu14+ as a gene whose expression is upregulated during meiosis. Transcription of meu14+ is induced abruptly after the cell enters meiosis. Its transcription is dependent on the meiosis-specific transcription factor Mei4. In meu14Delta cells, the segregation and modification of the SPBs (spindle pole bodies) and microtubule elongation during meiosis II were aberrant. Meiotic meu14Delta cells consequently produced a high frequency of abnormal tetranucleate cells harboring aberrant forespore membranes and failed to produce asci. In wild type cells harboring the integrated meu14+-gfp fusion gene, Meu14-GFP first appeared inside the nuclear region at prophase II, after which it accumulated beside the two SPBs at metaphase II. Thereafter, it formed two ring-shaped structures that surrounded the nucleus at early anaphase II. At post-anaphase II, it disappeared. Meu14-GFP appears to localize at the border of the forespore membrane that later develops into spore walls at the end of sporulation. This was confirmed by coexpressing Spo3-HA, a component of the forespore membrane, with Meu14-GFP. Taken together, we conclude that meu14+ is crucial in meiosis in that it participates in both the nuclear division during meiosis II and the accurate formation of the forespore membrane. PMID- 12759376 TI - Mitofusin-1 protein is a generally expressed mediator of mitochondrial fusion in mammalian cells. AB - Mitochondrial fusion may regulate mitochondrial morphogenesis and underlie complementation between mitochondrial genomes in mammalian cells. The nuclear encoded mitochondrial proteins Mfn1 and Mfn2 are human homologues of the only known protein mediators of mitochondrial fusion, the Drosophila Fzo GTPase and Saccharomyces cerevisiae yFzo1p. Although the Mfn1 and Mfn2 genes were broadly expressed, the two genes showed different levels of mRNA expression in different tissues. Two Mfn1 transcripts were detected at similar levels in a variety of human tissues and were dramatically elevated in heart, while Mfn2 mRNA was abundantly expressed in heart and muscle tissue but present only at low levels in many other tissues. Human Mfn1 protein localized to mitochondria and participated in a high molecular weight, detergent extractable protein complex. Forced expression of Mfn1 in cultured cells caused formation of characteristic networks of mitochondria. Introduction of a point mutation in the conserved G1 region of the predicted GTPase domain (Mfn1K88T) dramatically decreased formation of mitochondrial networks upon Mfn1 overexpression, suggesting that network formation required completion of the Mfn1 GTPase cycle. Conversely, a protein variant carrying a point mutation in the G2 motif of the Mfn1 GTPase domain acted as a dominant negative: overexpression of Mfn1T109A resulted in fragmentation of mitochondria. We propose that Mfn1T109A interferes with fusion activity of endogenous Mfn1 protein, possibly by binding necessary cofactors, so to allow unopposed mitochondrial fission. Thus, Mfn1 appears to be a key player in mediating mitochondrial fusion and morphology in mammalian cells. PMID- 12759377 TI - Taking action on the volume-quality relationship: how long can we hide our heads in the colostomy bag? PMID- 12759378 TI - Potential pitfalls in the use of surrogate endpoints in colorectal adenoma chemoprevention. PMID- 12759379 TI - The volume-outcome relationship: busier hospitals are indeed better, but why? PMID- 12759380 TI - Stat bite: Effect of hospital volume on operative mortality for major cancer surgery. PMID- 12759381 TI - Research council reviews draft fallout report. PMID- 12759383 TI - Researchers suggest that universal 'law' governs tumor growth. PMID- 12759384 TI - National tumor bank set up in United kingdom. PMID- 12759388 TI - Relation of hospital volume to colostomy rates and survival for patients with rectal cancer. AB - BACKGROUND: Postoperative mortality after some types of cancer surgery is inversely related to the number of operations performed at a hospital (i.e., hospital volume). This study assessed the association of hospital volume with colostomy rates and survival for patients with rectal cancer in a large representative cohort identified from the California Cancer Registry. METHODS: We identified 7257 patients diagnosed from January 1, 1994, through December 31, 1997, with stage I-III rectal cancer who underwent surgical resection. Registry data were linked to hospital discharge abstracts and ZIP-code-level data from the 1990 U.S. Census. Associations of hospital volume with permanent colostomy and 30 day mortality were assessed with the Mantel-Haenszel trend test and logistic regression. Overall survival was examined with the Kaplan-Meier method and a multivariable Cox proportional hazards model. Multivariable analyses adjusted for demographic and clinical variables and patient clustering within hospitals. All tests of statistical significance were two-sided. RESULTS: In unadjusted analyses across decreasing quartiles of hospital volume, we observed statistically significant increases in colostomy rates (29.5%, 31.8%, 35.2%, and 36.6%; P<.001) and in 30-day postoperative mortality (1.6%, 1.6%, 2.9%, and 4.8%; P<.001) and a decrease in 2-year survival (83.7%, 83.2%, 80.9%, and 76.6%; P<.001). The adjusted risks of permanent colostomy (odds ratio [OR] = 1.37, 95% confidence interval [CI] = 1.10 to 1.70), 30-day mortality (OR = 2.64, 95% CI = 1.41 to 4.93), and 2-year mortality (hazard ratio = 1.28, 95% CI = 1.15 to 1.44) were greater for patients at hospitals in the lowest volume quartile than for patients at hospitals in the highest volume quartile. Stratification by tumor stage and comorbidity index did not appreciably affect the results. Adjusted colostomy rates varied statistically significantly (P<.001) among individual hospitals independent of volume. CONCLUSIONS: Rectal cancer patients who underwent surgery at high-volume hospitals were less likely to have a permanent colostomy and had better survival rates than those treated in low-volume hospitals. Identifying processes of care that contribute to these differences may improve patients' outcomes in all hospitals. PMID- 12759389 TI - Neoplastic and antineoplastic effects of beta-carotene on colorectal adenoma recurrence: results of a randomized trial. AB - BACKGROUND: In two large, randomized prevention trials, supplementation with beta carotene increased the risk of lung cancer. Subjects in these studies were predominantly cigarette smokers, and the adverse effects were concentrated among those who also drank alcohol. Although beta-carotene supplementation appeared not to increase the risk of cancer generally, it is not clear if smoking and/or alcohol use alters the effect of beta-carotene on carcinogenesis at sites outside the lung. METHODS: We studied the effect of beta-carotene supplementation on colorectal adenoma recurrence among subjects in a multicenter double-blind, placebo-controlled clinical trial of antioxidants for the prevention of colorectal adenomas. A total of 864 subjects who had had an adenoma removed and were polyp-free were randomly assigned (in a factorial design) to receive beta carotene (25 mg or placebo) and/or vitamins C and E in combination (1000 mg and 400 mg, respectively, or placebo), and were followed with colonoscopy for adenoma recurrence 1 year and 4 years after the qualifying endoscopy. A total of 707 subjects had two follow-up examinations and provided smoking and alcohol use data. Adjusted multivariate risk ratios (RRs) and 95% confidence intervals (CIs) were used to assess the effects of beta-carotene on adenoma recurrence. RESULTS: Among subjects who neither smoked cigarettes nor drank alcohol, beta-carotene was associated with a marked decrease in the risk of one or more recurrent adenomas (RR = 0.56, 95% CI = 0.35 to 0.89), but beta-carotene supplementation conferred a modest increase in the risk of recurrence among those who smoked (RR = 1.36, 95% CI = 0.70 to 2.62) or drank (RR = 1.13, 95% CI = 0.89 to 1.43). For participants who smoked cigarettes and also drank more than one alcoholic drink per day, beta carotene doubled the risk of adenoma recurrence (RR = 2.07, 95% CI = 1.39 to 3.08; P for difference from nonsmoker/nondrinker RR <.001). CONCLUSION: Alcohol intake and cigarette smoking appear to modify the effect of beta-carotene supplementation on the risk of colorectal adenoma recurrence. PMID- 12759390 TI - p16/cyclin-dependent kinase inhibitor 2A deficiency in human melanocyte senescence, apoptosis, and immortalization: possible implications for melanoma progression. AB - BACKGROUND: The melanoma susceptibility locus cyclin-dependent kinase inhibitor 2A encodes two unrelated cell growth inhibitors, p16 and alternative reading frame (ARF). In fibroblasts, both proteins are implicated in cellular senescence, a key barrier to tumor development. The p16 coding sequence is more often mutated in melanoma families than is the ARF sequence. To investigate the role of p16 in melanocytes, we assessed aspects of growth, apoptosis, and immortalization in melanocytes cultured from two melanoma patients, both of whom had two inactive p16 alleles but functional ARF. METHODS: Growth and senescence were evaluated by cumulative population-doubling curves, and apoptosis by terminal deoxytransferase labeling. Expression of p53 and p21, which are associated with fibroblast senescence, was assessed by immunoblotting. Amphotropic retroviruses were used to transfer exogenous gene sequences into the melanocytes. RESULTS: Both melanocyte cultures showed high rates of apoptosis, which were reduced when the cells were grown in the presence of keratinocyte feeder cells or human stem cell factor plus endothelin 1. With these growth factors, both cultures proliferated for 45-55 net population doublings, markedly longer than the maximum of 10 net population doublings of normal adult human melanocytes in similar media, indicating impaired senescence. One of the cultures developed chromosomal aberrations, with numerous dicentric chromosomes at senescence, consistent with telomere dysfunction. p53 and p21 levels were not elevated in senescent normal melanocytes but were elevated in senescent p16-deficient melanocytes. Interference with p53 function by transfer of human papillomavirus 16-E6 further extended the lifespan of p16 deficient melanocytes. Human telomerase reverse transcriptase was sufficient to immortalize both these cell strains but not normal melanocytes. CONCLUSION: Normal senescence in human melanocytes requires p16 activity. p53 contributes to a delayed form of senescence that requires telomere shortening, in p16-deficient melanocytes. These findings provide some basis for the role of p16 in melanoma susceptibility. PMID- 12759391 TI - Dual mechanisms for lysophosphatidic acid stimulation of human ovarian carcinoma cells. AB - BACKGROUND: Lysophosphatidic acid (LPA), at concentrations present in ascitic fluid, indirectly stimulates the growth of malignant ovarian tumors by increasing the expression of vascular endothelial growth factor (VEGF) in ovarian cancer cells. We investigated whether LPA could also directly promote ovarian tumor growth by increasing the level of cyclin D1, a key G1-phase checkpoint regulator, which thereby increases cell proliferation. METHODS: Expression of cyclin D1 and LPA receptors (EDG4 and EDG7) was determined in six ovarian cancer cell lines (including OVCAR-3 cells) and immortalized ovarian surface epithelial cells (IOSE 29). Cyclin D1 promoter activity was measured in LPA-treated OVCAR-3 cells cotransfected with cyclin D1 promoter-driven luciferase constructs and cDNA expression plasmids for IkappaBalphaM (a nuclear factor kappaB [NFkappaB] super repressor). RESULTS: Four of six cancer cell lines, including OVCAR-3, overexpressed cyclin D1 protein relative to levels in IOSE-29 cells. LPA treatment increased cyclin D1 protein in a dose- and time-dependent manner in OVCAR-3 cells but not in IOSE-29 cells. LPA stimulated cyclin D1 promoter activity (3.0-fold, 95% confidence interval [CI] = 2.7-fold to 3.3-fold). Mutation of the NFkappaB-binding site in the cyclin D1 promoter to block NFkappaB binding and expression of IkappaBalphaM, which binds NFkappaB and inhibits its binding to the promoter, markedly diminished LPA stimulation of cyclin D1 promoter activity (activity stimulated only 1.4-fold, 95% CI = 1.1-fold to 1.7 fold, and 0.7-fold, 95% CI = 0.6-fold to 0.8-fold, respectively). EDG4 was overexpressed in all cancer cell lines studied relative to that in IOSE-29 cells, but EDG7 was overexpressed in only two lines. CONCLUSIONS: Dual mechanisms are probably involved in LPA stimulation of ovarian tumor growth in vivo. In addition to the previously characterized indirect mechanism that increases angiogenesis via VEGF, LPA may directly increase the level of cyclin D1 in ovarian cancer cells, increasing their proliferation. PMID- 12759392 TI - Identification of a protein fragment of interleukin 2 responsible for vasopermeability. AB - BACKGROUND: The cytokine interleukin 2 (IL-2) is involved in the activation of T cells and has been shown to play a central role in cancer immunotherapy. The full therapeutic potential of IL-2, however, has not been realized because of its dose limiting systemic toxicity. We sought to identify a region of IL-2 that is responsible for the induction of vasopermeability (leaky tumor endothelium), a property associated with the toxicity of the molecule. METHODS: Intact IL-2 or overlapping synthetic peptides of IL-2 that were chemically conjugated to tumor targeting monoclonal antibodies (TNT-1 or Lym-1) were injected into groups of mice (n = 4) that had previously been xenotransplanted with human tumor cells (ME 180 cervical carcinoma and Raji lymphoma). Two hours later, mice received intravenous injections of radiolabeled tracer antibody, and 3 days later they were subjected to biodistribution analysis to measure the ability of each immunoconjugate to enhance tumor uptake of the tracer antibody (i.e., vasopermeability activity). The cytokine activity of the immunoconjugates was determined by assaying their ability to promote the proliferation of a mouse IL-2 dependent cell line. RESULTS: Pretreatment of mice with an antibody/IL-2 immunoconjugate resulted in an approximately fourfold increase in radiolabeled tracer antibody uptake in the xenograft tumor as compared with uptake in mice injected with antibody alone. One synthetic fragment consisting of amino acids 22 58 contained 100% of the vasopermeability activity of IL-2 and was designated permeability-enhancing peptide (PEP). PEP had vasopermeability activity only when conjugated to a tumor-targeting antibody, had maximal activity as a dimer, and was devoid of cytokine activity. CONCLUSIONS: The identification of PEP should aid in the discovery of ways to decrease the toxicity of IL-2. Moreover, PEP is a promising candidate for the generation of agents that can enhance the delivery of antibodies and drugs to tumors. PMID- 12759394 TI - Re: Association of volume and volume-independent factors with accuracy in screening mammogram interpretation. PMID- 12759393 TI - Serum selenium levels in relation to markers of neoplastic progression among persons with Barrett's esophagus. AB - BACKGROUND: Persons with Barrett's esophagus have a substantially greater risk of esophageal adenocarcinoma than the general population. Higher serum selenium levels have been associated with a reduced risk of several cancers; however, their association with the risk of esophageal adenocarcinoma is unknown. We used a cross-sectional study to investigate the relationship between serum selenium levels and markers of neoplastic progression among persons with Barrett's esophagus. METHODS: Medical history, blood, and esophageal tissue specimens were collected from 399 members of a cohort study of Barrett's esophagus patients undergoing endoscopic surveillance. Serum selenium levels were measured by flameless atomic absorption spectrophotometry. DNA content of tissue samples was measured by flow cytometry. Loss of heterozygosity (LOH) at 9p and 17p, chromosomal regions which include the p16 and p53 tumor suppressors, respectively, was detected by automated fluorescent genotyping. Logistic regression was used to calculate odds ratios (ORs) and 95% confidence intervals (CIs). All statistical tests were two-sided. RESULTS: Persons with serum selenium levels in the upper three quartiles (i.e., >1.5 micro M) were less likely to have high-grade dysplasia (OR = 0.5, 95% CI = 0.3 to 0.9) or aneuploidy (OR = 0.4, 95% CI = 0.2 to 0.8) than those with levels in the lowest quartile. Serum selenium levels in the upper three quartiles were associated with similar reductions in risk of 17p (p53) LOH (OR = 0.5, 95% CI = 0.2 to 0.9) and increased 4N fraction (OR = 0.6, 95% CI = 0.3 to 1.2). By contrast, serum selenium levels were not associated with 9p (p16) LOH (OR = 1.0, 95% CI = 0.5 to 1.7), a marker that appears early in neoplastic progression. CONCLUSION: Our preliminary results, from a cross-sectional analysis with biologic markers, suggest that higher serum selenium levels may be associated with a reduced risk of esophageal adenocarcinoma among persons with Barrett's esophagus. Because serum selenium was not associated with 9p (p16) LOH, we speculate that selenium may act primarily at later stages of progression toward adenocarcinoma. PMID- 12759395 TI - Re: Cancer risks in BRCA1 carriers: time for the next generation of studies. PMID- 12759397 TI - Re: Randomized trial of breast irradiation schedules after lumpectomy for women with lymph node-negative breast cancer. PMID- 12759399 TI - Re: Helicobacter pylori and interleukin 1 genotyping: an opportunity to identify high-risk individuals for gastric carcinoma. PMID- 12759400 TI - Re: Double-blind, placebo-controlled, randomized phase III trial of darbepoetin alfa in lung cancer patients receiving chemotherapy. PMID- 12759404 TI - Re: Beta-carotene and lung cancer: a lesson for future chemoprevention investigations? PMID- 12759406 TI - Cutting edge: major CD8 T cell response to live bacillus Calmette-Guerin is mediated by CD1 molecules. AB - MHC class I-restricted CD8(+) T cells are a crucial component of the host defense against mycobacterial infection in mice, but it has often proved very difficult to identify the CD8 T cell response in humans. Human group 1 CD1 molecules (CD1a, -b, -c) mediate MHC-independent presentation of mycobacteria-derived lipid and glycolipid Ags to CD8(+) T cells, and their intracellular localization to the endocytic system may favor efficient monitoring of phagosome-resident mycobacteria. Here, we show that bacillus Calmette-Guerin (BCG)-immunized subjects contain a significant circulating pool of CD8(+) T cells that recognize BCG-infected DCs in a CD1-dependent, but MHC-independent, manner. These CD1 restricted T cells efficiently detected live, rather than dead, BCG and produced IFN-gamma, an important cytokine for protection against mycobacterial infection. These results emphasize that lipid-reactive CD8(+) T cells may contribute to host defense against mycobacterial infection. PMID- 12759407 TI - Cutting edge: bradykinin induces IL-12 production by dendritic cells: a danger signal that drives Th1 polarization. AB - Dendritic cells play a major role in the induction of both innate and acquired immune responses against pathogenic invaders. These cells are also able to sense endogenous activation signals liberated by injured tissues even in the absence of infection. In the present work, we demonstrate that kinins mobilize dendritic cells to produce IL-12 through activation of the B(2) bradykinin receptor subtype and that bradykinin-induced IL-12 responses are tightly regulated both by angiotensin-converting enzyme, a kinin-degrading peptidase, and by endogenous IL 10. Using a mouse model of allergic inflammation, we further show that addition of bradykinin to OVA during immunization results in decreased eosinophil infiltration on Ag challenge. The latter effect was demonstrated to be due to IL 12-driven skewing of Ag-specific T cell responses to a type 1 cytokine profile. Our data thus indicate that kinin peptides can serve as danger signals that trigger dendritic cells to produce IL-12 through activation of B(2) bradykinin receptors. PMID- 12759408 TI - Cutting edge: Monarch-1: a pyrin/nucleotide-binding domain/leucine-rich repeat protein that controls classical and nonclassical MHC class I genes. AB - Proteins containing a limited number of N-terminal motifs followed by nucleotide binding domain and leucine-rich repeat regions are emerging as important regulators for immunity. A search of human genome scaffold databases has identified a large family of known and unknown genes, which we have recently called the CATERPILLER (caspase recruitment domain, transcription enhancer, r(purine)-binding, pyrin, lots of leucine repeats) gene family. This work describes the characterization of a new member, Monarch-1. Monarch-1 has four different splice forms due to the differential splicing of leucine-rich repeat motifs. It is expressed in cells of myeloid-monocytic origin. Affymetrix microarrays and small interfering RNA were used to elucidate the downstream effects of Monarch-1 expression in cells including those of myeloid-monocytic origin. These analyses show that Monarch-1 enhances nonclassical and classical MHC class I expression at the level of the promoter, RNA, and protein expression. PMID- 12759409 TI - Differential regulation of human eosinophil IL-3, IL-5, and GM-CSF receptor alpha chain expression by cytokines: IL-3, IL-5, and GM-CSF down-regulate IL-5 receptor alpha expression with loss of IL-5 responsiveness, but up-regulate IL-3 receptor alpha expression. AB - Our recent data suggested that tissue eosinophils may be relatively insensitive to anti-IL-5 treatment. We examined cross-regulation and functional consequences of modulation of eosinophil cytokine receptor expression by IL-3, IL-5 GM-CSF, and eotaxin. Incubation of eosinophils with IL-3, IL-5, or GM-CSF led to reduced expression of IL-5R alpha, which was sustained for up to 5 days. Eosinophils incubated with IL-5 or IL-3 showed diminished respiratory burst and mitogen activated protein kinase kinase phosphorylation in response to further IL-5 stimulation. In contrast to these findings, eosinophil expression of IL-3R alpha was increased by IL-3, IL-5, and GM-CSF, whereas GM-CSF receptor alpha was down regulated by GM-CSF, but was not affected by IL-3 or IL-5. CCR3 expression was down-regulated by IL-3 and was transiently reduced by IL-5 and GM-CSF, but rapidly returned toward baseline. Eotaxin had no effect on receptor expression for IL-3, IL-5, or GM-CSF. Up-regulation of IL-3R alpha by cytokines was prevented by a phosphoinositol 3-kinase inhibitor, whereas this and other signaling inhibitors had no effect on IL-5R alpha down-regulation. These data suggest dynamic and differential regulation of eosinophil receptors for IL-3, IL 5, and GM-CSF by the cytokine ligands. Since these cytokines are thought to be involved in eosinophil development and mobilization from the bone marrow and are present at sites of allergic inflammation, tissue eosinophils may have reduced IL 5R expression and responsiveness, and this may explain the disappointing effect of anti-IL-5 therapy in reducing airway eosinophilia in asthma. PMID- 12759410 TI - Dendritic cells process and present antigens across a range of maturation states. AB - We isolated dendritic cells (DC) from lymphoid organs of mice bearing a transgene for a membrane-bound form of the model protein hen egg white lysozyme (HEL). DC from the spleen had a lower representation of costimulatory molecules and class II MHC molecules than those isolated from lymph nodes and thymi. Splenic DC were capable of further maturation by in vivo treatment of mice with LPS. The immature DC from spleen processed HEL and displayed the chemically dominant epitope as evidenced by FACS analysis. These immature DC also presented this epitope to CD4(+) T cells. Splenic DC from another transgenic mouse (ML-5) containing serum HEL also showed the ability to process and present Ag despite low levels of circulating HEL. In vitro-derived DC from the bone marrow (bone marrow-derived DC) of mHEL mice also displayed immature to mature features and in both cases displayed HEL peptides as well as SDS-stable MHC class II molecules. Immature bone marrow-derived DC also processed exogenous HEL. We conclude that the DC sets normally found in tissue show a scale of maturation features but even the most immature process and present peptides by MHC class II molecules. PMID- 12759411 TI - Stimulation of primary human endothelial cell proliferation by IFN. AB - The IFN family of cytokines has pleiotropic roles in immunity and development. In this study, we provide evidence that IFN can stimulate the proliferation of primary human endothelial cells. This is in contrast to the growth-suppressive effects of IFN observed on transformed human cells, thereby underscoring the distinctive responses of primary human cells. The growth-stimulatory effect of IFN was determined by an increase in DNA synthesis assessed with [(3)H]thymidine incorporation, an increase in G(2) and M cell cycle phases assessed with flow cytometric analysis, and an increase in cell number. Distinct cell types, including primary human fibroblast and smooth muscle cells, were also growth stimulated by IFN. Neutralizing Abs to IFN were used to demonstrate the growth response was mediated specifically by the IFN cytokine. The signaling pathway of type I IFNs activates STAT1 and STAT2. In primary endothelial cells, we demonstrate that STAT3 and STAT5 are also activated, and these STATs may contribute to cellular proliferation. To evaluate possible effectors of positive growth, DNA microarray analyses were performed to assess gene induction in response to IFN. These results reveal changes in the RNA levels of genes in endothelial cells that encode proteins involved in cellular proliferation. PMID- 12759412 TI - Differential regulation of vitamin D receptor and its ligand in human monocyte derived dendritic cells. AB - The functions of dendritic cells (DCs) are tightly regulated such that protective immune responses are elicited and unwanted immune responses are prevented. 1 alpha 25-dihydroxyvitamin D(3) (1 alpha 25(OH)(2)D(3)) has been identified as a major factor that inhibits the differentiation and maturation of DCs, an effect dependent upon its binding to the nuclear vitamin D receptor (VDR). Physiological control of 1 alpha 25(OH)(2)D(3) levels is critically dependent upon 25 hydroxyvitamin D(3)-1 alpha-hydroxylase (1 alpha OHase), a mitochondrial cytochrome P450 enzyme that catalyzes the conversion of inactive precursor 25 hydroxyvitamin D(3) (25(OH)D(3)) to the active metabolite 1 alpha 25(OH)(2)D(3). Using a human monocyte-derived DC (moDC) model, we have examined the relationship between DC VDR expression and the impact of exposure to its ligand, 1 alpha 25(OH)(2)D(3). We show for the first time that moDCs are able to synthesize 1 alpha 25(OH)(2)D(3) in vitro as a consequence of increased 1 alpha OHase expression. Following terminal differentiation induced by a diverse set of maturation stimuli, there is marked transcriptional up-regulation of 1 alpha OHase leading to increased 1 alpha OHase enzyme activity. Consistent with this finding is the observation that the development and function of moDCs is inhibited at physiological concentrations of the inactive metabolite 25(OH)D(3). In contrast to 1 alpha OHase, VDR expression is down-regulated as monocytes differentiate into immature DCs. Addition of 1 alpha 25(OH)(2)D(3) to moDC cultures at different time points indicates that its inhibitory effects are greater in monocyte precursors than in immature DCs. In conclusion, differential regulation of endogenous 1 alpha 25(OH)(2)D(3) ligand and its nuclear receptor appear to be important regulators of DC biology and represent potential targets for the manipulation of DC function. PMID- 12759413 TI - Generation of cytotoxic responses in mice and human individuals against hematological malignancies using survivin-RNA-transfected dendritic cells. AB - Survivin is a member of the inhibitors of apoptosis family and is overexpressed in many types of human cancers, making it an attractive target for T cell-based immunotherapeutic strategies. Recently, HLA-A2-binding peptides derived from the survivin protein were identified as capable of inducing specific T cell responses in cancer patients. Here we demonstrate that human survivin-specific CTLs generated from PBMC by stimulation with autologous dendritic cells transfected with survivin-RNA were cytotoxic for a range of hemopoietic malignant cell lines and primary tumor cells isolated from patients with acute myeloid leukemia. We also show that vaccination of mice with survivin-RNA-transfected dendritic cells leads to long term resistance to challenge by a survivin-expressing lymphoma, demonstrating the potential of survivin as a tumor rejection Ag. Our data provide evidence for the use of survivin as a target structure for immunotherapeutic strategies against hematological neoplasms. PMID- 12759414 TI - NK cell tolerance in mixed allogeneic chimeras. AB - Alterations in inhibitory receptor expression on NK cells have been detected in mixed allogeneic chimeras and in mosaic MHC class I-expressing transgenic mice. However, it is not known whether or not NK cells are tolerant to host and donor Ags in mixed chimeras. In vitro studies have shown a lack of mutual tolerance of separated donor and host NK cells obtained from mixed chimeras. Using BALB/c-->B6 fully MHC-mismatched mixed chimeras, we have now investigated this question in vivo. Neither donor nor host NK cells in mixed chimeras showed evidence for activation, as indicated by expression of B220 and Thy-1.2 on NK cells in chimeric mice at levels similar to those in nonchimeric control mice. Lethally irradiated, established mixed BALB/c--> B6 chimeras rejected a low dose of beta(2)-microglobulin-deficient bone marrow cells (BMC) efficiently but did not reject BALB/c or B6 BMCs. In contrast, similarly conditioned B6 mice rejected both BALB/c and beta(2)-microglobulin-deficient BMCs. Thus, NK cells were specifically tolerant to the donor and the host in mixed allogeneic chimeras. The similar growth of RMA lymphoma cells in both chimeric and control B6 mice further supports the conclusion that donor BALB/c NK cells are tolerant to B6 Ags in chimeras. Administration of a high dose of exogenous IL-2 could not break NK cell tolerance in chimeric mice, suggesting that NK cell tolerance in chimeras is not due to a lack of activating cytokine. No reduction in the level of expression of the activating receptor Ly-49D, recognizing a donor MHC molecule, was detected among recipient NK cells in mixed chimeras. Thus, the present studies demonstrate that NK cells in mixed chimeras are stably tolerant to both donor and host Ags, by mechanisms that are as yet unexplained. PMID- 12759415 TI - Mature but not immature Fas ligand (CD95L)-transduced human monocyte-derived dendritic cells are protected from Fas-mediated apoptosis and can be used as killer APC. AB - Several in vitro and animal studies have been performed to modulate the interaction of APCs and T cells by Fas (CD95/Apo-1) signaling to delete activated T cells in an Ag-specific manner. However, due to the difficulties in vector generation and low transduction frequencies, similar studies with primary human APC are still lacking. To evaluate whether Fas ligand (FasL/CD95L) expressing killer APC could be generated from primary human APC, monocyte-derived dendritic cells (DC) were transduced using the inducible Cre/Loxp adenovirus vector system. Combined transduction of DC by AdLoxpFasL and AxCANCre, but not single transduction with these vectors, resulted in dose- and time-dependent expression of FasL in >70% of mature DC (mDC), whereas <20% of immature DC (iDC) expressed FasL. In addition, transduction by AdLoxpFasL and AxCANCre induced apoptosis in >80% of iDC, whereas FasL-expressing mDC were protected from FasL/Fas (CD95/Apo 1)-mediated apoptosis despite coexpression of Fas. FasL-expressing mDC eliminated Fas(+) Jurkat T cells as well as activated primary T cells by apoptosis, whereas nonactivated primary T cells were not deleted. Induction of apoptosis in Fas(+) target cells required expression of FasL in DC and cell-to-cell contact between effector and target cell, and was not dependent on soluble FasL. Induction of apoptosis in Fas(+) target cells required expression of FasL in DC, cell-to-cell contact between effector and target cell, and was not dependent on soluble FasL. The present results demonstrate that FasL-expressing killer APC can be generated from human monocyte-derived mDC using adenoviral gene transfer. Our results support the strategy to use killer APCs as immunomodulatory cells for the treatment of autoimmune disease and allograft rejection. PMID- 12759416 TI - Memory phenotype of CD8+ T cells in MHC class Ia-deficient mice. AB - B6.K(b-)D(b-) mice are devoid of class Ia but express normal levels of class Ib molecules. They have low levels of CD8 T cells in both the thymus as well as peripheral T cell compartments. Although the percentage of splenic CD8 alpha alpha T cells is increased in these animals, approximately 90% of CD8 T cells are CD8 alpha beta. In contrast to B6 animals, most of the CD8 T cells from these mice have a memory phenotype (CD44(high)CD122(high) CD62L(low)) including both CD8 alpha beta and CD8 alpha alpha subsets. In the thymus of B6.K(b-)D(b-) animals, there is a decrease in the percentage of SP CD8 T cells, although most are CD44(low), similar to that seen in B6 mice. The spleens from day 1-old B6 and B6.K(b-)D(b-) mice have a relatively high proportion of CD44(high)CD62L(low) CD8 T cells. However, by day 28 most CD8 T cells in B6 mice have a naive phenotype while in B6.K(b-)D(b-) mice the memory phenotype remains. Unlike CD44(high) cells that are found in B6 animals, most CD44(high) cells from B6.K(b-)D(b-) mice do not secrete IFN-gamma rapidly upon activation. The paucity of CD8 T cells in B6.K(b-)D(b-) mice might be due in part to their inability to undergo homeostatic expansion. Consistent with this, we found that CD8 T cells from these animals expand poorly in X-irradiated syngeneic hosts compared with B6 CD8 T cells that respond to class Ia Ags. We examined homeostatic expansion of B6 CD8 T cells in single as well as double class Ia knockout mice and were able to estimate the fraction of cells reactive against class Ia vs class Ib molecules. PMID- 12759418 TI - Genetic dissection of V alpha 14J alpha 18 natural T cell number and function in autoimmune-prone mice. AB - Nonobese diabetic (NOD) mice, a model for type I diabetes (TID), have reduced numbers of invariant V alpha 14J alpha 18 TCR alpha-chain-positive natural T (iNKT) cells that do not release IL-4 in response to in vivo activation through their Ag receptor. The deficit in iNKT cell number and function is implicated in immune dysregulation and the etiology of TID. Therefore, we reasoned that the genetic determinant(s) that controls iNKT cell number and function might lie within Idd (insulin-dependent diabetes susceptibility locus) regions, which are known to contain TID resistance or susceptibility genes. A systematic analysis of iNKT cell number and function in Idd congenic mice revealed that neither iNKT cell number nor their inability to rapidly secrete IL-4 in response to acute in vivo activation by Ag underlies the mechanism of protection from diabetes in Idd congenic mice. Moreover, the regulation of iNKT cell number and function appears to be under the control of several genes. The most notable of these map to the Idd4, Idd5, Idd9.1, and Idd13 regions of the mouse genome. Together these findings provide a clue to the genetic mechanism(s) underlying iNKT cell deficiency in NOD mice. PMID- 12759417 TI - A role for the B7-1/B7-2:CD28/CTLA-4 pathway during negative selection. AB - Although costimulation plays an important role in activating naive T cells, its role in negative selection is controversial. By following thymocyte deletion induced by endogenous superantigens in mice lacking B7-1 and/or B7-2, we have identified a role for both B7-1 and B7-2 in negative selection. Studies using CD28-deficient and CD28/CTLA-4-double-deficient mice have revealed that either CD28 or another as yet undefined coreceptor can mediate these B7-dependent signals that promote negative selection. Finally, CTLA-4 delivers signals that inhibit selection, suggesting that CTLA-4 and CD28 have opposing functions in thymic development. Combined, the data demonstrate that B7-1/B7-2-dependent signals help shape the T cell repertoire. PMID- 12759419 TI - IL-23 production by cosecretion of endogenous p19 and transgenic p40 in keratin 14/p40 transgenic mice: evidence for enhanced cutaneous immunity. AB - p40, the common subunit of the proinflammatory cytokines IL-12 and IL-23, is produced by resident skin cells. Whereas the in vivo effects of IL-12 are well established, little is known about the role of IL-23 in cutaneous immune responses. In this study we show that p40 transgenic (TG) mice constitutively produce IL-23 (p19/p40), but not IL-12 (p35/p40), in basal keratinocytes by cosecretion of TG p40 with endogenous p19. Repeated injections of rIL-23 in littermate (LM) mice result in an inflammatory skin disease similar to that of p40 TG mice, confirming the proinflammatory activity of IL-23. Furthermore, IL-23 secretion by p40 TG keratinocytes induces elevated numbers of Langerhans cells (LC) with a marked up-regulation of costimulatory molecules, indicating advanced maturation of keratin 14 (K14)/p40 LC when compared with LM LC. At the functional level, freshly isolated K14/p40 LC greatly exceeded LC from LM animals in their capacity to stimulate allogeneic T cell proliferation. To assess whether IL-23 regulates cutaneous immune responses in vivo, we used an allogeneic skin transplantation model. Full thickness skin grafts from K14/p40 donors (H-2(q)) transplanted across a MHC class I and class II barrier onto BALB/c (H-2(d)) recipients were rejected in a significantly accelerated fashion (mean survival time: 8.8 days) when compared with skin grafts from non-TG LM (H-2(q)) (mean survival time: 10.7 days, p < 0.01). Based on these results we propose that IL-23 induced changes of LC may be an important mechanism in directing the outcome of cutaneous immune responses. PMID- 12759420 TI - Stimulation of Toll-like receptor 4 by lipopolysaccharide during cellular invasion by live Salmonella typhimurium is a critical but not exclusive event leading to macrophage responses. AB - Invasion of macrophages by salmonellae induces cellular responses, with the bacterial inducers likely to include a number of pathogen-associated molecular patterns. LPS is one of the prime candidates, but its precise role in the process, especially when presented as a component of live infecting bacteria, is unclear. We thus investigated this question using the lipid A antagonist E5531, the macrophage-like cell line RAW 264.7, and primary macrophage cultures from C3H/HeJ and Toll-like receptor 4(-/-) (TLR-4(-/-)) mice. We show that LPS presented on live salmonellae provides an essential signal, via functional TLR-4, for macrophages to produce NO and TNF-alpha. Furthermore, the mitogen-activated protein kinase c-Jun N-terminal kinase and p38 are activated, and the transcription factor NF-kappa B is translocated to the nucleus when RAW 264.7 cells are presented with purified LPS or live salmonellae. Purified LPS stimulates rapid, transitory mitogen-activated protein kinase activation that is inhibited by E5531, whereas bacterial invasion stimulates delayed, prolonged activation, unaffected by E5531. Both purified LPS and bacterial invasion caused translocation of NF-kappa B, but whereas E5531 always inhibited activation by purified LPS, activation by bacterial invasion was only inhibited at later time points. In conclusion, we show for the first time that production of NO and TNF alpha is critically dependent on activation of TLR-4 by LPS during invasion of macrophages by salmonellae, but that different patterns of activation of intracellular signaling pathways are induced by purified LPS vs live salmonellae. PMID- 12759421 TI - TCR signal transduction in antigen-specific memory CD8 T cells. AB - Memory T cells are more responsive to Ag than naive cells. To determine whether memory T cells also have more efficient TCR signaling, we compared naive, effector, and memory CD8 T cells of the same antigenic specificity. Surprisingly, initial CD3 signaling events are indistinguishable. However, memory T cells have more extensive lipid rafts with higher phosphoprotein content before TCR engagement. Upon activation in vivo, they more efficiently induce phosphorylation of-LAT (linker for activation of T cells), ERK (extracellular signal-regulated kinase), JNK (c-Jun N-terminal kinase), and p38. Thus, memory CD8 T cells do not increase their TCR sensitivity, but are better poised to augment downstream signals. We propose that this regulatory mechanism might increase signal transduction in memory T cells, while limiting TCR cross-reactivity and autoimmunity. PMID- 12759422 TI - IL-21 in synergy with IL-15 or IL-18 enhances IFN-gamma production in human NK and T cells. AB - NK and T cell-derived IFN-gamma is a key cytokine that stimulates innate immune responses and directs adaptive T cell response toward Th1 type. IL-15, IL-18, and IL-21 have significant roles as activators of NK and T cell functions. We have previously shown that IL-15 and IL-21 induce the expression of IFN-gamma, T-bet, IL-12R beta 2, and IL-18R genes both in NK and T cells. Now we have studied the effect of IL-15, IL-18, and IL-21 on IFN-gamma gene expression in more detail in human NK and T cells. IL-15 clearly activated IFN-gamma mRNA expression and protein production in both cell types. IL-18 and IL-21 enhanced IL-15-induced IFN gamma gene expression. IL-18 or IL-21 alone induced a modest expression of the IFN-gamma gene but a combination of IL-21 and IL-18 efficiently up-regulated IFN gamma production. We also show that IL-15 activated the binding of STAT1, STAT3, STAT4, and STAT5 to the regulatory sites of the IFN-gamma gene. Similarly, IL-21 induced the binding of STAT1, STAT3, and STAT4 to these elements. IL-15- and IL 21-induced STAT1 and STAT4 activation was verified by immunoprecipitation with anti-phosphotyrosine Abs followed by Western blotting with anti-STAT1 and anti STAT4 Abs. IL-18 was not able to induce the binding of STATs to IFN-gamma gene regulatory sites. IL-18, however, activated the binding of NF-kappa B to the IFN gamma promoter NF-kappa B site. Our results suggest that both IL-15 and IL-21 have an important role in activating the NK cell-associated innate immune response. PMID- 12759423 TI - Propagation and control of T cell responses by heparan sulfate-bound IL-2. AB - IL-2, a cytokine produced by T cells, is a key regulator of immune responses and T cell homeostasis. Controlling the availability of IL-2 is consequently of significant import to the immune system. Like other cytokines, IL-2 is thought to function as a soluble agonist, transiently present when secreted in response to appropriate stimuli. In this study, we show that the most salient properties of IL-2, propagation and control of T cell responses, are mediated in vivo by bound and not free cytokine and specifically by heparan sulfate-bound IL-2. These findings necessitate a new look at how IL-2 regulates immune responses and support the notion that the microenvironment plays a determining role in modulating the character of immune responses. PMID- 12759424 TI - Isolated lymphoid follicle formation is inducible and dependent upon lymphotoxin sufficient B lymphocytes, lymphotoxin beta receptor, and TNF receptor I function. AB - The gastrointestinal mucosa contains a complex network of lymphoid compartments that have evolved to efficiently protect the host from invading pathogens. Recently, an additional lymphoid structure resembling Peyer's patches (PP) in composition and architecture has been identified in the murine small intestine, the isolated lymphoid follicle (ILF). In this study we examine the nature and factors required for ILF formation. We observed a spectrum of structures fitting the previous descriptions of ILFs, ranging from clusters of B220(+) cells (which we have termed immature ILFs) to well-organized lymphoid nodules (which we have termed mature ILFs). Here we demonstrate that that similar to PP formation, ILF formation requires lymphotoxin (LT)- and LT beta receptor-dependent events. However unlike PP formation, the LT- and LT beta receptor-dependent events required for ILF formation can occur in adulthood and require LT-sufficient B lymphocytes. We demonstrate that mature ILF formation occurs in response to lumenal stimuli, including normal bacterial flora, and requires TNF receptor I function. These findings suggest that ILFs are organized intestinal lymphoid structures whose formation can be induced and whose mass can be expanded in response to mucosal challenges. PMID- 12759425 TI - Drug-inducible, dendritic cell-based genetic immunization. AB - Determining the mechanism of Ag loading of Langerhans cells (LC) for genetic immunization (GI) is complicated by the inability to distinguish between the response generated by direct transfection of LC from that due to exogenous uptake. To unravel this mechanism, we examined the impact of gene gun treatment on LC with respect to their activation and migration from skin, transgene expression, and ability to initiate humoral and cellular immune responses upon transfer to naive mice. To assess responses generated by direct LC transfection, an RU486-inducible expression system was used as a GI vector. In vitro skin organ cultures were developed from gene gun immunized mouse ear specimens to obtain LC. Gene gun treatment markedly augmented (3-fold) LC migration from ear skin, and these LC expressed the transgene at RNA and protein levels. Transfer of 2 x 10(5) migratory cells resulted in identical cellular responses to, but 10-fold lower humoral responses than, standard GI. Using an RU486-inducible system, we were able to measure responses generated by directly transfected LC. Our results indicate that direct transfection is a predominant pathway for LC Ag loading. The ability to regulate transgene expression with inducible DC-based vaccines demonstrates a new level of immunological control. PMID- 12759426 TI - IL-12 administration accelerates autoimmune diabetes in both wild-type and IFN gamma-deficient nonobese diabetic mice, revealing pathogenic and protective effects of IL-12-induced IFN-gamma. AB - IL-12 administration to nonobese diabetic (NOD) mice induces IFN-gamma-secreting type 1 T cells and high circulating IFN-gamma levels and accelerates insulin dependent diabetes mellitus (IDDM). Here we show that IL-12-induced IFN-gamma production is dispensable for diabetes acceleration, because exogenous IL-12 could enhance IDDM development in IFN-gamma-deficient as well as in IFN-gamma sufficient NOD mice. Both in IFN-gamma(+/-) and IFN-gamma(-/-) NOD mice, IL-12 administration generates a massive and destructive insulitis characterized by T cells, macrophages, and CD11c(+) dendritic cells, and increases the number of pancreatic CD4(+) cells secreting IL-2 and TNF-alpha. Surprisingly, IL-12-induced IFN-gamma hinders pancreatic B cell infiltration and inhibits the capacity of APCs to activate T cells. Although pancreatic CD4(+) T cells from IL-12-treated IFN-gamma(-/-) mice fail to up-regulate the P-selectin ligand, suggesting that their entry into the pancreas may be impaired, T cell expansion is favored in these mice compared with IL-12-treated IFN-gamma(+/-) mice because IL-12 administration in the absence of IFN-gamma leads to enhanced cell proliferation and reduced T cell apoptosis. NO, an effector molecule in beta cell destruction, is produced ex vivo in high quantity by pancreas-infiltrating cells through a mechanism involving IL-12-induced IFN-gamma. Conversely, in IL-12-treated IFN gamma-deficient mice, other pathways of beta cell death appear to be increased, as indicated by the up-regulated expression of Fas ligand on Th1 cells in the absence of IFN-gamma. These data demonstrate that IFN-gamma has a dual role, pathogenic and protective, in IDDM development, and its deletion allows IL-12 to establish alternative pathways leading to diabetes acceleration. PMID- 12759427 TI - Lung CD25 CD4 regulatory T cells suppress type 2 immune responses but not bronchial hyperreactivity. AB - To study the effects of chronic Ag deposition in the airway mucosa on CD4(+) T cell priming and subsequent airway disease, transgenic mice were generated that expressed OVA under the control of the surfactant protein C promoter. CD4 T cells from these mice were tolerant to OVA but this was overcome among spleen CD4 T cells by crossing to OVA-specific DO11.10 TCR-transgenic mice. Lungs from the double-transgenic mice developed lymphocytic infiltrates and modest mucus cell hyperplasia. Infiltrating cells were unaffected by the absence of either Rag-1 or Stat6, although the latter deficiency led to the disappearance of mucus. In the lung of double-transgenic mice, a large number of Ag-specific CD4 T cells expressed CD25 and functioned as regulatory T cells. The CD25(+) CD4 T cells suppressed proliferation of CD25(-) CD4 T cells in vitro and inhibited type 2 immune responses induced by aerosolized Ags in vivo. Despite their ability to suppress allergic type 2 immunity in the airways, however, CD25(+) CD4 regulatory T cells had no effect on the development of bronchial hyperreactivity. PMID- 12759428 TI - Selective induction of dendritic cells using granulocyte macrophage-colony stimulating factor, but not fms-like tyrosine kinase receptor 3-ligand, activates thyroglobulin-specific CD4+/CD25+ T cells and suppresses experimental autoimmune thyroiditis. AB - Fms-like tyrosine kinase receptor 3-ligand (Flt3-L) and GM-CSF cause expansion of different subsets of dendritic cells and skew the immune response toward predominantly Th1 and Th2 type, respectively. In the present study, we investigated their effects on experimental autoimmune thyroiditis in CBA/J mice. Relative to mouse thyroglobulin (mTg) immunized controls, mTg-immunized mice treated with Flt3-L showed more severe thyroiditis characterized by enhanced lymphocytic infiltration of the thyroid, and IFN-gamma and IL-2 production. In contrast, mice treated with GM-CSF, either before or after immunization with mTg, showed suppressed T cell response to mTg and failed to develop thyroiditis. Lymphocytes from these mice, upon activation with mTg in vitro, produced higher levels of IL-4 and IL-10. Additionally, GM-CSF-treated mice showed an increase in the frequency of CD4(+)/CD25(+) T cells, which suppressed the mTg-specific T cell response. Neutralization of IL-10, but not IL-4, or depletion of CD4(+)/CD25(+) cells resulted in increased mTg-specific in vitro T cell proliferation suggesting that IL-10 produced by the Ag-specific CD4(+)/CD25(+) regulatory T cells might be critical for disease suppression. These results indicate that skewing immune response toward Th2, through selective activation of dendritic cells using GM CSF, may have therapeutic potential in Th1 dominant autoimmune diseases including Hashimoto's thyroiditis. PMID- 12759429 TI - Thyroid-specific expression of IFN-gamma limits experimental autoimmune thyroiditis by suppressing lymphocyte activation in cervical lymph nodes. AB - The role of IFN-gamma in the pathogenesis of autoimmune disease is controversial, being described as immunostimulatory in some studies and immunosuppressive in others. To determine the contribution of local expression of IFN-gamma, we derived NOD.H-2(h4) transgenic mice overexpressing IFN-gamma in a thyroid restricted manner. Transgenic mice, which had serum IFN-gamma levels similar to wild-type littermates, showed up-regulation of MHC class II on thyrocytes, but did not develop spontaneous thyroiditis. Upon immunization with murine thyroglobulin, transgenic mice developed milder disease and reduced IgG1 responses compared with wild type. The milder disease was associated with decreased frequency of activated CD44(+) lymphocytes in the cervical lymph nodes. This suppressive effect was confirmed by showing that blockade of systemic IFN gamma with mAb enhanced disease and increased IgG1 responses. The study supports a disease-limiting role of IFN-gamma in autoimmune thyroiditis. Furthermore, it provides the first evidence that local IFN-gamma activity in the thyroid is sufficient for disease suppression. PMID- 12759430 TI - Absence of allograft ICAM-1 attenuates alloantigen-specific T cell priming, but not primed T cell trafficking into the graft, to mediate acute rejection. AB - The expression and function of ICAM-1 are critical components in the initiation and elicitation of many T cell-mediated responses. Whether ICAM-1 expression is required on the T cells or on the APC during T cell priming remains unclear. To address this issue in alloantigen-specific T cell activation, the priming and function of T cells in response to heart allografts from MHC-mismatched wild-type vs ICAM-1(-/-) donors were tested. Wild-type C57BL/6 (H-2(b)) heart allografts were rejected by A/J (H-2(a)) recipients on days 7-9, whereas B6.ICAM-1(-/-) allografts survived until days 18-23 post-transplant. On day 7 post-transplant, infiltrating macrophages and CD4(+) and CD8(+) T cells in the ICAM-1(-/-) allografts were 20-30% those observed in the wild-type allografts. ELISPOT analyses indicated that the number of alloantigen-specific T cells producing IFN gamma from recipients of ICAM-1-deficient grafts was 60% lower than that from recipients of wild-type allografts. On day 16 post-transplant, these numbers did not markedly increase in ICAM-1-deficient allograft recipients. Consistent with the reduced priming of alloreactive T cells, isolated dendritic cells from ICAM 1(-/-) mice stimulated allogeneic T cell proliferation poorly compared with wild type dendritic cells. When A/J mice were primed with wild-type dendritic cells and then received wild-type or ICAM-1-deficient heart allografts 3 days later, the primed recipients rejected the wild-type and ICAM-1(-/-) allografts on days 5 6 post-transplant. These results indicate that optimal priming of alloreactive T cells requires allograft expression of ICAM-1, but, once primed, recipient T cell infiltration into the allograft is independent of graft ICAM-1 expression. PMID- 12759431 TI - Notch signaling induces multilineage myeloid differentiation and up-regulates PU.1 expression. AB - Hemopoietic commitment is initiated by and depends on activation of transcription factors. However, it is unclear whether activation of lineage-affiliated transcription factors is extrinsically regulated by to date unknown agents or is the result of a cell autonomous program. Here we show that signaling by the Notch1 transmembrane receptor instructively induces myeloid differentiation of multipotent hemopoietic progenitor cells and concomitantly up-regulates the expression of the transcription factor PU.1. Transient activation of Notch1 signaling is sufficient to irreversibly reduce self-renewal of multipotent progenitor cells accompanied by increased and accelerated differentiation along the granulocyte, macrophage, and dendritic cell lineages. Activated Notch1 has no direct influence on apoptosis of multipotent progenitor cells, shows a weak inhibition of proliferation, and does not substitute for survival and proliferation signals provided by cytokines. Activated Notch1 directly increases PU.1 RNA levels, leading to a high concentration of PU.1 protein, which has been shown to direct myeloid differentiation. These findings identify Notch as an extrinsic regulator of myeloid commitment, and the lineage-affiliated transcription factor PU.1 as a specific direct target gene of Notch. PMID- 12759432 TI - Yin Yang 1 is a lipopolysaccharide-inducible activator of the murine 3' Igh enhancer, hs3. AB - The 3' Igh enhancers, DNase I hypersensitive site (hs) 3B and/or hs4, are required for germline transcription, and hence, class switch recombination for multiple isotypes. A number of hs3-binding transcription factors have been identified by EMSA, including octamer and NF-kappa B family members, and Pax5. We have found that the binding of the transcription factor, Yin Yang 1 (YY1), to hs3 and to the mu E1 site of the intronic enhancer, E mu, is induced in primary splenic B cells after approximately 48 h in response to LPS and other activators of class switch recombination. Transient transfection experiments in B cell lines indicate that YY1 is an activator of hs3. Interestingly, levels of YY1 expression are unchanged in resting and LPS-stimulated B cells. Mixing experiments followed by EMSA showed that a protein present in resting B cells prevented binding of YY1 to DNA. We found that recombinant retinoblastoma protein (Rb) inhibited binding of YY1 to hs3 in a dose-dependent manner, and we have identified complexes of endogenous YY1 with the Rb in resting B cells, but not in LPS-stimulated B cells. A difference in Rb phosphorylation state was also confirmed between resting (G(0)) B cells and LPS-stimulated B cells. These observations suggest that the interaction of YY1 with hypophosphorylated Rb in resting B cells prevents interaction of YY1 with DNA. After stimulation with class-switching activators, such as LPS, Rb becomes hyperphosphorylated and YY1 is released and can then bind to the hs3 enhancer and E mu. PMID- 12759433 TI - Normal somatic hypermutation of Ig genes in the absence of 8-hydroxyguanine-DNA glycosylase. AB - The hypermutation cascade in Ig V genes can be initiated by deamination of cytosine in DNA to uracil by activation-induced cytosine deaminase and its removal by uracil-DNA glycosylase. To determine whether damage to guanine also contributes to hypermutation, we examined the glycosylase that removes oxidized guanine from DNA, 8-hydroxyguanine-DNA glycosylase (OGG1). OGG1 has been reported to be overexpressed in human B cells from germinal centers, where mutation occurs, and could be involved in initiating Ab diversity by removing modified guanines. In this study, mice deficient in Ogg1 were immunized, and V genes from the H and kappa L chain loci were sequenced. Both the frequency of mutation and the spectra of nucleotide substitutions were similar in ogg1(-/-) and Ogg1(+/+) clones. More importantly, there was no significant increase in G:C to T:A transversions in the ogg1(-/-) clones, which would be expected if 8 hydroxyguanine remained in the DNA. Furthermore, Ogg1 was not up-regulated in murine B cells from germinal centers. These findings show that hypermutation is unaffected in the absence of Ogg1 activity and indicate that 8-hydroxyguanine lesions most likely do not cause V gene mutations. PMID- 12759434 TI - Uncoupling of promitogenic and antiapoptotic functions of IL-2 by Smad-dependent TGF-beta signaling. AB - TGF-beta opposes proliferative signaling by IL-2 through mechanisms that remain incompletely defined. In a well-characterized CD8(+) T cell model using wild-type and mutated IL-2 receptors, we examined the effects of TGF-beta on distinct IL-2 signaling events in CD8(+) T cells. IL-2 induces c-myc, cyclin D2, and cyclin E in a redundant manner through the Shc and STAT5 pathways. TGF-beta inhibited the ability of either the Shc or STAT5 pathway to induce these genes, as well as T cell proliferation. The inhibitory effects of TGF-beta were reversed by expression of a dominant-negative form of Smad3. TGF-beta did not impair proximal signaling by Shc or STAT5, and induction of some downstream genes, including cytokine-inducible Src homology-2-containing protein (CIS), bcl-x(L), and bcl-2, was spared. Experiments with c-fos, cyclin D2, and CIS reporter genes revealed that promoter-proximal regulatory elements dictate the sensitivity of IL-2 target genes to inhibition by TGF-beta. By leaving the Shc and STAT5 pathways functional while inhibiting their target genes selectively, TGF-beta was found to uncouple the proliferative and antiapoptotic functions of IL-2. Thus, TGF-beta is not a simple antagonist of IL-2, but rather serves to qualitatively modify the IL-2 signal to create a unique pattern of gene expression that neither cytokine can induce independently. PMID- 12759435 TI - IL-18 receptor beta-induced changes in the presentation of IL-18 binding sites affect ligand binding and signal transduction. AB - IL-18 is a pleiotropic proinflammatory cytokine that is involved in induction of inflammatory mediators, regulation of the cytotoxic activity of NK cells and T cells, and differentiation and activation of both Th1 and Th2 cells. IL-18 signals through its specific cell surface receptor IL-18R, which comprises two subunits: IL-18R alpha and IL-18R beta. IL-18R alpha alone has a weak affinity for IL-18 binding, while the IL-18R alpha/beta complex has a high affinity. By using several anti-IL-18 mAbs and IL-18 binding protein, we have examined whether these site-specific inhibitors could block the binding of IL-18 to IL-18R alpha and to the IL-18R alpha/beta complex. Here we show that IL-18 binding to IL-18R alpha was inhibited by a neutralizing mAb, 125-2H, while binding of IL-18 to the alpha/beta receptor complex was not. This suggests that IL-18R beta-induced conformational changes may occur in IL-18R alpha upon dimerization, leading to changes in the presentation of IL-18 binding sites. Epitope mapping of 125-2H using human-mouse IL-18 chimeras identified a region in IL-18 that was required for 125-2H recognition. This region, as examined by IL-18R binding and functional analysis, appeared to be critical for triggering signal transduction through the heterodimeric receptor. PMID- 12759436 TI - Novel variants of the IL-10 receptor 1 affect inhibition of monocyte TNF-alpha production. AB - IL-10-deficient mice exhibit spontaneous enterocolitis and other symptoms akin to Crohn's disease, indicating that IL-10 might regulate normal physiology in the gut. However, clinical trials with IL-10 in Crohn's disease were disappointing, although some patients showed healing of intestinal mucosa. This study searched for genetic polymorphisms within the IL-10 pathway. We decided to screen for mutations of the IL-10R1 cDNA in healthy volunteers and Crohn's disease patients and identified two novel variants: a serine 138-to-glycine (S138G) and a glycine 330-to-arginine (G330R) substitution. The allelic frequency in a European cohort was relatively high (16% for the S138G and 33% for the G330R), and S138G was in strong linkage disequilibrium with G330R. A similar allele frequency was found in a group of Crohn's patients. In IL-10R1 G330R-expressing monocytes, the inhibitory effect of IL-10 on TNF-alpha production was diminished, indicating that this variant may be a loss-of-function allele. No such difference was observed between haplotypes 4 (G330R only) and 7 (S138G and G330R). In addition, these IL-10R1 variants had no influence on the IL-10R1 expression density. Structural analysis of the S138G variant revealed that the substitution of S138G may interfere with binding of IL-10 to IL-10R1. PMID- 12759437 TI - Wound healing and expression of antimicrobial peptides/polypeptides in human keratinocytes, a consequence of common growth factors. AB - In addition to acting as a physical barrier against microorganisms, the skin produces antimicrobial peptides and proteins. After wounding, growth factors are produced to stimulate the regeneration of tissue. The growth factor response ceases after regeneration of the tissue, when the physical barrier protecting against microbial infections is re-established. We found that the growth factors important in wound healing, insulin-like growth factor I and TGF-alpha, induce the expression of the antimicrobial peptides/polypeptides human cationic antimicrobial protein hCAP-18/LL-37, human beta-defensin 3, neutrophil gelatinase associated lipocalin, and secretory leukocyte protease inhibitor in human keratinocytes. Both an individual and a synergistic effect of these growth factors were observed. These findings offer an explanation for the expression of these peptides/polypeptides in the skin disease psoriasis and in wound healing and define a host defense role for growth factors in wound healing. PMID- 12759438 TI - Mast cell activation by Mycobacterium tuberculosis: mediator release and role of CD48. AB - Mast cells (MC) are abundant in the lung and other peripheral tissue, where they participate in inflammatory processes against bacterial infections. Like other effector cells of the innate immune system, MC interact directly with a wide variety of infectious agents. This interaction results in MC activation and inflammatory mediator release. We demonstrated that MC interact with Mycobacterium tuberculosis, triggering the release of several prestored reagents, such as histamine and beta-hexosaminidase, and de novo synthesized cytokines, such as TNF-alpha and IL-6. A number of M. tuberculosis Ags, ESAT-6, MTSA-10, and MPT-63, have been implicated in MC activation and mediator release. A MC plasmalemmal protein, CD48, was implicated in interactions with mycobacteria because CD48 appeared to aggregate in the MC membrane at sites of bacterial binding and because Abs to CD48 inhibited the MC histamine response to mycobacteria. Cumulatively, these findings suggest that MC, even in the absence of opsonins, can directly recognize M. tuberculosis and its Ags and have the potential to play an active role in mediating the host's innate response to M. tuberculosis infection. PMID- 12759439 TI - Renewal of peripheral CD8+ memory T cells during secondary viral infection of antibody-sufficient mice. AB - Kinetic studies and short pulses of injected 5-bromo-2-deoxyuridine have been used to analyze the development and renewal of peripheral CD8(+) memory T cells in the lungs during primary and secondary respiratory virus infections. We show that developing peripheral CD8(+) memory T cells proliferate during acute viral infection with kinetics that are indistinguishable from those of lymphoid CD8(+) memory T cells. Secondary exposure to the same virus induces a new round of T cell proliferation and extensive renewal of the peripheral and lymphoid CD8(+) memory T cell pools in both B cell-deficient mice and mice with immune Abs. In mice with virus-specific Abs, CD8(+) T cell proliferation takes place with minimal inflammation or effector cell recruitment to the lungs. The delayed arrival of CD8(+) memory T cells to the lungs of these animals suggests that developing memory cells do not require the same inflammatory signals as effector cells to reach the lung airways. These studies provide important new insight into mechanisms that control the maintenance and renewal of peripheral memory T cell populations during natural infections. PMID- 12759441 TI - Hepatitis C virus core and nonstructural protein 3 proteins induce pro- and anti inflammatory cytokines and inhibit dendritic cell differentiation. AB - Antiviral immunity requires recognition of viral pathogens and activation of cytotoxic and Th cells by innate immune cells. In this study, we demonstrate that hepatitis C virus (HCV) core and nonstructural protein 3 (NS3), but not envelope 2 proteins (E2), activate monocytes and myeloid dendritic cells (DCs) and partially reproduce abnormalities found in chronic HCV infection. HCV core or NS3 (not E2) triggered inflammatory cytokine mRNA and TNF-alpha production in monocytes. Degradation of I-kappa B alpha suggested involvement of NF-kappa B activation. HCV core and NS3 induced production of the anti-inflammatory cytokine, IL-10. Both monocyte TNF-alpha and IL-10 levels were higher upon HCV core and NS3 protein stimulation in HCV-infected patients than in normals. HCV core and NS3 (not E2) inhibited differentiation and allostimulatory capacity of immature DCs similar to defects in HCV infection. This was associated with elevated IL-10 and decreased IL-2 levels during T cell proliferation. Increased IL-10 was produced by HCV patients' DCs and by core- or NS3-treated normal DCs, while IL-12 was decreased only in HCV DCs. Addition of anti-IL-10 Ab, not IL-12, ameliorated T cell proliferation with HCV core- or NS3-treated DCs. Reduced allostimulatory capacity in HCV core- and NS3-treated immature DCs, but not in DCs of HCV patients, was reversed by LPS maturation, suggesting more complex DC defects in vivo than those mediated by core or NS3 proteins. Our results reveal that HCV core and NS3 proteins activate monocytes and inhibit DC differentiation in the absence of the intact virus and mediate some of the immunoinhibitory effects of HCV via IL-10 induction. PMID- 12759440 TI - Subversion and utilization of the host cell cyclic adenosine 5' monophosphate/protein kinase A pathway by Brucella during macrophage infection. AB - Brucella spp. are intramacrophage pathogens that induce chronic infections in a wide range of mammals, including domestic animals and humans. Therefore, the macrophage response to infection has important consequences for both the survival of phagocytosed bacteria and the further development of host immunity. However, very little is known about the macrophage cell signaling pathways initiated upon infection and the virulence strategy that Brucella use to counteract these responses and secure their survival. In a previous study, we have shown that macrophages activated by SR141716A, a ligand of the cannabinoid receptor CB1, acquired the capacity to control Brucella and observed that the CB1 receptor triggering engages the microbicidal activity of phagocytes. To analyze the perturbation of cell signaling pathway during macrophage infection by Brucella, we hypothesized that SR141716A provides cell signaling that interferes with the bacterial message leading to inhibition of macrophage functions. As CB1 receptor belongs to the family of G protein-linked receptors, we explored the cAMP signaling pathway. In this study, we show that the CB1 ligand inhibited the bacteria-induced cell signaling. Taking advantage of this result, we then demonstrated that Brucella infection elicited a rapid activation of the cAMP/protein kinase A pathway. This activation resulted in a prolonged phosphorylation of the transcription factor CREB. We finally demonstrate that the activation of the cAMP/protein kinase A pathway is crucial for the survival and establishment of Brucella within macrophages. For the first time in phagocytes, we thus characterized a primordial virulence strategy of Brucella involving the host signaling pathway, a novel point of immune intervention of this virulent pathogen. PMID- 12759442 TI - Dendritic cell-based immunotherapy combined with antimony-based chemotherapy cures established murine visceral leishmaniasis. AB - Dendritic cells (DCs) have been proposed to play a critical role as adjuvants in vaccination and immunotherapy. In this study we evaluated the combined effect of soluble Leishmania donovani Ag (SLDA)-pulsed syngeneic bone marrow-derived DC based immunotherapy and antimony-based chemotherapy for the treatment of established murine visceral leishmaniasis. Three weekly injections of SLDA-pulsed DCs into L. donovani-infected mice reduced liver and splenic parasite burden significantly, but could not clear parasite load from these organs completely. Strikingly, the conventional antileishmanial chemotherapy (sodium antimony gluconate) along with injections of SLDA-pulsed DCs resulted in complete clearance of parasites from both these organs. Repetitive in vitro stimulation of splenocytes from uninfected or L. donovani-infected mice with SLDA-pulsed DCs led to the emergence of CD4(+) T cells with characteristics of Th1 cells. Our data indicate that DC-based immunotherapy enhances the in vivo antileishmanial potential of antimony or vice versa. PMID- 12759443 TI - IKK beta plays an essential role in the phosphorylation of RelA/p65 on serine 536 induced by lipopolysaccharide. AB - Activation of the I kappa B kinase (IKK) complex by LPS induces phosphorylation and degradation of I kappa B alpha, leading to the nuclear translocation of NF kappa B. Although it is essential for NF-kappa B activation, emerging evidence has indicated that the nuclear translocation of NF-kappa B is not sufficient to activate NF-kappa B-dependent transcription. Here, we reported that LPS induced the phosphorylation of the p65 trans-activation domain on serine 536 in monocytes/macrophages. Using mouse embryonic fibroblasts lacking either IKK alpha or IKK beta, we found that IKK beta played an essential role in LPS-induced p65 phosphorylation on serine 536, while IKK alpha was partially required for the p65 phosphorylation. The LPS-induced p65 phosphorylation on serine 536 was independent of the phosphatidylinositol 3'-kinase/Akt signaling pathway. Furthermore, we found that the phosphorylation on serine 536 increased the p65 transcription activity. In summary, our results demonstrate that IKK beta plays an essential role in the LPS-induced p65 phosphorylation on serine 536, which may represent a mechanism to regulate the NF-kappa B transcription activity by LPS. PMID- 12759444 TI - Effective mucosal immunity to anthrax: neutralizing antibodies and Th cell responses following nasal immunization with protective antigen. AB - Mucosal, but not parenteral, immunization induces immune responses in both systemic and secretory immune compartments. Thus, despite the reports that Abs to the protective Ag of anthrax (PA) have both anti-toxin and anti-spore activities, a vaccine administered parenterally, such as the aluminum-adsorbed anthrax vaccine, will most likely not induce the needed mucosal immunity to efficiently protect the initial site of infection with inhaled anthrax spores. We therefore took a nasal anthrax vaccine approach to attempt to induce protective immunity both at mucosal surfaces and in the peripheral immune compartment. Mice nasally immunized with recombinant PA (rPA) and cholera toxin (CT) as mucosal adjuvant developed high plasma PA-specific IgG Ab responses. Plasma IgA Abs as well as secretory IgA anti-PA Abs in saliva, nasal washes, and fecal extracts were also induced when a higher dose of rPA was used. The anti-PA IgG subclass responses to nasal rPA plus CT consisted of IgG1 and IgG2b Abs. A more balanced profile of IgG subclasses with IgG1, IgG2a, and IgG2b Abs was seen when rPA was given with a CpG oligodeoxynucleotide as adjuvant, suggesting a role for the adjuvants in the nasal rPA-induced immunity. The PA-specific CD4(+) T cells from mice nasally immunized with rPA and CT as adjuvant secreted low levels of CD4(+) Th1-type cytokines in vitro, but exhibited elevated IL-4, IL-5, IL-6, and IL-10 responses. The functional significance of the anti-PA Ab responses was established in an in vitro macrophage toxicity assay in which both plasma and mucosal secretions neutralized the lethal effects of Bacillus anthracis toxin. PMID- 12759446 TI - Rac1 deletion in mouse neutrophils has selective effects on neutrophil functions. AB - Defects in myeloid cell function in Rac2 knockout mice underline the importance of this isoform in activation of NADPH oxidase and cell motility. However, the specific role of Rac1 in neutrophil function has been difficult to assess since deletion of Rac1 results in embryonic lethality in mice. To elucidate the specific role of Rac1 in neutrophils, we generated mice with a conditional Rac1 deficiency restricted to cells of the granulocyte/monocyte lineage. As observed in Rac2-deficient neutrophils, Rac1-deficient neutrophils demonstrated profound defects in inflammatory recruitment in vivo, migration to chemotactic stimuli, and chemoattractant-mediated actin assembly. In contrast, superoxide production is normal in Rac1-deficient neutrophils but markedly diminished in Rac2 null cells. These data demonstrate that although Rac1 and Rac2 are both required for actin-mediated functions, Rac2 is specifically required for activation of the neutrophil NADPH oxidase. PMID- 12759445 TI - Urokinase-type plasminogen activator potentiates lipopolysaccharide-induced neutrophil activation. AB - Urokinase plasminogen activator (uPA) is a serine protease that catalyzes the conversion of plasminogen to plasmin. Although increased circulating levels of uPA are present in endotoxemia and sepsis, conditions in which activated neutrophils contribute to the development of acute organ dysfunction, the ability of uPA to participate directly in LPS-induced neutrophil activation has not been examined. In the present experiments, we show that uPA can enhance activation of neutrophils exposed to submaximal stimulatory doses of LPS. In particular, uPA increased LPS-induced activation of intracellular signaling pathways, including Akt and c-Jun N-terminal kinase, nuclear translocation of the transcriptional regulatory factor NF-kappa B, and expression of proinflammatory cytokines, including IL-1 beta, macrophage-inflammatory protein-2, and TNF-alpha. There was no effect of uPA on LPS-induced activation of p38 mitogen-activated protein kinase in neutrophils. Transgenic mice unable to produce uPA (uPA(-/-)) were protected from endotoxemia-induced lung injury, as determined by development of lung edema, pulmonary neutrophil accumulation, lung IL-1 beta, macrophage inflammatory protein-2, and TNF-alpha cytokine levels. These results demonstrate that uPA can potentiate LPS-induced neutrophil responses and also suggest that such effects are sufficiently important in vivo to play a major contributory role in neutrophil-mediated inflammatory responses, such as the development of acute lung injury. PMID- 12759447 TI - CC chemokine ligand 5/RANTES chemokine antagonists aggravate glomerulonephritis despite reduction of glomerular leukocyte infiltration. AB - The chemokine CC chemokine ligand (CCL)5/RANTES as well as its respective receptor CCR5 mediate leukocyte infiltration during inflammation and are up regulated early during the course of glomerulonephritis (GN). We tested the effects of the two CCL5/RANTES blocking analogs, Met-RANTES and amino-oxypentane RANTES, on the course of horse apoferritin (HAF)-induced GN. HAF-injected control mice had proliferative GN with mesangial immune complex deposits of IgG and HAF. Daily i.p. injections of Met-RANTES or amino-oxypentane-RANTES markedly reduced glomerular cell proliferation and glomerular macrophage infiltration, which is usually associated with less glomerular injury and proteinuria in HAF-GN. Surprisingly, however, HAF-GN mice treated with both analogs showed worse disease with mesangiolysis, capillary obstruction, and nephrotic range albuminuria. These findings were associated with an enhancing effect of the CCL5/RANTES analogs on the macrophage activation state, characterized by a distinct morphology and increased inducible NO synthetase expression in vitro and in vivo, but a reduced uptake of apoptotic cells in vivo. The humoral response and the Th1/Th2 balance in HAF-GN and mesangial cell proliferation in vitro were not affected by the CCL5/RANTES analogs. We conclude that, despite blocking local leukocyte recruitment, chemokine analogs can aggravate some specific disease models, most likely due to interactions with systemic immune reactions, including the removal of apoptotic cells and inducible NO synthetase expression. PMID- 12759448 TI - Tryptase precursors are preferentially and spontaneously released, whereas mature tryptase is retained by HMC-1 cells, Mono-Mac-6 cells, and human skin-derived mast cells. AB - Tryptase (alpha and beta) levels in serum are used to assess mast cell involvement in human disease. Using cultured cells, the current study examines the hypothesis that protryptase(s) are spontaneously secreted by mast cells at rest, whereas mature tryptase(s) are stored in secretory granules until their release by activated cells. HMC-1 cells have only beta-tryptase genes and the corresponding mRNA. Mono-Mac-6 cells have both alpha- and beta-tryptase genes but preferentially express alpha-tryptase. Mono-Mac-6 cells spontaneously secrete most of their tryptase, which consists of alpha-protryptase, whereas mature tryptase is retained inside these cells. HMC-1 cells also spontaneously secrete most of their tryptase, identified as beta-protryptase, and retain mature tryptase. Skin-derived mast cells retain most of their tryptase, which is mature, and spontaneously secrete protryptase(s). Total tryptase levels in plasma are detectable but no different in healthy subjects with and without the gene for alpha-tryptase, consistent with pro forms of both alpha- and beta-tryptase being spontaneously secreted. Thus, protryptase(s) are spontaneously secreted by resting mast cells, whereas mature tryptase is retained by mast cells until they are activated to degranulate. PMID- 12759449 TI - IFN regulatory factor-1 is required for the up-regulation of the CD40-NF-kappa B activator 1 axis during airway inflammation. AB - Recent studies show that NF-kappa B activator 1 (Act1) functions as an important adapter molecule for CD40-mediated signaling in epithelial cells. To explore the physiological function of the CD40-Act1 axis, we studied the regulation of gene expression of CD40 and Act1 both in vivo and in cell culture models. Although CD40 and Act1 are up-regulated in mouse lung upon LPS stimulation, IL-1 plus IFN alpha, -beta, or -gamma synergistically up-regulate both CD40 and Act1 gene expression in human epithelial A549 cells. Cycloheximide superinduces the Act1 mRNA, whereas actinomycin D completely abolishes the Act1 mRNA, indicating that the induction of Act1 mRNA is at the transcriptional level and does not require protein synthesis. Promoter sequence analyses identified putative IFN regulatory factor (IRF)-1, C/EBP-beta, and AP-1 transcription factor binding sites in the Act1 promoter. Although mutation of any of the three sites abolished the promoter activity, Abs against IRF-1 and C/EBP-beta, but not AP-1, blocked the formation of the DNA-binding complex induced by IL-1 plus IFN-beta stimulation, suggesting cooperative action between IRF-1 and C/EBP-beta in mediating Act1 promoter activity. Importantly, LPS-induced gene expression of CD40 and Act1 in the mouse lung is abolished in IRF-1(-/-) mice, indicating an essential role of transcription factor IRF-1 in the coordinated regulation of these two genes during airway inflammation. The induced expression of the CD40-Act1 axis by inflammatory cytokines in epithelial cells probably plays an important role in priming these cells for their response to CD40 ligand during airway inflammation. PMID- 12759450 TI - Regulation of airway epithelial cell NF-kappa B-dependent gene expression by protein kinase C delta. AB - Airway epithelial cells synthesize proinflammatory molecules such as IL-8, GM CSF, RANTES, and ICAM-1, the expression of which is increased in the airways of patients with asthma. We investigated the regulation of these NF-kappa B dependent genes by the novel protein kinase C (PKC) isoform PKC delta in 16HBE14o human airway epithelial cells, focusing on IL-8 expression. Transient transfection with the constitutively active catalytic subunit of PKC delta (PKC delta-CAT), and treatment with bryostatin 1, an activator of PKC delta, each increased transcription from the IL-8 promoter, whereas overexpression of PKC epsilon had minor effects. Expression of a dominant negative PKC delta mutant (PKC delta-KR) or pretreatment of cells with rottlerin, a chemical PKC delta inhibitor, attenuated TNF-alpha- and phorbol ester-induced transcription from the IL-8 promoter. Bryostatin 1 treatment increased IL-8 protein abundance in primary airway epithelial cells. Selective activation of PKC delta by bryostatin also activated NF-kappa B, as evidenced by p65 RelA and p50 NF-kappa B1 binding to DNA, NF-kappa B trans-activation, and I kappa B degradation. The sufficiency of PKC delta to induce NF-kappa B nuclear translocation and binding to DNA was confirmed in a 16HBE14o- cell line inducibly expressing PKC delta-CAT under the tet-off system. Deletion of the NF-kappa B response element severely attenuated PKC delta-induced IL-8 promoter activity. Finally, PKC delta-CAT induced transcription from the GM-CSF, RANTES, and ICAM-1 promoters. Together these data suggest that PKC delta plays a key role in the regulation of airway epithelial cell NF-kappa B-dependent gene expression. PMID- 12759451 TI - Neutrophil serine proteinases activate human nonepithelial cells to produce inflammatory cytokines through protease-activated receptor 2. AB - Protease-activated receptors (PARs) compose a family of G protein-coupled receptors activated by proteolysis with exposure of their tethered ligand. Recently, we reported that a neutrophil-derived serine proteinase, proteinase 3 (PR3), activated human oral epithelial cells through PAR-2. The present study examined whether other neutrophil serine proteinases, human leukocyte elastase (HLE), and cathepsin G (Cat G) activate nonepithelial cells, human gingival fibroblasts (HGF). HLE and Cat G as well as PR3 activated HGF to produce IL-8 and monocyte chemoattractant protein 1. Human oral epithelial cells but not HGF express mRNA and protein of secretory leukocyte protease inhibitor, an inhibitor of HLE and Cat G, and recombinant secretory leukocyte protease inhibitor clearly inhibited the activation of HGF induced by HLE and Cat G but not by PR3. HGF express PAR-1 and PAR-2 mRNA in the cells and the proteins on the cell surface. HLE and Cat G cleaved the peptide corresponding to the N terminus of PAR-2 with exposure of its tethered ligand. Treatment with trypsin, an agonist for PAR-2, and a synthetic PAR-2 agonist peptide induced intracellular Ca(2+) mobilization and rendered cells refractory to subsequent stimulation with HLE and Cat G. The production of cytokine induced by HLE and Cat G and the PAR-2 agonist peptide was completely abolished by inhibition of phospholipase C. These findings suggest that neutrophil serine proteinases have equal ability to activate human nonepithelial cells through PAR-2 to produce inflammatory cytokines and may control a number of inflammatory processes such as periodontitis. PMID- 12759452 TI - IL-1 alpha, innate immunity, and skin carcinogenesis: the effect of constitutive expression of IL-1 alpha in epidermis on chemical carcinogenesis. AB - Tumor promoters such as the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) are proinflammatory agents, and their mechanism of action in epithelial carcinogenesis has been linked to the release of IL-1 alpha and the induction of chronic inflammation in skin. To test the role of IL-1 alpha and inflammation in models of cutaneous carcinogenesis, we used our previously described FVB/N transgenic mice overexpressing 17-kDa IL-1 alpha in the epidermis under the keratin 14 (K14) promoter. Strikingly, the K14/IL-1 alpha mice were completely resistant to papilloma and carcinoma formation induced by a two-stage DMBA/TPA protocol, while littermate controls developed both tumor types. K14/IL-1 alpha mice crossed with the highly sensitive TG.AC mice, constitutively expressing mutant Ha-Ras, also failed to develop papillomas or carcinomas. When the K14/IL-1 alpha transgene was bred onto a recombinase-activating gene-2-deficient background, the resistance persisted, indicating that innate, but not acquired, mechanisms may be involved in the resistance to the initiation/promotion model. As an alternative approach, a complete carcinogenesis protocol using repetitive application of DMBA alone was applied. Surprisingly, although the IL-1 alpha mice still did not develop papillomas, they did develop carcinomas de novo at an accelerated rate compared with controls. We conclude that constitutive IL-1 alpha expression rendered FVB mice completely resistant to carcinomas that required evolution from prior papillomas, but facilitated carcinomas that did not evolve from papillomas, as in the complete carcinogenesis protocol. Thus, the role of IL 1 alpha and, by extension that of other proinflammatory factors, in epithelial carcinogenesis are more complex than previously appreciated. These mice may provide a mechanism to investigate the validity of these models of human skin tumorigenesis. PMID- 12759453 TI - Synergistic effect of TNF-alpha in soluble VCAM-1-induced angiogenesis through alpha 4 integrins. AB - In our present study we focused on soluble VCAM-1 (sVCAM-1)/alpha(4) integrin induced angiogenesis and found that this type of angiogenesis was mediated through p38 mitogen-activated protein kinase and focal adhesion kinase (FAK). HUVEC expressed both alpha(4) and beta(1) integrins, and it was reported that expression of alpha(4) integrin and its counterreceptor, sVCAM-1/VCAM-1, was enhanced in response to an inflammatory cytokine, TNF-alpha. In endothelial cells phosphorylation of p38 and FAK, but not that of extracellular signal-regulated kinase 1/2 was induced by sVCAM-1. Migration of endothelial cells was stimulated in response to sVCAM-1 at similar levels as those induced by vascular endothelial growth factor, and sVCAM-1-induced migration was almost completely blocked by neutralizing Ab against alpha(4) integrin, by either an inhibitor of p38 (SB203580), or by adenovirus containing FAK-related nonkinase. sVCAM-1 also induced the formation of blood vessels in Matrigel plug assay in vivo, and this neovascularization was blocked by SB203580 or neutralizing Ab against alpha(4) integrin. Moreover, we also confirmed that both TNF-alpha and sVCAM-1 could synergistically induce angiogenesis in the corneas of mice when each factor at used dose could not induce. This angiogenesis by TNF-alpha and sVCAM-1 was almost completely blocked by coadministration of SB203580 and also by neutralizing Ab against alpha(4) integrin. These results suggest that sVCAM-1/alpha(4) integrin induces angiogenesis through p38 and FAK signaling pathways. PMID- 12759455 TI - Localization of MHC class II/human cartilage glycoprotein-39 complexes in synovia of rheumatoid arthritis patients using complex-specific monoclonal antibodies. AB - Recently human cartilage gp-39 (HC gp-39) was identified as a candidate autoantigen in rheumatoid arthritis (RA). To further investigate the relevance of this Ag in RA, we have generated a set of five mAbs to a combination epitope of complexes of HC gp-39(263-275) and the RA-associated DR alpha beta 1*0401 HLA class II molecules. FACS studies revealed that these mAb recognize specific complexes on homozygous DR alpha beta 1*0401-positive B lymphoblastoid cells pulsed with HC gp-39(263-275). The best mAb, 12A, was further characterized using a set of irrelevant DR alpha beta 1*0401-binding peptides and truncated/elongated versions of HC gp-39(263-275) itself. The minimal epitope recognized in combination with DR alpha beta 1*0401 was HC gp-39(263-273). Peptides not encompassing HC gp-39(263-273) were not recognized. Three of five mAb were able to inhibit (up to 90%) the response of HC gp-39(263-275)-specific DR alpha beta 1*0401-restricted T cell hybridomas to peptide-pulsed APC or purified complexes. Using mAb 12A, we have been able to identify and localize dendritic cells that present DR alpha beta 1*0401/HC gp-39(263-275) complexes in synovial tissue of DR alpha beta 1*0401-positive RA patients, indicating local presentation of the HC gp-39(263-275) epitope in the inflamed target tissue by professional APC. These data support a role of HC gp-39 in the local autoimmune response that leads to chronic inflammation and joint destruction. PMID- 12759454 TI - T cells of atopic asthmatics preferentially infiltrate into human bronchial xenografts in SCID mice. AB - T cells play an important role in the pathogenesis of bronchial asthma. However, it is not completely known how circulating lymphocytes infiltrate into the airways of asthmatic patients. Because SCID mice are unable to reject xenogenic transplants, many xenotransplant models using various human tissues have been developed. Therefore, to examine the interaction between bronchi and T lymphocytes of asthma, it may be possible to use the human bronchial xenograft and PBMC xenograft in SCID mice. We transplanted human bronchi into the subcutaneum of SCID mice and i.p. injected PBMCs that were obtained from patients with atopic asthma, atopic dermatitis and rheumatoid arthritis, and normal subjects (asthmatic, dermatitis, rheumatic, and normal huPBMC-SCID mice). There was no difference in the percentage of CD3-, CD4-, CD8-, CD25-, CD45RO-, CD103-, and cutaneous lymphocyte Ag-positive cells in PBMCs among the patients with asthma, dermatitis, rheumatoid arthritis, and normal subjects, and CD3-positive cells in peripheral blood of asthmatic, dermatitis, rheumatic, and normal huPBMC SCID mice. The number of CD3-, CD4-, and CD8-positive cells in the xenografts of asthmatic huPBMC-SCID mice was higher than those of dermatitis, rheumatic, and normal huPBMC-SCID mice. IL-4 mRNA and IL-5 mRNA were significantly higher in the xenografts of asthmatic huPBMC-SCID mice than those in the xenografts of normal huPBMC-SCID mice, but there were no significant differences in the expressions of IL-2 mRNA or IFN-gamma mRNA between them. These findings suggest that T cells, especially Th2-type T cells, of asthmatics preferentially infiltrate into the human bronchi. PMID- 12759456 TI - P2X7 receptor-dependent blebbing and the activation of Rho-effector kinases, caspases, and IL-1 beta release. AB - In response to ATP binding, the P2X7R facilitates cation channel activation, nonspecific pore formation, rapid changes in plasma membrane morphology, and secretion of IL-1 beta from LPS-primed macrophages. To investigate the relationship between the P2X7R-dependent changes in plasma membrane organization and the release of IL-1 beta, we generated time-lapse movies of ATP-stimulated BAC1 murine macrophages in conjunction with biochemical analyses of IL-1 beta release. Similar image analyses in human embryonic kidney 293 cells expressing recombinant P2X7R (HEK-P2X7) permitted comparison of P2X7R-dependent effects in macrophage vs nonmacrophage backgrounds. Whereas HEK-P2X7 cells exhibit zeiotic blebbing within 5 min of ATP treatment, BAC1 macrophages initiated a distinct "tethered" blebbing 10 min after ATP addition. This blebbing was comparably induced by the P2X7R-selective agonist BzATP and was blocked by P2X7R inhibitors KN-62 and oxidized ATP. Blebbing was initiated at ATP concentrations > or = 3 mM, but optimal IL-1 beta release occurred at 1 mM ATP. P2X7R-dependent blebbing was abrogated in the presence of Rho-effector kinase inhibitors Fasudil and Y-27632, but ATP-induced IL-1 beta release was unaffected. ATP-induced activation of RhoA could be detected in both HEK-P2X7 cells and BAC1 murine macrophages. Thus, P2X7R activation signals distinct, novel membrane blebbing events (dependent on RhoA activation and Rho-effector kinase activity) and simultaneously initiates release of IL-1 beta. Our observations that blebbing and IL-1 beta release are dissociable suggest these events occur via parallel rather than convergent signaling pathways. PMID- 12759457 TI - Regulation of lipopolysaccharide sensitivity by IFN regulatory factor-2. AB - IFN regulatory factors (IRFs) are a family of transcription factors and include several members that regulate expression of pro- and anti-inflammatory genes. Mice with a targeted mutation in IRF-2 (IRF-2(-/-)) were studied after injection of LPS to evaluate the importance of IRF-2 in the regulation of endotoxicity. IRF 2(-/-) mice were highly refractory to LPS-induced lethality. Although hepatic TNF alpha mRNA and circulating TNF-alpha were significantly elevated in LPS challenged IRF-2(-/-) mice, levels of IL-1, IL-12, and IFN-gamma mRNA and protein, as well as IL-6 protein, were significantly lower than levels seen in LPS-challenged IRF-2(+/+) mice. IRF-2(-/-) mice were also more refractory to TNF alpha challenge than were control mice, which was consistent with their diminished sensitivity to LPS, yet no significant difference in the mRNA expression of TNFRs was observed. IL-12R beta 2 mRNA levels from LPS-challenged IRF-2(-/-) mice were significantly different after 1, 6, and 8 h, suggesting that both diminished IL-12 and altered IL-12R expression contribute to the paucity of IFN-gamma produced. IRF-2 knockout mice also failed to sustain LPS-inducible levels of IRF-1 and IFN consensus sequence binding protein mRNA expression, two transacting factors required for IL-12 transcription, perhaps as a result of diminished IL-1 beta, IL-6, and IFN-gamma levels. Liver sections from IRF-2(+/+) and IRF-2(-/-) mice were analyzed 6 h after a typically lethal injection of LPS. IRF-2(-/-) mice exhibited greater numbers of apoptotic Kupffer cells than did wild-type mice, suggesting a novel anti-apoptotic role for IRF-2. Collectively, these findings reveal a critical role for IRF-2 in endotoxicity, and point to a previously unappreciated role for IRF-2 in the regulation of apoptosis. PMID- 12759459 TI - Marked airway eosinophilia prevents development of airway hyper-responsiveness during an allergic response in IL-5 transgenic mice. AB - Tissue eosinophilia probably plays important roles in the pathophysiology of bronchial asthma and allergic disorders; however, this concept was challenged recently by controversial results in mouse models of bronchial asthma treated with anti-IL-5 Ab and the failure of anti-IL-5 therapy in humans. We have now used a unique model, IL-5 transgenic (TG) mice, to address a fundamental question: is airway eosinophilia beneficial or detrimental in the allergic response? After sensitization and challenge with OVA, IL-5 TG mice showed a marked airway eosinophilia. Surprisingly, these IL-5 TG mice showed lower airway reactivity to methacholine. Immunohistochemical analysis of the lungs revealed a marked peribronchial infiltration of eosinophils, but no eosinophil degranulation. In vitro, mouse eosinophils from peritoneal lavage fluids did not produce superoxide anion, but did produce an anti-inflammatory and fibrotic cytokine, TGF-beta 1. Indeed, the TGF-beta 1 levels in bronchoalveolar lavage fluid specimens from IL-5 TG mice directly correlated with airway eosinophilia (r = 0.755). Furthermore, anti-IL-5 treatment of IL-5 TG mice decreased both airway eosinophilia and TGF-beta 1 levels in bronchoalveolar lavage fluids and increased airway reactivity. Thus, in mice, marked eosinophilia prevents the development of airway hyper-reactivity during an allergic response. Overall, the roles of eosinophils in asthma and in animal models need to be addressed carefully for their potentially detrimental and beneficial effects. PMID- 12759460 TI - Complement C5a receptor-mediated signaling may be involved in neurodegeneration in Alzheimer's disease. AB - In our earlier results, we demonstrated that cells expressing the complement C5aR are vulnerable since abnormal activation of C5aR caused apoptosis of these cells. In this study, we demonstrate that activation of C5aR by antisense homology box (AHB) peptides synthesized in multiple antigenic peptide form and representing putative interaction sites of the C5a/C5aR evoked calcium influx in TGW neuroblastoma cells. Dose-dependent inhibition of the response was found when the cells were pretreated with C5a, suggesting that C5aR was involved in this process. In addition, pretreatment with monomeric forms of the AHB peptides resulted in attenuation of the calcium signals, supporting the idea of the role of C5aR in this process. Cells of a neuron-rich primary culture and pyramidal cells of rat brain slices also responded to the AHB peptide activation with an increase in the intracellular calcium level, showing that calcium metabolism might be affected in these cells. TUNEL staining demonstrated that C5aR-mediated apoptosis could be induced both in cells of the primary culture as well as in cortical pyramidal neurons of the rat brain. In addition, we investigated expression of C5aR in the hippocampal and cortical neurons of human brains of healthy and demented patients using two anti-human C5aR Abs. Pyramidal cells of the hippocampus and cortex and granular cells of the hippocampus were immunopositive on staining. Although staining was also positive in the vascular dementia brain, it disappeared in the brain with Alzheimer's disease. These results provide further support that C5aR may be involved in neurodegeneration. PMID- 12759458 TI - Expression of a novel murine type I IFN in the pancreatic islets induces diabetes in mice. AB - IFN-kappa belongs to a recently identified subclass of type I IFNs. In this study, we report the cloning and preliminary characterization of the murine homologue of IFN-kappa. The gene encodes a 200-aa protein which is 38.5% homologous to human IFN-kappa. Murine IFN-kappa contains four cysteines in analogous positions to those observed in the IFN-alpha and an additional fifth unique cysteine, C174. The murine gene is located on chromosome 4, where other type I murine IFN genes, IFN-alpha and IFN-beta, are clustered. This region is syntenic with human chromosome 9 where the gene encoding IFN-kappa and the type I IFN gene cluster are found. Mouse IFN-kappa is expressed at low levels in peritoneal macrophages and its expression is up-regulated by dsRNA and IFN-gamma. Similar to previously reported transgenic mice carrying type I and type II IFNs, transgenic mice overexpressing murine IFN-kappa in the beta cells of the pancreas develop overt diabetes with hyperglycemia. Histological characterization of pancreatic islets from these transgenic mice showed inflammatory infiltrates with corresponding destruction of beta cells. PMID- 12759461 TI - Impaired secretion of IL-10 by T cells from patients with common variable immunodeficiency--involvement of protein kinase A type I. AB - Common variable immunodeficiency (CVID) is a heterogeneous group of B cell deficiency syndromes. T cell abnormalities are present in a high proportion of patients with CVID, suggesting impaired T cell-mediated stimulation of B cells. Based on the importance of IL-10 for B cell function and the involvement of the cAMP/protein kinase A type I (PKAI) system in IL-10 synthesis, we examined IL-10 secretion in T cells from CVID patients and controls, particularly focusing on possible modulatory effects of the cAMP/PKAI system. Our main findings were: 1) anti-CD3 and anti-CD3/anti-CD28 activated T cells from CVID patients secreted less IL-10 than healthy controls. This defect was not related to varying proportions of T cell subsets (e.g., CD4(+)/CD8(+), CD45RA(+)/RO(+), or CD28(-) T cells); 2) PKAI activation through the cAMP agonist 8-CPT-cAMP markedly inhibited IL-10 secretion from T cells through CD3 and CD28 activation in both patients and controls, but the sensitivity for cAMP-dependent inhibition was increased in CVID; 3) selective PKAI inhibition by Rp-8-Br-cAMPS markedly increased IL-10 secretion in anti-CD3 and anti-CD3/anti-CD28-stimulated T cells in both patients and controls. Even at the lowest concentrations of Rp-8-Br-cAMPS, IL-10 secretion in CVID patients reached levels comparable to those in controls. Our findings suggest impaired secretion of IL-10 by T cells from CVID patients, suggesting a possible link between T cell deficiency and impaired B cell function in CVID. The involvement of the cAMP/PKAI system in this defect suggests a novel target for therapeutic immunomodulation in CVID. PMID- 12759462 TI - Allospecific T cell epitope sharing reveals extensive conservation of the antigenic features of peptide ligands among HLA-B27 subtypes differentially associated with spondyloarthritis. AB - HLA-B*2702, B*2704, and B*2705 are strongly associated with spondyloarthritis, whereas B*2706 is not. Subtypes differ among each other by a few amino acid changes and bind overlapping peptide repertoires. In this study we asked whether differential subtype association with disease is related to differentially bound peptides or to altered antigenicity of shared ligands. Alloreactive CTL raised against B*2704 were analyzed for cross-reaction with B*2705, B*2702, B*2706, and mutants mimicking subtype changes. These CTL are directed against many alloantigen-bound peptides and can be used to analyze the antigenicity of HLA-B27 ligands on different subtypes. Cross-reaction of anti-B*2704 CTL with B*2705 and B*2702 correlated with overlap of their peptidic anchor motifs, suggesting that many shared ligands have similar antigenic features on these three subtypes. Moreover, the percent of anti-B*2704 CTL cross-reacting with B*2706 was only slightly lower than the overlap between the corresponding peptide repertoires, suggesting that most shared ligands have similar antigenic features on these two subtypes. Cross-reaction with B*2705 or mutants mimicking changes between B*2704 and B*2705 was donor-dependent. In contrast, cross-reaction with B*2702 or B*2706 was less variable among individuals. Conservation of antigenic properties among subtypes has implications for allorecognition, as it suggests that shared peptides may determine cross-reaction across exposed amino acid differences in the MHC molecules and that the antigenic distinctness of closely related allotypes may differ among donors. Our results also suggest that differential association of HLA-B27 subtypes with spondyloarthritis is more likely related to differentially bound peptides than to altered antigenicity of shared ligands. PMID- 12759463 TI - HIV-1 replication increases HIV-specific CD4+ T cell frequencies but limits proliferative capacity in chronically infected children. AB - This study investigated the relationship between HIV-1 replication and virus (HIV 1; CMV)-specific CD4(+) T cell frequency and function in HIV-1-infected children. HIV-1 gag p55-specific CD4(+) T cell IFN-gamma responses were detected in the majority of children studied. p55-specific responses were detected less commonly and at lower frequencies in children with <50 copies/ml plasma HIV-1 RNA than in children with active HIV-1 replication. In children with <50 copies/ml plasma HIV 1, p55-specific responses were detected only in children with evidence of ongoing HIV-1 replication, indicating a direct relationship between HIV-1 replication and HIV-specific CD4(+) T cell frequencies. In contrast, p55-specific proliferative responses were detected more frequently in children with <50 copies/ml plasma HIV 1. CMV-specific CD4(+) responses were more commonly detected and at higher frequencies in CMV-coinfected children with suppressed HIV-1 replication. The lack of HIV-specific CD4(+) proliferative responses, along with the preservation of CMV-specific CD4(+) responses in children with controlled HIV-1 replication, suggests that viral replication may have deleterious effects on HIV-1 and other virus-specific CD4(+) responses. Vaccination to stimulate HIV-specific CD4(+) T cell responses in these children may synergize with antiretroviral therapy to improve the long-term control of viral replication, and may perhaps allow the eventual discontinuation of antiretroviral therapy. PMID- 12759464 TI - Retinoic acid reduces autoimmune renal injury and increases survival in NZB/W F1 mice. AB - Retinoic acids, a group of natural and synthetic vitamin A derivatives, have potent antiproliferative and anti-inflammatory properties. Recently, retinoic acids were reported to inhibit Th1 cytokine production. We investigated the effects of retinoic acid on lupus nephritis in a model of NZB/NZW F(1) (NZB/W F(1)) mice. Three-month-old NZB/W F(1) mice were separated into two groups: one treated with all-trans-retinoic acid (ATRA; 0.5 mg i.p., three times weekly for 7 mo) and one with saline as a control. Compared with controls, ATRA-treated mice survived longer and exhibited a significant reduction of proteinuria, renal pathological findings including glomerular IgG deposits, and serum anti-DNA Abs. Splenomegaly was less marked in the treated mice than in controls. Transcripts encoding IFN-gamma, IL-2, and IL-10 in splenic CD4(+) T cells were significantly reduced in treated mice compared with controls. We conclude that treatment with ATRA in SLE-prone NZB/W F(1) mice significantly alleviates autoimmune renal disorder and prolongs survival; this may thus represent a novel approach to the treatment of patients with lupus nephritis. PMID- 12759465 TI - Validity of the Nottingham Health Profile in a Finnish out-patient population with rheumatoid arthritis. AB - OBJECTIVE: To assess the quality of life of 122 rheumatoid arthritis (RA) patients using the Nottingham Health Profile (NHP), which is a generic quality-of life instrument, and to compare the results with those of 99 healthy local control subjects. METHODS: Our patient group represented consecutive out-patients from a central hospital. The NHP contains 38 items, each with a weighted score, that assess subjective distress on six dimensions: mobility, pain, energy, sleep, emotional reactions and social isolation. RESULTS: The median (age-adjusted) NHP scores for mobility, pain and energy showed statistically highly significant differences (P < 0.001) between the RA patients and the control group, indicating a poorer quality of life among the RA patients on all these dimensions. These NHP dimensions were also closely related to patients' experience of their overall health status. The NHP scores for sleep, emotional reaction and social isolation did not differ between the patients and the controls. CONCLUSION: The NHP, an instrument for assessing health-related quality of life, differentiated RA patients from local healthy individuals on the dimensions of mobility, energy and pain. PMID- 12759466 TI - A national qualitative survey of community-based musculoskeletal services in the UK. AB - OBJECTIVE: To determine the characteristics of community-based musculoskeletal services provided by primary care organizations within the UK. METHODS: Members of five professional groups within UK primary care organizations (n=461) were sent a questionnaire. RESULTS: The response rate by organization was 71% (328/461). Respondents described 350 community-based musculoskeletal services, 233/328 (71%) organizations had one or more musculoskeletal services within their community. Five main forms of service provision were: a scanning service, a rehabilitation service, physiotherapy services, joint or soft tissue injections and the implementation of integrated care pathways. In 162 services, patients were assessed, reviewed or triaged, by a 'non-consultant' health-care professional (physiotherapist or general practitioner), at an intermediate level between primary and secondary care. The purpose of the service was described in 292/350 services, only 39/350 had clear evaluation strategies and 53/350 had consideration of individual training needs. CONCLUSIONS: There are a wide range of musculoskeletal services flourishing within the community sector, whose quality may be variable. Whilst there is good evidence to suggest systematic planning of these services, we are concerned about the lack of data to support their effectiveness in terms of clinical outcomes, monitoring of service delivery standards and ongoing professional development of service providers. Commissioners of such services must ensure they have taken account of the evidence base and met any identified needs of local populations. Services should have a clear purpose with an appropriate evaluation strategy, and well-defined dissemination mechanisms. An integrated educational strategy for staff within the service must be sensitive to issues relating to accreditation, appraisal and revalidation. PMID- 12759467 TI - Symptoms without pathology: should we try a little tenderness? PMID- 12759468 TI - Human immunodeficiency virus 1-associated minor motor disorders: perfusion weighted MR imaging and H MR spectroscopy. AB - PURPOSE: To investigate whether advanced magnetic resonance (MR) imaging techniques such as diffusion-weighted (DW) and perfusion-weighted (PW) MR imaging and hydrogen 1 (1H) MR spectroscopy can depict functional and pathophysiologic mechanisms in patients who have minor motor deficits (MMDs) associated with human immunodeficiency virus 1 (HIV-1). MATERIALS AND METHODS: Thirty-two patients with results seropositive for HIV-1 and different degrees of HIV-1-related MMD underwent conventional brain MR imaging, as well as DW and PW MR imaging and 1H MR spectroscopy of the basal ganglia. PW MR imaging data were computed pixel by pixel for creation of time-to-peak, relative regional cerebral blood volume, and bolus amplitude parameter maps. In addition, quantitative regional cerebral blood flow (rCBF) maps were calculated with respect to the arterial input function by using the singular value decomposition algorithm. For 1H MR spectroscopy, a stimulated echo acquisition mode 20, or STEAM 20, sequence was used. Spectra were fit for determination of the signal intensities of the different metabolites. According to psychomotor testing results, patients were divided into three groups: group 1, 10 patients with normal motor function; group 2, eight patients with psychomotor slowing for the first time; and group 3, 14 patients who had had sustained pathologic psychomotor slowing for at least 6 months before the MR imaging examination. RESULTS: No patients had an abnormality at either conventional or DW MR imaging. PW MR imaging depicted significantly elevated rCBF in group 2 patients (P =.039, analysis of variance [ANOVA]) and significantly elevated myo-inositol-to-creatine ratio levels in group 3 patients (P =.020, ANOVA). CONCLUSION: Quantitative PW MR imaging and 1H MR spectroscopy can depict pathologic changes in patients who have HIV-1-related MMD but normal clinical examination and conventional MR imaging findings. PMID- 12759469 TI - Peripheral vessels: MR angiography with dedicated phased-array coil with large field-of-view adapter feasibility study. AB - At magnetic resonance (MR) angiography with conventional phased-array coils, visualization of the vascular tree from the infrarenal aorta to the pedal arch is not possible in most patients. For this purpose, the authors developed a dedicated adapter with a large field of view that allows coverage of a body length of approximately 1,380 mm. Among five patients with peripheral arteriosclerotic disease, four underwent both conventional angiography and MR angiography. One hundred fourteen vascular segments from the infrarenal aorta to the feet were evaluated. Agreement between findings at conventional angiography and those at MR angiography was 94.7% (108 of 114) for all segments (96.1% [25 of 26] in the abdomen or pelvis, 97.5% [39 of 40] in the thigh, and 91.7% [44 of 48] in the calf or foot). PMID- 12759470 TI - Mammography with computer-aided detection: reproducibility assessment initial experience. AB - PURPOSE: To examine the performance and reproducibility of a commercially available computer-aided detection (CAD) system with a set of mammograms obtained in 100 patients who had undergone biopsy after positive findings at mammography. MATERIALS AND METHODS: One hundred positive mammographic examinations (four views each), depicting 96 masses and 50 microcalcification clusters, were scanned and analyzed three times by the CAD system. Reproducibility of detection sensitivity and the individual CAD-generated cues in the three images were examined. Both abnormality- and region-based detection sensitivities were compared. RESULTS: Forty-eight (96.0%) of 50 microcalcification clusters were marked on all three images in the abnormality-based analysis. Of the remaining two clusters, one was marked in two images and one was marked in only one. The abnormality-based sensitivity for mass detection ranged from 66.7% (64 of 96) to 70.8% (68 of 96). The system generated identical patterns (including images with and those without cues) for all three images in 53.3% (213 of 400) of images. For true-positive cluster regions, 88.9% (80 of 90) were marked at the same location in all images. For true-positive mass regions, 69.5% (82 of 118) were marked at the same locations in all images. In false-positive detections, only 44.0% (81 of 184) of false-positive mass regions and 31.9% (38 of 119) of false-positive cluster regions were marked at the same locations on all three images. CONCLUSION: Reproducibility of marked regions generated by the CAD system is improved from that reported previously, largely as a result of the substantial reduction in the false-positive detection rates. Reproducibility of true-positive identification of masses remains an important issue that may have methodologic and clinical practice implications. PMID- 12759471 TI - Prospective brain imaging evaluation of children with sickle cell trait: initial observations. AB - PURPOSE: To determine whether sickle cell trait (hemoglobin AS) is associated with abnormalities in the brain of asymptomatic children. MATERIALS AND METHODS: Magnetic resonance (MR) imaging and MR angiography were performed prospectively in 26 siblings (eight girls, 18 boys; mean age, 10.5 years) of patients with sickle cell disease. Two neuroradiologists, blinded as to whether a child had hemoglobin AS or AA, reviewed images obtained in siblings. With MR imaging, lacunae, loss of white matter volume, encephalomalacia, or leukoencephalopathy was identified. With MR angiography, arterial stenosis, occlusion, or tortuosity was identified. Images with definite or possible abnormalities were mixed with randomly selected images and were referred to a third neuroradiologist for a completely blinded review. In cases in which all neuroradiologists concurred, a score was assigned that indicated the sibling had an abnormality. MR angiographic findings were assigned a score for tortuosity with a new quantitative scale. RESULTS: Among 26 siblings screened, 21 children had sickle cell trait. Among these 21 children, two had mild abnormalities at MR imaging (sample prevalence rate, 10% [95% CI: 1%, 29%]), and four had arterial tortuosity (sample prevalence rate, 19% [95% CI: 5%, 42%]). When children with sickle cell trait were compared with 31 control subjects without the trait, arterial tortuosity was significantly more common in children with sickle cell trait (P =.014). Among children with sickle cell trait, percentage of hemoglobin S was significantly greater in children who had tortuosity than percentage of hemoglobin S in children who had normal blood vessels at MR angiography (P <.03). CONCLUSION: Findings suggest that greater percentage of hemoglobin S is associated with mild vasculopathy. This vasculopathy may explain some of the excess risk of stroke among African Americans. PMID- 12759472 TI - Benign and malignant lesions of the stomach: evaluation of CT criteria for differentiation. AB - PURPOSE: To determine the sensitivity and specificity of computed tomographic (CT) criteria for differentiating benign from malignant stomach lesions in patients with a thickened gastric wall at CT. MATERIALS AND METHODS: A radiology department file search revealed 36 patients with a thickened gastric wall at CT who underwent double-contrast barium suspension upper gastrointestinal tract examinations within 6 weeks before or after CT. The authors reviewed the CT images without knowledge of the final radiologic, endoscopic, or pathologic findings to determine the degree of gastric wall thickening and the symmetry, distribution, and enhancement of the thickened wall. The sensitivity and specificity of these findings for detection of malignancy were calculated. RESULTS: Two of 36 patients had two gastric abnormalities each. The final diagnoses in the 38 cases were gastritis in 19, hiatal hernia in four, benign ulcer in three, benign (n = 3) or malignant (n = 8) gastric neoplasm in 11, and no gastric abnormality in one case. Mean wall thickness was 1.5 cm (range, 0.7 7.5 cm). The finding of gastric wall thickness of 1 cm or greater had a sensitivity of 100% but a specificity of only 42% for detection of malignant or potentially malignant stomach lesions. The finding of focal, eccentric, or enhancing wall thickening had a sensitivity of 93%, 71%, or 43%, respectively, and a specificity of 8%, 75%, or 88%, respectively, for detection of these lesions. Gastric wall thickening that was 1 cm or greater and was focal, eccentric, and enhancing had a specificity of 92% but a sensitivity of only 36% for detection of these lesions. CONCLUSION: Gastric wall thickness of 1 cm or greater at CT had a sensitivity of 100% but a specificity of less than 50% for detection of malignant or potentially malignant stomach lesions that necessitated further diagnostic evaluation. PMID- 12759473 TI - Small hepatocellular carcinoma in cirrhosis: randomized comparison of radio frequency thermal ablation versus percutaneous ethanol injection. AB - PURPOSE: To compare the effectiveness of radio-frequency (RF) thermal ablation with that of percutaneous ethanol injection (PEI) for the treatment of small hepatocellular carcinoma (HCC) in patients with cirrhosis. MATERIALS AND METHODS: A series of 102 patients with hepatic cirrhosis and either single HCC 5 cm in diameter or smaller or as many as three HCCs each 3 cm or smaller (overall number of lesions, 142) randomly received either RF ablation (n = 52) or PEI (n = 50) as the sole first-line anticancer treatment. Mean follow-up was 22.9 months +/- 9.4 (SD) in the RF group and 22.4 months +/- 8.6 in the PEI group. Prognostic value of treatment techniques was assessed with univariate and multivariate Cox proportional hazards regression models. RESULTS: One- and 2-year survival rates were 100% and 98% in the RF group and 96% and 88% in the PEI group, respectively (univariate relative risk [RR] = 0.20; 95% CI: 0.02, 1.69; P =.138). One- and 2 year local recurrence-free survival rates were 98% and 96% in the RF group and 83% and 62% in the PEI group, respectively (univariate RR = 0.17; 95% CI: 0.06, 0.51; P =.002). One- and 2-year event-free survival rates were 86% and 64% for the RF group and 77% and 43% for the PEI group, respectively (univariate RR = 0.48; 95% CI: 0.27, 0.85; P =.012). RF treatment was confirmed as an independent prognostic factor for local recurrence-free survival rates with multivariate analysis (adjusted RR = 0.20; 95% CI: 0.05, 0.73; P =.015). CONCLUSION: RF ablation is superior to PEI with respect to local recurrence-free survival rates. PMID- 12759474 TI - Severe acute respiratory syndrome: radiographic appearances and pattern of progression in 138 patients. AB - PURPOSE: To retrospectively evaluate the radiographic appearances and pattern of progression of severe acute respiratory syndrome (SARS). MATERIALS AND METHODS: Chest radiographs obtained at clinical presentation and during treatment in 138 patients with confirmed SARS (66 men, 72 women; mean age, 39 years; age range, 20 83 years) were assessed. Radiographic appearances of pulmonary parenchymal abnormality, distribution, and extent of involvement on initial chest radiographs were documented. Recognizable patterns of radiographic progression were determined by comparing the overall mean percentage of lung involvement for each patient on serial radiographs. RESULTS: Initial chest radiographs were abnormal in 108 of 138 (78.3%) patients and showed air-space opacity. Lower lung zone (70 of 108, 64.8%) and right lung (82 of 108, 75.9%) were more commonly involved. In most patients, peripheral lung involvement was more common (81 of 108, 75.0%). Unifocal involvement (59 of 108, 54.6%) was more common than multifocal or bilateral involvement. No cavitation, lymphadenopathy, or pleural effusion was demonstrated. Four patterns of radiographic progression were recognized: type 1 (initial radiographic deterioration to peak level followed by radiographic improvement) in 97 of 138 patients (70.3%), type 2 (fluctuating radiographic changes) in 24 patients (17.4%), type 3 (static radiographic appearance) in 10 patients (7.3%), and type 4 (progressive radiographic deterioration) in seven patients (5.1%). Initial focal air-space opacity in 44 of 59 patients (74.6%) progressed to unilateral multifocal or bilateral involvement during treatment. CONCLUSION: Predominant peripheral location; common progression pattern from unilateral focal air-space opacity to unilateral multifocal or bilateral involvement during treatment; and lack of cavitation, lymphadenopathy, and pleural effusion are the more distinctive radiographic findings of SARS. PMID- 12759475 TI - A ligand-binding pocket in the dengue virus envelope glycoprotein. AB - Dengue virus is an emerging global health threat. Its major envelope glycoprotein, E, mediates viral attachment and entry by membrane fusion. A crystal structure of the soluble ectodomain of E from dengue virus type 2 reveals a hydrophobic pocket lined by residues that influence the pH threshold for fusion. The pocket, which accepts a hydrophobic ligand, opens and closes through a conformational shift in a beta-hairpin at the interface between two domains. These features point to a structural pathway for the fusion-activating transition and suggest a strategy for finding small-molecule inhibitors of dengue and other flaviviruses. PMID- 12759476 TI - Pseudo-cryptic speciation in coccolithophores. AB - Coccolithophores are a group of calcifying unicellular algae that constitute a major fraction of oceanic primary productivity, play an important role in the global carbon cycle, and are key biostratigraphic marker fossils. Their taxonomy is primarily based on the morphology of the minute calcite plates, or coccoliths, covering the cell. These are diverse and include widespread fine scale variation, of which the biological/taxonomic significance is unknown. Do they represent phenotypic plasticity, genetic polymorphisms, or species-specific characters? Our research on five commonly occurring coccolithophores supports the hypothesis that such variation represents pseudocryptic speciation events, occurring between 0.3 and 12.9 million years ago from a molecular clock estimation. This finding suggests strong stabilizing selection acting on coccolithophorid phenotypes. Our results also provide strong support for the use of fine scale morphological characters of coccoliths in the fossil record to improve biostratigraphic resolution and paleoceanographic data retrieval. PMID- 12759477 TI - Familial hypertrophic cardiomyopathy mutations in troponin I (K183D, G203S, K206Q) enhance filament sliding. AB - A major cause of familial hypertrophic cardiomyopathy (FHC) is dominant mutations in cardiac sarcomeric genes. Linkage studies identified FHC-related mutations in the COOH terminus of cardiac troponin I (cTnI), a region with unknown function in Ca(2+) regulation of the heart. Using in vitro assays with recombinant rat troponin subunits, we tested the hypothesis that mutations K183Delta, G203S, and K206Q in cTnI affect Ca(2+) regulation. All three mutants enhanced Ca(2+) sensitivity and maximum speed (s(max)) of filament sliding of in vitro motility assays. Enhanced s(max) (pCa 5) was observed with rabbit skeletal and rat cardiac (alpha-MHC or beta-MHC) heavy meromyosin (HMM). We developed a passive exchange method for replacing endogenous cTn in permeabilized rat cardiac trabeculae. Ca(2+) sensitivity and maximum isometric force did not differ between preparations exchanged with cTn(cTnI,K206Q) or wild-type cTn. In both trabeculae and motility assays, there was no loss of inhibition at pCa 9. These results are consistent with COOH terminus of TnI modulating actomyosin kinetics during unloaded sliding, but not during isometric force generation, and implicate enhanced cross-bridge cycling in the cTnI-related pathway(s) to hypertrophy. PMID- 12759478 TI - The National Emphysema Treatment Trial--how strong is the evidence? PMID- 12759481 TI - Guidance concerning surgery for emphysema. PMID- 12759482 TI - Do filopodia enable the growth cone to find its way? AB - Ena/VASP family proteins are required for accurate axon pathfinding in developing Drosophila, Caenorhabditis elegans, and mouse nervous systems. Recent papers by Bear et al. and Svitkina et al. provide evidence that Ena/VASP proteins regulate actin filament elongation in the lamellipodia and filopodia of nonneuronal cells. Neuronal growth cones use filopodia as their "sensory organs" that steer the growth cone by reading guidance cues in the environment. Together, these studies suggest that Ena/VASP proteins regulate axon guidance by controlling the formation or rearrangement of growth cone filopodia. PMID- 12759483 TI - Impact of pollen on human health: more than allergen carriers? AB - The transfer of pollen from floral anther to recipient stigma is the critical reproductive event among higher plants--this is the botanical view of pollen. Proteins and glycoproteins from pollen can function as allergens, environmental molecules interacting with the human immune system to elicit an allergic response in susceptible individuals--this is how allergists and immunologists see pollen grains. Between 10 and 25% of the population now have symptoms of hay fever or allergic asthma and the incidence has more than doubled in the past three decades while the reason(s) for this increment are only hypothetical, but there is a multitude of them. Despite our natural focus on this impact of pollen on human health, pollen have to be considered in a larger context. First of all, to evaluate the bioavailability of allergens from pollen, we have to understand their function and their influence factors. Furthermore, pollen grains are not only releasing proteins eliciting specific immune responses, but they also liberate bioactive lipid mediators and this much more rapidly. And last but not least, recent observations indicate, that pollen do not only induce allergy and thus have a much broader impact on human health. This review is an attempt to favour this holistic view of pollen and their impact on human health. PMID- 12759484 TI - Isolation and characterization of two complete Ara h 2 isoforms cDNA. AB - BACKGROUND: Ara h 2 is a major peanut allergen recognized by IgE in more than 90% of patients. After electrophoretic separation the purified protein exists as a doublet, and sequences of one incomplete cDNA and one genomic clone for this allergen have been reported. METHODS: Ara h 2 isoforms were purified and analyzed by mass spectroscopy, and PCR amplification products of Ara h 2 were cloned and sequenced. RESULTS: Mass spectroscopy of purified Ara h 2 clearly identified a molecular doublet of 16,670 and 18,050 Daltons. Amplification of a peanut cDNA library using PCR primer pairs located at the amino- and carboxy-terminus revealed 2 bands separated by 50 base pairs, which we cloned and sequenced. Two types of complete cDNA clones were obtained, Ara h 2.01 and Ara h 2.02. Compared to Ara h 2.01 and the previously reported cDNA sequences, Ara h 2.02 is characterized by a 12 amino acid insertion starting at position 75 that contains a third repeat of the major IgE binding epitope DPYSPS. CONCLUSION: We demonstrated the molecular and genetic characteristics of two Ara h 2 isoforms, revealing that one, Ara h 2.02, might be the more potent allergen. PMID- 12759485 TI - Oral allergy syndrome to chicory associated with birch pollen allergy. AB - BACKGROUND: A few cases of IgE-mediated chicory allergy with oral, cutaneous, and/or respiratory symptoms are reported. We present 4 patients with inhalant birch pollen allergy and oral allergy syndrome to chicory. IgE-binding proteins in chicory and cross-reactivity with birch pollen were studied. METHODS: Chicory extract was prepared and immunoblotting was used to study IgE reactivity and cross-reactions with birch pollen. RESULTS: The pattern of IgE binding to chicory was variable among the patients, with protein bands recognized at 18, 21, 40, 52 and 71 kD. Bet v 1-like proteins were detected in chicory by monoclonal antibody binding. Chicory-birch pollen cross-reactivity, as studied in 2 patients from whom enough serum was available, could be demonstrated but did not involve the Bet v 1 protein family. In one of these cases, a 51-kD protein of birch pollen was found to be responsible for cross-reactivity. CONCLUSIONS: Chicory should be added to the list of foods that can cross-react with birch pollen and cause the birch pollen-associated oral allergy syndrome. PMID- 12759486 TI - Inflammatory role of two venom components of yellow jackets (Vespula vulgaris): a mast cell degranulating peptide mastoparan and phospholipase A1. AB - BACKGROUND: Venom sac extract of yellow jackets Vespula vulgaris was toxic in mice when injected intraperitoneally but not toxic when injected subcutaneously. Necropsy showed the toxicity to be an inflammatory response. METHODS: Venom peptide and protein fractions were tested to identify the inflammatory components. The active components were tested to establish whether they might function as adjuvant for venom protein-specific antibody response. RESULTS: Venom toxicity required the synergistic action of two venom components, a mast cell degranulating peptide mastoparan and phospholipase A1. Both components stimulated prostaglandin E(2) release from murine peritoneal cells and macrophages. Mastoparan showed a weak activity to enhance IgE and IgG1 responses to a yellow jacket venom protein Ves v 5 in BALB/c mice. It was not possible to assess the adjuvant activity of phospholipase A1 because of its suppression of Ves v 5 specific response. Melittin, a mast cell degranulating peptide from bee venom, was inactive as an adjuvant for Ves v 5-specific response. CONCLUSION: Yellow jacket venom contains two inflammatory components, mastoparan and phospholipase A1. Our findings suggest that mastoparan can function as a weak adjuvant for TH2 cell-associated antibody response. PMID- 12759487 TI - Linkage and association studies of STAT6 gene polymorphisms and allergic diseases. AB - BACKGROUND: Signal transducer and activator of transcription 6 (STAT6) is a key transcription factor involved in both interleukin-4 (IL-4) and IL-13-mediated biological responses, such as allergies. Recently, we reported that the polymorphism of the STAT6 gene exon 1 was associated with allergic diseases, while another group studied the G2964A variant of the STAT6 gene's association with atopic asthma. We undertook an association study between these variants of the STAT6 gene and allergic diseases, including atopic dermatitis, bronchial asthma, and food-related anaphylaxis in a Japanese population. METHODS: STAT6 gene polymorphisms were genotyped by polymerase chain reaction (PCR) fragment length polymorphism analysis, and PCR-SSCP analysis in 106 allergic and 66 control subjects. RESULTS: The 2964A variant was in significant linkage disequilibrium with the dinucleotide repeat polymorphism, the 13-GT repeat allele of STAT6 exon 1 (p < 0.0000000003). There was no association between the STAT6 2964A variant and allergic subjects in a Japanese population (p = 0.2724). The genotype of 13/15-GT repeat allele heterozygosity was significantly associated with allergic subjects (p = 0.0006), as previously reported. In one major genotype of the STAT6 exon 1 (15 GT repeat homozygosity), wild-type 2964G allele homozygosity was significantly associated with allergic subjects (p = 0.0382). CONCLUSIONS: Our findings indicate that in combination the dinucleotide repeat polymorphism of the STAT6 exon 1 gene and the 2964A variant may be useful markers for predicting allergic diseases in a Japanese population. PMID- 12759489 TI - Immunological and clinical changes in allergic asthmatics following treatment with omalizumab. AB - IgE plays a key role in allergic asthma. We investigated whether omalizumab treatment of patients with moderate to severe allergic asthma leads to changes in inflammatory mediators and clinical symptoms. This sub-study was conducted on 35 patients with a positive skin prick test (SPT) requiring daily administration of beclomethasone dipropionate (500-1,000 microg), who participated in a multicentre, randomised, double-blind, placebo-controlled study. Omalizumab or placebo was administered at 0.016 mg/kg/IgE every 4 weeks. Patients recorded peak expiratory flow, asthma symptom score and beta(2)-agonist use in daily diaries and spirometry was performed at each visit. beta(2)-Agonist use and SPT wheal reaction decreased significantly (p < 0.05). Circulating levels of IL-5, IL-6, IL 8, IL-10, IL-13 and s-ICAM were measured before and after 16 weeks of treatment. IL-13 and s-ICAM were measured before and after 16 weeks of treatment. IL-13 decreased significantly (p < 0.01). IL-5 and IL-8 decreased in the omalizumab group compared to baseline. The other circulating mediators did not demonstrate any changes. Histamine release was significantly reduced (p < 0.01). Airway resistance (p < 0.05) and the provocative concentration inducing a 20% decrease in FEV(1) (p < 0.05) were measured before, after 16 weeks, and 3 months after completion of treatment. Both parameters decreased significantly (p < 0.05). Peripheral eosinophil count decreased significantly compared to placebo (p < 0.01). These findings suggest that omalizumab has potential as a novel treatment for allergic asthma. PMID- 12759488 TI - Analysis of natural history of Japanese cedar pollinosis. AB - BACKGROUND: The marked increase in the incidence of Japanese cedar (Cryptomeria japonica; JC) pollinosis is a social problem in Japan. Elucidation of its natural history is, therefore, essential. METHODS: Cross-sectional and vertical-sectional studies were performed regarding the effects of aging on sensitization to Japanese cedar pollen (JCP) and development of JC pollinosis by measuring serum IgE titers to JCP and by oral examination of residents of the Maruyama Town, Chiba, Japan from 1995 to 2001. We also studied the incidence of its spontaneous remission and the background factors. RESULTS: In a vertical-sectional study, the serum IgE titer to JCP was strongly influenced by the amounts of pollen scattered. An increase in age by 6 years did not reduce the IgE titer to JCP in subjects in their 40s. However, in subjects aged 60 or more, annual differences in the JCP count did not affect serum IgE titer to JCP, which remained low even after a season with a high pollen count. In subjects with JC pollinosis aged over 40 showing a CAP RAST score of more than 2 to JCP in 1995, spontaneous remission of JC pollinosis was observed in 16.1% over a period of 6 years. Factors affecting spontaneous remission include sex, age at the time, serum IgE titer to JCP and age at first onset of JC pollinosis. CONCLUSIONS: The CAP RAST score was strongly associated with spontaneous remission in the multivariable model. PMID- 12759490 TI - The nonionic radiocontrast medium iopromide does not release endothelin-1 or activate cardiac mast cells during coronary angiography. AB - BACKGROUND: High osmolal ionic radiocontrast media (RCM) cause vascular release of endothelin-1 (ET-1) and activate mast cells. Iomeprol, a nonionic RCM, has recently been reported not to activate cardiac mast cells. This coronary angiography study was performed to extend those findings using another nonionic RCM, iopromide, and to further determine whether iopromide causes release of ET 1. METHODS: Pulmonary artery plasma ET-1, histamine and serum tryptase were determined before and 30 min following angiography with iopromide in 11 subjects. ET-1, histamine and tryptase were measured using immunoassays. RESULTS: The concentrations of ET-1 (1.36 +/- 0.66 pg/ml), histamine (2.63 +/- 1.15 nM), and beta (<1 microg/l) as well as total tryptase (8.25 +/- 4.63 microg/l) in the preangiography samples were within the normal range. Following angiography, the concentrations of ET-1 (0.95 +/- 0.80 pg/ml), histamine (3.08 +/- 1.06 nM), and beta (<1 microg/l) and total tryptase (7.00 +/- 5.56 microg/l) were not significantly different. None of the subjects demonstrated a postangiography increase in mediator concentration. CONCLUSIONS: This study demonstrates the lack of release of ET-1 by iopromide. The lack of cardiac mast cell activation by iopromide is consistent with the report that iomeprol also does not activate cardiac mast cells. PMID- 12759491 TI - 15-deoxy-delta(12,14)-prostaglandin J2 inhibits IFN-gamma-induced galectin-9 expression in cultured human umbilical vein endothelial cells. AB - BACKGROUND: Galectin-9 is involved in chemotaxis and adhesion of eosinophils, and is induced in vascular endothelial cells by interferon-gamma (IFN-gamma). 15 deoxy-delta(12,14)-prostaglandin J(2) (15d-PGJ(2)) is a ligand for peroxisome proliferator-activated receptor-gamma (PPAR-gamma), and known to modulate the expression of various genes. METHODS: We have studied the effect of 15d-PGJ(2) on the IFN-gamma-induced galectin-9 expression in human umbilical vein endothelial cells (HUVEC) in culture. RESULTS: 15d-PGJ(2) inhibited the IFN-gamma-induced galectin-9 expression in a PPAR-gamma-independent manner, and also inhibited the adhesion of EoL-1 cells to an HUVEC monolayer treated with IFN-gamma. 15d-PGJ(2) partially inhibited IFN-gamma-induced phosphorylation of STAT-1 in HUVEC. CONCLUSIONS: 15d-PGJ(2) may regulate inflammatory reactions through the inhibition of galectin-9 expression. PMID- 12759492 TI - Complement-mediated enhancement of HIV-1 neutralisation by anti-HLA antibodies derived from polytransfused patients. AB - We have recently shown that 'alloimmune sera' derived from polytransfused patients (PTP sera) are able to recognise and neutralise HIV in vitro. In this study we try to identify the protein(s), which are recognised by the PTP sera and elucidate mechanisms responsible for the neutralising capacity of these sera. The PTP sera allowed immunoprecipitation (IP) of HLA class II molecules on HIV infected cells. To detect a potential cross-reactivity of alloreactive antibodies (Ab) with the HIV envelope protein gp160 or its subunits gp120/gp41 and HLA proteins, ELISA and FACS analyses were performed. The lack of reactivity of the PTP sera against rsgp160 in ELISA or FACS analysis indicated that recognition of cells was independent of HIV infection. To clarify whether interaction of the PTP sera with target cells has any effect on the infection process, virus neutralisation assays were performed. Inhibition of HIV infection was observed only when virus was pre-incubated with the PTP sera. Complement enhanced neutralisation of HIV-1 significantly. This enhancement was not due to complement mediated lysis, because pre-incubation of the target cells with PTP sera did not inhibit HIV replication. Therefore, the neutralising effect of the Ab was due to blocking of the viral attachment/fusion process and not to negative signalling after infection. Since steric hindrance is possible only when HLA and gp120/gp41 are in close vicinity, isolation of rafts and IP assays were performed. These experiments revealed that gp120 and MHC class II molecules are indeed co localised. The close physical association of gp120/gp41 and HLA strongly supports a mechanism for neutralisation of HIV by anti-HLA-Ab based on steric hindrance. PMID- 12759493 TI - RFLP analysis of mtDNA from six platyrrhine genera: phylogenetic inferences. AB - This study investigates the phylogenetic relationships of 10 species of platyrrhine primates using RFLP analysis of mtDNA. Three restriction enzymes were used to determine the restriction site haplotypes for a total of 276 individuals. Phylogenetic analysis using maximum parsimony was employed to construct phylogenetic trees. We found close phylogenetic relationships between Alouatta, Lagothrix and Ateles. We also found a close relationship between Cebus and Aotus, with Saimiri clustering with the atelines. Haplotype diversity was found in four of the species studied, in Cebus albifrons, Saimiri sciureus, Lagothrix lagotricha and Ateles fusciceps. These data provide additional information concerning the phylogenetic relationships between these platyrrhine genera and species. PMID- 12759494 TI - Role of adjuvant chemoradiotherapy for abdominal malignancies. AB - The rationale for combining surgery, radiotherapy and chemotherapy is discussed and the clinical results seen with surgery and adjuvant radiochemotherapy in three major abdominal malignancies are reviewed. A systematic approach to the literature was used. In rectal cancer, postoperative radiochemotherapy is an established treatment, although there is weak scientific support for the combined approach. The same clinical gains can also be reached much more easily with preoperative radiotherapy. In gastric cancer, a recent large randomised trial showed improved survival from postoperative radiotherapy. This was not seen in a comparably large trial in pancreatic cancer. The reasons for the different results according to primary tumour site are discussed. It is argued that adequate coverage of all adjacent regional lymph node stations is necessary for an effect on survival. PMID- 12759495 TI - A new blade for the bookwalter retractor system. AB - The table-fixed Bookwalter retractor system has been widely adopted in abdominal surgery, and during the past 15 or more years, we have succeeded in using the retractor system for thousands of cases operated on at the Taipei Veterans General Hospital, Taiwan. However, for some surgical procedures, such as gastrectomy, the blades that are currently available do not adequately hold deeper organs or tissue, such as the liver. We have successfully designed and fabricated a blade for grasping deeper tissue or organs, which is reusable and cost-effective. The shape of this new blade is described. PMID- 12759496 TI - Nodular lesion in pseudocyst following hepatic resection for hepatocellular carcinoma. PMID- 12759497 TI - Mutations in exon 11 of the c-kit gene in a myogenic tumor and a neurogenic tumor as well as in gastrointestinal stromal tumors. Utility of c-kit mutation as a prognostic biomarker for gastrointestinal mesenchymal tumor. AB - BACKGROUND/AIMS: Gain-of-function mutations in exons 9, 11 and 13 of the c-kit gene in gastrointestinal stromal tumors (GISTs) have been identified, and it has been reported that the prognosis is worse for patients with mutation-positive GISTs than for those with mutation-negative GISTs. We studied c-kit mutations in gastrointestinal mesenchymal tumors. By chance, the c-kit mutation in exon 11 was found in myogenic and neurogenic tumors as well as in GISTs. Furthermore, we studied the clinical prognostic utility of these mutations. METHODS: Ten gastrointestinal mesenchymal tumors were stained with HE and immunohistochemically analyzed with alpha-smooth muscle actin, S-100 protein, CD34 and c-kit. In these tumors, as well as in 11 cases of leiomyomas, PCR amplified DNA from the juxtamembrane (JM) domain of exon 11, the extracellular domain of exon 9 and the tyrosine kinase domain 1 of exon 13 showed a high frequency of c-kit mutation and was sequenced. RESULTS: Although c-kit mutations have previously been reported only in GISTs, we found c-kit mutations in the JM domain of exon 11 in one myogenic and one neurogenic tumor as well as in two GISTs. No c-kit mutation was seen in the 11 cases of leiomyomas. In addition, all four cases with c-kit mutation in exon 11 suffered a relapse sooner than the other cases without c-kit mutations. CONCLUSION: Clinically, the prognosis was worse for the patients with mutation-positive gastrointestinal mesenchymal tumors than for those with mutation-negative tumors. We therefore conclude that the gain of-function mutation in exon 11 of the c-kit gene is an important prognostic factor for gastrointestinal mesenchymal tumors, including myogenic and neurogenic tumors as well as GISTs. PMID- 12759480 TI - Cost effectiveness of lung-volume-reduction surgery for patients with severe emphysema. AB - BACKGROUND: The National Emphysema Treatment Trial, a randomized clinical trial comparing lung-volume-reduction surgery with medical therapy for severe emphysema, included a prospective economic analysis. METHODS: After pulmonary rehabilitation, 1218 patients at 17 medical centers were randomly assigned to lung-volume-reduction surgery or continued medical treatment. Costs for the use of medical care, medications, transportation, and time spent receiving treatment were derived from Medicare claims and data from the trial. Cost effectiveness was calculated over the duration of the trial and was estimated for 10 years of follow-up with the use of modeling based on observed trends in survival, cost, and quality of life. RESULTS: Interim analyses identified a group of patients with excess mortality and little chance of improved functional status after surgery. When these patients were excluded, the cost-effectiveness ratio for lung volume-reduction surgery as compared with medical therapy was 190,000 dollars per quality-adjusted life-year gained at 3 years and 53,000 dollars per quality adjusted life-year gained at 10 years. Subgroup analyses identified patients with predominantly upper-lobe emphysema and low exercise capacity after pulmonary rehabilitation who had lower mortality and better functional status than patients who received medical therapy. The cost-effectiveness ratio in this subgroup was 98,000 dollars per quality-adjusted life-year gained at 3 years and 21,000 dollars at 10 years. Bootstrap analysis revealed substantial uncertainty for the subgroup and 10-year estimates. CONCLUSIONS: Given its cost and benefits over three years of follow-up, lung-volume-reduction surgery is costly relative to medical therapy. Although the predictions are subject to substantial uncertainty, the procedure may be cost effective if benefits can be maintained over time. PMID- 12759479 TI - A randomized trial comparing lung-volume-reduction surgery with medical therapy for severe emphysema. AB - BACKGROUND: Lung-volume-reduction surgery has been proposed as a palliative treatment for severe emphysema. Effects on mortality, the magnitude and durability of benefits, and criteria for the selection of patients have not been established. METHODS: A total of 1218 patients with severe emphysema underwent pulmonary rehabilitation and were randomly assigned to undergo lung-volume reduction surgery or to receive continued medical treatment. RESULTS: Overall mortality was 0.11 death per person-year in both treatment groups (risk ratio for death in the surgery group, 1.01; P=0.90). After 24 months, exercise capacity had improved by more than 10 W in 15 percent of the patients in the surgery group, as compared with 3 percent of patients in the medical-therapy group (P<0.001). With the exclusion of a subgroup of 140 patients at high risk for death from surgery according to an interim analysis, overall mortality in the surgery group was 0.09 death per person-year, as compared with 0.10 death per person-year in the medical therapy group (risk ratio, 0.89; P=0.31); exercise capacity after 24 months had improved by more than 10 W in 16 percent of patients in the surgery group, as compared with 3 percent of patients in the medical-therapy group (P<0.001). Among patients with predominantly upper-lobe emphysema and low exercise capacity, mortality was lower in the surgery group than in the medical-therapy group (risk ratio for death, 0.47; P=0.005). Among patients with non-upper-lobe emphysema and high exercise capacity, mortality was higher in the surgery group than in the medical-therapy group (risk ratio, 2.06; P=0.02). CONCLUSIONS: Overall, lung volume-reduction surgery increases the chance of improved exercise capacity but does not confer a survival advantage over medical therapy. It does yield a survival advantage for patients with both predominantly upper-lobe emphysema and low base-line exercise capacity. Patients previously reported to be at high risk and those with non-upper-lobe emphysema and high base-line exercise capacity are poor candidates for lung-volume-reduction surgery, because of increased mortality and negligible functional gain. PMID- 12759498 TI - Curative surgery for local pelvic recurrence of rectal cancer. AB - BACKGROUND/AIMS: Local pelvic recurrence of rectal cancer after radical resection has been associated with morbidity and cancer-related death. This study retrospectively evaluated outcome following curative resection for rectal cancer recurring after surgery on the basis of prognosis, type of procedure and perioperative morbidity. METHODS: A total of 85 consecutive patients with local pelvic recurrence of rectal cancer were evaluated. Of these, 43 underwent microscopic curative surgery for local recurrence. Among the 43 patients, 23 underwent surgery alone and 17 received preoperative radiotherapy (40 Gy) (XRT group) in addition to the surgery. Of the 43 patients, 26 were asymptomatic. RESULTS: Curative resection was higher in the recurrences that were associated with implantation, incomplete surgical margin clearance, and intrapelvic lymph node metastasis than in other types of recurrence. With regard to surgical procedure, abdominoperineal resection (APR), with or without sacral resection, was standard following previous sphincter-preserving surgery, while total pelvic exenteration (TPE), with or without sacral resection, was common following previous APR. Local excision was not considered appropriate surgery. There was a high incidence of perioperative morbidity (64%) in patients receiving TPE. Re recurrence was observed in 18 patients (50%) after curative surgery. After a follow-up of 2 years or more, the local re-recurrence rate was 28%. The overall 5 year survival rate for patients receiving curative resection was 39%, for patients in the XRT group, 51%, and for patients in the surgery-alone group, 24% (p = 0.07). The survival rate in 26 asymptomatic patients was higher than in 17 patients with symptoms, with 5-year survival rates of 62 and 23% (p < 0.05), respectively. The cumulative local control in the preoperative radiotherapy plus en bloc surgery group (XRT group) was significantly better than in the surgery alone group (p < 0.01), and survival in the XRT group tended to be better than in surgery alone. CONCLUSIONS: These results suggest that careful patient selection according to the pattern of recurrence, area of invasion and presence of symptoms is important for successful curative surgery. Aggressive surgery with adjuvant therapy may lead to an improved salvage rate. PMID- 12759499 TI - Serum concentration of type IV collagen 7S domain as a marker for increased risk of recurrence after liver resection for hepatocellular carcinoma. AB - BACKGROUND/AIMS: Serum concentration of type IV collagen 7S domain (7S collagen) is a marker of hepatic fibrosis. We investigated the usefulness of measuring the serum 7S collagen concentration as a risk factor for recurrence after liver resection for hepatocellular carcinoma (HCC). METHODS: The serum 7S collagen concentration was measured before liver resection for HCC in 219 patients. Group 1 included 102 patients with a high serum concentration of 7S collagen (>or=8.0 ng/ml), and group 2 consisted of 117 patients with a low serum concentration of 7S collagen (<8.0 ng/ml). Clinicopathologic findings and outcome after surgery were compared between groups. RESULTS: The results of liver function tests were better in group 2 than in group 1. The percentage of patients with cirrhosis or who underwent minor resection was higher in group 1 than in group 2. The percentage of patients with moderately or poorly differentiated HCC or portal invasion was higher in group 2 than in group 1. Univariate and multivariate analyses identified that a high serum concentration of 7S collagen (>or=8.0 ng/ml) was an independent risk factor for recurrence. CONCLUSION: Measuring the serum 7S collagen concentration is useful to estimate the risk of recurrence after resection of HCC. PMID- 12759500 TI - Influence of the route of reconstruction on morbidity, mortality and local recurrence after esophagectomy for cancer. AB - BACKGROUND: A choice of retrosternal or orthotopic route for reconstruction exists after three-phase esophagectomy with cervical anastomosis. Whether the route of reconstruction affects postoperative morbidity, mortality and recurrence patterns remains controversial. STUDY DESIGN: Patients with cancer of the thoracic esophagus who underwent three-phase esophagectomy between 1990 and 1999 were studied. Postoperative outcome, recurrence patterns and survival were analyzed from a prospectively collected database. RESULTS: Seventy-five patients underwent three-phase esophagectomy. There were 46 patients in the retrosternal group and 29 in the orthotopic group. The mean age of the retrosternal group was younger than the orthotopic group, 60 and 66 years, respectively (p = 0.004). The retrosternal group also had more advanced disease; 24% of patients had curative resection compared with 59% in the orthotopic group (p = 0.003). There was no significant difference in postoperative complications except that the retrosternal group had more blood loss, median 800 ml compared with 700 ml (p = 0.04). Hospital mortality was 13% in the retrosternal group and 3.4% in the orthotopic group (p = 0.24). Multivariate analysis showed that age (odds ratio 1.16, p = 0.035) and pulmonary risk (odds ratio 10, p = 0.01) were predictive of hospital mortality, but not the route of reconstruction. No patient in the retrosternal group developed recurrence in the gastric conduit compared to 4 of 28 patients (14%) in the orthotopic group (p = 0.03). Two of these patients were symptomatic with bleeding from the intragastric recurrence. Survival was worse in the retrosternal group, 5-year survival was 29.8 vs. 8.2% (p < 0.01), reflecting the more advanced disease and higher prevalence of palliative resections. CONCLUSIONS: Cardiopulmonary complications and hospital mortality were not significantly different in the two groups. Recurrent tumor infiltration of the gastric conduit occurred in 14% of patients when the orthotopic route was used. PMID- 12759501 TI - Implications of removing a normal appendix. AB - BACKGROUND: The diagnosis of acute appendicitis remains difficult and therefore 15-30% of the removed appendices appear to be normal. The aim of this study was to investigate morbidity, mortality and costs of removing a normal appendix in patients with suspected appendicitis. PATIENTS AND METHODS: Retrospective study of patients who underwent a negative appendectomy for suspected appendicitis in the period 1991-1999 with a median follow-up of 4.4 years. Patients who underwent an elective appendectomy or appendectomy for other reasons were excluded. RESULTS: In 285 patients (70% women, 30% men) a normal appendix was removed. In 192 (67%) patients a muscle-splitting incision was performed, in 6 (2%) a median laparotomy, and in 51 (18%) the normal appendix was removed by laparoscopy. In 36 patients (13%) a diagnostic laparoscopy was converted to a muscle-splitting incision. Complications occurred in 16 (6%) patients, in 5 (2%) a re-operation was needed. The mean hospital stay was 4.4 (SE 2.8) days, in case of complications 7.4 (SE 4.2) days. The mean extra hospital costs of a negative appendectomy were EUR 2712. CONCLUSION: The removal of a normal appendix has considerable complications and costs. In an attempt to prevent these costs, extra diagnostic tools should be considered. Expensive diagnostic tools such as diagnostic laparoscopy should be used selectively in order not to further increase costs. PMID- 12759502 TI - Use of photoplethysmography to determine gastrointestinal perfusion pressure: an experimental canine model. AB - OBJECTIVES: To develop an experimental model to assess the parietal perfusion pressure (PPP) of the digestive tract using photoplethysmography. MATERIALS AND METHODS: Twenty-two mongrel dogs were used. Progressive external compression was applied to the intestinal wall and the PPP was assessed with photoplethysmography. The study group was divided into two groups. In group 1 PPP was measured at the levels of the stomach, duodenum, jejunum and transverse colon. In group 2 PPP was measured after temporary occlusion of the truncal and marginal circulation of the jejunum to provide further variables. RESULTS: The PPP decreased significantly for each successive distal section. Correlation coefficients and indices for PPP and mean arterial pressures were statistically significant (p < 0.005). Truncal occlusion provoked a drop in PPP whereas marginal occlusion scarcely modified the basal results. CONCLUSIONS: Photoplethysmography, through measurements of the residual arterial wave amplitude, is a valid method of determining quantitatively the PPP of the digestive tract and could be useful in a clinical environment. PMID- 12759503 TI - Adjuvant chemotherapy after radical resection of squamous cell carcinoma in the thoracic esophagus: who benefits? A retrospective study. AB - BACKGROUND: A definitive combined modality therapy superior to surgery alone has not yet been found for esophageal cancer. This retrospective study investigated the impact of postoperative adjuvant chemotherapy in patients who underwent curative (R0) esophagectomy with radical lymphadenectomy. STUDY DESIGN: Two hundred and eleven patients with a squamous cell carcinoma in the thoracic esophagus who underwent transthoracic curative (R0) esophagectomy with radical lymphadenectomy, such as 3-field lymphadenectomy or total 2-field lymphadenectomy, between 1988 and 2000, were retrospectively reviewed. Ninety four patients received postoperative chemotherapy - 2 courses of cisplatin (CDDP) plus fluorouracil (5-FU) or vindesine (VDS) - while the other 117 patients received surgery alone. The overall survival rate was compared between the two groups after being stratified by the numbers of the metastasis- positive lymph nodes. RESULTS: Only in the subgroup of patients with 8 or more lymph nodes metastasis- positive, the surgery-with-postoperative-chemotherapy group had a significantly better survival than the surgery-alone group. No significant difference was found in survival between the two groups in any other stratified subgroup. CONCLUSIONS: Postoperative adjuvant chemotherapy following curative (R0) esophagectomy with radical lymphadenectomy such as 3-field lymphadenectomy or total 2-field lymphadenectomy provided a benefit only in patients having metastasis in a large number - 8 or more - lymph nodes. PMID- 12759504 TI - Effects of the free radical scavenger dimethyl sulphoxide on experimental normothermic ischaemia of the liver. AB - BACKGROUND/AIMS: This study assessed the effects of intermittent or continuous hepatic ischaemia and reperfusion with or without dimethyl sulphoxide (DMSO) pre treatment in a rat ischaemic model. METHODS: One hundred and eighty rats were divided into three groups undergoing hepatic ischaemia of a total duration of 60, 90 and 120 min. Each group of rats was subdivided to receive either a continuous Pringle manoeuvre or intermittent liver pedicle clamping of 30 or 15 min. Ten minutes before ischaemia induction, 10 rats from each group were pre-treated with DMSO (500 mg/kg, b.w.) intravenously. RESULTS: With continuous hepatic pedicle clamping, survival rates inversely correlated with the duration of ischaemia, with greater survival in the intermittently clamped groups (p < 0.05). DMSO pre treatment did not affect survival but resulted in a significant reduction in liver enzyme (aspartate aminotransferase, alanine aminotransferase) release on the first postoperative day following total ischaemic times of 90 min or greater (p < 0.05). After 120 min of total ischaemia, DMSO pre-treatment resulted in higher preservation of adenosine 5'-triphosphate liver content (p < 0.05). CONCLUSION: DMSO may be used to prolong tolerance to inflow occlusion and to limit the adverse effects of ischaemia and reperfusion cycles in an experimental hepatic ischaemia model. PMID- 12759505 TI - Spontaneous endometriosis of the abdominal wall. AB - BACKGROUND: The abdominal wall is the commonest site of extrapelvic endometriosis, which usually develops in association with a prior surgical scar. CASE: A 28-year-old woman with two endometriomas in the recti abdominis, which developed in a scarless abdomen. CONCLUSIONS: Abdominal wall endometrioma can develop spontaneously in the absence of prior surgical scar and should be considered in the differential diagnosis of abdominal wall masses. PMID- 12759506 TI - A tribute to E. Bruce Hendrick. January 20, 1924 - August 17, 2001. PMID- 12759507 TI - Results of endoscopic septal fenestration in the treatment of isolated ventricular hydrocephalus. AB - A surgical series detailing the results and complications of neuroendoscopy for the treatment of isolated lateral ventricular hydrocephalus (ILVH) has yet to be presented. This retrospective case review of 32 patients examines our experience at the Primary Children's Medical Center with endoscopic fenestration of the septum pellucidum (septostomy) for ILVH. The patients who underwent endoscopic septostomy between the years of 1993 and 2001 were identified from our database. Forty-three septostomies were performed, with a mean follow-up of 30.9 months. Fifty-three percent of initial septostomies remained patent. Nine patients had a least one more septostomy performed after failure of their initial septostomy. All but one was successful. Including repeat septostomies, 81% of the patients had relief of their ILVH on the last follow-up. No septostomy failures occurred later than 6 months postoperatively. A history of multiple previous shunt procedures was highly predictive of initial septostomy failure, increasing this risk 4.5 times. Complications involved significant intraventricular hemorrhage, wound breakdown, shunt infection and sterile meningitis in four cases. We conclude that endoscopic septostomy is a reasonable treatment option for ILVH, avoiding additional shunts. Outcome is negatively affected by multiple prior shunt procedures. Favorable results can be achieved with repeat septostomies in patients who have failed prior septostomy. Lasting results are expected for septostomies that remain patent after 6 months. PMID- 12759508 TI - The influence of surgical operative experience on the duration of first ventriculoperitoneal shunt function and infection. AB - The relationship of surgeon experience, measured by operative volume, to the outcomes of ventricular shunt treatment of hydrocephalus in children is not clear. This paper explores this relationship based on first ventriculoperitoneal shunts (VPS) implanted in English-speaking Canada during the period from April 1989 to March 2001. Three thousand seven hundred and ninety-four first VPS insertions, performed by 254 surgeons, were reviewed. Surgical experience was represented by the number of shunt operations performed during the study period by each surgeon prior to the date of the operation. The 6-month shunt failure risk for less experienced surgeons was 38%, compared to 31% for more experienced surgeons. This difference decreased to 4% at 60 months and 3% at 120 months (p = 0.001). The infection rate for initial shunt insertions was 7% for patients treated by more experienced surgeons and 9.4% for those treated by less experienced surgeons (p = 0.006). A relationship between surgeon experience and shunt outcome that appears to be based on the operative experience that a surgeon brings to a procedure is in keeping with clinical experience. This observation has implications for public policy, service planning and surgical mentorship during the earlier years of a surgeon's career. PMID- 12759509 TI - Brain mapping for hemispheric tumors in children. AB - Hemispheric tumors are common in children. Pathologically, they range from indolent low grade astrocytic tumors to high grade malignant neoplasms. In general, the extent of resection correlates favorably with survival. Advances in technology are permitting surgical resection to be extended to near eloquent and eloquent regions of the brain. These techniques include advanced neuronavigation, brain mapping, and intra-operative MRI scanners. Incorporation of functional information in surgical planning should lead to safer surgical procedures with improvements in patient outcome. PMID- 12759510 TI - Pineal tumors and associated lesions: the effect of ethnicity on tumor type and treatment. AB - Pineal region lesions consist of a wide variety of rare tumor types, including deep midline cysts, intrinsic pineal tumors, germ cell tumors and vascular lesions. Advances in microsurgical, endoscopic and stereotactic techniques have helped to lower morbidity and mortality in the care of patients harboring these lesions. Surgery can be the definitive treatment in cysts and benign lesions. This report summarizes the retrospective experience of the authors with 64 pineal region and associated lesions encountered in multiple institutions over the last 20 years. Histology was obtained in 53 out of 64 radiographically apparent lesions. Direct surgical biopsy of solid and vascular tumors in the pineal region enables precise histological assessment of mixed tumors. By avoiding sampling error, precise treatment can be planned. This series, along with previously published data, shows a much higher incidence of intrinsic pineal tumors, glial tumors and nongerminomatous germ cell tumors in series from North America and Europe than in those from Japan and Korea, where germinoma is much more common. We experienced an incidence of 20.4% germinoma out of 49 solid and vascular pineal tumors, while other authors have described incidences of 51.2 and 53.5%, respectively. The fact that histology is more diverse in Western populations leads to a need to have more representative sampling. Early surgical resection combined with diversion of cerebrospinal fluid is effective in the treatment of pineal lesions and seems to be superior to the alternative of treatment based on the diagnostic response to radiation and/or on tumor markers alone. PMID- 12759512 TI - Stereotactic radiosurgery for CNS nongerminomatous germ cell tumors. Report of four cases. AB - In this study, we evaluated the results in four patients with nongerminomatous germ cell tumors (NGGCT) of the pineal region. All underwent radiosurgery in conjunction with surgical resection, fractionated radiotherapy or chemotherapy. Four male patients with pineal region NGGCT were treated with radiosurgery. The mean age was 16.5 years. Three patients had histological confirmation by stereotactic biopsy or craniotomy prior to radiosurgery. One patient was diagnosed by serum and CSF tumor markers. The mean tumor volume was 10.5 cm(3). Radiosurgery was performed with mean maximum and marginal doses of 28 and 14 Gy, respectively. At last follow-up, three patients were alive and one was dead. The mean follow-up after diagnosis and after radiosurgery was 34 and 25 months, respectively. At last follow-up, two tumors had regressed, one was unchanged and one had progressed. No patient had complications after radiosurgery. Radiosurgery can play an important adjuvant role for NGGCT patients who also undergo multimodal management. In the case of prepubertal patients, radiosurgery may play an important role by reducing the radiation dose to the surrounding normal brain. PMID- 12759511 TI - Children with growth hormone deficiency and Chiari I malformation: a morphometric analysis of the posterior cranial fossa. AB - OBJECTIVE: The posterior fossa (PF) has been found to be small in various forms of Chiari malformation. Explanations involving a connection between growth hormone deficiency (GHD) and Chiari I malformation (CIM) have been proposed. However, to date, no quantitative analysis of the PF of patients with CIM and GHD has been performed. Our study was performed to determine the geometry of the PF in children with GHD and CIM. METHODS: Morphometric analysis of the PF was performed in 10 children with GHD and CIM (group 1), 20 children with GHD and no CIM (group 2) and 50 controls. RESULTS: PF volumes for group 1 ranged from 128 to 259 +/- 33 ml, and for group 2, they ranged from 115 to 186.2 +/- 25.4 ml. Lengths of the foramen magnum for groups 1 and 2 had means of 36 and 38 mm, respectively. The mean basiocciput length and tentorial angle for groups 1 and 2 were 20 and 19 mm and 89 and 87.5 degrees, respectively. CONCLUSIONS: We have determined that children with GHD with or without CIM have no significant difference in their PF volume compared to controls. However, our data demonstrate significant underdevelopment of portions of the bony PF in both patients with GHD alone and in patients with GHD and CIM. Tentorial angles were elevated in noncontrol groups. We propose that this association is not due to an increased rate of 'midline' defects seen in GHD but rather a structurally distorted PF that is not capacious enough to house the entire developing rhombencephalon. These data will hopefully aid in the further understanding of the pathophysiology of CIM. PMID- 12759513 TI - A late result of an early operation for unilateral coronal synostosis. AB - An infant underwent strip craniectomy for unilateral coronal synostosis in 1967. This report presents the cosmetic, functional and psychosocial outcome at 34 years of age. Although aggressive, reconstructive treatment of the various forms of single-suture craniosynostosis is the current standard of care, neither the natural history of these conditions nor the long-term results of treatment have been documented systematically. PMID- 12759514 TI - Intensity of long-term treatment with warfarin is influenced by seasonal variations. AB - In a prospective study, which lasted 1 year and was performed on a large cohort of 800 patients treated at an outpatient anticoagulant clinic, we investigated possible seasonal variations in the intensity of the effect of warfarin treatment. For every season--winter, spring, summer and autumn--the mean prothrombin time reported as an international normalised ratio (INR) and the percentages of INR below 2.0 and above 4.0 were calculated. Significant seasonal variations in mean INR measurements were found, with the lowest values in summer and the highest values in autumn (F = 14.2, p < 0.0001). In addition, a trend toward a higher percentage of INR below 2.0 in summer and above 4.0 in autumn was observed. No significant differences were found between older (>65 years) and younger (<65 years) patients, although there was a trend toward more pronounced variations in younger patients. Consideration of seasonal variations might result in more accurate and safe guidance of warfarin treatment. PMID- 12759515 TI - Effect of low-dose aspirin on the international normalized ratio variability in patients with mechanical heart valve prostheses. AB - An increased risk of bleeding is associated with a more intense oral anticoagulation, a greater international normalized ratio (INR) variability and the use of aspirin. We studied the INR variability of patients (n = 121) with modern heart valves who had been prospectively randomized to receive acenocoumarol at a targeted INR of 2.4-3.6 plus aspirin 100 mg/day or acenocoumarol alone at the same dosage, to evaluate whether aspirin influences variability and thus the risk of bleeding. Variability was similar in patients with no events regardless of the use of aspirin. A statistically significantly higher variability was observed in patients with bleeding events independently of the use of aspirin. Nevertheless, the concomitant use of aspirin in patients with a high variability should be monitored closely and thoroughly. PMID- 12759516 TI - Alterations of thrombogenesis, endothelial damage and oxidative stress with reperfusion during femoral artery bypass surgery for peripheral vascular disease. AB - Peripheral vascular disease (PVD) is a significant cause of cardiovascular morbidity. We hypothesised that there would be significant alterations of thrombogenesis, platelet activation and endothelial damage, which could be associated with abnormal oxidative stress during femoral artery bypass surgery for PVD, where the femoral artery is cross-clamped (causing acute ischaemia) and reperfused (following revascularisation). To test this hypothesis, we measured sequential changes in von Willebrand factor (vWF, and index of endothelial damage/dysfunction), tissue factor (TF, an index of thrombogenesis) and soluble P selectin (sP-sel, an index of platelet activation) as well as lipid hydroperoxides (LPO, an index of oxidative stress) in 28 consecutive patients undergoing elective peripheral artery bypass surgery. Mean baseline vWF and sP sel levels in PVD patients (before clamping) were significantly higher compared with age- and sex-matched controls (unpaired t test, both p < 0.05), but there were no significant differences in TF and LPO levels. There was a correlation between TF and vWF (Spearman's, r = 0.374, p = 0.05), as well as between sP-sel and vWF at the start of surgery (r = 0.467, p = 0.012). The patients undergoing peripheral artery bypass surgery had a mean femoral artery clamp time of 28 min (standard deviation 14 min; range 11-65 min). There were no significant overall changes in sP-sel, vWF, TF and LPO with femoral artery cross-clamping and reperfusion (repeated measures ANOVA, p = NS). In conclusion, we found that during ischaemia-reperfusion during peripheral arterial bypass surgery, thrombogenesis (as measured by plasma TF) and oxidative damage (as measured by LPO) within the affected leg does not increase in the immediate perioperative period. Further studies are required to assess the mechanism(s) of ischaemia reperfusion injury in PVD, and the contributory role(s) of the endothelium and platelets. PMID- 12759517 TI - Bound thrombin from crushed clots is composed of alpha-thrombin and the N terminal regions of alpha- and gamma-chains of fibrinogen. AB - We aimed at clarifying the structural characteristics of the bound thrombin that is liberated by mechanical breakdown of fibrin clots. Fibrin clots were prepared with bovine thrombin and rabbit fibrinogen, and were crushed mechanically with a glass rod. The supernatant of the crushed clots was subjected to immunoaffinity chromatography to isolate the bound thrombin. Western blotting analysis revealed that the bound thrombin could be reacted with both antithrombin and antifibrinogen under unreduced conditions. SDS-PAGE under reduced conditions revealed that there were three bands, two of which were found to be the N terminal fragments of the alpha- and gamma-chains of fibrinogen. The bound thrombin could be dissociated into three distinct fibrin fragments and bovine alpha-thrombin when denatured by 8 M urea. Thus, the bound thrombin liberated from crushed clots is a stable complex between bovine alpha-thrombin and fibrin fragments of the N-terminal regions of rabbit alpha- and gamma-chains. PMID- 12759518 TI - Effect of phenprocoumon on monitoring of lepirudin, argatroban, melagatran and unfractionated heparin with the PiCT method. AB - Prothrombinase-induced clotting time (PiCT) is a clotting-time test for heparins and direct thrombin inhibitors to reduce drawbacks of aPTT. Effects of the direct thrombin inhibitors lepirudin, argatroban, melagatran and of unfractionated heparin (UFH) were investigated in normal and oral anticoagulant plasma samples. Lepirudin showed potentiating interferences with phenprocoumon effects. Melagatran, argatroban and UFH delivered distinct linear additive effects in both plasma sample groups. PiCT ratio reduces differences between both groups with UFH, and argatroban inhibitor-receptor-binding mode plays a role in interaction patterns. PMID- 12759520 TI - Impaired flow-mediated vasodilation in vivo and reduced shear-induced platelet reactivity in vitro in response to nitric oxide in prothrombotic, stroke-prone spontaneously hypertensive rats. AB - Previous investigations using an He-Ne laser-induced thrombosis method have shown that stroke-prone spontaneously hypertensive rats (SHRSP) have an enhanced thrombotic tendency in vivo compared to normotensive, Wistar Kyoto rats (WKY). In addition, studies in the presence of acetylcholine have suggested the presence of endothelial dysfunction in SHRSP. In contrast, shear-induced platelet reactivity in vitro appeared to be depressed in SHRSP. The aim of the present study was to investigate endothelial function in SHRSP using a new physiological in vivo model, and to determine the response of platelets to nitric oxide (NO) in non anticoagulated blood using a shear-induced platelet function in vitro method (haemostatometry). Endothelial function was estimated by measuring flow-mediated vasodilation (FMV) of the femoral artery. Vessels were exposed and blood flow was arrested using a silicone-coated arterial clamp. Vasodilation was measured by computer-assisted image analysis 3 min after release of stasis. Arterial vasodilation was observed in the femoral artery of WKY, but not in SHRSP. Vasodilation was seen in both WKY and SHRSP; however, in response to the NO donor, 1-hydroxy-2-oxo-3-(3-aminopropyl)-3-isopropyl-1-triazene (NOC 5). In contrast, 100 microM NOC 5 did not affect platelet reactivity in SHRSP. The NO scavenger, 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl 3-oxide, sodium salt (carboxy-PTIO) and the NO synthase inhibitor, N(G)-nitro-L-arginine methyl ester, hydrochloride (L-NAME), did not affect shear-induced platelet reactivity. NOC 5 at 10 microM (final concentration) inhibited shear-induced platelet reactivity in WKY. These results confirm the presence of endothelial dysfunction in SHRSP and indicate that platelets are non-responsive to NO in this hypertensive model. The data suggest that defective endothelial reactions or disturbed thrombogenic mechanisms outweigh the platelet hyporeactivity and contribute to the prothrombotic status in SHRSP. PMID- 12759519 TI - New method for the extraction of DNA from white blood cells for the detection of common genetic variants associated with thrombophilia. AB - In the last few years, FV Leiden, prothrombin 20210 and thermolabile methylene tetrahydrofolate reductase (MTHFR) 677 variants have been identified as possible risk factors associated with thrombophilia, and many PCR-based methods have been described for detecting these variations. However, the genomic DNA extraction is still a rate-limiting and time-consuming step in the PCR process. In an attempt to accelerate this procedure and make it suitable for routine laboratory, we report a single preparative technique for DNA extraction from peripheral blood samples using an anion-binding resin (GeneFizz). This method enables white blood cell lysis, DNA extraction and PCR amplification directly in the thermocycling tube on the thermocycler. The use of this new DNA extraction system coupled to a multiplex PCR allows rapid genetic screening of large cohorts of patients for thrombophilic risk factors. PMID- 12759521 TI - Plugged-percutaneous liver biopsy in patients with impaired coagulation and ascites. AB - In liver cirrhosis coagulation is impaired due to decreased synthesis of vitamin K-dependent and vitamin K-independent coagulation factors. In such patients routine liver biopsy is contraindicated due to the increased risk of bleeding. Treatment with recombinant factor VIIa or fresh frozen plasma reduces the complication rate of liver biopsy, but both have disadvantages. In this observational study, we evaluated the safety and efficacy of plugged-percutaneous liver biopsy in 36 patients with ascites (n = 9), impaired coagulation (n = 22), or both (n = 5) due to severe chronic liver disease. Among patients with clotting disorders, mean prothrombin time was 16.3 s (range 11.4-20.3) and the mean platelet count was 53 x 10(9)/l (range 19-153). Plugged-percutaneous liver biopsy was in none of the cases associated with bleeding complications (95% confidence interval 0-0.097). All biopsies were adequate for histological interpretation and therefore diagnostically successful. In our experience, plugged-percutaneous liver biopsy seems a safe and reliable method in patients with chronic liver disease associated with impaired coagulation and/or ascites needing histological evaluation. PMID- 12759522 TI - Thrombosis and antiphospholipid antibodies in non-SLE patients: predictive value of antiphosphatidylserine antibodies. PMID- 12759523 TI - Hormonal therapy in postmenopausal women with breast cancer. AB - The treatment of postmenopausal women with breast cancer presents a number of challenges. Tamoxifen has had a substantial impact on mortality in women with early-stage, estrogen-receptor-positive tumors. Despite the improvement in the treatment of breast cancer, many patients will ultimately experience a recurrence. Present treatment approaches can provide effective palliation in the advanced disease setting, but, at best, there has been a modest impact on survival. Numerous options are now available to provide effective palliation for patients with advanced disease. These options include antiestrogens, pure antiestrogens, aromatase inhibitors and progestins and LHRH agonists. Recently, several studies have reviewed the efficacy of aromatase inhibitors as first-line agents in postmenopausal women. Anastrozole and letrozole have recently been approved as first-line agents in women with metastatic breast cancer. In addition to their role in metastatic breast, trials investigating the potential of aromatase inhibitors in the early breast cancer, both in the adjuvant and neoadjuvant setting are underway. PMID- 12759524 TI - Weekly low-dose docetaxel in advanced hormone-resistant prostate cancer patients previously exposed to chemotherapy. AB - OBJECTIVE: The aim of this study was to evaluate the activity and tolerability of docetaxel in patients with hormone-resistant prostate cancer previously exposed to chemotherapy. METHODS: We enrolled 27 patients with hormone-resistant prostatic cancer that had progressed during first-line chemotherapy. The primary end-point was palliative response defined as a 2-point reduction in the 6-point present pain intensity scale, and an improvement in Karnofsky performance status of one 10-point category. The treatment consisted of weekly docetaxel 25 mg/m(2) body surface area administered by means of a 1-hour intravenous infusion with corticosteroid premedication. RESULTS: The primary criterion of palliative response was met in 13 patients (48%) after eight treatment cycles; its median duration was 6 months (range 1-8). Mean global quality of life improved in 8 and 10 patients after respectively four and eight treatment cycles. After a median follow-up of 8 months, 21 patients had died: the median survival was 9+ months (range 2-18). Weekly docetaxel was very well tolerated: grade 3 neutropenia occurred in 1 patient and grade 3 anemia in 2. CONCLUSIONS: Weekly low-dose docetaxel is an effective and well-tolerated treatment for patients with hormone resistant prostate cancer previously exposed to chemotherapy. PMID- 12759525 TI - Phase II study of irinotecan and ifosfamide in patients with advanced non-small cell lung cancer. AB - OBJECTIVES: This phase II study was conducted to investigate the efficacy and safety of irinotecan (CPT-11) and ifosfamide as first-line chemotherapy for advanced non-small cell lung cancer (NSCLC). METHODS: Eligibility criteria included histologically or cytologically confirmed NSCLC (stage IIIb or IV), no prior treatment, and measurable or evaluable disease. CPT-11 (80 mg/m(2)) was administered intravenously on days 1, 8, and 15, while ifosfamide (1.5 g/m(2)) was given on days 1 through 3 every 4 weeks. RESULTS: Forty-four patients (31 men) with a median age of 65 years (range 43-75) and a median ECOG performance status of 1 (range 0-2) were enrolled. The response rate was 29.5% [95% CI: 16.7 45.2%], with 13 partial responses. The median survival was 12.5 months, the median time to progression was 5.3 months, and the 1 and 2-year survival rates were 52.3 and 11.3%, respectively. Toxicity was generally mild; WHO grade 3-4 neutropenia was recorded in 38.6% of the patients, grade 3 diarrhea in 6.8%, and grade 3-4 nausea/vomiting in 0%. CONCLUSIONS: CPT-11 combined with ifosfamide demonstrated anti-tumor activity in advanced NSCLC, with response and survival rates similar to those of cisplatin-based chemotherapy but with a more favorable toxicity profile. PMID- 12759526 TI - A Systemic Hyperthermia Oncologic Working Group trial. Ifosfamide, carboplatin, and etoposide combined with 41.8 degrees C whole-body hyperthermia for metastatic soft tissue sarcoma. AB - BACKGROUND: Based on earlier clinical and preclinical studies, we conducted a phase II trial in metastatic sarcoma patients of the combination of 41.8 degrees C (x60 min) radiant heat (Aquatherm) whole-body hyperthermia (WBH) with 'ICE' chemotherapy. The ICE regimen consists of ifosfamide (5 g/m(2)), carboplatin (300 mg/m(2)) and etoposide (100 mg/m(2)), concurrent with WBH, with etoposide also on days 2 and 3 post-WBH. METHODS: Therapy was delivered every 4 weeks for a maximum of 4 cycles. All patients received filgrastim or lenograstim. RESULTS: Of 108 patients enrolled as of September 2001, 95 are evaluable for response. Of the evaluable patients (mean ECOG performance status approximately 1; mean age 42.3; 58% male) 33 had no prior therapy for metastatic disease, and 62 were pretreated (mean: 1.5 prior regimens). The overall response rate was 28.4% (4 complete remissions and 23 partial remissions) with stable disease (SD) in 31 patients. For no prior therapy, the response rate was 36%; in pretreated patients it was 24%. The median overall survival by Kaplan-Meier estimates was 393 days (95% CI 327, 496); the median time to treatment failure was 123 days (95% CI 77, 164). The major toxicity (287 cycles) was grade 3 or 4 neutropenia and thrombocytopenia seen in 79.7 and 60.6% of treatments respectively; there were 7 episodes of infection (grade 3/4) with 2 treatment-related deaths, bot involving disease progression and ureteral obstruction. CONCLUSION: These results are consistent with continued clinical investigation of this combined modality approach. PMID- 12759527 TI - Primary lymphoma of bone--a retrospective study. Experience at the Northern Israel Oncology Center (1979-2000). AB - OBJECTIVE: This retrospective study describes our experience with the diagnosis, treatment, results and long-term follow-up of primary bone lymphoma (PBL). PATIENTS AND METHODS: Nineteen patients diagnosed with PBL were reviewed. Seven patients presented with stage I(E) disease, four with stage II(E) (regional lymphadenopathy), and eight with stage IV disease (disseminated bone involvement). Only one stage IV patient exhibited 'B' symptoms. The majority (72%) demonstrated diffuse, large cell, B-type lymphoma. All patients were treated with adriamycin-based chemotherapy and consolidation radiotherapy to the primary site (8 patients: early PBL) or the most bulky area (3 patients: stage IV PBL). RESULTS: Ten stage I(E)/II(E) patients are alive with no evidence of disease (NED) and only one died due to metastatic secondary lung cancer while with NED from his PBL. Eight stage IV patients are alive with NED. Median follow up for all living patients: 77 months. Side effects were mild and did not necessitate delay in treatment. CONCLUSIONS: Our departmental policy of treating PBL patients with an anthracycline-based regimen and involved field radiotherapy proved to be successful in achieving excellent long-term, disease-free survival. Phase III randomized, controlled, clinical trials will determine the true role of consolidation radiotherapy in PBL, when considering severe late side effects, including radiation-induced bone tumors. PMID- 12759528 TI - Biochemotherapy for metastatic melanoma with limited central nervous system involvement. AB - OBJECTIVES: Biochemotherapy outcomes were examined in stage IV melanoma patients with previously treated or active central nervous system (CNS) metastases prior to systemic therapy. PATIENTS AND METHODS: Patients who received biochemotherapy for metastatic melanoma with active or pretreated CNS metastases were compared to patients without evidence of CNS metastases in terms of response, time to progression (TTP), overall survival (OS), and treatment toxicity. RESULTS: Twenty six (16%) of 159 total patients began biochemotherapy with previously treated or active CNS metastases (group I), compared to 133 (84%) who were radiographically free of CNS involvement (group II). A partial or complete response to biochemotherapy was seen in 13 (50%) group I patients, compared to 56 (42%) group II patients (p = 0.243). The median TTP and median survival were 5.5 and 7.0 months, respectively, for group I patients and 6.0 and 9.9 months, respectively, for group II patients (p = 0.222 and 0.434 for TTP and OS, respectively). Five (19%) group I patients survived longer than 24 months. Gamma Knife radiosurgery or surgical resection of CNS disease prior to biochemotherapy improved survival versus delayed treatment (p = 0.017 and 0.005, respectively). CONCLUSION: Patients with limited CNS metastases and widespread systemic disease can achieve prolonged survival with targeted treatment of CNS lesions and aggressive systemic therapy. PMID- 12759529 TI - 18F-fluorodeoxyglucose positron emission tomography (18F-FDG-PET) for staging and follow-up of marginal zone B-cell lymphoma. AB - OBJECTIVE: According to recent reports, nodal marginal zone lymphoma (MZL) appears to be a distinctive lymphoma entity rather than a more advanced stage of extranodal MZL of mucosa-associated lymphoid tissue (MALT). We have therefore retrospectively evaluated all patients diagnosed with nodal or extranodal MZL who have been referred to our unit for imaging using (18)F-fluorodeoxyglucose positron emission tomography ((18)F-FDG-PET). PATIENTS AND METHODS: A total of 21 patients with a diagnosis of MZL upon referral for imaging with (18)F-FDG-PET were identified. Histological reassessment of biopsy specimens confirmed the diagnosis of extranodal MZL of MALT in 14 patients, while a diagnosis of nodal MZL was verified in 6 patients. Lymphoma cell proliferation was assessed immunohistochemically using a Ki-67 antibody. Whole-body (18)F-FDG-PET scans were performed on a GE advanced PET scanner 40 min after intravenous injection of 300 380 MBq (18)F-FDG. RESULTS: None of the patients with extranodal MZL showed focal tracer uptake within verified tumor sites. In contrast, 5 of the 6 patients with nodal MZL showed significant FDG uptake within the affected lymph nodes. These results did not simply reflect the different growth fractions of the two lymphoma entities since the proliferation indices of the two groups did not differ significantly. CONCLUSION: (18)F-FDG-PET visualizes nodal MZL in a high proportion of patients whereas FDG uptake is undetectable in extranodal MZL. Although limited by the small number of patients, this study suggests that imaging with (18)F-FDG-PET might play a potential role in the diagnostic workup of patients with nodal MZL involvement. PMID- 12759530 TI - Endometrial cancer and the IGF system: a case-control study in Greece. AB - OBJECTIVES: Insulin-like growth factor I (IGF-I) and other components of the IGF system have been implicated in the etiology of several human malignancies. We have undertaken a case-control study among Greek women to explore the relation of major components of the IGF system (IGF-I, IGF-II and IGF-binding protein 3, IGFBP-3) with endometrial cancer risk. METHODS: During a 1-year period (1999), 84 incident cases of histologically confirmed endometrial cancer were compared with 84 control women admitted to the same clinical department for small gynecological operations, mainly pelvic prolapse. Data were modeled through multiple logistic regression. RESULTS: Endometrial cancer was positively associated with IGF-II and inversely with IGF-I. Both associations were statistically significant and they appear symmetrical around the null value. IGFBP-3 is positively associated with endometrial cancer risk, but this association does not reach statistical significance. CONCLUSIONS: This study adds to the gradually developing consensus that components of the IGF system play a central role in human carcinogenesis. It is possible that IGF-II, rather than IGF-I, is closely linked to the etiology of endometrial cancer, the form of cancer most strongly associated with obesity. PMID- 12759531 TI - Radiation treatment for aggressive fibromatosis: findings from observed patterns of local failure. AB - PURPOSE: We retrospectively evaluated the outcome of patients treated with radiotherapy, with or without surgery, for aggressive fibromatosis. The patterns of local failure were analyzed to determine the optimum radiation dose and volume for irradiation. PATIENTS AND METHODS: Twenty-four patients with histologically confirmed aggressive fibromatosis were treated with radiation therapy at the Yonsei Cancer Center between 1990 and 1998. The radiation dose per patient ranged from 39.6 to 59.4 Gy (mean: 49.4 Gy). The entire operative bed, with a generous margin, was included in the radiotherapy volume. Patients were considered as locally controlled if there was no evidence of the disease during their follow-up period, and if the tumors were stable for more than 2 years. The minimum duration of the follow-up period was 26 months, with a median of 69 months. RESULTS: The actuarial 10-year recurrence-free and overall survival were 88.5 and 100%, respectively. Patients who had a recurrence were salvaged by combined surgery and re-irradiation. Recurrences developed only in patients who had a recurrent disease after surgery and were treated with an inadequate radiation volume. In 9 patients with a gross measurable disease, there were no in-field failures; these had been treated with a median of 50.4 Gy (range: 40-60 Gy) of radiation. Eight patients with a microscopic residual disease were also locally controlled with 41.4-59.4 Gy (median: 45 Gy) of radiation. No patient has developed either secondary malignancy or any serious radiation complications. CONCLUSION: Radiotherapy for aggressive fibromatosis can be an effective treatment option for maintaining a disease-free status. As fibromatosis, with either a microscopic, or a gross residual disease, can be controlled with a moderate dose of radiation, adjuvant postoperative radiotherapy following surgical excision is recommended with the least sufficient margin to preserve good function and cosmesis. The geographic relationship may require a more precise definition; in addition, regardless of the existence of neighborhood normal tissue barriers, a wide coverage of the radiation volume may be needed. PMID- 12759532 TI - Subcutaneous granulocyte-macrophage colony-stimulating factor in mucositis induced by an adjuvant 5-fluorouracil plus leucovorin regimen. A phase II study and review of the literature. AB - OBJECTIVE: To test the activity of subcutaneous granulocyte-macrophage colony stimulating factor (GM-CSF) administration in patients with colorectal cancer treated with adjuvant 5-fluorouracil (5-FU) plus leucovorin (LV) and suffering from mucositis. METHODS: Thirty-one patients treated with adjuvant 5-FU 370 mg/m(2) and LV 100 mg/m(2) for 5 days every 4 weeks and reporting grade >/=2 mucositis participated in the study. Subcutaneous GM-CSF 4 microg/kg/day from days 6 to 10 was administered without chemotherapy dose reductions in the cycle that followed the cycle during which mucositis was reported. The disappearance of mucositis or a decrease by >/=1 grade was recorded as a therapeutic success. Baseline toxicity was: grade 2 stomatitis in 12 patients and grade 3 in 9; grade 4, 3 and 2 diarrhoea in 1, 4 and 8 patients, respectively. RESULTS: Seventy-seven GM-CSF cycles were administered. A success was achieved in 20 (64.5%) patients. The efficacy was assessable in 5 (16.1%) patients with grade 2 and in 8 (25.8%) with grade 3 stomatitis, respectively, as well as in 1 (3.2%) and 5 (16.1%) patients with grade 3 and 2 diarrhoea, respectively. Success (3.2%) was reported in 2 patients suffering from both grade 2 stomatitis and diarrhoea. In 7 (22.5%) patients there was no evidence of efficacy. In 4 (12.9%) patients the treatment was stopped after the first administration of GM-CSF due to a grade 2 allergic reaction. CONCLUSIONS: The subcutaneous administration of GM-CSF relieved patients from symptoms of 5-FU/LV-induced mucositis with acceptable side effects permitting the maintenance of full-dose chemotherapy. PMID- 12759533 TI - Clinicopathological significance of Nup88 expression in patients with colorectal cancer. AB - OBJECTIVE: The nucleoporin Nup88 is overexpressed in a series of human malignancies, however, its clinicopathological significance has not been studied. Our aims were to analyze Nup88 expression in normal mucosa, primary tumors and metastases from colorectal cancer patients and further to identify relationships of Nup88 expression with clinicopathological and other factors. MATERIALS AND METHODS: Using immunohistochemistry, we investigated Nup88 expression in 198 primary colorectal tumors, 96 normal mucosa samples and 35 lymph node metastases. RESULTS: The results showed that the intensity of Nup88 expression increased from the normal mucosa to the primary tumors (p < 0.0001) and tended to increase from the primary tumors to the metastases (p = 0.15). Both primary tumors and metastases presented stronger expression in the invasive margin and vascular invaded areas. Nup88 expression was positively related to distal tumor location (p = 0.01), infiltrative growth pattern (p = 0.04) and higher proliferative activity (p = 0.04) and reversely to the grade of differentiation (p = 0.02) and apoptosis (p = 0.049). Strong expression of Nup88 predicted a worse outcome in the patients with distal tumors during the follow-up period of up to 3 years (p = 0.02). CONCLUSIONS: It seems that overexpression of Nup88 was involved in the tumorigenesis and aggressiveness of colorectal cancers, and Nup88 may be used as a prognostic factor in patients with distal tumors. PMID- 12759534 TI - Ovarian Sertoli-Leydig cell tumor, endometrioid-like yolk sac tumor, and Y chromosomal material. AB - Sertoli-Leydig cell tumors (SLCTs) are rare neoplasms, accounting for less than 0.2% of ovarian tumors. The endometrioid-like variant of yolk sac tumor (YST) is very rare, and the most extensive series reported only 8 cases. We present a case of ovarian SLCT with endometrioid-like YST in a patient with a 46,XX karyotype with Y-chromosomal material. A 26-year-old woman had undergone a right salpingo oophorectomy for SLCT with endometrioid-like YST. Chromosomal analysis revealed a 46,XX karyotype with Y-chromosomal material insertion into chromosome 1. The patient's father and sister, and 7 other paternal relatives (4 male and 3 female) presented the same chromosome variant without evidence of cancer. The YST component relapsed to the right side of the uterine wall and then metastasized to the peritoneum and liver, while SLCT was eradicated with primary surgery. Several chemotherapeutic regimens were totally ineffective to control tumor progression. She died of disease progression 54 months after the diagnosis. We adopted the policy of a close surveillance for ovarian neoplasms for the 22-year-old sister of the patient, who presented the same Y-chromosomal material in her karyotype. In very rare tumors, new methods, based on molecular and cytogenetic models, are requested to define recommended management. PMID- 12759535 TI - Heterogenous S-100B protein expression patterns in malignant melanoma and association with serum protein levels. AB - OBJECTIVE: Serum S-100B is a reliable tumor marker of malignant melanoma, but efficient use is restricted to patients with metastatic disease. Therefore, the aim of our study was to assess serum S-100B levels at different stages of malignant melanoma and to compare these levels with the expression of the S-100B phenotype in primary tumors and lymph node metastases. METHODS: Fifty-nine patients were included in this study; serum S-100B protein was measured using an immunoluminometric assay while the expression pattern in the primary tumor was determined by immunohistochemistry using an anti-S-100B monoclonal antibody. RESULTS: Serum S-100B concentrations were significantly elevated in stage III (p = 0.01) patients, with normal levels in stage I-II. The most frequent S-100B protein expression pattern of the melanoma tissue was found to be diffuse staining observed in around half of the cases (52.5%) followed by heterogeneous (30.5%) and focal patterns (17%), being independent of the stage as well as the lymph node involvement. In stage I-II patients, the various staining patterns did not correlate with the serum concentration of the S-100B protein, while in stage III patients with heterogenous or diffuse S-100B staining patterns in tumor tissue, the serum marker concentration was significantly higher (p < 0.05) than in patients with focal staining. Furthermore, S-100B staining of the melanoma tissue also differed (low/negative, medium and strong staining), and serum marker concentrations corresponded to the pattern of the staining intensity. In stage I II, only strong staining was associated with elevated serum S-100B concentrations while in stage III medium and strong staining was found to be associated with significantly higher serum marker concentrations compared to patients with tumors with low/negative staining (p < 0.05). CONCLUSIONS: In malignant melanoma characterized by focal and/or low S-100B staining in the tumor tissue determined by immunohistochemistry, S-100B monitoring in the serum may not suffice to detect disease progression. PMID- 12759536 TI - Reduced expression of GNA11 and silencing of MCT1 in human breast cancers. AB - Alteration in the methylation status of a gene is often associated with its altered expression. Based on a genome scanning technique for differences in CpG methylations, methylation-sensitive representational difference analysis, DNA fragments hypermethylated in a human breast cancer were isolated. A DNA fragment was isolated from intron 1 of guanine-nucleotide-binding protein alpha-11 (GNA11). mRNA expression of GNA11 was shown to be decreased in 10 of 16 breast cancers by RT-PCR analysis, and the immunoreactivity of the GNA11 product, Galpha11 subunit of heterotrimeric G-protein, was observed to be reduced in 14 of the 16 cancers by immunohistochemistry. Methylation of a CpG island (CGI) in the 5' region of GNA11 or that of intron 1 did not show a clear correlation with its decreased expression. Another DNA fragment was isolated from a CGI in the 5' upstream region of monocarboxylate transporter 1 (MCT1), and was methylated in 4 of 20 breast cancers. The CGI was also methylated in a human breast cancer cell line, MDA-MB-231, and quantitative RT-PCR showed that its expression was almost lost in the cell line. By treatment of the cells with a demethylating agent, 5 aza-2'-deoxycytidine, the methylation was removed and the expression was restored. GNA11 is involved in signalling of gonadotropin-releasing hormone receptor, which negatively regulates cell growth. MCT1 is involved in cellular transportation of butyrate, which induces cellular differentiation. Downregulation of these two genes was suggested to be involved in human breast cancers. PMID- 12759537 TI - Prognostic role of apoptotic, Bcl-2, c-erbB-2 and p53 tumor markers in salivary gland malignancies. AB - OBJECTIVE: The clinical characteristics and survival probability rate of 36 patients with salivary gland malignancies and 10 patients with benign salivary tumors were summarized in relation to the immunohistological analysis of the tumor, apoptotic-related markers and apoptosis rate. The expression of the markers examined - Bcl-2, c-erbB-2, p53 - was detected in paraffin sections of the tumors by the streptavidin-biotin peroxidase method following heat-induced antigen retrieval, and the apoptosis rate was determined by the TUNEL method. RESULTS: The overall 5-year survival probability was 61% for patients with malignant tumors and 100% for those with benign tumors. The survival probability of patients over 60 at diagnosis was significantly lower than that of younger patients. Patients whose malignant tumors were larger than 2 cm at diagnosis had worse survival than those with smaller tumors. The survival probability of patients whose malignant tumors were located in the submandibular glands was significantly lower than that of patients whose malignancies were located in the parotid and minor salivary glands. The survival probability of patients who demonstrated positive staining for c-erbB- 2 or TUNEL was lower than for those with negative staining. Gender, the existence of concomitant non-salivary malignancies and ethnic origin had no significant impact on survival. CONCLUSIONS: Our results demonstrated significant positive staining in the salivary tumorigenic tissue but not in the surrounding non-tumorigenic tissue examined for TUNEL, c-erbB-2, Bcl-2 and p53, pointing to a biological role for all four markers in the tumorigenic process which is yet to be elucidated. Significant reduction in survival was related to the specific location of the tumor in the submandibular gland, its size and older age of patient. Survival was also found to be significantly reduced when positive staining was demonstrated in the tumor tissue for TUNEL or c-erbB-2, more so for concomitant positive staining of both markers. Clinically, the most important result of the current study is that the survival rate of the patients examined with salivary tumors larger than 2 cm, with positive staining for both TUNEL and c-erbB-2, was 0 (p = 0.0001)! PMID- 12759538 TI - Sensitivity of non-small-cell lung cancer cell lines established from patients treated with prolonged infusions of Paclitaxel. AB - OBJECTIVE: Regimens with prolonged infusions of taxanes have been developed for patients with cancer to overcome drug resistance. Our objective of the present study was to examine the impact of prolonged exposure on the cytotoxicity of taxanes against non-small-cell lung carcinoma (NSCLC) cell lines and the clinical response and outcome of the patients. METHODS: Five cell lines (NCI-H2882, H2887, -H2973, -H3122, -H3255) were derived from previously untreated patients with NSCLC who participated in clinical trials of continuous 96-hour infusions of paclitaxel followed by bolus cisplatin. Two additional cell lines (NCI-H838, H1299) with previously published data were used as controls. Drug sensitivities were assessed by the MTS (Promega) assay. Multidrug resistance (MDR) phenotypes were assessed by quantitative real-time PCR and HER-2/NEU by both immunohistochemistry and ELISA. RESULTS: The median of mean IC(50) values of docetaxel at the exposure durations of 3, 24, 72 and 120 h were 0.52, 0.06, 0.03 and 0.06 microM, respectively. The median of mean IC(50) values of paclitaxel at the exposure duration of 3, 24, 72 and 120 h were 0.48, 0.13, 0.03 and 0.02 microM, respectively. In all cell lines studied, there was a less than 4-fold difference in the IC(50) values between docetaxel and paclitaxel at 3-, 72-, and 120-hour exposure times. The single cell line with moderate MDR1 expression (NCI H2887) was the only cell line established from a patient with progressive disease when treated with paclitaxel. CONCLUSIONS: This study demonstrates prolonged exposure to both docetaxel and paclitaxel inhibits the growth of NSCLC cell lines in similar fashion. PMID- 12759539 TI - Characteristic cytokine generation patterns in cancer cells and infiltrating lymphocytes in oral squamous cell carcinomas and the influence of chemoradiation combined with immunotherapy on these patterns. AB - OBJECTIVES: Cytokines produced by tumor cells and tumor-infiltrating lymphocytes (TIL) appear to regulate tumor cell growth and the cytotoxic activity of TIL. The objectives of the present study were to investigate cytokine generation patterns in tumor cells and TIL and to examine the influence of cancer therapy on this cytokine production and the cytotoxic activity of TIL. METHODS: We determined the levels of cytokines produced by tumor cells and TIL in vitro and measured the cytotoxic activity of TIL against Daudi cells in patients with oral squamous cell carcinoma (OSC) before and 1 week after the start of concomitant chemo-radio immunotherapy. RESULTS: Before the therapy, OSC cells generated higher levels of granulocyte-macrophage colony-stimulating factor, tumor necrosis factor-alpha (TNF-alpha) and transforming growth factor-beta (TGF-beta) than did oral keratinocytes isolated from the noninflamed gingivae of healthy individuals, but both kinds of cells generated similar levels of interleukin (IL)-1beta and IL-6. Compared with peripheral blood mononuclear cells (PBMC) of the patients, TIL produced higher levels of IL-1beta, IL-6, IL-10, TNF-alpha and TGF-beta, whereas their production of IL-12 and interferon-gamma (IFN-gamma) was only slightly higher than that in PBMC. After 1 week of therapy, the cytokine production by OSC cells had largely decreased, while the production of TNF-alpha, IFN-gamma, TGF beta and IL-12 by TIL had increased greatly, although other cytokine levels were almost constant during the investigations. The cytotoxic activity of TIL was higher than that of PBMC before the therapy, and this activity was strongly increased by 1 week of therapy. CONCLUSIONS: These results suggest that the cytokine productivities of TIL and tumor cells differ from those of PBMC and normal keratinocytes, respectively, and that chemo-radio-immunotherapy modulates in situ cytokine generation, which is advantageous for inhibition of tumor cell growth and activation of TIL. PMID- 12759540 TI - Expression of vascular endothelial growth factor D is associated with lymph node metastasis in human colorectal carcinoma. AB - OBJECTIVE: Expression of vascular endothelial growth factor (VEGF)-D by tumors is associated with metastasis to lymph nodes in mice. However, there are few reports concerning the clinical significance of VEGF-D protein in human carcinoma. METHODS: After confirming production of VEGF-D by eight colorectal carcinoma cell lines, we investigated relationships between the expression of VEGF-D protein, lymph node metastasis and postoperative survival in 83 colorectal carcinoma patients. mRNA levels in cell lines were evaluated using the real-time reverse transcriptase-polymerase chain reaction, and protein was detected by Western blotting in cell lines and by immunohistochemistry in resected tissues using an antibody recognizing the processed form of the molecule. RESULTS: Immunohistochemistry showed VEGF-D-positive staining in 26 of the 83 carcinomas (31%). There was a significant relationship between the presence of VEGF-D protein and the incidence of lymph node metastasis (p < 0.01). Multivariate logistic regression analysis revealed that VEGF-D protein expression was an independent factor affecting lymph node metastasis (p < 0.01). Nonetheless, the presence or absence of VEGF-D protein had no significant impact on the survival of the patients (p = 0.15). CONCLUSION: These results suggest that the expression of VEGF-D protein could be useful in predicting the nodal status of colorectal carcinoma patients. PMID- 12759541 TI - Reduced expression of the TSP1 gene and its association with promoter hypermethylation in gastric carcinoma. AB - Thrombospondin-1 (TSP1) is a potent peptide shown in some tumor systems to be linked with angiogenesis. Epigenetic alteration of TSP1 has been reported in various primary tumors. However, the expression pattern of TSP1 has not been characterized in gastric carcinoma. We measured levels of TSP1 mRNA expression using quantitative RT-PCR in 30 gastric carcinomas and 10 non-neoplastic mucosae. In addition, we examined the correlation of the levels of TSP1 mRNA expression levels with promoter methylation status of TSP1 monitored by methylation-specific PCR as well as P53 mutation status detected by PCR-single-strand conformation polymorphism. Promoter hypermethylation of the TSP1 gene was found in 10 (33%) of 30 gastric carcinomas, and TSP1 mRNA expression levels were associated with promoter hypermethylation of TSP1 (p = 0.017; Mann-Whitney U test). P53 mutation was found in 5 (17%) of 30 gastric carcinomas, however, TSP1 mRNA expression was not associated with P53 mutation status (p = 0.858; Mann-Whitney U test). There was no correlation between TSP1 mRNA expression levels and T grade, N grade, tumor stage, or histological type. Our results suggest that transcriptional inactivation of TSP1 by aberrant DNA methylation of the promoter region may participate partly in stomach carcinogenesis through TSP1 down-regulation. PMID- 12759542 TI - Sensitive detection of human telomerase reverse transcriptase mRNA in the serum of patients with hepatocellular carcinoma. AB - BACKGROUND/AIM: Telomerase reverse transcriptase protein (hTERT) mRNA has been reported to be detectable by reverse transcription polymerase chain reaction (RT PCR) in the serum of patients with breast cancer. We measured serum hTERT mRNA in patients with hepatocellular carcinoma (HCC), and examined its clinical usefulness. METHODS: We performed RT-PCR to detect the expression of hTERT mRNA in 78 patients with HCC, 10 with liver cirrhosis (LC), 12 with chronic hepatitis (CH), and 34 healthy individuals without any liver diseases and cancers, and statistically analyzed the association with clinical parameters which include age, sex, etiology, Child classification, underlying liver disease, biochemical data, alpha-fetoprotein (alpha-AFP) number and size of tumor, and histological differentiation of HCC regarding HCC patients. RESULTS: 70 of 78 (89.7%) patients with HCC, 7 of 10 (70.0%) with LC, and 5 of 12 (41.7%) with CH were positive for hTERT expression, whereas all healthy individuals were negative for it. A multivariate analysis showed that positivity of hTERT mRNA was independently associated with AFP, tumor size, and differentiation degree. CONCLUSIONS: These results suggest that this assay is sensitive enough to detect hTERT mRNA in serum, and that it would be applicable for early detection and diagnosis of HCC or other cancers by a quantitative method. PMID- 12759543 TI - Apoptotic index and bcl-2 expression as prognostic factors in colorectal carcinoma. AB - OBJECTIVES: To examine programmed cell death in 57 colorectal carcinomas (49 primary tumours and 8 metastases) and determine the prognostic significance of apoptosis in colorectal cancer. METHODS: Apoptotic index (AI) was ascertained by counting apoptotic bodies, using terminal deoxynucleotidyl transferase mediated digoxigenin nick end labelling (Tunel assay) and the expression of bcl-2 was examined immunohistochemically. Statistical analysis was used to test the value of clinical variables, histopathological data, AI and bcl-2 expression in predicting the clinical outcome of these patients and the survival function was calculated using the Kaplan-Meier method. RESULTS: AI was found to have a significant independent effect on survival (p = 0.0006), with lower values of AI conveying better survival. CONCLUSION: In summary, these findings reveal that AI is a useful prognostic factor in colorectal carcinoma. PMID- 12759544 TI - Expression of cancer/testis tumor associated antigens in cervical squamous cell carcinoma. AB - We investigated the expression of tumor-associated antigens (TAA) of the cancer/testis (C/T) gene family in cervical squamous cell carcinomas. First, we focused on the HeLa cervical cancer derived cell line, and we found that it expresses MAGE-A1, MAGE-A2, MAGE-A3, MAGE-A4, MAGE-A6, MAGE-A12, GAGE-3/6, LAGE 1, and PRAME genes, encoding defined C/T TAA. In contrast, no expression of MAGE A10, BAGE, GAGE-1/2, or NY-ESO-1 genes was observed. Corresponding gene products could also be detected by immunoblotting and immunocytochemistry, taking advantage of monoclonal antibodies recognizing discrete TAA. Capitalizing on these data, a monoclonal antibody predominantly recognizing MAGE-A4 TAA in paraffin-embedded sections (57B) was used to investigate the C/T gene expression in clinical tumor samples. A group of 60 patients was studied, and 57B positivity was detectable to different extents in 33% of the cases (20/60). In 13 of them (21%), staining of over 50% of the tumor cells was evident, whereas healthy cells always scored negative. Remarkably, MAGE-A4 expression was significantly (p < 0.05) more frequently detectable in poorly differentiated tumors (8/13) than in well-differentiated or moderately differentiated cancers (3/15 and 9/32, respectively) and in stage FIGO II as compared with stage FIGO Ib tumors (12/23 and 5/24, respectively, p = 0.04). Interestingly, staining was mostly nuclear in well-differentiated tumors, but involved both nuclei and cytoplasm in less differentiated cancers. Positivities of comparable frequency were also detectable in a smaller series of specimens upon staining with MAGE-A1- or NY-ESO-1/LAGE-1 specific reagents. Considering the high tumor specificity of C/T TAA, our data provide the rationale for the design of immunotherapy procedures targeting these antigens in cervical cancers. PMID- 12759545 TI - Identification of a novel homeobox-containing gene, LAGY, which is downregulated in lung cancer. AB - We have isolated a novel gene, lung cancer-associated gene Y (LAGY), by suppression subtractive hybridization. The nucleotide sequence of LAGY predicts a small protein of 73 amino acids containing a putative homeobox domain with a molecular mass of 8.1 kD. Multiple-tissue Northern blot analysis revealed that LAGY is present in human placenta, lung, brain, heart and skeletal muscle. Gene mapping locates LAGY on chromosome 4q11-13.1. The expression of LAGY mRNA was widely lost in 18 lung tumor cell lines comprising all major histological types, as shown by Northern blot analysis and semiquantitative reverse transcription polymerase chain reaction. In an investigation of 72 primary lung tumors, this gene was significantly downregulated in tumors compared to 9 normal lung tissue samples. There was a significant reduction of LAGY expression in squamous cell carcinoma (SCC; n = 27) with increasing grade and stage. No expression was detectable in two high-grade SCCs or two small cell and large cell lung carcinomas (n = 4 for each). In adenocarcinoma (n = 37), expression was reduced; however, this did not reach statistical significance. Since homeodomain containing genes are known to transcriptionally regulate key cellular processes and are associated with carcinogenesis, we suggest that LAGY might be linked to lung cancer development and progression. PMID- 12759546 TI - Brefeldin A induces apoptosis and cell cycle blockade in glioblastoma cell lines. AB - Brefeldin A (BFA), a fungal metabolite known to affect the structure and function of the Golgi apparatus, has recently been shown to induce apoptosis and cell growth inhibition in various human cell lines. Glioblastomas (GB) are cerebral tumors with poor prognosis, which display resistance to current therapies including radio- and chemotherapy. The objective of this study was to investigate BFA effects in three human GB cell lines (SA4, SA146 and U87MG cells). Compared with control cells, about 60% of cell growth inhibition was observed in BFA (100 ng/ml for 24 h)-exposed cells in the three cell lines. Furthermore, in SA4 and SA146 cells, BFA was able to induce a time- and dose-dependent apoptosis detected by DAPI staining, TUNEL assay and flow-cytometric analysis. Since p53 expression was not modified after BFA exposure, BFA-induced apoptosis may follow a p53 independent pathway, as already reported. In the same way, BFA did not alter Bcl 2, Bax and Mcl-1 expression. Cell cycle analysis revealed a cell cycle arrest in early G0/G1 phase with an increase in G0/G1 cell population (70% in control cells vs. 83% in exposed cells) associated with a decrease in the S cell population (14% in control cells vs. 5.5% in exposed cells). The Ki67 labeling index also confirmed the cell cycle blockade. Our results suggest that BFA may be a potent cell cycle modulator and inducer of apoptosis in GB cell lines, and therefore may become a promising candidate for the chemotherapeutic treatment of gliomas. PMID- 12759547 TI - Peritoneal carcinoma in a male patient. AB - Peritoneal carcinoma is a rare primary tumor, described in the literature almost exclusively in women. This report describes our clinicopathological findings in a 51-year-old male patient with peritoneal carcinoma and ascites. Pathologic studies included routine histology, immunohistochemistry and electron microscopy on biopsy and autopsy tumor tissue. After chemotherapy, the patient achieved a complete remission twice, lasting for 14 months and 8 months, respectively, and died after 3 years. His clinical course was similar to that of female patients with peritoneal carcinoma or advanced ovarian cancer. Our case confirms the existence of primary peritoneal carcinoma in males. In addition, it shows that this entity responds to the same chemotherapy as used for ovarian cancer and primary peritoneal carcinoma in females. PMID- 12759548 TI - 5-fluorouracil, folinic acid and oxaliplatin administered via hepatic arterial infusion as regional second-line therapy for advanced colorectal cancer. PMID- 12759549 TI - Phase II study of cisplatin, epirubicin, and continuous-infusion 5-fluorouracil for advanced biliary tract cancer. PMID- 12759551 TI - Is gemcitabine plus vinorelbine active in second-line chemotherapy of metastatic breast cancer? a single-center phase 2 study. PMID- 12759550 TI - Gemcitabine plus irinotecan in breast cancer patients pretreated with taxanes and anthracyclines: a multicenter phase II study. PMID- 12759552 TI - The study of cytosolic phospholipase A2 gene polymorphism in schizophrenia using eye movement disturbances as an endophenotypic marker. AB - Phospholipase A(2) (PLA2) is a key enzyme of the phospholipid metabolism which shows alteration in schizophrenia. Eye movement disturbances occur in a majority of patients with schizophrenia and in a proportion of their first-degree relatives, and they have been suggested as an endophenotypic marker in genetic studies of this illness. Here we report an association between the BAN I polymorphism of the cytosolic PLA2 gene (single nucleotide polymorphism in the first intron of the gene) and the intensity of eye movement disturbances (fixation and smooth pursuit) observed in 126 schizophrenic patients. The mean intensity of both kinds of eye movement disturbances was significantly higher in individuals homozygous for the A2 genotype compared with the remaining phenotypes. There was also a trend for greater A2 allele frequency in schizophrenic patients with a higher degree of eye movement disturbances. The relative frequency of the A2/A2 genotype was higher in patients with a greater degree of eye movement disturbances occurring during both fixation and smooth pursuit tests. Our results correspond to the other studies showing an association between the cPLA2 polymorphism and schizophrenia (predominance of the A2 allele in schizophrenic subjects). PMID- 12759553 TI - Depressed patients have higher body temperature: 5-HT transporter long promoter region effects. AB - BACKGROUND: Depression has been associated with a decrease in intracellular serotonin (5-HT) reuptake through its transporter, SERT. The 5-HT transporter long promoter region (5-HTTLPR) deletion in the SERT gene has also been associated with a decrease in 5-HT reuptake. Conversely, increases in extracellular 5-HT have been associated with increased temperature. It has not been established, however, whether body temperature in depressed patients is different from controls. Here, we hypothesized that temperature would be increased in depressed patients as well as in those with the 5-HTTLPR deletion. METHODS: A strict oral temperature protocol employed single, cross-sectional, naturalistic time-of-day temperature measures in 125 subjects (46 normal controls, 79 outpatients with major depression). Controls and depressed patients were free of psychotropic medication and classified by the Structured Clinical Interview for Psychiatric Diagnoses. Eighty-one of the subjects (68 depressed, 13 normal) were additionally genotyped for 5-HTTLPR polymorphisms. RESULTS: Depressed patients had a significantly higher uncorrected body temperature (mean +/- SD 98.38 +/- 0.61 degrees F) than controls (mean +/- SD 98.13 +/- 0.59 degrees F; F = 4.8, p = 0.03). An age (F = 14.09, p < 0.001) and time-of-day (11.4, p = 0.001) correction revealed a more robust (F = 14.02, p < 0.001) difference between depressed patients (mean +/- SD 98.44 +/- 0.55 degrees F) and controls (mean +/- SD 98.02 +/- 0.56 degrees F). When normalized for age and circadian differences between subjects, random, outpatient oral temperatures had a sensitivity of 63% and a specificity of 76% in identifying the depressed subjects from the controls. Independent of depression, subjects with the 5-HTTLPR deletion (short SERT allele) were warmer (mean +/- SD 98.33 +/- 0.65 degrees F) than those lacking the short allele on either chromosome (mean +/- SD 97.91 +/- 0.69 degrees F; F = 7.0, p = 0.01). However, the genotype did not explain the temperature differences between controls and depressed patients. CONCLUSION: This is the first demonstration of an increased daytime body temperature in cases with major depression. Subjects with a corrected temperature above 98.3 degrees F were 2.6-fold more likely to be depressed. The results may strengthen the hypothesis of an inflammatory component of depression. In addition, the findings suggest a potential link between genetic differences in 5-HT transport and body temperature. PMID- 12759554 TI - Decreased lymphocyte 3H-paroxetine binding in obsessive-compulsive disorder. AB - The pathophysiology of obsessive-compulsive disorder (OCD) is mainly focused on the serotonin (5-HT) system and transporter. The presence of this structure in blood lymphocytes prompted us to investigate it, by means of the specific binding of (3)H-paroxetine ((3)H-PAR), in a group of drug-free OCD patients as compared with healthy control subjects matched for sex and age. Lymphocyte membranes and (3)H-PAR binding were carried out according to standard protocols. The results showed that the patients had a statistically significant lower density of (3)H PAR-binding sites than the control subjects. On one hand, this finding confirms previous data of an abnormal platelet 5-HT transporter in OCD, on the other it provides the possibility to explore the regulation of this structure in this and other disorders, since lymphocytes are nucleate cells. PMID- 12759555 TI - VNTR polymorphism of tyrosine hydroxylase gene and schizophrenia in the Korean population. AB - This study aimed to test the association of the tyrosine hydroxylase (TH) gene with schizophrenia in the Korean population. 334 patients with schizophrenia and 391 healthy volunteers were included. Intron 1 VNTR polymorphism of TH gene was genotyped using polymerase chain reaction techniques. The genotype and allele distribution between patients and controls were not significantly different. However, the positive score of the Positive and Negative Syndrome Scale (PANSS) was higher in the group expressing the TH10 allele (t = 2.245, p = 0.02), although no significant differences were present in the distribution of age of onset, degree of improvement using Brief Psychiatric Rating Scale (BPRS) change, total PANSS score and negative score. This finding suggests that the TH10 allele may be a liability factor for positive schizophrenia in the Korean population. PMID- 12759556 TI - Association analysis for neuronal nitric oxide synthase gene polymorphism with major depression and fluoxetine response. AB - Nitric oxide (NO) is produced from its precursor L-arginine by the enzyme NO synthase (NOS), which includes at least three distinct isoforms - neuronal (nNOS), endothelial, and inducible NOS. Recent studies have implicated NOS in the mechanism that underlies the therapeutic efficacy of antidepressant medication. In addition, major depressive disorder (MDD) patients were found to have significantly higher plasma nitrate concentrations than normal subjects, an index of NO production, in comparison to normal subjects. In a population-based association study, we tested the hypothesis that the nNOS C276T polymorphism confers susceptibility to MDD. We also examined the association between this polymorphism and therapeutic fluoxetine response in 114 MDD patients who underwent a 4-week fluoxetine treatment. The results demonstrate that the nNOS variants are found at similar frequencies in MDD patients and healthy control subjects. Further, we did not discover any genetic variants that influenced the fluoxetine response in MDD patients treated with fluoxetine. Our findings suggest that this nNOS C276T polymorphism does not play a major role in the susceptibility to, or fluoxetine response in, MDD. However, the association between other NOS variants and MDD or antidepressant response, including sexual dysfunction, may warrant further investigation. PMID- 12759557 TI - Diffusion tensor magnetic resonance imaging of disruption of regional white matter in schizophrenia. AB - Diffusion tensor imaging provides a new approach for quantifying anisotropic diffusion of white matter in vivo. We used this technique to investigate subtle disruption of regional white matter in schizophrenia. Twelve patients with schizophrenia were compared with 11 healthy controls. Psychotic symptoms were assessed using the Positive and Negative Syndrome Scale. A significant fractional anisotropy (FA) reduction was found in all white matter regions bilaterally in schizophrenic patients. Higher FA of left frontal white matter correlated significantly with higher dosage of antipsychotic medication. These findings support the view that the pathological process is a distortion of the central nervous system myelination affecting the whole white matter. Our findings also show the effects of antipsychotics on the white matter in the left frontal region in schizophrenia. PMID- 12759558 TI - Serum cholesterol in treatment-resistant depression. AB - OBJECTIVE: Patients with major depressive disorder (MDD) may have significant differences in cholesterol levels compared with healthy controls. A previous study by our group reported that depressed patients with elevated cholesterol levels (>or=200 mg/dl) were significantly more likely to be nonresponders to fluoxetine treatment than depressed patients with nonelevated cholesterol levels. However, very little is known regarding cholesterol in patients with treatment resistant depression (TRD). The purpose of this study was to compare cholesterol levels at baseline between depressed patients with and without TRD and to test whether cholesterol levels at baseline can predict clinical response in patients with TRD treated with open-label nortriptyline (NT). METHODS: Ninety-two patients with TRD entered a 6-week open trial of NT. Baseline cholesterol levels were randomly collected for 59 of these patients. Controlling for age and gender, we compared baseline cholesterol and triglyceride levels for 35 patients with TRD who did not respond to NT with 205 non-TRD patients who responded to an 8-week open trial of fluoxetine. Furthermore, with the use of logistic regression, we tested whether baseline cholesterol levels predicted clinical response to NT in the patients with TRD. RESULTS: Patients with TRD had higher triglyceride levels at baseline compared with depressed patients without TRD. Cholesterol defined as a dichotomous variable being elevated if equal to or greater than 200 mg/dl, predicted poor response to a 6-week open trial of NT in patients with TRD. CONCLUSIONS: The results of this study confirm the relationship between hypercholesterolemia and poor outcome in the treatment of MDD for patients with TRD. PMID- 12759559 TI - Catecholamine and HPA axis dysfunction in depression: relationship with suicidal behavior. AB - A large body of evidence suggests a potential role for catecholaminergic function as a possible biological factor in the control of suicidal behavior. Recently, we have used a neuroendocrine strategy to study dopaminergic and noradrenergic activities in depressed suicide attempters. However, some problems are associated with the use of growth hormone (GH) response to catecholaminergic challenge, because GH release could be decreased by a direct effect of corticosteroids at the pituitary level. Therefore, the purpose of the present study was to assess GH response to both apomorphine, a dopaminergic agonist, and clonidine, an alpha2 adrenergic agonist, according to the dexamethasone suppression test (DST) status in a sample of 20 major depressed inpatients with a history of suicide attempt compared with nonattempters. Our results tended to show that hypercortisolemia as assessed by post-DST cortisol values did not inhibit GH response to apomorphine or clonidine, suggesting that hypothalamo-pituitary-adrenal axis overactivity does not explain the impaired GH response to apomorphine in major depressed patients with a history of suicide attempt. PMID- 12759560 TI - Study of nocturnal sleep and the carryover effects of triazolam and brotizolam using neurophysiological and subjective methods. AB - In the present study, the effects of short-acting benzodiazepines on nocturnal sleep and the carryover effects of these drugs were studied. The study involved 10 young, healthy male subjects who had given their written informed consent to participate. Either a placebo (PLA), 0.125 mg triazolam (TRZ), 0.25 mg TRZ or 0.25 mg brotizolam (BRZ) was administered to the subjects in a double-blind crossover design by randomized allocation with a single oral administration at 23.00 h. A polysomnography (PSG) was recorded for each subject from 23.00 to 07.00 h the following day. Then, the Stanford Sleepiness Scale (SSS) and Kwansei Gakuin Sleepiness Scale (KSS) were checked between 07.55 and 08.00 h, and the sleep latency test (SLT) was performed between 08.00 and 08.20 h. Event-related potentials (ERPs) were then recorded with an oddball paradigm; the reaction time (RT) was measured simultaneously. According to the PSG, treatment with 0.25 mg TRZ resulted in a statistically significant increase in the percentage of stage 2 sleep (p < 0.05) and a reduction in the percentage of rapid eye movement sleep (p < 0.05) compared with PLA. None of the drugs had any effect on the percentage of slow-wave sleep compared with PLA. With regard to carryover effects, although none of the drugs had any effect on SSS, KSS, RT or ERPs, BRZ did cause a statistically significant decrease in sleep latency (p < 0.05) compared with PLA. TRZ (0.125 and 0.25 mg) and 0.25 mg BRZ exerted different effects on SLT. We suggest that these different effects are attributable to differences in the half life of these hypnotics. PMID- 12759561 TI - Frontal brain hypoactivity as a biological substrate of anxiety in patients with panic disorders. AB - Frontal brain asymmetry is associated with differences in the basic dimensions of emotion. It seems to reflect the activation of specialized systems for avoidance withdrawal behavior. Since patients with panic disorder are characterized by having both negative emotions and avoidance-withdrawal behavior, we expected them to show greater asymmetry in the frontal hemisphere change activation. Near infrared reflection spectroscopy was recorded from the left and right frontal regions of 23 patients with panic disorder without depression and from 31 healthy control participants in the following conditions: confrontation at rest with neutral (mushroom), anxiety-relevant (spider and snake) or anxiety-irrelevant but emotionally relevant stimuli (erotic picture). Emotional states and traits were assessed by the State-Trait Anxiety Inventory. The left frontal oxyhemoglobin in patients was significantly lower than in control subjects when confronted with anxiety-relevant or anxiety-irrelevant but emotionally relevant stimuli. There was no frontal brain asymmetry when patients or control subjects observed any stimuli. These data suggest that patients with panic disorder are characterized by having a greater decrease in the activation of a left frontal avoidance withdrawal system in situations with a negative valence. The findings are interpreted as biological evidence for a disturbed cortical processing in patients with panic disorder. PMID- 12759562 TI - Glucocorticoids and inflammation revisited: the state of the art. NIH clinical staff conference. AB - Glucocorticoids have been used in the treatment of inflammatory and autoimmune diseases and to prevent graft rejection for over 50 years. These hormones exert their effects through cytoplasmic, heat shock protein-bound glucocorticoid receptors that translocate into the nucleus, where they regulate the transcriptional activity of responsive genes by binding to specific promoter DNA sequences (transactivation) or by interacting with transcription factors (transrepression). By interacting with different signaling pathways, newly characterized nuclear receptor coregulators enhance or diminish the actions of glucocorticoids, thus explaining the gene-, cell-, tissue- and context-dependent actions of glucocorticoids. Glucocorticoids modulate genes involved in the priming of the innate immune response, while their actions on the adaptive immune response are to suppress cellular [T helper (Th)1-directed] immunity and promote humoral (Th2-directed) immunity and tolerance. The past decade has produced new insights into the mechanisms of glucocorticoid sensitivity and resistance of inflammatory, autoimmune and allergic diseases. Both the quality and severity of the inflammatory stimulus, as well as the genetics and constitution of the patient, play key roles in the glucocorticoid sensitivity, dependency and resistance of these diseases. Although glucocorticoids increase susceptibility to opportunistic infections, they are also highly beneficial in the presence of serious systemic inflammation, such as that observed in septic shock and acute respiratory distress syndrome, when administered in a sustained fashion throughout the course of the disease. Glucocorticoids produce their cardiovascular, metabolic and antigrowth side effects through molecular mechanisms distinct from those involved in immunomodulation. Fortunately, the first generation of tissue- and immune- versus cardiovascular/metabolic effect selective glucocorticoids is available for study and further improvement. 'Designer' glucocorticoids promise to be a great new advance in the therapy of inflammatory diseases. PMID- 12759563 TI - Neuroimmunomodulatory effects of morphine in Leishmania donovani-infected hamsters. AB - OBJECTIVE: The effect of morphine on host defense during Leishmania donovani infection in golden hamsters was studied. METHODS: Hamsters were intracardially infected with L. donovani amastigotes and then monitored by spleen touch print microscopic examination. Morphine and naloxone were administered subcutaneously and intraperitoneally, respectively. Leukocytes were counted by a hemocytometer, and ex vivo phagocytosis was determined by the examination of stained adherent macrophages. RESULTS: Low doses of morphine, 1.75 and 2.5 mg/kg x 2, administered subcutaneously on day 0 and day 15 significantly (p < 0.05) suppressed the infection, whereas high doses (20.0 and 50.0 mg/kg x 2) exacerbated the infection. On day 30, hamsters treated with low doses of morphine showed a significant (p < 0.05) increase in the number of circulating leukocytes and the pool size and phagocytic activity of peritoneal macrophages ex vivo; in hamsters treated with high doses, all these parameters appeared to be diminished. The bone marrow of morphine-treated hamsters showed a fall in total cellularity and no change in the number of monocytes; however, in those treated with low doses, the infection was completely eliminated by day 30, and paradoxically, a significant (p < 0.05) potentiation of infection was observed in hamsters treated with high doses. The spleens of hamsters treated with both low and high doses of morphine showed a significant (p < 0.05) decrease and increase in weight, respectively; treatment with low doses also caused an almost 2-fold increase in the percentage of monocytes. Morphine apparently exerted its protective effects via naloxone sensitive opioid receptors; naloxone pretreatment did not affect the potentiation of infection. CONCLUSION: Conditional doses of morphine apparently biphasically modulated the course of L. donovani infection in hamsters, at least in part through macrophage-mediated mechanisms. PMID- 12759565 TI - Brain activation of monocyte lineage cells: brain-derived soluble factors differentially regulate BV2 microglia and peripheral macrophage immune functions. AB - Brain injury and subsequent neurodegeneration are often associated with infiltrating leukocytes and the activation of microglia as well as other infiltrating cells. However, the characteristics of activation are poorly understood. The objective of this study was to further the understanding of brain regulation of microglial activation. We used an organotypic coculture paradigm to assess how brain-derived soluble factors modulate microglia and peripheral macrophage activation through microscopy and flow cytometry techniques. In the presence of brain-derived soluble factors, the BV2 microglia cell line increased MHC II and phagocytic receptor (Fcgamma II/III) expression. The increased expression correlated with a functional increase in phagocytic activity, but did not correlate with an increase in allostimulation ability. Furthermore, this interaction was selective to an interaction between brain-derived soluble factor(s) and BV2 microglia, since it was not observed in the ANA1 macrophage cell line or in primary peritoneal macrophages. The results indicated that brain derived soluble factor(s) modulate microglial activation in a manner that is distinct from the effects on peripheral macrophages. Moreover, our results suggest that inflammatory events associated with some types of brain injury may be induced by the brain without dependence on infiltrating peripheral macrophages or T lymphocytes. PMID- 12759564 TI - Mycobacterial glycolipid cord factor trehalose 6,6'-dimycolate causes a decrease in serum cortisol during the granulomatous response. AB - Serum cortisol levels were evaluated in mice following intravenous administration of purified mycobacterial glycolipid trehalose 6,6'-dimycolate (TDM). C57BL/6 mice develop lung granulomas in response to TDM, while A/J mice are deficient in this process. Administration of TDM to C57BL/6 mice led to a rapid reduction in serum cortisol, concurrent with initiation of the granulomatous response and cytokine and chemokine mRNA induction. Cortisol levels were lowest on day 5 after TDM administration, but there was significant production of IL-6, TNF-alpha and IL-1beta messages. Granuloma formation and full immune responsiveness to TDM were only apparent upon a sufficient decrease in levels of systemic cortisol. Treatment of the C57BL/6 mice with hydrocortisone abolished inflammatory responses. Histologically nonresponding A/J mice exhibited higher constitutive serum cortisol and demonstrated different kinetics of cortisol reduction upon administration of TDM. A/J mice demonstrated hyperplastic morphology in the suprarenal gland with a high degree of vacuolization in the medullary region and activation of cells in the zona fasciculata and zona reticularis. The A/J mice were dysregulated with respect to cytokine responses thought to be necessary during granuloma formation. The high constitutive serum cortisol in the A/J mice may therefore contribute to pulmonary immunoresponsiveness and the establishment of an environment counterproductive to the initiation of granulomatous responses. The identification of a mycobacterial glycolipid able to influence serum cortisol levels is unique and is discussed in relation to immunopathology during tuberculosis disease. PMID- 12759566 TI - Brain activation of monocyte-lineage cells: involvement of interleukin-6. AB - Immune processes such as phagocytosis of debris and antigen presentation can be damaging to the function and survival of brain cells. Understanding what conditions and factors mediate immune processes in the brain is central to ameliorating the pathology associated with brain injury and disease. The elevation of secreted interleukin (IL)-6 is a common feature of brain injury and neurodegenerative pathology. Using organotypic brain slice coculture, the effects of brain-derived soluble factors on immune functions in the BV2 microglial cell line were studied. We have previously shown that brain-derived soluble factors upregulated phagocytic activity and class II major histocompatibility complex (MHC II) expression and altered BV2 morphology in a manner selective for microglia and not peripheral macrophages. The present study used IL-6 neutralizing antibody to show that brain-derived IL-6 was at least partially responsible for the brain coculture-induced upregulation of MHC II expression in the BV2 microglia. Additionally, IL-6 upregulated phagocytic activity and induced morphological changes in the BV2 cells similar to brain coculture. These effects were selective for microglia, as they were not observed in peripheral macrophage cell types. The ability of IL-6-neutralizing antibody to downregulate MHC II expression while maintaining enhanced phagocytic activity could potentially evade an antagonizing immune response associated with brain injury or disease. PMID- 12759567 TI - Anterior hypothalamic lesions inhibit antigen-induced airway eosinophilia in rats. AB - OBJECTIVE: Although previous studies have found that electrolytic lesions of the anterior hypothalamic area (AHA) resulted in the suppression of anaphylaxis, their effect on late allergic responses has scarcely been investigated. To clarify the role of the AHA on possible late asthmatic responses, including their neuroendocrinological mechanisms, we examined the effect of electrolytic AHA lesions on antigen-induced eosinophilic infiltration into the airway tract and measured the plasma corticosterone and catecholamine levels in sensitized rats, i.e. a model of bronchial asthma. METHODS: The rats were randomly divided into 3 groups, including: (1) an unoperated control group; (2) a sham AHA-lesioned group and (3) an AHA-lesioned group. Then, we investigated antigen-induced eosinophilic infiltration into right bronchoalveolar lavage fluid (BALF) and the lamina propria mucosae of the left main bronchus. RESULTS: The AHA-lesioned group showed the significantly lowest number of eosinophils in both the BALF (p < 0.01) and the main bronchus (p < 0.05). The plasma adrenaline levels in the AHA-lesioned group were significantly higher than those in the other groups (p < 0.05). No differences were found in the plasma corticosterone or noradrenaline levels among the 3 groups. CONCLUSION: This study provides evidence that AHA lesions inhibit not only anaphylaxis, but also late asthmatic response related to airway eosinophilic infiltration, possibly via an alteration of the sympathetic nervous function. PMID- 12759568 TI - Modulation of interleukin-1 receptors followed by endotoxin lipopolysaccharide treatment in the mouse AtT-20 pituitary tumor cell line. AB - OBJECTIVE: We have previously reported the characterization and regulation of interleukin-1 (IL-1) receptors utilizing [125I]IL-1 binding assay in male C57BL/6 mice and the mouse AtT-20 pituitary tumor cells. In the present study, we examine IL-1 receptors using an immunoblotting method to further characterize the mechanisms regulating the interactions of IL-1 receptors with endotoxin, lipopolysaccharide (LPS). METHODS: We established Western blotting for IL-1 receptors using AtT-20 mouse pituitary tumor cells. RESULTS: Several bands were seen; however, only the 105-kD band was neutralized with a 5-fold excess of IL-1 receptor- blocking peptides, suggesting that this band is specific for IL-1 receptors. Next, we investigated the effect of LPS and IL-1beta on IL-1 receptors. Treatment of AtT-20 cells with 0.01 microg/ml of LPS did not affect IL 1 receptors. In contrast, 1 microg/ml of LPS significantly increased IL-1 receptors in AtT-20 cells compared with the control group. In addition, [125I]IL 1beta binding was markedly increased followed by 1 microg/ml of LPS. In contrast, 1 nM recombinant human IL-1beta significantly decreased IL-1 receptors in AtT-20 cells compared with the control group although treatment of AtT-20 cells with 0.01 nM IL-1beta did not affect IL-1 receptors. LPS (0.1 and 1 microg/ml) did not affect IL-1beta concentrations in the medium of AtT-20 cell culture. IL-1beta concentrations in the homogenates from AtT-20 cells were significantly decreased after 1 microg/ml of LPS treatment but not after 0.01 microg/ml LPS. CONCLUSIONS: These data demonstrate that LPS and IL-1beta differentially modulate IL-1 receptors in AtT-20 cells and LPS-induced modulation of IL-1 receptors may provide a novel mechanism for the actions of LPS to alter pituitary function during endotoxemia. Additional in vivo studies are necessary to determine the physiological relevance of this in vitro phenomenon. PMID- 12759569 TI - Nephrotoxicity and the proximal tubule. Insights from cadmium. AB - Cadmium (Cd(2+)) is a non-essential heavy metal, which is taken up from the environment into the body through pulmonary and enteral pathways. The S1 segment of the kidney proximal tubule (PT) is a major target of chronic Cd(2+) toxicity. Renal dysfunction develops in up to 7% of the general population and in its most severe form displays major features of Fanconi syndrome, such as a defective protein, amino acid, glucose, bicarbonate and phosphate reabsorption. The major pathway for Cd(2+) uptake by PT cells (PTCs) in vivo is apical endocytosis of Cd(2+) complexed to the high-affinity metal-binding protein metallothionein (MT), which may be receptor-mediated. MT is subsequently degraded in endo-lysosomes, and Cd(2+) is liberated for translocation into the cytosolic compartment, possibly using transporters for Fe(2+), Zn(2+) or Cu(2+), such as the divalent metal transporter DMT1. Free Cd(2+) ions in the extracellular space are translocated across apical and/or basolateral PTC membranes into the cytosol via transporters, whose identity remains unknown. Cytosolic Cd(2+) generates reactive oxygen species (ROS), which deplete endogenous radical scavengers. ROS also damage a variety of transport proteins, including the Na(+)/K(+)-ATPase, which are subsequently degraded by the proteasome and endo-lysosomal proteases. Cd(2+) causes mitochondrial swelling and release of cytochrome C. If these ROS-mediated stress events are not balanced by repair processes, affected cells undergo apoptosis. But Cd(2+) also induces the upregulation of cytoprotective stress and metal-scavenging proteins, such as MT. In addition, Cd(2+) upregulates the detoxifying pump multidrug resistance P-glycoprotein, which appears to protect PTCs against Cd(2+)-induced apoptosis. Thus, Cd(2+) interferes with various cellular events ranging from mechanisms of induction of programmed cell death to activation of cell survival genes. A better understanding of the cellular mechanisms involved in Cd(2+) nephrotoxicity should provide insights into other heavy metal (e.g. Pb(2+), Hg(2+)) nephropathies and various forms of acquired Fanconi syndrome. PMID- 12759570 TI - Volume control, blood pressure and cardiovascular function. Lessons from hemodialysis treatment. AB - The balance of sodium, through its dietary intake and renal excretion, determines extracellular fluid volume (ECV). Chronic renal failure and dialysis patients present with a positive sodium balance and increased ECV. The consequences are systemic hypertension and vascular and cardiac remodelling, especially left ventricular hypertrophy (LVH). High blood pressure (BP) and LVH increase the mortality risk of dialysis patients. Correction of ECV overload with ultrafiltration has led to the dry weight concept, which is the postdialysis body weight that allows BP to remain normal until the next dialysis session, without the need for antihypertensive medication and despite interdialytic weight gain. Blood pressure is the key indicator of ECV and is used to set the body weight target at each dialysis session. Normalization of BP indicates normalization of ECV, usually with a lag-time. Shortening dialysis time, which increases intradialytic morbidity, compromises the achievement of dry weight and normalization of ECV. Such intradialytic events may also lead to an increase in dialysate sodium concentration, which causes thirst, and greater interdialytic weight gain, positive sodium balance and worsening BP. These complications have led to the development of other tools for ECV assessment, such as inferior vena cava diameter, bioimpedance, and blood volume monitoring; their usefulness is discussed. PMID- 12759572 TI - What are renal defensins defending? AB - Defense against the susceptibility and damaging effects of urinary tract infection is complex and vital, as injury can lead to progressive renal injury and chronic renal failure. Recently the defensins, a family of small cationic antimicrobial peptides found in neutrophils and renal epithelial cells, have been shown to have a number of key biological properties that equip them to undertake a pivotal role in combating infection. We describe the capability of these ubiquitous and abundant peptides in the process of innate immunity, and more recently discovered roles in the adaptive immune response to infection. Furthermore, we also discuss their potential to influence other key components of the inflammatory response to infection. Despite the current state of knowledge, we are only just beginning to understand the significance of defensins as pivotal peptides in host defence and their possibilities as therapeutic targets of the future. PMID- 12759573 TI - Targeting Ras genes in kidney disease. AB - Certain changes in cellular function are characteristic of renal disease. Foremost among these is the excessive proliferation of cells, but other phenotypic changes include dysregulated apoptosis, migration, adhesion, contraction, secretion, and receptor expression. Recent advances in cell biology have revealed an extensive role for the small monomeric GTPases of the Ras superfamily in the control of these cellular events through intracellular signalling cascades. The specific Ras genes appear to play discrete and identifiable roles in a range of complex signalling networks. These insights lead to the possibility of targeting Ras genes in a specific manner in renal therapies. For example, the process of renal cell proliferation might be sensitive to downregulation of Harvey Ras and Kirsten Ras; targeting of Rho A and related species may modulate cell migration, fibrosis, and intrarenal vasoconstriction. Possible strategies for such modulation could include the use of RNA-interacting agents such as antisense DNA and si-RNA and the use of small molecules acting on Ras directly or on related signalling molecules such as Rho kinase and Raf kinase. PMID- 12759571 TI - Low urinary albumin excretion in astronauts during space missions. AB - BACKGROUND: Physiological changes occur in man during space missions also at the renal level. Proteinuria was hypothesized for space missions but research data are missing. METHODS: Urinary albumin, as an index of proteinuria, and other variables were analyzed in 4 astronauts during space missions onboard the MIR station and on the ground (control). Mission duration before first urine collection in the four astronauts was 4, 26, 26, and 106 days, respectively. On the ground, data were collected 2 months before mission in two astronauts, 6 months after in the other astronauts. A total of twenty-two 24-hour urine collections were obtained in space (n per astronaut = 1-14) and on the ground (n per astronaut = 2-12). Urinary albumin was measured by radioimmunoassay. For each astronaut, mean of data in space and on the ground was defined as individual average. RESULTS: The individual averages of 24 h urinary albumin were lower in space than on the ground in all astronauts; the difference was significant (mean +/- SD, space and on the ground = 3.41 +/- 0.56 and 4.70 +/- 1.20 mg/24 h, p = 0.017). Dietary protein intake and 24-hour urinary urea were not significantly different between space and on the ground. CONCLUSIONS: Urinary albumin excretion is low during space mission compared to data on the ground before or after mission. Low urinary albumin excretion could be another effect of exposure to weightlessness (microgravity). PMID- 12759574 TI - Growth hormone increases low-density lipoprotein receptor and HMG-CoA reductase mRNA expression in mesangial cells. AB - A dysregulation of the negative feedback mechanism of the low-density lipoprotein receptor (LDL-r) induced by hormones and cytokines may contribute to the development of glomerular injury and specifically could underlie growth hormone (GH)-induced glomerulosclerosis. The present study investigates the role of GH in the regulation of LDL-r and HMG-CoA reductase mRNA expression in mesangial cells. Mouse mesangial cells were equilibrated in a medium containing 5% lipoprotein deprived serum (LPDS) for 48 h, prior to addition of GH (0.25 microM). Transcript levels of LDL-r, HMG-CoA reductase and GH-receptor (GH-r) were measured at days 2 and 4 and intracellular lipid content was evaluated by oil red-O staining. The addition of GH significantly increased both the LDL-r and HMG-CoA reductase transcript levels at day 2 compared to control. Oil red-O positive staining increased following the initial period of 48 h lipoprotein deprivation, but addition of GH in a subsequent 48-hour period did not alter lipid content to a measurable degree compared with control. The present study demonstrates that GH significantly increased HMG-CoA reductase and LDL-r transcript levels in mesangial cells deprived of lipoproteins suggesting that abnormal levels of GH may play a role in glomerular lipid accumulation. PMID- 12759575 TI - Mechanisms of down-regulation of the renal parathyroid hormone receptor in rats with chronic renal failure. AB - Hypocalcemia, hyperphosphatemia and resistance to the action of parathyroid hormone (PTH) are well-characterized features in advanced chronic renal failure (CRF). Their pathogenesis has been attributed to both PTH receptor (PTH-R) down regulation and postreceptor abnormalities. In this study, we examined the renal expression of the PTH-R mRNA in CRF (5/6 nephrectomy) rats. Experiments were also performed to determine whether an acidic condition and PTH itself influence PTH-R mRNA expression. RT-competitive PCR was used to examine mRNA expression, and polyclonal antibody against PTH-R was used for Western blot. PTH-R mRNA expression was abundant in glomeruli, proximal convoluted and straight tubules (PCT, PST), small in medullary and cortical thick ascending limbs, and cortical collecting ducts and not detectable in outer and inner medullary collecting ducts. The expression was significantly decreased in PCT and PST in CRF rats. Decrease in PTH-R mRNA expression was observed 1 week after the induction of CRF. PTH-R protein was decreased at 2 (-23%) and 4 (-45%) weeks in renal cortex, but not in medulla in CRF rats. PTH-R mRNA expression in PST was decreased by low pH (7.1 or 6.7) incubation compared with that at pH 7.4. PTH(1-34) (10(-9) M) increased PTH-R mRNA expression in PST from control rats by 250%. The stimulatory effect of PTH on PTH-R mRNA expression was decreased by the incubation at low pH medium. In summary, renal PTH-R is down-regulated in CRF rats. The decrease in mRNA expression in PCT and PST causes the decrease in PTH-R protein. Metabolic acidosis may participate in the down-regulation of PTH-R in early stage of CRF. This abnormality could be important in the pathogenesis of secondary hyperparathyroidism of CRF. PMID- 12759576 TI - Vitamin D receptor binding to DNA is altered without the change in its expression in human renal clear cell cancer. AB - Vitamin D co-regulates cell proliferation, differentiation and apoptosis, the processes that are disturbed in cancer tissues. It acts through the vitamin D nuclear receptor (VDR) that binds to DNA in the regulatory sequences of the target genes. As the kidney is one of the key organs for vitamin D metabolism and action, we analyzed VDR expression and its DNA binding activity in human renal clear cell cancer. 24 tumors, 24 controls that were excised from the opposite pole of the same kidney and 7 controls originating from kidneys without cancer were examined. Independently of tumor grading neither Northern blots nor immunoblotting demonstrated statistically significant differences of the mean VDR mRNA and protein amounts, respectively, in the cancer as compared to both control types. In contrast, the amount of VDR-DNA complexes was lower in 52.2% of the tumors in comparison to their corresponding controls. After normalization against VDR receptor protein amount in 34.8% of the tumors VDR-DNA binding was at least 3 4 times weaker than in the controls. However, the expression of vitamin D dependent P21 gene on the mRNA level was not decreased in these cancers. It remains to be elucidated if altered VDR function due to its impaired binding to DNA contributes to the process of tumorigenesis, and what potential vitamin D dependent mechanisms are involved in this process. PMID- 12759577 TI - Extracellular actin impairs glomerular capillary repair in experimental mesangioproliferative glomerulonephritis. AB - Exogenous administration of actin prevents tumour growth in mice by specifically antagonizing angiogenin, a potent inducer of neovascularization. To investigate whether the angiogenin/actin system is also of importance in renal disease, we examined the effect of actin during glomerular capillary repair in anti-Thy-1.1 mesangioproliferative glomerulonephritis. Male Wistar rats were injected intravenously with actin, a control protein, i.e. albumin, or vehicle alone at 8, 16, 24, 32, 40 and 48 h after disease induction. On day 8, actin-treated rats showed significantly more microaneurysms and persistent mesangiolysis as compared to both control groups. This was associated with increased proteinuria in actin treated rats. Moreover, actin-treated rats showed increased counts of glomerular macrophages (+40%) and polymorphonuclear leukocytes (+100%) on day 3 as well as a decrease in glomerular endothelial area on days 3 and 8. However, no difference in early glomerular endothelial as well as non-endothelial cell proliferation was noted in actin-treated rats as compared to controls. Actin treatment had no apparent influence on mesangial cell activation (i.e. de novo expression of alpha smooth muscle actin) or glomerular accumulation of fibronectin or type IV collagen. Additional in vitro studies demonstrated that extracellular actin inhibits the angiogenin but not VEGF(165)-induced proliferation of (glomerular) endothelial cells. Moreover, actin inhibited other, yet unidentified, serum derived angiogenic factors. In conclusion, exogenous actin impairs glomerular capillary repair in experimental mesangioproliferative glomerulonephritis possibly due to interference with angiogenic factors such as angiogenin. Our combined in vivo and in vitro observations suggest that the release of intracellular actin during mesangiolysis is an endogenous pathway by which glomerular capillary damage is augmented. PMID- 12759578 TI - Advanced IgA nephropathy: to treat or not to treat? PMID- 12759579 TI - Cystatin C: what more do we need to know? PMID- 12759580 TI - Vascular calcification in chronic renal failure. AB - The prevalence and extent of vascular calcification (VC) increases rapidly with time on dialysis. There is increasing evidence that medial calcification of conduit arteries, without intimal disease, is associated with important abnormalities of vascular compliance and increased risk of cardiovascular death. Coronary artery calcification is also common in end-stage renal disease, but further research is required to determine how much of this calcification is in the form of calcified intimal atherosclerotic plaque and how much in the tunica media. Calcific uraemic arteriolopathy causes a syndrome of ischaemic necrosis of the skin and subcutaneous tissue and appears to be increasing in incidence. At all sites, arterial calcification is a biologically controlled process, with expression in vascular smooth muscle cells of genes usually expressed in osteoblasts and the formation of hydroxyapatite. High extracellular phosphate concentration induces these phenotypic changes in vitro, and much of the clinical evidence supports hyperphosphataemia as the major driver of VC. Whether warfarin treatment plays a role, by inhibiting production of vitamin-K-dependent inhibitors of calcification in humans, remains uncertain but possible. High doses of prescribed calcium-based phosphate binders are associated with VC, whereas use of sevelamer to achieve the same serum phosphate level greatly retards progression of coronary and aortic calcification. The biological mechanism by which positive calcium balance and/or episodes of hypercalcaemia promotes VC remains unclear. Treatment of established calcific uraemic arteriolopathy consists of aggressive reduction of serum calcium x phosphate product; the roles of hyperbaric oxygen, steroid therapy, and non-warfarin anticoagulation remain uncertain. PMID- 12759581 TI - Cyclophosphamide pulse therapy in advanced progressive IgA nephropathy. AB - For advanced progressive primary IgA nephropathy (IgAN) no established therapy exists. We conducted a prospective, uncontrolled trial to evaluate the effect of intravenous cyclophosphamide pulse (CyP) therapy on the course of advanced progressive IgAN. Twenty-one patients (mean age 52 +/- 10 years; male/female 20/1) with biopsy-proven IgAN without crescentic extracapillary proliferation and a serum creatinine of more than 2.0 mg/dl and/or an increase more than 25% in the previous 3 months were included. Patients were treated with CyP (750 mg/m(2 )body surface area) every 4 weeks for 6 months and low dose oral prednisolone. The loss of renal function per year was significantly reduced from 16% before therapy to 4% after therapy (p < 0.001). A further increase >25% of serum creatinine after therapy was observed in 8 patients after 0.7 years (range 0.3-3.0 years), and 3 of these patients developed end-stage renal disease. Proteinuria decreased significantly during CyP therapy. A low nadir of white blood cell and platelet count was associated with a better renal outcome (p = 0.025). In conclusion, CyP therapy and low dose oral prednisolone is effective in preserving renal function in a subgroup of patients with advanced progressive IgAN. PMID- 12759582 TI - Cohort study of advanced IgA nephropathy: efficacy and limitations of corticosteroids with tonsillectomy. AB - BACKGROUND: Elevated serum creatinine is associated with poor outcome in IgA nephropathy (IgAN). The efficacy and limitations of corticosteroids in advanced IgAN (Cr >or=1.5 mg/dl), however, remains controversial. METHODS: We retrospectively investigated 70 patients with advanced IgAN (Cr >or=1.5 mg/dl) classified into three groups according to their treatment regimens, that is, steroid pulse with tonsillectomy, conventional steroid, and supportive therapy. We evaluated the three groups to elucidate predictors for the endpoints ESRF and doubled serum creatinine from baseline. RESULTS: Steroid pulse with tonsillectomy, conventional steroid and supportive therapy were performed in 30, 25 and 15 patients, respectively. During the mean follow-up period of 70.3 (12 137) months, 41.4% of patients reached ESRF (13.3 vs. 56.0 vs. 73.3%, p < 0.001) and 45.7% doubled serum creatinine from baseline (16.7 vs. 64.0 vs. 73.3%, p < 0.001). The incidence of ESRF in the patients treated by steroid pulse with tonsillectomy was significantly lower than the incidences in the patients treated by conventional steroid and supportive therapy at a baseline creatinine level of 1.5-2 mg/dl, but no statistical difference was observed at a level of >2 mg/dl. The Kaplan-Meier estimated probability of renal survival without ESRF was 89.2, 74.1 and 72.2% at 5 years and 82.8, 51.0 and 45.1% at 8 years, respectively (p = 0.017). The predictors for ESRF, identified in a Cox proportional hazards model, were baseline serum creatinine (p < 0.001) and interstitial infiltrate (p = 0.003). Steroid pulse with tonsillectomy also had a protective effect on the risk of reaching ESRF (p = 0.013). By target cross-stratification, the patients with baseline creatinine of 1.5-2 mg/dl who underwent steroid pulse with tonsillectomy showed a better renal survival rate than the others (p < 0.001). CONCLUSION: Steroid pulse therapy combined with tonsillectomy may be more effective than conventional steroid therapy in patients with a baseline creatinine level of 1x105 was the cause of less efficacy of therapy. PMID- 12759587 TI - [Virological and clinical aspects of multiple hepatitis virus infections: preliminary data of an italian multicentre study] AB - To evaluate the interference between HBV, HCV and HDV and the clinical impact of coinfection as compared with single HBV or HCV infection, we unrolled 618 HBsAg and/or anti-HCV positive subjects (337 with liver biopsy and 281 without liver biopsy) at their first observation at one of the seven Italian Liver Units from 1993 to 1997 (Padova, Rome, Sassari, Naples, Bari, Messina, Palermo). Serum HBV DNA by dot-blot was found more frequently in patients with HBV infection alone (52% of 133 cases) than in those with HBV-HCV coinfection (28% of 64 cases, p<0.005) or in those with HBV-HDV-HCV coinfection (12% of 25 cases, p<0.0005) or with HBV-HDV coinfection (13% of 8 cases, p<0.05). We observed a higher prevalence of HCV-RNA positive cases in the patients with HCV infection alone (91.2% of 114 cases) than in those with HBV-HCV coinfection (64.5% of 62 cases, p<0.0001) or with HBV-HDV-HCV infection (19% of 21 cases, p<0.0001). These observations suggest a reciprocal inhibition of HBV and HCV genome in multiple hepatitis viral infection. A severe liver disease was more frequently observed in patients with HBV-HCV coinfection (66%) than in those with a single HBV infection (43%, p<0.05) or HCV infection (46%, p<0.05). Anti-HCV positive/anti-HBc positive patients, lacking both HBsAg and anti-HBs, compared with the anti-HCV positive/anti HBc negative ones, more frequently showed severe clinical presentation and less frequently had a sustained response to a-IFN treatment. PMID- 12759588 TI - [Incidence of fever of difficult diagnosis observed from January 1990 to August 1998] AB - Design: We evalueted incidence, period and etiology of the fever of difficult diagnosis (FDD) among 169 cases of fever of unknown origin (FUO) observed from 1 90 to 8-98. Materials and methods: we used the general and orientative diagnostic criteria useful for identifying some pathologies and also for distinguishing between infectious and non infectious fevers. Only as last attempt to solve the problem did we adopt ex-adiuvantibus treatments. Results: FDD incidence was 29.58%; we identified four main categories after detailed diagnostic work up:1) Fictitious fevers and abitual hyperthermia; 2) Systemic vasculitis with atypical clinical features; 3) Rheumatic polymyalgia; 4) Steroid-responder fevers. Conclusion: FDD is not a rare event, but of complicated resolution, some remaining unsolved. Our experience suggests that we consider only a few elements at first to distinguish organic from sine materia fevers: age, history of recent surgery, general clinical picture and aspecific indexes of inflammation. PMID- 12759589 TI - [Extraction and characterization of the lipopolysaccharide of Bartonella quintana] AB - Bartonella quintana has been reported as the cause of trench fever, persistent endocarditis, bacteriaemia and has been isolated with an increasing incidence in clinical specimens from AIDS patients. One of the main pathogenic factors of gram negative bacteria, including B. quintana, is the lipopolysaccharide (LPS). However, very little information is available on the features of Bartonella LPS. The aim of the present study was to extract, purify and characterise B. quintana LPS. The effect of the LPS under scrutiny was also evaluated on TNFa release by means of the "in vitro" human whole blood model of sepsis. The Oklahoma strain of B. quintana was grown on sheep blood agar, at 37 C, in a moist atmosphere containing 5% carbon dioxide. Cells were harvested and washed in sterile and apyrogenic saline solution and LPS extracted following the procedure of Westphal e Jann (1965), modified by Minnick (1994). The LPS of B. quintana showed the migration pattern of a deep rough chemotype, and the chromogenic limulus amoebocyte lysate test (LAL test) revealed strong reactivity at low concentrations (6.2 pg/ml). Samples of human whole blood stimulated by 1000 ng/ml of B. quintana LPS released 1707 378 pg/ml of TNFa. PMID- 12759590 TI - [Bacterial pneumonia in HIV-infected patients] AB - This case-control study assessed risk factors and prognostic indicators of 350 episodes of bacterial pneumonia in 285 HIV-infected patients. On univariate analysis, intravenous drug abuse (p<0.001), regular cigarette smoking (p<0.001), cirrhosis (p=0.04), and history of a previous episode of pneumonia (p=0.04), were risk factors for community-acquired episodes of bacterial pneumonia, whereas length of hospitalization (p=0.01) was a risk factor only for nosocomial bacterial pneumonia. The small amount of circulating T CD4+ cells, (<100/mmc) was a risk factor in both groups of pneumonia (p<0.05). Stepwise logistic regression analysis revealed that IVDA in community-acquired episodes and low levels of circulating T CD4+ cells, both in community-acquired and hospital-acquired episodes, were independent risk factors for the development of bacterial pneumonia. The case-fatality rate observed in our study was 27%. On stepwise logistic regression analysis, T CD4+ cell counts >100/mmc (p<0.02), neutropenia (p=0.04), PO2 arterial level <70 mmHg (p=0.01), and Karnofsky score <50 (p=0.04) were independent indicators of mortality. According to a personally developed prognostic score, 211 episodes of pneumonia (60%) were classified as mild, 63 (18%) as moderate, and 76 (22%) as severe. Clinicians must carefully evaluate those variables that can influence the prognosis of bacterial pneumonia to make early identification of affected patients and to promptly establish the most appropriate therapeutic strategy in each case. PMID- 12759591 TI - [Complicated purulent meningitis as first manifestation of a hidden HIV disease] AB - A case report of severe Streptococcus pneumoniae meningitis, prompting the diagnosis of a concurrent occult HIV infection, is presented. Despite a favorable in vitro susceptibility profile of the isolated microbial strain and timely antibiotic treatment, our patient suffered from permanent neurogical sequelae (severe bilateral central hypacusia). The role of penumococcal complications in the context of HIV disease and AIDS is rewieved in light of recent finding reported in the literature. PMID- 12759592 TI - [Nosocomial sepsis due to Ochrobactrum anthropi in HIV positive patients: two case reports] AB - The first two case reports of nosocomial Ochrobactrum anthropi septicemia occurring in patients with HIV disease are presented, and discussed in light of recent evidence of non-fermenting gran-negative bacilli as emerging pathogens in hospitalized immunocompromised patients. Among patients with advanced HIV infection, O. anthropi septicemia may occur even when certain presumed risk factors (notably indwelling catheters and instrumentation) are lacking, while a low CD4+ lymphocyte count, neutropenia, and concurrent AIDS-related complications may act as predisposing conditions. Despite its low intrinsic pathogenicity, O. anthropi should be taken into consideration by both microbiologists and clinicians, due to its cumbersome identification procedures, its prevailing nosocomial occurrence, and its unpredictable antibiotic susceptibility pattern. PMID- 12759593 TI - [Bilobectomy in a brazilian infant suffering from extensive monolateral pulmonary TBC] AB - As there has recently been a significant increase in childhood tuberculosis, we considered an important contribution the clinical case of an infant from Brazil, who underwent right bilobectomy to treat a previously poorly cured enlarged pulmonary tubercular infiltrate. PMID- 12759594 TI - [Fl gge's droplets] AB - After a review of the main theories by various authors on the aetiology and transmission modes of tubercular infection, the paper deals with the discovery of the tubercular bacillus made by Koch and the experiments conducted by his co worker, Cornet, who concluded that the tubercular bacillus was more concentrated around the subject suffering from open pulmonary TBC. However, it was Flugge who eventually clarified beyond any doubt, after many experiments, the real, direct transmission mode of tuberculosis through droplets of saliva emitted by the sick and which are still known today as Flugge's droplets. PMID- 12759596 TI - Questioning current practice. PMID- 12759597 TI - Anxiety medications in palliative care. PMID- 12759598 TI - On the letters on the war. PMID- 12759599 TI - Medical malpractice crisis. PMID- 12759600 TI - No regrets. PMID- 12759601 TI - Radiation: clinical responses to radiologic incidents and emergencies. PMID- 12759602 TI - Radiologic emergency: protecting schoolchildren and the public. PMID- 12759603 TI - Improving the quality of end-of-life care. PMID- 12759604 TI - Switching from i.v. to p.o. PMID- 12759610 TI - Caring knows no gender. PMID- 12759611 TI - Iraq's people between the wars. PMID- 12759612 TI - Remember the Maine! Remember the men! PMID- 12759613 TI - Compounded drugs. Are customized prescription drugs a salvation, snake oil, or both? PMID- 12759614 TI - Are nurses ready for evidence-based practice? PMID- 12759615 TI - Chemoprevention of breast cancer: recommendations and rationale. PMID- 12759616 TI - Hospital care for elderly. PMID- 12759617 TI - Bridging the gap between nurses and the media. PMID- 12759618 TI - Environmentally preferable purchasing. PMID- 12759619 TI - Predicting outcomes--not as easy as it looks. PMID- 12759620 TI - Fetal and neonatal leukemia. AB - The biological and clinical characteristics of perinatal leukemia differ significantly from those of leukemia in older children, and the prognosis is generally bleak. Once complete remission is achieved, neonates with acute myelocytic leukemia (AML) fare much better than those with acute lymphocytic leukemia (ALL). The results of this study suggest that age, sex, type of leukemia, and cytogenetic findings have a strong influence on outcome. Neonates, particularly females, with pre-B ALL have a much worse prognosis than neonates and older children with this disease. Transient leukemia in the Down syndrome neonate is associated with significant morbidity; close follow-up is recommended for at least the first 3 years of life because of the potential of developing acute leukemia, particularly AMKL (M7). The purpose of this review is to focus on the fetus and neonate in an attempt to determine the various ways leukemia differs clinically and morphologically from the disease occurring in older infants and children and to demonstrate that certain types of leukemia have a poor prognosis compared with those occurring in older children. PMID- 12759621 TI - Genetic and epigenetic alterations of the cell cycle regulators and tumor suppressor genes in pediatric osteosarcomas. AB - PURPOSE: To analyze the genetic and epigenetic alterations affecting the RB1, TP53, p16INK4, and p21WAF1 tumor suppressor genes, loss of heterozygosity (LOH) at 3q and 18q, and the clinical variables of a series of Spanish children with osteosarcoma. These genetic changes were tested for an association with prognosis. METHODS: Peripheral blood samples and clinical data were available from 76 patients with osteosarcoma. Paired tissue was available from 41 of them. The mutation and methylation status of p16INK4, p21WAF1, TP53, and RB1 was screened as well as LOH at 3q and 18q. RESULTS: Loss of heterozygosity affecting RB1 (37.2%), TP53 (42.3%), and 18q (30.8%) and TP53 mutation (39%) were frequently encountered. TP53 mutation was associated with diagnosis at a later age. RB1 alteration was associated with reduced survival and event-free survival. The clinical variables associated with poor prognosis were the presence of metastasis at diagnosis (P = 0.035) or during treatment (P = 0.016) and the chondroblastic histologic subtype (P = 0.007); the response to induction chemotherapy (<90% necrosis) also tended to be related to poor prognosis (P = 0.08). CONCLUSIONS: RB1, TP53, and possibly other tumor suppressor genes located at 18q and other localizations are involved in pediatric osteosarcoma carcinogenesis, together with other genetic alterations not fully understood to date. Based on these results, the presence of an altered RB1 gene should be regarded as a poor prognostic factor for pediatric osteosarcoma. PMID- 12759622 TI - Audiotaping communication of the diagnosis of childhood leukemia: parents' evaluation. AB - PURPOSE: To evaluate how parents viewed the authors' practice of audiotaping the initial communication of the leukemia diagnosis and of the complete program of care, including the prognosis. METHODS: From January 1997 through December 1998, at the end of the formal communication interview, parents were asked to evaluate the audiotaping by filling out a questionnaire concerning the procedure. RESULTS: Sixty-three parents of children with leukemia participated. CONCLUSIONS: The authors' use of audiotapes was strongly supported by the parents as a supplementary intervention and has become a routine procedure. PMID- 12759623 TI - Temozolomide is active in childhood, progressive, unresectable, low-grade gliomas. AB - PURPOSE: To assess the activity and tolerability of temozolomide in children with progressive low-grade gliomas (LGGs). PATIENTS AND METHODS: The authors reviewed the records of 13 children (6 months to 19 years old) with progressive LGGs and magnetic resonance imaging evidence of unresectable tumors who were treated with temozolomide at the authors' institution since 1999. RESULTS: Four patients received a 5-day regimen of temozolomide (150 mg/m2 per day) repeated every 28 days, and nine patients received a 42-day regimen (75 mg/m2 per day) repeated every 56 days. Three patients demonstrated partial responses to temozolomide, with a median time to maximal response of 5 months (range 4-12 months), and one had a minor response at 9 months. Four patients developed progression while on temozolomide, with a median time to progression of 7 months (range 1-12 months). Five patients had disease stabilization. Among the five patients with prior chemotherapy and/or radiation therapy, temozolomide was associated with disease stabilization in three and tumor response in one. In the three patients with neurofibromatosis type 1, two patients experienced tumor responses and one disease stabilization. Thrombocytopenia, nausea, emesis, and fatigue were the most common toxicities. Four patients discontinued therapy because of the side effects. CONCLUSIONS: Temozolomide is active in children with LGGs. It is effective in previously treated patients and in patients with neurofibromatosis type 1. The 42-day regimen appears less toxic than the 5-day regimen. Any impact on survival for these patients remains to be demonstrated. PMID- 12759624 TI - Low-dose tissue plasminogen activator thrombolysis in children. AB - PURPOSE: To compare results of low-dose tissue plasminogen activator (TPA) in children with arterial and venous thrombi relative to standard published dosing. METHODS: Subjects consisted of all consecutive children with objectively confirmed thrombi for whom TPA thrombolysis was clinically ordered by the authors. Initial dosing used published standard dose (0.1-0.5 mg/kg per hour). With experience, a low-dose regimen (0.01-0.06 mg/kg per hour) was given in an attempt to derive a minimal effective dose. RESULTS: Thirty-five children were treated with TPA. Either standard or low-dose infusions of TPA resulted in complete thrombolysis of 28 of 29 (97%) acute thrombi, while all 6 chronic thrombi had a partial response. In contrast to the recommended adult-derived dosages of 0.1 to 0.5 mg/kg per hour, the authors found that initial doses of less than 0.01 mg/kg per hour were effective in 12 of 17 patients with acute thrombosis. Neonates required 0.06 mg/kg per hour. Route of administration (local or systemic) did not affect efficacy. Major bleeding occurred in only one extremely preterm infant. Minor bleeding, primarily oozing at intravenous sites, occurred in 27% of children during TPA infusions. Prophylactic unfractionated or low-molecular-weight heparin was infused concomitant with TPA in 42% of the children and did not increase the risk of bleeding. CONCLUSIONS: TPA in very low doses appears to be safe and effective for thrombolysis of acute thromboses in most children, given appropriate patient selection. PMID- 12759625 TI - Subclinical parvovirus B19 infection in children with sickle cell anemia. AB - PURPOSE: To investigate the prevalence and clinical consequences of previous parvovirus B19 exposure in a large cohort of pediatric patients with sickle cell anemia (SCA). METHODS: Prospective serologic testing for previous parvovirus B19 exposure was performed in steady-state pediatric patients with SCA, either prior to starting hydroxyurea therapy or in preparation for transition to the adult service. A retrospective chart review was performed to ascertain whether patients had a documented history of a transient aplastic crisis. RESULTS: The prevalence of serologic evidence of previous parvovirus infection increased with age. The overall prevalence in 102 children with SCA was 53%, ranging from 44% between 5 and 9 years of age to 71% between 17 and 21 years of age. Only 27% of patients had a previous clinically recognized transient aplastic crisis. CONCLUSIONS: By the teenage years, most pediatric patients with SCA have serologic evidence of previous parvovirus B19 exposure. However, subclinical parvovirus infection appears to be common in children with SCA, since most patients have no documented previous transient aplastic crisis. PMID- 12759626 TI - Positive blood cultures in sickle cell disease: time to positivity and clinical outcome. AB - PURPOSE: To prospectively identify all cases of bacteremia in children with sickle cell disease (SCD), establish time to positivity for various microorganisms, correlate clinical findings with microbiology data, and determine the antibiotic resistance pattern of the pneumococcal isolates. METHODS: All positive blood cultures from children with SCD followed at the Children's Hospital of Philadelphia from January 1993 through May 2001 were included. Isolates were classified as pathogen or contaminant. Demographic and clinical information was abstracted from the medical records. Time to positivity and antibiotic resistance data were generated in the microbiology laboratory. RESULTS: One hundred forty-one positive blood culture bottles were obtained during distinct febrile episodes. Thirty-nine percent contained pathogens and 61% contained contaminants. The average time to positivity was 17.1 hours in the pathogen group and 29.5 hours in the contaminant group (P < 0.0001). Streptococcus pneumoniae was the most common pathogen (42% of total), with a mean patient age of 3.5 years. Gram-negative rods were the second most common organism (28% of total), with a mean patient age of 8.1 years. Thirty-one percent of the pneumococcal isolates were resistant to penicillin. Thirty-five percent of the pneumococcal isolates grew from children with a focus of infection. Acute chest syndrome was noted in 26% of patients with a positive blood culture for S. pneumoniae. Sixty-seven percent of Salmonella isolates and 50% of Staphylococcus aureus isolates grew from patients who developed osteomyelitis. CONCLUSIONS: The average time to positivity for pathogens can be used in conjunction with other factors to determine the length of observation required for children with SCD who present with febrile illness. Chest radiographs should be obtained on children with SCD who are bacteremic with S. pneumoniae. Bone scans should be obtained on children with SCD who are bacteremic with Salmonella or S. aureus. PMID- 12759628 TI - Interaction of all-trans-retinoic acid with fluconazole in acute promyelocytic leukemia. AB - All-trans-retinoic acid (ATRA) has a dramatic antitumor effect in patients with acute promyelocytic leukemia (APL). It is hepatically metabolized by cytochrome P 450, and there are known toxicities associated with high levels of this drug. The effects of ATRA can be potentiated by inhibition of cytochrome P-450, which is known to occur with certain drugs. We report a case of a patient with ATRA toxicity thought to be secondary to interaction with fluconazole. PMID- 12759627 TI - Novel translocation in acute megakaryoblastic leukemia (AML-M7). AB - The authors report a unique translocation in a patient with M7 acute myeloid leukemia and review the literature. A 22-month-old girl without Down syndrome was diagnosed with acute myeloid leukemia, subtype M7 (AML-M7), and died with relapsed disease following bone marrow transplantation. Tumor cells were evaluated using cytogenetics (including spectral karyotyping), immunohistochemistry, and flow cytometry. The patient was found to have a previously unreported complex translocation as follows: 50,XX,der(1)t(1;5)(p36?.1;p15?.1),del(5)(p15?.1), +6,+der(6;7)(?;?),der(7)t(6;7)(?;p22)[2],der(9)t(6;9) (?;p21)t(9;14)(q34;q11.2 q13),+10,t(12;16)(p13;q24),-14[2], del(14)(q13)[2],+der(19)t(1;19)(?;p13.3),+22[cp 4]. AML-M7 in non-Down syndrome patients is a rare disease that requires improved prognostic markers. PMID- 12759629 TI - Pulmonary veno-occlusive disease after autologous bone marrow transplant in a child with stage IV neuroblastoma: case report and literature review. AB - Pulmonary veno-occlusive disease (PVOD) is a rare, almost universally fatal complication of chemotherapy and bone marrow transplantation with few treatment options. A 19-month-old boy with stage 4 neuroblastoma with fatal PVOD following high-dose chemotherapy with autologous peripheral blood stem cell rescue is described here. A comprehensive literature review revealed 40 case reports of PVOD in oncology patients. Various therapeutic modalities were attempted, with four survivors. PVOD should be considered in patients with dyspnea and cardiomegaly. Less invasive diagnostic methods and more effective therapies are needed. PMID- 12759630 TI - An adolescent with ovarian osteosarcoma arising in a cystic teratoma. AB - A 14-year-old girl had an abdominal mass with the characteristics of an ovarian germ cell tumor on computed tomography scan. The mass, arising from the left ovary, was completely resected and found to be osteosarcoma arising from a mature cystic teratoma. A metastatic lesion in the abdomen did not respond to 2 courses of cisplatin, doxorubicin, ifosfamide, and high-dose methotrexate, and was resected. Seven months after completion of chemotherapy, there were simultaneous local recurrence and lung metastases. Previously, 10 cases of ovarian osteosarcoma have been reported in the literature: 5 were primary osteosarcoma of the ovary, 4 were associated with teratomas, and 1 was part of a malignant mixed mesodermal tumor of the ovary. Of the 10, there are only 2 long-term survivors, both of whom were treated with adjuvant chemotherapy following complete resection. PMID- 12759631 TI - Mechanism of transient adrenal insufficiency with megestrol acetate treatment of cachexia in children with cancer. AB - Megestrol acetate (MA) is used to stimulate appetite in children with cancer, but it can impair adrenal function and cause adrenal insufficiency. Three children with brain tumors were treated with MA to improve weight gain. Standard low-dose ACTH stimulation tests were performed on and off MA. All patients failed the low dose ACTH stimulation while on MA and responded normally once off MA. The suppression of the adrenal axis was at the level of the hypothalamus/pituitary. Adrenal suppression can occur in patients treated with MA. Awareness of this possibility and the use of glucocorticoid stress coverage are advised. PMID- 12759632 TI - Prenatal exclusion of severe factor VII deficiency. AB - A nonconsanguineous asymptomatic couple, were identified as carriers of factor VII (FVII) deficiency when two of their newborn children died of massive intracranial hemorrhage secondary to severe congenital FVII deficiency. Complete sequence analysis of the factor VII (F7) gene in this couple indicated that the mother was heterozygous for an A to G transition at position -2 of the exon 5 acceptor splice site, and the father was heterozygous for a G to T transversion at position +1 of the exon 6 donor splice site. This information allowed us to exclude a compound heterozygous deficiency state in a subsequent pregnancy using PCR/direct sequencing of the F7 gene using DNA obtained from chorionic villi at 10 weeks' gestation. Our experience with the family reported here further supports the conclusion that mutation-specific detection is reliable in the prenatal exclusion of severe bleeding disorders. PMID- 12759633 TI - Use of vincristine and cyclosporine in childhood thrombotic thrombocytopenic purpura. AB - Thrombotic thrombocytopenic purpura (TTP) is a rare but life-threatening hematologic disorder in children. Plasmapheresis, the standard therapy for TTP, is effective in achieving remission in most patients. However, some patients become either refractory to or dependent upon plasmapheresis. The authors report three such patients in whom the use of vincristine or vincristine plus cyclosporine resulted in permanent remission. A 12-year-old girl with TTP dependent on plasmapheresis for more than 5 months responded to vincristine with a decrease in the required frequency of plasmapheresis, but the addition of cyclosporine abrogated the need for further plasmapheresis. She subsequently developed serologic evidence of systemic lupus erythematosus. Two 15-year-old boys with TTP (one of them with underlying mixed connective tissue disease) became refractory to plasmapheresis after a brief initial response. The addition of vincristine in one patient and vincristine and cyclosporine in the second (with mixed connective tissue disease) led to complete remission. The authors' experience in this case study of three patients suggests that vincristine and cyclosporine are effective agents in the management of patients with TTP who do not achieve complete remission with plasmapheresis alone. PMID- 12759634 TI - The CCC system: is it really the answer to pediatric MDS? PMID- 12759635 TI - Two-part and three-part fractures of the proximal humerus treated with suture fixation. AB - OBJECTIVE: To evaluate the radiographic and clinical outcomes of patients with displaced proximal humerus fractures (two-part and three-part) treated with nonabsorbable rotator cuff-incorporating sutures. DESIGN: Retrospective. SETTING: University hospital. PATIENTS: There were 27 patients (28 shoulders) with displaced proximal humerus fractures. There were 13 greater tuberosity (GT) and 9 surgical neck (SN) two-part fractures and 6 GT/SN three-part fractures. The average age was 64 years (range 38 to 84 years). The average follow-up was 4.4 years (range 1.0 to 11.5 years). INTERVENTION: All patients were surgically treated solely with heavy polyester nonabsorbable sutures. MAIN OUTCOME MEASUREMENTS: Functional assessment was obtained using the American Shoulder and Elbow Surgeons (ASES) score and Neer's criteria, which grade outcomes as excellent, satisfactory, or unsatisfactory. RESULTS: Overall, there were 22 (78%) excellent, 3 (11%) satisfactory, and 3 (11%) unsatisfactory results, and the average ASES score was 87.1 (range 35.0 to 100.0). All shoulders healed radiographically without evidence of avascular necrosis of the humeral head. Twenty-four shoulders (86%) had anatomic alignment on postoperative radiographs. Of four shoulders with nonanatomic alignment, three had ASES scores of >/=90, with excellent Neer scores. When comparing patients with isolated two-part GT fractures (n = 13) with patients having two-part SN or three-part SN/GT fractures (n = 15), there were no statistically significant differences with respect to range of motion (P > 0.05) and outcome measures (P > 0.05). All patients who had unsatisfactory outcomes were noncompliant with physical therapy, with ASES scores averaging 39.4 (range 35.0 to 43.3). CONCLUSION: Two-part and three-part GT and SN fractures can be treated satisfactorily with heavy nonabsorbable rotator cuff incorporating sutures, particularly in elderly patients. Hardware-associated complications are obviated. Patients with SN fractures treated with sutures can have outcomes similar to patients with two-part GT fractures. Although the goal is to reconstruct a "one-part" fracture pattern, some residual deformity does not preclude an excellent outcome. A compliant patient is crucial for a successful result. PMID- 12759636 TI - Sliding osteotomy for deformity correction following malunion of volarly displaced distal radial fractures. AB - OBJECTIVE: To determine the outcome after a sliding osteotomy for deformity correction following malunion of volarly displaced distal radius fractures. DESIGN: Retrospective review of a consecutive patient series. SETTING: A university-affiliated, tertiary-care center. PATIENTS/INTERVENTION: Ten patients with symptomatic distal radius malunion following a volarly displaced distal radial fracture (Smith's fracture) were treated with an oblique sliding osteotomy and plate fixation, through a volar approach, without using an iliac crest bone graft. Five men and five women with an average age of 41.9 years were followed for an average of 2.7 years postoperatively. MAIN OUTCOME MEASUREMENTS: Range of motion, grip strength, Fernandez wrist score, radiographic parameters. RESULTS: At latest follow-up, wrist extension improved from an average of 37 degrees preoperatively to 70 degrees postoperatively (P = 0.002), wrist flexion improved from an average of 40 degrees to 65 degrees (P = 0.012), and supination improved from an average of 31 degrees to 68 degrees (P = 0.002). Postoperative radiographs revealed an average deformity correction of 10.6 degrees of volar tilt, 7.7 degrees of radial inclination, 5.8 mm of ulnar variance, and 10.4 mm of volar translation. Using the Fernandez point score (0-20) system, the average overall score improved from 10.5 preoperatively to 17.6 postoperatively (P = 0.0001). Functional outcome was rated as excellent or good in 9 of 10 patients and fair in 1 patient (who experienced residual problems due to persistent ulnar sided pain). There were two reoperations (one hardware removal, one distal ulnar hemiresection). CONCLUSIONS: This method reliably restores distal radial anatomy, decreases pain, and improves supination without requiring iliac crest bone grafting. PMID- 12759637 TI - Femoral neck stress fractures: outcome analysis at minimum five-year follow-up. AB - INTRODUCTION: The complications associated with misdiagnosed or undertreated femoral neck stress fractures in young, active adults have been well documented in the orthopaedic literature. Less is known regarding the outcome of these injuries in patients whose diagnosis was timely and whose treatment was appropriate. METHODS: A sample of 25 patients previously involved in an unrelated study evaluating femoral neck stress fractures were contacted retrospectively 5 to 7 years after their injury. They were asked to complete a self-administered outcome evaluation, the Musculoskeletal Function Assessment (MFA), and answer several specific questions regarding their hips at the present time. Their MFA score was compared with treatment method, fracture type, and bone mineral density (BMD). RESULTS: All 25 patients responded to our inquiries. Of patients, 68% continued to feel "somewhat bothered" by their injury in at least one functional category. Nine patients felt "disabled." No patient has developed avascular necrosis, nonunion, malunion, or posttraumatic arthrosis or was currently under the care of an orthopaedic surgeon. Nine patients had developed stress fractures in other locations. The mean MFA score was 18.80 (range 0 to 63). A lower score corresponds to a patient's perceived higher level of function. Analysis of MFA scores did not reflect statistically significant differences between fracture location, treatment modality, or BMD. CONCLUSIONS: Femoral neck stress fractures can result in devastating problems for young adults. Appropriately treated patients, regardless of treatment method, may have persistent complaints. PMID- 12759638 TI - Heterotopic ossification after knee dislocation: the predictive value of the injury severity score. AB - OBJECTIVE: To determine the relationship of multiple variables, including the Injury Severity Score (ISS), closed head injury (CHI), and timing and type of surgery to formation of motion-limiting heterotopic ossification (HO) following knee dislocation. DESIGN: Longitudinal observational study. SETTING: University level 1 trauma center. PATIENTS/PARTICIPANTS: Thirty-five consecutive patients with 36 knee dislocations (OTA fracture and dislocation classification 40-D) admitted over a 26-month period. MAIN OUTCOME MEASUREMENTS: Admission ISS, Glasgow Coma scale (GCS) scores, CHI, timing (> or < 3 weeks from injury) and type (open or arthroscopic) of surgery, number of cruciate ligaments reconstructed, medial surgical procedure, and eventual presence or absence of motion-limiting HO. RESULTS: A classification system for HO was developed ranging from none (type 0) to ankylosing (type IV) HO. Twenty-nine patients with type 0 III HO recovered an average range of motion of 126 degrees at an average of 14 months (group A). Six patients formed ankylosing type IV HO (group B). The ISS in group A ranged from 9 to 26. ISS in group B ranged from 26 to 50 (P < 0.001). Regarding the formation of type IV HO, the sensitivity of an ISS >/=26 was 100%, the specificity was 97%, and the positive predictive value was 86%. Patients in group B had a greater incidence of documented CHI (P < 0.025). Timing and type of surgery, number of ligaments reconstructed, and whether or not the patient had a medial surgical procedure had no statistical influence on degree of HO formation. CONCLUSIONS: An ISS of 26 seems to be a discrete boundary above which patients with knee dislocation are at extremely high risk for type IV HO formation if undergoing surgical reconstruction and below which patients are likely spared this complication. The presence of a CHI is a significant factor in type IV HO formation, although harder to quantify. None of the remaining independent variables studied were significantly related to ankylosing type IV HO formation. PMID- 12759639 TI - Treatment of displaced bicondylar tibial plateau fractures (OTA-41C2&3) in patients older than 60 years of age. AB - OBJECTIVE: To investigate the outcome (clinical, radiographic, and general health status) of the surgical treatment of displaced bicondylar tibial plateau fractures (OTA-41C) in patients >60 years old. DESIGN: Prospective cohort study. SETTING: Limb reconstruction service, university teaching hospital. PATIENTS: Eleven consecutive patients >60 years old with a mean age of 72 years (range 60 to 90 years). The indications for surgery were displaced bicondylar tibial fractures, open fractures, and fractures with severe soft tissue injury. INTERVENTION: All patients were treated according to a standard protocol, which involved limited articular reconstruction and percutaneous intrafragmentary screw fixation, followed by neutralization with a stable beam-loading external fixator and early mobilization. MAIN OUTCOME MEASUREMENTS: The clinical outcome was assessed using Rasmussen's system and the Iowa knee score. For general health assessment, the anglicized SF-36 was used. Radiographic assessment was performed for redisplacement and angulation on digitized radiographs. RESULTS: After a mean follow-up of 38 months (range 18 to 51 months), bony union was achieved in all patients. Seven of 11 patients started full weight bearing 2 to 6 weeks postoperatively. According to Rasmussen's system, 9 of 11 (82%) scored satisfactory results. Radiographic redisplacement was found in three severely comminuted cases resulting in >/=10 degrees of valgus malunion. One patient received a corrective osteotomy while still in the fixator. Another needed TKA. Limited knee flexion was found in three patients with cross knee fixation. Superficial pin site infection occurred in five patients, but there were no cases of deep infections or septic arthritis. CONCLUSIONS: All-ring external fixation, as a beam-loading system applied in a neutralizing mode, is a safe, stable, and reliable technique for the treatment of displaced bicondylar tibial plateau fractures in elderly patients. PMID- 12759640 TI - Predictors of reoperation following operative management of fractures of the tibial shaft. AB - BACKGROUND: Accurate prediction of likelihood of reoperation in patients with tibial shaft fractures would facilitate optimal management. Previous studies were limited by small sample sizes and noncomprehensive examination of possible risk factors. OBJECTIVE: We conducted an observational study to determine which prognostic factors were associated with an increased risk of reoperation following operative treatment in a heterogeneous population of patients with tibial shaft fractures. DESIGN: Retrospective observational study. SETTING: Level 1 trauma center. METHODS: We identified 200 patients with tibial shaft fractures from two university-affiliated centers. Two reviewers independently abstracted data regarding 20 possible prognostic variables, reviewed preoperative and postoperative radiographs, and documented reoperations (defined as any surgical procedure 35 years (P=0.06). The clinical course of pancreatitis (severity and complication rates) was not altered by the presence of a mutation. CONCLUSION: One-quarter of all patients and one-third of those < or =35 years with idiopathic pancreatitis have at least one mutation of the CFTR gene. The presence of a CFTR mutation appears to predict the development of pancreatitis at an earlier age. PMID- 12759681 TI - [Are the causes similar for benign and severe forms of acute pancreatitis?]. AB - The frequency of severe acute pancreatitis not due to alcohol or biliary causes is not well known. AIMS: To evaluate the distribution of causes responsible for benign and severe cases of acute pancreatitis in an effort to identify causes to search for in patients with severe acute pancreatitis. PATIENT: All patients hospitalized for acute pancreatitis between January 1994 and May 2001 with a good quality CT scan. METHODS: All patients had a complete, standardized evaluation to look for all possible causes of acute pancreatitis. The following severity criteria were retrospectively reviewed: maximal C-reactive protein level, Ranson's score, Balthazar's score, percentage of patients hospitalized in intensive care unit or a high-dependency unit, hospitalization duration, and local or general complications. RESULTS: One hundred thirty-nine patients were included. The cause of acute pancreatitis were: alcohol (34%), biliary (27%), obstructive (16%), miscellaneous (10%), unknown (9%), post endoscopic retrograde cholangio-pancreatography (4%). The studied severity factors did not differ with respect to the cause of acute pancreatitis with the exception of Balthazar's score. Non-alcoholic non-biliary causes were found in 19 (27%) of the 71 patients with severe necrotic acute pancreatitis (Balthazar > or =D) and 35 (51%) of the 68 patients with acute pancreatitis with Balthazar score or =D). For the other severity scores, the distribution of causes was similar. After exclusion of biliary and alcoholic causes, a careful search for other etiologies should be carried out in both benign and severe cases of acute pancreatitis. PMID- 12759682 TI - [Pseudomyxoma peritonei treated with complete resection and immediate intraperitoneal chemotherapy]. AB - AIM: Pseudomyxoma peritonei remains a fatal disease. This clinical pathological entity based on the presence of mucin includes different prognostic groups. Complete resection of macroscopic lesions, combined with immediate intraperitoneal chemotherapy to treat remnant infra-millimetric disease, might improve survival. The aim of this prospective study was to evaluate this treatment strategy. METHODS: Thirty-six patients with pseudomyxoma peritonei underwent resection of supra-millimetric lesions then were given either early postoperative intraperitoneal chemotherapy (5 days) (before January 1996) or intraoperative chemohyperthermia treatment (after January 1996). During this same period, only partial resection of the macroscopic lesion was possible in 15 patients; these patients were not given peritoneal chemotherapy. RESULTS: Postoperative mortality was 13.8% (n=5), including 2 deaths not specifically due to the procedure. Morbidity, including severe and non-severe complications was 44%. After a mean follow-up of 48 months, the overall 5-years survival rate was 66%, and disease-free survival rate was 55% (including the postoperative deaths). The main prognostic factor in this series was the pathological grading: 5-years survival was 74% for grade 1 tumors versus 54% for grades 2-3 (P=0.05). CONCLUSION: The main prognostic factor of the pseudomyxoma peritonei, after the completeness of the resection, is the pathological grading. The addition of an intraperitoneal chemohyperthermia improves long-term survival of grades 2-3 tumors and perhaps that of grade 1 (agreement of experts). This treatment is more easily performed, more well-tolerated, and more efficient when performed early. PMID- 12759683 TI - [Cystadenoma of the liver with high levels of ACE and CA 19-9 in the cyst]. AB - The development of a cystadenocarcinoma from previously benign cystadenoma is controversial. Neither clinical nor biological factors have been described to explain this transformation. High levels of serum and cystic CEA and CA 19-9 seem to help in the diagnosis of cystadenoma but not cystadenocarcinoma. Definitive histological evaluation is the only means to discriminate malignant from benign cysts. We report a case of cystadenoma of the liver with very high cystic levels and normal serum levels of CEA and CA 19-9. PMID- 12759685 TI - [Quinine-induced hemolytic-uremic syndrome in a patient with cirrhosis]. PMID- 12759684 TI - [Endoscopic and pathological aspect of small bowel and colonic involvement in Whipple's disease: a case report]. AB - We describe a case of Whipple's disease confirmed by clinical, histological, bacteriological and molecular criteria. The duodenal involvement was associated with the presence of an endoscopic and histological enterocolitis. Final diagnosis of small bowel and colonic involvement by Whipple's disease was confirmed by histology and molecular biology. We review the literature on extra duodenal involvement. We underline the fact that enterocolitis in Whipple's disease is non-specific. We also discuss the other causes of intestinal mucosal infiltration by macrophages. PMID- 12759686 TI - [A new case of familial Caroli's disease]. PMID- 12759687 TI - [Aeroportia and necrotizing enteritis after chemotherapy with paclitaxel for lung carcinoma]. PMID- 12759688 TI - [Di-Antalvic-induced esophageal ulcer]. PMID- 12759689 TI - [Non-adenomatous colorectal polyps may warrant surgical treatment]. PMID- 12759690 TI - [Medical treatment of a voluminous tumor of the mesenteric root]. PMID- 12759691 TI - [Response to hepatitis A and hepatitis B vaccination: genetic differences]. PMID- 12759692 TI - Management of variceal hemorrhage. PMID- 12759694 TI - Abstracts of the 17th European Histocompatibility Conference and 11th Annual Meeting of the German Society of Immunogenetics. 6-9 May 2003, Baden-Baden, Germany. AB - Genes and Immunity (2003) 4, S1-S4. doi:10.1038/sj.gene.6363987 PMID- 12759693 TI - Organ donation and treatment of the multi-organ donor. PMID- 12759707 TI - [Smoking prevention, breathable air protection and health promotion in relation to smoking: advancing towards a new future]. PMID- 12759708 TI - [Scientific evidence based medicine: myth and reality of variability in clinical practice and its impact on health outcomes]. AB - Clinical practice is not an exact and reproducible phenomenon since it is carried out in a working framework with a high degree of uncertainty. The different sources of variability in clinical practice are analysed, with special emphasis placed on the quality of the scientific evidence that underlies decision making. On the basis of the change from the traditional model to a practice based on information derived from scientific evidence, different problems are examined in relation to the need for transforming the information into effective action. Finally, with the aim of reducing variability and its harmful impact on health, those areas that should be the object of future analysis and research are enumerated. PMID- 12759709 TI - [Resistance to penicillin and other antimicrobials in 103 clinical isolations of Streptococcus pneumoniae (2000-2001)]. AB - BACKGROUND: To determine in our hospital the sensitivity of isolations of Streptococcus pneumoniae to penicillin, as well as to analyse the association of resistance to penicillin and other antimicrobials and the activity of cefotaxime and cefepime in pencillin resistant strains of Streptococcus pneumoniae. METHODS: The sensitivity was determined on 103 isolations of Streptococcus pneumoniae, from clinical samples from the years 2000-2001, to penicillin, eritromycine, cloramfenicol, tetracycline, cotrimoxazol, cefotaxime, cefepime and levofloxacine. RESULTS: Sixty-eight percent of the isolations were sensitive to penicillin, while some 32% of the isolated strains of Streptococcus pneumoniae were penicillin resistant, with 7.7% showing a high degree of resistance. Resistance to eritromycine, cloramfenicol, tetracycline, cotrimoxazol and levofloxacine was 38.8%; 9.7%; 20.4%; 25.2% and 2.9% respectively, increasing to values of 66.6%; 30.3%; 48.5%; 72.7% and 9.1% in the 33 penicillin resistant strains. Resistance to cefotaxime and cefepime was 9.7% and 10.6% respectively. CONCLUSIONS. A high percentage of strains of Streptococcus pneumoniae show some degree of resistance to penicillin, but with lower figures than in other studies carried out at a national level. Similarly, it was demonstrated that resistance to penicillin is significantly associated (p < 0.001) with resistance to all the antimicrobials studied, except for levofloxacine. The resistances to cefotaxime and cefepime were comparable, with less activity being observed in these when facing penicillin resistant strains of Streptococcus pneumoniae. PMID- 12759710 TI - [Evaluation of the clinical practice of Informed Consent in clinical trials]. AB - On the basis of existing publications it would seem legitimate to assume that in a clinical test (CT) the difficulties inherent in the process of researcher participant communication are in practice greater than desired. Similarly, the hypothesis is adopted that difficulties exist in the formal legibility of the documents of Informed Consent. We present the results of a transversal study made of a random sample (n=160) of the CTs approved by the Ethical Committee of Clinical Research (CEIC) of Navarra during the years 1995-1999. The results found were: in 69.7% of the cases the researcher filed the documents corresponding to the CT, the Informed Consent appears signed by the researcher in 56.6% of the CTs, and in more than 83% of the cases the written information shows shortcomings in legibility, which confirms the correctness of the hypothesis and permits the detection of areas where improvements need to be developed. PMID- 12759711 TI - [Etiopathogenesis of giant cell arteritis]. AB - Giant cell arteritis is a vasculitis of large and medium size arteries, especially those of the aortic arch with an extracranial distribution, but also the aorta and some of its larger branches. It is characterised by the presence of mononuclear inflammatory infiltrates close to the internal elastic lamina formed by lymphocytes and macrophages, which in slightly more than 50% of the cases contain multinucleate giant cells. The morbidity associated with this disease is related to phenomena of distal ischemia to the luminal stenosis of the inflamed arteries, and to a lesser extent to the formation of aneurisms due to the weakening of the arterial wall. With an unknown aetiology, its pathogenesis is immune through the migration and location of gamma-INF -producing T cells in the adventitia of the inflamed arteries, it being assumed that this is the place of immune stimulation by a still unidentified antigen. The recruitment and activation of macrophages by this cytokine is one of the most important points of its pathogenesis. The destruction by these of the arterial elastic tissue is a relevant phenomenon, as is the production of other factors promoting neoangiogenesis and a proliferation of neointime, responsible through obliterating light for the ischemic manifestations of the disease. The process is accompanied by an important systemic repercussion characterised by a strong reaction of acute phase and general but barely specific symptoms of disease. On the other hand, an important percentage of patients show a clinical picture of polymyalgia rheumatica, an entity that has a historical and controversial relationship to this arteritis. In recent years important contributions have been made to the understanding of the immune mechanisms involved in its pathogenesis. PMID- 12759712 TI - [Updating Fanconi's anaemia]. AB - Fanconi's anaemia (FA) is an autosomal recessive syndrome associated with chromosomal instability, and hypersensitivity of the DNA to claustrogenic agents. Clinically it presents a progressive marrow insufficiency, different congenital anomalies and an predisposition to malignancy. Eight complementation groups have been defined and the genes corresponding to six of them have been cloned. Recent advances in molecular biology have made it possible to investigate the relationship between the FA genotype and the nature and severity of the clinical phenotype. The treatment of FA is also the object of intense research that is currently centred on the transplant of hematopoyetic progenitors, especially successful in cases of an HLA-identical brother or sister donor, and in gene therapy, which is still in the phase of clinical research. PMID- 12759713 TI - [Population-based epidemiology of colorectal cancer: causality review]. AB - The estimated number of new cases of colorectal cancer per year in Spain (no. 19,166) is higher than other tumour locations. 1.56 times more cases of colon cancer are registered than of the rectum, and there are 1.44 times more cases in men than in women. Incidence and mortality are lower than the average for European countries; in historical series (1973-1999) an increase can be observed by age, period and birth cohorts between 1898 and 1932. On the contrary, in the USA a reduction of mortality can be observed from 1973 to 1999 (-20.8%) and of incidence from 1985 to 1999 (-7.4%). In Spain, the average duration of the disease in years is 4.29, lower than that of the European Community (4.57), and 72% of the figure for the most favourable country (5.93). Relative survival after five years in Spain is lower than in the USA (61.9% vs. 54%). Several dietary, environmental and lifestyle factors appear to be associated with colorectal cancer, but the risk or protection of these factors are of little weight and the results of studies are at times contradictory. The reduction of incidence in the USA indicates that intervention is possible to bring about a change of trend, predictably by means of secondary prevention. PMID- 12759715 TI - [Protection of breathable air: the new Law on smoking prevention in Navarra (2003)]. PMID- 12759714 TI - [Diseases of Compulsory Notification (DCN) in Navarra. 2002]. AB - Since 1998, the Epidemiological Survelliance System of Navarra has included the notification of 34 transmissible infectious diseases, to which are added epidemic outbreaks of any aetiology and cause. Reporting to the system is carried out on a weekly basis by every doctor who suspects, or diagnoses, any of these processes. In our autonomous community, Diseases of Compulsory Notification (DCN) are reported using standardised forms on a weekly basis to the Section of Infectious Diseases and Control of Outbreaks of the Public Health Institute. Notification is made by the doctors and/or paediatricians of Primary Care and by certain services of Specialised Care. Subsequently, the information is sent to the National Epidemiology Centre where data from the Autonomous Communities is centralised and diffused. The year 2002 marks the fifth year of the new Epidemiological Vigilance System. In these five years there have been 74,814 notifications of disease, 17,184 in the year 2002, which provides a balance of notification of 74.07% for this year. In 2002, under the heading of respiratory transmitted diseases, 24,870 cases of Inluenza were reported, Epidemic Index (EI: 0.80). 58% of total annual cases were reported in the first nine weeks of the year, with a maximum in week 4 when 3,277 cases were reported. 16 cases of Meningococcal Disease were reported to the system (EI: 1.07). All the cases were confirmed microbiologically and all appeared in a sporadic way. With respect to the causative serogroup, on 12 occasions Neisseria meningitidis serogroup B was isolated and in the 4 remaining cases serogroup C was isolated. One case was notified in infants of less than 2 years of age (Rate: 10.52 cases per 100,000), another case in children between 2 and 5 years (5.52 cases per 100,000), 5 cases in the age group of 6 to 19 years (Rate: 5.86 cases per 100,000) and the remaining nine cases in the age group of persons aged 20 years or over (2.2 per 100,000). 70 cases of Legionellosis were declared in 2002 (EI: 4.67), all but one under the clinical form of pneumonia. Twenty-two of the cases were presented in the context of two outbreaks with a community origin, which affected 17 and 5 persons respectively. Similarly, there was a notable increase in the declaration of cases of bacillary dysentery, with 6 cases (EI: 2.00), brucellosis, with 10 cases (EI: 1.67) and chickenpox, with 4,346 notified cases (EI: 1.61). PMID- 12759716 TI - [Sudden death in a normal heart. Idiopathic ventricular fibrillation. Review of the literature concerning one case]. AB - Idiopathic ventricular fibrillation is that which is produced in the absence of structural cardiac disease and of other identifiable causes of ventricular fibrillation such as cardiotoxicity, electrolytical alterations or hereditary predisposition. The case of a healthy male, aged 37, who was asymptomatic until the day he was admitted to hospital where he showed numerous episodes of ventricular fibrillation without any previous triggering, is discussed. In the examination no cause was found to explain this, and an automatic defibrillator was implanted. The requirements for its diagnosis, risk stratification and the usefulness of the tests employed, as well as the treatments proposed are discussed. PMID- 12759717 TI - Effects of atropine on human cardiac beta 1- and/or beta 2-adrenoceptor stimulation. AB - The aim of this study was to find out whether, in humans, the increase in vagal tone accompanying cardiac beta-adrenoceptor (beta-AR) stimulation might be different dependent on beta1- or beta2-AR stimulation. For this purpose we studied, in six male healthy volunteers (aged 28+/-1 years), the effects of atropine infusion (0.15 microg/kg/min continuously) on increase in heart rate (HR) and contractility (determined as shortening of HR-corrected duration of electromechanical systole-QS2c) evoked by infusion of isoprenaline (3.5-35 ng/kg/min, increasing HR and QS2c via beta1-and beta2-AR), terbutaline (25-150 ng/kg/min, increasing HR and QS2c via beta2-AR), adrenaline (20-160 ng/kg/min, increasing HR via beta2-and QS2c via beta1-AR) and bicycle exercise in supine position (increasing HR and QS2c via beta1-AR). The three beta-AR agonists and exercise increased HR and shortened QS2c in a dose- or work-load-dependent manner, respectively. Atropine enhanced HR-increasing effects of all three beta AR agonists and exercise; increases were larger for beta2-AR (terbutaline, adrenaline) mediated effects than for beta1-AR (exercise) mediated effects. Moreover, atropine enhanced beta-AR agonists-induced QS2c shortening; however, atropine effects on QS2c were markedly less pronounced than on HR. From the results we conclude that, in humans, beta1-and beta2-AR mediated stimulation evoked HR-increases are composed of two components: increases via direct beta-AR stimulation and simultaneously decreases via increase in vagal tone. In addition, beta-AR mediated increases in contractility are also dampened by simultaneous activation of vagal tone but to a lesser extent possibly because human ventricular myocardium is only sparsely parasympathetically innervated. PMID- 12759719 TI - Health professional's guide to rehabilitation of the patient with osteoporosis. PMID- 12759718 TI - Characterisation of [125I]-Tyr0DTrp8-somatostatin binding in sst1- to sst4- and SRIF-gene-invalidated mouse brain. AB - Five somatostatin receptors (sst) have been cloned and mRNAs for the first four (sst1-4) are expressed in many brain regions. In the present work, we compared the distribution of the non-selective ligand [125I]-Tyr0-DTrp8-SRIF14 by autoradiography in 24 brain regions and pituitary in wild type, sst1- to sst4- or SRIF-gene invalidated (KO) mice. [125I]-Tyr0-DTrp8-SRIF14 binding was not significantly modified in sst1 KO mouse brain with the noticeable exception of the substantia nigra and only moderately decreased in pituitary. For sst2 KO mice, a general decrease (>75%) was observed in most regions, with the noticeable exception of the olfactory bulb and CA1 field of the hippocampus. SST3 KO brain displayed a decrease in binding in the external plexiform layer of the olfactory bulb only (-54%). For sst4 KO mice, [125I]-Tyr0-DTrp8-SRIF14 binding levels in the external plexiform (-35%) and glomerular (-39%) layers of the olfactory bulb as well as the hippocampus CA1 field (-68%) were significantly decreased. In SRIF KO mice, a significant increase in binding levels was observed in olfactory bulb, anterior olfactory nucleus, frontal cortex upper layers, lateral septum, CA1 field, zona incerta and lateral hypothalamus, substantia nigra, periaqueductal grey and parabrachial nucleus. Competition with selective ligands (CH275, octreotide or L-779,976, L-796,778, L-803,087, and octreotide or L-817,778, for sst1-5 receptors, respectively) was in accordance with these findings. Moreover, octreotide was still able to compete on residual [125I]-Tyr0-DTrp8-SRIF14 binding sites in sst2 KO pituitary. It is concluded that most [125I]-Tyr0-DTrp8-SRIF14 binding sites in mouse brain and pituitary belong to the sst2 subtype but for the olfactory bulb (sst3 and sst4 receptors), the CA1 of the hippocampus (sst4 receptors) and the pituitary (sst5 and sst1 receptors) in which other subtypes are also expressed. The overall increase in [125I]-Tyr0-DTrp8-SRIF14 binding in SRIF KO mice indicates that SRIF receptors, mostly from the sst2 subtype, are regulated by the endogenous ligand(s). PMID- 12759720 TI - Weight changes after renal transplantation: a comparison between patients on 5-mg maintenance steroid therapy and those on steroid-free immunosuppressive therapy. AB - After renal transplantation (RTx), an increase in body weight (BW) is usually observed, in which corticosteroids may play an important role. However, the effects of a low maintenance dosage of corticosteroids on BW have not been studied longitudinally in RTx patients. The aim of this study was to compare changes in BW after RTx in patients on steroid- or steroid-free immunosuppressive therapy and to assess the relationship between post-transplant weight changes and other potentially important factors. The charts of 123 RTx patients (72 male, 51 female) were retrospectively examined for BW changes in the first 5 years after RTx. Sixty-six patients were on 5-mg maintenance steroid dose and 57 patients underwent steroid-free immunosuppression. Mean post-transplant BW gain was 3.0+/ 5.3 kg after 6 months, 3.9+/-6.2 kg after 1 year and 6.2+/-8.6 kg after 5 years. Weight gain in the first year after RTx was related neither to maintenance- nor to cumulative steroid dose, age, gender, occurrence of rejection, or renal function. Weight gain was, however, significantly related to pre-transplant BMI and dialysis modality. After the first year, weight gain was significantly and positively related only to the cumulative steroid dose. The course of weight gain in the first year after RTx turned out to be independent from factors such as maintenance- or cumulative steroid dose, age, gender, occurrence of rejection, and renal function; weight gain was, however, dependent on pre-transplant BMI and dialysis modality. After the first year, the weight course was significantly affected by cumulative steroid dose. PMID- 12759721 TI - Kidney warming during transplantation. AB - The objective of the present study was to describe kidney temperature variations during transplantation and to identify the factors responsible for kidney warming. Kidney temperature was recorded steadily during transplantation. Kidney weight, body mass index (BMI), second warm ischemia time (t), and kidney temperature at the time of being placed in the recipient were analyzed so that we could evaluate their influence on kidney temperature and on kidney warming during transplantation. Kidney temperature at the time of removal from the container was 1.6 degrees C and 6. 3 degrees C when the kidney was placed in the recipient. Kidney temperature in the recipient depended on kidney temperature after serum washing (P<0.0001), but was independent of kidney preparation time (P=0.94). Then, kidney temperature (T) increased according to the logarithmic curve given in the following equation: T=7.2 ln(t)-0.6. Kidney temperature at the end of anastomosis was 26.7 degrees C. Kidney warming speed was 0.48 degrees/min and was dependent on the length of time of vascular anastomosis (P<0.0001). Kidney weight decreased the kidney warming speed (P=0.02). In conclusion, kidney warming increases slowly during ex vivo preparation. Kidney temperature stays below the damaging ischemic temperature of 18 degrees C. Because of its major impact on kidney warming, it is desirable that vascular anastomosis time be reduced, and, consequently, ex vivo kidney preparation needs to be meticulous. PMID- 12759722 TI - Long-term follow-up of ACE-inhibitor versus beta-blocker treatment and their effects on blood pressure and kidney function in renal transplant recipients. AB - Hypertension and nephrotoxicity are frequent complications of cyclosporine induced immunosuppression in renal transplant recipients. Long-term antihypertensive treatment is obligatory for hypertensive transplant patients, to protect allograft function. The use of angiotensin-converting enzyme (ACE) inhibitors in the anti-hypertensive treatment of renal transplant recipients who receive immunosuppression with cyclosporine has long been discussed controversially. The aim of this prospective study, with a duration of 2 years and a follow-up of another 3 years, was to estimate the long-term antihypertensive potential of quinapril compared with that of the beta-blocker atenolol and to compare their effects on renal allograft function and proteinuria in 96 hypertensive renal transplant recipients who received cyclosporine A as immunosuppressive therapy. Patients were randomly assigned to receive either quinapril (group Q) or atenolol (group A) as anti-hypertensive treatment. Forty patients of each group completed the 5-year observation period according to protocol. Intention-to-treat and according-to-protocol analyses were performed. With the patients starting at similar baseline blood pressure values, both agents, atenolol and quinapril, decreased systolic and diastolic blood pressure (SBP, DBP) as well as middle arterial pressure (MAP) and pulse pressure (PP) to a similar extent (Delta SBP: group Q: -8+/-3 vs group A mmHg: -5+/-3; Delta DBP: 5+/-2 vs -4+/-2 mmHg; Delta MAP: -6+/-2 vs -5+/-2 mmHg; Delta PP: -2+/-2 vs -1+/ 3 mmHg; mean +/- SEM). Neither serum creatinine levels nor Cockcroft-Gault clearance had changed significantly in either group after the 5-year period (Delta creatinine: 0.1+/-0.1 vs 0.2+/-0.2 mg/dl; Delta Cockcroft-Gault clearance: 3.9+/-4.6 vs 2.8+/-4.3 ml/min; mean +/- SEM). Urinary protein excretion remained stable among the quinapril-treated patients, whereas a significant increase was observed in the atenolol group during the 5-year study period (group Q: from 0.52+/-0.08 to 0.54+/-0.14 g/24 h; group A: from 0.34+/-0.03 to 0.72+/-0.13 g/24 h, P<0.02; mean +/- SEM). Albuminuria increased comparably in both groups, while the excretion of alpha-microglobuline increased slightly in the atenolol group, but decreased slightly in the quinapril group. The difference between the groups failed to be statistically significant (ANOVA, P<0.056). In conclusion, quinapril and atenolol may be considered suitable and safe substances in the long-term treatment of hypertensive renal transplant recipients, since both agents prove to be effective in anti-hypertensive treatment, and keep allograft function stable over a period of 5 years. PMID- 12759723 TI - Cost-benefit approach in evaluating investment into donor action: the German case. AB - Donor Action (DA) is an international initiative to help critical care units (CCUs) increase their own donation rates through improved-quality donation practices. Following a validated diagnostic review (DR), areas of weakness can be identified, and the appropriate changes introduced. Data gathered from a number of centers in nine European countries (including Germany) 1 year after the introduction of targeted improvement measures demonstrated a 59.2% (P=0.0015) increase in donation rates. This analysis computes the cost-benefit thresholds of implementing the DA methodology from a German health-economic point of view, taking into account the treatment alternatives for end-stage renal disease (dialysis and transplantation) and comparing the DA program with current organ donation practice. Lifetime direct medical costs and quality-adjusted life years (QALYs) were calculated for both arms, considering only changes in cadaveric renal transplantation rates. If DA leads to a 59% overall increase in organ donation in Germany, the program will result in 33 QALYs and 1.8-million euro cost savings per million population (PMP). Therefore, DA would be cost-effective below 2.66-million euro implementation cost PMP (or 218-million euro for the whole country). As the partial implementation cost of the program was far below the threshold, DA is more cost-effective than other publicly reimbursed medical intervention. PMID- 12759724 TI - The influence of surgery, immunosuppressive drugs, and rejection, on graft function after small bowel transplantation: a large-animal study. AB - In this study we assessed functional changes (motility and absorption) of intestinal allografts in a large-animal model of orthotopic small bowel transplantation in swine. Studies were performed on non-rejecting animals in the early and late stages after transplantation and after induction of different grades of acute rejection. Immunosuppression consisted of oral FK506 and mycophenolate mofetil. In each study group we regulated drug administration, in terms of dosage and timing, in order to induce different grades of acute rejection or to prevent it. Migrating myoelectrical complexes were recorded in fasting animals so that motility could be assessed. Mucosal biopsy of the allograft and D-xylose absorption tests were performed on the same day as the motility study. In the early stages following intestinal transplantation, we observed in non-rejecting animals a slightly increased graft motility and a marked carbohydrate malabsorption. Recovery of the carbohydrate absorption capacity occurs within 2 months, but the persistence of diarrhea leads to partial malabsorption and to a lack of normal weight gain. Motility reduction correlates with the grade of acute rejection and becomes significant at a later stage, when rejection is severe. Allograft carbohydrate absorption, on the contrary, is markedly reduced in all rejecting pigs, irrespective of the grade of rejection. In summary, the early functional impairment of non-rejecting animals has multifactorial causes due to surgery and immunosuppression (drug toxicity), and its occurrence suggests the need for specific guidelines for clinical early postoperative enteral feeding. The functional studies adopted here are helpful in defining the grade of functional impairment with or without acute rejection; however, they are not useful for early detection of ongoing acute rejection of the small bowel graft. PMID- 12759725 TI - Longitudinal study of cryptococcosis in adult solid-organ transplant recipients. AB - While studies in kidney recipients have found meningitis to be the most common clinical manifestation of cryptococcosis (Cry), it is unclear if the clinical presentation of Cry differs among various solid-organ transplant (SOT) recipients and whether the serum cryptococcal antigen (SCA) might predict the site of infection. We report the clinical manifestations and the correlation with a positive SCA among 55 consecutive SOT recipients diagnosed with Cry at the University of Pittsburgh Medical Center. These included: heart (n=13), lung (n=4), liver (n=28), kidney (n=9) and small bowel (n=1) recipients. While there were no significant differences in the manifestations of Cry in heart and lung recipients, kidney recipients had disseminated disease as the most common presentation (P=0.02). In contrast, pneumonia (P=0.003) and meningitis (P=0.02) were more frequent than disseminated disease in liver recipients. Positive SCA was higher in patients with disseminated disease and meningitis than in patients with isolated pneumonia (P=0.0001). Significant differences in the manifestations of Cry were observed among types of SOT populations. A positive SCA may be predictive of dissemination and meningitis, but it may not be sensitive for pulmonary disease. PMID- 12759726 TI - The effects of a specific neutrophil elastase inhibitor (ONO-5046) in pulmonary ischemia-reperfusion injury. AB - Ischemia-reperfusion injury is a significant problem in lung transplantation. Polymorphonuclear elastase derived from neutrophils plays a major mechanistic role in this process. Hence, we have investigated the effects of ONO-5046, a neutrophil elastase inhibitor, on ischemia-reperfusion injury. Fifteen rabbits were divided into three groups: 2 h of single left-lung perfusion (control group, n=3); 2 h of ischemia followed by 2 h of reperfusion (ischemic group, n=6); and drip intravenous administration of ONO-5046 during the 2 h of ischemia and 2 h of reperfusion (ONO-5046 group, n=6). Hemodynamic parameters were determined and a histopathological examination of the lung was performed. In the ONO-5046 group, arterial oxygen pressure, cardiac output, and tissue blood perfusion were higher and pulmonary vascular resistance was lower than in the ischemic group. The ONO 5046 group also showed large decreases in neutrophil infiltration, pulmonary edema, and intra-alveolar hemorrhage. Treatment with ONO-5046 improves lung function in a rabbit-lung ischemia-reperfusion model. PMID- 12759727 TI - The effect of combined rapamycin/cyclosporine on the changes in pro-fibrotic gene expression that occur during the development of allograft vasculopathy in rats, compared with cyclosporine or rapamycin in isolation. AB - Chronic allograft dysfunction, the leading cause of solid-organ transplant failure, is characterised by histological evidence of extracellular matrix (ECM) accumulation (fibrosis). The aim of this study was to compare the effect of combined rapamycin and cyclosporine therapy on fibrosis-associated gene expression and ECM turnover during the development of allograft vasculopathy, compared with either agent alone. Lewis recipients of F344 rat thoracic-to abdominal aorta transplants were administered rapamycin, cyclosporine, combined rapamycin and cyclosporine or no treatment. F344-to-F344 isografts served as controls. Six grafts in each group were harvested at 16 weeks. Vascular remodelling and ECM accumulation (Sirius red) were measured by computerised histomorphometry of aortic sections. Messenger RNA was extracted from frozen tissue, and expression of fibrosis-associated genes was studied by means of semi quantitative reverse transcription polymerase chain reaction (RT-PCR). Rapamycin (0.5 mg/kg per day) or cyclosporine (5 mg/kg per day) inhibited intimal hyperplasia, medial ECM accumulation and expansive vascular remodelling (increasing vessel circumference) in rat aortic allografts. This was associated with attenuation of the graft inflammatory infiltrate and a reduction in intra graft gelatinase, collagen III and tissue inhibitor of metalloproteinase (TIMP)-1 mRNA levels. Combined rapamycin and cyclosporine inhibited intimal hyperplasia; however, there was a lesser effect on vascular remodelling and medial ECM accumulation. Combined-treatment aortic allografts were also seen to have a more severe inflammatory infiltrate and larger amounts of intra-graft matrix metalloproteinase (MMP)-9, transforming growth factor (TGF)-beta and TIMP-1 mRNA than those treated with monotherapy. Rapamycin and cyclosporine act synergistically to inhibit intimal hyperplasia but not the inflammatory infiltrate, allograft fibrosis or vessel remodelling. In the high-responder F344 to-Lewis rat model, effective immunosuppression is required to reduce graft fibrosis. PMID- 12759728 TI - Right-side approach to the left caudate lobe in the piggy-back technique of liver transplantation. A further step in maximizing preservation of the retrohepatic inferior vena cava. PMID- 12759729 TI - Characterisation of the phosphoenolpyruvate carboxylase gene family in sugarcane (Saccharum spp.). AB - Phosphoenolpyruvate carboxylases (PEPCs) are encoded by a small multigenic family. In order to characterise this gene family in sugarcane, seven DNA fragments displaying a high homology with grass PEPC genes were isolated using polymerase chain reaction-based cloning. A phylogenetic study revealed the existence of four main PEPC gene lineages in grasses and particularly in sugarcane. Moreover, this analysis suggests that grass C4 PEPC has likely derived from a root pre-existing isoform in an ancestral species. Using the Northern-dot blot method, we studied the expression of the four PEPC gene classes in sugarcane cv. R570. We confirmed that transcript accumulation of the C4 PEPC gene (ppc-C4) mainly occurs in the green leaves and is light-induced. We also showed that another member of this gene family (ppc-aR) is more highly transcribed in the roots. The constitutive expression for a previously characterised gene (ppc-aL2) was confirmed. Lastly, the transcript accumulation of the fourth PEPC gene class (ppc-aL1) was not revealed. Length polymorphism in non-coding regions for three PEPC gene lineages enabled us to develop sequence-tagged site PEPC markers in sugarcane. We analysed the segregation of PEPC fragments in self-pollinated progenies of cv. R570 and found co-segregating fragments for two PEPC gene lineages. This supports the hypothesis that diversification of the PEPC genes involved duplications, probably in tandem. PMID- 12759730 TI - Genetic basis of heterosis explored by simple sequence repeat markers in a random mated maize population. AB - The genetic basis of heterosis in crop plants has not been completely resolved. Our objective in this study was to determine the level of dominance for quantitative trait loci (QTLs) that underlie heterosis in maize (Zea mays L.). An F2 population of an elite maize single cross, LH200 x LH216, was random mated for three generations in an attempt to break up repulsion linkages that might lead to pseudo-overdominance. The population was analyzed with 160 simple-sequence repeat markers. Phenotypic data analyses indicated overdominance for grain yield and partial dominance for plant height, grain moisture and stalk lodging. A total of 28 QTLs were identified for grain yield, 16 for grain moisture, 8 for stalk lodging, and 11 for plant height. For grain yield, 24 QTLs (86%) showed overdominance. In contrast, most of the QTLs for plant height, grain moisture and stalk lodging showed partial to complete dominance. Little epistasis was detected among the QTLs for any of the traits. Our results can be interpreted in one of two ways, or a combination of both: (1) QTLs for grain yield in maize exhibit true overdominance, or (2) QTLs for grain yield show partial to complete dominance, but they are so tightly linked such that three generations of random mating failed to separate their individual effects. PMID- 12759731 TI - Identification of QTLs conferring resistance to downy mildews of maize in Asia. AB - Downy mildew is one of the most destructive diseases of maize in subtropical and tropical regions in Asia. As a prerequisite for improving downy mildew resistance in maize, we analyzed quantitative trait loci (QTLs) involved in resistance to the important downy mildew pathogens--Peronosclerospora sorghi (sorghum downy mildew) and P. heteropogoni (Rajasthan downy mildew) in India, P. maydis (Java downy mildew) in Indonesia, P. zeae in Thailand and P. philippinensis in the Philippines--using a recombinant inbred line population derived from a cross between Ki3 (downy mildew resistant) and CML139 (susceptible). Resistance was evaluated as percentage disease incidence in replicated field trials at five downy mildew 'hotspots' in the four countries. Heritability estimates of individual environments ranged from 0.58 to 0.75 with an across environment heritability of 0.50. Composite interval mapping was applied for QTL detection using a previously constructed restriction fragment length polymorphism linkage map. The investigation resulted in the identification of six genomic regions on chromosomes 1, 2, 6, 7 and 10 involved in the resistance to the downy mildews under study, explaining, in total, 26-57% of the phenotypic variance for disease response. Most QTL alleles conferring resistance to the downy mildews were from Ki3. All QTLs showed significant QTL x environment interactions, suggesting that the expression of the QTL may be environment-dependent. A strong QTL on chromosome 6 was stable across environments, significantly affecting disease resistance at the five locations in four Asian countries. Simple-sequence repeat markers tightly linked to this QTL were identified for potential use in marker assisted selection. PMID- 12759732 TI - Transgressive segregation of erucic acid content in Brassica carinata A. Braun. AB - Two Ethiopian mustard ( Brassica carinata A. Braun) lines with low (about 10%) and zero erucic acid (C22:1) have been obtained. The low C22:1 mutant line L-2890 was isolated after a chemical-mutagen treatment of C-101 seeds (about 40% C22:1). The zero C22:1 line L-25X-1 was obtained by interspecific crossing. Our objective was to determine the genetic control of low and zero C22:1 contents in these lines and the relationship between the loci controlling these traits. Reciprocal crosses between L-2890, L-25X-1 and high C22:1 lines, and between L-2890 and L 25X-1, were made. The F(1), F(2) and BC(1) F(1) generations were obtained. No maternal or cytoplasmic effects for C22:1 content were observed in any of the crosses. The analysis of the fatty acid composition in the segregating populations from the crosses of L-2890 with the high C22:1 lines C-101 and L-1630 indicated that the segregation patterns fitted a model of two alleles at two loci, M1 and M2, with partial (near complete) dominance for high concentration. The segregation patterns in the cross of the zero C22:1 line L-25X-1 with the high C22:1 line L-1630, were explained on the basis of two genes, E1 and E2, with additive gene action. The F(1) and segregating generations of the crosses L-2890 x L-25X-1 showed a strong transgressive segregation with C22:1 values of up to 50.0%, four-fold higher than those of L-2890. The analyses of the F(2), BC(1)F(1) and F(3) generations indicated that the combination of alleles at four loci, M(1) and M(2) in L-2890 and E(1) and E(2) in L-25X-1, controlled the transgressive segregation for C22:1. The proposed genotypes (C22:1 content) for each parent were as follows: L-2890 (10% C22:1) = m(1) m(1) m(2) m(2) E(1) E(1) E(2) E(2); L 25X-1 (0% C22:1) = M(1) M(1) M(2) M(2) e(1) e(1) e(2) e(2); and C-101 (45% C22:1) = M(1) M(1) M(2) M(2) E(1) E(1) E(2) E(2). PMID- 12759733 TI - [Sun protection during holidays]. AB - Ultraviolet radiation is causally involved in induction of skin cancer, premature skin aging and photodermatoses. The longing of our western society for a "healthy tanning" as well as the unbroken trend to spend the holidays in sunny regions lead to the fact that human skin is increasingly exposed to ultraviolet radiation and its detrimental effects. Because of the socio-political importance of the vacation period as the "most beautiful and most important time of the year", effective prevention of these unwanted UV effects has an enormous importance to the general population. In this article the most important methods for effective sun protection are critically discussed. PMID- 12759734 TI - [Cutaneous leishmaniasis]. AB - Leishmaniasis is a vector-borne disease caused by an obligate intracellular protozoa, Leishmania, which resides in macrophages. The parasite is transmitted by an infected female sandfly. The incidence of cutaneous leishmaniasis approaches 2 million new cases per year with 90% of the cases occurring in the "Old World", while the "New World" accounts for the rest. Infection may be restricted to the skin with development of characteristic ulcers, or may affect the mucous membranes in its mucocutaneous form. The clinical diagnosis is verified by the presence of amastigotes in slit-skin smears. Therapeutic modalities include systemic treatments such as the pentavalent antimony compound sodium stibogluconate, liposomal formulations of amphotericin B, oral ketoconazole or itraconazole, as well as topical paromomycin sulphate, local heat, freezing with liquid nitrogen, or photodynamic therapy. An effective vaccine is not available. PMID- 12759735 TI - [Onchocerciasis]. AB - Onchocerciasis is an infestation caused by the nematode, Onchocerca volvulus, and characterized by eye manifestations, skin lesions and troublesome itching. Although partially controlled by international mass treatment programs, onchocerciasis remains a major health hazard in endemic areas in Africa, Arabia, and the Americas. Onchocerciasis is spread by bites from infested blackflies which transmit larvae that subsequently develop into adult filariae. Skin findings are commonly non-specific, and include severe pruritus, acute and chronic dermatitis, vitiligo-like hypopigmentation and atrophy. Onchocercal ocular disease has a large spectrum of manifestations and may even lead to blindness. Diagnosis is usually made by direct visualization of the larvae emerging from superficial skin biopsies, "skin snips". In some cases, the microfilariae can also be directly observed with a slit lamp when they migrate into the anterior chamber of the eye. Ivermectin is highly microfilaricidal, and is the current drug of choice for both skin and ocular manifestations. PMID- 12759736 TI - [Traveler's thrombosis]. AB - Travel induced disorders have become more and more evident in a society of mass tourism. The link between travel in sitting position and risk of venous thromboembolic disease has been widely accepted. But the risk is not quantifiable because of a lack of epidemiological and pathophysiological data. It is likely to be small and mainly effect passengers with additional risk factors for venous thromboembolism. Therefore, prophylaxis of thromboembolic events during long distance journeys can only be considered good advice (with the exception of leg exercise) instead of an evidence-based recommendation. PMID- 12759737 TI - [Wound bed preparation of chronic wounds with ultrasound]. AB - The use of different low dose ultrasound systems is an innovative and effective alternative strategy of treatment in wound bed preparation of different chronic wounds. The handling of the ultrasonic systems is easy to learn, their use is safe and little additional equipment is required. Induction of the so called cavitation phenomenon seems to be a major effect which permits selective debridement and reduction of microorganism colonisation. While the suggestion is that granulation tissue is promoted and wound healing accelerated, there are few well documented evidence-based data. Additional randomized controlled trials are needed before the therapeutic efficacy of ultrasound in this clinical setting can be evaluated. PMID- 12759738 TI - [Somatization and alexithymia in male infertility. A replication study]. AB - BACKGROUND AND OBJECTIVE: In a previous study, we found a higher degree of alexithymia and somatization in infertile men (Hum Reprod 2001 Vol.16(3),587 592). This study was designed to confirm the findings of the former study PATIENTS/METHODS: 88 infertile men (48 idiopathic infertility, 40 somatic infertility) were consecutively enrolled in the study. The control group consisted of 44 healthy men. Alexithymia was measured by the 20-item-Toronto Alexithymia-Scale;somatization was measured by the Symptom Checklist-90-R. RESULTS: There were no differences between the subgroups of infertile men. The study group showed a significantly higher sum score in the TAS-20 (p<0.01) and a higher score on the scale somatization in the Symptom Checklist-90-R (p<0.05) compared to the control group. CONCLUSIONS: Our study replicated the findings of the previous investigation. Alexithymia and somatization are discussed as consequences of coping with male infertility. PMID- 12759739 TI - [Mycosis fungoides in childhood and adolescence with clonal T-cell receptor gamma gene rearrangement. Two cases]. AB - Mycosis fungoides (MF) is a cutaneous T-cell lymphoma (CTCL) characterized by its typical progress in three stages: the patch-, the plaque- and the tumour-stage. The incidence of mycosis fungoides rises with age and the average age at presentation is about 50. Children and adolescents are rarely affected and there are only few reports in the literature. We report a 12- and a 15-year-old boy showing refractory skin lesions not typical for mycosis fungoides. The histo- and immunohistological investigations and the detection of clonal T-cell receptor gamma gene rearrangements confirmed the diagnosis of early onset mycosis fungoides in both cases. PMID- 12759741 TI - [Multiple erythema]. PMID- 12759740 TI - [Malignant priapism as a sign of a recurrent prostate cancer. Differential diagnosis of induratio penis plastica]. AB - A 62-year-old patient was referred with the suspected diagnosis of Peyronie disease. The patient showed the clinical picture of a beginning malignant priapism with hematogenous metastases from an recurrent prostatic carcinoma. The penile metastases were misdiagnosed as Peyronie disease. Although both clinical pictures show some superficial similarity, Peyronie disease usually fulfills classical criteria which help to clearly distinguish it from neoplastic diseases. PMID- 12759742 TI - [The restored masterwork. The old man, his rhinophyma and the child]. AB - "The old man and the child" is a masterpiece by the Renaissance painter Domenico Ghirlandaio, which was restored a few years ago. We speculate as to the likely origin of the scratches on the old man's face and discuss the complicated restoration of the painting. PMID- 12759743 TI - [Excessive hyperchylomicronemia--a rare cause of acute retrosternal and epigastric pain in pregnancy]. AB - CASE REPORT: A 24-year-old woman in her 13th gestational was admitted to our department with acute retrosternal and epigastric pain. She had been transferred from the gynecologic department where she was treated for vaginal bleeding because of abortus imminens. A cardiac cause was excluded by ECG and echo. Clinical chemistry and abdominal ultrasound confirmed the diagnosis of acute pancreatitis. The woman was known in our outpatient department for hyperchylomicronemia and had already had an earlier episode of acute pancreatitis under oral contraception years ago. At current admission, triglycerides were 11,500 mg/dl. To reduce plasma triglycerides, selective lipid apheresis was performed. Apheresis was well tolerated, and the patient became free of pain within the first 30 min of treatment. Triglycerides decreased to 6,600 mg/dl at this session. Keeping to a low-fat diet (< 30 g fat per day), the patient remained healthy and completed pregnancy with the delivery of a healthy girl in her 39th week of pregnancy. CONCLUSION: Selective lipid apheresis is a safe and effective option in the treatment of hyperlipidemic pancreatitis, even in pregnant patients. PMID- 12759744 TI - Transcriptional analysis of the genetic elements involved in the lysogeny/lysis switch in the temperate lactococcal bacteriophage phiLC3, and identification of the Cro-like protein ORF76. AB - A transcriptional analysis of the lysogeny-related genes of the temperate bacteriophage Lactococcus lactis phiLC3 was performed using Northern blot hybridization during lysogeny and lytic infection by the phage. The lysogeny related gene cluster was found to contain four promoters (P(1), P(2), Pint and P(173)), while the P(87) promoter directed transcription of orf80 and the putative gene orf87, which are located between the integrase gene and the cell lysis genes. The start sites of the transcripts were determined by primer extension. The divergently oriented lysogenic P(1) and lytic P(2) promoters located in the genetic switch region are responsible for transcription of orf286 which encodes the phage repressor, and the genes orf63 - orf76 - orf236 - orf110 orf82 - orf57, respectively, while orf173 is transcribed from P(173). orf76 was identified as the gene encoding the Cro-like protein of phiLC3, and it was shown that ORF76 is able to bind specifically to the genetic switch region, albeit with lower affinity than does the phage repressor ORF286. ORF76 also competed with ORF286 for binding to this region. The functionality of P(1) and P(2), and their regulation by ORF286 and ORF76, was investigated using a reporter gene. In general, P(2) was a stronger promoter than P(1), but expression from both promoters, especially P(2), was regulated and modulated by flanking sequences and the presence of orf286 and orf76. ORF286 and ORF76 were both able to repress transcription from P(1) and P(2), while ORF286 was able to stimulate its own synthesis by tenfold. This work reveals the complex interplay between the regulatory elements that control the genetic switch between lysis and lysogeny in phiLC3 and other temperate phages of Lactococcus. PMID- 12759745 TI - Complexity and genetic diversity of Plasmodium falciparum infections in young children living in urban areas of Central and West Africa. AB - A site-based characterization of Plasmodium falciparum infections in children living in two malaria hyperendemic urban areas from West and Central Africa was undertaken. A total of 58 and 46 children with either asymptomatic infections or uncomplicated (symptomatic) malaria were recruited in Gabon and Benin, respectively. Parasite density, hematological factors, the genetic diversity of P. falciparum merozoite surface protein 2 (msp2) and the complexity of infections (mean number of P. falciparum genotypes per infected child) were used for this characterization. Gabonese children with uncomplicated malaria presented a higher mean axillary temperature (39.2 vs 38.6, P=0.004) and a higher geometric mean parasite density (30,538 vs 18,921, P<0.001) associated with a significantly lower hemoglobin level ( P<0.01). A higher degree of msp2 polymorphism and the complexity of P. falciparum infections were also observed in children from Gabon ( P<0.05). With a similar level of malaria transmission in both urban sites, these results suggest an impact of malaria control interventions on the dynamics of concurrent P. falciparum infections. PMID- 12759746 TI - Postlarval Protopolystoma spp. kidney infections in incompatible Xenopus spp. induce weak resistance to heterospecifics. AB - Protopolystoma xenopodis and Protopolystoma orientalis are polystomatid monogeneans respectively specific to the parapatric anurans Xenopus laevis and Xenopus muelleri. Parasite larval stages may invade the kidneys of foreign Xenopus spp. but die before migration to the definitive urinary bladder site. Laboratory experiments to assess the effect of a primary incompatible kidney infection on a secondary compatible infection found: (1) a small, significant decrease in the survivorship of P. xenopodis kidney stages (23-37 days p.i. at 25 degrees C) in X. laevis laevis previously challenged with P. orientalis; (2) a significant effect of prior P. orientalis challenge on P. xenopodis development and establishment in the urinary bladder of X. laevis 100 days p.i. (at 21 degrees C); (3) no effect of prior P. xenopodis challenge on adult P. orientalis establishment in X. muelleri (at 21 degrees C), but a significant negative influence on reproductive output (days 0-50 post-patency). Partial cross resistance to heterospecifics may therefore be induced by Protopolystoma spp. infections in the kidneys of an incompatible host, demonstrating that at least some elements of the host response are non-species specific. The effects observed were weak compared to the strong host resistance known to be generated by an established compatible primary infection with respect to conspecifics. This difference suggests that strong acquired resistance to Protopolystoma species is species-specific and/or induced only by older stages surviving in compatible hosts. PMID- 12759747 TI - Genetic polymorphisms of tobacco- and alcohol-related metabolizing enzymes and the risk of hepatocellular carcinoma. AB - The effect of genetic polymorphisms for glutathione S-transferase ( GST) M1, GSTT1, GSTP1-1( GSTP1), cytochrome P450 2E1 ( CYP2E1) and aldehyde dehydrogenase 2 ( ALDH2) on the risk of hepatocellular carcinoma (HCC) was observed in 78 Japanese patients with HCC and 138 non-cancer hospital controls. We found a positive association between cumulative amounts of alcohol consumption (>/=600,000 ml in a lifetime) and the risk of HCC (OR=4.52, 95% CI 2.39-8.55). However, cigarette smoking was not significantly related to the risk of HCC (OR=1.23, 95% CI 0.57-2.68). The allelic frequencies of GSTM1, GSTT1, GSTP1, CYP2E1and ALDH2of HCC patients were not significantly different from those of controls when odds ratios were only adjusted for age and gender except for any 2 alleles of ALDH2in drinkers (OR=2.53, 95% CI 1.21-5.31). However, the frequency of any C2 alleles of CYP2E1and any 2 alleles of ALDH2were significantly higher than those of controls (OR=5.77, 95% CI 1.24-27.39, OR=9.77, 95% CI 1.63-58.60) when covariates including viremia were selected by using stepwise logistic regression analysis. We conclude that habitual alcohol drinking is likely to lead to an increased risk of HCC, and any C2alleles of CYP2E1as well as any two alleles of ALDH2were also associated with an increased risk of HCC. PMID- 12759748 TI - Analysis of DLC-1 expression in human breast cancer. AB - The chromosome region 8p12-p22 shows frequent allelic loss in many neoplasms, including breast cancer (BC). The DLC-1 gene, located on 8p21-p22, might be a candidate tumor suppressor gene in this region. To evaluate the involvement of DLC-1 in breast carcinogenesis we studied DLC-1 mRNA expression in a panel of 14 primary human BC and the corresponding normal breast cells as well as 8 BC cell lines. Low levels or absence of DLC-1 mRNA were observed in 57% of primary BC and 62.5% of BC cell lines, respectively. We could not find any correlation between DLC-1 mRNA expression and deletions at the DLC-1 locus. Transfection of the gene into DLC-1 deficient T-47D cells raised the DLC-1 mRNA level and resulted in inhibition of cell growth and reduced colony-forming capacity. Our results indicate a role of DLC-1 in BC carcinogenesis. PMID- 12759749 TI - Stomatin immunoreactivity in ciliated cells of the human airway epithelium. AB - Stomatin is a widely distributed 32kD membrane protein of unknown function. In biochemical studies it is associated with cholesterol+sphingomyelin-rich 'rafts' in the cytomembrane. Genetic studies in C. elegans, supported by microscopic studies in mammalian tissue and co-expression studies in oocytes, suggest a functional link with the DEG/ENaC (degenerin/epithelial Na+ channel) superfamily of monovalent ion channels. Since ENaC channels play a prominent role in the physiology of the respiratory epithelium, we have studied the immunolocalization of stomatin in mature and developing human airway epithelium by means of Western blot analysis, immunocytochemistry, and immunoelectron microscopy. Stomatin immunoreactivity (stomatin-IR) was found in the ciliated cells of the conductive airway epithelium in a distinct distribution pattern with the strongest signal along the cilia. Immunogold labelling revealed immunogold particles at the basal bodies, along the cilia, and at the membrane of the microvilli. The presence of stomatin-IR paralleled the stages of ciliogenesis in airway development, and its appearance preceded the elongation of the axoneme and the cilial outgrowth. Due to its presence in the different cellular locations in the ciliated cell, we suggest that stomatin is involved in various cellular functions. From its ultrastructural position, stomatin could be a candidate for a membrane-associated mechanotransducer with a role in the control of ciliary motility. Stomatin as a raft protein might be a microtubule associated protein moving along the outer surface of the microtubules to its terminal site of action in the cilia. Stomatin IR in microvilli supports the hypothesis of a co-localization with beta- and gamma- ENaC and, in conclusion, their potential functional interaction to control the composition of periciliary mucus electrolytes. PMID- 12759750 TI - The reappraisal of gastrointestinal stromal tumors: from Stout to the KIT revolution. AB - For five decades gastrointestinal stromal tumors (GISTs) truly have represented one of the most confusing as well as neglected areas of both surgical pathology and clinical oncology. The recognition of the central role played by KIT expression in the development of the interstitial cell of Cajal and of the activating KIT mutations in the pathogenesis of GIST have been the keys for a more precise categorization of this long elusive clinicopathological entity. A Consensus Conference held at the National Institutes of Health in 2001 provided both an evidence-based definition and a practical scheme for the assessment of the risk of aggressive clinical behavior. This scheme is based on evaluation of the size and mitotic rate of the tumors, and its use is strongly advocated. On the basis of current data GISTs can be defined as a distinctive group of KIT expressing mesenchymal neoplasms of the gastrointestinal tract, showing differentiation towards the interstitial cell of Cajal, also known as the gastrointestinal pacemaker cells. Metastatic GISTs have been a virtually incurable disease until the elucidation of the role of KIT mutations. STI-571 (imatinib mesylate) is a molecule that inhibits the function of various receptors with tyrosine kinase activity, such as abl, the bcr-abl chimeric product, platelet-derived growth factor receptor, and KIT. Following its successful use in the treatment of chronic myeloid leukemia, STI-571 has also proved extremely effective in targeting metastatic GIST. Data regarding the duration of the response to this therapy are not yet available, and therefore any overenthusiasm should be avoided. Nonetheless, the GIST story remains paradigmatic of a totally innovative approach to cancer therapy which until now is the most elegant translation of cancer biology experimental knowledge into clinical practice. PMID- 12759751 TI - Altered expression of desmosomal components in high-grade squamous intraepithelial lesions of the cervix. AB - We have used immunohistochemistry to test the hypothesis that components of the desmosome are disrupted during neoplastic progression of squamous epithelial cells in the uterine cervix. Sections of normal cervix and squamous intraepithelial lesions (SILs) were immunostained for desmosomal proteins and glycoproteins, and results were assessed using a semi-quantitative grading system. No difference between normal cervix and low-grade SIL (LSIL) was found. A significant reduction in expression of desmogleins was seen between high-grade SIL (HSIL) and LSIL (P<0.01) and normal cervix (P<0.001). Desmocollin expression was not reduced significantly, although scores showed significantly greater variation in HSIL compared with LSIL (P<0.05) and normal cervix (P<0.05). There was no significant difference in desmoplakin expression among the three groups. The results suggest that there may be sequential disruption of desmosomal function during neoplastic progression of cervical squamous intraepithelial cells, with downregulation of desmogleins during the progression from LSIL to HSIL and loss of desmocollin expression occurring in some cases of established HSIL. PMID- 12759752 TI - A four-nucleotide base-pair deletion in the coding region of the Bowman-Birk protease inhibitor gene prevents its accumulation in the seeds of Glycine microphylla PI440956. AB - The Bowman-Birk protease inhibitor (BBI), an abundant soybean [ Glycine max (L.) Merr.] seed protein, is a major antinutritional factor. Nulls for the major soybean BBI have been reported in several of the wild perennial Glycine species including G. microphylla (Benth.) Tind PI440956. This perennial Glycine species does not accumulate the major BBI and the molecular basis for the absence of the major BBI in this plant introduction (PI) line is not known. We have cloned the BBI gene from G. microphylla PI440956, G. microphylla PI505188, and G. max cv. Jefferson and determined its nucleotide sequences. Analysis of the G. microphyllla PI505188 and G. max cv. Jefferson nucleotide sequences revealed a complete open-reading frame encoding the BBI. In contrast, the BBI coding region of G. microphylla PI440956 contained a frameshift mutation that resulted in the introduction of a stop codon at the amino terminal region of the protein. Reverse transcription-polymerase chain reaction analysis revealed that the BBI gene was expressed in developing seeds of G. microphylla PI505188 and G. max cv. Jefferson, but not in developing seeds of G. microphylla PI440956. In contrast, a BBI-related isoinhibitor gene was expressed at similar levels in all three Glycine species. Our results suggest that the frameshift mutation in the BBI coding region is responsible for the absence of BBI in the seeds of G. microphylla PI440956. PMID- 12759753 TI - Basolateral localization of flounder Na+-dicarboxylate cotransporter (fNaDC-3) in the kidney of Pleuronectes americanus. AB - The purpose of this study was to provide functional and immunocytochemical evidence for the location of the winter flounder ( Pleuronectes americanus) sodium-dicarboxylate cotransporter-3 (fNaDC-3) in the basolateral membrane of proximal tubule cells. fNaDC-3 was expressed in Xenopus laevis oocytes. Lowering the external pH from 7.5 to 6.5 or 5.5 modestly decreased the uptake of [(14)C]succinate into fNaDC-3 expressing oocytes, but markedly increased the uptake of [(14)C]citrate. As measured by the two-electrode voltage-clamp technique, the citrate concentration eliciting half-maximal current, K(0.5), decreased from 490 microM at pH 7.5 to 32 microM at pH 6.0. The maximal inwards current, Delta I(max), increased from -27 to -72 nA, when bath pH was changed from 7.5 to 6.0. These data suggest that fNaDC-3 translocates preferably divalent citrate. cis-Aconitate, a tricarboxylate that interacts exclusively with basolateral sodium-dicarboxylate cotransport in the rat kidney, was translocated by fNaDC-3 with a K(0.5) of 300 microM. Antibodies raised against an NaDC-3 specific peptide reacted with the basal cell side of flounder renal proximal tubule segment II (PII). No other structures were stained, indicating that fNaDC 3 is located exclusively in the basolateral membrane of PII cells. We assume that fNaDC-3 provides PII cells with Krebs cycle intermediates as fuels and with alpha ketoglutarate to drive organic anion secretion. PMID- 12759754 TI - The SLC20 family of proteins: dual functions as sodium-phosphate cotransporters and viral receptors. AB - The SLC20 family transport proteins were originally identified as retroviral receptors (called Glvr-1 and Ram-1). Since then, they have been shown to function as sodium-phosphate (Na/P(i)) cotransporters, and have subsequently been classified as type III Na/P(i) cotransporters (now called Pit-1 and Pit-2). The Pit cotransporters share approximately 60% sequence homology, they have a high affinity for P(i), they are electrogenic with a coupling stoichiometry of >1 Na(+) per P(i) ion cotransported, and are inhibited by alkaline pH and phosphonoformic acid (PFA). Pit-1 and Pit-2 expression and/or activity has also been shown to be regulated by P(i) deprivation in some, but not all cells and tissues examined. The Pit-1 and Pit-2 cotransporters are widely expressed, but cell-type specific expression has only been investigated in bone, kidney and intestine. Both proteins are likely expressed on the basolateral membranes of polarized epithelial cells, where they are likely involved in cellular P(i) homeostasis. The Pit-1 and Pit-2 gene promoters have been cloned and characterized. While the exact roles of the Pit cotransporters in different cell types has not been definitively determined, they may be involved in important physiological pathways in bone, aortic smooth muscle cells, parathyroid glands, kidney and intestine. PMID- 12759755 TI - The SLC26 gene family of multifunctional anion exchangers. AB - The ten-member SLC26 gene family encodes anion exchangers capable of transporting a wide variety of monovalent and divalent anions. The physiological role(s) of individual paralogs is evidently due to variation in both anion specificity and expression pattern. Three members of the gene family are involved in genetic disease; SLC26A2 in chondrodysplasias, SLC26A3 in chloride-losing diarrhea, and SLC26A4 in Pendred syndrome and hereditary deafness (DFNB4). The analysis of Slc26a4-null mice has significantly enhanced the understanding of the roles of this gene in both health and disease. Targeted deletion of Slc26a5 has in turn revealed that this paralog is essential for electromotor activity of cochlear outer hair cells and thus for cochlear amplification. Anions transported by the SLC26 family, with variable specificity, include the chloride, sulfate, bicarbonate, formate, oxalate and hydroxyl ions. The functional versatility of SLC26A6 identifies it as the primary candidate for the apical Cl(-) formate/oxalate and Cl(-)-base exchanger of brush border membranes in the renal proximal tubule, with a central role in the reabsorption of Na(+)-Cl(-) from the glomerular ultrafiltrate. At least three of the SLC26 exchangers mediate electrogenic Cl(-)-HCO(3)(-) and Cl(-)-OH(-) exchange; the stoichiometry of Cl(-) HCO(3)(-) exchange appears to differ between SLC26 paralogs, such that SLC26A3 transports >/=2 Cl(-) ions per HCO(3)(-) ion, whereas SLC26A6 transports >/=2 HCO(3)(-) ions per Cl(-) ion. SLC26 Cl(-)-HCO(3)(-) and Cl(-)-OH(-) exchange is activated by the cystic fibrosis transmembrane regulator (CFTR), implicating defective regulation of these exchangers in the reduced HCO(3)(-) transport seen in cystic fibrosis and related disorders; CFTR-independent activation of these exchangers is thus an important and novel goal for the future therapy of cystic fibrosis. PMID- 12759756 TI - Molecular physiology and pathology of the nucleotide sugar transporter family (SLC35). AB - The solute carrier family SLC35 consists of at least 17 molecular species in humans. The family members so far characterized encode nucleotide sugar transporters localizing at the Golgi apparatus and/or the endoplasmic reticulum (ER). These transporters transport nucleotide sugars pooled in the cytosol into the lumen of these organelles, where most glycoconjugate synthesis occurs. Pathological analyses and developmental studies of small, multicellular organisms deficient in nucleotide sugar transporters have shown these transporters to be involved in tumour metastasis, cellular immunity, organogenesis and morphogenesis. Leukocyte adhesion deficiency type II (LAD II) or the congenital disorder of glycosylation type IIc (CDG IIc) are the sole human congenital disorders known to date that are caused by a defect of GDP-fucose transport. Along with LAD II, the possible involvement of nucleotide sugar transporters in disorders of connective tissues and muscles is also discussed. PMID- 12759758 TI - Cardiorespiratory and hemodynamic responses during repetitive incremental lifting and lowering in healthy males and females. AB - The purposes of this study were twofold. First, to evaluate the cardiorespiratory and muscle oxygenation (OXY)/blood volume (BV) responses during repetitive incremental lifting and lowering (RILL) in healthy males and females. Second, to develop a predictive equation for predicting peak aerobic power (VO(2peak)) during RILL from the cardiorespiratory, OXY/BV and body composition variables. Fourteen males and 18 females [mean (SD) for age, height and body mass were: 29.6 (8.2) years; 1.75 (0.07) m; 78.9 (10.4) kg and 23.9 (2.1) years; 1.63 (0.06) m; 62.3 (6.3) kg, respectively] completed a RILL from floor to table height at 10 lifts/min to voluntary fatigue. Cardiorespiratory responses were measured using open circuit spirometry and hemodynamic trends were monitored bilaterally at the third lumbar vertebra via near infrared spectroscopy. Significant sex differences ( p<0.05) were observed for the peak values of oxygen uptake (VO(2peak)), ventilation rate (V(E)), oxygen pulse, BV-max and BV-delta. Erector spinae OXY decreased systematically until VO(2peak )was attained, while BV decreased until approximately 50% of VO(2peak) and then leveled off. Stepwise regression analysis indicated that approximately 75% of the variance in VO(2peak )was predicted from cardiorespiratory, hemodynamic and body composition variables, with the most important predictors for absolute and relative VO(2peak )being V(E) ( r=0.75) and fat mass ( r=-0.63) respectively. Inclusion of left side OXY/BV responses increased the predictability of the common variance in VO(2peak )from 40% to 74%, implying that muscle hemodynamics play an important role in determining VO(2peak )during RILL. PMID- 12759757 TI - Parallel down-regulation of chloride channel CLC-K1 and barttin mRNA in the thin ascending limb of the rat nephron by furosemide. AB - In the past few years the pivotal role of kidney Cl(-)channels (ClC-K) channels in maintaining salt and water homeostasis in the kidney has been established. The aim of the present study was to investigate the influence of the loop diuretic furosemide on the gene expression of the kidney chloride channel ClC-K1 and its recently described functional subunit barttin. Male Sprague Dawley rats received the loop diuretic furosemide (12 mg/kg/day) for 6 days. Rats had free access to 0.9% NaCl, 0.1%KCl solution to prevent volume depletion. Localisation and regulation of ClC-K1 and barttin mRNA was analysed by RNase protection and in situ hybridisation. Nephron-specific regulation was investigated by microdissection and real-time PCR quantification. In furosemide-treated rats ClC K1 mRNA decreased to half in the inner medulla. In the renal cortex and outer medulla ClC-K1 mRNA levels were weak and did not change. Under furosemide treatment barttin mRNA was regulated in parallel with ClC-K1 mRNA. A significant mRNA decrease occurred after furosemide treatment in inner medulla (0.50 fold), whereas cortical and outer medulla levels remained unaffected. (35)S in situ hybridisation confirmed the regulation and distribution seen in the RNase protection assay experiments. Microdissection of the inner medullary collecting duct and thin limb of Henle's loop followed by real-time PCR revealed that CLC-K1 and barttin mRNA regulation in inner medulla was limited to the thin limb; mRNA levels in collecting ducts were not affected by furosemide treatment. Our findings imply that during furosemide treatment selective down-regulation of ClC K1 and barttin mRNAs in thin limb plays a role in maintaining salt and water homeostasis. PMID- 12759759 TI - Oxygen uptake kinetics during severe intensity running and cycling. AB - The purpose of this study was to investigate the effect of exercise mode on the characteristics of the oxygen uptake (VO(2)) ()response to exercise within the severe intensity domain. Twelve participants each performed a treadmill running test and a cycle ergometer test to fatigue at intensities selected to elicit a mode-specific VO(2)max and to cause fatigue in ~5 min. The tests were at 234 (30) m.min(-1) and 251 (59) W, and times to fatigue were 297 (15) s and 298 (14) s, respectively. The overall rapidity of the VO(2)response was influenced by exercise mode [VO(2)max was achieved after 115 (20) s in running versus 207 (36) s in cycling; p<0.01]. VO(2) responses were fit to a three-phase exponential model. The time constant of the primary phase was faster in treadmill tests than in cycle ergometer tests [14 (6) s versus 25 (4) s; p<0.01], and the amplitude of the primary phase was greater in running than in cycling when it was expressed in absolute terms [2327 (393) ml.min(-1) versus 2036 (301) ml.min(-1); p=0.02] but not when it was expressed as a percentage of the total increase in VO(2) [86 (6)% versus 82 (6)%; p=0.09]. When quantified as the difference between the end exercise VO(2) and the VO(2) at 2 min, the amplitude of the slow component was ~40% smaller in running [177 (92) ml.min(-1) versus 299 (153) ml min(-1); p=0.03]. It is concluded that exercise modality affects the characteristics of the VO(2) response at equivalent intensities in the severe domain. PMID- 12759760 TI - Low frequency of the "plateau phenomenon" during maximal exercise in elite British athletes. AB - A plateau in oxygen consumption (VO(2)) has long been considered the criterion for maximal effort during an incremental exercise test. But, surprisingly, the termination of a maximum exercise test often occurs in the absence of a VO(2) plateau. To explain this inconsistency, some have proposed that an oxygen limitation in skeletal muscle occurs only in elite athletes. To evaluate this hypothesis, we determined the frequency with which the "plateau phenomenon" developed in a group of elite male and female athletes. Fifty subjects performed a continuous incremental treadmill test to measure maximal oxygen consumption (VO(2max)). Treadmill velocity increased by 0.31 m s(-1) until the respiratory exchange ratio ( R) reached 1.00. Thereafter the treadmill gradient increased by 1% each minute until exhaustion. The VO(2max) was the highest VO(2) sustained for 60 s. Three criteria were used to determine maximal efforts: (1) a plateau in the VO(2), defined as an increase of less than 1.5 ml kg(-1) min(-1); (2) a final R of 1.1 or above; (3) a final heart rate (HR) above 95% of the age-related maximum. Mean VO(2max) exceeded 65 ml kg(-1) min(-1) in both groups. The criteria for R and HR were satisfied by 72% of males and 56% females, and 55% of males and 69% of females, respectively. In contrast a VO(2) plateau was identified in only 39% of males and 25% of females. These findings refute the twin arguments: (1) that the absence of a "plateau phenomenon" results from an inadequate motivational effort in poorly trained athletes and (2) that the "plateau phenomenon" and a consequent skeletal muscle anaerobiosis occur only in athletes with the highest VO(2max) values. PMID- 12759762 TI - Adenosquamous carcinoma of the tongue in a 22-year-old female: report of a case with immunohistochemistry. AB - Adenosquamous carcinoma is a very rare tumor that is characterized pathologically by the simultaneous presence of distinct areas of adenocarcinoma and squamous cell carcinoma. Only 15 cases in the oral cavity are reported in the literature, most occurring in middle-aged and elderly males. Pain is a commonly reported presenting symptom and may be related to the frequent presence of perineural invasion. Most cases have been treated surgically. However, early recurrence and death due to disease are common. We report an exceptional case occurring in a 22 year-old female. The tumor was associated with dysplasia of both minor salivary gland ductal epithelium and surface mucosal epithelium. Perineural invasion was also present. The patient was treated with combined surgery and radiotherapy and remained well for 9 months before developing locoregional recurrence and ultimately succumbing to the disease. PMID- 12759761 TI - Blood glucose threshold and the metabolic responses to incremental exercise tests with and without prior lactic acidosis induction. AB - This study compared the metabolic-ventilatory responses and the glycemic threshold identified during lactate minimum (LM) and individual anaerobic threshold (IAT) tests. In addition, the ability to determine the anaerobic power, aerobic-anaerobic transition (Trans) (e.g. ventilatory threshold; VT) and the maximal oxygen consumption (VO(2max)) all within a single incremental treadmill test (IT) was investigated. Fifteen physically fit men [25.9 (5.5) years; 77.4 (6.5) kg] performed the following: test 1, IT for IAT; and test 2, LM: 30-s Wingate test followed by 8 min rest and then an IT that was the same as test 1. Blood lactate concentration [lac], glucose concentration [gluc], pH, PO(2), PCO(2), base excess (BE) and ventilatory variables were measured. At the beginning of the IT for LM, the ventilation, PO(2) and VO(2) were higher and the pH, BE and PCO(2) were lower in relation to IAT ( P<0.05), while no differences were observed after reaching LM intensity during IT. Moreover, the Trans could be identified by [lac] (IAT, LM), minute ventilation [V(E;) VT identified during IAT protocol (VT-IAT) and VT identified during LM protocol (VT-LM)], and [gluc] (IGT, GM) during the IT for IAT and LM. The velocities (kilometers per hour) corresponding to IAT (12.6+/-1.6), VT-IAT (12.5+/-1.7), IGT (12.6+/-1.6), LM (12.5+/-1.5), VT-LM (12.3+/-1.5), and GM (12.6+/-1.9) were not different from each other and the LM and IAT protocols resulted in the similar VO(2max). We concluded that: (1) after reaching the LM the metabolic responses during IT are similar to IAT; (2) performing a Wingate test prior to an IT does not interfere with the Trans and VO(2max) attainment; (3) and the IGT and GM can predict the Trans. PMID- 12759763 TI - Hemangiopericytoma: a rare head and neck tumor. AB - Among the rare malignant tumors of the paranasal sinuses and the middle ear, the hemangiopericytoma represents a very rare tumor entity. Reviewing four cases treated in our ENT department, we present here our experiences with this tumor. Inpatient and outpatient medical records from 1976 to 2001 were evaluated retrospectively. For the whole period we found four patients, three women and one man, who were treated with the diagnosis of hemangiopericytoma. Three tumors were localized in the paranasal sinuses (two in the maxillary sinus, one in the frontal sinus) and one in the middle ear. While the two tumors of the maxillary sinus and the tumor of the middle ear were treated only by surgery, the patient with the tumor of the frontal sinus also received postoperative irradiation because of an endocranial infiltration. This patient was the only one who developed an endocranial recurrence 14 years later on the opposite site, which again was treated with a combination of surgery and radiotherapy. The hemangiopericytoma as a very rare tumor in the paranasal sinuses and the middle ear should be treated with surgery. In our view, an adjuvant radiotherapy is indicated only in selected individual cases. PMID- 12759764 TI - Increased expression of ICAM-1 and VCAM-1 in the lung of nitrofen-induced congenital diaphragmatic hernia in rats. AB - Recently, increased expression of inflammatory cytokine, tumor necrosis factor (TNF)-alpha, has been reported in both humans and animal models with CDH and the decreased TNF-alpha expression in CDH lung after antenatal dexamethasone (Dex) treatment. Intercellular adhesion molecule (ICAM)-1 and vascular cell adhesion molecule (VCAM)-1 are induced by several inflammatory cytokines such as TNF alpha. The aim of this study was to investigate pulmonary ICAM-1 and VCAM-1 expression in CDH lung in rats and to determine the effect of antenatal glucocorticoid. CDH model was induced in pregnant rats following administration of nitrofen on day 9.5 of gestation. In control animals, the same dose of olive oil was given without nitrofen. Dex (0.25 mg/kg) was given on day 18.5 and 19.5 of gestation. RT-PCR was performed to evaluate the relative amount of ICAM-1 and VCAM-1 mRNA expression. Fluorescein immunohistochemistry using anti-ICAM-1 and anti-VCAM-1 antibody was performed using light and confocal microscopy. ICAM-1 and VCAM-1 mRNA expression and ICAM-1 and VCAM-1 immunoreactivity were markedly increased in CDH lung compared to controls. Dex downregulated the expression of both adhesion molecules in the hypoplastic lung. Increased ICAM-1 and VCAM-1 mRNA expression in hypoplastic lungs would suggest that the increased local synthesis of pulmonary adhesion molecules may induce respiratory distress in CDH. Decreased expression of adhesion molecules in CDH lungs after Dex treatment suggests that antenatal glucocorticoids therapy may improve pulmonary immaturity and associated respiratory distress in nitrofen-induced CDH lung. PMID- 12759765 TI - ICAM-1 expression is upregulated in reflux nephropathy. AB - Reflux nephropathy (RN) is a major cause of end-stage renal failure in children and young adults. Intercellular adhesion molecule-1 (ICAM-1), a cell surface glycoprotein, has a role in the regulation of interaction among immune cells. It has been demonstrated that increased levels of tubular ICAM-1 correlate with an extent of tubular damage in diabetic nephropathy. We hypothesized that ICAM-1 local synthesis is altered in reflux nephropathy and therefore designated this study to investigate ICAM-1 expression in RN. The kidney specimens from six patients with severe reflux nephropathy secondary to primary vesicoureteral reflux were obtained at the time of nephrectomy. Control materials included normal kidney specimens obtained from three adult patients during partial nephrectomy for an incidentaloma. Fluorescent immunohistochemistry was carried out using monoclonal antibodies to ICAM-1 utilizing confocal laser scanning microscopy. Reverse transcriptase polymerase chain reaction (RT-PCR) was performed to evaluate the relative amount of ICAM-1. In the control kidneys, there was lack of ICAM-1 immunoreactivity in the interstitium and proximal tubules and moderate immunoreactivity in the glomerulus. In the refluxing kidney there was strong ICAM-1 immunoreactivity in the glomerulus, interstitium and proximal tubules. The RT-PCR showed strong ICAM-1 mRNA expression in the refluxing kidneys and absent or weak ICAM-1 expression in the controls. Our findings of increased expression of ICAM-1 in the severe reflux nephropathy kidney suggests that ICAM-1 may play a role in the pathogenesis of renal parenchymal damage associated with RN. PMID- 12759766 TI - Effects of seasonal changes in food quality and food intake on the transport of sodium and butyrate across ruminal epithelium of reindeer. AB - Transport of 22Na and 14C-butyrate across the ruminal epithelium of captive reindeer fed a concentrate diet in summer (n=5) and in winter (n=5) and from free ranging reindeer taken from summer (n=3) and winter pasture (n=5) was measured in vitro in Ussing chambers. Significant amounts of both Na+ and butyrate were transported across the isolated epithelium without any external driving force. The ruminal transport of Na+ and butyrate were interacting, as evidenced by both the observed amiloride-induced reduction of net butyrate-transport and by the positive correlation between net transport of butyrate and Na+. Amiloride also reduced the net transport of Na+ without significantly affecting the short circuit current, indicating the presence of an apical Na+/H+ exchanger in the ruminal epithelium of reindeer. The captive reindeer increased the dry matter intake of a constant quality concentrate from winter to summer, but this neither affected their ruminal transport capacity nor their ruminal surface enlargement factor (SEF). Free-ranging reindeer increased their ruminal transport capacity for Na+ and butyrate from summer to winter but simultaneously reduced their ruminal SEF. The present data indicate that this food-induced increase in transport capacity was attributed to changes in the nutrient composition of the diet. PMID- 12759767 TI - Energy reserve utilization in northern elephant seal (Mirounga angustirostris) pups during the postweaning fast: size does matter. AB - During fasting most mammals preferentially utilize lipid reserves for energy while sparing protein reserves. This presents a potential problem for marine mammals that also depend on lipids as a major component of blubber, the primary thermoregulatory structure. Because of this dual function for lipid, rates of lipid and protein utilization should be closely regulated during the postweaning fast in northern elephant seals (Mirounga angustirostris). To quantify energy expenditure during the fast, we measured body mass and composition of 60 pups at 2.3+/-0.2 days and 55.9+/-0.3 days postweaning in 1999 and in 2000. Body condition differed significantly between years. At weaning, body mass (125.9+/ 3.8 kg) and percentage lipid content (39.3+/-0.6% of body mass) in 2000 were significantly greater than body mass (115.2+/-3.1 kg) and percentage lipid content (35.8+/-0.6%) in 1999. In general, percentage lipid content increased with body mass, and fatter pups utilized lipid at relatively higher rates during the fast. Lipid fueled 85-95% and 88-98% of energy expended by pups in 1999 and 2000, respectively. Postweaning fast duration (32-78 days) was positively correlated with body mass and hence lipid content at weaning. This suggests that body composition at weaning influences lipid utilization patterns and ultimately the duration of the postweaning fast in northern elephant seal pups. PMID- 12759768 TI - Changes in glucose, glycogen, thyroid activity and hypothalamic catecholamines in tench by starvation and refeeding. AB - The effects of short-term food deprivation (7 days) and refeeding (2 days) on different biochemical and neuroendocrine parameters were studied in tench. A 7 days fast resulted in a significant reduction of plasma glucose and glycogen hepatic content, supporting the key role of liver glycogen as energy depot for being consumed during fasting. The rapid recovery of normal values of blood glucose and glycogen stores by refeeding indicates a rapid replenishment of liver glycogen stores. The short-term starvation decreased circulating thyroid hormones (both T3 and T4) and T4 release from thyroid, supporting an interaction between nutritional state and thyroid function in tench. All these metabolic and hormonal changes were partial or totally reversed under refeeding conditions. An increase in hypothalamic content of norepinephrine and dopamine was found in fasted fish. This result might be a consequence of stress induced by starvation. PMID- 12759769 TI - To swim or not to swim: regional effects of serotonin, octopamine and amine mixtures in the medicinal leech. AB - Focally treating the head brain of the medicinal leech Hirudo medicinalis with various biogenic amines affected the initiation, termination and maintenance of fictive swimming (i.e., the neural correlate of swimming). Application of serotonin to saline surrounding only the head brain inhibited fictive swimming, whereas removing serotonin induced swimming. This contrasts sharply with previous observations that serotonin applied to the nerve cord induces swimming. Although application of octopamine to the brain activated swimming, a mixture of octopamine and serotonin inhibited swimming. Subsequent removal of this mixture from the brain activated robust swimming and was more potent for activating swimming than either the removal of serotonin or the application of octopamine. Swim episodes induced by brain-specific manipulations of octopamine had more swim bursts per episode than those induced by serotonin. These brain-specific effects of the amines on fictive swimming are probably due to the modulation of higher order circuits that control locomotion in the leech. We observed that serotonin or a mixture of serotonin and octopamine hyperpolarized an identified descending brain interneuron known as Tr2. Removal of the mixture caused Tr2 to exhibit membrane potential depolarizations that correlated in time with the expression of swim episodes. PMID- 12759770 TI - Diffusion-weighted imaging of bone marrow: current status. AB - Diffusion-weighted imaging allows for measurement of tissue microstructure and reflects the random motion of water protons. It provides a new method to study bone marrow and bone marrow alterations on the basis of altered water-proton mobility in various diseases. Different diffusion-weighted methods have proved to be capable of differentiating between benign edema and tumorous involvement of bone marrow. It is especially useful for the distinction of acute benign osteoporotic and malignant vertebral compression fractures. Diagnosis is based on the contrast to normal bone marrow. Hypo- or isointensity reflects acute benign collapse, whereas hyperintensity is indicative of the tumorous nature of a fracture. Apparent diffusion coefficients (ADC) are significantly lower in metastatic disease than in bone marrow edema. Furthermore, bone marrow cellularity can be estimated by ADC measurements. Diffusion-weighted imaging might be helpful for monitoring response to therapy in metastatic disease. PMID- 12759772 TI - The probability of malignancy in small pulmonary nodules coexisting with potentially operable lung cancer detected by CT. AB - The aim of this study was to assess the probability of malignancy in one or two small nodules 1 cm or less coexisting with potentially operable lung cancer (coexisting small nodules). The preoperative helical CT scans of 223 patients with lung cancer were retrospectively reviewed. The probability of malignancy of coexisting small nodules was evaluated based on nodule size, location, and clinical stage of the primary lung cancers. Seventy-one coexisting small nodules were found on conventional CT in 58 (26%) of 223 patients, and 14 (6%) patients had malignant nodules. Eighteen (25%) of such nodules were malignant. The probability of malignancy was not significantly different between two groups of nodules larger and smaller than 0.5 cm ( p=0.1). The probability of malignancy of such nodules within primary tumor lobe was significantly higher than that in the other lobes ( p<0.01). Metastatic nodules were significantly fewer in clinical stage-IA patients than in the patients with the other stage ( p<0.01); however, four (57%) of seven synchronous lung cancers were located in the non-primary tumor lobes in the clinical stage-I patients. Malignant coexisting small nodules are not infrequent, and such nodules in the non-primary tumor lobes should be carefully diagnosed. PMID- 12759771 TI - Ultra-low-dose coronary artery calcium screening using multislice CT with retrospective ECG gating. AB - The aim of this study was to reduce radiation exposure in multislice CT (MSCT) coronary artery calcium screening using different tube settings, and to determinate its impact on the detection and quantification of coronary artery calcification. Forty-eight patients underwent routine MSCT coronary artery calcium scoring (Somatom VolumeZoom, Siemens, Forchheim, Germany) with retrospective ECG-gated data acquisition. Scanning was performed with a 4 x 2.5 mm collimation. In each patient data acquisition was performed twice using tube settings of 120 kVp with 133 mAs (protocol 1) and of 80 kVp with 300 mAs (protocol 2). Together with the 80-kVp protocol additional online ECG-related tube current modulation (ECG pulsing) was used. Three-millimeter overlapping slices (increment 1.5 mm) were calculated for each data set. Semi-automated calcium quantification was performed calculating absolute Ca-hydroxylapatite mass. In addition to patient examinations, the radiation exposure for both protocols was evaluated using computed tomography dose index (CTDI) phantom measurements. Protocol 2 showed a significantly lower patient radiation exposure than protocol 1 (0.72 vs 2.04 mSv; p<0.0001). The CTDI phantom measurements revealed a 65% reduction of radiation dose. Calcium scoring results of both protocols showed a high correlation ( r=0.99; p<0.0001) for absolute Ca Hydroxylapatite mass measurements. Using 80-kVp protocols patient radiation exposure can be significantly reduced in MSCT coronary artery calcium screening without affecting the detection and quantification of coronary artery calcification; therefore, this technique should be used with retrospective ECG gated cardiac CT examinations in patients with regular sinus rhythm. PMID- 12759773 TI - Radiologic investigation after laparoscopic inguinal hernia repair. AB - Laparoscopic instead of open surgical repair of inguinal hernias is becoming more frequent. Radiologists may expect different postoperative findings depending on the technique used. We studied how radiology had been used postoperatively and what findings were encountered after laparoscopic herniorraphy. Postoperative radiologic examinations related to hernia repair of all consecutive patients that had had laparoscopic herniorraphy in Malmo University hospital between 1992 and 1998 were retrospectively evaluated. A total of 538 groins were included, 3.9% (n=21) of these were postoperatively examined with ultrasound (n=10), herniography (n=7), plain abdominal films (n=2), CT (n=1), or fistulography (n=1). Significant findings were found in five groins, namely, one sinus tract, two hematomas, one small bowel obstruction, and one recurrence of hernia. Four insignificant seromas were found. The characteristics of the findings and pitfalls are described. Symptoms resulting in radiologic examination are rare after laparoscopic herniorraphy. The radiologist must be familiar with the spectrum of such findings. PMID- 12759776 TI - Extent of parietal peritonectomy does not change intraperitoneal chemotherapy pharmacokinetics. AB - PURPOSE: To measure the clearance intraperitoneal mitomycin C and doxorubicin in patients having peritonectomy and analyze the impact of the extent of peritoneal resection on pharmacokinetics. METHODS: A group of 15 patients with peritoneal carcinomatosis were submitted to cytoreductive surgery and heated intraperitoneal chemotherapy. Ten patients received mitomycin C and five, doxorubicin. Six patients underwent total parietal peritonectomy and nine had less-extensive peritonectomy. Pharmacokinetics were determined by sampling peritoneal fluid and blood. Drug concentrations over time, area under the curve ratios and the amount of drug recovered from the peritoneal cavity were calculated and compared between the groups. RESULTS: The concentrations of mitomycin C over time in the peritoneal fluid and plasma were similar in five patients with total parietal peritonectomy as compared to five patients with less-extensive peritonectomy ( P=0.5350 and 0.6991; Mann-Whitney test). Mitomycin C area under the curve ratio in total peritonectomy patients was 20.5 and 25.7 in patients with less-extensive peritonectomy. The difference in total amount of drug recovered from the peritoneal cavity was not significant (30.6+/-6.188% versus 22.6+/-3.84%, P=0.095). In the studies with doxorubicin, one patient underwent total parietal peritonectomy with similar pharmacokinetics to four patients submitted to partial peritonectomy. CONCLUSIONS: The extent of parietal peritoneal resection did not affect the pharmacokinetics of intraoperative intraperitoneal chemotherapy. The pharmacological barrier between the abdominopelvic cavity and plasma is not directly related to an intact peritoneum. PMID- 12759775 TI - Analysis of docetaxel pharmacokinetics in humans with the inclusion of later sampling time-points afforded by the use of a sensitive tandem LCMS assay. AB - PURPOSE: Docetaxel is a semisynthetic taxane derived from the needles of the European yew ( Taxus baccata) and it is an important chemotherapeutic agent in the treatment of recurrent ovarian, breast and non-small-cell lung cancers. Traditional dosing regimens with docetaxel involve doses of 60-100 mg/m(2) by infusion every 3 weeks. Now weekly low-dose (30-36 mg/m(2)) regimens are being evaluated in phase I trials. Such low-dose studies require a more sensitive, specific and rapid assay of docetaxel in biological fluids for the determination of pharmacokinetic parameters. Because docetaxel is primarily metabolized by CYP3A4 and is highly protein-bound in the plasma, there is potential for drug drug interactions and high interpatient variability in pharmacokinetics. Therefore, pharmacokinetic studies are an important component to understanding the therapeutic variability of docetaxel-containing chemotherapeutic regimens. METHODS: To this end, we developed an analytical assay for docetaxel based upon tandem LCMS and paclitaxel as an internal standard. The sensitivity of the new assay allowed us to monitor plasma levels of docetaxel out to 48 h after the end of the infusion in patients enrolled in a phase I trial of exisulind (orally, twice daily) receiving weekly docetaxel doses of 30 or 36 mg/m(2) where plasma docetaxel levels are below the lower limit of quantitation for traditional HPLC/UV-based assays at later time-points. RESULTS: The inclusion of the 48-h time-point had significant effects on the calculated pharmacokinetic parameters when using either a three-compartment or non-compartmental analysis. The terminal half-life was significantly increased when the 48-h time-point was included in the pharmacokinetic analysis, and the use of model parameters derived with the inclusion of the 48-h time-point were able to more accurately predict plasma levels at later times. CONCLUSIONS: The results reflect the importance of accurate and sensitive analytical methods for the determination of pharmacokinetic parameters and the effect of this later time-point on docetaxel pharmacokinetic modeling. Further, with the increased use of weekly docetaxel in combination with other agents, the inclusion of these later sampling time-points and sensitive methods for drug level determinations are important components in the description of pharmacokinetic drug interactions. PMID- 12759777 TI - Management of complex perineal injuries. AB - A retrospective study of 25 patients with severe soft tissue injuries to the perineum from the last 14 years was performed to determine the most appropriate management strategy for these problematic wounds. There were 20 (80%) men and 5 (20%) women with an average age of 29 years. Six (24%) patients died of exsanguination from their pelvic injuries within the first few hours of presentation. The remaining 19 patients were taken to the operating room for sigmoidoscopy, diverting colostomy, distal rectal washout, and radical debridement and irrigation of any devitalized tissue. Enteral access by jejunostomy was obtained in six patients. Most patients underwent daily debridement and pulsatile irrigation for at least three days, but usually longer, until the treating surgeon deemed the wound to be clean. None of the patients managed in this fashion developed pelvic sepsis. However, pelvic sepsis did occur in all four patients who did not receive early mandatory daily debridement. Total parenteral nutrition was required in eight patients because there were significant delays in the ability of these patients to resume oral intake. In most previously published series in which daily debridement was not performed, a pelvic sepsis rate of 40% to 80% was reported. Therefore, to optimize the clinical course and recovery from these complex wounds, we conclude that mandatory daily debridement with pulsatile irrigation should be instituted in conjunction with sigmoidoscopy, diversion of the fecal stream, distal rectal irrigation, enteral access, and initial radical debridement of devitalized tissue. PMID- 12759774 TI - Characterization of mutations that are synthetic lethal with pol3-13, a mutated allele of DNA polymerase delta in Saccharomyces cerevisiae. AB - The pol3-13 mutation is located in the C-terminal end of POL3, the gene encoding the catalytic subunit of polymerase delta, and confers thermosensitivity onto the Saccharomyces cerevisiae mutant strain. To get insight about DNA replication control, we performed a genetic screen to identify genes that are synthetic lethal with pol3-13. Mutations in genes encoding the two other subunits of DNA polymerase delta (HYS2, POL32) were identified. Mutations in two recombination genes (RAD50, RAD51) were also identified, confirming that homologous recombination is necessary for pol3-13 mutant strain survival. Other mutations were identified in genes involved in repair and genome stability (MET18/ MMS19), in the control of origin-firing and/or transcription (ABF1, SRB7), in the S/G2 checkpoint (RAD53), in the Ras-cAMP signal transduction pathway (MKS1), in nuclear pore metabolism (SEH1), in protein degradation (DOC1) and in folding (YDJ1). Finally, mutations in three genes of unknown function were isolated (NBP35, DRE2, TAH18). Synthetic lethality between pol3-13 and each of the three mutants pol32, mms19 and doc1 could be suppressed by a rad18 deletion, suggesting an important role of ubiquitination in DNA replication control. We propose that the pol3-13 mutant generates replicative problems that need both homologous recombination and an intact checkpoint machinery to be overcome. PMID- 12759784 TI - Diagnosis of SLAP lesions with Grashey-view arthrography. AB - OBJECTIVE: To examine the accuracy of Grashey views obtained during shoulder arthrography in the diagnosis of clinically relevant superior labrum anterior posterior (SLAP) lesions. DESIGN AND PATIENTS: Grashey views obtained during diagnostic arthrography (conventional and MR) were used to examine the superior labrum. Twenty-eight of 118 shoulder arthrograms obtained during a 27-month period fulfilled study criteria and were correlated for accuracy using arthroscopically confirmed grade 2-4 SLAP lesions as the standard of reference. Arthrograms were graded using the consensus method. Prevalence, sensitivity, specificity, and accuracy were 21%, 50%, 86%, and 79%. The appearance of the superior labrum on the Grashey view was compared subjectively with MR arthrography. Sources of errors were analyzed. CONCLUSION: Grashey views obtained during shoulder arthrography can diagnose clinically relevant SLAP lesions with moderately high specificity, moderate accuracy, and limited sensitivity. Findings on the Grashey view closely resemble those seen on coronal oblique MR arthrography. Grashey views should be considered in patients undergoing shoulder arthrography. PMID- 12759785 TI - Leiomyosarcoma of the distal femur in a patient with a history of bilateral retinoblastoma: a case report and review of the literature. AB - We report a case of primary leiomyosarcoma of the distal femoral shaft arising in a patient who had undergone bilateral orbital enucleation for bilateral retinoblastoma several years previously. Radiography demonstrated an osteolytic, expansive lesion with cortical destruction anteriorly in the distal femoral shaft, and these findings were confirmed on CT. MR imaging revealed an expansive intramedullary lesion with cortical breakthrough and soft tissue extension. The occurrence of a second malignancy in patients with a history of bilateral retinoblastoma is well documented. Many different histological types have been described, with osteosarcoma and leiomyosarcoma occurring with the greatest frequency. PMID- 12759786 TI - Effect of inactivation of poly(hydroxyalkanoates) depolymerase gene on the properties of poly(hydroxyalkanoates) in Pseudomonas resinovorans. AB - The phaZ gene of Pseudomonas resinovorans codes for a poly(hydroxyalkanoates) (PHA) depolymerase. Two phaZ mutants of Pseudomonas resinovorans NRRL B-2649, FOAC001 and FOAC002, were constructed by an in vitro transposition procedure followed by chromosomal integration via homologous recombination. A detailed mapping of the transposon insertion sites and an analysis of the resultant sequences showed that putative fusion polypeptides PhaZ(FOAC001) (239 amino-acid residues) and PhaZ(FOAC002) (85 amino-acid residues) could result from the mutant phaZ genes of FOAC001 and FOAC002, respectively. In vivo PHA degradation data indicated that PhaZ(FOAC001) might still retain a partial PHA depolymerization activity, while PhaZ(FOAC002) is completely devoid of this function. The cell yields and PHA contents of B-2649, FOAC001, and FOAC002 were similar when the cells were grown either under a limiting nitrogen-source (low-N) condition for up to 5 days or in excess N-source (high-N) for 3 days. A dramatic decrease in PHA content was observed in the PhaZ-active B-2649 and FOAC001 cells during prolonged cell growth (5 days) in high-N medium or in response to a shift-up in nitrogen source. The repeat-unit compositions of the PHAs from FOAC001 and FOAC002 contained slightly lower amounts of beta-hydroxyoctanoate and higher beta hydroxytetradecenoate than that of the wild-type B-2649 when grown under a high-N condition. While the molecular masses of the PHAs from FOAC001 and FOAC002 did not vary under any conditions used in this study, those of the wild-type B-2649 were markedly increased in cells either grown for 5 days under a high-N condition or subjected to a nitrogen-source shift-up. These phaZ mutants thus provide a valuable system to study the influence of PHA depolymerase on the accumulation and properties of medium-chain-length PHA. PMID- 12759787 TI - Sequential cloned gene integration in the yeast Kluyveromyces lactis. AB - Two integrating vectors developed for use in Saccharomyces cerevisiae were successfully employed for cloned gene integration in the yeast Kluyveromyces lactis. A delta-integrating vector carrying the dominant selection marker neo allowed tandem integrations of a CUP1p-lacZ cassette into one or two chromosomal sites. A delta/UB-integrating vector, which contains a reusable selection cassette, enabled multiple rounds of integration and the sequential insertion of stable, dispersed copies of CUP1p-lacZ. Subsequent gene expression was closely correlated with integrated copy number illustrating the promise of this method for metabolic engineering in K. lactis. While both vectors contain an S. cerevisiae delta target sequence, the presence of delta-like elements in K. lactis has not been confirmed. Given the degree of illegitimate recombination in this yeast species, the insertions likely occurred at random locations in the chromosomes. PMID- 12759788 TI - Production of verbenol, a high valued food flavourant from a fusant strain of Aspergillus niger. AB - A hyperperformer for the production of verbenol was produced from the fusion of two improved strains of Aspergillus niger. A 2-deoxy glucose de-repressed mutant [high sporulation (50%), viability (80%) showing a conversion of 15.6% of initial alpha-pinene to verbenol in 6 h under the conditions used] was fused with another strain enriched with alpha-pinene (26.4% of alpha pinene converted to verbenol) to obtain a final verbenol conversion yield of 48.6% of initial alpha pinene. PMID- 12759789 TI - Molecular genetics of fungal siderophore biosynthesis and uptake: the role of siderophores in iron uptake and storage. AB - To acquire iron, all species have to overcome the problems of iron insolubility and toxicity. In response to low iron availability in the environment, most fungi excrete ferric iron-specific chelators--siderophores--to mobilize this metal. Siderophore-bound iron is subsequently utilized via the reductive iron assimilatory system or uptake of the siderophore-iron complex. Furthermore, most fungi possess intracellular siderophores as iron storage compounds. Molecular analysis of siderophore biosynthesis was initiated by pioneering studies on the basidiomycete Ustilago maydis, and has progressed recently by characterization of the relevant structural and regulatory genes in the ascomycetes Aspergillus nidulans and Neurospora crassa. In addition, significant advances in the understanding of utilization of siderophore-bound iron have been made recently in the yeasts Saccharomyces cerevisiae and Candida albicans as well as in the filamentous fungus A. nidulans. The present review summarizes molecular details of fungal siderophore biosynthesis and uptake, and the regulatory mechanisms involved in control of the corresponding genes. PMID- 12759790 TI - Bioflavoring and beer refermentation. AB - Various techniques are used to adjust the flavors of foods and beverages to new market demands. Although synthetic flavoring chemicals are still widely used, flavors produced by biological methods (bioflavors) are now more and more requested by consumers, increasingly concerned with health and environmental problems caused by synthetic chemicals. Bioflavors can be extracted from plants or produced with plant cell cultures, microorganisms or isolated enzymes. This Mini-Review paper gives an overview of different systems for the microbial production of natural flavors, either de novo, or starting with selected flavor precursor molecules. Emphasis is put on the bioflavoring of beer and the possibilities offered by beer refermentation processes. The use of flavor precursors in combination with non-conventional or genetically modified yeasts for the production of new products is discussed. PMID- 12759791 TI - Atypical teratoid/rhabdoid tumor of the central nervous system in children: an atypical series and review. AB - Primary atypical teratoid/rhabdoid tumor (AT/RhT) of the central nervous system is a recently described, highly malignant neoplasm in infants and young children. This tumor is an unusual combination of mixed cellular elements, similar but not typical of teratomas, and rhabdoid cells. This tumor is most common in the posterior fossa in children less than 2 years, and is radiologically similar to medulloblastoma. No pathognomonic imaging features are present. The two tumors can be separated on histologic, molecular, and cytogenetic grounds. Separation of these two tumor types is crucial because the prognosis for AT/RhT is grim even with current multimodality treatment. We present four consecutive cases of AT/RhT, three in locations other than the cerebellum, seen at our institution in a 14-month period, indicating that this tumor may be more common than previously thought. PMID- 12759792 TI - Abnormal sagittal sinus blood flow in term infants following a perinatal hypoxic ischaemic insult. AB - Ultrasound is a useful tool for the study of the superior sagittal sinus in neonates. The normal patterns of blood flow have been established, and the technique has been used to diagnose sagittal sinus thrombosis. This report describes the sonographic diagnosis of abnormally sluggish or absent sagittal sinus flow in two term infants. The first was a 2,320-g female infant with postnatal hypoxia. The second was a 5,000-g male infant who developed hypoxic ischaemic encephalopathy after delivery complicated by shoulder dystocia. In both cases the normal pattern of blood flow in the superior sagittal sinus was re established on follow-up ultrasonography. PMID- 12759793 TI - Abdominal aortic injury in a child: intravenous digital subtraction angiogram (IVDSA) for the diagnosis of pediatric vascular trauma. AB - Abdominal aortic injury due to trauma is a rare entity, especially in the pediatric population. We report a 6-year-old girl with partial transection of the abdominal aorta as a result of a motor vehicle accident. The diagnosis was made with IVDSA. The patient survived the injury. We discuss the imaging findings, mechanisms, and associated injuries of abdominal aortic trauma in children. PMID- 12759794 TI - Prescribing information in 26 countries: a comparative study. AB - This study was set up to document the variability of prescribing information from different sources concerning indications, side effects and cautions of selected drugs. An original method to measure the degree of information agreement among different written materials, such as summaries of product characteristics, package inserts and data sheets, and a widely accepted reference text was developed. The results show that there is substantial disagreement in the materials available to prescribers and patients in different countries. Disagreement was even found within a single country when written materials from different brands of the same drug were compared. The discordance can be explained by the fact that the evidence available for each drug is considered/assessed differently by separate countries. It is argued that the discrepancies found may mislead prescribers, patients and those comparing drug-use patterns across countries. National regulatory authorities have a key role to play in remedying this situation, and a two-pronged approach is proposed. At the international level, national authorities should strengthen collaboration and information interchange and, at the national level, should implement appropriate measures aimed at removing contradictory statements on drug-information materials that have no reason to be different. Finally, further training and continued education aimed at drug regulatory officials could provide the necessary knowledge and enable national authorities to meet the need for drug information that is independent of commercial interests. PMID- 12759795 TI - Cost-effectiveness analysis of the first-line therapies for nicotine dependence. AB - BACKGROUND: Nicotine dependence is the major obstacle for smokers who want to quit. Guidelines have identified five effective first-line therapies, four nicotine replacement therapies (NRTs)--gum, patch, nasal spray and inhaler--and bupropion. Studying the extent to which these various treatments are cost effective requires additional research. OBJECTIVES: To determine cost effectiveness (CE) ratios of pharmacotherapies for nicotine dependence provided by general practitioners (GPs) during routine visits as an adjunct to cessation counselling. METHODS: We used a Markov model to generate two cohorts of one-pack a-day smokers: (1) the reference cohort received only cessation counselling from a GP during routine office visits; (2) the second cohort received the same counselling plus an offer to use a pharmacological treatment to help them quit smoking. The effectiveness of adjunctive therapy was expressed in terms of the resultant differential in mortality rate between the two cohorts. Data on the effectiveness of therapies came from meta-analyses, and we used odds ratio for quitting as the measure of effectiveness. The costs of pharmacotherapies were based on the cost of the additional time spent by GPs offering, prescribing and following-up treatment, and on the retail prices of the therapies. We used the third-party-payer perspective. Results are expressed as the incremental cost per life-year saved. RESULTS: The cost per life-year saved for only counselling ranged from Euro 385 to Euro 622 for men and from Euro 468 to Euro 796 for women. The CE ratios for the five pharmacological treatments varied from Euro 1768 to Euro 6879 for men, and from Euro 2146 to Euro 8799 for women. Significant variations in CE ratios among the five treatments were primarily due to differences in retail prices. The most cost-effective treatments were bupropion and the patch, and, then, in descending order, the spray, the inhaler and, lastly, gum. Differences in CE between men and women across treatments were due to the shape of their respective mortality curve. The lowest CE ratio in men was for the 45- to 49-year-old group and for women in the 50- to 54-year-old group. Sensitivity analysis showed that changes in treatment efficacy produced effects only for less-well proven treatments (spray, inhaler, and bupropion) and revealed a strong influence of the discount rate and natural quit rate on the CE of pharmacological treatments. CONCLUSION: The CE of first-line treatments for nicotine dependence varied widely with age and sex and was sensitive to the assumption for the natural quit rate. Bupropion and the nicotine patch were the two most cost-effective treatments. PMID- 12759796 TI - Impaired postural compensation for respiration in people with recurrent low back pain. AB - This study evaluated the degree to which the disturbance to posture from respiration is compensated for in healthy normals and whether this is different in people with recurrent low back pain (LBP), and to compare the changes when respiratory demand is increased. Angular displacement of the lumbar spine and hips, and motion of the centre of pressure (COP), were recorded with high resolution and respiratory phase was recorded from ribcage motion. With subjects standing in a relaxed posture, recordings were made during quiet breathing, while breathing with increased dead-space to induce hypercapnoea, and while subjects voluntarily increased their respiration to match ribcage expansion that was induced in the hypercapnoea condition. The relationship between respiration and the movement parameters was measured from the coherence between breathing and COP and angular motion at the frequency of respiration, and from averages triggered from the respiratory data. Small angular changes in the lumbopelvic and hip angles were evident at the frequency of respiration in both groups. However, in quiet standing, the LBP subjects had a greater displacement of their COP that was associated with respiration than the control subjects. The LBP group had a trend for less hip motion. There were no changes in the movement parameters when respiratory demand increased involuntarily via hypercapnoea, but when respiration increased voluntarily, the amplitude of motion and the displacement of the COP increased in both groups. The present data suggest that the postural compensation to respiration counteracts at least part of the disturbance to posture caused by respiration and that this compensation may be less effective in people with LBP. PMID- 12759797 TI - Carbohydrate and alditol analysis by high-performance anion-exchange chromatography coupled with electrochemical detection at a cobalt-modified electrode. AB - A cobalt oxyhydroxide film dispersed on a carbon electrode surface was characterized and proposed as an amperometric sensor for determination of alditols and carbohydrates in flowing streams. Complex mixtures of carbohydrates were separated by anion-exchange chromatography using a moderately alkaline solution as mobile phase. The cobalt modified electrode (GC-Co) was employed under a constant applied potential of 0.5 V (vs Ag/AgCl). Under these experimental conditions the detection limits (S/N=3) for all analyzed electroactive molecules ranged between 0.3 micromol L(-1) and 1.5 micromol L(-1) and the dynamic linear ranges spanned generally three orders of magnitude above the relevant detection limits. Analytical determinations of carbohydrates and alditols in red and white wines, are reported. PMID- 12759799 TI - Speciation of Co(II), Co(III), and Cu(II) in ethylenediamine solutions by capillary electrophoresis. AB - A capillary electrophoretic (CE) method for the speciation of Co(II), Co(III), and Cu(II) in electroless copper-plating baths containing ethylenediamine (En) has been developed. The method is based on the selective pre-capillary derivatization of Co(II) with 1,10-phenanthroline (Phen) followed by CE separation of stable [CoPhen(3)](2+), [CoEn(3)](3+), and [CuEn(2)](2+) chelates. The proposed derivatization procedure protects Co(II) from oxidation by dissolved oxygen and enables rapid determination of all three metal species within a single run. The optimized separations were carried out in a fused silica capillary (57 cmx75-microm I.D.) filled with an ethylenediamine sulfate electrolyte (20 mmol L( 1) H(2)SO(4), pH 7.0 with En, applied voltage +30 kV) using direct UV detection at 214 nm. The detection limits for a signal-to-noise ratio of 3 and 10 s, hydrodynamic injections were 5x10(-6) mol L(-1) for Cu(II), 1x10(-6) mol L(-1) for Co(III), and 4x10(-7) mol L(-1) for Co(II). Application of the method to the speciation of Co(II), Co(III), and Cu(II) in copper-plating bath samples is also demonstrated. PMID- 12759798 TI - Differential-pulse polarography (DPP) determination of betamethasone valerate in dosage form. AB - The electrochemical reduction of betamethasone valerate (BV) in a pharmaceutical formulation containing neomycin has been carried out in Britton-Robinson buffer (BRB) (0.04 mol L(-1)) by differential-pulse polarography (DPP). BV exhibits a well-defined irreversible reduction peak at -1.03 V/ref. The influence of pH on the reduction of BV was studied in Britton-Robinson buffer (pH range 1.7-10). A method for the analysis of BV in BRB (0.04 mol L(-1)), which allows quantification over the range 3.9x10(-6)-1.1x10(-4) mol L(-1), was proposed and successfully applied to the determination of BV in tablets with mean recovery and relative standard deviation of 100.81% and 0.45%, respectively. PMID- 12759800 TI - Instrumentation for FT-IR reflection spectroscopy with synchrotron radiation. AB - A versatile experimental set-up for infrared reflectance measurements with synchrotron radiation and its adaptation to a beamline is presented. Particular consideration is given to the collimation and the polarization of incident radiation. The performance is characterized with experimental results. Due to the high brilliance of the synchrotron radiation source, the irradiation of samples smaller than 1 mm(2) was found to be improved by more than one order of magnitude when compared to a globar. PMID- 12759801 TI - Altered response to tryptophan supplementation after long-term abstention from MDMA (ecstasy) is highly correlated with human memory function. AB - RATIONALE: MDMA (ecstasy; +3,4-methylenedioxymethamphetamine) damages brain serotonin (5-HT) neurons and, in non-human primates, a loss of various 5-HT axonal markers persists for several years. This raises the question of whether long lasting effects occur in human beings that persist even after they have stopped using MDMA. OBJECTIVES: We therefore assessed the effects of an indirect 5-HT manipulation on functions thought to be affected by MDMA use in people who had stopped using MDMA (ex-users) compared with continuing users and non-users. METHODS: Ninety-six participants were recruited: 32 ex-users who had stopped using MDMA for >1 year (mean, 2.4 years); 32 current users and 32 polydrug controls who had never used MDMA but were matched with ex-users and controls on cannabis use and pre-morbid IQ. Participants were given an amino acid mixture that contained either no tryptophan (T-) or augmented tryptophan (T+) and assessed before and 5 h after the drink on measures of cognitive function and mood. RESULTS: T+ and T- produced plasma tryptophan augmentation and depletion, respectively, in all three groups. Ex-users' plasma tryptophan levels in response to T+ were significantly higher than other groups. Ex-users' performance on a delayed prose recall task improved after T+ and lessened after T-. Changes in ex users' free plasma tryptophan levels correlated highly (r=-0.9) with their baseline performance on immediate and delayed prose recall; change in total plasma tryptophan correlated (r=-0.81) with delayed recall. Further, total baseline plasma tryptophan correlated with number of years they had used MDMA before quitting. Baseline differences between groups were found on learning, working memory, aggression and impulsivity. T- did not produce differential effects in the three groups. CONCLUSIONS: Our results suggest that prolonged abstinence from MDMA might be associated with altered tryptophan metabolism. Ex users showing the poorest memory function at baseline were also those who metabolised least tryptophan. These findings may reflect pre-morbid differences in 5-HT function of those who stop using this drug or consequences of MDMA use that emerge after abstention. Aggression is also associated with MDMA use and subsequent abstinence. PMID- 12759802 TI - Source monitoring improvement in patients with schizophrenia receiving antipsychotic medications. AB - RATIONALE: The absence of a relationship between cognitive deficit treatment response and positive symptom treatment response is often assumed, and few data have shed light on this issue. Most of these data have been collected using standard neuropsychological measures, which are ill-designed to assess the types of neurocognitive disturbances associated with psychotic symptoms. This study investigates the effect of treatment on source monitoring performance and its relation to the reduction of certain psychotic symptoms associated with the inability to identify self-generated mental events, known as "autonoetic agnosia". OBJECTIVES: To determine whether risperidone, olanzapine, and haloperidol were differentially effective in reducing autonoetic agnosia and whether changes in this aspect of cognition were related to reduction of specific symptoms of psychosis. METHODS: From a cohort of 49 patients diagnosed with schizophrenia by DSM-IV criteria and randomly assigned to double-blind treatment with risperidone, olanzapine, or haloperidol, 16 patients were identified with symptoms believed to reflect autonoetic agnosia ("target symptoms") as assessed with the Schneiderian Symptom Rating Scale, and then evaluated during a baseline period, and then at 1, 2, and 3 weeks. Autonoetic agnosia was assessed as the ability of a patient to distinguish self-generated words from both experimenter generated words and pictorially presented words. RESULTS: Analysis of patients from all treatment groups found a significant reduction in the number of "target" Schneiderian symptoms. Discrimination for items from the self-generated and heard sources significantly improved with treatment, as did the number of self generated items that patients remembered as coming from the heard source ("self hear errors"). The correlation between improvement in recognition of self generated items and reduction in target Schneiderian symptoms after 2 weeks of treatment suggested a modest relationship between symptom improvement and changes in autonoetic agnosia. CONCLUSIONS: While the differences between medications were not statistically significant, antipsychotic medication in general was associated with improvements in symptoms and cognitive deficits that may underlie autonoetic agnosia. Improvement of autonoetic agnosia was a weak predictor of positive symptom improvement in a limited sample. PMID- 12759805 TI - DMXB, an alpha7 nicotinic agonist, normalizes auditory gating in isolation-reared rats. AB - RATIONALE: Impaired auditory gating is common in schizophrenic patients. Evidence suggests that this deficit is related to a reduced number of alpha(7) nicotinic receptors and therefore treatment with alpha(7) nicotinic agonists may improve this condition. 3-(2,4)-Dimethoxybenzylidine anabaseine (DMXB; also known as GTS 21) is such an agonist and has shown efficacy in mice both orally and intraperitoneally. OBJECTIVE: Rats reared in social isolation post weaning have demonstrated a deficit in auditory gating similar to that seen in schizophrenia patients. The current study determined the effects of DMXB on auditory gating in awake, freely moving rats, comparing a group born and raised in-house and reared in isolation post-weaning (isolation reared) with a group shipped from the supplier as adults and housed in groups prior to surgery (controls). METHODS: Ten unmedicated, baseline recordings were obtained following surgical implantation of a recording electrode. All control group rats and the isolation-reared rats that showed deficient gating at baseline were treated with 1.0, 3.33, 10 or 33 mg/kg DMXB, IP, to determine the drug's impact on auditory gating. RESULTS: Isolation reared rats had significantly improved auditory gating at the 3.33, 10 and 33 mg/kg doses, while control rats had a significant impairment in their auditory gating at the 33 mg/kg dose. CONCLUSIONS: DMXB improved the auditory gating deficit seen in isolation-reared rats. As previously observed in another model, the change was produced through a decrease in the test amplitude in isolation reared animals. Control animals had a significant reduction in conditioning amplitude at the high dose, which produced the loss of auditory gating. The results in the isolation-reared rats are in concert with previous studies which found similar improvement in auditory gating following administration of DMXB to DBA mice, the only differences being in the duration of the effect. PMID- 12759806 TI - Opioids, cocaine, and food change runtime distribution in a rat runway procedure. AB - RATIONALE: The nature of the relationship between the dose of a drug of abuse and its reinforcing effect has come under close scrutiny. It is currently debated if the steep ascending part of the biphasic dose-response curve typically obtained in multiple-dosing lever-press-based operant conditioning procedures represents a satiety-driven, all-or-none response or if the response is gradual and tightly adjusted to the various doses of the reinforcer. OBJECTIVES: Dose-response relationships of drug reinforcers (remifentanil, alfentanil, morphine, cocaine) as well as a physiological reinforcer, i.e. food (sweetened condensed milk) were investigated in a different operant conditioning paradigm, i.e. a runway procedure. METHODS: Administration of the mu opioid receptor agonists remifentanil (0.0032-0.1 mg/kg IV), alfentanil (0.032 mg/kg IV), morphine (0.032 1 mg/kg IV), the psychostimulant cocaine (0.001-0.1 mg/kg IV) or sweetened condensed milk (diluted 1:100-1:3 in water) was made contingent upon alley running for male Sprague-Dawley rats. RESULTS: All drug reinforcers dose dependently decreased runtimes and, more importantly, significantly increased the percentage of runs at a certain speed (> or =10 cm/s) from an average 29% to an average 71% (water versus milk, 22% versus 83%). CONCLUSION: Both drug- and food reinforcers produced a discontinuous, qualitative change in the rats' operant behavior rather than a simple gradual increase along a continuum, an effect that could be seen clearly only after a histogram analysis of runtime distribution. PMID- 12759807 TI - A microdialysis profile of beta-endorphin and catecholamines in the rat nucleus accumbens following alcohol administration. AB - RATIONALE: Alcohol stimulates the release of dopamine in the nucleus accumbens (NACB) of rats, mice and humans. There is evidence to suggest that the activation of beta-endorphin (beta-EP) in the mesolimbic pathway by alcohol and other drugs of abuse may be associated with the rise in dopamine levels in the NACB. OBJECTIVES: The present studies investigate whether the release of beta-EP in the NACB is (1) dependent on the dose of alcohol that is administered, and (2) associated with changes in the extracellular concentrations of the catecholamines dopamine and norepinephrine, and the dopamine metabolites 3,4 dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), in the NACB. METHODS: Male Sprague-Dawley rats were implanted with a microdialysis probe positioned in the shell region of the NACB. Artificial cerebrospinal fluid was pumped at a rate of 2.3 microl/min in awake and freely moving animals and the dialysate was collected at 30-min intervals. After a baseline period, rats were injected intraperitoneally with either physiological saline or one of three doses of alcohol: 0.8, 1.6, or 2.4 g ethanol/kg body weight. The dialysates collected were analyzed with radioimmunoassay, to estimate the content of beta-EP; and high performance liquid chromatography, to estimate the content of dopamine, norepinephrine, DOPAC and HVA. RESULTS: Alcohol induced a dose-dependent increase in the extracellular levels of beta-EP and dopamine. However, elevations in the extracellular levels of norepinephrine, DOPAC and HVA did not reach significance. The largest increase in beta-EP and dopamine was observed with the 2.4 g/kg dose. CONCLUSION: The alcohol-induced release of beta-EP and dopamine in the NACB is dose-dependent, where the highest dose resulted in more pronounced concentrations in the dialysate. Furthermore, the increase in the extracellular levels of dopamine appeared to occur at an earlier time point following alcohol administration, than for beta-EP. These results suggest that alcohol stimulates dopamine and beta-EP in the NACB, but probably does so via independent mechanisms. PMID- 12759808 TI - Expectations and placebo responses to caffeine-associated stimuli. AB - RATIONALE: To test the theory that expectations control placebo responses. OBJECTIVE: Subjects (n=20) were asked how much they expected their arousal to increase after one or two cups of coffee, and were subsequently exposed to one or two cups of decaffeinated coffee, or to caffeine equivalent to one or two cups of coffee (200 and 400 mg). The expectancy theory of placebo responses predicts a positive correlation between expectations and actual placebo responses. METHODS: Dependent variables were acoustic startle eyeblink and skin conductance responses, blood pressure and heart rate, and measures of subjective arousal. RESULTS: Caffeine increased startle eyeblink and skin conductance responses, as well as blood pressure and subjective arousal. Decaffeinated coffee increased startle eyeblink and skin conductance responses, but had no effect on subjective arousal, although the participants clearly expected increased subjective arousal after both one and two cups of coffee. However, there were significant correlations between the alertness expected after coffee, and the actual alertness recorded after decaffeinated coffee. CONCLUSIONS: The main finding in this study was that relatively strong expectations about the effects of coffee did not generate placebo responses after administration of decaffeinated coffee. Expectations were dose dependent, whereas the placebo response was not. However, expected alertness after coffee predicted recorded alertness after coffee. In sum, the expectancy theory of placebo effects received only limited support. PMID- 12759809 TI - Biotransformation of beta-ketosulfides to produce chiral beta-hydroxysulfoxides. AB - The biotransformations of a series of substituted phenylthio-2-propanone and benzylthio-2-propanone were carried out using Helminthosporium sp. NRRL 4671, Mortierella isabellina ATCC 42613, or Rhodococcus erythropolis IGTS8. Several products gave microbial oxidation of sulfide to sulfoxide and reduction of carbonyl to secondary alcohol, producing beta-hydroxysulfoxides in medium to high enantiomeric and diastereomeric purities. Fungal biotransformations using Helminthosporium sp. and M. isabellina resulted in the opposite sulfoxide configurations of various beta-hydroxysulfoxide products. PMID- 12759810 TI - Heterologous production of flavanones in Escherichia coli: potential for combinatorial biosynthesis of flavonoids in bacteria. AB - Chalcones, the central precursor of flavonoids, are synthesized exclusively in plants from tyrosine and phenylalanine via the sequential reaction of phenylalanine ammonia-lyase (PAL), cinnamate-4-hydroxylase (C4H), 4 coumarate:coenzyme A ligase (4CL) and chalcone synthase (CHS). Chalcones are converted into the corresponding flavanones by the action of chalcone isomerase (CHI), or non-enzymatically under alkaline conditions. PAL from the yeast Rhodotorula rubra, 4CL from an actinomycete Streptomyces coelicolor A3(2), and CHS from a licorice plant Glycyrrhiza echinata, assembled as artificial gene clusters in different organizations, were used for fermentation production of flavanones in Escherichia coli. Because the bacterial 4CL enzyme attaches CoA to both cinnamic acid and 4-coumaric acid, the designed biosynthetic pathway bypassed the C4H step. E. coli carrying one of the designed gene clusters produced about 450 microg naringenin/l from tyrosine and 750 microg pinocembrin/l from phenylalanine. The successful production of plant-specific flavanones in bacteria demonstrates the usefulness of combinatorial biosynthesis approaches not only for the production of various compounds of plant and animal origin but also for the construction of libraries of "unnatural" natural compounds. PMID- 12759811 TI - Calcifying pseudo-tumor of the spine: description of a case and review of the literature. AB - A case of calcifying pseudo-tumor of the thoracic spine, a rare lesion with tumor like behavior and a probable inflammatory-reactive origin, is described. The clinical-pathological and neuro-radiological aspects of this lesion are discussed in relation to surgical treatment. In accordance with the other cases reported in the literature, the case observed confirmed the benign behavior of the lesion and the effectiveness of surgical treatment for achieving complete resolution of clinical symptoms without any recurrences, even when removal is only subtotal. PMID- 12759812 TI - The distinction between Liddle syndrome and apparent mineralocorticoid excess. PMID- 12759813 TI - Source partitioning using stable isotopes: coping with too many sources. AB - Stable isotopes are increasingly being used as tracers in environmental studies. One application is to use isotopic ratios to quantitatively determine the proportional contribution of several sources to a mixture, such as the proportion of various pollution sources in a waste stream. In general, the proportional contributions of n+1 different sources can be uniquely determined by the use of n different isotope system tracers (e.g., delta13C, delta15N, delta18O) with linear mixing models based on mass balance equations. Often, however, the number of potential sources exceeds n+1, which prevents finding a unique solution of source proportions. What can be done in these situations? While no definitive solution exists, we propose a method that is informative in determining bounds for the contributions of each source. In this method, all possible combinations of each source contribution (0-100%) are examined in small increments (e.g., 1%). Combinations that sum to the observed mixture isotopic signatures within a small tolerance (e.g., +/-0.1 per thousand ) are considered to be feasible solutions, from which the frequency and range of potential source contributions can be determined. To avoid misrepresenting the results, users of this procedure should report the distribution of feasible solutions rather than focusing on a single value such as the mean. We applied this method to a variety of environmental studies in which stable isotope tracers were used to quantify the relative magnitude of multiple sources, including (1) plant water use, (2) geochemistry, (3) air pollution, and (4) dietary analysis. This method gives the range of isotopically determined source contributions; additional non-isotopic constraints specific to each study may be used to further restrict this range. The breadth of the isotopically determined ranges depends on the geometry of the mixing space and the similarity of source and mixture isotopic signatures. A sensitivity analysis indicated that the estimated ranges vary only modestly with different choices of source increment and mass balance tolerance parameter values. A computer program (IsoSource) to perform these calculations for user-specified data is available at http://www.epa.gov/wed/pages/models.htm. PMID- 12759814 TI - Superparasitism and sex ratio adjustment in a wasp parasitoid: results at variance with Local Mate Competition? AB - Anaphes nitens is a solitary parasitoid of the egg capsules of the Eucalyptus snout beetle, Gonipterus scutellatus. Some traits of its natural history suggest that Local Mate Competition (LMC) could account for sex ratio adjustment in this species. We tested whether males emerged early, a prerequisite for fully local mating, and investigated the occurrence and effect of superparasitism on adult size and pre-emergence mortality, factors that might influence sex ratio adjustment. We found in field-collected egg capsules that males emerged first. To investigate the effects of superparasitism on adult size, we compared the sizes of parasitoids that emerged early and late from egg capsules collected in the field, and from egg capsules parasitized and superparasitized in the laboratory. Superparasitism reduced parasitoid size, affecting females more strongly than males, and increased pre-emergence mortality. We estimated A. nitens sex ratio and parasitism rate in the field during 2 years in five localities and during 4 years in a sixth. Following LMC we expected an increase in sex ratio (proportion of males) with increasing parasitism rate (assumed to reflect parasitoid density). We found that sex ratio decreased from 0.38 when the parasitism rate was low (0-20%) to 0.21 when parasitism was high (80-100%). In contrast with field results, a laboratory experiment showed that: (1) at a low parasitism level sex ratio was clearly female biased (0.28+/-0.04), (2) at a high parasitism level sex ratio increased (0.40+/-0.07), (3) male larval survivorship was not lower than female survivorship, and (4) low-quality hosts (i.e. superparasitized) were allocated more males. We conclude that LMC cannot explain the sex ratio adjustment observed in the field, even at low parasitism rates, and alternative explications for highly female-biased sex ratios must be found. One such alternative is female-biased dispersal. PMID- 12759815 TI - Intraspecific variation in palatability and defensive chemistry of brown seaweeds: effects on herbivore fitness. AB - When offered a choice between brown seaweeds (Phaeophyta) from shallow inshore populations versus deeper offshore populations along the mid-Atlantic coast of the United States of America, the herbivorous amphipod Ampithoe longimana consistently preferred plants from the inshore populations. This was the case for three species (Dictyota menstrualis, Spatoglossum schroederi, and Sargassum filipendula) collected from each of a single inshore and offshore site, and for one species (D. menstrualis) collected from each of three inshore and three offshore sites. Bioassay-guided fractionation of chemical crude extracts from D. menstrualis suggested that the relative unpalatability of the offshore plants was due to the lipid-soluble secondary metabolites 4beta-hydroxydictyodial A and 18, O-dihydro-4beta-hydroxydictyodial A 18-acetate, along with minor compounds that were not fully identified. The inshore-offshore pattern did not appear to result from induction of defenses due to herbivory by mesograzers, as mesograzer densities were higher on the more palatable inshore plants. Herbivore feeding preferences for inshore versus offshore seaweeds matched the effects of those seaweeds on their fitness. When juvenile amphipods were raised on inshore versus offshore tissues of D. menstrualis, amphipod survivorship, growth, and ovulation were significantly suppressed on the offshore compared to the inshore tissues. Few previous investigations have studied intraspecific variance in seaweed palatability. We extend these by showing that between-population differences in palatability can persist for several years and by demonstrating that this variance is chemically based and has dramatic effects on herbivore fitness. PMID- 12759816 TI - Visualization of gene expression of prolactin-receptor (PRL-R) by in situ hybridization in reproductive organs of Typhlonectes compressicauda, a gymnophionan amphibian. AB - The expression of the long form of prolactin receptor (PRL-R) mRNAs was studied in reproductive organs from both male and female Typhlonectes compressicauda, an amphibian, by quantitative in situ hybridization. These PRL-R mRNAs were expressed in all the organs studied. In ovarian tissue, mRNAs coding for the long form of PRL-R were strongly expressed in ovaries with compact and vascularized corpora lutea, i.e., during the middle and the end of pregnancy. During the other periods of cycle, sexual rest, vitellogenesis and beginning of pregnancy, expression of these mRNAs was lower than in other periods. In the testis, mRNAs coding for the long form of PRL-R were expressed in germ cells as well as in Leydig and Sertoli cells. In mullerian glands, important variations of expression in these mRNAs have been observed with a large increase during the breeding period and a decrease during sexual rest. Variations of gene expression of the long form of PRL-R for all tissues studied are correlated to synthesis of PRL in the pituitary in both male and female Typhlonectes compressicauda. PMID- 12759817 TI - The origins and genetic structure of three co-resident Chinese Muslim populations: the Salar, Bo'an and Dongxiang. AB - A genome-based investigation of three Muslim populations, the Salar, Bo'an, and Dongxiang, was conducted on 212 individuals (148 males, 64 females) co-resident in Jishisan County, a minority autonomous region located in the province of Gansu, PR China. The Salar are believed to be of Turkic origin, whereas the Bo'an and Dongxiang both speak Mongolian. Biparental dinucleotide markers on chromosomes 13 and 15 indicated elevated mean homozygosity in the Salar (0.32), Bo'an (0.32), and Dongxiang (0.27), equivalent to inbreeding coefficients ( F(is) ) of 0.16; 0.12; 0.01, confirming varying levels of endogamous and consanguineous marriage in all three communities. Y-chromosome unique event polymorphisms (UEPs) showed that males in the three communities shared common ancient origins, with 80 90% of haplotypes in common. However, the high levels of community-specific Y chromosome STR haplotypes strongly suggested the action(s) of founder effect, genetic drift and preferential consanguinity during more recent historical time. By comparison with the marked inter-community differentiation revealed by the Y chromosome STRs (29.4%), the mtDNA data indicated similarity between the female lineages of each community with just 1.2% inter-community variation. The combined use of these different marker systems gives an in-depth historical perspective, and provides evidence of past inter-marriage between genetically diverse male founders of each community and Han Chinese females with subsequent community endogamy. PMID- 12759820 TI - Relationships between cognitive and behavioral measures of executive function in children with brain disease. AB - This study addressed the clinical and construct validity of the Behavior Rating Inventory of Executive Function. (BRIEF: Gioia, Isquith, Guy, & Kenworthy, 2000), a questionnaire designed to tap behavioral aspects of executive functions in children. BRIEF profiles in early treated phenylketonuria (PKU; n=44), early treated hydrocephalus (n=45), frontal focal lesions (n=20) and controls (n=80) were examined. Clinical validity was supported through significant between-group comparisons, especially between the frontal focal lesion group and other groups. To examine construct validity, raw scores on cognitive executive function measures including the Contingency Naming Test (CNT), Rey Complex Figure (RCF), Tower of London (TOL), and Controlled Oral Word Association Test (COWAT), were correlated with BRIEF scale scores. Few significant correlations were found, indicating cognitive and behavioral measures appear to tap different constructs within the executive function domain. A dissociation was found between behavioral and cognitive impairments in the frontal as opposed to PKU and hydrocephalus groups. This is discussed in relation to underlying pathology, the cognitive measures used, and possible limitations in the BRIEF's usefulness for measuring behavioral executive dysfunction in groups only mildly affected by neurological compromise. PMID- 12759818 TI - Sequence variation in the CHAT locus shows no association with late-onset Alzheimer's disease. AB - There is substantial evidence for a susceptibility gene for late-onset Alzheimer's disease (AD) on chromosome 10. One of the characteristic features of AD is the degeneration and dysfunction of the cholinergic system. The genes encoding choline acetyltransferase (ChAT) and its vesicular transporter (VAChT), CHAT and SLC18A3 respectively, map to the linked region of chromosome 10 and are therefore both positional and obvious functional candidate genes for late-onset AD. We have screened both genes for sequence variants and investigated each for association with late-onset AD in up to 500 late-onset AD cases and 500 control DNAs collected in the UK. We detected a total of 17 sequence variants. Of these, 14 were in CHAT, comprising three non-synonymous variants (D7N in the S exon, A120T in exon 5 and L243F in exon 8), one synonymous change (H547H), nine single nucleotide polymorphisms in intronic, untranslated or promoter regions, and a variable number of tandem repeats in intron 7. Three non-coding SNPs were detected in SLC18A3. None demonstrated any reproducible association with late onset AD in our samples. Levels of linkage disequilibrium were generally low across the CHAT locus but two of the coding variants, D7N and A120T, proved to be in complete linkage disequilibrium. PMID- 12759821 TI - Adaptive skills and executive function in autism spectrum disorders. AB - There is active debate regarding the nature of executive dysfunction in autism. Additionally, investigations have yet to show a relationship between deficits in executive function and the everyday behavioral difficulties that may originate from them. The present study examined the relationship between executive abilities and adaptive behavior in 35 children with Autism Spectrum Disorders, using two parent reports of everyday functioning, the Vineland Adaptive Behavior Scales (VABS) and the Behavior Rating Inventory of Executive Function (BRIEF). Results found several relationships: The Initiate and Working Memory domains were negatively correlated with most domains of adaptive behavior. Also, the Communication and Socialization domains of the VABS were negatively correlated with several areas of executive functioning, suggesting that impairments in executive abilities are strongly associated with the deficits in communication, play and social relationships found in children with autism. PMID- 12759822 TI - Confirmatory factor analysis of the Behavior Rating Inventory of Executive Function (BRIEF) in a clinical sample. AB - Evidence for the validity of the Behavior Rating Inventory of Executive Function (BRIEF; Gioia, Isquith, Guy, & Kenworthy, 2000) based on internal structure was examined in a sample of children with mixed clinical diagnoses via maximum likelihood confirmatory factor analysis. Four alternative factor models of children's executive function, based on current theories that posit a unidimensional versus fractionated model (Rabbitt, 1997; Shallice & Burgess, 1991), using the revised 9-scale BRIEF configuration that separates two components of the Monitor scale, were examined for model fit. A 3-factor structure best modeled the data when compared directly with 1-, 2-, and 4-factor models. The 3-factor model was defined by a Behavior Regulation factor consisting of the BRIEF Inhibit and Self-Monitor scales, an Emotional Regulation factor consisting of the Emotional Control and Shift scales, and a Metacognition factor composed of the Working Memory, Initiate, Plan/Organize, Organization of Materials, and Task-Monitor scales. The findings support a fractionated, multi component view of executive function as measured by the BRIEF. PMID- 12759823 TI - Parent and self-report ratings of executive function in adolescents with myelomeningocele and hydrocephalus. AB - Parent and self-report ratings were obtained for 28 adolescents with myelomeningocele and congenital hydrocephalus using the Behavior Assessment System for Children (BASC; Reynolds & Kamphaus, 1998a, 1998b) and the Behavior Rating Inventory of Executive Function (BRIEF; Gioia, Isquith, Guy, & Kenworthy, 2000; Guy, Gioia, & Isquith, 1998). Parents rated their children as having significantly more problems, compared to published norms, on the BRIEF but not the BASC. Adolescents rated themselves as having significantly more problems, compared to parent ratings, on scales comprising the BRIEF Behavioral Regulation Index; but not on common scales of the BASC. Parents also reported more problems on the BRIEF Global Executive Composite than on all three primary BASC indices, and more problems on the BRIEF Metacognition Index than on the BRIEF Behavioral Regulation Index. We conclude that the BRIEF captures salient executive dysfunction among individuals with myelomeningocele and hydrocephalus, and may be useful in identifying needs for intervention that might not be identified by broad-band behavior rating scales alone. PMID- 12759824 TI - Long-term executive function deficits in children with traumatic brain injuries: assessment using the Behavior Rating Inventory of Executive Function (BRIEF). AB - Long-term deficits in executive functions following childhood traumatic brain injuries (TBI) were examined using the Behavior Rating Inventory of Executive Function (BRIEF). Parents completed the BRIEF approximately 5 years postinjury as part of a prospective study of children injured between the ages of 6 and 12. The children were between 10 and 19 years of age at the time of the assessment, and included 33 with severe TBI, 31 with moderate TBI, and 34 with orthopedic injuries. Parents also rated children's adaptive functioning and completed several other measures of parent and family functioning. Children were administered a neuropsychological test battery that included several measures of executive functions. The groups displayed a significant linear trend in BRIEF scores, with the largest deficits in executive functions reported in children with severe TBI. BRIEF scores were related consistently across groups to a test of working memory, but not to other neuropsychological measures. BRIEF scores also predicted children's adaptive functioning and behavioral adjustment, as well as parent psychological distress, perceived family burden, and general family functioning. The findings indicate that TBI results in long-term deficits in executive functions that are related to children's psychosocial outcomes, as well as to parent and family functioning. PMID- 12759825 TI - Parental reports of executive dysfunction in adolescents with bipolar disorder. AB - Bipolar disorder (BPD) is a serious mental illness that affects children and adolescents at a rate similar to that seen in adults. Extremely little is known, however, about cognitive functioning in childhood and adolescent BPD. The present study represents an initial effort to examine executive functioning in adolescents with BPD who are in a manic or mixed mood state, by collecting data from caregivers about the participants' performance on everyday tasks thought to be mediated by executive functioning abilities, using the Behavior Rating Inventory of Executive Function. In comparison to healthy volunteers, adolescents with BPD exhibited significant elevations across all of the measured functional domains. These elevations were evident even in adolescents with BPD who did not have comorbid ADHD, although they were most prominent in those with comorbidity. The findings suggest that adolescents with BPD have functional deficits on tasks requiring executive functioning skills that are not explicable solely on the basis of comorbid ADHD. PMID- 12759826 TI - The relationship between parental report on the BRIEF and performance-based measures of executive function in children with moderate to severe traumatic brain injury. AB - The Behavior Rating Inventory of Executive Function (BRIEF) is a questionnaire that assesses parental observations of behaviors associated with executive function in children in the home environment. The current investigation examines the relationship between the BRIEF and individually-administered neuropsychological tests in children with traumatic brain injury. Forty-eight children with moderate to severe traumatic brain injury were administered the WISC-III and several performance-based tests of executive function (the Wisconsin Card Sorting Test, Trail Making Test Part B, verbal fluency), and a parent completed the BRIEF. Results indicate that the Metacognition Index from the BRIEF correlates with Verbal IQ, but none of the index scores from the BRIEF correlate with any of the performance-based tests of executive function. Results are discussed with respect to the ecological validity of standardized clinical neuropsychological tests of executive function. PMID- 12759827 TI - The Behavior Rating Inventory of Executive Function: commentary. PMID- 12759831 TI - Congenital hypothyroidism: an analysis of persisting deficits and associated factors. AB - Congenital hypothyroidism (CH) is a neonatal disorder that is caused by a prolonged loss of thyroid hormone, which is essential for early brain development. While CH was once the leading cause of mental retardation, newborn screening for CH now allows for early identification and treatment. As a result, affected children now show normal physical and psychological development. Nevertheless, because they still undergo a brief but circumscribed period of thyroid hormone insufficiency, they are at risk for subtle selective impairments. This paper examines several of the persisting deficits observed in children with CH that was identified early in life by newborn screening as well as the relevant disease- and treatment-related factors contributing to such deficits. Highlighted will be (a) a weakness in visuospatial processing, which is associated with prenatal thyroid hormone insufficiency, (b) selective memory deficits associated with postnatal thyroid hormone insufficiencies, (c) a weakness in sensorimotor abilities also reflecting postnatal thyroid hormone insufficiencies, and (d) attention deficits, which are due to abnormal thyroid hormone levels at time of testing. Because these four disabilities implicate different neural substrates, the findings described presently will provide insights as to the specific time windows when different brain structures in the human critically need thyroid hormone. PMID- 12759832 TI - Sources of variability in sequelae of very low birth weight. AB - Few investigations have examined the specificity of sequelae of very low birth weight (VLBW, <1500 g) or sources of variability in outcome. To better understand the nature and determinants of outcome, we assessed neuropsychological and achievement skills at mean age 11 years in 62 children with <750 g birth weight, 54 with 750-1499 g birth weight, and 66 term-born controls. Distinct cognitive constructs were identified by factor analysis, and the three birthweight groups were compared on these constructs and on composite measures of achievement. Although the group with <750 g birth weight performed less well on all tests than term-born controls, group differences in a perceptual planning factor and in mathematics remained even when IQ was controlled, and deficits were more pronounced in mathematics than in reading. Results from structural equation modeling were consistent with the hypothesis that neuropsychological skills mediated the relationship between birth weight and achievement. The findings confirm the differential deficit hypothesis, support the need to consider multiple sources of variability in VLBW outcomes, and highlight the importance of neuropsychological constructs in developing an explanatory framework. PMID- 12759833 TI - Social problem-solving skills in children with traumatic brain injury: long-term outcomes and prediction of social competence. AB - The effects of childhood traumatic brain injury (TBI) on social problem-solving were examined in 35 children with severe TBI, 40 children with moderate TBI, and 46 children with orthopedic injuries (OI). The children were recruited prospectively following injuries that occurred between 6 and 12 years of age. They were followed longitudinally, and ranged from 9 to 18 years of age at the time of the current study, which occurred on average 4 years post injury. They were administered a semi-structured interview used in previous research on social problem-solving to assess the developmental level of their responses to hypothetical dilemmas involving social conflict. Children in the severe TBI group defined the social dilemmas and generated alternative strategies to solve those dilemmas at the same developmental level as did children in the OI group. However, they articulated lower-level strategies as the best way to solve the dilemmas and used lower-level reasoning to evaluate the effectiveness of the strategies. After controlling for group membership, race, socioeconomic status, IQ, and age, children's social problem-solving, and particularly the developmental level of their preferred strategies for resolving conflicts, predicted parents ratings of children's social skills, peer relationships, aggressive behavior, and academic performance. The findings indicate that children with severe TBI demonstrate selective, long-term deficits in their social problem-solving skills that may help to account for their poor social and academic outcomes. PMID- 12759834 TI - Acquired epileptiform aphasia: a dimensional view of Landau-Kleffner syndrome and the relation to regressive autistic spectrum disorders. AB - Acquired epileptiform aphasia (AEA) is characterized by deterioration in language in childhood associated with seizures or epileptiform electroencephalographic abnormalities. Despite an extensive literature, discrepancies and contradictions surround its definition and nosological boundaries. This paper reviews current conceptions of AEA and highlights variations in the aphasic disturbance, age of onset, epileptiform EEG abnormalities, temporal course, and long-term outcome. We suggest that AEA, rather than being a discrete entity, is comprised of multiple variants that have in common the features of language regression and epileptiform changes on EEG. Viewed this way, we argue that AEA can be conceptualized on a spectrum with other epileptiform neurocognitive disorders that may share pathophysiological features. The implications of this viewpoint are discussed, with emphasis on parallels between the AEA variants and regressive autistic spectrum disorders. PMID- 12759835 TI - Complications of cataract extraction. PMID- 12759836 TI - Phacoemulsification and thermal wound injury. AB - The technique of lens nucleus phacoemulsification has revolutionized cataract surgery. However, the production of ultrasound energy is associated with heat generation that can result in damage to ocular tissue, in particular the corneoscleral wound site. Thermal damage to the corneoscleral wound site may result in difficulty with wound closure and consequent risk of wound leakage, as well as damage to the adjacent corneal stroma and endothelium, fistula formation, and the induction of high degrees of post-operative astigmatism. The loss of adequate flow of irrigation fluid around the phacoemulsification tip is the key factor in the development of phacoemulsification-induced thermal injury. Use of excessive ultrasound power and production of excessive frictional forces generated by contact of the vibrating phacoemulsification needle with the irrigation sleeve are also factors involved. In the event of a "phacoburn," a specialized "gape suture" may help minimize surgically-induced astigmatism. The degree of induced astigmatism tends to wane over time; astigmatic keratotomy is an option in the setting of high degrees of residual astigmatism. PMID- 12759837 TI - Corneal edema after cataract surgery: incidence and etiology. AB - Corneal edema from inadequate endothelial pump function is one of the most common complications of cataract surgery. Various causes for this endothelial dysfunction can be divided into four categories including (a). mechanical injury, (b). inflammation/infection, (c). chemical injury, and (d). concurrent eye disease. This review serves as a basis for the diagnosis and treatment of this complication. PMID- 12759838 TI - Descemet membrane tear after cataract surgery. AB - Descemet membrane detachment is a potentially devastating complication of cataract surgery. Small localized detachments are rarely problematic, however persistent extensive detachments can affect visual acuity. In severe cases penetrating keratoplasty may be required for restoration of vision. One case of a persistent descemets membrane tear is presented and the progress after surgical repair via suture and injection of air is described. The patient was followed for 5 months after repair with persistent haze and mild corneal edema, though vision improved to 20/25+. PMID- 12759840 TI - Sterile corneal melting and necrotizing scleritis after cataract surgery in patients with rheumatoid arthritis and collagen vascular disease. AB - The onset of post-operative corneal melting and necrotizing scleritis in patients with rheumatoid arthritis and collagen vascular disease who undergo cataract surgery can have devastating ocular and systemic consequences. Even though ocular surface factors such as sicca and surgical trauma are among the important variables that contribute to this entities, signs and symptoms of systemic disease need to be thoroughly investigated in order to prevent life-threatening complications associated with these ocular manifestations. The management of surgical induced corneal melting and necrotizing scleritis in these patients, include local therapy and in most instances, systemic immuno-modulation. Moreover, the development of corneal melting and necrotizing scleritis in an otherwise "healthy" patient after uncomplicated cataract surgery, can be the first manifestation of a serious occult systemic disease. Therefore, an aggressive approach regarding the diagnosis, workup and treatment should be initiated by the ophthalmologist in order to maximize a successful ophthalmic and medical outcome. PMID- 12759841 TI - Surgical management of glaucoma in pseudophakic patients. AB - Pseudophakic patients that require surgical management of glaucoma have many of the same options as phakic patients. Considerations when performing incisional glaucoma surgery (trabeculectomy with adjunctive antimetabolites and glaucoma drainage device implantation) and laser glaucoma surgery (laser trabeculoplasty and laser cyclophotocoagulation) in pseudophakic patients are reviewed here. The decision to operate and the choice of surgical procedure must be individualized for each patient. PMID- 12759839 TI - Intraocular lens tolerance in surgery for cataracta complicata: assessment of four implant materials. AB - PURPOSE: To evaluate various available intraocular lenses (IOL) (PMMA, acrylic, heparin coated PMMA, and silicone) in patients with cataracta complicata and evaluate IOL tolerance, post-operative inflammation, cystoid macular edema, and posterior capsular opacification(PCO) requiring Nd:YAG capsulotomy. DESIGN: randomized, double masked clinical trial. PARTICIPANTS: Patients evaluated in the Ocular Immunology and Uveitis Service of the Massachusetts Eye and Ear Infirmary and deemed to need cataract surgery by standard criteria were asked to participate in the clinical trial. Patients had to have a diagnosis of chronic uveitis, which was inactive for 3 months prior to cataract surgery. RESULTS: Acrylic lenses appeared to provide the best overall results when evaluated for post-operative inflammation, PCO rates, visual acuity, and cystoid macular edema (CME) (compared to PMMA, silicone, and heparin coated PMMA). PMID- 12759842 TI - Avoiding complications in combined phacotrabeculectomy. AB - Combined phacoemulsification and trabeculectomy offers the advantage of relieving both cataract and increased intraocular pressure in one surgical sitting. The procedure however entails greater risks than either alone. An approach to minimize these risks is described. PMID- 12759843 TI - Ptosis after cataract surgery. AB - An often over-looked complication of routine cataract surgery is post-operative ptosis. This form of ptosis is often transient, although persistent ptosis may require surgical intervention. The causes include eyelid edema and hematoma, anesthesia myotoxicity, and use of a lid speculum or bridle suture. This review demonstrates that a thorough evaluation of the patient and determination of etiology of post-operative ptosis can help direct the need and timing for intervention. PMID- 12759844 TI - Neuro-ophthalmologic complications of cataract surgery. AB - Neuro-ophthalmologic complications from cataract surgery are uncommon and include central nervous system toxicity, binocular diplopia, traumatic optic neuropathy and ischemic optic neuropathy. Retrobulbar blocks may be accidentally injected into the subarachnoid space with diffusion to the brainstem. This leads to cardiovascular, respiratory, and mental status compromise. Most patients have complete recovery with adequate support. Post-operative, binocular diplopia may occur secondary to anisometropia or previously unrecognized misalignment. Periocular injection may cause paresis or fibrosis of extraocular muscles. Anterior or posterior ischemic optic neuropathy can occur in the first 6 weeks after cataract surgery with or without periocular injection. The risk to the other eye is high with subsequent contralateral cataract extraction. Post operative vision loss associated with direct traumatic needle injury is recognized immediately. Therefore, an orbital MRI may be warranted for a patient with an optic neuropathy in the first 24 hours after cataract surgery using periocular anesthesia. If evidence of needle injury is present on neuroimaging, a trial of steroids should be considered. PMID- 12759845 TI - Antibiotic therapy in post-operative endophthalmitis. AB - Endophthalmitis following routine cataract surgery is a rare but potentially devastating complication resulting in severe vision loss. Various prophylaxis treatments have been tried. However, none have been demonstrated to completely eliminate the risk. The Endophthalmitis Vitrectomy Study (EVS) helped establish clinical guidelines in the treatment of post-operative endophthalmitis in a prospective, randomized, multicenter trial. The strength and limitation of the study will be discussed as well as the current knowledge of the pharmacokinetics of intravitreous injection of antibiotics. Additionally, the common pathologic organisms, the use of antibiotic therapy, steroids, and vitrectomy will be discussed. PMID- 12759846 TI - Management of dislocated lens material. AB - Posterior dislocation of lens fragments is a well-known complication of phacoemulsification. Retained lens material in the vitreous cavity can lead to poor visual acuity related to inflammation, corneal edema, and glaucoma. Pars plana vitrectomy is frequently necessary to remove the dislocated lens fragments. The optimal timing of vitrectomy has not been firmly established. Several large studies have failed to establish an association between timing of vitrectomy and visual outcome. However, others suggest that earlier vitrectomy may lead to improved visual outcomes and lower rates of chronic glaucoma. Excellent outcomes are possible for patients undergoing vitrectomy for retained lens fragments. Retinal detachment is the most common cause of poor visual acuity in these patients. The risk of retinal detachment is correlated with vitreous manipulation by the cataract surgeon in attempts to retrieve dislocated lens material. Therefore, if both the anterior segment and vitreoretinal surgeons rigorously avoid vitreous traction in these cases, a good visual prognosis is expected. PMID- 12759847 TI - Pseudophakic cystoid macular edema. AB - Cystoid macular edema (CME) following cataract surgery has been recognized for over 50 years as an important cause of suboptimal post-operative vision. The incidence of CME varies widely, but is likely in the range of 1-2% using modern cataract extraction techniques. The diagnosis of CME can generally be made on clinical examination with evidence of perifoveal cystic spaces and can be confirmed with use of fluorescein angiography to document the classic petaloid pattern of leakage mainly into the outer retina. Leak from perifoveal vessels is induced by inflammatory mediators and results in intraretinal fluid accumulation and corresponding decrease in retinal function. The risk factors most associated with CME; rupture of posterior capsule, vitreous loss, iris incarceration, use of iris fixated lenses, active uveitis and diabetes, may all increase the potency of these mediators and exacerbate post-operative CME. The treatment of CME remains controversial but generally starts with conservative observation in isolated angiographic cases and progresses through topical non-steroidal anti-inflammatory agents (NSAIDs), topical steroids, peri-ocular steroids, systemic steroids and surgical intervention in refractory cases. Even more controversial is the role of NSAID prophylaxis peri-operatively in preventing clinical CME. Though the data is tantalizing in the short term, there is little to support the long-term benefit of such prophylaxis with respect to visual outcomes. PMID- 12759848 TI - Cystoid macular edema in aphakia and pseudophakia after use of prostaglandin analogs. AB - Prostaglandin analogs are powerful ocular hypotensive agents that may also be associated with a breakdown of the blood-retinal barrier and cystoid macular edema. The association is evident in pseudophakic or aphakic patients. This paper presents a case of unilateral latanoprost-associated clinical cystoid macular edema in a bilaterally pseudophakic patient; the only difference between the two eyes was the presence of a capsulotomy in the affected eye. We review the literature concerning prostaglandin analog-associated cystoid macular edema, as well as the evidence for prostaglandin analog-associated breakdown of the blood retinal barrier. Although some evidence is suggestive of a connection between prostaglandin analogs and cystoid macular edema, many questions concerning the complex physiology of prostanoids remain. Until our understanding of these issues is more advanced, judicious use of prostaglandin analogs in patients at risk for cystoid macular edema would be prudent. PMID- 12759849 TI - Cataract extraction and age-related macular degeneration: associations, diagnosis and management. AB - Both cataract and age-related macular degeneration (ARMD) are common findings in our aging population. At issue is the progression of ARMD following cataract extraction. The purpose of this paper is to review the data relevant to this subject and to offer suggestions as to the management of cataract associated with macular degeneration. Published data are reviewed and an attempt is made to interpret the results. Topics discussed include the epidemiology and association between the incidence of cataract and ARMD, the role of photo-oxidative damage and inflammation in the pathogenesis of ARMD, and the possible contribution of cataract extraction to the exacerbation and acceleration of ARMD through increased photo-oxidative damage and intraocular inflammation. Recommendations for the cataract surgeon and specialist are formulated on the basis of existing literature. PMID- 12759850 TI - Macular hole following cataract extraction. AB - Macular hole formation is a rare complication of cataract extraction. Although the exact etiology is unclear, macular holes can be classified into pre-existing holes that may not have been visible prior to cataract surgery, early stage macular holes that progressed to a more advanced stage, symptomatic holes following cataract extraction, and de novo symptomatic macular holes. Antero posterior (A-P) tractional forces as well as macular edema are thought to play a role in the pathogenesis of these macular holes. In the traction hypothesis, A-P forces are thought to induce either an acute detachment of the posterior cortical gel or significant traction of the vitreous gel around the fovea, resulting in formation, or rapid progression of, macular holes. In late reopening of macular holes after cataract extraction, subclinical macular edema and epiretinal membrane formation have been suggested as possible causes. Current treatment options, including combining cataract extraction with macular hole repair, are reviewed. PMID- 12759851 TI - Interventions in pseudophakic rhegmatogenous retinal detachment. AB - With refinement of vitrectomy techniques over the past 30 years, the treatment of rhegmatogenous retinal detachment (RRD) has transitioned from the almost exclusive use of extraocular techniques to the present time in which intraocular interventions are available and, in some practices, enjoy almost exclusive use for primary repair of RRD. Except for those situations in which a retinal detachment is associated with obvious immediate complications related to the cataract surgery, the considerations for the treatment of pseudophakic RRD are quite similar to those of phakic RRD. The current options are: use of an extraocular buckling device, vitrectomy-related procedures, intraocular gas injection with associated retinopexy, and combinations of these. Disagreement on the ideal intervention for pseudophakic retinal detachment is not uncommon and particularly so in those RRDs which lie somewhere in the spectrum between complex and simple. As is so often the case in the surgical treatment of disease, there may not be one ideal technique for a particular type of RRD; but knowledge of alternatives logically increases the likelihood of a successful result. The goal of this paper is to outline the clinical factors shown to be of importance in deciding which technique will have the greatest chance at not only achieving retinal re-attachment but also in optimizing long-term vision. PMID- 12759852 TI - Biostatistical analysis of pseudophakic and aphakic retinal detachments. AB - Removal of the crystalline lens increases the risk of rhegmatogenous retinal detachment (RD) by creating changes in the ocular environment that predispose to development of retinal breaks. The evolution of cataract surgery from intracapsular cataract extraction (ICCE) to extracapsular cataract extraction (ECCE) and phacoemulsification has reduced the incidence of RD, while advances in vitreoretinal surgery have resulted in improved outcomes when retinal detachment does occur. The incidence of RD varies between 0.4-3.6% for ICCE and between 0.55 1.65% for ECCE. In eyes having undergone phacoemulsification the incidence is similar to those of ECCE and ranges between 0.75-1.65%. In this article the authors review the incidence and risk factors associated with pseudophakic and aphakic RD. The risk factors discussed include pre-operative risk factors such as age, status of the fellow eye and myopia, and surgical risk factors such as vitreous loss, posterior capsular integrity and Nd : YAG capsulotomy. PMID- 12759855 TI - Corneal wound healing after excimer laser keratectomy. AB - Excimer laser keratectomy is widely used to correct refractive errors. Several complications of excimer laser keratectomy are reported including corneal infection, regression, corneal haze formation, glare and halo. Most of the complications are closely related to the corneal stromal wound healing process. In order to perform the excimer laser keratectomy with minimum complications, we should understand the mechanism of the corneal stroma wound healing process. In addition, such knowledge will help us to regulate the corneal stromal wound healing process in the future. In the present article, we discuss the molecular mechanism of the corneal stromal wound healing process after excimer laser keratectomy and its regulation by anti-inflammatory agents. PMID- 12759854 TI - LASIK vs LASEK vs PRK: advantages and indications. AB - The advent of the excimer laser as an instrument for use in reshaping the corneal stroma was a great step forward in refractive surgery. Laser energy can be delivered on the stromal surface in the photorefractive keratectomy (PRK) procedure or deeper on the corneal stroma by the means of a lamellar surgery in which a flap is created with the microkeratome in the laser in situ keratomileusis (LASIK) procedure. LASIK is currently the dominant procedure in refractive surgery. The main advantage of LASIK over PRK is related to maintaining the central corneal epithelium. This increases comfort during the early post-operative period, allows for rapid visual recovery, and reduces the wound healing response. Reduced wound healing correlates with less regression for high corrections and a lower rate of complications such as significant stromal opacity (haze). PRK, however, remains as an excellent option for mild to moderate corrections, particularly for cases associated with thin corneas, recurrent erosions, or a predisposition for trauma (Martial arts, military, etc.). Recently, a modification of PRK, laser subepithelial keratomileusis (LASEK), was introduced. In the LASEK procedure, an epithelial flap is created and replaced after the ablation. The benefits, if any, of the creation of an epithelial flap compared to traditional PRK are not fully appreciated. Advocates of LASEK suggest that there is less discomfort in the early postoperative period, faster visual recovery, and less haze compared to standard PRK for correction of similar levels of refractive error. Additional long-term clinical studies, along with laboratory research, will be crucial to validate these potential advantages of LASEK procedure. PMID- 12759856 TI - Corneal forward shift after excimer laser keratorefractive surgery. AB - The excimer laser keratorefractive surgery inevitably compromises structural integrity of the cornea by the surgical tissue subtraction and loss of integrity of Bowman's membrane. Forward shift of the cornea is commonly seen after both photorefractive keratectomy (PRK) and laser in situ keratomileusis (LASIK). Antero-posterior movement of the cornea is evaluated by measuring the posterior corneal elevation with the scanning-slit corneal topography (Orbscan). Eyes with thinner cornea, higher intraocular pressure, and higher myopia requiring greater laser ablation were more predisposed to forward shift of the cornea. After PRK, there was a trend toward progressive forward shift of the cornea, but the progression stabilized 6 months after surgery. Because progressive thinning and expansion of the cornea were not observed during the one-year observation period after PRK, the forward shift of the cornea does not represent true ectasia. Forward shift of both corneal surfaces would add to the tendency toward myopic regression after excimer laser surgery. PMID- 12759857 TI - Fourier analysis of corneal astigmatic changes following photorefractive keratectomy. AB - The purpose of this study was to evaluate the corneal irregular astigmatism following photorefractive keratectomy (PRK) for myopia. The corneal topography of 30 eyes of 26 patients was measured with the TMS-1 videokeratoscope before and 1 month after PRK. Axial dioptric data were decomposed into four components; A0 (Sphericity), C1 x 2 (Asymmetry), C2 x 2 (Regular astigmatism), and C3 (higher order irregularity) for the central 3 and 6 mm zone by Fourier series harmonic analysis. Post-operative topographies were divided into those with an irregular and those with a homogeneous pattern, and the Fourier components were compared. In the 6 mm zone, A0 was significantly decreased (P < 0.001), and C1 x 2, C2 x 2, and C3 were significantly increased (P = 0.001, 0.005, 0.002, respectively). In the 3 mm zone, A0 decreased (P < 0.001) and C1 x 2 increased (P < 0.001) significantly. C1 x 2 was correlated with the post-operative corrected visual acuity (P < 0.001, r = 0.647). The irregular pattern group had a larger C1 x 2 component (P < 0.001). The treatment displacement was not correlated with any component. In conclusion, irregular topography due to intraoperative drift or asymmetrical wound healing may play a more important role in the post-operative corneal optical property than mild treatment displacement. PMID- 12759858 TI - Comparison of ocular higher-order aberrations and visual performance between photorefractive keratectomy and laser in situ keratomileusis for myopia. AB - The purpose was to compare the ocular higher-order aberrations and the visual performance between photorefractive keratectomy (PRK) and laser in situ keratomileusis (LASIK). Ocular aberrations and visual performance were measured after PRK in 26 eyes, after LASIK in 39 eyes, and in 29 normal myopic control eyes. Ocular aberrations were measured with a Hartmann-Shack aberrometer. Visual performance was evaluated with grating contrast sensitivity, high and low contrast visual acuity, and letter contrast sensitivity under full correction with spectacles. The results were that the root mean square (RMS) values of ocular higher-order aberrations after PRK or LASIK were significantly greater than that of normal controls for both 4-mm and 6-mm zones (PRK; 0.22 +/- 0.09 and 0.85 +/- 0.24 microm, LASIK; 0.20 +/- 0.06 and 0.82 +/- 0.24 microm, normal; 0.10 +/- 0.03 and 0.33 +/- 0.11 microm. P < 0.05 between PRK and normal, LASIK and normal, One Way ANOVA on Ranks). There were no significant differences between PRK and LASIK. The ocular higher-order aberrations increased in proportion with the attempted refractive correction by PRK and LASIK. The ocular higher-order aberrations correlated better with grating contrast sensitivity, low contrast visual acuity, and letter contrast sensitivity than with high contrast visual acuity. There was no difference among normal, PRK and LASIK in all the visual function tests, except between normal and PRK, or between normal and LASIK with letter contrast sensitivity. In conclusion, there was no difference in both ocular higher-order aberrations and visual performance between PRK and LASIK. The result suggests that surgeons can choose refractive procedures according to the corneal conditions or daily activities of patients. PMID- 12759859 TI - Measuring of ocular wavefront aberration in large pupils using OPD-scan. AB - Wavefront analysis using a scanning slit refractometer (the OPD-Scan) was evaluated in both artificial and human eyes, in comparison with the Hartmann Shack wavefront sensor (H-S) currently often used. Comparison of different methods and configurations carried out in the artificial eyes yielded basically the same results in moderate refractive error cases. There was a closer match between the RMS (root mean square) wavefront error obtained by OPD-Scan and H-S in the 6 mm pupils than in the 4 mm pupils of the same normal human eyes. Although OPD-Scan employs a different approach in determining aberration, the aberration value is similar to that of H-S, especially in large pupils. PMID- 12759860 TI - Higher-order wavefront aberrations induced by small ablation area and sub clinical decentration in simulated corneal refractive surgery using a perturbed schematic eye model. AB - The purpose was to investigate the effect of decentered ablation in myopic corneal refractive surgery by simulation. Wavefront aberrations in a small pupil area (radius: 2 mm) for photopic vision and in a large pupil area (radius: 3 mm) for mesopic vision with simulated refractive surgery were analyzed using Zernike polynomials. Radii of ablation were 3, 2.5 and 2 mm. Decentrations of ablation were 0 mm and 0.5 mm. Change of the surface shape by ablation was considered a perturbation, while Gullstrand's schematic eye was used as un-perturbed optics. For photopic vision, wavefront aberrations were about the same as with un perturbed optics. For mesopic vision, the results were heavily dependent on the radius of ablation area. When the radius was 3 mm, wavefront aberrations did not increase very much compared to un-perturbed optics. When the radius was smaller than 3 mm, spherical aberration was induced by centered ablations, and coma was also induced by decentered ablations. In conclusion, small ablation areas or subclinical decentrations of ablations could cause serious amounts of wavefront aberrations to the optics of the eye in the simulations. PMID- 12759861 TI - The challenge of severe dry eyes--not for the fainthearted. PMID- 12759863 TI - Metastatic tumors of the orbit: a retrospective study. AB - PURPOSE: To present the clinical features and facts on prognosis, therapy and diagnostics of orbital metastases treated in the Orbital Center Kiel. METHODS: We reviewed the records of 20 patients with orbital metastases (ca. 2% of all orbital patients seen in a 10-year period). RESULTS: The mean age at first manifestation of orbital symptoms was 64 years. On average, 64 months was the period of time between the onset of the primary disease and orbital manifestation. The sites of the primary tumors were breast (8), choroid (2), prostate (2), gastrointestinal tract (3), lung (2), urinary bladder (1), Schmincke tumor of the throat (1) and unknown origin (1). Eleven patients were female, 9 male. The main primary symptoms consisted of lid swelling (11), red eye (9), diplopia (10), and proptosis (7). Although the majority (16/20) were treated by chemotherapy, radiotherapy or surgery, 19 patients died after a mean survival time of 14.7 months. CONCLUSION: Orbital metastases remain an unfavorable prognostic factor, but the diagnosis and treatment are still important in order to prevent loss of vision and improve the patient's quality of life. As previous studies have shown, the overall survival rate is still as limited as nearly half a century ago. PMID- 12759864 TI - Young men with epiphora: management by an innovative surgical technique (the Safdarjang suture). AB - A congenital medial canthal anomaly was identified as the cause of epiphora in 18 young adult male patients. This anomaly consists of anomalous anterior displacement of the medial palpebral ligament causing a lack of adequate apposition of the medial end of the lower lid to the globe. The lacrimal punctum does not dip into the lacrimal lake, thereby causing epiphora. An innovative surgical technique is described involving the passage of a suture in the pretarsal plane along the lid margin to correct the deformity. PMID- 12759862 TI - A randomized study comparing DCR with and without excision of the posterior mucosal flap. AB - AIM: A prospective randomized study was carried out to compare the success rate of external dacryocystorhinostomy (DCR) with and without excision of the posterior sac mucosal flap. METHODS: Forty patients (Group A) underwent DCR without excision of the posterior sac mucosal flap, and the results obtained were compared with those of another series of 40 patients (Group B) where DCR was performed with excision of the posterior sac flap. A large posterior flap was excised from the lacrimal sac and a large, mobile, anteriorly-hinged, nasal mucosal flap was fashioned. The latter was joined to the anterior sac flap using a 6/0 double-armed polyglycolic acid suture. RESULTS: Four patients in Group B had signs and symptoms of tearing (90% success rate) at the end of the follow-up period (mean 11.05 months, range 9-14 months). Group A had six patients with recurrent signs and symptoms (85% success rate) by the end of a mean follow-up period of 11.3 months (range 9-14 months). CONCLUSION: Excision of the posterior sac mucosa may improve the success rate of external DCR. PMID- 12759865 TI - Subcutaneous abdominal artificial tears pump-reservoir for severe dry eye. AB - PURPOSE: To assay a totally implanted pump-reservoir unit placed under the subcutaneous tissue of the abdomen for providing artificial tears to the ocular surface in patients with severe dry eye. DESIGN: Prospective non-randomized comparative (self-controlled) trial. PARTICIPANTS: Six patients with severe dry eye, in whom intermittent moistening of the ocular surface with current collyria was clinically unsatisfactory. METHODS: The six patients were treated by implanting an artificial tear pump-reservoir unit under the subcutaneous tissues of the abdomen. The reservoir is operated by a gas pump, which pumps artificial tears to the eye from a 60-ml reservoir through a silicone tube leading subcutaneously from the reservoir, via the chest, neck and lateral part of the head, and entering the conjunctival sac over the lateral canthal ligament. The catheter is anchored to the aponeurosis of the temporal muscle at the lateral rim of the orbit with a butterfly sleeve. The terminal portion of the tube runs freely along the upper conjunctival fornix, and pours 1.5 ml/day of the artificial tears into the ocular lacrimal basin with a constant flow rate. The reservoir must be refilled by percutaneous injection of artificial tears every 45 days. MAIN OUTCOME MEASURES: Schirmer test, corneal fluorescein staining, lacrimal film breakup time, lacrimal osmolarity, corneal impression cytology, best-corrected visual acuity, dryness sensation and blepharospasm before and after lacrimal reservoir implantation. RESULTS: The lacrimal subcutaneous abdominal reservoir was well tolerated with little discomfort. A delivery of 1.5 ml/day was enough to maintain a comfortable wet eye. After an average follow-up of 15 months the signs and symptoms of dry eye were dramatically improved. Four of the patients had a severe blepharospasm, which disappeared some weeks after the implantation of the lacrimal abdominal reservoir. CONCLUSION: These are the first totally implanted lacrimal reservoirs in human beings. They have proved to be a good solution for severe dry eye. At present, this method is the only one that permits a maintained wet eye surface, and the performance of corneal, conjunctival, limbal and amniotic membrane transplants in total or almost total xerophthalmia. It may also be a good solution for some of the so-called essential blepharospasms, which are frequently triggered by an underlying dry eye. PMID- 12759866 TI - Dirofilarial infection of the orbit. AB - PURPOSE: We report on two cases of dirofilaria involving the orbit. There have been no previous case reports of superior rectus involvement in the literature. METHODS: The presenting history, ocular examination, relevant investigations and treatments are discussed. RESULTS: In both cases the lesion was debulked surgically with subsequent histopathological identification of a dirofilarial worm. Postoperatively both patients were relieved of their presenting symptoms with no recurrence of the lesion. CONCLUSIONS: Dirofilarial involvement of the orbit is uncommon. It should be considered as a rare infectious form of specific orbital inflammation and considered in the differential diagnosis of orbital inflammatory lesions. PMID- 12759867 TI - Conjunctival malignant melanoma. AB - A 65-year-old woman was diagnosed clinically to have a tarsal conjunctival malignant melanoma of the upper eyelid. She also had multiple, diffusely pigmented, flat conjunctival lesions. No local or distant metastasis was detected. She was treated conservatively by a combination of surgery and cryotherapy as this was her only seeing eye. Histopathology of the tumor tissue and biopsy specimens of the flat conjunctival lesions proved them to be conjunctival malignant melanoma and primary atypical melanosis with atypia, respectively. Over a follow-up of two years, no clinical recurrence of the tumor was seen. To conclude, a malignant melanoma arising from the tarsal conjunctiva is extremely rare. The goal of treatment should be eradication of the tumor as well as preservation of a functionally and cosmetically acceptable eye. PMID- 12759868 TI - Complications of the orbital floor and maxillary sinus 30 years after Coe-Pak misplacement in the management of pan-facial fractures. AB - We present the case of a woman who had sustained pan-facial fractures in a road traffic accident 30 years previously, and describe the ensuing unusual problems with the orbital floor and maxillary sinus as a consequence of unrecognised misplacement of a dental periodontal dressing material into the sinus. The subsequent management is discussed. PMID- 12759869 TI - Vacuum Assisted Closure (VAC) dressing for skin graft application following exenteration of the orbit. PMID- 12759870 TI - Isolated abscess of extraocular muscle in two young boys: clinical and imaging features. AB - PURPOSE: To report two cases of a cystic isolated extraocular muscle abscess in a young boy. METHOD: Case report. RESULTS: Two cases of young boys with a cystic isolated extraocular muscle abscess are described. Each presented as an inflammatory lesion in the orbit that had been stable for approximately three weeks. Neurodiagnostic imaging of the two cases revealed a cystic lesion in close proximity to an extraocular muscle. An ultrasound-guided aspiration of each lesion revealed material that, when cultured, grew Staphylococcus aureus. Both patients recovered with appropriate antibiotic therapy. No other systemic illness could be identified in either case. CONCLUSION: These cases illustrate that an isolated abscess involving an extraocular muscle may present like an inflammatory cyst even without any evidence of systemic illness. When evaluating and managing such lesions, the authors recommend the use of ultrasound-guided aspiration to avoid complications arising from delay in diagnosis and inappropriate treatment. PMID- 12759871 TI - [Lack of physicians in anesthesia and its reasons]. PMID- 12759872 TI - [Shortage of anaesthesiologists in Germany]. AB - After the glut of doctors in the past, we find a shortage of doctors in the present time. Approximately only 60% of all doctors would decide to become a doctor again. These alarming news should urge us to improve the situation for the rising generation of doctors especially for young anaesthesiologists. The actual situation in the hospital is rather unattractive and there is increasing risk that we will not have a sufficient number of anaesthesiologists in our hospitals in the near future. Improvement of the financial situation is without doubt important--this must include payment according to performance principle rather than payment with regard to age. Moreover, training programs (e. g. financial support of meetings) of the young anaesthesiologists, working conditions for all staff members (e. g. limitation of overtime), and importance of the role of anaesthesia must be modified to improve motivation and to guarantee high standards of quality of anaesthesia for the future. PMID- 12759873 TI - [Guidelines for the prevention of catheter-related infections]. AB - The revised American guidelines of the Centers for Disease Control and Prevention (CDC) 'Guidelines for the prevention of catheter-related infections' were published on August 9, 2002. The goal of the guidelines is to promote patient safety and to decrease preventable infections by providing evidence-based recommendations for those healthcare providers who insert and maintain intravascular catheters. Since the guidelines were last issued in 1996, the science of prevention has yielded several important advances in new technology to prevent infection. In addition this update makes allowances for the fact that many new randomised controlled trials for the prevention of catheter-related infection were published in the last six years. Furthermore, several approaches, long known to be effective, need more emphasis by practitioners. The following publication summarizes the state of knowledge about the most important controversial prevention strategies and explains the recommendations. PMID- 12759874 TI - Pharyngeal mucosal pressures with the laryngeal tube airway versus ProSeal laryngeal mask airway. AB - We tested the hypothesis that mucosal pressures are higher for the laryngeal tube airway trade mark than the ProSeal laryngeal mask airway. Fifteen fresh cadavers were studied. Microchip pressure sensors were attached to the laryngeal tube airway and ProSeal laryngeal mask airway at four similar anatomical locations (base of tongue, lateral pharynx, posterior pharynx and posterior hypopharynx) and three dissimilar locations (laryngeal tube airway trade mark, anterior and lateral hypopharynx; ProSeal laryngeal mask airway, pyriform fossa). The cuff volume (laryngeal tube airway, < or = 140 ml; ProSeal laryngeal mask airway, < or = 30 ml) was adjusted until the oropharyngeal leak pressure was 15 cm H2O and the mucosal pressures recorded. This was repeated at an oropharyngeal leak pressure of 20, 25 and 30 cm H2O. Mucosal pressures in the lateral pharynx were always similar. Mucosal pressures at the base of tongue and posterior pharynx were similar for the laryngeal tube airway and ProSeal laryngeal mask airway at 15 and 20 cm H2O, but were higher for the laryngeal tube airway at 25 cm H2O at 30 cm H2O. Mucosal pressures in the posterior hypopharynx were always higher for the laryngeal tube airway (all: p < 0.03). Mean mucosal pressures for the laryngeal tube airway ranged from 8-31, 2-13 and 15-41 cm H2O for the base of tongue, lateral pharynx and posterior pharynx (proximal cuff) respectively and 3-7, 3-7 and 7-18 cm H2O for the anterior, lateral and posterior hypopharynx (distal cuff) respectively. Mean mucosal pressures for the ProSeal laryngeal mask airway ranged from 6-23, 3-10, 8-25, 6-17 and 2-8 cm H2O for the base of tongue, lateral pharynx, posterior pharynx, pyriform fossa and posterior hypopharynx respectively. We conclude that mucosal pressures are higher for the laryngeal tube airway, particularly when oropharyngeal leak pressure greater than 25 cm H2O. This suggests that mucosal ischemic injury will be more common with the LTA than the PLMA. PMID- 12759875 TI - [Correlation of a computerized SOFA score and derived measures with length of stay at an operative ICU]. AB - OBJECTIVE: The purpose of this study was to assess whether or not the information on progression of illness and therapeutic interventions included in a computerized SOFA (Sequential Organ Failure Assessment) score and derived measures (maximum SOFA, total maximum SOFA and delta SOFA) influences the correlation with length of stay (LOS) at an operative intensive care unit (ICU). METHODS: During a period of one year 524 patients aging over 18 years who stayed more than 24 hours at an operative ICU were included into this study. Based on SQL scripts (Structured Query Language), a computerized SOFA score and the derived measures "maximum-SOFA", "total-maximum-SOFA" and "delta-SOFA" of all patients were retrospectively calculated for each day at ICU using routine data recorded with the patient data management system ICUData (IMESO GmbH, Huttenberg, Germany). "Maximum-SOFA" and "total-maximum-SOFA" are measures for the severity of illness or cumulative organ dysfunction. "Delta-SOFA" measures the magnitude of organ dysfunction developing during ICU stay and is therefore potentially influenced by therapeutic measures. Analysis was based on exact LOS in hours. RESULTS: Mean LOS of the whole collective was 5.2 +/- 6.8 days (median 2.8, range 1 - 51). Correlation of LOS depending on score results increased from "admission SOFA" (r = 0.280), to "maximum-SOFA" (r = 0.444), "total-maximum-SOFA" (r = 0.503), and "delta-SOFA" (r = 0.576). CONCLUSION: Therefore, the sum of information on progression of illness and therapeutic interventions included in derived measures improves the correlation with LOS at an ICU compared to the "admission-SOFA". PMID- 12759876 TI - [Tumor pain therapy]. PMID- 12759878 TI - Predicting cardiovascular risk factors from plasma cortisol measured during oral glucose tolerance tests. AB - Increasing evidence suggests that activation of the hypothalamic-pituitary adrenal (HPA) axis may contribute to the pathogenesis of the metabolic syndrome and obesity. The mechanisms are unknown but may involve alterations in the metabolic responses to feeding that interact with the HPA axis. As it is known that plasma cortisol falls during an oral glucose tolerance test (OGTT), changes in cortisol measured during an OGTT may be altered in the metabolic syndrome. We measured changes in plasma cortisol during OGTTs in a large study of 593 men and women to determine correlates of changes in cortisol with features of the metabolic syndrome and the extent to which these relationships are confounded by obesity. In men and women, higher cortisol area under the curve (AUC) during the OGTT was associated with higher glucose AUC and higher systolic blood pressure. Higher cortisol AUC was associated with reduced insulin increment in men, but higher 2-hour insulin and insulin AUC in women. However, the decline in plasma cortisol after glucose administration was poorly predictive of features of the metabolic syndrome. Obesity was associated with lower cortisol AUC but not with percentage decline in cortisol. Plasma cortisol and obesity had independent effects on plasma glucose and were the strongest predictors of plasma glucose in multiple regression analysis. Measurements of plasma cortisol during the OGTT reinforce the previously observed relationships of activation of the HPA axis in the metabolic syndrome. However, the altered HPA response to feeding does not appear to be primarily responsible for HPA activation in subjects with the metabolic syndrome. PMID- 12759877 TI - Vitamin D receptor and aromatase gene interaction and bone mass in older African American women. AB - Aromatization of androgens by the CYP19 gene product, aromatase, is the major source of endogenous estrogen in postmenopausal women. We determined whether an Arg(264)Cys polymorphism in the CYP19 gene is associated with bone mineral density (BMD) and bone loss in older women. Because vitamin D regulates CYP19 gene expression, we also tested for an interaction with a translation start site polymorphism in the vitamin D receptor (VDR) gene. Hip BMD was measured twice, an average of 1.9 years apart, in 100 African-American women aged > or =65 years. Neither polymorphism alone was significantly associated with BMD or bone loss. BMD measurements in women with the less frequent allele at both loci were 0.5 to 1.3 SD lower than in women with neither or only a single rare allele (P <.001 for interaction). These women also experienced more rapid hip bone loss than other women (P <.05 for interaction). We conclude that VDR and CYP19 gene polymorphisms may jointly influence bone mass and the rate of bone loss in older African American women. PMID- 12759879 TI - A 48-hour exposure of pancreatic islets to calpain inhibitors impairs mitochondrial fuel metabolism and the exocytosis of insulin. AB - Genetic variation in the gene for a cytosolic cysteine protease, calpain-10, increases the susceptibility to type 2 diabetes apparently by altering levels of gene expression. In view of the importance of altered beta-cell function in the pathophysiology of type 2 diabetes, the present study was undertaken to define the effects on insulin secretion of exposing pancreatic islets to calpain inhibitors for 48 hours. Exposure of mouse islets to calpain inhibitors (ALLN, ALLM, E-64-d, MDL 18270, and PD147631) of different structure and mechanism of action for 48 hours reversibly suppresses glucose-induced insulin secretion by 40% to 80%. Exposure of islets to inhibitors of other proteases, ie, cathepsin B and proteasome, did not affect insulin secretion. The 48-hour incubation with calpain inhibitors also attenuates insulin secretory responses to the mitochondrial fuel alpha-ketoisocaproate (KIC). The same incubation also suppresses glucose metabolism and intracellular calcium ([Ca(2+)](i)) responses to glucose or KIC in islets. In summary, long-term inhibition of islet calpain activity attenuates insulin secretion possibly by limiting the rate of glucose metabolism. A reduction of calpain activity in islet could contribute to the development of beta-cell failure in type 2 diabetes thereby providing a link between genetic susceptibility to diabetes and the pathophysiologic manifestations of the disease. PMID- 12759880 TI - Fat feeding impairs glycogen synthase activity in mice without effects on its gene expression. AB - To examine whether the effects of high-fat feeding on glycogen synthase (GS) activity and mRNA levels differ between diabetes-prone (C57BL/6J) and diabetes resistant mice (NMRI), we measured GS activity and mRNA levels in muscle from C57BL/6J and NMRI mice fed a high-fat or normal chow diet for 3, 6, and 15 months. As compared with chow feeding, fat feeding increased plasma insulin levels in C57BL/6J mice at 15 months (464 +/- 29 v 267 +/- 47 pmol/L, P =.005), which was associated with elevated plasma glucose levels at 15 months (5.3 +/- 0.3 v 3.8 +/- 0.2 mmol/L, P =.001). Fat feeding increased plasma insulin levels also in NMRI mice at 15 months (705 +/- 145 v 275 +/- 64 pmol/L, P =.01) without, however, a rise of plasma glucose levels. In parallel with increased insulin levels, decreased muscle GS fractional velocity (FV) was observed at 6 (49.0% +/- 2.6% v 69.1% +/- 7.3%, P =.04) and 15 (45.8% +/- 1.8% v 53.4% +/- 1.6 %, P <.01) months but not at 3 months in the fat-fed C57BL/6J mice. Similarly, there was a significant decrease in GS fractional activity at 3 (57.9% +/- 4.3% v 70.4% +/- 2.6 %, P <.03) and 15 (47.3% +/- 2.4% v 56.4% +/- 2.1%, P =.02) but not at 6 months in the fat-fed NMRI mice. The decrease in GS activity was not associated with changes in mRNA levels at any time points. We conclude that (1) fat feeding results in similar elevation of plasma insulin levels and impairs GS activity in C57BL/6J and NMRI mice, and (2) the changes in GS activity do not involve effects on gene expression. PMID- 12759882 TI - High glucose concentration decreases insulin-like growth factor type 1-mediated mitogen-activated protein kinase activation in bovine retinal endothelial cells. AB - Clinical trials have incontrovertibly demonstrated that the onset and progression of diabetic retinopathy (DR) is influenced by the control of glucose levels in patients. In the present study, we examined the effect of glucose concentration on the responsiveness of bovine retinal endothelial cells (BREC) to insulin-like growth factor type 1 (IGF-1). Retinal endothelial cells were isolated from bovine retina and cultured in 5 or 20 mmol/L glucose with or without 100 ng/mL IGF-1. The level of cell growth and p42/44 and p38 mitogen-activated protein kinase (MAPK) activation was determined using the alamarBlue (Serotech) assay and Western blotting, respectively. IGF-1 significantly enhanced cell growth in BREC exposed to 5 mmol/L glucose but not in cells exposed to high glucose concentrations (20 mmol/L). IGF-1 induced a transient activation of p42/44 MAPK, with peak activation at 15 minutes in cells exposed to 5 mmol/L glucose; however, no increase in p42/44 MAPK was evident at the higher glucose concentration of 20 mmol/L. There was no significant change in the level of p38 MAPK during the time period examined when IGF-1 was also present. However, high glucose concentrations alone increased the level of p38 MAPK after 60 minutes and the level of p42/44 MAPK after only 15 minutes exposure in 20 mmol/L glucose. Thus, BREC exposed to high glucose concentrations are not sensitive to IGF-1 and this is due, at least in part, to a reduced activation of the p42/44 MAPK pathway. Furthermore, the presence of IGF-1 appears to exert a protective effect on the cells in high glucose concentration by preventing progression through the cell cycle. PMID- 12759881 TI - Regulation of skeletal muscle morphology in type 2 diabetic subjects by troglitazone and metformin: relationship to glucose disposal. AB - The goal of this work was to compare the effects of different antidiabetic therapies on the phenotype of skeletal muscle in type 2 diabetic subjects failing sulfonylurea therapy. Subjects were treated with a thiazolidinedione (troglitazone, TGZ) or a biguanide (metformin, MET) in addition to glyburide for 3 to 4 months. Insulin action was determined with a hyperinsulinemic (300 mU. m( 2). min(-1)) euglycemic (5.0 to 5.5 mmol/L) clamp. Biopsies were obtained from the vastus lateralis muscle for morphological analysis. Despite similar glycemic control, relative increases in the insulin-stimulated glucose disposal rate (GDR) were greater after TGZ treatment (37 +/- 8% increase, P <.05) than after MET (21 +/- 11%, P <.05). Neither treatment had any effect on fiber type composition of the muscle. Capillary density was reduced in diabetic subjects compared to a nondiabetic group (P <.01) and was increased with TGZ treatment (P <.05), while MET was without significant effect. Diabetic muscle also displayed a lower mitochondrial volume density that was unaltered by either treatment. Both TGZ and MET therapy resulted in a reduction in the lipid content of muscle (percent fiber volume as lipid droplets); the relative decrease tended to be greater for TGZ ( 33% v -23% for MET). The relative (%) improvement in GDR was correlated with the change in lipid content (r = -0.756, P <.05) after TGZ treatment; no such relationship was observed for MET. From these results we conclude that the higher potency of TGZ to increase capillary density and reduce the lipid content of muscle may contribute to its greater ability to improve glucose disposal in skeletal muscle of type 2 diabetic individuals. PMID- 12759883 TI - Body composition and energy expenditure: relationship and changes in obese subjects before and after biliopancreatic diversion. AB - Changes in total and segmental body composition were studied in 101 obese women before and 2, 6, 12, and 24 months after biliopancreatic diversion (BPD) and data 24 months after surgery were matched to 53 control subjects. The patients were studied by anthropometry, indirect calorimetry, and double-emission x-ray absorptiometry (DXA). The combination of calorimetry and body composition analysis allowed estimation of visceral and muscle lean mass. We observed a significant (analysis of variance [ANOVA]: P <.05) progressive reduction of fat and lean body mass (LBM) following BPD, with stabilization of both parameters between 12 and 24 months at levels not different from controls. Fat loss was significant in the arms, legs, and trunk segments. After 24 months, there was no significant difference in segmental fat mass between post-BPD patients and controls. Calorimetric data seem to confirm lean body mass (LBM) reduction. Visceral lean mass (kg) was significantly reduced from 8.1 +/- 2.2 in obese subjects to 6.5 +/- 1.8 in post-BPD patients at 24 months (P <.05); the control value was 7.2 +/- 1.8. Muscular lean mass (kg) was also significantly reduced, from 50.2 +/- 5.8 to 39.8 +/- 5.7 in the same subjects (P <.05), with a control value of 42.5 +/- 5.9. The decrease in muscle and visceral LBM reached control values without significant differences. Viscera/muscle ratio in pre-BPD patients was preserved in post-BPD patients at 24 months, but it was reduced during weight loss. Body composition studies showed a logarithmic relationship between fat and lean mass and a physiological contribution of lean mass to weight loss in the BPD patients. In conclusion, weight loss after BPD was achieved with an appropriate decline of LBM and with all parameters reaching, at stable weight, values similar to weight-matched controls. PMID- 12759884 TI - Lipoproteins abnormalities in obese insulin-resistant dogs. AB - Many studies have shown that obesity and low insulin sensitivity are associated with lipoprotein abnormalities, which are risk factors for coronary heart disease. The effects of insulin resistance on lipoprotein metabolism were investigated in hyperenergetic-fed beagle dogs, a new model of insulin resistance. Insulin resistance was assessed by the 3-hour euglycemic hyperinsulinemic glucose clamp technique. Lipoproteins were separated by fast protein liquid chromatography (FPLC) and lipid composition of the different lipoproteins was determined by enzymatic methods. Hyperenergetic diet was associated with a 43% +/- 5% increase in dog body weight and a reduction in insulin-mediated glucose uptake (28 +/- 3 to 16 +/- 1 mg. kg(-1). min(-1), P <.05). Low insulin sensitivity associated with obesity was related to an increase in plasma triglyceride (TG) through an increase in very-low-density lipoprotein (VLDL)-TG (0.071 +/- 0.020 v 0.382 +/- 0.242 mmol/L, P <.05) and high-density lipoprotein (HDL)-TG (0.025 +/- 0.012 v 0.242 +/- 0.143 mmol/L, P <.05). Other lipid abnormalities common in insulin resistant humans were also found: lower plasma HDL-cholesterol (4.690 +/- 0.151 v 3.937 +/- 0.141 mmol/L, P <.05) and higher plasma nonesterified fatty acids (NEFA) (0.974 +/- 0.094 v 1.590 +/- 0.127 mmol/L, P <.05) levels. These data show that this model of the insulin-resistant obese dog could be useful in studying insulin resistance-associated dyslipidemia. PMID- 12759885 TI - Metabolic effects of chronic glipizide gastrointestinal therapeutic system on serum glucose, insulin secretion, insulin sensitivity, and hepatic insulin extraction in glucose-tolerant, first-degree relatives of African American patients with type 2 diabetes: new insights on mechanisms of action. AB - We examined the long-term metabolic effects of a potent sulfonylurea (SU), glipizide gastrointestinal therapeutic system (glipizide GITS) in normal glucose tolerant (NGT), first-degree relatives of African American patients with type 2 diabetes in a randomized, placebo-controlled, double-blind manner for 24 months and 6 months after discontinuation of glipizide GITS. Fifty NGT African American first-degree relatives (n = 50)) were randomized to receive either glipizide GITS (GITS, 5 mg/d) or identical placebo (PLAC). The NGT consisted of NGT/GITS (n = 16; mean age, 43.1 +/- 8.7years; body mass index [BMI], 34.8 +/- 10) and NGT/PLAC (n = 34; 45.5 +/- 9.7 years; BMI, 31.3 +/- 3.1years). Each of the subjects underwent an oral glucose tolerance test (OGTT) and frequently sampled intravenous glucose tolerance test (FSIGT) at baseline and at yearly intervals for 2 years. Insulin sensitivity (Si) and glucose effectiveness (Sg) were determined by Bergman's minimal model method. Hepatic insulin extraction (HIE) was calculated as the molar ratio of C-peptide and insulin. The mean fasting serum glucose, insulin, and C-peptide levels in the NGT/GITS were not different from that of the NGT/PLAC. After oral glucose challenge, mean serum glucose responses slightly increased (P = not significant [NS]) at 12 and 24 months in the NGT/GITS group when compared with the baseline, 0 month, but remained unchanged in the NGT/PLAC group. In addition, serum insulin and C-peptide responses significantly increased in the NGT/GITS group, but were unchanged in the NGT/PLAC group at 12 and 24 months versus 0 month. The HIE, during OGTT, decreased by 30% from the baseline (0 month) values in the NGT/ GITS, but remained unchanged in the NGT/PLAC group at 12 and 24 months. Mean Si decreased by 30% from the baseline in the NGT/GITS group by 12 and 24 months, but remained unchanged in the NGT/PLAC group. However, the disposition index (DI) remained normal in the NGT/GITS and the NGT/PLAC groups. The DI data in the NGT/GITS group suggested that beta cells maintained the ability to compensate for the lower Si during the chronic GITS administration in our high risk African Americans. Chronic GITS was well tolerated without any symptoms of either hypoglycemia or weight gain in the NGT/IGTS group. After discontinuation of GITS, the altered metabolic parameters significantly improved, returning to baseline values in the NGT/IGTS group in 6 months. In summary, chronic glipizide GITS administration (5 mg/d) was associated with increased beta-cell secretion, peripheral hyperinsulinemia, reduced Si, and reduced HIE in glucose-tolerant, first-degree relatives of African American patients with type 2 diabetes. These metabolic changes were reversible within 6 months after discontinuation of glipizide GITS. Our study defines a unique mode of action of glipizide GITS in African Americans at high risk for type 2 diabetes. We conclude that the use of glipizide GITS in the primary prevention of type 2 diabetes in nondiabetic first-degree relatives of patients with type 2 diabetes impaired glucose homeostasis. PMID- 12759887 TI - Alterations of bone mineral density and bone metabolism in patients with various grades of chronic pancreatitis. AB - The aim of this study was to examine bone mineral density (BMD) and bone metabolism in patients with chronic pancreatitis to determine if increased severity of the disease would correlate with increased bone loss. Between October 1999 and September 2000, we investigated 42 patients with an average age of approximately 53 years suffering from chronic pancreatitis, as well as 20 healthy male controls with an average age of 49 years. Dual energy x-ray absorptiometry (DEXA) was performed on patients and controls, and serum levels of parathyroid hormone (PTH), osteocalcin (OC), carboxy-terminal propeptide of type I procollagen (CICP), bone-specific alkaline phosphatase (BAP), 1,25(OH)(2) vitamin D(3) and 25(OH) vitamin D(3), as well as fecal elastase 1 were also determined. The severity of chronic pancreatitis in patients was determined via endoscopic retrograde cholangiopancreatography (ERCP) and assigned to 1 of 3 grades based on the Cambridge classification. BMD of patients with chronic pancreatitis was markedly decreased compared to controls (means in patients: DEXA lumbar vertebra anterior/posterior (LV ap) 96.8% +/- 4.2%, DEXA Ward's triangle (WARD) 92.2% +/- 5.2%; controls: DEXA LV ap 98.7% +/- 3.7%, DEXA WARD 97.1% +/- 3.1%; P <.05 and P <.0001) and correlated with the various Cambridge-grades (DEXA LV ap and DEXA WARD, P <.01). Fecal elastase 1 showed sensitivities of 14%, 87%, and 95% for the Cambridge-grades I, II, and III, respectively, and correlated with this classification of severity of chronic pancreatitis (P <.01). Furthermore, fecal elastase 1 of patients correlated the same way with both D(3)-vitamins (P <.01), as well as with parameters of BMD (P <.01). If fecal elastase 1 in patients was below 200 micro g/g, then the BMD and vitamin D(3) values were also significantly decreased compared to those with fecal elastase 1 above 200 micro g/g. In patients with Cambridge grades II and III 1,25(OH)(2)D(3) was markedly decreased (26.7 +/- 7.7 pg/mL and 27.6 +/- 9.0 pg/mL) compared to those with Cambridge grade I (38.0 +/- 10.5 pg/mL; between I and II, P =.027; between I and III, P =.033). 25(OH)D(3) was not significantly different within the various Cambridge groups (P =.07). Compared to controls, both D(3) vitamins, as well as fecal elastase 1, were extremely low (means in patients: fecal elastase 1, 140.7 +/- 75.7 micro g/g; 1,25(OH)(2)D(3), 29.9 +/- 9.5 pg/mL; 25(OH)D(3), 26.7 +/- 9.7 nmol/L; controls: fecal elastase 1, 694.9 +/- 138.6 micro g/g; 1,25(OH)(2)D(3), 67.5 +/- 4.3 pg/mL; 25(OH)D(3), 69.5 +/- 13.5 nmol/L). A significant correlation was observed between increased severity of chronic pancreatitis based on both endoscopic retrograde cholangiopancreatography and levels of fecal elastase 1, with decreased circulating levels of vitmain D(3) and decreased BMD. This supports a connection between the inflammatory destruction of the pancreas (Cambridge classification), exocrine pancreatic insufficiency (fecal elastase 1), altered levels of vitamin D metabolites, and loss of skeletal mass. PMID- 12759886 TI - Determinants of insulin secretion after renal transplantation. AB - The high prevalence of post-transplant glucose intolerance and insulin resistance (IR) is associated with older age, family history of diabetes, immunosuppressive drugs, and antihypertensive therapy. However, the potential determinants of post transplant beta-cell dysfunction are largely unknown. The objective of the present study was to address this issue in detail. A total of 167 previously nondiabetic renal transplant recipients underwent a 75-g oral glucose tolerance test (OGTT)10 weeks after transplantation. Serum glucose and insulin were measured at 0, 1, and 2 hours. Three insulin release indices (Secr(AUC), Secr(1.phase), and Secr(2.phase)) were calculated to assess the insulin secretory response as the dependent variable. To account for variations in insulin sensitivity (IS), beta-cell function was also estimated as the disposition index (DI); the product of the IS index (ISI(TX)) and Secr(1.phase). Increasing age was strongly and independently associated with a blunted insulin secretory response even after adjustment for IS (P =.001). An 80-year-old recipient had an approximately 50% lower insulin release than a 20-year-old individual, based on the linear regression model. Cytomegalovirus (CMV) disease and treatment with furosemide were both independently associated with beta-cell dysfunction (DI; P <.001 and P =.008). Patients treated with angiotensin-converting enzyme (ACE) inhibitors had an enhanced absolute insulin release, but the DI was similar in both treated and untreated recipients. We conclude that older age is an important determinant of beta-cell dysfunction after renal transplantation. CMV disease and treatment with furosemide may also negatively influence pancreatic insulin release in renal transplant recipients. PMID- 12759888 TI - An intracellular modulation of free radical production could contribute to the beneficial effects of metformin towards oxidative stress. AB - Metformin (dimethylbiguanide) is an antihyperglycemic agent used in type 2 diabetes. Beyond its action on glycemic control, metformin exhibits other intrinsic effects that could play a role in prevention against diabetes complications. Some studies thus reported an improvement in the antioxidant status in patients treated with metformin. This might be in part related to its property to limit formation of advanced glycation end products (AGEs) and to decrease the overproduction of free radicals in diabetic subjects. The aim of this study was to investigate the in vitro ability of metformin to modulate the action of reactive oxygen species (ROS) generated either by water gamma radiolysis or by stimulated human leukocytes. Our results showed that metformin at pharmacologically relevant concentrations was in vitro able to scavenge hydroxyl ((.)OH) but not superoxide (O(.-)(2)) free radicals and that hydrogen peroxide did not react with metformin. Nevertheless, when polymorphonuclear cells (PMN) are stimulated by phorbol myristate acetate (PMA), or above all by formyl methionine leucyl phenylalanine (fMLP), a systematic (although nonsignificant) decrease of the ROS-induced chimiluminescence (CL) was observed. These results suggest that metformin could directly scavenge ROS or indirectly act by modulating the intracellular production of superoxide anion, of which NADPH oxidase constitutes the major source. This could contribute to the additional benefits of metformin, especially those related to the improvement in the cardiovascular outcomes in diabetes. PMID- 12759889 TI - Synergistic activation of mitogen-activated protein kinase by insulin and adenosine triphosphate in liver cells: permissive role of Ca2+. AB - We have previously demonstrated that insulin and G(q)-coupled receptor agonists individually activate mitogen-activated protein kinase (MAPK) in liver cells and both effects involve an influx of extracellular Ca(2+). Yet, these agonists have opposing physiological actions on hepatocyte glucose metabolism. We thus investigated the interaction between insulin and the P2Y(2) purinergic agonist adenosine triphosphate (ATP) on MAPK in HTC cells, a model hepatocyte cell line, and determined the involvement of cytosolic Ca(2+). Insulin and ATP each induced a dose-dependent phosphorylation of p44/42 MAPK that was partially inhibited by EGTA. However, pretreatment with insulin markedly increased the MAPK phosphorylation response to ATP. This potentiation was canceled by chelation of extracellular Ca(2+) with EGTA. We used patch clamp electrophysiology and fluorescence microscopy to understand the role of intracellular Ca(2+) in this effect. Insulin and ATP, respectively, induced monophasic and multiphasic changes in membrane potential and intracellular Ca(2+) as expected. Pretreatment with 10 nmol/L insulin significantly decreased the initial rapid depolarization (inward nonselective cation current [NSCC]), as well as the compounded Ca(2+) response induced by 100 micro mol/L ATP. However, in Ca(2+)-free conditions, insulin did not modify the Ca(2+) mobilized from internal pools after stimulation with ATP. Upon Ca(2+) readmission, internal store depletion by ATP or thapsigargin doubled the rate of capacitative Ca(2+) influx, whereas insulin increased this influx 1.32-fold. On the other hand, insulin pretreatment counteracted the increased rate of Ca(2+) influx induced by ATP but not by thapsigargin. In summary, insulin counteracts the membrane potential and Ca(2+) responses to ATP in HTC cells. However, insulin and ATP effects on MAPK activation are synergistic and Ca(2+) influx plays a permissive role. Therefore, the opposing metabolic actions of insulin and G(q)-coupled receptor agonists involve an interaction in signaling pathways that resides downstream of Ca(2+) influx. PMID- 12759890 TI - Is there a threshold of visceral fat loss that improves the metabolic profile in obese postmenopausal women? AB - It is presently unclear how much visceral adipose tissue (VAT) loss is needed to induce favorable metabolic changes. Cross-sectional studies have proposed that a threshold level of VAT exceeding 110 cm(2) in women induces deleterious changes in the metabolic profile. It is presently unclear, however, if significant decreases in VAT below this given threshold significantly improve the metabolic profile more as compared to decreases that remain below 110 cm(2). To examine whether achieving versus not achieving the proposed VAT threshold impacts differently on the metabolic profile in postmenopausal women, we examined the effects of a VAT loss below the 110-cm(2) threshold versus those individuals who remained higher than 110 cm(2) after a weight loss program. Twenty-five sedentary obese (baseline % body fat, 47.7% +/- 4.1%; [mean +/- SD]) postmenopausal women aged between 51 and 71 years (59.7 +/- 5.6 years) and displaying high baseline levels of VAT accumulation (223 +/- 45 cm(2)) were submitted to a 1-year weight loss program with weight stabilization periods before and after weight reduction. Based on their loss of VAT after weight loss, subjects were characterized as "attainers" (post VAT levels < 110 cm(2); average, 96 +/- 10 cm(2); n = 10) or "non-attainers" (post VAT levels > 110 cm(2); average, 171 +/- 34 cm(2); n = 15). We compared changes in (1) plasma lipid-lipoprotein levels, (2) insulin sensitivity (euglycemic/hyperinsulinemic clamp), and (3) supine resting blood pressure between groups who achieved these 2 distinct levels of VAT. Attainers showed a 2-fold greater loss of VAT compared to non-attainers (-51.5% v -27.5%, P <.001). Attainers also showed a greater loss of body weight (-19.0% v -12.5%, P <.01) and fat mass (-34.8% v -18.4%, P <.001) after the program compared to non attainers. Despite significant differences in the loss of total fat and VAT after the weight loss program, attainers and non-attainers showed comparable improvements for plasma high-density lipoprotein-cholesterol (HDL-chol) levels (+62.5% v +50.0%, P = not significant [NS]), cholesterol/HDL-chol ratio (-45.5% v -36.5%, P = NS), insulin sensitivity (+34.1% v +23.2%, P = NS), and resting systolic (-6.9% v -5.1%, P = NS) and diastolic (-11.3% v -11.1%, P = NS) blood pressure. These results do not favor the idea that attaining levels of VAT below a threshold of 110 cm(2) is necessary to favorably improve the metabolic profile in obese postmenopausal women. Achieving or not the proposed threshold of VAT, independently of baseline values, appears to yield similar metabolic improvements in obese postmenopausal women. More moderate losses of VAT appear to yield similar metabolic improvements as large losses. PMID- 12759891 TI - Elevated levels of interleukin-18 and tumor necrosis factor-alpha in serum of patients with type 2 diabetes mellitus: relationship with diabetic nephropathy. AB - To compare levels of interleukin (IL)-18, tumor necrosis factor-alpha (TNF alpha), and IL-6 in serum, we studied 151 type 2 diabetes mellitus patients with various degrees of nephropathy, as well as 80 healthy volunteers. IL-18, TNF alpha, and IL-6 in serum were measured using an enzyme-linked immunosorbent assay (ELISA) with the respective mouse monoclonal antibodies. Significant differences in serum levels of IL-18 and TNF-alpha were observed between the patients and control subjects (IL-18, 278.0 +/- 11.9 pg/mL v 172.8 +/- 7.7 pg/mL, P <.0001; TNF-alpha, 2.41 +/- 0.18 pg/mL v 0.46 +/- 0.18 pg/mL, P <.0001), whereas that of IL-6 was not different between the two groups (0.73 +/- 0.10 pg/mL v 0.65 +/- 0.08 pg/mL, difference not significant [NS]), although patients with nephropathy showed higher levels. In addition, IL-18 levels were increased in diabetic patients with the development of urinary albumin excretion, with the highest found in those with microalbuminuria (<30 micro g/mg creatinine, 252.7 +/- 16.4 pg/mL; 30 to >300 micro g/mg creatinine, 352.7 +/- 35.2 pg/mL; >>300 micro g/mg creatinine, 350.0 +/- 16.0 pg/mL). Similarly, TNF-alpha and IL-6 in diabetic patients with microalbuminuria or clinical albuminuria were significantly increased as compared with those without albuminuria (TNF-alpha, 3.20 +/- 0.41 pg/mL v 1.94 +/- 0.18 pg/mL; IL-6, 1.64 +/- 1.11 pg/mL v 0.51 +/- 0.05 pg/mL, P <.05, respectively). These results suggest that serum levels of IL-18, TNF-alpha, and IL-6 may have some etiopathogenic roles in diabetic nephropathy. PMID- 12759892 TI - Experimental model of postprandial hypertriglyceridemia in sucrose-fed rats and the effectiveness of atorvastatin in the model. AB - Although postprandial hypertriglyceridemia has drawn attention as an independent risk factor of cardiovascular disease, there is no established animal model that shows a physiological transitory change in lipoprotein metabolism after ingestion of a fatty meal. We developed an animal model of postprandial hypertriglyceridemia using sucrose-fed rats, and used this model to evaluate the effect of atorvastatin on this condition. Compared with normal rats, sucrose-fed rats orally loaded with olive oil showed a high and prolonged increase in plasma triglyceride (TG) concentration accompanied by both an increase in TG secretion and decrease in TG clearance. Atorvastatin (30 mg/kg orally) for 2 weeks reduced not only fasting plasma TG concentration, but also the postprandial TG concentration. Atorvastatin also suppressed rates of TG secretion in both chylomicron (CM)-rich (d < 0.96 g/mL) and very-low-density lipoprotein (VLDL) (d = 0.96 to 1.006 g/mL) fractions after oral fat loading. Further, atorvastatin improved the elimination time of exogenous TG emulsion only in the nonfasted, namely, high plasma TG condition. These results indicate that this animal model satisfactorily replicates the postprandial hypertriglyceridemia observed in humans and may therefore be useful in evaluation of lipid-lowering agents. Furthermore, atorvastatin not only improves fasting but also postprandial lipoprotein metabolism, presumably by reducing TG secretion from the liver or intestine or both, and by secondarily increasing TG-rich lipoprotein clearance by eliminating saturation. PMID- 12759893 TI - Hypertriglyceridemia is associated with increased insulin resistance in subjects with normal glucose tolerance: evaluation in a large cohort of subjects assessed with the 1999 World Health Organization criteria for the classification of diabetes. AB - The current study retrospectively examined the association between insulin resistance and plasma triglycerides (TG) in a group of subjects with normal glucose tolerance. Among 1,434 subjects consecutively undergoing a standard oral glucose tolerance test (OGTT) between 1993 and 1998, 567 (age, 15 to 78 years) were classified as having a normal glucose tolerance according to the 1999 World Health Organization (WHO) criteria and were selected for the study. Serum insulin was measured by radioimmunoassay (INSI-CTK, Dia Sorin, Saluggia, Italy). Intra assay and interassay coefficients of variation for the method were less than 4% and less than 8.5%, respectively. Insulin resistance was calculated by a homeostasis model assessment (HOMA(IR) = fasting serum insulin [mU/mL] x fasting blood glucose [mmol/L]/22.5). A very significant correlation was found between HOMA(IR) and plasma TG (r = 0.27, P < 1.02E(-10)). Multiple regression analyses confirmed plasma TG as independent variables explicative of HOMA(IR). When subjects were evaluated according to tertiles of TG, those in the upper two tertiles were older (P <.001) and presented higher body mass index (BMI) values (P <.0001) in comparison to subjects in the lower tertile. A positive trend (analysis of variance [ANOVA]) was found in regard to systolic (P <.05) and diastolic blood pressure (P <.0001), fasting blood glucose (P <.01), fasting serum insulin (P <.0001), and total cholesterol (P <.0001), while a negative trend was found in regard to high-density lipoprotein cholesterol (HDL-C) (P <.0001). Insulin resistance, calculated as HOMA(IR), was higher in the upper two tertiles of TG in comparison to the lower tertile (P <.001 and P <.0001, respectively), with a statistically significant trend for the entire group (first tertile, 1.85 +/- 0.94; second tertile, 2.28 +/- 1.10; third tertile, 2.65 +/- 1.71; ANOVA: P <.0001). In conclusion, this study shows an association between high levels of circulating TG and insulin resistance in patients with normal glucose tolerance seen in an atherosclerosis prevention clinic. This association is also present at levels of plasma TG considered to be normal and is associated with a cluster of cardiovascular risk factors. PMID- 12759894 TI - Total energy expenditure and carbohydrate oxidation are increased in the human immunodeficiency virus lipodystrophy syndrome. AB - To determine whether total energy expenditure (TEE) is increased in the human immunodeficiency virus (HIV) lipodystrophy syndrome, we compared energy expenditure (EE) and substrate oxidation rates in 12 HIV-infected men with lipodystrophy, 7 HIV-infected men without lipodystrophy, and 14 healthy controls. TEE and nutrient oxidation rates were assessed by whole-room indirect calorimetry. Resting energy expenditure (REE) was measured by indirect calorimetry using the open-circuit technique. Body composition was assessed by dual-energy x-ray absorptiometry (DEXA). Insulin sensitivity was measured using the insulin-modified frequently sampled intravenous glucose tolerance test. TEE adjusted for lean body mass (LBM) was significantly higher in the HIV-infected group with lipodystrophy compared to HIV-infected patients without lipodystrophy (2,873.3 +/- 69 v 2,573.9 +/- 92 kcal/d, P =.02) and compared to healthy controls (2,873.3 +/- 69 v 2,404.0 +/- 64 kcal/d, P <.001). REE and sleeping metabolic rate (SMR) adjusted for LBM were also significantly higher in the HIV-infected group with lipodystrophy compared to both HIV-infected and healthy controls. Carbohydrate oxidation rates adjusted for LBM were higher in men with HIV lipodystrophy as compared to healthy controls (362.5 +/- 23 v 250.0 +/- 22 g/d, P = <.01) and tended to be higher as compared to HIV-infected controls (362.5 +/- 23.6 v 297.3 +/- 31 g/d, P =.1). In conclusion, TEE and carbohydrate oxidation are increased in the HIV lipodystrophy syndrome. The increase in TEE appears to be due to increases in REE. The pathogenesis of elevated EE in HIV lipodystrophy and other forms of lipodystrophy remains to be determined. PMID- 12759895 TI - Exchangeable magnesium pool masses in spontaneously hypertensive rats. AB - We explored magnesium (Mg) metabolism by determination of exchangeable Mg pool masses and Mg kinetic parameters using stable Mg isotopes in spontaneously hypertensive rats (SHRs). Classical intracellular and extracellular Mg status biomarkers were also measured. Male SHRs and their male Wistar Kyoto (WKY) controls were fed a semipurified diet containing Mg 550 mg /kg for 2 weeks. Each rat received then an intravenous injection of 1.37 mg (25)Mg. The plasma (25)Mg disappearance curve over the next 7 days was used to measure the mass and fractional transport rate of 3 rapidly exchanging Mg metabolic pools, M1, M2, and M3. In the SHRs, plasma and erythrocyte Mg levels and urinary Mg excretion were not modified compared with their control WKYs, but tibia Mg level was significantly lower in the SHRs. Pool M3, the deep tissue pool, was significantly lower in SHRs compared with WKYs, but pools M1 and M2, the extracellular Mg pools, were statistically similar. The fractional transport rate of Mg from M1 to M2 and from M2 to M1 in the SHRs was higher than in the controls. The half-life of M1 was significantly decreased in SHRs compared with WKYs. In conclusion, this work demonstrates a decrease in intracellular Mg stores in SHRs compared with WKYs and disturbance of Mg exchanges in extracellular Mg, confirming a Mg metabolism disturbance in spontaneously hypertensive rats. Further work is now needed to elucidate the origin of the Mg depletion in SHRs and to explore Mg pools in hypertensive patients. PMID- 12759896 TI - Meal-induced 24-hour profile of circulating glycated insulin in type 2 diabetic subjects measured by a novel radioimmunoassay. AB - Increasing evidence supports a role for glycated insulin in the insulin-resistant state of type 2 diabetes. We measured 24-hour profiles of plasma glycated insulin, using a novel radioimmunoassay (RIA), to evaluate the effects of meal stimulation and intermittent fasting on circulating concentrations of plasma glycated insulin in type 2 diabetes. Patients (n = 6; hemoglobin A(1c) [HbA(1c)], 7.2% +/- 0.6%; fasting plasma glucose, 7.4 +/- 0.7 mmol/L; body mass index [BMI], 35.7 +/- 3.5 kg/m(2); age, 56.3 +/- 4.4 years) were admitted for 24 hours and received a standardized meal regimen. Half-hourly venous samples were taken for plasma glycated insulin, glucose, insulin, and C-peptide concentrations between 8 am and midnight and 2-hourly overnight. The mean plasma glycated insulin concentration over 24 hours was 27.8 +/- 1.2 pmol/L with a mean ratio of insulin:glycated insulin of 11:1. Circulating glucose, insulin, C-peptide, and glycated insulin followed a basal and meal-related pattern with most prominent increments following breakfast, lunch, and evening meal, respectively. The mean concentrations of glycated insulin during the morning, afternoon, evening, and night-time periods were 24.4 +/- 2.5, 28.7 +/- 2.3, 31.1 +/- 2.1, and 26.2 +/- 1.5 pmol/L, respectively, giving significantly higher molar ratios of insulin:glycated insulin of 18.0:1, 14.2:1, and 12.7:1 compared with 7.0:1 at night (P <.01 to P <.001). These data demonstrate that glycated insulin circulates at relatively high concentrations in type 2 diabetes with a diurnal pattern of basal and meal-stimulated release. A higher proportion of glycated insulin circulates at night suggestive of differences in metabolic clearance compared with native insulin. PMID- 12759897 TI - Regulation of oxysterol 7alpha-hydroxylase (CYP7B1) in the rat. AB - Cholesterol metabolized to 7alpha-hydroxylated bile acids is a principle pathway of cholesterol degradation. Cholesterol 7alpha-hydroxylase (CYP7A1) is the initial and rate-determining enzyme in the "classic pathway" of bile acid synthesis. An "alternative" pathway of bile acid synthesis begins with 27 hydroxylation of cholesterol by 27-hydroxylase (CYP27), followed by 7alpha hydroxylation by oxysterol 7alpha-hydroxylase (CYP7B1). The aim of the current study was to investigate the regulation of CYP7B1 by bile acids, cholesterol, and thyroid hormone in a previously well-studied in vivo model of bile acid synthesis, and to compare its regulation to that of CYP7A1. Three study groups were examined. In the first, male Sprague-Dawley rats with intact enterohepatic circulations were fed normal chow (controls), cholestyramine (CT), cholic acid (CA), chenodeoxycholic acid (CDCA), deoxycholic acid (DCA), or cholesterol (Chol). In the second group, taurocholate (TCA) was continuously intraduodenally infused for 48 hours to chronic biliary diverted rats. In a third set of studies, squalestatin, an inhibitor of cholesterol synthesis, was intravenously infused for 48 hours. In a fourth set of studies, the diurnal variation in CYP7B1 was compared to that of CYP7A1. At the end of each study livers were harvested, and CYP7B1 and CYP7A1 activities and mRNA levels were determined. Complete biliary diversion significantly increased the specific activity (SA) of both CYP7B1 ( upward arrow 212%; P <.002) and CYP7A1 ( upward arrow 212%; P <.007). Intraduodenal infusion of TCA to rats with biliary diversion decreased SA of both CYP7B1 ( downward arrow 29%; P <.001) and CYP7A1 ( downward arrow 46%; P <.01). The addition of CA, CDCA, or DCA to rat chow led to downregulation of CYP7B1 SAs by 42% (P <.003), 51% (P <.009), and 47% (P <.003), and CYP7A1 SAs by 32% +/- 6% (P <.003), 73% +/- 9% (P <.002), and 60% +/- 13% (P <.004), respectively. CT feeding upregulated both CYP7B1 ( upward arrow 136%; P <.004) and CYP7A1 ( upward arrow 216%; P <.001) SAs. While Chol feeding significantly upregulated CYP7A1 SA, no significant increase in CYP7B1 SA was found. Conversely, as previously shown in vitro, inhibition of cholesterol synthesis significantly suppressed both CYP7A1 and CYP7B1 activity and mRNA levels. Both CYP7B1 and CYP7A1 underwent diurnal variation, with peak and trough values for CYP7B1 lagging approximately 6 hours behind CYP7A1. We conclude that, in the rat, like CYP7A1, CYP7B1 demonstrates diurnal rhythm and is regulated by bile acids and cholesterol. PMID- 12759898 TI - The Leu7Pro polymorphism of the neuropeptide Y gene regulates free fatty acid metabolism. AB - The Leu7Pro polymorphism in the signal peptide of the preproneuropeptide Y (NPY) has been associated with dyslipidemias and free fatty acid (FFA) levels during exercise. The association of this polymorphism with insulin sensitivity has not been studied. In this study, the Leu7Pro polymorphism was determined in 2 groups of nondiabetic middle-aged subjects (n = 266 and n = 295). Insulin sensitivity was measured with the hyperinsulinemic euglycemic clamp (n = 266) or with an intravenous glucose tolerance test (IVGTT, n = 295). First-phase insulin secretion was determined as insulin area under the curve (AUC) during the first 10 minutes of the IVGTT. FFAs were measured both in the fasting state and during the hyperinsulinemic clamp. The Leu7Pro polymorphism of the NPY gene was not associated with the rates of whole body glucose uptake, insulin sensitivity index, insulin secretion during the IVGTT, or insulin AUC during the oral glucose tolerance test. However, the Pro7 allele was associated with low FFA levels both in the fasting state (P =.043) and during the hyperinsulinemic clamp (P =.003). In conclusion, the Leu7Pro polymorphism of the NPY gene associates with alterations in FFA metabolism but does not have an impact on insulin sensitivity, insulin secretion, or glucose metabolism. PMID- 12759899 TI - Comparison of 1-micro g and 250-micro g corticotropin stimulation tests for the evaluation of adrenal function in patients with acquired immunodeficiency syndrome. AB - Many patients with acquired immunodeficiency syndrome (AIDS) have symptoms suggestive of adrenal insufficiency, but a normal 250- micro g corticotropin (ACTH) stimulation test. We compared the results of 1- micro g and standard 250- micro g ACTH stimulation tests in patients with AIDS. Each patient was studied on 2 separate days. On day 1, 1 micro g ACTH was given intravenously at 8 am after an overnight fast and serum cortisol levels were measured at baseline, and 30 and 60 minutes after ACTH infusion. On day 2, the procedure was repeated with 250- micro g ACTH. An absolute peak cortisol value of > 18 micro g/dL and an increment of 7 micro g/dL or more from baseline constituted a normal response. Among 31 patients, 16 (52%) had discrepant results: 14 (45%) had subnormal responses to 1 micro g ACTH but normal responses to 250 micro g ACTH (group 1); 2 (6%) had normal responses to 1 micro g but subnormal responses to 250 micro g (group 2) ACTH; 6 patients (19%) had concordant abnormal responses (group 3); and 9 (30%) had concordant normal responses (group 4). Eight patients of group 1 underwent a confirmatory insulin tolerance test (ITT); 4 of these patients had abnormal responses to ITT. Kappa statistic and McNemar's test were used to evaluate the data. A kappa statistic value of 0.095 and a P value less than.003 for the McNemar test indicate only random level of agreement and significant differences in the probability of positive result between the 2 ACTH tests. We conclude that discrepancies between the 1- micro g and the 250- micro g ACTH stimulation tests are common in patients with AIDS, with the likelihood of agreement with the "gold standard" ITT of only 50% for each test in our sample of patients. Larger studies are needed to further evaluate the use of these tests in patients with AIDS. PMID- 12759900 TI - Differential effects of peroxisome proliferator-activated receptor activators on the mRNA levels of genes involved in lipid metabolism in primary human monocyte derived macrophages. AB - Peroxisome proliferator-activated receptors (PPARs) are key regulators of macrophage lipid metabolism. We compared the effects of three PPAR activators (bezafibrate, fenofibrate, and troglitazone) on the mRNA levels of genes involved in lipid metabolism in primary human macrophages and macrophage-derived foam cells. Treatment of human macrophages for 24 hours with 100 micro mol/L bezafibrate, a nonselective drug that activates the 3 PPAR subtypes (PPARalpha, PPARbeta/delta, and PPARgamma), caused an 87% (P <.01) and a 230% rise in CD36 and adipocyte fatty acid-binding protein (aP2) mRNA levels, respectively, whereas the expressions of PPARgamma, PPARalpha, acyl-CoA oxidase, carnitine palmitoyltransferase I (CPT-I), adenosine triphosphate (ATP)-binding cassette transporter 1 (ABCA1), neutral cholesteryl ester hydrolase, and lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) were not modified. However, treatment with selective PPARalpha (fenofibrate at 100 micro mol/L) and PPARgamma (troglitazone at 5 micro mol/L) activators had different effects. Fenofibrate increased PPARalpha (62%, P <.05) and LOX-1 (180%, P <.05) mRNA levels; and troglitazone upregulated CPT-I expression (75%, P <.05). When the effects of these drugs were assessed in macrophage-derived foam cells, we found that troglitazone caused a 134% (P <.05) and a 66% (P <.01) rise in ABCA1 and CPT-I mRNA levels, respectively, whereas the 3 drugs significantly increased aP2 transcripts (about 100% induction). Given that troglitazone treatment resulted in the upregulation of genes involved in the mitochondrial beta-oxidation of fatty acids (CPT-I) and in the reverse-cholesterol-transport pathway (ABCA1), we subsequently determined whether these changes affected intracellular cholesterol ester accumulation. In macrophage-derived foam cells a significant reduction (32%, P <.01) was observed in intracellular cholesterol accumulation after troglitazone, but not after bezafibrate or fenofibrate treatment. Since CPT-I inhibition promotes cholesterol incorporation into cholesteryl esters in macrophages, study is now needed on whether CPT-I induction by troglitazone may reduce the availability of fatty acids for synthesizing cholesterol esters, leading to less foam cell formation. PMID- 12759901 TI - Development of new pseudopotential methods: improved model core potentials for the first-row transition metals. AB - We have recently developed new nonrelativistic and scalar-relativistic pseudopotentials for the first-row transition metal and several main-group elements. These improved Model Core Potentials were tested on a variety of transition metal complexes to determine their accuracy in reproducing electronic structures, bond lengths, and harmonic vibrational frequencies with respect to both all-electron reference data as well as experimental data. The new potentials are also compared with the previous model core potentials available for the first row transition metals. The new potentials do a superior job at reproducing atomic data, reproduce molecular data as well as the previous version, and in conjunction with new main-group pseudopotentials that have L-shell structure of the valence basis set, they are slightly faster. PMID- 12759903 TI - Peptide models. XXXIII. Extrapolation of low-level Hartree-Fock data of peptide conformation to large basis set SCF, MP2, DFT, and CCSD(T) results. The Ramachandran surface of alanine dipeptide computed at various levels of theory. AB - At the dawn of the new millenium, new concepts are required for a more profound understanding of protein structures. Together with NMR and X-ray-based 3D structure determinations in silico methods are now widely accepted. Homology based modeling studies, molecular dynamics methods, and quantum mechanical approaches are more commonly used. Despite the steady and exponential increase in computational power, high level ab initio methods will not be in common use for studying the structure and dynamics of large peptides and proteins in the near future. We are presenting here a novel approach, in which low- and medium-level ab initio energy results are scaled, thus extrapolating to a higher level of information. This scaling is of special significance, because we observed previously on molecular properties such as energy, chemical shielding data, etc., determined at a higher theoretical level, do correlate better with experimental data, than those originating from lower theoretical treatments. The Ramachandran surface of an alanine dipeptide now determined at six different levels of theory [RHF and B3LYP 3-21G, 6-31+G(d) and 6-311++G(d,p)] serves as a suitable test. Minima, first-order critical points and partially optimized structures, determined at different levels of theory (SCF, DFT), were completed with high level energy calculations such as MP2, MP4D, and CCSD(T). For the first time three different CCSD(T) sets of energies were determined for all stable B3LYP/6 311++G(d,p) minima of an alanine dipeptide. From the simplest ab initio data (e.g., RHF/3-21G) to more complex results [CCSD(T)/6-311+G(d,p)//B3LYP/6 311++G(d,p)] all data sets were compared, analyzed in a comprehensive manner, and evaluated by means of statistics. PMID- 12759902 TI - Development of polyphosphate parameters for use with the AMBER force field. AB - Accurate force fields are essential for reproducing the conformational and dynamic behavior of condensed-phase systems. The popular AMBER force field has parameters for monophosphates, but they do not extend well to polyphorylated molecules such as ADP and ATP. This work presents parameters for the partial charges, atom types, bond angles, and torsions in simple polyphosphorylated compounds. The parameters are based on molecular orbital calculations of methyldiphosphate and methyltriphosphate at the RHF/6-31+G* level. The new parameters were fit to the entire potential energy surface (not just minima) with an RMSD of 0.62 kcal/mol. This is exceptional agreement and a significant improvement over the current parameters that produce a potential surface with an RMSD of 7.8 kcal/mol to that of the ab initio calculations. Testing has shown that the parameters are transferable and capable of reproducing the gas-phase conformations of inorganic diphosphate and triphosphate. Also, the parameters are an improvement over existing parameters in the condensed phase as shown by minimizations of ATP bound in several proteins. These parameters are intended for use with the existing AMBER 94/99 force field, and they will permit users to apply AMBER to a wider variety of important enzymatic systems. PMID- 12759904 TI - Powder pattern indexing using the weighted crosscorrelation and genetic algorithms. AB - X-ray diffraction is a powerful technique for investigating the structure of crystals and crystalline powders. Unfortunately, for powders, the first step in the structure elucidation process, retrieving the unit cell parameters (indexing), is still very critical. In the present article, an improved approach to powder pattern indexing is presented. The proposed method matches peak positions from experimental X-ray powder patterns with peak positions from trial cells using a recently published method for pattern comparison (weighted crosscorrelation). Trial cells are optimized with Genetic Algorithms. Patterns are not pretreated to remove any existing zero point shift, as this is determined during optimization. Another improvement is the peak assignment procedure. This assignment is needed for determining the similarity between lines from trial cells and experiment. It no longer allows calculated peaks to be assigned twice to different experimental peaks, which is beneficial for the indexing process. The procedure proves to be robust with respect to false peaks and accidental or systematic absensences of reflections, and is successfully applied to powder patterns originating from orthorhombic, monoclinic, and triclinic compounds measured with synchrotron as well as with conventional laboratory X-ray diffractometers. PMID- 12759905 TI - Methyl dynamics in crystalline amino acids: MD and NMR. AB - Correlation times for rotation of deuterated methyls in crystalline leucine, valine, and cyclo-L-alanyl-L-alanine are calculated with molecular dynamics and compared with NMR data. The simulations distinguish between methyls having different steric environments in the crystal, yielding correlation times differing by a factor of up to 30 for methyls within a given crystal. MD and NMR correlation times agree to within a factor of 2. However, averaging over nonequivalent methyls can yield correlation functions that, although actually multiexponential, are well fit by single exponentials. This may have significance for interpreting NMR data; previous NMR data did not distinguish between the methyls in these crystals. Adiabatic rotational barriers calculated with the X ray structure differ from effective barriers during simulation by up to +/-1 kcal/mol; the difference indicates that dynamical effects have a significant role in determining rotational correlation times. PMID- 12759907 TI - A local interpolation scheme using no derivatives in potential sampling: application to O(1D) + H2 system. AB - We recently proposed a local interpolation scheme, in which interpolant moving least squares (IMLS) and Shepard interpolation are employed to describe potential energy surfaces. This IMLS/Shepard scheme is used to interpolate quantum chemical potential energy surfaces for which analytical derivatives are not available. In this study, we apply the scheme to the highly exothermic O((1)D) + H(2) --> H + OH reaction and compare it with results based on Shepard interpolation using second-order Taylor expansions. An analytical surface is used to define the potential function so that errors in the interpolation function may accurately be determined. We find that the present scheme reproduces the correct reactive cross sections more accurately than the Shepard scheme, and with rms errors for energy and gradients that are significantly smaller than those from Shepard interpolation. This occurs even though the present scheme does not utilize derivative and Hessian information, whereas the Shepard interpolation does. The Bayesian approach proposed by Bettens and Collins does not improve the IMLS/Shepard results significantly, although it does the Shepard-only approach. The accuracy of the IMLS/Shepard scheme is surprising, but can be explained by the more global nature of the interpolation. PMID- 12759908 TI - A novel approach for designing simple point charge models for liquid water with three interaction sites. AB - A simultaneous improvement of the diffusion and dielectric properties of the simple point charge (SPC) model for liquid water appears to be very difficult with conventional reparametrization of the commonly used Lennard-Jones and Coulomb interaction functions and without including a self-energy correction in the effective pair-potential as is done in the SPC/E model. Here, a different approach to circumvent this problem is presented. A short-range interaction term, which corrects the oxygen-oxygen energy at small distances by small amounts of energy, was introduced in the nonbonded interaction function. This additional force-field term allows to derive new parameter sets for SPC-like water models that yield better agreement with experimental data on liquid water. Based on previous investigations of the force-field parameter dependence of the water properties of SPC-like models, the necessary parameter changes to obtain a lower diffusion coefficient and a larger dielectric permittivity were specified and accordingly six new models were developed. They all represent an improvement over SPC in terms of structural and diffusional properties, four of them show better dielectric properties also. One model, SPC/S, has been characterized in more detail, and represents most properties of liquid water better than SPC while avoiding the larger discrepancies with experimental values regarding density, thermal compressibility, energy, and free energy of the SPC/E model. We conclude that the use of a simple, short-ranged additional oxygen-oxygen interaction term makes a simultaneous improvement of the diffusion coefficient and the dielectric properties of water feasible. PMID- 12759906 TI - An extensible and systematic force field, ESFF, for molecular modeling of organic, inorganic, and organometallic systems. AB - ESFF is a rule-based force field designed for modeling organic, inorganic, and organometallic systems. To cover this broad range of molecular systems, ESFF was developed in an extensible and systematic manner. Several unique features were introduced including pseudoangle and a dot product function representing torsion energy terms. The partial atomic charges that are topology-dependent are determined from ab initio (DFT) calculated electronegativity and hardness for valence orbitals. The van der Waals parameters are charge-dependent, and correlated with the ionization potential for atoms in various valence states. To obtain a set of well-defined and physically meaningful parameters, ESFF employs semiempirical rules to translate atomic-based parameters to parameters typically associated with a covalent valence force field. The atomic parameters depend not only on atom type, but also on internal type, thus resulting in a more accurate force field. This article presents the theory and the method used to develop the force field. The force field has been applied to molecular simulations of a wide variety of systems including nucleic acids, peptides, hydrocarbons, porphyrins, transition metal complexes, zeolites, and organometallic compounds. Agreement with the experimental results indicates that ESFF is a valuable tool in molecular simulations for understanding and predicting both crystal and gas phase molecular structures. PMID- 12759909 TI - Complete graph conjecture for inner-core electrons: homogeneous index case. AB - The complete graph conjecture that encodes the inner-core electrons of atoms with principal quantum number n >or= 2 with complete graphs, and especially with odd complete graphs, is discussed. This conjecture is used to derive new values for the molecular connectivity and pseudoconnectivity basis indices of hydrogen suppressed chemical pseudographs. For atoms with n = 2 the new values derived with this conjecture are coincident with the old ones. The modeling ability of the new homogeneous basis indices, and of the higher-order terms, is tested and compared with previous modeling studies, which are centered on basis indices that are either based on quantum concepts or partially based on this new conjecture for the inner-core electrons. Two similar algorithms have been proposed with this conjecture, and they parallel the two "quantum" algorithms put forward by molecular connectivity for atoms with n > 2. Nine properties of five classes of compounds have been tested: the molecular polarizabilities of a class of organic compounds, the dipole moment, molar refraction, boiling points, ionization energies, and parachor of a series of halomethanes, the lattice enthalpy of metal halides, the rates of hydrogen abstraction of chlorofluorocarbons, and the pED(50) of phenylalkylamines. The two tested algorithms based on the odd complete graph conjecture give rise to a highly interesting model of the nine properties, and three of them can even be modeled by the same set of basis indices. Interesting is the role of some basis indices all along the model. PMID- 12759911 TI - Hydrogen bonding in diols and binary diol-water systems investigated using DFT methods. II. Calculated infrared OH-stretch frequencies, force constants, and NMR chemical shifts correlate with hydrogen bond geometry and electron density topology. A reevaluation of geometrical criteria for hydrogen bonding. AB - Although the two hydroxyl groups in 1,2-diols interact as evidenced by NMR and IR spectroscopic shifts, electron density topological analysis has shown a bond critical point (BCP) and atomic bond path to be absent (Klein, R. A.; J Comp Chem 2002, 23, 585-599; J Am Chem Soc 2002, 124, 13931-13937), indicating that no intramolecular hydrogen bond is formed. Here, we demonstrate that small NMR or IR shifts are neither necessarily diagnostic nor sufficient as indicators of hydrogen bond formation; moreover, modified van der Waals atomic radii are needed for estimating maximum nuclear interaction distances and nuclear interpenetration. PMID- 12759910 TI - A density functional study of the hydrogen-bond network within the HIV-1 protease catalytic site cleft. AB - The relative energy between two different protonation sites of the Asp25' catalytic site residue is computed and analyzed for various HIV-1 Protease/inhibitor complexes and compared to the wild-type structure. By comparing calculations of negatively charged fragments of gradually increasing size up to 105 atoms we show that correct modeling of the HIV-1 Protease active site requires much larger models than the commonly used acetic acid/acetate moieties. The energy difference between the two proposed protonation sites decreases as the size of the system increases and tends to converge only when the entire catalytic triad of both monomers is taken into account. The importance of the Gly27 backbone amine groups in the stabilization of the negative charge within the catalytic site cleft is revealed. Comparison of the wild-type structure with the structures from various Pr/drug complexes indicates that the HIV-1 protease has a particular catalytic site flexibility. PMID- 12759912 TI - Localized basis orbitals: minimization of 2-electron integrals array and orthonormality of basis set. AB - A new scheme for deriving localized basis orbitals (LBOs) and for obtaining integral transformations from the basis orbitals (BOs) to the LBOs has been introduced. The scheme was tested at the ab initio Hartree-Fock level using the STO-3G basis set. It has been revealed that it provides results that are close to the conventional ab initio approximations for various physical-chemical properties. At the same time, both the number of differential overlaps and the number of electron repulsion integrals (ERIs) grow with the system size notably slower than those calculated for the usual BOs. The power exponent for ERI/LBO is typically smaller by 0.3-0.6 than that for ERI/BO. The exponent reaches the value of 1.69 even for triglycine (24 atoms only), which represents a relatively small molecular model. Thus, the localization of the BOs (using LBOs) may result in additional improvements in efficiency even for electronically delocalized systems. It was shown that ERI/LBO is particularly efficient for systems with complex spatial structures (including conjugated species). The obtained results indicate that the proposed scheme could be included in computational methods targeted at calculating large molecular systems (which achieve linear scaling for more distant interactions). Neglecting ERI/LBO does not depend on the delocalization of the localized MO using ERI/LBO. The orthogonality and locality of the LBOs should make them useful in methods based on dividing the system into orthogonal subsystems. PMID- 12759913 TI - Optimized Slater-type basis sets for the elements 1-118. AB - Seven different types of Slater type basis sets for the elements H (Z = 1) up to E118 (Z = 118), ranging from a double zeta valence quality up to a quadruple zeta valence quality, are tested in their performance in neutral atomic and diatomic oxide calculations. The exponents of the Slater type functions are optimized for the use in (scalar relativistic) zeroth-order regular approximated (ZORA) equations. Atomic tests reveal that, on average, the absolute basis set error of 0.03 kcal/mol in the density functional calculation of the valence spinor energies of the neutral atoms with the largest all electron basis set of quadruple zeta quality is lower than the average absolute difference of 0.16 kcal/mol in these valence spinor energies if one compares the results of ZORA equation with those of the fully relativistic Dirac equation. This average absolute basis set error increases to about 1 kcal/mol for the all electron basis sets of triple zeta valence quality, and to approximately 4 kcal/mol for the all electron basis sets of double zeta quality. The molecular tests reveal that, on average, the calculated atomization energies of 118 neutral diatomic oxides MO, where the nuclear charge Z of M ranges from Z = 1-118, with the all electron basis sets of triple zeta quality with two polarization functions added are within 1-2 kcal/mol of the benchmark results with the much larger all electron basis sets, which are of quadruple zeta valence quality with four polarization functions added. The accuracy is reduced to about 4-5 kcal/mol if only one polarization function is used in the triple zeta basis sets, and further reduced to approximately 20 kcal/mol if the all electron basis sets of double zeta quality are used. The inclusion of g-type STOs to the large benchmark basis sets had an effect of less than 1 kcal/mol in the calculation of the atomization energies of the group 2 and group 14 diatomic oxides. The basis sets that are optimized for calculations using the frozen core approximation (frozen core basis sets) have a restricted basis set in the core region compared to the all electron basis sets. On average, the use of these frozen core basis sets give atomic basis set errors that are approximately twice as large as the corresponding all electron basis set errors and molecular atomization energies that are close to the corresponding all electron results. Only if spin-orbit coupling is included in the frozen core calculations larger errors are found, especially for the heavier elements, due to the additional approximation that is made that the basis functions are orthogonalized on scalar relativistic core orbitals. PMID- 12759914 TI - Estimation of the determinants of household health care expenditures in Nepal with controls for endogenous illness and provider choice. AB - This paper uses the Nepal Living Standards Survey, a nationally representative sample of households from 1996, to investigate the determinants of household out of-pocket health expenditures. The analysis uses a multi-equation joint estimation to control for endogeneity of sickness and provider choice. The results of this analysis indicate several interesting findings. First, common unobserved factors were found to be statistically significant determinants of illness, choice of provider, and health expenditures, and may cause bias to parameter estimates if not controlled. Second, the income elasticity is estimated to be 1.10, with income having both a direct effect on health expenditure, and an indirect effect through likelihood of illness and the type of provider that is chosen. Third, housing and sanitary conditions were found to have a substantial effect on illness, and as a result, out-of-pocket health care expenditures. Fourth, despite the fact that urban, ill individuals who seek care are more likely to utilize care in more expensive settings, average health care expenditure among the urban sample was found to be substantially lower than among the rural sample, partly due to a lower likelihood of reporting illnesses and injuries and of using any type of health care provider. PMID- 12759915 TI - A preliminary investigation of the effects of restrictions on Medicaid funding for abortions on female STD rates. AB - There is evidence in the economics literature that restrictions on Medicaid funding for abortion reduces the demand for abortion. The unresolved question is whether such restrictions also increase safe sex (that is, pregnancy avoidance) behavior among women. This study explores that issue using state-level gonorrhea rates among women for 1975-1995. The rationale is that sexual behavior that leads to greater risk of accidental pregnancies is likely to be highly correlated with sexual behavior leading to greater risk of STD infection. Since gonorrhea has an incubation period of about a week, and is transmitted almost exclusively through sexual intercourse, a change in sexual behavior should soon be followed by a change in gonorrhea rates. The study used a partial adjustment model with lagged dependent variables estimated using Arellano-Bond's GMM method. Results fail to find any statistically significant evidence that Medicaid funding restrictions are effective in reducing gonorrhea rates. This finding is robust to a variety of alternate specifications and tests. This suggests that restrictions on Medicaid funding for abortion fail to promote safe sex behavior among women. PMID- 12759916 TI - The labour market for nursing: a review of the labour supply literature. AB - The need to ensure adequate numbers of motivated health professionals is at the forefront of the modernisation of the UK NHS. The aim of this paper is to assess current understanding of the labour supply behaviour of nurses, and to propose an agenda for further research. In particular, the paper reviews American and British economics literature that focuses on empirical econometric studies based on the classical static labour supply model. American research could be classified into first generation, second generation and recent empirical evidence. Advances in methods mirror those in the general labour economics literature, and include the use of limited dependent variable models and the treatment of sample selection issues. However, there is considerable variation in results, which depends on the methods used, particularly on the effect of wages. Only one study was found that used UK data, although other studies examined the determinants of turnover, quit rates and job satisfaction. The agenda for further empirical research includes the analysis of discontinuities in the labour supply function, the relative importance of pecuniary and non-pecuniary job characteristics, and the application of dynamic and family labour supply models to nursing research. Such research is crucial to the development of evidence based policies. PMID- 12759917 TI - Conjoint analysis. The cost variable: an Achilles' heel? AB - This paper seeks to enlighten the readers on the potential complexities involved in including cost variables in conjoint analysis, with the aim of emphasising that interpretation of implicit WTP values should be tackled with caution. To illustrate the potential pitfalls, a large data set from a recent Danish study is applied. The data consists of 1991 interviews in which participants are required to perform three discrete choice tasks regarding choice of hospitals, and three choice tasks involving health-care systems in general. Model comparisons are performed which test the effect of (1) the cost range applied and (2) the effect of including a dummy variable to represent the utility associated with payment per se. A wider cost range including higher payments is associated with lower parameter weights associated with the payment variable, and thus increased WTP values. Including a dummy variable to explain utility associated with payment per se has significant effects on the model incurring some of the other variables to become insignificant, and others to change sign. Results suggest that inclusion of a two-dimensional structure to explain the relationship between cost and utility may avoid erroneous conclusions and give rise to significant changes in implicit WTP estimates. PMID- 12759918 TI - The efficiency of health production: re-estimating the WHO panel data using parametric and non-parametric approaches to provide additional information. AB - The World Health Report 2000 focuses on the performance of health-care systems around the globe. The report uses efficiency measurement techniques to create a league table of health-care systems, highlighting good and bad performers. Efficiency is measured using panel data methods. This paper suggests that the WHO's estimation procedure is too narrow and that contextual information is hidden by the use of one method. This paper uses and validates a range of parametric and non-parametric empirical methods to measure efficiency using the WHO data. The rankings obtained are compared to the WHO league table and we demonstrate that there are trends and movements of interest within the league tables. We recommend that the WHO broaden its range of techniques in order to reveal this hidden information. PMID- 12759919 TI - The person trade-off method and the transitivity principle: an example from preferences over age weighting. AB - The person trade-off (PTO) is increasingly being used to elicit preferences in health. This paper explores the measurement properties of the PTO method in the context of a study about how members of the public prioritise between patients of different ages. In particular, it considers whether PTO responses satisfy the transitivity principle; that is, whether one PTO response can be inferred from two other PTO responses. The results suggest that very few responses to PTO questions satisfy cardinal transitivity condition. However, this study has produced results that suggest that cardinal transitivity will hold, on average, when respondents who fail to satisfy the ordinal transitivity condition have been excluded from the analysis. This suggests that future PTO studies should build in checks for ordinal transitivity. PMID- 12759920 TI - A note on the decomposition of the health concentration index. AB - In recent work, the concentration index has been widely used as a measure of income-related health inequality. The purpose of this note is to illustrate two different methods for decomposing the overall health concentration index using data collected from a Short Form (SF-36) survey of the general Australian population conducted in 1995. For simplicity, we focus on the physical functioning scale of the SF-36. Firstly we examine decomposition 'by component' by separating the concentration index for the physical functioning scale into the ten items on which it is based. The results show that the items contribute differently to the overall inequality measure, i.e. two of the items contributed 13% and 5%, respectively, to the overall measure. Second, to illustrate the 'by subgroup' method we decompose the concentration index by employment status. This involves separating the population into two groups: individuals currently in employment; and individuals not currently employed. We find that the inequality between these groups is about five times greater than the inequality within each group. These methods provide insights into the nature of inequality that can be used to inform policy design to reduce income related health inequalities. PMID- 12759921 TI - Genome-wide amplification and allelotyping of sporadic pituitary adenomas identify novel regions of genetic loss. AB - Through the use of a candidate gene approach, several previous studies have identified loss of heterozygosity (LOH) at putative tumor-suppressor gene (TSG) loci in sporadic pituitary tumors. This study reports a genome-wide allelotyping by use of 122 microsatellite markers in a large cohort of tumors, consisting of somatotrophinomas and non-functioning adenomas. Samples were first subject to prior whole genome amplification by primer extension pre-amplification (PEP) to circumvent limitations imposed by insufficient DNA for whole-genome analysis with this number of microsatellite markers. The overall mean frequency of loss in invasive tumors was significantly higher than that in their non-invasive counterparts (7 vs. 3% somatotrophinomas; 6 vs. 3% non-functioning adenomas, respectively). Analysis of the mean frequency of LOH, across all markers to individual chromosomal arms, identified 13 chromosomal arms in somatotrophinomas and 10 in non-functioning tumors, with LOH greater than the 99% upper confidence interval calculated for the rate of overall random allelic loss. In the majority of cases, these losses were more frequent in invasive tumors than in their non invasive counterparts, suggesting these to be markers of tumor progression. Other regions showed similar frequencies of LOH in both invasive and non-invasive tumors, implying these to be early changes in pituitary tumorigenesis. This genome-wide study also revealed chromosomal regions where losses were frequently associated with an individual marker, for example, chromosome arm 1q (LOH > 30%). In some cases, these losses were subtype-specific and were found at a higher frequency in invasive tumors than in their non-invasive counterparts. Identification of these regions of loss provides the first preliminary evidence for the location of novel putative TSGs involved in pituitary tumorigenesis that are, in some cases, subtype-specific. This investigation provides an unbiased estimate of global aberrations in sporadic pituitary tumors as assessed by LOH analysis. The identification of multiple "hotspots" throughout the genome may be a reflection of an unstable chromatin structure that is susceptible to a deletion or epigenetic-mediated gene-silencing events. PMID- 12759922 TI - Comparative analysis of MLL partial tandem duplication and FLT3 internal tandem duplication mutations in 956 adult patients with acute myeloid leukemia. AB - Partial tandem duplication (PTD) of the MLL gene and internal tandem duplication (ITD) of the juxtamembrane region of the FLT3 receptor tyrosine kinase gene have been described in acute myeloid leukemia (AML) patients, preferentially in those with normal cytogenetics. These alterations have been associated with a poor prognosis. In our study, we analyzed the prevalence and the potential prognostic impact of these aberrations in a large unselected and well-defined cohort of 956 patients with AML. Results were correlated with cytogenetic data and clinical outcome. MLL PTD was detected by RT-PCR, subsequent nucleotide sequencing, and Southern blotting. The overall incidence was found to be 5.0% (48/956), whereas FLT3 ITD was detected in 19.2% (184/956). Sixteen cases were positive for both alterations. The rate of MLL PTD in FLT3 ITD positive patients was significantly higher than that in FLT3 ITD negative patients [16/184 (8.7%); 32/772 (4.1%); P = 0.025]. However, both aberrations were highly increased in patients with normal karyotype (MLL PTD 35/431, P = 0.004; FLT3 ITD 132/334, P < 0.001). When restricted to this subgroup, the rate of MLL PTD in patients with FLT3 mutations was not significantly increased. No statistically significant differences were detected between patients positive for MLL PTD and patients negative for MLL PTD in the rate of complete remissions or the overall survival, although we did see a significantly shorter disease-free survival in patients age 60 or younger. In conclusion, although there is an overlap in the mutational spectrum in AML with FLT3 ITD and MLL PTD mutations, our data do not support a common mechanistic basis. Although associated with inferior disease-free survival, the results of this study do not unequivocally support the notion that MLL PTD mutations represent an independent prognostic factor. PMID- 12759923 TI - Patterns of aneuploidy in stage IV clear cell renal cell carcinoma revealed by comparative genomic hybridization and spectral karyotyping. AB - We report the use of spectral karyotyping (SKY) and comparative genomic hybridization (CGH) to describe the numerous genomic imbalances characteristic of stage IV clear cell renal cell carcinoma (CCRCC). SKY and CGH were performed on 10 cell lines established from nephrectomy specimens, and CGH on uncultured material from five of the primary renal tumors. The mutational status of VHL (3p25) and MET (7q31), genes implicated in renal carcinogenesis, were determined for each case. Each case showed marked aneuploidy, with an average number of copy alterations of 14.6 (+/-2.7) in the primary tumors and 19.3 (+/-4.6) in the cell lines. Both whole-chromosome and chromosome-segment imbalances were noted by CGH: consistent losses or gains included +5q23-->ter (100%), -3p14-->ter (80%), and +7 (70%). All VHL mutations and 83% of the genomic imbalances found in the primary tumors were also found in the cell lines derived from them. SKY showed many complex structural rearrangements that were undetected by conventional banding analysis in these solid tumors. All cases with VHL inactivation had 3p loss and 5q gain related primarily to unbalanced translocations between 3p and 5q. In contrast, gains of chromosome 7 resulted primarily from whole-chromosome gains and were not associated with mutations of MET. SKY and CGH demonstrated that genomic imbalances in advanced RCC were the result of either segregation errors [i.e., whole chromosomal gains and losses (7.8/case)] or chromosomal rearrangements (10.7/case), of which the majority were unbalanced translocations. PMID- 12759924 TI - Heterogeneous pattern of chromosomal breakpoints involving the MYC locus in multiple myeloma. AB - Chromosomal rearrangements of the MYC locus, which often involve the IG loci, are recurrent events in multiple myeloma (MM) and plasma cell leukemia (PCL). We used dual-color fluorescence in situ hybridization (FISH) to characterize the breakpoint locations of chromosomal translocations/rearrangements involving the MYC locus at 8q24 found in a panel of 14 MM cell lines and 70 primary tumors (66 MM and 4 PCL). MYC locus alterations were observed in 21 cases: MYC/IG (mainly IGH@) fusions in 11 cell lines and three patients (2 MM and 1 PCL), and extra signals and/or abnormal MYC localizations in seven patients (5 MM and 2 PCL). Fourteen of these cases were investigated by FISH analyses by use of a panel of BAC clones covering about 6 Mb encompassing the MYC locus. The breakpoints were localized in a region 100-250 kb centromeric to MYC in four cases, a region 500 800 kb telomeric to the gene in four cases, and regions > or = 2 Mb centromeric or telomeric to MYC in five cases. Two different breakpoints were detected in the KMS-18 cell line, whereas the insertion of a MYC allele was found in a complex t(16;22) chromosomal translocation in the RPMI8226 cell line. Our data document a relatively high dispersion of 8q24 breakpoints in MM. PMID- 12759925 TI - Cytogenetic, spectral karyotyping, fluorescence in situ hybridization, and comparative genomic hybridization characterization of two new secondary leukemia cell lines with 5q deletions, and MYC and MLL amplification. AB - Cytogenetic studies of patients with therapy-induced acute myeloid leukemia (t AML) have demonstrated whole chromosome loss or q-arm deletion of chromosomes 5 and/or 7 in a majority of cases. We have established two cell lines, SAML-1 and SAML-2, from two patients who developed t-AML after radiation and chemotherapy for Hodgkin disease. In both cases, the leukemia cells contained 5q deletions. SAML-1 has 58 chromosomes and numerous abnormalities, including der(1)(1qter- >1p22::5q31-->5qter), der(5)(5pter-->5q22::1p22-->1pter), +8, der(13)i(13)(q10)del(13)(q11q14.1), and t(10;11). Fluorescence in situ hybridization (FISH) with unique sequence probes for the 5q31 region showed loss of IL4, IL5, IRF1, and IL3, and translocation of IL9, DS5S89, EGR1, and CSFIR to 1p. SAML-2 has 45 chromosomes, del(5)(q11.2q31) with a t(12;13)ins(12;5), leading to the proximity of IRF1 and RB1, and complex translocations of chromosomes 8 and 11, resulting in amplification of MYC and MLL. Comparative genomic hybridization and spectral karyotyping were consistent with the G-banding karyotype and FISH analyses. Because a potential tumor suppressor(s) in the 5q31 region has yet to be identified, these cell lines should prove useful in the study of the mechanisms leading to the development of t-AML. PMID- 12759926 TI - High-resolution analysis of acquired genomic imbalances in bone marrow samples from chronic myeloid leukemia patients by use of multiple short DNA probes. AB - Chronic myeloid leukemia (CML) is a biphasic hematopoietic malignancy associated with a single cytogenetic aberration, the Philadelphia translocation t(9;22)(q34;q11), resulting in the BCR-ABL1 fusion oncogene. Molecular heterogeneity was recently demonstrated in the form of extensive deletion of chromosomes 9 and 22 material from the der(9)t(9;22) in 15% of CML patients. The deletions were associated with a worse disease prognosis. Further genetic heterogeneity is seen during the terminal blast crisis stage of CML, in the form of additional non-random chromosome abnormalities. These include most frequently an extra copy of the Ph chromosome, trisomy 8, and isochromosome 17q. We used the genetic heterogeneity of CML as a framework to explore a new technique for high throughput assessment of locus copy number in malignancy. Multiplex amplifiable probe hybridization (MAPH) relies on the ability of numerous short (100-300 bp) DNA probes to be recovered quantitatively by use of a common primer pair after hybridization to genomic DNA. Derivative chromosome 9 deletions were successfully mapped in a CML cell line (MC3) and nine patient bone marrow samples by simultaneous hybridization of 10 MAPH probes. All results were confirmed by fluorescence in situ hybridization. MAPH was found to be informative in the presence of up to 50% of normal cells, thus establishing the sensitivity of the technique in clonal tumor cell populations. MAPH was performed effectively on DNA samples extracted from fresh or methanol/acetic acid-fixed clonal cell populations. Amplifications of BCR-ABL1 were also detected and quantified in four CML cell lines by use of MAPH probes specific for ABL1 exon 11 and BCR exon 1. Our results demonstrate that MAPH is a reproducible high-throughput method suitable for the assessment of genomic imbalances of multiple loci in tumor DNA samples with heterogeneous cell populations at a resolution of 100-300 bp. PMID- 12759928 TI - SOCS1 methylation in patients with newly diagnosed acute myeloid leukemia. AB - The proliferation and differentiation of hematopoietic precursor cells depend on various cytokines. The suppressor of cytokine signaling-1 (SOCS1) down-regulates Janus kinases/signal transducers and activators of transcription (JAK/STAT) pathway activity and inhibits the biological effects of cytokines. SOCS1 has been shown to have tumor-suppressor activity, and methylation of this gene, resulting in transcriptional silencing, has been found in 65% of hepatocellular carcinoma and has been suggested to play an important role in the development of the cancer. The methylation status of the SOCS1 gene in acute myeloid leukemia (AML) has not been reported before. In this study, we analyzed SOCS1 methylation in 89 patients with newly diagnosed AML and correlated the result with immunophenotypes, cytogenetics, clinical features, and treatment outcome. SOCS1 methylation was found in the leukemic cells from 53 patients (60%). Thirteen (76%) of the 17 patients with t(15;17) had SOCS1 methylation, whereas this gene was methylated in only one (11%) of the nine patients with t(8;21). The frequencies of SOCS1 methylation among various cytogenetic subgroups differed significantly (P = 0.014). Other clinical and laboratory parameters and the disease-free survival and overall survival were similar between patients with and without SOCS1 methylation. In conclusion, SOCS1 methylation occurs in more than half of AML cases, correlates with cytogenetic abnormalities, and may play an important role in the development of subsets of AML. PMID- 12759927 TI - A study on the incidence of ABL gene deletion on derivative chromosome 9 in chronic myelogenous leukemia by interphase fluorescence in situ hybridization and its association with disease progression. AB - Fluorescence in situ hybridization for the BCR/ABL rearrangement in 138 bone marrow specimens from 59 Philadelphia(+) (Ph(+)) chronic myelogenous leukemia (CML) patients, 35 Ph(+) acute lymphoblastic leukemia (ALL) patients, and 57 Ph( ) ALL patients was used. Sixteen (27.1%) of the 59 CML patients had deletions of the residual ABL gene on the derivative chromosome 9. During the study period, 32 of the 59 CML patients progressed to blast crisis or accelerated phase. Of these, nine patients had residual ABL gene deletions on the derivative chromosomes 9 and 23 patients had no deletions. The mean duration from first diagnosis to blast crisis or accelerated phase for the nine patients with ABL deletions was 32.8 months, and for the 23 patients without ABL deletions, it was 62.4 months (P = 0.017). The overall survival time for the 16 patients with deletions was 32.8 months, and for the 43 patients without deletions, it was 60.1 months (P = 0.164). ABL deletions were not detected among the 35 ALL patients (17 with major BCR/ABL, 18 with minor BCR/ABL), and it appears that this deletion occurs rarely or not at all in Ph(+) ALL patients, which is in contrast to the CML patients (27.1%). However, we detected two ALL cases with ABL deletion but without BCR/ABL rearrangement among 49 Ph(-) ALL and 66 Ph(-) AML patients. In conclusion, patients with ABL deletions progress to blast crisis or accelerated phase in a significantly shorter time than do those without such deletions. It is therefore suggested that the ABL deletion is an indicator of a poor prognosis in CML. PMID- 12759929 TI - Haploinsufficiency and reduced expression of genes localized to the 8p chromosomal region in human prostate tumors. AB - Cytogenetic and molecular studies have suggested that deletion or rearrangement of sequences that map to the short arm of chromosome 8 may be permissive for tumorigenesis in several organ systems, and in human prostate tumors in particular. In this study, we hypothesized that genes deleted for one copy and localized to the 8p chromosomal region may be transcriptionally down-regulated or ablated in affected human prostate tumor tissues. To test this hypothesis, we used cDNA microarray analysis to determine the transcriptional profiles for 259 transcribed sequences mapping to the 8p chromosomal region for seven human prostate tumor xenografts, completely characterized for numerical and structural alterations on chromosome 8, and five normal human prostate tissues. These experiments identified 33 genes differentially expressed between normal and malignant prostate tissues, the majority of which (28/33, 85%) were transcriptionally down-regulated in malignant compared to normal human prostate tissues. These findings, that haploinsufficiency and transcriptional down regulation for genes mapping to 8p are largely coincident in human prostate tumors, should provide a powerful tool for the identification of tumor-suppressor genes associated with human prostate cancer initiation and progression. PMID- 12759930 TI - Differentiating pathogenic mutations from polymorphic alterations in the splice sites of BRCA1 and BRCA2. AB - About 4% of all BRCA1 and BRCA2 alterations reported to the Breast Information Core database are splice site variants. Only a limited number of them have been studied at the RNA level. By BRCA1 and BRCA2 mutation analysis of breast/ovarian cancer families, we identified two novel and eight previously reported potential splice site mutations, never characterized at the cDNA level before. RT-PCR was performed to determine whether these variants disrupted correct splicing. To ensure efficient detection of transcripts containing premature termination codons, a nonsense-mediated mRNA decay inhibitor was added to the lymphoblastoid cell lines of the patients before RNA extraction. We found that BRCA1 IVS3+3A>C, 4304G>A (in the last codon of exon 12), and IVS19+2delT and BRCA2 IVS6+1G>A, IVS23-2A>G, and IVS24+1G>A lead to aberrant transcripts in lymphocytes. Therefore, they were considered to be true pathogenic mutations, predisposing carriers to cancers of the hereditary breast/ovarian cancer syndrome. BRCA2 IVS24 16T>C is a frequent polymorphism in linkage disequilibrium, with a polymorphic stop codon in exon 27, K3326X. BRCA1 IVS2-14C>T and BRCA2 IVS9-5insT and IVS25+9A>C represent rare variants, not disrupting normal splicing in blood lymphocytes. However, some of the alterations may act differently, qualitatively and/or quantitatively, in breast or ovarian tissues. The data provided in this paper allowed more accurate risk estimation of patients and relatives carrying the mutations described herein and have facilitated genetic counseling. Furthermore, our study is important for a better understanding of splicing mechanisms and revealed new patterns of alternative splicing in BRCA1 and BRCA2. PMID- 12759931 TI - Evidence for whole chromosome 6 loss and duplication of the remaining chromosome in acute lymphoblastic leukemia. AB - HLA class I molecules serve the essential immunological function of presenting antigen to CD8+ T lymphocytes. Tumor cells may present tumor-specific antigen to T cells via these molecules, but many tumors show a loss or down-regulation of HLA class I expression and this may serve as an immune escape mechanism. Using a microsatellite marker-based method, we have searched for loss of heterozygosity (LOH) mutations at 3 genomic regions implicated in HLA class I expression in a cohort of 56 acute lymphoblastic leukemia (ALL) samples. The regions analyzed consisted of the HLA class I heavy chain genes located within the MHC genomic region on chromosome arm 6p, the HLA class I light chain (beta-2-microglobulin, B2M) gene on chromosome arm 15q, and the putative HLA modifier of methylation gene (MEMO1) located on chromosome arm 1q. Results revealed low frequencies of B2M (2/55) and MEMO1 (5/42) LOH but a high frequency of MHC LOH (19/56) that was usually associated with whole chromosome 6 loss (13/19). Cytogenetic data were available for 30 samples, including nine of those that exhibited apparent whole chromosome 6 loss. No cases of chromosome 6 monosomy were observed. We propose that whole chromosome 6 loss with reduplication of the remaining chromosome is common in ALL and that it is driven by the presence of tumor-inhibiting factors on chromosome arm 6p (the HLA loci) along with previously localized tumor suppressor genes on chromosome arm 6q. PMID- 12759932 TI - Insertion of MLL sequences into chromosome band 5q31 results in an MLL-AF5Q31 fusion and is a rare but recurrent abnormality associated with infant leukemia. AB - MLL gene rearrangements leading to production of MLL fusion proteins are commonly detected in infant leukemia patients; the most common MLL fusion associated with infant leukemia is the MLL-AF4 fusion. A single case of chromosomal rearrangement leading to production of an MLL fusion with AF5Q31, a gene structurally similar to AF4, has been detected recently in the malignant cells of an infant leukemia patient. We have identified a second case of MLL-AF5Q31 fusion, arising from an insertion of MLL sequences into chromosome 5, also in an infant leukemia patient. Because MLL and AF5Q31 are transcribed in opposite orientations, a simple balanced chromosomal translocation cannot produce a fusion protein, and complex chromosomal rearrangements such as insertions and inversions are required to produce an MLL-AF5Q31 fusion protein. This report demonstrates that chromosomal insertion of MLL sequences is a rare but recurrent abnormality associated with infant leukemia. PMID- 12759934 TI - Registered nurses' actual and perceived knowledge of diabetes mellitus. AB - This study assessed nurses' actual and perceived knowledge of diabetes. These nurses were employed in a community hospital and home healthcare agency. The Diabetes Basic Knowledge Test (DBKT) and the Diabetes Self-Report Tool (DSRT) were used to assess actual and perceived knowledge. Nurses actual and perceived knowledge were positively correlated (r = 0.402, p < 0.0001). A mean score of 72.2% was obtained on the DBKT. The study findings raise questions about how competent nurses are in caring for patients with diabetes. PMID- 12759935 TI - Using problem-based learning to promote critical thinking in an orientation program for novice nurses. AB - To assist novice nurses in making the transition into the acute care setting, a student-centered, small-group, problem-based program was developed and combined with a preceptorship program. Challenged to work through the problems, novice nurses met the program objectives while enjoying the program. Supervisors and clinical specialists working with the graduates noted increased critical thinking, increased interest and ability in seeking information, and increased problem-solving ability. PMID- 12759936 TI - Cultural nursing care: the planning, development, and implementation of a learning experience. AB - "Cultural diversity" is a nursing education buzz word. Many facilities are adding cultural care competencies and learning experiences to the orientation curriculum. Designing a program to assist nurses to increase their knowledge of cultural issues should be based on a solid theoretical framework and adult learning theories in order to promote a higher level of critical thinking. In this article, the author explains the planning, development, and implementation of such a learning experience. PMID- 12759937 TI - The efficacy of an educational program to improve direct caregiver knowledge regarding fall prevention. AB - One hundred seventy-one (N = 171) direct caregivers were tested to determine their baseline knowledge about patient falls and fall prevention and to determine the impact of a fall prevention education program on their knowledge. Following the pretesting of participants for baseline knowledge a 20-minute educational videotape was viewed. Identical posttesting of knowledge followed. Mean posttest scores (M = 14.01) were significantly higher (t = 17.883; p < 0.001) than the pretest scores, supporting the effectiveness of the educational program in increasing direct caregivers' knowledge related to fall prevention. PMID- 12759938 TI - Effective conflict management begins with knowing your style. AB - The well being of patients depends on the ability of nurses to manage conflict creatively. The Bartol/McSweeney Conflict Management Scale, an assessment tool that helps nurses identify their predisposition toward conflict, is described. The tool is simple to use and suitable for nurses working in different settings. PMID- 12759939 TI - Arranging clinical affiliations. AB - It is frequently the staff educator who is assigned to manage clinical affiliations. It is important to know the regulations that apply to the various professions for which the staff educator is responsible. PMID- 12759940 TI - Educating to manage the accelerated change environment effectively: Part 2. AB - Without appropriate educational preparation, nurse managers may not have the competencies to manage accelerated change effectively. Part 1 of this two-part series provided the condensed findings from an extensive review of nursing, business, and higher education literature. Part 2 describes the results of a Delphi study whereby baccalaureate-prepared nurse manager experts and nurse educator experts in baccalaureate nursing programs validated what linear and nonlinear change management concepts they believed were relevant in managing change in today's dynamic environment. Staff development educators and administrators, as organizational change agents, can use the validated concepts to develop educational offerings to promote effective change management. PMID- 12759941 TI - [Removal of YY1 binding sites in HPV 16 LCR increases viral transforming activities on mouse fibroblasts]. AB - OBJECTIVE: To study the effect of removal of YY1 binding sites within the LCR region of HPV 16 on viral transforming activity. METHODS: Previously we had generated new plasmids carrying HPV 16 whole genome, which contained naturally occurred mutated LCR sequences. The viral transforming abilities on mouse fibroblasts were evaluated in anchorage-independent assays, while the expression and activity of YY1 protein in fibroblasts were tested with EMSA and luciferase assays. RESULTS: YY1 protein was expressed in mouse fibroblasts C127, with ability for DNA binding and repression on P97 activity. Both HPV 16 wild-type DNA and mutated DNAs were transfected into C127 cells and spread to the soft-agarose mediums after selecting with G 418. The growth numbers of the cells transfected with mutated HPV 16 DNAs were 2-10 fold more than that with wild-type HPV 16 DNA. CONCLUSION: Like in epithelial cells or cell lines, transcription regulator YY1 is widely expressed in rodent fibroblasts. Removal of YY1 binding motifs can elevate in the context of the whole genome, the anchorage independent growth ability and the transforming capacity of HPV 16 on the mouse fibroblasts. PMID- 12759942 TI - [Genotyping of human papillomavirus genotyes in cervical scrapes by line probe assay]. AB - OBJECTIVE: To pursue the further development of a practical HPV genotyping system applicable for mass screening. METHODS: Cervical scrapes from 155 women were tested for the presence of HPV using a general-primer based PCR. A reverse hybridization assayor, the line probe assay (LiPA) was used for genotyping 16 different HPV types(HPV 6, 11, 16, 18, 31, 33, 35, 40, 42, 43, 44, 45, 51, 52, 56, 58) simultaneously. RESULTS: 93(60%) out of 155 specimens tested were positive for HPV DNA by PCR. The HPV positivity amongst the group of women with no cervical abnormality was 43%, while that in women with cervical dysplasia and cervical cancer was 81.2% PCR products from 89 HPV positive cases were genotyped by LiPA, of which 49 cases were single HPV infected concerning 11 different HPV types and the commonest type was HPV-16, occurring in 25(51%) of all single HPV infected cases. The other 40 specimens contained multiple HPV types comprising 16 different HPV types. Although double infections(n = 25) prevailed, triple(n = 9), quadruple(n = 3) and quintuple (n = 3) infections were also found. PCR products of four HPV positive cases failed to be classified by LiPA. Sequencing analysis revealed that these 4 cases were HPV-66 which was not included in LiPA probes. Follow-up data of 12 cases confirmed the HPV genotyping results. CONCLUSION: The LiPA for HPV genotyping is easy to perform. It allows accurate and rapid identification of 16 different HPV types in cervical scrapes and will facilitate HPV detection as well as genotyping in cervical cancer screening programs. PMID- 12759943 TI - [Detection of human papillomavirus in laryngeal carcinoma with digoxigenin labelled probe prepared by polymerase chain reaction]. AB - OBJECTIVE: To study the relationship between laryngeal carcinoma and human papillomavirus (HPV), and describe distribution, expression of genome types of HPV in laryngeal carcinoma. METHODS: We used a non-radioactive digoxigenin labeled probe prepared by polymerase chain reaction (PCR) to test the fresh tissue samples from 146 cases with different laryngeal lesions. The labelled probe of HPV consensus primers was obtained, which enabled detection of nine types of HPV DNAs that were HPV 6, 11, 16, 18, 31, 33, 35, 42 and 58. The genome types of HPV DNA positive cases were determined by using multiple primers PCR. RESULTS: The results turned out to be as follows: The HPV infection positive rate was 45.6%(31/68) for laryngeal carcinoma, 20.0% (3/15) for neck metastatic lymphnode, 11.8%(2/17) for precarcinomatous lesions and 6.3%(1/16) for vocal cord polypus. 15 cases of normal tissues adjacent to carcinoma and another 15 normal tissues opposite to the carcinoma were all HPV DNA negative. HPV16, 18 types are the main types of laryngeal carcinoma and HPV6, 11 types often appear in benign lesions of larynx. CONCLUSION: These results implicated that the occurrence and development of laryngeal carcinoma may be closely related to HPV infection. PMID- 12759944 TI - [Cloning and sequencing of human papillomavirus 16 L1 gene from cervical carcinoma tissues of Chinese women]. AB - OBJECTIVE: Human papillomavirus type 16 (HPV16) is highly related with the development of cervical carcinoma. HPV16 late gene L1 encodes its main capsid protein. This study is to analyze the whole sequence of L1 gene of HPV16 of the Chinese isolates. METHODS: Three samples of HPV16 L1 gene were amplified from cervical carcinoma tissues of Chinese patients by PCR and then cloned and sequenced. RESULTS: There were four sites in nucleic acid sequences of all three HPV16 L1 fragments were different from the originally reported sequence of HPV16 and the differed sequences had changed the triplet codes, therefore, subsequently changed the amino acids it coded. CONCLUSION: The results showed that some mutation had taken place in the nucleotide sequence of L1 gene of HPV16 obtained from the cervical carcinoma tissues of Chinese women. PMID- 12759945 TI - [Construction of adenovirus 4 vector with deletion of 78.9-86 mu fragment and express of beta-galactosidase gene]. AB - OBJECTIVE: To construct a human adenovirus type 4(Ad4) vector with partial deletion at E3 region (78.9-86 mu). METHODS: Ad4 DNA was extracted from purified virus cultured in WI-38 cells. The essential fragment(71.3-100 mu) covering Ad4 E3 region was cloned and partial deletion of E3 region of this clone has been performed, generating plasmid pAd4 delta KS. Furthermore, a beta-galactosidase (beta-gal) gene flanked by CMV early promoter and SV40 poly A signal was inserted into pAd4 delta KS, resulting in pAd4c beta. This plasmid was cotransfected with Ad4 DNA, Bcl I A fragments into 293 cells, producing a non-defective recombinant Ad4 virus encoding B-gal. RESULT: The results showed that the constructed recombinant virus could efficiently express the foreign gene, beta-gal. CONCLUSION: Ad4 vector with a deletion of E3 region can be explored as a live vaccine for prevention of human infectious diseases. PMID- 12759946 TI - [The study on cytomegalovirus infection associated with vascular complications in patients with essential hypertension]. AB - OBJECTIVE: This paper studied HCMV infection inpatients with essential hypertension and the relationship of HCMV infection with vascular complications. METHODS: Indirect ELISA method was used to detect human cytomegalovirus (HCMV) specific antibodies in sera of 105 patients with essential hypertension. RESULTS: The results showed that the positive rates of active HCMV-IgM and IgA antibodies were 26.67% and 20.95%, respectively, higher than that of the control group (6.67%). The positive rate of HCMV active IgM antibody in III stage group was significantly higher than that of I + II stage groups (P < 0.01). Active HCMV infective rate was higher in patients with vascular complication. CONCLUSION: These results suggested that HCMV active infection may be associated with vascular complication. PMID- 12759947 TI - [Effect of shen-dong-xin-bao oral liquid on IFN-induction and NK cell activity in CVB3m infected mice]. AB - OBJECTIVE: Our aim was to investigate the effect of Shen-dong-xin-bao oral liquid on IFN-induction and NK cell activity in mice with coxsackie virus B3(CVB3m) infection in different infective stages. METHODS: The level of serum interferon (IFN) and the activity of natural killer (NK) cells in CVB3m induced myocarditis in mice were determined. Natural killer cell cytolytic activity was tested by MTT method and interferon titers by 50% microcytopathic effect assay. RESULTS: In the infected mice fed with 30 g/kg or 12 g/kg per day of Shen-dong-xin-bao oral administration for 10d and 20d, the mean titers of IFN were markedly higher than that in untreated infected group, P < 0.05. The NK cell activity in treated infected groups was also prominently higher than that in untreated infected group, especially on 20d, P < 0.05. CONCLUSION: This results demonstrate that administration of Shen-dong-xin-bao oral liquid possesses enhancement of antiviral immunocompetent action. PMID- 12759948 TI - [Influence of the synthetic peptide recognized by neutralizing antibody on IFN induced biological responses]. AB - OBJECTIVE: The influence of synthetic peptide WLDPRH on IFN-induced biologic activity was studied. METHODS: Based on our earlier studies in which two peptides were recognized by neutralizing antibody from the hexapeptide library displayed on the phage, one of the peptides synthesized, with amino acid sequence WLDPRH. RESULTS: Computer-building approach showed that the synthetic peptide WLDPRH exhibited structural homology with LoopAB (29-35), the binding domain of type I IFN receptor. In the competitive ELISA studies, the synthetic peptide WLDPRH in 25 micrograms/ml could inhibit IFN binding neutralizing antibody 4C1. IFN-induced antiviral assay showed that the protective effects of suboptimal dose (20-70 pg/ml) of IFN were increased from 40% to 86% in the presence of the synthetic peptide WLDPRH and 1.4 micrograms/ml of the synthetic peptide WLDPRH was the lowest dose with the best protection. But potentiating effect on IFN-induced growth inhibition was fewer noticeable in 1.4 micrograms/ml of synthetic peptide. CONCLUSION: The results indicated that synthetic peptide WLDPRH can mimic the structure and activity of Loop AB in the IFN molecule and can be applied to immunotherapy as well as the mimic minimolecular design of IFN. PMID- 12759949 TI - [Cloning the representative strains of China HIV-1 subtypes B, C and E for heteroduplex mobility analysis (HMA)]. AB - OBJECTIVE: To clone the HIV-1 env gene and establish the HMA reagents suitable for use with the Chinese HIV strains. METHODS: Using Nest-PCR technique, we amplified env gene from representative strains which contain the most proximate consensus of China HIV-1 subtypes B, C and E. The genes were then cloned into pGEM-Teasy vector for HMA subtyping. RESULT: The env gene of representative strains of China HIV-1 subtypes B, C and E were amplified and cloned into plasmids. The plasmids were used in HMA and compared with those standard env gene of other countries. The heteroduplex bands obtained with ours env genes were more proximate to homoduplex bands. The homogeneity of the corresponding China HIV-1 subtypes with standard TH14(B), IND868(C), MA959 (C) and TH22(E) are 90.2%(B), 88.3%(C), 83.7(C)% and 92.3%(E) respectively. CONCLUSION: The HMA reagents we established in this study have higher sensitivity in HMA analysis for China HIV-1 strains when compared with the international HMA reagents. The HMA reagents also provide new reference reagents for UNAIDS HIV isolation network. PMID- 12759950 TI - [Sequence analysis of human immunodeficiency virus type 1 tat gene among the long term HIV infected non-progressoris in Yunnan province]. AB - OBJECTIVE: HIV-1 tat gene is one of its regulatory genes necessary for its replication. This study is to explore whether the variation of tat gene influences the AIDS progress of HIV infected individual. METHODS: We carried out investigation on tat gene of HIV among the long term HIV infected non-progressors and the commonly infected individuals. Mononuclear cells of peripheral blood were isolated and cellular DNA was extracted. Nested PCR method was used to amplify tat gene and their sequences were analyzed. RESULTS: Blood were collected from 22 HIV infected individuals, of which 11 cases were infected with B subtype strains based on previous env gene analysis and belonged to long term non-progressors according to clinical features and immunology. The other 11 cases were common HIV infection of which 5 subtype B and 6 subtype C were found. Both long term non progressor group (subtype B) and the commonly HIV infectedly group (subtype B) had constantly four amino acids different from that of international B subtype consensus sequence The both groups clustered randomly on the phylogenetic tree. In commonly HIV infected subtype C group there were 6 amino acids different from that of international subtype C. The variation inside the tatgene of common subtype C group was small. CONCLUSIONS: There was no regular variation and significant difference between long term HIV infected non-progressive group and commonly HIV infected subtype B group. There is no evidence showing that progressive speed of AIDS correlated with the tat sequence profile of HIV-1. PMID- 12759951 TI - [The prognostic value of virology and immunology markers in Chinese HIV-1 infected people]. AB - OBJECTIVE: To study the prognostic value of virology and immunology markers in Chinese HIV-1 infected people and their correlation. METHODS: We studied 190 HIV infected people/AIDS patients' samples from China. Plasma viral load, P24 antigen, virus culture, CD4 positive T lymphocyte (CD4) count and CD8 positive T lymphocyte(CD8) count were tested. Test groups were divided according to clinical staging diagnosis from physicians. The data were analyzed in aspects of their relations with clinical status and correlations among all the above markers. RESULTS: Plasma viral load, CD4 count and CD4/CD8 ratio showed significant difference between asymptomatic stage and symptomatic stages (P < 0.01), also between asymptomatic and AIDS stages(P < 0.001). Virus culture positive rate increased significantly from asymptomatic to symptomatic stage. No significant clinical change was observed associating with P24 antigen positive rate. CD8 count decreased slightly when AIDS occurred (P < 0.001). Close correlation was observed between plasma viral load, CD4 count and CD4/CD8 ratio. CONCLUSION: Viral load, CD4 count and CD4/CD8 ratio are useful indicators to AIDS prognosis and virus culture can be a supplementary marker. p24 antigen detection seems no diagnostic value to disease progress. The significance of CD8 change will be further studied. PMID- 12759952 TI - [Analyze T lymphocyte subsets of HIV/AIDS patients by flowcytometer]. AB - OBJECTIVE: Using flowcytometer (FCM) to detect CD4+, CD8+ lymphocytes in peripheral blood, combine with clinical symptoms to evaluate the HIV/AIDS patients' immune state. METHODS: Anti-coagulated peripheral blood were obtained from 8 HIV-AIDS patients and 5 normal persons. After analyzing their total white cell counts, the whole blood was stained with two-color immunofluorescence using directly conjugated monoclonal antibody pairs, followed by lysis of red erythrocytes, fixation of lymphocytes, and analysis by flowcytometry. Multiplying the CD4 and CD8 percentages by the absolute number of lymphocytes obtained from the total white blood cell (WBC) count divide by the lymphocyte differential percentage, we got the CD4, CD8 counts. RESULTS: The HIV/AIDS patients' CD4 counts were lower than those of normal controls'. Especially, the AIDS patients' CD4 counts were all below 200 cells/mm3 and their clinical symptoms were serious. CONCLUSION: The HIV/AIDS patients' CD4 counts are highly correlated with their clinical symptoms. The result also shows that FCM is accurate, sensitive and reliable in CD4 count. PMID- 12759953 TI - [Bio-safety testing for retroviral vector as gene therapy delivery system]. AB - OBJECTIVE: To generate a bio-safety testing system including the testing of sterility, mycoplasma and replication-competent retroviruses(RCR) in the study of gene therapy anti-hepatitis B virus by intracellular immunization. METHODS: Mycoplasma was detected by polymerase chain reaction. S+/L- assay, NIH3T3 amplification and rescue assay of neo gene were performed to determine RCR. RESULTS: The results showed that all of the packaging cell lines were negative for aerobic, anaerobic bacteria and fungi. One of the packaging cell lines was positive for mycoplasma. No RCR was detected in all of the packaging cell lines. CONCLUSIONS: The results suggested that the methods using in present report are stable and sensitive, it is very useful for bio-safety testing of gene therapy in clinical trials. PMID- 12759955 TI - [Detection of viral markers in liver tissue dually infected with hepatitis B and C viruses by double labelling]. AB - OBJECTIVE: To study the distribution and mutual relation of HBV and HCV in dually infected liver tissue. METHODS: Detected HBV and HCV nucleic acids and their antigens by immunohistochemistry and in situ hybridization, the cases with HBcAg and HCV RNA positive were tested further on single section by double labeling. RESULTS: On the section tested by double labeling we could see that positive hepatocytes infected with HBV or HCV were mostly present in different liver lobules, even if the two viruses were seen in the same lobule, most of them were not seen within a single cell. CONCLUSIONS: In most of hepatocytes, there was no direct interference or suppression of each other in the liver tissue dually infected with HBV and HCV. Double labeling method is useful to analyze the coinfection of two viruses in the same tissue. PMID- 12759954 TI - [HBsAg expression, anti-HBs induction and pathological observation in the mice inoculated with DNA vaccine against hepatitis B]. AB - OBJECTIVE: To study if special humoral immunological response can be induced by DNA vaccine against Hepatitis B (NV-HB/s) and its mechanism. METHODS: The Balb/C mice inoculated with NV-HB/s by intramuscular injection were detected for HBsAg(the expressed product of NV-HB/s) by ABC immunohistochemistry or ELISA, and for anti-HBs by ELISA and also performed the routine pathological examination. RESULTS: After inoculated with NV-HB/s, HBsAg could be detected from mice muscle tissue samples of injected sites one week later and almost kept positive(75% and even more) up to 6th month, but was undetectable from all the serum samples during this period; anti-HBs could be detected from serum samples of some mice two weeks later and of all the mice detected one month later, it still kept positive at 6th month while the mice were sacrificed. Under microscope, only non specific inflammation was found in the muscle tissues of injected sites and could completely recover within 4 weeks, no matter whether the mice were inoculated with NV-HBs, or with traditional HB vaccine derived from blood, or even with PBS as control. CONCLUSION: DNA vaccine against Hepatitis B (NV-HB/s) can successfully express HBsAg in the muscle tissue of mice by intramuscular inoculation and subsequently induce anti-HBs in vitro. PMID- 12759956 TI - [Dynamic changes of basal core promoter and pre-core mutants of hepatitis B virus in patients with chronic hepatitis B virus infection]. AB - OBJECTIVES: To clarify the association of hepatitis B virus mutants in basal core gene promoter and pre-c region with serum HBeAg state and persistent HBV infection. METHODS: 32 patients, who infected with hepatitis B virus and suffered from several exacerbation during the course of this disease, were followed up for an average of 20.8 months. Using two mis-matched primers to amplify BCP and pre-c gene fragments, in combination with restricted fragment length polymorphism assay, we studied the presence of these two mutants(nt 1762A-->T, 1764G-->A and nt1896G-->A) in 105 serum samples. Direct sequencing was performed on 15 patients' serial samples to identify the efficiency of these rapid and simple methods and other variations in these regions. RESULTS: The presence of these two kinds of mutants increased during the following-up, (62.5% > 46.9%; 31.3% > 12.5%). In most cases, two mutants prevailed and finally displaced the wild type virus; the serology of HBeAg was affected predominantly by the ratio of BCP mutant to wild type. CONCLUSION: BCP and pre-c mutants emerged in the course of chronic infection and were selected positively by the inflammation activity. The serological conditions of HBeAg were changed when the predominant situations of these two mutants were established. The survival of these two mutants implied that mutations in these two regions may contribute to the persistent infection of hepatitis B virus. PMID- 12759957 TI - [Significance of polypeptide antigen of HCVE2/NS1 relative conservation region in detection of anti-HCV]. AB - OBJECTIVE: In order to study the significance of HCVE2/NS1 relative conservation region antigen. METHODS: EIA was established with HCVE2/NS1 relative conservation polypeptide antigen, as well as serum HCV RNA and ALT in patients with HCV infection. RESULTS: The positive rate of 96 cases patients who were infected by HCV is 41.67%, acute 13.55% and chronic 25.04%, and we found that there were 3 cases weak positive in 40 healths; Agreement rate is 62.5% in 40 cases of the positive of anti-E2/NS1 in sera with HCVRNA, and they have relation. In ALT level in sera, acute infection of HCV is higher than chronic (P < 0.05). CONCLUSION: There are virus necleic acid in sera when anti-HCVE2/NS1 appearence positive; however, antibody of anti-E2/NS1 relative conservation region has the some neutralive actions, and this humor immunity may have the actions in clearing virus, but wether anti-body of anti-E2/NS1 relative conservation of HCV is or not neutralive antibody well be investigated further. PMID- 12759958 TI - [Study on HCV genotypes in different clinical types of hepatitis C patients in Shenyang area]. AB - OBJECTIVE: To study the distribution of HCV genotypes in different clinical types of hepatitis C in Shenyang area and its clinical significance. METHODS: The HCV genotypes of 84 HC patients of different clinical types were detected by PCR assay with four kinds of type-specific primers. RESULTS: In 84 patients with hepatitis C, the HCV-II and HCV-III genotypes and the II/III mixed genotype were 53.6%, 30.9% and 15.5% respectively. The distribution of HCV genotypes in different HC clinical types was not identical, P < 0.05. In acute hepatitis, in mild, moderate and severe degree of chronic hepatitis, and in liver cirrhosis, the infection rates of genotype II HCV were 55.6%, 36.2%, 75.0%, 85.7% and 88.9% respectively, but that of genotype III were 22.2%, 46.8%, 8.3%, 0% and 11.1% respectively. CONCLUSION: Infection of geno type II HCV was predominant in Shenyang area followed by HCV-III and II/III mixed genotypes. The different genotypes of HCV were related to the severity of hepatitis C. PMID- 12759959 TI - [Detection of hepatitis C core antigen C33]. AB - OBJECTIVE: To investigate the distribution of C33 antigen of HCV in serum and liver tissues from normal subjects and patients with different diseases. METHODS: Balb/C mice were immunized by HCV core antigen peptide C33(33 amino acids) which was conjugated with mice serum albzmin by a heterotypic difunctionalconnector(SPDP). Two strains of monoclonal antibody to C33 were produced. By using one of them as coated and the other as HRP(horseradish peroxidase)-labelled antibodies, an ELISA was developed to detect C33 antigen of HCV in 1236 serum samples from patients. RESULTS: The detectable rate of peptide C33 was 14.0%(6/43) in acute hepatitis C group, 8.8%(35/396) in anti-HCV-IgG positive group, 6.5%(4/62) in chronic hepatitis C group, 4.3%(1/23) in leukemia group, 3.3%(19/576) in hemodialysis group, 1.5%(2/136) in other diseases group, and 1. 4%(6/438) in blood donors. Results of immunohistochemistry and immunofluorescence for liver tissues obtained from biopsy showed that C33 peptide was found in nucli, cytoplasm and membranes of hepatic cells, scattered or laterally concentrated in the cytoplasm. CONCLUSION: The detection of C33 antigen of HCV may consider as one of the markers of HCV infection. PMID- 12759960 TI - [Dynamic observation of serological and histological changes of Chinese rhesus monkeys infected by hepatitis G virus]. AB - OBJECTIVE: To study the genesis, development and pathogenesis of hepatitis G. METHODS: The Chinese Rhesus monkeys were intravenously injected with the serum from a hepatitis G patient only with HGV RNA positive. The dynamic changes of serum and liver tissues of the animals were observed before and after infection. Also the immunohistochemical study were done with monocolonal antibody against NS5 gene antigen of HGV. One male monkey was dissected and the internal organs were taken for histological examination 18 months after infection. RESULTS: Serum ALT and AST became increasing 30 days after infection and showed abnormal continuously for 9 months. Spotty and focal necrosis and piecemeal necrosis were found in liver tissue from 2 to 18 months after infection. Positive staining of HGV antigen was present in cytoplasm of liver cells by immunohistochemistry. Histological observation showed that the internal organs were normal except the liver tissue in which the damages were similar with that in hepatitis. CONCLUSION: The successful infection of Chinese Rhesus monkey by serum from hepatitis G patient can used as a good animal model for the study of hepatitis G. PMID- 12759961 TI - [Preliminary study on NS5 region gene sequencing and variants of hepatitis G virus in China]. AB - OBJECTIVE: To study the genomic specificity and variation of Hepatitis G virus(HGV) in China. METHODS: The 141nt cDNA fragments locating at nonstructural region 5 (NS5) of HGV were obtained from the sera of 16 HGV infected patients by RT-nested-PCR using NS5 primers, then purified and directly sequenced by dideoxynucleotide chain termination method. RESULTS: The sequence diversity near the N-terminus of the region between nt 7,820 and 7,960 was more marked than that near the C-terminus. Compared with the HGV-US isolate, the nt alterations of NS5 in Chinese isolates usually took place at nt 7,824, 7,892, 7,914, 7,917, 7,935 and in region nt 7,845-7,860, the rate of variability at one nt position was 0 89.5%. The nucleotide homology was 83.6%-95.7% (89.6% in average) and the homology of deduced amino acid sequence was 83.0-100%(91.2% in average). CONCLUSION: The nucleotide and deduced amino acid sequences of NS5 of Chinese HGV are relatively conserved but are obviously different from the HGV and GBV-C reported in USA in this study. PMID- 12759962 TI - [Detection of IgM-antibody to hepatitis G virus and its clinical significance]. AB - OBJECTIVE: To provide a rapid and early serodiagnostic technique for the patients with hepatitis G virus(HGV). METHODS: A capture ELISA was developed to detect anti-HGV IgM in sera using synthetic peptides of HGV NS3 NS5 gene regions conjugated with horseradish peroxidase. RESULTS: This assay was not disturbed by competition of specific IgG or interfered by RF and did not cross-react with HAV, HBV, HCV, HEV, CMV and EBV. Among 46 sera of non-A-E hepatitis, 14 were positive for anti-HGVIgM, the positive rate was 30.43%, 6 was positive for HGV RNA the positive rate was 42.86%. 12 paired sera from acute stage of HGV patients were all positive by capture-ELISA. CONCLUSION: This method is sensitive, specific, rapid and stable for detecting anti-HGV IgM, and is useful in the early diagnosis of HGV infection. PMID- 12759963 TI - [The pathogenicity of HFRS patient's soft palate mucosa to suckling mice]. AB - OBJECTIVE: To explore the pathogenic mechanism of hemorrhagic fever with renal syndrome(HFRS). METHODS: 2-3 days old inbred Balb/C suckling mice were inoculated intracerebrally with tissue suspension of soft palate petechia of HFRS patients and the HFRS virus RNA was tested in brain, lung and kidney tissues of sucking mice by nested PCR. RESULTS: Some of the mice developed acute disease and died. The autopsy analysis indicated that these mice showed pathological changes in microvascular system and parenchymatous tissues. HFRS virus RNA was detected in their brains, lungs and kidney tissues. CONCLUSIONS: These results indicate that HFRS virus exists in the tissue of petechial hemorrhagic spots on soft palate petechia of HFRS patients. PMID- 12759964 TI - [Nucleotide sequence of A/Goose/Guangdong/2/96 (H5N1) virus M and NS RNA]. AB - OBJECTIVE: To determine the nucleotide sequences of M and NS genes of A/Goose/Guangdong/2/96(H5N1) virus and also to compare them with the sequences of A/HK/156/97(H5N1) strain for revealing the relationship between the two viruses, as well as for setting up a solid base for studying M and NS genes of influenza A viruses in the future. METHODS: Virion RNA was transcribed into cDNA by reverse transcriptase, cDNA was amplified by PCR, the products of PCR were purified. Afterward, RNA sequence analysis was performed by the dideoxynucleotide chain termination method using synthetic oligodeoxynucleotide primers. RESULTS: The segment length of A/Goose/Guangdong/2/96(H5N1) virus RNA 7 is 1,027 nucleotides. It codes M1 (252 amino acids) and M2 (97 amino acids) proteins. However, the segment length of RNA 8 of A/Goose/Guangdong/2/96(H5N1) virus is 890 nucleotides coding NS1 (230 amino acids) and NS2 (121 amino acids) proteins. The amino acid sequence homologies of M1, M2, NS1 and NS2 protein molecules between A/Goose/Guangdong/2/9(H5N1) and A/HK/156/97 (H5N1) viruses are 97.6%, 92.8%, 65.7% and 76.9%, respectively. CONCLUSION: The lengths of A/Goose/Guangdong/2/96(H5N1) virus RNA 7 and RNA 8 and 1,027 are 890 nucleotides, respectively. These two RNA segments belong to those of avian influenza A virus. The A/HK/156/97(H5N1) virus RNA 7 and RNA 8 segments are not derived from those of A/Goose/Guangdong/2/96(H5N1) virus directly. PMID- 12759965 TI - [Establishment of a method for classification of HBV genome and it's application]. AB - OBJECTIVE: By using the techniques of PCR and restriction fragment length polymorphism (RFLP), we established a classification method for HBV genome based on viral S gene of asymptomatic HBV carriers (AsC). METHODS: By using this method we classified HBV DNA of AsC living in Guangzhou, Shenyang, Beijing and Chongqing. RESULTS: Among the AsC of Guangzhou, genotype B was 32.8%, genotype C 42.7%, mixed genotypes B and C 23.0%, others 1.6%; among the AsC of Chongqing, genotype B was 35.0%, genotype C 40.0%, mixed genotypes B and C 25.0%; among the AsC of Beijing, genotype B was 25.0%, genotype C 50.0%, mixed genotypes B and C 25.0%; among the AsC of Shenyang, genotype B was 11.1%, genotype C 88.9%. The prevalent HBV strains in China are genotype C and genotype B, and genotype C is the main genotype in west China. CONCLUSION: The established method for genotyping is simple and convenient. Using this method, we also classified HBV DNA of the patients with chronic hepatitis B and liver cirrhosis living in Guangzhou, and found that mixed genotypes B and C was in 50.0% of the patients detected, suggesting the mixed infection might lead to a severe damage of the liver tissue. PMID- 12759967 TI - [Expression and characterization of two outer capsid proteins VP2 and VP5 of bluetongue virus in insect cells]. AB - OBJECTIVE: To study the biological characteristics of the outer capsid proteins VP2 and VP5 of bluetongue virus (BTV) expressed in insect cells and their potential use in the assembly of BTV and genetic engineering vaccine. METHODS: The genes which encode the two outer capsid proteins VP2 and VP5 of bluetongue virus (BTV) 10 were separately cloned into pFastBac1 vector and the corresponding recombinant baculoviruses were obtained. RESULTS: BTV VP2 could be expressed in Sf-9 cells better than VP5. Further works indicated that VP2 could elicit neutralizing antibodies to BTV10(1:64), and also could partially neutralize BTV1(1:16), but could not neutralize BTV13. The co-immunizing of VP2 and VP5 could induce higher neutralizing antibodies to BTV10 and BTV1. VP2 also showed a hemagglutination activity. CONCLUSION: VP2 expressed in insect cells could induce neutralizing antibodies to BTV and had the biological activity of hemagglutination, VP5 could enhance the ability of neutralizing antibody induction of VP2, they can be used for the assembly of virus-like particles and for the development of genetic engineering vaccine. PMID- 12759966 TI - [Complete nucleotide sequence of the nonstructural gene of alphavirus YN87448 strain isolated in China and its relationship to other Sindbis viruses]. AB - OBJECTIVE: To determine the complete nucleotide sequence of the nonstructural gene of YN87448 virus stain which was firstly isolated from a female patient with fever in Yunnan Province in 1986, and identified as a member of Alphavirus by the serological method. METHODS: The complete nucleotide sequence of the nonstructural region gene of YN87448 virus strain was determined with nine clones, which were obtained by using reverse transcription and polymerase chain reaction (RT-PCR), and by linking nine overlapping fragments into pGEM-T vector respectively. RESULTS: The complete nucleotide sequence of nonstructural gene of YN87448 virus strain was 7,613 nucleotides long exclusive of the 5' cap, encoding four nonstructural proteins, nsP1, nsP2, nsP3, nsP4, and contained one initiator(ATG) and two stop codons (TGA). In comparison with the consensus sequence of S.A.AR86, the homogeneity of the nucleotide sequence between YN87448 virus strain and sindbis-like virus isolate S.A.AR86 was 98.8%. YN87448 virus strain can not produce a fatal disease in adult mice. In comparison with the consensus sequence of S.A.AR86, there is a 54 nucleotide insertion from 5,256 bp to 5,309 bp, 3 nucleotide (AGT) deletion at 5,603 bp in nsP3 region, and an opal termination codon between the nsP3 and the nsP4 genes in YN87448. virus strain. This sequence has been put into Gene Bank and No. is AF103734. CONCLUSION: YN87448 is a new sindbis-like virus strain. PMID- 12759969 TI - [A preliminary clinical safety evaluation of parvovirus H-1 on cancer therapy]. AB - OBJECTIVE: To make a preliminary evaluation of parvovirus H-1 for its safety in clinical use. METHODS: In this study, clinical safety of H-1 in cancer therapy was examined using the Ames test, the mammalian cell chromosome aberration test, the micronucleus calculation of mouse bone marrow cells, the allergy test and the pyrogenicity test. RESULTS: Negative results were observed in all the tests. The mouse tolerance test showed that the tolerance limit was at least 5 x 10(11) plaque-forming units/kg weight, which was 2500 times higher than the supposed clinical dose. The H-1 host NB-324K cells were also tested using the anchorage independent assay and the tumour formation in nude mice. Both of the assays gave negative results. CONCLUSION: All these results preliminarily showed that the H-1 was safe in cancer therapy. PMID- 12759968 TI - [No switch region is found in the regulatory sequence of HPV 16 for promoter P97]. AB - OBJECTIVE: Cellular transcriptional regulatory factor YY1 represses the activity of the early promoter P97 of HPV 16, but the regulatory activity to early promoter P105 of HPV 18 depends on a switch region which locates at the upstream sequences of YY1 binding sites. This study is to find out whether within the HPV 16 LCR there is such motif. METHODS: DNA sequences of the whole HPV 16 LCR region were analyzed. Two similar switch structures were found. Different CAT reporter plasmids, one containing HPV 16 reference and the other mutated LCRs, both starting at the end of L1 and from the beginning of the enhancer, were generated and transfected in HeLa cells transiently. RESULTS: CAT assays showed there is no change of regulation pattern of YY1 on P97, when the two switch-similar structures were deleted. Sequences analysis also showed that no switch region like motif could be demonstrated in enhancer region, in promoter proximal segment, and even in E6/E7 area. CONCLUSION: It suggests that there is no such switch region within HPV 16 transcriptional regulatory region. PMID- 12759970 TI - [Detection of TT virus DNA in peripheral blood mononuclear cells in patients with chronic hepatitis B and hepatocellular carcinoma using PCR and in situ hybridization]. AB - OBJECTIVE: To determine whether the TTV infection occurs in peripheral blood mononuclear cells (PBMCs) of patients with chronic hepatitis B (of medium degree) and in HBsAg positive hepatocellular carcinoma (HCC). METHODS: TTV DNA in PBMCs was detected by nested-polymerase chain reaction(PCR) and in situ hybridization (ISH). RESULTS: 7 of 26 cases of chronic hepatitis B were positive for TTV DNA by nested-PCR with a positive rate of 26.9% that was significantly higher than that of normal control(chi 2 = 14.3, P < 0.001); 4 of 21 HBsAg positive cases were positive for TTV DNA by nested-PCR with a positive rate of 19.0%, also significantly higher than that of normal control(chi 2 = 4.86, P < 0.05). 4 of 7 TTV positive patients with chronic hepatitis B tested by nested-PCR were positive for TTV DNA by ISH in PBMC cytoplasm, with a positive rate of 57.1%. CONCLUSION: (1) A higher incidence of TTV DNA in PBMCs of patients with chronic hepatitis B and in HBsAg positive HCC was identified by nested-PCR and ISH; (2) TTV DNA was in the cytoplasm of PBMCs in patients with chronic hepatitis B. PMID- 12759972 TI - [Characterization of HA1 genes of influenza A (H1N1) viruses isolated in Shenzhen City]. AB - OBJECTIVE: To understand the molecular bases of intention of influenza virus activity and emergence of "O" phase of influenza A(H1N1) strains in human population in Shenzhen in recently years, and also the evolutionary characterization of influenza A(H1N1) HA1 gene. METHODS: Virion RNA was transcribed into cDNA by reverse transcriptase, cDNA was amplified by PCR, the products of PCR were purified. Afterward, RNA sequence analysis was performed by the dideoxynucleotide chain termination method using synthetic oligodeoxynucleotide primers. Finally, phylogenetic analysis of the sequencing data was performed with MegAlign (version 1.03) and Editseq (version 3.69) softwares. RESULTS: Since 1995, there were three different genetic lineages of influenza A(H1N1) virus HA1 gene cocirculating in men in Shenzhen city. Adding one and deleting one of potential glycosylation sites at 54 and 155 positions of amino acid sequences on HA1 protein domain of H1N1 viruses isolated recently was found as compared with those of A/Singapore/6/86(H1N1) virus. Meanwhile, there were some differences of amino acid sequences on HA1 protein molecules among H1N1 viruses tested and A/Singapore/6/86 (H1N1) virus. CONCLUSION: The intention of influenza A(H1N1) virus activity occurred since 1995 was due to emergence of substitution in amino acid sequences, especially the appearance of one addition and one deletion of potential glycosylation sites on their HA1 protein domains, and also was due to occurrence of influenza A(H1N1) virus with feature of "O" phase. PMID- 12759971 TI - [Relationship between HBV DNA serum level and acute exacerbation of the disease in chronic hepatitis B patients]. AB - OBJECTIVE: In this study, we evaluated the relationship between HBV DNA serum level and acute hepatic exacerbation of the disease in chronic hepatitis B patients. METHODS: A series of serum samples from 14 patients of acute exacerbation of chronic hepatitis B were analyzed for alteratieus in serum HBV DNA and alanine transaminase (ALT) level before, during, and after episode of an acute exacerbation. The serum HBV DNA concentrations were tested by the quantitative polymerase chain reaction (PCR) using the AmpliSensor assay. RESULTS: 1. Serum HBV DNA average level in 11 patients significantly increased from 7.148 +/- 2.008(logarithm) at 2-8 weeks before maximum injury of liver to 8.416 +/- 2.160 (logarithm), the time that ALT reached its peak, and then, following the declining of ALT level, decreased to 6.093 +/- 1.428 (logarithm). In 9 of 11 cases, the HBV DNA peak value occurred before the ALT peak level or at the same time; 2. Two patients who experienced several episodes of acute exacerbation showed that the appearance of acute hepatic injury and the relatively normal liver function occurred alternatively. Whereas, one patient with liver cirrhosis showed persistence of high level viremia in three episodes of acute exacerbation in one year. CONCLUSION: 1. This investigation revealed a clear correlation between increasing level of serum HBV DNA and acute exacerbation of patients with chronic hepatitis B. The findings suggest that host's immune response which causes acute exacerbation of liver injury in patients with chronic hepatitis B is triggered by the change of viremia and HBV replication; 2. Patients with severe chronic active hepatitis and persistent hepatitis B virus replication are at very high risk of rapid progression to cirrhosis. PMID- 12759973 TI - [The influence of interferon alpha neutralizing antibodies on the therapeutic effectiveness in patients with chronic viral hepatitis]. AB - OBJECTIVE: To study the influence of interferon alpha (IFN-alpha) neutralizing antibody (NA) on the therapeutic effectiveness of IFN-alpha. METHODS: 30 healthy persons and 116 patients with chronic viral hepatitis treated with IFN-alpha were studied, and the NA in their sera were tested by using the antiviral neutralizing bioassay (ANB). RESULTS: None was positive for NA in sera of the healthy persons and patients before IFN-alpha treatment, 20 of 116 patients developed NA after IFN-alpha therapy, the positivity was 17.2% (20/116). NA appeared after 2 months of alpha IFN treatment, all 20 patients were positive after 6 months. The NA positivity in treated groups of IFN-alpha 2a, IFN-alpha 2b and IFN-alpha 1b were 34.6%, 13.2% and 11.5% respectively. The clearance rates of virus in NA-positive group and NA high-titered group were significantly lower than those in NA negative group and NA low-titered group (P < 0.01 and P < 0.05, respectively) In contrast, the relapse rates of virus in NA-positive group was markedly higher than that in NA-negative group (P < 0.05). CONCLUSION: NA may develop with its regularity in the chronic viral hepatitis patients treated with alpha IFN. The frequencies of NA development are related with the subtypes of alpha IFN used. NA can inhibit IFN activity, thereby affecting the therapeutic effectiveness of IFN. PMID- 12759974 TI - [Optimization of productivity of Epstein-Barr virus membrane antigen in baculovirus infected insect cells]. AB - OBJECTIVE: To probe factors influencing the expression level of in insect cells. METHODS: Insect cells at different growth phases and in different culture media were infected with recombinant baculovirus carrying EBV-MA gene, where the membrane anchor sequence was deleted. The expression level of EBV-MA was detected by immunodots assay and scanner analysis. RESULTS: Cells at the early exponential phase yielded more expressed recombinant EBV-MA than those at the late exponential and stationary phases. Infected cells supplemented with fresh medium at various phases of growth resulted in improved productivity of EBV-MA. Further experiments revealed that replacing fresh medium prior to infection and supplementing a mixture of glucose and glutamine after infection, the productivity could be improved dramatically. Comparing the productivity of EBV-MA expressed in sf9 cells with that in sf21 cells, no obvious difference was found. CONCLUSION: Consumption of nutrients and accumulation of metabolites in cell culture media were the important factors influencing the productivity of EBV-MA in insect cells. PMID- 12759975 TI - [Detection of human papillomavirus gene in biopsies from colon carcinoma by PCR]. AB - OBJECTIVE: To detect the human papillomavirus gene (HPVs) in biopsies from colon carcinoma by PCR and to study the relationship between HPV infection and colon carcinoma. METHODS: 46 biopsies from colon carcinoma, 10 biopsies from tissue around carcinoma and 16 control biopsies were collected and DNA were extracted. The HPV DNA in biopsies were amplified by PCR using general primers (GP) and primers for HPV6/11, HPV16/33 and HPV18. RESULTS: 20 out of 46 biopsies from colon carcinoma showed HPV gene positive, HPV6/11, HPV6/33 and HPV18 positivity were 3, 11 and 6 respectively. HPV DNA was found in 1 case from 10 biopsies of surrounding tissue of carcinoma. 16 control biopsies were negative. CONCLUSION: HPV may be related to the etiology of colon carcinoma. PMID- 12759976 TI - [Preliminary study on relationship between different viral pathogenesis and disease prognosis in patients with severe viral hepatitis]. AB - OBJECTIVE: To study the relationship between different viral pathogenesis and disease prognosis in severe viral hepatitis. METHODS: Different viral pathogenesis of 87 dead and live cases with severe viral hepatitis were compared. RESULTS: Total mortality of 87 patients with severe hepatitis was 74.71% (65/87), total prevalence of HBV infection alone in these patients was 41.38% (36/87). The detection rates of HBV infection alone and superinfection of different hepatitis viruses in 68 patients with chronic severe hepatitis (CSH) were 41.18% (28/68) and 58.82% (40/68) respectively. The prevalences of superinfection of HBV and HEV or HAV and superinfection of HBV and CMV in patients with CSH were 27.94% (19/68) and 10.29 (7/68) respectively. The mortality of superinfection of HBV and CMV (85.71%) was the highest, followed by HBV infection alone (77.78%). In addition, the prevalence and mortality of HBV infection alone in 19 patients with acute or subacute severe hepatitis was the highest. CONCLUSION: HBV, HEV or HAV infection alone was the main viral pathogenesis of severe hepatitis. Superinfection of different viruses in patients with CSH was the most common viral infection type. An unpromising prognosis of superinfection of HBV and CMV in CSH is noted. PMID- 12759978 TI - [In situ enzyme immunoassay for detection of hepatitis A virus]. AB - OBJECTIVE: To establish a more simple and sensitive technique for identification of hepatitis A virus (HAV). METHODS: HAV vaccine strain H2M20-K and wild-type strain He34 were used to infect KMB17 deploid cells in 96-well tissue culture plates and in situ enzyme immunoassay was used to detect viral infection with specific HA monoclonal antibody, A(OD) value of 1.8 was used as positive indicator. RESULTS: Observation of viral replication showed the peak titer of viral infectivity reached at 20-25 days after viral infection, which was 4-6 day searlier than that of ELISA endpoint assay. Comparing the viral infectivity titers of 13 individual samples, the two methods revealed no significant statistic difference (t = 1.13, P > 0.05). Our tests showed that in situ EIA could better reflect the results of neutralization and 5 HAV positive isolates were directly isolated from 12 faecal samples after the first passage culture. CONCLUSION: Although the sensitivities of the two tests were similar, the direct detection of virus/cell system with in situ EIA showed simple, easy to operate and reproducible, it may replace the conventionally used ELISA endpoint assay for identification of HAV, even for the other viruses. PMID- 12759977 TI - [Efficient expression of human papillomavirus type 16 (HPV16) capsid proteins L1 and L2 in E. coli]. AB - OBJECTIVE: To study the expression of HPV16 capsid proteins L1 and L2 in E. coli. METHODS: The plasmids pET-30a-L1 and pET-30a-L2 were constructed from the pET-30a vector, and transducted into the BL21. The fusion proteins 6 x His-L1 and 6 x His L2 were expressed when induced by adding of IPTG. SDS-PAGE and Western blot analyses were used to detect the expressed proteins. RESULTS: Fusion proteins 6 x His-L1 and 6 x His-L2 were expressed efficiently in E. coli, the Mr were about 60,000 and 97,000 respectively, 6 x His-L1 degraded more than 6 x His-L2. CONCLUSION: L1 and L2 proteins were expressed at a high level in prokaryotic expression system, and 12 protein were more stable than L1 protein. PMID- 12759979 TI - [Renal impairment after spontaneous bacterial peritonitis in chronic severe viral hepatitis]. AB - OBJECTIVE: To study the clinical features, prognosis and influence factors of renal impairment (RI) in chronic severe viral hepatitis with spontaneous bacterial peritonitis (SBP). METHODS: 129 discharged patients with chronic severe viral hepatitis(CSVH) complicated with spontaneous bacterial peritonitis were analyzed retrospectively. RESULTS: Renal impairment is a frequent event in CSVH patients with SBP. SBP-R I is functional in nature, it may be followed by a rapidly progressive or a stable disease course, or may be reversed after recovery of the infection. CONCLUSION: Independent prognostic factors were age, clinical course of liver disease, other infection or diabetes, severe renal impairment, serum sodium level before SBP and negative response to antibiotic therapy. PMID- 12759980 TI - [Analysis of genome character and amino acid residues 2,209 to 2,248 of NS5 A region of hepatitis C virus in relation to the response to interferon therapy]. AB - OBJECTIVE: To observe nucleotide sequence variations and amino acid residues 2,209 to 2,248 of the NS5 A region of hepatitis C virus in relation to interferon therapy in Chinese patients infected with HCV genotype 1b. METHODS: We collected sera from 22 patients infected with HCV genotype 1b, performed nested RT-PCR using extracted serum RNA from sera, amplified NS5 A region 2,209 to 2,248 and sequenced the PCR product directly. The deduced amino acid sequences were compared with the corresponding region of the HCV-J prototype strain. RESULTS: Only 4 isolates were intermediate type, and no mutant type was found. In 18 patients treated with IFN-alpha, only 4 were HCV RNA negative at present, No correlation was found between amino acid mutation and IFN response. CONCLUSION: Variations in the NS5 A region between amino acid residues 2,209 to 2,248 failed to predict IFN response in Chinese patients infected with HCV genotype 1b. IFN may accelerate nucleotide variation but has little influence on amino acid variations. PMID- 12759981 TI - [The detection of virus antigen in the lower respiratory tract of the patients with lung cancer]. AB - OBJECTIVE: To find out about the viral infection situation of lower respiratory tract of the patients with lung cancer. METHODS: The excretion from the surface of bronchiogenic carcinoma was brushed under fibrobronchoscopy. The respiratory virus antigen was detected and analysed by reagent kit produced by the 262nd Hospital of Beijing Military Region. RESULTS: The respiratory virus antigen was positive in eight cases of lung cancer group, the positive rate was 17.4%(8/46), it was significantly higher than that in non-lung cancer group (P < 0.05). Among them, there were one case of influenza virus A, two cases of influenza virus B, two cases of para-influenza 1,3, two cases of adenovirus and one case of respiratory syncytial virus. The carcinoma accompanied with viral infection were 4,3,1, cases in order of squamous carcinoma, small cell lung carcinoma and adenocarcinoma. CONCLUSION: The results showed that a relationship existed between lung cancer and viral infection of respiratory tract statistically. The viral infection increased in patients with lung cancer, this is worthy to pay attention to. PMID- 12759982 TI - [A mutation specific polymerase chain reaction for detecting hepatitis B virus genome with A-to-C mutation at nt585]. AB - OBJECTIVE: To investigate the prevalence of hepatitis B virus (HBV) nt585 A-to-C mutants in China. METHODS: A mutation specific polymerase chain reaction (msPCR) was established for amplifying HBV DNAs with A-to-C mutation at nt585. Two sets of primer pairs with same sequences except for one base at 3' terminus were designed and synthesized according to 48 of the known HBV genome sequences and the popular HBV subtypes, adr and adw, in China. RESULTS: 57 of HBV S gene fragments amplified by a nested PCR from different and child adult hepatitis B patients were characterized by using the msPCR. It was shown that HBV mutants with A-to-C mutation at nt585 could be confirmed. CONCLUSION: The msPCR is a specific and sensitive method for detecting the HBV nt585 A-to-C mutants. PMID- 12759983 TI - [Detection of Epstein-Barr virus gene in nasopharyngeal carcinoma with neck lymphatic metastasis]. AB - OBJECTIVE: To investigate the presence of Epstein-Barr virus(EBV) gene in neck lymphatic metastasis of nasopharyngeal carcinoma. METHODS: Fifty patients' masses in the neck were punctured and the fluids were analyzed with PCR for EBV DNA. RESULTS: Twenty seven cases showed EBV DNA positive among thirty cases of nasopharyngeal carcinoma with neck lymphatic metastasis, and the positive rate was 90%. One case was EBV DNA positive in seven cases of malignant lymphoma and the positive rate was 14.3%. One case was EBV DNA positive in eight tumor patients without NPC metastasis and the positive rate was 12.5%. Five cases with nasopharyngeal inflammation and neck lymphatic node proliferation were demonstrated to be EBV DNA negative. CONCLUSION: This technique is rapid, sensitive and specific, which is valuable clinically, especially for the diagnosis in patients with cryptic nasopharyngeal carcinoma and with neck metastasis but lack of clear original focus. PMID- 12759984 TI - [Molecular epidemiology of respiratory syncytial virus infection in infants]. AB - OBJECTIVE: To search for the molecular epidemiology of respiratory syncytial virus (RSV) infection in infants. METHODS: The randomly amplified polymorphism DNA (RAPD) technology was used to study 96 RSV strains isolated and identified from the patients in Changchun State Hospital of Pediatrics during 1992-1994 and 1 reference strain of M type. RESULTS: All strains were typable, four RAPD patterns were observed, different diseases have different types. 87.14% of the patients' RSV strains had the same RAPD pattern(R1). CONCLUSION: RAPD method is a rapid, easy and high resolution technique, with which we can know on the molecular level the microbial infection state. There was epidemic of R1 type infection during the period surveyed. PMID- 12759985 TI - 37 trillion hits!! Now what?! The benefits of literature searching. PMID- 12759986 TI - Amniotic fluid embolus: a review of the research literature. AB - The diagnosis of Amniotic Fluid Embolus (AFE) is often missed clinically leading to the incorrect attribution of a major post-partum haemorrhage, which may accompany AFE, to uterine atony. This paper reviews the research on AFE with particular emphasis on the clinical presentation and diagnosis; both before and after death. It begins by dealing with the difficulty of defining AFE due to confusion about the underlying pathophysiological events. As this paper will demonstrate, the theory that AFE is an embolic event is no longer valid. A description of the clinical manifestations is provided so that these can be explained by the contemporary theory of AFE as an 'anaphylactoid' reaction. Finally, the difficulties of diagnosis, particularly laboratory diagnosis, will be discussed. The research indicates that it is not possible to accurately diagnose AFE, either, pre or post mortem, by any currently available laboratory tests. Because of better diagnosis it is now known that AFE in not uncommon and, if it is diagnosed early, a much higher rate of intact survival can be achieved than was previously thought possible. PMID- 12759987 TI - A survey of midwives who participated in a randomised trial of perineal massage in labour. AB - A multi-centred randomised perineal massage in labour trial (PMLT) was conducted in which participating midwives randomised eligible women in the second stage of labour. A survey of these midwives was conducted after completion of the PMLT, but before results had been analysed and presented. The aim of the study was to seek from midwives, following the PMLT, their reasons why some eligible women were not randomly allocated to a group; why others did not receive care as allocated and the midwives' views about the massage, including whether significant trial results would influence their clinical practice. (This paper presents the results of this survey.) PMID- 12759989 TI - Desert Storks--Bush midwives. PMID- 12759988 TI - Experience-based educational strategies to promote woman-centred midwifery practice. AB - This paper discusses the evaluation of educational activities developed and implemented in a tertiary based, pre-endorsement midwifery program in Brisbane, Australia. The development of the experience-based activities outlined was driven by the feminist philosophy of the curriculum and a commitment to helping students develop as woman-centred practitioners. The activities focused on enabling students to hear women's stories of childbearing firsthand and understand these stories from the women's perspective, thus validating personal experience. PMID- 12759990 TI - The role of staff development in assuring competence. AB - Leaders and educators from many facilities are struggling to understand what is expected of them, relative to competence assessment, by the joint Commission on Accreditation of Healthcare Organizations (JCAHO). Some have responded to the standards with the creation of a large number of checklists and annual skills fairs, along with a fragmented competence assurance system--all delegated to the staff development department. This article discusses such approaches, reviews JCAHO requirements, and suggests alternative strategies, including the use of the Nursing Interventions Classification. PMID- 12759991 TI - Service and education share responsibility for nurses' value development. AB - This article examines professional values of senior baccalaureate nursing students and practicing nurses. An important finding was that practicing nurses rated behaviors reflecting values in the American Nurses Association (ANA) Code for Nurses as more important than did senior students, thereby supporting the notion that practice contributes to value formation. The ongoing development and internalization of the nursing professions' values requires active involvement by staff development educators. The phenomena of value formation and development of professional values appear to mirror the novice to expert model. PMID- 12759992 TI - Competency program development across a merged healthcare network. AB - Competency validation from initial entry through continued employment is essential to ensure nursing personnel provide safe professional practice. With hospital mergers it becomes quickly apparent that varying methodologies for competency verification exist. Nursing staff educators need a system-wide competency program to ensure a single high standard for the entire healthcare network. This article addresses one healthcare network's response to the challenge of developing and implementing a standardized competency-based program across a wide variety of settings. PMID- 12759993 TI - The caregiver's careshop. A renewal experience for nursing staff. AB - What are some creative methodologies that staff development educators can use to nurture nursing staff while promoting caring and compassionate behaviors? The authors describe an innovative process used during a 1-day workshop designed to convey caring to nursing staff through a variety of experiences. The overall goal of the session is to provide caregivers with a variety of new "tools" to care for themselves as they deal with multiple stressors in their personal and professional lives. PMID- 12759995 TI - Guidelines for improving resident dining experiences in long-term care facilities. AB - This article is a part of a larger study that identified nutrition education needs for nurses and nursing assistants that resulted in an educational intervention. The purpose of this article is to introduce newly developed evidence-based dining room guidelines. These guidelines can be used as a part of nursing homes' quality improvement programs to evaluate dining services and assist facility staff to identify problem areas. PMID- 12759994 TI - A review of nurses' performance of cardiopulmonary resuscitation at cardiac arrests. AB - This study investigated nurses' ability to initiate and maintain effective cardiopulmonary resuscitation (CPR) in actual cardiac arrests (n = 50). Results indicated that in the majority of cases nurses effectively managed all components of CPR. These results are in direct contrast to the current accepted body of knowledge related to deficits in nurses' CPR performance. In particular the results suggest that nurses' actual management of cardiac arrests in a contextual environment differs markedly from results shown by research using simulated settings. Research findings suggest the need for an evaluation of the use and effects of cognitive and metacognitive instructional strategies in CPR training courses on transfer of skills and knowledge to practice. PMID- 12759996 TI - New service in a community hospital: the role of nursing staff development. AB - The purpose of this article is to describe strategies to assist nurses in staff development with planning and implementing a successful education program directed at preparing staff to provide a new patient service in a community hospital. The staff development instructor can play a crucial role in the successful implementation of a new service in a community hospital. This article focuses on the implementation of a cardiac surgery program in a community hospital. PMID- 12759997 TI - Professional nursing burnout and irrational thinking. AB - This article reports how professional, job-related burnout in nurses (N = 192) is examined in relation to a developed index of irrational thinking patterns in a large, urban hospital setting. Based on the constructs of Rational Emotive Behavior Therapy (REBT), the study examines maladaptive thinking patterns related to nursing burnout and provides insight into possible educational and staff interventions for the syndrome. Low mean scores on all but two subscales indicate overall strength and stability among this sample. The demonstration that both burnout thoughts (r = 0.451, p = < .01) and burnout behaviors (r = 0.350, p = < .01) are significantly correlated with the perfection and control pattern support the study's assumptions. Nurses who demand perfection and control in themselves and others create unrealistic demands and expectations that cannot be met in the real world of nursing. The investigator believes that a regular stress management program, using the concepts of REBT, can foster professional growth and development, decrease workplace conflict and stress, and provide nurses (and other employees) with strategies and tools to disarm the irrational beliefs that build maladaptive cognitive patterns leading to professional burnout. PMID- 12759998 TI - A patient-centered approach to nurse orientation. PMID- 12759999 TI - Making the transition to a hospital-wide department of education: strategies for success. AB - In today's healthcare environment, staff development specialists have experienced a shift from a nursing-focused approach to education to an organization-wide approach. The change fosters both challenges and opportunities for the staff development specialist. This article describes the strategies used by a department of nursing education to successfully transition to a hospital-wide department of education. PMID- 12760000 TI - Using the hospital Intranet to meet competency standards for nurses. AB - Regulatory agencies increasingly require hospitals to provide evidence of employee compliance with mandated competencies. This article provides information on the innovative use of Intranet-delivered computer-based training as an effective and efficient method of providing and documenting training to meet regulatory requirements. The authors discuss the advantages and disadvantages of Intranet-based instructional modules, instruction on developing Intranet-based modules, and an example of a healthcare system that is effectively using intranet based modules for competency training. PMID- 12760001 TI - Using criticisms to elicit and meet unit-based learning needs. AB - A strategy using staff nurses' criticisms is described, which may be a useful addition to the repertoire of strategies used by staff development educators to elicit and meet unit-based learning needs. It is also argued that this strategy has the potential to equip staff nurses to move more independently from criticism to problem setting and problem solving. PMID- 12760002 TI - A continuous curriculum for building code blue competency. AB - Staff response to a code blue can be cultivated when a continuous curriculum for code blue competency is used. This curriculum details ongoing code blue education yet requires only small increments of time for the inservice classes. The curriculum, adaptable to any unit, consists of three components: a unit-specific orientation to emergency equipment; exercises in critical thinking and doing; and exercises in documentation. PMID- 12760003 TI - Enhancing the learning in self-directed learning modules. AB - Self-directed learning modules are a popular option for providing continuing nursing education. Not all nurses are comfortable with this learning format. Nursing educators may be unfamiliar with strategies to facilitate self-directed learning. Self-directed learning is based on assumptions about adults as learners. Effectiveness and acceptance of the learning modules are enhanced by educational strategies that address these assumptions. PMID- 12760004 TI - Use of multiple teaching strategies in the staff development setting. AB - The purpose of this study was to determine if using a variety of teaching strategies in the staff development setting would affect learner outcomes with regard to knowledge acquisition and participant satisfaction. A control group class was presented with the use of lecture only. An experimental group class was presented using a variety of teaching strategies. Posttest scores from the experimental group were significantly higher than those from the control group, as were satisfaction levels. Findings indicate that the use of a variety of teaching strategies can result in positive learner outcomes. PMID- 12760005 TI - The ethics of midwifery. PMID- 12760006 TI - Policy or tradition: oral intake in labour. AB - Oral intake restrictions have varied over time and around the world with written hospital policies for this issue often being non-existent. As there are differing views on this issue within hospitals overseas, a survey was conducted of 109 maternity units in New South Wales, Australia during early 2000 to identify the trends across the state. In New South Wales 81.7% of hospitals did not have a written policy for oral intake in labour. The remaining 18.3% had written policies which varied in their oral intake allowances from ice only to whatever women feel like eating and drinking. Of the 109 hospitals in this survey 60.5% leave food and fluid requirements to the individual woman's discretion, providing they have no increased risk of general anaesthetic. PMID- 12760007 TI - Antenatal education classes in Victoria: what the women said. AB - Thirteen women's experiences in antenatal education classes emerged as significant during the data analysis of a much larger study on women's experiences of their first pregnancy. Two grounded theory procedures, the making of comparisons and the asking of questions were used to analyse the antenatal classes experiences. Subcategories Unprepared (labour and birth experiences), and Vulnerable (breastfeeding) evolved during the grouping of the concepts. Tape recorded, individual interviews were held with the women in their homes for about one hour on four occasions. The classes were completed in the 3rd trimester; this discussion, therefore, relates to the third trimester and post birth interviews only. Ten women were delivered in private hospitals and three women in public hospitals. Comments about the classes during the third interview were generally positive. Following birth, comments were less favourable. Audit trials were created for the larger study of which the present topic was a small part. PMID- 12760008 TI - The partograph. Used daily but rarely questioned. AB - The partograph is part of the midwifery tools of practice, used everyday, but rarely questioned. This paper explores the history of the partograph and describes its development from 1954. It examines the components of the partograph and discusses when one should be commenced. The value of the partograph in practice today is questioned. PMID- 12760009 TI - Home too soon? A comment on the early discharge of women from hospital after childbirth. AB - The current trend of health care services to encourage early discharge of women from hospital within three days of childbirth has produced some concerning issues. This paper explores the factors surrounding this trend and highlights the need for a postpartum midwifery community infrastructure to support women who are discharged from hospital before breastfeeding and parent education processes are established. PMID- 12760011 TI - Reasons for increased postnatal length of stay & women's perceptions of length of stay. AB - A quality improvement project was conducted by a major maternity provider (3000 births per year) in the western suburbs of Melbourne. The first part of the project explored the reasons for increased length of stay (LOS), beyond the average timeframe for women receiving postnatal care. The perceptions of 100 women on their length of stay was also measured in the second part of the project. Two information audits were conducted to explore reasons for increased LOS. These reasons were varied, and by the time of the second audit, there was a demonstrated reduction in LOS. A Length of Stay Questionnaire was used to investigate women's perceptions of their LOS on discharge from hospital, and by telephone follow-up 14 days post-discharge. Perceptions varied between women and related to issues such as individual health complications, models of care, postnatal education, and degree of support at home. This project has implications for postnatal midwifery care in relation to discharge planning and women's expectations of care following birth. PMID- 12760010 TI - Management of second stage labour with an epidural--a problem in isolation? AB - This paper examines practices in second stage labour of a woman with an epidural. Common practices are compared with the current literature. Recommendations from this comparison are that practices such as directed pushing should be discouraged and pushing should be delayed until the presenting part has descended to a vaginal station greater than +1. Implications for practice are discussed and recommendations made to implement changes. PMID- 12760012 TI - The role of the midwife when the pathology report states your pregnant client has bacterial vaginosis. AB - Bacterial vaginosis (BV) has been associated with preterm labour, preterm prelabour rupture of membranes and preterm birth. Organisms in high concentrations in BV may ascend from the vagina into the uterine cavity causing infection. Many factors may change the vaginal pH and microflora increasing the risk of BV, and midwives need to recognise signs and symptoms of BV. The vaginal pH is a useful tool for BV screening. Midwives may play a future role in pH testing, especially in pregnant women at high risk, for example, those with a previous preterm birth. PMID- 12760013 TI - Pramwalking as postnatal exercise and support: an evaluation of the Stroll Your Way To Well-Being program and supporting resources in terms of individual participation rates and community group formation. AB - Stroll Your Way To Well Being is a community pramwalking program designed to increase access by mothers to sociable postnatal exercise. This study evaluated the program in terms of long-term adherence rates by mothers to walking, and the effectiveness of kits distributed to providers in assisting them to set up their own groups. Approximately one quarter of information kit recipients established a pramwalking group. Reasons for non-initiation included lack of time or a perceived lack of priority. Mothers reported program benefits to be the opportunity to exercise, socialize and share information about baby issues. 70% of mothers were still walking 16 months after the program's commencement. Reasons for mothers ceasing participation in pramwalking included lack of interest, no group leader available or their baby was too old. Future studies need to focus on the prevention and treatment of postnatal depression through exercise. PMID- 12760014 TI - Students and independent midwifery practice in Australia. PMID- 12760015 TI - Understanding the complexity of an organism's responses to DNA damage. PMID- 12760016 TI - DNA repair systems and bacterial evolution. PMID- 12760018 TI - The biochemical basis and in vivo regulation of SOS-induced mutagenesis promoted by Escherichia coli DNA polymerase V (UmuD'2C). PMID- 12760017 TI - Adaptive mutation in Escherichia coli. PMID- 12760019 TI - Human DNA polymerase kappa: a novel DNA polymerase of unknown biological function encoded by the DINB1 gene. PMID- 12760020 TI - Role of yeast and human DNA polymerase eta in error-free replication of damaged DNA. PMID- 12760021 TI - Roles of DNA polymerase zeta and Rev1 protein in eukaryotic mutagenesis and translesion replication. PMID- 12760022 TI - Xeroderma pigmentosum variant: from a human genetic disorder to a novel DNA polymerase. PMID- 12760023 TI - Streisinger revisited: DNA synthesis errors mediated by substrate misalignments. PMID- 12760024 TI - Roles of Watson-Crick and minor groove hydrogen bonds in DNA replication. PMID- 12760025 TI - Structural and mechanistic studies on repair of 8-oxoguanine in mammalian cells. PMID- 12760026 TI - Envisioning the fourth dimension of the genetic code: the structural biology of macromolecular recognition and conformational switching in DNA repair. PMID- 12760027 TI - Repair of endogenous DNA damage. PMID- 12760029 TI - DNA polymerase beta and mammalian base excision repair. PMID- 12760028 TI - Base removers and strand scissors: different strategies employed in base excision and strand incision at modified base residues in DNA. PMID- 12760030 TI - Photolyase/cryptochrome family blue-light photoreceptors use light energy to repair DNA or set the circadian clock. PMID- 12760031 TI - DNA damage recognition and nucleotide excision repair in mammalian cells. PMID- 12760032 TI - Regulation of nucleotide excision repair in bacteria and mammalian cells. PMID- 12760033 TI - Three-dimensional structure and function of replication protein A. PMID- 12760034 TI - Transcription-coupled repair of oxidative DNA damage in human cells: mechanisms and consequences. PMID- 12760036 TI - DNA mismatch repair: from structure to mechanism. PMID- 12760035 TI - Signaling mismatch repair: the mechanics of an adenosine-nucleotide molecular switch. PMID- 12760037 TI - New recognition mode for a TG mismatch: the atomic structure of a very short patch repair endonuclease-DNA complex. PMID- 12760038 TI - Temporary and permanent mutators lacking the mismatch repair system: the enhancement of mutators in cell populations. PMID- 12760039 TI - Molecular manifestations and molecular determinants of telomere capping. AB - Multiple interacting components of the telomere, together with telomerase (and sometimes recombination), determine whether a telomere will be functional, allowing cell proliferation. The various components reinforce each other, providing for a robust and resilient system of protection and replenishment of telomeres. A characteristic of a telomere is that its structural features elicit responses that allow it to be dynamically recapped. Eliciting a DNA damage response through uncapping of a telomere appears to be one way in which telomerase action at that telomere is stimulated. Thus, as long as a timely and appropriate recapping of the telomere is possible, regulated uncapping of a telomere appears to be not only normal, but even required for optimal telomere maintenance. Telomere length and the presence of telomerase provide an example of a pair of interacting components that determine telomere capping function. Telomerase is dispensable in cells with sufficiently long telomeres; but in cells with short telomeres lacking telomerase, cells lose the ability to proliferate, and in some cell types, telomere fusions are increased. However, expressing telomerase can make even very short telomeres functional. Many interesting questions remain as to the mechanisms of these biological effects. PMID- 12760040 TI - A new connection at human telomeres: association of the Mre11 complex with TRF2. PMID- 12760041 TI - Telomere length, telomere-binding proteins, and DNA damage signaling. PMID- 12760042 TI - All things must end: telomere dynamics in yeast. PMID- 12760043 TI - Sir2: an NAD-dependent histone deacetylase that connects chromatin silencing, metabolism, and aging. PMID- 12760044 TI - Arrest, adaptation, and recovery following a chromosome double-strand break in Saccharomyces cerevisiae. PMID- 12760045 TI - Double-strand break repair in human cells. PMID- 12760046 TI - Controlling S-phase onset in fission yeast. PMID- 12760047 TI - Coping with and recovering from hydroxyurea-induced replication fork arrest in budding yeast. PMID- 12760048 TI - Damage response and dNTP regulation: the interaction between ribonucleotide reductase and its inhibitor, Sml1. PMID- 12760049 TI - Is Rad9p upstream or downstream from Mec1p? PMID- 12760050 TI - DNA damage checkpoint control of mitosis in fission yeast. PMID- 12760051 TI - Mitotic checkpoints, genetic instability, and cancer. PMID- 12760052 TI - Microscopy-induced radiation damage, microtubules, and progression through the terminal stage of G2 (prophase) in vertebrate somatic cells. PMID- 12760053 TI - Complexity of damage produced by ionizing radiation. PMID- 12760054 TI - Complex transcriptional responses to macromolecular damaging agents: regulatory responses specific for SN2 alkylation and the MAG1 gene. PMID- 12760055 TI - The interplay between nonhomologous end-joining and cell cycle checkpoint factors in development, genomic stability, and tumorigenesis. PMID- 12760056 TI - S-phase functions of the Mre11 complex. PMID- 12760057 TI - The FHA domain, a phosphoamino acid binding domain involved in the DNA damage response pathway. PMID- 12760058 TI - The FHA domain in DNA repair and checkpoint signaling. PMID- 12760059 TI - Details and concerns regarding the G2/M DNA damage checkpoint in budding yeast. PMID- 12760060 TI - Establishment of and recovery from damage checkpoint requires sequential interactions of Crb2 with protein kinases Rad3, Chk1, and Cdc2. PMID- 12760061 TI - Analysis of the fission yeast checkpoint Rad proteins. PMID- 12760062 TI - A conserved role for the Hus1 checkpoint protein in eukaryotic genome maintenance. PMID- 12760063 TI - DNA-damage-induced checkpoint pathways in the nematode Caenorhabditis elegans. PMID- 12760064 TI - The transcriptional program following p53 activation. PMID- 12760065 TI - Regulation of p53 after different forms of stress and at different cell cycle stages. PMID- 12760066 TI - The DNA damage checkpoint and human cancer. PMID- 12760067 TI - DNA damage responses and chemosensitivity in the E mu-myc mouse lymphoma model. PMID- 12760068 TI - Genetic instability, oncogenes, and the p53 pathway. PMID- 12760069 TI - Multiple signaling pathways involving ATM. PMID- 12760070 TI - ATM: sounding the double-strand break alarm. PMID- 12760072 TI - The function of BRCA1 in DNA damage response. PMID- 12760073 TI - Recombination between two chromosomes: implications for genomic integrity in mammalian cells. PMID- 12760071 TI - Interactions of the Nijmegen breakage syndrome protein with ATM and BRCA1. PMID- 12760074 TI - Transcription-coupled repair: a multifunctional signaling pathway. PMID- 12760075 TI - Chromosomal instability and cancer predisposition: insights from studies on the breast cancer susceptibility gene, BRCA2. PMID- 12760076 TI - Roles of RecQ family helicases in the maintenance of genome stability. PMID- 12760077 TI - Role of DNA break-sensing molecule poly(ADP-ribose) polymerase (PARP) in cellular function and radiation toxicity. PMID- 12760078 TI - Biological responses to DNA damage: a perspective in the new millennium. PMID- 12760079 TI - A step-by-step approach to the evaluation of clinical studies with therapeutic or preventive objectives. PMID- 12760080 TI - Understanding the components of publication success: a survey of academic award recipients. AB - PURPOSE: This study examines predictors of publication number in career development awardees. We examined whether daily writing predicted publication number among junior faculty. DESIGN AND METHODS: We surveyed 94 career development awardees; the survey consisted of 28 questions in four domains: characteristics, environment, writing practices, and attitudes about writing. RESULTS: Variables that contributed positively to publication number included male gender and those with a negative effect were clinical research and perceiving the need to write as a requirement for advancement. In subgroup analysis of junior faculty, a habit of writing daily was predictive of greater publication numbers. IMPLICATIONS: Career development awardees published more first-authored manuscripts if they were male, were involved in nonclinical research, and did not perceive writing as a requirement for advancement. These factors highlight the need to explore the lower overall publication productivity in women and in clinical investigators. Junior faculty members that write daily publish more manuscripts, regardless of gender, research type, or motivators. The benefits of daily writing warrant direct study if not empiric implementation. PMID- 12760081 TI - Economic credentialing: are hospital privileges contingent upon skills--or economics? PMID- 12760082 TI - War and medicine. PMID- 12760083 TI - The Connecticut Nursing Home Quality Initiative--Part II. PMID- 12760084 TI - Experiences in challenging chiropractic. PMID- 12760085 TI - Physicians in "the war against medicine". PMID- 12760086 TI - Having it your way. Three physicians who were early adopters and who remain vocal EMR advocates share their insights about the utilization, challenges and benefits of EMRs. PMID- 12760087 TI - Smart storage. Texas cancer center ensures continuous treatment by using emerging storage networking technology. PMID- 12760089 TI - What works Awards 2002. Boston Medical Center. PMID- 12760090 TI - What Works Awards 2002. Capitol ENT. PMID- 12760091 TI - What works Awards 2002. Horizon Radiology. PMID- 12760092 TI - Biometric identification. Ultrasonic systems can succeed where optical systems may not. PMID- 12760093 TI - What works. Payer-direct claims. For a Pennsylvania health system, the key to claims efficiency was a combination of technology and a do-it-yourself attitude. PMID- 12760094 TI - What works. All-around efficiency. Massachusetts OB/GYN practice improves its financial and administrative health with practice management software. PMID- 12760095 TI - Change--for better or worse? PMID- 12760096 TI - No complaints, big savings from e-commerce venture. PMID- 12760097 TI - Price survey. Suture price hikes slow to a crawl. PMID- 12760099 TI - Sharing the wealth in western New York. PMID- 12760100 TI - [Drug therapy of rheumatoid arthritis]. PMID- 12760101 TI - [Reduced expression of TCR zeta chain in T cells of patients with systemic lupus erythematosus]. PMID- 12760102 TI - [Diversity of virus-infected cells in EB-virus related diseases]. PMID- 12760103 TI - [Epidemiological study on patients with systemic sclerosis receiving financial aid for treatment]. AB - We conducted an epidemiological study of systemic sclerosis in Japan using the records of patients who had been registered to receive public financial aid. A total of 10,956 patients were registered as having systemic sclerosis in 1999. We statistically analyzed the data on the patients, including sex, age, major symptoms, and laboratory findings. We also made contingency tables in order to evaluate the correlations of the data. Our study estimated that the male/female ratio was 1:7.3. The mean ages of male and female patients were 58.8 and 58.5 years old, respectively. The major symptoms were as follows: Raynaud's phenomenon 92.4%, skin sclerosis 94.7%, dyspnea 29.9%, and dysphagia 32.2%. With respect to specific antibodies, antinuclear antibody was present in 92.2%, antitopoisomerase I antibody in 27.5%, anticentromere antibody in 37.7%, and antiribonucleoprotein antibody in 19.9%. Making contingency tables, we could elucidate the association of antitopoisomerase I with lung fibrosis. PMID- 12760104 TI - [The comparison in double immunodiffusion and western blotting methods of anti-SS A/B antibodies in patients with Sjogren's syndrome]. AB - OBJECTIVE: To investigate the autoimmune responses against SS-A/B antigens by double immunodiffusion (DID) and western blotting (WB) in primary and secondary Sjogren's syndrome (SS). PATIENTS: Forty-nine patients with primary SS (PSS), 28 patients with secondary SS (SSS) and control group that couldn't be diagnosed as SS were included in this study. RESULTS: In DID analysis, Anti-SS-A antibody was detected in 69% of PSS and 86% of SSS, and anti-SS-B antibody was found in 22% of PSS and 39% of SSS. No significant difference could be demonstrated between PSS and SSS concerning anti-SS-A/B antibodies. Conversely, WB studies disclosed evidences that 18% of PSS and no SSS reacted only with the 52 kD protein, and there was significantly increased in PSS. Sera reacting with the 60 kD antigen were found in 37% of PSS, 71% of SSS, and 75% of SSS with SLE, 63% of SSS with RA. The ratio of SSS, and SSS with SLE were particularly significantly higher than PSS. CONCLUSION: Our results revealed data that there are the difference of reactivity against SS-A/B antigens in WB between PSS and SSS. PMID- 12760105 TI - [Severe type of Epstein-Barr virus associated hemophagocytic syndrome successfully treated with T-COP-E and splenectomy]. AB - A 16-year-old girl was admitted to our hospital because of high fever, abdominal pain, and jaundice. Abnormal lymphocytes and hemophagocytic cells had infiltrated the bone marrow. Laboratory data revealed a severe type of hemophagocytic syndrome accompanied by an initial Epstein-Barr virus (EBV) infection. Persistent EBV infection was identified by polymerase chain reaction (PCR) detection of EBV DNA in peripheral blood and bone marrow mononuclear cells. The limited efficacy of initial treatment with high-dose gamma-globulin, plasmapheresis, and high-dose methylprednisolone prompted us to administration of T-COP-E (VP-16). Two courses of T-COP-E improved the patient's clinical symptoms and laboratory data; however, marked splenomegaly remained. In addition, fever and serum increase of lactate dehydrogenase (LDH) and cytokines such as gamma-interferon recurred shortly after chemotherapy. On day 53 after diagnosis, the patient underwent laparoscopic splenectomy. The resected spleen weighted 420 g and abnormal lymphocytes in the spleen were positive for CD 8 and negative for CD 56. In situ hybridization revealed EBV-encoded small RNAs (EBERs) in the abnormal lymphocytes. Clinical symptoms including high fever disappeared shortly after the splenectomy, and laboratory data returned to normal. Lymphocytosis after the splenectomy was not observed. We continued out patient monitoring of the case, and 16 months after diagnosis, EBV-DNA in peripheral blood mononuclear cells was not detected, even by PCR. PMID- 12760106 TI - The risks of germline gene transfer. PMID- 12760107 TI - The last transfer. PMID- 12760108 TI - Medical marijuana. PMID- 12760109 TI - Smallpox vaccine: not worth the risk. PMID- 12760110 TI - Of mice and men: patenting the oncomouse. PMID- 12760111 TI - Realignment of research priorities in the FY 2004 budget. PMID- 12760112 TI - The judicial dismantling of the Americans with Disabilities Act. PMID- 12760113 TI - Removing the mask. PMID- 12760114 TI - Facing Carrie Buck. PMID- 12760116 TI - Too much of a good thing: how splendid technologies can go wrong. PMID- 12760117 TI - Accident & desire. Inadvertent germline effects in clinical research. PMID- 12760118 TI - Inheritable genetic modification and a brave new world. Did Huxley have it wrong? PMID- 12760120 TI - African American nurse educators and health disparities. PMID- 12760119 TI - Lethal language, lethal decisions. PMID- 12760121 TI - African American caregivers and HIV/AIDS formal resources. AB - The African American community continues to be disproportionately affected by the human immunodeficiency virus/acquired immune deficiency syndrome (HIV/AIDS) in many ways. In addition to the prevalence rates of the disease in the community, caregivers of people living with HIV/AIDS continue to be challenged as they strive to provide care and support to love ones. The purpose of this pilot study is to explore how African American caregivers of adults living with HIV/AIDS engage available formal resources within their community. African American caregivers (N = 10) were recruited from Long Island, New York and interviewed using unstructured and semi-structured questions. The caregivers' use of community resources were associated with circulating communication within the community about the resource. For example, formal (health & social) resources were recommended and/or accessed if the resource had a positive, historical past of rendering culturally sensitive and medically competent care to the African American community. Recommendations for further study and community- level implications for health care professionals are suggested. PMID- 12760122 TI - Promoting the scholarship of research for faculty and students. AB - Research has become a highly valued activity for nurses. Students at the undergraduate and graduate levels are experiencing more and more research in their programs. As part of the faculty role in higher education, promotion and tenure are tied to the scholarship of research. Yet many schools of nursing do not have a well-developed capacity of nursing faculty members who have the knowledge and skills to competently engage in a research program of their own or to guide and inspire students to pursue a research career. This paper is an attempt to challenge administrators to more aggressively promote research, and faculty and students to think more intensively about the scholarship of research. The challenges of developing a program of research for promoting faculty and students' research are discussed. Strategies for promoting faculty research and advice for beginning researchers are included. PMID- 12760123 TI - Nursing accreditation: what's a librarian got to do with it? AB - Nursing education accreditation visits are similar to JCAHO accreditation visits. Both types of accreditation are offered to ensure quality programs and services. The author, a librarian, describes the role of the nursing library in helping the Hampton University School of Nursing to achieve full accreditation from the two national nursing accreditation bodies. This article describes the process and outcomes of accreditation visits. PMID- 12760125 TI - Casebook: diarrhoea. PMID- 12760124 TI - Key developments in gastroenterology. PMID- 12760126 TI - Caring for patients with colostomies. PMID- 12760127 TI - The patient with stomach cancer. PMID- 12760128 TI - Managing diverticular disease. PMID- 12760129 TI - The management of dyspepsia and reflux. PMID- 12760130 TI - Alcohol-related liver disease. PMID- 12760131 TI - The GP's role in allergic rhinitis. PMID- 12760134 TI - Nursing service. Interview by Tom Keighley. PMID- 12760135 TI - Moving house or moving jobs: what's the difference? PMID- 12760136 TI - Nursing. A European perspective. PMID- 12760137 TI - Role models. PMID- 12760138 TI - Lessons from LEO. PMID- 12760139 TI - The modern matron. PMID- 12760140 TI - Protecting your family. PMID- 12760141 TI - Budgeting: Part Two. PMID- 12760142 TI - Mental unblock. PMID- 12760143 TI - Give and take. Why every black person needs to know about organ donation. PMID- 12760144 TI - The HSJ interview: Professor Peter Hutton. Alternative medicine. Interview by Alastair McLellan. PMID- 12760145 TI - Primary care. Outside chance. PMID- 12760146 TI - Engaging clinicians. Skeleton staff. PMID- 12760147 TI - E-novation. Spam with everything. PMID- 12760148 TI - E-novation. Frontline London. PMID- 12760149 TI - E-novation. Spies like us. PMID- 12760150 TI - E-novation. The telly vision. PMID- 12760152 TI - Limit staff's contact with possible SARS patients. PMID- 12760153 TI - APCs: lack of knowledge can hurt your bottom line. PMID- 12760151 TI - Will your ED have staff quarantined for SARS? Brace yourself for the worst. AB - One Albuquerque, NM, ED's staff were quarantined after exposure to a suspected severe acute respiratory syndrome (SARS) patient, which underscores the importance of reducing risks of transmission. Reduce the number of contacts by isolating patients immediately. Triage nurses should have quick access to N-95 respirators. Ask security to screen all patients arriving by car before they enter the ED. PMID- 12760154 TI - Know these facts before facing smallpox vaccine. AB - Participants in the smallpox vaccination program report minimal disruption to staffing and very few adverse outcomes. There have been no cases of transmission from health care workers to patients. Consider giving staff members an excused day off if they feel ill after receiving the vaccine. Vaccinated staff must perform daily bandage checks and appropriate hand hygiene. PMID- 12760155 TI - Accreditation update. Continuous compliance: starting in 2006, your ED must be prepared for unannounced accreditation surveys. PMID- 12760156 TI - Accreditation update. Quality, timely care: an ED's keys to compliance. PMID- 12760158 TI - Mammographic computer-aided detection systems. AB - While mammography is regarded as the best means available to screen for breast cancer, reading mammograms is a tedious, error-prone task. Given the repetitiveness of the process and the fact that less than 1% of mammograms in the average screening population contain cancer, it's no wonder that a significant number of breast cancers--about 28%--are missed by radiologists. The fact that human error is such a significant obstacle makes mammography screening an ideal application for computer-aided detection (CAD) systems. CAD systems serve as a "second pair of eyes" to ensure that radiologists don't miss a suspect area on an image. They analyze patterns on a digitized mammographic image, identify regions that may contain an abnormality indicating cancer, and mark these regions. The marks are then inspected and classified by a radiologist. But CAD systems provide no diagnosis of any kind--it's up to the radiologist to analyze the marked area and decide if it shows cancer. In this Evaluation, we describe the challenges posed by screening mammography, the operating principles and overall efficacy of CAD systems, and the characteristics to consider when purchasing a system. We also compare the performance of two commercially available systems, iCAD's MammoReader and R2's ImageChecker. Because the two systems offer comparable sensitivity, our judgments are based on other performance characteristics, including their ease of use, the number of false marks they produce, the degree to which they can integrate with hospital information systems, and their processing speed. PMID- 12760159 TI - Inappropriate feeding caused by missing valve pins on Ross Flexiflo Quantum enteral feeding pumps. PMID- 12760160 TI - Imaging tables of International Surgical Systems C-arms can fall off. PMID- 12760161 TI - Advances in ischemic stroke epidemiology. PMID- 12760162 TI - Genetic and molecular epidemiologic methods for stroke. PMID- 12760163 TI - Genetic epidemiology of ischemic stroke. PMID- 12760164 TI - The role of inflammation in atherosclerosis. PMID- 12760165 TI - Magnetic resonance angiography. AB - MRA is a robust technique that still is undergoing modifications and improvements. As a noninvasive, or minimally invasive, technique using intravenous contrast injection, it offers complementary assessment of intracranial and extracranial arteries at the time of MRI, using similar coils and sequences. Images are similar to, but of lower resolution than, those of conventional angiography. PMID- 12760166 TI - Invasive angiography for arterial lesions of stroke. PMID- 12760167 TI - Intracranial atherosclerosis. PMID- 12760168 TI - Collateral circulation imaging for arterial lesions of stroke. PMID- 12760169 TI - Update on imaging aortic atherosclerosis. PMID- 12760170 TI - Advances in cerebrovascular ultrasound. PMID- 12760171 TI - Amyloidosis of cerebral arteries. PMID- 12760172 TI - Moyamoya disease. PMID- 12760173 TI - Cervical arterial dissection. PMID- 12760174 TI - Fibromuscular dysplasia. PMID- 12760175 TI - Vertebrobasilar disease. PMID- 12760176 TI - Small-vessel disease with lacunes. PMID- 12760177 TI - CADASIL. Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy. PMID- 12760178 TI - Biochemistry of ischemic stroke. PMID- 12760179 TI - Fifty years at Framingham: contributions to stroke epidemiology. AB - The author is obviously indebted to the many Framingham investigators and researchers over 35 years who have contributed to this remarkable enterprise. The reader is referred to the Framingham Study website (http://www.nhlbi.nih.gov/about/framingham/index.hm) for a more complete listing of publications and other background materials. PMID- 12760180 TI - Nutritional and metabolic aspects of stroke prevention. AB - Epidemiologic evidence, animal studies, angiographic and ultrasound studies in humans, and a limited number of clinical trials suggest that vitamins C and E may be protective and that folate, B6, and B12, by lowering homocysteine levels, may reduce stroke. However, these hypotheses require testing before widespread use of supplementary vitamins can be generally recommended (62). Clinical trials under way will test those hypotheses. In the meantime, it should be understood that the role of diet is much more important than is widely recognized. A diet low in saturated fat and cholesterol, low in sodium, high in potassium and calcium, and containing a lot of fruits and vegetables reduces blood pressure as much as an antihypertensive drug and in coronary patients is twice as effective as statin drugs in reducing death and myocardial infarction. Such a diet can therefore be confidently recommended as a source not only of natural proportions of vitamins and antioxidants but also for benefits that we are only beginning to define. PMID- 12760181 TI - Risk factor management to prevent stroke. PMID- 12760182 TI - An overview of nonseptic cardioembolic stroke. PMID- 12760183 TI - Specifics of patent foramen ovale. PMID- 12760185 TI - Cardiac rhythm disorders and muscle changes with cerebral lesions. PMID- 12760184 TI - Nonvalvular atrial fibrillation: an important cause of stroke. PMID- 12760186 TI - Sudden death. PMID- 12760187 TI - Cerebral venous and sinus thrombosis: incidence and causes. PMID- 12760188 TI - The treatment of cerebral venous sinus thrombosis. PMID- 12760189 TI - Reflections on the conduct of multicenter stroke prevention studies. PMID- 12760190 TI - Antithrombotic therapies for stroke prevention in atrial fibrillation. PMID- 12760191 TI - Anticoagulants for acute ischemic stroke. PMID- 12760192 TI - Anticoagulants for prevention of ischemic stroke: current concepts. PMID- 12760193 TI - Antithrombotic therapy for atherosclerotic intracranial arterial stenosis. PMID- 12760194 TI - Antithrombotic therapy in small subcortical strokes (lacunar infarcts). PMID- 12760195 TI - North American perspective of antiplatelet agents. PMID- 12760196 TI - Antiplatelet agents. European perspective. PMID- 12760197 TI - Treatment of symptomatic arteriosclerotic carotid artery disease. PMID- 12760198 TI - Treatment of asymptomatic arteriosclerotic carotid artery disease. PMID- 12760199 TI - Extracranial to intracranial bypass. PMID- 12760200 TI - Angioplasty and stenting. PMID- 12760201 TI - Choice and timing of specific stroke treatment. PMID- 12760202 TI - Intensive care management of specific stroke treatment. PMID- 12760203 TI - Hemicraniectomy for treatment of middle cerebral artery infarction. PMID- 12760204 TI - Intraarterial thrombolysis for acute ischemic stroke. PMID- 12760205 TI - Investigation by perfusion CT and diffusion-weighted MR imaging. PMID- 12760206 TI - Ongoing trials and future directions for acute ischemic stroke treatment. PMID- 12760207 TI - Medical complications during stroke rehabilitation. PMID- 12760209 TI - Partial body weight supported treadmill training for gait recovery following stroke. AB - PBWSTT has emerged from an interesting idea to a well-accepted and scientifically supported treatment modality for gait training after stroke. PBWSTT offers a task specific approach to functional gait retraining that is based on rehearsal of repetitive controlled gait cycles. Newer devices are being developed that will provide better mechanical control of the patient and reduce therapist effort. PMID- 12760208 TI - Cognitive rehabilitation of memory following stroke. Theory, practice, and outcome. AB - Impaired memory is a common consequence of ischemic stroke. Cognitive rehabilitation of memory is an essential component of any comprehensive rehabilitation program for these patients. Generalizable methods and methods to teach domain-specific knowledge are two principal means whereby impaired memory can be improved. Compensatory strategies can be taught to patients whose memory is not likely to improve. Cognitive remediation using these methods has been shown to be an effective intervention to treat memory deficits. More well controlled, randomized studies are needed to further investigate the efficacy of the various interventions available. PMID- 12760210 TI - Robot-aided sensorimotor training in stroke rehabilitation. AB - The first groups to use robotic devices to enhance the sensorimotor experience of patients recovering from upper limb paralysis after stroke have generated encouraging information. Patient acceptance and staff enthusiasm for robot training at Burke--MIT is consistently high. Robot training data are consistent with other controlled studies showing that more activity leads to more motor improvement and decreased impairment. Therapists equipped with a robotic device can increase the amount and intensity of movement of the paralyzed limb without sacrificing time spent training complex functionally appropriate kinematics. PMID- 12760211 TI - Poststroke depression. PMID- 12760213 TI - [SARS, globalization and Internet]. PMID- 12760212 TI - Pharmacotherapy in stroke rehabilitation. PMID- 12760214 TI - [A kidney donor should be treated like a king]. PMID- 12760215 TI - [Tragedies in drunk cells]. PMID- 12760216 TI - [Alternatives to sleeping pills for the elderly]. PMID- 12760217 TI - [Immunotherapy against psoriasis]. PMID- 12760218 TI - [Predictors for 21-year incidence of myocardial infarction in the Oslo study 1972/73]. PMID- 12760219 TI - [Pharmacological modulation of the inflammatory response after gunshot injuries in the pig]. PMID- 12760220 TI - [Chelating tests and dental amalgam fillings]. PMID- 12760222 TI - [Is current practice concerning living donors in kidney transplantations acceptable?]. PMID- 12760221 TI - [How are living kidney donors doing?]. PMID- 12760223 TI - [Deaths in police custody]. PMID- 12760224 TI - [Use of hypnotics in geriatric institutions in Hordaland and Sogn and Fjordane]. PMID- 12760225 TI - [Treatment of severe psoriasis with anti-TNF-alpha-antibody and methotrexate]. PMID- 12760226 TI - [Head-HUNT--hunting for causal headache factors]. PMID- 12760227 TI - [Wasp allergy vaccination]. PMID- 12760228 TI - [Neurological presentation of Churg-Strauss syndrome]. PMID- 12760229 TI - [Fatal carbon monoxide poisoning]. PMID- 12760230 TI - [Magnesium in pre-eclampsia/eclampsia]. PMID- 12760231 TI - [Disaster statistics with reservation]. PMID- 12760232 TI - [Organ donation in cooperation with relatives of the deceased]. PMID- 12760233 TI - [Wrong scientific techniques in health policy analyses]. PMID- 12760234 TI - [Cognitive therapy for palmar sweating and facial blushing]. PMID- 12760235 TI - [Untrustworthy criticism of an advertising brochure]. PMID- 12760236 TI - [A bitter pill--for whom?]. PMID- 12760238 TI - [Recruitment crisis in medical research]. PMID- 12760237 TI - [How should Norwegian drug cost policy be structured?]. PMID- 12760239 TI - [Why to keep the pediatric surgery specialty?]. PMID- 12760240 TI - Male oncology nurses share commitment, dedication and love of the profession. PMID- 12760241 TI - A comprehensive worksite cancer prevention intervention: behavior change results from a randomized controlled trial (United States). AB - OBJECTIVE: Workplace cancer prevention initiatives have been least successful with blue-collar workers. This study assesses whether an intervention integrating health promotion with occupational health and safety results in significant and meaningful increases in smoking cessation and consumption of fruits and vegetables, compared to a standard health promotion intervention, for workers overall and for blue-collar workers in particular. METHODS: A randomized controlled design was used, with 15 manufacturing worksites assigned to a health promotion (HP) or a health promotion plus occupational health and safety intervention (HP/OHS), and compared from baseline (1997) to final (1999). The response rates to the survey were 80% at baseline (n = 9019) and 65% at final (n = 7327). Both groups targeted smoking and diet; the HP/OHS condition additionally incorporated reduction of occupational exposures. RESULTS: Smoking quit rates among blue-collar workers in the HP/OHS condition more than doubled relative to those in the HP condition (OR = 2.13, p = 0.04), and were comparable to quit rates of white-collar workers. No statistically significant differences between groups were found for mean changes in fruits and vegetables. CONCLUSIONS: Integration of occupational health and safety and health promotion may be an essential means of enhancing the effectiveness of worksite tobacco control initiatives with blue-collar workers. PMID- 12760242 TI - Politics of prevention: expanding prevention benefits in the Medicare program. AB - It is critical that public health advocates understand the structure of the Medicare program and the impact of the political process on Medicare's benefit set. This article provides an overview of the design of Medicare and its explicit exclusion of prevention benefits in the original legislation. We then provide a history of subsequent legislation authorizing coverage of specific prevention benefits over the last twenty years. We critique the current process in light of innovation in preventive services and the influence of politics in the decision making. We conclude with a discussion of policy options to improve access to an appropriate range of evidence-based preventive services in Medicare within the context of new technology innovation and rising health care costs. PMID- 12760243 TI - Barriers to physical activity in the Hispanic community. AB - The purpose of this paper is to analyze barriers to leisure time physical activity in the Hispanic population in the U.S. Many studies have been conducted regarding this topic, primarily based on the general U.S. population. Fewer studies have concentrated specifically on Hispanics, where low levels of leisure time physical activity are reported. Research has usually stressed individual factors as barriers to exercise. A review of the literature addressing the different obstacles to physical activity will be presented within the context of immigration, poverty, and place of residency. This paper will attempt to explain why Hispanics tend to be less physically active and to make public health recommendations in order to minimize identified barriers. Suggestions and recommendations for future research that may help the understanding of the structural barriers will be provided. PMID- 12760244 TI - [Variation and evolution of meiosis]. AB - Meiosis arose in the evolution of primitive unicellular organisms as a part of sexual process. One type of meiosis, the so-called classical type, predominates in all kingdoms of eukaryotes. Meiosis is controlled by hundreds of genes, both shared with mitosis and specifically meiotic ones. In a wide range of taxa, which in some cases include kingdoms, meiotic genes and features obey Vavilov's law of homologous variation series. Synaptonemal complexes (SCs) temporarily binding homologous chromosomes at prophase I, ensure precise and equal crossing over and interference. SC proteins have 60-80% homology within the class of mammals but differ from the corresponding proteins in fungi and plants. Thus, nonhomologous SC proteins perform similar functions in different taxa. Some recombination enzymes in fungi and insects have common epitopes. The molecular mechanism of recombination is inherited by eukaryotes from prokaryotes and operates in special compartments: SC recombination nodules. Chiasmata, i.e., physical crossovers of nonsister chromatids, are preserved in bivalents until metaphase I due to local cohesion of sister chromatids in the remaining SC fragments. Owing to chiasmata, homologous chromosomes participate in meiosis I in pairs rather than individually, which, along with unipolarity of kinetochores (only in meiosis 1), ensures segregation of homologous chromosomes. The appearance of SC and chiasmata played a key role in the evolution of unicellular organisms since it promoted the development of a progressive type of meiosis. Some lower eukaryotes retain primitive meiosis types. These primitive modes of meiosis also occur in the sex of some insects that is heterozygous for sex chromosomes. I suggest an explanation for these cases. Mutations at meiotic genes impair meiosis; however, due to the preservation of archaic meiotic genes in the genotype, bypass metabolic pathways arise, which provide partial rescue of the traits damaged by mutations. Individual blocks of genetic program of meiotic regulation have probably evolved independently. PMID- 12760245 TI - [Genetic variation of nuclear ribosomal DNA of the insect order of cockroaches (Blattaria): phylogenetic analysis of restriction fragment length polymorphism]. AB - Species-specific characteristics of nuclear ribosomal DNA (rDNA) have been determined for the first time for six insect species of order Blattaria (Insecta, Dictyoptera) with the use of restriction analysis and Southern blotting. Probes containing highly conserved fragments of 18 S- and 28 S-like genes of the ribosomal RNA (rRNA) of Tetrahymena pyriformis were tested in this study. The genetic similarity tree constructed for the species studied agrees with evidence from classical taxonomy based on morphological, ecological, anatomical, and physiological characters. PMID- 12760246 TI - [Mutants of cyanobacterium Synechocystis sp. PCC6803 with insertion of the sodB gene encoding Fe-superoxide dismutase]. AB - The sodB gene encoding the only superoxide dismutase (Fe-SOD) in cells of the cyanobacterium Synechocystis sp. PCC6803 was inactivated with gentamycin resistance aacC1 marker insertions located in the direct or inverted order toward the sodB gene. The corresponding delta sodB12 and delta sodB22 mutants are characterized by the complete absence of superoxide dismutase activity and the loss of viability upon standard photoautotrophic cultivation. Mutant cells can grow under conditions of a decreased illumination intensity and upon addition of NaHCO3 with catalase or bovine serum albumin in the growth medium. The delta sodB22 mutant is auxotrophic for leucine due to the polar effect of insertion into the sodB gene on the downstream leuB gene controlling leucine biosynthesis. These data suggest that Fe-SOD is very important for providing tolerance of Synechocystis cells to oxidative stress and that sodB and leuB genes are organized into a single operon. PMID- 12760247 TI - [Osmotic shock induces expression of Vibrio fischeri lux genes in Escherichia coli cells]. AB - The effect of osmotic shock on the expression of genes in the lux regulon of marine bacteria Vibrio fischeri was studied in cells of Escherichia coli. Bioluminescence of cells was shown to drastically increase, when cells were exposed to osmotic shock at the early logarithmic growth phase. The expression of lux genes induced by osmotic shock is determined by the two-component regulatory system RcsC-RcsB. A nucleotide sequence in the regulatory region of the luxR gene homologous to the RcsB-box consensus of E. coli is assumed to be a primary site for this system. PMID- 12760248 TI - [Suppression of nonsense and frameshift mutations obtained by different methods for inactivating the translation termination factor eRF3 in yeast Saccharomyces cerevisiae]. AB - Mutations in genes of omnipotent nonsense suppressors SUP35 and SUP45 in yeast Saccharomyces cerevisiae encoding translation termination factors eRF3 and eRF1, respectively, and prionization of the eRF3 protein may lead to the suppression of some frameshift mutations (CPC mutations). Partial inactivation of the translation termination factor eRF3 was studied in strains with unstable genetically modified prions and also in transgenic yeast S. cerevisiae strains with the substitution of the indigenous SUP35 gene for its homolog from Pichia methanolica or for a recombinant S. cerevisiae SUP35 gene. It was shown that this partial inactivation leads not only to nonsense suppression, but also to suppression of the frameshift lys2-90 mutation. Possible reasons for the correlation between nonsense suppression and suppression of the CPC lys2-90 mutation and mechanisms responsible for the suppression of CPC mutations during inactivation of translation termination factors are discussed. PMID- 12760249 TI - [Pleiotropic effect of the hooded mutation of the rat on male fertility]. AB - Several reproductive parameters were studied in males homozygous (hh) or heterozygous (Hh) for the hooded mutation as compared with completely pigmented wild-type males (HH). Histological analysis of the testes was carried out in males of the three genotypes. The proportion of sterile males in homogeneous matings of homozygotes hh was twice as high as in matings of heterozygotes. The proportion of sterile males in matings yielding no progeny was also twice higher in homozygotes hh as compared with heterozygotes. No sterile males were detected in matings of completely pigmented wild-type animals. Unilateral cryptorchidism, a hypoplastic testis combined with a hyperplastic one, or hypoplasia of both testes were observed in some males homozygous for the hooded mutation. Morphologically, these defects were associated with underdevelopment or the complete absence of spermatogenic epithelium or with the presence of gaps and cells with large nondivided nuclei in the epithelium. The results showed that the hooded coat-color mutation exerts a pleiotropic effect on male fertility in rat. PMID- 12760250 TI - [Creation and genetic study of a collection of symbiotic mutants of the pea (Pisum sativum L.)]. AB - A collection of symbiotic pea mutants consisting of 34 lines was created. The morphobiological and symbiotic description of different types of pea mutants is presented. The mode of inheritance of symbiotic characters is analyzed. For the first time in pea, hypernodulation mutations with dominant inheritance were isolated. Differences in the structure of bacteroid tissues of root nodules were revealed between mutants with effective and ineffective nodules. Using reciprocal root/stem and stem/root grafts, the plant organ producing nodulation-controlling factors was established. New hypernodulation-controlling genes--nod4 and nod6- were identified. For the first time in pea, using pure lines derived from cultivars with contrasting nodulation, the new gene Nod5--nod5 was found. The dominant allele of this gene controls abundant nodulation, high nitrogen fixation, and nitrate resistance. Pea cultivars carrying the Nod5 gene that can be used in selection for improved symbiotic properties were identified. PMID- 12760251 TI - [Genome comparisons of three closely related flax species and their hybrids with chromosomal and molecular markers]. AB - Chromosome C-banding patterns were analyzed in three closely related flax species (Linum usitatissimum L., 2n = 30; L. angustifolium Huds., 2n = 30; and L. bienne Mill., 2n = 30) and their hybrids. In each case, the karyotype included metacentrics, submetacentrics, and one or two satellite chromosomes. Chromosomes of the three flax species were similar in morphology, size (1-3 microns), and C banding pattern and slightly differed in size of heterochromatic regions. In all accessions, a large major site of ribosomal genes was revealed by hybridization in the pericentric region of a large metacentric. A minor 45S rDNA site was observed on a small chromosome in L. usitatissimum and L. bienne and on a medium sized chromosome in L. angustifolium. Upon silver staining, a nucleolus organizing region (NOR) was detected on a large chromosome in all species. In L. angustifolium, an Ag-NOR band was sometimes seen on a medium-sized chromosome. In the karyotypes of interspecific hybrids, silver-stained rDNA loci were observed on satellite chromosomes of both parental species. RAPD analysis with 22 primers revealed a high similarity of the three species. The greatest difference was observed between L. angustifolium and the other two species. The RAPD patterns of L. bienne and L. usitatissimum differed in fewer fragments. A dendrogram of genetic similarity was constructed for the three flax species on the basis of their RAPD patterns. Genome analysis with chromosome and molecular markers showed that L. bienne must be considered as a subspecies of L. usitatissimum rather than a separate species. The three species were assumed to originate from a common ancestor, L. angustifolium being closest to it. PMID- 12760252 TI - [Macromutations and evolution: fixation of Goldschmidt's macromutations as species and genus characters. Papillomatosis and appearance of macrovilli in the rodent stomach]. AB - The cardiac portion of the stomach is lined with macrovilli in a few rodent genera. These are Mystromys (Cricetinae, Cricetidae), Myospalax (Myospalacinae, Cricetidae), Tachyoryctes (Tachyoryctinae, Cricetidae), and Cryptomys (Bathyergidae). The macrovilli favor the development of symbiotic flora and are called symbiovilli. Growth of the corneal epithelium of the cardiac portion of the stomach serves as a morphological basis of symbiovilli. Cases of a hereditary malignant neoplasm giving rise to the formation of multiple macrovilli in the cardiac portion of the stomach have been found in Microtus abbreviatus (Microtinae, Cricetidae), a vole endemic to St. Matthew Island in the Bering Sea. The macrovilli resulting from the papillomatosis mutation are morphologically and histologically identical to the macrovilli of the stomach of the four aforementioned genera. The voles affected with papillomatosis still survive long enough to reproduce. Therefore, the macromutation that leads to death in adult and old voles has been fixed as a species character in some rodent genera. At the early stages of papillomatosis, the pathogenic morphogenesis creates favorable conditions for the development of symbiotic microflora, which gives a selective advantage to the affected animals. It is assumed that mutations with pathogenic effects have been fixed as a species character as a result of heterochrony. The pathogenic neoplasm serves as a preadaptation for the growth of symbiotic flora in the stomach. The mechanisms of the fixation of Goldschmidt's "systemic mutations" during phylogeny are discussed. PMID- 12760253 TI - [Analysis of the polymorphic alleles of genes encoding phase 1 and phase 2 detoxication enzymes in patients with endometriosis]. AB - Polymorphysms of the three genes encoding phase 1 (CYP1A1, mEPH1, and CYP2E2) and the three genes encoding phase 2 (NAT2, GSTM1, and GSTT1) xenobiotic detoxication enzymes were typed by use of PCR in 74 patients with extragenital endometriosis. Distribution of the CYP1A1, mEPHX1, CYP2E1, NAT2, and GSTM1 polymorphic alleles in the patient group corresponded to that in the control group. At the same time, functionally defective genotypes GSTM1 0/0, NAT2 S/S; GSTM1 0/0, GSTT1 0/0; and GSTT1 0/0, NAT2 S/S were three, four and eight times more frequent among the patients than in healthy individuals. This observation suggests the existence of a distinct association between the functionally defective alleles of the phase 2 xenobiotic detoxication and endometriosis. Possible mechanisms underlying this association are discussed. It is suggested that typing of the NAT2, GSTM1, and GSTT1 genes can be useful for the assessment of the predisposition to endometriosis. PMID- 12760254 TI - [Gene frequencies and heterozygosity of the AB0 and RH blood group alleles in the populations of two cities of the Donetsk region, Ukraine]. AB - The frequencies of the AB0 and RH blood group alleles and heterozygosity indices were determined for the populations of two large industrial cities of Gorlovka and Mariupol. In the population of Gorlovka the gene frequencies were as follows: AB0*0 = 0.576, AB0*A = 0.266, AB0*B = 0.158, and RH*D = 0.592, in Mariupol the frequencies were AB0*0 = 0.584, AB0*A = 0.265, AB0*B = 0.151, and RH*D = 0.604. In Gorlovka the heterozygosity indices in respect to the AB0 and RH alleles were 0.572 and 0.483, respectively; in Mariupol, 0.566 and 0.478, respectively. There were no statistically significant differences between the two populations in respect to the genetic markers analyzed. However, the heterozygosity values obtained were more similar to the corresponding estimates for some populations of Russia, than for the total population of the Ukraine. PMID- 12760255 TI - [Gene insertion and deletion polymorphism in the serotonin transporter gene and personality traits measured by MMPI]. AB - Polymorphisms of the serotonin transporter gene are known to be associated with some personality traits measured by means of various psychological inventories. In the present work we attempted to find an association between genetic variants of serotonin transporter (loci VNTR-17 and 5-HTTLPR) and psychological traits scored by the MMPI inventory in 125 mentally healthy donors. No statistically significant differences in personality traits were found between carriers of different VNTR-17 genotypes. At locus 5-HTTLPR, significant between-genotype differences were revealed on the Schizophrenia scale (F = 3.49; P = 0.034) and on the validity scale F (F = 3.24; P = 0.042). The ss genotype carriers had the lowest scores on these scales. The score on the Psychopathic Deviate scale was significantly lower in the carriers of the ss genotype than in the combined group of the carriers of genotypes ll and ls (t = 2.07; P = 0.041). The differences on the validity scale K between the carriers of the ll and ss genotypes were also statistically significant (t = 2.49; P = 0.015). These results suggest that polymorphism of the serotonin transporter gene may be associated with the expression of schizoid traits (namely, social introversion, internal tension, weird thoughts and actions) in mentally healthy individuals. In the context of social adaptation, the personality profile configuration and data of statistical analysis indicate that the carriers of the ss genotype are more inclined to observe social norms than the carriers of the ll and ls genotypes. PMID- 12760256 TI - [Negative selection and computer models of the joint evolution of the patterns of polygenes, transposable elements, and origin identity labels]. AB - Computer simulation of the population dynamics of the genomic patterns of polygenes, transposable elements (TEs), and origin identity labels (OILs) in the course of negative selection for an additive quantitative trait has been performed. It was demonstrated that active polygene alleles disappear very rapidly, whereas the patterns of TEs and OILs continue their evolution determined by strict selective inbreeding and gene drift. Dendrograms of the patterns of polygenes, TEs, and OILs were constructed for all generations. It was demonstrated that the final consensus pattern of OILs consists of the fragments of the original patterns, which contain neither active polygene alleles nor modifier or marker TEs. Neutral TE copies were present in the final pattern, as should be expected in the case of gene drift. Inbreeding coefficient increased steadily but by generation 100 reached values higher than 0.9. All other parameters and initial conditions being the same, the responses to negative and positive selections were asymmetric. PMID- 12760257 TI - [Stabilizing selection and computer models of the joint evolution of patterns of polygenes, transposable elements, and origin identity labels]. AB - A computer model of the populations dynamics of the patterns of polygenes, transposable elements (TEs), and origin identity labels (OILs) in the course of stabilizing selection for an additive quantitative trait (with the target value being 0.4 of the maximum) was analyzed. It was demonstrated that the final plateaus of the trait value and the frequencies of the active values of polygenes are reached rapidly, namely, within five to seven generations (the effective selection period). The inbreeding coefficient during this period also grows rapidly and then gradually increases eventually reaching approximately 0.7. The inbreeding coefficient reaches plateau (at approximately 1.0) only in generations 300-350, which suggests the effect of gene drift. Dendrograms of the patterns of polygenes, TEs, and OILs were constructed for all generations. By generation 100 of selection, the final patterns of TEs and OILs were not formed completely. Fixations and losses, especially those of the OIL pattern, were delayed. In general, however, the population heterogeneity with respect to the patterns studied does not stabilize. This heterogeneity decreases the case of stabilizing selection, although more slowly than in the cases of positive and negative selections. PMID- 12760258 TI - [Temporal dynamics of ethnic assortativeness in the Tsivil'sk region of Chuvashia]. AB - Based on marriage records for 1950-1959, 1970-1979, and 1990-1999, ethnic assortativeness of Chuvash and Russian inhabitants of Tsivil'sk raion of the Chuvash Republic was calculated. It was shown that in half a century ethnic assortativeness of Chuvashes did not change (1.18, 1.15, and 1.15), while in Russians a clear downward trend was observed (4.20, 2.18, and 1.28). The data on ethnic assortativeness obtained were compared with those for the same populations from other regions of the Chuvashia examined earlier. PMID- 12760259 TI - [Inheritance of awn absence in tetraploid wheat species]. AB - Awn absence was shown to be inherited as a dominant character in the tetraploid wheat species Triticum dicoccum (Schrank) Schuebl. and T. durum Desf. but as a recessive one in T. aethiopicum Jakubz. The monogenic control of the character was demonstrated for all studied species. In accessions of emmer and durum wheat, the character is controlled by the dominant gene B1, located on chromosome 5A, and in Ethiopian wheat, by a recessive gene, which we designated as awn. The recessive awn gene was localized on chromosome 3B of T. aethiopicum with the use of D-genome disomic substitution lines of cultivar Langdon. PMID- 12760260 TI - [Comparative effects of rye chromosomes 1R and 5R on androgenesis in cultured anthers of wheat-rye substitution lines dependending on the line origin]. AB - The effects of rye chromosomes 1R and 5R on androgenesis in cultured anthers of wheat--rye substitution lines was studied as dependent on the cultivar origin of the rye chromosomes and on the wheat genome (A or D) subjected to substitution. Chromosome 1R stimulated embryogenesis in anther cultures, while chromosome 5R suppressed it regardless of whether the corresponding wheat chromosomes were substituted in the A or D genome. The effect of chromosome 1R on embryogenesis proved to depend on its cultivar origin. Along with rye chromosome 1R, wheat chromosome 1A was shown to substantially affect total seedling regeneration. Regeneration of green seedlings was dramatically affected both by rye chromosome 1R and by wheat chromosome 1D. The results supported the published data that individual androgenesis parameters (embryogenesis, total plant regeneration, green plant regeneration) are controlled by different genetic mechanisms. PMID- 12760261 TI - Airway obstruction after acute ozone exposure in BALB/c mice using barometric plethysmography. AB - BACKGROUND: Airway responsiveness after acute inhalation of ozone is related to the concentration and duration of ozone exposure. Using barometric whole-body plethysmography and increase in enhanced pause (Penh) as an index of airway obstruction, we measured the response of BALB/c mice to acute ozone inhalation to study the time course change of pulmonary function after ozone exposure. METHODS: Penh was measured before and after exposure to filtered air or 0.12, 0.5, 1, or 2 ppm ozone for 3 hr (n = 6/group). In addition, Penh was measured 24, 48 and 72 hr after ozone exposure. Bronchoalveolar lavage (BAL) and histopathologic examinations were performed. RESULTS: The increase in Penh after ozone exposure was significantly higher in the 0.12, 0.5, 1 and 2 ppm groups compared with the control group (all p < 0.01). Increases in Penh 24 hr after ozone exposure were significantly lower than those immediately after acute ozone exposure; however, increases in Penh 72 hr after ozone exposure were significantly higher than those in the control group (each p < 0.01). The proportion of neutrophils in BAL fluid was significantly higher in the group exposed to 2 ppm ozone than in the groups exposed to filtered air or 0.12 ppm ozone (both p < 0.01). CONCLUSION: These results indicate that airway obstruction is induced following ozone exposure in a concentration-dependent manner and persists for at least 72 hr. PMID- 12760262 TI - Changes in serum cytokine concentration: a morphological study of liver cirrhosis induced by common bile duct ligation in rats. AB - BACKGROUND: Liver cirrhosis is a diffuse hepatic fibrosis, and nodule formation. The transforming growth factor-beta 1 (TGF-beta 1) and interleukin-10 (IL-10) are very important cytokines in hepatic fibrogenesis. The aim of this study was to examine the relationship between the changes of the serum cytokines and morphological changes following common bile duct ligation in experimental rats. METHODS: Common bile ducts of fifty male Sprague-Dawley rats were ligated and seven male rats were set aside as controls. Five rats each were sacrificed in 1, 2, 4, 6, 8, and 10 experimental weeks. Light microscopic studies and liver function tests were performed during the above experimental weeks. The levels of serum TGF-beta 1 and IL-10 were analyzed by ELISA. Also, alpha smooth muscle actin (alpha-SMA) immunohistochemical stains were performed. RESULTS: On the eighth week after common bile duct ligation, most hepatic lobular areas had been replaced by proliferated bile ducts and fibrous tissue (typical biliary cirrhosis). Serum TGF-beta 1 levels between the control group and the common bile duct ligation group showed statistically significant changes. The alpha-SMA was stained at proliferated bile ducts. These findings were correlated with each other. CONCLUSION: Thus, this experiment may clarify our understanding of the mechanism in liver fibrogenesis. Also, indicated is a need to explore the therapeutic potential of these cytokines as anti-fibrotic agents. PMID- 12760263 TI - Prognostic value of AML 1/ETO fusion transcripts in patients with acute myelogenous leukemia. AB - BACKGROUND: The t (8;21) (q22;q22), which produces the fusion gene AML1/ETO, is associated with relatively good prognosis and, in particular, with a good response to cytosine arabinoside. Analysis of t (8;21) positive leukemic blasts has shown characteristic morphological and immunological features. We performed this study to investigate the incidence of AML1/ETO rearrangement in adult acute myelogenous leukemia (AML), especially in M2 subtype, to make a comparison of clinical, morphological and immunophenotypic characteristics between AML1/ETO rearrangement positive and negative group in patients with AML and to analyze the correlation with other biological parameters. METHODS: From May 1995 to Sept. 2000, fifty-nine patients with AML, including twenty-nine AML-M2, were studied. RNAs were extracted from leukemic cells and reverse transcriptase mediated polymerase chain reaction (RT-PCR) for AML1/ETO fusion transcript was done. Chromosome study, immunophenotypic and clinical characteristics were analyzed and statistical analysis was done. RESULTS: The incidence of AML1/ETO fusion transcripts was 22.0% in AML and 44.8% in AML-M2. The morphologic finding of bone marrow in AML-M2 showed higher incidence of Auer rods, large blast with prominent golgi and abnormal granules in AML1/ETO positive patients. There was no significant difference of immunophenotype. AML patients with AML1/ETO had a tendency of higher complete remission rate (81.8% vs 56.6%, p = 0.13). The overall survival (median; 82.2 weeks vs 34.4 weeks, p = 0.02) and progression free survival (median; 50.9 weeks vs 20.4 weeks, p = 0.02) of AML1/ETO positive group were longer than those of the negative group in AML. AML-M2 patients with AML1/ETO rearrangement had also a tendency of longer overall survival and progression free survival, although there was no significant difference between both groups. CONCLUSION: Our data suggest that AML1/ETO rearrangement is detected frequently in AML, especially M2, and is a favorable prognostic factor. Thus, molecular diagnostic approaches should be used routinely to identify patients with this gentic subtype of AML. PMID- 12760264 TI - Treatment outcome of adult acute lymphocytic leukemia with VPD(L) regimen: analysis of prognostic factors. AB - BACKGROUND: Because of the relative paucity of data regarding the clinical outcome in adult patients with acute lymphocytic leukemia (ALL) in Korea, we analyzed clinical courses in adult ALL patients treated with VPD (L) regimen (vincristine, prednisolone, daunorubicin, L-asparaginase) at the Seoul National University Hospital, and evaluated prognostic factors influencing the outcome. METHODS: Patients with ALL newly diagnosed between October 1994 and June 2000 at our hospital were analyzed retrospectively. Fifty-three patients were evaluable. Induction chemotherapy consisted of VPD with (46 cases) or without L-asparaginase (7 cases). After complete remission (CR), consolidation therapy, CNS prophylaxis and maintenance chemotherapy were administered. RESULTS: Ages ranged from 16 to 67 (median 30). CR rate was 86.8% (46/53) and no significant prognostic factor was found for the CR rate. With a median follow-up time of 27.2 months (range 12.9-83.0 months) in living patients, the median overall survival (OS) for all cases was 16.7 months (13.4-20.1 months, 95% C.I.) and the estimated 4-year OS rate was 25.4% +/- 8.9%. The median relapse-free survival (RFS) was 12.2 months (8.4-16.0 months, 95% C.I.), and 3-year RFS rate was 29.9% +/- 10.2%. Poor prognostic factors for OS were Ph chromosome (p = 0.005) and T-cell immunophenotype (p = 0.03). For RFS they were Ph chromosome (p = 0.01) and the presence of a mediastinal mass (p = 0.03). CONCLUSION: Despite an initial excellent response to the VPD (L) regimen, newer therapeutic strategies, including more intensive postremission therapies, are urgently needed because of the high relapse rate. Future therapeutic approaches need to be stratified according to several prognostic factors. PMID- 12760265 TI - The role of C-reactive protein on the long-term clinical outcome after primary or rescue percutaneous coronary intervention. AB - BACKGROUND: We examined the clinical and angiographic outcomes, success rate of the percutaneous coronary intervention (PCI) and long-term survival rate after primary or rescue PCI in patients with acute myocardial infarction (AMI) according to the level of the C-Reactive Protein (CRP) on admission. METHODS: Two hundred and eight consecutive patients with AMI who underwent primary or rescue PCI between 1997 and 1999 at Chonnam National University Hospital were divided into two groups: Group I (n = 86, 59.9 +/- 9.3 years, male 74.4%) with a normal CRP (< 1.0 mg/dL, mean value = 0.43 +/- 0.14 mg/dL) on admission and Group II (n = 122, 59.1 +/- 10.4 years, male 83.6%) with an elevated CRP (> or = 1.0 mg/dL, mean value = 3.50 +/- 0.93 mg/dL) on admission. RESULTS: There were no significant differences in teh baseline characteristics noted between the two groups. The incidence of cardiogenic shock was higher in Group II than in Group I (Group I; 3/86, 3.5% vs. Group II; 15/122, 12.3%, p = 0.026). The coronary angiographic findings did not differ between the two groups. The ejection fraction and Thrombolysis In Myocardial Infarction flow grade improved after PCI in both groups. The primary success rate of PCI was 94.2% (81/86) in Group I and 95.1% (116/122) in Group II (p = 0.776). The survival rates for Group I were 97.7%, 97.7% and 96.5%, and those for Group II were 91.8%, 91.0% and 86.9% at 1, 6 and 12 months, respectively (p = 0.043 at 1 month, p = 0.040 at 6 months, p = 0.018 at 12 months). CONCLUSION: A high incidence of cardiogenic shock and worse long-term survival after PCI are observed in AMI patients with an elevated CRP. PMID- 12760267 TI - Primary non-Hodgkin's lymphoma of the bladder with bone marrow involvement. AB - Involvement of the lower urinary tract by advanced non-Hodgkin's lymphoma (NHL) has been reported in up to 13% of cases, but primary NHL of the urinary bladder is very rare. A 35-year-old man was admitted to our hospital with a chief complaint of gross hematuria with left flank pain on April 12, 2001. Cystoscopy revealed an edematous broad-based mass on the left lateral wall of the bladder, and transurethral biopsy showed NHL, diffuse large B-cell type. Abdomino-pelvic CT scan demonstrated left-side hydronephrosis and hydroureter with left proximal ureter infiltration and thickening of the left lateral wall of the bladder with perivesical fat infiltration without lymph node enlargement. Full-scale staging work-up revealed the bone marrow as the solely involved site. The lesions of the bladder and left urinary tract were nearly completely regressed after two cycles of systemic cyclophosphamide, doxorubicin, vincristine and prednisone (CHOP) chemotherapy with simultaneous restoration of urinary function. PMID- 12760266 TI - The role of cardiopulmonary exercise test in mitral and aortic regurgitation: it can predict post-operative results. AB - BACKGROUND: We evaluated the efficacy of the cardiopulmonary exercise test as an objective indicator of functional status and as a pre-operative prognostic indicator in patients with mitral regurgitation (MR) and aortic regurgitation (AR). METHODS: Cardiopulmonary exercise tests and echocardiography were performed in 47 patients (MR: 30, AR: 15, MR + AR: 2) before surgery and repeated one year after surgery. We compared the New York Heart Association (NYHA) functional class, peak oxygen consumption rate (VO2peak), exercise duration, left ventricular dimension and ejection fraction, before and after surgery. RESULTS: Initial VO2peak and exercise duration were significantly different according to NYHA class. A year later, NYHA functional class improved from 2.1 +/- 0.1 to 1.4 +/- 0.1 (p < 0.001). The VO2peak was significantly increased (21.7 +/- 1.0 to 23.7 +/- 1.0 mL/kg per min, p = 0.008) and exercise duration also increased (521.7 +/- 35.9 to 623.3 +/- 35.7 seconds, p < 0.001). When patients were analysed according to their post-operative NYHA functional class, those with class I showed significantly different pre-operative VO2peak (class I: 23.7 +/- 1.1, II: 18.3 +/- 1.5 mL/kg per min, p = 0.005) and exercise durations (class I: 587.5 +/- 43.2, II: 415.6 +/- 55.7 seconds, p = 0.02). Patients with higher pre operative VO2peak (19.0 mL/kg per min) more frequently became NYHA functional class I than those with a lower pre-operative VO2peak (76.7% vs. 35.3%, p = 0.02). But baseline left ventricular dimension and ejection fraction by echocardiography were not different between post-operative class I and II group. CONCLUSION: VO2peak and exercise duration are excellent parameters to evaluate the subjective functional class and to predict the post-operative functional class of patients with MR and/or AR. Patients with a pre-operative VO2peak of 19.0 mL/kg per min or more will have a better functional status one year after surgery. PMID- 12760269 TI - Hypersensitivity to mosquito bites associated with natural killer cell-derived large granular lymphocyte lymphocytosis: a case report in Korea. AB - Hypersensitivity to mosquito bites (HMB) is characterized by intense skin reactions at bite sites. The pathogenesis of HMB might be related to clonal lymphoproliferation of Epstein-Barr virus DNA-positive natural killer (NK) cells. We report the first case of HMB possibly associated with NK cell-derived large granular lymphocyte (NK-LGL) lymphocytosis in Korea. PMID- 12760268 TI - A large malignant insulinoma: case report with endosonographic, immunohistochemical and ultrastructural features. AB - Malignant insulinoma in the beta cells of the pancreatic islet is rare and usually presented as hypoglycemia. We report a case of large malignant insulinoma in a 53-year-old Korean woman. A presumptive clinical diagnosis was made before surgery, based on the high plasma insulin-to-glucose ratio and a large solitary heterogeneous pancreatic mass by abdominal computed tomography and endosonography. The tumor measured 5.8 x 4.7 x 4.5 cm in dimension and showed capsular invasions and metastases in two of four peripancreatic lymph nodes. The tumor cells were strongly immunoreactive to insulin and had a high Ki-67 labeling index (13%) and atypical membranous electron-dense granules, ranging from 120 to 400 nm in diameter, in the cytoplasm on electron microscopy. The patient was treated by distal pancreatectomy with splenectomy and rapidly recovered without neurohypoglycemic symptoms. This case showed not only lymph node metastases, the most reliable parameter for malignancy in pancreatic endocrine tumors, but also other valid diagnostic clues, such as high Ki-67 labeling index, heterogeneous enodosonographic findings, capsular invasions with large tumor and pure atypical secretory granules. PMID- 12760270 TI - A case of simultaneous xanthogranulomatous cholecystitis and carcinoma of the gallbladder. AB - Xanthogranulomatous cholecystitis (XGC) is a rare inflammatory disease of the gallbladder. Not only does XGC occasionally present as a mass formation with adjacent organ invasion like a malignant neoplasm, it can also infrequently be associated with gallbladder cancer. In the situation, it is difficult to make a differential diagnosis between the diseases. Here, we describe a case of a simultaneous XGC and a carcinoma of the gallbladder in a 61-year-old woman. To the best of our knowledge, there are only a small number of reports on this combination of diseases. PMID- 12760275 TI - Kentucky Nurses' Association and Kentucky Nurses Foundation--vision for the future of nursing. PMID- 12760271 TI - Hydronephrosis by an aberrant renal artery: a case report. AB - Ureteropelvic junction obstruction is usually intrinsic and is most common in children. Aberrant renal arteries are present in about 30% of individuals. Aberrant renal arteries to the inferior pole cross anteriorly to the ureter and may cause hydronephrosis. To the best of our knowledge, although there are some papers about aberrant renal arteries producing ureteropelvic junction obstruction, there is no report of a case which is diagnosed by the new modalities, such as computed tomography angiogram (CTA) or magnetic resonance angiogram (MRA). We describe a 36-year-old woman with right hydronephrosis. Kidney ultrasonogram and excretory urogram revealed right hydronephrosis. CTA and MRA clearly displayed an aberrant renal artery and hydronephrosis. The patient underwent surgical exploration. For the evaluation of hydronephrosis by an aberrant renal artery, use of CTA and MRA is advocated. PMID- 12760272 TI - Very severe aplastic anemia appearing after thymectomy. AB - Aplastic anemia is a rare complication of thymoma and is extremely infrequent after thymectomy. We present a case of a 60-year-old woman with very severe aplastic anemia appearing sixteen months after thymectomy for a thymoma. She underwent thymectomy for a thymoma in April 2000. Preoperative examination revealed no hematologic abnormality. About sixteen months after the operation, she was readmitted because of pancytopenia with cough and fever. Bone marrow aspiration revealed a very severe hypoplasia in all the three cell lines with over 80% fatty tissue, and chest CT revealed no recurrence of thymoma. Her aplastic anemia had responded to cyclosporine A and granulocyte-colony stimulating factor (G-CSF). PMID- 12760276 TI - Have you missed an essential part of your retirement plan? Planning for the risk of long-term care. PMID- 12760280 TI - To stain or disdain, that is the question.... PMID- 12760281 TI - Contribution of IgH-PCR to the evaluation of B-cell lymphoma involvement in paraffin-embedded bone marrow biopsy specimens. AB - We investigated whether the determination of clonality by polymerase chain reaction (PCR) analysis of immunoglobulin heavy chain (IgH) gene rearrangements could be helpful in the evaluation of B-cell lymphoma (BCL) involvement of bone marrow (BM) biopsy specimens. We evaluated 83 paraffin-embedded BM biopsy specimens from 26 patients with BCL. When BM biopsy specimens considered positive, "suspicious," or negative by morphologic and immunohistochemical examination were evaluated by PCR, a monoclonal B-cell population was detected in 81% (39/48), 64% (9/14), and 11% (2/18), respectively. In most cases, a reproducible monoclonal IgH gene rearrangement was observed from BM and extramedullary sites. Nevertheless, in 4 cases, a different and independent monoclonal IgH rearrangement was observed during the disease course. PCR is efficient and complementary to morphologic and immunohistochemical examination for the evaluation of BCL involvement of BM biopsy specimens, especially when a reproducible rearrangement is found in 2 different samples. PMID- 12760282 TI - Lineage-specific identification of nonhematopoietic neoplasms by flow cytometry. AB - To extend flow cytometry (FC) to the diagnosis of nonhematopoietic neoplasms, we have developed new flow cytometric assays to identify expression of cytokeratin, epithelial cell adhesion molecule (EpCAM)/epithelial glycoprotein-2, myogenin, and CD99. To validate these assays, we correlated the flow cytometric results with the histologic and immunohistochemical results on paraffin-embedded tissue in a series of 21 cases, including 17 carcinomas, 1 atypical carcinoid, 2 rhabdomyosarcomas, and 1 Ewing sarcoma/primitive neuroectodermal tumor (ES/PNET). Six of 7 assayed carcinomas and the carcinoid were positive for cytoplasmic cytokeratin by the flow cytometric assay. EpCAM was expressed by 11 of 12 carcinomas that were assayed by FC. Both rhabdomyosarcomas expressed myogenin by FC, and the ES/PNET case expressed CD99. Interestingly, the blast-associated antigen CD90 was expressed uniformly on the ES/PNET case and on subsets of cells in the rhabdomyosarcoma and carcinoma cases. Potential applications of the flow cytometric assay to nonhematopoietic neoplasms will include evaluating samples with limited material, monitoring disease persistence and recurrence in patients with previous diagnoses, and making rapid diagnoses in urgent cases. PMID- 12760283 TI - Elimination of instrument-driven reflex manual differential leukocyte counts. Optimization of manual blood smear review criteria in a high-volume automated hematology laboratory. AB - Automated peripheral blood leukocyte differential counts (LDCs) are widely accepted in routine practice. However, many laboratories still reflexively perform manual LDCs based solely on abnormal automated results or instrument "flags," before any manual triage step. We describe our transition to a procedure that uses manual methods to validate, rather than to replace, automated LDCs (an approach recommended early in the development of automated methods, but still not used in many clinical laboratories). Manual microscopic scans were performed in lieu of manual LDCs. Each scan that revealed cell types not quantifiable by the instrument triggered a manual LDC. However, if the manual scan simply confirmed the cell types seen on automated LDC, then the automated result was released, even if clinically significant quantitative abnormalities were present. This policy reduced manual LDCs by more than 70% and was validated by a manual retrospective audit. Patient care and laboratory operations can be optimized by using manual microscopic examination as a validation procedure rather than as a reflexive substitute for automated methods. There is no clinical rationale for reflex performance of manual LDCs based solely on instrument warnings. PMID- 12760285 TI - Prognostic impact of acute myeloid leukemia classification. Importance of detection of recurring cytogenetic abnormalities and multilineage dysplasia on survival. AB - To evaluate the prognostic impact of acute myeloid leukemia (AML) classifications, specimens from 300 patients with 20% or more bone marrow myeloblast cells were studied. Specimens were classified according to the French American-British Cooperative Group (FAB), the World Health Organization (WHO), the Realistic Pathologic Classification, and a cytogenetic risk group scheme. Cases with fewer than 30% blast cells did not have a 5-year survival significantly different from cases with 30% or more blast cells, and survival was similar for the low blast cell count group and cases with multilineage dysplasia and 30% or more blasts. Categories of AML with recurrent cytogenetic abnormalities of t(15;17), t(8;21), inv(16)/t(16;16), and 11q23 showed significant differences in 5-year survival. No significant difference was identified between AMLs arising from myelodysplasia and de novo AMLs with multilineage dysplasia, but all cases with multilineage dysplasia had a worse survival than all other AMLs and other AMLs without favorable cytogenetics. FAB types M0, M3, and M4Eo showed differences in survival compared with all other FAB types, with M0 showing a significant association with high-risk cytogenetics and 11q23 abnormalities. Other FAB groups and WHO AML, not otherwise categorized subgroups did not show survival differences. These findings suggest that the detection of recurring cytogenetic abnormalities and multilineage dysplasia are the most significant features of current AML classification. PMID- 12760284 TI - Immunohistochemical detection of VEGF in the bone marrow of patients with acute myeloid leukemia. Correlation between VEGF expression and the FAB category. AB - We studied vascular endothelial growth factor (VEGF) expression in bone marrow sections obtained from 3 healthy donors and 41 patients with acute myeloid leukemia (AML) of various French-American-British (FAB) subtypes by immunohistochemical analysis using an anti-VEGF antibody. In normal bone marrow, the anti-VEGF antibody reacted with myeloid progenitor cells and megakaryocytes but not with erythroid cells or mature granulocytic cells. High levels of VEGF were found in the bone marrow in patients with AML-M1, -M2, -M3, -M4, -M4Eo, and M5. In these leukemias, the vast majority of myeloblasts (> 90%) expressed VEGF. By contrast, in AML-M0, the percentage of VEGF-positive blasts was lower in most cases (median, 42%), and if at all detectable, these blast cells contained only trace amounts of VEGF. In AML-M3 and -M4Eo, maturing granulocytes failed to express VEGF similar to granulocytes in normal bone marrow. In AML-M6, myeloblasts exhibited VEGF, whereas erythroid cells did not. In AML-M7, blast cells and megakaryocytes were identified as major sources of VEGF. In summary, VEGF expression in the bone marrow is restricted to certain stages of differentiation and maturation of myeloid cells and correlates with the FAB category. PMID- 12760286 TI - Thermodynamic determination of plasma and leukocyte beta-hexosaminidase isoenzymes in homozygote and heterozygote carriers for the GM2 gangliosidosis B1 variant. AB - In the GM2 gangliosidosis B1 variant, the mutated isoenzyme A of beta hexosaminidase (Hex) is incapable of hydrolyzing ganglioside GM2 and negatively charged substrates. Biochemical characterization of this lysosomal disease is carried out using synthetic alpha-subunit-specific sulfated substrates, as heat inactivation assays are not applicable. The apparent enzyme activation energy of Hex using the chromogenic substrate 3,3'-dichlorophenolsulfonphthaleinyl N-acetyl beta-D-glucosaminide is related directly to the relative proportions of Hex A and Hex B isoenzymes. This thermodynamic variable was used for the study of Hex enzyme heterogeneity in 3 patients with the GM2 gangliosidosis B1 variant and 6 heterozygote carriers. Hex activity was determined at 25 degrees C, 30 degrees C, 35 degrees C, and 37 degrees C in a Cobas Bio analyzer (Roche Diagnostics, Basel, Switzerland), and Arrhenius plot slopes and apparent activation energies were calculated in plasma samples and mononuclear and polymorphonuclear leukocyte lysates. The determination of the Hex isoenzymes in plasma presented a high discrimination power for B1 variant patients but not for heterozygote carriers, in whom false-negative results may be obtained. However, thermodynamic evaluation of the isoenzyme composition of Hex in leukocyte lysates permits the biochemical identification of patients with the GM2 gangliosidosis B1 variant and of heterozygote carriers. PMID- 12760287 TI - Performance characteristics of four immunonephelometric assays for the quantitative determination of IgA and IgM in cerebrospinal fluid. AB - Measurement of IgA and IgM in cerebrospinal fluid (CSF) can be useful in the diagnosis of multiple sclerosis and other central nervous system disorders. The Dade Behring (Deerfield, IL) N Latex IgA and N Latex IgM tests on the BN II System and Beckman Coulter (Brea, CA) low-concentration IgA and IgM tests on the IMMAGE Immunochemistry System were evaluated for linearity, imprecision, method comparison, and reference interval verification. Both IgA methods were linear from 1.4 to at least 50 mg/L. Both IgM methods were linear from 0.14 to more than 6 mg/L. The total imprecision of the BN II IgA and IgM methods and the IMMAGE IgA method was less than 10%. The imprecision of the IMMAGE IgM method was 10.2% at 0.49 mg/L and less than 5% at higher IgM concentrations. Method comparison studies indicated that IgA and IgM methods on both instruments showed good comparability. Reference interval studies demonstrated that both methods had similar reference intervals that agreed with published values of less than 6 mg/L for IgA and less than 1.3 mg/L for IgM. Methods for quantifying IgA and IgM in CSF on the BN II and IMMAGE nephelometers perform well and give comparable results. PMID- 12760288 TI - p63 and TTF-1 immunostaining. A useful marker panel for distinguishing small cell carcinoma of lung from poorly differentiated squamous cell carcinoma of lung. AB - We studied the usefulness of p63 and thyroid transcription factor-1 (TTF-1) immunostains for differentiating poorly differentiated squamous cell carcinoma (PDSCC) from small cell lung carcinoma (SCLC). We used monoclonal antibodies reactive to p63 or TTF-1 to stain 4-microns-thick sections from 30 formalin fixed, paraffin-embedded lung biopsy and resection specimens and 7 alcohol-fixed, formalin-postfixed, paraffin-embedded cell blocks from lung fine-needle aspirations (FNAs). For p63, we used a streptavidin-biotin kit, diaminobenzidine as the chromogen, and a hematoxylin counterstain. We used automated immunostaining for TTF-1. The 37 cases included 23 SCLCs, 13 PDSCCs, and 1 carcinoma initially diagnosed as PDSCC. All 23 SCLCs were negative or, rarely, equivocal for p63; 20 (87%) of 23 were TTF-1+; nuclear staining ranged from strong and/or frequent to weak and/or uncommon. All 13 PDSCCs were TTF-1-/p63+ with intense staining of 50% to 100% of tumor cells. One case originally diagnosed as PDSCC was TTF-1+/p63-, suggestive of SCLC; after morphologic reexamination and immunostaining for neuroendocrine markers, it was reclassified as intermediate-type SCLC. TTF-1 immunostaining showed equal or increased sensitivity in alcohol-fixed cytologic cell block samples compared with formalin fixed biopsy material; in 1 SCLC case, the biopsy specimen was TTF-1-; however, the FNA cell block stained positively. p63 and TTF-1 appear to be useful for differentiating SCLC from lung PDSCC in formalin-fixed and alcohol-fixed, formalin-postfixed material. PMID- 12760289 TI - Endoscopic ultrasound-guided fine-needle aspiration findings of gastrointestinal leiomyomas and gastrointestinal stromal tumors. AB - True leiomyomas of the gastrointestinal system are rare but remain the most common mesenchymal tumors of the esophagus. It has become important to distinguish these tumors from gastrointestinal stromal tumors (GISTs) because the neoplasms have different prognoses and treatment options. We describe and compare clinical findings and the following fine-needle aspiration (FNA) features of 9 gastrointestinal leiomyomas and 19 GISTs sampled with endoscopic ultrasound: overall cellularity, cell group features, cell shape and cytoplasmic features, nuclear characteristics, background, cell block features, and immunohistochemical results. Gastrointestinal leiomyomas and GISTs have different clinical and cytologic features that help pathologists distinguish these tumors, and the immunohistochemical findings that help define these lesions can be derived readily from cell block material obtained by endoscopic ultrasound-guided FNA. PMID- 12760290 TI - Terminal diffuse alveolar damage in relation to interstitial pneumonias. An autopsy study. AB - Acute exacerbations of idiopathic pulmonary fibrosis/cryptogenic fibrosing alveolitis (IPF/CFA) are rare and typically terminal events, but their relationship to underlying patterns of idiopathic interstitial pneumonias is unknown. We reviewed autopsy material from patients who died of diffuse alveolar damage in the clinical setting of pulmonary fibrosis, both idiopathic and with background fibrosing alveolitis with connective tissue disorders (FA-CTDs), and compared them with cases of acute interstitial pneumonia. Of 15 patients with acute exacerbations of IPF/CFA (n = 12) or FA-CTD (n = 3), 12 had a background pattern of usual interstitial pneumonia and 3 had fibrotic nonspecific interstitial pneumonia. All cases of fibrotic nonspecific interstitial pneumonia were seen in association with FA-CTD. The cause of acute exacerbations is unknown, but our data suggest that toxic effects of oxygen and triggering infection are unlikely causes. In patients with CTDs, it remains uncertain whether the acute exacerbation is related to the fibrosis, the associated CTD, or a combination of these factors. Acute exacerbations of IPF/CFA may be a more common terminal event than previously thought. PMID- 12760291 TI - MIB-1 and DNA topoisomerase II alpha could be helpful for predicting long-term survival of patients with glioblastoma. AB - Approximately 10% of patients with glioblastoma survive more than 2 years after diagnosis. Distinguishing these patients from those who died within 2 years of diagnosis is clinically significant. We studied the MIB-1 labeling index (LI) and DNA topoisomerase II alpha LI of glioblastomas from 34 patients who lived for more than 2 years after diagnosis and of glioblastomas from 34 age- and sex matched control patients who died within 2 years of diagnosis. The means of MIB-1 and topoisomerase II alpha LIs of the group with a better outcome were lower. With 35 as the cutoff point for the MIB-1 LI and 26 as the cutoff point for the topoisomerase II alpha LI, both MIB-1 and topoisomerase II alpha LIs were related significantly to survival. Our study showed that both MIB-1 and topoisomerase II alpha could help predict long-term survival of patients with glioblastomas. Multivariate analyses revealed that MIB-1 was a better prognostic marker than topoisomerase II alpha. PMID- 12760292 TI - Apoptosis regulation differs between ulcerative colitis-associated and sporadic colonic tumors. Association with survivin and bcl-2. AB - To clarify kinetics in ulcerative colitis (UC)-associated lesions, cell proliferation, apoptosis, and expression of apoptosis-inhibitory proteins were studied. Ki-67 labeling and survivin and bcl-2 expression were examined immunohistochemically in 22 low-grade dysplasias (LGDs), 25 high-grade dysplasias (HGDs), and 13 adenocarcinomas associated with UC, and for comparison in 21 sporadic adenomas with LGD, 22 sporadic adenomas with HGD, and 21 invasive adenocarcinomas. Apoptosis was studied with nick-end labeling and immunohistochemical analysis of single-stranded DNA. In UC-associated LGDs, Ki-67 -positive cells were more frequent in the lower than the upper half of the crypt, related to bcl-2 expression, while in sporadic adenomas such cells were more common in the upper half. No difference in apoptosis was found between UC associated LGDs and sporadic adenomas with LGD or between UC-associated HGDs and sporadic adenomas with HGD. However, UC-associated carcinomas exhibited a lower apoptotic count than their sporadic invasive counterparts. This seemed related to higher survivin expression without a significant difference between the 2 types of invasive lesions regarding bcl-2 levels. Apoptosis is less frequent in UC associated than in sporadic invasive colon carcinomas, this being linked to elevated survivin expression. The control of apoptosis may be different in the 2 types of tumorigenesis. PMID- 12760293 TI - Granulomatous eccrinotropic lymphomatoid papulosis. AB - We describe 9 patients with a novel variant of lymphomatoid papulosis characterized by prominent localization of the infiltrate around the eccrine coil, resulting in nodular expansion of the coil accompanied by variable granulomatous inflammation. Light microscopy, immunohistochemical analysis using antibodies to CD2, CD3, CD4, CD5, CD7, CD8, and CD30 in 6 cases, and polymerase chain reaction--single-stranded conformational polymorphism analysis for T-cell receptor gamma chain gene rearrangement in 5 cases revealed 2 cytomorphologic patterns (large cell dominant with polymorphous inflammation, small cell lymphocyte--rich with an inconspicuous large cell component [phenotypes, CD30+/CD3+/CD4+/CD7-for large atypical cells; reactive for small lymphocytes]) and clonal restriction in 4 and polyclonality in 1 of the lymphocyte-dominant cases. During an average 6-year follow-up, no lymphomas developed. Recognition of this variant is important--accentuation of the infiltrate around the eccrine coil and cutaneous nerves, presence of granulomatous inflammation, dominance of small lymphocytes in the dermis, and variable extension into the panniculus may lead to diagnostic confusion with entities such as discoid lupus erythematosus, Jessner lymphocytic infiltrate of skin, subcutaneous T-cell lymphoma, and persistent arthropod bite reaction. Our findings suggest that pruritus, a younger age at diagnosis, and a more indolent course are defining clinical features. PMID- 12760294 TI - Cause of death and sudden cardiac death after heart transplantation. An autopsy study. AB - Postmortem findings in 39 patients following cardiac transplantation are presented. Causes of death were right-sided heart failure after transplantation (6 [15%]), infection (5 [13%]), multisystem organ failure (4[10%]), complications of noncardiac surgery (3 [8%]), acute rejection (3 [8%]), malignant neoplasm (3 [8%]), graft vascular disease (3 [8%]), preservation procurement injury (3 [8%]), cardiac arrhythmia (2 [5%]), other (4 [10%]), and unclear (3 [8%]). Seven patients in medically stable condition died after a sudden cardiac arrest, and these constituted 27% (7/26) of deaths more than 1 month after transplant. The 7 sudden cardiac arrests were due to graft vascular disease (2 [29%]), acute rejection (2 [29%]), cardiac arrhythmias (2 [29%]), and hyperkalemia during an exacerbation of acute renal failure (1 [14%]). In 3 of the 5 patients who died of sudden cardiac arrest not due to acute rejection, hemodialysis and plasmapheresis were triggers of the event. Pulmonary hypertensive arteriopathy was associated with early death and right-sided heart failure, and 6 of 8 patients with these changes died perioperatively or postoperatively. PMID- 12760295 TI - The neuropathology of West Nile virus meningoencephalitis. A report of two cases and review of the literature. AB - West Nile virus (WNV) is an emerging mosquito-transmitted encephalitis virus first recognized in North America in 1999. The pathologic manifestations of WNV infection have not been well defined. This study documents the clinicopathologic features, including autopsy findings, of 2 cases: an 81-year-old man who contracted WNV infection with meningoencephalitis and a polio-like paralysis and a hospitalized 74-year-old woman with meningoencephalitis who acquired WNV through transfusion. The pathologic findings in both cases were marked by perivascular and leptomeningeal chronic inflammation, microglial nodules, and neuronophagia, predominantly involving the temporal lobes and brainstem. These findings also were present in the spinal cord, especially the lumbar region, of the patient with polio-like paralysis. In both cases, most of the inflammatory infiltrate was composed of CD3+ T lymphocytes (a predominance of CD8+ over CD4+ T cells), CD68+ macrophages, and rare CD20+ B lymphocytes. These cases further define the clinical and pathologic spectrum of central nervous system disease in WNV infection. PMID- 12760299 TI - Anger management. PMID- 12760298 TI - Queensland nurses assist women in South Africa to respond to AIDS. PMID- 12760300 TI - Active service. PMID- 12760301 TI - The HSJ interview: Gill Morgan. Open season. Interview by Nick Edwards. PMID- 12760302 TI - Foundation trusts. Power sharing. AB - Foundation trusts will in theory have greater freedoms over pay, non-pay rewards and recruitment, although details are still unclear. Foundations are likely to differ in their use of these powers, partly because of desire and partly because of practical limits. Taking advantage of new powers for short-term gain, at the expense of other trusts, will have negative effects if not part of a wider development of HR practice. PMID- 12760303 TI - Local recruitment. Just the job. AB - Local deprived communities can be a rich recruitment source, but will be difficult to access using traditional means. Long-term unemployed people and school children often have poor perceptions of working in the NHS. It is important to combine recruitment with training opportunities for existing staff. Funding for recruitment programmes can be accessed from a variety of sources. PMID- 12760304 TI - Diversity. Chapter and diverse. AB - Neglecting diversity and equality issues in recruitment and people development can be dangerous, both in legal and human costs. Collating material into a training pack, which could be delivered by non-experts, helped spread best practice. Launching the material with a conference raised its profile and helped ensure that it would be used well. PMID- 12760305 TI - Education. Teach to his own. AB - The Teaching Company Scheme allows subsidized employment of a graduate on a fixed term contract. TCS is gradually being taken up by NHS organisations although it can be challenging to reconcile with its traditional focus on private sector criteria. South Tyneside's project involved developing a healthy living and working strategy. PMID- 12760306 TI - Work-life balance. The hours. PMID- 12760307 TI - Data briefing. Severe acute respiratory syndrome. PMID- 12760308 TI - Steroid and xenobiotic receptor (SXR) is a key system for the acquisition of cisplatin resistance in endometrial cancer cells. AB - A novel steroid and xenobiotic receptor (SXR) may be involved in chemoresistance, and we studied this receptor in endometrial cancer cell lines. The cisplatin (CDDP)-sensitive Ishikawa cell line and its CDDP-resistant sub-clone (ISIW+) were used. ISIW+ cells showed a much higher SXR expression. When Ishikawa cells were cultured with SXR anti-sense oligonucleotides (AS), the cells failed to pass crisis and did not gain cisplatin resistance. In an experiment using SCID mice, all AS-treated animals survived, whereas controls had 50% survival at 35 days. The present data indicate that SXR is a key system to induce, maintain and reverse a cisplatin-resistant phenotype in endometrial cancer cells. PMID- 12760309 TI - Expression of interleukin 6 and its receptor in human gastric and colorectal cancers. AB - Interleukin 6 (IL-6) is a pleiotropic cytokine with many physiological functions. The present study was designed to determine the expression of IL-6 and its receptor (IL-6R) in human gastric and colorectal cancers. Nine gastric- and nine colorectal cancer cell lines were analysed. The IL-6 gene was expressed in two gastric cancer cell lines and one colorectal cancer cell line; however, most of the cancer cell lines studied expressed the IL-6R gene. The level of IL-6 secretion in the gastric cancer cell lines correlated with the level of soluble IL-6R secretion, and was significantly higher (< approximately 100 pg/ml) than the level of IL-6 secretion in the colorectal cancer cell lines (< approximately 50 pg/ml). These results suggest that IL-6 may act in a paracrine fashion rather than an autocrine fashion in these cell lines. PMID- 12760310 TI - The diagnostic value of troponin T and myoglobin levels in acute myocardial infarction: a study in Turkish patients. AB - This study compares the diagnostic value of troponin T (TnT) and myoglobin with creatinine kinase (CK) for myocardial infarction (MI) in a tertiary care centre in a developing nation. The study group comprised 33 acute myocardial infarction patients and 27 healthy controls. Receiver operating characteristic curves for TnT, myoglobin and CK were drawn and areas under the curve calculated. At admission, myoglobin levels had greater diagnostic sensitivity than TnT or CK levels. After 2 h, myoglobin and TnT had equal sensitivity and specificity, whereas CK still had lower sensitivity than myoglobin and TnT. After 4 h there was no difference between the tests. It was concluded that myoglobin levels on admission and TnT at 2 h had the greatest diagnostic rate, whereas all the tests were similar after 4 h for MI. PMID- 12760311 TI - Does adenotonsillectomy improve growth in children with obstructive adenotonsillar hypertrophy? AB - Faltering growth may be associated with adenotonsillar hypertrophy, but its pathophysiological mechanism is unclear. This study included 29 pre-pubertal children with obstructive adenotonsillar hypertrophy, and aimed to investigate the probable difference in energy intake and serum insulin-like growth factor 1 (IGF-1) and insulin-like growth factor binding protein 3 (IGFBP-3) levels before and 6 months after adenotonsillectomy. Weight and height standard deviation scores, energy intake per kilogram and serum IGF-1 levels were found to be significantly higher 6 months after adenotonsillectomy, indicating that adenotonsillar hypertrophy is associated with poor growth. PMID- 12760312 TI - A single ascending dose study of epigallocatechin gallate in healthy volunteers. AB - This randomized, double-blind, placebo-controlled study assessed the safety, tolerability and plasma kinetic behaviour of single oral doses of 94% pure crystalline bulk epigallocatechin gallate (EGCG) under fasting conditions in 60 healthy male volunteers. In each group of 10 subjects, eight received oral EGCG in single doses of 50 mg, 100 mg, 200 mg, 400 mg, 800 mg or 1600 mg, and two received placebo. Blood samples were taken at intervals until 26 h later. The area under the concentration-time curve from 0 h to infinity (AUC(0-infinity)), the maximum plasma concentration (Cmax) of EGCG, the time taken to reach the maximum concentration (Tmax), and the terminal elimination half-life (t1/2z) of EGCG were determined. Safety and tolerability were assessed. In each dosage group, the kinetic profile revealed rapid absorption with a one-peak plasma concentration versus time course, followed by a multiphasic decrease consisting of a distribution phase and an elimination phase. The mean AUC(0-infinity) of total EGCG varied between 442 and 10,368 ng.h/ml. The according mean Cmax values ranged from 130 to 3392 ng/ml and were observed after 1.3-2.2 h. The mean t1/2z values were seen between 1.9 and 4.6 h. Single oral doses of EGCG up to 1600 mg were safe and very well tolerated. PMID- 12760314 TI - The effects of prednisolone and serum malondialdehyde levels in puppies with experimentally induced meconium aspiration syndrome. AB - The aim of this study was to investigate the effect of different doses of prednisolone in puppies experimentally induced with meconium aspiration syndrome (MAS). Meconium was collected from human babies in the first day of life and was released into the trachea of 11 newborn puppies to induce MAS. Puppies were treated with 2 mg/kg prednisolone (standard dose), 30 mg/kg prednisolone (megadose) or 0.9% saline, all administered intravenously. The study ended 20 h after meconium aspiration and the lungs were then scored for histopathology. Animals not treated with prednisolone deteriorated after 8 h while respiration rate, oxygenation, pH and partial pressure of carbon dioxide values were better in the prednisolone-treated groups. Histopathology scores were better in the treatment groups compared with the control group, with megadose giving the best result. At the end of the study, serum malondialdehyde levels were significantly higher in the megadose prednisolone group compared with the other two groups. In conclusion, we determined that prednisolone reduced physiological and histological changes in puppies with MAS and that a 30 mg/kg dose was more effective than 2 mg/kg. PMID- 12760313 TI - Celecoxib is as efficacious as naproxen in the management of acute shoulder pain. AB - This study compared the analgesic efficacy and safety of the cyclo-oxygenase-2 specific inhibitor celecoxib with the non-specific non-steroidal anti inflammatory drug, naproxen, in patients with acute shoulder pain. Adult patients with shoulder pain, onset within the previous 14 days and shoulder pain of > or = 40 mm on a 100 mm visual analogue scale (VAS), were treated with oral doses of celecoxib or naproxen for 14 days and followed by a visit at day 42 in a double blind, randomized study. The primary efficacy assessment was change in maximum pain at rest on a 100 mm VAS at day 14 compared with baseline. In addition, secondary efficacy pain and functional assessments were analysed at baseline, day 14 and day 42. A total of 202 patients were included in the trial (99 celecoxib 400 mg/day; 103 naproxen 1 g/day). The difference in change from baseline at day 14 in maximum pain at rest was not statistically significant between the two treatment groups, but was numerically higher for celecoxib than for naproxen ( 47.9 +/- 2.5 versus -42.3 +/- 2.5, respectively). According to the limits of the 95% confidence interval of the difference between groups (-12.52; 1.38), celecoxib appeared to be at least as effective as naproxen. All secondary efficacy measures followed the same pattern, showing similarity between the two treatments with a trend in favour of celecoxib. The incidences of adverse events were similar for both groups. Fewer patients experienced epigastric pain with celecoxib (seven patients versus 14 with naproxen). This adverse event led to discontinuation in two patients receiving celecoxib and five receiving naproxen. Celecoxib 400 mg/day was at least as effective as naproxen 1 g/day in managing pain in this condition. PMID- 12760315 TI - Relationship between mandibular nutrient canals and hypertension. AB - This study was undertaken to investigate the correlation between the presence of nutrient canals in the mandibular anterior region and hypertension. Periapical radiographs taken from the mandibular anterior region of 139 normotensive and 76 hypertensive edentulous patients were examined. There was no statistical difference in the incidence of nutrient canals between normotensive and hypertensive patients, suggesting that the presence of nutrient canals is not indicative of the existence of hypertension. PMID- 12760316 TI - Effect of galanin on plasma glucose, insulin and pancreatic glucagon in dogs. AB - The effect of synthetic galanin on plasma glucose, insulin and pancreatic glucagon levels in dogs was studied. Infusion of galanin caused a rapid, reversible and dose-dependent reduction in basal insulin level. A maximal increase in blood glucose level accompanying the insulin decrease was observed when galanin was administered at a dose of 4 micrograms/kg per h. Pancreatic glucagon levels showed little change compared with basal secretion. These results indicate that galanin is involved in the regulation of glucose through control of insulin secretion. PMID- 12760318 TI - Epidemiological features of ulcerative colitis in Trakya, Turkey. AB - To determine the epidemiological features of ulcerative colitis in the Trakya region of Turkey, southeast Europe, we conducted a descriptive, cross-sectional, hospital-based study. All subjects were followed, and age, sex, place of residence, family history, educational status, tobacco consumption and use of oral contraceptives were recorded. The study included 49 cases of ulcerative colitis. The incidence of the disease was 0.59 per 100,000 in 1998, 0.89 per 100,000 in 1999, 0.89 per 100,000 in 2000 and 0.69 per 100,000 in 2001. The overall prevalence of the disease was 4.9 per 100,000; it was 2.18 per 100,000 in rural areas and 5.87 per 100,000 in urban areas. As in the Mediterranean countries, both the incidence and the prevalence of ulcerative colitis were found to be low. The incidence was significantly higher in urban areas than in rural areas. PMID- 12760317 TI - Identification of three strains of Mycobacterium species isolated from clinical samples using fatty acid methyl ester profiling. AB - The cellular fatty acid profiles of 67 strains belonging to three different species of the genus Mycobacterium were determined by gas chromatography of the fatty acid methyl esters, using the MIDI Sherlock Microbial Identification System (MIS). The species M. tuberculosis, M. xenopi and M. avium complex were clearly distinguishable and could be identified based on the presence and concentrations of 12 fatty acids: 14:0, 15:0, 16:1 omega 7c, 16:1 omega 6c, 16:0, 17:0, 18:2 omega 6.9c, 18:1 omega 9c, 18:0, 10Me-18:0 tuberculostearic acid, alcohol and cyclopropane. Fatty acid analysis showed that there is great homogeneity within and heterogeneity between Mycobacterium species. Thus the MIS is an accurate, efficient and relatively rapid method for the identification of mycobacteria. PMID- 12760319 TI - A therapy-resistant chronic leg ulcer treated successfully with topical basic fibroblast growth factor. AB - We report a patient with rheumatoid arthritis who had a chronic leg ulcer with long-standing resistance to therapy. Topical basic fibroblast growth factor (bFGF) at a dose of 30 micrograms/day for 3 months was clinically effective in reducing the ulcer area by promoting angiogenesis, re-epithelization, granulation and scar formation. bFGF may be an effective treatment for therapy-resistant leg ulcers. PMID- 12760321 TI - Damages for "unwanted" healthy child awarded to physically disabled mother but not to mentally handicapped mother. PMID- 12760320 TI - Transient blindness following hysteroscopy. AB - The potential adverse effects resulting from absorption of irrigation fluids during endoscopic procedures are well documented. Glycine, which is commonly used as an irrigation solution, has an inhibitory effect both on the central nervous system and on the retinal cells. We report the case of a woman who developed transient blindness following hysteroscopic myomectomy in which glycine was used as the irrigation solution. PMID- 12760322 TI - Photography and vision: impressionism or reality? PMID- 12760323 TI - Reforms in the Criminal Law. PMID- 12760324 TI - The abbreviated multidimensional acculturation scale: empirical validation with two Latino/Latina samples. AB - Two studies were conducted to develop and examine internal consistencies and validate the Abbreviated Multidimensional Acculturation Scale. Study 1 participants were 156 Latino/Latina college students. Findings indicated good internal reliabilities for all 3 subscales. Adequate concurrent validity was established with length of residence in the United States. The scale also showed adequate convergent and discriminant validity. Study 2 participants were 90 Latino/Latina community members. The subscales were also reliable and showed adequate concurrent validity with length of residence in the United States. Convergent and discriminant validity were also adequate. Construct validity was further demonstrated through factorial analyses of the combined samples (N = 246). Three separate factors emerged: cultural identity, language competence, and cultural competence. PMID- 12760325 TI - Chinese immigrant mothers of adolescents: self-perceptions of acculturation effects on parenting. AB - Immigrant families in the United States experience many adjustments, including the challenge of parenting in a new context. The relationship between self perceptions of parenting behavior and mothers' perceived acculturation levels for themselves and their children was examined in 95 Chinese immigrant mothers. Data showed that mothers perceived their children as more acculturated than themselves, mothers reported a great need to learn more about how their children are growing up today, a larger perceived acculturation gap was associated with more parenting difficulties, and several factors contributed to perceptions of a more favorable parenting experience. Culturally responsive methodologies used in the translation and administration of measures, and specific modifications made to develop the Chinese version of the Parent Success Indicator, are discussed. PMID- 12760326 TI - Discrimination and Puerto Rican children's and adolescents' mental health. AB - Two studies of Puerto Rican youths' development on the U.S. mainland examined the consequences of perceived racial/ethnic discrimination on mental health. In Study I, children were found to have a low likelihood of perceiving discrimination, whereas in Study 2, nearly half of the adolescent sample reported perceiving racial/ethnic discrimination. Although both groups scored high on multiple indicators of mental health, perceiving discrimination and worrying about discrimination were negatively associated with some dimensions of self-esteem and positively associated with depression and stress. Adolescents were aware of negative stereotypes about Puerto Ricans, and nearly half of them related discriminatory instances. Results suggest that both perceiving discrimination and anxiety regarding discrimination can serve as risk factors for the mental health of this population. PMID- 12760327 TI - A qualitative study of adaptation experiences of 1.5-generation Asian Americans. AB - Adaptation experiences of 1.5-generation Asian American college students (N = 10) were examined using the consensual qualitative research method. Results indicated 4 domains of adaptation experiences: preimmigration experiences, acculturation and enculturation experiences, intercultural relationships, and support systems. Participants reported that English proficiency played a significant role in their initial adjustment. Currently, most of the participants reported feeling identified with both the U.S. and Asian cultures. Some participants reported having experienced racism in the past. Many participants noted that they currently have no difficulty establishing friendships with culturally different persons. Participants reported currently feeling most close to friends of a similar background and that they usually seek support from friends, family, and religious organizations, but not from a psychologist or counselor. PMID- 12760328 TI - Stress within a bicultural context for adolescents of Mexican descent. AB - Folkman and Lazarus's theory of stress and coping was used to develop a measure assessing the perceived stress within a bicultural context. Middle school students of Mexican descent (N = 881) reported their perceived stress from intergenerational acculturation gaps, within-group discrimination, out-group discrimination, and monolingual stress. Although immigrant youths reported more total number of stressors, U.S.-born youths reported more stress from needing better Spanish and impact of parents' culture. Immigrant youths reported more stress from needing better English in school. Higher stress was associated with more depressive symptoms for both U.S.-born and immigrant youths. Although this study has identified some elements of stress, it has not identified positive coping mechanisms of the bicultural context for Latino youths. PMID- 12760329 TI - Culture and validity of the Symptom Checklist-90-Revised and Profile of Mood States in a New Zealand student sample. AB - New Zealand students' performance was examined on assessments of psychopathology and mood as compared to normative data from the United States. New Zealand university students (N = 137) completed the Symptom Checklist-90-Revised (SCL-90 R) and Profile of Mood States (POMS). Mean performances differed significantly from normative data for each SCL-90-R scale. No significant differences were found for the POMS scales. Within the sample, European (n = 82), Maori (n = 24), and Asian (n = 24) participants differed significantly on SCL-90-R obsessive compulsive, phobic anxiety, and anxiety scales and POMS scales of tension and confusion. Implications for assessment of New Zealand samples are discussed. PMID- 12760330 TI - Feeling frugal: socioeconomic status, acculturation, and cultural health beliefs among women of Mexican descent. AB - Psychosocial and socioeconomic variables are often confounded. The authors combined quantitative with grounded theory analysis to investigate influences of acculturation, socioeconomic status (SES), and cultural health beliefs on Mexican descent women's preventive health behaviors. In 5 focus group interviews sampling across levels of acculturation and SES, women expressing more traditional Mexican health beliefs about breast cancer screening were of lower SES and were less U.S. acculturated. However, SES and acculturation were uncorrelated with screening behaviors. Qualitative analysis generated hypotheses about joint influences of SES and traditional health beliefs; for example, low-SES women may learn frugal habits as part of their cultural traditions that influence their health care decision making, magnifying SES-imposed structural restrictions on health care access. PMID- 12760331 TI - [In memory of Maria Orlando: physical and psychological health in menopause]. AB - Over the last fifty years life expectancy has increased by some 15 years for both sexes, causing an overall increase in the elderly. This will lead in the near future, to a number of economic, social and health problems, particularly in developing countries, especially to women whose life expectancy, in general, is longer than men's. Indeed, globally, already at the end of the twentieth century more that 60% of the population over 75 years of age were women. Among the elderly, and in particular among women in the third and fourth age, age-specific pathologies jeopardize the quality of life of many. In the long run, there are two types of problems that greatly affect women: cardiovascular and neoplastic diseases. Today, especially in industrialized countries, hormone replacement therapy is in widespread use, even though problems connected with the long-term use of estrogens are still to be fully understood. In particular recent controlled clinical trials have questioned the effectiveness of estrogens in preventing cardiovascular diseases. In conclusion, the years after menopause that in some cases can exceed 40 years, represent a new challenge for medicine and society; this is a challenge that must be met by giving precedence to human values, such as economic and social solidarity. PMID- 12760332 TI - [Reproductive health of immigrant women in the Lazio region of Italy]. AB - The number of legal immigrant women living in the Lazio region of Italy has increased in recent years; there were 69,320 "permessi di soggiorno" (temporary residence permits) issued in 1992 but 119,778 issued in 2000. This analysis concerns women who left hospital after having a live birth, an induced abortion or a spontaneous abortion. The foreign women were compared with all (Italian and foreign) women living in Lazio. The general fertility rate is slightly higher among all women resident in Lazio than among the foreign women, but the latter have a higher rate at ages 18-24 years. The risk of induced abortion is higher (34.8/1000 women) among the foreigners than among all residents (10.5/1000 women). The spontaneous abortion ratio was also higher among the foreigners (213.8/1000 live births) than the residents (154.6/1000 live births). There is clear evidence of an increase in the demand from foreign women for medical services for reasons associated with reproduction. The provision of these services needs to be modified to take account of their particular needs. PMID- 12760333 TI - [Access to public health services and HIV prevention: experimentation of participated planning with adolescents]. AB - The present study reports strategies and approaches found by adolescents through participated planning aiming at facilitating access of adolescents to HIV prevention services. Meetings and focus groups were carried out in secondary schools to collect information on attitudes, perceptions and opinions of adolescents on the way they use health services, the factors which are seen as obstacles to access, and the best strategies to overcome such obstacles. Analysis of the information gathered was carried out using content analysis. Among other results gathered it was evident that the social representation of AIDS was the major factor which underlay the decision of adolescents to avoid these specific health services and that taking part in participated planning allowed them to feel less distant to this illness which they initially felt did not concern them. PMID- 12760335 TI - Modulation of the multidrug resistance (MDR) phenotype in CEM MDR cells simultaneously exposed to anti HIV-1 protease inhibitors (PI's) and cytotoxic drugs. AB - Vinblastine, vincristine and doxorubicyn are currently used in chemotherapeutic treatments of several malignancies including HIV-1 associated tumours Kaposi's sarcoma (KS) and non-Hodgkin lymphoma (NHL). Hence, AIDS patients also affected by KS and NHL may be simultaneously subjected to highly active antiretroviral therapy (HAART) and cytotoxic drugs to combat HIV-1 infection and cancer aggressiveness. In order to assess if the combination of these therapies may affect cell growth and survival of P-glycoprotein expressing MDR variants of the human CD4+ T-lymphoblastoid CEM cell line, the protease inhibitors (PI's) ritonavir, saquinavir and indinavir were tested in an in vitro assay for their ability to potentiate the vinblastine, vincristine and doxorubicyn cytotoxicity. The results we obtained demonstrated that at the concentration of 10 micrograms/ml, ritonavir and in a lesser extent saquinavir act as MDR reversing agents. By contrast, the PI indinavir at least in the CEM cell system, does not affect the patterns of drug resistance. The level of chemosensitization exerted by ritonavir and saquinavir suggests that these PI's may render P-glycoprotein expressing MDR cells de novo susceptible to the antineoplastic drugs vinblastine, vincristine and doxorubicyn. PMID- 12760334 TI - [HIV prevention and adolescents: a sample survey on Italian high school students]. AB - This article resulted from a survey carried out by some researchers of the Telefono Verde AIDS (TVA-National AIDS Help-Line) of the Istituto Superiore di Sanita, with the collaboration of some colleagues of the Segreteria per le Attivita Culturali. It is also a summary of strategies of intervention against HIV/AIDS for young people implemented by the Italian Ministry of Health. The study was developed within the context of the workshops on HIV/AIDS education carried out since 1995 by some researchers of TVA. These workshops fall within some other initiatives arranged by the Segreteria per le Attivita Culturali for the diffusion of scientific and technological information. Our study aimed at assessing the level of knowledge on HIV/AIDS among some different high-school students. Data processing points out a widespread and quite good level of knowledge on HIV/AIDS in general and particularly on ways of transmission and prevention measures. Results obtained could not be generalized because the sample involved in the survey represented a privileged target, already sensitive to the issues concerning HIV infection and AIDS dealt with in the context of specific teaching programmes. PMID- 12760337 TI - [Use of phage libraries for the in vitro production of recombinant monoclonal antibodies of predetermined specificity]. AB - The biotechnological generation of monoclonal antibodies of predetermined specificity has traditionally involved the production of hybridomas obtained by somatic cellular fusion of splenocytes from immunized animals with myeloma cell lines bearing selectable markers. Now, monoclonal antibodies could be genetically engineered thus bypassing all the natural systems for making antibodies. Filamentous bacteriophages provides a means to display and select large single chain fragments variable (scFv) repertoires created by cloning the natural rearranged V-immunoglobulin genes or introducing predetermined level of randomization into germline V-gene segments. In this article we demonstrated that by using a well characterized scFv phage synthetic library it is possible to generate in vitro recombinant human monoclonal antibodies directed to a large array of antigens showing different molecular weights, conformations and origins. PMID- 12760336 TI - Advanced glycosylation end product quantification: differently produced polyclonal antisera do not share the recognition of epitopes of different nature. AB - Advanced glycosylation end products (AGE) which are probably involved in the pathogenesis of diabetic complications, comprise a series of related chemical structures. Thus different antisera might recognize particular AGE epitopes rather than the complete range of epitopes. To test this hypothesis, two antisera were raised using different immunization techniques and different AGE-carrier proteins as immunogens. The antisera reactivity towards different AGE-proteins under various experimental conditions was compared. Both antisera recognized all AGE-proteins, although with different binding curves. Following pre-incubation with carboxymethyllysine-BSA (CML-BSA) (an oxidation-derived AGE) one antiserum partially retained its reactivity, suggesting recognition of non-oxidation derived AGE. This result was confirmed both in the cross-reactivity and the preincubation experiments and when the reactivity of the antisera was tested against antigens incubated under oxidative and non-oxidative conditions. These results confirmed the hypothesis that differently produced antisera may not share the recognition of epitopes of different nature and suggest the necessity to adopt a standardized methodology for the production of antisera for an accurate and reproductible determination of the in vivo AGE concentration. PMID- 12760338 TI - Investigation into the role of apolipoprotein B gene 8344C/T variant on plasma cholesterol levels by allele-specific PCR amplification. AB - The 8344C/T polymorphism of the apoB gene was genotyped by an original modification of PCR allele-specific amplification consisting in a single amplification reaction double-primed by two opposite allele-specific oligonucleotides nested in a larger amplified fragment. This method was used to genotype 200 randomly selected healthy individuals (113 males, 87 females). The frequency of the rare allele in this random Italian population was 0.240, i.e. not far from the 0.282 frequency observed in hypocholesterolemic Norwegians and suggestive of a moderating effect on LDL levels of our population. However, we did not find any significant cholesterol-lowering effect of this polymorphism either by comparing the frequency of mutant alleles in the population stratified for its plasma lipoprotein levels or by studying the association between ApoB genotype and the different lipoproteins. In conclusion this ApoB polymorphism appeared to have a secondary role in LDL- and HDL-cholesterol variations of our population. PMID- 12760339 TI - Testing insecticide susceptibility of Phlebotomus perniciosus and P. papatasi (Diptera: Psychodidae) in Italy. AB - A study was undertaken to establish the susceptibility of two recently colonized sand fly species, Phlebotomus perniciosus and P. papatasi to some insecticides. A laboratory colony of P. papatasi, unexposed to insecticides for a period of ten years, was used as reference strain. Bioassay tests were carried out according to the WHO standard protocols based on the mortality rates recorded at different exposure times. Serial (from 1 to 60 min) exposures to DDT 2%, lambda-cyhalothrin 0.06% and permethrin 0.2% were tested on groups of 30 adult sugar-fed female sand flies, for a total of 7,470 specimens. Data were subjected to probit analysis and the susceptibility was calculated at LT50 and LT95 by comparison with the laboratory reference strain. The LT50 values for P. perniciosus were 19.9, 3.2 and 6.9 min, and for P. papatasi 18.0, 7.4 and 11.0 min, using DDT, lambda cyhalothrin and permethrin, respectively. The results showed that two Italian populations of P. perniciosus and P. papatasi from Campania region and from Rome, respectively, are susceptible to the insecticides tested as compared with the reference strain used. PMID- 12760340 TI - A neurophysiological approach to effects induced by accelerated particles on the central nervous system in mice. AB - This experiment was designed to study mechanisms of "phosphene vision". A beam of accelerated carbon particles (energy 100 and 300 MeV/n) was delivered into retina and sensorimotor cortex of anaesthetised mice bearing chronic electrodes. Cycles of light flashes were also administered. Following accelerated particles stimulation cycles to the retina, 500 to 5000 per stimulus, 30 stimuli per cycle, stimulus time-related brain waveforms in mice cortex were recorded. Their amplitude and latency appeared to be related to the energy delivered. The short waveforms latency (approximately 30 ms), and the lack of retinic potentials and oscillatory potentials following particles stimuli suggest that the impact of accelerated particles does not produce activation of retinic chemical structures. PMID- 12760341 TI - Erratum to "Sex differences in gamma-glutamyltransferase in people aged 40-42 years in two Norwegian counties". AB - Gamma-glutamyltransferase (GGT) is widely used as a marker of alcohol intake, although it is documented that other factors are also associated with serum levels of GGT. The total population of men and women aged 40-42 years in two Norwegian counties was invited to participate in a health survey program. GGT was measured in 8116 men and 8689 women--67 % of the eligible population. In sex specific multiple regression analyses, GGT showed a positive association with body mass index (BMI), cholesterol, ln triglycerides, systolic blood pressure, and number of drinks per 2 weeks for both men and women. Glucose and 'years of smoking' were significant in women only. Cups of boiled coffee per day and physical activity in spare time were inversely associated with GGT level for both men and women. A significant positive interaction between alcohol intake and BMI was observed for men but not for women. The use of GGT as a marker of alcohol consumption in middle-aged persons should take into account sex, BMI and drinking of boiled coffee. PMID- 12760342 TI - [DMW Walter Siegenthaler Prize 2002. Epidemiologic study on Creutzfeldt-Jakob disease wins the award]. PMID- 12760343 TI - [What should the anaesthetist know about current laws on medical products?]. PMID- 12760344 TI - [Progress of gene therapy in pancreatic carcinoma]. PMID- 12760345 TI - [Functional effects of phosphorylation and acetylation modification of p53]. PMID- 12760346 TI - [Advances in mechanism of chromosomal translocations in soft tissue sarcoma]. PMID- 12760347 TI - [Significance of minimal residual leukemia detection of acute leukemia during remission induction]. PMID- 12760348 TI - Demyelinating encephalitis. PMID- 12760349 TI - Early manifestations of testicular dysgenesis in children: pathological phenotypes, karyotype correlations and precursor stages of tumour development. AB - Testicular dysgenesis derives from abnormal gonadal development caused by chromosome aberrations/mosaicisms or mutations/deletions in SRY or other genes responsible for testicular differentiation. Dysgenetic male pseudohaermaphroditism has bilateral dysgenetic testes characterized by a cortical network of anastomosing seminiferous cords that penetrate a thin albuginea. In asymmetric gonadal differentiation (or Mixed Gonadal Dysgenesis) a dysgenetic testis associates with a streak gonad with primitive sex cords embedded in an ovarian-like stroma. Uni- or bilateral ovotestes identify true haermaphroditism. Fluorescent in situ hybridisation studies demonstrate that the sex chromosomes of mosaic patients do not distribute homogeneously in asymmetric gonads. 45,X lines predominate over 46,XY in streak gonads, while the relationship between these two is more equivalent in dysgenetic testes, suggesting that testicular or streak differentiation is related to the balance between X0 and XY lines. Testicular dys-genesis is more severe when there is a frank predominance of X0 or XX cells. Higher percentages of XY cells coincide with lesser degrees of dysgenesis. DNA densitometry indicate a higher incidence of neoplastic transformation than previously anticipated. Various specimens showed clear aneuploid histograms but no clear indication of a cytological CIS phenotype. There was a wide cytological variation in aneuploid germ cells, ranging from normally looking big infantile spermatogonia to gonocyte/CIS cells. Aneuploidy probably precedes the full expression of the CIS phenotype. In case of doubt we recommend DNA densitometry to either confirm or discard their neoplastic nature. The earliest recognizable change in germ cell tumorigenesis is probably the polyploidisation of fetal germ cells, followed by the expression of the CIS phenotype in isolated germ cells scattered along infantile seminiferous tubules that later proliferate to give an adult type CIS pattern. PMID- 12760351 TI - Bibliography of food microbiology. PMID- 12760350 TI - Bibliography. Current world Literature. Clinical therapeutics. PMID- 12760353 TI - Bibliography. Current World Literature. Rheumatoid arthritis. PMID- 12760354 TI - Bibliography. Current World Literature. Crystal deposition diseases. PMID- 12760352 TI - [Antimicrobials in primary care. Scenario and identification of problems]. PMID- 12760355 TI - [Current status of antibiotic resistance in three principle species causing infection in primary practice]. PMID- 12760356 TI - [Antimicrobial resistance in primary care. Social factors]. PMID- 12760357 TI - [Antimicrobial resistance in primary care. Correct measures in microbiology]. PMID- 12760358 TI - [Strategies to break antibiotic resistance of respiratory pathogens in primary care]. PMID- 12760359 TI - [New antibiotics in primary care]. PMID- 12760360 TI - [Antimicrobials. Is etiologic treatment possible in primary care?]. PMID- 12760361 TI - [Empirical treatment or evidence based treatment]. PMID- 12760362 TI - [Antimicrobial consumption. Information sources and unity of measures]. PMID- 12760363 TI - [Classic antibiotics in primary care]. PMID- 12760364 TI - International childhood cancer day. All clear for acrylamide? PMID- 12760365 TI - International childhood cancer day. Urgent need for palliative care expertise in India. PMID- 12760366 TI - International childhood cancer day. Melanoma: an unexpected decrease in deaths. PMID- 12760367 TI - Tumor vascularity and tryptase-positive mast cells correlate with a poor prognosis in melanoma. AB - BACKGROUND: The importance of angiogenesis in melanoma has been controversial and is not homogeneous. Mast cell density (MCD) is highly correlated with the extent of both normal and pathological angiogenesis, such as that in chronic inflammatory diseases and tumours. METHODS: We evaluated the prognostic significance of tumour microvascular density (MVD) and MCD in 25 advanced melanoma patients after resection and a 4-5-year follow up: 48% of the patients were alive and free of metastases (good prognostic subgroup); 16% had developed regional nodal metastases (intermediate prognostic subgroup); and 36% had died (poor prognostic subgroup). Tissues samples were investigated immunohistochemically to count microvessels and mast cells with an antifactor VIII and an antitryptase antibody, respectively. RESULTS: Immunohistological staining showed a higher number of microvessels and mast cells in melanoma lesions of poor prognosis as compared with intermediate prognosis and with good prognosis, respectively. CONCLUSIONS: These data agree with those showing a close relationship between MCD and angiogenesis during tumour progression and demonstrate, for the first time, a prognostic significance of MCD in human melanoma. PMID- 12760369 TI - From cell biology to biotechnology in space. AB - In this article I discuss the main results of our research in space biology from the simple early investigations with human lymphocytes in the early eighties until the projects in tissue engineering of the next decade on the international space station ISS. The discovery that T lymphocyte activation is nearly totally depressed in vitro in 0 g conditions showed that mammalian single cells are sensitive to the gravitational environment. Such finding had important implications in basic research, medicine and biotechnology. Low gravity can be used as a tool to investigate complicated and still obscure biological process from a new perspective not available to earth-bound laboratories. Low gravity may also favor certain bioprocesses involving the growth of tissues and thus lead to commercial and medical applications. However, shortage of crew time and of other resources, lack of sophisticated instrumentation, safety constraints pose serious limits to biological endeavors in space laboratories. PMID- 12760370 TI - Rite of passage? Why young adults become uninsured and how new policies can help. PMID- 12760371 TI - Plant cells on earth and in space. AB - Two quite different types of plant cells are analysed with regard to transduction of the gravity stimulus: (i) Unicellular rhizoids and protonemata of characean green algae; these are tube-like, tip-growing cells which respond to the direction of gravity. (ii) Columella cells located in the center of the root cap of higher plants; these cells (statocytes) perceive gravity. The two cell types contain heavy particles or organelles (statoliths) which sediment in the field of gravity, thereby inducing the graviresponse. Both cell types were studied under microgravity conditions (10(-4) g) in sounding rockets or spacelabs. From video microscopy of living Chara cells and different experiments with both cell types it was concluded that the position of statoliths depends on the balance of two forces, i.e. the gravitational force and the counteracting force mediated by actin microfilaments. The actomyosin system may be the missing link between the gravity-dependent movement of statoliths and the gravity receptor(s); it may also function as an amplifier. PMID- 12760372 TI - Effects of microgravity on vestibular development and function in rats: genetics and environment. AB - Our anatomical and behavioral studies of embryonic rats that developed in microgravity suggest that the vestibular sensory system, like the visual system, has genetically mediated processes of development that establish crude connections between the periphery and the brain. Environmental stimuli also regulate connection formation including terminal branch formation and fine-tuning of synaptic contacts. Axons of vestibular sensory neurons from gravistatic as well as linear acceleration receptors reach their targets in both microgravity and normal gravity, suggesting that this is a genetically regulated component of development. However, microgravity exposure delays the development of terminal branches and synapses in gravistatic but not linear acceleration-sensitive neurons and also produces behavioral changes. These latter changes reflect environmentally controlled processes of development. PMID- 12760373 TI - Anuran metamorphosis: a model for gravitational study on motor development. AB - Limbs and supporting structures of an organism experience a full weight of its own when it lands from water, because neutral buoyancy in the aquatic habitat will be no longer available in the terrestrial world. Metamorphosis of anuran amphibians presents a good research model to examine how this transition from non loading to weight-loading affects development of motor capacity at the time of their first emergence on land. Our video analysis of the transitional anurans, Rana catesbeiana, at Gosner stage 46 (the stage of complete transformation) demonstrated that the take-off speed increased 1.23-fold after the first six hours of weight-loading on the wet ground. It did not increase further during the following three days of loading, and was close to the level of mature frogs with different body mass. During development of larvae in deep water with no chance of landing through metamorphosis, both tension and power of a hindlimb anti-gravity muscle increased 5-fold between stages 37 and 46. However, the muscle contractility increased more rapidly when the larvae could access the wet ground by their natural landing behavior after stages 41-42. Muscle power, one of major factors affecting locomotory speed, was 1.29-fold greater in the loaded than in the non-loaded larvae at the transitional stage. Thus, weight-loading had a potentially significant effect on the elevation of motor capacity, with a similar extent of increment in locomotory speed and muscle power during the last stages of metamorphosis. Such a motor adjustment of the froglets in a relatively short transitional period would be important for effective ecological interactions and survival in their inexperienced terrestrial life. PMID- 12760374 TI - Effects of microgravity and hypergravity on aging and longevity of insects. AB - The effects of microgravity and hypergravity on aging are still poorly documented, particularly in mammals. However, there is a growing interest for the use of the fruit fly, Drosophila melanogaster, and this species may be now considered as a model organism in gravitational biology studies dealing with aging. PMID- 12760376 TI - Victorian children and sex: the reality ignored by proponents of child sexual rights. PMID- 12760375 TI - Future opportunities for life science programs in space. AB - Most space-related life science programs are expensive and time-consuming, requiring international cooperation and resources with trans-disciplinary expertise. A comprehensive future program in "life sciences in space" needs, therefore, well-defined research goals and strategies as well as a sound ground based program. The first half of this review will describe four key aspects such as the environment in space, previous accomplishments in space (primarily focusing on amphibian embryogenesis), available resources, and recent advances in bioinformatics and biotechnology, whose clear understanding is imperative for defining future directions. The second half of this review will focus on a broad range of interdisciplinary research opportunities currently supported by the National Aeronautics and Space Administration (NASA), National Institute of Health (NIH), and National Science Foundation (NSF). By listing numerous research topics such as alterations in a diffusion-limited metabolic process, bone loss and skeletal muscle weakness of astronauts, behavioral and cognitive ability in space, life in extreme environment, etc., we will attempt to suggest future opportunities. PMID- 12760377 TI - Genetic engineering to study testicular tumorigenesis. AB - In humans, Sertoli cell tumors account for approximately 4% of all testicular tumors, and 20% of these are malignant. The mechanisms underlying Sertoli cell tumorigenesis remain largely unknown. Using gene knockout technology, we previously generated mutant mice lacking the alpha subunit of inhibin dimers. The inhibin alpha-null male mice develop testicular Sertoli cell tumors with 100% penetrance. These tumors develop as early as 4 weeks of age and cause a cachexia like wasting syndrome. Castrated inhibin alpha knockout mice develop sex steroidogenic adrenal cortical tumors. These studies have identified inhibins as secreted tumor suppressors with specificity for the gonads and adrenal glands. It had been suggested that endocrine factors play roles in Sertoli cell tumorigenesis by altering cell cycle machinery of the Sertoli cells. To test the potential of these factors to function as modifiers of Sertoli cell tumorigenesis, we have employed a genetic intercross strategy, breeding inhibin a mutant mice with mutant mice deficient in endocrine signaling factors including gonadotropin releasing hormone (hypogonadal, hpg mice), follicle stimulating hormone, anti-Miillerian hormone (AMH), activin receptor type II, or androgen receptor (testicular feminization, tfm mice), or mice overexpressing follistatin. We are also investigating the effects of loss of critical cell cycle regulators, such as cyclin dependent kinase inhibitor p27, on Sertoli cell tumorigenesis in inhibin alpha knockout males. These studies clearly demonstrate the roles of these factors as modifiers of the Sertoli cell tumorigenesis. Activin signaling through activin receptor type II is responsible for the cachexia-like syndrome observed in the inhibin a knockout mice with tumors. The gonadotropin hormones are essential for testicular tumor development, but elevated FSH levels are not sufficient to cause Sertoli cell tumors. Absence of FSH, lack of androgen receptor, or overexpression of follistatin slows the tumor growth and minimizes the cachexia symptoms, thus prolonging the life span of these double mutant mice. In contrast, absence of AMH or p27 causes earlier onset and more aggressive development of testicular tumor, with an earlier death of double mutant mice. We are currently investigating roles of estrogen signaling pathways, and other cell cycle regulators, in tumor development in the inhibin alpha knockout mice by generating mice with double or triple mutations. Genetic engineering in mouse models provides a powerful tool to study the mechanisms of testicular tumorigenesis and define the important genetic modifiers in vivo. PMID- 12760378 TI - Expression of Wnt and Notch pathway genes in a pluripotent human embryonal carcinoma cell line and embryonic stem cell. AB - Embryonal carcinoma (EC) cells, the pluripotent stem cells of teratocarcinomas, show many similar-ities to embryonic stem (ES) cells. Since EC cells are malignant but their terminally differentiated derivatives are not, understanding the molecular mechanisms that regulate their differentiation maybe of value for diagnostic and therapeutic purposes. We have examined the expression of multiple components of two developmentally important cell-cell signalling pathways, Wnt and Notch, in the pluripotent human EC cell line, NTERA2, and the human ES cell line, H7. Both pathways have well-documented roles in controlling neurogenesis, a process that occurs largely in response to retinoicacid (RA) treatment of NTERA2 cultures and spontaneously in H7 cultures. In NTERA2, many ofthe genes tested showed altered transcriptional regulation following treatment with RA. These include members of the frizzled gene family (FZDI, FZD3, FZD4, FZD5, FZD6), encoding receptors forWnt proteins, the Frizzled Related Protein family (SFRPI, SFRP2, FRZB, SFRP4), encoding solubleWnt antagonists and also ligands and receptors of the Notch pathway (Dlkl, Jaggedl; Notchl, Notch2, Notch3). Few differences were found in the repertoire of Wnt and Notch pathway genes expressed by NTERA2 EC cells and H7 ES cells. We present a model in which interactions between and regulation of Wnt and Notch signalling are important in maintaining EC/ES stem cells and also controlling their differentiation. PMID- 12760380 TI - Visual field defects--essentials for neurologists. PMID- 12760379 TI - Deregulation of the G1/S-phase control in human testicular germ cell tumours. AB - Deregulated cell cycle and defective genome-integrity checkpoints are among the hallmarks of cancer. Here we summarize our recent studies of key components of the GI/S machinery in normal human spermatogenesis, and their abnormalities in testicular germ cell tumours (TGCTs), with special emphasis on carcinoma in situ lesions (CIS). Our combined immunohistochemical and immunoblotting analyses of normal human adult and fetal testes, CIS, seminomas, embryonal carcinomas, and teratomas, revealed an 'unorthodox' spectrum of defects within the so-called RB pathway in TGCTs. The early aberrations included lack of expression of the retinoblastoma tumour suppressor (pRB) and the CDK inhibitor pl9ink4d, and overexpression of cyclin D2. Progression from CIS to invasive TGCTswas associated with loss of another two CDK inhibitors and tumour suppressors: pl6ink4a and pl8ink4c. We also found the lack of pRB and pl9ink4d in fetal gonocytes, the candidate target cell for all types of TGCTs. These findings, together with the status of the Chk2-p53 DNA-integrity checkpoint, are considered in relation to the origin, biology and pathogenesis of TGCTs, and potential implications of the GI/S defects for the curability of these tumours. PMID- 12760381 TI - [Past and future studies of memory]. PMID- 12760382 TI - The tremor associated with non-paraproteinaemic acquired demyelinating polyneuropathy--a case study. PMID- 12760383 TI - The stiff limb syndrome--a new case and a literature review. PMID- 12760384 TI - Clinical and radiological mimicry of vCJD in a valine homozygous PrP(Sc) type 1 sCJD patient. PMID- 12760385 TI - Bilateral stroke secondary to pituitary apoplexy. PMID- 12760386 TI - Progressive myelopathy with selective involvement of the lateral and posterior columns after inhalation of heroin vapour. PMID- 12760387 TI - Recommendations for reporting weight loss. PMID- 12760388 TI - Restless Legs Syndrome induced by impairment of sensory spinal pathways. PMID- 12760389 TI - Vertebral angioplasty for treatment of transient monocular blindness. PMID- 12760390 TI - The role of the thoracic surgeon in minimally invasive esophageal surgery: past, present, and future. PMID- 12760391 TI - Central sleep apnoea and unilateral diaphragmatic paralysis associated with vertebral artery compression of the medulla oblongata. PMID- 12760392 TI - Unilateral palsy of all cranial nerves (Garcin syndrome) in a patient with rhinocerebral mucormycosis. PMID- 12760393 TI - Hypergraphia, verbal aspontaneity and post-stroke depression secondary to right cyngulate and corpus callosum infarction. PMID- 12760394 TI - Pontine pseudoathetosis: lemniscal involvement visualized by by axonal tracking method with diffusion tensor imaging. PMID- 12760395 TI - Otfrid Foerster (1873-1941). PMID- 12760397 TI - Test your knowledge. Chronic dermatitis. PMID- 12760396 TI - Hit-Hit and hit-Run: viruses in the playing field of multiple sclerosis. AB - Viruses have been major players in the search for the cause of multiple sclerosis (MS). In support of the viral theory is the predominance of CD8+ T cells and class-I major histocompatibility complex in lesions, the powerful therapeutic effects of beta interferons, the ease of inducing demyelination in experimental models following virus challenge, and the documented examples of several human demyelinating diseases conclusively demonstrated to be of viral origin. We propose two hypotheses of how viruses may cause MS. In the "Hit-Hit" hypothesis, the virus persists or may be reactivated in the central nervous system (CNS). Injury is the result of direct viral damage and by an attempt of the immune response to clear the infectious agent. In the "Hit-Run" hypothesis, virus infects the periphery but never enters the CNS. The virus sets up an abnormal immunologic milieu for subsequent autoimmunity. In both scenarios, knowing the inciting virus would be expected to eliminate disease if the population were vaccinated to prevent infection. In the treatment of patients with fully established disease, the Hit-Hit hypothesis would require that antiviral agents enter the CNS and stop replication. In the case of the Hit-Run hypothesis, treatment of patients with established disease with antiviral agents would be futile. PMID- 12760398 TI - Regression of hepatic fibrosis and cirrhosis in patients with chronic hepatitis C treated with interferon-based therapy. PMID- 12760399 TI - Problem of imbalance in risk factors in clinical trials: logistic regression is the answer. PMID- 12760400 TI - Phenobarbital-induced alterations in the sexual differentiation of the female rat: reversal by hydroxyurea and cycloheximide. AB - The possible mechanism of action of phenobarbital in prenatal female rats has been studied using different inhibitors of protein/DNA/RNA synthesis. Administration of phenobarbital (40 mg/kg/day) subcutaneously from day 17 to day 20 of pregnancy resulted in a delay of the onset of puberty, disorders of estrous cycle, infertility, and high estrogen levels in the female offspring. Cycloheximide (60 microg/kg/day)--a protein synthesis inhibitor and hydroxyurea (160 mg/kg/day)--a DNA synthesis inhibitor when combined with phenobarbital, significantly reduced the incidence of reproductive dysfunctions listed above. Alpha-amanitin (15 microg/kg/day), an inhibitor of m-RNA synthesis, produced reproductive disorders in the female offspring when administered alone to pregnant rats and also failed to reverse the PB-induced reproductive alterations. Cycloheximide and hydroxyurea produced no effect upon reproductive function when administered alone. Cycloheximide administration resulted in low body weight at birth and delayed eye opening. These results suggest that prenatal administration of phenobarbital produces its effect via new protein synthesis and that cycloheximide and hydroxyurea protect the offspring from phenobarbital-induced toxic effects by inhibiting its biochemical action. PMID- 12760401 TI - Chloramphenicol succinate kinetics in infants and young children. AB - We sought to estimate the serum and urine pharmacokinetics of chloramphenicol succinate (CmS) and the effects of variation of these parameters on chloramphenicol (Cm) kinetics in 24 infants and young children ages two weeks to seven years. The mean T(1/2) of CmS was 0.40 hours; the mean body clearance was 0.72 liter/KG/hour; the mean apparent volume of distribution was 0.42 liter/kg. Variation in CmS T(1/2) did not correlate with significant variation in Cm T(1/2) (r2 = 0.002, P = 0.84). Urine collected during the dosing interval in nine patients contained 35% (mean) of the administered dose. Adjusting the infusion duration to 5 minutes or 120 minutes had no effect on the amount of CmS lost in the urine. The quantity of CmS lost in the urine affects the amount bioavailable, and secondarily the calculated volume of distribution and body clearance of Cm. We conclude that variation in urinary prodrug excretion affects the amount of Cm bioavailable to the patient, but variation in CmS T(1/2) has little effect on Cm T(1/2). PMID- 12760402 TI - The blocking effect of bupivacaine on the laryngeal chemoreflex and the trigeminal diving reflex in newborn lambs. AB - This is a study of the laryngeal chemoreflex (LCR) and the trigeminal diving reflex (TDR) responses in the unanesthetized newborn lamb and of the changes in these responses produced by administration of intravenous bupivacaine. It was determined that both 1 mg and 10 mg bupivacaine significantly reduced the LCR. The responses to water and saline stimulation were similar after bupivacaine suggesting a complete blocking effect on the chemoreceptors. When the LCR was initiated by electrical stimulation of the cut superior laryngeal nerve the response was not reduced by bupivacaine indicating that the most likely site of action of bupivacaine is the chemoreceptors in the laryngeal mucosa. The trigeminal diving reflex was significantly reduced after 10 mg bupivacaine, whereas 1 mg was ineffective. PMID- 12760403 TI - Prenatal methadone exposure: effects on behavior in early infancy. AB - As part of an ongoing longitudinal study of the developmental effects of prenatal methadone exposure, 41 children born to methadone-maintained mothers and 23 children from matched backgrounds but with negative maternal history of drug abuse were evaluated at six months of age. Each child received physical and neurological examinations and a battery of behavioral assessments that included a visual habituation task, the Bayley Scales and the Object Permanence Scales. The groups did not differ significantly in frequency of suspect-abnormal neurological signs or in mean scores on the three behavioral measures. Despite the great within-group variances, performance on the behavioral measures was not related to maternal or neonatal characteristics. There were significantly more low PDI scores (predictors of developmental difficulties) among methadone subjects, particularly among methadone vs comparison males. These findings corroborate other studies that have shown 1) delayed motor development in methadone-exposed infants, 2) greater vulnerability of males to adverse environmental conditions, and 3) correlation between early methadone exposure and behavioral abnormalities in adult male rats. The significance of prenatal methadone exposure as a risk factor is discussed. PMID- 12760404 TI - Recommended amikacin doses in newborns often produce excessive serum levels. AB - Emergence of a multiply drug resistant Enterobacter cloacae during a seven-week period in 1980 caused amikacin to become the aminoglycoside of choice in the initial management of suspected sepsis in a neonatal intensive care unit. Recommended doses (7.5-10 mg/kg loading; 15 mg/kg in two divided doses IV) were given to 5 infants < or = 1,000 gm and to 13 larger babies. Trough levels 11.5 hours after a dose were 16.6 +/- 11.9 microg/ml in infants < or = 1,000 gm and 6.5 +/- 4.3 microg/ml in the larger infants (P < 0.02). Peak levels one hour postinfusion exceeded 40 microg/ml in 3 of 5 < or = 1,000-gm babies and 4 of 12 > 1,000-gm infants (P = NS). Overall, 7 of 10 peak and/or trough levels in < or = 1,000-gm infants were in the range considered toxic in adults, versus 7 of 24 in larger babies (P = 0.03). These data show that surprisingly excessive blood levels of amikacin are likely in infants < or = 1,000 gm and may also occur in larger infants using currently recommended dosage schedules. These unexpected findings emphasize the need to monitor drug levels and individualize therapy in very low birthweight infants. PMID- 12760405 TI - Recovery of prostaglandin production associated with reopening of the ductus arteriosus after indomethacin treatment in preterm infants with respiratory distress syndrome. AB - Increased PGE production has been demonstrated in 9 of 17 preterm infants with patent ductus arteriosus (PDA) associated with respiratory distress syndrome (RDS). Inhibition of PGE production in eight preterm infants with PDA and RDS was associated with marked improvement in the respiratory and circulatory function of all of them. However, in six of them this effect was only transient. In the posttreatment period of five and a half days reopening of the ductus arteriosus was frequently associated with increased PGE production and a drop of indomethacin serum levels. Three of these six infants were transferred for surgical ligation whereas the other three were successfully treated with a second course of indomethacin. However, the margin between closure of the ductus arteriosus and the deterioration of kidney function in preterm infants treated with a presently recommended indomethacin dosage was narrow. In conclusion, until an acceptable therapeutic serum level of indomethacin for ductal closure in preterm infants has been established and the duration of effective prostaglandin synthesis inhibition is known, it is too early for a general recommendation of a dosage regime of indomethacin for the pharmacological closure of PDA in infants with RDS. PMID- 12760407 TI - Effect of age and sex on theophylline clearance in young subjects. AB - The joint effects of age and gender on theophylline plasma clearance (Clp) were assessed using an analysis of covariance technique. Data from several literature sources and from our laboratory were collated for this analysis with only non smoking subjects aged 1.3 to 30.0 years considered. The values of Clp exhibited markedly greater variability in the younger subjects, necessitating development of a weighted linear least-squares program to fit the data. The data exhibited a linear decrease in Clp with increasing age regardless of sex. A gender influence was evident over the age range of 4 to 20 years. Male Clp values tended to be higher than those of females over this age interval [25.5 ml/hr/kg higher (31%) at 4 years and 12.5 ml/hr/kg higher (22%) at 20 years]. These differences are statistically significant, and since they represent a divergence of greater than 20%, may be clinically significant as well. The statistical technique employed may be of value in future pharmacokinetic studies for the examination of multiple effects on pharmacokinetic parameters. PMID- 12760406 TI - Tobramycin and gentamicin are equally safe for neonates: results of a double blind randomized trial with quantitative assessment of renal function. AB - The frequent use of aminoglycosides in neonates led us to compare tobramycin and gentamicin because some studies in adults and animals suggested a safety advantage for tobramycin. In a randomized, double-blind comparison, 50 infants < or = 3 days old received either tobramycin or gentamicin, 2.5 mg/kg/12 hr, for a mean of 8 days. The groups were similar for gestational age, birth weight, underlying disease, Apgar score, clinical condition, renal function, and concurrent use of other drugs. Trough and postdose serum concentrations of both drugs were similar in each weight group. Renal status at onset, during or up to 2 wk after therapy was also similar, as judged by urinalysis, serum creatinine, fractional excretion of Na, urine N-acetyl-beta-glucosaminidase, and urine to serum creatinine ratio. Nephrotoxicity occurred in four infants (13%) receiving tobramycin and three infants (15%) receiving gentamicin. No otoxicity, as measured by auditory brainstem response, was noted in either group. Tobramycin offered no safety advantages over gentamicin in neonates, and the choice between the agents should be based on other considerations, such as susceptibility of the pathogen and cost. PMID- 12760409 TI - The influence of rotational exercises on freezing in Parkinson's disease. AB - The advanced stage of Parkinson's disease (PD) is characterised by complex movement disturbances including freezing. Because freezing is resistant to drug therapy, there has recently been renewed interest in non-pharmacological treatment programmes. In this study the effect of rotational stimulation on freezing was investigated. Eight patients with idiopathic PD and freezing participated in the study. Switching from foot-lifting to 'stand up and walk' provoked freezing in all the tested patients. The mean OPMSP (Onset of Premotor Silence Period) in the EMG of the m. tibialis anterior during the 'foot-lifting' sessions preceding freezing was 124 msec. This value, together with a striking repetition of the EMG discharges in the m. tibialis anterior following the request to 'stand up and walk', was related to the inducement of freezing, as a decrease of 42 msec in the OPMSP (t = 2.61; p < or = 0.01) after rotational stimulation abolished freezing and the concomitant EMG disturbances. Rotational stimulation also reduced freezing frequency during daily life. Freezing periods fell to below 50% of the pre-treatment level the day following rotation (t = 5.58; p < or = 0.001). The OPMSP predicted the short and long-term effects of rotational exercises (R = 0.74; p < or = 0.03). The rather long-lasting effect of the stimulation suggests a possible modulation of neurochemical transmission. Further studies are required to shed more light on the point of action and to elucidate which neurotransmitter might be involved. PMID- 12760408 TI - TorsinA, microtubules and cell polarity. AB - Early-onset primary dystonia is an inherited disorder characterized by involuntary twisting, repetitive movements and abnormal postures. It has recently been demonstrated that the DYT1 gene is the most relevant gene associated with primary generalized dystonia. The DYT1 gene product is a 332-aminoacid long protein, termed TorsinA, whose function is still not clear. Based on the results obtained in other species, we proposed that TorsinA, similarly to OOC-5 in nematodes, directs and/or stabilizes the subcellular localization of specific kinases, which may in turn phosphorylate microtubule associated proteins, such as tau. In this way, TorsinA may contribute to maintaining the appropriate site directed polarization and control neurite outgrowth. PMID- 12760410 TI - Headache and the lower cervical spine: long-term, postoperative follow-up after decompressive neck surgery. AB - A retrospective search for headache sufferers was conducted among patients operated on for cervicobrachialgia, and the operative results were evaluated. We also tried to classify the preoperative headache according to current headache classification systems. A total of 187 patients were operated on with Smith Robinson's method, or by "simple" foraminectomy/ facetectomy. Headache was present in 23, and 17 of these could be followed up for an average post-operative observation period of 8.5 years. The operation seemed to result in headache improvement in 15 patients, i.e., excellent in 7; good in 2; moderate in 6; in other words: "excellent"/"good" in 53%. Improvement of neck pain was found in 15 patients (excellent in 5; good in 4; moderate in 6). The headache characteristics were generally compatible with the criteria for cervicogenic headache (CEH) (1990 version). CEH can probably stem from a pathology in the lower cervical spine. This should probably be taken into account when evaluating the individual patient with symptoms reminiscent of CEH. This headache may benefit from operations directed towards the cervical spine. PMID- 12760411 TI - Preprogramming motor dysfunction in paroxysmal kinesigenic choreoathetosis. AB - Paroxysmal kinesigenic choreoathetosis (PKC) is characterized by abnormal involuntary movements precipitated by sudden movement. As a result, a possible impairment of cerebral organization of voluntary motor activity is hypothesized in PKC. We examined a 14-year-old boy affected by a sporadic form of PKC, adopting a multimodal psychophysiological approach, including P300, contingent negative variation (CNV) and a specific paradigm for the study of movement related potentials (MRPs). Recordings were made before and after phenobarbital therapy. No changes were observed in the non-motor parameters (P300 and early wave of the CNV), whereas the premotor CNV component and the electrophysiological components, reflecting the preprogramming activity of a voluntary motor act, showed selective modifications induced by the anticonvulsant therapy. Our PKC patient presents a disorder of temporal organization of a voluntary motor response to a stimulus. Both a clinical improvement and normalization of motor related electrophysiological anomalies were observed during phenobarbital (PB) therapy. PMID- 12760412 TI - Bit-mapped quantitative EEG analysis in a tiger (Tiger felis) with partial seizures: a case report. AB - We report electroencephalographic findings in an anesthetized 4-month-old female drug-naive tiger (Tiger felis) affected by partial seizures with secondary generalization. Both clinical signs and electroencephalographic abnormalities were consistent with a forebrain lesion. Recurring epileptiform activity was noted in the left frontal, central and temporal derivations upon visual inspection of the electroencephalogram (EEG). A quantified EEG, displayed on brain maps, showed the predominance of delta and theta slow rhythms. As regards the absolute power, a prevalence of left frontal-temporal activity was noted. An infectious or inflammatory condition was thought to be the most probable cause of the symptomatic epilepsy in our patient. Unfortunately, other differential diagnoses could not be ruled out. PMID- 12760413 TI - Memory disturbances and temporal lobe epilepsy simulating Alzheimer's disease: a case report. AB - We report the case of a patient presenting with recurrent transient dyscognitive episodes and memory disturbances, simulating the clinical picture of early Alzheimer's disease (AD). Neuropsychological examination showed only episodic memory impairment without significant progression over time in the absence of any other cognitive dysfunctions; magnetic resonance imaging failed to show selective temporal atrophy. The suspicion that a nocturnal epileptic seizure had occurred arose on the basis of a clinical report of tongue biting and the findings of sharp waves over the temporal region on standard EEG. Thus 600 mg/day of carbamazepine was added to donezepil therapy. Antiepileptic treatment completely reversed the cognitive disturbances. Our observation stresses the possibility that unrecognised epilepsy may present as early AD. PMID- 12760415 TI - Palindromes in proteins. AB - Palindromes in DNA consist of nucleotides sequences that read the same from the 5'-end to the 3'-end, and its double helix is related by twofold axis. They occur in genomes of all organisms and have various functions. For example, restriction enzymes often recognize palindromic sequences of DNA. Palindromes in telomeres are crucial for initiation of replication. One can ask the questions, Do palindromes occur in protein, and if so, what function they play? We have searched the protein SWISSPROT database for palindromic sequences. A great number (26%) of different protein palindromes were found. One example of such protein is systemin, an 18-amino-acid-long peptide. It contains palindrome in its beta-sheet domain that interacts with palindromic fragment of DNA. The other palindrome containing protein is cellular human tumor suppressor p53. Oligonucleotide LTI ITL has been observed in the crystal structure and is located close to a DNA recognizing domain. As the number of possible palindromic sequences of a given length is far much greater for proteins (20N) than for nucleic acids (4N), the study on their role seems to be an exciting challenge. Our results have clearly showed that palindromes are frequently occurring motives in proteins. Moreover, even very few examples that we have examined so far indicate the importance of further studies on protein palindromes. PMID- 12760414 TI - Current insights into familial spastic paraparesis: new advances in an old disease. AB - Hereditary spastic paraparesis (HSP) comprises a clinically and genetically heterogeneous group of disorders characterized by progressive spasticity and hyperreflexia of the lower limbs. The past few years have witnessed an exponential increase in knowledge of this disease and we can now list 19 loci mapped on the human genome and eight genes cloned. However, this wider knowledge of the molecular basis of HSP has had limited impact on clinical practice: the use of antispastic drugs and regular physiotherapy still remain crucial in the therapeutic management of patients. Nonetheless, the identification of new genes mutated in HSP furthers comprehension of the pathomechanisms involved and helps in genetic counseling, especially of asymptomatic individuals who request molecular analyses. PMID- 12760416 TI - Reversible aggregation of mouse prion protein derivatives with PrPSC-like structural properties. AB - Three carbamylated derivatives of reduced mouse prion protein (mPrP) were isolated during the aborted oxidative folding in the presence of urea. These three prion protein derivatives (mPrP-a, mPrP-b, and mPrP-c) exist as monomer in the acidic solution (pH < 2.0) and exhibit prevalent random coil structure. However, they undergo rapid aggregation and transformation to a predominant beta sheet structure upon exposure to ionic buffer with pH greater than 3.0. The stability of aggregates of mPrP conformers is in part dependent upon the time that they were allowed to develop. The nascent aggregates comprise a significant fraction of loosely packed mPrP monomers that can be dissociated by treatment with strong acidic solution. Matured aggregates acquired through prolonged sample incubation contain more tightly packed mPrP monomers that cannot be dissociated by strong acid but can be disaggregated by denaturant. The properties of reversible aggregation of mPrP-a, mPrP-b, and mPrP-c bear a striking resemblance to that observed with aggregates of hamster PrPSC. PMID- 12760417 TI - Hieronymain I, a new cysteine peptidase isolated from unripe fruits of Bromelia hieronymi Mez (Bromeliaceae). AB - A new peptidase, named hieronymain I, was purified to homogeneity from unripe fruits of Bromelia hieronymi Mez (Bromeliaceae) by acetone fractionation followed by cation exchange chromatography (FPLC) on CM-Sepharose FF. Homogeneity of the enzyme was confirmed by mass spectroscopy (MALDI-TOF), isoelectric focusing, and SDS-PAGE. Hieronymain is a basic peptidase (pI > 9.3) and its molecular mass was 24,066 Da. Maximum proteolytic activity on casein (>90% of maximum activity) was achieved at pH 8.5-9.5. The enzyme was completely inhibited by E-64 and iodoacetic acid and activated by the addition of cysteine; these results strongly suggest that the isolated protease should be included within the cysteine group. The N-terminal sequence of hieronymain (ALPESIDWRAKGAVTEVKRQDG) was compared with 25 plant cysteine proteases that showed more than 50% of identity. PMID- 12760418 TI - Purification and characterization of loxnecrogin, a dermonecrotic toxin from Loxosceles gaucho brown spider venom. AB - The most common manifestation of Loxosceles spider envenoming is a dermonecrotic lesion at the bite site. Dermonecrotic toxins from Loxosceles gaucho venom were purified and characterized by mass spectrometry (capillary liquid chromatography followed by mass spectrometry detection). Two components were purified: a major one of 31,444 Da, called loxnecrogin A, and a minor one of 31,626 Da, called loxnecrogin B, being probably two isoforms of the toxin. The N-terminal sequence of loxnecrogin A showed similarity with N termini of other sphingomyelinolytic dermonecrotic toxins isolated from venoms of different Loxosceles species. The internal sequences did not present any statistically significant hits in sequence databases searches. However, loxnecrogin A partial sequence showed high similarity to regions of L. intermedia LiD1 recombinant protein sequence, recently described in the literature but not yet deposited in databanks. PMID- 12760419 TI - Inhibition of neurotransmitter release by peptides that mimic the N-terminal domain of SNAP-25. AB - Botulinum neurotoxin serotypes A and E (BoNT/A and BoNT/E) block neurotransmitter release by cleaving the 206-amino-acid SNARE protein, SNAP-25. For each BoNT serotype, cleavage of SNAP-25 results in the loss of intact protein, the production of an N-terminal truncated protein, and the generation of a small C terminal peptide. Peptides that mimic the C-terminal fragments of SNAP-25 following BoNT/A or BoNT/E cleavage were shown to depress transmitter release in bovine chromaffin cells and in Aplysia buccal ganglion cells. Similarly, the N terminal-truncated SNAP-25 resulting from BoNT/A or BoNT/E cleavage has been found to inhibit transmitter exocytosis in various systems. With one exception, however, the inhibitory action of truncated SNAP-25 has not been demonstrated at a well-defined cholinergic synapse. The goal of the current study was to determine the level of inhibition of neurotransmitter release by N-terminal BoNT/A- or BoNT/E-truncated SNAP-25 in two different neuronal systems: cholinergically coupled Aplysia neurons and rat hippocampal cell cultures. Both truncated SNAP-25 products inhibited depolarization-dependent glutamate release from hippocampal cultures and depressed synaptic transmission in Aplysia buccal ganglion cells. These results suggest that truncated SNAP-25 can compete with endogenous SNAP-25 for binding with other SNARE proteins involved in transmitter release, thus inhibiting neurotransmitter exocytosis. PMID- 12760420 TI - Isolating substrates for an engineered alpha-lytic protease by phage display. AB - Panning of a substrate phage library with an alpha-lytic protease mutant showed that substrate phage display can be used to isolate sequences with improved protease sensitivity even for proteases of relatively broad specificity. Two panning experiments were performed with an engineered alpha-lytic protease mutant known to have a preference for cleavage after His or Met residues. Both experiments led to the isolation of protease-sensitive phage containing linker sequences in which His and Met residues were enriched compared with the initial library. Despite the relatively hydrophobic substrate binding site of the enzyme, the predominant protease-sensitive sequence isolated from the second library panning had the sequence Asp-Ser-Thr-Met. Kinetic studies showed that this sequence was cleaved up to 4.5-fold faster than rationally designed positive controls. Protease-resistant phage particles were also selected and characterized, with the finding that Gly and Pro appeared frequently at the putative P4 positions, whereas Asp dominated the putative P1 position. PMID- 12760422 TI - Thr94 in bovine myelin basic protein is a second phosphorylation site for 42-kDa mitogen-activated protein kinase (ERK2). AB - Treatment of bovine brain myelin basic protein with 42-kDa mitogen-activated protein kinase [p42 MAPK or extracellular signal-regulated kinase 2 (ERK2)] in the presence of ATP and Mg2+ results in phosphorylation of Thr94 and Thr97. Thr94 is not previously known to be an ERK2 phosphorylation site. Both residues are phosphorylated to about the same extent and are in the highly conserved segment Asn91-Ile-Val-Thr94-Pro-Arg-Thr97-Pro-Pro-Pro-Ser101 MALDI mass spectrometry before and after ERK2 treatment revealed the addition of two phosphate groups to the protein. Tryptic cleavage resulted in a single fragment (positions 91-104) carrying the observed mass increase. Tandem mass spectrometry applied to the tryptic peptide showed that both Thr94 and Thr97 are acceptors of phosphate. A singly phosphorylated species could not be detected. Identification of the ERK2 phosphorylation site Thr94 in bovine myelin basic protein reveals a nontraditional phosphate acceptor position, preceded by three noncharged residues (Asn-Ile-Val). Proline at position -2 or -3 from the phosphorylation site, typical for the recognition sequence of proline-directed kinases, is missing. The results provide information for delineation of a further substrate consensus motif for ERK2 phosphorylation. PMID- 12760421 TI - Polypeptide components of oligomeric legumin-like thiamin-binding protein from buckwheat seeds characterized by partial amino acid sequencing and photoaffinity labeling. AB - Among thiamin-binding proteins that ubiquitously occur in plant seeds, that of common buckwheat became a model of extensive studies of the chemical mechanism of ligand-protein interaction. In this work, the polypeptide components of buckwheat seed thiamin-binding protein (BSTBP) are identified and characterized. We suggest that BSTBP is probably a fraction of major storage 13 S globulin (legumin), has an average molecular mass of 235 kDa and comprises hexamers of 57-kDa and 38-kDa subunits in variable combinations. Each subunit is a pair of disulfide-linked polypeptide chains, 36 kDa plus 24 kDa and two-times 22 kDa, respectively. The N terminal sequences of 22-kDa and 24-kDa components show strict homology with those reported for "basic subunits" of buckwheat legumin. By photoaffinity labeling of BSTBP with 4-azido-2-nitrobenzoylthiamine, it is shown that the 36 kDa chain plays the major role in thiamin binding, but the other chains may also be variably involved. Putative thiamin-binding fragments are identified and sequenced. PMID- 12760423 TI - Structural properties of porcine intestine acylpeptide hydrolase. AB - The acylpeptide hydrolase of porcine intestinal mucosa (pi-APH) is a serine peptidase belonging to the prolyl oligopeptidase family. The enzyme catalyzes the release of N-terminal acylamino acids, especially acetylamino acids, from acetylpeptides. pi-APH is an homotetramer of approximately 300 kDa. We report the loss of the native tetrameric structure of pi-APH upon citraconylation and the process was reversed at acidic pH, indicating that the subunits were noncovalently bound. Determination of free cysteines in combination with peptide mapping suggested the involvement of all cysteines in disulfide bridges. Two structural domains were identified based on the three-dimensional model of pi-APH monomer: a beta-propeller fold in the N-terminal sequence (113-455) and an alpha/beta hydrolase fold corresponding to the C-terminal catalytic domain (469 732). Preferential cleavage sites for limited proteolysis with trypsin occurred within the beta-propeller domain, in agreement with the three-dimensional model. The putative role of this domain in the specificity mechanism of APH enzymes is also discussed. PMID- 12760424 TI - Combinatorial evolution of high-affinity peptides that bind to the Thomsen Friedenreich carcinoma antigen. AB - Thomsen-Friedenreich (TF) antigen occurs on approximately 90% of human carcinomas, is likely involved in carcinoma cell homotypic aggregation, and has clinical value as a prognostic indicator and marker of metastasized cells. Previously, we isolated anti-TF antigen peptides from bacteriophage display libraries. These bound to TF antigen on carcinoma cells but were of low affinity and solubility. We hypothesized that peptide amino acid sequence changes would result in increased affinity and solubility, which would translate into improved carcinoma cell binding and increased inhibition of aggregation. The new peptides were more soluble and exhibited up to fivefold increase in affinity (Kd approximately equal to 60 nM). They bound cultured human breast and prostate carcinoma cells at low concentrations, whereas the earlier peptides did not. Moreover, the new peptides were potent inhibitors of homotypic aggregation. The maturated peptides will have expanded applications in basic studies of the TF antigen and particular utility as clinical carcinoma-targeting agents. PMID- 12760425 TI - Magnetic resonance imaging findings in infantile spasms: etiologic and pathophysiologic aspects. AB - An etiologic evaluation of 86 patients with infantile spasms is presented and the place of cranial magnetic resonance imaging (MRI) findings within this spectrum is discussed. A total of 103 cranial MRIs, performed between 4 and 72 months of age, were analyzed and classified according to the etiologic and pathophysiologic aspects. Ninety-one percent of cases were diagnosed as symptomatic infantile spasms, and hypoxic-ischemic encephalopathy was the primary cause (30%). The most common involvement was thinning of the corpus callosum in 43 patients (50%), followed by dilation of cerebral ventricles in 32 (36%), delayed myelination in 23 (26.7%), lesions of diencephalic deep gray matter in 17 (19.7%), and enlargement of the subarachnoid space in 12 (14%). Thin corpus callosum and diffuse atrophy were changes mainly associated with hypoxic-ischemic encephalopathy, whereas delayed myelination seemed to be independent from a specific etiology. The results showed that cranial MRI may provide considerable information regarding not only the etiology but also the pathophysiology of infantile spasms. PMID- 12760426 TI - Characteristics of prolonged afterdischarges in children with malformations of cortical development. AB - We investigated aberrant cortical excitability in malformations of cortical development From subdural electrodes, we recorded afterdischarges lasting > or = 6 seconds in 12 of 13 patients with malformations of cortical development and 6 of 10 pediatric patients with nonmalformations of cortical development and reviewed amperage thresholds, distribution of afterdischarges, and motor responses. In patients with malformation of cortical development, motor response thresholds were high; afterdischarge and motor response thresholds, which essentially overlapped, inversely correlated with age (P < .01); afterdischarge thresholds declined with age; and 8 patients showed afterdischarges in remote sites. In nonmalformation of cortical development, afterdischarge thresholds did not significantly correlate with age; motor response thresholds tended to decline with age; and 2 patients had remote afterdischarges. Adolescent patients with malformations of cortical development had lower afterdischarge thresholds than adolescents with nonmalformation of cortical development (P < .05). From their high afterdischarge (and motor response) thresholds, we concluded that preadolescent patients with malformation of cortical development have less excitable, immature cortices, whereas adolescent patients with malformation of cortical development with low afterdischarge thresholds have hyperexcitable cortices. Remote afterdischarges over focal dysplastic cortex suggest aberrant cortical excitability and neural circuits. PMID- 12760428 TI - Clinical experience with open-label topiramate use in infants younger than 2 years of age. AB - To assess the efficacy, safety, and tolerability of topiramate in infants younger than 24 months of age, we conducted an open-label, multicenter chart review study of infants who received topiramate. Twenty-eight patients were evaluated. All had refractory epilepsy. The mean age of seizure onset was 3.8 months (range 0-10 months). Refractory infantile spasms were the most common epilepsy syndrome. Among infants without infantile spasms, complex partial seizures were the prominent seizure type in eight, followed by simple partial seizures in six. Topiramate was prescribed as add-on therapy in 25 cases and a s monotherapy in 3 cases. Seven of the eight infantile spasms cases improved on topiramate therapy, attaining topiramate monotherapy in three infants. Half of the infants with other seizure types responded to topiramate. The average treatment duration among topiramate responders was 11 months. Topiramate was prescribed after a mean of 3.3 antiepilepsy drugs had been used in these infants. In no case was topiramate the first prescribed antiepilepsy drug. Adverse effects occurred only in five patients, leading to topiramate discontinuation in two patients. Topiramate was efficacious and well tolerated in infants younger than 24 months of age with refractory epilepsy. Prospective data are needed to corroborate this observation. PMID- 12760427 TI - Oligohydrosis and hyperthermia: pilot study of a novel topiramate adverse effect. AB - A 6-year-old boy with partial complex seizures developed recurrent episodes of hyperthermia 2 months after topiramate was introduced into his antiepilepsy drug regimen. Further investigation revealed that the febrile episodes were related to environmental temperature and physical activity. A pilocarpine iontophoresis sweat test showed that the amount of sweat produced by the child was 5% that of age-matched controls. Topiramate discontinuation resulted in the disappearance of febrile episodes and normalization of sweat quantity in repeat sweat testing. Based on this observation and the previous data on zonisamide and isolated case reports on topiramate-related hyperthermia and the effect on sweat production, topiramate was suspected of causing oligohydrosis. A pilot study was carried out involving 13 additional children and young adults (age range 1-37 years) receiving topiramate. All patients were directly questioned regarding symptoms of decreased sweating and heat intolerance, went through a pilocarpine iontophoresis sweat test, and were compared with 14 age-matched controls who went through the sweat test for unrelated reasons. Nine of the patients were found to have reduced sweat quantity on the pilocarpine iontophoresis sweat test (including index case) (mean 0.089 g/30 minutes, SD 0.082; age-matched control: mean 0.21 g/30 minutes, SD 0.06). Eight of them were children (below 16 years). However, only three patients revealed symptoms related to heat intolerance. Topiramate is most likely responsible for decreased sweat production as detected by a pilocarpine iontophoresis sweat test. The effect seems to be more significant in children than in adults. There is a discrepancy between test results and clinical symptoms. Interestingly, oligohydrosis was found to be a relatively common side effect of zonisamide. Both zonisamide and topiramate share a carbonic anhydrase inhibitor activity. The significance of oligohydrosis in hot climates should not be underestimated. Its extent, the role of sweat test prediction, and clinical significance during topiramate treatment should be further estimated. PMID- 12760429 TI - Speed of performance of children with developmental right hemisphere syndrome and with attention-deficit hyperactivity disorder. AB - Slowness is a common complaint in children with attention-deficit hyperactivity disorder (ADHD) and with developmental right hemisphere syndrome. However, it was our clinical impression that slowness in developmental right hemisphere syndrome was more prominent than in ADHD. Our objective was to assess slowness as operationalized by speed of performance in children with developmental right hemisphere syndrome, children with ADHD, and controls. The research sample comprised 19 children in each group, matched for age, gender, socioeconomic status, IQ, and handedness. The subjects were administered a reaction time battery assessing speed of performance. Overall, the average performance differed among the three study groups (F(2,53) = 2.40, P < .01). Children with developmental right hemisphere syndrome were slower than their peers with ADHD (t(35) = 1.99, P < .05) and slower than controls (t(35) = 4.55, P < .001). Children with ADHD performed more slowly than controls, although for the majority of tasks, this was nonsignificant. We conclude that slowness is an integral and consistent component of developmental right hemisphere syndrome and cannot be attributed only to the ADHD symptomatology. PMID- 12760430 TI - A simple method for diagnosis of autosomal recessive spinal muscular atrophy by denaturing high-performance liquid chromatography. AB - Autosomal recessive spinal muscular atrophy is caused by mutations in the survival motoneuron (SMN) gene. There are two nearly identical copies of this gene present on chromosome 5q13; however, only the telomeric copy of this gene is affected in spinal muscular atrophy. In this study, we describe a new method to detect SMN gene deletion by denaturing high-performance liquid chromatography, which is also simple to perform but is faster and more specific. PMID- 12760431 TI - Childhood epilepsy: what is the evidence for what we think and what we do? AB - This article reviews the strength of the evidence that underlies the current approach to the management of childhood epilepsy. The authors reviewed published, peer-reviewed English literature accessed through PubMed and Cochrane reviews with evidence rated as Class 1 (strongest) to Class 4 (weakest). There is considerable inaccuracy in the diagnosis of seizures and epilepsy syndromes. Sound information supports the consensus that the diagnosis of epilepsy should await two unprovoked seizures. Population-based studies indicate that remission from childhood onset epilepsy occurs in at least 50% of children. It is easier to predict a good seizure outcome than a poor one. Absence of concomitant neurologic handicap and onset before about 12 years of age are the most consistent predictors of remission. Intractability is poorly defined and difficult to predict until several antiepilepsy drugs have been used and failed to control the seizures. Most epilepsy syndrome diagnoses do not yield an accurate prognosis. Social outcome appears unsatisfactory in about 50% of cases without intellectual handicap. Death is rare in childhood epilepsy. Those without severe neurologic handicaps have the same mortality as the general population. We identified only 27 published randomized trials of antiepilepsy drugs in children that compare the efficacy of antiepilepsy drugs, offer treatment of syndromes currently without successful treatment, or have negative effects. There is a pressing need for better definitions of seizures and epilepsy syndromes. The causes of poor social outcome are unclear. Intractability needs a clear definition and randomized trials comparing treatment regimes are sadly lacking. PMID- 12760432 TI - Transient lesion in the splenium of the corpus callosum in acute cerebellitis. AB - We report the reversible magnetic resonance imaging (MRI) findings in a 2-year old girl with acute cerebellitis. The high signal intensity in the cerebellum was obvious on diffusion-weighted images, and the splenium of the corpus callosum also showed high signal intensity on T2-weighted images and on diffusion-weighted images. The lesion in the splenium quickly returned to normal 72 hours later, whereas the hyperintensity in the cerebellum persisted for 1 week until clinical improvement was achieved. The findings in our patient suggested the particular vulnerability of the splenium of the corpus callosum compared to other regions and the importance of the MRI examinations including diffusion-weighted images in the early stage of encephalitis. PMID- 12760433 TI - Central pontine and extrapontine myelinolysis owing to disequilibrium syndrome. AB - Neurologic disorders can be seen in patients with end-stage renal failure owing to complications of hemodialysis or peritoneal dialysis. The disequilibrium syndrome can be seen, usually soon after or toward the end of dialysis. We report a patient with central pontine and extrapontine myelinolysis owing to disequilibrium syndrome. The patient had depressed consciousness, agitation, tremor, stupor and hyperactive deep tendon reflexes toward the end of the second peritoneal dialysis. A brain computed tomographic (CT) scan showed hypodense lesions in pontine and extrapontine locations without radiocontrast medium enhancement After 2 days, the patient had only minimal memory deficits. A control brain CT scan 1 week later showed a decrease of the lesions in central pontine and extrapontine locations. Central pontine and extrapontine myelinolysis should be suspected and investigated in the acute neurologic disorders of dialysis patients. PMID- 12760434 TI - Intracranial calcification with congenital rubella syndrome in a mother with serologic immunity. AB - We report a case of an infant with congenital rubella. The mother had received rubella vaccine at the age of 13 years. Rubella serology was performed on day 34 of pregnancy and the result was interpreted as being a positive titer. The patient was a girl born by cesarean section owing to intrauterine growth retardation and fetal distress after 37 weeks' gestation. A computed tomographic scan at 4 days of age showed several cortical low-density areas and calcifications of the periventricular area and basal ganglia. Magnetic resonance imaging (MRI) performed at 4 weeks of age showed almost similar findings. The infant had serum IgG and IgM antibodies against rubella. Rubella virus ribonucleic acid (RNA) was detected from the serum, urine, and cerebrospinal fluid of the infant. At 2 months of age, the patient showed severe bilateral hearing loss. At 12 months of age, she had mild mental retardation and developmental delay. PMID- 12760435 TI - Neurocysticercosis masquerading as a cerebral infarct. AB - The differential diagnosis of acute focal neurologic deficit in childhood is diverse. We report the case of a child presenting with an acute hemiparesis persisting for longer than 24 hours following a focal seizure. The clinical history, examination findings, and results of cranial magnetic resonance imaging (MRI) were initially interpreted as consistent with an arterial ischemic cerebral infarction. Follow-up cranial MRI performed 9 months later revealed changes indicative of neurocysticercosis. Review of original neuroimaging resulted in a revision of the diagnosis to neurocysticercosis. The clinical history, together with neuroimaging findings, is highly compatible with a diagnosis of neurocysticercosis but unusual because it occurred in a child resident in a nonendemic area who had never traveled to an endemic area and whose diet excluded pork. The case reported raises two important issues. The first is the need to carefully consider the differential diagnosis of acute hemiparesis, including unusual causes. Second, it raises awareness of the potential for neurocysticercosis to occur in low-risk patients in nonendemic areas. PMID- 12760436 TI - Mitochondrial myopathy and respiratory failure associated with a new mutation in the mitochondrial transfer ribonucleic acid glutamic acid gene. AB - We report a novel T14687C mutation in the mitochondrial transfer ribonucleic acid glutamic acid gene in a 16-year-old boy with myopathy and lactic acidosis, retinopathy, and progressive respiratory failure leading to death. A muscle biopsy showed cytochrome c oxidase-negative ragged-red fibers, and biochemical analysis of the respiratory chain enzymes in muscle homogenate revealed complex I and complex IV deficiencies. The mutation, which affects the trinucleotide (TpsiC) loop, was nearly homoplasmic in the muscle DNA of the proband, but it was absent in his blood and in the blood from the asymptomatic mother, suggesting that it may have been a spontaneous somatic mutation in muscle. PMID- 12760437 TI - Left-sided facial nevus with contralateral leptomeningeal angiomatosis in a child with Sturge-Weber syndrome: case report. AB - Sturge-Weber syndrome is characterized by a facial vascular nevus associated with an ipsilateral leptomeningeal angioma. Variants of this classical presentation have been described in the literature, some of which have prognostic significance. We report a magnetic resonance imaging (MRI)-confirmed variant of a leptomeningeal angioma contralateral to the facial nevus. We describe one patient with Sturge-Weber syndrome who presented with a left-sided facial nevus, left eye glaucoma, episodes of left-sided weakness, and right-sided leptomeningeal angiomatosis by gadolinium-enhanced brain MRI. The literature regarding variants of Sturge-Weber syndrome and their prognosis is reviewed. The prognosis for this variant is likely similar to Sturge-Weber syndrome with an ipsilateral leptomeningeal angioma. PMID- 12760438 TI - Do epileptic children treated with valproate have a risk of excessive weight gain? PMID- 12760439 TI - Births, marriages, divorces, and deaths: provisional data for January-March 2002. PMID- 12760440 TI - Are newer drugs better drugs? PMID- 12760441 TI - What diets should we be recommending for obesity? PMID- 12760442 TI - European withdrawal of appetite suppressants. AB - The temporary delicensing of the centrally acting appetite suppressants (first in April 2000, then in May 2001) by the European Commission's Committee on Proprietary Medical Products was premature and possibly unjustified. While their ultimate fate is awaited, the European Drugs Evaluation Agency declined to release the information on which its withdrawal advice was based. PMID- 12760443 TI - Should we recommend low-fat diets for obesity? AB - Dietary fat intake has been blamed for the increase in adiposity and has led to a worldwide effort to decrease the amount of fat in the diet. However, the comparative efficacy of this approach is debatable. Whilst short-term dietary intervention studies show that low-fat diets lead to weight loss in both healthy and overweight individuals, it is less clear if a reduction in fat intake is more efficacious than other dietary restrictions in the long term. The purpose of this systematic review was to determine the effectiveness of low-fat diets in achieving sustained weight loss when used for the express purpose of weight loss in obese or overweight people. A comprehensive search identified six studies that fulfilled our criteria for inclusion (randomized controlled trial, participants either overweight or obese, comparison of a low-fat diet with another type of weight-reducing diet, follow-up period that was at least 6 months in duration and inclusion of participants 18 years or older without serious disease). There were a total of 594 participants in the six trials. The duration of the intervention varied from 3 to 18 months with follow-up from 6 to 18 months. There were no significant differences between low-fat diets and other weight-reducing diets in terms of sustained weight loss. Furthermore, the overall weight loss at the 12-18 month follow-up in all studies was very small (2-4 kg). In overweight or obese individuals who are dieting for the purpose of weight reduction, low-fat diets are as efficacious as other weight-reducing diets for achieving sustained weight loss, but not more so. PMID- 12760444 TI - Effects of sugar intake on body weight: a review. AB - Weight reduction programmes are mainly focused on reducing intake of fat and sugar. In this review we have evaluated whether the replacement of dietary (added) sugar by low-energy sweeteners or complex carbohydrates contributes to weight reduction. In two experimental studies, no short-term differences in weight loss were observed after use of aspartame as compared to sugar in obese subjects following a controlled energy-restricted diet. However, consumption of aspartame was associated with improved weight maintenance after a year. In two short-term studies in which energy intake was not restricted, substitution of sucrose by artificial sweeteners, investigated mostly in beverages, resulted in lower energy intake and lower body weight. Similarly, two short-term studies, comparing the effect of sucrose and starch on weight loss in obese subjects did not find differences when the total energy intake was equal and reduced. An ad libitum diet with complex carbohydrates resulted in lower energy intake compared to high-sugar diets. In two out of three studies, this was reflected in lower body weight in subjects consuming the complex carbohydrate diet. In conclusion, a limited number of relatively short-term studies suggest that replacing (added) sugar by low-energy sweeteners or by complex carbohydrates in an ad libitum diet might result in lower energy intake and reduced body weight. In the long term, this might be beneficial for weight maintenance. However, the number of studies is small and overall conclusions, in particular for the long term, cannot be drawn. PMID- 12760445 TI - How much physical activity is enough to prevent unhealthy weight gain? Outcome of the IASO 1st Stock Conference and consensus statement. AB - A consensus meeting was held in Bangkok, 21-23 May 2002, where experts and young scientists in the field of physical activity, energy expenditure and body-weight regulation discussed the different aspects of physical activity in relation to the emerging problem of obesity worldwide. The following consensus statement was accepted unanimously. 'The current physical activity guideline for adults of 30 minutes of moderate intensity activity daily, preferably all days of the week, is of importance for limiting health risks for a number of chronic diseases including coronary heart disease and diabetes. However for preventing weight gain or regain this guideline is likely to be insufficient for many individuals in the current environment. There is compelling evidence that prevention of weight regain in formerly obese individuals requires 60-90 minutes of moderate intensity activity or lesser amounts of vigorous intensity activity. Although definitive data are lacking, it seems likely that moderate intensity activity of approximately 45 to 60 minutes per day, or 1.7 PAL (Physical Activity Level) is required to prevent the transition to overweight or obesity. For children, even more activity time is recommended. A good approach for many individuals to obtain the recommended level of physical activity is to reduce sedentary behaviour by incorporating more incidental and leisure-time activity into the daily routine. Political action is imperative to effect physical and social environmental changes to enable and encourage physical activity. Settings in which these environmental changes can be implemented include the urban and transportation infrastructure, schools, and workplaces.' PMID- 12760446 TI - Dusting off the epidemiological triad: could it work with obesity? AB - The search for effective ways of dealing with obesity has centred on biological research and clinical management. However, obesity needs to be conceptualized more broadly if the modern pandemic is to be arrested. The epidemiological triad (hosts, agent/vectors and environments) has served us well in dealing with epidemics in the past, and may be worth re-evaluating to this end. Education, behaviour change and clinical practices deal predominantly with the host, although multidisciplinary practices such as shared-care might also be expected to impact on other corners of the triad. Technology deals best with the agent of obesity (energy imbalance) and it's vectors (excessive energy intake and/or inadequate energy expenditure), and policy and social change are needed to cope with the environment. The value of a broad model like this, rather than specific isolated approaches, is that the key players such as legislators, health professionals, governments and industry can see their roles in attenuating and eventually reversing the epidemic. It also highlights the need to intervene at all levels in obesity control and reduces the relevance of arguments about nature vs. nurture. PMID- 12760447 TI - Gourmand savants and environmental determinants of obesity. AB - Obesity is an embodiment of a multifactorial problem with several intermediates in its casual pathway. Virtually all who have written on obesity have responded to four inter-related factors: genetic, perinatal, environmental, and consumption expenditure energy imbalance. The message to take home is that while a molecular description of each participant of the obesity machinery seems achievable in principle, a complex model describing all of them is currently beyond our grasp. That is why the eradication of the obesity epidemic is seen in a more precise neuropsychological description of what is wrong with each subset of patients. This review proposes that the neuropsychiatric experience might be the most fundamental for it could help to refocus the view of obesity from 'traditional' environmental factors and lifestyle changes to those dominated by a more 'individual-centred' perspective in which different modes of causal attribution are appropriate. This review advocates the idea of environmental dependency as a determinant of obesity, which has been an important idea in neurosciences for more than 30 years with roots in three important areas: psychological, neuropsychiatric, and experimental. The neuropsychology of obesity is yet to become part of today's agenda of obesity research. PMID- 12760448 TI - In vitro mineralization studies with substrate-immobilized bone morphogenetic protein peptides. AB - Understanding the factors that control osteoblastic behavior is centrally important in establishment of successful osseointegration. Pharmacogenetic control of the osteoblast to increase the mineral content around dental implants may offer a unique advantage to clinicians in improving osseointegration success and decreasing time before mechanical loading. This in vitro pilot study has screened for bioactive peptides derived from bone morphogenetic protein 7 (BMP-7) (also called osteogenic protein 1 [OP-1]). Thirteen overlapping peptides of BMP-7 were synthesized and covalently coupled to glass coverslip substrates using silane chemistry. The rate and relative amount of mineralization were compared by von Kossa analysis using primary rat calvarial osteoblastic cell populations during a 7- to 21-day period. In addition, bone sialoprotein (BSP) and osteocalcin (OC) gene expression was measured from osteoblastic cells grown on peptide-immobilized glass coverslips by reverse transcriptase-polymerase chain reaction. Initial results from mineralization studies suggested the BMP-7-derived peptides were able to support mineralization to varying degrees with enhanced peptide-induced mineralization from the C-and N-termini of the BMP-7 molecule. Analysis of these peptide regions indicated that these peptides comprised the finger 1 and 2 domains of CP-1, which contribute toward ligand-receptor interaction. Further analysis of gene expression from select peptide-immobilized substrates indicated that peptides from the C-terminus of BMP-7 were capable of supporting BSP and OC messenger RNA expression. These studies indicate that BMP-7 peptides covalently bound to solid substrates may provide the biological basis to immobilize peptides to titanium implants to induce osteoblastic differentiation and mineralization in a more predictable fashion. PMID- 12760449 TI - Attachment of human marrow stromal cells to titanium surfaces. AB - The attachment of human bone marrow stromal cells to titanium alloy (Ti6Al4V) surfaces was investigated. Titanium disks were polished and modified by surface roughening and by passivation in nitric add. Cell attachment to titanium surfaces and tissue culture plastic (TCP) was determined by tetrazolium bromide (MTT) assay at 2, 6, 24, and 48 hours after seeding. Cell proliferation was determined by thymidine incorporation. Attachment on titanium surfaces was 75.6% to 94.9% of attachment on TCP control. The difference between cell attachment on the TCP compared with smooth or rough titanium was statistically significant (P < .05). However, no statistically significant difference was found between attachment to TCP and passivated titanium. Cell proliferation on titanium surfaces after 24 hours was approximately 70% of proliferation on TCP. There was a statistically significant difference (P < .05) between proliferation on tissue culture and smooth and passivated titanium but not on rough titanium. These results indicate that titanium provides a surface that is conducive to cell attachment and that passivating titanium improves cell attachment, approaching levels seen with TCP, a surface specifically developed to enhance cell attachment. Increasing surface roughness results in improved cell proliferation on titanium. PMID- 12760450 TI - Enhanced osteoblast functions on RGD immobilized surface. AB - Many methods are currently under investigation to improve the integration of dental implants to surrounding bones. Among these methods, peptide-modified surfaces have been highlighted as one of the most promising. Our study, aimed at the cellular response to RGD-immobilized surface in vitro, investigated the basis for designing a bone-active surface coating with RGD-containing peptide. Gold coated titanium surfaces were used as indicative control surfaces for peptide immobilization. Using self-assembly monolayer techniques, 2 types of peptides, RGDC (Arg-Gly-Asp-Cys) and RDGC (Arg-Asp-Gly-Cys), were immobilized onto the gold surfaces. Surface justification was realized through X-ray photoelectron spectroscopy and Fourier transform infrared spectra. Primary calvarial osteoblasts were cultured on RGDC, RDGC, and non-peptide-coated surfaces. Cell attachment, morphology, proliferation, and expression of osteocalcin (OC) messenger RNA (mRNA) were assessed using cell counting, immunolabeling fluorescence microscopy, and Northern blot assay. Four and 8 hours after culture, cell attachment was enhanced on RGDC surfaces. Correspondingly, increased cell spreading and significantly greater cell proliferation were also observed in cells grown on the RGDC-coated surfaces. More importantly, osteoblasts on RGDC surfaces showed earlier and significant OC mRNA expression at day 15 compared with controls having the similar expression at day 21. These results provided evidence of the enhanced functions of osteoblasts cultured on the RGDC-modified surfaces, which might be effective in improving osseointegration for dental implants. PMID- 12760451 TI - Bacterial adhesion on titanium nitride-coated and uncoated implants: an in vivo human study. AB - Titanium nitride (TiN) has been used in many fields as a surgical instrument coating that makes the surgical materials more resistant to wear and corrosion. The aim of the present study was an in vivo evaluation of the bacterial adhesion to TiN-coated (test) and uncoated (control) titanium implants. Six patients aged between 21 and 25 years and in excellent systemic health participated in the study. All of the participants gave their informed consent. The participants were selected on the basis of good periodontal health and no signs of mouth breathing. In each of the 6 participants, a removable acrylic device was adapted to the molar-premolar region of each quadrant of the jaws. One 4 x 13 mm titanium implant was glued to the buccal aspect of each device. The plasma spray covered 11.5 mm of the body of the implant, whereas the neck was machined titanium. Test implants were glued to the right devices and control implants were glued to the left devices. After 24 hours, the implants were removed from each device and processed for scanning electron microscopy for evaluation of the machined portion of the implant covered by bacteria. A total of 24 implants were used in this study, 12 test and 12 control. Surface characterization of the machined portion of the neck of the implant was performed on an additional 10 implants (5 test and 5 control). On test implants the implant surface covered by bacteria was significantly lower compared with that of control implants (P = .0001). The surface roughness was similar in both groups. TiN surfaces showed a significant reduction of the presence of bacteria, and this fact could probably be important in the decrease of the inflammation of the peri-implant soft tissues. PMID- 12760452 TI - Regeneration of the alveolar crest using titanium micromesh with autologous bone and a resorbable membrane. AB - Guided bone regeneration (GBR) has been used for the regeneration of bone in conjunction with the placement of oral implants. The aim of the present study was to clinically and histologically evaluate the use of a titanium micromesh and a resorbable membrane in the GBR technique in patients with alveolar crest defects due to periodontitis, trauma, and extractions. Eighteen patients participated in this study, and 50 implants were inserted. The postoperative healing was uneventful, no dehiscences were observed, and all implants were functioning successfully at 7-year follow-up. At reentry, in all cases, the space under the titanium mesh was completely filled by bone. From a clinical point of view, in all patients, no residual bone defects were observed and a significant increase of the alveolar width or height was found. In all cases, a good esthetic result of the restorative procedures was present. PMID- 12760453 TI - An illustrative study of the role of tomograms for the placement of dental implants. AB - Before dental implants are placed, an evaluation of a presurgical bone site with tomograms will reveal information not available from panoramic or periapical radiographs. This article illustrates the importance of making tomograms before the placement of dental implants to determine the actual height, width, inclination, and undercut of alveolar bone; the shape, cortication, and irregularities of crestal alveolar bone; the density of alveolar bone; the relative location of anatomical landmarks, such as mandibular canal, maxillary sinus, nasal fossa, incisive canal, submandibular gland fossa, etc.; the bucco lingual view of dental pathosis; the bucco-lingual evaluation of sinus graft following sinus-lift surgery; and the evaluation of surgically placed dental implants. PMID- 12760454 TI - Systemic inflammatory response syndrome: a new direction? PMID- 12760455 TI - Effect of tumour and chemoradiotherapy on oesophageal motility. AB - BACKGROUND: The contribution of dysmotility to dysphagia in oesophageal cancer is unclear. AIM: To examine oesophageal motility in patients with oesophageal carcinoma and to assess the effect of chemoradiotherapy on motility. METHODS: Stationary manometry and 24-hour pH-metry were performed in 12 patients with oesophageal carcinoma and one week following completion of chemoradiotherapy using 5-fluorouracil (5-FU), cisplatin and 40Gy radiotherapy. RESULTS: All patients had abnormal motility prior to treatment. Peristalsis was impaired in 11 patients with a mean (SD) of 25% (9) of waves normally propagated. Eight patients had 20% or more simultaneous waves. Following chemoradiotherapy, the percentage of waves normally propagated increased from 25% (9) to 521% (10) (p < 0.03) and normal peristalsis was restored in four patients. The percentage of simultaneous waves decreased from 38% (11) to 21.6% (10) (p = 0.129) while the percentage of dropped or increased waves decreased from 20% (11) to 8.3% (4) (p = 0.264). CONCLUSIONS: Oesophageal motility is disturbed in oesophageal cancer. Dysphagia in oesophageal cancer may be partly explained by oesophageal dysmotility. This is improved by chemotherapy. PMID- 12760456 TI - Bronchopleural fistula after pneumonectomy. AB - BACKGROUND: Bronchopleural fistula (BPF) is an infrequent but life-threatening complication after lung resection. The incidence of BPF reported in the literature varies between 0 and 15%. AIM: To evaluate the incidence of BPF after pneumonectomy for primary lung cancer in a single practice using a single technique. METHODS: Hospital records of patients who underwent pneumonectomy from 1 January 1988 to 1 October 1998 were reviewed retrospectively. The bronchial stump was closed using a uniform hand suture technique. There was a total of 157 patients, including 118 males and 39 females with a mean age of 64 years (range 37-78). Sixty-two patients (39.5%) had a right pneumonectomy. RESULTS: Three patients developed BPF (1.9%). All occurred within seven days of the operation. Two of the three patients died. Ventilation for more than 24 hours was found to be the only significant risk factor for the development of BPF (p < 0.001). CONCLUSIONS: Hand suture closure of bronchial stump after pneumonectomy is a safe, reproducible and inexpensive technique, and a low incidence of BPF can be achieved. PMID- 12760457 TI - Contralateral groin exploration is not justified in infants with a unilateral inguinal hernia. AB - BACKGROUND: Contralateral groin exploration in children with unilateral inguinal hernia is still controversial, particularly in infants. The patency rate of processus vaginalis is highest in infants but there are few data on the subsequent risk of contralateral hernia development in infants. In this retrospective study, we aimed to find out the incidence of contralateral inguinal hernia following unilateral inguinal herniotomy in infants aged less than one year. METHODS: All infants who underwent a unilateral Inguinal herniotomy between January 1990 and December 1998 were studied retrospectively. Infants with bilateral hernia (n = 7) were excluded from the study. RESULTS: One hundred and one infants (93 boys and 8 girls) were studied. Median age at operation was 23 (range 2-52) weeks. The herniotomy was right-sided in 75% of the infants. Follow up ranged from three and a half years to 11 years. A contralateral hernia developed in nine infants (9.0%). One of the initial hernias was incarcerated. Median time from operation to occurrence of contralateral hernia was 18 (range 2 60) months. None of the contralateral hernia was incarcerated. Age, sex, incarceration and side of initial hernia did not influence the development of contralateral hernia. CONCLUSION: The low incidence and benign nature of contralateral hernia development in infants undergoing a unilateral inguinal herniotomy does not justify routine contralateral groin exploration. PMID- 12760458 TI - Endoscopic SIS injection into the lower oesophageal sphincter in dogs. AB - AIM: To determine the usefulness of endoscopically-delivered small intestinal submucosa (SIS) as a scaffold in enhancing the lower oesophageal sphincter (LOS) pressures. METHODS: Six dogs were endoscopically injected--four with the SIS and two with its glycerin carrier. Manometry was performed prior to injection and every four weeks post-op. RESULTS: Adequate and site correct injections were made in four dogs. In one dog, significant augmentation of pressures were obtained at four weeks. None had significant changes in pressure at eight weeks, differences in length at either four or eight weeks or significant differences in the thickness of the examined layers. Four of the six had capillary cushions on pathological examination. The dog injected with the carrier had a loose and disorganise collection, while the others were well organised. CONCLUSION: SIS is a biologically compatible material. Lack of an animal model for gastro oesophageal reflux disease (GORD) makes determining the ability of injections of SIS to combat reflux problematic. PMID- 12760459 TI - Methylprednisolone in acute spinal cord injuries. AB - BACKGROUND: Methylprednisolone is the only neuroprotective therapy advocated in acute non-penetrating spinal cord injury. Trials indicate improved neurological outcome following early administration of a high dose regime. The National Spinal Injuries Unit (NSIU) has promoted this regime by a simple laminated poster sent to all Irish A&E departments. AIM: To assess the use of methylprednisolone in patients with spinal cord injuries. METHODS: A retrospective audit of patient data for all patients admitted with traumatic neurological impairment over a 12 month period. RESULTS: One hundred ninety-six patients were admitted during the study period, 28 (14%) received intravenous methylprednisolone of which six had clear records documenting compliance. One patient received both dexamethasone and methylprednisolone in high doses and three had incorrect bolus dosages administered. Six patients received methylprednisolone infusion longer than the protocol, while five patients were given infusions shorter than recommended. Three patients were admitted to the unit that could have received the steroid regime at the point of transfer. CONCLUSIONS: There was poor documentation of prescription orders and timing of administration. Only six patients had clear documentation allowing confirmation of adherence to the protocol of the National Acute Spinal Cord Injury Study (NASCIS) III trial. PMID- 12760460 TI - Minimally invasive techniques in common surgical procedures: implications for training. AB - BACKGROUND: Laparoscopic techniques are increasingly used in common surgical procedures. Many of these procedures are used to teach basic surgical trainees (BST) and therefore introduction of these techniques may have implications for training. AIMS: To establish whether the introduction of laparoscopic techniques reduced the opportunity of BSTs to perform surgical procedures. METHODS: Patients undergoing hernia repair or appendicectomy in 1991 (when laparoscopy was first introduced) and 1997 (when laparoscopy was readily available) were identified using the Hospital In-Patient Enquiry (HIPE) database. The principal operator and whether the procedure was open or laparoscopic were identified by chart review. RESULTS: The data showed a 50% reduction in the number of appendicectomies performed by BSTs following the introduction of laparoscopic techniques. The number of hernia repairs performed by BSTs has been preserved but the proportion by BSTs fell from 10 to 6%. The proportion of BST-performed procedures carried out laparoscopically has been reduced compared with the registrar-performed group. CONCLUSIONS: The use of minimally invasive techniques has had a negative effect on surgical training. Appropriate measures must be taken to minimise this and such measures should include a structured approach to laparoscopic training and greater access to laparoscopic training facilities. PMID- 12760461 TI - Primary care anticoagulant management using near patient testing. AB - BACKGROUND: Indications for anticoagulant treatment are increasing and new approaches to anticoagulant services require a shift from hospital to primary care. AIM: To pragmatically test the validity and effectiveness of primary care anticoagulant management using near patient testing. METHODS: Twelve CoaguCheck monitors were supplied to 16 rural practices that had previously provided supervision of anticoagulant therapy. Practices were required to record data for eligible patients from September 1998 to April 1999 and to forward one blood sample per week to the regional hospital laboratory for parallel testing. RESULTS: Nine practices returned data on 122 patients. Indications for anticoagulation Included atrial fibrillation (n = 56), valve replacement (n = 12) and deep venous thrombosis or pulmonary embolus (n=12). Regression of the mean of 185 paired readings against their difference confirmed the validity of the CoaguCheck monitor (r2 = 0.00 [95% CI -0.38 to 0.38]). There were 692 International Normalised Ratio (INR) tests performed representing an average of 5.7 tests per patient. The desired therapeutic range was provided for 609 (88%) of these tests; 294 (48.3%) were within the desired therapeutic range. Results differed significantly between practices. CONCLUSIONS: This study confirmed the validity of anticoagulant management using the CoaguCheck monitor in primary care. PMID- 12760462 TI - Rotavirus infection in hospitalised children: incidence and impact on healthcare resources. AB - BACKGROUND AND AIMS: In anticipation of vaccine development this study sought to determine the incidence, disease burden and associated financial burden of rotavirus (RV) infection, in hospitalised children. METHODS: Prospective observational study in two Dublin paediatric centres. RESULTS: Of 663 confirmed infections, 485 (73%) patients were hospitalised with community acquired (CA) RV; 178 (27%) cases were hospital acquired (HA) RV. A total of 243 (50%) children were < 12 months of age, with peak incidence in the 6 to < 12 month age group. CA RV resulted in utilisation of 2,305 bed days, with a median bed stay of three days (range 1-91), representing a minimum cost of Euro176,637 per year to the hospitals. When nosocomial spread and secondary cases are included, this increases to Euro258,695 per year. CONCLUSION: CA RV infection accounted for 1% of all admissions during the study period at a minimum cost of Euro728.40 per case. A safe and effective vaccine could reduce morbidity and advantage children by allowing redeployment of healthcare resources to other critical areas. PMID- 12760464 TI - Malignant phaeochromocytoma metastasising to the breast. AB - BACKGROUND: Nearly one-quarter of metastatic tumours in the breast are from an occult extramammary tumour, usually a lung carcinoma. AIM: To report on a patient with a history of metastatic malignant phaeochromocytoma and a breast mass. RESULT: A 54-year-old female presented with a right breast mass. At the age of 32, she had presented with a phaeochromocytoma. The staining of the breast mass was comparable with that of her original adrenal tumour. CONCLUSION: This is the first published case of a phaeochromocytoma metastasising to the breast, and demonstrates the challenge that extramammary tumours in the breast can pose for the pathologist. PMID- 12760465 TI - Bilateral primary renal lymphoma. AB - BACKGROUND: Primary renal lymphoma is rare with no well-defined diagnostic criteria. AIM: To describe the first reported case of bilateral and primary renal lymphoma and describe diagnostic criteria. RESULT AND CONCLUSION: If a diagnosis of primary renal lymphoma is to be considered, then specific diagnostic criteria (as outlined) must first be satisfied. PMID- 12760466 TI - Opiate overdose in Dublin. PMID- 12760463 TI - Schizophrenia: solving the puzzle. AB - BACKGROUND: Schizophrenia is a common mental illness with an incidence of 15 new cases per 100,000 population per year. AIM: To review evidence for current neurodevelopmental models of the aetiology of schizophrenia. METHODS: We performed a literature search using Medline and PsychINFO. We evaluated the relevance of each article and tracked other relevant articles through references. RESULTS: There is considerable evidence to support neurodevelopmental models of the aetiology of schizophrenia. One or more aetiological events occur between conception and birth that disturb central nervous system (CNS) development, leading to persisting alterations in brain structure and function. These early events, acting in concert with genetic loading and later influences or insults, predispose to the development of schizophrenia in early adulthood. CONCLUSIONS: There have been considerable advances in schizophrenia research over the past 20 years. Future study of Indices of neural development will help advance our understanding of this common, disabling mental illness. PMID- 12760467 TI - Cannabis abuse in Iran. PMID- 12760468 TI - Prostatic carcinoma presenting as testicular mass. PMID- 12760469 TI - Renal cell carcinoma metastasis to breast. PMID- 12760470 TI - Childhood corrosive stomach injury. PMID- 12760471 TI - Dextran sulfate protects porcine but not bovine cultured endothelial cells from free radical injury. AB - Previous studies demonstrated that the polyanion dextran sulfate (DS) protects rat coronary and porcine aortic endothelium (PAE) from oxygen-derived free radical (OFR) injury due to hydrogen peroxide (H2O2) or xanthine/xanthine oxidase (X/XO). To determine if DS has a similar protective effect in bovine aortic endothelium (BAE) and bovine brain microvascular endothelium (BBME), H2O2 or X/XO was added to confluent cultures. Cell injury was assessed 1 d later by measuring the percentage of viable cells (by trypan blue exclusion) and the release of lactate dehydrogenase (LDH) into the medium. After H2O2 doses of 6.0 mM for BAE and BBME and 0.8 mM for PAE, and after X doses of 10 microM and XO doses of 0.3 U/mL for all cell types, approximately 50% of cells were viable. Cultures were pretreated with DS (0.001 to 500 microg/mL) 24 to 26 h prior to H2O2 or X/XO exposure. Pretreatment at concentrations of 0.5, 5, and 50 microg/mL significantly increased the percentage of viable cells and reduced LDH release in cultures of PAE, but not BAE or BBME, treated with H2O2. Similarly, pretreatment with DS concentrations of 5 and 50 microg/mL significantly increased the percentage of viable cells and reduced LDH release in cultures of PAE, but not BAE or BBME, treated with X/XO. Thus, DS protected porcine but not bovine endothelium. Catalase (10 U/mL) increased the percentage of viable cells and reduced LDH release in H2O2-treated BAE and BBME, suggesting that DS likely acts by a different mechanism and does not neutralize H2O2. These results suggest that the protective effect of DS on OFR-injured endothelium is species-dependent. PMID- 12760472 TI - Kinetics and postmucosal effects on urinary recovery of 5 intravenously administered sugars in healthy cats. AB - The objective of this study was to describe the kinetics of urinary recovery and to evaluate the effects of postmucosal factors on urinary recovery of 5 intravenously administered saccharides. Ten cats received an isotonic sugar solution containing lactulose, rhamnose, xylose, methylglucose, and sucrose intravenously. These sugars were selected because of their prior use for intestinal permeability and mucosal function testing in humans and dogs. Urethral catheterization with a closed collection system was used for collection of cumulative urine samples prior to and 2, 4, 6, 8, 10, 12, and 24 h after administration of the sugar solution. High-pressure anion exchange liquid chromatography with pulsed amperometric detection was used to measure the concentrations of each sugar in the urine and calculate urinary recovery. Twenty four hour cumulative urinary recovery for each sugar from the cats, was lower than expected compared to dogs and humans. All 5 sugars had the highest percentage of urinary recovery during the first 2 h after administration. Mean sugar elimination rate constants and half-lives ranged from 0.268/h for methylglucose to 0.415/h for lactulose and 1.67 h for lactulose to 2.59 h for methylglucose, respectively. Metabolism and incomplete urine collection are possible reasons for lower cumulative urinary recoveries of these 5 sugars in cats compared with dogs. Although these 5 sugars are not ideal marker molecules, they may still be useful for intestinal permeability and mucosal function testing in cats. PMID- 12760473 TI - Different central manifestations in response to electroacupuncture at analgesic and nonanalgesic acupoints in rats: a manganese-enhanced functional magnetic resonance imaging study. AB - Acupuncture analgesia is an important issue in veterinary medicine. This study was designed to elucidate central modulation effects in response to electroacupuncture (EA) at different acupoints. Manganese-enhanced functional magnetic resonance imaging was performed in Sprague-Dawley rats after sham acupuncture, sham EA, or true EA at somatic acupoints. The acupoints were divided into 3 groups: group 1, analgesic acupoints commonly used for pain relief, such as Hegu (LI 4); group 2, nonanalgesic acupoints rarely used for analgesic effect such as Neiguan (PC 6); and group 3, acupoints occasionally used for analgesia, such as Zusanli (ST 36). Image acquisition was performed on a 1.5-T superconductive clinical scanner with a circular polarized extremity coil. The results showed that there was no neural activation caused by EA at a true acupoint with shallow needling and no electric current (sham acupuncture). When EA at a true acupoint was applied with true needling but no electric current (sham EA), there was only a slight increase in brain activity at the hypothalamus; when EA was applied at a true acupoint with true needling and an electric current (true EA), the primary response at the hypothalamus was enhanced. Also, there was a tendency for the early activation of pain-modulation areas to be prominent after EA at analgesic acupoints as compared with nonanalgesic acupoints. In conclusion, understanding the linkage between peripheral acupoint stimulation and central neural pathways provides not only an evidence-based approach for veterinary acupuncture but also a useful guide for clinical applications of acupuncture. PMID- 12760474 TI - Serum C-reactive protein in dairy herds. AB - The purpose of this study was to determine the relationship between the serum level of C-reactive protein (CRP) and lactation and health status. Blood samples were collected every 2 wk for 12 mo from 29 randomly selected dairy cattle on 3 farms. At the time the blood samples were collected, the stage of pregnancy, lactation status, breeding records, general health condition, reproductive status, and body condition score were recorded for each cow. Serum CRP was detected with sodium dodecyl sulfate polyacrylamide gel electrophoresis and western immunoblotting. C-reactive protein levels were measured with a densitometer and expressed as an optimal dose value. C-reactive protein levels were correlated with the body condition score, lactation status, and animal health (P < 0.05), but not with ambient temperature, animal age, or parity. C reactive protein levels increased with milk production, peaking during high lactation (2 to 4 mo of pregnancy), and decreased when lactation ceased. In addition, the CRP level was highest during naturally occurring infections, such as mastitis and other tissue inflammation. Thus, the CRP level can confirm the presence of inflammation. The stress effect of taking blood samples as measured by the CRP level, was also examined. The CRP level became rapidly elevated 12 h after the blood samples were taken but returned to normal 36 h later. In conclusion, the stresses resulting from overall poor health, heavy lactation, and blood sampling caused the elevation of serum CRP. C-reactive protein is a marker or tool for evaluating the health status of a herd. C-reactive protein should also be considered as a useful criteria to assess the stress levels and may be useful in early surveillance of disease conditions in a dairy herd. PMID- 12760475 TI - Isolation and characterization of porcine circovirus type-2 from sera of stillborn fetuses. AB - In order to examine an association between porcine circovirus type-2 (PCV2) infection and reproductive failure in pigs, sera (n = 171) from stillborn fetuses were collected from 3 different farms with prolonged histories of reproductive problems. These sera were tested for the presence of antibodies to PCV2 using an immunoperoxidase monolayer assay. Of the 171 sera tested, 28 had PCV2 antibody titers of > or = 1:16. When these 28 samples were tested by a polymerase chain reaction assay,13 were found to contain PCV2 viral DNA. Of these 13 samples containing both PCV2 antibodies and viral DNA, 9 yielded PCV2 on virus isolation. Amino acid sequences comprising open reading frame 2 of PCV2 from 2 of these isolates were compared to PCV2 isolates from cases of post-weaning multi-systemic wasting syndrome (PMWS). The amino acid sequences of the 2 isolates from stillborn pigs were shown to be nearly identical to each other, as well as to other PCV2 isolates associated with reproductive failure. When compared with PMWS isolates, the isolates from the stillborn fetuses showed differences of at least 2 amino acids. These results confirm previous findings that transplacental infection of PCV2 occurs in the field and that stillbirths in pigs may be associated with PCV2 infections. At present, the significance of minor differences in amino acid sequences is not known. PMID- 12760476 TI - Comparative serologic and virologic study of commercial swine herds with and without postweaning multisystemic wasting syndrome. AB - A comparative serologic and virologic study was performed in pigs from 5 herds with postweaning multisystemic wasting syndrome (PMWS) and 2 herds without PMWS in Quebec. In each herd, 60 blood samples were collected at 4-wk intervals from pigs from 3 to 23 wk of age. The serum was evaluated for the presence of antibodies to porcine circovirus type 2 (PCV2) and porcine reproductive and respiratory syndrome virus (PRRSV), as well as for the presence of nucleic acid of PCV2, PRRSV, an porcine parvovirus (PPV), by means of the polymerase chain reaction (PCR). Serologic profiles for PCV2 were very similar in 6 of the 7 herds, including the 2 without PMWS, and were characterized by a gradual decrease in antibody titres from 3 until 11 wk of age, followed by seroconversion at 15 wk, and high PCV2 antibody titres thereafter in all pigs. Only starting at 11 to 15 wk of age could PCV2 viremia be detected, except in 1 herd, in which clinical signs were observed at 6 to 7 wk of age. A PCV2 viremia could be detected within the same pigs for a minimum of 8 wk, and the virus could still be detected in 41% of the serum samples obtained at 23 wk of age. The antibody level did not appear to influence the occurrence of disease, since titres were similar in pigs in the herds with or without PMWS. Infection with PRRSV, as demonstrated by PCR and seroconversion, preceded that of PCV2 by at least 1 mo in both types of herd. Both PRRSV and PCV2 were detected in some pigs in 5 of the 7 herds, including 1 herd without PMWS. Porcine parvovirus could be detected in serum by PCR in 2 herds with PMWS after the onset of clinical signs and also in 1 herd without PMWS. Genomic analysis of PCV2 strains identified in the herds without PMWS indicated complete or very high homology (99.4% to 100%) with the PCV2 strains identified in 4 herds with PMWS. In our field study, the triggering of PMWS in the herds could not be linked to coinfection with either PRRSV or PPV or to the use of a specific immunostimulant, such as vaccines, or to particular genomic differences between the PCV2 strains identified. PMID- 12760477 TI - Strain predominance following exposure of vaccinated and naive pregnant gilts to multiple strains of porcine reproductive and respiratory syndrome virus. AB - Two studies were performed in order to test the relative ability of different strains of porcine reproductive and respiratory syndrome virus (PRRSV) to replicate and cross the placental barrier in pregnant gilts. Study 1 comprised 6 nonvaccinated gilts. Study 2 comprised 8 nonvaccinated gilts and 12 gilts that were vaccinated twice before conception. On, or about, gestation day 90 all gilts were simultaneously exposed to 20 field strains of PRRSV (all strains were distinguishable by restriction fragment length polymorphism (RFLP) patterns). Gilts of study 1 were euthanized on day 7 postpartum. Gilts of study 2 were euthanized on, or about, gestation day 111. All gilts, pigs, and fetuses were tested for the presence and type of strain of PRRSV. Of 128 samples shown to contain PRRSV, 118 contained a single strain, 4 contained 2 strains, and 2 contained a strain or strains for which the RFLP pattern was undecipherable. Only 8 of the 20 strains were isolated from nonvaccinated gilts and their litters. And only 2 of the 20 strains (notably 2 of the same strains isolated from nonvaccinated gilts and their litters), were isolated from vaccinated gilts and their litters. Moreover, 1 of the 2 strains accounted for most (31 of 37; 84%) of the isolates from the vaccinated group. Collectively these results indicate that strains differ in their ability to replicate in pregnant gilts and cross the placental barrier. And they suggest that maternal immunity, although sometimes insufficient to prevent transplacental infection, can exert additional selective pressure. PMID- 12760478 TI - Comparison of assays for the detection of West Nile virus antibodies in chicken serum. AB - Six tests for the detection of West Nile virus (WNV) antibodies in the serum of experimentally infected chickens were compared. The tests included the hemagglutination-inhibition test (HIT), immunoglobulin M (IgM)-capture enzyme linked immunosorbent assay (ELISA) with WNV-infected mouse brain antigen, immunoglobulin G (IgG) indirect ELISA with tickborne encephalitis viral antigen, the microtitre virus neutralization test, the standard plaque reduction neutralization test (PRNT), and the microtitre PRNT (micro-PRNT). Thirty adult chickens, intravenously and intramuscularly inoculated with 10(7) plaque-forming units (PFU) of WNV strain Egypt 101, were bled and given a booster of 10(7) PFU at 7,15, and 21 d postinoculation; the final blood collection was on day 28. Although the micro-PRNT is capable of detecting the highest antibody titres during both early and late infection, because of the technical complexity and time requirements of this test a combination of IgM and IgG ELISAs is recommended for serologic screening. Serum samples that give positive results in the ELISAs can then be tested by the micro-PRNT to determine the specificity of antibodies to WNV. PMID- 12760480 TI - Sample collection factors affect the sensitivity of the diagnostic test for Tritrichomonas foetus in bulls. AB - The current diagnostic test for Tritrichomonas foetus involves the culture of collected preputial or vaginal samples. In an earlier study, which evaluated sampling tools for use with bulls, it was observed that the sensitivity of the diagnostic test was higher for 2nd samples collected from the right side of the prepuce than it was for samples collected 1st from the left side. The study described in this paper was conducted to evaluate which of these factors was responsible for the effect on diagnostic sensitivity. Twenty-nine bulls infected with T. foetus were repeatedly sampled in a 2-factor cross-over design. Samples taken from the right side of the prepuce were 4 times as likely to be positive as samples taken from the left side (P = 0.03). Other factors did not have a significant effect on the outcome of the diagnostic test. Unexpected factors may affect the sensitivity of the diagnostic test for T. foetus. PMID- 12760479 TI - Multiplex nested PCR compared with in situ hybridization for the differentiation of porcine circoviruses and porcine parvovirus from pigs with postweaning multisystemic wasting syndrome. AB - Multiplex nested polymerase chain reactions (PCRs) were developed for the simultaneous detection and differentiation of genomic material of porcine circovirus 1 (PCV1), porcine circovirus 2 (PCV2), and porcine parvovirus (PPV) in formalin-fixed, paraffin-embedded tissues. Multiplex conventional and nested PCR and in situ hybridization were compared for their ability to detect the 3 viruses in such tissues. Xylene deparaffinization followed by proteinase K digestion yielded DNA of sufficient quality for reliable and consistent PCR analyses. The DNA from PCV1, PCV2, and PPV was detected by both multiplex nested PCR and in situ hybridization in lymph-node tissue from 12 pigs experimentally co-infected with the 3 viruses, as well as in formalin-fixed, paraffin-embedded lymph-node tissue from 30 pigs with naturally occurring postweaning multisystemic wasting syndrome; the agreement rates for the 2 methods were 100% in both groups of pigs. Thus, multiplex nested PCR could be applied successfully to formalin-fixed, paraffin-embedded tissues for simultaneous detection of these 3 porcine viruses. PMID- 12760481 TI - Determination and correlation of anti-Neospora caninum antibodies in dogs and cattle from Mexico. AB - The aim of the present study was to determine and to compare through an indirect enzyme linked immunosorbent assay (ELISA) test, the presence of anti-Neospora caninum antibodies in city and farm dogs, as well as in farm cows, and the relationship among them. The correlation between anti-N. caninum antibodies in farm dogs and cattle was also assessed. The research was conducted in the dairy region of Tizayuca, Hidalgo, Mexico. The frequency of anti-N. caninum antibodies was significantly higher in farm dogs (n = 14) (51%) when compared to those from the city (n = 6) (20%) (P < 0.05), suggesting that farm dogs have a higher risk of exposure to the parasite. There was no significant difference in seropositivity between males (n = 11) (39%) and females (n = 9) (33%) (P > 0.05). The frequency of anti-N. caninum antibodies in farm cattle was significantly higher in farms with dogs (n = 158) (58%) when compared to those with no dogs (n = 43) (35%) (P < 0.05). These results suggest the possible transmission of the parasite from dogs to cattle. PMID- 12760482 TI - Naturally-farrowed, artificially-reared pigs as an alternative model for experimental infection by Haemophilus parasuis. AB - The use of naturally-farrowed, artificially-reared piglets as an alternative model to study Haemophilus parasuis infections was evaluated. Two trials were performed in order to evaluate the proposed model. In trial 1, animals were vaccinated and challenged with H. parasuis. Results showed that the proposed model was effectively used to evaluate protective immunity against this organism. In trial 2, animals were challenged with different doses of H. parasuis. Results showed that the severity of clinical signs and lesions tended to increase with higher doses. The reproduction of clinical signs and lesions characteristic of H. parasuis systemic infection was successful in both trials, proving that this model is a viable alternative to specific-pathogen free and cesarean-derived, colostrum-deprived pigs. PMID- 12760483 TI - Effect of oocyte quality on the relative abundance of specific gene transcripts in bovine mature oocytes and 16-cell embryos. AB - Although the developmental potential of oocytes is related to oocyte quality, whether the expression of specific genes is altered in oocytes of different quality and in resulting embryos is not known. Semi-quantitativereverse transcription-polymerase chain reaction was used to compare the relative abundance of 2 transcripts for housekeeping proteins (beta-actin and ribosomal protein L30) and 3 transcripts for growth factor ligand or receptors (platelet derived growth factor receptor alpha (PDGFRalpha), basic fibroblast growth factor (bFGF)), in mature bovine oocytes of high versus low developmental potential. The transcripts for L30, PDGFRalpha, and bFGF in 16-cell embryos originating from these oocytes were also examined. No significant effect of oocyte quality was detected for any of the transcripts examined from oocytes or 16-cell embryos. In conclusion, a lower developmental potential of oocytes with advanced signs of atresia, was not associated with a lower level of abundance of the transcripts examined. PMID- 12760484 TI - Low doses of dexamethasone decrease brain water content of collagenase-induced cerebral hematoma. AB - Different doses of dexamethasone were evaluated for the treatment of cerebral trauma using a rat model of cerebral hematoma induced by intracerebral (IC) stereotaxic injections of collagenase. Control animals received an intracerebral collagenase injection followed by intraperitoneal (IP) saline injection. Sham operated animals received saline only (IC, IP). Forty-eight hours following the surgeries, the brains were removed from the euthanized animals. Cerebral hemispheres were separated and the 4 coronal sections (antero-posterior plane) were weighed. Each slice was dried for 24 h (100 degrees C) and weighed again to establish brain water content. In hematoma-induced saline treated rats, significant differences in brain water content were observed when compared to sham operated animals. Rats treated with 1 mg/kg dexamethasone had a significant brain water content decrease; however, no significant differences were observed with higher doses of dexamethasone. In conclusion, low doses of dexamethasone seem to be beneficial for the treatment of cerebral trauma. PMID- 12760485 TI - Induction of renal metallothionein in rats with ischemic renal failure. AB - Metallothionein (MT) is induced by various types of oxidative stress. However, whether or not MT is induced in renal ischemia/reperfusion injury, in which oxidative stress is believed to play a major role, remains unknown. The present study investigated MT expression in the kidneys of rats with ischmic acute renal failure (IARF). Rats were subjected to 60 min of bilateral renal ischemia followed by reperfusion. Renal MT mRNA expression was then analyzed by Northern blotting. MT expression in ischemic kidney was also localized by in situ hybridization and immunohistochemistry. Renal MT mRNA expression, which was barely detectable in the sham-operated control kidney, increased significantly at 3 h afer reperfsion, continued to increase to a maximal level at 24 h that was maintained for 48 h. The level of MT mRNA expression returned to that of the control by day 4. A morphological study revealed that MT was expressed exclusively in the renal tubular epithelial cells, which are the targets of ischemia/reperfusion injury, and that MT predominated in the outer medulla in the IARF rat kidney at transcriptional and translational levels. These results suggest that MT induced in the IARF rat kidney plays an important role in protecting renal cells against oxidative stress induced by ischemia/reperfusion. PMID- 12760486 TI - Induction of hepatic DT-diaphorase in mice by butylated hydroxyanisole analogs: a structure-activity study. AB - Butylated hydroxyanisole (BHA) and its analogs were evaluated for their relative activity to induce hepatic DT-diaphorase (EC 1.6.99.2) after dietary administration (at concentrations of 11.1 or 27.7 micromol/g diet for 3 days) to mice. Of the compounds tested, only BHA and 2-tert-amyl-4-methoxyphenol, 4 methoxyphenols with 2-tert-alkyl side chains, were active in inducing DT diaphorase activity. None of the remaining six compounds showed any significant sign of inducing activity. No simple explanation for these rather strict structural requirements can be offered at the present time. PMID- 12760487 TI - Effect of lonidamine on alpha2-macroglobulin, hemopexin and alpha1-antitrypsin in the rat testis and epididymis. AB - In a recent study (Leone et al., 2000) we reported that lonidamine (LND), an antispermatogenic drug, affected the concentration of selected testicular and epididymal proteins in the rat. Thus, the effect of LND on alpha2-macroglobulin (alpha2-M) and on other two acute phase proteins (APP), hemopexin (HPX) and alpha1-antitrypsin (alpha1-AT) was examined here. LND was administered orally at the dose of 100 mg/kg, the animals were killed after 24 and 48 hr and the samples were analyzed by immunoblotting. The drug did not induce any significant change of alpha2-M in the serum or testis and of HPX and alpha1-AT in the serum, testis or epididymis. Thus, the antispermatogenic action of LND was not accompanied by a significant change of these inflammatory markers, even if it did cause a decrease of alpha1-inhibitor-3, a negative APP, as previously reported (Leone et al., 2000). PMID- 12760488 TI - Disposition and functions of clarithromycin in human THP-1 monocytes during stimulated and unstimulated conditions. AB - The macrolide antibiotics are bacteriostatic agents interfering with protein synthesis but they are taken up by phagocytic cells, e.g. macrophages, neutrophils and fibroblasts which take up infectious organisms into phagosome lysosomal vaculoes. Recent studies have suggested that these macrolide antibiotics block the spread of infections by mechanisms associated with the inflammation process. Herein is a study with clarithromycin using human THP-1 monocytes, a phagocytic cell which has not been studied to date. Clarithromycin was rapidly taken up by the monocytes (approximately 1%) utilizing both saturable carrier and passive processes at pH 7.4 but was exclusively passive at pH 6.8 and 5.0. The carrier process was energy and temperature dependent and appeared to be linked to certain ion channels. Efflux of the drug was rapid and complete in 1 hr. Intracellular disposition showed 74% in the cell sap and 11% in the nucleus. Upon stimulation with zymogen A or bacteria significant increases of uptake occurred in the isolated lysosome-phagosomes. Examination showed that initially clarithromycin treatment triggered the release of NO, H2O2, IL-1 and TNFalpha from the monocytes, known mediators of inflammation, but also mediators which cause bacterial cell death or apoptosis. The activity of the monocyte marker hydrolytic enzyme NAG was elevated at this time as well as protein kinase C activity. Treatment from 2-4 hr with clarithromycin appeared to reverse this process in that the chemical mediator release was reduced along with the activities of hydrolytic enzymes, e.g. NAG and cathepsin D with no evidence of lipid peroxidation and protective SOD enzyme activity elevation. The latter effects of the antibiotic would be useful in blocking the spread of infection or inflammation from the original site. The normal bacterial static killing effects of clarithromycin was evident at 24 but not 2 hr in both extracellular free bacteria and those bacteria phagocytosed by the THP-1 monocytes. PMID- 12760489 TI - Blockade of p38 mitogen-activated protein kinase pathway inhibits interleukin-6 release and expression in primary neonatal cardiomyocytes. AB - The induction of interleukin-6 (IL-6) using combined proinflammatory agents (LPS/IFN-gamma or TNF-alpha/IFN-gamma) was studied in relation to p38 mitogen activated protein kinase (MAPK) and NF-kappaB transcriptional factor in primary neonatal cardiomyocytes. When added to cultures of cardiomyocytes, the combined agents (LPS/IFN-gamma or TNF-alpha/IFN-gamma) had stimulatory effect on the production of IL-6 and the elevation was significantly reduced by SB203580, a specific p38 MAPK inhibitor. SB203580 inhibited protein production and gene expression of IL-6 in a concentration-dependent manner. In this study, IFN-gamma enhancement of TNF-alpha-induced NF-kappaB binding affinity as well as p38 MAP kinase activation was observed. However, a specific inhibitor of p38 MAPK, SB203580, had no effect on TNF-alpha/IFN-gamma or LPS/IFN-gamma-induced NF-kappaB activation. This study strongly suggests that these pathways about TNF-alpha/IFN gamma or LPS/IFN-gamma-activated IL-6 release can be primarily dissociated in primary neonatal cardiomyocytes. PMID- 12760490 TI - Reduction of viral myocarditis in mice lacking perforin. AB - Cellular immunity plays an important role and contributes to morbidity and mortality in the development of cardiomyopathy secondary to acute myocarditis. Lymphocytotoxicity has been proposed as due to infiltrated cytotoxic T lymphocytes (CTLs). Perforin is thought to be a major factor responsible for the cytolytic properties of the CTLs. To investigate whether mice lacking perforin have enhanced severity of viral myocarditis and acceleration of myocardial apoptosis, perforin knockout (KO) mice were infected with encephalomyocarditis virus. The average cardiac viral titer in perforin KO mice was not significantly different from that in the wild type mice. The heart weight/body weight ratio in perforin KO mice on days 4 and 12 were the same in the wild type mice (n=4 of each). Hearts in perforin KO mice were smaller than that in perforin + mice and grades of myocardial necrosis and lymphocyte infiltration were significantly reduced than in perforin + mice on day 12 after viral inoculation. In the perforin KO mice, the number of apoptotic cells was significantly higher than in wild type on day 4. Perforin blockade induced the early induction of myocardial apoptosis and suppressed the late onset of myocardial injury by cellular autoimmunity. Blockade of perforin gene expression at an early stage may limit viral myocarditis for therapeutic intervention. PMID- 12760491 TI - Effects of volatile oil constituents of Nigella sativa on carbon tetrachloride induced hepatotoxicity in mice: evidence for antioxidant effects of thymoquinone. AB - Effects of the volatile oil constituents of Nigella sativa, namely, thymoquinone (TQ), p-cymene and alpha-pinene, on carbon tetrachloride (CCl4-indued acute liver injury were investigated in mice. A single dose of CCl4 (15 microl/Kg i.p.) induced hepatotoxicity 24 h after administration manifested biochemically as significant elevation of the enzymes activities of serum alanine transaminase (ALT, EC:2.6.1.2), asparate transaminase (AST, EC:2.6.1.1) and lactate dehydrogenase (LDH, EC: 1.1.1.27). The toxicity was further evidenced by a significant decrease of non-protein sulfhydryl(-SH) concentration, and a significant increase of lipid peroxidation measued as malondialdhyde (MDA) in the liver tissues. Administration of different doses of the TQ (4, 8, 12.5, 25 and 50 mg/Kg i.p.) did not alter the chosen biochemical parameters measured, while higher doses of TQ were lethal. The LD50 was 90.3 mg/Kg (77.9-104.7, 95% CL). Pretreatment of mice with different doses of TQ 1 h before CCl4 injection showed that the only dose of TQ that ameliorated hepatotoxicity of CCl4 was 12.5 mg/Kg i.p. as evidenced by the significant reduction of the elevated levels of serum enzymes as well as hepatic MDA content and significant increase of the hepatic nonprotein sulfhydryl(-SH) concentration. Treatment of mice with the other volatile oil constituents, p-cymene or alpha-pinene did not induce any changes in the serum ALT measured. In addition, i.p. administration of these compounds 1 h before CCl4 injection, did not protect mice against CC4-induced hepatotoxicity. The results of the present study indicate that TQ (12.5 mg/Kg, i.p.) may play an important role as antioxidant and may efficiently act as a protective agent against chemically-induced hepatic damage. In contrast, higher doses of TQ were found to induce oxidative stress leading to hepatic injury. PMID- 12760492 TI - Treatment of lead poisoning with an immobilized chelator comparison with conventional therapy. AB - Lead poisoning is a global public health problem. In pregnant women it may result in developmental delays of the fetus, in children it my produce learning disability. Available chelators are nephrotoxic when eliminated as lead-chelator complexes. For safe removal of lead from the body we developed a "Lead Hemopurifier" (L-HP), a device with an immobilized chelator. In vitro, applied to lead solutions, this device reduced the lead concentration. Applied to dogs with lead intoxication, Lead-HP-s removed lead from the blood; this was continuously replaced by lead from the bones until the skeleton was cleared from lead deposit. Treatment of lead poisoning in dogs with Lead-HP-s compared favorably with Versenate treatment of children with lead toxicity. This report demonstrates the in vivo efficiency and safety of this new detoxfication method. Methods to induce lead poisoning in dogs and procedures to identify lead released from skeletal deposits are described. PMID- 12760493 TI - Combined administration of glucose precursors is more efficient than that of glucose itself in recovery from hypoglycemia. AB - The purpose of the present study was to investigate the effect of the combined administration of hepatic gluconeogenic substrates (glycerol + L-lactate + L alanine + L-glutamine) on glucose recovery during insulin induced hypoglycemia (IIH), in rats. IIH was obtained by an ip injection of regular insulin (1 U/kg). Thus, 150 min after insulin administration the rats received an ip injection of glycerol + L-lactate + L-alanine + L-glutamine (each 100 mg/kg). In these experiments control groups, which received saline, glucose or isolated precursors (100 mg/kg), were employed. Glycemia was measured 30 min later, i.e., 180 min after insulin injection. The results showed that the combined administration of gluconeogenic precursors is more efficient than that of glucose itself to promote glycemia recovery. Since, the blood levels of hepatic glucose precursors were decreased (glycerol, L-lactate and L-alanine) or maintained (L-glutamine) during IIH, the ability of the liver to produce glucose from these gluconeogenic substrates was investigated. The results showed that the maximal capacity of the liver to produce glucose from glycerol (2 mM), L-lactate (2 mM), L-alanine (5 mM) and L-glutamine (5 mM) was increased. To L-alanine and L-glutamine, not only the glucose production was increased (P < 0.05) but also the production of L-lactate, pyruvate and urea. Therefore, the results suggest that the decreased availability of glucose precursors, promoted by insulin administration, limits the participation of hepatic gluconeogenesis to glycemia recovery. However, the administration of gluconeogenic precursors could overcome this limitation and promote better glycemia recovery than glucose itself. PMID- 12760494 TI - Multiple immunostaining methods to detect traumatic axonal injury in the rat fluid-percussion brain injury model. AB - Immunohistochemistry using beta-amyloid precursor protein (APP) N-terminus antibodies is routinely used to detect traumatic axonal injury (TAI). The temporal and regional distributions of APP C- and N-terminus immunoreactivity were investigated in rats with experimental brain injury and compared to distribution of neurofilament (NF) immunoreactivity. Male Sprague-Dawley rats underwent right lateral fluid-percussion (FP) brain injury or sham injury. Six FP injury rats and two control rats were transcardially fixed with 10% formalin at 1, 6, 24, and 48 hours, and 1 and 2 weeks after injury and serial 6 microm-thick paraffin sections were prepared. At 6 hours after injury, APP C-terminus immunostaining labeled small neurons and axonal deposits in the injured parasagittal cortex, striatum, thalami, and dorsal medulla, whereas APP N terminus and NF immunostaining showed few axonal deposits in the parasagittal cortex. At 24-48 hours post-injury, marked axonal damage, including axonal swelling and bulbs, was observed in the injured cerebral hemisphere, cerebellar white matter, and medulla. NF immunostaining was most sensitive for axonal damage in the injured deep cortical layers, cerebellum, and medulla. At 1-2 weeks after injury, APP N-terminus immunostaining mainly showed dot-like axonal profiles in the injured thalamus. APP C-terminus immunoreactivity may serve as an early marker of TAI, and the C-terminal fragments of APP may be involved in the evolution of TAI because C-terminal fragments of APP are neurotoxic and pro apoptotic. Multiple immunostaining methods may be required to fully recognize the extent of TAI. PMID- 12760495 TI - In vitro effects of new generation fungal derived nitric oxide donors on rabbit basilar artery. AB - The fungal derived nitric oxide donors, (E)-ethyl-2-[(E)-hydroxyimino]-5-nitro-3 hexenamide (FK409) and N-[(E)-4-ethyl-3-[(Z)-hydroxyimino]-5-nitro-3-hexen-1-yl] 3-pyridinecarboxamide (FR144420), were evaluated for the treatment and prevention of cerebral vasospasm induced by subarachnoid hemorrhage (SAH) by an in vitro study using rabbit basilar artery. The tension-relaxation of a 3 mm-long artery segment was carried out in a micro-tissue organ bath with a real-time recorder to record the tension-relaxation curve. Steady contraction of the specimens was induced by KCI (n = 12) and oxyhemoglobin (oxyHb) (n = 12). Sodium nitroprusside was used for comparison. Each of the agents was added in ascending concentration. Relaxation caused by FK409 and FR144420 was significantly greater (p < 0.05) than that by sodium nitroprusside. Relaxation effects of FK409 and FR144420 on the KCl induced steady contraction were better than those on the oxyHb-induced contraction. FK409 and FR144420 have potential uses for the treatment and prevention of SAH-induced cerebral vasospasm. PMID- 12760496 TI - Differentiation of pituitary adenomas from other sellar and parasellar tumors by 99mTc(V)-DMSA scintigraphy. AB - Pentavalent technetium-99m dimercaptosuccinic acid [99mTc(V)-DMSA] scintigraphy was evaluated for the differentiation of pituitary adenomas, especially non functioning adenomas, from other sellar and parasellar lesions. Diffuse 99mTc(V) DMSA accumulation within the tumor was found in seven of seven non-functioning, three of four growth hormone-secreting, and seven of eight prolactin-secreting adenomas, but only partial accumulation in only two of 16 non-pituitary adenomas and normal pituitary glands. There were no significant relationship between tumor to-background ratios and tumor size or serum hormone level. 99mTc(V)-DMSA scintigraphy showed overall sensitivity of 81% (17/21 cases) for detecting pituitary adenomas, in particular 100% for non-functioning adenomas. 99mTc(V) DMSA may be useful for detecting pituitary adenomas, especially non-functioning adenomas, and for the differentiation of non-functioning pituitary adenomas from other sellar and parasellar lesions. PMID- 12760497 TI - Acute surgery for ruptured dissecting aneurysm of the M3 portion of the middle cerebral artery. AB - A 65-year-old woman presented with a ruptured dissecting aneurysm of the M3 portion of the middle cerebral artery (MCA) manifesting as disturbance of consciousness and motor aphasia. Computed tomography revealed subarachnoid hemorrhage. Emergent angiography demonstrated segmental aneurysmal dilatation of the M3 portion of the left MCA. Infectious aneurysm was excluded. Surgery was performed to prevent repeated hemorrhage from the aneurysm. The lesion was excised and flow to the distal MCA was preserved with an anastomosis of the superficial temporal artery to the MCA. Histological examination confirmed that the aneurysmal dilatation was due to arterial dissection caused by disruption of the internal elastic lamina. Distal dissecting aneurysm may occur in the absence of infectious disease. Such ruptured distal dissecting aneurysm should preferably be treated surgically in the acute stage, immediately after detection of the aneurysm. The parent artery of the proximal and distal sides of the aneurysm should be trapped because of the probable weakness of the arterial wall, and bypass surgery performed to preserve the distal circulation. PMID- 12760498 TI - Repeated thrombus formation immediately following carotid endarterectomy. AB - A 56-year-old male presented with thrombus formation manifesting as cerebral embolic infarction suspected to be caused by hemostasis at the carotid bifurcation, not by the intimal abnormalities or hematological disorders. Thrombus repeatedly and reproducibly appeared at the same area in spite of carotid endarterectomy (CEA). Ultrasonography demonstrated a stenotic lesion of the cervical carotid bifurcation. Medical treatment reduced the stenosis, but the thrombus was repeatedly formed at the same area of the cervical carotid bifurcation. CEA was performed. Histological examination of the specimen found only the underlying thin intima consisting of mild fibrous atheromatic change without ulceration or vascular dissection. Ultrasonography following CEA showed reduced blood flow, indicating hemostasis, and moyamoya appearance in that area. The thrombus had recurred in spite of the medical treatment with anti-platelet agent. This repeated thrombus was gradually dissolved and reduced with anticoagulant therapy. Thrombus causing cerebral embolic stroke and originating at the cervical carotid bifurcation is usually due to the intimal atherosclerotic change such as ulcer formation or vascular dissection. The thrombus in this case was probably formed by hemostasis at the cervical carotid bifurcation and CEA was not effective to prevent recurrence. PMID- 12760499 TI - Acute subdural hematoma after lumboperitoneal shunt placement in patients with normal pressure hydrocephalus. AB - Acute subdural hematoma (SDH) is a rare but disastrous complication after lumboperitoneal shunt placement. Four of 206 adult patients with normal pressure hydrocephalus (1.9%) who underwent lumboperitoneal shunt placement suffered acute SDH following head trauma. The interval between shunt placement and acute SDH was one month to 7 years. Two patients had subdural effusion on computed tomography (CT) at 2- and 6-month follow up. All four patients required assistance in their daily activities before acute SDH onset. The traumatic event was a fall. On admission, CT revealed a large SDH that required surgical removal in two patients, of whom one had manifested subdural effusion after shunt placement. The other two patients had a small SDH. None of the four patients had cerebral contusions. Patients with lumboperitoneal shunts, especially those not capable of independent daily activities, are at risk for acute SDH after even minor head trauma. PMID- 12760500 TI - Reversible bilateral vestibular impairment caused by fourth ventricle epidermoid tumor. AB - A 26-year-old man presented with an epidermoid tumor of the fourth ventricle manifesting as headache with nausea and vertigo. Neurological examination revealed no cerebellar signs, except nystagmus. Bilateral vestibular impairment was identified by the caloric test. The tumor was removed via the midline suboccipital approach. The bilateral peripheral vestibular function recovered remarkably postoperatively. This marked improvement suggests that the bilateral vestibular impairment was caused by compression of the vestibular nuclei by the tumor. PMID- 12760501 TI - Surgical strategy for anterior sacral meningocele. AB - A 25-year-old male presented with an anterior sacral meningocele (ASM) manifesting as repeated urinary tract infections. Surgical correction was completed by simple ligation of the thecal sac next to the ostium via sacral laminectomy, and the thickened filum terminale was sectioned. A 22-year-old female presented with an ASM manifesting as transient difficulty in micturition. Subsequent to sacral laminectomy, the thickened filum terminale was sectioned. However, an aberrant nerve root over the ostium made simple ligation hazardous, so that transdural suture around the ostium was carried out. Complete obliteration was confirmed 5 months after the surgery. Magnetic resonance (MR) imaging could clearly demonstrate the involvement of neurologically important structures. Surgical strategy for ASM based on neurosurgical considerations is proposed, because of the frequent association of caudal spinal cord anomaly as well as presacral mass lesion. Intraoperative assistance systems such as endoscopy for cyst content examination and neurophysiological monitorings are recommended. Several months follow up with MR imaging is required to confirm successful surgical correction. PMID- 12760502 TI - Brucellar spondylodiscitis in the lumbar region. AB - A 59-year-old male farmer presented with a rare case of spondylodiscitis as a manifestation of systemic brucellosis. The patient presented with radicular pain and restricted mobility of the spine due to localized muscle spasm in addition to systemic complaints. Magnetic resonance imaging demonstrated discovertebral involvement at the L4-5 intervertebral space, indicating infectious spondylodiscitis. The Rose-Bengal test was positive and the serum antibody titer was 1/1280. The patient was treated with streptomycin combined with tetracycline plus rifampicin, with complete recovery. Early diagnosis is important and prompt antibrucellar chemotherapy is effective in most cases. Therefore, spondylodiscitis due to brucellosis should be considered in the differential diagnosis of spinal infections. PMID- 12760503 TI - Late bilateral temporal lobe necrosis after conventional radiotherapy. AB - A 63-year-old woman presented with radionecrosis in the bilateral temporal lobes manifesting as dementia about 30 years after undergoing conventional radiotherapy for pituitary adenoma. Computed tomography and magnetic resonance (MR) imaging showed edema and cystic lesions in both temporal lobes. The mass in the left temporal lobe was excised. MR imaging 12 days after surgery showed reduced edema. Her dementia had improved. Radionecrosis usually occurs between several months and a few years after radiotherapy. The incidence of radionecrosis is estimated as 5%, but may be higher with longer follow-up periods. Clinical reports have suggested that larger total doses of radiation are associated with earlier onset of delayed necrosis and the fractional dose is the most significant factor causing cerebral radionecrosis. Radionecrosis can occur long after conventional radiotherapy or stereotactic radiosurgery using a linac-based system or a gamma knife unit. PMID- 12760505 TI - The effect of different types of smoking habits on periodontal attachment. AB - The association between different types of smoking (cigarette, Shesha and Argela) and periodontal attachment loss was investigated in the Riyadh area. The study group included 150 subject 20-60 years of age. Of this, 27.5% were cigarette smokers, 20.3% were Shesha smokers, 20.3% were Argela smokers, and 32% were non smokers (females 24.2%: 74% males). The study was based on clinical examination, periodontal parameters recorded by one examiner (AS) and included measurement of pocket depth (PD 1) followed by measurement of recession from the CEJ to gingival margin (GR 2). Attachment loss (AL) was calculated by adding recession or by subtracting gingival overgrowth to measurements of probing depth. The readings were taken from buccal and lingual surfaces of all teeth except third molars. The influence of age, sex and different types of smoking consumption on these periodontal parameters was statistically evaluated using an analysis of variance (ANOVA). The effect of smoking on periodontitis showed no association with sex, but it was associated with age. By performing both univariate and multivariate tests. P-value for smoking and periodontitis were GR (p = 0.0001), PD (p = 0.0001), and AL (p = 0.0001). The result showed that there is increased loss of attachment in smokers than non-smokers. However, when comparing Shesha smokers to cigarette smokers it was found that Shesha smokers had a greater attachment loss (p = 0.0001), recession (p= 0.0001) and deeper pocket depth (p = 0.001) than cigarette and Argela smokers. These findings suggest that all types of tobacco consumption increase periodontal disease severity and Shesha smoking had a greater effect than cigarette and Argela smoking on disease severity. This study confirms that tobacco is an important risk factor for periodontal disease. Since Shesha smoking is widely used among Saudi subjects of both sexes, additional studies should be conducted to study the effect of Shesha smoking on the periodontium. PMID- 12760504 TI - Antibacterial activity of synthetic human B defensin-2 against periodontal bacteria. AB - The oral epithelium is continuously exposed to a variety of microbial challenges that can cause infectious diseases such as periodontal disease. Human B Defensin 2 (hBD-2) is a cationic antimicrobial peptide with low molecular weight, which is inducible from oral epithelial cells upon either bacterial infection or stimulation with inflammatory cytokines. This peptide has a broad antimicrobial spectrum that includes gram-positive bacteria, gram-negative bacteria, and fungi. Therefore, it is thought that hBD-2 plays an important role as one of natural immunities to bacterial infection. However, its activity is inhibited by body fluids such as serum. The aim of this study was to assess the antibacterial activity of synthetic hBD-2 against oral bacteria in the presence of saliva or serum. The antibacterial activity of synthetic hBD-2 was tested against Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Streptococcus mutans, and Escherichia coli. Antibacterial broth assay and diffusion assay were performed in vitro. The antibacterial activity of hBD-2 was approximately equal to that of minocycline at equimolar concentrations. Furthermore, the activity of hBD-2 remained at 60% in the presence of 80% saliva, whereas no activity remained in the presence of 20% serum. Our results suggest the possibility that synthetic hBD-2 could be useful to prevent infection by periodontal bacteria. PMID- 12760506 TI - Periodontal disease: links with serum lipid/ triglyceride levels? Review and new data. AB - Evidence now supports an intriguing role for serum lipids/triglycerides in modulation of the host immune response to infection. This paper will review the studies that have been conducted in humans and various animal models of the association between elevated serum lipids/triglycerides (i.e. hyperlipidaemia) and periodontitis (PD). We will also present new data in rats that suggests a bi directional relationship between hyperlipidaemia and PD, i.e. that PD itself may be able to modulate serum lipid levels. This review will introduce evidence from other studies that the type of dietary lipids is important in the modulation of the immune/inflammatory response. Overall, the studies presented herein suggest that further investigation using epidemiological and experimental approaches is warranted. PMID- 12760507 TI - The use of tetracycline fibres in the treatment of generalised aggressive periodontitis: clinical and microbiological findings. AB - The purpose of the present study was to evaluate the effects of tetracycline fibres (TCF) as an adjunct to scaling in the treatment of generalised aggressive periodontitis and to compare the effects with mechanical treatment only. Ten patients, 24-39 years old referred for treatment to the Department of Preventive Dentistry, Periodontology and Implant Biology, Aristotle University of Thessaloniki took part in the study. A split-mouth experimental design was used. Measurements referring to bleeding on probing (BOP), pocket depth (PD) and clinical attachment level (CAL) were performed at 12 sites randomly selected. Clinical recordings were made at baseline, 2 and 6 months, after treatment. Subgingival plaque samples were taken for microbiological analysis using the 'checkerboard' DNA-DNA hybridisation technique at baseline, immediately after treatment and at 2 and 6 months. Full-mouth scaling and root planing were performed, with the exception of 2 pre-selected sites, which served as controls. Tetracycline fibres were applied in 5 pockets located in the same half mouth. Analysis of clinical findings showed that mechanical instrumentation in combination with TCF application led to a greater improvement in clinical parameters than scaling and root planing only. Microbial analysis showed a statistically significant greater reduction in the percentages of detection for B. forsythus, P. nigrescens and A. naeslundii genospecies II in pockets where tetracycline fibres were applied. In conclusion, the clinical and microbiological data of the present study suggest that the adjunctive use of TCF improves the clinical response of scaling and root planing in aggressive periodontitis patients. PMID- 12760508 TI - Are infant attachment patterns continuously or categorically distributed? A taxometric analysis of strange situation behavior. AB - Contemporary attachment research is based on the assumption that at least three types of infant attachment patterns exist: secure, avoidant, and resistant. It is not known, however, whether individual differences in attachment organization are more consistent with a continuous or a categorical model. The authors addressed this issue by applying P. E. Meehl's (1973, 1992) taxometric techniques for distinguishing latent types (i.e., classes, natural kinds) from latent continua (i.e., dimensions) to Strange Situation data on 1,139 fifteen-month-old children from the NICHD Study of Early Child Care. The results indicate that variation in attachment patterns is largely continuous, not categorical. The discussion focuses on the implications of dimensional models of individual differences for attachment theory and research. PMID- 12760509 TI - Toward assessing attachment on an emotional security continuum: comment on Fraley and Spieker (2003). AB - The article by R.C. Fraley and S.J. Spieker (2003) serves to remind the discipline of the possible virtues of assessing attachments on continua, a practice that has a long history in attachment research. In this commentary, the author further develops the potential contributions of this approach to assessment and advocates renewed efforts toward assessment of attachments on a single continuum of emotional security. The author contends that theory is essential as a guide for new directions in attachment assessment and that Bowlby's notions of secure base and emotional security provide the needed conceptual foundation for these further developments (E. Waters & E. M. Cummings, 2000). Moreover, challenges that have been made historically to the scoring of attachment on a security continuum are addressed. New means for continuously scoring attachment are advocated as a supplement to the primary direction of categorically assessing attachment patterns. PMID- 12760510 TI - Continuity and change in the measurement of infant attachment: comment on Fraley and Spieker (2003). AB - R.C. Fraley and S.J. Spieker (2003) refocused attention on the important question of continuous versus categorical measurement of infant attachment. This commentary begins with a brief response to Fraley and Spieker's position. Next, it highlights the usefulness of the categorical approach by reviewing some of the major advances in the field that have been fostered by a categorical approach. These include the identification of the disorganized attachment group, the development of the concept of conditional behavioral strategies, the creation of systems for coding attachment beyond infancy, and a greater understanding of cross-generational associations. The author then makes several propositions for future research that are necessary for clarifying how best to measure infant attachment. PMID- 12760511 TI - Attachment categories as reflections of multiple dimensions: comment on Fraley and Spieker (2003). AB - Ainsworth's system of classifying patterns of attachment behavior has served the field well and will not be easily displaced--not because of a commitment by attachment researchers to the taxonomic status of these categories but rather because of difficulties lying in the way of a dimensional approach. Foremost among these is the large number of dimensions used in making classifications and the need to develop reliable scales to tap them. Other problems include the need to capture how behavior changes across age and across the episodes of the Strange Situation. Others will debate R.C. Fraley and S.J. Spieker's (2003) arguments concerning taxonomic status. However, whether taxa or not, Ainsworth's categories at the least have represented well the multidimensional space underlying attachment behavior in conditions of mild stress. Only when an alternative approach is demonstrated to predict (with equal power, in practice) the vast range of outcomes associated with Ainsworth's categories will it be a candidate for substitution. PMID- 12760512 TI - Are there really patterns of attachment? Comment on Fraley and Spieker (2003). AB - Ainsworth's description of attachment patterns in the Strange Situation is one of the best known and most enduring descriptive insights in developmental psychology. Yet attachment theorists have paid little attention to whether ABC classifications represent a true taxonomy or to mechanisms that might produce truly distinct patterns of attachment. This comment focuses on three questions. Does attachment theory require distinct patterns of attachment? How can taxonomic analysis contribute to an understanding of individual differences in attachment security? And are attachment theorists asking the right questions? The authors conclude that attachment theory is indifferent to the structure (taxonomic or dimensional) of individual differences. Nonetheless, taxonomic search methods can make important contributions to attachment study if research is broadened to include secure base behavior in naturalistic settings. PMID- 12760513 TI - It's about timing and change: pubertal transition effects on symptoms of major depression among African American youths. AB - Effects of early physical maturation and accelerated pubertal changes on symptoms of major depression were examined in 639 African American children. Three rival hypotheses, early timing, off-time, and stressful change, were tested using 2 waves of data (mean ages = 11 and 13 years). The pubertal effect operates differently according to children's gender and age. For girls, early maturation was consistently associated with elevated levels of depressive symptoms. For boys, early maturers manifested elevated levels of depression only at age 11, but these symptoms subsided by age 13. Boys who experienced accelerated pubertal growth over time displayed elevated symptom levels. Results support the early timing hypothesis for girls and the stressful change hypothesis for boys. Time at assessment is critical when examining boys' pubertal transition. PMID- 12760514 TI - Prenatal androgens and gender-typed behavior: a study of girls with mild and severe forms of congenital adrenal hyperplasia. AB - Gender-typed behaviors and interests were investigated in 26 girls, aged 2-10 years, affected with congenital adrenal hyperplasia (CAH) and in 26 unaffected girls matched for age. Girls with CAH were more interested in masculine toys and less interested in feminine toys and were more likely to report having male playmates and to wish for masculine careers. Parents of girls with CAH rated their daughters' behaviors as more boylike than did parents of unaffected girls. A relation was found between disease severity and behavior indicating that more severely affected CAH girls were more interested in masculine toys and careers. No parental influence could be demonstrated on play behavior, nor did the comparison of parents' ratings of wished for behavior versus perceived behavior in their daughters indicate an effect of parental expectations. The results are interpreted as supporting a biological contribution to differences in play behavior between girls with and without CAH. PMID- 12760515 TI - Does quality of child care affect child outcomes at age 4(1/2)? AB - Research reveals associations between child-care quality and child outcomes. But are these associations causal? Data from the National Institute of Child Health and Human Development (NICHD) Study of Early Child Care. a longitudinal study of children from birth to age 4(1/2), were used to explore 5 propositions that would support a causal argument. Three propositions received support. principally in the cognitive domain: (a) Associations between quality and outcomes remained even with child and family factors controlled; (b) associations between care and outcomes were domain specific; and (c) outcomes were predicted by quality of earlier care with concurrent care controlled. The 4th proposition, that associations between quality and outcomes would be significant with earlier abilities controlled, received limited support. There was no support for the 5th proposition, that quality and outcomes would exhibit dose-response relations. PMID- 12760516 TI - Interpersonal relatedness, self-definition, and their motivational orientation during adolescence: a theoretical and empirical integration. AB - The authors examined a theoretical model linking interpersonal relatedness and self-definition (S.J. Blatt, 1974), autonomous and controlled regulation (E. L. Deci & R. M. Ryan, 1985), and negative and positive life events in adolescence (N = 860). They hypothesized that motivational orientation would mediate the effects of interpersonal relatedness and self-definition on life events. Self-criticism, a maladaptive form of self-definition, predicted less positive events, whereas efficacy, an adaptive form of self-definition, predicted more positive events. These effects were fully mediated by the absence and presence, respectively, of autonomous motivation. Controlled motivation, predicted by self-criticism and maladaptive neediness, did not predict negative events. Results illustrate the centrality of protective, pleasure-related processes in adaptive adolescent development. PMID- 12760517 TI - Pattern induction by infant language learners. AB - How do infants learn the sound patterns of their native language? By the end of the 1st year, infants have acquired detailed aspects of the phonology and phonotactics of their input language. However, the structure of the learning mechanisms underlying this process is largely unknown. In this study, 9-month-old infants were given the opportunity to induce specific phonological patterns in 3 experiments in which syllable structure, consonant voicing position, and segmental position were manipulated. Infants were then familiarized with fluent speech containing words that either fit or violated these patterns. Subsequent testing revealed that infants rapidly extracted new phonological regularities and that this process was constrained such that some regularities were easier to acquire than others. PMID- 12760518 TI - Children's peer selection: experimental examination of the role of self perceptions. AB - Although previous studies have examined characteristics of children selected as friends, little research has examined the role played by characteristics of the selecting child. In 2 experimental studies that examined the role of self perceptions in peer selection, participants (91 seventh graders in Study 1 and 83 third graders in Study 2) viewed various evaluations of themselves. Participants either believed evaluations were written by unfamiliar peers (Study 1) or were asked to imagine that the views of puppets were views of unfamiliar peers (Study 2). Participants were asked to select the peers they wished to meet and interact with. When evaluations were related to specific competence domains, 7th graders preferred positive peers to negative peers, whereas 3rd graders selected peers who viewed them as they viewed themselves. When evaluations were related to global self-worth, children's selections were unrelated to views of their own global worth. Selection of a globally negative peer was associated with attachment-insecurity/maternal-rejection and depressive symptoms. PMID- 12760519 TI - From children's hands to adults' ears: gesture's role in the learning process. AB - Children can express thoughts in gesture that they do not express in speech--they produce gesture-speech mismatches. Moreover, children who produce mismatches on a given task are particularly ready to learn that task. Gesture, then, is a tool that researchers can use to predict who will profit from instruction. But is gesture also useful to adults who must decide how to instruct a particular child? We asked 8 adults to instruct 38 third- and fourth-grade children individually in a math problem. We found that the adults offered more variable instruction to children who produced mismatches than to children who produced no mismatches- more different types of instructional strategies and more instructions that contained two different strategies, one in speech and the other in gesture. The children thus appeared to be shaping their own learning environments just by moving their hands. Gesture not only reflects a child's understanding but can play a role in eliciting input that could shape that understanding. As such, it may be part of the mechanism of cognitive change. PMID- 12760520 TI - Patterns of knowledge in children's addition. AB - Patterns of conceptual and procedural knowledge of addition were examined in 5- to 8-year-olds (N = 80). Conceptual knowledge was measured by assessing children's responses to problems in which addends were reordered or decomposed and recombined. Problems were presented using abstract symbols, numbers, and physical objects. Children were more successful in noticing that addends had been reordered rather than decomposed and in noticing the decomposition of addends presented with objects rather than with symbols. Distinct profiles of procedural competence were derived from an analysis of children's problem-solving accuracy and strategies. Profiles were associated with patterns of conceptual knowledge and with age, although age and conceptual knowledge were not related. Findings highlight the usefulness of identifying profiles of procedural and conceptual knowledge for understanding the development of children's knowledge of addition. PMID- 12760521 TI - Latent growth curve analyses of accelerating decline in cognitive abilities in late adulthood. AB - Latent growth models were applied to data from the Swedish Adoption/Twin Study of Aging to discover if the rate of change in cognitive performance increased from middle age to later adulthood. The sample included 590 participants aged 44 to 88 years at first measurement. Data were gathered at 2 follow-up occasions at intervals of 3 years. Cognitive ability was assessed through 11 tests that tapped crystallized, fluid, memory, and spatial abilities and perceptual speed. Results indicated stability for measures of crystallized ability, linear age changes for many cognitive abilities, and a significant acceleration in linear decline after age 65 for measures with a large speed component. Gender differences were found only in mean level, not in rate of decline. PMID- 12760522 TI - Like mother, like daughter: similarities in narrative style. AB - Children (2-5-year-olds and 8-13-year-olds) and their parents were independently interviewed about highly salient events: injuries serious enough to necessitate hospital emergency room treatment. Free recall narratives were scored using 14 measures of length, elaborative detail, cohesion, coherence,and provision of context. Mothers' narratives were more cohesive and coherent than fathers', and girls' narratives differed from boys' in parallel ways. Parent and child measures were correlated, and narratives of mother-daughter dyads (for the older daughters) showed striking similarity in all 5 properties, whereas there was no narrative similarity within father-son, mother-son, or father-daughter dyads. This suggests a special status for mother-daughter dyads in terms of how events come to be linguistically represented in narrative. PMID- 12760523 TI - Infant visual recognition memory: independent contributions of speed and attention. AB - Relations between infant visual recognition memory and later cognition have fueled interest in identifying the underlying cognitive components of this important infant ability. The present large-scale study examined three promising factors in this regard--processing speed, short-term memory capacity, and attention. Two of these factors, attention and processing speed (but, surprisingly, not short-term memory capacity), were related to visual recognition memory: Infants who showed better attention (shorter looks and more shifts) and faster processing had better recognition memory. The contributions of attention and processing speed were independent of one another and were similar at all ages studied--5, 7, and 12 months. Taken together, attention and speed accounted for 6%-9% of the variance in visual recognition memory, leaving a considerable, but not unexpected, portion of the variance unexplained. PMID- 12760524 TI - Race and the workforce: occupational status, aspirations, and stereotyping among African American children. AB - This study examined whether African American children's perceptions of occupational status and their own vocational interests are affected by racial segregation of the workforce. Children (N = 92) rated familiar occupations with respect to status, desirability, and stereotyping. Children also rated novel jobs that had been depicted with African Americans, European Americans, or both African and European Americans. As predicted, for familiar jobs, children's judgments were linked to their knowledge of racial segregation of these jobs. In addition, novel occupations that had been depicted with African Americans were judged as lower in status than the identical occupations that had been depicted with European Americans, demonstrating a causal influence of workers' race on children's judgments. Children's age and socioeconomic background moderated their occupational judgments. PMID- 12760525 TI - Do children's attention processes mediate the link between family predictors and school readiness? AB - The role of attention processes as possible mediators between family environment and school readiness was analyzed with data from 1,002 children and their families. Data on children's sustained attention, impulsivity, and school readiness (i.e., cognitive, achievement, language, and social development) were obtained at 54 months of age, and quality of the family environment was assessed throughout the first 54 months. Mediation tests showed that children's sustained attention partially accounted for the link between family environment and achievement and language outcomes. Impulsivity partially accounted for the link between family environment and achievement, social competence, and externalizing behaviors. The roles of sustained attention and of inhibition of impulsive responding in the relation between family characteristics and school readiness are discussed. PMID- 12760526 TI - Infants use handrails as tools in a locomotor task. AB - In 2 experiments the authors demonstrated that adaptive locomotion can involve means-ends problem solving. Sixteen-month-old toddlers crossed bridges of varying widths in the presence or absence of a handrail. Babies attempted wider bridges more often than narrow ones, and attempts on narrow bridges depended on handrail presence. Toddlers had longer latencies, examined the bridge and handrail more closely, and modified their gait when bridges were narrow and/or the handrail was unavailable. Infants who explored the bridge and handrail before stepping onto the bridge and devised alternative bridge-crossing strategies were more likely to cross successfully. Results challenge traditional conceptualizations of tools: Babies used the handrail as a means for augmenting balance and for carrying out an otherwise impossible goal-directed task. PMID- 12760527 TI - Longitudinal analysis of flexibility and reorganization in early adolescence: a dynamic systems study of family interactions. AB - A dynamic systems (DS) approach was used to study changes in the structure of family interactions during the early adolescent transition period. Longitudinal observational data were collected in 5 waves prior to, during, and after the transition. Boys (n = 149 families) were videotaped problem solving with their parents at 9-10 years old and every 2 years thereafter until they were 18 years old State space grids (a new DS method) were constructed for all families across all waves. Two variables indexing the variability of the family interactions were derived from the grids. As hypothesized, the DS variables revealed a significant quadratic effect related to a peak in variability at 13-14 years of age. PMID- 12760529 TI - Singlet oxygen: the relevance of extracellular production mechanisms to oxidative stress in vivo. AB - Since the physiological relevance of 1O2 independent of photosensitization has been controversial, we review proposed reaction mechanisms for its extracellular production in vivo and discuss the relevance of this production to oxidative stress. We conclude that extracellular 1O2 production by the spontaneous dismutation of O2*- does have physiological relevance. Also, extracellular 1O2 production by the eosinophil peroxidase-H2O2-bromide system could have physiological relevance. As regards the other reactions discussed in this review, the evidence is not sufficient to warrant any conclusions as to the physiological relevance of these to extracellular 1O2 production. What is evident is that the microenvironment will have a significant influence on the success or failure of extracellular 1O2 production. To date, most demonstrations of 1O2 production by physiologically relevant mechanisms have used conditions that minimize competitive reactions. More research demonstrating how physiologically relevant competitive reactions influence extracellular 1O2 production is needed. PMID- 12760528 TI - Longitudinal direct and indirect pathways linking older sibling competence to the development of younger sibling competence. AB - A 4-wave longitudinal model tested direct and indirect links between older sibling (OS; M = 11.7 years) and younger sibling (YS; M = 9.2 years) competence in 152 rural African American families. Data were collected at 1-year intervals. At each wave, different teachers assessed OS competence, YS competence, and YS self-regulation. Mothers reported their own psychological functioning; mothers and YSs reported parenting practices toward the YS. OS competence was stable across time and was linked with positive changes in mothers' psychological functioning from Wave 1 to Wave 2. Mothers' Wave 2 psychological functioning was associated with involved-supportive parenting of the YS at Wave 3. OS Wave 2 competence and Wave 3 parenting were indirectly linked with Wave 4 YS competence, through Wave 3 YS self-regulation. Structural equation modeling controlled for Wave 1 YS competence; thus, the model accounted for change in YS competence across 3 years. PMID- 12760530 TI - Photophysical properties of 5-methylcytidine. AB - Cytosine methylation, which determines the hot spots for DNA photo-damage, is shown to induce a red-shift of the nucleoside absorption spectrum, making the chromophore more vulnerable to solar radiation, and a tenfold increase of the fluorescence lifetime, making excited statereactions more probable. A femtosecond investigation of the excited state deactivation reveals a quite complex mechanism. PMID- 12760531 TI - Human UVA exposures estimated from ambient UVA measurements. AB - The methods presented in this paper allow for the estimation of human UVA exposure using measured UVA irradiance values. Using measured broadband UVA irradiances over the period of a year, it was estimated that for humans in an upright posture and not moving the head with respect to the body, the nose received 26.5% of the available ambient UVA radiation, whilst the shoulders and vertex of the head received 81% and 100% respectively of the available ambient UVA radiation. Measurement of the exposure ratios for a series of solar zenith angles between 90 degrees and 0 degrees will allow extension of this technique to other latitudes. PMID- 12760532 TI - Comparison of human facial UV exposure at high and low latitudes and the potential impact on dermal vitamin D production. AB - The results presented in this paper allow for the estimation of the monthly UV exposure of the human facial region at various locations across the earth. The technique allows a graphical representation of the UV exposures over the face. The erythemal UV exposures as well as the vitamin D exposures to the human facial region have been investigated. The results gained in this paper, for a clear sky and constant ozone indicate that the sun's capability to promote the development of vitamin D in the human body does not follow the erythemal UV irradiances, in particular at high latitudes. For Amsterdam (52 degrees N) in late winter, approximately 20% more UV is required to produce 215 J m(-2) of vitamin D weighted UV than erythemal UV. PMID- 12760534 TI - Photochemical and photophysical behaviour of vitamin E: interaction of its long lived transient photoproducts with carotenoids. AB - Multi-channel, flash kinetic spectroscopy with microsecond time resolution has been used for investigating the interactions between carotenoids and the following photoproducts of alpha-tocopherol (EH) in hexane, methanol, acetonitrile, and dimethyl sulfoxide: (a) the lowest triplet, (b) the tocopherol radical cation, which could be seen only in the polar aprotic solvents acetonitrile and dimethyl sulfoxide, and (c) the neutral tocopheroxyl radical. The first two species reconvert to EH by transferring triplet excitation and positive charge (respectively) to the carotenoid; the third is unreactive. The relevance of these observations to photoprotection and the photoionisation of sterically hindered phenols is pointed out. PMID- 12760539 TI - Photochemically enhanced gene delivery with cationic lipid formulations. AB - Entrapment and degradation of transfecting DNA in endocytic vesicles often hampers the use of lipidic vectors for gene delivery purposes. Photochemical internalisation (PCI) is a technology for achieving light-induced release of DNA trapped inside these vesicles, and therefore represents a way of overcoming the endocytic membrane barrier and improving gene transfer. The technology is based on utilising photosensitizers which localise in the membranes of endocytic vesicles, causing photochemical damages that rupture the vesicles upon illumination. The purpose of this work was to study the effect of PCI on transfection mediated by the cationic lipid N-(2-aminoethyl)-N,N-dimethyl-2,3 bis(tetradecyloxy)-1-propanaminium bromide (betaAE-DMRIE), with or without the helper lipid 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE). It was shown that PCI has no effect on betaAE-DMRIE mediated transfection, whereas it significantly enhances transfection mediated by the combination of betaAE-DMRIE and DOPE. The effect of PCI was highly dependent on the timing of illumination relative to the time of DNA delivery, both regarding the sequence of, and the time between, these two treatments. PMID- 12760542 TI - Is parietin a UV-B or a blue-light screening pigment in the lichen Xanthoria parietina? AB - Xanthoria parietina is a widespread lichen coloured by the orange cortical pigment parietin (= physcion). We studied the pigment content in 60 thalli sampled in 4 habitats along a sun-shade gradient from evergreen boreal forests through open deciduous stands to sea cliffs. The significant positive regression between contents of parietin per unit area and site factors (reflecting the openness of the canopy relative to an open sky) across sampled habitats suggested a photoprotective role of parietin at UV-B and/or blue wavelengths, the two absorbance maxima of parietin. UV-B susceptibility of X. parietina, measured as permanent reductions in photosystem II, decreased highly significantly with increasing parietin content per thallus area. However, as much as three-fold greater UV-B irradiances than ambient daily summer maxima, maintained continuosly for 240 h were required to cause UV-B damage even in thalli of shaded habitats. Since a previous study has documented a high PAR susceptibility of parietin deficient X. parietina in the absence of UV-B, there are reasons to believe that the blue light screening of parietin is functionally more important than the UV-B screening. A strong positive relationship between parietin content per unit area and reflectance at 500 nm allows the parietin content in X. parietina thalli to be assessed non-destructively by reflectance measurements. PMID- 12760543 TI - Formation of the spore photoproduct and other dimeric lesions between adjacent pyrimidines in UVC-irradiated dry DNA. AB - Far-UV irradiation of DNA leads to the formation of several types of dimeric lesions between adjacent pyrimidine bases including cyclobutane dimers, (6-4) photoproducts and Dewar valence isomers In the dry state, an additional specific thymine lesion, the spore photoproduct, is produced. We designed an HPLC-tandem mass spectrometry assay for the detection of the latter lesion. This technique that does not require radio-labelling of DNA allowed the simultaneous quantification of the spore photoproduct and other pyrimidine dimeric photoproducts. Using this approach, the complete distribution of bipyrimidine lesions within UVC-irradiated dry DNA was determined. PMID- 12760544 TI - Detergent effect on cytochrome b559 electron paramagnetic resonance signals in the photosystem II reaction centre. AB - The detergent effect on Cytochrome b559 from spinach photosystem II was studied by electron paramagnetic resonance (EPR) spectroscopy in D1-D2-Cyt b559 complex preparations. Various n-dodecyl-beta-D-maltoside concentrations from 0 to 0.2% (w/v) were used to stabilise the D1-D2-Cyt b559 complexes. Low spin heme EPR spectra were obtained but the g(z) feature positions changed depending on the detergent conditions Redox potentiometric titrations showed a unique redox potential cytochrome b559 form (E'm = + 123-150 mV) in all the D1-D2-Cyt b559 complex preparations indicating that detergent does not affect this property of the protein in those conditions. A similar effect on Cytochrome b559 EPR spectrum was observed in more intact photosystem II preparations independently of their aggregation state. This finding indicates that changes due to detergent could be a common phenomenon in photosystem II complexes. Results are discussed in terms of the environment each detergent provides to the protein. PMID- 12760545 TI - A study of the relationship between the chemical structures and the fluorescence quantum yields of coumarins, quinoxalinones and benzoxazinones for the development of sensitive fluorescent derivatization reagents. AB - To develop new fluorescent derivatization reagents, we investigated the relationship between the chemical structures and the fluorescence quantum yields (phi(f)) of coumarins, quinoxalinones and benzoxadinones. Forty-six compounds were synthesized and their fluorescence spectra were measured in n-hexane, ethyl acetate, methanol and water. The energy levels of these compounds were calculated by combination of the semi-empirical AM1 and INDO/S (CI = all) methods. The deltaE(Tn(n,pi*), S1(pi,pi*)) (the energy gap between the Tn(n,pi*) and S1(pi,pi*) states) values were well correlated with the phi(f) values, which enables us to predict the phi(f) values from their chemical structures. Based on this relationship, 3-phenyl-7-N-piperazinoquinoxalin-2(1H)-one (PQ-Pz) and 7-(3 (S)-aminopyrrolidin-1-yl)-3-phenylquinoxalin-2-(1H)-one (PQ-APy) were developed as fluorescent derivatization reagents for carboxylic acids. The derivatives of the carboxylic acids with PQ-Pz and PQ-APy showed large phi(f) values even in polar solvents, suggesting that these reagents are suitable for the microanalysis of biologically important carboxylic acids by reversed phase HPLC. PMID- 12760547 TI - A linear wavelet filter for parametric imaging with dynamic PET. AB - This paper describes a new filter for parametric images obtained from dynamic positron emission tomography (PET) studies. The filter is based on the wavelet transform following the heuristics of a previously published method that are here developed into a rigorous theoretical framework. It is shown that the space-time problem of modeling a dynamic PET sequence reduces to the classical one of estimation of a normal multivariate vector of independent wavelet coefficients that, under least-squares risk, can be solved by straightforward application of well established theory. From the study of the distribution of wavelet coefficients of PET images, it is inferred that a James-Stein linear estimator is more suitable for the problem than traditional nonlinear procedures that are incorporated in standard wavelet filters. This is confirmed by the superior performance of the James-Stein filter in simulation studies compared to a state of-the-art nonlinear wavelet filter and a nonstationary filter selected from literature. Finally, the formal framework is interpreted for the practitioner's point of view and advantages and limitations of the method are discussed. PMID- 12760548 TI - Multiresolution fMRI activation detection using translation invariant wavelet transform and statistical analysis based on resampling. AB - A new method is proposed for activation detection in event-related functional magnetic resonance imaging (fMRI). The method is based on the analysis of selected resolution levels (a subspace) in translation invariant wavelet transform (TIWT) domain. Using a priori knowledge about the activation signal and trends, we analyze their power in different resolution levels in TIWT domain and select an optimal set of resolution levels. A randomization-based statistical test is then applied in the wavelet domain for activation detection. This approach suppresses the effects of trends and enhances the detection sensitivity. In addition, since TIWT is insensitive to signal translations, the power analysis is robust with respect to signal shifts. The randomization test alleviates the need for assumptions about fMRI noise. The method has been applied to simulated and experimental fMRI datasets. Comparisons have been made between the results of the proposed method, a similar method in the time domain and the cross correlation method. The proposed method has shown superior sensitivity compared to the other methods. PMID- 12760549 TI - Wavelet-based estimation of a semiparametric generalized linear model of fMRI time-series. AB - This paper addresses the problem of detecting significant changes in fMRI time series that are correlated to a stimulus time course. This paper provides a new approach to estimate the parameters of a semiparametric generalized linear model of fMRI time series. The fMRI signal is described as the sum of two effects: a smooth trend and the response to the stimulus. The trend belongs to a subspace spanned by large scale wavelets. The wavelet transform provides an approximation to the Karhunen-Loeve transform for the long memory noise and we have developed a scale space regression that permits to carry out the regression in the wavelet domain while omitting the scales that are contaminated by the trend. In order to demonstrate that our approach outperforms the state-of-the art detrending technique, we evaluated our method against a smoothing spline approach. Experiments with simulated data and experimental fMRI data, demonstrate that our approach can infer and remove drifts that cannot be adequately represented with splines. PMID- 12760550 TI - A versatile wavelet domain noise filtration technique for medical imaging. AB - In this paper, we propose a robust wavelet domain method for noise filtering in medical images. The proposed method adapts itself to various types of image noise as well as to the preference of the medical expert; a single parameter can be used to balance the preservation of (expert-dependent) relevant details against the degree of noise reduction. The algorithm exploits generally valid knowledge about the correlation of significant image features across the resolution scales to perform a preliminary coefficient classification. This preliminary coefficient classification is used to empirically estimate the statistical distributions of the coefficients that represent useful image features on the one hand and mainly noise on the other. The adaptation to the spatial context in the image is achieved by using a wavelet domain indicator of the local spatial activity. The proposed method is of low complexity, both in its implementation and execution time. The results demonstrate its usefulness for noise suppression in medical ultrasound and magnetic resonance imaging. In these applications, the proposed method clearly outperforms single-resolution spatially adaptive algorithms, in terms of quantitative performance measures as well as in terms of visual quality of the images. PMID- 12760551 TI - Platelets: a multiscale approach for recovering edges and surfaces in photon limited medical imaging. AB - The nonparametric multiscale platelet algorithms presented in this paper, unlike traditional wavelet-based methods, are both well suited to photon-limited medical imaging applications involving Poisson data and capable of better approximating edge contours. This paper introduces platelets, localized functions at various scales, locations, and orientations that produce piece-wise linear image approximations, and a new multiscale image decomposition based on these functions. Platelets are well suited for approximating images consisting of smooth regions separated by smooth boundaries. For smoothness measured in certain Holder classes, it is shown that the error of m-term platelet approximations can decay significantly faster than that of m-term approximations in terms of sinusoids, wavelets, or wedgelets. This suggests that platelets may outperform existing techniques for image denoising and reconstruction. Fast, platelet-based, maximum penalized likelihood methods for photon-limited image denoising, deblurring and tomographic reconstruction problems are developed. Because platelet decompositions of Poisson distributed images are tractable and computationally efficient, existing image reconstruction methods based on expectation-maximization type algorithms can be easily enhanced with platelet techniques. Experimental results suggest that platelet-based methods can outperform standard reconstruction methods currently in use in confocal microscopy, image restoration, and emission tomography. PMID- 12760552 TI - Regularization in tomographic reconstruction using thresholding estimators. AB - In tomographic medical devices such as single photon emission computed tomography or positron emission tomography cameras, image reconstruction is an unstable inverse problem, due to the presence of additive noise. A new family of regularization methods for reconstruction, based on a thresholding procedure in wavelet and wavelet packet (WP) decompositions, is studied. This approach is based on the fact that the decompositions provide a near-diagonalization of the inverse Radon transform and of prior information in medical images. A WP decomposition is adaptively chosen for the specific image to be restored. Corresponding algorithms have been developed for both two-dimensional and full three-dimensional reconstruction. These procedures are fast, noniterative, and flexible. Numerical results suggest that they outperform filtered back-projection and iterative procedures such as ordered-subset-expectation-maximization. PMID- 12760553 TI - Nonseparable wavelet-based cone-beam reconstruction in 3-D rotational angiography. AB - In this paper, we propose a new wavelet-based reconstruction method suited to three-dimensional (3-D) cone-beam (CB) tomography. It is derived from the Feldkamp algorithm and is valid for the same geometrical conditions. The demonstration is done in the framework of nonseparable wavelets and requires ideally radial wavelets. The proposed inversion formula yields to a filtered backprojection algorithm but the filtering step is implemented using quincunx wavelet filters. The proposed algorithm reconstructs slice by slice both the wavelet and approximation coefficients of the 3-D image directly from the CB projection data. The validity of this multiresolution approach is demonstrated on simulations from both mathematical phantoms and 3-D rotational angiography clinical data. The same quality is achieved compared with the standard Feldkamp algorithm, but in addition, the multiresolution decomposition allows to apply directly image processing techniques in the wavelet domain during the inversion process. As an example, a fast low-resolution reconstruction of the 3-D arterial vessels with the progressive addition of details in a region of interest is demonstrated. Other promising applications are the improvement of image quality by denoising techniques and also the reduction of computing time using the space localization of wavelets. PMID- 12760554 TI - Robust estimation of ultrasound pulses using outlier-resistant de-noising. AB - A different approach to the problem of estimation of the ultrasound pulse spectrum, which usually arises as a part of ultrasound image restoration algorithms, is presented. It is shown that this estimation problem can be reformulated in terms of a de-noising problem. In this formulation, the log spectrum of a radio-frequency line (RF-line) is viewed as a noisy measurement of the signal that needs to be estimated, i.e., the ultrasound pulse log-spectrum. The log-spectrum of the tissue reflectivity function (i.e., tissue response) is considered as the noise to be rejected. The contribution of the paper is twofold. First, it provides statistical description of the reflectivity function log spectrum for the case, when the samples of the reflectivity function are independent identically distributed (i.i.d.) Gaussian random variables. Moreover, it is shown that the problem of the pulse spectrum recovery is essentially a de noising problem. Consequently, it is suggested to solve the problem within the framework of the de-noising by wavelet shrinkage. Second, a computationally efficient algorithm is proposed for the pulse-spectrum estimation, which can be viewed as a modified version of the classical Donoho's three-step de-noising procedure. This modification is necessary, because of specific properties of the noise to be rejected. It is shown, that whenever the samples of the reflectivity function can be assumed to be i.i.d. Gaussian random variables, the samples of its log-spectrum obey the Fisher-Tippet distribution. For this type of noise, straightforward implementation of the standard de-noising can cause serious estimation errors. In order to overcome this difficulty, an outlier-resistant de noising is performed. The unique properties of this modified de-noising algorithm allow estimating the pulse spectrum adaptively to its properties, as they are continuously influenced by the frequency-dependent attenuation process. The performance of the proposed algorithm is examined in a series of computer simulations. It is shown that this algorithm, developed on the assumption of the "Gaussian" reflectivity function, remains applicable for broader classes of distributions. The results obtained in a series of in vivo experiments reveal superior performance of the novel approach over some of alternative estimation techniques, e.g., cepstrum-based estimation. PMID- 12760555 TI - Ultrasonic liver tissues classification by fractal feature vector based on M-band wavelet transform. AB - This paper describes the feasibility of selecting fractal feature vector based on M-band wavelet transform to classify ultrasonic liver images-normal liver, cirrhosis, and hepatoma. The proposed feature extraction algorithm is based on the spatial-frequency decomposition and fractal geometry. Various classification algorithms based on respective texture measurements and filter banks are presented and tested. Classifications for the three sets of ultrasonic liver images reveal that the fractal feature vector based on M-band wavelet transform is trustworthy. A hierarchical classifier, which is based on the proposed feature extraction algorithm is at least 96.7% accurate in the distinction between normal and abnormal liver images and is at least 93.6% accurate in the distinction between cirrhosis and hepatoma liver images. Additionally, the criterion for feature selection is specified and employed for performance comparisons herein. PMID- 12760556 TI - Highly regular wavelets for the detection of clustered microcalcifications in mammograms. AB - In this paper, we consider the problem of detecting clustered microcalcification in digitized mammograms using new wavelets with a high Sobolev regularity index. We experimentally assess the superiority of the new wavelets when compared with the classical ones. PMID- 12760557 TI - Integrated wavelets for enhancement of microcalcifications in digital mammography. AB - This paper presents a new algorithm for enhancement of microcalcifications in mammograms. The main novelty is the application of techniques we have developed for construction of filterbanks derived from the continuous wavelet transform. These discrete wavelet decompositions, called integrated wavelets, are optimally designed for enhancement of multiscale structures in images. Furthermore, we use a model based approach to refine existing methods for general enhancement of mammograms resulting in a more specific enhancement of microcalcifications. We present results of our method and compare them with known algorithms. Finally, we want to indicate how these techniques can also be applied to the detection of microcalcifications. Our algorithm was positively evaluated in a clinical study. It has been implemented in a mammography workstation designed for soft-copy reading of digital mammograms developed by IMAGETOOL, Germany. PMID- 12760558 TI - Hierarchical active shape models, using the wavelet transform. AB - Active shape models (ASMs) are often limited by the inability of relatively few eigenvectors to capture the full range of biological shape variability. This paper presents a method that overcomes this limitation, by using a hierarchical formulation of active shape models, using the wavelet transform. The statistical properties of the wavelet transform of a deformable contour are analyzed via principal component analysis, and used as priors in the contour's deformation. Some of these priors reflect relatively global shape characteristics of the object boundaries, whereas, some of them capture local and high-frequency shape characteristics and, thus, serve as local smoothness constraints. This formulation achieves two objectives. First, it is robust when only a limited number of training samples is available. Second, by using local statistics as smoothness constraints, it eliminates the need for adopting ad hoc physical models, such as elasticity or other smoothness models, which do not necessarily reflect true biological variability. Examples on magnetic resonance images of the corpus callosum and hand contours demonstrate that good and fully automated segmentations can be achieved, even with as few as five training samples. PMID- 12760559 TI - Three-dimensional encoding/two-dimensional decoding of medical data. AB - We propose a fully three-dimensional (3-D) wavelet-based coding system featuring 3-D encoding/two-dimensional (2-D) decoding functionalities. A fully 3-D transform is combined with context adaptive arithmetic coding; 2-D decoding is enabled by encoding every 2-D subband image independently. The system allows a finely graded up to lossless quality scalability on any 2-D image of the dataset. Fast access to 2-D images is obtained by decoding only the corresponding information thus avoiding the reconstruction of the entire volume. The performance has been evaluated on a set of volumetric data and compared to that provided by other 3-D as well as 2-D coding systems. Results show a substantial improvement in coding efficiency (up to 33%) on volumes featuring good correlation properties along the z axis. Even though we did not address the complexity issue, we expect a decoding time of the order of one second/image after optimization. In summary, the proposed 3-D/2-D multidimensional layered zero coding system provides the improvement in compression efficiency attainable with 3-D systems without sacrificing the effectiveness in accessing the single images characteristic of 2-D ones. PMID- 12760560 TI - Wavelet coding of volumetric medical datasets. AB - Several techniques based on the three-dimensional (3-D) discrete cosine transform (DCT) have been proposed for volumetric data coding. These techniques fail to provide lossless coding coupled with quality and resolution scalability, which is a significant drawback for medical applications. This paper gives an overview of several state-of-the-art 3-D wavelet coders that do meet these requirements and proposes new compression methods exploiting the quadtree and block-based coding concepts, layered zero-coding principles, and context-based arithmetic coding. Additionally, a new 3-D DCT-based coding scheme is designed and used for benchmarking. The proposed wavelet-based coding algorithms produce embedded data streams that can be decoded up to the lossless level and support the desired set of functionality constraints. Moreover, objective and subjective quality evaluation on various medical volumetric datasets shows that the proposed algorithms provide competitive lossy and lossless compression results when compared with the state-of-the-art. PMID- 12760561 TI - Lossy-to-lossless compression of medical volumetric data using three-dimensional integer wavelet transforms. AB - We study lossy-to-lossless compression of medical volumetric data using three dimensional (3-D) integer wavelet transforms. To achieve good lossy coding performance, it is important to have transforms that are unitary. In addition to the lifting approach, we first introduce a general 3-D integer wavelet packet transform structure that allows implicit bit shifting of wavelet coefficients to approximate a 3-D unitary transformation. We then focus on context modeling for efficient arithmetic coding of wavelet coefficients. Two state-of-the-art 3-D wavelet video coding techniques, namely, 3-D set partitioning in hierarchical trees (Kim et al., 2000) and 3-D embedded subband coding with optimal truncation (Xu et al., 2001), are modified and applied to compression of medical volumetric data, achieving the best performance published so far in the literature-both in terms of lossy and lossless compression. PMID- 12760562 TI - A new approach to accelerated drug-excipient compatibility testing. AB - The purpose of this study was to develop a method of qualitatively predicting the most likely degradants in a formulation or probing specific drug-excipient interactions in a significantly shorter time frame than the typical 1 month storage testing. In the example studied, accelerated storage testing of a solid dosage form at 50 degrees C, the drug substance SB-243213-A degraded via the formation of two oxidative impurities. These impurities reached a level of 1% PAR after 3 months. Various stressing methods were examined to try to recreate this degradation and in doing so provide a practical and reliable method capable of predicting drug-excipient interactions. The technique developed was able to mimic the 1-month's accelerated degradation in just 1 hr. The method was suitable for automated analysis, capable of multisample stressing, and ideal for use in drug excipient compatibility screening. PMID- 12760563 TI - Study of multiple-component adsorption on the surface of activated carbon using a model system of benzyl alcohol and phenobarbital. AB - The purpose of this work was to characterize the surface of activated carbon and to study the specificity of interactions using multicomponent adsorption. The competition between phenobarbital and benzyl alcohol was studied by conducting multicomponent-adsorption experiments. Benzyl alcohol and phenobarbital were combined to form a bisolute system. The adsorption of the bisolute system from simulated intestinal fluid (without pancreatin) by activated carbon was studied by using the rotating bottle method. The concentrations, both before and after the attainment of equilibrium, were determined with the aid of an HPLC system using a reversed-phase column. The modified competitive Langmuir-like model was fit to the data. A good correlation was obtained between the experimental and the calculated data, which indicates that benzyl alcohol and phenobarbital are competing for the same binding sites. The competition between benzyl alcohol and phenobarbital was not expected, and it suggests that benzyl alcohol is not interacting with the site having the theoretically highest enthalpy of interaction (carbonyl group on the activated carbon surface), due to the blockage of this site by the solvent (water). This unexpected result also indicates that the hydroxyl group is likely to be the most important group when the adsorption occurs from aqueous solution. PMID- 12760564 TI - Design of salbutamol EOP tablets from pharmacokinetics parameters. AB - Salbutamol elementary osmotic pump (EOP) tablets were developed, and fundamental variables affecting their release characteristics were evaluated. The effects of film thickness and compression force on drug release from the tablets containing fixed amount of sodium chloride used as osmogent were evaluated. The core tablets were directly compressed at four compression forces and coated with 3% wt/vol cellulose acetate in acetone to levels of 2%, 3%, and 4% wt/wt. Coated tablets were drilled with CO2 laser beam to form drug delivery orifice of approximately 400 microm in diameter. The drug release was found to follow zero order fashion. The release rate decreased with the increased film thickness and was not affected by the compression force or porosity. The tablets coated to 3% and 4% levels exhibited the release rates within the range calculated from pharmacokinetic data. To illustrate the effect of osmogent content, the tablets were prepared at four osmogent levels and compressed at a constant compression force. The core tablets were coated to a level of 3% wt/wt. The release rate was initially increased with osmogent content and then decreased. At higher osmogent contents, the drug fraction in soluble component was decreased and resulted in the reduction of drug release. In conclusion, film thickness and osmogents played important roles in drug release from EOP tablets. PMID- 12760565 TI - Development of agglomerated directly compressible diluent consisting of brittle and ductile materials. AB - The objective of this investigation was to develop a novel multifunctional coprocessed adjuvant consisting of three known diluents that show different consolidation mechanisms. The method of wet granulation was adopted for the preparation of coprocessed product. Microcrystalline cellulose (MCC) and colloidal silicon dioxide (X1), lactose monohydrate (X2), and dibasic calcium phosphate dihydrate (X3, DCP) were used as independent variables in a simplex lattice design. Croscarmellose sodium was used at 4% level intragranularly in all the batches. The granules (44/120 #) were characterized for angle of repose, bulk density, tapped density, and Carr's index. The tablets of coprocessed adjuvants were characterized for crushing strength, friability, and disintegration time. Multiple linear regression was adopted for evolving refined mathematical models. A checkpoint batch was prepared and evaluated for particle size distribution, moisture uptake, and dilution potential by using nimesulide as a model drug. Microcrystalline cellulose shows poor flowability due to irregular shape and interlocking. Moreover, it loses a part of its compactibility on wet granulation. To attend these problems, a physical blend of 97% microcrystalline cellulose and 3% colloidal silicon dioxide M5 was prepared and used. The blend of MCC and colloidal silicon dioxide showed better flow than that of the original MCC. Hence, it may be easier to mix with lactose and dibasic calcium phosphate. The loss in compactibility of microcrystalline cellulose on wet granulation was also reduced due to presence of colloidal silicon dioxide. As expected, all the batches exhibited acceptable angle of repose (<35 degrees) and quick disintegration (<1 min). Full and refined models for Carr's index and crushing strength were evaluated. Based on the results of grid analysis, a checkpoint (50% MCC, 40% lactose, and 10% DCP) that satisfies both the conditions of Carr's index and crushing strength was selected. The adjuvants absorb very little moisture in the moisture uptake study. The results of dilution potential study reveal that up to 30% nimesulide, a poorly compressible drug, can be incorporated in the coprocessed product. In vitro drug dissolution from capsules containing pure drug powder and compressed tablets was comparable (f2 = 79). The results reveal that the desired product characters can be obtained by varying the quantity of MCC (a ductile material that undergoes plastic deformation), lactose (brittle material with low-fragmentation propensity), and DCP (brittle material with high fragmentation propensity). PMID- 12760566 TI - Pharmacokinetics and distribution of 125I-PLA-b-PEO block copolymers in rats. AB - The PLA-b-PEO block copolymers were studied as potential carriers for anti inflammatory and anticancer drugs. The copolymers were labeled with 125I, and their micelles in physiological saline were prepared by dialysis. Copolymer 1, with Mw = 12,360 (PLA/PEO = 7000/5000), formed particles of about 300 nm in diameter (Rh was 150 nm), whereas copolymer 2, with Mw = 20,470 (6000/14,000), made up micelles of about 72 nm in diameter (Rh was 35.8 nm). Their pharmacokinetic and biodistribution profiles were compared in normal rats and rats with carrageenan-induced inflammation after intravenous application of about 5 mg/kg of each copolymer. Copolymer 2, forming smaller particles, showed longer distribution and elimination half-lives. Both copolymers under study exhibited significantly higher uptake by inflammatory tissue compared with noninflammatory one. The study indicates that PLA-b-PEO copolymers, having different molecular weight of the chains, have similar biological behavior in most organs and tissues. Differences in the uptake by some organs (mainly kidney and bowels) and in activity level in blood at later time intervals were found. Significantly different clearance values are due to different ratios of hydrophobic and hydrophilic chains of the copolymers. PMID- 12760567 TI - A multimechanistic drug release approach in a bead dosage form and in vitro predictions. AB - The objective of this study was to prepare a combination of immediate release, enteric coated, and controlled release (CR) beads and to mathematically model in vitro drug release characteristics of the combination based on the release profiles of individual beads. Uncoated beads were manufactured by using extrusion/spheronization technology. Fluid-bed bottom spraying was used for coating: Eudragit-L-30D for enteric coating and Eudragit-NE-30D for CR coating. In vitro drug release profiles for uncoated and coated beads were each fitted to appropriate mathematical equations. The drug release from the bead combination dosage form was predicted from the individual mathematical models and verified experimentally in vitro. The in vitro dissolution was conducted in 0.1 N HCl for 2 hr and then in buffer (pH 6.5 phosphate, 0.05 M) to mimic in vivo conditions using USP dissolution apparatus I. The results showed that uncoated beads gave similar release profiles in water, acid, and buffer with complete release within 2 hr. The release from CR beads was about 50% at 10 hr and was independent of the dissolution medium. As expected, enteric coated beads showed drug release < 5% at 2 hr in water and acid, whereas the release in buffer was comparable to that of uncoated beads. Exposure of enteric coated beads to acid for 2 hr produced a slower release rate in buffer compared with the release from beads added directly in the buffer. The release characteristics of the three beads can be described by square root and zero-order kinetics. The release characteristics from the combination dosage form were 39%, 69%, and 81% at 1, 4, and 8 hr, respectively. The experimental and predicted profiles agreed to within +/- 6% (residuals at individual data points). Our results suggest that release from the combined multimechanism oral dosage form can be predicted from the performance of individual beads. PMID- 12760568 TI - Exploration of melt granulation technique for the development of coprocessed directly compressible adjuvant containing lactose and microcrystalline cellulose. AB - The objective of the present investigation was to prepare and evaluate lactose and microcrystalline cellulose based, directly compressible adjuvant using melt granulation technique. The percentage of polymer blend (PVP K 30 and PEG 4000; 5, 10, or 15%) and the polymer blend ratio (9:1, 1:1, or 1:9) were selected as independent variables in a 3(2) full factorial design. The lactose and microcrystalline cellulose blend (3:1) was mixed with the meltable binder on a water bath at 90 degrees C. The agglomerates were cooled to 35 degrees C and subsequently passed through 30 mesh. A batch containing 12.5% of the polymer blend containing 1:9 ratio of PVP:PEG was used for further studies. In an another 3(2) full factorial design, disintegrant (crospovidone, croscarmellose sodium, or sodium starch glycolate) and mode of addition of disintegrant (intragranular, extragranular, or combination of intragranular and extragranular) were used as independent variables. The agglomerates were evaluated for percentage fines and Carr's index. Tablets were prepared on a single-punch tablet machine, and they were evaluated for tensile strength, friability, and disintegration time. Regression analysis was carried out to evolve full and refined models. Contour plots are presented for graphical expression of the results. The use of composite index is demonstrated for the selection of an appropriate batch. The disintegration time of tablets reduced from 18 min to 6 min when 6% crospovidone was included in the product. The optimized adjuvant was characterized for particle size distribution, granular friability, Kawakita's and Kuno's equation, and dilution potential study. Turmeric, glycyrrhiza, acetaminophen, and metformin HCl were used as model drugs for the preparation of tablets. The present study underlines the fact that melt granulation technique may be adopted for the development of multifunctional directly compressible adjuvant for use in pharmaceuticals. The advantages of melt granulation technique over the classical wet granulation and spray-drying are presented. PMID- 12760569 TI - Physical stability of micronized powders produced by spray-freezing into liquid (SFL) to enhance the dissolution of an insoluble drug. AB - PURPOSE: The objective of this study was to investigate the physical stability of micronized powders produced by the spray-freezing into liquid (SFL) particle engineeringtechnology. MATERIALS AND METHODS: Danazol was formulated with polyvinyl alcohol (MW 22,000), poloxamer 407, and polyvinylpyrrolidone K-15 to form a cosolvent solution that was SFL processed. The dried micronized SFL powders were sealed in glass vials with desiccant and exposed to 25 degrees C/60% RH for 3 and 6 mo, 40 degrees C/75% RH for 1, 2, 3, and 6 mo, and conditions where the temperature was cycled between -5 and +40 degrees C (6 cycles/24 hr) with constant 75% RH for 1, 2, 3 and 4 wk. The samples were characterized by using Karl-Fisher titration, differential scanning calorimetry, x-ray diffraction, specific surface area, scanning electron microscopy, and dissolution testing. RESULTS: Micronized SFL powders consisting of porous aggregates with small-particle domains were characterized as having high surface areas and consisted of amorphous danazol embedded within a hydrophilic excipient matrix. Karl-Fischer titration revealed no moisture absorption over the duration of the stability studies. Differential scanning calorimetry studies demonstrated high degrees of molecular interactions between danazol, PVA, poloxamer, and PVP. Scanning electron microscopy studies confirmed these interactions, especially those between danazol and poloxamer. These interactions facilitated API dissolution in the aqueous media. Powder surface area remained constant during storage at the various stability conditions, and danazol recrystallization did not occur during the entirety of the stability studies. Micronized SFL powders containing danazol dissolved rapidly and completely within 5 min in aqueous media. No differences were observed in the enhanced dissolution profiles of danazol after exposure to the storage conditions investigated. Physically stable micronized powders produced by the SFL particle engineering technology were produced for the purpose of enhancing the dissolution of an insoluble drug. CONCLUSIONS: The potential of the SFL particle-engineering technology as a micronization technique for enhancing the dissolution of hydrophobic drugs was demonstrated in this study. The robustness of the micronized SFL powders to withstand stressed storage conditions was shown. PMID- 12760570 TI - Buccal delivery of acyclovir from films based on chitosan and polyacrylic acid. AB - The aim of the present work was to investigate the possibility of achieving buccal delivery of a problematic drug, acyclovir, from films based on chitosan hydrochloride (HCS) and polyacrylic acid sodium salt (PAA). At first, the ionic interaction between HCS and PAA in distilled water was investigated by means of rheological and turbidimetric analysis. Films containing 1 mg/cm2 of acyclovir and based on pure HCS and on HCS and PAA mixed in different ratios were prepared by casting technique. The films were subjected to hydration, rheological, mucoadhesion, drug release, "wash away," and permeation/penetration measurements. A commercial cream containing acyclovir and an aqueousacyclovir suspension were used as references. The addition of PAA to HCS produced a decrease in film hydration. Films based on HCS/PAA weight ratio close to interaction productstoichiometry were characterized by higher rigidity and better "wash away" properties with respect to the other films and the reference formulation. The worst mucoadhesive properties were shown by films based on mixing ratios close to interaction product stoichiometry. The addition of PAA to HCS produced a lowering in drug release profile. All the films examined promoted the permeation of acyclovir across porcine cheek epithelium when compared with acyclovir suspension and the commercial cream. The penetration enhancement properties were affected by the mixing ratio of the two polymers. The film based on 1/1.3 HCS/PAA weight ratio, besides possessing the best resilience properties on the mucosa, was also characterized by the highest permeation profile and, therefore, represents a promising formulation for buccal delivery of acyclovir. PMID- 12760571 TI - Complications of iliac crest graft and bone grafting alternatives in foot and ankle surgery. AB - The ability to harvest iliac crest bone is a well-established skill in the surgical armamentarium of the orthopedic surgeon. As with any surgical procedure, this operation has its own set of complications. The surgeon must be aware of these potential problems in an effort to avoid them when possible. Other autologous sites for bone harvest are available to the surgeon, and s/he should be aware of these in terms of location, limitations of use, harvest technique, and potential pitfalls. The foot and ankle surgeon almost always needs less bone graft than our colleagues in spine surgery or joint revision surgery, so these other sites may be more suitable than the iliac crest for obtaining bone graft. Nonautogenous alternatives are becoming increasingly available to the orthopedist as a way to decrease morbidity and operating times. Scranton recently published an article about his success with several different bone substitute products that are used in foot and ankle reconstructive cases. As these options become more varied, it becomes more difficult to know which product to select. Understanding the biology of bone grafting with respect to osteoconduction, osteoinduction, and osteogenesis provides the surgeon with the knowledge that is needed to make an informed choice when selecting a bone grafting option. Before choosing an alternative graft material, the surgeon should also investigate how the graft material has performed in cases similar to his or her patient's needs. In the future, with continued research, the fields of tissue engineering and gene therapy will provide even better options for nonautogenous bone graft material. PMID- 12760572 TI - Salvage of complications of hallux valgus surgery. AB - Complications following hallux valgus surgery can occur and are a frequent source of patient dissatisfaction. The treatment of these complications begins with careful preoperative planning to ensure that the chosen procedure is appropriate for the specific patient. When complications occur, treatment must also be individualized to address the symptoms of the patient. This article presented an overview of the more common complications that are seen following hallux valgus surgery. Nonsurgical and surgical options for treatment were outlined to assist in the management of these complications. PMID- 12760573 TI - Salvage of first metatarsophalangeal joint arthroplasty complications. AB - There are two broad categories of surgical techniques for first metatarsophalangeal joint arthroplasty. Resection arthroplasty is a resection of the base of the proximal phalanx with or without an interposition graft of capsule or adjacent tendon. The second technique involves a partial or complete joint replacement. The primary indication for these procedures is first metatarsophalangeal joint pain caused by arthritic change. The salvage surgical options for failed metatarsophalangeal joint arthoplasty are directed toward decreasing or eliminating the symptoms arising from the complications of these procedures. PMID- 12760574 TI - Morton's neuroma. AB - Morton's neuroma is a common problem. Progress has been made in the understanding of this frequent problem since Morton's original description and treatment. Today, we accept a failure rate of 15% to 20%, even in the best of series. We must ask ourselves if this is good enough. What can we do to achieve an acceptable failure of 5% or less? How can we improve? Only through an honest analysis and discussion can we improve the care that we deliver. PMID- 12760575 TI - Complications of missed or untreated Lisfranc injuries. AB - Injuries to the Lisfranc complex are fairly common. Delayed treatment or missed diagnosis of these injuries can lead to significant complications. Non-operative treatment and salvage surgery can help to relieve sequelae that are associated with tarsometatarsal arthritis following traumatic injury. PMID- 12760576 TI - Osteochondral lesions of the talus. AB - Osteochondral lesions of the talus (OLT) are rare joint disorders. The talus is the third most common location of this disorder, following the knee and elbow joints. OLT represents 4% of all osteochondral lesions in the body. This article discusses the surgical treatment and postoperative rehabilitation of osteochondral lesions of the talus. PMID- 12760577 TI - Stage II flatfoot: what fails and why. AB - Stage II PTT dysfunction is a complex problem that has multiple treatment options. Whether it is treated nonoperatively or operatively, multiple factors affect the success of treatment. The orthopedic literature reports many factors that can lead to failure. Although it is virtually impossible to account for every possible scenario, adhering to sound principles and understanding what can go wrong and why, can help to avoid many of the pitfalls. PMID- 12760578 TI - A review of salvage procedures after failed Achilles tendon repair. AB - The most common complications that are associated with delayed repair of the Achilles tendon are rerupture, wound necrosis, infection, and inability to regain dorsiflexion. Wound complications can be avoided by using meticulous soft-tissue handling techniques, full-thickness flaps when necessary, and minimal pressure to the wound postoperatively. Ankle stiffness may be reduced by ensuring that the repair does not place undue tension on the tendon which requires the ankle to be placed in excessive plantarflexion. Good restoration of function can be obtained by surgical intervention. Regardless of the size of the defect, etiology, or delay in treatment, the salvage procedures described can successfully restore strength and function. It is imperative, however, that patients take an active role in their rehabilitation and remain motivated throughout their course of postoperative therapy. PMID- 12760579 TI - Failed treatment of peroneal tendon injuries. AB - As with most musculoskeletal disorders, understanding the anatomy and function of the peroneus longus and brevis tendons is paramount to the appropriate diagnosis and treatment of their pathologic conditions. This article helps the reader prevent treatment failures by reviewing the anatomy, function, and common treatment options for various disorders of the peroneal tendons. Finally, a discussion of common treatment failures will highlight pitfalls to be avoided. PMID- 12760580 TI - Complications of open reduction and internal fixation of ankle fractures. AB - This article discusses the complications after open reduction and internal fixation of ankle fractures. Complications are classified as perioperative (malreduction, inadequate fixation, and intra-articular penetration of hardware), early postoperative (wound edge dehiscence, necrosis, infection and compartment syndrome), and late (stiffness, distal tibiofibular synostosis, degenerative osteoarthritis, and hardware related complications). Emphasis is placed on preventive measures to avoid such complications. PMID- 12760581 TI - High-risk foot and ankle patients. AB - The surgical treatment of complicated foot and ankle problems requires recognition and appreciation of patient and disease characteristics that may compromise surgical results. Appropriate patient selection is an important factor in obtaining a successful outcome. Modulation and management of these patient characteristics are critical in minimizing complications that are associated with treatment. PMID- 12760582 TI - Failed tarsal tunnel syndrome surgery. AB - Failed tarsal tunnel syndrome surgeries are better prevented than treated. Outcomes for revision procedures are significantly worse than for primaries. Failures should be treated with conservative measures first, then surgery for refractory cases. An adequate release must be ensured, and associatedpathologies must be addressed. One should consider containment procedures for adhesive neuritis and PNS for intraneural or intractable pain. PMID- 12760583 TI - Hypertensive heart failure. PMID- 12760584 TI - Role of infections in atherogenesis. PMID- 12760585 TI - Utility of N-terminal pro-brain natriuretic peptide for the diagnosis of heart failure. AB - BACKGROUND: The goal of this study was to evaluate the utility of plasma N terminal pro-brain natriuretic peptide for the diagnosis of heart failure in patients presenting with shortness of breath. METHODS AND RESULTS: We measured plasma levels of N-terminal pro-brain natriuretic peptide in 119 patients presenting with shortness of breath. The patients were divided into two groups based on the Framingham criteria and echocardiographic results--those with heart failure and those not in heart failure. Plasma levels of N-terminal pro-brain natriuretic peptide were compared in the two groups. The mean N-terminal pro brain natriuretic peptide concentration in patients with heart failure (n=73) was higher than that in those not in heart failure (389+/-148 fmol/ml v. 142+/-54 fmol/ml, p<0.001). N-terminal pro-brain natriuretic peptide values increased significantly as the functional severity of heart failure increased (p<0.001). The mean N-terminal pro-brain natriuretic peptide levels were 261+/-34 fmol/ml for patients in New York Heart Association functional class I, 300+/-161 fmol/ml for patients in New York Heart Association functional class II, 427+/-103 fmol/ml for patients in New York Heart Association functional class III and 528+/-170 fmol/ml for patients in New York Heart Association functional class IV. Using a cut-off value of 200 fmol/ml, the sensitivity of N-terminal pro-brain natriuretic peptide was 97%, specificity was 89% and accuracy for differentiating heart failure from other causes of shortness of breath was 93%. CONCLUSIONS: Our results suggest that N-terminal pro-brain natriuretic peptide can be reliably used for the diagnosis of heart failure in an outpatient setting, and this will improve the ability of clinicians to differentiate patients with shortness of breath due to heart failure from those with other causes of shortness of breath. PMID- 12760586 TI - Usefulness of intravenous metoprolol during positive isoproterenol tilt-table test in the choice of treatment for neurocardiogenic syncope. AB - BACKGROUND: Isoproterenol tilt-table testing provides a diagnosis of neurocardiogenic syncope in patients with syncope or near-syncope. Although acute beta-blockade may prevent the development of syncope during isoproterenol tilt table testing, the use of beta-blockers for chronic prophylaxis may not be effective for some patients who show a positive response to isoproterenol tilt table testing. We evaluated whether the efficacy of intravenous metoprolol in preventing symptoms during repeated tests would be helpful in selecting patients suitable for long-term therapy. METHODS AND RESULTS: We studied 55 patients (35 females, 20 males; mean age 36+/-11 years) who had been chosen from a group referred to our institute with a history of unexplained syncope (> or = 2 syncopal episodes) and a positive response to isoproterenol tilt-table testing. After a positive response to isoproterenol tilt-table testing, 5 mg metoprolol was infused intravenously as a bolus and the test repeated. Thirty-five patients (group 1) showed a positive response again and 20 (group 2) showed a negative response. We started 50 mg metoprolol once a day for patients in group 1 while group 2 was divided into 2 subgroups: the first subgroup (group 2a, 12 patients) was started on 50 mg sertraline or 20 mg paroxetine once a day and the second subgroup (group 2b, 8 patients) was started on 5 mg midodrine orally once a day. Two months later, isoproterenol tilt-table testing was repeated. In group 1, 13 of 35 patients (37%) were positive on isoproterenol tilt-table testing while in group 2, 8 of 20 patients (40%) were positive on isoproterenol tilt-table testing (p not statistically significant). The therapies of the two groups were then interchanged. Two months later (4 months from the beginning of the study), the isoproterenol tilt-table test was repeated. Eleven patients in group 1 (31%) and 6 in group 2 (30%, p not statistically significant) showed a positive response again. CONCLUSIONS: We conclude that acute beta-blockade response to positive isoproterenol tilt-table testing is not a useful predictor for the assessment of chronic prophylaxis for neurocardiogenic syncope. PMID- 12760587 TI - Noninvasive assessment of endothelial dysfunction by brachial artery flow mediated dilatation in prediction of coronary artery disease in Indian subjects. AB - BACKGROUND: A noninvasive technique for testing endothelial function by ultrasound measurement of flow-mediated dilatation has recently generated considerable interest as a marker of atherosclerosis, and in the prediction of clinical coronary events and coronary artery disease. METHODS AND RESULTS: We measured the flow-mediated dilatation of the brachial artery (endothelium dependent vasodilatation) in 136 subjects, with or without evidence of coronary artery disease. Endothelial dysfunction was diagnosed if flow-mediated dilatation was less than 4.5%. Of the 136 subjects (age group 40-70 years) recruited for the study, 94 were males and 42 females. Sixty-eight subjects had evidence of coronary artery disease as diagnosed by documented hospitalization due to myocardial infarction or acute coronary syndrome, proved by coronary angiography when feasible or noninvasive cardiac evaluation. Endothelial dysfunction was detected in 90 subjects (66.2%). Prevalence of coronary artery disease was higher among subjects with endothelial dysfunction compared to those without (57.5% v. 34.7%, p=0.013). Prevalence of endothelial dysfunction was significantly higher among subjects with coronary artery disease as compared to those without coronary artery disease (76.4% v. 55.8%, p=0.012). The present study showed a sensitivity of 76%, specificity of 44%, positive predictive value of 58% and negative predictive value of 65% for endothelial dysfunction in the prediction of coronary artery disease. Multiple regression analysis using coronary artery disease as a dependent variable revealed a statistically significant association with endothelial dysfunction (p=0.033) even after the inclusion of traditional risk factors into the model. CONCLUSIONS: We conclude that endothelial dysfunction shows a strong association with coronary artery disease and can be a useful noninvasive tool for the evaluation of coronary artery disease. PMID- 12760588 TI - Supravalvar aortic stenosis: clinical and hemodynamic profile, and surgical outcome. AB - BACKGROUND: Supravalvar aortic stenosis is the rarest of left ventricular outflow obstructions. Data on this rare entity from India are scarce. METHODS AND RESULTS: We retrospectively analyzed the data of 15 patients (13 males, mean age 15.5+/-10.18 years) with a diagnosis of supravalvar aortic stenosis confirmed by cardiac catheterization. Five patients had morphological features of Williams' syndrome. One patient had diffuse while the rest had discrete type of supravalvar aortic stenosis. Five patients did not have any associated lesions. A 9-year-old male had an ascending aortic aneurysm, and 3 patients had associated peripheral pulmonary artery stenosis. One child had a subaortic ventricular septal defect, and another had severe mitral regurgitation. Twelve patients had electrocardiographic evidence of left ventricular hypertrophy. Three patients had mild aortic valvar stenosis while 2 had aortic regurgitation. Six patients had dilated coronary arteries. Two patients with supravalvar aortic gradients of 20 and 40 mmHg were kept on close follow-up. One patient was not willing to undergo surgery while the other is awaiting surgery. Eleven patients underwent surgical correction. Dacron or pericardial patch aortoplasty was done in all the patients. In addition, one patient each underwent pulmonary artery plasty, ventricular septal defect closure, repair of ascending aortic aneurysm, and mitral valve replacement. The patient with diffuse type of supravalvar aortic stenosis underwent augmentation aortoplasty. Two patients died perioperatively. One was lost to follow-up. Two had moderate residual gradients. The rest of the patients were in New York Heart Association functional class I on follow-up of 6.3+/-4.7 years. CONCLUSIONS: Repair of supravalvar aortic stenosis by single sinus aortoplasty is safe and produces good results. PMID- 12760589 TI - The efficacy and tolerability of sildenafil in patients with moderate-to-severe pulmonary hypertension. AB - BACKGROUND: Pulmonary arterial hypertension is a life-threatening disease for which continuous intravenous infusion of prostacyclin has proved effective. However, it carries the risk of serious complications arising from the complex delivery system. Prostacyclin analogs, endothelin antagonists, and the phosphodiesterase-5 inhibitor sildenafil are emerging promising therapies. This study was aimed at evaluating the utility of oral sildenafil in patients with pulmonary hypertension of varied etiology, poorly controlled on conventional treatment. METHODS AND RESULTS: Ten consecutive patients with pulmonary hypertension, either primary or related to previous left-to-right shunts, thromboembolism, or interstitial lung disease, poorly controlled on conventional therapy such as warfarin, calcium antagonists, digitalis, and diuretics, were included. A thorough clinical, laboratory, and comprehensive echo Doppler evaluation was performed before enrollment in the trial to establish the diagnosis and obtain baseline data. Subjects received sildenafil 25 mg 8 hourly, or a matching placebo for two weeks each, in a randomized, double-blind, crossover design. A run-in period of two weeks was permitted between the two therapies during which patients continued to receive the conventional therapy without any vasodilator. At the end of each therapy period, the patients were evaluated for symptoms, New York Heart Association class, distance covered during the 6 min walk test, rating of modified Borg dyspnea score, and systolic pulmonary artery pressure using echo Doppler. The differences in the above variables at the end of sildenafil and placebo therapies were compared. Nine patients completed the study protocol. Sildenafil, compared to placebo, was associated with improved exercise tolerance as determined by the 6 min walk test (266.67+/-131.45 m v. 170+/-105 m; p<0.005), decrease in modified Borg dyspnea score (3.56+/-1.01 v. 5.11+/-1.45; p<0.01), decrease in Doppler-estimated pulmonary artery systolic pressures (55.33+/-16.52 mmHg v. 75.33+/-19.75 mmHg; p<0.005), improvement in New York Heart Association class (2 patients), and improvement in symptoms. Sildenafil was well tolerated with no untoward effects; further, no significant changes in heart rate or blood pressure occurred during the study period. CONCLUSIONS: Sildenafil improves exercise capacity and symptoms, and decreases pulmonary artery pressures in patients with primary or secondary pulmonary hypertension of varied etiology. PMID- 12760592 TI - Temporary endoepicardial atrioventricular sequential pacing for complete heart block following complex surgery for congenital heart disease. AB - Complete heart block following intracardiac surgical repair for complex congenital heart disease is not uncommon. In the presence of ventricular dysfunction, ventricular pacing alone may not improve the cardiac output. We report the feasibility and efficacy of endoepicardial atrioventricular sequential pacing in a case of postoperative complete heart block. PMID- 12760591 TI - Isolation of the left subclavian artery. AB - Two cases of isolation of the left subclavian artery from the aortic arch are reported for the rarity of this lesion. One patient was diagnosed clinically, the other after angiography. The isolated left subclavian artery was reimplanted in one patient. This rare anomaly has clinical and surgical relevance and should be diagnosed by diligent clinical and angiographic evaluation. PMID- 12760590 TI - Polymorphisms in the apolipoprotein B-100 gene: association with plasma lipid concentration and coronary artery disease. AB - BACKGROUND: The aim of this study was to investigate the association of apolipoprotein B gene polymorphisms with coronary artery disease and lipid levels in Indians. METHODS AND RESULTS: One hundred patients of angiographically proven atherosclerotic coronary artery disease and one hundred age- and sex-matched control subjects (treadmill negative) were included in the study. Serum lipids including cholesterol, triglycerides, high-density lipoprotein, low-density lipoprotein, very low-density lipoprotein, and apolipoprotein B were analyzed. Genomic DNA was extracted and the apolipoprotein B 3' hypervariable region amplified by polymerase chain reaction. Regions carrying Xba1, EcoR1, and Msp1 restriction sites present in the apolipoprotein B gene were amplified and digested separately by the respective enzymes. Restriction fragment length polymorphism analysis showed that EcoR1 with the R+/R+ genotype was significantly more common in patients with coronary artery disease. Overall, the genotypes EcoR1+/+, Msp1+/+, Xba1+/+ and Eco R1+/+ Msp1+/-, Xba1-/- were significantly more common in patients as compared to controls (p<0.05). When gene polymorphisms were compared with lipid abnormalities, the genotypes EcoR1+/+, Xba1-/-, and Msp1+/+ were more frequent in patients with elevated apolipoprotein B and very low density lipoprotein levels. On the other hand, these genotypes were less common in patients with increased total cholesterol and low-density lipoprotein levels. When we studied the individual alleles of the variable number of tandem repeats region, we observed that allele 34 was significantly increased in patients with coronary artery disease as compared to controls. Allele 36 was present with a frequency of 1% in controls while it was totally absent in patients. CONCLUSIONS: This study identifies the apolipoprotein B gene polymorphism associated with coronary artery disease. An association between apolipoprotein B gene polymorphisms and elevated apolipoprotein B and very low-density lipoprotein levels was observed. However, there was no positive association with other elevated lipid levels in North Indians from Uttar Pradesh. PMID- 12760593 TI - Concurrent coronary, bilateral iliac and left renal artery direct stenting. AB - We describe a patient who underwent percutaneous coronary intervention combined with bilateral iliac and left renal artery angioplasty during the same sitting. Stenting of the coronary and peripheral arteries was performed employing the "direct stenting" technique. No complications occurred. The patient was discharged 2 days after the intervention and remains asymptomatic, leading a fully active life during 1 year of follow-up. To our knowledge, unstaged coronary stenting combined with direct stenting of the renal and both common iliac arteries has not been reported previously in India. PMID- 12760594 TI - Catheter ablation of atrial tachycardia using a real-time position management mapping system. AB - Catheter ablation for atrial tachycardia is limited by its low success rate and prolonged procedure time because of difficulties in mapping the site of the tachycardia. A new three-dimensional mapping system, the Cardiac Pathways mapping system, using an ultrasound transducer, has recently become available. We report a case of focal atrial tachycardia ablation with this system. PMID- 12760595 TI - An infant with "dying spells". AB - A 45-day-old infant presented with the unusual and intriguing symptom of episodic crying and loss of consciousness. The infant was discovered to have a vascular compression of the trachea by the innominate artery, almost serendipitously. He was cured of his symptoms by anterior suspension of the innominate artery. PMID- 12760596 TI - Low-molecular-weight heparins in percutaneous coronary interventions. PMID- 12760597 TI - Echocardiographic anatomy of atrial septal defect: "nomenclature of the rims". PMID- 12760598 TI - Cardiovascular Images. Unusual migration of proximally detached pacemaker lead into the coronary sinus. PMID- 12760599 TI - Tissue Doppler echocardiography: a need for review. PMID- 12760600 TI - Hypertension and cancer mortality: is there a place for antioxidant interventions? PMID- 12760601 TI - Protection for a litigious society. PMID- 12760602 TI - Staying abreast of the benefits of nursing. PMID- 12760603 TI - Who will be hired: Stacey or Shakisha? PMID- 12760604 TI - Prescription prices a hard pill to swallow. PMID- 12760605 TI - Implications of menopausal hormonal therapy for African American and Hispanic women. PMID- 12760606 TI - Feeding our children to death: the tragedy of childhood obesity in America. PMID- 12760607 TI - Assessing the impact of direct-to-consumer advertisements on the AA patient: a multisite survey of patients during the office visit. AB - The method by which patients receive health information continues to change in this new age of increased information and technology. Long gone are the days of the mystique surrounding the physician's prescribing of a medication to cure their patients ailment. In recent years, this mystique has been replaced with more informed patients that bring their questions and concerns their doctor in anticipation of open dialogue. At the other end of the spectrum are the patients that come to the visit armed with notes, copies of Web pages, information from their home medical reference books and information heard or seen from the latest advertisements directed to the consumer about a medication for a self diagnosed disease. The majority of physicians have welcomed and encouraged patients to take on more responsibility of their health and appreciate the dialogue. This is preferred to the patient who sits motionless and nods his/her head in blind agreement with the proposed plan of action. Physicians have read the data on patient compliance and medical outcomes and understand that an informed patient is an ally and not an enemy. However, is there a difference between an informed patient that asks questions seeking a more active role in their healthcare versus a demanding patient that believes that their knowledge base is equal to the physician, thereby reducing the physician to a vending machine dispensing requested prescriptions with little insight or deviation from the patient's demands? Fundamentally, we know that physicians agree that our profession dictates that we use all of the medical knowledge at our fingertips, our interpersonal skills and judgment in making recommendations to our patients. We refuse to be reduced to a simple dispenser of desired medications with no regard to the patient and their well being and in fact are incensed with the assumption that our profession has been reduced to such mediocrity. However, we prefer the middle ground with our patients, neither the demanding or the complacent are our preference, we want relationship and believe that this will improve the patient's health status. The question is whether the increased information, dialogue, and patient involvement utilizing the information from new sources are a good thing, a bad thing or simply a reality of medicine's entrance into the information age. Must we encourage our patients to take an active role in their healthcare but place boundaries on the venues by which they receive information? How do patients interact with their physician in the office after exposure to an ad for a prescription medication? How do physicians react to the questions and do we encourage the interaction? What is the effect of the pharmaceutical industry marketing directly to our patients? Do we know and how will we ever fully assess the impact? Although these questions have no succinct answer, the physician and the patient dyad must be the guiding barometer. PMID- 12760608 TI - Incidence of pelvic organ prolapse in Nigerian women. AB - OBJECTIVE: To establish the incidence and types of utero-vaginal prolapse. METHODS: Retrospective medical records analyses of women who were subjected to reconstructive pelvic surgery for various types of pelvic relaxation at the Nnamdi Azikiwe University Teaching Hospital, Nnewi and the University Of Nigeria Teaching Hospital, Enugu, Nigeria was carried out. The study was conducted from January 1996 to December 1999 during which there were 7515 surgical admissions. The inclusion criteria were those women who complained of feeling a mass in the vagina with demonAstrable descent of the anterior and/or posterior and/or apical vaginal walls and/or perineal descent. Excluded were patients who had other symptoms other than utero-vaginal prolapse and those whose grades and sites of prolapse were not determinable from the clinical or surgical notes. Also excluded were patients with nerve injury or disease, connective tissue disorders and neuromuscular diseases. The subjects were divided into two groups. Group I consisted of 54 women (age < or = 40 years), and group II included 105 women (age > or = 40 years). The findings between those two groups were compared with reference to sites, types and degree of prolapse. Also, coexistence of pelvic relaxation and underlying medical conditions were evaluated. RESULTS: A total of 159 subjects out of 492 charts studied met the inclusion criteria for the study. In group I, mean age was 32.839 with a standard deviation (SD) of +/- 6.012 years; and in group II the mean age was 56.543 with a SD of 8.094. Hypertrophic (elongated) cervix was determined in 15 (6.3%) subjects in group I for an incidence of 1.58% per year, cystocele (vaginal anterior wall descent) was present in 21 (8.9%) women for an incidence of 2.2% per year; rectocele (posterior vaginal wall descent) was identified in 15 (6.3%) women for an incidence of 1.58% per year; vaginal cough prolapse (apical descent) was present 21 (8.9%) women for an incidence of 2.2% per year. Perineal descent was absent in this group. In group II, there was no hypertrophic cervix; cystocele was present in 39 (16.5%) cases for an incidence of 4.13% per year; rectocele was identified in 27 (11.4%) women, amounting to an incidence of 2.85% per year; vaginal cough prolapse was present in 36 (15.%) women, an incidence of 3.75% per year; perineal descent was present in 63 (25.6%) women, for an incidence of 6.4% per year. CONCLUSION: (1) The incidence of hypertrophic cervix without any other abnormality amounted to 1.58% per year. This medical entity can present as uterine prolapse and was noted only in group I. (2) The annual incidence for hospital admission with a diagnosis of uterine prolapse was 2.1%. (3) The incidence of cystocele, and rectocele was not statistically different in the two groups; but the incidence of perineal descent and uterine prolapse were significantly more in group II than group I. (4) The etiology of hypertrophic cervix is not known, but it is of importance especially in the childbearing age when it may be related to prolonged pregnancy, cervical dystocia, etc. PMID- 12760610 TI - Handwriting jokes not so funny. PMID- 12760609 TI - Atypical psychotropic medications and their adverse effects: a review for the African-American primary care physician. AB - There are now five new-generation atypical psychiatric medications currently available. As these new treatments have become more common, they have grown to account for a significant percentage of all psychiatric medications prescribed. This is because of their efficacy in the treatment of several psychiatric disorders, ease of administration, and absence of the well-known extrapyramidal adverse effects long-attributed to the standard dopamine blocking anti-psychotic medications. As these medications have become treatments of choice, we have discovered additional information about their respective side effects. Issues such as bone marrow suppression, endocrine abnormalities, and most recently cardiac arrhythmia have produced concern. This paper will address all in an attempt to inform the primary care physician of the most prominent and clinically relevant adverse effects of these agents. A particular focus will address the increasing concern that these new medications can produce hyperglycemia and diabetes mellitus. PMID- 12760611 TI - Environmental factors associated with asthma. AB - Asthma, a disease of attacks and remission, continues to account for substantial morbidity and direct economic costs. Numerous studies--epidemiologic, toxicologic and clinical--present evidence for a broad spectrum of environmental risk factors associated with asthma. This review summarizes current thinking on a subset of these factors. Knowledge of potential environmental determinants of asthma is important to both the patient and healthcare professional in the application of multiple modalities of medical and environmental intervention for management of the development, and exacerbation of this chronic inflammatory disorder of the airways. PMID- 12760612 TI - Primary lymph node gastrinoma: a case report. AB - The Zollinger-Ellison syndrome consists of severe peptic ulceration, acid hypersecretion, and islet tumors known as gastrinomas. The discovery of gastrinomas in unusual locations such as lymph nodes, bones, ovaries, and the liver poses a diagnostic dilemma as to whether the tumor is primary or metastatic. Here we present a case of a primary gastrinoma within a lymph node. PMID- 12760613 TI - Short term evaluation of a rural immunization program in Nigeria. AB - BACKGROUND: Immunization remains the primary strategy in both the control and prevention of common childhood diseases, particularly in the developing world. Immunization and preprimary health care services were commenced in a rural community in Nigeria in 1998, when vaccine coverage for all Expanded Program on Immunization (EPI) diseases (tuberculosis, polio, diphtheria, pertussis, tetanus, measles, and hepatitis B) was considerably low with only 43% of children fully immunized. METHODS: Children aged 0-2 years and living in a rural community were recruited into the study. Data on vaccination history was collected by both vaccination card and maternal history. Three hundred and twenty-seven children were recruited into the study. Study participants were vaccinated for EPI diseases. Hepatitis-B vaccine was administered at birth, and a combined diphtheria and tetanus toxoids, and pertussis whole cell vaccine (DTP) plus hepatitis-B vaccine was administered in a single injection after six weeks. RESULTS AND CONCLUSIONS: Two years after the program was started, immunization coverage rates were 94% for BCG, 88% for DTP (third dose), and 82% for measles. All antigens showed significant improvements from baseline values (p < 0.0001). Eighty four percent of children were fully immunized against all six diseases, compared with 43% at the commencement (p < 0.0001). Hepatitis-B coverage (three doses) was 58%. The vaccination program has significantly improved vaccination coverage and could be a model for under served, non-industrialized communities. PMID- 12760614 TI - Postrecognition of interleaved melodies as an indirect measure of auditory stream formation. AB - Primitive processes involved in auditory stream formation are measured with indirect, objective method. A target melody interleaved with a distractor sequence is followed by a probe melody that was identical to the target or differed by 2 notes. Listeners decided whether the probe melody was present or not in the composite sequence. Interleaved melody recognition is not possible when distractor sequences have the same mean frequency and maximum contour crossover with target melodies. Performance increases with mean frequency separation and timbral dissimilarity and is unaffected by the duration of the silent interval between composite sequence and probe melody. The relation between this indirect task measuring the interleaved melody recognition boundary and direct judgments measuring the fission boundary is discussed. PMID- 12760615 TI - Vanishing dual-task interference after practice: has the bottleneck been eliminated or is it merely latent? AB - Practice can, in some cases, largely eliminate measured dual-task interference. Does this absence of interference indicate the absence of a processing bottleneck (defined as an inability to carry out certain stages in parallel)? The authors show that a bottleneck need not produce any observable interference, provided that there is no temporal overlap in the demand for bottleneck stages on the 2 tasks. Such a "latent" bottleneck is especially likely after practice, when central stages are short. The authors provide new evidence that a latent bottleneck occurred for a participant who produced no interference in M. Van Selst, E. Ruthruff, and J. C. Johnston (1999). These findings demonstrate that the absence of dual-task interference does not necessarily indicate the absence of a processing bottleneck. PMID- 12760616 TI - Phase attraction in sensorimotor synchronization with auditory sequences: effects of single and periodic distractors on synchronization accuracy. AB - Four experiments showed that both single and periodic distractor tones affected the timing of finger taps produced in synchrony with an isochronous auditory target sequence. Single distractors had only small effects, but periodic distractors occurring at various fixed or changing phase relationships exerted strong phase attraction. The attraction was asymmetric, being stronger when distractors preceded target tones than when they lagged behind. A large pitch difference between target and distractor tones (20 vs. 3 semitones) did not reduce phase attraction substantially, although in the case of continuously changing phase relationships it did prevent complete capture of the taps by the distractors. The results support the hypothesis that phase attraction is an automatic process that is sensitive primarily to event onsets. PMID- 12760617 TI - Representing spatial location and layout from sparse kinesthetic contacts. AB - Participants' fingers were guided to 2 locations on a table for 3 s, then back to the start. They reported distances and angles between the locations by (a) replacing 1 or 2 fingers, (b) translating the contacted configuration, or (c) estimating distance or angle alone. Distance error increased across these conditions. Angular error increased when the angular reference axis was rotated before the response. Replacing 1 finger was impaired by a change in posture from exposure to test. The results suggest a kinesthetic representation is used to replace the fingers, but to estimate distance and angle at new locations, a configural representation is computed. This presentation is oriented within an extrinsic reference frame and maintains shape more accurately than scale. PMID- 12760618 TI - Mutual interpersonal postural constraints are involved in cooperative conversation. AB - The research was designed to evaluate interpersonal coordination during conversation with a new measurement tool. The experiment uses an analysis based on recurrence strategies, known as cross recurrence quantification, to evaluate the shared activity between 2 postural time series in reconstructed phase space. Pairs of participants were found to share more locations in phase space (greater recurrence) in conditions where they were conversing with one another to solve a puzzle task than in conditions in which they convened with others. The trajectories of pairs of participants also showed less divergence when they conversed with each other than when they conversed with others well. This is offered as objective evidence of interpersonal coordination of postural sway in the context of a cooperative verbal task. PMID- 12760619 TI - Change deafness: the inability to detect changes between two voices. AB - A shadowing task was used to demonstrate an auditory analogue of change blindness (the failure to detect a change in a visual scene), namely change deafness. Participants repeated words varying in lexical difficulty. Halfway through the word list, either the same or a different talker presented the words to participants. At least 40% of the participants failed to detect the change in talker. More interesting is that differences in shadowing times were found as a function of change detection. Alternative possibilities to the change detection phenomenon were ruled out. The results of these experiments suggest that the allocation of attention may influence the detection of changes as well as the processing of spoken words in complex ways. PMID- 12760620 TI - Behavioral dynamics of steering, obstacle avoidance, and route selection. AB - The authors investigated the dynamics of steering and obstacle avoidance, with the aim of predicting routes through complex scenes. Participants walked in a virtual environment toward a goal (Experiment 1) and around an obstacle (Experiment 2) whose initial angle and distance varied. Goals and obstacles behave as attractors and repellers of heading, respectively, whose strengths depend on distance. The observed behavior was modeled as a dynamical system in which angular acceleration is a function of goal and obstacle angle and distance. By linearly combining terms for goals and obstacles, one could predict whether participants adopt a route to the left or right of an obstacle to reach a go (Experiment 3). Route selection may emerge from on-line steering dynamics, making explicit path planning unnecessary. PMID- 12760621 TI - Controlling steering and judging heading: retinal flow, visual direction, and extraretinal information. AB - The contribution of retinal flow (RF), extraretinal (ER), and egocentric visual direction (VD) information in locomotor control was explored. First, the recovery of heading from RF was examined when ER information was manipulated; results confirmed that ER signals affect heading judgments. Then the task was translated to steering curved paths, and the availability and veracity of VD were manipulated with either degraded or systematically biased RF. Large steering errors resulted from selective manipulation of RF and VD, providing strong evidence for the combination of RF, ER, and VD. The relative weighting applied to RF and VD was estimated. A point-attractor model is proposed that combines redundant sources of information for robust locomotor control with flexible trajectory planning through active gaze. PMID- 12760622 TI - Task goals and change in dynamical degrees of freedom with motor learning. AB - In this article, the authors examined the hypothesis that the direction of the change (increase or decrease) in the dynamical degrees of freedom (dimension) regulated as a function of motor learning is task-dependent. Adult participants learned 1 of 2 isometric force-production tasks (Experiment 1: constant force output; Experiment 2: sinusoidal force output) over 5 days of practice and a 6th day with augmented information withdrawal. The results showed that over practice, the task goal induced either an increase (Experiment 1) or a decrease (Experiment 2) in the dimension of force output as performance error was reduced. These findings support the proposition that the observed increase or decrease in dimension with learning is dependent on both the intrinsic dynamics of the system and the short-term change required to realize the task goal. PMID- 12760623 TI - Failures of retrieval and comparison constrain change detection in natural scenes. AB - In a change detection paradigm, a target object in a natural scene either rotated in depth, was replaced by another object token, or remained the same. Change detection performance was reliably higher when a target postcue allowed participants to restrict retrieval and comparison processes to the target object (Experiment 1). Change detection performance remained excellent when the target object was not attended at change (Experiment 2) and when a concurrent verbal working memory load minimized the possibility of verbal encoding (Experiment 3). Together, these data demonstrate that visual representations accumulate in memory from attended objects as the eyes and attention are oriented within a scene and that change blindness derives, at least in part, from retrieval and comparison failure. PMID- 12760624 TI - The visual control of reaching and grasping: binocular disparity and motion parallax. AB - The primary visual sources of depth and size information are binocular cues and motion parallax. Here, the authors determine the efficacy of these cues to control prehension by presenting them in isolation from other visual cues. When only binocular cues were available, reaches showed normal scaling of the transport and grasp components with object distance and size. However, when only motion parallax was available, only the transport component scaled reliably. No additional increase in scaling was found when both cues were available simultaneously. Therefore, although equivalent information is available from binocular and motion parallax information, the latter may be of relatively limited use for the control of the grasp. Binocular disparity appears selectively important for the control of the grasp. PMID- 12760625 TI - Understanding other people's actions: intention and attention. AB - This study investigated the extent to which observation of an action performed by a human actor or a robotic arm may kinematically prime the performance of an observer subsequently required to perform a similar action. In Experiment 1, an actor reached for a target presented in isolation or flanked by a distractor object. Subsequently, an observer was required to perform a similar action toward the target object, but always in the absence of the distractor. The kinematics of both the human actor and the observer were affected by the presence of the distractor. Unexpectedly, similar effects were found in the observer's kinematics during the trials in which the actor was seated in front of the observer but no action was demonstrated (catch trials). Results from 4 subsequent experiments suggest that the motor intentions of the actor can be inferred by monitoring his or her gaze. To support this conclusion, results are discussed in terms of recent work spanning many disciplines involved in combining gaze direction and body movements. PMID- 12760626 TI - Can blindness to response-compatible stimuli be observed in the absence of a response? AB - Blindness to response-compatible stimuli is the finding that targets are identified less accurately when presented during the planning or execution of a congruent response (e.g., right arrow presented during a right keypress) versus an incongruent response (e.g., right arrow presented during a left keypress). Accounts of this effect suggest the planning and execution of a response are critical to its observation. Five experiments investigated whether a blindness effect would be observed in the absence of a planned response. Results suggest that a planned response is not necessary to observe a content-specific blindness effect and that the blindness effect may actually comprise both an action-related component and a symbolic component that is distinct from the action-planning system. PMID- 12760627 TI - Utterance format affects phonological priming in the picture-word task: implications for models of phonological encoding in speech production. AB - Picture-word experiments investigating the production of multiword utterances with distractors that are phonologically related to words in noninitial position have yielded inconsistent results, ranging from facilitation to inhibition. A comparison of these studies is complicated by differences in detail. In parallel to the empirical inconsistencies, different theoretical accounts of phonological encoding in speech production have been provided. In the present article, the authors propose a unitary account, which can in principle account for facilitation, null effects, and inhibition. It assumes a graded activation pattern of the elements within the scope of phonological advance planning. The account is tested in an experiment varying utterance format while keeping all other aspects constant. The results are consistent with the proposed unitary account. PMID- 12760628 TI - Switching between tasks of unequal familiarity: the role of stimulus-attribute and response-set selection. AB - It has been reported that it is harder to switch to a strong, well-practiced task from a weaker, less-practiced task than vice versa. Three experiments replicated this surprising asymmetry and investigated how it is affected by a reduction in interference between tasks. Experiment 1 progressively delayed the onset of the stimulus attribute associated with the stronger task. Experiments 2 and 3 separated the response sets of the tasks. Both manipulations reduced, without eliminating, interference of the stronger with the weaker task but reversed the asymmetry of switch costs, resulting in a larger cost of switching to the weaker task. The results are interpreted in terms of a model of the interactions between control input, task strength, and task priming. PMID- 12760629 TI - The costs and benefits of tonal centers for chord processing. AB - Harmonic priming studies have shown that a musical context, with its established tonal center, influences target chord processing. This study investigated costs and benefits of priming tonal centers for target processing by adding a baseline condition (sequences without a specific tonal center). Results confirmed harmonic priming, with faster processing for related than for unrelated and less related targets (tonic chord, out-of-key chord, subdominant chord). Comparing targets in baseline contexts with targets in sequences with well-established tonal centers revealed a benefit of processing for related targets but a cost of processing for unrelated and less related targets. Findings are discussed in terns of tonal knowledge activation and suggest that an activated tonal center gives rise to strong expectations for the tonic. PMID- 12760630 TI - Changing your mind: on the contributions of top-down and bottom-up guidance in visual search for feature singletons. AB - Observers, searching for targets among distractor items, guide attention with a mix of top-down information--based on observers' knowledge--and bottom-up information--stimulus-based and largely independent of that knowledge. There are 2 types of top-down guidance: explicit information (e.g., verbal description) and implicit priming by preceding targets (top-down because it implies knowledge of previous searches). Experiments 1 and 2 separate bottom-up and top-down contributions to singleton search. Experiment 3 shows that priming effects are based more strongly on target than on distractor identity. Experiments 4 and 5 show that more difficult search for one type of target (color) can impair search for other types (size, orientation). Experiment 6 shows that priming guides attention and does not just modulate response. PMID- 12760631 TI - A comparative study of biochemical and immunological properties of triosephosphate isomerase from Taenia solium and Sus scrofa. AB - We produced the Taenia solium triosephosphate isomerase (TPI) in Escherichia coli and compared its biochemical and immunological properties with those of the commercial TPI from Sus scrofa. Taenia solium TPI is a homodimer composed of two 27-kDa monomers, with a specific activity of 5,683 U/mg and a Km value of 0.758, and S. scrofa TPI is also dimeric with similar monomeric molecular weight, specific activity of 4,227 U/mg, and a Km value of 0.51. The catalytic parameters for the isomerization of glyceraldehyde 3-phosphate, affinity between TPI monomers, and kinetic thermal denaturation and inactivation were similar for both enzymes. Anti-T. solium TPI antibodies cross-react weakly with Schistosoma mansoni TPI but do not cross-react with S. scrofa, human, or protozoan TPIs. These antibodies inhibited T. solium TPI activity but did not affect S. scrofa enzymatic activity. Immunizations with 1 microg of the T. solium TPI reduced 52% of cysticerci in a mouse-Taenia crassiceps model 1 mo after challenge. Our findings show that T. solium and S. scrofa TPIs possess similar biochemical and enzymatic properties but do not share immunological properties because anti-T. solium TPI antibodies did not recognize S. scrofa TPI. Inhibition of enzyme activity by anti-TPI antibodies suggests that they can be used as inhibitors of the enzyme. PMID- 12760632 TI - Circulation of parasites among fishes from lakes in the Caribou Mountains, Alberta, Canada. AB - Circulation of 24 macroparasite species among 12 species of fish was evaluated within samples of hosts collected from 9 lakes on an isolated plateau in northern Alberta, Canada. Twenty-seven parasite taxa (24 species plus the larval stages of Triaenophorus crassus, T. stizostedionis, and Raphidascaris acus) had the potential to be circulated among hosts. Sixteen parasite taxa were recovered from a single host species within a lake. Of the 11 remaining nonspecialist taxa, 4 were larval stages that matured in fish or birds and 7 were adults. Eight of the 11 cases of circulation among hosts involved lake whitefish, and this host was involved in the transmission of 5 species to piscivorous fishes. Despite evidence for the circulation of 7 taxa among the 4 species of sympatric Salmonidae, 60-99% of all worms were recovered from just 1 species of host. These results indicated that approximately 60% of the parasite taxa that infected fish in these lakes were absolute host specialists. The remaining 40% of parasite taxa had restricted host ranges, with most examples of parasite circulation limited to the 2 species of sympatric coregonid. PMID- 12760633 TI - Social, political, and economic factors responsible for the reemergence of trichinellosis in Serbia: a case study. AB - Over the past decade, eastern Europe has experienced a resurgence of trichinellosis. A recent outbreak in Serbia, Yugoslavia, from December 2001 to January 2002, involving 309 people, revealed many of the causes for this reemergence. Epidemiological investigations indicate that the immediate cause of the recent outbreak was the consumption of smoked sausages produced by a small slaughterhouse or meat processor. However, failure of in-house meat inspection procedures and quality assurance as well as oversight by official veterinary control were also responsible. Further analysis of this breakdown in the food safety net revealed additional general factors that have yielded a seriously deficient veterinary control system, and these are factors that are relevant to the problems experienced throughout eastern Europe and other regions. The recent civil war that led to the breakup of the former Federation of Yugoslavia resulted in severe economic and demographic changes, including high inflation and external economic sanctions. This led to (1) the loss of large numbers of experienced veterinary control officers and their replacement with inexperienced personnel, (2) a change in the swine industry with reduction in the number of large establishments with in-house inspection and replacement with more than 1,000 small abattoirs, too small to afford full-time in-house inspection, and (3) an increase in smallholder pig farming with reduced government oversight to ensure high standards in pig-rearing practices (infection risk management). The consequences of these events have been a 300% increase in Serbian pig infection and a concomittant large increase in human outbreaks. Before 1990, swine trichinellosis in Serbia was confined to 4 small districts, but today about one third of the Republic is considered endemic for trichinellosis. The reemergence of trichinellosis in Serbia illustrates the ability of this zoonosis to "leak" through a poorly maintained food safety barrier and the vulnerability of effective veterinary control to national and international events. PMID- 12760634 TI - Immunity, antigenic heterogeneity, and aggregation of helminth parasites. AB - Empirical studies of helminth parasites reveal that the distribution of parasite burdens in their host populations is highly aggregated. This aggregation is fundamental to the ecology and epidemiology of helminth parasites. Results from a stochastic model predict that aggregation of helminth parasites is inversely related to the intensity of host immunity. Aggregation also decreases with antigenic heterogeneity and increases with heterogeneity in transmissibility among parasite strains. It is also found that the degree of aggregation is greater when immunity affects parasite fecundity than when immunity acts on host susceptibility. Potential relevance of this result for assessing the influence of vaccines that target either host susceptibility or parasite fecundity on the level of aggregation and consequent effects on drug resistance and disease prevalence are discussed. PMID- 12760635 TI - Infection of Sphyrion laevigatum (Copepoda: Sphyriidae) on Genypterus blacodes (Pisces: Ophidiidae) from the Falkland Islands, South Atlantic. AB - The degree of parasitism by the parasitic copepod Sphyrion laevigatum on kingclip Genypterus blacodes in the Falkland Islands was investigated. In a sample of 719 kingclip, ranging in size from 35 to 145 cm total length, the prevalence of infection was 49%. The number of parasites per fish ranged from 0 to 24, with only 2% of the sample having more than 5 parasites. There was a significant increase in mean abundance of parasites with increasing fish age and length (P < 0.01 and P < 0.05, respectively). In addition, there was a significant difference in the prevalence in 1 of 3 geographical areas for the largest size class. PMID- 12760636 TI - Interspecific parasite exchange in a mixed colony of birds. AB - Studies of avian host-parasite interactions rarely include consequences of relationships among hosts for either the host or parasite species. In this study, we examine the ectoparasitic burden of adult and nestling European bee-eaters (Merops apiaster) and rock sparrows (Petronia petronia) in a mixed colony. We found that (1) each bird species had its own species of lice; (2) hematophagous mites parasitized both adults and nestlings of both bird species; (3) Carnus hemapterus, a common parasite of nestling bee-eaters, also infested rock sparrow nestlings, a species not previously described as a host for this dipteran; and (4) whereas C. hemapterus did not show high host specificity within the colony, the emergence of adult flies was synchronized with the start of hatching in bee eater nests. We suggest that coexistence of these 2 bird species results in parasite exchange, bee-eaters obtaining mites from sparrows and sparrows becoming infested by C. hemapterus. Differences in the detrimental effects of parasite transfer for each host species may result in a process of apparent competition mediated by shared parasites. Interspecific parasite exchange is an important aspect of host-parasite relationships in mixed colonies, which requires further attention. PMID- 12760637 TI - Effect of cystacanth body size on adult success. AB - Laboratory-reared cystacanths of Leptorhynchoides thecatus (Acanthocephala: Rhadinorhynchidae) were used to study the effect of cystacanth size on adult success and the factors that influence cystacanth size within the intermediate host. To assess how host size and intensity of infection influence cystacanth size, infected amphipods (Hyalella azteca) were measured, and sex, length, and width of cystacanths were determined. After a subset of cystacanths was measured, small- and large-size classes of cystacanths were designated. To determine how cystacanth size relates to adult size, green sunfish (Lepomis cyanellus) were fed 10 large or small cystacanths. Fish were dissected 6 wk after infection, and worms were removed. After adult worms were permanently mounted on slides, their length and width were measured. Intensity of infection and amphipod size significantly influenced cystacanth size in that large amphipods harbored larger cystacanths than did small amphipods and heavy infections produced smaller cystacanths than did light infections. Adult worms from the small and large cystacanth-size classes showed no significant difference in size; however, large cystacanths had a significantly higher establishment and survival than did small cystacanths: 40% of large worms and 14% of small worms were recovered. The results of this study indicate that host size and host sharing influence cystacanth size and that cystacanth size is an important factor in determining adult success. PMID- 12760638 TI - Plasmodium yoelii: semiquantitative analyses of circumsporozoite protein gene expression during the sporogonic development of P. y. yoelii and P. y. nigeriensis in the mosquito vector Anopheles stephensi. AB - Malaria infection in the mosquito vector can be modulated by the vertebrate host, mosquito factors, and interactions between different parasite populations. Modulation of parasite development can be assessed through the study of gene expression. The present report describes a specific, sensitive, and nonradioactive method that permits assessment of parasite load and quantification of circumsporozoite protein gene expression during the sporogonic stages of Plasmodium yoelii yoelii and P. y. nigeriensis. A decrease in parasite load was observed when comparing DNA of oocysts on day 7 postinfection with that of oocysts and sporozoites on day 19. On day 7, parasites (oocysts) showed a marked increase of circumsporozoite protein expression when compared with that (sporozoites and oocysts) on day 19. The method developed in this work can be a valuable tool to understand parasite interaction mechanisms that are involved in mosquito malaria infections. PMID- 12760640 TI - Role of murine intestinal intraepithelial lymphocytes and lamina propria lymphocytes against primary and challenge infections with Cryptosporidium parvum. AB - To investigate the role of intestinal lamina propria lymphocytes (LPL) and intraepithelial lymphocytes (IEL) in controlling Cryptosporidium parvum infection, changes in their phenotypes and functional properties were studied after induction of primary and challenge infections in immunocompetent mice. As shown by oocyst-shedding patterns, the challenge-infected group recovered more rapidly from infection than did the primary-infected group. In LPL, proportions of activated CD4+, CD25+, IgG1+, IgA+, and CD4+/IFN-gamma+ cells increased significantly in the primary-infected group compared with controls. In the challenge-infected group, proportions of these cells decreased. The antigen specific IgA level was elevated significantly among LPL of both primary- and challenge-infected groups. Among IEL, proportions of activated CD8+, T cell receptor (TCR) gammadelta+, and CD8+/TCR gammadelta+ cells increased significantly in the challenge-infected group compared with controls and the primary-infected group; their cytotoxicity also was enhanced. However, the proportion of IEL expressing Th1 cytokines was lower than that among LPL in both infected groups. The results suggest that LPL play a more important role in protection against a primary infection with C. parvum, through the production of IFN-gamma and IgA, whereas IEL are more involved in protection against a challenge infection, through enhanced cytotoxicity. PMID- 12760639 TI - The Schistosoma mansoni gene index: gene discovery and biology by reconstruction and analysis of expressed gene sequences. AB - Expressed sequence tag (EST) sequencing and analysis is a primary research tool to identify and characterize the Schistosoma mansoni transcriptome. As part of our gene discovery effort, a total of 5,793 ESTs have been generated from clones selected randomly from complementary DNA (cDNA) libraries constructed from male and female adult worms. Assembly analysis of all the 16,813 public S. mansoni ESTs has identified 1,920 distinct tentative consensus sequences (TCs) and 5,571 nonoverlapping ESTs (singletons). Of these, 376 TCs (20%) and 1,449 singletons (26%) are unique to the SUNY/TIGR sequencing effort. Tentative consensus sequences and singletons were distributed into various categories of biological roles associated with cell structure, metabolism, protein fate, signal transduction, transcription, protein synthesis, transporters, and cell growth. The TCs and singletons represent transcripts that can be used as a resource for functional annotation of genomic sequence data, comparative sequence analysis, and cDNA clone selection for microarray projects. The utility of EST analysis is demonstrated by identifying new protease genes, which may be involved in hemoglobin degradation. PMID- 12760642 TI - Wolbachia endosymbionts in fleas (Siphonaptera). AB - Intracellular endosymbionts, Wolbachia spp., have been reported in many different orders of insects and in nematodes but not previously in fleas. This is the first conclusive report of Wolbachia spp. within members of the Siphonaptera. Using nested polymerase chain reaction (PCR) targeting of the 16S ribosomal RNA gene, we screened for Wolbachia spp. in fleas collected from 3 counties in Georgia and 1 in New York. The prevalence of Wolbachia spp. detected varied among the 6 different species screened: 21% in the cat flea Ctenocephalides felis (n = 604), 7% in the dog flea C. canis (n = 28), 25% in Polygenus gwyni (n = 8), 80% in Orchopeas howardi (n = 15), 94% in Pulex simulans (n = 255), and 24% in the sticktight flea Echidnophaga gallinacea (n = 101). Wolbachia spp. infection in fleas was confirmed by sequencing positive PCR products, comparing sequenced 16S ribosomal DNA (rDNA) with Wolbachia spp. sequences in GenBank using BLAST search, and subjecting sequence data to phylogenetic analysis. For further confirmation, 16S rDNA-positive samples were reamplified using the wsp gene. PMID- 12760641 TI - Recombinant bovine herpesvirus-1 expressing p23 protein of Cryptosporidium parvum induces neutralizing antibodies in rabbits. AB - In order to develop a vaccine against cryptosporidiosis in cattle, we constructed a recombinant bovine herpesvirus-1 (BHV-1) expressing an immunodominant surface protein, p23, of Cryptosporidium parvum sporozoites. In the recombinant virus, the p23 gene under the control of a CAG promoter and a gene coding for an enhanced green fluorescent protein were integrated into the gG gene of BHV-1. Despite a low frequency of homologous recombination, cloning of the recombinants was easy because of the specific fluorescence of the plaques formed by recombinants. These plaques were among the plaques of the nonfluorescent parental virus. All clones selected for fluorescence also contained the p23 gene. In MDBK cells infected with the recombinant BHV-1, the antibody against the p23 protein recognized the p23 protein as an approximately 23-kDa specific band in Western blotting analysis. Rabbits immunized with the recombinant produced IgG against the p23 protein. It was also demonstrated that the sera of immunized rabbits reduced infection of C. parvum sporozoites in HCT-8 cells. The serum of an immunized rabbit reduced infection compared with the normal rabbit serum control. These results indicate that the recombinant BHV-1 induces neutralizing antibodies in rabbits. PMID- 12760643 TI - Epizootiology of Eustrongylides ignotus in Florida: transmission and development of larvae in intermediate hosts. AB - Under laboratory conditions, 2 modes of transmission of Eustrongylides ignotus (Nematoda: Dioctophymatoidea) to fish were identified. Eastern mosquitofish (Gambusia holbrooki) became infected after ingestion of either eggs of E. ignotus containing first-stage larvae or aquatic oligochaetes (Limnodrilus hoffmeisteri) containing third-stage larvae of E. ignotus. After removal from the uterus of gravid E. ignotus females and incubation for 17-28 days, depending on temperature, it was found that parasite eggs contained first-stage larvae that were infective to fish and oligochaetes. Larvae developed to the third stage in oligochaetes and were infective to fish 35-77 days postinfection (PI) and when fed to fish, developed to the fourth stage between 127 and 184 days PI. Eggs containing first-stage larvae fed directly to fish developed to the fourth stage between 84 and 105 days PI. The amount of time for development from the undifferentiated egg to the fourth-stage larva was 78-156 days shorter when fish ingested eggs containing first-stage larvae than when fish ingested oligochaetes containing third-stage larvae. Three species of large piscivorous fish, including black crappie (Pomoxis nigromaculatus), largemouth bass (Micropterus salmoides), and warmouth (Lepomis gulosus), were fed mosquitofish containing fourth-stage larvae. At necropsy, live E. ignotus larvae were recovered from all 3 species. Several fish had multiple infections after ingesting > 1 larva, indicating that bioaccumulation of the parasite in the food chain may occur. PMID- 12760645 TI - Prevalence of strongyle nematodes in naturally infected ponies of different ages and during different seasons of the year in Louisiana. AB - One hundred and seventeen ponies were surveyed for the seasonal prevalence of strongyloid parasites, particularly cyathostomes, and for host- and age-related differences in these infections. For 56 ponies, all stages of the cyathostome life cycle, both mucosal and luminal, were enumerated. Total numbers of cyathostomes and percentage of developing larvae (DL) encysted in the mucosa remained constant in all 4 seasons of the year, whereas a significant increase in the percentage of adults in the cyathostome population occurred in fall. In yearling ponies, encysted early third-stage larvae constituted a significantly lower percentage of the cyathostome population, and DL and adults formed a significantly higher percentage, compared with those stages in older ponies, 2-5 yr of age. More species of cyathostomes were present in yearling ponies than in older ponies. Significant differences occurred in fecal egg counts at different seasons of the year, even though adult cyathostome and large strongyle numbers remained constant. Twenty-four species of cyathostomes were found year-round, and 2 rare species were found in only 2 or 3 seasons of the year. Prevalences for these 24 species were not significantly different during any season, although 5 species had significant differences in intensity levels of infection in certain seasons of the year. Three species of large strongyle (strongylinae) adults had significant seasonal variations in intensities. These were Strongylus edentatus and S. vulgaris, which occurred in significantly higher numbers in summer and fall, and Triodontophorus brevicauda, which was more numerous in spring and summer. Fourth- and fifth-stage larvae of S. vulgaris recovered from the mesenteric vasculature were significantly more numerous in winter and spring than in other seasons. PMID- 12760644 TI - The nematode fauna of long-nosed mice Oxymycterus spp. from the Bolivian Yungas. AB - During a long-term survey of the parasites of mammals from all over Bolivia, 44 individuals of Oxymycterus inca and 6 of Oxymycterus paramensis were collected and examined for parasites from the foothills and Eastern Cordillera (Los Yungas) of the Andes of Bolivia. Three species of nematode were found including, from the cecum, a previously unknown genus and species of pinworm, Caroloxyuris boliviensis n. gen, n. sp., aspidoderids representing Nematomystes rodentophilus, and from the stomach, Protospirura numidica criceticola. These helminths occurred in prevalences of 25, 9, and 14%, respectively, in O. paramensis. Caroloxyuris boliviensis resembles species included in the genus Syphacia in the structure of the cephalic mask and copulatory organs, and the extension of lateral alae. However, males of this species possess only 2 mamelons on the ventral surface of the body. Caroloxyuris can be recognized as being distinct from both Helminthoxys and Rauschtineria in the ornamentation of the mamelons and the shape of cephalic mask. A redescription of N. rodentophilus is provided, and Nematomystes scapteromi is transferred from Ansiruptodera to Nematomystes. PMID- 12760647 TI - Reconstructing the spread of Dirofilaria immitis in California coyotes. AB - Dirofilaria immitis is a filarial nematode parasite that is currently widely enzootic in dog and coyote (Canis latrans) populations of California. Weak historical evidence suggests that the initial focus of D. immitis in California occurred 3 decades ago in the Sierra Nevada foothills (SNF) and spread to other parts of California thereafter. However, this hypothesis is difficult to evaluate because of the lack of epidemiological studies on heartworm in California before 1970. We investigated this hypothesis by comparing D. immitis prevalence in coyotes between initial (1975-1985) and current (2000-2002) surveys in the SNF and 2 coastal regions. In the SNF, prevalence of heartworm was not significantly different between initial (35%, n = 169) and current (42%, n = 60) surveys (P = 0.17), suggesting the existence of a stable enzootic focus in the initial survey period. In contrast, current prevalence was 4 times higher than initial prevalence in the northern Coast Range foothills (44 vs. 10%; n = 119, 107; P < 0.001) and in the south San Francisco Bay foothills (32 vs. 8%; n = 31, 59; P = 0.005), suggesting that initial surveys were made during the early stages of colonization. Dirofilaria immitis prevalence, intensity, and abundance was similar in a coastal location in Mendocino County between 1994-1996 and 1999 2002, suggesting some degree of stability in this enzootic focus. Collectively, these findings support the hypothesis that D. immitis established itself initially in California coyotes living in the SNF and subsequently expanded its range of enzootic foci in central California. PMID- 12760646 TI - A survey of the gastrointestinal nematodes of Spanish ibex (Capra pyrenaica) in a high mountain habitat. AB - We analyzed the content of the abomasum (n = 79) and small intestine (n = 83) of Spanish ibex from Sierra Nevada Natural Park, southern Spain. Fifteen species of trichostrongylid nematodes were identified, 4 of which were found for the first time in this host, i.e., Nematodirus fillicollis, N. oiratianus, Ostertagia lyrata, and O. ostertagi. Teladorsagia circumcincta and Marshallagia marshalli were the most abundant abomasal species, whereas N. abnormalis, N. davtiani, and N. oiratianus were dominant in the small intestine. Counts of both abomasal and intestinal nematodes were generally low (year-round-median = 292 and 94 worms, respectively), and significantly lower numbers of M. marshalli, N. davtiani, and N. oiratianus were found in summer. No sex-related differences in helminth abundance were found, but young ibex harbored significantly more N. davtiani and N. oiratianus than adults. The presence of scabies was not related to increased nematode counts. PMID- 12760648 TI - Seasonal dynamics of the helminths of bluegill (Lepomis macrochirus) in a subtropical region. AB - Seasonal prevalence and abundance of the helminths of bluegill sunfish in a Louisiana oligohaline bayou were measured by a survey and a field "live-box" experiment. The survey took place from the spring of 1997 to the summer of 1998 and examined fish that were <7 cm. Three trematode species, Phagicola nana (Heterophyidae), Ascocotyle tenuicollis (Heterophyidae), and Posthodiplostomum minimum (Diplostomatidae), and 2 nematode species, Camallanus oxycephalus (Camallanidae) and Spinitectus carolini (Cystidicolidae), were examined. Camallanus oxycephalus was the only helminth that showed a distinct seasonal pattern. Abundance and prevalence peaked in summer, which was likely driven by concerted reproductive cycles of females. The survey data indicated that the other helminths fluctuated over time but did not seem to follow a distinct seasonal pattern. The fluctuations could be attributed to the variable nature of the habitat or to the stochastic events that influence transmission dynamics, particularly isolated events such as tropical storms, which cause dramatic changes in salinity. The live-box experiment successfully measured recruitment of A. tenauicollis and suggested a seasonal component of the distribution of this species, which was possibly caused by temperature-dependent emergence of cercariae from the snail intermediate host. PMID- 12760650 TI - Amazonspora hassar n. gen. and n. sp. (phylum Microsporidia, fam. Glugeidae), a parasite of the Amazonian teleost Hassar orestis (fam. Doradidae). AB - We describe the microsporidian Amazonspora hassar n. gen., n. sp. from the gill xenomas of the teleost Hassar orestis (Doradidae) collected in the estuarine region of the Amazon River. The parasite appeared as a small whitish xenoma located in the gill filaments near the blood vessels. Each xenoma consisted of a single hypertrophic host cell (HHC) in the cytoplasm of which the microsporidian developed and proliferated. The xenoma wall was composed of up to approximately 22 juxtaposed crossed layers of collagen fibers. The plasmalemma of the HHC presented numerous anastomosed, microvilli-like structures projecting outward through the 1-3 first internal layers of the collagen fibrils. The parasite was in direct contact with host cell cytoplasm in all stages of the cycle (merogony and sporogony). Sporogony appears to divide by plasmotomy, giving rise to 4 uninucleate sporoblasts, which develop into uninucleate spores. The ellipsoidal spores measured 2.69 +/- 0.45 x 1.78 +/- 0.18 microm, and the wall measured approximately 75 nm. The anchoring disk of the polar filament was subterminal, being shifted laterally from the anterior pole. The polar filament was arranged into 7-8 coils in a single layer in the posterior half of the spore, surrounding the posterior vacuole. The polaroplast surrounded the uncoiled portion of the polar filament, and it was exclusively lamellar. The spores and different life cycle stages were intermingled within the cytoplasm of the HHC, surrounding the central hypertrophic deeply branched nucleus. The ultrastructural morphology of this microsporidian parasite suggests the erection of a new genus and species. PMID- 12760649 TI - The U5/U6 snRNA genomic repeat of Taenia solium. AB - The U6 and U5 snRNA (small nuclear ribonucleic acid) genes were identified in Taenia solium with the aim of characterizing their sequence and genomic structures. They are contained within a shared 1,009-nt tandem genomic repeat and present at approximately 3 copies per haploid genome. The U6 snRNA gene shares 92 and 95% sequence similarity with the U6 homologs from humans and Schistosoma mansoni, respectively. The U5 snRNA gene of T. solium is 70% similar to the human U5 sequence in the 5' stem and loop 1 domains. The U6 and U5 snRNA genes are on complementary genomic strands and separated by 458 nt at their "heads" and 306 nt at their "tails." The nucleotides upstream of the U6 gene lack a recognizable TATA box and proximal sequence elements (PSEs), and the putative gene promoter for U5 snRNA does not resemble vertebrate examples. There are short blocks of similarity between the sequences upstream of the U5 and U6 snRNA genes, and these may be sites of shared transcription factor binding at the respective RNA polymerase II and III promoters. It is possible that this unusual allied U5/U6 snRNA genomic repeat may help mediate coordinated regulation of expression of the 2 snRNAs. PMID- 12760651 TI - Icosiella turgeocauda n. sp. (Nematoda: Onchocercidae) and Seuratascaris numidica (Nematoda: Ascarididae), parasites of the frog, Rana cancrivora (Anura: Ranidae), from Luzon, Republic of the Philippines. AB - Icosiella turgeocauda n. sp. from the intestinal mesenteries of Rana cancrivora collected at Luzon, Republic of the Philippines, is described and illustrated. Icosiella turgeocauda n. sp. represents the ninth species to be assigned to the genus and is easily differentiated from all the previously described species by the position of the vulva and the presence of bilateral umbos on the caudal end of the male. Seuratascaris numidica also was found. The Philippines represents a new location record for S. numidica. PMID- 12760652 TI - Phyllodistomum funduli n. sp. (Trematoda: Gorgoderidae) from Fundulus sciadicus Cope from Cedar Creek in western Nebraska. AB - Phyllodistomum funduli n. sp. is described from the urinary bladder and ureters of the plains topminnow, Fundulus sciadicus, from Cedar Creek in Keith County, Nebraska (41 degrees 11.18'N, 101 degrees 21.77'W). Phyllodistomum funduli differs from most other Phyllodistomum species reported in possessing an oral sucker that is larger than the acetabulum. Currently, 7 other species of Phyllodistomum have been reported to possess this trait. A comparison of P. funduli with these 7 other species indicates that P. funduli is a distinct species, lacking notches, posterior body folds, caudal projections, or cephalic glands found in some congeners. There is no published report on parasites other than monogeneans inhabiting the plains topminnow, and there is no other report of members from the Gorgoderidae inhabiting any members of the Fundulidae. PMID- 12760653 TI - A new species of Spauligodon (Nematoda: Pharyngodonidae) parasite of Cnemidophorus spp. (Lacertilia: Teiidae) from southern Mexico. AB - A new species of Spauligodon collected from Cnemidophorus mexicanus and C. deppii is described in this study. The species is placed in Spauligodon because the caudal alae start at the level of the precloacal papillae and embed the adcloacal papillae. The new species is most similar to S. goldbergi, but diagnostic traits of the former are the presence of 2 ridges in lateral alae, flaplike expansions in the caudal end of lateral alae, and a smooth tail for males. This species is the eighth recorded in the neotropics. PMID- 12760654 TI - Rhabdochona ahuehuellensis n. sp. (Nematoda: Rhabdochonidae) from the Balsas goodeid, Ilyodon whitei (Osteichthyes: Gooedeidae), in Mexico. AB - Rhabdochona ahuehuellensis n. sp. is described from the Balsas goodeid, Ilyodon whitei. The distinctive characters are cuticular elevations in the middle of the prostom, tricuspid ending of the distal part of the left spicule combined with a scoop shape, and an egg surface with limited, short subpolar filaments. The characters place the present species farther apart from species described from North America, except R. lichtenfelsi. This description brings the total number of species of Rhabdochona in the Mesa Central of Mexico to 4, suggesting that this genus has undergone speciation in response to basin fragmentation within the Mesa Central of Mexico. PMID- 12760655 TI - Rediscovery and redescription of Plasmodium pifanoi and description of two additional Plasmodium parasites of Venezuelan lizards. AB - Plasmodium pifanoi Scorza and Dagert B., known only from the type host, Ameiva ameiva, is redescribed from Kentropyx calcarata collected in Territorio Amazonas, Venezuela. Schizonts, 6.2 x 4.5 (4-8 x 3-6), produce on average 11.9 (7-16) merozoites. Gametocytes average 12.4 x 6.0 (8-16 x 4-10), with length x width (LW) 72.9 (52-112) and L/W 2.18 (1.1-3.3), and always contain 1-5 prominent vacuoles. Macrogametocytes in active infection are longer than microgametocytes, with greater LW, but gametocytes in chronic infection are not sexually dimorphic in dimension and are slightly smaller. Two additional malarial parasites are described from K. calcarata. Plasmodium lepidoptiformis has small schizonts, 4.6 x 3.2 (3-6 x 2.5-3), that produce 5.1 (4-8) merozoites and commonly resemble a butterfly in appearance. Gametocytes are elongate, 9.0 x 4.3 (7-10 x 3-6), with LW 38.3 (24-51) and L/W 2.2 (1.3-3.3), and sexually dimorphic, with macrogametocytes longer than microgametocytes, with greater LW. Plasmodium minasense calcaratae is characterized by very small, usually fan-shaped, schizonts. 3.4 x 2.6 (2.5-4.5 x 2.0-3.0), that produce 3.9 (3-4) merozoites. Gametocytes are spherical or ovoid, 6.7 x 5.0 (4.5-9.0 x 3.0-7.0), with LW 33.7 (15-54) and L/W 1.4 (1.0-2.3), with no sexual dimorphism in dimensions. PMID- 12760656 TI - Evaluation of analyzing methods for Giardia and Cryptosporidium in a Taiwan water treatment plant. AB - Giardia sp. and Cryptosporidium sp. have emerged as waterborne pathogens of concern in Taiwan. This study examined both parasites in the actual water samples in southern Taiwan. Method 1623 was characterized by a higher recovery rate and lower detection limit compared with the information collection requirement protozoan method. A significant correlation between water turbidity and Cryptosporidium sp. in raw water samples was found in this study. PMID- 12760657 TI - Cloning and characterization of the Leishmania (Viannia) braziliensis Hsp70 gene. Diagnostic use of the C-terminal fragment rLb70(513-663). AB - Cross-reactions between Leishmania braziliensis and Trypanosoma cruzi caused by common antigenic determinants hinder the specific diagnosis of cutaneous and mucocutaneous leishmaniasis (CL and MCL). Therefore, the usefulness of the 70-kDa heat shock protein (Hsp70) from L. braziliensis for differential serodiagnosis was investigated. The single-copy gene encoding Hsp70, consisting of 663 amino acids, was isolated from a genomic DNA library. The antigenicity data show that Hsp70 is an immunodominant antigen highly recognized (84%) by sera of patients with CL and MCL and to a lesser extent by chagasic patients (18.75%). Antigenic mapping of the 5 overlapping fragments into which the protein was split showed that the main antigenic determinants are located in the carboxy-terminal end. The linear antigenic determinants that show cross-reactions with chagasic sera are located in the fragment rLb70(352-518). The carboxy-terminal fragment rLb70(513 663) presents 70% sensitivity and 100% specificity, so it could be a potential candidate for specific serodiagnosis of CL and MCL caused by L. braziliensis. PMID- 12760658 TI - Monozoic cysts of Hepatozoon canis. AB - Small monozoic cysts found in the spleen of dogs infected with Hepatozoon canis are described from naturally and experimentally infected dogs. These forms of H. canis resemble cysts formed by other Hepatozoon species that infect frogs, lizards, and grey squirrels as intermediate hosts. The H. canis cyst stage differs in size and morphology from the large cysts of H. americanum, the second Hepatozoon species known to infect dogs. PMID- 12760659 TI - Upregulation of cardiac cell plasma membrane calcium pump in a canine model of Chagas disease. AB - We have previously demonstrated that cardiac myocytes isolated from the hearts of adult dogs develop rapid repetitive cytosolic Ca2+ transients, membrane depolarization, and cell contraction by mobilization of sarcoplasmic reticulum Ca2+ stores when exposed to a soluble factor from the trypomastigotes of Trypanosoma cruzi. These findings led us to investigate the regulatory mechanisms of cytosolic Ca2+ in cardiac tissues from dogs chronically infected with T. cruzi. Expression of the plasma membrane calcium pump (PMCA) RNA and protein was determined by Northern and Western blotting, respectively, followed by densitometric analyses. A 642-bp PMCA 1b complementary DNA probe derived from canine epicardial tissue hybridized to 2 major transcripts (7.3 and 5.3 kb) in canine epicardium. Expression of the dominant transcript (7.3 kb) was 77% greater in cardiac tissues obtained from dogs with chronic T. cruzi infection (140 days after inoculation) in comparison with constitutive expression levels in normal dogs. Monoclonal antibody 5F10, known to recognize all isoforms of the PMCA, was used to detect expression of the PMCA protein in epicardial tissue. Expression of a 142-kDa protein was increased by 58% in the cardiac tissues of infected dogs when compared with those from uninfected dogs. To establish a link between the upregulation of PMCA in dogs chronically infected with Chagas disease and the ventricular-based arrhythmias and myocardial failure that occur during this stage of disease both in dogs and humans, further study will be required. PMID- 12760661 TI - Temporal and spatial variation of hematozoans in Scandinavian willow warblers. AB - We examined temporal and geographical distribution of Haemoproteus sp. and Plasmodium sp. parasites in Swedish willow warblers, Phylloscopus trochilus. Parasite lineages were detected with molecular methods in 556 birds from 41 sites distributed at distances up to 1,500 km. Two mitochondrial lineages of Haemoproteus sp. were detected, WW1 in 56 birds and WW2 in 75 birds, that differed by 5.2% sequence divergence. We discuss the reasons behind the observed pattern of variation and identify 3 possible causes: (1) variation in the geographic distribution of the vector species, (2) the degree of parasite sharing with other bird species coexisting with the willow warbler, and (3) timing of transmission. Our results support a fundamental and rarely tested assumption of the now classical Hamilton-Zuk hypothesis of sexual selection, namely, that these parasites vary in both time and space. Such fluctuations of parasites and the selection pressure they supposedly impose on the host population are likely to maintain variation in immune system genes in the host population. PMID- 12760660 TI - Characterization of a serine protease activity in Sarcocystis neurona merozoites. AB - Sarcocystis neurona merozoites were examined for their ability to invade and divide in bovine turbinate (BT) cell cultures after treatment with cysteine (iodoacetamide), aspartic (pepstatin A), metallo-(1,10-phenanthroline and ethylene glycol-bis(aminoethylether)-tetraacetic acid [EGTA]), or serine (4-[2 aminoethyl]-benzenesulfonyl fluoride hydrochloride [AEBSF], phenylmethane sulphonyl fluoride [PMSF], and tosyl lysyl chloramethyl ketone [TLCK]) protease inhibitors. Significant (P < 0.01) inhibition of serine protease activity by PMSF and TLCK led to a reduction of 86 and 78% in merozoites produced in BT cell cultures, respectively, whereas AEBSF (1 mM) led to a 68% reduction in merozoites produced in BT cell cultures and a reduction of 84 and 92% at higher AEBSF concentrations (2 and 3 mM, respectively). Pepstatin A and iodoacetamide failed to cause any inhibition in merozoite production, whereas 1,10-phenanthroline and EGTA caused slight, but not significant, inhibition at 6 and 17%, respectively. In zymograms, 2 bands of protease activity between 65- and 70-kDa molecular weight were seen. The protease activity was inhibited by AEBSF but not by E-64 (cysteine protease inhibitor), EGTA, iodoacetamide, or pepstatin A. In native zymograms, the protease activity was highest between a pH range of 8 and 10. These data suggest that merozoites of S. neurona have serine protease activity with a relative molecular weight range between 65 and 70 kDa and optimal pH range between 8 and 10, which is essential for host cell entry at least in vitro. The protease activity described here could be a potential target for chemotherapy development. PMID- 12760662 TI - Cryptosporidium parvum infection in gene-targeted B cell-deficient mice. AB - The importance of B cells in host resistance to, and recovery from, Cryptosporidium parvum infection was examined in gene-targeted B cell-deficient (muMT-/-) mice. Neonatal muMT-/- mice infected with C. parvum at 5 days of age completely cleared the infection by day 20 PI. The kinetics of infection and clearance were similar to those seen with age-matched C57BL/6 control mice. Furthermore, B cells were not required to clear existing C. parvum infection in adult mice. Reconstitution of persistently infected Rag-1-/- adult mice with spleen cells from muMT-/- donor mice resulted in significant reduction of infection, as in the results seen with spleen cells from C57BL6 donors. These findings indicate clearly that B cells are not essential for host resistance to, and recovery from, C. parvum infection in mice. PMID- 12760663 TI - Experimental Sarcocystis hominis infection in a water buffalo (Bubalus bubalis). AB - A water buffalo (Bubalus bubalis) was fed 5.0 x 10(5) Sarcocystis hominis sporocysts from a human volunteer who had ingested S. hominis cysts from naturally infected cattle. A necropsy was performed on the buffalo 119 days after inoculation, and a large number of microscopic sarcocysts (approximately 5,000/g) were found in skeletal muscles. Ultrastructurally, the sarcocyst wall from buffalo muscles has upright villar protrusions measuring about 5.6 x 0.8 microm with numerous microtubules that run from the base to the apex. Sarcocysts from this buffalo were infective to 2 human volunteers, confirming their identity as S. hominis. Therefore, we believe that buffaloes can act experimentally as the intermediate host for S. hominis. PMID- 12760664 TI - Prevalence of Toxoplasma gondii in chickens from an area in southern Brazil highly endemic to humans. AB - The prevalence of Toxoplasma gondii in free-range chickens from Campos dos Goytacazes, Rio de Janeiro State, Brazil, was examined to evaluate environmental contamination by oocysts. Antibodies against T. gondii were assayed by the modified agglutination test (MAT) in sera of chickens. Antibodies against the parasite were found in 129 of 198 chickens with MAT titers > or = 1:25. Brains and hearts of 86 of the 198 chickens were bioassayed in mice for the presence of T. gondii. Viable parasites were isolated from 61 (70.9%) of the 86 chickens. Importantly, viable T. gondii were recovered even from seronegative chickens (MAT titer < or = 1:10). The distribution of parasite-positive chickens by MAT titer was 4 of 17 (titer < or = 1:10), 3 of 4 (titer of 1:20), 2 of 6 (titer of 1:40), and 52 of 59 (titer > or = 1:80). Thus, the high recovery rate of T. gondii observed in mice is indicative of high levels of environmental contamination of free-range chickens by T. gondii oocysts in this area that is endemic to humans. PMID- 12760665 TI - Sarcocysts of an unidentified species of Sarcocystis in the sea otter (Enhydra lutris). AB - The number of Sarcocystis species that infect sea otters (Enhydra lutris) is unknown. Sea otter tissues were recently shown to harbor sarcocysts of S. neurona and of unidentified species of Sarcocystis. Whereas sarcocysts of S. neurona have walls 1-3 microm thick with type 9 villar protrusions, ultrastructure of a distinct thin-walled sarcocyst (0.5-0.7 microm thick) lacking villar protrusions, but instead exhibiting minute type 1 undulations on the sarcocyst wall, is described in this report. Parasites characterized from a sea otter infection were inferred to be related to, but distinct from, other species belonging to Sarcocystis, based on sequencing and phylogenetic analysis of a portion of the beta subunit of the plastid-encoded RNA polymerase gene. PMID- 12760666 TI - Morphology is not a reliable tool for delineating species within Cryptosporidium. AB - Within the coccidia, morphological features of the oocyst stage at the light microscope level have been used more than any other single characteristic to designate genus and species. The aim of this study was to conduct morphometric analysis on a range of Cryptosporidium spp. isolates and to compare morphological data between several genotypes of C. parvum and a second species C. canis, as well as a variation within a specific genotype (the human genotype), with genetic data at 2 unlinked loci (18S ribonucleic deoxyribonucleic acid and HSP 70) to evaluate the usefulness of morphometric data in delineating species within Cryptosporidium. Results indicate that morphology could not differentiate between oocysts from C. parvum genotypes and oocysts from C. canis, whereas genetic analysis clearly differentiated between the two. The small size of the Cryptosporidium spp. oocyst, combined with the very limited characters for analysis, suggests that more reliance should be placed on genetic differences, combined with biological variation, when delineating species within Cryptosporidium. PMID- 12760667 TI - Molecular cloning of a novel multidomain Kunitz-type proteinase inhibitor from the hookworm Ancylostoma caninum. AB - Degenerate oligonucleotide primers derived from conserved serine protease inhibitors were used to amplify a 90-base pair (bp) amplicon from an Ancylostoma caninum adult-stage complementary deoxyribonucleic acid (cDNA) library by polymerase chain reaction (PCR). The amplicon was labeled and used as a probe to screen the library, and a 2,300-bp cDNA clone was identified. The 5' end of the molecule was obtained from adult cDNA by 5'-RACE. The complete sequence named A. caninum Kunitz-type protease inhibitor (Ac-kpi-1) was 2,371 bp and encoded a 759 amino acid open reading frame. The deduced amino acid sequence had a calculated molecular weight of 84,886 Da and contained an amino terminal signal peptide, suggesting that the protein is secreted. Analysis of the predicted protein sequence indicates 12 highly conserved Kunitz-type serine protease inhibitor domains connected by short, conserved spacers. On the basis of sequence analysis, the first 11 domains are predicted to be active serine protease inhibitors based on the P1 amino acid. Domains 5-8 have identical amino acid sequences, and the remaining domains are 38-88% identical. Domain 12 lacks several of the conserved cysteine residues and has an atypical amino acid in the P1 position, suggesting that it is nonfunctional. Reverse transcriptase-PCR indicated that the Ac-kpi-1 messenger ribonucleic acid is present in egg, L1, L3, and adult stages but is most abundant in the adult stage. Ac-KPI-1 is most similar in domain architecture to several extracellular matrix proteins involved in cellular remodeling during insect development. In addition, there are 44 nematode proteins containing one or more Kunitz domains in GenBank, including several with multiple domains. PMID- 12760668 TI - A human case of gnathostomiasis nipponica confirmed indirectly by finding infective larvae in leftover largemouth bass meat. AB - A human case of creeping eruption due to Gnathostoma nipponicum was confirmed indirectly by finding infective advanced third-stage larvae in leftover largemouth bass meat. This is the first report indicating that the largemouth bass (Micropterus salmoides) serves as a source of G. nipponicum infection in humans. PMID- 12760669 TI - Two species of canine Babesia in Australia: detection and characterization by PCR. AB - The haemoprotozoan Babesia canis has been recognized in Australia for many years, and a second, smaller species has recently been discovered. Amplification and sequencing of a partial region of the 18S small subunit ribosomal RNA (rRNA) gene enabled detection and characterization of the large and small canine babesiae of Australia for the first time. Isolates from northern Australia were genetically characterized to be 99% homologous to Babesia canis vogeli, confirming previous speculation about the subspecies of B. canis endemic to Australia. The partial 18S rRNA gene sequence amplified from isolates obtained in southeastern Australia was genetically identical to Babesia gibsoni, a species not previously known in Australia. The polymerase chain reaction (PCR) used was shown to be specific to Babesia and had a high sensitivity, detecting DNA at a parasitemia of approximately 0.0000027%. This study also reports the first known detection and characterization of B. canis DNA in Rhipicephalus sanguineus ticks using PCR. PMID- 12760670 TI - Cutaneous trematode Collyriclum faba in wild birds in the central European Carpathians. AB - The occurrence of cutaneous trematode Collyriclum faba in wild birds was monitored in the central European Carpathians from 1996 to 2001. A total of 5,414 birds, representing 86 species, was examined. Collyriclum faba was found at 7 sites (5 in Slovakia, 1 in Poland, and 1 in the Czech Republic), and prevalences at the sites varied from 1 to 16%. Ten species of passerine birds were infected: blackcap (Sylvia atricapilla) (16 positive/622 tested, 2% prevalence), black redstart (Phoenicurus ochruros) (2/25, 8%), chaffinch (Fringilla coelebs) (7/113, 6%), common blackbird (Turdus merula) (1/143, 1%), common redstart (Phoenicurus phoenicurus) (1/30, 3%), dipper (Cinclus cinclus) (1/9, 11%), European robin (Erithacus rubecula) (103/838, 12%), goldcrest (Regulus regulus) (1/76, 1%), grey wagtail (Motacilla cinerea) (5/25, 20%), and yellowhammer (Emberiza citrinella) (1/73, 1%). Cutaneous cysts of C. faba were found in the birds from the end of May to mid-September, with the prevalence peaking in July and August. One to 21 cysts per bird were found. In black redstart, chaffinch, common redstart, European robin. and yellowhammer, cysts were most frequently observed on the legs, particularly in the crural region. In blackcap, common blackbird, dipper, and grey wagtail, almost all the cysts were found around the vent and on the abdomen. In goldcrest, the cyst was located above the coccygeal gland. Collyriclum faba is a common parasite in birds during summer in central European Carpathians. Collyriclum faba was observed for the first time in Poland. It appears that C. faba may be fatal for some of the bird hosts. PMID- 12760671 TI - First report of a natural hybrid between Schistosoma mansoni and S. rodhaini. AB - Experimental crosses between Schistosoma mansoni and S. rodhaini have shown that hybrid offspring are viable, yet, until now, no naturally occurring hybrid has been identified. A collection of freshwater snails from Nyamlebi-Ngoma, Ukerewe Island, Lake Victoria, Tanzania, yielded a mixed infection within a single Biomphalaria sudanica of S. mansoni females and S. mansoni-S. rodhaini hybrid males. The hybrids were identified using deoxyribonucleic acid (DNA) sequences. Mitochondrial DNA 16S and 12S sequences of the hybrids match those of S. mansoni, whereas their nuclear ribosomal DNA ITS1 and ITS2 sequences match those of S. rodhaini. The identification of hybrids in Tanzania highlights the possibility that the genetic identity of either parasite species might be modified by introgression. PMID- 12760673 TI - Field method for isolation of trichostrongyle larvae from vegetation of natural pastures of Arctic ruminants. AB - The extent to which wild ruminant populations are exposed to infective helminth larvae on their natural pastures is relatively undetermined. In the present study, a modified method for sampling of herbage and isolation of trichostrongyle infective third-stage larvae from natural pastures was used successfully in a muskox habitat in low-Arctic Greenland. The method, a revision of the Macro Baermann method, is particularly aimed at fieldwork under primitive conditions. PMID- 12760672 TI - Cellular immune responses in chickens induced by recombinant R7 Leucocytozoon caulleryi vaccine. AB - We have recently developed recombinant subunit vaccine consisting of second generation schizont (2GS) membrane protein (rR7) of Leucocytozoon caulleryi. Chickens immunized with rR7 antigen acquired clear resistance to challenge by Leucocytozoon sporozoites. We examined the induction of cellular immune responses in vaccinated chickens. Spleen adherent cells from vaccinated chickens showed significantly higher phagocytic activity against 2GS-coated latex particles than did cells from adjuvant-inoculated or untreated control birds. Anti-R7 chicken IgG significantly increased the phagocytic rate of adherent cells from these 3 groups. These results show that specific cellular immune responses are induced by recombinant R7 subunit vaccine consisting of L. caulleryi 2GS protein, which suppresses the growth of parasites in the host in association with antiparasite antibodies to 2GS antigen. PMID- 12760674 TI - Polymerase chain reaction for the detection of Cryptosporidium spp. in cat feces. AB - The objective of this study was to compare a polymerase chain reaction (PCR) assay and a monoclonal antibody-based immunofluorescence assay (IFA) for detection of Cryptosporidium parvum in cat feces. Eight C. parvum-naive DSH cats were orally inoculated with 1 x 10(6) oocysts of a C. parvum human isolate. Fecal samples were collected before inoculation, daily for the next 30 days, and twice weekly until day 85. Methylprednisolone acetate was administered at 20 mg/kg i.m. on days 85, 92, and 99. From days 86 to 115, feces were collected daily and then up to twice weekly until day 126. Immunofluorescence assay was performed after collection of the samples, and then the samples were frozen at -70 C until assayed by PCR. Cryptosporidium parvum was detected by PCR in 101 of 353 samples and by IFA in 52 of 353 samples: 27 samples were PCR positive, IFA positive; 74 samples were PCR positive, IFA negative; 25 samples were PCR negative, IFA positive; and 227 samples were PCR negative, IFA negative. The percentage of concordance between IFA and PCR was 72%. Results of this study suggest that this PCR assay is more sensitive than IFA for detection of C. parvum in cat feces. PMID- 12760675 TI - ICM0201, a new inhibitor of osteoclastogenesis from Cunninghamella sp. F-1490. I. Taxonomy, fermentation, isolation and biological activities. AB - In the course of screening for inhibitors of osteoclastogenesis, a new substance designated as ICM0201 was isolated from a fermentation broth of Cunninghamella sp. F-1490. ICM0201 inhibited the formation of osteoclasts in mouse bone marrow cells with an IC50 value of 0.78 microg/ml and showed weak cytotoxicity against bone marrow cells. PMID- 12760676 TI - ICM0201, a new inhibitor of osteoclastogenesis from Cunninghamella sp. F-1490. II. Structure determination and synthesis. AB - ICM0201 (1), a new inhibitor of murine osteoclastogenesis in culture was isolated from a fermentation broth of Cunninghamella sp. F-1490. The structure of ICM0201 was determined to be (3S,10aR)-3,4a-dihydroxy-2,3,4,4a-tetrahydro-2H-pyrano[3,2 b]benzo[e]morpholine-9-carboxylic acid by spectroscopic analyses and chemical studies. The structure of 1 is unique in that the tricycle ring system is composed of aminal and hemiacetal bonds. PMID- 12760677 TI - IB-00208, a new cytotoxic polycyclic xanthone produced by a marine-derived Actinomadura. I. Isolation of the strain, taxonomy and biological activites. AB - A new compound, IB-00208, has been isolated from the fermentation broth of an actinomycete isolated from a marine environment. The strain was identified as Actinomadura sp. by its chemical and phylogenetic characteristics. The compound shows cytotoxic activity on tumor cell lines and bactericidal activity against gram-positive bacteria. PMID- 12760678 TI - Nocathiacins, new thiazolyl peptide antibiotics from Nocardia sp. I. Taxonomy, fermentation and biological activities. AB - Thiazolyl peptide antibiotics, nocathiacin I, II and III, were identified in a culture of Nocardia sp. WW-12651 (ATCC 202099). They exhibit potent in vitro activity (ng/ml) against a wide spectrum of gram-positive bacteria, including multiple-drug resistant pathogens such as methicillin-resistant Staphylococcus aureus (MRSA), multi-drug resistant Enterococcus faecium (MREF) and fully penicillin-resistant Streptococcus pneumoniae (PRSP), and demonstrate excellent in vivo efficacy in a systemic Staphylococcus aureus infection mice model. PMID- 12760679 TI - Nocathiacins, new thiazolyl peptide antibiotics from Nocardia sp. II. Isolation, characterization, and structure determination. AB - A new group of thiazolyl peptide antibiotics, the nocathiacins, was isolated from cultured broth of Nocardia sp. The major analogs nocathiacins I-III (1-3) were purified using silica gel and Sephadex LH-20 chromatography techniques. The structures of nocathiacins I-III were determined by spectroscopic (2D-NMR, MSn) methods, and share structural similarities to glycothiohexide-alpha (4). PMID- 12760681 TI - Studies on novel bacterial translocase I inhibitors, A-500359s. II. Biological activities of A-500359 A, C, D and G. AB - A-500359 A, C, D, G and capuramycin inhibited bacterial phospho-N-acetylmuramyl pentapeptide-translocase (translocase I: EC 2.7.8.13) with IC50 values of 0.017, 0.12, 0.53, 0.14 and 0.018 microM, respectively. Consistently, A-500359 A, C and capuramycin inhibited in vitro peptidoglycan biosynthesis. A-500359 A exhibited reversible inhibition, which was mixed type and noncompetitive with respect to UDP-MurNAc-(N(epsilon)-Dns)pentapeptide (Ki=0.0079 microM) and undecaprenyl phosphate, respectively. A-500359 A, C, D and G showed antimicrobial activity against Mycobacterium smegmatis. As a single intravenous injection of A-500359 A at a dose of 500 mg/kg showed no toxicity in mice, it was suggested that the capuramycin derivatives might become candidates as novel therapeutic agents for various diseases caused by Mycobacteria including tuberculosis. PMID- 12760680 TI - Studies on novel bacterial translocase I inhibitors, A-500359s. I. Taxonomy, fermentation, isolation, physico-chemical properties and structure elucidation of A-500359 A, C, D and G. AB - In the course of our screening for bacterial phospho-N-acetylmuramyl-pentapeptide translocase (translocase I: EC 2.7.8.13) inhibitors, we found inhibitory activity in the cultured broth of the strain identified as Streptomyces griseus SANK 60196. The strain produced capuramycin and four novel capuramycin derivatives designated as A-500359 A, C, D and G. Purification and structural analysis were performed, and the structures of A-500359 A, C, D and G were elucidated as 6''' methylcapuramycin, 3'-demethyl-6'''-methylcapuramycin, 2''-deoxy-6''' methylcapuramycin and 3'-demethylcapuramycin, respectively. PMID- 12760682 TI - Studies on novel bacterial translocase I inhibitors, A-500359s. III. Deaminocaprolactam derivatives of capuramycin: A-500359 E, F, H; M-1 and M-2. AB - Novel derivatives of capuramycin were obtained when 10 mM of 2-aminoethyl-L cysteine (AEC), an inhibitor of aspartokinase, was added to the culture of Streptomyces griseus SANK 60196, the producer of A-500359. They were purified from the culture filtrate and their chemical structures were elucidated as a deaminocaprolactam derivative of capuramycin designated as A-500359 F, A-500359 E, a methyl ester of A-500359 F, and A-500359 H, a 3'-demethyl derivative of A 500359 F. Two other compounds, A-500359 M-1 and A-500359 M-2, were purified from the same medium and their structures were elucidated. A-500359 E, F, H, M-1 and M 2 inhibited bacterial translocase I with an IC50 of 0.027 microM, 1.1 microM, 0.008 microM, 0.058 microM and 0.010 microM, respectively. A-500359 E, M-1 and M 2 inhibited the growth of mycobacteria as well. PMID- 12760683 TI - Studies on novel bacterial translocase I inhibitors, A-500359s. IV. Biosynthesis of A-500359s. AB - This report describes the isolation of novel A-500359 analogues from the culture broth of Streptomyces griseus SANK 60196 and 13C-incorporation studies of A 500359 A to reveal the biosynthetic pathway of A-500359 derivatives. As a result, A-500359 M-3 and J were isolated as novel analogues. The former, isolated from a culture broth fed with unnatural amino acids, was a novel amino acid adduct of A 500359, and the latter was found to be a putative precursor of all A-500359 derivatives, on the basis of the structure. Moreover, 13C-incorporation studies revealed the origin of every carbon atom of A-500359 A. From these results, it was revealed that the core skeleton of A-500359 was biosynthesized from uridine and phosphoenolpyruvate in the same manner as for polyoxin, a nucleoside antibiotic. Moreover, the uronic acid and aminocaprolactam moiety was derived from hexose and lysine, respectively, and two methyl groups of A-500359 A were derived from methionine. PMID- 12760684 TI - Isolation and structure of a new macrolide antibiotic, erythromycin G, and a related biosynthetic intermediate from a culture of Saccharopolyspora erythraea. AB - The new naturally occurring erythromycin G (4), formally derived from erythromycin B by hydroxylation of the C-16 methyl group, and 3-O mycarosylerythronolide B (5), an erythromycin biosynthetic intermediate previously obtained only from microorganisms blocked in erythromycin biosynthesis, were isolated from a concentrate of mother liquors derived from a culture of Saccharopolyspora erythraea. The structure of erythromycin G was defined by spectroscopic data and X-ray crystallographic analysis. Theoretical calculation of 4 has been performed at MM2 level, and the low-energy conformations have been compared with X-ray data: both theoretical and experimental approaches give similar three-dimensional shapes. Antibacterial activity of 4 against both gram-positive and gram-negative organisms has been evaluated. A simple method for the isolation of large amounts of erythromycins B (2) and D is provided as well. PMID- 12760685 TI - Glycopeptide carboxamides active against vancomycin-resistant enterococci. AB - Glycopeptide antibiotics were synthesized via the PyBOP mediated condensation of aliphatic, heterocyclic and aromatic amines with the C-terminus of vancomycin, LY264826 (A82846B) and semi-synthetic derivatives of these natural products. Amides of LY264826 and vancomycin demonstrated excellent activity against staphylococci and streptococci as compared to the parent natural product. However, the amides of N-alkylated LY264826 and N-alkylated vancomycin were active against vancomycin-resistant enterococci as well as other gram-positive pathogens such as Staphylococcus aureus, S. haemolyticus, S. epidermidis and Streptococcus pneumoniae. PMID- 12760686 TI - Synthesis and structure-activity relationship of antifungal coniothyriomycin analogues. AB - The structure of the antifungal metabolite coniothyriomycin was systematically modified by changing the acids of the open chain imide, modification of the hydrophobicity, variation in the degree of saturation, replacement of carbons by nitrogen or oxygen, and incorporation of the open chain molecule into cyclic arrangements. Structure-activity studies showed that antifungal activity was retained by replacement of phenylacetic acids by benzoic acids in the imide structure but diminished by hydrogenation of the fumaric ester part. PMID- 12760687 TI - 1100-50, a novel nematocide from Streptomyces lavendulae SANK 64297. PMID- 12760688 TI - Production of demannosyl-A40926 by a Nonomuraea sp. ATCC 39727 mutant strain. PMID- 12760690 TI - IB-00208, a new cytotoxic polycyclic xanthone produced by a marine-derived Actinomadura. II. Isolation, physico-chemical properties and structure determination. PMID- 12760689 TI - UK-2A, B, C, and D, novel antifungal antibiotics from Streptomyces sp. 517-02. VIII. Reactive oxygen species generated by C9-UK-2A, a derivative of UK-2A, in Rhodotorula mucilaginosa IFO 0001. PMID- 12760692 TI - The second mouse. PMID- 12760691 TI - In vitro antimalarial activities of the microbial metabolites. PMID- 12760693 TI - Buonocore memorial lecture. Adhesion to enamel and dentin: current status and future challenges. AB - Bonding to tooth tissue can be achieved through an "etch&rinse," "self-etch" or "glass-ionomer" approach. In this paper, the basic bonding mechanism to enamel and dentin of these three approaches is demonstrated by means of ultramorphological and chemical characterization of tooth-biomaterial interfacial interactions. Furthermore, bond-strength testing and measurement of marginal sealing effectiveness (the two most commonly employed methodologies to determine "bonding effectiveness" in the laboratory) are evaluated upon their value and relevance in predicting clinical performance. A new dynamic methodology to test biomaterial-tooth bonds in a fatigue mode is introduced with a recently developed micro-rotary fatigue-testing device. Eventually, today's adhesives will be critically weighted upon their performance in diverse laboratory studies and clinical trials. Special attention has been given to the benefits/drawbacks of an etch&rinse versus a self-etch approach and the long-term performance of these adhesives. Correlating data gathered in the laboratory with clinical results clearly showed that laboratory research CAN predict clinical effectiveness. Although there is a tendency to simplify bonding procedures, the data presented confirm that conventional three-step etch&rinse adhesives still perform most favorably and are most reliable in the long-term. Nevertheless, a self-etch approach may have the best future perspective. Clinically, when adhesives no longer require an "etch&rinse" step, the application time, and probably more importantly, the technique-sensitivity are substantially reduced. Especially "mild," two-step self-etch adhesives that bond through a combined micromechanical and chemical interaction with tooth tissue closely approach conventional three step systems in bonding performance. PMID- 12760694 TI - In vivo kinetics of bleaching gel with three-percent hydrogen peroxide within the first hour. AB - This in vivo study determined the kinetics of 3% hydrogen peroxide in a bleaching gel within the first hour. The material used in this study was 3% hydrogen peroxide gel (Perfecta 3/15, Premier Dental Products Co) and the study involved 10 subjects who met the inclusion and exclusion criteria. Each subject wore the tray with gel six different times on separate days. Evaluation of the remaining amount of hydrogen peroxide was calculated by the method stated in US Pharmacopoeia. The study results indicate that the mean percentage of hydrogen peroxide recovered for 5, 10, 20, 30, 45 and 60 minutes was 61, 56, 49, 44, 38 and 32, respectively. The amount of hydrogen peroxide in the saliva sample after one hour was 0.42 mg. Excluding the first 10 minutes, the kinetics of hydrogen peroxide in the tray and teeth sample was exponential. PMID- 12760695 TI - Comparison of pulp responses to resin composites. AB - Diverse reports have described how various types of adhesive systems cause disastrous pulp necrosis, chronic severe inflammation or failure to stimulate any pulp reactions. This article reports on the effects of five common adhesive systems and how they compare in terms of pulp injury as measured by odontoblast survival or dentin regeneration and reactionary dentin formation. One hundred and thirty Class V pulp, non-exposed cavities were prepared in non-human primate teeth and were restored with five different adhesive systems. After a period of time between 3 and 172 days, the teeth were extracted, fixed, processed and examined histomorphometrically. Bacterial microleakage was detected with McKays stain and inflammation was categorized according to the International Organization for Standardization (ISO) criteria. The number of odontoblasts and the area of reactionary dentin were measured. Pulp reactions of all adhesive systems were generally minimal, although some systems permitted bacterial microleakage in 33% of restorations, and some other systems were associated with pulp inflammation in 22% of restorations. These observations suggest that adhesive systems provide acceptable biocompatibility, however, there is strong potential for improvement. PMID- 12760697 TI - Thermal emission by different light-curing units. AB - This study quantified and compared the thermal emission of different light curing units (LCU). Three LED (Elipar Freelight [3M]; GC e-light [GC]; Coolblu [Dentalsystems.com]) and three halogen (Max [Dentsply-Caulk]; Elipar Trilight [3M]; Astralis 10 [Ivoclar-Vivadent]) lights were selected for the study. Thermal emission of the LCUs, when used in various curing modes, was assessed using a K type thermocouple and a digital thermometer at distances of 3 mm and 6 mm compared to the conventional halogen LCU (Max). The temperature profiles and mean maximum temperature change (n = 7) generated by each LCU were obtained. Data was subjected to ANOVA/Scheffe's post-hoc test and Independent Samples t-test at significance level 0.05. At 3 mm, temperature rise observed with LED lights ranged from 4.1 degrees C to 12.9 degrees C, while halogen lights ranged from 17.4 degrees C to 46.4 degrees C. At 6 mm, temperature rise ranged from 2.4 degrees C to 7.5 degrees C and 12.7 degrees C to 25.5 degrees C for LED and halogen lights, respectively. Thermal emission of LED lights was significantly lower than halogen lights. Significant differences in temperature rise were observed between different curing modes for the same light and between different LED/halogen lights. PMID- 12760696 TI - Cytotoxicity of conventional and modified glass ionomer cements. AB - Various glass ionomer cements (GICs) and resin-modified GICs are widely used as tooth-colored restorative materials. However, their potential effects on pulp tissues are not fully understood. In this study, the authors compared the toxicity of nine types of GICs on cultured human dental pulp cells. Exposure of pulp cells to GICs for five days led to differential growth inhibition as analyzed by 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. Exposure of pulp cells to ProTec CEM, Fuji II LC, Compoglass and GC Lining cement for five days decreased the cell numbers to 11%, 12%, 19% and 25%, respectively, of the control. Exposure of pulp cells to Fuji IX, GIC FX and Fuji II SC also decreased cell numbers by 62%, 33% and 24%, respectively. By contrast, Hy-Bond and Fuji I showed only mild suppression on the growth of pulp cells, with 12% and 16% decreased cell numbers. Morphologically, marked retraction and rounding of pulp cells were noted following exposure to GC Lining cement; in addition, cell surface blebbing was noted following exposure to Compoglass, Fuji II LC and ProTec CEM. Exposure of the pulp cells to Fuji II SC and Fuji IX, however, led to decreases in the cell density, with no obvious morphological changes. These results indicate that resin-modified GICs, such as Compoglass, Fuji II LC, ProTec CEM and GC Lining cements, are more toxic to pulp cells than conventional GICs. It is not recommended that resin-modified GICs be directly applied onto dental pulp cells. However, additional in vivo studies are needed to evaluate the potential toxicities of these resin-modified GICs during clinical operative procedures. PMID- 12760698 TI - Microleakage evaluation of a flowable polyacid-modified resin composite used as fissure sealant on air-abraded permanent teeth. AB - This study evaluated the microleakage of a flowable polyacid-modified resin composite used as a fissure sealant on air-abraded teeth in comparison to a resin based fluoride sealant. Sixty extracted human third molars were divided into six subgroups (n = 10) under two main groups (fissure sealant materials). The occlusal surfaces of the teeth were air-abraded at 80 psi using 50 microm particles of aluminum oxide for 30 seconds, followed by rinsing and drying. Delton FS+ (Dentsply International) and Dyract Flow (Dentsply DeTrey) were applied to the teeth in subgroups following application of: a) an acid etching and bonding agent, b) bonding agent alone or c) air-abrasion alone, respectively. Acid etching (Delton EZ Etch, Dentsply International) and bonding agent application (Prime & Bond NT, Dentsply DeTrey) were done according to the manufacturers' instructions. The teeth were thermocycled for 500 cycles between 5 degrees C and 55 degrees C with dwell time of 15 seconds. Basic fuchsin (0.5%) staining followed by buccolingual sectioning was performed. Microleakage was evaluated at 20x optical magnification. Kruskal-Wallis test was used to make comparisons among six subgroups. Pairwise comparisons were done with the Mann Whitney U test with the level of significance set as alpha = 0.05. Dyract Flow application as a fissure sealant on air-abraded permanent teeth in combination with acid etching and/or bonding agent provided microleakage results comparable to Delton FS+. Results also showed that the use of air abrasion, alone, resulted in significantly higher microleakage scores. PMID- 12760699 TI - Physiological remineralization of artificially demineralized dentin beneath glass ionomer cements with and without bacterial contamination in vivo. AB - This study evaluated the physiological remineralization of artificially demineralized dentin beneath glass ionomer cements with and without bacterial contamination. The artificially demineralized dentin was produced on 84 monkey teeth using a decalcifying solution. Half the samples were left open to the oral cavity for one week, then, all cavities were restored with two glass-ionomer cements: Fuji IX or Fuji II LC improved (n = 7). The nanohardness of the artificially demineralized dentin at 3, 90 and 360 days was measured using a nanoindentation tester (ENT-1100, Elionix) and compared statistically by two-way ANOVA and Fisher's PLSD test (p < 0.05). Each mineral (Ca, Mg, P, F) within the demineralized dentin was also analyzed using Electron Probe Microanalysis. For the samples, the mean nanohardness of the three-day samples was significantly lower than the 360-day samples (p < 0.05). Although there was no significant difference in the mean nanohardness within all the bacterially-contaminated groups through the experimental periods (p > 0.05), the mean nanohardness of the bacterial-contaminated samples were significantly lower than the non-bacteria contaminated samples (p < 0.05). From the EPMA results, fluoride release from both cements to the bottom of the artificially demineralized dentin was detected within three days. Although Ca density was sparse within this demineralized dentin lesion, for the Fuji IX sample, a high Mg density within this lesion was detected at 360 days. PMID- 12760700 TI - Assessing the surface roughness of a posterior resin composite: effect of surface sealing. AB - This study assessed the in vitro influence of surface sealing on the surface roughness of a posterior resin composite before and after toothbrushing. Thirty specimens (13 mm diameter x 1 mm high) were fabricated from Filtek-P60 resin composite and randomly assigned to three groups (n = 10): a non-sealed control and two groups sealed with one of the tested materials-a surface-penetrating sealant (Protect-it!-PI) and a one bottle adhesive system (Single Bond-SB). The samples were subjected to a surface roughness reading to determine the initial roughness, then submitted to simulated toothbrushing with 35,600 cycles for 100 minutes. Specimens were then cleaned and a post-abrasion surface roughness reading accomplished. Means (microm), recorded before (B) and after (A) toothbrushing, and standard deviations were: Control--(B): 0.032 (+/- 0.005), (A): 0.054 (+/- 0.005); PI-(B): 0.034 (+/- 0.005), (A): 0.060 (+/- 0.034); SB (B): 0.031 (+/- 0.004), (A): 0.047 (+/- 0.007). Data were tabulated and submitted to two-way ANOVA. No statistically significant difference was observed when the control and experimental groups were compared. However, a significant difference (p < 0.05) was found between the measurements performed before and after toothbrushing. Based on these results, it may be concluded that using either a surface penetrating sealant or a one bottle adhesive system did not provide the optimization of superficial integrity. The use of a dentifrice and toothbrush resulted in significant alterations to the surface smoothness of the resin composite. PMID- 12760701 TI - The effect on shear bond strength of rewetting dry dentin with two desensitizers. AB - The difficulty related to achieving a balance between wet and dry dentin makes the dentin bonding technique extremely sensitive. This study evaluated the effect of rewetting dried dentin with two commercial desensitizing agents (Protect and HurriSeal) on the dentin shear bond strength of three total-etch dentin bonding agents (Syntac Single-Component, OptiBond Solo Plus and Prime & Bond NT) and compared both to applying these same bonding agents to moist dentin and dry dentin. Each bonding agent was paired with an appropriate resin composite from the same manufacturer (Table 1). Recently extracted, formalin-treated caries-free human molars were used. The occlusal surface of each tooth was ground to create a flat dentin surface. Then, each tooth was mounted in acrylic. Twelve groups (n = 15) were prepared: 1) Syntac Single-Component with Heliomolar resin composite (SSC/H) to moist dentin; 2) SSC/H to dry dentin; 3) SSC/H to dried dentin rewet with Protect; 4) SSC/H to dried dentin rewet with HurriSeal; 5) OptiBond Solo Plus with Point 4 resin composite (OBS+/P4) to moist dentin; 6) OBS+/P4 to dry dentin; 7) OBS+/P4 to dried dentin rewet with Protect; 8) OBS+/P4 to dried dentin rewet with HurriSeal; 9) Prime & Bond NT with TPH Spectrum resin composite (PBNT/TPH) to moist dentin; 10) PBNT/TPH to dry dentin; 11) PBNT/THP to dried dentin rewet with Protect and 12) PBNT/TPH to dried dentin rewet with HurriSeal. Groups 1, 5 and 9 were placed according to manufacturers' instructions (moist dentin) as control groups. All the other groups received a 15-second air blast after etching and prior to applying the one bottle adhesive or desensitizer and one bottle adhesive. Resin composite cylinders [4 mm in diameter and 2 mm in height] were then placed. The specimens were stored in distilled water at 37 degrees C for 24 hours prior to thermocycling 2,500 times (at 8 degrees C and 48 degrees C). Shear bond strengths (SBSs) were measured one week after fabrication by using a circular knife-edge and crosshead speeds of 0.5 mm/minute. Failure modes of debonded specimens were determined under a stereomicroscope (30x). Failed specimens with the low and high shear bond strengths in each group were evaluated under a low vacuum Scanning Electron Microscope (SEM-LV). One-way ANOVA and Tukey's tests were used to compare the different conditions for each bonding system. In the Syntac Single-Component bonding agent groups, there was no significant difference in shear bond strength between the control (15.73 MPa), dry (18.11 MPa) and HurriSeal (16.18 MPa) specimens. Protect specimens showed significantly lower shear bond strength (6.39 MPa). In the OptiBond Solo Plus bonding agent groups, there was no significant difference between the control (20.79 MPa) and the HurriSeal (21.29 MPa) groups, and both groups had significantly greater bonds than the dry (14.13MPa) and Protect (9.57 MPa) groups. In the Prime & Bond NT bonding agent groups, the shear bond strength of the HurriSeal group (20.73 MPa) was significantly higher than the other groups: control (8.05 MPa), dry (5.73 MPa) and Protect (5.45 MPa). PMID- 12760702 TI - Effect of cooled composite inserts in the sealing ability of resin composite restorations placed at intraoral temperatures: an in vitro study. AB - Polymerization shrinkage causes microleakage of resin composite restorations. New materials and operative techniques should be developed in order to reduce polymerization shrinkage. This research studied the effects of cooled composite inserts and room-temperature composite inserts in the sealing ability of resin composite restorations placed at intraoral temperatures. Forty-eight extracted human molars (providing a total of 155 sections) were kept at intraoral temperatures, and Class V cavities were restored with an ormocer-based resin composite (Admira, Voco). Three restorative techniques were used: conventional bulk insertion (Group I or control group) (n = 53 sections), room-temperature resin composite inserts (Group II) (n = 52) and cooled resin composite inserts (Group III) (n = 50). Microleakage and penetrating microleakage were studied under the microscope. Cooled composite inserts reduce microleakage at the gingival margins with respect to Groups I (p = 0.002) and II (p = 0.014). When small-size cooled composite inserts were used, the sealing ability at the gingival margins of Class V composite restorations was improved with respect to the bulk insertion technique and the room-temperature composite inserts technique. PMID- 12760703 TI - The effect of elevated temperatures on the dentin adhesion of resin composites. AB - Although resin composite restorations may undergo relatively extreme temperature changes in the oral cavity, little is known about the effects of temperature on their adhesion to tooth structure. This study evaluated the effect of temperature on shear bond strength to dentin of three commercial resin dentin adhesives through testing of matured specimens over the 20 degrees to 55 degrees C temperature range. A significant difference (p < 0.05) was observed between 20 degrees C and 55 degrees C for all the materials, and for one of the materials, a significant difference was also observed between 20 degrees C and 37 degrees C. PMID- 12760704 TI - An energy conversion relationship predictive of conversion profiles and depth of cure for resin-based composite. AB - Predicting the polymerization throughout resin-based composite (RBC) has been reduced to a set of variables involving irradiance of the light source, exposure duration and RBC transmission properties, together with an energy-conversion relationship (ECR) derived from Fourier Transform Infrared Spectroscopic analysis (FTIR) of a single shade of photo-polymerized RBC. The ECR describes the localized energy density required to achieve a desired conversion independent of shade. Using this ECR, conversion was predicted and experimentally verified throughout different opacities of RBC based on knowledge of their transmission properties and the incident radiant energy density (irradiance times exposure time). Also, using RBC transmission properties, a critical scrape-back energy of approximately 32 mJcm(-2) was determined from cylindrical samples of photo polymerized RBC in which the poorly polymerized material was removed. This value correlates to approximately 22% conversion. The critical scrape-back energy was then used to predict scrape-back lengths obtained from samples polymerized at various energy densities. These results confirm the logarithmic relationship between depth of cure and energy of exposure and the reciprocal relationship between irradiance and time of exposure. PMID- 12760705 TI - Influence of curing tip distance on resin composite Knoop hardness number, using three different light curing units. AB - This in vitro study evaluated the influence of curing tip distance on the Knoop Hardness Number (KHN) of a resin composite when using three different light curing units: (1) a halogen light (XL 1500 curing unit-3M), (2) a "softstart polymerization" (Elipar Trilight curing in an exponential mode-ESPE) and (3) a PAC (Apolo 95E curing unit-DMD). The resin composite, Filtek Z250 (3M), was cured by these curing units at three light-tip distances from the resin composite: 0 mm, 6 mm and 12 mm. The resin composite specimens were flattened to their middle portion and submitted to 18 KHN measurements perspecimen. The results showed that for the Elipar Trilight unit, the hardness of the resin composite decreased as the light tip distance increased. The XL 1500 unit presented a significant decrease in hardness as the depth of cure of the resin composite increased. Apolo 95E caused a decrease in the resin composite hardness values when the depth of cure and light tip distance increased. PMID- 12760706 TI - Conservative restoration of proximal-cervical lesions. AB - One of the authors has used this technique successfully for the past three years in various clinical situations that involve both difficult access anterior and posterior teeth. This is a tooth-structure conserving clinical procedure that can provide a simplified approach for restoring otherwise difficult clinical lesions. PMID- 12760707 TI - Measurement of colonic transit time in patients with chronic idiopathic constipation. PMID- 12760708 TI - Measuring colonic transit time in chronic idiophatic constipation. AB - INTRODUCTION AND OBJECTIVES: Colonic transit time can define three types of delay: right colon (colonic inertial), left colon and recto sigmoid. The aim of this study is to compare clinic characteristics and manometric results among the different groups with segmental colonic transit disorder. PATIENTS AND METHODS: 61 subjects were included in our study (5 men and 56 women) with and average age of 47.5 years (range: 17-84 years). We carried our a study of segmental and total colonic transit time, with the use of radio-opaque markers and sequential abdominal X-rays. We considered that transit time was prolonged when it was longer than 72 hours, and segmental transit time, when right colon was longer than 22 hours, left colon 37.2 hours and recto-sigmoid 37.2 hours. We included different clinic data: age, sex, place of residence stool frequency, abdominal pain, painful evacuation, incomplete evacuation, evacuation desire, dietary fibre intake, use of laxatives. We practised anorectal manometry in 20 patients. RESULTS: The average total colonic transit time was 38.2 hours in normal transit and 103 hours with disorder. The frequency in the three colonic segments, when there was a long total colonic transit time, was: 40% in the left colon, 33% in the recto-sigmoid and 27% in the right colon. We analysed the clinical characteristics of these three groups, finding more frequency of painful evacuation and defecation desire and lower dietary fibre intake in recto sigmoid. The most important abnormality in anorectal manometry was the hiposensitivity in the anorectal area, that only appeared in subjects with long segmental transit time in the right colon (colonic inertia), statistically significant. CONCLUSIONS: The measurement of colonic transit time with radio-opaque markers is a simple technique that allows for the objective detection of the different groups of patients with chronic idiopathic constipation. The exact typification of the abnormality is important for the individualization of each treatment. PMID- 12760709 TI - Recurrence pattern of esophageal cancer after esophagectomy with two-field lymphadenectomy. AB - AIM: To analyze the pattern of recurrence of esophageal carcinoma after a curative-intention surgical resection. PATIENTS: Ninety-two patients with non metastatic esophageal carcinoma were included. Ninety percent of patients were male, and the mean age of this series was 61 years. The most frequent histologic subtype was squamous cell carcinoma. Fifty percent of tumors were at or above the tracheal bifurcation. All patients were submitted for transthoracic subtotal esophagectomy plus two-field radical lymphadenectomy, leaving no apparent residual disease. No adjuvant therapy was applied to any patient. RESULTS: Follow up was complete for 76 out of 80 patients surviving the operation. Thirty-four tumoral recurrences were detected for a disease-free survival af 39% at 9 years after surgery. All recurrences were detected during the first two years after treatment. Tumoral relapse was related to the presence of T3 or T4 tumors, with positive lymph nodes, squamous cell carcinoma subtype and supracarinal location. Nine percent of patients had a distant relapse, 15% had a locorregional relapse and 12% a combination of both. Distant relapse presented significantly earlier. There was no statistical association between type of recurrence and clinico pathological or surgical features. CONCLUSIONS: After radical surgery for carcinoma of the esophagus, half of the patients relapse in the following two years. Distant metastases happen to appear earlier in the follow-up, but the most frequent recurrence is the locorregional one. PMID- 12760710 TI - Seven-day proton pump inhibitor, amoxicillin and clarithromycin triple therapy. factors that influence Helicobacter pylori eradications success. AB - AIM: To evaluate which factors influence eradication success with standard triple therapy for Helicobacter pylori. PATIENTS AND METHODS: A prospective study was made of 891 patients infected by H. pylori and diagnosed with duodenal ulcer (n=422), gastric ulcer (n=221), or functional dyspepsia (n=248). Initially, an endoscopy with biopsies of antrum and body (haematoxylin-eosin stain), and a 13C urea breath test were performed. All patients were treated for seven days with either omeprazole 20 mg twice daily in 442 patients (OCA) or pantoprazole 40 mg twice daily in 449 patients (PCA), associated to clarithromycin (500 mg twice a day) and amoxicillin (1 g twice a day). Two months after completing therapy urea breath test was repeated to confirm eradication. RESULTS: Mean age +/- SD was 51.6 +/- 15 years, 61% were male. Overall eradication rate was 73.7% (95% CI 69 77%) and 80.8% (77-84%) with OCA and PCA therapy, respectively, showing significant difference between treatment regimens (chi 2 =6.3; p= 0.01). As refers to underlying diseases, H. pylori eradication was achieved in 77.4% (74 80%) of peptic ulcers and 77% (71-82%) of functional dyspepsia (p=n.s.). With our two treatment regimens (OCA/PCA) eradication success was 74/81% in peptic ulcer (p=0.03), and 72/80% in functional dyspepsia (p=0.1). In the multivariate analysis, type of therapy was the only variable that correlated with eradication success (odds ratio 1.5; 95% CI: 1.1-2.1) (chi2 model: 6,4; p=0.01). CONCLUSIONS: Standard triple therapy containing a proton pump inhibitor, clarithromycin and amoxicillin for seven days achieves in our community a moderate eradication success; this result could improve by using pantoprazole instead of omeprazole. This therapy is equally effective in patients with peptic ulcer and functional dyspepsia. PMID- 12760711 TI - Attitude facing a patient suffering from Helicobacter pylori infection resistant to a triple therapy. AB - Even with the current most effective treatment regimens, about 10-20% of patients will fail to eradicate H. pylori infection. Therefore, in designing a treatment strategy we should not focus on the results of primary therapy alone, but also on the final (over-all) eradication rate. The choice of a second-line treatment depends on which treatment was used initially, as retreatment with the same regimen is not recommended. In this respect, the first therapy should not be a regimen that combines clarithromycin and metronidazole in the same regimen, because of the problem of resistance against both antibiotics. Therefore, it seems that performing culture after a first eradication failure is not necessary and assessing H. pylori sensitivity to antibiotics only after failure of the second treatment may be suggested in clinical practice. Different possibilities of empirical treatment are suggested. After failure of proton pump inhibitor (PPI)-amoxicillin-clarithromycin, quadruple therapy has been generally used. More recently, replacing the PPI and the bismuth compound by ranitidine bismuth citrate (RBC) has also achieved good results. After PPI-amoxicillin nitroimidazole failure, retreatment with PPI-amoxicillin-clarithromycin has proved to be effective. Finally, rifabutin-based rescue therapies have shown to constitute an encouraging strategy for eradication failures, as they are effective for H. pylori strains resistant to antibiotics. PMID- 12760712 TI - A continuous spectrum of neutrophilic dermatoses in Crohn's disease. AB - The inflammatory bowel disease is accompanied by cutaneous manifestations in approximately 10% of the cases. Neutrophilic dermatoses are located on the dermis and/or epidermis and are characterised on histological examination by the presence of an infiltrate that consists largely of neutrophils. The prototype of neutrophilic dematoses is Sweetacute;s syndrome; which is rarely associated with Crohns disease. CASE REPORT: A 63 year old woman was admitted to hospital with pyrexia, abdominal pain, episcleritis and skin lesions. She presented erythematous lesions on trunk, legs and arms, with tendency towards formation of plaques, nodules and vesicular pustular lesions. Both the colonoscopy and colonic biopsies confirmed the diagnosis of colonic Crohns disease. Cutaneous biopsies re vealed an infiltrate consisting mainly of neutrophils. These biopsies, together with clinical details led to the diagnosis of Sweetacute;s syndrome. A methylprednisolone treatment rapidly improved the skin lesions and clinical symptoms. The different clinical forms of neutrophilic dermatosis are an extra intestinal manifestation of Crohns disease, and are some-times found concurrently in the same patient, which would indicate a common pathogenesis with different clinical presentations (spectrum of neutrophilic dermatoses). PMID- 12760713 TI - [Acute acalculous cholecystitis by Escherichia colli in the intensive care unit]. PMID- 12760714 TI - [Similar poor response in genotype 4 chronic Hepatitis C treatment with interferon and ribavirin compared with genotype 1 treatment]. PMID- 12760715 TI - [Is intestinal anisakiasis a frequent infestation?]. PMID- 12760716 TI - Gastric carcinogenesis and Helicobacter pylori infection. PMID- 12760717 TI - Helicobacter pylori infection affects the expression of PCNA, p53, c-erbB-2 and Bcl-2 in the human gastric mucosa. AB - OBJECTIVE: Helicobacter pylori infection has been related to gastric carcinogenesis. This association is based on epidemiological data, pathological changes observed in the gastric mucosa, and chemical products from bacteria that may induce damage of DNA. In the present study we examined gastric endoscopic biopsies from patients with chronic gastritis, with and without H. pylori infection, and surgical biopsies from gastric cancer patients to evaluate whether this bacteria may induce changes in the expression of molecular markers associated with carcinogenesis. PATIENTS AND METHODS: the study involved 57 biopsies from the antral region of the stomach of patients with chronic gastritis and gastric cancer that were analyzed by immunohistochemistry. Molecular markers examined were: PCNA (Proliferating Cell Nuclear Antigen), p53, c-erbB-2, Bcl-2, and p21 H-ras. RESULTS: PCNA content of epithelial cells was significantly higher in H. pylori infected biopsies. Treatment aimed to eradicate H. pylori decreased the level of PCNA-positive cells in the group of patients that became H. pylori negative as well as in H. pylori-positive patients. Nuclear p53 expression (used here as a surrogate marker for p53 mutation/inactivation) and c-erbB-2 expression were observed only in the group of patients that remained with the bacteria after treatment. A higher bcl-2 expression in lymphoid cells was observed in H. pylori positive biopsies, and treatment did not change the expression of this protein. No significant expression of p21 H-ras was observed in the studied biopsies. CONCLUSION: this study suggests that H. pylori is involved in the induction of molecular changes that might predispose human gastric mucosa cells to pre neoplastic and neoplastic events. PMID- 12760718 TI - Effectiveness of rubber band ligation in haemorrhoids and factors related to relapse. AB - PURPOSE: to assess the effectiveness of ambulatory rubber band ligation (RBL) in the treatment of symptomatic internal haemorrhoids and to identify factors related to relapse. PATIENTS AND METHODS: prospective study of 232 patients treated with rubber band ligation for symptomatic haemorrhoids (grade I-III or grade IV with severe contraindication for surgery) from November 1996 to November 2000 at the outpatient clinic. Ligation was performed with a Stille AB (Comedic) ligator and suction pump, placing 1-3 bands per session and with up to three sessions per patient. Effectiveness of treatment was defined as the absence of symptoms and was confirmed by anoscopy by checking the residual scar after the cushions' detachment. Categorical variables were compared using the shi-squared test, whereas Student's t-test was used for continuous variables. Logistic regression was employed to identify clinical factors related to relapse. RESULTS: a total of 331 bands were placed during 235 sessions in the 163 patients who completed follow-up (70%). Mean age was 45.6 years, with males accounting for 64.4%. Most patients (86.5%) had grade II or grade III haemorrhoids. Overall morbidity was 6%. The most frequent complications were rectal tenesmus (11%), slight or mild anal pain (7.4%), dysuria (4.3%) and transient anal bleeding (3.7%). The treatment was effective in 86% of patients after a mean follow-up of 32 months. Efficacy was high for grades I and II (100% and 97.4% ) but decreased for grade III (69.8%; p<0.001) and grade IV (0%; p<0.001). Most relapses occurred within the first 24 months (87%) and were not significantly related to age, gender, duration of symptoms, itching, bleeding, pain, tenesmus or bowel habit, but were significantly related to the presence of prolapse and its grade (p<0.001), and to the involvement of left posterior, right lateral and anterior pedicles (p<0.05). CONCLUSIONS: ambulatory RBL is a safe and effective treatment for grade I, II and III symptomatic haemorrhoids, and is associated with low morbidity. Recurrence is uncommon and occurs mainly within the first 24 months, being related to the presence and grade of prolapse as well as to its location, but bears little relation to the rest of factors analysed. PMID- 12760719 TI - Validation of the 13c-urea breath test for the initial diagnosis of helicobacter pylori infection and to confirm eradication after treatment. AB - OBJECTIVES: the breath test with 13C-urea (UBT) is a method widely used in Spain, but its diagnostic accuracy has not been evaluated in a clinical trial until now. Our objective was to validate the UBT (TAU-KIT) both as an initial diagnostic method for the detection of H. pylori infection and as a method to confirm eradication. METHODS: a multi-centre study in 7 Spanish hospitals was performed. A group of dyspeptic patients who had not previously received eradication treatment was included, and a second group of patients with gastric ulcer or upper gastrointestinal bleeding due to peptic ulcer was also included (eradication of H. pylori was confirmed 6 to 8 weeks after treatment completion with omeprazole, clarithromycin and amoxycillin). In both groups an endoscopy was performed with biopsies for histology and rapid urease test. Patients were considered infected if both tests yielded positive results, and not infected when both tests were negative. The UBT 13C-urea (TAU-KIT, Isomed S.L., Madrid, Spain) was performed with citric acid and 100 mg of 13C-urea. The pathologist and persons responsible for endoscopy, urease test and UBT were all unaware of the results from the other diagnostic methods. RESULTS: in the pre-treatment group (36 patients) the prevalence of H. pylori was 72%, the area under the ROC curve for the diagnosis of infection with the UBT was 0.99, and the best cut-off point was 5 units, with the following results: sensitivity= 96% (95% CI = 81-99%), specificity= 100% (69-100%), positive predictive value (PPV) = 100% (87-100%), negative predictive value (NPV) = 92% (59-100%), likelihood ratio (LR) + = infinity, and LR- = 0.04. In the post-treatment group (85 patients) the prevalence of H. pylori was 16%, the area under the ROC curve was 0.99, and the best cut point was 4.6, with the following results: sensitivity= 100% (77-100%), specificity = 97% (90-99%), PPV = 88% (62-98%), NPV = 100% (95-100%), LR+ = 35, and LR- = 0. CONCLUSION: UBT provides excellent accuracy both for the initial diagnosis of H. pylori infection and to confirm eradication after treatment. PMID- 12760720 TI - Hepatic encephalopathy: nomenclature, pathogenesis and treatment. AB - Hepatic encephalopathy (HE) is a neuropsychiatric syndrome in patients with liver failure and/or a portal-systemic bypass. Since 2002 a new nomenclature of HE exists, that classifies HE in encephalopathy type A (associated with acute liver failure), type B (associated with portal-systemic bypass), and type C (associated with liver cirrhosis). HE type A is characterized by a rapid development to coma, cerebral edema, and a poor short-term prognosis. Therefore, these patients should be referred to a liver transplantation center. Standard treatment of HE consists of non absorbable disaccharides, non absorbable antibiotics, and a diet with an appropriate amount of proteins. In addition, the possibility of performing a liver transplantation should be evaluated. In patients with intractable HE other alternative treatments adjunct to standard treatment, like zinc, sodium benzoate, ornithine aspartate, branched chain amino acids, flumazenil, and bromocriptine should be considered. PMID- 12760721 TI - Whipple's disease: report of cases diagnosed in our hospital. AB - We present here a retrospective study of nine patients diagnosed with Whipple's disease (WD) in our hospital. This report em-phasises clinical presentation, diagnostic methods, treatment and response to treatment. In our study, the disease was more frequent in males, and the most frequent presenting symptoms were arthralgia, diarrhoea and weight loss. Since the intestine is almost always affected, oral endoscopy is a useful technique for the diagnosis of WD because it shows the typical miliary pattern and aids in obtaining biopsies to show the presence of PAS-positive macrophages (a suggestive though not diagnostic finding), to show bacilli using electron microscopy, or to detect genetic material using PCR. Our patients responded well to treatment. The most frequently used antibiotic was oral trimethoprim-sulfamethoxazole for at least one year. Treatment with penicillin G and IM streptomycin for 14 days was reserved for severe cases or cases that responded poorly to treatment. PMID- 12760722 TI - Polyarteritis nodosa and hepatitis B virus infection. PMID- 12760723 TI - [Splenic epidermoid cysts]. PMID- 12760724 TI - [Hydro-saline depletion syndrome secondary to villous adenoma of the rectum: mechanism of secretion. A case report]. PMID- 12760726 TI - Therapeutical alternatives in achalasia. PMID- 12760727 TI - Short- and medium-term clinical efficacy of three endoscopic therapies for achalasia: a single-blinded prospective study. AB - OBJECTIVE: to compare the efficacy of three endoscopic therapies for achalasia and to identify predictors of response. DESIGN: prospective, single-blinded study at short and medium term. MATERIAL AND METHODS: 22 patients (9M/13 F; mean age: 47.45 +/- 21.01 years) with confirmed clinical and manometric achalasia were randomised in three groups: intrasphincteric injections of botulinum toxin (group 1: 10 patients ), injections of 1% polidocanol (group 2:6 patients), and a combined therapy with both of them (group 3: 6 patients). Clinical response was evaluated by a score (0-5) of tested symptoms (dysphagia, regurgitation and chest pain) at 1 and 24 weeks post-treatment. RESULTS: at 24 weeks post-treatment group 2 had the best complete response (CR) rate (33.33%), whereas CR in both the botulinum toxin and combined therapy groups was 10 and 0%, respectively. Groups 1 and 2 got an overall improvement in clinical score at 1 (p= 0.02) and 24 weeks (p= 0.04). Five patients (50%) in group 1, two patients (33.33%) in group 2, and three patients (50%) in group 3 needed other therapies (dilation or surgery) because of treatment failure. Separately, neither age nor sex, time from diagnosis or type of therapy could distinguish responders from non-responders in these three groups. However, absence of response within the first week, and an initial clinical score above 7 were predictive factors of poor response at six months. CONCLUSIONS: short- and medium-term clinical response to these endoscopic therapies was limited. The absence of response at seven days and a severe initial clinical score were predictive factors of poor medium-term response. PMID- 12760728 TI - Acute typhlitis in inmunocompromised patient: an eight year experience. AB - INTRODUCTION: Acute typhlitis is usually associated with severe immunosuppressive conditions. Initially described as closely associated with infantile myeloid leukaemia, its incidence increased along the last decade. DESIGN: retrospective review. PATIENTS: 12 immunodepressed patients affected of acute typhilis in our hospital between 1994 and 2001. Suspected diagnosis was established by clinical symptoms and abdominal CT findings, and was confirmed with pathological finding in the surgical specimen. Clinical and radiological diagnosis, treatment, complications and survival of patients are discussed. RESULTS: 3 patients with a previous diagnosis af acute myeloid leukemia, 2 patients with non-Hodgkin lymphoma, 2 patients with aplastic anaemia, one patient with AIDS, and 4 patients with kidney transplantation were included in our study. Prednisone, cyclosporine, Ara-C and vincristine were the most frequently involved drugs. Most frequent clinical findings included abdominal pain, fever, nausea-vomiting and abdominal distension. CT diagnosis revealed caecum and colic involvement with rarefaction of pericaecal fat. Medical treatment was successful in only 33% of all patients, the other patients requiring a surgical procedure including right hemicolectomy with or without intestinal anastomosis. Mortality reached 58.3 per cent, representing multiorganic sepsis the main cause of death. CONCLUSIONS: although early diagnosis of acute typhlitis bears a better prognosis, mortality rates are up 50 % in spite of an established treatment. PMID- 12760730 TI - Transendoscopic ultrasound mini-probes, are they necessary? AB - The goal of this paper was to define the current state of mini-probes or miniaturized endoscopic ultrasound probes, their technical specifications, current clinical applications, safety and minimal risks, complications and limitations. PMID- 12760729 TI - Acute pancreatitis in childhood. AB - AIM: to evaluate etiological agents involved in acute pancreatitis in children, as well as clinical, laboratory and radiological findings and the illness clinical course. PATIENTS AND METHODS: we reviewed the cases of acute pancreatitis diagnosed over the last 15 years. The criteria used for cases to be included were acute abdominal pain, elevated serum amylase levels and/or ultrasound abnormalities in the pancreatic area in conscious patients, and the last two criteria in unconscious patients. RESULTS: thirty-one children were included (average age 7.9 years, range 2-15; 55% males). Infection and gallstones were the most common causes (19 and 16 %, respectively). In all, 9.7% of cases were drug-related (valproic acid, L-asparaginase, azathioprine combined with high doses of methylprednisolone); 6.5% were traumatic in origin and another 6.5% was due to systemic diseases. In 35.5 % no cause was found (idiopathic). The most frequent symptoms were abdominal pain (90%) and emesis (38%). Amylase serum levels were elevated in all patients. Abdominal ultrasound scans were abnormal in 64%, with an increase in the pancreatic area in 48% and hypoechogenicity in 51%. Seven cases required surgical treatment (22%). Seven children had acute pancreatitis, and three of them died as a result of shock unrelated to pancreatitis. Relapse of disease occurred in 19% of patients. CONCLUSIONS: acute pancreatitis should always be considered in children with abdominal pain. There are a wide variety of etiological factors and in a high percentage of patients no underlying cause is found. Prognosis is variable owing to the heterogeneity of the clinical course of this illness in children. PMID- 12760731 TI - D-penicillamine and plasmapheresis in acute liver failure secondary to Wilson's disease. AB - We report a case of a 19-year-old woman with acute liver failure, Coombs negative hemolytic anemia, and renal failure as initial manifestations of Wilson disease with recovery following medical treatment. The clinical picture and low serum transaminase and alkaline phosphatase levels gave us a clue to suspect Wilson disease and to initiate plasmapheresis and D-penicillamine soon after admission. The serum and urinary copper levels were elevated with low serum ceruloplasmin. We proceeded to ambulatory follow-up with medical treatment with D-penicillamine. A few months later, during the course of a laparoscopic cholecystectomy because of symptomatic gallstone disease, a liver biopsy sample was obtained that showed histological liver fibrosis and strongly elevated levels of liver tissue copper. PMID- 12760732 TI - [Splenic infarction as a complication of atrial fibrillation]. PMID- 12760733 TI - [Acute pancreatitis and primary hyperparathyroidism]. PMID- 12760734 TI - [Necrotizing fasciitis and streptococcal toxic shock syndrome after hemorrhoidectomy]. PMID- 12760742 TI - Expression and coordinated regulation of matrix metalloproteinases in chronic hepatitis C and hepatitis C virus-induced liver cirrhosis. AB - Matrix metalloproteinases (MMPs) are central to tissue remodelling; however, little is known about the temporal pattern and differential regulation of hepatic MMP expression in the course of chronic human liver disease. Using quantitative reverse transcription-PCR ELISA assays, we studied hepatic mRNA expression of MMP 1, -2, -3, -7, -9, -10, -11, -13 and -14 in patients with chronic hepatitis C and hepatitis C virus-induced end-stage liver cirrhosis and controls. Results were compared with histology, hepatic expression of tissue inhibitor of metalloproteinases (TIMP)-1, -2 and -3, procollagen types I and IV, laminin, and with circulating protein levels of hyaluronate, TIMP-1 and -2 and MMP proenzymes, as measured by ELISA. The impact of the MMP-3(-1171) promoter polymorphism on hepatic MMP-3 expression was analysed. Hepatic mRNA expression data identified differentially regulated groups of MMPs during the course of chronic hepatitis C, showing either steadily increasing mRNA expression with disease progression (MMP 1, -2, -7 and -14) or transiently elevated expression (MMP-9, -11 and -13). The first group closely correlated to the parameters of fibrogenesis. Hepatic MMP-3 expression was unrelated to disease stage, but was determined by the MMP-3(-1171) promoter polymorphism. In conclusion, MMP expression during the course of chronic hepatitis C appears to be a closely regulated process, with different clusters of coordinately regulated MMP genes being identified. PMID- 12760743 TI - Cloning of an emopamil-binding protein (EBP)-like protein that lacks sterol delta8-delta7 isomerase activity. AB - EBP (emopamil-binding protein) is a high-affinity binding protein for [3H]emopamil and belongs to the family of so-called sigma receptors. Mutations that disrupt EBP's 3beta-hydroxysteroid sterol delta8-delta7 isomerase activity (EC 5.3.3.5) impair cholesterol biosynthesis and cause X-chromosomal dominant chondrodysplasia punctata. We identified a human cDNA for a novel EBPL (EBP-like protein) with a calculated mass of 23.2 kDa. Amino acid sequence alignments and phylogenetic analysis revealed that EBPL is distantly related to EBP (31% identity and 52% similarity) and found in animals but not in plants. EBPL is encoded by four exons on human chromosome 13q14.2 covering 30.7 kb, and a partially processed EBPL pseudogene was found on 16q21. The EBPL mRNA was expressed ubiquitously and most abundant in liver, lung and kidney. Upon heterologous expression in yeast EBPL had no detectable 3beta-hydroxysteroid sterol delta8-delta7 isomerase and sigma-ligand-binding activity. Nine out of ten amino acid residues essential for catalytic activity of EBP were conserved in EBPL. Replacement of the only differing residue (EBP-Y111W) reduced catalytic activity of EBP. Transfer of the divergent residue from EBP to EBPL (EBPL-W91Y) and chimaerization of EBP and EBPL at various positions failed to restore catalytic activity of EBPL. Chemical cross-linking induced homodimerization of EBPL and EBP. Whereas mevinolin increased the mRNA for EBP and DHCR7 (delta7 sterol reductase) in HepG2 cells, it had no effect on mRNAs for EBPL and sigma1 receptor, indicating that EBP and EBPL expression are not co-ordinated. We propose that EBPL has a yet-to-be-discovered function other than cholesterol biosynthesis. PMID- 12760744 TI - A novel method for the immobilization of tyrosinase to enhance stability. AB - A new method of tyrosinase immobilization by Fuller's-earth adsorption followed by entrapment in gelatin has been developed with 98% activity immobilization yield. An appreciable increase in operational and thermal stability was observed for FEAGE (Fuller's earth-adsorbed gelatin-entrapped) tyrosinase compared with GE (gelatin-entrapped) native enzyme. FEAGE tyrosinase could be used repeatedly after intermittent storage and retained 70% of its initial activity after eight cycles. The half-life of the GE enzyme at 40 degrees C was less than 2 h, whereas the FEAGE enzyme retained about 75% of its initial activity after 2 h. Taken together our data demonstrate clearly that the technique of immobilizing tyrosinase via adsorption followed by entrapment appears promising and is hence recommended for tyrosinase immobilization for commercial production of L-DOPA (3,4-dihydroxyphenylalanine). PMID- 12760745 TI - Identification of conserved regulatory elements by comparative genome analysis. AB - BACKGROUND: For genes that have been successfully delineated within the human genome sequence, most regulatory sequences remain to be elucidated. The annotation and interpretation process requires additional data resources and significant improvements in computational methods for the detection of regulatory regions. One approach of growing popularity is based on the preferential conservation of functional sequences over the course of evolution by selective pressure, termed 'phylogenetic footprinting'. Mutations are more likely to be disruptive if they appear in functional sites, resulting in a measurable difference in evolution rates between functional and non-functional genomic segments. RESULTS: We have devised a flexible suite of methods for the identification and visualization of conserved transcription-factor-binding sites. The system reports those putative transcription-factor-binding sites that are both situated in conserved regions and located as pairs of sites in equivalent positions in alignments between two orthologous sequences. An underlying collection of metazoan transcription-factor-binding profiles was assembled to facilitate the study. This approach results in a significant improvement in the detection of transcription-factor-binding sites because of an increased signal-to noise ratio, as demonstrated with two sets of promoter sequences. The method is implemented as a graphical web application, ConSite, which is at the disposal of the scientific community at http://www.phylofoot.org/. CONCLUSIONS: Phylogenetic footprinting dramatically improves the predictive selectivity of bioinformatic approaches to the analysis of promoter sequences. ConSite delivers unparalleled performance using a novel database of high-quality binding models for metazoan transcription factors. With a dynamic interface, this bioinformatics tool provides broad access to promoter analysis with phylogenetic footprinting. PMID- 12760746 TI - [Prevention, diagnosis and treatment of biliary complications after liver transplantation]. AB - OBJECTIVE: To study the prevention, diagnosis and treatment of biliary complications after liver transplantation. METHODS: One hundred and twenty-three recipients who had received liver transplantation between April 1993 and November 2001 were retrospectively reviewed. RESULTS: Biliary complication was diagnosed by cholangiography in 11 patients. Nine patients were cured, 1 patient was improved and 1 patient died. The incidence for biliary complications was 8.9% (11/123), and the mortality was 0.8% (1/123). The incidence of biliary complications due to T tube was 4.2% (5/119). The incidence of biliary complications due to hepatic artery was 1.6% (2/123). In recipients with WIT > 3 min and CIT > 8 h, the incidence of biliary complications elevated significantly. CONCLUSIONS: The most important reason for biliary complications was preservative and ischemic injury. While repairing donor liver, damage to the blood supply system of donor liver bile ducts should be avoided. Meticulous T tube placement may reduce the incidence of biliary complications. Early cholangiography is helpful to diagnose biliary complications. PMID- 12760747 TI - [Clinical analysis of different periods of liver transplantation at an organ transplantation centre]. AB - OBJECTIVE: To summarize our clinical experience in liver transplantation while considering the background in this field in China. METHODS: Ninety-five patients who had received liver transplantation from April 1993 to March 2002 were analyzed retrospectively. Three periods were defined objectively as period I (1993 - 1997), II (1999) and III (2000 - 2002). Operative techniques, recipients, original diseases, complications and survival rates were compared among the three periods. RESULTS: Malignant liver lesions were the main cause for liver transplantation in period I and II. The ratio of number of malignant disease to total recipients decreased gradually from period I to III (100%, 53% and 35%, respectively). The 1-year survival rate in patients with benign liver disease was 85% and the total operative mortality was 5% in period III. The incidence of hepatitis B virus reactivation or reinfection was 24% twelve months after liver transplantation. Vascular complication decreased but biliary complications did not and remained a major long-standing problem. No veno-venous bypass technique was used in period III, and its advantages were obvious when comparing with those with veno-venous bypass in period I and II. CONCLUSIONS: Strict selection of recipients, fine operative technique, familiarity with various complications and correct therapeutic methods, prophylaxis of recurrence of hepatitis B and hepatocellular carcinoma are necessary to improve long-term results of liver transplantation in China. PMID- 12760748 TI - [Venovenous bypass in 54 liver transplantations]. AB - OBJECTIVES: To summarize hemodynamic and metabolic changes during bypass, and to evaluate the bypass in liver transplantation. METHODS: Fifty-four patients underwent orthotopic liver transplantation with venovenous bypass from May 2000 to May 2002. Their clinical features were analysed. RESULTS: SHR, MAP, CVP, CO, PaO(2), PaCO(2), serum K(+), Na(+), Ca(2+), BUN values were not significantly changed during bypass. Compared to the pre-bypass stage, pH was decreased in the post-bypass stage (P < 0.05), serum lactic acid value was increased in the bypass and post-bypass stage (P < 0.05), active clotting time was increased in the bypass stage (P < 0.05), serum creatinine value was increased on first postoperative day (P < 0.05). CONCLUSIONS: Venovenous bypass could improve hemodynamic and metabolic stability in the anhepatic phase, but it also could increase operation duration, liver ischemic time and cost. PMID- 12760749 TI - [Some principal surgical techniques for living donor liver transplantation]. AB - OBJECTIVE: To investigate some principal surgical techniques of living donor liver transplantation (LDLT). METHODS: Eleven patients of LDLT have been performed at our department from January 2001 to March 2002. The left lobe (segments II, III, IV, including the middle hepatic veins) was transplanted in 8 patients, the left lateral lobe (segments II, III) in one and the right lobe (segments V, VI, VII, VIII, not including the middle hepatic veins) in 2. The plane of liver resection was determined on the basis of donor liver volumetry using CT scan and the anatomic analysis of vascular structure of the hepatic vein, portal vein and hepatic artery using intraoperative ultrasound. The hepatic parenchyma was transected using ultrasound aspirator without blood vessel clamping or graft manipulation. The isolated graft was perfused in situ through the portal vein branch. The liver graft was transplanted into the recipients who underwent total hepatectomy with preservation of the inferior vena cava. The hepatic vein reconstruction was performed in end to end fashion or end to side to the vena cava after venoplasty. Arterial anastomoses were performed using microsurgical technique. Biliary reconstruction was made by using duct-to-duct anastomosis and placement of a T tube. RESULTS: All the 11 donors are uneventfully discharged after operation. In the 11 recipients, an 8-year-old girl needed retransplantation because of hepatic artery thrombosis, one case died of serious chronic rejection on the postoperative day 72. Ten recipients recovered and were discharged from hospital, whose liver function and cuprum oxidase had returned to normal. CONCLUSIONS: The procedure of LDLT is relatively safe for the donor. Reconstruction of vessels is a key step in the procedure. Comprehending anatomical variation of vessels pre- and intra-operatively and correct surgical management might reduce the incidence of complications. PMID- 12760750 TI - [Diagnosis and treatment of lung aspergillosis after liver transplantation]. AB - OBJECTIVE: To assess the diagnosis and treatment of invasive lung aspergillosis after liver transplantation. METHODS: Routine sputum culture was performed. Itraconazole and fluconazole were used to prevent fungal infection prophylactically. Amphyotericin B was only used on aspergillosis. In 54 patients receiving, liver transplantation, 3 patients with lung aspergillosis were reviewed. RESULTS: Of the 3 patients 2 died and 1 recovered. CONCLUSIONS: Over immunosuppression is a main risk factor for aspergillosis. Amphotericin B is still the best choice for the treatment of aspergillosis and its gradual, interrupted, low concentration administration, cooperated with itraconazole can ease the side effects. PMID- 12760751 TI - [The role of IL-15 in the process of acute rejection in liver and heart transplantation in rats]. AB - OBJECTIVE: To study the real effect of IL-15, a kind of T lymphocyte activators which were derived from lymphocytes, on the acute rejection process in heart and liver transplantation in rats. METHODS: Male (body weight 200 - 250 g) 1A (RT1(a)) and LEW (RT1(l)) rats were selected as donors and recipients, respectively. Heterotopic heart transplantation (in abdomen) and orthotopic liver transplantation were performed as the acute rejection model group (experimental group); LEW (RT1(l))-->LEW (RT1(l)) as donors and recipients to establish isografts transplantation as the control group. Animals were sacrificed on day 1, 3, 5, 7 and graft specimens were collected. Microarray, immunohistochemistry and Western-blotting methods were used to detect the expressions of IL-15, IL-2 and IFN-gamma, etc. 48 rats were divided evenly into two groups and each time-point consisted of 6 rats. RESULTS: Acute rejections which were clarified by pathological findings and animal manifestations were found 3 days after operation in the experimental group. The early expression of IL-15 was found on endothelial cells in allografts 1 day after operation in contrast to IL-2, which expressed lately and only be found on inflammatory cells including lymphocytes and Kupffer cells 3 days after graft implantation. The result of INF-gamma was the same as that of IL-2. CONCLUSIONS: IL-15 appeared earlier in heart and liver allografts than IL-2 and IFN-gamma in rat acute rejection model, and the expression site differed from the later two. IL-15 participated in acute rejection reaction earlier in this process and the pathway may be different from IL-2 and IFN-gamma. Early blocking this pathway combined with other blockade would have a promising result in control of the progression of acute rejection. PMID- 12760752 TI - [Application of ultrasound-guided percutaneous radiofrequency ablation in treatment of liver cancer]. AB - OBJECTIVE: To investigate the therapeutic efficacy and its influencing factors of ultrasound-guided percutaneous radiofrequency ablation (PRFA) in the treatment of liver carcinoma. METHODS: With a temperature-controlled multi-electrode needle, ultrasound-guided PRFA was employed to treat forty-seven patients with 67 tumor nodules, with a diameter of 2.6 +/- 1.1 cm (1.0 - 5.5 cm). RESULTS: A complete ablation (CA) rate of 80.6% was achieved in the present series, with a CA rate of 91.7% in the tumors < or = 3 cm in diameter, 75.0% in tumors from 3.1 to 4.0 cm, and 14.3% in tumors > 4 cm. The CA rate was significantly greater in tumors with a temperature rising up to 70 degrees C within the initial 2 minutes at ablation as compared with that longer than 2 minutes (P < 0.05). A markedly higher CA rate was obtained in tumors with an ablation-maintaining temperature of over 80 degrees C than that between 70 degrees C and 80 degrees C (P < 0.01). All patients were followed up with a mean time of 11.3 months. The local recurrence rate was 9.3% (5/54), and 1-year survival rate was 82.1%. Eighteen patients (38.3%) had a distant recurrence. CONCLUSIONS: The tumor size, temperature-rising time and ablation-maintaining temperature represented the important factors affecting the therapeutic efficacy of PRFA. PMID- 12760753 TI - [Clinical values of palliative gastrectomy for late-staged gastric cancer]. AB - OBJECTIVE: To investigate the clinical importance of palliative gastrectomy for late-staged gastric cancer. METHODS: From June 1994 to October 2001, 95 patients with late-staged gastric cancer underwent palliative operation. Clinicopathological and prognostic parameters between 64 patients with palliative gastrectomy (PG group) and 31 patients with unresectable operation (UO group) was compared retrospectively. RESULTS: The age and gender ratios were not different between the two groups. The incidence of large volume (diameter > or = 8 cm), serosal invasion (T(4)) and late TNM stage (IV stage) were significantly higher in the UO group than that in the PG group. There was no difference in peritoneal dissemination, distant lymph node and hepatic metastasis, and tumor location between the two groups. The one- and two-year survival of the patients in the PG group was 48.1% and 23.1%, and significantly better than 13.5% and 0 in the UO group. CONCLUSIONS: Palliative gastrectomy, compared with unresectable operation, can improve the prognosis of the patients with late-staged gastric cancer even with peritoneal dissemination, distant lymph node and hepatic metastasis, and surrounding organ invasion. PMID- 12760754 TI - [Diagnosis and surgical treatment of primary duodenal carcinoma]. AB - OBJECTIVE: To investigate the early diagnosis of primary duodenal carcinoma and its outcome after surgical procedure. METHODS: Twenty-two patients with primary duodenal carcinoma treated operatively between 1983 and 1997 were analyzed retrospectively. Eleven patients complained of epigastric pain and discomfort, 5 jaundice, 4 nausea and vomiting and 2 epigastric fullness. The correct diagnosis rate for endoscopy was 90.0% (9/10), for duodenography 86.7% (13/15), for ultrasound examination 33.3% (4/12) and for computerized tomography (CT) scanning 58.3% (7/12), respectively. All of the 22 patients received surgery including pancreaticoduodenectomy (12 patients), segmental resection (4) and bypass operation (gastrojejunostomy and cholecystojejunostomy or cholangiojejunostomy) (6). RESULTS: Primary duodenal carcinoma was characterized nonspecifically, and the correct diagnosis was based on endoscopy and duodenography. The follow-up rate of this group was 86.4% (19/22). The 1-, 3-, 5-year survival rates of patients receiving radical resection (n = 16, 1 patient lost follow-up) were 86.7% (13/15), 46.7% (7/15), and 26.7% (4/15), respectively. The 5-year survival rate of patients receiving pancreaticoduodenectomy was 27.3% (3/11), and the 5 year survival rate of patients having segmental resection was 1/4. No patient (n = 6, 2 lost follow-up) with primary duodenal carcinoma treated by bypass procedure survived more than one year. There was a significant difference between patients receiving radical procedure and bypass operation (chi(2) = 6.84, P < 0.01). CONCLUSION: Radical resection might improve the survival of patients with primary duodenal carcinoma. PMID- 12760755 TI - [Surgical treatment of degenerative scoliosis]. AB - OBJECTIVES: To investigate the techniques of posterior lumbar interbody fusion and pedicle screws in the treatment of degenerative scoliosis and evaluate their clinical significance. METHODS: Twenty five patients with degenerative scoliosis were treated operatively during 1997 - 2002. Among the 25 patients, 13 patients were treated with Brantigan I/F carbon cages in 28 intervertebral spaces, 7 patients were treated with Prospace cages in 15 spaces, 1 patient was treated with CONTACT cages in 2 spaces. The correcting effects, complications and follow up results were analyzed retrospectively. RESULTS: The average correction rate of scoliosis was 59% and the rate of pain relief was 88%. There was a case of pseudoarthritis and a case of embolism. During follow-up, correction rate and height of disc spaces were not lost nor shift of interbody cages. CONCLUSION: Posterior pedicle screws combined with interbody fusion is a safe and effective treatment for degenerative scoliosis. PMID- 12760756 TI - [Early diagnosis and treatment of pulmonary embolism after total joint replacement: report of five cases]. AB - OBJECTIVE: To study the early diagnosis and treatment of pulmonary embolism (PE) after total joint replacement (TJR). METHODS: >From April 1987 to December 2001, we performed 1,336 total knee replacements (TKR(s)) in 926 patients and 1,745 total hip replacements (THR(s)) in 1,566 patients. In this group there were 5 PE patients after operation. Two patients died (all after TKR), and 3 patients salvaged successfully (2 after TKR, 1 after THR). RESULTS: The total incidence of PE after TJR was 0.2% (5/2,492), the incidence of PE after TKR was 0.4% (4/926), and the incidence of PE after THR was 0.06% (1/1,556). In the patients who suffered PE, 2 died. The incidence of PE before 2000 was 0%. CONCLUSIONS: Death after total joint replacement is due to pulmonary embolism (PE), especially massive PE. The incidence of PE after TKR is higher than that after THR. Pulmonary angiography is the gold standard for the diagnosis of PE. With more understanding on PE and more popularity of joint replacement, the diagnosis of PE after TJR is increased. More attention should be paid to PE in the patients with high risk after TJR. PMID- 12760757 TI - [Congenital coronary artery fistulae in adults: diagnosis and surgical treatment]. AB - OBJECTIVE: To summarize the clinical characteristics, diagnosis and surgical in treatment results of congenital coronary artery fistulas (CAF) in adults. METHODS: Fourteen patients (8 men, 6 women), aged from 18 to 60 years with a mean of 32 +/- 13 years, underwent surgical correction of CAF between March 1985 and April 2002. Eleven of the 14 patients (78.57%) were symptomatic. The diagnosis of CAF was made by echocardiography or angiocardiography preoperatively. The fistulae originated from the right, left and double coronary arteries in 10 (71%), 3 (21%) and 1 (7%) patient(s), respectively. The fistulae drained into the right ventricle (8 patients), left ventricle (4), right atrium (1) and pulmonary artery (1), respectively. The diameter of fistulae ranged from 0.30 to 1.80 cm with a mean of (1.16 +/- 0.49) cm. There were 6 CAF patients associated with coronary artery aneurysms and 4 CAF patients with other coexisting cardiac defects. The distal fistulae were closed in 10 patients with cardiopulmonary bypass (CPB) and 4 patients without CPB. The coexisting defects were corrected simultaneously. RESULTS: There was no early and late death. One patient had low cardiac output syndrome and cured during early postoperative period. Twelve patients (85.71%) were followed up for a mean period of 3.35 +/- 4.28 years without myocardial ischemia or infarction and recurrent fistulae. Heart function was improved to NYHA functional class I in 11 patients and class II in 1 patient. CONCLUSIONS: All adult patients with CAF who have demonstrable hemodynamic and cardiovascular morphological changes should be surgically treated as early as possible. The appropriate surgical management and reliable myocardial protection are key points of good surgical results. PMID- 12760758 TI - [Long-term result of dynamic cardiomyoplasty]. AB - OBJECTIVE: To evaluate the long-term outcome of dynamic cardiomyoplasty in patients with dilated cardiomyopathy. METHODS: Three male patients with dilated cardiomyopathy at age of 36, 45 and 62 years were underwent dynamic cardiomyoplasty from 1994 to 1995. Postoperative follow-up were performed. The data of hemodynamic parameters, functional grade and dimension of heart were analyzed. RESULTS: There was no operative death in these patients. All patients recovered eventlessly. From early postoperative stage, the hemodynamic parameters and clinical symptoms were improved and the patients' exercise tolerance augmented. The quality of life of patients was enhanced significantly. All patients showed NYHA functional grade I. One patient died of severe arrhythmia at the 19th postoperative month. In 2 long-term survivors, the hemodynamic parameters and heart function were declined and the heart tended to be dilated after 3 postoperative years. One patient died of congestive heart failure at 4.5 postoperative years. One patient still survived 7 years after operation and remained heart functional grade II. CONCLUSIONS: Dynamic cardiomyoplasty can significantly improve the functional class and quality of life in patients with dilated cardiomyopathy within a considerable long postoperative period. PMID- 12760759 TI - [Clinical features of postoperative chylothorax for lung cancer and esophageal cancer]. AB - OBJECTIVE: To define the clinical features of postoperative chylothorax for lung cancer (PCLC), and to compare them with those for esophageal cancer (PCEC). METHOD: We retrospectively analysed clinical characteristics of 12 patients with chylothorax among 4 084 patients receiving resection of lung cancer, as well as 52 in 4 479 patients having resection of esophageal cancer since 1985 at our hospital. RESULTS: The incidence of PCLC was 0.29% and that of PCEC was 1.16%. The percentage of diagnosis confirmed within 4 postoperative days was 33.3% for PCLC, and 76.9% for PCEC. The rate of typical chylous pleural effusion was 83.3% for PCLC, and 5.8% for PCEC. Symptoms and signs of PCLC were much milder than those of PCEC. The re-operation rate was 16.7% for PCLC, and 96.2% for PCEC. All patients were discharged uneventfully. CONCLUSION: The incidence, causes, clinical manifestations, diagnosis, and treatment of PCLC is different from those of PCEC. PMID- 12760760 TI - [Surgical treatment of acute embolism of the upper extremity]. AB - OBJECTIVE: To study the methods of surgical treatment and the prognosis of acute embolism of the upper extremity. METHODS: Balloon catheter embolectomy through the brachial artery was performed in 18 patients with acute embolism of the upper extremity. RESULTS: Both the pulse of the radial and ulnar artery could be palpated in 8 patients, either the pulse of the radial or ulner artery could be palpated in 9 patients. The temperature of the upper extremity was increased in the patient whose embolectomy was performed in the 6th day after onset of the illness. Three patients died postoperatively. CONCLUSIONS: Embolectomy through the brachial artery is an effective method to treat acute embolism of the upper extremity. Elderly and heart and pulmonary diseases are the high risk factors for postoperative death. PMID- 12760761 TI - [Relationship of survival rate, hematoma thickness and midline shift in patients with acute subdural hematomas]. AB - OBJECTIVE: To estimate outcomes of patients with acute subdural hematomas by analysing the hematoma thickness, midline shift and the differences between them. METHODS: Ninety-five patients with acute subdural hematoma were retrospectively studied by calculating hematoma thickness, midline shift and their difference with a statistical analysis of Kaplan-Meier, Wilcoxon-Mann-Whitney U test. RESULTS: The hematoma thickness ranged from 5.0 to 40.0 mm and midline shift was from 0 to 35.0 mm. Among these patients, 51% died and 49% survived after surgery. 18 patients (19%) showed good or satisfactory results. Kaplan-Meier analysis proved that the survival for patients with hematoma thickness approximately equal to l7 mm and a midline shift 15 mm or whose midline shift exceeded hematoma thickness by 2.2 mm, the survival rate was 50%. Glasgow outcome scale scores were correlated significantly with these parameters. CONCLUSION: The hematoma thickness, midline shift and their difference provided a database from which criteria could be derived, that is crucial for prognosis estimation. PMID- 12760762 TI - [Characteristics on diagnosis and treatment of renal tuberculosis]. AB - OBJECTIVE: To study the new characteristics on diagnosis and treatment of renal tuberculosis (RT). METHODS: Eighty-seven patients with renal tuberculosis were retrospectively reviewed; their diagnosis was established by standard microbiological and histological techniques. RESULTS: Atypical RT was diagnosed by various examinations, including urinary analysis, polymerase chain reaction of tuberculosis (PCR-TB), ultrasonography, intravenous urography (i.v.U), and computerized tomography (CT). Treatment consisted of antituberculous chemotherapy in all patients, in combination with nephrectomy (62.5%) or enterocystoplasty (4.6%). CONCLUSIONS: The differential diagnosis of RT should be emphasized, especially for atypical RT, provided a much more specific diagnosis in clinical suspicion of RT. i.v.U can not be regarded as a specific examination for RT. Computerised tomography (CT) can be used for early diagnosis of RT. Surgery for RT is still ablative. PMID- 12760763 TI - [Experimental study of total nutrient admixture promoting transfection of plasmid DNA mediated with liposomes to colorectal cancer cells]. AB - OBJECTIVE: To study total nutrient admixture (TNA) promoting plasmid DNA transfection mediated with liposomes to colorectal cancer cells. METHODS: Dispensing varied transfection agents of liposome + DNA plasmid pEGFP-N(1), TNA + liposomes + pEGFP-N(1), TNA + pEGFP-N(1), liposomes merely, and TAN sole. Human colorectal cancer cell LoVo and HR-8348 were treated with the agents respectively. Green fluorescence protein (GFP) gene as a report gene was detected. RESULTS: GFP was not detected in cancer cells treated with agents of merely liposomes and TAN. Transfection rates of GFP in two groups of cancer cells treated with agent of TNA + liposomes + pEGFP-N(1) were 33%, 38% respectively. With liposome + pEGFP-N(1), the rates of transfection in two cells were 22%, 24% respectively. The expression of GFP was 1% in the two groups of tumor cells treated with TNA + pEGFP-N(1). With agent of TNA + liposomes + pEGFP-N(1), a high transfection rate of GFP gene was obtained. And no negative effect was observed to stabilization of TAN solution. CONCLUSION: TNA may enhance transfection rate of plasmid DNA mediated with liposome, and may be beneficial to the treatment of cancer. PMID- 12760764 TI - [Acellular nerve allograft by chemical extraction in humans]. AB - OBJECTIVE: To develop a procedure by which Schwann cells and myelin in the peripheral nerve could be removed while the basal lamina tubes remained intact, and to obtain a thick and long acellular nerve allograft in humans. METHODS: Four ulnar nerves 10.0 cm long and 4.0 - 5.0 mm in diameter were excised from a donated male body and cleaned from external debris. The nerves were treated with a solution of Triton X-100 and a solution of sodium deoxycholate at room temperature. After a final wash in water, the nerves were stored in phosphate buffered saline (PBS, pH 7.2) at 4 degrees C. HE, luxol fast blue and fibrin staining were performed to visualize cells, myelin and basal membranes respectively and immunohistochemical staining was performed to visualize the presence of laminin, a Schwann cell lamina component, both in fresh and acellular nerve segments. To reveal overall structure better, methylene blue-fuchsin staining was performed in semithin section. The ultrastructure of acellular and fresh nerves were observed and photographed in a transmission electron microscope. RESULTS: The acellular human ulnar nerve was white long cylinder with well elasticity and ductility. HE, myelin and fibrin staining revealed that cells, axons and myelin sheath were removed and basal membrane was preserved after extraction procedure. Staining for the presence of laminin showed that the Schwann cell basal lamina component were present in the nerves after chemical treatment. Methylene blue-fuchsin staining and transmission electron microscopy showed that the myelin sheaths were absent in the extracted nerve segments and empty basal lamina tubes remained in the endoneurium. CONCLUSIONS: We developed an extracted procedure with the detergents of Triton X-100 and deoxycholate, by which cells, axons and myelin sheaths could be removed from a human ulnar nerve while the basal lamina tubes remain intact and a thick long acellular nerve allograft is obtained. The laminin, a Schwann cell basal lamina component, can be preserved in the acellular nerve. PMID- 12760765 TI - [In vivo and in vitro evaluation of inflow cannula of left ventricular assist]. AB - OBJECTIVE: To develop an inflow cannula of left ventricular assist implanted by blood vessel. METHODS: The maximum inflow and properties against folding of 8 sorts of cannulae were measured in mimic extracorporeal circulation appliances and canines. RESULTS: The maximum flow of the cannula increased, as the inner diameter became greater (P < 0.01) compared with each group. The maximum flow rate was (1.82 +/- 0.03) L/min, (2.44 +/- 0.03) L/min, (3.02 +/- 0.04) L/min, (3.31 +/- 0.03) L/min respectively for polyvinyl cannulae with wall thickness of 0.5 mm (PV 0.5 cannula) and inner diameter of 3 mm, 4 mm, 5 mm, 6 mm; (1.83 +/- 0.03) L/min, (3.07 +/- 0.04) L/min respectively for the polyvinyl chloride cannula with wall thickness of 1.0 mm imbedded by spring wire (PVCSW 1.0) and inner diameter of 3 mm and 5 mm; (1.82 +/- 0.02) L/min, 1.84 +/- 0.02 L/min for strengthened polyvinyl cannula with wall thickness of 0.8 mm (SPV 0.8) and inner diameter of 3 mm and polyvinyl cannula with wall thickness of 1.0 mm (PV 1.0 cannula) of inner diameter of 3 mm. There was no remarked statistical difference in vitro maximum flow among the four cannulae of 3 mm inner diameter in vitro. PVCSW 1.0 was showed the best antifolding property, PV 1.0 cannula good and SPV 0.8 and PV 0.5 unsatisfactory in properties against fold. There was no significant statistical difference between in vivo and in vitro maximum flow for PVCSW 1.0 and PV 1.0 cannulae of 3 mm inner diameter. But for SPV 0.8 and PV 0.5 cannulae of 3 mm inner diameter, there was a significant difference between in vivo and in vitro. CONCLUSIONS: PV 0.5 cannula and SPV 0.8 cannula are not suitable to clinical use. PV 1.0 cannula can be used in clinics. PVCSW 1.0 cannula is fully qualified for inflow conduit of left ventricular assist in surgery. PMID- 12760766 TI - [Relevant factors and management for patients with squamous carcinoma in tongue without clinical cervical lymphatic node metastasis]. AB - OBJECTIVE: To discuss the treatment of cN(0) tongue carcinoma patients. METHODS: 185 cases of the mobile tongue carcinoma patients (male 102, female 83, aged 28 to 88) treated with surgery from 1988.5 to 1995.6 had been followed up and retrospectively analyzed. Extensive resection of the primary tumors and neck dissections were performed, and all the samples were pathological positive. RESULTS: The cervical lymphatic node metastasis rates for stage I-II, III-IV disease, grade I, II disease were 16.66%, 38.05%, 17.42% and 37.50% respectively. And the rates were 9.00%, 31.37% and 55.55% for submucous infiltration, muscle infiltration and perineural infiltration, respectively. The overall 5 year survival was 72.43%, and the 5 year specific survival rate was 44.44% and 83.96% for those having or not having cervical node metastasis. The levels of 29 patients with positive node metastasis for 148 cN(0) patients were submandibular and submental lymphatic nodes (22.64%), superior deep cervical lymphatic nodes (35.84%), middle deep cervical lymphatic nodes (26.41%), inferior deep cervical lymphatic nodes (15.09%), posterior neck lymphatic nodes (0.00%). The over all 5 year survival rates for selective neck dissection were 85.13% and 21.62% in therapeutic dissection (chi(2) = 29.73, P < 0.01). Patients performed selective neck dissection the rates were 68.96% and 89.07% respectively with or without lymphatic node metastasis. Comparably the 5-year rate was only 20.00% for the patients performed the therapeutic dissection with lymphatic node metastasis. CONCLUSIONS: (1) cN(0) patients should be observed carefully in stage I, and the selective dissection must be performed in stage II-IV. (2) Supraomohyoid ND is essential for T(2) patients, and functional ND is essential for T(3 - 4) patients. (3) There is correlation between cervical metastasis and the stage, grade or infiltration of tongue cancer (P < 0.05). The prognosis could be expected from these factors. PMID- 12760767 TI - [Functional implantation reconstruction of acquired jaw defects after tumor resection]. AB - OBJECTIVE: Partial or full defects of jawbone following tumor resection frequently lead to a loss of mastication, an impaired speech function and a severe deformity of appearance. To improve the life quality of such patients, implantation or bone grafting-combined implantation was utilized to functionally reconstruct the jaw defects. METHODS: 1. Fragmental or full mandibular defects were reconstructed by vascularized or non-vascularized bone grafts in combination with immediate or delayed implants. 2. The unilateral maxillary defect with an edentulous counter-side was rehabilitated by a prosthesis secured on implants placed on the healthy side. 3. For bilateral maxillary defect, implants were placed in the zygmatic bone or augmented zygmatic bone to support a prosthesis with magnetic retention. RESULTS: 64 jaw defects (10 in maxilla; 54 in mandible) were reconstructed by three different methods to restore the appearance and functions. The longest follow-up period was 12 years and the shortest 5 year, only 6 implants were lost due to failure of osseointegration. The implant survival rate for the maxillary defects was 97.5%, for the mandibular defects with vascularized bone grafts was 97.1%, and with non-vascularized bone grafts was 97.7%. CONCLUSIONS: Implants-borne prosthesis is an applicable technique in restoration of maxillary defects. In case of insufficient zygomatic thickness, bone augmentation is often needed prior to implantation. As for the mandibular reconstruction, bone grafting in combination with implantation is an ideal method. Compared to non-vascularized bone grafting, the vascularized method is much more suitable for bone grafting beds with poor blood supply. From the practical point of view, non-vascularized bone graft in combination with implantation is more practicable. The reduced off-body time of bone grafts from donor to recipient site keeps most of osteoblasts vital and enables simultaneous implants to achieve osseointegration. This confirms the osteogenesis, osteoconduction and osteoinduction of bone autografts. PMID- 12760768 TI - [Surgical anatomy and preservation of the accessory nerve in radical functional neck dissection]. AB - OBJECTIVE: The surgical anatomy and preservation of the accessory nerve in radical functional neck dissection were studied. METHODS: Thirty-three cN(0) patients with oral cancers were entered into the study. Radical functional neck dissection were performed and the relations between the accessory nerve and its surrounding structures were recorded. RESULTS: The accessory nerve going through or beneath the sternocleidomastoid muscle occurred in 82% (27/33) and 18% (6/33) of the patients respectively. Communicating branches between the accessory and the cervical nerves were found in 85% (28/33). There was 2 to 3 cm of the accessory nerve paralleled to the anterior border of the trapezius muscle before it entered the muscle in 70% (23/33). The dissection of the nerve needed 20 to 30 minutes. Twenty-seven percent of the patients had pathologically proved lymph node metastases. CONCLUSIONS: Looking for accessory nerve under the upper portion of the sternocleidomastoid muscle and above the middle point of the muscle posterior border is simple and safe. The point of the great auricular nerve going out the muscle is an important indicator for finding the accessory nerve. PMID- 12760769 TI - [Transfection of the nm23-H1 gene into BcaCD885 cell line inhibits the potential of invasion, adhesion and mobility]. AB - OBJECTIVE: To transfect nm23-H1 into the BcaCD885 cell lines in order to get safe high-efficiency and low-toxicity, and to find out whether nm23-H1 could affect the invasion and metastases ability of BcaCD885 cell lines. METHODS: Lipofect was used to transfect nm23-H1 into BcaCD885 cell lines; immunohistochemistry was used to detect the difference expression of nm23-H1 between transfected and non transfected cell lines; then transwell-room and wash way were used to detect the difference of invasion and metastases ability between transfected and non transfected cell lines. RESULTS: PCMV-NEO-BAM system gave the stability expression of nm23-H1; there was significant different NDPKA expression between transfected and non-transfected BcaCD885 cell lines; the invasion and metastases ability of transfected BcaCD885 cell lines decreased obviously. CONCLUSION: nm23 H1 can inhibit the metastases of BcaCD885 cell lines significantly. PMID- 12760770 TI - [Investigation of apoptosis mechanism of arsenic trioxide on oral squamous cell carcinoma]. AB - OBJECTIVE: To probe the possible mechanism of growth-inhibitory and apoptosis of oral squamous cell carcinoma by arsenic trioxide. METHODS: The induction of apoptosis in two tongue squamous carcinoma cells treated by arsenic trioxide was investigated. The morphology changes of the cells was observed under electron microscope. The mitochondrial transmembrane potential was detected using rhodamine 123 and flow cytometry. The cell cycle was detected by flow cytometry, and p16, p53, BCL-2, Caspase-3, and PARP changes were examined by western blot. RESULTS: 1. The antiproliferative effect on the oral squamous carcinoma cells by arsenic trioxide was carried out through two ways: induction of apoptosis and toxicity damage. 2. Activation of the caspase-3 and PARP, while no changes of p16, p53, BCL-2 occurred. 3. Mitochondrial transmembrane potential collapse and G(2)-M stagnation were correlated with apoptosis of oral squamous cell carcinoma. CONCLUSIONS: 1. Tubulins and mitochondria may be the chief action position of arsenic trioxide, which is the start-up factors of mechanism. 2. Activation of the caspase-3 proteolytic pathway may be one of the pivotal ways of apoptosis procedure induced by arsenic trioxide. PMID- 12760771 TI - [All-trans retinoic acid augments the bystander effect of herpes simplex virus thymidine kinase/ganciclovir system in the treatment of tongue carcinoma cell line]. AB - OBJECTIVE: To investigate the augmentation effect and mechanism of all-trans retinoic acid (ATRA) on the bystander effect of herpes simplex virus thymidine kinase (HSV-TK)/ganciclovir (GCV) system in the treatment of tongue carcinoma cell line. METHODS: Immunocytochemistry and flow cytometry (FCM) were used to analyze the expression of Cx43 protein in Tca8113 cells after treated with ATRA; The bystander effect of HSV-TK/GCV system on tongue carcinoma cells before and after treatment with ATRA was detected by MTT assays. The interaction of ATRA and bystander effect was analyzed by factorial experiment. RESULTS: After treated by ATRA (10(-7) mol/L - 10(-5) mol/L), the expression of Cx43 protein was up regulated in Tca8113 cells and the positive rate of Cx43 protein increased from 5.17% (before treatment) to 30.53% (10(-5) mol/L ATRA). There was significant difference between ATRA treated cells and untreated cells (P < 0.01). The bystander effect of HSV-TK/GCV system on Tca8113 cells was poor, but improved after combined with ATRA. There was cooperation effect between ATRA and bystander effect of HSV-TK/GCV (P < 0.05). CONCLUSIONS: ATRA can augment the bystander effect of HSV-TK/GCV system in the treatment of tongue carcinoma cell line. The mechanism might be due to up-regulation of Cx43 gene and restore gap junction intercellular communication. PMID- 12760772 TI - [Cloning and polymorphism analysis of prtH gene from Porphyromonas gingivalis]. AB - OBJECTIVE: To clone the prtH gene from Porphyromonas gingivalis (P.g) ATCC 33277 and analyze the polymorphism of prtH gene from 5 strains of P.g in order to explore the relationship between P.g and periodontitis. METHODS: Using PCR, the prtH was amplified and cloned into pGEM-T vector. To illustrate the prtH polymorphism among P.g strains, the genomic DNAs were extracted and screened by PCR with three pairs of specific primers, dot blot and Southern blot hybridization using the biotin-labeled prtH sequence as probe. RESULTS: Recombinant DNA pGEM-T- prtH was verified by restriction endonuclease and sequence assay. Strain W 381 and ATCC 33277 showed the identical results in PCR and hybridization assays, whereas strain ATCC 49417 and 14-3-2 revealed individual hybridization patterns. Strain 47A-1 seemed even not to contain prtH gene. CONCLUSIONS: Different prtH gene sequences exist in different P.g strains. This polymorphism may indicate various potential virulent effects during the infection and pathogenesis. Established PCR protocol is sensitive for identification of prtH gene. PMID- 12760773 TI - [Correlation between prognosis and cell proliferation, apoptosis in squamous cell carcinoma of tongue]. AB - OBJECTIVE: To study the correlation between prognosis and cell proliferation, apoptosis in squamous cell carcinoma (SCC) of tongue. METHODS: Using TdT-mediated dUTP biotin nick end labeling (TUNEL) for cell apoptosis and immunohistochemical staining for proliferating cell nuclear antigen (PCNA), Bcl-2 and Bax protein expression. RESULTS: In SCC of tongue, the positive rates of apoptosis index (AI), Bcl-2, Bax and PCNA were 55.8%, 65.4%, 73.1% and 100.0% respectively. In the univariate Kaplan-Meier analysis, low expression of Bax, high expression of PCNA, Bcl-2/Bax expression ratio > 1 and high staging were all associated with lower survival rate (Log-rank test, P < 0.05, respectively), AI was only correlated with grading. Bcl-2 did not show a significant association with all features. CONCLUSIONS: The levels of Bax and PCNA expression and Bcl-2/Bax expression ratio have independent prognosis value in squamous cell carcinoma of the tongue. Comprehensive analysis of these parameters can be helpful to identify biological behavior and predict more accurately the prognosis of patients of tongue squamous cell carcinoma. PMID- 12760774 TI - [The study of natural head posture in edentulous patients before and after oral rehabilitation]. AB - OBJECTIVE: To study the natural head posture (NHP) of edentulous patients compared with dentate people and to understand NHP shift immediately inserting complete dentures and three months after denture wearing. METHODS: The NHPs of twenty edentulous patients and twenty-two dentate subjects were recorded with cephalometric technique. The NTPs of edentulous patients were exposed three times, before inserting, right after inserting and three months after inserting dentures. The NHPs of dentate subjects were recorded once as a controlled group. Six angles measured from the radiographs were evaluated for the NHP and three angles and two linear measures for the mandibular position. RESULTS: Compared with dentate subjects, the craniomandibular angles of the edentulous patients increased and the camber of the cervical columns decreased, but changes of the NHP were not significant. The mandible showed a significant upward moving (P < 0.05) and an insignificant forward moving. At the time of denture insertion, the camber of the cervical columns significantly increased (P < 0.05). Three months after using the dentures, the craniomandibular angles of the patients obviously decreased. (P < 0.05). CONCLUSION: The shift of NHP after denture inserting does exist. The shifted NHP of the patients with dentures is a new balanced posture and closer to the normal position than the NHP of them without dentures. PMID- 12760775 TI - [The roles of Smad 2/3 in transforming growth factor-beta 1 signaling in human dental pulp cells]. AB - OBJECTIVE: To explore the roles of Smad 2/3 in transforming growth factor-beta(1) (TGF-beta(1)) signaling by human dental pulp cells. METHODS: Laser scanning confocal microscope was used to observe translocation of Smad 2/3 from plasma into nucleus in cultured dental pulp cells at early stage of TGF-beta(1) treatment, and changes of Smad 2/3 protein expression at later stage were evaluated by Western blot analyses. RESULTS: The expression of Smad 2/3 (fluorescence intensity) kept decreasing in cytoplasm but increasing in nucleus within 2 h after TGF-beta(1) treatment, forming a trend that Smad 2/3 translocated into nucleus from cytoplasma. The total amount of Smad 2 protein remained unchanged before and after TGF-beta(1) treatment, but the expression level of Smad 3 decreased markedly after 24 h treatment and kept dropping by 48 h. CONCLUSIONS: The results suggest that the Smad 2/3 may be the downstream signal transducers of TGF-beta(1) in human dental pulp cells and Smad 2/3 may mediate TGF-beta(1) signaling by translocation early in TGF-beta(1) treatment, while down-regulation of Smad 3 expression by TGF-beta(1) at later stage is involved in negative modulation of TGF-beta(1) signaling. PMID- 12760776 TI - [Orthodontic treatment of malocclusion with mandibular congenital missing teeth guided by Bolton index]. AB - OBJECTIVE: To evaluate the outcome of the treatment of malocclusion with mandibular congenital missing teeth guided by Bolton index. METHODS: 26 class I and II cases (males 10, females 16) were chosen in this study. Guided by Bolton index, enamel stripping, denture and two types of tooth extraction were used in the treatment. RESULTS: Normal Bolton index was achieved and the results of treatment were satisfactory. CONCLUSIONS: Malocclusion with mandibular congenital missing teeth can be treated successfully guided by Bolton index. PMID- 12760777 TI - [Study on retention and stability of linear occlusal complete dentures]. AB - OBJECTIVE: To learn retention and stability of linear occlusal complete dentures by investigating the subjective feelings of patient and the value of retention force. METHODS: Static retention forces of maxillary and mandibular dentures were measured for 25 patients wearing linear occlusal dentures by using Hz-1 retention dynamometer. The subjective feelings of patients in functional state were gained simultaneously through questionnaire. RESULTS: Linear occlusal dentures demonstrate good retention in static and dynamic state. Among patients with severe resorption of residual ridge (RRR), mandibular linear occlusal dentures (shown good retentive subjective feelings) demonstrate significantly smaller retention force than those with slight or medium degree of RRR. There is no correlation between the subjective feelings and the values of retention forces of mandibular dentures. The subjective feelings of patients wearing new linear occlusal dentures are much better than that of old anatomic occlusal dentures. CONCLUSION: Linear occlusal dentures improve the performances of dentures by enhancing their stability during mastication movement. PMID- 12760778 TI - [Orthokeratinized odontogenic cyst: a clinicopathological and immunocytochemical study]. AB - OBJECTIVE: To investigate the clinicopathological features and behavior of a group of jaw cysts with a solely orthokeratinized lining epithelium. METHODS: 20 cases of this cyst type were reported under the term of orthokeratinized odontogenic cyst (OOC) and their clinical, histological and immunocytochemical features were compared with that of odontogenic keratocyst (OKC). RESULTS: The cysts of the present series were all solitary lesions, occurred mostly in young male patients, and showed a predilection for the posterior mandible areas. Follow up of 15 patients revealed no recurrence following enucleation. Histological and immunocytochemical studies indicated that OOC epithelium lacked the typical features of OKC and appeared to show a lower proliferative activity. CONCLUSION: These findings suggest that OOC is clinicopathologically distinctive from OKC and may thus constitute its own clinical entity. PMID- 12760779 TI - [Phenotypic variation in Actinobacillus actinomycetemcomitans]. AB - OBJECTIVE: To investigate the colony variation in Actinobacillus actinomycetemcomitans (Aa) from rough to smooth and recognize its different morphology during laboratory translations. METHODS: Primary strains isolated from subgingival plaque of two juvenile periodontitis patients were repeatedly subcultured on agar plates and broth; for broth culture, every generation was translated in broth and on solid medium separately to observe the corresponding morphologies of Aa grow in broth. RESULTS: Three smooth strains of Aa from the broth culture were obtained. The process was about 7-8 generations: colonies changed from a small and adherence phenotype to a bigger and sediment ones and finally the culture supernatant became turbid; the corresponding morphologies grow on agar exhibiting an adherent, small rough colony phenotype which had a star-shaped internal structure converted gradually to a kind of bigger, opaque, nonadherent, smooth phenotype, then the colony extended out from the margin of the colony and finally converted to a flat, almost parent morphology and the same time the star-like inner structure converted to a simpler and smaller type and finally disappeared. We could not get completely smooth variants of Aa from agar. CONCLUSIONS: The variation in colony morphology of Aa from rough to smooth is a process, in which the colony was gradually wetter and bigger and at the same time gradually lost the inner structure. During this process three colony morphologies at least can be seen, including rough, opaque smooth and almost translucent smooth. PMID- 12760780 TI - [The effect of dentin phosphoprotein on inducing mineralization]. AB - OBJECTIVE: To investigate the effect of dentin phosphoprotein (DPP) in inducing dentinal mineralization. METHODS: Human DPP was combined with EAH-Sepharose 4B beads and its function of inducing mineralization was studied in mineralization system in-vitro. The mineral formed on the surface of the beads was analyzed by scanning electron microscopy (SEM) and the structure was analyzed by X-ray diffraction and plasma emission spectrum. RESULTS: There was mineral formed on the beads with combined DPP and the mineral was calcium phosphates whose ratio of calcium to phosphate was 1.33. The diffractogram of the formed mineral was more similar to hydroxyapatite than to other calcium phosphates. CONCLUSION: When tightly combined with certain support substance, human DPP can induce mineralization. PMID- 12760781 TI - [In vivo study of remineralization with trace elements by laser scanning confocal microscope]. AB - OBJECTIVE: To evaluate in vivo the remineralization containing trace elements. METHODS: The volunteers were selected by pre-designed criteria of adopting and eliminating. Caries-like lesions were prepared in the enamel of extracted human premolars with the use of demineralizing solution. Sections of the normal and lesion enamel (approximately 2 mm x 2 mm) were prepared, with the cut surfaces protected by nail varnish. 2 enamel specimens were mounted in a removable appliance. By measuring lesion parameters (area, total and average dye fluorescence) on a tooth with confocal laser scanning microscopy (CLSM), the effect of remineralization was assessed. Specimens were cut and stained with a fluorescent dye (0.1 mmol/L rhodamine B) for 1 h and analyzed using CLSM. RESULTS: CLSM detected significantly greater remineralization (P < 0.05) in the specimens treated with the trace elements fluoride-containing solution and only containing fluoride (P < 0.05), especially more obvious difference was shown upon remineralized solution with trace elements, which means it produced a greater remineralization. CLSM data of remineralized solution with trace elements were showed: delta Z vs. Area = -50.4 +/- 8.1; delta Z vs. TF = -27.8 +/- 3.8; delta Z vs. AF = -91.5 +/- 8.9. CONCLUSIONS: The ability of remineralization of the new solution is better than that only containing fluoride in in vivo study. It can potentially prevent initiation of caries. This study provides not only the theoretical foundation for clinical application, but also shows a new kind of experimental method in the study of demineralization and remineralization. PMID- 12760783 TI - [Influence of super high molecular weight poly D,L-lactic acid on viability and new bone formation of osteoblasts]. AB - OBJECTIVE: To investigate the influence of the viability and new bone formation of osteoblasts by the super high molecular weight poly D,L-lactic acid (SHMW PDLLA). METHODS: 1. The osteoblasts derived from neonatal rat were grown and maintained at steep of SHMW-PDLLA and normal culture medium. The viability and function of the osteoblasts were measured with MTT array. 2. The plate and screws made of SHMW-PDLLA were implanted and fixed at the artificial fractured mandible of dogs. Specimens were gained at 3 and 6 months and examined with macroscopy and SEM. RESULTS: 1. There is no significant difference of OD values between the experimental group and the control group (P > 0.05). The SHMW-PDLLA isn't toxic to osteoblast at 1 week and 2 weeks, and the toxicity is 3% at 3 days. 2. There were a lot of new bone formed between the implanted SHMW-PDLLA plate and bone tissues under SEM. CONCLUSION: SHMW-PDLLA hasn't pathological influence on the viability and new bone formation of osteoblasts and it is feasible in tissue engineering of bone. PMID- 12760782 TI - [Experimental study of the isolation, culture and in chondrogenic differentiation of human bone mesenchymal stem cell]. AB - OBJECTIVE: To study the isolation of human bone marrow mesenchymal stem cells (MSCs) and in vitro differentiation into chondrocytes as potential seed cell for condyle cartilage tissue engineering. METHODS: Human MSCs were isolated by percoll solution from normal human bone marrow sample and cultured in flasks. Specific cell surface markers were identified by flow-cytometry. After the cells were treated with inductive medium containing insulin, transferrin, pyruvate, dexathemesone and TGF-beta for 7 - 14 days, microscopic, histological and immuno histo-chemical studies were performed for chondrogenic phenotype identification. RESULTS: Primary cultures of human MSCs express CD29 and CD44 positively and meanly, but CD34, CD45 and HLA-DR negatively. After 14 days of induction, the cells were positively stained by safranin O. Immunohistochemical analysis proved strong type II collagen expression. CONCLUSIONS: Percoll helps to generate a better isolation of MSCs from human bone marrow aspirates with a purity more above 95%. The isolated MSCs can be expanded and induced in vitro to differentiate into chondrocytes by inductive medium. PMID- 12760784 TI - [The in vivo formation of cementum-like tissue by bovine cementoblasts]. AB - OBJECTIVE: To test the bovine cementoblasts (CBs) cementum-forming ability in vivo. METHODS: Root fragments of newborn bovine freshly extracted mandibular incisor were cultured routinely and 4th-5th passages of CBs were harvested. CBs were then cultured in the medium supplemented with 50 mg/L alpha-ascorbic acid and 10 mmol/l beta-glycerolphosphate to form a thick layer as tissue engineering scaffold for cementum formation. Collagen membrane was used as control scaffold. 2 x 10(6) cells were attached to the CBs-made carrier as well as collagen membrane scaffolds and transplanted subcutaneously into immunodeficient mice. Transplants were harvested at 7th week. Histological sections were stained with HE, alizarin red S and van Kossa methods as well as monoclonal Ab against bovine cementum attachment protein (CAP). RESULTS: CBs-made scaffold supported more cementum-like tissue (CLT) formation than collagen-made scaffold. The CLT formed on CBs scaffold was partly calcified with embedded cells. Uncalcified cementoid like material could be seen on the surface and was encircled by cubical CB-like cells. The CLT was also positive to CAP and van Kossa staining. CONCLUSIONS: These results suggest that the bovine CBs can form cementum-like tissue. The cell made carrier is a better scaffold than collagen membrane. PMID- 12760785 TI - [A study on absorption and utilization of calcium, iron and zinc in mineral fortified and dephytinized soy milk powder consumed by boys aged 12 to 14 years]. AB - OBJECTIVES: In order to understand the absorption and utilization of calcium, iron and zinc in cow milk, soy milk powder and dephytinized soy milk powder in the children and adolescents, and to lay a foundation for the improvement of protein and mineral nutrition in Chinese residents with popularizing consumption of soy milk powder. METHODS: Totally, 57 boys aged 12 to 14 years were selected and divided into three groups, matched by age, hemoglobin concentration, height and weight. The boys were given 220 ml of fortified and dephytinized soy milk, soy milk or cow milk, respectively, containing calcium 270 mg, iron 4 mg and zinc 4 mg, which were labeled with stable isotopes (44)Ca, (58)Fe and (70)Zn in a single serving size, and their feces were labeled with brilliant blue and dysprosium. Feces specimens were collected for all the subjects and absorption rate of calcium, iron and zinc were measured for all the children who took fortified and dephytinized soy milk, soy milk or cow milk, respectively. Calcium in feces was measured with heat ionized mass spectrometry, and recovery of (58)Fe and (70)Zn in feces and content of dysprosium were measured with induction coupling iso-ionic mass spectrometry, and then absorption rates of iron and zinc were calculated and adjusted by the recovery rate of dysprosium. RESULTS: Iron absorption rate was (6.7 +/- 3.8)% in soy milk powder group, (15.5 +/- 9.2)% in the cow milk group and (20.6 +/- 7.3)% in dephytinized soy milk powder group, respectively. Calcium absorption rate was (43.5 +/- 10.7)%, (64.2 +/- 11.4)%, and (50.9 +/- 6.6)% in the three groups, respectively. Absorption rate of zinc was (11.3 +/- 6.5)%, (31.2 +/- 10.4)% and (20.1 +/- 7.4)%, respectively. Significant increase in absorption of calcium, iron and zinc was observed in the groups with fortified and dephytinized soy milk powder, as compared with those consuming nondephytinized soy milk powder. Absorption rate of calcium and zinc was significantly higher in the group with cow milk than that in the group with dephytinized and fortified soy milk powder, and iron bioavailability was lower in the group with cow milk than that with dephytinized soy milk powder, with no significant difference. Absorption rates of calcium, iron and zinc were higher in children of China due to their long term adaptation to dietary intake of lower mineral and protein, and higher fiber. CONCLUSIONS: Bioavailability of calcium, iron and zinc in soy milk powder could be increased by dephytinized treatment for it. PMID- 12760786 TI - [Effect of long term supplementation of mineral-fortified dephytinized soy milk powder on biomarkers of bone turnover in boys aged 12 to 14 years]. AB - OBJECTIVES: In order to understand the effect of long term consumption of mineral fortified dephytinized soy milk powder on skeletal health in children, biological effects of mineral-fortified dephytinized soy milk powder, dephytinized mineral unfortified soy milk powder and fresh mineral-fortified cow milk were compared. METHODS: Totally, 90 boys aged 12 to 14 years were involved in the study and were given daily with mineral-fortified dephytinized soy milk powder, dephytinized soy milk powder and fresh fortified cow milk, respectively, for two months. The blood samples were collected from all the subjects for measuring activity of serum bone specific alkaline phosphatase and serum level of oesteocalcin, and the urine samples were collected for determining urine C-terminal telopeptides of type I collagen (CTx). RESULTS: After two-month intervention, serum level of oesteocalcin increased significantly in the all groups, (16.42 +/- 11.25) micro g/L for the group with fortified soy milk powder, (18.86 +/- 11.57) microg/L for the group with unfortified soy milk powder and (13.81 +/- 10.06) microg/L for the group with fortified cow milk, respectively. The activity of bone specific alkaline phosphatase significantly decreased in the all groups, by (0.28 +/- 0.40) micromol x s(-1) x L(-1) for the group with fortified soy milk powder, (0.26 +/- 0.35) micromol x s(-1) x L(-1) for the group with unfortified soy milk powder, and (0.17 +/- 0.28) micromol x s(-1) x L(-1) for the group with fortified cow milk, respectively. As well as, urine level of CTx decreased significantly, by (349.1 +/- 380.3) mg/L for the group with fortified soy milk powder, (260.4 +/ 412.4) mg/L for the group with unfortified soy milk powder, and (354.4 +/- 404.7) mg/L for the group with fortified cow milk, respectively. CONCLUSIONS: Long term consumption of mineral-fortified and dephytinized soy milk powder could improve bone mass retention and calcium nutrition status efficiently in children. PMID- 12760787 TI - [Effects of calcium and vitamin D-fortified milk on physical development in school girls aged 10 to 12 years]. AB - OBJECTIVES: To investigate the effects of calcium and vitamin D supplementation in fortified milk on physical development in Chinese school girls aged 10 - 12 years. METHODS: A non-randomized, double-blind intervention trial was carried out at nine primary schools of Beijing for two years. Seven hundred and fifty-seven girls (with an average age of 10.1 years) were recruited from nine primary schools in Beijing, China. They were divided into three trial groups, a control group (n = 259) having regular diet without milk supplementation, Group 1 (n = 238) having regular diet supplemented with 144 ml/d of calcium-fortified milk, and Group 2 (n = 260) having regular diet supplemented with 144 ml/d of calcium and vitamin D-fortified milk. Data of dietary intake and physical activities were collected by questionnaire, and height, weight, sitting height and knee height were measured before, during and after intervention. RESULTS: Two years after intervention, the percentage increases in height and weight were significantly greater in Groups 1 and 2 than those in control group (9.52%, 9.26% and 8.59% in height, and 34.53%, 35.38% and 30.91% in weight, respectively). The percentage increase of sitting height was greater in Group 1 than those in Group 2, and then than that in control group (9.21%, 8.58% and 7.87%, respectively). The percentage increase of knee height was significantly greater in Group 1 than that in Group 2 and control group (9.00%, 8.37% and 8.44%, respectively). CONCLUSION: Calcium and vitamin D-fortified milk supplementation could promote their physical development in girls aged 10 - 12 years. PMID- 12760788 TI - [Roles of leptin in the development during girls'puberty]. AB - OBJECTIVES: To study the roles of leptin in the development during puberty in girls and the its relationship with insulin (INS), growth hormone (GH), estradiol (E(2)) and testosterone (T). METHODS: One hundred and fifty girls with simple obese aged 7 to 17 years, and 150 normal healthy girls and 150 girls with malnutrition matched for age (+/- 3 months) and height (+/- 2 cm) were selected. Serum levels of leptin, INS, GH, E(2) and T were measured for them. RESULTS: Their serum level of leptin positively correlated with body mass index (BMI) and age. Serum level of leptin in girls increased steadily from Tanner stage B(1) to stage B(5). At Tanner stage B(2), serum level of leptin in the normal groups (7.72 microg/L) was not significantly different from that in those with malnutrition (7.36 microg/L), but significantly lower than that in the obese groups (12.85 microg/L). At other Tanner stages, there was significant difference in serum level of leptin among obese, normal and malnutrition groups. Serum level of leptin correlated negatively with serum GH and positively with serum INS, but not correlated with E(2) and T. CONCLUSIONS: Leptin may play a role in triggering development during puberty in girls. Serum level of leptin at Tanner stage B(2) may be the threshold dose to trigger the onset of puberty in girls. Quickly increasing level of leptin at Tanner stage B(5) may inhibit the increase of GH, which ushered the end of puberty in girls. PMID- 12760789 TI - [Relationship of the hair content of rare earth elements in young children aged 0 to 3 years to that in their mothers living in a rare earth mining area of Jiangxi]. AB - OBJECTIVE: To study the relationship of hair content of rare earth elements (REEs) in the young children aged 0 - 3 years to that in their mothers living in a rare earth mining area of Jiangxi Province. METHODS: Content of five kinds of REEs, i.e., lanthanum (La), cerium (Ce), praseodymium (Pr), neodymium (Nd) and samarium (Sm) were determined for the hair-root samples collected from 71 young children and 62 of their mothers by inductively coupled plasma source mass spectrometry (ICP-MS). RESULTS: The mean hair content of REEs (e.g., La) was the highest (2,202.90 ng/g) in the young children living in the place nearest to the REE mining area, next (471.72 ng/g) was in those nearer to the REE mining area, and the lowest (97.37 ng/g) was in those in the control area. And, the hair content of REEs in the mothers was the same as that in their children, i.e, the mean content of La was the highest (1,510.21 ng/g) in high-exposure area, next (241.63 ng/g) was that in the low-exposure area, and the lowest (59.15 ng/g) was that in the control area, with P < 0.001. Pair-comparison analysis for the means showed that the average hair level of five kinds of REEs in the young children was two times high as their mothers' (P < 0.001). For example, the hair level of La was 533.83 ng/g in the young children and 279.78 ng/g in their mothers, respectively. Simple linear regression analysis showed that there was a significant correlation between the hair levels of each kind of REEs in the young children and those in their mothers, with a correlation coefficient of more than 0.75, e.g., that of La was 0.878 (P < 0.01). CONCLUSIONS: The hair level of REEs can be used as a bio-marker to reflect body's level of exposure to REEs. The hair level of REEs in young children and their mothers decreased with the increase of the distance from their home to rare earth mining area. Young children living in the area with REEs mining may be the high-exposure population, and their hair level of REEs was significantly higher than that in their mothers and more attention should be paid to them. PMID- 12760790 TI - [Effects of microcystins on cell cycle and expressions of c-fos and c-jun]. AB - OBJECTIVES: To investigate the effects of microcystins on cell cycle and expressions of c-fos and c-jun, and explore the potential carcinogenic mechanisms of Microcystins. METHODS: Microcystic cyanobacteria extraction (MCE) purified by Sep-Pak C(18) cartridge was added into the media and co-incubated with SHE cell for various periods. Immunohistochemistry assay was applied to detect the expressions of c-fos and c-jun at 1, 3, 6 hr time point, and cell cycle at 6, 12, 24 hr point were analyzed by flow cytometry respectively. RESULTS: Sustained up regulated expression of c-fos and c-jun were induced by MCE during the experimental period, and 5 - 6 folds increased expression were observed at 6 hour point after treatment. As much as 44.8 per cent of cells were induced to entry S phase from resting G(0)/G(1). CONCLUSION: Up-regulating the expression of transcript factor such as c-fos and c-jun thus to induce cell abnormal proliferation may be the potential carcinogenic mechanisms of microcystins. PMID- 12760791 TI - [The effects of conjugated linoleic acid on the expression of invasiveness and metastasis-associated gene of human gastric carcinoma cell line]. AB - OBJECTIVES: To study the effects of c9,t11-conjugated linoleic acid (c9,t11-CLA) on invasive ability of human gastric carcinoma cell line (SGC-7901) and to explore its possible mechanism. METHODS: Reconstituted basement membrane invasion assay was used to evaluate invasive ability of cancer cells. Expression of TIMP 1, TIMP-2 and nm23-H(1) mRNA was measured by reverse transcription polymerase chain reaction (RT-PCR) in SGC-7901 cells. RESULTS: At the concentrations of 200 micromol/L, 100 micromol/L and 50 micromol/L, c9,t11-CLA suppressed their reconstituted basement membrane invasion of SGC-7901 by 53.7%, 40.9% and 29.3%, respectively. c9,t11-CLA could induce the expression of TIMP-1, TIMP-2 and nm23 H(1) mRNA in SGC-7901 cells. CONCLUSIONS: The invasion of SGC-7901 cells could be inhibited by c9,t11-CLA through reconstituted basement membrane. Anti-invasion action of c9,t11-CLA might be associated with induction of expression of TIMP-1, TIMP-2 and nm23-H(1) mRNA in tumor cells. PMID- 12760792 TI - [Cloning, sequencing and expressing of microneme protein 1 partial gene in toxoplasma gondii ZS2 isolate]. AB - OBJECTIVE: To construct a recombinant prokaryotic expression vector (plasmid) containing microneme protein 1 (MIC1) partial gene in toxoplasma gondii (T. gondii) ZS2 isolate. The gene was expressed in varied Escherichia coli (E. coli) after sequencing. METHODS: The gene fragment coding MIC 1 from the genomic DNA of T. gondii ZS2 isolate was amplified by polymerase chain reaction (PCR). The gene was inserted to a prokaryotic expression vector pWR450-1 by digesting with restriction enzymes and linking reaction. The positive clone was screened on LB plates containing ampicillin and identified by restrictive enzyme digestion, PCR amplification and sequence analysis. The recombinant plasmid was transferred into E. coli TG1, JM109 (DE3) and DH5 alpha, and was expressed under the induction of IPTG. The expression products were identified by SDS-PAGE. The MIC1 gene structure was analyzed and compared in homology with the gene sequence of RH isolate using computer software. RESULTS: The recombinant plasmid pWR450-1/MIC1, after cloning from acquired 471 bp MIC1 gene fragment and amplified from the genome gene ZS2, was complete homologous to the sequence of RH isolate, reflecting its highly conservative. The gene could be expressed as fusion protein with 70,000 in varied E. coli. CONCLUSION: Recombinant plasmid pWR450-MIC1 was successfully constructed and could be expressed in different strains of E. coli, laying a foundation for research on its structure and function. PMID- 12760793 TI - [Brain-derived neurotrophic factor mRNA expressions in rat brain induced by short term in vivo exposure to deltamethrin]. AB - OBJECTIVE: To elucidate the mechanism of damage on central nervous system (CNS) caused by deltamethrin (DM). METHODS: The mRNA and protein expressions of brain derived neurotrophic factor (BDNF) in the cerebral cortex and hippocampus of the rats exposed to DM were measured by retro-transcription-polymerase chain reaction (RT-PCR), dot blot, flow cytometry analysis and immunohistochemistry. RESULTS: After exposure to DM at high-dose (DM1, 25.0 mg x kg(-1) x d(-1), i.p.) once and low-dose (DM2, 12.5 mg x kg(-1) x d(-1), i.p.) for 5 days, the level of BDNF mRNA and protein expression in the cerebral cortex and hippocampus of the rats increased significantly. The levels of BDNF mRNA and protein expression in the cerebral cortex and hippocampus measured by of RT-PCR in the rats with DM1 and DM2 were higher than those in the controls by 48% and 56%, and 59% and 54%, respectively. And, those measured by dot blot in the rats with MD1 and MD2 were 186% and 161%, and 148% and 158% of those in the controls, respectively, basically similar to those measured by RT-PCR. Flow cytometric analysis showed that the levels of BDNF mRNA and protein expression in the cerebral cortex and hippocampus in the rats with DM1 and DM2 were higher than those in the controls by 53% and 89%, and 45% and 46%, respectively. Immunohistochemical analysis showed that protein expression in the cerebral cortex of the rats with DM1 and DM2 were 129% and 147% of those in the controls, same as the flow cytometric analysis, but those were significantly higher in the hippocampus mainly in the CA1 and DG areas of the rats with MD1 and the CA3 and DG areas of the rats with DM2. CONCLUSIONS: DM could induce BDNF mRNA and protein expression in the cerebral cortex and hippocampus of the rats, which could play an important role in repairing of nerve damage. PMID- 12760794 TI - [Role of hepatitis B virus infection in pathogenesis of IgA nephropathy]. AB - OBJECTIVE: To clarify the relationship between hepatitis B virus (HBV) infection and IgA nephropathy (IgAN). METHODS: HBV antigen (HBAg) in renal tissues of the patients with IgAN was detected by immunohistochemical technique, the carrier status and localization of HBV DNA in renal tissues were determined by Southern blot analysis and in situ hybridization. RESULTS: Serum HBsAg was detected in 18 of the 100 patients with IgAN (18%), HBAg was detected in 31 of 100 patients (31%) in their renal tissue and in 20 of 31 patients (65%) in their glomeruli, and both HBsAg and HBcAg were detected in 10 of 31 patients (32%), respectively. HBcAg was also found in tubular epithelia (45%, 14/31) and renal interstitium (6%, 2/31), respectively. Five of six cases were proved to be positive of integrated-form HBV DNA in their renal tissue by Southern blot analysis. In situ hybridization demonstrated that HBV DNA was 8/8 and 6/8 positive in their renal tubules and glomeruli of all eight specimens, localized in the nucleus of tubular epithelial cells, glomerular mesangial cells, as well as infiltrated interstitial lymphocytes. CONCLUSION: HBV infection closely related with IgAN and HBV infection might be involved in pathogenesis of IgAN. PMID- 12760795 TI - [A case-control study on risk factors for nosocomial infection by extended spectrum beta-lactamases-producing bacteria]. AB - OBJECTIVE: To explore the risk factors for nosocomial infection caused by extended-spectrum beta-lactamases (ESBLs)-producing bacteria in hospitals of Zhejiang province. METHODS: One hundred and eighty-five cases with nosocomial infection (108 men and 77 women, with an average age of 55 +/- 17 years) caused by positive-ESBLs bacteria, including 59 cases of respiratory infection, 71 with urinary infection, ten with blood infection, 30 with wound infection and 59 with other infection, and 77 controls with nosocomial infection (54 men and 23 women, with an average age of 54 +/- 20 years) caused by negative-ESBLs bacteria, including 38 cases of respiratory infection, 20 with urinary infection, six with blood infection, eight with wound infection and five with other infection, from six hospitals in Zhejiang Province were studied during May 1999 to May 2000. Data were analyzed with unconditional logistic regression and principal component analysis (PCA). RESULTS: Multivariate unconditional logistic regression analysis showed that the independent risk factors for nosocomial infection were use of the third generation cephalosporins for more than three days (odds ratio, OR 4.52, 95% confidence interval of OR 2.30 - 8.89), combined use of antibiotics (OR 2.86, 95% CI 1.51 - 5.43), use of quinolones for more than three days (OR 2.44, 95% CI 1.18 - 5.04), use of adrenal cortical hormone (OR 2.16, 95% CI 1.08 - 4.31) and oxygen inhalation (OR 2.56, 95% CI 1.14 - 5.72). Five principal components were extracted from the 14 risk factors for nosocomial infection with ESBLs-producing bacteria by principal component analysis, with a contribution of cumulative variance of 60.2%, and arranged in an order as follows, use of ventilator, tracheal intubation or tracheotomy, oxygen inhalation, retaining needle in vein, indwelling urethral catheter, use of the third generation cephalosporins over three days, hospitalization over ten days, use of quinolones over three days, combined use of antibiotics, use of aminoglycosides antibiotic over a week, use of adrenal cortical hormone, catheterized examination and prophylactic use of antibiotics. CONCLUSIONS: Nosocomial infection with ESBLs-producing bacteria could attribute to multiple factors, mainly to invasive manipulation and use of antibiotics. PMID- 12760796 TI - [Mutation of 5'noncoding region of the bcl-6 gene in diffuse large B cell lymphomas]. AB - OBJECTIVE: To observe the mutation of 5'noncoding region of bcl-6 gene in diffuse large B cell lymphoma (DLBCL) and its effect on lymphoma pathogenesis. METHODS: 38 DLBCL, 2 reactive hyperplasias, 5 follicular lymphomas and 5 T cell lymphomas were chosen for PCR direct sequence analysis using two sets of primers in 5'noncoding region of the bcl-6 gene. RESULTS: No mutation was found in the marginal region of reactive hyperplasias, T cell lymphomas, and follicular lymphomas but detected in 1/2 of the follicular center cells, and 7/38 cases of DLBCL. The incidence is less than that seen in other reports. Basepairs substitution and point insertion were the main mutation types. CONCLUSIONS: The positive rate of mutation of 5'noncoding region of bcl-6 gene in DLBCL is 18.7%, less frequent than the published data of DLBCL reported in other countries. It may, in some extent, participate in the pathogenesis and progression of DLBCL. PMID- 12760797 TI - [Study of B2 gene structure and its expression in colorectal cancer]. AB - OBJECTIVE: To get a complete cDNA sequence of B2 and evaluate the correlation on structure and expression between B2 and insulin like growth factor binding protein related protein 1 (IGFBP-rP1) in colorectal carcinomas, paired normal tissues, adenomas, tissues adjacent to the tumor, and colorectal carcinoma cell lines. METHODS: 5'RACE (rapid amplification of cDNA end) was applied to get the sequence of the 5' end of B2. Semi-quantitative RT-PCR and immunohistochemistry were used to detect the expression of B2 in colorectal cancer tissues and cell lines (SW480, SW1116, SW620, HCT8, CoLo205 and LoVo). RESULTS: A sequence of 1,125 bp was obtained by combining the sequence from 5'RACE product and the known sequence of B2. It shared 1,122/1,125 identities with IGFBP-rP1. At the level of mRNA, the expression of B2/IGFBP-rP1 was high in colorectal carcinomas, moderate in adenomas and tissues adjacent to tumor, low in normal tissues (P < 0.05). Five cell lines except SW480 showed no expression of B2/IGFBP-rP1. A significant difference was obtained in the immunoreactivity of B2/IGFBP-rP1 between normal tissue and cancer (P < 0.05). In 28.9% (22/76) samples, cancer cells locating at the invasive front of cancer nest had a stronger staining of B2/IGFBP-rP1 than those surrounding the lumen. These samples had also an increased frequency of lymph node metastases, increased depth of invasion and a stronger staining of B2/IGFBP-rP1 than in other samples (P < 0.05). CONCLUSIONS: B2 is the same gene as IGFBP-rP1. Overexpression of B2/IGFBP-rP1 may play an important role in the initiation and promotion of colorectal cancer. Its overexpression in invading tumor cells may be linking with an increased potential of invasion. PMID- 12760798 TI - [Loss of p53 gene and amplification of HER-2 oncogene in primary hepatocellular carcinoma and their clinical significance]. AB - OBJECTIVE: To investigate the deletion of p53 gene and amplification of HER-2 oncogene at chromosome 17 in primary hepatocellular carcinoma (HCC) and the clinical significance. METHODS: Interphase dual fluorescence in situ hybridization (FISH) was applied to detect the ratio of the number of p53 gene copy or HER-2 oncogene copy to that of chromosome 17 copy, to determine the p53 gene deletion and HER-2 oncogene amplification in nuclei prepared from 42 surgical specimens of HCC. Statistical analysis for their clinical significance was performed. RESULTS: Loss of p53 gene and amplification of HER-2 oncogene were detected in 27 (64.3%) and 9 (21.4%) of the 42 HCC respectively including 4 cases with low and 5 with high copy amplification. Six (14.3%) of 42 HCC showed simultaneously p53 gene deletion and HER-2 oncogene amplification. 61.9% (26/42) of HCC were polysomy 17, which correlated positively with p53 gene deletion (chi(2) = 12.286, P < 0.001). No close correlation between p53 gene loss and HER 2 oncogene amplification was found (chi(2) = 0.00, P = 1.00). Loss of p53 gene was related to the serum alpha-fetoprotein (AFP) level and the tumor size (P < 0.05). The postoperative 2-year survival rate (18.5%) of HCC patients with p53 gene deletion was significantly lower than postoperative 2-year survival rate (60.0%) of those without p53 gene loss (chi(2) = 7.467, P = 0.006). Meanwhile, HER-2 oncogene amplification showed a tendency of correlation with the tumor size (chi(2) = 2.973, P = 0.085), and the postoperative 2-year survival rate (0/9) of HCC patients with HER-2 oncogene amplification was significantly lower than those (42.4%) without HER-2 oncogene amplification (chi(2) = 3.977, P = 0.046). CONCLUSION: There were a high frequency of p53 gene deletion and a low frequency of HER-2 oncogene amplification in primary HCC, which might be involved in initiation and development of a subset of primary HCC. PMID- 12760799 TI - [Detection of gene promoter methylation and mRNA, protein expression levels of E cadherin in nasopharyngeal carcinoma]. AB - OBJECTIVE: To detect the gene promoter methylation, mRNA and protein expression levels of E-cadherin and beta-catenin in primary and metastatic tumor samples of nasopharyngeal carcinoma (NPC), and to investigate the mechanism of invasion and metastasis of neoplastic cell in NPC. METHODS: Twenty-one patients with NPC were studied. The samples of primary tumor and paired lymph node metastatic tumor were collected and examined for aberrant gene promoter methylation in E-cadherin by DNA Methylation-specific polymerase chain reaction (MSP). Reverse transcriptase polymerase chain reaction (RT-PCR), Western blotting and immunohistochemical staining were adopted to detect mRNA and protein levels of E-cadherin and beta catenin. RESULTS: (1) The gene promoter methylation in E-cadherin was 52.4% (11/21) in primary tumor of NPC, and 80.9% (17/21) in lymph node metastatic tumor, which existed significant difference (P < 0.05). (2) In primary tumor, about 80% (0 approximately 100%) neoplastic cells expressed E-cadherin protein on the average, which was significantly higher than that of metastatic tumor (50% on the average, P = 0.004). The expression levels of beta-catenin protein were high in both primary and metastatic tumors, but with no statistic difference (P = 0.698). (3) By Western blotting analysis, the relative intensity of protein expression in E-cadherin was significantly higher in primary tumor (206.7 +/- 32.7) compared to that of metastatic tumor (65.0 +/- 15.9), while the expression of beta-catenin protein showed no difference between them (P = 0.754). (4) mRNA expression level of E-cadherin was higher in primary tumor than that of metastatic tumor.No remarkable difference was found for the mRNA expression of beta-catenin. CONCLUSIONS: (1) Downregulation of mRNA and protein expression of E cadherin may play a critical role in neoplastic cell invasion and metastasis in NPC. The aberrant promoter methylation of E-cadherin may ultimately alter the mobility and scattering of tumor cells in NPC. (2) Downregulation of E-cadherin alone may be enough for the tumor cell to lose intercellular adhesions which results in tumor cell invasion and metastasis. However, mutant beta-catenin could also involve in this progress. (3) The detection of gene promoter hypermethylation of E-cadherin should be evaluated in the screening and surveillance of NPC. PMID- 12760800 TI - [Tissue transglutaminase protein expression in human brain tumors]. AB - OBJECTIVE: To investigate expression of tissue transglutaminase (tTG) protein and its role in carcinogenesis of brain tumors. METHODS: tTG protein was detected by immunohistochemical method in 62 astrocytomas, 18 oligodendrogliomas, 30 benign meningiomas, 30 pituitary adenomas and 10 normal brain tissues. RESULTS: (1) In brain tumors, tTG protein expression was heterogeneous locating in tumor and endothelial cells. (2) Immunoreactivity of tTG protein was significantly different between different grades of astrocytomas. (3) Expression intensity of tTG protein in glioma was higher than that in benign brain tumors. (4) Strong expression of tTG protein in tumor cell was obtained around the necrosis foci and apoptotic cells in astrocytomas. CONCLUSIONS: tTG protein expression contributed to tumor malignant progression in malignant brain tumors. PMID- 12760801 TI - [Relationship between mitochondrial DNA instability and interleukin-8 activity in gastric mucosa]. AB - OBJECTIVE: To evaluate the relationship between mitochondrial DNA instability (mtMSI) and interleukin-8 (IL-8) activity in gastric mucosa of various lesions. METHODS: IL-8 level in gastric mucosa was assayed using ELISA method. The mtMSI was detected by PCR-SSCP techniques. RESULTS: mtMSI was observed in 11 out of 30 (36.7%) gastric cancers, 2 of 15 (13.3%) intestinal metaplasia, 2 of 10 dysplasia and 1 of 10 chronic atrophic gastritis. IL-8 level in mtMSI+ group [(76.8 +/- 3.8) pg/mg] was significantly higher than that in mtMSI- group [(48.3 +/- 3.6) pg/mg, P < 0.05]. CONCLUSION: mtMSI closely correlates with IL-8 level in gastric mucosa and is involved in gastric carcinogenesis. PMID- 12760802 TI - [Primary gastric endocrine tumors]. AB - OBJECTIVE: To study the clinicopathological features of gastric neuroendocrine tumors. METHODS: Twenty cases were reviewed. The specimens were formalin-fixed, paraffin-embedded and immunostained by S-P method. RESULTS: Among the twenty cases, one case was carcinoid, three were malignant carcinoids, six had small cell carcinomas and ten had mixed extocrine--endocrine carcinomas. Immunohistological examination of tumor cells found 80% positive for S-100, NSE (85%), CgA (50%), SY (50%), gastrin (30%), serotonin (65%), AE1/AE3 (50%), and CEA (80%). CONCLUSIONS: In the WHO classification, there are five histological types in endocrine tumors of gastrointestinal tract. They are carcinoid, malignant carcinoid, small cell carcinoma, mixed exocrine--endocrine carcinoma and tumor-like lesions. But some cases in our paper were so different that they could not be classified. The gastric endocrine tumors are different from intestinal endocrine tumors and in classification, treatment and prognosis. PMID- 12760803 TI - [The role of hepatitis B virus X gene and p53 on hepatocellular carcinoma cell growth]. AB - OBJECTIVE: To explore the effects of hepatitis B virus X gene and p53 on hepatocellular growth. METHODS: Two kinds of plasmids containing sense and antisense human wild p53 gene respectively were constructed. SMMU-7721 cells were transfected with HBx, sense-wtp53 antisense-wtp53 separately or cotransfected with either HBx and sense-wtp53 or HBx and antisense-wtp53. Flow cytometry was adopted to measure the apoptosis rates and the effects of HBx on cell cycle progression. The activity of p21(Waf1) promoter-luciferase construct was detected. Growth curves for SMMU-7721 stably transfected with pcDNA3 and pcDNA3HBx were analyzed. RESULTS: After doxorubicin administration, HBx was noticed able to initiate apoptosis of the liver cells. The apoptosis rate was 5.32% in the pcDNA3 transfected and 12.66% in the pcDNA3HBx transfected groups respectively. HBx could also abrogate p53-mediated apoptosis. The apoptosis rate in groups transfected with pcDNA3, pcDNA3wtp53 and pcDNA3HBx + pcDNA3wtp53 was 5.32%, 11.72% and 4.67% respectively. In compared with the normal group, the number of cells in transiently HBx-expressed group and HBx-transfected group decreased 4.79% and 10.25% respectively. HBx inhibited the activity of p21(Waf1) promoter-luciferase constructed (P < 0.05) and promoted cell growth. The growth rate of HBx expression cells was faster. CONCLUSION: Under DNA damage, HBx reduced expression of p21(Waf1) by repressing the activity of p53 protein, followed by disturbing the regulation of G(0)-G(1) cell cycle checkpoint, and promoted the growth rate of hepatoma cells. PMID- 12760804 TI - [Effects of two variants of ING1 expression on tumor cell growth regulation]. AB - OBJECTIVE: To study effects of alternative transcripts of ING1 transfection on human cancer cell lines. METHODS: p47/ING1A and p33/ING1B expression vehicles were constructed and introduced into a human breast cancer cell line MCF-7 and a human lung cancer cell line PAa, both expressing wild-type p53 protein. Growth characteristics of the transfectants and potentially related genes were analyzed. RESULTS: The levels of p47/ING1A and p33/ING1B protein elevated respectively in tumor cells of MCF-7 and PAa after transfected with p47/ING1A and p33/ING1B, and the latter was much higher than that of the former. Ectopic overexpression of p33/ING1B effectively blocked tumor cell growth and arrested cells in the G(0) approximately G(1) phase of the cell cycle (P < 0.01), while p47/ING1A gave no effect on cell growth or cell cycle. Tumor cells overexpressing p33/ING1B contained more p21(WAF1) protein than that of the control cells, with undisturbed p53 protein level. CONCLUSIONS: Expression of two different transcripts of ING1 may have different effects on tumor cell growth. p33/ING1B may cooperate with p53 in stimulating expression of p21(WAF1) gene, thus to arrest cell cycle and to inhibit tumor cell growth. p33/ING1B may be considered to be a candidate as a partner of p53 in gene therapy. PMID- 12760806 TI - [Tear film function in non-insulin dependent diabetics]. AB - OBJECTIVE: To investigate whether diabetes mellitus is correlated with tear film dysfunction. METHODS: In 100 non-insulin dependent diabetic cases (200 eyes) and 80 non-diabetic controls, the tear break-up time was determined, and the staining with fluorescein sodium on the cornea and conjunctival impression cytology were performed. RESULTS: When compared with the healthy control group, the diabetic group showed decreased tear film break up time (u = 12.60, P < 0.01), increased rate of staining with fluorescein sodium on the cornea (chi(2) = 71.370, P < 0.01) and abnormal conjunctival epithelium. CONCLUSIONS: Non-insulin dependent diabetics are correlated with tear film dysfunction, so they are susceptible persons of the dry eye. The tear film break up time and staining with fluorescein sodium on the cornea should become routine tests of ophthalmology in diabetes mellitus. PMID- 12760805 TI - [Clinical characteristics of patients with dry eye syndrome]. AB - OBJECTIVE: To learn the clinical characteristics of patients with dry eye syndrome. METHODS: The following items were recorded in 115 patients (229 eyes) with dry eye, including symptoms, causation, systematic diseases, slit-lamp examination, tear break-up time, basal and reflex Schirmer's test, vital staining (fluorescent and rose bangle) and meibomian gland dysfunction examination. Rheumatoid factor and auto-antibody detection were performed in Sjogren's syndrome suspected patients. RESULTS: Aqueous tear deficiency (ATD, 48.7%) ranked the most common type, followed by over-evaporation dry eye (34.8%), mixed type (13.9%) and conjunctivochalasis (3.5%). In all the causes of the dry eye, about 11.3% had Sjogren syndrome (SS). Females suffering from dry eye were more than males, especially SS. Dryness was the most common symptom (84.0%), especially in ATD patients, then followed by ocular fatigue (72.0%), foreign body sensation (64.0%) and impairment of vision (56.0%). The ocular irritation was more severe in meibomain gland dysfunction (MGD) patients than in ATD patients. Among the results of tear break-up time (BUT), rose bangle (Rb) staining and fluorescent (Fl) staining in all types of dry eye, significant relationship was found among them, especially between Rb and Fl score (r = 0.612, P = 0.000). SS patients had much more severe abnormality in all the four signs than non-SS aqueous tear deficiency (NSTD) and MGD patients. However, in the comparisons of BUT, Rb and Fl between NSTD and MGD patients, there were no significant differences. CONCLUSION: Symptoms combined with examinations of BUT, Schirmer's test, Fl and Rb staining and meibomian gland function are the necessary means to diagnose most of the dry eye patients. PMID- 12760807 TI - [Preliminary clinical results of optical lamellar keratoplasty assisted by homemade-microkeratome]. AB - OBJECTIVE: To evaluate the efficacy and safety of optical lamellar keratoplasty assisted by homemade-microkeratome in treating corneal opacification for optical indications. METHODS: In 12 eyes of 11 patients with corneal opacities and healthy endothelium, the homemade-microkeratome was used for the resection of corneal opacities whose depth measured by optical pachymeter preoperatively, and the donor disc was excised by the same technique. After the resection of corneal opacities, the residual bed thickness was measured by ultrasonopachymeter. The disparities in thickness and diameter between the bed and the donor disc were reduced as least as possible by the use of different suction rings and microkeratome heads. The donor disc was secured with four or eight interrupted sutures. The period of follow-up ranged from 7 to 14 months. The transparency of cornea, donor-host interface and the refractive changes after surgery were examined by slit-lamp microscopy and Keratron videokeratoscope. RESULTS: The slit lamp microscopic examination showed no severe complications after surgery, all grafts appeared to be transparent without recognizable opacity in the interface except 1 eye with epithelial ingrowth and local graft melting subsequently. Visual acuities were improved in all eyes, 7 eyes had best corrected visual acuity better than 0.5 and 2 eyes reached 0.8. During the follow-up 3 to 6 months, the mean corneal refractive power significantly increased (2.36 +/- 1.25) D (P = 0.016), and the corneal astigmatism significantly decreased (0.97 +/- 0.95) D (P = 0.023) Compared with that before surgery, the increase of the corneal refractive power had negative correlation with the thickness of residual bed (r = -0.830, P = 0.004). CONCLUSIONS: This easy, safe new technique leads to good visual results and represents a good alternative to penetrating keratoplasty in treating corneal opacities. PMID- 12760808 TI - [The effect of TNF-related apoptosis-inducing ligand on in vivo graft survival after ex vivo adenovirus-mediated gene transfer to cornea]. AB - OBJECTIVE: To observe the effect of tumor necrosis factor-related apoptosis inducing ligand (TRAIL) on the allograft rejection after ex vivo adenovirus mediated gene transfer to cornea. METHODS: By co-culture with corneal graft, a replication-deficient adenovirus encoding the mTRAIL gene (Ad-TRAIL) was transduced into graft endothelium of mice. Immunohistochemistry was used to look for the TRAIL protein expression in ex vivo corneal endothelium. The grafts obtained from C57BL/6 mice that carried Ad-TRAIL were transplanted into the eyes of BALB/c mice. The occurrence time, the intensity of immunorejection and the immersion of CD4(+) and CD8(+) T lymphocytes were observed. TUNEL was used to detect the cell's apoptosis in the corneal grafts. We used uninfected group, Ad green-fluorescent protein (GFP) transduced group and soluble death receptor 5 (sDR5) group as controls. RESULTS: TRAIL protein expression was seen on more than 50% of endothelium 3 days after Ad-TRAIL transduced into corneal grafts in vitro, with higher expression for 2 weeks, then gradually reduced to negative till 3 weeks. The mean time of allograft rejection was 17.7 days in uninfected group, 22 days in Ad-TRAIL group, 17.4 days in Ad-GFP group and 12.3 days in sDR5 group. The differences among four groups were statistically significant. There were distributions of CD4(+), CD8(+) T lymphocytes and apoptotic cells in all rejected corneal grafts. CONCLUSIONS: TRAIL protein can inhibit corneal allograft rejection in mice and prolong the survival time of transplanted allografts. PMID- 12760809 TI - [Ultrastructural study of proprioceptors in extraocular muscles of congenital nystagmus subjects]. AB - OBJECTIVE: To observe the ultrastructure and the light microscopic structure alterations of the proprioceptors in the extraocular muscles (EOM) of congenital nystagmus (CN) subjects and discuss the probable effect and its mechanism of these alterations on CN to approach a new reasonable treatment for CN. METHODS: Twenty-one proprioceptors (muscle spindles) from 8 extraocular muscles of two typical CN were studied under both light microscope and transmission electron microscope. They were compared with 15 proprioceptors obtained from normal fresh cadavers. RESULTS: The proprioceptors in the EOM of CN subjects were smaller and their inner capsules became collapsed, discontinuous and vacuolated. At the same time, the muscle fibers and some endings of sensory nerves of EOMs from CN also had degenerative ultrastructural changes. CONCLUSIONS: The ultrastructure of the proprioceptors, muscle fibers and nerve endings in the EOMs of CN subjects are all changed compared with that of the controls. These alterations probably affect EOM's function and its proprioceptive input, which are probably related to the etiology and pathogenesis of CN. PMID- 12760810 TI - [A study of PDE6B gene mutation and phenotype in Chinese cases with retinitis pigmentosa]. AB - OBJECTIVE: To identify the mutation spectrum of phosphodiesterase beta subunit (PDE6B) gene, the incidence in Chinese patients with retinitis pigmentosa (RP) and their clinical phenotypic characteristics. METHODS: Screening of mutations within PDE6B gene was performed using polymerase chain reaction-heteroduplex single strand conformation polymorphism (PCR-SSCP) and DNA sequence in 35 autosomal recessive (AR) RP and 55 sporadic RP cases. The phenotypes of the patients with the gene mutation were examined and analyzed. RESULTS: Novel complex heterozygous variants of PDE6B gene in a sporadic case, a T to C transversion in codon 323 resulting in the substitution of Gly by Ser and 2 base pairs (bp: G and T) insert between the 27th-28th bp upstream of the 5'-end of exon 10 were both present in a same isolate RP. But they are not found in 100 unrelated healthy individuals. Ocular findings showed diffuse pigmentary retinal degeneration in the midperipheral and peripheral fundi, optic atrophy and vessel attenuation. Multi-focal ERG indicated that the rod function was more severely deteriorated. A mutation was found in a case with RP in a ARRP family, a G to A transversion at 19th base upstream 5'-end of exon 11 (within intron 10) of PDE6B gene. A sporadic RP carried a sequence variant of PDE6B gene, a G to C transition, at the 15th base adjacent to the 3'-end of exon l8. In another isolate case with RP was found 2 bp (GT) insert between 31st and 32nd base upstream 5'-end of exon 4 (in intron 3) of PDE6B gene. CONCLUSIONS: There are novel complex heterozygous mutations of PDE6B gene responsible for a sporadic RP patient in China. This gene mutation associated with rod deterioration and RP. Several DNA variants were found in introns of PDE6B gene in national population. PMID- 12760811 TI - [Application of capsular tension ring in cataract surgery for zonular dialysis]. AB - OBJECTIVE: To evaluate the application of capsular tension ring (CTR) in the cataract surgery for zonular dialysis. METHODS: Eleven cases of partially traumatized zonular dialysis patients were managed with CTR after continuous curvilinear capsulorrhexis (CCC), cataract extraction and posterior chamber intraocular lens (IOL) implantation. RESULTS: The IOLs in all the eyes were basically in the normal position without tilting or obvious decentration. One month postoperatively, the best-corrected visual acuity (BCVA) was 0.4 in 4 eyes, 0.5-0.8 in 5 eyes and over 0.8 in 2 eyes. A side-by-side overlap of both eyelets of the ring happened in 1 eye due to the bigger diameter of CTR, and no other complications were seen except the original complication of cataract surgery. CONCLUSIONS: For the patients of zonular dialysis, the CTR can preserve the integrity of the capsular bag, provide the necessary capsular space while giving a counter-traction, reduce asymmetric capsular forces, stabilize the vitreous base, facilitate phacoemulsification, aspiration of cortex and IOL implantation, prevent the postoperative IOL decentration and allow a quicker visual recovery. The CTR is a useful device that effectively supplements the currently available cataract surgery armamentarium, and develops the application of posterior chamber IOL implantation. PMID- 12760812 TI - [An analysis of measurement error in Orbscan topography system]. AB - OBJECTIVE: To discuss the measurement error and the influence factor of Orbscan Topography to provide more accurate data for clinical application. METHODS: Orbscan Topography System and A-scan ultrasonography were used to measure the corneal thickness and anterior chamber depths pre-operatively and post operatively, including 200 cases (400 eyes) undergoing laser in situ keratomileusis (Lasik) and 50 cases (100 eyes) undergoing photorefractive keratectomy (PRK). The curvature of posterior corneal surface was measured by Orbscan topography before and after the operations, and the result of measurement was compared. RESULTS: Pre-operatively, the value of corneal thickness was (18.83 - 24.29) microm lower measured by Orbscan topography than that by A ultrasonography, and post-operatively the value was (56.32 - 139.52) micro m lower than that by A-ultrasonography. The postoperative value was obviously higher than the pre-operative one. The value of central anterior chamber depth was (0.16 - 0.25) mm lower measured by Orbscan system than that by A ultrasonography pre-operatively and post-operatively. There was no clinical significance. The difference in the comparison of posterior corneal surface curvature before and after Lasik was -0.60 D measured by Orbscan System. The difference in the comparison of posterior corneal surface curvature before and after PRK was -0.51 D in the lower diopter group and -1.12 D in medium diopter group. CONCLUSIONS: It is necessary to improve the accuracy of Orbscan Topography System. The Haze and the change of anterior corneal surface curvature after Lasik and PRK are the main causes of measurement error in Orbscan Topography System. Clinically, it is an un-negligible important problem in the measurement of Orbscan system. PMID- 12760813 TI - [A study on cytokine levels and nitric oxide content in rabbit aqueous humor after lens extraction and intraocular lens implantation]. AB - OBJECTIVE: To investigate the relationship among inflammatory reaction and cytokine levels, nitric oxide (NO) content in aqueous humor after intraocular lens implantation. METHODS: Eighteen New Zealand rabbits were divided randomly into 3 groups (each 6 rabbits): (1) control group, (2) extracapsular cataract extraction group (ECCE) and (3) ECCE and posterior chamber intraocular lens implantation group (ECCE + IOL). The inflammation of all experimental rabbit eyes were observed by a zoom-photo slit-lamp microscope 0, 1, 3, 7, 14, 30 days postoperatively, including corneal edema and anterior chamber exudation. Meanwhile, aqueous humor was drawn for white blood cell (WBC) count and classification, as well as for NO(2)(-)/NO(3)(-) and cytokine assays, including interleukin-2 (IL-2), tumor necrosis factor-alpha (TNF-alpha). Statistics were taken by SPSS software. RESULTS: (1) The anterior chamber exudation was the most serious and monocyte/macrophage in aqueous humor were the highest in ECCE + IOL group in postoperative 7 - 14 days. (2) The levels of IL-2, TNF-alpha and the content of NO(2)(-)/NO(3)(-) in aqueous humor of ECCE + IOL group were higher than that in ECCE group and control group in the postoperative 1 - 14 days respectively, and they increased to their peak values at the postoperative 3 - 7 days and decreased gradually after postoperative two weeks. (3) The change regularity of IL-2, TNF-alpha, NO(2)(-)/NO(3)(-) and inflammatory reaction in each group were basically similar, i.e. the more serious the reaction, the higher the levels of the contents. CONCLUSION: The intraocular inflammation after intraocular lens implantation is closely related to the changes of cytokine levels and NO content in aqueous humor. PMID- 12760814 TI - Azithromycin versus Penicillin in Acute Group A Streptococcal Tonsillopharyngitis. PMID- 12760815 TI - Etiology and Management of Recurrent Aphthous Stomatitis. AB - Recurrent aphthous stomatitis (RAS) is an ulcerative condition that affects the oral mucosa without evidence of an underlying medical disorder. RAS is characterized by the appearance of round, shallow ulcerations surrounded by inflammation that chiefly involves the nonkeratinized mucosa. The etiology of RAS is unknown, but has a strong hereditary component and appears to be related to an immune reaction against the oral mucosa. RAS must be distinguished from other diseases that cause recurring oral ulcers such as Behcet's syndrome, systemic lupus erythematosus, and Crohn's disease. This paper reviews the current theories regarding the etiology of RAS, the clinical evaluation of patients with recurring aphthous ulcers, and describes current treatment options for this condition. PMID- 12760816 TI - The Utility of Radiologic Studies in the Diagnosis and Management of Rhinosinusitis. AB - The past two decades have seen the development of computed tomography scanning as the principal investigation for paranasal sinus disease. This article describes the benefits and limitations of computed tomography scanning in the assessment of rhinosinusitis. It also discusses the role of magnetic resonance imaging as a further diagnostic tool in the assessment of rhinosinusitis and in particular its complications. Other imaging modalities such as plain radiographs and ultrasound have limited use in this field. The introduction of image guidance techniques as an aid to surgical treatment and possible future developments are outlined. PMID- 12760818 TI - Progress in the Diagnosis, Prevention, and Treatment of Pertussis. AB - Pertussis ("whooping cough"), caused by the gram- negative pleomorphic bacillus Bordetella pertussis, is a highly contagious, potentially life-threatening respiratory tract illness that has re-emerged worldwide as a cause of substantial morbidity and mortality in infants, children, and adolescents, even in countries with high vaccination rates. Waning immunity after immunization during childhood has been associated with a growing pool of susceptible adolescents and adults who are capable of transmitting pertussis to vulnerable unvaccinated or incompletely vaccinated infants. The use of acellular pertussis vaccine boosters in adolescents has been proposed and is likely to be recommended. Active immunization and improved methods for early diagnosis are key in the management of pertussis, and represent the most rapidly evolving aspects of this disease. PMID- 12760817 TI - The Etiology, Pathophysiology, and Management of Otitis Media with Effusion. AB - Otitis media with effusion (OME) is a common and important condition that may result in developmental delay in children, and significant health care resources are devoted to its management. Newer techniques including polymerase chain reaction are implicating organisms not previously considered important in etiology. The role of gastroesophageal reflux as a cause of OME is likely to receive greater research attention. Regarding prevention, more is being learned about potentially modifiable risk factors such as environmental smoke, care outside the home, and breast feeding. Although immunization may to play a role in the future, existing evidence suggests that the general population of children should not be immunized in order to prevent OME. Several major studies have recently added to the understanding of epidemiology and management. Large trials in the United States, the Netherlands, and the UK suggest that OME is not an appropriate condition to include in a screening program. In addition, the advantages of early treatment with ventilation tubes over watchful waiting in terms of language development tend be modest and diminish by about 18 months. Treatment with hearing aids should be further evaluated. The search for effective medical management continues, and better ways are being identified of targeting interventions to those children with OME who are most likely to benefit. PMID- 12760819 TI - Microbiology and Treatment of Halitosis. AB - The many thousands of individuals who experience oral malodor that stems from the overgrowth of proteolytic, anaerobic bacteria on their tongue surfaces can be successfully treated by a regimen that includes tongue brushing and tooth brushing, often in combination with a mouthrinse containing an antibacterial agent. Several candidate mouthrinses containing essential oils (Listerine; Warner Lambert, Morris Plains, NJ), ZnCl(2), chlorine dioxide, or an oil:water cetylpyridium chloride mouthrinse have reduced the organoleptic scores of individuals with moderate levels of oral malodor in the absence of tongue brushing. Very little long-term data beyond 6 weeks of use are available. PMID- 12760820 TI - A Tuberculosis Treatment Completion Trial. PMID- 12760821 TI - Drug-resistant Streptococcus pneumoniae in Community-acquired Pneumonia. AB - The emergence of Streptococcus pneumoniae isolates resistant to not only penicillin, but to other antipneumococcal agents as well, has major public health implications. Drug-resistant S. pneumoniae are distributed worldwide, and resistance has become increasingly prevalent in the United States within the past decade. The relevance of resistance, particularly to the beta-lactams, to treatment outcome has been subject to debate. Pneumonia due to intermediate-level resistant penicillin-resistant isolates of S. pneumoniae appears to be adequately treated by beta-lactam agents. Interpretation of resistance reports, which may be based on achievable cerebrospinal fluid levels of drug, may depend on the clinical setting, and efforts are underway to adjust breakpoints so that reports are more easily applicable to clinical practice. Infectious Diseases Society of America and American Thoracic Society guidelines, as well as others, for community-acquired pneumonia have addressed the impact of drug-resistant S. pneumoniae on antimicrobial selection. PMID- 12760822 TI - Gram-negative Diplococcal Respiratory Infections. AB - Human respiratory tract infections caused by gram- negative diplococci continue to remain significant issues in health care. Although not addressed as frequently as the classical diplococcal pneumonia, the gram-positive Streptococcus pneumoniae (the pneumococcus), infections due to Neisseria meningitidis (the meningococcus), and Moraxella catarrhalis (formerly called both Neisseria catarrhalis and Branhamella catarrhalis) are addressed here including their microbiology, respiratory tract manifestations, antimicrobial treatment, and potential prevention with immunization. PMID- 12760823 TI - Urban Tuberculosis: The New Face of an Old Problem. AB - Tuberculosis (TB) persists as a worldwide health crisis. Nearly one third of the world's population is infected with TB and it remains a critical public health issue. In the past two decades, international efforts have focused on improving identification of those infected with TB as well as supporting molecular microbiologists in developing better diagnostic techniques. Furthermore, implementation of programs such as directly observed therapy have assisted more patients in receiving and completing therapy. This review aims to identify some of the more relevant findings in the field of TB over the past few years with a special emphasis on TB in urban communities. PMID- 12760824 TI - Cytomegalovirus Disease in the Highly Active Antiretroviral Therapy Era. AB - Cytomegalovirus (CMV) is a major cause of morbidity and mortality in AIDS patients. Epidemiologic studies indicate that until 10 years ago, nearly one half of HIV-infected patients eventually developed CMV end-organ disease, including chorioretinitis, esophagitis, colitis, pneumonia, and central nervous system disease. Since the introduction of highly active antiretroviral therapy (HAART) this incidence has declined dramatically. Nonetheless, patients still present with CMV disease and resistance or intolerance to HAART does develop, which may give rise to a resurgence of CMV syndromes in AIDS patients. Until recently, only intravenous ganciclovir and foscarnet were available for management of CMV infection. With the advent of additional agents, clinicians now face the challenge of optimizing therapy for individual patients. This paper reviews the most common clinical syndromes caused by CMV, the treatment options, as well as an approach to diagnosing and treating antiviral resistance. PMID- 12760825 TI - Immune Restoration Inflammatory Syndromes: The Dark Side of Successful Antiretroviral Treatment. AB - The prevalence of cellular immunodeficiency has increased due to rising use of immunosuppressive therapies and the pandemic spread of HIV infection. More recently, the introduction of highly active antiretroviral therapy (HAART) in HIV has led to significant immune reconstitution, even in patients with previously long-lasting secondary immunodeficiency. HAART reduces morbidity and mortality in HIV infection and also changes the clinical course of prevalent subclinical opportunistic infections or autoimmune diseases. Atypical inflammatory disorders develop after initiation of HAART and have been summarized as "immune reconstitution syndrome," "immune restoration disease," and "immune restoration inflammatory syndrome." However, diagnostic criteria and standards of therapy are yet to be defined. The awareness for these diseases is of increasing importance from a clinical point of view. This review summarizes the variety of immunoreconstitution disorders and describes possible diagnostic pitfalls. We also propose possible therapeutic options. PMID- 12760826 TI - The Comparative Toxicogenomics Database (CTD). AB - The Mount Desert Island Biological Laboratory in Salsbury Cove, Maine, USA, is developing the Comparative Toxicogenomics Database (CTD), a community-supported genomic resource devoted to genes and proteins of human toxicologic significance. CTD will be the first publicly available database to a) provide annotated associations among genes, proteins, references, and toxic agents, with a focus on annotating data from aquatic and mammalian organisms; b) include nucleotide and protein sequences from diverse species; c) offer a range of analysis tools for customized comparative studies; and d) provide information to investigators on available molecular reagents. This combination of features will facilitate cross species comparisons of toxicologically significant genes and proteins. These comparisons will promote understanding of molecular evolution, the significance of conserved sequences, the genetic basis of variable sensitivity to environmental agents, and the complex interactions between the environment and human health. CTD is currently under development, and the planned scope and functions of the database are described herein. The intent of this report is to invite community participation in the development of CTD to ensure that it will be a valuable resource for environmental health, molecular biology, and toxicology research. PMID- 12760836 TI - Model selection in genomics. PMID- 12760830 TI - Genomic and proteomic profiling of responses to toxic metals in human lung cells. AB - Examining global effects of toxic metals on gene expression can be useful for elucidating patterns of biological response, discovering underlying mechanisms of toxicity, and identifying candidate metal-specific genetic markers of exposure and response. Using a 1,200 gene nylon array, we examined changes in gene expression following low-dose, acute exposures of cadmium, chromium, arsenic, nickel, or mitomycin C (MMC) in BEAS-2B human bronchial epithelial cells. Total RNA was isolated from cells exposed to 3 M Cd(II) (as cadmium chloride), 10 M Cr(VI) (as sodium dichromate), 3 g/cm2 Ni(II) (as nickel subsulfide), 5 M or 50 M As(III) (as sodium arsenite), or 1 M MMC for 4 hr. Expression changes were verified at the protein level for several genes. Only a small subset of genes was differentially expressed in response to each agent: Cd, Cr, Ni, As (5 M), As (50 M), and MMC each differentially altered the expression of 25, 44, 31, 110, 65, and 16 individual genes, respectively. Few genes were commonly expressed among the various treatments. Only one gene was altered in response to all four metals (hsp90), and no gene overlapped among all five treatments. We also compared low dose (5 M, noncytotoxic) and high-dose (50 M, cytotoxic) arsenic treatments, which surprisingly, affected expression of almost completely nonoverlapping subsets of genes, suggesting a threshold switch from a survival-based biological response at low doses to a death response at high doses. PMID- 12760837 TI - Toxicogenomics: roadblocks and new directions. PMID- 12760838 TI - Phenotypic anchoring: linking cause and effect. PMID- 12760839 TI - Data explosion: bringing order to chaos with bioinformatics. PMID- 12760840 TI - How to review a paper. AB - Most scientists acquire their training in manuscript review not through instruction but by actually doing it. Formal training in manuscript analysis is rarely, if ever, provided. Editors usually choose reviewers because of expertise in a given subject area and availability. If an individual repeatedly submits bad reviews, it is likely that that person will not be asked to review a manuscript again. Being invited to review a manuscript is an honor, not only because you are being recognized for your eminence in a particular area of research but also because of the responsibility and service you provide to the journal and scientific community. The purpose of this article is to define how best to peer review an article. We will stipulate several principles of peer review and discuss some of the main elements of a good manuscript review, the basic responsibilities of a reviewer, and the rewards and responsibilities that accompany this process. Proper reviewer conduct is essential for making the peer review process valuable and the journal trustworthy. PMID- 12760841 TI - Hypertension module: an interactive learning tool in physiology. AB - The aim of the present study was to evaluate the strong or weak aspects of an interactive study module introduced during the "Cardiovascular and Respiratory Systems Subject Committee" in the second year of the medical program. Five study groups consisting of 25 students attended two-hour module sessions for six weeks with the same tutor. According to the module assessment questionnaire, the majority of the students assessed the module as excellent or good. The students reported that they had gained not only in knowledge but also in skills development. The general opinion of the students was that both the organization and the implementation of the module met their expectations. Nearly one-half of the students reported that their expectations with regard to the educational environment and the participation of students were fully met. The major weakness in this new educational trial appears to be assessment of the module. PMID- 12760842 TI - How do learning issues relate with content in a problem-based learning pathophysiology course? AB - The relation between learning process and content coverage is becoming increasingly important for the understanding of the effects of problem-based learning (PBL) on students' learning. In our medical school, PBL is used as a major educational strategy in the discipline of pathophysiology. A computer program was developed allowing students to register learning issues identified as needed during tutorial sessions and learning issues stated as covered during the individual study periods. In our study, we compared "planned" (learning issues identified during PBL sessions) and "accomplished" learning issues (covered after the independent study periods) identified by pathophysiology students from three consecutive years. We found that the planned learning issues raised during tutorial sessions related to the issues effectively accomplished during the independent study and that their number grew stepwise from basic to preclinical to clinical sciences. Pathophysiology was, globally, the most mentioned discipline. Moreover, the most mentioned disciplines from the basic, preclinical, and clinical areas were physiology, histopathology, and internal medicine, respectively. The single-discipline approach did not limit the student's capacity to identify and cover learning issues beyond the objectives of pathophysiology. PMID- 12760843 TI - Measurement of serum leptin concentrations in university undergraduates by competitive ELISA reveals correlations with body mass index and sex. AB - Synthesized mainly in adipocytes, leptin is a peptide hormone that plays a key role in the regulation of body weight and composition. The serum leptin concentrations of 193 Singapore university medical and bioscience undergraduates aged 19-26 yr were measured using a competitive ELISA kit, and their leptin levels were correlated with sex and body mass index (BMI). Mean leptin levels were more than twice as high in females than in males of corresponding weight status, especially among females of healthy weight who exhibited levels that were 5.7 times higher. Overweight individuals generally demonstrated higher circulating leptin concentrations than healthy-weight and underweight participants. The differences in mean leptin levels between underweight and overweight males (P = 0.006), as well as between healthy-weight and overweight males (P = 0.011) were statistically significant. Comparison tests of leptin levels between healthy-weight and underweight females were highly significant (P = 0.001). Highly significant linear correlations between BMI and the logarithm of leptin concentration were observed in the female (r = 0.44) and male (r = 0.36) groups. These results reiterate the impact of gonadal steroids as mediators of the apparent sexual dimorphism in circulating leptin. The findings also corroborate evidence that adiposity determines leptin levels. This laboratory exercise has educational value for undergraduates by determining their BMIs, by alluding to the importance of maintaining healthy body composition, and by emphasizing the molecular mechanisms of body weight regulation and obesity, with special reference to leptin. This practical study also exemplifies the principles and applications of the competitive ELISA technique and integrates certain key concepts of physiology, molecular biology, immunology, and medicine. PMID- 12760844 TI - Understanding the physiology of glutamate receptors by use of a protocol for neuronal staining. AB - Teaching students about the physiology of neurotransmitter receptors usually requires practical lessons with the use of sophisticated equipment and complex analysis of data. Here, we report our experience in teaching medical students with a simple, practical protocol that transforms the physiology of glutamate receptors into neuronal staining, observable under bright-field microscopy. Essentially, the students were challenged to selectively stain a subpopulation of cultured neurons expressing Ca(2+)-permeable alpha-amino-3-hydroxy-5 methylisoxazole-4-propionate (AMPA) receptors (a subgroup of ionotropic glutamate receptors). Neurons expressing this type of receptors were loaded with Co(2+) (in substitution for Ca(2+)) after nondesensitizing activation of AMPA receptors. After precipitation, the Co(2+) was revealed after treatment with silver. At the end of the procedure, the neurons expressing Ca(2+)-permeable AMPA receptors were visually identified under bright-field microscopy. The procedure allowed the visualization of the complete dendritic network of the stained neurons and allowed the students to learn very efficiently about the physiology of glutamate receptors. PMID- 12760845 TI - Pulmonary ventilation teaching aid: part 2. PMID- 12760846 TI - A model for visualizing fluid handling by the gastrointestinal tract. PMID- 12760849 TI - Variable sensitivity to bacterial methionyl-tRNA synthetase inhibitors reveals subpopulations of Streptococcus pneumoniae with two distinct methionyl-tRNA synthetase genes. AB - As reported previously (J. R. Jarvest et al., J. Med. Chem. 45:1952-1962, 2002), potent inhibitors (at nanomolar concentrations) of Staphylococcus aureus methionyl-tRNA synthetase (MetS; encoded by metS1) have been derived from a high throughput screening assay hit. Optimized compounds showed excellent activities against staphylococcal and enterococcal pathogens. We report on the bimodal susceptibilities of S. pneumoniae strains, a significant fraction of which was found to be resistant (MIC, > or =8 mg/liter) to these inhibitors. Using molecular genetic techniques, we have found that the mechanism of resistance is the presence of a second, distantly related MetS enzyme, MetS2, encoded by metS2. We present evidence that the metS2 gene is necessary and sufficient for resistance to MetS inhibitors. PCR analysis for the presence of metS2 among a large sample (n = 315) of S. pneumoniae isolates revealed that it is widespread geographically and chronologically, occurring at a frequency of about 46%. All isolates tested also contained the metS1 gene. Searches of public sequence databases revealed that S. pneumoniae MetS2 was most similar to MetS in Bacillus anthracis, followed by MetS in various non-gram-positive bacterial, archaeal, and eukaryotic species, with streptococcal MetS being considerably less similar. We propose that the presence of metS2 in specific strains of S. pneumoniae is the result of horizontal gene transfer which has been driven by selection for resistance to some unknown class of naturally occurring antibiotics with similarities to recently reported synthetic MetS inhibitors. PMID- 12760848 TI - Macrolide resistance by ribosomal mutation in clinical isolates of Streptococcus pneumoniae from the PROTEKT 1999-2000 study. AB - Sixteen (1.5%) of the 1,043 clinical macrolide-resistant Streptococcus pneumoniae isolates collected and analyzed in the 1999-2000 PROTEKT (Prospective Resistant Organism Tracking and Epidemiology for the Ketolide Telithromycin) study have resistance mechanisms other than rRNA methylation or efflux. We have determined the macrolide resistance mechanisms in all 16 isolates by sequencing the L4 and L22 riboprotein genes, plus relevant segments of the four genes for 23S rRNA, and the expression of mutant rRNAs was analyzed by primer extension. Isolates from Canada (n = 4), Japan (n = 3), and Australia (n = 1) were found to have an A2059G mutation in all four 23S rRNA alleles. The Japanese isolates additionally had a G95D mutation in riboprotein L22; all of these originated from the same collection center and were clonal. Three of the Canadian isolates were also clonal; the rest were not genetically related. Four German isolates had A2059G in one, two, and three 23S rRNA alleles and A2058G in two 23S rRNA alleles, respectively. An isolate from the United States had C2611G in three 23S rRNA alleles, one isolate from Poland had A2058G in three 23S rRNA alleles, one isolate from Turkey had A2058G in four 23S rRNA alleles, and one isolate from Canada had A2059G in two 23S rRNA alleles. Erythromycin and clindamycin resistance gradually increased with the number of A2059G alleles, whereas going from one to two mutant alleles caused sharp rises in the azithromycin, roxithromycin, and rokitamycin MICs. Comparisons of mutation dosage with rRNA expression indicates that not all alleles are equally expressed. Despite their high levels of macrolide resistance, all 16 isolates remained susceptible to the ketolide telithromycin (MICs, 0.015 to 0.25 microg/ml). PMID- 12760850 TI - Susceptibilities to levofloxacin in Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis clinical isolates from children: results from 2000-2001 and 2001-2002 TRUST studies in the United States. AB - Among respiratory tract isolates of Streptococcus pneumoniae from children, resistance to penicillins, cephalosporins, macrolides, and trimethoprim sulfamethoxazole (SXT) increases on an annual basis. Pediatric patients who do not respond to conventional therapy for respiratory tract infections someday may be treated with fluoroquinolones. In this study, MICs of beta-lactams, azithromycin, SXT, and levofloxacin were determined and interpreted by using NCCLS guidelines for isolates of S. pneumoniae (2,834 from children and 10,966 from adults), Haemophilus influenzae (629 from children and 2,281 from adults), and Moraxella catarrhalis (389 from children and 1,357 from adults) collected during the 2000-2001 and 2001-2002 respiratory illness seasons in the United States as part of the ongoing TRUST surveillance studies. Rates of resistance to penicillin, azithromycin, and SXT were > or = 7.5% higher among patients < or = 4 years old than among patients 5 to 10, 11 to 17, and > or = 18 years old in both the 2000-2001 and the 2001-2002 respiratory illness seasons. Levofloxacin resistance was detected in 2 of 2,834 isolates (0.07%) from patients <18 years old. Levofloxacin MICs of 0.25 to 1 micro g/ml accounted for 99.6, 99.5, 99.3, 99.7, 98.4, and 98.0% of isolates from patients < 2, 2 to 4, 5 to 10, 11 to 17, 18 to 64, and > 64 years old. Multidrug resistance was twice as common among patients < or = 4 years old (25.3%) as among patients 5 to 10 years old (13.7%), 11 to 17 years old (11.9%), 18 to 64 years old (12.1%), and > 64 years old (12.4%). The most common multidrug resistance phenotype in S. pneumoniae isolates for all age groups was resistance to penicillin, azithromycin, and SXT (70.3 to 76.6%). For H. influenzae and M. catarrhalis isolates from patients < 2, 2 to 4, 5 to 10, 11 to 17, 18 to 64, and > 64 years old, levofloxacin MICs at which 90% of the isolates were inhibited were 0.015 and 0.03 to 0.06 microg/ml, respectively, in the 2000-2001 and 2001-2002 respiratory illness seasons. In the 2000-2001 and 2001-2002 respiratory illness season surveillance studies in the United States, 99.9% of pediatric isolates of S. pneumoniae were susceptible to levofloxacin. If fluoroquinolones become a treatment option for pediatric patients, careful monitoring of fluoroquinolone susceptibilities will be increasingly important in future surveillance studies. PMID- 12760851 TI - Oral bioavailability and in vivo efficacy of the helicase-primase inhibitor BILS 45 BS against acyclovir-resistant herpes simplex virus type 1. AB - This study investigated the oral bioavailability and efficacy of BILS 45 BS, a selective herpes simplex virus (HSV) helicase-primase inhibitor, against acyclovir (ACV)-resistant (ACV(r)) infections mediated by the HSV type 1 (HSV-1) dlsptk and PAA(r)5 mutant strains. In vitro, the compound was more potent than ACV against wild-type clinical and laboratory HSV-1 strains and ACV(r) HSV isolates, as determined by a standard plaque reduction assay, with a mean 50% effective concentration of about 0.15 microM. The oral bioavailability of BILS 45 BS in hairless mice was 49%, with a peak concentration in plasma of 31.5 microM after administration of a single dose of 25 mg/kg. Following cutaneous infection of nude mice, both the HSV-1 dlsptk and PAA(r)5 mutant strains induced significant, reproducible, and persistent cutaneous lesions that lasted for more than 2 weeks. Oral treatment with ACV (100 or 125 mg/kg/day, three times a day by gavage) did not affect either mutant-induced infection. In contrast, BILS 45 BS at an oral dose of 100 mg/kg/day almost completely abolished cutaneous lesions mediated by both ACV(r) HSV-1 mutants. The 50% effective doses of BILS 45 BS were 56.7 and 61 mg/kg/day against dlsptk- and PAA(r)5-induced infections, respectively. Taken together, our results demonstrate very effective oral therapy of experimental ACV(r) HSV-1 infections in nude mice and support the potential use of HSV helicase-primase inhibitors for the treatment of nucleoside-resistant HSV disease in humans. PMID- 12760853 TI - Voriconazole inhibits fungal growth without impairing antigen presentation or T cell activation. AB - Invasive aspergillosis (IA) is the most common life-threatening invasive mold infection worldwide. The principal therapy for IA is amphotericin B, despite its known toxicity and immunosuppressive side effects. Studies in animal models of IA suggest a role for T lymphocytes in the pathology of the disease, although the precise role for Aspergillus-specific T cells remains undefined. The isolation and characterization of T lymphocytes in animal models of IA are hampered by the rapid outgrowth of the fungus in cultures derived from infected organs. In the present study, we tested the abilities of the antifungal drugs caspofungin acetate and voriconazole to inhibit fungal growth in vitro as a means of maintaining cultures of T cells from Aspergillus-infected mice. We demonstrate that while both antifungal drugs are inhibitory, only voriconazole completely inhibited fungal growth, allowing long-term maintenance of T-cell cultures. In addition, voriconazole had no inhibitory effect on the activation and maturation of dendritic cells or the proliferation of T lymphocytes. Thus, voriconazole appears to be a promising agent for use in in vitro studies of Aspergillus specific T lymphocytes in animal models of IA. PMID- 12760854 TI - Worldwide assessment of linezolid's clinical safety and tolerability: comparator controlled phase III studies. AB - Linezolid, an oxazolidinone antibiotic, has 100% oral bioavailability and favorable activities against gram-positive pathogens including multidrug resistant staphylococci, enterococci, and pneumococci. Safety assessments were conducted for 2,046 linezolid-treated patients and 2,001 comparator drug-treated patients from seven controlled clinical trials comparing the activities of linezolid and comparator drugs against nosocomial and community-acquired pneumonia, skin and skin structure infections, and methicillin-resistant staphylococcal infections. Drug-related adverse events were primarily transient. The most frequent (> or = 2%) adverse events caused by linezolid and the comparator drugs were diarrhea (4.3 and 3.2%, respectively; P = 0.074), nausea (3.4 and 2.3%, respectively; P = 0.036), and headache (2.2 and 1.3%, respectively; P = 0.047). Treatment discontinuations due to drug-related events (2.4 and 1.9%, respectively), serious adverse events (11.4 and 10.6%, respectively), and deaths (4.8 and 4.9%, respectively) were similar. No clinically significant drug-related hematologic events were reported, and laboratory safety data were comparable. In the first 6 months of postmarketing surveillance, hematologic abnormalities were reported in 0.1% of linezolid treated patients, but no irreversible blood dyscrasias were documented. The risk for transient, reversible hematologic effects from treatment with linezolid should be considered together with the clinical benefits associated with its use. PMID- 12760852 TI - Ciclopirox olamine treatment affects the expression pattern of Candida albicans genes encoding virulence factors, iron metabolism proteins, and drug resistance factors. AB - The hydroxypyridone ciclopirox olamine belongs to the antimycotic drugs used for the treatment of superficial mycoses. In contrast to the azoles and other antimycotic drugs, its specific mode of action is only poorly understood. To investigate the mode of action of ciclopirox olamine on fungal viability, pathogenicity, and drug resistance, we examined the expression patterns of 47 Candida albicans genes in cells grown in the presence of a subinhibitory concentration (0.6 micro g/ml) of ciclopirox olamine by reverse transcription PCR. In addition, we used suppression-subtractive hybridization to further identify genes that are up-regulated in the presence of ciclopirox olamine. The expression of essential genes such as ACT1 was not significantly modified in cells exposed to ciclopirox olamine. Most putative and known virulence genes such as genes encoding secreted proteinases or lipases had no or only moderately reduced expression levels. In contrast, exposure of cells to ciclopirox olamine led to a distinct up- or down-regulation of genes encoding iron permeases or transporters (FTR1, FTR2, FTH1), a copper permease (CCC2), an iron reductase (CFL1), and a siderophore transporter (SIT1); these effects resembled those found under iron-limited conditions. Addition of FeCl(3) to ciclopirox olamine-treated cells reversed the effect of the drug. Addition of the iron chelator bipyridine to the growth medium induced similar patterns of expression of distinct iron regulated genes (FTR1, FTR2). While serum-induced yeast-to-hyphal phase transition of C. albicans was not affected in ciclopirox olamine-treated cells in the presence of subinhibitory conditions, a dramatic increase in sensitivity to oxidative stress was noted, which may indicate the reduced activities of iron containing gene products responsible for detoxification. Although the Candida drug resistance genes CDR1 and CDR2 were up-regulated, no change in resistance or increased tolerance could be observed even after an incubation period of 6 months. This was in contrast to control experiments with fluconazole, in which the MICs for cells incubated with this drug had noticeably increased after 2 months. These data support the view that the antifungal activity of ciclopirox olamine may at least be partially caused by iron limitation. Furthermore, neither the expression of certain multiple-drug resistance genes nor other resistance mechanisms caused C. albicans resistance to this drug even after long-term exposure. PMID- 12760855 TI - Antimicrobial resistance of invasive pneumococci in Finland in 1999-2000. AB - The resistance patterns and macrolide resistance mechanisms of 910 Finnish invasive pneumococci isolated during 1999 and 2000 were studied. Macrolide resistance was detected in 6.9% of isolates. Penicillin resistance was detected in 1.5% of isolates, and penicillin intermediate resistance was detected in 4.0% of isolates. Active macrolide efflux, mediated by the mef(A) gene, was the most common macrolide resistance mechanism. Four macrolide-resistant isolates had mutations in rRNA or ribosomal protein L22. PMID- 12760856 TI - Tanshinone (Salviae miltiorrhizae extract) preparations attenuate aminoglycoside induced free radical formation in vitro and ototoxicity in vivo. AB - Antioxidant therapy protects against aminoglycoside-induced ototoxicity in animal models. A clinically suitable antioxidant must not affect the therapeutic efficacy of aminoglycosides or exhibit any side effects of its own. In addition, the treatment should be inexpensive and convenient in order to be implemented in developing countries where the use of aminoglycosides is most common. Standardized Salviae miltiorrhizae extracts (Danshen) are used clinically in China and contain diterpene quinones and phenolic acids with antioxidant properties. We combined in vitro and in vivo approaches to investigate the effect of a clinically approved injectable Danshen solution on aminoglycoside-induced free radical generation and ototoxicity. In vitro, Danshen inhibited gentamicin dependent lipid peroxidation (formation of conjugated dienes from arachidonic acid), as well as the gentamicin-catalyzed formation of superoxide (in a lucigenin-based chemiluminescence assay) and hydroxyl radicals (oxidation of N,N dimethyl-p-nitrosoaniline). Danshen extracts were then administered to adult CBA mice receiving concurrent treatment with kanamycin (700 mg/kg of body weight twice daily for 15 days). Auditory threshold shifts induced by kanamycin (approximately 50 dB) were significantly attenuated. Danshen did not reduce the levels in serum or antibacterial efficacy of kanamycin. These results suggest that herbal medications may be a significantly underexplored source of antidotes for aminoglycoside ototoxicity. Such traditional medicines are widely used in many developing countries and could become an easily accepted and inexpensive protective therapy. PMID- 12760857 TI - Effects of pyrimidine and purine analog combinations in the duck hepatitis B virus infection model. AB - To design new strategies of antiviral therapy for chronic hepatitis B, we have evaluated the antiviral activity of the combination of amdoxovir (DAPD), emtricitabine [(-)FTC], and clevudine (L-FMAU) in the duck hepatitis B virus (DHBV) model. Using their triphosphate (TP) derivatives in a cell-free system expressing a wild-type active DHBV reverse transcriptase (RT), the three dual combinations exhibited a greater additive inhibitory effect on viral minus-strand DNA synthesis than the single drugs, according to the Bliss independence model. Both dual combinations with DAPD TP were the most efficient while the triple combination increased the inhibitory effect on the DHBV RT activity in comparison with the dual association, however, without additive effect. Postinoculation treatment of experimentally infected primary duck hepatocytes showed that dual and triple combinations potently inhibited viral DNA synthesis during treatment but did not inhibit the reinitiation of viral DNA synthesis after treatment cessation. Preinoculation treatment with the same combinations exhibited antiviral effects on intracellular viral DNA replication, but it was unable to prevent the initial covalently closed circular DNA (cccDNA) formation. Short-term in vivo treatment in acutely infected ducklings showed that the dual combinations were more-potent inhibitors of virus production than the single treatments, with the L-FMAU and FTC combination being the most potent. A longer administration of L-FMAU and FTC for 4 weeks efficiently suppressed viremia and viral replication. However, no viral clearance from the liver was observed, suggesting that the enhanced antiviral effect of this combination was not sufficient for cccDNA suppression and HBV eradication from infected cells. PMID- 12760858 TI - Pharmacokinetics and pharmacodynamics of cefepime in patients with various degrees of renal function. AB - This study evaluated the pharmacokinetics of cefepime in 36 patients with different levels of renal function. Pharmacokinetic and pharmacodynamic parameters were calculated using samples obtained at steady state. Patients with creatinine clearance (CL(CR)) of >100 ml/min had more rapid clearance (CL) and a lower minimum concentration in serum (C(min)). C(min) in this group was found to be 3.3 +/- 3.6 mg/liter (mean and standard deviation), compared to 19.5 +/- 21.5 mg/liter in patients with a CL(CR) of between 60 and 100 ml/min (P = 0.025) and 14.0 +/- 11.5 mg/liter in patients with a CL(CR) of <60 ml/min (P = 0.009). Patient data were also analyzed by the nonparametric expectation maximization method and Bayesian forecasting. The median volume of distribution in the central compartment was 27.08 liters. CL and CL(CR) were highly correlated (P = 0.00033) according to the equation CL= 0.324 liters/h + (0.0551 x CL(CR)). The median rate constants from the central compartment to the peripheral compartment and from the peripheral compartment to the central compartment were 12.58 and 41.09 h(-1), respectively. The time-concentration profiles for 1,000 patients (CL(CR)s, 120, 60, and 30 ml/min) each receiving various dosing regimens were simulated by using Monte Carlo simulations. Standard dosing resulted in a C(min) that was greater than or equal to the MIC in more than 80% of the simulated profiles with MICs < or = 2 mg/liter. Current dosing recommendations may be suboptimal for monotherapy of infections due to less susceptible pathogens (e.g., those for which MICs are > or = 4 mg/liter), particularly when CL(CR) exceeds 120 ml/min. PMID- 12760859 TI - Clinical pharmacokinetics of cefamandole and ceftazidime administered by continuous intravenous infusion. AB - In view of the results of animal studies as well as theoretical considerations, continuous administration of beta-lactam antibiotics should be superior to intermittent administration because of the close relationship between efficacy and the duration of time in which the concentration of unbound antibiotics in plasma remains above the MIC. The aim of the present study was to establish the pharmacokinetic parameters of cefamandole and ceftazidime for patients receiving these cephalosporins by continuous infusion. The interindividual differences in the concentrations in plasma at the steady state were mainly attributable to variations in renal function, as estimated by the rate of creatinine clearance. Using these results, we derived formulas for both cephalosporins that can be used to determine on an individual basis the total daily dose needed to obtain a therapeutic concentration in plasma. These formulas were tested with a group of subsequent patients and proved to be practical and fairly reliable. For some patients, a correction for a possible underestimation of the renal clearance at presentation might be required. PMID- 12760860 TI - Antimicrobial resistance in respiratory tract Streptococcus pneumoniae isolates: results of the Canadian Respiratory Organism Susceptibility Study, 1997 to 2002. AB - A total of 6,991 unique patient isolates of Streptococcus pneumoniae were collected from October 1997 to June 2002 from 25 medical centers in 9 of the 10 Canadian provinces. Among these isolates, 20.2% were penicillin nonsusceptible, with 14.6% being penicillin intermediate (MIC, 0.12 to 1 microg/ml) and 5.6% being penicillin resistant (MIC, > or =2 microg/ml). The proportion of high-level penicillin-resistant S. pneumoniae isolates increased from 2.4 to 13.8% over the last 3 years of the study, and the proportion of multidrug-resistant S. pneumoniae isolates increased from 2.7 to 8.8% over the 5-year period. Resistant rates (intermediate and resistant) among non-beta-lactam agents were as follows: macrolides, 9.6 to 9.9%; clindamycin, 3.8%; doxycycline, 5.5%; chloramphenicol, 3.9%; and trimethoprim-sulfamethoxazole, 19.0%. Rates of resistance to non-beta lactam agents were higher among penicillin-resistant strains than among penicillin-susceptible strains. No resistance to vancomycin or linezolid was observed; however, 0.1% intermediate resistance to quinupristin-dalfopristin was observed. The rate of macrolide resistance (intermediate and resistant) increased from 7.9 to 11.1% over the 5 years. For the fluoroquinolones, the order of activity based on the MICs at which 50% of isolates are inhibited (MIC(50)s) and the MIC(90)s was gemifloxacin > clinafloxacin > trovafloxacin > moxifloxacin > grepafloxacin > gatifloxacin > levofloxacin > ciprofloxacin. The investigational compounds ABT-773 (MIC(90), 0.008 microg/ml), ABT-492 (MIC(90), 0.015 microg/ml), GAR-936 (tigecycline; MIC(90), 0.06 microg/ml), and BMS284756 (garenoxacin; MIC(90), 0.06 micro g/ml) displayed excellent activities. Despite decreases in the rates of antibiotic consumption in Canada over the 5-year period, the rates of both high-level penicillin-resistant and multidrug-resistant S. pneumoniae isolates are increasing in Canada. PMID- 12760861 TI - Antimicrobial resistance in Haemophilus influenzae and Moraxella catarrhalis respiratory tract isolates: results of the Canadian Respiratory Organism Susceptibility Study, 1997 to 2002. AB - A total of 7,566 unique patient isolates of Haemophilus influenzae and 2,314 unique patient isolates of Moraxella catarrhalis were collected between October 1997 and June 2002 from 25 medical centers in 9 of the 10 Canadian provinces. Among the 7,566 H. influenzae isolates, 22.5% produced beta-lactamase, while 92.4% of the 2,314 M. catarrhalis isolates produced beta-lactamase. The incidence of beta-lactamase-producing H. influenzae isolates decreased significantly over the 5-year study period, from 24.2% in 1997-1998 to 18.6% in 2001-2002 (P < 0.01). The incidence of beta-lactamase-producing M. catarrhalis isolates did not change over the study period. The overall rates of resistance to amoxicillin and amoxicillin-clavulanate for H. influenzae were 19.3 and 0.1%, respectively. The rank order of cephalosporin activity based on the MICs at which 90% of isolates were inhibited (MIC(90)s) was cefotaxime > cefixime > cefuroxime > cefprozil > cefaclor. On the basis of the MICs, azithromycin was more active than clarithromycin (14-OH clarithromycin was not tested); however, on the basis of the NCCLS breakpoints, resistance rates were 2.1 and 1.6%, respectively. Rates of resistance to other agents were as follows: doxycycline, 1.5%; trimethoprim sulfamethoxazole, 14.2%; and chloramphenicol, 0.2%. All fluoroquinolones tested, including the investigational fluoroquinolones BMS284756 (garenoxacin) and ABT 492, displayed potent activities against H. influenzae, with MIC(90)s of < or = 0.03 microg/ml. The MIC(90)s of the investigational ketolides telithromycin and ABT-773 were 2 and 4 microg/ml, respectively, and the MIC(90) of the investigational glycylcycline GAR-936 (tigecycline) was 4 microg/ml. Among the M. catarrhalis isolates tested, the resistance rates derived by using the NCCLS breakpoint criteria for H. influenzae were <1% for all antibiotics tested except trimethoprim-sulfamethoxazole (1.5%). In summary, the incidence of beta-lactamase positive H. influenzae strains in Canada is decreasing (18.6% in 2001-2002), while the incidence of beta-lactamase-positive M. catarrhalis strains has remained constant (90.0% in 2001-2002). PMID- 12760862 TI - Treatment with a broad-spectrum cephalosporin versus piperacillin-tazobactam and the risk for isolation of broad-spectrum cephalosporin-resistant Enterobacter species. AB - Receipt of a broad-spectrum cephalosporin is a strong risk factor for isolation of broad-spectrum cephalosporin-resistant Enterobacter species, and yet the risk from other broad-spectrum beta-lactams hydrolyzed by group 1 beta-lactamases has not been well characterized. We compared the risk conferred by broad-spectrum cephalosporins to that conferred by piperacillin-tazobactam, alone or in combination with an aminoglycoside or a fluoroquinolone. A retrospective cohort was monitored from treatment onset until a broad-spectrum cephalosporin-resistant Enterobacter strain was isolated or the patient was discharged. There were 447 patients in the piperacillin-tazobactam group and 2,341 patients in the broad spectrum cephalosporin group. Groups were similar in age (mean, 62.5 years). The piperacillin-tazobactam group had a smaller percentage of men (32% versus 44%, P < 0.001) and a lower rate of intensive care unit stay (25% versus 38%, P < 0.001) but a higher rate of surgery (41% versus 26%, P < 0.001). Groups differed in the distribution of comorbidities. Resistant Enterobacter strains were isolated from 62 patients, 2% in each group (hazard ratio [RR] = 1.02 [P = 0.95]). In multivariable analysis, risk was similar among treatment groups (RR = 0.71 [P = 0.32]). Intensive care unit stay and surgery were associated with increased risk (RR = 4.53 [P < 0.001] and RR = 1.97 [P = 0.015], respectively), fluoroquinolones were protective (RR = 0.24 [P = 0.003]), and aminoglycosides did not affect risk (RR = 0.98 [P = 0.95]). The protective effect of fluoroquinolones against isolation of broad-spectrum cephalosporin-resistant Enterobacter spp. and the equivalence in risk associated with piperacillin-tazobactam and broad-spectrum cephalosporins may have important clinical and epidemiologic implications. PMID- 12760863 TI - Genetic and phenotypic variations of a resistant Pseudomonas aeruginosa epidemic clone. AB - From May 1997 to December 2001, a serotype O:6 multidrug-resistant strain of Pseudomonas aeruginosa colonized or infected 201 patients in the University Hospital of Besancon (France). The susceptibility profile of this epidemic clone to fluoroquinolones and aminoglycosides was relatively stable during the outbreak but showed important isolate-to-isolate variations (up to 64-fold) in the MICs of beta-lactams. Analysis of 18 genotypically related isolates selected on a quarterly basis demonstrated alterations in the two DNA topoisomerases II and IV (Thr83-->Ile in GyrA and Ser87-->Leu in ParC) and production of an ANT(2")-I enzyme. Although constitutively overproduced in these bacteria, the MexXY efflux system did not appear to contribute significantly to aminoglycoside resistance. beta-Lactam resistance was associated with derepression of intrinsic AmpC beta lactamase (with isolate-to-isolate variations of up to 58-fold) and sporadic deficiency in a 46-kDa protein identified as the carbapenem-selective porin OprD. Of the 18 isolates, 14 were also found to overproduce the efflux system MexAB OprM as a result of alteration of the repressor protein MexR (His107-->Pro). However, complementation experiments with the cloned mexR gene demonstrated that MexAB-OprM contributed only marginally to beta-lactam and fluoroquinolone resistance. Of the four isolates exhibiting wild-type MexAB-OprM expression despite the MexR alteration, two appeared to harbor secondary mutations in the mexA-mexR intergenic region and one harbored secondary mutations in the putative ribosome binding site located upstream of the mexAB oprM operon. In conclusion, this study shows that many mechanisms were involved in the multiresistance phenotype of this highly epidemic strain of P. aeruginosa. Our results also demonstrate that the clone sporadically underwent substantial genetic and phenotypic variations during the course of the outbreak, perhaps in relation to local or individual selective drug pressures. PMID- 12760864 TI - Antileishmanial activity of a linalool-rich essential oil from Croton cajucara. AB - The in vitro leishmanicidal effects of a linalool-rich essential oil from the leaves of Croton cajucara against Leishmania amazonensis were investigated. Morphological changes in L. amazonensis promastigotes treated with 15 ng of essential oil per ml were observed by transmission electron microscopy; leishmanial nuclear and kinetoplast chromatin destruction, followed by cell lysis, was observed within 1 h. Pretreatment of mouse peritoneal macrophages with 15 ng of essential oil per ml reduced by 50% the interaction between these macrophages and L. amazonensis, with a concomitant increase by 220% in the level of nitric oxide production by the infected macrophages. Treatment of preinfected macrophages with 15 ng of essential oil per ml reduced by 50% the interaction between these cells and the parasites, which led to a 60% increase in the amount of nitric oxide produced by the preinfected macrophages. These results provide new perspectives on the development of drugs with activities against Leishmania, as linalool-rich essential oil is a strikingly potent leishmanicidal plant extract (50% lethal doses, 8.3 ng/ml for promastigotes and 8.7 ng/ml for amastigotes) which inhibited the growth of L. amazonensis promastigotes at very low concentrations (MIC, 85.0 pg/ml) and which presented no cytotoxic effects against mammalian cells. PMID- 12760865 TI - In vitro activities of linezolid combined with other antimicrobial agents against Staphylococci, Enterococci, Pneumococci, and selected gram-negative organisms. AB - The activities of linezolid, an oxazolidinone antibacterial agent active against gram-positive organisms, alone and in combination with 35 antimicrobial agents were tested in vitro against methicillin-sensitive (n = 1 to 2 strains) and methicillin-resistant (n = 8 to 10) Staphylococcus aureus strains; vancomycin sensitive (n = 6) and vancomycin-resistant (n = 6 to 8) Enterococcus faecalis strains; vancomycin-sensitive (n = 5) and vancomycin-resistant (n = 6) Enterococcus faecium strains; penicillin-sensitive (n = 2 to 5), penicillin intermediate (n = 5 to 6), and penicillin-resistant (n = 5 to 6) Streptococcus pneumoniae strains; Escherichia coli (n = 6); and Klebsiella pneumoniae (n = 6). The fractional inhibitory concentration indices of linezolid in combination with other antimicrobial agents for the organisms tested were generated on checkerboard broth microdilution plates prepared by a semiautomated method. Of 1,380 organism-drug combinations, 1,369 (99.2%) combinations of linezolid with 28 antimicrobial drugs were indifferent, 9 combinations (0.65%) of linezolid with 6 drugs (amoxicillin, erythromycin, imipenem, sparfloxacin, teicoplanin, and tetracycline) were synergistic, and 2 combinations (0.15%) of linezolid with 2 drugs (ofloxacin and sparfloxacin) were antagonistic. Overall, the in vitro data demonstrated that linezolid combined with other antimicrobial agents primarily produces an indifferent response, with infrequent occurrences of synergism and antagonism. PMID- 12760866 TI - Experimental study of LY333328 (oritavancin), alone and in combination, in therapy of cephalosporin-resistant pneumococcal meningitis. AB - Using a rabbit model of meningitis, we sought to determine the efficacy of LY333328, a semisynthetic glycopeptide, in the treatment of cephalosporin resistant pneumococcal meningitis. LY333328 was administered at a dose of 10 mg/kg of body weight/day, alone and in combination with ceftriaxone at 100 mg/kg/day with or without dexamethasone at 0.25 mg/kg/day. The therapeutic groups were treated with LY333328 with or without dexamethasone and LY333328-ceftriaxone with or without dexamethasone. Rabbits were inoculated with a cephalosporin resistant pneumococcal strain (ceftriaxone MIC, 2 microg/ml; penicillin MIC, 4 microg/ml; LY333328 MIC, 0.008 microg/ml) and were treated over a 26-h period beginning 18 h after inoculation. The bacterial counts in cerebrospinal fluid (CSF), the white blood cell count, the lactic acid concentration, the CSF LY333328 concentration, and bactericidal and bacteriostatic activities were determined at different time points. In vitro, LY333328 was highly bactericidal and its use in combination with ceftriaxone at one-half the MIC was synergistic. In the rabbit model, LY333328 alone was an excellent treatment for cephalosporin resistant pneumococcal meningitis, with a rapid decrease in colony counts and no therapeutic failures. The use of LY333328 in combination with ceftriaxone improved the activity of LY333328, but no synergistic effect was observed. The combination of LY333328 with dexamethasone was also rapidly bactericidal, but two therapeutic failures were observed. The combination of LY333328 with ceftriaxone and dexamethasone was effective, without therapeutic failures. PMID- 12760867 TI - Ribavirin and alpha interferon enhance death receptor-mediated apoptosis and caspase activation in human hepatoma cells. AB - The molecular mechanisms underlying the clinical effects of alpha interferon (IFN) and ribavirin are not understood. Elimination of infected cells occurs in part by cytotoxic T lymphocytes (CTLs) expressing CD95 ligand and thereby attacking target cells which are positive for the death receptor CD95. Since many viruses have evolved mechanisms to inhibit apoptosis, the opposite, namely, promotion of apoptosis, could be a strategy to strengthen the host antiviral response. In the present study, we have asked whether the antiviral substances IFN and ribavirin could support CD95-mediated apoptosis by interfering with the activation of caspases, a family of proteases known for their essential role in apoptosis. HepG2 cells, stimulated with the agonistic anti-CD95 antibody, served as a minimal model to mimic the CD95 stimulation occurring during a CTL attack of target cells in vivo. Apoptosis was quantitated by flow cytometric detection of hypodiploid nuclei. Caspase activity was measured by cytofluorometry, immunocytochemistry, and immunoblot analysis. IFN and ribavirin sensitized HepG2 cells for CD95-mediated apoptosis. This effect was correlated with an increase in CD95-mediated caspase activation and enhanced cleavage of the caspase substrate poly(ADP-ribose) polymerase. Furthermore, the positive effect on CD95-mediated caspase activation by IFN and ribavirin was confirmed by immunocytochemistry for activated caspase-3 and by immunoblot detection of activated caspase-3, caspase 7, and caspase-8. Our data demonstrate that the antiviral substances IFN and ribavirin are able to sensitize for CD95-mediated apoptosis. IFN and ribavirin also enhance CD95-mediated caspase activation, which might in part be responsible for the apoptosis-promoting effect of these antiviral compounds. PMID- 12760868 TI - Antiviral activity and pharmacokinetics of 1-(2,3-dideoxy-2-fluoro-beta-L glyceropent-2-enofuranosyl)cytosine. AB - 1-(2,3-Dideoxy-2-fluoro-beta-L-glyceropent-2-enofuranosyl)cytosine (L-2'-Fd4C) is an L-nucleoside analogue with both anti-human immunodeficiency virus (HIV) and anti-hepatitis B virus (HBV) activity with median effective concentrations of 0.12 microM in peripheral blood mononuclear cells and 0.002 microM in HepG2 2.2.15 cells, respectively. The purpose of this study was to examine the antihepadnavirus potency and pharmacokinetics of L-2'-Fd4C in vivo. HBV transgenic mice treated intraperitoneally with L-2'-Fd4C showed a reduction of HBV levels in their blood comparable to that produced by lamivudine. The pharmacokinetics of L-2'-Fd4C in rhesus monkeys was evaluated after intravenous and oral administration. The concentrations in plasma declined in a biexponential manner after intravenous administration, with a long terminal-phase half-life of 5.02 h. The steady-state volumes of distribution and systemic clearance were 1.09 liter x kg(-1) and 0.25 liter x h(-1) x kg(-1), respectively, with a renal clearance of 0.16 liter x h(-1) x kg(-1). The oral bioavailability was approximately 44%. About 53% of the compound administered intravenously and 19% of that administered orally were recovered unchanged in the urine within the 24-h urine collection period, and no other metabolite was detected. The compound penetrated the central nervous system at concentrations that exceeded the median effective antiviral concentration against HIV in cell cultures. Based upon these observations, further testing to develop this agent for treatment of HIV and HBV infections is warranted. PMID- 12760869 TI - Indinavir, efavirenz, and abacavir pharmacokinetics in human immunodeficiency virus-infected subjects. AB - Adult AIDS Clinical Trials Group (AACTG) Protocol 886 examined the dispositions of indinavir, efavirenz, and abacavir in human immunodeficiency virus-infected subjects who received indinavir at 1,000 mg every 8 h (q8h) and efavirenz at 600 mg q24h or indinavir at 1,200 mg and efavirenz at 300 mg q12h with or without abacavir 300 at mg q12h. Thirty-six subjects participated. The median minimum concentration in plasma (C(min)) for indinavir administered at 1,200 mg q12h was 88.1 nM (interquartile range [IR], 61.7 to 116.5 nM), whereas the median C(min) for indinavir administered at 1,000 mg q8h was 139.3 nM (IR, 68.8 to 308.7 nM) (P = 0.19). Compared to the minimum C(min) range for wild-type virus (80 to 120 ng/ml) estimated by the AACTG Adult Pharmacology Committee, the C(min) for indinavir administered at 1,200 mg q12h (54 ng/ml) is inadequate. The apparent oral clearance (CL/F) (P = 0.28), apparent volume of distribution at steady state (V(ss)/F) (P = 0.25), and half-life (t(1/2)) (P = 0.80) of indinavir did not differ between regimens. The levels of efavirenz exposure were similar between regimens. For efavirenz administered at 600 mg q24h and 300 mg q12h, the median maximum concentrations in plasma (C(max)s) were 8,968 nM (IR, 5,784 to 11,768 nM) and 8,317 nM (6,587 to 10,239 nM), respectively (P = 0.66), and the C(min)s were 4,289 nM (IR, 2,462 to 5,904 nM) and 4,757 nM (IR, 3,088 to 6,644 nM), respectively (P = 0.29). Efavirenz pharmacokinetic parameters such as CL/F (P = 0.62), V(ss)/F (P = 0.33), and t(1/2) (P = 0.37) were similar regardless of the dosing regimen. The median C(max), C(min), CL/F, V(ss)/F, and t(1/2) for abacavir were 6,852 nM (IR, 5,702 to 7,532), 21.0 nM (IR, 21.0 to 87.5), 43.7 liters/h (IR, 37.9 to 55.2), 153.9 liters (IR, 79.6 to 164.4), and 2.0 h (IR, 1.8 to 2.8), respectively. In summary, when indinavir was given with efavirenz, the trough concentration of indinavir after administration of 1,200 mg q12h was inadequate. Abacavir did not influence the pharmacokinetics or exposure parameters of either indinavir or efavirenz. The levels of efavirenz exposure were similar in subjects receiving efavirenz q12h or q24h. PMID- 12760871 TI - Activities of ertapenem, a new long-acting carbapenem, against penicillin sensitive or -resistant pneumococci in experimental meningitis. AB - The penetration of ertapenem, a new carbapenem with a long half-life, reached 7.1 and 2.4% into inflamed and noninflamed meninges, respectively. Ertapenem had excellent antibacterial activity in the treatment of experimental meningitis due to penicillin-sensitive and -resistant pneumococci, leading to a decrease of 0.69 +/- 0.17 and 0.59 +/- 0.22 log(10) CFU/ml x h, respectively, in the viable cell counts in the cerebrospinal fluid. The efficacy of ertapenem was comparable to that of standard regimens (ceftriaxone monotherapy against the penicillin sensitive strain and ceftriaxone combined with vancomycin against the penicillin resistant strain). In vitro, ertapenem in concentrations above the MIC was highly bactericidal against both strains. Even against a penicillin- and quinolone resistant mutant, ertapenem had similar bactericidal activity in vitro. PMID- 12760870 TI - Generation of stable cell lines expressing Lamivudine-resistant hepatitis B virus for antiviral-compound screening. AB - Lamivudine [beta-L-(-)-2',3'-dideoxy-3'-thiacytidine] is a potent inhibitor of hepadnavirus replication and is used both to treat chronic hepatitis B virus (HBV) infections and to prevent reinfection of transplanted livers. Unfortunately, lamivudine-resistant HBV variants do arise during prolonged therapy, indicating a need for additional antiviral drugs. Replication-competent HBV constructs containing the reverse transcriptase domain L180M/M204V and M204I (rtL180M/M204V and rtM204I) mutations associated with lamivudine resistance were used to produce stable cell lines that express the resistant virus. These cell lines contain stable integrations of HBV sequences and produce both intracellular and extracellular virus. HBV produced by these cell lines was shown to have a marked decrease in sensitivity to lamivudine, with 450- and 3,000-fold shifts in the 50% inhibitory concentrations for the rtM204I and rtL180M/M204V viruses, respectively, compared to that for the wild-type virus. Drug assays indicated that the lamivudine-resistant virus exhibited reduced sensitivity to penciclovir [9-(4-hydroxy-3-hydroxymethyl-but-1-yl) guanine] but was still inhibited by the nucleoside analogues CDG (carbocyclic 2'-deoxyguanosine) and abacavir ([1S,4R]-4 [2-amino-6-(cyclopropylamino)-9H-purin-9-yl]-2-cyclopentene-1-methanol). Screening for antiviral compounds active against the lamivudine-resistant HBV can now be done with relative ease. PMID- 12760872 TI - Efficacy of albaconazole (UR-9825) in treatment of disseminated Scedosporium prolificans infection in rabbits. AB - There are no effective therapeutics for treating invasive Scedosporium prolificans infections. Doses of 15, 25, and 50 mg/kg of body weight/day for the new triazole albaconazole (ABC) were evaluated in an immunocompetent rabbit model of systemic infection with this mold. Treatments were begun 1 day after challenge and given for 10 days. ABC at any dose was more effective than amphotericin B (AMB) at 0.8 mg/kg/day at clearing S. prolificans from tissue (P < 0.007). The percentages of survival at 25 mg of ABC/kg/day were similar to those obtained with AMB. Rabbits showed 100% survival when they were treated with 50 mg of ABC per kg (P < 0.0001 versus control group), and only this dosage was able to reduce tissue burden significantly in the five organs studied, i.e., spleen, kidneys, liver, lungs, and brain. PMID- 12760873 TI - Pharmacokinetic-pharmacodynamic modeling of the electroencephalogram effect of norfloxacin in rats. AB - A previously developed pharmacokinetic-pharmacodynamic (PK-PD) modeling approach was used to investigate the epileptogenic activity of norfloxacin as a representative antibiotic with concentration-dependent antimicrobial activity. Rats received an intravenous infusion of norfloxacin at a rate of 5 mg kg of body weight(-1) min(-1) over 30 min. Blood samples were collected for drug assay, and an electroencephalogram (EEG) was recorded during infusion and postinfusion. An important delay was observed between concentrations of norfloxacin in plasma and the EEG effect. Indirect effect models failed to describe these data, which were successfully fitted by using an effect compartment model with a spline function to describe the relationship between effect and concentration at the effect site, as previously observed with imipenem. The robustness of the PK-PD model was then assessed by keeping the dose constant but increasing the duration of infusion to 120 and 240 min. Although this was accompanied by PK modifications, PD parameters did not vary significantly, and the PK-PD model still applied. In conclusion, the successful PK-PD modeling of the norfloxacin EEG effect in rats should be considered to predict and reduce the epileptogenic risk associated with this antibiotic as a representative fluoroquinolone (E. Fuseau and L. B. Sheiner, Clin. Pharmacol. Ther. 35:733-741, 1984). PMID- 12760874 TI - High prevalence of antimicrobial resistance in rapidly growing mycobacteria in Taiwan. AB - An increasing number of clinical isolations of rapidly growing mycobacteria (RGM) at the National Taiwan University Hospital were noted from 1992 to 2001. Broth microdilution MICs of 15 antimicrobial agents were determined for 200 clinical isolates of RGM, including the Mycobacterium fortuitum group (69 isolates), M. chelonae (39 isolates), and M. abscessus (92 isolates). Our results showed that the resistance rates of these isolates to the currently available agents were remarkably high. Amikacin was active against nearly all RGM isolates. Clarithromycin was usually active against M. abscessus (79% susceptibility) and the M. fortuitum group (65% susceptibility). The majority of M. fortuitum group isolates were susceptible to ciprofloxacin (62%) and imipenem (61%). The susceptibilities to other conventional anti-RGM agents of these isolates were poor but differed markedly by species. The newer fluoroquinolones (levofloxacin, moxifloxacin, and gatifloxacin) and meropenem showed better in vitro activities against the M. fortuitum group isolates than against the other two species of RGM. Linezolid had fairly good activity against these RGM isolates, particularly against M. chelonae isolates (82% susceptible). Telithromycin had poor activity against these RGM isolates (the MICs at which 50% of the isolates tested are inhibited [MIC(50)s] were 32 to 64 microg/ml, and the MIC(90)s were >64 microg/ml). PMID- 12760876 TI - In vitro activities of dalbavancin and nine comparator agents against anaerobic gram-positive species and corynebacteria. AB - Dalbavancin is a novel semisynthetic glycopeptide with enhanced activity against gram-positive species. Its comparative in vitro activities and those of nine comparator agents, including daptomycin, vancomycin, linezolid, and quinupristin dalfopristin, against 290 recent gram-positive clinical isolates strains, as determined by the NCCLS agar dilution method, were studied. The MICs of dalbavancin at which 90% of various isolates tested were inhibited were as follows: Actinomyces spp., 0.5 microg/ml; Clostridium clostridioforme, 8 microg/ml; C. difficile, 0.25 microg/ml; C. innocuum, 0.25 microg/ml; C. perfringens, 0.125 microg/ml; C. ramosum, 1 microg/ml; Eubacterium spp., 1 microg/ml; Lactobacillus spp., >32 microg/ml, Propionibacterium spp., 0.5 microg/ml; and Peptostreptococcus spp., 0.25 microg/ml. Dalbavancin was 1 to 3 dilutions more active than vancomycin against most strains. Dalbavancin exhibited excellent activity against gram-positive strains tested and warrants clinical evaluation. PMID- 12760875 TI - In vitro activities of telithromycin and 10 oral agents against aerobic and anaerobic pathogens isolated from antral puncture specimens from patients with sinusitis. AB - A study of the comparative in vitro activity of telithromycin, a new ketolide, against 155 aerobic and 171 anaerobic antral sinus puncture isolates showed it to be active against a broad range of sinus pathogens. All pneumococci, including erythromycin-resistant strains, were susceptible to telithromycin at < or = 0.5 microg/ml; all Haemophilus influenzae and Eikenella corrodens strains were inhibited by < or = 4 microg of telithromycin/ml; all Moraxella spp. and beta lactamase-producing Prevotella species strains were inhibited by < or = 0.25 and 0.5 microg of telithromycin/ml, respectively. Among all anaerobes tested, 94% (160 of 171 strains) were susceptible to < or = 4 microg of telithromycin/ml; however, 8 of 17 (47%) Fusobacterium strains, 2 Veillonella strains, and 1 Peptostreptococcus micros strain required > 4 microg of telithromycin/ml for inhibition. Telithromycin may offer a therapeutic alternative for sinus infections, including those due to erythromycin-resistant pneumococci. PMID- 12760877 TI - Chlamydia pneumoniae resists antibiotics in lymphocytes. AB - Chlamydia pneumoniae infection of lymphocytes in blood has been well documented, and it is apparent that control of this pathogen in these cells may be critical in the development of chronic inflammatory diseases associated with infection by this bacterium. The activity of antibiotics against C. pneumoniae in lymphocytes was assessed in this study by utilizing an in vitro infection model with lymphoid cells. The results obtained indicated that although all of the antibiotics tested showed remarkable activity against bacterial growth in epithelial cells, C. pneumoniae in lymphocytes was less susceptible to antibiotics than was bacterial growth in epithelial cells, which are widely used for the evaluation of anti-C. pneumoniae antibiotics. PMID- 12760878 TI - Lack of vancomycin tolerance in Streptococcus pneumoniae strains isolated in Barcelona, Spain, from 1999 to 2001. AB - To evaluate the incidence of vancomycin tolerance among Streptococcus pneumoniae isolates, we performed killing curve studies with 633 isolates. The penicillin MIC was > or = 0.12 mg/liter for 481 (76%) of the isolates. All strains were susceptible to vancomycin. Killing curve studies were performed with a vancomycin concentration of 2.5 mg/liter. The Tupelo strain was used for quality control. No vancomycin-tolerant strain was detected. PMID- 12760879 TI - RNA III inhibiting peptide inhibits in vivo biofilm formation by drug-resistant Staphylococcus aureus. AB - Staphylococcus aureus is a prevalent cause of bacterial infections associated with indwelling medical devices. RNA III inhibiting peptide (RIP) is known to inhibit S. aureus pathogenesis by disrupting quorum-sensing mechanisms. RIP was tested in the present study for its ability to inhibit S. aureus biofilm formation in a rat Dacron graft model. The activity of RIP was synergistic with those of antibiotics for the complete prevention of drug-resistant S. aureus infections. PMID- 12760880 TI - Linezolid and vancomycin, alone and in combination with rifampin, compared with moxifloxacin against a multidrug-resistant and a vancomycin-tolerant Streptococcus pneumoniae strain in an in vitro pharmacodynamic model. AB - Potential alternatives, including linezolid, adjunctive rifampin, and moxifloxacin, were evaluated against vancomycin-tolerant (P9802-020) and vancomycin-susceptible clinical isolates of Streptococcus pneumoniae in an in vitro pharmacodynamic model. Vancomycin exhibited maximal killing of 2-log(10) CFU/ml against P9802-020. Linezolid, moxifloxacin, and linezolid plus rifampin exhibited 99.9% killing against both isolates. These alternatives should be considered for further evaluation against vancomycin-tolerant S. pneumoniae. PMID- 12760881 TI - Eradication of resident bacteria of normal human skin by antimicrobial ointment. AB - The application of a topical triple-antibiotic ointment (containing neomycin, polymyxin, and bacitracin) eradicated resident bacteria through 25 layers of the stratum corneum and prevented repopulation of bacteria overnight but not at 1 week. Through 15 layers, mupirocin had some effect, whereas a double-antibiotic ointment had none. The reservoir of resident bacteria in the sebaceous glands is not affected by a topical antibiotic. PMID- 12760882 TI - Stability and antibacterial activity of cefepime during continuous infusion. AB - The stability of cefepime during simulated continuous infusion was determined with a motorized portable infusion pump worn over a period of 24 to 36 h. Susceptibility testing on cefepime solutions over time indicates that the degradation products do not exhibit antibacterial activity. Cefepime stability at 24 h following continuous infusion was 94.3% +/- 1.0%, which supports the use of continuous infusion. PMID- 12760883 TI - Killing activity of micafungin against Aspergillus fumigatus hyphae assessed by specific fluorescent staining for cell viability. AB - We studied the anti-Aspergillus activity of micafungin by using two fluorescent dyes to detect cell viability. Micafungin induced flattened hyphae, caused by the bursting of cells, which had lost their viability. Micafungin has killing activity against actively growing hyphae, even though it is not fungicidal against the whole burden of Aspergillus fumigatus. PMID- 12760884 TI - Risk factors for fecal quinolone-resistant Escherichia coli in Mexican children. AB - We determined the prevalence of, and risk factors for, fecal quinolone-resistant Escherichia coli (QREC) in 324 children from Yucatan, Mexico. QREC was higher in children with recent Salmonella infection (100%) than in children with diarrhea (61%) or healthy children (54%) (P = 0.007). Multivariate analysis identified recent hospitalization of a family member (P = 0.011, odds ratio [OR] = 5.1) and carriage of Salmonella (P = 0.004, OR = 3.7) as independent risk factors for QREC. PMID- 12760885 TI - Evolution of antibiotic resistance in Salmonella enterica serovar typhimurium strains isolated in the Czech Republic between 1984 and 2002. AB - In a collection of 66 Salmonella enterica serovar Typhimurium strains isolated between 1984 and 2002 in the Czech Republic, genes coding for antibiotic resistance were determined by using specific PCRs. We found that the pentadrug resistant ACSSuT clone first appeared in the Czech Republic in 1990. A new variant of the aadA gene designated aadA21 is described, the 5' end of which was identical to aadA2 and the 3' end of which was identical to aadA1. PMID- 12760886 TI - Salmonella enterica serovar typhi strains isolated in Korea containing a multidrug resistance class 1 integron. AB - Six strains of Salmonella enterica serovar Typhi which were resistant to ampicillin, chloramphenicol, trimethoprim-sulfamethoxazole, streptomycin, tetracycline, and gentamicin were isolated in Korea. This multidrug resistance was transferred by a conjugative plasmid of about 50 kb. The plasmid harbored a class 1 integron, which included six resistance genes, aacA4b, catB8, aadA1, dfrA1, aac(6')-IIa, and the novel blaP2, in that order. All of the isolates showed the same-size plasmids and the same ribotyping patterns, which suggests a clonal spread of these multidrug-resistant isolates. PMID- 12760887 TI - Molecular characterization of isoniazid-resistant Mycobacterium tuberculosis clinical isolates in Lithuania. AB - Mutations at codon 315 of the katG gene were detected in 312 of 364 (85.7%) isoniazid-resistant Mycobacterium tuberculosis isolates. Seven of 52 (13.5%) isoniazid-resistant isolates with the wild-type Ser315 codon and 10 of 52 (19.2%) isoniazid-resistant isolates with a mutated katG allele had mutation -15C-->T in the promoter of the mabA-inhA operon. PMID- 12760888 TI - Comparative roles of the cell wall and cell membrane in limiting uptake of xenobiotic molecules by Saccharomyces cerevisiae. AB - Using reversible electropermeabilization of cells and spheroplasts, we show that the cell wall and plasma membrane partly account for bleomycin resistance by acting as two independent barriers. We also report on the presence of a membrane protein that may be responsible for bleomycin internalization and toxicity in Saccharomyces cerevisiae. PMID- 12760889 TI - Vancomycin penetration of uninfected pleural fluid exudate after continuous or intermittent infusion. AB - Blood and pleural exudate samples were obtained from 16 patients receiving intermittent or continuous infusions of vancomycin after lung surgery. The areas under the concentration-time curves for blood and pleural exudates were identical for both administration schedules, while continuous infusion allowed the concentrations in pleural exudates to be more sustained (mean concentration, 12 mg/liter). PMID- 12760890 TI - Influence of reduced susceptibility to glycopeptides on activities of vancomycin and teicoplanin against Staphylococcus aureus in experimental endocarditis. AB - The influence of reduced susceptibilities to glycopeptides on the activities of vancomycin and teicoplanin against an isogenic pair of clinical Staphylococcus aureus strains in experimental endocarditis was investigated. While vancomycin was similarly active against both strains, teicoplanin was approximately 100-fold less active against the resistant strain and selected for the emergence of more resistant subpopulations. PMID- 12760891 TI - Alpha interferon augments cidofovir's antiviral and antiproliferative activities. AB - The antiviral and antiproliferative activities of alpha 2a interferon (IFN-alpha 2a) and cidofovir in human papillomavirus type 16 (HPV-16)-transformed keratinocytes were evaluated. The compounds in combination were more effective than comparable levels of either drug alone. Evaluation of effective drug ratios revealed a synergistic cooperation between IFN-alpha 2a and cidofovir in inhibiting the proliferation of HPV-infected cells. PMID- 12760892 TI - Antimicrobial resistance of Campylobacter jejuni subsp. jejuni strains isolated from humans in 1998 to 2001 in Montreal, Canada. AB - The rates of resistance of 51 to 72 human strains of Campylobacter jejuni subsp. jejuni isolated annually from 1998 to 2001 in Montreal, Quebec, Canada, varied from 1 to 12% for erythromycin, 43 to 68% for tetracycline, and 10 to 47% for ciprofloxacin. In the last years of the study, there was a significant increase in the rate of resistance to ciprofloxacin (P = 0.00003) but not in the rate of resistance to erythromycin (P = 0.056) or tetracycline (P = 0.095) compared to the rate obtained in the first years. All 51 C. jejuni strains isolated in 2001 were susceptible to gentamicin, amoxicillin-clavulanic acid, imipenem, and meropenem. From 1999 to 2001, 74 strains of C. jejuni acquired abroad were significantly more resistant to ciprofloxacin than 109 strains of C. jejuni acquired locally (66 versus 9%, P < 0.00001) but were not significantly more resistant to erythromycin (1 versus 6%, P = 0.15) or to tetracycline (55 versus 58%, P = 0.87). PMID- 12760893 TI - Emergence of resistance in normal human aerobic commensal flora during telithromycin and amoxicillin-clavulanic acid treatments. AB - Mean fecal global yeast counts increased similarly during 7 days of treatment with telithromycin (800 mg once daily) or amoxicillin-clavulanic acid (amoxiclav) (1 g of amoxicillin and 125 mg of clavulanic acid 3 times daily) in human volunteers and decreased slowly thereafter. On skin, coagulase-negative staphylococci of decreased susceptibility (DS) to telithromycin increased in the telithromycin group, whereas those with DS to methicillin increased in the amoxiclav group. A similar antibiotic-related shift towards homologous DS was observed for oral nongroupable streptococci (NGS), but in addition, the prevalence of NGS resistant to both classes of antibiotics was significantly greater in the amoxiclav group at days 8 (P < 0.01) and 45 (P < 0.015). PMID- 12760894 TI - Resistance to autolysis in vancomycin-selected Staphylococcus aureus isolates precedes vancomycin-intermediate resistance. AB - Four clinical U.S. glycopeptide intermediate resistant Staphylococcus aureus (GISA) isolates were resistant to Triton X-100-induced autolysis. Similar resistance was demonstrated in an isolate obtained after a single passage of a susceptible clinical isolate in low-level vancomycin. Strains with the vancomycin induced Triton X-100 resistance phenotype produced active murein hydrolases but were resistant to lysis by murein hydrolases. PMID- 12760895 TI - Biochemical characterization of beta-lactamases Bla1 and Bla2 from Bacillus anthracis. AB - The Sterne and Ames strains of Bacillus anthracis carry chromosomal genes bla1 and bla2, which confer beta-lactam resistance when expressed in Escherichia coli. MIC measurements and steady-state kinetic analyses indicate that Bla1 possesses penicillinase activity while Bla2 possesses penicillinase, cephalosporinase, and carbapenem-hydrolyzing activities. PMID- 12760896 TI - Prodrugs of cephalosporin RWJ-333441 (MC-04,546) with improved aqueous solubility. AB - To improve the aqueous solubility of anti-methicillin-resistant Staphylococcus aureus cephalosporin RWJ-333441 for parenteral administration, acyl derivatives of the C-3 primary amino group were prepared and evaluated for solubility, cleavage in serum in vitro, and conversion to RWJ-333441 in vivo. Improved solubility at physiologic pH values and release of RWJ-333441 in vitro and in vivo were observed for several prodrugs, including the aspartate derivative RWJ 333442. PMID- 12760897 TI - Mechanism of action of 4-phenoxyphenoxyethyl thiocyanate (WC-9) against Trypanosoma cruzi, the causative agent of Chagas' disease. AB - We investigated the molecular basis of the activity of 4-phenoxyphenoxyethyl thiocyanate (WC-9) against Trypanosoma cruzi, the etiological agent of Chagas' disease. We found that growth inhibition of T. cruzi epimastigotes induced by this compound was associated with a reduction in the content of the parasite's endogenous sterols due to a specific blockade of their de novo synthesis at the level of squalene synthase. PMID- 12760899 TI - Detection of CMY-2, CTX-M-14, and SHV-12 beta-lactamases in Escherichia coli fecal-sample isolates from healthy chickens. AB - Genes encoding the CMY-2, CTX-M-14, and SHV-12 beta-lactamases were detected in three of five Escherichia coli isolates from fecal samples from healthy chickens which showed resistance or diminished susceptibility to extended-spectrum cephalosporins. A -42 mutation at the promoter region of the ampC gene was detected in the other two isolates. PMID- 12760898 TI - Inhibitors of pantothenate kinase: novel antibiotics for staphylococcal infections. AB - Pantothenate kinase (CoaA) catalyzes the first step of the coenzyme A biosynthetic pathway. Here we report the identification of the Staphylococcus aureus coaA gene and characterization of the enzyme. We have also identified a series of low-molecular-weight compounds which are effective inhibitors of S. aureus CoaA. PMID- 12760900 TI - Multiple resistance mechanisms in fluoroquinolone-resistant Salmonella isolates from Germany. PMID- 12760901 TI - New antimicrobial agents approved by the U.S. Food and Drug Administration in 2002 and new indications for previously approved agents. PMID- 12760902 TI - Regulation of IL-11 expression in intestinal myofibroblasts: role of c-Jun AP-1- and MAPK-dependent pathways. AB - IL-11 inhibits the activation of NF-kappaB and induces the Th2 polarization of CD4+ T cells. The clinical utility of IL-11 is being investigated in Crohn's disease. However, physiological secretion of IL-11 in the intestine remains unclear. In this study, we investigated IL-11 secretion in human intestinal subepithelial myofibroblasts (SEMFs). Intestinal SEMFs were isolated from the human colonic mucosa. IL-11 secretion and mRNA expression were determined by ELISA and Northern blot analysis. The activating protein (AP)-1-DNA binding activity was evaluated by EMSA. IL-11 secretion was induced by IL-1beta and transforming growth factor (TGF)-beta1. These were also observed at the mRNA level. The EMSAs demonstrated that both IL-1beta and TGF-beta1 induced AP-1 activation within 2 h after stimulation, and a blockade of AP-1 activation by the recombinant adenovirus containing a dominant negative c-Jun markedly reduced the IL-1beta- and TGF-beta1-induced IL-11 mRNA expression. IL-1beta and TGF-beta1 induced an activation of ERK p42/44 and p38 MAP kinases, and the MAP kinase inhibitors (SB-202190, PD-98059, and U-0216) significantly reduced the IL-1beta- and TGF-beta1-induced IL-11 secretion. The upregulation of IL-11 mRNA by IL-1beta and TGF-beta1 was also mediated by a p38 MAP kinase-mediated mRNA stabilization. The combination of IL-1beta and TGF-beta1 additively enhanced IL-11 secretion. Intestinal SEMFs secreted IL-11 in response to IL-1beta- and TGF-beta1. Mucosal IL-11 secretion might be important as an anti-inflammatory response in the pathogenesis of intestinal inflammation. PMID- 12760903 TI - Influence of duodenal acidification on the sensorimotor function of the proximal stomach in humans. AB - Decreased acid clearance and increased exposure to acid of the duodenum have been reported in a subset of functional dyspepsia patients. However, the mechanism by which increased duodenal acid exposure may affect symptoms is unclear. The aim of the present study was to investigate the effects of duodenal acidification on proximal gastric tone and mechanosensitivity in humans. An infusion tube with a pH electrode attached was positioned in the second part of the duodenum, and a barostat bag was located in the gastric fundus. In 12 healthy subjects, fundic tone and sensitivity to distensions were assessed before and during duodenal infusion of 0.1 N hydrochloric acid or saline in a randomized, double-blind design. In 10 healthy subjects, meal-induced accommodation was measured during duodenal infusion of acid or saline. Acid infusion in the duodenum significantly increased fundic compliance and decreased fasting fundic tone. This was accompanied by a significant decrease in the pressures and the corresponding wall tensions at the thresholds for discomfort. During infusion of acid, significantly higher perception and symptom scores were obtained for the same distending pressures. The meal-induced fundic relaxation was significantly smaller during acid infusion compared with saline infusion. In conclusion, duodenal acidification induces proximal gastric relaxation, increases sensitivity to gastric distension, and inhibits gastric accommodation to a meal. Through these mechanisms, increased duodenal acid exposure may be involved in the pathogenesis of dyspeptic symptoms. PMID- 12760904 TI - Copper deficiency increases iron absorption in the rat. AB - Release of iron from enterocytes and hepatocytes is thought to require the copper dependent ferroxidase activity of hephaestin (Hp) and ceruloplasmin (Cp), respectively. In swine, copper deficiency (CD) impairs iron absorption, but whether this occurs in rats is unclear. By feeding a diet deficient in copper, CD was produced, as evidenced by the loss of copper-dependent plasma ferroxidase I activity, and in enterocytes, CD reduced copper levels and copper-dependent oxidase activity. Hematocrit was reduced, and liver iron was doubled. CD reduced duodenal mucosal iron and ferritin, whereas CD increased iron absorption. Duodenal mucosal DMT1-IRE and ferroportin1 expression remained constant with CD. When absorption in CD rats was compared with that seen normally and in iron deficient anemic animals, strong correlations were found among mucosal iron, ferritin, and iron absorption, suggesting that the level of iron absorption was appropriate given that the erythroid and stores stimulators of iron absorption are opposed in CD. Because CD reduced the activity of Cp, as evidenced by copper dependent plasma ferroxidase I activity and hepatocyte iron accumulation, but iron absorption increased, it is unlikely that the ferroxidase activity of Hp is important and suggests another function for this protein in the export of iron from the enterocyte during iron absorption. Also, the copper-dependent ferroxidase activity of Cp does not appear important for iron efflux from macrophages, because Kupffer cells of the liver and nonheme iron levels of the spleen were normal during copper deficiency, suggesting another role for Cp in these cells. PMID- 12760905 TI - Glucose regulates the cortical actin network through modulation of Cdc42 cycling to stimulate insulin secretion. AB - Glucose-stimulated insulin granule exocytosis in pancreatic beta-cells involves cortical actin remodeling that results in the transient disruption of the interaction between polymerized actin with the plasma membrane t-SNARE (target membrane soluble N-ethylmaleimide-sensitive factor attachment protein receptor) complex. To examine the mechanism underlying the initiation of cortical actin remodeling, we have used the actin nucleating/stabilizing agent jasplakinolide to show that remodeling is initiated at a step proximal to the ATP-sensitive K+ channels in the stimulus-secretion pathway. Confocal immunofluorescent microscopy revealed that cortical actin remodeling was required for glucose-stimulated insulin secretion. Furthermore, glucose was found to mediate the endogenous activation state of the Rho family GTPase Cdc42, a positive proximal effector of actin polymerization, resulting in a net decrease of Cdc42-GTP within 5 min of stimulation. Intriguingly, glucose stimulation resulted in the rapid and reversible glucosylation of Cdc42, suggesting that glucose inactivated Cdc42 by selective glucosylation to induce cortical actin rearrangement. Moreover, expression of the constitutively active form of Cdc42 (Q61L) inhibited glucose stimulated insulin secretion, whereas the dominant negative form (T17N) was without effect, suggesting that glucose-stimulated insulin secretion requires Cdc42 cycling to the GDP-bound state. In contrast, KCl-stimulated insulin secretion was unaffected by the expression of dominant negative or constitutively active Cdc42 and ceased to modulate endogenous Cdc42 activation, consistent with glucose-dependent cortical actin remodeling. These findings reveal that glucose regulates the cortical actin network through modulation of Cdc42 cycling to induce insulin secretion in pancreatic beta-cells. PMID- 12760906 TI - The effect of changes in ambient oxygen concentration on the bioelectric properties of middle ear mucosa. AB - The purpose of the present study was to compare the effect of 24 h of exposure to 7% O2 (normal middle ear physiological conditions) vs. 21% O2 (found in the middle ear after ventilation tube placement) on transepithelial Na+ absorption and Cl- secretion in cultured gerbil middle ear epithelial cell monolayers. Although no difference in apical Na+ absorption was identified, the UTP-induced stimulation of apical Cl- secretion in the presence of apical Na+ channel blockade with amiloride was significantly enhanced after exposure to 21% O2 compared with 7% O2 exposure. In the presence of a calcium-activated Cl- channel inhibitor, DIDS, UTP-induced stimulation of Cl- secretion after 21% O2 exposure was decreased, suggesting a role for calcium-activated Cl- channels in middle ear Cl- secretion in response to relative hyperoxia. PMID- 12760908 TI - "My word, what is that?": Hounsfield and the triumph of clinical research. PMID- 12760907 TI - LPP, a LIM protein highly expressed in smooth muscle. AB - An 80-kDa protein, prominently expressed in smooth muscle, was microsequenced and identified as LPP, the product of the lipoma-preferred partner gene (Petit MMR, Mols R, Schoenmakers EFPM, Mandahl N, and Van de Ven WJM. Genomics 36: 118-129, 1996). Using a specific anti-LPP antibody, we showed, in Western blots and with immunofluorescence microscopy, the selective expression of LPP in vascular and visceral smooth muscles (approximately 0.5-1 ng/microg total protein). In other mature (noncultured) tissues, including heart and skeletal muscle, the protein is present only in trace amounts and is closely correlated with the levels of the smooth muscle marker alpha-actin. In freshly isolated guinea pig bladder smooth muscle cells, immunofluorescence images showed LPP as linear arrays of punctate, longitudinally oriented staining superimposed with vinculin staining on the plasma membrane surface. A corresponding pattern of periodic labeling at the membrane in transverse sections of bladder smooth muscle suggested an association of LPP with peripheral dense bodies. In cultured rat aortic smooth muscle cells, LPP colocalized with vinculin at focal adhesions but not with p120 catenin or alpha-actinin. Overexpression of the protein increased EGF-stimulated migration of vascular smooth muscle cells in Transwell assays, suggesting the participation of LPP in cell motility. The Rho-kinase inhibitor Y-27632 dissociated focal adhesions and LPP staining at the cell periphery and enhanced the nuclear accumulation of LPP induced by leptomycin B, indicating that LPP has a potential for relocating to the nucleus through a shuttling mechanism that is sensitive to inhibition of Rho-kinase. PMID- 12760909 TI - Imaging-guided percutaneous radiofrequency ablation of solid renal masses: techniques and outcomes of 38 treatment sessions in 32 consecutive patients. AB - OBJECTIVE: The purpose of this study was to describe the treatment techniques and results of 38 consecutive imaging-guided percutaneous radiofrequency ablations of solid renal masses performed in 32 patients. MATERIALS AND METHODS: Solid renal masses in 32 patients underwent 38 treatment sessions using imaging-guided percutaneous radiofrequency ablation. During 36 sessions, radiofrequency ablation was performed using CT guidance, and two, using sonographic guidance. The average patient age was 76 years (range, 52-87 years), and the average renal mass size was 2.6 cm (range, 1-5 cm). The average number of radiofrequency treatments per solid mass at each session was 2.4 (range, 1-6 treatments), and the average time per treatment was 9.2 min (range, 3-14 min). A single electrode was used in 12 sessions, and a cluster electrode was used in 26 sessions. The average follow-up time was 9 months (range, 1-36 months). RESULTS: Twenty-six of 32 patients had successful treatment of the solid renal mass using percutaneous imaging-guided radiofrequency ablation after one treatment session. Successful treatment was defined as lack of enhancement of the treated region on follow-up CT. Six of 32 patients had residual enhancing tissue after the first treatment session and returned for a second session. Five of these six retreatments were successful. Masses requiring a second treatment session were significantly larger than masses treated in a single session (3.5 vs 2.4 cm, respectively; p = 0.0013). Two patients had perinephric hematomas (which did not require transfusion), and one patient developed a 5-mm skin metastasis at the electrode insertion site, which was resected without recurrence. CONCLUSION: Percutaneous imaging-guided radiofrequency ablation shows promise in the treatment of solid renal malignancies. PMID- 12760911 TI - Bearing false witness. PMID- 12760910 TI - Imaging-guided radiofrequency ablation of solid renal tumors. AB - OBJECTIVE: We performed a retrospective review of imaging-guided radiofrequency ablation of solid renal tumors. MATERIALS AND METHODS: Since May 2000, 35 tumors in 20 patients have been treated with radiofrequency ablation. The size range of treated tumors was 0.9-3.6 cm (mean, 1.7 cm). Reasons for patient referrals were a prior partial or total nephrectomy (nine patients), a comorbidity excluding nephrectomy or partial nephrectomy (10 patients), or a treatment alterative to nephron-sparing surgery (one patient who refused surgery). Tumors were classified as exophytic, intraparenchymal, or central. Sixteen patients had 31 lesions that showed serial growth on CT or MR imaging. Of these 16 patients, four patients with 10 lesions had a history of renal cell carcinoma, and two patients with 11 lesions had a history of von Hippel-Lindau disease. Four patients had incidental solid masses, two of which were biopsied and shown to represent renal cell carcinoma, and the remaining two masses were presumed malignant on the basis of imaging features. Successful ablation was regarded as any lesion showing less than 10 H of contrast enhancement on CT or no qualitative evidence of enhancement after IV gadolinium contrast-enhanced MR imaging. RESULTS: Of the 35 tumors, 22 were exophytic and 13 were intraparenchymal. Twenty-seven of the 35 were treated percutaneously using either sonography (n = 22) or CT (n = 5). Two patients had eight tumors treated intraoperatively using sonography. Patients were followed up with contrast-enhanced CT (n = 18), MR imaging (n = 5), or both (n = 5) with a follow-up range of 1-23 months (mean, 9 months). No residual or recurrent tumor and no major side effects were seen. CONCLUSION: Preliminary results with radiofrequency ablation of exophytic and intraparenchymal renal tumors are promising. Radiofrequency ablation is not associated with significant side effects. Further follow-up is necessary to determine the long-term efficacy of radiofrequency ablation. PMID- 12760912 TI - Preparation of digital images for presentation and publication. PMID- 12760913 TI - Removal of dust and scratches from electronic image files. PMID- 12760914 TI - Percutaneous saline-enhanced radiofrequency ablation of unresectable hepatic tumors: initial experience in 26 patients. AB - OBJECTIVE: The purpose of our study was to evaluate the safety and efficacy of percutaneous saline-enhanced radiofrequency ablation for unresectable primary or metastatic hepatic tumors. SUBJECTS AND METHODS: Twenty-six patients with 15 hepatocellular carcinomas and 33 hepatic metastases (maximum diameter < or = 8.6 cm) were treated; of these, seven tumors in five patients were treated twice. Thus, 44 radiofrequency treatments were performed. Saline-enhanced and impedance controlled radiofrequency ablation (0.5-1.1 mL/min of saline, 15-mm conductive portion of the electrode tip, 25-60 W, 5-43 min) was performed using MR imaging guidance. Coagulation necrosis, volume indexes, morbidity, and complications were assessed. RESULTS: The volume of coagulation necrosis 1-7 days after radiofrequency ablation was 1.6-126.6 cm(3) (median, 18.9 cm(3)), corresponding to coagulation diameters of 1.5-6.2 cm (median, 3.2 cm). The coagulation volume was significantly larger if there were more than four radiofrequency applications (p = 0.006). Tumors of 3 cm or less in diameter were eight times as likely to be successfully completely ablated (p = 0.01) and volume indexes of lesions treated with the patient under general anesthesia were significantly larger than those treated with the patient under conscious sedation (p < 0.001). Major complications occurred in four patients (15%). Incomplete ablation in 19 (35%) of 54 radiofrequency lesions was due to cooling by a large vessel nearby (n = 2) or to low power applied in painful (n = 11) or critical (n = 6) locations. Residual tumor was observed in 14 (58%) of 24 tumors evaluated 6-8 months after radiofrequency ablation. CONCLUSION: Percutaneous saline-enhanced and impedance controlled radiofrequency ablation can be effective in the treatment of unresectable hepatic tumors and minimizes potential carbonization. A greater number of radiofrequency applications, general anesthesia, and increasing experience provide significantly better results. PMID- 12760916 TI - Percutaneous sonographically guided microwave coagulation therapy for hepatocellular carcinoma: results in 234 patients. AB - OBJECTIVE: The objective of our study was to evaluate the long-term results of percutaneous microwave coagulation therapy for the treatment of hepatocellular carcinoma in a large patient population. SUBJECTS AND METHODS: Survival rates were determined in 234 patients with 339 nodules of hepatocellular carcinoma who had undergone percutaneous microwave coagulation therapy (208 men, 26 women; mean age, 54.8 years; mean tumor size, 4.1 +/- 1.9 cm; range, 1.2-8.0 cm; mean follow up period, 27.9 months). Patients were those who had been rejected as candidates for surgery by the surgery department, who fit our study's criteria, and who agreed to participate. After baseline imaging studies were performed, the patients were followed up using the same combination of imaging (sonography, CT, or MR imaging) and posttreatment biopsy. RESULTS: After percutaneous microwave coagulation therapy, color Doppler flow signals disappeared in 92.0% (263/286) of the lesions. No enhancement was apparent in 89.2% (190/213) and 89.1% (41/46) of the lesions on contrast-enhanced CT and MR imaging, respectively. Posttreatment biopsies of 194 nodules showed no evidence of surviving tumor tissue in 180 nodules (92.8%). Resections of six lesions revealed complete tumor necrosis in five. The 1-, 2-, 3-, 4-, and 5-year cumulative survival rates were 92.70%, 81.60%, 72.85%, 66.37%, and 56.70%, respectively. The relationships between survival curves and the degree of hepatocellular carcinoma tumor differentiation and between survival curves and tumor size were statistically significant (p = 0.021). No severe complications were seen. CONCLUSION: Sonographically guided microwave coagulation proved to be safe and effective for the treatment of hepatocellular carcinoma. This therapy resulted in a high percentage of cases without evidence of residual tumor and satisfactory long-term results. PMID- 12760917 TI - Importance of being sure of diagnosis! PMID- 12760918 TI - Do surgical clips interfere with radiofrequency thermal ablation? AB - OBJECTIVE: This study sought to evaluate whether surgical clips affect tissue conductivity and thereby alter the induction of radiofrequency ablation lesions and to determine whether therapy is safe after previous placement of clips in the liver. MATERIALS AND METHODS: An ex vivo porcine hepatic model was used. Three clips were placed around a radiofrequency electrode at 10, 20, and 30 mm from the point of insertion. Clips were arranged in a plane either perpendicular or parallel to the electrode track. After placement of the liver specimen on a grounding pad, radiofrequency energy was applied in a standardized manner for 5 min. Lesion growth and morphology were documented for each minute. RESULTS: Radiofrequency lesions appeared circular and homogeneous after 5 min. Lesion diameter perpendicular to the radiofrequency electrode averaged 30 mm. However, lesion formation was irregular during the early phase of the radiofrequency ablation. The lesion extended irregularly toward the 1-cm clip after 60 sec of ablation. During the second minute, a distinct lesion was observed around the clip 1 cm from the electrode; the primary lesion had not yet reached the clip. During the final 3 min, the primary lesion reached the 1-cm clip and ultimately incorporated the satellite lesion. No lesions were detected surrounding the more distant clips. CONCLUSION: Our data suggest that with the parameters applied in our study, radiofrequency ablation can be safely performed in patients with implanted clips. No aberrant conduction is observed around surgical clips that are located 20 mm and further from the radiofrequency electrode. PMID- 12760919 TI - Diaphragmatic perforation and hernia after hepatic radiofrequency ablation. PMID- 12760920 TI - Usefulness of sonographic guidance during percutaneous biopsy of mesenteric masses. AB - OBJECTIVE: The objective of our study was to determine the usefulness of sonographic guidance for biopsy of mesenteric masses. MATERIALS AND METHODS: Twenty-five sonographically guided percutaneous biopsies of mesenteric masses were performed in 23 patients. Biopsies were performed with an 18-, 20-, or 22 gauge self-aspirating needle or core biopsy device. Final pathology results and patient medical records were reviewed for biopsy accuracy and complications. A biopsy was considered successful if a specific benign or malignant diagnosis was rendered by the pathologist or if surgical-pathologic confirmation was obtained. RESULTS: Open surgical biopsy was performed after sonographically guided biopsy in 13 patients and led to 12 concordant diagnoses (nine true-positives and three true-negatives) and one discordant diagnosis (false-negative). Specific pathologic diagnosis was rendered for the 10 percutaneous biopsies that were not confirmed by surgical biopsy: five biopsies matched known primary malignancies, consistent with metastases; four biopsies revealed primary tumors, and one biopsy revealed chronic inflammation (nine true-positives and one true-negative). Two biopsies were nondiagnostic because of insufficient material (n = 1) and necrotic tumor (n = 1). In the biopsies with diagnostic tissue specimens, sonographically guided biopsy achieved a sensitivity of 95% (18/19) and specificity of 100% (4/4) for allowing neoplastic tissue to be distinguished from nonneoplastic tissue. Complications included a mesenteric hematoma and abdominal wall cellulitis. CONCLUSION: Percutaneous biopsy of mesenteric masses is a useful and safe procedure. PMID- 12760921 TI - MR imaging-guided adrenal biopsy using an open low-field-strength scanner and MR fluoroscopy. AB - OBJECTIVE: The aim of our study was to test the feasibility and specific properties of MR imaging-guided adrenal biopsy using an open 0.2-T scanner and MR fluoroscopic fast imaging with steady-state free precession sequences. CONCLUSION: MR imaging-guided biopsy of the adrenal gland is feasible and safe. In all patients, appropriate specimens were obtained with full diagnostic yield and accuracy. MR fluoroscopy is particularly useful to establish an oblique paravertebral access without pleural transgression. For final needle placement, supplementary breath-hold multislice sequences are required in most cases. PMID- 12760922 TI - Abdominal myomectomy versus uterine fibroid embolization in the treatment of symptomatic uterine leiomyomas. AB - OBJECTIVE: The purpose of this study was to compare treatment efficacy and complications of abdominal myomectomy with those of uterine fibroid embolization in women with symptomatic uterine fibroids. MATERIALS AND METHODS: We analyzed the outcomes of 111 consecutive patients who underwent abdominal myomectomy (n = 44) or fibroid embolization (n = 67) over a 30-month period. The mean ages of the two groups were 37.7 years (range, 28-48 years) and 44.2 years (range, 31-56 years), respectively. A questionnaire and review of medical records assessed all procedure-related complications and changes in symptoms. Length of hospital stay, time until resumption of daily activities, and pain medication requirements after the procedure were also analyzed. RESULTS: Follow-up times for the myomectomy and embolization groups were 14.6 and 14.3 months, respectively. The respective observed success rates in abdominal myomectomy and uterine fibroid embolization patients were 64% versus 92% for menorrhagia (p < 0.05), 54% versus 74% for pain (not significant), and 91% versus 76% for mass effect (p < 0.05). The complication rates were 25% (abdominal myomectomy) and 11% (uterine fibroid embolization) (p < 0.05). The respective secondary end points for the two procedures were 2.9 versus 0 days mean hospital stay, 8.7 versus 5.1 days of narcotics use, and 36 versus 8 days until resumption of normal activities. These differences were all statistically significant. CONCLUSION: Uterine fibroid embolization is a less invasive and safer treatment option in women with symptomatic leiomyomas than myomectomy. Menorrhagia may be better controlled with embolization, and myomectomy may be a better option in patients with mass effect. Both procedures were equally effective in controlling pain. PMID- 12760923 TI - Love is in the air. PMID- 12760924 TI - Added benefit of thoracic aortography after transarterial embolization in patients with hemoptysis. AB - OBJECTIVE: The purpose of this study was to examine the usefulness of thoracic aortography performed after transarterial embolization in identifying additional arteries responsible for causing hemoptysis. SUBJECTS AND METHODS: Between March 2000 and November 2001, we prospectively performed thoracic aortography after transarterial embolization in 76 patients with hemoptysis. Underlying diseases included tuberculosis (n = 34), bronchiectasis (n = 30), emphysema (n = 4), bronchitis (n = 4), aspergillosis (n = 3), and lung cancer (n = 1). Initially, angiography of bronchial and other systemic arteries possibly contributing to hemoptysis was performed with embolization. After completion of the embolization, thoracic aortography was performed, with the tip of the catheter located just distal to the origin of the left subclavian artery. RESULTS: A total of 200 arteries (52 right bronchial, 40 left bronchial, six common bronchial, 76 intercostal, 11 inferior phrenic, six thoracodorsal, eight internal mammary, and one thyrocervical) were identified either at the initial embolization or on thoracic aortography as being responsible for causing hemoptysis. Among them, 29 arteries (14.5%) that were not included on the initial selection for embolization were later identified on postembolization thoracic aortography. There were two right bronchial, three left bronchial, eight inferior phrenic, and 16 intercostal arteries. CONCLUSION: The inferior phrenic and intercostal arteries were often missed on routine transarterial embolization in patients with hemoptysis. Postembolization thoracic aortography is useful for monitoring the effectiveness of embolization and for improving the detection of arteries contributing to hemoptysis. PMID- 12760925 TI - Imaging and intervention in the hepatic veins. PMID- 12760926 TI - Preoperative evaluation of hepatocellular carcinoma: combined use of CT with arterial portography and hepatic arteriography. AB - OBJECTIVE: This study was undertaken to determine the usefulness of combined CT during arterial portography and CT hepatic arteriography in the preoperative evaluation of patients with known or suspected hepatocellular carcinoma and to describe the findings on CT during arterial portography and CT hepatic arteriography by which hepatocellular carcinomas may be differentiated from pseudolesions. SUBJECTS AND METHODS: This study included 137 patients who underwent combined CT during arterial portography and CT hepatic arteriography for the preoperative evaluation of known or suspected hepatocellular carcinoma. The images were prospectively evaluated to identify focal hepatic lesions and their differential diagnoses (hepatocellular carcinoma versus pseudolesion). We assessed the diagnostic accuracy of our prospective interpretation by comparing the interpretations with the results of histopathology or follow-up imaging. We also retrospectively analyzed imaging features seen on CT during arterial portography and CT hepatic arteriography-the size, shape, and location of the lesion within the liver; attenuation of the lesion; and opacification of the peripheral portal vein branches on CT hepatic arteriography. RESULTS: One hundred and forty-nine hepatocellular carcinomas (75 lesions confirmed at histopathology and 74 lesions on follow-up imaging) were found in 120 patients, and 104 pseudolesions (15 lesions confirmed at histopathology and 89 lesions on follow-up imaging) were found in 91 patients. The sensitivity of our prospective interpretations was 98.7%, and the specificity of our prospective interpretations was 90.4%. Our positive and negative predictive values were 93.6% and 97.9%, respectively. We found that hepatocellular carcinomas were larger, more frequently nodular, and more likely to be located intraparenchymally than were the pseudolesions (p < 0.01). Opacification of the peripheral portal vein branches on CT hepatic arteriography was detected in 36 pseudolesions (34.6%) but in none of the hepatocellular carcinomas (p < 0.01). CONCLUSION: Combining CT during arterial portography and CT hepatic arteriography is useful for the preoperative evaluation of patients with known or suspected hepatocellular carcinoma. Familiarity with the imaging features of hepatocellular carcinomas and pseudolesions can help in the accurate differentiation of hepatocellular carcinomas from pseudolesions. PMID- 12760927 TI - Hepatic subcapsular steatosis in response to intraperitoneal insulin delivery: CT findings and prevalence. AB - OBJECTIVE: The aim of this study was to investigate the CT findings and prevalence of hepatic subcapsular steatosis in patients undergoing peritoneal dialysis with intraperitoneal insulin delivery. CONCLUSION: Hepatic subcapsular steatosis appeared as subcapsular nodules and often rindlike areas of low attenuation in seven (18%) of 39 patients who received intraperitoneal insulin with their peritoneal dialysate. Cessation of intraperitoneal insulin therapy led to reversal of the steatosis in three patients. PMID- 12760928 TI - Fitz-Hugh-Curtis syndrome: multidetector CT findings of transient hepatic attenuation difference and gallbladder wall thickening. PMID- 12760929 TI - Anorectal gastrointestinal stromal tumors: CT and MR imaging features with clinical and pathologic correlation. AB - OBJECTIVE: The purpose of this study was to describe the imaging features of anorectal gastrointestinal stromal tumors with clinical and pathologic correlation. CONCLUSION: Anorectal gastrointestinal stromal tumors are mesenchymal neoplasms that typically arise in the muscularis propria of the intestinal wall. The cross-sectional imaging appearance is that of a well-defined mural mass that may have an exophytic component and may invade adjacent structures. A prominent intraluminal component is a rare feature. PMID- 12760930 TI - Videofluoroscopic studies of swallowing dysfunction and the relative risk of pneumonia. AB - OBJECTIVE: The purpose of our investigation was to determine the relationship between the degree of swallowing dysfunction observed on barium studies and the likelihood of developing pneumonia in a large series of patients. MATERIALS AND METHODS: The findings on videofluoroscopic swallowing studies in 381 patients were used to classify these patients into one of five groups: those with normal swallowing; those with abnormal swallowing but no laryngeal penetration or tracheobronchial aspiration; those with laryngeal penetration; those with tracheobronchial aspiration; and those with silent tracheobronchial aspiration. Clinical data were also reviewed to determine how many patients had developed pneumonia during the 6 months before or after the barium studies. The data were then analyzed to determine whether the risk of developing pneumonia increased significantly with each level of swallowing dysfunction seen on barium studies. RESULTS: No significant difference was found in the frequency of pneumonia in patients with abnormal swallowing but no laryngeal penetration or tracheobronchial aspiration compared with patients with normal swallowing on barium studies (p = 0.85). In contrast, patients with laryngeal penetration, tracheobronchial aspiration, or silent tracheobronchial aspiration were approximately four times (p = 0.008), 10 times (p < 0.0001), and 13 times (p < 0.0001), respectively, more likely to develop pneumonia than those with normal swallowing. CONCLUSION: Our findings indicate that the likelihood of developing pneumonia is directly related to the degree of swallowing dysfunction seen on videofluoroscopic studies. Patients with no laryngeal penetration-regardless of whether they had normal or abnormal swallowing-have the lowest risk of developing pneumonia. Patients with laryngeal penetration, tracheobronchial aspiration, or silent tracheobronchial aspiration are, in increasing order of magnitude, significantly more likely to develop pneumonia than patients with normal swallowing. PMID- 12760931 TI - Regression of abdominal visceral aneurysms in polyarteritis nodosa: CT findings. PMID- 12760933 TI - Diagnosis, staging, and surveillance of cervical carcinoma. PMID- 12760934 TI - Multidetector CT angiography for preoperative evaluation of living laparoscopic kidney donors. AB - OBJECTIVE: The purpose of this study was to determine the accuracy of multidetector CT (MDCT) angiography as the primary imaging technique in the evaluation of living kidney donors. SUBJECTS AND METHODS: Seventy-four consecutive living kidney donors (30 men, 44 women; mean age, 41.7 years) who underwent MDCT were evaluated. CT examination was performed with 120 mL of IV contrast material at an injection rate of 3 mL/sec and a pitch of 6. In every case, arterial and venous phase volumetric data sets were acquired at 25 and 55 sec, respectively. Scans were reconstructed at 1-mm intervals for three dimensional (3D) imaging using a volume-rendering technique. Axial CT images and 3D CT angiography were evaluated prospectively by one reviewer and retrospectively by two reviewers who had no knowledge of surgical results. Surgical correlation for the location of primary and accessory renal arteries, early branching of the renal arteries, and renal vein anomalies was made. RESULTS: Seventy-two subjects underwent left nephrectomy, and two subjects underwent right nephrectomy because supernumerary left renal arteries were detected on preoperative CT angiography. Eighteen supernumerary renal arteries (two arteries to 16 kidneys and three arteries to one kidney) to 74 kidneys underwent nephrectomy. CT and surgical findings agreed in 93% of subjects (the average of three reviewers; range, 89-97%). Two small accessory renal arteries were missed by all three reviewers. Those arteries were diminutive and were thought to be insignificant by the surgeons. Early branching of the renal arteries was shown in 14 arteries, and CT and surgical findings agreed in 96% (the average of three reviewers; range, 93-97%). Renal vein anomalies were present in eight subjects, and CT and surgical findings agreed in 99% of the cases (range, 96-100%). CONCLUSION: MDCT angiography is highly accurate for detecting vascular anomalies and providing anatomic information for laparoscopic living donor nephrectomy. PMID- 12760935 TI - Diagnostic accuracy of phase-inversion tissue harmonic imaging versus fundamental B-mode sonography in the evaluation of focal lesions of the kidney. AB - OBJECTIVE: We compared phase-inversion tissue harmonic imaging with fundamental B mode sonography in the evaluation of focal lesions of the kidney. SUBJECTS AND METHODS: For our prospective study, 114 patients underwent sonography of the kidneys in both modes, fundamental B-mode sonography and phase-inversion tissue harmonic imaging, in a randomly chosen scanning order. Imaging parameters were standardized. Sonographic diagnoses were made under real-time conditions by the examining radiologist. All sonographic diagnoses were compared with a diagnostic reference modality: contrast-enhanced CT, contrast-enhanced MR imaging, or histopathology. Three radiologists different from the examiners evaluated overall image quality, lesion conspicuity, and fluid-solid differentiation for both modalities using hard-copy images. RESULTS: In 70 patients, fundamental B-mode sonography as the first technique depicted 73 of 111 lesions 10 mm or larger and enabled 71 lesions to be correctly characterized (sensitivity, 65.8%; accuracy, 64.0%). As the first mode, phase-inversion tissue harmonic imaging depicted 57 of 65 focal lesions and enabled 54 lesions to be accurately classified in 44 patients (sensitivity, 87.7%; accuracy, 83.1%). The differences in sensitivity and accuracy were statistically significant (95% confidence interval). For overall image quality, lesion conspicuity, and fluid-solid differentiation phase inversion harmonic imaging was superior to fundamental B-mode sonography (p < 0.0001). CONCLUSION: Phase-inversion tissue harmonic imaging is superior to fundamental B-mode sonography in the sonography of focal kidney lesions because phase-inversion tissue harmonic imaging has better overall image quality, lesion conspicuity, and fluid-solid differentiation. In six cases, phase-inversion tissue harmonic imaging added crucial diagnostic information that changed patient management. PMID- 12760936 TI - Characterization of adrenal tumors by chemical shift fast low-angle shot MR imaging: comparison of four methods of quantitative evaluation. AB - OBJECTIVE: The aim of our study was to assess quantitative methods of distinguishing adenomas from malignant adrenal lesions using chemical shift fast low-angle shot MR imaging. MATERIALS AND METHODS: We assessed 102 adrenal tumors in 88 patients (27 hyperfunctioning and 44 nonhyperfunctioning adenomas, 19 metastases, nine pheochromocytomas, and three other adrenal tumors) using chemical shift MR imaging. On the chemical shift imaging, signal intensity index, calculated as [(signal intensity on in-phase imaging - signal intensity on opposed-phase imaging) / (signal intensity on in-phase imaging)] x 100%, was compared with the adrenal-to-spleen ratio, adrenal-to-muscle ratio, and adrenal to-liver ratio for signal change on opposed-phase fast low-angle shot MR imaging. The tissues in the spleen, paraspinal muscle, and liver were reference tissues. RESULTS: The signal intensity index had several advantages over the other three parameters calculated. We found no overlap in indexes between adenomas and metastatic tumors. The accuracy in distinguishing adenomas from metastatic tumors was 100% if the cutoff value of the signal intensity index selected was 11.2 16.5%. CONCLUSION: The signal intensity index is the most reliable evaluation method for differentiating adrenal adenomas from metastatic adrenal tumors. PMID- 12760937 TI - Multidetector CT appearance of a novel contraceptive device: the vaginal ring. PMID- 12760938 TI - Six cases of acute central pulmonary embolism revealed on unenhanced multidetector CT of the chest. AB - OBJECTIVE: Our purpose was to describe the imaging findings of central pulmonary embolism on unenhanced multidetector CT (MDCT) of the chest. CONCLUSION: Unenhanced MDCT of the chest is often performed for the evaluation of nonspecific chest symptoms. Awareness of the rare finding of a high-attenuation centrally located pulmonary embolism on unenhanced MDCT is important because acute pulmonary embolism may be identified when it is not suspected clinically, and such detection can determine further imaging needs and allow the timely initiation of appropriate therapy. PMID- 12760939 TI - CT-guided transthoracic needle aspiration biopsy of small (< or = 20 mm) solitary pulmonary nodules. AB - OBJECTIVE: The purpose of our study was to determine the diagnostic accuracy and to analyze the factors influencing the diagnostic accuracy and incidences of pneumothorax and chest tube insertion rates for percutaneous CT-guided needle biopsy of small (< or = 20 mm) solitary pulmonary nodules. SUBJECTS AND METHODS: One hundred sixty-two patients with 162 small solitary pulmonary nodules underwent CT-guided transthoracic needle aspiration biopsy. The overall diagnostic accuracy, pneumothorax rate, and chest tube insertion rate were calculated. Factors influencing the diagnostic accuracy and pneumothorax rate were statistically evaluated. Influencing factors, diagnostic accuracies, pneumothorax rates, and chest tube insertion rates were statistically compared. RESULTS: Overall diagnostic accuracy, pneumothorax rate, and chest tube insertion rate were 77.2%, 28.4%, and 2.5%, respectively. Diagnostic accuracy was significantly affected by length of needle path and lesion size (p < 0.05). The pneumothorax rate was significantly affected by the percentage of predicted forced expiratory volume in 1 sec, the number of punctures, and the needle path length (p < 0.05). The chest tube insertion rate was significantly affected by the number of punctures (p < 0.05). For diagnostic accuracy, needle path lengths of 40 mm or less and lesion sizes greater than 10 mm were significantly more accurate than other factors (p < 0.05). For pneumothorax rates, a percentage of predicted forced expiratory volume in 1 sec of greater than 70%, a single puncture, and a needle path length of 40 mm or less were significantly lower than other factors (p < 0.05). CONCLUSION: CT-guided transthoracic needle aspiration biopsy is a useful diagnostic tool for small solitary pulmonary nodules smaller than 20 mm in diameter. The diagnostic accuracy is significantly improved for large (> 10 mm) lesion size and short (< or = 40 mm) needle path length. PMID- 12760940 TI - CT of blunt tracheal rupture. PMID- 12760941 TI - Metastatic angiosarcoma of the lung: spectrum of CT findings. AB - OBJECTIVE: The purpose of our study was to summarize the CT features of pulmonary metastases in angiosarcoma in 24 patients. CONCLUSION: A variety of CT features were associated with metastatic angiosarcoma of the lung. The common CT manifestations of metastatic angiosarcoma were multiple solid nodular lesions and multiple thin-walled cysts that were often accompanied by hemorrhagic change. PMID- 12760942 TI - Mammography and subsequent whole-breast sonography of nonpalpable breast cancers: the importance of radiologic breast density. AB - OBJECTIVE: Our purpose was to determine the contribution of mammography followed by sonography for the detection of nonpalpable breast cancers in Breast Imaging Reporting and Data System (BI-RADS) density grades 1-4 breasts, in grades 1 and 2 breasts, and in grades 3 and 4 breasts. MATERIALS AND METHODS: The results of physical, mammographic, and sonographic examinations performed in 4236 patients were reviewed to determine the sensitivities of mammography and sonography for the detection of nonpalpable breast cancers and to calculate the relative risk for detecting nonpalpable breast cancers using sonography in comparison with mammography in density grades 1-4, grades 1 and 2, and grades 3 and 4 breasts. Sonography was performed after mammographic interpretation. RESULTS: Sensitivities of mammography and subsequent sonography for the detection of nonpalpable breast cancers were 69% and 88% in grades 1-4, 80% and 88% in grades 1 and 2, and 56% and 88% in grades 3 and 4 breasts, respectively. The relative risk for detecting nonpalpable breast cancers using sonography was statistically significantly greater than that for detecting nonpalpable breast cancers using mammography in grades 1-4 (relative risk, 1.29; p = 0.024) and in grades 3 and 4 (relative risk, 1.57; p = 0.013) but not in grades 1 and 2 (relative risk, 1.1; p = 0.445) breasts. CONCLUSION: Sonography is a useful adjunct after mammography for the detection of nonpalpable breast cancer, particularly in the dense breast. PMID- 12760943 TI - Solid variant of aneurysmal bone cysts in long tubular bones: giant cell reparative granuloma. AB - OBJECTIVE: The purpose of this study was to determine the age distribution, location, and imaging features of histologically proven solid variants of aneurysmal bone cysts in long tubular bones. MATERIALS AND METHODS: We performed a retrospective review of imaging studies of histologically proven solid aneurysmal bone cysts in long bones between 1961 and 2001. There were 30 cases comprising 29 radiographic, six CT, and eight MR imaging examinations. The lesions were evaluated for bone involved, location within a long bone, matrix, size, soft-tissue mass, and MR imaging characteristics. The imaging findings were correlated with the histologic findings. RESULTS: The patients were 17 females and 13 males ranging in age from 2 to 58 years (mean, 18 years). The bones involved were the femur (n = 10), the ulna (n = 7), the tibia (n = 7), the humerus (n = 2), the radius (n = 2), and the fibula (n = 2). The lesions were five juxtaarticular, 13 metaphyseal, one diametaphyseal, and 11 diaphyseal. The location was eccentric in 20 cases, of which two were intracortical and two periosteal, and central in 10. Lesion size varied between 1 and 7 cm. Thirty three percent of lesions were nonaneurysmal. Four lesions were mineralized. A soft-tissue mass was present in four cases. Four lesions showed a permeative lytic pattern simulating a malignant process. Unusual findings included periosteal reaction and development of a solid aneurysmal bone cyst in a preexisting fracture. MR imaging showed solid elements in all cases and pronounced edema in 50% of cases. CONCLUSION: Solid aneurysmal bone cyst is a reactive nonneoplastic bone lesion with varied imaging characteristics; one third of lesions are nonaneurysmal. PMID- 12760945 TI - Differential diagnosis of benign peripheral lipoma from well-differentiated liposarcoma on MR imaging: is comparison of margins and internal characteristics useful? AB - OBJECTIVE: Our objective was to evaluate the reliability of MR imaging in distinguishing between benign lipoma and well-differentiated liposarcoma. MATERIALS AND METHODS: The MR images of 35 pathologically proven benign lipomas in 35 patients and 23 well-differentiated liposarcomas in 17 patients were retrospectively reviewed. T1-, T2-, and fat-suppressed T1-weighted images were obtained after administration of gadopentetate dimeglumine. Margins and internal characteristics revealed on the MR images and the degree of contrast enhancement of septa were evaluated. These MR imaging findings were compared for well differentiated liposarcomas and benign lipomas. RESULTS: Completely irregular margins were recognized only in benign lipomas with a pathologic diagnosis of infiltrating lipoma. All tumors without a recognizable nonadipose component were benign lipomas (p < 0.05). As for the well-differentiated liposarcomas, thick septa and nodular or patchy nonadipose components were present more frequently in deep and retroperitoneal lesions than in subcutaneous lesions (p < 0.01). No cases showed only thin septa in the deep lesions of well-differentiated liposarcoma, and all cases showed thick septa or nodular or patchy nonadipose components. The septa in well-differentiated liposarcomas enhanced more strongly than did those in benign lipomas. The septa showed no enhancement relative to muscle in 11 of 19 benign lipomas, whereas the septa showed moderate or marked enhancement in all well-differentiated liposarcomas (p < 0.01). CONCLUSION: Careful assessment of margins and internal characteristics on MR imaging can be a useful aid in further distinguishing between biologically different benign lipoma and well-differentiated liposarcoma. PMID- 12760947 TI - Radiography of the PROSTALAC (prosthesis with antibiotic-loaded acrylic cement) orthopedic implant. PMID- 12760946 TI - Osseous change adjacent to soft-tissue hemangiomas of the extremities: correlation with lesion size and proximity to bone. AB - OBJECTIVE: The purpose of this study was to describe the findings of MR imaging and radiographic changes that occur in osseous structures adjacent to soft-tissue hemangiomas of the extremities and to correlate them with patient symptomatology, the size of the hemangiomas, and their proximity to adjacent bone. MATERIALS AND METHODS: We retrospectively reviewed the radiographs and MR images of 35 patients with soft-tissue hemangiomas of the extremities. The pattern and extent of the osseous change were categorized as periosteal, cortical, or medullary. Symptomatology, size, and proximity of the hemangioma to the adjacent bone were compared with the presence or absence of osseous change. Statistical analysis was performed using the Student's t test. RESULTS: Osseous change was noted on radiographs in 13 (37%) of 35 patients and on MR images in 11 (31%) of 35 patients with a total of 14 patients (40%) showing osseous change on at least one study. Periosteal change was present in eight (23%) of 35 patients; cortical change, in 11 (31%) of 35 patients; and medullary change, in 10 (29%) of 35 patients. Direct contact between the soft-tissue hemangioma and the adjacent bone was seen in 13 of 14 patients with osseous change. In those patients without osseous change, the average distance between the soft-tissue hemangioma and bone was 1.06 cm (range, 0-4 cm). No correlation was found between symptoms and the presence of osseous change. CONCLUSION: Soft-tissue hemangiomas of the extremities frequently result in adjacent osseous change that can be categorized as either periosteal, cortical, or medullary. Only medullary changes correspond with hemangioma size, whereas all three categories of change correlate with the proximity of the hemangioma to the adjacent bone. The presence of osseous change does not correlate with patient symptomatology. PMID- 12760948 TI - Four-channel multidetector CT in facial fractures: do we need 2 x 0.5 mm collimation? AB - OBJECTIVE: Our aim was to optimize acquisition protocols and multiplanar reformation algorithms for the evaluation of facial fractures using multidetector CT (MDCT) and to determine whether 2 x 0.5 mm collimation is necessary. MATERIALS AND METHODS: A cadaveric head with artificial blunt facial trauma was examined using a four-channel MDCT scanner. The influence of acquisition parameters (collimation, 2 x 0.5 mm, 4 x 1 mm, 4 x 2.5 mm; tube current, 120 mAs, 90 mAs, 60 mAs), image reconstruction algorithms (standard vs ultra-high-resolution modes; reconstructed slice thicknesses, 0.5 mm, 1 mm, 3 mm; increment, 0.3 mm, 0.6 mm, 1.5 mm), and reformation algorithms (slice thicknesses, 0.5 mm, 1 mm, 3 mm; overlap, 0.5 mm, 1 mm, 3 mm) on detectability of facial fractures in multiplanar reformations with MDCT was analyzed. RESULTS: Fracture detection was significantly higher with thin multiplanar reformations (0.5 and 0.5 mm, 1 and 0.5 mm, and 1 and 1 mm) (p < or = 0.014) acquired with 2 x 0.5 mm collimation (p < or = 0.046) in ultra-high-resolution mode (p < 0.0005) with 120 mAs (p < or = 0.025). Interobserver variability showed very good agreement (kappa > or = 0.942). Non-ultra-high-resolution mode, lower milliampere-seconds, and thick multiplanar reformations (3 and 0.5 mm, 3 and 1 mm, and 3 and 0.5 mm) showed significantly decreased fracture detectability. CONCLUSION: Although thin multiplanar reformations obtained from thin collimation (2 x 0.5 mm) are statistically superior for the detection of subtle fractures, 4 x 1 mm collimation is sufficient for routine diagnostic evaluation. Ultra-high resolution mode with 120 mAs is mandatory for detection of clinically relevant fractures. PMID- 12760949 TI - MR imaging of diverse manifestations of nasopharyngeal carcinomas. PMID- 12760950 TI - Efficacy of sonographically guided percutaneous ethanol injection for treatment of thyroid cysts versus solid thyroid nodules. AB - OBJECTIVE: Sonographically guided percutaneous ethanol injection has been recently proposed as a treatment for nonfunctioning benign thyroid nodules such as cysts or solid nodules. The objective of this study was to compare the efficacy of ethanol injection in thyroid cysts and solid nodules. SUBJECTS AND METHODS: We studied 20 patients with simple or complex thyroid cysts and 22 patients with solid thyroid nodules. All lesions were confirmed by fine-needle aspiration biopsy to be benign. The mean volume of the instilled absolute ethanol (99.9%) was 62.2% of the whole tumor volume for solid nodules and 63.4% of the cystic volume for cysts. Follow-up sonography was performed 1-6 months (mean, 4.4 months for cysts and 4.6 months for solid nodules) after the procedure. RESULTS: The mean volume reduction rate for cysts (65%) was greater than that for solid nodules (38.3%) (p < 0.01, Student's t test). The volume of the instilled ethanol correlated significantly with the volume reduction rate of cysts but not with that of solid nodules (p < 0.01, Student's t test). CONCLUSION: Sonographically guided percutaneous ethanol injection is more effective for thyroid cysts than for solid thyroid nodules. PMID- 12760951 TI - Evaluation of emergency CT scans of the head: is there a community standard? AB - OBJECTIVE: This study was designed to assess the accuracy of general radiologists in the interpretation via teleradiology of emergency CT scans of the head. MATERIALS AND METHODS: We studied the interpretations of 716 consecutive emergency CT scans of the head by a group of 15 board-certified general radiologists practicing in the community (as opposed to an academic setting). The scans were sent via teleradiology, and the preliminary interpretations were made. Three of the general radiologists were functioning as nighthawks, and the remaining 12 were acting as on-call radiologists in addition to their normal daytime duties. Each CT examination was interpreted by one of five neuroradiologists the day after the initial interpretation had been performed. The findings of the final interpretation and the preliminary interpretation were categorized as showing agreement, insignificant disagreement, or significant disagreement. The reports in the two categories indicating disagreement were reviewed and reclassified by a consensus of three university-based neuroradiologists. RESULTS: Agreement between the initial interpretation by the general radiologist and the final interpretation by the neuroradiologist was found in 95% of the CT scans. The interpretations were judged to show insignificant disagreement in 3% (23/716) of the scans and to show significant disagreement in 2% (16/716). Of the 16 significant errors, five were false positive findings and 11 were false-negative findings. Forty-seven CT scans depicted significant or active disease, and in 11 (23%) of these scans, the final report differed significantly from the preliminary interpretation. Three patients had pituitary masses, none of which had been described on the preliminary interpretation. CONCLUSION: The rate of significant discordance between board certified on-call general radiologists and neuroradiologists in the interpretation of emergency CT scans was 2%, which was comparable to previously published reports of residents' performance. The pituitary gland may be a blind spot, and additional attention should be focused on this area. PMID- 12760952 TI - Correlation of contrast-enhanced power Doppler sonography and conventional angiography of abduction-induced hip ischemia in piglets. AB - OBJECTIVE: The purpose of this study was to determine if enhanced power Doppler sonography can detect early ischemia of the capital femoral epiphysis induced by hip hyperabduction in piglets and to correlate these findings with angiography. MATERIALS AND METHODS: Proximal femoral perfusion was evaluated in 18 studies of 10 piglet hips with unenhanced power Doppler sonography, enhanced power Doppler sonography with IV contrast agent, and digital angiography, in neutral position, hyperabduction, and after release to neutral position. Enhancement ratios between pixel intensities of power Doppler sonography and enhanced power Doppler images in each position were calculated. Angiograms were analyzed for differences in flow with changes in hip position. RESULTS: With the piglet in neutral position, power Doppler sonography revealed few vessels in the femoral head. Contrast administration resulted in a temporary marked increase in the visualization of vessels in the femoral head. Quantitative enhanced power Doppler sonography revealed a marked decrease in pixel intensity with abduction (p < 0.001) that was not apparent on unenhanced studies (p = 0.28). The enhancement ratio decreased from 0.45 (mean +/- SD, +/- 0.26) in neutral position to 0.10 (+/- 0.21) after abduction; it returned to 0.41 (+/- 0.14) after release of abduction (p < 0.001 for each comparison). Angiographic studies in hyperabduction revealed a variable level of ischemia. CONCLUSION: Enhanced power Doppler sonography can be used to visualize the vascular supply to the cartilaginous femoral head in piglets and can detect reversible ischemia induced by hip hyperabduction. These differences correlate with digital angiographic evidence of ischemia. PMID- 12760953 TI - CT screening for lung cancer. PMID- 12760954 TI - SARS and the modern day pony express (the World Wide Web). PMID- 12760956 TI - A unique presentation of cardiac sarcoidosis. PMID- 12760957 TI - Intervertebral disk hydatid cysts: MR imaging findings. PMID- 12760958 TI - Rosai-Dorfman disease presenting as a suspicious breast mass. PMID- 12760959 TI - Pulmonary hematoma resulting from anticoagulant therapy. PMID- 12760960 TI - The airway epithelium as immune modulator: the LARC ascending. PMID- 12760961 TI - Immunostimulatory DNA for asthma: better than eating dirt. PMID- 12760962 TI - Airway epithelial cells release MIP-3alpha/CCL20 in response to cytokines and ambient particulate matter. AB - The initiation and maintenance of airway immune responses in Th2 type allergic diseases such as asthma are dependent on the specific activation of local airway dendritic cells (DCs). The cytokine microenvironment, produced by local cells, influences the recruitment of specific subsets of immature DCs and their subsequent maturation. In the airway, DCs reside in close proximity to airway epithelial cells (AECs). We examined the ability of primary culture human bronchial epithelial cells (HBECs) to synthesize and secrete the recently described CC-chemokine, MIP-3alpha/CCL20. MIP-3alpha/CCL20 is the unique chemokine ligand for CCR6, a receptor with a restricted distribution. MIP 3alpha/CCL20 induces selective migration of DCs because CCR6 is expressed on some immature DCs but not on CD14+ DC precursors or mature DCs. HBECs were stimulated with pro-inflammatory cytokines tumor necrosis factor-alpha and interleukin (IL) 1beta or, because of their critical role in allergic diseases, IL-4 and IL-13. Cells were also exposed to small size-fractions of ambient particulate matter. Each of these stimuli induced MIP-3alpha/CCL20 gene and protein expression. Moreover, these agents upregulated mitogen-activated protein kinase pathways in HBECs. Inhibition of the ERK1/2 pathway or p38 reduced cytokine-induced MIP 3alpha/CCL20 expression. These data suggest a mechanism by which AEC may facilitate recruitment of DC subsets to the airway. PMID- 12760963 TI - Resolution of airway inflammation following ovalbumin inhalation: comparison of ISS DNA and corticosteroids. AB - In this study we have compared the therapeutic effect of the administration of immunostimulatory DNA sequences (ISS) with that of corticosteroids on the resolution of airway inflammation and airway hyperreactivity (AHR) in a mouse model. Mice which had already developed significant levels of eosinophilic airway inflammation 24 h after allergen challenge were then treated with either ISS or corticosteroids, and the effect on AHR and airway inflammation assessed 6 d later. ISS inhibited AHR as effectively as corticosteroids. Combination therapy with ISS and corticosteroids was more effective than monotherapy with either ISS or corticosteroids in inhibiting AHR. In ovalbumin-challenged mice, levels of bronchoalveolar lavage (BAL) eosinophils were significantly reduced with either ISS or corticosteroids. ISS induced significant levels of BAL interferon-gamma, whereas corticosteroids did not induce expression of BAL interferon-gamma. Both ISS and corticosteroids significantly reduced levels of interleukin-5 in BAL, as well as the number of Periodic Acid Schiff-positive airway epithelial cells. Corticosteroids, but not ISS, increased the number of eosinophils in regional mediastinal lymph nodes. Very few apoptotic peribronchial cells were noted following ovalbumin challenge as assessed by TUNEL assay. Corticosteroids, but not ISS, induced an increase in the small number of apoptotic peribronchial cells. The mechanism by which either ISS or corticosteroids inhibit AHR is likely to be mediated by distinct and shared cellular pathways. The combination of the shared and distinct anti-inflammatory pathways may account for the additive effect of ISS and corticosteroids on inhibiting AHR. PMID- 12760964 TI - Interleukin-9 induces mucous cell metaplasia independent of inflammation. AB - Interleukin-9 (IL-9) has been strongly implicated in the pathogenesis of asthma, including the overproduction of mucus, in humans and in animal models. We evaluated the inflammatory changes associated with the upregulation of mucus production by examining the time course of inflammation after daily intratracheal IL-9 administration to naive C57Bl6 mice for 9 d. IL-9 induced an asthmatic phenotype, which in general took several days to develop, as assessed by the measurement of airway hyperresponsiveness, pulmonary inflammation, and serum immunoglobulin E. However, within 24 h of a single dose of IL-9, muc5ac mRNA upregulation occurred, and increased numbers of periodic acid Schiff/Alcian blue positive mucous cells appeared. This response occurred before the development of an inflammatory cell influx and was the result of epithelial metaplasia. It seemed that IL-9 evoked mucous cell metaplasia independent of IL-13 because mRNA tissue evaluation indicated that muc5ac upregulation preceded any increase in IL 13 mRNA expression or detectable levels of IL-13 in the brochoalveolar lavage fluid. Therefore, the upregulation of IL-13 by IL-9 may be responsible for the amplification of mucus production but is not required for its initiation. IL-9 seems to directly stimulate mucous cell metaplasia without the requirement of inflammatory cell influx. PMID- 12760965 TI - The kinetics and pattern of tracheal allograft re-epithelialization. AB - Extensive tracheal defects may pose a life-threatening dilemma. Although tracheal transplantation may represent a reconstructive solution, very little is known regarding the immunobiology and behavior of tracheal allografts. The objective of this study was to assess the pattern and kinetics of re-epithelialization of orthotopic tracheal allografts in immunosuppressed recipients. Thirty-eight age matched mice were randomly assigned to five experimental groups. BALB/c donor tracheal segments were orthotopically transplanted into either syngeneic BALB/c or MHC mismatched allogeneic C57BL/6 recipients with and without immunosuppression. On post-transplant days 7, 14, 28, 48, and 62, animals from each group were evaluated by serial histology, electron microscopy, and serial immunohistochemical analysis for mucosal phenotype, re-epithelialization pattern, and lymphocyte subpopulations. Nonimmunosuppressed recipients underwent recipient derived basal cell re-epithelialization by Day 48, with differentiation into a sparse population of ciliated columnar epithelium by Day 62, whereas immunosuppressed recipients underwent basal cell re-epithelialization 28 d after transplantation and differentiation into a dense population of ciliated columnar epithelium by Day 48. The re-epithelialization process occurred in a definable pattern that was significantly enhanced with the addition of immunosuppression. Orthotopic tracheal transplants undergo progressive re-epithelialization with recipient-derived basal cells that differentiate into ciliated columnar epithelium in a definable pattern that is enhanced with the addition of immunosuppression. PMID- 12760966 TI - Gene expression profiling of the early pulmonary response to hyperoxia in mice. AB - To identify molecular events occurring during the early response to hyperoxia, we measured changes over time in total lung gene expression in C57BL/6 mice during prolonged exposure to > 95% O2. Specifically, differential gene expression of > 8,734 sequence-verified murine complementary DNAs was analyzed after 0, 8, 24, and 48 h of O2 exposure, with additional genes of interest analyzed at 24 h. Of the 385 genes differentially expressed, hyperoxia increased expression of 175 genes (2.0%) and decreased expression of 210 genes (2.3%). The majority of "classic" antioxidant enzymes, including catalase, MnSOD, and Cu-Zn SOD, showed no change in expression during hyperoxia, with a number of other antioxidant enzymes, including glutathione peroxidase, glutathione-S-Transferase (GST) Pi1, GST mu2, and heme oxygenase-1 showing relatively moderate increases. The exception was the heavy metal-binding protein metallothionein, which increased expression over 7-fold after 48 h of O2. We found no change in the expression of a number of known proinflammatory genes after 24 or 48 h of hyperoxia. A large increase in p21 expression was demonstrated, suggesting overall inhibition of cell cycle progression. Increases of the antiapoptotic gene Bcl-XL were counterbalanced by similar increases of the proapoptotic gene BAX. New findings included significant increases in expression of cysteine-rich protein 61(cyr61) at 48 h, suggesting a potential role for this factor in angiogenesis or remodeling of the extra cellular matrix during recovery from hyperoxia. In addition, downregulation of thrombomodulin expression occurred by 24 h and was further decreased at 48 h. Given the importance of thrombomodulin/thrombin interaction in regulating protein C activity, decreases in thrombomodulin may contribute to activation of the coagulation and inflammatory cascades and development of lung injury with hyperoxia. PMID- 12760967 TI - Interaction between respiratory syncytial virus and particulate matter in guinea pig alveolar macrophages. AB - Alveolar macrophages (AM) play a pivotal role in host lung defense mechanisms. Respiratory syncytial virus (RSV) stimulates secretion of proinflammatory cytokines in AM while it suppresses the cell's phagocytic ability. However, exposure of AM to ambient particulate matter (PM10) has been reported to inhibit RSV uptake. The mechanisms involved in the interaction between RSV and PM10 in AM are not known. We hypothesize that the cellular response of AM to RSV and PM10 is dependent on the sequence in which AM are exposed to these agents. In this study, we compared the sequential effect of RSV and PM10 exposure in vitro on the phagocytic function of guinea pig AM, the RSV Yield in AM, and the production of proinflammatory cytokines (interleukin [IL]-6, IL-8, and tumor necrosis factor [TNF]-alpha). The ability of AM to phagocytose PM10 was not affected by sequential exposure to RSV and PM10. RSV Yield was severely decreased in PM10 exposed AM, regardless of sequence of exposure, compared with AM that were not exposed to PM10 (P < 0.004). Exposure of AM to RSV and/or PM10 resulted in enhanced secretion of bioactive TNF-alpha compared with controls (P < 0.02), without synergistic or inhibitory interaction of these agents on TNF-alpha production. By contrast, exposure of AM to PM10 significantly decreased the production of RSV-induced IL-6 (P < 4 x 10(-6)) and IL-8 (P < 0.003). In summary, our findings suggest that PM10 exposure may interfere with mechanisms of RSV replication and viral-induced cytokine production in guinea pig AM, independent of the sequence of exposure to these agents. PMID- 12760968 TI - Priming of eosinophil migration across lung epithelial cell monolayers and upregulation of CD11b/CD18 are elicited by extracellular Ca2+. AB - In patients with asthma, eosinophils are primed and massively infiltrate lung tissues and migrate across epithelia into airways. Using blocking monoclonal antibodies, we found that eosinophil transmigration across a lung epithelial cell monolayer depended on the functions of alphaMbeta2 integrin CD11b/CD18. To study the role of Ca2+ in eosinophil priming and transepithelial migration, we treated eosinophils with eotaxin or thapsigargin (TG), reagents that increase cytoplasmic free Ca2+ concentrations by receptor- or nonreceptor-mediated mechanisms, respectively. Pretreatment of eosinophils with TG enhanced CD11b/CD18-dependent transmigration across lung epithelium. Within minutes, TG time- and dose dependently upregulated the expression of CD11b/CD18 but did not upregulate the expression of alphaL (CD11a) or beta1 (CD29) integrin. The upregulation of CD11b/CD18 expression by eotaxin or TG was prevented when Ca2+ entry was blocked. The priming of eosinophil transmigration by TG was also abrogated by the blockade of Ca2+ entry. Our results indicate that induction of Ca2+ entry by the depletion of Ca2+ from intracellular stores upregulates CD11b/CD18 expression on eosinophils and primes eosinophil transmigration across lung epithelium. Both responses are therefore elicited by extracellular Ca2+. We suggest that, as an important priming signal for human eosinophil functional responses, store operated Ca2+ entry may be one of the underlying mechanisms of eosinophilic inflammation in asthma. PMID- 12760969 TI - Evaluation of inducible costimulator/B7-related protein-1 as a therapeutic target in a murine model of allergic airway inflammation. AB - Given its primary role in the execution of T cell, and especially Th2, effector activity, the inducible costimulator (ICOS)/B7-related protein (RP)-1 costimulatory pathway is currently being heralded as a promising therapeutic target for immune-inflammatory disorders such as asthma. This study investigates the merits of ICOS blockade in a murine model of experimental asthma in which mice are sensitized to ovalbumin (OVA) through the respiratory mucosa. Intraperitoneal treatment of mice with anti-ICOS neutralizing antibody during sensitization resulted in a marked reduction in airway eosinophilia and IL-5 in bronchoalveolar lavage, but had no effect on interleukin (IL)-4, IL-13, and eotaxin content in bronchoalveolar lavage or the production of OVA-specific immunoglobulin E in serum. Cultured splenocytes from mice sensitized to OVA in the context of ICOS ablation produced enhanced levels of IL-4 and IL-5 upon stimulation with OVA, and this correlated with elevated inflammation and immunoglobulin E secretion upon long-term in vivo OVA recall; the deleterious effects ICOS blockade, however, were not associated with reduced IL-10 production by splenocytes. Peculiarly, anti-ICOS intervention during OVA rechallenge had no effect on airway inflammation or immunoglobulin production, despite high levels of ICOS expression on infiltrating CD4+ T cells. This study provides in vivo evidence of an exacerbated long-term immune-inflammatory response following acute ICOS blockade, and suggests that ICOS costimulation is functionally redundant in established allergic disease. PMID- 12760970 TI - C-Jun-NH2-terminal kinase mediates expression of connective tissue growth factor induced by transforming growth factor-beta1 in human lung fibroblasts. AB - Many of the fibrogenic effects of transforming growth factor-beta (TGF-beta) might be mediated by connective tissue growth factor (CTGF). The present study investigates the role of mitogen-activated protein (MAP) kinase in the expression of CTGF mRNA in the human lung fibroblast line, HFL-1. TGF-beta1 enhanced CTGF mRNA levels in a time- and concentration-dependent manner, and this enhancement was also dependent upon transcription. Inhibition of p38 MAP kinase or extracellular signal-regulated kinase (ERK) activation did not affect TGF-beta1 induced CTGF expression. On the other hand, specific inhibitors of phosphatidylinositol 3-kinase (PI3K) suppressed TGF-beta1-induced CTGF expression in a concentration-dependent manner. TGF-beta1 activated c-Jun NH2-terminal kinase (JNK) and p38 MAP kinase, but not ERK in HFL-1 cells. PI3K inhibitors dose dependently suppressed TGF-beta1-induced JNK, but not p38 MAP kinase activation. Finally, JNK1 and JNK2 antisense oligonucleotides attenuated cellular levels of JNK1 and JNK2 protein, respectively, and repressed TGF-beta1-induced CTGF expression. These results suggest that TGF-beta1-induced CTGF mRNA expression is mediated through the JNK-dependent pathway, whereas p38 MAP kinase and ERK pathways minimally contribute. PMID- 12760971 TI - Epidural insertion in anesthetized adults: will your patients thank you? PMID- 12760972 TI - Small risk of serious neurologic complications related to lumbar epidural catheter placement in anesthetized patients. AB - Previous studies have identified pain during needle/catheter placement or during the injection of local anesthetic as a risk factor for the development of persistent paresthesias after regional anesthetic techniques. The performance of regional blockade on anesthetized patients theoretically increases the risk of postoperative neurologic complications, because these patients are unable to respond to painful stimuli. In this study, we evaluated the frequency of neurologic complications in 4298 thoracic surgical patients undergoing lumbar epidural catheter placement while under general anesthesia. Catheters were placed immediately after the induction and tracheal intubation or on completion of the surgical procedure, before emergence. Most epidural catheters (4220, or 98.2%) were used solely for postoperative analgesia; only 78 (1.8%) epidural catheters were used for intraoperative anesthesia. In 4239 (98.6%) patients, an opioid alone was administered. The remaining 56 (1.3%) patients received a local anesthetic or local anesthetic/opioid mixture epidurally. Analgesia was graded as excellent or good in 92.2% of patients. Side effects included sedation in 455 (10.6%), nausea or emesis in 328 (7.6%), pruritus in 116 (2.7%), and respiratory depression (pH or=50 mm Hg) in 308 (7.2%) patients. The mean duration of epidural analgesia was 2.4 +/- 0.8 days (range, 0-10.7 days). There were no neurologic complications, including spinal hematoma, epidural abscess or catheter site infections, radicular symptoms, or persistent paresthesias (95% confidence interval, 0%-0.08%). In one patient, the epidural catheter broke during removal, and a portion was retained. The patient was notified; no long term sequelae were noted. Six patients developed new neurologic symptoms or postoperative worsening of a previous neurologic condition unrelated to epidural catheterization. We conclude that the risk of neurologic complications associated with lumbar epidural catheter placement in anesthetized patients is small. However, the relative risk of this practice, compared with epidural catheter placement in awake patients, is unknown. IMPLICATIONS: We report no neurologic complications in 4298 patients undergoing epidural catheter placement while under general anesthesia. Although the risk of neurologic complications associated with lumbar epidural catheter placement in anesthetized patients is small, the relative risk compared with epidural catheterization in awake patients is unknown. PMID- 12760974 TI - The effects of epidural and general anesthesia on tissue oxygenation. AB - The risk of wound infections is inversely related to subcutaneous tissue oxygen tension. General anesthesia increases local blood flow by direct vasodilation and central inhibition of thermoregulatory vasoconstriction. Epidural anesthesia can increase perfusion in blocked regions by decreasing sympathetic tone. We therefore tested the hypothesis that epidural anesthesia increases tissue oxygen tension in awake and anesthetized subjects. Fifteen healthy volunteers underwent epidural, general, and combined epidural and general anesthesia. Subcutaneous tissue oxygen tension was measured using tonometers in the lateral upper arm and the lateral thigh. Epidural anesthesia to a T10 level was maintained with 0.75% mepivacaine. General anesthesia was maintained with 1.5% sevoflurane in 30% oxygen; 30% inspired oxygen was given via a sealed facemask during baseline and epidural anesthesia. Baseline subcutaneous tissue oxygen tensions for arm and thigh were 57 +/- 11 and 54 +/- 8 mm Hg, respectively. Epidural anesthesia significantly increased tissue oxygenation in the thigh by 9 mm Hg, to 63 +/- 7 mm Hg, without increasing arm oxygenation. Tissue oxygenation in the arm and thigh were similar during general anesthesia alone, 58 +/- 11 and 63 +/- 12 mm Hg. Arm oxygenation remained unchanged with the addition of epidural anesthesia; however, thigh subcutaneous oxygen partial pressure increased 8 +/- 3 mm Hg, from 63 +/- 12 to 71 +/- 9 mm Hg. Although epidural anesthesia increased tissue oxygenation significantly with and without general anesthesia, the magnitude of this increase might be of marginal clinical importance in regard to surgical wound infections. IMPLICATIONS: Epidural anesthesia significantly increased subcutaneous tissue oxygenation in the thigh both with and without general anesthesia. Although each increase was statistically significant, previous work suggests that the magnitude of these changes is unlikely to markedly reduce the risk of surgical wound infection. PMID- 12760975 TI - Knowledge and practice regarding prophylactic perioperative beta blockade in patients undergoing noncardiac surgery: a survey of Canadian anesthesiologists. AB - A lack of awareness of the "best" current practice is frequently cited as a major barrier to the practice of evidence-based medicine. The purpose of this study was to survey Canadian anesthesiologists to determine their knowledge and practices associated with prophylactic perioperative beta blockade, a therapy that has been widely discussed in the literature and has the potential for a significant positive impact on patient outcomes. We sent questionnaires to 1234 members of the Canadian Anesthesiologists' Society. The overall response rate was 54%. Ninety-five percent of respondents were aware of the perioperative beta blocker literature, and of these, 93% agreed that beta blockers were beneficial in patients with known coronary artery disease (CAD). Fifty-seven percent reported always or usually administering prophylactic beta blockers in patients with known CAD, and 34% of these regular users continued therapy beyond the early postoperative period. Only 9% of respondents reported that a formal protocol existed at their facility. This study suggests that barriers to the translation of research to practice were not related to a lack of awareness of the current best evidence. With respect to perioperative beta blockers, controversies within the literature as well as practical considerations may be greater barriers to implementation of best evidence. IMPLICATIONS: This survey found that anesthesiologists were aware of and supported the use of prophylactic perioperative beta blockers in patients with risk factors or known coronary artery disease; however, only 57% frequently prescribed perioperative beta blockers. A lack of awareness of the current "best" evidence was not a barrier to use. PMID- 12760976 TI - Perioperative myocardial ischemia in patients undergoing sternectomy shortly after coronary artery bypass grafting. AB - Coronary revascularization reduces cardiac complications associated with noncardiac surgery in patients with severe coronary disease. However, patients undergoing emergency noncardiac surgery soon after coronary bypass operations may still be vulnerable to ischemic myocardial events. We prospectively evaluated the incidence of myocardial ischemia in 82 consecutive patents scheduled for sternectomy in the first (Group 1; 35 patients) or second (Group 2; 47 patients) week after coronary artery bypass graft (CABG) surgery. The interval between CABG surgery and sternectomy in Groups 1 and 2 was 6 days (range, 4-7 days) and 11 days (range, 8-14 days), respectively. Electrocardiographic (ECG) changes consistent with myocardial ischemia were assessed with a two-channel Holter system for 48 h. There were no between-group differences in updated Acute Physiology and Chronic Health Evaluation score, use of beta-blockers, or perioperative hemodynamic changes. The incidence of ECG changes consistent with myocardial ischemia was fivefold more frequent in Group 1 (22.85% versus 4.25%; P < 0.05). Of the ischemic patients in Group 1, 25% experienced a perioperative acute myocardial infarction (one was fatal). There were no infarcts in Group 2. Thus, patients appear to be prone to coronary events during sternectomy performed early after CABG surgery. Although the incidence of ischemia did not differ from that previously reported after CABG surgery alone, further investigation is required to determine whether the findings obtained in this high-risk population are generalizable to patients undergoing noncardiac surgery soon after uneventful CABG surgery. IMPLICATIONS: This study demonstrates an increased incidence of myocardial ischemia when sternectomy for mediastinitis is performed within one week of coronary artery bypass graft surgery, and this ischemia is associated with a 25% incidence of myocardial infarction. PMID- 12760977 TI - Acute intravascular volume expansion with rapidly administered crystalloid or colloid in the setting of moderate hypovolemia. AB - Although the distribution of various crystalloid and colloid solutions at equilibrium has been well established, the acute peak expansion of intravascular volume that can be achieved with the rapid administration of crystalloid or colloid is unknown. We studied eight healthy male subjects in a two-part crossover trial designed to assess the maximal increase in intravascular volume achieved with 1000 mL of lactated Ringer's solution compared with the same volume of 6% Hetastarch. Subjects were made moderately hypovolemic by the withdrawal of 900 mL of blood, and then the crystalloid or colloid solution was rapidly infused over 5-7 min. Serial dilution of hematocrit was measured every 5 min for 30 min to determine changes in blood volume. Peak expansion of intravascular volume with lactated Ringer's solution was 630 +/- 127 mL, occurring immediately the rapid infusion was complete, whereas the peak expansion of intravascular volume with 6% Hetastarch was 1123 +/- 116 mL and occurred 5 min after the completion of the fluid infusion. The results were significantly different (P < 0.001). These results would suggest that even for very short periods of time, rapid infusion of colloid significantly more effectively increases blood volume and, by inference, cardiac output than the same volume of crystalloid, even if the crystalloid is administered very rapidly. IMPLICATIONS: Under conditions of moderate hypovolemia, the maximal acute intravascular volume expansion with the rapid infusion of 1000 mL of lactated Ringer's solution is slightly more than half that achieved with the same volume of 6% Hetastarch. PMID- 12760978 TI - The effect of dexamethasone on side effects after coronary revascularization procedures. AB - Corticosteroids decrease side effects after noncardiac elective surgery. We designed this randomized, double-blinded, placebo-controlled study to test the hypothesis that standard doses of dexamethasone (4 mg x2) would reduce postoperative nausea, vomiting, and pain, decrease the incidence of atrial fibrillation (AF), and improve appetite after cardiac surgery, thereby facilitating the recovery process. A total of 300 patients undergoing coronary revascularization surgery were enrolled in this clinical study. The anesthetic management was standardized in all patients. Dexamethasone (4 mg/mL) or saline (1 mL) was administered after the induction of anesthesia and a second dose of the same study drug was given on the morning after surgery. The incidence of AF was determined by analyzing the first 72 h of continuously recorded electrocardiogram records after cardiac surgery. The patients were assessed at 24- and 48-h intervals after surgery, as well as at the time of hospital discharge, to determine the incidence and severity of postoperative side effects (e.g., nausea, vomiting, pain) and patient satisfaction scores. Dexamethasone significantly reduced the need for antiemetic rescue medication on the first postoperative day (30% versus 42%), and the incidences of nausea (15% versus 26%) and vomiting (5% versus 16%) on the second postoperative day (P < 0.05). In addition, dexamethasone significantly reduced the percentage of patients with a depressed appetite on the second postoperative day. However, the corticosteroid failed to decrease the incidence of AF (27% versus 32%) or the total dosage of opioid analgesic medication administered in the postoperative period. We conclude that dexamethasone (8 mg in divided doses) was beneficial in reducing emetic symptoms and improving appetite after cardiac surgery. However, this dose of the corticosteroid does not seem to have antiarrhythmic or analgesic-sparing properties. IMPLICATIONS: Dexamethasone (8 mg IV) was beneficial in reducing emetic symptoms and increasing appetite after cardiac surgery. However, this dose of the corticosteroid failed to decrease postoperative pain or the incidence of new-onset atrial fibrillation. PMID- 12760979 TI - Supplementing transesophageal echocardiography with transthoracic echocardiography for monitoring transcatheter closure of atrial septal defects with attenuated anterior rim: a case series. AB - The use of transesophageal echocardiography (TEE) for guidance of transcatheter closure of secundum-type atrial septal defect (ASD) is increasingly becoming a routine procedure. ASD with attenuated anterior superior (SA) rim is a variant of secundum-type ASD and is suitable for transcatheter closure. The success rate of TEE guidance for device deployment in these patients is not known. Therefore, we assessed 124 consecutive patients with ASD (57 secundum-type, 67 with attenuated SA rim) closed with an Amplatzer Septal Occluder under TEE guidance. Our results show that the TEE was successful in depicting all 4 corners and corresponding edges of each Amplatzer disk, as well as the septal rims of all 57 secundum-type ASDs. However, in 6 of 67 ASDs (9%) with attenuated SA rim in which TEE failed to visualize the adequate placement of occluder on the anterior inferior (IA) rim, the additional use of transthoracic echocardiography helped to resolve this inadequacy. Four of these six patients had the unusual morphology of the IA rim tissue. Two had severe right axis deviation of the heart with large Q angle (>90 degrees ). The SA rim was absent in 35 of 67 ASDs with attenuated SA rim and in these cases TEE demonstrated the anterior surface of the disk against the wall of the aorta but without distortion. We conclude that TEE can be useful for confirming successful deployment of the occluder in most patients with ASDs. In a small number of ASDs with attenuated SA rim who have unusual IA morphology, supplemental transthoracic echocardiography is required to verify successful deployment of the occluder when TEE visualization fails to reliably diagnose adequate placement of the occluder. IMPLICATIONS: Transesophageal echocardiography can be useful for confirming successful deployment of the occluder in the majority of patients with atrial septal defect. In a small number of atrial septal defects with attenuated anterior superior rim which have unusual anterior inferior morphology, supplemental transthoracic echocardiography is required to verify successful deployment of the occluder when transesophageal echocardiography visualization fails to reliably diagnose adequate placement of the occluder. PMID- 12760980 TI - The contribution of the coronary concentrations of propofol to its cardiovascular effects in anesthetized sheep. AB - Linking physiological pharmacokinetic models to models of the cardiovascular system requires knowledge of the sites in the body that mediate a drug's cardiovascular effects. We examined the role of the coronary concentrations of propofol. Nine sheep anesthetized with isoflurane (2%) were instrumented acutely for cardiovascular measurements. In a random crossover design, they were administered ramped coronary artery (CA) infusions of propofol to selectively enrich the myocardium (as indicated by the coronary sinus blood concentration) or IV infusions to achieve the same concentration range in all sites of the body. Reductions in left ventricular myocardial contractility (LV dP/dt(max)) and mean arterial blood pressure were linearly related to the propofol concentration. For the CA route, LV dP/dt(max) was reduced by 52 mm Hg/s for each milligram per liter increase in coronary sinus propofol concentration. For the IV route, the reduction in LV dP/dt(max) was equivalent to that with the CA route, showing that the coronary propofol concentration was the major contribution to this effect. For the CA route, mean arterial blood pressure was reduced by 0.6 mm Hg for each milligram per liter. There was a larger reduction (2.5 mm Hg x mg(-1) x L(-1)) for the IV route. Therefore, this effect was predominantly mediated by propofol concentrations elsewhere in the body. IMPLICATIONS: With use of selective coronary artery infusions in sheep, the coronary concentrations of propofol were shown to be the major contributor to the cardiac depression caused by propofol but were a less significant contributor to the hypotension caused by this drug. Models of the cardiovascular effects of propofol should account for these relationships. PMID- 12760981 TI - Right ventricular exclusion surgery for arrhythmogenic right ventricular dysplasia with cardiomyopathy. AB - IMPLICATIONS: The authors describe the management of a patient with arrhythmogenic right ventricular dysplasia treated with right ventricular exclusion surgery. PMID- 12760982 TI - Management of pulmonary hypertension: physiological and pharmacological considerations for anesthesiologists. PMID- 12760983 TI - The effects of magnesium prime solution on magnesium levels and potassium loss in open heart surgery. AB - In this study, we examined the effects of magnesium supplementation in the cardiopulmonary bypass (CPB) prime solution on pediatric patients' magnesium levels and potassium loss with open heart surgery. Forty pediatric patients undergoing heart surgery were randomly assigned either magnesium sulfate (magnesium group, n = 20; 0.25 mmol/kg) or saline (placebo group; n = 20) supplementation to the prime solution. Ionized magnesium (IMg) and urinary magnesium and potassium were measured at defined time points during and after CPB. In the magnesium group, IMg concentration was larger during CPB but not after CPB. IMg decreased in the early stages of CPB in the placebo group and decreased to an even smaller level 24 h after CPB. Urinary magnesium levels in the magnesium group were larger than those in the placebo group during and after CPB, and urinary potassium concentrations reached significantly smaller levels 24 h after CPB (44.2 +/- 2.9 versus 60.9 +/- 2.6 mmol/L; P < 0.01). We conclude that the addition of magnesium into prime solution maintains normal IMg levels and prevents potassium flux during the perioperative period. IMPLICATIONS: In our study, we demonstrate that a magnesium prime solution can prevent hypomagnesemia during and after cardiopulmonary bypass (CPB) and decrease the urinary potassium loss after CPB in pediatric patients undergoing open heart surgery. PMID- 12760984 TI - Postoperative analgesia in children undergoing myringotomy and placement equalization tubes in ambulatory surgery. AB - We enrolled 120 children undergoing bilateral myringotomy and tube placement in this prospective, randomized, observer-blinded study. Patients were randomized into one of four groups: Group 1 (control) was plain acetaminophen 10 mg/kg orally, Group 2 was acetaminophen 10 mg/kg with 1 mg/kg of codeine orally, Group 3 was transnasal butorphanol 25 micro g/kg given immediately after the induction of anesthesia, and Group 4 was ketorolac 1 mg/kg given IM immediately after the induction of anesthesia. All children received oral midazolam (0.6 mg/kg) before surgery. A nurse blinded to the analgesic technique used assessed the child's behavior at the induction of anesthesia and in the postanesthesia care unit using a 4-point scale. Analgesic effectiveness was determined by assessing the child's pain at 5-min intervals using a modified 10-point objective pain scale. In the postanesthesia care unit, rescue pain medication was administered for an objective pain scale >or=4 or a behavior score >or=3. Our data suggest that IM ketorolac is a promising analgesic to be used in this surgical population. Time to first rescue analgesic was longest in the ketorolac group, and there was no associated postoperative vomiting or nausea. IM ketorolac given during surgery was the best analgesic regimen for these procedures. IMPLICATIONS: We compared four different analgesics in the management of pain after placement of pressure equalization tubes during myringotomy in children and demonstrated that ketorolac or butorphanol provided superior analgesia when compared with acetaminophen with codeine or plain acetaminophen. Children who received ketorolac versus butorphanol had less vomiting in the 24 h after surgery. PMID- 12760985 TI - A prospective cohort study of emergence agitation in the pediatric postanesthesia care unit. AB - Emergence agitation (EA) is a postanesthetic problem that interferes with a child's recovery and presents a challenge in terms of assessment and management. In this prospective cohort study, we sought to determine the incidence of EA, evaluate factors associated with and predictive of EA, and describe associated outcomes in healthy children. Children aged 3-7 yr who were undergoing general anesthesia for elective outpatient procedures were included. All perioperative care was documented, and postoperative behaviors in the postanesthesia care unit were recorded. Parents completed the Behavioral Style Questionnaire for 3- to 7 yr-olds. Five-hundred-twenty-one children were studied, of whom 96 (18%) had EA. Agitation lasted up to 45 min in some cases (range, 3-45 min; mean, 14 +/- 11 min), required pharmacologic intervention in 52% of children, and was associated with a prolonged postanesthesia care unit stay (117 +/- 66 min versus 101 +/- 61 min for nonagitated children; P = 0.02). Ten factors were found to be associated with EA, including age, previous surgery, adaptability, ophthalmology and otorhinolaryngology procedures, sevoflurane, isoflurane, sevoflurane/isoflurane, analgesics, and time to awakening. Of these, otorhinolaryngology procedures, time to awakening, and isoflurane were shown to be independent risk factors. IMPLICATIONS: Children may become agitated after general anesthesia. This study describes several factors that may increase the risk for agitation. These data are important in planning anesthesia care for young children. PMID- 12760986 TI - The efficacy of celecoxib premedication on postoperative pain and recovery times after ambulatory surgery: a dose-ranging study. AB - Recently, the Food and Drug Administration increased the celecoxib dosage recommendation from 200 mg to 400 mg for acute pain management. No studies have directly compared the analgesic efficacy of different doses of celecoxib for the prevention of postoperative pain. In this prospective, double-blinded, placebo controlled study, we compared oral celecoxib 200 mg to 400 mg when administered for premedication of outpatients undergoing minor ear-nose-throat surgery. A total of 93 healthy outpatients were assigned to 1 of 3 study groups: control (placebo; n = 30), celecoxib 200 mg (n = 30), or celecoxib 400 mg (n = 33). The study drug was given orally 30-45 min before surgery, and all patients received a standardized general anesthetic technique. During the postoperative period, pain scores (0-10), recovery times, the need for rescue analgesics, quality of recovery (0-100), patient satisfaction with pain management (0-100), and side effects were recorded. Pain was assessed at 30-min intervals using a verbal rating scale, with 0 = no pain to 10 = worst pain imaginable, in the postanesthesia care unit and day surgery unit recovery areas and at 24 h after surgery. Celecoxib 400 mg was significantly more effective than 200 mg (and placebo) in reducing postoperative pain. Both celecoxib 200 mg and 400 mg were more effective than placebo in reducing the postoperative fentanyl requirement (74 +/- 67 micro g and 56 +/- 62 micro g versus 120 +/- 86 micro g, respectively). The larger dose of celecoxib significantly reduced the percentage of patients with severe pain at discharge (6% versus 37% and 30% in the celecoxib 200 mg and control groups, respectively). The median number of doses of oral analgesic medication after discharge was also significantly reduced in the celecoxib 400 mg group (0 versus 2 and 2 in the celecoxib 200 mg and control groups, respectively). However, no differences were found among the three study groups with respect to recovery times and secondary outcome variables (e.g., patient satisfaction and quality of recovery). We conclude that oral premedication with celecoxib 400 mg was more effective than 200 mg in reducing severe postoperative pain and the need for rescue analgesic medication in the postoperative period. IMPLICATIONS: Oral premedication with celecoxib 400 mg was more effective than 200 mg in reducing postoperative pain and the need for rescue analgesic medication in the early postoperative period. However, neither dose of celecoxib was more effective than a placebo in facilitating the recovery process after outpatient surgery. PMID- 12760987 TI - Can the bispectral index be used to predict seizure time and awakening after electroconvulsive therapy? AB - The electroencephalogram (EEG) bispectral index (BIS) measures the hypnotic component of the anesthetic state and correlates with emergence from general anesthesia. Therefore, we hypothesized that the BIS would be useful in predicting electroconvulsive therapy (ECT)-induced seizure times and awakening from methohexital anesthesia. Twenty-five consenting patients with major depressive disorders underwent 100 maintenance ECT treatments. All patients were premedicated with glycopyrrolate 0.2 mg IV, and anesthesia was induced with methohexital 1 mg/kg IV. The BIS was monitored continuously, and the values were recorded at specific end-points, including before anesthesia (baseline), after the induction of anesthesia (pre-ECT), at the end of ECT (peak), after ECT (suppression), and on awakening (eye opening). The pre-ECT BIS value correlated with the duration of both the motor (r = 0.3) and EEG (r = 0.4) seizure activity (P < 0.05). The peak post-ECT BIS value correlated with the duration of the EEG seizure activity (r = 0.5) (P < 0.05). A positive correlation was also found between the EEG seizure duration and the time to eye opening (r = 0.4) (P < 0.05). However, the BIS values on awakening from methohexital anesthesia varied from 29 to 97 and were <60 in 75% of the cases. We conclude that the BIS value before the ECT stimulus is applied could be useful in predicting the seizure time. However, the BIS values on awakening were highly variable, suggesting that it reflects both the residual depressant effects of methohexital and post-ictal depression. IMPLICATIONS: The bispectral index (BIS) value immediately before the electroconvulsive therapy (ECT) stimulus correlates with the duration of the motor and electroencephalogram (EEG) seizure activity during methohexital anesthesia. In addition, the increase in the BIS value during the ECT-induced seizure was proportional to the duration of EEG seizure activity. However, the BIS value on awakening from anesthesia varied widely, from 29 to 97. PMID- 12760988 TI - Postoperative myalgia after succinylcholine: no evidence for an inflammatory origin. AB - A common side effect associated with succinylcholine is postoperative myalgia. The pathogenesis of this myalgia is still unclear; inflammation has been suggested but without convincing evidence. We designed the present study to investigate whether an inflammatory reaction contributes to this myalgia. The incidence and severity of succinylcholine-associated myalgia was determined in 64 patients pretreated with saline or dexamethasone before succinylcholine (n = 32 for each). Incidence and severity of myalgia did not differ significantly between the two groups: 15 patients in the dexamethasone group complained of myalgia compared with 18 patients in the saline group, and severe myalgia was reported by five patients and three patients, respectively (not significant). At 48 h after surgery, 12 patients in both groups still suffered from myalgia (not significant). In addition, interleukin-6 (IL-6) as an early marker of inflammation was assessed in a subgroup of 10 patients pretreated with saline. We found an increase of IL-6 for only three patients, but only one patient reported myalgia; no relationship between myalgia and the increase of IL-6 was found. In conclusion, there is no evidence for an inflammatory origin of succinylcholine associated myalgia. IMPLICATIONS: Administration of dexamethasone before succinylcholine was not effective in decreasing the incidence or the severity of succinylcholine-induced postoperative myalgia. Furthermore, there was no significant relationship between postoperative myalgia and time course of interleukin-6 concentrations, a marker of inflammation. Pretreatment with dexamethasone is not justified to prevent postoperative myalgia after succinylcholine. PMID- 12760989 TI - Sympathetic and vascular consequences from remifentanil in humans. AB - We explored the possible mechanisms of hypotension during the administration of sedation-analgesia doses of remifentanil in young (ASA physical status I) volunteers (n = 24). Cardiorespiratory and sympathetic variables were collected at baseline and at plasma concentrations of remifentanil (2 and 4 ng/mL). Monitoring included electrocardiogram, heart rate (HR), direct blood pressure, muscle sympathetic nerve activity, and forearm blood flow (FBF). A cold pressor test (1-min hand immersion in ice water) quantified analgesia effectiveness (visual analog scale, 0-100). Visual analog scale to the cold pressor test (62 at baseline) decreased to 27 and 18 during remifentanil infusions. Respiratory rate decreased and end-tidal carbon dioxide (ETCO(2)) increased with increasing doses of remifentanil; HR, direct blood pressure, muscle sympathetic nerve activity, SpO(2) remained unchanged, but FBF increased compared with placebo. In a second study (n = 7), timed respiration was used to maintain ETCO(2) during remifentanil, but FBF still increased. In a third study (n = 11), direct effects of remifentanil on vascular tone were determined with progressive infusions from 1 to 100 micro g/h into the brachial artery; FBF increased significantly from 3.5 to 4.3 mL/min per 100 mL of tissue (approximately 13%-18% increase). Sedative doses of remifentanil resulted in analgesia but no changes in neurocirculatory end-points except FBF. Direct effects of remifentanil on regional vascular tone may play a role in promoting hypotension. IMPLICATIONS: Remifentanil occasionally has been associated with hypotension, the mechanism of which is unclear. This study found that remifentanil directly causes the forearm arterial vasculature to dilate. PMID- 12760990 TI - Effect-site concentration of propofol for recovery of consciousness is virtually independent of fentanyl effect-site concentration. AB - Fentanyl reduces the amount of propofol necessary to prevent responses to surgical stimuli. However, opioids have relatively little effect on consciousness. We, therefore, tested the hypothesis that fentanyl minimally alters the effect-site concentration of propofol associated with awakening. Fifty women having gynecologic laparotomy with propofol anesthesia were randomly allocated into the following target effect-site fentanyl concentrations: 0.8, 1.0, 1.4, 2.0, and 3.0 ng/mL. Fentanyl was continued at the designated rate through the initial postoperative phase. The propofol effect-site concentration associated with eye opening in response to verbal command was regarded as the awakening concentration. The estimated propofol effect-site concentrations at awakening did not differ significantly among the groups and were 1.9 +/- 0.5 micro g/mL with a fentanyl effect-site concentration of 0.8 ng/mL; 1.6 +/- 0.4 micro g/mL with 1.0 ng/mL of fentanyl; 1.6 +/- 0.2 micro g/mL with 1.4 ng/mL of fentanyl; 1.7 +/- 0.4 micro g/mL with 2.0 ng/mL of fentanyl; and 1.6 +/- 0.34 micro g/mL with 3.0 ng/mL of fentanyl (mean +/- SD). Seventy percent of the subjects in the 0.8 ng/mL fentanyl group spontaneously complained of pain, whereas none of the patients in the 2 or 3 ng/mL groups did. Five (56%) of 9 women in the 3 ng/mL group had a postoperative respiratory rate <6 breaths/min. Heart rate in one of these women decreased to <40 bpm. These data suggest that the optimal fentanyl effect-site concentration in patients recovering from gynecologic laparoscopy is between 1.4 and 2.0 ng/mL. IMPLICATIONS: The effect site concentration for propofol at awakening was virtually independent of the fentanyl effect-site concentration over the range of 0.8 to 3.0 ng/mL; however, 0.8 ng/mL of fentanyl was associated with inadequate postoperative analgesia, and 3.0 ng/mL of fentanyl was associated with respiratory toxicity. The optimal postoperative fentanyl effect-site concentration during recovery from propofol general anesthesia for laparotomy thus appears to be near 2 ng/mL. PMID- 12760991 TI - The effects of local and intravenous anesthetics on recombinant rat VR1 vanilloid receptors. AB - Capsaicin, acting at the vanilloid 1 receptor (VR1), may potentiate local anesthetic activity, and as a ligand-gated ion channel of the transient receptor potential family, may also be a target for IV general anesthetics. We have examined whether local (lidocaine, prilocaine, and procaine 0.1-10 mM; 10 mM represents 0.25%-0.27% wt/vol) or IV anesthetics (propofol 10 micro M, thiopental 100 micro M, and ketamine 100 micro M) interact with recombinant rat VR1 expressed in human embryonic kidney (HEK293) cells (VR1-HEK293). We have assessed receptor interaction functionally by monitoring intracellular Ca(2+) ([Ca(2+)](i)) in Fura2-loaded cells at 37 degrees C. The addition of capsaicin (60 nM) produced a time-dependent biphasic increase in [Ca(2+)](i) amounting to 50-100 nM above than basal, which was inhibited by capsazepine 10 micro M and was absent in wild type HEK293 cells. Lidocaine and prilocaine alone (e.g., at 10 mM) significantly increased [Ca(2+)](i) by 67 +/- 6 nM and 33 +/- 7 nM, respectively, and concentration-dependently inhibited the capsaicin response. The effects of procaine were obscured by anesthetic-induced quenching of Fura2. In wild type HEK293 cells, lidocaine (10 mM) alone produced a small increase in [Ca(2+)](i). All IV anesthetics failed to modify capsaicin-increased [Ca(2+)](i). In conclusion, the present data suggest that local but not IV general anesthetics interact with recombinant rat VR1 receptors with the former anesthetics having antagonistic activity. IMPLICATIONS: Vanilloid receptors (VR1) are activated by capsaicin, the pain-producing component of hot chili peppers. We suggest that local (but not IV general) anesthetics may have inhibitory actions on this receptor. PMID- 12760992 TI - Alpha-2 adrenoreceptors probably do not mediate the immobility produced by inhaled anesthetics. AB - Agonism of alpha-adrenoreceptors has a powerful anesthetic result mediated, in part, by effects on the spinal cord. Alpha-adrenoreceptor agonists (e.g., dexmedetomidine) can decrease the minimum alveolar anesthetic concentration (MAC) of inhaled anesthetics (e.g., halothane) to zero, with an apparently additive interaction between halothane and dexmedetomidine. We tested whether the capacity of the inhaled anesthetic isoflurane to produce immobility in the face of noxious stimulation resulted from agonism of alpha-adrenoreceptors. MAC (the concentration required to eliminate movement in response to a noxious stimulus in 50% of subjects) of isoflurane was determined before and after intraperitoneal administration of the alpha-adrenoreceptor antagonists yohimbine and atipamezole. The doses of yohimbine and atipamezole equaled or exceeded those that reverse the ability of agonism of alpha-adrenoreceptors to decrease MAC. Smaller doses of yohimbine or atipamezole slightly increased (by 10%) the MAC of isoflurane, an increase we interpret as the result of blockade of a small amount of tonically active alpha-adrenoreceptor activity. Doses five-fold larger did not change MAC. Doses 10-fold larger decreased MAC. We conclude that alpha-adrenoreceptors do not or minimally mediate the capacity of inhaled anesthetics to produce immobility. IMPLICATIONS: Although stimulation (agonism) of alpha-2 adrenoreceptors can decrease the inhaled anesthetic concentration required to produce immobility in the face of noxious stimulation, blockade of alpha-2 adrenoreceptors minimally affects the concentration. Thus, augmentation of the effect of alpha-2 adrenoreceptors is not an appreciable part of the mechanism whereby inhaled anesthetics produce immobility. PMID- 12760993 TI - Inhibition of human TREK-1 channels by bupivacaine. AB - Human TWIK-related K(+) channels (TREK-1) stabilize the membrane potential (mp) of neurons and have a major role in the regulation of membrane excitability. In view of their physiological significance, interaction of bupivacaine with TREK-1 channels may be clinically important. Our aim was to characterize with the patch clamp technique the properties of human TREK-1 channels and the effects of bupivacaine on these channels expressed in Chinese hamster ovary (CHO) cells. Transfection of CHO cells with TREK-1 channels (CHO(TREK-1) cells) hyperpolarized the mp from -33 +/- 13 to -78 +/- 4 mV. The channels were stimulated by intracellular acidosis. Inhibition of TREK-1 channels by bupivacaine was reversible, concentration-dependent, voltage-independent, and increased with intracellular acidosis. Bupivacaine depolarized the mp of CHO(TREK-1) cells in a reversible and concentration-dependent manner. Concentrations for channel inhibition and membrane depolarization were not linearly related (50% inhibitory concentration value for channel inhibition 370 +/- 20 micro M, Hill coefficient 1.8 +/- 0.1, n = 51; 50% inhibitory concentration value for membrane depolarization 856 +/- 14 micro M, Hill coefficient 2.4 +/- 0.1, mean +/- SEM, n = 27). The results suggest that protonated bupivacaine elicits the observed effects via a site of interaction accessible from the intracellular space. Inhibition of TREK-1 channels and consecutive depolarization of the cell membrane by bupivacaine may contribute to blockade of neuronal signal conduction during regional anesthesia. IMPLICATIONS: The interaction of bupivacaine with human TREK 1 channels was studied with the patch-clamp technique. Bupivacaine inhibited TREK 1 channels and depolarized the membrane potential of cells expressing TREK-1 channels in a concentration-dependent and reversible manner. Both effects may contribute to conductance block caused by bupivacaine. PMID- 12760995 TI - Olprinone for the treatment, but not prevention, of fatigue-induced changes in guinea-pig diaphragmatic contractility. AB - Olprinone, a phosphodiesterase III inhibitor, improves the contractility in fatigued diaphragm in vivo, but no data are available for the treatment and prevention of fatigue-induced changes in vitro. We therefore examined the efficacy of Olprinone for the treatment and prevention of fatigue-induced changes in guinea-pig diaphragmatic contractility. The guinea-pig diaphragm strips were randomly allocated according to dose of Olprinone (0, 10(-6), 10(-5), and 10(-4) M) (n = 7 each) and were stimulated directly in an organ bath. Diaphragmatic contractility was measured by assessing twitch tension and force at 20-Hz and 100 Hz stimulation. Diaphragmatic fatigue was induced by generating rhythmic, repetitive contractions produced by 20-Hz stimulation for 5 min. In the first experiment, after the fatigue-producing period, Olprinone was administered to the organ bath for 5 min. In the second experiment, Olprinone was pretreated for 5 min, and then diaphragmatic fatigue was produced. In Experiment 1, after a fatigue-producing period, tetanic force to each stimulus decreased from baseline values (P < 0.05). Olprinone 10(-5)-10(-4) M caused an increase in force at both stimuli from fatigued values (P < 0.05). In Experiment 2, no change in tetanic force was observed by pretreatment with Olprinone (0-10(-4) M). After producing fatigue, tetanic force to each stimulus decreased from baseline values (P < 0.05). These results suggest that Olprinone 10(-5)-10(-4) M improves the fatigue induced changes in guinea-pig diaphragmatic contractility and that pretreatment with Olprinone does not prevent diaphragmatic fatigability. IMPLICATIONS: Olprinone is effective for the treatment, but not prevention, of fatigue-induced changes in guinea-pig diaphragmatic contractility. PMID- 12760994 TI - The effects of enflurane, isoflurane, and intravenous anesthetics on rat diaphragmatic function and fatigability. AB - We examined the effect of isoflurane, enflurane, midazolam, ketamine, propofol, and thiopental on diaphragmatic functions under unfatigued and fatigued conditions in 228 rat isolated muscle strips. Diaphragmatic twitch characteristics and tetanic contractions were measured before and after muscle fatigue, which was induced by repetitive tetanic contraction with or without exposure to one of the anesthetics at clinically relevant plasma concentrations, and at 10 and 100 times this concentration, or at 1, 2, and 3 minimum alveolar anesthetic concentration (MAC). Isoflurane, midazolam, ketamine, propofol, and thiopental did not induce a direct inotropic or lusitropic effect under unfatigued and fatigued conditions. Enflurane did not change contraction or relaxation in fresh isolated diaphragm, but enflurane at 2-3 MAC enhanced diaphragmatic fatigability itself and fatigue-induced impairment of twitch characteristics and tetanic tensions. These effects were greater at 3 MAC than at 2 MAC. Our findings suggest that the reduction of diaphragm function previously reported in in vivo experiments using propofol, midazolam, and isoflurane is not related to a direct effect on intrinsic diaphragmatic contractility. Our results also indicate that large concentrations of enflurane may impair the diaphragmatic function at sites other than excitation-contraction coupling. IMPLICATIONS: Enflurane did not change contraction or relaxation in fresh isolated rat diaphragm, but enhanced diaphragmatic fatigability itself and fatigue-induced impairment of twitch characteristics and tetanic tensions. Isoflurane, midazolam, ketamine, propofol, and thiopental had no direct effects on diaphragmatic functions under unfatigued and fatigued conditions. Isoflurane and these i.v. anesthetics may be advantageous over enflurane to anesthetize and/or sedate patients who are predisposed to diaphragmatic fatigue. PMID- 12760996 TI - Resistive-heating and forced-air warming are comparably effective. AB - Serious adverse outcomes from perioperative hypothermia are well documented. Consequently, intraoperative warming has become routine. We thus evaluated the efficacy of a novel, nondisposable carbon-fiber resistive-heating system. Twenty four patients undergoing open abdominal surgery lasting approximately 4 h were randomly assigned to warming with 1) a full-length circulating water mattress set at 42 degrees C, 2) a lower-body forced-air cover with the blower set on high, or 3) a three-extremity carbon-fiber resistive-heating blanket set to 42 degrees C. Patients were anesthetized with a combination of continuous epidural and general anesthesia. All fluids were warmed to 37 degrees C, and ambient temperature was kept near 22 degrees C. Core (tympanic membrane) temperature changes among the groups were compared by using factorial analysis of variance and Scheffe F tests; results are presented as means +/- SD. Potential confounding factors did not differ significantly among the groups. In the first 2 h of surgery, core temperature decreased by 1.9 degrees C +/- 0.5 degrees C in the circulating-water group, 1.0 degrees C +/- 0.6 degrees C in the forced-air group, and 0.8 degrees C +/- 0.2 degrees C in the resistive-heating group. At the end of surgery, the decreases were 2.0 degrees C +/- 0.8 degrees C in the circulating-water group, 0.6 degrees C +/- 1.0 degrees C in the forced-air group, and 0.5 degrees C +/- 0.4 degrees C in the resistive-heating group. Core temperature decreases were significantly greater in the circulating-water group at all times after 150 elapsed minutes; however, temperature changes in the forced-air and resistive heating groups never differed significantly. Even during major abdominal surgery, resistive heating maintains core temperature as effectively as forced air. IMPLICATIONS: Efficacy was similar for forced-air and resistive heating, and both maintained intraoperative core temperature far better than circulating-water mattresses. We thus conclude that even during major abdominal surgery, resistive heating maintains core temperature as effectively as forced air. PMID- 12760997 TI - Core temperature monitoring with new ventilatory devices. AB - Widespread use of new airway devices, such as the laryngeal mask airway (LMA) and the cuffed oropharyngeal airway (COPA), preclude measuring core temperature in the distal esophagus. Therefore, we tested the hypothesis that core temperature measured with a thermocouple positioned on a LMA or COPA is sufficiently accurate and precise for clinical use. Temperatures were recorded from thermocouples positioned on the cuffs of LMAs or COPAs in 36 patients scheduled for prolonged orthopedic surgery or therapeutic hyperthermia for cancer. These temperatures, recorded at 15-min intervals, were compared with simultaneously obtained nasopharynx and tympanic membrane temperatures. Data were compared by linear regression and the bias calculated. Temperatures measured on the LMA correlated well with both nasopharyngeal (r(2) = 0.94) and tympanic membrane (r(2) = 0.94) temperatures. Temperatures measured on the COPA also correlated well with those on the nasopharynx (r(2) = 0.97) and tympanic membrane (r(2) = 0.96). The fraction of temperatures that differed from nasopharynx temperature by more than +/-0.5 degrees C was 8% with LMA and 11% with COPA; the fraction of temperatures that differed from tympanic temperature by more than +/-0.5 degrees C was 7% with LMA and 10% with COPA. These results suggest that body temperature measured from the cuffs of COPA or LMAs is sufficiently accurate for routine clinical use. IMPLICATIONS: Temperatures measured on airway devices correlated well with independent measurements of core body temperature. Thus, body temperature measured on the cuffs of airway devices is sufficiently accurate for routine use. PMID- 12760998 TI - The effect of hypothermia on myogenic motor-evoked potentials to electrical stimulation with a single pulse and a train of pulses under propofol/ketamine/fentanyl anesthesia in rabbits. AB - In the present study, we investigated the effect of hypothermia on myogenic motor evoked potentials (MEPs) in rabbits. The influence of stimulation paradigms to induce MEPs was evaluated. Twelve rabbits anesthetized with ketamine, fentanyl, and propofol were used for the study. Myogenic MEPs in response to electrical stimulation of the motor cortex with a single pulse and a train of three and five pulses were recorded from the soleus muscle. After the control recording of MEPs at 38 degrees C of esophageal temperature, the rabbits were cooled by surface cooling. Esophageal temperature was maintained at 35 degrees C, 32 degrees C, 30 degrees C, and 28 degrees C, and MEPs were recorded at each point. MEP amplitude to single- pulse stimulation was significantly reduced with a re-duction of core temperature to 28 degrees C compared with the control value at 38 degrees C (0.8 +/- 0.4 mV versus 2.3 +/- 0.3 mV; P < 0.05), whereas MEP amplitude to train-pulse stimulation did not change significantly during the cooling. MEP latency was increased linearly with a reduction of core temperature regardless of stimulation paradigms. In conclusion, these results indicate that a reduction of core temperature to 28 degrees C did not influence MEP amplitudes as long as a train of pulses, but not a single pulse, was used for stimulation in rabbits under propofol/ketamine/fentanyl anesthesia. IMPLICATIONS: Intraoperative monitoring of myogenic motor-evoked potentials (MEPs) may be required under hypothermic conditions because of its neuroprotective efficacy. However, data on the influence of hypothermia on myogenic MEPs are limited. The results indicate that multipulse stimulation may be better than single-pulse stimulation when monitoring MEPs during hypothermia. PMID- 12760999 TI - Interference of electromagnetic operating systems in otorhinolaryngology surgery with bispectral index monitoring. AB - IMPLICATIONS: Electromagnetic operating systems interfere with bispectral index monitoring in creating incorrectly large values; clinicians have to keep this in mind whenever these systems are used. PMID- 12761000 TI - The effect of systemic zonisamide (Zonegran) on thermal hyperalgesia and mechanical allodynia in rats with an experimental mononeuropathy. AB - We studied the ability of zonisamide (Zonegran) to relieve thermal hyperalgesia and/or mechanical allodynia in the chronic constriction injury model of neuropathic pain. Zonisamide (25, 50, or 100 mg/kg) or saline was administered in a blinded, randomized manner by intraperitoneal injection on postoperative days (PODs) 4, 5, and 6. Paw withdrawal latency (PWL) to heat, paw withdrawal response to von Frey monofilaments, and pain scores based on weight-bearing were tested: before surgery; before and after zonisamide or saline (PODs 4, 5, and 6); and on POD 9. Systemic zonisamide relieved thermal hyperalgesia in a dose-dependent manner. All PWLs were significantly increased after zonisamide administration compared with pre-zonisamide measurements, except with the 100 mg/kg dose on POD 5. After zonisamide 100 mg/kg administration, there was a sustained increase in PWL on PODs 5 and 9, with significant carryover effect from the previous dose. However, zonisamide had little effect on mechanical allodynia, except at the 100 mg/kg dose, which was sedating in the rat. At the 100 mg/kg dose, paw withdrawal response was increased on PODs 4 and 5, whereas pain scores were reduced on PODs 4, 5, and 6. Pain scores were inconsistently reduced after 50 mg/kg or 25 mg/kg doses. IMPLICATIONS: Zonisamide causes a dose-related decrease in heat sensitivity in a rat model of neuropathic pain, but relieves mechanical sensitivity only in a dose that is sedating to the rat. Zonisamide may be useful in the treatment of some types of neuropathic pain. PMID- 12761001 TI - Cutaneous analgesia after transdermal application of amitriptyline versus lidocaine in rats. AB - Amitriptyline, a tricyclic antidepressant, has potent local anesthetic properties. However, there is no report of cutaneous analgesic effects after transdermal application. We report here that transdermally applied amitriptyline is more potent than lidocaine in providing cutaneous analgesia in rats. Solutions of amitriptyline base in 50, 100, and 500 mM concentrations were applied as a patch to rats, and their effects were compared with those of lidocaine base at the same concentrations and of the vehicle alone (45% water, 45% isopropyl alcohol, and 10% glycerin). Rats in each test group developed a concentration dependent cutaneous analgesic block in the areas to which the drugs were applied; however, amitriptyline produced a longer block than lidocaine at the same concentration. The development of amitriptyline as a longer-lasting topical analgesic may improve our ability to treat chronic pain, such as neuropathic pain and neuralgia, and to prevent pain in procedures such as venipuncture. IMPLICATIONS: The tricyclic antidepressant amitriptyline, often used perorally for the management of chronic pain, is shown here to be more potent than lidocaine in providing cutaneous analgesia when applied transdermally with an occlusive dressing in rats. PMID- 12761002 TI - Antinociceptive potentiation and attenuation of tolerance by intrathecal electric stimulation in rats. AB - We tested whether intrathecal electric stimulation would reduce the tolerance to chronic morphine use and the severity of precipitated morphine withdrawal. Rats received intrathecal electrode catheter implantation and a continuous intrathecal infusion of morphine (2 nmol/h) or saline for 7 days. Intrathecal electric stimulations (0, 20, or 200 V) were performed once daily during the same period. Daily tail-flick and intrathecal morphine challenge tests were performed to assess the effect of intrathecal electric stimulation on antinociception and tolerance to morphine. Naloxone withdrawal (2 mg/kg) was performed to assess morphine dependence, and changes in spinal neurotransmitters were monitored by microdialysis. The antinociceptive effect of intrathecal morphine was increased by 200 V of electric stimulation. The magnitude of tolerance was decreased in the rats receiving the 2 nmol/h infusion with 200 V of intrathecal electric stimulation compared with the control group (morphine 2 nmol/h alone) (AD(50), 13.6 vs 124.7 nmol). The severity of naloxone-induced withdrawal was less in the rats receiving 200 V of stimulation. Intrathecal stimulation thus enhances analgesia and attenuates naloxone-induced withdrawal symptoms in rats receiving chronic intrathecal morphine infusion. Increases in spinal glycine release may be the underlying mechanism. This method may merit further investigation in the context of the long-term use of intrathecal opioids for controlling chronic pain. IMPLICATIONS: Control of chronic pain is a major health problem. We show here that direct electrical stimulation of the spinal cord in rats enhances analgesia and attenuates naloxone-induced withdrawal symptoms. This may warrant further investigation in the context of long-term use of intrathecal opioids for controlling chronic pain. PMID- 12761003 TI - A retained Racz catheter fragment after epidural neurolysis: implications during magnetic resonance imaging. AB - IMPLICATIONS: A retained ferromagnetic catheter used for epidurolysis obscured diagnostic magnetic resonance imaging of the lumbar spine. The implications of this are discussed in light of other reports of retained catheter fragments obtained from the Food and Drug Administration Manufacturer and Facility Device Experience Database (http://www.fda.gov/cdrh/maude.html). PMID- 12761004 TI - Cyclooxygenase-2 inhibitors. PMID- 12761005 TI - The use of advanced simulation in the training of anesthesiologists to treat chemical warfare casualties. AB - Training anesthesiologists to treat nerve gas intoxication in a mass casualty scenario is a complicated task. The scenario is an unfamiliar medical situation involving the need to decontaminate patients before providing definitive medical treatment, and the need for physical protection to the medical team before decontamination. We describe the development of a simulation-based training program. In one site of a virtual hospital, anesthesiologists were trained in initial airway and breathing resuscitation before decontamination while wearing full protective gear. In another site, they were trained in the treatment of critically-ill patients with combined conventional and chemical injuries or severe intoxication. Intubation simulators of newborn, pediatric, and adult patients, advanced full-scale simulators, and actors simulating patients were used. Initial airway, breathing, and antidotal treatment were performed successfully, with or without full protective gear. The gas mask did not interfere with orotracheal intubation, but limited effective communication within the medical team. Chemical protective gloves were the limiting factor in the performance of medical tasks such as fixing the orotracheal tube. Twenty-two participants (88%) pointed out that the simulated cases represented realistic problems in this scenario, and all 25 participants found the simulated-based training superior to previous traditional training they had in this field. Using advanced simulation, we were able to train anesthesiologists to treat nerve gas intoxication casualties and to learn about the limitations of providing medical care in this setting. IMPLICATIONS: Advanced medical simulation can be used to train anesthesiologists to treat nonconventional warfare casualties. The limitations of medical performance in full protective gear can be learned from this training. PMID- 12761006 TI - Survival with full neurologic recovery after prolonged cardiopulmonary resuscitation with a combination of vasopressin and epinephrine in pigs. AB - We sought to determine the effects of a combination of vasopressin and epinephrine on neurologic recovery in comparison with epinephrine alone and saline placebo alone in an established porcine model of prolonged cardiopulmonary resuscitation (CPR). After 4 min of cardiac arrest, followed by 3 min of basic life support CPR, 17 animals were randomly assigned to receive, every 5 min, either a combination of vasopressin and epinephrine (vasopressin [IU/kg]/epinephrine [ micro g/kg]: 0.4/45, 0.4/45, and 0.8/45; n = 6), epinephrine alone (45, 45, and 200 micro g/kg; n = 6), or saline placebo alone (n = 5). After 22 min of cardiac arrest, including 18 min of CPR, defibrillation was attempted to achieve the return of spontaneous circulation. Aortic diastolic pressure was significantly (P < 0.01) increased 90 s after each of 3 vasopressin/epinephrine injections versus epinephrine alone versus saline placebo alone (mean +/- SEM: 69 +/- 3 mm Hg versus 45 +/- 3 mm Hg versus 29 +/- 2 mm Hg, 63 +/- 4 mm Hg versus 27 +/- 3 mm Hg versus 23 +/- 1 mm Hg, and 52 +/- 4 mm Hg versus 21 +/- 3 mm Hg versus 16 +/- 3 mm Hg, respectively). Spontaneous circulation was restored in six of six vasopressin/epinephrine pigs, whereas six of six epinephrine and five of five saline placebo pigs died (P < 0.01). Neurologic evaluation 24 h after successful resuscitation revealed only an unsteady gait and was normal 5 days after the experiment in all vasopressin/epinephrine-treated animals. In conclusion, in this porcine model of prolonged CPR, repeated vasopressin/epinephrine administration, but not epinephrine or saline placebo alone, ensured long-term survival with full neurologic recovery. IMPLICATIONS: We present a study to evaluate the effects of a combination of vasopressin and epinephrine during prolonged cardiopulmonary resuscitation on neurological outcome in pigs. We found that all pigs treated with a combination of vasopressin and epinephrine could be resuscitated and had full neurologic recovery observed over an entire period of 5 days. PMID- 12761007 TI - An evaluation of the Laryngeal Tube during general anesthesia using mechanical ventilation. AB - The Laryngeal Tube is a new supraglottic ventilatory device for airway management. It has been developed to secure a patent airway during either spontaneous or mechanical ventilation. In this study, we sought to determine the effectiveness of the Laryngeal Tube for primary airway management during routine surgery with mechanical ventilation. One-hundred-seventy-five subjects classified as ASA physical status I and II, scheduled for elective surgery, were included in the study. After the induction of general anesthesia and insertion of a Size 4 Laryngeal Tube, measurements of oxygen saturation, end-tidal CO(2) and isoflurane concentration, and breath-by-breath spirometry data were obtained every 5 min throughout surgery. The lungs were ventilated with volume-controlled mechanical ventilation. The number of attempts taken to insert the Laryngeal Tube and the insertion time were recorded. In 96.6% of patients, it was possible to maintain oxygenation, ventilation, and respiratory mechanics by using mechanical ventilation throughout the surgical procedure. The results of this study suggest that the Laryngeal Tube is an effective and safe airway device for airway management in mechanically ventilated patients during elective surgery. IMPLICATIONS: In 96.6% of patients intubated with the Laryngeal Tube, it was possible to maintain oxygenation, ventilation, and respiratory mechanics during mechanical ventilation. PMID- 12761008 TI - Prone position improves lung mechanical behavior and enhances gas exchange efficiency in mechanically ventilated chronic obstructive pulmonary disease patients. AB - Pronation might favorably affect respiratory system (rs) mechanics and function in volume-controlled, mode-ventilated chronic obstructive pulmonary disease (COPD) patients. We studied 10 COPD patients, initially positioned supine (baseline supine [supine(BAS)]) and then randomly and consecutively changed to protocol supine (supine(PROT)), semirecumbent, and prone positions. Rs mechanics and inspiratory work (W(I)) were assessed at baseline (0.6 L) (all postures) and sigh (1.2 L) (supine(BAS) excluded) tidal volume (V(T)) with rapid airway occlusion during constant-flow inflation. Hemodynamics and gas exchange were assessed in all postures. There were no complications. Prone positioning resulted in (a) increased dynamic-static chest wall (cw) elastance (at both V(Ts)) and improved oxygenation versus supine(BAS), supine(PROT), and semirecumbent, (b) decreased additional lung (L) resistance-elastance versus supine(PROT) and semirecumbent at sigh V(T), (c) decreased L-static elastance (at both V(Ts)) and improved CO(2) elimination versus supine(BAS) and supine(PROT), and (d) improved oxygenation versus all other postures. Semirecumbent positioning increased mainly additional cw-resistance versus supine(BAS) and supine(PROT) at baseline. V(T) W(I)-sub-component changes were consistent with changes in rs, cw, and L mechanical properties. Total rs-W(I) and hemodynamics were unaffected by posture change. After pronation, five patients were repositioned supine (supine(POSTPRO)). In supine(POSTPRO), static rs-L elastance were lower, and oxygenation was still improved versus supine(BAS). Pronation of mechanically ventilated COPD patients exhibits applicability and effectiveness and improves oxygenation and sigh-L mechanics versus semirecumbent ("gold standard") positioning. IMPLICATIONS: By assessing respiratory mechanics, inspiratory work, hemodynamics, and gas exchange, we showed that prone positioning of mechanically ventilated chronic obstructed pulmonary disease patients improves oxygenation and lung mechanics during sigh versus semirecumbent positioning. Furthermore, certain pronation-related benefits versus preprone-supine positioning (reduced lung elastance and improved oxygenation) are maintained in the postprone supine position. PMID- 12761009 TI - Presentation of non-Hodgkin's lymphoma as acute hypoxia caused by right ventricular compression. AB - IMPLICATIONS: A case is presented of a woman developing acute right ventricular outflow tract obstruction because of mediastinal non-Hodgkin's lymphoma. Systemic blood flow was possible through a patent foramen ovale. PMID- 12761010 TI - The effect of graded hypothermia (36 degrees C-32 degrees C) on hemostasis in anesthetized patients without surgical trauma. AB - The isolated effects of hypothermia on hemostasis have not been investigated in healthy humans. We cooled 16 anesthetized patients scheduled for elective intracranial surgery to 32 degrees C body core temperature and assessed prothrombin time (PT), activated partial thromboplastin time, thrombelastogram (TEG), closure time, and platelet count at 36 degrees C, 34 degrees C, and 32 degrees C body core temperature after the induction of anesthesia but before surgical intervention. Activated partial thromboplastin time, hematocrit, and closure time did not change, whereas PT and platelet count decreased during cooling. Platelet count decreased without a decrease in hematocrit; hence, a dilution by administered fluids seemed unlikely. The small decrease of platelet count is probably clinically irrelevant in patients with normal platelet count and function. The small decrease in PT indicates an alteration of the extrinsic pathway of coagulation. TEG measurements showed a delay of clot formation in temperature-adjusted measurements but showed no change if the test temperature was 37 degrees C. This indicates that hypothermia reduces plasmatic coagulation and platelet reactivity. However, the clot strength is not altered by hypothermia. All coagulation variables remained within the normal ranges. Our results may indicate that moderate short-term (4-h) hypothermia has only minor adverse effects in healthy humans. We can make no statement about the effects of hypothermia of longer duration. IMPLICATIONS: This study investigated the isolated effects of hypothermia in healthy anesthetized humans. We found only minor effects of body temperature reduction to 32 degrees C on assessed coagulation variables, indicating only minor effects in otherwise healthy humans. PMID- 12761011 TI - The temporal profile of the reaction of microglia, astrocytes, and macrophages in the delayed onset paraplegia after transient spinal cord ischemia in rabbits. AB - In the present study, we sought to elucidate the temporal profile of the reaction of microglia, astrocytes, and macrophages in the progression of delayed onset motor dysfunction after spinal cord ischemia (15 min) in rabbits. At 2, 4, 8, 12, 24, and 48 h after reperfusion (9 animals in each), hind limb motor function was assessed, and the lumbar spinal cord was histologically examined. Delayed motor dysfunction was observed in most animals at 48 h after ischemia, which could be predicted by a poor recovery of segmental spinal cord evoked potentials at 15 min of reperfusion. In the gray matter of the lumbar spinal cord, both microglia and astrocytes were activated early (2 h) after reperfusion. Microglia were diffusely activated and engulfed motor neurons irrespective of the recovery of segmental spinal cord evoked potentials. In contrast, early astrocytic activation was confined to the area where neurons started to show degeneration. Macrophages were first detected at 8 h after reperfusion and mainly surrounded the infarction area later. Although the precise roles of the activation of microglia, astrocytes, and macrophages are to be further determined, the results indicate that understanding functional changes of astrocytes may be important in the mechanism of delayed onset motor dysfunction including paraplegia. IMPLICATIONS: Microglia and macrophages play a role in removing tissue debris after transient spinal cord ischemia. Disturbance of astrocytic defense mechanism, breakdown of the blood spinal cord barrier, or both seemed to be involved in the development of delayed motor dysfunction. PMID- 12761012 TI - Transient paraplegia revealed by intraoperative neurophysiological monitoring: was it caused by the epidural anesthetic or an epidural hematoma? AB - IMPLICATIONS: Our case report describes the electrophysiological features associated with the development of a spinal epidural hematoma during surgery of the lumbar spine. It stresses the importance of the evaluation of nonsurgical factors, which can potentially affect intraoperative evoked potentials; in this case, epidural local anesthetic or epidural hematoma. PMID- 12761013 TI - Nalbuphine versus ondansetron for prevention of intrathecal morphine-induced pruritus after cesarean delivery. AB - In this prospective, randomized, double-blinded study, we compared the prophylactic efficacy of nalbuphine and ondansetron for the prevention of intrathecal morphine-induced pruritus after cesarean delivery. Two-hundred-forty parturients were randomly allocated into four groups. The N-4 group, O-4 group, O 8 group, and placebo group received IV 4 mg of nalbuphine, 4 mg of ondansetron, 8 mg of ondansetron, and 4 mL of normal saline, respectively, immediately after the baby was delivered. In the postanesthesia care unit, we found that the severity of pruritus score in the four groups was significantly different (P < 0.001). The prophylactic success rate for pruritus of the N-4, O-4, O-8, and placebo groups was 20%, 13%, 12%, and 6%, respectively (P < 0.001). The pruritus score between N 4 and placebo and O-4 and placebo was significantly different (P < 0.001 and P = 0.006, respectively). Treatment for pruritus was requested by patients in 25%, 47%, 51%, and 72% of patients in the N-4, O-4, O-8, and placebo groups, respectively (P < 0.001). There were no differences among groups in nausea/vomiting score, pain score, sedation score, or shivering score at 4, 8, and 24 h after surgery. Nalbuphine and ondansetron are more effective than placebo for the prevention of intrathecal morphine-induced pruritus after cesarean delivery. IMPLICATIONS: Nalbuphine and ondansetron are more effective than placebo for the prevention of intrathecal morphine-induced pruritus after cesarean delivery. PMID- 12761014 TI - The numeric rating scale and labor epidural analgesia. AB - A verbal numeric 0-10 rating scale (NRS) is widely used to evaluate pain in research studies, but its usefulness to the clinician is not well established. In this study, we define desire for additional analgesic medication as a clinically relevant outcome in research studies about pain and compare it with the results of the NRS. A post hoc analysis of three studies that we previously conducted concerning labor epidural analgesia was performed. In all three studies, a verbal NRS score was obtained before and 15 min after labor epidural analgesia. At 15 min, the woman was also asked if she wanted more pain medication. We found that very few patients (2%) with a NRS score of 0-1 wanted more medication. When the NRS score was 2 or 3, 51% of the patients wanted more medication, and when the NRS score was >3, almost all patients (93%) wanted more medication. Grouping the final NRS scores into 3 categories (0 or 1, 2 or 3, and >3) is more useful to the clinician than using individual NRS scores. IMPLICATIONS: This study demonstrated that unless the score of the verbal numeric 0-10 rating scale (NRS) is 0 or 1, most women want more analgesic medication for labor epidural analgesia. Additionally, we found that grouping the NRS values into 3 categories for analysis (0 or 1, 2 or 3, and > 3) is more useful to the clinician than using the full spectrum of NRS scores. PMID- 12761015 TI - Diluent volume for epidural fentanyl and its effect on analgesia in early labor. AB - Epidural fentanyl after a lidocaine and epinephrine test dose provides adequate analgesia and allows for ambulation during early labor. We designed the current study to determine the influence of the diluent volume of the epidural fentanyl bolus (e.g., whether it has an effect on the onset and duration of analgesia). Sixty laboring primigravid women received a 3-mL epidural test dose of lidocaine with epinephrine and then received a fentanyl 100- micro g bolus in either a 2 mL, 10-mL, or 20-mL volume. Pain scores and side effects were recorded for each patient. The onset of analgesia was similar in all three groups. The mean duration before re-dose was not significantly different in the 2-mL group (108 +/ 40 min), the 10-mL group (126 +/- 57 min), or the 20-mL group (126 +/- 41 min). No patient in any group experienced any detectable motor block; one patient (2-mL group) complained of mild knee weakness and was not allowed to ambulate. In early laboring patients, the volume in which 100 micro g of epidural fentanyl (after a lidocaine-epinephrine test dose) is administered does not affect the onset or duration of analgesia, nor does it affect the ability to ambulate. IMPLICATIONS: In early laboring patients, the volume in which 100 micro g of epidural fentanyl (after a lidocaine-epinephrine test dose) is administered does not affect the onset or duration of ambulatory analgesia. PMID- 12761016 TI - Epidural labor analgesia for a patient with disseminated lymphangiomatosis. AB - IMPLICATIONS: We describe a case of a parturient with disseminated lymphangiomatosis involving the thorax, retroperitoneum, and lumbar vertebrae who received epidural labor analgesia. Clinical presentations vary depending on the organ systems involved, the extent of the disease, and the stage of pregnancy. Anesthetic implications are discussed. PMID- 12761017 TI - Postdural puncture headache: an imaging-guided management protocol. AB - IMPLICATIONS: We propose an imaging-based algorithm for the management of headache caused by the inadvertent puncture of dura that occurs sporadically during epidural analgesia. Its implementation can identify those postdural puncture headache cases that cannot benefit from epidural blood patches, and their unnecessary application can consequently be avoided. PMID- 12761018 TI - Does lead interfere with hemoglobin-based oxygen carrier (HBOC) function? A pilot study of lead concentrations in three approved or tested HBOCs and oxyhemoglobin dissociation with HBOCs and/or bovine blood with varying lead concentrations. AB - We measured lead concentrations in three hemoglobin-based oxygen carriers (HBOCs; Oxyglobin, Hemopure, and Hemolink) and compared them with lead concentrations from blood-bank blood. Oxyhemoglobin dissociation was measured with large concentrations of lead in bovine HBOC, with or without bovine blood, and in bovine blood. Samples of each were prepared by combining one with normal saline (control), the second with small lead concentrations (22 micro g/dL), and the third with toxic lead concentrations (70 micro g/dL). They were blended in 2 tonometers at oxygen concentrations (2.5%, 5%, 8%, 10%, 21%, and 95%) with 5% CO(2) and the remainder nitrogen for 5 min per sample after a 15-min wash-in with each level of oxygen and were measured with co-oximetry. Oxygen saturation was plotted against PO(2), fitting fourth-order polynomial nonlinear regression to the data. The lead concentrations of the three HBOCs were 0.51, 0.22, 0.40 micro g/dL. There were no clinically important differences of the oxyhemoglobin dissociation curves as a function of lead concentration. The lead concentrations of the three tested HBOCs were small and no larger than the average for blood bank blood. The presence of increasing concentrations of lead in either concentrated solution of bovine HBOC or a 1:1 mixture of bovine HBOC and native bovine blood does not appear to affect hemoglobin oxygenation in an acute in vitro model of increased lead concentrations. IMPLICATIONS: Gunshot wounds rapidly increase circulating lead concentrations. Lead concentrations are small in three hemoglobin-based oxygen carriers (HBOCs), and HBOCs and/or bovine blood do not appear to be affected by lead concentrations in terms of immediate oxygen on-loading and off-loading. HBOCs may be useful in patients with gunshot wounds. PMID- 12761019 TI - Storage capacities of the laryngeal mask and laryngeal tube compared and their relevance to aspiration risk during positive pressure ventilation. AB - IMPLICATIONS: Supraglottic airways used for positive pressure ventilation run the occasional risk of regurgitated liquid entering the lungs (aspiration). A dynamic model described here shows that the laryngeal tube has a larger liquid storage capacity between the two cuffs than the bowl of the laryngeal mask, with a consequent smaller aspiration risk. PMID- 12761020 TI - Priorities in perioperative geriatrics. PMID- 12761021 TI - A teaching model for training in regional anesthesia--or in peripheral nerve blockade? PMID- 12761022 TI - Are pharmaceutical representatives reliable guides to the medical literature? PMID- 12761023 TI - Another cause of pseudohypotension. PMID- 12761024 TI - Magnesium sulfate and neuroprotection. PMID- 12761025 TI - Pitfalls of indocyanine green dye elimination to assess graft function during liver transplantation. PMID- 12761026 TI - Strategies to prevent arterial injury caused by dilator should be integrated into routine clinical practice. PMID- 12761028 TI - Anesthetic considerations for bariatric surgery: proper positioning is important for laryngoscopy. PMID- 12761027 TI - Combination spinal analgesic chemotherapy: an additional clinical trial of opioid plus local anesthetic. PMID- 12761029 TI - Anesthesia of a patient with cured myasthenia gravis. PMID- 12761030 TI - A new laryngeal tube. PMID- 12761031 TI - Preoperative risk factors of intraoperative hypothermia in major surgery under general anesthesia. PMID- 12761032 TI - Was case report a case of unrecognized local anesthetic toxicity? PMID- 12761033 TI - Certification in perioperative TEE. PMID- 12761034 TI - Use of gum elastic bougie during difficult airway management. PMID- 12761035 TI - The impact of a cardioprotective protocol on the incidence of cardiac complications after aortic abdominal surgery. PMID- 12761036 TI - Upregulation of the Rac1/JNK signaling pathway in primary human schwannoma cells. AB - Schwann cells lacking the tumor-suppressor-protein merlin tend in man to build benign tumors (schwannoma). We observed that characteristic features of these cells which are relevant to tumorigenicity resemble those described in cells with high Rac activity. Moreover this small GTPase also phosphorylates merlin via PAK activation. We hypothesized that merlin deficiency might cause an activation of Rac and its dependent signaling pathways, in particular the pro-tumorigenic JNK pathway. We show an enhanced activation of Rac1 in primary human schwannoma cells, find both Rac and its effector PAK at the membrane where they colocalize, and describe increased levels of phosphorylated JNK in the nucleus of these cells. Further we describe regulation at post-transcriptional level with upregulated protein, but not mRNA levels for Rac1, and JNK1/2. We conclude that merlin regulates Rac activation, and suggest that this is important for human schwannoma cell dedifferentiation. PMID- 12761037 TI - Identification and functional characterization of a novel R621C mutation in the synphilin-1 gene in Parkinson's disease. AB - Synphilin-1 is linked to the pathogenesis of Parkinson's disease (PD) based on its identification as an alpha-synuclein (PARK1) and parkin (PARK2) interacting protein. Moreover, synphilin-1 is a component of Lewy bodies (LB) in brains of sporadic PD patients. Therefore, we performed a detailed mutation analysis of the synphilin-1 gene in 328 German familial and sporadic PD patients. In two apparently sporadic PD patients we deciphered a novel C to T transition in position 1861 of the coding sequence leading to an amino acid substitution from arginine to cysteine in position 621 (R621C). This mutation was absent in a total of 702 chromosomes of healthy German controls. To define a possible role of mutant synphilin-1 in the pathogenesis of PD we performed functional analyses in SH-SY5Y cells. We found synphilin-1 capable of producing cytoplasmic inclusions in transfected cells. Moreover we observed a significantly reduced number of inclusions in cells expressing C621 synphilin-1 compared with cells expressing wild-type (wt) synphilin-1, when subjected to proteasomal inhibition. C621 synphilin-1 transfected cells were more susceptible to staurosporine-induced cell death than cells expressing wt synphilin-1. Our findings argue in favour of a causative role of the R621C mutation in the synphilin-1 gene in PD and suggest that the formation of intracellular inclusions may be beneficial to cells and that a mutation in synphilin-1 that reduces this ability may sensitize neurons to cellular stress. PMID- 12761038 TI - Therapeutic benefits of cardiotrophin-1 gene transfer in a mouse model of spinal muscular atrophy. AB - Spinal muscular atrophy (SMA) is a recessive autosomal disorder characterized by degeneration of lower motor neurons caused by mutations of the survival motor neuron gene (SMN1). No curative treatment is known so far. Mutant mice carrying homozygous deletion of Smn exon 7 directed to neurons display skeletal muscle denervation, moderate loss of motor neuron cell bodies and severe axonal degeneration. These features, similar to those found in human SMA, strongly suggest the involvement of a dying back process of motor neurons and led us to test whether neurotrophic factors might have a protective role in SMA. We report here the therapeutic benefits of systemic delivery of cardiotrophin-1 (CT-1), a neurotrophic factor belonging to the IL-6 cytokine family. Intra-muscular injection of adenoviral vector expressing CT-1, even at very low dose, improves median survival, delays motor defect of mutant mice and exerts protective effect against loss of proximal motor axons and aberrant cytoskeletal organization of motor synaptic terminals. In spite of the severity of SMA phenotype in mutant mice, CT-1 is able to slow down disease progression. Neuroprotection could be regarded as valuable therapeutic approach in SMA. PMID- 12761039 TI - Genetic and functional analyses of FH mutations in multiple cutaneous and uterine leiomyomatosis, hereditary leiomyomatosis and renal cancer, and fumarate hydratase deficiency. AB - Germline mutations of the fumarate hydratase (FH, fumarase) gene are found in the recessive FH deficiency syndrome and in dominantly inherited susceptibility to multiple cutaneous and uterine leiomyomatosis (MCUL). We have previously reported a number of germline FH mutations from MCUL patients. In this study, we report additional FH mutations in MCUL and FH deficiency patients. Mutations can readily be found in about 75% of MCUL cases and most cases of FH deficiency. Some of the more common FH mutations are probably derived from founding individuals. Protein truncating FH mutations are functionally null alleles. Disease-associated missense FH changes map to highly conserved residues, mostly in or around the enzyme's active site or activation site; we predict that these mutations severely compromise enzyme function. The mutation spectra in FH deficiency and MCUL are similar, although in the latter mutations tend to occur earlier in the gene and, perhaps, are more likely to result in a truncated or absent protein. We have found that not all mutation-carrier parents of FH deficiency children have a strong predisposition to leiomyomata. We have confirmed that renal carcinoma is sometimes part of MCUL, as part of the variant hereditary leiomyomatosis and renal cancer (HLRCC) syndrome, and have shown that these cancers may have either type II papillary or collecting duct morphology. We have found no association between the type or site of FH mutation and any aspect of the MCUL phenotype. Biochemical assay for reduced FH functional activity in the germline of MCUL patients can indicate carriers of FH mutations with high sensitivity and specificity, and can detect reduced FH activity in some patients without detectable FH mutations. We conclude that MCUL is probably a genetically homogeneous tumour predisposition syndrome, primarily resulting from absent or severely reduced fumarase activity, with currently unknown functional consequences for the smooth muscle or kidney cell. PMID- 12761040 TI - Prevention of polyglutamine oligomerization and neurodegeneration by the peptide inhibitor QBP1 in Drosophila. AB - Polyglutamine (polyQ) diseases are a growing class of inherited neurodegenerative diseases including Huntington's disease, which are caused by abnormal expansions of the polyQ stretch in each unrelated disease protein. The expanded polyQ stretch is thought to confer toxic properties on the disease proteins through alteration of their conformation leading to pathogenic protein-protein interactions including oligomerization and/or aggregation. Hypothesizing that molecules with selective binding affinity to the expanded polyQ stretch may interfere with the pathogenic properties, we previously identified Polyglutamine Binding Peptide 1 (QBP1) from combinatorial peptide phage display libraries. We show here that a tandem repeat of the inhibitor peptide QBP1, (QBP1)(2), significantly suppresses polyQ aggregation and polyQ-induced neurodegeneration in the compound eye of Drosophila polyQ disease models, which express the expanded polyQ protein under the eye specific promoter. Most importantly, (QBP1)(2) expression dramatically rescues premature death of flies expressing the expanded polyQ protein in the nervous system, resulting in the dramatic increase of the median life span from 5.5 to 52 days. These results suggest that QBP1 can prevent polyQ-induced neurodegeneration in vivo. We propose that QBP1 prevents polyQ oligomerization and/or aggregation either by altering the toxic conformation of the expanded polyQ stretch, or by simply competing with the expanded polyQ stretches for binding to other expanded polyQ proteins. The peptide inhibitor QBP1 is a promising candidate with great potential as a therapeutic molecule against the currently untreatable polyQ diseases. PMID- 12761041 TI - Gene expression differences in quiescent versus regenerating hair cells of avian sensory epithelia: implications for human hearing and balance disorders. AB - The sensory receptors for hearing and balance are the hair cells of the cochlea and vestibular organs of the inner ear. Permanent hearing and balance deficits can be triggered by genetic susceptibilities or environmental factors such as infection. Unlike mammalian hair cells that have a limited capacity for regeneration, the vestibular organ of the avian ear is constantly undergoing hair cell regeneration, whereas the avian cochlea undergoes regeneration only when hair cells are damaged. In order to gain insights into the genetic programs that govern the regenerative capacity of hair cells, we interrogated custom human cDNA microarrays with sensory epithelial cell targets from avian inner ears. The arrays contained probes from conserved regions of approximately 400 genes expressed primarily in the inner ear and approximately 1500 transcription factors (TF). Highly significant differences were observed for 20 inner-ear genes and more than 80 TFs. Genes up-regulated in the cochlea included BMP4, GATA3, GSN, FOXF1 and PRDM7. Genes up-regulated in the utricle included SMAD2, KIT, beta AMYLOID, LOC51637, HMG20B and CRIP2. Many of the highly significant changes were validated by Q-PCR and in situ methods. Some of the observed changes implicated a number of known biochemical pathways including the c-kit pathway previously observed in melanogenesis. Twenty differentially expressed TFs map to chromosomal regions harboring uncloned human deafness loci, and represent novel candidates for hearing loss. The approach described here also illustrates the power of utilizing conserved human cDNA probes for cross-species comparisons. PMID- 12761042 TI - Genome-wide linkage reveals a locus for human essential (primary) hypertension on chromosome 12p. AB - Essential (primary) hypertension is an important risk factor for cardiovascular morbidity and mortality. Blood pressure is largely heritable; however, the genetic factors contributing to essential hypertension are mostly unknown. We examined a large Chinese kindred (n=387) and selected a subset of 94 individuals for genotyping. An additional 32 Chinese nuclear families with essential hypertension were also recruited. Genome-wide parametric linkage analysis identified a new locus for primary hypertension on chromosome 12p (parametric LOD score 3.44). This locus overlaps with the assigned locus that causes severe autosomal-dominant hypertension and brachydactyly, the only form of monogenic hypertension known to date that resembles primary hypertension. We suggest that this genomic region, spanning 18 annotated genes, will be of great relevance in elucidating new mechanisms for primary hypertension. PMID- 12761043 TI - Association of Eotaxin gene family with asthma and serum total IgE. AB - The Eotaxin gene family (Eotaxin1, Eotaxin2 and Eotaxin3) recruits and activates CCR3-bearing cells such as eosinophils, mast cells and Th2 lymphocytes that play a major role in allergic disorders. To date, the effect of polymorphisms of Eotaxin genes on asthma phenotypes has not been thoroughly examined. In our research, we sequenced whole regions of the Eotaxin gene family to identify polymorphisms, which may be involved in the development of asthma and total serum IgE. We have identified 37 SNPs in the Exotaxin gene family (Exotaxin1, 2 and 3), and 17 common polymorphic sites were selected for genotyping in our asthma cohort (n=721). Statistical analysis revealed that the EOT2+1265A>G G* allele showed significantly lower frequency in asthmatics than in normal healthy controls (0.14 versus 0.23, P=0.002), and that distribution of the EOT2+1265A>G G* allele containing genotypes was also much lower in asthmatics (26.3 versus 40.8%, P=0.003). In addition, a non-synonymous SNP in Eotaxin1, EOT1+123Ala>Thr showed significant association with total serum IgE levels (P=0.002-0.02). The effect of EOT1+123Ala>Thr on total serum IgE appeared in a gene-dose-dependent manner. Our findings suggest that the development of asthma may be associated with EOT2+1265A>G polymorphisms, and the susceptibility to high IgE production may be attributed to the EOT1+123Ala>Thr polymorphism. Eotaxin variation/haplotype information identified in this study might provide valuable insights into strategies for the control of asthma. PMID- 12761044 TI - Prolonged dystrophin expression and functional correction of mdx mouse muscle following gene transfer with a helper-dependent (gutted) adenovirus-encoding murine dystrophin. AB - Dystrophin gene transfer using helper-dependent adenoviruses (HDAd), which are deleted of all viral genes, is a promising option to treat muscles in Duchenne muscular dystrophy. We investigated the benefits of this approach by injecting the tibialis anterior (TA) muscle of neonatal and juvenile (4-6-week-old) dystrophin-deficient (mdx) mice with a fully deleted HDAd (HDCBDysM). This vector encoded two full-length murine dystrophin cDNAs regulated by the powerful cytomegalovirus enhancer/beta-actin promoter. At 10 days post-injection of neonatal muscles, 712 fibers (42% of the total number of TA fibers) were dystrophin-positive (dys+), a value that did not decrease for 6 months (the study duration). In treated juveniles, maximal transduction occurred at 30 days post injection (414 dys+ fibers, 24% of the total number of TA fibers), but decreased by 51% after 6 months. All studied aspects of the pathology were improved in neonatally treated muscles: the percentage of dys+ fibers with centrally localized myonuclei remained low, localization of the dystrophin associated protein complex was restored at the plasma membrane, muscle hypertrophy was reduced, and maximal force-generating capacity and resistance to contraction induced injuries were increased. The same pathological aspects were improved in the treated juveniles, except for reduction of muscle hypertrophy and maximal force-generating capacity. We demonstrated a strong humoral response against murine dystrophin in both animal groups, but mild inflammatory response occurred only in the treated juveniles. HDCBDysM is thus one of the most promising and efficient vectors for treating DMD by gene therapy. PMID- 12761045 TI - An engineered 800 kilobase deletion of Uchl3 and Lmo7 on mouse chromosome 14 causes defects in viability, postnatal growth and degeneration of muscle and retina. AB - The Acrg minimal region is a 1.5-1.7 Mb domain defined by genetic complementation among deletions generated around Ednrb on chromosome 14 in mice. Mice homozygous for one of the deletions, Ednrb(s-1Acrg), exhibit embryonic lethality with defects associated with mesoderm development. We predicted that the region contains a single cluster of four genes that encode a TBC domain-containing protein (KIAA0603), a novel protein AK000009, the ubiquitin C-terminal hydrolase L3 (UCHL3) and an F-box/PDZ/LIM domain protein LMO7. A targeted internal deletion of Uchl3 (Uchl3(Delta3-7)) produced viable mice, eliminating this gene as a candidate for the embryonic lethality. To dissect the Acrg minimal region further, we utilized Cre-loxP-mediated chromosome engineering to generate a targeted 800 kb deletion (Lmo7(Delta800)) that removes the distal portion of the region. The deletion includes Uchl3, Lmo7 and an additional 500 kb downstream of the 3' end of Lmo7 where no genes are thought to reside. We found that approximately 40% of mice homozygous for this deletion die between birth and weaning, and are severely runted. The remaining homozygotes are viable, thus ruling out Lmo7 as a single gene candidate for the Ednrb(s-1Acrg) embryonic lethality. Both Uchl3(Delta3-7) and Lmo7(Delta800) mutants displayed retinal degeneration, muscular degeneration and growth retardation, but the severity of the muscular degeneration and growth retardation were enhanced in Lmo7(Delta800) homozygotes. We suggest that the increase in severity may reflect an interaction between Uchl3 and Lmo7 in the ubiquitin-mediated protein degradation pathway. PMID- 12761046 TI - Low conservation of alternative splicing patterns in the human and mouse genomes. AB - Alternative splicing has recently emerged as a major mechanism of generating protein diversity in higher eukaryotes. We compared alternative splicing isoforms of 166 pairs of orthologous human and mouse genes. As the mRNA and EST libraries of human and mouse are not complete and thus cannot be compared directly, we instead analyzed whether known cassette exons or alternative splicing sites from one genome are conserved in the other genome. We demonstrate that about half of the analyzed genes have species-specific isoforms, and about a quarter of elementary alternatives are not conserved between the human and mouse genomes. The detailed results of this study are available at www.ig-msk.ru:8005/HMG_paper. PMID- 12761047 TI - Trans mobilization of genomic DNA as a mechanism for retrotransposon-mediated exon shuffling. AB - Exon shuffling, the juxtaposition and new combinations of exons from different genes, facilitates evolutionary changes by increasing protein diversity or by generating new function. Exon shuffling is generated as a consequence of segmental duplications. Long interspersed element (LINE)-1 (L1)-mediated 3' transduction is a potential pathway for exon shuffling by which L1 associates 3' flanking DNA in cis as a read-through transcript and carries the DNA to a new genomic location. In this pathway, however, the targets are limited to the regions located 3' to L1s. Here we propose that the genomic DNA distant from L1 may be mobilized by an alternative (trans) action of L1. A partial ATM sequence containing a single exon and flanking introns has been retrotransposed to a new genomic location on chromosome 7. There was no L1 around the exon of the authentic ATM locus. An unusual feature that the poly(A) tail tagged to the transposed sequence oriented oppositely to the ATM's transcriptional orientation suggests that a trans action of reverse transcriptase on antisense transcript has driven the duplication of genomic DNA without removing introns. Taking account of similar duplication events in previous studies, a certain class of segmental duplications in the human genome may be accounted for by the trans action of retrotransposon machinery. PMID- 12761048 TI - Telomere length and the expression of natural telomeric genes in human fibroblasts. AB - Progressive telomere shortening occurs with division of normal human cells, and eventually leads to replicative senescence. The mechanism by which the shortened telomeres cause growth arrest is largely unknown. Transcriptional silencing of genes adjacent to telomeres, also called telomere position effect, has been hypothesized as a possible mechanism of telomere-mediated senescence. However, there is no report regarding telomere position effect on natural telomeric genes in human cells. To address whether the expression of natural telomeric genes is regulated by telomere length, we combined quantitative RT-PCR with quantitative fluorescence in situ hybridization to comparatively analyze the expression of 34 telomeric genes and telomere length of their 24 corresponding chromosome ends in young and senescent human fibroblasts. We have demonstrated here that telomere length alone is not sufficient to determine the expression status of natural telomeric genes. An extended analysis of a tandem of eight telomeric genes on a single chromosome end revealed a discontinuous pattern of changed expression during telomere shortening and some of the changes are senescence-specific rather than non-dividing-related. These results suggest that the expression of natural telomeric genes may be influenced by alteration of local heterochromatin structure. PMID- 12761049 TI - HnRNP G and Tra2beta: opposite effects on splicing matched by antagonism in RNA binding. AB - The hnRNP G family comprises three closely related proteins, hnRNP G, RBMY and hnRNP G-T. We showed previously that they interact with splicing activator proteins, particularly hTra2beta, and suggested that they were involved in regulating Tra2-dependent splicing. We show here that hnRNP G and hTra2beta have opposite effects upon the incorporation of several exons, both being able to act as either an activator or a repressor. HnRNP G acts via a specific sequence to repress the skeletal muscle-specific exon (SK) of human slow skeletal alpha tropomyosin, TPM3, and stimulates inclusion of the alternative non-muscle exon. The binding of hnRNP G to the exon is antagonized by hTra2beta. The two proteins also have opposite effects upon a dystrophin pseudo-exon. This exon is incorporated in a patient to a higher level in heart muscle than skeletal muscle, causing X-linked dilated cardiomyopathy. It is included to a higher level after transfection of a mini-gene into rodent cardiac myoblasts than into skeletal muscle myoblasts. Co-transfection with hnRNP G represses incorporation in cardiac myoblasts, whereas hTra2beta increases it in skeletal myoblasts. Both the cell specificity and the protein responses depend upon exon sequences. Since the ratio of hnRNP G to Tra2beta mRNA in humans is higher in skeletal muscle than in heart muscle, we propose that the hnRNP G/Tra2beta ratio contributes to the cellular splicing preferences and that the higher proportion of hnRNP G in skeletal muscle plays a role in preventing the incorporation of the pseudo-exon and thus in preventing skeletal muscle dystrophy. PMID- 12761050 TI - Nuclear interaction of the dynein light chain LC8a with the TRPS1 transcription factor suppresses the transcriptional repression activity of TRPS1. AB - The TRPS1 gene codes for a 1281 amino acids nuclear transcription factor with an unusual combination of different types of zinc finger motifs, including GATA-type DNA-binding and IKAROS-like zinc fingers. TRPS1 is a repressor of GATA-regulated genes and implicated in the human tricho-rhino-phalangeal syndromes. We found that two distinct regions of TRPS1 can physically interact with the dynein light chain 8 protein, LC8a, that are at least 458 amino acids apart from each other. Region A covers 89 amino acids (635-723), spanning three potential C(2)H(2) zinc finger structures, and region B covers the 100 most C-terminal amino acids (1182 1281) containing the IKAROS-like motif. LC8a is known to interact with more than 10 different molecules, both proteins and nucleic acids. In most cases, LC8a was identified as a transport molecule in the cytoplasm. Interestingly, we found that LC8a co-localizes with TRPS1 in dot-like structures in the cell nucleus. In an electrophoretic mobility shift assay we could show that the interaction of LC8a and TRPS1 lowers the binding of TRPS1 to the GATA consensus sequence. In addition, GATA-regulated reporter gene assay indicated that LC8a is able to suppress the transcriptional repression activity of TRPS1. PMID- 12761051 TI - Inference of transcriptional regulation relationships from gene expression data. AB - MOTIVATION: In order to find gene regulatory networks from microarray data, it is important to first find direct regulatory relationships between pairs of genes. RESULTS: We propose a new method for finding potential regulatory relationships between pairs of genes from microarray time series data and apply it to expression data for cell-cycle related genes in yeast. We compare our algorithm, dubbed the event method, with the earlier correlation method and the edge detection method by Filkov et al. When tested on known transcriptional regulation genes, all three methods are able to find similar numbers of true positives. The results indicate that our algorithm is able to identify true positive pairs that are different from those found by the two other methods. We also compare the correlation and the event methods using synthetic data and find that typically, the event method obtains better results. AVALIABILITY: software is available upon request. PMID- 12761052 TI - FPV: fast protein visualization using Java 3D. AB - MOTIVATION: Many tools have been developed to visualize protein structures. Tools that have been based on Java 3D((TM)) are compatible among different systems and they can be run remotely through web browsers. However, using Java 3D for visualization has some performance issues with it. The primary concerns about molecular visualization tools based on Java 3D are in their being slow in terms of interaction speed and in their inability to load large molecules. This behavior is especially apparent when the number of atoms to be displayed is huge, or when several proteins are to be displayed simultaneously for comparison. RESULTS: In this paper we present techniques for organizing a Java 3D scene graph to tackle these problems. We have developed a protein visualization system based on Java 3D and these techniques. We demonstrate the effectiveness of the proposed method by comparing the visualization component of our system with two other Java 3D based molecular visualization tools. In particular, for van der Waals display mode, with the efficient organization of the scene graph, we could achieve up to eight times improvement in rendering speed and could load molecules three times as large as the previous systems could. AVAILABILITY: EPV is freely available with source code at the following URL: http://www.cs.ucsb.edu/~tcan/fpv/ PMID- 12761053 TI - Integrative approach for computationally inferring protein domain interactions. AB - MOTIVATION: The current need for high-throughput protein interaction detection has resulted in interaction data being generated en masse through such experimental methods as yeast-two-hybrids and protein chips. Such data can be erroneous and they often do not provide adequate functional information for the detected interactions. Therefore, it is useful to develop an in silico approach to further validate and annotate the detected protein interactions. RESULTS: Given that protein-protein interactions involve physical interactions between protein domains, domain-domain interaction information can be useful for validating, annotating, and even predicting protein interactions. However, large scale, experimentally determined domain-domain interaction data do not exist. Here, we describe an integrative approach to computationally derive putative domain interactions from multiple data sources, including protein interactions, protein complexes, and Rosetta Stone sequences. We further prove the usefulness of such an integrative approach by applying the derived domain interactions to predict and validate protein-protein interactions. AVAILABILITY: A database of putative protein domain interactions derived using the method described in this paper is available at http://interdom.lit.org.sg. PMID- 12761054 TI - Pathways database system: an integrated system for biological pathways. AB - MOTIVATION: During the next phase of the Human Genome Project, research will focus on functional studies of attributing functions to genes, their regulatory elements, and other DNA sequences. To facilitate the use of genomic information in such studies, a new modeling perspective is needed to examine and study genome sequences in the context of many kinds of biological information. Pathways are the logical format for modeling and presenting such information in a manner that is familiar to biological researchers. RESULTS: In this paper we present an integrated system, called Pathways Database System, with a set of software tools for modeling, storing, analyzing, visualizing, and querying biological pathways data at different levels of genetic, molecular, biochemical and organismal detail. The novel features of the system include: (a) genomic information integrated with other biological data and presented from a pathway, rather than from the DNA sequence, perspective; (b) design for biologists who are possibly unfamiliar with genomics, but whose research is essential for annotating gene and genome sequences with biological functions; (c) database design, implementation and graphical tools which enable users to visualize pathways data in multiple abstraction levels, and to pose predetermined queries; and (d) an implementation that allows for web(XML)-based dissemination of query outputs (i.e. pathways data) to researchers in the community, giving them control on the use of pathways data. AVAILABILITY: Available on request from the authors. PMID- 12761055 TI - Terminology-driven mining of biomedical literature. AB - MOTIVATION: With an overwhelming amount of textual information in molecular biology and biomedicine, there is a need for effective literature mining techniques that can help biologists to gather and make use of the knowledge encoded in text documents. Although the knowledge is organized around sets of domain-specific terms, few literature mining systems incorporate deep and dynamic terminology processing. RESULTS: In this paper, we present an overview of an integrated framework for terminology-driven mining from biomedical literature. The framework integrates the following components: automatic term recognition, term variation handling, acronym acquisition, automatic discovery of term similarities and term clustering. The term variant recognition is incorporated into terminology recognition process by taking into account orthographical, morphological, syntactic, lexico-semantic and pragmatic term variations. In particular, we address acronyms as a common way of introducing term variants in biomedical papers. Term clustering is based on the automatic discovery of term similarities. We use a hybrid similarity measure, where terms are compared by using both internal and external evidence. The measure combines lexical, syntactical and contextual similarity. Experiments on terminology recognition and clustering performed on a corpus of MEDLINE abstracts recorded the precision of 98 and 71% respectively. AVAILABILITY: software for the terminology management is available upon request. PMID- 12761056 TI - Corrected small-sample estimation of the Bayes error. AB - MOTIVATION: A major problem of pattern classification is estimation of the Bayes error when only small samples are available. One way to estimate the Bayes error is to design a classifier based on some classification rule applied to sample data, estimate the error of the designed classifier, and then use this estimate as an estimate of the Bayes error. Relative to the Bayes error, the expected error of the designed classifier is biased high, and this bias can be severe with small samples. RESULTS: This paper provides a correction for the bias by subtracting a term derived from the representation of the estimation error. It does so for Boolean classifiers, these being defined on binary features. Although the general theory applies to any Boolean classifier, a model is introduced to reduce the number of parameters. A key point is that the expected correction is conservative. Properties of the corrected estimate are studied via simulation. The correction applies to binary predictors because they are mathematically identical to Boolean classifiers. In this context the correction is adapted to the coefficient of determination, which has been used to measure nonlinear multivariate relations between genes and design genetic regulatory networks. An application using gene-expression data from a microarray experiment is provided on the website http://gspsnap.tamu.edu/smallsample/ (user:'smallsample', password:'smallsample)'). PMID- 12761057 TI - Computation method to identify differential allelic gene expression and novel imprinted genes. AB - MOTIVATION: Genomic imprinting plays an important role in both normal development and diseases. Abnormal imprinting is strongly associated with several human diseases including cancers. Most of the imprinted genes were discovered in the neighborhood of the known imprinted genes. This approach is difficult to extend to analyze the whole genome. We have decided to take a computational approach to systematically search the whole genome for the presence of mono-allelic expressed genes and imprinted genes in human genome. RESULTS: A computational method was developed to identify novel imprinted or mono-allelic genes. Individuals represented in human cDNA libraries were genotyped using Bayesian statistics, and differential expression of polymorphic alleles was identified. A significant reduction in the number of libraries that expressed both alleles, measured by Z statistics, is a strong indicator for an imprinted or a mono-allelic gene. AVAILABILITY: The data sets are available at http://leelab.nci.nih.gov/leelab/jsp/IGDM/IGDM.html PMID- 12761058 TI - Microarray standard data set and figures of merit for comparing data processing methods and experiment designs. AB - MOTIVATION: There is a very large and growing level of effort toward improving the platforms, experiment designs, and data analysis methods for microarray expression profiling. Along with a growing richness in the approaches there is a growing confusion among most scientists as to how to make objective comparisons and choices between them for different applications. There is a need for a standard framework for the microarray community to compare and improve analytical and statistical methods. RESULTS: We report on a microarray data set comprising 204 in-situ synthesized oligonucleotide arrays, each hybridized with two-color cDNA samples derived from 20 different human tissues and cell lines. Design of the approximately 24 000 60mer oligonucleotides that report approximately 2500 known genes on the arrays, and design of the hybridization experiments, were carried out in a way that supports the performance assessment of alternative data processing approaches and of alternative experiment and array designs. We also propose standard figures of merit for success in detecting individual differential expression changes or expression levels, and for detecting similarities and differences in expression patterns across genes and experiments. We expect this data set and the proposed figures of merit will provide a standard framework for much of the microarray community to compare and improve many analytical and statistical methods relevant to microarray data analysis, including image processing, normalization, error modeling, combining of multiple reporters per gene, use of replicate experiments, and sample referencing schemes in measurements based on expression change. AVAILABILITY/SUPPLEMENTARY INFORMATION: Expression data and supplementary information are available at http://www.rii.com/publications/2003/HE_SDS.htm PMID- 12761059 TI - Approximate variance-stabilizing transformations for gene-expression microarray data. AB - MOTIVATION: A variance stabilizing transformation for microarray data was recently introduced independently by several research groups. This transformation has sometimes been called the generalized logarithm or glog transformation. In this paper, we derive several alternative approximate variance stabilizing transformations that may be easier to use in some applications. RESULTS: We demonstrate that the started-log and the log-linear-hybrid transformation families can produce approximate variance stabilizing transformations for microarray data that are nearly as good as the generalized logarithm (glog) transformation. These transformations may be more convenient in some applications. PMID- 12761060 TI - Fuzzy C-means method for clustering microarray data. AB - MOTIVATION: Clustering analysis of data from DNA microarray hybridization studies is essential for identifying biologically relevant groups of genes. Partitional clustering methods such as K-means or self-organizing maps assign each gene to a single cluster. However, these methods do not provide information about the influence of a given gene for the overall shape of clusters. Here we apply a fuzzy partitioning method, Fuzzy C-means (FCM), to attribute cluster membership values to genes. RESULTS: A major problem in applying the FCM method for clustering microarray data is the choice of the fuzziness parameter m. We show that the commonly used value m = 2 is not appropriate for some data sets, and that optimal values for m vary widely from one data set to another. We propose an empirical method, based on the distribution of distances between genes in a given data set, to determine an adequate value for m. By setting threshold levels for the membership values, genes which are tigthly associated to a given cluster can be selected. Using a yeast cell cycle data set as an example, we show that this selection increases the overall biological significance of the genes within the cluster. AVAILABILITY: Supplementary text and Matlab functions are available at http://www-igbmc.u-strasbg.fr/fcm/ PMID- 12761062 TI - Codon pairs in the genome of Escherichia coli. AB - MOTIVATION: The effect of two neighboring codons (codon pairs) on gene expression is mediated via the interaction of their cognate tRNAs occupying the two functional ribosomal sites during the translation elongation step. For steric reasons it is reasonable to assume that not all combinations of codons and therefore of tRNAs are equally favorable when situated on the ribosome surface. Aiming of identifying preferential and rare codon pairs, we have determined the frequency of occurrence of all possible combinations of codon pairs in the entire genome of Escherichia coli (E.coli). RESULTS: The frequency of occurrence of the 3904 codon pairs comprising both sense:sense and sense:stop codon pairs in the full set of E.coli 4289 ORFs was found to vary from zero to 4913 times. For most of the pairs we have observed a significant difference between the real and statistically predicted frequency of occurrence. The analysis of 334 highly expressed and 303 poorly expressed E.coli genes showed that codon pair usage is different for the two gene categories. Using an especially defined criterion (Delta(REG)), the codon pairs are classified as 'hypothetically attenuating' (HAP) and 'hypothetically non-attenuating' (HNAP) and their possible effect on translation is discussed. AVAILABILITY: The program used in this study is available at http://www.bio21.bas.bg/codonpairs/ PMID- 12761061 TI - Mining HIV dynamics using independent component analysis. AB - MOTIVATION: We implement a data mining technique based on the method of Independent Component Analysis (ICA) to generate reliable independent data sets for different HIV therapies. We show that this technique takes advantage of the ICA power to eliminate the noise generated by artificial interaction of HIV system dynamics. Moreover, the incorporation of the actual laboratory data sets into the analysis phase offers a powerful advantage when compared with other mathematical procedures that consider the general behavior of HIV dynamics. RESULTS: The ICA algorithm has been used to generate different patterns of the HIV dynamics under different therapy conditions. The Kohonen Map has been used to eliminate redundant noise in each pattern to produce a reliable data set for the simulation phase. We show that under potent antiretroviral drugs, the value of the CD4+ cells in infected persons decreases gradually by about 11% every 100 days and the levels of the CD8+ cells increase gradually by about 2% every 100 days. AVAILABILITY: Executable code and data libraries are available by contacting the corresponding author. IMPLEMENTATION: Mathematica 4 has been used to simulate the suggested model. A Pentium III or higher platform is recommended. PMID- 12761063 TI - Generating consensus sequences from partial order multiple sequence alignment graphs. AB - MOTIVATION: Consensus sequence generation is important in many kinds of sequence analysis ranging from sequence assembly to profile-based iterative search methods. However, how can a consensus be constructed when its inherent assumption that the aligned sequences form a single linear consensus-is not true? RESULTS: Partial Order Alignment (POA) enables construction and analysis of multiple sequence alignments as directed acyclic graphs containing complex branching structure. Here we present a dynamic programming algorithm (heaviest_bundle) for generating multiple consensus sequences from such complex alignments. The number and relationships of these consensus sequences reveals the degree of structural complexity of the source alignment. This is a powerful and general approach for analyzing and visualizing complex alignment structures, and can be applied to any alignment. We illustrate its value for analyzing expressed sequence alignments to detect alternative splicing, reconstruct full length mRNA isoform sequences from EST fragments, and separate paralog mixtures that can cause incorrect SNP predictions. AVAILABILITY: The heaviest_bundle source code is available at http://www.bioinformatics.ucla.edu/poa PMID- 12761064 TI - ProPred1: prediction of promiscuous MHC Class-I binding sites. AB - SUMMARY: ProPred1 is an on-line web tool for the prediction of peptide binding to MHC class-I alleles. This is a matrix-based method that allows the prediction of MHC binding sites in an antigenic sequence for 47 MHC class-I alleles. The server represents MHC binding regions within an antigenic sequence in user-friendly formats. These formats assist user in the identification of promiscuous MHC binders in an antigen sequence that can bind to large number of alleles. ProPred1 also allows the prediction of the standard proteasome and immunoproteasome cleavage sites in an antigenic sequence. This server allows identification of MHC binders, who have the cleavage site at the C terminus. The simultaneous prediction of MHC binders and proteasome cleavage sites in an antigenic sequence leads to the identification of potential T-cell epitopes. AVAILABILITY: Server is available at http://www.imtech.res.in/raghava/propred1/. Mirror site of this server is available at http://bioinformatics.uams.edu/mirror/propred1/ SUPPLEMENTARY INFORMATION: Matrices and document on server are available at http://www.imtech.res.in/raghava/propred1/page2.html PMID- 12761065 TI - 3D-Jury: a simple approach to improve protein structure predictions. AB - MOTIVATION: Consensus structure prediction methods (meta-predictors) have higher accuracy than individual structure prediction algorithms (their components). The goal for the development of the 3D-Jury system is to create a simple but powerful procedure for generating meta-predictions using variable sets of models obtained from diverse sources. The resulting protocol should help to improve the quality of structural annotations of novel proteins. RESULTS: The 3D-Jury system generates meta-predictions from sets of models created using variable methods. It is not necessary to know prior characteristics of the methods. The system is able to utilize immediately new components (additional prediction providers). The accuracy of the system is comparable with other well-tuned prediction servers. The algorithm resembles methods of selecting models generated using ab initio folding simulations. It is simple and offers a portable solution to improve the accuracy of other protein structure prediction protocols. AVAILABILITY: The 3D Jury system is available via the Structure Prediction Meta Server (http://BioInfo.PL/Meta/) to the academic community. SUPPLEMENTARY INFORMATION: 3D-Jury is coupled to the continuous online server evaluation program, LiveBench (http://BioInfo.PL/LiveBench/) PMID- 12761066 TI - Observing and interpreting correlations in metabolomic networks. AB - MOTIVATION: Metabolite profiling aims at an unbiased identification and quantification of all the metabolites present in a biological sample. Based on their pair-wise correlations, the data obtained from metabolomic experiments are organized into metabolic correlation networks and the key challenge is to deduce unknown pathways based on the observed correlations. However, the data generated is fundamentally different from traditional biological measurements and thus the analysis is often restricted to rather pragmatic approaches, such as data mining tools, to discriminate between different metabolic phenotypes. METHODS AND RESULTS: We investigate to what extent the data generated networks reflect the structure of the underlying biochemical pathways. The purpose of this work is 2 fold: Based on the theory of stochastic systems, we first introduce a framework which shows that the emergent correlations can be interpreted as a 'fingerprint' of the underlying biophysical system. This result leads to a systematic relationship between observed correlation networks and the underlying biochemical pathways. In a second step, we investigate to what extent our result is applicable to the problem of reverse engineering, i.e. to recover the underlying enzymatic reaction network from data. The implications of our findings for other bioinformatics approaches are discussed. PMID- 12761067 TI - Hierarchical analysis of dependency in metabolic networks. AB - MOTIVATION: Elucidation of metabolic networks for an increasing number of organisms reveals that even small networks can contain thousands of reactions and chemical species. The intimate connectivity between components complicates their decomposition into biologically meaningful sub-networks. Moreover, traditional higher-order representations of metabolic networks as metabolic pathways, suffers from the lack of rigorous definition, yielding pathways of disparate content and size. RESULTS: We introduce a hierarchical representation that emphasizes the gross organization of metabolic networks in largely independent pathways and sub systems at several levels of independence. The approach highlights the coupling of different pathways and the shared compounds responsible for those couplings. By assessing our results on Escherichia coli (E.coli metabolic reactions, Genetic Circuits Research Group, University of California, San Diego, http://gcrg.ucsd.edu/organisms/ecoli.html, 'model v 1.01. reactions') against accepted biochemical annotations, we provide the first systematic synopsis of an organism's metabolism. Comparison with operons of E.coli shows that low-level clusters are reflected in genome organization and gene regulation. AVAILABILITY: Source code, data sets and supplementary information are available at http://www.mas.ecp.fr/labo/equipe/gagneur/hierarchy/hierarchy.html PMID- 12761068 TI - Zerg: a very fast BLAST parser library. AB - SUMMARY: Zerg is a library of sub-routines that parses the output from all NCBI BLAST programs (Blastn, Blastp, Blastx, Tblastn and Tblastx) and returns the attributes of a BLAST report to the user. It is optimized for speed, being especially useful for large-scale genomic analysis. Benchmark tests show that Zerg is over two orders of magnitude faster than some widely used BLAST parsers. AVAILABILITY: http://bioinfo.iq.usp.br/zerg PMID- 12761069 TI - Visual representation of database search results: the RHIMS Plot. AB - SUMMARY: An algorithm and software are described that provide a fast method to produce a novel, function-oriented visualization of the results of a sequence database search. Text mining of sequence annotations allows position specific plots of potential functional similarity to be compared in a simple compact representation. AVAILABILITY: The application can be accessed via a web server at http://www.compbio.dundee.ac.uk. The RHIMS software may be obtained by request to the authors. PMID- 12761070 TI - ACGT-a comparative genomics tool. AB - SUMMARY: ACGT (a comparative genomics tool) is a genomic DNA sequence comparison viewer and analyzer. It can read a pair of DNA sequences in GenBank, Embl or Fasta formats, with or without a comparison file, and provide users with many options to view and analyze the similarities between the input sequences. It is written in Java and can be run on Unix, Linux and Windows platforms. AVAILABILITY: The ACGT program is freely available with documentation and examples at website: http://db.systemsbiology.net/projects/local/mhc/acgt/ PMID- 12761071 TI - Ligand-Info, searching for similar small compounds using index profiles. AB - MOTIVATION: The Ligand-Info system is based on the assumption that small molecules with similar structure have similar functional (binding) properties. The developed system enables a fast and sensitive index based search for similar compounds in large databases. Index profiles, constructed by averaging indexes of related molecules are used to increase the specificity of the search. The utilization of index profiles helps to focus on frequent, common features of a family of compounds. RESULTS: A Java-based tool for clustering and scanning of small molecules has been created. The tool can interactively cluster sets of molecules and create index profiles on the user side and automatically download similar molecules from a databases of 250 000 compounds. The results of the application of index profiles demonstrate that the profile based search strategy can increase the quality of the selection process. AVAILABILITY: The system is available at http://Ligand.Info. The application requires the Java Runtime Environment 1.4, which can be automatically installed during the first use on desktop systems, which support it. A standalone version of the program is available from the authors upon request. PMID- 12761072 TI - CAP: conformation angles package-displaying the conformation angles of side chains in proteins. AB - SUMMARY: A graphics package has been developed to display all side chain conformation angles of the user selected residue in a given protein structure. The proposed package is incorporated with all the protein structures (solved using X-ray diffraction and NMR spectroscopy) available in the Protein Data Bank. The package displays the multiple conformations adopted by a single amino acid residue whose structure is solved and refined at very high resolution. In addition, it shows the percentage distribution of the side chain conformation angles in different rotameric states. AVAILABILITY: http://144.16.71.146/cap/ PMID- 12761073 TI - Bacteriophage and the evolution of epidemic cholera. PMID- 12761074 TI - Extracellular enzymes with immunomodulating activities: variations on a theme in Streptococcus pyogenes. PMID- 12761075 TI - Examination of diverse toxin-coregulated pilus-positive Vibrio cholerae strains fails to demonstrate evidence for Vibrio pathogenicity island phage. AB - The major virulence factors of toxigenic Vibrio cholerae are cholera toxin, which is encoded by a lysogenic filamentous bacteriophage (CTXPhi), and toxin coregulated pilus (TCP), an essential colonization factor that is also the receptor for CTXPhi. The genes involved in the biosynthesis of TCP reside in a pathogenicity island, which has been reported to correspond to the genome of another filamentous phage (designated VPIPhi) and to encode functions necessary for the production of infectious VPIPhi particles. We examined 46 V. cholerae strains having diverse origins and carrying different genetic variants of the TCP island for the production of the VPIPhi and CTXPhi in different culture conditions, including induction of prophages with mitomycin C and UV irradiation. Although 9 of 10 V. cholerae O139 strains and 12 of 15 toxigenic El Tor strains tested produced extracellular CTXPhi, none of the 46 TCP-positive strains produced detectable VPIPhi in repeated assays, which detected as few as 10 particles of a control CTX phage per ml. These results contradict the previous report regarding VPIPhi-mediated horizontal transfer of the TCP genes and suggest that the TCP island is unable to support the production of phage particles. Further studies are necessary to understand the mechanism of horizontal transfer of the TCP island. PMID- 12761076 TI - Mammalian transforming growth factor beta1 activated after ingestion by Anopheles stephensi modulates mosquito immunity. AB - During the process of bloodfeeding by Anopheles stephensi, mammalian latent transforming growth factor beta1 (TGF-beta1) is ingested and activated rapidly in the mosquito midgut. Activation may involve heme and nitric oxide (NO), agents released in the midgut during blood digestion and catalysis of L-arginine oxidation by A. stephensi NO synthase (AsNOS). Active TGF-beta1 persists in the mosquito midgut to extended times postingestion and is recognized by mosquito cells as a cytokine. In a manner analogous to the regulation of vertebrate inducible NO synthase and malaria parasite (Plasmodium) infection in mammals by TGF-beta1, TGF-beta1 regulates AsNOS expression and Plasmodium development in A. stephensi. Together, these observations indicate that, through conserved immunological cross talk, mammalian and mosquito immune systems interface with each other to influence the cycle of Plasmodium development. PMID- 12761077 TI - A recombinant beta-1,3-glucanosyltransferase homolog of Coccidioides posadasii protects mice against coccidioidomycosis. AB - Coccidioides posadasii is a fungal respiratory pathogen which is responsible for recurrent epidemics of San Joaquin Valley fever (coccidioidomycosis) in desert regions of the southwestern United States. Numerous studies have revealed that the cell wall of the parasitic phase of the fungus is a reservoir of immunoreactive macromolecules and a potential source of a vaccine against this mycosis. A 495-bp fragment of a C. posadasii gene which encodes a putative wall associated, glycosylphosphatidylinositol (GPI)-anchored beta-1,3 glucanosyltransferase was identified by computational analysis of the partially sequenced genome of this pathogen. The translated, full-length gene (GEL1) showed high sequence homology to a reported beta-1,3-glucanosyltransferase of Aspergillus fumigatus (70% identity, 90% similarity) and was selected for further study. The GEL1 mRNA of C. posadasii was detected at the highest level during the endosporulation stage of the parasitic cycle, and the mature protein was immunolocalized to the surface of endospores. BALB/c or C57BL/6 mice were immunized subcutaneously with the bacterium-expressed recombinant protein (rGel1p) to evaluate its protective efficacy against a lethal challenge of C. posadasii by either the intraperitoneal or intranasal route. In both cases, rGel1p-immune mice infected with the pathogen showed a significant reduction in fungal burden and increased survival compared to nonimmune mice. The recombinant beta-1,3-glucanosyltransferase is a valuable addition to an arsenal of immunoreactive proteins which could be incorporated into a human vaccine against coccidioidomycosis. PMID- 12761078 TI - Isolation and characterization of mini-Tn5Km2 insertion mutants of Brucella abortus deficient in internalization and intracellular growth in HeLa cells. AB - Brucella spp. are facultative intracellular pathogens that have the ability to survive and multiply in professional and nonprofessional phagocytes and cause abortion in domestic animals and undulant fever in humans. The mechanism and factors of virulence are not fully understood. To identify genes related to internalization and multiplication in host cells, Brucella abortus was mutagenized by mini-Tn5Km2 transposon that carryied the kanamycin resistance gene, 4,400 mutants were screened, and HeLa cells were infected with each mutant. Twenty-three intracellular-growth-defective mutants were screened and were characterized for internalization and intracellular growth. From these results, we divided the mutants into the following three groups: class I, no internalization and intracellular growth within HeLa cells; class II, an internalization similar to that of the wild type but with no intracellular growth; and class III, internalization twice as high as the wild type but with no intracellular growth. Sequence analysis of DNA flanking the site of transposon showed various insertion sites of bacterial genes that are virulence-associated genes, including virB genes, an ion transporter system, and biosynthesis- and metabolism-associated genes. These internalization and intracellular-growth defective mutants in HeLa cells also showed defective intracellular growth in macrophages. These results suggest that the virulence-associated genes isolated here contributed to the intracellular growth of both nonprofessional and professional phagocytes. PMID- 12761079 TI - A fluoroquinolone induces a novel mitogen-encoding bacteriophage in Streptococcus canis. AB - This study investigated whether the recently recognized emergence of canine streptococcal toxic shock syndrome (STSS) and necrotizing fasciitis (NF) might be partly attributed to the use of fluoroquinolones to treat Streptococcus canis infections in dogs. Both mitomycin and the fluoroquinolone enrofloxacin caused bacteriophage-induced lysis of S. canis strain 34, an isolate from a case of canine STSS and NF. Fluoroquinolone-evoked, bacteriophage-induced lysis occurred over a range of concentrations similar to those that would occur after treatment of dogs with these agents. To search for a possible bacteriophage-encoded streptococcal superantigen gene(s), a library of the 36.5 (+/-1.1)-kb bacteriophage, designated phisc1, was made by ligating 3- to 7-kb Tsp5091 digested phisc1 fragments into an EcoRI-digested lambdaZapII vector. Recombinants were screened for mitogenic activity by using canine peripheral blood lymphocytes. Of 800 recombinants screened, 11 recombinants with mitogenic effects were identified, and their inserts were sequenced. The highest homology of 11.6 kb of sequenced phisc1 DNA was to the completely sequenced Streptococcus pneumoniae bacteriophage MM1. Seven of the 11 phisc1 sequenced inserts contained a 552-bp open reading frame, scm, with 27% amino acid similarity to pokeweed (Phytolacca americana) mitogen. PCR showed this gene to be present in 22 of 23 S. canis isolates tested. Quantitative reverse transcription-PCR showed that bacteriophage induction was associated with a 58-fold enhancement of expression of this gene relative to that in a noninduced culture of a similar age. The presence of this gene on a fluoroquinolone-induced bacteriophage may explain the association observed between fluoroquinolone use in dogs and the development of canine STTS and NF. PMID- 12761080 TI - Killing of Aspergillus fumigatus by alveolar macrophages is mediated by reactive oxidant intermediates. AB - Phagocytosis and mechanisms of killing of Aspergillus fumigatus conidia by murine alveolar macrophages (AM), which are the main phagocytic cells of the innate immunity of the lung, were investigated. Engulfment of conidia by murine AM lasts 2 h. Killing of A. fumigatus conidia by AM begins after 6 h of phagocytosis. Swelling of the conidia inside the AM is a prerequisite for killing of conidia. The contributions of NADPH oxidase and inducible nitric oxide synthase to the conidicidal activity of AM were studied using AM from OF1, wild-type and congenic p47phox(-/-) 129Sv, and wild-type and congenic iNOS(-/-) C57BL/6 mice. AM from p47phox(-/-) mice were unable to kill A. fumigatus conidia. Inhibitors of NADPH oxidase that decreased the production of reactive oxidant intermediates inhibited the killing of A. fumigatus without altering the phagocytosis rate. In contrast to NADPH oxidase, nitric oxide synthase does not play a role in killing of conidia. Corticosteroids did not alter the internalization of conidia by AM but did inhibit the production of reactive oxidant intermediates and the killing of A. fumigatus conidia by AM. Impairment of production of reactive oxidant intermediates by corticosteroids is responsible for the development of invasive aspergillosis in immunosuppressed mice. PMID- 12761081 TI - Comparison of bipA alleles within and across Bordetella species. AB - The Bordetella BvgAS signal transduction system controls the expression of at least three phenotypic phases, the Bvg(+) or virulent phase, the Bvg(-) or avirulent phase, and the Bvg(i) or Bvg intermediate phase, which has been hypothesized to be important for transmission. bipA, the first identified Bvg(i) phase gene, encodes a protein with similarity to the well-characterized bacterial adhesins intimin and invasin. Proteins encoded by the bipA genes present in Bordetella pertussis Tohama I and Bordetella bronchiseptica RB50 differ in the number of 90-amino-acid repeats which they possess and in the sequence of the C terminal domain. To investigate the possibility that bipA alleles segregate according to host specificity and to gain insight into the role of BipA and the Bvg(i) phase in the Bordetella infectious cycle, we compared bipA alleles across members of the B. bronchiseptica cluster, which includes both human-infective (B. pertussis and B. parapertussis(hu)) and non-human-infective (B. bronchiseptica and B. parapertussis(ov)) strains. bipA genes were present in most, but not all, strains. All bipA genes present in B. bronchiseptica strains were identical to bipA of RB50 (at least with regard to the DNA sequence of the 3' C-terminal domain-encoding region, the number of 90-amino-acid repeats encoded, and expression patterns). Although all bipA genes present in the other Bordetella strains were identical in the 3' C-terminal-domain-encoding region to bipA of B. pertussis Tohama I, they varied in the number of 90-amino-acid repeats that they encoded and in expression level. Notably, the genes present in B. parapertussis(hu) strains were pseudogenes, and the genes present in B. parapertussis(ov) strains were expressed at significantly reduced levels compared with the levels in B. pertussis and B. bronchiseptica strains. Our results indicate that there is a correlation between specific bipA alleles and specific hosts. They also support the hypothesis that both horizontal gene transfer and fine-tuning of gene expression patterns contribute to the evolution of host adaptation in lineages of the B. bronchiseptica cluster. PMID- 12761082 TI - Involvement of beta interferon in enhancing inducible nitric oxide synthase production and antimicrobial activity of Burkholderia pseudomallei-infected macrophages. AB - Burkholderia pseudomallei is the causative agent of melioidosis, a life threatening disease that affects both humans and animals. This bacterium is able to survive and multiply inside both phagocytic and nonphagocytic cells. We recently reported that mouse macrophages infected with B. pseudomallei fail to produce a significant level of inducible nitric oxide synthase (iNOS), a crucial enzyme needed for the cells to control the intracellular growth of this bacterium. In the present study, we extended our investigation to demonstrate that, unlike other gram-negative bacteria that have been investigated, B. pseudomallei only minimally activates beta interferon (IFN-beta) production; this minimal activation leads to a low level of interferon regulating factor 1 (IRF-1) in the macrophages, in parallel with poor iNOS expression. Adding exogenous IFN beta to the system could upregulate IRF-1 production, which in turn could enhance iNOS expression in the B. pseudomallei-infected macrophages and lead to suppression of the intracellular growth of this bacterium. Taken together, these results imply that the failure of macrophages to successfully control the growth and survival of intracellular B. pseudomallei is related, at least in part, to the defective production of IFN-beta, which modulates the ability of macrophages to synthesize iNOS. PMID- 12761083 TI - Stimulation of hematopoiesis by the Fms-like tyrosine kinase 3 ligand restores bacterial induction of Th1 cytokines in thermally injured mice. AB - Patients with large burn injuries are susceptible to opportunistic infections due to impaired functions of multiple effector cells of innate immunity and acquired immunity, including macrophages, dendritic cells (DC), natural killer (NK) cells, and T cells. The ability of a host to produce Th1 cytokines, such as gamma interferon (IFN-gamma) and interleukin-12 (IL-12), upon infectious challenge is also impaired after burn injury. Stimulation of hematopoiesis, to regenerate new immune cells, may be an effective strategy for improving resistance to infections after severe burn trauma. Fms-like tyrosine kinase 3 ligand (Flt3L) is a hematopoietic cytokine that stimulates the expansion and differentiation of NK cells and DC. Using a mouse model, we tested the hypothesis that Flt3L treatments after burn injury stimulate the production of functional effector cells of innate immunity and restore appropriate Th1 cytokine responses to Pseudomonas aeruginosa, a common source of pneumonia and wound infections in burn victims. Flt3L increased splenic cellularity in sham (uninjured) and burned mice and increased the numbers of NK cells (DX5(+)) and DC (CD11c(+)). In response to P. aeruginosa, significant increases in the serum IFN-gamma levels and the numbers of splenic IFN-gamma-producing DC, NK cells, and T cells were observed in Flt3L treated burned mice compared to the values obtained for untreated burned mice. The splenic levels of IL-12 and IL-15 mRNAs and the IL-12 and IL-15 receptors were also increased. In addition, Flt3L treatment restored the ability of splenic cultures prepared from burned mice to produce IFN-gamma and IL-12 after in vitro challenge with P. aeruginosa. Flt3L may have potential for restoring NK cell and DC functions and improving immunity after burn injury. PMID- 12761084 TI - Role of phosphoglucomutase of Stenotrophomonas maltophilia in lipopolysaccharide biosynthesis, virulence, and antibiotic resistance. AB - A homologue of the algC gene, responsible for the production of a phosphoglucomutase (PGM) associated with LPS and alginate biosynthesis in Pseudomonas aeruginosa, spgM, was cloned from Stenotrophomonas maltophilia. The spgM gene was shown to encode a bifunctional enzyme with both PGM and phosphomannomutase activities. Mutants lacking spgM produced less LPS than the SpgM(+) parent strain and had a tendency for shorter O polysaccharide chains. No changes in LPS chemistry were obvious as a result of the loss of spgM. Significantly, however, spgM mutants displayed a modest increase in susceptibility to several antimicrobial agents and were completely avirulent in an animal model of infection. The latter finding may relate to the resultant serum sensitivity of spgM mutants which, unlike the wild-type parent strain, were rapidly killed by human serum. These data highlight the contribution made by LPS to the antimicrobial resistance and virulence of S. maltophilia. PMID- 12761085 TI - T-cell responses to CD1-presented lipid antigens in humans with Mycobacterium tuberculosis infection. AB - CD1-restricted presentation of lipid or glycolipid antigens derived from Mycobacterium tuberculosis has been demonstrated by in vitro experiments using cultured T-cell lines. In the present work, the frequency of T-cell responses to natural mycobacterial lipids was analyzed in ex vivo studies of peripheral blood lymphocytes from human patients with pulmonary tuberculosis, from asymptomatic individuals with known contact with M. tuberculosis documented by conversion of their tuberculin skin tests, and from healthy tuberculin skin test-negative individuals or individuals vaccinated with Mycobacterium bovis BCG. Proliferation and gamma interferon enzyme-linked immunospot assays using peripheral blood lymphocytes and autologous CD1(+) immature dendritic cells revealed that T cells from asymptomatic M. tuberculosis-infected donors responded with significantly greater magnitude and frequency to mycobacterial lipid antigen preparations than lymphocytes from uninfected healthy donors. By use of these methods, lipid antigen-specific proliferative responses were minimally detectable or absent in blood samples from patients with active tuberculosis prior to chemotherapy but became detectable in blood samples drawn 2 weeks after the start of treatment. Lipid antigen-reactive T cells were detected predominantly in the CD4-enriched T cell fractions of circulating lymphocytes, and anti-CD1 antibody blocking experiments confirmed the CD1 restriction of these T-cell responses. Our results provide further support for the hypothesis that lipid antigens serve as targets of the recall response to M. tuberculosis, and they indicate that CD1-restricted T cells responding to these antigens comprise a significant portion of the circulating pool of M. tuberculosis-reactive T cells in healthy individuals with previous exposure to M. tuberculosis. PMID- 12761086 TI - DegS is necessary for virulence and is among extraintestinal Escherichia coli genes induced in murine peritonitis. AB - Extraintestinal Escherichia coli strains cause meningitis, sepsis, urinary tract infection, and other infections outside the bowel. We examined here extraintestinal E. coli strain CFT073 by differential fluorescence induction. Pools of CFT073 clones carrying a CFT073 genomic fragment library in a promoterless gfp vector were inoculated intraperitoneally into mice; bacteria were recovered by lavage 6 h later and then subjected to fluorescence-activated cell sorting. Eleven promoters were found to be active in the mouse but not in Luria-Bertani (LB) broth culture. Three are linked to genes for enterobactin, aerobactin, and yersiniabactin. Three others are linked to the metabolic genes metA, gltB, and sucA, and another was linked to iha, a possible adhesin. Three lie before open reading frames of unknown function. One promoter is associated with degS, an inner membrane protease. Mutants of the in vivo-induced loci were tested in competition with the wild type in mouse peritonitis. Of the mutants tested, only CFT073 degS was found to be attenuated in peritoneal and in urinary tract infection, with virulence restored by complementation. CFT073 degS shows growth similar to that of the wild type at 37 degrees C but is impaired at 43 degrees C or in 3% ethanol LB broth at 37 degrees C. Compared to the wild type, the mutant shows similar serum survival, motility, hemolysis, erythrocyte agglutination, and tolerance to oxidative stress. It also has the same lipopolysaccharide appearance on a silver-stained gel. The basis for the virulence attenuation is unclear, but because DegS is needed for sigma(E) activity, our findings implicate sigma(E) and its regulon in E. coli extraintestinal pathogenesis. PMID- 12761087 TI - T cells are required for host protection against Brugia malayi but need not produce or respond to interleukin-4. AB - T cells are known to be required for host protection in mouse models of Brugia malayi infection. Several independent studies in murine models have also highlighted the rapid induction of Th2-like responses after infection with B. malayi or B. pahangi. Previous data from our laboratory have described a significant increase in permissiveness in the absence of interleukin-4 (IL-4), the "prototypical" Th2 cytokine, involved in both the induction and maintenance of Th2 responses. These observations led to our hypothesis that T cells involved in murine host protection would respond to IL-4 signaling and differentiate into cells of the "type 2" phenotype. As such, these cells would presumably also act as major sources of IL-4. To investigate these hypotheses, we performed several adoptive transfers in which we controlled the cell population(s) able to produce or respond to IL-4. We show here that, in contrast to our original hypotheses, IL 4 production and IL-4 receptor expression by T cells are both dispensable for T cell-mediated host protection. Instead, our data imply that T cells may be required for eosinophil accumulation at the site of infection. PMID- 12761088 TI - Nonpathogenic Escherichia coli can contribute to the production of Shiga toxin. AB - The food-borne pathogen, Escherichia coli O157:H7, has been associated with gastrointestinal disease and the life-threatening sequela hemolytic uremic syndrome. The genes for the virulence factor, Shiga toxin 2 (Stx2), in E. coli O157:H7 are encoded on a temperate bacteriophage under the regulation of the late gene promoter. Induction of the phage lytic cycle is required for toxin synthesis and release. We investigated the hypothesis that nonpathogenic E. coli could amplify Stx2 production if infected with the toxin-encoding phage. Toxin-encoding phage were incubated with E. coli that were either susceptible or resistant to the phage. The addition of phage to phage-susceptible bacteria resulted in up to 40-fold more toxin than a pure culture of lysogens, whereas the addition of phage to phage-resistant bacteria resulted in significantly reduced levels of toxin. Intestinal E. coli isolates incubated with Shiga toxin-encoding phage produced variable amounts of toxin. Of 37 isolates, 3 produced significantly more toxin than was present in the inoculum, and 1 fecal isolate appeared to inactivate the toxin. Toxin production in the intestine was assessed in a murine model. Fecal toxin recovery was significantly reduced when phage-resistant E. coli was present. These results suggest that the susceptibility of the intestinal flora to the Shiga toxin phage could exert either a protective or an antagonistic influence on the severity of disease by pathogens with phage-encoded Shiga toxin. Toxin production by intestinal flora may represent a novel strategy of pathogenesis. PMID- 12761089 TI - Relationship of plcR-regulated factors to Bacillus endophthalmitis virulence. AB - The explosive, destructive course of Bacillus endophthalmitis has been attributed to the production of toxins during infection. In this study we analyzed the contribution of toxins controlled by the global regulator plcR to the pathogenesis of experimental Bacillus endophthalmitis. Isogenic plcR-deficient mutants of Bacillus cereus and Bacillus thuringiensis were constructed by insertional inactivation of plcR by the kanamycin resistance cassette, aphA3. Rabbit eyes were injected intravitreally with approximately 100 CFU of wild-type B. cereus or B. thuringiensis or a plcR-deficient mutant. The evolution of endophthalmitis resulting from each plcR-deficient mutant was considerably slower than that caused by each wild-type strain. Retinal function was not eliminated until 42 h postinfection in rabbits with endophthalmitis caused by the plcR deficient mutants, whereas wild-type infections resulted in a complete loss of retinal function within 18 h. The intraocular inflammatory cell influx and retinal destruction in plcR-deficient endophthalmitis approached the severity observed in wild-ype infections, but not until 36 h postinfection. Gross and histological examinations of eyes infected with plcR mutants demonstrated that the anterior and posterior segment changes were muted compared to the changes observed in eyes infected with the wild types. The loss of plcR-regulated factors significantly attenuated the severity of Bacillus endophthalmitis. The results therefore suggest that plcR may represent a target for which adjunct therapies could be designed for the prevention of blindness during Bacillus endophthalmitis. PMID- 12761090 TI - Stx2-specific human monoclonal antibodies protect mice against lethal infection with Escherichia coli expressing Stx2 variants. AB - Shiga toxin-producing Escherichia coli (STEC) strains are responsible for causing hemolytic-uremic syndrome (HUS), and systemic administration of Shiga toxin (Stx) specific human monoclonal antibodies (HuMAbs) is considered a promising approach for prevention or treatment of the disease in children. The goal of the present study was to investigate the ability of Stx2-specific HuMAbs to protect against infections with STEC strains that produce Stx2 variants. Dose-response studies on five HuMAbs, using the mouse toxicity model, revealed that only the three directed against the A subunit were protective against Stx2 variants, and 5C12 was the most effective among the three tested. Two HuMAbs directed against the B subunit, while highly effective against Stx2, were ineffective against Stx2 variants. In a streptomycin-treated mouse model, parenteral administration of 5C12 significantly protected mice up to 48 h after oral bacterial challenge. We conclude that 5C12, reactive against the Stx2 A subunit, is an excellent candidate for immunotherapy against HUS and that antibodies directed against the A subunit of Stx2 have broad-spectrum activity that includes Stx2 variants, compared with those directed against the B subunit. PMID- 12761092 TI - Plasmid stability during in vitro propagation of Borrelia burgdorferi assessed at a clonal level. AB - Borrelia burgdorferi causes Lyme disease in humans. The genome of the sequenced type strain B31 MI consists of a linear chromosome, 12 linear plasmids, and 9 circular plasmids. Previous studies by other investigators indicated that some of these plasmids are essential for the survival of the spirochetes in vivo but not in vitro. We have studied plasmid stability during in vitro growth at 23 and 35 degrees C, conditions that approximate the temperatures of the tick vector and the mammalian host, respectively. Starting with two clones that have all 21 plasmids, we investigated plasmid maintenance within the population and on a clonal level. After three passages (27 generations), the cultures were no longer homogeneous and some derivative clones had already lost multiple plasmids. Despite this, one of six clones analyzed after 25 passages (225 generations) retained all but one plasmid (cp9) and was able to complete the mouse-tick-mouse infectious cycle. We analyzed protein composition and regulation of gene expression of clones differing in plasmid content after serial passages. All clones tested exhibited temperature-regulated expression of several proteins, including OspC. In addition, analysis of cultures inoculated from frozen stocks suggests that freezing and/or thawing contributes to heterogeneity in the outgrowth population with respect to plasmid content. Our investigations show that in vitro propagation of a clone leads to a heterogeneous population but that virulent clones can persist through extended passage. We therefore conclude that isogenicity of clones must be confirmed irrespective of their in vitro passage history. PMID- 12761091 TI - Critical contribution of CD28-CD80/CD86 costimulatory pathway to protection from Trypanosoma cruzi infection. AB - The CD28-CD80/CD86-mediated T-cell costimulatory pathway has been variably implicated in infectious immunity. In this study, we investigated the role of this costimulatory pathway in resistance to Trypanosoma cruzi infection by using CD28-deficient mice and blocking antibodies against CD80 and CD86. CD28-deficient mice exhibited markedly exacerbated T. cruzi infection, as evidenced by unrelenting parasitemia and 100% mortality after infection with doses that are nonlethal in wild-type mice. The blockade of both CD80 and CD86 by administering specific monoclonal antibodies also exacerbated T. cruzi infection in wild-type mice. Splenocytes from T. cruzi-infected, CD28-deficient mice exhibited greatly impaired gamma interferon production in response to T. cruzi antigen stimulation in vitro compared to those from infected wild-type mice. The induction of T. cruzi antigen-specific CD8(+) T cells was also impaired in T. cruzi-infected, CD28-deficient mice. In addition to these defects in natural protection against T. cruzi infection, CD28-deficient mice were also defective in the induction of CD8(+)-T-cell-mediated protective immunity against T. cruzi infection by DNA vaccination. These results demonstrate, for the first time, a critical contribution of the CD28-CD80/CD86 costimulatory pathway not only to natural protection against primary T. cruzi infection but also to DNA vaccine-induced protective immunity to Chagas' disease. PMID- 12761093 TI - The Mycobacterium tuberculosis recombinant 27-kilodalton lipoprotein induces a strong Th1-type immune response deleterious to protection. AB - Th1 immune response is essential in the protection against mycobacterial intracellular pathogens. Lipoproteins trigger both humoral and cellular immune responses and may be candidate protective antigens. We studied in BALB/c mice the immunogenicity and the protection offered by the recombinant 27-kDa Mycobacterium tuberculosis lipoprotein and the corresponding DNA vaccine. Immunization with the 27-kDa antigen resulted in high titers of immunoglobulin G1 (IgG1) and IgG2a with a typical Th1 profile and a strong delayed hypersensitivity response. A strong proliferation response was observed in splenocytes, and significant nitric oxide production and gamma interferon secretion but not interleukin 10 secretion were measured. Based on these criteria, the 27-kDa antigen induced a typical Th1-type immune response thought to be necessary for protection. Surprisingly, in 27-kDa vaccinated mice (protein or DNA vaccines) challenged by M. tuberculosis H37Rv or BCG strains, there was a significant increase in the numbers of CFU in the spleen compared to that for control groups. Furthermore, the protection provided by BCG or other mycobacterial antigens was completely abolished once the 27-kDa antigen was added to the vaccine preparations. This study indicates that the 27-kDa antigen has an adverse effect on the protection afforded by recognized vaccines. We are currently studying how the 27-kDa antigen modulates the mouse immune response. PMID- 12761094 TI - Optimal tumor necrosis factor induction by Plasmodium falciparum requires the highly localized release of parasite products. AB - Overproduction of tumor necrosis factor (TNF) has been linked with the pathogenesis of Plasmodium falciparum malaria. Here, we examined why the high levels of TNF-inducing activity associated with P. falciparum-parasitized erythrocytes (PE) appear to be lost after cell lysis. Static coculture of PE and peripheral blood mononuclear cells (PBMC), with or without separation by porous membranes, demonstrated that rupture of live PE in the presence of responder cells was required for optimal TNF induction. Although the insoluble fraction of lysed PE was found to partially inhibit TNF responses, supernatants prepared from large numbers of lysed PE still contained only low levels of TNF-inducing activity, which showed no evidence of instability. A dramatic reduction in TNF levels resulted when noncytoadherent PE lines were maintained under low-cell proximity conditions by suspension coculture. This reduction was much less marked with PE capable of adhering to PBMC, despite the fact that cytoadherent and noncytoadherent parasite lines induced comparable levels of TNF in high-cell proximity, static coculture. These results suggest that rupture of PE in a highly localized setting, facilitated by either static coculture or the more biologically relevant phenomenon of cytoadherence to PBMC, can result in considerable enhancement of the P. falciparum-induced TNF response. PMID- 12761095 TI - Humoral and cellular immune responses to Trypanosoma cruzi-derived paraflagellar rod proteins in patients with Chagas' disease. AB - Sera and peripheral blood mononuclear cells (PBMC) from patients displaying different clinical symptoms as well as from normal uninfected individuals (NI) were used to evaluate the humoral and cellular responses of Chagas' disease patients to Trypanosoma cruzi-derived paraflagellar rod proteins (PFR). Our results show that sera from both asymptomatic Chagas' disease patients (ACP) and cardiac Chagas' disease patients (CCP) have higher levels of antibodies to PFR than sera from NI. Immunoglobulin G1 (IgG1) and IgG3 were the main Ig isotypes that recognized PFR. We also tested three recombinant forms of PFR, named rPAR-1, rPAR-2, and rPAR-3, by Western blot analysis. Sera from seven out of eight patients with Chagas' disease recognized one of the three rPAR forms. Sera from 75, 50, and 37.5% of Chagas' disease patients tested recognized rPAR-3, rPAR-2, and rPAR-1, respectively. PFR induced proliferation of 100 and 70% of PBMC from ACP and CCP, respectively. Further, stimulation of cells from Chagas' disease patients with PFR enhanced the frequencies of both small and large CD4(+) CD25(+) and CD4(+) CD69(+) lymphocytes, as well as that of small CD8(+) CD25(+) lymphocytes. Finally, we evaluated the ability of PFR to elicit the production of gamma interferon (IFN-gamma) by PBMC from patients with Chagas' disease. Fifty percent of the PBMC from ACP as well as CCP produced IFN-gamma upon stimulation with PFR. PFR enhanced the percentages of IFN-gamma-producing cells in both CD3(+) and CD3(-) populations. Within the T-cell population, large CD4(+) T lymphocytes were the main source of IFN-gamma. PMID- 12761096 TI - Perforin and gamma interferon are critical CD8+ T-cell-mediated responses in vaccine-induced immunity against Leishmania amazonensis infection. AB - Previous studies have demonstrated that protection against New World leishmaniasis caused by Leishmania amazonensis can be elicited by immunization with the developmentally regulated Leishmania amastigote antigen, P-8. In this study, several independent experimental approaches were employed to investigate the protective immunological mechanisms involved. T-cell subset depletion experiments clearly indicate that elicitation of CD8(+) (as well as CD4(+)) effector responses is required for protection. Further, mice lacking beta(2) microglobulin (and hence deficient in major histocompatibility complex class I antigen presentation) were not able to control a challenge infection after vaccination, indicating an essential protective role for CD8(+) T effector responses. Analysis of the events ongoing at the cutaneous site of infection indicated a changing cellular dynamic involved in protection. Early postinfection in protectively vaccinated mice, a predominance of CD8(+) T cells, secreting gamma interferon (IFN-gamma) and expressing perforin, was observed at the site of infection; subsequently, activated CD4(+) T cells producing IFN-gamma were primarily found. As protection correlated with the ratio of total IFN-gamma producing cells (CD4(+) and CD8(+) T cells) to macrophages found at the site of infection, a role for IFN-gamma was evident; in addition, vaccination of IFN gamma-deficient mice failed to provide protection. To further assess the effector mechanisms that mediate protection, mice deficient in perforin synthesis were examined. Perforin-deficient mice vaccinated with the P-8 antigen were unable to control infection. Thus, the elicitation of CD8(+) T cell effector mechanisms (perforin, IFN-gamma) are clearly required in the protective immune response against L. amazonensis infection in vaccinated mice. PMID- 12761098 TI - Expression of multiple CPB genes encoding cysteine proteases is required for Leishmania mexicana virulence in vivo. AB - Leishmania mexicana mutants deficient in the multicopy CPB gene array have reduced virulence, demonstrated by poor lesion growth in BALB/c mice and induction of a protective Th1 response. Reinsertion of the amastigote-specific CPB2.8 or metacyclic stage-specific CPB2 gene into a CPB-deficient mutant L. mexicana failed to restore either a Th2 response or sustained virulence. However, reexpression of multiple CPB genes from a cosmid significantly restored virulence. This was characterized by increased lesion and parasite growth and the acquisition of a Th2 response, as determined by measuring interleukin-4 production and immunoglobulin G1 (IgG1) and IgE levels. These studies confirm that L. mexicana cysteine proteases are important virulence factors and provide an explanation for the presence in L. mexicana of a multicopy tandem array of CPB genes. PMID- 12761097 TI - Characterization of anthrolysin O, the Bacillus anthracis cholesterol-dependent cytolysin. AB - We characterized the expression of a putative toxin of Bacillus anthracis, a member of the cholesterol-dependent cytolysin (CDC) family, which includes listeriolysin O, perfringolysin O, and streptolysin O. We named this cytotoxin anthrolysin O (ALO). Although B. anthracis expresses minimal hemolytic activity in clinical settings, we show that Sterne strain 7702 expresses hemolytic activity when grown in brain heart infusion broth or in other rich bacteriologic media, but it secretes barely detectable amounts of hemolysin when grown in Luria Bertani (LB) broth. Glucose supplementation of LB broth increases the amount of secreted hemolytic activity. Expression of hemolytic activity is maximal during mid- to late-log phase and decreases in the stationary phase. These observations are supported, in part, by semiquantitative reverse transcriptase PCR of alo mRNA. Hemolytic activity in growth supernatants was increased in the presence of reducing agent and almost totally inhibited in a dose-dependent manner by cholesterol; both of these activities are characteristic of a CDC toxin. A mutant of Sterne strain 7702, strain UT231, in which the alo gene was deleted and replaced by a kanamycin cassette, secreted barely detectable hemolytic activity into the growth medium. When strain UT231 was complemented in trans with native alo on a low-copy-number plasmid [strain UT231(pUTE554)], it regained the ability to secrete hemolytic activity, indicating that ALO is the major hemolysin secreted by this strain of B. anthracis in rich media in vitro. To further support the alo gene product being a hemolysin, recombinant B. anthracis ALO (rALO) purified from Escherichia coli was extremely active against washed human erythrocytes, with complete hemolysis detected at approximately 30 molecules of rALO per erythrocyte. Considering the virulence roles of CDCs for other gram positive bacteria, we speculate that ALO may have a role in anthrax virulence. PMID- 12761099 TI - Modulation of virulence by two acidified nitrite-responsive loci of Salmonella enterica serovar Typhimurium. AB - Two acidified nitrite-inducible genes of Salmonella enterica serovar Typhimurium were identified with a green fluorescent protein-based promoter-trap screen. The nitrite-inducible promoters were located upstream of loci that we designated nipAB and nipC, which correspond to hcp-hcr (hybrid cluster protein) of Escherichia coli and norA of Alcaligenes eutrophus, respectively. Maximal induction of the promoters by nitrite was dependent on pH. The nipAB promoter was regulated by oxygen in an Fnr-dependent manner. The nipC promoter was also regulated by oxygen but in an Fnr-independent manner. The promoters were upregulated in activated RAW264.7 macrophage-like cells, which produce NO via the inducible nitric oxide synthase (iNOS), and the induction was inhibited by aminoguanidine, an inhibitor of iNOS. Although the nipAB and nipC mutants displayed no defects under a variety of in vitro conditions or in tissue culture infections, they exhibited lower oral 50% lethal doses (LD(50)s) than did the wild type in C57BL/6J mouse infections. The lower LD(50)s reflected an unexpected increased ability of small inoculating doses of the mutant bacteria to cause lethal infection 2 to 3 weeks after challenge, compared to a similar challenge dose of wild-type bacteria. We conclude that these genes are regulated by physiological nitrogen oxides and that the absence of these bacterial genes in some way diminishes the ability of mice to clear a low dose infection. PMID- 12761100 TI - Mutation of luxS of Streptococcus pneumoniae affects virulence in a mouse model. AB - The LuxS protein is required for the biosynthesis of the type 2 autoinducer (AI 2), which is involved in quorum sensing in a wide range of bacterial species. We have determined the effects of a defined luxS mutation on the virulence of Streptococcus pneumoniae. Although the luxS mutant displayed reduced virulence relative to its wild-type parent, the type 2 strain D39, it was by no means avirulent in a mouse model. After intranasal administration, the luxS mutant was able to colonize the nasopharynx of the mouse as efficiently as the wild type. However, it was less able to spread from the nasopharynx to the lungs or the blood. Intraperitoneal coadministration studies indicated that the luxS mutant was less fit and was readily outcompeted by wild-type D39. However, when administered on its own by this route, the mutant was able to proliferate and cause fatal systemic disease, albeit at a lower rate than the wild type. Western blot analysis of whole-cell lysates of the mutant and its parent did not reveal any differences in the levels of several well-characterized virulence proteins. However, analysis of Coomassie blue-stained protein profiles after separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that mutation of luxS had pleiotropic effects on protein expression in all cellular compartments. This is consistent with the product of luxS having a regulatory role in S. pneumoniae. This is the first report of a direct role for luxS (and by inference, AI-2) in the virulence of a gram-positive pathogen. However, the fact that mutagenesis of luxS does not completely attenuate S. pneumoniae has implications for the possible use of AI-2 antagonists for treatment of pneumococcal infections. PMID- 12761101 TI - Purification and characterization of enterotoxigenic El Tor-like hemolysin produced by Vibrio fluvialis. AB - The halophilic bacterium Vibrio fluvialis is an enteric pathogen that produces an extracellular hemolysin. This hemolysin was purified to homogeneity by using sequential hydrophobic-interaction chromatography with phenyl-Sepharose CL-4B and gel filtration with Sephacryl S-200. It has a molecular weight of 63,000 and an isoelectric point of 4.6, and its hemolytic activity is sensitive to heat, proteases, and preincubation with zinc ions. The hemolysin lyses erythrocytes of the eight different animal species that we tested, is cytotoxic against Chinese hamster ovary cells in tissue culture, and elicits fluid accumulation in suckling mice. Lysis of erythrocytes occurs by a temperature-dependent binding step followed by a temperature- and pH-dependent lytic step. Fourteen of the first 20 N-terminal amino acid residues (Val-Ser-Gly-Gly-Glu-Ala-Asn-Thr-Leu-Pro-His-Val Ala-Phe-Tyr-Ile-Asn-Val-Asn-Arg) are identical to those of the El Tor hemolysin of Vibrio cholerae and the heat-labile hemolysin of Vibrio mimicus. This homology was further confirmed by PCR analysis using a 5' primer derived from the amino terminal sequence of the hemolysin and a 3' primer derived from the El Tor hemolysin structural gene. The hemolysin also reacts with antibodies to the El Tor-like hemolysin of non-O1 V. cholerae. PMID- 12761102 TI - Lipopolysaccharide-mimetic activities of a Toll-like receptor 2-stimulatory substance(s) in enterobacterial lipopolysaccharide preparations. AB - Lipopolysaccharide (LPS) preparations are known to often contain substances which activate cells through Toll-like receptor 2 (TLR2), and it is suspected that bacterial lipoproteins are responsible for this activation. We compared the mode of action of the TLR2-stimulatory substances with that of a synthetic bacterial lipopeptide (tripalmitoyl-Cys-Ser-Ser-Asn-Ala [Pam(3)CSSNA]), as well as with that of peptidoglycan. Six out of eight LPS preparations tested induced NF-kappaB dependent reporter activity in 293 cells expressing CD14 and TLR2. Phenol extract (PEX) prepared from Escherichia coli LPS by modified phenol extraction induced reporter activity in 293 cells expressing TLR2, and this activity was enhanced by coexpression of CD14, whereas the activity of Pam(3)CSSNA was not dependent on CD14. The activity of PEX, but not that of Pam(3)CSSNA or peptidoglycan, was also enhanced by LPS binding protein or serum and blocked by polymyxin B. In addition, the activity of PEX was inhibited by a lipid A precursor (compound 406) in 293 cells expressing CD14 and TLR2. These results indicate that E. coli LPS preparations contain LPS-mimetic TLR2-stimulatory substances which differ from bacterial lipopeptides or peptidoglycan. PMID- 12761103 TI - The secreted aspartyl proteinases Sap1 and Sap2 cause tissue damage in an in vitro model of vaginal candidiasis based on reconstituted human vaginal epithelium. AB - Secreted aspartyl proteinases (Saps) contribute to the ability of Candida albicans to cause mucosal and disseminated infections. A model of vaginal candidiasis based on reconstituted human vaginal epithelium (RHVE) was used to study the expression and role of these C. albicans proteinases during infection and tissue damage of vaginal epithelium. Colonization of the RHVE by C. albicans SC5314 did not cause any visible epithelial damage 6 h after inoculation, although expression of SAP2, SAP9, and SAP10 was detected by reverse transcriptase PCR. However, significant epithelial damage was observed after 12 h, concomitant with the additional expression of SAP1, SAP4, and SAP5. Additional transcripts of SAP6 and SAP7 were detected at a later stage of the artificial infection (24 h). Similar SAP expression profiles were observed in three samples isolated from human patients with vaginal candidiasis. In experimental infection, secretion of antigens Sap1 to Sap6 by C. albicans was confirmed at the ultrastructural level by using polyclonal antisera raised against Sap1 to Sap6. Addition of the aspartyl proteinase inhibitors pepstatin A and the human immunodeficiency virus proteinase inhibitors ritonavir and amprenavir strongly reduced the tissue damage of the vaginal epithelia by C. albicans cells. Furthermore, SAP null mutants lacking either SAP1 or SAP2 had a drastically reduced potential to cause tissue damage even though SAP3, SAP4, and SAP7 were up regulated in these mutants. In contrast the vaginopathic potential of mutants lacking SAP3 or SAP4 to SAP6 was not reduced compared to wild-type cells. These data provide further evidence for a crucial role of Sap1 and Sap2 in C. albicans vaginal infections. PMID- 12761104 TI - Protection against Shiga toxin 1 challenge by immunization of mice with purified mutant Shiga toxin 1. AB - Shiga toxin 1 (Stx1) of enterohemorrhagic Escherichia coli O157:H7 was cloned, and four mutant Stx1s were constructed by site-directed mutagenesis with PCR. The wild-type and mutant Stx1s with amino acid replacements at positions 167 and 170 of the A subunit were purified by one-step affinity chromatography with commercially available Globotriose Fractogel, and the mutant Stxs were used for the immunization of mice. The mutant toxins were nontoxic to Vero cells in vitro and to mice in vivo and induced the immunoglobulin G antibody against the wild type Stx1, which neutralized the cytotoxicity of Stx1. The induced antibody titers depended on the mutation at position 170 of the A subunit. The mice immunized with the mutant Stx1s were protected against a challenge of approximately 100 times the 50% lethal dose of the wild-type Stx1, suggesting that the mutant toxins are good candidates for toxoid vaccines for infection by Stx1-producing E. coli. PMID- 12761105 TI - Antigenic organization of the N-terminal part of the polymorphic outer membrane proteins 90, 91A, and 91B of Chlamydophila abortus. AB - A series of overlapping recombinant antigens, 61 to 74 residues in length, representing polymorphic outer membrane protein 90 (POMP90) of Chlamydophila abortus and two recombinant peptides spanning gene fragment p91Bf99 of POMP91B were assessed by immunoblotting to determine the antigen-binding sites of 20 monoclonal antibodies to POMP90, -91A, and -91B. The epitopes were further restricted by scanning 52 overlapping synthetic 12-mer peptides representing the N-terminal part of POMP90, and the 12-mer epitopes were then analyzed by using hexapeptides to the resolution of a single amino acid. Ten epitopes were defined: 1, TSEEFQVKETSSGT; 2, SGAIYTCEGNVCISYAGKDSPL; 3, SLVFHKNCSTAE; 4, AIYADKLTIVSGGPTLFS; 5, SPKGGAISIKDS; 6, ITFDGNKIIKTS; 7, LRAKDGFGIFFY; 7a, DGFGIF; 7b, GIFFYD; 8, IFFYDPITGGGS; 8a, FFYDPIT; 9, GKIVFSGE; and 10, DLGTTL. The 20-mer peptide LRAKDGFGIFFYDPITGGGS was a major epitope that was recognized by seven antibodies. Epitopes 7 to 10 were conserved in reference strains of the former species C. psittaci, whereas the strong antigenic peptides FYDPIT and IVFSGE were conserved among members of the genus CHLAMYDOPHILA: Epitopes 3 to 8 were located within the best-scoring beta-helical wrap (residues 148 to 293) predicted for POMP91B by the program BETAWRAP. Other studies have suggested an association of the POMPs with type V secretory autotransporter proteins. The results presented in this study provide some evidence for a passenger domain that is folded as a beta-helix pyramid with compact antigenic organization. PMID- 12761106 TI - Killing of Candida albicans by human salivary histatin 5 is modulated, but not determined, by the potassium channel TOK1. AB - Salivary histatin 5 (Hst 5), a potent toxin for the human fungal pathogen Candida albicans, induces noncytolytic efflux of cellular ATP, potassium, and magnesium in the absence of cytolysis, implicating these ion movements in the toxin's fungicidal activity. Hst 5 action on Candida resembles, in many respects, the action of the K1 killer toxin on Saccharomyces cerevisiae, and in that system the yeast plasma membrane potassium channel, Tok1p, has recently been reported to be a primary target of toxin action. The question of whether the Candida homologue of Saccharomyces Tok1p might be a primary target of Hst 5 action has now been investigated by disruption of the C. albicans TOK1 gene. The resultant strains (TOK1/tok1) and (tok1/tok1) were compared with wild-type Candida (TOK1/TOK1) for relative ATP leakage and killing in response to Hst 5. Patch-clamp measurements on Candida protoplasts were used to verify the functional deletion of Tok1p and to provide its first description in Candida. Tok1p is an outwardly rectifying, noisily gated, 40-pS channel, very similar to that described in Saccharomyces. Knockout of CaTOK1 (tok1/tok1) completely abolishes the currents and gating events characteristic of Tok1p. Also, knockout (tok1/tok1) increases residual viability of Candida after Hst 5 treatment to 27%, from 7% in the wild type, while the single allele deletion (TOK1/tok1) increases viability to 18%. Comparable results were obtained for Hst-induced ATP efflux, but quantitative features of ATP loss suggest that wild-type TOK1 genes function cooperatively. Overall, very substantial killing and ATP efflux are produced by Hst 5 treatment after complete knockout of wild-type TOK1, making clear that Tok1p channels are not the primary site of Hst 5 action, even though they do play a modulating role. PMID- 12761108 TI - Neospora caninum-infected cattle develop parasite-specific CD4+ cytotoxic T lymphocytes. AB - Cattle infected with Neospora caninum readily experience transplacental parasite transmission, presumably after maternal parasitemia, leading to abortion or birth of congenitally infected calves. Cytotoxic T lymphocytes (CTL) are important mediators of protective immunity against Toxoplasma gondii, an intracellular apicomplexan protozoan closely related to N. caninum. In this study, N. caninum specific CTL expanded from peripheral blood mononuclear cells of two major histocompatibility complex-mismatched, experimentally infected cattle were identified by using a (51)Cr release cytotoxicity assay. Enrichment and blocking of CD4(+)- and CD8(+)-T-lymphocyte effector subsets indicated that CD4(+) CTL killed N. caninum-infected, autologous target cells and that killing was mediated through a perforin/granzyme pathway. Detection and characterization of CTL responses to N. caninum in the natural, outbred, bovine host will facilitate identification of immunogens and design of immunization strategies to induce parasite-specific CTL against transplacental N. caninum transmission in cattle. PMID- 12761107 TI - Characterization of Brucella abortus O-polysaccharide and core lipopolysaccharide mutants and demonstration that a complete core is required for rough vaccines to be efficient against Brucella abortus and Brucella ovis in the mouse model. AB - Brucella abortus rough lipopolysaccharide (LPS) mutants were obtained by transposon insertion into two wbk genes (wbkA [putative glycosyltransferase; formerly rfbU] and per [perosamine synthetase]), into manB (pmm [phosphomannomutase; formerly rfbK]), and into an unassigned gene. Consistent with gene-predicted roles, electrophoretic analysis, 2-keto-3-manno-D octulosonate measurements, and immunoblots with monoclonal antibodies to O polysaccharide, outer and inner core epitopes showed no O-polysaccharide expression and no LPS core defects in the wbk mutants. The rough LPS of manB mutant lacked the outer core epitope and the gene was designated manB(core) to distinguish it from the wbk manB(O-Ag). The fourth gene (provisionally designated wa**) coded for a putative glycosyltransferase involved in inner core synthesis, but the mutant kept the outer core epitope. Differences in phage and polymyxin sensitivity, exposure or expression of outer membrane protein, core and lipid A epitopes, and lipid A acylation demonstrated that small changes in LPS core caused significant differences in B. abortus outer membrane topology. In mice, the mutants showed different degrees of attenuation and induced antibodies to rough LPS and outer membrane proteins. Core-defective mutants and strain RB51 were ineffective vaccines against B. abortus in mice. The mutants per and wbkA induced protection but less than the standard smooth vaccine S19, and controls suggested that anti O-polysaccharide antibodies accounted largely for the difference. Whereas no core-defective mutant was effective against B. ovis, S19, RB51, and the wbkA and per mutants afforded similar levels of protection. These results suggest that rough Brucella vaccines should carry a complete core for maximal effectiveness. PMID- 12761109 TI - Distribution and kinetics of lipoprotein-bound lipoteichoic acid. AB - Lipoteichoic acid (LTA), a major cell wall component of gram-positive bacteria, is an amphipathic anionic glycolipid with structural similarities to lipopolysaccharide (LPS) from gram-negative bacteria. LTA has been implicated as one of the primary immunostimulatory components that may trigger the systemic inflammatory response syndrome. Plasma lipoproteins have been shown to sequester LPS, which results in attenuation of the host response to infection, but little is known about the LTA binding characteristics of plasma lipid particles. In this study, we have examined the LTA binding capacities and association kinetics of the major lipoprotein classes under simulated physiological conditions in human whole blood (ex vivo) by using biologically active, fluorescently labeled LTA and high-performance gel permeation chromatography. The average distribution of an LTA preparation from Staphylococcus aureus in whole blood from 10 human volunteers revealed that >95% of the LTA was associated with total plasma lipoproteins in the following proportions: high-density lipoprotein (HDL), 68% +/ 10%; low-density lipoprotein (LDL), 28% +/- 8%; and very low density lipoprotein (VLDL), 4% +/- 5%. The saturation capacity of lipoproteins for LTA was in excess of 150 micro g/ml. The LTA distribution was temperature dependent, with an optimal binding between 22 and 37 degrees C. The binding of LTA by lipoproteins was essentially complete within 10 min and was followed by a subsequent redistribution from HDL and VLDL to LDL. We conclude that HDL has the highest binding capacity for LTA and propose that the loading and redistribution of LTA among plasma lipoproteins is a specific process that closely resembles that previously described for LPS (J. H. M. Levels, P. R. Abraham, A. van den Ende, and S. J. H. van Deventer, Infect. Immun. 68:2821-2828, 2001). PMID- 12761110 TI - Characterization of an iroBCDEN gene cluster on a transmissible plasmid of uropathogenic Escherichia coli: evidence for horizontal transfer of a chromosomal virulence factor. AB - The chromosomal iroBCDEN gene cluster first described for Salmonella enterica is involved in the uptake of catecholate-type siderophore compounds. An orthologous gene cluster has recently been detected in Escherichia coli strains which cause extraintestinal disease. This E. coli iroBCDEN gene cluster has an impact on virulence and has been reported to be located in a pathogenicity island on the chromosome. In this study we characterized an iro gene cluster of a uropathogenic E. coli isolate which is located on a transmissible plasmid related to the R64 plasmid of S. enterica. This cluster is highly homologous to the chromosomal iro cluster of E. coli. When introduced into an E. coli fepA cir fiu aroB mutant, IroN, but not IroBCDE, mediated the utilization of structurally related catecholate siderophores, including 2,3-dihydroxybenzoyl-L-serine, 2,3 dihydroxybenzoyl-D-ornithine, 2,3-dihydroxybenzoic acid, and enterochelin. This study supports the idea of an ongoing horizontal transfer of putative virulence factors and the mobilization of single virulence gene clusters, which lead to a modular assembly of virulence determinants such as pathogenicity islands. PMID- 12761111 TI - Mutational analysis of the enzymatic domain of Clostridium difficile toxin B reveals novel inhibitors of the wild-type toxin. AB - Toxin B (TcdB), a major Clostridium difficile virulence factor, glucosylates and inactivates the small GTP-binding proteins Rho, Rac, and Cdc42. In the present study we provide evidence that enzymatically inactive fragments of the TcdB enzymatic domain are effective intracellular inhibitors of native TcdB. Site directed and deletion mutants of the TcdB enzymatic region (residues 1 to 556), lacking receptor binding and cell entry domains, were analyzed for attenuation of glucosyltransferase and glucosylhydrolase activity. Five of six derivatives from TcdB(1-556) were found to be devoid of enzymatic activity. In order to facilitate cell entry, mutants were genetically fused to lfn, which encodes the protective antigen binding region of anthrax toxin lethal factor and mediates the cell entry of heterologous proteins. In line with reduced enzymatic activity, the mutants also lacked cytotoxicity. Remarkably, pretreatment or cotreatment of cells with four of the mutants provided protection against the cytotoxic effects of native TcdB. Furthermore, a CHO cell line expressing enzymatically active TcdB(1-556) was also protected by the mutant-derived inhibitors, suggesting that inhibition occurred at an intracellular location. Protection also was afforded by the inhibitor to cells treated with Clostridium sordellii lethal toxin (TcsL), which uses the same cosubstrate as TcdB but shares Rac only as a common substrate target. Finally, the inhibitor did not provide protection against Clostridium novyi alpha-toxin (Tcnalpha), which shares similar substrates with TcdB yet uses a different cosubstrate. This is the first report to demonstrate that the potential exists to inhibit toxins at their intracellular site of action by using inactive mutants. PMID- 12761112 TI - The immunoglobulin D-binding protein MID from Moraxella catarrhalis is also an adhesin. AB - The Moraxella catarrhalis immunoglobulin D (IgD)-binding protein (MID) is a 200 kDa outer membrane protein displaying a unique and specific affinity for human IgD. MID is found in the majority of M. catarrhalis strains. In the present paper, we show that MID-expressing M. catarrhalis strains agglutinate human erythrocytes and bind to type II alveolar epithelial cells. In contrast, M. catarrhalis isolates with low MID expression levels and two mutants deficient in MID, but with readily detectable UspA1 expression, do not agglutinate erythrocytes and have a 50% lower adhesive capacity. To examine the adhesive part of MID, the protein was dissected into nine fragments covering the entire molecule. The truncated MID proteins were expressed in Escherichia coli, purified, and used for raising polyclonal antibodies in rabbits. Interestingly, by using recombinant fragments, we show that the hemagglutinating and adhesive part of MID is localized within the 150-amino-acid fragment MID(764-913). In addition, antibodies against full-length MID, MID(764-913), or a 30-amino-acid consensus sequence (MID(775-804)) inhibited adhesion to alveolar epithelial cells. Antibodies against UspA1, an outer membrane protein expressed in essentially all M. catarrhalis strains, also inhibited adhesion, suggesting that both MID and UspA1 are needed for optimal attachment to epithelial cells. Taken together, in addition to MID-dependent IgD binding, we have demonstrated that the outer membrane protein MID is a novel adhesin that would be a suitable target for a future vaccine against M. catarrhalis. PMID- 12761114 TI - Association of Actinobacillus pleuropneumoniae capsular polysaccharide with virulence in pigs. AB - The capsular polysaccharide (CP) of Actinobacillus pleuropneumoniae is required for virulence of the bacteria in swine. However, a molecular investigation of whether the type or quantity of CP affects A. pleuropneumoniae virulence has not been reported. To initiate this investigation, a DNA region downstream of conserved genes required for CP export in A. pleuropneumoniae serotype 1 was cloned and sequenced. Three open reading frames, designated cps1A, cps1B, and cps1C, were identified that had amino acid homology to bacterial carbohydrate biosynthesis genes. A kanamycin resistance cassette (Kan(r)) was inserted into a 750-bp deletion spanning cps1AB or into a 512-bp deletion in cps1B only, and the constructs were cloned in a suicide vector. The Kan(r) gene was then transferred into the chromosome of strain 4074 by homologous recombination to produce strain 4074Deltacps1N and strain 4074Deltacps1B, respectively. Strain 4074Deltacps1N produced no detectable CP, but strain 4074Deltacps1B made 15% of the serotype 1 CP made by the parent strain, 4074, as determined by enzyme-linked immunosorbent assay and precipitation of free CP. The cps1ABC genes of strain 4074 and the cps5ABC and cps5ABCDE genes of serotype 5a strain J45 were cloned into the shuttle vector pLS88 and electroporated into 4074Deltacps1N to produce 4074Deltacps1N(pABcps101), 4074Deltacps1N(pJMLcps53), and 4074Deltacps1N(pABcps55), respectively. Strain 4074Deltacps1N(pABcps101) produced about 33% of the serotype 1 CP produced by strain 4074. Strains 4074Deltacps1N(pJMLcps53) and 4074Deltacps1N(pABcps55) produced serotype 5a CP in similar quantity or in fourfold excess, respectively, to that produced by strain 4074. With intratracheal challenge in pigs at similar dosages, the order of virulence of strains producing serotype 1 CP (assessed by mortality, lung consolidation, hemorrhage, and fibrinous pleuritis) was the following: strain 4074 > strain 4074Deltacps1N(pABcps101) > or = strain 4074Deltacps1N > strain 4074Deltacps1B. Strain 4074Deltacps1N(pJMLcps53) was less virulent than strain 4074Deltacps1N(pABcps55). However, both strains produced serotype 5a CP in similar or greater quantities than was observed for production of serotype 1 CP by the parent strain, 4074, but were less virulent than the parent strain. Therefore, the amount of serotype 1 or 5a CP produced by isogenic strains of A. pleuropneumoniae correlated with the virulence of the bacteria in pigs. However, virulence was also influenced by the type of CP produced or by its mechanism of expression. PMID- 12761113 TI - Secretin of the enteropathogenic Escherichia coli type III secretion system requires components of the type III apparatus for assembly and localization. AB - At least 16 proteins are thought to be involved in forming the enteropathogenic Escherichia coli (EPEC) type III translocation apparatus which delivers virulence factors into host cells, yet their function and location have not been determined. A biochemical analysis was performed on three components: EscN, a predicted cytoplasmic ATPase; EscV, a predicted inner membrane protein; and EscC, a predicted outer membrane secretin. Wild-type EPEC and mutants constructed in these genes were fractionated by lysozyme treatment, ultracentrifugation, and selective detergent extraction. Fractionation revealed that the type III effectors Tir and EspB required a complete type III apparatus for any degree of export by EPEC, suggesting a continuous channel. Epitope-tagged EscC, EscV, and EscN were localized by fractionation, confirming computer modeling predictions for their location. Transcomplementation experiments revealed that localization of EscV and EscN was unaffected by mutations in other examined type III components. Remarkably, localization of EscC was altered in escV or escN mutants, where EscC accumulated in the periplasm. EscC was correctly localized in the escF needle component mutant, indicating that secretin localization is independent of needle formation. These data indicate that, contrary to previous indications, correct insertion and function of EscC secretin in the outer membrane depends not only on the sec-dependent secretion pathway but also on other type III apparatus components. PMID- 12761115 TI - Study of capsular polysaccharide from Vibrio parahaemolyticus. AB - The leading cause of food poisoning in both Taiwan and Japan is Vibrio parahaemolyticus infection, whose mechanism of enteropathogenesis is still unclear. To evaluate whether surface components are responsible for the intestinal adhesion of V. parahaemolyticus, we have developed a novel method for isolating the capsular polysaccharide (CPS) from V. parahaemolyticus (serotype O4:K8). We found that culturing of V. parahaemolyticus in broth for 1 week or more changed the colony form of the bacteria on an agar plate from opaque to translucent. The translucent colonies of V. parahaemolyticus contained little CPS and exhibited a much lower level of adherence to epithelial cells (Int-407) than the opaque colonies of the bacteria. Incubation of V. parahaemolyticus in medium supplemented with bile increased the levels of CPS and adherence. Treatment of V. parahaemolyticus with anti-CPS but not anti-LPS serum decreased the level of bacterial adherence. In addition, purified CPS bound to epithelial cells in a dose-dependent manner. Intranasal administration of CPS to mice in the presence of adjuvants such as immunostimulatory sequence oligodeoxynucleotides or cholera toxin elicited CPS-specific mucosal and systemic immune responses. These results indicate that CPS plays an important role in the adherence of V. parahaemolyticus to its target cells and may be considered a potential target for the development of a vaccine against this pathogen. PMID- 12761117 TI - A mosaic pathogenicity island made up of the locus of enterocyte effacement and a pathogenicity island of Escherichia coli O157:H7 is frequently present in attaching and effacing E. coli. AB - Enteropathogenic Escherichia coli (EPEC) and enterohemorragic E. coli (EHEC) possess a pathogenicity island (PAI), termed the locus of enterocyte effacement (LEE), which confers the capability to cause the characteristic attaching and effacing lesions of the brush border. Due to this common property, these organisms are also termed attaching and effacing E. coli (AEEC). Sequencing of the EHEC O157 genome recently revealed the presence of other putative PAIs in the chromosome of this organism. In this article, we report on the presence of four of those PAIs in a panel of 133 E. coli strains belonging to different pathogroups and serotypes. One of these PAIs, termed O122 in strain EDL 933 and SpLE3 in strain Sakai, was observed in most of the AEEC strains examined but not in the other groups of E. coli. It was also found to contain the virulence associated gene efa1/lifA. In EHEC O157, PAI O122 is located 0.7 Mb away from the LEE. Conversely, we demonstrated that in many EHEC non-O157 strains and EPEC strains belonging to eight serogroups, PAI O122 and the LEE are physically linked to form a cointegrated structure. This structure can be considered a mosaic PAI that could have been acquired originally by AEEC. In some clones, such as EHEC O157, the LEE-O122 mosaic PAI might have undergone recombinational events, resulting in the insertion of the portion referred to as PAI O122 in a different location. PMID- 12761116 TI - Differential induction of interleukin-10 and interleukin-12 in dendritic cells by microbial toll-like receptor activators and skewing of T-cell cytokine profiles. AB - Dendritic cells (DCs) discriminate different microbial pathogens and induce T cell responses of appropriate effector phenotypes accordingly. Microbial recognition and differentiation are mediated in part by pattern recognition receptors such as Toll-like receptors (TLRs), whereas the development of T-cell effector functions is critically dependent on DC-derived cytokines such as interleukin-12 (IL-12) and IL-10. However, it is not entirely clear to what extent various microbial TLR activators could induce different functional states of DCs that favor different T-cell effector phenotypes. Toward a better understanding of this issue, we examined IL-10 and IL-12 production and T-cell polarizing potentials of murine bone marrow-derived DCs after stimulation by three microbial TLR activators, namely, lipopolysaccharide (LPS), peptidoglycan (PGN), and zymosan. We found that the three stimuli induced drastically different profiles of IL-10 and IL-12 production in DCs. Further, these stimuli differentially conditioned CD40-dependent IL-10 and IL-12 production by DCs. Finally, LPS-, PGN-, and zymosan-stimulated DCs primed distinct T-cell cytokine profiles. Our results support the notion that microbe-specific information sensed through different TLRs by DCs is linked to differential Th priming through DC derived cytokines. PMID- 12761118 TI - Structural and functional consequences of cleavage of human secretory and human serum immunoglobulin A1 by proteinases from Proteus mirabilis and Neisseria meningitidis. AB - The cleavage of human serum monomeric immunoglobulin A1 (IgA1) and human secretory IgA1 (S-IgA1) by IgA1 proteinase of Neisseria meningitidis and cleavage by the proteinase from Proteus mirabilis have been compared. For serum IgA1, both proteinases cleaved only the alpha chain. N. meningitidis proteinase cleaved only in the hinge. P. mirabilis proteinase sequentially removed the tailpiece, the CH3 domain, and the CH2 domain. The cleavage of S-IgA1 by N. meningitidis proteinase occurred only in the hinge and was as rapid as that of serum IgA1. P. mirabilis proteinase predominantly cleaved the secretory component (SC) of S-IgA1. The SC of S-IgA1, whether cleaved or not, appeared to protect the alpha1 chain. Purified Fc fragment derived from the cleavage of serum IgA1 by N. meningitidis proteinase stimulated a respiratory burst in neutrophils through Fcalpha receptors, whereas the (Fcalpha1)(2)-SC fragment from digested S-IgA1 did not. The loss of the tailpiece from serum IgA1 treated with P. mirabilis proteinase had little effect, but the loss of the CH3 domain was concurrent with a rapid loss in the ability to bind to Fcalpha receptors. S-IgA1 treated with P. mirabilis proteinase under the same conditions retained the ability to bind to Fcalpha receptors. The results are consistent with the Fcalpha receptor binding site being at the CH2-CH3 interface. These data shed further light on the structure of S-IgA1 and indicate that the binding site for the Fcalpha receptor in S-IgA is protected by SC, thus prolonging its ability to activate phagocytic cells at the mucosal surface. PMID- 12761119 TI - Helicobacter pylori infection induces interleukin-8 receptor expression in the human gastric epithelium. AB - The gastric pathogen Helicobacter pylori is known to activate multiple proinflammatory signaling pathways in epithelial cells. In this study, we addressed the question of whether expression of the interleukin-8 receptors IL 8RA (CXCR1) and IL-8RB (CXCR2) is upregulated in H. pylori-infected human gastric biopsy samples. Biopsy samples from patients infected with H. pylori strains harboring the cag pathogenicity island (PAI) expressed larger amounts of both receptors. In addition, IL-8RB expression was induced in the gastric epithelial cell line AGS upon infection with a clinical isolate containing the cag PAI, while a strain lacking the cag PAI did not. Our finding suggests that gastric epithelial cells express IL-8R in response to H. pylori infection. PMID- 12761120 TI - c-Jun NH2-terminal kinase-mediated signaling is essential for Pseudomonas aeruginosa ExoS-induced apoptosis. AB - As an opportunistic bacterial pathogen, Pseudomonas aeruginosa mainly affects immunocompromised individuals as well as patients with cystic fibrosis. In a previous study, we showed that ExoS of P. aeruginosa, when injected into host cells through a type III secretion apparatus, functions as an effector molecule to trigger apoptosis in various tissue culture cells. Here, we show that injection of the ExoS into HeLa cells activates c-Jun NH(2)-terminal kinase (JNK) phosphorylation while shutting down ERK1/2 and p38 phosphorylation. Inhibiting JNK activation by expression of a dominant negative JNK1 or with a specific JNK inhibitor abolishes ExoS-triggered apoptosis, demonstrating the requirement for JNK-mediated signaling. Following JNK phosphorylation, cytochrome c is released into the cytosol, leading to the activation of caspase 9 and eventually caspase 3. Although c-Jun phosphorylation is also observed as a result of JNK activation, ongoing host protein synthesis is not essential for the apoptotic induction, suggesting that c-Jun- or other AP-1-driven activation of gene expression is dispensable in this process. Therefore, ExoS has opposing effects on different cellular pathways that regulate apoptosis: it shuts down host cell survival signal pathways by inhibiting ERK1/2 and p38 activation, and it activates proapoptotic pathways through activation of JNK1/2 leading ultimately to cytochrome c release and activation of caspases. These results highlight the modulation of host cell signaling by the type III secretion system during interaction between P. aeruginosa and host cells. PMID- 12761121 TI - Global analysis of Borrelia burgdorferi genes regulated by mammalian host specific signals. AB - Lyme disease is a tick-borne infection that can lead to chronic, debilitating problems if not recognized or treated appropriately. Borrelia burgdorferi, the causative agent of Lyme disease, is maintained in nature by a complex enzootic cycle involving Ixodes ticks and mammalian hosts. Many previous studies support the notion that B. burgdorferi differentially expresses numerous genes and proteins to help it adapt to growth in the mammalian host. In this regard, several studies have utilized a dialysis membrane chamber (DMC) cultivation system to generate "mammalian host-adapted" spirochetes for the identification of genes selectively expressed during mammalian infection. Here, we have exploited the DMC cultivation system in conjunction with microarray technology to examine the global changes in gene expression that occur in the mammalian host. To identify genes regulated by only mammal-specific signals and not by temperature, borrelial microarrays were hybridized with cDNA generated either from organisms temperature shifted in vitro from 23 degrees C to 37 degrees C or from organisms cultivated by using the DMC model system. Statistical analyses of the combined data sets revealed that 125 genes were expressed at significantly different levels in the mammalian host, with almost equivalent numbers of genes being up- or down-regulated by B. burgdorferi within DMCs compared to those undergoing temperature shift. Interestingly, during DMC cultivation, the vast majority of genes identified on the plasmids were down-regulated (79%), while the differentially expressed chromosomal genes were almost entirely up-regulated (93%). Global analysis of the upstream promoter regions of differentially expressed genes revealed that several share a common motif that may be important in transcriptional regulation during mammalian infection. Among genes with known or putative functions, the cell envelope category, which includes outer membrane proteins, was found to contain the most differentially expressed genes. The combined findings have generated a subset of genes that can now be further characterized to help define their role or roles with regard to B. burgdorferi virulence and Lyme disease pathogenesis. PMID- 12761122 TI - Prior exposure to live Mycobacterium bovis BCG decreases Cryptococcus neoformans induced lung eosinophilia in a gamma interferon-dependent manner. AB - Some common childhood infections appear to prevent the development of atopy and asthma. In some Mycobacterium bovis BCG-vaccinated populations, strong delayed type hypersensitivity responses to mycobacterial antigens are associated with a reduced risk of atopy. Although BCG exposure decreases allergen-induced lung eosinophilia in animal models, little attention has been given to the effect of immunity to BCG on responses against live pathogens. We used the murine Cryptococcus neoformans infection model to investigate whether prior BCG infection can alter such responses. The present study shows that persistent pulmonary BCG infection of C57BL/6 mice induced an increase in gamma interferon, a reduction in interleukin-5, and a decrease in lung eosinophilia during subsequent Cryptococcus infection. This effect was long lasting, depended on the presence of live bacteria, and required persistence of mycobacterial infection in the lung. Reduction of eosinophilia was less prominent after infection with a mutant BCG strain (DeltahspR), which was rapidly cleared from the lungs. These observations have important implications for the development of vaccines designed to prevent Th2-mediated disease and indicate that prior lung BCG vaccination can alter the pattern of subsequent host inflammation. PMID- 12761123 TI - Contribution of proton-translocating proteins to the virulence of Salmonella enterica serovars Typhimurium, Gallinarum, and Dublin in chickens and mice. AB - We investigated the attenuating effects of a range of respiratory chain mutations in three Salmonella serovars which might be used in the development of live vaccines. We tested mutations in nuoG, cydA, cyoA, atpB, and atpH in three serovars of Salmonella enterica: Typhimurium, Dublin, and Gallinarum. All three serovars were assessed for attenuation in their relevant virulence assays of typhoid-like infections. Serovar Typhimurium was assessed in 1-day-old chickens and the mouse. Serovar Gallinarum 9 was assessed in 3-week-old chickens, and serovar Dublin was assessed in 6-week-old mice. Our data show variation in attenuation for the nuoG, cydA, and cyoA mutations within the different serovar host combinations. However, mutations in atpB and atpH were highly attenuating for all three serovars in the various virulence assays. Further investigation of the mutations in the atp operon showed that the bacteria were less invasive in vivo, showing reduced in vitro survival within phagocytic cells and reduced acid tolerance. We present data showing that this reduced acid tolerance is due to an inability to adapt to conditions rather than a general sensitivity to reduced pH. The data support the targeting of respiratory components for the production of live vaccines and suggest that mutations in the atp operon provide suitable candidates for broad-spectrum attenuation of a range of Salmonella serovars. PMID- 12761124 TI - Disparity in functional activity between serum anticapsular antibodies induced in adults by immunization with an investigational group A and C Neisseria meningitidis-diphtheria toxoid conjugate vaccine and by a polysaccharide vaccine. AB - Polysaccharide-protein conjugate vaccines elicit higher concentrations of serum anticapsular antibody in infants and children than do unconjugated polysaccharide vaccines. The conjugate-induced antibodies also have higher avidity and complement-mediated bactericidal activity. Similar vaccine-related differences in the magnitude or functional activity of antibody are observed infrequently in immunized adults. We compared the antibody responses of adults immunized with an investigational group A and C meningococcal conjugate vaccine to those elicited by an unconjugated meningococcal polysaccharide vaccine. Although there were no significant differences between the respective geometric mean bactericidal titers of the two vaccine groups, it took, on average, three- to fourfold higher concentrations of polysaccharide-induced serum anticapsular antibody to achieve 50% complement-mediated bacteriolysis than conjugate-induced antibody (P < 0.001 for groups A and C). At limiting doses, the polysaccharide-induced anticapsular antibodies also were less effective in conferring passive protection against meningococcal bacteremia in infant rats challenged with a group C strain (P < 0.04). The avidity index of the group C antibodies was higher in the conjugate vaccine group than in the polysaccharide vaccine group (P < 0.005). The disparities in the functional activity of the anticapsular antibodies elicited in adults by the two vaccines imply fundamental differences in the respective B-cell populations stimulated. PMID- 12761125 TI - Immunity profiles of wild-type and recombinant shiga-like toxin-encoding bacteriophages and characterization of novel double lysogens. AB - The pathogenicity of Shiga-like toxin (stx)-producing Escherichia coli (STEC), notably serotype O157, the causative agent of hemorrhagic colitis, hemolytic uremic syndrome, and thrombotic thrombocytopenic purpura, is based partly on the presence of genes (stx(1) and/or stx(2)) that are known to be carried on temperate lambdoid bacteriophages. Stx phages were isolated from different STEC strains and found to have genome sizes in the range of 48 to 62 kb and to carry either stx(1) or stx(2) genes. Restriction fragment length polymorphism patterns and sodium dodecyl sulfate-polyacrylamide gel electrophoresis protein profiles were relatively uninformative, but the phages could be differentiated according to their immunity profiles. Furthermore, these were sufficiently sensitive to enable the identification and differentiation of two different phages, both carrying the genes for Stx2 and originating from the same STEC host strain. The immunity profiles of the different Stx phages did not conform to the model established for bacteriophage lambda, in that the pattern of individual Stx phage infection of various lysogens was neither expected nor predicted. Unexpected differences were also observed among Stx phages in their relative lytic productivity within a single host. Two antibiotic resistance markers were used to tag a recombinant phage in which the stx genes were inactivated, enabling the first reported observation of the simultaneous infection of a single host with two genetically identical Stx phages. The data demonstrate that, although Stx phages are members of the lambdoid family, their replication and infection control strategies are not necessarily identical to the archetypical bacteriophage lambda, and this could be responsible for the widespread occurrence of stx genes across a diverse range of E. coli serotypes. PMID- 12761127 TI - Experimental validation of low virulence in field strains of Listeria monocytogenes. AB - Several reports have described Listeria monocytogenes strains which were nonpathogenic or weakly pathogenic, but little is known about these low-virulence strains. We found that 9 field L. monocytogenes strains were hypovirulent and 17 were avirulent, based on the number of mice contaminated and the colonization of their spleens after subcutaneous inoculation. All these strains possessed the known virulence genes. We have now assessed the low virulence of these strains in other assays before determining how they differ from virulent strains. We have shown that the low-virulence strains exhibited a phenotypic stability and were not a mixture of virulent and avirulent bacteria. They did not recover virulence after many passages in mice and colonized the spleens of mice more poorly than virulent strains after i.v. inoculation. Their lethal capacities, determined by 50% lethal dose (LD(50)), were lower than those of virulent strains. Like Listeria innocua, 14 of 17 avirulent strains had no LD(50) and were eliminated by the lymph nodes after subcutaneous inoculation. The virulent, hypovirulent, and avirulent strains were always significantly different, whatever the tests of virulence used, confirming the importance of these low-virulence field strains in identifying the proteins involved in virulence. PMID- 12761126 TI - Antigenic composition of Borrelia burgdorferi during infection of SCID mice. AB - The general concept that during infection of mice the Borrelia burgdorferi surface protein composition differs profoundly from that of tick-borne or in vitro-cultivated spirochetes is well established. Specific knowledge concerning the differences is limited because the small numbers of spirochetes present in tissue have not been amenable to direct compositional analysis. In this report we describe novel means for studying the antigenic composition of host-adapted Borrelia (HAB). The detergent Triton X-114 was used to extract the detergent phase HAB proteins from mouse ears, ankles, knees, and hearts. Immunoblot analysis revealed a profile distinct from that of in vitro-cultivated Borrelia (IVCB). OspA and OspB were not found in the tissues of SCID mice 17 days after infection. The amounts of antigenic variation protein VlsE and the relative amounts of its transcripts were markedly increased in ear, ankle, and knee tissues but not in heart tissue. VlsE existed as isoforms having both different unit sizes and discrete lower molecular masses. The hydrophobic smaller forms of VlsE were also found in IVCB. The amounts of the surface protein (OspC) and the decorin binding protein (DbpA) were increased in ear, ankle, knee, and heart tissues, as were the relative amounts of their transcripts. Along with these findings regarding VlsE, OspC, and DbpA, two-dimensional immunoblot analysis with immune sera also revealed additional details of the antigenic composition of HAB extracted from ear, heart, and joint tissues. A variety of novel antigens, including antigens with molecular masses of 65 and 30 kDa, were found to be upregulated in mouse tissues. Extraction of hydrophobic B. burgdorferi antigens from tissue provides a powerful tool for determining the antigenic composition of HAB. PMID- 12761128 TI - Inhibition of interleukin-17 prevents the development of arthritis in vaccinated mice challenged with Borrelia burgdorferi. AB - We showed that Borrelia burgdorferi-vaccinated interferon gamma-deficient (IFN gamma(0)) mice challenged with the Lyme spirochete developed a prominent chronic severe destructive osteoarthropathy. The immune response underlying the development of the severe destructive arthritis involves interleukin-17 (IL-17). Treatment of vaccinated IFN-gamma(0) mice challenged with B. burgdorferi with anti-IL-17 antibody delayed the onset of swelling of the hind paws but, more importantly, inhibited the development of arthritis. Histopathologic examination confirmed that treatment with anti-IL-17 antibody prevented the destructive arthropathy seen in vaccinated and challenged IFN-gamma(0) mice. Similar preventive results were obtained when vaccinated and challenged IFN-gamma(0) mice were treated with anti-IL-17 receptor antibody or sequentially with anti-IL-17 antibody followed by anti-IL-17 receptor antibody. By contrast, treatment of vaccinated and challenged IFN-gamma(0) mice with recombinant IL-17 (rIL-17) did not alter the development and progression of arthritis found in vaccinated and challenged IFN-gamma(0) mice without treatment with rIL-17. Therapeutic intervention may be a realistic approach to prevent arthritis, especially if IL 17 is involved in the perpetuation of chronic or intermittent arthritis. PMID- 12761129 TI - Host susceptibility to the attaching and effacing bacterial pathogen Citrobacter rodentium. AB - Many studies have shown that genetic susceptibility plays a key role in determining whether bacterial pathogens successfully infect and cause disease in potential hosts. Surprisingly, whether host genetics influence the pathogenesis of attaching and effacing (A/E) bacteria such as enteropathogenic and enterohemorrhagic Escherichia coli has not been examined. To address this issue, we infected various mouse strains with Citrobacter rodentium, a member of the A/E pathogen family. Of the strains tested, the lipopolysaccharide (LPS) nonresponder C3H/HeJ mouse strain experienced more rapid and extensive bacterial colonization than did other strains. Moreover, the high bacterial load in these mice was associated with accelerated crypt hyperplasia, mucosal ulceration, and bleeding, together with very high mortality rates. Interestingly, the basis for the increased susceptibility was not due to LPS hyporesponsiveness, as the genetically related but LPS-responsive C3H/HeOuJ and C3H/HeN mouse strains were also susceptible to infection. Analysis of the intestinal pathology in these susceptible strains revealed significant crypt epithelial cell apoptosis (terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end label staining) as well as bacterial translocation to the mesenteric lymph nodes. Further studies with infection of SCID (T- and B-lymphocyte-deficient) C3H/HeJ mice demonstrated that loss of lymphocytes had no effect on bacterial numbers but did reduce crypt cell apoptosis and delayed mortality. These studies thus identify the adaptive immune system, crypt cell apoptosis, and bacterial translocation but not LPS responsiveness as contributing to the tissue pathology and mortality seen during C. rodentium infection of highly susceptible mouse strains. Determining the basis for these strains' susceptibility to intestinal colonization by an A/E pathogen will be the focus of future studies. PMID- 12761130 TI - Nontypeable Haemophilus influenzae gene expression induced in vivo in a chinchilla model of otitis media. AB - The gram-negative bacterium nontypeable Haemophilus influenzae (NTHI) is the predominant pathogen in chronic otitis media with effusion and, with Streptococcus pneumoniae and Moraxella catarrhalis, is a causative agent of acute otitis media. To identify potential virulence determinants, bacterial gene expression was monitored by differential fluorescence induction during early disease progression in one specific anatomical niche of a chinchilla model of NTHI-induced otitis media. Genomic DNA fragments from NTHI strain 86-028NP were cloned upstream of the promoterless gfpmut3 gene. NTHI strain 86-028NP served as the host for the promoter trap library. Pools of 2,000 transformants were inoculated into the left and right middle ear cavities of chinchillas. Middle ear effusions were recovered by epitympanic tap at 24 and 48 h, and clones containing promoter elements that were induced in vivo and producing green fluorescent protein were isolated by two-color fluorescence-activated cell sorting. Insert DNA was sequenced and compared to the complete genome sequence of H. influenzae strain Rd. In a screen of 16,000 clones, we have isolated 44 clones that contain unique gene fragments encoding biosynthetic enzymes, metabolic and regulatory proteins, and hypothetical proteins of unknown function. An additional eight clones contain gene fragments unique to our NTHI isolate. Using quantitative reverse transcription-PCR, we have confirmed that 26 clones demonstrated increased gene expression in vivo relative to expression in vitro. These data provide insight into the response of NTHI bacteria as they sense and respond to the middle ear microenvironment during early events of otitis media. PMID- 12761131 TI - Attenuated Yersinia pseudotuberculosis carrier vaccine for simultaneous antigen specific CD4 and CD8 T-cell induction. AB - Yersinia pseudotuberculosis employs a type III secretion system for targeting of several virulence factors directly to the cytosol of eukaryotic cells. This protein translocation mechanism mediates the ability of Yersinia to resist phagocytosis and is required for sustained extracellular bacterial replication. In the present study, the Yersinia outer protein E (YopE) was used as a carrier molecule for type III-dependent secretion and translocation of listeriolysin O (LLO) from Listeria monocytogenes. In comparison to wild-type Yersinia, an attenuated Y. pseudotuberculosis yopK-null mutant strain hypertranslocates chimeric YopE/LLO into the cytosol of macrophages, resulting in enhanced major histocompatibility complex (MHC) class I-restricted antigen presentation of an LLO-derived CD8 T-cell epitope. Remarkably, T-cell activation assays also revealed a superior ability of translocated over secreted LLO to induce MHC class II-restricted antigen presentation. These in vitro observations were confirmed after immunization of mice with a single dose of the yopK-null mutant strain. Animals orally inoculated with recombinant Yersinia expressing translocated chimeric YopE/LLO revealed high numbers of gamma interferon-producing LLO specific CD4 and CD8 T cells. For the first time, it is shown that cytosolic antigen display mediated by an extracellular bacterial carrier vaccine results in simultaneous CD4 and CD8 T-cell priming, conferring protection against an intracellular pathogen. PMID- 12761132 TI - Deletion of the gene encoding p60 in Listeria monocytogenes leads to abnormal cell division and loss of actin-based motility. AB - Protein p60 encoded by the iap gene is regarded as an essential gene product of Listeria monocytogenes. Here we report, however, the successful construction of a viable iap deletion mutant of L. monocytogenes EGD. The mutant, which produces no p60, shows abnormal septum formation and tends to form short filaments and hooked forms during logarithmic growth. These abnormal bacterial cells break into almost normal sized single bacteria in the late-stationary-growth phase. The iap mutant is strongly attenuated in a mouse model after intravenous injection, demonstrating the importance of p60 during infection, and the invasiveness of the Deltaiap mutant for 3T6 fibroblasts and Caco-2 epithelial cells is slightly reduced. Upon uptake by epithelial cells and macrophages, the iap mutant escapes from the phagosome into the cytosol with the same efficiency as the wild-type strain, and the mutant bacteria also grow intracellularly at a rate similar to that of the wild-type strain. Intracellular movement and cell-to-cell spread are drastically reduced in various cell lines, since the iap-negative bacteria fail to induce the formation of actin tails. However, the bacteria are covered with actin filaments. Most intracellular bacteria show a nonpolar and uneven distribution of ActA around the cell, in contrast to that for the wild-type strain, where ActA is concentrated at the old pole. In an iap(+) revertant strain that produces wild-type levels of p60, intracellular movement, cell-to-cell spread, and polar distribution of ActA are fully restored. In vitro analysis of ActA distribution on the filaments of the Deltaiap strain shows that the loss of bacterial septum formation leads to ActA accumulation at the presumed division sites. In the light of data presented here and elswhere, we propose to rename iap (invasion-associated protein) cwhA (cell wall hydrolase A). PMID- 12761133 TI - Genetic diversity and antigenic polymorphism in Plasmodium falciparum: extensive serological cross-reactivity between allelic variants of merozoite surface protein 2. AB - Diversity in the surface antigens of malaria parasites is generally assumed to be a mechanism for immune evasion, but there is little direct evidence that this leads to evasion of protective immunity. Here we show that alleles of the highly polymorphic merozoite surface protein 2 (MSP-2) can be grouped (within the known dimorphic families) into distinct serogroups; variants within a serogroup show extensive serological cross-reactivity. Cross-reactive epitopes are immunodominant, and responses to them may be boosted at the expense of responses to novel epitopes (original antigenic sin). The data imply that immune selection explains only some of the diversity in the msp-2 gene and that MSP-2 vaccines may need to include only a subset of the known variants in order to induce pan reactive antibodies. PMID- 12761134 TI - Helicobacter pylori activates Toll-like receptor 4 expression in gastrointestinal epithelial cells. AB - Helicobacter pylori activates the transcription factor NF-kappaB, leading to proinflammatory cytokine production by gastric epithelial cells. However, the receptors for the initial bacterial interaction with host cells which activate downstream signaling events have not been completely defined. Recently, it has been shown that microbial components activate Toll-like receptors (TLRs), thereby leading to AP-1- and NF-kappaB-dependent transcription and resulting in the production of proinflammatory cytokines. The aim of this study was to determine whether H. pylori activates TLR4. Reverse transcription-PCR showed that both type I and type II H. pylori clinical isolates induced TLR4 mRNA expression in AGS cells compared with that by uninfected controls. H. pylori upregulated TLR4 protein expression in two gastric epithelial cell lines (AGS and MKN45) and one intestinal epithelial cell line (T84). Monoclonal TLR4 antibody inhibited lipopolysaccharide-induced interleukin-8 secretion from THP-1 macrophages but not from gastric epithelial cells infected with H. pylori. H. pylori demonstrated increased adherence to CHO TLR4-transfected cells compared with that to both CHO TLR2-transfected and nontransfected CHO cells (P < 0.01). These results indicate that H. pylori activates TLR4 expression in epithelial cells and that TLR4 can serve as a receptor for H. pylori binding. PMID- 12761136 TI - The rovA mutant of Yersinia enterocolitica displays differential degrees of virulence depending on the route of infection. AB - Yersinia enterocolitica is an invasive enteric pathogen that causes significant inflammatory disease. Recently, we identified and characterized a global regulator of virulence (rovA). When mice are infected orally with the rovA mutant they are attenuated by 50% lethal dose (LD(50)) analysis and have altered kinetics of infection. Most significantly, mice orally infected with the rovA mutant have greatly reduced inflammation in the Peyer's patches compared to those infected with wild-type Y. enterocolitica. However, we present data here indicating that when the rovA mutant bacteria are delivered intraperitoneally (i.p.), they are significantly more virulent than when delivered orally. The i.p. LD(50) for the rovA mutant is only 10-fold higher than that of the wild-type Y. enterocolitica, and there are significant inflammatory responses to the rovA mutant that are evident in the liver and spleen. Altogether, these data suggest that the RovA regulon may be required for the early events of the infection that occur in the Peyer's patches. Furthermore, these data suggest that the RovA regulon may be dispensable for Y. enterocolitica systemic disease and inflammatory responses if the Peyer's patches are bypassed. PMID- 12761135 TI - Gamma interferon augments the intracellular pathway for lipopolysaccharide (LPS) recognition in human intestinal epithelial cells through coordinated up regulation of LPS uptake and expression of the intracellular Toll-like receptor 4 MD-2 complex. AB - Although some intestinal epithelial cell lines are known to respond to lipopolysaccharide (LPS), understanding of the relationship between LPS responsiveness and the expression of LPS receptors or factors regulating LPS responsiveness of intestinal epithelial cell lines is incomplete. In this study, we demonstrate that commonly studied human intestinal epithelial cell lines can be classified into at least three different types on the basis of LPS responsiveness, Toll-like receptor-4 (TLR4) expression, and the effects of gamma interferon (IFN-gamma) on LPS responsiveness. The first phenotype, which includes the HCT-116 and Caco-2 cell lines, is characterized by relative hyporesponsiveness to LPS and diminished expression of TLR4 protein. In these cells, IFN-gamma does not induce LPS responsiveness. The second phenotype, which includes cell line SW480, exhibits a highly LPS-responsive phenotype and surface expression of TLR4 protein even in unprimed conditions. These lines are functionally similar to cells of monocytic lineage. In the third phenotype, which includes the HT-29 and Colo205 cell lines, TLR4 protein is largely present in the cytoplasmic fraction and the cells are hyporesponsive to LPS in an unprimed condition. However, priming of these cells with IFN-gamma can induce LPS responsiveness through augmentation of LPS uptake and expression of MD-2 mRNA and intracellular TLR4 proteins. Finally, these findings suggest that the Th1 cytokine IFN-gamma modulates LPS responsiveness through several mechanisms in intestinal epithelial cells and that these cells may comprise different subpopulations with distinct roles in innate immune responses. PMID- 12761137 TI - CD40 contributes to lethality in acute sepsis: in vivo role for CD40 in innate immunity. AB - Sepsis induces an early inflammatory cascade initiated by the innate immune response. This often results in the development of multisystem organ failure. We examined the role of CD40, a costimulatory molecule that is integral in adaptive immunity, by using a murine model of polymicrobial sepsis. CD40 knockout (KO) mice had delayed death and improved survival after cecal ligation and puncture (CLP). In addition, they had less remote organ injury as manifested by reduced pulmonary capillary leakage. The improvements in survival and remote organ dysfunction in CD40 KO mice were associated with reduced interleukin-6 (IL-6) and IL-10 levels in serum and bronchoalveolar lavage fluid compared to the levels in wild-type (WT) controls. Furthermore, in contrast to WT mice, CD40 KO mice had no induction of the Th1 cytokines IL-12 and gamma interferon in serum or lungs after CLP. The alterations in cytokine production in CD40 KO mice were associated with similar changes in transcription factor activity. After CLP, CD40 KO mice had attenuated activation of nuclear factor kappaB and signal transducer and activator of transcription 3 in both the lung and the liver. Finally, WT mice had increased expression of CD40 on their alveolar macrophages. These data highlight the importance of CD40 activation in the innate immune response during polymicrobial sepsis and the subsequent development of remote organ dysfunction. PMID- 12761139 TI - Drosophila melanogaster is a genetically tractable model host for Mycobacterium marinum. AB - Mycobacterium marinum is a pathogenic mycobacterial species that is closely related to Mycobacterium tuberculosis and causes tuberculosis-like disease in fish and frogs. We infected the fruit fly Drosophila melanogaster with M. marinum. This bacterium caused a lethal infection in the fly, with a 50% lethal dose (LD(50)) of 5 CFU. Death was accompanied by widespread tissue damage. M. marinum initially proliferated inside the phagocytes of the fly; later in infection, bacteria were found both inside and outside host cells. Intracellular M. marinum blocked vacuolar acidification and failed to colocalize with dead Escherichia coli, similar to infections of mouse macrophages. M. marinum lacking the mag24 gene were less virulent, as determined both by LD(50) and by death kinetics. Finally, in contrast to all other bacteria examined, mycobacteria failed to elicit the production of antimicrobial peptides in Drosophila.We believe that this system should be a useful genetically tractable model for mycobacterial infection. PMID- 12761138 TI - pH-regulated gene expression of the gastric pathogen Helicobacter pylori. AB - Colonization by the gastric pathogen Helicobacter pylori has been shown to be intricately linked to the development of gastritis, ulcers, and gastric malignancy. Little is known about mechanisms employed by the bacterium that help it adapt to the hostile environment of the human stomach. In an effort to extend our knowledge of these mechanisms, we utilized spotted-DNA microarrays to characterize the response of H. pylori to low pH. Expression of approximately 7% of the bacterial genome was reproducibly altered by shift to low pH. Analysis of the differentially expressed genes led to the discovery that acid exposure leads to profound changes in motility of H. pylori, as a larger percentage of acid exposed bacterial cells displayed motility and moved at significantly higher speeds. In contrast to previous publications, we found that expression of the bacterial virulence gene cagA was strongly repressed by acid exposure. Furthermore, this transcriptional repression was reflected at the level of protein accumulation in the H. pylori cell. PMID- 12761140 TI - Catalases of Aspergillus fumigatus. AB - Upon infection of a host, the pathogenic fungus Aspergillus fumigatus is attacked by the reactive oxygen species produced by phagocytic cells. Detoxification of hydrogen peroxide by catalases was proposed as a way to overcome this host response. A. fumigatus produces three active catalases; one is produced by conidia, and two are produced by mycelia. The mycelial catalase Cat1p was studied previously. Here we characterized the two other catalases, their genes, and the phenotypes of gene-disrupted mutants. CatAp, a spore-specific monofunctional catalase, is resistant to heat, metal ions, and detergent. This enzyme is a dimeric protein with 84.5-kDa subunits. The 749-amino-acid polypeptide exhibits high levels of similarity to the Aspergillus nidulans CatA catalase and to bacterial catalase HPII of Escherichia coli. In spite of increased sensitivity to H(2)O(2), killing of DeltacatA conidia by alveolar macrophages and virulence in animals were similar to the killing of conidia by alveolar macrophages and virulence in animals observed for the wild type. In contrast to the Cat1p and CatAp catalases, the mycelial Cat2p enzyme is a bifunctional catalase-peroxidase and is sensitive to heat, metal ions, and detergent. This enzyme, an 82-kDa monomer, is homologous to catalase-peroxidases of several fungi and bacteria. Surprisingly, mycelium of the double Deltacat1Deltacat2 mutant with no catalase activity exhibited only slightly increased sensitivity to H(2)O(2) and was as sensitive to killing by polymorphonuclear neutrophils as mycelium of the wild type strain. However, this mutant exhibited delayed infection in the rat model of aspergillosis compared to infection by the wild-type strain. These results indicate that conidial catalase is not a virulence factor and that mycelial catalases transiently protect the fungus from the host. PMID- 12761141 TI - Interleukin-12 p40 secretion by cutaneous CD11c+ and F4/80+ cells is a major feature of the innate immune response in mice that develop Th1-mediated protective immunity to Schistosoma mansoni. AB - Radiation-attenuated (RA) schistosome larvae are potent stimulators of innate immune responses at the skin site of exposure (pinna) that are likely to be important factors in the development of Th1-mediated protective immunity. In addition to causing an influx of neutrophils, macrophages, and dendritic cells (DCs) into the dermis, RA larvae induced a cascade of chemokine and cytokine secretion following in vitro culture of pinna biopsy samples. While macrophage inflammatory protein 1alpha and interleukin-1beta (IL-1beta) were produced transiently within the first few days, the Th1-promoting cytokines IL-12 and IL 18 were secreted at high levels until at least day 14. Assay of C3H/HeJ mice confirmed that IL-12 secretion was not due to lipopolysaccharide contaminants binding Toll-like receptor 4. Significantly, IL-12 p40 secretion was sustained in pinnae from vaccinated mice but not in those from nonprotected infected mice. In contrast, IL-10 was produced from both vaccinated and infected mice. This cytokine regulates IL-12-associated dermal inflammation, since in vaccinated IL 10(-/-) mice, pinna thickness was greatly increased concurrent with elevated levels of IL-12 p40. A significant number of IL-12 p40(+) cells were detected as emigrants from in vitro-cultured pinnae, and most were within a population of rare large granular cells that were Ia(+), consistent with their being antigen presenting cells. Labeling of IL-12(+) cells for CD11c, CD205, CD8alpha, CD11b, and F4/80 indicated that the majority were myeloid DCs, although a proportion were CD11c(-) F4/80(+), suggesting that macrophages were an additional source of IL-12 in the skin. PMID- 12761142 TI - Helicobacter-induced chronic active lymphoid aggregates have characteristics of tertiary lymphoid tissue. AB - Susceptible strains of mice that are naturally or experimentally infected with murine intestinal helicobacter species develop hepatic inflammatory lesions that have previously been described as chronic active hepatitis. The inflammatory infiltrates in some models of chronic autoimmunity or inflammation resemble tertiary lymphoid organs hypothesized to arise by a process termed lymphoid organ neogenesis. To determine whether hepatic inflammation caused by infection with helicobacter could give rise to tertiary lymphoid organs, we used fluorescence activated cell sorting, immunohistochemistry, and in situ hybridization techniques to identify specific components characteristic of lymphoid organs in liver tissue sections and liver cell suspensions from helicobacter-infected mice. Small venules (high endothelial venules [HEVs]) in inflammatory lesions in Helicobacter species-infected livers were positive for peripheral node addressin. Mucosal addressin cell adhesion molecule also stained HEVs and cells with a staining pattern consistent with scattered stromal cells. The chemokines SLC (CCL 21) and BLC (CXCL13) were present, as were B220-positive B cells and T cells. The latter included a naive (CD45lo-CD62Lhi) population. These findings suggest that helicobacter-induced chronic active hepatitis arises through the process of lymphoid organ neogenesis. PMID- 12761143 TI - Intracellular replication of Mycobacterium marinum within Dictyostelium discoideum: efficient replication in the absence of host coronin. AB - Mycobacterium marinum causes tuberculosis-like disease in fish and amphibians and has been used as a model mycobacterial species because of its rapid growth and less stringent containment requirements relative to other mycobacterial species. We demonstrate here that M. marinum grows within Dictyostelium discoideum cells, allowing the genetic analysis of host factors that may modulate the replication of mycobacterial species. Intracellular growth of M. marinum was shown to mimic the properties previously observed for growth within cultured phagocytes. A defined bacterial mutant defective for growth within phagocytic cells was shown to be similarly defective for growth within D. discoideum. To test the role of host coronin, which was previously hypothesized to positively modulate mycobacterial growth within mouse macrophages, a defined D. discoideum coronin mutant was analyzed. Surprisingly, the absence of coronin resulted in enhanced intracellular replication of M. marinum relative to the control wild-type strain. Consistent with previous observations, some phagosomes showed persistence of coronin about the surface of the compartment, but colocalization of the protein was far from uniform. We conclude that in D. discoideum factors other than coronin support intracellular replication of M. marinum. PMID- 12761144 TI - Analysis of the OspE determinants involved in binding of factor H and OspE targeting antibodies elicited during Borrelia burgdorferi infection in mice. AB - Immune evasion by Lyme spirochetes is a multifactorial process involving numerous mechanisms. The OspE protein family undergoes antigenic variation during infection and binds factor H (fH) and possibly FHL-1/reconectin. In Borrelia burgdorferi B31MI, the OspE family consists of three paralogs: BBL39 (ErpA), BBP38, and BBN38 (ErpP). BBL39 and BBP38 are identical and therefore are referred to here as BBL39. The goals of this study were to assess the specificity of the antibody (Ab) response to the OspE paralogs and to identify the domains or determinants of OspE that are required for the binding of fH and OspE-targeting Abs that develop during infection. Here we demonstrate that at least some of the anti-OspE Abs produced during infection are paralog specific and that Ab binding requires conformational determinants whose formation requires both the N- and C terminal domains of OspE. The binding of fH to OspE was also found to be dependent on conformational determinants. It is also demonstrated here that all of the OspE paralogs expressed by B. burgdorferi B31MI are capable of binding fH. The binding of fH to members of the OspF protein family was also assessed. In contrast to an earlier report, no binding of BBO39 or BBR42 to human fH was detected. Lastly, a series of competitive binding enzyme-linked immunosorbent assay analyses, designed to determine if fH and infection serum Abs bind to the same sites on OspE, revealed that these ligands interact with different regions of OspE. PMID- 12761145 TI - Comprehensive analysis of the factor h binding capabilities of borrelia species associated with lyme disease: delineation of two distinct classes of factor h binding proteins. AB - Some Lyme disease spirochete isolates can bind complement regulatory protein factor H (fH), a process that may allow evasion of complement-mediated killing. Here we demonstrate significant differences in the fH binding capabilities of species of the Borrelia burgdorferi sensu lato complex. The percentages of B. burgdorferi, B. afzelii, and B. garinii bacteria that bound fH in either enzyme linked immunosorbent assays or affinity ligand binding immunoblot assays were 100, 83, and 29%, respectively. The fH binding protein profiles were examined and found to exhibit variability among isolates and to form two distinct classes. Differences in fH binding ability may contribute to the differences in pathogenesis and clinical course observed upon infection with different species of the B. burgdorferi sensu lato complex. PMID- 12761146 TI - A dominant role for extracellular glutathione S-transferase from Onchocerca volvulus is the production of prostaglandin D2. AB - The extracellular glutathione S-transferase from the filarial parasite Onchocerca volvulus (Ov-GST1) is a glutathione-dependent prostaglandin D synthase. Ov-GST1, located in the outer hypodermal lamellae and in parts of the cuticle, produces prostaglandin D(2) directly at the parasite-host interface. Ov-GST1 therefore has the potential to participate in the modulation of the host immune response by contributing to the production of prostanoids; this supports the predominant hypothesis that parasite-derived eicosanoids influence host inflammatory and immune cells. PMID- 12761147 TI - Visualization of Proteus mirabilis morphotypes in the urinary tract: the elongated swarmer cell is rarely observed in ascending urinary tract infection. AB - Proteus mirabilis, a common cause of nosocomial and catheter-associated urinary tract infection, colonizes the bladder and ascends the ureters to the proximal tubules of the kidneys, leading to the development of acute pyelonephritis. P. mirabilis is capable of swarming, a form of multicellular behavior in which bacteria differentiate from the short rod typical of members of the family Enterobacteriaceae, termed the swimmer cell, into hyperflagellated elongated bacteria capable of rapid and coordinated population migration across surfaces, called the swarmer cell. There has been considerable debate as to which morphotype predominates during urinary tract infection. P. mirabilis(pBAC001), which expresses green fluorescent protein in both swimming and swarming morphotypes, was constructed to quantify the prevalence of each morphotype in ascending urinary tract infection. Transurethral inoculation of P. mirabilis(pBAC001) resulted in ascending urinary tract infection and kidney pathology in mice examined at both 2 and 4 days postinoculation. Using confocal microscopy, we were able to investigate the morphotypes of the bacteria in the urinary tract. Of 5,087 bacteria measured in bladders, ureters, and kidneys, only 7 (0.14%) were identified as swarmers. MR/P fimbria expression, which correlates with the swimmer phenotype, is prevalent on bacteria in the ureters and bladder. We conclude that, by far, the predominant morphotype present in the urinary tract during ascending infection is the short rod-the swimmer cell. PMID- 12761148 TI - Listeriolysin O-mediated calcium influx potentiates entry of Listeria monocytogenes into the human Hep-2 epithelial cell line. AB - To investigate factors which modulate the entry of Listeria monocytogenes into mammalian cells, we have analyzed the role of Ca(2+). We show that L. monocytogenes induced Ca(2+) transients into the human Hep-2 epithelial cell line. The nonpathogenic species L. innocua or a L. monocytogenes mutant strain defective in listeriolysin O (LLO) production was unable to induce these calcium fluxes. Addition of plasma membrane calcium channel antagonists or chelation of extracellular calcium markedly reduced L. monocytogenes entry. In contrast, chelation of host cytosolic Ca(2+) or blockade of Ca(2+) release from intracellular stores did not affect invasion. These results indicate that L. monocytogenes-induced mobilization of extracellular Ca(2+) by LLO and activation of downstream Ca(2+)-dependent signaling are required for efficient cell invasion. PMID- 12761149 TI - The FimH A27V mutation is pathoadaptive for urovirulence in Escherichia coli B2 phylogenetic group isolates. AB - Correlations between FimH mutations and virulence were established by studying a collection of human commensal and extraintestinal pathogenic Escherichia coli natural isolates. Pathoadaptive (A27V and, to a lesser extent, A119V) and "commensal-adaptive" (A202V) mutations were evidenced in B2 phylogenetic group strains. fimH phylogenetic analysis indicates that these pathoadaptive mutations occurred several times. PMID- 12761150 TI - Helicobacter pylori supernatants cause epithelial cytoskeletal disruption that is bacterial strain and epithelial cell line dependent but not toxin VacA dependent. AB - We show here that Helicobacter pylori broth culture supernatants disrupt the actin cytoskeleton of epithelial cell lines, leading to cell rounding and apoptosis through anoikis. We demonstrate that there are marked quantitative differences between strains and that there are different cell line sensitivities. By constructing VacA null isogenic mutants, we show that the effect is not due to the vacuolating cytotoxin. PMID- 12761152 TI - Cytolethal distending toxin gene cluster in enterohemorrhagic Escherichia coli O157:H- and O157:H7: characterization and evolutionary considerations. AB - We identified a cytolethal distending toxin (cdt) gene cluster in 87, 6, and 0% of sorbitol-fermenting (SF) enterohemorrhagic Escherichia coli (EHEC) O157:H(-), EHEC O157:H7, and E. coli O55:H7/H(-) strains, respectively. The toxin was expressed by the wild-type EHEC O157 strains and by a cdt-containing cosmid from a library of SF EHEC O157:H(-) strain 493/89. The cdt flanks in strain 493/89 were homologous to bacteriophages P2 and lambda. Our data demonstrate that cdt, encoding a potential virulence factor, is present in the EHEC O157 complex and suggest that cdt may have been acquired by phage transduction. PMID- 12761153 TI - Complete sequence of the cap locus of Haemophilus influenzae serotype b and nonencapsulated b capsule-negative variants. AB - The complete capsule (cap) loci from three Haemophilus influenzae strains, one serotype b (Hib) and two nonencapsulated b capsule-negative variants, were sequenced. Two new open reading frames, hcsA and hcsB, were identified in region III and thought to be involved in postpolymerization modification of the capsule. The location of the cap locus in the Haemophilus influenzae chromosome was identified within section 97 of the Rd genome (chromosomal coordinates 1074542 to 1086327) and found to be the same for the Hib and two Hib(-) strains as well as some other encapsulated division I H. influenzae strains. PMID- 12761151 TI - Vaccine-induced protection against Helicobacter pylori in mice lacking both antibodies and interleukin-4. AB - To test the hypothesis that a Th2 response to Helicobacter pylori is necessary for protection and to address the possibility that humoral and Th2 cellular responses may compensate for each other, we generated mice deficient in both interleukin-4 (IL-4) and antibodies. The immunized double-knockout mice were protected from H. pylori challenge, as were the parental strains and wild-type C57BL/6 mice. Neutralization of IL-4 in B-cell-deficient mice did not prevent protection. Immunized IL-5-deficient mice were also protected. Thus, IL-4 and IL 5 are not essential for protection. PMID- 12761154 TI - Involvement of nicotinic acetylcholine receptors in controlling Chlamydia pneumoniae growth in epithelial HEp-2 cells. AB - Nicotinic acetylcholine receptors (nAChRs) play an essential role in neurotransmission. Recent studies have indicated that nAChRs may be involved in the regulation of some bacterial infections through immunological mechanisms in macrophages. However, the regulation of infection with Chlamydia pneumoniae, which is a ubiquitous pneumonia-causing bacterium, by an nAChR-mediated mechanism is still unclear. In the present study, it was found that stimulation of nAChRs with ligands such as nicotine and acetylcholine altered the growth of C. pneumoniae in epithelial HEp-2 cells. Thus, the results revealed a possible pathophysiological role of nAChRs in the regulation of intracellular bacterial infection. PMID- 12761155 TI - Chemokine receptor CCR2 is not essential for the development of experimental cerebral malaria. AB - Infection with Plasmodium berghei ANKA induces cerebral malaria in susceptible mice. Brain-sequestered CD8(+) T cells are responsible for this pathology. We have evaluated the role of CCR2, a chemokine receptor expressed on CD8(+) T cells. Infected CCR2-deficient mice were as susceptible to cerebral malaria as wild-type mice were, and CD8(+) T-cell migration to the brain was not abolished. PMID- 12761156 TI - Actinobacillus actinomycetemcomitans serotype b lipopolysaccharide mediates coaggregation with Fusobacterium nucleatum. AB - Purified Actinobacillus actinomycetemcomitans serotype b lipopolysaccharide (LPS) was found to be able to bind Fusobacterium nucleatum cells and to inhibit binding of F. nucleatum to A. actinomycetemcomitans serotype b. Sugar binding studies showed that the requirements for binding of A. actinomycetemcomitans serotype b LPS to the F. nucleatum lectin are the presence of a metal divalent ion, an axial free hydroxyl group at position 4, and free equatorial hydroxyl groups at positions 3 and 6 of D-galactose, indicating that the beta-N-acetyl-D galactosamine in the serotype b LPS trisaccharide repeating unit is the monosaccharide residue recognized by the F. nucleatum lectin. These data strongly suggest that A. actinomycetemcomitans serotype b LPS is one of the receptors responsible for the lactose-inhibitable coaggregation of A. actinomycetemcomitans to fusobacteria. PMID- 12761157 TI - Physical map and genome sequencing survey of Mycoplasma haemofelis (Haemobartonella felis). AB - Mycoplasma haemofelis is an uncultivable red-cell pathogen of cats. Isolated M. haemofelis DNA was used to create a bacterial artificial chromosome library and physical map. Random sequencing of this material revealed 75 genes that had not been previously reported for M. haemofelis or any other hemotrophic mycoplasma. PMID- 12761158 TI - The stereochemistry of the amino acid side chain influences the inflammatory potential of muramyl dipeptide in experimental meningitis. AB - Intrathecal injections of 50 to 100 micro g of (N-acetylmuramyl-L-alanyl-D isoglutamine) muramyl dipeptide (MDP)/rabbit dose-dependently triggered tumor necrosis factor alpha (TNF-alpha) secretion (12 to 40,000 pg/ml) preceding the influx of leukocytes in the subarachnoid space of rabbits. Intrathecal instillation of heat-killed unencapsulated R6 pneumococci produced a comparable leukocyte influx but only a minimal level of preceding TNF-alpha secretion. The stereochemistry of the first amino acid (L-alanine) of the MDP played a crucial role with regard to its inflammatory potential. Isomers harboring D-alanine in first position did not induce TNF-alpha secretion and influx of leukocytes. This stereospecificity of MDPs was also confirmed by measuring TNF-alpha release from human peripheral mononuclear blood cells stimulated in vitro. These data show that the inflammatory potential of MDPs depends on the stereochemistry of the first amino acid of the peptide side chain and suggest that intact pneumococci and MDPs induce inflammation by different pathways. PMID- 12761159 TI - Natural history of Helicobacter hepaticus infection in conventional A/J mice, with special reference to liver involvement. AB - It has been reported that Helicobacter hepaticus infection of mice leads to chronic hepatitis and hepatocarcinoma. Our aim was to monitor a cohort of 80 conventional A/J mice in which half of the mice were infected by H. hepaticus in order to study the evolution of the infection and the pathological changes in comparison to uninfected mice. H. hepaticus was detected by culture only in some colon and cecum specimens after 17 months of age, while PCR detected H. hepaticus in the intestines of all inoculated mice after only 5 months of infection. The percentage of mice in which H. hepaticus was detected in the gallbladder, bile ducts, and liver by PCR, as well as the number of bacteria present in the liver, tended to increase with increasing age and longer infection time. Anti-H. hepaticus immunoglobulin G antibodies were positive by enzyme-linked immunosorbent assay only in inoculated mice. Pathological findings were also more frequent as the mice grew older: fibrosis was present (especially in the peripheral part of the liver), and significant portal inflammation including lymphoid nodules was present in almost all infected animals. Biliary lesions of neutrophilic acute cholangitis or lymphocytic cholangitis were noted. However, lesions were also observed in uninfected animals, although at a significantly lower level, and the only hepatocellular carcinoma occurred in an uninfected mouse. The evolution towards hepatocarcinoma is not always the endpoint and may depend on the bacterial strain and on the environmental conditions. PMID- 12761160 TI - RNA aptamers targeted to domain II of hepatitis C virus IRES that bind to its apical loop region. AB - The internal ribosome entry site (IRES) is important for translation of hepatitis C virus (HCV) mRNA and has a unique RNA structure containing conserved domains I to IV. To investigate the function of domain II, we selected RNA aptamers that bind to domain II of HCV IRES by applying a simple and convenient selection method using a hybridized tag for fixing domain II RNA on magnetic beads instead of synthesizing long RNA. In addition, we employed surface plasmon resonance (SPR) technology to measure the binding affinity of each generation and to obtain detailed kinetic constants. The selected aptamers have a consensus sequence, 5' UAUGGCU-3', which is complementary to the apical loop of domain II. The loop-loop interaction between the consensus sequence and domain II was confirmed by mutagenesis and nuclease mapping analyses. Binding affinities were dependent on the local structure containing the conserved sequence. The aptamers could inhibit IRES-dependent translation. PMID- 12761161 TI - SRCL/CL-P1 recognizes GalNAc and a carcinoma-associated antigen, Tn antigen. AB - SRCL /CL-P1 was recently identified as a scavenger receptor with a C-type lectin domain, which was expressed in vascular endothelial cells and could bind to Gram positive and Gram-negative bacteria, yeast and oxidized LDL. We found that SRCL was expressed in some but not all nurse-like cells examined. Furthermore, to characterize the C-type lectin domain of SRCL, the secreted form of the C-type lectin domain (LEC-AP) of SRCL, which was fused to the signal sequence of IgG and alkaline phosphatase, was expressed in 293/EBNA-1 cells and the culture medium was used for the in vitro binding assay. LEC-AP specifically bound to GalNAc conjugated gel in a Ca(2+)-dependent manner, and this binding was inhibited by free GalNAc, L-, D-fucose, D-galactose, lactose, and especially T antigen and Tn antigen. Furthermore, we examined whether or not SRCL could take up saccharide conjugated particles. 293/EBNA-1 cells stably expressing SRCL were found to take up GalNAc but not mannose-conjugated particles on confocal microscopy. The binding of GalNAc-conjugated particles to these cells was quantitatively measured by comparing the x-means of individual cell populations. An approximately 2.1 fold increase in immunofluorescence intensity was observed for the SRCL transfectants compared to control vector transfectants. Our results provide a basis for understanding the scavenger function of SRCL as to carbohydrate containing ligands. PMID- 12761162 TI - Glu-53 of Bacillus cereus sphingomyelinase acts as an indispensable ligand of Mg2+ essential for catalytic activity. AB - Bacillus cereus sphingomyelinase (SMase) is an extracellular hemolysin classified into a group of Mg(2+)-dependent neutral SMases (nSMase). Sequence comparison of bacterial and eukaryotic Mg(2+)-dependent nSMases has shown that several amino acid residues, including Glu-53 of B. cereus SMase, are conserved, suggesting a catalytic mechanism common to these enzymes. Mutational analysis has revealed that hemolytic and SM-hydrolyzing activities are abolished by E53A and E53Q mutations. Only the E53D mutant enzyme partially retains these activities, however, a significant decrease in the apparent k(cat)/K(m) for SM hydrolysis is observed by this mutation. Mg(2+) activates the wild-type enzyme in a two-step manner, i.e., at least two binding sites for Mg(2+), high- and low-affinity, are present on the enzyme. The binding affinity of essential Mg(2+) for the high affinity site is decreased by the mutation. In addition, the binding affinities of Mn(2+) and Co(2+) (substitutes for Mg(2+)) are also decreased. On the contrary, the inhibitory effects of Ca(2+), Cu(2+), and Zn(2+) on SM-hydrolyzing activity are not influenced by the mutation. The results indicate that Glu-53 of B. cereus SMase acts as a ligand for Mg(2+) and is involved in the high-affinity Mg(2+)-binding site, which is independent of the binding site for inhibitory metals. PMID- 12761163 TI - N-acetylglucosamine-6-O-sulfotransferase-1: production in the baculovirus system and its applications to the synthesis of a sulfated oligosaccharide and to the modification of oligosaccharides in fibrinogen. AB - N-acetylglucosamine-6-O-sulfotransferase (GlcNAc6ST) catalyzes the transfer of sulfate from 3'-phosphoadenosine 5'-phosphosulfate to the C-6 position of non reducing GlcNAc. Human GlcNAc6ST-1 was expressed as a fusion protein with protein A in an insect cell line (Tn 5 cells) using the baculovirus system. The recombinant enzyme was purified to homogeneity by IgG Sepharose column chromatography. The substrate specificity and the kinetic properties of the enzyme were similar to those of the enzyme expressed in the mammalian system. The purified recombinant enzyme was used to synthesize 6-sulfo GlcNAcbeta1-3Galbeta1 4Glc, which was identified by time of flight mass spectrometry. This sulfated trisaccharide served as a better substrate for microsomal galactosyltransferase from the mouse colon compared to 6-sulfo GlcNAc. The purified recombinant enzyme was also used to sulfate oligosaccharide chains on fibrinogen after enzymatic desialylation and degalactosylation to expose nonreducing GlcNAc residues. It is known that desialylation greatly increases the rate of clotting of fibrinogen after the addition of thrombin. Subsequent sulfation of desialylated and degalactosylated fibrinogen slightly decreased the rate of clotting. The recombinant GlcNAc6ST-1 is a useful reagent for 6-sulfate exposed GlcNAc residues both in oligosaccharides and in glycoproteins. PMID- 12761164 TI - Involvement of ClpX protein in the post-transcriptional regulation of a competence specific transcription factor, ComK protein, of Bacillus subtilis. AB - ComK protein of Bacillus subtilis positively regulates the transcription of several late competence genes as well as comK itself. We constructed a clpX disrupted mutant of B. subtilis and studied its effect on the regulation of ComK activation. When Pspac, which controls the comK gene in a multicopy plasmid, was induced by the addition of IPTG, comK transcripts were detected in both the clpX mutant and the wild type. However, the ComK protein could not be detected in the clpX disrupted mutant. To obtain further information, we constructed several comK lacZ translational fusions covering different lengths of the comK gene, whose transcription is controlled by an IPTG inducible Pspac promoter. We found that both the expression of comK-lacZ directed beta-galactosidase and the accumulation of ComK-LacZ fused protein, derived from the fusion containing the entire comK open reading frame, were extremely reduced in the clpX mutant compared with the wild type, while the accumulation of comK-lacZ transcripts in the clpX mutant after the addition of IPTG was about half that in the clpX+ background. On the other hand, transcription, translation and activity of comK-lacZ were detected in both the clpX mutant and the wild type when the comK-lacZ fusion lacking the 3' region of the comK gene was induced. These results indicate that ClpX plays an important role in the regulation of ComK at the post-transcriptional level. PMID- 12761165 TI - Significant reduction of WT1 gene expression, possibly due to epigenetic alteration in Wilms' tumor. AB - WT1 at 11p13 is a tumor suppressor gene, an aberration of which causes Wilms' tumor (WT). Since WT1 expression is reduced in a certain proportion of WTs and its mutation is found only in 10-20% of WTs, we examined WT1 gene silencing due to epigenetic alteration in a total of 22 WTs. WT1 expression was significantly reduced in half of WTs without any mutation in the WT1 gene itself, suggesting that the reduction of expression was possibly epigenetic. We found promoter hypermethylation in one WT with loss of heterozygosity (LOH) and showed that promoter methylation reduced reporter gene activity by a reporter assay. These data suggested that methylation was an epigenetic mechanism leading to WT1 silencing and that the expression-reduced allele by hypermethylation combined with LOH was consistent with the revised two-hit model. In addition, as the beta catenin mutation is frequently associated with the WT1 mutation, the association of WT1 silencing with the beta-catenin mutation was also investigated. beta catenin mutated in only one WT without WT1 silencing, suggesting that the beta catenin mutation was not associated with the reduction of WT1 expression. PMID- 12761166 TI - 5S rRNA is a leadzyme. A molecular basis for lead toxicity. AB - This paper reports that the D-loop sequence of cellular mammalian ribosomal 5S RNAs is a natural leadzyme that specifically binds and cleaves in trans other RNA molecules in the presence of lead. The D-loops of these 5S rRNAs are similar in sequence to the active site of the leadzyme derived from tRNA(Phe), which cleaves a single bond in cis. We have devised a 12 nt model substrate based on the leadzyme sequence cleaved in trans by a 12 nt RNA molecule containing of the D loop sequence. The model reaction occurs only at the appropriate concentration of lead and enzyme/substrate stoichiometry. The native 5S rRNA carries the same cleavage activity, although with different optimal lead concentration and stoichiometry. On the other hand, the isolated D-loop does not serve as a substrate when incubated with an RNA molecule with the potential to base pair with it and form the same internal loop (the bubble) present in the leadzyme substrate complex. We show that the leadzyme cuts C-G, but not G-G or U-G linkages. The 5S rRNA leadzyme appears to have the shortest asymmetric pentanucleotide purine-rich loop flanked by two short double stranded RNAs. The leadzyme activity of native 5S rRNA may be an important aspect of lead toxicity in living cells. Because the leadzyme motif has been found in natural RNA species, its activity can be expressed in vivo even at a very low lead concentrations, of lead leading to the inactivation of other cellular RNAs. This might be one of the ways in which lead poisoning manifests itself at the molecular level. Lead toxicity is based not only on its binding to calcium and zinc binding proteins (such as Zn-fingers) and random hydrolysis of nucleic acids, but also, and most importantly, on the induction of the hydrolytic properties of RNA (RNA catalysis). PMID- 12761167 TI - Biochemical and genetic analyses of a novel gamma-cyclodextrin glucanotransferase from an alkalophilic Bacillus clarkii 7364. AB - On screening for microorganisms in soil obtained in Japan that produce large amounts of gamma-cyclodextrin (gamma-CD), we identified a novel alkalophilic bacterium, Bacillus clarkii 7364. The cyclodextrin glucanotransferase (CGTase) secreted into the culture medium by this bacterium was purified by affinity chromatography on a gamma-CD-immobilized column, followed by chromatography on a gel filtration column. The enzyme converted 13.7% of pre-gelatinized potato starch (10% w/w per reaction mixture) into CDs, and the majority (79%) of the product CDs was of the gamma form. This property is quite unique among known CGTases and thus we named this enzyme gamma-CGTase. The N-terminal and internal amino acid sequences of gamma-CGTase were determined and used to design PCR primers for amplification of the nucleotide sequence that encodes the gamma CGTase gene. The entire gene sequence amplified by PCR was determined and then cloned into E. coli. The recombinant enzyme synthesized by E. coli retained biochemical properties quite similar to those of the original one. Comparison of the deduced amino acid sequence of gamma-CGTase with those of other known CGTases that have different product specificities revealed the importance of subsites -3 and -7 for the preferential gamma-cyclization activity. PMID- 12761168 TI - Identification of the carboxyl-terminal membrane-anchoring region of HPC 1/syntaxin 1A with the substituted-cysteine-accessibility method and monoclonal antibodies. AB - HPC-1/syntaxin 1A is a member of the syntaxin family, and functions at the plasma membrane during membrane fusion as the target-soluble N-ethylmaleimide-sensitive factor-attachment protein receptor (t-SNARE). We identified the membrane anchoring region of HPC-1/syntaxin 1A, and examined its role in anchoring of a protein to the plasma membrane. A series of mutants was created from a cysteine less mutant of HPC-1/syntaxin 1A by substitution of each residue at the C terminus with cysteine. The accessibility of the thiol-groups in each mutant was analyzed in vivo. The cysteine (C145) within the N-terminal cytosolic segment was labeled, but not that at C271 or C272, or any of those introduced at the C terminus. The addition of additional residues to the C-terminal tail of HPC 1/syntaxin 1A allowed labeling by thiol-specific reagents. A monoclonal antibody directed against the C-terminal tail peptide did not react with the protein located at the plasma membrane. In addition, subcellular fractionation and immunocytochemical analyses with various transmembrane mutants showed that the C terminal tail comprising eight amino acids is essential for anchoring of HPC 1/syntaxin 1A to the plasma membrane. These results indicate that the C-terminal membrane-anchoring region, which comprises 23 amino acids, does not traverse the lipid-bilayer and that the C-terminal tail is essential for anchoring of HPC 1/syntaxin 1A to the plasma membrane. PMID- 12761170 TI - Anomalous excitonic CD of meso-tetrakis(3-N-methylpyridiniumyl)porphyrin bound to poly[d(A-T)(2)]. AB - When meso-tetrakis(3-N-methylpyridiniumyl)porphyrin (m-TMPyP) formed a complex with poly[d(A-T)(2)], an intense bisignate excitonic CD in the Soret absorption region was observed. The excitonic CD of the m-TMPyP-poly[d(A-T)(2)] complex is unique in that no other combination of the related porphyrin, namely, meso tetrakis(n-N-methylpyridiniumyl)porphyrin (where n = 2, 4), and polynucleotide including calf thymus DNA, poly[d(G-C)(2)], poly[d(I-C)(2)], and poly(dA).poly(dT), exhibits a comparable CD spectrum. From the [drug]/[DNA] ratio dependence of the intensity and the shape of the CD spectrum, this porphyrin species is assigned to an extensively aggregated form. The extensively aggregated porphyrin disperses in 1 h after mixing to form moderately stacked porphyrin at a low mixing ratio. The magnitude of linear dichroism of the extensively aggregated porphyrin was small and the sign was negative in the Soret band, which indicated that the molecular plane of porphyrin in the complex is strongly tilted. On the other hand, the molecular plane of porphyrin is almost parallel to the DNA base plane (perpendicular to the DNA helix axis) in the moderately stacked form. PMID- 12761169 TI - Theoretical research on structures of gamma-aminobutyric acid and glutamic acid in aqueous conditions. AB - Even though glutamic acid contains only one more carboxyl group than gamma aminobutyric acid (GABA), these neurotransmitters are recognized by their own specific receptors. To understand the ligand-recognition mechanism of the receptors, we must determine the geometric and electronic structures of GABA and glutamic acid in aqueous conditions using the ab initio calculation. The results of the present study showed that the stable structure of GABA was the extended form, and it attracted both cations and anions. Glutamic acid only attracted cations and was stabilized in four forms in aqueous conditions: Type 1 (an extended form), Type 2 (a rounded form), and Types 3 and 4 (twisted forms of Type 1). The former two types had low energy and the energy barrier between them was estimated to be small. These results showed that most free glutamic acid is present as Type 1, Type 2, and transient forms. The present results therefore suggest that the flexibility of the geometric structures of ligands should be taken into account when we attempt to elucidate the mechanism of recognition between ligands and receptors, in addition to the physicochemical characteristics of ligands and receptors. PMID- 12761171 TI - Structure/function analysis of an RNA aptamer for hepatitis C virus NS3 protease. AB - RNA aptamers that bind specifically to hepatitis C virus (HCV) NS3 protease domain (DeltaNS3) were identified in previous studies. These aptamers, G9-I, -II, and -III, were isolated using an in vitro selection method and they share a common loop with the sequence 5'-GA(A/U)UGGGAC-3'. The aptamers are potent inhibitors of the NS3 protease in vitro and may have potential as anti-HCV compounds. G9-I has a 3-way stem-loop structure and was selected for further characterization using site-directed mutagenesis. Mutations or deletions in stem loop II do not interfere with binding or inhibition of DeltaNS3, but mutations or deletions in stem I and stem-loop III destroy the G9-I active conformation and interfere with inhibition of NS3 protease. A 51 nt fragment of 74 nt G9-I was identified (DeltaNEO-III) as is the minimal fragment of G9-I that is an effective inhibitor of the NS3 protease. Tertiary interactions involving functionally important nucleotides were identified in the active structure of G9-I using nucleotide analog interference mapping (NAIM). Strong interferences were focused in the conserved loop involving stem-loop III and stem I. For example, analog interference caused at A(+8) and C(+24)-G(-36) base pair implied an A-minor motif involving the intramolecular base triple A(+8).C(+24)-G(-36), which is further supported by mutagenesis. These results suggested the interaction of stem I and stem-loop III is essential for the function of G9-I aptamer. PMID- 12761172 TI - Identification of 36-kDa flagellar phosphoproteins associated with hamster sperm motility. AB - In our previous paper [M. Fujinoki et al. (2001) BIOMED: Res. 22, 45-58], we reported that two types of 36-kDa protein, which were designated as 36K-A protein and 36K-B protein, obtained from hamster sperm flagella were phosphorylated at serine residues associated with the regulation of motility activation. In the present experiments, it was suggested that these two types of 36-kDa protein were phosphorylated in a cAMP-dependent manner associated with motility activation of hamster spermatozoa. Because the 36K-B protein was the most intensely phosphorylated in a cAMP-dependent manner, attempts were made to further characterize it. The 36K-B protein was assumed to be localized in the middle piece. The localization of the 36K-B protein was the same as that of the 36-kDa protein reported in our previous paper [Y. Si et al. (1999) Mol. Reprod. Dev. 52, 328-334]. In order to identify the 36K-B protein, it was analyzed by peptide mass finger printing and amino acid sequencing. The results suggested that the 36K-B protein was a pyruvate dehydrogenase E1 component beta subunit and a component of the mitochondrial sheath of the middle piece. PMID- 12761173 TI - Characterization of folding pathways of the type-1 and type-2 periplasmic binding proteins MglB and ArgT. AB - The family of periplasmic binding proteins (PBPs) is believed to have arisen from a common ancestor and to have differentiated into two types. At first approximation, both types of PBPs have the same fold pattern, reflecting their common origin. However, the connection between the main chains of a type 2 PBP is more complicated than a type 1 PBP's. We have been interested in the possibility that such structural changes affect the folding of PBPs. In this study, we have characterized the folding pathways of MglB (a type 1 PBP) and ArgT (a type 2 PBP) by using urea gradient gel electrophoresis, fast protein size-exclusion liquid chromatography and hydrophobic dye ANS binding assay. We found a distinct difference in folding between these two proteins. The folding of MglB followed a simple two-state transition model, whereas the folding of ArgT was more complicated. PMID- 12761174 TI - Carp hepatopancreatic DNase I: biochemical, molecular, and immunological properties. AB - A survey of DNase I in nine different carp tissues showed that the hepatopancreas has the highest levels of both DNase I enzyme activity and gene expression. Carp hepatopancreatic DNase I was purified 17,000-fold, with a yield of 29%, to electrophoretic homogeneity using three-step column chromatography. The purified enzyme activity was inhibited completely by 20 mM EDTA and a specific anti-carp DNase I antibody and slightly by G-actin. Histochemical analysis using this antibody revealed the strongest immunoreactivity in the cytoplasm of pancreatic tissue, but not in that of hepatic tissue in the carp hepatopancreas. A 995-bp cDNA encoding carp DNase I was constructed from total RNA from carp hepatopancreas. The mature carp DNase I protein comprises 260 amino acids, the same number as the human enzyme, however, the carp enzyme has an insertion of Ser59 and a deletion of Ala225 in comparison with the human enzyme. These alterations have no influence on the enzyme activity and stability. Three amino acid residues, Tyr65, Val67, and Ala114, of human DNase I are involved in actin binding, whereas those of carp DNase I are shifted to Tyr66, Val68, and Phe115, respectively, by the insertion of Ser59: the decrease in affinity to actin is due to one amino acid substitution, Ala114Phe. The results of our phylogenetic and immunological analyses indicate that carp DNase I is not closely related to the mammalian, avian or amphibian enzymes, and forms a relatively tight piscine cluster with the tilapia enzyme. PMID- 12761175 TI - Cloning and expression of the superoxide dismutase gene from the obligate anaerobic bacterium Desulfovibrio vulgaris (Miyazaki F). AB - We identified the SOD gene in the obligate anaerobic bacterium Desulfovibrio vulgaris (Miyazaki F) and constructed a high-level expression system in Escherichia coli. A 2.6-kbp DNA fragment isolated from D. vulgaris (Miyazaki F) by double digestion with EcoRI and SmaI contained the SOD gene and part of another open reading frame. The amino acid sequence deduced from the SOD gene, which was composed of 238 amino acid residues, showed high homogeneity with iron containing SOD (Fe-SOD) and predicted that the amino terminus of this protein would carry an export signal peptide. We produced the precursor form of SOD (PSOD) and the mature form of SOD (MSOD), which lacked the putative signal peptide. In E. coli, PSOD was present in insoluble inclusion bodies, and its putative signal peptide was not cleaved. In contrast, MSOD contained one iron per mononer and formed a dimer, which exhibited an SOD activity of 850 U/mg. Furthermore, D. vulgaris soluble extract showed a band of SOD activity on native polyacrylamide gel that migrated to the same point as MSOD. The intracellular localization of SOD and its role in D. vulgaris are also discussed. PMID- 12761176 TI - Discovery and prospect of protein kinase C research: epilogue. PMID- 12761177 TI - Identification and characterization of a novel calcineurin-binding protein in scallop testis. AB - Calcineurin has been inferred to function in meiosis and spermiogenesis in testis. Here, we identified a calcineurin-binding protein in scallop testis by Far-Western blot analysis using purified calcineurin as a probe. The molecular mass of the binding protein estimated on the blot was 75 kDa. The isolated cDNA clone encoded a novel 474-residue protein, named CaNBP75. The region between T6 and A210 of CaNBP75 was responsible for the interaction with calcineurin. CaNBP75 was predominantly expressed in testis and ovary of scallop. Thus, CaNBP75 may modulate the physiological function of calcineurin in the testis and ovary of scallop, such as in spermiogenesis or meiosis. PMID- 12761178 TI - Regulation of type 1 protein phosphatase/inhibitor-2 complex by glycogen synthase kinase-3beta in intact cells. AB - Inhibitor 2 (I-2) is a ubiquitous regulator of type 1 protein phosphatase (PP1). Previous in vitro studies suggested that its inhibitory activity towards PP1 is regulated by phosphorylation at Thr72 by glycogen synthase kinase-3beta (GSK 3beta), and at Ser86, Ser120, and Ser121 by casein kinase 2 (CK2). Here we report that GSK-3beta expressed in COS-7 cells phosphorylates wild-type I-2 but not an I 2 mutant carrying a T to A substitution at residue 72, showing that GSK-3beta phosphorylates I-2 at T72 in vivo as well. Co-immunoprecipitation study demonstrated that HA-GSK-3beta and I-2-FLAG co-exist in a same complex in the intact cells, but they do not bind directly. It is noteworthy that co-expression of Myc-PP1C significantly increased co-precipitation of HA-GSK-3beta with I-2 FLAG, showing a complex formation of HA-GSK-3beta/Myc-PP1C / I-2-FLAG in vivo. Further studies using a GSK-3beta kinase-dead mutant and LiCl, an inhibitor of GSK-3beta, showed that the enzyme activity of GSK-3beta is required for co precipitation. IP-Western study using several I-2 mutants substituted at phosphorylation sites (T72, S86, S120, and S121) suggested that phosphorylation of I-2 by CK2 is also involved in enhancement of association between GSK-3beta and I-2 in vivo. This study is the first demonstration that GSK-3beta associates with PP1C/I-2 complex and phosphorylates I-2 at T72 in the intact cells. PMID- 12761179 TI - A novel neutral amino acid transporter from the hyperthermophilic archaeon Thermococcus sp. KS-1. AB - A novel gene encoding a small neutral amino acid transporter was cloned from the genome of the hyperthermophilic archaeon Thermococcus sp. KS-1 by functional cloning using Escherichia coli strain AK430, which is defective in transporting glycine and D-alanine. The cloned gene, snatA, encoded a protein of 216 amino acid residues, SnatA, and was predicted to be a membrane protein with six membrane-spanning segments. E. coli AK430 cells transformed with snatA transported glycine with an apparent K(t) value of 24 micro M, which was one order of magnitude higher than that of other known glycine/alanine transporters, including cycA of E. coli and acp of thermophilic bacterium PS3. Competition studies revealed that SnatA transported various L-type neutral amino acids, but its substrate specificity was different from that of CycA or ACP. The glycine transport was inhibited by a protonophore, FCCP, or valinomycin plus nigericin, indicating that the process is dependent on an electrochemical potential of H(+). Homology searches revealed no homology with any transporters known to date. However, several hypothetical genes in prokaryote cells enrolled in the gene bank showed significantly high homology scores, indicating that snatA and its homologues form a family of prokaryotes. To our knowledge, this is the first report on the cloning of a gene of an amino acid transporter from a hyperthermophilic archaeon. PMID- 12761181 TI - Designed structural-rearrangement of an active group I ribozyme. AB - The catalytic core of the Tetrahyemena group I ribozyme consists of two functionally different domains, P4-P6 and P3-P7, that are conjugated via multiple tertiary interactions. The sequence encoding the P3-P7 domain is divided into two fragments in its primary sequence although the two domains are physically separable in the three dimensional (3D-) structure of the ribozyme: The sequence encoding the P4-P6 domain is inserted into that of the P3-P7 domain. An artificial rearrangement was designed and attempted for the primary sequence of the P3-P7 domain on the basis of a 3D-structural model and the biochemical data on the ribozyme. The domain in the primary structure was relocated to form a contiguous region while retaining the 3D-structure of the ribozyme required for self-splicing. The topologically rearranged ribozyme exhibited self-splicing activity. PMID- 12761180 TI - Regulation of a mitogen-activated protein kinase kinase kinase, MLTK by PKN. AB - PKNalpha is a fatty acid- and Rho-activated serine/threonine protein kinase having a catalytic domain homologous to members of the protein kinase C family. Recently it was reported that PKNalpha is involved in the p38 mitogen-activated protein kinase (MAPK) signaling pathway. To date, however, how PKNalpha regulates the p38gamma MAPK signaling pathway is unclear. Here we demonstrate that PKNalpha efficiently phosphorylates MLTKalpha (MLK-like mitogen-activated protein triple kinase), which was recently identified as a MAPK kinase kinase (MAPKKK) for the p38 MAPK cascade. Phosphorylation of MLTKalpha by PKNalpha enhances its kinase activity in vitro. Expression of the kinase-negative mutant of PKNalpha inhibited the mobility shift of MLTKalpha caused by osmotic shock in SDS-PAGE. Furthermore, PKNalpha associates with each member of the p38gamma MAPK signaling pathway (p38gamma, MKK6, and MLTKalpha). These results suggest that PKNalpha functions as not only an upstream activator of MLTKalpha but also a putative scaffold protein for the p38gamma MAPK signaling pathway. PMID- 12761182 TI - Extracellular cleavage of bullous pemphigoid antigen 180/type XVII collagen and its involvement in hemidesmosomal disassembly. AB - Bullous pemphigoid antigen 180 (BP180)/type XVII collagen is a transmembrane hemidesmosomal protein. Previously, we demonstrated that the collagenous ectodomain of BP180 can be cleaved within the extracellular non-collagenous (NC) 16A domain adjacent to the cell membrane and released from the cell surface. Here, we report that the BP180 cleavage is mediated by a membrane-associated metalloprotease expressed in epithelial cells. A tissue inhibitor of metalloprotease 1 (TIMP-1), but not TIMP-2, like the synthetic metalloprotease inhibitor KB-R8301, significantly reduced the cleavage. Within epithelial cells cultured for more than 36 h past confluency, antibodies to BP180 showed a reduced hemidesmosomal staining. Observed for the first time, addition of KB-R8301 to the cell culture preserved this staining. To examine the effect of the extracellular cleavage of BP180 on molecular interactions among hemidesmosomal components, we eliminated its collagenous extracellular portion, except for the NC16A domain, by collagenase digestion. Interestingly, this collagenase treatment caused partial disassembly of hemidesmosomal components in cultured human keratinocytes. Moreover, a monoclonal antibody specific for the cleaved extracellular fragment detected a unique tissue distribution of the fragment that might reflect an association of the cleavage process with the mitotic activity of epithelial tissues. Our observations demonstrate that the cleavage of BP180 occurring within the NC16A domain is mediated by a membrane-associated metalloprotease and suggest a possible involvement of the cleavage in hemidesmosomal disassembly. PMID- 12761183 TI - An x-ray diffraction study on the ADP-induced conformational change in skeletal muscle myosin. AB - Effects of ADP on the conformation of myosin cross-bridges were studied in x-ray diffraction experiments on single skinned fibers of frog skeletal muscle by photorelease of ADP from caged-ADP. The experiments were performed at the third generation synchrotron radiation facility SPring-8 with a time resolution of 5 ms. The intensity of the third-order meridional reflection from myosin filaments (at 1/14.4 nm(-1)) increased promptly after the ADP release with a time constant smaller than 5 ms, which was similar to that of tension decline. The results show that ADP binding induces a conformational change of myosin in skeletal muscle fibers. PMID- 12761184 TI - A presumed human nuclear autoantigen that translocates to plasma membrane blebs during apoptosis. AB - The structure and subcellular localization of a number of molecules change during apoptosis. These molecules are recognized by the immune system, leading to the development of autoimmunity when apoptotic cells fail to be effectively cleared by phagocytosis. We searched for such molecules by analyzing sera from 12 individuals who suffered from autoimmune diseases and from 3 patients with amyotrophic lateral sclerosis. One serum sample, designated 681, detected an antigen that fulfilled the above criteria. In Western blotting of lysates of human Jurkat T cells, the 681 antigen appeared as a distinct signal with a molecular mass of 60 kDa in normal cells, and 2 additional signals with faster mobilities were detected in apoptotic cells. The results of subcellular fractionation and immunofluorescence experiments revealed this antigen to be strictly localized in the nucleus of normal cells, but to be translocated to a region near the plasma membrane, to membrane blebs in particular, after the induction of apoptosis. Under conditions in which membrane blebbing was inhibited in apoptotic cells, the antigen still moved away from the nucleus, but its accumulation at the periplasmic region was completely abolished. The apparent partial cleavage and intracellular redistribution of the 681 antigen in apoptotic cells mimics changes previously reported for the nuclear autoantigen La, but the 681 antigen was clearly distinct from La. These results suggest that cleavage dependent exit from the nucleus during apoptosis is a phenomenon common to nuclear autoantigens. PMID- 12761185 TI - Cloning, expression, and characterization of the first archaeal ATP-dependent glucokinase from aerobic hyperthermophilic archaeon Aeropyrum pernix. AB - The gene encoding the ATP-dependent glucokinase of hyperthermophilic archaeon Aeropyrum pernix was identified, cloned, and functionally expressed in Escherichia coli. The deduced amino acid sequence showed 40% identity to that of the putative glucokinase from hyperthermophilic archaeon Pyrobacurum aerophilum. The purified recombinant enzyme was a monomer with a molecular mass of 35 kDa. The enzyme retained its full activity on heating at 70 degrees C for 10 min and retained 65% of the activity after 10-min incubation at 100 degrees C. The enzyme exclusively catalyzed the phosphorylation of D-glucose using ATP as a phosphoryl donor. ITP was accepted in addition to ATP. The rate dependence with both glucose and ATP followed Michaelis-Menten kinetics, with apparent K(m) values of 0.054 and 0.50 mM, respectively. The enzyme activity required divalent cations; Mg(2+), which was most effective, could partially be replaced by Mn(2+) or Ca(2+). Phylogenetic analysis revealed that the glucokinase from A. pernix does not belong to the clusters of enzymes found in bacteria and eukarya. This is the first description of the characteristics of an ATP-dependent glucokinase from an archaeon. PMID- 12761186 TI - Catalytic efficiency and some structural properties of cold-active protein tyrosine-phosphatase. AB - A procedure was established for expression and purification of abundant recombinant cold-active protein-tyrosine-phosphatase (RCPTPase), which showed identical enzymatic characteristics to the native enzyme (NCPTPase). The purified RCPTPase showed high catalytic activity at low temperature and maximal activity at 30 degrees C. RCPTPase has a thermodynamic characteristic in that its activation enthalpy was determined to be low, 4.3 kcal/mol, at temperatures below 19.3 degrees C, where the Arrhenius relationship exhibited an inflection point, in comparison with 20.3 kcal/mol above 19.3 degrees C. Also, the thermostability, DeltaG(water), of the catalytic site in the RCPTPase molecule was increased with a decrease in temperature. It was considered that cold-active protein-tyrosine phosphatase could maintain its catalytic site in a stable conformation for eliciting high catalytic activity with low activation enthalpy at low temperature. PMID- 12761187 TI - Characterization of complement C3 as a glycyrrhizin (GL)-binding protein and the phosphorylation of C3alpha by CK-2, which is potently inhibited by GL and glycyrrhetinic acid in vitro. AB - The physiological interaction between glycyrrhizin (GL) and serum complement C3, and the inhibitory effects of GL, glycyrrhetinic acid (GA), and a GA derivative (oGA) on the phosphorylation of C3 by casein kinase 2 (CK-2), were investigated in vitro. C3 was found to be a GL-binding protein (gbP), because (i) of its high affinity for a GL-affinity HPLC column; and (ii) both GL and GA induce conformational changes in C3. At least four trypsin-resistant fragments (p30, p25, p18, and p15) were detected when the (32)P-labeled C3alpha was digested with trypsin in the presence of 100 micro M GA. Two of these (p25 and p15) were immuno precipitated with anti-C3a serum. Furthermore, it was found that C3a contains GL binding domains, because (i) C3a (anaphylatoxin) could be selectively purified from the synovial fluids of patients with rheumatoid arthritis by GL-affinity column chromatography (HPLC); and (ii) purified human C3a has a high affinity for a GL-affinity column. In addition, C3alpha (p115) of C3 was effectively phosphorylated by CK-2 in the presence of poly-Arg (a CK-2 activator) in vitro. This phosphorylation was completely inhibited by 10 micro M oGA, 30 micro M GA, or 100 micro M GL. Taken together, these results suggest that the GL-induced inhibition of the physiological activities of C3a and C3alpha may be involved in the anti-inflammatory effect of GL in vivo. PMID- 12761188 TI - Molecular structural and functional characterization of tumor suppressive anti ErbB-2 monoclonal antibody by phage display system. AB - To investigate the molecular structural and functional characteristics of tumor suppressive anti-ErbB-2 monoclonal antibody (mAb) SER4, we performed mAb-gene cloning and epitope mapping by a phage display system. Structural analysis demonstrated that both the heavy chain (HC) and light chain variable regions are highly homologous with the derived germline sequences, while the HC complementarity determining region (HCDR) 3 has a relatively short length and biased amino acid usage. A cloned gene-derived recombinant Fab (rFab) fragment showed antigen binding activity and specificity comparable to the parent mAb. Cross-linking of the rFab fragment with the anti-Fab antibody elicited cell growth inhibition in vitro. These results imply that the cloned genes actually encode the Fab part of SER4. The epitope mimetic peptide (mimotope) isolated by panning a phage-displayed random peptide library against SER4 showed no cross reactivity with mAbs other than SER4. The mimotope was found to be homologous with (87)AHNQVRQVPLQR(98) in the extracellular domain of ErbB-2 by means of a clustalw search. Since SER4 causes the growth inhibition of ErbB-2 positive cells, the predicted epitope sequence may constitute the putative functional domain of ErbB-2. PMID- 12761189 TI - Integration of cytochrome b5 into endoplasmic reticulum membrane: participation of carboxy-terminal portion of the transmembrane domain. AB - Integration of cytochrome b(5) (b5), a tail-anchored protein located in the endoplasmic reticulum (ER) membrane, into the membrane was studied. Mutation of three amino acids, -Leu-Met-Tyr, at the carboxy-terminal end of the transmembrane segment of b5 to alanines resulted in localization of the mutated protein, b5LMY/AAA, in the cytosol as well as in the ER membrane. When an N-glycosylation site was introduced at the carboxy-terminal end of b5LMY/AAA, a substantial amount of the glycosylated form of the mutant protein was recovered in the cytosol fraction. A portion of the mutant protein recovered in the ER was released from the membrane by incubation with the cytosol fraction, but no further release was observed in the second incubation, suggesting that b5 is present in two different states, loosely-bound and firmly-integrated forms, in the ER membrane. These results suggest that b5 is integrated into the ER membrane via the loosely bound state, in which the carboxy-terminal end of the molecule is inserted into the luminal side of the vesicle but is easily translocated back to the cytosol, and that the three amino acids are important for conversion of the loosely-bound state to the firmly-integrated state. PMID- 12761190 TI - Kinetic studies on the hydrolysis of N-acetylated and N-deacetylated derivatives of 4-methylumbelliferyl chitobioside by the family 18 chitinases ChiA and ChiB from Serratia marcescens. AB - Kinetic analyses of the hydrolysis reactions of N-acetylated and N-deacetylated derivatives of 4-methylumbelliferyl chitobioside [(GlcNAc)(2)-UMB (1), GlcN GlcNAc-UMB (2), GlcNAc-GlcN-UMB (3), and (GlcN)(2)-UMB (4)] by ChiA and ChiB from Serratia marcescens were performed. Both enzymes released UMB from all compounds apart from 4. The S-v curves of the hydrolyses of 1 by ChiA and ChiB both exhibited atypical kinetic patterns, and the shapes of the two S-v curves were different from one another. However, both curve shapes were explained by assuming some of the enzyme present formed complexes with multiple molecules of the substrate. Conversely, the S-v curves generated in the cleavage of 2 and 3 by ChiA exhibited typical Michaelis-Menten profiles. Both enzymes hydrolysed 2 with an approximately 14-fold higher K(m) value relative to 1, indicating that the N acetyl group was recognised at the -2 subsite. The k(cat) value obtained with ChiA was identical to the k(cat) value observed for 1. However, the k(cat) value for ChiB was one-fourth that of 1, suggesting that the removal of the N-acetyl group caused an increase in the formation of a non-productive ES-complex. ChiA and ChiB hydrolysed 3 with 5- and 20-fold greater K(m) values relative to 1, respectively, and 60- and 30-fold smaller k(cat) values relative to 1, respectively. The reaction mechanism of family 18 chitinases is discussed based upon the results obtained from the hydrolysis of these compounds. PMID- 12761191 TI - Differential roles of human monoamine (M)-form and simple phenol (P)-form phenol sulfotransferases in drug metabolism. AB - Cytosolic sulfotransferases (STs) are traditionally known as Phase II drug metabolizing or detoxifying enzymes that facilitate the removal of drugs and other xenobiotic compounds. In this study, we carried out a systematic investigation on the sulfation of drug compounds by two major human phenol STs (PSTs), the monoamine (M)-form and simple phenol (P)-form PSTs. Activity data obtained showed the differential substrate specificity of the two enzymes for the thirteen drug compounds tested. Kinetic studies revealed that the M-form PST displayed stereoselectivity for the chiral drug, isoproterenol. The effects of divalent metal cations on the activity of the M-form and P-form PSTs toward representative drug compounds were quantitatively evaluated. Results obtained indicated that the drug-sulfating activities of the two human PSTs were partially or completely inhibited or stimulated by the ten divalent metal cations tested at a 5 mM concentration. The two enzymes appeared to be less sensitive to the effects of physiologically more abundant metal cations such as Mg(2+) and Ca(2+), but more sensitive to the detrimental effects of other metal cations that may enter the body as environmental contaminants. PMID- 12761192 TI - Protein kinase Czeta (PKCzeta): activation mechanisms and cellular functions. AB - The zeta isotype of protein kinase C (PKCzeta) is a member of the atypical PKC subfamily and has been widely implicated in the regulation of cellular functions. Increasing evidence from studies using in vitro and in vivo systems points to PKCzeta as a key regulator of critical intracellular signaling pathways induced by various extracellular stimuli. The major activation pathway of PKCzeta depends on phosphatidylinositol (PI)-3,4,5-trisphosphate (PIP(3)), which is mainly produced by PI-3 kinase. 3'-PI-dependent protein kinase 1, which binds with high affinity to PIP(3), phosphorylates and activates PKCzeta. Many studies demonstrated the involvement of PKCzeta in the mitogen-activated protein kinase cascade, transcriptional factor NFkappaB activation, ribosomal S6-protein kinase signaling, and cell polarity. An important molecular event in a cell is the association of PKCzeta with other signaling molecules, as well as scaffold proteins, to form large complexes that regulate their pathways. The understanding of the mechanisms underlying PKCzeta-mediated control of intracellular signaling is beginning to provide important insights into the roles of PKCzeta in various cells. PMID- 12761193 TI - Protein kinase C lambda/iota (PKClambda/iota): a PKC isotype essential for the development of multicellular organisms. AB - PKClambda/iota belongs to the third group of the PKC family, atypical PKC (aPKC), together with PKCzeta based on its sequence divergence from conventional and novel PKCs observed not only in the N-terminal regulatory domain but also in the kinase domain. Although one of the most distinct features of aPKC is its single, unrepeated cysteine-rich domain, recent studies have revealed that the N-terminal regulatory domain has additional aPKC-specific structural motifs involved in various protein-protein interactions, which are important for the regulation and the subcellular targeting of aPKC. The identification of aPKC-specific binding proteins has significantly facilitated our understanding of the activation mechanism as well as the physiological function of aPKC at the molecular level. In particular, the finding that the mammalian homologs of the Caenorhabditis elegans proteins, PAR-3 and PAR-6, bind aPKC unexpectedly opens a new avenue for exploring a thus far completely unrecognized critical function of aPKC, that is, as a component of an evolutionarily conserved cell polarity machinery. Together with the great progress in the genome project as well as in the genetic analysis of model organisms, these advances are leading us into the new era of aPKC study in which functional divergence between PKClambda/iota and zeta can be discussed in elaborately. PMID- 12761194 TI - The structure and function of PKN, a protein kinase having a catalytic domain homologous to that of PKC. AB - PKN is a serine/threonine protein kinase that has a catalytic domain homologous to protein kinase C (PKC) family members and a unique regulatory region containing antiparallel coiled-coil (ACC) domains. PKN is the first identified serine/threonine protein kinase that can bind to and be activated by a small GTPase Rho, and it can also be activated by fatty acids such as arachidonic acid in vitro. PKN is widely distributed in various organisms such as mammal, frog, fly, and starfish. There are at least three different isoforms of PKN (PKNalpha/PAK-1/PRK-1, PKNbeta, and PRK2/PAK-2/PKNgamma) in mammals, each of which shows different enzymological properties, tissue distribution, and varied functions. PMID- 12761195 TI - Cloning, expression, crystallization, and preliminary X-ray analysis of recombinant mouse lipocalin-type prostaglandin D synthase, a somnogen-producing enzyme. AB - Lipocalin-type prostaglandin D synthase is the key enzyme for the production of prostaglandin D(2), a potent endogenous somnogen, in the brain. We cloned, produced, and crystallized the native enzyme and selenomethionyl Cys(65)Ala mutants of the recombinant mouse protein by the hanging drop vapor-diffusion method with both malonate and citrate as precipitants. The native crystals obtained with malonate belong to orthorhombic space group P2(1)2(1)2(1) with lattice constants a = 46.2, b = 66.8, and c = 105.3 A. The selenomethionyl crystals obtained with citrate belong to orthorhombic space group C222(1) with lattice constants a = 45.5, b = 66.8, and c = 104.5 A. The native crystals diffracted beyond 2.1 A resolution. PMID- 12761196 TI - D-arginase of Arthrobacter sp. KUJ 8602: characterization and its identity with Zn(2+)-guanidinobutyrase. AB - D-Arginase activity was found in the cells of an isolate, Arthrobacter sp. KUJ 8602, grown in the L-arginine medium, and the enzyme was purified and characterized. Its molecular weight was estimated to be about 232,000 by gel filtration, and that of the subunit was approximately 40,000 by SDS-PAGE, suggesting that the enzyme is a homohexamer. The enzyme acted on not only D arginine but also 4-guanidinobutyrate, 3-guanidinopropionate and even L-arginine. The V(max)/K(m) values for 4-guanidinobutyrate and D-arginine were determined to be 87 and 0.81 micro mol/min/mg/mM, respectively. Accordingly, the enzyme is regarded as a kind of guanidinobutyrase [EC 3.5.3.7]. The pH optima for 4 guanidinobutyrate and D-arginine were 9.0 and 9.5, respectively. The enzyme was inhibited competitively by 5-aminovalerate, and thiol carboxylates such as mercaptoacetate served as strong mixed-type inhibitors. The enzyme contained about 1 g-atom of firmly bound Zn(2+) per mol of subunit, and removal of the metal ions by incubation with 1,10-phenanthroline resulted in loss of activity. The inactivated enzyme was reactivated markedly by incubation with either Zn(2+) or Co(2+), and slightly by incubation with Mn(2+). The nucleotide sequence of enzyme contains an open reading frame that encodes a polypeptide of 353 amino acid residues (M(r): 37,933). The predicted amino acid sequence contains sequences involved in the binding of metal ions and the guanidino group of the substrate, which show a high homology with corresponding sequences of Mn(2+) dependent amidinohydrolases such as agmatinase from Escherichia coli and L arginase from rat liver, though the homology of their entire sequences is relatively low (24-43%). PMID- 12761197 TI - Human Fc epsilon RIalpha-specific human single-chain Fv (scFv) antibody with antagonistic activity toward IgE/Fc epsilon RIalpha-binding. AB - The alpha-chain of Fc epsilon RI (Fc epsilon RIalpha) plays a critical role in the binding of IgE to Fc epsilon RI. A fully human antibody interfering with this interaction may be useful for the prevention of IgE-mediated allergic diseases. Here, we describe the successful isolation of a human single-chain Fv antibody specific to human Fc epsilon RIalpha using human antibody phage display libraries. Using the non-immune phage antibody libraries constructed from peripheral blood lymphocyte cDNA from 20 healthy subjects, we isolated three phage clones (designated as FcR epsilon 27, FcR epsilon 51, and FcR epsilon 70) through two rounds of biopanning selection. The purified soluble scFv, FcR epsilon 51, inhibited the binding of IgE to recombinant Fc epsilon RIalpha, although both FcR epsilon 27 and FcR epsilon 70 showed fine binding specificity to Fc epsilon RIalpha. Since FcR epsilon 51 was determined to be a monomer by HPLC, BIAcore analysis was performed. The dissociation constant of FcR epsilon 51 to Fc epsilon RIalpha was estimated to be 20 nM, i.e., fortyfold lower than that of IgE binding to Fc epsilon RIalpha (K(d) = 0.5 nM). With these characteristics, FcR epsilon 51 exhibited inhibitory activity on the release of histamine from passively sensitized human peripheral blood mononuclear cells. PMID- 12761198 TI - Thermostable aspartase from a marine psychrophile, Cytophaga sp. KUC-1: molecular characterization and primary structure. AB - We found that a psychrophilic bacterium isolated from Antarctic seawater, Cytophaga sp. KUC-1, abundantly produces aspartase [EC4.3.1.1], and the enzyme was purified to homogeneity. The molecular weight of the enzyme was estimated to be 192,000, and that of the subunit was determined to be 51,000: the enzyme is a homotetramer. L-Aspartate was the exclusive substrate. The optimum pH in the absence and presence of magnesium ions was determined to be pH 7.5 and 8.5, respectively. The enzyme was activated cooperatively by the presence of L aspartate and by magnesium ions at neutral and alkaline pHs. In the deamination reaction, the K(m) value for L-aspartate was 1.09 mM at pH 7.0, and the S(1/2) value was 2.13 mM at pH 8.5. The V(max) value were 99.2 U/mg at pH 7.0 and 326 U/mg at pH 8.5. In the amination reaction, the K(m) values for fumarate and ammonium were 0.797 and 25.2 mM, respectively, and V(max) was 604 U/mg. The optimum temperature of the enzyme was 55 degrees C. The enzyme showed higher pH and thermal stabilities than that from mesophile: the enzyme was stable in the pH range of 4.5-10.5, and about 80% of its activity remained after incubation at 50 degrees C for 60 min. The gene encoding the enzyme was cloned into Escherichia coli, and its nucleotides were sequenced. The gene consisted of an open reading frame of 1,410-bp encoding a protein of 469 amino acid residues. The amino acid sequence of the enzyme showed a high degree of identity to those of other aspartases, although these enzymes show different thermostabilities. PMID- 12761199 TI - Development and characterization of a novel fusion protein composed of a human IgG1 heavy chain constant region and a single-chain fragment variable antibody against Venezuelan equine encephalitis virus. AB - Murine monoclonal antibody 1A4A1 has been shown to recognize a conserved neutralizing epitope of envelope glycoprotein E2 of Venezuelan equine encephalitis virus. It is a potential candidate for development of a second generation antibody for both immunodiagnosis and immunotherapy. In order to minimize the immunogenicity of murine antibodies and to confer human immune effector functions on murine antibodies, a recombinant gene fusion was constructed. It encoded a human IgG1 heavy chain constant region and a single chain fragment variable antibody of 1A4A1. After expression in bacteria as inclusion bodies, the recombinant antibody was purified and refolded in vitro. The recombinant soluble antibody was demonstrated to retain high antigen-binding affinity to Venezuelan equine encephalitis virus and to possess some human IgG crystallizable fragment domain functions, such as recognition by protein G and human complement C1q binding. On non-reducing and reducing gel electrophoresis analysis of proteolytic fragments of the recombinant antibody, disulfide bond formation was found in the hinge region of the antibody. From these data, it was concluded that the recombinant antibody was capable of antigen recognition, and retained several functional activities. This work forms the basis for characterization of the recombinant antibody as to efficacy in vivo. PMID- 12761200 TI - Characterization of novel acetyltransferases found in budding and fission yeasts that detoxify a proline analogue, azetidine-2-carboxylic acid. AB - We recently found that budding yeast Saccharomyces cerevisiae sigma1278b, but not genome project strain S288C, has a gene conferring resistance to L-azetidine-2 carboxylic acid (AZC), a toxic four-membered ring analogue of L-proline. Also, the gene, designated as MPR1, encodes a novel acetyltransferase that detoxifies AZC via acetylation. We now report the results of subsequent work. On a homology search with MPR1, we detected a gene in fission yeast Schizosaccharomyces pombe. This gene, designated as ppr1(+) (pombe MPR1), is responsible for the AZC resistance of S. pombe as judged from the results of gene disruption and overexpression experiments. Escherichia coli cells expressing ppr1(+), like ones expressing MPR1, were resistant to AZC and produced an AZC acetyltransferase. We further found that the enzymes encoded by MPR1 and ppr1(+) were homodimers, and catalyzed the acetylation of AZC but not any other L-proline-related compounds. Ppr1p was more thermostable than Mpr1p, although Ppr1p had a lower optimum temperature than Mpr1p. The higher AZC acetylation activity of Mpr1p, in comparison to that of Ppr1p, was attributed to the larger k(cat)/K(m) value for acetyl-CoA of Mpr1p than that of Ppr1p. PMID- 12761201 TI - Crystal structure of hyperthermophilic archaeal initiation factor 5A: a homologue of eukaryotic initiation factor 5A (eIF-5A). AB - Eukaryotic initiation factor 5A (eIF-5A) is ubiquitous in eukaryotes and archaebacteria and is essential for cell proliferation and survival. The crystal structure of the eIF-5A homologue (PhoIF-5A) from a hyperthermophilic archaebacterium Pyrococcus horikoshii OT3 was determined at 2.0 A resolution by the molecular replacement method. PhoIF-5A is predominantly composed of beta strands comprising two distinct folding domains, an N-domain (residues 1-69) and a C-domain (residues 72-138), connected by a short linker peptide (residues 70 71). The N-domain has an SH3-like barrel, while the C-domain folds in an (oligonucleotide/oligosaccharide binding) OB fold. Comparison of the structure of PhoIF-5A with those of archaeal homologues from Methanococcus jannaschii and Pyrobaculum aerophilum showed that the N-domains could be superimposed with root mean square deviation (rmsd) values of 0.679 and 0.624 A, while the C-domains gave higher values of 1.824 and 1.329 A, respectively. Several lines of evidence suggest that eIF-5A functions as a biomodular protein capable of interacting with protein and nucleic acid. The surface representation of electrostatic potential shows that PhoIF-5A has a concave surface with positively charged residues between the N- and C-domains. In addition, a flexible long hairpin loop, L1 (residues 33-41), with a hypusine modification site is positively charged, protruding from the N-domain. In contrast, the opposite side of the concave surface at the C-domain is mostly negatively charged. These findings led to the speculation that the concave surface and loop L1 at the N-domain may be involved in RNA binding, while the opposite side of the concave surface in the C-domain may be involved in protein interaction. PMID- 12761202 TI - Interconversion of the product specificity of type I eubacterial farnesyl diphosphate synthase and geranylgeranyl diphosphate synthase through one amino acid substitution. AB - Prenyltransferases catalyze the sequential condensation of isopentenyl diphosphate into prenyl diphosphates with specific chain lengths. Pioneering studies demonstrated that the product specificities of type I prenyltransferases were mainly determined by the amino acid residues at the 4th and 5th positions before the first aspartate-rich motif (FARM) of the prenyltransferases. We previously cloned a type I geranylgeranyl diphosphate synthase (GGDPSase) gene from Streptomyces griseolosporeus MF730-N6 [Hamano, Y., Dairi, T., Yamamoto, M., Kawasaki, T., Kaneda, K., Kuzuyama, T., Itoh, N., and Seto, H. (2001) BIOSCI: Biotechnol. Biochem. 65, 1627-1635]. In this study, a prenyltransferase gene was cloned from Streptomyces argenteolus A-2 and was confirmed to encode a type I farnesyl diphosphate synthase (FDPSase). Interestingly, the amino acid residues at the 4th and 5th positions before the FARM were the same in these two enzymes. To identify the amino acid that determines the product chain length, mutated enzymes, GGDPSase (L-50S), FDPSase (S-50L), GGDPSase (V-8A), FDPSase (A-8V), GGDPSase (A+57L), and FDPSase (L+58A), in which the amino acid residue at the 50th, -8th, and +57th (58th) position before or after the FARM was substituted with the corresponding amino acid of the other enzyme, were constructed. The GGDPSase (A+57L) and FDPSase (L+58A) produced farnesyl diphosphate and geranylgeranyl diphosphate, respectively. On the other hand, the other mutated enzymes produced prenyl diphosphates with the same chain lengths as the wild type enzymes did. These results showed that the amino acid residue at the 57th (58th) position after the FARM also played an important role in determination of the product specificity. PMID- 12761203 TI - Formation of the molten globule-like state of cytochrome c induced by n-alkyl sulfates at low concentrations. AB - The molten globule state of cytochrome c is the major intermediate of protein folding. Elucidation of the thermodynamic mechanism of conformational stability of the molten globule state would enhance our understanding of protein folding. The formation of the molten globule state of cytochrome c was induced by n-alkyl sulfates including sodium octyl sulfate, SOS; sodium decyl sulfate, SDeS; sodium dodecyl sulfate, SDS; and sodium tetradecyl sulfate, STS, at low concentrations. The refolding states of the protein were monitored by spectroscopic techniques including circular dichroism (CD), visible absorbance and fluorescence. The effect of n-alkyl sulfates on the structure of acid-unfolded horse cytochrome c at pH 2 was utilized to investigate the contribution of hydrophobic interactions to the stability of the molten globule state. The addition of n-alkyl sulfates to the unfolded state of cytochrome c appears to support the stabilized form of the molten globule. The m-values of the refolded state of cytochrome c by SOS, SDeS, SDS, and STS showed substantial variation. The enhancement of m-values as the stability criterion of the molten globule state corresponded with increasing chain length of the cited n-alkyl sulfates. The compaction of the molten globule state induced by SDS, as a prototype for other n-alkyl sulfates, relative to the unfolded state of cytochrome c was confirmed by Stokes radius and thermal transition point (T(m)) measured by microviscometry and differential scanning calorimetry (DSC), respectively. Thus, hydrophobic interactions play an important role in stabilizing the molten globule state. PMID- 12761204 TI - Inhibition of mitogen-activated kinase kinase kinase 3 activity through phosphorylation by the serum- and glucocorticoid-induced kinase 1. AB - The mitogen-activated protein kinase kinase kinase 3 (MEKK3) is a member of the MAP kinase family whose cellular activity is elevated in response to growth factors, oxidative stress, and hyperosmolar conditions. MEKK3 regulates MKK3 and MKK5/6/7. MEKK3 is involved distinctively in the signal pathway for blocking cell proliferation and cell cycle progression, contradictory to the biological responses commonly associated with other members of MEKKs. Based information concerning the substrate specificity of serum- and glucocorticoid-induced kinase 1 (SGK1), R-x-R-x-x-(S/T)-phi, where phi indicates a hydrophobic amino acid, two putative phosphorylation sites (Ser(166) and Ser(337)) were found in MEKK3. It was shown that the recombinant MEKK3 protein and fluorescein-labeled MEKK3 peptides (FITC-(159)epRsRhlSVi(168) and FITC-(330)dpRgRlpSAd(339)) are phosphorylated by SGK1 in vitro. It was also observed that the intrinsic kinase activity of MEKK3 on Ser(189) of MKK3 (equivalent to Ser(207) of MKK6) decreased along with phosphorylation of Ser(166) and Ser(337) in MEKK3 in vitro and in vivo. Therefore, it is suggested that SGK1 inhibits MEKK3-MKK3/6 signal transduction by phosphorylation of MEKK3. PMID- 12761205 TI - CIS1 interacts with the Y532 of the prolactin receptor and suppresses prolactin dependent STAT5 activation. AB - Prolactin (PRL) interacts with a single-chain prolactin-specific receptor of the cytokine receptor superfamily. PRL triggers the activation of JAK2 kinase, which phosphorylates the PRL receptor itself, and of STAT5, a member of the family of signal transducers and activators of transcription (STAT). We have shown that the STAT5-dependent immediate early gene, CIS1 (Cytokine-Inducible SH2 domain containing protein-1), suppresses PRL-induced STAT5 activation in vitro as well as in transgenic mice. To facilitate the study of the interactions between CIS1 and the PRL receptor, we have developed the yeast tri-hybrid system, a modification of the yeast two-hybrid system. We expressed CIS1 fused to the DNA binding domain and PRL receptor cytoplasmic domain fused to the transcription activation domain in the presence or absence of the tyrosine kinase domain of JAK2 in yeast. CIS1 bound to the PRL receptor cytoplasmic domain in a JAK2 dependent manner. Moreover, we determined that the phosphorylated Y532 of the murine PRL receptor is the binding site for CIS1. Interestingly, Y532 has been shown to be unnecessary for STAT5 activation, although CIS1 overexpression suppressed PRL-induced STAT5 activation. These data suggest that the suppression of STAT5 activation by CIS1 is not due to a simple competition with STAT5 but rather to a modification of the receptor by CIS1 binding. PMID- 12761206 TI - Dual subcellular distribution of cytochrome b5 in plant, cauliflower, cells. AB - Subfractionation studies showed that cytochrome b(5) (cyt b5), which has been considered to be a typical ER protein, was localized in both the endoplasmic reticulum membrane (ER) and the outer membrane of mitochondria in cauliflower (Brassica olracea) cells and was a component of antimycin A-insensitive NADH cytochrome c reductase system in both membranes. When cDNA for cauliflower cyt b5 was introduced into mammalian (COS-7) and yeast cells as well as into onion cells, the expressed cytochrome was localized both in the ER and mitochondria in those cells. On the other hand, rat and yeast cyt b5s were specifically localized in the ER membranes even in the onion cells. Mutation experiments showed that cauliflower cyt b5 carries information that targets it to the ER and mitochondria within the carboxy-terminal 10 amino acids, as in the case of rat and yeast cyt b5s, and that replacement of basic amino acids in this region of cauliflower cyt b5 with neutral or acidic ones resulted in its distribution only in the ER. Together with the established findings of the importance of basic amino acids in mitochondrial targeting signals, these results suggest that charged amino acids in the carboxy-terminal portion of cyt b5 determine its location in the cell, and that the same mechanism of signal recognition and of protein transport to organelles works in mammalian, plant, and yeast cells. PMID- 12761207 TI - Expression of a synthetic gene coding for ostrich egg-white lysozyme in Pichia pastoris and its enzymatic activity. AB - To investigate the structure-function relationships of goose-type lysozyme, a gene coding for ostrich egg-white lysozyme (OEL) was designed based on the published amino acid sequence and constructed by assembling 32 chemically synthesized oligonucleotides. To obtain the recombinant OEL (rOEL), the synthetic gene was fused to the alpha-factor signal peptide in the expression vector pPIC9K and expressed in the methylotrophic yeast Pichia pastoris. The secreted protein from the transformed yeast was found to be processed at three different sites, including the correct site. The correctly processed rOEL was purified to homogeneity and shown to be indistinguishable from the authentic form in terms of circular dichroism (CD) spectrum and enzyme activity. Furthermore, the time course of the reaction catalyzed by OEL was studied using (GlcNAc)(n) (n = 5 and 6) as the substrate and compared to that of goose egg-white lysozyme (GEL) [Honda and Fukamizo (1998) BIOCHIM: Biophys. Acta 1388, 53-65]. OEL hydrolyzed (GlcNAc)(6) in an endo-splitting manner producing mainly (GlcNAc)(2), (GlcNAc)(3), and (GlcNAc)(4), and cleavage to (GlcNAc)(3) + (GlcNAc)(3) predominated over that to (GlcNAc)(2) + (GlcNAc)(4). This indicates that OEL hydrolyzes preferentially the third glycosidic linkage from the nonreducing end of (GlcNAc)(6) as in the case of GEL. The cleavage pattern seen for (GlcNAc)(5) was similar to that seen for (GlcNAc)(6). Theoretical analysis of the reaction time-course for OEL revealed that the binding free energy values for subsites B, E, and G were different between OEL and GEL, although these lysozymes were estimated to have the same type of subsite structure. PMID- 12761208 TI - A rapid purification method for human RNA polymerase II by two-step affinity chromatography. AB - The molecular dissection of transcription mechanisms is greatly facilitated by constructing and manipulating defined transcription systems in vitro. This approach requires highly purified transcription factors. A major enzyme participating in the transcription reaction is RNA polymerase II (RNAPII), which is composed of at least 12 subunits (RPB1-12). Due to its complex structure, it is difficult to prepare highly pure RNAPII by the conventional purification procedure. We transfected HeLa cells with a plasmid expressing RPB3 with a double FLAG-histidine tag on its amino-terminus. A high yielding clone was isolated and its extracts were subjected to immunoaffinity purification and then Co(2+) affinity chromatography. This resulted in a preparation of RNAPII complexes that consisted of all the core subunits, including the double-tagged RPB3 protein. Transcription reactions with oligo (dC)-tailed templates and transcription assays involving general transcription factors revealed that the double-tagged RNAPII complexes are active and functional in basal and activated transcription. Our method is superior to the conventionally used purification procedure in that the final preparation is markedly more pure (92% versus 40%), and the procedures are much less time-consuming. Thus, this two-step affinity purification method is an uncomplicated and effective method by which active and functional RNAPII can be prepared. PMID- 12761209 TI - Enhanced synthesis of poly(3-hydroxybutyrate) in recombinant Escherichia coli by means of error-prone PCR mutagenesis, saturation mutagenesis, and in vitro recombination of the type II polyhydroxyalkanoate synthase gene. AB - Type II synthase (PhaC1(Ps)) for polyhydroxyalkanoate (PHA) from Pseudomonas sp. 61-3 was subjected to an in vitro evolution system including PCR-mediated mutagenesis in order to improve the function of PhaC1(Ps) in terms of its ability to produce poly(3-hydroxybutyrate) [P(3HB)] in recombinant Escherichia coli. Based on our established in vivo assay system, two positions (Ser325 and Gln481) where mutations provided remarkable increases in P(3HB) synthesis were identified. Saturation mutagenesis at these positions was carried out to explore whether there might be more beneficial sequences for P(3HB) synthesis than those identified in the point mutation library. As a result, five single mutants [S325C (T) and Q481M (K, R)] gave rise to highly enhanced P(3HB) synthesis. Drastically enhanced P(3HB) synthesis (up to 340- to 400-fold the amount of that of the wild type) was further achieved by generation of all five variants of the double mutants combining the codons for residues 325/481. It is feasible that the replacement of Ser (specific for type II synthase) by Thr (specific for type I synthase) at position 325 resulted in acquiring greater P(3HB) synthesis ability as exhibited by type I synthases. The other hot spot, 481, that positively contributes to enhanced P(3HB) synthesis is located adjacent to a His479, a residue that forms a putative catalytic diad that can be inferred by sequence alignment. PMID- 12761210 TI - Synthesis and structural characterization of silk-like materials incorporated with an elastic motif. AB - Genetic engineering strategies were applied to synthesize silk-like materials, [(GVPGV)(2)GG(GAGAGS)(3)AS](n). The primary structure of these materials represents the repetitive crystalline region of Bombyx mori silk fibroins incorporated with an elastic motif selected from animal elastin. The oligonucleotides were designed to encode the desired recombinant proteins and then expressed in the Escherichi coli system. The expression and purification conditions for the production of the recombinant proteins were optimized. (13)C CP/MAS NMR was used for structural characterization in the solid state, where the isotope labeling was performed using a modified M9 medium. The secondary structures of these materials are primarily governed by the designated amino acid sequence, where the B. mori silk fibroin block, (GAGAGS)(3), tends to form the crystalline region, which is interrupted by the flexible (GVPGV)(2) block. The CD data suggested that the structure of these materials was length-dependent in the solution state, i.e., a higher molecule weight leads to a higher ordered structure. PMID- 12761211 TI - Activation of TRPV1 by the satiety factor oleoylethanolamide. AB - The fatty acid oleoylethanolamide (OEA) is a satiety factor that excites peripheral vagal sensory nerves, but the mechanism by which this occurs and the molecular targets of OEA are unclear. In this study the ability of OEA to modulate the capsaicin receptor (TRPV1) was explored. OEA alone did not activate TRPV1 expressed in Xenopus oocytes under control conditions, but produced a differential modulation of agonist-evoked responses. OEA enhanced proton-gated TRPV1 currents, inhibited anandamide-evoked currents and had no effect on capsaicin-evoked responses. Following stimulation of protein kinase C (PKC), OEA alone directly activated TRPV1 channel with an EC50 of approximately 2 microm at room temperature. This effect was due to direct phosphorylation of TRPV1 because no responses to OEA were observed with mutant channels lacking critical PKC phosphorylation sites, S502A/S800A. In sensory neurons, OEA-induced Ca2+ rises that were selective for capsaicin-sensitive cells, inhibited by the TRPV1 blocker, capsazepine, and occurred in a PKC-dependent manner. Further, after PKC stimulation, OEA activated TRPV1 channels in cell-free patches suggesting a direct mode of action. Thus, TRPV1 represents a potential target for OEA and may contribute to the excitatory action of OEA on sensory nerves. PMID- 12761212 TI - Functional characterization of ERp18, a new endoplasmic reticulum-located thioredoxin superfamily member. AB - Native disulfide bond formation in the endoplasmic reticulum is a critical process in the maturation of many secreted and outer membrane proteins. Although a large number of proteins have been implicated in this process, it is clear that our current understanding is far from complete. Here we describe the functional characterization of a new 18-kDa protein (ERp18) related to protein-disulfide isomerase. We show that ERp18 is located in the endoplasmic reticulum and that it contains a single catalytic domain with an unusual CGAC active site motif and a probable insertion between beta3 and alpha3 of the thioredoxin fold. From circular dichroism and NMR measurements, ERp18 is well structured and undergoes only a minor conformational change upon dithioldisulfide exchange in the active site. Guanidinium chloride denaturation curves indicate that the reduced form of the protein is more stable than the oxidized form, suggesting that it is involved in disulfide bond formation. Furthermore, in vitro ERp18 possesses significant peptide thiol-disulfide oxidase activity, which is dependent on the presence of both active site cysteine residues. This activity differs from that of the human PDI family in that under standard assay conditions it is limited by substrate oxidation and not by enzyme reoxidation. A putative physiological role for Erp18 in native disulfide bond formation is discussed. PMID- 12761213 TI - Human kininogen gene is transactivated by the farnesoid X receptor. AB - Human kininogen belongs to the plasma kallikreinkinin system. High molecular weight kininogen is the precursor for two-chain kinin-free kininogen and bradykinin. It has been shown that the two-chain kinin-free kininogen has the properties of anti-adhesion, anti-platelet aggregation, and anti-thrombosis, whereas bradykinin is a potent vasodilator and mediator of inflammation. In this study we show that the human kininogen gene is strongly up-regulated by agonists of the farnesoid X receptor (FXR), a nuclear receptor for bile acids. In primary human hepatocytes, both the endogenous FXR agonist chenodeoxycholate and synthetic FXR agonist GW4064 increased kininogen mRNA with a maximum induction of 8-10-fold. A more robust induction of kininogen expression was observed in HepG2 cells, where kininogen mRNA was increased by chenodeoxycholate or GW4064 up to 130-140-fold as shown by real time PCR. Northern blot analysis confirmed the up regulation of kininogen expression by FXR agonists. To determine whether kininogen is a direct target of FXR, we examined the sequence of the kininogen promoter and identified a highly conserved FXR response element (inverted repeat, IR-1) in the proximity of the kininogen promoter (-66/-54). FXR/RXRalpha heterodimers specifically bind to this IR-1. A construct of a minimal promoter with the luciferase reporter containing this IR-1 was transactivated by FXR. Deletion or mutation of this IR-1 abolished FXR-mediated promoter activation, indicating that this IR-1 element is responsible for the promoter transactivation by FXR. We conclude that kininogen is a novel and direct target of FXR, and bile acids may play a role in the vasodilation and anti-coagulation processes. PMID- 12761214 TI - Crystal structure of human DJ-1, a protein associated with early onset Parkinson's disease. AB - We report the crystal structure at 1.8-A resolution of human DJ-1, which has been linked to early onset Parkinson's disease. The monomer of DJ-1 contains the alpha/beta-fold that is conserved among members of the DJ-1/ThiJ/PfpI superfamily. However, the structure also contains an extra helix at the C terminus, which mediates a novel mode of dimerization for the DJ-1 proteins. A putative active site has been identified near the dimer interface, and the residues Cys-106, His-126, and Glu-18 may play important roles in the catalysis by this protein. Studies with the disease-causing L166P mutant suggest that the mutation has disrupted the C-terminal region and the dimerization of the protein. The DJ-1 proteins may function only as dimers. The Lys to Arg mutation at residue 130, the site of sumoylation of DJ-1, has minimal impact on the structure of the protein. PMID- 12761215 TI - Identification of a key determinant of ryanodine receptor type 1 required for activation by 4-chloro-m-cresol. AB - 4-Chloro-m-cresol (4-CmC) is a potent and specific activator of the intracellular Ca2+ release channel, the ryanodine receptor (RyR). We have previously shown that RyR1 expressed in dyspedic 1B5 myotubes is activated by 4-CmC, whereas RyR3 is not (Fessenden, J. D., Wang, Y., Moore, R. A., Chen, S. R. W., Allen, P. D., and Pessah, I. N. (2000) Biophys. J. 79, 2509-2525). To identify region(s) on RyR1 that are responsible for mediating activation by 4-CmC, we expressed RyR1-RyR3 chimeric proteins in dyspedic 1B5 myotubes and then measured 4-CmC-induced increases in intracellular Ca2+. Substitution of the C-terminal third of RyR1 into RyR3 imparted 4-CmC sensitivity to the resulting chimera, thus suggesting that determinants required for activation by 4-CmC are located in this region. We subdivided the C-terminal third of RyR1 into smaller segments and identified two overlapping regions of RyR1 (amino acids 3769-4180 and 4007-4382) that each imparted 4-CmC sensitivity to RyR3. Substitution of the 173 amino acids of RyR1 common to these two chimeras (amino acids 4007-4180) also weakly restored 4-CmC sensitivity in the resulting chimera. To confirm these findings, we created a complementary set of chimeras containing RyR3 substitutions in RyR1. Substitution of the RyR3 C terminus into RyR1 disrupted 4-CmC sensitivity in the resulting chimera. In addition, substitution of the corresponding RyR3 sequence into positions 4007-4180 of RyR1 disrupted 4-CmC sensitivity. Taken together, these results suggest that essential determinants required for activation of RyR1 by 4 CmC reside within a 173-amino acid region between residues 4007 and 4180. PMID- 12761216 TI - The mechanism of Mycobacterium tuberculosis alkylhydroperoxidase AhpD as defined by mutagenesis, crystallography, and kinetics. AB - AhpD, a protein with two cysteine residues, is required for physiological reduction of the Mycobacterium tuberculosis alkylhydroperoxidase AhpC. AhpD also has an alkylhydroperoxidase activity of its own. The AhpC/AhpD system provides critical antioxidant protection, particularly in the absence of the catalase peroxidase KatG, which is suppressed in most isoniazid-resistant strains. Based on the crystal structure, we proposed recently a catalytic mechanism for AhpD involving a proton relay in which the Glu118 carboxylate group, via His137 and a water molecule, deprotonates the catalytic residue Cys133 (Nunn, C. M., Djordjevic, S., Hillas, P. J., Nishida, C., and Ortiz de Montellano, P. R. (2002) J. Biol. Chem. 277, 20033-20040). A possible role for His132 in subsequent formation of the Cys133-Cys130 disulfide bond was also noted. To test this proposed mechanism, we have expressed the H137F, H137Q, H132F, H132Q, E118F, E118Q, C133S, and C130S mutants of AhpD, determined the crystal structures of the H137F and H132Q mutants, estimated the pKa values of the cysteine residues, and defined the kinetic properties of the mutant proteins. The collective results strongly support the proposed catalytic mechanism for AhpD. PMID- 12761217 TI - The forkhead transcription factor FOXO4 induces the down-regulation of hypoxia inducible factor 1 alpha by a von Hippel-Lindau protein-independent mechanism. AB - Tumors utilize hyperactivation of the phosphatidylinositol 3-kinase (PI3K)/AKT signaling pathway to cope with deleterious environmental conditions. Activation of the PI3K/AKT pathway has been shown to increase protein expression of the alpha subunit of the hypoxia-inducible factor (HIF) 1, a key regulator of oxygen homeostasis. Elevated levels of HIF-1 alpha induce expression of genes with critical roles in angiogenesis, erythropoiesis, and glucose metabolism, processes that are essential for tumor expansion. Here we examine the involvement of FOXO4 (also known as AFX), a member of the forkhead transcription factor superfamily that is negatively regulated by the PI3K/AKT pathway, in the regulation of HIF-1 alpha protein expression. Nuclear expression of FOXO4 results in the suppression of various responses to hypoxia, including decreased vascular endothelial growth factor, glucose transporter 1, and erythropoietin expression. Interestingly, FOXO4 down-regulates the HIF-1 alpha protein levels, consistent with the lack of hypoxia responsiveness. Previous results have revealed a role for prolyl hydroxylation and resultant von Hippel-Lindau protein (pVHL) interactions in the ubiquitin-proteasome-mediated degradation of HIF-1 alpha. However, neither inhibition of prolyl hydroxylases nor mutation of HIF-1 alpha-hydroxylated prolines involved with pVHL-mediated binding inhibits the observed FOXO4-mediated down-regulation of HIF-1 alpha. These results suggest a novel alternate mechanism for hypoxic regulation that is dependent upon the level of activation of FOXO4 mediated transcription. PMID- 12761218 TI - The Pleckstrin homology domains of phospholipases C-beta and -delta confer activation through a common site. AB - Mammalian inositol-specific phospholipase C-beta2 (PLC beta 2) and PLC delta 1 differ in their cellular activators. PLC beta 2 can be activated by G beta gamma subunits, whereas PLC delta 1 can be activated by phosphatidylinositol 4,5 bisphosphate (PI(4,5)P2). For both proteins, the N-terminal pleckstrin homology (PH) domain appears to mediate activation. Here, we have constructed a chimera in which we placed the N-terminal PH domain of PLC delta 1 into remaining C-terminal regions of PLC beta 2. The PH delta PLC beta chimera showed PI(4,5)P2-dependent membrane binding similar to PLC delta 1 and a G beta gamma interaction energy close to that of PLC delta 1. Like PLC delta 1, the chimera was activated by PI(4,5)P2 through the PH domain but not by G beta gamma. Because these and previous results indicate a common site of contact between the PH and catalytic domains in these two enzymes, we computationally docked the known structures of the PH and catalytic domains of PLC delta 1. A synthetic peptide whose sequence matches a potential interaction site between the two domains inhibited the basal activity of PLC beta 2, PLC delta 1, and a G beta gamma-activable PH beta 2-PLC delta 1 chimera. Also, the peptide was able to inhibit PI(4,5)P2 and G beta gamma activation of the PH-PLC delta 1 PH-PLC beta 2 enzymes in a concentration dependent manner, suggesting that this is the region responsible for PH domain mediated activation of the catalytic core. PMID- 12761219 TI - Candida albicans Ssa1/2p is the cell envelope binding protein for human salivary histatin 5. AB - Salivary histatins are a family of small histidine-rich peptides with potent antifungal activity. We previously identified a 70-kDa cell envelope protein in Candida albicans and Saccharomyces cerevisiae that mediates binding of histatin (Hst) 5. Isolation of Hst 5-binding protein followed by matrix-assisted laser desorption ionization mass spectrometry analysis identified this protein as the heat shock protein Ssa1p. Ssa protein and Hst 5-binding protein were found to be co-localized on immunoblots of yeast beta-mercaptoethanol cell wall extracts and cytosolic fractions. Yeast two-hybrid analysis showed strong interactions between Ssa1p and both Hst 3 and Hst 5. To assess functional roles of Ssa proteins in the Hst 5 antifungal mechanism in vivo, both binding and fungicidal assays were carried out using S. cerevisiae isogenic SSA1/SSA2 mutants. 125I-Hst 5 binding assays showed saturable binding (Kd = 2.57 x 10(-6) m) with the wild-type SSA1/SSA2 strain; however, Hst 5 binding with the Deltassa1ssa2 double mutant was reduced (Kd = 1.25 x 10(-6) m). Cell wall HSP70 proteins were also diminished, but still detectable, in S. cerevisiae Deltassa1ssa2 cells and are likely to be Ssa3p or Ssa4p. Hst 5 (31 microm) killed 80% of the wild-type cells in fungicidal assays at room temperature. However, only 50-60% killing of the single mutants (Deltassa1 and Deltassa2) was observed, and fungicidal activity was further reduced to 20-30% in the Deltassa1ssa2 double mutant. Incubation of cells under heat shock conditions increased the sensitivity of cells to Hst 5, which correlated with increased Hst 5-binding activity in Deltassa1ssa2 cells, but not in wild-type cells. This study provides evidence for a novel function for yeast Ssa1/2 proteins as cell envelope binding receptors for Hst 5 that mediate fungicidal activity. PMID- 12761220 TI - Mild heat and proteotoxic stress promote unique subcellular trafficking and nucleolar accumulation of RGS6 and other RGS proteins. Role of the RGS domain in stress-induced trafficking of RGS proteins. AB - RGS proteins comprise a large family of proteins named for their ability to negatively regulate heterotrimeric G protein signaling. RGS6 is a member of the R7 subfamily of RGS proteins possessing DEP (disheveled/Egl-10/pleckstrin) homology and GGL (G protein gamma-subunit-like) domains in addition to the semiconserved RGS domain. Our previous study documented unusual complexity in splicing of the human RGS6 gene, and we demonstrated localization of various RGS6 splice forms at sites other than the plasma membrane, including the cytoplasm and nucleus, where G proteins are not localized (Chatterjee, T. K., Liu, Z., and Fisher, R. A. (2003) J. Biol. Chem. 278, 30261-30271). Here we provide new evidence that mild heat stress, proteasome-mediated proteotoxic stress, and HSF1 expression induces dramatic relocalization of RGS6 proteins from such sites to nucleoli. This response was observed in COS-7 cells expressing various splice forms of RGS6, was not elicited by other forms of cellular stress and was observed in cells treated with various protein kinase inhibitors or co-expressing a dominant-negative kinase inactive SAPK. The RGS domain of RGS6 was identified as a primary structural module providing support for its stress-induced nucleolar trafficking and various other RGS proteins or their isolated RGS domains similarly undergo nucleolar migration in response to heat or proteotoxic stress or during co-expression of HSF1. The atypical RGS domains of axin and AKAP10 also underwent stress-induced nucleolar trafficking while structural domains outside of the RGS domain of some RGS proteins can override nucleolar trafficking in response to stress. Inhibition of rDNA transcription also promoted nucleolar migration of RGS6, a response previously observed in a subset of nucleolar proteins. The DEP domain of RGS6, but not its RGS domain, conferred structural support for its transcription-linked nucleolar migration. RGS6 exhibited trafficking from subnuclear dots to nucleoli in response to heat-, proteotoxic- or transcription-linked stress. These results provide new evidence that mammalian RGS proteins undergo unique subcellular trafficking in response to specific forms of cellular stress and implicate the RGS family of proteins in cellular stress signaling pathways. PMID- 12761221 TI - Human RGS6 gene structure, complex alternative splicing, and role of N terminus and G protein gamma-subunit-like (GGL) domain in subcellular localization of RGS6 splice variants. AB - RGS proteins are defined by the presence of a semiconserved RGS domain that confers the GTPase-activating activity of these proteins toward certain G alpha subunits. RGS6 is a member of a subfamily of RGS proteins distinguished by the presence of DEP and GGL domains, the latter a G beta 5-interacting domain. Here we report identification of 36 distinct transcripts of human RGS6 that arise by unusually complex processing of the RGS6 gene, which spans 630 kilobase pairs of genomic DNA in human chromosome 14 and is interrupted by 19 introns. These transcripts arise by use of two alternative transcription sites and complex alternative splicing mechanisms and encode proteins with long or short N-terminal domains, complete or incomplete GGL domains, 7 distinct C-terminal domains and a common internal domain where the RGS domain is found. The role of structural diversity in the N-terminal and GGL domains of RGS6 splice variants in their interaction with G beta 5 and subcellular localization and of G beta 5 on RGS6 protein localization was examined in COS-7 cells expressing various RGS6 splice variant proteins. RGS6 splice variants with complete GGL domains interacted with G beta 5, irrespective of the type of N-terminal domain, while those lacking a complete GGL domain did not. RGS6 protein variants displayed subcellular distribution patterns ranging from an exclusive cytoplasmic to exclusive nuclear/nucleolar localization, and co-expression of G beta 5 promoted nuclear localization of RGS6 proteins. Analysis of our results show that the long N terminal and GGL domain sequences of RGS6 proteins function as cytoplasmic retention sequences to prevent their nuclear/nucleolar accumulation. These findings provide the first evidence for G beta 5-independent functions of the GGL domain and for a role of G beta 5 in RGS protein localization. This study reveals extraordinary complexity in processing of the human RGS6 gene and provides new insights into how structural diversity in the RGS6 protein family is involved in their localization and likely function(s) in cells. PMID- 12761222 TI - Thermodynamic dissection of a low affinity protein-protein interface involved in human immunodeficiency virus assembly. AB - Homo-dimerization of the capsid protein CA of human immunodeficiency virus through its C-terminal domain constitutes an early crucial step in the virion assembly pathway and a potential target for antiviral inhibitors. We have truncated to alanine the 20 amino acid side chains per monomer that participate in intersubunit contacts at the CA dimer interface and analyzed their individual energetic contribution to protein association and stability. About half of the side chains in the contact epitope are critically involved in the energetic epitope as their truncation essentially prevented dimerization. However, dimerization affinity is kept low partly because of the presence of interfacial side chains whose individual truncation improves affinity by 2-20-fold. Many side chains at the interface are energetically important also for the folding of a monomeric intermediate and for its conformational rearrangement during dimerization. The thermodynamic description of this low affinity interface (dissociation constant of approximately 10 microm) was compared with those obtained for the other protein-protein interfaces, nearly all of them of much higher affinity, that have been systematically analyzed by mutation. The results reveal differences that may have been evolutionary selected and that may be exploited for the design of an effective interfacial inhibitor of human immunodeficiency virus assembly. PMID- 12761223 TI - Rab5-stimulated up-regulation of the endocytic pathway increases intracellular beta-cleaved amyloid precursor protein carboxyl-terminal fragment levels and Abeta production. AB - We previously identified abnormalities of the endocytic pathway in neurons as the earliest known pathology in sporadic Alzheimer's disease (AD) and Down's syndrome brain. In this study, we modeled aspects of these AD-related endocytic changes in murine L cells by overexpressing Rab5, a positive regulator of endocytosis. Rab5 transfected cells exhibited abnormally large endosomes immunoreactive for Rab5 and early endosomal antigen 1, resembling the endosome morphology seen in affected neurons from AD brain. The levels of both Abeta40 and Abeta42 in conditioned medium were increased more than 2.5-fold following Rab5 overexpression. In Rab5 overexpressing cells, the levels of beta-cleaved amyloid precursor protein (APP) carboxyl-terminal fragments (betaCTF), the rate-limiting proteolytic intermediate in Abeta generation, were increased up to 2-fold relative to APP holoprotein levels. An increase in beta-cleaved soluble APP relative to alpha-cleaved soluble APP was also observed following Rab5 overexpression. BetaCTFs were co-localized by immunolabeling to vesicular compartments, including the early endosome and the trans-Golgi network. These results demonstrate a relationship between endosomal pathway activity, betaCTF generation, and Abeta production. Our findings in this model system suggest that the endosomal pathology seen at the earliest stage of sporadic AD may contribute to APP proteolysis along a beta-amyloidogenic pathway. PMID- 12761224 TI - Decorin-binding sites in the adhesin DbpA from Borrelia burgdorferi: a synthetic peptide approach. AB - Lyme disease is caused by the spirochete Borrelia burgdorferi following transmission from infected Ixodes ticks to human hosts. Following colonization of the skin, spirochetes can disseminate throughout the body, resulting in complications that can include ocular, cardiac, neural, and skeletal disease. We have previously shown that B. burgdorferi expresses two closely related decorin binding adhesins (DbpA and DbpB) of the MSCRAMM (microbial surface component recognizing adhesive matrix molecule) type that can mediate bacterial attachment to extracellular matrices in the host. Furthermore, three Lys residues in DbpA appear to be critical for the binding of DbpA to decorin. We have now characterized the interaction of DbpA and decorin further by using a synthetic peptide approach. We synthesized a panel of peptides that spanned the DbpA sequence and examined their ability to inhibit the binding of intact DbpA to decorin. From these studies, we identified a decorin-binding peptide that lost this activity if the sequence was either scrambled or if a critical Lys residue was chemically modified. A minimal decorin-binding peptide was identified by examining a set of truncated peptides. One peptide is proposed to contain the primary decorin-binding site in DbpA. By comparing the amino acid sequences of 29 different DbpA homologs from different B. burgdorferi sensu lato isolates, we discovered that the identified decorin-binding sequence was quite variable. Therefore, we synthesized a new panel of peptides containing the putative decorin binding sequence of the different DbpA homologs. All of these peptides were active in our decorin-binding assay, and consensus decorin binding motifs are discussed. PMID- 12761226 TI - Dose-dependent blockade to cardiomyocyte hypertrophy by histone deacetylase inhibitors. AB - Postnatal cardiac myocytes respond to stress signals by hypertrophic growth and activation of a fetal gene program. Recently, we showed that class II histone deacetylases (HDACs) suppress cardiac hypertrophy, and mice lacking the class II HDAC, HDAC9, are sensitized to hypertrophic signals. To further define the roles of HDACs in cardiac hypertrophy, we analyzed the effects of HDAC inhibitors on the responsiveness of primary cardiomyocytes to hypertrophic agonists. Paradoxically, HDAC inhibitors imposed a dose-dependent blockade to hypertrophy and fetal gene activation. We conclude that distinct HDACs play positive or negative roles in the control of cardiomyocyte hypertrophy. HDAC inhibitors are currently being tested in clinical trials as anti-cancer agents. Our results suggest that these inhibitors may also hold promising clinical value as therapeutics for cardiac hypertrophy and heart failure. PMID- 12761225 TI - Chondroitin sulfate synthase-2. Molecular cloning and characterization of a novel human glycosyltransferase homologous to chondroitin sulfate glucuronyltransferase, which has dual enzymatic activities. AB - Chondroitin sulfate is found in a variety of tissues as proteoglycans and consists of repeating disaccharide units of N-acetylgalactosamine and glucuronic acid residues with sulfate residues at various places. We found a novel human gene (GenBank accession number AB086063) that possesses a sequence homologous with the human chondroitin sulfate glucuronyltransferase gene which we recently cloned and characterized. The full-length open reading frame encodes a typical type II membrane protein comprising 775 amino acids. The protein had a domain containing beta 3-glycosyltransferase motif but lacked a typical beta 4 glycosyltransferase motif, which is the same as chondroitin sulfate glucuronyltransferase, whereas chondroitin synthase had both domains. The putative catalytic domain was expressed in COS-7 cells as a soluble enzyme. Surprisingly, both glucuronyltransferase and N-acetylgalactosaminyltransferase activities were observed when chondroitin, chondroitin sulfate, and their oligosaccharides were used as the acceptor substrates. The reaction products were identified to have the linkage of GlcUA beta 1-3GalNAc and GalNAc beta 1-4GlcUA at the non-reducing terminus of chondroitin for glucuronyltransferase activity and N-acetylgalactosaminyltransferase activity, respectively. Quantitative real time PCR analysis revealed that the transcripts were ubiquitously expressed in various human tissues but highly expressed in the pancreas, ovary, placenta, small intestine, and stomach. These results indicate that this enzyme could synthesize chondroitin sulfate chains as a chondroitin sulfate synthase that has both glucuronyltransferase and N-acetylgalactosaminyltransferase activities. Sequence analysis based on three-dimensional structure revealed the presence of not typical but significant beta 4-glycosyltransferase architecture. PMID- 12761227 TI - Sequences within domain II of the urokinase receptor critical for differential ligand recognition. AB - The receptor for urokinase-type plasminogen activator (uPAR) plays important roles in a number of physiological and pathological processes by virtue of its interactions with urokinase-type plasminogen activator (uPA), vitronectin (Vn), and several other proteins. The uPA binding site spans all three domains (D1 to D3) of uPAR. However, the nature of the Vn binding site within uPAR is still not clear. In this study, we conducted homolog-scanning mutagenesis on uPAR by switching 14 individual segments of 4-8 residues to their counterpart sequences of a uPAR homolog CD59. All 14 mutants were well expressed, reacted with a panel of monoclonal antibodies, and exhibited correct molecular weights. Of these 14 mutants, six mutants were defective in both uPA and Vn binding. Most importantly, we found two unique mutants uPAR(Asn172-Lys175) and uPAR(Glu183-Asn186) within the D2 domain, which displayed differential ligand binding activity: both had high affinity uPA binding, but completely lost Vn binding, indicating that these two sequences constitute a novel Vn binding site. Indeed, two peptides, P1 (153CPGSNGFHNNDTFHFLKC) and P2 (171CNTTKCNEGPILELENLPQ), derived from the sequences of the identified uPA and Vn binding pockets within D2, respectively, behaved like bona fide ligand binding sites: peptide P1 bound uPA but not Vn, whereas peptide P2 bound Vn and inhibited uPAR-mediated cell adhesion, but did not interact with uPA. Altogether, our data demonstrated that uPAR D2 contains two distinct ligand binding sites for uPA and Vn. Such information will help us better understand the complex roles of uPAR in cell adhesion, migration, and tumor metastasis. PMID- 12761228 TI - How hydrophobic is alanine? AB - By a number of measures, alanine is poised at the threshold between those amino acids that promote the membrane integration of transmembrane alpha-helices and those that do not. We have measured the preference of alanine to partition into the lipid-water interface region over the central acyl chain region of the endoplasmic reticulum (ER) membrane both by its ability to promote the formation of so-called helical hairpins, i.e. a pair of transmembrane helices separated by a tight turn, and by mapping the position relative to the membrane of the lumenal end of a transmembrane alpha-helix that ends with a block of 10 alanines. Both measures show that Ala has a weak but distinct preference for the interface region, which is in agreement with recent biophysical measurements on pentaeptide partitioning in simple water-lipid or water-octanol systems (Jayasinghe, S., Hristova, K., and White, S. H. (2001) J. Mol. Biol. 312, 927-934). Considering the complexity of the translocon-mediated insertion of membrane proteins into the ER, the agreement between the biochemical and biophysical measurements is striking and suggests that protein-lipid interactions are already important during the very early steps of membrane protein assembly in the ER. PMID- 12761229 TI - ERK 1/2- and JNKs-dependent synthesis of interleukins 6 and 8 by fibroblast-like synoviocytes stimulated with protein I/II, a modulin from oral streptococci, requires focal adhesion kinase. AB - Protein I/II, a pathogen-associated molecular pattern from oral streptococci, is a potent inducer of interleukin-6 (IL-6) and IL-8 synthesis and release from fibroblast-like synoviocytes (FLSs), cells that are critically involved in joint inflammation. This synthesis implicates ERK 1/2 and JNKs as well as AP-1-binding activity and nuclear translocation of NF-kappaB. The mechanisms by which protein I/II activates MAPKs remain, however, elusive. Because focal adhesion kinase (FAK) was proposed to play a role in signaling to MAPKs, we examined its ability to contribute to the MAPKs-dependent synthesis of IL-6 and IL-8 in response to protein I/II. We used FAK-/- fibroblasts as well as FAK+/+ fibroblasts and FLSs transfected with FRNK, a dominant negative form of FAK. The results demonstrate that IL-6 and IL-8 release in response to protein I/II was strongly inhibited in both protein I/II-stimulated FAK-/- and FRNK-transfected cells. Cytochalasin D, which inhibits protein I/II-induced phosphorylation of FAK (Tyr-397), had no effect either on activation of ERK 1/2 and JNKs or on IL-6 and IL-8 release. Taken together, these results indicate that IL-6 and IL-8 release by protein I/II activated FLSs is regulated by FAK independently of Tyr-397 phosphorylation. PMID- 12761231 TI - Induced repatterning of type XVIII collagen associates with ectopic Sonic hedgehog and lung surfactant C gene expression and changes in epithelial epigenesis in the ureteric bud. AB - How cell and tissue interactions lead to complex organ structures and differentiated cell types during organogenesis is one of the most fundamental questions in developmental biology. The embryonic lung and kidney of the mouse are useful models for studying the molecular mechanisms of morphogenesis, and in both of these organs, the epithelial bud undergoes a characteristic branching process. This review discusses the potential role of an extracellular matrix molecule, type XVIII collagen, in the generation of the branching patterns in the lung and kidney and how its experimental respecification in tissue recombinants between the ureteric bud and lung mesenchyme correlates with changes in expression of signaling molecules such as sonic hedgehog and changes in cell fate as judged by ectopic expression of the lung surfactant C gene. PMID- 12761232 TI - Transcriptional control of epithelial differentiation during kidney development. PMID- 12761233 TI - Terminal differentiation of epithelia from trophectoderm to the intercalated cell: the role of hensin. AB - The intercalated cells of the collecting tubules of mammalian kidneys were discovered by Haggege and Richet to change their morphology in response to a variety of physiologic stimuli related to changes in acid base status. Recent studies showed that the conversion of beta to alpha intercalated cell under the influence of acidification of the medium is due to the deposition of hensin in the extracellular matrix of these cells and activation of a novel inductive signal transduction pathway. The conversion of beta to alpha cells is shown to be a process of terminal differentiation. Hensin is secreted as a monomer, and activation of the cell induces two activities that convert it to a dimer by folding and into a fiber by bundling of the folded dimers by galectin 3. Only the fiber is functional. Hensin is expressed in most epithelial cells, and its staining pattern suggests that it might be involved in the terminal differentiation of most epithelia. There is loss of heterozygosity of hensin in a large number of epithelial and neural tumors, making it likely that it is a tumor suppressor gene. PMID- 12761234 TI - Podocyte differentiation and hereditary proteinuria/nephrotic syndromes. AB - The study of familial nephrotic syndromes (NS) and the analysis of murine models of glomerular diseases resulted in major progresses in the knowledge of podocyte physiology and pathology. Numerous proteins participating in the composition of the slit diaphragm region have been identified. The importance of several of them (nephrin, podocin, CD2AP, and Neph1) in the maintenance of the glomerular filtration barrier has been demonstrated by the occurrence of massive proteinuria when they are defective. The role of the cytoskeleton has been revealed by the development of proteinuria/NS in patients with ACTN4 mutation and the occurrence of early and severe NS in alpha-actinin-4-deficient mice. Given the genetic heterogeneity of familial NS and the many other genes to be identified, further insights in the molecular basis of the role of the podocyte in the maintenance of the glomerular filtration barrier may be expected in the near future. PMID- 12761235 TI - Antenatal membranous glomerulonephritis with vascular injury induced by anti neutral endopeptidase antibodies: toward new concepts in the pathogenesis of glomerular diseases. PMID- 12761236 TI - Tubular shear stress and phenotype of renal proximal tubular cells. AB - Phenotypic alterations resulting from flow-induced mechanical strains is a growing field of research in many cell types such as vascular endothelial and smooth muscle cells, chondrocytes, and osteocytes. Because renal mass reduction is followed by a dramatic increase in GFR in the remaining nephron, modulation of tubular cell phenotype by flow-induced mechanical strains could be one of the events initiating the deleterious pathways that lead to the destruction of renal parenchyma after renal mass reduction. This study demonstrates that increased flow induced, in vitro and in vivo, a reinforcement of the apical domain of actin cytoskeleton and an inhibition of plasminogen activator expression. These effects of flow on plasminogen activator expression were prevented by blocking the reorganization of actin cytoskeleton and were associated with an increase in a shear-stress responsive element binding activity. These results confirm that tubular flow affects the phenotype of renal epithelial cells and suggest that flow-induced mechanical strains could be one determinant of tubulointerstitial lesions during the progression of renal diseases. PMID- 12761237 TI - Proteinuria and phenotypic change of proximal tubular cells. PMID- 12761238 TI - Transdifferentiation of epithelial glomerular cells. PMID- 12761239 TI - Bone marrow stem cells contribute to healing of the kidney. AB - A variety of recent studies support the existence of pathways, in adult humans and rodents, that allow adult stem cells to be surprisingly flexible in their differentiation repertoires. Termed plasticity, this property allows adult stem cells, assumed until now to be committed to generating a fixed range of progeny, on relocation to switch to make other specialized sets of cells appropriate to their new niche. Cells normally present within the bone marrow seem particularly flexible and are able to contribute usefully to many recipient organs. In studies of the liver, bone marrow-derived cells are seen with specialized structural and metabolic adaptations commensurate with their new locations, and these may be abundant, even sufficient, to rescue recipient mice from genetic defects and with evidence that they have proliferated in situ. In the kidney, several studies provide evidence for the presence of "reprogrammed" cells, but in most, it remains possible that cells arrive and redifferentiate but are no longer stem cells. Nevertheless, that appropriately differentiated cells are delivered deep within organs simply by injection of bone marrow cells should make us think differently about the way organs regenerate and repair. Migratory pathways for multipotential cells could be exploited to effect repairs using an individual's own stem cells, perhaps after gene therapy. This concept makes it clear that a transplanted organ would in time become affected by the genetic susceptibilities of the recipient, because of phenotypes that are expressed when trafficking cells incorporate and differentiate. PMID- 12761240 TI - Dedifferentiation and proliferation of surviving epithelial cells in acute renal failure. AB - In contrast to the heart or brain, the kidney can completely recover from an ischemic or toxic insult that results in cell death. During recovery from ischemia/reperfusion injury, surviving tubular epithelial cells dedifferentiate and proliferate, eventually replacing the irreversibly injured tubular epithelial cells and restoring tubular integrity. Repair of the kidney parallels kidney organogenesis in the high rate of DNA synthesis and apoptosis and in patterns of gene expression. As has been shown by proliferating cell nuclear antigen and 5 bromo 2'-deoxyuridine labeling studies and, in unpublished studies, by counting mitotic spindles identified by labeling with antitubulin antibody, the proliferative response is rapid and extensive, involving many of the remaining cells of the proximal tubule. This extensive proliferative capacity is interpreted to reflect the intrinsic ability of the surviving epithelial cell to adapt to the loss of adjacent cells by dedifferentiating and proliferating. Adhesion molecules likely play important roles in the regulation of renal epithelial cell migration, proliferation, and differentiation, as do cytokines and chemokines. Better understanding of all of the characteristics resulting in dedifferentiation and proliferation of the proximal tubule epithelial cell and cell-cell and cell-matrix interactions important for this repair function will lead to novel approaches to therapies designed to facilitate the processes of recovery in humans. PMID- 12761241 TI - Mutations in the human Na-K-2Cl cotransporter (NKCC2) identified in Bartter syndrome type I consistently result in nonfunctional transporters. AB - Bartter syndrome (BS) is a heterogeneous renal tubular disorder affecting Na-K-Cl reabsorption in the thick ascending limb of Henle's loop. BS type I patients typically present with profound hypokalemia and metabolic alkalosis. The main goal of the present study was to elucidate the functional implications of six homozygous mutations (G193R, A267S, G319R, A508T, del526N, and Y998X) in the bumetanide-sensitive Na-K-2Cl cotransporter (hNKCC2) identified in patients diagnosed with BS type I. To this end, capped RNA (cRNA) of FLAG-tagged hNKCC2 and the corresponding mutants was injected in Xenopus laevis oocytes and transporter activity was measured after 72 h by means of a bumetanide-sensitive (22)Na(+) uptake assay at 30 degrees C. Injection of 25 ng of hNKCC2 cRNA resulted in bumetanide-sensitive (22)Na(+) uptake of 2.5 +/- 0.5 nmol/oocyte per 30 min. Injection of 25 ng of mutant cRNA yielded no significant bumetanide sensitive (22)Na(+) uptake. Expression of wild-type and mutant transporters was confirmed by immunoblotting, showing significantly less mutant protein compared with wild-type at the same cRNA injection levels. However, when the wild-type cRNA injection level was reduced to obtain a protein expression level equal to that of the mutants, the wild-type still exhibited a significant bumetanide sensitive (22)Na(+) uptake. Immunocytochemical analysis showed immunopositive staining of hNKCC2 at the plasma membrane for wild-type and all studied mutants. In conclusion, mutations in hNKCC2 identified in type I BS patients, when expressed in Xenopus oocytes, result in a low expression of normally routed but functionally impaired transporters. These results are in line with the hypothesis that the mutations in hNKCC2 are the underlying cause of the clinical abnormalities seen in patients with type I BS. PMID- 12761243 TI - Cyclosporine A amplifies Ca2+ signaling pathway in LLC-PK1 cells through the inhibition of plasma membrane Ca2+ pump. AB - Cyclosporine A (CsA), a neutral, highly hydrophobic cyclic peptide with 11 amino acids, is currently the most widely used immunosuppressive drug for preventing graft rejection and autoimmune diseases. Despite its efficacy, the use of CsA is limited by severe side effects, mainly nephrotoxicity and arterial hypertension. Single cell microfluorimetry was used to evaluate the role of CsA on Ca(2+) signaling pathway in intact cells of the porcine proximal tubule-like cell line LLC-PK1; the assay of the in vitro activity of the plasma membrane Ca(2+) pump (PMCA) was carried out through the preparation and isolation of membranes. The addition of CsA to incubation medium at doses ranging from 0.1 to 2 microM did not change the basal level of intracellular calcium ([Ca(2+)](i)), whereas it affected the [Ca(2+)](i) response to thapsigargin (TG), a powerful inhibitor of microsomal Ca(2+) pump. In control studies, 5 microM TG produced a biphasic response: [Ca(2+)](i) peaked with a 60-s lag, and it then declined to a plateau of elevated [Ca(2+)](i), which remains above basal. However, it became evident that CsA strengthened the Ca(2+) response to TG because the addition of 5 microM TG to cells exposed to 400 nM CsA did not affect the peak response to TG, but it markedly affected the subsequent sustained phase ([Ca(2+)](i) = 156 +/- 4.84 versus 130 +/- 3.28 nmol, mean +/- SEM, n = 6, P < 0.001). In membrane preparations, 200 nM CsA brought about, in the presence of 10 microM calmodulin (CaM), a significant decrease of plasma membrane Ca(2+) pump (PMCA) activity (46.96 +/- 0.26 versus 53.48 +/- 1.96 nmol x mg of protein(-1) x min(-1), n = 6, P < 0.02), a value similar to that obtained in the presence of equimolar amounts of cyclosporine H (CsH), a non-immunosuppressive analogue of CsA. These findings suggest that in this cell line CsA affects the Ca(2+) export pathway through the reduction of the PMCA activity with consequent amplification and strengthening of [Ca(2+)](i) response after exposure to agents that trigger intracellular Ca(2+) release. The increased cell sensitivity during Ca(2+) signaling events ensuing from the impairment of this "defense system" may be regarded as one of the basic mechanisms involved in the development of the side effects induced by CsA. PMID- 12761242 TI - The PI3-kinase-Akt pathway promotes mesangial cell survival and inhibits apoptosis in vitro via NF-kappa B and Bad. AB - While the serine/threonine protein kinase Akt has attracted attention as a mediator of survival (anti-apoptotic) signal, the regulation and function of the PI3-kinase-Akt pathway in mesangial cells is not well known. To explore the significance of the PI3-kinase-Akt pathway, this study used PI3-kinase inhibitors (Wortmannin and LY294002) and recombinant adenoviruses encoding a dominant-active mutant of Akt (AxCAmyrAkt) and a dominant-negative mutant of Akt (AxCAAkt-AA) in cultured rat mesangial cells. Apoptotic signals were measured by nucleosomal laddering of DNA, caspase 3 assay, and cell death detection ELISA. The PI3 kinase inhibitors and dominant-negative mutant of Akt increased the apoptotic signals in the presence of platelet-derived growth factor (PDGF), while the dominant-active mutant of Akt prevented apoptosis induced by a serum-free medium. In separate experiments, we further investigated downstream signals of Akt in mesangial cells. While PDGF activated NF-kappa B and phosphorylated Bad, these reactions were inhibited by overexpression of the dominant-negative mutant of Akt as well as the PI3-kinase inhibitors. These data indicate, firstly, that Akt is phosphorylated by PDGF, and secondly, that the activated Akt prevents apoptotic changes via activation of NF-kappa B and phosphorylation of Bad in mesangial cells. This study investigated whether it is Bad phosphorylation or NF-kappa B activation that provides the anti-apoptotic effects of Akt, and the data suggested that NF-kappa B is probably the principal contributor to the downstream activation of the PI3-kinase-Akt pathway. The findings suggest that the PI3 kinase-Akt pathway acts as a survival signal and plays a key role in the regulation of apoptotic change in mesangial cells principally via NF-kappa B. PMID- 12761244 TI - AngRem104, an angiotensin II-induced novel upregulated gene in human mesangial cells, is potentially involved in the regulation of fibronectin expression. AB - Accumulation of extracellular matrix (ECM) in the glomerular mesangium is a common feature of many progressive renal diseases. Angiotensin II (AngII) plays important roles in the proliferation of glomerular mesangial cells (MC) as well as the synthesis of ECM such as fibronectin (FN) and collagens. However, the precise molecular signals responsible for these effects are unknown. To explore possible molecule mechanism of ECM accumulation related to AngII, suppression subtractive hybridization (SSH) was performed to screen and identify upregulated genes induced by AngII in cultured human MC. A novel gene, AngRem104 (GenBank accession number, AF367870), was isolated. The full-length cDNA of AngRem104 is 1690 bp, and it contains a 1041-bp open reading frame (ORF) encoding 347 amino acid residues with a predicted molecular mass of 37.2 kD. AngRem104 widely expressed in human heart, placenta, liver, muscle, kidney, and pancreas. Moreover, AngRem104 was found in human glomeruli and tubule by in situ hybridization. In human MC, the upregulation of AngRem104 induced by AngII was time-dependent, and it was dose-dependently blocked by AngII type 1 receptor antagonist (AT1RA), Losartan. The subcellular localization detected by AngRem104 pEGFP fusion protein revealed that AngRem104 was a nuclear protein. Interestingly, when AngRem104 was overexpressed by transfection of its sense construct, cDNA Microarray showed that two of the ECM-related genes, i.e., human mRNA for FN and integrin-beta-1 (FN receptor), dramatically upregulated their expressions. Furthermore, AngRem104 could regulate the expression of FN induced by AngII, which were detected by RT-PCR and quantitative real-time PCR, when AngRem104 was overexpressed. It is concluded that AngRem104 is a novel human gene potentially involved in the regulation of FN induced by AngII in human MC. These findings may provide new insights into mechanisms of glomerular sclerosis associated with AngII. PMID- 12761245 TI - Cell biological and biochemical characterization of drebrin complexes in mesangial cells and podocytes of renal glomeruli. AB - Drebrins are actin-binding proteins (ABP) initially identified in and thought to be specific for neuronal cells, where they appear to contribute to the formation of cell processes. Recent studies have also detected the isoform drebrin E2 in a wide range of non-neuronal cell types, notably in and near actin-rich lamellipodia and filopodia. The present study demonstrates drebrin enrichment in renal glomeruli. Immunohistochemistry and double-label confocal laser scanning microscopy have shown intense drebrin reactions in the mesangial cells of diverse mammalian species. In adult human and bovine kidneys, drebrin is, in addition, markedly enriched in the foot processes of podocytes, as also demonstrable by immunoelectron microscopy. By contrast, the podocytes of rodent glomeruli appear to contain significant drebrin concentrations only during early developmental stages. In differentiated murine podocytes cultured in vitro, however, drebrin is concentrated in the cell processes, where it partially codistributes with actin and other ABP. In biochemical analyses using protein extracts from renal cortices, large (approximately 20S) complexes ("drebrosomes") were found containing drebrin and actin. These findings confirm and extend our hypothesis that drebrin is involved in the regulation of actin dynamics also outside the nervous system. Clearly, drebrin has to be added to the ensemble of ABP regulating the actomyosin system and the dynamics of mesangial cells and foot processes in podocytes. PMID- 12761246 TI - Lead-induced downregulation of soluble guanylate cyclase in isolated rat aortic segments mediated by reactive oxygen species and cyclooxygenase-2. AB - Lead exposure is a known cause of hypertension. Although most studies have focused on lead-induced endothelial dysfunction and on the involvement of reactive oxygen species (ROS), it has been recently demonstrated that the vascular wall of lead-exposed rats has both an altered the endothelium independent relaxing response and a reduced expression of soluble guanylate cyclase (sGC). The aim of the present study was to determine in in vitro incubated rat isolated aortic segments if lead downregulates sGC expression, analyzing the involvement of ROS and cyclooxygenase-2 (COX-2). The experiments were performed in isolated aortic segments from Wistar rats that were incubated with lead for 24 h. Lead significantly reduced sGC-beta(1) subunit expression in a concentration-dependent manner. The maximal reduction in sGC-beta(1) subunit expression was achieved with 1 ppm lead. Vitamin C (30 micromol/L) partially restored sGC-beta( 1) subunit expression in lead (1 ppm)-exposed aortic segments. A similar protection of sGC-beta(1) subunit expression was obtained with both a protein kinase A inhibitor, H89 (1 micromol/L) and with rofecoxib (1 micromol/L), an inhibitor of COX-2 activity. Moreover, lead exposure increased COX-2 expression in the arterial wall. While vitamin C reduced both COX-2 expression and superoxide anion production related to lead exposure, rofecoxib failed to modify superoxide anion generation in lead-incubated aortic segments. In conclusion, the present results suggest the involvement of ROS and COX-2 in the downexpression of sGC-beta(1) subunit induced by lead in the rat vascular wall. PMID- 12761247 TI - Oxidized LDL and its compound lysophosphatidylcholine potentiate AngII-induced vasoconstriction by stimulation of RhoA. AB - RhoA stimulates vascular tone by increasing smooth muscle Ca(2+) sensitivity, e.g., in atherosclerosis. This study was an investigation of the influence of oxidized LDL (OxLDL), which accumulates in atherosclerotic plaques, on vascular tone induced by angiotensin II (AngII), with particular emphasis on the RhoA pathway. OxLDL had no influence on unstimulated vascular tone of isolated rabbit aorta, but it potentiated contractile responses induced by AngII. The Ca(2+) antagonist felodipin partially prevented potentiation of contractile responses, whereas the AT(1) receptor antagonist losartan blunted AngII responses in presence and in absence of OxLDL. Rho-kinase inhibition by Y27632 abolished potentiation of contractile responses, and RhoA inhibition by C3-like transferase partially prevented it, suggesting that OxLDL activated RhoA. Activation of RhoA was further analyzed by detection of its translocation to the cell membrane after stimulation with OxLDL. Western blot analysis of aorta homogenates, as well as direct visualization in cultured smooth muscle cells using confocal laser scan microscopy, revealed that OxLDL potently activated RhoA. The effect of OxLDL was mimicked by its compound lysophosphatidylcholine, and C3 inhibited both lysophosphatidylcholine and OxLDL-induced RhoA stimulation. In conclusion, OxLDL stimulates the RhoA pathway, resulting in potentiation of AngII-induced vasoconstriction. Lysophosphatidylcholine mimics the OxLDL effect, consistent with a causal role of this OxLDL compound. Stimulation of RhoA by OxLDL may contribute to vasospasm in atherosclerotic arteries. PMID- 12761248 TI - The effects of weight loss on renal function in patients with severe obesity. AB - Severe obesity is associated with increased renal plasma flow (RPF) and glomerular filtration rate (GFR). The aim of the present study was to examine whether weight loss may reverse glomerular dysfunction in obese subjects without overt renal disease. Renal glomerular function was studied in eight subjects with severe obesity (body mass index [BMI] 48.0 +/- 2.4) before and after weight loss. Nine healthy subjects served as controls. GFR and RPF were determined by measuring inulin and PAH clearance. In the obese group, GFR (145 +/- 14 ml/min) and RPF (803 +/- 39 ml/min) exceeded the control value by 61% (90 +/- 5 ml/min, P = 0.001) and 32% (610 +/- 41 ml/min, P < 0.005), respectively. Consequently, filtration fraction was increased. Mean arterial pressure, although normal, was higher than in the control group (101 +/- 4 versus 86 +/- 2 mmHg, P < 0.01). After weight loss, BMI decreased by 32 +/- 4%, to 32.1 +/- 1.5 (P = 0.001). GFR and RPF decreased to 110 +/- 7 ml/min (P = 0.01) and 698 +/- 42 ml/min (P < 0.02), respectively. Albumin excretion rate decreased from 16 microg/min (range, 4 to 152 microg/min) to 5 microg/min (range, 3 to 37 microg/min) (P < 0.01). Fractional clearance of albumin decreased from 3.2 x 10(-6) (range, 1.1 to 23 x 10(-6)) to 1.2 x 10(-6) (range, 0.5 to 6.8 x 10(-6)) (P < 0.02). This study shows that obesity-related glomerular hyperfiltration ameliorates after weight loss. The improvement in hyperfiltration may prevent the development of overt obesity related glomerulopathy. PMID- 12761249 TI - Plasminogen activator inhibitor-1 is a significant determinant of renal injury in experimental crescentic glomerulonephritis. AB - Crescentic glomerulonephritis is characterized by glomerular fibrin deposition, and experimental crescentic glomerulonephritis has been shown to be fibrin dependent. Net fibrin deposition is a balance between activation of the coagulation system causing glomerular fibrin deposition and fibrin removal by the plasminogen-plasmin (fibrinolytic) system. Plasminogen activator inhibitor-1 (PAI 1) inhibits fibrinolysis by inhibiting plasminogen activators and has effects on leukocyte recruitment and matrix deposition. To test the hypothesis that the presence of PAI-1 and its levels were a determinant of injury in crescentic glomerulonephritis, accelerated anti-glomerular basement membrane glomerulonephritis was induced in mice genetically deficient in PAI-1 (PAI-1 -/ ), PAI-1 heterozygotes (PAI-1 +/-), and mice engineered to overexpress PAI-1 (PAI 1 tg). Compared with strain-matched genetically normal animals, PAI-1 -/- mice with glomerulonephritis developed fewer glomerular crescents, less glomerular fibrin deposition, fewer infiltrating leukocytes, and less renal collagen accumulation at day 14 of disease. The reduction in disease persisted at day 28, when injury had become more established. In contrast, mice overexpressing the PAI 1 gene (PAI-1 tg), that have basal plasma and renal PAI-1 levels several times, normal developed increased glomerular crescent formation, more glomerular fibrin deposition, increased numbers of infiltrating leukocytes, and more renal collagen at both time points. These studies demonstrate that PAI-1 is a determinant of glomerular fibrin deposition and renal injury in crescentic glomerulonephritis. PMID- 12761250 TI - Anti-monocyte chemoattractant protein-1 gene therapy attenuates renal injury induced by protein-overload proteinuria. AB - It has been postulated that protein filtered through glomeruli activates tubular epithelial cells, which secrete vasoactive and inflammatory substances including chemokines, leading to tubulointerstitial renal injury. The present study was designed to investigate the role of monocyte chemoattractant protein-1 (MCP-1) in this process and to evaluate the effectiveness of a kidney-targeted gene transfer technique using hydrodynamic pressure. Naked plasmid encoding 7ND (an MCP-1 antagonist) or a control plasmid was introduced into the left kidney of rats. Three days after gene transfer (day 0), intraperitoneal administration of bovine serum albumin (10 mg/g body wt per day) was started and continued for 14 or 21 d. RT-PCR showed that 7ND mRNA was expressed only in the gene-transfected kidney. Immunostaining showed that 7ND protein was localized in the interstitial cells. Macrophage infiltration was significantly reduced in the left kidney of rats treated with 7ND on days 14 and 21. In the right kidney, such effects were not observed. 7ND also attenuated tubular damage and decreased the number of apoptotic cells. Computer-assisted analysis revealed that the areas positively stained for alpha-smooth muscle actin (alpha SMA), fibronectin-EDA, type I collagen, and collagen fibrils were significantly reduced in the 7ND-treated kidney on day 21. Furthermore, 7ND gene therapy significantly reduced MCP-1 and TGF-beta 1 mRNA expression. These results demonstrate that MCP-1 plays an important role in the development of tubulointerstitial inflammation, tubular damage, and fibrosis induced by proteinuria. The fact that 7ND gene therapy had little effect on the contralateral kidney indicates that 7ND acted locally. This strategy may have a potential usefulness as a gene therapy against tubulointerstitial renal injury. PMID- 12761251 TI - Nephrogenesis is induced by partial nephrectomy in the elasmobranch Leucoraja erinacea. AB - The mammalian kidney responds to partial nephrectomy with glomerular and tubular hypertrophy, but without renal regeneration. In contrast, renal regeneration in lower vertebrates is known to occur. Understanding the underlying mechanisms of renal regeneration is highly important; however, a serviceable animal model has not been developed. A neonephrogenic zone has been identified in the European lesser spotted dogfish, Scyliorhinus caniculus (Hentschel H. Am J Anat 190: 309 333, 1991), as well as in the spiny dogfish Squalus acanthias and the little skate, Leucoraja erinacea. The zone features the production of new nephrons complete with a countercurrent system. To analyze this nephrogenic region of elasmobranch fish further, a renal reduction model was established. The neonephrogenic zone in the adult kidney of the little skate resembles the embryonic metanephric kidney and contains stem cell-like mesenchymal cells, tips of the branching collecting duct system, and outgrowth of the arterial system. Four stages of nephron development were analyzed by serial sections and defined: stage I, aggregated mesenchymal cells; stage II, S-shaped body-like structure with high-prismatic epithelial cells; stage III, segmental nephron segregation; stage IV, functioning nephron. The stages were analyzed after partial nephrectomy. In addition, cell proliferation was assessed by incorporation of bromo-deoxyuridine (BrdU). New nephrons developed in animals undergoing partial nephrectomy. Growth was greatly stimulated in the nephrogenic zone, both in the remnant tissue and in the contralateral kidney within 10 wk. Mesenchymal cell aggregates increased significantly per renal cross-section compared with controls (stage I, 0.64 +/- 0.28 versus 0.27 +/- 0.25; P < 0.005; n = 10 animals per group). The same was the case for S-shaped body-like cysts (stage II, 0.24 +/- 0.19 versus 0.08 +/- 0.09; P < 0.02). Cellular proliferation in the neonephrogenic zone of the contralateral kidney was also greatly enhanced (14.42 +/- 3.26 versus 2.64 +/- 1.08 BrdU-positive cells per cross-section, P < 0.001). It is concluded that the skate possesses a nephrogenic zone containing stem cell like mesenchymal cells during its entire life. Partial nephrectomy induces renal growth by accelerating nephrogenesis. This unique model may facilitate understanding renal regeneration. PMID- 12761252 TI - A gene locus for steroid-resistant nephrotic syndrome with deafness maps to chromosome 14q24.2. AB - Steroid-resistant nephrotic syndrome (SRNS) leads to end-stage renal disease (ESRD) in childhood or young adulthood. Positional cloning for genes causing SRNS has opened the first insights into the understanding of its pathogenesis. This study reports a genome-wide search for linkage in a consanguineous Palestinian kindred with SRNS and deafness and detection of a region of homozygosity on chromosome 14q24.2. Multipoint analysis of 12 markers used for further fine mapping resulted in a LOD score Z(max) of 4.12 (theta = 0) for marker D14S1025 and a two-point LOD score of Z(max) = 3.46 (theta = 0) for marker D14S77. Lack of homozygosity defined D14S1065 and D14S273 as flanking markers to a 10.7 cM interval. The identification of the responsible gene will provide new insights into the molecular basis of nephrotic syndrome and sensorineural deafness. PMID- 12761253 TI - Activin a produced by ureteric bud is a differentiation factor for metanephric mesenchyme. AB - The present study was conducted to investigate the role of the activin follistatin system in the development of metanephros. Organ culture system and cultured metanephric mesenchymal cells were used to address this issue. Activin A was localized in ureteric bud. Activin type II receptor was localized in ureteric bud as well as metanephric mesenchyme. In an organ culture system, exogenous activin A reduced the size of cultured metanephroi, delayed ureteric bud branching, and enlarged the tips of ureteric bud. Follistatin, an antagonist of activin A was used to clarify the role of endogenous activin A. Exogenous follistatin enlarged the size of cultured metanephroi, increased ureteric bud branching, and promoted cell growth in ureteric bud. Blockade of activin signaling by adenoviral transfection of dominantly negative activin mutant receptor mimics the effect of follistatin. In cultured metanephric mesenchymal cells, activin A promoted cell growth; conversely, follistatin induced apoptosis. Furthermore, activin A induced the expressions of epithelial differentiation markers in these cells. These results suggest that activin A produced by ureteric bud is not only an important regulator of ureteric bud branching, but also a differentiation factor for metanephric mesenchyme during kidney development. PMID- 12761254 TI - Inhibition of renal fibrosis by gene transfer of inducible Smad7 using ultrasound microbubble system in rat UUO model. AB - TGF-beta is a key mediator in renal fibrosis. Kidney-targeted gene therapy with anti-TGF-beta strategies is expected to have therapeutic potential, but this has been hampered by concerns over the safety and practicability of viral vectors and the inefficiency of nonviral transfection techniques. The present study explored the potential role of TGF-beta/Smad signaling in renal fibrosis in vivo and developed a safe and effective gene therapy to specifically block TGF-beta signaling and renal fibrosis in a rat unilateral ureteral obstruction (UUO) model by transferring a doxycycline-regulated Smad7 gene or control empty vectors using an ultrasound-microbubble (Optison)-mediated system. The Smad7 transgene expression was tightly controlled by addition of doxycycline in the daily drinking water. Groups of six rats were sacrificed at day 7, and the transfection rate, Smad7 transgene expression, and tubulointerstitial fibrosis including alpha smooth muscle actin and collagen matrix mRNA and protein expression were determined. Compared with the non-ultrasound treatment, the combination of ultrasound with Optison largely increased the transfection rate of FITC-ODN and Smad7 transgene expression up to a 1000-fold, and this was found in all kidney tissues. Compared with normal rats, Smad7 expression within the UUO kidney was significantly reduced, and this was associated with up to a sixfold increase in Smad2 and Smad3 activation and severe tubulointerstitial fibrosis. In contrast, treatment with inducible Smad7 resulted in a fivefold increase in Smad7 expression with complete inhibition of Smad2 and Smad3 activation and tubulointerstitial fibrosis in terms of tubulointerstitial myofibroblast accumulation (85% downward arrow ) and collagen I and III mRNA and protein expression (60 to 70% downward arrow ). In conclusion, the ultrasound-mediated inducible Smad7 gene transfer is a safe, effective, and controllable gene therapy. TGF-beta-mediated renal fibrosis is regulated positively by Smad2/3, but negatively by Smad7. Target blockade of TGF-beta/Smad signaling by expression of Smad7 may provide a new therapeutic potential for renal fibrosis. PMID- 12761255 TI - Distant effects of experimental renal ischemia/reperfusion injury. AB - Acute renal failure results in significant morbidity and mortality, yet renal failure is not the usual cause of death in the clinical situation. We have previously reported systemic increases in the inflammatory mediators tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 (IL-1) after renal ischemia in the mouse. In the present study, an animal model of bilateral renal ischemia was used to test the hypothesis that cytokines released with renal ischemia have effects on other organ systems. Increased levels of immunoreactive TNF-alpha and IL-1 and intercellular adhesion molecule-1 mRNA were found in the heart after renal ischemia in the rat. This was accompanied by increases in myeloperoxidase activity, an index of tissue leukocyte infiltration, in the heart as well as the liver and lung. Functional changes in the heart 48 h after renal ischemia included increases in left ventricular end diastolic diameter, left ventricular end systolic diameter, and decreased fractional shortening by echocardiography. Evidence of apoptosis of cardiac cells was also found 48 h after an abbreviated period of renal ischemia insufficient to induce azotemia but not bilateral nephrectomy (which resulted in significant renal failure), suggesting that renal ischemia but not uremia is necessary for the apoptosis observed. It was also found that blocking the action of TNF-alpha limited cardiac apoptosis. Renal ischemia results in distant effects and the alterations observed in the heart may be important in the morbidity and mortality observed clinically. PMID- 12761256 TI - BMP-7 is an efficacious treatment of vascular calcification in a murine model of atherosclerosis and chronic renal failure. AB - Chronic renal failure is complicated by high cardiovascular mortality. One key contributor to this mortality is vascular calcification, for which no therapy currently exists. Bone morphogenetic protein 7 is an essential renal morphogen that maintains renal tubular differentiation in the adult and is downregulated in renal failure. Several studies have demonstrated its efficacy in treating various renal diseases in rodents, and it was hypothesized that it would also be an effective treatment of vascular calcification in this setting. Uremia was imposed on LDL receptor null mice (a model of atherosclerosis), which were then treated with bone morphogenetic protein 7 for 15 wk. Uremic animals had increased vascular calcification by histology and chemical analysis. Calcification in treated animals was similar to or less than non-uremic control animals. Cells exhibiting an osteoblast-like phenotype in the vessel wall may be important in the etiology of vascular calcification. Expression of osteocalcin was assessed as a marker of osteoblastic function, and it is shown that it is increased in untreated uremic animals but downregulated to levels similar to non-uremic control animals with treatment. The data are compatible with bone morphogenetic protein 7 deficiency as a pathophysiologic factor in chronic renal failure, and they demonstrate its efficacy as a potential treatment of vascular calcification. PMID- 12761257 TI - The contribution of increased diabetes prevalence and improved myocardial infarction and stroke survival to the increase in treated end-stage renal disease. AB - This study examined the extent to which the greater than threefold increase in number of treated end-stage renal disease (ESRD) cases between 1978 and 1991 is explained by increases in the prevalence of diabetes, myocardial infarction (MI) and stroke survivors, and US population size. The change in number of persons in the United States with diabetes, a history of MI or stroke, and without these conditions was estimated for 1978 and 1991 using the Second and Third National Health and Nutrition Examination Surveys. The treated ESRD incidence rate and the increase in ESRD treatment attributable to each of these populations were calculated using these estimates and data from the United States Renal Data System. In the United States, there were an estimated 4.3 and 1.2 million more persons with diabetes and a history of MI or stroke, respectively, 22.7 million more persons without these conditions, and 36,881 more incident treated ESRD cases in 1991 compared with 1978. In 1991, treated ESRD incidence rates among persons with diabetes, a history of MI or stroke, and without these conditions were 2567, 1463, and 153 cases per million person-years, respectively. In 1991, 10,183 cases of the additional 36,881 treated ESRD cases (27.6%) resulted from the higher prevalence of diabetes; 1775 (4.8%) from increased MI and stroke survival; and 2904 (7.9%) resulted from growth of the US population without these conditions. The increasing number of treated ESRD cases in the United States is partly explained by the increase in diabetes prevalence and US population growth but only minimally by MI and stroke survival. PMID- 12761258 TI - Treatment of IgA nephropathy with ACE inhibitors: a randomized and controlled trial. AB - Some retrospective studies have suggested a beneficial influence of angiotensin converting enzyme (ACE) inhibitors on the progression of IgA nephropathy (IgAN), but prospective and controlled studies demonstrating this effect are lacking. Forty-four patients with biopsy-proven IgAN, proteinuria > or = 0.5 g/d, and serum creatinine (SCr) < or = 1.5 mg/dl were randomly assigned either to receive enalapril (n = 23) or to a control group (n = 21) in whom BP was controlled with antihypertensives other than ACE inhibitors. Primary outcome was renal survival estimated by a 50% increase in baseline SCr. Secondary outcomes were the presence of a SCr > 1.5 mg/dl at the last visit and the evolution of proteinuria. Baseline clinical findings were similar at baseline between enalapril-treated and control group, and there were no differences in BP control during follow-up. Mean follow up was 78 +/- 37 mo in the enalapril group and 74 +/- 36 mo in the control group. Three patients (13%) in the enalapril group and 12 (57%) in the control group reached the primary end point (P < 0.05). Kaplan-Meier renal survival was significantly better in enalapril group than in control group: 100% versus 70% after 4 yr and 92% versus 55% after 7 yr (P < 0.05). Three patients in the enalapril group (13%) and 11 (52%) in the control group showed SCr > 1.5 mg/dl at the last visit (P < 0.05). Proteinuria significantly decreased in the enalapril group, whereas it tended to increase in the control group (P < 0.001 between groups). In conclusion, ACE inhibitors significantly improve renal survival in proteinuric IgAN with normal or moderately reduced renal function. PMID- 12761259 TI - Predictors of renal and patient outcomes in atheroembolic renal disease: a prospective study. AB - Atheroembolic renal disease (AERD) is part of a multisystemic disease accompanied by high cardiovascular comorbidity and mortality. Interrelationships between traditional risk factors for atherosclerosis, vascular comorbidities, precipitating factors, and markers of clinical severity of the disease in determining outcome remain poorly understood. Patients with AERD presenting to a single center between 1996 and 2002 were followed-up with prospective collection of clinical and biochemical data. The major outcomes included end-stage renal disease (ESRD) and death. Ninety-five patients were identified (81 male). AERD was iatrogenic in 87%. Mean age was 71.4 yr. Twenty-three patients (24%) developed ESRD; 36 patients (37.9%) died. Cox regression analysis showed that significant independent predictors of ESRD were long-standing hypertension (hazard ratio [HR] = 1.1; P < 0.001) and preexisting chronic renal impairment (HR = 2.12; P = 0.02); use of statins was independently associated with decreased risk of ESRD (HR = 0.02; P = 0.003). Age (HR = 1.09; P = 0.009), diabetes (HR = 2.55; P = 0.034), and ESRD (HR = 2.21; P = 0.029) were independent risk factors for patient mortality; male gender was independently associated with decreased risk of death (HR = 0.27; P = 0.007). Cardiovascular comorbidities, precipitating factors, and clinical severity of AERD had no prognostic impact on renal and patient survival. It is concluded that AERD has a strong clinical impact on patient and renal survival. The study clearly shows the importance of preexisting chronic renal impairment in determining both renal and patient outcome, this latter being mediated by the development of ESRD. The protective effect of statins on the development of ESRD should be evaluated in a prospective study. PMID- 12761260 TI - Ankle-brachial blood pressure index predicts all-cause and cardiovascular mortality in hemodialysis patients. AB - A reduction in ankle-brachial BP index (ABPI) is associated with generalized atherosclerotic diseases and predicts cardiovascular mortality and morbidity in several patient populations. However, a large-scale analysis of ABPI is lacking for hemodialysis (HD) patients, and its use in this population is not fully validated. A cohort of 1010 Japanese patients undergoing chronic hemodialysis was studied between November 1999 and May 2002. Mean age at entry was 60.6 +/- 12.5 yr, and duration of follow-up was 22.3 +/- 5.6 mo. Patients were stratified into five groups (< 0.9, > or = 0.9 to < 1.0, > or = 1.0 to < 1.1, > or = 1.1 to < 1.3, and > or = 1.3) by ABPI measured at entry by an oscillometric method. The frequency distribution of ABPI was 16.5% of patients < 0.9, 8.6% of patients > or = 0.9 to < 1.0, 16.9% of patients 1.0 > or = to < 1.1, and 47.0% of patients > or 1.1 to < 1.3, whereas 10.9% of patients had an abnormally high ABPI (> or = 1.3). The relative risk of a history of diabetes mellitus (DM), cardiovascular, and cerebrovascular disease was significantly higher in patients with lower ABPI than those with ABPI > or = 1.1 to <1.3. During the study period, 77 cardiovascular and 41 noncardiovascular fatal events occurred. On the basis of Cox proportional hazards regression analysis, ABPI emerged as a strong independent predictor of all-cause and cardiovascular mortality. After adjustment for confounding variables, the hazard ratio (HR) for ABPI < 0.9 was 4.04 (95% confidence interval, 2.38 to 6.95) for all-cause mortality and 5.90 (2.83 to 12.29) for cardiovascular mortality. Even those with modest reductions in the ABPI (> or = 0.9 to <1.1) appeared to be at increased risk. Patients having abnormally high ABPI (> or = 1.3) also had poor prognosis (HR, 2.33 [1.11 to 4.89] and 3.04 [1.14 to 8.12] for all-cause and cardiovascular mortality, respectively). Thus, the present findings validate ABPI as a powerful and independent predictor for all cause and cardiovascular mortality among hemodialysis patients. PMID- 12761261 TI - Blood flow limitation in vivo of small solute transfer during peritoneal dialysis in rats. AB - The aim of this study was to determine whether or to what extent transperitoneal flux of small solutes is reduced at low blood flows during peritoneal dialysis (PD) in rats. Peritoneal blood flow reductions were achieved by bleeding anesthetized (300 g) rats by 25% of their blood volume. After bleeding, a 2 h PD dwell was started using standard PD fluid. The permeability-surface area product (PS) for (51)Cr-EDTA and glucose were assessed, as well as the transperitoneal clearance (Cl) of albumin. Control animals were not bled. After bleeding, peritoneal blood flow declined from 145 +/- 17 perfusion units (PU) to 59 +/- 12 PU (P = 0.001). Concomitant with this reduction, PS for (51)Cr-EDTA fell from 0.284 +/- 0.01 ml/min to 0.216 +/- 0.01 ml/min (P = 0.006) and PS for glucose from 0.338 +/- 0.02 ml/min to 0.294 +/- 0.01 ml/min (P = 0.046). Mean arterial BP (MAP) dropped from 133 +/- 4 mmHg to 61 +/- 5 mmHg (P = 0.008). Cl of albumin fell largely in proportion to the estimated capillary hydrostatic pressure drop, i.e., from 6.1 +/- 0.7 microl/min to 2.3 +/- 0.3 microl/min (P = 0.001). The results demonstrate that the transperitoneal clearances of small solutes are blood flow limited during PD, when peritoneal perfusion is markedly reduced. The level of flow limitation was, however, much lower than expected and observed in other tissues. Albumin transport, which is not blood flow limited, was reduced largely in proportion to the calculated capillary hydrostatic pressure decrease. PMID- 12761262 TI - Effect of pravastatin on loss of renal function in people with moderate chronic renal insufficiency and cardiovascular disease. AB - Limited data suggest that HMG-CoA reductase inhibitors (statins) may slow loss of renal function in individuals with chronic renal insufficiency. This study was conducted to determine whether pravastatin reduced rates of loss of renal function in people with moderate chronic renal insufficiency. This was a post hoc subgroup analysis of a randomized double-blind placebo controlled trial. Data were analyzed from the CARE study (a randomized trial of pravastatin versus placebo in 4159 participants with previous myocardial infarction and total plasma cholesterol < 240 mg/dl). Participants with estimated GFR (MDRD-GFR) < 60 ml/min per 1.73 m(2) body surface area at baseline were considered to have moderate chronic renal insufficiency. Multivariate regression was used to calculate rates of decline in MDRD-GFR for individuals receiving pravastatin and placebo, controlling for prospectively determined covariates that might influence rates of renal function loss. Change in renal function could be calculated in 3384 individuals, of whom 690 (20.4%) had MDRD-GFR < 60 ml/min per 1.73 m(2) and were eligible for inclusion. Among all individuals with MDRD-GFR < 60 ml/min per 1.73 m(2)), the MDRD-GFR decline in the pravastatin group was not significantly different from that in the placebo group (0.1 ml/min per 1.73 m(2)/yr slower; 95% CI, -0.2 to 0.4; P = 0.49). However, there was a significant stepwise inverse relation between MDRD-GFR before treatment and slowing of renal function loss with pravastatin use, with more benefit in those with lower MDRD-GFR at baseline (P = 0.04). Rate of change in MDRD-GFR in the pravastatin group was 0.6 ml/min per 1.73 m(2)/yr slower than placebo (95% CI, -0.1 to 1.2; P = 0.07) in those with MDRD-GFR < 50 ml/min, and 2.5 ml/min per 1.73 m(2)/yr slower (95% CI, 1.4 to 3.6 slower; P = 0.0001) in those with MDRD-GFR < 40 ml/min per 1.73 m(2)/yr. Pravastatin also reduced rates of renal loss to a greater extent in participants with than without proteinuria at baseline (P = 0.006). It is concluded that pravastatin may slow renal function loss in individuals with moderate to severe kidney disease, especially those with proteinuria. These findings require confirmation by a large randomized trial conducted specifically in people with chronic renal insufficiency. PMID- 12761263 TI - Plasma pentosidine is associated with inflammation and malnutrition in end-stage renal disease patients starting on dialysis therapy. AB - Pentosidine is an advanced glycation end-product (AGE), formed by glycosylation and oxidation, that accumulates markedly in end-stage renal disease (ESRD). It has been speculated that AGE and carbonyl stress contributes to long-term complications such as cardiovascular disease (CVD) in ESRD patients. This study determined plasma levels of pentosidine as well as the presence of inflammation (CRP > or = 10 mg/L), clinical CVD (CVD(clin)), and malnutrition (subjective global assessment [SGA] > 1) in a cohort of 191 ESRD patients, median age of 55 yr (range, 23 to 70 yr) and median GFR = 7 ml/min (range, 2 to 17 ml/min), close to start of renal replacement therapy. Fifty-one elderly subjects, median age of 82 yr (range, 71 to 110 yr), with mild renal impairment, median GFR = 67 ml/min (range, 38 to 113 ml/min), were also studied for comparative analysis of plasma pentosidine. The plasma pentosidine content was elevated in all patients compared with the levels in the elderly subjects and were negatively correlated with GFR both in the ESRD patients (Rho = -0.24; P < 0.01; n = 159) and in the elderly subjects (Rho = -0.31; P < 0.05). Moreover, the plasma pentosidine content was correlated with age in the ESRD patients (Rho = 0.26; P < 0.001) and in the elderly subjects (Rho = 0.44; P < 0.001). The 63 malnourished ESRD patients (35%) had a significantly higher (P < 0.05) median plasma pentosidine than the well nourished patients (39 versus 27 pmol/mg albumin). Similarly, 73 inflamed patients (38%) had a significantly higher (P < 0.001) median pentosidine content compared with 118 non-inflamed patients (37 versus 24 pmol/mg albumin). Also, the plasma pentosidine content showed weak but significant positive correlations with CRP (Rho = 0.28; P < 0.0001), fibrinogen (Rho = 0.23; P < 0.01; n = 126), IL-6 (Rho = 0.22; P < 0.01; n = 169), and soluble vascular cellular adhesion molecule 1 (Rho = 0.38; P < 0.001; n = 74). On the other hand, no significant differences in plasma pentosidine content were noted between the patients with and those without CVD(clin) (32 versus 27 pmol/mg albumin, respectively). Analyses of all cause mortality, by Kaplan-Meier, showed that mortality was not linked to the plasma pentosidine content. Moreover, survival analysis by the Cox regression model showed that age (P < 0.001), diabetes mellitus (P < 0.01), malnutrition (P < 0.01), and CVD(clin) (P < 0.01) independently predicted poor outcome, whereas an elevated plasma pentosidine content did not. The present study shows that an elevated plasma pentosidine content in ESRD patients is significantly associated with both inflammation and malnutrition and confirms that low residual renal function and high age further contribute to an increased plasma pentosidine content. However, in this small cohort, the plasma pentosidine content did not predict outcome. Thus, accumulation of plasma pentosidine is unlikely to be an appropriate clinically useful marker to predict mortality in ESRD patients. PMID- 12761264 TI - A prospective controlled trial on effect of percutaneous transluminal angioplasty on functioning arteriovenous fistulae survival. AB - Balloon angioplasty (PTA) is an established treatment modality for stenosis in dysfunctional arteriovenous fistulae (AVF), although most studies showing efficacy have been retrospective, uncontrolled, and nonrandomized. In addition, it is unknown whether correction of stenosis not associated with significant hemodynamic, functional, and clinical abnormality may improve survival in AVF. This study was a prospective controlled open trial to evaluate whether prophylactic PTA of stenosis not associated with access dysfunction improves survival in native, virgin, radiocephalic forearm AVF. Sixty-two stenotic, functioning AVF, i.e., able to provide adequate dialysis, were enrolled in the study: 30 were allocated to control and 32 to PTA. End points of the study were either AVF thrombosis or surgical revision due to reduction in delivered dialysis dose. Kaplan-Meier analysis showed that PTA improved AVF functional failure-free survival rates (P = 0.012) with a fourfold increase in median survival and a 2.87 fold decrease in risk of failure. Cox proportional hazard model identified PTA as the only variable associated with outcome (P = 0.012). PTA induced an increase in access blood flow rate (Qa) by 323 (236 to 445) ml/min (P < 0.001), suggesting that improved AVF survival is the result of increased Qa. PTA was also associated with a significant decrease in access-related morbidity by approximately halving the risk of hospitalization, central venous catheterization, and thrombectomy (P < 0.05). This study shows that prophylactic PTA of stenosis in functioning forearm AVF improves access survival and decreases access-related morbidity, supporting the usefulness of preventive correction of stenosis before the development of access dysfunction. It also strongly supports surveillance program for early detection of stenosis. PMID- 12761265 TI - Hospitalized psychoses after renal transplantation in the United States: incidence, risk factors, and prognosis. AB - Although it is recommended that renal transplant (RT) candidates routinely undergo screening for mental health-related conditions, national statistics for psychoses after RT have not been reported. This is a historical cohort study of 39,628 renal transplant recipients in the United States Renal Data System between July 1, 1994, and June 30, 1998, and followed until December 31, 1999. Adjusted hazard ratios (AHR) for time to hospitalization for both a primary and secondary discharge diagnosis of psychoses (ICD-9 codes 290.x-299.x) after RT and mortality/graft loss after psychosis were assessed by Cox Regression. In addition, rates of psychosis were compared with 178,986 patients with Medicare as their primary payer who started chronic dialysis from April 1, 1995, to June 29, 1999. The incidence of psychoses was 7.5/1000 person-years (PY) after RT compared with 7.2/1000 PY for all patients on chronic dialysis and 9.6/1000 PY for dialysis patients aged 65 yr or younger. Among RT recipients, graft loss (AHR, 2.97; 95% CI, 2.19 to 4.02), allograft rejection, and cadaveric donation were independently associated with psychosis, which was associated with an increased risk of both death (AHR, 2.09; 95% CI, 1.71 to 2.56; P < 0.001) and graft loss (AHR, 1.79; 95% CI, 1.15 to 2.78; P = 0.01). Graft loss due to noncompliance was significantly more common after psychosis (9.0% versus 3.7% in patients not hospitalized for psychosis; P < 0.001). The incidence of hospitalized psychosis was not substantially higher after RT compared with chronic dialysis patients. Psychoses were independently associated with increased risk of death and graft loss after renal transplantation, possibly mediated through medical non adherence. PMID- 12761266 TI - The change in allograft function among long-term kidney transplant recipients. AB - Long-term kidney allograft survival continues to remain an elusive goal. Kidney transplant recipients are believed to be at high risk for loss of allograft function, and new, potentially non-nephrotoxic immunosuppressive medications are advocated to improve long-term allograft survival. To evaluate the efficacy of such therapeutic interventions, information regarding the change in GFR among kidney transplant recipients with long-term allograft survival is needed. We studied 40,963 transplant recipients between 1987 and 1996 with allograft survival of at least 2 yr in the United States Renal Data System. Linear regression methods were applied to serial GFR estimates after transplantation. The baseline mean GFR at 6 mo after transplantation was 49.6 +/- 15.4 ml/min per 1.73 m(2). During the mean follow-up of 5.7 +/- 2.3 yr, the mean +/- standard error of the change in GFR was -1.66 +/- 6.51 ml/min per 1.73 m(2) per year (median, -0.94 L/min per 1.73 m(2) per year). A total of 12,583 (30%) of patients had improvement in GFR, 8133 (20%) patients had no change in GFR, and 20,247 (50%) patients had decline in GFR. It is concluded that, although most patients had significant impairment of GFR at baseline, the decline in GFR was slow and many patients had either no change or improvement in GFR. Strategies to improve long-term kidney allograft survival that increase baseline allograft function may be more effective than strategies to slow the decline in GFR. PMID- 12761267 TI - Regulatory CD25+ T cells in human kidney transplant recipients. AB - Recent evidence suggests that a population of professional regulatory cells, which limit immune responsiveness, exist in rodents and healthy human subjects. However, their role in disease states remains unclear. A proportion of renal transplant recipients do not demonstrate in vitro reactivity toward their mismatched donor-derived HLA-DR antigens; it was therefore hypothesized that this may be due to such regulatory cells. A cohort of 23 renal transplant recipients was studied at a single institution. In patients with no history of acute rejection, 6 (40%) of 15 demonstrated regulation toward the mismatched HLA-DR allopeptides by CD25(+) cells. By contrast, only one (12.5%) in eight of those with a history of acute rejection demonstrated regulation. Interestingly, if the patient assays were stratified according to initial in vitro immune responsiveness toward the mismatched allopeptides, 8 (47.1%) of 17 of patient assays with low allopeptide responsiveness (alloreactive T cell frequencies less than 60/million) demonstrated regulation of indirect pathway alloresponses by CD25(+) cells, whereas 0 of 8 with higher responses (frequencies greater than 60/million) demonstrated no such regulation (P < 0.05 by chi(2) test). The regulatory cells are present in the circulation as early as 3 mo after transplantation and persist for a number of years, despite conventional immunosuppression. Furthermore, induction treatment with anti-IL-2R mAb did not prevent the development of these regulatory CD25(+) cells. Data from two patients suggest that these cells may also play a role in preventing epitope shifting, implicated in the ongoing immune activation contributing to chronic rejection, and that loss of regulation in a given patient may precede an episode of rejection. PMID- 12761268 TI - CD4+CD25+ regulatory T cells do not significantly contribute to direct pathway hyporesponsiveness in stable renal transplant patients. AB - CD4(+)CD25(+) regulatory T cells have been shown to regulate a variety of autoimmune and allogeneic responses in mice and humans. The role of CD4(+)CD25(+) cells in regulating alloresponses in human transplant recipients remains uncertain. Previous research has demonstrated a reduced frequency of direct pathway donor-specific T cells in renal transplant recipients when compared with the frequency of T cells reactive to an HLA-matched third party. A number of mechanisms have been proposed to account for this finding; the purpose of this study was to determine whether CD4(+)CD25(+) cells play a significant role. Twelve stable renal transplant patients were investigated using limiting dilution assay (LDA) and ELISPOT for interferon-gamma to determine the effect of depleting CD4(+)CD25(+) cells on the direct pathway alloresponse. The percentage of CD4(+)CD25(+) cells in the peripheral blood of the study patients was equivalent to that of healthy controls. Furthermore, in no case did depletion of CD4(+)CD25(+) cells significantly increase the frequency of donor-specific T cells detected by LDA. This was also found with ELISPOT in all except one patient, in whom depletion revealed an increased frequency of alloreactive T cell to both donor and third party. Finally, kinetic analysis of the LDA data did not indicate regulation against donor when compared with third party. It is concluded that the action of CD4(+)CD25(+) regulatory cells is not the main mechanism of donor-specific hyporesponsiveness in the direct pathway of allorecognition. PMID- 12761269 TI - Computerized image analysis of Sirius Red-stained renal allograft biopsies as a surrogate marker to predict long-term allograft function. AB - Chronic allograft nephropathy (CAN) is a major problem in posttransplant management. The lack of a reliable and early surrogate marker of CAN has hampered patient care and research. In this study, the Cortical Fractional Interstitial Fibrosis Volume (V(IntFib)), quantitated with computerized image analysis of Sirius Red-stained protocol biopsies, was examined as a potential surrogate for time to graft failure (TTGF) in 68 renal allograft recipients. At 6 mo posttransplant, V(IntFib) was highly correlated with TTGF (r = 0.64, P < 0.001). Both the Banff Chronic Sum and the Acute Sum Scores were also correlated with TTGF, but less strongly (r = 0.28, P < 0.02; r = 0.35, P < 0.003, respectively). As V(IntFib) was not correlated with the Banff Chronic Score, a multivariate model was created that incorporated V(IntFib) and both Acute and Chronic Banff pathology. This model was highly correlated with TTGF (r = 0.7, P < 0.0001). These findings suggest that V(IntFib) determined by computerized image analysis of Sirius Red-stained protocol biopsies at 6 mo posttransplant, with or without incorporation of Banff acute and chronic scoring, may provide an early surrogate for time to graft failure in renal allograft recipients. PMID- 12761270 TI - The arteriovenous fistula. PMID- 12761271 TI - Homer W. Smith Award lecture. The juxtaglomerular apparatus: from anatomical peculiarity to physiological relevance. PMID- 12761272 TI - Reflections on the past, transitions to the future: the American Society of Nephrology. PMID- 12761273 TI - The skate weighs in on kidney regeneration. PMID- 12761274 TI - Regulatory T cells in kidney transplant recipients: active players but to what extent? PMID- 12761275 TI - Sodium currents in neurons from the rostroventrolateral medulla of the rat. AB - Rapidly inactivating and persistent sodium currents have been characterized in acutely dissociated neurons from the area of rostroventrolateral medulla that included the pre-Botzinger Complex. As demonstrated in many studies in vitro, this area can generate endogenous rhythmic bursting activity. Experiments were performed on neonate and young rats (P1-15). Neurons were investigated using the whole cell voltage-clamp technique. Standard activation and inactivation protocols were used to characterize the steady-state and kinetic properties of the rapidly inactivating sodium current. Slow depolarizing ramp protocols were used to characterize the noninactivating sodium current. The "window" component of the rapidly inactivating sodium current was calculated using mathematical modeling. The persistent sodium current was revealed by subtraction of the window current from the total noninactivating sodium current. Our results provide evidence of the presence of persistent sodium currents in neurons of the rat rostroventrolateral medulla and determine voltage-gated characteristics of activation and inactivation of rapidly inactivating and persistent sodium channels in these neurons. PMID- 12761276 TI - Static and dynamic membrane properties of lateral vestibular nucleus neurons in guinea pig brain stem slices. AB - In vitro intracellular recordings of central vestibular neurons have been restricted so far to the medial vestibular nucleus (MVN). We performed intracellular recordings of large Deiters' neurons in the lateral vestibular nucleus (LVN) to determine their static and dynamic membrane properties, and compare them with those of type A and type B neurons identified in the MVN. Unlike MVN neurons (MVNn), the giant-size LVN neurons (LVNn) form a homogeneous population of cells characterized by sharp spikes, a low-amplitude, biphasic after-hyperpolarization like type B MVNn, but also an A-like rectification like type A MVNn. In accordance with their lower membrane resistance, the sensitivity of LVNn to current injection was lower than that of MVNn over a large range of frequencies. The main difference between LVNn and MVNn was that the Bode plots showing the sensitivity of LVNn as a function of stimulation frequency were flatter than those of MVNn, and displayed a weaker resonance. Furthermore, most LVNn did not show a gradual decrease of their firing rate modulation in the frequency range where it was observed in MVNn. LVNn synchronized their firing with the depolarizing phase of high-frequency sinusoidal current injections. In vivo studies have shown that the MVN would be mainly involved in gaze control, whereas the giant LVNn that project to the spinal cord are involved in the control of posture. We suggest that the difference in the membrane properties of LVNn and MVNn may reflect their specific physiological roles. PMID- 12761278 TI - Neurons in the primate orbitofrontal cortex respond to fat texture independently of viscosity. AB - The primate orbitofrontal cortex (OFC) is a site of convergence from primary taste, olfactory, and somatosensory cortical areas. We describe the responses of a population of single neurons in the OFC that respond to orally applied fat (e.g., safflower oil) and to substances with a similar texture but different chemical composition, such as mineral oil (hydrocarbon) and silicone oil [(Si(CH3)2O)n]. These findings provide evidence that the neurons respond to the oral texture of fat, sensed by the somatosensory system. Use of an oral viscosity stimulus consisting of carboxymethyl-cellulose in the range 1-10,000 centipoise (cP) showed that the responses of these fat-sensitive neurons are not related to stimulus viscosity. Thus a textural component independent of viscosity and related to the slick or oily property is being used to activate these oral fat sensitive neurons. Moreover, a separate population of neurons responds to viscosity (produced, e.g., by the carboxymethyl-cellulose series), but not to fat with the same viscosity. Thus there is a dissociation between texture channels used to sense fat viscosity and non-fat-produced viscosity. Further, free fatty acids such as linoleic acid do not activate these neurons, providing further evidence that the oral fat-sensing mechanism through which these OFC neurons are activated is not gustatory but textural. Most of this population of fat-sensitive neurons receive convergent taste inputs. These results provide evidence about how oral fat is sensed and are relevant to understanding the physiological and pathophysiological processes related to fat intake. PMID- 12761277 TI - PACAP is present in the olfactory system and evokes calcium transients in olfactory receptor neurons. AB - Pituitary adenylate cyclase activating peptide (PACAP), a neuroregulatory peptide, is found in germinative regions of the CNS, including the olfactory bulb, throughout adulthood. We show that 1) PACAP immunoreactivity is also present in the neonatal mouse and adult mouse and rat olfactory epithelium, 2) PACAP expression pattern differs between neonatal and adult mice, and 3) PACAP is produced by olfactory ensheathing cells. PACAP may thus be a key factor in the uniquely supportive role of olfactory ensheathing cells in regeneration of neurons from olfactory epithelium and lesioned spinal cord. Using calcium imaging, we demonstrated physiological responses to PACAP in both neonatal and adult olfactory receptor neurons (ORNs). We propose that PACAP plays an important role in normal turnover of ORNs by providing neurotrophic support during development and regeneration and neuroprotective support of mature neurons. PMID- 12761279 TI - Long-term depression of synaptic inhibition is expressed postsynaptically in the developing auditory system. AB - Inhibitory transmission is critically involved in the functional maturation of neural circuits within the brain. However, the mechanisms involved in its plasticity and development remain poorly understood. At an inhibitory synapse of the developing auditory brain stem, we used whole cell recordings to determine the site of induction and expression of long-term depression (LTD), a robust activity-dependent phenomenon that decreases inhibitory synaptic gain and is postulated to underlie synapse elimination. Recordings were obtained from lateral superior olivary (LSO) neurons, and hyperpolarizing inhibitory potentials were evoked by stimulation of the medial nucleus of the trapezoid body (MNTB). Both postsynaptic glycine and GABAA receptors could independently display LTD when isolated pharmacologically. Focal application of GABA, but not glycine, on the postsynaptic LSO neuron was sufficient to induce depression of the amino acid evoked response, or MNTB-evoked inhibitory postsynaptic potentials. This GABA mediated depression, in the absence of MNTB stimulation, was blocked by a GABAB receptor antagonist. To assess whether a change in neurotransmitter release is associated with the LTD, the polyvalent cation, ruthenium red, was used to increase the frequency of miniature inhibitory synaptic events. Consistent with a postsynaptic locus of expression, we found that the mean amplitude of miniature events decreased after LTD with no change in their frequency of occurrence. Furthermore, there was no change in the paired-pulse ratio or release kinetics of evoked inhibitory responses. Together, these results provide direct evidence that activity-dependent LTD of inhibition has a postsynaptic locus of induction and alteration, and that GABA but not glycine plays a pivotal role. PMID- 12761280 TI - Selective effects of light exposure on distribution of motility in the chick embryo at E18. AB - It is well established that orderly patterns of motor neuron activity, muscle recruitment, and limb movement are generated in chicks during motility by embryonic day (E)9, the midpoint in embryonic development. However, our recent work suggests that some attributes of motility, such as the rhythm of repetitive limb movements and distribution of activity, become less orderly after E9. In this study, we extend these observations by performing continuous force recordings over a 24-h period in ovo at E18 with augmented sampling of synchronized video and electromyogram (EMG) recordings. We report the distribution of three repetitive behaviors, rapid limb movement, respiratory-like movement, and beak clapping, identified in force recordings, and the general distribution of motility. We also test a model recently proposed to account for age-related changes in motility parameters. In the model, we proposed that circadian networks contribute to the age-related changes in distribution of motility. As a first test of this hypothesis, we examine whether light exposure contributes to the variable distribution of motility by comparing motility parameters at E18 for embryos incubated and tested under either a 12-h light/dark cycle or continuous light. Results suggest that exposure to light increases the total amount of activity and hastens the onset of extended respiratory-like movement sequences but does not impact expression of repetitive limb movement or beak clapping at E18. The possible influence of circadian mechanisms on embryonic behavior and insensitivity of repetitive limb movements to light exposure are discussed. PMID- 12761281 TI - Shaker K+ channels contribute early nonlinear amplification to the light response in Drosophila photoreceptors. AB - We describe the contribution of rapidly inactivating Shaker K+ channels to the dynamic membrane properties of Drosophila photoreceptors. Phototransduction was measured in wild-type and Shaker mutant (Sh14) Drosophila photoreceptors by stimulating with white noise-modulated light contrast and recording the resulting intracellular membrane potential fluctuations. A second-order Volterra kernel series was used to characterize the nonlinear dynamic properties of transduction in the two situations. First-order kernels were indistinguishable in wild-type and Sh14 photoreceptors, indicating that the basic light transduction machinery was always intact. However, second-order kernels of Shaker mutants lacked a large, early amplification, indicating a novel role for Shaker K+ channels in amplifying and accelerating the voltage response of wild-type photoreceptors. A cascade model of two nonlinear static components surrounding one linear dynamic component was able to partially reproduce the experimental responses. Parameters obtained by fitting the model to the experimental data supported the hypothesis that normal Shaker K+ channels contribute an early, positive nonlinearity that partially offsets a later attenuating nonlinearity caused by membrane shunting. PMID- 12761282 TI - A comparison of macaque behavior and superior colliculus neuronal activity to predictions from models of two-choice decisions. AB - Recently, models in psychology have been shown capable of accounting for the full range of behavioral data from simple two-choice decision tasks: mean reaction times for correct and error responses, accuracy, and the reaction time distributions for correct and error responses. At the same time, recent data from neural recordings have allowed investigation of the neural systems that implement such decisions. In the experiment presented here, neural recordings were obtained from superior colliculus prelude/buildup cells in two monkeys while they performed a two-choice task that has been used in humans for testing psychological models of the decision process. The best-developed psychological model, the diffusion model, and a competing model, the Poisson counter model, were explicitly fit to the behavioral data. The pattern of activity shown in the prelude/buildup cells, including the point at which response choices were discriminated, was matched by the evidence accumulation process predicted from the diffusion model using the parameters from the fits to the behavioral data but not by the Poisson counter model. These results suggest that prelude/buildup cells in the superior colliculus, or cells in circuits in which the superior colliculus cells participate, implement a diffusion decision process or a variant of the diffusion process. PMID- 12761283 TI - Muscarinic and nicotinic ACh receptor activation differentially mobilize Ca2+ in rat intracardiac ganglion neurons. AB - The origin of intracellular Ca2+ concentration ([Ca2+]i) transients stimulated by nicotinic (nAChR) and muscarinic (mAChR) receptor activation was investigated in fura-2-loaded neonatal rat intracardiac neurons. ACh evoked [Ca2+]i increases that were reduced to approximately 60% of control in the presence of either atropine (1 microM) or mecamylamine (3 microM) and to <20% in the presence of both antagonists. Removal of external Ca2+ reduced ACh-induced responses to 58% of control, which was unchanged in the presence of mecamylamine but reduced to 5% of control by atropine. The nAChR-induced [Ca2+]i response was reduced to 50% by 10 microM ryanodine, whereas the mAChR-induced response was unaffected by ryanodine, suggesting that Ca2+ release from ryanodine-sensitive Ca2+ stores may only contribute to the nAChR-induced [Ca2+]i responses. Perforated-patch whole cell recording at -60 mV shows that the rise in [Ca2+]i is concomitant with slow outward currents on mAChR activation and with rapid inward currents after nAChR activation. In conclusion, different signaling pathways mediate the rise in [Ca2+]i and membrane currents evoked by ACh binding to nicotinic and muscarinic receptors in rat intracardiac neurons. PMID- 12761284 TI - Glycinergic inputs to cardiac vagal neurons in the nucleus ambiguus are inhibited by nociceptin and mu-selective opioids. AB - Most parasympathetic regulation of heart rate originates from preganglionic cardiac vagal neurons within the nucleus ambiguus. Little is known regarding the modulation of glycinergic transmission to these neurons. However, the presence of mu-opioid receptors and opioid-receptor-like (ORL1) receptors within the ambiguus, together with the presence of endogenous ligands for both receptor types in the same area, suggests opioids may modulate synaptic transmission to cardiac vagal neurons. This study therefore examined the effects of endomorphin-1 and endomorphin-2 (the mu-selective endogenous peptides), DAMGO (a synthetic, mu selective agonist), and nociceptin (the ORL1-selective endogenous peptide) on spontaneous glycinergic inhibitory postsynaptic currents (IPSCs) in rat cardiac parasympathetic neurons. All four of the opioids used in this study decreased spontaneous IPSCs. At concentrations of 100 microM, the amplitude of the IPSCs was reduced significantly by nociceptin (-56.6%), DAMGO (-46.5%), endomorphin-1 ( 45.1%), and endomorphin-2 (-26%). IPSC frequency was also significantly reduced by nociceptin (-61.1%), DAMGO (-69.9%), and endomorphin-1 (-40.8%) but not endomorphin-2. Lower concentrations of nociceptin and DAMGO (10-30 microM) also effectively decreased IPSC amplitude and frequency. The inhibitory effects of DAMGO were blocked by d-Phe-Cys-Tyr-d-Trp-Orn-Thr-Pen-Thr-NH2 (C-TOP; 10 microM), a selective mu-receptor antagonist. Neither nociceptin nor DAMGO inhibited the postsynaptic responses evoked by exogenous application of glycine or affected TTX insensitive glycinergic mini-IPSCs. These results indicate that mu-selective opioids and nociceptin act on preceding neurons to decrease glycinergic inputs to cardiac vagal neurons in the nucleus ambiguus. The resulting decrease in glycinergic transmission would increase parasympathetic activity to the heart and may be a mechanism by which opioids induce bradycardia. PMID- 12761285 TI - Biosynthetic regulation and intracellular transport of phosphatidylserine in mammalian cells. AB - In mammalian cells, phosphatidylserine (PtdSer) is synthesized through the action of the endoplasmic reticulum enzymes, PtdSer synthase 1 and 2, and the decarboxylation of PtdSer accounts for the majority of phosphatidylethanolamine (PtdEtn) synthesis. PtdSer decarboxylation for PtdEtn formation occurs in the mitochondria. In addition, the transport of PtdSer from the endoplasmic reticulum to the mitochondria is probably a rate limiting step for PtdEtn synthesis through the decarboxylation pathway. Therefore, the regulation of PtdSer synthesis and its intracellular transport appear to be essential events for the maintenance of normal cellular PtdSer and PtdEtn levels. Here we describe the current understanding of the regulation of PtdSer biosynthesis and the transport of PtdSer from the ER to the mitochondria in mammalian cells. PMID- 12761286 TI - Genome structure and differential expression of two isoforms of a novel PDZ containing myosin (MysPDZ) (Myo18A). AB - We previously cloned a gene for a novel myosin (called MysPDZ) containing a PDZ domain from bone marrow stromal cells. This new myosin is found in humans and classified as one of the class XVIII myosins (Myo18A). Here, we report the hematopoietic cell-specific splicing isoform (MysPDZbeta) in addition to the previously reported isoform (MysPDZalpha). Combined with mouse genome sequence data, the overall genome structure and generation of the two spliced isoforms are deduced. The MysPDZbeta protein lacks a PDZ-domain in the N-terminal region. Studies of the subcellular localization of the two spliced isoforms indicated that MysPDZalpha containing the PDZ domain co-localizes with the ER-Golgi complex, while MysPDZbeta, which lacks the PDZ domain, localizes diffusely in the cytoplasm. These results suggest that the isoforms differ in their subcellular localization and may have different functions in membrane ruffling and membrane traffic pathways. The PDZ-containing spliced isoform (MysPDZalpha) is not expressed in bone marrow hematopoietic cells, whereas MysPDZbeta lacking the PDZ is specifically expressed in most hematopoietic cells. It is noted that neither isoform is expressed in red blood cells. Interestingly, MysPDZalpha was detected in mature but not in immature macrophages, and its level increased after the induction of differentiation of M1 cells, suggesting a functional role of PDZ containing myosin in macrophages. PMID- 12761287 TI - Comparative analysis of yeast PIAS-type SUMO ligases in vivo and in vitro. AB - SUMO/Smt3, a ubiquitin-like modifier, is known to conjugate other proteins and modulate their functions in various processes. Recently, Ull1/Siz1 was discovered as a novel PIAS-type E3 required for septin sumoylation in yeast. We demonstrate here that the second PIAS-type Nfi1/Siz2 is also a SUMO ligase. It interacted with Smt3, SUMO/Smt3 conjugating enzyme Ubc9 and a septin component Cdc3 in the two-hybrid system. The region containing the RING-like domain of Nfi1/Siz2 bound directly to Ubc9 and Cdc3, but not to Smt3. Nfi1/Siz2 stimulated Smt3 conjugation to Cdc3 in vitro. In this in vitro system, Smt3 formed polymeric chains in the presence of higher concentrations of E1 and E2 enzymes. When the lysine(15) residue of Smt3 was substituted with arginine, Smt3 chain-polymerization was abolished. Using this polysumoylation-deficient mutant Smt3, we found that Cdc3 and Nfi1/Siz2 were modified with Smt3 at multiple sites. Finally we found that the C-terminal truncated form of Ull1/Siz1 was mis-localized in vivo, but retained its SUMO ligase activity in vitro. We discuss the regulation of these SUMO ligases in vivo and in vitro. PMID- 12761288 TI - Sustained expression of Fc-fusion cytokine following in vivo electroporation and mouse strain differences in expression levels. AB - We previously demonstrated that cytokine expression following intramuscular gene transfer of a naked plasmid is increased 2 logs by in vivo electroporation, but the relatively low expression levels of the encoded protein is still a limitation for successful gene therapy and gene function studies. We recently reported that the serum viral IL-10 levels achieved by electroporation-mediated intramuscular delivery of pCAGGS-vIL10, a viral IL-10-expressing plasmid, can be further enhanced by modifying the plasmid into an immunoglobulin fusion protein expression plasmid, pCAGGS-vIL10/Fc. Here we examined the applicability of this approach to the expression of an endogenous cytokine, IL-10, in two different inbred mouse strains. We obtained sustained high serum levels of IL-10 in C3H/HeJ mice (C3H), but the level and duration of the gene expression was mouse-strain dependent. Although the serum IL-10 level was also increased by using the IL 10/Fc gene plasmid in C57BL/6 mice (B6), IL-10/Fc and a luciferase reporter showed significantly lower levels in B6 than in C3H mice, and the persistence of pCAGGS-IL10/Fc expression ranged from several days in B6 mice to more than one month in C3H mice. These results suggest that the electroporation-mediated intramuscular delivery of the immunoglobulin fusion protein expression plasmid is simple and very efficient, but mouse strain differences in transgene expression should be taken into consideration in its use. PMID- 12761289 TI - Identification of a novel, embryonal carcinoma cell-associated molecule, nucling, that is up-regulated during cardiac muscle differentiation. AB - EC cells are characterized by their potent capacity to differentiate into several cell types, such as mesoderm-like cells, endoderm-like cells, or ectoderm-like cells. By subtracting the mRNAs expressed by one EC cell clone, F9 cells, with the mRNAs expressed by another EC cell clone, P19 cells, we identified six novel genes that are expressed selectively by F9 cells. One of these genes (Nucling) encodes a polypeptide of 1411 amino acids containing an ankyrin repeat, aspartyl protease motif, a leucine zipper motif, and two t-SNARE coiled-coil domains. Northern blot analyses revealed the Nucling mRNA to be detected predominantly in heart, liver, kidney and testis, but not in brain or spleen. Immunostaining analyses revealed a unique feature of Nucling that the transiently expressed protein forms aggregates exclusively around nuclear membranes. Moreover, the expression level of the Nucling gene transcript increases progressively during the early developmental stages in mice, and specifically at cardiomuscular differentiation in vitro and in vivo. These results suggest that Nucling may play some role in the gene regulation of cell differentiation during embryonal development. PMID- 12761290 TI - Isolation of small agranular synaptic vesicles of rat brain by gel filtration chromatography. AB - I attempted to isolate synaptic vesicles by gel filtration. The rat brain synaptic vesicles in a synaptosomal lysate were collected by ammonium sulfate salting-out and fractionated on a Sephacryl S-500 with a mean exclusion size of 200 nm. Peak I at the void volume contained large vesicular membranes and coated vesicles besides synaptic vesicles; Peak II consisted almost entirely of small agranular synaptic vesicles of 40-50 nm diameter; and Peak III comprised soluble proteins. Western blotting revealed that components of 72 kDa in peaks I and II reacted with an anti-H(+)-ATPase A-subunit antibody [Moriyama et al. (1995) FEBS Lett. 367, 233-236]. When examined for Mg(2+)-ATPase activity, peak I showed specific activity of 4.52 ( micromol ATP hydrolyzed/mg protein/30 min), while that of peak II was as low as 0.22. As estimated from the inhibition by bafilomycin A(1) [Bowman et al. (1988) PROC: Natl. Acad. Sci. USA 85, 7972-7976], the percentage of H(+)-ATPase as to total Mg(2+)-ATPase, 18-22%, was unchanged, indicating no accumulation of the H(+)-ATPase in peak II even on the chromatography. In brief, the small agranular synaptic vesicles in peak II showed little or no Mg(2+)-ATPase activity, although they reacted with the H(+)-ATPase antibody. The reason for this is obscure. Mg(2+)-ATPase might not be a constituent of small agranular synaptic vesicles of rat brain. PMID- 12761291 TI - Substrate shape specificity of E coli RNase P ribozyme is dependent on the concentration of magnesium ion. AB - The bacterial RNase P ribozyme can accept a hairpin RNA with CCA-3' tag sequence as well as a cloverleaf pre-tRNA as substrate in vitro, but the details are not known. By switching tRNA structure using an antisense guide DNA technique, we examined the Escherichia coli RNase P ribozyme specificity for substrate RNA of a given shape. Analysis of the RNase P reaction with various concentrations of magnesium ion revealed that the ribozyme cleaved only the cloverleaf RNA at below 10 mM magnesium ion. At 10 mM magnesium ion or more, the ribozyme also cleaved a hairpin RNA with a CCA-3' tag sequence. At above 20 mM magnesium ion, cleavage site wobbling by the enzyme in tRNA-derived hairpin occurred, and the substrate specificity of the enzyme became broader. Additional studies using another hairpin substrate demonstrated the same tendency. Our data strongly suggest that raising the concentration of metal ion induces a conformational change in the RNA enzyme. PMID- 12761292 TI - Characterization of rat gastric mucins using a monoclonal antibody, RGM23, recognizing surface mucous cell-type mucins. AB - A novel anti-mucin monoclonal antibody (mAb), designated RGM23, was developed against mucin purified from rat gastric mucosa. RGM23 reacted with the mucin attached to the ELISA well. The reactivity was lost by trypsin treatment, but not by periodate oxidation, indicating that RGM23 recognizes the peptide moiety of the mucin molecule. Histochemical study showed that RGM23 stained the corpus and antral surface mucosa of rat stomach, but not their glandular mucosa, nor duodenal, small intestinal or large intestinal mucosa. The area stained with RGM23 was coincident with that stained with 45M1, a mAb reacting with MUC5AC mucin. Examination of the mucin subunits extracted from rat stomach by Sepharose CL-4B and Q-Sepharose chromatography and CsTFA equilibrium centrifugation showed that RGM23 reacted with the surface mucous cell-type mucins that were stained with periodate-Schiff (PAS) and reacted with mAb RGM21. The gastric gland-type mucin, which reacted with mAb HIK1083, did not react with RGM23. On Q-Sepharose chromatography, a part of the RGM21-reactive mucins was only faintly stained with PAS and did not react with RGM23. The results together indicated that RGM23 probably reacted with the rat MUC5AC (rMuc5AC) mucin present in the surface mucosa of the stomach, and that the surface mucosal cells in rat stomach may contain mucin bearing non-rMuc5AC core protein in addition to rMuc5AC mucins. PMID- 12761293 TI - Hybrid respiration in the denitrifying mitochondria of Fusarium oxysporum. AB - Induction of the mitochondrial nitrate-respiration (denitrification) system of the fungus Fusarium oxysporum requires the supply of low levels of oxygen (O(2)). Here we show that O(2) and nitrate (NO(3)(-)) respiration function simultaneously in the mitochondria of fungal cells incubated under hypoxic, denitrifying conditions in which both O(2) and NO(3)(-) act as the terminal electron acceptors. The NO(3)(-) and nitrite (NO(2)(-)) reductases involved in fungal denitrification share the mitochondrial respiratory chain with cytochrome oxidase. F. oxysporum cytochrome c(549) can serve as an electron donor for both NO(2)(-) reductase and cytochrome oxidase. We are the first to demonstrate hybrid respiration in respiring eukaryotic mitochondria. PMID- 12761294 TI - Crystal structures of beta-amylase from Bacillus cereus var mycoides in complexes with substrate analogs and affinity-labeling reagents. AB - The crystal structures of beta-amylase from Bacillus cereus var. mycoides in complexes with five inhibitors were solved. The inhibitors used were three substrate analogs, i.e. glucose, maltose (product), and a synthesized compound, O alpha-D-glucopyranosyl-(1-->4)-O-alpha-D-glucopyranosyl-(1-->4)-D-xylopyranose (GGX), and two affinity-labeling reagents with an epoxy alkyl group at the reducing end of glucose. For all inhibitors, one molecule was bound at the active site cleft and the non-reducing end glucose of the four inhibitors except GGX was located at subsite 1, accompanied by a large conformational change of the flexible loop (residues 93-97), which covered the bound inhibitor. In addition, another molecule of maltose or GGX was bound about 30 A away from the active site. A large movement of residues 330 and 331 around subsite 3 was also observed upon the binding of GGX at subsites 3 to 5. Two affinity-labeling reagents, alpha EPG and alpha-EBG, were covalently bound to a catalytic residue (Glu-172). A substrate recognition mechanism for the beta-amylase was discussed based on the modes of binding of these inhibitors in the active site cleft. PMID- 12761296 TI - A novel screening system for self-mRNA targeting proteins. AB - Here we describe the application of an in vitro translation system for genetic screening, to identify RNA-binding proteins that bind to their own mRNAs. It is a relatively novel system designed using an advanced cell-free translation system reconstructed with purified translational components. Due to the absence of nucleases and proteases, the complex of mRNA and nascent polypeptide synthesized in this system is expected to exhibit high stability ensuring the following efficient selection toward the protein. Escherichia coli ribosomal protein S15, which is known to bind to its own mRNA, was employed as a model molecule to evaluate the system. Wild-type S15 mRNA specifically isolated from a mutant mRNA lacking the secondary structure responsible for binding the S15 protein accumulated markedly after several rounds of selection-amplification. The success of this selection demonstrates the potentiality of the systematic screening of self-mRNA targeting proteins through direct and functional selection. This strategy as a method to identify peptides or proteins that bind to their own mRNAs, is of general interest and has different potential applications, such as, the identification of new regulatory proteins or peptide motifs for RNA recognition, the study of self-mRNA-protein interactions, etc. PMID- 12761295 TI - Transcriptional, functional and cytochemical analyses of the veg gene in Bacillus subtilis. AB - A Bacillus subtilis veg mutant exhibited a small reduction of absorbance, a large reduction of hexosamine release, and slow dipicolinic acid release from spores during germination with L-alanine as a germinant. But veg spores exhibited normal resistance to chloroform, 2-propanol, lysozyme, and heat. SDS-polyacrylamide gel electrophoresis of spore coat proteins revealed no difference in coat proteins between the wild type and the veg mutant. Northern and veg-lacZ fusion analyses indicated that the veg gene is transcribed in both the vegetative growth and sporulation phases, and primer extension analysis indicated an identical transcriptional start point in both phases. The upstream sequence suggests that veg is transcribed by Esigma(A) RNA polymerase. Veg-GFP fusion protein was detected for vegetative cells and spores, but the fluorescence of mother cells disappeared completely in the late sporulation phase. Decoated spores containing Veg-GFP exhibited a strong green fluorescence in the core, but much weaker fluorescence in the cortex. PMID- 12761297 TI - Identification of a novel tissue-specific transcriptional activator FESTA as a protein that interacts with the transcription elongation factor S-II. AB - Transcription elongation factor S-II was originally purified as a specific stimulator of transcription by RNA polymerase II. Recent studies suggest that S II participates in gene-specific transcriptional activation in vivo, despite the fact that it directly binds RNA polymerase II and does not recognize specific DNA sequences. In this study, under the hypothesis that S-II requires co-factors to regulate the expression of specific-genes in vivo, we searched for factors that directly interact with S-II using a yeast two-hybrid system, and isolated a novel nuclear protein, FESTA. FESTA is expressed specifically in kidney and spleen, supporting our notion that S-II participates in gene-specific regulation. Two mRNA isoforms of FESTA encoding proteins with different sizes were identified and named FESTA-S and FESTA-L. FESTA contains a serine-rich region and a C-terminal tail that are highly similar to those of the ELL-associated factor EAF1. Reporter gene assays indicated that both GAL4-FESTA-S and GAL4-FESTA-L fusion proteins have trans-activating ability. Furthermore, deletion of the C-terminal tail of FESTA dramatically reduced its trans-activating ability and abolished its interaction with S-II. This study is the first report of a transcriptional activator that directly interacts with S-II and contains a transcriptional activation domain that cooperates with S-II via direct interaction. PMID- 12761298 TI - Nitric oxide synthase induction, cGMP elevation, and biopterin synthesis in vascular smooth muscle cells stimulated with interleukin-1beta in hypoxia. AB - In cultured rat vascular smooth muscle cells (VSMC), inducible nitric oxide synthase (iNOS) expression evoked by interleukin-1beta (IL-1beta) or tumor necrosis factor-alpha was greatly enhanced in hypoxia (2% O(2)), compared to in normoxia. In contrast, iNOS induction by interferon-gamma, lipopolysaccharide or their combination was barely influenced by hypoxia. These results indicate that iNOS induction is regulated by hypoxia in different manners, depending on the stimuli in VSMC. Nitric oxide (NO) production in response to stimulation with interferon-gamma plus lipopolysaccharide was significantly decreased in hypoxia, due to a decrease in the concentration of O(2) as a substrate. In contrast, the level of NO production in hypoxia was almost the same as that in normoxia when the cells were stimulated by IL-1beta. In addition, cGMP increased in response to IL-1beta in hypoxia to a level comparable to that in normoxia. Thus, it seems that the IL-1beta-induced expression of iNOS is up-regulated in hypoxia to compensate for a decrease in the enzyme activity due to the lower availability of O(2) as a substrate, and consequently a sufficient amount of NO is produced to elevate cGMP to an adequate level. In addition, the IL-1beta-induced synthesis of tetrahydrobiopterin, a cofactor for iNOS, was also greatly stimulated by hypoxia in VSMC. PMID- 12761299 TI - Compositional changes in RNA, DNA and proteins for bacterial adaptation to higher and lower temperatures. AB - It is known that in thermophiles the G+C content of ribosomal RNA linearly correlates with growth temperature, while that of genomic DNA does not. Although the G+C contents (singlet) of the genomic DNAs of thermophiles and methophiles do not differ significantly, the dinucleotide (doublet) compositions of the two bacterial groups clearly do. The average amino acid compositions of proteins of the two groups are also distinct. Based on these facts, we here analyzed the DNA and protein compositions of various bacteria in terms of the optimal growth temperature (OGT). Regression analyses of the sequence data for thermophilic, mesophilic and psychrophilic bacteria revealed good linear relationships between OGT and the dinucleotide compositions of DNA, and between OGT and the amino acid compositions of proteins. Together with the above-mentioned linear relationship between ribosomal RNA and OGT, the DNA and protein compositions can be regarded as thermostability measures for RNA, DNA and proteins, covering a wide range of temperatures. Both the DNA and proteins of psychrophiles apparently exhibit characteristics diametrically opposite to those of thermophiles. The physicochemical parameters of dinucleotides suggested that supercoiling of DNA is relevant to its thermostability. Protein stability in thermophiles is realized primarily through global changes that increase charged residues (i.e., Glu, Arg, and Lys) on the molecular surface of all proteins. This kind of global change is attainable through a change in the amino acid composition coupled with alterations in the DNA base composition. The general strategies of thermophiles and psychrophiles for adaptation to higher and lower temperatures, respectively, that are suggested by the present study are discussed. PMID- 12761300 TI - Role of the yeast VAP homolog, Scs2p, in INO1 expression and phospholipid metabolism. AB - In the yeast Saccharomyces cerevisiae, the expression of phospholipid biosynthetic genes, including the INO1 gene (encoding inositol-1-phosphate synthase), is coordinately regulated by a cis-acting transcriptional element, UAS(INO) (inositol-sensitive upstream activating sequence). For this paper we studied the effect of SCS2 disruption on INO1 expression. SCS2 encodes a type II membrane protein and its deletion leads to inositol auxotrophy at temperatures above 34 degrees C. We found that the expression of the INO1 gene was reduced in the scs2Delta strain even when the cells were cultured under derepressing conditions for INO1 expression. However, the beta-galactosidase gene fused with the INO1 promoter region was expressed normally in the scs2Delta strain. The phospholipid composition of scs2Delta cells was not dramatically changed compared with wild-type cells at 28 degrees C, but the phosphatidylinositol level was reduced in scs2Delta cells cultured at 34 degrees C. In addition, elevated phosphatidylcholine synthesis through the CDP-choline pathway was observed in the scs2Delta strain, and the disruption of genes involved in the CDP-choline pathway rescued the INO1 expression defect of the scs2Delta strain. These results indicate that Scs2p can contribute to coordinated phospholipid metabolism including INO1 expression by regulating phosphatidylcholine synthesis through the CDP-choline pathway. PMID- 12761301 TI - The gene encoding muscle-type carnitine palmitoyltransferase I: comparison of the 5'-upstream region of human and rodent genes. AB - Muscle-type carnitine palmitoyltransferase I (M-CPTI) is the key enzyme for fatty acid beta-oxidation in heart and skeletal muscles and in adipose tissue. So far, M-CPTI mRNA has been detected in white adipocytes from epididymal fat pads of rats and humans, but not in mouse adipocytes. To characterize the gene expression of M-CPTI in mice, we isolated the genomic DNA encoding mouse M-CPTI and determined its transcription initiation site. As a result, the mouse M-CPTI gene seemed to have multiple initiation sites, as in the case of the rat and human genes. Furthermore, the conserved nucleotide sequence of the response element for peroxisome proliferators was shown to exist in the upstream of the mouse gene as in that of the rat and human genes. From these observations, we suggest that the anomalous expression of M-CPTI in mouse adipocytes reported previously may be regulated by factors other than peroxisome proliferators. Previously, we reported that there were transcripts containing regions of both CK/EK-beta and M-CPTI genes in humans. In this study, we found that such transcripts also exist in rodents and that the amounts of the transcripts containing regions of both of these genes did not depend on the expression level of CK/EK-beta. PMID- 12761302 TI - Drosophila DSP1 and rat HMGB1 have equivalent DNA binding properties and share a similar secondary fold. AB - The protein DSP1 belongs to the group of HMG-box proteins, which share the common structural feature of the HMG-box. This approximately 80 amino acid long motif binds DNA via the minor groove. DSP1 was discovered as a transcriptional co repressor of Dorsal in Drosophila melanogaster and then was shown to participate to the remodeling of chromatin. By means of sequence alignment and gene organization, DSP1 was classified as the fly homologue of the vertebrate proteins HMGB1/2. DSP1 contains two HMG boxes flanked by two glutamine-rich domains at the N-terminus. In addition, the HMG domain of DSP1 displays two differences in its primary sequence as compared to the vertebrate HMGB1: a shorter acidic tail and a linker between the two boxes longer by 6 amino acids. By comparing several functional parameters of DSP1 with those of HMGB1, the present study establishes the functional equivalence of both proteins in terms of DNA recognition. The major structural difference between the two proteins, the glutamine-rich N terminal tail of DSP1, which does not exist in HMGB1, did not interfere with any of the studied DNA-binding properties of the proteins. PMID- 12761303 TI - Molecular action mode of Hippospongic acid A, an inhibitor of gastrulation of starfish embryos. AB - Hippospongic acid A (HA-A) is a novel natural triterpene metabolite that exhibits inhibitory activity against the gastrulation of starfish embryos isolated from a marine sponge, Hippospongia sp. We succeeded in chemically synthesizing the natural enantiomer and the racemate HA-A. In this study, we examined its action mode in vitro. HA-A was a rare compound that could selectively but uniformly inhibit the activities of all the vertebrate DNA polymerases tested such as alpha, beta, delta, epsilon, eta, kappa, and lambda, in the IC(50) range of 5.9 17.6 microM, and interestingly also those of human DNA topoisomerases I and II (IC(50) = 15-25 microM). HA-A exhibited no inhibitory effect on DNA polymerases from insects, plants and prokaryotes, or on many other DNA metabolic enzymes. HA A was an inhibitor specific to DNA polymerases and DNA topoisomerases from vertebrates, but not selective as to a subclass species among the enzymes. Since DNA polymerase beta is the smallest, we used it to analyze the biochemical relationship with HA-A. Biochemical, BIAcore and computer modeling analyses demonstrated that HA-A bound selectively to the N-terminal 8 kDa DNA template binding domain of DNA polymerase beta, and HA-A inhibited the ssDNA binding activity. HA-A could prevent the growth of NUGC-3 cancer cells at both the G1 and G2/M phases, and induce apoptosis in the cells. The LD(50) value was 9.5 microM, i.e. in the same range as for the enzyme inhibition. Therefore, we concluded that one molecular basis of the gastrulation of starfish embryos is a process that requires DNA polymerases and DNA topoisomerases, and subsequently the gastrulation was inhibited by HA-A. We also discussed the in vivo role of HA-A. PMID- 12761304 TI - Xylose reductase from the Basidiomycete fungus Cryptococcus flavus: purification, steady-state kinetic characterization, and detailed analysis of the substrate binding pocket using structure-activity relationships. AB - Xylose reductase has been purified to apparent homogeneity from cell extracts of the fungus Cryptococcus flavus grown on D-xylose as carbon source. The enzyme, the first of its kind from the phylum Basidiomycota, is a functional dimer composed of identical subunits of 35.3 kDa mass and requires NADP(H) for activity. Steady-state kinetic parameters for the reaction, D-xylose + NADPH + H(+)<--> xylitol + NADP(+), have been obtained at pH 7.0 and 25 degrees C. The catalytic efficiency for reduction of D-xylose is 150 times that for oxidation of xylitol. This and the 3-fold tighter binding of NADPH than NADP(+) indicate that the enzyme is primed for unidirectional metabolic function in microbial physiology. Kinetic analysis of enzymic reduction of aldehyde substrates differing in hydrophobic and hydrogen bonding capabilities with binary enzyme NADPH complex has been used to characterize the substrate-binding pocket of xylose reductase. Total transition state stabilization energy derived from bonding with non-reacting sugar hydroxyls is approximately 15 kJ/mol, with a major contribution of 5-8 kJ/mol made by interactions with the C-2(R) hydroxy group. The aldehyde binding site is approximately 1.2 times more hydrophobic than n-octanol and can accommodate linear alkyl chains of 1 as a result of noncompressible vessels and the estimated ABI correctly reflected PVD. In the second case, the ABI was near normal because of a patent peroneal artery and our estimated ABI was abnormally low because our estimation method failed to factor in the peroneal artery. CONCLUSIONS: The proposed angiography scoring system reproducibly estimated flow reductions to distal tissue beds. The authors plan to use this system as a research tool for evaluating new methods of assessing and treating PVD. PMID- 12761309 TI - Prevalence and treatment of cephalic arch stenosis in dysfunctional autogenous hemodialysis fistulas. AB - PURPOSE: Cephalic arch stenosis (CAS) is a recently recognized cause of dysfunction in autogenous hemodialysis fistulas. The prevalence of this lesion among dysfunctional autogenous fistulas is described, as are outcomes after percutaneous therapy. MATERIALS AND METHODS: A cohort of 177 dysfunctional autogenous fistulas treated over a 48-month period was retrospectively analyzed for the presence of CAS. Of these, 116 (66%) were radiocephalic fistulas and 61 (34%) were brachiocephalic fistulas. CAS was identified in 26 fistulas among 24 patients. Fifty dilations and three stent placements in the cephalic arch were performed. Surveillance was conducted after percutaneous therapy by means of ultrasound dilution technique and measurement of dialysis flow rates. Patency rates were estimated with use of the Kaplan-Meier method. No patients were lost to follow-up. RESULTS: The prevalence of CAS was 15% (26 of 177). There was a significant difference in the prevalence of CAS between brachiocephalic and radiocephalic fistulas (39% vs 2%; P <.001). High-pressure noncompliant balloon catheters were required in 29 of 50 dilations (58%) to efface the lesion. Primary patency rates (+/-SE) at 3, 6, and 12 months were 76% +/- 8, 42% +/- 10, and 23% +/- 9, respectively. Primary assisted patency rates (+/-SE) at 3, 6, and 12 months were 96% +/- 4, 83% +/- 8, and 75% +/- 10. Complications occurred in three cases (6%). A major complication with rupture of the cephalic arch resulted in thrombosis and fistula loss (n = 1); two minor complications of cephalic arch rupture were salvaged with placement of a Wallstent (n = 1) or prolonged balloon inflation (n = 1). CONCLUSIONS: CAS is common among failing brachiocephalic arteriovenous fistulas. With aggressive percutaneous intervention and surveillance, favorable primary assisted patency rates can be achieved. PMID- 12761310 TI - Radiofrequency ablation of hepatocellular carcinoma with a saline solution perfusion device: a pilot study. AB - PURPOSE: The majority of patients with hepatocellular carcinoma (HCC) cannot be treated with surgery. This study evaluated the treatment of patients with radiofrequency ablation (RFA) with use of needle applicators perfused with isotonic saline solution. MATERIALS AND METHODS: Twenty patients with a maximum of three HCCs as large as 60 mm and a contraindication to partial liver resection or orthotopic liver transplantation were enrolled in the study. They were treated with ultrasound-guided RFA with use of perfused needle applicators and followed with sonography and computed tomography. RESULTS: Twenty patients (14 men, six women) with a total of 29 HCCs were treated with RFA. In total, 56 RFA treatments were performed. Mean HCC size was 31 mm (range, 10-60 mm). Two cases of self limiting bleeding requiring transfusion of blood products took place. Complete response (CR) was achieved in 85% of patients (17 of 20). Partial response occurred in three tumors 45 mm in diameter or larger. There was a total of six local recurrences in five patients (25% of patients, 21% of tumors). Six patients (30%) exhibited distant recurrence, two of whom also showed local recurrence. In three patients, repeat treatment led to renewed CR. After a median follow-up of 445 days (range, 114-1,071 days), 12 patients (71% of the 17 patients with initial CR) still showed CR. Twelve patients (60% of all patients, 71% of those with initial CR) survived. Three of the eight patients who died were free of viable tumor at the time of death. CONCLUSIONS: RFA with use of perfused needle applicators shows promise as an effective method for treating inoperable HCC. The most frequent complication was bleeding. PMID- 12761311 TI - Factors affecting diagnostic accuracy of CT-guided coaxial cutting needle lung biopsy: retrospective analysis of 631 procedures. AB - PURPOSE: To analyze variables affecting diagnostic accuracy of computed tomography (CT)-guided percutaneous coaxial cutting needle biopsy of lung lesions. MATERIALS AND METHODS: A retrospective analysis of factors affecting diagnostic accuracy of CT-guided percutaneous coaxial cutting needle lung biopsy was performed in 631 consecutive procedures with confirmed final diagnoses. Benign and malignant needle biopsy results were cross-examined with correct and incorrect final outcomes to determine diagnostic accuracy. Factors affecting diagnostic accuracy were determined by multivariate logistic regression analysis of variables thought to affect diagnostic accuracy. A P value less than 0.05 was interpreted as statistically significant. RESULTS: The overall diagnostic accuracy of CT-guided percutaneous coaxial cutting needle biopsy of lung lesions was 95% (95% CI: 92.7%-96.2%). Sensitivity was 93%, specificity 98%, negative predictive value 6%, positive predictive value 99%, false-positive rate 0.7%, and false-negative rate 15%. The factors affecting diagnostic accuracy were final diagnoses (benign, 86%; malignant, 99%; chi(2) test, P < 0.001) and lesion size (lesions <1.5 cm, 84%; lesions 1.5-5.0 cm, 96%; lesions >5 cm, 93%; chi(2) test, P = 0.06). CONCLUSION: Benign lung lesions, lung lesions smaller than 1.5 cm (which pose technical difficulty), and lung lesions larger than 5 cm (which are associated with a higher necrosis rate) affect diagnostic accuracy of CT-guided percutaneous coaxial cutting needle biopsy of lung lesions. PMID- 12761312 TI - US-guided catheter drainage of postoperative head and neck fluid collections. AB - PURPOSE: To analyze results of percutaneous catheter drainage of large fluid collections in the head and neck region noted in the immediate postoperative period. MATERIALS AND METHODS: Thirty-four consecutive patients with 41 large fluid collections in the head and neck detected 7-10 days after oncologic surgery underwent percutaneous catheter drainage. There were 29 men and five women, with a mean age of 52.2 years (95% CI: 47.8-56.7). Each patient had a surgical drain placed in the posterior neck triangle; three patients had low-grade fever and six had diabetes at the time of percutaneous catheter drainage. Conventional management consisting of serial needle aspirations at bedside followed by pressure dressing failed in 15 of 34 patients (44%). Ultrasound-guided drainage was performed and catheters were connected to vacuum balls for continuous suction drainage. RESULTS: The average fluid volume estimated by sonographic measurement was 84 cm(3) (95% CI: 57-112 cm(3)). The fluid content was serosanguinous in 46% (19 of 41), uncoagulated blood in 32% (13 of 41), pus in 15% (six of 41), and saliva in 7% (three of 41). The mean duration of catheter drainage was nine days (95% CI: 7-10 d) and mean fluid drainage was 287 mL (95% CI: 188-387 mL). Ninety one percent of patients (31 of 34) were successfully treated with initial catheter drainage. Three patients had recurrent fluid collections at the same locations: two were treated with repeated catheter insertions and one required a limited open drainage. No complication related to catheter drainage was noted at 6-month follow-up. CONCLUSION: Percutaneous catheter drainage is effective for large fluid collections in the head and neck region noted in the immediate postoperative period irrespective of contents. PMID- 12761313 TI - Percutaneous abscess drainage: use of related radiology services and associated economic impact on a radiology practice. AB - PURPOSE: To evaluate the impact of percutaneous abscess drainage on the usage and professional value of subsequent services provided by a radiology practice. MATERIALS AND METHODS: Percutaneous abscess drainage was selected as a marker interventional radiology procedure because of its pervasiveness and ease of identification of related services. Billing records were reviewed for 48 consecutive patients who underwent abscess drainage during a 9-month period. Current procedural terminology (CPT) codes for all radiology services during the subsequent 90 days were analyzed to identify those related to the initial drainage procedure. Professional relative value unit (RVU) impact was calculated. RESULTS: Initial abscess drainage services were identified by 2.6 +/- 1.2 CPT codes, but patients underwent 13.4 +/- 10.7 related radiology services during the subsequent 90 days. The professional RVU impact of subsequent services was 64% higher than that of initial procedures: initial drainage services accounted for 11.5 +/- 5.1 RVUs and all subsequent related radiology services accounted for 18.9 +/- 16.8 RVUs (P =.0042). Of those, additional interventional radiology procedures amounted to 10.7 +/- 12.8 RVUs, diagnostic radiology services 4.7 +/- 4.6 RVUs, and evaluation and management services 3.5 +/- 2.9 RVUs. CONCLUSION: Basic interventional radiology services may result in far more economic impact on radiology practices than initial direct procedure analyses suggest. For percutaneous abscess drainage, the professional RVU impact of subsequent services exceeds that of the initial procedure by 64%. Practices negotiating capitated contracts for interventional services need to consider the high value of such related services. PMID- 12761315 TI - Comparison of various cerebral protection devices used for carotid artery stent placement: an in vitro experiment. AB - PURPOSE: To compare the effectiveness of five basic cerebral protection devices designed for carotid angioplasty in an in-vitro bench-top model. MATERIALS AND METHODS: Simulation of embolization from carotid angioplasty (n = 180) was performed with polyvinyl alcohol (PVA) particles (Contour; 150-1,000- micro m) in an open flow model simulating the aortic arch with a carotid bifurcation made from elastic silicone tubes. Particles (150-250 micro m [small], 355-500 micro m [medium], 710-1,000 micro m [large]; 5 mg each) were injected separately into the internal carotid artery (ICA) proximal to the placed protection device. Five devices were tested: the Angioguard (AG), Filter Wire EX (EX(A), EX(B)), Trap, Neuroshield (NS), and GuardWire Plus (GW). Particles getting past the protection device or flowing into the external carotid artery (ECA) were caught in a filter at the end of both arteries and their weight was determined. RESULTS: For small, medium, and large particles, the lowest weight of emboli in the effluent of the ICA was obtained with the NS (0.28 mg, 0.18 mg, and 0.07 mg, respectively; P <.001 compared to all other devices except the GW for small particles only). The GW had the highest embolization rate into the ECA for all particle sizes. When combining the particle weights for the different protection devices, the NS showed the lowest weight of emboli into the ICA filter (0.53 mg/3.5%; P <.001 compared to all [>1.1 mg/7.0%]). The GW revealed the highest weight of emboli into the ECA (1.2 mg/7.6%; P <.001 compared to all [<0.59 mg/0.09%]). Effectiveness of the EX(B) device was enhanced when circumferentially deployed under direct view (EX(B), 0.39 mg/2.58%; EX(A), 1.18 mg/7.81%; P <.001). CONCLUSIONS: In vitro, none of the tested devices or modifications has the ability to prevent embolization completely. An occlusion balloon leads to increased embolization into the ECA. The effectiveness of the EX might be enhanced with design improvements. During this evaluation, the NS was most effective for preventing PVA particle embolization of the three different particle groups within this in-vitro model. PMID- 12761314 TI - Heparin-coated balloon-expandable stent reduces intimal hyperplasia in the iliac artery in baboons. AB - PURPOSE: To examine the effect of heparin-coated balloon-expandable iliac stent placement on intimal hyperplasia in a baboon model. MATERIALS AND METHODS: Balloon-expandable (Palmaz-Schatz) stents were placed in bilateral common iliac arteries in 20 male baboons (mean weight: 8.8 kg). In each animal, a heparin coated iliac stent was placed on one side and the contralateral iliac artery received an uncoated stent that served as a control. The iliac artery stents were harvested at 30 days (n = 10) and 90 days (n = 10). Arteriography was performed to assess iliac patency and intravascular ultrasonography was used to determine neointimal and luminal areas. Histologic and morphometric analysis and scanning electron microscopy were performed in the stent-implanted iliac arteries. RESULTS: One animal was excluded in the 30-day group because of premature death. In the remaining nine surviving animals, seven control stents (78%) and nine heparin-coated stents (100%) remained patent. Morphometric analysis showed that the iliac arteries with heparin-coated stents had larger luminal areas (17%, P <.05), less neointimal area (26%, P <.05), lower neointima-to-media ratios (32%, P <.05), and equivalent medial areas (P =.92) compared to the control group at 30 days. In contrast, all control and heparin-coated stents were patent (100%) in the 90-day group. In that group, the heparin-coated stent group had less neointimal area (28%, P <.05), lower neointima-to-media ratios (42%, P <.05), and equivalent medial area (P =.92) and luminal area (P =.07) compared to the control group. CONCLUSIONS: The data demonstrate that heparin-coated balloon-expandable stents reduce intimal hyperplasia after iliac artery stent placement. This approach may represent a useful strategy for prolonging luminal patency after iliac stent placement. PMID- 12761316 TI - In vitro study of the compatibility of tris-acryl gelatin microspheres with various chemotherapeutic agents. AB - PURPOSE: To evaluate the in-vitro effects of chemotherapeutic agents on the physical structure of tris-acryl gelatin microspheres. MATERIALS AND METHODS: Microspheres (Embospheres) were mixed in vitro with carboplatin, mitomycin C, 5 fluorouracil, or pirarubicin as experimental samples and with distilled water as control samples and kept for 24 hours at 37 degrees C. The microspheres used measured 100-300 micro m and 700-900 micro m in diameter. Microsphere morphology, including appearance, overall shape, smoothness of surface, and thickness of gelatin coating, was examined with use of a microscope equipped with a cold light. Microsphere dimensions including larger diameter, smaller diameter, and surface area were measured by granulometry. Microsphere geometry was evaluated by calculating a sphericity index and surface regularity index. Qualitative and quantitative variables, respectively, were analyzed with the two-sided Fisher exact test and Wilcoxon signed-rank test for paired values, with a significance level of 0.05. RESULTS: No broken microspheres or microscopic degradations in the appearance, overall shape, smoothness of surface, or thickness of gelatin coating of any microspheres were observed. The respective distributions of large and small diameters, surface area, sphericity index, or surface regularity index of the microspheres were not significantly different between the experimental samples containing carboplatin, mitomycin C, 5-fluorouracil, or pirarubicin and the control sample of microspheres with similar diameters. CONCLUSION: Embosphere microspheres can be mixed with carboplatin, mitomycin C, 5-fluorouracil, or pirarubicin for chemoembolization therapy without any risk of damaging their morphology, dimensions, or geometric characteristics. PMID- 12761317 TI - Percutaneous chemical ablation of desmoid tumors. AB - Desmoid tumors are rare fibroblastic neoplasms characterized by highly aggressive local invasion and high recurrence rates after surgical excision. Responses to chemotherapy and radiation therapy are uncommon. The use of percutaneous chemical ablation performed under imaging guidance is described in two patients with recurrent and/or unresectable disease. A complete response was achieved in one patient and a partial response was achieved in the other patient; these responses were sustained during 2 years of follow-up. This technique may hold promise in the treatment of these challenging lesions. PMID- 12761318 TI - Detachable balloon-modified reducing stent to treat hepatic insufficiency after transjugular intrahepatic portosystemic shunt creation. AB - A patient developed severe hepatic insufficiency after creation of a transjugular intrahepatic portosystemic shunt, which was treated unsuccessfully with a constrained Wallstent ("reducing stent"). After a failed attempt at occluding the shunt with an occlusion balloon, a detachable balloon was placed at the portal end of the constrained stent. This balloon-modified reducing stent reduced flow while maintaining shunt patency, and the patient's hepatic insufficiency improved. She went on to undergo successful liver transplantation several months later. PMID- 12761319 TI - Withdrawal of implanted port-catheter for hepatic arterial infusion chemotherapy with fixed catheter tip technique. AB - The purpose of the present study is to evaluate the feasibility of a method developed to withdraw a port-catheter system that had been implanted with use of the fixed catheter tip technique. Withdrawal of an implanted catheter was required in four patients with advanced liver cancer in whom port-catheter systems had been implanted for performance of repeated hepatic arterial infusion. In all patients, port-catheter systems were successfully removed without complications. In conclusion, an implanted port-catheter system can be removed even when implanted with the fixed catheter tip technique. PMID- 12761320 TI - Uterine arterial embolization for the treatment of diffuse leiomyomatosis. AB - Diffuse leiomyomatosis of the uterus is an extremely rare condition and presents as a diffuse involvement of the myometrium by numerous leiomyomata. Despite patients being in only the third or fourth decades of life, all previously reported patients with this condition have undergone hysterectomy because myomectomy is impossible for this condition. Uterine arterial embolization (UAE) successfully reduced the uterine volume of diffuse leiomyomatosis without major complications in the case presented herein, and the symptoms improved remarkably. Enhanced magnetic resonance imaging after intravenous injection of gadolinium demonstrated global infarction of the vast majority of nodules. PMID- 12761321 TI - SIR 2003 film panel case 1: Behcet vasculitis. PMID- 12761322 TI - SIR 2003 film panel case 2: tumor emboli to the common iliac arteries. PMID- 12761323 TI - SIR 2003 film panel case 3: hemorrhagic hereditary telangectasia with median arcuate ligament compression. PMID- 12761324 TI - SIR 2003 film panel case 4: neurofibromatosis. PMID- 12761325 TI - Double wire technique for intracranial stent navigation. PMID- 12761326 TI - Acid tests of N-methyl-D-aspartate receptor gating basics. PMID- 12761327 TI - The evolutionarily triumphant G-protein-coupled receptor. PMID- 12761328 TI - Nitric oxide signaling in brain: potentiating the gain with YC-1. PMID- 12761329 TI - Cyclic nucleotide phosphodiesterase 5 and sildenafil: promises realized. PMID- 12761330 TI - Molecular determinants of proton-sensitive N-methyl-D-aspartate receptor gating. AB - Extracellular protons inhibit N-methyl-D-aspartate (NMDA) receptors with an IC50 value in the physiological pH range. To identify the molecular determinants of proton sensitivity, we used scanning mutagenesis of the NR1 subunit to search for residues that control proton inhibition of NMDA receptors. Homology modeling of the extracellular domains suggested that residues at which mutations perturbed pH sensitivity were localized in discrete regions. The majority of mutations that strongly affected proton sensitivity were clustered in the extracellular end of the second transmembrane domain (M3) and adjacent linker leading to the S2 portion of the glycine-binding domain of NR1. Mutations in NR2A confirmed that the analogous region controls the pH sensitivity of this subunit and also identified the linker region between the third transmembrane domain (M4) and the S2 portion of the NR2 glutamate binding domain as an additional determinant of proton sensitivity. One mutant receptor, NR1(A649C)/NR2A(A651T), showed a 145 fold reduction in the IC50 for protons (IC50, 17.3 microM corresponding to pH 4.9). The M3-S2 linker region has been suggested to control NMDA receptor gating, leading to the hypothesis that the proton sensor and receptor gate may be structurally and functionally integrated. PMID- 12761331 TI - 3,4-methylenedioxymethamphetamine (MDMA, "Ecstasy") induces fenfluramine-like proliferative actions on human cardiac valvular interstitial cells in vitro. AB - Recent findings have implicated the 5-hydroxytryptamine 2B (5-HT2B) serotonin receptor in mediating the heart valve fibroplasia [valvular heart disease (VHD)] and primary pulmonary hypertension observed in patients taking the now-banned appetite suppressant fenfluramine (Pondimin, Redux). Via large-scale, random screening of a portion of the receptorome, we have discovered that the amphetamine derivative 3,4-methylenedioxymethamphetamine (MDMA, "Ecstasy") and its N-demethylated metabolite 3,4-methylenedioxyamphetamine (MDA) each preferentially bind to and activate human recombinant 5-HT2B receptors. We also demonstrate that MDMA and MDA, like fenfluramine and its N-deethylated metabolite norfenfluramine, elicit prolonged mitogenic responses in human valvular interstitial cells via activation of 5-HT2B receptors. We also report that pergolide and dihydroergotamine, two drugs recently demonstrated to induce VHD in humans, potently activate 5-HT2B receptors, thus validating this assay system for its ability to predict medications that might induce VHD. Our discovery that MDMA and a major metabolite, MDA, induce prolonged mitogenic responses in vitro similar to those induced by fenfluramine and norfenfluramine in vivo (i.e., valvular interstitial cell fibroplasia) predict that long-term MDMA use could lead to the development of fenfluramine-like VHD. Because of the widespread abuse of MDMA, these findings have major public health implications. These findings also underscore the necessity of screening current and future drugs at h5-HT2B receptors for agonist actions before their use in humans. PMID- 12761332 TI - Structural evaluation of the agonistic action of a vitamin D analog with two side chains binding to the nuclear vitamin D receptor. AB - The vitamin D receptor (VDR) is one of the endocrine members of the nuclear receptor superfamily and has a characteristic high affinity for its natural ligand 1alpha,25-dihydroxyvitamin D3 [1alpha,25(OH)2D3]. From a mechanistic point of view, the most interesting analog of 1alpha,25(OH)2D3 is the one that carries two side chains, referred to as Gemini. In this study, molecular dynamics (MD) simulations of the Gemini-VDR complex were performed that demonstrated that the binding of a ligand with a 25% increased volume does not disturb the overall structure of the ligand-binding domain (LBD). It was found that one of the two side chains takes exactly the same position as the single side chain of the natural ligand, which suggests that the molecular mechanism of the agonism of Gemini is identical to that of 1alpha,25(OH)2D3. VDR single and double point mutants at L227, A303, I313, and L397 and in vitro and ex vivo assessment of their agonistic action confirmed the predictions of the MD simulations. Moreover, it was found that the second side chain of Gemini can choose between two binding positions within the ligand-binding pocket of the VDR. These two newly identified "corners" were characterized most specifically by the amino acids pairs L227/A303 and I313/L397. Therefore, Gemini is an important model compound that allows further insight into the molecular actions of the VDR but is, in parallel, also a promising precursor for the design of even more potent 1alpha,25(OH)2D3 analogs. PMID- 12761333 TI - Hypertonicity inhibits lipopolysaccharide-induced nitric oxide synthase expression in smooth muscle cells by inhibiting nuclear factor kappaB. AB - The expression of inducible nitric-oxide synthase (iNOS) in vascular smooth muscle cells leads to prolonged vasorelaxation in vivo and contributes to the profound vasodilation induced by bacterial lipopolysaccharide (LPS) in septic shock. This induction of iNOS depends, in large part, on activation of nuclear factor (NF)-kappaB. Hypertonicity regulates the activity of NF-kappaB in different cell lines; as such, we propose that it should also regulate the expression of iNOS. Thus, the goal of this study was to determine whether hypertonicity regulates iNOS expression and function in smooth muscle cells and to elucidate the mechanism(s) underlying this process. Treatment of hamster ductus deferens (DDT1MF-2) cells and porcine aortic smooth muscle cells with either mannitol (50 mM) or NaCl (50 mM) reduced LPS-stimulated iNOS expression and nitric oxide release. Both of these agents also reduced the activation of NF kappaB induced by LPS, tumor necrosis factor-alpha and interleukin-1beta in smooth muscle cells. This inhibitory action was caused by suppression of IkappaB alpha phosphorylation, a prerequisite for ubiquitination and degradation of this protein, and showed additivity with N-benzoyloxycarbonyl (Z)-Leu-Leu-leucinal (MG 132), an inhibitor of proteasomal degradation of IkappaB-alpha. Furthermore, exposure to mannitol inhibited the activity of IkappaB kinase, an enzyme involved in phosphorylation of IkappaB-alpha. Mannitol was unable to affect the induction of iNOS produced by overexpression of RelA in DDT1MF-2 cells, suggesting that this agent does not have additional downstream inhibitory actions on this activated NF-kappaB subunit. Taken together, these data suggest that these hypertonic solutions may prove useful as anti-inflammatory agents, especially against conditions associated with increased NF-kappaB activity. PMID- 12761334 TI - Non-nuclear localized human NOSII enhances the bioactivation and toxicity of tirapazamine (SR4233) in vitro. AB - Tirapazamine (TPZ) is the lead member of a class of bioreductive drugs currently in phase II and III clinical trials. TPZ requires metabolic activation to give a cytotoxic free radical species, and this hypoxia-mediated process is carried out by a variety of cellular reductases, including NADPH cytochrome c (P450) reductase (P540R). Nitric-oxide synthase (NOS) is widely expressed in human tumors, and this enzyme consists of an oxidase and a reductase domain, the latter showing striking homology to P450R. Thus, in this article, we have investigated the role of one of the cytosolic isoforms of NOS [inducible NOS (NOSII)] in the bioactivation of this DNA-damaging antitumor agent. To achieve this, we have constitutively overexpressed NOSII in human breast tumor MDA231 cells by employing an optimized expression vector in which the strong human polypeptide chain elongation factor 1alpha promoter drives a bicistronic message encoding the genes for human NOSII and the puromycin-resistant gene (pac). Subcellular localization of NOSII in the stably transfected clones was determined after differential centrifugation and showed that NOSII catalytic activity was exclusively cytosolic as determined by conventional activity assay. This was confirmed by immunostaining followed by fluorescent microscopy studies. The increase in NOSII activity in a series of transfected clones was associated with an increase in TPZ metabolism and toxicity under hypoxic conditions. There was no similar increase in aerobic toxicity. These findings are of significance for two reasons. First, cellular NOSII activity, similar to that seen in human breast cancer, could contribute to TPZ toxicity; second, this will be a result of NOS derived/cytosol-associated TPZ radicals. PMID- 12761335 TI - The G-protein-coupled receptors in the human genome form five main families. Phylogenetic analysis, paralogon groups, and fingerprints. AB - The superfamily of G-protein-coupled receptors (GPCRs) is very diverse in structure and function and its members are among the most pursued targets for drug development. We identified more than 800 human GPCR sequences and simultaneously analyzed 342 unique functional nonolfactory human GPCR sequences with phylogenetic analyses. Our results show, with high bootstrap support, five main families, named glutamate, rhodopsin, adhesion, frizzled/taste2, and secretin, forming the GRAFS classification system. The rhodopsin family is the largest and forms four main groups with 13 sub-branches. Positions of the GPCRs in chromosomal paralogons regions indicate the importance of tetraploidizations or local gene duplication events for their creation. We also searched for "fingerprint" motifs using Hidden Markov Models delineating the putative inter relationship of the GRAFS families. We show several common structural features indicating that the human GPCRs in the GRAFS families share a common ancestor. This study represents the first overall map of the GPCRs in a single mammalian genome. Our novel approach of analyzing such large and diverse sequence sets may be useful for studies on GPCRs in other genomes and divergent protein families. PMID- 12761336 TI - Diazonamide A and a synthetic structural analog: disruptive effects on mitosis and cellular microtubules and analysis of their interactions with tubulin. AB - The marine ascidian Diazona angulata was the source organism for the complex cytotoxic peptide diazonamide A. The molecular structure of this peptide was recently revised after synthesis of a biologically active analog of diazonamide A in which a single nitrogen atom was replaced by an oxygen atom. Diazonamide A causes cells to arrest in mitosis, and, after exposure to the drug, treated cells lose both interphase and spindle microtubules. Both diazonamide A and the oxygen analog are potent inhibitors of microtubule assembly, equivalent in activity to dolastatin 10 and therefore far more potent than dolastatin 15. This inhibition of microtubule assembly is accompanied by potent inhibition of tubulin-dependent GTP hydrolysis, also comparable with the effects observed with dolastatin 10. However, the remaining biochemical properties of diazonamide A and its analog differ markedly from those of dolastatin 10 and closely resemble the properties of dolastatin 15. Neither diazonamide A nor the analog inhibited the binding of [3H]vinblastine, [3H]dolastatin 10, or [8-14C]GTP to tubulin. Nor were they able to stabilize the colchicine binding activity of tubulin. These observations indicate either that diazonamide A and the analog have a unique binding site on tubulin differing from the vinca alkaloid and dolastatin 10 binding sites, or that diazonamide A and the analog bind weakly to unpolymerized tubulin but strongly to microtubule ends. If the latter is correct, diazonamide A and its oxygen analog should have uniquely potent inhibitory effects on the dynamic properties of microtubules. PMID- 12761337 TI - Decrease in survival threshold of quiescent colon carcinoma cells in the presence of a small molecule integrin antagonist. AB - The role of adhesion molecules, such as alphav integrins, in the control of the survival of quiescent tumor cells is unclear. We used S 34961, a novel small molecule alphav integrin antagonist, to investigate the role of integrin signaling in the survival of populations of quiescent human HT-29 and HCT 116 colon carcinoma cells. S 34961 at 1 microM induced detachment, but cells retained viability, existing as clusters. Nonligated beta-integrins may recruit and activate caspase-8 [J Cell Biol 155:459-470, 2001]. However, congruent with the absence of apoptosis, no activation of caspase-8 in these cells was detected after incubation with S 34961. A rapid (2 h) change in conformation of the N terminus of proapoptotic Bak was observed before detachment, together with a decrease in phosphorylation of focal adhesion kinase (2 h) and subsequent (8 h) decreases in phosphorylation of extracellular signal-regulated kinase-1/2 and Akt. Together, these results suggested that although treatment with S 34961 has no effect on survival per se, it may reduce the survival threshold of the tumor cells, with Bak in an activated state. Indeed, concomitant incubation of S 34961 with 10 microM U-0126 (a mitogen-activated protein kinase kinase inhibitor) was found to lead to apoptosis (at 24 h), whereas U-0126 alone had no effect. Together, these observations could guide the use of combination therapy with integrin antagonists in the clinic. PMID- 12761338 TI - Inhibition of human insulin gene transcription by the immunosuppressive drugs cyclosporin A and tacrolimus in primary, mature islets of transgenic mice. AB - Cyclosporin A and tacrolimus are clinically important immunosuppressive drugs. They share a diabetogenic action as one of their most serious adverse effects. The underlying mechanism is unknown. Previous studies have shown that tacrolimus can inhibit insulin gene transcription at high concentrations in tumor cell lines. To study insulin gene transcription in normal, mature pancreatic islet cells, we used a novel approach in the present study. Transgenic mice that carry a human insulin promoter-reporter gene were generated. The human insulin promoter directed transcription in pancreatic islets and conferred a normal, physiological glucose response to reporter gene expression in isolated islets. After stimulation with glucose, human insulin promoter-mediated gene expression was inhibited in normal, mature islet cells by both tacrolimus and cyclosporin A to a large extent (approximately 70%) and with high potency at concentrations that are known to inhibit calcineurin phosphatase activity (IC50 values of 1 and 35 nM, respectively). Furthermore, glucose stimulated calcineurin phosphatase activity in mouse pancreatic islets, further supporting the view that calcineurin phosphatase activity is an essential part of glucose signaling to the human insulin gene. The high potency of cyclosporin A and tacrolimus in normal islets suggests that inhibition of insulin gene transcription by cyclosporin A and tacrolimus is clinically important and is one mechanism of the diabetogenic effect of these immunosuppressive drugs. PMID- 12761339 TI - Thyroid hormone induction of the adrenoleukodystrophy-related gene (ABCD2). AB - X-linked adrenoleukodystrophy (X-ALD) is a demyelinating disorder associated with impaired very-long-chain fatty-acid (VLCFA) beta-oxidation caused by mutations in the ABCD1 (ALD) gene that encodes a peroxisomal membrane ABC transporter. ABCD2 (ALDR) displays partial functional redundancy because when overexpressed, it is able to correct the X-ALD biochemical phenotype. The ABCD2 promoter contains a putative thyroid hormone-response element conserved in rodents and humans. In this report, we demonstrate that the element is capable of binding retinoid X receptor and 3,5,3'-tri-iodothyronine (T3) receptor (TRbeta) as a heterodimer and mediating T3 responsiveness of ABCD2 in its promoter context. After a T3 treatment, an induction of the ABCD2 gene was observed in the liver of normal rats but not that of TRbeta-/- mice. ABCD2 was not induced in the brain of the T3 treated rats. However, we report for the first time that induction of the ABCD2 redundant gene is feasible in myelin-producing cells (differentiated CG4 oligodendrocytes). The induction was specific for this cell type because it did not occur in astrocytes. Furthermore, we observed T3 induction of ABCD2 in human and mouse ABCD1-deficient fibroblasts, which was correlated with normalization of the VLCFA beta-oxidation. Finally, ABCD3 (PMP70), a close homolog of ABCD2, was also induced by T3 in the liver of control rats, but not that of TRbeta-/- mice, and in CG4 oligodendrocytes. PMID- 12761340 TI - 2-aminoethoxydiphenyl borane activates a novel calcium-permeable cation channel. AB - The membrane-permeable, noncompetitive inositol 1,4,5-trisphosphate (IP3) receptor inhibitor 2-aminoethoxydiphenyl borane (2-APB) has been widely used to probe for IP3-receptor involvement in calcium signaling pathways. However, a number of recent studies in different cell types revealed other sites of action of 2-APB. In this study, we examined the influence of 2-APB on capacitative calcium entry and intracellular Ca2+ concentrations in rat basophilic leukemia (RBL-2H3 m1) cells. 2-APB was found to inhibit capacitative calcium entry, but at concentrations greater than 50 microM, a new effect of 2-APB was observed. When capacitative calcium entry was blocked with Gd3+, 2-APB caused an increase in cytoplasmic Ca2+. This increase in intracellular Ca2+ was not caused by altered buffering of cytoplasmic Ca2+ and was not caused by or in any way affected by the depletion of intracellular Ca2+ stores. Associated with the increase in intracellular Ca2+, in the presence of 2 mM Ca2+, 2-APB activated single channels in the plasma membrane with a conductance of approximately 50 pS. These channels seem to be nonselective cation channels; monovalent cations are the major carriers of current, but finite permeability to Ca2+ leads to a significant intracellular Ca2+ signal. Experiments with excised patches indicate that 2-APB activates these channels from the outer aspect of the cell membrane. This effect of 2-APB further illustrates the complex actions of this compound and reveals the presence in RBL-2H3 m1 cells of a novel, ligand-gated calcium-permeable channel. PMID- 12761341 TI - Agonist actions of "beta-blockers" provide evidence for two agonist activation sites or conformations of the human beta1-adrenoceptor. AB - Previous work with 4-[3-[(1,1-dimethylethyl)amino]2-hydroxypropoxy]-1,3-dihydro 2H-benzimidazol-2-one (CGP 12177) has led to the suggestion that there are two different agonist conformations of the human beta1-adrenoceptor: 1) where classic agonists (catecholamines) and beta-antagonists act, and 2) where CGP 12177 is an agonist and relatively resistant to inhibition by beta-adrenoceptor antagonists. In the present study, we have used studies of cAMP response element-regulated gene transcription to confirm the presence of these two beta1-adrenoceptor sites/conformations and to provide strong evidence that a range of clinically used beta-adrenoceptor blockers (beta-blockers) exhibit differential agonists and/or antagonist actions at the two sites. [2-(3-Carbamoyl-4-hydroxyphenoxy) ethylamino]-3-[4-(1-methyl-4-trifluormethyl-2-imidazolyl)-phenoxy]-2 propanolmethanesulphonate (CGP 20712A) and atenolol act as classic antagonists at the catecholamine binding site but have much lower affinity for the secondary CGP 12177 site. CGP 12177 and carvedilol are potent antagonists at the catecholamine site but mediate substantial agonist actions on gene transcription via the secondary antagonist-resistant site at higher concentrations. Agonist effects of beta-blockers are not, however, confined to this secondary site, and we show that some (particularly acebutolol and labetolol) act primarily via the catecholamine site, whereas others (pindolol and alprenolol) can stimulate both. The different responses to beta-blockers seen in the clinic may therefore be caused in part by these beta-blocker agonist responses and the differential activation of the two sites or conformations. PMID- 12761342 TI - Enhancement of long-term potentiation by a potent nitric oxide-guanylyl cyclase activator, 3-(5-hydroxymethyl-2-furyl)-1-benzyl-indazole. AB - Nitric oxide (NO) is known to affect synaptic plasticity in various regions of the brain via the cGMP-cGMP-dependent protein kinase (PKG) pathway. We found that a novel compound 3-(5-hydroxymethyl-2-furyl)-1-benzyl-indazole (YC-1), a drug known to modulate the response of soluble guanylyl cyclase to NO, greatly potentiates long-term potentiation (LTP). This compound markedly enhanced the induction of LTP in rat hippocampal and amygdala slices by weak tetanic stimulation. The potentiation of LTP by YC-1 was greatly reduced by NO synthase inhibitor Ng-nitro-l-arginine-methylester, guanylyl cyclase inhibitor 1 H-[1,2,4] oxadiazolo(4,3-a)-quinoxalin-1-one, and PKG inhibitor (9S,10R,12R) 2,3,9,10,11,12, hexahydro-10-methoxy-2,9-dimethyl-1-ox0-9.12-epoxy-1H diindolo[1,2,3-fg:3',2',1'-kl]pyrrolo[3,4-I][1,6]benzodiazocine-10-carboxylic acid methyl ester (KT5823). In addition, mitogen-activated protein kinase kinase inhibitor 2'-amino-3'-methoxyflavone (PD98059) also markedly inhibited LTP potentiating action of YC-1. Intracellular increase of Ca2+ concentration derived from N-methyl-d-aspartate and glutamate metabotropic receptors contributes to the potentiating action of YC-1. Concurrent perfusion of YC-1 and NO donor sodium nitroprusside for a short time period resulted in the induction of LTP by stimuli at a frequency as low as 0.02 Hz. Incubation of unstimulated hippocampal slices with YC-1 plus nitroprusside increased the immunofluorescence of phospho extracellular signal-regulated kinase (ERK) and phospho-cAMP response element binding protein (CREB). Furthermore, the Western blot shows that the phosphorylation of ERKs 1 and 2 and CREB of unstimulated hippocampal slices was increased by YC-1 plus nitroprusside, which was inhibited by KT5823. The NO-cGMP PKG-ERK signaling pathway thus plays important role in the potentiation of LTP by YC-1. PMID- 12761343 TI - Developmental expression of heteromeric nicotinic receptor subtypes in chick retina. AB - Acting through nicotinic acetylcholine receptors (nAChRs), acetylcholine plays an important role in retinal development and the formation of retinal connections to target tissues, but very little is known about the nAChR subtypes expressed in vertebrate retina during neuronal development. We used immunoprecipitation and [3H]epibatidine binding to study the expression of chick retina alpha bungarotoxin-insensitive heteromeric nAChRs during development and adulthood, and found that it is strictly developmentally regulated, reaching a peak on postnatal day 1. The increase in [3H]epibatidine receptors is caused mainly by an increase in the receptors containing the alpha2, alpha6, beta3, and beta4 subunits. The contribution of beta subunits to [3H]epibatidine receptors significantly changes during development: the beta2 subunit is contained in the majority (84%) of receptors on embryonic day (E) 7 but in only 32% on postnatal day (P) 1, whereas the beta4-containing receptors increase from 22% to 78% during the same period. Using a sequential immunodepletion procedure, we purified the beta2- and beta4 containing subtypes and found that they coassemble with alpha4 and/or alpha3 on E11, and also with the alpha2, alpha6, and beta3 on P1. After the immunodepletion of alpha6-containing receptors, the beta2- and beta4-containing receptors have a very similar pharmacological profile on P1. Parallel immunoprecipitation experiments in other brain areas showed that the developmentally regulated receptors in optic lobe are those containing the alpha2, alpha5, and beta2 subunits and those containing the alpha4 and beta2 subunits, whereas the receptors in forebrain-cerebellum contain the alpha4 and beta2 subunits with or without the alpha5 subunit. These results indicate that there is an increase in receptor heterogeneity and complexity in chick retina during development that is also maintained in adulthood. PMID- 12761344 TI - Developmental changes in beta2-adrenergic receptor signaling in ventricular myocytes: the role of Gi proteins and caveolae microdomains. AB - Cardiomyocyte beta2-adrenergic receptors (beta-ARs) provide a source of inotropic support and influence the evolution of heart failure. Recent studies identify distinct mechanisms for beta2-AR actions in neonatal and adult rat cardiomyocytes. This study examines whether ontogenic changes in cardiac beta2-AR actions can be attributed to altered Gi expression or changes in the spatial organization of the beta2-AR complex in membrane subdomains (caveolae). We show that beta2-ARs increase cAMP, calcium, and contractile amplitude in a pertussis toxin (PTX)-insensitive manner in neonatal cardiomyocytes. This is not caused by lack of Gi; Galphai expression is higher in neonatal cardiomyocytes than in those of adult rats. beta2-ARs provide inotropic support without detectably increasing cAMP, in adult cardiomyocytes. This cannot be attributed to dual coupling of beta2-ARs to Gs and Gi, because beta2-ARs do not promote cAMP accumulation in PTX pretreated adult cardiomyocytes. Spatial segregation of beta2-ARs, Galphas/Galphai, and adenylyl cyclase to distinct membrane subdomains also is not a factor, because all of these proteins copurify in caveolin-3-enriched vesicles isolated from adult cardiomyocytes. However, these studies demonstrate that enzyme-based protocols routinely used to isolate ventricular cardiomyocytes lead to proteolysis of beta-ARs. The functional consequences of this limited beta-AR proteolysis is uncertain, because truncated beta1-ARs promote cAMP accumulation and truncated beta2-ARs provide inotropic support in adult cardiomyocytes. Collectively, these studies indicate that components of the beta2-AR signaling complex compartmentalize to restricted membrane subdomains in adult rat cardiomyocytes. Neither compartmentalization nor changes in Gi expression fully explain the ontogenic changes in beta2-AR responsiveness in the rat ventricle. PMID- 12761345 TI - Thiol-modifying phenylarsine oxide inhibits guanine nucleotide binding of Rho but not of Rac GTPases. AB - Phenylarsine oxide (PAO) is a phosphotyrosine phosphatase inhibitor that cross links vicinal thiol groups, thereby inactivating phosphatases possessing XCysXXCysX motifs. The RhoA-GTPase, but not the Rac1-GTPase, also possesses vicinal cysteines within the guanine nucleotide-binding region (aa 13-20) and the phosphohydrolase activity site. Treatment of Caco-2 cells with PAO showed a dose dependent reorganization of the actin cytoskeleton, indicating involvement of Rho GTPases. As tested by pull-down experiments, RhoA, but not Rac1, from cell lysates was inactivated by PAO in a concentration-dependent manner. Modification of RhoA by PAO resulted in altered mobility on SDS-polyacrylamide gel electrophoresis, and PAO-modified RhoA was no longer substrate for C3-catalyzed ADP-ribosylation. Furthermore, RhoA treated with PAO, but not Rac1 treated with PAO, lost its property to bind to guanine nucleotides. Matrix-assisted laser desorption ionization-mass analysis of PAO-modified RhoA showed a mass shift according to an adduction of a single PAO molecule per molecule RhoA. Further analysis of Glu-C-generated RhoA peptides confirmed binding of PAO to a peptide harboring the guanine nucleotide binding region. Thus, PAO does not exclusively inhibit phosphotyrosine phosphatases but also inactivates RhoA by alteration of nucleotide binding. PMID- 12761346 TI - Characterization of a Ca2+ response to both UTP and ATP at human P2Y11 receptors: evidence for agonist-specific signaling. AB - Previous reports on heterologously-expressed human P2Y11 receptors have indicated that ATP, but not UTP, is an agonist stimulating both phosphoinositidase C and adenylyl cyclase. Consistent with these findings, we report that in 1321N1 cells expressing human P2Y11 receptors, UTP stimulation did not lead to accumulation of inositol(poly)phosphates under conditions in which ATP gave a robust, concentration-dependent effect. Unexpectedly, however, both UTP and ATP stimulated increases in cytosolic Ca2+ concentration ([Ca2+]c), with both nucleotides achieving similar EC50 and maximal responses. The responses to maximally effective concentrations of ATP and UTP were not additive. The [Ca2+]c increase in response to UTP was less dependent on extracellular Ca2+ than was the response to ATP. AR-C67085 (2-propylthio-beta,gamma-difluoromethylene-d-ATP, a P2Y11-selective agonist), adenosine 5'-O-(3-thiotriphosphate), and benzoyl ATP were all full agonists with potencies similar to those of ATP and UTP. In desensitization experiments, exposure to ATP resulted in loss of the UTP response; this response was more sensitive to desensitization than that of ATP. Pertussis toxin pretreatment attenuated the response to UTP but left the ATP response unaffected. The presence of 2-aminoethyl diphenylborate differentially affected the responses of ATP and UTP. No mRNA transcripts for P2Y2 or P2Y4 were detectable in the P2Y11-expressing cells. We conclude that UTP is a Ca2+ mobilizing agonist at P2Y11 receptors and that ATP and UTP acting at the same receptor recruit distinct signaling pathways. This example of agonist-specific signaling is discussed in terms of agonist trafficking and differential signal strength. PMID- 12761347 TI - [3H]sildenafil binding to phosphodiesterase-5 is specific, kinetically heterogeneous, and stimulated by cGMP. AB - Sildenafil (Viagra) potentiates penile erection by acting as a nonhydrolyzable analog of cGMP and competing with this nucleotide for catalysis by phosphodiesterase-5 (PDE5), but the characteristics of direct binding of radiolabeled sildenafil to PDE5 have not been determined. [3H]Sildenafil binding to PDE5 was retained when filtered through nitrocellulose or glass-fiber membranes. Binding was inhibited by excess sildenafil, 2-(2-methylpyridin-4 yl)methyl-4-(3,4,5-trimethoxyphenyl)-8-(pyrimidin-2-yl)methoxy-1,2-dihydro-1-oxo 2,7-naphthyridine-3-carboxylic acid methyl ester hydrochloride (T-0156), 3 isobutyl-1-methylxanthine, EDTA, or cGMP, but not by cAMP or 5'-GMP. PDE5 was the only [3H]sildenafil binding protein detected in human lung extract. Using purified recombinant PDE5, [3H]sildenafil exchange dissociation yielded two components with t1/2 values of 1 and 14 min and corresponding calculated KD values of 12 and 0.83 nM, respectively. This implied the existence of two conformers of the PDE5 catalytic site. [3H]Sildenafil binding isotherm of PDE5 indicated KD was 8.3 to 13.3 nM, and low cGMP decreased the KD to 4.8 nM but only slightly increased Bmax to a maximum of 0.61 mol/mol-subunit. Results suggest that these effects occur via cGMP binding to the allosteric cGMP binding sites of PDE5. Results imply that by inhibiting PDE5 and thereby increasing cGMP, sildenafil accentuates its own binding affinity for PDE5, which further elevates cGMP. The data also indicate that after physiological elevation, cGMP may directly stimulate the catalytic site by binding to the allosteric cGMP-binding sites of PDE5, thus causing negative feedback on this pathway. PMID- 12761348 TI - Aryl hydrocarbon receptor gene silencing with small inhibitory RNA differentially modulates Ah-responsiveness in MCF-7 and HepG2 cancer cells. AB - Sequence-specific small interfering RNA (siRNA) duplexes can be used for gene silencing in mammalian cells and as mechanistic probes for determining gene function. Transfection of siRNAs for the aryl hydrocarbon receptor (AhR) and AhR nuclear translocator (ARNT) mRNAs in MCF-7 breast cancer cells resulted in a 60 to 80% decrease in levels of AhR and ARNT proteins in whole-cell extracts and decreased binding of nuclear extracts to 32P-labeled dioxin-responsive element. siRNA for the AhR also decreased 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) induced CYP1A1 protein, CYP1A1-dependent activity, and luciferase activity in cells transfected with an Ah-responsive construct. 17beta-estradiol (E2) induces proliferation of MCF-7 cells through enhanced G0/G1 --> S phase progression, and this response is inhibited in cells cotreated with E2 plus TCDD. The effects of TCDD on E2-induced cell-cycle progress were partially blocked in MCF-7 cells transfected with siRNA for AhR. The results also indicated that siRNA-dependent decreases in AhR protein in MCF-7 cells were accompanied by increased G0/G1 --> S phase progression, suggesting a growth-inhibitory role for the "endogenous" AhR. Surprisingly, TCDD alone induced G0/G1 --> S phase progression and exhibited estrogenic activity in MCF-7 cells transfected with siRNA for the AhR. In contrast, degradation of the AhR in HepG2 liver cancer cells resulted in decreased G0/G1 --> S phase progression, and this was accompanied by decreased expression of cyclin D1, cyclin E, cyclin-dependent kinase 2 (cdk2), and cdk4. In the absence of ligand, the AhR exhibits growth-inhibitory (MCF-7) and growth promoting (HepG2) activity that is cell context-dependent. PMID- 12761349 TI - DNA sequence specificity for topoisomerase II poisoning by the quinoxaline anticancer drugs XK469 and CQS. AB - The two known antineoplastic quinoxaline topoisomerase II poisons, XK469 (NSC 697887) and CQS (chloroquinoxaline sulfonamide, NSC 339004), were compared for DNA cleavage site specificity, using purified human topoisomerase IIalpha and human topoisomerase IIbeta. The DNA cleavage intensity pattern for topoisomerase IIalpha poisoning by CQS closely resembled that of VM-26, despite the lack of any apparent common pharmacophore. In contrast, the topoisomerase IIalpha DNA cleavage intensity patterns of XK469 and CQS were very different from one another despite the similar overall structures of the two drugs. This suggests that the differences in DNA site specificity of topoisomerase II poisoning by XK469 and CQS may be caused by differences in their geometry, side chains, or electronic structure. The topoisomerase IIbeta-mediated DNA cleavage sites of CQS and XK469 were also very different from one another, adding further support to this idea. Earlier work has demonstrated that a number of specific topoisomerase II poisons show very similar patterns of DNA cleavage with either topoisomerase IIalpha or topoisomerase IIbeta, suggesting that the topoisomerase II isozymes play only a minor role in choices of DNA cleavage sites. However, both of the quinoxaline topoisomerase II poisons in this study showed distinctly different and unique DNA cleavage intensity patterns with each topoisomerase II isozyme. This indicates that topoisomerase II isozymes can play a major role in DNA cleavage site selection for some classes of topoisomerase II poisons. PMID- 12761350 TI - Closure of gap junction channels by arylaminobenzoates. AB - We determined the effect of flufenamic acid (FFA) and related derivatives on gap junction channel currents, applying the dual whole-cell patch-clamp technique to pairs of N2A neuroblastoma cells transfected with various connexins. FFA reduced gap junction channel currents in a reversible and concentration-dependent manner. Half-maximal concentrations for FFA-induced reduction of junctional conductance in cell pairs coupled by different connexins were similar (20 to 60 microM), indicating that FFA does not greatly discriminate between connexin subtypes. Hill coefficients for blockade were approximately 3, indicating a high degree of cooperativity. Analogs of FFA also reduced junctional conductance with similar potencies, whereas other unrelated chloride channel blockers had no effect. Inhibition of gap junction channels by FFA (pKa approximately 3.8) was increased at low external pH, suggesting that the uncharged form of the drug is important for blockade. The effect of FFA did not seem to be mediated by direct binding of the drug to the pore of the gap junction channel. Internal application of high concentrations of FFA by addition to patch pipettes did not cause inhibition of channel currents. The magnitude of inhibition was neither voltage-dependent nor influenced by the nature of permeant ion. Single-channel recordings indicated that FFA reduced the channel-open probability without modifying the current amplitude and induced slow transitions between open and closed states. We propose that FFA inhibits gap junctions by inducing a conformational change in the protein upon binding to a site that is presumably located within the membrane. PMID- 12761351 TI - Point mutations at L1280 in Nav1.4 channel D3-S6 modulate binding affinity and stereoselectivity of bupivacaine enantiomers. AB - Local anesthetics (LAs) block voltage-gated sodium channels. Parts of the LA binding site are located in the pore-lining transmembrane segments 6 of domains 1, 3, and 4 (D1-S6, D3-S6, D4-S6). We suggested previously that residue N434 in D1-S6 interacts directly with bupivacaine enantiomers in inactivated channels because side-chain properties of different residues substituted at N434 correlated with changes in blocking potencies of bupivacaine enantiomers. Furthermore, mutation N434R exhibited significant stereoselectivity for block of inactivated channels that resulted from a selective decrease in block by S(-) bupivacaine. In the present study, we analyzed the role of residue L1280 in D3-S6 of the rat skeletal muscle Nav1.4 channel in interactions with the enantiomers of bupivacaine. We substituted native leucine at L1280 with amino acids of different physicochemical properties. Wild-type and mutant channels were expressed transiently in human embryonic kidney 293t cells and were investigated under whole-cell voltage clamp. Block of resting mutant channels by bupivacaine enantiomers revealed little difference compared with wild-type channels. Block of inactivated channels was increased in a mutation containing an aromatic group (L1280W) and decreased in mutations containing a positive charge (L1280K, L1280R). Surprisingly, mutants L1280E, L1280N, L1280Q, and L1280R exhibited significant stereoselectivity for block of inactivated channels. More surprisingly, stereoselectivity resulted from a selective decrease in block by R(+)-bupivacaine, in contrast to mutation N434R in D1-S6. We propose that in inactivated channels, residues L1280 in D3-S6 and N434 in D1-S6 interact directly with LAs and thereby face each other in the ion-conducting pore. PMID- 12761352 TI - Pharmacological and biophysical properties of the human P2X5 receptor. AB - We constructed a full-length human P2X5 purinoceptor cDNA by incorporating a sequence corresponding to exon 10, which is missing in cDNAs cloned previously from human tissues. We studied the functional properties by patch-clamp recording and fluorescence imaging after expression in human embryonic kidney 293 cells. ATP (1-100 microM; half-maximal current at 4 microM) elicited inward currents at 60 mV; these persisted during brief (2 s) applications but declined during longer applications. The peak current was dependent on the holding potential and showed little rectification; however, both the desensitization during the application and the decline in the current when ATP was washed out were slower at +30 mV than at -60 mV. 2',3'-O-(4-Benzoyl)-benzoyl-ATP and alphabeta-methylene-ATP mimicked the action of ATP (half-maximal concentrations 6 and 161 microM, respectively). The currents were inhibited by suramin, pyridoxal-5-phosphate-6-azo-2',4' disulfonic acid and Brilliant Blue G, with half-maximal inhibition at 3, 0.2, and 0.5 microM, respectively; 2',3'-O-(2',4',6'-trinitrophenol)-ATP (1 microM) was ineffective. Removing divalent cations did not significantly alter ATP concentration-response curves. Reversal potential measurements showed that the human P2X5 receptor was permeable to calcium (PCa/PNa = 1.5) and N-methyl-d glucamine (NMDG) (PNMDG/PNa = 0.4); it was also permeable to chloride (PCl/PNa = 0.5) but not gluconate (Pgluc/PNa = 0.01) ions. The permeability to NMDG developed as quickly as the channel opened, in contrast to the P2X7 receptor where the NMDG permeability develops over several seconds. Cells expressing human P2X5 receptors also rapidly accumulated the propidium dye YO-PRO-1 in response to ATP. PMID- 12761353 TI - Local anesthetic block of Kv channels: role of the S6 helix and the S5-S6 linker for bupivacaine action. AB - To gain insights in the molecular mechanisms of anesthesia, we analyzed the effects of bupivacaine on a series of voltage-gated K+ channels (Kv1.1, -1.2, 1.5, -2.1, -3.1, and -3.2) and various mutant channels derived from Kv2.1, using Xenopus laevis oocytes. Two phenomenologically different blocking effects were seen at room temperature: a time-dependent block of Kv1 and Kv3 channels (Kd between 110 and 240 microM), and a time-independent block on Kv2.1 (Kd = 220 microM). At 32 degrees C, however, Kv2.1 also showed a time-dependent block. Swapping the S6 helix between Kv1.2 and Kv2.1 introduced Kv1.2 features in Kv2.1. Critical residues were located in the N-terminal end of S6, positions 395 and 398. The triple substitution of residues 372, 373, and 374 in the S5-S6 linker decreased the bupivacaine affinity by 5-fold (Kd increased from 220 to 1170 microM). The results suggest that bupivacaine blocks Kv channels by an open-state dependent mechanism and that Kv2.1 deviates from the other channels in allowing a partial closure of the channel with bupivacaine bound. The results also suggest that the binding site is located in the internal vestibule and that residues in the descending P-loop and the upper part of S6 are critical for the binding, most likely by allosteric mechanisms. A simple mechanistic scenario that explains the observations is presented. Thermodynamic considerations suggest that the interaction between bupivacaine and the channels is hydrophobic. PMID- 12761354 TI - Extra- and intracellular sphingosylphosphorylcholine promote spontaneous transmitter release from frog motor nerve endings. AB - Similar to phosphatidylinositol bisphosphate, sphingomyelin breakdown generates several lipids, including sphingosylphosphorylcholine (SPC), that are putative signaling molecules. The present study was undertaken to evaluate the involvement of SPC in transmitter release process. Intracellular recordings were made from isolated frog sciatic-sartorius nerve-muscle preparations, and the effects of SPC on neurosecretion in the form of miniature endplate potentials (MEPPs) were assessed. Extracellular application of SPC mixture (D,L-SPC) at 1, 10, and 25 microM increased the MEPP frequency by 68, 96, and 127%, respectively. D-erythro SPC (dissolved in dimethyl sulfoxide but not coupled to bovine serum albumin), but not L-threo-SPC, was active extracellular; the former (at 10 microM) increased the MEPP frequency by 143%. D-erythro-SPC treatment did not significantly change the median amplitude or frequency-distribution of MEPPs. Intracellular delivery via liposomes, in which 10, 100, or 1000 microM SPC mixture was entrapped in liposomal aqueous phase, induced a concentration dependent increase in MEPP frequency of 45, 91, and 100%, respectively. D-erythro SPC and L-threo-SPC at the concentration of 100 microM increased the MEPP frequency by 117 and 67%, respectively, or 91 and 61%, respectively, when coupled to bovine serum albumin. Pretreatment with thapsigargin significantly reduced but did not abolish the effects of extracellular D-erythro-SPC (10 microM) or liposomes containing 100 microM D-erythro-SPC. Liposomes filled with 100 microM D myo-inositol 1,4,5-trisphosphate (IP3) enhanced the MEPP frequency to the same magnitude as 100 microM D-erythro-SPC entrapped in liposomes. However, administration of 100 microM D-erythro-SPC and IP3 entrapped in the same liposomes enhanced the MEPP frequency by 70%, which was less than that produced by these two compounds alone. The result provides the first electrophysiological evidence that SPC can modulate transmitter release by an extra- or intracellular action at the frog motor nerve ending. PMID- 12761355 TI - A key role for caspase-2 and caspase-3 in the apoptosis induced by 2-chloro-2' deoxy-adenosine (cladribine) and 2-chloro-adenosine in human astrocytoma cells. AB - Both the anticancer agent 2-chloro-2'-deoxy-adenosine (Cladribine) and its derivative 2-chloro-adenosine induce apoptosis of human astrocytoma cells (J Neurosci Res 60:388-400, 2000). In this study, we have analyzed the involvement of caspases in these effects. Both compounds produced a gradual and time dependent activation of "effector" caspase-3, which preceded the appearance of the nuclear signs of apoptosis, suggesting a temporal correlation between these two events. Moreover, the caspase inhibitor N-benzyloxycarbonyl-Val-Ala-dl-Asp fluoromethylketone (fmk) suppressed both caspase-3 activation and apoptosis induction. "Initiator" caspase-9 and caspase-8 were only marginally activated at later times in the apoptotic process. Accordingly, at concentrations that selectively inhibit these caspases, neither N-benzyloxycarbonyl-Leu-Glu-His-Asp fmk nor N-benzyloxycarbonyl-Ile-Glu-Thr-Asp-fmk could prevent adenosine analog induced cell death. To definitively rule out a role for the caspase-9/cytochrome c-dependent mitochondrial pathway of cell death, neither adenosine analog had any effect on mitochondrial membrane potential, which was instead markedly reduced by other apoptotic stimuli (e.g., deoxyribose, NaCN, and betulinic acid). Consistently, although the latter triggered translocation of mitochondrial cytochrome c to the cytoplasm, no cytosolic accumulation of cytochrome c was detected with adenosine analogs. Conversely, 1 to 7 h after addition of either adenosine analog (i.e., before the appearance of caspase-3 activation), caspase-2 activity was surprisingly and markedly increased. The selective caspase-2 inhibitor N-benzyloxy carbonyl-Val-Asp-Val-Ala-Asp-fmk significantly reduced both adenosine analogs-induced caspase-2 activation and the associated cell death. We conclude that adenosine analogs induce the apoptosis of human astrocytoma cells by activating an atypical apoptotic cascade involving caspase-2 as an initiator caspase, and effector caspase-3. Therefore, these compounds could be effectively used in the pharmacological manipulation of tumors characterized by resistance to cell death via either the mitochondrial or caspase-8/death receptor pathways. PMID- 12761356 TI - Hyperoxia causes inducible nitric oxide synthase-mediated cellular damage to the immature rat brain. AB - Relative hyperoxia is a condition frequently encountered in premature infants, either spontaneously or during treatment in the Neonatal Intensive Care Unit. The effects of high inspiratory oxygen concentrations on immature brain cells and their signaling cascades are largely unknown. The aim of the study was to investigate the effect of hyperoxia on the amount and topographic distribution of iNOS-expression (inducible nitric oxide synthase) in the immature rat brain, and to localize hyperoxia-induced formation of peroxynitrite as a potential marker of cellular damage to immature cerebral structures. Seven-day-old Wistar rat pups were exposed to >80% oxygen for 24 h and were then transcardially perfused. Following paraformaldehyde fixation, brains were paraffin-embedded and immunohistochemically stained for iNOS and nitrotyrosine. iNOS protein was quantified by Western blot; iNOS mRNA expression was studied by RT-PCR. Total brain iNOS mRNA was up-regulated, demonstrating a peak at 6 h following the onset of hyperoxia. Immunohistochemical staining was predominantly observed in microglial cells of hippocampus and frontal cortex with some iNOS reactivity in endothelial and perivascular cells. Nitrotyrosine staining was positive in apical dendrites of neurons in the frontal cortex. There was no positive staining for iNOS or nitrotyrosine in control animals. Hyperoxia causes iNOS mRNA and protein up-regulation in microglial cells of the immature rat brain. Positive neuronal nitrotyrosine staining indicates formation of peroxynitrite with potential deleterious effects for immature cellular structures in the neonatal brain. PMID- 12761357 TI - The Burkholderia cepacia complex and cytokine induction: an inflammatory tale. PMID- 12761358 TI - Burkholderia cepacia-induced IL-8 gene expression in an alveolar epithelial cell line: signaling through CD14 and mitogen-activated protein kinase. AB - Burkholderia cepacia is a prevalent pulmonary pathogen in patients with cystic fibrosis (CF). The lung pathology observed in patients with CF is postulated to be due to an overexpression of chemokines. This study investigated the induction of the neutrophil chemoattractant chemokine IL-8 and the signaling pathways activated by B. cepacia-infected human lung epithelial A549 (HLE) cells. Cells were infected with B. cepacia (genomovar III of the B. cepacia complex), and reverse transcriptase-PCR and ELISA for the cytokines were performed. B. cepacia (multiplicity of infection > or =4:1) induced HLE cells to significantly secrete IL-8 in a more potent manner than the predominant CF pathogen Pseudomonas aeruginosa (multiplicity of infection > or =64:1). IL-8 secretion by B. cepacia infected HLE cells was abrogated by the gene transcription inhibitor actinomycin D and the protein translation inhibitor cycloheximide, confirming that B. cepacia induced IL-8 secretion was mediated through de novo protein synthesis. Treatment of B. cepacia with proteinase K failed to down-regulate IL-8 secretion; furthermore, IL-8 secretion by B. cepacia-infected HLE cells was abrogated by > or =80% in the presence of anti-CD14 [specific lipopolysaccharide (LPS) receptor] antibody, thus suggesting that the IL-8-inducing component of B. cepacia was LPS and therefore dependent on CD14. The p38 mitogen-activated protein kinase (MAPK) inhibitor and the extracellular signal-regulated kinase MAPK inhibitor significantly abrogated IL-8 secretion by B. cepacia-infected HLE cells (SB203580, > or =80% inhibition; PD98059, > or =30% inhibition). In conclusion, B. cepacia-induced IL-8 secretion in A549 airway epithelial cells is more potent than P. aeruginosa; is mediated through LPS, which is CD14 dependent; and involves activation of the p38 and ERK MAPK pathways. PMID- 12761359 TI - Formation of advanced glycosylation end products and oxidative stress in young patients with type 1 diabetes. AB - Increased production of advanced glycosylation end products (AGEs) and augmented oxidative stress may contribute to vascular complications in diabetes. Little is known about the formation and accumulation of AGEs in young patients with type 1 diabetes. The aim of the present study was to investigate whether AGE production and oxidative stress are augmented in young patients with type 1 diabetes at early clinical stages of the disease. Urine samples of 38 patients with type 1 diabetes [mean age (+/-SD), 12.8 +/- 4.5 y; diabetes duration, 5.7 +/- 4.3 y; HbA1c, 8.0 +/- 1.6%; urinary albumin excretion, 12.6 +/- 14.4 mg/g creatinine (Cr)] and those of 60 age-matched healthy control subjects were assayed for AGEs, pentosidine and pyrraline, and markers of oxidative stress, 8-hydroxy-2' deoxyguanosine (8-OHdG) and acrolein-lysine. Of these four markers, urinary concentrations of pentosidine, 8-OHdG, and acrolein-lysine were significantly higher in the patients with diabetes than in the healthy control subjects. For the patient group, pentosidine correlated significantly with 8-OHdG and acrolein lysine, and pyrraline correlated significantly with acrolein-lysine. Urinary pentosidine, 8-OHdG, and acrolein-lysine but not pyrraline correlated significantly with urinary albumin excretion. Patients with microalbuminuria (> or =15 mg/g Cr) showed significantly higher levels of all four markers than did normoalbuminuric patients and control subjects. The present study indicates that accumulation of AGEs, whose formation is closely linked to oxidative stress, and resultant endothelial dysfunction may start early in the course of type 1 diabetes. This means that the risk of vascular complications may be present at an early age and that the best possible glycemic control should be emphasized from the diagnosis of diabetes. PMID- 12761361 TI - Effect of body temperature on the pattern of spontaneous breathing in extremely low birth weight infants supported by proportional assist ventilation. AB - The optimum body temperature for infants <1000 g is unknown. We investigated body temperature effects on spontaneous breathing using proportional assist ventilation (PAV), because this mode supports spontaneous breathing such that all breathing pattern variables remain controlled by the infant. Minute volume (MV), respiratory rate (RR), tidal volume (Vt), incidence and duration of respiratory pauses, arterial oxygen desaturations <85%, and arterial Pco2 levels will remain unaffected by targeting core body temperature to 36.1-36.5 degrees C (low normal range) versus 37.7-37.9 degrees C (upper normal). Twenty infants (mean +/- SD: birth weight, 696 +/- 155 g; gestational age, 25 +/- 1 wk; age, 5 +/- 3 d) who were supported by PAV were exposed to each target temperature range on 2 consecutive days in four 2-h intervals for a total of 8 h with the sequence of the temperature ranges randomized. Core body temperature was 36.5 +/- 0.2 degrees C and 37.9 +/- 0.2 degrees C in the two conditions. MV was 291 and 314 mL. min-1. kg-1, respectively (7% difference; p < 0.001) as a result of a difference in RR (8%; p < 0.001). The infants maintained their blood CO2 levels and Vt (5.25 +/- 0.6 versus 5.19 +/- 0.6 mL/kg). Incidence and duration of respiratory pauses were not different between conditions. Extremely immature infants who are supported by PAV modify their spontaneous breathing in response to changes in thermal environment such that Pco2 levels are appropriately maintained early in postnatal life. This response pattern occurred consistently and is currently of uncertain clinical significance. PMID- 12761360 TI - Analysis of circulating T gamma/delta lymphocytes and CD16/56 cell populations in children and adolescents with Graves' disease. AB - The aim of the present study was to determine the proportion of gamma/delta T lymphocytes and CD16/CD56 (CD3- and CD3+) cells in the peripheral blood of children and adolescents with Graves' disease (GD; n = 27; mean age, 15.5 +/- 5.1 y) and nontoxic nodular goiter (NTNG; n = 25; mean age, 15.2 +/- 5.7 y), in comparison with sex- and age-matched healthy control subjects (n = 25; mean age, 15.9 +/- 2.4 y). In addition, in patients with GD, we investigated the effect of methimazole therapy on the proportion of these cells. We also looked for associations among the parameters investigated. The percentages of gamma/delta TCR+CD3+ lymphocytes and CD3+, CD16/56+CD3+, and natural killer (NK) cells were analyzed by the three-color flow cytometry using a Coulter EPICS XL cytometer. In patients with untreated GD, we observed a significant decrease in gamma/delta T (CD3+) (p < 0.002), CD16/56(CD3+) (p < 0.001), and NK (p < 0.001) cells in comparison with the healthy control subjects. After 2-6 mo of methimazole therapy, the percentages of gamma/delta TCR+CD3+ and CD16/56(CD3+) cells in peripheral blood of hyperthyroid patients returned to the normal values, whereas the percentages of NK cells normalized after 18-24 mo of therapy. These abnormalities were absent in children and adolescents with NTNG. Furthermore, there was no difference in the percentage of CD3+ lymphocytes in all of the groups. In the patients with untreated GD, we found a negative correlation between free thyroxine concentration in blood serum and the percentages of CD16/56 (CD3-) and gamma delta T cells (r = -0.5, p < 0.035; r = -0.4, p < 0.02). No such correlation was detected in patients with NTNG. We conclude that the abnormal distribution of CD16/CD56 (CD3- and CD3+) cells and gamma/delta T lymphocytes in the peripheral blood in children and adolescents with untreated GD suggests their role in the development of autoimmunity. PMID- 12761362 TI - IL-1 beta depresses respiration and anoxic survival via a prostaglandin-dependent pathway in neonatal rats. AB - IL-1 beta has been proposed to be an important mediator linking infection, apnea, and sudden infant death syndrome. We hypothesized that IL-1 beta acts in this capacity by depressing brainstem respiratory neurons via a prostaglandin dependent pathway. For studying the effects of IL-1 beta on respiration as well as the mechanism underlying its actions, 7-d-old rats received an initial injection (i.p.) of NaCl or a cyclooxygenase inhibitor (indomethacin, 10 mg/kg) followed by a second injection (i.p.) at 30 min of NaCl, recombinant rat IL-1 beta (10 microg/kg), or lipopolysaccharide (LPS; 100 microg/kg). Respiration during normoxia and in response to anoxia (100% N2) was examined at 60 min after the second injection using flow and barometric plethysmography. Animals given IL 1 beta breathed more slowly and died more often after anoxia. LPS also reduced the rats' ability to autoresuscitate and survive an anoxic challenge. Indomethacin prevented the depressive effects during normoxia and the adverse effects on survival. For investigating drug-induced changes in central respiratory activity, IL- 1 beta (1.0 or 1.25 ng/mL) and prostaglandin E2 (5 or 20 microg/L) was applied to the brainstem-spinal cord preparation of 0- to 4-d old rats. Whereas IL-1 beta exerted no effect on respiration measured at the C4 ventral root during a 60-min period, prostaglandin E2 reversibly inhibited respiratory activity. These findings suggest that IL-1 beta does not inhibit respiratory neurons directly but may depress breathing and hypoxic defense via a prostaglandin-mediated mechanism. PMID- 12761363 TI - Diet, obesity, and cardiovascular risk. PMID- 12761364 TI - A low-carbohydrate as compared with a low-fat diet in severe obesity. AB - BACKGROUND: The effects of a carbohydrate-restricted diet on weight loss and risk factors for atherosclerosis have been incompletely assessed. METHODS: We randomly assigned 132 severely obese subjects (including 77 blacks and 23 women) with a mean body-mass index of 43 and a high prevalence of diabetes (39 percent) or the metabolic syndrome (43 percent) to a carbohydrate-restricted (low-carbohydrate) diet or a calorie- and fat-restricted (low-fat) diet. RESULTS: Seventy-nine subjects completed the six-month study. An analysis including all subjects, with the last observation carried forward for those who dropped out, showed that subjects on the low-carbohydrate diet lost more weight than those on the low-fat diet (mean [+/-SD], -5.8+/-8.6 kg vs. -1.9+/-4.2 kg; P=0.002) and had greater decreases in triglyceride levels (mean, -20+/-43 percent vs. -4+/-31 percent; P=0.001), irrespective of the use or nonuse of hypoglycemic or lipid-lowering medications. Insulin sensitivity, measured only in subjects without diabetes, also improved more among subjects on the low-carbohydrate diet (6+/-9 percent vs. -3+/-8 percent, P=0.01). The amount of weight lost (P<0.001) and assignment to the low-carbohydrate diet (P=0.01) were independent predictors of improvement in triglyceride levels and insulin sensitivity. CONCLUSIONS: Severely obese subjects with a high prevalence of diabetes or the metabolic syndrome lost more weight during six months on a carbohydrate-restricted diet than on a calorie- and fat restricted diet, with a relative improvement in insulin sensitivity and triglyceride levels, even after adjustment for the amount of weight lost. This finding should be interpreted with caution, given the small magnitude of overall and between-group differences in weight loss in these markedly obese subjects and the short duration of the study. Future studies evaluating long-term cardiovascular outcomes are needed before a carbohydrate-restricted diet can be endorsed. PMID- 12761366 TI - Images in clinical medicine. Centrilobular emphysema with predominantly upper lobe involvement. PMID- 12761365 TI - A randomized trial of a low-carbohydrate diet for obesity. AB - BACKGROUND: Despite the popularity of the low-carbohydrate, high-protein, high fat (Atkins) diet, no randomized, controlled trials have evaluated its efficacy. METHODS: We conducted a one-year, multicenter, controlled trial involving 63 obese men and women who were randomly assigned to either a low-carbohydrate, high protein, high-fat diet or a low-calorie, high-carbohydrate, low-fat (conventional) diet. Professional contact was minimal to replicate the approach used by most dieters. RESULTS: Subjects on the low-carbohydrate diet had lost more weight than subjects on the conventional diet at 3 months (mean [+/-SD], 6.8+/-5.0 vs. -2.7+/-3.7 percent of body weight; P=0.001) and 6 months (-7.0+/ 6.5 vs. -3.2+/-5.6 percent of body weight, P=0.02), but the difference at 12 months was not significant (-4.4+/-6.7 vs. -2.5+/-6.3 percent of body weight, P=0.26). After three months, no significant differences were found between the groups in total or low-density lipoprotein cholesterol concentrations. The increase in high-density lipoprotein cholesterol concentrations and the decrease in triglyceride concentrations were greater among subjects on the low carbohydrate diet than among those on the conventional diet throughout most of the study. Both diets significantly decreased diastolic blood pressure and the insulin response to an oral glucose load. CONCLUSIONS: The low-carbohydrate diet produced a greater weight loss (absolute difference, approximately 4 percent) than did the conventional diet for the first six months, but the differences were not significant at one year. The low-carbohydrate diet was associated with a greater improvement in some risk factors for coronary heart disease. Adherence was poor and attrition was high in both groups. Longer and larger studies are required to determine the long-term safety and efficacy of low-carbohydrate, high protein, high-fat diets. PMID- 12761367 TI - Clinical practice. Restless legs syndrome. PMID- 12761368 TI - Mechanisms of actions of inhaled anesthetics. PMID- 12761370 TI - Interpreting incomplete data in studies of diet and weight loss. PMID- 12761369 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 16-2003. A 58-year-old woman with left-sided weakness and a right frontal brain mass. PMID- 12761371 TI - Expression of concern: Schiffl H, et Al. Daily hemodialysis and the outcome of acute renal failure. N Engl J Med 2002;346:305-10. PMID- 12761372 TI - Genetic medicine and obesity. PMID- 12761373 TI - The dilemma of Medicaid. PMID- 12761374 TI - Survivors of the acute respiratory distress syndrome. PMID- 12761375 TI - Signaling molecules in pulmonary hypertension. PMID- 12761376 TI - Mercury and the risk of myocardial infarction. PMID- 12761377 TI - Prevention of eclampsia. PMID- 12761378 TI - Immunogenicity of infliximab in Crohn's disease. PMID- 12761379 TI - Thoracic echinococcosis. PMID- 12761380 TI - Corticosteroid insufficiency in acutely ill patients. PMID- 12761381 TI - Case 27-2002: late-onset infantile neuronal ceroid lipofuscinosis. PMID- 12761382 TI - Ghrelin concentrations in morbidly obese patients after adjustable gastric banding. PMID- 12761384 TI - The shape of the Neandertal femur is primarily the consequence of a hyperpolar body form. AB - Neandertal femora are distinct from contemporaneous near-modern human femora. Traditionally, these contrasts in femoral shape have been explained as the result of the elevated activity levels and limited cultural abilities of Neandertals. More recently, however, researchers have realized that many of these femoral differences may be explained by the cold-adapted bodies of Neandertals vs. the warm-adapted bodies of near-modern humans. This study explicitly tests this proposed link between climate-induced body proportions and femoral shape by considering the entire hip as a unit by using geometric morphometric methods adapted to deal with articulated structures. Based on recent human patterns of variation, most contrasts in shape between the femora of Neandertals and near modern humans seem to be secondary consequences of differences in climate-induced body proportions. These results, considered in light of hip mechanics during growth, highlight the importance of developmental and functional integration in determining skeletal form. PMID- 12761383 TI - Hepatitis C virus glycoproteins mediate pH-dependent cell entry of pseudotyped retroviral particles. AB - HIV pseudotypes bearing native hepatitis C virus (HCV) glycoproteins (strain H and Con1) are infectious for the human hepatoma cell lines Huh-7 and PLC/PR5. Infectivity depends on coexpression of both E1 and E2 glycoproteins, is pH dependent, and can be neutralized by mAbs mapping to amino acids 412-447 within E2. Cell-surface expression of one or all of the candidate receptor molecules (CD81, low-density lipoprotein receptor, scavenger receptor class B type 1, and dendritic cell-specific intercellular adhesion molecule 3 grabbing nonintegrin) failed to confer permissivity to HIV-HCV pseudotype infection. However, HIV-HCV pseudotype infectivity was inhibited by a recombinant soluble form of CD81 and a mAb specific for CD81, suggesting that CD81 may be a component of a receptor complex. PMID- 12761385 TI - The role of alpha-, 3(10)-, and pi-helix in helix-->coil transitions. AB - The conformational equilibrium between 3(10)- and alpha-helical structure has been studied via high-resolution NMR spectroscopy by Millhauser and coworkers using the MW peptide Ac-AMAAKAWAAKA AAARA-NH2. Their 750-MHz nuclear Overhauser effect spectroscopy (NOESY) spectra were interpreted to reflect appreciable populations of 3(10)-helix throughout the peptide, with the greatest contribution at the N and C termini. The presence of simultaneous alphaN(i,i + 2) and alphaN(i,i + 4) NOE cross-peaks was proposed to represent conformational averaging between 3(10)- and alpha-helical structures. In this study, we describe 25-nsec molecular dynamics simulations of the MW peptide at 298 K, using both an 8 A and a 10 A force-shifted nonbonded cutoff. The ensemble averages of both simulations are in reasonable agreement with the experimental helical content from circular dichroism (CD), the (3)J(HNalpha) coupling constants, and the 57 observed NOEs. Analysis of the structures from both simulations revealed very little formation of contiguous i --> i + 3 hydrogen bonds (3(10)-helix); however, there was a large population of bifurcated i --> i + 3 and i --> i + 4 alpha helical hydrogen bonds. In addition, both simulations contained considerable populations of pi-helix (i --> i + 5 hydrogen bonds). Individual turns formed over residues 1-9, which we predict contribute to the intensities of the experimentally observed alphaN(i,i + 2) NOEs. Here we show how sampling of both folded and unfolded structures can provide a structural framework for deconvolution of the conformational contributions to experimental ensemble averages. PMID- 12761386 TI - Fishing new proteins in the twilight zone of genomes: the test case of outer membrane proteins in Escherichia coli K12, Escherichia coli O157:H7, and other Gram-negative bacteria. AB - We address the problem of clustering the whole protein content of genomes into three different categories-globular, all-alpha, and all-beta membrane proteins with the aim of fishing new membrane proteins in the pool of nonannotated proteins (twilight zone). The focus is then mainly on outer membrane proteins. This is performed by using an integrated suite of programs (Hunter) specifically developed for predicting the occurrence of signal peptides in proteins of Gram negative bacteria and the topography of all-alpha and all-beta membrane proteins. Hunter is tested on the well and partially annotated proteins (2160 and 760, respectively) of Escherichia coli K 12 scoring as high as 95.6% in the correct assignment of each chain to the category. Of the remaining 1253 nonannotated sequences, 1099 are predicted globular, 136 are all-alpha, and 18 are all-beta membrane proteins. In Escherichia coli 0157:H7 we filtered 1901 nonannotated proteins. Our analysis classifies 1564 globular chains, 327 inner membrane proteins, and 10 outer membrane proteins. With Hunter, new membrane proteins are added to the list of putative membrane proteins of Gram-negative bacteria. The content of outer membrane proteins per genome (nine are analyzed) ranges from 1.5% to 2.4%, and it is one order of magnitude lower than that of inner membrane proteins. The finding is particularly relevant when it is considered that this is the first large-scale analysis based on validated tools that can predict the content of outer membrane proteins in a genome and can allow cross-comparison of the same protein type between different species. PMID- 12761387 TI - Noncharged amino acid residues at the solvent-exposed positions in the middle and at the C terminus of the alpha-helix have the same helical propensity. AB - It was established previously that helical propensities of different amino acid residues in the middle of alpha-helix in peptides and in proteins are very similar. The statistical analysis of the protein helices from the known three dimensional structures shows no difference in the frequency of noncharged residues in the middle and at the C terminus. Yet, experimental studies show distinctive differences for the helical propensities of noncharged residues in the middle and in the C terminus in model peptides. Is this a general effect, and is it applicable to protein helices or is it specific to the model alanine-based peptides? To answer this question, the effects of substitutions at positions 28 (middle residue) and 32 (C2 position at the C terminus) of the alpha-helix of ubiquitin on the stability of this protein are measured by using differential scanning calorimetry. The two data sets produce similar values for intrinsic helix propensity, leading to a conclusion that noncharged amino acid residues at the solvent-exposed positions in the middle and at the C terminus of the alpha helix have the same helical propensity. This conclusion is further supported with an excellent correlation between the helix propensity scale obtained for the two positions in ubiquitin with the experimental helix propensity scale established previously and with the statistical distribution of the residues in protein helices. PMID- 12761388 TI - Reducing the computational complexity of protein folding via fragment folding and assembly. AB - Understanding, and ultimately predicting, how a 1-D protein chain reaches its native 3-D fold has been one of the most challenging problems during the last few decades. Data increasingly indicate that protein folding is a hierarchical process. Hence, the question arises as to whether we can use the hierarchical concept to reduce the practically intractable computational times. For such a scheme to work, the first step is to cut the protein sequence into fragments that form local minima on the polypeptide chain. The conformations of such fragments in solution are likely to be similar to those when the fragments are embedded in the native fold, although alternate conformations may be favored during the mutual stabilization in the combinatorial assembly process. Two elements are needed for such cutting: (1) a library of (clustered) fragments derived from known protein structures and (2) an assignment algorithm that selects optimal combinations to "cover" the protein sequence. The next two steps in hierarchical folding schemes, not addressed here, are the combinatorial assembly of the fragments and finally, optimization of the obtained conformations. Here, we address the first step in a hierarchical protein-folding scheme. The input is a target protein sequence and a library of fragments created by clustering building blocks that were generated by cutting all protein structures. The output is a set of cutout fragments. We briefly outline a graph theoretic algorithm that automatically assigns building blocks to the target sequence, and we describe a sample of the results we have obtained. PMID- 12761389 TI - An electronic effect on protein structure. AB - The well-known preference of the peptide bond for the trans conformation has been attributed to steric effects. Here, we show that a proline residue with an N formyl group (H(i-1)-C'(i-1)=O(i-1)), in which H(i-1) presents less steric hindrance than does O(i-1), likewise prefers a trans conformation. Thus, the preference of the peptide bond for the trans conformation cannot be explained by steric effects alone. Rather, an n --> pi* interaction between the oxygen of the peptide bond (O(i-1)), and the subsequent carbonyl carbon in the polypeptide chain (C'(i)) also contributes to this preference. The O(i-1) and C'(i) distance and O(i-1).C'(i)=O(i) angle are especially favorable for such an n --> pi* interaction in a polyproline II helix. We propose that this electronic effect provides substantial stabilization to this and other elements of protein structure. PMID- 12761390 TI - Structure of two fungal beta-1,4-galactanases: searching for the basis for temperature and pH optimum. AB - beta-1,4-Galactanases hydrolyze the galactan side chains that are part of the complex carbohydrate structure of the pectin. They are assigned to family 53 of the glycoside hydrolases and display significant variations in their pH and temperature optimum and stability. Two fungal beta-1,4-galactanases from Myceliophthora thermophila and Humicola insolens have been cloned and heterologously expressed, and the crystal structures of the gene products were determined. The structures are compared to the previously only known family 53 structure of the galactanase from Aspergillus aculeatus (AAGAL) showing approximately 56% identity. The M. thermophila and H. insolens galactanases are thermophilic enzymes and are most active at neutral to basic pH, whereas AAGAL is mesophilic and most active at acidic pH. The structure of the M. thermophila galactanase (MTGAL) was determined from crystals obtained with HEPES and TRIS buffers to 1.88 A and 2.14 A resolution, respectively. The structure of the H. insolens galactanase (HIGAL) was determined to 2.55 A resolution. The thermostability of MTGAL and HIGAL correlates with increase in the protein rigidity and electrostatic interactions, stabilization of the alpha-helices, and a tighter packing. An inspection of the active sites in the three enzymes identifies several amino acid substitutions that could explain the variation in pH optimum. Examination of the activity as a function of pH for the D182N mutant of AAGAL and the A90S/ H91D mutant of MTGAL showed that the difference in pH optimum between AAGAL and MTGAL is at least partially associated with differences in the nature of residues at positions 182, 90, and/or 91. PMID- 12761391 TI - Insights into dimerization and four-helix bundle formation found by dissection of the dimer interface of the GrpE protein from Escherichia coli. AB - The GrpE heat shock protein from Escherichia coli has a homodimeric structure. The dimer interface encompasses two long alpha-helices at the NH(2)-terminal end from each monomer (forming a "tail"), which lead into a small four-helix bundle from which each monomer contributes two short sequential alpha-helices in an antiparallel topological arrangement. We have created a number of different deletion mutants of GrpE that have portions of the dimer interface to investigate requirements for dimerization and to study four-helix bundle formation. Using chemical crosslinking and analytical ultracentrifugation techniques to probe for multimeric states, we find that a mutant containing only the long alpha-helical tail portion (GrpE1-88) is unable to form a dimer, most likely due to a decrease in alpha-helical content as determined by circular dichroism spectroscopy, thus one reason for a dimeric structure for the GrpE protein is to support the tail region. Mutants containing both of the short alpha-helices (GrpE1-138 and GrpE88 197) are able to form a dimer and presumably the four-helix bundle at the dimer interface. These two mutants have equilibrium constants for the monomer-dimer equilibrium that are very similar to the full-length protein suggesting that the tail region does not contribute significantly to the stability of the dimer. Interestingly, one mutant that contains just one of the short alpha-helices (GrpE1-112) exists as a tetrameric species, which presumably is forming a four helix bundle structure. A proposed model is discussed for this mutant and its relevance for factors influencing four-helix bundle formation. PMID- 12761392 TI - A wheat germ cell-free system is a novel way to screen protein folding and function. AB - For high-throughput protein structural analysis, it is indispensable to develop a reliable protein overexpression system. Although many protein overexpression systems, such as that involving Escherichia coli cells, have been developed, the number of overexpressed proteins showing the same biological activities as those of the native proteins is limited. A novel wheat germ cell-free protein synthesis system was developed recently, and most of the proteins functioning in solution were synthesized as soluble forms. This suggests the applicability of this protein synthesis method to determination of the solution structures of functional proteins. To examine this possibility, we have synthesized two (15)N labeled proteins and obtained (1)H-(15)N HSQC spectra for them. The structural analysis of these proteins has already progressed with an E. coli overexpression system, and (1)H-(15)N HSQC spectra for biologically active proteins have already been obtained. Comparing the spectra, we have shown that proteins synthesized with a wheat germ cell-free system have the proper protein folding and enough biological activity. This is the first experimental evidence of the applicability of the wheat germ cell-free protein synthesis system to high-throughput protein structural analysis. PMID- 12761394 TI - Automated protein fold determination using a minimal NMR constraint strategy. AB - Determination of precise and accurate protein structures by NMR generally requires weeks or even months to acquire and interpret all the necessary NMR data. However, even medium-accuracy fold information can often provide key clues about protein evolution and biochemical function(s). In this article we describe a largely automatic strategy for rapid determination of medium-accuracy protein backbone structures. Our strategy derives from ideas originally introduced by other groups for determining medium-accuracy NMR structures of large proteins using deuterated, (13)C-, (15)N-enriched protein samples with selective protonation of side-chain methyl groups ((13)CH(3)). Data collection includes acquiring NMR spectra for automatically determining assignments of backbone and side-chain (15)N, H(N) resonances, and side-chain (13)CH(3) methyl resonances. These assignments are determined automatically by the program AutoAssign using backbone triple resonance NMR data, together with Spin System Type Assignment Constraints (STACs) derived from side-chain triple-resonance experiments. The program AutoStructure then derives conformational constraints using these chemical shifts, amide (1)H/(2)H exchange, nuclear Overhauser effect spectroscopy (NOESY), and residual dipolar coupling data. The total time required for collecting such NMR data can potentially be as short as a few days. Here we demonstrate an integrated set of NMR software which can process these NMR spectra, carry out resonance assignments, interpret NOESY data, and generate medium-accuracy structures within a few days. The feasibility of this combined data collection and analysis strategy starting from raw NMR time domain data was illustrated by automatic analysis of a medium accuracy structure of the Z domain of Staphylococcal protein A. PMID- 12761395 TI - The long-range electrostatic interactions control tRNA-aminoacyl-tRNA synthetase complex formation. AB - In most cases aminoacyl-tRNA synthetases (aaRSs) are negatively charged, as are the tRNA substrates. It is apparent that there are driving forces that provide a long-range attraction between like charge aaRS and tRNA, and ensure formation of "close encounters." Based on numerical solutions to the nonlinear Poisson Boltzmann equation, we evaluated the electrostatic potential generated by different aaRSs. The 3D-isopotential surfaces calculated for different aaRSs at 0.01 kT/e contour level reveal the presence of large positive patches-one patch for each tRNA molecule. This is true for classes I and II monomers, dimers, and heterotetramers. The potential maps keep their characteristic features over a wide range of contour levels. The results suggest that nonspecific electrostatic interactions are the driving forces of primary stickiness of aaRSs-tRNA complexes. The long-range attraction in aaRS-tRNA systems is explained by capture of negatively charged tRNA into "blue space area" of the positive potential generated by aaRSs. Localization of tRNA in this area is a prerequisite for overcoming the barrier of Brownian motion. PMID- 12761393 TI - Why is Leu55-->Pro55 transthyretin variant the most amyloidogenic: insights from molecular dynamics simulations of transthyretin monomers. AB - Transthyretin (TTR) is one of the known human amyloidogenic proteins. Its native state is a homotetramer with each monomer having a beta-sandwich structure. Strong experimental evidence suggests that TTR dissociates into monomeric intermediates and that the monomers subsequently self-assemble to form amyloid deposits and insoluble fibrils. However, details on the early steps along the pathway of TTR amyloid formation are unclear, although various experimental approaches with resolutions at the molecular or residue level have provided some clues. It is highly likely that the stability and flexibility of monomeric TTR play crucial roles in the early steps of amyloid formation; thereby, it is essential to characterize initial conformational changes of TTR monomers. In this article we probe the possibility that the differences in the monomeric forms of wild-type (WT) TTR and its variants are responsible for differential amyloidogenesis. We begin with the simulations of WT, Val30-->Met (V30M), and Leu55-->Pro (L55P) TTR monomers. Nanosecond time scale molecular dynamics simulations at 300 K were performed using AMBER. The results indicate that the L55P-TTR monomer undergoes substantial structural changes relative to fluctuations observed in the WT and V30M TTR monomers. The observation supports earlier speculation that the L55P mutation may lead to disruption of the beta sheet structure through the disorder of the "edge strands" that might facilitate amyloidogenesis. PMID- 12761396 TI - Effects of sucrose on conformational equilibria and fluctuations within the native-state ensemble of proteins. AB - Osmolytes increase the thermodynamic conformational stability of proteins, shifting the equilibrium between native and denatured states to favor the native state. However, their effects on conformational equilibria within native-state ensembles of proteins remain controversial. We investigated the effects of sucrose, a model osmolyte, on conformational equilibria and fluctuations within the native-state ensembles of bovine pancreatic ribonuclease A and S and horse heart cytochrome c. In the presence of sucrose, the far- and near-UV circular dichroism spectra of all three native proteins were slightly altered and indicated that the sugar shifted the native-state ensemble toward species with more ordered, compact conformations, without detectable changes in secondary structural contents. Thermodynamic stability of the proteins, as measured by guanidine HCl-induced unfolding, increased in proportion to sucrose concentration. Native-state hydrogen exchange (HX) studies monitored by infrared spectroscopy showed that addition of 1 M sucrose reduced average HX rate constants at all degrees of exchange of the proteins, for which comparison could be made in the presence and absence of sucrose. Sucrose also increased the exchange-resistant core regions of the proteins. A coupling factor analysis relating the free energy of HX to the free energy of unfolding showed that sucrose had greater effects on large-scale than on small-scale fluctuations. These results indicate that the presence of sucrose shifts the conformational equilibria toward the most compact protein species within native-state ensembles, which can be explained by preferential exclusion of sucrose from the protein surface. PMID- 12761397 TI - Structural perturbation and enhancement of the chaperone-like activity of alpha crystallin by arginine hydrochloride. AB - Structural perturbation of alpha-crystallin is shown to enhance its molecular chaperone-like activity in preventing aggregation of target proteins. We demonstrate that arginine, a biologically compatible molecule that is known to bind to the peptide backbone and negatively charged side-chains, increases the chaperone-like activity of calf eye lens alpha-crystallin as well as recombinant human alphaA- and alphaB-crystallins. Arginine-induced increase in the chaperone activity is more pronounced for alphaB-crystallin than for alphaA-crystallin. Other guanidinium compounds such as aminoguanidine hydrochloride and guanidine hydrochloride also show a similar effect, but to different extents. A point mutation, R120G, in alphaB-crystallin that is associated with desmin-related myopathy, results in a significant loss of chaperone-like activity. Arginine restores the activity of mutant protein to a considerable extent. We have investigated the effect of arginine on the structural changes of alpha-crystallin by circular dichroism, fluorescence, and glycerol gradient sedimentation. Far-UV CD spectra show no significant changes in secondary structure, whereas near-UV CD spectra show subtle changes in the presence of arginine. Glycerol gradient sedimentation shows a significant decrease in the size of alpha-crystallin oligomer in the presence of arginine. Increased exposure of hydrophobic surfaces of alpha-crystallin, as monitored by pyrene-solubilization and ANS-fluorescence, is observed in the presence of arginine. These results show that arginine brings about subtle changes in the tertiary structure and significant changes in the quaternary structure of alpha-crystallin and enhances its chaperone-like activity significantly. This study should prove useful in designing strategies to improve chaperone function for therapeutic applications. PMID- 12761398 TI - Protein-protein docking with a reduced protein model accounting for side-chain flexibility. AB - A protein-protein docking approach has been developed based on a reduced protein representation with up to three pseudo atoms per amino acid residue. Docking is performed by energy minimization in rotational and translational degrees of freedom. The reduced protein representation allows an efficient search for docking minima on the protein surfaces within. During docking, an effective energy function between pseudo atoms has been used based on amino acid size and physico-chemical character. Energy minimization of protein test complexes in the reduced representation results in geometries close to experiment with backbone root mean square deviations (RMSDs) of approximately 1 to 3 A for the mobile protein partner from the experimental geometry. For most test cases, the energy minimized experimental structure scores among the top five energy minima in systematic docking studies when using both partners in their bound conformations. To account for side-chain conformational changes in case of using unbound protein conformations, a multicopy approach has been used to select the most favorable side-chain conformation during the docking process. The multicopy approach significantly improves the docking performance, using unbound (apo) binding partners without a significant increase in computer time. For most docking test systems using unbound partners, and without accounting for any information about the known binding geometry, a solution within approximately 2 to 3.5 A RMSD of the full mobile partner from the experimental geometry was found among the 40 top scoring complexes. The approach could be extended to include protein loop flexibility, and might also be useful for docking of modeled protein structures. PMID- 12761399 TI - Importance of alpha-helix N-capping motif in stabilization of betabetaalpha fold. AB - FSD-1 (full sequence design 1) is a protein folded in a betabetaalpha motif, designed on the basis of the second zinc finger domain of Zif268 by a substitution of its metal coordination site with a hydrophobic core. In this work, we analyzed the possibility of introducing the DNA recognition motif of the template zinc finger (S(13)RSDH(17)) into FSD-1 sequence in order to obtain a small DNA-binding module devoid of cross-link(s) or metal cofactors. The hybrid protein was unfolded, as judged by CD and NMR criteria. To reveal the role of each of the five amino acids, which form the N-capping motif of the alpha-helix, we analyzed conformational and stability properties of eight FSD-1 mutants. We used a shielded methyl group of Leu 18 and a CD signal at 215 nm as a convenient measure of the folded state. Glu 17-->His substitution at the N(3) in S(13)NEKE(17) variant decreased the folded structure content from 90% to 25% (equivalent to 1.8 kcal * mole(-1) destabilization) by disruption of N-capping interactions, and had the most significant effect among single mutants studied here. The N(cap) Asn 14 substitution with Arg considerably decreased stability, reducing structure content from 90% to 40% (1.4 kcal * mole(-1) destabilization) by disruption of a helix-capping hydrogen bond and destabilization of a helix macrodipole. The N(1) Glu 15-->Ser mutation also produced a considerable effect (1.0 kcal * mole(-1) destabilization), again emphasizing the significance of electrostatic interactions in alpha-helix stabilization. PMID- 12761400 TI - Analysis of transglutaminase protein substrates by functional proteomics. AB - Transglutaminases are calcium-dependent enzymes that catalyze a post translational modification of proteins through the formation of epsilon -(gamma glutamyl)lysine bonds. Although specific roles for transglutaminases have been described, recent findings have provided evidence that dysregulation of transglutaminases may contribute to many pathological processes including celiac disease and neurodegenerative diseases. A crucial step in the elucidation of biological and pathological roles of transglutaminases requires the identification of protein substrates. A strategy based on a functional proteomic analysis was set up using two well-characterized biotinylated transglutaminase substrates as affinity probes: 5-(biotinamido)pentylamine and the synthetic biotinylated peptide TVQQEL, the amino- and acyl-donor probes, respectively. A pool of known tissue type transglutaminase protein substrates was selected in order to test the procedure. Results obtained in this paper indicate that the whole strategy can be successfully applied in order to identify transglutaminases protein substrates as well as the amino acid site sensitive toward enzyme activity. PMID- 12761401 TI - Propensities, probabilities, and the Boltzmann hypothesis. AB - The relative strengths of interactions involving polypeptide chains can be estimated with reasonable accuracy with statistical potentials, free-energy functions derived from the frequency of occurrence of structural arrangements of residues or atoms in collections of protein structures. Recent published work has shown that the energetics of side-chain/backbone interactions can be modeled by the phi/psi propensities of the 20 amino acids. In this report, the more commonly used phi/psi probabilities are demonstrated to fail in evaluating the free energies of protein conformations because of an overriding preference for all helical structures. Comparison of the hypothetical reactions implied by these two different statistics-propensities versus probabilities-leads to the conclusion that the Boltzmann hypothesis may only be applicable for the calculation of statistical potentials after the starting conformation has been specified. This conclusion supports a simple conjecture: The surprising success of the Boltzmann hypothesis in explaining the energetics of protein structures is a direct consequence of a real equilibrium, one extending over evolutionary time that has maintained the stability of each protein within a narrow range of values. PMID- 12761402 TI - The Zintl-Klemm concept applied to cations in oxides. I. The structures of ternary aluminates. AB - The structures of 94 ternary aluminates are reinterpreted on the basis of the Zintl-Klemm concept and Pearson's generalized octet rule. In aluminates of highly electropositive metals such as alkali, alkaline-earth and rare-earth metals, the Al atoms form three-dimensional skeleta which can be interpreted as if the Al atoms were behaving as Zintl polyanions, adopting the structure of either main group elements or Zintl polyanions showing the same connectivity. The O atoms are then located close to both the hypothetical two-electron bonds and the lone pairs, giving rise to a tetrahedral coordination. When more electronegative elements, such as W or Si, are present in the compound, the electron transfer towards the Al atoms does not take place. In this case, aluminium behaves as a base, transferring its electrons to the more electronegative atoms and the coordination sphere of aluminium becomes octahedral. In some compounds the Al atoms clearly show amphoteric character so that some Al atoms act as donors (bases) and hence are octahedrally coordinated, whereas others behave as acceptors (acids), adopting a tetrahedral coordination. From this it is concluded that the coordination sphere of aluminium is not a function of the ionic radius of the Al(3+) cations, but it depends on the nature of the other cations accompanying them in the structure. The networks formed by these aluminates are, in many instances, similar to those of the binary oxides of the main-group elements. For this reason, a systematic survey of these oxides is also reported. Compounds such as stuffed cristobalites and trydimites and also perovskites are examples of this new interpretation. Perovskites are then reinterpreted as a stuffed pseudo-TeO(3) structure. Other families of compounds such as silicates and phosphates are susceptible to a similar interpretation. This study provides additional examples of how cations recognize themselves in spite of being embedded in an oxygen matrix. PMID- 12761403 TI - The commensurate composite sigma-structure of beta-tantalum. AB - The single-crystal investigation of the self-hosting sigma-structure of beta tantalum (beta-Ta) at 120 K (low-temperature, LT, structure) and at 293 K (RT-I before cooling and RT-II after cooling and rewarming; RT represents room temperature) shows that this structure is indeed a specific two-component composite where the components have the same (or an integer multiple) lattice constants but different space groups. The space groups of both host (H) and guest (G) components cause systematic absences, which result from their intersection. The highest symmetry of a sigma-structure can be described as [H: P4(2)/mnm; G: P4/mbm (c(G) = 0.5c(H)); composite: P4(2)/mnm]. A complete analysis of possible symmetries is presented in the Appendix. In beta-Ta, two components modify their symmetry during the thermal process 293 K (RT-I)=> 120 K (LT)=> 293 K (RT-II): [H: P(-)42(1)m; G: P(-)42(1)m; composite: P(-)42(1)m]=> [H: P(-)4, G: P4/mbm (c(G) = 0.5c(H)), composite: P(-)4]=> [H: P(-)42(1)m, G: P4/mbm (c(G) = 0.5c(H)), composite: P(-)42(1)m]. Thus, the phase transition is reversible with respect to H and irreversible with respect to G. PMID- 12761404 TI - Sodium carbonate revisited. AB - We present the structure of anhydrous sodium carbonate at room temperature (phase gamma) and 110 K (phase delta) based on single-crystal X-ray diffraction data. The incommensurate phase gamma was determined almost 30 years ago in the harmonic approximation using one modulation wave and first-order satellites. In our work we use satellites up to fifth order and additional harmonic waves to model the anharmonic features of the structure. The commensurate phase delta is presented for the first time. Using the superspace approach, both phases are compared in order to find common trends in the whole range of the sodium carbonate phases. We present arguments supporting the hypothesis that the driving force of the phase transitions may originate in the unsaturated bonding potential of one of the Na ions. PMID- 12761405 TI - Cation movement and phase transitions in KTP isostructures; X-ray study of sodium doped KTP at 10.5 K. AB - An accurate structure model of sodium-doped potassium titanyl phosphate, (Na(0.114)K(0.886))K(TiO)(2)(PO(4))(2), has been determined at 10.5 K by single crystal X-ray diffraction. In addition to the low-temperature data, X-ray intensities have been collected at room temperature. When the temperature was decreased from room temperature to 10.5 K, both potassium cations moved 0.033 (2) A along the c-axis, i.e. in the polar direction within the rigid Ti-O-P network. This alkaline metal ion displacement can be related to the Abrahams-Jamieson Kurtz T(C) criteria for oxygen framework ferroelectrics. Potassium titanyl phosphate (KTP) is a well known material for second harmonic generation (SHG), and the influence of sodium dopant on the TiO(6) octahedral geometry and SHG is discussed. The material studied crystallizes in the space group Pna2(1) with Z = 4, a = 12.7919 (5), b = 6.3798 (4), c = 10.5880 (7) A, V = 864.08 (9) A(3), T = 10.5 (3) K and R = 0.023. PMID- 12761407 TI - Structure of lithium benzilate hemihydrate solved by simulated annealing and difference Fourier synthesis from powder data. AB - The crystal structure of lithium benzilate hemihydrate (C(14)H(11)O(3)( )Li(+).0.5H(2)O) was solved from synchrotron powder diffraction data. This compound crystallizes in the monoclinic space group P2(1)/a. The structure was solved via the direct space search for two benzilate fragments using the simulated-annealing program DASH, localization of the lithium ions and water molecule from a difference Fourier map, and a restrained Rietveld refinement (R(wp) = 0.0687). The structure is a coordination polymer of [Li(2)(C(14)H(11)O(3))(2).H(2)O](2) tetramers building helical fourfold one dimensional channels parallel to [010]. Inside the channels the tetrahedral coordination spheres of the lithium ions contain hydroxyl and carbonyl groups, and water molecules. The water molecule functions as the cohesive entity forming extended hydrogen-bonded chains running along [010], and bifurcated donor hydrogen bonds with the two nearest carboxylates. At the outer edge of the channels, weaker intermolecular C-H.Ph hydrogen bonds along [100] and [001] contribute to the supramolecular aggregation of the structure. PMID- 12761409 TI - Redetermination of the crystal structure of alpha-copper phthalocyanine grown on KCl. AB - The crystal structure of a polymorph of copper phthalocyanine (CuPc) grown on a KCl substrate is redetermined by transmission electron diffraction. It has a triclinic unit cell containing one molecule; the crystal does not have a herringbone-type molecular arrangement, which is a common packing mode of planar phthalocyanines. The molecular packing is determined by the diffraction intensity with the aid of the calculation of molecular packing energy. One of the striking features of this polymorph is its stacking mode within a molecular column: the molecular stacking direction projected on a molecular plane is different by an angle of about 45 degrees from that of the alpha-modifications of platinum phthalocyanine (PtPc) and metal-free phthalocyanine (H(2)Pc). A powder X-ray diffraction profile calculated for the polymorph agrees well with that of so called alpha-CuPc and Rietveld analysis for alpha-CuPc indicates that the CuPc crystals grown on KCl are actually alpha-CuPc; hence, alpha-CuPc is not isostructural with either alpha-PtPc or alpha-H(2)Pc. On the basis of the present results and the reported crystal structures of the planar phthalocyanines that form molecular columns, the polymorphs of the phthalocyanines can be classified into four types distinguished by the molecular stacking mode within the column: alpha(x)-, alpha(+)-, beta(x)- and beta(+)-types. PMID- 12761406 TI - Structure-forming components in crystals of ternary and quaternary 3d-metal complex fluorides. AB - Crystallochemical analysis and classification were performed for 139 ternary and quaternary complex fluorides with the general formula M1(n)M2(m)M3F(6), belonging to 33 structure types. Using coordination sequences and the uniformity criterion the structure-forming ionic sublattices or their combinations were found, which are responsible for the formation of stable periodic frameworks. Analysis of structure-forming motifs allows the interpretation of the crystal structures of complex fluorides as close packings of F ions with M1, M2 and M3 cations, partially occupying tetrahedral and octahedral voids, or as the packings of [M3F(6)] complex ions with M1 and M2 countercations in the voids. Cationic sublattices are noted to play an essential role, while forming crystal structures of complex fluorides. Relationships between the composition of structure-forming sublattices, the composition of compounds, and the size and charge of ions belonging to the sublattices were analysed under normal conditions, with thermal and high-pressure polymorphic transitions. Rules were formulated to predict the crystal structures of complex fluorides with a given chemical composition. PMID- 12761408 TI - Diffuse scattering and short-range order in uranium iodine phthalocyanine [U1 xPc(2)]I2-y and the X-ray structure analysis of crystals with diffuse superstructure reflections. AB - Crystals of uranium iodine phthalocyanine present an example of a disordered commensurate modulated structure of the intergrowth type. The short-range order of both uranium ions and iodine chains [I(3)(-)](n) has been analysed by Reverse Monte Carlo (RMC) simulation of X-ray diffuse scattering. The diffraction pattern of uranium iodine phthalocyanine contains diffuse superstructure reflections. In the routine crystal structure analysis diffuse superstructure reflections may be either omitted or measured and classified along with other Bragg reflections. The crystal structure of uranium iodine phthalocyanine is an example of such ambiguity. The crystal structures of two specimens of [U(1-x)Pc(2)]I(2-y) with slightly different composition have been published in the literature with different space groups and unit cells. We have shown that the structure of both specimens differs only in the degree of short-range order and is isostructural with [YbPc(2)]I(2). We have also shown that while the omission of diffuse reflections results in the average crystal structure, the treatment of these reflections as normal Bragg reflections is incorrect and produces the structure averaged over a limited small range. PMID- 12761410 TI - Mechanism of the first-order phase transition of an acylurea derivative: observation of intermediate stages of transformation with a detailed temperature resolved single-crystal diffraction method. AB - The process of the first-order solid-to-solid phase transition of 1-ethyl-3-(4 methylpentanoyl)urea (1) was observed by means of a detailed temperature-resolved single-crystal diffraction method, which resembles watching a series of stop motion photographs. The transition consists of two elementary processes, one supramolecular and the other molecular. Crystal structures from before and after the phase transition are isostructural. The straight-ribbon-like one-dimensional hydrogen-bonding structure is formed and stacked to form a molecular layer. The geometry of the layer is retained during the phase transition. The relative position of the layer with its neighbours, on the other hand, changes gradually with increasing temperature. The change is accelerated at the temperature representing the start of the endotherm seen in the DSC curves of (1). The structural variation yields void space between the neighbouring layers. When the void space grows enough that the crystal is unstable, the 3-methylbutyl group on the last of the molecules turns into a disordered structure with drastic conformational changes to fill up the void space. The phase transition process is well supported with simple force-field calculations. A crystal of 1-(4 methylpentanoyl)-3-propylurea (2), which shows no solid-to-solid phase transitions, was also analysed by the same method for comparison. PMID- 12761411 TI - Structural changes of hexamethylenetetramine and undecanedioic acid co-crystal (HMT-C11) as a function of the temperature. AB - HMT-C11 belongs to the family of adducts formed by the co-crystallization of N(4)(CH(2))(6) molecules (hexamethylenetetramine or HMT) and aliphatic dicarboxylic acids HOOC(CH(2))(n - 2)COOH with 5 or =3 g/l) (p=0.005), expressed glomerulosclerosis (p=0.005) and expressed interstitial fibrosis (p=0.034). Focal segmental glomerulosclerosis were found to have a relatively bad long-term prognosis--the renal survival rate in 5 years was 77.8%. Kaplan-Meier survival analysis showed that expressed glomerulosclerosis was risk factor (logrank p=0.016, Breslov p=0.043) associated with end-stage renal disease in 5 years. PMID- 12761416 TI - [Diagnosis of renal vascular disease]. AB - The aim of the study was to assess the diagnostic value of gamma scintigraphy, duplex ultrasound scan, computer tomographic angiography (CTA) and renal arteriography in 77 patients with arterial hypertension, 3 with hematuria, 3 with chronic renal insufficiency and 3 with the difference in kidney size. The following kidney vascularization pathology was revealed: renal artery stenosis in 27, thrombosis of renal artery in 1, aneurysm of renal artery in 4, arteriovenous fistulae in 1, anomalies of renal arteries in 12, atherosclerotic deterioration of accessorial renal arteries in 13, tumor of suprarenal gland with arterial stenosis in 2 patients. Gamma scintigraphy and duplex ultrasound scan were positive in detecting vascular pathology in 93.7% and 66.7% patients respectively. The most informative methods were CTA and renal angiography. CTA failed to assess the grade of renal artery stenosis in 5 patients. PMID- 12761418 TI - [Rapidly progressive glomerulonephritis syndrome: course, pathomorphology and outcome (data of Kaunas University of Medicine Hospital 1996-2002)]. AB - Rapidly progressive glomerulonephritis is rare but severe clinical syndrome, which results in end-stage renal disease if not treated. The prognosis can be improved with early diagnosis and aggressive treatment. The aim of our study was to analyze retrospectively all cases of rapidly progressive glomerulonephritis, diagnosed in Kaunas University of Medicine Hospital since 1996 and to evaluate the clinical, laboratory, immunological, pathomorphological data, treatment and outcome in these patients. In period of 1996-2002, 23 patients were treated in Departments of Nephrology, Rheumatology, and Pediatric Nephrology. Rapidly progressive glomerulonephritis was observed in elderly persons, the mean age 46.48+/-5.28 years. In 21.74% of cases, patients were diagnosed Goodpasture syndrome, in 21.74% of cases immune complex disease and in 56.52%--pauci immune glomerulonephritis which was mainly limited in kidney as necrotizing crescentic glomerulonephritis. Rapidly progressive glomerulonephritis in most cases manifested by weakness, hypertension, fever, involvement of lower airways, infection. Renal biopsies were performed in 18 (78.26%) cases, lung biopsy was made for one patient. Biopsies were made on 30.56+/-7.41 day of hospitalization. Twenty-two (95.65%) patients received intravenous pulses with methylprednisolone and/or cyclophosphamide; 56.52% patients required dialysis. Renal function improved in 47.83% patients; 8 (34.78%) patients died. The causes of death were infections complications, progression of main disease and cardiac arrest. PMID- 12761417 TI - [Potential causes of antigenemia in the patients with an immune complex-mediated membranoproliferative glomerulonephritis in Lithuania]. AB - The pathogenesis of an immune complex-mediated membranoproliferative glomerulonephritis (IMPGN) involves persistent deposition of circulating immune complexes in the glomeruli caused by persistent antigenemia. We have previously reported relatively high incidence of IMPGN in Lithuania. The objective of our study was to evaluate potential causes of persistent antigenemia in the patients with IMPGN. MATERIAL AND METHODS: Forty-five patients with IMPGN diagnosed on renal biopsy during 2000-2002 were retrospectively evaluated for the presence of persistent bacterial or viral infections, autoimmune diseases and other associated medical conditions. Patients with established diagnosis of systemic lupus erythematosus (SLE) before the biopsy were not included in the study. RESULTS: A great majority (20; 44%) of the patients were found to have persistent bacterial infections of various localization. Four patients (9%) were infected with hepatitis B virus (HBV). Three (7%) patients were eventually diagnosed with SLE and another 3 (7%) had other associated pathology. In the remaining 15 (33%) patients, IMPGN remained idiopathic. Testing for hepatitis C virus (HCV) antibody was performed in 36 patients (12 of them with idiopathic IMPGN) and was negative in all patients. Testing for HCV RNA was not performed. Patients with bacterial infections were significantly younger compared to the group of idiopathic IMPGN (36.5+/-19.1 and 53.8+/-16.4, respectively, p=0.01). We conclude that persistent bacterial infection was a major potential source of antigenemia in our patients with IMPGN, particularly in the younger patients, while HBV and HCV infection was rarely detected. PMID- 12761419 TI - [Clinical and laboratory features and prognostic implications in myeloma with and without renal impairment]. AB - Presenting clinical and laboratory features, prognostic implications and survival in 124 multiple myeloma patients were reviewed in a retrospective study based on hospital records. The median age was 65 years. Out of all patients, 2.42% were younger than 40 years and 62.9% were 60 years and older. The main presenting clinical features were bone pain (70.16%), fatigue (31.45%), recurrent infections (9.68%) and weight loss (0.3%). Renal failure was present in 35.48% of patients. The higher means of ionised calcium, uric acid, erythrocyte sedimentation rate, M protein were correlated with the higher mean of serum creatinine. The acturial survival of myeloma patients without renal failure at 1 and 2 years was 95.08% and 89.23% respectively, while acturial survival of myeloma patients with renal failure at 1 and 2 years was 82.5% and 73.35% respectively (p<0.01). One-year survival in myeloma patients maintained on chronic hemodialysis was 68.75% while it is reported as 90.91% for myeloma patients not on dialysis (p<0.006). PMID- 12761420 TI - [Renal failure due to a villous adenoma]. AB - Villous adenoma comprises 5.6% of colon tumors. Adenomas of the colon could be divided into tubular, villous, and tubulovillous adenomas. Villous adenomas are not as common as tubular neoplasms; however, usually they are larger and could cause a depleting syndrome characterized by dehydration, hyponatremia, hypokalemia, hypochloremia, and metabolic acidosis. In severe cases, shock and renal failure in older patients could cause death. Case of villous adenoma is presented in this article. Seventy-eight-year-old male who suffered permanent diarrhea for few years followed with acute renal failure. Villous adenoma was diagnosed. The conservative treatment had short renal function improvement. Hemodialysis had to be performed. Renal function was completely recovered after radical tumor surgery. PMID- 12761421 TI - [Dynamics of diabetic nephropathy and other complications of type I diabetes mellitus in the period of 1996-2002 (data from 2 Kaunas outpatient polyclinics)]. AB - Kidney involvement in diabetes mellitus has negative impact on the outcomes of disease. Strong relationship between progressive diabetic kidney disease and the development of other diabetic complications was found by many investigators. In order to evaluate the dynamics of diabetic nephropathy in type I diabetes mellitus during 6-year period and its relationship with other diabetes mellitus complications and control of glycemia and hypertension, in 2002 we reviewed ambulatory case records of patients, who were followed by endocrinologists and who were investigated by us in 1996. During 6-year period, 5.1% from 156 pts. died and all of them had diabetic nephropathy; 26.9% of pts. moved to general practitioners and never visited endocrinologists again. Only 105 pts. remained under follow-up by endocrinologists. Their mean age 37.6+/-1.3 yrs. Out of all patients, 54% were males and 46% females. Mean diabetes mellitus duration was 19.5+/-0.9 yrs. Control of glycaemia was poor and insufficient in 2/3 of pts. HbA(1C) wasn't checked in 68.9% of pts. Control of arterial hypertension became better, but not sufficiently. During 6-year period persistent proteinuria developed in 12.1% of pts., who had no or transient proteinuria <0.5 g/l in 1996. Persistent proteinuria developed 19.9+/-1.8 yrs. after the diabetes mellitus onset and correlated with hypertension and renal insufficiency. Higher level of proteinuria was associated with worse control of glycemia. Progression of diabetic retinopathy and neuropathy over 6 yrs. were more expressed than in diabetic nephropathy. On average retinopathy developed after 14+/-1.8 yrs. after the diabetes mellitus onset, neuropathy--17.8+/-2.2 yrs., renal failure--21.1+/ 2.8 yrs., heart failure--22.9+/-1.9 yrs. and arterial hypertension--12.1+/-1.3 yrs. The prevalence and time of incipient diabetic nephropathy appearance remained unknown because the test for microalbuminuria was not available in the primary health care centres. PMID- 12761422 TI - [Long-term Cefadroxil prophylaxis in children with recurrent urinary tract infections]. AB - The aim of the study was to evaluate the efficacy and safety of low-dose, long term cefadroxil prophylaxis in preventing recurrent urinary tract infections in children. A prospective, randomized trial in 33 children (32 female, 1 male) aged 2-14 years (mean 8.1+/-2.8) was conducted. Children with recurrent urinary tract infections were commenced to six-month prophylaxis with cefadroxil at the dosage of 12.5-15.0 mg/kg at bedtime administered every night (group I, n=15) or alternate night (group II, n=18). Escherichia coli was the most frequently isolated agent (90.9%), followed by Proteus mirabilis (3.0%) and Klebsiella oxytoca (3.0%) and Staphylococcus epidermidis (3.0%). Cefadroxil prophylaxis was effective in 80% of patients in group I and in 78% patients in group II. Reinfection occurred in 3/15 (20%) patients in-group I and in 4/18 (22.2%) patients in-group II (odds ratio--1.14; 95% confidence interval--0.16-8.3). The difference of reinfection rate between the groups was not significant (p=0.88). The rate of day- and night-time wetting in both groups before prophylaxis of urinary tract infections was high. It decreased significantly at the end of prophylactic treatment with cefadroxil (from 42.4% to 9.1% and from 39.4% to 6.1% respectively). Mild adverse reaction (nausea) was observed in 1/33 patient. In conclusion our study shows that cefadroxil is an effective, well-tolerated and safe agent in the urinary tract infections prophylaxis. Prophylaxis of recurrent urinary tract infections in children with cefadroxil on alternate night regimen might reduce the cost of the treatment. PMID- 12761423 TI - [Differential diagnosis of nephritic and nephrotic syndroms in children]. AB - The aim of this study was to compare clinical and laboratory features of children with acute poststreptococcal glomerulonephritis and steroid sensitive nephrotic syndrome. We have examined 30 children with acute poststreptococcal glomerulonephritis and 17 children with steroid sensitive nephrotic syndrome, who arrived for hospitalization at Kaunas University of Medicine Hospital in 1997 2000. All patients not later than in ten days from the onset of acute glomerulonephritis and 14 days later were given a standartized clinical and laboratorical examination following a record. We determined that hematuria is universal finding of acute poststreptococcal glomerulonephritis and proteinuria was found in all patients with steroid sensitive nephrotic syndrome. Microscopic hematuria and proteinuria were persisted in children who had acute poststreptococcal glomerulonephritis. Anemia was found only in patients who were ill with acute poststreptococcal glomerulonephritis. After 14 days of treatment it was determined in 38.5% of children. Hypoproteinemia was established in 69.2% of children and hypercholesterolemia in 92.3% of children with steroid sensitive nephrotic syndrome after two weeks of treatment. Decreased glomerular filtration rate was characteristic feature of acute poststreptococcal glomerulonephritis. PMID- 12761424 TI - [Incidence of peritonitis and catheter exit site infection in children undergoing automatic continuous cyclic peritoneal dialysis]. AB - The aim of this study was to find out the clinical aspects of peritonitis and catheter exit site infection in children undergoing continuous cyclic peritoneal dialysis. The incidence of peritonitis and catheter related infections were reviewed in 8 children on continuous cyclic peritoneal dialysis over a mean period of 14.9+/-15.8 months. Peritonitis occurred in 4 children. There were 14 episodes of peritonitis. The mean time from starting dialysis to the first episode of peritonitis was 1.9+/-1.0 months. The incidence of peritonitis was 1 episode in 9 treatment months. Gram-negative organisms (Enterococcus spp.) were responsible for the majority of episodes (35.7%) of peritonitis and gram-positive bacteria mainly caused the catheter exit site infections (Staphylococcus epidermidis). The incidence of catheter exit site infection was one episode in 8 treatment months. Recurrent peritonitis was present in 1 case. Most patients with dialysis-related peritonitis and catheter exit site infection responded to antibiotic therapy. Catheter was replaced in 2 patients. The mortality rate was 2 out of 8 patients but none of the deaths were related to peritonitis. Patients with exit site infection had 7 times higher risk than those without developing peritonitis. PMID- 12761425 TI - [Changes of etiology and management of childhood urinary tract infection during the period of 1994-2002 (data of Kaunas University of Medicine Hospital)]. AB - The purpose of this article was to establish changes of age, etiology, diagnostics and treatment for children, who were treated for urinary tract infection in Clinic of Children Diseases, Kaunas University of Medicine Hospital in 1994-1998 and 1999-2002. Seven hundred fifty children urinary tract infections were treated during the period of 1994-1998, and 673--during the period of 1999 2002. Female patients had urinary tract infection nearly five times more than male patients in other age groups. Rate of urinary tract infection in male patients under one year age was greater than in female in this age group (p=0.0001). Greater incidence of urinary tract infection in age group under one year age was established. The incidence among male patients with urinary tract infection grew from 37.8% to 64.0% (p=0.0001) and among female patients--from 9.6% to 18.1% (p<0.0001). More vesicoureteral refluxes (VUR) were diagnosed: 44 (7.8%) in period 1994-1998 and 52 (9.0%) in the period 1999-2002. More than one third of all VUR (36.5%) were diagnosed in age group under one year. The incidence of VUR in this group was greater for male than for female patients (p=0.015). More positive urinary culture for patients with pyelonephritis were in period 1999-2002 (74.6%) than in 1994-1998 (66.8%). The incidence of microorganisms was nearly the same. E. coli was found in 83.4% and in 83.9% of all positive urinary cultures. Sensitivity to the commonly used antibiotics was better (p<0.0001). Pyelonephritis mostly was treated with gentamycin. PMID- 12761426 TI - [Influence of omega-3 fatty acids on lipid metabolism in children with steroid sensitive nephrotic syndrome]. AB - Changes in lipid metabolism is one of the most important features of steroid sensitive nephrotic syndrome. Increased serum lipid level not always is connected with disease activity and sometimes may persist for long time, especially in frequently relapsing nephrotic syndrome. Hyperlipidemia and dyslipidemia are connected with cardiovascular diseases and development of glomerulosclerosis. Omega-3 (omega-3) polyunsaturated fatty acids have positive influence on lipid metabolism, diminishing hypercoagulation and a risk of cardiovascular diseases. The aim of this study was to examine influence of omega-3 polyunsaturated fatty acids on lipid disturbances in children with steroid sensitive nephrotic syndrome. Forty-eight patients were included in the study; 24 of them were treated with prednisolone (control group) and 24 were treated with prednisolone and fish oil (6 mg/day) along with symptomatic therapies. Serum lipid profile and spectrum of fatty acids before treatment and three months after start of the treatment were evaluated. Before treatment, contents of serum fatty acids and lipids did not differ between the groups. The study showed decrease in serum triglycerides (p<0.06) and omega-6 polyunsaturated fatty acids (p<0.02), increase in concentration of omega-3 polyunsaturated fatty acids and the ratio of 20:5 omega-3/20: omega-6 (eicosapentaenoic/arachidonic acids) (p<0.001). Cholesterol, HDL-Ch, LDL-Ch concentrations did not change under the treatment with fish oil. We conclude that fish oil might be useful as an agent normalising lipid metabolism in children with steroid sensitive nephrotic syndrome. PMID- 12761427 TI - [Bartter syndrome and it's neonatal type]. AB - For the first time neonatal variant of Bartter syndrome to 14.5-year old girl is presented in Lithuania. It is a rare genetical disease with autosomal recessive inheritance. The patient was born prematurely, had polyhydramnion, polyuria and polydypsia, a craving for salt, specific outlook and was mentally retarded, had muscle weakness and nephrocalcinosis. Hypokalemia, hyperreninemia and metabolic alkalosis were found. Urine analysis revealed impaired renal concentration capacity, hypercalciuria and hypernatriuria. She had the symptom of systemic disease - osteopenia. Literature review on Bartter's syndrome is done. PMID- 12761429 TI - [Water treatment systems of hemodialysis centers in Lithuania and trace metals in purified water in 2002]. AB - The objective of this survey was to obtain information on hemodialysis chemical water quality and on water treatment systems of hemodialysis centers in Lithuania. Five trace metals (Al, Pb, Cd, Zn, Cu) were examined in the purified water (sample from a point after the water treatment system) of 28 hemodialysis centers. Atomic absorption spectrophotometry was applied to measure water trace metals levels. All hemodialysis centers in Lithuania used treated water. Softeners were used by 100%, reverse osmosis by 86.2% of the centers. Concomitant use of sand filter, softeners, activated carbon, reverse osmosis was found in 72.4% of the centers. The age of the water treatment system varied from 1 to 117 months (mean=39.7+/-30.4). Concentrations of Al, Pb, Cd, Zn, Cu in the purified water of 28 hemodialysis centers did not exceed standards of the European Pharmacopoeia. There was significant decrease in the mean levels of investigated trace elements in the treated water in Lithuania in 2002 compared with examined in 1998. PMID- 12761428 TI - [Urinary microflora in children from different pediatric departments]. AB - For rational treatment of urinary tract infection it is necessary to know the causative agents. These may be different in various departments of health care institution managing patients with different illnesses. The article presents results of urinary cultures performed in 2002 in Microbiology Laboratory of Vilnius University Children's Hospital. The urine specimens were sent to the Laboratory from all departments of the Hospital. The most common isolate in all departments was E. coli (63.1% of all cultures with growth of >/=10(5) cfu/ml). This organism was somewhat less common in urine specimens obtained from Department of Urology (54.8%); accordingly, species of other organisms were isolated somewhat more often. S. aureus most often was identified in urine specimens from neonatal departments (11.1% of all significant isolates). There were no significant differences in urinary microflora isolated from pediatric in patients and out-patients. Majority of the strains of the main urinary pathogen, E. coli, showed susceptibility to gentamicin, II-III generation cephalosporins, co-amoxiclav, nitrofurantoin and ciprofloxacin. Thus for empirical treatment of urinary tract infection it is necessary to choose an antibiotic with good activity against E. coli. In more complex circumstances (e.g. in children with urinary tract anomalies and in those treated in intensive care units) it would be better to take into account the results of urinary culture and susceptibility testing. PMID- 12761430 TI - [Aluminum concentration in blood of hemodialysis patients and its clinical importance]. AB - A multicenter study was performed to investigate the prevalence of abnormal blood contents of aluminum in hemodialysis patients and to clarify the impact of aluminum level on the clinical status of such patients. MATERIAL AND METHODS: Two hundred sixty five patients with end-stage renal disease from 7 dialysis centers were enrolled in this study. All patients had undergone standard hemodialysis. Venous blood samples were collected from hemodialysis patients before hemodialysis sessions. Atomic absorption spectrophotometry was applied to measure blood levels of aluminum. RESULTS: Out of hemodialysis patients 24.9% had high blood aluminum (>30 microg/l), and the mean was 27.4+/-43.8 microg/l in all subjects. The aluminum containing phosphate binder users had significantly higher blood aluminum levels (45.1+/-102.0 versus 15.4+/-18.9 microg/l, p<0.04). The hemodialysis subjects had low parathyroid hormone level more frequently when blood aluminum concentration was high (>30 microg/l). PMID- 12761431 TI - [Hospitalizations of hemodialysis patients (data of Kaunas University of Medicine Hospital in 2001)]. AB - The number of patients with end-stage renal disease is increasing, therefore the need of renal replacement therapy is increasing. Rising cost of treating end stage renal disease patients includes not only the cost of dialysis procedure but also the cost of hospitalizations and ambulatory medicamental treatment. There is no analysis done of hospitalizations of end-stage renal disease patients in Lithuania. Our purpose--to examine hospitalization rate, causes and duration in end-stage renal disease patients dialyzed in Kaunas University of Medicine Hospital. We analyzed ambulatory case records of all end-stage renal disease patients dialyzed in our clinic in November 2001. Retrospective data about diabetes, ischemic heart disease, Karnofsky score, hospitalization rate and causes of hospitalizations were collected. Hospitalizations of the year 2001 analyzed separately. The study included 111 patients. There were 123 hospitalizations in 75 patients, 1967 total hospital days. Admission rate was 1.34+/-1.5 per patient year, 19.82+/-25.96 hospital days per patient year, the mean length of each hospitalization--16.26+/-11.57. Most frequent causes of hospitalization--start of hemodialysis, infections, other causes, rare cause - hemodialysis complications. Karnofsky score correlated negatively with duration of hospitalizations per patient year. No statistically significant correlation was found between duration of hospitalizations and the presence of diabetes, ischemic heart disease, the way of starting hemodialysis (urgent or not). In comparison with other countries, in our center there were less hospitalizations but longer hospital days per admission. PMID- 12761432 TI - [A study of oxidative stress markers and antioxidant system activity in the serum of hemodialysis patients]. AB - The aim of this study was to evaluate lipid peroxidation status and anti-oxidant defenses in chronic hemodialysis patients comparison with healthy persons (control group). Forty-six hemodialysis patients were included in the study group and 66 healthy persons in control group. We determined the plasma levels of malondialdehyde as s biomarker of oxidative stress and the concentration of ceruloplasmin and activity catalase as antioxidant defenses. Our investigation indicate that the concentration of malondialdehyde increased 1.7 times in hemodialysis patients and activity of catalase decreased in 56.2% cases. Hyperlipidemia was evaluated in 20% of patients and hypoalbuminaemia in 6.6%. Most of these patients (48.8%) were found to have a history of occlusive vascular artery disease either coronary (17.7%), cerebral (11.1%) or peripheral (15.5%) occlusive disease. In conclusion these results suggest that intensity of lipid peroxidation and decreased plasma antioxidant potential in association with hyperlipidemia and hypoalbuminaemia may contribute to the increased risk of cardiovascular disease. PMID- 12761433 TI - [Relationship between lethality of hemodialysis patients, erythropoietin dosage for renal anemia treatment and hemodialysis quality]. AB - In December of 1999 and 2000 we visited all hemodialysis centers of Lithuania and collected data about all hemodialysis patients, using special questionnaires. The aim of the study was to evaluate the relationship between lethality of hemodialysis patients, erythropoietin dosage for renal anemia treatment and hemodialysis quality. The patients with higher Kt/V, higher levels of iron and albumin, normal levels of phosphorus and parathyroid hormone (PTH) requested lower doses of erythropoietin (analysis of the patients who were on hemodialysis in 2000 more than 6 months). So, we can conclude that adequate hemodialysis procedure and good management of hemodialysis patient are leading to the decrease request of erythropoietin doses for anemia treatment. We compared two groups of patients in order to examine relationship between hemodialysis quality and lethality of hemodialysis patients. We selected incident patients registered in December of 1999 and we divided these patients in December of 2000 in two groups: a) 175 patients, who continued hemodialysis treatment and b) 41 patients, who died in 2000. The results revealed, that dead patients were elder, their duration of weekly hemodialysis was shorter, Hb concentration lower, they had worse nutritional status (blood albumin level was lower). Lethality was associated with underlying diseases such as diabetes, hypertensive nephropathy and renal amyloidosis. PMID- 12761434 TI - [Structure of treatment expenditures for hemodialysis patients (data of Kaunas University of Medicine Hospital in the year 2001)]. AB - Prevalence of the end-stage renal disease patients is rising every year. In Lithuania the cost of hemodialysis procedure is under regulation of the Minister of Health. The objective of our study was to evaluate expenditures for hospitalizations and medical ambulatory treatment using data of Kaunas University of Medicine Hospital dialysis center in the year 2001. We analyzed ambulatory case records of all 111 end-stage renal disease patients dialyzed in our clinic in November 2001. Retrospective data about hospitalizations, hospital beds, expenditures for hospitalizations and medicamental ambulatory treatment were collected. There were 123 hospitalizations in 75 patients, 1967 total hospital days. Admission rate was 1.34+/-1.5 per patient year, 19.82+/-25.96 hospital beds per patient year, the mean duration of each hospitalization - 16.26+/-11.57 days. Expenditures for hospitalizations were 1415 litas ($ 354) per patient year. In average, hemodialysis procedures were carried out 2.5 times a week per patient (or 130 times a year) and expenditures for hemodialysis procedure were 36400 litas ($ 9100) per patient year. The mean number of medications monthly prescribed to hemodialyzed patients was 6.83+/-2.21. Expenditures for these medications were 16240 Lt ($ 4060) per patient year including expenditures for erythropoietin 14976 Lt per patient year. All health insurance expenditures for hemodialyzed patients were 54055 litas ($ 13514) and this was 2-4 fold lower than reported in developed countries. PMID- 12761435 TI - [Influence of hemodialysis on changes of trace metals concentrations in blood of patients with end-stage renal failure]. AB - This study on blood levels of trace metals in hemodialysis patients was carried out with an aim to clarify the impact of hemodialysis therapy on the trace metals status in such patients. Two hundred sixty six hemodialysis patients were enrolled. All patients underwent standard hemodialysis. Blood was collected before and after hemodialysis sessions. Atomic absorption spectrophotometry was applied to measure blood levels of aluminum, lead, cadmium, mercury as well as plasma levels of copper, and zinc. Aluminum, lead, mercury, and cadmium level in blood significant by increased after hemodialysis. The patients treated with acetate hemodialysis had significantly higher blood aluminum levels than the patients treated with bicarbonate hemodialysis (38.4+/-44.6 versus 25.9+/-43.5 microg/l, p=0.03). We observed negative correlation between copper concentration (n=60) in plasma and ultrafiltration (r=-0.4, p=0.003). The zinc and cadmium level increased with dialysis duration (r=0.2, p<0.001; r=0.2, p<0.02). Low plasma zinc levels, that is, <800 microg/l, was more often found in patients with <12 hour/week and <12 months hemodialysis duration. After 3 years of dialysis treatment blood lead, cadmium level (n=30) increased and plasma copper level decreased. PMID- 12761437 TI - [Control of viral hepatitis in Lithuanian hemodialysis centers in 1997-2001]. AB - Hemodialysis patients are a high-risk group for hepatitis B and C virus infections. The aim of the study was to evaluate the prevalence of B and C viral hepatitis, level of its control among patients on hemodialysis. In December of 1997, 1998, 1999, 2000, 2001 we visited all hemodialysis centers of Lithuania and gathered information about these infections in patients on hemodialysis. Eleven percent (11.4%) of all hemodialysis patients were not examined for HB(s)Ag and 15.2% for anti-HCV, 67.3%--for anti-HB(s) and 57.7% for anti-HBc in 2001. The number of examined patients for the markers of hepatitis had increased in 2001 in comparison with 1997. The same number of hemodialysis patients with HB(s)Ag was found in each year of study (14% in 1997, 14.4% in 2001). We observed the decrease in percentage of anti-HCV positive patients from 23% in 1998 till 15.4% (p<0.01) in 2001. Only 10.6% hepatitis B virus vaccinated patients was registered in 2001 and this percentage increased if compared to 6.3% in 1999. About (1/4) of anti-HB(c) positive patients were HB(s)Ag positive in 2000-2001. Chronic hepatitis B could be diagnosed for them. The duration of hemodialysis of HB(s)Ag and anti-HCV positive patients was longer compared to HB(s)Ag and anti-HCV negative patients (p<0.001). PMID- 12761436 TI - [Pathology of cardiovascular system among hemodialysis patients in Lithuania]. AB - During period of February to July 2002 the data of 602 hemodialysis patients were collected from 20 dialysis centres in Lithuania in order to determine the prevalence of cardiovascular diseases and their risk factors. In 34.9 percent of patients ischemic heart disease was diagnosed (myocardial infarction prior to hemodialysis, angina pectoris according to Rose's questionnaire, scar changes in ECG and/or hypokinesis zones in echocardiograms). Men were prevailing among the patients. The largest number of patients with ischaemic heart disease was on dialysis for one year, which proves that the patients start dialysis with the presence of pathology of cardiovascular system. Out of all patients, 45.3 percent were hypertensive. The majority of dialysis patients demonstrated risk factors for cardiovascular diseases: about 80 percent hypertrophy of the left heart ventricle, about a half of the patients were smokers, mainly men. Thirty-one percent of patients had overweight, 12.5 percent were obese. A half of the surveyed patients were not examined by means of echocardiography, one third did not undergo radiographic examination of thorax. It demonstrates insufficient attention to cardiovascular pathology as well as breach of the quality requirements for dialysis according to the order No 116 issued by the Ministry of Health March 15, 1999. PMID- 12761438 TI - [Influence of referral pattern to nephrologist on start of hemodialysis]. AB - In 2000-2001, 145 patients started hemodialysis due to end-stage renal disease in Hemodialysis unit at Kaunas University of Medicine. According to referral pattern to nephrologist all patients were divided into two groups: I group--< 3 months until start of hemodialysis--late referrals, II group--> 3 months until start of hemodialysis--early referrals. Comparing these groups, the results revealed worse clinical condition, much more catheterization of central vein (76.3%), prolonged hospitalization and major mortality within 12 moths from the start of hemodialysis, particularly within first 3 months, in late referrals. PMID- 12761439 TI - [Problems of arterio-venous access at the start of hemodialysis (results of Kaunas University of Medicine Hospital 1998-2002)]. AB - In 1998-2002 there were 93 patients, who started hemodialysis in Hemodialysis unit at Kaunas University of Medicine Hospital. According to start of hemodialysis all patients were divided into two groups: first group--patients (n=58, 62.3% ), who started hemodialysis through central venous catheter; second group--patients (n=35, 37.7%), who started hemodialysis on time through matured arterio-venous fistula. Comparing these groups, we noticed, that in the first group first hospitalization was longer, anemia was more severe and renal failure was more prominent, than in the second group. We analyzed the third group of patients (n=32), already on hemodialysis, which were hospitalized because of disfunction of arterio-venous fistula. The main cause of disfunction of arterio venous access were thrombosis and low blood flow. In urgent hemodialysis, punction of vena jugularis predominated vena subclavia punctions. More infection complications were noticed, in cases of vena subclavia punctions. PMID- 12761440 TI - [Nephrotoxicity of cyclosporin A after kidney transplantation]. AB - Cyclosporin A (CsA) is an effective immunosuppressive drug for the prophylaxis of rejection after organ transplantation. However, CsA is potentially toxic to various tissues: kidney, liver, pancreas, nervous system, etc. The aim of this study was to ascertain the frequency of CsA nephrotoxicity incidence according to the changes in graft biopsy material and its association with whole blood CsA levels. Data were obtained from 30 recipients after cadaver or living related kidney transplantation. All patients (pts) were divided into two groups: Gr1 included 17 pts with biopsy evidence of CsA damage and Gr2 -13 pts without these changes. The mean age of recipients (38.6+/-11.3 vs 34.6+/-13.3), donor and recipient human leucocyte antigen (HLA) match (2.5/6 vs 2.3/6), cold ischemic time (12.0+/- 9.3 h vs 13.8+/-10.3 h), percentage of kidney from cadaver donors (64.7% vs 69.2%) were similar in both groups. Comparison of CsA blood levels (>200 ng/ml) between Gr1 and Gr2 revealed statistically significant differences (70.6% vs 15.4%, p<0.05), correspondingly. The mean CsA blood level was higher in Gr1 (328.7+/-153.8 ng/ml vs 202.4+/-145.6 ng/ml, p<0.03). Thus, we suggest that CsA nephrotoxicity is associated with elevated CsA levels of more than 200 ng/ml. Biopsy is a very important criteria that helps to distinguish CsA nephrotoxicity and acute rejection. PMID- 12761441 TI - [Anti interleukin-2 receptor alpha antibodies (daclizumab) application for the treatment of kidney recipients]. AB - Acute rejection is a known risk factor for developing chronic allograft nephropathy, which remains as a major cause of long-term graft loss. Daclizumab (Zenapax), a humanized anti interleukin-2 receptor (IL-2R) alpha monoclonal antibody, is a novel selective immunosuppressive agent for the prophylaxis of acute rejection. Six patients (aged from 32 to 48) after cadaver and living donor kidney transplantation (Tx) have been treated with Daclizumab since 2000 in Lithuania. Daclizumab was prescribed (1.0 mg/kg intravenously before Tx and once every other week afterwards (five dozes in total) for all patients, except one, who received two dozes. The induction therapy was administered due to various immunological risk factors: retransplantated, sensitized and poorly HLA-matched patients, as well as non-immunological risk factors such as patients with diabetes, with kidney from sub-optimal donors, obesity of patients, etc. The maintenance immunosuppression consisted of cyclosporine, mycophenolate mofetil and prednisolon in all patients except one, who was treated without steroids. All the patients have been folloved-up for 5-36 months. Within the observation period in early time after Tx all the patients are alive and have no clinical signs of rejection. The graft function is normal (serum creatinine ranges between 100-120 micromol/l). CONCLUSION: Induction therapy with Daclizumab is safe and efficacious in preventing acute rejection in high risk kidney recipients. PMID- 12761442 TI - Effect of alpha-tocopherol on glucose uptake and contractility in rat skeletal muscle. AB - BACKGROUND: Free radicals and associated oxidative stress play an important role in the causation and subsequent complications of diabetes mellitus. Type I diabetes can be induced in experimental animals by administration of streptozotocin which causes selective destruction of B islet cells, probably by a free radical mediated mechanism. The objective of this study was to determine the effect of administration of alpha-tocopherol two weeks before and two weeks after streptozotocin injection on contractility and glucose uptake of rat hemidiaphragm. MATERIAL/METHODS: Male Wistar rats were used for this experiment. In the first phase, all the rats received an ordinary diet, while some were given alpha-tocopherol; in the second phase, diabetes was induced with streptozotocin, while the animals in the experimental group were given alpha-tocopherol two weeks before and two weeks after diabetes induction. RESULTS: Streptozotocin decreased the height of the indirect contraction and the time for 50% decrease in height of contraction of indirect responses. At the same time, the glucose uptake of the diaphragm muscle was also decreased. Administration of alpha-tocopherol (600 mg/kg-1, i.m.) ameliorated the inhibitory effect of diabetes on the skeletal muscle and reversed the decrease in time (s) of the 50% decrease in the height of contraction of the indirect responses of the rat hemidiaphragm preparation. Additionally, alpha-tocopherol increased glucose uptake by diaphragm muscles towards control level. CONCLUSIONS: Alpha-tocopherol supplementation probably protected against the inhibitory effect of oxidative stress observed in diabetes mellitus and contributed to improved contractility. PMID- 12761443 TI - Simvastatin reduces infarct size in a model of acute myocardial ischemia and reperfusion in the rat. AB - BACKGROUND: This study was designed to investigate the effects of simvastatin in a rat model of acute myocardial infarction. MATERIAL/METHODS: Male Wistar rats were anesthetized (thiopentone sodium 120 mg/kg). After a thoracotomy, the left anterior-descending coronary artery (LAD) was occluded (for 25 min) and reperfused (for 120 min). Area at risk (AR) was determined with Evans Blue dye and infarct size after staining of the area at risk with nitroblue tetrazolium. RESULTS: In rats, which received the vehicle for simvastatin (10% DMSO, 1 ml/kg i.v. at 1 h prior to the occlusion of the LAD), occlusion of the LAD for 25 min followed by reperfusion for 2 hours resulted in an infarct size of 54 +/- 4% (n=7) of the AR. When compared with vehicle, administration of simvastatin (1 mg/kg i.v. bolus administration at 1 h prior to the onset of myocardial ischemia) caused a significant reduction in myocardial infarct size of 39%. LAD-occlusion and reperfusion caused a progressive fall in mean arterial blood pressure (when compared with sham-operated animals). Simvastatin had no significant effect on blood pressure. CONCLUSIONS: The result indicates that simvastatin, an inhibitor of HMG-CoA reductase and a ligand of PPAR-a and PPAR-g, reduces the tissue necrosis associated with acute myocardial infarction. We propose that statins may be useful in conditions associated with ischemia-reperfusion of the heart and other organs. PMID- 12761444 TI - Protein kinase A expression and its possible roles in regulating tooth eruption genes in the dental follicle. AB - BACKGROUND: Tooth eruption requires the chronological expression of a series of genes in the dental follicle (DF). Protein kinase A (PKA) is a major phosphorylation pathway in the cells, and may regulate the expression of tooth eruption genes. MATERIAL/METHODS: In vivo, we studied the expression of the regulatory (R) and catalytic (C) subunits of PKA in the DF of newborn rats. In vitro, dental follicle cells (DFC) were treated with a specific PKA inhibitor, and then the gene expression of monocyte chemotactic protein-1 (MCP-1), colony stimulating factor-1 (CSF-1) and parathyroid hormone-related protein receptor (PTHrP-R) was determined. Cells also were treated with either phorbol-12 myristate-13-acetate or dibutyryl-cAMP, and the gene expression for RI alpha, RI beta, RII alpha and RII beta of PKA was examined. RESULTS: The results indicate that RI alpha of PKA is the predominant subunit in the DF with steady expression from days 1 to 11 postnatally. In contrast, expression of the RI beta, RII alpha, RII beta subunits are progressively reduced over this time period. However, there is a sharp decline of RI beta expression at postnatal day 3. The expression of the C subunits slightly decreases at days 3 and 5 with a greater decrease at day 7 postnatally. The specific PKA inhibitor reduces MCP-1 gene expression and translation, as well as moderately reducing CSF-1 and PTHrP-R expression. CONCLUSIONS: The reduction of the RI beta subunit in the rat DF at day 3 may result in an elevated PKA activity to trigger the maximal burst of gene expression of MCP-1 and CSF-1 seen at this time. PMID- 12761445 TI - High dose of caffeine administered to pregnant rats causes histopathological changes in the cornea of newborn pups. AB - BACKGROUND: Caffeine is frequently used during pregnancy and associated with teratogenic effects, such as low birth weight, hearth and digital defects, cleft palate and abortion with fetal loss. This study investigated histopathologically the effects of caffeine on neonatal rat cornea. MATERIAL/METHODS: Fifty pregnant Wistar-Albino rats (dams) were randomly divided into five groups, one control and four experimental. Between day 9 and 21 of gestation, group 1 dams (control, n=10) were exposed to intraperitoneal (i.p.) SF daily until delivery. Group 2 (n=10), group 3 (n=10) and group 4 (n=10) dams were treated with i.p. caffeine at doses of 25, 50 and 100 mg/kg/d, respectively, for the same period. Group 5 dams were given caffeine in distilled water in a gavage at a dose of 50 mg/kg/d during the same period. After normal delivery, the litters were killed at postnatal day 1 or 30 and the eyes were enucleated for corneal histopathologic investigation. RESULTS: Control and group 1 eyes had normal corneal epithelium, regular stromal fibers, descement membrane and monolayer inner corneal endothelium. The remaining experimental litters demonstrated changes, such as vacuolated endothelial cells with proliferation, hyperchromasia, polymorphism, endothelial cell agenesis, increased stromal mitotic activity and focal increase in corneal thickness with widely separated corneal lamellae in the injured area. These changes occurred most often in the litters treated with high doses of caffeine. CONCLUSIONS: Excessive gestational caffeine intake has been shown histopathologically to have some teratogenic effects on newborn rat cornea. PMID- 12761446 TI - Acetate metabolism in brain mechanisms of adaptation to ethanol. AB - BACKGROUND: The aim of this study was to estimate the role of acetate-induced metabolic changes in brain mechanisms of resistance to the narcotic effect of ethanol. MATERIAL/METHODS: Wistar rats were treated daily with ethanol (3.5 g/kg i.p. for 7 days). During alcohol treatment, the duration of ethanol-induced sleep was decreased. Levels of acetate, acetyl-CoA, adenosine, and the pool of AMP+ADP, and the activity of acetyl-CoA synthetase and 5'-nucleotidase in the brain were measured. Synaptosomal adenosine and acetylcholine release were measured in the presence of acetate, adenosine and 2-chloroadenosine. RESULTS: The concentration of acetate was higher in all investigated brain regions of ethanol rats in comparison to controls. The activity of acetyl-CoA synthetase and 5' nucleotidase, as well as the levels of adenosine and the pool of AMP+ADP were raised in cerebral cortex of ethanol rats. The synaptosomal level of adenosine was higher in control rats. Acetate caused a 3-fold increase of extrasynaptosomal adenosine level. Synaptosomes from ethanol rats showed higher rates of Ca dependent releases of acetylcholine. 2-chloradenosine resulted in inhibition of synaptic acetylcholine release only in control rats. CONCLUSIONS: Ethanol causes an increase of acetate in brain and adenosine level in cerebral cortex. Acetate causes an increase of the extrasynaptic adenosine level. Prolonged ethanol treatment results in an increase of synaptic Ca-dependent acetylcholine release. Seven-day treatment with ethanol eliminates the inhibitory effect of 2 chloroadenosine-A1 adenosine receptor agonist on synaptic Ca-dependent acetylcholine release. PMID- 12761447 TI - Magnesium homeostasis disorders caused by experimental acute pancreatitis. AB - BACKGROUND: In this study we set out to examine changes in magnesium concentration in serum and tissues of rat and the direction of its shifts in tissue concentration in the course of experimental pancreatitis. MATERIAL/METHODS: The experiment was performed on 200 male Wistar rats, weight 250-350 g, according to Heinkel and Aho's method. Four groups of rats were randomly selected: Z-healthy (20), K1-first control (60), K2-second control (60), D-experimental (60). In Group D acute pancreatitis was induced by injection under anesthesia; in Group K2, physiological fluid was injected; in Group K1, only the needle was inserted. At the 6th, 12th, 24th and 48th experimental hours blood and some organs were collected from 15 animals in each group. RESULTS: The magnesium level in serum and in the organs was within the normal range for the K1 rats, not significantly different from controls. The lowest serum and kidney concentrations of magnesium were observed after 6 and 12 hours in groups K2 and D. The lowest magnesium concentration in pancreas was observed after 12 and 24 hours in the same two groups. The highest magnesium concentration in liver was detected in the K2 and D animals after 6 and 12 hours, and in heart after 12 hours. Similar changes in the serum glucose level were observed in groups K2 and D. CONCLUSIONS: Our results demonstrate that in the course of experimental acute pancreatitis there is a decreased magnesium concentration in blood serum, pancreas and kidney. PMID- 12761448 TI - Genotyping of p53 codon 175 in colorectal cancer. AB - BACKGROUND: The gene encoding the tumor suppressor protein p53 is one of the most frequently mutated in human cancer. Over 8000 mutations of this gene have been identified. The spectrum of p53 mutations varies among tumor types. More than half of these mutations appear in three hotspot codons: 175, 248 and 273. The purpose of the present study was to evaluate the GgA transition in codon 175 of the p53 gene as a potential marker of colon cancer progression. MATERIAL/METHODS: The prognostic significance of alterations in codon 175 exon 5 of p53 gene was studied in 32 patients with colorectal cancer. The ASA-PCR method was used to assess GgA (ArggHis) transition in codon 175. RESULTS: The analyzed point mutation was identified in 56% of the tested cases of colorectal cancer. The frequency of codon 175 p53 gene mutation was found to increase with cancer aggressiveness. Screening for the studied point mutation in codon 175 showed no alteration in analyzed samples classified as Dukes' A stage, while variations were observed in 43%, 50% and 75% of the cases classified as Dukes' B, C, and D, respectively. In three cases the studied mutation was also found in some mucosal cells obtained from resectional borders. CONCLUSIONS: The GgA transition in codon 175 of the p53 gene may be useful as a potential marker of colorectal cancer progression and as a means of evaluating the margins of surgical resection. PMID- 12761449 TI - Utility of the neurologic intensive care evaluation (NICE) in detecting neurologic deficit after cardiac operations--a pilot study. AB - BACKGROUND: Neurocognitive changes are common after cardiac operations. The acute post-operative period is a critical time when significant neurologic changes may be detected and appropriate therapy initiated promptly. Formal neuropsychologic testing in this situation however is impractical so other means of early detection are required. The goal of this preliminary study was to determine whether simple, standardized, serial nursing neurologic evaluations using the Neurologic Intensive Care Evaluation (NICE) could be helpful in screening patients for neurologic injury in the immediate post-operative period. MATERIAL/METHODS: Details of the intra-operative and post-operative anesthetic management were obtained during report and nurses subsequently scored acute post operative patients in the CTSICU using the NICE every half hour for the first five hours. Finally, a chart review was performed to determine the neurologic outcome of the patients. RESULTS: The time to achieve the lower NICE scores which reflect mainly brainstem function was the same whereas the time to achieve NICE scores>4 was prolonged in patients with neurocognitive dysfunction. The effect of intra-operative factors on the times to achieve NICE scores was different for the higher and lower scores. The times to reach NICE scores correlated with outcome variables including the time in ICU and time of intubation. CONCLUSIONS: Standardized, serial nursing neurologic assessments of post-operative cardiac patients may be a useful tool for early identification of patients with neurologic injury. They may also provide useful information complementing the data obtained from detailed neuropsychologic testing on the neurologic effects of cardiac operations. PMID- 12761450 TI - Adenosine-activated myocardial ischemia does not induce delayed preconditioning. AB - BACKGROUND: Our objective was to determine whether adenosine-induced ischemia exerts a delayed cardiac protective effect in patients with stable effort angina ischemic heart disease. MATERIAL/METHODS: The study group was comprised of 32 patients (men) with symptoms of stable effort angina, aged 38-65 years (Group 1), and 18 clinically healthy subjects (3 women, 15 men), aged 35-55 years (Control group). The study protocol included baseline ECG and treadmill echocardiogram (ET1); ECG and adenosine echocardiogram performed 7 days after ET1; repeated exercise test exactly 24 h after adenosine infusion (ET2). Increases in heart rate (HR), systolic (SBP) and diastolic (DBP) blood pressure, maximum ST-segment depression (max IST) and total ST-segment depression (SIST) on ECG were compared, as well as left ventricular end-diastolic volume (LVEDV), end-systolic (LVESV) volume, ejection fraction (EF), and wall motion synergy index (WMSI). RESULTS: No statistically significant differences were found in the increased values of the investigated electrocardiographic and echocardiographic parameters in either group on either exercise test. The only positive trend was observed in LVEDV. In Group 1 LVEDV increased significantly from rest values during ET1, whereas during ET2 LVEDV did not change. CONCLUSIONS: Adenosine-induced ischemia does not exert a delayed protective effect in respect to cardiac bioelectrical and mechanical functions in patients with ischemic heart disease in the form of stable effort angina. PMID- 12761451 TI - Serum free testosterone in men with coronary artery atherosclerosis. AB - BACKGROUND: The role of testosterone in the pathogenesis of atherosclerotic changes has not been fully explained. The aim of our work was to determine whether levels of serum free testosterone differ between men with and without coronary artery disease. MATERIAL/METHODS: We examined the serum concentration of free testosterone by the radioimmunoassay method in two groups of subjects, carefully matched for body mass index and age: 105 men with angiographically diagnosed coronary artery disease, and 23 control subjects without any changes in coronary arteries. We also evaluated the concentrations of total cholesterol, HDL cholesterol, LDL-cholesterol and triglycerides by enzymatic methods. RESULTS: Men with coronary artery disease had significantly lower levels of free testosterone than did controls. The concentration of free testosterone diminished with age, and a negative correlation was found between free testosterone and body mass index. We did not discover any connection between the concentration of free testosterone and lipids. CONCLUSIONS: A low level of free testosterone may be related to the development of coronary artery disease. PMID- 12761452 TI - Familial dilated cardiomyopathy: evidence for clinical and immunogenetic heterogeneity. AB - BACKGROUND: There is increasing awareness of the familial nature of dilated cardiomyopathy (DCM). Mutations in the genes coding for cytoskeletal and sarcomere proteins have been identified. Phenotyping of familial DCM (FDCM) may help to improve genetic diagnosis. The aim of our study was to evaluate the clinical features, pattern of transmission, and immunogenetic data of FDCM. MATERIAL/METHODS: We obtained family histories in order to construct pedigrees and prospectively evaluated 204 family members of 27 patients with angiographically proven DCM. FDCM was defined as more than 1 person with DCM in a family. The study protocol included repeated clinical examination, electrocardiography, echocardiography and blood sampling. RESULTS: Among the families, we identified the following phenotypes: DCM with conduction defects (n=2), early onset DCM with a rapid course in male relatives (n=2), and DCM preceded by ventricular arrhythmia (n=1). The remaining families presented with a heterogeneous course of the disease. The disease was transmitted in an autosomal dominant fashion in 14 of our pedigrees, possibly X-linked in three and indeterminate in 10 sib-pairs. The frequency of the DRB1*04 allele was low in probands with the disease (3/20, 15%); heterozygozity for DRB1*03/DRB1*04, known to increase susceptibility to IDDM1, was identified in 2 of 20 DCM probands (10%). CONCLUSIONS: Familial dilated cardiomyopathy is a heterogeneous disorder; autosomal dominant transmission is most common. The distinct clinical phenotypes and specific immunogenetic features found in some families indicate that different pathogenetic mechanisms can lead to the PMID- 12761453 TI - Differentiating tuberculous from malignant pleural effusions: a scoring model. AB - BACKGROUND: Patients with tuberculous or malignant pleural effusions frequently have similar clinical manifestations and pleural fluid profile. The aim of our study was to derive a simple clinical score for differential diagnosis of these two clinical entities. MATERIAL/METHODS: Our retrospective study involved 106 patients with tuberculous pleurisy and 286 with malignant effusions, seen during a 9-year period. Clinical and laboratory variables with (model 1) and without (model 2) the addition of pleural adenosine deaminase entered into a multivariate analysis to calculate a scoring system (range 0 to 10) for the detection of tuberculous effusions. RESULTS: In model 1, four variables predicted a tuberculous etiology: adenosine deaminase > or = 40 U/L (5 points), age <35 years (2), temperature > or 37.8 degrees C (2), and pleural fluid red blood cell count < 5 x 10(9)/L (1). In addition to the last three items, model 2 identified other predictive parameters: no history of malignancy (3), pleural protein > or = 50 g/L (1), and pleural fluid to serum lactate dehydrogenase ratio > or 2.2 (1). Summated scores of > or 5 in model 1 and > or 6 in model 2 yielded measures of sensitivity (95% and 97%), and specificity (94% and 91%) for discriminating tuberculous from malignant effusions, respectively. The area under the ROC curve for models 1 and 2 was 0.987 and 0.982, respectively. CONCLUSIONS: The combination of clinical data and pleural fluid chemistry profile into a score based model can facilitate differential diagnosis between tuberculous and malignant effusions. PMID- 12761454 TI - Selected factors associated with diabetes mellitus in a rural Palestinian community. AB - BACKGROUND: The purpose of this study was to investigate the distribution and association of selected risk factors for diabetes mellitus in a semi-rural Palestinian village. MATERIAL/METHODS: We performed a cross-sectional, population based study of 500 adults aged 30 to 65 in a semi-rural Palestinian village. The study included two phases: a household survey and an individual assessment utilizing the oral glucose tolerance test OGTT to determine the diabetes status of the participants, anthropometric measurements for body mass index (BMI) and waist-hip ratio (WHR), blood biochemistry measurements for lipids, blood pressure measurement, and a standard questionnaire to assess demographic and other factors. RESULTS: The association between various risk factors and diabetes status was explored by comparing statistical means and proportions and crude and adjusted odds ratios (OR). A multivariate logistic regression using sex and seven factors initially found to be significantly associated with diabetes identified four factors that remained significantly associated with diabetes after adjustment for age and sex. The four main factors are age (OR = 1.08, 95%CI = 1.05-1.12), positive family history of diabetes (OR = 3.09, 95%CI = 1.53-6.24), triglycerides (OR = 1.006, 95%CI = 1.002-1.009), and WHR (OR = 2.13, 95%CI =1.31 3.45). CONCLUSIONS: Two factors associated with diabetes are potentially modifiable in this Palestinian population: WHR and triglyceride levels. These should be addressed by preventive health activities, including health promotion. The association between diabetes and age supports the conclusion that the prevalence of diabetes is expected to increase with the aging of the population. PMID- 12761455 TI - Insulin resistance in the first-degree relatives of persons with type 2 diabetes. AB - BACKGROUND: Type 2 diabetes is characterized by impaired insulin secretion and insulin resistance. Both genetic and environmental factors are thought to play a role in the pathogenesis of this disease. Obesity is associated with increased risk of type 2 diabetes, but some metabolic abnormalities may also be present in lean, predisposed subjects. The aim of the present study was to assess insulin sensitivity in lean normoglycemic offspring of parents with type 2 diabetes. MATERIAL/METHODS: We examined 17 lean offspring (BMI<25 kg x m-2) whose parents had type 2 diabetes, and 17 age, sex and BMI-matched subjects without family history of diabetes as controls. Anthropometric and biochemical measurements, oral glucose tolerance test (OGTT), and euglycemic hyperinsulinemic clamp were performed. RESULTS: The subjects did not differ in anthropometric measurements, and all had normal glucose tolerance according to WHO criteria. Offspring of type 2 diabetic parents were markedly more hyperinsulinemic (p<0.05) and insulin resistant (p<0.005). The ratio of the increase in insulin to the increase in glucose during the first 30 minutes of the OGTT, a crude index of first-phase insulin secretion, did not differ between groups. Insulin sensitivity was negatively related to fasting plasma insulin (r=-0.67, p<0.001) and non esterified fatty acids (r=-0.43, p<0.02). CONCLUSIONS: Insulin resistance is present even in young lean subjects at high risk to develop type 2 diabetes. Our data suggest that insulin resistance may be a primary abnormality in the pathogenesis of this disease. PMID- 12761456 TI - Colonic pacing in patients with constipation due to colonic inertia. AB - BACKGROUND: The colon exhibits electrical activity in the form of pacesetter potentials (PPs) and action potentials (APs); no activity is recorded in total colonic inertia (TCI). Electrical activity mediates colonic motility. We have elsewhere demonstrated that 4 pacemakers initiate colonic electric activity. We investigated the hypothesis that the absence of electric activity in TCI is due to non-functioning pacemakers and that their stimulation produces colonic motility and defecation. MATERIAL/METHODS: 19 patients with TCI were divided into 2 groups: a study group of 11 patients (age 42.6 +/- 6.8 years; 7 women) and a control group of 8 patients (age 41.4 +/- 6.3 years, 5 women). 7 healthy volunteers (age 40.9 +/- 6.1 years, 5 women) were also studied. Pacing electrodes were endoscopically hooked to colonic mucosa: one stimulating and 2 or 3 recording. Stimulating electrodes were applied to the 4 potential pacemaker sites located at the cecal pole, the cecocolonic junction, the mid-transverse colon, and the colosigmoid junction. Electrical activity was recorded before and during electrical stimulation. RESULTS: Colonic pacing of healthy volunteers caused a significant increase in the frequency, amplitude and conduction velocity of basal electric waves. Colonic pacing in the study group produced PPs and APs, which varied between colonic segments. No waves were recorded from the control group in which the pacemakers were not activated. CONCLUSIONS: Colonic pacing produces electrical activity in patients with TCI. This method can be applied for the treatment of constipation in these patients. PMID- 12761457 TI - Occupational exposure to HIV infection in health care workers. AB - BACKGROUND: The aim of this study was to determine the epidemiology of occupational exposure to HIV infection in health care workers (HCWs). MATERIAL/METHODS: A survey from 4 hospitals between February 1995 and May 2001 identified 28 HCWs who had been exposed to HIV. The type of exposure to HIV, the circumstances of the incident, the safety precautions applied, and epidemiological information were evaluated in each case. A blood specimen for HIV serological testing was collected at the baseline visit and at 6 weeks, 3 months, 6 months and 12 months follow-up. RESULTS: The study population consisted of 24 women and 4 men, mean age 34.7 +/- 5.8, including 15 nurses, 9 physicians, 2 nursing assistants, 1 morgue worker, and 1 medical student. These workers had been exposed to blood and infectious body fluid from patients who had AIDS (17 exposures), were HIV-antibody positive and symptomatic (3 exposures), or were HIV antibody positive and asymptomatic (8 exposures). The exposure types included percutaneous injury (22) and blood or infectious body fluid contact with mucous membranes (2) or intact skin (4). Post-exposure chemoprophylaxis was used by 18 HCWs, 12 of whom reported side effects. None of the HCWs was HIV-seropositive in follow-up after occupational exposure. CONCLUSIONS: Nurses are most at risk for occupational exposure to HIV infection. Post-exposure chemoprophylaxis is effective. Routine post-exposure management is also a good instrument to detect serological markers of HBV and HCV infection among HCWs and patients. PMID- 12761458 TI - Use of drotrecorgin alfa--recombinant activated human protein C in treatment of septic shock in the course of therapy for nephrolithiasis. AB - BACKGROUND: Septic shock is the most dangerous complication of nephrolithiasis management utilizing percutaneous methods. CASE REPORT: The patient, D.M., aged 60 was subjected to scheduled percutaneous nephrolithotomy due to coral calculosis of the pyelocalyceal system. As pyuria was noted intraoperatively, nephrostomy was left after the procedure. Over ten hours after the surgery the patient developed the symptoms of severe septic shock with progressive respiratory distress, renal failure, intravascular coagulation syndrome and impaired consciousness. Nephrectomy was performed, antibiotic treatment and high doses of norepinephrine instituted. Hemodynamic stabilization was obtained, without, however, marked improvement of the patient's condition. Because of persistent coagulation disorders and multiorgan dysfunction, recombinant activated protein C preparation--drotrecorgin alfa (Xigris Eli Lilly) was added to the therapeutic regimen. From the second day of infusion, systematic improvement of the coagulation system parameters was observed, making it possible to reduce the doses of catecholamines, oxygen concentration in the gas mixture used for ventilation, as well as stabilization of the function of the preserved kidney. The patient was weaned off the respirator on the 8th day of treatment and on the 13th day referred to the Urology Department, from which she was soon discharged home. CONCLUSIONS: 1. Surgical resection of the infection source and cause of the septic shock is the prerequisite for successful pharmacological treatment. 2. Administration of rh-APC to a patient meeting the PROWESS criteria may be an effective and safe method of treatment in the complex management of septic shock. PMID- 12761459 TI - Absent end diastolic flow in umbilical artery and umbilical cord thrombosis at term of pregnancy. AB - BACKGROUND: Meconium staining of the fetus and placenta is associated with increased neonatal mortality and asphyxia. Very often it is unclear whether the discharge of meconium is a cause or an effect of fetal distress. In the available literature there are no large epidemiological studies of pregnancy outcome with meconium-related lesions, even though this could be useful to improve our state of knowledge on this topic. CASE REPORT: A case of umbilical cord vascular necrosis is described. A severely asphyxiated infant was delivered at 39 weeks' gestation by cesarean section due to alarming results of fetal heart rate monitoring and rupture of membranes with meconium-stained amniotic fluid. There was no meconium aspiration. We report a review of 15 similar cases. In the whole series, a linkage between umbilical cord vascular necrosis and evidence of remote meconium discharge always seems to be detectable. The pathophysiological mechanism is unknown. CONCLUSIONS: It is still not clear why only a tiny percentage of cases with meconium-stained amniotic fluid develops umbilical cord lesions and poor pregnancy outcome. Further investigations are needed to explain why some meconium-stained newborns suffer severe neurological and other damage even without meconium aspiration. PMID- 12761460 TI - Respiratory failure due to bilateral diaphragm palsy as an early manifestation of ALS. AB - BACKGROUND: Diaphragm palsy is common in the advanced stages of motor neuron disease and is a primary cause of fatal outcome However, respiratory failure is a presenting symptom of motor neuron disease in only a small number of patients. CASE REPORT: We present the case of a patient with dyspnea and orthopnea followed by subacute respiratory failure due to bilateral diaphragm paralysis as the first manifestation of amyotrophic lateral sclerosis. CONCLUSIONS: Failure of respiratory muscle function can be the first symptom of motor neuron disease, and may precede clinical manifestation in voluntary motor units in ALS. Therefore, in cases of unexplained acute respiratory failure or when respiratory support must be continued for no clear reason, a motor neuron disease such as amyotrophic lateral sclerosis, which leads to respiratory muscle weakness and diaphragm paralysis, should be taken into consideration. PMID- 12761461 TI - Feather duvet lung. AB - BACKGROUND: Bird fancier's lung (BFL) is a type of hypersensitivity pneumonitis, which is induced by inhalation of bird related antigens. The diagnosis of BFL induced by feathers is difficult because feathers are generally not recognized as a causative antigen of BFL. We report a female case of chronic BFL presumably due to a feather duvet, which presents as pulmonary fibrosis. CASE REPORT: A 73 year old woman presented with exertional dyspnea for the last three years. She had raised two pigeons for three years (1971-1973) in her forties and had been using a feather duvet for the last eight years (1992-2000). A chest X-ray showed reticular infiltrates in the both peripheral lung field and an HRCT scan showed scattered consolidation, micronodules, and peribronchial ground-glass opacities. Lymphocyte proliferation to the feather antigen was positive and inhalation provocation test using a bird antigen was also positive. Thoracoscopic biopsy specimens showed organization, cholesterol clefts, alveolitis around terminal and respiratory bronchioles--all of which are consistent with chronic hypersensitivity pneumonitis. Clinical findings have spontaneously improved after she stopped using her feather duvet. CONCLUSIONS: Feather beds including duvets, pillows, and cushions are now popular all over the world. Physicians should be aware of feathers as a cause of BFL since this induction seems to be more prevalent. PMID- 12761462 TI - Concomitant heterozygous factor V Leiden mutation and homozygous prothrombin gene variant (G20210A) in patient with cerebral venous thrombosis. AB - BACKGROUND: Factor V Leiden mutation represents the most common genetic risk factor of venous thrombosis in Caucasian population. A common mutation in prothrombin gene, which is due to G-->A transition at position 20210, is also associated with elevated prothrombin concentration and thrombosis. Both this mutations may constitute concomitant risk factors for deep venous thrombosis. CASE REPORT: A 29-years old woman was admitted in the Emergency Department because of severe headache with vomiting that she suffered for a few days without any neurological deficits. In the Emergency she presented first in her life tonic clonic seizures followed by right hemiparesis and aphasia and than was admitted to hospital. CT and MR scan showed a large lesion in the left fronto-parietal region with extent edema, which was first diagnosed as tumor. Following MR showed more lesions and typical signs of sinus thrombosis. She improved quickly after stroke without any anticoagulant treatment. Genetic study revealed factor V Leiden mutation and homozygous mutation G20210A in prothrombin gene. CONCLUSIONS: Both mutations found in this case, alone, are not a high risk factors for venous thrombosis but together may increase 5-10 fold risk of venous thrombosis. Venous stroke must be considered always in acute neurological events with organic brain lesions, especially in young PMID- 12761464 TI - Health, 'small-worlds', fractals and complex networks: an emerging field. AB - The importance of 'small-worlds', fractals and complex networks to medicine are discussed. The interrelationship between the concepts is highlighted. 'Small worlds'--where large populations are linked at the level of the individual have considerable importance for understanding disease transmission. Complex networks where linkages are based on the concept 'the rich get richer' are fundamental in the medical sciences--from enzymatic interactions at the subcellular level to social interactions such as sexual liaisons. Mathematically 'the rich get richer' can be modeled as a power law. Fractal architecture and time sequences can also be modeled by power laws and are ubiquitous in nature with many important examples in medicine. The potential of fractal life support--the return of physiological time sequences to devices such as mechanical ventilators and cardiopulmonary bypass pumps--is presented in the context of a failing complex network. Experimental work suggests that using fractal time sequences improves support of failing organs. Medicine, as a science has much to gain by embracing the interrelated concepts of 'small-worlds', fractals and complex networks. By so doing, medicine will move from the historical reductionist approach toward a more holistic one. PMID- 12761463 TI - Induction of oral tolerance as treatment or prevention of chronic diseases associated with Chlamydia pneumoniae infection--hypothesis. AB - BACKGROUND: Chlamydia pneumoniae infection is associated with chronic diseases such as asthma, reactive arthritis, and atherosclerosis. Several investigations and experimental results indicate an excessive immune response to heat shock protein (hsp) 60 as a possible common pathogenetic link between Chlamydia pneumoniae infection and the associated chronic diseases. CASE REPORT: A 46-year old woman with persistent C. pneumoniae infection and Reiter's syndrome had been treated for three years with antibiotics and nonsteroidal anti-inflammatory drugs without success. However, she was repeatedly free of complaints for several months following two-week cycles taking oral dilutions of Chlamydia trachomatis daily. CONCLUSIONS: The placebo effect does not seem sufficient to explain the clinical benefits obtained repeatedly by drinking dilutions of C. trachomatis. Induction of oral tolerance (immune modulation) to hsp60 may have occurred, leading to the clinical benefits. The known risk factors for atherosclerosis do not account for all cases of atherosclerosis. If autoimmunity to hsp60 is involved in the pathogenesis of atherosclerosis, so natural acquisition of oral tolerance to hsp60 may contribute to the geographical differences in atherosclerosis prevalence. PMID- 12761466 TI - Measurement and application of arterial stiffness in clinical research: focus on new methodologies and diabetes mellitus. AB - The excess risk of cardiovascular disease in patients with Type 2 diabetes mellitus remains largely unexplained. Arterial stiffness, an early feature of diabetic vasculopathy involving several mechanisms, results in impaired arterial compliance, and has recently been proposed as a powerful independent predictor of cardiovascular disease. Increased arterial stiffness can contribute to the development and progression of hypertension, left ventricular hypertrophy and dysfunction, and to decreased myocardial perfusion, all of which are highly prevalent in Type 2 diabetes mellitus. Large artery stiffening has been demonstrated in Type 2 diabetes using several different methods including measurement of central pulse wave velocity, or estimation of aortic compliance, a technically demanding technique requiring the simultaneous measurement of stroke volume and diastolic pressure decay. Increased arterial stiffness of smaller arteries is also an important contributor to systemic arterial stiffness and cardiovascular risk. Increasing demand by clinical researchers for measurement of arterial stiffness has led to the development and commercial availability of highly practicable techniques. These new techniques, which utilise different aspects of the pulse pressure waveform, are simple, reliable and reflect both large and small vessel stiffness. They offer new tools for identifying patients at increased risk of developing cardiovascular complications including those with established diabetes. In this article we introduce the reader to the concepts of arterial stiffness, the significance of arterial stiffness for diabetes, and the new techniques that may potentially be useful for clinical researchers or practitioners. PMID- 12761465 TI - Methoxychlor modifies the ultradian secretory pattern of prolactin and affects its TRH response. AB - BACKGROUND: Methoxychlor (MTX) is an insecticide characterized by both estrogenic and antiandrogenic properties that may modify the secretory pattern of prolactin. The objective of the present work was to analyze MTX's effects on changes in the episodic secretion of prolactin induced by thyrotropin-releasing hormone (TRH). MATERIAL/METHODS: Adult male Sprague-Dawley rats (400-420 g body weight) were administered 25 or 50 mg/kg/day of MTX s.c. in sesame oil for 30 days. Control animals received the vehicle only. RESULTS: In animals treated with 50 mg/kg/day of MTX, mean serum prolactin levels and absolute pulse amplitude increased (p< or = 0.05 and p< or = 0.001 respectively). TRH administration (two pulses of 2 Kg) increased both mean serum prolactin levels and the absolute amplitude of the prolactin peaks in all groups studied (p< or = 0.001). The response to TRH was greatly increased in the animals treated with 50 mg/kg/day. TRH reduced the mean life of the hormone in control rats (p< or = 0.01) and in animals treated with 50 mg/kg/day (p< or = 0.001). After TRH administration, the duration of prolactin peaks was increased in the MTX-treated animals (p< or = 0.05 and p< or = 0.001 for 25 and 50 mg/kg/day respectively). TRH reduced the frequency of prolactin pulses in the animals treated with 50 mg/kg/day (p< or = 0.05). CONCLUSIONS: These data suggest that MTX sensitizes the pituitary to TRH, indicating a direct effect of MTX on the pituitary gland. PMID- 12761467 TI - Decision tree and paradigms of primary breast cancer: changes elicited by preoperative therapy. AB - A small difference in decision-making causes a big impact on the long-term outcome in cancer treatment. Novel methods such as preoperative systemic treatment or sentinel node biopsy (SNB) may alter the decision-trees in primary breast cancer management. Recent data showed that preoperative chemotherapy drives pathological complete response (pCR) that implies long-term relapse-free, for about 25% to 30% of early breast cancer patients. In fact, preoperative systemic therapy has come to be used regardless of the tumor stage. From the point of the clinical trial, pCR can be recognized as a new endpoint, which promises to speed up new therapy development. Therefore, it is crucial to consider new paradigms including preoperative therapy in the decision-tree. There are several criticisms regarding the preoperative therapy, such as a possibility of over-treatment, however these issues might be resolved by changing the concept or procedures slightly. For instance, if SNB is conducted before the treatment, the over-treatment issue can be eliminated. In this article, we will discuss the changes in decision-tree and paradigms for primary PMID- 12761468 TI - Sound therapy induced relaxation: down regulating stress processes and pathologies. AB - The use of music as a means of inducing positive emotions and subsequent relaxation has been studied extensively by researchers. A great deal of this research has centered on the use of music as a means of reducing feelings of anxiety and stress as well as aiding in the relief of numerous pathologies. The precise mechanism responsible for these mediated effects has never been truly determined. In the current report we propose that nitric oxide (NO) is the molecule chiefly responsible for these physiological and psychological relaxing effects. Furthermore this molecules importance extends beyond the mechanistic, and is required for the development of the very process that it mediates. Nitric oxide has been determined to aid in the development of the auditory system and participate in cochlear blood flow. We show that NO is additionally responsible for the induced exhibited physiological effects. We proceed to outline the precise neurochemical pathway leading to these effects. Furthermore we explore the interrelationship between the varying emotion centers within the central nervous system and explain how the introduction of music can mediate its effects via NO coupled to these complex pathways. PMID- 12761469 TI - Increased clinical correlation in anatomy teaching enhances students' performance in the course and National Board subject examination. AB - The purpose of this study is to determine the effectiveness of increased clinical correlation teaching in a first year anatomy program and subsequent student performance on departmental and nationally standardized examinations. Basic science curricula in medical schools are increasingly being taught with clinical correlation in order to provide a >seamless transition' from preclinical-to clinical years. The National Board examination for basic science is also increasingly clinically oriented and designed to test students problem solving and applied skills. Five-year period data of students course grades and standardized subject examination scores are compared statistically in this study. The results indicate that incorporation of clinical correlation teaching and more problem-focused assessment of student learning resulted in better performance on the standardized National Board anatomy examination. We suggest that such an approach can be easily adopted in other preclinical courses. PMID- 12761471 TI - The role of hypoxia and nitrogen monoxide in the regulation of cellular iron metabolism. PMID- 12761472 TI - Research ethics committees in developing countries and informed consent: with special reference to Turkey. AB - Research ethics committees (RECs) are the institutional equivalent of institutional review boards (IRBs) with respect to their role in the ethical analysis of research projects. In Turkey, as in most developing countries, these committees were initiated as a result of pressure from the Western scientific community. Since their formation, RECs have faced problems because of an underdeveloped scientific culture and the absence of established ethical standards. On the one hand, the standards of the RECs and IRBs in the developed world seemed too hard to achieve; on the other, some of the international regulatory rules, such as respect for autonomy, are not culturally sensitive. In addition to problems facing RECs, researchers have had difficulties in fulfilling RECs' requisites. Respect for autonomy is a good example for Turkey. The social construct of Turkish society is not based on the Western concept of autonomy; it is based on "collective autonomy," which is completely different from the Western definition. Cultural interpretation of concepts is the perception of concepts effected by their cultural connections with other concepts. For example, "motherhood" as a concept has many cultural connections, so when it is used we have to consider those culturally determined contexts. In some countries, the familiar concept and term are kept separated from the one that is imported. In others, the imported concept alters the original concepts content. Although its course varies in different countries, it always puts extraordinary pressure on RECs and researchers in their relationship between themselves and other parties with conflicting interests. In this article, some of the main concerns of RECs in developing countries are discussed, with special reference to Turkey. The universality of regulatory norms is questioned and the importance of culturally sensitive and effective safeguards emphasized. PMID- 12761473 TI - Differential gene expression in uterine leiomyoma. AB - Uterine leiomyomas are the most common tumor of the genitourinary system in women and are a major cause of morbidity. The molecular causes of the disease remain unclear. In this study, we examined gene expression in leiomyomas and normal myometrium. RNA was prepared and gene expression determined with the use of Affymetrix GeneChip U_95 arrays containing approximately 12,000 known genes and 48,000 expression sequence tags. Several genes were found to be differentially expressed in these two sample sets, and these genes were analyzed for their expression in a variety of other normal and diseased tissues. Four genes- doublecortin, calpain 6, interleukin-17B, and proteolipid protein 1--were found to be overexpressed in leiomyomas compared with normal myometrium and eighteen other tissues. Sets of genes were identified whose expression could be used to cluster samples with leiomyomas or normal myometrium with the use of Eisen Cluster software. We conclude that differences in gene expression can be detected between leiomyomas and normal myometrium and that these changes in gene expression may yield clues to the pathophysiology of this common tumor. PMID- 12761474 TI - Effects of hypoxia and nitric oxide on ferritin content of alveolar cells. AB - Concentrations of ferritin in alveolar cells and on the alveolar surface are increased in patients with a variety of respiratory disorders. Ferritin synthesis by cells is modulated by iron content but is also influenced by stimuli other than iron. In this study we sought to determine whether in vitro exposure to hypoxia- or nitric oxide (NO)-induced ferritin accumulation or release by human alveolar macrophages (AMs) or a lung cancer-derived epithelial cell line (A549). Changes in cell content of iron and ferritin (L- and H-types), as well as ferritin content of cell supernatants, were determined after in vitro exposure to hypoxia (1% or 10% O(2), 18 hours) or the NO donor S-nitroso-N acetylpenicillamine (SNAP, 0.01-1.0 mmol/L, 18 hours). Exposure to 1% O(2) increased ferritin content in both cell types (>fourfold increase; P <.005) without changing iron content. Treatment with SNAP increased ferritin content of A549 cells in a dose-dependent manner, whereas treatment of AMs decreased cellular iron and ferritin content and increased supernate ferritin content. Pretreatment of cells with N-acetylcysteine (500 micromol/L) reduced hypoxia induced ferritin accumulation in alveolar cells and completely inhibited NO induced ferritin accumulation in A549 cells. These findings indicate that exposure to 1% O(2)can increase ferritin content in alveolar cells, whereas NO can increase ferritin content (A549 cells) or decrease ferritin content (AMs). PMID- 12761475 TI - Differential effects of low-dose docosahexaenoic acid and eicosapentaenoic acid on the regulation of mitogenic signaling pathways in mesangial cells. AB - Although dietary fish oil supplementation has been used to prevent the progression of kidney disease in patients with IgA nephropathy, relatively few studies provide a mechanistic rationale for its use. Using an antithymocyte (ATS) model of mesangial proliferative glomerulonephritis, we recently demonstrated that fish oil inhibits mesangial cell (MC) activation and proliferation, reduces proteinuria, and decreases histologic evidence of glomerular damage. We therefore sought to define potential mechanisms underlying the antiproliferative effect of docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), the predominant omega 3 polyunsaturated fatty acids found in fish oil, in cultured MC. DHA and EPA were administered to MC as bovine serum albumin fatty-acid complexes. Low-dose (10-50 micromol/L) DHA, but not EPA, inhibited basal and epidermal growth factor (EGF) stimulated [(3)H]-thymidine incorporation in MCs. At higher doses (100 micromol/L), EPA and DHA were equally effective in suppressing basal and EGF stimulated MC mitogenesis. Low-dose DHA, but not EPA, decreased ERK activation by 30% (P <.01), as assessed with Western-blot analysis using phosphospecific antibodies. JNK activity was increased by low-dose DHA but not by EPA. p38 activity was not significantly altered by DHA or EPA. Cyclin E activity, as assessed with a histone H1 kinase assay, was inhibited by low-dose DHA but not by EPA. DHA increased expression of the cell cycle inhibitor p21 but not p27; EPA had no effect on p21 or p27. We propose that the differential effect of low-dose DHA vs EPA in suppressing MC mitogenesis is related to down-regulation of ERK and cyclin E activity and to induction of p21. PMID- 12761477 TI - Oncogenic ras induces gastrin/CCKB receptor gene expression in human colon cancer cell lines LoVo and Colo320HSR. AB - Gastrin has the ability to stimulate cell growth in some colorectal cancer cells and some of these cells also express gastrin/CCKB receptors, suggesting that gastrin and its autocrine loop are involved in their proliferation. We previously reported that oncogenic ras induced gastrin gene expression in colon cancer cells. The aim of this study was to investigate whether oncogenic ras also induces gastrin/CCKB receptor gene expression. A transiently transfected activated ras vector stimulated gastrin/CCKB receptor transcriptional activities in both Colo320HSR and LoVo cells, but these ras-increased activities were inhibited by a specific MEK inhibitor, PD98059. An RPA demonstrated that activated ras increased endogenous gastrin/CCKB receptor mRNA levels and PD98059 decreased them in LoVo cells. These findings suggest that oncogenic ras induces gastrin/CCKB receptor gene expression through some intracellular signaling pathways, including MEK, in colon cancer cell lines. PMID- 12761476 TI - Capacity of neutrophils and monocytes from human immunodeficiency virus-infected patients and healthy controls to inhibit growth of Mycobacterium bovis. AB - We compared the differences in growth inhibition of Mycobacterium bovis by monocytes and neutrophils from human immunodeficiency virus (HIV)-infected persons (n = 12; mean CD4 count = 451/mm(3)) and healthy controls (n = 6). Phagocytes from all HIV-infected patients were incubated with or without exogenous granulocyte-macrophate colony-stimulating factor (GMCSF; 500-1000 U/mL). In two of the HIV-infected patients, phagocytes were incubated with or without interleukin (IL)-2 or IL-8 (500-1000 U/mL). Compared with that in HIV infected patients, the reduction of M. bovis growth at 24 hours was 81% greater among monocytes and 69% greater among neutrophils from healthy controls (P =.03 and.04, respectively). Among HIV-infected patients, we noted greater mycobacterial reduction in monocytes (49%, P =.04) and neutrophils (42%, P =.05) from the early-stage patients (mean CD4 count = 760/mm(3)) compared with that in late-stage patients (mean CD4 count = 172/ mm(3)). Incubation with GM-CSF, IL-2, or IL-8 did not augment mycobactericidal activity. These findings suggest that the capacity of neutrophils and monocytes from HIV-infected patients to inhibit the growth of M. bovis is impaired, and this impairment is more pronounced in later stages of HIV infection. PMID- 12761478 TI - Isolation and identification of mesenchymal stem cells from human fetal pancreas. AB - Mesenchymal stem cells (MSCs) have been cultured from many sources, including bone marrow and liver. To further support our hypothesis that MSCs exist in most postnatal tissues, we isolated a clonogenic, multipotent, rapidly proliferating population of cells from a fetal pancreas and termed them "pancreas-derived mesenchymal stem cells" (PMSCs). They withstood being passaged as many as 30 times without sustaining significant structural changes. In this study, we showed that PMSCs are positive for CD44, CD29, and CDI3 but negative for CD34 and HLA-DR and that they stained with collagen I and III but not with von Willebrand factor antibody. During the log phase of growth, PMSCs proliferated, doubling in population in about 30 hours. Cell-cycle analysis showed that more than 90% of cells were in the G0 and G1 phases, whereas a small subpopulation of cells were actively engaged in proliferation (S + G2 + M = 3.55%). Under differentiation culture conditions, PMSCs differentiated into cells of osteogenic, chondrogenic, and adipogenic lineages. These results demonstrate that PMSCs can be isolated from human fetal pancreas by means of their adherent ability and that they are capable of self-renewal, propagation, and multipotent differentiation. PMID- 12761480 TI - Beautiful minds. PMID- 12761479 TI - Integrin alphaIIbbeta3 and shear-dependent action of glycoprotein Ibalpha stimulate platelet-dependent thrombin formation in stirred plasma. AB - Under conditions of arterial-wall shear rates, platelets bind to von Willebrand factor (vWf) by way of the glycoprotein Ib (GP Ib) complex and integrin alpha(IIb)beta(3). Both adhesive receptors may also play roles in the development of procoagulant activity of platelets. Here, we investigated the effect of shear stress, as provided by a rotating cylinder, on GP Ib- and integrin alpha(IIb)beta(3)-dependent thrombin generation in coagulating platelet-rich plasma (PRP). We measured thrombin continuously with the use of fluorometry from the cleavage rate of a fluorescent low-affinity substrate. The integrin alpha(IIb)beta(3) antagonist abciximab progressively reduced the peak of thrombin formation up to 43% when rate of stirring and shear stress were increased (estimated shear rates of 105-420 s(-1)). Abciximab did not lower the peak of thrombin formation in stirred PRP from patients with Glanzmann's thrombasthenia lacking alpha(IIb)beta(3) but, surprisingly, shortened the time until onset. In PRP from control subjects, antibodies specifically directed against vWf-binding epitopes on GP Ibalpha reduced thrombin formation, with 25% to 30% at the high but not at the low stirring rate. In combination with the anti-GP Ib antibody, abciximab retained its strong inhibitory effect only at the high stirring rate. We conclude that thrombin formation and coagulation in stirred PRP depend, to a large extent, on platelet adhesion to integrin alpha(IIb)beta(3) and, in a shear dependent way, on GP Ib. PMID- 12761482 TI - How effective is your patient teaching? PMID- 12761481 TI - WOCN celebrates leaders. PMID- 12761483 TI - Does fluid intake influence the risk for urinary incontinence, urinary tract infection, and bladder cancer? PMID- 12761484 TI - Venous leg ulcer care: how evidence-based is nursing practice? AB - OBJECTIVES: The objectives of this study were to (1) determine how congruent community-provided leg ulcer care was with best practice for venous leg ulcers and (2) identify organizational and clinical factors associated with the provision of best practice for venous leg ulcers. DESIGN: The practice variation study group was an audit of nursing agency client records to determine the provision of care. SETTING AND SUBJECTS: The study population was a home care cohort of persons with venous leg ulcers (n = 66) who received care from one Ontario home care nursing agency between March 1999 and November 1999. INSTRUMENT: The audit tool was developed with a checklist reflecting the common recommendations from 3 international practice guidelines, as well as organizational and clinical factors that may influence or reflect best practice. RESULTS: Half of client records (35/66) included an identified etiology of the leg ulcer. An Ankle Brachial Pressure Index score was documented prior to the initiation of compression on fewer than half of the records (21/44). Regular ulcer measurement was done for 11% of the clients (7/64). Two thirds of the clients (44/66) were treated with compression. More than 60% of the clients (40/66) had been seen by either a dermatologist or a vascular surgeon. Topical antibiotics were prescribed for two thirds of clients (44/64). Fifteen percent of clients (10/66) were assessed for pain, and 17% (11/66) received some form of pain management. Documentation of client education specific to the leg ulcer was present on 3% of records (2/66). The mean number of different nurses providing leg ulcer care to each client was 19. A registered practical nurse was the classification of nurse assigned to 43.8% (29/66) of the clients. CONCLUSION: Several gaps were identified in the care provided. A standardized approach to care is needed that includes a comprehensive leg ulcer assessment to determine the ulcer etiology, determination of an Ankle Brachial Pressure Index score to screen for the presence of arterial disease, and compression for all clients who meet the criteria for venous disease. A reorganization of services is required, which includes an increased role for community nurses in leg ulcer assessment and management. Organizational and clinical factors influencing the delivery of best practice need to be identified and addressed. PMID- 12761485 TI - Pressure ulcers and lateral rotation beds: a case study. AB - During a 6-month period, the WOC nurses at a 500-bed medical treatment facility noticed the development of nosocomial pressure ulcers on the sacrum, occiput, and heel areas of patients who were placed on lateral rotation specialty beds because they had pulmonary disorders. Measures were taken to address the problem by repositioning the patients and through a staff education program. Repositioning included repositioning the patient's head every 2 hours, thorough skin assessments every 2 hours, and ensuring that the patient's heels were subject to zero pressure. Staff education centered on the importance of using a risk assessment tool (the Braden scale) and understanding the clinical uses for lateral rotation beds. During the subsequent 6 months, the incidence of hospital acquired pressure ulcers decreased by 52%. Efforts to further decrease the number of pressure ulcers related to the use of lateral rotation beds continue. Issues such as length of stay on the bed and the appropriateness of manufacturer's guidelines still need to be addressed at this facility. This case study highlights the potential issues associated with lateral rotation beds and identifies the need for further research. PMID- 12761486 TI - Managing recurrent urinary catheter blockage: problems, promises, and practicalities. AB - Long-term urinary catheterization is rarely completely free of complications, and encrustation by mineral salts leading to catheter blockage is common in around 40% to 50% of long-term catheterized patients. Recurrent blockage is a problem, which is both distressing to patients and caregivers and costly to health services in terms of time and resources. This article addresses the causes of recurrent urinary catheter blockage, proactive approaches to care, and the evidence for use of catheter maintenance solutions to reduce buildup of mineral deposits. PMID- 12761487 TI - Case study: challenges of pessary management. AB - Genital prolapse is the relaxation of the supporting structures of the pelvic floor. Significant morbidity can be associated if left untreated. Patients can elect to have surgical repair of their prolapse or use a pessary. The more significant the pelvic organ prolapse the more difficult it is to manage with pessary support. The case study in this article describes such a patient and the challenges we faced with managing her advanced genital prolapse. PMID- 12761488 TI - Managing a highly exudative wound adjacent to an ileostomy. PMID- 12761489 TI - Response of Ewing tumor cells to forced and activated p53 expression. AB - The EWS-FLI1 transcription factor is consistently expressed in 85% of Ewing tumors (EFT). In heterologous cells, EWS-FLI1 induces p53-dependent cell cycle arrest or apoptosis. It has been speculated that the p53 tumor suppressor pathway may be generally compromised in EFT despite only rare p53 mutations. In order to test for functional integrity of this pathway, we have investigated a series of EFT cell lines that differ from each other with respect to their endogenous p53 and INK4A gene status for their response to ectopic p53 expression and to stimulation of endogenous p53 activity by X-ray treatment. Significant interindividual and intratumoral variations in the apoptotic propensity of EFT cell lines to transient expression of ectopic p53 were observed, which was independent of the level of p53 expression. In cell lines with a low apoptotic incidence, apoptosis was delayed and the surviving fraction showed a prolonged growth arrest. Complete resistance to p53-induced apoptosis in two cell lines established from the same patient was associated with a high BCL2/BAX ratio and low levels of APAF1. Sensitivity to X-rays showed a trend towards a higher apoptotic rate in wild-type (wt) p53 expressing than in p53 mutant cells. However, one wt p53-expressing EFT cell line was completely refractory to irradiation-stimulated cell death despite high apoptotic responsiveness to ectopic p53. No difference in Ser15 phosphorylation and the transcriptional activation of p53 targets was observed in wt p53 EFT cell lines irrespective of the induction of cell death or growth arrest. All together, our results demonstrate that despite significant variability in the outcome, cell death or cell cycle arrest, the p53 downstream pathway and the DNA damage signaling pathway are functionally intact in EFT. PMID- 12761490 TI - HER2/PI-3K/Akt activation leads to a multidrug resistance in human breast adenocarcinoma cells. AB - Growth factor receptor-mediated signal transduction has been implicated in conferring resistance to conventional chemotherapy on cancer cells. In this study, we delineated a pathway that involves HER2/PI-3K/Akt in mediating multidrug resistance in human breast cancer cells. We found that the cell lines that express both HER2 and HER3 appear to have a higher phosphorylation level of Akt (activated Akt). Transfection of HER2 in MCF7 breast cancer cells that express HER3 caused a phosphoinoside-3 kinase (PI-3K)-dependent activation of Akt, and was associated with an increased resistance of the cells to multiple chemotherapeutic agents (paclitaxel, doxorubicin, 5-fluorouracil, etoposide, and camptothecin). Selective inhibition of PI-3K or Akt activity with their respective dominant-negative expression vectors sensitized the cells to the induction of apoptosis by the chemotherapeutic agents. We further demonstrated that MCF7 cells expressing a constitutively active Akt, in which the phospholipid interactive PH domain of Akt was replaced by a farnesylation sequence for constitutive membrane anchorage (DeltaPH-Akt1-farn), showed a similar increased resistance to the chemotherapeutic agents. Our results suggest that activation of Akt1 by HER2/PI-3K plays an important role in conferring a broad-spectrum chemoresistance on breast cancer cells and that Akt may therefore be a novel molecular target for therapies that would improve the outcome of patients with breast cancer. PMID- 12761491 TI - The hypoxia-inducible factor-1 alpha is a negative factor for tumor therapy. AB - Tumor hypoxia negatively regulates cell growth and causes a more malignant phenotype by increasing the expression of genes encoding angiogenic, metabolic and metastatic factors. Of clinical importance, insufficient tumor oxygenation affects the efficiency of chemotherapy and radiotherapy by poorly understood mechanisms. The hypoxia-inducible factor (HIF)-1 is a master transcriptional activator of oxygen-regulated genes and HIF-1 is constitutively upregulated in several tumor types. HIF-1 might thus be implicated in tumor therapy resistance. We found that transformed mouse embryonic fibroblasts deficient for HIF-1alpha are more susceptible to the treatment with carboplatin, etoposide and ionizing radiation than wild-type cells. Increased cell death in HIF-1alpha-deficient cells was because of apoptosis and did not involve p53 induction. Tumor chemotherapy of experimental fibrosarcoma in immunocompromised mice with carboplatin and etoposide confirmed the enhanced susceptibility of HIF-1alpha deficient cells. Agents that did not cause DNA double-strand breaks, such as DNA synthesis inhibitors or a DNA single-strand break-causing agent equally impaired cell growth, independent of the HIF-1alpha genotype. Functional repair of a fragmented reporter gene was decreased in HIF-1alpha-deficient cells. Thus, hypoxia-independent basal HIF-1alpha expression in tumor cells, as known from untransformed embryonic stem cells, is sufficient to induce target gene expression, probably including DNA double-strand break repair enzymes. PMID- 12761493 TI - Function of p73, not of p53, is inhibited by the physical interaction with RACK1 and its inhibitory effect is counteracted by pRB. AB - The newly identified p53-related gene, p73, encodes a nuclear transcription factor. Unlike p53, p73 has various isoforms with different NH(2)- and COOH terminal tails. p73alpha with the longest COOH-terminal extension is most abundantly expressed in many tissues and cells among those splicing isoforms of p73 and the COOH-terminal region appears to have an autoregulatory function. To isolate and characterize the cellular protein(s) that interacts with the unique COOH-terminal region of p73alpha, we employed a yeast two-hybrid screen with a human fetal brain and 293 cell cDNA libraries. We identified the receptor for activated C kinase (RACK1) as a new member of p73alpha-binding proteins. The interaction was confirmed by coimmunoprecipitation experiments, whereas RACK1 did not interact with p53 or p73beta. Ectopic overexpression of RACK1 in SAOS-2 cells reduced the p73alpha-mediated transcription from the p53/p73-responsive promoters, and inhibited the p73alpha-dependent apoptosis. On the other hand, the p53-dependent transcriptional activation as well as apoptosis was unaffected in the presence of RACK1. Furthermore, we found that pRB physically bound to RACK1, and repressed the RACK1-dependent inhibition of p73alpha. Taken together, our observations suggest that pRB diminishes the RACK1-mediated inhibition of p73alpha activity through the interaction with RACK1. PMID- 12761492 TI - Differential regulation of SOCS genes in normal and transformed erythroid cells. AB - The SOCS family of genes are negative regulators of cytokine signalling with SOCS 1 displaying tumor suppressor activity. SOCS-1, CIS and SOCS-3 have been implicated in the regulation of red blood cell production. In this study, a detailed examination was conducted on the expression patterns of these three SOCS family members in normal erythroid progenitors and a panel of erythroleukemic cell lines. Unexpectedly, differences in SOCS gene expression were observed during maturation of normal red cell progenitors, viz changes to CIS were inversely related to the alterations of SOCS-1 and SOCS-3. Similarly, these SOCS genes were differentially expressed in transformed erythoid cells - erythroleukemic cells immortalized at an immature stage of differentiation expressed SOCS-1 and SOCS-3 mRNA constitutively, whereas in more mature cell lines SOCS-1 and CIS were induced only after exposure to erythropoietin (Epo). Significantly, when ectopic expression of the tyrosine kinase Lyn was used to promote differentiation of immature cell lines, constitutive expression of SOCS-1 and SOCS-3 was completely suppressed. Modulation of intracellular signalling via mutated Epo receptors in mature erythroleukemic lines also highlighted different responses by the three SOCS family members. Close scrutiny of SOCS-1 revealed that, despite large increases in mRNA levels, the activity of the promoter did not alter after erythropoietin stimulation; in addition, erythroid cells from SOCS-1-/- mice displayed increased sensitivity to Epo. These observations indicate complex, stage-specific regulation of SOCS genes during normal erythroid maturation and in erythroleukemic cells. PMID- 12761494 TI - Role of NF-kappaB and Akt/PI3K in the resistance of pancreatic carcinoma cell lines against gemcitabine-induced cell death. AB - Pancreatic cancer is resistant to almost all cytotoxic drugs. Currently, gemcitabine appears to be the only clinically active drug but, because of pre existing or acquired chemoresistance of most of the tumor cells, it failed to significantly improve the outcome of pancreatic carcinoma patients. The current study examined the relevance of nuclear factor kappaB (NF-kappaB) and PI3K/Akt in the resistance of five pancreatic carcinoma cell lines towards gemcitabine. Treatment for 24 h with gemcitabine (0.04-20 micro M) led to a strong induction of apoptosis in PT45-P1 and T3M4 cells but not in BxPc-3, Capan-1 and PancTu-1 cells. These resistant cell lines exhibited a high basal NF-kappaB activity in contrast to the sensitive cell lines. Furthermore, gemcitabine showed a dose dependent induction of NF-kappaB. At a dose of 0.04 micro M, gemcitabine still induced apoptosis in the sensitive cell lines, but did not induce NF-kappaB. In addition, NF-kappaB inhibition by MG132, sulfasalazine or the IkappaBalpha super repressor strongly diminished the resistance against gemcitabine (0.04-20 micro M). In contrast to this obvious correlation between basal NF-kappaB activity and gemcitabine resistance, PI3K/Akt seems not to be involved in gemcitabine resistance of these cell lines. Neither did the basal Akt activity correlate with the sensitivity towards gemcitabine treatment, nor did the inhibition of PI3K/Akt by LY294002 alter gemcitabine-induced apoptosis. These results indicate that constitutive NF-kappaB activity confers resistance against gemcitabine and that modulation of this activity by pharmacological or genetic approaches may have therapeutical potential when combined with gemcitabine in the treatment of pancreatic carcinoma. PMID- 12761495 TI - c-Myc degradation induced by DNA damage results in apoptosis of CHO cells. AB - Although tripchlorolide (TC), a compound purified from a Chinese herb Tripterygium Wilfordii Hook, has been demonstrated to be a potent antitumor agent, its mechanisms of action are unknown. The present study shows that TC induces apoptosis of Chinese Hamster Ovary (CHO) cells. Most strikingly, TC was particularly potent in inducing apoptosis of the UV41 mutant CHO cells, which are deficient in the ERCC4 gene encoding a nucleotide excision repair protein. TC caused a higher level of DNA damage in UV41 cells than those in the wild-type CHO cells or EM9 cells, which are deficient in single-strand break repair. These results provided a critical link between apoptotic hypersensitivity and DNA damage in defective nucleotide excision repair pathway of UV41 cells by TC treatment. Further analysis showed that degradation of the c-Myc protein in TC treated UV41 cells was much stronger than those in the wild-type CHOAA8 and the EM9. A proteasome inhibitor, MG132, reduced both the degradation of c-Myc and apoptosis in TC-treated UV41 cells. Expression of exogenous c-Myc also inhibited apoptosis of TC-treated UV41 cells. These results indicate that c-Myc degradation induced by DNA damage in the presence of TC contributes to induction of apoptosis of UV41 cells. PMID- 12761496 TI - Paclitaxel increases p21 synthesis and accumulation of its AKT-phosphorylated form in the cytoplasm of cancer cells. AB - CKI p21 is a regulator of cellular responses to microtubule damage induced by drugs such as paclitaxel (PTX). It mediates the G1 4N arrest postactivation of the spindle assembly checkpoint and protects cancer cells against PTX-induced cytotoxicity. We demonstrated here that low doses of PTX that are unable to activate the spindle assembly checkpoint, upregulate p21 by a p53-dependent pathway and induce its translocation to the cytoplasm. This cytoplasmic accumulation of p21 resulted from an AKT-dependent p21 phosphorylation leading to an association of p21 with 14-3-3. Furthermore, the cytoplasmic p21 accumulation observed in PTX-treated cells was inhibited by LY 294002, a specific PI-3 kinase inhibitor or by the expression of a dominant-negative AKT mutant. However, the kinase activity of AKT was unchanged in PTX-treated cells, suggesting that low doses of PTX could regulate p21 phosphorylation via inhibition of its dephosphorylation. As a functional consequence, we found that cytoplasmic accumulation of the phosphorylated form of p21 prevents the inhibitory effect of p21, enabling these cells to escape to the p53-dependent Gl/S and G2/M checkpoints. PMID- 12761497 TI - High levels of luteinizing hormone analog stimulate gonadal and adrenal tumorigenesis in mice transgenic for the mouse inhibin-alpha-subunit promoter/Simian virus 40 T-antigen fusion gene. AB - Transgenic (TG) mice expressing the Simian virus 40 T-antigen under the control of the murine inhibin-alpha promoter (Inhalpha/Tag) develop granulosa and Leydig cell tumors at the age of 5-6 months, with 100% penetrance. When these mice are gonadectomized, they develop adrenocortical tumors. Suppression of gonadotropin secretion inhibits the tumorigenesis in the gonads of intact animals and in the adrenals after gonadectomy. To study further the role of luteinizing hormone (LH) in gonadal and adrenal tumorigenesis, a double TG mouse model was generated by crossing the Inhalpha/Tag mice with mice producing constitutively elevated levels of LH (bLHbeta-CTP mice). Our results show that in double TG mice (bLHbeta CTP/Inhalpha/Tag), gonadal tumorigenesis starts earlier and progresses faster than in Inhalpha/Tag mice. Both ovarian and testicular tumors were histologically comparable with the tumors found in Inhalpha/Tag mice. In addition, adrenal tumorigenesis was found in intact double TG females, but not in Inhalpha/Tag females. Inhibin-alpha and LH receptor (LHR) were highly expressed in tumorigenic gonadal tissues, and the elevated LH levels were shown to be associated with ectopic LHR and high inhibin-alpha expression in the female adrenals. We conclude that in the Inhalpha/Tag tumor mouse model, elevated LH levels act as a tumor promoter, advancing gonadal and adrenal tumorigenesis. PMID- 12761499 TI - Allelic phasing of a mouse chromosome 11 deficiency influences p53 tumorigenicity. AB - Most tumour suppressor genes (TSGs) have been found through linkage studies in cancer predisposed families where the mutations have a high penetrance, for example, the breast cancer genes BRCA1 and BRCA2. Loss of heterozygosity (LOH) analyses of sporadic breast tumours indicate that there are many other putative TSGs yet to be identified. One such locus is proximal to BRCA1 on human chromosome 17q21. In an attempt to isolate this putative TSG, we have assessed a portion of the orthologous region on mouse chromosome 11 for its tumorigenic potential using segmental haploidy in combination with a p53 mutation. Two populations of animals were studied, with the deleted region being either on the same (cis) or on the homologous chromosome (trans) to a targeted mutant p53 allele. The deficiency elevated the tumour susceptibility of p53 heterozygous mice and modified the tumour spectrum, but only when the deficiency was in trans with the p53 mutation. Even though the genotype of these mice is identical, allelic phasing affects both the tumour spectrum and progression. PMID- 12761498 TI - Glucocorticoid receptor functions as a potent suppressor of mouse skin carcinogenesis. AB - Glucocorticoids are effective inhibitors of epidermal proliferation and skin tumorigenesis. Glucocorticoids affect cellular functions via glucocorticoid receptor (GR), a well-known transcription factor. Recently, we generated skin targeted transgenic mice overexpressing GR under control of the keratin5 promoter (K5-GR mice). To test the hypothesis that GR plays a role as a tumor suppressor in skin, we bred K5-GR transgenic mice with Tg.AC transgenic mice, which express v-Ha-ras oncogene in the skin, and compared the susceptibility of F1 offspring to TPA-induced skin carcinogenesis. GR overexpression in the epidermis dramatically inhibited skin tumor development. In K5-GR/ras+ double transgenic mice papillomas developed later and the average number of tumors per animal was 15% (in males) and 40% (in females) of the number seen in wild type (w.t./ras+) littermates. In addition, the papillomas in w.t./ras+ animals were eight to nine times larger. GR overexpression resulted in a decrease in keratinocyte proliferation combined with a modest increase in apoptosis and differentiation of keratinocytes in K5-GR/ras+ papillomas. Our data clearly indicate that interference of GR transgenic protein with nuclear factor kappa B (NF-kappaB) transcription factor had resulted in NF kappaB blockage in K5-GR/ras+ tumors. We discuss the role of NF-kappaB blockage in tumor-suppressor effect of GR. PMID- 12761500 TI - Overexpression of the thymosin beta-4 gene is associated with malignant progression of SW480 colon cancer cells. AB - Thymosin beta-4 (Tbeta-4), a small peptide originally isolated from calf thymus, modulates the formation of F-actin microfilaments by sequestering the monomeric G actin. Recent studies have shown that overexpression of the Tbeta-4 gene occurs not only in many human carcinomas but also in the highly metastatic melanomas and fibrosarcomas. However, little is known about the specific growth advantages acquired by different tumors from this genetic abnormality. To address the above questions, Tbeta-4-overexpressing human colon carcinoma (SW480) cells were established by stable transfection and their phenotypic changes were monitored. We found that both the morphology and the cortical actin cytoskeleton of SW480 cells were altered by Tbeta-4 overexpression. Moreover, both cellular level and that distributed over the intercellular junctions of the E-cadherin were decreased in the Tbeta-4 overexpressers, which were accompanied by a twofold increase in their saturation densities. Meanwhile, these cells also exhibited an increased ability to form colonies in soft agar. Interestingly, a dramatic increase of growth rate was detected in the Tbeta-4 overexpressers, which might be attributed to an accelerated proliferation induced by c-Myc that was activated by nuclear beta-catenin. Finally, a motility increase of these cells was demonstrated by two independent migration assays, which was accompanied by an enhanced focal contact. Taken together, our data suggest that the drastic growth property and motility changes of the SW480 cells overexpressing Tbeta-4 gene are due mainly to a deregulated cell-cell adhesion arisen from the downregulation of E-cadherin, plus uncontrolled cell proliferation owing to the upregulation of beta-catenin, both resulted from a breakdown of actin microfilaments caused by the overexpression of this G-actin sequestering peptide. PMID- 12761501 TI - Large-scale identification and characterization of human genes that activate NF kappaB and MAPK signaling pathways. AB - We have carried out a large-scale identification and characterization of human genes that activate the NF-kappaB and MARK signaling pathways. We constructed full-length cDNA libraries using the oligo-capping method and prepared an arrayed cDNA pool consisting of 150 000 cDNAs randomly isolated from the libraries. For analysis of the NF-kappaB signaling pathway, we introduced each of the cDNAs into human embryonic kidney 293 cells and examined whether it activated the transcription of a luciferase reporter gene driven by a promoter containing the consensus NF-kappaB binding sites. In total, we identified 299 cDNAs that activate the NF-kappaB pathway, and we classified them into 83 genes, including 30 characterized activator genes of the NF-kappaB pathway, 28 genes whose involvement in the NF-kappaB pathways have not been characterized and 25 novel genes. We then carried out a similar analysis for the identification of genes that activate the MARK pathway, utilizing the same cDNA resource. We assayed 145 000 cDNAs and identified 57 genes that activate the MARK pathway. Interestingly, 27 genes were overlapping between the NF-kappaB and the MAPK pathways, which may indicate that these genes play cross-talking roles between these two pathways. PMID- 12761502 TI - FEV acts as a transcriptional repressor through its DNA-binding ETS domain and alanine-rich domain. AB - Although most Ets transcription factors have been characterized as transcriptional activators, some of them display repressor activity. Here we characterize an Ets-family member, the very specifically expressed human Fifth Ewing Variant (FEV), as a transcriptional repressor. We show that among a broad range of human cell lines, only Dami megakaryocytic cells express FEV. This nuclear protein binds to Ets-binding sites, such as that of the human ICAM-1 promoter. We used this promoter to demonstrate that FEV can repress both basal transcription and, even more strongly, ectopically Ets-activated transcription. We identified two domains responsible for FEV-mediated repression: the ETS domain, responsible for passive repression, and the carboxy-terminal alanine-rich domain, involved in active repression. In the Ets-independent LEXA system also, FEV acts as a transcriptional repressor via its alanine-rich carboxy-terminal domain. The mechanism by which FEV actively represses transcription is currently unknown, since FEV-triggered repression is not reversed by the histone deacetylase inhibitor trichostatin A. We also showed that long-term overexpression of FEV proteins containing the alanine-rich domain prevents cell clones from growing, whereas clones expressing a truncated FEV protein lacking this domain develop like control cells. This confirms the importance of this domain in FEV-triggered repression. PMID- 12761503 TI - Flavonoid quercetin sensitizes a CD95-resistant cell line to apoptosis by activating protein kinase Calpha. AB - We previously demonstrated that quercetin, a naturally occurring flavonoid with strong antioxidant properties, was able to enhance programmed cell death in HPB acute lymphoblastic leukemia (ALL) cell line, derived from a human tymoma, when associated with the agonistic anti-CD95 monoclonal antibody. Here, we report that HPB-ALL cells are normally resistant to CD95-mediated apoptosis, and quercetin is able to sensitize this cell line through a mechanism independent of its antioxidant properties. In fact, other compounds structurally and functionally similar to quercetin, when associated with anti-CD95 antibody did not induce any CD95-mediated apoptosis, still maintaining their antioxidant capacity. We found that quercetin effects are mediated by the activation of PKCalpha. Treatment of HPB-ALL cells with quercetin slightly decreased PKCalpha activity, but when the flavonoid was associated with anti-CD95, the kinase activity increased by 12-fold with respect to the treatment with quercetin. In addition, overexpression of PKCalpha induced programmed cell death in the absence of any additional stimulus, while a kinase-defective mutant of PKCalpha was ineffective. Our data confirm the involvement of specific PKC isoforms in CD95 signaling and suggest, for the first time, that quercetin targets this pathway increasing apoptogenic response in a cell line resistant to CD95-mediated apoptosis. PMID- 12761504 TI - The early response gene IEX-1 attenuates NF-kappaB activation in 293 cells, a possible counter-regulatory process leading to enhanced cell death. AB - The early response gene IEX-1 is involved in the regulation of cellular growth and survival, and its expression is related to stress-, growth- and death inducing signals. Addressing the role of IEX-1 in the promotion of apoptosis, we investigated the effect of IEX-1 on nuclear factor-kappaB (NF-kappaB) activation. Stably transfected HEK-293 cells conditionally overexpressing IEX-1 exhibit decreased levels of NF-kappaB activity, either basal or TNFalpha induced, as shown by gel-shift and luciferase reporter gene assay. Furthermore, activated p65 accumulated in the nuclei of 293 cells to a lower degree, if IEX-1 expression was increased. This inhibited NF-kappaB activation was preceded by an altered turnover of IkappaBalpha and phospho-IkappaBalpha. In addition, IEX-1 expression also inhibited the activity of the 26S-proteasome, as shown by a fluorometric proteasome assay. Conversely, disruption of IEX-1 expression in 293 cells by stable transfection with specific anti-IEX-1 hammerhead ribozymes increased NF kappaB activity, and accelerated the degradation of IkappaBalpha. Along with these opposite effects of IEX-1 expression and IEX-1 disruption on NF-kappaB activation, the sensitivity of 293 cells towards various apoptotic stimuli also changed. In contrast to ribozyme-transduced 293 cells that were significantly less sensitive to apoptosis, this sensitivity was enhanced if IEX-1 expression was increased. Our data suggest that IEX-1 - itself an NF-kappaB target gene - inhibits the activation of this transcription factor, and hereby may counteract the antiapoptotic potential of NF-kappaB. PMID- 12761505 TI - Brazil's R&D agenda. PMID- 12761506 TI - Trade war: what is it good for? PMID- 12761507 TI - Trade war looms as US launches challenge over transgenic crops. PMID- 12761508 TI - Iraqi looters spark alert over radiation risks. PMID- 12761509 TI - Animal studies hint at staying power of SARS. PMID- 12761511 TI - Spiralling costs dog comet mission. PMID- 12761510 TI - Lack of trust hampers hunt for weapons. PMID- 12761512 TI - Brazilian brain experts plan research village. PMID- 12761513 TI - Copied citations give impact factors a boost. PMID- 12761514 TI - Researchers divided over ethics of a ban on cloning. PMID- 12761517 TI - Scientific literacy: Clear as mud. PMID- 12761518 TI - Brazilian science: Under new management. PMID- 12761519 TI - US federal funding ban puts babies at risk. PMID- 12761520 TI - Embryos aren't essential to stem-cell research. PMID- 12761521 TI - Politics hindering SARS work. PMID- 12761527 TI - Vision: The retina's fancy tricks. PMID- 12761528 TI - Astronomy: New direction for gamma-rays. PMID- 12761530 TI - Gene expression: Silent clones speak up. PMID- 12761531 TI - RNA interference: Cereal adultery. PMID- 12761532 TI - Condensed-matter physics: Thermopower to the people. PMID- 12761533 TI - Alzheimer's disease: Mental plaque removal. PMID- 12761534 TI - Synthetic chemistry: A perfect fit. PMID- 12761536 TI - Mammalian microevolution: Rapid change in mouse mitochondrial DNA. PMID- 12761537 TI - High-temperature superconductors: Universal nodal Fermi velocity. PMID- 12761538 TI - Marine ecology: Spring algal bloom and larval fish survival. PMID- 12761539 TI - Ecology: Hunting and fox numbers in the United Kingdom. PMID- 12761542 TI - Polarization of the prompt gamma-ray emission from the gamma-ray burst of 6 December 2002. AB - Observations of the afterglows of gamma-ray bursts (GRBs) have revealed that they lie at cosmological distances, and so correspond to the release of an enormous amount of energy. The nature of the central engine that powers these events and the prompt gamma-ray emission mechanism itself remain enigmatic because, once a relativistic fireball is created, the physics of the afterglow is insensitive to the nature of the progenitor. Here we report the discovery of linear polarization in the prompt gamma-ray emission from GRB021206, which indicates that it is synchrotron emission from relativistic electrons in a strong magnetic field. The polarization is at the theoretical maximum, which requires a uniform, large-scale magnetic field over the gamma-ray emission region. A large-scale magnetic field constrains possible progenitors to those either having or producing organized fields. We suggest that the large magnetic energy densities in the progenitor environment (comparable to the kinetic energy densities of the fireball), combined with the large-scale structure of the field, indicate that magnetic fields drive the GRB explosion. PMID- 12761543 TI - Experimental entanglement purification of arbitrary unknown states. AB - Distribution of entangled states between distant locations is essential for quantum communication over large distances. But owing to unavoidable decoherence in the quantum communication channel, the quality of entangled states generally decreases exponentially with the channel length. Entanglement purification--a way to extract a subset of states of high entanglement and high purity from a large set of less entangled states--is thus needed to overcome decoherence. Besides its important application in quantum communication, entanglement purification also plays a crucial role in error correction for quantum computation, because it can significantly increase the quality of logic operations between different qubits. Here we demonstrate entanglement purification for general mixed states of polarization-entangled photons using only linear optics. Typically, one photon pair of fidelity 92% could be obtained from two pairs, each of fidelity 75%. In our experiments, decoherence is overcome to the extent that the technique would achieve tolerable error rates for quantum repeaters in long-distance quantum communication. Our results also imply that the requirement of high-accuracy logic operations in fault-tolerant quantum computation can be considerably relaxed. PMID- 12761544 TI - Real-time detection of electron tunnelling in a quantum dot. AB - Nanostructures in which strong (Coulomb) interactions exist between electrons are predicted to exhibit temporal electronic correlations. Although there is ample experimental evidence that such correlations exist, electron dynamics in engineered nanostructures have been observed directly only on long timescales. The faster dynamics associated with electrical currents or charge fluctuations are usually inferred from direct (or quasi-direct) current measurements. Recently, interest in electron dynamics has risen, in part owing to the realization that additional information about electronic interactions can be found in the shot noise or higher statistical moments of a direct current. Furthermore, interest in quantum computation has stimulated investigation of quantum bit (qubit) readout techniques, which for many condensed-matter systems ultimately reduces to single-shot measurements of individual electronic charges. Here we report real-time observation of individual electron tunnelling events in a quantum dot using an integrated radio-frequency single-electron transistor. We use electron counting to measure directly the quantum dot's tunnelling rate and the occupational probabilities of its charge state. Our results provide evidence in favour of long (10 micros or more) inelastic scattering times in nearly isolated dots. PMID- 12761545 TI - Spin entropy as the likely source of enhanced thermopower in Na(x)Co2O4. AB - In an electric field, the flow of electrons in a solid produces an entropy current in addition to the familiar charge current. This is the Peltier effect, and it underlies all thermoelectric refrigerators. The increased interest in thermoelectric cooling applications has led to a search for more efficient Peltier materials and to renewed theoretical investigation into how electron electron interaction may enhance the thermopower of materials such as the transition-metal oxides. An important factor in this enhancement is the electronic spin entropy, which is predicted to dominate the entropy current. However, the crucial evidence for the spin-entropy term, namely its complete suppression in a longitudinal magnetic field, has not been reported until now. Here we report evidence for such suppression in the layered oxide Na(x)Co2O4, from thermopower and magnetization measurements in both longitudinal and transverse magnetic fields. The strong dependence of thermopower on magnetic field provides a rare, unambiguous example of how strong electron-electron interaction effects can qualitatively alter electronic behaviour in a solid. We discuss the implications of our finding--that spin-entropy dominates the enhancement of thermopower in transition-metal oxides--for the search for better Peltier materials. PMID- 12761546 TI - 146Sm-142Nd evidence from Isua metamorphosed sediments for early differentiation of the Earth's mantle. AB - Application of the 147Sm-143Nd chronometer (half-life of 106 Gyr) suggests that large-scale differentiation of the Earth's mantle may have occurred during the first few hundred million years of its history. However, the signature of mantle depletion found in early Archaean rocks is often obscured by uncertainties resulting from open-system behaviour of the rocks during later high-grade metamorphic events. Hence, although strong hints exist regarding the presence of differentiated silicate reservoirs before 4.0 Gyr ago, both the nature and age of early mantle differentiation processes remain largely speculative. Here we apply short-lived 146Sm-142Nd chronometry (half-life of 103 Myr) to early Archaean rocks using ultraprecise measurement of Nd isotope ratios. The analysed samples are well-preserved metamorphosed sedimentary rocks from the 3.7-3.8-Gyr Isua greenstone belt of West Greenland. Our coupled isotopic calculations, combined with an initial epsilon 143Nd value from ref. 6, constrain the mean age of mantle differentiation to 4,460 +/- 115 Myr. This early Sm/Nd fractionation probably reflects differentiation of the Earth's mantle during the final stage of terrestrial accretion. PMID- 12761547 TI - Spontaneous emergence of leaders and followers in foraging pairs. AB - Animals that forage socially often stand to gain from coordination of their behaviour. Yet it is not known how group members reach a consensus on the timing of foraging bouts. Here we demonstrate a simple process by which this may occur. We develop a state-dependent, dynamic game model of foraging by a pair of animals, in which each individual chooses between resting or foraging during a series of consecutive periods, so as to maximize its own individual chances of survival. We find that, if there is an advantage to foraging together, the equilibrium behaviour of both individuals becomes highly synchronized. As a result of this synchronization, differences in the energetic reserves of the two players spontaneously develop, leading them to adopt different behavioural roles. The individual with lower reserves emerges as the 'pace-maker' who determines when the pair should forage, providing a straightforward resolution to the problem of group coordination. Moreover, the strategy that gives rise to this behaviour can be implemented by a simple 'rule of thumb' that requires no detailed knowledge of the state of other individuals. PMID- 12761548 TI - GSK-3alpha regulates production of Alzheimer's disease amyloid-beta peptides. AB - Alzheimer's disease is associated with increased production and aggregation of amyloid-beta (Abeta) peptides. Abeta peptides are derived from the amyloid precursor protein (APP) by sequential proteolysis, catalysed by the aspartyl protease BACE, followed by presenilin-dependent gamma-secretase cleavage. Presenilin interacts with nicastrin, APH-1 and PEN-2 (ref. 6), all of which are required for gamma-secretase function. Presenilins also interact with alpha catenin, beta-catenin and glycogen synthase kinase-3beta (GSK-3beta), but a functional role for these proteins in gamma-secretase activity has not been established. Here we show that therapeutic concentrations of lithium, a GSK-3 inhibitor, block the production of Abeta peptides by interfering with APP cleavage at the gamma-secretase step, but do not inhibit Notch processing. Importantly, lithium also blocks the accumulation of Abeta peptides in the brains of mice that overproduce APP. The target of lithium in this setting is GSK 3alpha, which is required for maximal processing of APP. Since GSK-3 also phosphorylates tau protein, the principal component of neurofibrillary tangles, inhibition of GSK-3alpha offers a new approach to reduce the formation of both amyloid plaques and neurofibrillary tangles, two pathological hallmarks of Alzheimer's disease. PMID- 12761549 TI - Caenorhabditis elegans early embryogenesis and vulval morphogenesis require chondroitin biosynthesis. AB - Defects in glycosaminoglycan biosynthesis disrupt animal development and can cause human disease. So far much of the focus on glycosaminoglycans has been on heparan sulphate. Mutations in eight squashed vulva (sqv) genes in Caenorhabditis elegans cause defects in cytokinesis during embryogenesis and in vulval morphogenesis during postembryonic development. Seven of the eight sqv genes have been shown to control the biosynthesis of the glycosaminoglycans chondroitin and heparan sulphate. Here we present the molecular identification and characterization of the eighth gene, sqv-5. This gene encodes a bifunctional glycosyltransferase that is probably localized to the Golgi apparatus and is responsible for the biosynthesis of chondroitin but not heparan sulphate. Our findings show that chondroitin is crucial for both cytokinesis and morphogenesis during C. elegans development. PMID- 12761550 TI - Chondroitin proteoglycans are involved in cell division of Caenorhabditis elegans. AB - Glycosaminoglycans such as heparan sulphate and chondroitin sulphate are extracellular sugar chains involved in intercellular signalling. Disruptions of genes encoding enzymes that mediate glycosaminoglycan biosynthesis have severe consequences in Drosophila and mice. Mutations in the Drosophila gene sugarless, which encodes a UDP-glucose dehydrogenase, impairs developmental signalling through the Wnt family member Wingless, and signalling by the fibroblast growth factor and Hedgehog pathways. Heparan sulphate is involved in these pathways, but little is known about the involvement of chondroitin. Undersulphated and oversulphated chondroitin sulphate chains have been implicated in other biological processes, however, including adhesion of erythrocytes infected with malaria parasite to human placenta and regulation of neural development. To investigate chondroitin functions, we cloned a chondroitin synthase homologue of Caenorhabditis elegans and depleted expression of its product by RNA-mediated interference and deletion mutagenesis. Here we report that blocking chondroitin synthesis results in cytokinesis defects in early embryogenesis. Reversion of cytokinesis is often observed in chondroitin-depleted embryos, and cell division eventually stops, resulting in early embryonic death. Our findings show that chondroitin is required for embryonic cytokinesis and cell division. PMID- 12761557 TI - Cracking the combination. PMID- 12761551 TI - Deficiency of the adaptor SLP-65 in pre-B-cell acute lymphoblastic leukaemia. AB - Acute lymphoblastic leukaemia (ALL) is the commonest form of childhood malignancy, and most cases arise from B-cell clones arrested at the pre-B-cell stage of differentiation. The molecular events that arrest pre-B-cell differentiation in the leukaemic pre-B cells have not been well characterized. Here we show that the differentiation regulator SLP-65 (an adaptor protein also called BLNK or BASH) inhibits pre-B-cell leukaemia in mice. Reconstitution of SLP 65 expression in a SLP-65-/- pre-B-cell line led to enhanced differentiation in vitro and prevented the development of pre-B-cell leukaemia in immune-deficient mice. Tyrosine 96 of SLP-65 was required for this activity. The murine SLP-65-/- pre-B-cell leukaemia resembles human childhood pre-B ALL. Indeed, 16 of the 34 childhood pre-B ALL samples that were tested showed a complete loss or drastic reduction of SLP-65 expression. This loss is probably due to the incorporation of alternative exons into SLP-65 transcripts, leading to premature stop codons. Thus, the somatic loss of SLP-65 and the accompanying block in pre-B-cell differentiation might be one of the primary causes of childhood pre-B ALL. PMID- 12761559 TI - Genetic influence of CXCR6 chemokine receptor alleles on PCP-mediated AIDS progression among African Americans. AB - CXCR6 is a chemokine receptor and the primary coreceptor in SIV infection. A single nucleotide polymorphism 1469G-->A, results in a nonconservative change in codon 3 (CXCR6-E3K) of the N-terminus of the coreceptor. To investigate the relation between the chemokine receptor CXCR6 genotype and progression to Pneumocystis carinii pneumonia (PCP) and from PCP to death, we clinically assessed and genotyped 805 individuals from an African-American injection drug using cohort in Baltimore, MD, USA, for this CXCR6-E3K polymorphism. The allele frequency of CXCR6-3K was high (44%) in African Americans and rare in European Americans (f<1%). Although time to AIDS and PCP was similar for all CXCR6 genotypes, the median survival time from PCP to death for the CXCR6-3E/E and CXCR6-3E/K genotype was 1.5 years compared to 3.1 years for the CXCR6-K/K genotype. Individuals homozygous or heterozygous for the CXCR6-3E allele were 5.6 times more likely to die a PCP-mediated AIDS-related death than were individuals homozygous for CXCR6-3K. This study shows an association between CXCR6 genotype and progression from PCP to death among African-Americans with HIV. We suggest that CXCR6 may play a role in late-stage HIV-1 infection and may alter the progression to death after initial infection with PCP. PMID- 12761560 TI - Evolution of the beta defensin 2 gene in primates. AB - With the aim of further investigating the molecular evolution of beta defensin genes, after having analysed beta defensin 1 (DEFB1) in humans and several nonhuman primate species, we have studied the evolution of the beta defensin 2 gene (DEFB2), which codifies for a peptide with antimicrobial and chemoattractant activity, in humans and 16 primate species. We have found evidence of positive selection during the evolution of orthologous DEFB2 genes at two points on a phylogenetic tree relating these primates: during the divergence of the platyrrhines from the catarrhines and during the divergence of the Cercopithecidae from the Hylobatidae, Great Apes and humans. Furthermore, amino acid variations in Old World Monkeys seem to centre either on residues that are involved in oligomerisation in the human molecule, or that are conserved (40-80%) in beta-defensins in general. It is thus likely that these variations affect the biological function of the molecules and suggest that their synthesis and functional analysis might reveal interesting new information as to their role in innate immunity. PMID- 12761561 TI - Analysis of an IL-10 polymorphism in idiopathic pulmonary fibrosis. AB - Idiopathic pulmonary fibrosis (IPF) is a chronic progressive fibrotic disorder of the lung parenchyma. We have demonstrated changes in IL-10 protein production by alveolar macrophages (AMs) from patients with IPF, which we hypothesise could be because of an IL-10 gene polymorphism. We have screened the coding sequence and 3' untranslated region of IL-10 for polymorphisms using single-standard conformational polymorphism analysis. A novel polymorphism was identified resulting in a G to A substitution of +43 nucleotides from the start codon changing glycine to arginine at amino acid 15 of the signal peptide sequence. We have introduced the signal peptide mutation into the IL-10 gene and compared secretion of the mutant and wild-type forms after transient transfection of COS-7 cells. Our studies showed that the signal peptide mutation did not have a significant effect on secretion at 24 h post-transfection (P=0.4529 by Mann Whitney test). However, by 48 h there are significantly lower levels of mutant IL 10 (P=0.0515). There were no differences in the level of cell-associated IL-10 at either 24 or 48 h (P=0.9296 and 0.4268). We suggest that the mutation could affect the efficiency of protein translocation and signal peptide cleavage resulting in lower levels of IL-10 protein secretion. PMID- 12761562 TI - Linkage and association between Plasmodium falciparum blood infection levels and chromosome 5q31-q33. AB - We have previously mapped a locus controlling Plasmodium falciparum blood infection levels (PFBI) to chromosome 5q31-q33. We genotyped 19 microsatellite markers on chromosome 5q31-q33 in a new sample of 44 pedigrees comprising 84 nuclear families and 292 individuals living in a P. falciparum endemic area. Using a nonparametric multipoint variance-component approach (by GENEHUNTER), we evidenced a peak of linkage close to D5S636 (P=0.0069), with a heritability of 0.46. Using a variance-component method for linkage-disequilibrium mapping of quantitative traits (by QTDT) and the Bonferroni correction for multiple testing, we further detected allelic association in the presence of linkage between blood infection levels and D5S487 (P=6 x 10(-5); P(c)=0.0011), which is located on the distal part of the peak. These results confirm the importance of chromosome 5q31 q33 in the genetic control of PFBI levels. PMID- 12761563 TI - Mannose-binding lectin gene polymorphism predicts hospital admissions for COPD infections. AB - Infection frequently causes exacerbations of chronic obstructive pulmonary disease (COPD). Mannose-binding lectin (MBL) is a pattern-recognition receptor that assists in clearing microorganisms. Polymorphisms in the MBL2 gene reduce serum MBL levels and are associated with risk of infection. We studied whether the MBL2 codon 54 B allele affected serum MBL levels, admissions for infective exacerbation in COPD and disease susceptibility. Polymorphism frequency was determined by PCR-RFLP in 200 COPD patients and 104 smokers with normal lung function. Serum MBL was measured as mannan-binding activity in a subgroup of 82 stable COPD patients. Frequency of COPD admissions for infective exacerbation was ascertained for a 2-year period. The MBL2 codon 54 B allele reduced serum MBL in COPD patients. In keeping, patients carrying the low MBL-producing B allele had increased risk of admission for infective exacerbation (OR 4.9, P(corrected)=0.011). No association of MBL2 genotype with susceptibility to COPD was detected. In COPD, serum MBL is regulated by polymorphism at codon 54 in its encoding gene. Low MBL-producing genotypes were associated with more frequent admissions to hospital with respiratory infection, suggesting that the MBL2 gene is disease-modifying in COPD. MBL2 genotype should be explored prospectively as a prognostic marker for infection risk in COPD. PMID- 12761564 TI - Interferon-alpha receptor-1 (IFNAR1) variants are associated with protection against cerebral malaria in the Gambia. AB - The chromosome 21q22.11 cytokine receptor cluster contains four genes that encode subunits of the receptors for the cytokines interleukin-10 and interferon-alpha, beta and -gamma that may have a role in malaria pathogenesis. A total of 15 polymorphic markers located within these genes were initially genotyped in 190 controls and 190 severe malaria cases from The Gambia. Two interferon-alpha receptor-1 (IFNAR1) gene SNPs (17470 and L168 V) showed evidence for an association with severe malaria phenotypes and were typed in a larger series of samples comprising 538 severe malaria cases, 338 mild malaria cases and 562 controls. Both the 17470-G/G and L168V-G/G genotypes were associated with protection against severe malaria, in general, and cerebral malaria, in particular (P=0.004 and 0.003, respectively). IFNAR1 diplotypes were then constructed for these two markers using the PHASE software package. The (17470-G L168V-G/17470-G L168V-G) diplotype was found to be associated with a reduced risk of cerebral malaria and the (17470-C L168V-C/17470-G L168V-G) diplotype with an increased risk of cerebral malaria (overall 3 x 2 chi(2)=12.8, d.f.=2, P=0.002 and 3 x 2 chi(2)=15.2, d.f.=2, P=0.0005, respectively). These data suggest a role for the type I interferon pathway in resistance to cerebral malaria. PMID- 12761565 TI - Polymorphisms in lymphotoxin alpha and CD14 genes influence TNFalpha production induced by Gram-positive and Gram-negative bacteria. AB - Improved understanding of how host genetic variation affects resistance to microbial pathogens could lead to better treatment and/or prevention of infectious diseases. The lymphotoxin alpha (LTA)+250 and CD14-159 polymorphisms are associated with differences in susceptibility or outcome to several infections. We stimulated peripheral blood mononuclear cells (PBMC) from 22 healthy individuals with purified lipopolysaccharide (LPS), heat-killed Escherichia coli or Streptococcus pneumoniae. TNF alpha intracellular protein levels were measured by flow cytometry and mRNA was quantitated by RT-PCR. TNF alpha mRNA levels were higher in LTA+250GG subjects after 4 h incubation with LPS compared with LTA+250AA (T test, P=0.001). In contrast, after 8 h incubation with S. pneumoniae, there was slightly more TNF alpha mRNA in cells from LTA+250AA subjects. After 4 h incubation with LPS or E. coli, CD14-159TT subjects had higher TNF alpha mRNA levels than CD14-159CC (P=0.05, 0.033, respectively). Neither polymorphism affected the proportion of cells expressing intracellular TNF alpha protein. This suggests that the polymorphisms affected transcription and that other regulatory mechanisms affect production of TNF alpha protein. The effect of these two polymorphisms on TNF alpha mRNA production is stimulus dependent, with opposite effects observed for Gram-positive and Gram-negative stimuli. PMID- 12761566 TI - Novel association suggests multiple independent QTLs within chromosome 5q21-33 region control variation in total humans IgE levels. AB - Asthma is a common, heterogeneous, complex disease accompanied by raised total and specific immunoglobulin-E (IgE) antibody levels. Despite numerous previous reports of linkage and association of asthma, atopy and serum IgE levels to genes within the 5q21-33 region, definitive, replicable results are still not available. We used the classical twin design to (i) estimate the relative contributions of genes and environment to variation in total IgE levels, (ii) assess genetic linkage, and (iii) examine allelic association of 11 microsatellite markers spanning the 5q21-33 region to total IgE. Variation in total IgE level was shown to be highly heritable (65%). Although evidence for linkage of the 11 microsatellites to IgE was not observed, the omnibus test of association, not confounded by population substructure, showed positive association of D5S393 and D5S673 to IgE. Genes in the vicinity of D5S673 include hepatitis A virus receptor (HAVCR-1) and IL-12B. Recently, the mouse orthologue of HAVCR-1, the T-cell membrane family of proteins, have been shown to be in strong association with expression of airway hyperactivity in a mouse model of human asthma and atopy. IL-12B subserves many proinflammatory functions and also induces B cells proliferation. PMID- 12761567 TI - Identification, structural characterization, and tissue distribution of Tsg-5: a new TNF-stimulated gene. AB - Using DDRT-PCR, we compared the mRNA content of untreated and TNF-treated mouse embryonic fibroblasts (MEFs). Among differentially represented fragments, we identified and cloned a novel TNF-stimulated gene named Tsg-5. This gene, mapped to mouse chromosome 14, has three exons that can be alternatively spliced giving rise to two mRNA species, one spanning three exons and another that skips the second exon. Analysis of full-length Tsg-5 cDNA revealed a potential start codon within exon 2 encoding an ORF of 40 amino-acids. No homology with known mouse or human sequences, neither at the nucleotide nor at the amino-acid level could be found in public databases. In MEFs, Tsg-5 is induced by tumor necrosis factor alpha (TNF) and IL-1 beta, albeit with distinct kinetics. TNF-induced Tsg-5 expression is NF-kappa B-dependent as it was inhibited by MG132, lactacystin, Bay 11-7083, and Bay 11-7085. Analysis of Tsg-5 expression in vivo revealed that the gene and its encoded polypeptide are constitutively expressed in the thymus and ovary, whereas, in LPS-treated mice, Tsg-5 mRNA can be detected in the spleen, lung, and brain. Our data suggest that Tsg-5 encodes a new, rare transcript, with a very tight regulation of expression and differential splicing. PMID- 12761568 TI - Osteopontin polymorphisms and disease course in multiple sclerosis. AB - Osteopontin (OPN), also known as early T-cell activating gene (Eta-1), has been recently shown to be a critical factor in the progression of experimental autoimmune encephalomyelitis, and perhaps multiple sclerosis (MS). Here we investigated whether the 327T/C, 795C/T, 1128A/G or 1284A/C single-nucleotide polymorphisms in the OPN gene were correlated with susceptibility or any of the several clinical end points in a cohort of 821 MS patients. Overall, we observed no evidence of genetic association between the OPN polymorphisms and MS. Although not reaching statistical significance, a modest trend for association with disease course was detected in patients carrying at least one wild-type 1284A allele, suggesting an effect on disease course. Patients with this genotype were less likely to have a mild disease course and were at increased risk for a secondary-progressive clinical type. PMID- 12761569 TI - Epistatic effects occurring among susceptibility and protective MHC genes in IgA deficiency. AB - Immunoglobulin A deficiency (IgAD), the most prevalent primary immunodeficiency in Caucasian populations, shows strong evidence of polygenic inheritance with several associated genes being located in the major histocompatibility complex (MHC). Our aims were to determine which previously described MHC associations were primary and not secondary to a decrease or an increase in other MHC haplotype frequencies, to study the genetic interactions between all disease associated MHC haplotypes and, finally, to ascertain the relative importance of protection vs susceptibility. A relative predispositional effect (RPE) study showed that in addition to the primary positive association of IgAD with HLA DRB1*0102, DR3/TNFa2b3, and DR7 carrying haplotypes, DRB1*1501 was a marker of a primary protective factor in the Spanish population. Our data also indicate that the combined presence in an individual of two MHC susceptibility haplotypes notably increases the predisposition to the disease and that DRB1*1501 positive haplotypes eliminate the susceptibility conferred by any other MHC haplotype. PMID- 12761570 TI - A role of HSPs in apoptosis through "protein triage"? PMID- 12761571 TI - The apoptosis database. AB - The apoptosis database is a public resource for researchers and students interested in the molecular biology of apoptosis. The resource provides functional annotation, literature references, diagrams/images, and alternative nomenclatures on a set of proteins having 'apoptotic domains'. These are the distinctive domains that are often, if not exclusively, found in proteins involved in apoptosis. The initial choice of proteins to be included is defined by apoptosis experts and bioinformatics tools. Users can browse through the web accessible lists of domains, proteins containing these domains and their associated homologs. The database can also be searched by sequence homology using basic local alignment search tool, text word matches of the annotation, and identifiers for specific records. The resource is available at http://www.apoptosis-db.org and is updated on a regular basis. PMID- 12761572 TI - Caspase-like activity in programmed nuclear death during conjugation of Tetrahymena thermophila. AB - Apoptosis, or programmed cell death, is common in a variety of eucaryotes, from unicellular protozoa to vertebrates. The ciliated protozoan Tetrahymena thermophila has a unique apoptosis-like nuclear death during conjugation, called programmed nuclear death. This death program involves nuclear condensation (pyknosis) and oligonucleosomal DNA fragmentation in the parental macronucleus. Subsequently, the condensed nucleus is entirely resorbed in the autophagosome. Here we demonstrate that caspase-8- and -9-like activity was detected, but no caspase-3-like activity, by in vitro assay during the nuclear resorption process, suggesting that caspase-like activity is associated with both programmed cell death and apoptosis-like nuclear death in Tetrahymena. The use of indicator dye to detect the loss of mitochondrial membrane potential suggested the uptake of mitochondria and the degenerating macronucleus by the autophagosome. An involvement of mitochondria in the programmed nuclear death is discussed. PMID- 12761573 TI - A Drosophila model to study the functions of TWIST orthologs in apoptosis and proliferation. AB - The twist gene has been characterized for its role in myogenesis in several species. In addition, in mammalian cultured cells, it has been shown that twist is a potential oncogene antagonizing p53-dependent apoptosis. To study, in vivo, the role of twist in apoptosis and proliferation, we constructed transgenic Drosophila lines allowing ectopic expression of different twist orthologs. We report that: (i) Drosophila twist induces apoptosis and activates the reaper promoter, (ii) nematode twist induces arrest of proliferation without apoptosis, and (iii) human twist retains its potentialities observed in mammalian cultured cells and antagonizes Drosophila p53-dependent apoptosis. In addition, we show that human twist is able to induce cell proliferation in Drosophila. Data suggest that the pathway by which human twist antagonizes Drosophila p53 could be conserved. These transgenic lines thus constitute a powerful tool to identify targets and modifiers of human twist. PMID- 12761574 TI - Zinc inhibits Bax and Bak activation and cytochrome c release induced by chemical inducers of apoptosis but not by death-receptor-initiated pathways. AB - Zinc has been known for many years to inhibit apoptosis but the mechanism remains unclear. Originally thought to inhibit an apoptotic endonuclease, zinc has subsequently been shown to inhibit steps earlier in the pathway. Since many additional steps in apoptosis have now been defined, we have re-evaluated the steps inhibited by zinc. In response to activation of the chemical-mediated death pathway by anisomycin, 0.3 mM zinc inhibited Bax and Bak activation, cytochrome c release, and all of the subsequent steps in apoptosis. In the receptor-mediated death pathway initiated by Fas or tumor necrosis factor, 3 mM zinc was required to inhibit apoptosis as judged by inhibition of caspase 3 activity and DNA digestion, but it failed to inhibit cytochrome c release, activation of Bax and Bak, or upstream signaling events in this pathway. These results are consistent with zinc selectively inhibiting activation of BH3-only proteins required in the chemical pathway but inhibiting downstream caspase activation in the death receptor pathway. PMID- 12761575 TI - The regulatory domain of protein kinase Ctheta localises to the Golgi complex and induces apoptosis in neuroblastoma and Jurkat cells. AB - This study investigates apoptotic effects of protein kinase C (PKC) delta and theta in neuroblastoma cells. 12-O-tetradecanoylphorbol-13-acetate induces apoptosis in SK-N-BE(2) neuroblastoma cells overexpressing PKCdelta or PKCtheta, but not PKC epsilon. The PKC inhibitor GF109203X does not suppress this apoptotic effect, suggesting that it is independent of the catalytic activity of PKC. The isolated catalytic domains of PKCdelta and PKCtheta or the regulatory domain (RD) of PKCtheta also induce apoptosis in neuroblastoma cells. The apoptotic responses are suppressed by caspase inhibition and by Bcl-2 overexpression. The PKCtheta RD induced apoptosis also in Jurkat cells. Colocalisation analysis revealed that the PKCtheta RD primarily localises to the Golgi complex. The C1b domain is required for this localisation and removal of the C1b domain results in a PKCtheta construct that does not induce apoptosis. This suggests that the PKCtheta RD has apoptotic activity and that Golgi localisation may be important for this effect. PMID- 12761576 TI - Involvement of death receptor Fas in germ cell degeneration in gonads of Kit deficient Wv/Wv mutant mice. AB - Kit and its ligand stem cell factor (SCF) play a fundamental role in hematopoiesis, melanogenesis and gametogenesis. Homozygous W(v) mutant mice with a mutation in kit show abnormalities in these cell lineages. Fas is a member of the death receptor family inducing apoptosis. In this study, we generated double mutant mice (W(v)/W(v):Fas(-/-)) and analyzed histologically their reproductive organs. In testes and ovaries of the double-mutant mice, testicular germ cells and oocytes were detected, respectively, whereas the same-aged W(v)/W(v) mice contained neither cells. In addition, inhibition of Kit signals by administration of anti-Kit mAb, which induces degeneration of testicular germ cells in vivo in wild-type mice, did not cause degeneration in Fas-deficient mice. In testicular germ cells of W(v)/W(v) mutant mice, an increase of Fas expression was observed in spermatogonia. Further, in vitro treatment with SCF was shown to downregulate Fas on fibroblasts expressing exogenous Kit through activation of PI3-kinase/Akt. All the results clearly indicate that Fas-mediated apoptosis is involved in germ cell degeneration accompanied by defects in Kit-mediated signals, and Kit signaling negatively regulates Fas-mediated apoptosis in vivo. PMID- 12761577 TI - Apoptotic and necrotic blebs in epithelial cells display similar neck diameters but different kinase dependency. AB - Apoptotic and necrotic blebs elicited by H(2)O(2) were compared in terms of dynamics, structure and underlying biochemistry in HeLa cells and Clone 9 cells. Apoptotic blebs appeared in a few minutes and required micromolar peroxide concentrations. Necrotic blebs appeared much later, prior to cell permeabilization, and required millimolar peroxide concentrations. Strikingly, necrotic blebs grew at a constant rate, which was unaffected throughout successive cycles of budding and detachment. At 1 microm diameter, the necks of necrotic and apoptotic blebs were almost identical. ATP depletion was discarded as a major factor for both types of bleb. Inhibition of ROCK-I, MLCK and p38MAPK strongly decreased apoptotic blebbing but had no effect on necrotic blebbing. Taken together, these data suggest the existence of a novel structure of fixed dimensions at the neck of both types of plasma membrane blebs in epithelial cells. However, necrotic blebs can be distinguished from apoptotic blebs in their susceptibility to actomyosin kinase inhibition. PMID- 12761578 TI - Induced HMGA1a expression causes aberrant splicing of Presenilin-2 pre-mRNA in sporadic Alzheimer's disease. AB - The aberrant splicing isoform (PS2V), generated by exon 5 skipping of the Presenilin-2 (PS2) gene transcript, is a diagnostic feature of sporadic Alzheimer's disease (AD). We found PS2V is hypoxia-inducible in human neuroblastoma SK-N-SH cells. We purified a responsible trans-acting factor based on its binding to an exon 5 fragment. The factor was identified as the high mobility group A1a protein (HMGA1a; formerly HMG-I). HMGA1a bound to a specific sequence on exon 5, located upstream of the 5' splice site. HMGA1a expression was induced by hypoxia and the protein was accumulated in the nuclear speckles with the endogenous splicing factor SC35. Overexpression of HMGA1a generated PS2V, but PS2V was repressed by cotransfection with the U1 snRNP 70K protein that has a strong affinity to HMGA1a. HMGA1a could interfere with U1 snRNP binding to the 5' splice site and caused exon 5 skipping. HMGA1a levels were significantly increased in the brain tissue from sporadic AD patients. We propose a novel mechanism of sporadic AD that involves HMGA1a-induced aberrant splicing of PS2 pre-mRNA in the absence of any mutations. PMID- 12761579 TI - Mitochondrial membrane potential regulates matrix configuration and cytochrome c release during apoptosis. AB - During apoptosis, the mitochondrial membrane potential (MMP) decreases, but it is not known how this relates to the apoptotic process. It was recently suggested that cytochrome c is compartmentalized in closed cristal regions and therefore, matrix remodeling is required to attain complete cytochrome c release from the mitochondria. In this work we show that, at the onset of apoptosis, changes in MMP control matrix remodeling prior to cytochrome c release. Early after growth factor withdrawal the MMP declines and the matrix condenses. Both phenomena are reversed by adding oxidizable substrates. In mitochondria isolated from healthy cells, matrix condensation can be induced by either denying oxidizable substrates or by protonophores that dissipate the membrane potential. Matrix remodeling to the condensed state results in cristal unfolding and exposes cytochrome c to the intermembrane space facilitating its release from the mitochondria during apoptosis. In contrast, when a transmembrane potential is generated due to either electron transport or a pH gradient formed by acidifying the medium, mitochondria maintain an orthodox configuration in which most cytochrome c is sequestered in the cristae and is resistant to release by agents that disrupt the mitochondrial outer membrane. PMID- 12761580 TI - TNF combined with IFN-alpha accelerates NF-kappaB-mediated apoptosis through enhancement of Fas expression in colon cancer cells. AB - Immunostaining and EMSA revealed that NF-kappaB was activated strongly by TNF/IFN alpha compared to TNF alone in a human colon adenocarcinoma cell line, RPMI4788. Although inhibition of activated NF-kappaB, by using an NF-kappaB decoy, reduced cell viability after treatment with TNF only, NF-kappaB decoy resulted in recovery of cell viability after TNF/IFN-alpha treatment. Caspase-3 activity was increased in cells induced by TNF/IFN-alpha, while suppression of caspase-3 activity was observed in cells transfected with NF-kappaB decoy and then treated by TNF/IFN-alpha. On the other hand, Fas expression was strongly enhanced by TNF/IFN-alpha, and inhibition of TNF/IFN-alpha-induced NF-kappaB activation, by using NF-kappaB decoy, decreased Fas expression. Cell viability and caspase-3 activity decreased in cells treated with TNF/IFN-alpha and anti-FasL antibody. Taken together, our findings suggest that activated NF-kappaB induced by the crosstalk between TNF and IFN-alpha is a novel pro-apoptotic signal acting via enhancement of Fas expression. PMID- 12761581 TI - A caspase-8-independent component in TRAIL/Apo-2L-induced cell death in human rhabdomyosarcoma cells. AB - Tumor necrosis factor related apoptosis inducing ligand (TRAIL) belongs to the Tumor necrosis factor (TNF) family of death-inducing ligands, and signaling downstream of TRAIL ligation to its receptor(s) remains to be fully elucidated. Components of the death-inducing signaling complex (DISC) and TRAIL signaling downstream of receptor activation were examined in TRAIL - sensitive and resistant models of human rhabdomyosarcoma (RMS). TRAIL ligation induced DISC formation in TRAIL-sensitive (RD, Rh18, Rh30) and TRAIL-resistant RMS (Rh28, Rh36, Rh41), with recruitment of FADD and procaspase-8. In RD cells, overexpression of dominant-negative FADD (DNFADD) completely abolished TRAIL induced cell death in contrast to dominant-negative caspase- 8 (DNC8), which only partially inhibited TRAIL-induced apoptosis, growth inhibition, or loss in clonogenic survival. DNC8 did not inhibit the cleavage of Bid or the activation of Bax. Overexpression of Bcl-2 or Bcl-xL inhibited TRAIL-induced apoptosis, growth inhibition, and loss in clonogenic survival. Bcl-2 and Bcl-xL, but not DNC8, inhibited TRAIL-induced Bax activation. Bcl-xL did not inhibit the early activation of caspase-8 (<4 h) but inhibited cleavage of Bid, suggesting that Bid is cleaved downstream of the mitochondria, independent of caspase-8. Exogenous addition of sphingosine also induced activation of Bax via a caspase-8-and Bid independent mechanism. Further, inhibition of sphingosine kinase completely protected cells from TRAIL-induced apoptosis. Data demonstrate that in RMS cells, the TRAIL signaling pathway circumvents caspase-8 activation of Bid upstream of the mitochondria and that TRAIL acts at the level of the mitochondria via a mechanism that may involve components of the sphingomyelin cycle. PMID- 12761582 TI - Lymphoma development in Bax transgenic mice is inhibited by Bcl-2 and associated with chromosomal instability. AB - Bax is a Bcl-2 family member that promotes apoptosis but has paradoxical effects on lymphoma development in p53-deficient mice. To better understand the mechanism of Bax-induced lymphoma development, the effect of Bax levels, p53 status and Bcl 2 coexpression on lymphoma development were determined. In addition, DNA content and cytogenetics were performed on young (premalignant) Lck-Bax mice as measures of genetic instability. Bax promoted lymphoma development in p53-deficient mice in a dose-dependent manner. Bax expression also led to lymphoma development in both p53 +/- and +/+ animals. Ploidy analysis in mice prior to the onset of overt thymic lymphomas demonstrated that Lck-Bax transgenic mice were more likely to be aneuploid and demonstrate increased chromosome instability. With tumor progression, aneuploidy increased and Bax expression was maintained. Importantly, coexpression of Bcl-2 delayed lymphoma development in Lck-Bax transgenic mice. These data support a model in which increased sensitivity to apoptosis leads directly to chromosome instability in developing T cells and may explain a number of paradoxical observations regarding Bcl-2 family members and the regulation of cancer. PMID- 12761583 TI - Identification of candidate genes encoding the core components of the cell death machinery in the Ciona intestinalis genome. PMID- 12761584 TI - Apoptosis and efficient repair of DNA damage protect human keratinocytes against UVB. PMID- 12761585 TI - Initial impact of a national dental education program on the oral health and dental knowledge of children. AB - Oral health educational programs have been reported to have a variable impact on the oral health status of program participants. This paper reports the impact of an educational oral health program conducted within a single Boys & Girls Club of America. The objective of this 4-week examiner-blind study was to determine the impact of the educational program on the gingival health (gingivitis and plaque) of participating children who were between the ages of 5 and 15. The multi-week program taught the participants the basics of oral biology and disease, as well as proper oral health prevention including oral hygiene, dietary modification, and the importance of visiting the dentist. A calibrated examiner measured whole mouth Loe-Silness Gingival Index (GI) and Turesky Modification of Quigley-Hein Plaque index (PI) at baseline (immediately prior to the initiation of the educational program) and 4 weeks later. The primary efficacy analysis was based on change from baseline for 75 subjects who were enrolled at baseline, participated in the educational program, and were examined 4 weeks later. Mean baseline GI score was 0.37, while the 4 week mean GI score was reduced to 0.18. This represents a 51% reduction in GI score with p<0.001. Mean baseline PI score was 3.80, while the 4 week mean PI score was reduced to 2.68. This represents a 29% reduction in PI score with p<0.001. In addition, subjects completed a questionnaire (5 questions) at baseline and at 4 weeks to assess their oral health knowledge. The subject population was found to have statistically significantly (p<0.05) greater knowledge with respect to optimal brushing time and optimal frequency of dental recall visits following the program at week 4. Collectively, these data support the role of the educational program in promoting improved oral health in these children over a one month period. PMID- 12761587 TI - Common errors on panoramic radiographs taken in a dental school. AB - The aim of this study is to evaluate the frequency of common errors seen on panoramic radiographs taken in the Radiology Department of a dental school by trained assistants. Four hundred and sixty radiographs were evaluated for 20 categories of common errors. Out of the evaluated radiographs, 37.61% were found to be error-free. The most common errors were found to be the palatoglossal airspace shadow of air above the tongue due to the patient not raising the tongue against the palate (46.30%) and the superimposition of hyoid bone with the mandible (26.30%) respectively. The least common error was found to be dirty or bent films (0.21%). The quality of panoramic radiographs could be enhanced by improving radiographic technique. PMID- 12761586 TI - Effects of a betaine-containing toothpaste on subjective symptoms of dry mouth: a randomized clinical trial. AB - Our aim was to study the effects of mildly flavoured sodium lauryl sulphate (SLS) containing and detergent-free toothpastes with and without betaine (BET) on subjective symptoms of dry mouth in a randomised clinical trial. BET is an osmoprotectant that reacts with molecules to supply the surface with a water coating that protects cells from surfactants. Twenty-seven xerostomic patients and 18 healthy controls took part in the randomised, double-blind clinical trial with a crossover design. Three mildly flavoured toothpastes: (1) 4% BET, (2) 1% SLS and 4% BET, and (3) 1% SLS were used for six weeks each. The reference or washout paste contained neither SLS nor BET. The subjects' dental appointments were at the beginning of the trial and before and after the use of each toothpaste. At each appointment, the subjects were interviewed about subjective sensations of dry mouth (Visual Assessment Scoring (VAS) Index). The subjects did not report any adverse effects in connection with the use of the toothpastes. The VAS scores for lip dryness and eating difficulties were significantly lower for the BET paste (lip dryness: BET30 cigarettes/day). Fifty-two percent of the alcoholics stated they frequently forgot to brush their teeth and 43% observed bleeding of their gums. The mean DMF-T was 20 (2 D-T, 9 M-T, 9 F-T), the mean QHI was 2.4, and the mean PBI was 1.8. Forty-nine percent of the sextants were scored CPI 3 or 4; about 25% were edentulous (CPI X). Our results indicate severe alcoholics have a high risk of periodontal break down and tooth loss. To what extent these findings were caused by general/oral neglect alone (in combination with nicotine abuse) is at present unknown. PMID- 12761590 TI - Cephalometric soft tissue profile analysis between two different ethnic groups: a comparative study. AB - The aim of this investigation was to study and compare the cephalometric soft tissue profile analysis between Saudis and Caucasian Americans. The study was carried out using standardized cephalometric radiographs of 56 Saudi subjects (30 males and 26 females) with pleasant and balanced facial profiles, competent lips, normal overjet and overbite, and showing no craniofacial deformities. Subject ages ranged from 22 to 23 years. One skeletal and thirteen soft tissue variables were investigated. F-test, two samples t-test, Mann-Whitney, and Wilcoxon tests were used for data analysis. The results showed no statistical significant differences between the Saudi males and females except for the angle of total facial convexity, soft tissue facial plane angle, lower lip length, sagittal nasal tip to the most protrusive lip distance, and also sagittal chin to the most protrusive lip distance. The Saudi females had a greater angle of total facial convexity and soft tissue facial plane angle than the males. In addition, the females had a shorter lower lip. They also had a short distance between the nasal tip and chin to the most protrusive lip. These results reveal significant differences in most of the soft tissue variables when comparing Saudis with Caucasian Americans as well as in other ethnic groups. Most of these variables are essential for the diagnosis and treatment planning of cases requiring orthodontics and orthognathic surgery. PMID- 12761591 TI - Benign fibrous histiocytoma: report of case. AB - Benign fibrous histiocytoma is a rare and usually painless oral neoplasm found in adults that may affect either soft tissue or bone typically noted in their fifth decade. This case was found in a 32 year-old Caucasian male who presented with a fairly well circumscribed unilocular radiolucent lesion extending from the mandibular right first incisor to the left first premolar and reaching the inferior mandible on a panoramic radiograph. A bony window was created and the intrabony lesion was curetted. Multiple sections revealed a cellular tumor composed of uniform spindle-shaped cells arranged in a prominent whorled or storiform pattern. Scattered xanthoma cells, multinucleated giant cells, lymphocytes, and deposits of hemosiderin were noted throughout the lesional stroma. Although malignant fibrous histiocytoma of the bone is relatively well known, benign fibrous histiocytoma of the bone is very rare. PMID- 12761592 TI - Assessment for personal and professional success. AB - This article provides two assessment strategies to help dental professionals determine how close they are to achieving total success in their personal life and dental practice. Total success is defined as achieving and balancing success in three categories: physical health, financial health, and emotional health. By rating answers to questions on a scale from 1 to 5, the reader can determine the level of success they have attained in the various areas of their personal life and in their dental practice. This information may be useful in identifying those areas that need improvement to achieve total and balanced success. Interpretation of the scores and strategies to develop those areas requiring improvement are discussed. PMID- 12761593 TI - The AAOMP case challenge: painful mandibular mass associated with a molar. PMID- 12761594 TI - Continuing education: child abuse and neglect: implications for the dental profession. PMID- 12761595 TI - Usefulness of lymphoscintigraphy and intraoperative gamma probe detection in the identification of sentinel nodes in cervical cancer. AB - The aim of our study was to prospectively assess the clinical usefulness of sentinel lymphoscintigraphy and intraoperative gamma probe detection in identifying sentinel nodes (SNs) in patients with early cervical carcinoma. Between 6 and 24 h before radical hysterectomy, lymphoscintigrams were obtained following peritumoural injection of technetium-99m antimony sulphur colloid in 26 patients (mean age 45 years, range 32-71 years) with cervical cancer (FIGO I IIa). Scanning for radioactive nodes was performed using a hand-held collimated gamma-detecting probe in the initial stages of the main operation, during and before exposure of the retroperitoneal spaces. After separation and removal of radioactive, blue nodes, specimens were submitted for pathological evaluation. SNs were successfully localised using a combination of lymphoscintigraphy and intraoperative gamma probe detection in all 26 cases. However, in two of the 26 cases, SNs were only localisable using the gamma-detecting probe. A histologically positive SN was found in only one case. It is concluded that, in cervical cancer, lymphoscintigraphy and SN biopsy using a gamma-detecting probe are easy and reliable methods for the detection of SNs and are of value in defining the necessity and extent of node dissection. PMID- 12761596 TI - A model-based method for the prediction of whole-body absorbed dose and bone marrow toxicity for 186Re-HEDP treatment of skeletal metastases from prostate cancer. AB - In high-activity rhenium-186 hydroxyethylidene diphosphonate ((186)Re-HEDP) treatment of bone metastatic disease from prostate cancer the dose-limiting factor is haematological toxicity. In this study, we examined the correlation of the injected activity and the whole-body absorbed dose with treatment toxicity and response. Since the best response is likely to be related to the maximum possible injected activity limited by the whole-body absorbed dose, the relationship between pre-therapy biochemical and physiological parameters and the whole-body absorbed dose was studied to derive an algorithm to predict the whole body absorbed dose prior to injection of the radionuclide. The whole-body retention of radioactivity was measured at several time points after injection in a cohort of patients receiving activities ranging between 2,468 MBq and 5,497 MBq. The whole-body absorbed dose was calculated by fitting a sequential series of exponential phases to the whole-body time-activity data and by integrating this fit over time to obtain the whole-body cumulated activity. This was then converted to absorbed dose using the Medical Internal Radiation Dose (MIRD) committee methodology. Treatment toxicity was estimated by the relative decrease in white cell (WC) and platelet (Plt) counts after the injection of the radionuclide, and by their absolute nadir values. The criterion for a treatment response was a 50% or greater decrease in prostate-specific antigen (PSA) value lasting for 4 weeks. Alkaline phosphatase (AlkPh), chromium-51 ethylene diamine tetra-acetate ((51)Cr-EDTA) clearance rate and weight were measured before injection of the radionuclide. The whole-body absorbed dose showed a significant correlation with WC and Plt toxicity ( P=0.005 and 0.003 for the relative decrease and P=0.006 and 0.003 for the nadir values of WC and Plt counts respectively) in a multivariate analysis which included injected activity, whole body absorbed dose, pre-treatment WC and Plt baseline counts, PSA and AlkPh values, and the pre-treatment Soloway score. The injected activity did not show any correlation with WC or Plt toxicity, but it did correlate with PSA response ( P=0.005). These results suggest that the administration of higher activities would be likely to generate a better response, but that the quantity of activity that can be administered is limited by the whole-body absorbed dose. We have derived and evaluated a model that estimates the whole-body absorbed dose on an individual patient basis prior to injection. This model uses the level of injected activity and pre-injection measurements of AlkPh, weight and (51)Cr-EDTA clearance. It gave good estimates of the whole-body absorbed dose, with an average difference between predicted and measured values of 15%. Furthermore, the whole-body absorbed dose predicted using this algorithm correlated with treatment toxicity. It could therefore be used to administer levels of activity on a patient-specific basis, which would help in the optimisation of targeted radionuclide therapy. We believe that algorithms of this kind, which use pre injection biochemical and physiological measurements, could assist in the design of escalation trials based on a toxicity-limiting whole-body absorbed dose, rather than using the more conventional activity escalation approach. PMID- 12761597 TI - Rapid normalization of osseous FDG uptake following traumatic or surgical fractures. AB - It is known that following a traumatic fracture or surgical intervention, bone scintigraphy reveals positive results for an extended period of time, posing a challenge when evaluating patients for possible malignancy or superimposed osteomyelitis. Previous reports indicate that acute fractures can also result in increased fluorine-18 fluorodeoxyglucose (FDG) accumulation and therefore cause difficulties when patients are evaluated for other indications by FDG-PET. The purpose of this study was to assess the pattern and time course of abnormal FDG uptake following traumatic or surgical fracture. A total of 1,517 consecutive patients who underwent whole-body FDG-PET imaging were retrospectively studied. A history of fractures or orthopedic intervention was obtained from an interview prior to scanning. The FDG-PET results were compared with the results of other imaging studies, including bone scans, radiographs, CT, and MRI, as well as surgical pathology reports. Thirty-seven patients with a known date of traumatic or surgical fracture were identified. Among these, 14 had fractures or surgery within 3 months prior to FDG-PET, while 23 had fractures or surgical intervention greater than 3 months prior to FDG-PET. FDG-PET showed no abnormally increased uptake at the known fracture or surgical sites in 30 of these patients. Notably, in the 23 patients with fractures more than 3 months old, all but one showed no abnormally increased uptake. Furthermore, the positive FDG uptake in this exception was a result of complicating osteomyelitis. In the 14 patients with a history of fracture less than 3 months old, only six had abnormally increased FDG uptake. Following traumatic or surgical fractures, FDG uptake is expected to be normal within 3 months unless the process is complicated by infection or malignancy. PMID- 12761598 TI - Imaging of renal carcinoma xenografts with 64Cu-labelled anti-L1-CAM antibody chCE7. PMID- 12761599 TI - Imaging of chronic recurrent multifocal osteomyelitis of childhood first presenting with isolated primary spinal involvement. AB - OBJECTIVE: Initial presentation with primary spinal involvement in chronic recurrent multifocal osteomyelitis of childhood (CRMO) is rare. Our objective was to review the imaging appearances of three patients who had CRMO who initially presented with isolated primary spinal involvement. DESIGN AND PATIENTS: The imaging, clinical, laboratory and histology findings of the three patients were retrospectively reviewed. Imaging included seven spinal MR imaging scans, one computed tomography scan, nine bone scans, two tomograms and 16 radiographs. These were reviewed by two musculoskeletal radiologists and a consensus view is reported. All three patients presented with atraumatic spinal pain and had extensive bone spinal pathology. The patients were aged 11, 13 and 12 years. There were two females and one male. RESULTS AND CONCLUSIONS: The initial patient had thoracic T6 and T8 vertebra plana. Bone scan showed additional vertebral body involvement. Follow-up was available over a 3 year period. The second patient had partial collapse of T9 and, 2 years later, of C6. Subsequently extensive multifocal disease ensued and follow-up was available over 8 years. The third patient initially had L3 inferior partial collapse and 1 year later T8 involvement with multifocal disease. Follow-up was available over 3 years. The imaging findings of the three patients include partial and complete vertebra plana with a subchondral line adjacent to endplates associated with bone marrow MR signal alterations. Awareness of the imaging appearances may help the radiologist to include this entity in the differential diagnosis in children who present with spinal pathology and no history of trauma. Histopathological examination excludes tumor and infection but with typical imaging findings may not always be necessary. PMID- 12761600 TI - Imaging in the diagnosis of symptomatic forearm muscle herniation. AB - Muscle herniation can be defined as protrusion of a portion of muscle through an acquired or congenital defect of enclosing fascia. Although it is usually a cosmetic problem, it can lead to local pain and tenderness after prolonged exertion. In this report, we present a case of flexor digitorum superficialis muscle herniation in a 58-year-old man. The radiographic, ultrasonographic and magnetic resonance imaging findings are described with dynamic examination, permitting demonstration of muscle herniation through the fascial defect during muscle contraction. PMID- 12761601 TI - MRI in methotrexate-related leukoencephalopathy: Disseminated necrotising leukoencephalopathy in comparison with mild leukoencephalopathy. AB - We report two fatal cases of methotrexate (MTX)-induced disseminated necrotising leukoencephalopathy (DNL) in which MRI was repeated from the onset. Initial T2 weighted images showed multiple areas of high signal, mainly in deep cerebral white matter, which on follow-up, spread and coalesced to involve the entire white matter. Small irregular low-signal foci on T2-weighted images were seen within the high-signal lesions. Multiple areas of contrast enhancement corresponded to these low-signal foci. The condition of both patients deteriorated and they died. We compared their MRI findings with those of seven patients with mild MTX-related leukoencephalopathy, six of whom were asymptomatic; one had transient neurological symptoms. They showed no contrast enhancement, but rather mild-to-moderate diffuse high signal in deep white matter, which later disappeared. These findings suggest that multiple low-signal foci on T2-weighted images with contrast enhancement may be characteristic of DNL, and that contrast-enhanced imaging is useful to differentiate this condition from mild leukoencephalopathy. PMID- 12761603 TI - Spontaneous bilateral subdural haematomas in the posterior cranial fossa revealed by MRI. AB - A 52-year-old woman treated for acute myeloproliferative disease developed progressive stupor. CT showed obstructive hydrocephalus resulting from unexplained mass effect on the fourth ventricle. MRI revealed bilateral extra axial collections in the posterior cranial fossa, giving high signal on T1- and T2-weighted images, suggesting subacute subdural haematomas. Subdural haematomas can be suspected on CT when there is unexplained mass effect. MRI may be essential to confirm the diagnosis and plan appropriate treatment. PMID- 12761602 TI - Endovascular treatment of experimentally induced aneurysms in rabbits using stents: a feasibility study. AB - Although Guglielmi detachable coil (GDC) systems have been generally accepted for treatment of intracranial aneurysms, primary stenting of aneurysms using porous stents or implantation of coils after stent placement remains experimental. Testing of these new methods requires an animal model which imitates human aneurysms in size, configuration and neck morphology. We assessed in detail the technical requirements of and steps for transfemoral stent treatment of experimentally induced aneurysms at the top of the brachiocephalic trunk in rabbits. We created aneurysms in ten rabbits by distal ligation and intraluminal digestion of the right common carotid artery with elastase. We treated five animals with porous stents alone, and five with stents plus coiling via the meshes of the stent, which permitted dense packing of coils. No complications related to the procedures occurred. In all animals, even in those treated solely with porous stents, total occlusion of the aneurysm was achieved. Our animal model can be suitable for testing the biocompatibility and occlusion rate of new methods and devices for the treatment of experimental aneurysms. PMID- 12761604 TI - Pilot study on the feasibility and acceptability of academic detailing in general practice. AB - OBJECTIVE: To investigate the feasibility and acceptability of academic detailing in general practice in the North-East-Flanders region of Belgium. METHODS: All local quality groups in the study region were randomly allocated to either (1) two academic detailing visits to the individual members or (2) two academic detailing visits during the meetings of the local quality group. During these visits, the results of a systematic review of the literature on non-steroidal anti-inflammatory agents (NSAIDs) were presented. RESULTS: Of 14 local quality groups, 12 allocated to the individual intervention agreed to participate. Of the 184 physicians invited to receive an academic detailer, 142 had two visits on NSAIDs. One hundred and five physicians gave their opinion on the visits by means of a questionnaire. The great majority of the responders (90%) wished to receive an academic detailer on other topics in the future, with a frequency of two visits per year. Of 13 local quality groups, 12 allocated to the group intervention, with a total number of 192 physicians who agreed to participate. Ten local quality groups with 166 members received the information on NSAIDs. The actual attendance rate per intervention session was approximately two out of three physicians. All 166 physicians of the participating local quality groups were offered an evaluation form, and 79 responded. Of the responders who received the full group intervention, 88% wished to have more academic detailing visits on other topics in their local quality group. A comparison between the evaluation forms of the individual and the group intervention revealed no major differences concerning the acceptability of the intervention. CONCLUSION: The pilot study showed that academic detailing is feasible in the North-East-Flanders region of Belgium. Both the individual visits and the visits to the local quality groups were rated positively, and a majority of the general practitioners who returned the evaluation questionnaire wished to receive such visits in the future. The structure of the local quality groups implies limitations to the practical organisation of a visit, but requires less time investment than individual visits. PMID- 12761606 TI - Prediction of n-octanol/water partition coefficients for polychlorinated dibenzo p-dioxins using a general regression neural network. AB - A general regression neural network was used for the first time to study quantitative structure and property relationships of organic pollutants to correlate and predict n-octanol/water partition coefficients of polychlorinated dibenzo- p -dioxins from their topological molecular descriptors. In total, 42 polychlorinated dibenzo- p -dioxins and dibenzo- p -dioxins were available for this study-42 polychlorinated dibenzo- p -dioxins and dibenzo- p -dioxins in the training data set and 41 polychlorinated dibenzo- p -dioxins in the test data set. Partial least squares regression, back propagation network and general regression neural network models were trained using the training data set, and the accuracy of the models obtained were examined by the use of leave-one-out cross-validation. For prediction of the n-octanol/water partition coefficient, the best method is the general regression neural network. With the test data set, the correlation coefficient, root mean square error and mean absolute relative error for the general regression neural network model are 0.9276, 0.22 and 2.79%, respectively. For describing the structure of polychlorinated dibenzo- p dioxins, the topological molecular descriptors outperform the mobile order and disorder thermodynamic method. PMID- 12761607 TI - Varying and "atypical" indications for atypical antipsychotics. PMID- 12761605 TI - Paracetamol and metabolite pharmacokinetics in infants. AB - BACKGROUND: Data concerning metabolism of paracetamol in infants are scant. Previous studies have examined urinary metabolite recovery rates after a single dose of paracetamol in either neonates (<6 weeks) or children (3-9 years). There are no studies investigating infants. METHODS: Infants ( n=47) undergoing major craniofacial surgery were given paracetamol 19-45 mg/kg 6-, 8-, or 12-hourly as either elixir or suppository formulation for postoperative analgesia, after a loading dose of 33-59 mg/kg rectally during the operation. Serum was assayed for paracetamol concentration in 40 of these infants at 5, 8, 11, 14, 17 and 20 h postoperatively. Urine samples were collected every 3 h for 24 h in 15 of these infants. The clearances of paracetamol to glucuronide and sulphate metabolites as well as the urinary clearance of unmetabolised paracetamol were estimated using non-linear, mixed-effects models. RESULTS: Mean (+/-SD) age and weight of the patients were 11.8+/-2.5 months and 9.1+/-1.9 kg. Clearances of paracetamol to paracetamol-glucuronide (%CV) and to paracetamol-sulphate were 6.6 (11.5) l/h and 7.5 (11.5) l/h respectively, standardised to a 70-kg person using allometric "1/4 power" models. Glucuronide formation clearance, but not sulphate formation, was related to age and increased with age from a predicted value in a neonate of 2.73 l/h/70 kg to a mature value of 6.6 l/h/70 kg with a maturation half-life of 8.09 months. Urine clearance of paracetamol-glucuronide, paracetamol-sulphate and unchanged paracetamol (%CV) were, respectively, 2.65, 3.03 and 0.55 (28) l/h/70 kg. The urine clearance of unchanged paracetamol and metabolites was related to urine volume flow rate. Clearance attributable to pathways other than these measured in urine was not identifiable. The glucuronide/sulphate formation clearance ratio was 0.69 at 12 months of age. Sulphate metabolism contributed 50% towards paracetamol clearance. CONCLUSION: Glucuronide formation clearance increases with age in the infant age range but sulphate formation does not. Renal clearance of paracetamol and its metabolites increases with urine flow rate. This and other studies show that paracetamol metabolism to glucuronide appears to be similar in infants and children, but in adults is increased in comparison with children. Oxidative pathways were undetectable in this infant study and may explain, in part, the reduced incidence of hepatotoxicity in infants. PMID- 12761608 TI - Second-generation antipsychotics: discrepancies between licensed indications, evidence base and actual use. PMID- 12761609 TI - Hormone replacement therapy and risk for foot, distal forearm, proximal humerus, and pelvis fractures. AB - This case-control epidemiologic study examines the relationship between hormone replacement therapy (HRT) and risk for fractures of four sites among women 45 years of age and older. From October 1996 to May 2001, incident patients with distal forearm ( n=744), foot ( n=618), proximal humerus ( n=331), and pelvis ( n=109) fractures were recruited from five Kaiser Permanente Medical Centers in northern California. Controls ( n=1617) were selected at random from the same five medical centers over the same time period within strata defined by 5-year age group, gender, and white versus nonwhite or unknown recorded race/ethnicity. Trained interviewers collected information using a standardized questionnaire. Compared with postmenopausal women who never used HRT, postmenopausal women currently using HRT for at least 3 months had a decreased risk of fracture at the distal forearm (adjusted OR=0.55, 95% CI: 0.43, 0.72), proximal humerus (adjusted OR=0.51, 95% CI: 0.36, 0.74), and pelvis (adjusted OR=0.51, 95% CI: 0.27, 0.95), but not the foot (adjusted OR=1.05, 95% CI: 0.81, 1.35). Past use of HRT for more than 7 years also appeared to be associated with a reduced risk of fracture at the distal forearm and proximal humerus. The longer a woman went without HRT after her last menstrual period, the greater her risk of fracture in the distal forearm, proximal humerus, and pelvis. The findings of this study support long term, current use of HRT for the prevention of osteoporotic fractures, but other risks and benefits also need to be considered. PMID- 12761610 TI - Anterior cruciate ligament reconstruction in patients over the age of 50 years: 2 to 8-year follow-up. AB - We present medium-term results in 30 prospectively followed patients aged over 50 years at the time of ACL reconstruction (31 procedures); mean follow-up time was 46 months (range 24-95). Mean Lysholm score improved significantly from 63 pre operatively to 93 at final review Cincinnati score from 49 to 89. In IKDC score 25 knees (81%) were considered normal or nearly normal and 6 abnormal; there were no severely abnormal results. Mean Tegner activity scores improved from pre operatively from 3.7 to 5.2 at review. Mean side-to-side difference measured by the KT-1000 at maximal manual pressure was 2.7 mm; two knees had. a measured difference greater than 5 mm. The mean torque ratio for isokinetic flexion strength was 102% and for extension strength 94%. Poor results as determined by the three scoring systems were associated mainly with advanced articular degenerative changes (Outerbridge grade 3 or 4) seen at the time of reconstruction. Despite this all patients reported improvement in stability and overall function of the knee. Degenerative change itself was associated with increased time to surgery from injury. This study demonstrates that the ACL can be reliably reconstructed in patients over the age of 50 years with good symptomatic relief, restoration of function and return to sporting activity. PMID- 12761611 TI - The safety of the donor operation in living-donor liver transplantation: an analysis of 45 donors. AB - We retrospectively assessed the safety of the donor operation, based on parameters such as blood loss, blood transfusion, operation time, duration of hospitalization, and complications. Forty-five pediatric and adult recipients underwent living-donor liver transplantation (LDLTx) in Tohoku University Hospital from July 1991 to October 2000. Donor operations were classified into three groups. In the LS group, the graft was the lateral segment ( n=20); in the LL group, the graft was the left lobe without the middle hepatic vein ( n=16); and in the LLM group, the graft was the left lobe with the middle hepatic vein ( n=9). No significant differences were observed among the three groups regarding postoperative liver function or duration of hospitalization. In the LS group, the operation time was shorter and the requirement of autologous blood transfusion was significantly lower than in the other two groups. Most complications following retrieval of the graft were minor. Safety is guaranteed when the left lobe or the left lateral segment is used for LDLTx, but meticulous management of the operation is required to prevent complications. PMID- 12761613 TI - Treatment with recombinant human activated protein C obviates additional anticoagulation during continuous venovenous hemofiltration in patients with severe sepsis. PMID- 12761614 TI - Combined antithrombin and protein C supplementation in meningococcal purpura fulminans: a pharmacokinetic study. AB - PURPOSE: To document in patients with meningococcal purpura fulminans (PF), the effects of a combined supplementation with antithrombin (AT) and protein C (PC) plasma concentrates and to estimate the pharmacokinetics and dose requirements of each inhibitor. DESIGN: Retrospective study of 15 patients. SETTING. One paediatric and one adult ICU in a university hospital. INTERVENTIONS: In addition to standard intensive care, all patients received a 100 IU/kg loading dose of AT and PC concentrates, followed by a continuous infusion (AT: 100-150 IU.kg.day; PC: 100 IU.kg.day in adults, and 400 IU/kg in infants). MEASUREMENTS: Clinical data, coagulation, and fibrinolysis parameters, AT and PC activities, and free protein S (PS) levels were sequentially measured. Restitution ratio, median increment after supplementation, and half-life of clearance from plasma were calculated for the two plasma substitutes. RESULTS. At admission, all patients had a severe decrease in AT, PC, and PS levels. The supplementation regimen induced a substantial increase in AT and PC activities, peaking at H18 and H48, respectively. The supplementation procedure did not modify free PS levels. The median values of AT and PC restitution ratio, increment in plasma activity observed after 100 IU/kg concentrate, and apparent half-life of clearance from plasma were 0.85 U.ml.U.kg and 0.59 U.ml.U.kg, 23% and 21%, 16 h and 6 h, respectively. CONCLUSION: If AT and PC concentrates are to be given in fulminant meningococcemia, the doses of supplementation should be at least 150 IU/kg AT and 250 IU/kg PC as loading dose and 150 IU/kg AT and 200 IU/kg PC as daily maintenance therapy. Taking into account the individual variability in inhibitor deficiency and restitution ratio, repeated measurements of plasma levels are mandatory to obtain a patient-based adjustment of the supplementation. PMID- 12761615 TI - A novel citrate anticoagulation regimen for continuous venovenous hemodiafiltration. AB - OBJECTIVE: Validation of a novel citrate anticoagulation regimen for continuous venovenous hemodiafiltration (CVVHDF). DESIGN AND SETTING: Prospective cohort trial in medicosurgical intensive care units of two university-affiliated teaching institutions. PATIENTS: Participants were patients at high risk for bleeding, with renal failure requiring CVVHDF without heparin. Fourteen patients completed the study. INTERVENTION: A convection-based citrate anticoagulation CVVHDF regimen using an isotonic replacement fluid containing citrate administered in predilution. A neutralizing solution of calcium chloride and magnesium sulfate was infused at the end of the circuit. Blood flow rate was set and kept at 125 ml/min, and the flow rate of the replacement fluid was initiated at 1250 ml/h and adjusted thereafter according to the monitoring of blood activated coagulation time (ACT), with a target between 180 to 220 s. MEASUREMENTS AND RESULTS: The average filter time-life was 44 h. Thrombosis of the proximal portion of the circuit (which was not anticoagulated) was the main reason for technique failure. A mean urea clearance of 21 ml/min was obtained. Electrolytes and acid-base balance were both well maintained. Six percent (16/287) of Ca(i) readings less than 0.3 mmol/l were associated with very high ACT levels (>300 s). CONCLUSIONS: This regimen is shown to be safe, efficacious, and convenient. Citrate anticoagulation should be monitored using postfilter ACT and/or ionized calcium with respective targets of 200-250 s or 0.3-0.4 mmol/l. PMID- 12761616 TI - Abdominal compartment syndrome related to noninvasive ventilation. AB - OBJECTIVE: To study the effects of noninvasive positive pressure ventilation (NIPPV) on intra-abdominal pressure. DESIGN AND SETTING: Single case report from a tertiary teaching hospital. PATIENTS AND METHODS: A 65-year-old man who experienced a sudden respiratory and cardiovascular collapse during NIPPV. This was caused by gastric overdistension due to aerophagia followed by raised intra abdominal pressure leading to intra-abdominal hypertension and abdominal compartment syndrome. RESULTS: The respiratory and cardiovascular problems resolved immediately after the introduction of a nasogastric tube. This resulted in normalization of IAP. CONCLUSIONS: This is the first case reported of an abdominal compartment syndrome related to NIPPV. Clinicians should be aware of this possible complication while using NIPPV. PMID- 12761618 TI - Microalbuminuria: timing is everything! PMID- 12761617 TI - Norepinephrine for hypotensive vasodilatation after cardiac surgery: impact on renal function. AB - OBJECTIVES: Norepinephrine use in patients after cardiac surgery is controversial because of the fear that norepinephrine might decrease kidney function through regional vasoconstriction. Accordingly, we studied the renal effects of norepinephrine use for hypotensive vasodilatation after cardiac surgery. DESIGN AND SETTING: Retrospective controlled study in the cardiothoracic ICU of tertiary hospital. PATIENTS. 100 cardiac surgery patients with post-operative hypotensive vasodilatation and 100 control cardiac surgery patients. INTERVENTION: Treatment of hypotension (MAP<70 mmHg) with continuous norepinephrine infusion. MEASUREMENTS AND RESULTS: We collected data on demographic and surgical characteristics, haemodynamics, serum creatinine and mortality. Just after surgery the norepinephrine group had a significantly higher mean central venous pressure, lower mean arterial pressure, and lower systemic vascular resistance index with a similarly elevated mean cardiac index. Despite norepinephrine administration at a mean peak dose of 7.3+/-6.4 micro g/min the mean post operative change in creatinine was not different between two groups on days 0, 2 or 4 after surgery. CONCLUSIONS: Norepinephrine does not increase post-operative serum creatinine concentrations in patients with hypotensive vasodilatation after cardiac surgery. Concerns related to its potential adverse effects on the kidney function in this setting appear unjustified. PMID- 12761620 TI - Monte Carlo simulations on marker grouping and ordering. AB - Four global algorithms, maximum likelihood (ML), sum of adjacent LOD score (SALOD), sum of adjacent recombinant fractions (SARF) and product of adjacent recombinant fraction (PARF), and one approximation algorithm, seriation (SER), were used to compare the marker ordering efficiencies for correctly given linkage groups based on doubled haploid (DH) populations. The Monte Carlo simulation results indicated the marker ordering powers for the five methods were almost identical. High correlation coefficients were greater than 0.99 between grouping power and ordering power, indicating that all these methods for marker ordering were reliable. Therefore, the main problem for linkage analysis was how to improve the grouping power. Since the SER approach provided the advantage of speed without losing ordering power, this approach was used for detailed simulations. For more generality, multiple linkage groups were employed, and population size, linkage cutoff criterion, marker spacing pattern (even or uneven), and marker spacing distance (close or loose) were considered for obtaining acceptable grouping powers. Simulation results indicated that the grouping power was related to population size, marker spacing distance, and cutoff criterion. Generally, a large population size provided higher grouping power than small population size, and closely linked markers provided higher grouping power than loosely linked markers. The cutoff criterion range for achieving acceptable grouping power and ordering power differed for varying cases; however, combining all situations in this study, a cutoff criterion ranging from 50 cM to 60 cM was recommended for achieving acceptable grouping power and ordering power for different cases. PMID- 12761621 TI - Molecular differentiation and diversity among the California red oaks (Fagaceae; Quercus section Lobatae). AB - A recent epidemic of Phytopthora (Sudden Oak Death) in coastal woodlands of California is causing severe mortality in some oak species belonging to the red oak (Lobatae) group. To predict the risks of spread of this disease, an understanding of the relationships among California's red oak species and of their population genetic structure is needed. We focus here on relationships among the four species of red oak. Whereas morphological distinction of Quercus wislizeni and Quercus parvula can pose problems, Quercus kelloggii and Quercus agrifolia in pure forms are easily distinguishable from one another and from Q. wislizeni and Q. parvula in the field. However, hybrids among all species combinations are known to occur in nature and these can confound data from ecological studies. Our results revealed greatest differentiation of the deciduous Q. kelloggii, with only weak AFLP fragment differentiation of the three remaining evergreen species. The molecular data suggest a closer affinity of Q. agrifolia with Q. wislizeni and Q. parvula contrary to earlier suggestions that its origins are likely to have been with northern deciduous oaks probably through a common ancestor with Q. kelloggii. Interior and coastal populations of Q. wislizeni separated in dendrograms based on phenetic and genetic distances suggesting probable isolation in different glacial refugia. The position of Q. parvula remains ambiguous, having a closer affinity with interior populations of Q. wislizeni and with Q. agrifolia, than with coastal populations of Q. wislizeni. Mean population differentiation in Q. wislizeni was 0.18, which is somewhat higher than the average for other oak species, suggesting that range fragmentation has occurred in the past, resulting in a metapopulation structure. Our results provide evidence that introgression among these species may be causing reticulation, further confounding species separation. Whereas Phytopthora has been reported on Q. agrifolia, Q. parvula and Q. kelloggii, it has not yet been detected in natural populations of Q. wislizeni. The species relationships that our molecular data show suggest that this is more likely a result of escape due to ecological tolerances than to genetic differences. PMID- 12761622 TI - Detection of alternatively spliced EMR2 mRNAs in colorectal tumor cell lines but rare expression of the molecule in colorectal adenocarcinomas. AB - EMR2 and CD97, members of the epidermal growth factor (EGF)-TM7 family, show a very high homology. CD97, whose expression is closely related to clinical tumor stage in colorectal carcinomas, potentially functions as an adhesion molecule. Nothing is known about the presence of EMR2 in these tumors. We systematically examined the expression of EMR2 in colorectal carcinoma cell lines and adenocarcinomas. Of 18 cell lines, 10 were only slightly positive for EMR2 according to flow cytometry. Various EMR2 splice variants, including a new isoform, have been detected at the mRNA level. EMR2 expression did not correlate with in vitro migration or invasion capacity of the cell lines. Normal colorectal epithelial cells were EMR2 negative. In contrast to CD97, which is found in most colorectal adenocarcinomas, only 8 of 81 of these tumors expressed EMR2. No correlation was found between EMR2 expression and clinicopathological parameters of the tumors. In summary, a significant but low number of colorectal carcinomas are positive for EMR2, indicating different roles for this molecule and CD97 in these tumors. PMID- 12761624 TI - Stimulus-response compatibility between stimulated eye and response location: implications for attentional accounts of the Simon effect. AB - One influential theory of the Simon effect, the attention-shift hypothesis, states that attention movements are the origin of spatial stimulus codes. According to this hypothesis, stimulus-response compatibility effects should be absent when attention shifts are prevented. To test this prediction, we used monocular patches of color that required left or right key-press responses. About half of the subjects could discriminate which eye was stimulated (in a subsequent task), and showed strong spatial compatibility effects between the stimulated eye and the response location. The other half of the subjects could not make a utrocular discrimination (i.e., they could not judge which eye had received monocular stimulation), but the pattern of results was the same: the fastest reaction times were observed when the stimulated eye corresponded spatially to the required response (i.e., a Simon effect). Since the subjects presumably did not move their attention (from the subject's point of view, the stimuli were presented centrally), our results indicate that spatial codes can be produced in the absence of attention shifts. These results also show that utrocular discrimination can be assessed via indirect measures that are much more sensitive than explicit measures. PMID- 12761623 TI - Vascular endothelial cell participation in formation of lymphoepithelial lesions (epi-myoepithelial islands) in lymphoepithelial sialadenitis (benign lymphoepithelial lesion). AB - Lymphoepithelial lesions (LELs, or epi-myoepithelial islands) in lymphoepithelial sialadenitis (LESA, or benign lymphoepithelial lesion) of the salivary gland are known to be mainly composed of duct epithelial cells. However, other constituent cells are poorly characterized. Formalin-fixed paraffin sections obtained from six surgical specimens of LESA were examined using immunohistochemistry for cytoskeletal proteins, inflammatory cells, vascular endothelial cells, and extracellular matrix (ECM) molecules as well as by in situ hybridization for ECM molecules. In addition to keratin-immunopositive (+) duct-like epithelial cells, there were CD31/CD34+ vascular endothelial cells-which were either scattered in a singular fashion, in formed sheets, or in tubular structures-, CD20+ B lymphocytes, CD45RO+ T lymphocytes, and CD68 macrophages in the LELs. ECM molecules, such as heparan sulfate proteoglycan and tenascin, were immunolocalized in hyaline materials in the LELs. Their mRNAs were demonstrated mainly in endothelial cells and, to a lesser extent, in lympho-monocytic cells around hyaline materials, but were not as evident in epithelial constituent cells of LELs. The results indicate that endothelial cells as well as inflammatory cells are important constituents of the LELs, and the hyaline ECM cores mainly result from the intra-LEL angiogenesis by endothelial cells with the assistance of inflammatory cells. This intra-LEL vasculature seems to support regeneration and proliferation of salivary epithelial remnant cells. PMID- 12761625 TI - Role of constitutively expressed heme oxygenase-2 in the regulation of guinea pig coronary artery tone. AB - Carbon monoxide (CO) is well known as a relaxing substance in the vasculature, where it is released during the heme oxygenase (HO) reaction. Little is known about the tissue-specific targets of CO in smooth muscles. To date the functional role of CO in the coronary artery remains unclear. The expression of HO-2, the constitutive isoform of HO, but not of HO-1 (inducible HO isoform) was demonstrated by immunohistochemical reaction. Contractile studies, performed under isometrical conditions, showed that CO, as well as hemin (given as a substrate for HO), relax de-endothelized coronary smooth muscle after the blockade of neuronal transmission. The action of hemin was antagonized by preliminary treatment of the vessel with SnPPIX--a competitive inhibitor of HO. The relaxatory effects of hemin were abolished in the presence of guanylyl cyclase or protein kinase G antagonists. Patch-clamp studies revealed that hemin caused activation of iberiotoxin-blockable K outward current (I(K)) via guanylyl cyclase and protein-kinase-G-dependent mechanisms. This activation coincided with hyperpolarization of the plasma membrane of single coronary smooth muscle cells by 8+/-3 mV, which was prevented by preliminary exposure of cells to 10 microM SnPPIX. The I(K)-augmenting effect of hemin was not affected by pretreatment of cells with cyclopiazonic acid and/or ryanodine, blockers of phospholipase C or heparin (applied via pipette), but was not observed when ATP was omitted from the dialyzing solution, or in the presence of Na-free, ATP-containing pipette solution. The omission of Ca(2+) from the bath or the replacement of Na with Li in both pipette and bath media also prevented the I(K)-activating effect of hemin. These results suggest that the constitutive HO-2 in coronary artery smooth muscle cells plays role in the modulation of tone. At the level of smooth muscle cells CO and its precursor hemin may cause hyperpolarization of the plasma membrane by activation of iberiotoxin-sensitive I(K) presumably via PKG-dependent activation of the Na/Ca exchanger. This activation is thought to increase the submembrane Ca(2+) concentration in the vicinity of large-conductance, Ca(2+) sensitive K channels, thus causing voltage-dependent inhibition of Ca(2+) entry and subsequent relaxation of the vessel. PMID- 12761626 TI - Skin protection and percutaneous absorption of chemical hazards. AB - Hazardous substances that penetrate the skin barrier may induce dermal irritation, inflammation or sensitisation, as well as systemically toxic effects. Air threshold limits are insufficient to prevent adverse health effects in the case of contact with substances with a high dermal absorption potential. Biological monitoring is a useful tool to quantify internal exposure of an individual and allows an assessment to be made of the dermal absorption pathway. Additionally, the efficacy of protective equipment can be proved. Standard gloves are ineffective to decrease dermal uptake. Various skin-care products include emulgators, which are used as penetration enhancers in transdermal therapeutic drugs. Hence, specific skin-care products potentially enhance dermal absorption of certain chemicals. On the other hand, the best protection from dermal absorption is the physiological skin barrier-the stratum corneum. Skin-care products that accelerate regeneration of the stratum corneum are obliged to have positive effects to prevent dermal absorption. With regard to the increasing number of occupational skin diseases and the large quantity of skin-care products that are used at workplaces it is essential that the mechanism of protection and efficacy be assessed. In the future, we require evidence-based prevention or evidence-based medicine that includes evidence-based prevention. PMID- 12761627 TI - Molecular mechanisms of Bartter syndrome caused by mutations in the BSND gene. AB - Barttin, a gene product of BSND, was identified as a fourth gene responsible for Bartter syndrome. The co-expression of barttin with CLC-K chloride channels has been demonstrated to dramatically induce the expression of CLC-K current. However, it remains unknown how barttin interacts with CLC-K channels in mammalian cells and how the mutations of barttin lead to Bartter syndrome. In an attempt to clarify the effect of barttin expression on CLC-K2 cellular localization, we examined the expression of the CLC-K2 chloride channel and barttin, solely and in combination, in transient and stable expression systems in mammalian cells. In addition, we generated several stable cell lines expressing mutant barttins to clarify the consequence of the previously reported barttin mutations in Bartter syndrome. In immunocytochemistry, CLC-K2 was retained in the Golgi in the absence of barttin expression, but delivered to the plasma membrane when barttin was present. Barttin was coprecipitated with CLC-K2, suggesting a protein-protein interaction. Disease-causing mutant barttins, especially R8L, were retained intracellularly, but their binding ability to CLC-K2 was preserved. This led to a retention of CLC-K2 in intracellular organelles with barttin, and a loss of plasma membrane localization. The stability of the CLC-K2 protein was also markedly increased by coexpression with barttin. These results clarified that barttin determined cellular localization of CLC-K2 by protein-protein interaction. Thus, the mislocalization of CLC-K2 was identified as the molecular pathogenesis of Bartter syndrome by mutant barttins. PMID- 12761628 TI - Therapeutic strategies for Parkinson's disease based on data derived from genetic research. AB - Following the identification of mutations in alpha-synuclein as the cause of some rare forms of familial Parkinson's disease (PD), genetic research has uncovered numerous gene loci of PD. Meanwhile, several neurodegenerative diseases have been shown to accumulate a-synuclein in neuronal and glial cells summarizing this group of diseases as synucleinopathies. All currently known gene defects causing PD alter the ubiquitin-proteasomal pathway of protein degradation. Identification of these disease mutations allows studying the functional consequences which lead to cellular dysfunction and cell death in cell culture and transgenic animal models, to identify therapeutic targets and to test potential protective strategies in these models. PMID- 12761629 TI - Differential diagnosis of parkinsonian syndromes: dynamics of time courses are essential. AB - Differential diagnosis of parkinsonian syndromes encloses idiopathic Parkinson's disease, other primary neurodegenerative diseases (atypical parkinsonism), symptomatic cases, pseudoparkinsonism and inherited forms. Clinical diagnostic criteria are not safe when applied only at disease onset. Diagnostic accuracy can be intensified by recognition of distinct patterns of signs and symptoms and by focussing attention on the time course of the disease. PMID- 12761630 TI - Functional imaging in Parkinson's disease: activation studies with PET, fMRI and SPECT. AB - Activation studies with positron emission tomography (PET) and functional magnetic resonance imaging (fMRI) represent a powerful tool to study the functional anatomy of Parkinson's disease (PD). Activation studies offer the opportunity to study regional cerebral function in man in vivo under different conditions with the analysis of task specific changes in regional cerebral blood flow (rCBF) with PET or in the blood oxygenation level dependent (BOLD) effect with fMRI. The combination of PET and deep brain stimulation is particularly attractive to study the effects of discrete perturbations at different target structures throughout the basal ganglia-thalamocortical circuitries. The use of rCBF PET and fMRI to study the pathophysiology of PD in the motor and sensory system and mechanisms of dopaminergic therapy as well as surgical interventions will be reviewed. PMID- 12761631 TI - Does ultrasound disclose a vulnerability factor for Parkinson's disease? AB - Transcranial ultrasound is a new tool allowing the detection of abnormalities in the echomorphology of the substantia nigra (SN) in patients with Parkinson's disease (PD). Several lines of evidence suggest that the changes in the echo pattern represent a risk factor as: i) the majority of PD patients exhibit this echo-feature, ii) the presence of such changes in healthy controls is related to a reduced (18)F-Dopa-uptake and clinical signs of nigrostriatal dysfunction. The reason for the change of echogenicity is suggested to be an increased iron content in the substantia nigra causing oxidative stress and neuronal cell damage. This hypothesis of changes in SN echomorphology reflecting a risk factor of PD has to be proved in longitudinal studies. PMID- 12761632 TI - Autonomic dysfunctions in idiopathic Parkinson's disease. AB - Most patients with Parkinson's disease experience autonomic dysfunction at some point in the course of their disease. In contrast to autonomic dysregulation in multisystem atrophy, they are less severe, and they frequently cause troublesome symptoms only in advanced stages of the disease. The quality of life is nevertheless substantially restricted. Cardiovascular, gastrointestinal and urogenital autonomic dysfunction is predominant. Appropriate diagnosis and treatment can greatly benefit the patient. An interdisciplinary approach is desirable in most cases. PMID- 12761633 TI - The relevance of preclinical studies for the treatment of Parkinson's disease. AB - An essential element of pharmaceutical development, defined as the period between the discovery of a new agent and its market release, is provided by the "preclinical studies". They consist of the in vitro and in vivo studies performed before examination of the agent in human subjects. Regulatory authorities prescribe specific requirements regarding the nature and number of preclinical studies. In the present paper, we discuss the relevance of these studies for the treatment of Parkinson's disease (PD) on the basis of three examples: the L-DOPA ( L-3,4-dihydroxyphenylalanine, levodopa) story; the development of selegiline as a palliative and neuroprotective drug; and the safety concerns regarding tolcapone, an inhibitor of central and peripheral catechol-O-methyltransferase (COMT). PMID- 12761634 TI - Inflammation in Parkinson's disease. AB - Several studies of Parkinson's disease (PD) patients and experimental models of PD indicate the presence of an inflammatory process in PD. Although the primary cellular mechanisms remain to be clarified, activation of resident microglia appears to aggravate or even maintain the disease process in PD. Modulation of inflammatory mechanisms could provide a new neuroprotective therapy in PD. PMID- 12761635 TI - Diffuse Lewy body disease - a clinical syndrome or a disease entity? AB - Most clinicians and researchers still accept diffuse Lewy body disease (DLBD) as a clinicopathological entity. Dementia with fluctuating cognitive deficits, a parkinsonian syndrome, and visual hallucinations are the core symptoms of this proposed disease entity. From a neuropathological point of view, many examples of patients with progressive dementia showing evidence of extensive Lewy body formation in the cerebral cortex together with the occurrence of Lewy bodies in substantia nigra and locus coeruleus have been identified. Confusingly, a large majority of cases showing typical features of DLBD also present with an Alzheimer pathology in the hippocampus and cerebral cortex. It is far from clear that DLBD represents a specific disease entity rather an intermediate variant between Alzheimer disease and idiopathic parkinsonian syndromes. Nevertheless, from a clinical point of view it may be of importance to characterize patients with a symptomatology of DLBD because important management issues such as avoidance of severe neuroleptic sensitivity reactions, dopaminergic antiparkinsonian treatment and a beneficial response to cholinesterase inhibitors can be applied. PMID- 12761636 TI - Deep brain stimulation of the subthalamic nucleus for Parkinson's disease: a therapy approaching evidence-based standards. AB - DBS of the STN is one of the most promising new therapies for the treatment of PD. However - like many other therapies for PD - the present stage of the scientific assessment does not yet suffice the rigid criteria of evidence-based medicine. Further studies should specifically address the questions of efficacy and side effects as well as the impact on quality of life. PMID- 12761637 TI - Deep brain stimulation in dystonia. AB - Renewed interest in stereotaxy for dystonia followed the introduction of deep brain stimulation (DBS) in Parkinson's disease and essential tremor in the 1990s. DBS evolved from ablative surgery, which was applied with varying results in the 1950s in patients with movement disorders such as Parkinson's disease, essential tremor and dystonia. The present review summarizes the current knowledge on clinical aspects of DBS in dystonia (Dec. 2002). Excellent results have been achieved in dystonic patients carrying a mutation in the DYT1 gene with improvements up to 90 %. Similar results may also be obtained in patients with idiopathic generalized dystonia, myoclonus-dystonia syndrome, and tardive dystonia. Substantial improvement has been observed in patients with focal dystonia (for instance cervical dystonia). Patients with secondary dystonia often display a lesser and more variable degree of improvement. Long-term studies are warranted to assess both motor and neuropsychological sequelae of DBS in dystonia. Furthermore, the optimal target for different dystonic disorders remains to be determined, although the globus pallidus internus has currently emerged as the most promising target for dystonia. PMID- 12761638 TI - Midline pyogenic granuloma of the tongue: two case studies. AB - Characteristically, pyogenic granuloma of the tongue is more common on the lateral side of the tongue. The reasons for this are unclear, but may be related to trauma from adjacent teeth or dentures. This article was prompted by the presentation of two patients with lesions in the midline of the anterior surface of the dorsum of the tongue. Both lesions were successfully removed surgically, and histology revealed them to be pyogenic granulomas. There has been no evidence of recurrence in the 6th month of the follow-up period. PMID- 12761639 TI - Investigation of radiosialadenitis during fractioned irradiation: sialoscintigraphical and histomorphological findings in rats. AB - Xerostomia is the most debilitating side effect induced by irradiation of head and neck tumours and is caused by irradiation damage to the salivary glands. The aim of this study was to correlate structural histomorphological damages and sialoscintigraphical findings during fractioned radiotherapy. The head and neck area of 27 WAG/RijH rats was irradiated with 60Co-gamma rays (60 Gy/30f 6 weeks). To evaluate salivary gland function, a port system was implanted, and 99mTc pertechnetate was applied at different stages of irradiation (0, 16, 30, 46, 60 and 6 months post-irradiation). In the course of treatment the parotid glands were examined histopathologically. Rat salivary glands developed a dose-dependent radiosialadenitis. After a dose of 16 Gy an intra- and extra-cellular oedema developed in the salivary glands. Progressive vacuolisation (30 Gy) developed into lipomatosis (46 Gy) and necrotic changes (60 Gy) in the parotid glands. Six months after irradiation treatment, the chronic histomorphological damages corresponded to stage II according to Seifert. The corresponding loss in gland function investigated by measurement of the 99mTc-pertechnetate uptake of the salivary glands was 13% (16 Gy), 26% (30 Gy), 57% (46 Gy), 75% (60 Gy) and 66.5% (6 months post-irradiation). The presented animal model is suitable to demonstrate the correlation of histomorphological and sialoscintigraphical findings. PMID- 12761640 TI - Hip revision with impacted morselized allografts: unrestricted weight-bearing and restricted weight-bearing have similar effect on migration. A radiostereometry analysis. AB - BACKGROUND: No consensus exists as to whether to let patients bear weight or not after revision of a loosened hip prosthesis using morselized and impacted allograft bone and cement. In the original description of the method, the patients were advised not to bear weight for 3 months postoperatively. Theoretically, bone graft remodeling is enhanced by mechanical load, but an increased migration of the prosthesis within the graft can also be anticipated. In addition, practicing restricted weight-bearing is cumbersome for the patients. METHODS: The present study was initiated to evaluate the migration, as measured by radiostereometry analysis (RSA), in patients mobilized with unrestricted weight-bearing after a hip revision using the X-change Revision Instruments System. This group was compared with a previous series with restricted weight bearing for the initial 3 months postoperatively. Seven stems and 12 sockets were evaluated by RSA over 2 years. Only patients without intraoperative complications were mobilized with unrestricted weight-bearing. RESULTS: All 7 stems migrated in the distal direction (median 2.6 mm; range 0.8-16.5 mm) and in the posterior direction (median 2.3 mm; range 0.7-22.1 mm). Eleven of the 12 sockets migrated in the proximal direction (median 2.5 mm; range 0.2-8.1 mm). The migration rate decreased gradually in all directions, but 3 stems and 5 sockets still migrated between the 1.5- and 2-year follow-ups. There were no significant differences in migration in any direction for either the stems or the sockets compared to the group with restricted weight-bearing. CONCLUSION: No increased migration occurred in the group free to bear weight as compared to restricted weight-bearing. We shall continue to allow unrestricted weight-bearing in cases where the femoral bone feels competent to withstand the initial load. It simplifies the postoperative mobilization, and we speculate that it might increase the remodeling of the graft. PMID- 12761641 TI - Vascular endothelial growth factor in inflammatory bowel disease. AB - BACKGROUND AND AIMS: Angiogenesis is an important component of tissue regeneration. As Inflammatory bowel disease (IBD) involves inflammation, ulceration, and regeneration of the intestinal mucosa, angiogenesis may be an integral part of IBD pathology. This study investigated the role of vascular endothelial growth factor in IBD. PATIENTS AND METHODS: The VEGF plasma (pVEGF) and serum (sVEGF) levels were assessed in patients with ulcerative colitis (UC; n=50) or Crohn's disease (CD; n=44) and in healthy controls (n=23). The immunohistochemical expression of VEGF was also assessed in surgical material from 11 patients with active IBD. RESULTS: Overall the sVEGF levels ranged from 30-899 pg/ml (median 200 pg/ml) and were significantly higher than the pVEGF levels (range 20-80 pg/ml, median 30 pg/ml). pVEGF levels were significantly lower in patients with active and quiescent CD than in healthy controls. Despite the lower pVEGF levels noted also for patients with UC, the difference was not significant. sVEGF levels were also reduced in patients with IBD, but the difference was not significant. No association of pEGF/sVEGF with beta thromboglobulin and platelet factor 4 levels (markers of platelet activation) was noted. On immunohistochemistry VEGF was not expressed in the inflammatory component (lymphocytes and macrophages), the fibroblasts, or the muscular layer of the intestinal wall. The intestinal epithelium was negative in CD, while a cytoplasmic reactivity was noted in UC and normal controls. CONCLUSION: As VEGF is a vascular and epithelial cell survival factor, the defective VEGF response ability, confirmed here for patients with CD, may be a key element in the pathology of the disease. The pathology of UC, however, seems not to be VEGF dependent. PMID- 12761642 TI - Dynamic transperineal ultrasound vs. defecography in patients with evacuatory difficulty: a pilot study. AB - BACKGROUND AND AIMS: Defecating proctography has been traditionally used to assess patients with evacuatory dysfunction. More recently, dynamic transperineal ultrasound has been described, defining the interaction between the infralevator viscera and the pelvic floor at rest and during straining. This study compared qualitative diagnosis and quantitative measurement obtained by defecography and dynamic transperineal ultrasonography in patients with evacuatory difficulty. PATIENTS AND METHODS: Thirty-three women were examined using both techniques with both examiners blinded to the results of the other method. Quantitative measurement was made of rectocele depth, anorectal angle (at rest and during maximal straining) and anorectal junction position at rest and movement during straining. RESULTS: There was good agreement for the diagnoses of rectocele, rectoanal intususseption, and rectal prolapse. Dynamic transperineal ultrasound was more likely than defecography to make multiple diagnoses or to diagnose an enterocele when a rectocele was present. There was no difference noted between the two techniques for the measurement of anorectal angle at rest, anorectal junction position at rest, or anorectal junction movement during straining. The mean anorectal angle during straining was 123.3+/-4.3 degrees as measured by defecography and 116.4+/-3.3 degrees as measured by dynamic transperineal ultrasound, nearly reaching statistical significance. CONCLUSION: Dynamic transperineal ultrasound is a simple and accurate technique for assessment of the pelvic floor and soft-tissues in patients with evacuatory dysfunction. PMID- 12761643 TI - Pilocytic astrocytomas with leptomeningeal dissemination: biological behavior, clinical course, and therapeutical options. AB - BACKGROUND: The aim of this case-based update is to discuss the biological characteristics, clinical course, and optimal treatment of pediatric patients harboring pilocytic astrocytomas (PA) with leptomeningeal dissemination (LMD). PAs are well-recognized benign pediatric tumors of the central nervous system (CNS). Despite a favorable outcome in general, in rare cases LMD might occur and contribute to poor clinical course. Localization of the PA in the chiasmo hypothalamic region seems especially predisposed to LMD. In addition, specific histological features might constitute the potential for subarachnoid metastases. The optimal treatment is still under discussion but studies suggest that chemotherapeutical treatment should be used as first choice therapy in order to avoid or delay radiotherapy. CASE REPORTS AND DISCUSSION: Two cases of subtotally resected chiasmo-hypothalamic PAs with synchronous LMD but with different rates of progression are described. The literature is reviewed and compared with our findings. PMID- 12761645 TI - Fine control of call frequency by horseshoe bats. AB - The auditory system of horseshoe bats is narrowly tuned to the sound of their own echoes. During flight these bats continuously adjust the frequency of their echolocation calls to compensate for Doppler-effects in the returning echo. Horseshoe bats can accurately compensate for changes in echo frequency up to 5 kHz, but they do so through a sequence of small, temporally-independent, step changes in call frequency. The relationship between an echo's frequency and its subsequent impact on the frequency of the very next call is fundamental to how Doppler-shift compensation behavior works. We analyzed how horseshoe bats control call frequency by measuring the changes occurring between many successive pairs of calls during Doppler-shift compensation and relating the magnitude of these changes to the frequency of each intervening echo. The results indicate that Doppler-shift compensation is mediated by a pair of (echo)frequency-specific sigmoidal functions characterized by a threshold, a slope, and an upper limit to the maximum change in frequency that may occur between successive calls. The exact values of these parameters necessarily reflect properties of the underlying neural circuitry of Doppler-shift compensation and the motor control of vocalization, and provide insight into how neural feedback can accommodate the need for speed without sacrificing stability. PMID- 12761644 TI - Ependymoma. AB - INTRODUCTION: The management of a patient with an ependymoma is controversial. Although the necessity for a multi-disciplined approach is accepted, the exact roles for all disciplines are poorly defined. REVIEW: This review article examines the current status of histopathological and cytogenetic diagnosis, surgical, chemotherapeutic and radiotherapeutic treatment and future directions. PMID- 12761647 TI - The role of ultrasound in the diagnosis, management and evolutive prognosis of acute left-sided colonic diverticulitis: a review of 208 patients. AB - The aim of this study was to evaluate the role of ultrasound in the diagnosis and management of acute diverticulitis and its capacity to predict posterior complications in patients undergoing medical treatment. A review was made of the histories of 208 hospitalized patients (262 admissions) initially diagnosed with acute diverticulitis over a 5-year period. Ultrasound was performed in all patients upon first admission. Diverticulitis was retrospectively classified as either simple or complicated, the latter being defined by the presence of extraluminal air and/or abscesses. Diverticulitis was finally diagnosed in 203 patients. Ultrasound exhibited a sensitivity of 86% in 77 cases subjected to surgery, and of 94% in the global 203 patients (192 true-positive and 11 false negative findings). Of 34 patients with diverticulitis and emergency surgery, 10 had false-negative US exams. Twenty of the 73 cases (27%) with signs of complicated diverticulitis in the initial ultrasound study required emergency surgery, compared with only 4 of the 119 patients (3%) with US evidence of simple diverticulitis (p<0.001). Of the 169 patients with diverticulitis undergoing conservative management, 54 (32%) developed complications during follow-up. The patients under age 50 years with signs of complicated diverticulitis suffered more complications (65%) than the rest of groups (p<0.001). In subjects with recurrences (26%), these were either similar to or less than the first episode in 84% of the cases. The present study shows that ultrasound constitutes a feasible technique for diagnosing acute diverticulitis. The severity of diverticulitis according to US is statistically predictive of surgical risk during the acute phase. Severity is also related to the appearance of posterior complications in patients undergoing conservative management, although only in younger patients (<50 years). PMID- 12761649 TI - Veno-occlusive disease in a male patient with Marfan syndrome and common acute lymphoblastic leukemia during induction therapy. AB - Veno-occlusive disease (VOD) of the liver is characterized by jaundice, painful hepatomegaly, and retention of fluids. VOD is a severe complication in allogeneic stem cell or bone marrow transplantation. Additionally, the disease has been described in children suffering from nephroblastoma or rhabdomyosarcoma, treated with intense chemotherapy. Recently, VOD has been shown to be a complication in the treatment of myeloid leukemia with anti-CD33 linked to calicheamicin. We report the unusual case of a 21-year-old male patient with Marfan syndrome, diagnosed of acute lymphoblastic leukemia, who developed severe VOD during induction therapy after a single application of 2 mg vincristine. We speculate that coincidence of Marfan syndrome and application of induction chemotherapy might favor the disease in our patient. PMID- 12761648 TI - Biliary excretion of 17-(allylamino)-17-demethoxygeldanamycin (NSC 330507) and metabolites by Fischer 344 rats. AB - PURPOSE: 17-(Allylamino)-17-demethoxygeldanamycin (17AAG), an analogue of the benzoquinone ansamycin geldanamycin, has been extensively studied preclinically and is being evaluated clinically. Studies were performed to define the biliary excretion of 17AAG after i.v. delivery to rats, and to characterize the metabolites of 17AAG observed in rat bile. MATERIALS AND METHODS: In vivo studies were performed in bile-duct-cannulated Fischer 344 rats given a 10 mg/kg i.v. bolus dose of 17AAG. In vitro studies were performed with cloned human CYPs and microsomal epoxide hydrolase. Biliary excretion of 17AAG and metabolites was quantified by HPLC and followed for 4 h after drug delivery. 17AAG metabolites in bile and in in vitro reaction mixtures were identified with LC/MS/MS. RESULTS: By 15 min after i.v. delivery of 17AAG, bile contained at least 15 biotransformation products with absorbance spectra similar to that of 17AAG. Of these, metabolites eluting at 2.7, 2.9, and 8.6 min were present in sufficient concentrations to be quantified, although the lack of authentic standards resulted in their being expressed as 17AAG equivalents. Within the first 4 h after drug delivery, biliary excretion accounted for 28.9+/-6.1% of the 10-mg/kg 17AAG dose. 17AAG and 17 (amino)-17-demethoxygeldanamycin (17AG) accounted for 4.1+/-1.0% of the delivered dose, with 17AAG accounting for 2.0+/-0.5% and 17AG accounting for 2.1+/-0.5%. The metabolites eluting at 2.7, 2.9, and 8.6 min accounted for 10.6+/-2.0%, 9.8+/ 1.2%, and 1.0+/-0.2%, respectively, of the administered dose. LC/MS/MS analysis of bile demonstrated major metabolites with molecular weights of 545 and 619, corresponding to 17AG and the diol previously described as resulting from metabolism of 17AAG by CYP3A and microsomal epoxide hydrolase. Of the remaining proposed metabolites, ten had a mass and MS/MS spectrum consistent with mono oxygenated 17AAG metabolites. One of these metabolites has been identified as the epoxide previously described as resulting from CYP3A oxidation of the allyl double bond. Two other proposed metabolites had a mass and MS/MS spectrum consistent with demethylated 17AAG metabolites, and one had a mass and MS/MS spectrum consistent with a di-demethylated 17AAG metabolite. An analogous series of demethylated and oxidized metabolites was also observed for the 17AG metabolite. CONCLUSIONS: Biliary excretion of 17AAG represents a major route of elimination, although most of the material excreted is in the form of metabolites. Bile of rats dosed with 17AAG contained a number of metabolites not previously identified in the plasma or urine of mice treated with 17AAG, but analogous to metabolites described in bile of rats treated with 17 (dimethylaminoethylamino)-17-demethoxygeldanamycin (17DMAG, NSC 707545), another geldanamycin analogue undergoing preclinical evaluation in preparation for subsequent clinical trials. PMID- 12761650 TI - Primary non-Hodgkin's lymphoma of bone: three cases and a short review of the literature. AB - Primary non-Hodgkin's lymphoma of bone (PLB) is a rare entity. We present three new cases and a review of the literature. If PLB is suspected radiologically, the diagnosis must be confirmed by open biopsy. Histopathologically, PLB usually represents diffuse large B-cell lymphoma or lymphoma of the follicular center type. The extent of local disease manifestation, additional skeletal involvement, and the presence or absence of lymphadenopathy is assessed by radiological examination. Localized stages of the disease require involved-field radiotherapy (45-50 Gy) to the entire bone that is affected. Regional lymphatics must also be irradiated. Radiotherapy may be required immediately to prevent pathological fractures. In the few cases of more widespread skeletal or extraskeletal involvement, radiotherapy of the major site of manifestation may be followed by a "watch-and-wait" strategy until progression of the disease becomes apparent. In cases of intermediate or high-grade lymphoma of bone, combined radiochemotherapy is the treatment of choice for all stages. Six to eight cycles of chemotherapy (usually the CHOP regimen) are recommended for remission induction. This is followed by involved-field radiotherapy with 45-50 Gy. High-dose chemotherapy with autologous stem cell support is an option if there is no satisfactory response to conventional chemotherapy, or if early relapse occurs. PMID- 12761651 TI - Locked intramedullary nailing for difficult nonunions of the humeral diaphysis. AB - Antegrade intramedullary nailing and bone grafting was carried out for 27 patients with resistant atrophic nonunion of the humeral diaphysis. The initial fracture was open in 12 cases and closed in 15. There were ten proximal humeral fractures, 13 mid-shaft fractures and four distal humeral shaft fractures. Most had previous attempts at internal fixation with bone grafting. Fifteen cases united, but 12 remained ununited necessitating further surgical treatment. The failures were all in the more complex cases. Lack of rigidity and compression may be the problem. PMID- 12761653 TI - Biosynthesis of medium-chain-length poly(hydroxyalkanoates) with altered composition by mutant hybrid PHA synthases. AB - Pseudomonas resinovorans harbors two isogenic poly(hydroxyalkanoates) (PHAs) synthase genes ( phaC1(Pre), phaC2(Pre)) responsible for the production of intracellular medium-chain-length (mcl-)PHAs. Sequence analysis showed that the putative gene-products of these genes contain a conserved alpha/beta-hydrolase fold in the carboxy-terminal half of the proteins. Hybrid genes pha7 and pha8 were constructed by exchanging the alpha/beta-hydrolase-fold coding portions of phaC1(Pre) and phaC2(Pre) at the 3' terminal. When grown with decanoate as carbon source, the pha7- or pha8-transformed Escherichia coli LS1298 produced PHAs containing 73-75% beta-hydroxydecanoate (beta-HD) and 25-27% beta hydroxyoctanoate (beta-HO). Deletion mutants, Delta pha7 and Delta pha8, were isolated during the PCR-based construction of pha7 and pha8, respectively. Cells harboring these mutants produced PHAs containing 55-60 mol% beta-HD and 40-45 mol% beta-HO. These results demonstrate the feasibility of generating active hybrid mcl-PHA synthase genes and their mutants with the potential of producing polymers having a varied repeat-unit composition. PMID- 12761655 TI - Concurrent disjoint and reciprocal classification by Cebus apella in seriation tasks: evidence for hierarchical organization. AB - We report the results of a 4-year-long study of capuchin monkeys ( Cebus apella ) on concurrent three-way classification and linear size seriation tasks using explicit ordering procedures, requiring subjects to select icons displayed on touch screens rather than manipulate and sort actual objects into groups. The results indicate that C. apella is competent to classify nine items concurrently, first into three disjoint classes where class exemplars are identical to one another, then into three reciprocal classes which share common exemplar (size) features. In the final phase we compare the relative efficiency of executive control under conditions where both hierarchical and/or linear organization can be utilized. Whilst this shows a superiority of categorical based size seriation for a nine item test set suggesting an adaptive advantage for hierarchical over linear organization, Cebus nevertheless achieved high levels of principled linear size seriation with sequence lengths not normally achieved by children below the age of six years. PMID- 12761654 TI - Cloning and sequence analyses of a 2,3-dihydroxybiphenyl 1,2-dioxygenase gene ( bphC) from Comamonas sp. SMN4 for phylogenetic and structural analysis. AB - A genomic library of biphenyl-degrading Comamonas sp. SMN4 for isolating fragments containing the 2,3-dihydroxybiphenyl 1,2-dioxygenase (23DBDO) gene was constructed. The smallest subclone (pNPX9) encoding 23DBDO activity was sequenced and analyzed. The C-terminal domain of 23DBDO from Comamonas sp. SMN4 had five catalytically essential residues and was more highly conserved than the N terminal domain. Phylogenetic and structural relationships of 23DBDO from Comamonas sp. SMN4 were analyzed. PMID- 12761656 TI - Development of interception of moving targets by chimpanzees (Pan troglodytes) in an automated task. AB - The experiments investigated how two adult captive chimpanzees learned to navigate in an automated interception task. They had to capture a visual target that moved predictably on a touch monitor. The aim of the study was to determine the learning stages that led to an efficient strategy of intercepting the target. The chimpanzees had prior training in moving a finger on a touch monitor and were exposed to the interception task without any explicit training. With a finger the subject could move a small "ball" at any speed on the screen toward a visual target that moved at a fixed speed either back and forth in a linear path or around the edge of the screen in a rectangular pattern. Initial ball and target locations varied from trial to trial. The subjects received a small fruit reinforcement when they hit the target with the ball. The speed of target movement was increased across training stages up to 38 cm/s. Learning progressed from merely chasing the target to intercepting the target by moving the ball to a point on the screen that coincided with arrival of the target at that point. Performance improvement consisted of reduction in redundancy of the movement path and reduction in the time to target interception. Analysis of the finger's movement path showed that the subjects anticipated the target's movement even before it began to move. Thus, the subjects learned to use the target's initial resting location at trial onset as a predictive signal for where the target would later be when it began moving. During probe trials, where the target unpredictably remained stationary throughout the trial, the subjects first moved the ball in anticipation of expected target movement and then corrected the movement to steer the ball to the resting target. Anticipatory ball movement in probe trials with novel ball and target locations (tested for one subject) showed generalized interception beyond the trained ball and target locations. The experiments illustrate in a laboratory setting the development of a highly complex and adaptive motor performance that resembles navigational skills seen in natural settings where predators intercept the path of moving prey. PMID- 12761657 TI - Refined localization of dominant intermediate Charcot-Marie-Tooth neuropathy and exclusion of seven known candidate genes in the region. AB - Charcot-Marie-Tooth (CMT) neuropathy is one of the most common hereditary disorders of the human peripheral nervous system. The CMT syndrome includes weakness and atrophy of distal muscles, high arched feet (pes cavus), depressed or absent deep tendon reflexes, and mild sensory loss. Dominant intermediate CMT (DI-CMT) neuropathy is a form of CMT with intermediate median motor nerve conduction velocities. We previously localized the DI-CMT locus to a 16.8-cM region on chromosome 19p12-p13.2. Extended haplotype analysis and clinical assessment of additional family members and a report of a second family linked to this locus has enabled us to narrow the candidate region to a 6-cM interval flanked by D19S558 and D19S432. Selection of positional candidate genes for screening was performed on the basis of neural expression and microarray analysis of Schwann cell differentiation in vivo. Seven candidate genes have been investigated. These include six genes localized in the original linkage interval and one in the newly refined region. They are excluded as a cause for DI-CMT neuropathy. PMID- 12761658 TI - Haplotype analysis of the ETM2 locus in familial essential tremor. AB - The objective of this study was to analyze a sample of unrelated individuals with autosomal dominant essential tremor (ET) for a genetic association with loci in a candidate region (ETM2) on chromosome 2p24.1 that harbors a disease gene for ET. ET is a common movement disorder that is genetically linked to ETM2 in four large families. It is unknown whether this candidate locus is associated with dominantly inherited ET in other individuals. Based on information from previous genetic linkage studies, a linkage disequilibrium study was designed to compare individuals with a family history of ET (n=45) with normal controls (n=70). Three unreported dinucleotide polymorphic loci (etm1240, etm1231, and etm1234) were identified on a physical map of the ETM2 interval in a region of no recombination. The study sample was tested for allele frequency differences by the CLUMP program and haplotypes were analyzed by the FASTEHPLUS program. The allele frequencies were significantly different between ET cases and the control samples for the loci etm1231 (P< or =0.0419) and etm1234 (P<0.0001). A haplotype formed by the loci etm1231 and etm1234 occurred with a frequency of 29% in cases (n=45) and 9% in a white newborn sample (P<0.0001, n=35). The haplotype was not found in normal individuals older than 60 years without tremor (P=0.0063, n=35). This study provides evidence that an ancestral haplotype on chromosome 2p24.1 segregates with the ET disease phenotype in individuals with a family history of the disorder and will facilitate the search for a causative gene. PMID- 12761659 TI - Copper(I)-phenolate complexes as models of the reduced active site of galactose oxidase: synthesis, characterization, and O2 reactivity. AB - The Cu(I)-phenolate complexes (1)LCu and (2)LCu and the Cu(I)-phenol complex [H(2)LCu(CNC(6)H(3)Me(2))]BArF(4) were prepared and structurally characterized by X-ray crystallography, where (1)L(-) and (2)L(-) are ligands comprised of a 2,4 di- tert-butylphenolate linked to 1-isopropyl-1,5-diazacyclooctane or 1,4 diisopropyl-1,4,7-triazacyclononane, respectively. The reduced galactose oxidase (GAO) structural models (1)LCu and (2)LCu were found to be highly reactive with O(2), and through combined stopped-flow kinetic and EPR, UV-vis, and resonance Raman spectroscopic studies of the oxygenation of (2)LCu at low temperature, new intermediates relevant to those postulated for the active site oxidation step of the GAO catalytic cycle were identified. The oxygenation was shown by kinetics experiments to proceed via initial binding of O(2) to yield a green, unusually thermodynamically stable 1:1 adduct, (2)LCu(O(2)). Symmetric (eta(2)) binding of a superoxo ligand was indicated by oxygen-isotope-sensitive features in resonance Raman spectra obtained in batch experiments; peaks at nu((16)O(2))=1120 cm(-1), nu((18)O(16)O)=1093 cm(-1), and nu((18)O(2))=1058 cm(-1) were assigned as O-O stretching vibrations. These data represent the first experimental evidence for such superoxide coordination in complexes of tetradentate tripodal ligands and provide new precedent for how O(2) may bind at the reduced GAO active site. The 1:1 Cu/O(2) adduct subsequently evolves into a metastable purple species that is only observable under conditions of substoichiometric O(2). The kinetics of formation of this transient species are second order overall (rate= k'(2)[(2)LCu(O(2))][(2)LCu]). It exhibits an absorption band with lambda(max)=565 nm (epsilon=17900 M(-1) cm(-1)) and multiple oxygen-isotope-sensitive nu(Cu-O) and nu(O-O) features in the respective regions 500-550 cm(-1) and 700-850 cm(-1) in Raman spectra, with excitation-wavelength-dependent intensities that correlate with the 565 nm absorption feature. On the basis of the combined data available, the presence of multiple isomeric peroxodicopper species in the transient purple solution is postulated. PMID- 12761660 TI - Structural analysis of zinc-substituted cytochrome c. AB - Zinc-substituted cytochrome c has been widely used in studies of protein-protein interactions and photo-induced electron transfer reactions between proteins. However, the coordination geometry of zinc in zinc-substituted cyt c has not yet been determined; two different opinions about the coordination have been reached. Here the solution structures of zinc-substituted cytochrome c that might be five coordinated and six-coordinated have been refined separately by using (1)H NMR spectroscopy, and the zinc coordination geometry was determined just by NOE distance constraints. Structural analysis of the energy-minimized average solution structures of both the pentacoordinated and hexacoordinated geometries indicate that that zinc in zinc-substituted cyt c should be bound to both His18 and Met80, which means that the zinc is six-coordinated. RMSD values of the family of 25 six-coordinated structures from the average structure are 0.66+/ 0.13 A and 1.09+/-0.16 A for the backbone and all heavy atoms, respectively. A statistical analysis of the structure indicates its satisfactory quality. Comparison of the solution structure of the six-coordinated energy-minimized average structure of zinc-substituted cytochrome c with the solution structure of reduced cytochrome c reveals that for the overall folding the secondary structure elements are very close. The availability of the structure provides for a better understanding of the protein-protein complex and for electron transfer processes between Zn cyt c and other metalloproteins. PMID- 12761661 TI - Design of copper DNA intercalators with leishmanicidal activity. AB - The complexes [Cu(dppz)(NO(3))]NO(3) (1), [Cu(dppz)(2)(NO(3))]NO(3) (2), [Cu(dpq)(NO(3))]NO(3) (3), and [Cu(dpq)(2)(NO(3))]NO(3) (4) were synthesized and characterized by elemental analysis, FAB-mass spectrometry, EPR, UV, and IR spectroscopies, and molar conductivity. DNA interaction studies showed that intercalation is an important way of interacting with DNA for these complexes. The biological activity of these copper complexes was evaluated on Leishmania braziliensis promastigotes, and the results showed leishmanicidal activity. Preliminary ultrastructural studies with the most active complex (2) at 1 h revealed parasite swelling and binucleated cells. This finding suggests that the leishmanicidal activity of the copper complexes could be associated with their interaction with the parasitic DNA. PMID- 12761662 TI - A density functional investigation of the extradiol cleavage mechanism in non heme iron catechol dioxygenases. AB - The mechanism for extradiol cleavage in non-heme iron catechol dioxygenase was modelled theoretically via density functional theory. Based on the Fe(II) His,His,Glu motif observed in enzymes, an active site model complex, [Fe(acetate)(imidazole)(2)(catecholate)(O(2))](-), was optimized for states with six, four and two unpaired electrons (U6, U4 and U2, respectively). The transfer of the terminal atom of the coordinated dioxygen leading to "ferryl" Fe=O intermediates spontaneously generates an extradiol epoxide. The computed barriers range from 19 kcal mol(-1) on the U6 surface to approximately 25 kcal mol(-1) on the U4 surface, with overall reaction energies of +11.6, 6.3 and 7.1 kcal mol(-1) for U6, U4 and U2, respectively. The calculations for a protonated process reveal the terminal oxygen of O(2) to be the thermodynamically favoured site but subsequent oxygen transfer to the catechol has a barrier of approximately 30-40 kcal mol(-1), depending on the spin state. Instead, protonating the acetate group gives a slightly higher energy species but a subsequent barrier on the U4 surface of only 7 kcal mol(-1) relative to the hydroperoxide complex. The overall exoergicity increases to 13 kcal mol(-1). The favoured proton-assisted pathway does not involve significant radical character and has features reminiscent of a Criegee rearrangement which involves the participation of the aromatic ring pi orbitals in the formation of the new carbon-oxygen bond. The subsequent collapse of the epoxide, attack by the coordinated hydroxide and final product formation proceeds with an overall exoergicity of approximately 75 kcal mol(-1) on the U4 surface. PMID- 12761663 TI - Multifrequency cw-EPR investigation of the catalytic molybdenum cofactor of polysulfide reductase from Wolinella succinogenes. AB - Electron paramagnetic resonance (EPR) spectra of the molybdenum centre in polysulfide reductase (Psr) from Wolinella succinogenes with unusually high G tensor values have been observed for the first time. Three different Mo(V) states have been generated (by the addition of the substrate polysulfide and different redox agents) and analysed by their G- and hyperfine tensors using multifrequency (S-, X- and Q-band) cw-EPR spectroscopy. The unusually high G-tensor values are attributed to a large number of sulfur ligands. Four sulfur ligands are assumed to arise from two pterin cofactors; one additional sulfur ligand was identified from mutagenesis studies to be a cysteine residue of the protein backbone. One further sulfur ligand is proposed for two of the Mo(V) states, based on the experimentally observed shift of the g(av) value. This sixth sulfur ligand is postulated to belong to the polysulfide substrate consumed within the catalytic reaction cycle of the enzyme. The influence of the co-protein sulfur transferase on the Mo(V) G-tensor supports this assignment. PMID- 12761664 TI - Interactions of the pyridine-2-carboxaldehyde isonicotinoyl hydrazone class of chelators with iron and DNA: implications for toxicity in the treatment of iron overload disease. AB - Iron chelation therapy for the management of iron-overload disease is dominated by desferrioxamine (DFO). However, treatment using DFO is very arduous. Recently, novel Fe chelators of the pyridine-2-carboxaldehyde isonicotinoyl hydrazone (PCIH) class have shown high chelation efficacy and the potential to replace DFO. A critical consideration in the design of alternatives to DFO is that the chelator forms a redox-inert Fe complex. In the present study, the participation of Fe complexes in redox reactions has been investigated. Ascorbate oxidation in the presence of Fe(III) or benzoate hydroxylation in the presence of Fe(II) was not enhanced by the PCIH analogues. However, redox-induced DNA strand breaks were observed with these ligands under highly oxidizing conditions in the presence of Fe(II) and hydrogen peroxide. Experiments then examined the interactions of the PCIH analogues with DNA, and this was found to be weak. Considering this, we suggest that under extreme conditions seen in the DNA-strand break assay, weak DNA-binding may potentiate the redox activity of the PCIH analogues. However, importantly, in contrast to naked plasmid DNA, DNA damage by these chelators using intact human cells was not significant. Collectively, our results support the potential of the PCIH analogues for the treatment of Fe overload. PMID- 12761665 TI - Raman spectra and normal coordinate analyses of low-frequency vibrations of oxo bridged manganese complexes. AB - The active sites of certain metalloenzymes involved in oxygen metabolism, such as manganese catalase and the oxygen-evolving complex of photosystem II, contain micro -oxo-bridged Mn clusters with ligands that include H(2)O and micro (1,3) carboxylato bridges provided by protein side chains. In order to understand better the vibrational spectra of such clusters, the low-frequency resonance Raman spectra of a series of structurally characterized Mn-oxo model complexes were examined. The series includes complexes of the type [Mn(2)(O)(OAc)(2)(bpy)(2)(L)(2)] and [Mn(2)(O)(2)(OAc)(bpy)(2)(L)(2)], where bpy=2,2'-bipyridine, OAc=acetate and L=H(2)O or Cl(-). Complexes containing the isotopomers OAc- d(3) and D(2)O, as well as those containing both isotopomers, were also examined. Normal coordinate analyses (NCA) were performed on the various complexes using theGF matrix method. Selected vibrational modes in the 200-600 cm(-1) region were assigned based on the spectra and NCA, which identify vibrational modes arising from the metal-ligand bonds. These results will be useful in interpreting the vibrational spectra obtained from metalloproteins containing Mn-oxo complexes in their active sites. PMID- 12761666 TI - Metal binding to Bacillus subtilis ferrochelatase and interaction between metal sites. AB - Ferrochelatase, the terminal enzyme in heme biosynthesis, catalyses metal insertion into protoporphyrin IX. The location of the metal binding site with respect to the bound porphyrin substrate and the mode of metal binding are of central importance for understanding the mechanism of porphyrin metallation. In this work we demonstrate that Zn(2+), which is commonly used as substrate in assays of the ferrochelatase reaction, and Cd(2+), an inhibitor of the enzyme, bind to the invariant amino acids His183 and Glu264 and water molecules, all located within the porphyrin binding cleft. On the other hand, Mg(2+), which has been shown to bind close to the surface at 7 A from His183, was largely absent from its site. Activity measurements demonstrate that Mg(2+) has a stimulatory effect on the enzyme, lowering K(M) for Zn(2+) from 55 to 24 micro M. Changing one of the Mg(2+) binding residues, Glu272, to serine abolishes the effect of Mg(2+). It is proposed that prior to metal insertion the metal may form a sitting atop (SAT) complex with the invariant His-Glu couple and the porphyrin. Metal binding to the Mg(2+) site may stimulate metal release from the protein ligands and its insertion into the porphyrin. PMID- 12761667 TI - Synthesis of a new vanadyl(IV) complex with trehalose (TreVO): insulin-mimetic activities in osteoblast-like cells in culture. AB - Vanadium compounds show interesting biological and pharmacological properties. Some of them display insulin-mimetic effects and others produce anti-tumor actions. The bioactivity of vanadium is present in inorganic species like the vanadyl(IV) cation or vanadate(V) anion. Nevertheless, the development of new vanadium derivatives with organic ligands which improve the beneficial actions and decrease the toxic effects is of great interest. On the other hand, the mechanisms involved in vanadium bioactivity are still poorly understood. A new vanadium complex of the vanadyl(IV) cation with the disaccharide trehalose (TreVO), Na(6)[VO(Tre)(2)].4H(2)O, here reported, shows interesting insulin mimetic properties in two osteoblast cell lines, a normal one (MC3T3E1) and a tumoral one (UMR106). The complex affected the proliferation of both cell lines in a different manner. On tumoral cells, TreVO caused a weak stimulation of growth at 5 microM but it inhibited cell proliferation in a dose-response manner between 50 and 100 microM. TreVO significantly inhibited UMR106 differentiation (15-25% of basal) in the range 5-100 microM. On normal osteoblasts, TreVO behaved as a mitogen at 5-25 microM. Different inhibitors of the MAPK pathway blocked this effect. At higher concentrations (75-100 microM), the complex was a weak inhibitor of the MC3T3E1 proliferation. Besides, TreVO enhanced glucose consumption by a mechanism independent of the PI3-kinase activation. In both cell lines, TreVO stimulated the ERK phosphorylation in a dose- and time-dependent manner. Different inhibitors (PD98059, wortmannin, vitamins C and E) partially decreased this effect, which was totally inhibited by their combination. These results suggest that TreVO could be a potential candidate for therapeutic treatments. PMID- 12761668 TI - Docking and electron transfer studies between rubredoxin and rubredoxin:oxygen oxidoreductase. AB - The interaction and electron transfer (ET) between rubredoxin (Rd) and rubredoxin:oxygen oxidoreductase (ROO) from Desulfovibrio gigas is studied by molecular modelling techniques. Experimental kinetic assays using recombinant proteins show that the Rd reoxidation by ROO displays a bell-shaped dependence on ionic strength, suggesting a non-trivial electrostatic dependence of the interaction between these two proteins. Rigid docking studies reveal a prevalence for Rd to interact, in a very specific way, with the surface of the ROO dimer near its FMN cofactors. The optimization of the lowest energy complexes, using molecular dynamics simulation, shows a very tight interaction between the surface of the two proteins, with a high probability for Rd residues (but not the iron centre directly) to be in direct contact with the FMN cofactors of ROO. Both electrostatics and van der Waals interactions contribute to the final energy of the complex. In these complexes, the major contributions for complex formation are polar interactions between acidic residues of Rd and basic residues of ROO, plus substantial non-polar interactions between different groups. Important residues for this process are identified. ET estimates (using the Pathways model), in the optimized lowest energy complexes, suggest that these configurations are efficient for transferring electrons. The experimental bell shaped dependence of kinetics on ionic strength is analysed in view of the molecular modelling results, and hypotheses for the molecular basis of this phenomenon are discussed. PMID- 12761669 TI - Do autologous growth factors enhance transforaminal lumbar interbody fusion? AB - Pseudarthrosis remains a significant problem in spinal fusion. The objective of our study was to investigate the effects of autologous growth factors (AGF) in instrumented transforaminal lumbar interbody spinal fusion (TLIF). A prospective review was carried out of 23 patients who underwent TLIF with application of AGF, with a minimum 2-year follow-up. Comparison with our historical cohort (without AGF application) was performed. Mean age at surgery was 44.3 years in the AGF treatment group. Twelve had a positive smoking history. Fourteen had undergone previous spinal surgeries. Thirteen received one-level fusions and ten received two-level fusions. The radiographic results showed a fusion rate of 100% in one level fusions and 90% in two-level fusions. There was no significant difference in pseudarthrosis rates between the AGF treatment group and historical cohort. Excluding the cases with pseudarthrosis, there was faster bony healing in patients who had been treated with AGF application. This study indicates that although AGF may demonstrate faster fusions, it does not result in an overall increase in spinal fusion rates. Further studies are needed before AGF can routinely be used as an adjunct in spinal fusion. PMID- 12761671 TI - Experimental evidence for physical cues involved in oviposition site selection of lotic hydrobiosid caddis flies. AB - Cues involved in determining the distribution of invertebrate propagules within a stream landscape contribute greatly to our knowledge of the supply and arrangement of new recruits and thus an improved understanding of factors that might ultimately affect population parameters. Previous observations indicated that both current velocity and rock size were important determinants of the egg mass distribution of certain hydrobiosid caddis flies that lay their eggs in single masses beneath emergent rocks. These observations were tested experimentally in a temperate, upland Australian stream. Manipulations of current speed confirmed that females of Ulmerochorema sp. and the Taschorema complex deposited more eggs on rocks in elevated current speeds (>0.40 ms(-1)) whereas Apsilochorema sp. deposited more eggs on rocks in slow currents (<0.30 ms(-1)). This latter result did not coincide with previous observational data. The anomaly between observational and experimental data, however, was reconciled by the outcome of a further experiment that tested the influence of the emergent or 'landing pad' size of rocks as the abundance of Apsilochorema egg masses increased with landing pad size independent of the prevailing flow conditions. Landing pad size did not influence the abundance of egg masses of Taschorema or Ulmerochorema. Patterns of female visits to rocks indicated that taxa might distinguish between favoured egg-laying sites prior to landing on rocks. Large aggregations of adult male and female Ulmerochorema collected from rocks favoured for oviposition provide indirect evidence for mating induced swarming behaviour associated with oviposition site selection. This study provides a framework for more sophisticated questions relating to the influence of oviposition site selection on structuring populations of lotic macroinvertebrates. PMID- 12761670 TI - Genetic and metabolic determinants of increased plasma plasminogen activator inhibitor-1 activity in children with renal transplants. AB - Recent studies have shown that activity of plasminogen activator inhibitor-1 (PAI 1), a prothrombotic protein, may be increased in transplanted patients. The aim of the present investigation was to determine PAI-1 activity in pediatric recipients of renal transplants and to establish the relative contribution of both genetic and metabolic factors. In 29 children and adolescents with stable renal transplants, we related plasma PAI-1 activity to an indicator of inflammatory status [plasma concentration of C-reactive protein (CRP)] and to elements of the insulin resistance syndrome [body mass index (BMI), fasting insulinemia, HOMA index and plasma triglyceride, HDL-cholesterol, apolipoproteins A-1 and B concentrations]. Polymorphisms of PAI-1, apolipoprotein E (apoE) and angiotensin-converting enzyme (ACE) genes were also investigated. In all patients the study was repeated 1 year later. PAI-1 activity remained constantly elevated (23.4+/-22.8 and 18.6+/-7.8 U/ml in the first and second study, respectively, P=NS). Plasma PAI-1 activity correlated positively with CRP ( P=0.001), BMI z score ( P=0.02), fasting insulinemia ( P=0.009), and HOMA index ( P=0.006). No significant correlations were found in this population between plasma PAI-1 activity and age, gender, time elapsed after transplantation and plasma homocysteine, total cholesterol, LDL-cholesterol, HDL-cholesterol, apolipoprotein B, and apolipoprotein A-1. Plasma PAI-1 activity was not related to the cumulative dose of prednisone, cyclosporin A, or tacrolimus. Plasma PAI-1 activity was significantly higher in 5 children with apoE3/apoE4 genotype. No apparent influences of the PAI-1 4G/4G and ACE I/D genotypes were observed. In a multiple stepwise regression model, fasting insulinemia and apoE3/apoE4 genotype accounted for 45% of the observed plasma PAI-1 variability. We conclude that increased PAI-1 activity in children with stable renal transplants is determined both by genetic factors and by metabolic factors, the latter mainly linked to the insulin resistance syndrome. PMID- 12761672 TI - The divergent expression of periostin mRNA in the periodontal ligament during experimental tooth movement. AB - A novel 90-kDa protein named periostin, which is preferentially expressed in the periosteum and the periodontal ligament (PDL), may play a role in bone metabolism and remodeling. However, the precise role of periostin in the PDL remains unclear. Therefore, we examined the expression of periostin mRNA during experimental tooth movement. Experimental tooth movement was achieved in 7-week old male Sprague-Dawley rats. In control specimens without tooth movement, the expression of periostin mRNA was uniformly observed in the PDL surrounding the mesial and distal roots of the upper molars and was weak in the PDL of the root furcation area. The periostin mRNA-expressing cells were mainly fibroblastic cells in the PDL and osteoblastic cells on the alveolar bone surfaces. The divergent expression of periostin mRNA in the PDL began to be observed at 3 h and continued up to 96 h after tooth movement. The maximum changes, which showed stronger staining in the pressure sites than in the tension sites, were observed at 24 h. The expression of periostin mRNA in the PDL 168 h after tooth movement exhibited a similar distribution to that of the control specimens. These results suggest that periostin is one of the local contributing factors in bone and periodontal tissue remodeling following mechanical stress during experimental tooth movement. PMID- 12761673 TI - Epidemiology of herpes simplex virus types 1 and 2 in Germany: what has changed? AB - Infections with herpes simplex virus (HSV) types 1 and 2 are widespread in all human populations and result in persistent and latent infections. HSV-1 is commonly responsible for orofacial, HSV-2 more likely causes genital lesions. Herpes genitalis is one of the most important sexually transmitted diseases; furthermore, there are severe diseases associated with HSV (e.g., encephalitis). Over the last years an increase in clinical manifestations of HSV has been reported, and HSV-1 has been increasingly discussed as causative agent of herpes genitalis. We retrospectively evaluated the laboratory results of our routine diagnostic service for HSV infections, looking for changes of HSV epidemiology in recent years. Specimens from 2,678 herpes patients were obtained between 1 January 1996 and 31 March 2002. Using cell culture, the presence of HSV was investigated in swabs taken from different body sites, and clinical data on HSV localization and type were evaluated. We found 345 patients positive for HSV-1 and 212 positive for HSV-2. Clinical data were available from 72.1% of the patients with HSV-1, and 61.3% of those with HSV-2 infection. In genital herpes HSV-1 was the causative agent in 20% of men and in 25% of women. In patients suffering from orofacial herpes HSV-2 was detected in 7% of men and in 4% of women. To evaluate the frequency of neurological HSV diseases, 2,406 cerebrospinal fluid samples (CSF) from 2,121 patients suspected of meningitis or encephalitis were tested for HSV DNA by the polymerase chain reaction. Among those patients, 120 showed CSF positive for HSV DNA. Serum surveys of HSV-1 and HSV-2 infection recently established in our region were compared to similar studies performed in Germany 25 years ago. We found that seroprevalences have not changed over the last 25 years and that neurological HSV diseases are rare. However, as in the USA, a significant percentage of herpes genitalis is caused by HSV-1 in Germany. PMID- 12761674 TI - Neuronavigation combined with electrophysiological monitoring for surgery of lesions in eloquent brain areas in 42 cases: a retrospective comparison of the neurological outcome and the quality of resection with a control group with similar lesions. AB - The purpose of this study was to achieve a more radical resection of tumors in the area of the motor cortex via minimal craniotomy using a combination of neuronavigation and neurophysiological monitoring with direct electrical cortical stimulation and to compare retrospectively the clinical outcome and postoperative magnetic resonance imaging with a control group that was operated on in our service when the combination of these monitoring techniques was not available. A total of 42 patients with tumors in or near the central region underwent surgery with neuronavigation guidance and neurophysiological monitoring. The primary motor cortex was identified intraoperatively by the somatosensory evoked phase reversal method and direct cortical stimulation. The functional areas were transferred into the neuronavigation system. By stimulating the identified primary motor cortex and displaying the motor area in the operating microscope a permanent control of the motor function was possible during the whole operation. Using these techniques a more radical tumor resection - evaluated by postoperative MRI - was achieved in the study group (p = 0.04) and also a trend toward a better neurological outcome. PMID- 12761675 TI - Image-guided removal of supratentorial cavernomas in critical brain areas: application of neuronavigation and intraoperative magnetic resonance imaging. AB - In a retrospective study the postoperative results of 26 patients operated on for supratentorial cavernous hemangiomas either deep-seated or near eloquent brain areas are summarized. An exact surgical approach to these lesions is essential to prevent neurological deterioration. Three different navigation systems were used and compared according to their clinical applicability. Complete removal of the lesion was obtained in all patients of this series. In six cases (23 %) functional data from magnetoencephalography or functional magnetic resonance imaging were integrated into the navigational setup. In 14 cases (54 %) intraoperative magnetic resonance imaging was performed. The follow-up time was 3 - 26 months (mean: 10 months). In the postoperative course one patient (3.8 %) developed a hemiparesis, another one developed quadrantopia. Nineteen patients presented with preoperative seizure history, 16 of these (84 %) had no further or rare seizures after surgery. The better results in seizure control were achieved in those patients with shorter duration of seizure history before surgery. The study indicates that the application of neuronavigation allows surgery on supratentorial cavernous hemangiomas in critical brain areas with low morbidity. The intraoperative visualization of eloquent cortex areas by integration of functional data allows a fast identification and exemption of eloquent brain areas, preventing neurological deterioration. Furthermore, the intraoperative MR resection control ensures a complete resection and illustrates the minimal invasive approach. PMID- 12761676 TI - Image-guided transsphenoidal surgery for pituitary lesions using Mehrkoordinaten Manipulator (MKM) navigation system. AB - Twenty-five patients with pituitary lesions were operated on by image-guided transsphenoidal surgery (TSS) using the Mehrkoordinaten Manipulator (MKM) navigation system. The cases included 21 cases of pituitary adenomas, 2 cases of craniopharyngioma and 2 cases of Rathke's cleft cyst. All operations were performed through the sublabial approach under an operative microscope. In some cases, an endoscope was used for the observation of the residual tumor and surrounding structures. The tumors and surrounding important structures such as the internal carotid arteries, the basilar artery, and the optic nerves were precisely localized, and mechanical error was less than 2 mm in almost all cases. In 3 early cases of pituitary adenoma, the patient's head was moved slightly during the insertion of the nasal speculum; in these cases, the resulting error was more than 2 mm. In evaluating the procedures, we determined that the most useful benefit of the MKM system compared with other systems is that the navigation information is not only displayed on the monitor, but also presented in the operative field under the microscope. Therefore, the surgeon can obtain the navigation information without removing his eyes from the operative field under the microscope. The most important drawback to the system is its bulky size. PMID- 12761677 TI - Frameless stereotactic radiosurgery with mobile CT, mask immobilization and micro multileaf collimators. AB - We present frameless stereotactic radiosurgery using mobile CT, thermal plastic mask fixation, a vacuum-form body immobilizer and micro-multileaf collimators. A linear accelerator and a self-moving helical CT scanner gantry were installed in the same room. The isocenter of irradiation can be also aligned with the center of the CT gantry by rotating the couch. A thermal plastic mask and vacuum-form body immobilizer was used for registration and immobilization. The advantages of this system are as follows: 1) Accurate and painless frameless fractionated irradiation can be performed smoothly, as the patient's head is fixed without exchanging the couch from CT scanning to irradiation system. 2) This mask system can be applied to children, infants or adults with a previous craniotomy bone flap that must have a fixation pin placed into it. This system can be also used for fractionated radiotherapy without painful skull pin-fixed frame. 3) 1 mm micro-multileaf collimators enable irregular contour irradiation. 4) Image fusion (among CT, MRI, angiography, and PET) and 3D images can be used for irradiation planning. 5) This system can be used on any part of the body. 6) This system can be installed in any irradiation room without any extension or new construction. PMID- 12761678 TI - Radiobiological effects of photon radiosurgery on choroid plexus cells in vitro. AB - The standard surgical treatment of hydrocephalus by cerebrospinal fluid (CSF) shunt is accompanied by numerous complications. The search for alternative treatment methods includes resection, coagulation and irradiation of part of the plexus choroideus. The reduction of CSF secretion after choroid plexus (CP) irradiation has been investigated only on the experimental level. The new Photon Radiosurgery System (PRS) now also provides clinically the opportunity to induce selective radionecrosis on the CP with high efficiency and safety. In order to achieve a basic understanding of the reaction of CP cells after PRS irradiation, we investigated the cell death after different irradiation doses using TB dye exclusion and MTT assay on sheep choroid plexus (SCP) cells. We observed a dose dependent decrease in cell survival with increasing doses of irradiation (9, 18, 27 and 36 Gy). Lower irradiation doses (9, 18 Gy) induced an initial decrease of cell survival. Cells were able to recover from day 6 on and achieved a similar cell viability compared to non-irradiated cells on day 12. In contrast, higher doses (27 and 36 Gy) of irradiation induced a constant decrease of the cell survival over 12 days. These results clearly demonstrate that PRS irradiation is able to induce radionecrosis of CP cells which are responsible for the secretion of CSF. Interstitial photon radiosurgery can provide the opportunity to deliver the irradiation dose locally to CP with minimal exposure of surrounding tissue. Our basic data support further studies investigating this concept in animal models and clinically. PMID- 12761679 TI - Stereotactic biopsy in the era of advanced neuroimaging. Does the minimal therapeutic gain justify its current wide use? AB - OBJECTIVE: This study reviewed the contribution of stereotactic brain biopsy in the management and final outcome of a series of patients with presumed inoperable lesions. PATIENTS AND METHODS: Sixty-nine consecutive patients underwent a CT guided lesion biopsy (n = 67) or abscess/cyst aspiration (n = 2) using the Cosman Roberts-Wells (CRW) frame. RESULTS: A definitive specific diagnosis was made in 53 of 67 patients (79 %). The remaining procedures did not provide a diagnosis because of failure to obtain appropriate specimen (11 patients), findings consistent with non-specific inflammation (2 patients) or uneventful surgical complication requiring termination of the procedure (1 patient). A total of 55 patients (80 %) died due to the malignant nature of the lesion, most within six months after the biopsy. The preoperative imaging diagnosis was consistent with the histological diagnosis in 60 patients (87 % accuracy). The perioperative morbidity and mortality were nil and most of the patients were discharged within twenty-four hours. CONCLUSIONS: The stereotactic biopsy did not alter either the therapeutic management or the mortality due to the natural course of the lesion. These findings indicate that the current principle of mandatory histological diagnosis in virtually all non-resectable brain lesions should be re-evaluated taking into account parameters such as: age, medical/neurological status, neuroimaging characteristics, patients' best interest and health care financial shortages. PMID- 12761680 TI - Thecaloscopy part III: first clinical application. AB - PURPOSE: We have already dealt with the technical and anatomical basics for thecaloscopy. The first results of clinical application are presented and the indications for its diagnostic and therapeutic use are described. The aetiology of aseptic arachnitis and the term "Wide spinal canal syndrome" are discussed. PATIENTS AND METHODS: Twelve patients who fulfilled the criteria for entry to the study were selected and the criteria were chosen according to German medical and legal rules. All patients underwent an endoscopic inspection and, if necessary, therapeutic manipulation under general anaesthesia. RESULTS: In all cases the procedure was safely and successfully performed. The pathomorphology of the arachnoid was detected and endoscopically treated in almost all cases. Arachnoid cysts were successfully fenestrated and an intraspinal meningocoele treated with endoscopic assistance. CONCLUSION: Thecaloscopy is a safe procedure if skilfully performed. It provides an opening for a wide range of new diagnostic and therapeutic options. PMID- 12761681 TI - The extradural subtemporal keyhole approach to the sphenocavernous region: anatomic considerations. AB - Fourteen cadaver specimens (28 sides) and twelve dry human skulls (24 sides) were used to study the anatomic relationships between bony, neurovascular and foraminal landmarks in the floor of the middle fossa in preparation for performing the extradural subtemporal keyhole approach to the sphenocavernous region. The interforaminal distance was largest between the foramina rotundum (FR) and ovale (FO) and was smallest between the FO and foramen spinosum (FS). The largest angle between exit foramen was the FR to FO. The greater superficial petrosal nerve (GSPN) was always found to overlie and run parallel to the petrous internal carotid artery, however, its location over the artery and its separation from it by bone was variable. With a subtemporal "keyhole" placed above the posterior zygomatic root (PZR), a 0 degrees endoscope allowed easy visualization of the middle meningeal artery (MMA) and the mandibular nerve (V 3 ) however, a 30 degrees endoscope was more useful for visualizing the maxillary nerve (V 2 ) and the ophthalmic nerve (V 2 ). With a sphenoidotomy performed between V 1 and V 2, the 30 degrees endoscope was found to be the most useful for visualizing the carotid siphon and the contralateral wall of the sphenoid sinus, while the 70 degrees endoscope was the most useful for visualizing of the floor of the sella and the walls of the sphenoid sinus. Two venous concerns with respect to performing endoscopic approaches to the region were identified: a fibrous layer overlies a heavy venous plexus that encircles the petrous carotid artery, and the foramen Vesalius, which transmits a large emissary vein draining the cavernous sinus, was identified medial to the FO in 30 % of our dissected sides. PMID- 12761683 TI - Spontaneous disappearance of two asymptomatic arachnoid cysts in two different locations. AB - We report two children with asymptomatic arachnoid cysts which resolved spontaneously without any surgical intervention and history of major head and body trauma. The first child was a 10-year-old boy with an arachnoid cyst in the right sylvian fissure. The second child was a 1-year-old girl with a right cerebral convexity arachnoid cyst. Both of them were asymptomatic. Arachnoid cysts spontaneously disappeared within 2 years following initial diagnosing. There was no major head and body trauma except usual home, school and sports activity. We speculated that the cysts ruptured into cerebrospinal fluid circulation by the mechanical effects of some forced activities to the brain tissue and cyst, such as excessive breathing, coughing and sport activities. These factors may change the balance between intracystic and pericystic pressure and facilitate the rupturing of the cyst into subdural, subarachnoid and intraventricular spaces. These cases demonstrate that neurosurgical intervention of asymptomatic arachnoid cysts is not absolutely indicated in the paediatric age group. Close follow up with computerized tomography (CT) and magnetic resonance imaging (MRI) is a treatment option in the patient with arachnoid cysts located in the middle cranial fossa and cerebral convexity. PMID- 12761682 TI - Influence of TachoComb in comparison to local hemostyptic agents on epidural fibrosis in a rat laminectomy model. AB - This animal experimental study was designed to examine the effects of TachoComb, a fixed combination of collagen with tissue adhesive, as an interposition membrane on the development of spinal epidural fibrosis in comparison to other hemostyptic materials. In 10 Wistar rats, four laminectomies were performed at lumbar and sacral vertebrae. Alternately, a piece of TachoComb, Spongostan, or Tabotamp was placed into each laminectomy site. One laminectomy site served as an empty control (n = 10). 8 weeks later, the animals were sacrificed, and the spinal column including surrounding muscle tissue was removed en bloc from each rat and fixed in formaldehyde. After decalcification and staining the specimens were graded by a neuropathologist in a blindfold test for severity of epidural fibrosis as "light-moderate" or "marked". Epidural scarring of variable density was seen in all laminectomy sites. Light epidural fibrosis, without any adhesion to dura, as only noted in cases after application of TachoComb (n = 4/10) and Spongostan (n = 1/10). All other slices showed marked epidural fibrosis with dura adherence regardless of the implanted material. Statistical analysis revealed significantly lower epidural fibrosis after application of TachoComb compared to all other groups (p < 0.05). In this series, TachoComb is more effective in reducing the epidural fibrosis than Spongostan, and Tabotamp. However, complete prevention of scar tissue formation was not achieved. PMID- 12761684 TI - Unusual intraventricular herniation of the suprasellar arachnoid cyst and its successful endoscopic management. AB - A child with a benign intrinsic tectal tumor and obstructive hydrocephalus developed a huge intraventricular cystic lesion following insertion of a shunt, which was complicated by a subdural-subarachnoid bleeding. The lesion was identified to be an entrapped chiasmatic cistern, which disrupted the septum pellucidum in front of the third ventricle and filled gradually the anterior horn of the lateral ventricle. The condition was successfully treated endoscopically by fenestration of the cyst and a third ventriculostomy. Probable mechanisms of such unusual cyst formation and general management strategies in patients with obstructive hydrocephalus are discussed. PMID- 12761685 TI - Updating of neuronavigation based on images intraoperatively acquired with a mobile computerized tomographic scanner: technical note. AB - Image-guided surgery based on preoperatively obtained image data is susceptible to inaccuracy resulting from intraoperative brain shift and distortion. We report a technique of updating neuronavigational data with the aid of a mobile computerized tomographic (CT) scanner. A mobile CT which is readily available in an ordinary operating room was used to acquire intraoperative images. A total of 6 - 7 titanium screws placed on the skull were used as new reference points for updating navigation. Intraoperative CT scanning was performed with a 2 mm slice thickness. After the obtained image data were transferred as Dicom files to the computer workstation of the navigation system through an Ethernet connection, navigational data were updated to registering the new reference points. Under the guidance of the updated navigation, residual lesions were explored, and further resected. Our preliminary experience in 8 patients indicates that interactive image-guidance can stably be updated based on images intraoperatively acquired with a mobile CT scanner. Comparing to intraoperative magnetic resonance imaging, this technique can simply be done in an ordinary operating room without requiring special surgical instruments, thus making it possible to update interactive image guidance on demand during an operation. PMID- 12761686 TI - Holmium:YAG laser-induced liquid jet dissector: a novel prototype device for dissecting organs without impairing vessels. AB - BACKGROUND AND OBJECTIVE: Neurosurgery has long required a method for dissecting brain tissue without damaging principal vessels and adjacent tissue, so as to prevent neurological complications after operation. In this study we fabricated such a prototype device and used it in an attempt to resect an animal liver, which, like the brain, contains many vessels. MATERIALS AND METHODS: The prototype device consisted of a jet nozzle and a suction tube. Pulsed liquid jets at 3 Hz were ejected from the nozzle by a pulsed holmium:YAG (Ho:YAG) laser at an irradiation energy of 230 mJ/pulse. The profile of the liquid jet was observed with a high-speed camera. With this device, liver dissections of anesthetized rabbits were attempted while measuring the local temperature of the target. A histological study of the incised parts was also performed. RESULTS: The liquid jet was emitted straight from the nozzle at an initial velocity of 38 m/sec. The liver parenchyma was cut with the device while preserving the tiny vessels and keeping the operative field clear. The local temperature rose to no more than 314 K (below the heat damage threshold of brain tissue). In the histological findings, there were no signs of hepatic degeneration or necrosis around the dissected margin. CONCLUSIONS: The Ho:YAG laser-induced liquid jet dissector can be applied to neurosurgical operations after incorporating some minor improvements. PMID- 12761687 TI - [Care of persons with diabetes in Germany--where do we stand today?]. PMID- 12761688 TI - [Improvement of care for diabetics using the care model from Saxony]. AB - BACKGROUND AND OBJECTIVE: By promoting the networking of all those involved in caring for diabetics in Saxony, through agreement between those who provide help to them and the organizations which pay the costs, the intention is to improve the overall quality of care of diabetics. It was the aim of this study to evaluate the effectiveness of this integrated health model. PATIENTS AND METHODS: As part of the 3rd Diabetes Agreement in Saxony, a total of 275,804 diabetics were registered, treated and their management costed in the first quarter of 2000 and the fourth quarter of 2001 (56.3% females, 43.7% males; median age 68,7 years). They were patients of 2800 general practitioners and 88 specialist practices. RESULTS: Nearly 80% of all diabetics were included. Taking the level of HbA1c as the criterion of quality achieved, it had decreased from 7,1 +/- 1,3 % in the first 3 months of 2000 to 6,8 +/- 1,3 % in the last 3 months of 2001, and regional differences had been reduced. There was an obvious correlation between early referral to specialist practices and good treatment results, as measured by HbA1c and blood pressure levels. While in 1996 patients were referred when the HbA1c level was 8.8% (median 8.5%), referrals in the last quarter of 2001 were made when the mean was 8,0% (median 7.7%). After two years the risk of inadequate treatment (HbA1c > 7.5% and blood pressure > 140/90 mmHg) had been clearly reduced in about half the cases. CONCLUSIONS: Diabetes agreements, as promulgated in Saxony, have provided effective disease management programs (DMP) for efficacious and efficient integrated diabetic care, so that with continuing effectiveness and further development the St. Vincent targets can be reached. Successful regional diabetes agreements must therefore be maintained within the new, politically centralized, DMPs. PMID- 12761689 TI - [Disease management program (DMP) diabetes mellitus: simulation of therapeutic results of different guidelines. A new diabetes mellitus model (DMM)]. AB - BACKGROUND AND OBJECTIVE: In conjunction with the introduction of disease management programmes in Germany there is an ongoing scientific debate on the desirable goals for HbA1c in the management of patients with type 2 diabetes mellitus. PATIENTS AND METHODS: In this study, a novel computer-based simulation model (diabetes mellitus model = DMM) was used to estimate the consequences of different levels of metabolic control as assessed by HbA1c for the development of short- and long-term complications of this disease. RESULTS: At a mean difference of 1 % the rate of severe hypoglycaemic events over 10 years was by 32-84 % higher in those with a lower HbA1c. In contrast, the incidence of microvascular complications (proliferative retinopathy, end-stage kidney disease, clinical neuropathy) was by 20-33 % lower in the group with the lower as compared to the higher HbA1c level according to the scenario applied. The rates of myocardial infarction and stroke were reduced by 15-20 % under these conditions. CONCLUSIONS: This model calculation suggests that a more strict metabolic control in patients with type 2 diabetes mellitus results in a greater reduction of microvascular complications than of macrovascular complications, but is associated with a higher rate of hypoglycaemic episodes. The diabetes mellitus model is a non-expensive alternative to simulate clinically relevant questions on the management of type 2 diabetes and to provide rapid and realistic answers. PMID- 12761690 TI - [Pilot study of controlled retrospective analysis (retrolective study design) of care for type 2 diabetics since 1990]. AB - BACKGROUND AND OBJECTIVE: There are few data on the quality of care received by patients with type 2 diabetes under routine conditions and only a few long-term studies that investigated the influence of treatment strategies on the development of clinical endpoints. It was the aim of this study, using retrospectively obtained data, to determine whether it is possible to document over a 10 year period the treatment design and clinical end-points in type 2 diabetics under the care of general practitioners. PATIENTS AND METHODS: General practitioners were randomly selected and informed by independent collaborating monitors about the aim of the study and data documentation. At a second visit by a monitor the completed data forms were audited and entered into a database. Course of treatment was documented by 21 general practitioners, two of them diabetes specialists, for all patients in whom, between 1.1.1990 and 31.12.1993, type 2 diabetes had been diagnosed for the first time (n=455; 251 females, 204 males, mean age 61,5 years). Continuous complete documentation of the course of treatment in the given practice was obtained in 85% of patients over a mean observation period of 8,7 years. 45 myocardial infarcts in 40 patients (9%), 42 strokes in 37 patients (8%), blindness in seven eyes of five patients (1,5%) and 19 amputations (4,2%) were recorded; 72 patients died (15,8%). CONCLUSION: The data of this pilot study indicate that detailed analysis of treatment quality can be obtained in a sizeable cohort, using of a well controlled retrospective (retrolective) study design. PMID- 12761691 TI - [Quality of prescription drug use in patients with type 2 diabetes mellitus in Germany]. PMID- 12761692 TI - [Routine screening for microalbuminemia in type 2 diabetes mellitus--pro]. PMID- 12761693 TI - [Routine screening for microalbuminemia in type 2 diabetes mellitus--con]. PMID- 12761694 TI - [Can ascorbic acid and acetylsalicylic acid distort the results of blood sugar test strips?]. PMID- 12761695 TI - [Can insulin humalog be given during pregnancy?]. PMID- 12761697 TI - Outcome in pediatric tracheotomy. AB - OBJECTIVE: To investigate the outcome and related factors in pediatric tracheotomy. DESIGN: Retrospective chart review. SETTING: Tertiary pediatric academic hospital setting. PATIENTS: The study included 181 children below the age of 18 years who underwent 185 tracheotomies between 1991 and 1995. MAIN OUTCOMES AND MEASURES: Presenting symptoms and signs, indications, duration of follow-up, therapeutic and interval procedures, early and late complications, mortality, time to and success in decannulation. RESULTS: There were 108 (59.7%) male patients and 73 (40.3%) female patients. The average age of the children at the time of tracheotomy was 3.8 +/- 5.3 years. The majority of the children were less than 1 year of age (n = 99, 54.7%). Airway obstruction was the leading indication for tracheotomy (59.6%), followed by ventilatory support (30.4%) and pulmonary toilet (9.9%). The average duration of follow-up was 931 +/- 790 days. There were no perioperative complications. Early postoperative complications were seen in 28 (15.5%) children including 12 (6.8%) major complications and 22 (12.2%) minor complications. Late complications were seen in 115 (63.5%) children, including 8 (4.4%) major complications and 107 (59.1%) minor complications. Overall mortality rate was 13.3%, but only 1 tracheotomy-related death was caused by tube displacement. Therapeutic procedures were performed in 43% of the children, including laryngotracheal reconstruction (13%), laser excision of the lesion (5%), and supraglottoplasty (3.9%). Decannulation was accomplished in 116 (64.1%) of the children with an average of 365 +/- 388 days with tracheotomy. CONCLUSION: Tracheotomy is relatively safe in the pediatric population. Decannulation may be possible relatively quickly with resolution of the underlying problem. PMID- 12761696 TI - Minimum description length block finder, a method to identify haplotype blocks and to compare the strength of block boundaries. AB - We describe a new probabilistic method for finding haplotype blocks that is based on the use of the minimum description length (MDL) principle. We give a rigorous definition of the quality of a segmentation of a genomic region into blocks and describe a dynamic programming algorithm for finding the optimal segmentation with respect to this measure. We also describe a method for finding the probability of a block boundary for each pair of adjacent markers: this gives a tool for evaluating the significance of each block boundary. We have applied the method to the published data of Daly and colleagues. The results expose some problems that exist in the current methods for the evaluation of the significance of predicted block boundaries. Our method, MDL block finder, can be used to compare block borders in different sample sets, and we demonstrate this by applying the MDL-based method to define the block structure in chromosomes from population isolates. PMID- 12761698 TI - Outcome of hearing and vertigo after surgery for congenital perilymphatic fistula in children. AB - PURPOSE: To determine if surgery to repair a perilymphatic fistula (PLF) has any significant morbidity and to determine the outcome of the surgery in regards to hearing and vertiginous symptoms. MATERIALS AND METHODS: A retrospective study was performed with all patients undergoing PLF repair at a tertiary medical center. Each patient was assessed for comparison of their pre- and postoperative hearing levels, vertiginous complaints, and recurrences. RESULTS: One hundred sixty ears were operated on for PLF over a 13-year period. Of the 103 ears positive for PLF, 92% either stabilized or improved their hearing and 3% noticed a decrease, but this was well after surgery and not believed to be related. These results were similar in the non-PLF ears in which 95% had stabilized or improved hearing and again 3% had a much delayed decrease. Of the children who had vertiginous complaints before surgery, 91% were improved or stable. Only 1 child felt somewhat worse, but, as in the hearing loss, this was greater than 6 months after the surgery. CONCLUSIONS: Surgical repair of PLF does not result in a significant risk for postoperative hearing loss or additional vertiginous complaints. Surgical repair may prevent further deterioration of hearing loss even in those patients in whom a PLF was not identified at the time of surgery. PMID- 12761699 TI - Accuracy of computerized tomography in deep neck infections in the pediatric population. AB - OBJECTIVES: Computerized tomography (CT) is used widely to diagnose deep neck infections (DNIs), and, generally, the decision of surgical intervention is based on findings of a CT study. This study examines the accuracy of CT in differentiating abscess versus cellulitis in DNIs (lateral pharyngeal and retropharyngeal). STUDY DESIGN: This is a retrospective chart review study with re-evaluation of the CT scans by a blinded observer. METHODS: A retrospective review of medical records of 80 patients with DNIs who were evaluated with a CT study was performed. CT scans of these patients were reviewed by a radiologist who was blinded to the clinical and surgical findings and to the original CT study report. To diagnose the infection and differentiate abscess from cellulitis, our radiologist scored the CT scans regarding the following variables: low-density core, rim enhancement, soft-tissue swelling, obliterated fat planes, and mass effect. Radiologic diagnosis was compared with operative findings (whether pus found at surgery or not) in all cases treated surgically. Accuracy, sensitivity, specificity, and positive and negative predictive values of CT study were calculated. RESULTS: Thirty-nine (49%) patients were treated medically with intravenous (IV) antibiotics alone, and 41 (%51) patients were treated both surgically and medically. The overall accuracy of CT in DNI was 63%. The sensitivity, specificity, and positive and negative predictive values were 68%, 56%, 71%, and 53%, respectively. CONCLUSION: CT study has important limitations in differentiating abscess versus cellulitis in DNIs. Clinical findings as well as CT diagnosis should guide the decision of surgery. PMID- 12761700 TI - Endoscopic resection of nasopharyngeal angiofibromas by combined transnasal and transoral routes. AB - PURPOSE: To determine the role of endoscopic surgery in decreasing intraoperative bleeding, morbidity, and hospitalization period of juvenile nasopharyngeal angiofibroma resection and to describe combined endoscopic transnasal and transoral approaches. PATIENTS AND METHODS: Twelve cases of juvenile nasopharyngeal angiofibroma diagnosed by endoscopic examination, computed tomography, and angiography were selected for endoscopic resection. Tumor staging ranged from stage I(A) to II(B). Ten patients underwent preoperative selective arterial embolization, and in 1 case selective arterial ligation was used. In general, the tumors were approached endoscopically through nasal and oral cavities with 0 degrees and 30 degrees 4-mm telescopes without any incision and no packing at their termination. RESULTS: The patients were followed by endoscopy and computed tomography. There was a dramatic decrease in intraoperative bleeding and postoperative morbidity. No early postoperative complications were seen. Two recurrences were observed in 12 patients up to a mean follow-up of 15 months. CONCLUSIONS: Minimal bleeding, decreased morbidity, and shorter hospitalization period were the main reasons that prompted us to use endoscopic technique for the removal of juvenile nasopharyngeal angiofibroma. Adding transoral endoscopic approach to the transnasal endoscopic approach provides 2-sided exposure and appreciate access to angiofibroma. PMID- 12761701 TI - Aural polyps: safe or unsafe disease? AB - PURPOSE: To determine whether a case of inflammatory aural polyp constitutes a safe or unsafe disease and to arrive at the most suitable treatment option. DESIGN: Prospective study. Follow-up period of 6 months. SETTING: Hospitalized treatment in a tertiary medical college hospital that provides care for a predominantly rural population. PATIENTS: All patients treated for aural polyp, having a postoperative histopathological diagnosis of inflammatory aural polyp. Most patients (72%) belonged to the lower middle class. RESULTS: Forty-two patients treated during a 4-year-period were analyzed. Eleven cases were treated by simple aural polypectomy, of which 78% had either recurrence or persistent disease. Out of 31 patients who underwent mastoid exploration, 52% had extensive disease of the mastoid air-cell system and 35% had an underlying cholesteatoma. Six percent had persistent discharge. The disease was often associated with complications (19%). CONCLUSIONS: The presence of an aural polyp signifies well established disease of the middle ear cleft with a greater potential for complications and often obscures an underlying cholesteatoma. We propose that all cases of aural polyps should be considered as unsafe disease and subjected to a formal mastoid exploration. PMID- 12761702 TI - Autoimmunity in the etiology of otitis media with effusion. AB - Otitis media with effusion (OME) is one of the most common health problems seen in children. The role of autoimmunity in the etiology of OME has been discussed for a long time. The purpose of this investigation is to determine the role of circulating antibodies in the pathogenesis of OME. The presence of antibodies against rabbit tympanic bulla epithelium was investigated by indirect immunoflouresence in the serum samples of 25 patients with OME and serum samples of 25 healthy children as a control group. In the control group, positive staining was seen in only 1 case (4%), whereas 8 cases with OME were positively stained (%32). There was statistically significant difference between the 2 groups. The presence of antibodies against middle ear epithelium in the cases with OME suggest that autoimmunity might play a role in the formation of effusion in the tympanic cavity. PMID- 12761703 TI - The microbiology of ethmoid and maxillary sinuses in patients with chronic sinusitis. AB - PURPOSE: To investigate aerob-anaerob microorganisms growth in maxillary and ethmoid sinuses by evaluating aspiration materials from patients with chronic sinusitis. PATIENTS AND METHODS: The study was performed prospectively, and there were 31 patients (23 men, 8 women; mean age, 31.4+/-14.15, between 18-65 years) who had endoscopic sinus surgery because of chronic sinusitis. During the operation, when the maxillary sinus ostium and ethmoid sinus were opened, readily aspirated materials from sinuses were evaluated regarding aerob and anaerob bacteria. Nose and throat swap samples were collected preoperatively to determine the upper respiratory tract flora and also to understand the relationship between the flora and the microorganisms aspirated from sinuses. RESULTS: Total aerob bacteria count, which was isolated from preoperative nasal swab cultures, was 36, and aerob-anaerob bacteria count that included cultures taken from maxillary and ethmoid sinuses during the functional endoscopic sinus surgery was 42. For each 2 samples, the most common isolated aerob bacteria were coagulase (-) staphylococci. Microorganisms were isolated in 87.0% of 27 patients, in which cultures taken from maxillary and ethmoid sinuses during the functional endoscopic sinus surgery were included. It is determined that the isolated aerob bacteria rate was 90.4%, and the isolated anaerob bacteria rate was 14.2%. All of the 6 samples in which anaerob bacteria isolated were all maxillary sinus aspiration materials. Microorganisms that isolated from the nose and the sinuses were similar with the rate of 25.8%, and microorganisms that isolated from the throat cultures and sinuses were similar with the rate of 22.5%. CONCLUSIONS: This study reveals the aerob and anaerob microbiology of the maxillary and ethmoid sinuses so the treatment of chronic sinusitis will be easier. PMID- 12761704 TI - Peritonsillar and parapharyngeal space abscess in the older adult. AB - OBJECTIVE: To review the presentation and management of peritonsillar (PTA) and parapharyngeal space (PPSA) abscesses in older adults and compare this with the usual presentation and management in the younger patient. STUDY DESIGN: An 18 year retrospective review at a tertiary care hospital. METHODS: The patient database was searched by using the diagnosis codes for PTA and PPSA. The search included inpatient and outpatient visits of patients 50 years of age and older from 1983 to 2001. The charts were reviewed, and data regarding presentation, management, and outcome were recorded. Comparisons to the current literature were made. RESULTS: Fourteen patients were identified, 8 with PTA and 6 with PPSA. The most common symptoms in both groups were sore throat and dysphagia. Only 1 patient in either group was febrile. All vital signs were within normal limits in all other patients. Only 1 patient in each group experienced trismus, and no patients showed drooling. Five of the PTA patients required inpatient care, and 2 were taken to the operating room. CONCLUSIONS: PTA and PPSA are uncommon infections in the older adult, with only 14 patients identified at a tertiary care hospital over an 18-year period. Complaints of new onset sore throat and dysphagia of several days duration in patients over 50 years old should alert the evaluating physician to these 2 clinical entities, even in afebrile, nontoxic patients. PTA in this group may be more likely to require inpatient care along with surgical treatment in the operating room. PMID- 12761705 TI - Pediatric sinonasal rhabdomyosarcoma: three cases and a review of the literature. AB - OBJECTIVE: To understand the clinical presentation, management, and natural history of paranasal rhabdomyosarcoma. DESIGN: Retrospective case series review. METHODS: Retrospective medical record review of patients less than 20 years of age who presented to our facility with rhabdomyosarcoma of the nasal cavity or paranasal sinuses. RESULTS: Medical records of all pediatric patients seen in our pediatric otolaryngology clinic were reviewed from January 1, 1995, through December 31, 2000. Three patients were identified with sinonasal rhabdomyosarcoma. Their presentation, evaluation, and treatment were evaluated. Relevant literature 1966 to the present was reviewed with the assistance of Medline. CONCLUSIONS: Rhabdomyosarcoma is an aggressive pediatric malignancy, requiring a high index of suspicion to detect it in its earliest stages. Patients with suggestive symptoms should undergo a full evaluation including nasal endoscopy and imaging. Because the current chemotherapy protocols are more effective on localized disease, early diagnosis is crucial to patient survival. PMID- 12761706 TI - A case report of inverted papilloma associated with esthesioneuroblastoma treated by endoscopic sinus surgery. PMID- 12761707 TI - Nasal Rosai-Dorfman disease with intracranial involvement: a case report. AB - Sinus histiocytosis with massive lymphadenopathy (Rosai-Dorfman disease) is a rare but distinctive entity. It was initially described as a nodal-based disease but later found to affect a variety of extranodal sites. The diagnosis of Rosai Dorfman disease is based on the unique histologic changes that are characterized by proliferation of S-100 protein-positive histiocytes, the presence of emperipolesis, and a mixture of prominent lymphoplasmacytic infiltrates. The etiology of Rosai-Dorfman disease is unknown. It usually runs a benign clinic course and may resolve spontaneously, but cases running protracted courses have also been reported. We report an unusual case of recurrent extranodal Rosai Dorfman disease in nasal and paranasal cavities with intracranial involvement surviving for more than 9 years. PMID- 12761708 TI - Isolated tonsillar sarcoidosis manifested as asymmetric palantine tonsils. AB - Sarcoidosis is a chronic systemic granulomatous disease of unknown etiology. Isolated sarcoidosis of palatine tonsils in the absence of other systemic localizations is very rare, and only a few cases have been described in literature. We report a case of a 69-year-old woman presenting an asymptomatic unilateral enlargement of palatine tonsils. The pathologic report of the tonsillectomy specimen revealed the presence of noncaseating granulomatous lesions compatible with sarcoidosis. The research of acid-fast bacilli on the biopsy material using the Ziehl-Neelsen stain was negative, and no cultural growth was documented. Further investigations confirmed the diagnosis and excluded other systemic localizations of the disease. In our experience, asymmetric tonsils caused by sarcoid disease should be taken into consideration for the differential diagnosis in tonsillar enlargement. With this report, we underline the possibility of an isolated localization of sarcoidosis in palatine tonsils manifested as asymmetric enlargement, and we point out the value of histopathologic examination after tonsillectomy in selected cases. Once sarcoid lesions are found in tonsillar specimens, further investigations are mandatory to confirm the diagnosis, to detect an eventual multisystemic disease, and to exclude a possible association with malignancy. PMID- 12761709 TI - Giant-cell reparative granuloma in the temporal bone. PMID- 12761710 TI - Intraosseous hemangioma of the zygoma. AB - Zygomatic hemangioma is an extremely rare, benign, slow-growing tumor occurring mostly in adult women. The radiologic findings are diagnostic. Total excision of the tumor with primary reconstruction of the defect is the preferred treatment modality. We present a case of zygomatic hemangioma and a brief review of the literature. PMID- 12761711 TI - Rapid recovery from acoustic trauma: chicken soup, potato knish, or drug interaction? AB - OBJECTIVES: To describe the phenomenology and consider possible mechanisms mediating rapid and unexpected recovery from acoustic trauma after ingestion of a food substance (potato knish). STUDY DESIGN: Single subject with repeated test measures. SETTING: Regional Veteran's Administration Medical Center, tertiary care medical center. METHODS: Pure-tone audiometry and distortion product otoacoustic emissions (DPOAEs) performed at 6 days, 21 days, and 1 year postexposure. RESULTS: Medical treatment with corticosteriods and a diuretic alone failed to improve auditory function and related symptoms (tinnitus and aural fullness) over a 2-week period. Rapid recovery of auditory function (dramatic improvement in pure tone thresholds; reappearance of DPOAEs) and abatement of related symptoms directly followed physiologic reactions from ingesting a food substance. CONCLUSIONS: Rapid recovery from acoustic trauma was temporally correlated with urodynamic and cardiovascular reactions from ingesting food containing sulfite preservative, a substance to which the individual was allergic. Factors that may have contributed to recovery of function include massive diuresis, increased heart rate, release of biochemical mediators, mediator-induced vasodilatation, and changes in vascular or cell membrane permeability. Establishing relationships that lead to recovery of function from acoustic trauma may facilitate research and aid in the development of new treatment options for this condition. PMID- 12761712 TI - Basaloid squamous carcinoma of the larynx. AB - BACKGROUND: Basaloid squamous cell carcinoma (BSCC) is a distinct variant of squamous cell carcinoma that was first described by Wain et al in 1986. Since then, about 160 cases have been reported in the literature. Only 40 cases have been described in the larynx. METHODS: Four cases of BSCC of the larynx treated in our department between 1986 and 2000 are presented. The clinical features, biological behavior, and histopathological and immunohistochemical characteristics of this uncommon tumor are described, and the relevant literature is reviewed. RESULTS: The main clinical presentation did not differ from other histological types of laryngeal carcinoma. The clinical course, however, was much more aggressive. All the patients received aggressive therapy including radical surgery and radiation. Two patients received chemotherapy. Two of the 4 patients presented died of distant metastasis shortly after diagnosis and treatment. CONCLUSIONS: The finding of this study with a limited number of patients supports previous reports suggesting that BSCC is a highly aggressive tumor that presents in older population with male predominance. The frequency of associated regional as well as distant metastases suggests that aggressive treatment is indicated and that systemic chemotherapy should be contemplated. PMID- 12761718 TI - Genetic testing for high-risk colon cancer patients. AB - Colorectal cancer is the third leading cause of cancer-related deaths in both men and women in the United States and is estimated to have affected 148,000 people in 2002. The cumulative lifetime risk for colon cancer is approximately 5%-6%, and this risk is influenced by hereditary and lifestyle factors. In fact, 20%-30% of all colon cancer cases have a potentially definable inherited cause, and 3%-5% of colon cancers occur in genetically defined high-risk colon cancer family syndromes. Although the genes responsible for the cases of moderate-risk colon cancer remain to be characterized, many of the genes responsible for the high risk colon cancer cases have already been determined. These genetic discoveries have been translated into clinical practice and have led to improved risk assessment through the use of genetic testing. The introduction into clinical practice of genetic testing for the assessment of colon cancer risk has led to more effective management strategies for patients with potentially high-risk colon cancer and has presented new challenges to the clinician because of the unique issues involved with genetic testing. In this review, an overview of the colon cancer high-risk syndromes, with a focus on the availability and indications for genetic testing, is presented. PMID- 12761719 TI - Skin signs of gastrointestinal disease. PMID- 12761720 TI - Intestinal transplantation for gut failure. PMID- 12761721 TI - Crohn's fistula: current concepts in management. PMID- 12761722 TI - Diagnosis and management of pouchitis. PMID- 12761723 TI - Management of complications in patients receiving home parenteral nutrition. AB - Patients receiving long-term home parenteral nutrition tend to fall under the care of adult and pediatric gastroenterologists. This article reviews the management of potential infectious, mechanical and metabolic complications and describes common psychosocial issues related to the therapy. The point at which to refer the patient to an intestinal failure program offering autologous bowel reconstruction and small bowel transplantation is discussed. PMID- 12761724 TI - Postinfectious irritable bowel syndrome. AB - A small but significant subgroup of patients with irritable bowel syndrome (IBS) report a sudden onset of their IBS symptoms after a bout of gastroenteritis. Population-based surveys show that although a history of neurotic and psychologic disorders, pain-related diseases, and gastroenteritis are all risk factors for developing IBS, gastroenteritis is the most potent. More toxigenic organisms increase the risk 11-fold, as does an initial illness lasting more than 3 weeks. Hypochondriasis and adverse life events double the risk for postinfective (PI) IBS and may account for the increased proportion of women who develop this syndrome. PI-IBS is associated with modest increases in mucosal T lymphocytes and serotonin-containing enteroendocrine cells. Animal models and some preliminary human data suggest this leads to excessive serotonin release from the mucosa. Both the histologic changes and symptoms in humans may last for many years with only 40% recovering over a 6-year follow-up. Celiac disease, microscopic colitis, lactose intolerance, early stage Crohn's disease, and bile salt malabsorption should be excluded, as should colon cancer in those over the age of 45 years or in those with a positive family history. Treatment with Loperamide, low-fiber diets, and bile salt- binding therapy may help some patients. Serotonin antagonists are logical treatments but have yet to be evaluated. PMID- 12761725 TI - Fecal incontinence. PMID- 12761726 TI - Biliary imaging: a review. PMID- 12761739 TI - Exploring HBr ionization at the molecular level. PMID- 12761727 TI - Transjugular intrahepatic portosystemic shunt: current status. AB - The transjugular intrahepatic portosystemic shunt (TIPS) was developed in the 1980s for treatment of complications of portal hypertension. Once it was shown that the shunt could be placed with relative ease, TIPS was rapidly applied to the treatment of many of the complications of portal hypertension. These complications include actively bleeding gastroesophageal varices, prevention of rebleeding from varices, control of refractory cirrhotic ascites and hepatic hydrothorax, and treatment of hepatorenal failure and hepatopulmonary syndrome. TIPS has also been used as therapy for Budd-Chiari syndrome and veno-occlusive disease. Despite these broad applications, TIPS has been compared with other forms of therapy in only 2 situations: prevention of rebleeding from varices and control of refractory cirrhotic ascites. In the trials, TIPS was shown to provide better control of these 2 complications of portal hypertension than standard forms of therapy. However, there was no improvement in survival and the incidence of encephalopathy was greater for patients receiving a TIPS. Thus, the use of TIPS for the control of ascites and prevention of rebleeding from varices should be limited to a select group of patients. There have been no controlled trials for the other indications listed. Despite the apparent efficacy of TIPS in many of these situations, its use should be limited to salvage therapy pending the publication of controlled trials showing it is a better treatment than other forms of therapy. PMID- 12761728 TI - Side effects of therapy for chronic hepatitis C. PMID- 12761740 TI - Persistent triplet carbenes. PMID- 12761741 TI - Small molecules as inhibitors of cyclin-dependent kinases. AB - Cell division (mitosis) is one of the basic requirements for multicellular oranisms. The capability of a cell to replicate enables a complex assembly to be created. Faulty regulation of the control mechanism in the cell cycle leads to an excessive cell proliferation and is the cause of cancer. The key position of the cyclin-dependent kinases (CDKs) and their direct partners, as well as the fact that the majority of malign illnesses show defects in at least one of these key players of the cell cycle, is of great interest for the development of low molecular-weight CDK inhibitors. In this Review an overview of the different structural classes of ATP-competitive inhibitors of CDKs are given, whose devlopment was aimed at battling cancer. The Review shows how far the development of selective CDK inhibitors has progressed and to what extent the expectations for such drugs have so far been fulfilled. PMID- 12761742 TI - Controlling chirality and optical properties of artificial antenna systems with self-assembling porphyrins. PMID- 12761743 TI - "Catalyst analogue": a concept for constructing multicomponent asymmetric catalysts (MAC) by using a polymer support. PMID- 12761744 TI - Lithium intercalation in open-ended TiS2 nanotubes. PMID- 12761745 TI - Surface coordination chemistry: dihydrogen versus hydride complexes on RuO2(110). PMID- 12761746 TI - Template synthesis of polypyrrole nanofibers insulated within one-dimensional silicate channels: hexagonal versus lamellar for recombination of polarons into bipolarons. PMID- 12761747 TI - Generation of oligopeptides with homochiral sequences by topochemical reactions within racemic crystals of phenylalanine-N-carboxyanhydride. PMID- 12761748 TI - An unusual dynamic Fe-Hg-Pd cluster with a palladium(0) fragment stabilized by d10-d10 heterometallic bonding. PMID- 12761749 TI - Molecular encapsulation by cucurbit[7]uril of the apical 4,4'-bipyridinium residue in newkome-type dendrimers. PMID- 12761750 TI - Novel pathways for the formation of chiral binaphthyl polymers: oxidative asymmetric phenolic coupling alone and in tandem with the Glaser-Hay coupling. PMID- 12761752 TI - A Supramolecular tetra-Dawson polyoxothiometalate: [(alpha H2P2W15O56)4[Mo2O2S2(H2O)2]4[Mo4S4O4(OH)2(H2O)]2]28-. PMID- 12761751 TI - Dynamic covalent chemistry on self-templating peptides: formation of a disulfide linked beta-hairpin mimic. PMID- 12761754 TI - Furans bound face-on: sequential loss of CO in the formation of [W(CO)4(eta4-2,5 dimethylfuran)]. PMID- 12761753 TI - Metalla-azaspirophosphanes: synthesis, structure, and reactivity. PMID- 12761755 TI - Direct observation of 3D mesoporous structure by scanning electron microscopy (SEM): SBA-15 silica and CMK-5 carbon. PMID- 12761756 TI - Facile formation of N-confused porphyrin dimers by platinum(II) coordination to the outer-nitrogen atoms. PMID- 12761757 TI - Temperature-induced micelle to vesicle transition in the sodium dodecylsulfate/dodecyltriethylammonium bromide system. PMID- 12761758 TI - Synthesis, characterization, and reaction of ethynyl(phenyl)-lambda3-iodane complex with [18]crown-6. PMID- 12761761 TI - Synthesis and pharmacological investigations of some 4-hydroxycoumarin derivatives. AB - The synthesis of ten coumarin derivatives of 4-hydroxycoumarin and various unsaturated ketones and aldehydes is described. The structures of the synthesized compounds were confirmed by IR, (1)H-NMR, and mass-spectral data. Acute toxicity studies of the compounds were performed on mice by oral and intraperitoneal administration. A comparative pharmacological study of the in vivo anticoagulant effects of the derivatives with respect to warfarin, showed that the compounds have anticoagulant activity. Compounds 4-hydroxy-3-[1-phenyl-2-(4' chlorobenzoyl)ethyl]-2H-1-benzopyran-2-one 2b, 4-hydroxy-3-[1-(4-fluorophenyl)-3 oxobutyl]-2H-1-benzopyran-2-one 3a, and 3, 3'-p-bromobenzylidene-bis-(4-hydroxy 2H-1-benzopyran-2-one) 4b showed slight acute toxicity and a greater anticoagulant effect than warfarin. PMID- 12761759 TI - IBX amides: a new family of hypervalent iodine reagents. PMID- 12761762 TI - Substituted quinazolines, part 2. Synthesis and in-vitro anticancer evaluation of new 2-substituted mercapto-3H-quinazoline analogs. AB - A new series of 2-substituted mercapto-3H-quinozolines bearing 6-iodo and 2 heteroarylthio functions was synthesized and screened for their in vitro antitumor activity. Eighteen compounds were identified as active anticancer agents. N'-[(3-Benzyl-4-oxo-6-iodo-3H-quinazoline-2-yl)thioacetyl]-N(3) ethylthiosemicarbazide (10), N-benzoyl-N'-[2-(3-benzyl-4-oxo-6-iodo-3H-quinozolin 2-yl)thioacetyl]hydrazine (12), and 2-[(3, 6-dioxo-pyridazin-4-yl)thio]-3-benzyl 4-oxo-6-iodo-3H-quinazoline (20) proved to be the most active members in this study. They showed MG-MID, GI(50) values of 12.8, 11.3, and 13.8 microM, respectively. The detailed synthesis and biological screening data are reported. PMID- 12761763 TI - Modification of the structure of 4, 6-disubstituted 2-(4-alkyl-1 piperazinyl)pyridines: synthesis and their 5-HT2A receptor activity. AB - Structure-activity relationship studies of a series of novel 4, 6-disubstituted 2 (1-piperazinyl)pyridines were conducted to revise our model of serotonin 5-HT(2A) receptor antagonist. Target compounds were synthesized using the benzotriazole assisted Katritzky method. The majority of those compounds were found to be selective 5-HT(2A)/5-HT(1A) receptor ligands, though less potent than their previously described pyrimidine counterparts. In particular, the three compounds 6-8 showed the highest 5-HT(2A) receptor affinity (K(i) = 34-78 nM) and were classified as 5-HT(2A) antagonists in in vivo experiments. The influence of the structural modifications on the in vitro results was discussed; however, the elucidation of the role of the central core system requires further studies. PMID- 12761764 TI - Design and synthesis of some substituted 1H-pyrazolyl-oxazolidines or 1H pyrazolyl-thiazolidines as anti-inflammatory-antimicrobial agents. AB - Four series of 1 H-pyrazole derivatives have been synthesized. The first series was synthesized starting with the reaction of 3-(5-bromo-2-thienyl)-1-phenyl-1 H pyrazole-4-carboxaldehyde 1 with L-serine, L-cysteine, or L-penicillamine, followed by N-protection using (Boc)(2)O to provide compounds 2. The latter compounds could be N-deprotected by 4N HCl/dioxane to afford the second series 3 or reacted with NH(4)OH in the presence of DCC/HOBt to give the corresponding amides 4 followed by N-deprotection giving rise to compounds 5. The newly synthesized compounds were evaluated for their anti-inflammatory-antimicrobial activities. In addition, the ulcerogenic and acute toxicity profiles were determined. Compound 5b (2RS, 4R)-2-[3-(5-bromo-2-thienyl)-1-phenyl-1H-pyrazol-4 yl]-5-methylthiazolidine-4-carboxamide, proved to be the most active anti inflammatory-antimicrobial agent in the present study with a good safety margin and no ulcerogenic effect. PMID- 12761765 TI - Activities of prenylphenol derivatives from fruitbodies of Albatrellus spp. on the human and rat vanilloid receptor 1 (VR1) and characterisation of the novel natural product, confluentin. AB - Several prenylphenols from basidiocarps of European and Chinese Albatrellus spp., namely grifolin (1), neogrifolin (2), confluentin (3), scutigeral (4), and albaconol (5) were investigated concerning their activities in test models for vanilloid receptor modulation. The isolation of these compounds from A. confluens and structure elucidation of the novel natural product confluentin (3) are described. The effects of scutigeral and neogrifolin on vanilloid receptors were studied by means of electrophysiological methodology on rat dorsal root ganglion neurons as well as on recombinant cell lines expressing the rat VR1 receptor. Concurrently, the effects of compounds 1-5 on a reporter cell line expressing the human vanilloid receptor VR1 were measured. In contrast to previous studies reported in the literature, the results of these investigations suggest that fungal prenylphenols act as weak antagonists (activity in the microM range), rather than exhibiting agonistic activities. PMID- 12761766 TI - The structure and ion channel activity of 6-benzylamino-3-hydroxyhexa cyclo[6.5.0.0(3,7).0(4,12).0(5,10).0(9,13]tridecane. AB - A novel compound, 6-benzylamino-3-hydroxyhexacyclo [6.5.0.0(3, 7).0(4, 12).0(5, 10).0(9, 13)]-tridecane, was synthesized as part of an ongoing study to explore the ion channel activity of polycyclic cage amines. The known polycyclic calcium channel antagonist, 8-benzylamino-8, 11-oxapentacyclo [5.4.0.0(2, 6).0(3, 10).0(5, 9)]undecane (NGP 1-01) served as the lead compound and as a positive control for channel activity. The title compound inhibited calcium currents at test concentrations of 10 microM at depolarized membrane potentials (in the potential range where the L-type calcium channel inactivates). At the test concentrations modulating effects were also observed for sodium and the delayed rectifier potassium currents. Due to its activity at both Ca(2+) and Na(+) channels, this compound may offer utility as a cardiovascular and/or neuroprotecting agent. PMID- 12761767 TI - Microflora involved in textile dye waste removal. AB - Textile dyes are heavily used in factories for coloring different cloth materials. This work was designed to identify microorganisms capable of removing textile dyes, either by biodegradation or by biosorption. We expected to isolate microorganisms adapted to high dye concentrations from sites near textile industry complex. An experiment was conducted to study the efficiency of the isolates in removing textile dyes. The tested dyes were used as carbon and nitrogen sources for isolation of soil and/or water microorganisms capable of removing textile dyes wastes from factories effluent. The results indicated the low efficiency of both bacteria and actinomycetes in clean-up the effluent from the waste dyes in 10-21 days. On the other hand six fungal isolates were obtained by plating factory effluent on Martin's medium and media containing dyes as the sole source of carbon and nitrogen for growth. These isolates fell in two genera, Aspergillus and Trichoderma. Results of these studies revealed the potential capacity of these fungi to decolorize the tested dyes in comparatively short time (2-24 hours) indicating strong efficiency of dye bioremediation by the fungal isolates. Since the process involved is mostly fast interaction between the fungal mycelium and the dye in the media, the possible mechanism could be based on a biosorption of such chemicals on the intact fungal biomass, rather than direct biodegradation of the compounds. PMID- 12761768 TI - Distribution of zoosporic fungi in the mud of major Egyptian lakes. AB - Twenty-six identified and three unidentified species belonging to eight genera of zoosporic fungi were recovered from seventy-six mud samples of Egyptian lakes using sesame seeds as baits. Nineteen samples were collected along the borders of four major lakes. Pythium, Phytophthora and Allomyces, yielded the highest spectra of zoosporic species, while Aqualinderella, Blastocladia and Dictyuchus, had one species. Pythium, Aphanomyces, Allomyces, Aqualinderella, Phytophthora and Saprolegnia were the dominant zoosporic genera while Blastocladia and Dictyuchus were rare. The most prevalent species of zoosporic fungi were Aqualinderella fermentans, Aphanomyces laevis and unidentified Pythium species while the other isolated species in this study were rare. Menzalah was the richest lake with zoosporic species followed by Borolous and Karoon. Some zoosporic fungal genera were recovered from the mud of the four tested lakes (participator genera) and others from three lakes or from only two lakes. Physicochemical characteristics of the mud samples (pH, total soluble salts and organic matter) revealed no correlation with the fungal occurrence and distribution in these lakes. PMID- 12761769 TI - Occurrence of yeasts in municipal wastes and their behaviour in presence of cadmium, copper and zinc. AB - Seven yeasts strains have been isolated from sewage sludge. Also six samples of compost with different sieving, composting times and origins, have been analysed. Apparently, composting processes negatively affect the viability of yeasts, as none could be isolated from the compost samples. The margins of tolerance of the yeasts to Cd, Cu and Zn have been determined. The physiological response to metals was similar in all the species studied, and in general, kinetic parameters (mu and lag) were affected. Metal uptake ability was also studied and inter- and intra-specific heterogeneity was detected, thus indicating that both the tolerance to metals and the capacity of the uptake were dependent on ionic metal and yeast species. The effect of the presence of multi-metal ions on the uptake capacity of each individual metal was assayed for two selected yeasts, Pichia guilliermondii and Torulaspora delbrueckii. The uptake of each individual metal varied with the combination assayed, and when both strains were compared different results were also found. PMID- 12761770 TI - Evaluation of SDS-polyacrylamide gel systems for the study of outer membrane protein profiles of clinical strains of Acinetobacter baumannii. AB - The outer membrane protein (OMP) profiles of 23 blood isolates of Acinetobacter baumannii representing all the different antimicrobial susceptibility patterns observed during a 3-year period in a Spanish hospital were studied. OMPs extracted from envelopes of sonicated cells after solubilisation with 2% of N lauryl-sarcosinate were analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) using the Laemmli's buffers. Eight running gel systems differing in the concentration of polyacrylamide (8%, 10% and 12%) and in the absence or presence of urea (4 M and 6 M) were used in a preliminary study analysing the OMP profiles of four clonally unrelated strains of A. baumannii. When this study was completed, the OMPs of the 23 A. baumannii were analysed in 10% SDS-polyacrylamide gels with 6 M urea and in 12% SDS-polyacrylamide gels. Ten OMP profiles were observed in 10% SDS-polyacrylamide gels with 6 M urea, whereas only 5 OMP profiles were visualised using 12% SDS-polyacrylamide gels. The OMP profiles obtained in 10% SDS-polyacrylamide gels with 6 M urea only partially correlated with those observed in 12% SDS-polyacrylamide gels. In conclusion, the use of 10% SDS-polyacrylamide gels with 6 M urea is recommended for the study of OMP profiles of A. baumannii. PMID- 12761772 TI - Effect of carbon source on alkaline phosphatase production and excretion in Aspergillus caespitosus. AB - The effect of several carbon sources on the production of alkaline phosphatase by the thermotolerant Aspergillus caespitosus was analysed. The fungus released high levels of alkaline phosphatases into the medium after being cultured for long periods with xylan or industrial residues such as wheat raw and sugar cane bagasse in the culture media. In contrast, the alkaline phosphatase activities were found only intracellulary when the fungus was cultured in glucose supplemented media. The pH of the medium likely affects the process of enzyme secretion according to the carbon source used. Addition of xylan or industrial residues in the culture medium stimulated the secretion of phosphatases. In contrast, media supplemented with glucose or disaccharides promoted retention of these enzymes into the cells. The subcellular location activities of alkaline phosphatases were studied using histochemical and immunochemical methods and showed that alkaline phosphatases were present in the mycelial walls and septa. PMID- 12761771 TI - Molecular diversity of mesophilic and thermophilic bacteria in a membrane bioreactor determined by fluorescent in situ hybridization with mxaF- and rRNA targeted probes. AB - An evaluation of the efficiency of treatment of kraft mill foul condensates in a membrane bioreactor was carried out in the laboratory. Efficiency and rate of methanol removal were quantified at operating temperatures of 35, 45 and 55 degrees C. The structure of the bacterial community present in the reactor biomass at the different operating temperatures was evaluated by in situ hybridization of the biomass samples with fluorescently-labelled probes (FISH) targeting the Eubacteria, the alpha, beta and gamma subclasses of the Proteobacteria, the low G + C content Gram-positive bacteria (Bacillus spp.), while community function was evaluated by in situ hybridization with a methanol dehydrogenase gene (mxaF) probe. Methanol removal efficiency decreased from 99.4 to 92%, and removal rate from 2.69 mg MeOH/l x min to 2.49 mg MeOH/l x min when the operating temperature was increased from 35 to 55 degrees C. This decrease in methanol removal was accompanied by a decrease (from 58% to 42%) in the relative proportion of cells that hybridized with the mxaF probe. The relative proportion of Bacillus spp. increased from 5 to 20% while the proportion of members of the alpha subclass of Proteobacteria decreased from 16% to 6% when the bioreactor operating temperature was raised from 35 to 55 degrees C. The relative proportions of bacteria belonging to the beta (22-25%) and gamma (18-20%) subclasses of the Proteobacteria remained relatively constant regardless of operating temperature. Proteobacteria (alpha, beta and gamma subclasses) and Bacillus spp. represented 61, 67 and 71% of the Eubacteria in the biomass sampled at 35, 45 and 55 degrees C, respectively. The FISH technique was shown to be an efficient method for detection of both structural and functional changes in the bacterial communities that could be related to efficiency of methanol removal in a membrane bioreactor operating at different temperatures. PMID- 12761773 TI - Evaluation of biotoxicity of textile dyes using two bioassays. AB - The toxicity of eight textile dyes was evaluated using two bioassays namely: Ames test and seed germination test. The Ames test is widely used for the evaluation of hazardous mutagenic effect of different chemicals, as a short-term screening test for environmental impact assessment. The eight-textile dyes and Eithidium bromide dye (as positive control) were tested with five "his" Salmonella typhimurium strains: TA 100; TA 98; TA 1535; TA 1537; TA 1538. Using six concentrations of each dye (2.5 microg/ml, 4.5 microg/ml, 9 microg/ml, 13.5 microg/ml, 18 microg/ml, and 22.5 microg/ml) revealed that, most of the dyes were mutagenic for the test strains used in this study. The high concentrations of dye eliminated microbial colonies due to the high frequency of mutation causing lethal effect on the cells. In this work the phytotoxicity of different soluble textile dyes was estimated by measuring the relative changes in seed germination of four plants: clover, wheat, tomato and lettuce. The changes in shooting percentages and root length as affected by dye were also measured. Seed germination percent and shoot growth as well as root length were recorded after 6 days of exposure to different concentrations of textile dyes in irrigation water. The results show that high concentrations of dyes were more toxic to seed germination as compared with the lower concentrations. However, the low concentrations of the tested dyes adversely affected the shooting percent significantly. PMID- 12761774 TI - Biomass and exopolysaccharide production in submerged cultures of Pleurotus ostreatoroseus Sing. and Pleurotus ostreatus "florida" (Jack.: Fr.) Kummer. AB - The species of the genus Pleurotus produce large amounts of biomass and exopolysaccharide (EPS) in submerged cultures, which may be used for biotechnological purposes. In the present work two Brazilian autochthonous strains of edible Pleurotus (P. ostratoroseus Sing. and P. ostreatus "florida") were used. The fungi grown in liquid Potato Dextrose medium (PD) were used as inocula to cultivate those microorganisms in the POL culture medium. After a 9 day incubation, the optimal growth time for biomass production, P. ostreatus "florida" presented higher biomass production (22.8 g d.w./l) than P. ostreatoroseus (16.8 g d.w./l). After a 7-day incubation, the optimal growth time for EPS production, P. ostreatoroseus produced higher amounts of crude EPS (5.8 g d.w./l) than P. ostreatus "florida" (1.4 g d.w./l). Relative carbohydrate composition for P. ostreatoroseus and P. ostreatus "florida" EPS were: glucose (95.5-87.7), galactose (traces - 4.9), mannose (traces - 3.1), xylose (1.3-2.5), and arabinose (3.2-1.8). Lower ammonium sulfate concentration in the POL culture medium increased the exopolysaccharides production by P. ostreatoroseus. PMID- 12761775 TI - Phenol degradation by a Graphium sp. FIB4 isolated from industrial effluents. AB - In this work, we show that the fungal strain Graphium sp. FIB4 was able to use phenol as the sole carbon source. Higher degradation of phenol was accomplished by alginate-immobilized mycelial mass than by mycelial suspensions of Graphium sp. FIB4. Free mycelium exhibited higher degradation rates when compared with the alginate-immobilized mycelium in the presence of 14 mM of phenol or less. Above this concentration, degradation rates by free mycelium decreased and the immobilized mycelium showed higher values. The maximum degradation rate for 8 mM phenol was found to be 20.13 mg/l x h by free mycelia and 16.24 mg/l x h by immobilized mycelial mass in the presence of 18 mM phenol. When the fungus was grown on medium without phenol, catechol 1,2-dioxygenase activity was not detected. This enzyme activity was induced at phenol concentrations as low as 0.05 mM and up to 6 mM at 24 h incubation at 30 degrees C, suggesting that catechol was oxidized by the ortho type of ring fission. Addition of glucose reduced phenol consumption rate, and both substrates were used simultaneously. Glucose concentrations higher than 0.075% repressed the induction of phenol oxidation by Graphium sp. FIB4 grown on glucose. But glucose did not fully repress utilization of phenol by phenol-pre-induced cells. Immobilization and addition of calcium and barium ions were detrimental to the stability of catechol 1,2-dioxygenase activity and phenol degradation by Graphium sp. FIB4. PMID- 12761776 TI - Nucleotide sequence of MPB63 gene in Mycobacterium bovis BCG Tokyo. AB - Mycobacterium bovis BCG has been used for the prevention of tuberculosis and as therapy for bladder tumor. MPB63 in M. bovis BCG is one of the immunogenic proteins and is secreted in large quantities. Therefore, it is of interest that the MPB63 gene be examined for the determination of its nucleotide sequence. A fragment of 820 base pairs (bp) including the MPB63 gene was prepared by amplification using polymerase chain reaction (PCR) employing M. bovis BCG Tokyo chromosomal DNA as a template and its nucleotide sequence was determined. The nucleotide sequence of mpb63 in M. bovis BCG was then compared with that of mpt63 in M. tuberculosis. The result indicated that the nucleotide sequences between two protein genes were quite agreeable in the genes' structural and upstream regions, except that one base change from C in mpt63 to G in mpb63 was detected in the downstream trailer sequence. This suggests that the genetic information of M. bovis BCG is not entirely identical to that of M. tuberculosis, although the characteristics of both microorganism are very similar to each other. PMID- 12761779 TI - The principles of migration and dispersion in capillary zone electrophoresis in nonaqueous solvents. AB - Nonaqueous solvents are interesting media for capillary zone electrophoresis as they can affect all relevant parameters governing the separation of sample zones. However, for a rational planning of the working conditions and an appropriate interpretation of the results obtained, the basic principles of ion migration and zone dispersion must be understood. Many solvent induced effects need to be carefully considered and recognized before full exploitation of nonaqueous solvents can take place. It is the goal of this overview to present the fundamental physicochemical aspects of capillary zone electrophoresis in nonaqueous solvent systems. Therefore, the detailed discussion is related to the effect of organic solvents on electrophoretic mobilities (based on the theory of conductance), acid-base dissociation behavior (based on the transfer activity coefficient and medium effect), pH, separation efficiency (with regard to mobility and diffusion coefficient in dilute solutions), resolution, and electroosmotic flow. PMID- 12761780 TI - Effects of background electrolyte composition and addition of selectors on separation selectivity in nonaqueous capillary electrophoresis. AB - This review gives a survey of the approaches employed to obtain, enhance and tune selectivity in nonaqueous capillary electrophoresis (NACE). Recent developments in NACE are described and the effects of background electrolyte composition and addition of selectors on separation selectivity are discussed. The use of one organic solvent, a mixture of several organic solvents or the use of additives to tune separation selectivity in NACE is presented and a list of relevant applications is included. PMID- 12761781 TI - Effects of organic solvents on sample pretreatment and separation performances in capillary electrophoresis. AB - A review of recent developments in theoretical as well as application studies concerning the use of organic solvents, either as purely nonaqueous solvents, hydro-organic mixtures, or a combination of an organic solvent with another organic modifier(s), in the sample matrix and/or separation buffer for effecting sample pretreatment and/or improving separation performances in capillary electrophoresis (CE) is presented. In particular, recent advances made in furthering the basic understanding of selectivity changes that occur in capillary zone electrophoresis due the presence of organic solvents in the separation medium, based on in-depth studies of fundamental processes, such as acid-base chemistry, ion-ion and ion-solvent interactions, were discussed in detail. The utilization of organic solvents for improving the resolution of highly challenging and important separations, i.e., those involving the separation of positional and optical isomers, was also critically reviewed. Furthermore, a comprehensive survey of the use of organic solvents for on-line sample pretreatment, e.g., minimizing aggregation and maximizing solubilization of hydrophobic analytes, improving concentration detection sensitivity for analytes via the use of sample stacking, was presented and discussed. Moreover, recent applications involving the use of organic solvents for improving the CE separations of a variety of molecular species with significance in various disciplines, including biological, environmental and pharmaceutical areas, were summarized and tabulated. PMID- 12761782 TI - The role of organic solvents in the separation of nonionic compounds by capillary electrophoresis. AB - Although nonionic compounds can be separated by micellar electrokinetic chromatography (MEKC), application of this technique is restricted by a somewhat limited elution range. Incorporation of organic solvents in the background electrolyte (BGE) greatly extends the scope of MEKC and provides a major variable in optimizing the separation of neutral analytes. This paper provides a systematic review of the principles and scope of the separation of neutral analytes by capillary electrophoresis (CE) in organic-aqueous solution. The methods surveyed include those that use tetraalkylammonium salts, dioctyl sulfosuccinate, lauryl poly(oxyethylene) sulfate. Polyaromatic hydrocarbon (PAH) compounds can be separated using sodium hexadecyl sulfate in 70% methanol (30% aqueous) to 100% methanol. PMID- 12761783 TI - Solvent effects in microemulsion electrokinetic chromatography. AB - The contribution of organic solvents to the mechanisms responsible for separation in microemulsion electrokinetic chromatography (MEEKC) is reviewed. Organic solvents are needed as constituents of microemulsions for a series of reasons. (i). A water-immiscible organic substance is used to form the actual oil phase of the microemulsion, (ii). a less hydrophobic solvent is commonly employed as a so called co-surfactant, and (iii). in many cases an organic modifier is added to influence the solubility of the analytes in the aqueous phase of the microemulsion. All these organic solvents do not only participate in the separation in their actual function, but also interact with each other and the analytes. Variations in separation selectivities triggered by changes in the nature and/or concentration of these organic solvents present in microemulsions suitable for MEEKC are discussed in this work. PMID- 12761784 TI - Capillary zone electrophoresis in methanol: migration behavior and background electrolytes. AB - Nonaqueous (NA) solutions are often used as background electrolytes (BGEs) and NA solvents are added to aqueous BGEs as organic modifiers in capillary zone electrophoresis (CZE), in order to optimize the separations. This can be tricky, however, because the pH* and pK* concepts may be totally different in NA solvents, whereas often less knowledge is available concerning phenomena, such as system zones, applying NA solvents. In this paper, the concepts of pH* and pK* are considered for methanol as a solvent and pK* values are determined for several components in mixtures of water and methanol. With a mathematical model, adapted for calculations in methanol, parameters are calculated describing the fronting or tailing character of peaks and the question of peaks or dips, and the existence of system zones is discussed for pure methanol as a solvent. These aspects are experimentally verified, applying BGEs useful for the separation of cationic species in the indirect UV mode. It can be concluded that the mathematical model developed for aqueous BGEs is applicable to BGEs in methanol, too, and that the behavior of BGEs in methanol is comparable with that in water concerning the fronting or tailing character of peaks and the question of peaks and dips, although the mobilities and pK values can change significantly. PMID- 12761785 TI - Influence of solvent on temperature and thermal peak broadening in capillary zone electrophoresis. AB - The present paper deals with the role of the solvent on thermal peak broadening. One main solvent property that determines the magnitude of the temperature gradient due to the generation of Joule heat in capillary zone electrophoresis is the thermal conductivity. As organic solvents have lower thermal conductivity than water (methanol and acetonitrile, e.g., nearly by a factor of 3) it can be hypothesized that the temperature gradient inside the capillary is more pronounced in organic solvents compared to an aqueous solution. On the other hand, the temperature dependence of the ion mobility (which is responsible for the velocity profile and thus for thermal peak broadening) is smaller in organic solvents. To get insight into the thermal effect of the solvent, first the temperature of a solution in a cylindrical tube was calculated utilizing the heat balance equation. It was shown that the two theoretical models most common in the literature (based on the analytical solution or on an assumption of the parabolic temperature profile in the tube, respectively) give the same results. The latter model was chosen for the further calculations, adding a quadratic term to express the electric conductivity as a function of the temperature. The temperature at the inner capillary wall and center as function of the capillary dimensions and the electric power was computed for electrolytes with a given conductivity at 25.0 degrees C with water, methanol, and acetonitrile as solvents. Capillary cooling systems used were circulating liquid cooling, enforced air-cooling, and natural convection in still air. The mean temperature (averaged over the cross section) resulting from Joule heating was compared with experimentally determined temperatures established upon application of an electric field; the latter temperature was derived from the measurement of the electric conductance of the background electrolyte solution and its (measured) temperature dependence. All investigations were carried out with solutions of the same initial electric conductivity (about 0.5 S.m(-1) at 25.0 degrees C). Agreement is found for natural convection conditions, and the deviation between theoretical and experimental results for the forced air and circulated liquid cooling systems can be related to the poorly defined thermal conditions of the capillaries in commercial instrumentation (with a part in a thermostated cassette and a part outside). For given conditions the temperature gradients in the organic solvents exceed largely those in water, independent of the type of cooling. As a consequence, the thermal plate height is significantly larger in organic solvents, at least under conditions where the deviation from the Nernst-Einstein limiting case is not too high. However, even for the maximum applicable field strengths the thermal plate height contributions are negligible compared to longitudinal diffusion in all solvents. PMID- 12761786 TI - Peak dispersion and contributions to plate height in nonaqueous capillary electrophoresis at high electric field strengths: propanol as background electrolyte solvent. AB - Peak dispersion effects in nonaqueous capillary electrophoretic separations of aromatic anionic analytes were investigated in a propanolic background electrolyte solution. Poly(glycidylmethacrylate-co-N-vinylpyrrolidone) coating was applied to the capillary to suppress the electroosmotic flow and to improve the repeatability of the migration times. Electrical field strengths up to 2000 Vcm(-1) were applied in separations and the separation efficiencies were compared with theoretical values calculated on the basis of plate height theory. The contributions to the total plate height were calculated for injection plug length, diffusion, Joule heating, electromigration dispersion, analyte adsorption to the capillary wall, and detector slit aperture length. Analyte diffusion coefficients were measured by Taylor dispersion method, while distribution constants were measured chromatographically. Agreement between the calculated and empirical results was fairly good even though some approximations were required. In most cases the longitudinal diffusion contribution governed the total plate height, while the contribution of Joule heating was insignificant even at exceptionally high field strengths used. The relatively long detection slit aperture was found to influence the separation efficiency strongly, while the other dispersion sources that were investigated were of minor importance, except for adsorption in the case of one analyte. With all analytes, the dispersive effect of longitudinal diffusion was reduced as the field strength was increased, leading to enhanced migration velocities and faster separations. PMID- 12761787 TI - Influence of electrolyte nature on the separation selectivity of amphetamines in nonaqueous capillary electrophoresis: protonation degree versus ion pairing effects. AB - The simultaneous analysis of Ecstasy and its derivatives in an acetonitrile methanol (80:20 v/v) mixture was previously shown to be strongly dependent on the nature of the electrolyte (acetate versus formate). To elucidate the phenomena involved, systematic experiments were conducted in this solvent medium. Conductivity measurements allowed to evaluate the ion-pairing rate in the background electrolyte (BGE) and thereby distinguish between electrolyte concentration and ionic strength. The influence of electrolyte concentration on analyte effective mobilities micro(eff)) was studied by capillary electrophoresis (CE). As micro(eff) extrapolated to infinite dilution proved to be independent of the nature of the electrolyte, selectivity changes could not be attributed to a modification in the protonation degree of amphetamines. Experimental mobility data were then confronted to existing theoretical mobility models to discriminate between ion pairing or simple ionic strength effect. Ion-pair formation in a BGE containing acetate was highlighted with an ion-pairing model and ion-pair formation constants between each amphetamine and acetate ion were calculated. PMID- 12761788 TI - Modeling the zeta potential of silica capillaries in relation to the background electrolyte composition. AB - A theoretical relation between the zeta potential of silica capillaries and the composition of the background electrolyte (BGE) is presented in order to be used in capillary zone electrophoresis (CZE). This relation is derived on the basis of the Poisson-Boltzmann equation and considering the equilibrium dissociation of silanol groups at the capillary wall as the mechanism of charge generation. The resulting model involves the relevant physicochemical parameters of the BGE capillary interface. Special attention is paid to the characterization of the BGE, which can be either salt or/and buffer solutions. The model is successfully applied to electroosmotic flow (EOF) experimental data of different aqueous solutions, covering a wide range of pH and ionic strength. Numerical predictions are also presented showing the capability of the model to quantify the EOF, the control of which is relevant to improve analyte separation performance in CZE. PMID- 12761789 TI - Comparison of different algorithms to calculate electrophoretic mobility of analytes as a function of binary solvent composition. AB - Ten different mathematical models representing the electrophoretic mobility of analytes in capillary electrophoresis in mixed solvents of different composition have been compared using 32 experimental data sets. The solvents are binary mixtures of water-methanol, water-ethanol and methanol-ethanol, respectively. Mean percentage deviation (MPD), overall MPD (OMPD) and individual percentage deviation (IPD) have been considered as comparison criteria. The results showed that a reorganized solution model, namely the combined nearly ideal binary solvent/Redlich-Kister equation, is the most accurate model among other similar models concerning both correlation ability and prediction capability. PMID- 12761790 TI - Dual stacking of unbuffered saline samples, transient isotachophoresis plus induced pH junction focusing. AB - A dual stacking mechanism based on transient isotachophoresis (TITP) and induced pH junction focusing is demonstrated as a means to increase the concentration sensitivity in capillary electrophoresis of highly saline samples. When stacking was carried out with an unbuffered saline sample of fluorescein between two zones of low mobility background electrolyte at high pH under an electric field of reverse polarity, two transient peaks at both boundaries of the sample zone were observed. One peak at the rear boundary could be inferred as a transient isotachophoretic stacked zone. Through computer simulations of an unbuffered sample with a high concentration of sodium chloride, we showed that the fast moving zones of sodium and chloride ions induced pH changes at both boundaries to satisfy the electroneutrality condition and that the peak at the front boundary was due to the induced pH junction. To verify the pH changes, an indicator, thymol blue, was added to an NaCl solution and the color changes under an electric field were observed. The proposed mechanism was supported by observing the dual stacking procedure for an unbuffered sample of 4-nitrophenol and measuring additional sensitivity enhancements by dual stacking for ten weakly acidic compounds. For the ten analytes including nucleoside phosphates, every dual stacking of an unbuffered sample exhibited an additional enhancement up to 86% larger than that of usual transient isotachophoresis of the corresponding buffered sample without loss of separation efficiency and reproducibility. Therefore, it would be useful to skip over buffering in sample preparation for TITP, contrary to the general recommendation. PMID- 12761792 TI - Salt dependence on vancomycin-herbicide enantiomer binding: capillary electrophoresis study and theoretical approach. AB - In a previous paper, a mathematical model was developed for the estimation of binding constants by capillary electrophoresis (Guillaume and Peyrin, Anal. Chem. 1999, 71, 2046-2052). This model was applied to determinate the binding constant of the guest-vancomycin complex as well as the degree of complexation n(c) (percent of complexed guest). In this work, the guest molecule were a series of acid herbicide enantiomers (aryloxypropionic, aryloxyphenoxypropionic, and aminopropionic acid). The effects of the run buffer salt concentration on the herbicide-vancomycin binding was clearly described by the determination of specific physicochemical parameters as well as thermodynamic data. These results demonstrate that a better understanding of the interactions involved in the guest molecule-vancomycin binding allows to optimize the enantiomeric separation of this chiral guest molecule. PMID- 12761791 TI - Capillary electrophoretic study of the binding of zinc(II) ion to bacitracin A1 in water-2,2,2-trifluoroethanol. AB - Binding of Zn(2+) to bacitracin A(1) was studied by capillary electrophoresis in water/2,2,2-trifluoroethanol (70/30 v/v) at different apparent pH values in order to estimate the association constant of metal, the acidic dissociation constants and the Stokes radii of both free and bounded peptide in apolar environment. The Stokes radii of the free peptide species were compared with those in aqueous solution, as obtained in a recent study performed by our group, indicating that apolar environment stabilizes bacitracin A(1) in a conformational structure with the lateral chain of apolar amino acids exposed on the external surface. This conformation of the macrocyclic dodecapeptide is ready to interact with Zn(2+) ion, as pointed out by the strong increase of the association constant measured in water/2,2,2-trifluoroethanol with respect to the value obtained in aqueous solution. In addition, whereas Zn(2+) ion binding in aqueous solution provides a sensible reduction of peptide Stokes radius, no sensible variations following to ion binding were observed in hydro-organic solution. The present results suggest that the apolar environment, rather than the metal ion binding, could be responsible for the conformational transition that brings bacitracin A(1) towards its biologically active structure.* PMID- 12761793 TI - Heteroconjugation-based capillary electrophoretic separation of phenolic compounds in acetonitrile and propylene carbonate. AB - A mixture of methyl- and hydroxy-substituted phenols was separated by capillary electrophoresis in pure acetonitrile and propylene carbonate. Interactions between undissociated phenolic compounds and the background electrolytes were investigated. In the present work, benzyltriethylammonium chloride, tetrabutylammonium acetate, and two room temperature-molten salts, 1-butyl-3 methyl imidazolium trifluoroacetate and 1-butyl-3-methyl imidazolium heptafluorobutanoate, were used as background electrolytes. The formation of a negative complex between background electrolyte anion and neutral phenolic compound was observed and the formation constant calculated. The formation constants for anion-analyte complexes were approximately the same in propylene carbonate and in acetonitrile. In both solvents the formation constants were the highest for acetate and the lowest for trifluoroacetate. The separation of analytes was slightly influenced by the nature of the solvent: in acetonitrile the resolution between peaks was higher for 1,3-dihydroxyphenol and 1,3,5 trihydroxyphenol, in propylene carbonate 3-methylphenol and phenol were better separated. It was demonstrated that traces of water influence the mobilities of anion-phenol complexes in propylene carbonate. PMID- 12761794 TI - Electrophoretic analysis of quinone anion radicals in acetonitrile solutions using an on-line radical generator. AB - We have investigated analysis of anion radicals of phenanthrenequinone (PhQ) and anthraquinone (AQ) using acetonitrile-capillary electrophoresis (CE) under anaerobic conditions. PhQ and AQ have relatively high negative reduction potentials meaning that their anion radicals are re-oxidized quite readily by the surrounding O(2) to disappear during analysis and we failed to detect them with our previous system. In this work, we have developed an on-line system combining a unique electrolysis cell for generation of the radicals and a CE unit to keep the analysis system free from external O(2) molecules and to reduce analysis time remarkably. As a result, electrophoretic detection of the anion radicals of PhQ and AQ has been achieved. Furthermore, we have observed hydrogen-bonding interaction between the anion radicals and dimethylurea (DMU) using the present system and have indicated a characteristic interaction of the anion radical of PhQ as an ortho-quinone with DMU. PMID- 12761795 TI - Method development for the analysis of trans-fatty acids in hydrogenated oils by capillary electrophoresis. AB - A novel capillary electrophoresis methodology using UV indirect detection (224 nm) for the analysis of trans-fatty acids in hydrogenated oils was proposed. The electrolyte consisted of a pH 7 phosphate buffer at 15 mmol x L(-1) concentration containing 4 mmol.L(-1) sodium dodecylbenzenesulfonate, 10 mmol x L(-1) polyoxyethylene 23 lauryl ether (Brij 35), 2% 1-octanol and 45% acetonitrile. Under the optimized conditions, ten fatty acids, C12:0, C13:0 (internal standard), C14:0, C16:0, C18:0, C18:1c, C18:1t, C18:2cc, C18:2tt and C18:3ccc were baseline-separated in less than 12 min. The proposed methodology was applied to monitor the formation of trans-fatty acids during hydrogenation of Brazilnut oil. A crude oil sample (42.1% linoleic acid, 37.3% oleic acid, 13.4% palmitic acid, and 7.0% stearic acid) was mixed with 0.25% of a nickel-based catalyst and submitted to two independent hydrogenation conditions: 175 degrees C, 3 atm, 545 rpm for 60 min (GH(1) sample), and 150 degrees C, 1 atm, 545 rpm for 30 min (GH(2) sample). For the most severe hydrogenation condition (higher temperature and pressure, under longer reactional period), a more complete conversion of linoleic and oleic acids into stearic acid occurred with concomitant formation of the trans-species, elaidic acid (C18:1t). For the milder hydrogenation procedure that generated sample GH(2), larger amounts of linoleic and oleic acids remained, in addition to the transformations already observed in the GH(1) sample. PMID- 12761796 TI - Separation of (Z)- and (E)-isomers of thioxanthene and dibenz[b,e]oxepin derivatives with calixarenes and resorcinarenes as additives in nonaqueous capillary electrophoresis. AB - Five acidic calix[4]arenes with carboxylic or sulfonic groups at either the upper or lower rim of the cavity and one resorc[4]arene were investigated to separate three thioxanthenes (flupentixol, clopenthixol, chlorprothixene) and a dibenz[b,e]oxepin derivative (doxepin) with cis-/trans-isomerism by nonaqueous capillary electrophoresis (NACE). Partial filling of the capillary with the UV absorbing selectors led to a low detection limit and an advantageous signal-to noise ratio (S/N). A sufficient electrophoretic mobility of the calixarenes towards the anode was necessary to outweigh the oppositely directed electroosmotic flow (EOF). This depended from the functional groups, the dissociation and the hydrodynamic radius of the cyclophanes. In contrast, the resorcinarene was useable only by addition of sodium dodecyl sulfate (SDS) because only the complex of the two selectors had an anodic apparent electrophoretic mobility. p-Sulfonyl-calix[4]arene (ss-a1) was the most capable selector for all E/Z-isomers with maximal alpha-values ranging from 1.056 for doxepin to 1.224 for chlorprothixene. This was due to the sufficient migration in reversed direction to the EOF even at low pH* values of 3.0. Otherwise, electrostatic as well as hydrophobic interactions with the positively charged isomers seem to contribute to a superior recognition. Increasing the concentration up to 15 mM ss-a1 and using acidic media (pH* 5.0) led to high separation efficiency. Changing the organic solvent provides a powerful tool to improve selectivity with N,N-dimethylformamide-methanol (DMF-MeOH)-mixtures for thioxanthenes. Further electrophoretic parameters were optimized, such as the concentration of the electrolytes, the addition of SDS, the kind of electrolytes and the voltage. Distinct differences in selectivities were found between the derivatives with thioxanthene and dibenzo[b,e]oxepin ring system, respectively. Further, the different basic side chain was responsible for the different selectivity at higher pH* values. In contrast, the substitution at position 2 of the thioxanthenes played a secondary role. Based on the studies of single parameters a method for a simultaneous separation of the four pairs of isomers within 13 min was developed. PMID- 12761797 TI - Microemulsion electrokinetic chromatography for the analysis of green tea catechins: effect of the cosurfactant on the separation selectivity. AB - Microemulsion electrokinetic chromatography (MEEKC) was applied to the separation of six catechins and caffeine, the major constituents of the green tea. The developed methods involved the use of sodium dodecyl sulfate (SDS) as surfactant, n-heptane as organic solvent and an alcohol as cosurfactant. The separations were performed under acidic conditions (pH 2.5 phosphate buffer, 50 mM) to ensure good stability of the catechins, with reversed polarity (anodic outlet). The effect of the alcohol nature on the MEEKC selectivity was evaluated; nine alcohols were used as cosurfactant: 1-butanol, tert-butanol, 1-pentanol, 2-pentanol, 3 pentanol, cyclopentanol, 1-hexanol, 2-hexanol, and cyclohexanol. The migration order of (+)-catechin (C), (-)-epicatechin (EC), (-)-epigallocatechin (EGC), (-) gallocatechin (GC), (-)-epigallocatechin gallate (EGCG), (-)-epicatechin gallate (ECG), caffeine and theophylline was significantly affected by the alcohol used as cosurfactant. Using nine microemulsions, four different selectivities were achieved: A (cyclohexanol); B (2-pentanol, 3-pentanol, 1-hexanol, 2-hexanol); C (1-butanol, 1-pentanol, cyclopentanol); D (tert-butanol). MEEKC methods, based on 2-hexanol and cyclohexanol as cosurfactant were validated and successfully applied to the analysis of catechins and caffeine in commercial green tea products. PMID- 12761799 TI - Capillary electrophoresis of cardiolipin with on-line dye interaction and spectrophotometric detection. AB - Cardiolipin is an important phospholipid present in the mitochondrial inner membrane. It plays a key function in mitochondrial respiration by interacting with many enzymes or cofactors related to oxidative phosphorylation complexes. We have determined the concentration of cardiolipin using on-line 10-N-nonyl acridine orange (NAO) dye interaction capillary electrophoresis (CE) and spectrophotometric detection with a sample throughput of 3 min. In addition to the presence of 0.1 mM NAO, the background electrolyte (BGE) composition has been set at 80% methanol-10% acetonitrile-10% H(2)O (all v/v) to provide both good solubility and the maximum absorbance enhancement at 497 nm for the NAO cardiolipin complex as compared to NAO alone. Sample consumption for each injection is about 57 nL. A calibration curve is established from 0.5 microM to 0.1 mM with R (2) = 0.9912 with a detection limit of 0.05 microM for cardiolipin. In a blind study, actual mitochondrial cell membrane samples in the microL range before or after UV light exposure were analyzed using the CE method. Cardiolipin concentration decreased in the different parts of the membrane sample upon UV photolysis of the cells. Support for the theory that UV light can induce cardiolipin translocation from the inner membrane (IM) to the outer membrane (OM) was indicated by a significant percentage increase of cardiolipin (as measured by the cardiolipin in the OM as compared to the sum total in the OM and IM) from 30.7 +/- 2.4% before UV light photolysis to 38.3 +/- 2.2% after UV irradiation. PMID- 12761800 TI - Effect of diluting agent on sensitivity in capillary electrophoresis with amperometric detection. AB - Besides the running buffer, pH of buffer, separation voltage and sampling time, the diluting agent was studied in this paper as one of the important factors influencing the sensitivity in capillary electrophoresis (CE) with amperometric detection (AD) when electrokinectic sampling was used. Clonidine hydrochloride, hydrochlorothiazide and rutin, which are positively charged, neutral and negatively charged, respectively, in aqueous solutions, could be perfectly separated by CE with 25 mmol x L(-1) Na(2)B(4)O(7) - 50 mmol x L(-1) NaH(2)PO(4) as running buffer and detected by measuring their current responses with AD. Before CE running, some kinds of diluents including water, methanol, formamide, running buffer, hydrochloric acid and sodium hydroxide were, respectively, applied to dilute the stock solutions of above three analytes and their effects on the sensitivity of CE-AD were investigated. The results showed that for electrokinetic injection, the current responses of these three analytes were greatly affected in different ways when different diluting agents were used. This method was applied to simultaneously determine the active ingredients in one Chinese compound hypotensor named Zhen Ju Jiang Ya Pian, in which the contents of clonidine hydrochloride, hydrochlorothiazide and rutin is very different as 0.03 mg : 5 mg : 20 mg per tablet, and satisfactory results were obtained by adjusting their sensitivity by selecting the suitable diluting agent. PMID- 12761798 TI - Enantioselective strong cation-exchange molecular recognition materials: design of novel chiral stationary phases and their application for enantioseparation of chiral bases by nonaqueous capillary electrochromatography. AB - New chiral stationary phases derived from enantiomerically pure derivatives of cysteine carrying sulfonic acid groups are synthesized and evaluated for enantiomer separation of chiral bases by non aqueous capillary electrochromatography after bonding to a linker and grafting upon thiol-modified silica particles. Structural modifications of these low molecular weight chiral selectors are investigated and discussed in terms of apparent enantioselectivities and resolution factors based on the enantiomeric separations of a set of chiral bases including beta-blockers, beta-sympathomimetics and other basic drugs. The influence of the mobile phase constitution and its flow velocity on the enantioseparation by nonaqueous capillary electrochromatography is also briefly evaluated and discussed for the chiral substances investigated. PMID- 12761802 TI - Ultrasound-enhanced transdermal transport. AB - The enhancement of transdermal transport by ultrasound is reviewed. After a brief discussion of the physics of ultrasound and its medical applications, the effects of high- and low-frequency ultrasound on the transport of substances across the skin are examined. The impact of low-frequency sonophoresis appears to be much more important, with significant increases in transport into and from the skin following its application. Although the mechanism of action remains incompletely defined, cavitation and thermal processes are strongly implicated. PMID- 12761803 TI - Skin permeability enhancement by low frequency sonophoresis: lipid extraction and transport pathways. AB - The objective of this study was to shed light on the mechanism(s) by which low frequency ultrasound (20 KHz) enhances the permeability of the skin. The physical effects on the barrier and the transport pathway, in particular, were examined. The amount of lipid removed from the intercellular domains of the stratum corneum following sonophoresis was determined by infrared spectroscopy. Transport of the fluorescent probes nile red and calcein, under the influence of ultrasound, was evaluated by laser-scanning confocal microscopy. The results were compared with the appropriate passive control data and with data obtained from experiments in which the skin was exposed simply to the thermal effects induced by ultrasound treatment. A significant fraction ( approximately 30%) of the intercellular lipids of the stratum corneum, which are principally responsible for skin barrier function, were removed during the application of low-frequency sonophoresis. Although the confocal images from the nile red experiments were not particularly informative, ultrasound clearly and significantly (again, relative to the corresponding controls) facilitated transport of the hydrophilic calcein via discrete permeabilized regions, whereas other areas of the barrier were apparently unaffected. Lipid removal from the stratum corneum is implicated as a factor contributing the observed permeation enhancement effects of low-frequency ultrasound. However, microscopic observations imply that sonophoresis induces localized (aqueous?) permeation pathways at discrete sites. PMID- 12761805 TI - A Bayesian method for predicting 5-fluorouracil pharmacokinetic parameters following short-term infusion in patients with colorectal cancer. AB - The objective of this study was to develop a population pharmacokinetic model and validate it using a Bayesian approach for predicting, a priori and a posteriori, the individual volume of distribution (V(d)) and clearance (Cl) of 5-fluorouracil (5-FU) given as short-term intravenous infusion in weekly and multiple doses. Forty-four patients were divided in group A (5-FU weekly doses) including 27 patients with nonmetastatic colorectal adenocarcinoma treated with 450 mg/m(2) of 5-FU, 1 day per week for 48 doses, plus oral levamisol (50 mg/8 h) for 3 days, every 15 days and group B (5-FU multiple doses) including 17 patients with metastatic colorectal adenocarcinoma, receiving 5-FU (425 mg/m(2)) plus intravenous folinic acid (20 mg/m(2)) over 5 consecutive days, every 4 weeks for six cycles. In both groups 5-FU was administered as a 30-60-min infusion. A total of 176 plasma concentrations were analyzed using a NONMEM program according to a linear one-compartment model. In group A, 5-FU population pharmacokinetic parameters were obtained and the covariables studied were age, gender, weight, ideal body weight, height, body surface area, creatinine clearance, and hepatic function tests. A priori and a posteriori validation of this model was carried out with plasma concentrations obtained in day 1 in group B. In group B, population pharmacokinetic parameters of 5-FU following multiple doses were estimated using scale factors to identify differences in 5-FU V(d) and Cl between days 1 and 4, and the interindividual, interoccasion, and residual variabilities studied. V(d) was 0.266 L/kg of ideal body weight and Cl was 1.21 L/h. kg of total weight following weekly doses. The plasma sample obtained at 10 min gave the best accuracy and precision predictions. When 5-FU was administered in multiple doses, the Cl of the drug in day 4 is reduced by 30.14% compared to day 1. The interoccasion variability was lower than interindividual variability for both V(d) and Cl, suggesting that it could be feasible to individualise dosage of 5-FU for subsequent cycles from data obtained in a previous one in an attempt to improve the therapeutic index of colorectal cancer treatment. PMID- 12761804 TI - Hemoperfused isolated porcine slaughterhouse kidneys as a valid model for pharmacological studies. AB - Mammalian models of isolated perfused kidneys provide an important tool to study pharmacological, toxicological, and physiological properties of drugs, hormones, and vasoactive substances. As organs from small laboratory animals are difficult to compare to human conditions, porcine and bovine kidneys permit better approaches to simulate human conditions. We developed an alternative model for pharmacological studies using isolated hemoperfused porcine kidneys from slaughterhouse animals to reduce laboratory animal experiments. Controlled pharmacological studies were established using furosemide (2 mg/100 g organweight) as a model drug. Kidneys were hemoperfused after a preservation period of 4.6 +/- 1.7 h. In comparison to the control period, furosemide application led to significant changes in renal parameters with urine flow: 4.2/1.7 mL/min*100 g (furosemide/control), urine-sodium: 108/77.5 mmol/L, sodium excretion: 0.47/0.14 mmol/min*100 g; all differences significant, p < 0.01. The parameters stabilized to normal values as found in the control period within a period of 80 min. A second group of laboratory-harvested kidneys was examined for differences and revealed limitations of the slaughterhouse organs in parameters such as oxygen consumption. In summary, the present study demonstrates the valid use of hemoperfused slaughterhouse kidneys as a pharmacological model of renal function within the limits of the use of slaughterhouse organs, and indicates that future studies using this alternative approach could reduce animal experiments. PMID- 12761806 TI - Once-a-day controlled-release dosage form of divalproex sodium I: formulation design and in vitro/in vivo investigations. AB - Divalproex sodium is a narrow therapeutic index drug that is widely used for the treatment of epilepsy, the manic episodes associated with bipolar disorder, and prophylaxis of migraine headaches. The present investigation was undertaken to design an oral dosage form that would provide once-daily administration with improved therapy and to explore the relationships between in vitro drug release and in vivo absorption. Controlled release hydrophilic matrix formulations of divalproex sodium were designed and evaluated via in vitro and in vivo studies. The release rate of divalproex sodium was modulated by varying different rate controlling hydrophilic polymers and measured in vitro using a USP apparatus II dissolution method. Formulations with differing release rates were studied in beagle dogs and in healthy subjects. A selected formulation given once-daily was further evaluated against the commercial enteric tablet dosed twice-daily in a multiple dose study, and shown to provide desired nearly constant therapeutic plasma concentrations over the entire 24-h dosing interval. Preliminary linear relationships between in vitro dissolution and in vivo absorption were observed in both the animal model and in humans. However, the relationships were formulation dependent, indicating a need for further studies. PMID- 12761807 TI - In vivo gene transfer using sulfhydryl cross-linked PEG-peptide/glycopeptide DNA co-condensates. AB - Recent interest in sulfhydryl cross-linked nonviral gene delivery systems, designed to trigger the intracellular release of DNA, has inspired studies to establish their utility in vitro. To determine if this concept can be extrapolated to in vivo gene delivery, sulfhydryl cross-linking peptides (dp 20), derivatized with either an N-glycan or polyethylene glycol (PEG), were used to generate sulfhydryl cross-linked gene formulations. The biodistribution, metabolism, cell-type targeting, and gene expression of sulfhydryl cross-linked PEG-peptide/glycopeptide DNA co-condensates were examined following i.v. dosing in mice. Optimal targeting to hepatocytes was achieved by condensing (125)I-DNA with an add-mixture of 10 mol % triantennary glycopeptide, 5 mol % PEG-peptide, and 85 mol % backbone peptide. Four backbone peptides were substituted into the formulation to examine the influence of peptide metabolism and disulfide bond strength on the rate of DNA metabolism and the level of gene expression in vivo. The half-life of DNA in liver was extended from 1 to 3 h using a backbone peptide composed of d-amino acids, whereas substituting penicillamine for cysteine failed to further increase the metabolic stability of DNA. Optimized gene delivery formulations transiently expressed secreted alkaline phosphatase in mouse serum for 12 days. The results suggest that disulfide bond reduction in liver hepatocytes proceeds rapidly, followed by peptide metabolism, ultimately limiting the metabolic half-life of sulfhydryl cross-linked DNA condensates in vivo. PMID- 12761808 TI - A method to protect sensitive molecules from a light-induced polymerizing environment. AB - Systems that can be polymerized in situ upon exposure to light radiation may have significant applications in tissue engineering and drug delivery. However, the light-induced polymerization step, which is the requisite for this technology, could be potentially deleterious to sensitive bioactive agents (e.g., enzymes, cytokines, matrix metalloproteinases) being entrapped. In this study, a method to protect sensitive molecules from a light-induced polymerizing environment is proposed. This method is based on the idea that nonaccessible substances cannot interact with the polymerizing species. To examine this concept, two model enzymes-namely, horseradish peroxidase and alpha-glucosidase-were protected by gelatin-based wet granulation and incorporated within a cured polyethylene glycol dimethacrylate, a photocurable monomer, under different conditions. Unprotected enzymes were used as controls. Enzymes were then allowed to diffuse out of the polymerized matrices. The activity and total enzyme recovered from these matrices by passive diffusion were compared to ascertain the extent of activity retention. Matrix assisted laser desorption ionization mass spectrometry combined with time of flight mass spectrometry (MALDI-TOF) was used to determine changes in enzyme molecular weight. During the first 24 h of diffusion from the polymerized matrices, unprotected enzymes consistently showed a loss of activity ranging from 10-66%, depending on the matrix composition and enzyme properties. In contrast, protected enzymes retained over 94% of their activity irrespective of the experimental setting. The loss of activity appears to be a direct consequence of the polymerizing environment. PMID- 12761809 TI - Crystal forms of LY334370 HCl: isolation, solid-state characterization, and physicochemical properties. AB - LY334370 HCl, a 5HT1f agonist investigated for the treatment of migraines, was identified in five crystal forms: three anhydrates (I-III), a dihydrate, and an acetic acid solvate. The identification and characterization of these crystal forms by optical microscopy, differential scanning calorimetry, thermogravimetric and moisture sorption analyses, solid-state NMR spectroscopy, and X-ray crystallography (Form I only) are presented. Physical properties, including hygroscopicity, solubility, and intrinsic dissolution rate, were assessed for Form I and the dihydrate, the two most viable crystal forms for commercial development. Surprisingly, anhydrous Form I was found to be the thermodynamically most stable crystal form in water, dissolving six times slower than the dihydrate, a difference that correlates well with the rank order of aqueous solubility. PMID- 12761810 TI - Pharmacokinetic/pharmacodynamic modeling of the effects of intravenous immunoglobulin on the disposition of antiplatelet antibodies in a rat model of immune thrombocytopenia. AB - Recently, our laboratory reported that intravenous immunoglobulin (IVIG) treatment increased antiplatelet antibody (7E3) clearance in a rat model of immune thrombocytopenic purpura (ITP). However, due to the multifaceted nature of IVIG therapy, the contribution of this increase in antiplatelet antibody clearance to the total therapeutic effect of IVIG was unclear. The purposes of the present study were to (1). develop a new, mechanistic model of immune gamma globulin (IgG) pharmacokinetics, (2). develop a pharmacokinetic/pharmacodynamic (PK/PD) model relating 7E3 concentrations to the platelet count time course observed following 7E3 treatment, and (3). use these mathematical models to gain insight into the significance of increased 7E3 clearance relative to the total effect of IVIG on 7E3-induced thrombocytopenia. A mechanism-based PK model was developed that adequately characterized IVIG effects on 7E3 pharmacokinetics. The structure of this model is based on competition between IgG molecules for occupancy of the protective FcRn receptor. The model accurately captured antiplatelet antibody concentration versus time data in the presence and absence of IVIG therapy, in the rat, in 'wild-type' mice, and in 'knockout' mice lacking expression of the FcRn receptor. An indirect response PK/PD model was also developed, which accurately characterized 7E3 effects on platelet counts. Using these models, it was estimated that 50 +/- 11% of the total protective effect of IVIG in this acute model of ITP can be accounted for by IVIG effects on 7E3 disposition. PMID- 12761811 TI - Pulmonary absorption rate and bioavailability of drugs in vivo in rats: structure absorption relationships and physicochemical profiling of inhaled drugs. AB - The aim of this investigation was to analyze the structure-absorption relationships for pulmonary delivered drugs. First, the inhaled drugs on the market during 2001 were identified and a profile of the calculated physicochemical properties was made. Second, an in vivo pharmacokinetic investigation was performed in anesthetized rats. Eight selected drugs were administered by intratracheal nebulization and intravenous bolus administration and the plasma concentrations of the drugs were determined by LC-MS-MS. Third, an evaluation of the relationships between the absorption/bioavailability data and the drugs' physicochemical characteristics and the epithelial permeability in Caco-2 cells, respectively, was performed. The drug absorption rate was found to correlate to the molecular polar surface area and the hydrogen bonding potential, as well as to the apparent permeability in Caco-2 cell monolayers, which indicated that passive diffusion was the predominating mechanism of absorption in the rat lung. In contrast to the intestinal mucosa and the blood-brain barrier, the pulmonary epithelium was shown to be highly permeable to compounds with high molecular polar surface area (e.g., PSA 479 A(2)). Furthermore, a high bioavailability was found for the efflux transporter substrates talinolol (81%) and losartan (92%), which provides functional evidence for a quantitatively less important role for efflux transporters, such as P-glycoprotein, in limiting the absorption of these drugs from the rat lung. In conclusion, the pulmonary route should be regarded as a potential alternative for the delivery of drugs that are inadequately absorbed after oral administration. PMID- 12761812 TI - Improvement in the in situ gelling properties of deacetylated gellan gum by the immobilization of thiol groups. AB - The rheological properties of an in situ crosslinking thiolated deacetylated gellan gum were examined in vitro. Mediated by a carbodiimide, L-cysteine was covalently bound to deacetylated gellan gum (DGG). The deacetylated gellan gum cysteine (DGG-Cys) conjugate displayed 216.53 +/- 59.54 micromol thiol groups per gram polymer (means +/- SD, n = 3). The thiolated polymer was capable of forming inter- and/or intramolecular disulfide bonds in aqueous solution (1.5%; m/m) at pH 7. After 6 h of incubation at room temperature, storage modulus, loss modulus, and complex viscosity increased 300-, 6.4-, and 26.6-fold, respectively, relative to the unthiolated polymer. Loss tangent of DGG-Cys was <1, indicating a gel, whereas the corresponding unmodified polymer had a loss tangent of >1, indicating a fluid. Frequency sweep measurements demonstrated an increase in crosslinking of the thiolated polymer as a function of time. DGG-Cys appeared to be superior to the unmodified polymer also in the presence of physiological cation concentrations found (e.g., in tear fluid and nasal secretion), which is referred to rheological properties. The polymer generated within this study represents a promising novel excipient for various drug delivery systems in which in situ gelling properties are favorable. PMID- 12761813 TI - Simultaneous measurement of drug release and liquid uptake in pharmaceutical tablets. AB - A new method is introduced that allows drug release and liquid absorption to be studied simultaneously. The drug release is measured by the alternating ionic current (AIC) method, and the study of liquid uptake is accomplished with a sensitive microbalance from a processor tensiometer. We show that the method can be employed to study anomalous diffusion in the initial phase of the drug release process of disintegrating systems. We also demonstrate that the diffusion layer thickness and the diffusion coefficient in a dissolving system can be obtained with the new measurement technique. PMID- 12761814 TI - The role of surfactants in the reversal of active transport mediated by multidrug resistance proteins. AB - A variety of seven nonionic, one amphoteric and, one anionic surfactant that are applied or investigated as surfactants in drug formulation, were analyzed for their capacity to modulate carrier-mediated transport by efflux pumps. Two cell lines, murine monocytic leukemia cells overexpressing P-glycoprotein (P-gp) and Madin-Darby canine kidney cells stably overexpresssing human multidrug resistance associated protein 2 (MRP2), were used as test systems. The modulation of P-gp and of MRP2 function was studied by the reversal of rhodamine 123 and of methylfluorescein-glutathione conjugate transport, respectively. Mechanisms that were not transporter related and could lead to misinterpretations were identified, such as probe quenching, probe encapsulation by micelles, and membrane damage. P-gp-mediated rhodamine 123 transport was inhibited by five nonionic surfactants in a concentration-dependent manner and in the order TPGS > Pluronic PE8100 > Cremophor EL > Pluronic PE6100 approximately Tween 80. In contrast, none of the surfactants showed a significant inhibition of MRP2 mediated efflux in Madin-Darby canine kidney/MRP2 cells. In conclusion, the results indicate that surfactants demonstrate a transporter-specific interaction, rather than unspecific membrane permeabilization. The present analysis offers insight in the possible mechanisms of surfactant interactions with biological membranes and could help to identify specific drug formulations. PMID- 12761815 TI - Using the polymer partitioning method to probe the thermodynamic activity of poorly water-soluble drugs solubilized in model lipid digestion products. AB - The thermodynamic activity of solubilized drug is an important determinant of the extent of absorption of lipophilic drugs from the gastrointestinal tract. In this study, the polymer partitioning method was evaluated for its use in the determination of the thermodynamic activity of lipophilic drugs when solubilized in colloidal digestion products, using drug in dilute solution as a reference ideal solution. The lipophilic drugs griseofulvin, diazepam, and danazol partitioned into a polymeric receiver phase from non-micellar solution as a function of drug lipophilicity. The concentration of drug that partitioned into the polymer was linearly proportional to the concentration of free drug in solution, and this allowed the measured partition coefficient to be utilized as an indicator of the drug activity coefficient. The addition of a solubilizing species such as bile salt micelles caused a reduction in drug activity of a similar magnitude to that predicted from micelle equilibrium solubility data in the identical micellar solutions. The addition of micelle swelling lipids such as lecithin and fatty acids resulted in further reductions in activity coefficient. The ability to measure drug activity in model digestive systems has potential for application in the rational development of improved lipid-based formulations of poorly water-soluble drugs for oral administration. PMID- 12761816 TI - A fluorescence study of the structure and accessibility of plasmid DNA condensed with cationic gene delivery vehicles. AB - The cationic lipids 1,2-dioleoyl-3-trimethylammonium-propane and dimethyldioctadecylammonium bromide, with or without the helper lipids 1,2 dioleoyl-sn-glycero-3-phosphoethanolamine or cholesterol, and the cationic polymer polyethyleneimine, were compared for their ability to displace fluorescent dyes from DNA. Differences in displacement of the intercalating dyes ethidium bromide and ethidium homodimer correlate with their relative affinities with DNA, with the extent of ethidium homodimer displacement significantly less. Differences in ethidium homodimer and ethidium bromide displacement as a function of the ratio of polycation to DNA and the charge density of the polycation suggest a greater sensitivity of the former to topological changes in condensed DNA. Marked differences in the ability of these cationic delivery systems to displace the minor groove binding dyes 4',6-diamidino-2-phenylindole and Hoechst 33258 upon interaction with DNA are also apparent, with the majority of Hoechst 33258 remaining bound to DNA. Changes in the spectral properties of Hoechst 33258 were further used to characterize polycation-induced changes in solvent accessibility of the DNA minor groove. Taken together, these studies demonstrate differences in the interaction of various cationic lipids and polyethyleneimine in terms of regional displacement of dyes, polycation-induced structural changes in DNA, as well as polycation-mediated changes in solvent accessibility of the minor groove. The relevance of these studies to current models of the structure and assembly of polycation/DNA complexes are discussed. PMID- 12761817 TI - Inverse gas chromatography: considerations about appropriate use for amorphous and crystalline powders. AB - The use of inverse gas chromatography to assess surface properties of a range of pharmaceutical powders was examined. The powders were two sources of hydroxy propylmethyl cellulose (HPMC), microcrystalline cellulose, magnesium stearate, and acyclovir. These were selected to cover a range for properties from amorphous to crystalline, hydrophilic to hydrophobic, and high to low aqueous solubility. It was found that many powders gave a similar value for the dispersive surface energy, which is surprising given the differences in chemical nature. It is likely that this is due to the use of infinite dilution giving rise to the study of specific regions of the powder surface only. The values obtained for dispersive energies were not influenced by packing mass or flow rate of the carrier gas. The retention of polar probes on the column was a concern for the amorphous HPMC samples. This gave rise to derived values for acid-base nature which varied depending on sample mass and carrier gas flow rate. The data show that care must be taken when studying amorphous samples for which it is possible to obtain diffusion into the material rather than just surface adsorption of probes. Despite these problems, it was still possible to differentiate between the samples (including differences between the two HPMC samples) by use of polar probes. It was also possible to see differences in absorption into the sample, reflecting the different physical forms. For example, microcrystalline cellulose behaved very differently to HPMC. It can be concluded that inverse gas chromatography is a valuable characterization tool, but it must be used with care especially with respect to polar probes on amorphous samples. PMID- 12761818 TI - Disposition and gene expression characteristics in solid tumors and skeletal muscle after direct injection of naked plasmid DNA in mice. AB - Previous studies have suggested that direct injection of naked plasmid DNA (pDNA) into solid tumors can be a useful method for in vivo gene transfer into tumor cells. To gain more insight into this approach, we studied the disposition and gene expression characteristics of naked pDNA after intratumoral injection by direct comparison with those after intramuscular injection in mice. pDNA encoding reporter genes were directly injected into subcutaneous solid tumor models and skeletal muscles. Biodistribution studies using radiolabeled pDNA showed that the elimination of pDNA from the injection site was relatively fast and a part of the pDNA was absorbed from the lymphatic system after both local injections. Confocal microscopic studies using fluorescein-labeled pDNA demonstrated that pDNA distributed efficiently throughout the muscle tissue whereas pDNA localization in the tumor tissue was restricted. Characterization of gene expression clarified the variation in expression level between tumor preparations and some factors affecting the expression level in the tumor. Reporter gene expression was significantly inhibited by simultaneous administration of some polyanions in both cases, suggesting that a specific mechanism may be involved in the naked pDNA uptake by muscle and tumor cells. These findings provide useful information for direct naked pDNA delivery into solid tumors. PMID- 12761819 TI - Structure-activity relationship for chemical skin permeation enhancers: probing the chemical microenvironment of the site of action. AB - Studies were previously conducted in our laboratory on the influence of n alkanols, 1-alkyl-2-pyrrolidones, N,N-dimethlyalkanamides, and 1,2-alkanediols as skin permeation enhancers on the transport of a model permeant, corticosterone (CS). The experiments were conducted with hairless mouse skin (HMS) in a side-by side, two-chamber diffusion cell, with enhancer present in an aqueous buffer in both chambers. The purpose of the present study was to extend these studies and investigate in greater detail the hypothesis that a suitable semipolar organic phase may mimic the microenvironment of the site of enhancer action, and that the enhancer partitioning tendency into this organic phase may be used to predict the enhancer potency. CS flux enhancement along the lipoidal pathway of HMS stratum corneum was determined with the 1-alkyl-2-azacycloheptanones, 1-alkyl-2 piperidinones, 1,2-dihydroxypropyl decanoate, 1,2-dihydroxypropyl octanoate, n alkyl-beta-D-glucopyranosides, 2-(1-alkyl)-2-methyl-1,3-dioxolanes, 1,2,3 nonanetriol, and trans-hydroxyproline-N-decanamide-C-ethylamide as enhancers. Enhancement factors (E values) were calculated from the permeability coefficient and solubility data over a range of E values. Comparisons of the enhancer potencies for all studied homologous series and the carbon number of the n-alkyl group revealed a nearly semilogarithmic linear relationship with a slope of approximately 0.55, which is consistent with the hydrophobic effect. Moreover, comparisons of the enhancer potencies of all the enhancers with the n-hexanol phosphate buffered saline (PBS), n-octanol-PBS, n-decanol-PBS, and n-hexane-PBS partition coefficients showed very good correlations for the n-alkanol solvents but not for n-hexane. This result supports the interpretation that the enhancer potency is directly related to the ability of the enhancer molecule to translocate to a site of action via its free energy of transfer from the bulk aqueous phase to a semipolar microenvironment in the stratum corneum lipid lamella that is well mimicked by water-saturated n-alkanols. PMID- 12761821 TI - Glutamate receptor subunit 3 (GluR3) immunoreactivity delineates a subpopulation of parvalbumin-containing interneurons in the rat hippocampus. AB - Rasmussen's encephalitis is a childhood disease resulting in intractable seizures associated with hippocampal and neocortical inflammation. An autoantibody against the GluR3 subunit of alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA) receptors is implicated in the pathophysiology of Rasmussen's encephalitis. AMPA receptors mediate excitatory neurotransmission in the brain and contain combinations of four subunits (GluR1-4). Although the distributions of GluR1, GluR2, and GluR4 are known in some detail, the cellular distribution of GluR3 in the mammalian brain remains to be described. We developed and characterized a GluR3-specific monoclonal antibody and quantified the cellular distribution of GluR3 in CA1 of the rat hippocampus. GluR3 immunoreactivity was detected in all pyramidal neurons and astrocytes and in most interneurons. We quantified the intensity of GluR3 immunoreactivity in interneuron subtypes defined by their calcium-binding protein content. GluR3 immunofluorescence, but not GluR1 or GluR2 immunofluorescence, was significantly elevated in somata of parvalbumin-containing interneurons compared to pyramidal somata. Strikingly, increased GluR3 immunofluorescence was not observed in calbindin- and calretinin containing interneurons. Furthermore, 24% of parvalbumin-containing interneurons could be distinguished from surrounding neurons based on their intense GluR3 immunoreactivity. This subpopulation had significantly elevated GluR3 immunoreactivity compared to the rest of parvalbumin-containing interneurons. Electron microscopy revealed enriched GluR3 immunoreactivity in parvalbumin containing perikarya at cytoplasmic and postsynaptic sites. Parvalbumin containing interneurons, potent inhibitors of cortical pyramidal neurons, are vulnerable in the brains of epileptic patients. Our findings suggest that the somata of these interneurons are enriched in GluR3, which may render them vulnerable to pathological states such as epilepsy and Rasmussen's encephalitis. PMID- 12761822 TI - Dermatomes and the central organization of dermatomes and body surface regions in the spinal cord dorsal horn in rats. AB - Dermatomes and the associated central projection fields were studied with the application of fluorescent neurotracer, 1,1'-dioctadecyl-3,3,3',3' tetramethylindocarbocyanine perchlorate (DiI), to 21 reference points on rat trunk and hindlimb skin. Segmental distribution and rostrocaudal central level of dorsal root ganglion (DRG) neurons innervating reference points were examined and DiI-induced fluorescent areas were mapped in the horizontal plane through lamina II of the dorsal horn. Segmental levels of DRG neurons innervating reference points were generally identical to the level determined using dye-extravasation methods. However, innervation of the first digit was situated in the L4 dermatome, not the L3 reported previously using those methods. Generally, afferents from a reference point projected to a single field in the ipsilateral dorsal horn. Reference points on ventral and dorsal median lines of the trunk were represented bilaterally. Afferents from reference points located on the ventral median line of the hindlimb projected to two separate fields: one on the medial margin of spinal cord segments L2-L5 and the other on the medial half of spinal cord segment L5. From the distribution of central projection fields of reference points, central projection fields of dermatomes were revealed as even in shape and located within corresponding spinal cord segments. The arrangement of peripheral and central fields of dermatomes and body surface regions suggests that peripheral and central projection fields of cutaneous afferent fibers are reshaped from the common prototypical pattern that exhibits an orderly and evenly sequenced arrangement. PMID- 12761820 TI - Putative isotocin distributions in sonic fish: relation to vasotocin and vocal acoustic circuitry. AB - Recent neurophysiological evidence in the plainfin midshipman fish (Porichthys notatus) demonstrated that isotocin (IT) and arginine vasotocin (AVT) modulate fictive vocalizations divergently between three reproductive morphs. To provide an anatomical framework for the modulation of vocalization by IT and to foster comparisons with the distributions of the IT homologues mesotocin (MT) and oxytocin (OT) in other vertebrate groups, we describe putative IT distributions in the midshipman and the closely related gulf toadfish, Opsanus beta. Double label fluorescent histochemistry was used for IT and AVT (by using antibodies for MT, OT, and the mammalian AVT homologue, arginine vasopressin [AVP]). MT/OT-like immunoreactive (MT/OT-lir) cell groups were found in the anterior parvocellular, posterior parvocellular, and magnocellular preoptic nuclei. MT/OT-lir fibers and putative terminals densely innervated the ventral telencephalon and numerous areas in the hypothalamus and brainstem. These distributions included all sites of vocal-acoustic integration recently identified for the forebrain and midbrain and diencephalic components of the ascending auditory pathway. Results were qualitatively comparable across morphs, species, and seasons. In contrast to the widespread distribution of MT/OT-lir, AVP-lir somata, fibers, and putative terminals were almost completely restricted to vocal-acoustic regions. These data parallel previous descriptions of AVT immunoreactivity in these species, although the present methods showed a previously undescribed, seasonally variable AVP-lir cell group in the anterior tuberal hypothalamus, a vocally active site and a component of the ascending auditory pathway. These findings provided anatomic support for the role of IT and AVT in the modulation of vocal behavior at multiple levels of the central vocal-acoustic circuitry. PMID- 12761823 TI - VEGF expression by ganglion cells in central retina before formation of the foveal depression in monkey retina: evidence of developmental hypoxia. AB - In macaque monkeys the foveal depression forms between fetal day (Fd) 105 and birth (Fd 172 of gestation). Before this, the incipient fovea is identified by a photoreceptor layer comprising cones almost exclusively, a multilayered ganglion cell layer (GCL), and a "domed" profile. Vessels are absent from the central retina until late in development, leading to the suggestion that the GCL in the incipient fovea may be transitorily hypoxic. Vascular endothelial growth factor (VEGF), expressed by both glial and neuronal cells and mediated by the hypoxia inducible transcription factor (HIF)-1, is the principal factor involved in blood vessel growth in the retina. We examined VEGF expression in macaque retinas between Fd 85 and 4 months postnatal. Digoxygenin-labeled riboprobes were generated from a partial-length human cDNA polymerase chain reaction fragment, detected using fluorescence confocal microscopy, and quantified using Scion Image. High levels of VEGF mRNA were detected in astrocytes associated with developing vessels. We also detected strong expression of VEGF mRNA in the GCL at the incipient fovea prior to Fd 105, with peak labeling in the incipient fovea that declined with distance in nasal and temporal directions. By Fd 152 peak labeling was in two bands associated with development of the inner nuclear layer (INL) capillary plexus: in the inner INL where Muller and amacrine cell somas are located, and in the outer INL where horizontal cells are found. The findings suggest that at the incipient fovea the GCL is hypoxic, supporting the hypothesis that the adaptive significance of the fovea centralis is in ensuring adequate oxygen supply to neuronal elements initially located within the avascular region. PMID- 12761824 TI - Octopaminergic modulation of synaptic transmission between an identified sensory afferent and flight motoneuron in the locust. AB - The role of the biogenic amine octopamine in modulating cholinergic synaptic transmission between the locust forewing stretch receptor neuron (fSR) and the first basalar motoneuron (BA1) was investigated. The amines 5-hydroxytryptamine (5-HT, serotonin) and dopamine were also studied. Bath application of octopamine, 5-HT, and dopamine at concentrations of 10(-4) M reversibly decreased the amplitude of monosynaptic excitatory postsynaptic potentials (EPSPs) evoked in BA1 by electrically stimulating the fSR axon. These effects occurred without any detectable change in either input resistance or membrane potential of BA1. The amines also reversibly decreased the amplitude of responses to acetylcholine (ACh) pressure-applied to the soma of BA1. The muscarinic antagonist scopolamine (10(-6) M) had no significant effect on the octopamine-induced decrease in ACh responses. These observations suggest that these amines potentially could physiologically depress cholinergic transmission between fSR and BA1, at least in part, by altering nicotinic rather than muscarinic cholinergic receptor function. Although the octopaminergic agonists naphazoline and tolazoline both mimicked the actions of octopamine, the receptor responsible for octopamine-mediated modulation could not be characterized since amine receptor antagonists tested on the preparation had complex actions. Confocal immunocytochemistry revealed intense octopamine immunoreactivity in the anterior lateral association center, thus confirming the presence of octopamine in neuropil regions containing fSR/BA1 synapses and therefore supporting a role for this amine in the modulation of synaptic transmission between the fSR and BA1. 5-HT-immunoreactivity, conversely, was concentrated within the ventral association centers; very little staining was observed in the dorsal neuropil regions in which fSR/BA1 synapses are located. PMID- 12761825 TI - Lysosomal amino acid transporter LYAAT-1 in the rat central nervous system: an in situ hybridization and immunohistochemical study. AB - A first mammalian lysosomal transporter (LYAAT-1) was recently identified and functionally characterized. Preliminary immunocytochemical data revealed that LYAAT-1 localizes to lysosomes in some neurons. In order to determine whether it is expressed in specific neuron populations and other cell types, and to confirm whether it is localized at the membrane of lysosomes, we used in situ hybridization and immunohistochemistry methods in adult rat central nervous system (CNS). We found that LYAAT-1 is expressed in most areas of the CNS, specifically in neurons, but also in choroid plexus and ependymal epithelium cells. LYAAT-1-IR (immunoreactivity) levels varied among different neuroanatomical structures but were present in neurons independently of the neurotransmitter used (glutamate, GABA, acetylcholine, noradrenaline, serotonin, or glycine). Light and confocal microscopy demonstrated that LYAAT-1 and the lysosomal marker cathepsin D colocalized throughout the brain and electron microscopy showed that LYAAT-1-IR was associated with lysosomal membranes. In addition, LYAAT-1-IR was also found associated with other membranes belonging to the Golgi apparatus and lateral saccules and less frequently with multivesicular bodies, endoplasmic reticulum, and occasionally with the plasma membrane. The localization of LYAAT-1 at the lysosomal membrane is consistent with the view that it mediates amino acid efflux from lysosomes. Furthermore, its cell expression pattern suggests that it may contribute to specialized cellular function in the rat CNS such as neuronal metabolism, neurotransmission, and control of brain amino acid homeostasis. PMID- 12761826 TI - EphA4 provides repulsive signals to developing cochlear ganglion neurites mediated through ephrin-B2 and -B3. AB - The ephrins and Eph receptors make up two large families of bi-directional signaling molecules that are known to play a role in the development of the nervous system. Recently, expression of EphA4 in the developing cochlea was shown, with strong expression in cells lining the osseous spiral lamina (OSL) through which afferent dendrites must pass to reach the organ of Corti (OC). It was also demonstrated that ephrin-B2 and -B3, both of which are known to interact with EphA4, are expressed by spiral ganglion (SG) neurons. To investigate the functional role of EphA4 in the development of inner ear neurons, neonatal rat SG explants were cultured for 72 hours on uniformly coated surfaces or near stripes of EphA4/IgG-Fc-chimera. Control explants were cultured on or near IgG-Fc and EphA1/IgG-Fc-chimera. To assess the roles of ephrin-B2 and -B3 in EphA4 signaling, SG explants were cultured with or without anti-ephrin-B2 and/or -B3 blocking antibodies. Growth patterns of SG neurites at the border of EphA4 receptor stripes showed repulsion, characterized by turning, stopping and/or reversal. In the case of IgG-Fc and EphA1, the neurites grew straight onto the stripes. Treatment with either anti-ephrin-B2 or -B3 blocking antibodies significantly reduced the repulsive effect of an EphA4 stripe. Moreover, when both antibodies were used together, neurites crossed onto EphA4 stripes with no evidence of repulsion. The results suggest that EphA4 provides repulsive signals to SG neurites in the developing cochlea, and that ephrin-B2 and -B3 together mediate this response. PMID- 12761827 TI - Expression of the Kv1.1 ion channel subunit in the auditory brainstem of the big brown bat, Eptesicus fuscus. AB - Voltage-gated potassium channels play an important role in shaping membrane properties that underlie neurons' discharge patterns and the ways in which they transform their input. In the auditory system, low threshold potassium currents such as those created by Kv1.1 subunits contribute to precise phaselocking and to transient onset responses that provide time markers for temporal features of sounds. The purpose of the present study was to compare information about the distribution of neurons expressing the KV 1.1 in the brainstem auditory nuclei with the distribution of neurons with known functional properties in the auditory system of the big brown bat, Eptesicus fuscus. We used immunocytochemistry and light microscopy to look at the distribution of Kv1.1 subunits in the brainstem auditory nuclei. There was prominent expression in cell types known to contain high levels of Kv1.1 in other species and known to respond to auditory signals with high temporal precision. These included octopus cells and spherical bushy cells of the cochlear nucleus and principal neurons of the medial nucleus of the trapezoid body. In addition, we found high levels of Kv1.1 in neurons of the columnar subdivision of the ventral nucleus of the lateral lemniscus and in ventral periolivary cell groups. Neurons with high levels of Kv1.1 were differentially distributed in the intermediate nucleus of the lateral lemniscus and in the inferior colliculus, suggesting that these structures contain functionally distinct cell populations, some of which may be involved in high precision temporal processing. PMID- 12761828 TI - Thalamocortical and intracortical connections of monkey cingulate motor areas. AB - Although there has been an increasing interest in motor functions of the cingulate motor areas, data concerning their input organization are still limited. To address this issue, the patterns of thalamic and cortical inputs to the rostral (CMAr), dorsal (CMAd), and ventral (CMAv) cingulate motor areas were investigated in the macaque monkey. Tracer injections were made into identified forelimb representations of these areas, and the distributions of retrogradely labeled neurons were analyzed in the thalamus and the frontal cortex. The cells of origin of thalamocortical projections to the CMAr were located mainly in the parvicellular division of the ventroanterior nucleus and the oral division of the ventrolateral nucleus (VLo). On the other hand, the thalamocortical neurons to the CMAd/CMAv were distributed predominantly in the VLo and the oral division of the ventroposterolateral nucleus-the caudal division of the ventrolateral nucleus. Additionally, many neurons in the intralaminar nuclear group were seen to project to the cingulate motor areas. Except for their well-developed interconnections, the corticocortical projections to the CMAr and CMAd/CMAv were also distinctively preferential. Major inputs to the CMAr arose from the presupplementary motor area and the dorsal premotor cortex, whereas inputs to the CMAd/CMAv originated not only from these areas but also from the supplementary motor area and the primary motor cortex. The present results indicate that the CMAr and the caudal cingulate motor area (involving both the CMAd and the CMAv) are characterized by distinct patterns of thalamocortical and intracortical connections, reflecting their functional differences. PMID- 12761829 TI - Thromboresistance of alkali- and heat-treated titanium metal formed with apatite. AB - Fibrin deposition and platelet adhesion onto alkali- and heat-treated titanium metal (AH-Ti), alkali- and water-treated titanium metal (Wa-Ti), and alkali- and heat-treated titanium metal formed with apatite (Ap-Ti) in simulated body fluid (SBF) were evaluated by exposure to anticoagulated blood or washed platelet suspension (WPS) under static conditions and subsequent observation with scanning electron microscopy (SEM). The results were compared with those for commercially pure titanium metal (cp-Ti). Thrombus formation on AH-Ti and Wa-Ti, which were exposed to heparinized whole blood for 1 h, was significantly less than that on cp-Ti, on which pronounced depositions of fibrin-erythrocytes and lymphocytes were observed. No thrombus was observed on Ap-Ti, possibly because of a high adsorption of heparin. Morphological change of platelets attached to surfaces via adsorbed plasma proteins was found to a significant extent on AH-Ti and Wa-Ti exposed to WPS. However, there was almost no difference between cp-Ti and Ap-Ti in round morphology of adherent platelets. These findings suggested that Ap-Ti exhibits stronger antithrombogenic characteristics than cp-Ti and other materials examined in heparinized blood. PMID- 12761830 TI - Gas plasma etching of PEO/PBT segmented block copolymer films. AB - A series of poly(ethylene oxide)/poly(butylene terephthalate) (PEO/PBT) segmented block copolymer films was treated with a radio-frequency carbon dioxide (CO(2)) or with argon (Ar) plasma. The effects of (preferential) etching on surface structure, topography, chemistry, and wettability were studied by scanning electron microscopy, atomic force microscopy, X-ray photoelectron spectroscopy, and contact angle measurements. In all cases, a granular-type nanostructure was formed after prolonged CO(2) plasma etching. Ar plasma etching generally did not lead to significant changes in surface structure. Regarding surface chemistry, CO(2) plasma treatment caused surface oxidation and oxidative degradation of the films while Ar plasma etching resulted mainly in the preferential removal of PEO blocks. The wettability of all films significantly increased after plasma treatment because of the creation of polar functional groups at the surface. Preliminary goat bone-marrow cell compatibility experiments have shown that all plasma-treated PEO/PBT films induced a greatly enhanced cell adhesion and/or growth compared to untreated biomaterials. This improvement was attributed to changes in surface chemistry during plasma etching rather than to changes in surface structure. These results show that plasma-treated PEO/PBT copolymers have a high potential as scaffolds for bone tissue regeneration. PMID- 12761831 TI - Adhesion of human leukocytes to biomaterials: an in vitro study using alkanethiolate monolayers with different chemically functionalized surfaces. AB - The adhesion of human leukocytes to self-assembled monolayers of well-defined surface chemistry was investigated in vitro. Polymorphonuclear (PMN) and mononuclear leukocytes were isolated from human blood by centrifugation techniques. The effect on adhesion of cell activation produced by pre-incubation of leukocytes with phytohemagglutinin (PHA) and phorbol 12-myristate 13-acetate (PMA) was also studied. Gold substrates were modified by treatment with alkanethiols with three different terminal chemical groups: COOH, OH, and CH(3). After incubation with the two subpopulations of leukocytes, the monolayers were washed, treated with fixative, stained with a Giemsa method, and observed by light microscopy to quantify the number of attached leukocytes. Comparative quantification of the density of leukocyte adhesion to the three types of self assembled monolayers was determined. The hydrophobic surface expressing CH(3) was found to be the one that induced the highest adhesion density of leukocytes, both of PMN and mononuclear cells. In vitro activation of both mononuclear and PMN leukocytes further increased cell adhesion to the chemically defined monolayers that were used. This enhancement was higher for PHA-activated than for PMA stimulated mononuclear cells, whereas PMA treatment of neutrophils resulted in a higher rate of adhesion of these cells than PHA stimulation. PMID- 12761832 TI - Fibrin as a matrix for grafting 2-hydroxyethyl methacrylate: preparation and characterization of the graft and its in vivo evaluation for wound healing. AB - In this work, fibrin was used as a substrate to graft 2-hydroxyethyl methacrylate (HEMA) by free radical polymerization using potassium persulfate and sodium metabisulfite as redox initiators. The extent of grafting the synthetic polymer on the biopolymer was studied under various experimental conditions, and the optimum factors for affording maximum grafting were standardized. The graft, fib g-p[HEMA], was characterized by Fourier transform infrared, scanning electron microscopy, and X-ray diffraction studies. The graft exhibited a higher shelf life than native fibrin. The biocompatibility of the graft has been tested by in vivo studies and the results, in terms of collagen formation and wound size, proved its suitability for wound healing. PMID- 12761833 TI - Effects of bioactive glass particles and their ionic products on intracellular concentrations. AB - Numerous studies have described the bioactive properties of glass particles in the SiO(2)-CaO-Na(2)O-P(2)O(5) system. This kind of material is capable of developing a direct contact with bone through dissolution and physicochemical reactions. We have investigated the influence of bioactive particles, and ionic products from the same particles, on the intracellular concentrations in monocyte cells, which are among the first cells to colonize implantation sites. The only way to access these concentrations and particularly diffusible ionic concentrations (potassium, sodium, and chlorine) is to use cryomethods coupled to electron probe microanalysis. We have paid particular attention to the potassium:sodium ratio, the most sensitive criterion of viability. We have cultured cells with bioactive glass particles and in a conditioned medium obtained from the dissolution of the glass particles in the standard medium. Our study demonstrates that cells cultured in a conditioned medium are more active than cells cultured in a standard medium, or cells exposed to bioactive particles, and particles are more toxic for cells than are ionic products. PMID- 12761834 TI - Use of a biomimetic strategy to engineer bone. AB - Engineering trabecular-like, three-dimensional bone tissue throughout biodegradable polymer scaffolds is a significant challenge. Using a novel processing technique, we have created a biodegradable scaffold with geometry similar to that of trabecular bone. When seeded with bone-marrow cells, new bone tissue, the geometry of which reflected that of the scaffold, was evident throughout the scaffold volume and to a depth of 10 mm. Preseeded scaffolds implanted in non-healing rabbit segmental bone defects allowed new functional bone formation and bony union to be achieved throughout the defects within 8 weeks. This marks the first report of successful three-dimensional bone-tissue engineering repair using autologous marrow cells without the use of supplementary growth factors. We attribute our success to the novel scaffold morphology. PMID- 12761835 TI - Local infusion of FGF-2 enhances bone ingrowth in rabbit chambers in the presence of polyethylene particles. AB - Osseointegration of porous-coated implants during revision arthroplasty procedures is often impeded due to the presence of residual granuloma, particulate debris, and a sclerotic, dysvascular bone bed. We hypothesized that local infusion of recombinant fibroblast growth factor (FGF-2) would increase bone ingrowth in an in vivo model of tissue differentiation in the rabbit tibia in the presence of phagocytosable polyethylene particles. A drug test chamber (DTC) was implanted in the proximal medial tibial metaphysis of mature rabbits unilaterally. The chamber contained a 1x 1 x 5-mm tunnel for tissue ingrowth, and was connected to an osmotic diffusion pump. FGF-2 was infused at dosages of 0, 0.5, 5, 50, or 500 ng/day for a 3-week period, with subsequent harvesting of the ingrown tissue after each 3-week treatment. The effects of ultrahigh molecular weight polyethylene particles (0.5-microm diameter) on tissue ingrowth were determined by adding particles to the chamber at concentrations of 5.8 x 10(11) (low dose) or 1.7 x 10(12) (high dose) particles/mL, with and without infusion of 50 ng/day of FGF for 3 weeks. The tissue forming in the chamber was harvested after each treatment for histologic processing and morphometric analysis of bone ingrowth. Statistical analysis was performed using parametric tests (ANOVA), nonparametric tests (Kruskal-Wallis test) and post hoc tests. In the absence of particles, infusion of 50 ng FGF-2 per day yielded the greatest amount of bone ingrowth. The high dose of particles suppressed bone ingrowth into the chamber, but the low dose particles did not (p = 0.0002, 95% confidence limits = 9.19 18.80). Infusion of 50 ng FGF-2 per day significantly increased net bone formation in the presence of high-dose UHMWPE particles (p = 0.039, 95% confidence limits = 1.41-6.79). There was a trend for decreased numbers of vitronectin-receptor positive (osteoclast-like) cells with the addition of FGF-2, compared to particles alone (p = 0.08). Local delivery of FGF-2 may prove useful in mitigating the adverse effects of wear debris (e.g., in treating early osteolytic lesions), and facilitating osseointegration of revision total joint replacements in situations where the bone bed is suboptimal and residual particles and granulomatous tissue are present. PMID- 12761836 TI - Relative influence of polymer fiber diameter and surface charge on fibrous capsule thickness and vessel density for single-fiber implants. AB - Single polypropylene microfibers plasma-coated with polymers of different surface charge [N,N-dimethylaminoethyl methacrylate (NN) (positive charge), methacrylic acid (MA) (negative charge), and hexafluoropropylene (HF) (neutral)] were implanted in the subcutaneous dorsum of Sprague-Dawley rats for 5-week intervals. Thee groups of fiber diameters were used: (I) 1.0 to 5.9 microm; (II) 6.0 to 10.9 microm; and (III) 11.0 to 15.9 microm. Fibrous capsule thickness and blood-vessel density (number of vessels within 100 microm of the fiber) were assessed in tissue sections in the planes of microfiber cross-sections. Results from a multifactorial analysis of variance demonstrated statistically significant main effects (p < 0.05) for microfiber diameter but not for surface-charge coating. The mean differences in capsule thickness among the microfiber diameter groups were: between groups II and I: 5.4 microm; between groups III and I: 10.2 microm; and between groups III and II: 4.7 microm. The mean differences in capsule thickness among surface-charge coatings were: between MA and NN: 0.7 microm; between MA and HF: 1.4 microm; and between NN and HF: 0.7 microm. Many of the 1.0 to 5.9 microm-in-diameter fibers had no capsule and no sign of a foreign-body reaction. For the vessel density analysis, neither microfiber diameter nor surface-charge coating had a statistically significant effect. Thus the geometric feature of microfiber diameter was more important than was surface charge relative to fibrous capsule formation but not relative to local vessel density. This ranking of the relative influence of design features in relation to tissue response provides useful information for prioritization in biomaterial design. PMID- 12761837 TI - Long-term implantation of zinc-releasing calcium phosphate ceramics in rabbit femora. AB - Zinc is an essential trace element that has stimulatory effects on bone formation. Recently, we developed zinc-releasing calcium phosphate ceramics in order to add the pharmacologic effect of zinc to calcium phosphate ceramics. In our previous study, we showed that the optimum zinc content for promoting bone formation was 0.316 wt %. Therefore a zinc composite ceramic of zinc-containing beta-tricalcium phosphate and hydroxyapatite, with a zinc content of 0.316 wt %, was chosen for long-term implantation. Cylindrical rods of the zinc composite ceramic were implanted in rabbit femora for 2 to 60 weeks. Using computer-aided image analysis, a histomorphometric study was carried out to investigate bone formation and resorption around the implants. The control was a composite ceramic of beta-tricalcium phosphate and hydroxyapatite without zinc. The addition of zinc to the implant demonstrated both favorable and unfavorable effects on bone remodeling. The favorable effect was enhanced bone apposition to the implant surface, demonstrated by a significant increase in intramedullary bone apposition rate at 6 weeks and in cortical bone apposition rate at 24 and 60 weeks (p < 0.05). The unfavorable effect was increased bone resorption, demonstrated by a significant increase in medullary cavity area at 60 weeks (p < 0.05). In order to utilize the favorable effect and avoid the unfavorable effect of zinc, either a reduction in zinc content in the zinc composite ceramic or the selection of implantation sites that do not have excessive exposure to bone marrow are required. PMID- 12761838 TI - Effect of biphasic calcium phosphate on human macrophage functions in vitro. AB - Bioceramics may initiate several and complex biological reactions in host tissues. The cell-biomaterial interaction can determine macrophage activation that may elicit and sustain inflammatory response at the implant site. The current study describes some of the in vitro phenomena regarding the effect of surface reactivity of biphasic calcium phosphate (BCP) granules on human macrophages locomotion and secretion. X-ray diffraction analysis indicated that the synthesized ceramic presented 80% hydroxyapatite and 20% tricalcium phosphate. When BCP was put in contact with human macrophage cells, we observed that cells and BCP granules attached to each other. Cells attached to BCP presented a higher intracellular free Ca(2+) concentration compared with nonattached neighbors and secreted calcium phosphate particles into the medium. Energy dispersive X-ray analysis showed that the secreted particles presented a calcium/phosphorus ratio of 1.64 +/- 0.05, similar to hydroxyapatite. We propose that the secreted particles create a transition zone that allows further macrophage adhesion. PMID- 12761840 TI - Enhancing the vascularization of three-dimensional porous alginate scaffolds by incorporating controlled release basic fibroblast growth factor microspheres. AB - Site-specific delivery of angiogenic growth factors from tissue-engineered devices should provide an efficient means of stimulating localized vessel recruitment to the cell transplants and would ensure cell survival and function. In the present article, we describe the construction of a novel porous alginate scaffold that incorporates tiny poly (lactic-co-glycolic acid) microspheres capable of controlling the release of angiogenic factors, such as basic fibroblast growth factor (bFGF). The microspheres are an integral part of the solid alginate matrix, and their incorporation does not affect the scaffold porosity or pore size. In vitro, bFGF was released from the porous composite scaffolds in a controlled manner and it was biologically active as assessed by its ability to induce the proliferation of cardiac fibroblasts. The controlled delivery of bFGF from the three-dimensional scaffolds accelerated the matrix vascularization after implantation on the mesenteric membrane in rat peritoneum. The number of penetrating capillaries into the bFGF-releasing scaffolds was nearly fourfold higher than into the control scaffolds (those incorporating microspheric BSA and heparin but not bFGF). At day 10 posttransplantation, capillary density in the composite scaffolds was 45 +/- 3/mm(2) and it increased to 70 +/- 7/mm(2) by day 21. The released bFGF induced the formation of large and matured blood vessels, as judged by the massive layer of mural cells surrounding the endothelial cells. The control over bFGF delivery and localizing its effects to areas of need, may aid in the wider application of bFGF in therapeutic angiogenesis as well as in tissue engineering. PMID- 12761841 TI - Novel human endothelial cell-engineered polyurethane biomaterials for cardiovascular biomedical applications. AB - A tri-block coupling-polymer composed of 4,4'-methylenediphenyl diisocyanate and poly (ethylene oxide) (PEO), abbreviated MPEO, was used as the template surface modifying additive (SMA), based on which selected amino acids (lysine, arginine, glycin, and aspartic acid) and RGD peptide were respectively conjugated as functional endgroups of the PEO spacer-arms through sulfonyl chloride-activation routes. After the immobilization of biofunctional factors, the SMA-MPEO derivatives were noncovalently introduced onto the biomedical poly(ether urethane) (PEU) surfaces by physical blending methods. The SMA synthesis and PEU surface modification were monitored and analyzed by nuclear magnetic resonance spectroscopy, attenuated total reflection-infrared spectroscopy, and X-ray photoelectron spectroscopy. The human umbilical vein endothelial cells (HUVECs) were collected and harvested manually by collagenase digestion. The cell culture was performed respectively on the MPEO derivative-modified PEU surfaces and also on the surfaces of the commercially available polystyrene cell-culture plates (TCPS) for control. The cell adhesion rates and cell proliferation rates of the in vitro cultivated HUVEC were measured using flow cytometry. The individual cell viability rates were determined with MTT assay. The cell morphologies of the living HUVECs were investigated by optical inverted microscopy, and more detailed information was acquired from scanning electrical microscopy. The results indicated that the efficacy of SMA functional endgroups was the dominant factor for HUVEC compatibility; the proper-sized PEO spacers (M(w) 2 k) could support and mobilize the functional endgroups, optimizing the surface (interface) environment for the cell growth. As the endgroups of the SMA-MPEO derivatives and the bio-functional factors, the basic amino acids (lysine and arginine) demonstrated similar performances to that of the widely acknowledged cell growth promoter, RGD peptide, which were superior to TCPS. Therefore, these MPEO derivative-modified PEU materials are promising to serve as novel polymeric permanent implants or interventional devices for cardiovascular biomedical applications. PMID- 12761839 TI - Relationship between damage accumulation and mechanical property degradation in cortical bone: microcrack orientation is important. AB - The accumulation of damage and the associated degradation of the mechanical properties of cortical bone are postulated to contribute to age-, disease-, overuse-, and disuse-related skeletal fragilities. Therefore, gaining insight into the relationship between damage and degradation processes is essential in understanding the etiology of skeletal fractures. In investigating this relationship, the damage measure ideally needs to account for the size, the distribution density, and the orientation of microcracks. Existing measures of damage address the size and distribution density of microcracks; however, the orientation of cracks has not been well-investigated. Because the overall orientation of microcracks determines the material axis along which the greatest degradation will be experienced, we hypothesized that the incorporation of the relative orientation between microcracks and loading direction will improve the significance of the relationship between damage accumulation and material property degradation. A three-cycle damage protocol was used to induce tensile damage and to quantify the degradation of the elastic modulus of specimens from human donor femoral cortical bone (a 24-year-old and a 72-year-old man). Microcracks were evaluated by en bloc basic fuchsin staining of specimens after testing. The length (L(i)) and the orientation with respect to the loading direction (beta(i)) of each crack were quantified by a video microscopy system. Three damage measures were quantified for each specimen: the number of linear microcracks (Cr #), the sum of the crack lengths (SigmaL(i)) accounting for the microcrack size alone, and the sum of the projected crack length [SigmaL(Pi) = SigmaL(i)cos(beta(i))] accounting for both crack size and orientation. Inclusion of the orientation parameter improved the coefficient of determination between damage accumulation and the degradation of the elastic modulus: the coefficient of determination of the sum of the projected crack length (R(2) = 0.239) was 60% greater than that of the sum the projected crack length (R(2) = 0.149) and 33% greater than that of the number of linear microcracks (R(2) = 0.180). We conclude that microcrack orientation is an essential physical variable in the relationship between damage accumulation and degradation of mechanical properties of cortical bone tissue. PMID- 12761842 TI - Engineering integrin-specific surfaces with a triple-helical collagen-mimetic peptide. AB - Integrin-mediated cell adhesion to extracellular matrix proteins anchors cells and triggers signals that direct cell function. The integrin alpha(2)beta(1) recognizes the glycine-phenylalanine-hydroxyproline-glycine-glutamate-arginine (GFOGER) motif in residues 502-507 of the alpha(1)(I) chain of type I collagen. Integrin recognition is entirely dependent on the triple-helical conformation of the ligand similar to that of native collagen. This study focuses on engineering alpha(2)beta(1)-specific bioadhesive surfaces by immobilizing a triple-helical collagen-mimetic peptide incorporating the GFOGER binding sequence onto model nonadhesive substrates. Circular dichroism spectroscopy verified that this peptide adopts a stable triple-helical conformation in solution. Passively adsorbed GFOGER-peptide exhibited dose-dependent HT1080 cell adhesion and spreading comparable to that observed on type I collagen. Subsequent antibody blocking conditions verified the involvement of integrin alpha(2)beta(1) in these adhesion events. Focal adhesion formation was observed by immunofluorescent staining for alpha(2)beta(1) and vinculin on MC3T3-E1 cells. Model functionalized surfaces then were engineered using three complementary peptide-tethering schemes. These peptide-functionalized substrates supported alpha(2)beta(1) mediated cell adhesion and focal adhesion assembly. Our results suggest that this peptide is active in an immobilized conformation and may be applied as a surface modification agent to promote alpha(2)beta(1)-specific cell adhesion. Engineering surfaces that specifically target certain integrin-ligand interactions and signaling cascades provides a biomolecular strategy for optimizing cellular responses in biomaterials and tissue engineering applications. PMID- 12761843 TI - Biodegradation of polyurethane under fatigue loading. AB - A method utilizing expansion of a diaphragm-type film specimen was developed to study in vitro biodegradation of poly(etherurethane urea) (PEUU) under conditions of dynamic loading (fatigue). A finite element model was used to describe the strain state, which ranged from uniaxial at the edges of the film to balanced biaxial tensile strain at the center. During testing, the film was exposed to a H(2)O(2)/CoCl(2) solution, which simulated in vivo oxidative biodegradation of PEUU. The extent of chemical degradation was determined by infrared analysis. Physical damage of the film surface was characterized by optical microscopy and scanning electron microscopy. Dynamic loading did not affect the rate of degradation relative to unstressed and constant stress (creep) controls in regions of the film that experienced primarily uniaxial fatigue; however, degradation was accelerated in regions that experienced balanced biaxial or almost balanced biaxial fatigue. It was concluded that the combination of dynamic loading and biaxial tensile strain accelerated oxidative degradation in this system. Chemical degradation produced a brittle surface layer that was marked by numerous pits and dimples. Physical damage of the surface in the form of cracking occurred only in fatigue experiments. Cracking was not observed in unstressed or creep tests. Cracks initiated at the dimples produced by chemical degradation, and propagated in a direction that was determined by the strain state. PMID- 12761844 TI - Platelet-derived growth factor signaling as a cue of the epithelial-mesenchymal interaction required for anuran skin metamorphosis. AB - The anuran remodels the larval skin into the adult counterpart during metamorphosis. The construction of the precursor of adult epidermis (preadult epidermis) in Xenopus laevis larvae was coordinated with the development of the secondary connective tissue (s-ct) underneath the basement membrane, suggesting that the epithelial-mesenchymal interaction plays a critical role in the metamorphic conversion of the larval skin. mRNAs of platelet-derived growth factor A (PDGF-A) and PDGF receptor (PDGFR) -alpha were markedly up-regulated in the skin during spontaneous and thyroid hormone (TH) -induced metamorphosis. In situ hybridization experiments identified preadult epidermal basal cells and fibroblasts in developing subepidermal connective tissues at the late prometamorphic stage as PDGF-A and PDGFR-alpha mRNA-expressing cells, respectively. We developed an in vitro model of larval skin that was remodeled to the adult skin under the influence of TH. The presence of either of AG1296, a specific inhibitor of PDGFR tyrosine kinase autophosphorylation, or an excess of recombinant proteins of the soluble extracellular domain of PDGFR-alpha inhibited the following TH-induced processes, the proliferation of adult basal cells, the terminal differentiation of adult basal cells, and the activation of subepidermal fibroblasts. However, the inhibitors did not inhibit the TH-induced proliferation of preadult basal cells. We concluded that PDGF/PDGFR signaling is one of the prime cues in the epithelial-mesenchymal interaction required for the metamorphic skin remodeling. PMID- 12761845 TI - Localisation of specific heparan sulfate proteoglycans during the proliferative phase of brain development. AB - Early brain development is characterised by the proliferation of neural precursor cells. Several families of signalling molecules such as the fibroblast growth factors (FGFs) and Wnts are known to play important roles in this early phase of brain development. Accumulating evidence demonstrates that signalling of these molecules requires the presence of heparan sulfate chains attached to a proteoglycan core protein (HSPG). However, the specific identity of the HSPG components in the developing brain is unknown. To determine which HSPGs might be involved at this early phase, we analysed the expression of the major cell surface HSPG families in the developing brain at a time of most active proliferation. Syndecan-1 and glypican-4 were the most highly expressed in the developing brain during the time of peak proliferation and localise to ventricular regions of the brain, where the precursor cells are proliferating. Syndecan-4, although less abundant, also localises to cells in the ventricular zone. We have also examined HSPG involvement in brain development using cultures of embryonic neural precursor cells. We find that FGF2 stimulation of proliferation is inhibited in the presence of sodium chlorate, an inhibitor of heparan sulfate synthesis, and is rescued by addition of exogenous heparan sulfate. These data support a requirement for heparan sulfate in FGF signalling for proliferation of brain precursor cells. The expression of these specific HSPGs within the proliferative zone of the brain suggests that they may be involved in regulation of early brain development, such as FGF-stimulated proliferation. PMID- 12761846 TI - Differential expression of angiogenic and vasodilatory factors by invasive trophoblast giant cells depending on depth of invasion. AB - The uterine bed undergoes remarkable changes during pregnancy, including proliferation and decidualization of the uterine stroma and remodeling and angiogenesis of the maternal vasculature. Fetal-derived trophoblast giant cells invade into the uterus where they gain access to the maternal blood circulation to ensure sufficient nutrient supply of the embryo. In serial sections through early- to mid-gestation conceptuses, we have determined the exact distance of trophoblast invasion and the expression of angiogenic, vasodilatory, and anticoagulative factors that are likely to influence remodeling and redirection of the maternal circulatory system. Trophoblast derivatives were detected at a distance as far as approximately 300 microm from the placental border, where they are allocated exclusively along the mid-line of the decidua. The farthest invading cells characteristically expressed proliferin and proliferin-related protein, hormones that affect endothelial cell migration and vascularization. Occasionally, these cells replaced the normal vascular endothelium and acquired a "pseudo-endothelial" shape. Complete vascular disintegration was observed 50-80 microm outside of the placental border where maternal blood was entirely lined by a trophoblast giant cell-derived network of blood sinuses. This transition in blood space lining correlated with trophoblast expression of various vasodilatory and anticoagulative factors that are likely to promote blood flow toward the placenta. Analysis of teratocarcinoma-like tumors demonstrated that trophoblast giant cell-induced promotion and redirection of blood flow is not restricted to the uterine environment. These results show that trophoblast giant cells have the intrinsic capacity to attract and increase blood flow and to gradually displace the vascular endothelium resulting in the formation of canals entirely lined by trophoblast cells. PMID- 12761847 TI - HOXB13 homeodomain protein is cytoplasmic throughout fetal skin development. AB - Substantial evidence suggests that HOX homeobox genes regulate aspects of body development, including hair formation. We initially isolated the HOXB13 gene from human fetal skin in experiments designed to identify candidate genes that regulate scarless fetal wound healing. Although the HOX homeodomain proteins have been proposed to function as transcription factors, we have demonstrated previously that substantial fractions of the HOXB6 and HOXB4 proteins are localized to the cytoplasm throughout epidermal development. The purpose of the current study was to identify HOXB13 protein expression patterns in developing skin to elucidate potential mechanisms by which this protein might regulate aspects of tissue development and healing. HOXB13 protein expression was detected throughout the developing epidermis, with weaker signal observed in the early developing dermis. Epidermal HOXB13 signal was detected over the entire body surface, but surprisingly, essentially all of the signal was cytoplasmic in developing skin. Low-level HOXB13 protein expression was detected in adult skin and within the telogen hair follicle, and a portion of the residual signal in adult epidermis was nuclear. Expression in hyperproliferative skin conditions remained cytoplasmic with the exception of epidermis associated with Kaposi's sarcoma, which showed strong HOXB13 expression that was partially localized to the nucleus. PMID- 12761848 TI - Expression and function of FGF10 in mammalian inner ear development. AB - We have investigated the expression of FGF10 during ear development and the effect of an FGF10 null mutation on ear development. Our in situ hybridization data reveal expression of FGF10 in all three canal crista sensory epithelia and the cochlea anlage as well as all sensory neurons at embryonic day 11.5 (E11.5). Older embryos (E18.5) displayed strong graded expression in all sensory epithelia. FGF10 null mutants show complete agenesis of the posterior canal crista and the posterior canal. The posterior canal sensory neurons form initially and project rather normally by E11.5, but they disappear within 2 days. FGF10 null mutants have no posterior canal system at E18.5. In addition, these mutants have deformations of the anterior and horizontal cristae, reduced formation of the anterior and horizontal canals, as well as altered position of the remaining sensory epithelia with respect to the utricle. Hair cells form but some have defects in their cilia formation. No defects were detected in the organ of Corti at the cellular level. Together these data suggest that FGF10 plays a major role in ear morphogenesis. Most of these data are consistent with earlier findings on a null mutation in FGFR2b, one of FGF10's main receptors. PMID- 12761849 TI - EGF-stimulated signaling by means of PI3K, PLCgamma1, and PKC isozymes regulates branching morphogenesis of the fetal mouse submandibular gland. AB - Branching morphogenesis of fetal mouse submandibular glands (SMGs) partly depends on the epidermal growth factor (EGF) receptor that triggers at least three intracellular signaling pathways involving (1) the mitogen-activated protein kinases ERK-1/2, (2) phospholipase Cgamma1 (PLCgamma1), and (3) phosphatidylinositol-3-kinase (PI3K). PLCgamma1 directly activates protein kinase C (PKC) isozymes; PI3K stimulates protein kinase B (PKB, also known as Akt), which ultimately activates PKCs and other proteins. We reported that the pattern of phosphorylation of ERK-1/2 in response to EGF in SMGs varies with fetal age and that blockade of EGF-stimulated ERK-1/2 signaling partially inhibits branching (Kashimata et al. [2000] Dev. Biol. 220:183-196). Here, we report on components of the PLCgamma1, PI3K, and PKC families of signaling molecules in fetal SMGs from the 13th day of gestation to postnatal ages. Western blotting revealed that (1) PLCgamma1 is present from E13 to E18 but drops off precipitously to negligible levels on the day of birth and thereafter, and (2) PI3K, PKB(Akt), and several PKC isozymes are expressed from E13 onward through adult life. Both PLCgamma1 and PI3K are phosphorylated in response to EGF. Inhibition of PI3K by LY294002 inhibited EGF-stimulated branching, but inhibition of PLCgamma1 by U73122 had no effect. Western blotting showed that the concentrations of 8 PKC isozymes vary with age in the fetal and postnatal SMG. However, general inhibition of PKCs by Calphostin C or specific inhibition of PKCalpha or of PKCepsilon by Go6976 or Ro-32-0432, respectively, increased EGF stimulated branching. Calphostin C also increased EGF-stimulated phosphorylation of ERK-1/2. These findings indicate that signaling from the EGF receptor in the fetal mouse SMG varies with development and triggers stimulatory effects by means of ERK-1/2 and PI3K but inhibitory effects by means of PKC isozymes. PMID- 12761851 TI - Analysis of regulatory elements of E-cadherin with reporter gene constructs in transgenic mouse embryos. AB - Proper regulation of E-cadherin-mediated cell adhesion is important during early embryonic development and in organogenesis. In mice, E-cadherin is expressed from the fertilized egg onward and becomes down-regulated during gastrulation in mesoderm and its derivatives, but its expression is maintained in all epithelia. E-cadherin promoter analyses led to the identification of binding sites for two transcriptional repressors, Snail and SIP1, which are able to mediate down regulation in vitro, but little is known about the regulatory elements that govern E-cadherin transcriptional activity in vivo. Here, we compared the developmentally regulated expression of a series of lacZ-reporter transgenes fused to different sequences of the murine E-cadherin gene between -6 kb, including the promoter, and +16 kb, covering one third of intron 2. Four different segments with distinct regulatory properties were identified. The promoter fragment from +0.1 to -1.5 kb remains inactive in most cases but occasionally induces ectopic expression in mesodermal tissues, although it contains binding sites for the repressors Snail and SIP1. This promoter fragment also lacks positive elements needed for the activation of transcription in ectoderm and endoderm. Sequences from -1.5 to -6 kb harbor regulatory elements for brain-specific expression and, in addition, insulator or silencer elements, because they are consistently inactive in the mesoderm. Only if sequences from +0.1 to +11 kb are combined with the promoter fragments is E-cadherin-specific transgene expression observed in endoderm and certain epithelia. Sequences between +11 and +16 kb contain cis-active elements that generally enhance transcription. Our analyses show that E-cadherin expression is governed by a complex interplay of multiple regulatory regions dispersed throughout large parts of the locus. PMID- 12761850 TI - Thyroid transcription factor (TTF) -1 regulates the expression of midkine (MK) during lung morphogenesis. AB - Midkine (MK) is a 13-kDa heparin-binding growth factor that is thought to mediate developmental processes, including vasculogenesis, cell migration, and proliferation in various organs. To determine whether MK plays a role during lung morphogenesis, immunostaining for MK was assessed in mouse lung from embryonic day (E) 13 to postnatal day (PN) 24. MK was detected in mesenchymal and respiratory epithelial cells of the peripheral mouse lung from E13.0 to E15.5. From E18.5 to PN1, MK was observed primarily in epithelial cells lining conducting airways and peripheral lung saccules. By PN10, expression was no longer observed in respiratory epithelial cells but was readily detected in small blood vessels in the alveolar region of the lung. Although most respiratory epithelial cells uniformly expressed MK before E13.0, MK was restricted to subsets of cells by E18.5, colocalizing with the Clara cell secretory protein (CCSP) marker in conducting airways and with pro-SPC, a marker specific for alveolar type II pneumocytes. By PN10, MK was not detected in respiratory epithelial cells of the conducting airways and was closely associated with capillary networks. The sites of intense MK staining in the respiratory epithelial cells correlated with sites of expression of thyroid transcription factor (TTF) -1, a transcription factor regulating formation and gene expression in the lung parenchyma. TTF-1 enhanced transcription of the mouse MK gene promoter, acting on TTF-1 regulatory elements located in the 5'-region of the gene. Furthermore, MK expression was not detected in lungs of TTF-1 null mice. TTF-1 regulates expression of MK in the lung. The temporal/spatial distribution of midkine is consistent with a potential role in paracrine signaling during lung morphogenesis. PMID- 12761852 TI - Dual mechanisms governing muscle cell death in tadpole tail during amphibian metamorphosis. AB - The tadpole tail, which is twice as long as the body, is induced to resorb completely by thyroid hormone within several days during the anuran metamorphosis. To investigate the underlying mechanism, we undertook two approaches. First, we examined the effect of dominant-negative thyroid hormone receptor (DNTR) on muscle cell death in vitro. The overexpression of DNTR suppressed the death of a tail-derived myoblastic cell line induced by thyroid hormone. Second, tadpole tails were injected with a reporter gene and the DNTR expression construct, and the reporter gene expression in muscle cells was followed during the spontaneous metamorphosis. DNTR overexpression inhibited a decrease of the reporter gene expression that began at stage 57 in the control tadpoles but only delayed massive muscle cell death at stage 63 when tails shrink very rapidly. Some remained even a few weeks after the metamorphosis, although most DNTR-overexpressing cells died by the end of the metamorphosis. These results led us to propose that thyroid hormone induces the suicide of muscle cells (the cell-autonomous death) in the tail between stage 57 and 62 and that both the murder and suicide mechanisms execute muscle cell death in stage 62-64 to remove muscle promptly and completely. PMID- 12761853 TI - Expression and neural control of follistatin versus myostatin genes during regeneration of mouse soleus. AB - Follistatin and myostatin are two secreted proteins involved in the control of muscle mass during development. These two proteins have opposite effects on muscle growth, as documented by genetic models. The aims of this work were to analyze in mouse, by using in situ hybridization, the spatial and temporal expression patterns of follistatin and myostatin mRNAs during soleus regeneration after cardiotoxin injury, and to investigate the influence of innervation on the accumulation of these two transcripts. Follistatin transcripts could be detected in activated satellite cells as early as the first stages of regeneration and were transiently expressed in forming myotubes. In contrast, myostatin mRNAs accumulated persistently throughout the regeneration process as well as in adult control soleus. Denervation significantly affected both follistatin and myostatin transcript accumulation, but in opposite ways. Muscle denervation persistently reduced the levels of myostatin transcripts as early as the young myotube stage, whereas the levels of follistatin mRNA were strongly increased in the small myotubes in the late stages of regeneration. These results are discussed with regard to the potential functions of both follistatin, as a positive regulator of muscle differentiation, and myostatin, as a negative regulator of skeletal muscle growth. We suggest that the belated up-regulation of the follistatin mRNA level in the small myotubes of the regenerating soleus as well as the down-regulation of the myostatin transcript level after denervation contribute to the differentiation process in denervated regenerating muscle. PMID- 12761854 TI - Molecular structure and developmental expression of three muscle-type troponin T genes in zebrafish. AB - Troponin T (Tnnt), a troponin component, interacts with tropomyosin and is crucial to the regulation of striated muscle contraction. To gain insight into the molecular evolution and developmental regulation of Tnnt gene (Tnnt) in lower vertebrates, zebrafish Tnnt1 (slow Tnnt), Tnnt2 (cardiac Tnnt), and Tnnt3b (fast Tnnt isoform b) were characterized. The polypeptides of zebrafish Tnnt1, Tnnt2, and Tnnt3b were conserved in the central tropomyosin- and C-terminal troponin I binding domains. However, the N-terminal hypervariable regions were highly extended and rich in glutamic acid in polypeptides of Tnnt1 and Tnnt2, but not Tnnt3b. The Tnnt2 and Tnnt3b contain introns, whereas Tnnt1 is intron-free. During development, large to small, alternatively spliced variants were detected in Tnnt2, but not in Tnnt1 or Tnnt3. Whole-mount in situ hybridization showed zebrafish Tnnt1 and Tnnt2 are activated during early somitogenesis (10 hr postfertilization, hpf) and cardiogenesis (14 hpf), respectively, but Tnnt3b is not activated until middle somitogenesis (18 hpf). Tnnt2 and Tnnt3b expression was cardiac- and fast-muscle specific, but Tnnt1 was expressed in both slow and fast muscles. We propose that three, distinct, muscle-type Tnnt evolved after the divergence of fish and deuterostome invertebrates. In zebrafish, the developmental regulation of Tnnt during somitogenesis and cardiogenesis is more restricted and simpler than in tetrapods. These new findings may provide insight into the developmental regulation and molecular evolution of vertebrate Tnnt. PMID- 12761855 TI - Intracellular calcium plays an essential role in cardiac development. AB - Intracellular calcium signaling plays an essential role in cardiac physiology and modulates cardiac gene expression. However, the role that intracellular calcium signaling plays during cardiac development is not known. To address this issue, we examined the effects of altered intracellular calcium levels on cardiac morphogenesis. In acutely cultured mouse embryos, L-type calcium channel blockade decreased resting intracellular calcium levels and inhibited calcium transients. Embryos cultured at embryonic day (E) 7.5-8.5 in the presence of the L-type calcium channel blockers nifedipine and verapamil developed hearts that had a large left ventricle, lacked a right ventricle and had a long, thin outflow tract. If embryos were cultured at E7.5, calcium channel blockade also induced an abnormal, anterior cardiac loop. These alterations in development were not due to altered cardiac function, as heart rates at the end of the culture period were not affected by calcium channel blockade and blood flow was observed. Treatment with nifedipine altered the mRNA expression of the transcription factor Gata4, which was absent in the developing ventricles, and the sarcomeric protein Mylpc (myosin light chain 2V), which was decreased distal to the left ventricle and was absent at the site of the developing right ventricle. In contrast, the expression pattern of other cardiac transcription factor (Hand1, Hand2, Mef2c, Nkx2-5) and cytoskeletal protein (Myhca, Tagln) mRNA did not change with calcium channel blockade. These data demonstrate that proper intracellular calcium signaling is essential for normal cardiac looping, gene expression, and organ development. PMID- 12761856 TI - Phenotype of the zebrafish masterblind (mbl) mutant is dependent on genetic background. AB - The zebrafish masterblind (mbl) mutant is characterized by the lack of olfactory placodes and optic vesicles, reduced telencephalon, an expanded epiphysis (Heisenberg et al. [1996] Development 123:191-203), and enlarged jaw. To understand the cellular events giving rise to the olfactory placode defect of this mutant, we examined the expression pattern of the distal-less-3 (dlx3) gene in mbl. In the mutant, dlx3, which is normally expressed in the developing nose and ear, showed reduced expression in the olfactory placode field, but normal expression in the developing ear. To determine whether the loss of dlx3 expression was due to cell loss, we assayed cell death by using TUNEL labeling. Although cell death in the mutant was not concentrated in the region of dlx3 expression, there was increased cell death in the forebrain, epiphysis, and jaw region, as compared with that in wild-type controls. This cell death phenotype was cyclical in nature, showing an increase and decrease in cell death on a roughly 24-hr cycle. Further analysis showed that this cyclical phenotype was specific to the genetic background. The severity of the mbl phenotype, including cell death, expanded epiphysis, and enlarged jaw, decreased when the mutation was moved from the original "TL" background to the "AB" background. Thus, the severity of developmental defects in the mbl mutant is strongly dependent on genetic background. We examined the contribution of cell death to the morphologic defects of mbl by blocking cell death by using zVADfmk, a known caspase inhibitor. We found that this treatment partially rescued the expanded jaw defect and that this rescue was dependent on the genetic background. Therefore, the mbl mutant phenotypes result, in part, from genetic background effects that alter the pattern of programmed cell death early in development. PMID- 12761857 TI - Folic acid and homocysteine affect neural crest and neuroepithelial cell outgrowth and differentiation in vitro. AB - The beneficial effect of additional folic acid in the periconceptional period to prevent neural tube defects, orofacial clefts, and conotruncal heart defects in the offspring has been shown. Folate shortage results in homocysteine accumulation. Elevated levels of homocysteine have been related to neural tube defects. We studied the behavior of neuroepithelial cells and cranial and cardiac neural crest cells in vitro. Neural tube explants were cultured for 24 and 48 hr in medium after addition of folic acid and/or homocysteine. Folic acid addition increased neuroepithelial cell outgrowth and increased neural crest cell differentiation into nerve and smooth muscle cells. Addition of homocysteine increased neural crest cell outgrowth and migration from the neural tube and inhibited neural crest cell differentiation. Our findings suggest that neural tube defects caused by folate deficiency and hyperhomocysteinemia develop due to increased neuroepithelial to neural crest cell transformation. This increased transformation leads to a shortage of neuroepithelial cells in the neural tube. Defects in orofacial and conotruncal development are explained by abnormal differentiation of neural crest cells in the presence of high homocysteine concentrations. Our findings supports a critical role for folic acid and homocysteine in the development of neural tube defects and neural crest related heart malformations. PMID- 12761858 TI - Differential expression of beta3 integrin gene in chick and mouse cranial neural crest cells. AB - RNA in situ hybridization on early chicken embryos revealed that the beta3 integrin gene started to be expressed after Hamburger and Hamilton (HH) stage 6 in the presumptive epidermis adjacent to the neural plate, before closure of the neural tube. The beta3 integrin gene was also strongly expressed in cephalic neural crest cells at the same stage in which they begin their migration but disappeared progressively in these cells along the route they take to the branchial arches. The gene was weakly expressed in the differentiating cranial neural crest cells. The alphaVbeta3 integrin protein complex was also mainly detected in the migratory cephalic neural crest cells. However, during early mouse embryogenesis and in contrast to the chick, the beta3 integrin gene was expressed in the foregut diverticulum and in the heart and not in the cephalic neural crest cells. Therefore, the difference in the beta3 integrin expression suggests that mouse and chicken cranial neural crest cells may have distinct integrin requirements during their ontogenesis. PMID- 12761859 TI - To be glial or not-how glial are the precursors of neurons in development and adulthood? PMID- 12761860 TI - What is a glial cell? PMID- 12761861 TI - Precursors of neurons, neuroglia, and ependymal cells in the CNS: what are they? Where are they from? How do they get where they are going? AB - Neurons, neuroglia (astrocytes and oligodendrocytes), and ependymal cells are three distinct categories of neural cells in the central nervous system. In the mature brain and spinal cord, the classical histological criteria define these cells by their microscopic structure very well. During development, the precursors for all of these cells reside within the epithelium of the neural plate and its successor, the neural tube. These precursor cells are the undifferentiated, primitive neuroepithelium of the classical literature. As the cerebral vesicles enlarge and their walls thicken, the primitive neuroepithelial cells elongate, maintaining a radial orientation until they migrate. Although many, but not all, of these cells span the extent of the ventricular wall, they are the precursors of neurons, neuroglia, and ependymal cells. Thus, it is useful to retain their classical designation as primitive neuroepithelial cells and to treat them as neural precursor cells. Neural precursor cells are neither neuroglia nor neurons. It is not appropriate to call them radial glial cells anymore than it is to call them radial neuronal cells. The term "radial glia" has long been used to describe the mature, elongated astrocytes, represented by Bergmann cells in the cerebellum and Muller cells in the retina. Inevitably, during development, transitional forms between neural precursor cells and the neurons, neuroglia, and ependymal cells will occur. Such transitional cells are known as neuroblasts, glioblasts, or ependymoblasts, even though they may be postmitotic. Alternative terms are "immature neurons," "immature neuroglia," and "immature ependymal cells." The migration of many neural precursor cells is accomplished by translocation rather than free cellular locomotion. There is both direct and indirect evidence to document the translocation of the nuclear/perikaryal/somal complex through the leading process of primitive neuroepithelial cells. This is conspicuous in the neocortex, where the discrete radial arrangement of pyramidal cells may result from translocation of neuroblasts, while their leading processes still contact the pial surface. Migration by translocation occurs throughout the CNS. GLIA 43:6-18, 2003. PMID- 12761862 TI - Elusive radial glial cells: historical and evolutionary perspective. AB - Since the discovery of radial glial cells in the human fetal forebrain, this specialized cellular population has been identified in most regions of the vertebrate brain during restricted developmental periods. However, their size, longevity, and significance for guiding migrating neurons have increased with the evolutionary expansion of the mammalian neocortex, reaching a peak in the gyrencephalic human forebrain. The phenotypic distinction of radial glial cells from the more specialized neuronal progenitors in the proliferative zones and from the migrating neurons in the intermediate zone of the primate fetal forebrain, based initially on morphological criteria, has been supported by their ultrastructural, molecular, and physiological characteristics. In addition, modern in vivo and in vitro approaches revealed that these specialized embryonic cells can also generate neuronal cell lines, which either immediately, or after several divisions, migrate along radial shaft processes of the mother cells that span the expanding and convoluted cerebral wall. The multiple functions of radial glial cells and their species-specific adaptations indicate a pivotal role in evolution, development, and pathology of the cerebral neocortex. PMID- 12761863 TI - Radial glia and somal translocation of radial neurons in the developing cerebral cortex. AB - A series of recent studies have demonstrated that radial glia are neural precursors in the developing cerebral cortex. These studies have further implied that these cells are the sole precursor constituents of the dorsal forebrain ventricular zone that generate the projection neurons of the cortex. In view of these new findings, this review discusses radial neurons, a progeny of cortical neurons that are generated by radial glia and adopt somal translocation as the mode of migration. PMID- 12761864 TI - Radial glia diversity: a matter of cell fate. AB - Early in development of the central nervous system, radial glial cells arise from the neuroepithelial cells lining the ventricles around the time that neurons begin to appear. The transition of neuroepithelial cells to radial glia is accompanied by a series of structural and functional changes, including the appearance of "glial" features, as well as the appearance of new signaling molecules and junctional proteins. However, not all radial glia are alike. Radial glial lineages appear to be heterogeneous both within and across different brain regions. Subtypes of neurogenic radial glia within the cortex, for example, may have restricted potential in terms of the cell types they are able to generate. Radial glia located in different brain regions also differ in their expression of growth factors, a diverse number of transcription factors, and the cell types they generate, suggesting that they are involved in regionalization of the developing nervous system in several aspects. These findings highlight the important but complex role of radial glia as participants in key steps of brain development. PMID- 12761865 TI - Signaling to and from radial glia. AB - Radial glia represent the major glial cell type in the developing CNS and perform many essential functions, which range from acting as neural precursors to providing physical substrates for newborn neurons to migrate on. Previous work has shown that cell-cell signaling is important for the development of the radial glial phenotype. In particular, signals from newborn neurons appear to contribute significantly to the formation of this cell type. In addition, radial glia may be involved in reciprocal signaling roles that contribute to regional patterning and differentiation in the developing CNS. PMID- 12761866 TI - Region-specific effects of glia on neuronal induction and differentiation with a focus on dopaminergic neurons. AB - Radial glia (RG) are the first glial cell type to appear in the nervous system. Their broad distribution and apparent similarity hide important brain region specific differences that are likely to be essential for development. However, recent evidence supports the stimulating concept that in addition to their classical function as neuroblast guides, RG are neuronal precursors (Malatesta et al. Development 127:5253-5263, 2000; Miyata et al. Neuron 31:727-741, 2001; Noctor et al. Nature 409:714-720, 2001; Skogh et al. Mol Cell Neurosci 17:811 820, 2001). We propose that RG not only generate and guide newborn neurons, but could also instruct their own neuronal progeny to adopt appropriate region specific phenotypes. PMID- 12761868 TI - Astrocytes as stem cells: nomenclature, phenotype, and translation. AB - Recently discovered multipotent astrocytic stem cells are discussed in light of current nomenclature for glial precursor and lineage-associated cells in the developing, postnatal, and adult mammalian brain. Defining the phenotype of any immature cell in the nervous system is a challenge, and a position is stated that includes the need for categorizing cells within a continuum of differentiation potential. The possibility for dedifferentiating glial cells into clonogenic stem like cells offers numerous possibilities for translating knowledge and technology from this subfield of stem cell biology to regenerative medicine. Along with the need for developing a new lexicon for defining the cellular players that contribute to the generation of glia and neurons in the developing and mature central nervous system, the relationships also need to be established among potency, repopulation attempts, and tumorigenesis of cells meeting the criteria of glial stem cells. Finally, it is possible that understanding the normal differentiation, de- and transdifferentiation potential of glial stem-like cells in the mature central nervous system will provide insights into the possible use of these cells, or biogenic factors associated with their growth and differentiation, in therapeutic approaches for a variety of neurological disorders. PMID- 12761867 TI - Gliogenic and neurogenic progenitors of the subventricular zone: who are they, where did they come from, and where are they going? AB - The subventricular zone (SVZ) of the perinatal forebrain gives rise to both neurons and glia. The mechanisms governing the phenotypic specification of progenitors within this heterogeneous germinal zone are unclear. However, the characterization of subpopulations of SVZ cells has given us a better understanding of the basic architecture of the SVZ and presents us with the opportunity to ask more detailed questions regarding phenotype specification and cell fate. Recent work demonstrating the embryonic origins of SVZ cells is summarized, and a model describing the formation of the perinatal SVZ, noting contributions of cells from pallial as well as subpallial germinal zones, is presented. We further address differences among classes of SVZ cells based on molecular profile, phenotype, and migration behavior and present a model summarizing the organization of perinatal SVZ cells along coronal, sagittal, and horizontal axes. A detailed description of the SVZ in the adult, outlining classes of cells based on morphology, molecular profile, and proliferative behavior, was recently reported by Doetsch et al. (Proc Natl Acad Sci USA 93:14895-14900, 1997). Potential relationships among cells within the perinatal and adult SVZ will be discussed. GLIA 43:52-61, 2003. PMID- 12761869 TI - Potential of Muller glia to become neurogenic retinal progenitor cells. AB - The possibility of neural regeneration has gained credence with the identification of neural stem cells seeded within different regions of the adult central nervous system (CNS). Recently, this possibility has received an additional boost from reports that glia, the support cells of the CNS, might provide a source of neural regeneration. We review some of our findings that Muller glia in the chicken retina are a source of proliferating progenitors that can generate neurons. These Muller cells are fully differentiated glial cells that serve functions ascribed to this cell type. In response to damage or exogenous growth factors, Muller glia dedifferentiate, proliferate, express combinations of transcription factors normally expressed by embryonic retinal progenitors, and produce new neurons and glia. In light of these data, the potential of Muller glia as a source of neural regeneration in the retina of nonavian species, namely humans, seems an avenue of investigation that warrants serious consideration. PMID- 12761870 TI - Potential role of radial glia in adult neurogenesis of teleost fish. AB - Persistence of radial glia within the adult central nervous system is a widespread phenomenon among fish. Based on a series of studies in the teleost species Apteronotus leptorhynchus, we propose that one function of this persistence is the involvement of radial glia in adult neurogenesis, i.e., the generation and further development of new neurons in the adult central nervous system. In particular, evidence has been obtained for the involvement of radial glia in the guidance of migrating young neurons in both the intact and the regenerating brain; for a possible role as precursor cells from which new neurons arise; and for its role as a source of trophic substances promoting the generation, differentiation, and/or survival of new neurons. These functions contribute not only to the potential of the intact brain to generate new neurons continuously, and of the injured brain to replace damaged cells by newly generated ones, but they also provide an essential part of the cellular substrate of behavioral plasticity. PMID- 12761871 TI - GFAP: functional implications gleaned from studies of genetically engineered mice. AB - GFAP is the major intermediate filament of mature astrocytes, and its relatively specific expression in these cells suggests an important function. To study the role of the GFAP gene, mice have been created carrying null alleles (no protein), modified alleles (altered protein), or added wild type alleles (elevated protein). Surprisingly, absence of GFAP has relatively subtle effects on development. On the other hand, over-expression can be lethal, and led to the discovery that GFAP coding mutations are responsible for most cases of Alexander disease, a devastating neurodegenerative disorder. Here we review what the various GFAP mouse models reveal about GFAP and astrocyte function. PMID- 12761872 TI - Astrocytes as support for axonal regeneration in the central nervous system of mammals. AB - Reactive astrocytes are one of the main impediments for axonal regeneration in the central nervous system of mammals. Using mice KO for GFAP and vimentin, we show that reinnervation occurs after an hemisection of the spinal cord, mainly through sprouting of controlateral intact serotoninergic and cortico-spinal axons, thanks to the absence of glial reactivity. This reinnervation is paralleled by the restoration of impaired locomotion of the ipselateral hindleg. Future applications to spinal cord injured patients are discussed. PMID- 12761873 TI - Molecular cloning, expression, and characterization of a cDNA encoding the arylphorin-like hexameric storage protein from the mulberry longicorn beetle, Apriona germari. AB - We describe here the cloning, expression, and characterization of a cDNA encoding the arylphorin-like hexameric storage protein from the mulberry longicorn beetle, Apriona germari (Coleoptera, Cerambycidae). The complete cDNA sequence of A. germari hexamerin (AgeHex) is comprised of 2,160 bp with 720 amino acid residues. The deduced protein sequence of AgeHex is most similar to Tenebrio molitor hexamerin2 (65.3%). Phylogenetic analysis further confirmed the AgeHex is more closely related to T. molitor hexmerin2 and T. molitor early-staged encapsulation inducing protein than to the other insect storage proteins. Southern blot analysis suggested the presence of A. germari hexamerin gene as a single copy and Northern blot analysis confirmed fat body-specific expression at the transcriptional level. The cDNA encoding AgeHex was expressed as a 80-kDa protein in the baculovirus-infected insect cells. Western blot analysis using the polyclonal antiserum against recombinant AgeHex indicated that the AgeHex corresponds to storage protein 2 (SP2) present in the A. germari larval hemolymph. PMID- 12761874 TI - Triacylglycerol and phospholipid fatty acids of the silverleaf whitefly: composition and biosynthesis. AB - The identification and composition of the fatty acids of the major lipid classes (triacylglycerols and phospholipids) within Bemisia argentifolii Bellows and Perring (Homoptera: Aleyrodidae) nymphs were determined. Comparisons were made to fatty acids from the internal lipids of B. argentifolii adults. The fatty acids, as ester derivatives, were analyzed by capillary gas chromatography (CGC) and CGC mass spectrometry (MS). All lipid classes contained variable distributions of eight fatty acids: the saturated fatty acids, myristic acid (14:0), palmitic acid (16:0), stearic acid (18:0), arachidic acid (20:0); the monounsaturated fatty acids, palmitoleic acid (16:1), oleic acid (18:1); the polyunsaturated fatty acids, linoleic acid (18:2), linolenic acid (18:3). Fourth instar nymphs had 5-10 times the quantities of fatty acids as compared to third instar nymphs and 1-3 times the quantities from adults. The fatty acid quantity differences between fourth and third instar nymphs were related to their size and weight differences. The percentage compositions for fatty acids from each lipid class were the same for the pooled groups of third and fourth instar nymphs. For nymphs and adults, triacylglycerols were the major source of fatty acids, with 18:1 and 16:0 acids as major components and the majority of the polyunsaturated fatty acids, 18:2 and 18:3 were present in the two phospholipid fractions, phosphatidylethanolamine and phosphatidylcholine. Evidence was obtained that whiteflies indeed synthesize linoleic acid and linolenic acid de novo: radiolabel from [2-(14)C] acetate was incorporated into 18:2 and 18:3 fatty acids of B. argentifolii adults and CGC-MS of pyrrolidide derivatives established double bonds in the Delta(9,12) and Delta(9,12,15) positions, respectively. PMID- 12761875 TI - Induction of glutathione S-transferases activities in Drosophila melanogaster exposed to phenol. AB - Studying the toxic effects of long-term exposing fruit flies to phenol is the object of this study. The induction of the glutathione S-transferases enzymatic activities, the change in the amount of mRNA related to phenol exposure, the change in survival rate of adult fruit flies, and the chemical interaction between phenol and benzene were the problems to be investigated. Glutathione S transferases were separated by affinity chromatography and the mRNAs levels were quantified by reverse-transcription polymerase chain reaction. Long-term feeding phenol to wild type fruit flies had caused some toxic effects included increasing the resistance to phenol toxicity, lowering the benzene toxicity, and induction of glutathione S-transferases enzymatic activities. But no significant change in the amount of glutathione S-transferases GstD1 and GstD5 mRNAs had occurred. From these results, we concluded that fruit flies could develop resistance to phenol by decreasing its toxicity; phenol was a inducer of glutathione S-transferases; phenol could increase the glutathione S-transferases enzymatic activities by increasing the amount of proteins; phenol exposure could decrease the benzene toxicity; no new glutathione S-transferase isozyme subunit was induced; and the level of GstD1 and GstD5 mRNAs did not significantly increase in phenol-treated strain. PMID- 12761877 TI - Identification, characterization, and expression of a novel alpha-tropomyosin isoform in cardiac tissues in developing chicken. AB - Tropomyosins are present in various muscle (skeletal, cardiac, and smooth) and non-muscle cells with different isoforms characteristic of specific cell types. We describe here a novel smooth/striated chimeric isoform that was expressed in developing chick heart in addition to the classically described TM-4 type. This novel alpha-Tm tropomyosin isoform, designated as alpha-Tm-2, contains exon 2a (in place of exon 2b). The known striated muscle isoform (alpha-Tm-1) was also expressed in embryonic hearts along with the striated muscle isoform of TM-4. In adult heart, TM-4 was expressed, however, expression of both alpha-Tm-1 and alpha Tm-2 isoforms was drastically reduced or downregulated. Interestingly, we were unable to detect the expression of alpha-Tm-2 in embryonic and adult skeletal muscle, however, the alpha-Tm-1 isoform is expressed in embryonic and adult skeletal muscle. Examination of other possible isoforms of the alpha-TM gene, i.e., alpha-smooth muscle tropomyosin (alpha-Sm), alpha-Fibroblast-1 (alpha-F1), and alpha-Fibroblast-2 (alpha-F2) revealed expression in embryonic hearts and a significant reduction of each of these isoforms in adult heart. In order to elucidate the role of the newly discovered tropomyosin isoform in chicken, we ectopically expressed the GFP fusion protein of alpha-Tm-1 and alpha-Tm-2 separately into cardiomyocytes isolated from neonatal rats. Each isoform was incorporated into organized myofibrils. Our results suggest that the alpha-TM gene may undergo both positive and negative transcriptional control in chicken hearts during development. PMID- 12761876 TI - Isolation and characterization of a novel venom protein from an endoparasitoid, Cotesia rubecula (Hym: Braconidae). AB - Insects are important vectors of diseases with remarkable immune defense capabilities. Hymenopteran endoparasitoids are adapted to overcome the host defense system and, therefore, are useful sources of immune-suppressing proteins. Not much is known about venom proteins in endoparasitoids, especially those that have a functional relationship with polydnaviruses (PDVs). Here, we describe the isolation and characterization of a small venom protein (Vn4.6) from an endoparasitoid, Cotesia rubecula, which interferes with the activation of the host hemolymph prophenoloxidase. The coding region for Vn4.6 is located upstream in the opposite direction of a gene coding for a C. rubecula PDV-protein (Crp32). PMID- 12761878 TI - Vitamin D enhances mitogenesis mediated by keratinocyte growth factor receptor in keratinocytes. AB - The hormonally active vitamin D metabolite, 1,25-dihydroxyvitamin D(3) (1,25(OH)(2)D(3)), and keratinocyte growth factor (KGF) belong to the network of autocrine and paracrine mediators in the skin. Both were shown to modulate keratinocyte proliferation, to reverse epidermal atrophy, to increase wound healing, and to reduce chemotherapy-induced alopecia. The overlap between their activities may suggest that vitamin D exerts some of its actions by modulation of KGF activities in the skin. This notion was examined by using HaCaT keratinocytes cultured in serum-free medium in the absence of exogenous growth factors and in the presence of the EGF receptor tyrosine kinase inhibitor AG 1478 that blocks their autonomous proliferation. These cells could be stimulated to proliferate by different fibroblast growth factors (FGFs). The relative mitogenic efficacy of basic FGF, acidic FGF, or KGF was in correlation with their affinities for the KGF receptor (KGFR). Forty-eight hour co-treatment with 1,25(OH)(2)D(3) enhanced KGFR-mediated cell proliferation in a dose dependent manner. Both ERK1/2 and c Jun N-terminal kinase (JNK) were activated by the FGFs. Treatment with 1,25(OH)(2)D(3) increased the activation of ERK but reduced the activation of JNK. Treatment with 1,25(OH)(2)D(3) increased the levels of KGFR in the presence but not in the absence of KGF, probably due to inhibition of ligand-induced receptor degradation. Inhibition of protein kinase C with bisindolylmaleimide did not interfere with the effect of 1,25(OH)(2)D(3) on KGFR-mediated ERK activation. Our results support the notion that the paracrine KGF-KGFR system in the skin can act in concert with the autocrine vitamin D system in keratinocytes to promote keratinocyte proliferation and survival under situations of stress and injury. PMID- 12761879 TI - Increased gap junctional intercellular communication is directly related to the anti-tumor effect of all-trans-retinoic acid plus tamoxifen in a human mammary cancer cell line. AB - Additive effects against tumor cells might be achieved by combining anti neoplastic agents directed against one or more altered mechanisms in cancer. We investigated the participation of gap junctional intercellular communication (GJIC), which is commonly dysfunctional in tumor cells as a possible mediating mechanism of the effect of all-trans-retinoic acid (RA) and tamoxifen (Tx) in MCF 7 human breast cancer cell lines. The combination of RA + Tx stimulated GJIC in approximately 53 +/- 3% of MCF-7 cells as early as after 6 h of treatment remaining communicated through 144 h of culture. The GJIC enhancement occurred along with immunolocalization of Cx26 and 43 at the membrane of contacting cells and correlated with higher protein levels. Cx40 immunoreactive plaques were detected at cell-to-cell contacts during 48 h of RA + Tx treatment that did not involve higher protein expression, to the contrary, a downregulation occurred after 72 h of treatment. Cell proliferation inhibition upon RA + Tx exposure was observed with optimal effects at 96-120 h of culture with an accumulation of cells primarily in G2/M and G0/G1 cell cycle boundaries. An enhancement of the pre-existing E-cadherin levels was observed after drug exposure along with a downregulation of Bcl-2 and C-myc protein levels and a reduction of telomerase activity, suggesting partial tumor phenotype reversion. Blockage of the RA + Tx induced GJIC with 18-beta-glycyrrhetinic acid (beta-Gly) prevented in 34% the inhibition of MCF-7 proliferation and the E-cadherin increment in 30% at 96 h of culture. GJIC blockage did not alter the downregulation of Bcl-2, c-Myc, or telomerase activity induced by RA + Tx. Our results showed the participation of GJIC as a mediator mechanism of the combined action of RA and Tx in MCF-7 cells. The chemopreventive modulation of GJIC might represent an approachable alternative for the improvement of cancer therapy. PMID- 12761880 TI - Expression of CXCR4 and CXCL12 (SDF-1) in human prostate cancers (PCa) in vivo. AB - Human prostate cancers (PCa) express great variability in their ability to metastasize to bone. The identification of molecules associated with aggressive phenotypes will help to define PCa subsets and will ultimately lead to better treatment strategies. The chemokine stromal-derived factor-1 (SDF-1 or CXCL12) and its receptor CXCR4 are now known to modulate the migration and survival of an increasing array of normal and malignant cell types including breast, pancreatic cancers, glioblastomas, and others. The present investigation extends our previous investigations by determining the expression of CXCR4 and CXCL12 in humans using high-density tissue microarrays constructed from clinical samples obtained from a cohort of over 600 patients. These data demonstrate that CXCR4 protein expression is significantly elevated in localized and metastastic cancers. At the RNA level, human PCa tumors also express CXCR4 and message, but overall, they were not significantly different suggesting post-transcriptional regulation of the receptor plays a major role in regulating protein expression. Similar observations were made for CXCL12 message, but in this case more CXCL12 message was expressed by metastastic lesions as compared to normal tissues. PCa cell lines also express CXCL12 mRNA, and regulate mRNA expression in response to CXCL12 and secrete biologically active protein. Furthermore, neutralizing antibody to CXCL12 decreased the proliferation of bone homing LNCaP C4-2B and PC3 metastastic tumor cells. These investigations provide important new information pertaining to the molecular basis of how tumors may 'home' to bone, and the mechanisms that may account for their growth in selected end organs. PMID- 12761882 TI - Overexpression of cyclin D1 inhibits TNF-induced growth arrest. AB - Although activated macrophages destroy cancer cells more effectively than normal cells, the facility to escape activated macrophages is a characteristic of tumor cells. One of the mechanisms responsible for the specific killing of tumor cells by macrophages is the production of the cytokine tumor necrosis factor alpha (TNF). Therefore, resistance to TNF may provide such cancer cells a selective advantage against host elimination. In the present work we explore the possibility that cyclin D1 overrides the cytostatic effect of TNF. We show that TNF induces p21(waf1) protein in malignant melanoma A375 cells and its binding to CDK2/4 and 6 proteins, and thereby inhibiting the activity of these complexes. This inhibition leads the cells to a G1 arrest. Overexpression of cyclin D1 in these cells makes them insensitive to TNF treatment with the recovery of CDK activity, however, is unable to overcome the inhibitory action of etoposide blocking the cells on G2/M. The bypass of TNF-induced G1 arrest seems to be related to the increase in the stability of cyclin D bound CDK complexes, increasing the total amount of CDK2/4 and 6 complexes and leading to a functional down titration of the p21(waf1) molecules. In these conditions the TNF-induced increase of p21(waf1) is not sufficient to inhibit the high amount of cyclin D bound complexes. This hypothesis is supported by the fact that a reduction in the levels of p21(waf1) protein, induced by the expression of a mRNA antisense against p21(waf1), is also able to bypass of TNF-induced arrest. Our results confirm that p21(waf1) has an essential role in TNF-induced arrest and that the deregulation of cyclin D1 may be one of the mechanisms to escape physiological signals to restrict tumoral growth. PMID- 12761881 TI - TGF-beta1-induced Smad 3 binding to the Smad 7 gene: knockout of Smad 7 gene transcription by sense phosphorothioate oligos, autoregulation, and effect on TGF beta1 secretion: bleomycin acts through TGF-beta1. AB - Bleomycin produces its fibrogenic effect, at least in part, by TGF-beta1 secretion. Treatment of IMR-90 human embryonic lung fibroblasts with bleomycin at 0.5 microg/ml results in a 1.6-fold increase of TGF-beta1 as determined by a specific ELISA assay for TGF-beta1 after acidification of the conditioned media. This elevation of TGF-beta1 secretion is furthermore enhanced in vivo by TGF beta1 autoinduction of the TGF-beta1 gene. To demonstrate TGF-beta1 autoinduction, the fibroblasts were pretreated with 12.5 ng/ml TGF-beta1, washed extensively to remove any residual TGF-beta1, and then allowed to incubate for 24 h in AIM V synthetic serum-free media. The media when assayed using the ELISA assay contained a 1.6-fold increase of TGF-beta1. The distal promoter of the human TGF-beta1 gene contains a Smad 3 element (CAGGACA), which is homologous to the Smad 3 binding element motif (CAGA). The nuclear extracts of human embryonic lung fibroblasts treated for either 15 min or 24 h with TGF-beta1 did not demonstrate specificity of binding of a protein(s) to the homologous Smad 3 element as determined by cold wild-type oligodeoxynucleotide competition experiments. However, specific Smad 3 binding to the Smad 3 element (GTCTAGAC) found in proximal promoter of the Smad 7 gene was observed by cold oligo competition and supershift assays using a goat polyclonal Smad 3 antibody in the presence and absence of an N-terminal Smad 3 peptide. To determine the functionality of this Smad 3 binding to the Smad 3 element in the proximal promoter of the Smad 7 inhibitory gene to TGF-beta1 secretion, fibroblasts were transiently pretransfected with double-stranded phosphorothioate oligo "decoys" containing the Smad 7/Smad 3 element in the presence of plasmin to convert latent TGF-beta1 to active TGF-beta1. Under these conditions, which simulate the in vivo situation of 2.2-fold increase of total active TGF-beta1 was observed. Fibroblasts were also pretransfected with these double-stranded oligo "decoys," washed, then treated with TGF-beta1, washed and incubated in AIM V for an additional 24 h. In this latter experiment, a superinduction of TGF-beta1 secretion was observed. We propose that these oligo "decoys" bind Smad 3 preventing this initiation factor from binding to the Smad 7/Smad 3 element thereby decreasing the transcription of the Smad 7 gene. The decrease of the inhibitory Smad 7 would result in less binding of this Smad inhibitor to the Type I TGF-beta receptor and less antagonism of active TGF-beta1, more autoinduction of the TGF-beta1 gene, and more of the fibrogenic effects of TGF-beta1. PMID- 12761883 TI - Thrombospondin-1-induced vascular smooth muscle cell chemotaxis: the role of the type 3 repeat and carboxyl terminal domains. AB - Thrombospondin-1 (TSP-1), an acute phase reactant implicated in vascular disease, is a matricellular glycoprotein with six domains that confer different functions. The authors have shown TSP-1 induces vascular smooth muscle cell (VSMC) chemotaxis via extracellular signal-regulated kinases-1 and -2 (ERK) and p38 kinase (p38) and that a fusion protein of the carboxyl terminal (COOH) and type 3 repeat (T3) domains independently induce VSMC chemotaxis. The purpose of this study was to determine whether COOH-, T3-induced VSMC chemotaxis, or both, is dependent upon ERK or p38 activation. To determine if the T3, COOH, or type 2 repeat domain (T2, control domain not associated with chemotaxis) activate ERK, p38, or both, VSMCs were exposed to each fusion protein (20 microg/ml for 15, 30, 60, or 120 min), serum-free media (SFM, negative control), or TSP-1 (20 microg/ml for 30 min, positive control). Western immunoblotting was performed for activation studies. Using a microchemotaxis chamber, VSMCs pre-incubated in SFM, DMSO (vehicle control), PD98059 (10 microM), or SB202190 (10 microM) were exposed to each domain, TSP-1, or SFM. After 4 h (37 degrees C), migrated VSMCs were recorded as cells/five fields (400 x) and analyzed by paired t-test. ERK was activated by T2, T3, and COOH. However, p38 was activated by T3 and COOH, but not T2. T3 and COOH-induced VSMC chemotaxis were inhibited by PD98059 or SB202190, but more completely by SB202190. The T2 domain had no effect on VSMC chemotaxis. These results suggest activation of the p38 pathway may be more specific than ERK for COOH- and T3-induced VSMC chemotaxis. PMID- 12761884 TI - The sphingosine 1-phosphate receptor S1P4 regulates cell shape and motility via coupling to Gi and G12/13. AB - Sphingosine 1-phosphate (S1P) receptors represent a novel subfamily of G-protein coupled receptors binding S1P specifically and with high affinity. Although their in vivo functions remain largely unknown, in vitro extracellular application of S1P induces distinct S1P receptor-dependent cellular responses including proliferation, differentiation, and migration. We have analyzed signaling pathways engaged by S1P(4), which is highly expressed in the lymphoid system. Here we show that S1P(4) couples directly to Galpha(i) and even more effectively to Galpha(12/13)-subunits of trimeric G-proteins, but not to Galpha(q) unlike other S1P receptors. Consequently, CHO-K1 cells ectopically expressing S1P(4) potently activate the small GTPase Rho and undergo cytoskeletal rearrangements, inducing peripheral stress fiber formation and cell rounding, upon S1P stimulation. Overexpression of S1P(4) in Jurkat T cells induces pertussis toxin sensitive cell motility even in the absence of exogenously added S1P. In addition, S1P(4) is internalized upon binding of S1P. The capacity of S1P(4) to mediate cellular responses, such as motility and shape change through Galpha(i)- and Galpha(12/13)-coupled signaling pathways may be important for its in vivo function which is currently under investigation. PMID- 12761885 TI - Phospholipase C-protein kinase C mediated phospholipase D activation pathway is involved in tamoxifen induced apoptosis. AB - Tamoxifen (TAM) is the endocrine therapeutic agent the most widely used in the treatment of breast cancer, and it operates primarily through the induction of apoptosis. In this study, we attempted to elucidate the non-ER mediated mechanism behind TAM treatment, involving the phospholipase C-protein kinase C (PLC-PKC) mediated phospholipase D (PLD) activation pathway, using multimodality methods. In TAM treated MCF7 cells, the PLC and PLD protein and mRNA levels increased. Phosphatidylethanol (PEt) and diacylglycerol (DAG) generation also increased, showing increased activity of PLD and PLCgamma1. Translocation of PKCalpha, from cytosol to membrane, was observed in TAM treated cells. By showing that both PKC and PLC inhibitors could reduce the effects of TAM-induced PLD activation, we confirmed the role of PKC and PLC as upstream regulators of PLD. Finally, we demonstrated that TAM treatment reduced the viability of MCF7 cells and brought about rapid cell death. From these results, we confirmed the hypothesis that TAM induces apoptosis in breast cancer cells, and that the signal transduction pathway, involving PLD, PLC, and PKC, constitutes one of the possible mechanisms underlying the non-ER mediated effects associated with TAM. PMID- 12761886 TI - Flavonoids inhibit VEGF/bFGF-induced angiogenesis in vitro by inhibiting the matrix-degrading proteases. AB - Flavonoids have been proposed to act as chemopreventive agents in numerous epidemiological studies and have been shown to inhibit angiogenesis and proliferation of tumor cells and endothelial cells in vitro. Angiogenesis requires tightly controlled extracellular matrix degradation mediated by extracellular proteolytic enzymes including matrix metalloproteinases (MMPs) and serine proteases, in particular, the urokinase-type plasminogen activator (uPA) plasmin system. In this study, we have investigated the antiangiogenic mechanism of the flavonoids, genistein, apigenin, and 3-hydroxyflavone in a human umbilical vein endothelial cell (HUVEC) model. The stimulation of serum-starved HUVECs with vascular endothelial growth factor/basic fibroblast growth factor (VEGF/bFGF) caused marked increase in MMP-1 production and induced the pro-MMP-2 activation accompanied by the increase in MT1-MMP expression. However, pretreatment with flavonoids before VEGF/bFGF stimulation completely abolished the VEGF/bFGF stimulated increase in MMP-1 and MT1-MMP expression and pro-MMP-2 activation. Genistein blocked VEGF/bFGF-stimulated increase in TIMP-1 expression and decrease in TIMP-2 expression. Apigenin and 3-hydroxyflavone further decreased TIMP-1 expression below basal level and completely abolished TIMP-2 expression. VEGF and bFGF stimulation also significantly induced uPA expression, most strikingly the level of 33 kDa uPA, and increased the expression of PA inhibitor (PAI)-1. Genistein, apigenin, and 3-hydroxyflavone effectively blocked the generation of 33 kDa uPA, and further decreased the activity of the 55 kDa uPA and the expression of PAI-1 below the basal level. In conclusion, these data suggest that genistein, apigenin, and 3-hydroxyflavone inhibit in vitro angiogenesis, in part via preventing VEGF/bFGF-induced MMP-1 and uPA expression and the activation of pro-MMP-2, and via modulating their inhibitors, TIMP-1 and -2, and PAI-1. PMID- 12761887 TI - Regulation of p27 in the process of neuroblastoma N2A differentiation. AB - Neuronal differentiation implies morphological and biochemical changes to generate a specialized neuron. N2A neuroblastoma cells can be promoted to undergo differentiation associated to neurites outgrowth, a process linked to the arrest of cell division. Using N2A cells as a model, we investigated the detailed molecular aspects on the involvement of p27 in dibutyryl cAMP-induced neuronal differentiation. In the undifferentiated N2A phenotype, an unusually high level of accumulated p27 protein mass was evidenced. Data suggest that in proliferating cells, p27 could be sequestered by direct interaction with cyclin D1, thus preventing its inhibitory action on cell cycle Cdks. Studies also indicate that p27 is functionally active and that its loss of action on Cdks in proliferating cells is due to its strong association with cyclin D1. Therefore, when cell differentiation is triggered, the action of p27 on Cdks seems to depend on both p27 and cyclin D1 degradation during the early steps of differentiation followed by late events of re-synthesis of active p27. In this context, an overexpression of p27 after N2A transfection with a mouse p27 clone induces the outgrowth of neurites associated with a decrease in cyclin D1 expression. On the other hand, treatment of N2A undifferentiated cells with c-myc antisense oligonucleotides led to a decrease in p27 and cyclin D1 levels, similar events as those in early stages of cell differentiation. Studies suggest that blockage in c-myc expression triggers early events in neuronal differentiation. These studies are of the utmost importance to elucidate regulatory mechanisms of molecules that play a critical role in the transition from a proliferating phenotype to differentiated cells. PMID- 12761888 TI - Expression of genes for certain enzymes of pyrimidine and purine salvage pathway in peripheral blood leukocytes collected from patients with Graves' or Hashimoto's disease. AB - Increased activities of some enzymes, which participate in pyrimidine and purine salvage pathway, were found in blood fractions of patients suffering from different autoimmunological diseases, thyroid diseases included. The aim of the study was to estimate the expression of genes, specific for deoxycytidine kinase (dCK, EC 3.7.1.74), thymidine kinase 1 (TK1; EC 2.7.1.21), and adenosine deaminase (ADA, EC 3.5.4.4) in blood leukocytes, collected from patients with autoimmunological thyroid diseases (AITD), i.e., Graves' or Hashimoto's disease. The total mRNA was isolated from peripheral blood leukocytes and, afterwards, submitted to reverse transcription (RT), with the following amplification of genes encoding for particular examined enzymes and beta-actin, as a supervisory gene [RT-polymerase chain reaction (RT-PCR)]; ADA gene was amplified with the use of three different primer pairs (ADA3, ADA4, and ADA5). PCR products were electrophoresed in 8% polyacrylamide gel and then, submitted to densitometric analysis. The levels of expression of all the examined genes in leukocytes from patients with either Graves' or Hashimoto's disease were significantly increased when compared to those in controls; above a twofold elevation of expression of TK1, ADA4, and ADA5 genes was observed. In conclusion, the changes of activities of salvage enzymes in patients with AITD occur likely at transcription level; the measurement of gene expression for purine and pyrimidyne salvage enzymes may likely help explain the mechanism of autoimmune diseases, being also significant in the diagnostics and/or monitoring of AITD. PMID- 12761889 TI - ATP modulates load-inducible IL-1beta, COX 2, and MMP-3 gene expression in human tendon cells. AB - Tendon cells receive mechanical signals from the load bearing matrices. The response to mechanical stimulation is crucial for tendon function. However, overloading tendon cells may deteriorate extracellular matrix integrity by activating intrinsic factors such as matrix metalloproteinases (MMPs) that trigger matrix destruction. We hypothesized that mechanical loading might induce interleukin-1beta (IL-1beta) in tendon cells, which can induce MMPs, and that extracellular ATP might inhibit the load-inducible gene expression. Human tendon cells isolated from flexor digitorum profundus tendons (FDPs) of four patients were made quiescent and treated with ATP (10 or 100 microM) for 5 min, then stretched equibiaxially (1 Hz, 3.5% elongation) for 2 h followed by an 18-h-rest period. Stretching induced IL-1beta, cyclooxygenase 2 (COX 2), and MMP-3 genes but not MMP-1. ATP reduced the load-inducible gene expression but had no effect alone. A medium change caused tendon cells to secrete ATP into the medium, as did exogenous UTP. The data demonstrate that mechanical loading induces ATP release in tendon cells and stimulates expression of IL-1beta, COX 2, and MMP-3. Load induced endogenous IL-1beta may trigger matrix remodeling or a more destructive pathway(s) involving IL-1beta, COX 2, and MMP-3. Concomitant autocrine and paracrine release of ATP may serve as a negative feedback mechanism to limit activation of such an injurious pathway. Attenuation or failure of this negative feedback mechanism may result in the progression to tendinosis. PMID- 12761890 TI - Identification of a domain within MDMX-S that is responsible for its high affinity interaction with p53 and high-level expression in mammalian cells. AB - The MDMX gene product is related to the MDM2 oncoprotein, both of which interact with the p53 tumor suppressor. A novel transcript of the MDMX gene has been previously identified that has a short internal deletion of 68 base pairs, producing a shift in the reading frame after codon 114, resulting in the inclusion of 13 novel amino acids (after residue 114) followed by a stop codon at amino acid residue 127. This truncated MDMX protein, termed MDMX-S, represents only the p53 binding domain and binds and inactivates p53 better than full-length MDMX or MDM2. Here we show that when expressed in cells, MDMX-S is targeted more efficiently to the nucleus than MDMX. MDMX-S suppresses p53-mediated transcription from a p53 target promoter better than full-length MDMX. The DNA damage inducibility of these p53 responsive promoters was suppressed better by MDMX-S than by MDMX. Analysis of the MDMX-S protein indicated that the 13 novel amino acids at its carboxy terminus was responsible for high affinity binding to p53 in vitro and for high level expression of the protein in cells. Deletion of this 13 amino acid sequence resulted in a protein that was not able to bind p53 and was not able to be expressed well in cells. Taken together, these data point to an important domain within MDMX-S that enables it to function well in vivo to block p53 activity. Published 2003 Wiley-Liss, Inc. PMID- 12761891 TI - hMad4, c-Myc endogenous inhibitor, induces a replicative senescence-like state when overexpressed in human fibroblasts. AB - Mad family proteins have an antagonistic action on Myc-dependent cell proliferation and transformation. We isolated a human cDNA clone, human Mad4 (hMad4), encoding a polypeptide of 209 amino acid residues, exhibiting 90% identity with mouse Mad4. Northern blot analysis shows that hMad4 probe hybridizes to a 3.8 kb message; its expression is highest in quiescent human WI38 fibroblasts. Among tissues, hMad4 mRNA is most abundant in brain, lung, and muscle. Consistent with other members of the Mad family, hMad4 can repress the transactivation activity of Myc/Max heterodimers on an E-box chloramphenicol acteyl transferase (CAT) reporter plasmid; inhibition of both proliferation and clonogenic formation of hMad4-infected cells correlates with the in vitro reporter repression. Moreover, infection of young human fibroblasts induces a replicative senescence-like state. This phenotype was accompanied by s-beta galactosidase and PAI-1 expression. These results suggest that hMad4 might be an important regulator of replicative senescence in human cells. PMID- 12761892 TI - Retaining of the assembly capability of vimentin phosphorylated by mitogen activated protein kinase-activated protein kinase-2. AB - Intermediate filament (IF) networks can be regulated by phosphorylation of unit proteins, such as vimentin, by specific kinases leading to reorganization of the IF filamentous structure. Recently, we identified mitogen-activated protein kinase-activated protein kinase-2 (MAPKAP kinase-2) as a vimentin kinase (Cheng and Lai [1998] J. Cell. Biochem. 71:169-181). Herein we describe the results of further in vitro studies investigating the effects of MAPKAP kinase-2 phosphorylation on vimentin and the effects of the phosphorylation on the filamentous structure. We show that MAPKAP kinase-2 mainly phosphorylates vimentin at Ser-38, Ser-50, Ser-55, and Ser-82, residues all located in the head domain of the protein. Surprisingly, and in stark contrast to phosphorylation by most other kinases, phosphorylation of vimentin by MAPKAP kinase-2 has no discernable effect on its assembly. It suggested that structure disassembly is not the only obligated consequence of phosphorylated vimentin as regulated by other kinases. Finally, a mutational analysis of each of the phosphorylated serine residues in vimentin suggested that no single serine site was primarily responsible for structure maintenance, implying that the retention of filamentous structure may be the result of the coordinated action of several phosphorylated serine sites. This also shed new lights on the functional task(s) of vimentin that is intermediate filament proteins might provide a phosphate reservoir to accommodate the phosphate surge without any structural changes. PMID- 12761893 TI - DcR3/TR6 modulates immune cell interactions. AB - DcR3/TR6, a secreted protein, is a member of TNF receptor family. Its ligands include FasL, LIGHT, and TL1A, all TNF family members. TR6 can interfere with FasL- or LTbetaR-mediated apoptosis; it can also inhibit T-cell costimulation by blocking the two-way signaling between TR2 and LIGHT, and the one-way signaling from TL1A to DR3. In this study, we discovered that TR6 was secreted by peripheral blood mononuclear cells (PBMC) stimulated by T-cell mitogens. It inhibited actin polymerization of T cells upon mitogen stimulation, and repress T cell pseudopodium formation, which is known to be important for cell-cell interaction. As a consequence, T-cell aggregation stimulated by alloantigens, anti-CD3 or PHA was suppressed by either soluble or solid phase TR6-Fc. This result suggests that TR6 might regulate T-cell interaction with other cells such as antigen-presenting cells (APC) or their fellow T cells by preventing them from forming inseparable cell clusters, which are undesirable for the progression of immune responses. PMID- 12761894 TI - Nitric oxide synthase I mediates osteoclast activity in vitro and in vivo. AB - Bone resorption is responsible for the morbidity associated with a number of inflammatory diseases such as rheumatoid arthritis, orthopedic implant osteolysis, periodontitis and aural cholesteatoma. Previous studies have established nitric oxide (NO) as a potentially important mediator of bone resorption. NO is a unique intercellular and intracellular signaling molecule involved in many physiologic and pathologic pathways. NO is generated from L arginine by the enzyme nitric oxide synthase (NOS). There are three known isoforms of NOS with distinct cellular distributions. In this study, we have used mice with targeted deletions in each of these isoforms to establish a role for these enzymes in the regulation of bone resorption in vivo and in vitro. In a murine model of particle induced osteolysis, NOS I-/- mice demonstrated a significantly reduced osteoclast response. In vitro, osteoclasts derived from NOS I-/- mice were larger than wild type controls but demonstrated decreased resorption. Although NOS I has been demonstrated in osteoblasts and osteocytes as a mediator of adaptive bone remodeling, it has not previously been identified in osteoclasts. These results demonstrate a critical role for NOS I in inflammatory bone resorption and osteoclast function in vitro. PMID- 12761895 TI - Apoptotic effect of cleaved high molecular weight kininogen is regulated by extracellular matrix proteins. AB - We previously reported that cleaved high molecular weight kininogen (HKa) and its domain 5 (D5) inhibit critical steps required for angiogenesis and in vivo neovascularization (Colman et al. 2000: Blood 95:543-550). We have further shown that D5 is able to induce apoptosis of endothelial cells, which may represent a critical part of the anti-angiogenic activity of HKa and D5 (Guo et al. 2001: Arterioscler Thromb Vasc Biol 21:1427-1433). In this study, we demonstrate that HKa- and D5-induced apoptosis is closely correlated with their anti-adhesive effect. An important new finding is that the apoptotic activity of HKa and D5 is highly regulated by their interactions with different extracellular matrix (ECM) proteins. HKa inhibited cell adhesion to vitronectin (Vn, 90%) and gelatin (Gel) (40%), but it had no apparent effect on cell adhesion to fibronectin (Fn). D5 showed a similar pattern on cell adhesion but was less potent than HKa. HKa induced apoptosis of endothelial cells grown on Vn and Gel but not cells grown on Fn which closely parallels with its anti-adhesive potency. Further results revealed that the anti-adhesive effect and the apoptotic effect of HKa are associated with its ability to inhibit phosphorylation of focal adhesion kinase (FAK) and paxillin, two important signal molecules required for cell adhesion and cell viability. We conclude that the anti-adhesive activity of HKa and D5 is responsible for their apoptotic effect and that Vn is likely an ECM component that mediates the effect of HKa and D5. PMID- 12761896 TI - Phytoestrogen genistein acts as an estrogen agonist on human osteoblastic cells through estrogen receptors alpha and beta. AB - Genistein, a soybean isoflavone, has estrogen-like activity in mammals, including the prevention of bone loss. However, whether its mechanism of action on bone turnover is distinct from that of estrogen or raloxifene is unknown. Although genistein has been reported to bind both estrogen receptor (ER) isoforms (alpha and beta), little is known concerning differential activation of gene expression via these ER isoforms. To examine this question, comparison of the responses of normal fetal osteoblast (hFOB) cells stably expressing either ERalpha (hFOB/ERalpha9) or ERbeta (hFOB/ERbeta6), to treatment with genistein, 17beta estradiol (E(2)) or raloxifene were conducted. In hFOB/ERalpha9 cells, both genistein and E(2) increased the endogenous gene expression of the progesterone receptor (PR), the proteoglycan versican, and alkaline phosphatase (AP), but inhibited osteopontin (OP) gene expression and interleukin-6 (IL-6) protein levels. Raloxifene had no effect on these bone markers. Genistein, but not raloxifene, also mimicked E(2) action in the hFOB/ERbeta6 cells increasing PR gene expression and inhibiting IL-6 production. To determine whether the gene regulatory actions of genistein in human osteoblast cells occur at the level of transcription, its action on the transcriptional activity of a PR-A promoter reporter construct was assessed. Both genistein and E(2) were found to stimulate the PR promoter in the hFOB cell line when transiently co-transfected with either ERalpha or ERbeta. Whereas hFOB cell proliferation was unaffected by E(2), raloxifene or genistein at low concentrations, higher concentrations of genistein, displayed significant inhibition. Together, these findings demonstrate that genistein behaves as a weak E(2) agonist in osteoblasts and can utilize both ERalpha and ERbeta. PMID- 12761897 TI - [The orientation and its significance of water channel protein in the mouse inner ear]. AB - OBJECTIVE: To examine the distribution of various subtypes of water channel protein (aquaporin, Aqp) in various structures of the mouse inner ear. METHODS: Thirty mice with white color were used in this study and were cardiacally perfused. The temporal bones were taken out and were processed and sectioned by paraffin-embedded technique. The sections were labeled with fluorescent antibody by immunohistochemical method. The distribution of Aqp1, 3, 4, 5, 7, 9 were confirmed in the inner ear of mouse. RESULTS: The constant and clear fluorescent reaction could be observed in the inner ear tissue with the first antibody concentration of 1:200 for Aqp-1 and 1:100 for Aqp-3, 7, 9. But the reaction of Aqp-4 and 5 could not be found with the concentration of 1:50, even 1:30. Aqp-1 was labeled in the round window membrane, spiral ligament, endolymphatic sac and duct, utricle and saccule and the wall of inner ear blood vessel; Aqp-3 in the spiral limb, vestibular lip, internal and external sulci, basilar membrane and basilar membrane crest, endolymphatic sac and duct, membranous semicircular canal and utricle and saccular macula; Aqp-7 in the stria vascularis, basilar membrane, Reissner's membrane, utricle and saccule and their maculae; Aqp-9 in the spiral limb, vestibular lip internal and external sulci, Reissner's membrane, membranous semicircular canal, saccule and its macula. CONCLUSION: Aqp-1, 3, 7, 9 were extensively distributed in various tissues of mouse inner ear. Their distribution sites and reaction degree were different, and mainly located in the structures related to the endolymph. PMID- 12761899 TI - [Preliminary study of the facial nerve regeneration in the chamber: the influence of myelin basic protein]. AB - OBJECTIVE: To study the role of exogenous myelin basic protein(MBP) in neural repairment. METHODS: Adult New Zealand rabbits were employed in vivo preparation. A 12 microL nerve growth chamber was created by suturing the proximal and distal stumps of a transected facial never (FN) trunk into a tube. The regenerated nerves within the chambers were dissected and fixed for histological studies with light microscope at 4, 6 and 8 weeks respectively following the surgery. RESULTS: Morphological analysis of nerves showed no difference between the MBP and control group in the size of the regeneration FN within the chambers, diameters of myelinated axons, thickness of myelin sheath and number of myelin axons grew into the distal end of chamber at 4 weeks. At 6 and 8 weeks after operation, the MBP group showed a more mature-appearance regenerative nerve comparing to control group. Especially, the enhancement of maturation in the regeneration axons was very noticeable at 6 weeks. CONCLUSION: The study showed that pharmacological administration of exogenous MBP within a chamber at the time of entubational nerve repair enhances regeneration of myelinated axons across the sectioned ends of FN. PMID- 12761898 TI - [Expression of bcl-2 in facial motoneurons and its ultrastructural localization following facial nerve injury]. AB - OBJECTIVE: To explore the expression of bcl-2 in facial motoneurons and its subcellular distribution. METHODS: Wistar rats were used in this study. Facial nerve transection was performed at stylomastoid foramen or internal acoustic meatus. Facial nerve crush was made at stylomastiod foramen. The animal survived for 1, 3, 7, 15, 30 and 60 days respectively. Facial nucleus was treated with bcl 2 monoclonal antibody or bcl-2 DIG-labelling probe and studied with immunohistochemistry and in situ hybridization. The bcl-2 positive motoneuron was investigated with immuno-electron microscope. RESULTS: It was demonstrated that bcl-2 protein level was corresponded with bcl-2 mRNA expression. The level of bcl 2 expression in facial motoneurons was high in normal facial nerve. It increased on the first day and declined on the third day post-transection in facial motoneuron. It reached the lowest level on the 15th days following facial nerve injury (P < 0.05). The expression recovered to normal level in two months (P > 0.05). After facial nerve transected, the reduction of bcl-2 expression was more significant when facial nerve transected close to facial nucleus than that far from facial nucleus (P < 0.05). Comparing to facial nerve transection in stylomastoid foramen, there was more intensive bcl-2 expression following facial nerve crush (P < 0.05). Further study showed that bcl-2 primarily resided in the nuclear envelop, endoplasmic reticulum and mitochondrial membrane. CONCLUSIONS: These data indicated that high level bcl-2 protein may prevent facial motoneuron death following facial nerve injury. It is suggested that overexpression bcl-2 by transgene may prevent facial motoneurons death. PMID- 12761900 TI - [Effects of acute infrasound exposure on vestibular and auditory functions and the ultrastructural changes of inner ear in the guinea pig]. AB - OBJECTIVE: To define the effects of acute infrasound exposure on vestibular and auditory functions and the ultrastructural changes of inner ear in guinea pigs. METHODS: The animals involved in the study were exposed to 8 Hz infrasound at 135dB SPL for 90 minutes in a reverberant chamber. The sinusoidal pendular test (SPT), auditory brainstem response (ABR) and distortion product otoacoustic emissions (DPOAE) were respectively detected pre-exposure and at 0(within 2 hrs), 2 and 5 day after exposure. The ultrastructures of the inner ear were observed by scanning electron microscopy. RESULTS: The slow-phase velocity and the frequency of the vestibular nystagmus elicited by sinusoidal pendular test (SPT) declined slightly following infrasound exposure, but the changes were not significant (P > 0.05). No differences in the ABR thresholds, the latencies and the interval peak latencies of I, III, V waves were found between the normal and the experimental groups, and among experimental groups. The amplitudes of DPOAE at any frequency declined remarkably in all experimental groups. The ultrastructures of the inner ear were damaged to different extent. CONCLUSION: Infrasound could transiently depress the excitability of the vestibular end-organs, decrease the function of OHC in the organ of Corti and cause damage to the inner ear of guinea pigs. PMID- 12761901 TI - [Fabrication of cartilage in predetermined shapes from human nasoseptal chondrocytes with tissue engineering method]. AB - OBJECTIVE: To investigate the feasibility of fabricating a new cartilage in predetermined shapes with tissue engineering methods. METHODS: Human nasoseptal chondrocytes were seeded onto a nonwoven mesh of polyglycolic acid(PGA) to form a cell-PGA construct. The construction was then configured in sheet and tube shapes, and implanted subcutaneously into the dorsa of 11 athymic mice. The specimens were harvested 4, 6, 8 weeks after implantation and subjected to gross morphologic and histologic analysis. RESULTS: Gross observation showed that the predetermined sheet and tube shapes of new cartilage were formed. Histological observation demonstrated that new mature cartilages were formed in 6-week. A Masson's trichrome stain showed the interwining bands of collagen at the periphery of the cartilage. Staining of Safranin O evaluated that the new cartilage was bound of glycosaminoglycan. In the control group, the PGA of the specimens were completely absorbed at 6 weeks. CONCLUSION: Human nasoseptal chondrocytes-PGA construct could develop into a new cartilage in predetermined shapes in athymic mice. PMID- 12761903 TI - [Diagnosis and treatment of horizontal canal benign paroxysmal positional vertigo]. AB - OBJECTIVE: To explore effective methods for the diagnosis and treatment of horizontal-canal benign paroxysmal positional vertigo (HC-BPPV). METHODS: Medical records from nine patients with HC-BPPV, treated between July 1996 and March 2000, were retrospectively analyzed. Data of complete history, audiograms, positional tests and neuro-otological examinations were collected. All patients were treated with a particle repositioning maneuver called the "barbecue rotation" which starts with the patient in the supine position and consists of three 90-degree head rotations towards the unaffected ear. RESULTS: HC-BPPV was characterized by brief attacks of intense vertigo that were induced mainly by rolling over in bed (9/9) and turning the head to either side while upright (5/9). In most cases, rotation to the pathological side from supine position caused a very intense horizontal nystagmus beating towards the undermost ear. Findings such as latency and fatigability, which are common features of posterior canal BPPV (PC-BPPV), were not present. After the barbecue rotation, all patients had immediate and sustained relief of their attacks during the 4 to 15 months' follow up. CONCLUSION: HC-BPPV is different from PC-BPPV and other vertiginous diseases in typical presentations and positional testing results. The barbecue rotation is a successful method for curing the disorder. PMID- 12761902 TI - [An association study of histocompatibility leukocyte antigen-class II with Meniere's disease]. AB - OBJECTIVE: To investigate the relation between histocompatibility leukocyte antigen (HLA)-class II DRB1 and Meniere's disease in Chinese. METHODS: Polymerase chain reaction-sequence specific primers (PCR-SSP) technique was used. HLA-class II allele distribution were measured in 60 patients with Meniere's disease, in comparison with those in 85 normal population. RESULTS: The results showed that the frequency of DRB1*09 allele was significantly lower in Meniere's disease patients than in those of controls (relative risk = 0.17, P < 0.01). CONCLUSION: HLA-DRB1*09 of the patients with Meniere's disease was significant low. It implied that DRB1*09 may be a protective gene for Meniere's disease. PMID- 12761904 TI - [Analysis of the clinical salience for sudden hearing loss in acoustic neuroma]. AB - OBJECTIVE: To avoid misdiagnosis and misapplied therapy of acoustic neuroma. METHODS: 92 cases of acoustic neuroma treated between 1983 and 1997 were retrospectively reviewed. Among them, 20 patients (21 ears, 21.7%) presented with sudden hearing loss as the starting symptoms. The diagnosis was based on audiological and radiologic examinations. RESULTS: The clinical and audiological analysis demonstrated that 57.1% of the ears showed hearing loss over 71 dBHL, and all ears had deteriorated ABRs. Acoustic reflex was nonreactive in all 10 ears tested. Positive findings in CT scan were 88.89%. CT pneumoencephalography or MRI provided useful information of diagnosis in those with negative findings of place CT scan. CONCLUSION: ABR should be used as a routine test for patients with sudden deafness. When ABR was abnormal, CT scan around the internal auditory meatus was needed. PMID- 12761905 TI - [Facial paralysis in temporal bone trauma]. AB - OBJECTIVE: To study the cause and treatment of facial paralysis in temporal bone trauma. METHODS: A retrospective review of 28 patients with facial paralysis resulting from temporal bone trauma was conducted, House-Brackmann(H-B) system was used to evaluate the recovery of facial nerve function, U test was employed for statistic analysis. RESULTS: Twenty-six cases underwent following up facial nerve function of all patients was recovery partly after surgery, 46% cases reached H-B II, 84% cases reached H-B III, comparing facial nerve decompression within 4 months of injury to beyond 4 months of injury, recovery to H-B II or more of facial nerve function was 60% and 0%, respectively. The difference was significant(P < 0.05). CONCLUSION: Surgery decompression is effective on facial nerve palsy caused by temporal bone trauma. It is recommended that surgery decompression be carried out as soon as possible since a better prognosis is obtained if it is performed within four months after injury. PMID- 12761906 TI - [Ultrastructure changes of the olfactory epithelium of the patients suffering from dysosmia caused by the chronic sinusitis]. AB - OBJECTIVE: To observe the ultrastructural changes of olfactory epithelium (OE) in patients suffering from dysosmia caused by chronic sinusitis. METHODS: The specimens of olfactory epithelium were obtained from 35 patients operated for chronic sinusitis accompanied by dysosmia. According to the results of light microscope (LM) examination, the OE was divided into three groups by the types of pathological changes: normal, atrophic and respiratory epithelium metaplasia(REM). Transmission electron microscope was used to observe the ultrastructural changes of each group. RESULTS: Under the LM, the surface ultrastructure of the OE showed some abnormal changes: (1) surface microvillus of the supporting cells disappeared; (2) olfactory vesicle changed their shape due to vacuolization; (3) disappearance of canaliculus structure in the olfactory vesicle; (4) the olfactory cilia changed the shape or reduced; some of the reduced cilia underwent metaplasia. The ultrastructural changes of atrophic OE included: (1) minor and moderate atrophy: the organelles and the membrane-limited electron dense vesicles on the upper section of the supporting cells obviously decreased or disappeared, even underwent vacuolization. The basic cell degenerated; (2) serious atrophy: the turbidity of the cell structure, even double cell structure, the nuclei of the cell aggregated as the plaque and vesiculose change or karyopyknosis. As for the cytoplasm, there were the dilation of the ERs, turgidity of the mitochondrion, the disarrangement, diminution and vacuolization. Fasciculate cilia were distributed separately in the REM group. CONCLUSION: There is a positive relationship between the atrophy degree and the degree of the abnormal ultrastructural changes of the OE. The ultrastructural changes of OE in patients suffering from dysosmia caused by chronic sinusitis may provide reference for assessment of the treatment of dysosmia. PMID- 12761907 TI - [Clinicopathologic analysis of rhinoscleroma]. AB - OBJECTIVE: To investigate the clinicopathologic characteristics and reason of tissue injury caused by klebsiella rhinoscleromatis(KR) infection. METHODS: Twenty-four cases of rhinoscleromas in the nasal and pharyngeal region in this hospital from March 1983 to March 1998 were studied retrospectively. Warthin Starry (W-S) stain was used in all cases and CD43, CD20, CD68, Lysozyme mark were observed in eleven specimens by ABC immunochemical method. Two cases were studied further with transmission electron microscopy. RESULTS: A great number of KR were found in Mikulicz cells and were confirmed further in the phagosomes in cytoplasm of Mikulicz cells, where a small number of endoplasmic reticulums and lysozymes was squeezed to the side of cells. A few of T-lymphocytes was found within the granuloma. CONCLUSION: Cellular immunity function of the patient with rhinoscleroma was depressed and the response to immunization of host which regards to KR was the major cause of tissue injury. PMID- 12761908 TI - [Correlation of tumor microvessel density with prognosis in laryngopharyngeal malignant melanoma]. AB - OBJECTIVE: To investigate the relationship between microvessel density and clinicopathology, as well as the prognosis in pharyngo-laryngeal malignant melanoma. METHODS: Specific endothelial cell markers with immunohistochemistry and microvessel density with morphometry in 28 cases of laryngopharyngeal malignant melanoma. RESULTS: There was no correlation between the microvessel density (MVD) and the tumor size, Clark grade, as well as Breslow grade; however, significant correlation was found between the MVD and the AJC's grade, as well as the proliferating cell nuclear antigen (PCNA) labelling index. Microvessel counts were associated with overall survival by Kaplan-Meier analysis. An average vessel count of less than 36.5 (x 200) suggested a better survival, but a higher vessel count of more than 36.5 (x 200) showed a trend to worse the overall survival. CONCLUSION: The results suggest a significant relationship between MVD and prognosis; moreover, MVD may be a useful prognostic indicator in laryngopharyngeal malignant melanoma. PMID- 12761909 TI - [Rapid construction of directional cDNA library from human nasopharynx]. AB - OBJECTIVE: To construct a directional cDNA library from human adult nasopharynx by SMART (switching mechanism at 5' end of RNA transcript) technique. METHODS: The total RNA was separated from human adult nasopharynx epithelial tissue and the first-strand cDNA was synthesized through reverse transcription by a modified oligo(dT) primer(contained sfi IB site) while the SMART oligonucleotide(contained sfi IA site) was utilized as a template so that the first-strand cDNA could be extended over the 5' end of mRNA. The double-strand cDNA was amplified by LD PCR(long-distance PCR) with the above two primers and then digested by sfi I (IA & IB) restriction enzyme. After cDNA size fractionation through CHROMA SPIN column, the double-strand cDNA was ligated into the sfi I-digested lambda TripIEx2 vector and then the recombinant DNA was packaged in vitro. RESULTS: The unamplified human adult nasopharynx cDNA library consists of 1.5 x 10(6) independent clones in which the percentage of recombinant clones is about 100%. The titer of the amplified cDNA library is 3.8 x 10(9) pfu/ml and the average exogenous inserts of the recombinants is 1.5 kb. CONCLUSION: These results shows that the human adult nasopharynx cDNA library has an excellent quality and lays solid foundation for screening and cloning new tumor suppressor genes of nasopharyngeal carcinoma(NPC) and tissue-specific genes of human nasopharynx. PMID- 12761911 TI - [Anatomic characteristics of lingual artery and midline glossectomy]. AB - OBJECTIVE: To investigate the relationship between anatomic characteristics of lingual artery and midline glossectomy, for the purpose of increasing operative safety. METHODS: (1) 16 extraoral dissections of the submandibular region were performed on 8 human cadavers and to observe the origin and distribution of lingual artery. (3) The lingual artery was divided into three segments, and the length and adjacent organs were observed and measured. RESULTS: The whole length of lingual artery was (92.88 +/- 13.53) mm; At points of foramen cecum of the tongue, 5 mm and 10 mm prior to foramen cecum, 10 mm behind foramen cecum, and at adjacent part between root of the tongue and epiglotic vallecula, the distances between lingual artery and midline of the tongue were separately measured. The distances were (11.00 +/- 4.95) mm, (8.56 +/- 3.51) mm, (7.78 +/- 3.15) mm, (13.00 +/- 3.28) mm, (15.22 +/- 3.11) mm. The distances between lingual artery and the surface of tongue were (21.4 +/- 5.59) mm, (22.11 +/- 6.74) mm, (22.56 +/ 7.81) mm, (16.56 +/- 5.57) mm, (2.56 +/- 0.73) mm respectively. CONCLUSION: During the midline glossectomy, the lingual artery is easier to be damaged at the position where the root of tongue adjoining epiglottic vallecala. The operation should be done within 10 mm below the surface, 7 mm lateral to the midline, to avoid the damage of lingual artery. PMID- 12761910 TI - [Human papilloma virus infection and expression of p16 protein in laryngeal papilloma and laryngeal carcinoma]. AB - OBJECTIVE: To evaluate the role of human papilloma virus (HPV) infection and inactivation of p16 gene in laryngeal papilloma (LP) and laryngeal squamous cell carcinoma (LC). METHODS: HPV consensus primers direct in situ polymerase chain reaction (ISPCR) and immunohistochemical method were applied to detect the presence of HPV genomes (1, 6, 8, 11, 13, 16, 18, 30, 31, 32, 33, 45, 51) and the expression of p16 protein respectively in 93 cases of formalin-fixed, paraffin imbedded specimens, which contained 46 cases of LPs [adult-onset laryngeal papilloma (ALP) 21, juvenile-onset laryngeal papilloma (JLP)25], 26 cases of LCs, 6 cases of normal tissues adjacent to carcinoma, and 15 cases of vocal noduli. RESULTS: (1) The difference of positive rates of HPV-DNA in JLP group (84%, 21/25) and other groups were statistically significant (chi 2 test, P < 0.05). The difference of positive rates of HPV-DNA in ALPs(38.1%, 8/21), in LCs(19.2%, 5/26), in vocal noduli(0%, 0/15), and in normal tissues adjacent to carcinoma(0%, 0/6) were not significant statistically (chi 2 test or Fisher's exact probability test, P > 0.05). (2) The positive rates of expression of p16 protein in ALP group(57.1%, 12/21) and LC group(38.5%, 10/26) were significantly lower than that in vocal nodule group(93.3%, 14/15), in JLP group(88%, 22/25), and in normal tissues adjacent to carcinoma group (100%, 6/6) (chi 2 test or Fisher's exact probability test, P > 0.05). There were no significant differences of positive rates of expression of p16 protein between ALP group and LC group, and between JLP group and vocal nodule group (chi 2 test, P > 0.05). (3) In LPs, the difference of positive rates of p16 protein expression between HPV positive cases and HPV negative cases was significant statistically (chi 2 test, P < 0.05). In LCs, there was no difference in p16 protein expression rate between the two teams(Fisher exact probability test, P > 0.05). CONCLUSION: The pathogenesis of JLP is closely associated with HPV infection and not associated with the inactivation of p16 gene. Conversely, the pathogenesis of ALP and LC is associated with the inactivation of p16 gene and not associated with the HPV infection. PMID- 12761912 TI - [Pharyngeal passage tube treatment for obstructive apnea syndrome]. AB - OBJECTIVE: To investigate the therapeutical effect of pharyngeal passage tube for OSAS. METHODS: Fifty-seven patients were treated during May 1995 to August 1999. All patients were examined by GKD-405 A polysomnography for apnea index (AI), hypnea index (HI), AHI (AI + HI) at cetera 7 items index before and after pharyngeal passage tube treatment. RESULTS: After treatment, the longest time of apnea shortened from (54.82 +/- 20.83) s to (25.74 +/- 9.50) s, the AHI lessened from 70.82 +/- 18.06 to 30.00 +/- 10.10, the oxygen desaturation increased from (62.36 +/- 11.53)% to (78.68 +/- 12.09)%. After treatment, the recorded parameters showed obvious therapeutical effect (P < 0.05-0.001, t values is from 2.20 to 15.29, the snore loudness of all patients dropped from (84.32 +/- 18.51) dB to (32.64 +/- 10.16) dB. The therapeutical successful rate was 87.72%. The long-term use rate (over 6 months) was 72.73%. CONCLUSION: The pharyngeal passage tube has obvious effect for patients suffering from severe OSAS. It is recommended for the treatment of OSAS as a conservative method. PMID- 12761913 TI - [Study on mutations in the connexin 26 gene among Chinese with nonsyndromic hearing loss]. AB - OBJECTIVE: To study the relation between nonsyndromic hearing loss in Chinese and mutations in connexin 26 (Cx 26) gene and to explore the pathogenic mechanism. METHODS: One hundred and thirty-eight individuals from thirty-five pedigrees with nonsyndromic hearing loss, 99 children with sporadic nonsyndromic hearing loss and 100 normal adults as control were collected in present studies. The Cx 26 coding sequence was screened by single strand conformational polymorphism (SSCP) and analyzed by direct sequencing when SSCP shifts were observed. RESULTS: Five SSCP shifts in 2 pedigrees were observed. Homozygous deletion C at position 233 235 of Cx 26 cDNA, which resulted in frameshift mutation, was found in 2 pedigrees with nonsyndromic hearing loss. CONCLUSION: The hot-spot mutations of Cx 26 gene in Chinese with nonsyndromic hearing loss may be different from other ethnic groups. The 233-235 delC homozygous mutation of Cx 26 cDNA can result in autosomal recessive nonsyndromic hearing loss in Chinese population. PMID- 12761914 TI - [Auricular cartilage palisade technique for repairing tympanic membrane perforation]. AB - OBJECTIVE: To study the auricular cartilage palisade technique for repairing tympanic membrane perforation. METHOD: Sixty-six cases of large tympanic membrane perforation were treated with auricular cartilage. The results were compared with that of temporal fascia repairing in 60 ears. RESULTS: The closure rates of tympanic membrane perforation were 92.4% in cartilage group and 80% in temporal fascia group, respectively. There was no significant difference between two groups in hearing improvement. The cartilage is quite suitable for repairing large perforation of the tympanic membrane and for treating adhesive otitis media. CONCLUSION: The auricular cartilage palisade technique is an ideal method for repairing tympanic membrane perforation. PMID- 12761915 TI - [Endoscopic surgery for nasal inverted papilloma]. AB - OBJECTIVE: To study and perfect the endoscopic surgery for nasal inverted papilloma(IP) and to decrease the recurrence after operation. METHODS: Forty seven cases who were diagnosed as nasal IP were divided into two groups according to the tumor extension and operated upon and observed for 12-40 months (average 24.6 months). 51% of all cases had been operated on for 1-3 times before the treatment. Thirty-six cases in which the tumor had invaded the lateral nasal wall, middle meatus, partial or total ethmoid and/or sphenoid and infiltrated around the natural ostium of the maxillary sinus had performed transnasal endoscopic surgical procedures(TNESP). Another group (9 cases) in which involvement of the IP had further extended, into the maxillary sinus, had been treated by combination transnasal and Caldwell-Luc endoscopic surgical procedures (CTESP). In order to decrease bleeding during the operation, the body and bases of the IP were irradiated with YAG laser. The tumor was removed using the shaver system. The bony attachment of the IP and neighboring tissues were irradiated in all cases. RESULTS: Forty-five (95.7%) cases were not recurrent. The recurrence of the IP was identified in two cases (4.3%). But in 6 cases (12.8%), during postoperative follow-up management (1-3 months postoperatively) small mass of the IP (pathologically identified) were observed and removed and irradiated with YAG laser immediately. The recurrence was not observed in the one year follow-up period. CONCLUSION: Endoscopic surgical technique combining the irradiation of YAG laser and the application of shaver system is recommended. This procedure was effective and led to a decreased recurrence rate. PMID- 12761916 TI - [Subtotal laryngectomy reconstructed by platysma myocutaneous flaps]. AB - OBJECTIVE: To study the feasibility, surgical techniques and the results of laryngeal function preservation during subtotal laryngectomy in laryngeal carcinoma. METHODS: A retrospective review was made of 13 patients treated by subtotal laryngectomy and laryngeal reconstruction from 1991 to 1996. Eleven patients were male and 2 were female. The age ranged from 46 to 72 years, with a mean of 53.25. All cases were of stage IV. The essence of this operation is to reconstruct the new larynx with two platysma myocutaneous flaps following subtotal laryngectomy. RESULTS: The success rate of voice rehabilitation was 100%. Among them, the acceptability rating scores in 11 cases were 7 and the intelligibility scores ranged from 95% to 98%. The other two cases had acceptability 5 and intelligibility form 90% to 94%. The three-year survival rate was 84.6% (11/13), 5-year survival rate was 76.9% (10/13). CONCLUSION: This form of laryngeal reconstruction in subtotal laryngectomy is helpful to restore laryngeal function and decrease the rate of total laryngectomy. The procedure is easy, safe and applicable to patients treated by subtotal laryngectomy. PMID- 12761917 TI - [Postlaryngectomy peristomal recurrence]. AB - OBJECTIVE: To study the etiopathology, diagnosis and management of peristomal recurrence after laryngectomy, and the prevention of peristomal recurrence. METHODS: Retrospective analysis of clinical management and outcomes of 16 cases of peristomal recurrence. RESULTS: The subglottic extension of laryngeal cancer and peritracheal metastasis may result in peristomal recurrence. The stomal resection with mediastinal dissections and the use of the pectoralis major myocutaneous flap were done. The survival period of 11 cases surgically treated was longer than that of 5 cases irradiated or untreated. CONCLUSION: Surgical management would improve the survival, but overall prognosis is dismal and attention should be focused on prevention. It is evident that early diagnosis of stomal recurrences will yield good results. PMID- 12761918 TI - [Study on voltage-sensitive current of spiral ganglion cells in mice organ of Corti culture]. AB - OBJECTIVE: To investigate the property of voltage-sensitive current in cochlear spiral ganglion cells of the C57BL/10J mice, an inbred strain which develops early onset hearing loss. METHODS: Organotypic cultures of organ of Corti were prepared from neonatal mice 0-5 days of age. Whole-cell current and voltage clamp techniques were used to study Na+, K+ and Ca2+ currents of the spiral ganglion cells in culture. RESULTS: Cultures were maintained for 8-48 hours before use. Ganglion cells were identified first through their anatomical positions and finally through fast negative Na+ current. Spontaneous action potentials were recorded from some ganglion cells (4 out of 39). When present, spontaneous rates were around 20 spikes/sec, and might be as high as 135 spikes/sec. The mean resting potential was (-55 +/- 5) mV (n = 39). Under voltage clamp conditions, transient inward currents (negative) and outward (positive), steady-state voltage dependent currents were recorded in normal HBSS. Rapid inward currents were totally blocked by 300 nM TTX applied locally to the culture. Inward currents recovered quickly after TTX wash out suggesting that the transient inward current was carried by Na+. The mean maximum amplitude of Na+ current was (-2.0 +/- 1.1) nA (n = 39) recorded in HBSS. Adding TEA (10 mmol/L) and 4-AP (0.15 mmol/L) to the bath solution or replacing K+ with Ca+ in the pipette solution partly blocked the sustained outward current. This suggests that the outward current was carried by K+. The mean maximum amplitude of K+ was (3.0 +/- 1.3) nA (n = 39) with 140 mM K+ in the pipette. Inward Ca2+ current was recorded in Ba2+ solution which mean peak amplitude was (-1.0 +/- 0.7) nA (n = 20). Ca2+ currents were reversibly blocked by 100 microM Cd2+. CONCLUSION: Whole cell recordings from spiral ganglion neurons can be obtained from organotypic cultures of the organ of Corti. Fast Na+ current, sustained K+ current and L-type Ca2+ current were recorded in the spiral ganglion cells cultured for 1-2 days. Whole cell recording showed that cochlea spiral ganglion cells can generate spontaneous action potential one day after birth and the firing rates could reach levels equal to those recorded in vivo. PMID- 12761919 TI - [The relationship between cell proliferation and apoptosis of vestibular epithelium in the developing guinea pig]. AB - OBJECTIVE: The cell proliferation and apoptosis of vestibular epithelium in normal developing guinea pig were observed to elucidate the relationship and effect between proliferation and apoptosis in developing vestibular epithelium. METHODS: Twenty Guinea pigs with different developing ages were used in our studies. Cell proliferation was detected by proliferation cell nuclear antigen (PCNA) immunocytochemistry while apoptosis was detected by TdT-mediated deoxyuridine triphosphate-biotin nick end labelling (TUNEL) method and transmission electro-microscopy. RESULTS: PCNA staining had been detected in supporting cells but not in hair cells, and the positive cells were gradually decreased with ageing. The differentiated hair cells and supporting cells underwent apoptosis in vestibular sensory epithelium, and TUNEL-positive cells were also markedly decreased with ageing. After birth, TUNEL-positive hair cells were located at the top of ampulla. CONCLUSION: During the development of guinea pig, cell proliferation as well as apoptosis events diminish contemporaneously. The balance between proliferation and apoptosis may regulate the normal number of cells, and is related to guarantee the normal structure and function of inner ear. PMID- 12761920 TI - [Beta-actin changes during hair cell regeneration in the chick basilar papilla]. AB - OBJECTIVE: To investigate the changes of beta-actin and beta-actin mRNA expression during hair cell regeneration in the chick basilar papilla(BP) following gentamicin ototoxicity. METHODS: Thirty-six chicks were given muscular injection of gentamicin at dosage of 100 mg/kg.d for 10 consecutive days, and twelve additional chicks received injection of normal saline instead of gentamicin as controls. At 1, 3, 7, 14, 21 and 28 days after the treatment, six chicks in experimental group and two chicks in control group were sacrificed. Basilar papillas were prepared for immunohistochemistry and in situ hybridization study. beta-actin and beta-actin mRNA expression in the damaged region were histologically analyzed by computer image system. RESULTS: beta-actin immunoreactivity of BP changed significantly after treatment with gentamicin. From the first day to the fourteenth day after gentamincin, beta-actin immunoreactivity turned positive gradually. It approximated the normal response level at the fourteenth day after treatment of gentamincin. Then increasing rate of beta-actin immunoreactivity positive response level slowed from the fourteenth day after the treatment. At seventh day after treatment, beta-actin mRNA expression in BP increased to peak value, which preceded the peak expression of beta-actin in the regenerated hair cells. CONCLUSION: Nascent hair cells seem to mature 14 days after hair cell damage, which may play a role in auditory functional recovery. PMID- 12761922 TI - [Response property of tone burst transmission of partially implantable middle ear implant in vivo]. AB - OBJECTIVE: To investigate the characteristics of tone burst transmission of the middle ear implant(MEI). METHODS: Ten adult rabbits with normal hearing were used. A silver ball electrode was placed at the round window membrane to measure cochlear microphonics (CM) when sounds were presented before and after the implantation of the MEI. The tone burst was the acoustic stimulus. RESULTS: Frequency response functions without and with the MEI in rabbit model were calculated, and their amplitude-frequency curves ranging from 0.5 to 6 kHz resembled each other in shape with gain factors about 15 dB at the frequencies above 1 kHz. CONCLUSION: The results of this study indicated that the frequency response of the device fitted the speech transmission and it had high fidelity with little noise. PMID- 12761921 TI - [Detection of GABAA alpha 2 mRNA in rat vestibular end organ with in-situ hybridization]. AB - OBJECTIVE: To investigate the effect of the gamma-aminobutyric acid (GABA) on rat vestibular end organ. METHODS: In the rat inner ear paraffin slides, GABAA receptor alpha 2 subunit mRNA in vestibular end organ has been detected with in situ hybridization. Digoxigenin-GABAA alpha 2 cDNA prob (549 base pair), Anti Digoxigenin-AP (Fab fragments) and BM Purple AP Substrate (precipitating) have been used. RESULTS: GABAA receptor alpha 2 subunit mRNA has been found in all vestibular ganglion cells and nerve ending surrounding type I hair cells in rat's cristae ampullaris. GABAA alpha 2 has not been found in hair cells and supporting cells. As a positive control, GABAA alpha 2 mRNA has been found in Purkinje cells and granule cells in rat's cerebellum. GABAA alpha 2 has not been found in OMP negative control, non-prob negative control and non-anti-digoxingenin control in inner ear and cerebellum slides. CONCLUSIONS: GABAA alpha 2 receptor has been found in vestibular afferent nerve endings and vestibular ganglion cells. It strongly indicates that GABA is an afferent nerve transmitter in the vestibular end organ and plays an important role in the vestibular afferent nerve transmission. PMID- 12761923 TI - [Surgery of the skull base assisted by sinus endoscope]. AB - OBJECTIVE: To explore the feasibility and clinical value of the endoscopic operations around the skull base. METHODS: A retrospective study was made of 44 cases treated by endoscopic operations. Twenty-five cases underwent intranasal endoscopic operations. RESULTS: Forty of 44 cases were cured by one-stage surgery. One case of cartridge foreign body clamped in clivus was not successfully removed; only window operation and puncture biopsy were done in another case of pituitary adenoma; high cranial pressure has not been completely reduced after operation in the third case with relapsing craniopharyngioma a companied with obstructive hydrocephalus. One case of sphenoidal malignant adenoma involving saddle based and extended laterally was partly resected. Complications were follows: cerebrospinal fluid rhinorrhea in 2 cases, diabetes insipidus in 1 case, all of them were cured by conservative treatment; nasal septum perforation in 1 case, without any treatment. CONCLUSIONS: The endoscopic skull base surgery by intranasal approach is feasibility. Intranasal approach has direct path, slight wound and no wound in face. Combined with operation microscope, sinus endoscope can make up for its limits. PMID- 12761924 TI - [Significance and expression of transforming growth factor beta in human nasal polyp tissue]. AB - OBJECTIVE: To explore the pathogenesis of nasal polyposis and the expression of transforming growth factor beta (TGF-beta) in human inflammatory nasal polyps. METHODS: TGF-beta 1-3 in nasal polyp tissues and inferior turbinate mucosa of twenty-five polyposis patients were detected with immunohistochemistry alkaline phosphatase and anti-alkaline phosphatase (APAAP) method. The inferior turbinate mucosa of eight healthy volunteers were selected as control. Six polyp tissues were estimated with double immunolabeling and Western-blot analysis to compare the characterization of the TGF-beta isoforms expression and the proportion of macrophages and eosinophils in nasal polyp tissues. RESULTS: The expression of TGF-beta 1-3 in nasal polyps was significantly higher than that in nasal mucosa and indetecable in nasal mucosa from healthy volunteers; TGF-beta 1 was the main isoform detected in nasal polyps; TGF-beta positively was accompanied by numerous macrophage and eosinophil infiltration. CONCLUSIONS: TGF-beta mainly TGF-beta 1 is strongly expressed in nasal polyps and its mucosa, where it could be produced by macrophages and eosinophils. TGF-beta could induce modification of epithelium and connective tissue and therefore be involved in the pathogenesis of nasal polyposis. PMID- 12761925 TI - [Empty nose syndrome]. AB - OBJECTIVE: The aim of this article is to present the concept of empty nose syndrome(ENS) and help ENT doctors take care of regular nasal turbinate surgery. METHODS: Fourteen patients who was diagnosed as ENS in our department were reviewed retrospectively. All patients had undergone various forms of nasal surgery (all had had turbinectomy). Their age ranged from 13 to 52 years. All patients were treated conservatively, among these patients 5 subjects who had more serious symptoms received submucous and subperiostal nasal implantation with ilium. RESULTS: All patients had nasal obstruction and dryness of nasal cavity, nasopharynx and oto-pharynx in 6 months--5 years after their first nasal surgery, some presented symptoms of depression. Nasoscope examination showed all patients had a cylindrically enlarged nasal cavity. Conservative treatment was effective in most cases. The effectiveness of operative treatment was encouraging during short-term follow-up. CONCLUSION: Extensive turbinectomy may cause secondary nasal mucosal atrophy and a series of subsequent symptoms. The presentation of this concept is to remind the ENT doctors prudently performing turbinectomy to avoid the occurrence of irreversible injury to the nasal cavity. PMID- 12761926 TI - [Misdiagnosis in isolated sphenoid sinus diseases]. AB - OBJECTIVE: To improve the correct diagnosis for isolated sphenoid sinus diseases. METHODS: Twelve patients with isolated sphenoid disease from 1991 to 1998 were reviewed. The factors of misdiagnosis were discussed. RESULTS: In this group of 12 patients with isolated sphenoid diseases, 8 failed to be diagnosed at the first time of medical consultation. The misdiagnosis included optic neuritis, neuralgia, ophthalmitis, pulmonary tuberculosis, epistaxis and epilepsy. CONCLUSIONS: The main causes of misdiagnosis were: low incidence of the disease, nonspecific clinical symptoms, and unawareness of the doctor. CT or magnetic resonance imaging (MRI) can provide much more informations about the diagnosis. PMID- 12761927 TI - [Titanium miniplate fixation in head and neck cancer surgery]. AB - OBJECTIVE: To evaluate the rigid fixation with miniplates in head and neck cancer surgery. METHOD: A consecutive series of 40 patients with head and neck tumor underwent rigid fixation with the Martin titanium miniplates system in a 6 years period. The miniplates were used to fix bone graft reconstruction following mandible resection (n = 14), mandibulotomies for oral and oropharyngeal surgical access (n = 12), maxilla swinging for nasopharyngeal access (n = 11) and other purposes (n = 3). Twenty-three of all cases(57.5%) received perioperative radiotherapy with average doses of 60Gy at the osteotomy site. RESULTS: Eleven of the 40 patient (22%) developed plate related complication which included 5 infections, 3 mandibular osteonecroses, 1 plate exposure, 1 screw loose and 1 nonunion. Complications were more likely to occur in patients with perioperative radiation than patients without (39.1% versus 11.8%, P = 0.079). Except one case, All the uncomplicated dentulous patients demonstrated stable and good occlusion. Oral feeding usually started on the 3-4th day after surgery. There was no need for intermaxillary fixation. CONCLUSION: The miniplate provided stable mandibular or maxillary fixation and accurate repositioning, eliminated the need for intermaxillary fixation. The malleability and versatility of miniplate make it unique in bone graft shaping for mandibular reconstruction. Complication seems to be related to perioperative radiotherapy. Median or paramedian mandibular osteotomy, which is non-radiated region in most radiotherapy cases, is advocated. PMID- 12761928 TI - [The reconstruction of laryngeal function in subtotal laryngectomy by pedicled flaps]. AB - OBJECTIVE: To explore the possibility of subtotal laryngectomy in the treatment of advanced laryngeal cancer and selection of reconstruction. METHODS: Forty patients were treated surgically by subtotal laryngectomy with preservation of arytenoid cartilage and perichondrium. The pedicled flaps between cricoid cartilage and arytenoid cartilage were sewed up. The new larynx was reconstructed by suturing the cricoid or trachea to the hyoid bone or the tongue base. RESULTS: The 3 and 5 year survival rates were 85.0% and 76.2% respectively. Decannulation rate was 92.5%. CONCLUSION: The reconstruction of laryngeal function in subtotal laryngectomy by pedicled flaps not only is safe and beneficial to the patients with the cancers above the cricoid, but also improves the quality of patient's life. PMID- 12761929 TI - [A clinical evaluation on reconstruction in extended partial laryngectomy for transglottic cancer]. AB - OBJECTIVE: Transglottic cancer has been a definite indication for total laryngectomy in decades, compromising function of the larynx. The purpose of this study is to present a new radical surgery of extended partial laryngectomy and reconstruction to restore the essential function of larynx. METHODS: Forty-one cases of transglottic cancer were treated by extended vertical laryngectomy (26/41), extended frontolateral laryngectomy (5/41) and subtotal laryngectomy (10/41); the defects of larynx were reconstructed by rotary door myocutaneous flap in twenty-seven cases, by sternohyoideus flap in seven cases, by osteomuscular flap in five cases and by sternocleidomastoid flap in two cases. RESULTS: The 3 and 5 year survival rates were 85.7% (30/35) and 74.1% (20/27) respectively, those for stage III were 84.6% (22/26) and 76.2% (16/21), for stage IV were 3/4 and 1/2 respectively. All cases resumed acceptable voice, 92.7% (38/41) and enjoyed satisfactory phonation. The over-all decannulation rate was 87.8% (36/41), those with rotary door myocutaneous flap was 96.3% (26/27), with sternohyoideus flap was 5/7, with osteomuscular flap was 4/5 and with sternocleidomastoid flap was 1/2. All patients had normal swallow function. CONCLUSION: Extended partial laryngectomy is a kind of radical operation in selected cases of transglottic cancer. The method of reconstruction with rotary door myocutaneous flap can improve curative effect and the essential functions of the larynx. PMID- 12761930 TI - [Multiple factors analysis of intraoperative bleeding and recurrence of juvenile nasopharyngeal angiofibromas]. AB - OBJECTIVE: To study the effect of surgical treatment of juvenile nasopharyngeal angiofibromas and the factors influencing intra-operative bleeding and recurrence. METHODS: Thirty-four patients with juvenile nasopharyngeal angiofibromas treated surgically in Chinese PLA General Hospital between 1986 and 1999 were studied retrospectively. The relationship between surgical treatment, intraoperative bleeding, recurrence and age, duration, staging, preoperative treatment, times of previous operation, operative approaches was statistically analyzed. RESULTS: The tumors were totally resected in 30 patients, and five patients(16.7%) recurred during a mean follow-up of 77 months. The mean recurrence time after operation was 3.2 months (1-6 months). The amount of intraoperative bleeding correlated well with the age, duration, staging, preoperative treatment, and surgical approaches (P < 0.05), but not with times of previous operation (P > 0.05). Recurrence was not correlated with the age, duration, preoperative treatment and surgical approaches (P > 0.05), but correlated with staging (P < 0.05). CONCLUSION: The factors significantly influencing the intraoperative bleeding were the age, duration and staging. Recurrence was correlated with the tumor stages, incomplete resection or the exceedingly malignant activity of the tumor. Radiotherapy, with a dose of 30Gy, is an adjuvant therapy for incompletely resected, or residual tumors. PMID- 12761931 TI - [The diagnosis and treatment of large cervical vascular injury]. AB - OBJECTIVE: To study the diagnosis and treatment of large cervical vascular injury, and to improve the treatment. METHODS: Clinical data of 18 cases of large cervical vascular injury between 1974 and 1999 in the Medical College Second Affiliated Hospital of Zhejiang University were analyzed retrospectively. RESULTS: The following injuries were found, including common carotid artery involvement in 7, external carotid artery in 4, internal carotid artery in 2, subclavian artery in 1, internal jugular vein in 4. The type of injury included incision injury in 8, laceration injury in 5, break injury in 2, inflammatory injury in 2, tumor injury in 1. Fifteen cases were diagnosed by active bleeding of wound in neck, 2 cases by color Duplex ultrasound, 1 case by exploration. Operation included vascular repair in 8, vascular ligation in 10. The result showed that 14 patients were cured, one patient developed light hemiplegia, one patient performed amputation of the arm due to necrosis after subclavian artery ligation, two patients died, the mortality rate was 11.1%. CONCLUSION: Active profuse bleeding from the neck wound is the basis of diagnosis basis. Rational surgical treatments are needed immediately. PMID- 12761932 TI - [Long-term follow-up of patients with obstructive sleep apnea syndrome treated with uvulopalatopharyngoplasty]. AB - OBJECTIVE: To study the long-term outcome of uvulopalatopharyngoplasty (UPPP) in the treatment of obstructive sleep apnea syndrome (OSAS). METHODS: Long-term follow-up for 32 cases of patients with OSAS was conducted in 3 years after UPPP. The respiratory disturbance index (RDI), body mass index (BMI), blood pressure (BP), fasting blood glucose (FBG), triglyceride, cholesterol were followed. RESULTS: RDI was reduced markedly from 58.1 +/- 25.5 to 38.7 +/- 15.3 at 3 years after UPPP. The total effective rate was 87.5% after one month. 55.2% after 3 years. Their BMI, BP, FBG, triglyceride and cholesterol were also reduce significantly. CONCLUSION: UPPP can improve the hypoxemia of patients with OSAS, reduced the BP, FBG, BMI, improve the lipid metabolism. Long-term outcomes of UPPP were possibly related to the preoperative BMI, RDI, the skeletal structure of the upper airway and postoperative BMI change. After UPPP, long-term follow-up is recommended because some initially successfully treated patients with OSAS will relapse in the later follow-up. PMID- 12761933 TI - [Management of irregular shaped airway foreign body in children]. AB - OBJECTIVE: To explore proper operative methods for removal of irregular shaped foreign bodies in children. METHODS: Forty-eight emergency cases of irregular shaped airway foreign bodies (plastic ball-pen cap, mini-toys, whistle, pin, thumbtack, milk tooth, pen, steel ball, stone of apricot, chicken bone and screw nail) from Jan. 1998 to Nov. 2000 have been reviewed, especially the operative procedure for removing foreign bodies. The youngest patient was 9-month old and eldest 14 year-old. The average age was 8.3 year-old. The male female ratio was 2.4 storz tracheoscope (Made in German) was applied. RESULTS: Forty-six of the 48 foreign bodies were removed through tracheoscope. The foreign bodies slipped off at the level of the glottis in 3 cases, they were then removed from the opposite bronchus (respiratory difficult syndrome developed in these cases); a thumbtack was removed by tracheotomy; one stone of apricot was removed through. CONCLUSIONS: An accurate pre-operative diagnosis is most important in operative management of cases of irregular shaped foreign bodies. General anesthesia, is usually. If the foreign body is too big to pass through the glottis, removing by tracheotomy should be considered. PMID- 12761934 TI - [Ultrastructure of surfactant-like multilamellar bodies in the human nose]. AB - OBJECTIVE: To study the distribution of the surfactant-like multilamellar bodies in the human nose. METHODS: Fourteen specimens obtained from the inferior turbinate, middle turbinate, uncinate process and maxillary sinus were examined under the electron microscope. RESULTS: The surfactant-like multilamellar bodies were identified in the epithelium on the lateral wall and maxillary sinus. CONCLUSION: The existence of surfactant-like multilamellar bodies in the human lateral nasal wall and maxillary sinus were confirmed. It is necessary to have further study for its effect in the nose both physiologically and pharmacologically. PMID- 12761935 TI - [Observation of nasal mucosal cilia ultrastructure of Kartagener's syndrome]. AB - OBJECTIVE: To report the ultrastructure of nasal mucosal cilia of Kartagener's syndrome and the operative results of 2 cases. METHODS: Both two cases were underwent endoscopic sinus surgery. Transmission and scanning electron microscopic examinations of nasal mucosa for case 1 were performed. RESULTS: All 2 cases presented situs inversus, chronic paranasal sinusitis and bronchiectasis. The electron microscopic examination showed that the shape of nasal cilia and "9 + 2" structure were normal, but the lack of inner dynein arm of cilia and confused arrangement of central pair of microtubules were confirmed. The recurrent of nasal polyps after 1-year follow-up was observed in case 1. CONCLUSIONS: The disturbance of mucociliary clearance due to congenital ciliary structure defect might be the cause of chronic sinusitis and polyps, which might be one of reasons for poor prognosis after operation and we should pay more close attention. PMID- 12761936 TI - [A comparative observation preoperatively and postoperatively of nasal mucosa in chronic sinusitis treated with endoscopic sinus surgery]. AB - OBJECTIVES: To study morphologic and functional mucosal changes both preoperatively and postoperatively in chronic sinusitis (CS) treated with endoscopic sinus surgery (ESS). METHODS: 1. Saccharin test and light microscopic examination of nasal mucosa were taken preoperatively and postoperatively in 32 cases with CS and in 28 normal subjects as control. 2. Scanning and transmission electron microscopy were performed to exam preoperatively and postoperatively the nasal mucosa in 10 cases with CS and in 2 normal cases as control. RESULTS: 1. The preoperative saccharin test time (STT) in patients with CS was significantly longer than that in the controls (P < 0.001), and the postoperative STT became significantly shorter than that in preoperative one(P < 0.001). 2. The pathological changes of nasal mucosa, such as infiltration in inflammatory cells, edema, polypoid formations and pathologic glands were observed preoperatively. The infiltration in inflammatory cells, edema and polypoid formations were significantly released (P < 0.01) at four months after operation, and there was no significant difference comparing with the controls (P > 0.05). However, the pathologic glands were not reduced even after four months postoperatively. 3. The examination of electron microscopy demonstrated that the ultrastructure of nasal mucosa was impaired preoperatively and almost completely recovered at four months postoperatively. CONCLUSIONS: The normal structure and clearance function of nasal mucosa in patients with CS was impaired preoperatively, and the impaired structure and mucociliary clearing function of nasal mucosa were greatly improved after ESS and almost completely recovered at four months postoperatively. The pathologic glands and secretive function of nasal mucosa may need longer period for recovering. PMID- 12761937 TI - [Expression and distribution of Aquaporin 1 in the nasal polyps]. AB - OBJECTIVE: To confirm the expression and distribution of Aquaporin 1 (AQP1) in the nasal polyps and to investigate the relation between AQP1 and the nasal polyp edema. METHODS: fourteen cases of normal inferior turbinates and 26 cases of nasal polyps were used. The expression and distribution of AQP1 in nasal polyps were examined by immunohistochemical technique. RESULTS: The level of AQP1 in the epithelial cells and serous cells from nasal polyps was higher than that in inferior turbinates, and the level of AQP1 in the epithelial cells and cilium cells from inferior turbinate was higher than that in nasal polyps. CONCLUSION: There is close relationship between AQP1 and nasal polyp edema. PMID- 12761938 TI - [Roles of CD43, CD20 and local IgE in the pathogenesis of nasal polyps]. AB - OBJECTIVE: To estimate the roles of CD43, CD20 and total IgE in the pathogenesis of nasal polyps. METHOD: Twenty-six cases of nasal polyps and 20 cases of mucosa of middle turbinates were used in present study. RESULTS: The number of CD43+ cells, CD20+ cells and plasma cells in nasal polyps were significantly more than that in middle turbinates (P < 0.01). The positive rate of local IgE+ cells in nasal polyps was significantly higher than in middle turbinate (P < 0.01). CONCLUSION: There were active cellular immunity and humoral immunity in nasal polyps. Local allergy may play a role in the pathogenesis of nasal polyps. PMID- 12761939 TI - [Intraoperative complications of endoscopic sinus surgery]. AB - OBJECTIVE: To analyze the complications occurring in endoscopic sinus surgery (ESS). METHODS: The complications occurred in 25 cases from 307 patients who accepted ESS, performed by author, were reviewed. RESULTS: The complications were periorbital injury (n = 16), medial rectus injury (n = 2), optic nerve injury (n = 1), retrobulbar haematoma (n = 1), inferior turbinate feared partially (n = 2) and severe hemorrhage(n = 3). CONCLUSION: The causes of complications occurring in ESS were following: lack of experience, hemorrhage, blind operation, unsuitable instrument and reckless in manipulation. In addition, the complications, even serious complications, occurred in the hands of experienced surgeon. PMID- 12761940 TI - [Effects of direct current on vibration of cochlear basilar membrane]. AB - OBJECTIVE: To explore the effects of direct current on vibration of cochlear basilar membrane. METHODS: A small hole with a diameter about 0.4 mm, which had the distance of 2.4 mm from round window niche, was made on cochlear basic turn of guinea pig for measuring the velocity of basilar membrane in vivo. On the superior and inferior edge of the measuring window, holes were made on the scala tympani and scala vestibuli for insert stimulating electrode that was made of platinum-iridium. The basilar membrane vibration was elicited by tone and the effect of direct current stimuli were observed by Laser Doppler interference velocimeter. RESULTS: The velocity of basilar membrane vibration elicited with tone was increased significantly when positive current was delivered to scala vestibuli and the velocity was decreased on the contrary. CONCLUSION: The positive endolymphatic potential is essential for cochlea to transit sound to nerve impulse in physiological condition. It is benefit to enhance the gain of cochlear amplifier when the positive potential is increased on the top of outer hair cells, but the extrinsic negative potential should significantly decrease the gain of cochlear amplifier. PMID- 12761942 TI - [Evaluation of sensorineural hearing loss in childhood]. AB - OBJECTIVE: To explore the clinical and audiological characteristics of sensorineural hearing loss (SHL) with pathological changes both in cochlea and retrocochlear in children and evaluate the relationship between SHL and the lesions in the central nervous system (CNS). METHODS: Three hundred and ten cases (500 ears) of SHL accepted between 1998 and 2000 were studied. The age of patients was ranged from 1 month to 6 years old. According to the evaluation of function of CNS by pediatric neurologist, all cases were divided into two groups: SHL with CNS disease and SHL without CNS disease. Some same age children without hearing loss were subjected as control group. All children were tested using both auditory brainstem responses (ABR) and distortion product otoacoustic emissions (DPOAE). RESULTS: 1. The rate of SHL accompanied with CNS diseases was very high in these children. 2. Patients with kernicterus-cerebral palsy usually had serious hearing loss caused by acoustic nerve lesion at retrocochlear, but their cochlea function was injured slightly. Patients with external hydrocephalus had only slight acoustic nerve lesion at retrocochlear, and patients with other CNS diseases usually had no change for their cochlea function. 3. In the group of SHL caused by cochlea lesion, amplitudes of DPOAE decreased obviously when the threshold of wave V of ABR was up to 60 dB nHL, and amplitudes of DPOAE seriously decreased or disappeared when the threshold of wave V was up to above 70 dB nHL. CONCLUSION: The patients with SHL are usually accompanied with CNS diseases in childhood, and their hearing loss appears very difference from audiological characteristics. We suggest that it is necessary to test both ABR and DPOAE in these patients, and it is important that pediatric neurologist join in audiologist team for our clinical study. PMID- 12761941 TI - [Synergetic protective effects of glial cell line-derived neurotrophic factor combined with neurotrophin-3 in F-actin on hair cell after noise trauma]. AB - OBJECTIVE: To investigate if glial cell line-derived neurotrophic factor (GDNF) combined with neurotrophin-3 (NT-3) provides synergetic protection in filamentous actin (F-actin) on hair cell (HC) from acoustic trauma. METHODS: Guinea pigs were exposed to 4 kHz narrow band noise at 115 dB SPL for 4 h. Test group (n = 12) with a mixture of GDNF (100 ng/ml) and NT-3(2.5 micrograms/ml) or control group (n = 9) with artificial perilymph (AP) was delivered to the scala tympani via a mini-osmotic pump (0.5 microliter/h) for a total of 14 days. Auditory function was assessed by measuring thresholds of auditory brainstem responses (ABRs) elicited by clicks prior to surgery, 3 days after surgery (1 day before noise exposure) and 10 days following noise exposure (before animals were sacrificed), respectively. F-actin, labeled by rhodamine-phalloidin, was examined in the guinea pig cochlea using fluorescence microscopy for quantitative assessment of hair cell damage. RESULTS: There was a statistically significant increase the survival of out hair cell(P < 0.001, P < 0.01) and inner hair cell(P < 0.01, P < 0.01) and decrease in ABR threshold (P < 0.05, P < 0.01) in both the GDNF and NT 3 treated and untreated ear of animals. CONCLUSION: Our findings indicate that GDNF combined with NT-3 may effectively protect the inner ear from noise--induced hearing loss. PMID- 12761943 TI - [The intraoperative application of neural response telemetry with the nucleus CI24M cochlear implant]. AB - OBJECTIVE: To test device integrity and objective auditory reaction for cochlear implant patients intraoperatively. METHODS: Our protocol for intraoperative testing of the implant device includes device electrode impedance test and neural response telemetry (NRT), which measures the electrically evoked auditory nerve compound action potentials (ECAP). RESULTS: We completed electrode integrity tests and NRT intraoperatively on 40 patients with the Nucleus CI24M cochlear implant. We have measured normal implant function on all 40 patients and obtained ECAP results from 39 patients. Out of 33 patients with normal inner ear, typical ECAPs were recorded in 195 electrodes in all 198 testing electrodes(98.5%). In 7 patients with inner ear Mondini dysphasia, affirmative ECAP waveforms were recorded in 26 electrodes in all 42 testing electrodes. The basal electrode ECAP threshold was higher than that of the epical one. CONCLUSIONS: The intraoperative measurements of device function and NRT are quite simple, quick and with high accuracy. Inner ear malformation is an important factor that may affect NRT results. PMID- 12761944 TI - [The role of fibrinolysis in pathogenesis of middle ears adhesions]. AB - OBJECTIVE: To investigate the role of fibrinolysis in pathogenesis of middle ears adhesions. METHODS: The amount of Tissue-type Plasminogen Activator (tPA) of 28 sections from 6 ears with adhesive otitis media (AOM) and of 22 sections from 6 normal ears was examined by Super Sensitive Biotin-Streptavidin (SSBSA) method. Amount of Fibrin of 11 sections from 3 ears with significant adhesions was compared with that of 12 sections from 3 normal ears. Qualitative analysis of light microscopy with computer-assisted image system was employed. RESULTS: In adhesive ears, tPA stains were negative in 11 of 28 sections and faint positive were 10 of 28 sections, while Fibrin stains were positive in 6 of 11 sections and strong positive were in 3 of 11 sections. In normal ears, tPA stains positive were in 8 of 22 sections and strong positive were 10 of 22 sections, meanwhile, fibrin stains were negative in 8 of 12 sections and faint positive were in 3 of 12 sections. Quantitative analysis showed that the amount of tPA was 16.70 +/- 5.11 and 39.84 +/- 6.26 in ears with AOM and normal ones respectively (P < 0.05). CONCLUSION: In adhesive ears the amount of tPA was less than that in the normal ears, whereas, the amount of Fibrin was greater in ears with AOM than that in normal ears. It indicates that fibrinolysis involved in the process of adhesion formation of AOM, which may acts as a key factor. PMID- 12761945 TI - [Surgical treatment of craniofacial recurrent carcinoma]. AB - OBJECTIVE: The patients with craniofacial recurrent cancers after the radiation, chemical and operative therapies were often not able to get effective treatment because their lesions involved in the nervous and vascular structure and the re radiation therapy was not suitable or chemical therapy was not sensitive to the tumor as well. To improve their survival quality, prevent the fatal complication and prolong their life, the surgical treatment for them was advised. METHOD: Six patients were treated surgically. According to the lesion of extension and position, the different incisions were respectively made. The neoplasms of these patients were resected piece by piece because they usually involved in the important structures, such as the internal carotid artery and cavernous sinus. Then, local defect was repaired with different methods on basis of the individual condition. So, this type of operation could be called as non-defined operation. RESULTS: The lesions in all 6 patients invaded the eye, meninges and encephala, skull base, cavernous sinus and other important structures. After surgical intervene, the patients were all more than one year survival except for one of the patients who died of serous bleeding with corrupted rupture of internal carotid artery. Three patients survived for 4, 5 and 12 years, respectively. CONCLUSION: The craniofacial recurrent carcinomas in late stage could be resected by the non-defined operation, which could prolong the patients life and achieve the desired effect. PMID- 12761946 TI - [CT and MRI and clinical study of chordoma in the skull base]. AB - OBJECTIVE: To investigate the diagnostic features of CT and MRI in chordoma of the skull base (CSB) and to estimate their value of clinical application. METHODS: Sixteen patients with CSB were performed CT and MRI examinations of the head, and 13 patients with nasopharyngeal carcinoma (NPC) and 11 patients with pituitary adenoma (PA) which invaded the skull base were selected as control. In addition, 7 cases of chordoma were performed multiple planar reconstruction (MPR) of MRI. RESULTS: MRI was superior to CT in depicting the location and the extent of CSB. The bone destruction at mid-line skull base was demonstrated equally well on both CT and MRI. Based on the location of the tumor displayed on MRI, the CSB could be classified into selloclival, nasopharyngeal, and occipito-temperal types. The T2 weight imaging(T2WI) signal intensity of chordoma was significantly higher than those of NPC and PA (P < 0.01). On MPR images of MRI, oppressed and displaced optic nerves caused by chordoma were noted in 3 cases and encased optic nerves in 4 cases, encased abducent nerves in 7 cases with abducent paralysis and mildly oppressed oculomotor nerves in 3 cases. CONCLUSION: The extremely high T2WI signal intensity is the characteristic of MRI finding in CSB and has great differential value for the tumors of the skull base. The MPR of MRI plays an important role in the surgical treatment for these tumors. PMID- 12761947 TI - [Loss of heterozygosity on chromosome 9p13-23 in microdissected laryngeal squamous cell carcinoma by microsatellite analysis]. AB - OBJECTIVE: Loss of heterozygosity (LOH) studies indicate that allelic loss associated with the development of head and neck of squamous cell carcinoma occurs most frequently at 9p21-22. However, the target of chromosome 9p21-22 loss has been the source of significant debate. A putative tumor suppressor gene, p16, has been identified at the 9p21 location, but genetic alterations of p16 located in this region are unusual. To refine the hot spots of LOH on chromosome 9p13-23 will be helpful to find other putative tumor suppressor genes in laryngeal squamous cell carcinoma. METHODS: The sections of paraffin-embedded tumor tissues were microdissected to enrich for neoplastic cells. LOH on 9p13-23 was analyzed in a set of 42 paired blood and tumor samples using polymerase chain reaction (PCR) with 13 highly polymorphic microsatellite markers. RESULTS: Of the 42 total tumors, 41(97.6%) showed LOH in at least one of the microsatellite markers tested at the chromosome 9p13-23 region. The most frequently deleted marker was D9S162 with in 17 of 19 (89.5%) informative samples. The marker D9S171, which is located in 9p21, had LOH detected in 12 of 15 informative cases (80.0%). LOH at the D9S1748 marker (closest to the p16 gene locus) was detected in 18 of 36 informative cases (50.0%). Fine deletion mapping also revealed two minimal regions of LOH encompassing markers D9S161-D9S171 at 9p21 and IFNA-D9S162 at 9p22 23. CONCLUSION: These findings imply the high frequency of LOH at 9p13-23 in laryngeal squamous cell carcinoma and the presence of at least two as yet unidentified tumor suppressor genes in 9p13-23 region. Those putative tumor suppressor genes may become inactivated during the progression of the laryngeal squamous cell carcinoma. PMID- 12761948 TI - [Expression of vascular endothelial growth factor and nm23 as prognostic factors in nasopharyngeal carcinoma]. AB - OBJECTIVE: To evaluate the prognostic significance of vascular endothelial growth factor (VEGF) and nm23 expression for patients with nasopharyngeal carcinoma(NPC). METHODS: From Jan. 1991 to Dec. 1995, 75 NPC patients treated in Department of Radiotherapy & Chemotherapy, Zhongnan Hospital of Wuhan University, with follow-up of more than 4 yrs, were selected to enter this study. Specimens of 75 NPC were studied by immunohistochemical staining for VEGF and nm23. The immunohistochemical data were correlated with development of locoregional failure, distant metastasis and survival. RESULTS: Positive expression of VEGF and negative expression of nm23 were found in 54.7% and 56% of NPC specimens. Positive expression of VEGF and negative expression of nm23 correlated with the development of distant metastasis (P = 0.0181; P = 0.0091) and shorter survival (P = 0.0136; P = 0.0207). No association was observed with locoregional failure. No association was observed between the expression of VEGF and the expression of nm23.Cox proportional hazard model was used to compare the various factors and found the VEGF and nm23 expression were significant prognostic factors for patients with NPC. CONCLUSION: The results of the study show that the VEGF and nm23 expression are significant prognostic factors for patients with NPC and indicate that expression of VEGF and nm23 may be useful in identifying patients who may benefit from systemic chemotherapy or other novel treatment strategies. PMID- 12761949 TI - [Value of regional intra-arterial induction chemotherapy for locally advanced nasopharyngeal carcinoma]. AB - OBJECTIVE: To define the value of regional intra-arterial induction chemotherapy for locally advanced nasopharyngeal carcinoma (NPC). METHODS: 121 locally advanced NPC(stage III-IV) patients confirmed by pathology were randomly divided into two groups before radiation, and the two groups were given two different methods of chemotherapy: regional intra-arterial chemotherapy (IACT) and systemic chemotherapy (SCT). After radiation the effect of two methods was compared. RESULTS: The complete response rates were 27.9% in the IACT and 11.7% in the SCT, and 5 years survival rates were 63.9% in the IACT and 45.0% in the SCT respectively. The differences of two groups were significant. CONCLUSION: IACT is more reasonable than SCT as induction chemotherapy for these locally advanced NPC. PMID- 12761950 TI - [Surgery of substernal goiter]. AB - OBJECTIVE: To study the surgical approaches and operative techniques for substernal goiter. METHOD: A retrospective study of 27 cases with substernal thyroid nodules was made in our hospital. RESULT: The operations on 27 patients with substernal nodules have been successfully carried out. Among them, 10 were goiters, 9 adenomas, 4 malignancy and 4 were thyroid cancer metastasizing to paratracheal or substernal nodes. Resection via cervical collar incision was adequate in 23 cases. Three thyroid cancers with paratracheal node metastasis were completely resected by sternotomy and only one thyroid cancer with paratracheal and substernal nodes metastasis was resected by combined thoracotomy and cervical collar incision. CONCLUSION: Resection via cervical collar incision for all retrosternal thyroid nodules were advised. The preliminary experience with this procedure suggests that it has some advantages in the management of substernal goiter, including: 1. ease of operation compared to both sternotomy and thoracotomy; 2. relatively low morbidity; 3. safe and reliable. PMID- 12761951 TI - [A cell culture model in vitro for studying the permeability of cochlear microvascular endothelial cells in guinea pigs]. AB - OBJECTIVE: To establish an "in vitro" system for studying the permeability of cochlear microvascular endothelial cells. METHODS: Cochlear stria vascularis from guinea pigs were isolated by microsurgery, then cultured in vitro to get the monolayer cells of cochlear microvascular endothelium. The purity and property of cultured cells were identified by immunochemical method. The model in vitro was established by millcell technique of inoculation of the cultured monolayer cells of cochlear microvascular endothelium, then the permeability of this model was studied by 125I-bovine serum albumin. The positive group (brain microvascular endothelial cells), negative group (lung microvascular endothelial cells) and blank group (no cells) were conducted as control. RESULTS: 1. When cochlear stria vascularis fragment was isolated and cultured for two days, a few culture cells appeared around the fragments, then the amounts of culture cells increased. About ten days later, clusters of culture cells could be seen. Generally, spindle shaped single culture cells were observed after cloning. When subcultured for eight days, there formed a monolayer tightly packed against the neighbouring cells; these cells showed the cobblestone shape under the microscope. For immunochemistry, over 95% of the cultured cells showed a positive reaction to factor VIII related antigen. 2. The permeability of 125I-bovine serum albumin in testing group was higher than that in the positive group (P < 0.05) and was lower than that in the negative group and the blank group (P < 0.01). CONCLUSION: An ideal model in vitro has been established for studying the permeability of cochlear microvascular endothelial cells at molecular level. PMID- 12761952 TI - [The clinical characteristics of adhesive type cholesteatoma]. AB - OBJECTIVE: To study the clinical characteristics of adhesive type cholesteatoma. METHODS: Fifty-two cases (52 ears) of adhesive type cholesteatomas were reviewed. Of the 52 cases there were 30 males and 22 females; 24 left ears and 28 right ears. The age of the patients ranged from 8 to 70 years. All of the patients were treated by tympanoplasty, and the open and close methods were performed in 37 ears and 10 ears, respectively. Other methods were employed in the remaining 5 cases. Nine ears underwent tympanoplasty type "O" without ossicular re construction for an increase in the threshold of the bone conduction. Types I, III and IV tympanoplasties were performed in 1, 16 and 26 ears, respectively. RESULTS: The structures of the upper parts of the stapes had disappeared in 25 ears, which was significantly higher in this series than that in the attic type cholesteatoma. The over-all recovery rate of hearing was 69.8%, while 75.9% and 55.6% in the open and closed tympanoplasties, respectively. Postoperative complications of perforations and re-adhesions of the tense part of the tympanic membrane were observed in 3 and 4 ears, respectively. CONCLUSIONS: Adhesive type cholesteatoma is not uncommon. The cause of this disease may be the functional defects in the pharynotympanic tube. A thorough and effective treatment for the secretory otitis media is crucial for the prevention of adhesive type cholesteatomas. The open method tympanoplasty was considered as the first choice for adhesive type cholesteatoma while the closed method must be great careful. PMID- 12761953 TI - [Intact-bridge tympanomastoidectomy procedure: a reasonable option in the surgery of chronic otitis media and mastoiditis with cholesteatoma and/or granulation tissue]. AB - OBJECTIVE: To study and evaluate a surgical option for the treatment of otitis media and mastoiditis with cholesteatoma and/or granulation tissue. METHODS: The procedure is called intact-bridge tympanomastoidectomy (IBM). There are some modifications for the treatment of patients suffered from chronic otitis media and mastoiditis with cholesteatoma and/or granulation tissue. The features of IBM include: 1. the aditus, mastoid and attic opened and the intractable pathological tissues eradicated thoroughly. 2. the posterior tympanal space opened through the facial recess if necessary. 3. a widened middle ear space by lowering bony bridge established. 4. the adults blocked. 5. immediate or staging tympanoplasties conducted according to the conditions of mucosa in the tympanum. RESULTS: The modified IBM procedure was performed on 62 ears from 61 cases. The follow-up has been 12-38 months and more than two years in 51 ears. The dry ear was obtained within 5-13 weeks with an average of 6 weeks. In 24.2% of ears the air-bone gap(ABG) was 20dB HL or less and within 21-30 dB HL in 41.9% of ears, the ABG over 31 dB HL was 16.1%. CONCLUSION: Our results indicate that IBM procedure fulfills the desired purposes of both open- and close-cavity techniques, the thorough eradication of diseased tissue allow to get a dry ear and restoration of hearing. IBM, therefore, is a reasonable choice for the surgery of otitis media and mastoiditis with cholesteatoma and/or granulation tissue. PMID- 12761954 TI - [ACh and ATP-induced current in isolated outer hair cells and Deiters' cells of guinea pig cochlea]. AB - OBJECTIVE: To determine ACh- and ATP-induced currents in isolated outer hair cells of guinea pig cochlea, respectively. METHOD: The whole-cell patch-clamp technique was used to investigate potential and concentration dependence of ACh- and ATP-induced ion currents. RESULTS: ACh(100 mumol/L) induced an early inward current followed by a late outward current when the cells were voltage-clamped at -70 mV. Only outward current occurred when potential was over -60 mV and its reversal potential was (-80 +/- 5.61) mV(n = 6). When depolarization protocols were applied, amplitude of inward currents activated by ATP(100 mumol/L) decreased and reversed at (3.2 +/- 0.23) mV(n = 8). Ion currents activated by ATP (100 mumol/L) were also recorded from isolated Deiters' cells. CONCLUSION: ACh induced outward current was carried by K+ and ATP-induced inward current was through non-selective cations channel. Effects of ACh and ATP on OHCs were voltage dependent. PMID- 12761955 TI - [Rat hearing loss and hearing organs mitochondrial DNA4834 deletions associated with hypercholesteremia]. AB - OBJECTIVE: To determine whether or not the rat hypercholesteremia contributes to hearing organs mtDNA4834 deletion and involves in the development of hearing loss. METHODS: The rat hypercholesteremia model (n = 38) was established by feeding with high cholesterol diet and the control group (n = 22) with common diet for 6 months. The rats were tested for auditory sensitivity using auditory brainstem response (ABR). Twenty-one left cochleae and 27 left cochlear nuclei from experimental group and 10 left cochleae and 13 left cochlear nuclei from control group were harvested. The total DNA of them was extracted. mtDNA was amplified by nest PCR to examine the presence of mtDNA4834 deletion. RESULTS: Our result showed: (1) There is a significant increase in serum cholesterol level and ABR threshold in the experimental group. (2) The mitochondrially-encoded tRNA and ND1 segments were amplified from all samples, as well as mtDNA4834 deletions. (3) The incidence of mtDNA4834 deletions in hearing organs of hypercholesteremia rats was significantly higher than that of the control group (P < 0.05). CONCLUSION: Extended hypercholesteremia can induce hearing loss, and mtDNA4834 deletion in hearing organs may be one of the pathogenic mechanisms. PMID- 12761956 TI - [Combined interleukin-2 and herpes simplex virus thymidine kinase gene therapy for head and neck squamous cell carcinoma]. AB - OBJECTIVE: To assess the efficacy of combined interleukin-2 (IL-2) and herpes simplex virus thymidine kinase (HSV-TK) gene therapy for murine head and neck squamous cell carcinoma (HNSCC). METHODS: Ad IL-2 or/and Ad HSV-TK were injected into the tumor tissues directly after the murine HNSCC model was established. DL312 or PBS was used as control and ganciclovir (GCV) was used at 25 mg/kg for 7 days in Ad HSV-TK gene treatment groups. Tumor size was measured before and after treatment to evaluate the response to treatment. Cytotoxic T-lymphocyte (CTL) and natural killer (NK) assays were performed and IL-2 expressions were also measured after IL-2 gene transfection. RESULTS: HNSCC tumor growth was significantly inhibited following combined IL-2 and HSV-TK gene therapy as compared to other groups (P < 0.05). Increased levels of IL-2 protein expression was found in combined and single IL-2 treated groups. The combination and IL-2 treated groups produced greater activities of CTL and NK than that of the controls. CONCLUSION: IL-2 gene therapy can efficiently induce antitumor immunity of the host and enhance antitumor effects of HSV-TK. Combined IL-2 and HSV-TK gene therapy could significantly inhibit HNSCC tumor growth in the murine model. PMID- 12761957 TI - [Effect of transduction bax gene on experimental nasopharyngeal carcinoma]. AB - OBJECTIVE: To evaluate the effect of transduction bax gene on nasopharyngeal carcinoma (NPC). METHODS: The transduced bax gene was mediated by Dosper lipidsome to the HNE-1 cell lines, and confirmed by immunofluorescent stain. The apoptosis of the cell lines was studied by MTT stain and flow cytometry and the growth of implanted tumor in naked mouse after local injection of bax geneobserved. RESULTS: Instantaneous expression of bax gene in HNE1 cell lines was testified by immunofluorescent stain after transduction for 48 hours, the apoptotic index were 10.7% and 69.4% for pre- and post-transduction, respectively. MTT stain showed the values of A in 490 nm after transduction to be 2.004(36 h) and 1.902(48 h). The diameter of implanted tumor mass was (1.8 +/- 0.64) cm and (3.0 +/- 0.44) cm in experimental and control groups, respectively. CONCLUSION: The transduction bax gene can enhance the apoptosis of HNE1 cell lines and prevent growth of implanted tumor. PMID- 12761958 TI - [Speech discrimination with bilateral cochlear implants in noisy conditions]. AB - OBJECTIVE: To investigate the speech discrimination abilities of patients with bilateral cochlear implants. METHODS: The speech discrimination scores of Cantonese lexical tones were compared between two patients with unilateral and bilateral cochlear implants at different signal to noise (S/N) ratios. RESULTS: At favorable listening conditions with S/N ratios of +15, +10, and the mean binaural speech discrimination scores were 96%, 92% and 88%, but the mean monaural speech discrimination scores from the left and right ear were 86%, 83% and 74% respectively. At unfavorable listening conditions with adverse S/N ratios of 0, -5, -10 and -15, almost no speech discrimination scores could be obtained from monaural hearing but 80%, 72%, 68% and 54% respectively from binaural stimulation. CONCLUSION: Bilateral cochlear implants improve the discrimination of Cantonese lexical tones better than unilateral cochlear implant at various S/N ratios This study confirms the benefits of binaural hearing even in patients using bilateral cochlear implants. PMID- 12761959 TI - [Spontaneous otoacoustic emissions and efferent control of cochlea]. AB - OBJECTIVE: To study the relationship between spontaneous otoacoustic emissions(SOAE) and efferent control of cochlea and their clinical significance. METHODS: SOAE, transient evoked otoacoustic emissions (TEOAE), distortion product otoacoustic emissions (DPOAE) and contralateral white noise (60 dB SPL) suppression of TEOAE and DPOAE experiments were conducted in 312 ears of 95 patients with retrocochlear impairment and/or MOCS dysfunction and 64 normal young adults. RESULTS: MOCS dysfunction was shown in 126 ears of 65 patients (130 ears) with auditory neuropathy, 2 ears of 2 patients with unilateral acoustic neuroma, 4 ears of 2 patients with hyperacusis, 14 ears of 26 patients(48 ears) with normal hearing level in unilateral or bilateral tinnitus. Stronger EOAE could be recorded in total 146 ears with MOCS dysfunction at any pure tone hearing level. SOAE could be recorded in 126 of 146 ears (86.3%) with MOCS dysfunction and 44 of 128 ears (34.3%) with normal hearing. SOAE of ears with MOCS dysfunction was mainly at frequencies from 0.693 to 3.055 kHz and SOAE of normal ears was at frequencies from 1.135 to 2.746 kHz. Average value of maximum amplitude of SOAE spectrum (-3.4 +/- 6.4) dB SPL was significantly greater than that in normal ears (-6.8 +/- 7.8) dB SPL (P < 0.01). The major frequency range of SOAE (0.693-3.055 kHz) in MOCS dysfunction ears was essentially consistent with that of efferent suppression in normal ears (0.7-3 kHz). CONCLUSION: The modulation of the cochlear active mechanisms by MOCS mainly presents in the low- and mid-frequency regions, these frequencies correspond to the frequency range of SOAE. Stronger SOAE indicates pathophysiological significance. There is a clear clinical relationship between SOAE and the efferent modulation of the cochlea. PMID- 12761960 TI - [Influence of clinical staging of type III sinusitis on the operative effectiveness]. AB - OBJECTIVE: To explore the influences of different degree of pathological changes in chronic polypous sinusitis patients on the effectiveness of endoscopic sinus surgery. METHOD: According to history, anatomic structures and severity of lesions, 104 cases (208 sides) of Type III sinusitis were divided into 3 stages. Rates of clinical effectiveness and complications were compared and analyzed. RESULTS: The total effective rates were 93.06%, 82.89% and 70.00% and the rates of complications were 5.56%, 14.47% and 25.00% in stages 1, 2, 3 respectively. CONCLUSION: The effective rates, difficulties and risks of operation depend upon the anatomic variations (integration of ostiomeatal complex) and the degree of lesions (augmentation of granulation and osseous hypertrophy of ethmoid bone). All these should be listed as objective bases in classifying stages of patients which will play a directional role in clinical work. PMID- 12761961 TI - [Concentration of interleukin-5 and granulocyte-macrophage colony stimulation factor in nasal polyps tissue and its significance]. AB - OBJECTIVE: To study the relationship between the concentration of interleukin-5 (IL-5) and granulocyte-macrophage colony-stimulation factor (GM-CSF) in nasal polyps and its significance. METHODS: The concentration of IL-5, GM-CSF was measured by enzyme linked immunosorbent assay (ELISA) in a total of 38 cases, and the number of eosinophils were counted in the same tissues. RESULTS: The statistical analysis showed that the concentrations of IL-5 and GM-CSF have significant difference (P < 0.05) between the nasal polyps and the control group. There is a positive correlation between the quality of IL-5/GM-CSF and the extent of eosinophils (r = 0.75/0.71). CONCLUSION: This study demonstrated that there are inductive compartments, namely the microenvironment, in which immune-effector and inflammatory cell function is modulated in nasal polyps. A special discussion has been made of eosinophils and IL-5/GM-CSF in the formation of nasal polyps, and the effect of the IL-5/GM-CSF in the accumulation of eosinophils. PMID- 12761962 TI - [Clinical evaluation on second stage reconstruction for laryngostenosis after partial laryngectomy for laryngeal cancer patients]. AB - OBJECTIVE: To present a new method of decannulation for laryngostenosis patients after partial laryngectomy and extended partial laryngectomy, and restoration of the essential functions of larynx and normal neck appearance. METHODS: Nineteen cases of laryngostenosis after partial laryngectomy and extended partial laryngectomy were treated with second stage reconstruction. The patients had been treated by vertical laryngectomy (6/19) using extended vertical laryngectomy (11/19) and frontolateral laryngectomy (2/19). Among these patients, second stage reconstruction of larynx was performed by using rotary door myocutaneous flap (17/19). Stemohyoideus flap (1/19) and sternocleidomastoid flap (1/19). RESULTS: Three and five year-survival rates were 91.7% (11/12) and 3/5, respectively. Over all decannulation rate was 84.2% (16/19), but 94.1% (16/17) in patients with rotary door myocutaneous flap and 0% (0/2) in both patients with sternohyoideus flap and sternocleidomastoid flap. There were 3 decannulation failures (15.8%). All patients resumed acceptable voice, 94.7% (18/19) enjoyed satisfactory phonation, but 5.3% (1/19) showed severe hoarseness. All except 2 patients returned normal swallow function. The latter 2 patients experienced mild abnormal swallow during eating fluid food in early stage of surgery, but had normal swallow function after 1-2 weeks. CONCLUSION: Functional laryngectomy is a radical operation in selected cases with advanced laryngeal cancer. The second stage reconstruction with bi-pedical rotary door myocutaneous flaps can help decannulation in patients who developed laryngostenosis after partial laryngectomy and extended partial laryngectomy, and restore the essential function of larynx and normal neck appearance. PMID- 12761963 TI - [Surgical management of subglottic carcinoma]. AB - OBJECTIVE: To explore the surgical management and evaluate the effect on patients with subglottic cancer. METHODS: Fourteen cases with subglottic carcinoma were treated surgically from 1989 to 1998. There were T1-2N0 lesions in 5 cases, T3N0 2 lesions in 3 cases and T4N0-1 lesions in 6 cases. Eleven cases underwent partial laryngectomy and three cases underwent total laryngectomy. The defects of larynx were reconstructed by using uni-pedicled or bi-pedicled stemohyoid myofascial flap, platysma myocutaneous flap, sternocleidomastoid myoperiosteal flap, thyroid perichondral flap and epiglottis flap accordingly. Unilateral neck dissection was performed on 4 cases and bilateral on one. RESULTS: The function of phonation had been restored in all cases except in 3 patients who underwent total laryngectomy. Seven out eleven(63.6%) were decannulated. The swallowing function was restored in all patients. In all cases the 3 and 5 year survival rates were 78.6%(11/14) and 63.6% (7/11), respectively. CONCLUSIONS: The conservative surgery can be used for the majority of the subglottic carcinoma if the lesions were entirely resected. The proficiency in surgical methods is crucial for the preservation of laryngeal function. PMID- 12761964 TI - [Study on vibration mode of different phonatory source and compensation after partial laryngectomy]. AB - OBJECTIVE: To investigate the nature of pathological voice production and compensatory mechanism after partial laryngectomy. METHODS: Vocal function of 93 cases following partial laryngectomy (28 cases of horizontal partial laryngectomy, 35 vertical partial laryngectomy, 30 horizontal-vertical partial laryngectomy) were examined by acoustic analysis, aerodynamic analysis and videostroboscopic examination. RESULTS: Horizontal supraglottic laryngectomy(HL) resulted in slight dysphonia after operation. The cover of vocal fold was hypertrophic and edematous. Mucosa wave was increased. Acoustic analysis showed significant difference between normal and horizontal laryngectomy groups(P < 0.05). Vertical laryngectomy(VL) resulted in moderate-severe dysphonia. Acoustic analysis was significantly worse in VL than in normal group(P < 0.01). The nonglottic phonatory source was the contralateral ventricular fold or contralateral vestibular mucosa (arytenoid mucosa, root mucosa of glottis) to approximate the reconstructed flap. It was noted that contralateral vocal fold did not participate in vibration. Horizontal-vertical partial laryngectomy (HVL) resulted in moderate-severe dysphonia. Acoustic analysis of HVL was significantly worse than that of normal phonation(P < 0.01). The site of compensatory mucosa vibration upon phonation was the contralateral vocal fold or contralateral hypertrophic arytenoid mucosa. The thinner the flap, the better the vocal quality was. Vocal quality of VL was worse than that of HVL in regard to shimmer, jitter, normalized noise energy, maximal phonation time and harmonic-noise ratio, but there was no significant difference between them. CONCLUSIONS: Partial laryngectomy has different vibrating compensatory modes: Voice function of horizontal partial laryngectomy was the best as it preserved the normal vibration mode; vertical partial laryngectomy was the worst with the nonglottic vibrating source of reconstructed flap-ventricular fold. The reconstructed flap, ventricular fold, mucosa of epiglottis and arytenoid take predominantly part in neoglottal vibration. The situation, volume and pliability characteristic of reconstructed flap was also important to vocal quality. PMID- 12761965 TI - [Clinical study of juvenile-onset recurrent respiratory papillomatosis]. AB - OBJECTIVE: To study the clinical behavior of juvenile-onset recurrent respiratory papillomatosis in order to find some factors correlated to the development of this disease, and to sum up the significance and experience of CO2 laser surgery. METHOD: Sixty patients with juvenile-onset recurrent respiratory papillomatosis from September 1995 to December 1998 were retrospectively analyzed. RESULTS: The age of onset in 50 cases (83.3%) was below 4 years, and the peak-age was 2 years. The rates of recurrence were 72.0% and 45.7% (chi 2 = 4.71, P < 0.05) below and over 2 years, respectively. The rates of aggressive disease were 88.0% and 54.3% (chi 2 = 7.66, P < 0.01) below and over 2 years, respectively. The predominant sites of the disease were the vocal cords, the false vocal cords, the laryngeal ventricle, the laryngeal surface of the epiglottis and the subglottic region. Tracheostomy induced the development of tracheal papilloma, therefore should be avoided as possible. Laryngeal papilloma might be divided into four types on the basis of the growth manner and surface form corresponding to clinical behaviors. Five patients were followed-up for 1.5 years without recurrence, 18 patients had fewer recurrences following treatment, 33 patients were under treatment, and 3 patients died. Nineteen patients lost follow-up. The major complications included laryngeal and tracheal stenosis. CONCLUSION: Clinical behaviors of juvenile-onset recurrent respiratory papillomatosis were relevant to the age, growth form and tracheotomy. CO2 laser was an ideal instrument for ablation of the laryngeal papillomas with the following advantages: simple management, less bleeding, preservation of laryngeal structure and avoidance of tracheostomy. PMID- 12761966 TI - [Important prognostic factors in patients with skull base erosion from nasopharyngeal carcinoma after radiotherapy]. AB - OBJECTIVE: To evaluate the long-term outcome and prognostic factors in patients with skull base erosion from nasopharyngeal carcinoma (NPC) after initial radiation therapy. METHODS: From January 1985 to December 1986, 100 patients (71 male, 29 female) with the diagnosis of NPC were found to have skull base erosion from computed tomography (CT). The mean age was 41 (16-66) years. Ninety-six patients had World Health Organization (WHO) type III undifferentiated carcinoma while four had type I carcinomas. Metastatic workup including chest radiography, liver ultrasound and liver function test were negative. All patients underwent external beam radiotherapy alone to 66-80 Gy over 6-8 weeks. Daily fraction size of 2 Gy was delivered utilizing cobalt-60 or linear accelerator. No patient received chemotherapy. All patients were followed up at regular intervals after irradiation. Median follow-up was 22.3 months (2 months-174 months). Overall survival of the cohort was computed by the Kaplan-Meier method. Potential prognostic factors on survival were examined. Multivariate analyses was performed using Cox regression model. RESULTS: One, three, five, and ten year overall survival rates for the cohort were 79%, 38%, 27% and 13%, respectively. However, in the sub-group of patients with both anterior cranial nerves (I-VIII) and posterior cranial nerve (IX-XII) involvement had a five-year survival of only 7.7%. Causes of death included local recurrence (59 patients), distant metastases (21 patients), both local recurrence and distant metastases (1 patient) and unrelated causes (5 patients). After multivariate analysis, complete recovery of cranial nerve involvement, cranial nerves palsy and recovery of headache after irradiation were found to be independent prognostic factors in this cohort. CONCLUSION: This report presents one of the longest follow-ups of patients with nasopharyngeal carcinoma invading skill base. It demonstrates the importance of cranial nerves involvement as well as recovery of headache and cranial nerve palsy. These factors should be evaluated carefully from history and physical examination as well as imaging studies. Currently available magnetic resonance imaging (MRI) is highly recommended as an additional test. A subgroup of patients with skull base involvement had long-term survival after radiotherapy alone. More aggressive strategy such as combined chemo-radiotherapy and altered fractionation radiotherapy may improve outcome in patients with poor prognostic factors. PMID- 12761967 TI - [Expression and significance of serum soluble tumor necrosis factor receptor-I in patients with head and neck neoplasm]. AB - OBJECTIVE: To investigate the expression and significance of serum soluble tumor necrosis factor receptor-I (sTNFR-I) in patients with head and neck neoplasms. METHOD: The study was undertaken to detect serum sTNFR-I levels in 160 head neck cancer patients (including 12 cases of malignant lymphoma, 62 nasopharyngeal cancer, 56 laryngeal cancer, 22 hypopharyngeal cancer, 3 maxillary cancer and 5 thyrophyma), using Sandwich enzyme-linked immuno-sorbent assay. RESULTS: The sTNFR-I levels were significantly higher in patients with carcinoma than those of healthy controls (P < 0.01). The sTNFR-I levels of malignant lymphoma and nasopharyngeal carcinoma were the highest among patients with head and neck neoplasm. The correlations between the serum sTNFR-I levels and the stages of head and neck neoplasms and other laboratory parameters were also analyzed and discussed. CONCLUSION: The serum sTNFR-I levels may reflect the human immunity function, therefore it can be used as a helpful indicator to evaluate the therapeutic effect and monitor the relapses and metastasis of cancer. PMID- 12761968 TI - [Expression of vascular endothelial growth factor and transforming growth factor beta 1 in nasal polyps]. AB - OBJECTIVE: To study the expression and significance of vascular endothelial growth factor/vascular permeability factor (VEGF/VPF) and transforming growth factor-beta 1(TGF-beta 1) in nasal polyps. METHODS: Expression of VEGF/VPF and TGF-beta 1 in nasal polyps from 34 patients and middle turbinates from 30 patients with deviation of nasal septum was prospectively studied with immunohistochemistry. Tissue sections were observed under optical microscope. RESULTS: (1) The VEGF/VPF positivity in vascular endothelium and in glandular cell was significantly higher in nasal polyps than in middle turbinates (P < 0.01 and P < 0.05, respectively); (2) The TGF-beta 1 positivity in extracellular matrix and in cells in the stroma was significantly higher in nasal polyps than in middle turbinates(P < 0.005); (3) The distribution and shape of TGF-beta 1 expressing cells in nasal polyps were similar to that of eosinophil, their positivities were significantly correlative; (4) The positivity of VEGF/VPF and TGF-beta 1 did not correlate with clinical type in nasal polyps (P > 0.05). CONCLUSION: (1) The VEGF/VPF may play a key role in the formation of heavy edema of nasal polyps; (2) The TGF-beta 1 may contribute to some of the pathologic changes observed in nasal polyps, such as thickening of the epithelial basement membrane and stromal fibrosis; (3) Eosinophils in nasal polyps represent a major source of TGF-beta 1. PMID- 12761969 TI - [Expressions of vascular endothelial growth factor and basic fibroblast growth factor in nasal polyp and its role]. AB - OBJECTIVE: To evaluate the role of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor(bFGF) in the pathophysiology of nasal polyp. METHODS: Thirty-nine patients with nasal polyp were divided into two groups: group A(representing type 1 and type 2 phase 1-2) and group B (representing type 2 phase 3 and type 3). The expression of VEGF and and bFGF in both groups were studied with immunohistochemical method. RESULTS: VEGF and bFGF were not detected in norms. The detection rates of VEGF and bFGF were 59%, 41%, and 71%, 80% in group A and group B respectively. The positive rate and the number of positive cells were higher significantly in group B than that in group A. VEGF and bFGF were located mainly in the inflammatory cells and epithelial cells around the basilar membrane and inflammatory cells and endothelial cells around the vessel. CONCLUSIONS: The overexpression of VEGF and bFGF in nasal polyp may contribute to the growth of vessels, accumulation of inflammatory cells, as a result, to enhance the development of nasal polyposis. This phenomenon maybe an important histological mark distinguishing ordinary polyp from polyposis. PMID- 12761971 TI - [Expression and significance of interleukin-6,8 in human nasal polyps]. AB - OBJECTIVE: To explore the role of interleukin-6,8 in nasal polyp formation and to search into the effect of allergy in the pathogenesis of nasal polyps (NP). METHODS: The expression and significance of interleukin-6,8 were studied in 36 nasal polyps and 36 serum samples of NP patients by radioimmunoassay. RESULTS: The mean value of IL-6 and IL-8 was (2.7658 +/- 0.3797) ng/L and (4.1877 +/- 0.1758) ng/L in all nasal polyp tissue homogenates. As compared with serum of NP patients, IL-6 and IL-8 were over expressed in nasal polyp tissue homogenates. No relation was found between the expression of IL-6/IL-8 and patients' gender, age and clinical stage. The expression of IL-6 and IL-8 in patients' serum, cord blood and normal serum showed no significant difference. CONCLUSION: IL-6 and IL 8 are strongly correlated with the formation of nasal polyp. Neither allergy nor infection play a role in the pathogenesis of nasal polyps. PMID- 12761970 TI - [The protein expression of Bcl-x1 and Bcl-2 in eosinophils in nasal polyps and the influence of beclomethasone dipropionate on the expression]. AB - OBJECTIVE: To investigate the protein expression of B-cell lymphoma/leukemia 2(Bcl-2) and B-cell lymphoma/leukemia-x long(Bcl-x1) in eosinophils in nasal polyps and the influence of beclomethasone dipropionate on the expression. METHODS: Using May-Grunwald-Giemsa (MGG) method and immunohistochemical method, the protein expression of Bcl-x1 and Bcl-2 in eosinophils in nasal polyps from patients treated with beclomethasone dipropionate treatment and patients without any treatment was compared. RESULTS: (1) Nasal polyp tissues from patients without treatment had more eosinophils than those from patients with treatment(P < 0.01). (2) No protein expression of Bcl-2 was observed in all 52 patients. (3) 20.0% patients with treatment had the expression of Bcl-x1, whereas 48.1% patients without treatment had the expression. The difference between these two groups was significant(P < 0.05). CONCLUSION: These data suggest that Bcl-x1 may act as an anti-apoptotic molecule in eosinophils and corticosteroids induce eosinophil apoptosis through suppressing the expression of Bcl-x1. PMID- 12761972 TI - [The experimental study of intratemporal facial nerve regeneration in chitin chamber]. AB - OBJECTIVE: To investigate the feasibility of intratemporal facial nerve regeneration in a chitin chamber. METHODS: A left 6 mm intratemporal facial nerve gap was created in adult white rabbit, and a total of 48 animals were used. The proximal and distal stumps were bridged with chitin chamber (experimental group) and silicone chamber (control group). Regeneration of the nerve was assessed by gross observation, nerve electrophysiological test, histological examination and digital morphological analysis in both groups. RESULTS: 1. Three months after implantation, the proximal transected neural stump was connected with the distal stump in the chitin chamber. Five months after operation, the regenerated nerve exhibited a more nature fascicular organization and the chitin tubes were completely absorbed. 2. Nerve evoked potentials showed better conduction ability in the chitin chamber. There were no significant difference in electrophysiological indices between the two chamber groups. 3. Most of the regenerated nerve fibers were myelinated nerve fibers, and axons and myelins had similar histograms between the experimental and control groups. 4. Five months after operation, the mean percentage of recovery of the regenerated nerve fiber area, axonal area and axonal number were more than 71% in the experimental group. There was no significant difference between the control and experimental groups. CONCLUSION: These results suggested that intratemporal facial nerve could regenerate effectively within the chitin chamber in rabbits and such kind of chamber may be clinically useful. PMID- 12761973 TI - [Glutamate induced modulation of free Ca2+ in isolated inner hair cells of the guinea pig cochlea]. AB - OBJECTIVE: To investigate the modulation effect of glutamate (Glu) on intracellular free Ca2+ concentration([Ca2+]i) of inner hair cell (IHC). METHODS: Using the laser scanning confocal microscope (LSCM), the exogenous Glu-induced changes in [Ca2+]i of isolated 10 IHCs and 10 OHCs of guinea pig cochlea were observed with fluo-3, a fluorescent probe for [Ca2+]i. RESULTS: The IHCs were identified by their unique flask shape with a distinct neck and spherical base and a large spherical nucleus. In most cases, normal cell shapes could be maintained about two hours after isolation. The images of [Ca2+]i from LSCM were similar to those from inverted microscopy. Fluorescence of Fluo-3 distributed in the isolated IHCs with brighter staining in the nucleus. In the presence of low concentration of Glu (3.85 mumol/L), there was an increase of [Ca2+]i in IHCs, whereas no change in OHCs was found. Of the 10 IHCs, increases of [Ca2+]i were observed in 9 and no change in 1. Of the 10 OHCs, 7 showed no [Ca2+]i change and only 3 showed minor reduction of [Ca2+]i. An increase of the Glu concentration (21.88 mumol/L) induced a corresponding increase of [Ca2+]i in IHCs, but eventually resulted in a gradual decrease of [Ca2+]i with a distortion of the normal shape, which indicated that the IHCs were degenerated and swelling. CONCLUSION: These results suggested that exogenous Glu is capable of modulating [Ca2+]i of IHC and may be act on autoreceptor in a positive feedback manner. Excessive Glu induced the accumulation of IHC [Ca2+]i which finally resulted in the degeneration and edema of IHC and the reduction of IHC [Ca2+]i. PMID- 12761974 TI - [The effects of streptomycin on the calcium influx in SGCs during depolarization]. AB - OBJECTIVE: To examine the effect of streptomycin on the calcium influx in spiral ganglion cells (SGCs) during depolarization and the differences among different concentrations of streptomycin and SGCs sources with the aim of exploring the mechanism of acute streptomycin ototoxicity. METHODS: The SGCs of guinea pig cochlea were isolated using an enzyme-machine method and loaded with 10 mumol/L Fluo-3/AM for 30 min at 37 degrees C. Dissociated SGCs loaded with Fluo-3 were examined with a confocal microscope (ACAS Ultima, USA) using a 20 x objective lens and linear scan mean. The fluorescent images collected every 5-sec for a total of 300 sec were stored in a computer. The fluorescent intensity of the images was analyzed by a software cooperated with the confocal microscope, and a curve of fluorescent intensity changes against time was obtained. RESULTS: The [Ca2+]i of SGCs was steady under the perfusion with standard extracellular solution. The [Ca2+]i of SGCs increased in 12/14 cells under the perfusion with 150 mmol/L high potassium solution, but increased in only 1/10 cells induced by the high potassium calcium free solution. After treating with 1 mmol/L, 0.1 mmol/L, 0.01 mmol/L streptomycin, the [Ca2+]i of the SGCs perfused with the potassium solution was increased in 0/9, 6/14, 7/13 cells, respectively. After the SGCs were treatment with 0.02 mmol/L streptomycin, the [Ca2+]i of the SGCs perfused with the high potassium solution was increased in 5/10 SGCs from the apical turn and in 0/15 SGCs from the basal turn. CONCLUSION: Perfusion with high potassium media can result in obviously increased [Ca2+]i of the SGCs, and the increase of the [Ca2+]i in the SGCs may originate from the extracellular calcium influx. The calcium influx can be blocked by streptomycin and such effects depend on streptomycin concentrations and SGCs sources. PMID- 12761975 TI - [Expression of alpha and beta subunit isoforms of Na, K-ATPase in the guinea pig endolymphatic sac]. AB - OBJECTIVE: To investigate the expression and significance of alpha and beta subunit isoforms of Na, K-ATPase in the guinea pig endolymphatic sac. METHODS: The distribution of alpha and beta subunit isoforms of Na, K-ATPase in endolymphatic sac of guinea pig was identified by immunocytochemistry and in situ hybridization. RESULTS: The expression of Na, K-ATPase alpha and beta subunit isoforms varied among different cell regions of the endolymphatic sac. Epithelial cells of the endolymphatic sac were observed to contain the alpha 1, beta 1 and beta 2 subunit isoforms, and the expression of beta 2 was stronger than that of beta 1. Subepithelial cells contained alpha 1 and alpha 2 subunit isoforms and the expression of alpha 1 was stronger than that of alpha 2. No expression of alpha 3 subunit isoform was observed in endolymphatic sac. CONCLUSION: Na, K ATPase of endolymphatic sac consists of different alpha and beta subunit isoforms which, working in concert, serve to maintain homeostasis in inner ear. PMID- 12761976 TI - [Expression of BDNF and FGF-2 following axotomy in rat facial motoneurons]. AB - OBJECTIVE: To study the expression and distribution of brain derived neurotrophic factor (BDNF) and fibroblast growth factor-2 (FGF-2) in normal facial motoneurons (FMNs) and in FMNs following axotomy. METHODS: The right facial nerves were transected 6 mm distal to the stylomastoid foraman in adult Wistar rats except the normal group. Serial 20 microns cryosections were cut through the whole brainstems. Expressions of BDNF and FGF-2 mRNA as well as BDNF and FGF-2 protein were studied by in situ hybridization, immunohistochemistry and image analysis. RESULTS: BDNF mRNA and its protein were observed in widespread areas of normal rat facial nucleus, and those increased 1 day after axotomy. In addition to neurons, glial cells were also stained. FGF-2 mRNA and its protein were mainly localized in normal FMNs of ventral facial nucleus. After axotomy, expression of FGF-2 mRNA started to up-regulate in FMNs at 7 days, however FGF-2 protein drastically reduced at 3 and 7 days. CONCLUSION: In addition to target supporting, there may be BDNF autocrine and paracrine mechanisms as well as FGF-2 autocrine mechanisms in normal rat FMN. When target-derived BDNF is deprived, alternative sources of BDNF support may substitute immediately after axotomy. However, deprivating of target--derived FGF-2 may result in down-regulation of FGF-2 protein in adult rat FMNs at early stage after axotomy. These suggest that the exogenous FGF-2 might provide a supportive environment for the recovery of metabolism and function of FMNs at early stage following axotomy. PMID- 12761977 TI - [Observation of facial nerve evoked potentials in rabbits]. AB - OBJECTIVE: To investigate the possibility of recording the Antidromic Nerve Evoked Potentials (ANEP) from rabbit's scalp. METHODS: With electrical stimulation of peripheral segment of facial nerve outside the stylomastoid foramen in rabbits, recordings were made on scalp using low-impedance silver disc electrode. RESULTS: At the vertex, clear ANEPs composed of two waves, wave P1 and P0, were recorded in all experimental animals. CONCLUSION: The study shows that ANEPs can be recorded on rabbits' scalp. The ANEPs have two waves, P1 and P0. The sources of the responses are from facial nerve and its muscle respectively. PMID- 12761978 TI - [Expression of matrix metalloproteinasa-2,9 in cholesteatoma and middle ear cancer]. AB - OBJECTIVE: To study various expressions and roles of MMP-2, 9 in cholesteatoma and middle ear cancer. METHODS: MMP-2 and MMP-9 were detected with immunohistochemical methods using monoclonal antibody in 36 cholesteatomas, 10 CAMS and 16 middle ear cancer. RESULTS: The stain in cholesteatoma was stronger than that in CAMS and NAMS. Expressions of MMP-2, 9 were seen in all layers of the epithelium in CA and the stain was strongest in basement membrane. The positive rates of stain was 83% and 50% in CA and CAMS respectively. There was weak stain in 2 NAMS. In middle ear cancer, the expression rates of MMP-9 and MMP 2 were 50% (8/16) and 56.25% (9/16) respectively. The expression level was correlated with tumor differentiation and its clinical stage. Expressions of MMP 2 and MMP-9 were closely related in cholesteatoma. There was no relationship between expressions of MMP-2 and MMP-9 in middle ear cancer. CONCLUSION: Cholesteatoma has a similar feature with middle ear cancer in invading surrounding of surround bone matrix. The disorder between MMPs and its inhibitor was one of reasons for bone absorption of cholesteatoma and middle ear cancer. PMID- 12761979 TI - [Research of the chorda tympani nerve in cholesteatoma]. AB - OBJECTIVE: To investigate the ultrastructure of the chorda tympani nerve and analyze the taste and facial nerve functions in patients with cholesteatoma. METHODS: 1. The tympanic segments of chorda tympani nerves were collected for ultrastructural investigations in 19 cholesteatoma cases who underwent canal-wall down tympanoplasty. 2. All these patients received a spatial (regional) taste test preoperatively and postoperatively, respectively. Multiple loci were tested in a given order (front of the tongue, foliate papillae, circumvallate papillae, and soft palate). The solutions used in this study were 1.0 mol/L sodium chloride (salty), 1.0 mol/L sucrose (sweet), 0.032 mol/L citric acid (sour) and 0.001 mol/L quinine hydrochloride (bitter). The analyses of variance was used. 3. House Brackmann facial nerve grading system was conducted to evaluate the facial nerve function preoperatively and postoperatively. RESULTS: 1. There were obvious ultrastructural damages in all the chorda tympani nerves, such as swelling (100%), disarrangement (100%), vacuoles formation of myelin (89%), edema of Schwann cells (95%), intracytoplasmic vacuoles in Schwann cell (89%) and proliferation of the collagen tissue (89%). 2. Two patients complained of change of taste after operation. The analyses of variance showed that the taste function of the ipsilateral side of tympanoplasty were not statistically altered for each stimulus at each locus (P > 0.05). 3. No facial palsy occurred postoperatively. CONCLUSION: This research suggested that the chorda tympani nerves underwent ultrastructural changes in patients with cholesteatoma. The dissection of chorda tympani nerve would not affect the taste and facial nerve functions. PMID- 12761980 TI - [Application of an image-guidance system in endoscopic sinus surgery]. AB - OBJECTIVE: To evaluate the advantages of image-guidance system in endoscopic sinus surgery. METHODS: Twenty-eight endoscopic sinus surgery procedures were performed with the help of image-guidance system. There were 11 cases of chronic sinusitis with/or without nasal polyps, 4 juvenile nasopharyngeal angiofibromas, 6 pituitary adenomas, 3 ethmoid ossifying fibromas, 2 nasopharyngeal mixed tumors, 1 nasal leiomyoma, 1 fungal sinusitis, and 1 inverted papilloma. RESULTS: In all cases, the preoperative operation would take 15-30 minutes, the registration rate was 1.3-2.0, the localization accuracy was within 1 mm. Compared with the traditional endoscopic sinus surgery, the operative time did not differ obviously. No complication occurred. CONCLUSIONS: The image-guidance system could identify the operative borders and critical anatomical structures together with the corresponding CT data, especially in cases with anatomical variations, unidentified anatomical landmarks, and intranasal or anterior skull base tumors. Endoscopic sinus surgery, combined with the image-guidance system, provided accurate tumor resection while preserving normal tissue. It could increase surgical effectiveness and decrease overall surgical complications. We believe that the image-guidance system is a useful tool for endoscopic sinus surgery. PMID- 12761981 TI - [Maxillofacial vascular malformation associated with abnormal communication between external carotid and cranial arteries]. AB - OBJECTIVE: To introduce the clinical characteristics of maxillofacial vascular malformation with abnormal communication between external carotid and cranial arteries. METHODS: One hundred and twenty patients with maxillofacial vascular malformations had been studied by arteriography of internal and external carotid, and vertebral arteries before embolization of tumor supplying artery. Cases found to have communications between extra- and intra-cranial arteries were analyzed. RESULTS: Fourteen patients (11.67%, 14/120) were found to have abnormal communications between external carotid and cranial arteries. Among them, 11 patients demonstrated communications between occipital and vertebral arteries, 1 patient showed ascending pharyngeal artery and vertebral artery communication, and 2 patients showed maxillary artery-ophthalmic artery communication. CONCLUSION: Embolization of tumor supplying artery is a safe and practical method for the treatment of maxillofacial vascular malformation when done under digital subtraction angiography and superselective catheterization to avoid the abnormal communicant branches. PMID- 12761982 TI - [Differences of anatomic variations in ostiomeatal complex between two sides of the deviated septum]. AB - OBJECTIVE: To explore the influence of the deviated septum on the development of ostiomeatal complex (OMC). METHODS: Nasal endoscopic examination and CT scan of the paranasal sinuses of 103 patients with nasal septal deviation were analyzed. The differences of anatomic variations in OMC between ipsilateral and contralateral sides in relation to the direction of septal deviation. The incidence of sinusitis on either side of the deviated septum was evaluated. RESULTS: On the side opposite to the deviated septum, the incidence of middle and inferior turbinate hypertrophy was higher than that of the deviated side(P < 0.05). The width of agger nasi and ethmoid bulla was on the contralateral side larger than that of the ipsilateral side (P < 0.01). The incidence of paradoxical curvature of middle turbinate on the ipsilateral side was higher than that of contralateral side (P < 0.01). Other anatomic variations showed no significant differences between two sides of the deviated septum (P > 0.05), nor was there any difference between the incidence of sinusitis on the two sides. CONCLUSION: Some compensatory changes in middle and inferior turbinates and lateral nasal wall on the opposite side to the deviation might be caused by the deviated septum. PMID- 12761983 TI - [Effect of p14ARF gene on cell growth of human laryngeal tumor cells and expression of endogenous p53 protein]. AB - OBJECTIVE: To explore the inhibitory effect of p14ARF on the cell growth of laryngeal carcinoma and the expression on endogenous p53. METHODS: p14ARF cDNA was transferred to the cell line Hep-2 of squamous cell carcinoma of the larynx by gene transfer to study the cell cycles and the expression of endogenous wild type p53 using flow cytometry, polymerase chain reaction(PCR) and Western Blotting. RESULTS: Expression of p14ARF significantly affected the Hep-2 cell growth. The clone-forming efficiency of the Hep-2 cells transferred with p14ARF was 57%, compared with empty vector pcDNA3. The number of 48 hours after transfer with p14ARF cDNA at both G0/G1 and G2/M was two fold as the control. The expression of endogenous wild type p53 was significantly enhanced. CONCLUSION: Expression of p14ARF can up-regulate the expression of endogenous wild type p53 and inhibit the Hep-2 cell growth of human laryngeal squamous cell carcinoma at both G0/G1 and G2/M. PMID- 12761984 TI - [Study of Newcastle disease virus in the treatment of human laryngeal squamous carcinoma]. AB - OBJECTIVE: To explore the biological value of newcastle disease virus in the therapy of human laryngocarcinoma. METHODS: Nude mice model bearing laryngocarcinoma were established using human laryngeal squamous carcinoma cell line (Hep-2). Large amounts of Newcastle disease virus(NDV) were injected into the tumor. Changes in carcinoma administrated with the NDV were observed under light and electron microscopes. Isolation of NDV was attempted and sera were examined using the method of enzyme-linked immunosorbent assay (ELISA). RESULTS: The difference between experimental and control groups was statistically significant, including the average weight of the mice: t' = 2.397 (P < 0.05), the average weight of the tumor tissue: t' = 2.852 (P < 0.05) and the average volume of the tumor tissue: t' = 6.058 (P < 0.01). In the experimental group, the NDV was isolated in the tumor tissue and ELISA was positive. The necrosis of the tumor cell and inflammatory cellular infiltration were found under light and electron microscopes. CONCLUSION: These results indicated that NDV was effective in the treatment of laryngocarcinoma and had no damage to normal tissue. PMID- 12761985 TI - [Detection of mutations in exon 7 of p53 from fresh tumor samples by single strand conformation polymorphism and direct DNA sequencing]. AB - OBJECTIVE: To detect mutations of exon 7 of p53 gene in laryngeal squamous cell carcinoma (LSCC) and analyze the relationship between the mutation and LSCC. METHODS: Fresh tumor samples from 60 LSCC patients were examined using polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) and direct DNA sequencing. RESULTS: An extra band in SSCP 3 was shown in 25 out of 60 samples. Exon 7 of p53 were detected by direct DNA sequencing in 8 samples taken at random from 25 cases. In 8 cases, 1 base pair deletion CCC-->CC in 250 codon which may result in frame shift mutation was shown. Simultaneously, in 3 cases, transversion AGT-->GGG in 261 codon which may bring out missense mutation was found. In 2 cases, 1 base pair deletion CGG-->GG in 248 codon was detected. In another case, 1 base pair deletion AGG-->AG in 249 codon was detected. Transversion ATC-->TTC in 254 codon was found in one of the cases. Transversion GGT-->TCA in 262 codon was found in another case. CONCLUSION: The codon 248, 250, 261 may be "hot spots" in LSCC. Mutations and deletions in exon 7 of p53 gene occurred frequently in LSCC. PMID- 12761986 TI - [Clinical study of the sentinel lymph node of patients with laryngeal and hypopharyngeal carcinomas]. AB - OBJECTIVE: To investigate the methods of detecting the sentinel lymph node of laryngeal and hypopharyngeal carcinomas and its predictive value in the cervical metastasis of the carcinoma. METHODS: In 29 patients who suffered from laryngeal or hypopharyngeal carcinoma with NO neck, the patent blue was injected into the surrounding tissue of the tumor during the operation to identify the sentinel lymph nodes. The sentinel lymph nodes were dyed blue. The frozen histopathology was done during the operation, the lymph nodes at the ipsilateral side of the neck were dissected completely, and the routine histopathology was done as the gold standard to study the predictive value of the sentinel lymph node in the metastasis of the cervical node. RESULTS: Of 29 patients, 28 patients' sentinel lymph nodes were detected successfully during operation. There was an average of 2.5 lymph nodes per side per patient. Most of the sentinel lymph nodes were in the level II and level III regions of the ipsilateral side of the neck, and there were bilateral sentinel nodes in patients suffered from superglottic carcinoma. Three patients' sentinel lymph nodes were found to be positive in the frozen inspection, and the routine histopathology confirmed the result. The micrometastasis rate was 10.7% (3/28). There were no metastatic lymph nodes found in patients who were negative for the sentinel lymph nodes during the operative frozen histopathology. The predicted value of the sentinel lymph nodes to the cervical lymph node metastasis was 100 per cent. CONCLUSION: There is a very important predicted value of sentinel lymph nodes in the cervical metastasis of patients suffered from laryngeal and hypopharyngeal carcinomas. It could reduce the neck dissection in patients with laryngeal and hypopharyngeal carcinomas. PMID- 12761987 TI - [Detection mucin gene in cervical lymph nodes in supraglottic cancer by reverse transcription polymerase chain reaction]. AB - OBJECTIVE: To investigate the efficacy of discovery of cervical lymph nodes micrometastasis of supraglottic carcinomas by detection of mucin (MUC1) gene. METHODS: Twenty-five supraglottic carcinoma patients with clinically negative lymph nodes were performed neck dissection. MUC1 gene mRNA in these resected lymph nodes was detected by reverse transcription polymerase chain reaction (RT PCR) and histopathological examination to find the potential micrometastasis. RESULTS: Of all 93 cervical lymph nodes, micrometastasis were found in 23 nodes of 6 case by histopathological examination and RT-PCR. Four of 19 cases with negative lymph nodes pathologically showed MUC1 gene expression and were identified to have metastasis by clinical recurrence and serial-sectioning. Therefore, the rate of detection of micrometastasis by this way was 16% (4/25), higher than that of regular pathologic methods. CONCLUSION: Detection of lymph nodes micrometastasis with RT-PCR method was more sensitive than that of regular pathologic methods. It is a valuable method to detect early lymph nodes metastasis. PMID- 12761988 TI - [Value of surgical margin in directing the treatment of early laryngeal carcinoma after partial laryngectomy]. AB - OBJECTIVE: To investigate the value of surgical margin in directing the treatment of early laryngeal carcinoma after partial laryngectomy. METHODS: We studied 87 cases of T1N0M0 and T2N0M0 laryngeal carcinoma in which the surgical margins were reserved during partial laryngectomy. The recurrence and survival rates between the positive and negative margin groups were compared; the survival between the positive margin groups with postoperative radiotherapy and negative groups; the survival between groups with and without postoperative radiotherapy in negative margins. RESULTS: Recurrent rate of the negative group was lower than that of the positive group (5.8% vs 33.3%, chi 2 = 10.64 P = 0.001). The estimated survival was higher in the negative group than in the positive group(P = 0.011), and also in tumor-free time(P = 0.007). The survival of those with positive surgical margin and received postoperative RT was lower than that with negative margins(P = 0.01). In the negative group, the difference in the survival between those with or without postoperative RT was not significant (P = 0.405). CONCLUSION: The prognosis of the positive margin group of the early laryngeal carcinoma was worse than that of the negative group, but postoperative RT can improve it; it is not necessary for the negative group to be treated by postoperative RT. PMID- 12761989 TI - [Preservative surgery of medial wall pyriform sinus cancer]. AB - OBJECTIVE: To study the surgical methods and functional outcome of preservative surgery for medial wall pyriform sinus cancer. METHODS: Seventy-one patients with medial wall pyriform sinus cancer, who were treated surgically between 1985 and 1997, were reviewed. Of the 71 cases, 49 underwent preservative surgery, and 22 total laryngectomy. Preservative surgical procedure was defined as follows: 1. Extent of resection: Supraglottic horizontal partial laryngectomy and resection of medial wall of pyriform sinus were performed in patients without fixation of the true vocal cord, and the section extended to paraglottic space, partial lateral wall of pyriform sinus, preepigottic space, superior-posterior of thyroid cartilage in those with restrained vocal cord motility. Supracricoid hemilaryngopharyngectomy and resection of medical wall and partial lateral wall of pyriform sinus were practiced in those with fixed hemilarynx. Cricoid ring and cervical esophagus were removed partially if the pyriform sinus apex was involved. 2. Reconstruction: Larynx: the remains of epiglottis, perichondrium of thyroid cartilage, the infrahyoid muscular fascia, and the platysmal flap were utilized to restore the defects of larynx. Pyriform sinus: Suturing the remaining pharyngeal mucosa directly to cover the wound if the defect was relatively small. For the large defect produced by extended resections, the pectoralis major myocutaneous flap and deltopectoral flap were used. 3. Surgical treatment of cervical lymphaden: Sixty five out of the 71 cases (91.5%) underwent neck dissection. Of which ipsilateral neck dissection were done in 39 cases, and bilateral neck dissection in 26 cases. 4. All patients received postoperative radiotherapy with doses of 60-75 Gy. RESULTS: In the group of preservative surgery the 3- and 5-year survival rates were 63.4% and 49.6% respectively, whereas those in the total laryngectomy group were 52.4% and 42.4% respectively. Of the 49 cases with preservative surgery, 71.4% (35/49) had all laryngeal functions restored and 28.6% (14/49) partially restored. CONCLUSION: Despite the fact that the medial wall pyriform sinus cancer tends to have laryngeal invasion, preservative surgery can be practiced for the majority of the cases with the lesions entirely removed. PMID- 12761990 TI - [Retrospective analysis of supracricoid partial laryngectomy with epiglottis preserved and reconstruction of laryngeal function]. AB - OBJECTIVE: To retrospectively evaluate the curative and functional consequences of supracricoid partial laryngectomy with epiglottis preserved. METHODS: From 1980 to 1996, 78 patients underwent supracricoid partial laryngectomy with the epiglottis preserved. The survival rate and the functional results were analyzed. RESULTS: Overall three-year survival rate was 87.2%(68/78), overall five-year survival rate was 75.6% (59/78), and ten-year survival rate was 21.1% (8/38). All patients resumed normal oral feeding. 97.4% (76/78) achieved tracheal extubation. Respiratory and speaking functions were good. CONCLUSION: Supracricoid partial laryngectomy with epiglottis preserved and reconstruction of laryngeal function not only improved local cancer control, but also preserves laryngeal physiologic function. PMID- 12761991 TI - [Effect of inhibition of polyamine biosynthesis on the growth and telomerase activity of human laryngeal carcinoma cells Hep-2]. AB - OBJECTIVE: To observe the effect of difluoromethylornithine (DFMO), an inhibitor of polyamine biosynthesis, on the growth and telomerase activity of human laryngeal carcinoma cells Hep-2, and try to find a new clue to search for the mechanism of reversion of tumor malignant phenotype via polyamine biosynthesis inhibition. METHODS: The growth characteristics of Hep-2 cells treated with DFMO were studied by morphological observation, cell growth curve and FCM analysis. Telomerase activity was detected by Telomerase Repeat Amplification Protocol (TRAP). RESULTS: The inhibition of cell proliferation, a marked increase in G1 and a decrease in S phase populations in the cell cycle of 2.5 mmol/L or 5 mmol/L DFMO treated Hep-2 cells were demonstrated; The cell apoptosis was shown by cell morphology and apoptosis peak in FCM; The telomerase activity of the treated cells was suppressed, while these changes were prevented by provision of exogenous putrescine(Pu) along with DFMO. CONCLUSION: Inhibition of polyamine biosynthesis could result in the proliferation inhibition and apoptosis induction of Hep-2 cells, which was associated with suppressed telomerase activity. It is suggested that inactivation of telomerase would be one of the important molecular events in reversion of tumor malignant phenotype via inhibition of polyamine biosynthesis. PMID- 12761992 TI - [Relationship among lymphatic metastasis, pericancerous lymphocytic reaction and dendritic cell infiltration in laryngeal carcinoma cells]. AB - OBJECTIVE: To study the role and the clinical significance of dendritic cell infiltration against tumor cells in patients with laryngeal squamous cell carcinoma. METHODS: Immunohistochemical method using S-100 protein antibody was employed to detect dendritic cells (DC) on paraffin-embedded tissue sections from 23 patients with laryngeal squamous cell carcinomas. The relationships among the density of dendritic cell infiltration in laryngeal carcinomas and cervical lymphatic metastasis, pericancerous lymphocytic reaction and other clinicopathologic parameters were observed. RESULTS: The numbers of dendritic cells infiltrating among laryngeal carcinoma cells in patients with no cervical lymph metastasis was significantly larger than those with cervical lymphatic metastasis (t = 4.889, P < 0.01). Significant increase DC infiltration among laryngeal carcinoma cells was found in the group with intensively positive reaction of pericancerous lymphocyte than in the medium and weakly positive groups. The number of infiltrating dendritic cells in patients who had survived more than 5 years was significantly larger than those survived less than 5 years (t = 4.423, P < 0.01). The numbers of the infiltrating dendritic cells in patients with well-differentiated squamous cell carcinoma and poorly differentiated squamous cells carcinoma were 2.2 +/- 1.07 and 14.6 +/- 7.14 respectively, which were significantly different (q = 4.532, P < 0.05). CONCLUSION: The study showed that DC infiltration among laryngeal squamous carcinoma cells played an important role in the host immune reaction against tumor. It indicated that when the density of infiltrating DC is higher, the patients had less chance of cervical lymph metastasis and may have a longer survival time. There was a coordinative effect between DC infiltration in tumor cells and pericancerous lymphocyte reaction. Thus, the DC infiltration among laryngeal squamous cells could be used as an index of prognosis. PMID- 12761993 TI - [Experimental study on polyaspartic acid inhibition of gentamicin-induced generation of reactive oxygen species in guinea pig cochlea]. AB - OBJECTIVE: To observe the polyaspartic acid(PAA) inhibition of gentamicin-induced reactive oxygen species generation in cochlea of guinea pig and to investigate the protective mechanism of polyaspartic acid on gentamicin ototoxicity. METHODS: Eighty-eight guinea pigs were divided randomly into four groups (GM, PAA + GM, PAA, and Saline). Gentamicin-induced reactive oxygen species (ROS) formation in cochlear tissue was detected directly with electron paramagnetic resonance (EPR) spectrometry at 1st, 5th and 10th day after administration of the drugs. At the same time, ABRs of guinea pigs were recorded and ultrastructural changes of lysosomes in the cochlear hair cells were observed with transmission electron microscopy. RESULTS: 1. At 1st day after administration of PAA and GM, there was some increase in EPR spectrometry in group GM and PAA + GM, There was no significant difference of ABR thresholds and ultrastructural changes of lysosomes in the cochlear hair cells among four groups(P > 0.05). 2. At 5th day, there was significant increase in of EPR spectrometry in group I (37.74 +/- 4.10, P < 0.01). At 10th day after administration of PAA and GM, there was no significant difference in EPR spectrometry among four groups (P > 0.05). In GM group, ultrastructural changes of lysosomes beneath cuticular plate of cochlear hair cells were more significant at 10th day than those at 5th day, including the increased number and volume of lysosome. In group GM, the longer the gentamicin administrated, the more significant increase in ABR thresholds had been noted. CONCLUSION: PAA significantly inhibits gentamicin-induced reactive oxygen species generation in cochlea of guinea pig, which showed that PAA has protective effect on gentamicin-ototoxicity and -phospholipidosis in guinea pigs. PMID- 12761994 TI - [Contralateral suppression of latency during distortion product otoacoustic emissions detection in guinea pigs]. AB - OBJECTIVE: To investigate the cochlear micromechanics and the function of cochlear efferent system by detecting the distortion product otoacoustic emissions (DPOAE) with or without contralateral acoustic stimulation. METHODS: The DPOAE amplitude and latency of the cubic DPOAE (2f1-f2) were measured at f2 = 2, 4 and 6 kHz with or without 70 dB SPL broad-band noise contralateral stimulation in ears of 12 healthy pigmented guinea pigs, which were divided into two groups(A and B) and tested with different primary intensity level combinations. On the next day, an operation was made to open the acoustic bulla of right ear in Group A. Kainic Acid (KA, 1 microL of 60 mmol/L) was applied onto the round window membrane and wiped up carefully after three hours. The amplitude and latency of the cubic DPOAE from the left ear were measured after 6 hours of KA administration, with or without contralateral noise stimulation. RESULTS: Before the KA treatment, the amplitude of DPOAE induced by primaries of equal levels (L1 = L2) presented no significant change and the latency prolonged significantly during contralateral acoustic stimulation (Group A). But the amplitude at 2, 4 kHz of DPOAE induced by primaries of different levels (L1 > L2) decreased significantly and the latency also prolonged significantly (Group B). After KA administration in the contralateral ear of Group A, the contralateral acoustic stimulation had no significant effect on either amplitude or latency of DPOAE measured ipsilaterally. CONCLUSION: The result demonstrates that the latency is a sensitive index for the cochlear contralateral suppression. The efferent system may act as a negative feedback during the cochlear transduction. PMID- 12761995 TI - [Measurement of nerve growth factor in the nasal mucosa of experimental allergic rhinitis]. AB - OBJECTIVE: To investigate the role of nerve growth factor in the development of allergic rhinitis. METHODS: Animal model of allergic rhinitis was established in rat, the content of nerve growth factor in the nasal mucosa was measured using enzyme-linked immunosorbent assay (ELISA) technique. RESULTS: The content of nerve growth factor significantly (P < 0.001) increased in the nasal mucosa of experimental allergic rhinitis (6.65 +/- 0.42) ng/g wet weight, compared with that in control animals (3.51 +/- 0.29) ng/g wet weight. CONCLUSION: Nerve growth factor may play an important role in the development of allergic rhinitis by regulating expression and release of neuropeptides and/or by activating immune cells such as mast cells. PMID- 12761996 TI - [Fabrication of allogeneic cartilage with tissue engineering technique and repair of laryngeal cartilage defects in rabbits]. AB - OBJECTIVE: To investigate the fabrication of allogeneic tissue engineered cartilage in predetermined shapes and its ability to repair laryngeal cartilage defects. METHODS: Tissue engineering technique was used to make allogeneic tissue engineered cartilage in predetermined shapes, and then some of them were utilized to repair cartilage defects in the rabbit larynges(n = 12). Finally, the chondrogenesis and reparative effect were studied grossly and histologically at various periods. RESULTS: 1. The gross specimens of 4 weeks after implantation revealed the presence of new hyaline cartilage of approximately the same shapes as original predetermined shapes, which had fair elasticity and support ability. Histological evaluation showed the characteristics of cartilage tissue. 2. Laryngeal cartilage defects were well repaired using allogeneic tissue engineered cartilage in different periods after operation. A few chondrocytes were observed between the reparative areas with tissue engineered cartilage and normal cartilage. Abundant matrices were also detected in the reparative and interfacial areas. No inflammatory cells were found. CONCLUSION: It is possible to regenerate new cartilage in predetermined shapes using tissue engineering technique in allograft animals with immunity. Allogeneic tissue engineered cartilage can be used as reparative material to repair cartilage defect of rabbit larynx. PMID- 12761997 TI - [Hydroxyapatite for experimental laryngotracheal reconstruction]. AB - OBJECTIVE: The porous hydroxyapatite (HA) tubiform prosthesis was employed for reconstruction of large circumferential defect of the larynx and cervical trachea. The biocompatibility, bioactivity and biofunctionality of the HA prosthesis were evaluated, and the feasibility for laryngotracheal reconstruction was discussed. METHODS: Twelve healthy canines were used to establish the experimental models, a large portion of cricoid cartilage and upper ten-ring segment of cervical trachea were resected and substituted with a corresponding 5 cm HA prosthesis by end-to-end anastomosis. Six months later, the eight survival canines were sacrificed and the HA prostheses with surrounding tissues were removed. They were observed in decalcified sections by optical microscopy and scanning electron microscopy. RESULTS: Two canines died in the immediate postoperative period with unknown reason and another two deaths were attributed to obstruction caused by the complete dislocation of the HA prostheses owing to rupture of the sutures within three weeks after operation. Eight canines survived up to six months. The implanted HA prostheses were tolerated in all cases without any rejection and dislocation or shift. An excellent airway was obtained and no signs of dyspnea and suffocation were found though there were hypergranulation and scar formation occurred at the site of anastomosis. Morphologic examination revealed that collagen fibers, new vessels and plenty of cells penetrated deeply into the pores of HA, and occupying the outer two third of HA wall. HA prostheses were surrounded by connective tissues and anchored firmly to the neighboring tissues, including the ends of the cricoid and tracheal cartilage by ingrowths of cartilaginous tissue into the macropores of the HA. However, the luminal surface of HA prosthesis was not covered at all section levels by respiratory mucosa. CONCLUSIONS: The implantation of the porous HA tubiform prosthesis can maintain the normal respiratory function of the experimental canines, but the proliferating granulation and scar formation at the anastomotic site are questions still remained to be solved. To cover the inner surface of HA with the epithelial mucosa and then reduce the morbidity caused by scar and hypergranulation, some forms of allografts such as pedicled flaps and jejunal autografts will be deserved. PMID- 12761998 TI - [Relationship between three inner ear antigens and autoimmune inner ear disease]. AB - OBJECTIVE: To investigate the relationship between three purified inner ear antigens and autoimmune inner ear disease (AIED), and to determine their expressions in normal guinea pig cochleas. METHODS: Guinea pigs were divided into group B, group C and group D and immunized respectively with three subcomponents of crude inner ear antigens (31,000, 42,000-45,000 and 60,000 proteins). Hearing thresholds, serum IgG levels and morphological changes of inner ear were observed. The protein expressions of these antigens were examined by immunohistochemistry. In the control group, antigens were replaced by gel homogenate of polyacrylamide. RESULTS: There were no significant differences of hearing threshold among groups before immunization with inner ear antigens (F = 0.07, P > 0.05). There were no significant changes of hearing thresholds and inner ear morphology in group C and control group. Hearing thresholds dropped in part of animals immunized with 31,000 or 60,000 protein. Hearing threshold of group B and D changed significantly after immunization, as compared with the control group(F = 9.12, P < 0.01). Serum IgG levels increased significantly in all experimental groups compared with the control group(F = 7.46, P < 0.01). The 31,000 protein distributed strictly in cochlear nerve, and 42,000-45,000 or 60,000 protein distributed widely, including the spiral ganglion, Corti's organ, stria vascularis and spiral ligament. CONCLUSIONS: Two subcomponents of 31,000 and 60,000 in crude inner ear antigen could induce autoimmune inner ear disease. The distribution of 31,000 protein was more tissue specific and might be used as a marker protein for clinical diagnosis of autoimmune inner ear disease. PMID- 12761999 TI - [A retrospective study on cholesteatoma otitis media coexisted with cholesterol granuloma]. AB - OBJECTIVE: To investigate the etiology and pathogenesis of cholesteatoma otitis media accompanied by cholesterol granuloma and the relationship between cholesteatoma and cholesterol granuloma. METHODS: Sixty three cases of middle ear cholesterol granuloma treated in our hospital during the period from March 1988 to May 2000 were retrospectively reviewed. All cases were verified by surgery and pathology. Fifteen cases of cholesteatoma coexisted with cholesterol granuloma were found among the 63 patients. RESULTS: All fifteen cases had a long-term history of otitis media, such as otorrhea (sanguine purulent otorrhea and bloody otorrhea in 8 cases) and perforation of the ear drum (perforation of pars flaccida in 8 cases). Temporal bone CT scans showed cholesteatoma in 11 cases. All patients were treated surgically, and cholesteatoma and cholesterol granuloma were found coexisting alternatively, and the latter lied mainly in the tympanic antrum, attic and mastoid air cells. Chocolate-colored mucus was accumulated in well-developed mastoid air cells, and glistening dotty cholesterol crystals were also found. In most cases, enlarged aditus, destruction of lateral attic wall, erosion of ossicular chain, exposition of horizontal segment of facial nerve and tegmen of attic were found. Occlusion of Eustachian tube was found in 6 cases, and occlusion of tympanic isthmus was found in all cases. A post-operative dry ear was obtained, and hearing improved in all 12 cases following tympanoplasty. CONCLUSIONS: Cholesteatoma and cholesteatoma granuloma in middle ear may share a common pathophysiological etiology, an occlusion of ventilation and a disturbance of drainage. The diagnosis should be considered when patients with chronic otitis media presented with bloody otorrhea. CT and magnetic resonance imaging(MRI) are useful for the diagnosis before operation. The surgical approach depends on the location, extension and severity of the lesion, and the principle of surgery is to clear the lesion and create an adequate drainage. PMID- 12762000 TI - [A preliminary study of a hearing screening model for newborn]. AB - OBJECTIVE: To search for a hearing screening model for newborn and to investigate the prevalence of newborn hearing loss in our country. METHODS: The distortion product otoacoustic emissions (DPOAE) was used to test the hearing in 2,998 of 3,075 newborns before discharge. Otoacoustic emissions (OAE) was again used for cases failed in the hospital screening 4 weeks later. Those cases failed in both screening steps were finally tested by auditory brainstem response (ABR). All infants failed in ABR test received diagnostic evaluation audiologically to identify the category and degree of hearing loss. The pass criterion of DPOAE was defined as signal-noise-ratio (SNR) exceeding 6 dB in 4 of 5 frequencies between 1.5-6 kHz. The pass criterion of ABR was the presence of wave V in response to 35 dB nHL click stimuli. RESULTS: The OAE screening in the hospital showed that 2,710 (90.4%) newborns passed the first test. Two hundred and sixty three of 288 newborns passed the second OAE screening after one month. Six of 25 infants failed in ABR test were eventually identified to be hearing impaired. CONCLUSION: Two-stages screening, combining OAE and ABR tests, may be an ideal model for newborn hearing screening. The prevalence of congenital hearing loss is similar to that reported in the literature. PMID- 12762001 TI - [Clinical observation on sensorineural hearing loss secondary to secretory otitis media]. AB - OBJECTIVE: To study whether secretory otitis media causes sensorineural deafness or not, especially in stubborn cases. METHODS: The 0.5 kHz, 1 kHz, 2 kHz, 4 kHz BC stimuli were administered to 164 patients with secretory otitis media, and their BC loss calculated. As a control, BC threshold was also recorded in the contralateral ear of 66 unilaterally deaf patients. RESULTS: BC threshold was raised in 94 out of 164 patients (57.3%). There was no statistically significant difference between bilaterally and unilaterally deaf groups. All patients had similar BC hearing loss in the same frequency, but had different BC hearing loss in different frequencies and the most serious loss was in 4 kHz. CONCLUSION: The rate of sensorineural deafness caused by secretory otitis media is higher than expected. Frequency and intensity of hearing loss between unilaterally and bilaterally deaf patients with secretory otitis media were no different. All ears had different hearing loss in different frequencies, but the most serious hearing loss was in high frequency and it had the tendency to spread to the speech frequency. PMID- 12762002 TI - [Expression of the inducible isoform of nitric oxide synthase mRNA and the role in nasal polyps]. AB - OBJECTIVE: To explore the pathogenesis of nasal polyps. The aim of this study is to detect the expression of inducible isoform of nitric oxide synthase (iNOS) in nasal polyp tissue, and to compare these findings with that in normal nasal turbinate. METHODS: We examined the expression of iNOS in human nasal polyps from patients undergoing nasal polypectomy (n = 8) and nasal turbinectomy(n = 6). The iNOS mRNA expression was determined by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). RESULTS: The iNOS mRNA expression was significantly stronger in the nasal polyps (NP) than that in turbinate tissues (P < 0.01). CONCLUSIONS: iNOS expression is upregulated in NP, which indicate that iNOS and NO may play a potential role in the formation and growth of NP. PMID- 12762003 TI - [Determining the site of airway obstruction in obstructive sleep apnea with airway pressure measurements during sleep]. AB - OBJECTIVE: Determine the site(s) of upper airway obstruction and the influence of sleep stage on the pattern of obstruction with upper airway (UA) pressure measurement. Analyses the clinical value of UA pressure measurement and it's influenced factors. METHODS: 30 OSAS patients underwent UA pressure measurement during polysomnography. Multisensory pressure catheters with five solid-state ultraminiature sensors were insert through patients' upper airway to esophagus, The sensors were located at the nasopharynx, oropharynx, tongue base, hypopharynx and esophagus and the lower limit of UA obstruction was determined relying on the observed pressure pattern. RESULTS: (1) During inspiration, obstruction occurred associated with a increased negative inspiratory pressure inferior to the site of obstruction and a disappeared negative inspiratory pressure above the site of obstruction. (2) Three patterns of obstruction were observed. 1. The site of obstruction was located at the level of the palate. 2. The level of the palate and tonguebase obstruction all present. 3. Nasopharyngeal obstruction soft plate obstruction and tongue-pharynx obstruction all present. (3) The factors which affected these measurement are 1. The catheter were placed by observation with a fiberoptic endoscope to keep the sensors located at the correct position. 2. Catheter plugging with secretions need to be prevented. CONCLUSION: UA pressure measurement can objectively identify the level of obstruction during sleep. PMID- 12762004 TI - RNA structure and the roots of protein synthesis. PMID- 12762005 TI - Atomic structures of the 30S subunit and its complexes with ligands and antibiotics. PMID- 12762006 TI - Progress toward an understanding of the structure and enzymatic mechanism of the large ribosomal subunit. PMID- 12762007 TI - High-resolution structures of ribosomal subunits: initiation, inhibition, and conformational variability. PMID- 12762008 TI - Structure of the ribosome at 5.5 A resolution and its interactions with functional ligands. PMID- 12762009 TI - Ratchet-like movements between the two ribosomal subunits: their implications in elongation factor recognition and tRNA translocation. PMID- 12762010 TI - Do single (ribosome) molecules phase themselves? PMID- 12762011 TI - Correlating the X-ray structures for halo- and thermophilic ribosomal subunits with biochemical data for the Escherichia coli ribosome. PMID- 12762012 TI - Probing ribosome structure and function by mutagenesis. PMID- 12762013 TI - Exploring the mechanism of the peptidyl transfer reaction by chemical footprinting. PMID- 12762014 TI - Analysis of the active site of the ribosome by site-directed mutagenesis. PMID- 12762015 TI - Codon recognition and decoding: the transorientation hypothesis. PMID- 12762016 TI - Features and functions of the ribosomal E site. PMID- 12762017 TI - Pseudouridines and pseudouridine synthases of the ribosome. AB - psi are ubiquitous in ribosomal RNA. Eubacteria, Archaea, and eukaryotes all contain psi, although their number varies widely, with eukaryotes having the most. The small ribosomal subunit can apparently do without psi in some organisms, even though others have as many as 40 or more. Large subunits appear to need at least one psi but can have up to 50-60. psi is made by a set of site specific enzymes in eubacteria, and in eukaryotes by a single enzyme complexed with auxiliary proteins and specificity-conferring guide RNAs. The mechanism is not known in Archaea, but based on an analysis of the kinds of psi synthases found in sequenced archaeal genomes, it is likely to involve use of guide RNAs. All psi synthases can be classified into one of four related groups, virtually all of which have a conserved aspartate residue in a conserved sequence motif. The aspartate is essential for psi formation in all twelve synthases examined so far. When the need for psi in E. coli was examined, the only synthase whose absence caused a major decrease in growth rate under normal conditions was RluD, the synthase that makes psi 1911, psi 1915, and psi 1917 in the helix 69 end loop. This growth defect was the result of a major failure in assembly of the large ribosomal subunit. The defect could be prevented by supplying the rluD structural gene in trans, and also by providing a point mutant gene that made a synthase unable to make psi. Therefore, the RluD synthase protein appears to be directly involved in 50S subunit assembly, possibly as an RNA chaperone, and this activity is independent of its ability to form psi. This result is not without precedent. Depletion of PET56, a 2'-O-methyltransferase specific for G2251 (E. coli numbering) in yeast mitochondria virtually blocks 50S subunit assembly and mitochondrial function (Sirum-Connolly et al. 1995), but the methylation activity of the enzyme is not required (T. Mason, pers. comm.). The absence of FtsJ, a heat shock protein that makes Um2552 in E. coli, makes the 50S subunit less stable at 1 mM Mg++ (Bugl et al. 2000) and inhibits subunit joining (Caldas et al. 2000), but, in this case, it is not yet known whether the effects are due to the lack of 2'-O-methylation or to the absence of the enzyme itself. Is there any role for the psi residues themselves? First, as noted above, the 3 psi made by RluD which cluster in the end-loop of helix 69 are highly conserved, with one being universal (Fig. 2B). In the 70S-tRNA structure (Yusupov et al. 2001), the loop of this helix containing the psi supports the anticodon arm of A-site tRNA near its juncture with the amino acid arm. The middle of helix 69 does the same thing for P-site tRNA. Unfortunately, the resolution is not yet sufficient to provide a more precise alignment of the psi residues with the other structural elements of the tRNA-ribosome complex so that one cannot yet determine what role, if any, is played by the N-1 H that distinguishes psi from U. Second, and more generally, some psi residues in the LSU appear to be near the site of peptide bond formation or tRNA binding but not actually at it (Fig. 2B) (Nissen et al. 2000; Yusupov et al. 2001). For example, position 2492 is commonly psi and is only six residues away from A2486, the A postulated to catalyze peptide-bond formation. Position 2589 is psi in all the eukaryotes and is next to 2588, which base-pairs with the C75 of A-site tRNA. Residue 2620, which interacts with the A76 of A-site-bound tRNA, is a psi or is next to a psi in eukaryotes and Archaea, and is five residues away from psi 2580 in E. coli. A2637, which is between the two CCA ends of P- and A-site tRNA, is near psi 2639, psi 2640, and psi 2641, found in a number of organisms. Residue 2529, which contacts the backbone of A site tRNA residues 74-76, is near psi 2527 psi 2528 in H. marismortui. Residues 2505-2507, which contact A-site tRNA residues 50-53, are near psi 2509 in higher eukaryotes, and residues 2517-2519 in contact with A-site tRNA residues 64-65 are within 1-3 nucleotides of psi 2520 in higher eukaryotes and psi 2514 in H. marismortui. A way to rationalize this might be to invoke the concept suggested in the Introduction that psi acts as a molecular glue to hold loose elements in a more rigid configuration. It may well be that this is more important near the site of peptide-bond formation and tRNA binding, accounting for the preponderance of psi in this vicinity. What might be the role of all the other psi in eukaryotes? One can only surmise that cells, having once acquired the ability to make psi with guide RNAs, took advantage of the system to inexpensively place psi wherever an undesirable loose region was found. It might be that in some of these cases, psi performs the role played by proteins in other regions, namely that of holding the rRNA in its proper configuration. Confirmation of this hypothesis will have to await structural determination of eukaryotic ribosomes. PMID- 12762018 TI - Formation of two classes of tRNA synthetases in relation to editing functions and genetic code. PMID- 12762019 TI - Structural basis for amino acid and tRNA recognition by class I aminoacyl-tRNA synthetases. PMID- 12762020 TI - Aminoacyl-tRNA synthesis: a postgenomic perspective. PMID- 12762021 TI - tRNA conformity. PMID- 12762022 TI - Initiator tRNA and its role in initiation of protein synthesis. PMID- 12762023 TI - On translation by RNAs alone. PMID- 12762024 TI - Overriding standard decoding: implications of recoding for ribosome function and enrichment of gene expression. PMID- 12762025 TI - Structure and function of the stimulatory RNAs involved in programmed eukaryotic 1 ribosomal frameshifting. PMID- 12762026 TI - Programmed +1 translational frameshifting in the yeast Saccharomyces cerevisiae results from disruption of translational error correction. PMID- 12762027 TI - Preferential translation of adenovirus mRNAs in infected cells. PMID- 12762028 TI - Shunting and controlled reinitiation: the encounter of cauliflower mosaic virus with the translational machinery. PMID- 12762029 TI - Mechanisms of internal ribosome entry in translation initiation. PMID- 12762030 TI - Initiator Met-tRNA-independent translation mediated by an internal ribosome entry site element in cricket paralysis virus-like insect viruses. PMID- 12762031 TI - The mRNA closed-loop model: the function of PABP and PABP-interacting proteins in mRNA translation. PMID- 12762032 TI - The mRNA capping apparatus as drug target and guide to eukaryotic phylogeny. PMID- 12762033 TI - Nonsense-mediated mRNA decay: insights into mechanism from the cellular abundance of human Upf1, Upf2, Upf3, and Upf3X proteins. PMID- 12762034 TI - Nonsense-mediated mRNA decay in Saccharomyces cerevisiae: a quality control mechanism that degrades transcripts harboring premature termination codons. PMID- 12762035 TI - Translational control of 15-lipoxygenase and msl-2 mRNAs: single regulators or corepressor assemblies? PMID- 12762036 TI - PUF proteins and 3'UTR regulation in the Caenorhabditis elegans germ line. PMID- 12762037 TI - Translational control of embryonic cell division by CPEB and maskin. PMID- 12762038 TI - Small RNA regulators of translation: mechanisms of action and approaches for identifying new small RNAs. PMID- 12762039 TI - Initiation factors in the early events of mRNA translation in bacteria. PMID- 12762040 TI - The translation of capped mRNAs has an absolute requirement for the central domain of eIF4G but not for the cap-binding initiation factor eIF4E. PMID- 12762041 TI - Functions of eukaryotic factors in initiation of translation. PMID- 12762042 TI - Interactions of the eukaryotic translation initiation factor eIF4E. PMID- 12762043 TI - A multifactor complex of eIF1, eIF2, eIF3, eIF5, and tRNA(i)Met promotes initiation complex assembly and couples GTP hydrolysis to AUG recognition. PMID- 12762044 TI - Universal translation initiation factor IF2/eIF5B. PMID- 12762045 TI - Structural studies of eukaryotic elongation factors. PMID- 12762047 TI - Mechanism of elongation factor G function in tRNA translocation on the ribosome. PMID- 12762046 TI - Translation elongation factor 1 functions in the yeast Saccharomyces cerevisiae. PMID- 12762048 TI - Modulation of translation termination mechanisms by cis- and trans-acting factors. PMID- 12762049 TI - Protein tRNA mimicry in translation termination. PMID- 12762050 TI - Mitogenic and nutritional signals are transduced into translational efficiency of TOP mRNAs. PMID- 12762051 TI - Double-stranded RNA-binding proteins and the control of protein synthesis and cell growth. PMID- 12762052 TI - Translational regulation in the cellular response to biosynthetic load on the endoplasmic reticulum. PMID- 12762053 TI - A novel stress-response protein that binds at the ribosomal subunit interface and arrests translation. PMID- 12762054 TI - The fourth step of protein synthesis: disassembly of the posttermination complex is catalyzed by elongation factor G and ribosome recycling factor, a near-perfect mimic of tRNA. PMID- 12762055 TI - Structure, function, and regulation of free and membrane-bound ribosomes: the view from their substrates and products. PMID- 12762056 TI - The active 80S ribosome-Sec61 complex. PMID- 12762057 TI - Ribosomal RNA genes, RNA polymerases, nucleolar structures, and synthesis of rRNA in the yeast Saccharomyces cerevisiae. PMID- 12762058 TI - Economics of ribosome biosynthesis. PMID- 12762059 TI - Ribosome biogenesis: role of small nucleolar RNA in maturation of eukaryotic rRNA. PMID- 12762060 TI - Thermodynamics and kinetics of central domain assembly. PMID- 12762061 TI - Nuclear export of the large ribosomal subunit. PMID- 12762062 TI - The ribosome in the 21st century: the post-structural era. PMID- 12762063 TI - [Diagnostic value of 99mTc-methoxy-isobutyl-isonitril and 99mTc-methylene diphosphonate scintimammography in the preoperative evaluation of breast cancer]. AB - AIM: To compare and establish the value of 99mTc-MIBI (methoxyisobutiylisonitrile) and 99mTc-MDP (methylene diphosphonate) scintigraphy in patients with suspicion of breast cancer. PATIENTS AND METHODS: The study included 105 patients. All patients were evaluated before surgery by 99mTc-MIBI and 99mTc-MDP scintigraphy. Thoracic SPECT study (Multispect II, Siemens) was performed 10 min. after i.v. injection of 99mTc-MIBI and 2 hours after i.v. injection of 99mTc-MDP. Final diagnosis was achieved after operation by histology. RESULTS: 99mTc-MIBI scintigraphy showed a better sensitivity (87%) than the 99mTc-MDP scintigraphy (81%) and the results are similar in specificity and validity, too. CONCLUSIONS: The results suggest that 99mTc-MIBI scintigraphy is a useful method in the preoperative diagnosis of patients with suspicion of breast cancer. PMID- 12762064 TI - [Recent developments in the diagnosis of cutaneous manifestations of internal diseases]. AB - Remarkable advances in the field of diagnostics have been made over the last decades. Therefore, more attention is focused on the changes in the relationship between the skin symptoms (clues) and the pathology of the internal organs. The present paper highlights the following trends: 1. Changes in the concept of the correlation dermatologic diseases. 2. Coincidence of the symptoms typical of a disease in other diseases affecting the internal organs. 3. Recognition of new relationships of the correlation dermatology. 4. A scope of new indicator symptoms. 5. Occurrence of the "sine morbo" dermatodromas. PMID- 12762065 TI - [Postoperative renal failure and its risk factors after open heart surgery]. AB - AIM: In this retrospective study the authors have investigated the occurrence and the possible risk factors of renal failure following heart surgery on CPB. PATIENTS AND METHODS: 52 perioperative variables of 536 consecutive patients undergoing cardiac surgery have been analysed. Types of surgery were as follows: coronary-bypass: 266 (49.6%), combined coronary-bypass: 62 (11.5%), valve replacement: 171 (31.9%), adult repair of congenital: disease 24 (4.4%), aortic dissection: 6 (1.1%), others 7 (1.3%). In the past medical history those patients involved in the study chronically impaired renal function could be found in 2.2%, whilst kidney stone and chronic pyelonephritis appeared in 2.4% and 1.2% respectively. Postoperative renal failure developed in 31 patients (5.8%), 3 of them required haemodialysis (0.6%). For the statistical analysis chi 2 test, t test, Levine-test, Mann-Whitney-test and logistic regression analysis were applied using the SPSS software. RESULTS: On the basis of the performed multivariable logistic regression analysis the risk factors of renal failure following open heart surgery are as follows: age, duration of anaesthesia, chronic pyelonephritis in past medical history, preoperative serum creatinine level and low cardiac output syndrome. PMID- 12762066 TI - [Mini-Cog: a simple method for very brief screening of mental decline]. AB - INTRODUCTION: Physicians do not detect majority of the demented people, and many patients appear only in an advanced stage of their mental decline at the first medical visit. Role of general practitioner is crucial; a brief and effective screening method can basically change the present situation. Borson et al (2000) has investigated a new, very brief method named Mini-Cog that consists of the combination of the 3-word recall and the clock-drawing test. AIM: of the present study was to evaluate this method in Hungarian patient population. METHODS: Authors have analysed the charts of patients examined by the first author in the outpatient part of the Memory clinic of the National Institute of Psychiatry and Neurology in a 2-year interval (between 11.01. 1999 and 31.10.2001). Data of 186 patients were analysed. From the Mini-Mental State the total score, score of three-word recall (possible scores between 0-3), serial counting backward with 7 from 100 were taken into consideration; the clock-drawing test was analysed quantitatively (1 = failure, 2 = borderland, 3 = acceptable solution). RESULTS: The algorithm which considers the patient demented if she or he does not remember of any word, or remembers only of 1-2 and the clock-drawing is pathologic, has a high sensitivity (100%) and specificity (80.9%) in cases using the < or = 24 Mini Mental State value, while the sensitivity and specificity is 98.8% and 88.0% respectively if drawing the limit at < or = 26 points. Combination of the two tests (word-recall, clock drawing) gives a better value than either of them alone. Based on their experiences they propose to apply in a short test the serial backwards with seven from 100 too, while patients with very high Mini Mental State scores (29-30) not infrequently can not perfectly solve this subtest (7/11 = 64 p.c.). CONCLUSION: In agreement with the Hungarian Protocol for diagnosing and treating dementias (1999) authors stress the importance of the consultation with a specialist (neurologist or psychiatrist) in cases of suspected dementia to reveal the underlying disease. PMID- 12762067 TI - [Acute porphyrias in differential diagnosis]. AB - The characteristic symptoms for acute porphyrias are caused by the inherited decreased activity of the enzymes of the heme biosynthesis pathway. Usually there is an exogenous or endogenous factor inhibiting the heme biosynthesis or increasing the consumption of heme produced in already decreased amount. The most important precipitating factors are the therapeutic drugs. Therefore, certain therapeutic drugs ordered for carriers or patients with acute porphyria are serious risk factors. It is very important to identify patients and carriers with acute porphyria as early as possible and to make a close follow-up so the development of the symptoms of the life threatening acute attack could be prevented. It is very difficult to suspect the diagnosis of acute porphyria. There is a very characteristic discrepancy between the serious complaints and the actual clinical findings. The severe cramping abdominal pain, nausea, vomiting, muscle weakness of the limbs and sensory loss are the main signs at the beginning. The specific symptoms which help to establish the diagnosis--red colored urine, hyponatremia, tachycardia, hypertension, subileus, acute psychosis, gradually developing paresis of the lower and then the upper limbs- are characteristic for the later phase of the acute attack. Very often there is a rapid progression with Landry-type paralysis developing in days or even in hours, following respiratory paralysis or serious arrhythmia is the cause of the death. In case of suspicion of acute porphyria the patient should be directed to a department where the specific laboratory methods--measurement of the porphyrin precursors, porphyrins and their isomers in urine and feces, quantitation of protoporphyrin in red blood cells, measurement of the plasma porphyrin and enzyme activity--to diagnose the different types of the disease and the immediate specific treatment with heme arginate are possible if needed. All of these are available in the National Porphyria Center. PMID- 12762068 TI - [Revolution and tragedy in the history of obstetrics: the life and work of Semmelweis--with today's eyes]. PMID- 12762069 TI - Health effects of local residents near the World Trade Center: have they been forgotten? PMID- 12762070 TI - Sensory irritation testing. PMID- 12762071 TI - The role of mercury in pustulosis palmaris et plantaris. PMID- 12762072 TI - Adverse human health effects associated with molds in the indoor environment. AB - Molds are common and important allergens. About 5% of individuals are predicted to have some allergic airway symptoms from molds over their lifetime. However, it should be remembered that molds are not dominant allergens and that the outdoor molds, rather than indoor ones, are the most important. For almost all allergic individuals, the reactions will be limited to rhinitis or asthma; sinusitis may occur secondarily due to obstruction. Rarely do sensitized individuals develop uncommon conditions such as ABPA or AFS. To reduce the risk of developing or exacerbating allergies, mold should not be allowed to grow unchecked indoors. When mold colonization is discovered in the home, school, or office, it should be remediated after the source of the moisture that supports its growth is identified and eliminated. Authoritative guidelines for mold remediation are available. Fungi are rarely significant pathogens for humans. Superficial fungal infections of the skin and nails are relatively common in normal individuals, but those infections are readily treated and generally resolve without complication. Fungal infections of deeper tissues are rare and in general are limited to persons with severely impaired immune systems. The leading pathogenic fungi for persons with nonimpaired immune function, Blastomyces, Coccidioides, Cryptococcus, and Histoplasma, may find their way indoors with outdoor air but normally do not grow or propagate indoors. Due to the ubiquity of fungi in the environment, it is not possible to prevent immunecompromised individuals from being exposed to molds and fungi outside the confines of hospital isolation units. Some molds that propagate indoors may under some conditions produce mycotoxins that can adversely affect living cells and organisms by a variety of mechanisms. Adverse effects of molds and mycotoxins have been recognized for centuries following ingestion of contaminated foods. Occupational diseases are also recognized in association with inhalation exposure to fungi, bacteria, and other organic matter, usually in industrial or agricultural settings. Molds growing indoors are believed by some to cause building-related symptoms. Despite a voluminous literature on the subject, the causal association remains weak and unproven, particularly with respect to causation by mycotoxins. One mold in particular, Stachybotrys chartarum, is blamed for a diverse array of maladies when it is found indoors. Despite its well-known ability to produce mycotoxins under appropriate growth conditions, years of intensive study have failed to establish exposure to S. chartarum in home, school, or office environments as a cause of adverse human health effects. Levels of exposure in the indoor environment, dose-response data in animals, and dose-rate considerations suggest that delivery by the inhalation route of a toxic dose of mycotoxins in the indoor environment is highly unlikely at best, even for the hypothetically most vulnerable subpopulations. Mold spores are present in all indoor environments and cannot be eliminated from them. Normal building materials and furnishings provide ample nutrition for many species of molds, but they can grow and amplify indoors only when there is an adequate supply of moisture. Where mold grows indoors there is an inappropriate source of water that must be corrected before remediation of the mold colonization can succeed. Mold growth in the home, school, or office environment should not be tolerated because mold physically destroys the building materials on which it grows, mold growth is unsightly and may produce offensive odors, and mold is likely to sensitize and produce allergic responses in allergic individuals. Except for persons with severely impaired immune systems, indoor mold is not a source of fungal infections. Current scientific evidence does not support the proposition that human health has been adversely affected by inhaled mycotoxins in home, school, or office environments. PMID- 12762073 TI - Effects of the GSTM1 and GSTT1 polymorphisms on the relationship between maternal exposure to environmental tobacco smoke and neonatal birth weight. AB - The purpose of the investigation was to determine whether genetic polymorphisms in enzymes that metabolize exogenous chemicals modulate the effects of environmental tobacco smoke (ETS) exposure on birth weight. A survey was conducted from 2000 to 2001 among 266 pregnant women who were hospitalized for delivery and on their singleton live births. We determined maternal GSTM1 and GSTT1 polymorphisms by polymerase chain reaction and measured the urinary cotinine of pregnant women at delivery by radioimmunoassay. Birth weight was found to decrease significantly with increasing concentrations of maternal urinary cotinine (P < 0.05). The interactive effect of exposure to ETS and the presence of the GSTT1 polymorphism was found to be significant by multivariate analysis (P < 0.01), whereas the interactive effect of exposure to ETS and the presence of GSTM1 polymorphism did not reach statistical significance (P = 0.21). A combination of the GSTM1-null and the GSTT1 null-genotypes was found to exacerbate the effect of maternal exposure to ETS on birth weight more than the presence of either genotype alone. Our data indicate that maternal exposure to ETS negatively affects neonatal birth weight, and the adverse effect of maternal exposure to ETS on neonatal birth weight could be modified by the maternal metabolic genotypes, GSTM1 and GSTT1. PMID- 12762074 TI - Early workplace intervention for employees with musculoskeletal-related absenteeism: a prospective controlled intervention study. AB - Sickness absenteeism caused by musculoskeletal disorders (MSDs) is a persistent and costly occupational health challenge. In a prospective controlled trial, we compared the effects on sickness absenteeism of a more proactive role for insurance case managers as well as workplace ergonomic interventions with that of traditional case management. Patients with physician-diagnosed MSDs were randomized either to the intervention group or the reference group offered the traditional case management routines. Participants filled out a comprehensive questionnaire at the initiation of the study and after 6 months. In addition, administrative data were collected at 0.6, and 12 months after the initiation of the project. For the entire 12-month period, the total mean number of sick days for the intervention group was 144.9 (SEM 11.8) days/person as compared to 197.9 (14.0) days in the reference group (P < 0.01). Compared with the reference group, employees in the intervention group significantly more often received a complete rehabilitation investigation (84% versus 27%). The time for doing this was reduced by half (59.4 (5.2) days versus 126.8 (19.2), P < .01). The odds ratio for returning to work in the intervention group was 2.5 (95% confidence interval 1.2-5.1) as compared with the reference group. The direct cost savings were USD 1195 per case, yielding a direct benefit-to-cost ratio of 6.8. It is suggested that the management of MSDs should to a greater degree focus on early return to work and building on functional capacity and employee ability. Allowing the case managers a more active role as well as involving an ergonomist in workplace adaptation meetings might also be beneficial. PMID- 12762075 TI - Ten years' experience using an integrated workers' compensation management system to control workers' compensation costs. AB - This work presents 10 years of experience using an Integrated Workers' Compensation Claims Management System that allows safety professionals, adjusters, and selected medical and nursing providers to collaborate in a process of preventing accidents and expeditiously assessing, treating, and returning individuals to productive work. The hallmarks of the program involve patient advocacy and customer service, steerage of injured employees to a small network of physicians, close follow-up, and the continuous dialogue between parties regarding claims management. The integrated claims management system was instituted in fiscal year 1992 servicing a population of approximately 21,000 individuals. The system was periodically refined and by the 2002 fiscal year, 39,000 individuals were managed under this paradigm. The frequency of lost-time and medical claims rate decreased 73% (from 22 per 1000 employees to 6) and 61% (from 155 per 1000 employees to 61), respectively, between fiscal year 1992 and fiscal year 2002. The number of temporary/total days paid per 100 insureds decreased from 163 in fiscal year 1992 to 37 in fiscal year 2002, or 77%. Total workers' compensation expenses including all medical, indemnity and administrative, decreased from $0.81 per $100 of payroll in fiscal year 1992 to $0.37 per $100 of payroll in fiscal year 2002, a 54% decrease. More specifically, medical costs per $100 of payroll decreased 44% (from $0.27 to $0.15), temporary/total, 61% (from $0.18 to $0.07), permanent/partial, 63% (from $0.19 to $0.07) and administrative costs, 48% ($0.16 to $0.09). These data suggests that workers' compensation costs can be reduced over a multi-year period by using a small network of clinically skilled health care providers who address an individual workers' psychological, as well as physical needs and where communication between all parties (e.g., medical care providers, supervisors, and injured employees) is constantly maintained. Furthermore, these results can be obtained in an environment in which the employer pays the full cost of medical care and the claimant has free choice of medical provider at all times. PMID- 12762076 TI - Workplace accommodations for people with disabilities: National Health Interview Survey Disability Supplement, 1994-1995. AB - As American workers age, workers with impairments and functional limitations make up a larger percentage of our workforce. This investigation presents data from the National Health Interview Survey Disability Supplement 1994-1995 (NHIS-D) describing the nature of workplace accommodations in the American workforce and factors associated with the provision of such accommodations. Of a nationally representative sample of workers aged 18 to 69 years with a wide range of impairments, 12% reported receiving workplace accommodations. Males (odds ratio (OR) 0.64: 95% confidence interval (CI) 0.53-0.78) and Southerners (OR 0.57; 95% CI = 0.47-0.70) were less likely than others to receive workplace accommodations. Those with mental health conditions were less likely than others to receive accommodations (OR 0.56; 95% CI = 0.44-0.70). College graduates (OR 1.53; 95% CI = 1.22-1.91), older workers, full time workers (OR 3.99; 95% CI = 2.63-3.87), and the self-employed (OR 1.76; 95% CI = 1.28-2.41) were more likely than others to receive accommodations. PMID- 12762077 TI - Possible effects of polychlorinated biphenyls and organochlorinated pesticides on the thyroid after long-term exposure to heavy environmental pollution. AB - The purpose of this work was to study the effects of high environmental exposure to polychlorinated biphenyls (PCBs) and other organochlorines on the thyroid. Thyroid volume, hypoechogenicity and nodules (by ultrasound), presence of antithyroid peroxidase (anti-TPO) antibodies, and abnormal thyroid-stimulating hormone (TSH) levels in serum (by radioimmunoassay) were examined in 101 adults from the PCB-polluted area in 360 controls. Serum levels of PCBs, hexachlorobenzene, gamma-hexachlorocyclohexane (HCH), p,p'-DDT(1,1,1-trichloro 2,2'-bis(p-chlorophenyl)ethane), and p,p'-DDE(1,1-dichloro-2,2'-bis(p chlorophenyl)ethene) were measured by high-resolution gas chromatography. Very high levels of PCBs were found in the polluted area (7300 +/- 871 ng/g lipids) compared with controls (2045 +/- 147 ng/g). Positive correlations (P < 0.001) were found between the levels of all organochlorines and their total except for hexachlorocyclohexane (HCH). In the polluted area, the highest thyroid volumes (18.7 +/- 2.32 mL; mean +/- SE) were clustered among 23 subjects (17 males and six females) with PCB levels above 10,000 ng/g (range 10,000-58,667 ng/g). In the remaining 438 subjects the thyroid volume was 14.2 +/- 0.29 mL. These data suggest that there might be a threshold serum PCB level of approximately 10,000 ng/g that may influence the thyroid volume. A two-way ANOVA showed that all thyroid volumes in the polluted area were significantly higher (P < 0.001) than in the control area. In males from the polluted area, the frequencies of thyroid hypoechogenicity, thyroid nodules, positive anti-TPO, and abnormal TSH level were higher than in males from the control area, whereas such differences were not observed in females. Increased thyroid volume and indicators of potential thyroid dysfunction were associated with long-term environmental exposure to PCBs. These effects on the thyroid were confined to subjects with PCB levels above 10,000 ng/g of lipid (thyroid volume) and to males from the polluted area (thyroid hypoechogenicity, thyroid nodules, positive anti-TPO, and abnormal TSH). PMID- 12762078 TI - Vinyl chloride and liver and brain cancer at a polymer production plant in Louisville, Kentucky. AB - Studies of overlapping cohorts in the United States have shown an excess mortality from brain cancer in vinyl chloride (VC)-exposed workers. One plant located in Louisville, Kentucky, is included in many of these studies. We separated this plant from the large US cohort and re-analyzed the mortality experience. Louisville experienced significantly elevated liver (standardized mortality analyses [SMR] = 400) and brain cancer (SMR = 229) mortality. Liver cancer mortality remained significantly elevated (SMR = 344) in the remaining cohort; however, brain cancer mortality was markedly reduced (SMR = 112) when Louisville was removed. In contrast with liver cancer, a preliminary review of work assignments did not suggest that the brain cancer excess was related to VC exposure. The Louisville brain cancer cluster has had a significant impact on the reported literature. Although unrelated to VC, the cause of this cluster remains uncertain. PMID- 12762079 TI - A case-cohort study of angiosarcoma of the liver and brain cancer at a polymer production plant. AB - Past studies have reported an excess of angiosarcoma of the liver and brain cancer at a polymer production plant in Louisville, Kentucky. Although angiosarcoma has clearly been linked to vinyl chloride exposure, the cause of the brain cancer excess has been uncertain. We conducted case-cohort and case-control studies of both conditions in relation to chemicals used in polyvinyl chloride and nitrile rubber production. Angiosarcoma was strongly associated with vinyl chloride exposure but not with other chemicals used in production. Brain cancer was highest in workers hired prior to 1950 but was not associated with vinyl chloride or other specific exposures or production processes. This brain cancer cluster is similar to those reported in other industries in this generation of workers, the specific cause remaining unknown. PMID- 12762080 TI - Occurrence of lead-related symptoms below the current occupational safety and health act allowable blood lead levels. AB - To determine the occurrence of symptoms of lead toxicity at levels below the current allowable Occupational Safety and Health Act blood lead level of 50 micrograms/dL, standardized telephone interviews were conducted of individuals reported to a statewide laboratory-based surveillance system. Four hundred and ninety-seven, or 75%, of the eligible participants were interviewed. Gastrointestinal, musculoskeletal, and nervous system symptoms increased with increasing blood lead levels. Nervous, gastrointestinal, and musculoskeletal symptoms all began to be increased in individuals with blood leads between 30-39 micrograms/dL and possibly at levels as low as 25-30 micrograms/dL for nervous system symptoms. The results of this study of increased symptoms are consistent with and provide added weight to previous results showing subclinical changes in the neurologic and renal systems and sperm counts at blood lead levels currently allowed by the Occupational Safety and Health Act. PMID- 12762081 TI - Cleaning products and work-related asthma. AB - To describe the characteristics of individuals with work-related asthma associated with exposure to cleaning products, data from the California-, Massachusetts-, Michigan-, and New Jersey state-based surveillance systems of work-related asthma were used to identify cases of asthma associated with exposure to cleaning products at work. From 1993 to 1997, 236 (12%) of the 1915 confirmed cases of work-related asthma identified by the four states were associated with exposure to cleaning products. Eighty percent of the reports were of new-onset asthma and 20% were work-aggravated asthma. Among the new-onset cases, 22% were consistent with reactive airways dysfunction syndrome. Individuals identified were generally women (75%), white non-Hispanic (68%), and 45 years or older (64%). Their most likely exposure had been in medical settings (39%), schools (13%), or hotels (6%), and they were most likely to work as janitor/cleaners (22%), nurse/nurses' aides (20%), or clerical staff (13%). However, cases were reported with exposure to cleaning products across a wide range of job titles. Cleaning products contain a diverse group of chemicals that are used in a wide range of industries and occupations as well as in the home. Their potential to cause or aggravate asthma has recently been recognized. Further work to characterize the specific agents and the circumstances of their use associated with asthma is needed. Additional research to investigate the frequency of adverse respiratory effects among regular users, such as housekeeping staff, is also needed. In the interim, we recommend attention to adequate ventilation, improved warning labels and Material Safety Data Sheets, and workplace training and education. PMID- 12762082 TI - Mortality of mental disorders in relation to potential pesticide exposure. AB - Some studies have suggested a role of pesticide exposure in the development of neurobehavioral disorders. This case-control study examined the association between mortality from mental disorders and occupational exposure to pesticides. The study population consisted of 7756 deaths and 330,452 eligible controls identified from US death certificate files for the years 1988 through 1992. Exposure assignment was based on job title reported on the death certificates. Employment in jobs potentially involving pesticide exposure was weakly associated with the risk of death from mental disorders (odds ratio [OR] = 1.46; 95% confidence interval (95% CI) = 1.33-1.60). This association was stronger among women (OR = 2.65; 95% CI = 1.89-3.71), in particular for deaths from neurotic disorders (OR = 4.32; 95% CI = 2.44-7.64). These results must be interpreted with caution, however, because the impact of social and work-related factors other than pesticide exposure is not known. PMID- 12762083 TI - A pilot study of telephone-based smoking cessation intervention in asbestos workers. AB - Smoking markedly increases the risk of asbestos-related lung cancer. We conducted a randomized pilot trial of a telephone-based smoking cessation intervention in asbestos workers. Fifty-nine smokers were assigned to either a control or telephone-based smoking cessation treatment group and were followed-up at 6 months. Intent-to-treat analysis revealed a 16.7% quit rate at 6 months for the intervention group compared to 6.9% for the control group (P = 0.25). Treatment received quit-rates were 33% for the intervention group and 6.9% for the control group (P = 0.05). The intervention group was twice as likely to use smoking cessation medicines and progressed further along the stage of change continuum compared with the control group. Incorporating telephone-based smoking cessation treatment into medical screening activities for asbestos workers is feasible and the intervention is effective in increasing quit rates at 6 months. PMID- 12762084 TI - Community environment and women's health outcomes: contextual data. AB - OBJECTIVES: This report presents some illustrative data and analyses from the Contextual Data File for the 1995 National Survey of Family Growth (NSFG). Data are shown by the woman's race and Hispanic origin, and selected characteristics of the community in which she lived. METHODS: Cycle 5 of the NSFG was based on in person interviews with a national sample of 10,847 women 15-44 years of age in the United States in 1995. The interview included questions on the woman's births, marriages, contraceptive use, and characteristics such as her race and education. Measures of the characteristics of the woman's neighborhood were added to the interview data. RESULTS: This report shows that several simple measures of the social and economic status (SES) and resources of the woman's community of residence are closely associated with outcomes such as delayed childbearing, unwanted births, current marital status, the use of male or female contraceptive sterilization, breast-feeding, vaginal douching, and cigarette smoking. CONCLUSIONS: It is well-documented that the outcomes studied in this report are closely associated with individual characteristics such as age, race, education, and household income. But this report shows that these outcomes are also related to characteristics of the communities in which the individuals live. Researchers are encouraged to use the NSFG Contextual Data File to study these relationships further. PMID- 12762085 TI - Epidemiologic measures of the course and outcome of pregnancy. PMID- 12762086 TI - Bacterial vaginosis in pregnancy: current findings and future directions. PMID- 12762089 TI - Heritable and nonheritable risk factors for autism spectrum disorders. PMID- 12762088 TI - Evolution of surveillance of measles, mumps, and rubella in England and Wales: providing the platform for evidence-based vaccination policy. PMID- 12762090 TI - Environmental epidemiology of pediatric asthma and allergy. PMID- 12762091 TI - Genetic and perinatal risk factors for asthma onset and severity: a review and theoretical analysis. PMID- 12762092 TI - Relation between elevated ambient temperature and mortality: a review of the epidemiologic evidence. PMID- 12762087 TI - Vaginal douching: evidence for risks or benefits to women's health. PMID- 12762093 TI - Do indoor molds in nonindustrial environments threaten workers' health? A review of the epidemiologic evidence. PMID- 12762094 TI - Prevalence of symptoms and symptom-based conditions among Gulf War veterans: current status of research findings. PMID- 12762095 TI - Prevention of lower extremity stress fractures in athletes and soldiers: a systematic review. PMID- 12762096 TI - Pharmacologic agents associated with a preventive effect on Alzheimer's disease: a review of the epidemiologic evidence. PMID- 12762097 TI - Blood transfusions and non-Hodgkin's lymphoma. PMID- 12762098 TI - Failure of equipoise to resolve the ethical tension in a randomized clinical trial. PMID- 12762099 TI - Providing relief to those in pain: a retrospective on the scholarship and impact of the Mayday Project. PMID- 12762100 TI - Achieving the right balance in oversight of physician opioid prescribing for pain: the role of state medical boards. PMID- 12762101 TI - Maximizing the value of electronic prescription monitoring programs. PMID- 12762103 TI - Pain relief, prescription drugs, and prosecution: a four-state survey of chief prosecutors. PMID- 12762102 TI - Challenges in the federal regulation of pain management technologies. PMID- 12762104 TI - Monitoring and investigating certified registered nurse practitioners in pain management. PMID- 12762105 TI - Improving state medical board policies: influence of a model. PMID- 12762106 TI - A comparative study of the law of palliative care and end-of-life treatment. PMID- 12762107 TI - AIDS and antiretroviral drugs in South Africa: public health, politics, and individual suffering: a review of Brian Tilley's It's my life. PMID- 12762108 TI - The invisible vulnerable: the economically and educationally disadvantaged subjects of clinical research. PMID- 12762109 TI - Criminal law: physician convicted for recklessly prescribing OxyContin. PMID- 12762110 TI - Prenatal care: revisions to SCHIP extend health care to "unborn children". PMID- 12762111 TI - Managed care: health providers' bill of rights now law in California. PMID- 12762112 TI - Medicare: Ninth Circuit limits rates providers can charge Medigap insurers. PMID- 12762113 TI - Managed care: immunity for peer review under HCQIA. PMID- 12762114 TI - Tobacco litigation: statistics permitted for proof of causation and damages in class action. PMID- 12762115 TI - Public health: smoking ban exceeds Board of Health's authority. PMID- 12762116 TI - ADA: isolated bouts of depression do not qualify as a disability. PMID- 12762117 TI - Employment: protecting public health abrogates due process requirement for suspension proceedings. PMID- 12762118 TI - EMTALA: screening can satisfy EMTALA, despite misdiagnosis. PMID- 12762119 TI - Growth and survival of selected pathogens in margarine-style table spreads. AB - Although margarine-style table spreads can have a pH above 4.6 and a water activity greater than 0.85, there is some question if such products can support the growth of pathogenic bacteria. The objective of this study was to evaluate the growth and survival of Staphylococcus aureus, Listeria monocytogenes, Escherichia coli O157:H7, and Salmonella typhi in 60-percent- and 70-percent vegetable-oil, margarine-style, water-in-oil-emulsion table spreads stored at different temperatures. Samples of 25 grams of each table spread were inoculated with 1 x 10(3) cells of each bacterial mixture. The samples were stored at 5 degrees C, 7 degrees C, and 21 degrees C, and the microbial population in colony forming units per gram (CFU/gram) was enumerated over time. In almost all storage conditions, bacterial levels were shown to decrease over time. Inactivation was observed in (listed from fastest to slowest, respectively) S. aureus, L. monocytogenes, E. coli O157:H7, and S. typhi. Growth was observed only for S. typhi in table spreads stored at 21 degrees C, but the rate of growth was extremely slow. Based on these findings, the table spreads evaluated in this study are not potentially hazardous foods, and cold temperature storage is not necessary from a food safety perspective. PMID- 12762121 TI - Campylobacter jejuni enteritis associated with consumption of raw milk. AB - An outbreak of Campylobacter jejuni enteritis occurred among people who had attended a meal where raw milk was served. A case control study was conducted using instances of illness as cases; those who attended the event but did not become ill served as controls. Thirteen of 20 people who had attended the meal became ill. C. jejuni was cultured from five of six stools that were submitted. Raw milk consumption was strongly associated with illness (p = .0072, Fisher exact test). Although C. jejuni outbreaks associated with milk can be prevented with pasteurization, they still occur in association with raw milk consumption. PMID- 12762120 TI - A common-source outbreak of trichinosis from consumption of bear meat. AB - This paper discusses an outbreak of trichinosis that occurred in 1998 in Montgomery County, Ohio, and the investigation that followed. The outbreak was associated with consumption of bear meat from a hunt in Ontario, Canada. The person who had the index case had eaten two bear burgers that were cooked rare in a microwave oven. Bear meat from the same hunt later was consumed by 15 other people at a church supper and an additional 13 people who did not attend the supper. Of the 15 attendees at the church supper who ate the bear meat, seven developed illness consistent with Trichinella infection (attack rate about 47 percent). An additional seven people attended the supper but did not eat the bear meat and did not become ill. Having eaten bear meat at the church supper was associated with an increased risk of illness (p = .05). Inadequate cooking of the bear meat resulted in the transmission of live trichinae. The 13 other people who ate the bear meat but did not attend the supper reported no illness. A total of eight people, including the person with the index case, met the case definition for trichinosis. Adequate cooking of the bear meat or consumption of uninfected portions of the meat was probably the protective factor for those who did not become ill after consuming the bear meat. PMID- 12762122 TI - The environmental health professionals of the Volusia County Health Department: a focus on charity. PMID- 12762123 TI - Health department liability for non-enforcement. PMID- 12762124 TI - The customer is always right. PMID- 12762125 TI - Illegal meth labs: cleanup raises health and environmental concerns. PMID- 12762127 TI - Multispecialty group transforms itself as it turns to capitation. PMID- 12762126 TI - Botulism--information from the World Health Organization. PMID- 12762128 TI - IDS bolts from capitation contract, citing low rates and unfair contract provisions. PMID- 12762129 TI - Physician dissatisfaction: how do doctors under capitation fare? PMID- 12762130 TI - Study shows savings arise from spending on medical management. PMID- 12762131 TI - ETGs offer capitated groups multiple options. PMID- 12762132 TI - Pre-release. Tuberculosis in Canada, 2001. PMID- 12762133 TI - Contact investigation of a case of pulmonary and laryngeal tuberculosis. PMID- 12762135 TI - An allele of serum amyloid A1 associated with amyloidosis in both Japanese and Caucasians. AB - Serum amyloid A1 (SAA1), one of the two isotypes of acute phase SAA, is the predominant precursor to amyloid A (AA) protein, the chief constituent of fibrillar deposits in reactive (AA) amyloidosis. Prolonged hyperexpression of SAA protein accompanying chronic inflammation is critical to, but seems not to be sufficient for, the development of AA amyloidosis. Several previous studies have investigated the possibility of linkage between SAA1 exon 3 polymorphisms and susceptibility to amyloidosis. While the SAA1.1 allele was found to have a negative association with amylodosis in Japanese subjects, it showed a positive association in Caucasians. Moriguchi and colleagues recently showed that a single nucleotide polymorphism (SNP) at position -13 in the SAA1 5' flanking region was more strongly associated with amyloidosis than was the exon 3 polymorphism. To test whether this SNP may be an amyloidogenic factor common to Japanese and Caucasians, we have analyzed the SAA1 gene in amyloid and non-amyloid patients of both ethnic groups for the presence of T or C at position -13 and for exon 3 polymorphisms (SAA1.1, 1.3 or 1.5). The frequency of the -13T allele was 0.708 and 0.521 in Japanese rheumatoid arthritis patients with and patients without AA amyloidosis, respectively, and 0.536 and 0.196 in American Caucasian patients with AA amyloidosis and control subjects, respectively. In Caucasians, the -13T allele had a stronger association with amyloidosis than did the SAA1.1 allele. These findings suggest that -13T is a genetic background for AA amyloidosis in both Japanese and Caucasians and the difference in prevalence of AA amyloidosis in the two ethnic groups may be due, at least in part, to a difference in the frequency of the -13T SAA1 allele. PMID- 12762134 TI - Treatment with the selective muscarinic m1 agonist talsaclidine decreases cerebrospinal fluid levels of A beta 42 in patients with Alzheimer's disease. AB - The amyloid beta-peptides A beta 40 and A beta 42 are highly amyloidogenic constituents of brain beta-amyloid plaques in Alzheimer's disease (AD). Lowering their formation may be achieved by modulating the activities of proteases that cleave the amyloid precursor protein (A beta PP), including alpha- beta-, and gamma-secretases. Talsaclidine is a functionally selective muscarinic m1 agonist that stimulates non-amyloidogenic alpha-secretase processing in vitro. We compared cerebrospinal fluid (CSF) levels of A beta 40 and A beta 42 measured by ELISA before and at the end of 4 weeks of treatment with talsaclidine. The medication was administered in a double-blind, placebo-controlled, and randomized clinical study to 40 patients with AD. Talsaclidine (n = 34) decreased CSF levels of A beta 42 by a median of 19% (p < 0.001) as compared to baseline. The mean difference between CSF levels of A beta 42 before and after treatment with talsaclidine (n = 34) was -46 +/- 73 (SD) pg/ml as compared to 0 +/- 8 (SD) pg/ml with placebo (n = 6) (p < 0.05). CSF levels of A beta 40 increased during treatment with placebo (n = 6) while they remained stable during treatment with talsaclidine (n = 31) (1.118 +/- 1.710 ng/ml, and -0.170 +/- 0.967 ng/ml, respectively; p < 0.05). These data show that treatment with the m1 agonist talsaclidine reduced A beta peptides, and particularly A beta 42, in AD patients, suggesting it as a potential amyloid lowering therapy of AD. PMID- 12762136 TI - Serum amyloid A1 and tumor necrosis factor-alpha alleles in Turkish familial Mediterranean fever patients with and without amyloidosis. AB - The major complication of familial Mediterranean fever (FMF) is AA amyloidosis. The influence of FMF gene (MEFV) mutations and/or unknown environmental factors and other genetic modifiers are likely to affect the phenotypic variations of the disease and the development of amyloidosis. Serum amyloid A is a serum precursor of AA amyloid that is induced by inflammatory cytokines including TNF-alpha. Our analysis of SAA1.1 frequency in Turkish FMF-amyloidosis patients, revealed a higher frequency compared to non FMF-amyloidosis patients but the difference was not significant. On the other hand, the distribution of SAA1.1 homozygosity among FMF-amyloidosis patients was 55.5% compared to FMF-non-amyloidosis patients (30.8%) which was statistically significant revealing a 2.5 fold risk for the occurrence of amyloidosis. There was no significant difference between the controls and FMF patients with and without amyloidosis for the TNF-alpha-308 G-A allele. It is worth noting that all TNF-alpha-308 G-A carriers (n = 6) in FMF amyloidosis group have SAA1.1 homozygosity compared to 2/11 in FMF-non amyloidosis group. Further evaluation of these polymorphisms may have importance and need further study. PMID- 12762137 TI - Peripheral nerve function in patients with familial amyloid polyneuropathy after liver transplantation. AB - To evaluate the therapeutic efficacy of liver transplantation in patients with ATTR Val30Met familial amyloid polyneuropathy (FAP), were repeatedly examined the neurophysiological function of peripheral nerves in nine patients. The maximal motor and sensory conduction velocities (MCV and SCV) of the ulnar and tibial nerves, size of compound muscle action potential (CMAP), terminal latency of CMAP, skin temperature of extremities, CVR-R, blood pressure, heart rate, and Schellong's test were examined before and every 6 months after the operation. Although there were no changes in CVR-R, blood pressure, or heart rate, the skin temperature of foot and hand increased soon after surgery and did not decrease during the period of observation. The temperature-adjusted MCV of tibial nerve gradually increased, but the MCV of ulnar nerve showed no change. The temperature adjusted tibial nerve SCV worsened slightly soon after transplantation and remained at that level in the distal part. The ulnar nerve SCV worsened and subsequently improved. Liver transplantation is very effective for halting the progression of this type of FAP, but the recovery of peripheral nerve function in patients seems to be very slow and limited, especially the function of large diameter myelinated fibers. PMID- 12762138 TI - Transthyretin Tyr69-to-Ile mutation (double-nucleotide substitution in codon 69) in a Japanese familial amyloidosis patient with cardiomyopathy and carpal tunnel syndrome. AB - A 61-year-old Japanese woman with transthyretin amyloid (ATTR) Tyr69Ile, which was caused by the mutation of TTR gene TAC to ATC at codon 69, is described. The patient had no family history and developed carpal tunnel syndrome followed by congestive heart failure due to cardiac amyloidosis. Various phenotypes of familial transthyretin amyloidosis including FAP are caused by TTR variants with single amino-acid substitutions, the latter being caused by one-point mutations in the coding region of the TTR gene. This is the first report showing a novel double-nucleotide substitution in the causative TTR gene abnormality. PMID- 12762139 TI - A transthyretin mutation (Tyr78Phe) associated with peripheral neuropathy, carpal tunnel syndrome and skin amyloidosis. AB - BACKGROUND: More than 80 transthyretin (TTR) mutations have been described, most associated with amyloidosis. Peripheral neuropathy is the most common clinical presentation in TTR amyloidosis although the carpal tunnel syndrome (CTS) may be the first symptom and skin can be involved, as transthyretin amyloidosis is a systemic disease. CASE REPORT: The 78 year-old proband, belonging to a French family of Italian origin, presented with a 5 year history of peripheral neuropathy in the lower extremities. However, 15 years earlier he had had surgery for bilateral CTS. Amyloidosis was diagnosed on salivary gland and skin biopsies. Immunohistochemistry on skin biopsy was positive using anti-TTR. The proband has 10 siblings, 5 have CTS. METHODS: SSCP and direct sequencing of exons 2, 3, and 4 of the TTR gene were done on DNA from the proband and his brother who had had CTS. To confirm the mutation a PCR-IMRA was done. RESULTS: SSCP analysis of TTR exons 2, 3, and 4 did not suggest a mutation. Sequence analysis of TTR exon 3 revealed heterozygosity in both subjects for a single basepair transversion from A to T in codon 78 (TAC-->TTC) indicating a tyrosine to phenylalanine change. The mutation was confirmed by PCR-IMRA. CONCLUSION: This TTR mutation (Tyr78Phe) is associated with peripheral neuropathy, carpal tunnel syndrome and skin amyloidosis. It is also associated with late onset of the disease. PMID- 12762140 TI - First Spanish family with familial amyloidotic polyneuropathy associated to TTR Thr49Ile mutation. AB - We present a Spanish patient with familial amyloidotic polyneuropathy associated with the TTR Thr49Ile mutation previously described in a Japanese patient. This is the first report in a Caucasian patient and the second in the literature. Age of onset at 66 and the clinical picture were similar to the Japanese patient: sensorimotor polyneuropathy, digestive autonomic disturbances, cardiomyopathy and loss of weight. The mutation was diagnosed by DNA sequencing and induced mutation restriction analysis. PMID- 12762141 TI - Localized AL amyloidosis of the colon: an unrecognized entity. AB - Virtually all patients who present with rectal bleeding and amyloid of the colon have evidence of systemic amyloidosis and require therapy. The small subset of patients with amyloidosis localized to the colon must be recognized and treatment avoided. We queried our file for patients who had amyloidosis of the colon but no evidence of systemic amyloidosis during long-term follow-up. We identified 3 patients who presented with rectal bleeding and who, on investigation, had primary amyloidosis of the colon but no evidence of systemic amyloidosis during a follow-up of 4.5 to 20 years. These patients had no evidence of a plasma cell dyscrasia and received no chemotherapy to prevent deposition of amyloid. It is important to recognize this rare subset and avoid treatment with alkylating agents or high-dose therapy followed by autologous stem cell transplantation. Alkylating agent therapy may be associated with myelodysplasia or acute leukemia. In addition, the cost, inconvenience, and morbidity of therapy are avoided by observation. Patients who present with rectal bleeding and a subsequent diagnosis of amyloidosis of the colon likely will be subjected to chemotherapy or transplantation. Such patients must be recognized and treatment avoided if there is no evidence of systemic amyloidosis because they remain stable for many years. PMID- 12762142 TI - Spontaneous pneumoperitoneum: an unusual complication of systemic reactive AA amyloidosis secondary to rheumatoid arthritis. AB - We report a 71-year-old man with reactive AA amyloidosis secondary to rheumatoid arthritis who developed spontaneous pneumoperitoneum with intestinal pseudo obstruction as an initial symptom. Severe deposition of amyloid in the intestinal wall was considered to play an important role in the pathogenesis of this unusual symptom. The patient has been successfully treated with total parenteral alimentation and intermediate-dose prednisolone (30 mg/day). Although pneumoperitoneum usually suggests gastrointestinal perforation requiring emergency surgery, conservative therapy should be seriously considered in amyloidosis-related cases with no associated peritonitis, since multiple vital organs are probably involved by severe amyloid deposition, thus increasing the risks of surgery. PMID- 12762143 TI - Localized epidural and bone amyloidosis, rare cause of paraplegia in multiple myeloma. AB - We report two cases of spinal cord compression by AL amyloid deposits in the setting of multiple myeloma. The first patient, a 57 year old man, had a surgical procedure late in his course, when his neurological status worsened despite medical treatment. The second patient, a 61 year old man, had surgical treatment as soon as vertebral body collapse and epiduritis were diagnosed and spinal amyloidosis revealed a non secretory myeloma. Neurological recovery was significant in both cases and neither developed manifestations of systemic amyloidosis 4 years later. PMID- 12762144 TI - The 8th International Conference on Alzheimer's Disease and Related Disorders, July 20-25, 2002, Stockholm, Sweden. PMID- 12762145 TI - [Current terminology for tooth wear]. AB - In general dental practice the "abrasion" diagnosis is universally used for the loss of any tooth structure irrespective of its etiology. The author's aim was to summarize and classify the types of non-caries-related tooth wear (i.e. attrition, abrasion and erosion) based on their etiology and clinical features. All dental practitioners should be aware of the criteria of differentiation and the principles of differential diagnoses because only the elimination of the etiologic factors can ensure permanent therapeutic effect. On he other hand dental practitioners should also be familiar with this field to be able to recognize the early symptoms of certain general diseases that can initially be diagnosed in he dental office. Besides the elimination of the causative factors other specialist should also be involved in the comprehensive treatment to provide long-term success and arrest the progression of the disease. PMID- 12762146 TI - [Effect of long-lasting clenching on dental occlusion]. AB - Durable clenching on a particular occlusal area may affect the masticatory muscle activity and induce transitional deformation of the dentoalveolar, mandibular and temporomandibular tissues. These effects may provide detectable alterations in the occlusal contact pattern. The aim of this study was to evaluate the subsequent effect of clenching on the number and location of occlusal contacts. This study was carried out on 16 volunteers with correct occlusion and sharply demarcated occlusal contact pattern. The patient closed to intercuspal position with maximal biting force and the occlusal contacts were revealed with a 12 microns thick occlusal foil. Then a gnathometer was placed between the upper and lower incisors and the mouth was closed for two minutes with about 50 N closing force. After that the occlusal contacts were evaluated again. Paired t-test was used to evaluate the data. Both before and after clenching the highest average number of contacts was recorded on the surface of first molars. They were followed in decreasing order by the second molars, second premolars, first premolars, and finally the canines. When the number of contacts was compared in tooth groups, the first molars and the second premolars had more contacts than the second molars and the first premolars. On the surfaces of molars more occlusal contacts were registered after clenching than before. Significant differences were found in case of the left second molar only. It was concluded that durable clenching on a particular occlusal area might change transitionally the distribution of occlusal contacts. PMID- 12762147 TI - [Study of the effects of alkali metals on some virulence characteristics of Candida albicans]. AB - The effects of the alkali metals sodium, potassium and lithium on the growth and on certain virulence factors (adhesion, cell-surface hydrophobicity and the germinating ability) of Candida albicans were investigated. It can be concluded that high concentrations of alkali metals possessed an inhibitory effect on the growth of the Candida cells and preincubation in the presence of alkali metals had a negative effect on all the virulence factors studied. It is worth emphasizing that the changes induced during the preincubation persisted even when the high concentrations of the alkali metals were removed from the cell suspension. However, even at high concentrations of sodium or potassium a considerable growth of Candida cells could be measured. Data also showed that although alkali metals could significantly decrease certain virulence traits of the fungus they could not totally inhibit either the adhesion or the germ tube formation potential of the cells. Thus, in spite of the high salt concentrations Candida cells may represent a health hazard in such habitats. PMID- 12762148 TI - [Prevalence of impacted permanent upper canine and its treatment in 11-18-year old orthodontic patients]. AB - Data of literature show very high prevalence of impacted upper permanent canine. The aim of the present study was to evaluate prevalence of retentioned or impacted upper permanent canines and to present their therapy. Data on 11-18-year old children presented for treatment at the Department of Dentistry for Children and Orthodontics in Budapest over a period of 10 years (between 1990 and 2000) were analysed. OP X-ray films of 1858 children and adolescents--1102 female and 756 male--were examined and the data of anamnesis and the administered therapy were considered. Results revealed 101 cases of impacted cuspids, the male:female ratio was 46:55. Considering the totality of administered treatment methods, both surgical exposure and orthodontic treatment of the affected canines were needed in a high rate, i.e. 96.92% of the analysed cases. In 40.91% of the cases extraction of the correspondent upper first premolar was needed. Following the premolar extraction spontaneous occlusions of the impacted canine was established in 3.03%. Authors present the complex therapy of one case of palatally impacted upper permanent canine. PMID- 12762149 TI - [Dental wear as a stomatologic manifestation of bulimia nervosa--a clinical case]. AB - The number of patients with anorexia and bulimia nervosa is increasing nowadays. The typical oral feature of these eating disorders is the dental erosion which causes sensitivity of the teeth and esthetic problems for the involved patients. This phenomenon is a characteristic feature in these cases and it may be the first sign of the mentioned disorders. The purpose of the study was to describe the generally the most characteristic oral findings of bulimia nervosa and anorexia nervosa because the dentists play a significant role in recognizing the basic problem of the patients, and they can send them for medical treatment of the serious general problems. PMID- 12762150 TI - [Dental journals in Transylvania published fully or partially in Hungarian, between 1919 and 1940]. AB - Between the two world wars several medical journals were published in Transylvania for shorter or longer periods of time fully or partly in Hungarian language. Only dentistry had its own periodicals, two for dentists (Revista Stomatologica--Stomatologiai Szemle 1823-1940, Stomatologia 1929-1937) and one for dental technicians (Viata Dentara--Dentistisches Fachblatt--Fogaszati Szaklap 1934). Because of the difficulties in their accessibility little or inaccurate data have been published so far on these journals. It is the main goal of the present paper to give an overview on these journals, which are very valuable not just for their professional merits but also for their historic value. PMID- 12762151 TI - [Statistical analysis of the "Dental Fear Questionnaire" surveying various subpopulations in Hungary]. AB - Authors investigated the Dental Fear Survey scores (DFS) of 362 persons from several subpopulations living in Budapest. Subjects were: 253 females, 109 males, aged 14 to 73 years. Scores of the items related to dental handpiece and anaesthetic needle were the highest. The group the participants belonged to influenced 19 items, marital status and age influenced 13 items, and sex influenced 10 items of the DFS significantly (one-way ANOVA, p < or = 0.05). Scores related to dental handpiece were higher than scores related to the anaesthetic needle, and the scores of the 20th item ("overall fear of dentistry") were near to the scores of most fearful items, indicating some "phobic character" of the Hungarian population. PMID- 12762152 TI - Pancreatic carcinoma. PMID- 12762153 TI - Cervical carcinoma clinical update. PMID- 12762154 TI - Prostate cancer: screening and treatment options. PMID- 12762155 TI - Plasma cell disorders: myeloma and related conditions. Diagnosis and management. PMID- 12762156 TI - Treatments for oral, pharyngeal and laryngeal carcinomas: a work in progress. PMID- 12762157 TI - Esophageal cancer. PMID- 12762158 TI - Thoughts on the doctor. PMID- 12762160 TI - Cancer. PMID- 12762161 TI - In a disaster, will your OR supply chain handle a wave of casualties? PMID- 12762162 TI - How building a 'just culture' helps an organization learn from errors. PMID- 12762163 TI - Are you confident about scope reprocessing? PMID- 12762164 TI - Hospital finds savings by outsourcing cataracts. PMID- 12762165 TI - A blame-free culture in the real world. PMID- 12762166 TI - ASC list a mix of wins, disappointments. PMID- 12762167 TI - Complying with HIPAA's security rule. PMID- 12762168 TI - Severe acute respiratory syndrome--Singapore, 2003. PMID- 12762169 TI - Buruli ulcer disease. Mycobacterium ulcerans infection. PMID- 12762170 TI - Accounting of disclosures. PMID- 12762171 TI - Current management of epithelial thyroid neoplasms. AB - Most patients with thyroid cancer have one of the epithelial carcinomas, either papillary or follicular. While most of these patients have favorable outcomes, an understanding of current methods to evaluate, treat, and follow these patients allows physicians to provide better care and identify those most at risk. PMID- 12762173 TI - Humility. PMID- 12762172 TI - A comparison of applicant and matriculant trends, and rising costs of medical education in United States medical schools and at the University of Kentucky College of Medicine. AB - This paper addresses fluctuations in the applicant and matriculant pools both across United States medical schools and at the University of Kentucky College of Medicine (UKCOM) for 1992-2002. It also presents data regarding the increasing costs of a medical education. Over the past decade, both nationally and at the UKCOM, there has been an over-all reduction in the number of applicants to medical school. In this changing applicant pool, the percentage of female matriculants has increased both nationally and at the UKCOM. However, the number of underrepresented minorities applying to and matriculating in the US and at the UKCOM has dropped since the mid-1990s. Although the applicant pool has decreased in size over the time period examined, the academic quality of applicants as measured by the undergraduate grade point average and Medical College Admission Test scores has increased both nationally and at UKCOM. Costs of a medical education have risen over time, as has the debt burden of medical school graduates due to increasing undergraduate debt, consumer debt, and medical school tuition. Potential causes for and implication of these changing trends are discussed. PMID- 12762174 TI - Affective review and schema reliance in memory in older and younger adults. AB - Previous studies have found that, when remembering, older adults often rely more on schematic knowledge than do younger adults. We replicated this finding when participants were induced to review the facts of the event; furthermore, neuropsychological correlates suggested that this age-related increase in schema reliance is associated with declines in reflective processes. In addition, when they were induced to focus on their feelings and reactions when reviewing an event, both older and younger adults' later memory of the event was strongly affected by their schematic knowledge. PMID- 12762175 TI - Psycholinguistics: a half century of monologism. AB - The contention of this article is that, since its inception in the mid-twentieth century, mainstream psycholinguistics has been monologistic, that is, has concentrated on monologue as its source of empirical material and has, largely implicitly, involved a monologistic epistemology. The article is not a comprehensive history of psycholinguistics but does attempt to establish a historical perspective. Monologism has been the historical bias of Cartesianism, positivism, behaviorism, and cognitivism. Monologism is concerned only with the person in whom cognition takes place and from whom communication proceeds. It is essentially asocial. By contrast, the merits of dialogism include an openness to the sociocultural, interactive nature of all cognition and communication and an empirical engagement of contextualized discourse situations. Dialogism is presented here not as a supplement to mainstream psycholinguistics but as a radical innovation that construes mainstream psycholinguistics as "strongly misleading if presented as a full theory of communication through spoken interaction" (Linell, 1998, p. 23). Some approaches to dialogism and research on dialogue are critically reviewed. PMID- 12762176 TI - A paradoxical dissociation in the effects of midazolam on recollection and automatic processes in the process dissociation procedure. AB - This study used midazolam-induced amnesia to explore the plausibility of the estimates provided by the process dissociation procedure (PDP), which is designed to estimate the contributions of recollection (R) and automatic (A) processes to implicit memory performance. In a double-blind, placebo-controlled, within subject design with 24 participants, single midazolam doses were administered intravenously, and word stem completion performance was used to calculate PDP estimates. A dissociation was observed such that midazolam decreased R but increased A estimates relative to placebo. Given that a manipulation that induces amnesia would not be expected to facilitate a memory process, these results add to the accumulating body of evidence suggesting that PDP estimates are not always theoretically plausible. Such evidence raises important questions about the use of the PDP. PMID- 12762177 TI - Implicit temporal tuning of working memory strategy during cognitive skill acquisition. AB - Complex cognitive tasks such as multiple-step arithmetic entail strategies for coordinating mental processes such as calculation with processes for managing working memory (WM). Such strategies must be sensitive to factors such as the time needed for calculation. In 2 experiments we tested whether people can learn the timing constraints on WM demands when those constraints are implicitly imposed. We varied the retention period for intermediate results using the well known digit size effect: The larger the operands, the longer it takes to perform addition. During learning participants practiced multiple-step arithmetic routines combined with large or small digits. At transfer, they performed both practiced and novel combinations. Practice performance was affected by digit size and WM demands. However, the transfer performance was not fully explained by the digit size effect or the practice effect. We argue that participants acquired temporal tuning of the WM strategy to the implicit retention interval imposed by the digit size and kept using the tuning mode to unpracticed data set. PMID- 12762178 TI - Study-test awareness can enhance priming on an implicit memory task: evidence from a word completion task. AB - The role of study-test awareness in implicit memory tasks has been an open question for some time. This study investigated the possibility that study-test awareness may enhance priming on an implicit memory task. In three experiments, subjects studied words under levels of processing conditions (nonsemantic vs. semantic) and then received a word stem completion priming task. The results of all three experiments showed that study-test awareness had no effect on priming for nonsemantic study but it did on semantic study, significantly enhancing priming for that study condition. The results are interpreted according to an involuntary aware memory framework. PMID- 12762179 TI - Involuntary aware memory enhances priming on a conceptual implicit memory task. AB - This study investigated the role of involuntary aware memory (phenomenological awareness of the study episode) on a conceptual implicit memory task. Subjects studied words under levels of processing conditions (LOP: nonsemantic vs. semantic) and then received a category exemplar generation priming task. Subjects were either aware or unaware of the study test relationship. Subjects who were study test aware showed more priming for semantic study than subjects who were study test unaware, thus producing an LOP dissociation between them. The findings suggest that involuntary aware memory can enhance performance on conceptual implicit memory tasks, thus having theoretical implications for implicit and explicit memory. PMID- 12762180 TI - Incentive program focuses physicians on improving outcomes. PMID- 12762181 TI - Performance improvement at the heart of new cardiovascular center. PMID- 12762182 TI - Smallpox clinical pathway prepares Stanford for possible attack. PMID- 12762183 TI - Hospital patient experience surveys on track--are you ready? PMID- 12762184 TI - Access to hospice care. Expanding boundaries, overcoming barriers. PMID- 12762185 TI - Caregiving at the end of life. PMID- 12762186 TI - Health care justice and hospice care. PMID- 12762188 TI - What the people would want if they knew more about it: a case for the social marketing of hospice care. PMID- 12762187 TI - Pictures of persons and the good of hospice care. PMID- 12762189 TI - Hospice and Alzheimer disease: a study in access and simple justice. PMID- 12762190 TI - The relevance of public health in improving access to end of life care. PMID- 12762191 TI - Rediscovering community at the core of the human condition and social covenant. PMID- 12762193 TI - The complex path to managing mental illness. PMID- 12762192 TI - Is discontinuity in palliative care a culpable act of omission? PMID- 12762194 TI - Tackling foreign-worker recruitment. PMID- 12762195 TI - Character should be nurtured. PMID- 12762196 TI - [Risk factors of sudden infant death syndrome]. PMID- 12762197 TI - [Bacteriophage therapy]. PMID- 12762198 TI - [Risks of the cesarean section]. PMID- 12762199 TI - [Physical exercise is an advantageous and effective way to prevent falls and injuries due to falls]. PMID- 12762200 TI - [Qualitative health research--what, how and why?]. PMID- 12762201 TI - [Drug therapy and nutrition therapy of a young patient with Crohn disease]. PMID- 12762202 TI - [Cases of oculopharyngeal muscular dystrophy even in Finland]. PMID- 12762203 TI - [Postoperative atrial fibrillation]. PMID- 12762204 TI - [Bothersome nodules on the fingers of a musician]. PMID- 12762205 TI - [Brain injuries in adults]. PMID- 12762206 TI - Exploring aphasic grammar. 1: A single case analysis of conversation. AB - This paper uses the methodology and analytical findings of conversation analysis to investigate the notion that aphasic grammar may be understood at least partly in the context of the demands of turns at talk in conversation. An investigation of the conversation of an English-speaking person with aphasia reveals two distinct grammatical phenomena, and it is suggested that their use may be interactionally motivated by the need to take a relatively unproblematic turn at talk despite the constraints of non-fluent aphasia. The grammatical patterns that are revealed by this analysis look considerably different from those elicited by standard methods of data sampling. The possibility that interactional grammatical phenomena are not visible in the language data elicited by clinical assessments is raised. This question is addressed via an in-depth comparison of the same speaker's interactional and elicited grammar in a linked article. It is suggested that the tool of conversation analysis provides researchers with a new and fruitful approach to the study of grammatical abilities in aphasia. PMID- 12762207 TI - Exploring aphasic grammar. 2: Do language testing and conversation tell a similar story? AB - This paper investigates the grammatical difficulties of an English-speaking person with non-fluent aphasia using clinical assessments based on picture description and story telling. The same individual's conversation grammar, which was investigated in detail in a linked article is reviewed here, and the notion that interactional grammatical phenomena may not necessarily be visible in elicited language data is explored. Data analysis shows that the aphasic speaker's grammar looks considerably different in the context of clinical assessment than it does in conversation. Analysis of elicited grammar reveals that the majority of utterances produced are subject-verb-object (SVO) type sentences, whereas prior analysis of conversation data indicated patterns of interactional grammatical phenomena that differ from such sentence types. It is suggested that language tests and conversation provide complementary but essentially different information about grammatical abilities. The clinical implications of this finding are discussed. PMID- 12762208 TI - 'SLI', a generic category of language impairment that emerges from specific differences: a case study of two individual linguistic profiles. AB - This paper looks at data taken from two school-aged children labelled as 'SLI' and examines their individual linguistic profiles both at the outset of a specific therapy intervention and approximately 1 year later. The study sought to assess their language abilities within a conversational context and to assess the efficacy of a particular area of their therapy programme. Differences in the two children's individual LARSP profiles raised questions relating to (a) the overall usefulness of 'SLI' as a diagnostic category of disorder, and (b) the need for greater use of detailed syntactic analysis by Speech Language Pathologists in both diagnostics and in implementing therapy programmes. PMID- 12762209 TI - The omission of subject arguments in children with specific language impairment. AB - Ten children with specific language impairment (SLI) were compared to two groups of normally developing children for the production of grammatical subjects in sentences of varying length and argument structure complexity. The normal controls consisted of a group of younger children matched for mean length of utterance (MLU) and a group matched for chronological age. The participants were asked to produce sentences of varied argument structure complexity using a story completion task. The results indicated that both the children with SLI and the MLU controls omitted more subject arguments in the ditransitive sentences than in sentences with intransitive and ditransitive verbs. In addition, more children with SLI omitted subject arguments as linguistic complexity increased. This effect was not found for the normal age control children who never omitted subjects, regardless of increases in argument structure complexity. These results support the notion that grammatical errors in both children with SLI and their younger, normal counterparts may be due to problems with processing complex linguistic information rather than with limitations in linguistic knowledge. PMID- 12762210 TI - Thanks for HB 2122; now let's enact federal medical liability reform, fix Medicare. PMID- 12762211 TI - Benefits outweight risks when prescribing antibiotics. PMID- 12762212 TI - Stent-supported angioplasty correction of symptomatic critical carotid angulation. AB - Carotid stent-supported angioplasty is currently under investigation in many medical centers, for use in treating extracanial cerebrovascular disease. The early results of CSSA in selected patients appear promising. While carotid endarterectomy (CEA) remains the current standard of care, we believe that a small subgroup of patients at a high risk for surgery can benefit from CSSA. This case report describes a patient with symptomatic high-grade recurrent stenosis due to critical angulation (kinking) and redundancy of the internal carotid artery following CEA with patch angioplasty who was then treated successfully with CSSA. PMID- 12762213 TI - Persistent candiduria complicating intraureteral stenting: a case report and review of literature. AB - Over the past 20 years, there has been a significant increase in the number of nosocomial infections. Nosocomial infections by fungal organisms, in particular Candida species, have shown the highest increase in incidence, currently being the leading pathogen in urinary tract infections and the fourth most common blood born pathogen within hospitalized patients. This case report describes a patient with acute pancreatitis, complicated by acute renal failure, who developed nosocomial fungiuria and fungemia following bilateral ureteral stenting, and whose fungiuria persisted despite adequate antifungal treatment. Conditions resolved with removal of the intraureteral stents, which we believe, served as the nidus for her persistent fungiuria. PMID- 12762214 TI - Primary amyloidosis (AL) presenting with nephrotic syndrome: a case report and discussion. AB - Primary amyloidosis (AL) is a rare disorder with only eight cases per million a year. AL is a plasma cell disorder in which neoplastic clonal plasma cells in the bone marrow produce monoclonal immunoglobulin light chains that form protease resistant amyloid fibrils. AL fibrils accumulate within tissues systemically, causing progressive organ impairment to ultimate death. Median survival in AL with treatment is only 17 months depending upon variable prognostic factors. Most patients present with nephrotic syndrome and associated peripheral edema because the kidney is the most common organ involved in AL. This article describes the case of a patient who presented to City Hospital in Martinsburg, W.Va., and was diagnosed with primary amyloidosis, and reviews the current literature on this illness. PMID- 12762215 TI - Successful embolization of retroperitoneal bleeding from a renal angiomyolipoma. AB - The finding of acute flank pain, hypotension and anemia in the absence of trauma raises concerns about retroperitoneal bleeding. This article highlights the rare case of a patient with a bleeding retroperitoneal angiomyolipoma. PMID- 12762216 TI - New drug buprenorphine can treat patients addicted to heroin or prescription pain medications. PMID- 12762217 TI - [Problems of current antidepressant drugs]. AB - Currently, 17 antidepressant drugs are available in Japan. They are certainly evaluated as effective drugs based on abundant clinical evidence. However, treatment outcome of mood disorders does not seem to tremendously advance despite the growing number of antidepressant drugs available in clinical practice. The current antidepressant drugs, including SSRIs and SNRI, have common shortcomings from several clinical standpoints, such as 1) efficacy rate remains at 60-70%, 2) lack of immediate onset of efficacy, 3) side effects, 4) no pleasant effect, and 5) difficult prediction of efficacy for each drug. These problems are largely due to a fact that their prototypes are common. New strategy for development of antidepressants is expected. PMID- 12762218 TI - [Pharmacogenomics and antidepression research]. AB - Although antidepressants have been used clinically for more than 50 years, no consensus has been reached concerning their precise molecular mechanism of action. Pharmacogenomics is a powerful tool that can be used to identify genes affected by antidepressants or by other effective therapeutic manipulations. Using this tool we have previously identified hundreds of cDNA fragments as antidepressant related genes (ADRGs). Some of these candidate genes may encode common functional molecules induced by chronic antidepressant treatment. Defining the roles of these molecules in drug-induced neural plasticity is likely to transform the course of research on the biological basis of antidepressants. Such detailed knowledge will have profound effects on the diagnosis, prevention, and treatment of depression. Novel biological approaches beyond the "monoamine hypothesis" are expected to evoke paradigm shifts in the future of antidepressant research. PMID- 12762219 TI - [The approaches in the discovery of antidepressants using affective disorder models]. AB - With the recent appearance of SSRI and SNRI, medication options with respect to depression have broadened. However, patients displaying clear improvement with existing antidepressants still do not exceed about 60 percent of total patients. New types of therapeutic agent development are currently required. Conditions for the determination of new antidepressants are: 1) instantaneous medications displaying a high level of antidepressant action in the early stages of treatment and 2) medications displaying efficacy with respect to patients that are therapy resistant. However, drug discovery using new animal models is critical as part of drug development of these types of antidepressants in addition to models used in the past such as the forced swim test. We adopted two animal models (olfactory bulbectomy model and conditioned fear stress (CFS) model) developed for pharmacological evaluation of antidepressants. It has been well known that olfactory bulbectomied rats display extreme emotional response (aggressiveness and anxiety). However the improvement of this response occurs following chronic but not acute antidepressant treatment. Thus, we used this model to evaluate the period of the manifestation of antidepressant action. Mice exhibited a marked suppression of motility when they were returned to the same environment in which they had previously received an electric foot shock. Thus, it is suggested that the CFS stress model may be a useful model for therapy-resistant depression due to the fact that motor suppression is not readily attenuated by antidepressant treatment. In this report, we provide an overview of the approaches in the discovery of new antidepressants using these affective disorder models. PMID- 12762220 TI - [Possible involvement of GABAB receptors in action of antidepressants]. AB - Antidepressants are divided into several groups such as monoamine oxidase inhibitors, serotonin reuptake inhibitors, noradrenaline reuptake inhibitors and alpha-adrenoceptor antagonists. Antidepressants show, however, a common clinical profile: their clinical actions occur after a chronic treatment. It is, therefore, suggested that antidepressants have a common pharmacological profile. We reviewed possible involvement of gamma-aminobutyric acid (GABA)B receptors in action of antidepressants. Our data in combination with previous ones from other laboratories suggest that antidepressants suppress the activity of presynaptic 5 hydroxytryptamine3 receptors on GABA neurons, resulting in the reduction of GABA release. It is likely that the decrease in GABA release attenuates GABAB receptor mediated neurotransmission, thereby upregulating GABAB receptors. PMID- 12762221 TI - A case of malignant syndrome associated with a parkinsonism patient. AB - Malignant syndrome is usually triggered by an excessive use of neuroleptics and dehydration. In the patients with Parkinson's disease, it is sometimes provoked by discontinuation of anti-Parkinsonism agents. Nevertheless, there are few reports describing malignant syndrome in Parkinson's disease patients and no typical therapeutic strategy had been established in such cases. We recently experienced a case of malignant syndrome associated with a familial Parkinsonism patient, which was brought about by surgical invasion and a break in the administration of anti-Parkinsonism agents. Respiratory muscle paralysis, megacolon, and nocturnal delirium observed in this patient might have partially resulted from the use of dantrolene administered during the course of the treatment. Since an increase in the sensitivity to certain drugs is presumed in Parkinson's disease patients, we should be sufficiently cautious about drug administration. We should administer dantrolene, recognizing the fact that the use of dantrolene is one of the symptomatic treatments for muscle rigidity. PMID- 12762222 TI - [Post-transcriptional regulation by 3'-untranslated region of transcripts]. AB - Two different mechanisms have been well known to regulate the amounts of various transcripts in response to internal and external environmental stimuli. First, by binding of activated transcription factors to DNA regulatory regions including the cAMP response element, hormone response element, and activator protein-1 region upstream in various genes, the rate of transcription from DNA to mRNA is regulated. Secondly, the degradation of some mRNAs related to immune responses has been reported to be regulated by binding of RNA-binding proteins to adenylate uridylate-rich elements (AU-rich elements, AREs) located in the 3'-untranslated region (3'-UTR). The original study identifying the existence of a common regulatory nucleotide sequence in the 3'-UTR of inflammatory mediator transcripts pointed out that the AREs are characteristic of immune-related functional proteins. The number of transcripts containing AREs in the 3'-UTR has increased and several neuronal proteins including beta 2-adrenergic receptor, nerve growth factor, tyrosine hydroxylase, and nitric-oxide synthase II, have been reported to have AREs. We here reviewed the recent advances in the neuropsychopharmacological understanding of post-transcriptional regulation by RNA-binding protein and also pointed out the importance of this regulation in future studies using various stress paradigms. PMID- 12762223 TI - [Organization and evolution of the human immunoglobulin heavy-chain loci]. PMID- 12762224 TI - [Expression of foreign proteins in E. coli: through the expression of P450s]. PMID- 12762226 TI - [Excitatory amino acid transporters: development of regulatory molecules for their functional analysis]. PMID- 12762225 TI - [Host defence mechanisms orchestrated by the pulmonary collectins]. PMID- 12762227 TI - [Molecular studies on inborn errors of ketone body metabolism]. PMID- 12762228 TI - [Functional link between histone acetyltransferase and chromatin assembling factor in gene silencing]. PMID- 12762229 TI - [Substrate RNA recognition mechanisms and protein structures of SPOUT, a novel super-family of RNA methyltransferases]. PMID- 12762230 TI - [Extracellular processing protease PACE4: significance of diversity of SPC family proteases]. PMID- 12762232 TI - Pediatrics today in Bosnia and Herzegovina. Proceedings of the 2nd Congress. Sarajevo, November 12-14, 2002. PMID- 12762231 TI - [The integrated database for mutations in disease-causing genes: Mutation View/KMDB]. PMID- 12762233 TI - [Neural tube dysraphism: meningomyelocele and related disorders]. AB - Neural tube disorders develop as a result of failure of neural tube closure between 3rd and 5th gestational weeks. This failure can cause soft structure anomalies (spina bifida, lumbal meningocela) or possible can contain neural tissue (meningomyeloccla, encephaloccla). Etiology of this disorder is not clear enough, and probably has multifactorial roots. Besides genetic factors, there are impact of some nutritional causes like folic acid. 28 cases with neural tube dysraphism hospitalized during period August 1999 till August 2002. at the Pediatric Clinic KCU Sarajevo were analyzed through retrospective study. 19/28 (67.8%) of newborn were from controlled pregnancy but without folic acid supplementation, 4 of them (14.2%) had prenatal diagnosis. Dysraphic disorder was the most often accompanied by paraplegia 16/28 (57.1%), hydrocephalus 17/28 (60.7%), from which 6/17 (35.2%) with Arnold Chiary malformation. 13/28 (46.4%) had skeletal deformities. Active preoperative treatment was conducted in 20/28 (71.4%) cases, and the rest of them were treated with home palliative care because of parent's rejection of surgery or major accompanied anomalies presence. In order to decrease the incidence of dysraphic disorders it is necessary to conduct periconceptional folic acid prevention, and provide early prenatal diagnosis. Long term treatment of children with meningomyclocele requires multidisciplinary approach that includes surgeons, orthopedists, pediatricians, physical therapists, in order to improve life quality of survived children. PMID- 12762234 TI - [Detection and treatment of intracranial hemorrhage at the intensive care unit of the Pediatric Clinic in Sarajevo]. AB - Intracranial haemorrhage (ICH) is the common name for periventricular and intraventricular haemorrhage. We analyzed patients diagnosed as ICH in period January 2001 till May 2002. In 29/323 (8.9%) pts was verified ICH, 16/29 (55.1%) were male sex. Birth weight under 1000 grams had 6/29 (20.6%), birth weight 1000 1499 grams was 10/29 (34.4%), than 1500 to 2499 grams 8/29 (27.5%) and over 2500 grams 5/29 (17.2%). APGAR score were under 7 in 20/29 (68.9%), and four of tham 4/20 (20%) had severe and 16/20 (80%) pts had modest and mild forms of perinatal asphyxia. According to Papile classification of ICH, we found: I degree ICH had 12.29 (41.3%) pts, II degree 8/29 (27.5%) pts, while severe forms III and IV degree of of ICH had 9/29 (31.2%) pts. In 3/29 (10.4%) pts, posthemoragic hydrocephalus were registrated. Risk-factors for development of ICH were low birth weight, small gestational age and perinatal asphyxia. PMID- 12762235 TI - [Ultrasound differentiation between benign enlargement of the subarachnoid space and brain atrophy]. AB - Fluid collections surrounding the brain (pericerebral fluid collections) in infants can be caused by a variety of conditions: benign enlargement of the subarachnoid space, passive dilatation of the subarachnoid space due to brain atrophy, subdural hygroma and subdural effusion as a result of meningitis or subdural haematoma. An enlarged frontal subarachnoid space and normal or minimally enlarged ventricular size, with normal or increase head circumference is relatively common finding in infancy. Without associated brain anomalies, this finding predicts normal development of the child. In our study, we have followed up psychomotor development of 22 children, with increased diameters of subarachnoid spaces, detected by ultrasound examination of the brain, without associated brain anomalies. The inclusion criterion for study was enlarged subarachnoid space, measured at the conventional coronal section at the level of the interventricular foramen. The upper limits of each measurements were: 3 mm for sinocortical width, 4 mm for craniocortical width and 6 mm for the interhespheric width. The patients have been followed up to the age of 24 months. Each of them had normal development. 16 of them had normal head circumference. 6 of them had head circumference above the 97th percentile and their head circumference measurements have been plotted for the next 6 months after the diagnosis, to be certain that growth is paralleling the normal curve. It is important to differ benign enlargement of subarachnoid space from brain atrophy, which is quite "serious" diagnosis, with poor neurodevelopmental prognosis. The brain atrophy has been presented with passive dilatation of subarachnoid space and ventriculomegaly, as well. In the case of benign enlargement of subarachnoid space normal or minimally enlarged ventricular size is present. For proper interpretation of this ultrasound finding, correlation with head circumference is necessary. We recommend the head circumference percentile chart to be enclosed to the each brain sonography finding. The algorithm of the further neurological evaluation differs significantly in the case of benign enlargement of subarachnoid space and brain atrophy. In the case of benign enlargement of subarachnoid space further neuroimaging procedures are not needed (in our study it has been done for 6 patients). If the brain atrophy is suspected, further complete neurological examination is necessary. PMID- 12762236 TI - [Urinary tract infections and vesicoureteral reflux as a cause of renal parenchymal damage]. AB - GOAL: Infection of the urinary tract is a risk for the damage of parenchyma and kidney function. Dimercaptosuccinil acid (DMSA) scan was used based on the relation between vesicoureteral reflux (VUR), infection of the urinary tract (UTI) and parenchymal kidney damage. METHODOLOGY: The number of observed children was 79, 58 (73.41%) girls and 21 (26.58%) boys: following the first UTIs, recidiv infect and with positive VUR contrast voiding cistourethrography (VCUG) and ultrasound tests were compelted following the treatment of infect within 6 weeks and DMSA within 4 months. The classification of VUR was done of I to V degrees. The damage to the renal parenchyma after DMSA was defined as inhomogen distribution, reduced bonding with radio-farmake as well as portion of the kidney in the overall renal function smaller than 45%. RESULTS: Among 45.56% (36/79) patients VUR was registered. Reflux over III/V was registered among 27.84 (22/79) among whom 24.05% (19/79) were younger than 5 years of age. DMSA scan showed reduced kidney function bellow 45%, in one direction 33.33% (12/36) and 25.00% (9/36) in both directions. The correlation between clinically proved acute pielonephrities and damage to the parenchyma was not proven. It was proven in 30.37% (24/79) reoccurring urinal infection. CONCLUSION: DMSA is a sensitive method that should not be routinely used on every child with urinary infection. It should be left to assess the renal damage in cases of high degree reflux and reoccurring urinary infection. PMID- 12762237 TI - [Monitoring oxygen saturation in the pediatric population]. AB - Pulsoxymetry is noninvasive technique which is in use for percentage estimation of hemoglobin saturation of oxygen in arterial blood. It represents a therapeutical screening of treatment in paediatric population. The aim of this retrospective study is to evaluate the validity of noninvasive technique for determination of oxygen concentration in cardiac and pulomological patients. The study included 221 patients, with a mean age of 39 months (1.5-192) who were hospitalised during the period of 1.6.2000. till 31.12.2002. at Pulmoalergology and Cardiorheumatology department of Paediatric clinic CCU Sarajevo. Three groups of patients were evaluated: first one, which included 194/221 patients with obstructive and inflammatory lung diseases, 17/221 patients formed II group of pts with congenital heart anomalies (CHA) and the third one which had 10/221 patients with CHA associated with lung diseases. In all patients the diagnostic laboratory investigations were done including transcutaneous determination of oxygen saturation by pulsoxyimetry (PO) Nellcor and Johnson-Johnson, as well as capillary, with adequate treatment. The mean oxygen saturation (OS) in the first group of patients at admission by PO was 86%, capillary 73.6%, and after treatment OS PO was 91.6%. In the II group the mean OS at admission was 88.5% PO, capillary 83%, after treatment 94.9%. The mean OS at admission in third patient's group was 76%, and after treatment 91.2% which is statistically significant (F = 0.03) and is in concordance with modern therapeutical approach. Pulseoxymetry is noninvasive, painless, simple method which offers a valid continuous data needed for the adequate choice of treatment in paediatric patients. PMID- 12762238 TI - [Morbidity trends in the cardio-rheumatology department of the Pediatric Clinic in Sarajevo during the pre-war, war and post-war periods]. AB - In this work is presented the structure of the morbidity of the patients treated at the Cardiohematology department of the Pediatric Clinic in Sarajevo. It were followed four the most frequent groups of the diseases. The results show the trend of the increasing of the number of the hospitalized children with the congenital anomalies of the heart and juvenile chronic arthritis, with the decrease of the number of the ill become by streptococcal infections of the upper respiratory tract and the rheumatic illness. It is increased the number of the hospitalized children is the result of the beginning of the cardiosurgical way of the taking care of these patients. The decreased of the number of the ill become is the consequence of the improvement of the healthcare protection at the primary level with the continuity of the further trend of the decrease of the number of the patients ill become from acute rheumatic disease. PMID- 12762239 TI - [Allergy tests in children with seasonal allergic rhinitis]. AB - The aim of this study is to show positive allergy test od tree pollen, grass pollen and weed pollen in children with season allergic rhinitis. In the study is comprised 335 children, which are tested in Pulmoallergologic out-patient department of Paediatric clinic in Sarajevo in last three years from march 1999 to march 2002 year. In testing was used prick-test with Bencard firm allergen. Criterion for positive reaction was urtica, which appear 20 minutes after testing, with diameter equal or bigger than 3 mm, and erythema was 10 mm. Positive results on tree pollen was in 98 children (29.2%), on grass pollen in 210 children (62.6%), and on weed pollen in 27 children (8.0%). In group of tree pollens Coryllus avellana pollen (Hazel pollen) has the biggest number of positive results in 74 children (75.5%), in group of grass pollens Poa pratensis pollen (Mcadow grass pollen) has biggest number of positive results in 169 children (80.4%) and in group of weed pollens main allergen is Ambrosia clator (Rag weed) in 25 children (92.0%). In therapy of our children we preferred nasal topical corticosteroide Fluticason propionate Flixonase (GlaxoSmithkline) in one daily dose 50 mcg and we had fast and prolonged result in amelioration of symptoms. PMID- 12762240 TI - [Efficacy and tolerability of carbamazepine as the initial drug used in the treatment of epilepsy]. AB - Carbamazepine is the one of the most prescribed antiepileptic drugs in treatment of partial and generalized tonic-clonic seizures. Its efficacy in treatment of seizures was discovered incidentally during the trials of neuroleptic drugs. Generally it is well tolerated with relatively rare serious side effects. Therapy is introduced gradually, with mean dose between 10-20 mg/kg, and given this way it has the least side effects. Aim of the study was to assess interaction between efficacy, tolerability and overall efficacy of the first prescribed drug, in this case carbamazepine, in the group of patients with partial seizures and generalized tonic-clonic seizures. Patients hospitalized at University Pediatric Hospital in Sarajevo with newly diagnosed epilepsy and started therapy with carbamazepine in period from 07.1999 to 07.2002 were investigated. There were 89 patients that fulfilled the criteria. In 29 patients seizures remitted after the introduction of therapy (32.58%). After the subsequent correction of therapy further 8 patients were seizure free. In total, drug was efficient as a monotherapy in 68 patients (76.40%). Other antiepileptic drugs or polytherapy with carbamazepine were tried in the patients that were not seizure free. Adverse effects were reported in 28 patients (31.46%). The most frequent was benign leucopoenia (16 patients, 17.97%) that did not require changes in therapy. Rash was found in 8 patients (8.98%), out of which in 6 therapy had to be stopped. Compliance with therapeutic regime was slightly better with controlled release formulation (89.98%) then with simplex formulation (77.52%), but generally was good. We think that carbamazepine is still the drug of the first choice in treatment of patients with partial epileptic seizures and generalized tonic clonic seizures. It showed good overall efficacy with relatively rare serious side effects. PMID- 12762241 TI - [Growth hormone determination using the insulin-induced hypoglycemia test- results of a 3-year period]. AB - AIM: To present results of growth hormone (GH) stimulating test by insulin induced hypoglycaemia in the last three years. In the period from from 1992-1999. we could not perform any test for GH measuring because of lack of kits. PATIENT AND METHODS: Indications for measuring GH are body height < p3, bone age < p75 of proper for age or growth velocity < 4 cm/yearly. GH measuring test is performed by radioisotope method: results < 7 mlU/1 confirm absolute and 7-15 mIU/l partial GH deficiency. RESULTS: During three years' long period GH test was performed in 54 patients: 33 (61%) boys mean age of 11.8 years and in 21 girls (39%) mean age off 9.2 years. 32% of patients were from Canton of Sarajevo, 18.5% from Zenicki and the same percent from Unsko-sanski, 17% from Middle-bosnian Canton, 11% from Hercegovinian and 4% from Tuzlanski Canton. In 80% of patients body height was < p 3.80% of patient had delay in bone age more than 1 year. Results of test in 13 patients confirmed absolute and in 18 patients relative deficiency of GH. Only 4 patients with absolute GH deficiency are under GH therapy. CONCLUSION: Introducing GH measuring test again is useful. There is a great number of patients with absolute and relative GH deficiency (57%), so it is necessary to centralize evidention and medicine supply for those children. PMID- 12762242 TI - [Diagnostic value and cytomorphologic findings in bone marrow punctures in children with severe deficiency anemia]. AB - AIMS: To determine the diagnostic and prognostic value the bone marrow aspiration in children with severe deficiency anaemia. PATIENTS AND METHODS: In this retrospective study we analysed 75 children 0-4 years of age who were diagnosed and treated from severe anaemia (Hb < 8g/dl) at Haematologic outpatient and hospital department of Pediatric Clinic in Sarajevo in period of 01.01.1999. to 31.12.1999. RESULTS: We analyzed totally 75 children--37 (49.33%) boys and 38 (50.67%) girls. Morphologic finding of bone marrow aspirate smear showed: features of ferriprive flora in 24% of analyzed patients; dimorphic bone marrow features in 9.3% of patients; feriprivic flora with dyshematopoesis in 62.7% and megaloblasic anaemia features in 1.3% of analyzed patients. In two children we diagnosed leukemias from bone marrow aspirate; one was ALL/L2 and the second was AML/M2 leukemia. CONCLUSION: The most frequent found features of bone marrow aspirate were: feriprivic flora with dyshematopoesis. There was only one case of the megaloblastic bone marrow. But, the finding of dymorphic bone marrow indicated to pluricareny of ethiologic factors for beginning severe nutrititive anaemia. Special value of this paper was discovering of two cases of leukemia by the bone marrow aspirate analysis. PMID- 12762243 TI - [Infection as a complication of treatment of acute leukemia in children]. AB - AIM: Preventing and successful treatment of infections which are complications during treatment of acute leucosis in children. We analyzed a group of children who were treated at our Clinic in the last five-years period PATIENTS AND METHODS: In retrospective study we analyzed children who were treated at our Clinic during last five years. We analysed 56 children, and in 42 of them whole treatment was performed at our Clinic. 14 children were treated for some time in the other European Centers. RESULTS: We analyzed 56 patients. In 49 (89%) diagnose was ALL (Leucosis lymphatica acuta), and in 7 of them diagnose was AML (Leucosis myeloblastica acuta). The most frequent infections which were complications during treatment of leucosis: were infections of the mouth, infections of the respiratory tract-Pneumonia, sepsis and infections of urinary tract. The most frequent causes of infections were: Candida albicans- mouth, Staphylococcus epidermidis and Seratia marcenscens-sepsis, Escherichia colli urinary tract. CONCLUSION: According to our investigation we got conclusion is that infections are great problem in treatment of children with acute Leucemia. Prevention, supportive therapy and care are the most important in struggle with this series complication. PMID- 12762244 TI - [Occurrence of congenital anomalies in neonates in the Sarajevo region]. AB - Datas about frequency of congenital anomalies in the world are different because of the difference in ecological variations, socioeconomical differences, geographical differences and other conditions. Datas about incidence prevalence and mortality of the congenital anomalies during the infant period are insufficient in region Sarajevo. Goal of this study is to show frequency of congenital anomalies in region Sarajevo. Patients During the period from 01.01.1990-31.12.1999 in Sarajevo region was done the retrospective analysis of children who were born alive and dead, then the analysis of mortality of infants because of congenital anomalies, and children with anomalies. RESULTS: During the same period there were 47980 children born. There were 269 (5.6%) dead born children were 269 (5.6%). Incidence of mortality in infantil period was 120/10,000, and incidence of the children with congenital anomalies was 286/10,000 and was higher every year. There is no datas about mortality of congenital anomalies. CONCLUSION: This study shows that congenital anomalies in Sarajevo region are getting higher from year to year, and it is necessary to make a registry of congenital anomalies like these in the world. PMID- 12762245 TI - [The Patau syndrome]. AB - Known as D trisomy, Patau syndrome is the third chromosomopathy according to frequency. One of the 5000 newborn carries the trisomy 13. In over 80% cases there is fresh mutation with non separation in myeosis of older mother. The mosaic or translocation forms are not rare. The mail newborn with Patau syndrome is shown in this article. We notice: microcephalia, dolihocephalia, microphthalmia, cheilognatopalatoshisis, polydactilia, and found ultrasound changes at the brain, hearth and genitourinary system. Cytogenetic finding show: mail cariotype with aberrations 47, XY + 13, Sy Patau. PMID- 12762246 TI - [Neonatal hyperbilirubinemia: evaluation and treatment]. AB - We have analyzed 132 newborns with hyperbilirubinemia (BW < 2500 g, GW > 37 weeks) hospitalized in Neonatal Unit of Paediatric Hospital during 2001. In 78 out of 132 (59.1%) newborns, causal diagnosis were established. Rh isoimmunization caused hyperbilirubinaemia in 2.8% (3/132) cases, ABO isoimmunization in 15.95% (21/32), infections in 9.15% (12/32) and other (polycitaemia, kephalhaematoma, loss of weight > 8%, jaundice related to the brestafeeding) in 31.8% (42/132). Maximal serum bilirubin level was reached between fourth and seventh day of life. In 47/132 newborns (40.9%) non-specific hyperbilirubinaemia were diagnosed. All babies were successfully treated by fototherapy. Blood exchange transfusion was performed in only 2 cases. Rh isoimmunisation was not significant factor in etiology of hyperbilirubinemia because of global prevention of Rh negative mothers. Hyperbilirubinemia of "healthy full-term newborn" was most often seen in 40.9% cases. Diagnostic criteria for hyperbilirubinemia were revised, and fottherapy, as safe, non invasive method, replaced blood exchange transfusion, as an expansive procedure with low but constant mortality. PMID- 12762247 TI - [Secondary autoimmune hemolytic anemia caused by epidemic parotitis virus]. AB - We report a case of mumps virus (VM) caused secondary autoimmune hemolytic anemia (AIHA). A 12-year-old boy was hospitalized at Paediatric Clinic of Sarajevo because of haemolytic anaemia followed by disturbed consciousness. Idiopathic hemolytic anemia was diagnosed on the basis of anamnesis, detailed clinical examination and laboratory analysis. The boy responded well to the therapy (corticosteroides, immunoglobulines, transfusions of red blood cells without buffy coat) with improvement of general condition and clinical aspect. Ninth day since the beginning of illness, firstly one than another parotide gland were enlarged, painful, followed by swallow problem and erythema of Stensen's duct. Epidemic parotitis were diagnosed on the basis of high amylasa in serum and urine. CONCLUSION: AIHA could be seen as primary disease. But, in paediatric population it is more often only one manifestation of underlying disease which we should search for. PMID- 12762248 TI - [Epidemiology of childhood leukoses at the Pediatric Clinic in Sarajevo 1997 2001]. AB - AIMS: Present pictures of epidemiology by leukemia from children from 0-15 years old during last five years. PATIENTS AND METHODS: In retrospective studies analyzing children of leucosis. This information's we have taken from history of illness. 67 children are worked out which are diagnosed and cured of leukemia in Pediatric clinic in Sarajevo, during last five years. In a study are included most of children which are diagnosed and cured in our clinic like children's which are diagnosed by our clinic, but they are diagnosed by our clinic, but they are receiving parts of terraping in other country. RESULTS: Results are showing that the 67 children are analyzed, most often maligned illness' which are related with others malignancies, they are active in 44.3%, there are acute lymphatic leucosis they are active in 86.5% tested children. The biggest number of tested children are coming from Sarajevo and Tuzla around 25.3%. First phase of leukemia is approximately same in matter of years. In the city environment the incidents of illness is bigger. FINAL THOUGHT: The children's leukemia is always growing on our territory, which can be explained, growing number of children's in clinic, which are taking terrapin in our clinic and acute lymphoblast leukemia are dominated in clinic of analyze which present new imperative in the view pastern diagnostic and therapeutic improvement. PMID- 12762249 TI - [Incidence of early-onset neonatal sepsis caused by group B Streptococcus at the Pediatric Clinic of the University Clinical Center in Sarajevo]. AB - Group B Streptococcus (GBS) is the main ethiological agent of neonatal sepsis in the developed countries. Because of high mortality rate American College of Obstetrics and Gynaecologists and Centers for Disease Control published recommendation for prevention of neonatal GBS infection. Program recommends screening of all pregnant women for anogenital GBS infection including intrapartum treatment of pregnant women at high risk, with penicillin or ampicillin. Clindamicin or erythromicin may be used for women allergic to penicillin. Since there is no detailed informations of early-onset neonatal GBS infection in our country, we investigated its incidence in Pediatric Clinic in Sarajevo, from December 1999 (when first case of early-onset GBS infection in last decade was diagnosed) to August 2002. During that period 7 neonates with early-onset GBS infection were identified (incidence 0.52 per 1000 livebirths). Four of seven sick newborns died, 2/7 survived with severe neurological sequelae and only one female newborn survived without deficits. In the same period we registered 36 (2.4%) GBS colonised neonates out of 1530 admitted neonates. The number probably is higher because of discharge GBS colonized asymptomatic neonates from Maternity as a healthy to home. Results of this study intend the necessity of implementation guidelines for GBS prevention since early onset GBS infection is becoming as an increasing problems in our population. PMID- 12762250 TI - [The Stevens-Johnson syndrome--personal experience]. AB - Stevens-Johnson syndrome is a disorder of the skin and mucous membranes and a severe bullous forms include systemic toxic effects. In this study, we have analyzed our experiences according to ages, sex, incidence, ethological irritable factors, differential diagnosis, clinical treatment, in children with Stevens Johnson syndrome at the Pediatric Clinic in Tuzla. In the period January 1997-May 2002, we treated 14 patients with this syndrome. All patients were male, ages from 11 months to 11.6 years (average 4.5 years). As irritable factors were detected antiepileptics (Phenobarbitol and Tegretol) in 10 children (71.44%), antibiotics (Procain Penicillin and Cephalexin) in 3 children (21.42%), and reason was stayed unknown in 1 child (7.14%). The problem in differential diagnosis usually was toxic epidermal necrolisis (TEN) and graft versus host disease (GvHD). All patients demanded treatment in the pediatric intensive care unit supporting with specialists immunologists, ophthalmologists, dermatologists, infectologists. Treatment included, besides vital volume support, systemic administration of corticosteroids, antibiotics and intensive local therapy lesions of skin and mucous membranes, specially eyes in special sterile conditions. Outcome and results of our medical treatment of all patients were good. PMID- 12762252 TI - [Monitoring fluid balance in pediatric intensive care]. AB - The dynamic metabolism as far as bigger amount of water versus solid tissues in child demand much better skilfulness in calculating liquid input and excretion in intensive care. It is very important fact in prevention of life threatening conditions in children and adults, especially in conditions with multiorganic disorders, because their treatment is reciprocally opposite. Considering experiences of ICU UZ Gent-Belgium we made in Paediatric intensive care unit at Paediatric clinic in Tuzla unique method of monitoring liquid balance in children within specific conditions in which standard method of monitoring just the amount of planed liquid, body weight and diuresis is not reliable enough. With this new monitoring we prescribe dynamics of parenteral input of crystalloid and colloids as well as peroral income much more precise and much more confidence in positive result and prevention of harder disorders as well. In this paper we present our own method of observation as well as results in period July 1999. Till September 2002. We did the intensive volume control in 88 or 45 of total number of 2255 patients treated in PICU in this period. We followed the liquid balance 399 days or 9576 hours. Primary nephrologic diseases were indication for balance in 17 or 19.3% children, while in other 71 or 80.6% children, were other diseases like cardiac failure, diabetic ketoacidosis, chronic systemic diseases, the Toxic shock syndrome, the conditions of intracranial hypertension. We notice the signs of threatening renal failure in 7 or 7.9% of children and signs of manifesting renal failure in 10 or 11.3%. Our experiences tell that the model we made is applicable in working conditions of ICU. Evidence of organism responds the planned input is much better and safer than in standard way of observation. The possibility of in time intervention against threatening renal failure as well as other organic disorders is much greater. We recommended this model for all life threatening conditions in Paediatric intensive care units. PMID- 12762251 TI - [Association between chronic gastritis in childhood, Helicobacter pylori and ABO blood groups]. AB - Gastritis can be defined in the simplest way such as inflammation of gastric mucous membrane. There are numerous causes of gastritis and the most often is Helicobacter pylori infection. Studying of the blood groups also includes the question of their connection with certain diseases. Up to date it has been explained the role that blood groups have in haemolytic disease of newborn infant but for other diseases we know little, it is insufficient or we know nothing. The aim of this work was to analyze connection of chronic gastritis in childhood, Helicobacter pylori and ABO blood groups. It has been treated 68 patients out of which 57 (83%) were Helicobacter pylori positive (Hp+) and 11 (17%) were Helicobacter pylori negative (Hp-). Thirty patients had chronic gastritis out of which 22 (73%) were Hp+ and 8 (27%) Hp-. It prevailed blood group A and O, comparing with other blood group phenotypes connected with gastritis. PMID- 12762253 TI - [Pathologic ophthalmologic changes in prematurity]. AB - The personal author's experiences has been presented, with regards to the early discovering and adequate preventing of ophthalmology diseases at prematurely born children. Thanks to the development of perinatology, almost every organ can be monitored and functionally examined even before the child is born. Despite all problems, the work of ophthalmo-paediatritians is extremely challenging, because that is the only situation in which embryology of the eye can be seen "In Vitro" and in which the physiological development of the eye's function can been monitored. During the period from 1999 to 2002, it was examined 66 children in total, who had an anamnestical data about prematurity, as well as the data about delivery-weight. Out of these 66 children, there were 40 (60.6%) boys and 26 (39.4%) girls, with 1-4 of age. All children were examined by usual, in daily work available, examination methods. The biggest percentage of children (80%) were sent to us by the paediatritian-neonathologist, and 20% war sent from the Primary Health Care centers, or they were sent from other centers. At 52 (78.7%) of children, the certain changes on the eyes were found, while at 10 (15.1%) children no changes at all were found. At 4 (6.2%) cases, we found minor changes, but we monitored those children as well. Ophthalmology changes were found in almost all forms, from the most complicated (ROP, coloboma horioretinae, congenital glaucoma, congenital cataract), to the simpler ones on which are less difficult to treat (refraction changes, amblyopia, strabismus, ptosis etc). Since these changes are still present with prematuruses, it is necessary to intensively monitor this population, as a part of the multidisciplinary team, made of the experts of the different profiles (paediatritian-neonatologist, otologist, logopedist, ophthalmologist, etc), and which would be possible through the Register of Prematurity. Author introduces her own experience of ophthalmologic diseases in children who are categorized as "risky". A team of different specialists does treatment. PMID- 12762254 TI - [Psychosocial support and home-based physical therapy in children adolescents with childhood cerebral palsy in the Canton of Sarajevo]. AB - Problem of psychosocial support and home physical therapy of children and adolescent with Cerebral palsy we treated with Project with same name. Project is implementing together with Association of persons with Cerebral Palsy of Canton Sarajevo. Project is part of City of Sarajevo Project--"Sarajevo healthy town". 36 children and adolescents with Cerebral palsy, from 7 to 20 years, are included in six months home physical therapy. We included intellectually disabled and not disabled children and adolescents from Canton Sarajevo. Target population, except children, are parents and families of this children. Social worker and clinical psychologist are trainers of the Parents workshops. More psychosocial support we will have on Round table "Social integration of persons with Cerebral Palsy" and first brochure of the Association. PMID- 12762255 TI - [An original protocol for conservative treatment of fractures of the femoral diaphysis in children]. AB - Diaphyseal femoral fractures at children is very common, especially during the children play, traffic accidents and others. In spite on the dramatic clinical status (severe pain, deformity, pathological motions, and loss of stability) and great dislocation, conservative treatment almost always have good clinical result. Basis of this treatment is 3 week of indirect traction and after 6 weeks of plaster cast immobilization with permanent clinical and radiological evaluations. Operative treatment is reserved for cases of unsuccessfull conservative treatment and its complications. Resistutio ad integrum is almost always complete, thanks to good quality of this method, good biological potential of the child bone and good rehabilitation after immobilization. PMID- 12762256 TI - [The ALLHAT study: contributions, limitations and prospectives]. PMID- 12762257 TI - [Morphologic and functional abnormalities of the cardiovascular system in patients with hepatic cirrhosis]. AB - During liver cirrhosis many important changes occur in the cardiovascular system and these abnormalities appear more evident as portal hypertension and liver dysfunction progress. The cirrhotic heart develops a series of structural and functional abnormalities consisting in diastolic dysfunction and reduced myocardial reactivity during exercise, likely due to a diminished myocardial beta adrenergic receptor function. Nevertheless, the peculiarity of the cardiovascular involvement during liver cirrhosis is represented by a progressive development of a hyperdynamic circulation that seems to be due to portal hypertension rather than to liver insufficiency. In fact, it has been hypothesized that this syndrome raise from the venous portal bed and is primarily determined by an increase in blood volume that leads to an enhanced cardiac output. Later, as liver cirrhosis progresses, new important pathogenetic elements occur and lead to a reduction in peripheral vascular resistances and to the full clinical expression of hyperdynamic circulation. The pathogenesis of hyperdynamic circulation is very interesting for scientific research because of the complex and still in part unknown origin. In addition, this syndrome has an important clinical meaning for its severely adverse prognostic value and it represents the pathogenetic background for a number of severe complications of advanced liver cirrhosis. PMID- 12762258 TI - [Beta-blocker therapy and risk reduction in non-cardiac surgery]. AB - Cardiovascular complications are important causes of morbidity and mortality with major noncardiac procedures. Preoperative cardiac evaluation aims at assessing the patient's clinical risk profile in order to provide the more cost-effective strategy to reduce the risk of cardiac complications. Among different ways to reduce the incidence of perioperative cardiac complications, compelling evidence comes from the use of beta-blockers: in the absence of absolute contraindications, beta-blocker therapy should be administered to all patients at intermediate-high risk for coronary events who have to undergo noncardiac surgery. Even if the number of patients enrolled in these studies is relatively small, the use of beta-blockers before noncardiac surgery has been shown to be associated with a significant reduction in major cardiac events so that this therapy may reduce the need for additional noninvasive tests in some groups of patients. Coronary angiography and revascularization should have the same indications as if performed in the nonoperative setting. PMID- 12762259 TI - [Early prehospital thrombolysis in acute myocardial infarct: a moral obligation?]. AB - Acute myocardial infarction accounts for a large proportion of deaths from cardiovascular diseases. Occlusive thrombosis superimposed on a ruptured atheroma in an epicardial coronary artery is firmly established as the immediate cause of an acute myocardial infarction. Clinical research has focused on reducing the time to treatment, because necrosis of viable myocardial tissue mainly happens during the 30 to 90 min after coronary artery occlusion. Consequently, if the coronary artery can be reperfused during this period, extensive myocardial necrosis can be prevented and left ventricular function can be preserved. Indeed the mortality reduction by thrombolytic treatment compared with control is considerably higher in patients treated within 2 hours of symptom onset. Thrombolytic treatment during the first hour resulted in a 50% mortality reduction, which indicates 50 to 60 lives saved per 1000 patients treated. Early patency has crucial prognostic significance because the meta-analysis of all randomized trials of prehospital versus in-hospital thrombolysis shows that reducing treatment delay by 1 hour saves approximately 20 lives per 1000 patients treated. One way to reduce the delay is to bring the treatment to the patient in the prehospital setting. The safety and feasibility of prehospital thrombolysis strongly depend on the possibility of a rapid and correct diagnosis in the prehospital setting. To diagnose a myocardial infarction a standard 12-lead electrocardiogram is recorded and interpreted either on site by the emergency physician or, after telephone transmission, by a cardiologist on duty at the receiving coronary care unit. This approach has been proved to be safe. The most suitable prehospital thrombolytics are the third-generation agents given as a bolus, which have been tested in large hospital randomized control trials such as GUSTO-V and ASSENT III (reteplase and tenecteplase respectively), and the prehospital trial ASSENT III PLUS (tenecteplase). Hopefully future management of acute myocardial infarction with ST-segment elevation will include prehospital thrombolysis as a complementary part of any reperfusion strategy. PMID- 12762260 TI - [Health care quality in terminal cardiac decompensation: a "multidisciplinary" reflexion]. AB - Although improved, the prognosis of chronic heart failure is still malignant, the survival rate being similar to that of cancer. Due to the relevant epidemiology of this disease, the terminal care of heart failure has a significant impact, searching for the best quality of cure and care. PMID- 12762261 TI - [Usefulness and limitations of contrast echocardiography during dobutamine stress test]. AB - BACKGROUND: The aim of this study was to evaluate whether the use of contrast agent in addition to second harmonic imaging during dobutamine stress echocardiography can improve endocardial visualization and interobserver agreement in the evaluation of regional wall motion in patients with suboptimal or poor acoustic window. METHODS: Twenty-one patients with a poor or suboptimal acoustic window underwent dobutamine stress echocardiography. Echocardiographic images in parasternal long-axis and short-axis, apical 4-chamber and 2-chamber views were cine-looped at baseline and peak stress before and after injection of contrast medium (Levovist at a concentration of 400 mg/ml). Endocardial visualization and regional wall motion were evaluated by two blinded observers. RESULTS: The contrast medium improved segment visualization both at baseline (complete visualization in 74% of segments with contrast vs 71% without, p = NS) and at peak stress (76 vs 64%, p < 0.001). Contrast medium improved significantly segment visualization in apical 4-chamber view both at baseline (complete visualization in 87% of segments with contrast vs 72% without, p < 0.01) and at peak (89 vs 66%, p < 0.001) and in apical 2-chamber view both at baseline (81 vs 61%, p < 0.001) and at peak (89 vs 55%, p < 0.001). When individual segments were analyzed, endocardial visualization improved significantly in all segments of the anterior wall and in the mid and distal segments of the lateral wall both at baseline and at peak stress. The use of contrast medium did not improve significantly interobserver agreement in the evaluation of regional wall motion at peak stress (k = 0.63 vs 0.67 without and with contrast, respectively). CONCLUSIONS: The use of Levovist during dobutamine stress echocardiography improves significantly segment visualization in the apical views both at baseline and at peak stress and increases interobserver agreement in the evaluation of regional wall motion at peak stress. PMID- 12762262 TI - [Cerivastatin and the PRINCESS study: reflexions after 1 year]. PMID- 12762263 TI - [Infusion of flecainide in a patient with atrial fibrillation and latent Brugada's syndrome has determined modifications of the electrocardiogram similar to those of a septal myocardial infarct]. AB - ST-segment elevation in the presence of typical chest pain is a fundamental criterion for the diagnosis of acute myocardial infarction. We describe the case of a 62-year-old male with latent Brugada syndrome in whom the intravenous infusion of flecainide for paroxysmal atrial fibrillation caused ECG abnormalities similar to those of an acute septal myocardial infarction. The patient was not submitted to systemic thrombolysis since accurate echocardiographic evaluation was not suggestive of the presence of any wall motion abnormality and blood analysis did not reveal any increase in the levels of myocardial enzymes. The clinical outcome was not complicated and the patient was finally discharged. PMID- 12762264 TI - [High-output congestive cardiac decompensation caused by an atrio-venous fistula resulting from lumbar disk surgery]. AB - Although uncommon, arterio-venous fistula is a serious complication of lumbar disk surgery. Unless congestive heart failure develops, it often remains undetected. The present case report refers to a 30-year-old male who was referred to our division with a diagnosis of high-output congestive heart failure. It is important that surgeons, cardiologists and radiologists be aware of this potentially fatal complication. PMID- 12762265 TI - [Asymptomatic left atrial myxoma in the elderly]. PMID- 12762266 TI - [Practice guidelines for interpreting the electrocardiogram of the newborn infant]. PMID- 12762267 TI - Therapeutic strategies for patients hospitalized with worsening heart failure. PMID- 12762268 TI - Non-transplant surgery for heart failure and severe left ventricular dysfunction. Opportunities and limitations. AB - The need of alternatives to transplantation has lead to routinely reconsidering conventional surgery for selected patients with advanced heart failure. Although the recovery of ventricular contractility with revascularization sometimes restores ventricular function almost to normality while dramatically enhancing the quality of life, only a few patients with ischemic left ventricular dysfunction exhibit the amount of hibernating myocardium required for such a spectacular outcome taking place. Advanced remodeling is a further obstacle and is currently being addressed by means of operations designed to reduce ventricular volume and restore geometry. Hemodynamic measurements raise concerns, however, as to the diastolic function and the competence of the mitral valve late after reconstructive surgery. Finally, the repair of mitral valve regurgitation has produced interesting short-to-medium term results both in terms of survival and heart failure relief. Yet, their durability is uncertain, if not disappointing when the valvular disease has an ischemic etiology. Expectations of patients undergoing one of the aforementioned procedures, or a combination thereof, should not be averaged as they heavily depend on the specific patient profile under evaluation. PMID- 12762269 TI - Evidence-based evaluation of benefits in therapeutic interventions: methodologically controlled and non-randomly assigned reflections on the number needed to treat. AB - BACKGROUND: The number needed to treat (NNT), calculated as the reciprocal of the absolute risk reduction, is a parameter that provides quantitative information on the efficacy of therapeutic interventions. The introduction of this parameter was provoked by the demonstration that physicians were not too familiar with the percentage expressions of risk reduction, but preferred, in assessing the efficacy of a treatment in the context of a clinical trial, numbers that directly indicated patients. DESCRIPTION: Although the results of the evolution in the concept of the NNT have been available for various years, the diffusion of such parameters is still poor. The frequency of explicit reporting of the NNT in the published randomized controlled trials of five major biomedical journals has been recently assessed. This review has shown that in 359 eligible papers, the NNT was recorded in only eight articles. This represents a major drawback of biomedical literature, since the NNT allows health operators to understand how much effort is needed to prevent a given event. CONCLUSIONS: The NNT represents a measure of immediate and major clinical impact, and should always be reported, when appropriate, as an expression of the results of clinical trials. As a single number, it cannot stand alone as the sole justification for including a new treatment in the therapeutic armamentarium; still, it should be seen as an indicator by means of which the same treatment may be critically considered. It seems therefore appropriate to require that authors publishing results of clinical trials should provide readers with the information necessary to allow them to make responsible decisions; in this case, adopting the useful tool constituted by the NNT. PMID- 12762270 TI - Patients with chronic heart failure encountered in daily clinical practice are different from the "typical" patient enrolled in therapeutic trials. AB - BACKGROUND: The aim of this study was to compare the clinical characteristics of patients enrolled in randomized clinical trials on congestive heart failure treatment with those of real-world patients encountered in daily clinical practice. METHODS: We searched the Cochrane review methodology, Medline and SilverPlatter databases to obtain the clinical characteristics of both patients enrolled in therapeutic clinical trials and real-world patients with heart failure. We selected 27 clinical trials, and 8 prospective epidemiological studies or registries published between 1987 and 2001 which enrolled 53,859 and 18,207 patients, respectively. RESULTS: On average, compared to real-world heart failure patients, patients enrolled in clinical trials were younger (63 +/- 10 vs 75 +/- 11 years respectively, p < 0.0001), and more likely to be male (72 vs 54% respectively, p < 0.0001). Clinical trial patients showed a lower ejection fraction (26 +/- 7 vs 38 +/- 15% respectively, p < 0.0001) but a lower prevalence of NYHA functional class III-IV (62 vs 75% respectively, p < 0.0001) than real world patients. In clinical trial patients, the prevalence of ischemic heart disease (67 vs 42% respectively, p < 0.0001) and a history of previous myocardial infarction (62 vs 42% respectively, p < 0.0001) were higher than in real-world patients. Conversely, the prevalence of chronic atrial fibrillation (12 vs 31% respectively, p < 0.0001) and of diabetes (22 vs 24% respectively, p < 0.02) was lower in trial patients than in real-world patients. CONCLUSIONS: Our data suggest that most clinical trials on congestive heart failure, on which the guidelines for clinical practice are based, have generally included patients who are not representative of the whole spectrum of patients actually managed in clinical practice. PMID- 12762271 TI - Usefulness of exercise test in selected patients coming to the emergency department for acute chest pain. AB - BACKGROUND: The management of patients with acute chest pain is a common and difficult challenge for clinicians. In our emergency department (ED) a systematic protocol that involves the use of the exercise test for the management of patients with chest pain of suspected cardiac origin is presently running. The aim of the present study was to evaluate the feasibility of such a test in this setting, in terms of the safety and satisfactory follow-up of these patients discharged home. METHODS: Patients with chest pain lasting < or = 24 hours, aged > 18 years, without a history of trauma or of any other evident medical cause of chest pain and without high-risk characteristics were included in the present study. These patients, defined as low-risk patients for acute coronary events on admission, were evaluated in the ED area and submitted to serial ECG and blood sampling for the determination of the creatine kinase-MB mass and troponin I serum levels on admission and at 6 and 12 hours after admission. A symptom limited maximal exercise was performed in the patients with a negative clinical observation and typical chest pain or atypical chest pain but multiple coronary risk factors. RESULTS: In the year 2000, 1370 patients were evaluated in the ED for chest pain. In 150 (11%) an exercise test was performed. The test was positive in 24 patients (16%). The criteria for a positive test were only clinical in 3 patients, only ECG in 13 patients, and both in 8 patients. Inconclusive tests were observed in 27 patients (18%) and the test was negative in 99 patients (66%). There were no complications during the exercise test. At a median follow-up of 237 days (range 11-443 days), 11 clinical events were recorded (4 acute coronary syndromes and 7 revascularization procedures). Patients with a non-negative exercise test had a significantly shorter event-free survival (p < 0.005). CONCLUSIONS: The exercise test performed in selected patients coming to the ED with acute chest pain is safe and useful for further risk assessment. PMID- 12762272 TI - Diagnostic pathway of syncope and analysis of the impact of guidelines in a district general hospital. The ECSIT study (epidemiology and costs of syncope in Trento). AB - BACKGROUND: The ECSIT study was aimed at evaluating the hospital management of syncope patients, at comparing the appropriateness and costs of the hospital diagnostic pathway before (phase 1) and after (phase 2) the introduction of new guidelines and at analyzing the physicians' compliance to the guidelines. METHODS: All syncope patients admitted to the emergency room between August 1 and October 31, 1999 (phase 1) and between March 1 and May 31, 2000 (phase 2) were enrolled and their clinical records were analyzed in a blind fashion. RESULTS: During the study 538 consecutive patients came to the emergency room for syncope with a hospitalization rate of 53% in phase 1 (n = 151) and of 42% in phase 2 (n = 107). The in-hospital stay increased from 9 days in phase 1 to 11.3 days in phase 2 and diagnostic tests from 2.6 per patient (phase 1) to 2.9 per patient (phase 2) with total costs that rose from [symbol: see text] 3,474 to [symbol: see text] 3,647. Patients with no diagnosis decreased from 51 to 45.8% and the principal causes were identified as vascular brain disease (36.1 vs 33.7%) and neurally-mediated mechanisms (35.3 vs 42.2%). CONCLUSIONS: Despite the high costs of syncope management, the appropriateness and efficacy of the hospital diagnostic pathway remains far from ideal and simply introducing new guidelines seems unable to modify clinical practice. PMID- 12762274 TI - Beneficial effects of exercise beyond the pain threshold in intermittent claudication. AB - BACKGROUND: The quality of life and autonomy may be severely hampered in patients with intermittent claudication, but the amputation rate is very low. Supervised exercise training is effective, but still very rarely employed. Many authors think that in these patients exercise over the pain threshold may be dangerous. The aim of this study was to assess whether supervised, 3-month duration, 3 times/week, beyond the pain threshold exercise training is safe and whether it improves both the performance and quality of life in patients with claudication. METHODS: Forty-three patients with claudication, confirmed at Doppler study and/or angiography, have been evaluated by means of graded treadmill testing, the ankle-brachial pressure index at rest and after walking and a Walking Impairment Questionnaire before and after 3 months of treadmill training beyond the claudication threshold. RESULTS: Patients showed an 86% increase in time to onset of claudication pain (p < 0.00001), a 50% increase in total walking time (p < 0.000001) and improved questionnaire scores of pain intensity (%, p < 0.005), distance covered (+87%, p < 0.005), speed (+42%, p < 0.05), and stair climbing (+25%, p = NS). The basal and post-exercise ankle-brachial pressure index was not modified by training. Analysis of all subgroups of patients ( 65 years of age, with/without coronary artery disease and diabetes mellitus, pre-training time to onset of claudication pain 3 min, with angiographic/Doppler occlusion or stenosis) revealed a statistically significant increase in both time to onset of claudication pain and total walking time. CONCLUSIONS: Supervised physical training beyond the claudication threshold significantly improves the walking time and quality of life of patients with claudication. PMID- 12762273 TI - Exercise training counteracts the abnormal release of plasma endothelin-1 in normal subjects at risk for hypertension. AB - BACKGROUND: The hypothesis that in normotensive offspring of hypertensive parents exercise training could influence the systemic release of endothelin (ET)-1 during a provocative testing protocol was tested. METHODS: The provocative handgrip test was performed in four groups of healthy young age-matched males: offspring of hypertensive parents following a regular swimming exercise regimen (group A, n = 14); offspring of hypertensive parents and leading a sedentary lifestyle (group B, n = 11); normal volunteers with no family history of hypertension: sedentary (group C, n = 10), and following a regular swimming regimen (group D, n = 10). The plasma ET-1 was measured at baseline, after 4 min of handgrip exercise at 50% maximal capacity and following 2 (R2) and 10 (R10) min of recovery from handgrip. RESULTS: ET-1 plasma levels, within the normal range in all groups at baseline (group A 0.94 +/- 0.32 pg/ml, group B 0.84 +/- 0.26 pg/ml, group C 0.78 +/- 0.35 pg/ml, group D 0.85 +/- 0.26, p = NS) showed a progressive and significant increase in group B during and after handgrip exercise (peak handgrip 1.08 +/- 0.5 pg/ml, p = NS; R2 1.35 +/- 0.36 pg/ml, p < 0.05; R10 2.76 +/- 0.75 pg/ml, p < 0.01). Significant differences were found at R2 and R10 when the ET-1 levels measured in group B were compared to those observed in group A, group C and group D. Multivariate analysis demonstrated that the serum levels of ET-1 significantly contributed to predict handgrip-induced changes when the diastolic blood pressure was the dependent variable. CONCLUSIONS: Routine aerobic exercise appeared to counteract the handgrip-induced abnormal release of plasma ET-1 and may favorably affect the preclinical endothelial alterations seen in healthy offspring of hypertensive parents. PMID- 12762275 TI - Tricuspid valve annuloplasty using a partial flexible ring: mid-term follow-up. AB - BACKGROUND: Tricuspid valve annuloplasty (TVA) using a complete or partial rigid or flexible ring is becoming common practice in the surgical treatment of tricuspid insufficiency (TI). This study evaluates the immediate outcome and the mid-term clinical and echocardiographic follow-up after TVA performed using the Cosgrove-Edwards partial flexible ring. METHODS: From June 1998 to March 2002, 35 consecutive adult patients with TI > 2+ underwent TVA using this annuloplasty ring. TI was secondary to right ventricular dilation and/or dysfunction (functional TI) in 32 (91.4%) patients, and due to anatomic anomalies of the tricuspid valve (organic TI) in 3 (8.6%). Pulmonary hypertension was present in 34.4% of the patients with functional TI. Thirty-two (91.4%) patients were in preoperative NYHA functional class III or IV. The mean follow-up was 28.3 +/- 14.6 months. RESULTS: There were three (8.6%) in-hospital non-valve-related cardiac deaths, and one (3.1%) non-cardiac death during follow-up. The 3-year actuarial survival was 96.5%. TI was well controlled within grade 1+ in 30 (96.8%) survivors, but one presented residual grade 2+ TI. The NYHA functional class improved to 1.3 +/- 0.5 (p = 0.035). CONCLUSIONS: TVA performed using the Cosgrove-Edwards ring is a valid option for the surgical treatment of both functional and organic TI, even in case of pulmonary hypertension and at mid-term follow-up. PMID- 12762276 TI - Carotid plaque inflammation in a patient with unstable angina. AB - Coronary instability has been associated with multifocal plaque activation in the coronary circulation and in remote vascular districts, suggesting a systemic cause of instability, possibly inflammation. Magnetic resonance imaging offers a great potential for the detection of plaque inflammation. We describe the case of a 73-year-old female admitted for unstable angina, elevated levels of C-reactive protein and three-vessel disease, in whom carotid ultrasound examination revealed an atherosclerotic plaque of the left proximal internal carotid artery with an irregular profile and a heterogeneous echographic texture, determining a 50% stenosis. Magnetic resonance imaging of the plaque before and after contrast enhancement by gadolinium-DTPA showed the following signs of inflammation: an increased vessel wall thickness, an increased triple inversion recovery-fast spin echo signal intensity indicative of tissue edema, and a homogeneous plaque contrast enhancement indicative of an increased capillary permeability and neovasculature. As the carotid stenosis was < 70% and did not give rise to any symptom, the patient was submitted to coronary bypass surgery without concomitant carotid endarterectomy. Two days later she developed an ischemic stroke with right brachiocrural hemiplegia. In the present case report, the simultaneous presence of coronary instability and a carotid plaque with magnetic resonance features suggestive of inflammation, which was probably responsible for the stroke complicating cardiac surgery, may indicate a multifocal plaque instability. PMID- 12762277 TI - Inversion of the left atrial appendage following cardiac surgery. AB - Inversion of the left atrial appendage is a rare complication after open-heart surgery. To our knowledge only 16 cases, besides that reported herein, have been described so far. Echocardiographically, the inverted left atrial appendage appears as a mass mimicking a thrombus, a vegetation or a tumor of the left atrium. Lack of awareness of this entity can result in a misdiagnosis and unnecessary procedures. The case here reported deals with an inverted left atrial appendage occurring in an infant after repair of an atrial septal defect. PMID- 12762279 TI - Images in cardiovascular medicine. Life-threatening hemoptysis after the Fontan procedure. PMID- 12762278 TI - Valve adaptation as a cause of disc opening reduction in mechanical heart valves: the case of the Lillehei-Kaster valve. AB - At fluoroscopy a decreased disc motion in mechanical heart prostheses is often a sign of valve thrombosis. On occasion, however, despite an exhaustive diagnostic work-up, common causes of prosthetic valve thrombosis are not found. In these cases the valve disc abnormalities are thought to be due to functional changes. We here report our experience with 5 consecutive patients carrying the Lillehei Kaster prosthesis who had this fluoroscopic finding that was lately attributed to "valve physiologic adaptation". The time of onset, differential diagnosis and clinical/hemodynamic impact of valve adaptation are discussed. PMID- 12762280 TI - Massive coronary artery calcification in a young adult with end-stage renal disease undergoing hemodialysis. PMID- 12762281 TI - Electronic records. Protection on the road to patient safety. PMID- 12762282 TI - Clearing the hurdles to decision support. PMID- 12762284 TI - For some CIOs, thin is in. While thin client devices make inroads, the Web is redefining 'thin.'. PMID- 12762283 TI - NCVHS to extol a standard vocab. HHS committee completing its work culling alphabet soup of terms. PMID- 12762285 TI - Clinical guidelines gain mobility. PMID- 12762286 TI - Responding to a distant cry for help. PMID- 12762287 TI - HIV lipodystrophy etiology and pathogenesis. Body composition and metabolic alterations: etiology and pathogenesis. AB - The results of epidemiologic investigations have clearly indicated that the development of lipodystrophy is multifactorial. Factors related to HIV infection, hormonal influences, mitochondrial dysfunction, cytokine activation related to immune reconstitution, and individual genetic predisposition all have been hypothesized as etiologic. Recent studies suggest that immune dysregulation rather than HIV infection per se may be the predominant factor in the development of lipodystrophy. PMID- 12762288 TI - HIV lipodystrophy diagnosis and management. Body composition and metabolic alterations: diagnosis and management. AB - Body composition alterations are common in HIV infection and include AIDS wasting and lipodystrophy. Both are associated with metabolic alterations, including hypertriglyceridemia and reduced high-density lipoprotein cholesterol levels; insulin resistance and elevated low-density lipoprotein cholesterol levels are also associated with lipodystrophy. However, there is no accepted case definition for HIV-associated lipodystrophy, and patients may have one or all aspects at any given time. The inability of cross-sectional studies to capture the dynamic process of these alterations has hindered the search for a case definition. In the meantime, there are several approaches to treatment of the resulting abnormalities. Switching antiretrovirals has been most successful for improving metabolism, with little or not effect on fat distribution. Growth hormone treatment has successfully reduced visceral fat and buffalo humps but is not FDA approved for this indication. Metformin and rosiglitazone have produced some improvement in fat distribution as well as glucose metabolism. Other methods that have been tried with varying degrees of success (and little published data) include treatment with testosterone and its derivatives, weight reduction through diet and exercise, and plastic surgery. PMID- 12762289 TI - Coronary heart disease in HIV-infected persons. AB - Treatment of HIV-infected patients with HAART has prolonged survival, and clinicians must now be concerned with chronic diseases, some of which may be more prevalent in this population. This article focuses on coronary heart disease in persons with HIV infection. PMID- 12762290 TI - New concept in the pathogenesis of cardiovascular disease. AB - The advent of HAART has increased the lifespan of and enhanced the quality of life for patients with HIV infection. However, the gains achieved with HAART may be lost if all known risk factors for cardiovascular disease are not effectively managed in this population. This article reviews recent information on the contributions of insulin resistance, infection and inflammation, and psychiatric and psychological conditions associated with cardiovascular risk. It also summarizes available data on treatments of these risk factors. PMID- 12762291 TI - New concepts in the clinical aspects of cardiovascular disease. AB - Many patients with HIV infection, as well as those without this disease, may have coronary heart disease (CHD) or be at high risk for this condition. The high prevalence of and risk of CHD among patients with HIV infection are generally caused by traditional risk factors that can be readily managed with currently available therapies. New tools are also available for functional imaging and establishing diagnoses and prognoses in patients who do experience coronary events. Management of these persons requires careful attention to patient characteristics that may influence therapeutic outcomes. PMID- 12762292 TI - Infection control recommendations for patients with cystic fibrosis: Microbiology, important pathogens, and infection control practices to prevent patient-to-patient transmission. PMID- 12762293 TI - [Early pseudopuberty revealing juvenile granulosa tumor of the ovary]. PMID- 12762294 TI - [A new case of dirofilariasis in Corsica]. PMID- 12762296 TI - [Idiopathic Piringer-Kuchinka syndrome]. PMID- 12762295 TI - [Abuse of lactuca virosa]. PMID- 12762297 TI - Marine Biotechnology: Basics and Applications. Abstracts of an international symposium. Spain, February 25-March 1, 2003. PMID- 12762299 TI - Abstracts of the 18th Annual Meeting of the American Society of Hypertension. May 14-17, 2003, New York, New York, USA. PMID- 12762298 TI - Kansas nursing occupational outlook 1998-2008. PMID- 12762300 TI - Abstracts of the 34th Annual Meeting of the American Society for Neurochemistry. May 3-7, 2003, Newport Beach, California, USA. PMID- 12762301 TI - Institute of Medicine calls on government to lead charge toward quality and safety improvement. PMID- 12762302 TI - [Can Ageing be delayed? Abstracts of the 43rd Austrian Geriatric Congress. Bad Hofgastein, 22-26 March 2003]. PMID- 12762303 TI - Routine prostate screening may be unnecessary and harmful. PMID- 12762305 TI - Neuroprotection and Neurorepair. Abstracts of the 3rd Magdeburg Symposium. May 7 10, 2003. Magdeburg, Germany. PMID- 12762306 TI - Self-management improves functioning for age-related macular degeneration patients. PMID- 12762304 TI - Findings suggest revised atrial fibrillation approach. PMID- 12762307 TI - Abstracts of the 2nd Congress of the International Pediatric Transplant Association. Rio de Janiero, Brazil, 5-9 April 2003. PMID- 12762309 TI - Tunable diode laser spectroscopy 2001. Proceedings of the 3rd International Conference. Zermatt, Switzerland. July 8-12, 2001. PMID- 12762308 TI - A symposium: from prevention to management of chronic heart failure. Proceedings of the VII International Congress of Cardiologia 2000. Ostuni, Italy. May 31-June 1, 2001. PMID- 12762310 TI - Fabry disease: clinical heterogeneity and management challenges. Proceedings of the 2nd International Symposium on Lysosomal Storage Diseases. Cannes, France. April 2002. PMID- 12762312 TI - Abstracts of the 13th World Congress of the International Society of Brain Electromagnetic Topography, ISBET 2002. October, 2002. PMID- 12762311 TI - [Treatment at the departments of otorhinolaryngology and head and neck surgery and evidences: what constitutes an evidence?]. PMID- 12762315 TI - Minimally invasive endocrine surgery. November 18-22, 2002. Tokyo, Japan. PMID- 12762316 TI - [Drugs against urinary incontinence]. PMID- 12762314 TI - Failure to rescue. PMID- 12762317 TI - Proceedings of the 2nd C-Reactive Protein Hot Topic/Leiden Fibrinolysis Proteolysis Workshop. Leiden, The Netherlands. April 2002. PMID- 12762318 TI - Winning and losing magnet designation. PMID- 12762319 TI - Prevention of HIV: vaccine trials show only limited success. PMID- 12762320 TI - West Nile virus update: a new route of transmission is found. PMID- 12762321 TI - Abstracts of the 45th Annual Meeting of the Japanese Society for Neurochemistry. Sapporo, Japan. July 17-19, 2002. PMID- 12762322 TI - 12 ways you can prevent sports injuries. PMID- 12762323 TI - The Fernstrom Prize 2002 for Margareta and Birger Blomback. PMID- 12762324 TI - Heart-rate monitors. PMID- 12762326 TI - Blood-pressure monitors. AB - We tested 16 monitors and found that automatic arm models worked best; most wrist models lagged way behind. PMID- 12762328 TI - [Immune reconstitution in the era of highly active antiretroviral therapy]. PMID- 12762329 TI - [Helicoidal computed tomography in pulmonary arteriovenous malformation]. PMID- 12762330 TI - Delirium, depression often overlooked. PMID- 12762332 TI - [Progressive multifocal leukoencephalopathy caused by JC polyomavirus]. PMID- 12762331 TI - Idiopathic hepatic fibrosis with cholestasis in broiler chickens: immunohistochemistry of hepatic stellate cells. AB - Immunohistochemical examinations of hepatic stellate cells (HSCs) were performed on six enlarged livers from broiler chickens with malformation of the extrahepatic biliary tract (group 1) and on eight broiler livers affected with naturally occurring cholangiohepatitis without biliary malformation (group 2). The livers from both groups were grossly enlarged, firm and tan-coloured, and histologically revealed severe diffuse fibrosis with proliferation of bile ductules. HSCs positive for muscle actin and desmin actively proliferated in the perisinusoidal space and around newly formed bile ductules. There was no difference in the immunohistochemical reactivities and location of HSCs between the two groups. The findings suggest that the diffuse hepatic fibrosis found in group 2 as well as group 1 results from reactive proliferation of HSCs. PMID- 12762333 TI - "Double, double, toil and trouble': for modern practitioners of compounding, standards and training are critical [interview by Lisa Santandrea]. PMID- 12762334 TI - Opioid drugs in maintenance and detoxification treatment of opiate addiction; addition of buprenorphine and buprenorphine combination to list of approved opioid treatment medications. Interim final rule. AB - This interim final rule amends the Federal opioid treatment program regulations by adding buprenorphine and buprenorphine combination products to the list of approved opioid treatment medications that may be used in federally certified and registered opioid treatment programs. The Food and Drug Administration (FDA) recently approved Subutex[reg] (buprenorphine) and Suboxone[reg] (buprenorphine in fixed combination with naloxone) for the treatment of opiate dependence. These two products will join methadone and ORLAAM[reg] as medications that may be used in opioid treatment programs for the maintenance and detoxification treatment of opioid dependence. Opioid treatment programs that choose to use these new products in the treatment of opioid dependence will adhere to the same Federal treatment standards established for methadone and ORLAAM[reg]. The Secretary invites public comments on this action. PMID- 12762335 TI - Comment on the review "Anticoccidial vaccines for broiler chickens: pathways to success by R.B. Williams. (2002). Avian Pathology, 31, 317-53. PMID- 12762336 TI - Major conceptual change required to improve lung cancer: see a respiratory physician. PMID- 12762337 TI - In vivo assessment of lung inflammatory cell activity in patients with COPD and asthma. AB - The involvement of inflammatory cells in the pathogenesis of chronic obstructive pulmonary disease (COPD) and asthma is well established. This study aimed to quantify differences in inflammatory cell function in situ in these patients as compared to normal subjects. Positron emission tomography was used to assess neutrophil activity (18F-fluorodeoxyglucose (18FDG)) and macrophage accumulation (11C-PK11195) in six patients with COPD, six chronic asthmatics and five age matched normal control subjects. 18FDG uptake was greater in COPD than in normal subjects, with no increase in asthmatics. The mean slope of 18FDG uptake, corrected for volume of distribution, was 4.0 min(-1) in COPD patients compared with 1.5 min(-1) in control subjects and 1.7 min(-1) in asthmatics. Mean 11C PK11195 uptake (plateau tissue:plasma) was higher in four of six COPD patients (10.8) and three of five asthmatics (11.8) than the maximum value in control subjects (6.2). From this preliminary study the authors conclude that positron emission tomography may be useful to assess polymorphonuclear neutrophil and macrophage activity in vivo in chronic obstructive pulmonary disease and asthma, and may reveal differences in cell behaviour between the study groups. In addition, positron emission tomography may provide indices of disease activity for future therapeutic studies. PMID- 12762338 TI - Fluticasone reduces IL-6 and IL-8 production of cystic fibrosis bronchial epithelial cells via IKK-beta kinase pathway. AB - Inhaled fluticasone propionate (FP) is widely used to reduce pulmonary inflammation in chronic obstructive pulmonary disease, but the potential effects of FP on airway epithelial cells from patients with cystic fibrosis (CF) are unknown. In CF disease, a nonregulated inflammatory lung response occurs through exaggerated nuclear factor (NF)-kappaB activation and elevated pro-inflammatory cytokines production by airway epithelial cells. To determine whether FP reduces cytokine production in bronchial epithelial cells via NF-kappaB, the authors investigated the nonstimulated and the Pseudomonas aeruginosa lipopolysaccharide (LPS) stimulated production of NF-kappaB-dependent interleukin (IL)-6, IL-8 and RANTES (regulated on activation, T-cell expressed and secreted) along with the activation of NF-kappaB in non-CF and CF human bronchial gland epithelial cells. It was demonstrated that a relevant concentration of FP (10(-8) M) inhibited constitutive and P. aeruginosa LPS-induced IL-6 and IL-8 production of non-CF and CF bronchial epithelial cells. Interestingly, the expression of two IkappaB kinases (IKK)-alpha/beta, the degradation of cytosolic IkappaB-beta inhibitor and the NF-kappaB deoxyribonucleic acid binding activity were markedly reduced after FP treatment in both CF and non-CF bronchial epithelial cells. It was shown by the authors that fluticasone propionate exerts an anti-inflammatory effect by blocking a signal transduction leading to a reduced level of IkappaB-alpha/beta kinases in bronchial epithelial cells. In particular the strong effect on the IkappaB-beta kinase, which is known to be elevated in bronchial epithelial cells in cystic fibrosis patients, was observed. PMID- 12762339 TI - Induced sputum IL-8 gene expression, neutrophil influx and MMP-9 in allergic bronchopulmonary aspergillosis. AB - Allergic bronchopulmonary aspergillosis (ABPA) is a hypersensitivity reaction to the fungus Aspergillus fumigatus, causing severe asthma that may progress to bronchiectasis. Sputum neutrophilia can occur in association with sputum eosinophilia and correlates with the degree of bronchiectasis. The mechanisms of sputum neutrophilia in ABPA are not known. The aim of this study was to investigate the role of the chemokine interleukin (IL)-8 in sputum neutrophilia in ABPA. Induced sputum was obtained from subjects with ABPA (n=29), and compared to nonsensitised asthma (n=9) and healthy controls (n=21). Semiquantitative polymerase chain reaction was used to assess IL-8 gene expression in induced sputum and IL-8 protein was measured by enzyme-linked immunosorbent assay. Sputum IL-8 protein was significantly higher in ABPA compared to asthma and controls. IL 8 messenger ribonucleic acid/glyceraldehyde-3-phosphate dehydrogenase ratio was elevated in ABPA compared to asthma and controls. Sputum IL-8 correlated with sputum neutrophils, matrix metalloproteinase-9 levels and forced expiratory volume in one second. Interleukin-8 gene expression and protein release were increased in allergic bronchopulmonary aspergillosis and correlated with airway neutrophilia and airway obstruction. The interleukin-8-mediated neutrophil influx in allergic bronchopulmonary aspergillosis may induce lung damage via release of matrix metalloproteinase-9, potentially leading to bronchiectasis. PMID- 12762340 TI - Increased inflammatory markers in the exhaled breath condensate of cigarette smokers. AB - Cigarette smoking induces an inflammatory response in the airways that may play a key role in the pathogenesis of chronic obstructive pulmonary disease. Noninvasive markers of inflammation may, therefore, be useful in monitoring the airways of smokers as well as in the screening of subjects at high risk of developing airway obstruction. The aim of the present study was to determine whether the concentrations of the pro-inflammatory cytokine, interleukin (IL)-6, is increased in the exhaled breath condensate of smokers and whether the number of cigarettes smoked has any influence on the exhaled concentrations. The possibility that exhaled IL-6 levels are related to exhaled carbon monoxide (CO) and lung function has also been explored. Another inflammatory marker, leukotriene (LT), was also measured. Twenty-one smokers (39+/-7 yrs, 13 male) and 14 nonsmokers (45+/-6 yrs, eight male) were recruited. IL-6 and LTB4 levels in the breath condensate were measured with an immunoassay kit and exhaled CO examined by means of a modified electrochemical sensor. Higher IL-6 and exhaled CO concentrations were found in current smokers (5.6+/-1.4 pg x mL(-1) and 16.7+/ 5.5 parts per million (ppm)) than in nonsmokers (2.6+/-0.2 pg x mL(-1) and 2.1+/ 0.6 ppm). Elevated concentrations of LTB4 were also observed in smokers compared to nonsmokers (9.4+/-0.4 pg x mL(-1) versus 6.1+/-0.3 pg x mL(-1)). In addition, there was a correlation between IL-6 concentrations, the number of cigarettes smoked per day, exhaled CO, LTB4 and lung function. Exhaled interleukin-6 and leukotriene B4 levels may be useful noninvasive markers of airway inflammation in cigarette smokers. PMID- 12762341 TI - Exhaled NO and plasma cGMP increase after endotoxin infusion in healthy volunteers. AB - Nitric oxide (NO) is believed to be involved in the pathophysiology of sepsis. This study evaluated the activity of the NO pathway in a human endotoxin model. At baseline and after endotoxin, on-line measurements of exhaled NO (eNO) were made using a chemiluminescence technique with a single-breath method. NO-free air was inhaled prior to exhalation against a resistance. NO in orally and nasally exhaled air and in rectal gas was investigated. Plasma nitrite, nitrate, and guanosine 3', 5'-monophosphate (cGMP) and the events after diclophenac administration were also studied. Endotoxin infusion resulted in tachycardia and fever. An early increase in oral eNO concentration was observed and oral eNO decreased after diclophenac administration. NO exhaled nasally, NO in rectum gas and nitrite/nitrate levels remained unchanged over the study period, cGMP increased after 4 h. These findings suggest an early increase in nitric oxide production from the lungs, probably due to increased activity of the constitutive nitric oxide synthase upon endotoxin stimulation. In contrast, nitric oxide production in the upper airways, measured as nasally exhaled nitric oxide and nitric oxide in rectal gas, remained unchanged. Further studies will elucidate if exhaled nitric oxide is a valuable marker of sepsis-induced lung injury and if monitoring of treatment is possible. PMID- 12762342 TI - Biological markers in nasal secretions. AB - Biological markers in nasal secretions provide valuable information on nasal pathophysiology. However, published data on biomarker concentrations in nasal fluids are remarkably inconsistent, and the bias due to different sampling techniques, has not yet been systematically evaluated. Concentrations of various protein were repeatedly determined in nasal secretions of 16 healthy volunteers. The proteins were detected by using: 1) alpha2-macroglobulin as a marker for plasma contamination; 2) lactoferrin as a marker for glandular secretion; 3) lactate dehydrogenase as a marker for tissue injury; and 4) interleukin (IL) 1beta, IL-8, tumour necrosis factor-alpha, and eosinophil cationic protein and tryptase as indicators for tissue inflammation. A total of four different sampling methods, including nasal lavage (NL) and a new polyurethane foam sampler technique (PFST) were employed. Analyte concentrations in NL were approximately 10-times lower than in specimens obtained by PFST. Due to the unpredictable dilution during NL, various analytes were below the detection limit of the high sensitivity assays employed. With PFST, concentrations below the detection limit rarely occurred. The specimens did not significantly differ regarding plasma contamination, glandular secretion or tissue injury. The considerable variability of reported analyte concentrations in nasal secretions mainly results from different sampling techniques. To collect nasal secretions, samplers are considered superior to nasal lavage techniques. PMID- 12762343 TI - Lung cancer: the importance of seeing a respiratory physician. AB - Patients with lung cancer present to and are managed by a variety of clinicians. In this study the effect of involvement by a respiratory physician on the diagnosis, staging, treatment and survival of a large unselected group of lung cancer patients was investigated. The study population was derived from the Scottish Cancer Registry. A total of 3,855 patients diagnosed during 1995 with lung cancer were studied. The data were validated and supplemented by references to medical records. The study found that a respiratory physician had been involved in the initial management of 2,901 (75.3%) patients. These patients were found more likely to have had the cancer diagnosis confirmed by histological methods and to have received active treatment with surgery, radiotherapy or chemotherapy. Survival, 1 yr after diagnosis was higher in patients who saw a respiratory physician (24.4 versus 11.1%) and benefit was found to have remained 3 yrs after diagnosis (8.1 versus 3.7%). Although the patients who had not seen a respiratory physician were generally older, and had more extensive disease, after correcting for age, stage and other prognostic factors, the relative hazard ratio of death for those not managed by a respiratory physician was 1.44. The data from this study supports the recommendations of recent lung cancer guidelines for the early involvement by a respiratory physician. PMID- 12762344 TI - Epidermal growth factor receptor expression in pre-invasive and early invasive bronchial lesions. AB - The 1999 World Health Organization/International Association for the Study of Lung Cancer histological classification of preneoplastic bronchial lesions has been shown to be reproducible but little is known about its biological significance. The current study evaluated the correspondence between the morphological changes of the bronchial epithelium and epidermal growth factor receptor (EGF-R) expression. Thirteen normal bronchial epithelia, 19 hyperplasia, 16 metaplasia, 10 mild dysplasia, one moderate dysplasia, 10 severe dysplasia (SD), 14 carcinoma in situ (CIS) and 11 microinvasive tumours were assessed. A global EGF-R score obtained by the sum of the positivity score plus the EGF-R staining intensity score was calculated for each lesion. A global EGF-R score of >5 was reached only in one metaplasia, in six SD, in six CIS and in six microinvasive tumours. There was no difference in EGF-R expression between normal, hyperplastic and metaplastic epithelia versus mild dysplasia or between severe dysplasia versus CIS and microinvasive tumours but there was a statistically significant difference between mild versus severe dysplasia. This study demonstrates that epidermal growth factor receptor expression rate changes with the stage of the bronchial lesion, increasing from normal epithelium to carcinoma in situ and microinvasive tumours with a statistically significant difference between mild versus severe dysplasia. PMID- 12762345 TI - Plasma TARC concentration may be a useful marker for asthmatic exacerbation in children. AB - Recent studies suggested the T-helper cells type-2 lymphocytes-specific thymus and activation-regulated chemokine (TARC) and monocyte-derived chemokine (MDC) are useful inflammatory markers for chronic asthma. However, their roles in assessing the severity of acute asthma are unknown. This study aims to evaluate the serial changes of plasma TARC and MDC concentrations in children with asthmatic exacerbation. All patients with acute asthma were treated with systemic corticosteroid for 5 days. The severity of asthmatic exacerbation was classified according to the Global Initiative for Asthma guidelines. Plasma TARC and MDC concentrations were measured by sandwich enzyme immunoassays. Sixteen children, with a median (interquartile range) age of 9.3 (7.2-10.6) yrs and asthmatic exacerbation, were recruited. Plasma TARC concentration showed inverse correlation with peak expiratory flow rate at presentation. The median plasma TARC concentration was highest during the acute attacks (46 pg x mL(-1)) as compared to those levels at 1 (31 pg x mL(-1)) and 5 weeks (32 pg x mL(-1)) following treatment. The median plasma MDC level similarly decreased from 698 pg x mL(-1) at baseline to 261 pg x mL(-1) 1 week later, but increased back to 574 pg x mL(-1) at 5 weeks. These results suggest that plasma T-helper cells type-2 lymphocytes-specific thymus and activation-regulated chemokine but not monocyte derived chemokine concentration may be a useful inflammatory marker in assessing asthmatic exacerbation in children. PMID- 12762346 TI - Wheeze monitoring in children for assessment of nocturnal asthma and response to therapy. AB - The utilisation of nocturnal wheeze monitoring and quantification for assessment of asthma activity was studied in symptomatic school-aged children before and during treatment. Twelve children 6-14 yrs of age with mild or moderate untreated asthma were studied at home three times: before, 48 h and 6 weeks into treatment with 5 mg montelukast daily. Lung sounds were recorded overnight by an automatic wheeze detection device (PulmoTrack). Per cent wheezing within each respiratory cycle was calculated every 30 s throughout the night and a Nocturnal Wheeze Index (NWI) was calculated for the total night. The results were compared with spirometric indices (forced expiratory volume in one second (FEV1), forced vital capacity), bronchial reactivity (provocative concentration causing a 20% fall in FEV1 by adenosine 5'-monophosphate (PC20)) and daily symptom scores, performed in parallel at each stage of the study. The pretreatment NWI was 814+/-898 (mean+/ SD), which declined to 318+/-199 2 days after onset, and to 137+/-101 after 6 weeks of treatment. The NWI in seven healthy children was 47+/-43. The FEV1, PC20 and symptom scores improved in parallel. Wheeze monitoring provides quantitative and noninvasive information about the extent of nocturnal wheezing in children, correlates well with conventional indices of asthma activity and can assist in assessing efficacy of treatment. PMID- 12762347 TI - Effect of intravenous hydrocortisone on nocturnal airflow limitation in childhood asthma. AB - Low endogenous cortisol levels appear to contribute to the pathophysiology of nocturnal asthma. Lower cortisol levels are associated with lower forced expiratory volume in one second (FEV1) levels in children with asthma. The aim of the present study was to identify whether substitution of low serum cortisol with intravenous hydrocortisone decreases 24-h FEV1 variation and/or indirect measures of airway inflammation. Hydrocortisone was given over 24 h in a double-blind randomised crossover design to 26 subjects. FEV1 was measured every 4 h during 24 h; blood eosinophils and airway responsiveness to methacholine and adenosine 5' monophosphate (AMP) were measured at 04:00 h and 16:00 h. Cortisol levels increased during the night and morning hours. FEV1 values were higher at all time points in children with nocturnal asthma (n=10; 24-h FEV1 variation > or = 15%) which was significant at 08:00 h, unlike in the non-nocturnal asthma (NA-) group (n=16). Numbers of eosinophils (10(9) x L(-1)) at 04:00 h decreased in the asthma group (median 0.61 (range 0.05-1.42) versus 0.52 (0.05-1.79)). Provocative dose causing a 20% fall in FEV1 (PD20) methacholine did not change, whereas PD20 AMP improved only at 16:00 h in the NA- group (72.0 (0.13-144.0) versus 144.0 (2.25 144.0) mg x mL(-1)). These results show that substitution of lower endogenous 24 h values of cortisol contribute to higher forced expiratory volume in one second values and a decrease of blood eosinophils as an inflammatory marker in more severe airway obstruction. PMID- 12762348 TI - Adrenal suppression from high-dose inhaled fluticasone propionate in children with asthma. AB - This cross-sectional study was designed to examine the prevalence of adrenocortical suppression in children with asthma treated with high-dose inhaled fluticasone propionate (FP). Children and adolescents (n=50) with asthma, treated with inhaled FP at a dose of > or = 1,000 mg a day for > or = 6 months, were enrolled. Early morning serum cortisol was performed. Subjects with a serum cortisol of < 400 nmol x L(-1) had a tetracosactrin stimulation test. Fifty subjects of mean age 13.1 yrs were treated with a mean dose of 924.7 microg x m( 2) x day(-1) FP for a mean duration of 2 yrs. Of the 50 subjects, 36 (72%) had serum cortisol levels of < 400 nmol x L(-1) and underwent tetracosactrin stimulation test. Of these, 6 (17%) demonstrated a less than two-fold increase in serum cortisol from baseline and peak cortisol level of < or = 550 nmol x L(-1) at 30 or 60 min poststimulation. There was a significant negative correlation between the dose of FP x m(-2) and stimulated peak cortisol level. Biochemical evidence of adrenocortical insufficiency was demonstrated in 12% of the subjects, indicating that high-dose fluticasone propionate use may be associated with dose dependent adrenocortical suppression. PMID- 12762349 TI - Long-term follow-up of toluene diisocyanate-induced asthma. AB - Eighty-seven cases of occupational asthma induced by toluene diisocyanate (TDI) were diagnosed by an inhalation challenge with TDI and methacholine. After an average follow-up interval of 11 yrs, all subjects were re-examined. Of the 87 subjects examined, 13 (15%) had remained in the same job, 44 (50.5%) had been removed from exposure for <10 yrs and 30 (34.5%) had been removed for >10 yrs. The proportion of subjects who experienced symptoms of asthma and those who were hyperresponsive to methacholine was significantly lower. Of the patients, 59% used short-acting bronchodilators, 8% long-acting bronchodilators and 18% were on regular inhaled glucocorticoids. Thus, multiple regression analysis showed a positive correlation between forced vital capacity (FVC) and forced expiratory volume in one second (FEV1) at follow-up and FVC and FEV1 at diagnosis, and a negative correlation with smoking and with therapy with bronchodilators. Stepwise logistic regression showed that the follow-up provocative dose causing a 20% fall in the FEV1 (PD20) could be predicted from baseline PD20. These results indicate that respiratory symptoms and airway hyperresponsiveness to methacholine persist in subjects removed from exposure to TDI for >10 yrs. A more favourable prognosis was associated with a better lung function and a lower degree of airway hyperresponsiveness to methacholine at diagnosis. PMID- 12762350 TI - Airway inflammation in waste handlers exposed to bioaerosols assessed by induced sputum. AB - Work-associated lower airway inflammation in waste collectors was examined by induced sputum and correlated with the bioaerosol exposure. Organic waste collectors (n=25) underwent induced sputum collection and spirometry before work on Monday and the following Thursday. Total cells, cell differentials, interleukin (IL)-8 and eosinophilic cationic protein were determined. Personal full-shift exposure measurements were performed Monday, Tuesday and Wednesday and analysed for total bacteria, fungal spores, endotoxins and beta(1-3)-glucans. The percentage of neutrophils (46-58%) and the IL-8 concentration (1.1-1.4 ng x mL( 1)) increased from Monday to Thursday. Forced expiratory volume in one second (FEV1) was significantly reduced on Thursday, and the decrease in FEV1/forced vital capacity correlated with the increase in the percentage of neutrophils. The median exposure to endotoxin (range 7-180 EU x m(-3)) and beta(1-3)-glucan (range 5-220 ng x m(-3)) was correlated with the increase in IL-8. Bioaerosol exposure during waste collection induced an inflammatory response in the lower airways, characterised by neutrophils and interleukin-8 secretion, that influenced the lung function. The inflammatory response was related to microbial components in the bioaerosol and was more pronounced for endotoxin than beta(1-3)-glucan exposure. No associations were found for mould spores or bacteria. PMID- 12762352 TI - Chlorine inhalation produces nasal congestion in allergic rhinitics without mast cell degranulation. AB - Seasonal allergic rhinitic (SAR) subjects are more sensitive to nasal irritants than nonrhinitic (NR) subjects; however, the mechanism underlying this difference is unclear. This study sought to determine whether irritant-induced nasal congestion involves mast cell degranulation. Eight SAR and eight NR subjects were exposed to both 1.0 parts per million chlorine and filtered air in separate visits; exposures were via nasal mask and lasted 15 min. Rhinomanometry was performed before, immediately after and 15 min after exposure. Following > or = 2 weeks, exposures and symptom reporting were repeated with nasal lavage, rather than rhinomanometry, pre- and postexposure. A separate substudy using rye grass antigen provided a positive control. Mast cell tryptase was measured in nasal lavage fluid from both substudies using an automated fluoroenzyme immunoassay. Chlorine provocation significantly increased nasal airway resistance in SAR but not NR subjects. Conversely, tryptase levels in nasal lavage fluid were unaffected. Nasal allergen challenge significantly increased both nasal obstruction and nasal lavage tryptase in SAR subjects. Irritant-induced nasal congestion is more pronounced among seasonal allergic rhinitic than nonrhinitic subjects. However, unlike nasal allergen challenge, the mechanism of response to chlorine does not appear to involve mast cell degranulation. PMID- 12762351 TI - Lung accumulations of eosinophil granulocytes after exposure to cornstarch glove powder. AB - Starch is a main component of wheat flour, which, besides being an occupational allergen can also induce irritative symptoms in the airways. A purified starch product (cornstarch glove powder) was used to investigate whether starch alone could induce airway inflammation. The aim of the study was to investigate a role for starch in wheat flour-induced airway inflammation. Ten healthy individuals were exposed to cornstarch glove powder in a whole-body exposure chamber. Bronchoscopy with bronchoalveolar lavage (BAL) was performed 2-3 weeks before and 1 day after exposure, and the BAL cells were counted differentially. In addition, the expression of activation, adhesion and subset markers on alveolar macrophages and BAL T-cells were investigated using flow cytometry. A three-fold increase in BAL cell concentrations was found, with a selective accumulation and activation of eosinophilic granulocytes, as well as an influx of nonactivated monocytes and polyclonal CD4+ T-cells into the airways. The results show that inhalation of cornstarch glove powder leads to the development of a subclinical inflammation in the airways, with an accumulation of eosinophilic granulocytes. The authors suggest that such exposure may be an interesting model for studying factors contributing to lung accumulations of eosinophil granulocytes in humans. PMID- 12762353 TI - Relationship between FEV1 reduction and respiratory symptoms in the general population. AB - Obstructive airways diseases typically present with dyspnoea, cough and wheeze, and are defined by a reduced forced expiratory volume in one second (FEV1/forced vital capacity (FVC) ratio. Traditionally, however, the severity of chronic obstructive pulmonary disease is graded by the FEV1 % predicted rather than the FEV1/FVC ratio, whilst other potentially valid measures of FEV1 impairment, such as the absolute difference of FEV1 from predicted or the absolute level of FEV1, tend not to be used. The authors have therefore explored the relationship between these different measures of FEV1 impairment and chronic respiratory symptoms in a general population sample. FEV1 and FVC were measured and questionnaire data were obtained on cough, wheeze, shortness of breath and general self-reported breathing trouble in a cross-sectional survey of 2,633 adults aged 18-70 yrs from a district of Nottingham, UK. Odds ratios for each symptom were calculated for declining levels of absolute FEV1, FEV1 % pred, absolute difference of FEV1 from predicted, and FEV1/FVC ratio. They were plotted to display the shape and strength of these relationships before and after adjustment for each other measure. The odds of symptoms increased with declining levels of all FEV1 measures, particularly for wheeze and general breathing trouble. Although this study was not sufficiently powerful to detect significant differences between measures, these relationships were consistently strongest, before and after adjustment, for FEV1 % pred, particularly below a threshold of approximately 75%. The authors conclude that forced expiratory volume in one second % predicted appears to be the measure of airflow impairment most closely associated with chronic respiratory symptoms in the general population. PMID- 12762354 TI - Nutrition and respiratory health in adults: findings from the health survey for Scotland. AB - There is a growing body of evidence to support the hypothesised links between consumption of antioxidant rich foods and the occurrence of obstructive airway disease. The main research question was to examine the relationships between two types of dietary exposure and two indicators of respiratory morbidity in Scottish adults. The relationships between dietary consumption of fruit, vegetables and fish, and plasma levels of vitamins A, C, E and beta-carotene, and pulmonary function (forced expiratory volume in one second (FEV1)) and symptoms (phlegm production and shortness of breath with wheezing), were examined in a random population sample of adults. A dose/response relationship was found between fruit consumption and pulmonary function. In comparison with eating fruit rarely or never, eating fruit at least once per day, 1-6 times per week, and 1-3 times per month were associated with differences of 132, 100 and 63 mL in FEV1, after adjustment for known confounders and dietary intake of vegetables and fish (n=6186). An SD score change in plasma vitamin C was associated with a 49 mL difference in FEV1 (n=930). Fruit and vitamin E were associated with a reduced prevalence of phlegm production for 3 months or more per year. The most beneficial combination of dietary components may be found in natural foodstuffs, particularly fresh fruit. PMID- 12762356 TI - Replacement of physiologically autoinhaled nitric oxide in intubated patients. AB - Autoinhaled nitric oxide (NO) is produced mainly in the upper airways. Orotracheal intubation disrupts the natural autoinhalation of NO from the naso- and oropharynx. The effect of disrupting and then replacing autoinhaled NO on arterial oxygenation was investigated in intubated patients. Two groups of nine patients without lung disease were examined during anaesthesia using an inspired oxygen fraction of 0.50. In both groups, the individually produced NO of the whole respiratory tract and the upper airways was determined. The amount of NO normally autoinhaled from the upper airways was replaced for 5 min after orotracheal intubation in one group. The amount of NO from the upper respiratory tract was 47+/-19 parts per billion (ppb) in the test group and the replaced NO concentration was 48+/-20 ppb. No significant increase in arterial oxygen tension could be detected during the replacement of the autoinhaled NO. Haemodynamic parameters remained unchanged. In the control group, the NO from the upper airways was 34+/-16 ppb. In contrast to the test group, it was not replaced after intubation. These findings suggest that in healthy subjects the autoinhalation of nitric oxide does not play an important role in arterial oxygenation during anaesthesia. PMID- 12762355 TI - Passive smoking and respiratory symptoms in the FinEsS Study. AB - The aim of the present study was to examine the relationship between reported environmental tobacco smoke (ETS) exposure and respiratory symptoms. In 1996, a postal questionnaire was randomly distributed in three areas of Estonia to a population-based sample, of which 4,995 females and 1,822 males had never smoked. The main outcome measures were current respiratory symptoms and the amount of reported ETS exposure outside the home. ETS exposure at home was more common in females (31% versus 19%), while exposure outside of the home was more common in males (53% versus 7%). Females reported more symptoms from tobacco smoke than males (37.7% versus 21.6%). If ETS exposure outside of the home exceeded 5 h daily, the risk for wheeze (odds ratio (OR) 2.67, 95% confidence interval (CI) 1.98-3.61) and physician-diagnosed asthma (OR 1.79, 1.02-3.16) were increased. ETS exposure outside of the home was shown to be strongly related to almost all respiratory symptoms in a dose/response manner. ETS exposure at home did not show significantly elevated ORs for any respiratory symptoms. This study shows that females seem to be more troubled by environmental smoke exposure than males and provides further evidence of the serious health hazards associated with environmental smoke exposure. Indeed, the findings of this study support a ban on smoking in the workplace and public areas. PMID- 12762357 TI - Determinants of altered quality of life in patients with sleep-related breathing disorders. AB - Recent reports have suggested that altered quality of life and well-being are reported by patients with sleep-related breathing disorders (SRBD). There seems to be no data available in the literature on factors underlying these behavioural consequences. In this study, health-related quality of life (HRQL) scores were examined in SRBD patients in order to establish which factors are implicated in these disturbances. The study group consisted of 130 patients: 49 snorers and 81 patients with obstructive sleep apnoea. The Medical Outcome Survey Short Form-36 questionnaire was administered the morning after the sleep study and scores for the eight dimension scores were obtained. Patient's data were compared to normative sex- and age-matched data. In comparison with normal values, scores for all HRQL dimensions were decreased in SRBD patients, with a greater impact on subscores for "vitality", "physical role", "social functioning", "mental health" and "role emotional" dimensions. While impairment in physical function was mostly influenced by sleep stage and obesity, subjective daytime sleepiness mainly affected the other dimensions. The authors conclude that the altered health related quality of life of sleep-related breathing disorder patients is a multifactorial phenomenon depending on the interaction of sleep stages, daytime sleepiness and obesity, with no significant contribution of sleep fragmentation, hypoxaemia and apnoea recurrence. PMID- 12762358 TI - The role of mean inspiratory effort on daytime sleepiness. AB - This study has investigated the role of average maximum inspiratory effort in excessive daytime sleepiness in patients with obstructive sleep apnoea syndrome (OSAS) and upper airway resistance syndrome (UARS). Fifteen patients diagnosed with UARS and 32 patients with OSAS, with >5.5 h total sleep time (TST) during 8 h of nocturnal polygraphic recordings, were included in the study. Demographical data, polysomnographical data and data about daytime sleepiness, including Epworth sleepiness scale (ESS) and multiple sleep latency test (MSLT), were evaluated. In order to compute the average maximum inspiratory effort from oesophageal pressure (Poes) measurements, maximum Poes was obtained from 20 representative obstructive respiratory events (obstructive apnoeas, hypopnoeas or flow limitations) for each sleep stage in both supine and side positions. From Poes measurements during sleep, the increase in Poes (deltaPoes) during respiratory events was also calculated. The average maximum Poes, deltaPoes, respiratory disturbance index (RDI) and arousal index were significantly correlated with ESS in OSAS patients. In patients with UARS, the only significant correlation was obtained between average maximum Poes and ESS. The MSLT score did not show any significant correlation with arousal index, number of stage variations, RDI, average Poes, deltaPoes, minimum oxygen saturation (Sa,O2) and percentage of TST with an Sa,O2 <90% in both UARS and OSAS patients. The results of multiple regression analysis showed that average maximum Poes correlates best with the variance in ESS for OSAS patients. In conclusion, the data from this study indicate the possible important role of average inspiratory effort in determining subjective sleepiness in both obstructive sleep apnoea syndrome and upper airway resistance syndrome patients. PMID- 12762359 TI - A prediction rule to identify allocation of inpatient care in community-acquired pneumonia. AB - The current authors developed a new prediction rule based on the five risk classes defined by the Pneumonia Severity Index to identify allocation of inpatient care in community-acquired pneumonia. The decision to hospitalise in low-risk classes (I-III) was unquestionable, if the presence of one or more of the following were evident: arterial oxygen tension <8.0 kPa (60 mmHg), shock, decompensated coexisting illnesses, pleural effusion, inability to maintain oral intake, social problem, and lack of response to previous adequate empirical antibiotic therapy. The results at 18 months after implementation of this new prediction rule are reported in a series of 616 patients. The mortality rate was 0.5% in 221 patients treated as outpatients versus 8.9% in 395 patients treated as inpatients. Specific additional criteria for hospitalisation included in the prediction rule were present in 106 of the 178 low-risk patients treated as inpatients, whereas in the remaining 72, the decision to hospitalise was apparently unjustified by the prediction rule. These 72 patients showed a better outcome (significantly shorter hospitalisation, days on intravenous antibiotics, mortality, and complicated course) than high-risk patients and low-risk patients who met the additional specific criteria for deciding hospital admission. Therefore, admission in these low-risk patients might have been avoided by strict adherence to the new prediction rule. Another relevant finding was that the Pneumonia Severity Index alone did not identify all patients who needed to be admitted to the hospital. PMID- 12762360 TI - C-reactive protein levels in community-acquired pneumonia. AB - The diagnostic value of C-reactive protein (CRP) admission serum levels as an indicator of the aetiology of community-acquired pneumonia (CAP) was evaluated. A cohort of 1,222 patients with CAP was assessed. CRP levels were analysed in 258 patients with a single aetiological diagnosis. The mean CRP values in patients with pyogenic, atypical, viral and Legionella pneumophila pneumonia were: 16 mg x dL(-1), 13 mg x dL(-1), 14 mg x dL(-1) and 25 mg x dL(-1), respectively. CRP levels were not significantly different among patients outcome research team (PORT) groups (19 mg x dL(-1) in groups I-II, 16 mg x dL(-1) in group III and 16 mg x dL(-1) in groups IV-V). A cut-off point of 25 mg x dL(-1) had a sensibility, specificity, positive predictive value and negative predictive value of 0.6, 0.83, 0.3, and 0.94, respectively. After controlling for age and PORT score, the odds of having a CRP level >25 mg x dL(-1) was 6.9 times higher in patients with L. pneumophila pneumonia than in those with non-L. pneumophila pneumonia. Patients with Legionella pneumophila pneumonia had higher C-reactive protein levels than those with pneumonia of any other aetiology, independently of severity of infection. Being a cheap and readily available test, C-reactive protein may be a useful adjunctive procedure in the diagnosis of community acquired pneumonia. PMID- 12762361 TI - Definitions and types of work-related asthma: a nosological approach. AB - The workplace can trigger or induce asthma and cause the onset of different types of work-related asthma. Analysis of previous definitions of occupational asthma (OA) led to the conclusion that evidence of a direct causal relationship between workplace exposure and the development of asthma remains the key element for defining OA. Based on clinical features and pathophysiological mechanisms, the following conditions should be distinguished in the spectrum of work-related asthma: 1) immunological OA characterised by a latency period necessary to acquire immunologically induced sensitisation; 2) nonimmunological OA characterised by the rapid onset of asthma following single or multiple exposures to high concentrations of irritant compounds; 3) work-related asthma defined by exacerbation of symptoms in workers with pre-existing or coincident asthma; and 4) variant syndromes including eosinophilic bronchitis, potroom asthma, and asthma-like disorders caused by organic dusts. The issues and controversies relating to this approach are critically reviewed in order to stimulate the consensus development of operational definitions of work-related asthma. PMID- 12762362 TI - Coxiella burnetii pneumonia. AB - This report reviews the pulmonary and extrapulmonary manifestation of infections due to Coxiella burnetii. Q fever, a zoonosis, is due to infection with C. burnetii. This spore-forming microorganism is a small gram-negative coccobacillus that is an obligate intracellular parasite. The most common animal reservoirs are goats, cattle, sheep, cats, and occasionally dogs. The organism reaches high concentrations in the placenta of infected animals. Aerosolisation occurs at the time of parturition and infection follows inhalation of this aerosol. There are three distinct clinical syndromes of the acute form of the illness: nonspecific febrile illness, pneumonia, and hepatitis. The chronic form of Q fever is almost always endocarditis, but occasionally it is manifest as hepatitis, osteomyelitis or endovascular infection. The pneumonic form of the illness can range from very mild-to-severe pneumonia requiring assisted ventilation. Multiple round opacities are a common finding on chest radiography. Treatment with doxycycline or a fluoroquinolone is preferred. Susceptibility to macrolides is variable. In conclusion, Coxiella burnetii pneumonia should be considered when there is a suitable exposure history and when outbreaks of a pneumonic illness are being investigated. PMID- 12762363 TI - Pathophysiology and pharmacological treatment of pulmonary hypertension in acute respiratory distress syndrome. AB - Pulmonary hypertension (PH) is a characteristic feature of the acute respiratory distress syndrome (ARDS). The magnitude of PH has been shown to correlate with the severity of lung injury in patients with ARDS independently of the severity of associated hypoxaemia and has an adverse prognostic significance. Early in the histopathological evolution of ARDS, pulmonary vasoconstriction, thromboembolism and interstitial oedema contribute to the development of PH, although pulmonary vascular remodelling probably occurs eventually. Intravenous vasodilator agents lead to an increase in intrapulmonary shunting and systemic hypotension, which can limit their therapeutic use, and have not been shown to improve survival. By contrast, rapidly metabolised vasodilators administered by inhalation induce selective pulmonary vasodilatation and decrease shunting, but again do not appear to confer a survival benefit. Research aimed at further understanding the mechanisms that underlie pulmonary hypertension, a characteristic feature of the acute respiratory distress syndrome, are expected to provide improvements in pharmacological interventions for the treatment of pulmonary hypertension in the acute respiratory distress syndrome. PMID- 12762364 TI - Progressive dyspnoea in a 40-yr-old female. PMID- 12762365 TI - The best treatment for the first episode of primary spontaneous pneumothorax: an unanswered question. PMID- 12762366 TI - Carbon dioxide production during acetazolamide and medroxyprogesterone treatment. PMID- 12762367 TI - A tour in the desert. Veterinarians fill forward positions in Iraq campaign. PMID- 12762368 TI - The face of the veterinary profession. Educators wrestle with promoting racial diversity in the veterinary colleges. PMID- 12762369 TI - AVMA sets its legislative agenda for 108th Congress. PMID- 12762370 TI - What can you do to help? Tips for contacting your representatives in Congress. PMID- 12762371 TI - Veterinarians the 'first line of defense' in biologic attack. PMID- 12762372 TI - National pet health surveillance system in the works. PMID- 12762373 TI - Breed discrimination bites homeowners. Insurance companies dropping home insurance coverage for owners of large dog breeds. PMID- 12762374 TI - Disputes prevalence of Leishmania carriers in the United States. PMID- 12762375 TI - In defense of bats. PMID- 12762376 TI - Criticized veterinarian speaks out. PMID- 12762377 TI - Educational preparedness of veterinarians for foreign animal diseases. AB - Veterinary medical education in FADs has been and will continue to be critically important if veterinarians are expected to fulfill the profession's primary obligations to society--those of protecting our animals' health, conserving our animal resources, and promoting public health. It is imperative that curricula and instruction in veterinary schools and colleges provide the depth and breadth of knowledge and understanding necessary to prepare all veterinarians, including those in private practice, for their key role in defending against FADs. Development and implementation of governmental and military programs to diagnose, prevent, control, and eradicate FADs will require a dedicated cadre of public sector veterinarians who have a solid educational foundation in FADs and understand the contemporary issues and global challenges we face. Animal-related industries, associations, and organizations will increasingly rely on well educated veterinarians to help guide them in ways that will protect animals, clientele, consumers, and trading partners from effects of FADs. Agencies and organizations concerned with conservation of animal resources will require veterinary expertise necessary to prevent FADs in a multitude of animal species, including marine animals, wildlife, endangered species, zoologic specimens, and important genetic lines as well as our domestic companion and livestock species. Species affected by FADs also include human beings for those disease agents with zoonotic potential; thus, veterinary education also plays a key role in public health. PMID- 12762378 TI - What is your diagnosis? Sclerosis of the caudal vertebral body end plate of L7 and the cranial end plate of S1 and narrowing of the L7-S1 intervertebral disk space. PMID- 12762379 TI - Theriogenology question of the month. In utero infection of the doe by CVV. PMID- 12762380 TI - Evaluation of urine sulfated and nonsulfated bile acids as a diagnostic test for liver disease in dogs. AB - OBJECTIVE: To evaluate 3 methods for measuring urine bile acids (UBA) and compare their diagnostic performance with that of the serum bile acids (SBA) test and other routine screening tests in dogs with hepatic disorders. DESIGN: Prospective study. ANIMALS: 15 healthy dogs, 102 dogs with hepatic disorders, and 9 dogs with clinical signs of hepatic disorders that were found to have nonhepatic disorders. PROCEDURES: Blood and urine samples were collected from sick dogs and healthy dogs for serum biochemical analyses, and determination of concentrations of SBA and UBA. Urine samples were obtained from 15 healthy dogs to establish an upper cutoff value for UBA concentrations. The UBA were measured by use of a quantitative-linked enzymatic colorimetric method. Three analytical modifications were evaluated; 1 quantified only urine sulfated bile acids (USBA), 1 only urine nonsulfated bile acids (UNSBA), and 1 quantified both (USBA plus UNSBA). The UBA values were standardized with the urine creatinine concentration. RESULTS: The UNSBA-to-creatinine ratio and USBA plus UNSBA-to-creatinine ratio tests had the best diagnostic performance of the UBA tests; each had a substantially higher specificity, slightly higher positive predictive value, slightly lower negative predictive value, and lower sensitivity than the SBA test. These UBA-to creatinine values were positively correlated with SBA values. The USBA-to creatinine ratio had poor sensitivity, indicating a low rate of bile acid sulfation in dogs. CONCLUSIONS AND CLINICAL RELEVANCE: The UBA can be measured in dogs with sufficient repeatability and accuracy for clinical application. The UNSBA-to-creatinine ratio and USBA plus UNSBA-to-creatinine ratio identified dogs with hepatic disorders nearly as well as the SBA test. PMID- 12762381 TI - Use of a commercially available culture system for diagnosis of Tritrichomonas foetus infection in cats. AB - OBJECTIVE: To evaluate the efficacy of and optimize a commercially available culture system for sensitive and specific in-clinic culture of Tritrichomonas foetus from cat feces. DESIGN: Prospective study. SAMPLE POPULATION: Samples of freshly voided feces from 117 purebred cats and pure cultures of T. foetus obtained from a cat with chronic diarrhea. PROCEDURE: Optimal conditions for use of the culture system, such as quantity of fecal inoculum (0.025 to 0.2 g) and cultivation temperature (25 or 37 degrees C [98.6 or 77.0 degrees F]), were determined. Specificity of the system was examined by attempted culture of Giardia lamblia and Pentatrichomonas hominis. Sensitivity of the system to detect T. foetus was determined by inoculation of culture system pouches with serially diluted T. foetus suspensions with and without feces. RESULTS: Detection limit of the culture system was 1 and 1,000 T. foetus organisms without and with feces from cats, respectively. Optimal fecal inoculum was < 0.1 g of feces. At 37 degrees C, cultures yielded positive results in 24 hours; organisms remained viable for 1 to 6 days, and bacterial overgrowth was common. At 25 degrees C, cultures yielded positive results in 1 to 11 days; organisms were long-lived, and bacterial overgrowth was uncommon. Neither G. lamblia or P. hominis survived in the culture system. CONCLUSIONS AND CLINICAL RELEVANCE: The culture system was sensitive and specific for culture of T. foetus in feces of cats. Performance was optimal when test kits were inoculated with < or = 0.1 g of freshly voided feces and cultured at 25 degrees C. PMID- 12762383 TI - Semitendinosus muscle transfer flap for external anal sphincter incompetence in a dog. AB - A 5-year-old sexually intact female Yorkshire Terrier was referred with a history of fecal incontinence of at least 2 years and chronic intermittent colitis. The external anal sphincter to the left of the anus was intact; the external anal sphincter was not detectable to the right of the anus. To repair the defect, the semitendinosus muscle was isolated and severed 2 cm proximal to its insertion on the tibia. Care was taken to preserve the integrity of the vasculature and nerve supply in the proximal third of the muscle body. The body of the muscle was passed around the ventral and right aspects of the rectum; the cut end was secured with simple interrupted sutures dorsal to the levator ani and coccygeus muscles to simulate the external anal sphincter. After surgery, the dog could defecate normally. Absence of a portion of the external anal sphincter may be congenital or the result of anorectal trauma, rectal prolapse, severe perineal disease, or surgical resection. The use of a semitendinosus muscle flap for treatment of fecal incontinence secondary to sphincter incompetence in dogs may be a viable alternative to euthanasia. PMID- 12762382 TI - Fungal rhinitis and sinusitis in three cats. AB - Localized infection of the nasal or paranasal cavities caused by Aspergillus spp or Penicillium spp was diagnosed in 3 cats. Clinical signs included chronic mucopurulent nasal discharge, epistaxis, and mandibular lymphadenopathy. Rhinoscopic and diagnostic imaging findings were compatible with severe inflammation of the nasal mucosa and destruction of the turbinates. Fungal plaques were observed rhinoscopically in 2 cats, and histologic examination of biopsy specimens revealed fungal colonies with surrounding inflammatory infiltrates in all 3. Results of fungal culture were negative for all 3 cats. Results of serum immunoelectrophoresis for antibodies against Aspergillus spp were positive in 2 cats. Treatment with itraconazole was effective in controlling clinical signs in 1 cat, but hepatotoxicosis developed. A single intranasal infusion of clotrimazole subsequently led to long-term resolution of clinical signs in this cat. Localized aspergillosis-penicilliosis is clinically indistinguishable from other pathologic conditions of the nasal and paranasal cavities in cats and should be considered when examining cats with chronic nasal discharge. PMID- 12762384 TI - Risk factors for sterile hemorrhagic cystitis in dogs with lymphoma receiving cyclophosphamide with or without concurrent administration of furosemide: 216 cases (1990-1996). AB - OBJECTIVES: To determine incidence and identify predisposing factors for sterile hemorrhagic cystitis (SHC) in dogs with lymphoma that were treated with cyclophosphamide and to evaluate whether furosemide administered i.v. concurrently with cyclophosphamide decreased the incidence of SHC. DESIGN: Retrospective study. ANIMALS: 216 dogs with lymphoma. PROCEDURE: Medical records of dogs with lymphoma that received cyclophosphamide chemotherapy in accordance with 1 of 2 protocols, with or without concurrent i.v. administration of furosemide, were examined. Data for the 2 groups were analyzed to determine the incidence and predisposing factors (age, breed, sex, weight, previous or preexisting disease, previous or preexisting urinary tract infection, neutropenia, azotemia, dose, and number of cyclophosphamide treatments) for cyclophosphamide-associated SHC. RESULTS: Cyclophosphamide-associated SHC developed in 12 of 133 (9%) dogs that had not received concurrent administration of furosemide and cyclophosphamide treatments; of the 83 dogs that had received furosemide, only 1 (1.2%) developed SHC. Dogs receiving cyclophosphamide and furosemide concurrently were significantly less likely to develop SHC than dogs that did not receive furosemide. Dogs with previous or preexisting immune mediated disease were significantly more likely to develop cyclophosphamide associated SHC. CONCLUSIONS AND CLINICAL RELEVANCE: Analysis of results suggested an association between i.v. administration of furosemide concurrently with cyclophosphamide and decreased incidence of cyclophosphamide-associated SHC. Incidence of cyclophosphamide-associated SHC was similar in treated dogs that did not receive concurrent furosemide to that observed for other studies in which cyclophosphamide was administered orally. Cyclophosphamide-associated SHC appeared to develop early during the course of chemotherapy when furosemide was not administered concurrently with cyclophosphamide. PMID- 12762385 TI - Outcome of epidural catheterization for delivery of analgesics in horses: 43 cases (1998-2001). AB - OBJECTIVE: To determine reasons for epidural catheter placement among horses examined at a veterinary teaching hospital, efficacy of epidural administration of analgesics, duration of catheter placement, reasons for catheter removal, and complications encountered. DESIGN: Retrospective study. ANIMALS: 43 horses. PROCEDURE: Medical records were reviewed. RESULTS: A total of 50 epidural catheters were placed in the 43 horses. Underlying conditions included fractures, lacerations, septic arthritis, myositis, perineal injuries, and cellulitis. Horses ranged from 2 to 21 years old and weighed between 365 and 795 kg (803 and 1,749 lb). Median duration of catheter placement was 96 hours (range, 1.5 to 480 hours). The response to epidural drug administration was reported as positive in 34 horses and negative in 4. There was no apparent response in 2 horses, and response could not be determined in 3. Three temporary patient-related complications associated with epidural catheter administration were observed. Technical problems associated with the epidural catheters included dislodgement of the catheter itself (7 catheters) or of the adapter or filter (5), obstruction (5), and leakage (5). Twenty-two catheters were removed because of resolution of the underlying condition, and 10 were removed because of complications. For 6 catheters, the reason for catheter removal was not recorded. The remaining 12 catheters were in place when the horses were euthanatized. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that epidural catheterization can be used successfully for repeated epidural delivery of analgesics and anesthetics in horses with various clinical conditions. Complications associated with epidural catheters or epidural drug administration were infrequent and transient. PMID- 12762386 TI - Evaluation of shedding of bovine herpesvirus 1, bovine viral diarrhea virus 1, and bovine viral diarrhea virus 2 after vaccination of calves with a multivalent modified-live virus vaccine. AB - OBJECTIVE: To determine whether bovine herpesvirus 1 (BHV1), bovine viral diarrhea (BVDV) virus 1 (BVDV1), or BVDV 2 (BVDV2) were shed after parenteral administration of a multivalent modified-live virus vaccine. DESIGN: Prospective study. ANIMALS: 28 healthy beef calves and 4 healthy pregnant beef cows. PROCEDURE: A commercially available modified-live virus multivalent vaccine was administered to steers and heifers (n = 18) that were seronegative to BHV1, BVDV1, and BVDV2. Four seronegative pregnant control cows were held in contact with the vaccinated calves for 103 days. Unvaccinated calves (n = 10) were held as controls in a separate double-fenced pen. Seroconversion was monitored by determining serum neutralization titers after vaccination. Viral shedding and viremia were assessed via analysis of nasal swab specimens and blood by use of polymerase chain reaction (PCR) and reverse transcriptase-PCR assays and virus isolation. RESULTS: A transient BVDV1 viremia was detected in most vaccinated calves 3 to 10 days after vaccination. All vaccinated calves seroconverted to BVDV1 and BVDV2. Seventeen of 18 vaccinated calves seroconverted to BHV1. Viral shedding was not detected in the vaccinated calves. All control cattle remained seronegative to BHV1, BVDV1, and BVDV2 throughout the study. CONCLUSIONS AND CLINICAL RELEVANCE: Shedding of BHV1, BVDV1, and BVDV2 after vaccination was either nonexistent or undetected and did not result in transmission of BHV1, BVDV1, or BVDV2 vaccine viruses to pregnant contact control cows. PMID- 12762387 TI - Use of a polymerase chain reaction assay to detect bovine herpesvirus type 2 DNA in skin lesions from cattle suspected to have pseudo-lumpy skin disease. AB - Beef cattle from a herd in north Alabama were examined because of an outbreak of nonfatal skin disease characterized by discrete circumscribed areas of inflammation that developed on the skin from the neck to the hips. Areas of inflammation, which tended to be superficial, underwent necrosis and scabbed over. The scabs eventually dropped off leaving discrete, round, whitish, hairless lesions that were 1.2 to 2.5 cm diameter. Because clinical signs were consistent with those expected with pseudo-lumpy skin disease (PLSD) caused by bovine herpesvirus type 2 (BHV-2), samples from 16 representative animals were submitted for BHV-2 testing. All 16 animals were seropositive for BHV-2, but the virus could not be isolated from skin biopsy specimens or buffy coat samples. Results of a polymerase chain reaction assay incorporating primers designed to amplify 2 DNA sequences from BHV-2 were positive for 3 of the 10 cattle, suggesting that skin lesions in these cattle were a result of PLSD. Our findings suggest that PLSD may be more common and widespread in the United States than suggested by the frequency with which BHV-2 has been isolated from cattle with PLSD-like skin lesions. PMID- 12762388 TI - Spiral colon bypass in a geriatric Vietnamese potbellied pig. AB - An 8-year-old potbellied pig was evaluated for anorexia, decreased fecal production, signs of depression, inappetence, and abdominal distension. During hospitalization, a tooth root impaction and abscess were diagnosed, and abdominal radiography revealed severely distended, gas-filled large and small intestines. Exploratory laparotomy revealed a stricture of the proximal centripetal loop of the spiral colon and megacolon of the proximal portion of the spiral colon and cecum. A side-to-side spiral colon anastomosis was performed to create a partial bypass of the spiral colon. The success of this procedure suggests that spiral colon bypass is a treatment option for spiral colon stricture formation in potbellied pigs. Spiral colon stricture formation should be considered as a differential diagnosis in geriatric potbellied pigs that are anorectic, have abdominal distension, and have decreased fecal production. PMID- 12762389 TI - Avian macrophage and immune response: an overview. AB - Macrophages belong to the mononuclear phagocytic system lineage. This cell type is unique in that it is a crucial player in both the innate and adaptive immune responses. The material described in this overview is a brief description of what I presented as a World's Poultry Science Association-sponsored lecture at the annual meetings of the Poultry Science Association in 2002. Therefore, I have not attempted to present an up-to-date review of literature on this topic. Rather, I have summarized some salient research accomplishments made by our research group over the years in the area of avian macrophage biology and function. PMID- 12762390 TI - Genetic markers associated with antibody response kinetics in adult chickens. AB - A linkage disequilibrium approach with microsatellites was employed to investigate QTL affecting immune response. Highly inbred males of two MHC congenic Fayoumi chicken lines were mated with highly inbred G-B1 Leghorn hens. Adult F2 hens (n = 158) were injected twice with SRBC and fixed Brucella abortus (BA). Agglutinating antibody titers were measured. Secondary phase parameters of maximum titers (Ymax) and time (Tmax) needed to achieve Ymax were estimated from postsecondary titers by using a nonlinear regression model. A three-step genotype strategy (DNA pooling, selective genotyping, and whole population genotyping) was used to identify microsatellite markers that are associated with immune response to SRBC and BA. The linkage distances between adjacent markers in the F2 population were estimated by Crimap. The QTL affecting immune response to SRBC and BA were detected based on F statistic by interval mapping. A total of five significant QTL, as determined by a permutation test, were detected at the 5% chromosome-wise level on Chromosomes 3, 5, 6, and Z. Two (Chromosome 3 and 6) of five QTL were significant at the 1% chromosome-wise level. The variance explained by the QTL ranged from 6.46 to 7.50%. The results suggest that regions on Chromosomes 3, 5, 6, and Z contain QTL that affect antibody kinetics in the hen. PMID- 12762391 TI - Correlated responses to long-term selection for clutch length in dwarf brown-egg layers carrying or not carrying the naked neck gene. AB - Two dwarf brown-egg layer lines, differing in their genotype for the naked neck gene (NA), line L2 (NA*NA/*NA) and line L1 (NA*N/*N), have been selected for 16 generations for increased average clutch length. A control line from the same base population, dwarf and segregating for the NA gene, was maintained by random mating. Genetic parameters were estimated by a multivariate derivative-free restricted maximum likelihood procedure, and the NA gene effect was estimated within the unselected control line. The studied traits included clutch traits, egg production traits, abnormal eggs, egg weight at 36 wk, and BW at 42 wk. The average clutch length, egg number, and maximum clutch length were normalized using the Box-Cox transformation. In response to 16 generations of direct selection for increased average clutch length, other egg production traits, such as laying rate and total egg number, have been indirectly improved in a dwarf layer genetic background. The estimated heritabilities were 0.406 to 0.424 for transformed average clutch length (TCL), 0.373 to 0.411 for transformed egg number (TEN), 0.529 to 0.559 for age at first egg (AFE), 0.275 to 0.282 for laying rate (LR), 0.455 for dutch number (CN), and 0.319 for the number of double yolked eggs (DYEN). The TCL had high genetic correlations with TEN (0.777), LR (0.863), maximum clutch length (0.902), and CN (-0.845). Selection for increased average clutch length was an effective method for increasing egg production. Line L2 showed a higher egg weight than L1, which indicates that the combined effect of NA and DW genes was favorable to maintain egg weight when egg number could be improved. Line L1 showed a higher number of DYEN, suggesting that the regulation of follicular maturation was changed in this line. PMID- 12762393 TI - Poultry production: a model for developing interactive Internet-based distance education. AB - Over the last several decades, many poultry science programs have merged with other departments, but the poultry industry has undergone tremendous expansion worldwide, leading to a growing instructional void with regard to poultry production information. The objective of this project was to address the demand for information by developing two Web-based poultry production courses that cover management of broilers, turkeys, breeders, and layers. The Internet was chosen as the platform because it is asynchronous and may be accessed from any connected site around the world. To be effective, web-based courseware must be theoretically grounded and interactive, but university-level web-based distance education courses often fail to meet these standards. During courseware development, the impact of instructional techniques and technologies on interactivity and learning outcomes was explored. A content expert, an instructional designer, and a graphic artist carefully reviewed a variety of instructional techniques to increase interactivity. Concept mapping was chosen because it has been shown to be a superior learning tool for enhancing the exchange of ideas and knowledge between instructors, students, and content. A unique instructional interface was established that includes threaded e-mail discussion, thought questions, animation, hypertext, rollover interactions, video clips, and concept mapping exercises. Results indicate that the integration of concept mapping into web-based learning environments successfully increased interactivity and learning outcomes. PMID- 12762392 TI - Effect of two candidate genes on the Salmonella carrier state in fowl. AB - Selection for increased resistance to Salmonella carrier-state (defined as the persistency of the bacteria 4 wk after inoculation) could reduce the risk for the consumer of food toxi-infections. The effects of two genomic regions on chromosomes 7 and 17 harboring two genes, NRAMP1 (SLC11A1) and TLR4, known to be involved in the level of chicken infection 3 d after inoculation by Salmonella were thus tested on a total of 331 hens orally inoculated at the peak of lay with 10(9) bacteria. The animals and their parents were genotyped for a total of 10 microsatellite markers mapped on chromosomes 7 and 17. Using maximum likelihood analysis and interval mapping, it was found that the SLC11A1 region was significantly involved in the control of the probability of spleen contamination 4 wk after inoculation. Single nucleotide polymorphisms (SNP) within the SLC11A1 and TLR4 gene were tested on those animals as well as on a second batch of 279 hens whose resistance was assessed in the same conditions. As the former was significantly associated with the risk of spleen contamination and the number of contaminated organs, SLC11A1 appears to be involved in the control of resistance to Salmonella carrier state. The involvement of the TLR4 gene was also highly suspected as a significant association between SNP within the gene, and the number of contaminated organs was detected. PMID- 12762394 TI - Effects of egg storage time on spread of hatch, chick quality, and chick juvenile growth. AB - A total of 1,800 incubating eggs produced by a commercial flock of Cobb broiler breeders was used to determine the effects of storage duration (3 or 18 d) on spread of hatch and chick quality. Chick relative growth (RG) at the end of 7 d of rearing was also determined as a measure of the chick performance. Chick quality was defined to encompass several qualitative characteristics and scored according to their importance. Eggs stored for 3 d hatched earlier than those stored for 18 d (P < 0.05). Hatching was normally distributed in both categories of eggs, and the spread of hatch was not affected by storage time (P = 0.69). Storage duration of 18 d reduced the percentage of day-old chick with high quality as well as average chick quality score (P < 0.05). RG varied with length of egg storage, quality of day-old chick, and the incubation duration (P < 0.05). Eighteen-day storage of eggs not only resulted in longer incubation duration and lower quality score but also depressed RG. Chick quality as defined in this study was correlated to RG and storage time. It was concluded that day-old chick quality may be a relatively good indicator of broiler performance. The results suggest however that in order to improve performance prediction power of chick quality, it would be better to define it as a combination of several qualitative aspects of the day-old chick and the juvenile growth to 7 d. PMID- 12762395 TI - Early T-maze behavior and broiler growth. AB - In the laboratory and at a commercial farm in Argentina, chicks that showed a short latency to exit a T-maze (LEX) (HP, high performance) gained more weight than those with a longer LEX (LP, low performance). The present study re-examined this relationship and evaluated additional T-maze measures using broilers reared in quasi-intensive, environmentally controlled conditions. In the T-maze, a mirror strategically placed at the end of a corridor stimulates the test chick to leave the isolation chamber (start box) and to move toward its reflection. Having done so, it can then see other birds and thereby be further stimulated to completely exit the maze. However, when a test bird hesitates or stops at the mirror, its performance categorization may be confounded. Herein, the T-maze performance of 3-d-old broiler chicks was assessed using three different measurements: 1) the latency to exit the start box (LEB), 2) the latency to reach the mirror section of the maze (LRM), and 3) LEX. The fastest (top 25%) and slowest (bottom 25%) of the birds within a sex and within each of the three T maze criteria were classified as HP and LP chicks, respectively. The relationships between these measures and body weight were examined in 4-, 42-, and 56-d-old males and females. Irrespective of the T-maze classification criteria used, HP and LP chicks had similar body weights at 4 d of age. However, chicks classified as HP according to LEB or LRM measures were heavier at 42 and 56 d of age than LP ones (P < 0.02 for all comparisons). These differences were apparent in both sexes. On the other hand, there were no detectable differences in body weight at the latter ages between chicks categorized as HP or LP according to their LEX scores. The present results indicate that 1) broiler chicks that exit the T-maze start box and reach the mirror quickly subsequently grow faster than their slower LP counterparts and 2) LEB and LRM are better predictors of growth than the LEX value used in previous studies. PMID- 12762396 TI - Evaluation of nonfeed removal methods for molting programs. AB - An experiment was conducted using 336 White Leghorn hens (60 wk of age) randomly assigned to one of four treatments that consisted of feed removal for 4 or 10 d or no feed removal with ad libitum access to 95% corn or 95% wheat middlings molt diets that contained supplemental minerals and vitamins. At the end of the 4- or 10-d feed removal period, hens on these treatments were provided with the corn molt diet for 24 or 18 d, respectively. Hens on the corn or wheat middlings treatments were fed the diets for 28 d. At d 28, hens on all treatments were fed a corn-soybean meal (16% CP) layer diet for 40 wk (64 to 104 wk of age). Both feed removal and the wheat middlings treatments resulted in total cessation of egg production within 8 d. Egg production of hens fed the corn molt diet had decreased to 3% by d 28. Body weight loss for hens fed the corn or wheat middlings diet was approximately 15 and 8% at d 28, respectively. Hens fed the wheat middlings diet returned to production slightly faster than hens on the other treatments. Postmolt egg production and egg mass (wk 5 to 44) were generally higher for the wheat middlings and 10-d feed removal treatments than for the corn or 4-d feed removal treatments. There were no consistent differences in mortality, egg weight, egg specific gravity, feed efficiency, and layer feed consumption among treatments. This research indicates that diets with high corn or wheat middlings, particularly wheat middlings, are effective nonfeed removal methods for molting hens. PMID- 12762397 TI - Evaluation of a method of ultraviolet light sanitation of broiler hatching eggs. AB - Sanitation of hatching eggs is an important area of research due to the need for an effective, economical, and safe method of egg sanitation. Improved hatching egg sanitation is an important part of an overall pathogen reduction program within integrated poultry operations. This must be accomplished without disturbing the cuticle of the egg, which can decrease hatchability. The ability of ultraviolet (UV) light to kill bacteria on eggshell surfaces has been well documented. To accomplish the task of treating the eggs in a method that could be commercially implemented, a cabinet was constructed in which ultraviolet lamps were placed. A conveyor system was used to carry a plastic hatching egg flat containing 42 eggs through the cabinet for a period of 3 or 4 min. Ultraviolet intensities within the cabinet reached a maximum of 14 mW/cm2. Experiments were conducted to test the impact of UV light (254 nm) exposure of hatching eggs on aerobic plate counts (APC), inoculated Salmonella typhimurium and inoculated Escherchia coli. In the first three experiments, seven eggs were sampled from a flat passed through the UV chamber. Ultraviolet-treated eggs compared to untreated eggs had APC reductions of 1.3 log, S. typhimurium had a 4 log reduction, and E. coli had a 4 to 5 log reduction. Laboratory trials were also conducted to test the effects of UV irradiation on the cuticle of the egg and hatchability. No significant differences for eggshell conductance or hatchability were found between UV-treated and control eggs. From these trials, it can be concluded that UV irradiation of hatching eggs in a prototype irradiation cabinet can effectively reduce aerobic and pathogenic bacteria on eggshell surfaces without affecting eggshell conductance or hatchability. PMID- 12762398 TI - Effects of lentinan on broiler splenocyte proliferation, interleukin-2 production, and signal transduction. AB - Lentinan (LEN) is an intensively studied beta-glucan that has been shown to have immunostimulating and antitumor functions. However, it currently is not used in veterinary practice and animal production. This study was conducted to investigate the impacts of LEN on broiler splenocyte proliferation, interleukin-2 (IL-2) production, and to explore its effects on receptor signal transduction by determining signaling molecules, including nitric oxide (NO), cytosolic-free Ca2+, cytosolic cyclic adenosine monophosphate (cAMP), and cyclic guanosine monophosphate (cGMP). The results showed that compared to the control all three measured doses, including 40, 80, and 160 microg/mL LEN, increased splenocyte proliferation and IL-2 production (P < 0.05). Lentinan increased splenocyte NO production and cytosolic-free calcium concentration in a dose-dependent manner (P < 0.001). After incubating splenocytes for 20 min, LEN increased intracellular cAMP and cGMP concentrations at a dose of 80 and 160 microg/mL, respectively. However, after incubating the splenocytes for 60 min, LEN had no effect on cytosolic cAMP and cGMP concentration irrespective of the LEN dosage. Those results indicated that LEN had immunostimulatory effects on splenocytes by increasing splenocyte proliferation and IL-2 production and by activating splenocyte receptor signal transduction. PMID- 12762399 TI - Esophageal tonsil: a novel gut-associated lymphoid organ. AB - The esophageal tonsil of the chicken is a novel, significant element of the gut associated lymphoid tissue (GALT). Its stable location and histological organization fulfills the meaning of the term "tonsil." The six-to-eight-isolated tonsillar units are located at the border of the esophagus and the proventriculus. The number of tonsillar units is identical with that of the esophageal folds. Each tonsillar unit consists of a crypt lined by lymphoepithelium and surrounded by dense lymphoid tissue, which is organized into T- and B-dependent regions, like peripheral lymphoid organs. The excretory ducts of the mucosal glands of the esophagus are frequently involved in the formation of the lymphoepithelium. The esophageal tonsil is anatomically located cranial to the stomach, unlike the other parts of the GALT. Therefore, it is continuously exposed to undigested environmental antigens, allergens, food, and infectious agents. To develop effective oral vaccines, the existence of the esophageal tonsil has to be taken into account. PMID- 12762400 TI - Pulmonary and hematological inflammatory responses to intravenous cellulose micro particles in broilers. AB - When injected intravenously, cellulose micro-particles become lodged in pulmonary arterioles. The current study investigated the systemic and pulmonary inflammatory responses triggered by cellulose micro-particles at 3, 24, and 48 h postinjection in 6-wk-old broilers. Proportions and concentrations of circulating white blood cells were assessed in saline-injected (control group) and cellulose injected (particle group) birds. Hematoxylin-eosin (HE)-stained cross-sections of the lungs were used to count the number of granuloma/lymphocyte aggregates, which is indicative of the severity of the inflammatory response to the trapped particles. The cellular components of the aggregates were identified by immunohistochemical staining of frozen cross sections of the lungs. Results showed that cellulose micro-particles trapped in the pulmonary vasculature initiated a dynamic, localized inflammatory response within the surrounding lung parenchyma. Monocytes and basophilic granulocytes closely surrounded the particles. CD4, CD8, TCR1, TCR2, and TCR3 subsets of T cells and B cells were present in the outer rim of the granuloma/lymphocyte aggregates. Circulating total white blood cell (WBC, leukocytes) concentrations were similar in both groups at all times postinjection, whereas at 48 h post-injection the percentages of eosinophils and basophils among circulating WBC were higher in the particle group than in the control group (P < or = 0.05). The circulating monocyte concentration also increased within 24 h postinjection (P < or = 0.05). These observations demonstrate that cellulose micro-particles trapped in the pulmonary vasculature initiated acute focal inflammatory responses in the lungs and that the proportions of WBCs in the blood are modulated within 48 h postinjection. PMID- 12762401 TI - Effects of delayed feed intake on body, intestine, and muscle development in neonate broilers. AB - The short-term effects of delayed feeding (DF) for 2 d posthatching were measured in neonate chicks and compared to early feeding (EF). Chicks from 10 independent families were used in this study to determine whether genetic background control of growth may be influenced by EF and DF. Early feeding maintained significant interfamily body weight variations from hatch to 4 d of age, whereas there were no significant differences from 1 d of age when feeding was delayed to 48 h posthatching. These results suggest that posthatching feeding delay may distort genetic selection by masking the expression of genetic potential and disturbing the estimation of chick breeder value. In DF chicks, overall body growth was delayed until the beginning of feeding and body weight at 6 d of age was 25% lower than EF chicks. Availability of feed after the fasting period was not sufficient to compensate for the retardation of weight gain in either body weight or in intestine and breast muscle weight. However, initiation of intestine growth in DF chicks occurred from 1 d of age despite the lack of feeding, whereas feed intake was essential to enhance muscle growth. The potential for protein synthesis was lower in DF than in EF chicks during the first 2 d posthatching (P < 0.001) and then reached similar values after feed intake. These results confirm that initiation of growth in neonate chicks is improved by earlier feeding after hatching. Awareness of changes in overall body weight caused by posthatching food deprivation, especially in the intestine and muscle might help in the development of new diets which could minimize retardation of body weight gain in chicks. PMID- 12762402 TI - High available phosphorus corn and phytase in layer diets. AB - High available phosphorus corn (HAP) developed using the low phytic acid 1-1 (lpal-1) allele of the corn LPA1 gene containing 0.27% P, with 0.17% nonphytate P (NPP), was compared to near isogenic normal corn (LPA1), which contained 0.23% P and 0.05% NPP. Five levels of NPP from either HAPC or normal corn (0.40, 0.35, 0.30, 0.25 and 0.20% + 300 phytase units (FTU)/kg microbial phytase) were combined in a 2 x 5 factorial experiment for a total of 10 dietary treatments. Each dietary treatment was fed to eight replicate cages with five Hy-Line W-36 hens per replicate cage from 20 to 40 wk of age. Feed consumption and egg production were not significantly affected by dietary NPP level or corn type. Feed conversion ratio (g feed:g egg mass) was improved at the 0.35% NPP level (1.856) compared to the other levels of NPP--0.40, 0.30, 0.25, and 0.20% + phytase having feed conversion ratios of 1.872, 1.905, 1.930, and 1.898, respectively. Egg weight and egg mass decreased significantly as dietary NPP decreased; diets with 0.20% NPP plus phytase had equal egg mass to the 0.35 and 0.40% NPP diets. A significant corn type x NPP interaction effect was observed for egg weight, such that within the HAP corn diets, egg weight decreased more markedly at the 0.25% NPP levels compared to the normal corn 0.25% NPP diets. Specific gravity was not affected by dietary treatment, but percent dry shell was improved at the lower AP levels and with phytase treatment. Dietary NPP level and corn type had no significant effect on bone ash. Excreta levels of total phosphorus decreased significantly as dietary NPP decreased and were lower in the HAP corn excreta compared to normal corn excreta. Total P, Ca, Zn, Cu, and Mn retention were significantly affected by NPP level and corn type. HAP corn reduced Ca, Zn, Cu, and Mn retention compared to normal corn; this negative effect was alleviated by phytase supplementation to HAP corn diets. HAP corn allowed less dicalcium phosphate supplementation in layer diets compared to normal corn while supporting equal egg production. Phytase supplementation of low NPP diets had no significant positive effects on egg production parameters in either corn type diets. PMID- 12762403 TI - The effects of conjugated linoleic acids or an alpha-glucosidase inhibitor on tissue lipid concentrations and fatty acid composition of broiler chicks fed a low-protein diet. AB - The effects of dietary conjugated linoleic acid (CLA) and an alpha-glucosidase inhibitor--Bay g 5421--on tissue lipid concentrations and fatty acid composition were investigated in male broiler chicks fed a low-protein diet. The trial comprised six isoenergetic broiler diets. One diet (high-protein diet) was used as a control diet and contained 230 g crude protein per kilogram. The other five diets were low-protein diets (180 g crude protein/kg) without additional supplement, or supplemented with 20 or 40 g/kg of a CLA-enriched oil, or supplemented with 50 or 100 mg of Bay g 5421/kg. Broilers fed the low-protein diets had higher concentrations of triglycerides and cholesterol in the liver than broilers fed the high-protein diet. Broilers fed low-protein diets supplemented with CLA had a higher cholesterol concentration in plasma, lower hepatic triglyceride concentrations, higher relative concentrations of saturated fatty acids (SFA), and lower relative concentrations of monounsaturated fatty acids (MUFA) in lipids of liver, thigh muscle, and adipose than broilers fed low protein diets without CLA supplement. Broilers fed the low-protein diets supplemented with Bay g 5421 had lower concentrations of triglycerides and cholesterol in the liver and lower concentrations of triglycerides in plasma than broilers fed the low-protein diet without supplementation. Moreover, broilers fed diets with Bay g 5421 had higher relative concentrations of polyunsaturated fatty acids and lower relative concentrations of MUFA in lipids of liver, muscle, and adipose tissue. The results show that supplementation of the low-protein diets with CLA and Bay g 5421 affected tissue lipid concentrations and fatty acid composition in broilers, which are of practical relevance with regards to meat quality. PMID- 12762404 TI - Effects of microflora status, dietary bile salts and guar gum on lipid digestibility, intestinal bile salts, and histomorphology in broiler chickens. AB - The aim of the present study was to reexamine the effects of intestinal viscosity, microflora, and bile salts, and their interactions in order to clarify the mechanisms that explain the effect of intestinal viscosity on lipid digestibility, especially those that could involve microflora. Effects of intestinal viscosity, microflora status, and dietary bile salts on lipid digestibility, intestinal bile salts, and intestinal histomorphology were tested in a 2 x 2 x 2 factorial experiment. The effect of microflora status was examined by comparing conventional chickens to chickens with limited microflora obtained by rearing birds in sterilized conditions. Viscosity and dietary bile salts were tested with guar gum (0 or 0.5% in diets) and sodium taurocholate (0 or 0.3% in diets), respectively. Guar gum was autoclaved and added to the gamma-irradiated diets by mixing inside the sterile isolators. The intestinal concentration of lactic acid and cecal concentration of short-chain fatty acids were both very low in birds with limited microflora compared to conventional birds (P = 0.0001). Chickens with limited microflora had higher gain:feed ratios (P = 0.002), higher fecal lipid digestibility (P = 0.0001), more intestinal conjugated bile salts (P = 0.0001), less intestinal unconjugated bile salts (P = 0.0001), and their gastrointestinal compartments were smaller (P < 0.05) than those of conventional chickens. Addition of bile salts in diets increased the lipid digestibility (P = 0.0001) with a more pronounced effect in conventional birds (P = 0.0001) and in birds fed on guar gum diets (P = 0.002). Feeding the guar gum diets increased the intestinal supernatant viscosity (P = 0.0001) for both microflora status. Guar gum addition increased (P < 0.05) the lactic acid concentration in the small intestine of conventional chickens. Guar gum decreased the fecal lipid digestibility (P = 0.0001) and the intestinal conjugated bile salts (P = 0.0001) for both microflora status. However, the magnitude of lipid digestibility decrease due to guar gum was lower (P = 0.05) in birds with limited microflora than in conventional birds, and the negative effect of guar gum on intestinal conjugated bile salts was more pronounced (P = 0.02) in birds with limited microflora. Bile salt addition reduced the negative effect of guar gum on lipid digestibility (P = 0.02) for both microflora status. The mean lipid digestibilities were negatively correlated (P = 0.0001) with the ratio [Ln(viscosity)/total bile salt] measured in intestinal digesta. Except for gizzard, size of digestive compartments and villus heights increased (P < 0.05) with feed efficiency and digestibility decreased. In conclusion, the results provided evidences that the negative effect of guar gum on lipid digestibility was mainly mediated by its direct effect on intestinal bile salt concentration and efficiency. The small interaction between guar gum and microflora status effects on lipid digestibility had to be accounted for by the low basal level of intestinal bile salts in conventional chickens. PMID- 12762405 TI - Requirement of tryptophan in relation to the supply of large neutral amino acids in laying hens. AB - The present study was undertaken to find out whether the tryptophan requirement of laying hens is influenced by the supply of large neutral amino acids (LNAA). A factorial experiment was performed in which the dietary tryptophan concentration was varied at six different levels (1.0, 1.25, 1.5, 1.75, 2.0, and 2.5 g tryptophan/kg diet). As the second factor, the dietary concentrations of LNAA (isoleucine, valine, leucine, phenylalanine, and tyrosine) were varied at two levels. The first level provided an adequate supply of these amino acids; at the second level the concentrations of these amino acids were 40% higher than at the first level. The tryptophan requirement was estimated by a broken-line model and an exponential model of regression analysis. The tryptophan intake required for optimum (100% of maximum in the broken-line model, 95% of the maximum in the exponential model) egg production and daily egg mass was lower in hens fed the diets with high LNAA concentrations (145 and 155 mg/hen per day, respectively, in average of both models) than in hens fed the diets with adequate concentrations of LNAA (184 and 198 mg/hen per day, respectively, in average of both models). In contrast, the tryptophan requirement for optimum BW gain was lower in hens fed the diets with adequate LNAA concentrations (178 mg tryptophan per day) than in hens fed the diets with a high concentration of LNAA (212 mg tryptophan per day). In conclusion, the study suggests that an interaction between dietary LNAA and tryptophan exists in laying hens. PMID- 12762406 TI - Comparison of broiler performance when fed diets containing grain from YieldGard (MON810), YieldGard x Roundup Ready (GA21), nontransgenic control, or commercial corn. AB - This 42-day experiment was undertaken to compare the nutritional value of insect protected corn event MON810 (YieldGard) and YieldGard x herbicide-tolerant corn event GA21 (Roundup Ready) to their nontransgenic controls as well as four different commercial reference corns, when fed to growing Cobb x Cobb broilers. A randomized complete block design was used, and each treatment was replicated with five pens of males and five pens of females with 10 broilers per pen. Broilers were fed approximately 55% wt/wt corn during the first 20 d and approximately 60% wt/wt corn thereafter. The corn component of diets fed to broilers was supplied entirely with grain from the eight hybrids included in the experiment. Final live weights averaged 2.09 kg/bird fed YieldGard corn and 2.15 kg/bird fed YieldGard x Roundup Ready corn and were not different (P > 0.05) from final weights for birds fed control or commercial corn. Feed conversion was not affected (P > 0.05) by YieldGard (1.72) or YieldGard x Roundup Ready (1.77) corn feeding when compared with the feeding of other corn diets. Chill weights, fat pad, thigh weights, and wing weights were not affected by diets (P > 0.05). Differences (P < 0.05) were noted for breast and drum weights across treatments. Broilers overall performed consistently and had similar carcass yield and meat composition when fed diets containing YieldGard (event MON810) or YieldGard (event MON810) x Roundup Ready (event GA21) as compared with their nontransgenic controls and commercial diets. PMID- 12762407 TI - Long-term supplementation of various dietary lipids alters bone mineral content, mechanical properties and histological characteristics of Japanese quail. AB - The purpose of this study was to investigate the effects of long-term supplementation of fat in the diets on the fatty acid composition, chemical, mechanical, and histological properties of tibial bone. Month-old male Japanese quail were fed a basal diet containing either soybean oil (SBO), hydrogenated soybean oil (HSBO), chicken fat (CF), or menhaden fish oil (FO) at 50 g/kg of the diet and maintained on these diets for 7 mo. Lipid treatments did not affect body weight, food intake, tibial length, or diameter. The FO diet group had the highest percentage of tibial ash, and both the FO and HSBO significantly increased tibial mineral content compared to those given SBO or CF. The type and amount of fatty acids in the diets had a profound influence on fatty acid composition of lipids in tibial cortical bones. Quail fed FO had the highest concentration of (n-3) fatty acids, and those fed SBO were highest in (n-6) fatty acids. The HSBO diet, containing high level of trans-fatty acids, led to the accumulation of these fatty acids in bone. In quail, long-term supplementation of FO or HSBO increased tibial shear force and shear stress and improved histological cortical thickness and density when compared to those given SBO or CF. These results suggest that long-term exposure to a FO or HSBO diet have a significant beneficial effect on bone metabolism. PMID- 12762408 TI - Effects of storage time on incubating egg gas pressure, thyroid hormones, and corticosterone levels in embryos and on their hatching parameters. AB - Incubating eggs (1,800 total) produced by a commercial flock of Cobb broiler breeders were used to determine the effects of storage duration (3 and 18 d) on gas partial pressure, thyroid hormones, and hatching parameters. Partial pressure of oxygen (pO2) and carbon dioxide (pCO2) were measured on d 18 and at internal pipping (IP) during incubation. Blood samples were collected for determination of triiodothyronine (T3), thyroxine (T4), and corticosterone concentrations in the embryos at IP and in newly hatched chicks. From 464 to 510 h of incubation, eggs were checked individually every 2 h to determine the timing and duration of IP, external pipping (EP), and total hatching time. At 18 d of incubation and at IP, pCO2 was greater in air cell of eggs stored for 3 d compared to those stored for 18 d (P < 0.05), but pO2 was greater in eggs stored for 18 d. At IP, T3 and corticosterone levels were higher in plasma of the embryos of eggs stored for 3 d compared to those stored for 18 d, but it was the reverse in newly hatched chicks (P < 0.05). Embryos from eggs stored for 18 d required more time to complete IP compared to embryos of eggs stored for only 3 d (P < 0.05), whereas the duration of EP was not affected by storage. The overall longer incubation was, however, not only due to prolonged IP but also to later occurrence of IP. It was concluded that prolonged IP as a result of long storage may be related to the late increase in corticosterone level, which may be a necessary stimulus for higher T3/T4 ratio, late increase in pCO2 level, and decrease in pO2. The effect of long storage was a delay in hatching and a continuous increase in T3 due to higher corticosterone levels between IP and hatching, which may be an indication of the more stressful event of hatching of embryos from eggs stored longer. Differences in pCO2, pO2, T3, T4, and corticosterone levels in the incubating eggs may be manifestations of these changes culminating in altered hatching parameters and consequently differences in chick quality and growth potentials. PMID- 12762409 TI - Abdominal skin temperature variation in healthy broiler chickens as determined by thermography. AB - Abdominal skin temperature of healthy broiler chickens was determined by thermography to estimate the normal range of between- and within-bird temperature variation. Effects of potential confounding factors, such as bird's contention duration, abdominal side, age, and time of day, were also estimated. Mean skin temperature was estimated from thermogram pictures by computer image analysis of a predetermined abdominal area. Results demonstrated that skin temperature was not significantly affected by contention duration. However, skin temperature was higher on the right abdominal side in younger birds and at 1200 h. Between- and within-bird variation in healthy birds was small (< 0.5 degrees C) when age, abdominal side, and time of day were taken into account. This work suggests that skin temperature measurement using thermography can be adapted for the study of skin temperature in broiler chickens. PMID- 12762410 TI - Use of double packaging and antioxidant combinations to improve color, lipid oxidation, and volatiles of irradiated raw and cooked turkey breast patties. AB - The effects of antioxidants and double packaging combinations on color, lipid oxidation, and volatiles production in irradiated raw and cooked turkey breast were determined. Ground meat was treated with antioxidants (none, sesamol + alpha tocopherol, or gallate + alpha-tocopherol), and patties were prepared. The patties were packaged under vacuum, packaged aerobically, or double packaged (vacuum for 7 d then aerobic for 3 d) and electron beam irradiated at 3 kGy. Color, 2-thiobarbituric acid-reactive substances (TBARS), and volatile profiles of the samples were determined at 0 and 10 d and after cooking. Irradiated vacuum packaged patties had great amounts of sulfur volatiles (dimethyl sulfide and dimethyl disulfide) and increased red color during refrigerated storage and after cooking compared with the nonirradiated control. Irradiated aerobically packaged meat had accelerated lipid oxidation and aldehyde production at 10 d and after cooking. Gallate + alpha-tocopherol alone with double packaging was effective in reducing the red color of irradiated meat at 10 d and after cooking. Considerable amounts of off-odor volatiles were reduced by double packaging and antioxidant treatment. Sulfur volatiles were evaporated during the aerobic period of double packaging, and lipid oxidation was prevented by the antioxidants and vacuum condition of double packaging. These beneficial effects of double packaging and antioxidants were more critical in irradiated cooked meat. Therefore, the combined use of antioxidants and double packaging would be a useful method to control the oxidative quality changes of irradiated raw and cooked turkey breast. PMID- 12762411 TI - Impact of technical difficulties, choice of catheter, and the presence of blood on the success of embryo transfer--experience from a single provider. AB - PURPOSE: To investigate the impact of technical difficulties, choice of catheter, and the presence of blood during embryo transfer on the results of in vitro fertilization and ICSI. METHODS: A cohort of 784 consecutive cycles in 655 in vitro fertilization and ICSI patients were studied. RESULTS: Negotiating the cervix, using the volsellum, presence of blood on the catheter wall or on the cervix did not affect the results. Changing the catheter and blood on the catheter tip reduced the pregnancy (P < 0.05 and P < 0.05) and implantation rates (P < 0.001 and P < 0.01). The Ultrasoft catheter produced higher pregnancy (P < 0.0005) and implantation rates (P < 0.01) compared to the more rigid Frydman catheter. CONCLUSIONS: Negotiation of the cervix, the use of a volsellum, and the presence of blood on the catheter wall or on the cervix do not affect the results. Changing the catheter and blood on the catheter tip significantly diminish the pregnancy and implantation rates. Soft catheters perform better. PMID- 12762412 TI - Correlation between human follicular diameter and oocyte outcomes in an ICSI program. AB - PURPOSE: To determine the correlation between the follicular sizes and oocyte recovery, metaphase II oocyte recovery, fertilization rate and good embryo quality from mature and immature oocytes in an intracytoplasmic sperm injection (ICSI) program. METHODS: 991 follicles obtained from 72 ICSI cycles were classified into three groups according to their diameters as measured by transvaginal ultrasound including group A (< 10 mm), group B (10-14 mm), and group C (> 14 mm). All obtained oocytes were classified according to their nuclear maturation: germinal vesicle (GV), metaphase I (MI) and metaphase II (MII). Mature oocytes underwent ICSI while immature oocytes were further cultured until maturity before ICSI was performed. The rates of fertilization and good quality embryos at day 3 were evaluated. RESULTS: A progressive and significant increase in the rates of oocyte recovery and MII oocyte recovery were observed from group A follicles compared to the other groups (p < 0.001). The fertilization rate of mature and in vitro matured oocytes, as well as the rate of good quality embryos showed a tendency to increase from group A to group C follicles, but not significantly. The corresponding fertilization rates were 78 and 55.3% (p < 0.001) for mature and in vitro matured oocytes, respectively. CONCLUSION: Collection of oocytes from small follicles, especially with a mean diameter less than 10 mm, and in vitro maturation of immature oocytes before fertilization may allow the total number of good quality and transferable embryos to be increased. PMID- 12762413 TI - Development of murine embryos following electroporation. AB - PURPOSE: The objective was to establish the parameters for reversible electroporation of murine embryos. METHODS: In Trial 1, murine presumptive zygotes received an electrical pulse of 5, 10, or 20-micros duration, and one of five voltages (100, 200, 250, 300, or 400 V). In Trial 2, embryo orientation within the electroporation chamber was evaluated with 250 or 400 V at a pulse period of 10 micros. RESULTS: Presumptive zygotes that received 400 V at each pulse length and zygotes exposed to 20 micros at each voltage had the lowest embryonic development (P < 0.05). Presumptive zygotes that received 250 V had higher development compared to 400 V, irrespective of orientation (P < 0.01), but development was lower than the controls (P < 0.01). CONCLUSIONS: Electrical stimulation of presumptive zygotes can have a detrimental impact on early embryo development, but low amounts of stimulation may allow for potential gene transfer in transgenic experimentation. PMID- 12762414 TI - Rapid detection of the deltaF508 mutation in single cells using DHPLC: implications for preimplantation genetic diagnosis. AB - PURPOSE: Practice of preimplantation genetic diagnosis (PGD) requires efficient amplification and analysis techniques. We have tested Denaturing High Performance Liquid Chromatography (DHPLC) to screen the deltaF508 mutation in heterozygous single cells in order to assess its usefulness for PGD of cystic fibrosis. METHODS: One hundred and two single lymphocytes--including N/N and N/deltaF508- were studied. F508 locus was amplified by nested PCR followed by the analysis of PCR products by DHPLC in non-denaturing conditions. RESULTS: On the basis of PCR amplified product analysis, total efficiency of amplification was 98.78% (101/102), and allele dropout (ADO) rate was 3.7% (3/81). For each sample, results were obtained in less than 4 min with high resolution. CONCLUSIONS: DHPLC is a rapid and efficient technique to detect the deltaF508 mutation in single cells and is therefore appropriate for clinical application of preimplantation genetic diagnosis of cystic fibrosis. PMID- 12762415 TI - Does preejaculatory penile secretion originating from Cowper's gland contain sperm? AB - PURPOSE: To determine if spermatozoa are present in the preejaculatory penile secretion, originating from Cowper's gland. METHODS: DESIGN: Prospective clinical and laboratory study. SETTING: Andrology and Sex Counseling Unit, Department of Obstetrics and Gynecology, Academic Teaching Hospital. PATIENTS: Five patients referred for premature ejaculation, three for excessive fluid secreted during foreplay and four normal healthy volunteers. INTERVENTION: Glass slide smears of preejaculatory Cowper's gland secretion obtained during foreplay from at least two different occasions, and semen samples after masturbation. MAIN OUTCOME MEASURES: Microscopic examination of air-dried smears, and routine semen analyses. RESULTS: None of the preejaculatory samples contained sperm. All the patients had sperm in routine sperm analyses. CONCLUSIONS: Preejaculatory fluid secreted at the tip of the urethra from Cowper's gland during sexual stimulation did not contain sperm and therefore cannot be responsible for pregnancies during coitus interruptus. PMID- 12762416 TI - Isolation of ejaculated spermatozoa six hours after unsuccessful testicular biopsy in nonobstructive azoospermia. AB - Azoospermia, the absence of sperm in ejaculated semen, is the most severe form of male factor infertility and is present in approximately 5% of all investigated infertile couples. The condition is currently classified as obstructive and nonobstructive subgroups. In nonobstructive azoospermia, testicular sperm extraction (TESE) is usually necessary for sperm recovery. This is a case report of pregnancy and subsequent birth of healthy babies following intracytoplasmic sperm injection (ICSI) of ejaculated motile spermatozoa presented 6 h after unsuccessful testicular biopsies in four subsequent in vitro fertilization (IVF) cycles with infertility due to nonobstructive azoospermia. PMID- 12762417 TI - Towards finding the person in the data of personality. AB - This article is based on an address given on the occasion of receiving the 2000 Henry A. Murray award. The article presents a glimpse of my life story in personality and contributions to the field. These are placed in the context of observations about the recent history and sociology of the field. I outline some perspectives on the data that are collected and missing in personality research as well as the analyses that are conducted and those that are not conducted. These considerations identify both some persisting limitations in personality research and alternative analytic approaches that may prove useful in framing and answering new questions. Of particular promise are intensive studies that allow researchers to maintain a clear focus on the individual person. PMID- 12762419 TI - The Tripartite Model of Neuroticism and the suppression of depression and anxiety within an escalation of commitment dilemma. AB - We found evidence of a mutual suppression effect between anxiety and depression on an individual's level of commitment within escalation dilemmas. On the one hand, our results demonstrate a positive relationship between anxiety and level of commitment; on the other, our results demonstrate a negative relationship between depression and level of commitment. Based on the opposing relationships between anxiety and depression and commitment, the broad factor of neuroticism does not demonstrate any relationship with level of commitment, and the significant effects of anxiety and depression on commitment is contingent upon partialling the effect of the other facet of neuroticism. Thus, we contend that applied psychologists, who have focused on neuroticism as a broad construct, should consider the large body of work among clinical psychologists, who argue that anxiety and depression have unique variance associated with them. We conclude by addressing organizational implications of measuring the broad trait of neuroticism more narrowly. PMID- 12762418 TI - Sociability and positive emotionality: genetic and environmental contributions to the covariation between different facets of extraversion. AB - The relation between sociability and positive affect is one of the most often replicated results of research on personality and subjective well-being. It is shown how behavior genetics can contribute to our understanding of the covariance between sociability and positive emotionality. The results of a multimethod behavior-genetic study with 158 monozygotic and 120 dizygotic twins are reported. In this study, sociability and two components of positive emotionality (positive affect, energy) were assessed by self-report and other report. Additionally, positive state affect was assessed in five situations and aggregated across situations. The results showed that there are strong genetic correlations between all variables. Furthermore, there are substantive correlations between the nonshared environmental components of the different variables. Shared environmental influences, however, seemed to be unimportant for explaining the correlations between sociability and the different components of positive emotionality. The results are discussed with respect to their implications for future research on sociability and positive emotionality. PMID- 12762420 TI - A motivational analysis of defensive pessimism and self-handicapping. AB - Two studies examined motivational influences on and correlates of defensive pessimism and self-handicapping and investigated the relationship between these two cognitive strategies and performance attainment. The findings indicated that defensive pessimism and self-handicapping have similar motivational profiles, with the primary difference being that self-handicapping represents the absence of approach motivation in the achievement domain, as well as the presence of avoidance motivation. Self-handicapping, but not defensive pessimism, was shown to undermine performance-attainment, and performance-avoidance goals were validated as mediators of this negative relationship. Issues regarding the functional nature of the two cognitive strategies are discussed. PMID- 12762421 TI - A multilevel analysis of the interpersonal behavior of socially anxious people. AB - We investigate the interpersonal behavior of socially anxious (SA) and non socially anxious (NSA) individuals at three different levels of analysis, focusing on the dimensions of warmth and dominance. Study 1 examines self reported general interaction styles, Study 2 explores behavior occurring within the context of a single interaction, and in Study 3 we focus on the performance of a single conversational act (a disagreement). Studies 1 and 2 adopt the framework of Interpersonal Circumplex Theory (IPC; Kiesler, 1983), which is well suited for studying trait-level and interaction-level social behaviors, while Study 3 is grounded in Politeness Theory (PT; Brown & Levinson, 1987), which can be used to analyze individual acts at the microstructural level. The potential mutual relevance of PT and IPC is also discussed. PMID- 12762422 TI - Two spheres of belief in justice: extensive support for the bidimensional model of belief in a just world. AB - The purpose of this research comprising five studies (N = 666) was to further corroborate the bidimensional conceptualisation of the belief in a just world proposed by Lipkus et al. (1996). It was demonstrated that belief in a just world for the Self (and not for Others) was correlated to evaluations of the meaning of life. Belief in a just world for Others was significantly correlated to discrimination against the elderly, stigmatisation of poverty, and higher penal punitiveness, while belief in a just world for Self was weakly or not related to these variables. Together, these observations confirm the importance of the conceptual and psychometric distinction between these two spheres of the belief in a just world. PMID- 12762423 TI - Accurate intelligence assessments in social interactions: mediators and gender effects. AB - Research indicates that people can assess a stranger's measured intelligence more accurately than expected by chance, based on minimal information involving appearance and behavior. The present research documents behavioral correlates of perceived and measured intelligence and identifies behaviors that mediate the relationship between perceived and measured intelligence. In particular, when judges rated targets with video and auditory stimuli available, responsiveness to conversation partner, eye-gaze, and looking at partner while speaking were each significant mediators in the accurate assessment of intelligence. Each of those behaviors, as well as the percentage of looking at partner while speaking as a function of the target's own speaking time, were significant mediators in the video silent condition. Additionally, judge and target gender contributed to accurate intelligence assessments. PMID- 12762424 TI - Laboratory diagnosis of Hodgkin's disease: what are the challenges? PMID- 12762425 TI - Haematoxylin and eosin staining in the diagnosis of Hodgkin's disease in Uganda. AB - BACKGROUND: Whereas immunohistochemical methods have been widely used for the diagnosis and classification of Hodgkin's disease in the developed countries, there are very few reports of their use in the developing countries where haematoxylin and eosin is the mainstay of diagnosis of Hodgkin's disease. Yet the diagnostic accuracy of haematoxylin and eosin has not been assessed in Uganda. OBJECTIVE: To determine the reliability of haematoxylin and eosin staining in the diagnosis of Hodgkin's disease using immunohistochemistry as the reference standard. DESIGN: Laboratory based cross sectional study. SETTING: Makerere University Medical School, Department of Pathology. METHODOLOGY: Two hundred and forty formalin fixed, paraffin embedded biopsies seen in the Makerere University, Department of Pathology from 1980-2000 were studied. The tissue sections, were assessed and subjected to immunohistochemical methods using monoclonal antibodies including leucocyte common antigen, LCA (CD45), antibodies to Reed-Sternberg cells (CD15, CD30) and antibodies to B cells (CD20). The sensitivity, specificity, positive predictive value and negative predictive value were assessed. The overall Kappa score was used to assess the agreement between the two diagnostic tests. RESULTS: Of the 240 biopsies, 171(71.3%) were confirmed as Hodgkin's disease by immunohistochemistry. Using haematoxylin and eosin (H&E), only 131 of the 171 cases of Hodgkin's disease were detected. The mean age of the 171 cases was 26.1 (SD 16.2) years, with a mode of 20.0 and median of 22.5 years. The 15-24 year age group was the most affected (47.2%). There were more males (65.9%) than females and most were Baganda the dominant tribe in the central region. The sensitivity, specificity, positive and negative predictive values of haematoxylin and eosin were 76.61%, 92.75%, 96.32% and 61.53% respectively. The agreement between the two tests was 81.25% with an overall measure of agreement, Kappa, of 0.602. CONCLUSION: Haematoxylin and eosin has relatively high efficacy in the diagnosis of Hodgkin's disease. Use of haematoxylin and eosin is still recommended for the diagnosis of Hodgkin's disease, reserving the expensive immunohistochemistry for difficult cases. PMID- 12762426 TI - Topical phenytoin versus EUSOL in the treatment of non-malignant chronic leg ulcers. AB - OBJECTIVE: To compare topical diphenylhydantoin (phenytoin) with Edinburgh University solution of lime (EUSOL) in terms of rate of ulcer healing, analgesic and antibacterial properties in non-malignant chronic leg ulcers. DESIGN: A prospective randomized controlled study. SETTING: Muhimbili National Hospital surgical wards from August 2000 to September 2001. PATIENTS: One hundred and two patients with non-malignant chronic leg ulcers of various aetiologies, 50 in the study (phenytoin)group and 52 in the control(EUSOL) group. INTERVENTIONS: Study group studied by sprinkling phenytoin powder and the control group with EUSOL, in both groups the ulcers were addressed daily and followed up for 28 days or until they epithelialised or were ready for skin grafting. The data collected included demographic characteristics of patients, aetiology of the ulcers, pus discharge, severity of pain due to the ulcers, bacterial cultures from ulcer swabs, rate of reduction in mean ulcer surface area and outcome of treatment. RESULTS: The study enrolled 67 male and 35 female patients over a 14 month period (August to September 2001). Fifty patients formed the study group and 52 formed the controls. The age range was 12-56 years; the majority being in the 27-31 year age group. Major causes of chronic leg ulcers were those infected following trauma (27.5%), chronic non-specific inflammations (21.6%) and infected burn wounds (15.7%). At enrolment, the duration of ulcers ranged from 3-156 weeks and 3-128 weeks in the phenytoin and control groups respectively. Overall, there was significant reduction in pain(p < 0.05) on day seven in the phenytoin group. Furthermore in patients who presented with severe pain, there was a significant reduction in pain in the phenytoin group on the fourteenth day (p < 0.01). Clearance of ulcer discharge was also significant in the phenytoin group on the seventh and fourteenth day of treatment(p < 0.05). The commonest bacteria isolated were pseudomonas aeruginosa (54.9%) and staphylococcus aureus (10.8%). However, bacterial colonization clearance was not statistically significant when the two groups were compared. The rate of formation of healthy granulation tissue was highly significant in the phenytoin group by the fourteenth and twenty first days of treatment (p < 0.001). The phenytoin group showed significant reduction in the mean ulcer surface area on days 7, 14, 21 and 28 (p < 0.05). Chronic ulcers due to animal bites healed fastest followed by those due to trauma. CONCLUSION: Phenytoin powder is cheap and easily applied topically on ulcers, effectively relieves pain, clears discharge and enhances formation of granulation tissue thereby promoting healing; reducing morbidity and financial burden enabling its use in resource-poor environments. PMID- 12762427 TI - Chronic suppurative otitis media in school pupils in Nigeria. AB - OBJECTIVE: To compare the prevalence of chronic supperative otitis media (CSOM) among two populations of school children. DESIGN: A survey-demographic data collection and otoscopy was carried out among the two populations of school children. SETTING: A rural government primary school and an urban private primary school SUBJECTS: Six hundred and ninety nine school pupils in the rural school and two hundred and seventy pupils in the urban school. RESULTS: Six per cent of the pupils in the rural schools had CSOM as evidenced by persistent perforation of tympanic membrane of more than three months duration. No tympanic membrane perforations were observed in the children in the urban school at the time of this study. The difference in the prevalence of CSOM between the two populations is statistically significant (P < 0.001). The difference in socio-economic status between the two populations is statistically significant in relation to the prevalence of CSOM in the two populations. The poorer rural population had a significantly higher prevalence of CSOM (P < 0.001). CONCLUSION: The two study populations are just seven kilometres apart geographically but there is a world of difference in their socio-economic status, availability of social infrastructure and health facilities. This presumably had reflected in the marked difference in the prevalence of CSOM between the two study populations. A collective effort of government and well meaning indigenes of rural communities in Nigeria can help promote the socio-economic status and enhance the availability of social infrastructure and health facilities of rural areas. This we hope will lead to a decline in the prevalence of CSOM in the rural areas. PMID- 12762428 TI - Does preventive health care have a chance in the changing health sector in Tanzania? AB - OBJECTIVE: To investigate the status and practice of preventive health care (relative to curative) in the health delivery system at the time when the health sector reforms are taking place. DESIGN: A cross-sectional, descriptive study. SETTING: The study was conducted in Morogoro District between January and May 1999. SUBJECTS: Eighty six medical personnel and two hospital administrators from thirty four health facilities. The health facilities included twenty five dispensaries, five health centres and four hospitals. Care was also taken to include health facilities owned by various institutions and organisations, including governmental and non-governmental. RESULTS: Generally, preventive health received little attention compared to the curative health measures whereby more than 80% of the medical personnel in some of the facilities were assigned to curative services. Health personnel reported to spend an average of up to six hours per day providing curative services such as chemotherapy, surgical treatment, psychotherapy and radiography. On the contrary, they spent about four hours or less on providing child immunisation and education on nutrition, health and family planning. As expected, the type of ownership of a health facility influenced the extent to which preventive measures were included. For example, while all the government owned facilities did provide child immunisation, nutrition education and family planning services, some non-governmental facilities were lacking such services. CONCLUSION: It is obvious that while the provision of curative health care can be left to the hands of the private suppliers, that of preventive health care needs strong government involvement. It is suggested that deliberate efforts be taken to shift resources from curative to preventive measures. One way in which such a strategy can be attained is for the government to set, as a condition for private operators, a minimum level of preventive measures to be provided by every operator before a permit is issued. However, caution should be taken to ensure that such deliberations do not discourage investors in the health sector. PMID- 12762429 TI - Antenatal and intrapartum risk factors for birth asphyxia among emergency obstetric referrals in Mulago Hospital, Kampala, Uganda. AB - BACKGROUND: Many perinatal deaths follow birth asphyxia that occurs in newborn babies of women who are referred on developing life-threatening obstetric complications. OBJECTIVE: To determine the antenatal and intrapartum risk factors for severe birth asphyxia among babies delivered by women admitted as emergency obstetric referrals. DESIGN: Case-control study. SETTING: Mulago hospital, the National Referral Hospital, Kampala, Uganda. SUBJECTS: Cases were newborn term babies (and their mothers) with a 5-minute Apgar score 4 or less (birth asphyxia). Controls were term newborn babies with a 5-minute Apgar score more than 4. MAIN OUTCOME MEASURES: Antepartum and intrapartum risk factors among newborn babies (and their mothers) from socio-demographic characteristics, obstetric complications or labour management. The Odds ratios (OR) for various outcomes were calculated using the Statistical Assistance Software (SAS) version 6.2 (Windows), and are presented with their 95% confidence intervals (C1) and p values. RESULTS: There was no association between socio-demographic factors and birth asphyxia. Antepartum hospitalization, antepartum or intrapartum anaemia, antepartum hemorrhage and severe pre-eclampsia/eclampsia were significantly associated with birth asphyxia; the respective ORs and 95% C1 were 1.73 (1.09 2.75), 5.65 (3.36-9.50), 2.12 (1.11-4.05) and 10.62 (2.92-38.47). Augmentation of labour with oxytocin, premature rupture of membranes, meconium staining of liquor amnii, vacuum extraction, caesarean section, low birth weight and mal presentations were significantly associated with birth asphyxia with ORs of 5.76 (2.20-15.05), 2.23 (1.31 -3.37), 6.40 (2.76-14.82), 2.16 (1.28-3.67), 2.36 (1.07 5.20) and 6.32 (3.57-11.20) respectively. CONCLUSIONS: Early recognition of these complications among emergency obstetric referrals, followed by prompt and appropriate management, may reduce the perinatal deaths from birth asphyxia. PMID- 12762430 TI - Maternal mortality and associated near-misses among emergency intrapartum obstetric referrals in Mulago Hospital, Kampala, Uganda. AB - BACKGROUND: Many maternal deaths (as well as related severe morbidity) are of women who do not attend antenatal care in a given health unit but are referred there when they develop life-threatening obstetric complications. OBJECTIVE: To determine the reproductive characteristics of emergency obstetric referrals, and determine the contribution of emergency obstetric referrals to severe acute maternal morbidity (near-misses) and maternal mortality. STUDY DESIGN: Descriptive cross-sectional study. SETTING: Mulago hospital, the National Referral hospital, Kampala, Uganda, from 1st March to August 30th 2000. SUBJECTS: Nine hundred and eighty three consecutive women admitted as emergency obstetric referrals in labour or puerperium. INTERVENTIONS: Subjects were followed from time of admission to discharge (or death). They were interviewed (or examined) to obtain data on socio-demographic characteristics, reproductive history, obstetric outcome of the index pregnancy, obstetric complications and cause of death. Their records were reviewed to determine evidence of severe acute morbidity from acute organ/system dysfunction, using the definition by Mantel et al. These data were analysed using the Epilnfo computer programme in terms of means, frequencies and percentages. MAIN OUTCOME MEASURES: Socio-demographic characteristics, obstetric complications, cause of deaths, cause and type of near miss mortality and case fatality rates. RESULTS: Of the 983 referrals, over 100 were near-misses and 17 died. Using the definition of Mantel et al of near-misses enabled identification of six times as many near-misses as maternal deaths. The commonest causes of death were postpartum haemorrhage and eclampsia. Low status was highly associated with both maternal deaths and near misses. CONCLUSION: In developing countries, with poor obstetric services, emergency transfers in labour are very common. These women, who are of low status, contribute significantly to maternal mortality and morbidity. PMID- 12762431 TI - Does eradication of Helicobacter pylori reduce hypergastrinaemia during long term therapy with proton pump inhibitors? AB - OBJECTIVES: To evaluate the effect of Helicobacter pylori (Hp) eradication therapy on blood gastrin levels in long-term PPI users, since proton pump inhibitors (PPIs) and Helicobacter pylori (Hp) are major causes of hypergastrinaemia. DESIGN: A prospective study. SUBJECTS: Twenty seven Hp (+) patients enrolled in the study. Twenty were given eradication treatment (ET group), and the rest were given symptomatic treatment (ST group). Those who remained Hp (+) after eradication therapy were also added into the ST group. Lansoprazol 30 mg/day was given to both groups for three months thereafter. RESULTS: Fasting and non-fasting blood gastrin levels (FGL and NFGL) were measured initially and one month and four months after treatment. At the end of fourth month, FGL was significantly higher than both initial and first month level (p < 0.01) in the ST group. NFGL in this group did not change significantly (p > 0.05) after eradication therapy. In the ET group, FGL was significantly higher in the fourth month than the first month (p < 0.001) and than the initial level (p < 0.05). NFGL was higher, but not statistically in the fourth month than in the first month (p > 0.05) and significantly lower than the initial level (p < 0.05) in this group. CONCLUSION: We suggest that testing for Hp positivity and treating it if detected would be an appropriate approach to avoid hypergastrinaemia, especially in candidate patients for long term PPI treatment. PMID- 12762432 TI - Psychosocial correlates of substance use amongst secondary school students in south western Nigeria. AB - OBJECTIVE: To determine the psychosocial correlates of substance use among secondary school students in rural and urban communities in south western Nigeria. DESIGN: A cross-sectional survey of secondary school students using questionnaire eliciting substance use by students (WHO drug use questionnaire) and a well designed questionnaire on psychosocial variables. SETTING: Six secondary schools selected from two local government areas in Ilesa, Osun State, South Western Nigeria. SUBJECTS: The study population comprised 600 randomly selected senior secondary school students from six schools. RESULTS: A total of 562 questionnaires were analysed. Current stimulant use was significantly associated with lower socio-economic status, coming from a polygamous family and self-rated poor academic performance. Current alcohol use was associated with being a male, polygamous family background, living alone or with friends, not being religious and self-rated poor academic performance. Current hypnosedatives use was commoner in students living alone or with friends and in those with self rated poor academic performance. There was also significant positive relationship between current tobacco use and the male sex, not being religious and self-rated poor academic perfomance. Lifetime use of these substances had similar association with the psychosocial variables with slight differences. CONCLUSION: The similarity between the psychosocial correlates highlighted in this study and those reported in previous studies from other parts of Nigeria makes these observations useful enough for the planning of preventive strategies. PMID- 12762433 TI - Terrorist bombing at the USA Embassy in Nairobi: the media response. AB - BACKGROUND: On 7th August 1998, a terrorist bomb destroyed the American Embassy in Nairobi, killing and injuring thousands of people. OBJECTIVES: To describe the media component of the (larger) mental health response, and evaluate the impact of the intervention. DESIGN: Descriptive cross sectional study. SETTING: Nairobi, three months after the explosion. SUBJECTS: Residents of Nairobi and outlying areas, exposed to the bomb blast and who received Radio and TV coverage about the explosion. INTERVENTIONS: Radio and TV broadcasts, starting hours after the explosion and continuing for two weeks thereafter. RESULTS: Of a sample of 400 respondents interviewed, 172 reported they were affected by the bomb blast, either directly or through friends and relatives. TV and radio had the highest reach, (89% and 85%) respectively. Seventy per cent felt the programmes gave victims hope and 90% felt the programmes should continue, as they were helpful to the needy. CONCLUSION: The media is a controversial, sometimes overused and at times underused resource in Disaster Management. Judicious use of the media was shown to be helpful in this study. Literature supports media interventions in disaster. Doctors working in disaster areas need to understand the media and to prepare for its use as it compliments other interventions. PMID- 12762434 TI - Achalasia cardia and gastric outlet stenosis in a postmenopausal woman: case report. AB - The orderly contractility of the oesophagus and the regulated ability of the pyloric sphincter allow the influx and efflux of gastric contents. When these physiological processes are impaired, gastric luminal transit is altered as expected in achalasia cardia and gastric outlet obstruction. Movement across the inlet and outlet of the stomach is therefore altered. A case of a 58-year old woman diagnosed with simultaneous occurrence of achalasia cardia and gastric outlet stenosis resulting from chronic duodenal ulcer is presented. The diagnosis was based on clinical, radiological and intraoperative findings. This patient has remained well after a simultaneous anterior cardiomyotomy and H-M pyloroplasty. To my knowledge this is the first time that such an association causing gastric "inlet" and "outlet" obstruction has been reported. PMID- 12762435 TI - Are statistical contributions to medicine undervalued? AB - Econometricians Daniel McFadden and James Heckman won the 2000 Nobel Prize in economics for their work on discrete choice models and selection bias. Statisticians and epidemiologists have made similar contributions to medicine with their work on case-control studies, analysis of incomplete data, and causal inference. In spite of repeated nominations of such eminent figures as Bradford Hill and Richard Doll, however, the Nobel Prize in physiology and medicine has never been awarded for work in biostatistics or epidemiology. (The "exception who proves the rule" is Ronald Ross, who, in 1902, won the second medical Nobel for his discovery that the mosquito was the vector for malaria. Ross then went on to develop the mathematics of epidemic theory--which he considered his most important scientific contribution-and applied his insights to malaria control programs.) The low esteem accorded epidemiology and biostatistics in some medical circles, and increasingly among the public, correlates highly with the contradictory results from observational studies that are displayed so prominently in the lay press. In spite of its demonstrated efficacy in saving lives, the "black box" approach of risk factor epidemiology is not well respected. To correct these unfortunate perceptions, statisticians would do well to follow more closely their own teachings: conduct larger, fewer studies designed to test specific hypotheses, follow strict protocols for study design and analysis, better integrate statistical findings with those from the laboratory, and exercise greater caution in promoting apparently positive results. PMID- 12762436 TI - Sensitivity analyses for ecological regression. AB - In many ecological regression studies investigating associations between environmental exposures and health outcomes, the observed relative risks are in the range 1.0-2.0. The interpretation of such small relative risks is difficult due to a variety of biases--some of which are unique to ecological data, since they arise from within-area variability in exposures/confounders. The potential for residual spatial dependence, due to unmeasured confounders and/or data anomalies with spatial structure, must also be considered, though it often will be of secondary importance when compared to the likely effects of unmeasured confounding and within-area variability in exposures/confounders. Methods for addressing sensitivity to these issues are described, along with an approach for assessing the implications of spatial dependence. An ecological study of the association between myocardial infarction and magnesium is critically reevaluated to determine potential sources of bias. It is argued that the sophistication of the statistical analysis should not outweigh the quality of the data, and that finessing models for spatial dependence will often not be merited in the context of ecological regression. PMID- 12762437 TI - Likelihood inference for exchangeable binary data with varying cluster sizes. AB - This article investigates maximum likelihood estimation with saturated and unsaturated models for correlated exchangeable binary data, when a sample of independent clusters of varying sizes is available. We discuss various parameterizations of these models, and propose using the EM algorithm to obtain maximum likelihood estimates. The methodology is illustrated by applications to a study of familial disease aggregation and to the design of a proposed group randomized cancer prevention trial. PMID- 12762438 TI - Testing the correlation for clustered categorical and censored discrete time-to event data when covariates are measured without/with errors. AB - In the analysis of clustered categorical data, it is of common interest to test for the correlation within clusters, and the heterogeneity across different clusters. We address this problem by proposing a class of score tests for the null hypothesis that the variance components are zero in random effects models, for clustered nominal and ordinal categorical responses. We extend the results to accommodate clustered censored discrete time-to-event data. We next consider such tests in the situation where covariates are measured with errors. We propose using the SIMEX method to construct the score tests for the null hypothesis that the variance components are zero. Key advantages of the proposed score tests are that they can be easily implemented by fitting standard polytomous regression models and discrete failure time models, and that they are robust in the sense that no assumptions need to be made regarding the distributions of the random effects and the unobserved covariates. The asymptotic properties of the proposed tests are studied. We illustrate these tests by analyzing two data sets and evaluate their performance with simulations. PMID- 12762439 TI - Marginal analyses of clustered data when cluster size is informative. AB - We propose a new approach to fitting marginal models to clustered data when cluster size is informative. This approach uses a generalized estimating equation (GEE) that is weighted inversely with the cluster size. We show that our approach is asymptotically equivalent to within-cluster resampling (Hoffman, Sen, and Weinberg, 2001, Biometrika 73, 13-22), a computationally intensive approach in which replicate data sets containing a randomly selected observation from each cluster are analyzed, and the resulting estimates averaged. Using simulated data and an example involving dental health, we show the superior performance of our approach compared to unweighted GEE, the equivalence of our approach with WCR for large sample sizes, and the superior performance of our approach compared with WCR when sample sizes are small. PMID- 12762440 TI - A proposal for a goodness-of-fit test to the Arnason-Schwarz multisite capture recapture model. AB - In an analysis of capture-recapture data, the identification of a model that fits is a critical step. For the multisite (also called multistate) models used to analyze data gathered at several sites, no reliable test for assessing fit is currently available. We propose a test for the JMV model, a simple generalization of the Arnason-Schwarz (AS) model, in the form of interpretable contingency tables. For the AS model, we suggest complementing the test for the JMV model with a likelihood ratio test of AS vs. JMV. The examination of an example leads us to propose further a partitioning that emphasizes the role of the memory model of Brownie et al. (1993 Biometrics 49, 1173-1187) as a biologically more plausible alternative to the AS model. PMID- 12762441 TI - The analysis of ring-recovery data using random effects. AB - We show how random terms, describing both yearly variation and overdispersion, can easily be incorporated into models for mark-recovery data, through the use of Bayesian methods. For recovery data on lapwings, we show that the incorporation of the random terms greatly improves the goodness of fit. Omitting the random terms can lead to overestimation of the significance of weather on survival, and overoptimistic prediction intervals in simulations of future population behavior. Random effects models provide a natural way of modeling overdispersion-which is more satisfactory than the standard classical approach of scaling up all standard errors by a uniform inflation factor. We compare models by means of Bayesian p values and the deviance information criterion (DIC). PMID- 12762442 TI - Bayesian meta-analysis for longitudinal data models using multivariate mixture priors. AB - We propose a class of longitudinal data models with random effects that generalizes currently used models in two important ways. First, the random effects model is a flexible mixture of multivariate normals, accommodating population heterogeneity, outliers, and nonlinearity in the regression on subject specific covariates. Second, the model includes a hierarchical extension to allow for meta-analysis over related studies. The random-effects distributions are decomposed into one part that is common across all related studies (common measure), and one part that is specific to each study and that captures the variability intrinsic between patients within the same study. Both the common measure and the study-specific measures are parameterized as mixture-of-normals models. We carry out inference using reversible jump posterior simulation to allow a random number of terms in the mixtures. The sampler takes advantage of the small number of entertained models. The motivating application is the analysis of two studies carried out by the Cancer and Leukemia Group B (CALGB). In both studies, we record for each patient white blood cell counts (WBC) over time to characterize the toxic effects of treatment. The WBCs are modeled through a nonlinear hierarchical model that gathers the information from both studies. PMID- 12762443 TI - Bayesian multiple testing for two-sample multivariate endpoints. AB - In clinical studies involving multiple variables, simultaneous tests are often considered where both the outcomes and hypotheses are correlated. This article proposes a multivariate mixture prior on treatment effects, that allows positive probability of zero effect for each hypothesis, correlations among effect sizes, correlations among binary outcomes of zero versus nonzero effect, and correlations among the observed test statistics (conditional on the effects). We develop a Bayesian multiple testing procedure, for the multivariate two-sample situation with unknown covariance structure, and obtain the posterior probabilities of no difference between treatment regimens for specific variables. Prior selection methods and robustness issues are discussed in the context of a clinical example. PMID- 12762444 TI - Bayesian modeling of time-varying and waning exposure effects. AB - In epidemiologic studies, there is often interest in assessing the association between exposure history and disease incidence. For many diseases, incidence may depend not only on cumulative exposure, but also on the ages at which exposure occurred. This article proposes a flexible Bayesian approach for modeling age varying and waning exposure effects. The Cox model is generalized to allow the hazard of disease to depend on an integral, across the exposed ages, of a piecewise polynomial function of age, multiplied by an exponential decay term. Linearity properties of the model facilitate posterior computation via a Gibbs sampler, which generalizes previous algorithms for Cox regression with time dependent covariates. The approach is illustrated by an application to the study of protective effects of breastfeeding on incidence of childhood asthma. PMID- 12762445 TI - Generalized conjugate priors for Bayesian analysis of risk and survival regressions. AB - Conjugate priors for Bayesian analyses of relative risks can be quite restrictive, because their shape depends on their location. By introducing a separate location parameter, however, these priors generalize to allow modeling of a broad range of prior opinions, while still preserving the computational simplicity of conjugate analyses. The present article illustrates the resulting generalized conjugate analyses using examples from case-control studies of the association of residential wire codes and magnetic fields with childhood leukemia. PMID- 12762446 TI - A causal proportional hazards estimator for the effect of treatment actually received in a randomized trial with all-or-nothing compliance. AB - Survival data from randomized trials are most often analyzed in a proportional hazards (PH) framework that follows the intention-to-treat (ITT) principle. When not all the patients on the experimental arm actually receive the assigned treatment, the ITT-estimator mixes its effect on treatment compliers with its absence of effect on noncompliers. The structural accelerated failure time (SAFT) models of Robins and Tsiatis are designed to consistently estimate causal effects on the treated, without direct assumptions about the compliance selection mechanism. The traditional PH-model, however, has not yet led to such causal interpretation. In this article, we examine a PH-model of treatment effect on the treated subgroup. While potential treatment compliance is unobserved in the control arm, we derive an estimating equation for the Compliers PROPortional Hazards Effect of Treatment (C-PROPHET). The jackknife is used for bias correction and variance estimation. The method is applied to data from a recently finished clinical trial in cancer patients with liver metastases. PMID- 12762447 TI - Sequential tests for noninferiority and superiority. AB - The problem of simultaneous sequential tests for noninferiority and superiority of a treatment, as compared to an active control, is considered in terms of continuous hierarchical families of one-sided null hypotheses, in the framework of group sequential and adaptive two-stage designs. The crucial point is that the decision boundaries for the individual null hypotheses may vary over the parameter space. This allows one to construct designs where, e.g., a rigid stopping criterion is chosen, rejecting or accepting all individual null hypotheses simultaneously. Another possibility is to use monitoring type stopping boundaries, which leave some flexibility to the experimenter: he can decide, at the interim analysis, whether he is satisfied with the noninferiority margin achieved at this stage, or wants to go for more at the second stage. In the case where he proceeds to the second stage, he may perform midtrial design modifications (e.g., reassess the sample size). The proposed approach allows one to "spend," e.g., less of alpha for an early proof of noninferiority than for an early proof of superiority, and is illustrated by typical examples. PMID- 12762448 TI - A case-control follow-up study for disease-specific mortality. AB - Case-control studies often rely on subjects to report their own screening or exposure information: this information is often obtained from cases after the event of interest has occurred. This is problematic for mortality outcomes, because dead subjects cannot report the desired information. To avoid this problem, Weiss and Lazovich (1996, American Journal of Epidemiology 143, 319-322) proposed obtaining exposure or screening information from potential cases, i.e., subjects diagnosed with disease, at the time of disease diagnosis, and also from a referent series. The design is best viewed as a new scheme for sampling from a cohort. I review estimation of the effects of time-varying screening or exposure in cohort studies, using a new factorization. I then show how this factorization, together with ignorability assumptions, allows valid estimation from these new designs. Even when the sampling fraction of nondiseased subjects is unknown, causal risk ratios are estimable if diagnosis is rare in the cohort. I illustrate and compare conventional and new methods with data from the Health Insurance Plan study. PMID- 12762449 TI - Penalized estimating equations. AB - Penalty models--such as the ridge estimator, the Stein estimator, the bridge estimator, and the Lasso-have been proposed to deal with collinearity in regressions. The Lasso, for instance, has been applied to linear models, logistic regressions, Cox proportional hazard models, and neural networks. This article considers the bridge penalty model with penalty sigma(j)/beta(j)/gamma for estimating equations in general and applies this penalty model to the generalized estimating equations (GEE) in longitudinal studies. The lack of joint likelihood in the GEE is overcome by the penalized estimating equations, in which no joint likelihood is required. The asymptotic results for the penalty estimator are provided. It is demonstrated, with a simulation and an application, that the penalized GEE potentially improves the performance of the GEE estimator, and enjoys the same properties as linear penalty models. PMID- 12762450 TI - Selecting differentially expressed genes from microarray experiments. AB - High throughput technologies, such as gene expression arrays and protein mass spectrometry, allow one to simultaneously evaluate thousands of potential biomarkers that could distinguish different tissue types. Of particular interest here is distinguishing between cancerous and normal organ tissues. We consider statistical methods to rank genes (or proteins) in regards to differential expression between tissues. Various statistical measures are considered, and we argue that two measures related to the Receiver Operating Characteristic Curve are particularly suitable for this purpose. We also propose that sampling variability in the gene rankings be quantified, and suggest using the "selection probability function," the probability distribution of rankings for each gene. This is estimated via the bootstrap. A real dataset, derived from gene expression arrays of 23 normal and 30 ovarian cancer tissues, is analyzed. Simulation studies are also used to assess the relative performance of different statistical gene ranking measures and our quantification of sampling variability. Our approach leads naturally to a procedure for sample-size calculations, appropriate for exploratory studies that seek to identify differentially expressed genes. PMID- 12762451 TI - Data reduction using a discrete wavelet transform in discriminant analysis of very high dimensionality data. AB - We present a method of data reduction using a wavelet transform in discriminant analysis when the number of variables is much greater than the number of observations. The method is illustrated with a prostate cancer study, where the sample size is 248, and the number of variables is 48,538 (generated using the ProteinChip technology). Using a discrete wavelet transform, the 48,538 data points are represented by 1271 wavelet coefficients. Information criteria identified 11 of the 1271 wavelet coefficients with the highest discriminatory power. The linear classifier with the 11 wavelet coefficients detected prostate cancer in a separate test set with a sensitivity of 97% and specificity of 100%. PMID- 12762452 TI - Estimating causal treatment effects from longitudinal HIV natural history studies using marginal structural models. AB - Several recently completed and ongoing studies of the natural history of HIV infection have generated a wealth of information about its clinical progression and how this progression is altered by therepeutic interventions and environmental factors. Natural history studies typically follow prospective cohort designs, and enroll large numbers of participants for long-term prospective follow-up (up to several years). Using data from the HIV Epidemiology Research Study (HERS), a six-year natural history study that enrolled 871 HIV infected women starting in 1993, we investigate the therapeutic effect of highly active antiretroviral therapy regimens (HAART) on CD4 cell count using the marginal structural modeling framework and associated estimation procedures based on inverse-probability weighting (developed by Robins and colleagues). To evaluate treatment effects from a natural history study, specialized methods are needed because treatments are not randomly prescribed and, in particular, the treatment-response relationship can be confounded by variables that are time varying. Our analysis uses CD4 data on all follow-up visits over a two-year period, and includes sensitivity analyses to investigate potential biases attributable to unmeasured confounding. Strategies for selecting ranges of a sensitivity parameter are given, as are intervals for treatment effect that reflect uncertainty attributable both to sampling and to lack of knowledge about the nature and existence of unmeasured confounding. To our knowledge, this is the first use in "real data" of Robins's sensitivity analysis for unmeasured confounding (Robins, 1999a, Synthese 121, 151-179). The findings from our analysis are consistent with recent treatment guidelines set by the U.S. Panel of the International AIDS Society (Carpenter et al., 2000, Journal of the American Medical Association 280, 381-391). PMID- 12762453 TI - Accounting for nonignorable verification bias in assessment of diagnostic tests. AB - A "gold" standard test, providing definitive verification of disease status, may be quite invasive or expensive. Current technological advances provide less invasive, or less expensive, diagnostic tests. Ideally, a diagnostic test is evaluated by comparing it with a definitive gold standard test. However, the decision to perform the gold standard test to establish the presence or absence of disease is often influenced by the results of the diagnostic test, along with other measured, or not measured, risk factors. If only data from patients who received the gold standard test were used to assess the test performance, the commonly used measures of diagnostic test performance--sensitivity and specificity--are likely to be biased. Sensitivity would often be higher, and specificity would be lower, than the true values. This bias is called verification bias. Without adjustment for verification bias, one may possibly introduce into the medical practice a diagnostic test with apparent, but not truly, high sensitivity. In this article, verification bias is treated as a missing covariate problem. We propose a flexible modeling and computational framework for evaluating the performance of a diagnostic test, with adjustment for nonignorable verification bias. The presented computational method can be utilized with any software that can repetitively use a logistic regression module. The approach is likelihood-based, and allows use of categorical or continuous covariates. An explicit formula for the observed information matrix is presented, so that one can easily compute standard errors of estimated parameters. The methodology is illustrated with a cardiology data example. We perform a sensitivity analysis of the dependency of verification selection process on disease. PMID- 12762454 TI - Bootstrap choice of estimators in parametric and semiparametric families: an extension of EIC. AB - Ishiguro, Sakamoto, and Kitagawa (1997, Annals of the Institute of Statistical Mathematics 49, 411-434) proposed EIC as an extension of Akaike criterion (AIC); the idea leading to EIC is to correct the bias of the log-likelihood, considered as an estimator of the Kullback-Leibler information, using bootstrap. We develop this criterion for its use in multivariate semiparametric situations, and argue that it can be used for choosing among parametric and semiparametric estimators. A simulation study based on aregression model shows that EIC is better than its competitors although likelihood cross-validation performs nearly as well except for small sample size. Its use is illustrated by estimating the mean evolution of viral RNA levels in a group of infants infected by HIV. PMID- 12762455 TI - Modeling breastmilk infectivity in HIV-1 infected mothers. AB - Estimation of breastmilk infectivity in HIV-1 infected mothers is difficult because transmission can occur while the fetus is in utero, during delivery, or through breastfeeding. Since transmission can only be detected through periodic testing, however, it may be impossible to determine the actual mode of transmission in any individual child. In this article we develop a model to estimate breastmilk infectivity, along with the probabilities of in-utero and intrapartum transmission. In addition, the model allows separate estimation of early and late breastmilk infectivity, and individual variation in maternal infectivity. Methods for hypothesis testing of binary risk factors and a method for assessing goodness of fit are also described. Data from a randomized trial of breastfeeding versus formula feeding among HIV-1 infected mothers in Nairobi, Kenya, are used to illustrate the methods. PMID- 12762456 TI - Comment on Cowling's "Spatial methods for line transect surveys". AB - We consider the problem of estimating the parameters of a two-dimensional Neyman Scott process, from data collected through a line transect survey. Cowling (1998, Biometrics 54, 828-839) suggested an estimation method based on a one-dimensional K-function along the transect line. However, her expression for the theoretical K function is wrong. In this article, we correct her K-function. PMID- 12762457 TI - Latent class model diagnosis from a frequentist point of view. AB - This is in response to Garrett and Zeger (2000, Biometrics 56, 1055-1067) who, within the Bayesian framework, developed mainly graphical methods for latent class model diagnosis. Possible problems with this approach, and with its application to both generated and empirical data, are pointed out. The impact of the proposed tools cannot be understood by their reader, as no comparisons are made to results obtainable using established methods for latent class model diagnosis; this applies especially to overall goodness-of-fit tests, for which alternatives (bootstrap, Rudas-Clogg-Lindsay index of fit) are mentioned. Further, in one case of generated data, the methods proposed by Garrett and Zeger seem to give problematic results as to identifiability; in the case of the empirical data on major depression, they lead to accepting a suboptimal three class model. In the latter case, one can be rather sure that an identifiable, well-fitting latent class model could have been identified--if Garrett and Zeger had also considered restricted latent class models. PMID- 12762458 TI - Medical technology, end-of-life care and nursing ethics. PMID- 12762459 TI - Ethical issues regarding human cloning: a nursing perspective. AB - Advances in cloning technology and successful cloning experiments in animals raised concerns about the possibility of human cloning in recent years. Despite many objections, this is not only a possibility but also a reality. Human cloning is a scientific revolution. However, it also introduces the potential for physical and psychosocial harm to human beings. From this point of view, it raises profound ethical, social and health related concerns. Human cloning would have an impact on the practice of nursing because it could result in the creation of new physiological and psychosocial conditions that would require nursing care. The nursing profession must therefore evaluate the ethics of human cloning, in particular the potential role of nurses. This article reviews the ethical considerations of reproductive human cloning, discusses the main reasons for concern, and reflects a nursing perspective regarding this issue. PMID- 12762460 TI - Safeguarding being: a bioethical principle for genetic nursing care. AB - This philosophical inquiry examines the nature of the technology of genetic predisposition testing and its relation to patients as whole persons. The bioethical principles of non-maleficence, beneficence, autonomy and justice are judged insufficient to resolve issues associated with use. A new principle of 'sustained being', drawn from philosophical propositions of Pellegrino, is suggested. The new principle is suited to an evolving practice and is compatible with consequentialist, deontological and relational ethics theories. The notion of 'taking care' is related to nursing in genetic health care to form a standard of conduct and a moral imperative to 'safeguard being'. PMID- 12762461 TI - Do nurses exercise power in basic care situations? AB - Power is a matter of authority and control. It can be wielded either consciously or unconsciously, and it can be either overt or latent. Using a structured questionnaire, this study set out to describe nurses' opinions about the exercise of power in basic care situations in both acute and long-term care. The questionnaire was organized into four categories in which items concerned: power in obligatory daily activities; power in activities necessitated by obligatory activities; power in voluntary activities; and power in activities that take into account the patient's characteristics. The samples consisted of 228 nurses from five medical and surgical wards of district hospitals, and 233 nurses from five geriatric units of a community health centre and from one nursing home in Finland. The final response rate was 65% (acute care 76%; long-term care 55%). Data analysis was based on statistical methods. The results showed that, in the nurses' own opinion, negative power is exercised only in certain situations and in the patient's best interest, when for instance there are concerns that something may happen to the patient. PMID- 12762462 TI - Ethical dilemmas related to the HIV-positive person in the workplace. AB - This study's objectives were: (1) to describe and explore the ethical dilemmas surrounding the HIV-positive person in the workplace in South Africa; and (2) to describe the Rational Interaction for Moral Sensitivity (RIMS) approach as a possible mechanism for solving these ethical dilemmas. A qualitative, exploratory and descriptive research design was used. The target populations were HIV positive employees and occupational health nurses working for a South African company. Data collected through individual HIV-positive employee interviews and occupational health nurse workgroups were analysed. The ethical dilemmas were conceptualized and described within the theoretical framework of the principles of ethics, namely, autonomy, beneficence, justice and confidentiality. To elicit a solution to the dilemmas, the data were recontextualized using the RIMS approach, a group decision-making strategy designed for the business environment. PMID- 12762463 TI - A comparative study of Chinese, American and Japanese nurses' perceptions of ethical role responsibilities. AB - This article reports a survey of nurses in different cultural settings to reveal their perceptions of ethical role responsibilities relevant to nursing practice. Drawing on the Confucian theory of ethics, the first section attempts to understand nursing ethics in the context of multiple role relationships. The second section reports the administration of the Role Responsibilities Questionnaire (RRQ) to a sample of nurses in China (n = 413), the USA (n = 163), and Japan (n = 667). Multidimensional preference analysis revealed the patterns of rankings given by the nurses to the statements they considered as important ethical responsibilities. The Chinese nurses were more virtue based in their perception of ethical responsibilities, the American nurses were more principle based, and the Japanese nurses were more care based. The findings indicate that the RRQ is a sensitive instrument for outlining the embedded sociocultural factors that influence nurses' perceptions of ethical responsibilities in the realities of nursing practice. This study could be important in the fostering of partnerships in international nursing ethics. PMID- 12762464 TI - Moral stress: synthesis of a concept. AB - The aim of this article is to describe the synthesis of the concept of moral stress and to attempt to identify its preconditions. Qualitative data from two independent studies on professional issues in nursing were analysed from a hypothetical-deductive approach. The findings indicate that moral stress is independent of context-given specific preconditions: (1) nurses are morally sensitive to the patient's vulnerability; (2) nurses experience external factors preventing them from doing what is best for the patient; and (3) nurses feel that they have no control over the specific situation. The findings from this analysis are supported by recent research on stress in the workplace but differ that the imperatives directing work are moral in nature. Stress researchers have found that persons who experience that they have no control over their work situation and at the same time experience high demands may be prone to cardiovascular diseases. An important question raised by this study is whether moral stress should be recognized as a health risk in nursing. Further research is required in order to generate intervention models to prevent or deal with moral stress. PMID- 12762466 TI - Changes in ethical aspects of nursing. Interview by Arie van der Arend. PMID- 12762467 TI - Informed consent--'NURSE'. PMID- 12762468 TI - Remembering No-England. PMID- 12762469 TI - The "golden age" of dentistry: share the wealth. PMID- 12762470 TI - Dentin bonding. PMID- 12762471 TI - Esthetic soft tissue ridge augmentation using an acellular dermal connective tissue allograft. AB - Residual ridge defects present significant limitations to the esthetic restoration of edentulous tooth sites. Augmentation techniques can aid in the re establishment of natural ridge contours. This article describes a technique in which an acellular dermal connective tissue allograft is used to augment a buccolingual residual ridge defect. A single vertical incision enables the allograft to be placed laterally into a buccal pouch. Augmentation of this edentulous site permitted a three-unit bridge to be placed with a highly esthetic pontic. CLINICAL SIGNIFICANCE: This article presents a simplified method of esthetic ridge augmentation that reduces patient discomfort. PMID- 12762472 TI - Transient root resorption after dental trauma: the clinician's dilemma. AB - The diagnostic shortcomings encountered in the clinical evaluation of pulpal and periodontal healing subsequent to trauma comprise a major problem in dental traumatology. An acute dental trauma may imply impact to the hard dental tissues and damage to the pulp and periodontium (including surrounding alveolar bone). In the case of luxation injuries, the trauma often results in rupture of the neurovascular supply at the level of the apical foramen, whereas in a root fracture, the same occurs at the level of the fracture. Conventional diagnostic criteria for pulpal status (coronal discoloration, loss of pulpal sensibility, tenderness to percussion, and radiographic change [i.e., resorption]) reflect, only indirectly, certain parameters of healing. These parameters are based empirically on clinical findings after dental caries and dental restorative techniques, where the pulp has an intact neurovascular supply apically. In these situations, long-standing bacterial or chemical attack can lead to irreversible change. In contrast, following traumatic impact, a presumably intact pulp is deprived of its neurovascular supply instantaneously. In the absence of infection, events following acute dental trauma aim at either tissue repair or regeneration. Thus, based on a series of long-term clinical studies, it was found that all conventional signs, with the exception of tenderness to percussion, could be signs of pulpal healing, as well as signs of pulpal death. Based on the results of these studies, this report discusses the significance of radiographic change (resorption) in relation to pulpal and periodontal healing following acute trauma. Moreover, implications of tooth luxation with respect to the long-term prognosis following crown, crown-root, and root fractures are discussed. CLINICAL SIGNIFICANCE: Whenever possible, it is to the patient's advantage that pulp vitality be preserved. Correct diagnostic techniques and patient selection enable the clinician to decide when endodontic therapy is appropriate and when intelligent observation is the "treatment" of choice. PMID- 12762474 TI - Efficacy of a resin coating on bond strengths of resin cement to dentin. AB - PURPOSE: The aims of this study were to (1) evaluate the effect of a resin coating consisting of a dentin bonding system and a flowable resin composite on the microtensile bond strength (micro-TBS) of a resin cement to dentin in indirect composite restorations and (2) compare the bond strengths of direct and indirect composite restorations. MATERIALS AND METHOD: Occlusal surfaces of human premolars were ground to obtain flat dentin surfaces and were divided into seven groups. For indirect restorations, the dentin surfaces of the experimental groups were bonded with a dentin bonding system (DBS), Clearfil SE Bond (SE) or Single Bond (SB) with and without a flowable resin composite, Protect Liner F (PLF), temporized for one day and cemented with a resin cement (Panavia F) according to the manufacturer's instructions. The dentin surfaces of the control group were temporized without prior treatment, and indirect composite (Estenia) was bonded with Panavia F. For the direct restorations, either SE or SB was applied to the dentin surface and the entire surface was built up with direct composite (Clearfil AP-X). After 24 hours in water storage, micro-TBS was measured at a crosshead speed of 1 mm/min. The data were analyzed with one-way analysis of variance and Fisher's protected least significant difference test (p < .05). RESULTS: The original bond strength of the resin cement (Panavia F) to dentin significantly improved with the use of a resin coating technique in indirect restorations (p < .05). The combination of DBS + PLF showed significantly higher bond strengths compared with the single use of DBS. The combination of SE + PLF as a resin coating provided the highest bond strengths in indirect restorations (p < .05). However, the best bond strengths were observed when SE and SB were used for direct composite restorations (p < .05). CONCLUSIONS: The application of a resin coating consisting of a dentin bonding system and a flowable resin composite on the dentin following cavity preparation significantly improved the micro-TBS of the resin cement Panavia F to dentin in indirect restorations. However, the bond strengths of indirect composite restorations were significantly lower than those of direct composite restorations even with the resin coating technique. CLINICAL SIGNIFICANCE: Indirect composite restorations may require a resin coating to improve the bonding performance of the resin cement Panavia F to dentin. However, adhesive systems with direct composite restorations still provide superior bond strengths compared with indirect restorations. PMID- 12762473 TI - Reduction of polymerization shrinkage stress and marginal microleakage using soft start polymerization. AB - PURPOSE: This study evaluated the influence of a soft-start light-curing exposure on polymerization shrinkage stress and marginal integrity of adhesive restorations. MATERIALS AND METHODS: Six resin-based composites (Pertac II, Tetric Ceram, Definite, Surefil, Solitaire, and Visio-Molar) were adhesively bonded to a cylindrical cavity (n = 9 per material/light) in a photoelastic material. Visible light-curing was applied using either the standard polymerization mode (800 mW/cm2 exposure duration 40 s) of the curing light (Elipar TriLight, 3M ESPE) or the exponential mode from the same device (ramp curing: 150 mW/cm2 to 800 mW/cm2 within the first 15 s of a total curing time of 40 s). Polymerization stress was calculated at 5 minutes, 1 hour, and 24 hours postexposure from the second-order isochromatic curves obtained from photoelastic images (Matrox-Inspector). Two standardized Class V preparations were made each on the facial and lingual surfaces of 80 extracted human molars and premolars. Resin restorative systems (Pertac II/EBS Multi, Tetric Ceram/Syntac, Definite/Etch&Prime 3.0, and Surefil/Prime & Bond 2.1) were exposed using both light exposure modes (n = 20). Marginal dye penetration (2% methylene blue) was investigated separately for enamel and cementum margins after thermocycling. To obtain information on equivalent depth of cure, relative surface hardness measurements were performed on resin samples of the same material at the top surface and at 1.5 mm and 3.0 mm thickness (Zwick 3212, 10 N). RESULTS: A significant (p < .01) reduction in polymerization stress of 7.1% for Pertac II, 4.1% for Tetric Ceram, 3.6% for Definite, 3.7% for Surefil, and 6.2% for Solitaire was observed when using the exponential mode as opposed to the standard. A significant (p = .04) reduction of marginal dye penetration was found only for Pertac II/EBS Multi at the cementum margins when the soft-start polymerization was used. For the sample thickness of 3 mm, a significant higher relative bottom to top surface ratio in favor of the standard exposure mode was found (p = .001). CLINICAL SIGNIFICANCE: Depending on the restorative material, soft-start polymerization may lead to a significant reduction in marginal microleakage of adhesive Class V restorations. This effect might be attributable to a significantly lower polymerization stress, as seen from photoelastic analysis, and/or a decrease in the degree of conversion, as deducted from surface hardness ratios. However, the effect of soft-start curing mode depends on the material itself, with the most effective response from hybrid resin-based composites. PMID- 12762476 TI - Light-emitting diode curing lights, Part I. PMID- 12762475 TI - Color measurements as quality criteria for clinical shade matching of porcelain crowns. AB - STATEMENT OF PROBLEM: The ability of a dentist to select and communicate an acceptable shade match to a dental laboratory may be the most important factor in esthetic restorative dentistry. PURPOSE: The purpose of this study was to evaluate the use of instrumental color measurement in clinical shade matching of porcelain-fused-to-metal (PFM) and all-porcelain crowns. The relative effects of clinical and laboratory factors related to shade matching for PFM and all porcelain crowns were also evaluated. MATERIALS AND METHOD: Forty patients treatment planned to receive PFM or all-porcelain crowns made up the study population. The patients were randomly divided into two groups for shade selection: conventional visual assessment and photocolorimetric analysis. At the preparation appointment, a photograph was taken of the target tooth along with four shade guide tabs selected by the two visual observers. The crown was fabricated by either visual selection or by the lowest E* values determined from the photographs and a spectrophotometer. The same dental laboratory fabricated all 40 restorations. At the cementation appointment, clinical criteria were used to evaluate anatomy/contour, surface texture, and the amount of glaze as it relates to color perception before the restoration was cemented. RESULTS: The mean E* between the reference tooth before preparation and the crown before cementation in the visual assessment group was 10.49 (+/- 14.6), whereas the mean E* in the photocolorimetric group was 8.99 (+/- 5.7). Analysis of data showed that the observers and the colorimetric technique were perfect (E* = 0) 41% of the time and varied (E* = 0.1 or higher) 59% of the time. Data collected further showed no significant difference or correlation between shade selection methods and the evaluated clinical criteria. CONCLUSIONS: These results provide evidence that there is no significant difference in shade selection using the conventional visual assessment by two experienced clinicians or the photocolorimetric technique. CLINICAL SIGNIFICANCE: The use of photocolorimetric analysis in shade selection can serve as a reliable alternative to conventional visual shade selection. This method is useful for clinicians who have difficulty with shade selection. PMID- 12762477 TI - Talking with patients. Chairside CAD/CAM in dentistry. PMID- 12762478 TI - Radioprotective gene therapy. AB - Control of cancer by irradiation therapy alone or in conjunction with combination chemotherapy is often limited by organ specific toxicity. Ionizing irradiation toxicity is initiated by damage to normal tissue near the tumor target and within the transit volume of radiotherapy beams. Irradiation-induced cellular, tissue, and organ damage is mediated by acute effects, which can be dose limiting. A latent period follows recovery from the acute reaction, then chronic irradiation fibrosis (late effects) pose a second cause of organ failure. We have developed the technology for radioprotective gene therapy using the transgene for the antioxidant manganese superoxide dismutase, delivered to specific target organs (lung, esophagus, oral cavity, oropharynx, and bladder) using gene transfer vectors including plasmid/liposomes (PL) and adenovirus. Irradiation protection by MnSOD transgene overexpression at the cellular level has been demonstrated to be localized to the mitochondrial membrane. Using MnSOD transgene constructs lacking the mitochondrial localization leader sequence, and in other experiments attaching this localization signal to otherwise non-radioprotective cytoplasmic Cu/ZnSOD, mitochondrial localization has been demonstrated to be critical to protection. Organ specific injection of MnSOD-PL prior to irradiation demonstrates transgene expression for 48-72 hours, and an associated decrease in ionizing irradiation-induced expression of inflammatory cytokine mRNA and protein. Significant reduction of organ specific tissue injury has been demonstrated in several organ systems in rodent models. Application of MnSOD-PL gene therapy in the setting of fractionated chemo-radiotherapy is being tested in clinical trials for prevention of esophagitis during treatment of non-small cell carcinoma of the lung, and in prevention of mucositis during combination therapy of carcinomas of the head and neck. Encouraging results in pre-clinical models suggest that radioprotective gene therapy may facilitate dose escalation protocols to allow increases in the therapeutic ratio of cancer radiotherapy. PMID- 12762479 TI - Therapeutic angiogenesis in ischemic heart disease: gene or recombinant vascular growth factor protein therapy? AB - In the last decennium the challenge to research has been to find methods of inducing new vascular growth in ischemic myocardium due to atherosclerotic coronary artery disease, which could not be treated with balloon angioplasty or coronary artery by-pass grafting. Therapeutic angiogenesis with recombinant vascular endothelial growth factor proteins or gene encoding for the proteins is a new potential treatment for cardiovascular disease. The greatest interest and research has been concentrated on basic Fibroblast Growth Factor (FGF1 and FGF2) and Vascular Endothelial Growth Factor A (VEGF-A165 and VEGF-A121). Several small clinical phase I-II safety and efficacy trials with recombinant vascular endothelial growth factor proteins or gene encoding for the proteins have demonstrated that these treatment regimes seem to be safe and the results have been encouraging. However, two large doubleblind randomized placebo-controlled studies with intracoronary infusions of the recombinant proteins FGF2 and VEGF A165 could not detect any clinical effect. Large scaled phase II studies with gene therapy are in progress. Therapeutic angiogenesis is still a promising new treatment in patients with coronary artery disease. However, more research including large scaled clinical trials is needed before deciding whether the vascular endothelial growth factor therapy either as a gene or a recombinant slow release protein formulation therapy can be offered to patients with severe coronary artery disease, which cannot be treated with conventional revascularization. PMID- 12762480 TI - Tumor-specific gene delivery using genetically engineered bacteria. AB - The loco-regional control of cancer remains a major contributor to the treatment outcome for many cancer patients prescribed conventional radiotherapy or chemotherapy. Failure of treatment coupled with the realisation that cancer is essentially a genetic disease has led to development of many clinical protocols based on gene therapy. In this review, we will describe an alternative gene delivery system based on the use of non-pathogenic bacteria. Tumor regressions have been reported long ago in patients with bacterially infected tumors, suggesting that bacteria could target tumors and have local anti-tumor effects. The basis of this phenomenon is attributable to the unique properties of the tumor micro-environment. The presence of hypoxic and/or necrotic areas provides a haven for a number of anaerobic bacteria and over the past 60 years, several strains of anaerobic bacteria have been shown to localise within and cause cell lysis of experimental animal tumors. One of the most important strains in that context is Clostridium. Other bacteria have also been implicated in experimental anti-cancer settings. Of these, attenuated Salmonella strains capable of both selective amplification within tumors and expression of effector genes encoding therapeutic proteins are probably the most promising. We will discuss the potential advantages and the pitfalls of this alternative delivery approach. We will emphasize the importance of hypoxia in solid tumors and discuss the potential of radiation-inducible promoters and combined treatment modalities, involving vascular targeting and radiotherapy. We believe that this approach will act in a complementary way to current radiotherapy and chemotherapy treatments of solid tumors. PMID- 12762481 TI - Gene therapy of chronic pain. AB - Chronic pain is frequently associated with profound alterations of neuronal systems involved in pain processing and should be considered as an actual disease state of the nervous system. It should not only be relieved, but must really be treated in suffering patients. However, some forms of chronic pain, in particular those of neuropathic origin, are most often not satisfactorily managed with currently available pharmacological agents, some of which, in addition, may be poorly tolerated by some patients. In this context, gene-based approaches may contribute to the search for a better management of chronic pain. The question then arises regarding the most appropriate level for such an intervention using gene-transfer techniques. The first experimental protocols attempted the transfer of opioid precursor genes and their overexpression mainly at the spinal level. They demonstrated the feasibility and the real interest of these approaches by showing that local overproduction of opioid peptides induced antinociceptive effects in animal models of persistent pain, of inflammatory-, neuropathic- and even cancerous origin. Although really tempting data were obtained using gene based techniques in experimental inflammatory diseases, the possible clinical interest of these approaches in chronic pain has still to be established. Nevertheless, targeting some proinflammatory cytokines, involved not only in inflammation but also in the induction and probably the perpetuation of pain, raises the possibility to block the "development" of chronic pain rather that to "simply" relieve established ongoing pain. Future gene-based protocols will certainly target some of the recently identified molecules involved in pain transduction mechanisms, sensory nerve sensitization or pain perpetuation, and evaluate their potential interest to ideally abolish or, at least, reduce chronic pain. PMID- 12762482 TI - The pyruvate dehydrogenase complex as a target for gene therapy. AB - Here we review the rationale for considering the pyruvate dehydrogenase multienzyme complex (PDC) as a target for gene therapy for defects in mitochondrial energetics. PDC is entirely nuclear encoded and is situated in the mitochondrial inner membrane. The complex catalyzes the rate-determining step in aerobic carbohydrate metabolism and plays a critical role in the efficient conversion of substrate fuel into energy by cells. PDC activity is regulated in large part by reversible phosphorylation (inactivation) of its E1alpha subunit. Congenital defects in PDC are usually due to mutations in E1alpha and are typified by lactic acidosis, neurodegeneration and early death. Acquired deficiency in PDC has been implicated in the etiopathology of several other metabolic or neurodegenerative disorders. Recently, a vector using recombinant adeno-associated virus (rAAV) that contained a fusion protein of full-length E1alpha and the reporter gene green fluorescent protein was used to deliver wild type E1alpha into mitochondria after injection of the construct in vivo into the central nervous system of rats and in vitro into human cells. Transduction of cultured fibroblasts from a male patient with E1alpha deficiency led to partial restoration of PDC activity, as determined by decarboxylation of 14C-pyruvate. These data indicate that at least partial correction of PDC defects may be feasible by gene transfer. Furthermore, the combination of AAV-mediated delivery of E1alpha with pharmacologic activation (dephosphorylation) of the wild type enzyme subunit may provide an optimal therapeutic strategy for patients with acquired or congenital deficiencies in mitochondrial energy metabolism. PMID- 12762483 TI - Neuronal generation from somatic stem cells: current knowledge and perspectives on the treatment of acquired and degenerative central nervous system disorders. AB - Stem cell transplantation through cell replacement or as vector for gene delivery is a potential strategy for the treatment of neurodegenerative diseases. Several studies have reported the transdifferentiation of different somatic stem cells into neurons in vitro or after transplantation into animal models. This observation has pointed out the perspective of using an ethical and accessible cell source to "replace" damaged neurons or provide support to brain tissue. However, recent findings such as the cell fusion phenomenon have raised some doubts about the real existence of somatic stem cell plasticity. In this review, we will discuss current evidence and controversial issues about the neuroneogenesis from various sources of somatic cells focusing on the techniques of isolation, expansion in vitro as well as the inductive factors that lead to transdifferentiation in order to identify the factors peculiar to this process. The morphological, immunochemical, and physiological criteria to correctly judge whether the neuronal transdifferentation occurred are critically presented. We will also discuss the transplantation experiments that were done in view of a possible clinical therapeutic application. Animal models of stroke, spinal cord and brain trauma have improved with Mesenchymal Stem Cells or Bone Marrow transplantation. This improvement does not seem to depend on the replacement of the lost neurons but may be due to increased expression levels of neurotrophic factors, thus suggesting a beneficial effect of somatic cells regardless of transdifferentiation. Critical understanding of available data on the mechanisms governing the cell fate reprogramming is a necessary achievement toward an effective cell therapy. PMID- 12762484 TI - Sodium chloride modified silica nanoparticles as a non-viral vector with a high efficiency of DNA transfer into cells. AB - Development of reliable vectors is a major challenge in gene therapy. Previous gene transfer methods using non-viral vectors, such as liposomes or nanoparticles, have resulted in relatively low levels (35 to approximately 50%) of gene expression. We have developed a silicon nanoparticle (SNAP) system, a novel non-viral vector, for DNA transfer into cells. SNAP was synthesized chemically and modified with sodium chloride or sodium iodide. Electronmicroscopy of SNAP and fluorescence microscopy of fluorescence-labeled SNAP revealed that they were generated uniformly, had diameters of 10-100 nm, and showed a better efficiency (about 70%) of DNA transfection into cells as well as protection of DNA against degradation. The microscopy also demonstrated the adhesion of SNAP with HT1080 cell surface and entry of SNAP into the cells without cytotoxicity. Intravenous and/or intra-abdominal administration of the SNAP to mice revealed the accumulation of SNAP in the cells of the brain, liver, spleen, lung, kidney, intestine, prostate and the testis without any pathological cell changes or mortality, suggesting that they passed through the blood-brain, blood-prostate, and blood-testis barriers. These findings indicate that the SNAP generated has good biological characteristics as a potential promising vector for gene transfer, gene therapy and drug delivery. PMID- 12762485 TI - From the supply of POMs to proposed changes in the BVA's structure. PMID- 12762486 TI - Case-control study to investigate risk factors for horse falls in hurdle racing in England and Wales. AB - Between March 1, 2000 and August 31, 2001, a case-control study was conducted on 12 racecourses in England and Wales to identify and quantify the risk factors associated with horse falls in hurdle races. The cases and controls were defined so that variables relating to the horse, the jockey, the race and racecourse, and the jump could be considered. The cases were defined as a jumping effort at a hurdle flight that resulted in a fall, and the controls were defined as a successful jump over a hurdle at any of the 12 racecourses within 14 days before or after the case fall. Conditional logistic regression was used to examine the univariable and multivariable relationships between the predictor variables and the risk of falling. The risk of falling was significantly associated with the position of the jump in the race, and with the distance and speed of the race. A horse's previous racing experience and history were also significantly associated with the risk of falling and horses participating in their first hurdle race were at almost five times greater risk of falling than horses that had hurdled before. PMID- 12762487 TI - Evaluation of a dot ELISA kit for measuring immunoglobulin M antibodies to canine parvovirus and distemper virus. AB - A dot ELISA for the detection of immunoglobulin M (IgM) antibodies to canine distemper virus (CDC) and canine parvovirus (CPV) was assessed. The titres of IgM antibodies to CDV and CPV in 100 dogs were measured by the Immunocomb ELISA kit and compared with the results derived from the immunofluorescence assay (IFA). There was a strong correlation between the results of the dot ELISA technique and the IFA (P < 0.001). The dot ELISA kit was also used to assess the changes in the levels of immunoglobulin G (IgG) and IgM antibodies to CPV and CDV in 10 puppies vaccinated with a polyvalent vaccine. High levels of IgM antibodies to CPV were first detected seven days after they were vaccinated, and after nine days all the pups had high titres of IgG antibodies to CPV. High levels of IgM antibodies to CDV were detected after nine days and the highest average titres were recorded after 12 days. IgG antibodies to CDV were present from nine days after vaccination. PMID- 12762488 TI - Effects of anaesthesia and manual restraint on the plasma concentrations of pituitary and adrenocortical hormones in ferrets. AB - Two experiments were carried out to investigate the effect of sampling techniques on the plasma concentrations of pituitary and adrenocortical hormones in ferrets (Mustela putorius furo). In the first experiment blood was collected on two occasions from 29 ferrets which were either manually restrained or anaesthetised with isoflurane. In the second experiment eight intact ferrets were fitted with jugular catheters and blood was collected on four occasions, just before and as soon as possible after they had been manually restrained or anaesthetised with medetomidine or isoflurane; blood was also collected 10 and 30 minutes after the induction of anaesthesia. Medetomidine anaesthesia had no effect on the plasma concentrations of pituitary and adrenocortical hormones. Isoflurane anaesthesia resulted in a significant increase in the plasma concentration of alpha melanocyte-stimulating hormone (alpha-MSH) directly after the induction of anaesthesia. Manual restraint resulted in a significant increase in the plasma concentrations of cortisol and adrenocorticotrophic hormone (ACTH) and a decrease in the plasma concentration of alpha-MSH. PMID- 12762490 TI - Necrotic-ulcerative dermatitis on the heels of heifers in a dairy herd infected with Corynebacterium pseudotuberculosis. PMID- 12762489 TI - Surveillance of FMD virus non-structural protein antibodies in pig populations involved in an eradication programme. PMID- 12762491 TI - Effects of sainfoin hay on gastrointestinal nematode infections in goats. PMID- 12762492 TI - Crushed tail head syndrome in cattle. PMID- 12762493 TI - Control of bovine tuberculosis. PMID- 12762494 TI - Control of bovine tuberculosis. PMID- 12762495 TI - Equine passports. PMID- 12762496 TI - Fees for RCVS members, retired... PMID- 12762497 TI - Fees for RCVS members retired ... and new. PMID- 12762498 TI - Experiences of rabies. PMID- 12762499 TI - An overview of scientific and regulatory issues for the immunogenicity of biological products. AB - Immunogenicity of biological products can occur pre-clinically and clinically when products elicit immune responses in animals or humans receiving the products. This is a concern for manufacturers, regulatory agencies and clinicians as immune responses can result in effects on product effectiveness and safety. The clinical sequelae of immunogenicity range from no effects to serious, life threatening syndromes. However, although many biological products are immunogenic to some extent, it is quite rare that immunogenicity leads to serious adverse events. Whilst there are methods to detect immunogenicity, they currently rely on detecting the humoral rather than the cellular response of the immune system. The design and validation of assays such as immuno-assays and bio-assays are critical for a meaningful assessment of immunogenicity. There are a growing number of computational and laboratory-based methods for the prediction of immunogenicity, as well as methods to reduce potential immunogenicity and these may lead to less immunogenic biological products in future. PMID- 12762500 TI - Immunogenicity of biological therapeutics: a hierarchy of concerns. AB - CBER considers immune responses to biological therapeutic agents in a hierarchy, structured by clinical effects. The greatest concern regards immediate hypersensitivity responses that cause anaphylactic or anaphylactoid responses. Such responses have been most commonly observed in treatment with bacterial products such as asparaginase, streptokinase, and diptheria toxin-conjugated molecules. Immediate hypersensitivity, as well as more delayed hypersensitivity responses (hours to days) may also be observed in enzyme replacement therapies, wherein a normal mammalian enzyme appears as a foreign protein to deficient patients. More insidious, but nonetheless devastating, antibodies to a recombinant hormone or cytokine have been shown to neutralize not only the product, but also the endogenous factor. When the endogenous factor mediates a unique biological function, a clinical syndrome develops. Such has been observed in immune responses to recombinant erythropoietin and thrombopoietin, with patients exhibiting pure red blood cell aplasia and immune mediated thrombocytopaenia respectively. Of considerable importance, but posing less threat, is generation of binding antibodies which may cause infusion reactions, alter pharmacokinetics and biodistribution, and potentially diminish product efficacy. PMID- 12762501 TI - Immunogenicity of biopharmaceuticals. The European perspective. AB - In 1993 a Concerted EU Action was started to coordinate research concerning the immunogenicity of biopharmaceuticals bringing together researchers from academia and industry. The main topic of this concerted action has been interferon alpha. The factors influencing its immunogenicity were studied. Blind panel testing of patients' sera showed large differences in reported titres as an important variable in different studies. The EMEA has requested the three marketing authorization holders of interferon beta to develop a common assay for antibodies to this product. The status of this project will be discussed. The occurrence of about 30 cases of pure red cell aplasia related to antibodies induced by erythropoietin treatment will also be reported. PMID- 12762502 TI - Speculations on the immunogenicity of self proteins. AB - As recombinant proteins join the physician's armamentarium to a larger and larger extent, the capacity of these "self" proteins to induce an immune response has become more widely appreciated. The most surprising observation has been that these proteins may induce an immune response even in an individual who is not deficient in the protein, but merely produces an insufficient amount for the desired biological effect. Furthermore, individuals with no apparent predisposition to auto-immunity mount antibody responses to these proteins. Fortunately, the antibody response often does not block the activity of the proteins and sometimes the response may even disappear over time. Nevertheless, the immunogenicity of human recombinant proteins is a significant limitation in their use. This review first reviews aspects of B cell tolerance and activation and then presents a model to try to explain the auto-immune response to therapeutic proteins. PMID- 12762503 TI - Consequences of immunogenicity to the therapeutic monoclonal antibodies ReoPro and Remicade. AB - The clinical consequences of immune antibodies generated to abciximab (ReoPro) and infliximab (Remicade) are described. Abciximab, a chimaeric Fab fragment that binds to the beta3 integrin of the GPIIb/IIIa and alphavbeta3 receptors on human platelets, is approved in the US and Europe for use in percutaneous coronary intervention (PCI) to prevent cardiac ischaemic complications. The effects of induced antibodies upon the safety and efficacy of repeat administration of abciximab have been evaluated in the ReoPro Re-administration Registry Study, in which 5.7% of patients were HACA positive before re-treatment. An interim evaluation of 1000 patients has indicated that re-administration of abciximab can be accomplished in the setting of PCI with an acceptable safety and efficacy profile. Infliximab is a chimaeric IgG1 antibody specific for human TNFalpha, and is approved in the US and Europe for the acute treatment of the signs and symptoms of Crohn's disease and for the chronic treatment of rheumatoid arthritis (RA). The incidence of antibodies to infliximab is reported to be approximately 10%; however, an inverse dose-immunogenicity relationship was observed, indicating that higher doses of infliximab (> or = 3 to 10 mg/kg) could reduce the incidence of immune antibodies. The induction of immune antibodies could also be reduced by concomitant administration of low-dose methotrexate and other immunosuppressant agents. Although antibodies to infliximab appeared to be associated with lower serum infliximab concentrations and a slightly higher incidence of infusion reactions, these immune antibodies were generally not associated with a reduction in clinical efficacy. In addition, the antibodies induced to infliximab are specific for infliximab, and do not cross-react with other currently available therapeutic antibodies. PMID- 12762504 TI - Immunogenicity and immune tolerance coagulation Factors VIII and IX. AB - Some of the major issues related to the development and control of antibodies that occur during treatment of haemophilia with replacement factors (Factor VIII and Factor IX) are reviewed. Information on analytical issues, immunogenicity, and immune tolerance may be applicable to the study of other therapeutic proteins. Conversely, new information obtained from evaluation of other therapeutic protein products may address issues that remain unresolved for Factor VIII and FIX replacement therapy. PMID- 12762505 TI - Immunogenicity of GM-CSF products in cancer patients following immunostimulatory therapy with GM-CSF. AB - There is a high risk of developing neutralising and non-neutralising antibodies when GM-CSF is used as an immunomodulatory agent in non-immunocompromised patients. The presence of neutralising antibodies may seriously hamper the clinical response of the patients. This must be taken into account when designing protocols if the biological activity of the exogenously administered GM-CSF is not to be impaired and the endogenous production of GM-CSF is not to be inactivated. Assessment of production of neutralising antibodies during cytokine therapy is important for predicting the clinical response to progressive therapy. Use of validated assays is imperative for evaluation of antibodies generated following therapy with a particular protein. PMID- 12762506 TI - Modelling the immunogenicity of therapeutic proteins using T cell epitope mapping. AB - A new approach to designing therapeutic proteins is emerging, due to an improved understanding of T cell modulation of the immune response and new methods for modelling T cell epitopes using bioinformatics. In silico T cell epitope-mapping using the bioinformatics, when combined with other ex silico means of evaluating MHC-peptide and T cell interaction such as tetramers and HLA transgenic mice, enables the evaluation of dysfunctional immune responses to therapeutic proteins. This approach may even permit researchers to develop means of modulating anticipated adverse effects. The pocket profile method for developing T cell epitope prediction tools is reviewed here, and a comparison between the pocket profile method and the extended anchor method for epitopes restricted by the class II allele HLA DR B*0101 is described. PMID- 12762507 TI - The immunogenicity of biopharmaceuticals. Lessons learned and consequences for protein drug development. AB - Most biopharmaceuticals, including those proteins that are more or less identical to native human proteins, induce antibodies in a significant fraction of patients. The main factors contributing to immunogenicity are impurities and the presence of aggregates. Sequence divergence from the native proteins only plays a minor role except in proteins from microbial, plant or distant vertebrate origin. In the majority of cases the antibodies have no biological or clinical effects. The most common clinical effect is the loss of efficacy of the biopharmaceutical. Serious complications of immunogenicity are rare. The best method to prevent immunogenicity is optimizing production, purification and formulation of the biopharmaceutical protein to generate soluble, non-aggregated, native protein free of contaminating adjuvants. The best way to predict immunogenicity in humans is evaluation in immune tolerant transgenic mice. PMID- 12762508 TI - Use of cell-based therapies for modification of host immune responses. AB - The success of adoptive cellular therapy depends on the ability to select optimally or produce cells genetically with the desired antigenic specificity, and then induce cellular proliferation while preserving the effector function, engraftment, and homing abilities of the lymphocytes. Unfortunately, many previous clinical trials were carried out with adoptively transferred cells that were propagated in what are now understood to be sub-optimal conditions that impair the essential functions of the adoptively transferred cells. This article reviews some of the lessons and developments emerging from the past 20 years of adoptive immunotherapy trials and basic immunology regarding immunogenicity, T cell homeostasis, and the maintenance of tolerance and repertoire. PMID- 12762509 TI - Vailidation of immunoassays for anti-drug antibodies. AB - Several methods for the detection of an antibody response to a protein therapeutic are described along with their limitations. Emphasis is placed on immunoassays with specific attention to the ELISA format. Potential sources of interference and issues to consider in the design of an anti-drug antibody assay are reviewed. Establishment of criteria for validation of an anti-drug assay are discussed including inter-assay and intra-assay precision for the positive and negative controls, limit of detection, incidence of false positives, reactivity in unexposed populations, linearity of dilution, and interference issues. Finally, a case study involving the development of an anti-host cell protein contaminant assay involving responses to Chinese hamster ovary (CHO) cell contaminants is reviewed. Detection of an immune response to a biotherapeutic is critical to an understanding of its' safety and efficacy. Outlined in this paper are standard parameters for validation of an immunoassay as well as practical considerations for the development and validation of an anti-protein antibody assay. PMID- 12762510 TI - Bioassays for the measurement of neutralizing antibodies in serum (SNFs). AB - Bioassays to measure serum neutralizing factor (SNF) activity against therapeutic biological products are an important tool for assessment of the immune status of patients on a dosing regimen. Because of the large diversity of bioanalytical methodology currently available to measure neutralization of a biological response, approaches to assay development and validation can create a considerable challenge. Assay design can take on many forms depending on the biological response to be monitored and the severity of complicating serum effects. In addition, the manner in which assay results are reported should have clinical relevance, (e.g. related back to the amount of analyte neutralized per volume of serum, etc.). Although ICH/FDA assay validation guidelines exist, they were not created with biotechnology-based bioassays in mind. Therefore, a thorough assessment of the analytical method and proper interpretation of current guidelines is critical for a meaningful validation effort. Several assay development strategies and approaches to assay validation will be discussed. PMID- 12762511 TI - New technologies for the detection of antibodies to therapeutic proteins. AB - The determination of antibody formation in response to a therapeutic product requires that a variety of data be evaluated. In addition to examining immuno assay and bioassay results, it is important to determine any clinical relevance. By also examining pharmacokinetic data as well as other clinical results, an evaluation can be made as to whether any clinically relevant antibodies were generated. To help in this evaluation, it is important to fully characterize the antibodies that are generated. Both the BIAcore and IGEN platforms are powerful tools for determining if antibodies have been produced in a subject The BIAcore is also able to characterize these antibodies as to isotype, relative concentration, and relative affinity. The bioassay result can identify serum samples that contain an agent capable of inhibiting a biological effect of a drug. When combined with the results of an immuno-assay, it is possible to determine if the antibodies are capable of neutralizing the drug. PMID- 12762512 TI - Animal models as indicators of immunogenicity of therapeutic proteins in humans. AB - Animal models have not been able to predict the immunogenicity of therapeutic proteins in humans reliably. The main issue is that administration of a human protein in an animal species is likely to be immunogenic. In non-human primate studies, we have seen a variety of responses; from little to no antibody response, to a strong neutralizing response, or even a cross-reactive antibody response. These have generally not correlated well with the immune response seen in humans. The route of administration, duration and schedule of dosing, the cumulative dosage of the protein, the pharmacological (i.e., immune altering) properties of the protein, as well as the purity of the clinical material can influence the immunogenicity. The animal studies should mimic these factors to the best extent possible for the animal model to be at all relevant to humans. Models do exist which provide valuable information to compare the immunogenicity of various compounds or routes of administration. Presumably, this 'relative' immunogenicity would be similar in humans. Additional characterization of transgenic mice, or use of homologous proteins, may help to establish better models to predict immunogenicity. PMID- 12762513 TI - Improving the quality of BIACORE-based affinity measurements. AB - Although the BIACORE technology has become the standard method for measuring the affinity of antigen-antibody interactions, many users of the technology do not apply the most advanced experimental design and data processing methods that are now available. In addition, many published reports fail to provide the experimental details that are necessary to assess the reliability of the affinity data that are presented. This review describes the experimental conditions that should be used to ensure that reliable biosensor data are collected. There is little justification for the belief that ELISA methods provide in principle more reliable affinity parameters than biosensor technology. PMID- 12762514 TI - Standardization of antibody preparations for use in immunogenicity studies: a case study using the World Health Organization International Collaborative Study for Islet Cell Antibodies. AB - The immunogenicity of biological therapeutic products is currently a high profile regulatory and biotechnology industry issue. The immune responses raised against biotechnology products range from the benign, to affecting product efficacy, to those that have serious deleterious clinical impact. The most widely used marker of immunogenicity is the detection and measurement of antibody responses induced in vivo to a product. This relies on assays that are sensitive and robust. In order to assess the parameters of an assay during its design, development and validation, it is extremely useful to have a reference standard to compare assay results. However, immune responses lead to polyclonal antibody preparations that can vary by affinity and avidity. This makes it extremely difficult to select a preparation that will behave similarly in different test systems and against different antibody samples. The case example of the WHO standardization of islet cell antibodies illustrates the difficulties in the process and the mechanisms required to produce a suitable antibody standard. PMID- 12762515 TI - Establishing personal identification based on specific patterns of missing, filled, and unrestored teeth. AB - The primary goal of this research is to examine the overall utility of nonradiographic dental records for the establishment of individual identifications. It was found that even without radiographic lines of comparison, charts and notes that accurately detail a missing individual's antemortem dental condition can be essential for establishing an identification. Based on an analysis of two large datasets, individual dental patterns were determined to be generally unique, or at least very uncommon. Through this type of empirical comparison, it is possible to establish a strong, quantifiable association with a missing individual. The results of this research indicate that a definitive number of points of concordance do not need to be established in dental identification cases. Each case must be assessed individually. The critical factor is to remove subjective judgment calls from dental comparisons. This research has proposed a new method of empirical comparison that allows forensic odontologists to derive objective frequency information regarding the occurrence of specific dental patterns in the general population. The method is similar to that used for mtDNA casework, and a computer program (OdontoSearch) has been developed to make the technique accessible. It was found that even a small number of common dental characteristics may produce a very rare dental pattern, a point that may be counterintuitive to many forensic odontologists. PMID- 12762516 TI - The diversity of adult dental patterns in the United States and the implications for personal identification. AB - Few would argue about the individuality of dental radiographs for forensic identification, but when an antemortem/postmortem comparison is based strictly on dental treatment notes and/or charts it becomes less certain. In the past, attempts to validate the high diversity of dental patterns created by combinations of missing, filled, and unrestored teeth have been based on unfounded statistical assumptions. The goal of this research is to present a statistically valid method of assessing dental pattern diversity for the identification of missing individuals. Empirical observation of large reference datasets was found to be the best technique for assessing dental diversity. This technique is nearly identical to the procedure used for mitochondrial DNA casework. For the research presented in this paper, two large datasets were used, one composed of U.S. military personnel and one composed of U.S. civilians. Dental patterns were found to be very diverse on a scale that is comparable to mtDNA. In addition, it was found that the diversity values remain very consistent regardless of the level of detail present in the treatment records. Overall, combinations of missing, filled, and unrestored teeth were found to be very individualistic and an excellent source for forensic identification. PMID- 12762517 TI - Estimation of age in adolescents --the basilar synchondrosis. AB - The state of fusion of the basilar synchondrosis as a biological age indicator was assessed in a sample of 91 cadavers of both sexes whose ages ranged between 8 and 26 years. The correlation between the degree of closure and chronological age was investigated. Although the female population sample was very small (n = 21), the data indicate a tendency of differences in age between the "open" and "closed" groups. In the male population (n = 70), no significant differences were detected between the "open" and "closed" categories; in fact, the mean age of the two groups was the same (p = 0.9). These findings indicate that the stage of fusion of the basilar synchondrosis is not a good indicator of age in male cadavers, while in females the feature could be useful when estimating age of unknown human remains, although further investigation on a larger sample is advocated. PMID- 12762518 TI - Discriminant function sexing of fragmentary and complete femora: standards for contemporary Croatia. AB - Determining sex is one of the first and most important steps in identifying decomposed corpses or skeletal remains. Previous studies have demonstrated that populations differ from each other in size and proportion and that these differences can affect metric assessment of sex. This paper establishes standards for determining sex from fragmentary and complete femurs in a modern Croatian population. The sample is composed of 195 femora (104 male and 91 female) from positively identified victims of the 1991 War in Croatia. Six discriminant functions were generated. one using seven variables, three using two variables, and two employing one variable. Results show that complete femora can be sexed with 94.4% accuracy. The same overall accuracy, with slight differences in male/female accuracy, was achieved using a combination of two variables defining the epiphyses, and with the variable maximum diameter of the femoral head. PMID- 12762519 TI - The external occipital protuberance: can it be used as a criterion in the determination of sex? AB - Sex determination of a corpse can be problematic in cases where the body is damaged. Useful criteria would assist in the identification of sex in such cases. The goal of this study is to determine the usefulness of the external occipital protuberance (EOP) in the determination of sex, especially in lateral cranium radiographs. The types and configurations of the EOP were investigated on normal lateral cranium radiographs of 1000 subjects (500 males and 500 females) and 694 dry-skull remains (371 males and 323 females) from a 16th Century Anatolian population for the purpose of sex determination. In the radiographic examination, the incidence of less prominent (Type 1) EOP is found to be 85.4% in females whereas 17.8% in males. The spine type (Type 3) EOP is found to be 63.4% in males and to be 4.2% in females. On the other hand, studies of dry-skull remains revealed the incidence of Type 1 EOP to be 67.5% in females and Type 3 EOP to be 55.2% in males. The crest type (Type 2) EOP is approximately equal in both sexes and is found to be less valuable for sex determination in both groups. PMID- 12762520 TI - Reexamination of a measurement for sexual determination using the supero-inferior femoral neck diameter in a modern European population. AB - The present study reexamines the accuracy of the supero-inferior femoral neck diameter for the determination of sex using a modern sample of French individuals. In 1998, Seidemann et al. used this univariate method for sex determination with the Hamann-Todd collection. Stojanowski and Seidemann in 1999 tested previous results on a modern sample taken from the University of New Mexico and concluded that the Caucasian male samples exhibited no significant differences between individuals born before and after 1900, but the Caucasian female subgroup did exhibit differences with an increase of the SID in the modern sample. The current study compares the previous results of the supero-inferior femoral neck diameter with a modern sample of elderly French individuals born after 1910. Both sides of the femur were measured. No statistical difference was found between the right and left side (p = 0.31). The results showed a significant difference between the pre-1900 and the modern sample, with an increase in femoral neck diameter in modern populations. The comparison of the SID values between the two modern samples (Mexico and Nice) showed no significant differences in the femoral neck diameter in the two male subgroups (p = 0.05), but the measurements of the SID in the female subgroup did exhibit significant differences with an increase of the neck femoral diameter (p < 0.01) in the modern French population. These results demonstrate an increase in the neck femoral morphology in the elderly European French females samples. PMID- 12762522 TI - Mummified trophy heads from Peru: diagnostic features and medicolegal significance. AB - Several forms of mummified human trophy heads were produced by prehistoric and historic native groups in South America. This paper describes the diagnostic features of trophy heads produced by the Nasca culture of ancient Peru. A growing interest in these mummified heads among collectors of Pre-Columbian art and antiquities has led to their illegal exportation from Peru, in violation of national and international antiquities laws. Requests from the Peruvian government to protect its cultural patrimony led the United States in 1997 to declare these heads as items subject to U.S. import restriction, along with six other categories of human remains. Despite such restrictions, Nasca trophy heads continue to reach private collectors outside of Peru and thus may be encountered by local, state, or federal law enforcement officials unfamiliar with their characteristic features and origin. The objective of this paper is to describe the features that allow Nasca trophy heads to be identified and distinguished from other archaeological and forensic specimens that may be submitted to a forensic anthropologist for identification. PMID- 12762521 TI - Twenty-seven years of forensic anthropology casework in New Mexico. AB - A review of anthropological consult cases for the New Mexico Office of the Medical Investigator was conducted for the years 1974 through 2000. A total of 596 cases are summarized and information is presented on the sex and age of the individuals, season of recovery, depositional environment, body covering, time since death, perimortem trauma, postmortem animal activity, and skeletal element recovery. Results reveal a higher percentage of male victims (76%). No variation is seen in the seasonal distribution of cases. In cases with known time since death, 35% were recovered within one week while 30% had a postmortem interval exceeding one year. Depositional environments include surface (45%), burial (13%), and airplane crashes (9.5%). In 42% of the cases, no evidence of perimortem trauma was observed. Postmortem animal activity was noted in 46% of cases. Data presented in this study may prove useful in supporting expert witness testimony and generating future research models. PMID- 12762523 TI - Characterization of pyrotechnic reaction residue particles by SEM/EDS. AB - Today the method commonly used for detecting gunshot residue is through the combined use of scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS). In recent years, this same methodology began to find use in detecting and characterizing pyrotechnic reaction residue (PRR) particles whether produced by explosion or burning. This is accomplished by collecting particulate samples from a surface in the immediate area of the pyrotechnic reaction. Suspect PRR particles are identified by their morphology (typically 1 to 20 microm spheroidal particles) using an SEM; then they are analyzed for the elements they contain using X-ray EDS. This can help to identify the general type of pyrotechnic composition involved. PMID- 12762524 TI - Glass-containing gunshot residue particles: a new type of highly characteristic particle? AB - In 0.22 caliber rimfire ammunition, the primer often contains lead or lead and barium compounds. As residues from these primers do not contain lead, barium, and antimony, they cannot be uniquely classified as gunshot residue (GSR) under ASTM designation E 1588-95. In many types of 0.22 caliber rimfire ammunition, the cartridge contains a primer sensitized with glass. In this paper we describe a previously unreported type of GSR particle consisting of glass fused with other primer components. As there appear to be few potential environmental or occupational sources of particles composed of lead and barium compounds fused to glass, particularly borosilicate glass, these particles may have high evidential value. Scanning electron microscopy with energy dispersive X-ray detection (SEM EDX) and time-of-flight secondary ion mass spectrometry (TOF-SIMS) were evaluated for the characterization of glass-containing GSR particles. The occurrence of glass-containing GSR particles was established in the residue from various brands of 0.22 caliber ammunition, and several sub-types were identified. PMID- 12762525 TI - Characterization of surface organic components of human hair by on-line supercritical fluid extraction-gas chromatography/mass spectrometry: a feasibility study and comparison with human identification using mitochondrial DNA sequences. AB - This paper discusses results of a supercritical fluid extraction-gas chromatography/mass spectrometry (SFE-GC/MS) study of small samples ( 100 microg to 1 mg) of human scalp hair. The method offers a number of benefits including greater sensitivity than liquid extraction methods because the entire extractable mass is transferred to the analytical system, compared with only a few percent from a conventional liquid extraction/injection. The project's goals were to determine if SFE-GC/MS analyses of the surface-extractable components of an individual's hair yield consistent chemical profiles and to investigate if the profiles are sufficiently different to distinguish them from those of other individuals. In addition, the mtDNA sequences from ten of the same individuals used in the SFE-GC/MS study from four family units were determined, and, while the families were distinguishable, the maternal relations yielded identical sequences. In tandem, SFE-GC/MS and mtDNA techniques may provide valuable complementary data from forensic hair samples. PMID- 12762526 TI - ESDA processing and latent fingerprint development: the humidity effect. AB - The influence of humidification in the ESDA process on subsequent development of fingerprints on paper items was studied. It was found that, while the DFO process is nearly insensitive to previous humidification, fingerprint development with ninhydrin or with indanedione can be significantly affected by previous humidification of the paper. PMID- 12762527 TI - Evaluation of a multicapillary electrophoresis instrument for mitochondrial DNA typing. AB - Laser-induced detection of fluorescent labeled PCR products and multi-wavelength detection (i.e., multicolor analysis) enables rapid generation of mtDNA sequencing profiles. Traditionally, polyacrylamide slab gels have been used as the electrophoretic medium for mtDNA sequencing in forensic analyses. Replacement of slab gel electrophoresis with capillary electrophoresis (CE) can facilitate automation of the analytical process. Automation and high throughput can be further enhanced by using multicapillary electrophoretic systems. The use of the ABI Prism 3100 Genetic Analyzer (ABI 3100, Applied Biosystems, Foster City, CA) as well as the ABI Prism 310 Genetic Analyzer (ABI 310, Applied Biosystems, Foster City, CA) were evaluated for mtDNA sequencing capabilities and compared with sequencing results obtained on the platform currently in use in the FBI Laboratory (the ABI Prism 377 DNA Sequencer, ABI 377, Applied Biosystems, Foster City, CA). Various studies were performed to assess the utility of the ABI 3100, as well as the ABI 310 for mtDNA sequencing. The tests included: comparisons of results obtained among the ABI 3100, the ABI 310 and the ABI 377 instruments; comparisons of results obtained within and between capillary arrays; evaluation of capillary length; evaluation of sample injection time; evaluation of the resolution of mixtures/heteroplasmic samples; and evaluation of the sensitivity of detection of a minor component with reduced template on the ABI 3100. In addition, other studies were performed to improve sample preparation; these included: comparison of template suppression reagent (TSR, Applied Biosystems, Foster City, CA) versus formamide; the use of Performa DTR Gel Filtration Cartridges (Edge BioSystems Inc., Gaithersburg, MD) versus Centri-Sep Spin Columns (Princeton Separations, Adelphia, NJ) for product purification after cycle sequencing; and sample stability after denaturation. The data support that valid and reliable results can be obtained using either capillary electrophoresis instrument, and the quality of sequencing results are comparable to or better than those obtained from the ABI 377 instrument. PMID- 12762528 TI - Further studies on spot tests and microcrystal tests for identification of cocaine. AB - The presence of cocaine in illicit drug samples is still being determined in some laboratories using spot tests and microcrystal tests. Seventeen chemical species were tested using three different spot tests (Wagner, Marquis, and cobalt thiocyanate followed by stannous chloride reactions) and two microcrystal tests (gold chloride and platinic chloride) to determine whether the results could be differentiated from the results of these tests on cocaine. The data obtained indicated that nine of the 17 compounds gave results similar to those from cocaine using the three spot tests, but that the results from microcrystal testing allowed for differentiation of all nine compounds from cocaine. PMID- 12762529 TI - Fingerprints as evidence for a genetic profile: morphological study on fingerprints and analysis of exogenous and individual factors affecting DNA typing. AB - Material recovered from 374 fingerprints left by eleven laboratory workers on three different substrates (glass, wood, metal) at a standard pressure time of 30 s, with and without preliminary handwashing, was submitted to morphological, quantitative, and type analysis. Morphological and agarose-gel electrophoresis analysis showed that a non-negligible amount of epidermal corneal cells presented apoptotic alterations. The quantity of DNA recovered from fingerprints ranged between 0.04 to 0.2 ng, and in a significant number of experiments no DNA was detected. Handwashing reduced the amount of DNA recovered from fingerprints. The "shedder status" of the donor was a very important factor, causing inter individual variations in the amount of DNA left by fingerprints. Spurious alleles from laboratory-based and secondary transfer contamination, stutters, and other artifacts described when analyzing low-copy-number DNA and capable of affecting correct profiles were observed. PMID- 12762530 TI - Background correction in forensic photography. I. Photography of blood under conditions of non-uniform illumination or variable substrate color--theoretical aspects and proof of concept. AB - The combination of photographs taken at two or three wavelengths at and bracketing an absorbance peak indicative of a particular compound can lead to an image with enhanced visualization of the compound. This procedure works best for compounds with absorbance bands that are narrow compared with "average" chromophores. If necessary, the photographs can be taken with different exposure times to ensure that sufficient light from the substrate is detected at all three wavelengths. The combination of images is readily performed if the images are obtained with a digital camera and are then processed using an image processing program. Best results are obtained if linear images at the peak maximum, at a slightly shorter wavelength, and at a slightly longer wavelength are used. However, acceptable results can also be obtained under many conditions if non linear photographs are used or if only two wavelengths (one of which is at the peak maximum) are combined. These latter conditions are more achievable by many "mid-range" digital cameras. Wavelength selection can either be by controlling the illumination (e.g., by using an alternate light source) or by use of narrow bandpass filters. The technique is illustrated using blood as the target analyte, using bands of light centered at 395, 415, and 435 nm. The extension of the method to detection of blood by fluorescence quenching is also described. PMID- 12762531 TI - Background correction in forensic photography. II. Photography of blood under conditions of non-uniform illumination or variable substrate color--practical aspects and limitations. AB - The combination of photographs taken at wavelengths at and bracketing the peak of a narrow absorbance band can lead to enhanced visualization of the substance causing the narrow absorbance band. This concept can be used to detect putative bloodstains by division of a linear photographic image taken at or near 415 nm with an image obtained by averaging linear photographs taken at or near 395 and 435 nm. Nonlinear images can also be background corrected by substituting subtraction for the division. This paper details experimental applications and limitations of this technique, including wavelength selection of the illuminant and at the camera. Characterization of a digital camera to be used in such a study is also detailed. Detection limits for blood using the three wavelength correction method under optimum conditions have been determined to be as low as 1 in 900 dilution, although on strongly patterned substrates blood diluted more than twenty-fold is difficult to detect. Use of only the 435 nm photograph to estimate the background in the 415 nm image lead to a twofold improvement in detection limit on unpatterned substrates compared with the three wavelength method with the particular camera and lighting system used, but it gave poorer background correction on patterned substrates. PMID- 12762532 TI - High mortality rates among inmates during the year following their discharge from a French prison. AB - While the poor health status of prisoners has been highlighted in Western countries, the surveillance of their mortality has been neglected. We studied the mortality of 1305 prisoners released during 1997 from a French prison. Vital status after release was obtained for 86.4% of them. Compared with the general population, ex-prisoners non-natural mortality rates were significantly increased both in the 15-34 and 35-54 age categories (3.5-fold and 10.6-fold respectively) and the risk of death due to overdose was 124 and 274 times higher in the same categories respectively. Prevention and care should be reinforced in the pre release period without waiting more epidemiological data. PMID- 12762533 TI - Cadaver dog and handler team capabilities in the recovery of buried human remains in the southeastern United States. AB - The detection of human remains that have been deliberately buried to escape detection is a problem for law enforcement. Sometimes the cadaver dog and handler teams are successful, while other times law enforcement and cadaver dog teams are frustrated in their search. Five field trials tested the ability of four cadaver dog and handler teams to detect buried human remains. Human and animal remains were buried in various forested areas during the summer months near Tuscaloosa, Alabama. The remains ranged in decomposition from fresh to skeletonized. Cadaver dogs detected with varying success: buried human remains at different stages of decomposition, buried human remains at different depths, and buried decomposed human and animal remains. The results from these trials showed that some cadaver dogs were able to locate skeletonized remains buried at a significant depth. Fresh and skeletonized remains were found equally by the cadaver dogs along with some caveats. Dog handlers affected the reliability of the cadaver dog results. Observations and videotape of the cadaver dogs during field trials showed that they were reliable in finding buried human remains. PMID- 12762534 TI - Quantification of forensic DNA from various regions of human teeth. AB - When the use of traditional forensic identification methods such as fingerprints or dental radiographs is difficult or impossible, identification by DNA analysis has proven valuable. In situations such as explosions or airplane crashes, identification is even more difficult because human remains are often fragmented and may be commingled. Teeth are a useful source of DNA and can often survive extreme environmental conditions. However, teeth may be fragmented into several identifiable regions. Therefore it is important to determine if DNA is present in forensically significant yields in all regions of the tooth. The main objectives of this study were to determine which region(s) of the tooth contains quantifiable DNA, if all regions contain similar yields of DNA and whether there is enough DNA in all regions to justify DNA extraction from a found tooth fragment. Results demonstrate that there is sufficient quantity of DNA in the crown body, root body, and root tip to support DNA extraction. Additionally, the root body is the region with the highest yield of DNA. This information will aid forensic DNA analysts in producing a useful DNA profile in a timely and cost effective manner. PMID- 12762535 TI - Diffuse vascular injury in fatal road traffic accident victims: its relationship to diffuse axonal injury. AB - The authors have reported a macro- and microscopic study of brain lesions in 120 victims of fatal road traffic accidents, independent of the survival time. Diffuse vascular injury (DVI) was found in 14 patients (11.7%). All patients with DVI died within 24 h after the accident. The 14 patients with DVI also showed severe (Grade 2 or 3) diffuse axonal injury (DAI). Since DVI is restricted to road traffic accidents and incompatible with life, the high frequency observed in our series could be explained by the fact that all 120 patients were victims of road traffic accidents, and 69.2% had died within 24 h after the accident. The association between DVI and severe DAI (Grades 2 and 3) suggests that both lesions depend on the same mechanism, with the degree of axonal and vascular damage being determined by the intensity of the head acceleration. Our results show a relationship between DVI and DAI that suggest there may be a spectrum or at least a continuum between these entities as distinct from DVI being a separate entity. PMID- 12762536 TI - Evidence of hexavalent chromium ingestion. AB - We describe the application of histochemical demonstration of chromium in a case of fatal ingestion of potassium dichromate in a suicide attempt. Using 2-(8 quinolylazo)-4,5-di-p-tolylimidazole (QTI), we could demonstrate chromium in the erythrocyte of the victim, in situ. This finding provides a means of proving the hexavalent chromium ingestion. PMID- 12762537 TI - Stability of cholesterol gall stones after 165 years of burial. AB - A woman who died in 1837 was exhumed for the purposes of moving the grave to another location. During the excavation, small white deposits of stone were uncovered in the right abdominal region, inferior to the rib cage and superior to the ilium blade. These stones were analyzed for cholesterol, bilirubin, and calcium following solubilization using methyl tert-butyl ether as a solvent. The results of these clinical chemistry analyses showed that these stones consisted primarily of cholesterol. Under these particular soil conditions encountered in this case, cholesterol gall stones are stable for at least 165 years. PMID- 12762538 TI - Investigation of Italy's deadliest building collapse: forensic aspects of a mass disaster. AB - We describe the investigation of the 1999 collapse of an apartment building in Foggia, Italy. Sixty-one victims were recovered in the rubble of the building, and five people were unaccounted for. All the bodies were well preserved except for two who had been burned. The majority of the victims were identified visually or by comparing body features, clothing, or personal effects with information collected from relatives or friends. Positive identifications of the two victims who were burned were obtained by dental comparison and DNA analysis. Approximately half of the victims (51.6%) sustained fatal injuires, while the remainder died from asphyxia. The injuries were characterized using the Abbreviated Injury Scale (AIS) and the New Injury Severity Score (NISS) systems. Injury severity associated with the location of victims inside the apartment may provide useful information for those involved in building design and/or search and rescue operations. Engineers determined that the collapse was the result of the use of inappropriate foundation material. PMID- 12762539 TI - Cutaneous current marks due to a stun gun injury. AB - Histological changes of the skin following electrical injury with a stun gun have rarely been described. We report the case of a 61-year-old man who died after having been tortured with a stun gun during a robbery. At autopsy two reddish, dot-like lesions where found on the chest and histological examination revealed electric current-related changes. Only a few reports concerning micromorphological cutaneous changes following stun gun injury have been reported; therefore further investigations concerning the frequency and type of histological findings due to stun gun injuries will be necessary in order to provide sufficient characteristic data for a conclusive interpretation. PMID- 12762540 TI - Exsanguination by subclavian-esophageal fistula associated with a left subclavian aneurysm. AB - Aortic aneurysms are a common autopsy finding, but aneurysms confined exclusively to the subclavian arteries are rare. When found, they are typically associated with trauma, surgery, or aberrant vessel distribution. Subclavian-esophageal fistula formation is also rare, with the vast majority being related to aberrant vessel distribution or esophageal foreign bodies. Dicle et al. first reported a subclavian-esophageal fistula associated with a non-aberrant subclavian artery aneurysm in 1999 (1). The following case would mark the second report of that phenomenon, and the first in the setting of a forensic autopsy. PMID- 12762542 TI - Photodegradation and laser desorption mass spectrometry for the characterization of dyes used in red pen inks. AB - Photodegradation and laser desorption mass spectrometry (LDMS) is a powerful combination of methods capable of characterizing dyes found in pen inks. Rhodamine dyes in pens that contain red ink were analyzed directly from paper (no extraction step is necessary). Inks exposed to incandescent light form photodegradation products (compounds with lower molecular weights than that of the intact dye) and in some instances, photoproducts (compounds with higher molecular weights than that of the intact dye). The degradation products and photoproducts can be detected with LDMS, and the results can be used for dye identification. Advantages include: (1) the instrumental analysis takes less than a minute; (2) sample preparation is minimal; (3) LDMS is a minimally destructive technique; (4) incandescent light sources are inexpensive, safe to use, and readily available; and (5) isomeric dyes can be distinguished. PMID- 12762541 TI - Measures of aggression and mood changes in male weightlifters with and without androgenic anabolic steroid use. AB - Supraphysiologic doses of testosterone are associated with increased aggression that is hypothesized to be a function of testosterone serum concentrations, mood, and personality. The study attempted to characterize this relationship among weightlifters who were users (n = 10) and nonusers (n = 18) of anabolic steroids. Participants were interviewed using the Modified Mania Rating Scale and Hamilton Rating Scale for Depression to assess mood, the Buss-Durkee Hostility Inventory (BDHI) and Point Subtraction Aggression Paradigm (PSAP) to assess aggression, and the Personality Disorder Questionnaire (PDQ-R) to assess personality. Blood samples were obtained for the determination of total, free, and weakly bound testosterone. Comparisons of continuous variables between testosterone users and non-users were performed with a parametric (unpaired t-test) or non-parametric (Mann-Whitney) test where appropriate. Correlations with testosterone were examined separately for testosterone users and non-users, using Spearman rank correlation. The subjective (BDHI) and objective (PSAP) assessments of aggression found that supranormal testosterone concentrations were associated with increased aggression. However, the PDQ-R results suggest that this finding was confounded by the personality disorder profile of the steroid users, because steroid users demonstrated Cluster B personality disorder traits for antisocial, borderline, and histrionic personality disorder. PMID- 12762543 TI - The effects of latent print processing on questioned documents produced by office machine systems utilizing inkjet technology and toner. AB - Counterfeiting of currency and identity documents, death threats, illegitimate business transactions, and terrorist-related activities are some examples of the types of crimes that often involve documents produced from printers and copiers. Although standard protocol typically requires a questioned document (QD) examination prior to latent print (LP) processing, occasionally, items of evidence may be submitted for a QD examination following the application of a series chemicals utilized in the development of latent fingerprints. In such cases, the forensic examiner must take into account any previous treatments prior to initiating an examination on documents produced with a printer or copier. This study was devised to examine the effects of a latent print development technique [ninhydrin, physical developer, and a bleach enhancer] on the physical and chemical examination of documents produced from copiers and printers. PMID- 12762544 TI - Codeine and morphine blood concentrations increase during blood loss. AB - During extensive blood loss, a plasma volume refill will take place by transfer of extravascular fluid into the circulation. Drugs present in this fluid may follow and cause a rise or a drop in blood drug concentration, depending on their levels and accessibility in the restoration fluid. This study explored the possible changes of codeine, and its metabolite morphine, in whole blood during a standardized exsanguination in the rat. Three doses containing 5 mg codeine were given orally. In eight rats, blood loss was accomplished by slowly withdrawing 0.8 mL blood at 10 min intervals during 70 min. In control rats, blood was withdrawn only at 0 and 70 min. At 70 min, the final/initial codeine and morphine concentration ratios were 0.70 +/- 0.38 and 0.88 +/- 0.47, respectively, in controls, but increased to 1.28 +/- 0.44 (p = 0.014) and 1.41 +/- 0.34 (p = 0.021), respectively, in exsanguinated rats. It is concluded that blood loss can affect blood drug concentrations. PMID- 12762545 TI - Methadone-related deaths in Hennepin County, Minnesota: 1992-2002. AB - Methadone maintenance therapy (MMT) is the only currently established medical therapy for heroin addiction. However, MMT still remains controversial. In Hennepin County, Minnesota, methadone is one of the top ten drugs reported in medical examiner investigated deaths and one of the most commonly diverted pharmaceuticals. This report reviews the role of methadone in medical examiner deaths over a 10-year period, 1992-2002. We compare cause and manner of death (accidental, natural, suicide) and methadone blood concentrations for decedents who were members of MMT programs with illicit users and those prescribed methadone for chronic pain. Findings reveal that 65% of decedents with measurable blood methadone concentrations were not participating in MMT programs. A total of 96 cases were identified, with the majority white (90.5%) and male (76.8%). MMTP program members were the minority (34.7%) of the methadone positive deaths and 39% were illicit users. Fifteen percent were chronic pain patients with almost half of this group dying from overdose. Methadone concentrations of drug caused/related deaths (0.18-3.99 mg/L) overlapped with those of deaths not attributable to methadone (0.18-3.03 mg/L) with no definable lethal level. Interpretation of methadone blood concentrations must be done in the context of the clinical history for determining cause of death, and may be confounded by postmortem redistribution. PMID- 12762546 TI - Improper sealing caused by the Styrofoam integrity seals in leakproof plastic bottles lead to significant loss of ethanol in frozen evidentiary urine samples. AB - Evidentiary urine samples (n = 345) stored frozen at -20 degrees C in their original containers (leakproof 100 mL plastic bottles) upon retesting for ethanol resulted in concentrations that were significantly lower (average loss = approximately 30%) than those prior to their storage at -20 degrees C (p < or = 0.0001). The observed loss of ethanol was independent of the method of thawing or the concentration of ethanol in the samples, but was dependent on the sample volume in the container, i.e., the larger the volume of sample the larger the magnitude of ethanol loss. The loss of ethanol was determined to be due to improper sealing by a Styrofoam integrity seal attached to the mouth of the container. Accordingly, adopting leakproof plastic containers that do not contain Styrofoam integrity seals, but rather an outside and across the cap tape integrity seal for evidence collection and long-term storage, will prevent loss of ethanol due to evaporation. PMID- 12762547 TI - Zaleplon and driving impairment. AB - Zaleplon, a sedative-hypnotic, was identified in the blood of a subject arrested for impaired driving. Symptoms reported were those of central nervous system (CNS) depression. The zaleplon concentration was determined to be 0.13 microg/mL. Symptoms included slow movements and reactions, poor coordination, and lack of balance. Although no quantitative relationship between blood concentrations and degree of driving impairment is currently possible, it is reasonable to conclude that because of its specific activity as a sedative-hypnotic, blood concentrations consistent with doses exceeding therapeutic concentrations of zaleplon have the potential to cause impairment of psychomotor function, and would impair a person's level of consciousness and driving ability. PMID- 12762548 TI - The analysis of an intracerebral hematoma for drugs of abuse. AB - Toxicological investigations were performed on an intracerebral hematoma, antemortem blood, and postmortem blood of an individual who was found unresponsive in his home. The hematoma was found to have ethanol at a concentration of 0.05% (w/v), and benzoylecgonine (a cocaine metabolite) was also confirmed at a concentration of 0.43 mg/L by specific analysis using gas chromatography/mass spectrometry (GC/MS). These results enabled the pathologist to record the cause of death as intracerebral hemorrhage due to acute cocaine intoxication. PMID- 12762549 TI - A fatal drug interaction between oxycodone and clonazepam. AB - A case is presented of a fatal drug interaction caused by ingestion of oxycodone (Oxycontin) and clonazepam (Klonapin). Oxycodone is an opium alkaloid used in long-term pain management therapy. Clonazepam is a benzodiazepine used for the treatment of seizures and panic disorders. The Drug Abuse Warning Network (DAWN) has reported an increase of 108% in the last two years of emergency department episodes related to Oxycontin. Six billion prescriptions were written for Oxycontin in the year 2000, an 18-fold increase from four years previous (1). Oxycontin has recently gained enormous notoriety at the local and national levels; however, there are very few previously documented cases of lethal drug interactions between oxycodone and clonazepam. Synergistic effects between these two drugs are postulated to arise from different agonistic mechanisms producing similar physiological changes. It is also theorized that clonazepam may inhibit the metabolism of oxycodone. A 38-year-old white female was found dead in Jefferson County, Tennessee in March of 2001. The deceased had physical evidence of previous drug abuse and positive serological findings of hepatitis B and C. Prescription pill bottles filled under the name of the deceased, as well as another name, were found with the body. Serum, urine and gastric contents from the deceased were screened for numerous drugs and metabolites using a combination of thin layer chromatography and immunoassay techniques (EMIT and FPIA). Analysis of biological specimens from the deceased revealed the presence of: benzodiazepines, opiates (oxycodone), and trazodone metabolites in the serum; cannabinoids, benzodiazepines, opiates (oxycodone), trazodone, trazodone metabolites, nicotine, and nicotine metabolite in the urine; and benzodiazepines, opiates (oxycodone), nicotine, and nicotine metabolite in the gastric contents. Quantitative analyses for clonazepam was performed by high performance liquid chromatography (HPLC) and revealed a plasma concentration of 1.41 microg/mL. Plasma oxycodone and urine 11-nor-carboxy-delta-9-tetrahydrocannabinol concentrations were determined by gas chromatography/mass spectrometry and revealed concentrations of 0.60 microg/mL and 27.9 ng/mL, respectively. The deceased had pathologies consistent with severe central nervous system (CNS) and respiratory depression produced by high concentrations of clonazepam and oxycodone including collapsed lungs, aspirated mucus, and heart failure. The pathologies were sufficient to cause death, which was officially attributed to a drug overdose; however, the manner of death was unknown. PMID- 12762550 TI - Genetic variation for 7 STR loci in a Colombian population (Department of Valle del Cauca). PMID- 12762551 TI - Two X-chromosome STR loci DXS6807 and DXS7133 frequency data in Chinese population. PMID- 12762552 TI - Distribution of D5S818, D7S820, D8S1179, D13S317, D18S51 alleles in a central Italian population sample. PMID- 12762553 TI - Forensic biology. PMID- 12762554 TI - Commentary on: Platt MS, Kohler LJ, Ruiz R, Cohle SD, Ravichandran P. Deaths associated with liposuction: case reports and review of the literature. J Forensic Sci 2002;47(1): 205-207. PMID- 12762555 TI - Approaching 100 years of medical and university education in Singapore. PMID- 12762556 TI - Medicine in stamps. Paracelsus (1493-1541): the man who dared. PMID- 12762557 TI - Comparison of four point-of-care HbA1c analytical systems against central laboratory analysis. AB - OBJECTIVE: To assess four commercially available point-of-care HbA1c analytical systems with respect to (1) inaccuracy against the central laboratory HbA1c method and (2) imprecision against the HbA1c analytical goal of coefficient of variation < 3%. METHODS: Analytical inaccuracy was assessed by analysis of 110 patient samples on all five analytical platforms (Biorad Diastat, Drew DS5, Bayer DCA 2000, Nycomed Nycocard and Roche Tinaquant (used in central lab)). Analytical imprecision was assessed by analysis of two levels of patient sample four times daily for six days, as well as analysis of two levels of commercial control. RESULTS: Deming linear regression for agreement: Diastat=0.98 x Tinaquant + 0.36; DS5=1.23 x Tinaquant - 0.65; DCA2000=0.95 x Tinaquant + 0.63; Nycocard=0.94 x Tinaquant + 0.92. Analytical coefficients of variation (CVs) at Tinaquant HbA1c levels of 6.2-10.8% were: Tinaquant 0.8-1.1%, Diastat 1.6-6.6%, DCA2000 2.6-7.2%, DS5 5.1-11.7%, Nycocard 8.5-15.3%. Two HbE samples gave elevated HbA1c results with the DS5 method. CONCLUSIONS: The Diastat and DCA2000 systems gave the best performance with acceptable imprecision and good agreement with both the central lab and each other. The DS5 was less precise with a significant positive bias compared to the other methods and interference from HbE, while the Nycocard system showed the poorest precision in the evaluation. The Diastat and DCA2000 systems appear to be satisfactory analytical alternatives to both central laboratory (Tinaquant) testing and each other. PMID- 12762558 TI - A review of 2,517 childhood injuries seen in a Singapore emergency department in 1999--mechanisms and injury prevention suggestions. AB - BACKGROUND: Childhood injuries cause significant mortality and morbidity in Singapore. With injury surveillance, patterns of repeated injury can be identified and injury prevention strategies devised. METHODS: We conducted a retrospective study of all children aged 12 and below seen for trauma in an Emergency Department over one year. Data captured in the real-time computer system was studied with regards to patient profile, mechanism of injury and patient disposition. Clinical summaries were extracted with follow-up telephone interviews done. RESULTS: Two thousand five hundred and seventeen children aged 12 and below were seen for accidental trauma in 1999, accounting for 37.1% of the total attendance for that age. Mean age was 7.7 years with males making up 62.7%. Home injuries (56.4%) were the most common, followed by road-related (14.4%), sports (8.2%) and playground injuries (7.4%). 48.5% sustained head and face injuries. Pre-school children (age <5) were more likely to sustain home injuries (p<0.0001), a higher proportion of head injuries (p<0.0001), foreign bodies, burns and poisoning compared to school-going children (age 6-12), who were more likely to sustain injuries in road accidents, sports, at playgrounds or schools, with more limb, trunk and multi-trauma. We highlight drownings, falls from height, rollover falls from beds, slamming door injuries, the low use of child car restraints, bicycle injuries and playground falls as areas of concern. CONCLUSION: Several injury prevention strategies have been suggested and it is hoped these may contribute to addressing preventable childhood injuries in Singapore. We also advocate the establishment of a national childhood injury surveillance database. PMID- 12762559 TI - Erectile dysfunction in Singapore: prevalence and its associated factors--a population-based study. AB - PURPOSE: To study the prevalence of erectile dysfunction (ED) in Singapore males aged 30 and above and its association with demographic, medical and other risk factors. METHODOLOGY: A population based cross sectional study of 729 men aged 30 and above in Singapore was conducted using the abridged, five-item version of the International Index of Erectile Function (IIEF-5). Presence of erectile dysfunction was defined as IIEF-5 score of less than 21. Erectile dysfunction was further categorised into mild (IIEF-5: 16-20), moderate (IIEF-5: 11-15) and severe (IIEF-5: <11). A logistic regression model was used to identify significant independent risk factors for ED. RESULTS: Overall, 51.3% of respondents (n=374) reported some degree of erectile dysfunction. Of these, 23.2% have mild ED, 8.8% have moderate ED and 19.3% had severe ED. The prevalence of ED increased from 42.8% for men in their forties to 77.4% in their sixties. The prevalence of severe ED increased from 9.1% in men in their forties to 43.5% in their sixties and 77.0% in those aged 70 and above. Age above 50 years is the single most significant risk factor on multivariate analysis when adjusted for all confounding factors. Other important risk factors include Indian ethnic group, lower household income, physical inactivity, diabetes mellitus and cardiac diseases. CONCLUSION: Erectile dysfunction is common amongst Singaporean men. Age is the single most important physiologic factor affecting erectile function. The prevalence and severity increased significantly with age after 50 years old. With an ageing population, erectile dysfunction may become a significant health problem. Health care providers should plan their resources accordingly. PMID- 12762560 TI - A case of Bizarre Parosteal Osteochondromatous Proliferation of the hand. AB - We report a case of Bizarre Parosteal Osteochondromatous Proliferation (BPOP) of the hand that was initially misdiagnosed as an osteochondroma. BPOP, although a benign lesion, behaves aggressively with rapid growth and high risk of local recurrence after local resection. The distinguishing features of BPOP as opposed to an osteochondroma are discussed. PMID- 12762561 TI - Ketamine associated psychedelic effects and dependence. AB - Ketamine, a dissociative anaesthetic in use since 1970, produces prominent psychoactive effects in humans. Its non-medical use has raised concerns in many countries, including Singapore. This paper narrates the psychedelic and psychotic effects of ketamine in two ketamine dependent patients who have presented to the psychiatric service. These effects were dose-related and comprised multimodal hallucinatory experiences, a sense of slowing, paranoid ideation and enhancement of sexual, musical and sensory enjoyment. In both ketamine users the psychotic symptoms resolved quickly with symptom-targeted treatment. However, breaking the ongoing addiction cycle seemed more difficult. The neuro-pharmacological mechanisms of these phenomena are largely due to its complex multi-receptors actions, notably through the excitatory amino acids through mainly the N-methy-D aspartate (NMDA) receptors. The detection of ketamine abuse requires a high index of suspicion and needs to be considered when there is an acute presentation with multi-modal hallucinations and psychosis. PMID- 12762562 TI - Intra-arterial embolisation in otolaryngology--a four-year review. AB - This is a retrospective review of otolaryngology patients who underwent intra arterial embolisation in a tertiary hospital over a four-year period. Thirty patients were reviewed. Seven (23.3%) were embolised electively before resection of hypervascular head and neck tumours. Of the emergency indications, nine patients (30%) had intractable epistaxis from non-oncologic causes, 14 (46.7%) had bleeding of oncologic origin from the head and neck. In all, six patients required further procedures within the first week, 2 (6.7%) suffered permanent cranial nerve deficits. Our results show that intra-arterial embolisation is both useful and safe in elective and emergency settings in otolaryngology, with complication rates comparable to previous publications. The survival of oncology patients with otherwise fatal haemorrhage was significantly prolonged. PMID- 12762563 TI - MR appearance of interpeduncular lipoma. AB - Interpeduncular lipoma (IPL) is a very rare benign intracranial lesion. Non invasive diagnosis of this condition is important, as these lesions are usually asymptomatic and are found incidentally. We describe a rare case of IPL in a 35 year-old woman presenting with headache of long duration and a recent increase in severity and frequency of headache. There was no neurological abnormality on clinical examination. MR imaging demonstrated a homogeneous lobulated T1 hyperintense and T2-hypointense lesion in interpeduncular fossa. A T1-weighted fat suppression sequence with chemical shift method confirmed the fatty nature of the lesion. The patient was managed conservatively. PMID- 12762565 TI - Clinics in diagnostic imaging (81). Hypothalamic glioma with diencephalic syndrome. AB - A 15-month-old boy presented with failure to thrive. Physical examination revealed third degree malnutrition. MR imaging showed a large lobulated mass in the suprasellar region. The mass was hypointense on T1-weighted MR images, hyperintense on proton density-weighted and T2-weighted MR images, and enhanced homogeneously. The patient underwent a craniotomy for a subtotal resection of the tumour. Histopathological examination revealed a juvenile pilocytic astrocytoma that caused the diencephalic syndrome. Clinical presentation and imaging findings of the various tumours or diseases that can cause suprasellar masses in children are discussed and illustrated via further examples. PMID- 12762564 TI - A case of cutaneous leukocytoclastic vasculitis associated with gabapentin. AB - Gabapentin was approved for use in the USA in 1994 as an add-on drug in adults who have partial seizures either alone or with secondary generalised seizures. Its use has been expanded to include treatment for other conditions such as neuropathic pain and paraesthesiae. Gabapentin was prescribed for our patient who had persistent left-sided hemi-paraesthesiae consequential of a previous thalamic infarct. One week after commencement of gabapentin, she developed an adverse reaction in the form of a purpuric rash over bilateral lower limbs. Skin biopsy revealed histological features of cutaneous leukocytoclastic vasculitis. This rash completely resolved with withdrawal of gabapentin and steroidal treatment. This cutaneous adverse reaction to gabapentin has not been reported in the medical literature. PMID- 12762566 TI - Air pollution effects in the elderly: introduction. PMID- 12762567 TI - Epidemiological aspects and early detection of chronic obstructive airway diseases in the elderly. AB - The burdens of chronic obstructive airway diseases among the elderly in Europe, and worldwide, are increasing. Although asthma is common in all ages, the main airway disease affecting the elderly is chronic obstructive pulmonary disease (COPD). The aim of this paper is to review the prevalence and incidence of COPD on the basis of population studies. As the prevalence estimates of asthma are probably well known, only the incidence and remission of asthma will be discussed. The underdiagnosis of obstructive airway diseases is huge. A Dutch programme for early detection of obstructive airway disease among the elderly has, thus, been included in the presentation. A prerequisite for fighting COPD is to acquire data on illnesses and death. COPD has only recently been defined by cut-off points of spirometric outcomes, which is why measures of the prevalence of COPD have been distorted by use of a large number of different diagnostic terms and lung function criteria. The prevalence of clinically-relevant COPD has been estimated in several community studies to 4-6% in adult population samples, with a considerable increase by age, particularly among smokers. The incidence of COPD not only increases heavily with age and smoking, but also occupational exposure to dust, gas and damp. Precise estimates of the incidence of COPD or spirometric airflow limitation are not available. Demographic changes will result in a further substantial increase of chronic obstructive airway disorders, mainly chronic obstructive pulmonary disease, among the elderly. The increasing burden of chronic obstructive pulmonary disease has to come to the awareness of the public, governments, health authorities, and industry. PMID- 12762568 TI - Determinants of prognosis of COPD in the elderly: mucus hypersecretion, infections, cardiovascular comorbidity. AB - In this paper, the authors update the present knowledge about three risk factors for the prognosis of chronic obstructive pulmonary disease (COPD), which may be particularly relevant in elderly people: mucus hypersecretion, respiratory infections, and cardiovascular comorbidity. Chronic mucus hypersecretion (CMH) is a common respiratory symptom in old age, the relevance of which is analysed on the basis of data collected during the first three rounds of the Copenhagen City Heart Study. In subjects aged > or = 65 yrs, CMH was a strong predictor of the incidence of respiratory infections in a 10-yr follow-up period and it was also a strong predictor of death from COPD (relative risk=2.5). However, CMH was associated with consistently lower forced expiratory volume in one second (FEV1) values, but not with an accelerated decline of FEV1 in this sample of an elderly population. Acute respiratory infections (ARI) are extremely common at all ages, mostly mild self-limiting illnesses at a young age, but severe often fatal illnesses in elderly people already affected by a chronic disease such as COPD. This paper summarises the present knowledge about aetiology, pathology, prognostic relevance, and prevention of ARI. Furthermore, the areas in which further research is needed are listed. Clinical cohort studies clearly support the relevance of cardiovascular comorbidity for the short-and long-term prognosis of elderly subjects affected by severe COPD. In this paper, the recently demonstrated association between particulate air pollution and cardiovascular events is reported to suggest the presence of an extremely susceptible cluster of elderly subjects in the population identified by the copresence of chronic obstructive pulmonary disease and cardiovascular comorbidity. PMID- 12762569 TI - Indoor air pollution and respiratory health in the elderly. AB - People spend about > or = 80-90%, of their daily time indoors, elderly people especially at home. Thus, it is important to investigate possible health effects of indoor air pollutants and to consider their contributions to the total human exposure. This report summarises current knowledge on health effects of three common indoor air pollutants, respirable suspended particles, nitrogen dioxide and environmental tobacco smoke, with focus on the adults and the elderly. Preliminary findings on exposure distributions and health effects of these pollutants in older subjects of two panel studies carried out in Italian general populations will also be reported. The two indoor pollution studies were performed in the Po Delta area in North Italy (428 subjects and 140 houses investigated) and in Pisa in Central Italy (761 subjects and 282 houses investigated). Individuals aged > or = 65 yrs spent a significantly larger number of hours at home than the other age groups both in winter and in summer. A trend of higher occurrence of acute respiratory symptoms in the presence of environmental tobacco smoke was shown in comparison to the unexposed elderly both in winter (31 versus 29%) and summer (33 versus 16%). The occurrence of acute respiratory symptoms was consistently higher in relation to the high respirable suspended particles-index exposure compared to low exposure (33 versus 27%, in winter, 27 versus 21% in summer). Both the presence of environmental tobacco smoke at home and exposure to the high respirable suspended particles-index were associated with a decrease in the mean daily peak expiratory flow. PMID- 12762570 TI - Questionnaires, spirometry and PEF monitoring in epidemiological studies on elderly respiratory patients. AB - Questionnaires are the most used subjective instrument of measurement in respiratory epidemiology. The standardisation of the questionnaires aims to limit bias by maximising validity and reliability, and comparability. Within the European Union project BIOMED1, a compendium of respiratory standard questionnaires (CORSQ) was developed for adults covering 18 topics from general information to early life events, through environmental risk factors and respiratory symptoms and diseases. Reliable spirometry data needs a rigorous quality control programme, as in the "Salute Respiratoria nell'Anziano" (Sa.R.A.) project, Italian for "Respiratory Health in the Elderly". Reproducibility rates were 95.8% for forced expiratory volume in one second (FEV1). Male sex and age were independent risk factors for a poorer reproducibility, as well as cognitive and physical impairment (shorter 6-min walking distance) and lower educational level for a poorer acceptability. Reference values for people aged 65-85 yrs have been produced; these results suggest that the effect of aging should be corrected for physical and mental disability. A revision of interpretative strategies included in current guidelines is needed. Peak expiratory flow monitoring has several methodological problems: reliability and sensitivity of the measurement in order to detect changes in airway calibre; compliance with long-term monitoring; choice of the best variability index; difference between asthmatic and nonasthmatic subjects; age-related differences. Despite these methodological problems, peak expiratory flow monitoring has been successfully used in the evaluation of the effects of air pollution in normal and asthmatic subjects, and in the elderly. PMID- 12762571 TI - Short-term effects of ambient particles on mortality in the elderly: results from 28 cities in the APHEA2 project. AB - Within the framework of the APHEA2 (Air Pollution on Health: a European Approach) project, the effects of ambient particles on mortality among persons > or = 65 yrs were investigated. Daily measurements for particles with a 50% cut-off aerodynamic diameter of 10 microm (PM10) and black smoke (BS), as well as the daily number of deaths among persons > or = 65 yrs of age, from 29 European cities, have been collected. Data on other pollutants and meteorological variables, to adjust for confounding effects and data on city characteristics, to investigate potential effect modification, were also recorded. For individual city analysis, generalised additive models extending Poisson regression, using a locally weighted regression (LOESS) smoother to control for seasonal effects, were applied. To combine individual city results and explore effect modification, second stage regression models were applied. The per cent increase (95% confidence intervals), associated with a 10 microg x m(-3) increase in PM10, in the elderly daily number of deaths was 0.8%, (0.7-0.9%) and the corresponding number for BS was 0.6%, (0.5-0.8%). The effect size was modified by the long-term average levels of nitrogen dioxide (higher levels were associated with larger effects), temperature (larger effects were observed in warmer countries), and by the proportion of the elderly in each city (a larger proportion was associated with higher effects). These results indicate that ambient particles have effects on mortality among the elderly, with relative risks comparable or slightly higher than those observed for total mortality and similar effect modification patterns. The effects among the older persons are of particular importance, since the attributable number of events will be much larger, compared to the number of deaths among the younger population. PMID- 12762572 TI - Air pollution and mortality in The Netherlands: are the elderly more at risk? AB - The association between daily mortality and short-term variations in the ambient levels of ozone (O3), black smoke (BS), sulphur dioxide (SO2), nitrogen dioxide (NO2), carbon monoxide (CO) and particulate matter was studied in The Netherlands. Daily total and cause-specific mortality counts (cardiovascular, chronic obstructive pulmonary disease (COPD) and pneumonia), air quality, temperature, relative humidity and influenza data were obtained from 1986-1994. The relationship between daily mortality and air pollution was modelled using Poisson regression analysis. All pollution mortality associations were adjusted for potential confounding due to long-term trends, seasonal trends, influenza epidemics, ambient temperature, ambient relative humidity, day of the week and holidays, using generalised additive models. Statistically significant associations were mostly found in the elderly, that is the age categories of 65 74 and > or = 75 yrs for the pollutants PM10 (particles with a 50%, cut-off aerodynamic diameter of 10 microm), BS, SO2, NO2 and CO. This may partly be due to a better precision of relative risk (RR) estimates for the larger numbers of deaths in these age groups. Significant associations for those < 65 yrs were found for O3 (total and COPD mortality), PM10 (pneumonia), NO2 (pneumonia) and CO (pneumonia). RR estimates for deaths between 45-65 yrs tended to be smaller than those in > 65 yrs, with the exception of ozone; for cardiovascular mortality the RR for PM10, O3 and CO were similar in these age groups. In conclusion, larger relative risks for air pollution were mostly found in the elderly except for ozone and for death-cause pneumonia which showed larger relative risk in younger age groups. PMID- 12762573 TI - Particulate air pollution and hospital admissions for cardiorespiratory diseases: are the elderly at greater risk? AB - A systematic literature review suggests that particulate air pollution is associated with daily admissions for both respiratory and cardiac diseases in people aged > 65 yrs. A model of acute effects is proposed which shows how admissions can be brought forward by a relatively short period of time as well as events being added that would not have happened at all except for air pollution. A model of the effects of air pollution on chronic disease is proposed that provides the background of long-term vulnerability upon which the increased short term vulnerability is superimposed. A study of daily hospital admissions in London shows that for respiratory disease the relative risks of admission associated with particles reduce with increasing age, while for cardiac disease, there is no trend. When the attributable risk is estimated using baseline admission rates for respiratory disease, it is children who have the highest attributable risk, followed by the elderly. For cardiac disease there is a steep increase in attributable risk with age, reflecting the dominant influence of baseline risks. The attributable risk for cardiovascular disease in the elderly is considerably greater than for respiratory disease, due to higher baseline admission rates. PMID- 12762574 TI - Mechanism of lung injury caused by PM10 and ultrafine particles with special reference to COPD. AB - Particulate air pollution (particles with a 50% cut-off aerodynamic diameter of 10 microm (PM10)) has strong associations with the adverse health events detected by epidemiological studies in chronic obstructive pulmonary disease patients in diverse geographical locations. Transition metals may determine the toxic effects of PM10 through oxidative stress which may be injurious as shown by an increase in airspace epithelial permeability and may lead to inflammation through the activation of transcription factors for pro-inflammatory genes in both macrophages and epithelial cells. Recently, the present authors have shown that particulate air pollution may cause further molecular events that enhance transcription factor activation by causing acetylation of histones leading to unwinding of deoxyribonucleic acid (DNA) enhancing transcription factor DNA binding and increasing transcription for pro-inflammatory genes. Further, ultrafine particles (< 100 nm diameter), which have marked toxicity, may be responsible for some of the PM10 adverse effects. The current authors have demonstrated that ultrafine carbon black (ufCB) does not have its effects via transition metal-mediated mechanisms. However, ufCB and other ultrafines generate free radicals at their surface as measured by a number of chemical assays and are able to cause oxidative stress to cells and this is likely to be a factor in their ability to cause inflammation. Changes in calcium resulting from oxidative stress within cells may be an additional factor leading to transcription of pro inflammatory genes. Understanding the mechanisms of the harmful effects of particulate air pollution in chronic obstructive pulmonary disease may help in risk strategy for individuals who are susceptible to the effects of air pollution. PMID- 12762575 TI - Genetic aspects of susceptibility to air pollution. AB - Inter-individual variation in human responses to air pollutants suggests that some subpopulations are at increased risk, and it is increasingly clear that genetic background is an important susceptibility factor. Genetically standardised animal models provide useful investigative tools. Linkage analyses using inbred mice identified chromosomal segments (quantitative trait loci (QTL)), with genes controlling susceptibility to the lung inflammatory (chromosome 17), injury (chromosome 11), and hyperpermeability (chromosome 4) responses to ozone (O3) exposure. An immune dysfunction response induced by exposure to sulphate-associated particles is linked to the identical chromosome 17 and 11 QTLs described for O3 susceptibility, thus similar genetic mechanisms may be controlling pulmonary responses to these pollutants. Candidate genes within the QTLs on chromosomes 4 and 17 include the toll-like receptor 4 and the pro-inflammatory cytokine, tumour necrosis factor-alpha, respectively. Functional analyses strongly support a role for these candidate genes in determining susceptibility to O3 and particulates. Because striking linkage homology exists between the human and mouse genomes, candidate susceptibility genes identified in the mouse are likely to aid research aimed at understanding human genetic factors that contribute to differential susceptibility. To date, no studies have examined the interaction between age and genetic background in the development of air pollution-induced lung disease. However, investigations have suggested an influence of age on genetic susceptibility to lung cancer and other diseases, which indicate that an interaction between age and genetic background may be important in air pollution disease pathogenesis. PMID- 12762576 TI - Subpopulations at increased risk of adverse health outcomes from air pollution. AB - Epidemiological research to identify subpopulations with enhanced susceptibility to air pollution is still at an early stage. From the available studies, there is evidence that both "endogenous" and "exogenous" factors contribute to individual susceptibility. Females and the elderly are at an increased risk of pollution related diseases. Moreover, some chronic clinical conditions seem to be good candidates for identifying the "frail" populations: chronic obstructive pulmonary disease including asthma, coronary heart diseases, congestive heart failure, and heart rhythm disorders. It seems clear that epidemiological research on susceptibility in the future should investigate the underlying biological and physiological mechanisms, in addition to the environmental and toxicological effects. PMID- 12762577 TI - Nonallergic respiratory morbidity improved along with a decline of traditional air pollution levels: a review. AB - The tremendous decline of combustion-derived emissions of traditional air pollutants such as sulphur dioxide (SO2) and total suspended particles (TSP) in Eastern Germany shortly after German reunification in 1990 provided the unique opportunity to study trends of prevalence of respiratory illness along with the improvement of air quality. The present review focused on the results of two repeated surveys of nonallergic respiratory illness in children living in East Germany. The crude prevalence of respiratory illness such as lifetime bronchitis, otitis media, tonsillitis, frequent colds, and frequent cough decreased during the 1990s in East German children. For two surveys the effect estimates showed consistently statistically significant association of TSP with bronchitis. For SO2 statistically significant associations with frequent colds were also consistently found, whereas at least one survey reported higher statistically significant effect estimates for tonsillitis, otitis media, frequent cough and reduced forced vital capacity. The present study concludes that the prevalence of nonallergic respiratory illness in East German children is associated with sulphur dioxide and total suspended particles and that the improvement of air quality has beneficial effects on respiratory health. PMID- 12762578 TI - Air pollution and the elderly: oxidant/antioxidant issues worth consideration. AB - The elderly are frequently classified as being particularly susceptible to air pollution. The basis of this increased sensitivity is not known but it is likely that it is linked to age-related impaired function of the lung. However, given this uncertainty and increased impact of air pollution of this section of the community there is a need to explore the mechanisms involved. Gaseous air pollutants such as ozone and nitrogen dioxide, or many of the components adsorbed onto the surface of respirable particles, are either powerful oxidants or capable of generating free radicals. If unabated, oxidants can cause injury to the delicate cells that line the respiratory tract. Small molecular weight antioxidant defences present in respiratory tract lining fluid (RTLF) represent the first line of defence against a range of oxidants that enter the lung. The quantity and quality of the RTLF airways antioxidant network are, therefore, likely to be important determinants of the impact of the oxidant challenge on the underlying respiratory epithelium. As yet, comprehensive information on the distribution and variability of respiratory tract lining fluid antioxidant defences is only available in young, mainly healthy volunteers. Studies undertaken in patients with a range of respiratory diseases suggest that marked changes in the distribution of respiratory tract lining fluid antioxidants can occur. Information is not currently available on the impact of ageing on the respiratory tract lining fluid antioxidant defence network. As several respiratory tract lining fluid antioxidants are of dietary origin, the elderly, who often have different dietary patterns to younger individuals, may have decreased availability of important antioxidants. Given these possibilities, a better understanding of respiratory tract lining fluid antioxidant defences in the aged lung is warranted. PMID- 12762579 TI - Elderly humans exposed to concentrated air pollution particles have decreased heart rate variability. AB - Air pollution particles are thought to kill > 500,000 people worldwide each year. The population most at risk appears to be elderly people with respiratory and cardiovascular disease. As yet, no commonly accepted mechanism has been proposed which can explain the cause of these deaths. Heart rate variability (HRV) was assessed in healthy elderly adults between the ages of 60 and 80 who were exposed twice for 2 h: once to clean air and once to concentrated ambient air pollution particles (CAPS). Changes in HRV were measured immediately before, immediately following, and 24 h after exposure. Elderly subjects experienced significant decreases in HRV in both time and frequency domains immediately following exposure. Some of these changes persisted for at least 24 h. These data were compared with HRV data collected from young healthy volunteers exposed to CAPS in a previous study, in which no CAPS-induced changes in HRV were found. These concentrated ambient air pollution particle-induced changes in heart rate variability in a controlled human exposure study extend similar findings reported in recent panel studies and suggest potential mechanisms by which particulate matter may induce adverse cardiovascular events. PMID- 12762580 TI - Case-crossover design in air pollution epidemiology. AB - The case-crossover design was developed to study the effects of transient, short term exposures on the risk of acute events, such as myocardial infarction, in the early 1990s. This paper illustrates how the principles of case-crossover design are related to the principles of crossover and case-control designs and stipulates the possibilities of case-crossover design in air pollution epidemiology. In the case-crossover design, the study population consists of subjects who have experienced an episode of the health outcome of interest. Similar to a crossover study, each subject serves as his or her own control. As in a matched case-control study, the inference is based on a comparison of exposure distribution rather than the risk of disease. The case-crossover study is most suitable for studying relations with the following characteristics: 1) the individual exposure varies within short time intervals; 2) the disease has abrupt onset and short latency for detection; and 3) the induction period is short. Case-crossover design allows use of routinely monitored air pollution information and at the same time makes it possible to study individuals rather than days as the unit of observation. Case-crossover design is amenable for studying the effects of varying short-term air pollution exposure on health outcomes with an abrupt onset, such as myocardial infarction or asthma attack. PMID- 12762581 TI - Health impact assessment of air pollution: providing further evidence for public health action. AB - Many epidemiological studies have demonstrated the importance of air pollution as a risk factor and characterised dose-response relationships between health endpoints and pollutants. The association between particulate matter (PM) and health is generally regarded as causal, and a nonthreshold linear relationship with, for example, mortality and hospital admission has been observed in several settings. The ubiquitous PM air pollution is likely to have a large overall impact on human health, even if risks are relatively small. There have recently been a large number of papers reporting quantitative estimations of the health impact of PM on health, as measured by the proportion of excess events that are attributable to PM exposures in the general population, mainly in industrialised countries. For example, in the eight largest Italian cities it has been estimated that concentrations beyond 30 microg x m(-3) are responsible for about 3,500 extra deaths per year. A similar study has been carried out for France, Austria and Switzerland. These evaluations fill a knowledge gap between the laboratory and clinical studies on the pathophysiological mechanisms, the epidemiological research on the nature and strength of the association at the population level, and the risk management needs for developing appropriate preventive policies. Some limitations in the methodology deserve further research, however health impact assessment type studies are informative and effective tools of communication with the general public and policy makers. PMID- 12762582 TI - The need for a focus on air pollution research in the elderly. AB - During recent years an increasing focus has been directed towards the adverse health effects associated with ambient air pollution. Elderly people appear to be particularly susceptible to the adverse effects involving the respiratory and cardiovascular systems, resulting in symptoms, exacerbations of disease and even mortality. From an epidemiological point of view it is essential to obtain a more detailed description and identification of factors associated with these health effects. Novel study designs are needed with complementary exposure and biomedical characterisation. Long-term prospective studies are required. A better understanding of the pathophysiological mechanisms is considered important and requires an interaction between epidemiological and mechanistic studies in elderly individuals with or without complementary diseases that put in them especially at risk. Generally a synergy between complementary disciplines is warranted to move this important research area forward, also including in vitro models of cell responses in the elderly, animal models of diseases of the elderly, together with controlled air pollution exposure studies identifying health-related events and mechanisms. The generation of an understanding of air pollution effects in the elderly, at an elevated level, is a prerequisite to substantially reducing the adverse health effects of this population group. At local, national and European Union levels, some steps have been taken to support the research in this area. A major focus on the adverse air pollution effects in the elderly requires a long-term commitment that still remains to be established. PMID- 12762583 TI - The dynamics of morphological measures and lipid peroxidation during aging of the cerebral cortex in humans. PMID- 12762584 TI - Structure of the insular region of the rat neocortex. PMID- 12762585 TI - Instrumentalization of movements evoked by stimulation of the motor cortex by food reinforcement in dogs. AB - The possibility that hindlimb movements (elevations) evoked by stimulation of the corresponding contralateral area of the motor cortex could be instrumentalized by reinforcement with food was demonstrated, contradicting some previously published data. Operant movements (interstimulus voluntary high elevations of the hindlimb) were acquired as a result of consistent combinations: cortical stimulation - movement - food. Acquisition required more than 50-200 combinations. Delivery of food was accompanied by a click at exactly the moment at which the hindlimb reached the required height. The click became the food-related conditioned signal and served as a secondary operant reinforcement, which facilitated acquisition of the operant movement. These results support the view that the motor cortex can have an immediate role in forming "operant" temporary connections (motivation movement) and that simple operant movements can be initiated via this arc. PMID- 12762586 TI - Experimental analysis of the processes of systems genesis: expression of the c fos gene in the chick brain during treatments inducing the development of the species-specific results-of-action acceptor. AB - The aim of the present work was to identify which parts of the chick brain are activated during treatments inducing the development of the preference to follow a species-specific object (a stuffed chick). Expression of the gene for the transcription factor c-fos was used as the molecular marker for neuron activation. Day-old chicks lacking visual experience were placed for 90 min in a freely rotating squirrel wheel or were subjected to stimulation with a loud noise for 180 min. The animals' preference was tested 24 h after stimulation ended. Both types of stimulation induced the formation of a marked preference to follow the "natural" object in the chicks. c-fos expression was analyzed in sections from the brains of chicks stimulated for 45 min. Increases in the quantity of c fos mRNA were seen after each type of stimulation in the medial part of the caudal neostriatum. In addition, stimulation in the squirrel wheel was accompanied by high levels of c-fos expression in the paraolfactory lobes, while sound stimulation gave high levels of c-fos expression in the ventral and caudal parts of the archistriatum. These structures of the chick brain are of great interest for studies of the cellular and molecular mechanisms of the formation of the species-specific results-of-action acceptor in the mother-following functional system. PMID- 12762587 TI - Differences in serotonin and dopamine metabolism in the rat brain in latent inhibition. AB - Monoamine oxidase activity during deamination of serotonin and dopamine in the amygdaloid complex, hippocampus, striatum and prefrontal cortex of the brain was studied at different stages in the formation of latent inhibition, acquired by rats on the basis of a conditioned passive avoidance reaction. Latent inhibition is a behavioral phenomenon consisting of a worsening in learning using a stimulus when attention to this stimulus is quenched before acquisition of the conditioned response. At the stage of preliminary repeated presentation of the conditioned stimulus, serotonin metabolism was seen to change in the amygdaloid complex and striatum, while dopamine metabolism changed in the amygdaloid complex and hippocampus. Metabolic shifts in transmitter handling during quenching of attention to the stimulus occured in the opposite directions: the serotonin deaminating activity of monoamine oxidase increased, while the dopamine deaminating activity of the enzyme decreased. Unlike the reaction to the quenched stimulus, the latent inhibition effect, seen on testing the conditioned response after conditioning of the pre-exposed stimulus with unconditioned reinforcement, was accompanied by changes in serotoninergic activity only. High levels of serotonin deamination by monoamine oxidase were seen in the amygdaloid complex and striatum. In addition, there was a reduction in serotonin deamination in the prefrontal cortex specific for the stage of testing latent inhibition. At the same time, dopamine metabolism did not change in any of the brain structures studied in latent inhibition. These data lead to the conclusion that the latent inhibition effect may be based on increases in serotoninergic activity in subcortical brain structures--the amygdaloid complex and the striatum--induced by reactions to presentation of the quenched stimulus. PMID- 12762588 TI - Analysis of the linkage of the Taq1A and Taq1B loci of the dopamine D2 receptor gene with schizophrenia in patients and their siblings. AB - The linkage of the polymorphous markers Taq1A and Taq1B of the DRD2 dopamine receptor gene, located in region 11q22-11q23 of chromosome 11, with schizophrenia was studied. The investigation involved 29 complete families containing concordant and discordant sibling pairs. Common alleles at locus Taq1A were found significantly more frequently in concordant pairs (p = 0.04), and there was a tendency to a higher frequency of transmission of the maternal allele as compared with the paternal allele (p = 0.06). No such relationships were seen in the case of the Taq1B locus. Neither locus showed any significant difference in the frequency of common alleles in discordant pairs or in the predominance of allele transmission from one of the parents. These data demonstrate a possible linkage with schizophrenia for the Taq1A marker but not for the Taq1B marker. PMID- 12762589 TI - Role of the dopaminergic system of the brain in the effects of glucocorticoid hormones. AB - The conditioned reinforcement properties of the synthetic glucocorticoid dexamethasone and the possible mechanisms of its action were studied using a conditioned place preference response. On test day 1, male Wistar rats were placed in a two-chamber apparatus and the time spent in each sector was measured for 10 min. Over the next six days, combinations of one of the chambers with administration of agents (reinforcement) were presented using injections of dexamethasone in one sector and injections of physiological saline in the other sector on alternate days. After combinations of dexamethasone (0.25-0.77 mg/kg, i.p.) with the non-preferred sector, rats showed dose-dependent place preference on test day 2; however, when dexamethasone was combined with the initially preferred sector, animals given dexamethasone at a dose of 0.75 mg/kg showed only a slight level of place avoidance. Subthreshold doses of phenamine (0.25 mg/kg) given alone had no effect; however, when given on a background of dexamethasone (0.25 mg/kg), phenamine evoked a place preference response in rats lacking an initial place preference for one sector. Administration of dexamethasone alone (0.25 mg/kg) in these conditions did not induce place preference. Administration of the dopamine D2 receptor antagonist sulpiride (20 mg/kg) 30 min before combinations of dexamethasone (0.25 mg/kg) with the non-preferred sector completely prevented the acquisition of place preference. Administration of the D1 dopamine receptor antagonist SCH23390 (0.05 mg/kg) had no effect on the acquisition of conditioned preference. These results provide evidence for the involvement of D2 receptors in conditioned place preference induced by dexamethasone. PMID- 12762590 TI - Studies of the structure of glutamate receptor ion channels and the mechanisms of their blockade by organic cations. AB - The structural determinants for blockade of the AMPA and NMDA subtypes of glutamate receptors were studied by analysis of structural-functional relationships in a series of mono- and dicationic compounds. The results showed that the hydrophobic and nucleophilic components of the blocker binding sites are located close to each other in the channel of the NMDA receptor, while they are spatially distant in the channel of the AMPA receptor. Molecular mechanical methods were used to construct models of these channels satisfying these topographic criteria and providing adequate descriptions of the binding of the channel blockers. According to the models, binding of blockers to the NMDA channel occurs in the selective filter of the channel (the N/Q/R site). The nucleophilic region of the AMPA channel is formed by the oxygen atoms of glycine residues in position +2 relative to the selective filter. Identification of the major relationships between the molecular structure of the ion channels of these glutamate receptor subtypes and their blockade by organic cations allows the further synthesis of AMPA and NMDA channel blockers with specified levels of activity and selectivity to be directed. PMID- 12762591 TI - Initiation of locomotor activity in spinal cats by epidural stimulation of the spinal cord. AB - Acute and chronic experiments on lower spinal (T10-T12) cats were performed to investigate the effects of epidural stimulation of the dorsal surface of the spinal cord on the initiation of locomotor activity. A zone located at the border between segments L4 and L5 was identified, stimulation of which induces locomotor activity. The parameters of epidural stimulation of the spinal cord effective in activating the stepping movement generator were identified. Epidural stimulation leading to the initiation of movement activity was shown to depend on intracentral and peripheral mechanisms activating the segmental, intersegmental and propriospinal reflex systems of the spinal cord. A leading role was demonstrated for the propriospinal system of the dorsolateral funiculi in activating the generators of stepping movements in epidural stimulation of the spinal cord. PMID- 12762593 TI - Behavioral impairments in acute and chronic manganese poisoning in white rats. AB - Single p.o. doses of manganese chloride (MnCl2 x 4H2O; 50 mg/kg) induced significant and reversible decreases in total activity in white rats, along with worsening of the acquisition of an avoidance reaction in response to unconditioned and conditioned stimuli, increases in the latent period of conditioned reflex activity, and a temporary worsening of the learning process. Chronic manganese poisoning (daily p.o. manganese chloride at 20 or 50 mg/kg for one month) led to significant impairment of learning processes in a multipath maze but had no significant effect on reproduction of previously acquired stereotypical behavior. PMID- 12762594 TI - The effects of stages of the estrous cycle on pain thresholds in female white rats. AB - The effects of stages in the estrous cycle on electrical pain thresholds were studied in white rats in conditions of chronic measurement. On recording day 3, females in the diestrus and estrus stages showed sharp increases in shudder and paw-shuffling thresholds. Females in proestrus and metestrus showed no change in thresholds as compared with those in the first days of recording. Starting from day 7, there were progressive decreases in electrical pain thresholds regardless of the stage of the estrous cycle. Data for all animals showed that regular measurement of pain thresholds induces a smooth two-fold increase in body resistance from day 1 to day 7. This was followed by a sharp drop on day 8, with parallel decreases in body resistance and electrical pain thresholds. Thus, chronic measurement of electrical pain thresholds and body resistance changed in a complex fashion, and threshold values depended on the stage of the estrous cycle. PMID- 12762592 TI - Neurochemical characteristics of the ventromedial hypothalamus in mediating the antiaversive effects of anxiolytics in different models of anxiety. AB - In experiments on rats using an "illuminated area" avoidance test and a "threatening situation" avoidance test, preliminary i.p. administration and subsequent microinjection into the ventromedial hypothalamus of various combinations of monoamines, transmitter amino acids, and their agonists and antagonists demonstrated differences in the functional importance of the neurochemical profile of this limbic formation in mediating anxiety states of different origins. The neurochemical analysis with local intrahypothalamic administration of anxiosedative and anxioselective substances showed that the antiaversive actions of Campirone are obtained only in conditions in which the dominant motivation is fear, while chlordiazepoxide, Phenibut, and Indoter are also active in anxiety induced by negatively stressful zoosocial influences; these actions are mediated respectively by serotoninergic and GABAergic types of synaptic switching in the ventromedial hypothalamus. PMID- 12762595 TI - The ability of new non-competitive glutamate receptor blockers to weaken motor disorders in animals. AB - The ability of mono- and dicationic phenylcyclohexyl derivatives, which are non competitive glutamate antagonists, to prevent convulsions induced in mice by intragastric NMDA or kainate, to weaken catalepsy induced in rats by haloperidol and to exert their own influences of movement activity and behavior in animals was studied. The actions of study compounds were compared with those of the known NMDA antagonists memantine and dizocilpine. NMDA-induced convulsions were effectively prevented by both mono- and dications, while only dications were effective against kainate convulsions. Anticataleptic activity was significantly more marked in monocations, which lacked the ability to block non-NMDA receptors. Side effects on motor coordination were less marked with study compounds than with dizocilpine. Thus, the effects of phenylcyclohexyl derivatives in in vivo experimental models correlate with their anti-NMDA and anti-AMPA activity. They can be regarded as potential agents for treating parkinsonism and other motor disorders. PMID- 12762596 TI - Structural organization of the nervous system of the mantle cavity wall and organs in prosobranch mollusks. AB - A variety of common histological stains was used, along with the Golgi and Colognier silver impregnation methods and electron microscopy, to perform a comparative study of the morphological characteristics of receptor and nerve cells and their interactions in the nervous system of the wall of the mantle cavity and mantle derivatives - gills, siphon, and osphradia - of the marine prosobranch gastropod mollusks Buccinum undatum and Littorina littorea. The results are discussed along with our own previously obtained data on the organization of the osphradial sensory organs and the afferent elements of the mantle cavity wall in other prosobranch mollusks - the terrestrial Pomatia elegans and the freshwater Viviparus contectus and Pomacea paludosa. Using the nervous system of the complex of mantle organs of prosobranch mollusks as examples, the structural features and evolutionary trends of the afferent part of the visceral nervous system of gastropods are discussed. PMID- 12762597 TI - Interaction of histamine and glucocorticoids with neural structures of the respiratory tract. AB - The effects of dexamethasone on the actions of histamine on isolated tissue and large bronchus preparations and the interactions of these substances with intramural neural structures were studied. Low histamine concentrations (10(-12) 10(-8) g/ml) decreased muscle responses induced by stimulation of preganglionic nerve fibers, while high concentrations (10(-7)-10(-4) g/ml) increased these responses. Dexamethasone at concentrations of 10(-7)-10(-6) g/ml decreased muscle responses, while concentrations of 10(-5)-10(-6) g/ml produced biphasic changes in responses. Dexamethasone decreased the effects of histamine at high concentrations. Atropine eliminated the effects of simultaneous application of histamine and dexamethasone on respiratory tract preparations; hexamethonium blocked the effects of substances associated with decreased responses and had virtually no effect on those potentiating responses. Novocaine eliminated the actions of histamine at low and high concentrations and the dilatory effect of dexamethasone. These experimental results led to the conclusion that changes in the responses of muscles from the rat respiratory tract induced by stimulation of preganglionic nerve fibers were modified by low concentrations of histamine and dexamethasone and that these modifications were associated with interactions of these substances with tracheobronchial receptors. PMID- 12762598 TI - Non-synaptic integration of the cell bodies of neurons into the central nervous system of the snail. AB - Single neurons (soma and proximal process) were isolated from the serotonergic (5 HT-ergic) PedA cluster in experiments on the pond snail Lymnaea stagnalis, and changes in the electrical activity of isolated neurons were observed during repeated movement of these cells towards and away from the surface of the CNS. The position of cell bodies of 5-HT-ergic neurons had excitatory effects on the isolated neuron. This effect was maximal (at 10(-8)-10(-7) M 5-HT) when neurons were brought close to the PedA cluster and were further enhanced by addition of the 5-HT precursor 5-hydroxytryptophan at concentrations of (1-2) x 10(-4) M. The results obtained here provide evidence 5-HT-ergic neurons cooperate during 5-HT dependent behaviour, this being based on excitatory interactions at the level of cell bodies. PMID- 12762599 TI - Addressing the caries andd carbohydrate conundrum. PMID- 12762600 TI - Why dentists shun Medicaid: impact on children, especially children with special needs. AB - The Medicaid dental program was established to ensure that needed dental services were obtainable for poor children and adults. General Accounting Office reports and related studies, however, provided a detailed review of the consequences of the economic limitations on the effectiveness of this program. Emphasis in this review was placed on the reluctance of dental practitioners to participate, the resultant limited use of dental services, and the particular problems of youngsters with special health care needs. PMID- 12762601 TI - A comparative study of plaque mutans streptococci levels in children receiving glass ionomer cement and amalgam restorations. AB - PURPOSE: The restorative materials amalgam (Standalloy F) and glass ionomer cements (Chelon Silver) were comparatively investigated to determine the number of mutans streptococci METHODS: Saliva and plaque were collected from patients before and 40 days after the insertion of their restorations. RESULTS: Total bateria and mutans streptococci counts were found to be statistically significantly reduced when compared with the prerestoration counts in the saliva samples (P<0.001). Microbiological analysis of the dental plaque showed that the number of mutans streptococci in the glass ionomer cements was significantly lower than in the amalgam restorations (P<0.001). CONCLUSIONS: This study showed that silver glass ionomer cements inhibited the growth of mutans streptococci. PMID- 12762602 TI - Evaluation of clinical and microbiological features of deep carious lesions in primary molars. AB - PURPOSE: Concern about the survival of microorganisms in deep carious lesions may often lead to unnecessary exposure of the pulp during excavation. This study evaluated the relationship between microbiological and clinical results of deep carious lesions in primary molars. METHODS: Clinical evaluation was performed on 72 deep carious lesions considered to have pulp perforation after traditional excavation. The dentin color and consistency were assessed by means of standardized scales using the technique of Bjorndal et al before restoration. For microbiological examination, dentin samples were taken by a sterile bur and transferred to transport fluid, then plated on tryptic soy agar for growth of total colony forming units. Samples werethen transferred on Rogosa SL agar for growth of oral lactobacilli and on mitis salivarius agar for growth of mutans streptococci. RESULTS: The proportion of mutans streptococci, lactobacilli, and total colony forming units increased when the dentin color and consistency increased. However, the proportion of lactobacilli was smaller than that of mutans streptococci in carious dentin samples. CONCLUSIONS: Contrary to expectations, the findings showed that the number of bacteria in carious dentin of primary molars was not significantly excessive. PMID- 12762603 TI - Effect of APF Minute-Foam on the surface roughness, hardness, and micromorphology of high-viscosity glass ionomers. AB - PURPOSE: The purpose of the study was to evaluate the effect of Oral-B APF Minute Foam on the surface roughness, hardness and morphology of high-viscosity glass ionomer cements (GICs). METHODS: The GICs used were Fuji IX GP and Ketac-Molar. The controls were Vitremer resin-modified GIC and Fuji II conventional GIC. All materials were mixed to a restorative consistency. The encapsulated GICs were mixed with a Rotomix for 10 seconds. For each GIC, 14 specimens (6 mm in diameter and 3 mm thick) were made with a Teflon mold and a fresh mix for each sample. The specimens were allowed to set at room temperature for 15 minutes, and then stored in water at room temperature for 48 hours. Profilometry and microhardness measurements (Knoop) were obtained on the untreated specimens as baseline data. The specimens were then rinsed with water, gently air-dried and 1.23% APF Minute Foam was applied for 1 minute with a brush, rinsed with water, gently air-dried, and hardness and roughness remeasured. In another set, similar measurements were obtained as before but after a 4 minute application of the APF foam. In a different set similar measurements were obtained as before but after a simulated 2-year application of the APF foam. The data were statistically analyzed using an ANOVA. The Student-Newman-Keuls method and Bonferroni tests (P=.05) were used for multiple comparisons of the means. The SEM was used to evaluate the APF effects on the surface micromorphology of the materials. RESULTS: Foam application time had no statistically significant effect on the surface roughness of Ketac-Molar and Vitremer. Fuji IX GP showed that 1 and 4 minute applications had lower values than after 2 years. Fuji II revealed similar roughness for the control at 1 and 4 minutes, but higher values after 2 years. Fuji IX GP proved no significant difference in hardness after the different application times. Ketac-Molar demonstrated less hardness than the control at 1 and 4 minutes but higher after 2 years. Foam application time had no statistically significant effect on the surface hardness of Vitremer. Fuji II showed 1 minute had harder values than 4 minutes and 2 years; the control was harder than 2 years. CONCLUSIONS: The SEM observations revealed that surface micromorphology was not significantly affected by the use of the Oral-B APF Minute-Foam. Treatment with the APF foam tested may be material dependent. It seems to be preferable to apply them for shorter times (1 minute versus 4 minutes) to reduce any adverse effect. PMID- 12762604 TI - Marginal microleakage of two fissure sealants: a comparative study. AB - PURPOSE: An in vitro study was conducted to evaluate enamel marginal microleakage following acid etching of two fissure sealants of established clinical efficacy. The purpose of this study was to assess sealant leakage following etching in combination with the use of latest-generation adhesives. METHODS: Clinicians removed 22 human premolars without caries for orthodontic reasons, stored them in distilled water, and sealed the occlusal surface. They used sealants Concise (3M) and Dyract seal (Dentsply DeTrey). They randomly distributed the premolars into two groups (N = 11) according to sealant used. The materials were manipulated according to the instructions of the manufacturer, and no invasive enamel preparing technique was used (enameloplasty). Staining was carried out with 2% fuchsin. The results were evaluated by the Mann-Whitney nonparametric test for the comparison of independent data samples. RESULTS: Concise sealant was seen to produce significantly greater marginal leakage than Dyract seal (P=.02). CONCLUSIONS: The application of an adhesive layer beneath the sealant afforded less microleakage than when no adhesive was used. Nevertheless despite the possibility of combining adhesives and sealant resins, with good bonding between both components over the entire fissure interface, microleakage was still observed, regardless of the material used. This suggests that presently there is no material is able to secure total sealing of the space between the material used and the dental tissue. PMID- 12762605 TI - Early extraction of the mandibular third molar in case of eruption disturbances of the second molar. AB - PURPOSE: General consensus has been reached on extracting mandibular third molars when they are involved in pathologic processes. However, early extraction is advisable in cases of eruption disturbances of the mandibular second molar, before or during orthodontic treatment. METHODS: Thirty-two patients with early extraction of the mandibular third molar before or during orthodontic treatment were examined. RESULTS: In these cases either the presence of the third molar presented an obstacle in the eruption path of the second, or third molar germ removal was considered beneficial to the course of orthodontic treatment. CONCLUSIONS: Early extraction of the mandibular third molar facilitates the eruption of the second molar, especially in cases where evidence of crowding and lack of space in the posterior mandibular region exist. Orthodontic treatment may, in some cases, aggravate eruption disturbances of the second molar. PMID- 12762606 TI - Association of occlusion with eating efficiency in children with cerebral palsy and moderate eating impairment. AB - PURPOSE: This study focused on the contribution ofthe dental occlusion to chewing efficiency in a group of children with cerebral palsy (CP). METHODS: Twenty children aged 8.3 +/- 0.9 years participated in this study. Chewing efficiency was measured and biting and drinking skills were noted. Occlusion was determined from extra- and intra-oral photographs and from orthodontic plaster models. RESULTS: Significant correlatons were found between chewing efficiency and age (r:0.6, P=.006) and the total number of posterior teeth (r:0.7, P<.001). A marginally significant negative correlation was found between the total number of teeth and chewing time for the viscous texture (r:-0.4, P=.06). CONCLUSIONS: A possible association between chewing efficiency and occlusal wear may exist, but will need further examination. There was also a higher frequency of drooling (23%) in children who had an open bite compared to those with a normal bite. These results suggested that occlusal problems affected the efficiency of ingestion in children with CP. PMID- 12762607 TI - Oral conditions in children with cerebral palsy. AB - PURPOSE: Cerebral palsy (CP) is a major severe childhood disability and its prevalence is increasing, particularly among premature low-weight newborns. The oral conditions observed demonstrate the need for home and professional care for these individuals. This study aimed to investigate dental caries, plaque index, time of eruption, malocclusion, bruxism and motor oral skills in a group of 62 CP children (study group). METHODS: The results were compared to those obtained for a control group with 67 normal children. Nonparametric statistical tests including the Mann-Whitney test, Fisher's Exact test (P), Chi-square test (chi2), and Cochran's Q test were used according to the nature of the variables studied, with the level of significance set at P<.05. RESULTS: The results showed a significantly higher DMFS index for CP children with permanent dentition of both sexes when compared to normal children. A significantly higher plaque index was also observed in the study group. A high tendency to delayed eruption of permanent molars and significantly higher percentages of malocclusion and bruxism were observed in the study group with permanent dentition. Residual food and mouth breathing have been frequently found in CP children. CONCLUSIONS: Earlier preventive measures for CP patients are required because they are a high-risk group for dental caries. PMID- 12762608 TI - Cost-efficiency of a community dental delivery program for migrant children. AB - PURPOSE: The purpose of this study was to describe an economically efficient dental prevention and treatment program aimed at migrant children, using dental school students. This dental treatment program aimed to provide comprehensive dental care to migrant children and to provide a unique learning experience for dental students expanding their knowledge of community delivery systems. METHODS: Twenty-four dental students treated 330 migrant children through a 6-week outreach treatment program sponsored by the University of Michigan School of Dentistry, as a part of the Northwest Michigan Migrant Health Program. RESULTS: The program cost $39,907 and resulted in just over $16,000 of Medicaid reimbursements. Factoring in the Medicaid reimbursements, the program cost $71.58 to treat each child comprehensively. CONCLUSIONS: In light of the reported oral health disparities throughout the United States, this program provides an alternative mechanism for efficient and cost-effective oral care in a group that has traditionally had poor oral health. PMID- 12762609 TI - The relationship of application time to EMLA efficacy. AB - PURPOSE: This study compared the efficacy of 5% Eutectic Mixture of Local Anesthetics (EMLA) cream at application times of 3 minutes, 5 minutes, and 10 minutes to determine the most effective duration of topical anesthetic application on normal mucosa. A special instrument was used to apply pressure on the gingiva to obtain a threshold discomfort level in grams before and after the topical delivery. METHODS: Twenty mL of EMLA cream or a placebo was placed on the maxillary anteriorregion using Beckman paper wicks in the form of discs. The gingiva was wiped dry to get the most effective absorption result. The discs were left on the gingiva for 3, 5, or 10 minutes and off for another 3 minutes. The instrument applied pressure progressively, and the pressure application was stopped when the subjects reported the initial feeling of discomfort. RESULTS: The results indicated that 5% EMLA cream significantly reduced the pain threshold level with 3-, 5-, and 10-minute application times. There was no significant difference among the different durations of 5% EMLA application. CONCLUSIONS: Use of 5% EMLA could be especially beneficial to pediatric patients and young adults who are needle phobic. Development of an EMLA patch with the proper pediatric dosages to prevent possible overdosage and side effects should be the aim of future research. PMID- 12762610 TI - Tooth crown fractures in 3-year-old Andalusian children. AB - PURPOSE: The purpose of this study was to determine the prevalence of tooth crown fractures affecting primary incisors of 3-year-old Andalusian children. METHODS: Clinicians examined 337 three-year-old children (mean age=3.6 years) of Seville, Andalusia, southern Spain. They examined the children for several dental and oral conditions, including tooth trauma. RESULTS: Crown fractures were found in 15% of the examined children. The prevalence of tooth crown fractures was significantly higher in males (18%) than in females (10%) (P=.04; odds ratio:0.5). In both genders, the upper central incisors had the most fractures, 77% in males and 71% in females. The most common crown fracture was found in enamel only (82%), followed by fracture into dentin (12%), and fracture involving the dental pulp (7%). CONCLUSIONS: Dentists have a responsibility to examine primary tooth injuries at the initial presentation of children because a dental injury to a primary tooth may include more severe injuries, such as dento-alveolar fractures. PMID- 12762611 TI - Mesiodens: a clinical and radiographic study. AB - PURPOSE: The term 'mesiodens' refers to a supernumerary tooth present in the midline of the maxilla between the two central incisors. This study seeks to investigate the characteristics of mesiodens among children in Korea. METHODS: The study population consisted of 40 children whose ages ranged from 4 to-26 years. The characteristics of mesiodens were obtained from radiographs and clinical examination results, and were confirmed by surgical intervention. RESULTS: Results showed that males were affected approximately 4 times as frequendy as females. Twenty-five percent of the patients had more than 1 supernumerary tooth. About 66% of the mesiodens were conical in shape and about 52% of the mesiodens were in the upward position. The age and sex distribution, number of mesiodens per patient, shape, direction, size, and effect on permanent maxillary incisors are also presented in this study. CONCLUSIONS: The incidence of mesiodens has been estimated at 0.15% to 1% of the population. It occurs more frequently in boys than in girls, with the ratio being approximately 2:1. In this study, a male:female ratio of 4:1 for the prevalence for mesiodens was found. Ten (25%) out of the 40 patients had 2 mesiodens. PMID- 12762612 TI - Evidence-based restorative dental care for high-risk children. AB - The purpose of this paper was to review the evidence available to the clinician in the area of restorative dental care for high-risk children to determine which approach should be taken in treatment planning for these patients. A literature review was conducted to analyze the aras of population risk, individual risk, treatment outcomes, and decision-making. It also attempted to integrate information from these areas into a plan of action for practitioners trying to understand how these studies should be applied to their practices. PMID- 12762613 TI - A clinical study of the "open sandwich" technique in pediatric dental practice. AB - PURPOSE: This study evaluated the clinical efficacy of the "open sandwich" restoration for pediatric dental practice. METHODS: Three pediatric dentists used a standardized preparation and restorative technique to place the restorations. The prepared tooth was etched with a phosphoric acid semigel and rinsed. A resin modified glass ionomer (Fuji II LCor Photac-Fil) was placed short of the margins and then light cured. The resin modified glass ionomer was covered with an occlusal layer of a microhybrid flowable composite (Aeliteflo or Flow-it). The same preparation for the experimental restorations was used for the control conventional amalgam (Tytin) restorations. The restorations were evaluated at 6 month intervals and ranked with a modified United States Public Heath Survey (USPHS) scale as follows: Alfa: No discernible marginal opening or stain; Beta: Slight opening of margin discernible with dental explorer, but without stain; Charlie: Open margin and stain; Delta: Recurrent caries or restoration failure. Restoration failures were categorized according to etiology, pulpal necrosis, bruxism, marginal leakage, isthmus fracture, or adhesive failure. RESULTS: All recalled experimental restorations, except 8, were rated as either Alpha or Beta. Six failed due to isthmus fracture and 2 due to pulpal necrosis. Fifteen restorations had delaminating of the flowable composite from the resin modified glass ionomer. The use of the "open sandwich" technique compared favorably with a similar study using adhesive amalgam restorations. CONCLUSIONS: The "open sandwich" technique can be successfully used in a pediatric dental practice. PMID- 12762615 TI - Mandibular bilateral fusion in primary dentition: case report. AB - Fusion has been described as a developmental anomaly characterized by the union of 2 adjacent teeth. Bilateral dental fusion in the primary dentition is a rare dental anomaly. The purpose of this report was to present a 9-year-old boy who had bilateral fusion of his mandibular primary lateral incisor and canine teeth. PMID- 12762614 TI - Pulpal therapy for primary teeth: formocresol vs electrosurgery: a clinical study. AB - Post-operative clinical and x-ray findings on 80 primary molars were evaluated. These molars had undergone pulpal therapy by 2 techniques: pulpotomy with formocresol and with electrosurgery. The sample was selected from children between the ages of 4 and 7. A criterion for inclusion was each child must have at least 2 primary molars with pulp exposed by caries. Experimental molars with 1 or more of the following clinical or radiographic signs and symptoms were excluded: mobility, sensitivity to percussion, the presence of fistulae, groove drainage, apical or internal resorption, fetid smell and compromised furcation. Also excluded were patients previously diagnosed with diabetes, cardiopathy and renal alterations. Initially, a dinical and radiographic examination was carried out, followed by another at 1, 3 and 6 months after application of the techniques. No significant differences were found at the end of the period. However, electrosurgery had an advantage: it was faster to apply and dispensed with the use of formocresol. Further research is suggested, using larger samples and shorter evaluation periods. PMID- 12762616 TI - The treatment options of dens invaginatus complications in children: report of 3 cases. AB - PURPOSE: The authors described 3 cases of dens invaginatus accompanied by different periapical complications in children, as well as the therapy methods they used. METHODS: The 3 children were between the ages of 12 and 16 years. The type of dens invaginatus was classified according to Schulze and Brand. RESULTS: All 3 cases had dens invaginatus on their permanent teeth in the maxilla. The complications occurred while the teeth were growing, and they were accompanied by swelling in the region of the dens invaginatus. CONCLUSIONS: The periapical complications required early diagnostic and endodontic treatment to prevent further difficulties at a later stage. PMID- 12762617 TI - Prepubertal periodontitis: a report of 2 cases. AB - PURPOSE: Prepubertal periodontitis is a term suggested for a form of periodontitis that starts soon after the eruption of primary teeth. There is increased awareness that the disease entity known as "periodontitis" is in all probability a family of related but reasonably discrete diseases. The purpose of this report is to describe the clinical, radiographic, and laboratory findings in 2 patients, as well as treatment of prepubertal periodontitis. METHODS: This study presents 2 cases of generalized prepubertal periodontitis: a 5-year-old girl and a 13-year-old girl. RESULTS: Case 1 presented to the clinic because of parental concern for the early exfoliation of her teeth. All of her teeth with Grade III mobility, ie, 8.5, were extracted and the rest were scaled and root planed with subgingival irrigation of 0.2% chlorhexidine. She received a program of 3-month maintenance scaling and 6 months later, there was no further progression of periodontal lesions. Case 2 demonstrated the destruction of tooth roots along with extensive loss of tooth roots and alveolar bone. Since all the permanent teeth were Grade II mobile, the patient was advised to have total extraction with replacement of teeth by complete denture prosthesis at a later date. CONCLUSIONS: In case 2 premature root resorption was occurring concurrently with unexplained extensive alveolar bone loss. In Case 1, the elimination of the pathogenic microflora during primary dentition by early extraction of the teeth involved, plus daily local, mechanical and chemical control of the irritant factors may have prevented subsequent damage to remaining teeth in prepubertal periodontitis. PMID- 12762618 TI - Holistic approach of a child with cystic fibrosis: a case report. AB - The Pediatric Dentistry Clinic staff of the Federal University of Santa Catarina (UFSC) treated a 5-year-old patient who had multiple dental treatment necessities. He also had cystic fibrosis (CF). As CF is an incurable disease that affects the mucous glands, especially the ones related to the lungs and digestive organs, the patient presented both chronic respiratory and intestinal problems. Such problems resulted in a severely compromised nutritional status. Patients who have CF may present problems in oral health because they receive food supplementation that is rich in carbohydrates. Behavioral changes of such patients when in contact with health professionals are also a point to consider because these changes may make the treatment difficult. That is the reason why oral complications imposed by the disease, as well as the psychological aspects related to it, need to be discussed so that treatment is given to the patient aiming at physical and emotional comfort. PMID- 12762619 TI - Adherence to structured physical exercise in overweight and obese subjects: a review of psychological models. AB - Regular physical exercise is a widely accepted means of reducing mortality and improving a number of health outcomes, which is also efficacious in the treatment of obesity. Unfortunately, despite the evidence of favourable short- and long term effects, the results seem to be affected by a lack of adherence, particularly over the long term. It is very important to evaluate and predict good adherence in order to be able to assess the real efficacy of physical activity in the treatment of various diseases. In this paper, we describe most of the psychological models used to evaluate such adherence, and investigate the physiological and psychological bases of physical activity, the most suitable type of exercise for obese patients and some useful strategies for ensuring long term adherence. PMID- 12762620 TI - Assessment of two self-reporting questionnaires for eating disorders in women. AB - OBJECTIVE: The goals of this study were to evaluate how well the Eating Disorder Inventory (EDI) predicted specific eating disorders as identified by the Survey of Eating Disorders (SED), and to identify the most predictive EDI scales. An additional goal was to evaluate the EDI as a screening instrument for eating disorders among a large non-clinical group of women. METHOD: The community-based study sample consisted of 1245 Norwegian women between age 18 and 50 years (mean 23). Subjects filled out the EDI and the SED. RESULTS: The strongest predictors of SED-defined eating disorders were the EDI scales bulimia (BN) and drive of thinness (DT) and body mass index (BMI). When we classified women with an EDI sum score of 40 or higher as having an eating disorder (ED), the sensitivity was 38% and the specificity was 90%, compared to the SED diagnosis of ED. DISCUSSION: Our findings suggest modest agreement between the EDI and the SED, and modest sensitivity and specificity of the EDI. While these instruments, perhaps with some modification, may be adequate for use in selected research applications, they should probably be supplemented with additional information in a clinical or treatment setting. PMID- 12762621 TI - Racial/ethnic differences in weight concerns: protective and risk factors for the development of eating disorders and obesity among adolescent females. AB - OBJECTIVE: This study compared African American and Caucasian adolescent girls on measures of appearance concerns, body-contingent self-esteem, pressures for thinness, and dieting behaviour. Also, correlations among these measures were contrasted across the two ethnic groups. METHOD: African American (n = 32) and Caucasian (n = 29) adolescent girls completed a series of self-report measures concerning dieting behaviour, perceived pressures for thinness, self-esteem, and perceptions of physical appearance. RESULTS: Across all measures, Caucasian girls endorsed more disturbed eating behaviors and beliefs about body shape and weight. Dieting among Caucasian girls was correlated with preoccupation with body shape. Among African American girls dieting behaviour was unrelated to concerns about body shape. DISCUSSION: These findings support the hypothesis that in comparison to Caucasian girls, African American girls may be at lessened risk for developing eating disorders but may be at greater risk for developing obesity. This conclusion is based upon the findings of higher levels of eating disorder symptoms among Caucasian girls and the relative absence of significant correlations between dieting and body shape and appearance concerns among African American girls. PMID- 12762622 TI - Expressed emotion, family environment, and parental bonding in bulimia nervosa: a 6-year investigation. AB - As part of a prospective, long-term treatment study, 30 in-patients with bulimia nervosa (BN) were divided into groups with high and low expressed emotion (EE) family backgrounds according to the Camberwell Family Interview, and followed for a period of six years. The high EE group initially showed significantly more psychopathology than the low EE group. No group x time interactions were found, but the high EE group showed a worse outcome on the "conflict" and "organisation" subscales of the Family Environment Scale. They also showed significantly more eating disorder pathology according to the Eating Disorder Inventory (EDI) and the Structured Interview for anorexia nervosa (AN) and BN before treatment at discharge, after two years and, to some degree, even after six years. Depth of depression (Beck Depression Inventory) was significantly higher in the high EE group at admission (moderate depression), discharge and after the 6-year follow up (still slight depression). The Parental Bonding Instrument (PBI) showed no differences between the high EE and low EE groups, but the individuals with "affectionless control" according to the PBI had more negative scores on three of the subscales of the Family Environment Scale (FES). In brief, the high EE individuals with BN were initially sicker and did not fully catch up over time in comparison with the symptomatic recovery of the low EE individuals. These data suggest that EE status upon admission to in-patient treatment is a relevant predictor of the severity and course of BN and depressive symptoms. PMID- 12762623 TI - Effects of different training protocols on exercise performance during a short term body weight reduction programme in severely obese patients. AB - The effects on aerobic, anaerobic and strength performance of aerobic (A) or A and strength (AS) training, integrated with a 3-week body weight reduction programme involving an energy-restricted diet, nutritional education and psychological counselling, were evaluated in 52 grade II or III obese individuals (A: n = 26; AS: n = 26). After 3 weeks, both training programmes led to a similar and significant reduction (p < 0.001) in body mass (A: 4.8%; AS: 4.4%) and an increase (p < 0.05 - p < 0.01) in maximal oxygen consumption (A: 16.8%; AS: 10.9%). A significant (p < 0.05 - p < 0.001) increase was found in absolute lower limb anaerobic power output evaluated by means of a jumping test (A: 13.7%; AS: 18.1%) and stair climbing test (A: 9.7%; AS: 4.3%), without any significant difference between A and AS. A comparable and significant (p < 0.01) increase was also found in average horizontal velocity during a short sprint running test (A: 8.1%; AS: 7.1%), with a shorter time of foot-ground contact (A: 4.9%; AS: 6.6.%) and a higher step frequency (A: 4.0%; AS: 10.4%). The maximum strength increase after the body weight reduction programme determined by one maximal repetition test of lower and upper limb muscle groups was significantly greater (p < 0.05 - p < 0.001) in the AS group, ranging from 11.4% to 25.4% (A) and from 26.7% to 41.8% (AS). These results indicate that integrating a body weight reduction programme involving diet nutritional education and psychological counselling with A or AS exercise has similar positive effects in lowering body mass and improving A and anaerobic performance. However, the addition of strength training to A conditioning increases maximum strength. PMID- 12762624 TI - Acculturation and eating disorder symptomatology in Black men and women. AB - OBJECTIVE: We assessed the prevalence of eating disorder symptoms in Black South African men and also explored their possible links with acculturation. METHOD: The Eating Disorders Inventory (EDI) and the South African Acculturation Scale (SAAS) were administered to a sample of undergraduate students consisting of 50 Black women, 50 Black men and 50 White men. RESULTS: The Black men had significantly higher scores than the White men on the psychological subscales of the EDI and higher than cut-off scores on perfectionism, interpersonal distrust and maturity fears subscales. There were marginal racial differences in the average acculturation scores, whereas gender appeared to have a variable effect on acculturation attitudes. Specific acculturation strategies were found to be differentially associated with the EDI subscales. The association between acculturation and eating disorder symptoms in Black men was significantly higher than that in White men. DISCUSSION: Our results indicate that there is a higher prevalence of eating disorder pathology in Black men than in White men. This lends support to the possible association between acculturation and eating disorders, and challenges the assumption that non-Western individuals--and Black men in particular--are protected from body image disturbances and disordered eating patterns. PMID- 12762625 TI - Urinary peptide levels in women with eating disorders. A pilot study. AB - The aim of this study was to evaluate whether the urinary excretion of low molecular weight peptides is increased in women with a history of anorexia nervosa/self starvation. The study group consisted of 12 women aged 20-38 years who were treated in a specialised day care unit for eating disorders in Stockholm between January and December 1998; the controls were eight women with primary bulimia treated in the same unit (A) and ten healthy women without any eating disorder (B). The chromatographically measured urinary peptide levels in the study group were significantly higher than those in control group A (and B when one highly influential individual with very low peptide excretion in the study group was excluded from the analyses). These findings offer some support to the speculative hypothesis that eating disorder symptoms may be linked to increased levels of neuroactive peptides, although it is necessary to define the peptides further before any definite conclusion can be drawn. Furthermore, the study group was characterised by many interpersonal differences in eating behaviour that could explain the increased urinary peptide levels. PMID- 12762626 TI - Psychodynamics of onychophagists. AB - The authors offer three interpretations of onychophagia, a widespread but largely unstudied phenomenon. The first is phenomenological, and characterises onychophagia as a non-pathological behavioural symptom present in many pathologies and a distinct syndrome that is sometimes quite severe. The second is clinical, and indicates the convergent and divergent connections between onychophagia and the most frequent eating disorders. The third is psychodynamic, and suggests an interpretative hypothesis of onychophagia itself and the frequent sensation of trouble and refusal that it often provokes even in doctors and psychotherapists. PMID- 12762627 TI - Silent pericardial effusion in a sample of anorexic patients. AB - Mortality in anorexic patients is mainly due to suicide or cardiac failure. The aim of this study was to investigate structural and functional cardiovascular alterations further by means of echocardiography in a sample of 15 medication free patients with DSM-IV anorexia nervosa (AN) (BMI < 17.5 kg/m2) and without any known cardiovascular disease and/or a family history of deafness or sudden death, and correlate the findings with clinical variables. The controls consisted of a sample of 10 constitutionally thin women (BMI < 19 kg/m2), of comparable age, height and degree of physical activity. All of the subjects underwent Doppler echocardiography (ECHO), and the patients were also administered the Diagnostic Schedule for Eating Disorders (DSED) in order to assess the features and course of the eating disorder. ECHO revealed silent pericardial effusion in 71.4% of the patients vs. 10% of the controls (p < 0.05); among the patients, the separation of pericardial leaflets was more frequent in those with a shorter duration of illness (p < 0.05). Mitral valve motion abnormalities were more frequent among the patients than the controls (69.2% vs. 10%, p < 0.005), and the left ventricular mass/body surface area was lower (54.8% vs. 59%, p < 0.001). Isovolumetric relaxation time was longer in the patients (98.4 vs. 65 msec, p < 0.01), but there were no significant differences in left ventricular ejection fraction (53.8% vs. 59%) or early diastolic deceleration time (146 vs. 155 msec). The results of this study support the association between AN and demonstrable anatomic and functional cardiac abnormalities, such as a reduced ventricular mass and mitral valve abnormalities. The ECHO findings provide evidence for clinically silent pericardial effusion in AN, which may be an early sign of cardiovascular involvement. PMID- 12762628 TI - Shame, depressive symptoms and eating, weight and shape concerns in a non clinical sample. AB - Shame has been shown to be related both to symptoms of depression and eating pathology. However, the independence of this relationship has not yet been established. The purpose of the present study was to determine whether the relationship between shame and eating disorder symptoms was independent of the relationships of these variables with depression. Seventy non-clinical female participants completed measures of eating disorder-related concerns using the Eating Disorder Examination-Questionnaire version (EDE-Q), depressive symptoms using the Beck Depression Inventory (BDI-II) and two measures of shame, the Other As Shamer Scale (QAS) and the Test of Self-Conscious Affect (TOSCA). Despite a strong association between BDI-II and EDE-Q scores and a moderate relationship between the shame measures, the two measures of shame showed some specificity in their relationships with symptom measures. The OAS was independently related to levels of BDI-II scores while the TOSCA was independently related to scores on the EDE-Q. There are a number of differences between the two measures of shame used in this study. The fact that each was differentially related to eating concerns and depressive symptoms may give clues as to which aspects of shame are important in each of the two types of pathology. PMID- 12762629 TI - Anorexia nervosa: the cost of long-term disability. AB - This study was performed to estimate the cost of long-term disability in people who have anorexia nervosa (AN) that live in the province of British Columbia (BC), Canada. Canada provides universal socialized health and welfare services, and each of the 10 provinces is responsible for its own funding. As the provincial government of BC does not categorize its disability payments by the cause of the disability, a survey was used to determine the rate of disability from AN. A sensitivity analysis was performed to assess the influence of variations on the yearly cost of disability in BC: the number of patients with AN was varied between 1.0 and 2.0% of the female and 0.05 and 0.1% of the male population; the percentage of patients with AN receiving disability payments was determined by the survey to be 35%; the cost of these payments was varied between the lowest and highest benefits a single person can receive from the BC provincial government; and finally, to allow for possible sampling bias and a possible lower prevalence of AN, the lower limit of the sensitivity analysis was derived by dividing the lowest estimate above by seven. The sensitivity analysis revealed that the total estimated cost of long-term disability in BC could be as low as $2.5 million (Canadian) or as high as $101.7 million per year, which is a cost of up to 30 times the total yearly cost of all tertiary care services for the treatment of eating disorders in BC. In view of this finding, an increase in funding is warranted for primary, secondary and tertiary prevention programs for AN in BC. PMID- 12762630 TI - Satiety after vertical banded gastroplasty. AB - Early satiety following gastroplasty is due to the new anatomic conditions created by the operation and refers to a distressing or painful epigastric sensation after food consumption. Early satiation may act as an aversive stimulus, shaping eating habits and behaviour in such a way as to promote satisfactory weight loss and maintenance. This hypothesis was tested in patients who had undergone vertical banded gastroplasty (VBG). The duration of the sensation of early satiety was associated with the scores of questionnaires assessing eating behaviour, but seemed to be completely unrelated to the radiologically measured proximal pouch volume, energy intake and weight loss data. These findings suggest that cognitive factors play a substantial role in determining food intake and therefore in achieving weight goals following gastric restriction. PMID- 12762631 TI - Family therapy for identical twins with anorexia nervosa. AB - The rarity of identical twins with anorexia nervosa (AN) makes formulating an effective treatment approach for this subset of individuals quite difficult Clinicians may be applying treatment strategies that have been designed for individuals with an eating disorder. However, these approaches might have to be modified for the treatment of identical twins. The few available case studies have not discussed the challenges a clinician faces when treating identical twins with AN. In this report we describe the presentation of the case of adolescent monozygotic twins who were simultaneously concordant for AN. We discuss their unique presentation and our approach to their treatment. PMID- 12762632 TI - Gain of function TSH receptor mutations and iodine deficiency: implications in iodine prophylaxis. AB - Iodine deficiency is widely known to be the main cause of nodular goiter (NG). In iodine deficient areas subclinical and overt hyperthyroidism is the major cause of morbidity and it is mainly due to toxic NG rather than Graves' disease. Toxic NG, including toxic multinodular goiter and toxic thyroid adenoma is usually encountered in subjects with long-standing NG, in whom thyrotoxicosis is usually preceded by a long phase of euthyroidism and then subclinical hyperthyroidsm (abnormally low TSH with normal circulating thyroid hormones). Epidemiological studies indicate that, compared to Graves' disease, the incidence and prevalence of non-autoimmune hyperthyroidism due to toxic adenoma and toxic multinodular goiter differ in different regions of the world, being much more frequent in areas of iodine deficiency. Recently, mutations of the TSH receptor (TSHr) gene causing permanent activation of the thyroid follicular cell adenylate-cyclase, have been shown to be the most probable cause of the hyperfunction and growth of toxic adenoma. In this review we will focus our attention on the role of external factors (i.e. iodine deficiency) with respect to individual factors (i.e. genetic mutations) in the pathogenesis of toxic NG. PMID- 12762633 TI - The impact of iodine prophylaxis on thyroid 131-iodine uptake in the region of Krakow, Poland. AB - Iodine prophylaxis was introduced in Poland in 1935. It was interrupted twice, the first time between 1939 and 1947 (due to the Second World War and its aftermath), and then between 1980 and 1986, due to the economical crisis in Poland at that time. A voluntary model of iodine prophylaxis (20 +/- 10 mg/kg of household salt) introduced in Poland in 1986, was followed by a mandatory model, implemented at the beginning of 1997, with 30 +/- 10 mg/kg of household salt. In the early sixties, in our Department of Endocrinology 24-hour iodine uptake test was introduced as a routine procedure for evaluating the thyroid gland. The reference value of this test increased in 1986 after discontinuation of iodine prophylaxis, and decreased in 1998, two years after re-implementation of the mandatory model of iodine prophylaxis. In 167 patients (147 with endemic goiter and 20 with no thyroid disturbances) examined between 1998 and 2000 the mean value of 24-hour iodine uptake was 27.3 +/- 10.4%, as compared to the mean value of 45,7 +/- 6.6 % in 1986. In patients with thyrotoxicosis the mean value of iodine uptake was 41.9% +/- 16.2 in 1999 (no.=614), 42.4% +/- 16.9 in 2000 (no.=644) and 37,7% +/- 17.2 at the beginning of 2001 (no.=328). The mean value of iodine uptake in patients with thyrotoxicosis before implementation of iodine prophylaxis was over 60%. The 24-hour thyroid radioiodine uptake test proved to be a useful indicator of changes of iodine intake in a iodine-deficient population. The data presented in this study confirm the importance and efficacy of the iodine prophylaxis introduced in Poland. PMID- 12762634 TI - Prevalence of goiter in schoolchildren--a study on the influence of adequate iodine prophylaxis in Poland. AB - Poland has been known as an area with iodine deficiency. Surveys carried out in 1992/1993 and 1994 revealed that the voluntary model of iodine prophylaxis introduced in 1986 was ineffective. In 1997 a new model of iodine prophylaxis based on obligatory household salt iodization has been implemented. In order to assess its effectiveness new studies were undertaken in 1999-2001. The study involved 1471 school-children aged 6-15 years from 12 sites. In every subject thyroid volume by means of ultrasound and urinary iodine concentration were assessed. The results were compared with data obtained from the same schools in the 1992/1993 survey. Between 1992/93 and 1999/2001 goiter prevalence decreased from 14.5% to 5.2% (p<0.05) and median urinary iodine concentration increased from 56 microg/l to 103 microg/l (p<0.05). A decrease in goiter prevalence was observed in 6 sites with moderate goiter endemia, whereas the changes in goiter prevalence were statistically insignificant in other 6 sites. Three sites were characterized by goiter prevalence close to 5% before and after implementing the obligatory model of iodine prophylaxis. Goiter prevalence in the remaining three sites remained within the same limit of 7-10%, in spite of observed ioduria increase. The Authors conclude that the Polish model of obligatory iodine prophylaxis ensures efficient iodine supplementation and this is confirmed by a significant increment in ioduria. The effect of this model on thyroid volume is evident in moderate goiter endemia areas. Slight changes in goiter prevalence in mild goiter endemia regions need further monitoring and considering other factors affecting thyroid volume. PMID- 12762635 TI - Iodine deficiency disorders in the Saratov province in Russia. AB - OBJECTIVE: The work deals with epidemiology of iodine deficiency (ID) in the Saratov province in Russia, with clinical data and thyroid autoimmunity disorders in schoolchildren living in a moderate ID area. STUDY DESIGN: The Saratov province is situated in the Volga Region. The epidemiological study was made according to WHO/UNICEF/ICCIDD recommendations (1994). METHODS: 2040 children aged 6-12 in 21 schools from 14 districts of the province were examined (palpation and ultrasound). Ioduria was measured in 654 children. The results of screening for congenital hypothyroidism in 95657 neonates were analysed. The clinical study included hormonal (tT3, tT4, TSH) and immunological (Abs TPO, Abs Tg) assays in 246 children. RESULTS: In the Saratov province of Russia moderate ID has been proved (ioduria median 38.2 microg/l, endemic goiter incidence in children 35.8% by palpation and 25.8% by ultrasound, hyperTSHemia incidence in neonates' blood 37.4%). Ioduria median correlates with the data of other ID epidemiological criteria in different ways, it being most high with hyperTSHemia. Thyroid nodules were detected in 2.0% goitrous children. Hypothyroidism was found in 31.1% of the goitrous children. Thyroid antibodies were determined in 10.8% of the children. The incidence of antibodies increases with age and goiter size. High prevalence of associated chronic pathology in goitrous children was observed. CONCLUSION: According to WHO/UNICEF/ICCIDD criteria, moderate ID in Saratov province has been found. Iodine deficiency disorders in children area cluster of pathological states, endemic goiter being one of the clinical symptoms. High prevalence of goiter, hypothyroidism, associated chronic pathology in children have been detected. PMID- 12762636 TI - Bioavailability of oral vs intramuscular iodinated oil (Lipiodol UF) in healthy subjects. AB - BACKGROUND: In order to fight against iodine deficiency, the essential cause of endemic goiter and cretinism, several health organizations promoted campaigns of iodinated oil (Lipiodol UF) administration using iodinated oil administered intramuscularly. However, it seems preferable to administer iodinated oil orally, as this is more appropriate and since the efficacy of this route has been demonstrated as well as for intramuscular route by controlled clinical trials. OBJECTIVE: To assess the bioavailability of iodinated oil (Lipiodol UF) administered via two different administration routes and the safety profile of this agent. DESIGN: A randomized bioavailability study was performed comparing a single oral dose of 3 capsules (570 mg of iodine) vs a single intramuscular injection of 1 ml of Lipiodol UF (480 mg of iodine) in 36 healthy subjects followed for 9 months. RESULTS: The results show that, at these dosages, the 24 h urinary iodine values are above baseline for both oral and intramuscular administrations (im: >12 months/oral: 6 months) for prolonged period of time. In terms of safety, Lipiodol, administered by im injection or orally, did not induce any undesirable effects or any alteration of thyroid function tests in this study. CONCLUSIONS: In conclusion, this study shows that im or oral administration of a single dose of Lipiodol provides a significant and prolonged iodine supplement. The results obtained confirm the possibility of protection of exposed populations after annual administration of an appropriate single oral dose, without inducing any clinical or laboratory adverse effects. The product, by either route of administration, has a prolonged efficacy in iodine-deficient subjects (im: 2-3 years/oral: 1 year). PMID- 12762637 TI - Neonatal hypothyroid screening in monitoring of iodine deficiency and iodine supplementation in Poland. AB - Neonatal hypothyroid screening in Poland is standardised and all newborns screening data are registered in central data base in the National Institute of Mother and Child. About 400,000 newborns are screened per year for hypothyroidism (TSH) and phenylketonuria (PKU). Unfortunately, obstetric clinics still use antiseptics that contain iodine. According to our data 71% of clinics used iodine in 1998 (58% iodine tincture and 13% povidone iodine) and 58.2% (35.4 iodine tincture and 13% povidone iodine) in the year 2000. Presence of iodine resulted in over 3 times increase of a percentage of TSH levels over cut off, increasing the number of false positives in the hypothyroid screening. Analysis of TSH distribution for iodine containing and iodine free hospitals gave totally different estimation of iodine deficiency according to WHO criteria. In the group of iodine free hospitals 24 regions were classified as not deficient, 9 regions were borderline with a fraction of TSH levels over 5 mlU/l of 3-5%. 10 regions could not be analysed because all hospitals declared use of iodine. In the second group all regions were iodine deficient. TSH distribution since 1994 shows significant decrease of percentage of TSH levels over cut off from 2.23% in 1994 to 0.16 in 1997 and to 0.12 in 2000. These changes are most probably connected with successive introduction of iodine supplementation which became obligatory in 1997 and suggest that iodine supplementation covers iodine requirements during pregnancy. CONCLUSIONS: Iodine deficiency and iodine supplementation in Poland can be studied using TSH blood spot newborn screening results in correlation with data on interfering factors and in reference to modified criteria for the analytical test and the population. To reduce false positive rate in neonatal hypothyroid screening iodine containing antiseptics, particularly iodine tincture, should be withdrawn from all obstetrics clinics in Poland. PMID- 12762638 TI - Iodine deficiency disorders incidence in neonates based on the experience with mass screening for congenital hypothyroidism in southeast Poland in the years 1985-2000. AB - Thyroid hormones are very important for the child mental and physical development. In southeast Poland screening for congenital hypothyroidism detection was established in 1985; this region was defined as an iodine deficient area. The results provided the first signal pointing to insufficient iodine prophylaxis in the region. In the years 1985-2000, a total of 634,179 newborns were tested. TSH levels in blood spots on filter paper were determined using three consecutive methods: RIA, fluoroimmunometry (FIA) and luminoimmunemetry (LIA). In the first five years of the program, the incidence of permanent hypothyroidism was 1:4,000, while the incidence of iodine deficiency disorders (IDD) was 1:272. These findings led to the recognition of the fact that iodine prophylaxis had been discontinued in 1981 and reintroduced in 1992 and from that time on the incidence of IDD in neonates has been evidently lower. In the period between 1991 and 2000, we observed a 0.14% incidence rate of IDD, while until 1991 it had been 0.5%. In the years 1998-2000, the respective value was 1:5,420. The incidence of transient hypothyroidism also dropped from 1:3,920 to 1:48,474. Therefore we conclude that, based on screening studies carried out since 1985, the analysis of TSH levels may be regarded as a tool for iodine deficiency monitoring in neonates. Between 1985-2000, a drop in the incidence of IDD in newborns is clearly seen. Moreover, investigations carried out in particular years show that even low-grade iodine supplementation leads to TSH level decrease in newborns. PMID- 12762639 TI - Changes of urinary iodine excretion of newborns over a period of twenty years. AB - OBJECTIVE: The use of iodized salt is not mandatory in Hungary and supply is irregular since decades. The effect of this anomaly was studied on the neonatal urinary iodine excretion over a period of twenty years. DESIGN: Urinary iodine content of 286 newborns' samples was tested in different areas of Hungary in 1980, 1991, and 1999. METHODS: Colorimetric ceric ion-arsenious acid method based on Sandell-Kolthoff reaction was used for determination of urinary iodine content. Fitfy-one samples were excluded from evaluation because of suspicion of iodine contamination. RESULTS: Decreased iodine excretion was demonstrated in different areas and at various times over this period of twenty years in accordance with the results of other (schoolchildren, pregnants) surveys in Hungary. CONCLUSIONS: Iodine deficiency of newborns in these areas changed from year to year, documenting a fluctuating iodine intake. Universal salt iodization should be introduced throughout the country instead of the present "silent iodine prophylaxis" to prevent iodine deficiency, especially in the perinatal period. PMID- 12762640 TI - Improvement of iodine deficiency after iodine supplementation in schoolchildren of Azerbaijan was accompanied by hypo and hyperthyrotropinemia and increased title of thyroid autoantibodies. AB - OBJECTIVE: In the mountainous areas of Azerbaijan the schoolchildren suffer from severe Iodine Deficiency (ID) with median Urinary iodine excretion (UIE) 36 mcg/l and prevalence of goiter 99% (estimated by US). In a population of 293,000 schoolchildren aged 8-14 y.o. we administered capsules containing 190 mg of iodized oil (Lipiodol-Guerbet, Cedex, France) twice yearly in 6 months apart (total 380 mg). The aim of the present study was to evaluate the efficacy, the benefits, as well as the possible side-effects in a follow-up period of 6 and 12 months after the initial administration of iodized oil. METHODS: Six and 12 months after the initial administration of iodide, two representative samples of 391 and 326 children respectively were examined. The evaluation included: estimation of goiter by US, determination of UIE and serum measurements of T3, T4, TSH, Tg, autoantibodies against thyroid peroxidase (anti-TPO) and thyroglobulin (anti-Tg). RESULTS: There was an improvement in median UIE which increased from 36 mcg/l to 68 and 81 mcg/l after 6 and 12 months of treatment respectively. The prevalence of goiter decreased from 99% to 54% and 26% respectively. Tg was decreased at 6 and 12 months from the first administration, whereas TSH remained unchanged at 6 months and decreased at 12 months when compared to the latter value. Hypothyroidism was detected in 7% of children after iodide administration both at 6 and 12 months, but overt hypothyroidism was observed only in 0.5% at 12 months. Subclinical hyperthyroidism was detected in 2% and 6% after iodide administration both at 6 and 12 months. There was a significant increase in the title of thyroid auto antibodies in 6 months which was retained and increased in 12 months. There was no relation between the appearance of thyroid dysfunction and the positive thyroid auto antibodies. CONCLUSION: The dose of 190 mg iodide administered twice yearly, improved iodine deficiency and endemic goiter in schoolchildren. The increase of UIE resulted from iodide administration, was accompanied by an increased title of thyroid auto antibodies and an increased prevalence of hyper- and hypothyrotropinemia apparently of no autoimmune etiology. PMID- 12762641 TI - Iodine excess and thyroid autoimmunity. AB - Epidemiological studies, as well as animal models, indicate that iodine might be an immunogenic agent for the thyroid gland, at least in subjects predisposed to thyroid autoimmunity. This review presents data, either epidemiological or experimental, obtained in different conditions: constant and stable iodine status, either deficient, sufficient or excessive; long-term iodine prophylaxis; temporary supplementation with iodide (6-12 months) or iodised oil. Moreover, we also discuss data obtained in the general population, among subjects with euthyroid goiter, or autoimmune goiter, or even in women prone to post-partum thyroid diseases. It is concluded that the significant increase in the prevalence of autoimmune thyroid diseases in populations living in iodine sufficient areas should not prevent the implementation of the iodine prophylaxis. PMID- 12762643 TI - Incidence of thyroid cancer in the selected areas of iodine deficiency in Poland. AB - The aim of the study was to evaluate the incidence rate (IR), trend and histotype of the differentiated thyroid cancer in the selected areas with varying iodine deficiency. The study was carried out in three areas: Krakow, (Carpathian endemic goiter area with 1.99 million mixed rural and urban population), Gliwice (Upper Silesia--moderate iodine deficiency area mostly industrial with 4.89 million inhabitants) and Olsztyn (slight iodine deficiency area, mainly rural with 0.77 million inhabitants). Between 1990 and 2001, in the study area 2691 newly diagnosed cases of malignant neoplasms of the thyroid gland were registered. In over 80% of patients it was differentiated thyroid cancer: mainly in women over 40 years, with F/M ratio 5.8. The highest percentage of papillary cancer 72.9% was observed in Olsztyn and lowest--50.0%--in Krakow and Nowy Sacz districts. In this period of time incidence rate of differentiated thyroid cancer in women increased in Krakow, Gliwice, and Olsztyn from 1.51 to 9.34 in 1998 1.27 to 5.74 in 1999 and from 2.52 to 11.35 in 2001 respectively. In the youngest (0-20 years) age group no significant increase of IR was observed. Between 1998 and 2001 the dynamics of increase of the thyroid cancer incidence markedly diminished. In conclusion it was hypothesised that an increase in IR of differentiated thyroid cancer in the study area was caused mainly by the suspension of iodine prophylaxis in 1980 and was diminished by the introduction of an obligatory model of iodine prophylaxis in 1996/1997. It was modified in terms of histotype and dynamics of increase by exposure to ionizing radiation. A very specific group at risk on the population level were women aged 20-40 years in the reproductive age exposed to iodine deficiency after suspension of iodine prophylaxis in 1980 and to radiation after the Chernobyl accident in 1986. PMID- 12762642 TI - Iodine-induced hyperthyroidism--an epidemiological survey several years after institution of iodine prophylaxis in Poland. AB - In 1997, the obligatory model of iodine prophylaxis was introduced in Poland in order to correct the existing status of mild and/or moderate iodine deficiency. In order to monitor possible side-effects of increased iodine supply, studies on iodine-induced hyperthyroidism were initiated by establishing several regional registers of hyperthyroidism. In the present paper, the results of a two-year monitoring (2000-2001) have been summarized. There are no epidemiological data on hyperthyroidism prior to starting the iodine prophylaxis, but the obtained current data are comparable to observations in other countries, made after iodine supplementation. The incidence of iodine-induced hyperthyroidism did not exceed the acceptable level, thus confirming--together with previous observations on the effectiveness of iodine prophylaxis--the adequacy of applied dose of KI (30 microg/kg NaCl), used for salt iodization in Poland. PMID- 12762644 TI - Influence of iodine deficiency and iodine prophylaxis on thyroid cancer histotypes and incidence in endemic goiter area. AB - The aim of the study was to evaluate the correlation between thyroid cancer histotype and incidence rate (IR) and iodine nutrition level in two endemic goiter areas: the districts of Krakow and Nowy Sacz. The suspension of iodine prophylaxis in Poland in 1980 resulted in increased goiter prevalence in schoolchildren and adults and elevated TSH levels in newborns in the early 1990s. Since 1992 a rise in thyroid cancer IR was observed. Thyroid cancer IR in the Krakow population was 2.22 in 1986; 3.62 in 1995 and 6.02 in 2001; in Nowy Sacz: 1.52; 2.59 and 3.88 respectively. In 1986 papillary/follicular cancer ratio in both areas was about 1.0--the value typical of iodine deficient areas. After restoring the obligatory iodine prophylaxis in 1997, a significant decrease in elevated TSH concentration in newborns and urinary iodine concentration increase in schoolchildren were observed. A relative rise in the incidence of papillary thyroid cancer and decrease in follicular cancer, resulting in rise in papillary/follicular thyroid cancer ratio up to 5.9 in 2001 was also observed. Since 1999 no further thyroid cancer IR increase was noted. In conclusion, a significant increase in differentiated thyroid cancer IR was observed in association with the iodine prophylaxis suspension. Changes in thyroid cancer histotypes in 1986-2001 and a significant decrease in incremental rate of differentiated thyroid cancer probably reflect the influence of effective iodine prophylaxis. The significant difference between IR of thyroid cancer incidence in the districts of Krakow and Nowy Sacz may be related to differences in the exposure to radiation after the Chernobyl accident. PMID- 12762645 TI - Organophosphorous pesticides in breast cancer progression. AB - Environmental substances may be involved in the etiology of breast cancers. Many studies have found an association between cancer in humans and exposure to agricultural pesticides. Organophosphorous pesticides have been used to control mosquito plagues. Parathion and malathion, organophosphorous pesticides are cholinesterase inhibitors responsible for the hydrolysis of body choline esters, including acetylcholine at cholinergic synapses. Their primary target of action in insects is the nervous system whereby they inhibit the enzyme acetylcholinesterase at synaptic junction. Atropine is a parasympatholytic alkaloid used as an antidote to acetylcholinesterase inhibitors. We have established an experimental breast cancer model, where epithelial cells in the rat mammary gland underwent a stepwise transformation into malignant cells by exposure to pesticides (Cabello et al, 2001). The aim of this work was to examine whether pesticides were able to induce progression of malignant transformation of a human breast epithelial cell line, MCF7. These results showed that parathion and malathion increased PCNA and induced mutant p53 protein expression of MCF7 cells in comparison to controls and atropine inhibited such action. These results indicated that organophosphorous pesticides can induce more changes in this malignant breast cell line, inducing another step in the progression of the transformation process and atropine on the other hand inhibited the effect of such substances. PMID- 12762646 TI - Collagen fibers in human prostatic lesions: histochemistry and anisotropies. AB - The present study focuses on establishing patterns of collagen fibers distribution in prostatic nodular hyperplasia and adenocarcinomas, in comparison with the normal tissue. Sections of prostatic transurethral resection were subjected to Gomori's method for collagen fibers and reticulin and analyzed under ordinary and polarized light microscopy. Controls and hyperplastic regions present collagen fibers with variable thickness that run in different directions, establishing a tridimensional network. These fibers exhibit birefringence and dichroism thus demonstrating their fibrillar integrity. On the other hand, increased variability in collagen fiber distribution and anisotropical properties occur in adenocarcinomas evaluated in accordance withthe Gleason's score. In some of their areas, a well-defined collagen network delimitates the base of transformed epithelial cells whereas in other areas the collagen fibers are disorganized and do not establish a boundary between the epithelial structures and the stroma. In these areas, collagen is found in the stroma. It was also observed that adenocarcinoma tumor cells rest on a scaffold of thin and dendritic collagen fibers. Collagen fibers of the prostatic stroma of the adenocarcinomas may show a modification in arrangement and fibrillar compactness. In prostatic nodular hyperplasia, there is no change in collagen molecular integrity, since collagen affinity for silver and collagen birefringence are similar to controls. In adenocarcinoma with high dedifferentiation degree, thin and branched strongly argyrophilic and birefringent collagen fibers are detected in regions of cell proliferation. In the adjacent stroma, hyaline plaques are indicative of matrix degradation or remodellation. PMID- 12762647 TI - A case of splenic lymphoma with marked diffuse nodular fibrosis and calcification, complicated with severe autoimmune hemolytic anemia. AB - A splenic lymphoma, possibly of a splenic marginal zone lymphoma, marked by small nodular splenic calcified fibrosis and complicated by intractable autoimmune hemolytic anemia, was studied by immunohistochemical, molecular genetic, and ultrastructural analyses. The patient was a 57-year-old Japanese man who had moderate splenomegaly, and who had undergone splenectomy for improvement of severe autoimmune hemolytic anemia and to rule out malignancy in the spleen. In the resected spleen, proliferative atypical lymphoid cells were observed both in the red and white pulp with diminished germinal centers and irregularly widened marginal zones with peculiar dimorphic pattern. Ultrastructural study revealed no hairy cells or villous lymphocytes. Diffuse nodular hyalinous fibrosis surrounding the small arterioles in the white pulp overlapped with frequent calcification was a unique histologic feature in this case. Degenerative connective tissue, extracellular matrix, or collagen fibers surrounding the arterial sheath in the white pulp caused by some immunological abnormalities associated with this splenic lymphoma could be assumed to be the predisposing factor for this excessive fibrosis and dystrophic calcification in the spleen. PMID- 12762648 TI - Immunohistochemical study of the pancreatic basement membrane in non obese diabetic mice (NOD) with spontaneous autoimmune insulitis. AB - The role of the basement membrane as an antigenic structure in autoimmune diseases is controversial. To determine possible structural changes in the endocrine pancreatic basement membrane (PBM) in autoimmune diabetes, we studied the expression of laminin in the islets of 42 NOD mice, aged between 4 to 42 weeks, as an animal model of spontaneous diabetes. Insular lymphocytic inflammatory infiltration of variable intensity was present in 24 of these mice. An immunohistochemical staining using the streptavidin-biotin-peroxidase technique was performed on paraffin-embedded tissue sections, with a polyclonal antilaminin antibody. Staining for laminin was restricted to the basement membrane. In islets with no inflammatory infiltration, laminin was observed as a thin, continuous and uniform brown layer, covering the pericapsular basement membrane of the islets and their capillaries. The continuity of the PBM was lost in the islets with insulitis and the immunostaining showed clearcut interruption and destruction, particularly when the islets were in contact with inflammatory infiltrate. Our findings suggest that the loss of integrity of the PBM in islets with inflammatory infiltrate could facilitate antigenic exposure contributing towards the start o f autoimmune DM in NODmice. PMID- 12762649 TI - Evidence of apoptosis via TUNEL staining in muscle biopsy from patients with mitochondrial encephaloneuromyopathies. AB - Apoptosis is an evolution-conserved form of cell death essential for development and maintenance of tissue homeostasis. Dysregulation of apoptosis has been implicated in several pathological conditions, including neurodegenerative disorders. The crucial role of mitochondria in regulation of the apoptotic pathway prompted us to investigate the pattern of apoptosis in muscle biopsies from 17 patients with mitochondrial encephaloneuromyopathies caused by mtDNA defects. The results were compared with muscle biopsies from controls and from patients with myopathies without mitochondrial impairment. The terminal deoxynucleotidyl transferase-mediated dUTP nick and labelling (TUNEL) reaction was used as marker of apoptosis. Our findings were very heterogeneous, even between patients with the same mtDNA mutations, suggesting that tissue evaluation of apoptotic process is less useful than in vitro techniques, for investigating the role of apoptosis in mitochondrial pathologies. PMID- 12762651 TI - Elastofibroma associated with high grade leiomyosarcoma of the soft tissues: a light and ultrastructural study of one case. AB - Elastofibroma is a benign lesion occurring almost exclusively in the chest wall, parascapular region being the most frequent site. Rare lesions have been reported in other anatomic locations, but there are no reports about the co-existence of an elastofibroma with a malignant sarcoma. The purpose of the authors is to describe histologically and ultrastructurally the synchronous detection of an elastofibroma and a high grade leiomyosarcoma, speculating on eventual links between the two pathological states. PMID- 12762650 TI - Dexamethasone and fetal calf serum effects in differentiation of Vero cells cultured on type I collagen gel. AB - Vero cells, a fibroblastic cell line, were cultured on a collagen I gel without fetal calf serum (FCS), with the addition of 10% FCS, 10% FCS plus dexamethasone (DEX) or 20% FCS. Our objective was to determine the effects of DEX on the differentiation pattern of fibroblastic cells cultured on a collagen substrate. We found that cells cultured with or without 10% FCS were capable of migrating into the collagen matrix. Near the cells in the gel, we found the deposition of extracellular granulations. Cytochemical data suggests that this material is glycosaminoglycans and/or proteoglycans. Surrounding the cells, a fibronectin deposition was found in the collagen. Thus, these cells make up a structure similar to a loose connective tissue. On the other hand, cells cultured with 10% FCS plus DEX or with 20% FCS did not invade the collagen matrix but formed multiple cell layers poor in fibronectin. On collagen I gel surface, we found an acellular layer rich in collagen IV, which appeared between the cells and the substrate. Thus, DEX or 20% FCS, furnished to the cells cultured on a collagen gel, block cell migration into the substrate and induce them to produce a basement membrane-like structure. PMID- 12762652 TI - Oocyte development and egg envelope formation in Oreochromis niloticus, a mouth brooding cichlid fish. AB - The development of the oocyte and of its associated follicle cells in the Nile tilapia, Oreochromis niloticus, has been examined by optical and transmission electron microscopy. During oocyte development the female gamete of Orochromis niloticus increases in size because of the accumulation of yolk in its cytoplasm. As the accumulation of yolk proceeds, the organization of cortex of the oocyte becomes very complex; all of the cytoplasmic organelles and several populations of vesicles can be found. On the other hand follicle cells also undergo a series of modifications: they first become cuboidal then cylindrical and their cytoplasm become densely populated with organelles. The mature egg of Oreochromis niloticus is surrounded by a thin acellular envelope (chorion) assembled during oocyte development. Biochemical analysis of isolated and purified chorions from mature females was also performed. SDS-PAGE under reducing conditions showed a reproducible pattern of three major polypeptides (121, 66 and 50 kD), most of which being glycosylated. The pattern of synthesis and assembly of the egg envelope in Oreochromis niloticus, a mouth-brooding cichlid fish, is also discussed. PMID- 12762653 TI - Localisation and distribution of Wuchereria bancrofti antigens recognised by antisera from tropical pulmonary eosinophilia and from individuals with intestinal helminths. AB - Serological analyses of sera from patients with a typical picture of filarial tropical pulmonary eosinophilia (TPE) and sera from patients from a region non endemic for filariasis harbouring intestinal helminths, as Ascaris lumbricoides and Strongyloids stercoralis, revealed equally high titers of IgG4, usually considered diagnostic for filariasis. Ultrathin sections of adult worms of W. bancrofti embedded in the hydrophilic resin L.R. White were incubated with sera from patients with a typical picture of filarial tropical pulmonary eosinophilia (TPE) and sera from patients of a region that was non-endemic for filarial TPE but endemic for intestinal helminths. Both groups had a similar pattern of labelling, except that the labelling intensity was higher with the sera of patients with filarial TPE. The present study indicates relevant epitopes recognised by sera from TPE-patients and also individuals with intestinal helminths in all tissues of adult and intra-uterine microfilaria of W. bancrofti, instead of being localised in a specific nematode region. These findings suggest that people from areas not endemic for filariasis, but who harbour intestinal helminths, also share antifilarial antibodies in their serum that recognise antigens of adult worms and intrauterine microfilaria of W. bancrofti. PMID- 12762654 TI - The otoliths of the antarctic teleost Trematomus bernacchii: scanning electron microscopy and X-ray diffraction studies. AB - The authors studied the otoliths of the Nototheniid Trematomus bernacchii with scanning electron microscopy and X-ray diffraction analysis. Results obtained reveal that three otoliths are present: a large sagitta, a lapillus and a fragile asteriscus. Their sensorial faces appear finely decorated as shown by scanning electron microscopy (SEM). The sagitta and the lapillus are aragonitic while the asteriscus is vateritic, as demonstrated by X-ray diffraction. PMID- 12762655 TI - Morphology of the vas deferens of the budgerigar (Melopsittacus undulatus). AB - A morphological study of the budgerigar vas deferens was conducted to demonstrate the electron-microscopic features of its epithelial lining. The analysis showed that the vas deferens of the budgerigar was found to be of a tubular and serpentine structure, continuous with the epididymal region and lined with stereo ciliated pseudostratified epithelium, which contained folds projecting into the tubular lumen and a characteristic brush border. The epithelium consists of ciliated and non-ciliated cells with different electron densities. Ciliated cells were characterized by two morphologically distinct configurations: some cells were columnar and other ciliated cells were longer, thinner and dark. Non ciliated cells showed apical cytoplasmic expansions, which projected into the tubular lumen as protrusions. PMID- 12762656 TI - Ciliated fibroblasts in mouse connective tissues: a comparison between methods for estimating their frequency. AB - The frequency of ciliated fibroblasts in skin, gingiva, molar and incisor periodontal ligaments and incisor enamel-related periodontium of the mouse was estimated by straight counting or by methods based on the probability of observing a basal body in relation to other cell structures. Transmission electron microscopy of ultra-thin sections mounted in single slot grids was used. The results obtained with these methods differed, but indicated that periodontal ligament fibroblasts from molars or incisors generally had a higher ciliation index than the fibroblasts from skin and gingiva. These differences may not be real since the detection of cilia and/or centriolar structures seems to depend very much upon the plane of sectioning relative to the long axis of the fibroblasts, a situation which favours the more regularly arranged periodontal fibroblasts. This arrangement makes the periodontal tissues, particularly those of rodent incisors, a valuable model for studying ciliation in vivo because of the prompt response to experimental manipulation. PMID- 12762657 TI - Bacteriocyte population growth in Blattella germanica. AB - The number of bacteriocytes with nucleus in the M or S phase was analysed in relation to their rate of increase, throughout two nymphal instars (N1 and N6) of Blattella germanica Linnaeus (Blattaria, Blattellidae). We treated the experimental specimens with colcemid in order to visualize C-metaphases, and with labelled thymidine in order to evidence DNA synthesis in these bacteria-carrying cells. In both young and old nymphs, the C-metaphases showed a similar trend: their average number was only 0.3 per 100 bacteriocytes counted throughout the entire instars. In the young nymphs, the number of bacteriocytes in the S phase was congruent with the frequencies of the C metaphases. Since the number of M or S bacteriocytes was not sufficient to account for the observed increase in number of these endosymbiotic cells during nymphal development, we hypothesize a mechanism of bacterial transmission from bacteriocytes to other fat body cells that may explain the numerical growth of the bacteriocyte population. PMID- 12762658 TI - Catalytic asymmetric rearrangement of allylic N-aryl trifluoroacetimidates. A useful method for transforming prochiral allylic alcohols to chiral allylic amines. AB - [reaction: see text] A useful method for the conversion of prochiral allylic alcohols to chiral allylic amines of high enantiopurity is reported. N-(4 Methoxyphenyl)trifluoroacetimidates are excellent substrates for the palladium(II)-catalyzed allylic imidate rearrangement as the allylic trifluoroacetamide products can be deprotected in two steps to provide chiral nonracemic allylic amines. Di-mu-chlorobis[(eta(5)-(S)-(pR)-2-(2'-(4' isopropyl))oxazolinylcyclopentadienyl,1-C,3'-N))(eta(4) tetraphenylcyclobutadiene)cobalt]dipalladium (6a, COP-Cl) is a superior catalyst because it does not require activation with silver salts and provides rearranged allylic trifluoroacetamides in good yields and high enantiomeric purities. PMID- 12762659 TI - Highly enantioselective deracemization of linear and vaulted biaryl ligands. AB - [reaction: see text] A copper-mediated deracemization of the vaulted biaryl ligands VANOL and VAPOL can be readily achieved in the presence of (-)-spartiene. The optimal procedure involves the in situ generation of copper(II) and leads to the reproducible formation of (S)-VANOL and (S)-VAPOL in greater than 99% ee from the racemates. This method is superior to existing procedures for BINOL (92% ee). PMID- 12762660 TI - Preparation and characterization of alpha,alpha'-bisdiarylamino-capped oligothiophenes. AB - [reaction: see text] A series of alpha,alpha'-bisdiarylamino-capped oligothiophenes C(n) were prepared by the palladium-catalyzed reaction of the dibromo compounds A(i) with diarylamines, N,N-diarylamino-substituted thiophenes or 2,2'-bithiophenes BX(j). These easily oxidizable compounds exhibit a high tendency to form amorphous glasses and characteristic electrochemical and spectroscopic properties that depend significantly on the number of their thiophene moieties. PMID- 12762661 TI - Two-arm ferrocene amide compounds: synclinal conformations for selective sensing of dihydrogen phosphate ion. AB - [structure: see text] Ferrocene compounds bearing multiple amido groups selectively bind with the H(2)PO(4)(-) ion over other anions. The one-arm receptors form 1:1 complexes with the H(2)PO(4)(-) ion, whereas the two-arm receptors can accommodate two H(2)PO(4)(-) ions. The complexation modes and binding strengths are deduced from the NMR, calorimetry, fluorescence, and cyclic voltammetry studies. The two-arm ferrocene hexamide receptors likely exist in synclinal conformations to incorporate tetrahydrofuran molecules and H(2)PO(4)(-) ions. PMID- 12762663 TI - Zinc-coordination oligomers of phenanthrolinylporphyrins. AB - [structure: see text] We report the synthesis of mono- and bisphenanthrolinylporphyrins 5-(9-carboxy-1,10-phenanthroline-2),15-(p-tolyl) 2,8,12,18-tetraethyl-3,7,13,17-tetramethylporphyrin (MPPc) and 5,15-bis(9-carboxy 1,10-phenanthroline-2)-2,8,12,18-tetraethyl-3,7,13,17-tetramethylporphyrin (BPPc), respectively. The formation of dimers and oligomers, with MPPc and BPPc, respectively, upon exposure to zinc acetate is described. Oligomerization of the porphyrins was monitored spectrophotometrically, and a kinetic study was performed to estimate the average oligomer chain length. PMID- 12762662 TI - Highly selective synthesis of (E)-3-methyl-1-trialkylsilyl-3-en-1-ynes via trans selective alkynylation catalyzed by Cl2Pd(DPEphos) and stereospecific methylation with methylzincs catalyzed by Pd(tBu3P)2. AB - [reaction: see text] trans-Selective (>or=98%) monoalkynylation of 1,1-dibromo-1 alkenes and 1,1-dichloro-1-alkenes catalyzed by Cl(2)Pd(DPEphos) followed by stereospecific methylation with Me(2)Zn or MeZnX (X= Cl or Br) catalyzed by Pd((t)()Bu(3)P)(2) provides an efficient and stereoselective (>or=98%) route to 5, convertible to a wide variety of enynes and conjugated dienes. In the cases of 1,1-dibromo-1-alkenes, the Sonogashira alkynylation may also be satisfactory, but it is distinctly less satisfactory than the alkynylzinc reaction in cases where 1,1-dichloro-1-alkenes are used. PMID- 12762664 TI - Novel carbonyl allylation mediated by SnCl2/TiCl3 in water. AB - [reaction: see text] Under the Lewis acid catalysis offered by TiCl(3), SnCl(2) can efficiently mediate the aqueous Barbier reactions between aldehydes and allyl chloride or bromide. PMID- 12762666 TI - Sonogashira coupling reaction with diminished homocoupling. AB - [reaction: see text] The side product from homocoupling reaction of two terminal acetylenes in the Sonogashira reaction can be reduced to about 2% using an atmosphere of hydrogen gas diluted with nitrogen or argon. Terminal arylethynes, diarylethynes, and a few new arylpyridylethynes with donor substituents have been synthesized in very good yields. Comparative control experiments suggest that the homocoupling yield is determined by concentration of both catalyst and oxygen. PMID- 12762665 TI - Two syntheses of FF-MAS. AB - [reaction: see text] Follicular fluid-meiosis activating sterol (FF-MAS) has been shown to be an efficient inducer of meiotic maturation. It can potentially be used for improvements of in vitro fertilization techniques. Two short synthesis of FF-MAS are presented in this article. Both syntheses are based on microbiological degradations of sterol side chains. FF-MAS can be synthesized in nine steps from commercially available starting materials by both routes. PMID- 12762667 TI - Concise total synthesis of (+/-)-maritidine. AB - [reaction: see text] Maritidine can be readily obtained from the corresponding protected beta,gamma-unsaturated ketone. The quaternary carbon of maritidine was created for the first time via an intramolecular Heck reaction. PMID- 12762668 TI - Synthesis of cyclic proline-containing peptides via ring-closing metathesis. AB - [reaction: see text] Several dienes embedded in di- and tripeptides which incorporate proline have been prepared and subjected to ring-closing metathesis. Bicyclic peptides of well-defined amide geometry and of varying ring sizes were prepared. Several limitations of the cyclization step were revealed. PMID- 12762669 TI - Microwave-assisted solid-phase synthesis of phthalimides. AB - [reaction: see text] A traceless solid-phase synthesis of substituted phthalimides is proposed. The target compounds are obtained within minutes by a microwave-assisted cyclative cleavage in good yields and excellent purities. PMID- 12762670 TI - A stereoselective enyne cross metathesis. AB - [reaction: see text] Intermolecular enyne metathesis reaction of alkynes with olefins catalyzed by second-generation Grubbs catalyst (1) proceeded stereoselectively under ethylene atmosphere to produce 1,3-disubstituted butadienes with E stereochemistry. PMID- 12762671 TI - Asymmetric synthesis of alpha-amino allyl, benzyl, and propargyl silanes by metalation and rearrangement. AB - [reaction: see text] Metalation of a Boc-protected N-silylamine alpha to nitrogen results in migration of the silicon from nitrogen to carbon (reverse aza-Brook rearrangement), yielding an alpha-amino silane. The Boc group acts initially as a metalation-directing group and then to stabilize the nitrogen anion, providing a driving force for the rearrangement. In the presence of (-)-sparteine, the new chiral center is formed in >90% ee from allyl, benzyl, and propargylamines. PMID- 12762672 TI - Dibenzo[a,o]phenanthro[3,4-s]pycene, a configurationally stable double helicene: synthesis and determination of its conformation by NMR and GIAO calculations. AB - [reaction: see text] A double helicene formed by a pentahelicene and a heptahelicene with two rings in common was obtained by palladium-catalyzed cyclotrimerization of 3,4-didehydrophenanthrene and was characterized conformationally by NMR and GIAO calculations. PMID- 12762673 TI - Self-addition of a sterically hindered alkynylselenolate. AB - [structure: see text] The unprecedented formation of a complex macrocyclic selenium-carbon ring system, which is initiated by the action of coordinating reagents upon a sterically hindered alkynylselenolate, has been observed. The crystal structure of the product could be obtained and shows a rigid tricyclic arrangement consisting only of selenium and sp(2)-hybridized carbon atoms. This reactivity stands in contrast to the corresponding unsubstituted alkynylselenolates and is an example where a bulky substituent destabilizes an adjacent unsaturated pi-system. PMID- 12762674 TI - Stereoselective synthesis of 2-deoxy-beta-galactosides via 2-deoxy-2-bromo- and 2 deoxy-2-iodo-galactopyranosyl donors. AB - [reaction: see text] A series of 2-bromo- and 2-iodo-galactopyranosyl acetates and trichloroacetimidates were evaluated as glycosyl donors for the synthesis of 2-deoxygalactopyranosides. The best selectivity for the beta-glycosidic linkage was achieved by using 6-deoxy-3,4-carbonate-protected galactosyl donors. PMID- 12762675 TI - Stereoselective synthesis of functionalized precursors of the CDEF and CDE 2,6 dideoxy-tetra- and trisaccharide units of durhamycins A and B. AB - [structure: see text] Highly stereoselective syntheses of functionalized precursors of the CDEF and CDE 2,6-dideoxy-tetra- and trisaccharide units of the anti-HIV aureolic acids durhamycins A and B using 2-deoxy-2-iodo- and 2-deoxy-2 bromopyranosyl donors are described. PMID- 12762676 TI - Synthesis of bridged oligothiophenes: toward a new class of thiophene-based electroactive surfactants. AB - [structure: see text] A series of bridged oligothiophenes have been synthesized. Their novel molecular architecture, comprising two oligothiophenes linked together by a bridge at one alpha-extremity and one binding group at the other alpha-extremity, is expected to improve their utility as electroactive surfactants for semiconducting nanoparticles or organic electronics. PMID- 12762677 TI - Thiophene isosteres of 9,10-dithioanthraquinone. AB - [reaction: see text] The preparation of 1,3,5,7-tetramethyl-4,8-dihydrobenzo[1,2 c:4,5-c']dithiophene-4,8-dione and its conversion to the corresponding mono- and dithione are described. PMID- 12762678 TI - Synthesis of a [2]catenane around a Ru(diimine)3(2+) scaffold by ring-closing metathesis of olefins. AB - [reaction: see text] The synthesis of a ruthenium[2]catenane is described. One ring includes two 1,10-phenanthroline moieties, the other a bipyridinic unit. The interlocking ring system was formed by using a double ring closing metathesis reaction. Under irradiation, a rapid and selective decoordination of the bipyridinic fragment was observed, leading to a new catenane in which the metal is only coordinated to the bis-phenanthroline moiety. PMID- 12762679 TI - Enantioselective total synthesis of aspidophytine. AB - [structure: see text] An enantioselective total synthesis of aspidophytine is described. The indole fragment bearing a cis-alkene substituent was efficiently prepared through radical cyclization of a 2-alkenylphenylisocyanide followed by Sonogashira coupling of the generated 2-iodoindole derivative with a functionalized acetylene unit. After formation of the 11-membered cyclic amine, the aspidosperma skeleton and lactone ring were constructed to complete the total synthesis. PMID- 12762681 TI - Directed ortho metalation approach to C-7-substituted indoles. Suzuki-Miyaura cross coupling and the synthesis of pyrrolophenanthridone alkaloids. AB - [reaction: see text] Although the indole N-phosphinoyl derivative 4 undergoes n BuLi deprotonation/electrophile quench to afford C-7-substituted products, its deprotection requires harsh conditions. On the other hand, the N-amide 12, upon sequential or one-pot C-2 metalation, silylation, C-7 metalation, and electrophile treatment, furnishes indoles 7 in good overall yields. In combination with the Suzuki-Miyaura protocol, C-7 aryl (heteroaryl)-substituted indoles 14 and 16 are obtained, including hippadine and pratosine, members of the pyrrolophenanthridone alkaloid family. PMID- 12762680 TI - Ruthenium-catalyzed vinylsilane synthesis and cross-coupling as a selective approach to alkenes: benzyldimethylsilyl as a robust vinylmetal functionality. AB - [reaction: see text] Ruthenium-catalyzed alkyne hydrosilylation or silyl-alkyne Alder ene reactions provide entry into benzyldimethylsilyl (BDMS)-substituted alkenes. The BDMS-vinylsilanes are further elaborated through mild palladium catalyzed cross coupling and show significant stability to intervening synthetic operations, including silyl ether deprotection. PMID- 12762682 TI - Ionic disparity of identical molecules in polymorphs. AB - [structure: see text] Formation of ionic pairs of identical molecules, extremely common in Nature but exceptional for diffractometric observations in the crystalline state, has been found in one of two polymorphs of a nucleobase analogue, 4,6-pyrimidinedione (1). The occurrence of the neutral and ionic polymorphs of 1 in many respects resembles neutral-ionic transitions of electon donor/acceptor complexes. The ionization affects the reactivity of 1 and illustrates general properties of H-tautomers with low-disproportionation energy. PMID- 12762683 TI - "Magic rod" rotaxanes: the hydrogen bond-directed synthesis of molecular shuttles under thermodynamic control. AB - [reaction: see text] Peptide [2]- and [3]rotaxanes are assembled in high yields under thermodynamic control using hydrogen bonding interactions and reversible cross olefin metathesis. PMID- 12762684 TI - Molecular folding of C60 acetylenic cyclophanes: pi-stacking of superimposed aromatic rings. AB - [structure: see text] The syntheses of two distinct families of phenylyne helical cyclophanes with potential for organic materials are described. The meta-bonded atropisomers afford interesting bowtie-like and butterfly-like conformers from a palladium(0), copper-mediated coupling sequence. Molecular modeling revealed the contrasting stereochemistry in these systems from differential molecular folding pathways during cyclization. The interplanar separation of the superimposed aromatic rings is approximately 3.5 A. PMID- 12762685 TI - Palladium-catalyzed alpha-arylation of azlactones to form quaternary amino acid derivatives. AB - [reaction: see text] The synthesis of alpha-aryl-alpha-alkyl amino acid derivatives from alpha-amino acids by the arylation of azlactone derivatives is reported. Arylation of azlactones derived from alanine, valine, phenalanine, phenyl glycine, and leucine all provided good yields of the arylated product. Mechanistic studies of this reaction revealed that a stable complex containing a ligand formed by reaction of dba with the azlactone accounts for a new inhibiting effect of dba when reactions are initiated with Pd(dba)(2). PMID- 12762686 TI - Selectivities in reactions of organolithium reagents with unprotected 2 halobenzoic acids. AB - [reaction: see text] Exposing 2-fluorobenzoic acid (1a) to 2.2 equiv of LTMP at ca. -78 degrees C leads to deprotonation at the 3-position whereas 2 chloro/bromobenzoic acids (1b,c) are lithiated adjacent to the carboxylate. The resulting dianions 3Li-1a and 6Li-1b,c are trapped as such by chlorotrimethylsilane. In the absence of internal quench, 6Li-1b,c isomerize to the more stable 3Li-1b,c. The latter eliminate lithium halide and set free benzyne-3-carboxylate (2) that reacts regioselectively with LTMP to give 3 tetramethylpiperidinobenzoic acid (3). PMID- 12762687 TI - Synthesis of the [5-7-6] tricyclic core of guanacastepene A via an intramolecular Mukaiyama aldol reaction. AB - [reaction: see text] A synthesis of the tricyclic [5-7-6] skeleton of guanacastepene A is described. The six-membered ring of guanacastepene A was constructed by a Diels-Alder reaction. After several functional group transformations, it was coupled to beta-iodocyclopentenone. Lithium dimethylcuprate conjugate addition followed by an intramolecular Mukaiyama aldol reaction furnished the desired [5-7-6] tricyclic ring system. PMID- 12762688 TI - Asymmetric synthesis of (+)-L-733, 060 and (+)-CP-99, 994 based on a new chiral 3 piperidinol synthon. AB - [reaction: see text] Selective and potent neurokinin substance P receptor antagonists (+)-L-733, 060 (1) and (+)-CP-99, 994 (2) have been synthesized starting from a new (3S)-piperidinol synthon derived from l-glutamic acid. The methods featured a C-2 regioselective reduction of glutarimide (9), Lewis acid promoted Si to C-2 phenyl group migration of 10, and stereoselective reduction of acetylated oxime 19 as the key steps. PMID- 12762689 TI - Synthesis of 2,5-disubstituted tetrahydrofurans by stereospecific elimination cyclization of 1-iodomethyl-1,5-bis-epoxides. AB - [reaction: see text] Treatment of 1-iodomethyl-1,5-bis-epoxides with zinc in refluxing ethanol affords cis or trans 2-vinyl-5-(1-hydroxyethy)-substituted tetrahydrofurans stereospecifically in high yield. PMID- 12762690 TI - Unified synthesis of eudesmanolides, combining biomimetic strategies with homogeneous catalysis and free-radical chemistry. AB - [reaction: see text] A general procedure for the synthesis of both 12,6- and 12,8 eudesmanolides has been developed. The key step is the titanocene-catalyzed radical cyclization of accessible epoxygermacrolides. The novel reagent 2,4,6 trimethyl-1-trimethylsilylpyridinium chloride, both compatible with oxiranes and capable of regenerating Cp(2)TiCl(2) from Cp(2)Ti(Cl)H and Cp(2)Ti(Cl)OAc, played an important role in the catalytic cycle leading to exocyclic alkenes. PMID- 12762691 TI - Unprecedented conversion of triethylamine and disulfur dichloride into a thienopentathiepin and a heptathiocane. AB - [reaction: see text] In a remarkable cascade reaction, triethylamine is converted into the thienopentathiepin 2a and the heptathiocane 3a by a preequilibrated solution of disulfur dichloride and DABCO in chloroform. PMID- 12762692 TI - Stereoselective reactions of a (-)-quinic acid-derived enone: application to the synthesis of the core of scyphostatin. AB - [reaction: see text] A (-)-quinic acid-derived enone, with the trans-1,2-diol protected as a 2,3-dimethoxybutanediyldioxy ketal, provides an excellent template for further highly stereoselective elaboration as exemplified by its conversion into the core of scyphostatin, a potent inhibitor of neutral sphingomyelinase. PMID- 12762693 TI - Sequential one-pot glycosylations using 1-hydroxyl and 1-thiodonors. AB - [reaction: see text] A novel sequential glycosylation procedure is described that combines the use of 1-hydroxyl and thiodonors. The Ph(2)SO/Tf(2)O-mediated dehydrative condensation of 1-hydroxyl donors with thioglycosides affords in good yield the thiodisaccharides, which in turn can be activated by the same activator system to furnish trisaccharides. The alpha-Gal epitope and a hyaluronan trisaccharide were efficiently assembled in a one-pot procedure. PMID- 12762694 TI - Multivalent neoglycoconjugates by regiospecific cycloaddition of alkynes and azides using organic-soluble copper catalysts. AB - [reaction: see text] The construction of multivalent neoglycoconjugates is efficiently achieved by the regiospecific catalytic cycloaddition of alkynes and azides using the organic-soluble copper complexes (Ph(3)P)(3).CuBr and (EtO)(3)P.CuI. The simultaneous use of microwave irradiation shortened notably the reaction times. PMID- 12762695 TI - First zipper-featured molecular duplexes driven by cooperative donor-acceptor interaction. AB - [structure: see text] The first class of zipper-shaped artificial duplexes, which are driven by multiple donor-acceptor interactions between electron-rich 1,5 dioxynaphthalene or 1,4-dioxybenzene and electron-deficient pyromellitic dimide units, have been studied in organic media by (1)H NMR, UV-vis, and vapor pressure osmometry. (1)H NMR binding investigations reveal substantial cooperativity of the donor-acceptor interaction in the duplexes. PMID- 12762696 TI - Enantioselective syntheses of cryptocarya triacetate, cryptocaryolone, and cryptocaryolone diacetate. AB - [reaction: see text] The enantioselective syntheses of three natural products from Cryptocarya latifolia have been achieved in 13-15 steps from ethyl sorbate. The route relies upon an enantio- and regioselective Sharpless dihydroxylation and a palladium-catalyzed reduction to establish the absolute stereochemistry. The route also relies upon a highly (E)-selective olefin cross-metathesis reaction to form trans-delta-hydroxy-1-enoates. The resulting delta-hydroxy-1 enoates were subsequently converted into cryptocarya triacetate, cryptocaryolone, and cryptocaryolone diacetate. PMID- 12762697 TI - Toward the synthesis of reidispongiolide a: stereocontrolled synthesis of the C17 C22 and C23-C35 degradation fragments. AB - [structure: see text] By relying on the asymmetric aldol reactions of chiral ketones, a highly stereocontrolled synthesis of each of the C(17)-C(22) and C(23) C(35) degradation fragments of reidispongiolide A has been achieved. This permits a configurational assignment of the complete C(17)-C(36) region of this antimitotic macrolide, along with providing advanced intermediates for a projected total synthesis. PMID- 12762699 TI - Catalytic enantioselective desymmetrization of COT-monoepoxide. Maximum deviation from coplanarity for an S(N)2'-cuprate alkylation. AB - [reaction: see text] The first alkylative and enantioselective ring-opening of COT-monoepoxide (1) without the occurrence of any ring-contraction-isomerization by the use of in situ-formed organocuprates is reported. Because of the particular geometric constraint of compound 1, this work reports the largest deviation from coplanarity between the pi-orbital of the double bond and the sigma-bond connecting the leaving group ever observed for an S(N)2'-cuprate alkylation. PMID- 12762698 TI - Relative rates of Michael reactions of 2'-(phenethyl)thiol with vinyl sulfones, vinyl sulfonate esters, and vinyl sulfonamides relevant to vinyl sulfonyl cysteine protease inhibitors. AB - [reaction: see text] The relative rates of Michael additions of 2' (phenethyl)thiol to representative vinyl sulfonyl Michael acceptors were measured. The dependence of the reactivity of the Michael acceptor on the nature of the sulfonyl R substituent was determined in order to evaluate the effect of these substituents on the inactivation kinetics of comparably substituted vinyl sulfonyl cysteine protease inhibitors. The rates of these Michael additions vary over 3 orders of magnitude, with phenyl vinyl sulfonate esters (R = OPh) being ca. 3000-fold more reactive than N-benzyl vinyl sulfonamides (R = NHBn). PMID- 12762700 TI - Synthesis of fused oxabicyclic systems by metal-catalyzed intramolecular addition of 1,3-cycloalkyldiones to alkynes. AB - [reaction: see text] Readily available 4-propargyl-1,3-cyclohexanediones and cyclopentanediones can be chemo- and regioselectively cycloisomerized to synthetically appealing fused oxabicyclic systems by simply stirring at room temperature with catalytic amounts of an appropriate metal complex. PMID- 12762701 TI - Iron(III)-catalyzed Prins-type cyclization using homopropargylic alcohol: a method for the synthesis of 2-alkyl-4-halo-5,6-dihydro-2H-pyrans. AB - [reaction: see text] A new Prins-type cyclization between homopropargylic alcohol and aldehydes in the presence of FeX(3) to obtain 2-alkyl-4-halo-5,6-dihydro-2H pyrans in good yield is described. Osmium-catalyzed cis dihydroxylation provided direct access to trans-2-alkyl-3-hydroxy-tetrahydro-pyran-4-ones. Anhydrous ferric halides are also shown to be excellent catalysts for the standard Prins cyclization using homoallylic alcohol. Isolation of an intermediate acetal provides substantiation of a proposed mechanism. PMID- 12762702 TI - Modular, parallel synthesis of an illudinoid combinatorial library. AB - [structure: see text] A library of 49 illudinoids was prepared via a three-step synthesis using a parallel synthesizer and solid-phase extractive purification. Products could be rapidly prepared for initial biological testing against three cancer cell lines, which revealed library members that inhibit nonsmall cell lung cancer. The activities of the library members are not easily correlated with structure, meaning the empirical approach used here is essential for such nonintuitive discoveries. PMID- 12762703 TI - Synthesis and incorporation into PNA of fluorinated olefinic PNA (F-OPA) monomers. AB - [structure: see text] A fluorinated OPA monomer containing the base thymine ((Z) t-F-OPA) was synthesized in 12 steps, featuring a highly selective allylic over homoallylic Mitsunobu substitution for the introduction of the nucleobase. F-OPA modified PNA decamers were prepared by the MMTr/acyl protection strategy. The thermal stability of duplexes of PNA decamers containing (Z)-t-F-OPA units with antiparallel complementary DNA was measured. We found a strong dependence of stability from the sequential position of the (Z)-t-F-OPA units, ranging from DeltaT(m) of +2.4 to -8.1 degrees C/modification relative to unmodified PNA. PMID- 12762704 TI - Organosilanes in synthesis: application to an enantioselective synthesis of methyl-l-callipeltose. AB - [reaction: see text] Methyl-l-callipeltose, the carbohydrate associated with callipeltoside A, has been synthesized in eight steps and 23% overall yield from enantioenriched allylsilane 6 and acetaldehyde. The key steps are a highly diastereoselective formal [4 + 2] annulation and a Cr(VI)-catalyzed oxidative C-C bond cleavage to produce lactone 11. PMID- 12762705 TI - Total synthesis of (+)-strictifolione. AB - [reaction: see text] The synthesis of (+)-strictifolione was achieved from 3 phenylproprionaldehyde by using enantioselective allyltitanations to control the stereogenic centers at C6, C4', and C6' and a cross-methathesis to control the configuration of the double bond at C1'-C2'. PMID- 12762706 TI - The first method for protection-deprotection of the indole 2,3-pi bond. AB - [reaction: see text] The scope and generality of a new reaction of indoles with MTAD is discussed. In most cases the ene-type reaction proceeds within seconds or minutes at 0 degrees C to provide the urazole adducts in high yield. This reaction provides the first method for protecting the indole 2,3-double bond since the urazole adducts can be reconverted to the starting indole (retro-ene) simply by heating. PMID- 12762707 TI - One-pot, two-step, practical catalytic synthesis of 2,5-diformylfuran from fructose. AB - [reaction: see text] A practical, one-pot, two-step catalytic method is described for the synthesis of 2,5-diformylfuran (DFF) from fructose via dehydration to 5 (hydroxymethyl)furfural (HMF), followed by in situ catalytic air-oxidation. PMID- 12762708 TI - Selective domino ring-closing metathesis-cross-metathesis reactions between enynes and electron-deficient alkenes. AB - [reaction: see text] A selective domino ring-closing metathesis (RCM)-cross metathesis (CM) process between enynes and electron-deficient alkenes is reported. The conditions have been optimized for enynes 3 and methyl acrylate with catalyst IV. The scope and limitations of this reaction are described, and a possible mechanism is discussed. PMID- 12762709 TI - Oxidative demetalation of cyclohexadienyl ruthenium(II) complexes: a net Ru mediated dearomatization. AB - [reaction: see text] An experimentally simple method for the demetalation of spirocyclic cyclohexadienylruthenium(II) complexes has been developed. Treatment of an alkoxy-substituted cyclohexadienyl complex with CuCl(2) affords either azaspiro[4.5]decane derivatives or heavily functionalized tetrahydroisoquinolines. The former reaction manifold completes a net Ru-mediated dearomatization as the organometallic starting materials are prepared from (eta(6)-arene)Ru(II) precursors. Both of these heterocyclic products are well suited for further synthetic elaboration. PMID- 12762711 TI - Application of physical organic methods to the investigation of organometallic reaction mechanisms. AB - For the past 75 years, physical organic chemistry has been the foundation for the elucidation of mechanisms, and the understanding of structure/activity relationships, in organic chemistry. The rational design of experiments and the critical analysis of data have been the hallmarks of this field. In this Perspective, we revisit some of the studies carried out during the past 30 years in which our group has used the tools of physical organic chemistry to investigate the mechanisms of a variety of organometallic reactions. The Perspective covers a broad spectrum of metal-mediated reactions including organic reductions, alkene, carbon monoxide and nitric oxide migratory insertion reactions, C-H bond activation, and the reactions of metal-heteroatom bonds with organic molecules. These studies, along with similar ones carried out in other groups, have demonstrated that the principles and methods of physical organic chemistry have adapted well to the study of organometallic reaction mechanisms. The understanding obtained in this way has aided the development of useful metal mediated reactions in a wide range of areas, especially in industrial catalysis and in applications to problems in organic synthesis. PMID- 12762713 TI - Direct enantioselective Michael addition of aldehydes to vinyl ketones catalyzed by chiral amines. AB - Chiral amines such as (S)-2-[bis(3,5-dimethylphenyl)methyl]pyrrolidine and the C(2)-symmetric (2S,5S)-2,5-diphenylpyrrolidine can catalyze the direct enantioselective Michael addition of simple aldehydes to vinyl ketones. The conditions for this organocatalytic reaction have been optimized and it has been found that the chiral amines catalyze the formation of optically active substituted 5-keto aldehydes in good yields and enantioselectivities, using aldehydes and, e.g., methyl vinyl ketone as starting compounds. Taking into account that the chiral amine can activate the aldehyde and/or the enone, the mechanism for the reaction has been investigated. On the basis of intermediate synthesis, nonlinear effect, and theoretical investigations, the mechanism for the catalytic direct enantioselective Michael addition of aldehydes to vinyl ketones is discussed. PMID- 12762712 TI - Hetaryleneaminopolyols and hetarylenecarbopeptoids: a new type of glyco- and peptidomimetics. Syntheses and studies on solution conformation and dynamics. AB - Ready access to a new class of oligomers has been demonstrated by the synthesis of hetaryleneaminopolyols and hetarylenecarbopeptoids using 3-hydroxymethyl-5-(4 amino-4-deoxy-d-arabinotetritol-1-yl)-2-methylfuran and 5-(4-amino-4-deoxy-d arabinotetritol-1-yl)-2-methyl-3-furoic acid as novel scaffolds. The conformational behavior of peptidomimetics 22, 23, 25, 26, and 36 have been analyzed by NMR spectroscopy and extensive molecular dynamics simulations. MD simulations using the GB/SA continuum solvent model for water and the MM3 force field provide a population distribution of conformers which satisfactorily agrees with the experimental NMR data for the torsional degrees of freedom of the molecule. PMID- 12762714 TI - Digital processing with a three-state molecular switch. AB - Certain molecular switches respond to input stimulations producing detectable outputs. The interplay of these signals can be exploited to reproduce basic logic operations at the molecular level. The transition from simple logic gates to complex digital circuits requires the design of chemical systems able to process multiple inputs and outputs. We have identified a three-state molecular switch that responds to one chemical and two optical inputs producing two optical outputs. We have encoded binary digits in its inputs and outputs applying positive logic conventions and demonstrated that this chemical system converts three-digit input strings into two-digit output strings. The logic function executed by the three-state molecular switch is equivalent to that of a combinational logic circuit integrating two AND, two NOT, and one OR gate. The three states of the molecular switch are a colorless spiropyran, a purple trans merocyanine, and its yellow-green protonated form. We have elucidated their structures by X-ray crystallography, (1)H NMR spectroscopy, COSY and NOE experiments, as well as density functional calculations. The three input stimulations controlling the interconversion of the three states of the molecular switch are ultraviolet light, visible light, and H(+). The two outputs are the absorption bands in the visible region of the two colored states of the molecular switch. We have monitored the switching processes and quantified the associated thermodynamic and kinetic parameters with the aid of (1)H NMR and visible absorption spectroscopies. PMID- 12762715 TI - Convergent synthesis of polyhalogenated quinoline C-nucleosides as potential antiviral agents. AB - 2,5,6-Trichloro-1-(beta-d-ribofuranosyl)benzimidazole (TCRB) and 2-bromo-5,6 dichloro-1-(beta-d-ribofuranosyl)benzimidazole (BDCRB) are benzimidazole nucleosides that exhibit strong and selective anti-HCMV activity. We proposed to synthesize 2-halo-6,7-dichloro-4-(beta-d-ribofuranosyl)quinolines as 6 + 6 bicyclic analogues of TCRB. The synthesis used Wittig reactions in two key steps. The first Wittig reaction coupled a fully functionalized benzene with a ribofuranose derivative to provide (Z)-6-O-(tert-butyldimethylsilyl)-1-(4,5 dichloro-2-nitrophenyl)-1,2-dideoxy-3,4-O-isopropylidene-d-allo-1-enitol (5) as the basic skeleton for the target compounds. The following electrophile-mediated intramolecular cyclization of the cis-alkene (5) was found to afford (1S,2S)-2,5 anhydro-1-bromo-6-O-(tert-butyldimethylsilyl)-1-deoxy-1-(4,5-dichloro-2 nitrophenyl)-3,4-O-isopropylidene-d-allitol (8) as the major product. This alpha stereoselectivity was contrary to the literature precedence. A double-bond isomerization was established to be the cause of the unexpected stereochemistry. The bromo group of 8 was displaced by a hydroxyl group. Oxidation of the hydroxy group and the reduction of a phenylnitro group provided (2S)-1-(2-amino-4,5 dichlorophenyl)-2,5-anhydro-6-O-(tert-butyldimethylsilyl)-3,4-O-isopropylidene-d allose (11), which was subjected to the second Wittig reaction with a phosphacumulene to construct 4-[5-O-(tert-butyldimethylsilyl)-2,3-O isopropylidene-alpha-d-ribofuranosyl]-6,7-dichloroquinolin-2-one (13). Halogenation followed by deprotection of 13 and led to the synthesis of 4-(alpha d-ribofuranosyl)-2,6,7-trichloroquinoline (17) as the major product. The 2 aminophenone alpha-nucleoside (11) was successfully anomerized to the beta-anomer (19), which led to the synthesis of the targeted 2-chloro- and 2-bromo-6,7 dichloro-4-(beta-d-ribofuranosyl)quinolines (18and 21, respectively). PMID- 12762717 TI - A novel anthracenyl tagged protecting group for "phase-switching" applications in parallel synthesis. AB - A new "phase-switching" protecting group 1 that facilitates the parallel synthesis of carboxylic acids, esters, and carboxamides is described. Its use permits chemistries to be performed in solution, which may be conveniently monitored with conventional analytical techniques, followed by subsequent immobilization onto a solid-phase support to aid compound purification. Carboxylic acids, esters, and carboxamides are then cleaved from the solid support following activation of the "safety-catch" and treatment with the desired nucleophile. PMID- 12762716 TI - Synthesis of 5,12-dioxocyclam nickel (II) complexes having quinoxaline substituents at the 6 and 13 positions as potential DNA bis-intercalating and cleaving agents. AB - Several dioxocyclams containing quinoxaline moieties, as well as their nickel(II) complexes were synthesized and studied for their ability to bind and oxidatively cleave DNA. Although no evidence for binding by intercalation was found, the ability of the Ni(II) complexes to cleave DNA in the presence of Oxone was strongly dependent on both the nature and the spatial orientation of the quinoxaline moieties, suggesting at least transient association of these complexes with DNA. PMID- 12762718 TI - A tandem metal carbene cyclization-cycloaddition approach to the pseudolaric acids. AB - An approach toward the synthesis of the antifungal and cytotoxic pseudolaric acids based on the tandem metal carbene cyclization-cycloaddition reaction is described. Using this strategy, the advanced intermediate 3a bearing three of the four stereocenters of the target molecules has been synthesized. The substrate controlled diastereoselectivity of the tandem carbene cyclization-cycloaddition was preferential for the undesired diastereomer, but reagent control through the use of Hashimoto's chiral rhodium catalyst Rh(2)(S-BPTV)(4) reversed the selectivity in favor of 3a. Ring opening of the oxabicyclic nucleus to give a hydroxycycloheptene has been demonstrated in a model study. PMID- 12762719 TI - Studies on intramolecular Diels-Alder reactions of furo[3,4-c]pyridines in the synthesis of conformationally restricted analogues of nicotine and anabasine. AB - En route to conformationally restricted analogues of nicotine and anabasine, a novel synthetic route to bridged anabasines is described that hinges on a domino intramolecular [4 + 2]-cycloaddition/ring opening-elimination sequence of 3-amino substituted furo[3,4-c]pyridines. Extension of this route to bridged nicotines, however, proved abortive, even when the dienophile tether is activated by a p tolylsulfonyl group or when the diene moiety is activated by an electron releasing methoxy substituent. A detailed density functional theoretical study (B3LYP/6-31+G) was undertaken to provide insight into the factors that facilitate an intramolecular Diels-Alder reaction in the former case. PMID- 12762720 TI - Enantioselective total synthesis of the antitumor macrolide rhizoxin D. AB - The convergent, highly enantioselective synthesis of rhizoxin D, a natural product possessing potent antitumor and antifungal bioactivity, is described. The C(1)-C(9) fragment of the molecule was synthesized utilizing a threefold pseudosymmetric intermediate ultimately derived from gamma-butyrolactone. The central core of rhizoxin D was prepared via a chiral resolution/asymmetric aldol protocol. Several methods for the generation of the polyene fragment were explored, and the side-chain was ultimately prepared from serine in six steps. The unification of the left and right wings of the molecule was achieved using a one-step olefination protocol, and the macrocyclization was carried out using a Horner-Emmons olefination at the C(2)-C(3) olefin. PMID- 12762721 TI - Hydroxy alkenenitriles: diastereoselective conjugate addition-alkylations. AB - Chelation-controlled conjugate addition of Grignard reagents to cyclic and acyclic gamma-hydroxyalkenenitriles stereoselectively generates beta-substituted hydroxynitriles. t-BuMgCl-induced deprotonation of gamma-hydroxyalkenenitriles followed by chloride-alkyl exchange from a second Grignard reagent, generates an alkylmagnesium alkoxide that triggers conjugate addition. Alkylation of the resulting magnesiated nitrile with alkyl halide and carbonyl electrophiles efficiently installs two new bonds and up to three stereocenters in a single synthetic operation. PMID- 12762722 TI - A novel synthesis of functionalized tetrahydrofurans by an Oxa-Michael/Michael cyclization of gamma-hydroxyenones. AB - An approach to highly functionalized tetrahydrofuran derivatives based upon a novel Oxa-Michael/Michael dimerization of cis-gamma-hydroxyenones is presented. The reaction begins with either 1,2-dioxines or trans-gamma-hydroxyenones and proceeds by addition of one molecule of trans-gamma-hydroxyenone to another molecule of cis- or trans-gamma-hydroxyenone catalyzed by an alkoxide or hydroxide base. Subsequent intramolecular Michael addition of the keto-enolate gives the observed tetrahydrofurans. Substitution at both the 2- and 4-positions of the gamma-hydroxyenone is tolerated; however, for 4-substituted gamma hydroxyenones, selectivity issues arise due to the possibility of heterochiral or homochiral dimerizations. The major products were those with all contiguous groups trans. PMID- 12762723 TI - The surprisingly facile thermal dehalogenation of chlorinated aromatics by a hydroaromatic donor solvent. Tautomerization of chlorinated phenols and anilines. AB - Up to 600 K, chlorinated benzenes and naphthalenes are inert in a hydrogen atom donating solvent such as 9,10-dihydroanthracene. However, when a hydroxyl or amine group is attached to the 2 or 4 position relative to chlorine, a surprisingly facile and selective hydrodehalogenation occurs at temperatures between 530 and 630 K. These features are the result of the onset of tautomeric equilibria for the chlorophenols or -anilines, creating the corresponding enones or imines, respectively, as reactive intermediates. The mechanism of dehalogenation involves the rate-determining transfer of a hydrogen atom from the solvent to the reactive intermediate according to a reverse radical disproportionation (RRD), followed by elimination of chlorine. The reactivity of mono- and dichlorophenols, chloroanilines, 4-chloro-1-naphthol, and 4-chloro-1 naphthylamine dissolved in 9,10-dihydroanthracene has been investigated. By means of density functional theory (DFT) calculations, the energies for the tautomers and their hydrogen affinities have been established. The experimental selectivities could be adequately reproduced by the computed data. PMID- 12762724 TI - A solid-state nitrogen-15 NMR and ab initio study of nitrobenzenes. AB - Insight into the unexpectedly small range of isotropic nitrogen chemical shifts in nitrobenzene derivatives is gained through measurements of the chemical shift (CS) tensor by solid-state NMR experiments and ab initio molecular orbital (MO) and density functional theory (DFT) calculations. The principal components, delta(ii), of the (15)N CS tensors have been measured for nitrobenzene, 4 nitroaniline, 4-nitrotoluene, 4-nitroanisole, 4-nitroacetophenone, nitromesitylene, and 2,4,6-tri-tert-butylnitrobenzene. No obvious correlations of the delta(ii) values with traditional reactivity parameters were observed. The CS tensor components change significantly for the para-substituted nitrobenzenes, but these variations nearly cancel to yield isotropic shifts that fall in a range of only 3 ppm. Ab initio calculations of the delta(ii) values at the HF level are in poor agreement with the experimental values, whereas MP2 calculations and DFT calculations employing the B3LYP functional are in better agreement with experiment. The calculated (B3LYP/6-311G) delta(ii) values follow a trend in which delta(11) and delta(33) increase while delta(22) decreases with the accepted electron withdrawing ability of the para substituent. These changes tend to cancel yielding a variation in delta(iso) of only 4 ppm. These calculations indicate that the CS tensor has the same orientation as the carbon CS tensor in the isoelectronic benzoate anion: delta(11) bisects the O-N-O angle, delta(33) is perpendicular to the NO(2) plane, and delta(22) is in the NO(2) plane and perpendicular to delta(11). PMID- 12762725 TI - Density functional study on the mechanism of the vanadium-catalyzed oxidation of sulfides by hydrogen peroxide. AB - A computational study with the Becke3LYP method is carried out on the mechanism of the reaction of complexes V(O)(L)(OOH) and V(O)(LH)(OO) (L = O(CH)(3)N(CH(2))(2)O) with CH(3)S-SCH(3), a system that stands as a model for experimental systems where the metal complex contains larger chelating Schiff bases and the substrate is bis(tert-butyl) disulfide. The different possible isomers of both the hydroperoxo V(O)(L)(OOH) and the peroxo V(O)(LH)(OO) forms of the catalyst are explored, and the reactivity of the most stable among them with the dimethyl disulfide substrate is studied through location of the corresponding transition states. A large variety of reactive paths happen to exist, though in all cases the reaction takes place through a direct transfer process, with the simultaneous formation of the S-O bond and breaking of the O-O bond being the rate-limiting step. PMID- 12762726 TI - Independent generation and study of 5,6-Dihydro-2'-deoxyuridin-6-yl, a member of the major family of reactive intermediates formed in DNA from the effects of gamma-radiolysis. AB - Nucleobase radicals are the major family of reactive intermediates formed when nucleic acids are exposed to gamma-radiolysis. Elucidation of their reactivity is complicated by the formation of multiple species randomly throughout the biopolymers. 5,6-Dihydro-2'-deoxyuridin-6-yl (1) was generated upon photolysis (350 nm) of the respective tert-butyl ketone (2). The radical abstracts hydrogen atoms from beta-mercaptoethanol (k = 8.8 +/- 0.5 x 10(6) M(-)(1) s(-)(1)) and 2,5 dimethyltetrahydrofuran (k = 31 +/- 2.5 M(-)(1) s(-)(1)). The latter was used as a model for the 2-deoxyribose component of DNA. The major product formed in the presence of O(2) was 6-hydroxy-5,6-dihydro-2'-deoxyuridine (11), which is believed to be formed directly from the peroxy precursor and not via elimination of superoxide. Small amounts of 2-deoxyribonolactone (13) were also formed under aerobic conditions. This product is believed to result from intramolecular hydrogen atom abstraction by the C6-peroxyl radical (14) and suggests that gamma radiolysis may indirectly result in oxidation of the C1'-position of nucleotides, despite the inaccessibility of this hydrogen in duplex DNA. PMID- 12762727 TI - Significantly enhanced reactivities of the nucleophilic substitution reactions in ionic liquid. AB - We have investigated the reactivities of various metal fluorides in the nucleophilic fluorination of 2-(3-methanesulfonyloxypropyl)naphthalene (1) as a model compound in the presence of 1-n-butyl-3-methylimidazolium tetrafluoroborate ([bmim][BF(4)]). The higher periodic alkali metal fluorides demonstrate greater reactivity. The fluorination using CsF among the alkali metal fluorides was completed in 20 min, affording the desired product 2-(3-fluoropropyl)naphthalene (2a, 95%) without any byproducts. However, the fluorinations using alkali earth, transition, and low periodic alkali metal fluorides under the same conditions occurred rarely or not at all. We have also carried out the various facile nucleophilic substitutions such as halogenations, acetoxylation, nitrilation, and alkoxylations of mesyloxyalkane 1 and 2-(3-bromopropyl)naphthalene (6) at the primary aliphatic position using the potassium halides, acetate, cyanide, and alkoxides, respectively, in the presence of ionic liquids. These reactions provided the desired products, such as 2-(3-halopropyl)naphthalenes 5-7 (95% for Cl, 96% for Br, and 93% for I), 2-(3-acetoxypropyl)naphthalene (8, 95%), 2-(3 cyanopropyl)naphthalene (9, 93%), and 2-(3-methoxypropyl)naphthalene (10, 92%). PMID- 12762728 TI - Development of a [3+3] cycloaddition strategy toward functionalized piperidines. AB - This paper describes a novel route to functionalized piperidines via a formal [3+3] cycloaddition reaction of activated aziridines and palladium trimethylenemethane (Pd-TMM) complexes. The cycloaddition reaction generally proceeds enantiospecifically with ring opening at the least hindered site of the aziridine. Therefore, readily available enantiomerically pure 2-substituted aziridines can be utilized to prepare enantiomerically pure 2-substituted piperidines in good to excellent yield. The N-substituent on the aziridine proved to be crucial to the success of this reaction with only 4-toluenesulfonyl (Ts) and 4-methoxybenzenesulfonyl (PMBS) aziridines permitting smooth cycloaddition to take place. Additionally, spirocyclic aziridines have been found to participate in the [3+3] cycloaddition reaction, whereas 2,3-disubstituted aziridines can be applied to provide fused bicyclic piperidines, albeit in low yield. PMID- 12762729 TI - Highly enantiomerically enriched chlorophosphine boranes: synthesis and applications as P-chirogenic electrophilic blocks. AB - The stereoselective synthesis of P-chirogenic chlorophosphine boranes 4 was investigated by HCl acidolysis of the corresponding aminophosphine boranes 10. The reaction afforded the P-N bond cleavage with inversion of the configuration at the phosphorus center, leading to the chlorophosphine boranes 4 with high to excellent enantiomeric purities (80-99% ee), except in the case of the chloro-1 naphthylphenylphosphine borane 4d. Reaction conditions and workup significantly influence the enantiomeric purity of the product, with the exception of the o anisyl- and o-tolylchlorophenylphosphine boranes, 4b and 4c, which were found to be particularly stable even after purification by chromatography on silica gel. Reaction of the chlorophosphine boranes 4 with various nucleophiles, such as carbanions, phenolates, thiophenolates, or amides, afforded the corresponding organophosphorus borane complexes via P-C, P-O, P-S, and P-N bond formation, respectively, in 34-93% yield and with up to 99% ee. This work demonstrates the importance of chlorophosphine boranes 4 as new and powerful electrophilic building blocks for the highly stereoselective synthesis of P-chirogenic organophosphorus compounds. PMID- 12762730 TI - Palladium-catalyzed Suzuki-Miyaura cross-coupling reactions of potassium aryl- and heteroaryltrifluoroborates. AB - An extended study of the reactivity of potassium aryl- and heteroaryltrifluoroborates in Suzuki-Miyaura cross-coupling reactions is presented. The coupling of aryl- and electron-rich heteroaryltrifluoroborates with aryl and activated heteroaryl bromides proceeds readily under ligandless conditions. When deactivated aryl- and heteroaryltrifluoroborates are coupled with aryl and heteroaryl bromides and chlorides, a low loading (0.5-2%) of PdCl(2)(dppf).CH(2)Cl(2) efficiently catalyzes the reactions. Under either condition, reactions can generally be carried out in an open atmosphere. PMID- 12762731 TI - Influence of diene substitution on Diels-Alder reactions between vinyl dihydronaphthalenes and (SS)-2-(p-tolylsulfinyl)-1,4-benzoquinone. AB - The asymmetric Diels-Alder reaction between 2-(E-2-acetoxyvinyl)-8-tert-butyl-3,4 dihydronaphthalene (8) and enantiopure (SS)-2-(p-tolylsulfinyl)-1,4-benzoquinone (1) takes place exclusively on the unsubstituted C(5)-C(6) double bond of (SS)-1 with a very high control of the chemo-, regio-, and diastereoselectivity of the process affording tetracyclic sulfinyl derivative 13a possessing five stereogenic centers. The analogue diene 9, lacking the tert-butyl group, gave a less chemoselective reaction (C(2)-C(3)/C(5)-C(6): 60/40) in favor of reaction through the sulfoxide-substituted double bond C(2)-C(3) of 1. Steric effects of the remote tert-butyl group and electronic factors due to the OAc substituent are controlling the process. PMID- 12762732 TI - Evaluation of chiral oxazolines for the highly enantioselective diethylzinc addition to N-(diphenylphosphinoyl) imines. AB - On the basis of the principle that the incorporation of the structurally rigid and conformationally restricted skeleton in beta-amino alcohols is beneficial to the enantioselective diethylzinc addition to imines, a series of chiral oxazolines, which had been designed and conveniently prepared from commercially available (1S,2S)-2-amino-1-phenylpropane-1,3-diol, were applied in the diethylzinc addition to diphenylphosphinoyl imines to give high yields of 68-84% and excellent ee values of 90-95%. The configuration of the product was controlled by the chirality of the carbon bonded to the hydroxyl group in the oxazoline. Oxazolines bearing a para- or meta-substituted phenyl group generally offered higher enantioselectivity than those containing an ortho-substituted phenyl. The X-ray structures of 4f and 4j, in combination with the proposed transition state, preliminarily explained why oxazolines with a para- or meta substituent on the phenyl group gave higher enantioselectivities than those bearing an ortho-substituent. This successful example using chiral oxazolines to promote the titled reaction implies that a large family of chiral compounds containing an oxazoline ring moiety have the potential to be developed for promoting the highly enantioselective dialkylzinc addition to N (diphenylphosphinoyl) imines. PMID- 12762734 TI - Novel use of N-benzoyl-N,O-acetals as N-acylimine equivalents in asymmetric heterocycloaddition: an extended enantioselective pathway to beta-benzamido aldehydes. AB - For the first time, easily available N-(alpha-methoxyalkyl)amides were successfully used as synthetic equivalents of N-acylimines in an asymmetric heterocycloaddition process. The facial-controlled formation of 6 alkoxydihydrooxazines was thus achieved by SnCl(4)-promoted heterocycloaddition of (R)-O-vinyl pantolactone. By simple acidic hydrolysis of the crude heteroadducts, new beta-aryl- and beta-alkyl-substituted benzamido aldehydes were thus obtained in good overall yields with high enantioselectivities. PMID- 12762733 TI - Mechanism for basic hydrolysis of N-nitrosoguanidines in aqueous solution. AB - A kinetic study was carried out on the hydrolysis of two N-nitrosoguanidines, 1 nitroso-1-methyl-3-tolylsulfonylguanidine (TSGNO) and 1-nitroso-1-methyl-3 benzoylguanidine (BCGNO). We observed an absence of buffer catalysis using H(2)PO(4)(-)/HPO(4)(2)(-), H(3)BO(3)/H(2)BO(3)(-), and HCO(3)(-)/CO(3)(2)(-) regulators and a complex dependency of the rate constant on the pH. We discovered the existence of three simultaneous reaction paths: spontaneous decomposition of the neutral form of the N-nitrosoguanidine, decomposition of the monoanion, and decomposition through the form of the dianion. The analysis of the kinetic data has allowed us to obtain the acidity constant for the formation of the monoanion of the N-nitrosoguanidine, with values of p = 11.5. The reaction rate for the process through the monoanion, k(2), decreases as the acidity increases. The application of the principle of nonperfect synchronization shows that the basicity and reactivity do not correlate when there exists a possibility of stabilization of the negative charge by resonance. This behavior is consistent with the mechanism E1cB whereby the stabler the negative charge, the slower the elimination reaction. When dealing with the case of the elimination through the neutral form we observe that the reaction rate increases together with the capacity of stabilization of the positive charge on the nitrogen atom adjacent to the imino group. For the reaction through the dianion we used a maximum value of k(3) = 10(10) s(-)(1) to estimate the value of p for the formation of the dianion of the N-nitrosoguanidine, obtaining values of p < 24. PMID- 12762735 TI - Dienophilicity of imidazole in inverse electron demand Diels-Alder reactions. 4. Intermolecular reactions with 1,2,4-triazines. AB - Intermolecular inverse electron demand cycloadditions of 2-substituted imidazoles with various 1,2,4-triazines produced both imidazo[4,5-c]pyridines (3 deazapurines) and pyrido[3,2-d]pyrimid-4-ones (8-deazapteridines). The product distribution was controlled by reactant substituents and influenced by reaction temperature. A regioselective method for the preparation of 6-unsubstituted 1,2,4 triazines was also developed. By using this route to 8-deazapteridines, a new 8 deazafolate analogue was prepared. PMID- 12762736 TI - Lewis acid mediated reactions of zirconacyclopentenes with aldehydes affording homoallyl ketones via oppenauer-type oxidation. AB - Three different components involving alkynes, ethylene, and aldehydes were selectively integrated in a one-pot procedure to afford homoallyl ketones in good yields, via an effective combination of zirconocene-mediated C-C bond forming reactions and Lewis acid mediated organic transformation. Mechanistic studies revealed that a formal Oppenauer oxidation of seven-membered oxazirconacycles, generated in situ from the reactions of zirconacyclopentenes and aldehydes, was promoted by Lewis acid-aldehyde adducts. As a whole, the first aldehyde was incorporated into the product and the second aldehyde was reduced to an alcohol. Multiply deuterated homoallyl ketones could be readily prepared in high yields with more than 98% deuterium incorporation by using this method. PMID- 12762737 TI - General patterns in the photochemistry of pregna-1,4-dien-3,20-diones. AB - The photochemistry of six pregna-1,4-dien-3,20-diones has been compared and found to involve both the cyclohexadienone moiety in ring A and the isolated ketone at C-20. The two reactions take place proportionally to the fraction of light absorbed by each chromophore. The cross-conjugated ketone absorbs predominantly or exclusively at both 254 and 366 nm and undergoes the "lumi" rearrangement to bicyclo[3.1.0]hex-3-en-2-one. The quantum yield of the reaction diminished somewhat with increasing lambda(exc), e.g., for prednisolone Phi(254) (nm) = 0.42, Phi(366) (nm) = 0.3. A much stronger lowering is caused by halogen substitution in position 9 (by a factor of 3 for F, >50 for Cl), apparently due to a shortened triplet lifetime caused by heavy atom effect. At 310 nm, both chromophores absorb to a comparable degree and both may react. The reaction at C(20) ketone involves either quite efficient alpha-cleavage (C(17)-C(20)) for compounds bearing an acetal or hydroxyl function at C(17) or less effective (by a factor of ca. 10) hydrogen abstraction from the 18-methyl group in the other cases (finally resulting in Norrish II fragmentation or Yang cyclization). The results allow generalizing how the substitution pattern surrounding each chromophore affects the photoreactivity at that site and the competition between the two modes, allowing predicting the photochemistry of this family of antiinflammatory drugs. PMID- 12762738 TI - Easy access to novel substituted 6-aminoimidazo[1,2-a]pyridines using palladium- and copper-catalyzed aminations. AB - Convenient and efficient methods for the preparation of novel 6-aminoimidazo[1,2 a]pyridine derivatives are reported that utilized palladium- or copper-catalyzed methodology. The crystal structure for 6-N-methylanilinoimidazo[1,2-a]pyridine 10 is also described. PMID- 12762740 TI - Theoretical studies on cycloaddition reactions between the 2-aza-1,3-butadiene cation and olefins. AB - Density functional (B3LYP) calculations, using the 6-31G basis set, have been employed to study the title reactions. For the model reaction (H(2)C=C NH(+)=CH(2) + H(2)C=CH(2)), a complex has been formed with 6.2 kcal/mol of stabilization energy and the transition state is 4.0 kcal/mol above this complex, but 2.1 kcal/mol below the reactants. However, the substituent effects are quite remarkable. When ethene is substituted by electron-withdrawing group CN, the reaction could also yield six-membered-ring products, but the energy barriers are all more than 7 kcal/mol, which shows that CN group unfavors the reaction. The other substituents, such as CH(3)O and CH(3) groups, have also been considered in the present work, and the results show that they are favorable for the formation of six-membered-ring adducts. The calculated results have been rationalized with frontier orbital interaction and topological analysis. PMID- 12762739 TI - A flexible approach toward trisubstituted piperidines and indolizidines: synthesis of 6-epi-indolizidine 223A. AB - 2,5,6-Trisubstituted piperidines are readily prepared by a combination of an aza Achmatowicz oxidation of a furyl-substituted benzenesulfonamide followed by a conjugate addition to the resulting 2H-pyridone and subsequent addition of various nucleophiles to a transient N-sulfonyliminium ion. The stereochemistry of the conjugate addition product is the result of axial attack from the face opposite the diaxial substituents at C(2) and C(6). This can be attributed to steric hindrance between the pseudoaxially oriented 2,6-substituents and the equatorially approaching nucleophile, thereby leading to the exclusive formation of the kinetically favored axial 1,4-adduct. Indolizidine alkaloid 223A was isolated from a skin extract of a Panamanian population of the dendrobatid Dendrobates pumilio Schmidt (Dendrobatidae). Synthesis of the originally proposed structure of this alkaloid was achieved in 13 steps in 13.1% overall yield by using an aza-Achmatowicz oxidative rearrangement and a diastereoselective 1,4 conjugate addition as the key steps. The structure of the natural 223A alkaloid (5b) differs from that of the epi-isomer 5a synthesized in this study in the configuration at the 6-position of the indolizidine ring. PMID- 12762741 TI - Alkylation of nitroaromatics with trialkyborane. AB - When p-dinitrobenzene is reacted with Et(3)B in t-BuOH or THF in the presence of t-BuOK, it yields p-nitroethylbenzene. In this report we examine the scope of this transformation by monitoring the effect of various parameters on the reaction. It has been found that the reaction is extremely sensitive to temperature and rather insensitive to the base-solvent combination used. It is also insensitive to the steric hindrance of the base: good yields were obtained using sodium 2,6-diisopropylphenoxide or when using NaH. Alkylation was obtained with a large variety of alkylboranes ranging from linear to polycyclic. Yields drop significantly if one of the nitro groups is replaced by another electron withdrawing group. In all cases studied (CHO, PHCO, SO(2)Ph, and CN), it is the latter group which was preferentially displaced by the alkyl group. According to the suggested mechanism, the radical anion of the substrate combines with the alkyl radical released from the boranyl radical to form a Meisenheimer complex. The reaction takes place at the ring carbon bearing the highest spin density in accordance with ab initio calculations at the B3LYP/6-31+G level. PMID- 12762742 TI - Investigation of a stereoselective co-mediated rearrangement reaction. AB - A stereocontrolled approach to alpha-alkyl beta-alkynyl cyclohexanones is reported through a Lewis acid mediated rearrangement reaction of enol ethers bearing an Co-alkyne moiety. The reaction proceeds with high levels of stereoselectivity in the presence of Ti- and B-Lewis acids to provide a range of alpha,beta-disubstituted cyclohexanones in high yield although the products are prone to epimerization at the alpha-position in the presence of the B-promoter system. The potential for an enantioselective variant of this process is outlined, and a rationale for the observed stereochemical trends and detailed structural analyses of the ketone products are described. PMID- 12762743 TI - Asymmetric total synthesis of (-)-alkaloid 223A and its 6-epimer. AB - The enantiopure gamma-amino alcohols 7 and 18 are prepared by using the diastereoselective Michael addition of lithium N-benzyl (R)-alpha methylbenzylamide to alpha,beta-unsaturated esters as a key step. The Michael addition of 7 or 18 to an alkynone 8 followed by an intramolecular cyclization afford the cyclic enamine 10 or 20, which are subjected to the diastereoselective hydrogenation, and the subsequent transformations provide 6-epi-alkaloid 223A and alkaloid 223A, respectively. PMID- 12762745 TI - Preparation of highly fluorinated cyclopropanes and ring-opening reactions with halogens. AB - Various highly fluorinated cyclopropanes 1 were prepared by reaction of the appropriate fluorinated olefins with hexafluoropropylene oxide (HFPO) at 180 degrees C. The fluorinated nitrile 1e was converted to the triazine derivatives 2a and 2b by catalysis with Ag(2)O and NH(3)/(CF(3)CO)(2)O, respectively. The fluorinated cyclopropanes reacted with halogens at elevated temperatures to provide the first useful, general synthesis of 1,3-dihalopolyfluoropropanes. At 150-240 degrees C, hexafluorocyclopropane and halogens X(2) produce XCF(2)CF(2)CF(2)X (X = Cl, Br, I) in 50-80% isolated yields. Pentafluorocyclopropanes c-C(3)F(5)Y [Y = Cl, OCF(3), OC(3)F(7) and OCF(2)CF(CF(3))OCF(2)CF(2)Z; Z = SO(2)F, CN, CO(2)Me] react regiospecifically at 150 degrees C to give XCF(2)CF(2)CFXY, c-C(3)F(5)Br reacts regioselectively with Br(2) to give a 16.7:1 mixture of BrCF(2)CF(2)CFBr(2):BrCF(2)CFBrCF(2)Br, whereas c-C(3)F(5)H reacts unselectively with I(2) to produce a statistical 2:1 mixture of ICF(2)CF(2)CFHI:ICF(2)CFHCF(2)I. Tri- and di(pentafluorocyclopropyl) derivatives 2 also undergo ring-opening reaction with halogens to give 16 and 17. Upon treatment of tetrafluorocyclopropanes 1j, 1k, and 1l with Br(2) or I(2), ring opening occurred exclusively at substituted carbons to give XCF(2)CF(2)CXY(2). Thermolysis of the ring-opened product ICF(2)CF(2)CFIOR(F) at 240 degrees C gave R(F)I and ICF(2)CF(2)COF in high yields. PMID- 12762744 TI - Stereoselective synthesis of tetrasubstituted 2,3-dihydrofurans by one-step cyclization of beta-ketosulfides of benzothiazole and aldehydes in ionic liquids. AB - A stereoselective synthesis of tetrasubstituted 2,3-dihydrofurans was carried out in n-butylpyridinium tetrafluoroborate ([bpy(+)][BF(4)(-)]) as solvent. The reaction proceeds smoothly in one step starting from simple materials such as aldehydes and beta-ketosulfides of benzothiazole. A comparison between several ionic liquids (ILs) is presented, and the role of the benzothiazolyl moiety is discussed. Workup proved to be very easy and recycling of IL possible. PMID- 12762746 TI - Efficient and rapid synthesis of oligo(p-phenylenevinylene) via iterative coherent approach. AB - A general and controlled bidirectional growth strategy enables a very rapid and efficient construction of OPV compounds possessing functional groups such as aldehyde and mercapto groups at both ends. The strategy employs only one reaction type with high yields and stereoselectivities to grow the conjugated chains, eliminating the need for protecting groups in the overall intermediates. PMID- 12762747 TI - Synthesis of phytuberin. 4-endo-tet acid-catalyzed cyclization of alpha-hydroxy epoxides. AB - The total synthesis of phytuberin, a phytoalexin of the Solanum genus, from (-) alpha-santonin is reported. The key steps include (a) reductive cleavage of the C O bond of the gamma-lactone with concomitant protection of the C1 double bond, (b) Sharpless stereocontrolled hydroxy-assisted epoxidation of allylic alcohol 6 and simultaneous deprotection of the C1 double bond, (c) a rare 4-endo-tet acid catalyzed cyclization of an alpha-hydroxy epoxide, and (d) an unprecedented 4-exo selenocyclization of a homoallylic alcohol. PMID- 12762748 TI - Facile and efficient synthesis of meso-arylamino- and alkylamino-substituted porphyrins via palladium-catalyzed amination. AB - meso-Arylamino- and alkylamino-substituted porphyrins were efficiently synthesized by reactions of meso-halogenated porphyrins with amines via palladium catalyzed amination. The combination of palladium acetate and the commercially available phosphine ligand bis(2-diphenylphosphinophenyl) ether (DPEphos) is effective for catalyzing the couplings of both [5-bromo-10,20-diphenyl porphyrino]zinc(II) and [5,15-dibromo-10,20-diphenylporphyrino]zinc(II) with amines to give the corresponding monoamino- and diamino-substituted porphyrins in high yields under mild conditions. The corresponding halogenated free-base porphyrins also underwent the cross-coupling reactions efficiently under similar catalytic conditions. PMID- 12762749 TI - N7-guanine as a C+ mimic in hairpin aeg/aepPNA-DNA triplex: probing binding selectivity by UV-Tm and kinetics by fluorescence-based strand-invasion assay. AB - N7-substituted guanine (N7G) has been introduced into aminoethylglycyl bisPNA (7) as a C(+) mimic to achieve pH-independent triplex formation with complementary DNA sequences. The introduction of chiral, cationic aminoethylprolyl units with C(+) and C(+) mimic N7G in the backbone of bisPNAs (8, 9) influenced the recognition of complementary DNA in an orientation-selective manner. A simple fluorescence assay is developed to examine the process of strand invasion of target DNA duplex by these modified bisPNAs and comparative results of the study employing triplex forming polypyrimidine (C/T) (6, 8) and purine-pyrimidine (N7G/T) mixmer-bisPNAs (7, 9) are presented. PMID- 12762750 TI - Iterative synthesis of Leishmania phosphoglycans by solution, solid-phase, and polycondensation approaches without involving any glycosylation. AB - A general strategy (solution, solid-phase, and polycondensation) for the synthesis of antigenic phosphoglycans (PG) of the protozoan parasite Leishmania is presented. Phosphoglycans constitute the variable structural and functional domain of major cell-surface lipophosphoglycan (LPG) and secreted proteophosphoglycan (PPG), the molecules involved in infectivity and survival of the Leishmania parasite inside human macrophages. We have shown that the chemically labile, anomerically phosphodiester-linked phosphoglycan repeats can be assembled in an iterative and efficient manner from a single key intermediate, without involving any glycosylation steps. Furthermore, the phosphoglycan chain can be extended toward either the nonreducing (6'-OH) or the reducing (1-OH) end. We also describe a new and efficient solid-phase methodology to construct phosphoglycans based on design and application of a novel cis-allylphosphoryl solid-phase linker that enabled the selective cleavage of the first anomeric phosphodiester linkage without affecting any of the other internal anomeric phosphodiester groups of the growing PG chain on the solid support. The strategy to construct larger phosphoglycans in a one-pot synthesis by polycondensation of a single key intermediate is also described, enabling CD spectrometric measurements to show the helical nature of phosphoglycans. Our versatile synthetic approach provides easy access to Leishmania phosphoglycans and the opportunity to address key immunological, biochemical, and biophysical questions pertaining to the phosphoglycan family (LPG and PPG) unique to the parasite. PMID- 12762751 TI - Preparation of tri- and difluoromethylsilanes via an unusual magnesium metal mediated reductive tri- and difluoromethylation of chlorosilanes using tri- and difluoromethyl sulfides, sulfoxides, and sulfones. AB - A new and efficient method for the preparation of tri- and difluoromethylsilanes using magnesium metal-mediated reductive tri- and difluoromethylation of chlorosilanes is reported using tri- and difluoromethyl sulfides, sulfoxides, and sulfones. The byproduct of the process is diphenyl disulfide. Since phenyl trifluoromethyl sulfone, sulfoxide, and sulfide are readily prepared from trifluoromethane (CF(3)H) and diphenyl disulfide, the method can be considered to be catalytic in diphenyl disulfide for the preparation of (trifluoromethyl)trimethylsilane (TMS-CF(3)) from non-ozone-depleting trifluoromethane. PMID- 12762752 TI - A convergent solution-phase synthesis of the macrocycle Ac-Phe-[Orn-Pro-D-Cha-Trp Arg], a potent new antiinflammatory drug. AB - Relatively few cyclic peptides have reached the pharmaceutical marketplace during the past decade, most produced through fermentation rather than made synthetically. Generally, this class of compounds is synthesized for research purposes on milligram scales by solid-phase methods, but if the potential of macrocyclic peptidomimetics is to be realized, low-cost larger scale solution phase syntheses need to be devised and optimized to provide sufficient quantities for preclinical, clinical, and commercial uses. Here, we describe a cheap, medium scale, solution-phase synthesis of the first reported highly potent, selective, and orally active antagonist of the human C5a receptor. This compound, Ac-Phe[Orn Pro-d-Cha-Trp-Arg], known as 3D53, is a macrocyclic peptidomimetic of the human plasma protein C5a and displays excellent antiinflammatory activity in numerous animal models of human disease. In a convergent approach, two tripeptide fragments Ac-Phe-Orn(Boc)-Pro-OH and H-d-Cha-Trp(For)-Arg-OEt were first prepared by high-yielding solution-phase couplings using a mixed anhydride method before coupling them to give a linear hexapeptide which, after deprotection, was obtained in 38% overall yield from the commercially available amino acids. Cyclization in solution using BOP reagent gave the antagonist in 33% yield (13% overall) after HPLC purification. Significant features of the synthesis were that the Arg side chain was left unprotected throughout, the component Boc-d-Cha-OH was obtained very efficiently via hydrogenation of d-Phe with PtO(2) in TFA/water, the tripeptides were coupled at the Pro-Cha junction to minimize racemization via the oxazolone pathway, and the entire synthesis was carried out without purification of any intermediates. The target cyclic product was purified (>97%) by reversed-phase HPLC. This convergent synthesis with minimal use of protecting groups allowed batches of 50-100 g to be prepared efficiently in high yield using standard laboratory equipment. This type of procedure should be useful for making even larger quantities of this and other macrocyclic peptidomimetic drugs. PMID- 12762753 TI - Synthesis of functionalized 1,4-cyclohexadienes through intramolecular anionic dearomatization of N-alkyl-N-benzyldiphenylphosphinamides. Insight into the reaction mechanism. AB - A generalization of the intramolecular nucleophilic dearomatization-electrophilic alkylation reactions of N-alkyl-N-benzyldiphenylphosphinamide anions is presented. The process has been optimized by analyzing the effects of metalation and quench times, additives, the nature of the electrophiles used (MeI, CF(3)SO(3)Me, Me(3)O(+)BF(4)(-), AllylBr, PhCH(2)Br, BrCH(2)CO(2)Me, and RCH=O, where R = Ph, 4-Cl-C(6)H(4), 4-MeO-C(6)H(4), and (i)()Pr), and the alkyl substituent linked to the nitrogen of the phosphinamide. Both HMPA and DMPU act as catalysts. The latter proved to be much more efficient for obtaining high yields of substituted tetrahydrobenzo[c][1,2]-1lambda(5)-phospholes containing a 1,4-cyclohexadiene system with very high regio- and diastereoselectivity. Steric effects in the neighborhood of the benzylic anion tend to decrease the stereoselectivity of the anionic cyclization. The optimization study also served to shed light on the reaction mechanism of the dearomatization process by identifying several intermediate species and showing the reversibility of the anionic cyclization step as well as of the reaction with aldehydes. PMID- 12762754 TI - Synthesis of 2,6-bridged piperazine-3-ones by N-acyliminium ion chemistry. AB - Several 2-substituted and 2,5-disubstituted piperazine-3,6-diones were synthesized starting from readily available alpha-amino acids. After activation of a lactam carbonyl via introduction of a methoxycarbonyl group onto nitrogen, this carbonyl was selectively reduced. Treatment of the resulting urethane with protic acid generated the corresponding N-acyliminium ion, which was trapped by a nucleophilic C2-side chain to provide 2,6-bridged piperazine-3-ones. Several aromatic, heteroaromatic, and nonaromatic side chains were used as pi nucleophiles. In addition, the effect of the presence of a C5-methyl group on the stereochemical outcome of the cyclization was examined. PMID- 12762755 TI - Hydroboration of C(100) surface, fullerene, and the sidewalls of single-wall carbon nanotubes with borane. AB - Hydroboration of three allotropes of carbon, i.e., diamond (100) surface, [60]fullerene, and single-wall carbon nanotubes (SWNTs), with borane (BH(3)) has been explored by means of quantum chemical calculations. The calculations predicted that the hydroboration of C(60) and the C(100)-2x1 surface occurs readily, whereas the hydroboration of the sidewall of an armchair (5,5) SWNT is thermoneutral with a barrier height of 11.5 kcal/mol. This suggests that sidewall hydroboration, if viable, would be highly reversible on the (5,5) SWNT. The as hydroborated carbonous materials can be good starting points for further chemical modification and manipulation of these carbonous materials, given the abundant chemistry of organoboranes. PMID- 12762756 TI - Stereocontrolled synthesis of ara-type cyclohexenyl nucleosides. AB - A highly stereocontrolled synthesis of a new class of carbocyclic nucleosides, ara-type cyclohexenyl nucleosides, was developed. The key intermediate (+/-)-9 was obtained after a series of transformations starting from easily available endo-bicyclo carboxylic acid (+/-)-3. The allylic hydroxyl group of (+/-)-9 was masked via oxidation with manganese dioxide and released, after protection of the 2'-hydroxyl group, via reduction with NaBH(4) in the presence of CeCl(3).7H(2)O. The base moiety was introduced with use of the Mitsunobu methodology. PMID- 12762757 TI - Understanding the unusual regioselectivity in the nucleophilic ring-opening reactions of gem-disubstituted cyclic sulfates. Experimental and theoretical studies. AB - The regioselectivity of the nucleophilic ring-opening reactions of three gem disubstituted cyclic sulfates with sodium azide has been studied from both experimental and theoretical viewpoints. It is found that, depending on the substituent present in the cyclic sulfate, the reaction displays reversed regioselectivity, which allows one or another regioisomer to be obtained with selectivities greater than 4:1. The theoretical calculations show that, contrary to previous understanding, the intrinsic preference in all cases is azide attack at the less-substituted C(beta) position, a consequence of similar stereoelectronic effects in the three sulfates considered. The observed preference for C(alpha) attack in the case of the ester sulfate is explained in terms of differential solvent effects, which are in turn due to subtle differences in the charge transfer in the different transition structures. PMID- 12762758 TI - Highly enantioselective synthesis of alpha-alkyl-alanines via the catalytic phase transfer alkylation of 2-naphthyl aldimine tert-butyl ester by using O(9)-allyl N(1)-2',3',4'-trifluorobenzylhydrocinchonidinium bromide. AB - Systematic investigations to develop an efficient enantioselective synthetic method for alpha-alkyl-alanine by catalytic phase-transfer alkylation were performed. The alkylation of 2-naphthyl aldimine tert-butyl ester, 1E, with RbOH and O(9)-allyl-N-2',3',4'-trifluorobenzylhydrocinchonidinium bromide, 6, at -35 degrees C showed the highest enantioselectivities, up to 96% ee. PMID- 12762759 TI - A direct synthesis of O-methyl claussequinone. AB - The reaction of a chromene with BH(3) followed by treatment with benzoquinone and air is the key step in a direct entry to O-methyl claussequinone. PMID- 12762761 TI - Concise enantioselective syntheses of quinolactacins A and B through alternative Winterfeldt oxidation. AB - Enantioselective total syntheses of (+)-quinolactacin B and (+)-quinolactacin A2 through asymmetric Pictet-Spengler cyclization and KO(2) oxidation-an alternative Winterfeldt condition-are described. PMID- 12762760 TI - Straightforward synthesis of panaxytriol: an active component of Red Ginseng. AB - A total synthesis of (3R,9R,10R)-panaxytriol (1) was accomplished enantioselectively (40% overall yield; 30% for the longest sequence). A key step was a Cadiot-Chodkiewicz cross-coupling reaction on two fragments containing, in the aggregate, three unprotected hydroxyl groups. One fragment was synthesized by a highly enantioselective reduction of an enynone. The other arose from a highly enantioselective dihydroxylation of an allylic alcohol. PMID- 12762762 TI - Identification of regioisomers in a series of N-substituted pyridin-4-yl imidazole derivatives by regiospecific synthesis, GC/MS, and 1H NMR. AB - The regiospecific synthesis of 2a (Scheme 3), a novel and potent pyridinyl imidazole inhibitor of p38 MAP (mitogen-activated protein) kinase, and the regioselective preparation of its regioisomer 2b (Scheme 4) are described. Chromatographic and spectroscopic data are presented, which in this class of compounds allow the unambiguous identification of regioisomers prepared by a nonregiospecific synthetic strategy. Biological data demonstrating the importance of the correct regiochemistry for inhibition of p38 are given. PMID- 12762763 TI - Synthesis of griseolic acid analogues: regioselective alpha-facial [1,2] migration in the rhodium acetate catalyzed reaction of D-glucose derived alpha diazo-beta-keto ester. AB - This paper describes an efficient route for the synthesis of known and novel griseolic acid analogues 1d and 1e, respectively. The key intermediate dioxabicyclo derivative 6, with the required stereochemical orientation at C6, was obtained by rhodium acetate catalyzed reaction of d-glucose derived alpha diazo-beta-keto ester 5 in a novel high-yielding methodology. PMID- 12762765 TI - Highly efficient rhodium/monodentate phosphoramidite catalyst and its application in the enantioselective hydrogenation of enamides and alpha-dehydroamino acid derivatives. AB - An easily prepared and highly efficient monodentate phosphoramidite ligand derived from BINOL, (S)-2,2'-O,O-(1,1'-binaphthyl)-dioxo-N,N diethylphospholidine, was examined in the hydrogenation of both enamides and alpha-dehydroamino acid derivatives. The catalyst provided remarkably high enantioselectivities (up to 99.6% ee for enamides and >99.9% ee for alpha dehydroamino acid derivatives). PMID- 12762764 TI - Zinc carbenoid-mediated chain extension: preparation of alpha,beta-unsaturated gamma-keto esters and amides. AB - An efficient one-pot preparation of alpha,beta-unsaturated-gamma-keto esters and amides has been developed. A zinc carbenoid-mediated chain extension of a beta dicarbonyl substrate provides access to an intermediate zinc enolate, which is treated sequentially with a halogen and amine base. This method has been applied to a variety of ester and amide starting materials, as well as to amino acid derived substrates and to a formal synthesis of (R,R)-(-)-pyrenophorin. PMID- 12762767 TI - Simple diastereoselectivity on addition of alpha-haloalkyl Grignard reagents to benzaldehyde. AB - The addition of alpha-haloalkyl Grignard reagents to benzaldehyde occurs with simple diastereoselectivity substantially higher than that of the corresponding lithium reagents. Reaction in the presence of dimethyl-aluminum chloride suppresses subsequent Oppenauer oxidation of the resulting Mg-alkoxides by excess benzaldehyde. PMID- 12762766 TI - Readily prepared chiral P,N ligands and their applications in cu-catalyzed enantioselective conjugate additions. AB - A new type of phosphite-pyridine (P,N) ligand derived from (S)-NOBIN and (S) BINOL was employed in Cu(I)-catalyzed conjugate addition of diethylzinc to chalcones. The new P,N ligands were highly efficient in the copper-catalyzed enantioselective 1,4-conjugate additions of diethylzinc to acyclic enones, and up to 97% ee was achieved. PMID- 12762768 TI - Sterically crowded bicyclo[1.1.0]butane radical cations. AB - The variability of carbon-carbon single bonds by steric and electronic effects is probed by DFT calculations of sterically crowded bicyclo[1.1.0]butanes and their radical cations. The interplay of sterics and electronics on the gradual weakening and breaking of bonds was studied by investigating bridgehead substitution in 1,3-di-tert-butylbicyclo[1.1.0]butane and 2,2',4,4'-tetramethyl 1,3-di-tert-butylbicyclo[1.1.0]butane and geminal substitution in 2,2'-di-tert butylbicyclo[1.1.0]butane and 2,2',4,4'-tetra-tert-butylbicyclo[1.1.0]butane. Bridgehead substitution leads to a lengthening of the central bond, whereas bisubstitution on the geminal carbon leads to a shortening of this bond due to a Thorpe-Ingold effect. Although the character of the central bond can be modulated by substitution and electron transfer over a range of 0.35 A, the state forbidden ring planarization does not occur. Sterically crowded bicyclo[1.1.0]butane radical cations are therefore promising candidates for the investigation of extremely long carbon-carbon single bonds. PMID- 12762769 TI - Wet silica-supported permanganate for the cleavage of semicarbazones and phenylhydrazones under solvent-free conditions. AB - Wet silica-supported potassium permanganate was used as an inexpensive and efficient reagent for conversion of semicarbazones and phenylhydrazones to the corresponding carbonyl compounds under solid-state conditions. PMID- 12762770 TI - A novel and expeditious approach to unusual spirolactam building blocks. AB - A rapid access to 7-azaspiro[4.5]decan-6-ones 1 involving three regio- and chemoselective reactions starting from tetrabromonorbornyl derivatives is described. The alkaline H(2)O(2) cleavage reaction of monosubstituted alpha diketones 9 furnished the potential bridged bicyclic lactones 10in a highly regio and stereoselective manner. The radical-mediated, intermolecular bridgehead C-C bond formation of the versatile bridged lactones 10 with acrylonitrile followed by LAH reduction of the adduct 13 intriguingly leads to the formation of novel spirolactam building blocks 1. PMID- 12762771 TI - Mechanistic investigation of substrate oxidation by Ce(IV) reagents in acetonitrile. AB - Rates and activation parameters for the Ce(4+)-mediated oxidation of a beta-keto ester, a beta-diketone, and a beta-keto silyl enol ether were determined in acetonitrile. In the case of the dicarbonyls, the enol content of the substrate impacts the rate of oxidation by Ce(4+), predominantly through contributions from DeltaH(). For the silyl enol ether, the transition state for oxidation by Ce(4+) is substantially more ordered than it is for the beta-keto ester or the beta diketone. PMID- 12762772 TI - Himeradine A, a novel C27N3-type alkaloid from Lycopodium chinense. AB - A novel C(27)N(3)-type Lycopodium alkaloid consisting of a fastigiatine-type skeleton (C(16)N(2)) and a quinolizidine moiety (C(11)N), himeradine A (1), has been isolated from the club moss Lycopodium chinense, and the structure and relative stereochemistry were elucidated on the basis of spectroscopic data. PMID- 12762773 TI - An efficient route to 3-aryl-substituted quinolin-2-one and 1,8-naphthyridin-2 one derivatives of pharmaceutical interest. AB - Reaction of arylacetic ester enolates with 2-alkoxy-4H-3,1-benzoxazin-4-ones offers a short and versatile synthetic route to 3-aryl-4-hydroxyquinolin-2(1H) ones, through the cyclization of the beta-ketoesters produced. Similar reactions of 4H-pyrido[2,3-d][1,3]oxazin-4-ones with ester enolates afford 1-acyl-4-hydroxy 1,8-naphthyridin-2(1H)-ones in a convenient two-step, one-pot procedure. PMID- 12762774 TI - Nitrile biotransformations for highly efficient and enantioselective syntheses of electrophilic oxiranecarboxamides. AB - Catalyzed by a nitrile hydratase/amidase-containing microbial Rhodococcus sp. AJ270 whole-cell catalyst, a number of racemic trans-2,3-epoxy-3 arylpropanenitriles 1 underwent rapid and efficient hydrolysis under very mild conditions to afford 2R,3S-2-arylglycidamides 2 in excellent yield with enantiomeric excess higher than 99.5%. The overall enantioselectivity of the biotransformations originated from the combined effects of a dominantly high 2S enantioselective amidase and low 2S-enantioselective nitrile hydratase involved in the cell. The influence of the substrates on both reaction efficiency and enantioselectivity was also discussed in terms of steric and electronic effects. PMID- 12762775 TI - Zirconium(IV) chloride catalyzed new and efficient protocol for the selective cleavage of p-methoxybenzyl ethers. AB - A highly selective and efficient method for the unmasking of p-methoxybenzyl (PMB) ethers and esters has been developed by use of 20 mol % of zirconium(IV) chloride as Lewis acid in acetonitrile. The present method is very fast, and the conditions are tolerable to a variety of acid/base-sensitive protecting groups and substrates such as carbohydrates, terpenes, and amino acids. The products are obtained in good to high yields. PMID- 12762776 TI - New, improved procedure for the synthesis of structurally diverse N-spiro C2 symmetric chiral quaternary ammonium bromides. AB - Selective, direct ortho magnesiation of (S)-2,2'-bis(isopropoxycarbonyl)-1,1' binaphthyl (6) has been achieved under mild conditions, using magnesium bis(2,2,6,6-tetramethylpiperamide) [Mg(TMP)(2)]. In combination with the subsequent reaction with the appropriate electrophiles, bromine and iodine, this method constitutes a key step in establishing a new and concise synthetic route to a wide variety of N-spiro C(2)-symmetric chiral quaternary ammonium bromides of type 1. PMID- 12762777 TI - Optimization of the abnormal Beckmann rearrangement: application to steroid 17 oximes. AB - A novel and practical procedure was developed for the abnormal Beckmann rearrangement of steroid 17-oximes. Treatment of the 17-oximes with TFA/CH(OMe)(3) in boiling THF for 2 h gives the corresponding 13,17-seco alkene nitrile products in unprecedented high yields (70-92%). Since the alkene nitriles can be subsequently converted into 18-norsteroids, this general method provides a highly efficient route to these biologically important compounds and, by extension, to other structurally related natural products. PMID- 12762778 TI - Expedient one-pot synthesis of novel chiral 2-substituted 5-phenyl-1,4 benzodiazepine scaffolds from amino acid-derived amino nitriles. AB - An efficient and stereocontrolled synthesis of phenylalanine- and tryptophan derived 5-phenyl-1,4-benzodiazepines is described. This new methodology involves, as a key step, the synthesis of 5-phenyl-2,3-dihydro-1H-1,4-benzodiazepines by a one-pot cyano reduction and reductive cyclization of the appropriate amino nitrile, which were obtained via a modified Strecker reaction of N-protected alpha-amino aldehydes with 2-aminobenzophenone and trimethylsilyl cyanide. The subsequent reduction of these 2,3-dihydro-1H-1,4-benzodiazepines, followed by regioselective alkylation or acylation at position 4, led to 2,4-disubstituted-5 phenyl-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine. PMID- 12762779 TI - Cyclization of aryl acyl radicals generated from S-(4-cyano)phenyl thiolesters by a nickel complex catalyzed electroreduction. AB - Aromatic acyl radicals generated from S-(4-cyano)phenyl 2-alkenylthiobenzoate by a nickel complex catalyzed electroreduction undergo 5- and 6-exo cyclization to give 1-indanone and dihydro-1-naphthalenone derivatives, respectively. PMID- 12762780 TI - Copper-catalyzed synthesis of unsymmetrical triarylphosphines. AB - Various triarylphosphines have been prepared by coupling diphenylphosphine with aryl iodides with catalytic amounts of CuI in the presence of either K(2)CO(3) or Cs(2)CO(3), in good yields. This method can tolerate a variety of functional groups and does not require the use of expensive additives, or harsh reaction conditions, and is palladium free. PMID- 12762781 TI - The Michael addition of indoles to alpha,beta-unsaturated ketones catalyzed by CeCl3.7H2O-NaI combination supported on silica gel. AB - Alkylation of indoles by means of the Michael addition has been the subject of a number of investigation. It is well established that regioselectivity in the additions of indoles to electron-deficient alkenes is strongly controlled by the reaction medium. In a continuation of the work on developing greener and cleaner technologies, the cerium(III) chloride heptahydrate and sodium iodide combination supported on silica gel catalyzes the alkylation of various indoles with alpha,beta-unsaturated ketones giving 3-(3-oxoalkyl)indole derivatives in good yields. The substitution on the indole nucleus occurred exclusively at the 3 position, and N-alkylation products have not been observed. PMID- 12762782 TI - A convenient one-pot synthesis of 1,8-naphthyridones. AB - In this paper, we disclose an efficient one-pot procedure for the preparation of substituted 1,8-naphthyridin-4-one analogues. Previous efforts to effect this type of transformation were complicated by the formation of benzene tricarboxylate. Via the use of excess base, the impurity formation was completely inhibited. This allowed for the clean preparation of the desired intermediate and subsequent formation of naphthyridone analogues in a single flask, which could then be crystallized directly from the reaction mixture in good yield and high purity. PMID- 12762784 TI - Effects of alpha-ammonium, alpha-phosphonium, and alpha-sulfonium groups on C-H bond dissociation energies. AB - C-H bond dissociation energies of alpha-ammonium-, alpha-phosphonium-, or alpha sulfonium-substituted methanes and toluenes were calculated to a precision of 1-2 kcal/mol. It was found that alpha-ammonium, alpha-phosphonium, and alpha sulfonium groups all destabilize a methyl radical. alpha-Ammonium also destabilizes a benzyl radical, whereas alpha-phosphonium and alpha-sulfonium either slightly stabilize or destabilize a benzyl radical depending on their alkylation state. PMID- 12762783 TI - Scope of enantioselective palladium(II)-catalyzed aerobic alcohol oxidations with (-)-sparteine. AB - Evaluation of the substrate scope for Pd(II)/ (-)-sparteine catalyzed aerobic oxidative kinetic resolution of secondary alcohols is disclosed. An improved system is found with use of tert-butyl alcohol solvent in which benzylic and aliphatic alcohols as well as alcohols containing olefins are effectively oxidatively resolved. For substrates that successfully undergo oxidative kinetic resolution, k(rel) values are generally between 10 and 20. Successful scale-up of various substrates to 10-mmol scale is described. Extension to oxidative desymmetrization of 1,3-meso-diols is successful with enantiomeric excesses ranging from 78 to 85%. PMID- 12762787 TI - Potential cancer chemopreventive constituents of the seeds of Dipteryx odorata (tonka bean). AB - A new cassane diterpene, dipteryxic acid (1), and a new isoflavonolignan, 5 methoxyxanthocercin A (2), as well as four known active compounds, isoliquiritigenin (3), 6,4'-dihydroxy-3'-methoxyaurone (4), sulfuretin (5), and (+/-)-balanophonin (6), and five known inactive compounds, butin, eriodictyol, 7 hydroxychromone, 7,3'-dihydroxy-8,4'-dimethoxyisoflavone, and (-)-lariciresinol, were isolated from an ethyl acetate-soluble extract of the seeds of Dipteryx odorata, using a bioassay based on the induction of quinone reductase (QR) in cultured Hepa 1c1c7 mouse hepatoma cells to monitor chromatographic fractionation. The structures of compounds 1 and 2 were elucidated by spectroscopic data interpretation. Single-crystal X-ray diffraction analysis was used to confirm the relative stereochemistry of compound 1. Selected compounds (3 5) were evaluated in a mouse mammary organ culture assay, with isoliquiritigenin (3) found to exhibit 76% inhibition at a dose of 10 microg/mL. PMID- 12762788 TI - Synthetic approach to exo-endo cross-conjugated cyclohexadienones and its application to the syntheses of dehydrobrachylaenolide, isodehydrochamaecynone, and trans-isodehydrochamaecynone. AB - Methodology for synthesis of exo-endo cross-conjugated dienones with trans- and cis-decalin systems has been reported. Bromination of the silyl enol ether of alpha'-methyl alpha,beta-unsaturated ketones with PTAB and successive dehydrobromination of the resulting alpha'-bromo-alpha'-methyl alpha,beta unsaturated ketones under three conditions (DBU/PhH; TBAF/THF; Li(2)CO(3), LiBr/DMF) gave the desired exo-endo cross-conjugated dienones in good yield. This method was applied to the syntheses of dehydrobrachylaenolide (1), isodehydrochamaecynone (5c), and trans-isodehydrochamaecynone (11) starting from tuberiferine (7), chamaecynone (5a), and trans-chamaecynone (9). Eudesmanolides possessing an alpha-methylene gamma-lactone moiety, i.e., 1, 7, and 13, exhibited significant inhibitory activity toward the induction of the intercellular adhesion molecule-1 (ICAM-1). Compound 1 showed greater activity than 7 and 13. All compounds possessing an ethynyl group, 5d, 9, 11, and 14, showed the same degree of termiticidal activity, and the exo-endo cross-conjugated dienone structure in 11 had no influence on the activity. PMID- 12762789 TI - Continuing studies on the cyanobacterium Lyngbya sp.: isolation and structure determination of 15-norlyngbyapeptin A and lyngbyabellin D. AB - Re-collections of the cyanobacterium Lyngbya sp. have yielded two more members of the lyngbyapeptin and lyngbyabellin families. The gross structures of 15 norlyngbyapeptin A (1) and lyngbyabellin D (3) were deduced through standard 2D NMR techniques, with the absolute configuration of both elucidated through degradation and comparison with commercially available and synthetic standards. Degradation to the alpha-amino acid and NOE correlations determined the absolute and relative configuration of the 4-amino-3-hydroxy-5-methylhexanoic acid unit in 3. Lyngbyabellin D (3) displayed an IC(50) value of 0.1 microM against the KB cell line. PMID- 12762790 TI - Rhuschalcones II-VI, five new bichalcones from the root bark of Rhus pyroides. AB - Biflavonoids detected in trace amounts in an earlier investigation of the twigs of Rhus pyroides have now been found in the root bark of this species. These new flavonoids belong to a rare bichalcone class and have been identified as 2',4',4' ',2' ",4' "-pentahydroxy-4-O-5' "-bichalcone (rhuschalcone II, 2), 2',4',4' ',2' "-tetrahydroxy-4' "-methoxy-4-O-5' "-bichalcone (rhuschalcone III, 3), 4,2',4' ',2' "-tetrahydroxy-4' "-methoxy-4'-O-5' "-bichalcone (rhuschalcone IV, 4), 4,2',4',4' ',2' ",4' "-hexahydroxy-3,5' "-dihydrochalcone-chalcone (rhuschalcone V, 5), and 4,2',4',4' ',2' ",4' "-hexahydroxy-3,5' "-bichalcone (rhuschalcone VI, 6), repectively. Also obtained was the known compound rhuschalcone I (1). Their structures were determined by spectroscopic and chemical methods, and for 1-3 by total synthesis. All the bichalcones (1-6) tested exhibited selective cytotoxic activity against the HT29 and HCT-116 colon tumor cell lines. PMID- 12762792 TI - New Sesquiterpenes from Litsea verticillata. AB - Seven new sesquiterpenes, named litseagermacrane (1), 7-epi-eudesm-4(15)-ene 1alpha,6alpha-diol (2), 5-epi-eudesm-4(15)-ene-1beta,6beta-diol (4), litseahumulanes A (6) and B (7), and litseachromolaevanes A (11) and B (12), as well as the known compounds 7-epi-eudesm-4(15)-ene-1beta,6beta-diol (3), eudesm 4(15)-ene-1beta,6alpha-diol (5), octahydro-4-hydroxy-3alpha-methyl-7-methylene alpha-(1-methylethyl)-1H-indene-1-methanol (8), 10-hydroxyl-15-oxo-alpha-cadinol (9), and aphanamol II (10), were isolated from an anti-HIV fraction of the leaves and twigs of Litsea verticillata Hance (Figure 1). Isolates 1, 4, and 12 were found to inhibit HIV-1 replication in a green fluorescent protein (GFP)-based reporter cell line (HOG.R5) with IC(50) values of 6.5 (27.5), 17.4 (73.1), and 28.0 (119.7) microg/mL (microM), respectively. The structures of these isolates were determined by spectral data including 1D and 2D NMR spectra. Compound 11 was confirmed by X-ray crystallographic analysis. PMID- 12762791 TI - Marine natural products as novel antioxidant prototypes. AB - Pure natural products isolated from marine sponges, algae, and cyanobacteria were examined for antioxidant activity using a 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) solution-based chemical assay and a 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) cellular-based assay. The DCFH system detects only antioxidants that penetrate cellular membranes. Potent antioxidants were identified and the results from each system compared. The algal metabolites cymopol (1), avrainvilleol (3), and fragilamide (4), and the invertebrate constituent puupehenone (5) showed strong antioxidant activity in both systems. Several compounds were active in the DPPH assay but significantly less active in the DCFH system. The green algal metabolite 7-hydroxycymopol (2) was isolated from Cymopolia barbataand its structure determined. Compound 2 was significantly less active in the DCFH system than cymopol (1). The sponge metabolites (1S)-(+) curcuphenol (6), aaptamine (7), isoaaptamine (8), and curcudiol (9) and the cyanobacterial pigment scytonemin (10) showed strong antioxidant activity in the DPPH assay, but were relatively inactive in the DCFH system. Thus, cellular uptake dramatically affects the potential significance of antioxidants discovered using only the DPPH assay. The apparent "proantioxidants" hormothamnione A diacetate (11) and Laurencia monomer diacetate (12) require metabolic activation for antioxidant activity. Significant advantages are achieved using both a solution- and cellular-based assay to discover new antioxidants. PMID- 12762793 TI - New antimalarial bis-dehydroaporphine alkaloids from Polyalthia debilis. AB - Four new dimeric aporphinoids named bidebilines A-D (1-4), bis-7,7' dehydroanonaine (1), 7-dehydroanonaine-7'-dehydro-8'-methoxyanonaine (2), bis 7,7'-dehydro-8,8'-dimethoxyanonaine (3), and bis-7,7'-dehydro-10,10' dimethoxyanonaine (4), were isolated from the roots of Polyalthia debilis. Their structures were established on the basis of spectral evidence. Compounds 3 and 4 exhibited moderate antimalarial activity with IC(50) values of 5.4 and 4.1 microg/mL, respectively. PMID- 12762794 TI - Tasipeptins A and B: new cytotoxic depsipeptides from the marine cyanobacterium Symploca sp. AB - Two new depsipeptides have been isolated from a Symploca sp. collected in Palau. The gross structures of tasipeptins A (1) and B (2) were determined by standard spectroscopic techniques, and the absolute configuration of the amino acid units was determined by chiral HPLC. The relative stereochemistry of the 3-amino-6 hydroxy-2-piperidone (Ahp) moiety in both structures was determined by analysis of (2,3)J(H,H) values. Oxidation with PCC and acid hydrolysis unmasked this latent glutamic acid moiety, allowing for elucidation of the total configuration of 1 and 2. Tasipeptins A (1) and B (2) were cytotoxic toward KB cells with IC(50) values of 0.93 and 0.82 microM, respectively. PMID- 12762796 TI - Euphane and tirucallane triterpenes from the roots of Euphorbia kansui and their in vitro effects on the cell division of Xenopus. AB - Four new euphane-type triterpenes, kansenone (1), kansenonol (3), 11-oxo kansenonol (4), kansenol (5), and a new tirucallane-type triterpene, epi kansenone (2), were isolated from a 60% EtOH extract of Euphorbia kansui, together with alpha-euphol. Their structures were elucidated on the basis of extensive analysis of their 1D and 2D NMR spectral data. This appears to be the first report of the natural occurrence of euphane/tirucallane-type triterpenes with a ketone at C-7. In vitro treatment of cultured individual Xenopus laevis cells at the blastular stage with 1-4 significantly arrested cleavage of the cells (10 microg/mL of each compound resulted in >50% cleavage arrest). PMID- 12762795 TI - Antidiabetic dimeric guianolides and a lignan glycoside from Lactuca indica. AB - Three novel sesquiterpene lactones, lactucain A (1), B (2), and C (3), and a new furofuran lignan, lactucaside (4), were isolated from Lactuca indica along with nine known compounds, 11beta,13-dihydrolactucin, cichoriosides B, quercetin, quercetin 3-O-glucoside, rutin, apigenin, luteolin, luteolin 7-O-glucuronide, and chlorogenic acid. Among these compounds, latucain C (3) and lactucaside (4) showed significant antidiabetic activity. PMID- 12762797 TI - New bioactive coumarins from Kielmeyera albopunctata. AB - The CH(2)Cl(2) extract of the stem bark of Kielmeyera albopunctata was subjected to a bioassay-linked LC-MS dereplication procedure using the KB cell line to afford the new coumarins 4-(1-methylpropyl)-5,7-dihydroxy-8-(4-hydroxy-3 methylbutyryl)-6-(3-methylbut-2-enyl)chromen-2-one (1), 9-(1-methylpropyl)-4 hydroxy-5-(4-hydroxy-3-methylbutyryl)-2-(1-hydroxy-1-methylethyl)-2,3 dihydrofuro[2,3-f]chromen-7-one (2), and 5,7-dihydroxy-8-(4-hydroxy-3 methylbutyryl)-6-(3-methylbut-2-enyl)-4-phenylchromen-2-one (3). Coumarins 1 and 3 showed moderate cytotoxicity, while 2 was inactive at 20 microg/mL. Compound 1was active in vitro against the trypomastigote form of the parasite Trypanosoma cruzi, killing 80% of the parasites after 24 h contact at 4 degrees C when added at 125 microg/mL to infected murine blood. PMID- 12762798 TI - Absolute stereostructures of new arborinane-type triterpenoids and inhibitors of nitric oxide production from Rubia yunnanensis. AB - The aqueous acetone extract from the roots of a Chinese herbal medicine, Rubia yunnanensis, showed a potent inhibitory effect on nitric oxide production in lipopolysaccharide-activated macrophages. Five new arborinane-type triterpenes, rubianols-a (1), -b (2), -c (3), -d (4), and -e (5), and a new arborinane-type triterpene glycoside, rubianoside I (6), were isolated from the herbal crude extract together with 10 known compounds. The absolute stereostructures of 1-6 were determined on the basis of chemical and physicochemical evidence, including the application of the modified Mosher's method. The effects of the isolated constituents on nitric oxide production in lipopolysaccharide-activated macrophages were examined, and several triterpenes were found to show inhibitory activity. PMID- 12762799 TI - Steroidal glycosides from the rhizomes of Dioscorea spongiosa. AB - A water extract of the rhizomes of Dioscorea spongiosa, which showed antiosteoporotic activity, was examined, and four new pregnane glycosides, named spongipregnolosides A-D (1-4), and two new cholestane glycosides, named spongiosides A (5) and B (6), were isolated together with 15 known glycosides. Their structures were determined on the basis of spectroscopic analysis and chemical methods. Among the isolated compounds, spongioside A (5), hypoglaucin G (7), methylprotodioscin (8), and (R)-oct-1-en-3-yl O-alpha-l-arabinopyranosyl-(1- >6)-alpha-d-glucopyranoside (9) showed potent inhibition against bone resorption induced by parathyroid hormone in a bone organ culture system. PMID- 12762800 TI - Ulongapeptin, a cytotoxic cyclic depsipeptide from a Palauan marine cyanobacterium Lyngbya sp. AB - Ulongapeptin (1), a cyclic depsipeptide, was isolated from a Palauan marine cyanobacterium Lyngbya sp. The gross structure was elucidated through one dimensional TOCSY experiments and other spectroscopic techniques. The absolute and relative stereochemistry of the beta-amino acid, 3-amino-2-methyl-7-octynoic acid (AMO), in 1 was determined by synthesis of the saturated alpha-alkyl-beta amino acid and Marfey's analysis of the acid hydrolysate of tetrahydro-1. Ulongapeptin (1) was cytotoxic against KB cells at an IC(50) value of 0.63 microM. PMID- 12762801 TI - Plakortides M and N, bioactive polyketide endoperoxides from the Caribbean marine sponge Plakortis halichondrioides. AB - From a small specimen of the marine sponge Plakortis halichondrioides collected in Puerto Rico we have isolated the known unsaturated ester methyl (2Z,6R,8R,9E) 3,6-epoxy-4,6,8-triethyl-2,4,9-dodecatrienoate (1) along with the known cyclic peroxide plakortide F (2). In addition, the structures of two new polyketide endoperoxides, namely, plakortide O (3) and plakortide P (4), were fully characterized by spectroscopic and chemical methods. The absolute stereochemistry of plakortide O methyl ester (3a) has been determined by analysis of the (R)- and (S)-MTPA esters of the acyclic derivative 5 obtained by hydrogenolysis. Plakortide O (3) and plakortide P (4) exhibited potent cytotoxicity in the NCI human cancer screening program, whereas plakortide O methyl ester, 3a, displayed strong antimalarial activity against Plasmodium falciparum [corrected]. PMID- 12762802 TI - Pachyclavulariolides M-R, six novel diterpenoids from a Taiwanese soft coral Pachyclavularia violacea. AB - Six novel diterpenoids, pachyclavulariolides M-R (1-6), have been isolated from a Taiwanese soft coral Pachyclavularia violacea. The structures and relative stereochemistry of compounds 1-6 were established by spectroscopic analyses. Compound 1 has been shown to exhibit significant cytotoxicity toward P-388 cancer cells. Biosynthesis of 4-6 is discussed. PMID- 12762804 TI - Chlorinated acetylenes from the San Diego sponge Haliclona lunisimilis. AB - The sponge Haliclona lunisimilis from Point Loma, California, contained six known chlorinated acetylenes, previously isolated from the dorid nudibranch Diaulula sandiegensis, and three new metabolites, (1Z,3Z)-1-chlorohexadeca-1,3-diene-5,7 diyne-14-ol, (1Z,3E,9Z)-15-acetoxy-1-chlorohexadeca-5,7-diyne-1,3,9-triene, and (1Z,3E)-14-acetoxy-1-chlorohexadeca-1,3-diene-5,7-diyne. The structures of the new compounds were elucidated by interpretation of spectroscopic data. The relationship between the sponge metabolites and the nudibranch metabolites is discussed. PMID- 12762803 TI - Brominated polyacetylenes from the Philippines sponge Diplastrella sp. AB - Five novel brominated polyacetylenic diols, diplynes A-E (2-6), and three sulfated analogues, diplyne A 1-sulfate (7), diplyne C 1-sulfate (8), and 2 deoxydiplyne D sulfate (9), were isolated from the Philippines sponge Diplastrella sp. by employing bioassay-guided fractionation using the HIV-1 integrase inhibition assay. The novel metabolites were characterized by interpretation of spectroscopic data. PMID- 12762805 TI - Apotirucallane and Tirucallane triterpenoids from Luvunga sarmentosa. AB - The leaves of Luvunga sarmentosa yielded two new apotirucallane triterpenoids, 3 epi-skimmiarepin A (1) and 21,23-epoxy-7alpha,21-dihydroxyapotirucalla-14,24-dien 3-one (2), and a new tirucallane triterpene, 3-epi-flindissol (3). Because of a hemiacetal functionality at C-21, all compounds occurred as mixtures of 21 epimers. 3-epi-Skimmiarepin A (1) and 3-epi-flindissol (3) were oxidized to the corresponding gamma-lactones. The structures have been elucidated on the basis of mass and NMR spectroscopic methods. PMID- 12762806 TI - Callyspongenols A-C, new cytotoxic C22-polyacetylenic alcohols from a red sea sponge, Callyspongia species. AB - Investigation of the organic extract of a Red Sea sponge, Callyspongia sp., resulted in isolation and identification of three new C(22)-polyacetylenic alcohols, callyspongenols A-C (1-3), together with dehydroisophonochalynol (4). The structures of 1-3 were determined by 1D and 2D NMR studies and mass spectral determinations. Compounds 1-4 showed moderate cytotoxicity against P388 and HeLa cells. PMID- 12762808 TI - New sesquiterpene quinols from a Micronesian sponge, Aka sp. AB - Three new sesquiterpene quinols (1, 2, and 5) and two known ones (3 and 4) were isolated along with halistanol sulfate (6) from a marine sponge of the genus Aka collected from Yap Island, Federated States of Micronesia. Their structures were determined from spectral data, and the structure of siphonodictyal C (3) was revised. Sulfates 3 and 6 inhibit CDK4/cyclin D1 complexation, whereas 1 and 4 do not. PMID- 12762807 TI - Three new cyclic peroxides from the marine sponge Plakortis aff simplex. AB - In our continuing program to identify bioactive compounds from marine invertebrates, the MeOH-EtOAc (1:1) extract of the sponge Plakortis aff simplex, collected in Madagascar, was found to be cytotoxic to a series of human tumor cells. From this sponge, three new compounds and one known one, two new 1,2 dioxane peroxylactones named plakortolides H (1) and I (2), and one new 1,2 dioxolane, designated andavadoic acid (3), have been isolated and their structures elucidated. In addition, the known N(3)-methyladenine (4) was isolated, to the best of our knowledge, for the first time from a marine source. Andavadoic acid (3) showed significant activity against 13 tumor cells with GI(50) values in the submicromolar range. PMID- 12762809 TI - Acaricidal activity of tonka bean extracts. Synthesis and structure-activity relationships of bioactive derivatives. AB - The acaricidal effects of tonka bean, Dipterix odorata, extracts were investigated on Dermatophagoides pteronyssinus, the European house dust mite, and compared with benzyl benzoate as a standard acaricidal compound. A cyclohexane extract was the most effective, with an EC(50) = 0.075 g/m(2) after a 24 h period, as compared with benzyl benzoate (0.025 g/m(2)). Bioassay-guided fractionation of this extract led to the isolation of coumarin (1). Pharmacomodulation of this compound led us to test 20 analogues (2-21), which were either synthesized or purchased. PMID- 12762810 TI - Sesquiterpene lactones from Lychnophora ericoides. AB - Two new sesquiterpene lactones, (4S,6R,7S,8S,10R,11S,16R)-1-oxo-3(10),8(16) diepoxy-16-methylprop-1Z-enyl-16-methoxygermacra-2-en-6(12)-olide (1) and (4S,6R,7S,8S,10R,11S)-1-oxo-3,10-epoxy-8-angeloyloxygermacra-2-en-6(12)-olide (2), were isolated from Lychnophora ericoides. Their structures, including absolute configuration, were established by spectroscopic methods, including single-crystal X-ray analysis. Monitoring the furanoheliangolide metabolism of L. ericoides revealed an increase in biosynthesis during the plant flowering period. PMID- 12762812 TI - Sesquiterpene lactones from Anthemis altissima and their anti-Helicobacter pylori activity. AB - Seven sesquiterpene lactones, (-) sivasinolide (1), a new naturally occurring eudesmanolide (altissin, 2), desacetyl-beta-cyclopyrethrosin (3), tatridin-A (4), 1-epi-tatridin B (5), 1alpha,10beta-epoxy-6-hydroxy-1,10H-inunolide (6), and spiciformin (7), were isolated from Anthemis altissima. Also isolated were 10 known flavonoids, namely, apigenin (8), kaempferol 4'-methyl ether (9), quercetin (10), quercetin 3-methyl ether (11), isorhamnetin (12), rhamnetin (13), 6 hydroxyquercetin 3,6,4'-trimethyl ether (14), isoquercetrin (15), taxifolin (16), and eriodictyol (17), and one phenolic acid, chlorogenic acid (18). The structure and the stereochemistry of compound 2 were deduced by spectroscopic methods. The in vitro activity of the sesquiterpene lactones (1-5) against Helicobacter pylori, as well as against three Gram-positive and three Gram-negative bacteria growing aerobically, was tested using the microdilution method. Compounds 8-18 have also been tested against H. pylori. PMID- 12762811 TI - Sterol and triterpenoid constituents of Verbena littoralis with NGF-potentiating activity. AB - Two new sterols, stigmast-5-ene 3beta,4beta,7alpha,22alpha-tetraol (1) and stigmast-5-ene 3beta,7alpha,22alpha-triol (2), were isolated from the aerial parts of a Paraguayan medicinal plant, Verbena littoralis, together with the known compounds ursolic acid (3) and oleanolic acid (4). The structures of 1 and 2 were elucidated by spectroscopic analyses. Compounds 2-4 showed an enhancing activity of nerve growth factor (NGF)-mediated neurite outgrowth in PC12D cells. PMID- 12762813 TI - Salvinorins D-F, new neoclerodane diterpenoids from Salvia divinorum, and an improved method for the isolation of salvinorin A. AB - Three new neoclerodane diterpenoids, salvinorins D-F (4-6), have been isolated from the leaves of Salvia divinorum. The structures were elucidated by chemical and spectroscopic methods, particularly 1D and 2D NMR. A simplified isolation method using chromatography on activated carbon also gave improved yields of the controlled substance salvinorin A (1) and of salvinorin C (3). PMID- 12762814 TI - Two new xanthone derivatives from the algicolous marine fungus Wardomyces anomalus. AB - A marine fungal isolate, identified as Wardomyces anomalus, was cultivated and found to produce two new xanthone derivatives, 2,3,6,8-tetrahydroxy-1 methylxanthone (1) and 2,3,4,6,8-pentahydroxy-1-methylxanthone (2), in addition to the known xanthone derivative 3,6,8-trihydroxy-1-methylxanthone (3) and the known fungal metabolite 5-(hydroxymethyl)-2-furanocarboxylic acid (4). The structures of all compounds were determined on the basis of extensive spectroscopic measurements (1D and 2D NMR, MS, UV, and IR). Compounds 1 and 4 showed significant antioxidant activities. The total extract and 1, 3, and 4 were shown to be inhibitors of p56(lck)tyrosine kinase. PMID- 12762815 TI - Antimalarial dihydroisocoumarins produced by Geotrichum sp., an endophytic fungus of Crassocephalum crepidioides. AB - Three novel dihydroisocoumarin derivatives (1-3) with antimalarial, antituberculous, and antifungal activities have been isolated by bioassay-guided fractionation from an endophytic fungus, Geotrichum sp., collected from Crassocephalum crepidioides. Structures were established as 7-butyl-6,8-dihydroxy 3(R)-pent-11-enylisochroman-1-one (1), 7-but-15-enyl-6,8-dihydroxy-3(R)-pent-11 enylisochroman-1-one (2), and 7-butyl-6,8-dihydroxy-3(R)-pentylisochroman-1-one (3) using spectroscopic data. PMID- 12762816 TI - Isolation of homodolastatin 16, a new cyclic depsipeptide from a Kenyan collection of Lyngbya majuscula. AB - An examination of an organic extract of the cyanobacterium Lyngbya majuscula, collected from Wasini Island off the southern Kenyan coast, led to the isolation of the known cyclic depsipeptide antanapeptin A (1), recently isolated from a Madagascan collection of L. majuscula, and a new bioactive cyclic depsipeptide, homodolastatin 16 (2). The structures of these two compounds were determined from NMR and mass spectrometry data. Homodolastatin 16, a higher homologue of the potential anticancer agent dolastatin 16, exhibited moderate activity against oesophageal and cervical cancer cell lines. PMID- 12762817 TI - Structure reassignment of the fungal metabolite TAEMC161 as the phytotoxin viridiol. AB - Two-dimensional NMR analyses including HMBC, NOESY, and ROESY as well as 1D NOE experiments led to a reassignment of the structure of the recently identified Trichoderma hamatum metabolite TAEMC161 (1) as the previously known viridiol (2). In addition, GIAO-calculated (13)C NMR chemical shifts of 1 and 2 provided strong support for the revised structure. PMID- 12762818 TI - A new antifouling hexapeptide from a Palauan sponge, Haliclona sp. AB - Bioassay-guided fractionation of an extract of the sponge Haliclona sp. provided a known hexapeptide, waiakeamide (1), and a new sulfone derivative (2). The structures of hexapeptides 1 and 2 were elucidated by extensive NMR analyses and the advanced Marfey's method with LC/MS. These compounds showed repellent activity against the blue mussel, Mytilus edulis galloprovincialis. PMID- 12762819 TI - Acetylenic acids from the aerial parts of Nanodea muscosa. AB - The aerial parts of Nanodea muscosa, collected in Chile, yielded two new acetylenic acids. Their structures were elucidated by spectroscopic analyses, including 2D NMR techniques, as (13E)-octadec-13-en-11-ynoic acid (1) and (2E) octadec-2-en-4-ynedioic acid (2). Compound 2 constitutes the first example of a conjugated ene-yne fatty diacid isolated from a natural source. Compounds 1 and 2 did not exhibit toxicity toward a panel of DNA damage checkpoint defective yeast mutants or show affinity for the 5-HT(1A), 5-HT(2A), D(2), and H(1) receptors. PMID- 12762820 TI - New lysophosphatidylcholines and monoglycerides from the marine sponge Stelletta sp. AB - Two new lysophosphatidylcholines (1, 2) and four new monoglycerides (5-8) were isolated from the marine sponge Stelletta sp. by bioactivity-guided fractionation. The planar structures of the new compounds were established on the basis of NMR and MS analyses. The stereochemistry was defined by comparison of the optical rotation. The compounds were evaluated for cytotoxicity against a small panel of five human tumor cell lines. PMID- 12762821 TI - Isolation and synthesis of a new bioactive ellagic acid derivative from Combretum yunnanensis. AB - A new ellagic acid derivative (1) was isolated from the branches of Combretum yunnanensis, and its structure was assigned as 4-(4' '-O-acetyl-alpha rhamnopyranosyl)ellagic acid by analysis of its spectral data. Total synthesis of 1 was achieved by the glycosylation of 3,3'-di-O-benzylellagic acid (5) with 4-O acetyl-2,3-di-O-benzyl-l-rhamnose (4) as a key reaction, and the absolute configuration of 1 was determined. Compound 1 showed weak inhibitory activity against the growth of various tumor cells and inhibited HIV-1 protease. PMID- 12762825 TI - Warm up II: performance changes following active warm up and how to structure the warm up. AB - While warm up is considered to be essential for optimum performance, there is little scientific evidence supporting its effectiveness in many situations. As a result, warm-up procedures are usually based on the trial and error experience of the athlete or coach, rather than on scientific study. Summarising the findings of the many warm-up studies conducted over the years is difficult. Many of the earlier studies were poorly controlled, contained few study participants and often omitted statistical analyses. Furthermore, over the years, warm up protocols consisting of different types (e.g. active, passive, specific) and structures (e.g. varied intensity, duration and recovery) have been used. Finally, while many studies have investigated the physiological responses to warm up, relatively few studies have reported changes in performance following warm up. The first part of this review critically analyses reported changes in performance following various active warm-up protocols. While there is a scarcity of well-controlled studies with large subject numbers and appropriate statistical analyses, a number of conclusions can be drawn regarding the effects of active warm up on performance. Active warm up tends to result in slightly larger improvements in short-term performance (<10 seconds) than those achieved by passive heating alone. However, short-term performance may be impaired if the warm-up protocol is too intense or does not allow sufficient recovery, and results in a decreased availability of high-energy phosphates before commencing the task. Active warm up appears to improve both long-term (>/=5 minutes) and intermediate performance (>10 seconds, but <5 minutes) if it allows the athlete to begin the subsequent task in a relatively non-fatigued state, but with an elevated baseline oxygen consumption (VO(2)). While active warm up has been reported to improve endurance performance, it may have a detrimental effect on endurance performance if it causes a significant increase in thermoregulatory strain. The addition of a brief, task-specific burst of activity has been reported to provide further ergogenic benefits for some tasks. By manipulating intensity, duration and recovery, many different warm-up protocols may be able to achieve similar physiological and performance changes. Finally, passive warm-up techniques may be important to supplement or maintain temperature increases produced by an active warm up, especially if there is an unavoidable delay between the warm up and the task and/or the weather is cold. Further research is required to investigate the role of warm up in different environmental conditions, especially for endurance events where a critical core temperature may limit performance. PMID- 12762824 TI - Plasma leptin and exercise: recent findings. AB - It is established that plasma leptin is associated with satiety and that leptin stimulates lipid metabolism, and increases energy expenditure. These effects implicate leptin as a major regulator of energy homeostasis, which may serve to limit excess energy storage. As plasma leptin concentrations are tightly coupled with fat mass in humans, decreases in adipose mass with weight loss coincide with decreased concentrations of circulating leptin. However, due to many confounding factors, the effects of exercise on circulating leptin are less clear. The data from investigations examining single exercise bouts suggest that serum leptin concentrations are unaltered by short duration (41 minutes or less), non exhaustive exercise, but may be affected by short duration, exhaustive exercise. More convincingly, studies investigating long duration exercise bouts indicate that serum leptin concentrations are reduced with exercise durations ranging from one to multiple hours. These findings raise speculation that exercise-associated reductions in leptin may be due to alterations in nutrient availability or nutrient flux at the level of the adipocytes, the primary site of leptin production and secretion. Thus, one purpose of this review is to discuss the effects of exercise on circulating leptin concentrations with special emphasis on studies that have examined single exercise bouts that are associated with high levels of energy expenditure and energy deficit. In addition, a 'nutrient sensing pathway' (the hexosamine biosynthetic pathway), which regulates leptin gene expression, will be discussed as a possible mechanism by which exercise-induced energy deficit may modulate serum leptin concentrations. PMID- 12762826 TI - Epilepsy in sports and recreation. AB - In the US, millions of people participate in physical activity on a regular basis. However, among the many people with epilepsy, few incorporate exercise into their daily routine. Whether it is because of parental or physician restriction, the fact remains that people with epilepsy are less fit and are not getting the exercise they need. For many years, patients with seizure disorders have been discouraged from participating in physical fitness and team sports due to the fear that it will exacerbate their seizure disorder. However, this overprotective attitude has been slowly changing in light of more recent data on this subject. The evidence shows that patients with good seizure control can participate in both contact and non-contact sports without adversely affecting seizure frequency. This article reviews the effects of exercise on seizure control among patients with epilepsy. It examines the morbidity and mortality associated with exercise, as well as its psychological and physiological effects. Various topics concerning antiepileptic drugs and exercise are also discussed. PMID- 12762830 TI - Thalidomide in cutaneous lupus erythematosus. AB - For nearly 50 years, thalidomide has struggled between success and controversy. After causing an epidemic of phocomelia and other birth defects during the 1960s, affecting thousands of neonates, thalidomide was used as a sedative in selective disorders including leprosy. The potent anti-inflammatory properties of thalidomide were serendipitously discovered while treating patients with erythema nodosum leprosum, and the drug is now approved by the US FDA for the treatment of this disease. Subsequently, the immunosuppressant effects of thalidomide, including the complex modulation of many cytokines, have been recognized. One promising application of thalidomide has been the treatment of cutaneous lupus erythematosus. Among the largest series reviewed, the drug has been found to ameliorate cutaneous lupus erythematosus in 90% of patients, on average. Remission is achieved in approximately 15-20% of patients with cutaneous lupus erythematosus at doses between 50-400 mg daily. Contraceptive concerns and the recognized neuropathic effects of thalidomide limit the use of the drug in patients with cutaneous lupus. Physicians who prescribe thalidomide in the US must be registered with the drug manufacturer. With appropriate control of drug access and close physician monitoring, thalidomide provides a needed therapeutic option for the treatment of refractory cases of cutaneous lupus erythematosus. PMID- 12762827 TI - Heart rate monitoring: applications and limitations. AB - Over the last 20 years, heart rate monitors (HRMs) have become a widely used training aid for a variety of sports. The development of new HRMs has also evolved rapidly during the last two decades. In addition to heart rate (HR) responses to exercise, research has recently focused more on heart rate variability (HRV). Increased HRV has been associated with lower mortality rate and is affected by both age and sex. During graded exercise, the majority of studies show that HRV decreases progressively up to moderate intensities, after which it stabilises. There is abundant evidence from cross-sectional studies that trained individuals have higher HRV than untrained individuals. The results from longitudinal studies are equivocal, with some showing increased HRV after training but an equal number of studies showing no differences. The duration of the training programmes might be one of the factors responsible for the versatility of the results.HRMs are mainly used to determine the exercise intensity of a training session or race. Compared with other indications of exercise intensity, HR is easy to monitor, is relatively cheap and can be used in most situations. In addition, HR and HRV could potentially play a role in the prevention and detection of overtraining. The effects of overreaching on submaximal HR are controversial, with some studies showing decreased rates and others no difference. Maximal HR appears to be decreased in almost all 'overreaching' studies. So far, only few studies have investigated HRV changes after a period of intensified training and no firm conclusions can be drawn from these results. The relationship between HR and oxygen uptake (VO(2)) has been used to predict maximal oxygen uptake (VO(2max)). This method relies upon several assumptions and it has been shown that the results can deviate up to 20% from the true value. The HR-VO(2) relationship is also used to estimate energy expenditure during field conditions. There appears to be general consensus that this method provides a satisfactory estimate of energy expenditure on a group level, but is not very accurate for individual estimations. The relationship between HR and other parameters used to predict and monitor an individual's training status can be influenced by numerous factors. There appears to be a small day-to-day variability in HR and a steady increase during exercise has been observed in most studies. Furthermore, factors such as dehydration and ambient temperature can have a profound effect on the HR-VO(2) relationship. PMID- 12762829 TI - Skin aging and menopause : implications for treatment. AB - The skin is one of the largest organs of the body, which is significantly affected by the aging process and menopause. The significant changes sustained by the skin during the menopause are due to the effect sustained on the skin's individual components. The estrogen receptor has been detected on the cellular components of the skin. Accordingly, dermal cellular metabolism is influenced by the hypoestrogenoemic state of menopause leading to changes in the collagen content, alterations in the concentration of glycoaminoglycans and most importantly the water content. Consequently changes in these basic components leads to an alteration in function compatible with skin aging. Changes in the skin collagen leads to diminished elasticity and skin strength. Collagen content may be measured by various methods such as direct skin biopsy, skin blister assessment for collagen markers and skin thickness measurement. All these variables indicate a reduction in collagen content following menopause. This may be reversed with the administration of estrogen given both topically and systemically.A reduction in hydrophilic glycoaminglycans leads to a direct reduction in water content, which influences the skin turgor. These effects on glycoaminoglycans, due to the hypoestrogenia, have been clearly shown in animal studies and appeared to be rapidly reversed with the application of estrogens. The sum total of these basic effects on the skin leads to wrinkles, the skin condition typifying skin aging.Structures resident in the skin are likewise influenced by menopause. Changes to the cutaneous vascular reactivity are noted following menopause. Capillary blood flow velocity decreases significantly in postmenopausal women. Postmenopausal flushing is due to profound vasodilatation in the dermal papillae. Hair growth is also influenced by the hormonal milieu and consequently hair loss has been associated with the beginning of menopause. Treatments administered for menopause, in particular hormone replacement therapy, appear to alter its effects on the basic components of the skin as well as the more complex structures residing in the skin, consequently retarding the skin aging process. PMID- 12762828 TI - The impact of resistance training on distance running performance. AB - Traditionally, distance running performance was thought to be determined by several characteristics, including maximum oxygen consumption (VO(2max)), lactate threshold (LT), and running economy. Improvements in these areas are primarily achieved through endurance training. Recently, however, it has been shown that anaerobic factors may also play an important role in distance running performance. As a result, some researchers have theorised that resistance training may benefit distance runners. Because resistance training is unlikely to elicit an aerobic stimulus of greater than 50% of VO(2max), it is unlikely that resistance training would improve VO(2max) in trained distance runners. However, it appears that VO(2max) is not compromised when resistance training is added to an endurance programme. Similarly, LT is likely not improved as a result of resistance training in trained endurance runners; however, improvements in LT have been observed in untrained individuals as a result of resistance training. Trained distance runners have shown improvements of up to 8% in running economy following a period of resistance training. Even a small improvement in running economy could have a large impact on distance running performance, particularly in longer events, such as marathons or ultra-marathons. The improvement in running economy has been theorised to be a result of improvements in neuromuscular characteristics, including motor unit recruitment and reduced ground contact time. Although largely theoretical at this point, if resistance training is to improve distance running performance, it will likely have the largest impact on anaerobic capacity and/or neuromuscular characteristics. The primary purpose of this review is to consider the impact of resistance training on the factors that are known to impact distance running performance. A second purpose is to consider different modes of resistance exercise to determine if an optimal protocol exists. PMID- 12762831 TI - A guide to selection and appropriate use of macrolides in skin infections. AB - Dermatologists must be aware of the adverse effects of antimicrobial agents as well as various drug interactions that may influence the choice of drug as well as specific drug schedules. The development of modern antibacterials has improved the treatment of cutaneous bacterial infections. Macrolide antibacterials continue to be an important therapeutic class of drugs with established efficacy in a variety of skin infections. All macrolides inhibit protein synthesis by reversibly binding to the 23S ribosomal RNA in the 50S-subunit. Erythromycin, the prototype of macrolide antibacterials, was isolated from the metabolic products of a strain of Streptomyces erytherus in 1952. Originally, erythromycin was introduced as an alternative to penicillin because of its activity against the Gram-positive organisms. Numerous studies have demonstrated the efficacy and safety of erythromycin for various infectious diseases. Unfortunately, erythromycin is associated with a number of drawbacks including a narrow spectrum of activity, unfavorable pharmacokinetic properties, poor gastrointestinal tolerability, and a significant number of drug-drug interactions. Newer macrolides have been developed to address these limitations. The pharmacokinetics of azithromycin and clarithromycin allow for shorter dosing schedules because of prolonged tissue levels. The efficacy of azithromycin for the treatment of skin and soft tissue infections in adults and children is well established. The unique pharmakinetics of azithromycin makes it a suitable agent for the treatment of acne. Clarithromycin represents a clear advance in the macrolide management of patients with leprosy and skin infections with atypical mycobacteria. Dirithromycin and roxithromycin display no clinical or bacteriological adcantage over erythromycin despite a superior pharmacokinetic profile. An area of concern is the increasing macrolide resistance that is being reported with some of the common pathogens which may limit the clinical usefulness of this class of antimicrobial agents in future. PMID- 12762832 TI - Management of polymorphous light eruption : clinical course, pathogenesis, diagnosis and intervention. AB - Optimal management of patients with polymorphous light eruption (PLE), the most frequent photodermatosis, requires knowledge of the individual clinical course of the disease and pathogenic factors. As PLE often causes problems during leisure time activities and holidays, resulting in a substantial loss of quality of life, prophylaxis is the most important therapeutic approach. Management of PLE must, therefore, focus on basic preventative measures and additional therapeutic approaches, depending on the clinical condition. PLE can be classified into four severity groups (mild, moderate-to-severe, severe and therapy-resistant), which are useful for determining appropriate prophylactic measurements. No specific laboratory tests are available for the diagnosis of PLE, therefore, a clinician must rely on the clinical appearance of the disorder (e.g. clinical symptoms, the location of the lesions, the relationship of the occurrence of the lesions with sun exposure and the time course of the lesions) as well as a patient's medical history in order to make a diagnosis. Basic preventative management of PLE consists of adequate sun protection comprising avoidance of sun exposure, the use of textile sun protection and the application of broadband sunscreens with high UVA protection potential. Other supportive measurements have to be managed individually and are dependent on the patient's medical history and the severity of the disease. Topical antioxidants, systemic immunomodulation, photo(chemo)therapy and systemic immunosuppression may be required in some cases of PLE. Topical antioxidants represent a new treatment approach for moderate-to severe PLE and are an effective and well tolerated option for this patient population. Severe PLE also requires photo(chemo)therapy. Phototherapy can be in the form of 311 nm UVB or UVA1 irradiation. In cases where 311 nm UVB or UVA1 are ineffective, psoralen plus UVA (PUVA) bath therapy may be used. However, PUVA bath therapy must be used with caution because it is associated with acute and long-term adverse effects. In rare exceptions we would consider using oral PUVA therapy. However, in our outpatient department, quality of life of most patients is improved with the treatment regimens that are recommended for patients with moderate-to-severe PLE, without the need for photo(chemo)therapy. PMID- 12762833 TI - Progress in the understanding of the pathology and pathogenesis of cutaneous drug eruptions : implications for management. AB - Cutaneous drug eruptions are among the most common adverse reactions to drug therapy. The etiology may reflect immunologic or nonimmunologic mechanisms, the former encompassing all of the classic Gell and Combs immune mechanisms. Cumulative and synergistic effects of drugs include those interactions of pharmacokinetic and pharmacodynamic factors reflecting the alteration by one drug of the effective serum concentration of another and the functions of drugs and their metabolites that interact to evoke cutaneous and systemic adverse reactions. Recent observations include the role of concurrent infection with lymphotropic viruses and drug effects that, through the enhancement of lymphoid blast transformation and/or lymphocyte survival and the contribution of intercurrent systemic connective tissue disease syndromes, promote enhanced lymphocyte longevity and the acquisition of progressively broadening autoantibody specificities. The latter are particularly opposite to drug-induced lupus erythematosus and to drug reactions in the setting of HIV infection. Specific common types of cutaneous drug eruptions will be discussed in this review. Successful management of cutaneous drug eruptions relies upon the prompt discontinuation of the causative medication; most drug eruptions have a good prognosis after this is accomplished. Oral or topical corticosteroids can be administered to aid in the resolution of some types of eruptions. Antihistamines or anti-inflammatory agents may also be administered for some eruptions. PMID- 12762834 TI - Mycosis fungoides with follicular mucinosis displaying aggressive tumor-stage transformation : successful treatment using radiation therapy plus oral bexarotene combination therapy. AB - Follicular mucinosis is a tissue reaction pattern characterized by mucin deposition with follicular sebaceous units and is found as an idiopathic, primary, benign process (alopecia mucinosa), or as a secondary process due to inflammatory and neoplastic disorders (mycosis fungoides). When associated with follicular mucinosis, mycosis fungoides commonly pursues an aggressive course, often undergoing large-cell transformation, which is associated with resistance to therapy and poor prognosis. We present a case of mycosis fungoides with follicular mucinosis that was treated with incomplete courses of interferon, isotretinoin, and polychemotherapy with subsequent rapid progression to tumor stage mycosis fungoides with large cell transformation and nodal and bone marrow involvement. In this setting, the patient was treated with local radiation therapy, total-skin electron beam therapy, and therapy and maintenance with the oral retinoid-X-receptor retinoid bexarotene, and achieved a durable complete remission. PMID- 12762835 TI - Cutaneous drug reaction case reports: from the world literature. AB - Skin disorders are the most common adverse reactions attributed to drugs. Any skin disorder can be imitated, induced or aggravated by drugs. To help you keep up-to-date with the very latest skin reactions occurring with both new and established drugs, this section of the journal brings you information selected from the adverse drug reaction alerting service Reactions Weekly. The following case reports are selected from the very latest to be published in the world dermatology literature. Any claim of a first report has been verified by a search of AdisBase (a proprietary database of Adis International) and Medline. In addition, the World Health Organization (WHO) Adverse Drug Reactions database is also searched. This database, maintained by the Uppsala Monitoring Centre in Sweden, is the largest and most comprehensive adverse drug reaction source in the world, with information obtained from National Centers of 65 affiliate countries. Each case report is assessed for seriousness using the FDA MedWatch definition of serious (patient outcome is: death; life-threatening; hospitalization; disability; congenital anomaly; or requires intervention to prevent permanent impairment or damage). PMID- 12762837 TI - Lipids in blood-brain barrier models in vitro I: Thin-layer chromatography and high-performance liquid chromatography for the analysis of lipid classes and long chain polyunsaturated fatty acids. AB - The objectives of this study were to optimize a sensitive high-performance liquid chromatography (HPLC) method for fatty acid (FA) analysis for the quantification of polyunsaturated FAs (PUFAs) in cell lipid extracts and to analyze the lipid and FA patterns of three cell lines used in blood-brain barrier (BBB) models: RBE4, ECV304, and C6. Thin-layer chromatographic analysis revealed differences in the phosphatidylcholine-phosphatidylethanolamine (PC:PE) ratios and the triglyceride (TG) content. The PC:PE ratio was <1 for RBE4 cells but >1 for ECV304 and C6 cells. ECV304 cells displayed up to 9% TG depending on culture time, whereas the other cell lines contained about 1% TG. The percentages of docosahexaenoic acid were 9.4 +/- 1.7% of the unsaturated FAs in RBE4 cells (n = 5; 4 d in culture; 9.9% after 10 d), 8.1 +/- 2.0% in ECV304 cells (n = 11; 10 to 14 d), and 6.7 +/- 0.6% in C6 cells (n = 6; 10 to 14 d) and were close to the published values for rat brain microvascular endothelium. The percentage of arachidonic acid (C20:4) was about half that in vivo. ECV304 cells contained the highest fraction of C20:4, 17.8 +/- 2.2%; RBE4 cells contained 11.6 +/- 2.4%; and C6 cells 15.8 +/- 1.9%. It is concluded that a sensitive HPLC method for FAs is now optimized for the analysis of long-chain PUFAs. The results provide a useful framework for studies on the effects of lipid modulation and give reference information for the development of further BBB models. PMID- 12762836 TI - Establishment and characterization of clonal cell lines from the vagina of p53 deficient young mice. AB - Clonal cell lines have been established from vagina of prepubertal female p53(-/ ) mice. Because the mouse vagina has a dual origin (the cranial three-fifths derived from the Mullerian duct and the caudal two-fifths derived from the urogenital sinus), both parts were separately subjected to cloning. Sixteen epithelial and two fibroblastic cell lines were established from the cranial three-fifths (Mullerian vagina group), and four epithelial and three fibroblastic cell lines were established from the caudal two-fifths (sinus vagina group). They were maintained in Dulbecco's modified Eagle medium and Ham's nutrient mixture F 12 containing 10% fetal calf serum and 17 beta-estradiol at 10(-8) M. Two cell lines (one epithelial and one fibroblastic) were examined using soft agar assay, but no colonies were formed. The doubling time of the cell lines was approximately 24 h, and all of them divided more than 200 times without crisis, suggesting that they were immortalized. All epithelial cell lines expressed cytokeratin 8. However, the epithelial cell lines expressed cytokeratin 14 and cytokeratin 10 when exposed to medium containing different concentrations of Ca(2+). Fibroblastic cell lines expressed vimentin. All epithelial and fibroblastic cell lines expressed estrogen receptor-alpha protein. This is the first successful establishment of clonal cell lines from the normal mouse vagina, and these lines may provide good models in vitro of the vagina for the study of the mechanism of estrogen action. PMID- 12762838 TI - Lipids in blood-brain barrier models in vitro II: Influence of glial cells on lipid classes and lipid fatty acids. AB - Lipids of brain tissue and brain microvascular endothelial cells contain high proportions of long-chain polyunsaturated fatty acids (long PUFAs). The blood brain barrier (BBB) is formed by the brain endothelial cells under the inductive influence of brain cells, especially perivascular glia, and coculture of endothelial cells and glial cells has been used to examine this induction. The objective of this study was to investigate whether C6 glioma cells are able to influence the lipid composition and shift the fatty acid (FA) patterns of the BBB model cell lines RBE4 and ECV304 toward the in vivo situation. Lipid classes of the three cell lines were analyzed by thin-layer chromatography and lipid FA patterns by high-performance liquid chromatography. Only ECV304 cells showed altered lipid composition in coculture with C6 cells. The fractions of triglycerides and cholesteryl esters (depending on the support filter) were about twice as high in coculture as when the cells were grown alone. Triglyceride fractions reached 13 to 15% of total lipids in coculture. The three cell lines showed an increase in the percentage of long PUFAs with respect to unsaturated FAs, mainly because of an increase in the percentages of arachidonic acid, all cis-7,10,13,16-docosatetraenoic acid, and all cis-7,10,13,16,19-docosapentaenoic acid. It is concluded that glioma C6 cells are able to induce a more in vivo-like FA pattern in BBB cell culture models. However, changes were not significant for the individual PUFAs, and their levels did not reach in vivo values. PMID- 12762839 TI - Growth of an ovarian cell line of Galleria mellonella and its response to immune inducing factors. AB - Antibacterial proteins are produced in the reproductive tracts of some insect species. The advent of a pupal ovarian cell line of the lepidopteran Galleria mellonella offered an opportunity for exploring the use of ovarian tissue culture to induce antimicrobial proteins in lieu of the larvae. The ovarian cell growth rates and cell yields were maximized by adjusting Grace's medium to pH 6.5, adding 15% (v/v) qualified heat-inactivated fetal calf serum, and lowering the sucrose concentration to 9.3 g/L. Five cell forms and biochemical profiles of the collective cell types were analyzed throughout the culture growth cycle. The final modified culture medium did not affect morphogenesis, whereas it increased the culture growth rate by 50% and the final cell yield threefold. The molting and immunoprotein-inducing hormone, 20-hydroxyecdysone, increased culture growth rate and altered the levels of cell types A and D. Neither 20-hydroxyecdysone nor the larval immunizing agents, apolipophorin-III or Bacillus subtilis, in combination or alone, induced antibacterial activity. The bacterium did induce immunity in both larval and adult stages. PMID- 12762841 TI - Estradiol and a selective estrogen receptor modulator affect steroid hormone receptor messenger RNA levels and turnover in explant cultures of sheep endometrium. AB - Estrogens upregulate estrogen receptor (ER) and progesterone receptor (PR) gene expression in endometrium immediately before ovulation to prepare it for nurturing embryos. Most in vitro model systems have lost the ability to upregulate expression of the ER gene in response to estradiol (E2) or the ability to express the ER gene at all. Here, we used explant cultures from control and E2 treated ewes and assessed expression of four genes (ER, PR, glyceraldehyde 3 phosphate dehydrogenase [GAPDH], and cyclophilin [CYC] genes) that are upregulated by E2 in vivo on Northern blots. In cultures from control and E2 treated ewes, ER and PR messenger ribonucleic acid (mRNA) levels dropped significantly during 24 h of culture in the absence of E2. Glyceraldehyde 3 phosphate dehydrogenase mRNA levels increased 300% in explants from control ewes to match the higher levels in the endometrium of the E2-treated ewe (in vivo and in explant culture). The only effect of E2 in the explant cultures was to prevent the decrease in PR mRNA. The new selective ER modulator, EM-800 (EM), decreased ER and PR mRNA levels in explants from control ewes but upregulated GAPDH and CYC mRNA levels. The EM treatment in vitro mimicked that of E2 by increasing the half life of ER mRNA in endometrial explants. These data illustrate distinct, gene specific effects of the explant culture process, E2, and EM on the expression of endometrial genes. PMID- 12762840 TI - Novel complex integrating mitochondria and the microtubular cytoskeleton with chromosome remodeling and tumor suppressor RASSF1 deduced by in silico homology analysis, interaction cloning in yeast, and colocalization in cultured cells. AB - Availability of the complete sequence of the human genome and sequence homology analysis has accelerated new protein discovery and clues to protein function. Protein-protein interaction cloning suggests multisubunit complexes and pathways. Here, we combine these molecular approaches with cultured cell colocalization analysis to suggest a novel complex and a pathway that integrate the mitochondrial location and the microtubular cytoskeleton with chromosome remodeling, apoptosis, and tumor suppression based on a novel leucine-rich pentatricopeptide repeat-motif-containing protein (LRPPRC) that copurified with the fibroblast growth factor receptor complex. One round of interaction cloning and sequence homology analysis defined a primary LRPPRC complex with novel subunits cat eye syndrome chromosome region candidate 2 (CECR2), ubiquitously expressed transcript (UXT), and chromosome 19 open reading frames 5 (C19ORF5) but still of unknown function. Immuno, deoxyribonucleic acid (DNA), and green fluorescent protein (GFP) tag colocalization analyses revealed that LRPPRC appears in both cytosol and nuclei of cultured cells, colocalizes with mitochondria and beta-tubulin rather than with alpha-actin in the cytosol of interphase cells, and exhibits phase-dependent organization around separating chromosomes in mitotic cells. GFP-tagged CECR2B was strictly nuclear and colocalized with condensed DNA in apoptotic cells. GFP-tagged UXT and GFP-tagged C19ORF5 appeared in both cytosol and nuclei and colocalized with LRPPRC and beta tubulin. Cells exhibiting nuclear C19ORF5 were apoptotic. Screening for interactive substrates with the primary LRPPRC substrates in the human liver complementary DNA library revealed that CECR2B interacted with chromatin associated TFIID-associated protein TAFII30 and ribonucleic acid splicing factor SRP40, UXT bridged to CBP/p300-binding factor CITED2 and kinetochore-associated factor BUB3, and C19ORF5 complexed with mitochondria-associated NADH dehydrogenase I and cytochrome c oxidase I. C19ORF5 also interacted with RASSF1, providing a bridge to apoptosis and tumor suppression. PMID- 12762843 TI - Methods of horizontal gene transfer determination using phylogenetic data. AB - A new approach for comparative analysis of multiple trees reconstructed for representative protein families is proposed. This approach is based on the hypothesis of gene duplication, gene loss and horizontal gene transfer and makes use of stochastic methods and optimization. We present a species tree of 40 prokaryotic organisms obtained by our algorithm on the basis of 132 clusters of orthologous groups of proteins (COGs) from the GenBank of the National Center for Biotechnology Information (USA). We also present a computer technology intended to determine horizontally transferred genes. Some application results of the technology, based on comparative analysis of protein and species trees, are given. PMID- 12762842 TI - Bioinformatics classification and functional analysis of PhoH homologs. AB - PhoH protein is a putative ATPase belonging to the phosphate regulon in Escherichia coli. EC-PhoH homologs are present in different organisms, but it is not clear if they are functionally related, besides nothing is known about their regulation. To distinguish true functional orthologs of EC-PhoH in different classes of bacteria and to identify their functional role in bacterial metabolic network we performed phylogenetic analysis of these proteins and comparative study of position and regulation of the related genes. Three groups of proteins were identified. Proteins of the first group (BS-PhoH orthologs) are present in most of bacteria and are proposed to be functionally linked to phospholipid metabolism and RNA modification. Proteins of the second group (BS-YlaK orthologs) are present in most of aerobes and Actinobacterial YlaK orthologs are shown to be members of a fatty acid beta-oxidation regulons. EC-PhoH orthologs are classified in a third group, specific for Enterobacteria. Functional role of PhoH homologs in the lipid and RNA metabolism and proposed interrelation of PhoH paralogs in one organism are discussed. PMID- 12762844 TI - An algorithm for identification of regulatory signals in unaligned DNA sequences, its testing and parallel implementation. AB - We describe an algorithm (IRSA) for identification of common regulatory signals in samples of unaligned DNA sequences. The algorithm was tested on randomly generated sequences of fixed length with implanted signal of length 15 with 4 mutations, and on natural upstream regions of bacterial genes regulated by PurR, ArgR and CRP. Then it was applied to upstream regions of orthologous genes from Escherichia coli and related genomes. Some new palindromic binding and direct repeats signals were identified. Finally we present a parallel version suitable for computers supporting the MPI protocol. This implementation is not strictly bounded by the number of available processors. The computation speed linearly depends on the number of processors. PMID- 12762845 TI - Two genetic algorithms for identification of regulatory signals. AB - There exist numerous algorithms for identification of regulatory signals in unaligned DNA fragments. Here we present two genetic algorithms for signal identification and describe their implementation and testing on simulated and real data. The first algorithm selects the start position of the signal in a given fragment. The second one builds a "universal" word that is recognized by the transcription factor. We compare these approaches and study the behavior of the genetic algorithm. PMID- 12762846 TI - Combining genome and mouse knockout expression data to highlight binding sites for the transcription factor HNF1alpha. AB - The identification of regulatory elements in silico is an important method for inferring function from sequence data, but it is uncertain which methods are best. We used a novel combination of expression data from a TCF1 knockout mouse (TCF1 codes for the transcription factor HNF1a), and human and mouse genome sequences, to search 2kb upstream of 28 genes downregulated in TCF1 null mice compared to wild type mice. We wrote software (http://www.BindGene.org) to search for and assign p-values to potential binding sites. This identified 8 genes as candidates for being directly regulated by HNF1a: LIPC, CRP, F13B, PRODH2, HSD17B2, SCL7A9, SLC16A7, PAH. There was evidence for conservation between human and mouse for all these regions identified as containing putative binding sites. For three of the genes identified there was experimental evidence for an HNF1a binding site. For comparison we also examined 25 genes up-regulated in TCF1 null mice; only one gene was selected and there was little evidence for conservation of this putative binding site between human and mouse. This result was consistent with HNF1a being a gene transcription activator. Another 6 up-regulated genes had unexpectedly high p-values, suggesting that possibly HNF1a sites have been suppressed from these genes. In conclusion, gene expression data from transgenic animals lacking a transcription factor can be used to identify DNA binding sites for that factor. PMID- 12762847 TI - Prediction of potential C/EBP/NF-kappaB composite elements using matrix-based search methods. AB - Bacterial infections trigger a wide range of host cell responses. For the interaction of Pseudomonas aeruginosa and epithelial cells it is known that transcription factor NF-kappaB plays a central role, but its effects have to be specified by cooperation with additional factors. NF-B containing composite elements, e. g. with C/EBP, may be appropriate indicators for new antibacterial response genes. We refined matrix-based search methods for C/EBP, which was necessary because of weak consensi of the previously existing C/EBP matrices, established a model for C/EBP/ NF-kappaB composite element, used it for scanning all known human 5'-flanking sequences and identified 135 new candidate genes. The newly constructed C/EBP binding patterns will be available with one of the next releases of the TRANSFAC database (http://www.gene-regulation.de). PMID- 12762848 TI - Recognition of eukaryotic promoters using a genetic algorithm based on iterative discriminant analysis. AB - A new approach to recognizing promoter regions of eukaryotic genes is proposed and illustrated by an example of Drosophila melanogaster. The essence of its novelty is in realizing the genetic algorithm to search for optimal partition of a promoter region into local nonoverlapping fragments and selection of the most significant dinucleotide frequencies for the fragments obtained. The method developed was applied to recognizing TATA-containing (TATA+) and DPE-containing (DPE+) promoters of Drosophila melanogaster genes. The program for promoter recognition is included into the GeneExpress system, section RegScan (http://wwwmgs.bionet.nsc.ru/mgs/programs/proga/). PMID- 12762849 TI - Making the body plan: precision in the genetic hierarchy of Drosophila embryo segmentation. AB - We quantify fluctuations in protein expression for three of the segmentation genes in the fruit fly, Drosophila melanogaster. These proteins are representative members of the first three levels of a signalling hierarchy which determines the segmented body plan: maternal (Bicoid protein); gap (Hunchback protein); and pair-rule (Even-skipped protein). We quantify both inter-embryo and inter-nucleus (within a single embryo) variability in expression, especially with respect to positional specification by concentration gradient reading. Errors are quantified both early and late in cleavage cycle 14, during which the protein patterns develop, to study the dynamics of error transmission. We find that Bicoid displays very large positional errors, while expression of the downstream genes, Hunchback and Even-skipped, displays far more precise positioning. This is evidence that the pattern formation of the downstream proteins is at least partially independent of maternal signal, i. e. evidence against simple concentration gradient reading. We also find that fractional errors in concentration increase during cleavage cycle 14. PMID- 12762850 TI - Parametric stability evaluation in computer experiments on the mathematical model of Drosophila control gene subnetwork. AB - Using the method of generalized threshold models, the problem is formulated and solved to evaluate the parametric stability of the model of a gene subnetwork controlling the early ontogenesis of the fruit fly Drosophila melanogaster. Computer experiments have been performed to test the parametric stability of the model. Quantitative evaluations have been obtained for parametric stability of the Drosophila gene subnetwork in nuclei along the embryo's anterior-posterior axis. The results of computer experiments have been compared with the previous research data on "sensitivity" of functioning regimes to random changes of the parameters in the models of prokaryotic and eukaryotic systems, namely the system controlling the lambda-phage development and the subsystem controlling the flower morphogenesis of Arabidopsis thaliana. The obtained results confirm high parametric stability of gene networks that control the development of organisms. PMID- 12762851 TI - Application of filter technology in photomorphogenesis gene network. AB - Light regulates almost all physiological and biochemical processes in plants. Plants react to quantitative and qualitative light characteristics owing to the system of photoreceptors and a branched network for light signal transduction. Comprehensible visual representations of gene networks using of filter technology in the GeneNet system assist understanding the types of the relationships between the components inside the networks and to define their hierarchical structure. PMID- 12762852 TI - Analysis of bacterial RM-systems through genome-scale analysis and related taxonomy issues. AB - Recognition sites for type II restriction and modification enzymes in genomes of several bacteria are recognized as semi-palindromic motifs and are avoided at a significant degree. The key idea of contrast word analysis with respect to RMS recognition sites, is that under-represented words are likely to be selected against. Starting from over- or underrepresented words corresponding to RMS recognition sites in specific clades, the specificity of unknown R-M systems can be highlighted. Among the known restriction enzymes, that are described in the REBASE database of restriction and modification systems, many of their recognition sites are still uncharacterized. Eventually, this motivates studies aimed at assessing horizontal transferring events of RMS in micro-organisms through the analysis of word usage biases in well-determined genomic regions. A probabilistic model is built on a first-order Markovian chain. Statistics on the k-neighborhood of a word is carried out to assess the biological significance of a genomic motif. Efficient word counting procedures have been implemented and statistics are used for the assessment of the significance of individual words in large sequences. On the basis of the set of most avoided words, and in accordance to the IUPAC coding standards, suggestions are made regarding potential recognition sequences. In certain cases, a comparison of avoided palindromic words in taxonomically related bacteria shows a pattern of relatedness of their R M systems. For strengthening this analysis, the primary protein structure of all type II R-M systems known in REBASE have been blasted against the nr-GENBANK database. The combination of these analyses has revealed some interesting examples of possible horizontal transfer events of R-M systems. PMID- 12762853 TI - In silico search for functionally similar proteins involved in meiosis and recombination in evolutionarily distant organisms. AB - Evolutionarily distant organisms have not only orthologs, but also nonhomologous proteins that build functionally similar subcellular structures. For instance, this is true with protein components of the synaptonemal complex (SC), a universal ultrastructure that ensures the successful pairing and recombination of homologous chromosomes during meiosis. We aimed at developing a method to search databases for genes that code for such nonhomologous but functionally analogous proteins. Advantage was taken of the ultrastructural parameters of SC and the conformation of SC proteins responsible for these. Proteins involved in SC central space are known to be similar in secondary structure. Using published data, we found a highly significant correlation between the width of the SC central space and the length of rod-shaped central domain of mammalian and yeast intermediate proteins forming transversal filaments in the SC central space. Basing on this, we suggested a method for searching genome databases of distant organisms for genes whose virtual proteins meet the above correlation requirement. Our recent finding of the Drosophila melanogaster CG17604 gene coding for synaptonemal complex transversal filament protein received experimental support from another lab. With the same strategy, we showed that the Arabidopsis thaliana and Caenorhabditis elegans genomes contain unique genes coding for such proteins. PMID- 12762854 TI - How well does charge reparametrisation account for solvent screening in molecular mechanics calculations? The example of myosin. AB - The rate constant of an enzyme-catalysed reaction is one of the major target properties to understand protein function. Atomic-detail computer simulations can in principle be used to estimate rate constants from the energy profile along the reaction coordinate. For such simulations, molecular mechanics is combined with a quantum description of the reaction process. In molecular mechanics calculations, the electrostatic field is represented by the Coulomb potential of partial atomic charges which have been parametrised for small building blocks in vacuum and transferred to the macromolecule. In aqueous solution, however, the electrostatic interactions are affected by the solvent polarization. While this can be described by numerically solving the Poisson-Boltzmann equation, it is computationally expensive. A simple approximation to this is to optimally reproduce the electrostatic potential in solution by reparametrising the partial atomic charges in such a way that a simple Coulomb potential can still be used. Such a procedure would allow to perform fast calculations of reaction processes in proteins while accounting for the solvent screening effect. Here, this method is tested on myosin, a motor protein that is both an enzyme and exists in very different conformations. PMID- 12762857 TI - iUDB: an object-oriented system for modelling, integration and analysis of gene controlled metabolic networks. AB - The rate constant of an enzyme-catalysed reaction is one of the major target properties to understand protein function. Atomic-detail computer simulations can in principle be used to estimate rate constants from the energy profile along the reaction coordinate. For such simulations, molecular mechanics is combined with a quantum description of the reaction process. In molecular mechanics calculations, the electrostatic field is represented by the Coulomb potential of partial atomic charges which have been parametrised for small building blocks in vacuum and transferred to the macromolecule. In aqueous solution, however, the electrostatic interactions are affected by the solvent polarization. While this can be described by numerically solving the Poisson-Boltzmann equation, it is computationally expensive. A simple approximation to this is to optimally reproduce the electrostatic potential in solution by reparametrising the partial atomic charges in such a way that a simple Coulomb potential can still be used. Such a procedure would allow to perform fast calculations of reaction processes in proteins while accounting for the solvent screening effect. Here, this method is tested on myosin, a motor protein that is both an enzyme and exists in very different conformations. PMID- 12762856 TI - Database of patterns PROF_PAT for detecting local similarities. AB - We have developed PROF_PAT, a database of patterns, constructed for groups of related proteins and designed to maximize representation of amino acid sequences from the SWISS-PROT database. The purpose of the current study was to demonstrate that PROT_PAT is not only as good as known analogs but surpasses them in some features. 10938 new amino acid sequences from the SWISS-PROT bank were compared with patterns constructed for protein families in the PROF_PAT 1.10 bank. The aim of the comparisons was to estimate some threshold values of "Score" parameter to distinguish random similarities from significant ones. From the 10938 new sequences, 638 did not reveal any similarities with PROF_PAT patterns. Cases of found similarities were divided into three sets: 'positive', 'putative' (or 'unknown'), and 'false positive', containing 7719, 2297 and 284 sequences respectively. Using 20 amino acid sequences from the TrEMBL bank that have no descriptions, PROF_PAT demonstrated specificity at a level that was as good as the best-known "secondary" banks. At the same time, its pattern content and variety of included proteins was significantly richer, and its search speed was 3 10 times higher than those of any other protein family bank used for comparison. PMID- 12762858 TI - A database on alternative splice forms on the integrated genetic map service (IGMS). AB - The IGMS is a comprehensive information system that combines the knowledge from genomic sequence, genetic map and genetic disorders databases. This system is updated weekly and focuses on the analysis of EST data. The IGMS identifies UniGene clusters that are differentially expressed in different types of cancer with respect to different reference tissues. The results can be combined with clinical data to asses the potential relevance of specific genes for patient survival or metastatic spread. The second application maps EST with a specific expression profile. Our third application generates a database of alternative splice forms for nine organisms from EST and mRNA sequence data. The results can be used to find splicing patterns specific for certain tissues or tumour types. AVAILABILITY: http://www.bioinf.mdc-berlin.de/igms/. PMID- 12762859 TI - Detecting Cacopsylla pyricola (Hemiptera: Psyllidae) in predator guts using COI mitochondrial markers. AB - Cacopsylla pyricola (Forster) is one of the most important pests of pear in North America, where several native predators have been considered for integrated pest management (IPM) programmes. Two molecular markers of 271 and 188 bp were developed from C. pyricola cytochrome oxidase I (COI) fragments, in order to study the detection of this species in the gut of arthropod predators. Primer sensitivity and the detection period for pear psylla remains in the guts of Anthocoris tomentosus Pericart were determined. The sensitivity threshold was defined at 10-5 dilution of a C. pyricola fifth-instar nymph in all samples. Predator adults were evaluated immediately after ingestion of one to five C. pyricola nymphs (t = 0) and after 2, 4, 6, 8, 16, 24 and 32 h. Detection of the presence of C. pyricola DNA always lasted longer using the shorter fragment and was observed after 32 h of digestion using both markers. The primers amplifying the 188 bp fragment amplified all four psyllid species tested, whereas the primers designed to amplify the 271 bp fragment did so exclusively for C. pyricola and its close relative, Cacopsylla pyri (Linnaeus). Both primers failed to amplify DNA from representative species of the Coccinellidae, Chrysopidae, Hemerobiidae, Anthocoridae, Miridae, Salticidae, Aphididae, Tetranychidae and the Tortricidae, suggesting their suitability for general trophic studies. PMID- 12762860 TI - Sources of semiochemicals mediating host finding in Callosobruchus chinensis (Coleoptera: Bruchidae). AB - Bruchid pests such as Callosobruchus chinensis (Linnaeus) endanger stored legume seeds throughout the tropical belt. The chemical composition of the headspace volatiles from healthy and fourth instar larvae-infested cowpea seeds were identified, characterized, and compared using gas chromatography-mass spectrometry. Y-tube olfactometer bioassays were performed to evaluate the effect of these chemicals on the orientation of conspecific adult females. Analysis of volatiles released from healthy and infested seeds revealed qualitative differences for three out of the 17 compounds identified. Dimethyl disulphide, isobutenyl methyl ketone and methyl trisulphide were found only in the blend emitted from infested but not from healthy seeds. Quantitative differences were apparent for tridecane which was released in larger amounts from infested seeds. While volatiles collected from healthy seeds were attractive to female bruchids, volatiles collected from infested seeds were repellent. To test the hypothesis that the qualitative differences in the chemical composition found may be due to insect-derived components, the volatiles from frass and fourth instar larvae combined were analysed. These volatiles contained both of the sulphides emitted from infested seed but not from healthy seeds. Although a limited induction of volatiles from cowpea seeds cannot be excluded, it is postulated that behavioural differences of the female weevils are largely due to insect-derived semiochemicals. The potential use of such semiochemicals as part of an integrated pest management strategy is discussed. PMID- 12762861 TI - Evidence for host-associated clones of grape phylloxera Daktulosphaira vitifoliae (Hemiptera: Phylloxeridae) in Australia. AB - Grape phylloxera, Daktulosphaira vitifoliae Fitch, is an important pest of grapevines (Vitis vinifera L.) (Vitaceae). Using microsatellite DNA markers it was demonstrated strong associations can exist between D. vitifoliae asexual lineages and vine host type within a vineyard. Also, in excised root bioassays, D. vitifoliae collected from three regions where different genotypic classes predominated showed host-specific differences in life table parameters of reproductive rate and intrinsic rate of increase. Lastly, comparisons of mitochondrial DNA (cytochrome oxidase I) sequences revealed that D. vitifoliae in Australia have paraphyletic origins and fall into two clades partially related to vine host usage. These findings indicate introduction of separate lineages of D. vitifoliae which have close host associations and as such, have important implications for management of this pest in Australia. PMID- 12762862 TI - Insecticide-treated cattle against tsetse (Diptera: Glossinidae): what governs success? AB - The distributions of insecticide-treated cattle from sites in Tanzania and Zimbabwe were assessed from interviews with livestock owners, analysis of secondary livestock data and mapping technologies. The time-course of tsetse control operations at these sites were then simulated using a mathematical model that assumed diffusive movement and logistic growth in fly populations. A simulation of a tsetse control operation in Mudzi district, north-east Zimbabwe, was in accord with observations that the use of insecticide-treated cattle was unable to prevent substantial re-invasion of tsetse from Mozambique, consequent on the patchy distribution of cattle. The simulation was also consistent with the observed efficacy of a 10-km wide barrier of insecticide-treated targets deployed evenly at 4 km/(-2). Simulation of a control operation on Mkwaja Ranch in Tanzania was in accord with the observation that the use of insecticide-treated cattle reduced the tsetse population on the ranch by c. 90%. Insecticide-treated cattle were used to better effect in the Kagera Region of Tanzania. Simulation of this operation predicts that the deployment of 35,000 treated cattle in the area would result in > 99% control of the tsetse population, consistent with the observed decline, by 1-2 orders of magnitude, in cases of trypanosomiasis in the region. The greater success of the Kagera operation was due to the size and shape of the treated area and, particularly, to the restriction of re-invasion to 20% of the perimeter, compared with > 80% on Mkwaja. Simulation was used to assess how tsetse control could have been improved at Mkwaja. The results suggest that splitting herds into smaller, more numerous, units could have achieved some improvement but, in general, the disease problem would not have been solved by the use of insecticide-treated cattle alone. Only by deploying odour-baited targets in ungrazed areas, or in a 1-3-km barrier around the ranch, could substantially better control (99-99.9%) have been achieved. Sensitivity analyses of the Mkwaja simulation showed that the general conclusions were robust to assumptions regarding cattle distribution and the rates of fly movement and growth. Properly managed and appropriately applied insecticide-treated baits are powerful weapons for tsetse control but should not be used without regard to potential levels of re-invasion, consequent largely on considerations of the size and shape of the treatment area and the density and distribution of the baits. PMID- 12762863 TI - Diving beetles (Dytiscidae) as predators of mosquito larvae (Culicidae) in field experiments and in laboratory tests of prey preference. AB - Field experiments were performed in artificial ponds to evaluate how the density of predatory diving beetles (Dytiscidae) would affect the population levels of mosquito larvae (Culicidae). Mosquitoes colonizing the ponds were predominantly species of the genus Culex. In 2000, most of the dytiscids colonizing the ponds were small (Hydroporus spp.), and these predators had no impact on the size of larval mosquito populations, not even in ponds with added dytiscids. In 2001, larger beetles (Ilybius, Rhantus, and Agabus spp.) were more common, and there were significantly fewer mosquito larvae in ponds with the highest numbers of dytiscids. There was a negative correlation between numbers of diving beetles in the ponds and the mean body length of mosquito larvae. In neither year could dytiscid densities be maintained above a certain level owing to emigration. In laboratory tests, there were marked differences between three common dytiscid species in regard to preferences for Daphnia and Culex species as prey: Colymbetes paykulli Erichson chose mosquito larvae more often, whereas both Ilybius ater (De Geer) and I. fuliginosus (Fabricius) preferred Daphnia spp. All of the tested dytiscids consumed large numbers of prey. Since some dytiscid species can efficiently decrease populations of mosquito larvae, they are probably important in the natural control of these dipterans. PMID- 12762864 TI - Predation and prey size choice by the carabid beetle Pterostichus melanarius (Coleoptera: Carabidae): the dangers of extrapolating from laboratory to field. AB - The impact of predation by the generalist carabid beetle Pterostichus melanarius (Illiger) on populations of the field slug Deroceras reticulatum (Muller), and the effects of prey size on the predator-prey interaction, were measured under semi-field conditions. It was hypothesized that environmental heterogeneity would lead to very different patterns of comparative mortality than might be deduced from size choice experiments conducted in the laboratory. Results from outdoor mini-plots, emulating conditions in a field of wheat, demonstrated that P. melanarius significantly reduced numbers of slugs from all size classes, with no apparent preferences. This was in marked contrast to results from earlier laboratory studies, where this beetle fed preferentially on the smallest slugs. The slugs in the mini-plots ranged in size from 2-100 mg and the numbers in the mini-plot reflected the size frequency distribution in the field. Beetles in mini plots containing high densities of slugs increased significantly in weight, in contrast to beetles in mini-plots with low slug density or no added slugs, which did not. Enzyme-linked immunosorbent assays (ELISA), using anti-slug monoclonal antibodies, showed that where there was a higher density of slugs there was more slug protein in the guts of the beetles. It was concluded that environmental heterogeneity probably provided a greater number and diversity of refugia for smaller than for larger slugs, counteracting laboratory-measured size preferences measured in arenas without refugia. These results have implications for a range of ecological studies involving inter- and intra-specific prey size choice, and emphasize the dangers of extrapolating from the laboratory to the field. PMID- 12762865 TI - Performance of Myzus persicae (Hemiptera: Aphididae) clones on different host plants and their host preference. AB - The performance of eighteen clones of Myzus persicae (Sulzer) on pepper and tobacco plants at 20 degrees C and L16:D8 and the choice of young adult apterae between tobacco and pepper leaf-discs were examined. The clones were collected from weeds and peach in two tobacco-growing regions: Katerini, northern Greece and Karditsa, central Greece (only from weeds) and from Lehonia, central eastern Greece where tobacco is not cultivated. All clones did well on both hosts. However, the analysis of data revealed a significant effect of "region / host plant origin" on aphid performance. The mean values of adult weight, intrinsic rate of increase and fecundity of the clones collected in Lehonia and reared on tobacco were significantly lower than the observed values for clones from Katerini and Karditsa. Aphids from Lehonia had significantly higher mean values for developmental time on tobacco than clones from the other regions whereas the opposite was observed when aphids were reared on pepper. Aphids collected in Lehonia performed better on pepper than those originating from the tobacco growing regions. A choice test revealed differences among the clones originating from different regions. Fifty three percent and 43% of aphids from weeds and peach from Lehonia, respectively, chose pepper. By comparison 41.5% and 40.0% of aphids from peach and weeds from Katerini, respectively and 49.5% of aphids from Karditsa preferred tobacco. The results are discussed in relation to host specialization in M. persicae. PMID- 12762866 TI - Natural enemies of three mealybug species (Hemiptera: Pseudococcidae) found on citrus and effects of some insecticides on the mealybug parasitoid Coccidoxenoides peregrinus (Hymenoptera: Encyrtidae) in South Africa. AB - The population density of mealybug species in some South African citrus orchards has increased to pest status in recent years. The characterization of the natural enemy complex and quantification of their contribution to the control of Planococcus citri (Risso), Pseudococcus longispinus (Targioni-Tozzetti) and Pseudococcus calceolariae (Maskell) on Citrus limon (L.) and Citrus reticulata (Blanco) was investigated through intensive sampling. Eight primary and four secondary parasitoids, and two predator species were identified from P. citri and P. calceolariae. Anagyrus pseudococci (Girault) and Coccidoxenoides peregrinus (Timberlake) were the most common species, accounting for 44% and 21% of the total. Of the five primary parasitoids reared from P. longispinus, A. pseudococci and Anagyrus sp. were predominant, comprising 41% and 30%. Nymphal and adult parasitism (range = 0-26% vs. 0-66%) and predation (range = 0-5.6% vs. 0-4.1%) varied significantly between host trees and mealybug species (P < 0.001). The numbers of nymphal instars and adult stages of P. calceolariae and P. longispinus and the nymphal stage of P. citri that were parasitized and killed by predators correlated significantly with the total number of hosts on which they acted (P < 0.01), suggesting a density-dependent association. Laboratory bioassay of nine contact insecticides (methidathion, methomyl, methyl-parathion, parathion, profenofos and prothiofos) against C. peregrinus indicated that all were highly toxic, causing 98-100% mortality in < 6 h of treatment. The IGRs fenoxycarb and triflumuron did not cause significant parasitoid mortality (P > 0.05). However, a mixture of pyriproxyfen and mineral oil caused a marginally significant mortality (P < 0.05). PMID- 12762867 TI - Sex ratio in the aphid parasitoid Aphidius colemani (Hymenoptera: Braconidae) in relation to host size. PMID- 12762868 TI - Survey of Bemisia tabaci (Hemiptera: Aleyrodidae) biotypes in Italy with the description of a new biotype (T) from Euphorbia characias. PMID- 12762870 TI - Microvascular decompression surgery in the United States, 1996 to 2000: mortality rates, morbidity rates, and the effects of hospital and surgeon volumes. AB - OBJECTIVE: Microvascular decompression (MVD) is associated with low mortality and morbidity rates at specialized centers, but many MVD procedures are performed outside such centers. We studied short-term end points after MVD in a national hospital discharge database sample. METHODS: A retrospective cohort study was performed by using the Nationwide Inpatient Sample, 1996 to 2000. RESULTS: The sample included 1326 MVD procedures for treatment of trigeminal neuralgia, 237 for treatment of hemifacial spasm, and 27 for treatment of glossopharyngeal neuralgia, performed at 305 hospitals by 277 identified surgeons. The mortality rate was 0.3%, and the rate of discharge other than to home was 3.8%. Neurological complications were coded in 1.7% of cases, hematomas in 0.5%, and facial palsies in 0.6%, with 0.4% of patients requiring ventriculostomies and 0.7% postoperative ventilation. Trigeminal nerve section was also coded for 3.4% of patients with trigeminal neuralgia, more commonly among older patients (P = 0.08), among female patients (P = 0.03), and at teaching hospitals (P = 0.02). The median annual caseloads were 5 cases per hospital (range, 1-195 cases) and 3 cases per surgeon (range, 1-107 cases). With adjustment for age, sex, race, primary insurance, diagnosis (trigeminal neuralgia versus hemifacial spasm versus glossopharyngeal neuralgia), geographic region, admission type and source, and medical comorbidities, outcomes at discharge were superior at higher-volume hospitals (P = 0.006) and with higher-volume surgeons (P = 0.02). Complications were less frequent after surgery performed at high-volume hospitals (P = 0.04) or by high-volume surgeons (P = 0.01). The rate of discharge other than to home was 5.1% for the lowest-volume-quartile hospitals, compared with 1.6% for the highest volume-quartile hospitals. Volume and mortality rate were not significantly related, but three of the four deaths in the series followed procedures performed by surgeons who had performed only one MVD procedure that year. Length of stay (median, 3 d) and hospital volume were not significantly related. Hospital charges were slightly higher at higher-volume hospitals (P = 0.007). CONCLUSION: Although most MVD procedures in the United States are performed at low-volume centers, mortality rates remain low. Morbidity rates are significantly lower at high-volume hospitals and with high-volume surgeons. PMID- 12762872 TI - Fate of branch arteries after intracranial stenting. AB - OBJECTIVE: One concern with respect to stent procedures performed to treat patients with intracranial lesions is the fate of normal major arterial branches after stents are placed across them. Because most of these lesions occur at vessel bifurcations or at branch points, a normal major branch often arises near the lesion and may be difficult to avoid during stent positioning. The aim of this article is to describe the angiographic outcome of intracranial major branch arteries crossed by a stent in the intracranial circulation. METHODS: We examined the immediate postprocedural cerebral angiograms of the 40 patients who underwent intracranial stenting at the University at Buffalo, Buffalo, NY, between June 1998 and April 2000. In each of 10 patients, the stent was placed across a normal major branch artery. Stents were used to treat aneurysms in seven patients and intracranial stenosis in three patients. The latest cerebral angiogram available was reviewed, and the patency of the major branch arteries was evaluated. RESULTS: The angiographic follow-up period ranged from 4 days to 35 months (mean follow-up, 10 mo). Each of the 10 major branch arteries was patent. No infarcts were associated with the territory of the major branch arteries crossed by the stents, and no patient experienced a related episode of clinical ischemia. Four patients died as a result of causes unrelated to the stenting procedure. The histology of a middle cerebral artery stent that was placed across a lenticulostriate perforator is presented. CONCLUSION: The flexible, low-profile stents used in this study had no angiographically or clinically apparent effect on the major intracranial branches across which they were placed. PMID- 12762871 TI - Long-term electrical capsular stimulation in patients with obsessive-compulsive disorder. AB - OBJECTIVE: Because of the irreversibility of lesioning procedures and their possible side effects, we studied the efficacy of replacing bilateral anterior capsulotomy with chronic electrical capsular stimulation in patients with severe, long-standing, treatment-resistant obsessive-compulsive disorder. METHODS: We stereotactically implanted quadripolar electrodes in both anterior limbs of the internal capsules into six patients with severe obsessive-compulsive disorder. Psychiatrists and psychologists performed a double-blind clinical assessment. A blinded random crossover design was used to assess four of those patients, who underwent continuous stimulation thereafter. RESULTS: The psychiatrist-rated Yale Brown Obsessive Compulsive Scale score was lower in the stimulation-on condition (mean, 19.8 +/- 8.0) than in the postoperative stimulator-off condition (mean, 32.3 +/- 3.9), and this stimulation-induced effect was maintained for at least 21 months after surgery. The Clinical Global Severity score decreased from 5 (severe; standard deviation, 0) in the stimulation-off condition to 3.3 (moderate to moderate-severe; standard deviation, 0.96) in the stimulation-on condition. The Clinical Global Improvement scores were unchanged in one patient and much improved in the other three during stimulation. During the stimulation-off period, symptom severity approached baseline levels in the four patients. Bilateral stimulation led to increased signal on functional magnetic resonance imaging studies, especially in the pons. Digital subtraction analysis of preoperative [(18)F]2-fluoro-2-deoxy-d-glucose positron emission tomographic scans and positron emission tomographic scans obtained after 3 months of stimulation showed decreased frontal metabolism during stimulation. CONCLUSION: These observations indicate that capsular stimulation reduces core symptoms 21 months after surgery in patients with severe, long-standing, treatment-refractory obsessive-compulsive disorder. The stimulation elicited changes in regional brain activity as measured by functional magnetic resonance imaging and positron emission tomography. PMID- 12762873 TI - Surgical clipping may lead to better results than coil embolization: results from a series of 101 consecutive unruptured intracranial aneurysms. AB - OBJECTIVE: Recent reports in the literature have described a significant discrepancy in adverse outcomes between coil embolization (CE; 10%) and surgical clipping (SC; 25%) for the management of unruptured intracranial aneurysms (UIA). This discrepancy led us to analyze our experience. METHODS: In 1996, we designed a prospective study of patients with UIA in which CE was considered the treatment of choice and was performed if the interventional neuroradiologists deemed the aneurysm's fundus-to-neck ratio accessible for CE. SC was performed only if complete CE was unlikely to be achieved or in patients in whom CE already had failed. RESULTS: CE was performed in 38 patients with at least one UIA (41 UIAs, 83% in the anterior circulation). SC was performed in 39 patients with at least one UIA unsuitable for CE (59 UIAs, including 6 after failed CE, 96.5% in the anterior circulation). For CE, the total obliteration rate was 56.1%, the subtotal was 14.6%, and CE failed in 29.3%. There were transient complications in 10% of the cases and permanent complications in 7.5%. Of the 12 failed CE procedures, 7 (58%) were performed for middle cerebral artery aneurysms. For SC, the total obliteration rate was 93.2%, the subtotal was 1.7%, and SC failed (wrapping) in 5.1%. There were transient complications in 16.3% of the patients and permanent complications in 1.7%. The success rate for CE was similar to that for SC only when CE was used for aneurysms with a fundus-to-neck ratio of at least 2.5. CONCLUSION: SC can produce better results than CE in patients with UIA of the anterior circulation. CE as a first-line treatment should be reserved for patients with UIAs with a fundus-to-neck ratio of 2.5 or greater. PMID- 12762874 TI - Patient outcomes after arteriovenous malformation radiosurgical management: results based on a 5- to 14-year follow-up study. AB - OBJECTIVE: Radiosurgery is commonly performed for patients with small to medium sized arteriovenous malformations (AVMs). However, few articles present overall outcomes after one or more radiosurgical procedures, and few data are available for periods longer than 5 years after AVM radiosurgery. METHODS: Between 1990 and 1997, 144 patients underwent AVM radiosurgery and had angiographic follow-up. Of these patients, 112 (78%) had Spetzler-Martin Grade III or greater AVMs; 37 (26%) were located in the basal ganglia, thalamus, or brainstem. Twenty-six patients (18%) underwent repeat radiosurgery. The mean follow-up of 15 patients who died as a result of AVM bleeding or underwent AVM resection after the initial radiosurgery was 22 months (range, 3-47 mo); the mean follow-up of the remaining 129 patients was 86 months (range, 23-169 mo). RESULTS: Excellent (obliteration without deficit, n = 96) or good (obliteration with minor deficit, n = 9) outcomes were achieved in 73% of patients after one or more radiosurgical procedures. Twenty patients (14%) sustained major deficits (n = 15; five had obliteration) or died (n = 5) after radiosurgery. Sixteen patients (11%) had unchanged neurological examinations but persistent arteriovenous shunting. Five patients (4%) required surgery (cystoperitoneal shunting, n = 1; AVM resection, n = 4) at a median of 65 months after radiosurgery because of symptomatic cyst formation or persistent edema. The radiosurgery AVM score correlated with both excellent (R(2) = -0.93, P = 0.003) and excellent or good (R(2) = -0.92, P = 0.004) outcomes. CONCLUSION: The majority of AVM patients are protected from the risk of future hemorrhage and continue their normal daily activities after radiosurgery. Late complications requiring treatment are rare but can occur many years after patients are considered cured of their AVMs. Overall outcomes after AVM radiosurgery seem to be predicted accurately by the described method. PMID- 12762875 TI - Intraoperative measurement of cortical oxygen saturation and blood volume adjacent to cerebral arteriovenous malformations using near-infrared spectroscopy. AB - OBJECTIVE: Both cortical oxygen saturation (SaO(2)) and cortical blood volume (BV) are related to regional cerebral blood flow. Among a series of patients with cerebral arteriovenous malformations, we examined the problem of hyperemia after embolization/resection or resection with the intraoperative application of near infrared spectroscopy. METHODS: Cortical SaO(2) and BV (intracapillary total hemoglobin concentrations) were measured, with near-infrared spectroscopy, in areas adjacent to the arteriovenous malformation for 20 patients, before and after resection. The absolute values for both parameters and postexcision/preexcision ratios (P-P ratios) were determined. Data were correlated with multiple clinical and preoperative angiographic parameters. RESULTS: Before resection, the mean SaO(2) was 49 +/- 16% and the mean BV was 12.65 +/- 7.41 mg/ml. After resection, SaO(2) increased for 17 of 20 patients and BV increased for 18 of 20 patients. The differences between pre- and postexcision values for both parameters were observed to be statistically significant (P < 0.01). The average P-P ratios were 1.5 +/- 0.4 for SaO(2) and 1.7 +/- 0.4 for BV. Correlation between the P-P ratios for the two parameters was good. The initial BV and the P-P ratios for both SaO(2) and BV were exclusively dependent on the extent of preoperative embolization. Postoperatively, two patients developed intracerebral hemorrhage or severe edema. Both demonstrated high P-P ratios for SaO(2) (2.3 and 2.1) and BV (1.6 and 2.1). CONCLUSION: Intraoperative near infrared spectroscopy is able to detect the hyperemic status of adjacent cortex after arteriovenous malformation resection. However, staged therapy with embolization and surgery results in less hyperemia after resection. While two patients with hyperemic complications exhibited very large increases in SaO(2) and BV, we cannot refute the normal-perfusion pressure breakthrough theory. PMID- 12762876 TI - Intraoperative detection of early microvasospasm in patients with subarachnoid hemorrhage by using orthogonal polarization spectral imaging. AB - OBJECTIVE: Changes of major cerebral vessels in patients with subarachnoid hemorrhage (SAH) are well known from routine cerebral angiography. Data on changes in the microcirculation do not exist. This study sought to provide a qualitative and quantitative analysis of the cortical microcirculation after SAH. METHODS: By means of orthogonal polarization spectral imaging, a qualitative and quantitative analysis of cortical microcirculation was performed during aneurysm surgery in 3 patients with an incidental intracerebral aneurysm and 10 patients with SAH. Vessel diameters, red blood cell velocity, and functional capillary density were analyzed before and after the aneurysm was clipped. RESULTS: Initial capillary density in patients with an incidental aneurysm was 91.5 +/- 36.5 cm( 1) (mean +/- standard deviation) compared with 30.5 +/- 13.8 in patients with SAH (P < 0.05). In patients with SAH, capillary density increased significantly to 53.9 +/- 29.1 cm(-1) (P < 0.05) during the operation, as did the frequency of venules with a red blood cell velocity greater than 2 mm/s (P < 0.05). No significant change of arteriolar or venular diameters was observed. However, in patients with SAH, mono- and multisegmental microvasospasms in arterioles were observed, with a reduction of vessel diameters up to 75.1%. CONCLUSION: Orthogonal polarization spectral imaging is a suitable method to study cerebral microcirculation during surgery. In patients with SAH, capillary density is significantly decreased and small arteries and arterioles of the cortical surface exhibit vasospasm that cannot be detected by angiography or transcranial Doppler sonography. These changes may contribute to the initial clinical symptoms and may have an influence on the clinical postoperative course. PMID- 12762877 TI - Brain metastases treated with radiosurgery alone: an alternative to whole brain radiotherapy? AB - OBJECTIVE: Whole brain radiotherapy (WBRT) provides benefit for patients with brain metastases but may result in neurological toxicity for patients with extended survival times. Stereotactic radiosurgery in combination with WBRT has become an important approach, but the value of WBRT has been questioned. As an alternative to WBRT, we managed patients with stereotactic radiosurgery alone, evaluated patients' outcomes, and assessed prognostic factors for survival and tumor control. METHODS: One hundred seventy-two patients with brain metastases were managed with radiosurgery alone. One hundred twenty-one patients were evaluable with follow-up imaging after radiosurgery. The median patient age was 60.5 years (age range, 16-86 yr). The mean marginal tumor dose and volume were 18.5 Gy (range, 11-22 Gy) and 4.4 ml (range, 0.1-24.9 ml). Eighty percent of patients had solitary tumors. RESULTS: The overall median survival time was 8 months. The median survival time in patients with no evidence of primary tumor disease or stable disease was 13 and 11 months. The local tumor control rate was 87%. At 2 years, the rate of local control, remote brain control, and total intracranial control were 75, 41, and 27%, respectively. In multivariate analysis, advanced primary tumor status (P = 0.0003), older age (P = 0.008), lower Karnofsky Performance Scale score (P = 0.01), and malignant melanoma (P = 0.005) were significant for poorer survival. The median survival time was 28 months for patients younger than 60 years of age, with Karnofsky Performance Scale score of at least 90, and whose primary tumor status showed either no evidence of disease or stable disease. Tumor volume (P = 0.02) alone was significant for local tumor control, whereas no factor affected remote or intracranial tumor control. Eleven patients developed complications, six of which were persistent. Nineteen (16.5%) of 116 patients in whom the cause of death was obtained died as a result of causes related to brain metastasis. CONCLUSION: Brain metastases were controlled well with radiosurgery alone as initial therapy. We advocate that WBRT should not be part of the initial treatment protocol for selected patients with one or two tumors with good control of their primary cancer, better Karnofsky Performance Scale score, and younger age, all of which are predictors of longer survival. PMID- 12762878 TI - Incidences of gliomas and meningiomas in Denmark, 1943 to 1997. AB - OBJECTIVE: The objective of this study was to determine the nationwide, population-based incidences of intracranial gliomas and meningiomas (of all grades) during 55 years of monitoring in Denmark. METHODS: On the basis of reports in the Danish Cancer Registry, we calculated age-standardized, period specific incidences and age- and birth cohort-specific incidences, in 5-year age and calendar intervals, for intracranial gliomas and meningiomas. RESULTS: The incidence of gliomas increased 1.7-fold from 1943 to 1947 to 1993 to 1997, whereas the incidence for meningiomas increased 3.9-fold during the same period. CONCLUSION: Based on complete notification to the Danish Cancer Registry, the overall incidences of intracranial gliomas and meningiomas increased during a 55 year period. These increases were observed for all age groups and both sexes. These increases could be explained on the basis of improved diagnoses of these tumors. For gliomas, a maximal annual incidence of approximately 4 cases/100,000 population (World Standard Population) was observed in Denmark at the end of the study period, suggesting that the effects of improved diagnostic tools have reached their maximum with respect to this tumor type. The same was not observed for the incidence of meningiomas, suggesting that perhaps underreporting is still present. PMID- 12762879 TI - Language functional magnetic resonance imaging in preoperative assessment of language areas: correlation with direct cortical stimulation. AB - OBJECTIVE: The aim of this study was to analyze the usefulness of preoperative language functional magnetic resonance imaging (fMRI), by correlating fMRI data with intraoperative cortical stimulation results for patients with brain tumors. METHODS: Naming and verb generation tasks were used, separately or in combination, for 14 right-handed patients with tumors in the left hemisphere. fMRI data obtained were analyzed with SPM software, with two standard analysis thresholds (P < 0.005 and then P < 0.05). The fMRI data were then registered in a frameless stereotactic neuronavigational device and correlated with direct brain mapping results. We used a statistical model with the fMRI information as a predictor, spatially correlating each intraoperatively mapped cortical site with fMRI data integrated in the neuronavigational system (site-by-site correlation). Eight patients were also studied with language fMRI postoperatively, with the same acquisition protocol. RESULTS: We observed high variability in signal extents and locations among patients with both tasks. The activated areas were located mainly in the left hemisphere in the middle and inferior frontal gyri (F2 and F3), the superior and middle temporal gyri (T1 and T2), and the supramarginal and angular gyri. A total of 426 cortical sites were tested for each task among the 14 patients. In frontal and temporoparietal areas, poor sensitivity of the fMRI technique was observed for the naming and verb generation tasks (22 and 36%, respectively) with P < 0.005 as the analysis threshold. Although not perfect, the specificity of the fMRI technique was good in all conditions (97% for the naming task and 98% for the verb generation task). Better correlation (sensitivity, 59%; specificity, 97%) was achieved by combining the two fMRI tasks. Variation of the analysis threshold to P < 0.05 increased the sensitivity to 66% while decreasing the specificity to 91%. Postoperative fMRI data (for the cortical brain areas studied intraoperatively) were in accordance with brain mapping results for six of eight patients. Complete agreement between pre- and postoperative fMRI studies and direct brain mapping results was observed for only three of eight patients. CONCLUSION: With the paradigms and analysis thresholds used in this study, language fMRI data obtained with naming or verb generation tasks, before and after surgery, were imperfectly correlated with intraoperative brain mapping results. A better correlation could be obtained by combining the fMRI tasks. The overall results of this study demonstrated that language fMRI could not be used to make critical surgical decisions in the absence of direct brain mapping. Other acquisition protocols are required for evaluation of the potential role of language fMRI in the accurate detection of essential cortical language areas. PMID- 12762880 TI - Neuropsychological functioning after surgery in children treated for brain tumor. AB - OBJECTIVE: To describe the neuropsychological functioning of children treated with surgery only for localized brain tumors in Dana-Farber Cancer Institute Protocol 92-077. Subsequent reports will describe the neuropsychological functioning of children treated with surgery and stereotactic radiation therapy on Dana-Farber Cancer Institute 92-077. METHODS: The intellectual functioning of 106 patients was evaluated within 3 months after surgery. An in-depth assessment of the neuropsychological functioning, including an impairment index, was conducted for a subset of 77 school-age children (6-16 yr old) across six functional domains. Descriptive statistics were generated; binomial distribution analyses were performed to assess whether the proportion of individuals with impaired performance on each measure exceeded normative expectations. The impairment index assessed whether poor performance was attributable to a few children or reflected the performance of the cohort as a whole. RESULTS: Although the Full Scale IQ was within normative expectations, the Verbal IQ was higher than the Performance IQ with 45% of individuals showing a significant discrepancy (P < 0.01) between these scales. There was an increased prevalence of poor performance for measures of motor output, verbal memory, and visuospatial organization. The distribution of the impairment index indicated moderate impairment across the school-age cohort rather than severe impairment in a few patients. CONCLUSION: The results document a moderate level of neuropsychological morbidity among children with brain tumors before stereotactic radiation therapy, presumably referable to the tumor itself and the surgery. The extent to which stereotactic radiation therapy may increase this burden will be assessed in follow-up studies evaluating the longitudinal neuropsychological data. PMID- 12762881 TI - Basal ganglia neuronal discharge in primary and secondary dystonia in patients undergoing pallidotomy. AB - OBJECTIVE: Basal ganglia neuronal activity in patients undergoing posteroventral pallidotomy (PVP) for the treatment of primary genetic, secondary, or idiopathic dystonia (DYS) was studied to gain a better understanding of the pathophysiology of DYS. METHODS: Intraoperative neurophysiological data recorded from 15 DYS patients were compared with those from 78 patients with Parkinson's disease (PD) who underwent PVP. RESULTS: Putamen neurons in both DYS and PD patients had low discharge rates. Globus pallidus externa (GPe) and globus pallidus interna (GPi) neurons in DYS patients had significantly lower discharge rates and more irregular discharge patterns than in PD patients. GPe and GPi neurons displayed similar discharge rates and patterns in DYS, whereas in PD, the discharge rate of GPe neurons was lower than that of the GPi neurons. The discharge rate and pattern of GPe and GPi neurons in patients whose DYS was ameliorated by PVP were similar to those in DYS patients who did not benefit from PVP. No significant differences in the rate or pattern of neuronal discharge in patients with DYS of different causes were discernible. PVP was most beneficial in patients with primary genetic DYS. Anesthesia with desflurane depressed the discharge rate of the GPe and GPi neurons, particularly in patients with PD. CONCLUSION: Significant differences in the rates and patterns of discharge of GPe and GPi neurons exist in DYS and PD. The findings are discussed with reference to the current model of the functional connections of the basal ganglia. PMID- 12762882 TI - Predictive value of somatosensory evoked potentials for long-lasting pain relief after spinal cord stimulation: practical use for patient selection. AB - OBJECTIVE: Spinal cord stimulation (SCS) has been used for more than 30 years in patients with intractable neuropathic pain, and global success rates have varied from 40 to 70%, according to reported series. Patient selection is currently based on a preliminary percutaneous test, which is useful but invasive, increases the risk of infection, and has yielded false-positive and false-negative results. In this study, we evaluated an alternative method of predicting the effectiveness of SCS before deciding whether to implant laminotomy electrodes-specifically, assessment of neural conduction in the dorsal columns with the use of somatosensory evoked potentials (SSEPs). Thus, we examined the value of preoperative central conduction time (CCT) of SSEPs to stimulation at the level of the painful area as a possible predictor of patient outcome after SCS. METHODS: Ninety-five patients were evaluated during a mean follow-up period of 18.8 months. Patients were classified into four categories according to the location of the lesion responsible for pain: 28 patients had lesions of the peripheral nerves, 27 had radicular lesions, 8 had root avulsions, and 32 had cord lesions. The SCS electrode was implanted through an interlaminar opening at the upper part of the painful territory without performing a percutaneous screening test. Clinical and social markers of pain relief (i.e., Visual Analog Scale scores, analgesic drug intake, work status) were evaluated prospectively 2 months after implantation and then annually. RESULTS: The global success rate in our study group, with success defined as at least 50% long-term pain relief, was 54.7% (52 of 95 patients). Statistical analyses showed a clear influence of preoperative CCT on SCS outcome. Thus, the success rate was nil in patients with significantly abnormal CCT, whereas it was 75.4% in patients with normal preoperative SSEPs. Significant differences between the two groups of patients also were observed with regard to medication intake and work status. CONCLUSION: Preoperative SSEPs provide an objective prediction of patient outcome after SCS. We suggest that if a patient's CCT is abolished or significantly altered, the patient should not undergo SCS. PMID- 12762883 TI - Brachial plexus avulsion injury repairs with nerve transfers and nerve grafts directly implanted into the spinal cord yield partial recovery of shoulder and elbow movements. AB - OBJECTIVE: Complete avulsion of the brachial plexus is a devastating injury that primarily affects young adults. The current treatment is based on nerve transfers, which yield very limited recovery. In this study, brachial plexus injuries were repaired with nerve transfers and nerve grafts directly implanted into the spinal cord. METHODS: Eight patients with complete brachial plexus avulsion injuries were surgically treated. Roots or target nerves of the brachial plexus were repaired with peripheral nerve grafts directly implanted into the spinal cord and with extraplexal nerve transfers. RESULTS: Muscle reinnervation was observed for six patients who received spinal implants. Among those patients, one recovered M4 muscle power. Reinnervation was observed only in proximal upper limb muscles. CONCLUSION: Muscle reinnervation through nerve grafts directly implanted into the spinal cord was demonstrated. It seems that the combination of intra- and extradural neurotizations improves the proximal muscle function results. However, the extent of this improvement is limited and, in our opinion, does not justify the use of spinal implants. PMID- 12762884 TI - Glioblastoma and cerebral microvascular endothelial cell migration in response to tumor-associated growth factors. AB - OBJECTIVE: Glioma cell migration is determined by a complex interplay between soluble motogens and extracellular matrix components. Several growth factors are thought to be involved in glioma cell migration; however, little is known about their motogenic potency relative to one another. METHODS: Using modified Boyden chamber assays, we compared the chemotactic effects of scatter factor/hepatocyte growth factor (SF/HGF), transforming growth factor (TGF)-alpha, TGF-beta1, TGF beta2, epidermal growth factor (EGF), fibroblast growth factor (FGF)-1, FGF-2, insulin-like growth factor (IGF)-1, IGF-2, platelet-derived growth factor (PDGF) AA, PDGF-BB, vascular endothelial growth factor (VEGF), pleiotrophin (PTN), and midkine (MK) in concentrations ranging from 1 pmol/L to 50 nmol/L on three different human glioblastoma cell lines. Checkerboard analyses distinguished between chemotaxis and chemokinesis. We further investigated the motogenic effects on human cerebral microvascular endothelial cells and analyzed receptor expression profiles. RESULTS: SF/HGF was the most potent chemotactic factor for all three glioblastoma cell lines, inducing up to 33-fold stimulation of migration. TGF-alpha showed the second strongest effect (up to 17-fold stimulation), and FGF-1 was also chemotactic for all three glioblastoma cell lines analyzed (maximal 4-fold effect). EGF, FGF-2, IGF-1, IGF-2, TGF-beta1, and TGF-beta2 were chemotactic for one or two of the cell lines (2- to 4-fold effects), whereas PDGF-AA, PDGF-BB, VEGF, PTN, and MK had no effect. In contrast, the most potent stimulators of cerebral microvascular endothelial cell migration were PDGF-AA (4-fold) and PDGF-BB (6-fold). CONCLUSION: The expression levels of SF/HGF and TGF-alpha as well as their respective receptors, MET and EGFR, are known to correlate with glioma malignancy grade. The particularly strong motogenic effects of these two growth factors suggest that they could be promising targets for an antimigratory component of glioma therapy, at least in comparison with the 12 other factors that were analyzed. PMID- 12762885 TI - Anatomic landmarks of the glossopharyngeal nerve: a microsurgical anatomic study. AB - OBJECTIVE: Compared with other lower cranial nerves, the glossopharyngeal nerve (GPhN) is well hidden within the jugular foramen, at the infratemporal fossa, and in the deep layers of the neck. This study aims to disclose the course of the GPhN and point out landmarks to aid in its exposure. METHODS: The GPhN was studied in 10 cadaveric heads (20 sides) injected with colored latex for microsurgical dissection. The specimens were dissected under the surgical microscope. RESULTS: The GPhN can be divided into three portions: cisternal, jugular foramen, and extracranial. The rootlets of the GPhN emerge from the postolivary sulcus and course ventral to the flocculus and choroid plexus of the lateral recess of the fourth ventricle. The nerve then enters the jugular foramen through the uppermost porus (pars nervosa) and is separated from the vagus and accessory nerves by a fibrous crest. The cochlear aqueduct opens to the roof of this porus. On four sides in the cadaver specimens (20%), the GPhN traversed a separate bony canal within the jugular foramen; no separate canal was found in the other cadavers. In all specimens, the Jacobson's (tympanic) nerve emerged from the inferior ganglion of the GPhN, and the Arnold's (auricular branch of the vagus) nerve also consisted of branches from the GPhN. The GPhN exits from the jugular foramen posteromedial to the styloid process and the styloid muscles. The last four cranial nerves and the internal jugular vein pass through a narrow space between the transverse process of the atlas (C1) and the styloid process. The styloid muscles are a pyramid shape, the tip of which is formed by the attachment of the styloid muscles to the styloid process. The GPhN crosses to the anterior side of the stylopharyngeus muscle at the junction of the stylopharyngeus, middle constrictor, and hyoglossal muscles, which are at the base of the pyramid. The middle constrictor muscle forms a wall between the GPhN and the hypoglossal nerve in this region. Then, the GPhN gives off a lingual branch and deepens to innervate the pharyngeal mucosa. CONCLUSION: Two landmarks help to identify the GPhN in the subarachnoid space: the choroid plexus of the lateral recess of the fourth ventricle and the dural entrance porus of the jugular foramen. The opening of the cochlear aqueduct, the mastoid canaliculus, and the inferior tympanic canaliculus are three landmarks of the GPhN within the jugular foramen. Finally, the base of the styloid process, the base of the styloid pyramid, and the transverse process of the atlas serve as three landmarks of the GPhN at the extracranial region in the infratemporal fossa. PMID- 12762886 TI - The neurosurgeon as local oncologist: cellular and molecular neurosurgery in malignant glioma therapy. AB - Malignant gliomas are among the most challenging of all cancers to treat successfully, being characterized not only by aggressive proliferation and expansion but also by inexorable tumor invasion into distant brain tissue. Although considerable progress has been made in the treatment of these tumors with combinations of surgery, radiotherapy, and chemotherapy, these efforts have not been curative. Neurosurgeons as oncologists have increasingly turned their attention to therapies on a molecular scale. Of particular interest to neurosurgeons is the ability to deliver therapy locally to the tumor site or to take advantage of existing immunological mediators, enhancing drug concentrations or therapeutic cell numbers while bypassing the blood-brain barrier to maximize efficacy and minimize systemic toxicity. Exciting local-therapy approaches have been proposed for these devastating tumors. In this review, we discuss the potential applications of bioreactors, neural stem cells, immunotherapies, biodegradable polymers, and convection-enhanced drug delivery in the treatment of malignant gliomas. These approaches are at different stages of readiness for application in clinical neurosurgery, and their eventual effects on the morbidity and mortality rates of gliomas among human patients are difficult to ascertain from successes in animal models. Nevertheless, we are entering an exciting era of "nanoneurosurgery," in which molecular therapies such as those discussed here may routinely complement existing surgical, radiological, and chemotherapeutic approaches to the treatment of neuro-oncological disease. The potential to deploy any of a number of eloquently devised molecular therapies may provide renewed hope for neurosurgeons treating malignant gliomas. PMID- 12762887 TI - Mitogenic signaling cascades in glial tumors. AB - Gliomas are primary central nervous system tumors that arise from astrocytes, oligodendrocytes, or their precursors. Gliomas can be classified into several groups according to histological features. A number of genetic alterations have been identified in human gliomas; these generally affect either signal transduction pathways activated by receptor tyrosine kinases or cell cycle growth arrest pathways. These observed genetic alterations are now being used to complement histopathological diagnosis. The aim of the present review is to give a broad overview of the receptor tyrosine kinase signaling machinery involved in gliomagenesis, with an emphasis on the cooperative interaction between receptor tyrosine kinase signaling and the cell cycle-regulatory machinery. Understanding molecular features of primary glial tumors will eventually allow for target selective intervention in distinct glioma subsets and a more rational approach to adjuvant therapies for these refractory diseases. PMID- 12762888 TI - Second tumors after radiosurgery: tip of the iceberg or a bump in the road? AB - OBJECTIVE: Radiosurgery-associated second tumors have been reported in four isolated patients during the past 2 years. In our own experience, we are aware of two additional patients. The purpose of this report is to call attention to this potentially emerging problem. METHODS: A review of the English-language literature concerning patients with radiosurgery-associated second tumors was performed. In addition, we report on two patients in our own practice who were treated in the past year. RESULTS: Four patients were found in the literature, and two additional patients were seen by the authors. Malignant tumors occurred as early as 6 years after radiosurgery. The pathological findings in three of these four malignant tumors were glioblastoma multiforme. Benign tumors developed between 16 and 19 years later. Tumors developed both within the full-dose prescription volume and in the lower-dose periphery. Of interest, three of the six patients experienced complications of the radiosurgery treatment before developing second tumors. CONCLUSION: Although patients will increasingly be reported with second tumors after radiosurgery in the future, the overall incidence seems quite low and should not alter current radiosurgical practice. However, continual surveillance of treated patients should be considered. PMID- 12762889 TI - Human horns: a historical review and clinical correlation. AB - OBJECTIVE: Accounts of bony human horns originating from the cranium are found peppered throughout the early medical literature. This study reviews the extant literature regarding these entities to elucidate their authenticity. METHODS: We reviewed both historical and current literature as well as osteological material from our anatomy laboratories for accounts or observations of bony outgrowths of the calvaria in humans. RESULTS: Human horns seem to be mentioned more frequently in the historical literature and are documented primarily with drawings. Moreover, from early accounts, it is often difficult to distinguish true large bony outgrowths from scalp excrescences. Only two cadaveric specimens from our laboratory were noted to have small anomalous bony protuberances, one on the occiput and one on the frontal bone. CONCLUSION: With the lack of either photographic or extreme dry specimen evidence of such human horns, we would propose that benign calvarial tumors, such as osteomas, may have initiated speculation that such entities, i.e., horns, exist in humans but that scalp lesions, exaggeration, legend, and religious beliefs have historically propagated these entities to a mythical status. In addition, early surgical intervention and changes in nomenclature may have also decreased the frequency of such sightings. Finally, many early descriptions have not been repeated in recent history, even in third-world countries lacking advanced medical care. PMID- 12762890 TI - Functional anatomy of the spine by Avicenna in his eleventh century treatise Al Qanun fi al-Tibb (The Canons of Medicine). AB - The history of spinal surgery is an important part of the spine-related sciences. The development of treatment strategies for spine-related disorders is acquired from the Western literature. In this article, an Eastern physician, Ibn Sina, who is known as Avicenna in the West, and his treatise, Al-Qanun fi al-Tibb (the Canons of Medicine), are presented. Eight chapters of this book regarding the functional neuroanatomy of the spine were reviewed and are presented to give insight into the development of the understanding of spinal anatomy and biomechanics. PMID- 12762891 TI - Clinical evaluation of intraparenchymal Spiegelberg pressure sensor. AB - OBJECTIVE: The Spiegelberg 3-PN intraparenchymal pressure sensor was clinically evaluated. DESCRIPTION OF INSTRUMENTATION: The Spiegelberg intraparenchymal pressure sensor is a low-cost device that uniquely performs regular automatic zeroing in situ throughout the measurement period. OPERATIVE TECHNIQUE: The Spiegelberg sensor was inserted in 87 patients who required intracranial pressure monitoring as part of their routine management. Complications were assessed by postoperative computed tomographic scanning and clinical investigation. The automated zeroing procedure was assessed after implantation of the sensor and during long-term measurement. In five patients, the "gold standard' of intraventricular pressure was measured simultaneously and compared with the intraparenchymal or subdural Spiegelberg 3-PN pressure. EXPERIENCE AND RESULTS: No complications associated with the Spiegelberg sensor were observed. The duration of monitoring ranged from 3 to 28 days (mean, 10 d). In 3 patients, technical problems occurred, and in 84 patients, the pressure measurement was successful, including the automatic zeroing procedures performed by the monitor after insertion and hourly thereafter. The absolute difference between the Spiegelberg reading and the intraventricular pressure was less than +/-3 mm Hg in 99.6% and less than +/-2 mm Hg in 91.3% of readings. An Altman-Bland bias plot revealed good agreement between the two methods, with an average bias of 0.5 mm Hg, but revealed a significant trend toward 10% lower Spiegelberg readings with increasing intracranial pressure of >25 mm Hg. There was no difference between intraparenchymal and subdural locations. CONCLUSION: The Spiegelberg 3-PN sensor was reliable and simple to use. It can be recommended for routine intraparenchymal and subdural pressure measurement at a considerably lower price compared with other tip transducers and has the unique advantage of automated zeroing in vivo. PMID- 12762892 TI - Midaortic syndrome and subarachnoid hemorrhage associated with ruptured middle cerebral artery aneurysm: case report and review of the literature. AB - OBJECTIVE AND IMPORTANCE: We describe the presentation, screening, management, and clinical outcome of a 21-year-old man who sought care for a ruptured middle cerebral artery (MCA) aneurysm and midaortic syndrome (MAS). Only three cases of MAS and intracranial aneurysm rupture have previously been described in the literature. CLINICAL PRESENTATION: Cranial computed tomographic (CT) scanning, cerebral and abdominal angiography, and multislice three-dimensional CT angiography were used to evaluate intracerebral hemorrhage and to assess medically intractable hypertension in the patient. Digital subtraction angiography revealed a right MCA aneurysm, and multislice three-dimensional CT angiography revealed narrowing of the abdominal aorta. INTERVENTION: The patient's right MCA aneurysm was successfully clipped via a right pterional craniotomy. A narrowed abdominal aorta was confirmed by an abdominal aortic angiogram (performed at Day 5 after surgery) and then dilated by using percutaneous transcatheter angioplasty during the same session. The patient was normotensive even without antihypertensive medications. Neurological examination and postoperative cranial CT findings were within normal limits at the last follow-up examination, performed 4 months after the operation. CONCLUSION: Our patient is the first reported case of ruptured MCA aneurysm with MAS in an adult. The most important problem in the management of MAS associated with ruptured intracranial aneurysm is medically intractable hypertension, which may markedly increase the incidence of rebleeding. It is hard to achieve normotension unless the narrowed aorta and its branches are dilated. For these reasons, MAS should be considered in patients with medically intractable hypertension associated with ruptured intracranial aneurysm. PMID- 12762893 TI - Composite pleomorphic xanthoastrocytoma-ganglioglioma presenting as a suprasellar mass: case report. AB - OBJECTIVE AND IMPORTANCE: Composite pleomorphic xanthoastrocytoma (PXA) ganglioglioma (GG) is a recently recognized, rare type of neoplasm that most commonly presents as a temporal seizure focus among male patients less than 30 years of age. This case represents the only reported suprasellar presentation, with the youngest reported age at diagnosis. CLINICAL PRESENTATION: We present the case of a 12-year-old boy with new-onset diplopia who was diagnosed as having a large suprasellar mass, which was presumptively designated a craniopharyngioma on the basis of its clinical and radiological appearance. INTERVENTION: Gross total resection of a well-encapsulated tumor with adjacent cysts was achieved, without postoperative neurological deficits. Frozen-section studies suggested pilocytic astrocytoma; however, the final histological diagnosis was composite PXA-GG. CONCLUSION: Composite PXA-GG, although extremely rare, may present as a pediatric suprasellar mass. The histopathological diagnosis is contingent on the demonstration of distinct coincident PXA and GG components. The prognosis after gross subtotal resection is likely favorable; however, long-term follow-up monitoring is indicated for these rare neoplasms. PMID- 12762894 TI - Neonatal subgaleal hematoma causing brain compression: report of two cases and review of the literature. AB - OBJECTIVE AND IMPORTANCE: Neonatal subgaleal hematomas (SGHs) are infrequent but underdiagnosed collections of blood beneath the galea, often caused by vacuum delivery. With massive bleeding into the subgaleal space, exsanguination and hypovolemic shock can cause death in 20 to 60% of newborn infants. We report the first two known patients with extracranial cerebral compression caused by SGH. Also, the surgical evacuation of neonatal SGH has not been described previously. CLINICAL PRESENTATION: One patient was a full-term boy who was delivered via vacuum extraction after an uncomplicated pregnancy. Within a few hours, he developed an expanding fluid collection of the scalp and disseminated intravascular coagulation and shock requiring intubation, inotropic support, and blood transfusions. His head circumference grew from 33 cm at birth to 42 cm. He became progressively lethargic and developed posturing movements. Computed tomography of the head revealed a massive SGH causing gross overlapping of the cranial sutures and diffuse cerebral edema. The other patient was a full-term boy delivered via cesarean section after an unsuccessful attempt at vacuum extraction and forceps delivery. The initial head circumference was 34 cm. Within a few hours, he developed an expanding fluid collection of the scalp and became progressively lethargic with posturing. Magnetic resonance imaging of the head revealed a massive SGH with cranial compromise and diffuse cerebral edema. INTERVENTION: Both children had radiographic features indicative of elevated intracranial pressure as well as neurological decompensation. The first patient was taken to the operating room, and the hematoma was evacuated through a small scalp incision. Initially, approximately 150 ml of blood was removed, and a Jackson-Pratt drain diverted another 200 ml of blood during the next 2 days. The infant made a good recovery. In the second case, the patient remained too unstable for operative intervention and died. CONCLUSION: Extracranial cerebral compression represents another way by which neonatal SGH may jeopardize the infant's life. Management consists of measures to correct hypovolemic shock and disseminated intravascular coagulation, as well as surgical intervention to control elevated intracranial pressure. PMID- 12762895 TI - Arterial reconstruction by direct surgical clipping of a basilar artery dissecting aneurysm after failed vertebral artery occlusion: technical case report and literature review. AB - OBJECTIVE AND IMPORTANCE: Dissecting aneurysms of the basilar artery are rare lesions with significant morbidity and mortality. Their management is controversial and often difficult. Although the rebleeding rate is high, clip reconstruction carries prohibitive risk because of the damage to the parent vessel induced by the dissection and the lack of tissue to gather. An enlarging pseudoaneurysm in the chronic phase, however, may have sufficient tissue for clip reconstruction. We present a case in which this strategy was used successfully. CLINICAL PRESENTATION: A 45-year-old woman presented 3 months after an initial presentation with a subarachnoid hemorrhage from a dissecting aneurysm of the basilar trunk at an outside institution. The aneurysm had grown compared with previous angiograms. INTERVENTION: The dominant vertebral artery was sacrificed. Despite this, the aneurysm continued to enlarge. Given the progressive enlargement of the aneurysm, the decision was made to proceed with arterial reconstruction by direct surgical clipping of the saccular component of the dissecting aneurysm. The patient made an excellent recovery with a durable result. CONCLUSION: Although clipping an intracranial pseudoaneurysm in the acute phase may carry a prohibitive risk, clipping such an aneurysm in the chronic phase may occasionally be warranted. To our knowledge, this is the first case reported in the literature in which direct surgical clipping was used as the primary mode of treatment for a basilar artery dissecting aneurysm that enlarged despite occlusion of the dominant vertebral artery. We review the literature on this rare pathological entity and discuss our management strategy. PMID- 12762896 TI - Midline ventral intradural schwannoma of the cervical spinal cord resected via anterior corpectomy with reconstruction: technical case report and review of the literature. AB - OBJECTIVE AND IMPORTANCE: Spinal cord schwannomas are intradural nerve sheath tumors that almost universally occupy a dorsolateral, lateral, or ventrolateral position. Therefore, resection of these lesions typically proceeds via a posterior or posterolateral approach. CLINICAL PRESENTATION: We present a case of a midline ventral intradural schwannoma of the cervical spinal cord causing myelopathy. To the best of our knowledge, no previous reports specifically discuss purely midline ventral intradural schwannomas. INTERVENTION: Resection of the tumor was performed via an anterior cervical corpectomy with spinal arthrodesis and fixation. We review possible causes for such an anomalous location for schwannoma as well as the advantages and disadvantages of various surgical strategies for removing the tumor. CONCLUSION: This case exemplifies the usefulness of anterior approaches to the cervical spine in treating unusual intradural spinal cord tumors. PMID- 12762897 TI - Intrasphenoidal encephalocele associated with cerebrospinal fluid fistula and subdural hematomas: technical case report. AB - OBJECTIVE AND IMPORTANCE: Intrasphenoidal encephalocele is a rare clinical entity that is often complicated by rhinorrhea, recurrent meningitis, and headache, but in no case has the association of rhinorrhea with subdural hematomas been described. A surgical procedure to stop persistent cerebrospinal fluid leakage is reported. CLINICAL PRESENTATION: A 59-year-old man sought care for intractable rhinoliquorrhea of 6 months' duration. Cranial computed tomographic and magnetic resonance imaging scans revealed a basal posterior frontal bony defect and an evocative image suggesting intrasphenoidal encephalocele. INTERVENTION: A transnasal transsphenoidal surgical procedure was performed; the encephalocele was removed, and the sphenoid sinus was filled with an inflatable pouch made of synthetic dura mater containing abdominal fat. Postoperative reduction of the rhinoliquorrhea, but not its total disappearance, was observed. Total disappearance was achieved only after endonasal, transmucosal inflation of the pouch with human fibrin glue. One of the subdural hematomas disappeared spontaneously, and the other was treated by a surgical procedure. CONCLUSION: The possible role of the presented technique in the treatment of cerebrospinal fluid leakage is discussed. PMID- 12762898 TI - Endoscopic anatomic features of the triangular recess. AB - OBJECTIVE: To describe the morphological and topographic features of the triangular recess (TR) in the anterior wall of the third ventricle and the pathological conditions that allow its observation during ventricular endoscopic neuronavigation. METHODS: A systematic review of records and operative videotapes for 145 patients who underwent endoscopic third ventriculostomy was performed. RESULTS: The TR could be recognized in five cases of hydrocephalus, each caused by a different underlying pathological condition. The approach was precoronal in four cases and suboccipital in one. The morphological and topographic features of the TR and adjacent structures varied among the different cases. CONCLUSION: Although it is seldom reported in neuroanatomy handbooks and is not readily accessible under normal conditions, the TR is a characteristic structure of the third ventricle, which might become apparent in several conditions that produce hydrocephalus. Neurosurgeons who perform neuroendoscopy should be aware of this structure and of the situations that cause its deformation and allow its observation during endoscopic neuronavigation. PMID- 12762899 TI - Training in microvascular surgery using a chicken wing artery. AB - OBJECTIVE: Microarterial anastomosis is now seldom performed for treatment of atherosclerotic occlusive cerebrovascular disease. However, a small but significant number of procedures still require this technique. When a surgeon's clinical experience is limited, regular practice is required to maintain and improve surgical skills. The present training system involves passage from suturing of synthetic materials (such as Silastic tubes) to practice with experimental living animals or cadavers. However, these methods are neither convenient nor practical for daily exercises and rehearsals. I present a unique training exercise for microarterial anastomosis, using a chicken wing artery. METHODS: A brachial artery can be extracted from a chicken wing. The artery is 5 to 6 cm long and measures approximately 1 mm in diameter. The artery can be used to practice end-to-end, end-to-side, or side-to-side anastomosis under the microscope. RESULTS: Several advantages are noted: the materials are cheap, convenient to manage, and easy to obtain, and neither specific facilities to maintain living animals nor anesthesia is needed. Moreover, the diameter and structure of the material are identical to those of human cortical vessels, making the rehearsal quite similar to the actual surgical experience. CONCLUSION: This exercise is useful not only for young surgeons who wish to learn microsurgical techniques but also for more experienced surgeons who need to maintain or improve their skills. PMID- 12762900 TI - Local electronic storage of radiological studies for radiosurgery. AB - OBJECTIVE: Although radiosurgical practice mandates meticulous radiological follow-up, even the most efficient radiology department can be overwhelmed by the large number of radiosurgical patients who have undergone diagnostic studies for many years at many different institutions to follow many separate lesions. Although the task of assembling these studies is theoretically possible, because they are spread out in time and space, it is often impractical. We therefore sought to construct a computer-based system that could store images from multiple sources and present them instantly for review. METHODS: We attached a flatbed film scanner to a standard desktop computer in our clinic and scanned selected sheets of film into an image database at each visit of a radiosurgical patient. "Low-tech" solutions were deliberately chosen-that is, to enhance ease and software compatibility, we used the operating system's directory structure for organization of data instead of proprietary software. Standard commercially available software was used to review studies that had been previously scanned. RESULTS: During a 2- to 3-year period, images were scanned from 1129 studies performed on 435 patients. Images could be reviewed instantly and compared with current studies, and scanning a single piece of film required approximately 30 seconds. We estimate that the current capacity of our computer memory will satisfy our needs for approximately 12 years. CONCLUSION: Assembly of an efficient and inexpensive system for image storage and retrieval suitable for radiosurgical practice is feasible and straightforward. Although our system is not a substitute for a radiology department, it obviates the constant frustration of "finding the films" and has become an essential part of our radiosurgical practice. PMID- 12762901 TI - Topsy's midlife career in healthcare ethics. PMID- 12762902 TI - The value of bioethics education. PMID- 12762903 TI - Self-directed bioethics education. PMID- 12762904 TI - Why medical humanities? PMID- 12762906 TI - Earl Weaver was right: it's what you learn after you think you know it all that counts. PMID- 12762905 TI - The state of graduate education: one student's view. PMID- 12762907 TI - Why study bioethics? Because it's interesting. PMID- 12762908 TI - Principle and praxis: harmonizing theoretical and clinical ethics. PMID- 12762909 TI - Clearing the mist. PMID- 12762910 TI - The place of philosophy. PMID- 12762911 TI - Research ethics education: the view from below. PMID- 12762912 TI - The meaning of graduate education for bioethics. PMID- 12762914 TI - Now that we have the data, what was the question? PMID- 12762915 TI - A dash of this and a pinch of that: the role of interdisciplinary opportunities in graduate education. PMID- 12762913 TI - A Canadian perspective. PMID- 12762916 TI - Ethics training in graduate medical education. PMID- 12762917 TI - There he is ... master of bioethics. PMID- 12762918 TI - An international student's perspective. PMID- 12762919 TI - The merits of a general education in bioethics. PMID- 12762920 TI - Balancing bioethics. PMID- 12762921 TI - Why not medical humanities? PMID- 12762922 TI - A bull market for biomedical ethics. PMID- 12762923 TI - My bioethics education at Georgetown. PMID- 12762924 TI - The absent professor: why we don't teach research ethics and what to do about it. AB - Research ethics education in the biosciences has not historically been a priority for research universities despite the fact that funding agencies, government regulators, and the parties involved in the research enterprise agree that it ought to be. The confluence of a number of factors, including scrutiny and regulation due to increased public awareness of the impact of basic research on society, increased public and private funding, increased diversity and collaboration among researchers, the impressive success and speed of research advances, and high-profile cases of misconduct, have made it necessary to reexamine how the bioscience research community at all levels provides ethics education to its own. We discuss the need to and reasons for making ethics integral to the education of bioscientists, approaches to achieving this goal, challenges this goal presents, and responses to those challenges. PMID- 12762925 TI - On the importance of research ethics and mentoring. PMID- 12762926 TI - Institutional and individual responsibilities for integrity in research. PMID- 12762927 TI - Ethics in biomedical research: practical considerations. PMID- 12762928 TI - Teaching old dogs new tricks: continuing education in research ethics. PMID- 12762929 TI - Faultlines in "bioscience ethics": lessons from the Human Genome Diversity Project. PMID- 12762930 TI - Addressing the hidden curriculum in scientific research. PMID- 12762931 TI - The professor is excused. PMID- 12762932 TI - Responsible conduct of research is all well and good. PMID- 12762933 TI - Teaching ethics to basic scientists: suggestions for greater curricular clarity. PMID- 12762934 TI - What's the professor got to do with it? PMID- 12762935 TI - It's alive! Giving birth to research ethics education. PMID- 12762936 TI - Cultural synthesis: science and ethics. PMID- 12762952 TI - Intensive diabetes management in pediatric patients. AB - Intensive diabetes management requires frequent home glucose monitoring, multiple daily insulin injections or chronic subcutaneous insulin infusion, and adjustments of insulin doses in response to changes in blood glucose levels, food intake, and exercise. It also requires a periodic review of previous glucose results to recognize patterns of hyper- or hypoglycemia. The goals of intensive management are age dependent. In young children, avoidance of severe hypoglycemia is the major goal. In older children and adolescents, lowering hemoglobin A(1c) becomes an increasingly important goal. In children of all ages, the ability to have a flexible lifestyle and meal plan is often a priority. This article provides a brief overview of the rationale for implementing intensive diabetes management in pediatric patients, and practical guidelines for implementation. PMID- 12762953 TI - Type 2 diabetes in children. AB - The emerging epidemic of type 2 diabetes (T2DM) in young people reflects increasing rates of obesity and parallels the increasing frequency of T2DM in adults. As in adults, T2DM in children is part of the insulin resistance syndrome that includes hyperandrogenism seen as premature adrenarche and polycystic ovary syndrome, hypertension, dyslipidemia, and other atherosclerosis risk factors. Recent studies in children document risk factors for T2DM, and associated cardiovascular risk factors, including obesity, family history, diabetic gestation, and underweight or overweight for gestational age. Genetically determined insulin resistance or limited beta-cell reserve has been demonstrated in high-risk individuals, including first-degree relatives of girls with premature adrenarche. This genetic background, considered advantageous in a feast and-famine existence (the thrifty genotype), is rendered detrimental with abundant food and physical inactivity, a lifestyle demonstrated to be typical of families of children with T2DM. The increasing incidence of T2DM in children and adolescents threatens to become a major public health problem. Risk factors for cardiovascular disease, hypertension, hyperlipidemia, and microalbuminuria are present at diagnosis of T2DM in Native American adolescents, indicating that insulin resistance has been present for some time before the diagnosis of diabetes was made. Case finding is likely to be beneficial in high-risk youths. Treatment is the same as that of adults. The only data on use of oral hypoglycemic agents in children have been with metformin. Community and governmental efforts to educate all children and their parents about the need for physical activity and dietary modification are essential to control this epidemic. PMID- 12762954 TI - Prediction and prevention of type 1 diabetes. AB - The incidence of type 1 diabetes continues to increase worldwide. Despite major strides in the daily care of patients with the disease, the patients' contribution to overall morbidity and mortality statistics and health care economic burden to society is disproportionately large because of the high rate of microvascular and macrovascular complications. The quest for prevention of type 1 diabetes has been made feasible by the unraveling of the immunogenetics of the disease and the identification of at-risk subjects by an enhanced understanding of the natural history of the prediabetic period. A combination of immunologic, metabolic, and genetic markers can be used to accurately predict the disease in higher-risk relatives and the general population. This has enabled initiation of worldwide trials (Diabetes Prevention Trial-Type 1, European Nicotinamide Diabetes Intervention Trial, and Trial to Prevent Diabetes in Genetically at Risk) aimed at the prevention of the disease. Various promising agents are being considered for use in different at-risk populations in the near future. PMID- 12762955 TI - Psychosocial issues in pediatric diabetes. AB - Although the majority of diabetic patients exhibit mild depression, anxiety, and somatic complaints at the time of diagnosis, these symptoms are usually temporary and resolve within 6 to 9 months. However, in some patients, depressive symptoms may increase with the duration of diabetes. Anxiety seems to increase and to be more prevalent in girls than in boys. Depression and self-esteem problems have a negative impact on the adaptation to diabetes and metabolic control. Patients' adjustment to diabetes shortly after diagnosis seems to predict adjustment later on. Family characteristics have major implications in the patient's adjustment to diabetes, self-management, and quality of life. Children and adolescents living in families with a high degree of conflict or that are less caring appear to have poorer metabolic control. Thus, the goal of achieving metabolic and psychological stability requires a diabetes team equipped to provide social and psychological support in addition to the development of technical skills. This includes very early assessment of family dynamics and psychological intervention. PMID- 12762956 TI - Cerebral edema in children with diabetic ketoacidosis. AB - Cerebral edema is the most frequent serious complication of diabetic ketoacidosis (DKA) in children, occurring in 1% to 5% of DKA episodes. The rates of mortality and permanent neurologic morbidity from this complication are high. The pathophysiologic mechanisms underlying DKA-related cerebral edema are unclear. A number of past and more recent studies have investigated biochemical and therapeutic risk factors for the development of cerebral edema. Recent studies have shown that a higher initial serum urea nitrogen concentration and lower initial partial pressure of carbon dioxide are associated with the development of cerebral edema. This and other information suggests that the pathophysiology of DKA-related cerebral edema may involve cerebral ischemia. PMID- 12762957 TI - Complications of pediatric and adolescent type 1 diabetes mellitus. AB - Type 1 diabetes mellitus is potentially associated with serious microvascular and macrovascular complications, although these are usually subclinical during the pediatric and adolescent years. There is no "grace" period for the beginnings of such complications. Duration of diabetes, glycemic control, age, and pubertal stage are critical factors contributing toward development of such problems. Other risk factors include family history (genetic predisposition), hyperlipidemia, hypertension, and smoking. The Diabetes Control and Complications Trial (DCCT) proved the importance of glycemic control and emphasized the ability of improved glucose control to prevent or decrease retinopathy, nephropathy, and neuropathy using a multidisciplinary same-philosophy-of-care approach plus targeted glucose and hemoglobin A(1c) values. Other natural history and intervention studies support the findings of the DCCT. Although our current tools are not perfect, they allow us to decrease microangiopathic complications very significantly if we educate our patients and their family members. Metabolic control counts. PMID- 12762958 TI - New technologies and therapeutic approaches for the management of pediatric diabetes. AB - Evolving trends in technology and therapeutic strategies create a very exciting time for the management of diabetes. Technology, like the internet, helps us to keep up with this fast-paced, changing world of diabetes. New therapies most often begin with adult clinical trials; once safety and efficacy are demonstrated, practice recommendations follow for the pediatric population. The Diabetes Research Working Group has defined potentially important new directions and technologies for diabetes research. The group has recommended the creation of regional centers of technologic excellence, if contemporary diabetes research is to succeed. An overview of recent advances in diabetes technology follows covering five main areas: monitoring, telemedicine, insulin analogues, insulin delivery devices, and islet cell transplantation. PMID- 12762961 TI - Birth weight as a predictor of type 2 diabetes mellitus: the U-shaped curve. AB - Recent findings have sparked intense interest in birth weight as a predictor of type 2 diabetes mellitus. In some populations, there is an inverse association between birth size and disease; however, in the Pima Indians, a population with very high rates of gestational diabetes, high birth weight and low birth weight are associated with diabetes. This results in a U-shaped curve with higher diabetes rates in both tails of the distribution. Similarly, there are reports of an inverse association between stature and prevalence of gestational diabetes, but, in California Latina women with gestational diabetes, there is a positive association between stature and glucose concentration. Could these disparate findings also represent the two ends of a U-shaped curve? The disparities are likely due to differences in the environment, both in utero and postnatally, that different groups experience. PMID- 12762960 TI - Maternal factors that determine neonatal size and body fat. AB - These data are a review of previously published data. Initially, body composition was estimated in 186 neonates. Fat- free mass (FFM), which constituted 86% of birth weight, accounted for 83% of the variance in birth weight; fat mass (FM), which constituted 14% of birth weight, accounted for 46% of the variance in birth weight. Male neonates were an average of 175 g heavier than females. FFM was greater among males compared with females (P = 0.0001). Using stepwise logistic regression, 29% of the variance in birth weight, 30% in FFM, and 17% in FM was accounted for. Independent variables included maternal height, pregravid weight, weight gain during pregnancy, education, parity, paternal height and weight, neonatal sex, and gestational age. Including maternal insulin sensitivity explained 48% of the variance in birth weight, 53% in FFM, and 46% in FM. There was a positive correlation between weight gain and birth weight in control subjects but a negative correlation in subjects with gestational diabetes mellitus. Lastly, the roles of insulin, insulin-like growth factors, and leptin were examined in relation to fetoplacental growth and body composition. The assessment of fetal/neonatal body composition may improve the understanding of the effect of differential factors on fetal growth. Factors associated with accretion of fetal adipose tissue in late gestation are less well understood compared with birth weight and FFM. Additional studies of maternal glucose and lipid metabolism are needed to better evaluate fetal growth. PMID- 12762962 TI - Continuous glucose monitoring during pregnancy complicated by gestational diabetes mellitus. AB - Postprandial glucose has been reported to be the best predictor of neonatal macrosomia. Therefore, self-blood glucose monitoring (SBGM) protocols for diabetic pregnant women stress the importance of measuring blood glucose after meals. However, there is controversy in the literature. Traditionally, glucose monitoring has been prescribed at 2 hours after eating to coincide with the times a patient is at increased risk of hypoglycemia. Human regular insulin peaks at 2 to 3 hours after injection; thus, checking blood glucose at the 2-hour point is a relic of strategies to prevent hypoglycemia. In pregnancy, the emphasis has been on measuring during times when blood glucose levels are highest. One hour after the start of the meal has been shown to be the time of peak postprandial response in 90% of pregnant woman. Because glucose excursions may reach their maximum at varying times, based on the size and number of meals ingested, SBGM alone may not capture the full extent of total daily postprandial hyperglycemia. Because intermittent blood glucose monitoring underestimates the number of hyperglycemic events, a more accurate determination of postprandial glucose levels is necessary to decrease the risk of macrosomia in gestational diabetes mellitus. Continuous glucose monitoring may facilitate the detection of all postprandial peaks, including those due to unscheduled meals, and may provide an opportunity for better intervention by providing the complete glucose profile. PMID- 12762964 TI - Diabetes Prevention in high-risk women with gestational diabetes. PMID- 12762963 TI - The utility of a single test to identify women at risk for gestational diabetes. AB - A number of single tests have been proposed as both screening and definitive tests of glucose intolerance during pregnancy. Despite limitations imposed by a lack of uniformity in methodology and definitions of gestational diabetes mellitus, there appears to be an independent relationship between some single test results and clinically meaningful outcomes. Further study is needed to identify those glucose values above which women and their babies who are at risk for glycemia-related adverse outcomes may be identified. PMID- 12762965 TI - Insulin-mediated capillary recruitment in skeletal muscle: is this a mediator of insulin action on glucose metabolism? AB - The possibility that insulin stimulates microvascular access for itself and glucose in muscle in vivo is discussed. The application of new techniques suggests that capillary recruitment is a normal part of insulin's action and that this process becomes impaired in insulin resistance. Exercise, which also leads to capillary recruitment, may involve a different mechanism than that used by insulin. PMID- 12762966 TI - Surgical removal of visceral adipose tissue: effects on insulin action. AB - Many studies have demonstrated that excess of visceral fat has deleterious effects on insulin action. Mainly, it has been shown to be associated with a decrease in hepatic and peripheral insulin sensitivity, which results in a clinical condition also known as insulin resistance. This report describes a novel experimental method that we employed in order to analyze the particular effects of visceral fat on insulin activity. By extracting visceral fat we were able to distinguish the specific role that it plays in insulin action, and to analyze its effects on the gene expression of a variety of fat-derived peptides, which may be considered to be (at least partially) mediators in the development of the metabolic syndrome and possibly diabetes mellitus. PMID- 12762967 TI - Adiponectin: systemic contributor to insulin sensitivity. AB - Adipocyte-specific secreted molecules, termed adipokines, have dispelled the notion of adipose tissue as an inert storage depot for lipids, and highlighted its role as an active endocrine organ that monitors and alters whole-body metabolism and maintains energy homeostasis. One of these adipokines, adiponectin (also known as Acrp30, AdipoQ, and GBP28), has gained significant attention recently as a mediator of insulin sensitivity. Many clinical reports and genetic studies over the past few years demonstrate decreased circulating levels of this hormone in metabolic dysfunction, such as obesity and insulin resistance, in both humans and animal models. Pharmacologic adiponectin treatments in rodents increase insulin sensitivity, although the primary site and detailed mechanism of action is yet to be determined. The phenotypes of adiponectin-deficient and transgenic adiponectin-overproducing animal models underscore the role of adiponectin in the maintenance of glucose and lipid homeostasis. PMID- 12762968 TI - Fat-induced liver insulin resistance. AB - In vitro studies have established that free fatty acids (FFAs) are important regulators of hepatic glucose metabolism. FFAs can increase hepatic glucose release by increasing the amount and activity of glucose-6-phosphatase and multiple gluconeogenic enzymes. Elevated FFAs can also potentially decrease hepatic glucose uptake by decreasing hepatic glucokinase activity. In vivo studies in both animals and humans have shown a close correlation between changes in plasma FFAs and endogenous glucose production (EGP). Intervention studies have established that changes in plasma FFAs are accompanied by changes in the relative contribution of gluconeogenesis and glycogenolysis to EGP. The effects of a change in FFAs on EGP itself are more evident when compensatory changes in insulin secretion are prevented or when insulin secretion is impaired (eg, diabetes mellitus). The effects of elevated FFAs on splanchnic glucose uptake are less clear, in that they appear to have no effect in nondiabetic humans and may impair uptake in people with type 2 diabetes. PMID- 12762969 TI - The Diabetes Prevention Program. PMID- 12762971 TI - Vascular compliance in diabetes. AB - The measurement of vascular compliance has assumed increasing importance as a powerful predictor of cardiovascular and all-cause mortality. Arterial stiffness increases with the duration of diabetes, older age, and concomitant hypertension. Hyperglycemia may increase arterial stiffness in diabetes by reducing the bioactivity of endothelium-derived nitric oxide (NO) either by decreasing NO production or inactivating NO by interaction with oxygen-derived free radicals. New approaches to therapy, such as the use of advanced glycation end product "breakers," may potentially benefit patients with diabetes. PMID- 12762973 TI - Managing lipid disorders: further evaluation of the VA-HIT. PMID- 12762970 TI - Heart disease in diabetic patients. AB - Both type 1 and type 2 diabetic patients have an increased incidence of ischemic heart disease and congestive heart failure. Cardiovascular disease accounts for up to 80% of the excess mortality in patients with type 2 diabetes. The burden of cardiovascular disease is especially pronounced in diabetic women. Factors that underlie diabetic heart disease include multiple vessel coronary artery disease, long-standing hypertension, metabolic derangements such as hyperglycemia and dyslipidemia, microvascular disease, and autonomic neuropathy. There is also increased sudden death associated with diabetes, which is due, in part, to the underlying autonomic neuropathy. This article reviews diabetic cardiac disease, with an emphasis on type 2 diabetes. PMID- 12762972 TI - Prevention of type 2 diabetes. AB - Diabetes is a major public health problem that is approaching epidemic proportions in our society and worldwide. Cardiovascular disease is the major cause of morbidity and mortality in people with diabetes. Control of cardiovascular disease risk factors is achieved only in a minority of patients. Given the magnitude of the problem and the seriousness of diabetes complications, prevention appears to be a logical approach to curb the rising prevalence of the disease. Interventions such as lifestyle modifications and the use of metformin and acarbose have been shown in randomized prospective trials to prevent diabetes in high-risk patients. Other interventions are currently being examined in large prospective studies. It is likely that one or a combination of these approaches will make diabetes prevention a reality in the near future. PMID- 12762974 TI - Inflammation and emerging risk factors in diabetes mellitus and atherosclerosis. AB - Type 2 diabetes mellitus and atherosclerosis are complex and progressive conditions that share several common antecedents. Recent data suggest that inflammation may play a central role in the origins and complications of cardiovascular disease and, possibly, type 2 diabetes mellitus. C-reactive protein and plasminogen activator inhibitor-1 are circulating markers of low grade inflammation, thrombosis, and vascular injury. Together with homocysteine, they have been associated with the underlying inflammatory processes and are considered to be "nontraditional" risk factors of atherosclerosis. The role of their measurement in clinical practice remains unclear. In this article, we review the available evidence demonstrating a link between inflammation, cardiovascular disease, and diabetes. We discuss how therapeutic agents used for both cardiovascular disease and diabetes modulate the inflammatory responses and possibly attenuate the complications of these two chronic disorders that cause significant morbidity and mortality. PMID- 12762975 TI - Type 2 diabetes mellitus in adolescence: lipid and cardiovascular risk factors. AB - This review discusses the important consequences of dyslipidemia and arteriosclerosis in type 2 diabetes as documented in studies in adults. It then examines the relatively recent upsurge in type 2 diabetes in children and adolescents, its characteristics, and its importance in directing our attention to cardiovascular risk factors in this age group. The discussion concludes with an examination of the information available about arteriosclerosis in the young and about the treatment of hypercholesterolemia in children and adolescents. PMID- 12762980 TI - Heteronuclear spin decoupling in solid-state NMR under magic-angle sample spinning. AB - Achieving high spectral resolution is an important prerequisite for the application of solid-state NMR to biological molecules. Higher spectral resolution allows to resolve a larger number of resonances and leads to higher sensitivity. Among other things, heteronuclear spin decoupling is one of the important factors which determine the resolution of a spectrum. The process of heteronuclear spin decoupling under magic-angle sample spinning is analyzed in detail. Continuous-wave RF irradiation leads only in a zeroth-order approximation to a full decoupling of heteronuclear spin systems in solids under magic-angle spinning (MAS). In a higher-order approximation, a cross-term between the dipolar coupling tensor and the chemical-shielding tensor is reintroduced, providing a scaled coupling term between the heteronuclear spins. In strongly coupled spin systems this second-order recoupling term is partially averaged out by the proton spin-diffusion process, which leads to exchange-type narrowing of the line by proton spin flips. This process can be described by a spin-diffusion type superoperator, allowing the efficient simulation of strongly coupled spin systems under heteronuclear spin decoupling. Low-power continuous-wave decoupling at fast MAS frequencies offers an alternative to high-power irradiation by reversing the order of the averaging processes. At fast MAS frequencies low-power continuous wave decoupling leads to significantly narrower lines than high-power continuous wave decoupling while at the same time reducing the power dissipated in the sample by several orders of magnitude. The best decoupling is achieved by multiple-pulse sequences at high RF fields and under fast MAS. Two such sequences, two-pulse phase-modulated decoupling (TPPM) and X-inverse-X decoupling (XiX), are discussed and their properties analyzed and compared. PMID- 12762981 TI - Stochastic excitation and Hadamard correlation spectroscopy with bandwidth extension in RF FT-EPR. AB - The application of correlation spectroscopy employing stochastic excitation and the Hadamard transform to time-domain Fourier transform electron paramagnetic resonance (FT-EPR) spectroscopy in the radiofrequency (RF) band is described. An existing, time-domain FT-EPR spectrometer system with a Larmor frequency (L(f)) of 300 MHz was used to develop this technique by incorporating a pseudo-random pulse sequence generator to output the maximum length binary sequence (MLBS, 10- and 11-bit). Software developed to control the EPR system setup, acquire the signals, and post process the data, is outlined. The software incorporates the Hadamard transform algorithm to perform the required cross-correlation of the acquired signal and the MLBS after stochastic excitation. To accommodate the EPR signals, bandwidth extension was accomplished by sampling at a rate many times faster than the RF pulse repetition rate, and subsequent digital signal processing of the data. The results of these experiments showed that there was a decrease in the total acquisition time, and an improved free induction decay (FID) signal-to-noise (S/N) ratio compared to the conventional coherent averaging approach. These techniques have the potential to reduce the RF pulse power to the levels used in continuous wave (CW) EPR while retaining the advantage of time domain EPR methods. These methods have the potential to facilitate the progression to in vivo FT-EPR imaging of larger volumes. PMID- 12762976 TI - Combination lipid-lowering therapy in diabetes. AB - Owing to the National Cholesterol Education Program Adult Treatment Panel III recommendations that patients with diabetes require a low-density lipoprotein (LDL) less than 100 mg/dL and a non-high-density lipoprotein (HDL) less than 130 mg/dL, frequently, combination lipid-lowering therapy is required. However, diabetic patients are commonly on multiple medications and have renal impairment. Therefore, the risk of myopathy with statin therapy is markedly increased. The safety of lipid-lowering therapy can be significantly improved by avoiding high dose statins in combination with fibrates, especially gemfibrozil. To achieve non HDL goals combining fenofibrate, or if glucose is well controlled, niacin, with a statin (not to exceed 40 mg), may significantly reduce the risk of myopathy. For diabetic patients who require additional LDL lowering, ezetimibe may provide a safe combination to a statin to achieve the LDL goal of less than 100 mg/dL. PMID- 12762982 TI - Phase-modulated heteronuclear decoupling in NMR of solids. AB - A theoretical description of heteronuclear decoupling in solids, which takes into account homonuclear dipole-dipole interactions and spinning of the sample, is presented. Based on this analysis, a simple and efficient decoupling sequence nPPM is introduced. It consists of n/4 TPPM subcycles, followed by another n/4 subcycles with reversed phases. PMID- 12762983 TI - Band-selective recoupling of homonuclear double-quantum dipolar interaction with a generalized composite 0 degrees pulse: application to 13C aliphatic region selective magnetization transfer in solids. AB - Recoupling of homonuclear double quantum (DQ)-dipolar interactions is a useful technique for the structural analysis of molecules in solids. We have designed a series of elemental 0 degrees pulses for the recoupling sequences with the rf phase rotation about the z-axis, known as CN. The proposed 0 degrees pulses whose total flip angle >/=360 degrees provide spin rotation vectors in the xy-plane. Thus, the residual spin rotation can be canceled by rf phase rotation about the z axis. An analysis by the coherent averaging theory showed that effective bandwidths of the recoupling sequences are limited not by the reduction in the dipolar scaling factor but by the increase in the residual spin rotation due to offset. A CN sequence with these elemental pulses provides an effective bandwidth of DQ-dipolar recoupling from ca. 0.5nu(R) to 4nu(R) for numerical simulations. Here, nu(R) is the sample spinning frequency. The 0 degrees pulses were applied to band-selective recoupling for the magnetization transfer in uniformly 13C labeled molecules. Narrow-band recoupling enhances the magnetization transfer between spins within the effective range by decoupling the dipolar interactions between spins one of which is outside the range. The narrow band operation reduces rf field strength, which improves the CH decoupling. Increases in signal intensities by the use of the proposed 0 degrees pulses are experimentally shown for 13C-labeled amino acids. PMID- 12762984 TI - Difference-NMR techniques for selection of components on the basis of relaxation times. AB - This work describes a numerical methodology to obtain more efficient relaxation filters to selectively retain or remove components based on relaxation times. The procedure uses linear combinations of spectra with various recycle or filter delays to obtain components that are both quantitative and pure. Modulation profiles are calculated assuming exponential relaxation behavior. The method is general and can be applied to a wide range of solution or solid-state NMR experiments including direct-polarization (DP), or filtered cross-polarization (CP) spectra. 13C NMR experiments on isotactic poly(1-butene) and dimethyl sulfone showed the utility of the technique for selectively suppressing peaks. PMID- 12762986 TI - Carbon-13 lineshapes in solid-state NMR of labeled compounds. Effects of coherent CSA-dipolar cross-correlation. AB - The experimental lineshapes of the carboxyl and methyl carbon resonances of fully 13C enriched L-Alanine are studied in detail at different MAS frequencies and decoupling field strengths. Complex lineshapes at intermediate spinning speeds were explained by the joint effect of off rotational resonance and coherent CSA dipolar cross-correlation. Whereas off rotational-resonance effects lead to complex lineshapes due to a splitting of some energy levels, coherent CSA-dipolar cross-correlation introduces either a differential intensity and/or a differential broadening of the lines of the J-multiplet. The conditions which lead to such effects are explained and experimentally verified. Additional simulations show that these effects can be expected over a wide range of static magnetic fields and are not restricted to L-Alanine. PMID- 12762985 TI - Progress on the two-dimensional filter diagonalization method. An efficient doubling scheme for two-dimensional constant-time NMR. AB - An efficient way to treat two-dimensional (2D) constant-time (CT) NMR data using the filter diagonalization method (FDM) is presented. In this scheme a pair of N- and P-type data sets from a 2D CT NMR experiment are processed jointly by FDM as a single data set, twice as large, in which the signal effectively evolves in time for twice as long. This scheme is related to "mirror-image" linear prediction, but with the distinction that the data are directly used, without any preprocessing such as Fourier transformation along one dimension, or point-by point reflection. As the signal has nearly perfect Lorentzian line shape in the CT dimension, it can be efficiently handled by the FDM approach. Applied to model and experimental signals, the scheme shows significant resolution improvement, and appears to tolerate noise reasonably well. Other complex aspects of multidimensional FDM are discussed and illustrated. PMID- 12762987 TI - Complexation-induced chemical shifts--ab initio parameterization of transferable bond anisotropies. AB - Complexation-induced changes in proton chemical shifts provide a potent tool for conformational analysis, being highly dependent on intermolecular orientation. An important contribution to these shifts arises from the molecular magnetisability anisotropy, or more specifically from the anisotropy of certain groups, such as aromatic rings and unsaturated bonds. While the influence of aromatic rings has been well characterised via the ring current effect, unsaturated bonds have received much less attention and prediction of complexation shifts is hampered by the lack of accurate anisotropy parameters for these bonds. We have therefore used ab initio calculations at the HF/aug-cc-pVDZ level to obtain bond anisotropies for C-H, N-H, C=O, C=C, C triple bond N, N=N, C triple bond C, and C triple bond N. Fitting the anisotropies to bond magnetic dipoles (the McConnell equation) gives non-transferable values for C-H and N-H bonds. We have therefore expanded in terms of bond magnetic dipoles, quadrupoles, and octopoles for double and triple bonds only, obtaining highly accurate shielding surfaces in all cases. The transferable nature of the anisotropies is confirmed by comparing with shifts obtained in larger molecules containing unsaturated bonds. PMID- 12762988 TI - Artifacts in T(1rho)-weighted imaging: correction with a self-compensating spin locking pulse. AB - Significant artifacts arise in T(1rho)-weighted imaging when nutation angles suffer small deviations from their expected values. These artifacts vary with spin-locking time and amplitude, severely limiting attempts to perform quantitative imaging or measurement of T(1rho) relaxation times. A theoretical model explaining the origin of these artifacts is presented in the context of a T(1rho)-prepared fast spin-echo imaging sequence. Experimentally obtained artifacts are compared to those predicted by theory and related to B(1) inhomogeneity. Finally, a "self-compensating" spin-locking preparatory pulse cluster is presented, in which the second half of the spin-locking pulse is phase shifted by 180 degrees. Use of this pulse sequence maintains relatively uniform signal intensity despite large variations in flip angle, greatly reducing artifacts in T(1rho)-weighted imaging. PMID- 12762989 TI - In vivo NMR of hyperpolarized 3He in the human lung at very low magnetic fields. AB - We present NMR measurements of the diffusion of hyperpolarized 3He in the human lung performed at fields much lower than those of conventional MRI scanners. The measurements were made on standing subjects using homebuilt apparatus operating at 3mT. O(2)-limited transverse relaxation (T(2) up to 15-35s) could be measured in vivo. Accurate global diffusion measurements have been performed in vivo and in a plastic bag; the average apparent diffusion coefficient (ADC) in vivo was 14.2+/-0.6mm(2)/s, whereas the diffusion coefficient in the bag (3He diluted in N(2)) was 79.5+/-1mm(2)/s. 1D ADC mapping with high SNR ( approximately 200-300) demonstrates the real possibility of performing quality lung imaging at extremely low fields. PMID- 12762990 TI - Spectral restoration from low signal-to-noise, distorted NMR signals: application to hyphenated capillary electrophoresis-NMR. AB - In capillary electrophoresis separations coupled to NMR signal detection using small solenoidal coils, electrophoretic currents cause substantial distortion in the NMR spectral linewidths and peak heights, distortions which cannot be fully counteracted through shimming. The NMR spectra also have a low signal-to-noise ratio due to the small amounts of material, typically <1nmol, associated with such microseparations. This study proposes a two-step, signal processing method to restore spectral lines from the distorted NMR spectrum. First, a reference signal is acquired to estimate the broadening function, as a combination of several Lorentzian functions, using a gradient descent method. Then multi resolution wavelet analysis is applied to the distorted spectrum to determine an initial estimate of the frequencies of the spectral lines. Convergence to the final spectrum, a second set of Lorentzians, involves deconvolution with the estimated broadening function using a gradient descent method. Experimental CE NMR data show that considerable improvements in spectral quality are possible using this approach, although fine splittings can not be resolved if the broadening function is large. PMID- 12762991 TI - Inverse methods in two-dimensional NMR spectral analysis. AB - Solid-state NMR is a valuable technique for the study of disordered materials. Analysis of such spectra usually involves solution of so-called ill-posed inverse problems. Here we present a strategy for the analysis of two-parameter two dimensional NMR problems and test it on 2D DECODER and DOQSY experiments. Using Monte Carlo tests, constraints are determined for the resolution and accuracy of the analysis for both experiments. The methods are finally applied to spectra of spider dragline silk, a heterogeneous solid fibrous protein. PMID- 12762992 TI - Frequency-domain Hadamard spectroscopy. AB - A new technique is proposed for multichannel excitation and detection of NMR signals in the frequency domain, an alternative to the widely used pulse-excited Fourier transform method. An extensive array of N radiofrequency irradiation channels covers the spectrum of interest. A selective radiofrequency pulse sequence is applied to each channel, generating a steady-state NMR response acquired one-point-at-a-time in the intervals between pulses. The excitation pattern is repeated N times, phase-encoded according to a Hadamard matrix, and the corresponding N composite responses are decoded by reference to the same matrix. This multiplex technique offers the same sensitivity advantage as conventional Fourier transform spectroscopy. The irradiation pattern may be tailored to concentrate on interesting spectral regions, to facilitate homonuclear double resonance, or to avoid exciting strong solvent peaks. As no free induction decay is involved, the new method avoids problems of pulse breakthrough or lineshape distortion by premature termination of the time-domain signal. PMID- 12762993 TI - Anisotropy of collagen fiber orientation in sheep tendon by 1H double-quantum filtered NMR signals. AB - The anisotropy of the angular distribution of collagen fibrils in a sheep tendon was investigated by 1H double-quantum (DQ) filtered NMR signals. Double-quantum build-up curves generated by the five-pulse sequence were measured for different angles between the direction of the static magnetic field and the axis of the tendon plug. Proton residual dipolar couplings determined from the DQ build-up curves in the initial excitation/reconversion time regime which mainly represent the bound water are interpreted in terms of a model of spin-1/2 pairs with their internuclear axes oriented on average along the fibril direction in the presence of proton exchange. The angular distribution of collagen fibrils around the symmetry axis of the tendon measured by the anisotropy of the residual dipolar couplings was described by a Gaussian function with a standard deviation of 12 degrees +/-1 degrees and with the center of the distribution at 4 degrees +/-1 degrees. The existence of this distribution is directly reflected in the finite value of the residual dipolar couplings at the magic angle, the value of the angular contrast, and the oscillatory behavior of the DQ build-up curves. The 1H residual dipolar couplings were also measured from the doublets recorded by the DQ-filtered signals. From the angular dependence of the normalized splitting the angular distribution of the collagen fibrils was evaluated using a Gaussian function with a standard deviation of 19 degrees +/-1 degrees and with the center of distribution at 2 degrees +/-1 degrees. The advantages and disadvantages of these approaches are discussed. PMID- 12762994 TI - A statistical analysis of NMR spectrometer noise. AB - Estimation of NMR spectral parameters, using e.g. maximum likelihood methods, is commonly based on the assumption of white complex Gaussian noise in the signal obtained by quadrature detection. Here we present a statistical analysis with the purpose of discussing and testing the validity of this fundamental assumption. Theoretical expressions are derived for the correlation structure of the noise under various conditions, showing that in general the noise in the sampled signal is not strictly white, even if the thermal noise in the receiver steps prior to digitisation can be characterised as white Gaussian noise. It is shown that the noise correlation properties depend on the ratio between the sampling frequency and the filter cut-off frequency, as well as the filter characteristics. The theoretical analysis identifies conditions that are expected to yield non-white noise in the sampled signal. Extensive statistical characterisation of experimental noise confirms the theoretical predictions. The statistical methods outlined here are also useful for residual analysis in connection with validation of the model and the parameter estimates. PMID- 12762995 TI - An NMR technique for measurement of magnetic field gradient waveforms. AB - Almost all NMR imaging and localized spectroscopic methods fundamentally rely on the use of magnetic field gradients. It follows that precise information on gradient waveform shape and rise-times is often most useful in experimental MRI. We present a very simple and robust method for measuring the time evolution of a magnetic field gradient. The method is based on the analysis of the NMR signal in the time domain, and requires no specialized field measurement probes for its implementation. The technique makes use of the principal that for small flip angles the excitation profile is a good approximation to the Fourier transform of the radio frequency pulse shape. Creation of the NMR signal can be considered as an inverse Fourier transform and thus variation of the gradient strength during the excitation pulse influences the shape of the NMR signal. Although originally designed for measurement of the rise time only, we have now extended the technique to measure the exact time course of the gradient. The theory is confirmed by experimental results for gradient waveform field measurements in a high-field vertical bore system. PMID- 12762996 TI - Self- and mutual-diffusion coefficients measurements by 31P NMR 1D profiling and PFG-SE in dextran gels. AB - 31P NMR 1D profiling was successfully introduced to measure macroscale mutual diffusion coefficients (D(m)) of phosphate ions in dextran gels. Series of 1D profiles describing the phosphate concentration along cylindrical dextran gels were acquired at different times. These profiles that included over 600 points could be fitted using equations derived from Fick's law, with D(m) as the single fitting parameter. Release and penetration profiles were recorded providing two alternative approaches for allowing the determination of D(m). The D(m) values were compared with microscale self-diffusion coefficients (D(s)) measured by pulsed field gradient spin echo (PFG-SE) technique. D(m) values, measured between 25 and 45 degrees C, were systematically lower than D(s). The experimental diffusion time and the associated diffusion length of D(s) (60 ms, 10 microm) are short compared to those of D(m) (up to 18 h, 50 mm). These scale differences are considered to be the origin of different D(s) and D(m) and provide information relative to the network in these gels. PMID- 12762997 TI - The DIVAM sequence: selective excitation of signals from both rigid and mobile domains in a fluoropolymer. AB - Simulations have been carried out on z-magnetizations produced by (and hence of the resulting spectra from) the dipolar filter (DF) and the recently suggested "Discrimination Induced by Variable-Amplitude Minipulses" (DIVAM) pulse sequences [S. Ando et al., Polymer 42 (2001) 8137; Magn. Reson. Chem. 40 (2002) 97]. The strengths of dipolar interactions have been modelled by introducing different values for transverse relaxation times. The DF case has been extended by allowing the pulse angles to be smaller than 90 degrees. The pulse intervals have also been used as variables. For the DIVAM case, the variables are similarly the minipulse nutation angles and minipulse intervals. The computations show that DIVAM is superior to DF in terms of selectivity for spectra of heterogeneous materials such as semi-crystalline polymers. The effects of the pulse sequences on 19F spectra of poly(vinylidene fluoride) (PVDF) and of a copolymer of vinylidene fluoride and trifluoroethylene (p(VDF/TrFE)) are presented, together with fits of the experimental results by the simulations. PMID- 12762998 TI - Recoupled long-range C-H dipolar dephasing in solid-state NMR, and its use for spectral selection of fused aromatic rings. AB - This work introduces a simple new solid-state 13C NMR method for distinguishing various types of aromatic residues, e.g. those of lignin from fused rings of charcoal. It is based on long-range dipolar dephasing, which is achieved by recoupling of long-range C-H dipolar interactions, using two 1H 180 degrees pulses per rotation period. This speeds up dephasing of unprotonated carbon signals approximately threefold compared to standard dipolar dephasing without recoupling and thus provides much more efficient differential dephasing. It also reduces the effects of spinning-speed dependent effective proton-proton dipolar couplings on the heteronuclear dephasing. Signals of unprotonated carbons with two or more protons at a two-bond distance dephase to <3% within less than 0.9 ms, significantly faster than those of aromatic sites separated from the nearest proton by three or more bonds. Differential dephasing among different unprotonated carbons is demonstrated in a substituted anthraquinone and 3-methoxy benzamide. The data yield a calibration curve for converting the dephasing rates into estimated distances from the carbon to the nearest protons. This can be used for peak assignment in heavily substituted or fused aromatic molecules. Compared to lignin, slow dephasing is observed for the aromatic carbons in wood charcoal, and even slower for inorganic carbonate. Direct 13C polarization is used on these structurally complex samples to prevent loss of the signals of interest, which by design originate from carbons that are distant from protons and therefore crosspolarize poorly. In natural organic matter such as humic acids, this combination of recoupled dipolar dephasing and direct polarization at 7-kHz MAS enables selective observation of signals from fused rings that are characteristic of charcoal. PMID- 12762999 TI - Mechanisms underlying episodic gonadotropin-releasing hormone secretion. AB - The episodic secretion of gonadotropin-releasing hormone (GnRH) is crucial for fertility, but the cellular mechanisms and network properties generating GnRH pulses are not well understood. We will explore three primary aspects of this intermittent hormonal signal: the source of rhythm(s), the possible mechanisms comprising oscillator(s), and how GnRH neurons are synchronized to produce a pulse of hormone release into the pituitary portal blood. Current knowledge will be reviewed, and hypotheses and working models proposed for future studies. PMID- 12763000 TI - Molecular defects in the pathogenesis of pituitary tumours. AB - The majority of pituitary adenomas are trophically stable and change relatively little in size over many years. A comparatively small proportion behave more aggressively and come to clinical attention through inappropriate hormone secretion or adverse effects on surrounding structures. True malignant behaviour with metastatic spread is very atypical. Pituitary adenomas that come to surgery are predominantly monoclonal in origin and roughly half are aneuploid, indicating either ongoing genetic instability or transition through a period of genetic instability at some time during their development. Few are associated with the classical mechanisms of tumour formation but it is generally believed that the majority harbour quantitative if not qualitative differences in molecular composition compared to the normal pituitary. Despite their prevalence and the ready availability of biopsy material, at the present time, the precise molecular pathogenesis of the majority of pituitary adenomas remains unclear. This review summarizes current thinking. PMID- 12763001 TI - Molecular determinants and physiological relevance of extrasomatic RNA localization in neurons. AB - Specific sorting of mRNA molecules to subcellular microdomains is an evolutionarily conserved mechanism by which the polarized nature of eukayotic cells may be established and maintained. The molecular composition of the RNA localization machinery is complex. Sequence motifs within RNA molecules to be transported, called cis-acting elements, and proteins, referred to as trans acting factors, are essential components. Transport of the resulting ribonucleoprotein complexes to distinct cytoplasmic regions occurs along the cytoskeletal network. The pathway is observed in organisms as diverse as yeast and human and it plays a critical role in development and cell differentiation. Moreover, RNA localization takes place in differentiated cell types including neurons. There is ample evidence to suggest that sorting of defined mRNA species to the neurites of nerve cells and on-site translation has an impact on various aspects of nerve cell biology. PMID- 12763002 TI - Dynamic variations in affective priming. AB - The present study investigates the dynamics of emotional processing and awareness using an affective facial priming paradigm in conjunction with a multimodal assessment of awareness. Key facial primes are visually masked, and are presented for brief (unconscious) and extended (conscious) durations. Using a preference measure, we examine whether the effects of the primes differ qualitatively (Murphy & Zajonc, 1993). We show that: (a) unconscious affective priming with faces emerges strongly in initial presentations and diminishes rapidly with repetition; (b) conscious affective priming also emerges strongly in initial presentations, however it persists in strength with repetition; and (c) in contrast to other reports on the salience of negative stimuli, happy faces appear more salient than sad faces when presented outside awareness. We discuss the limits and extensions of unconscious affective priming with faces, and consider several methodological and conceptual questions concerning emotional processing out of awareness. PMID- 12763003 TI - Task unrelated thought: the role of distributed processing. AB - Task unrelated thought (TUT) refers to thought directed away from the current situation; for example, a day dream. Encapsulated models of cognition propose that qualitative changes in consciousness, i.e., the production of TUT, can be explained in terms of changes in the quantity of resources deployed for task completion. In contrast, distributed models of cognition emphasize the importance of holistic processes in the generation and maintenance of task focus and are consistent with the effects of higher order variables such as schemata. Three experiments were conducted on healthy participants using a categorical stimulus organization to contrast distributed and encapsulated views of cognition. Experiments 1 and 2 showed that despite the increased difficulty of the alphabetical verbal fluency task fewer TUTs were produced during category fluency condition. Experiment 3 replicated the categorical suppression of TUT while encoding and recalling information in a memory task. The results of these three experiments support the predictions derived from models of cognition emphasizing the importance of stimulus organization in the generation and maintenance of task focus and have potential importance for the scientific evaluation of cognition. PMID- 12763004 TI - Impaired strategic regulation of contents of conscious awareness in schizophrenia. AB - Conscious awareness comprises two distinct states, autonoetic and noetic awareness. Schizophrenia impairs autonoetic, but not noetic, awareness. We investigated the strategic regulation of relevant and irrelevant contents of conscious awareness in schizophrenia using a directed forgetting paradigm. Twenty one patients with schizophrenia and 21 normal controls were presented with words and told to learn some of them and forget others. In a subsequent test, they were asked to recognize all the words they had seen previously and give remember, know or guess responses according to whether they recognized words on the basis of autonoetic awareness, noetic awareness, or guessing. Overall, patients showed the same degree of a directed forgetting effect as normal subjects. However, whereas the effect was observed both for remember and know responses in normal subjects, it was observed for know, but not for remember, responses in patients. These results indicate that patients with schizophrenia exhibit an impaired strategic regulation of contents of autonetic awareness for relevant and irrelevant information. PMID- 12763005 TI - Does opposition logic provide evidence for conscious and unconscious processes in artificial grammar learning? AB - The question of whether studies of human learning provide evidence for distinct conscious and unconscious influences remains as controversial today as ever. Much of this controversy arises from the use of the logic of dissociation. The controversy has prompted the use of an alternative approach that places conscious and unconscious influences on memory retrieval in opposition. Here we ask whether evidence acquired via the logic of opposition requires a dual-process account or whether it can be accommodated within a single similarity-based account. We report simulations using a simple neural network model of two artificial grammar learning experiments reported by that dissociated conscious and unconscious influences on classification. The simulations demonstrate that opposition logic is insufficient to distinguish between single- and multiple-system models. PMID- 12763006 TI - Seeing is believing: the reality of hypnotic hallucinations. AB - Two experiments investigated the reality attributed to hypnotic suggestion through subtle projection of a visual image during simultaneous suggestion for a visual hallucination that resembled the projected image. In Experiment 1, high and low hypnotizable participants were administered either a hypnotic induction or wake instructions, given a suggestion to hallucinate a shape, and then the projected image was subsequently introduced. Although highs in both conditions rated the projected image more vividly than lows, highs in the hypnosis (but not wake) condition made comparable reality ratings when the projected image was absent and present. In Experiment 2, high hypnotizable participants were administered a suggestion to see a shape on a wall. For half the participants the suggested image was projected on the wall and then removed, and for half the projection was initially absent and then introduced. Participants who had the projection absent and then present reported comparable reality and vividness ratings when the projection was absent and present. These findings indicate that elevated hypnotizability and hypnosis are associated with attributions of external reality to suggested experiences. PMID- 12763007 TI - Functional neuroanatomy of altered states of consciousness: the transient hypofrontality hypothesis. AB - It is the central hypothesis of this paper that the mental states commonly referred to as altered states of consciousness are principally due to transient prefrontal cortex deregulation. Supportive evidence from psychological and neuroscientific studies of dreaming, endurance running, meditation, daydreaming, hypnosis, and various drug-induced states is presented and integrated. It is proposed that transient hypofrontality is the unifying feature of all altered states and that the phenomenological uniqueness of each state is the result of the differential viability of various frontal circuits. Using an evolutionary approach, consciousness is conceptualized as hierarchically ordered cognitive function. Higher-order structures perform increasingly integrative functions and thus contribute more sophisticated content. Although this implies a holistic approach to consciousness, such a functional hierarchy localizes the most sophisticated layers of consciousness in the zenithal higher-order structure: the prefrontal cortex. The hallmark of altered states of consciousness is the subtle modification of behavioral and cognitive functions that are typically ascribed to the prefrontal cortex. The theoretical framework presented yields a number of testable hypotheses. PMID- 12763008 TI - Halo and devil effects demonstrate valenced-based influences on source-monitoring decisions. AB - Source attributions based on positive versus negative valence were examined in four experiments. The two sources were individuals who were depicted positively or negatively, and the content of their statements was similarly valenced. When valenced information about the sources was provided after learning the statements, test biases to attribute positive statements to the positive source and negative statements to the negative source were strongly present. Providing the same information prior to learning improved memory, but did not entirely eliminate test biases based on valence. Signal detection analysis suggests that these "halo effect" biases are criterion-based and not memory-based. Therefore, the results are more consistent with descriptions of source-monitoring processes that can benefit from familiarity-based partial information as opposed to descriptions in which source monitoring is primarily recollection-based. PMID- 12763009 TI - Perceptual acceleration of objects in stream: evidence from flash-lag displays. AB - An object in continuous motion is perceived ahead of the briefly flashed object, although the two images are physically aligned (Nijhawan, 1994), the phenomenon called flash-lag effect. Flash-lag effects have been found also with other continuously changing features such as color, pattern entropy, and brightness (Sheth, Nijhawan, & Shimojo, 2000) as well as with streamed pre- and post-target input without any change of the feature values of streaming items in feature space (Bachmann & Poder, 2001a. 2001b). We interpret all instances of the flash lag as a consequence of a more fundamental property of conscious perception in general: acceleration of the speed with which samples of perceptual information become represented in explicit format immediately after the stimulation onset. Decreased visual latency of the samples of stimulus information from the streamed input leads to the relative perceptual lag for the separately flashed stimulus because it is not preceded by adjacent sensory input that would have accelerated its perception. Experimental support for the notion of perceptual acceleration is reviewed. PMID- 12763010 TI - Continuity between waking activities and dream activities. AB - Empirical studies largely support the continuity hypothesis of dreaming. Despite of previous research efforts, the exact formulation of the continuity hypothesis remains vague. The present paper focuses on two aspects: (1) the differential incorporation rate of different waking-life activities and (2) the magnitude of which interindividual differences in waking-life activities are reflected in corresponding differences in dream content. Using a correlational design, a positive, non-zero correlation coefficient will support the continuity hypothesis. Although many researchers stress the importance of emotional involvement on the incorporation rate of waking-life experiences into dreams, formulated the hypothesis that highly focused cognitive processes such as reading, writing, etc. are rarely found in dreams due to the cholinergic activation of the brain during dreaming. The present findings based on dream diaries and the exact measurement of waking activities replicated two recent questionnaire studies. These findings indicate that it will be necessary to specify the continuity hypothesis more fully and include factors (e.g., type of waking-life experience, emotional involvement) which modulate the incorporation rate of waking-life experiences into dreams. Whether the cholinergic state of the brain during REM sleep or other alterations of brain physiology (e.g., down regulation of the dorsolateral prefrontal cortex) are the underlying factors of the rare occurrence of highly focused cognitive processes in dreaming remains an open question. Although continuity between waking life and dreaming has been demonstrated, i.e., interindividual differences in the amount of time spent with specific waking-life activities are reflected in dream content, methodological issues (averaging over a two-week period, small number of dreams) have limited the capacity for detecting substantial relationships in all areas. Nevertheless, it might be concluded that the continuity hypothesis in its present general form is not valid and should be elaborated and tested in a more specific way. PMID- 12763011 TI - Competition between automatic and controlled processes. AB - We investigated the competition between automatic and controlled processes in a word stem completion task. Prime-display duration and the prime-target interval were manipulated. On each trial a masked prime was displayed briefly, followed either immediately or after a delay by a word stem. The subjects were required to complete each stem with the first word that came to mind, to report any prime they could identify, and not to give as completion any identified prime. By the assumption that automatic processes require less stimulus input and can be completed faster than consciously controlled processes we expected a stronger performance contribution from automatic processes with the shorter prime-display durations and in the immediate stems condition. The results confirmed this expectation. The findings highlight that consciously controlled processes require more time to run their course than unconscious automatic processes. PMID- 12763012 TI - New functions of histamine found in histidine decarboxylase gene knockout mice. AB - Gene targeting techniques have revolutionized the investigation of the effects of bioactive substances in pathological and physiological conditions. Histamine synthesis is uniquely catalyzed by L-histidine decarboxylase. The knockout mice of this gene express no histamine-producing activity and lack histamine. These mice have been used to examine the mechanisms of histamine in several known phenotypes, e.g., gastric acid secretion, contraction of smooth muscles, vascular permeability, and awakening, and have also been used to explore unreported effects of histamine in the whole body. First, we will review the former mechanisms and then move to the latter, new effects. Especially, in the latter mechanisms, we focus on several important roles of histamine in angiogenesis, neutrophil and eosinophil recruitment, bacterial infection, and systemic anaphylaxis in this review. Moreover, to our surprise, the morphology of mast cells in the knockout mice was severely affected by the absence of histamine in terms of their granules. PMID- 12763014 TI - Lysophosphatidic acid stimulates CREB through mitogen- and stress-activated protein kinase-1. AB - Lysophosphatidic acid (LPA) is a growth factor-like phospholipid that elicits a variety of cellular responses in numerous cell types, including neurons, immune cells, and fibroblasts. In this report, we investigated the possibility that LPA activates the transcription factor cAMP response element-binding protein, CREB, in Rat-2 fibroblast cells. CREB is activated in many cells downstream of signaling events, such as growth factor and neurotrophin stimulation. We found that LPA rapidly stimulated phosphorylation of CREB at Ser133 in a time- and dose dependent manner, as revealed by immunoblot analysis with a phospho-specific antibody recognizing CREB on Ser133. LPA-induced phosphorylation of CREB was dependent on the activation of extracellular signal-regulated kinase 1/2 (ERK1/2) and p38 mitogen-activated protein kinase (p38 MAPK). Inhibition of ERK1/2 with PD98059 and of p38 MAPK with SB203580 efficiently blocked LPA-mediated phosphorylation of CREB. The LPA-induced CREB phosphorylation was abolished by H89, an inhibitor of mitogen- and stress-activated protein kinase-1 (MSK1). Together, these data suggest that LPA stimulates nuclear transcription factor CREB via mitogen-activated protein kinase signaling components, ERK1/2, p38 MAPK, and MSK1 in Rat-2 fibroblast cells. PMID- 12763013 TI - Isolation and expression of Napor/CUG-BP2 in embryo development. AB - The human neuroblastoma apoptosis-related RNA-binding protein NAPOR is an ELAV like RNA-binding protein with three characteristic RNA recognition motifs (RRMs). We report here the cloning and characterization of a zebrafish Napor that has a high sequence homology to human NAPOR protein. Whole-mount in situ hybridization analysis revealed that zebrafish napor is dynamically expressed in early development. In addition to its maternal expression, napor transcripts were detected in adaxial mesoderm cells and lateral neural plate cells at early somite stages. By 10-somite stage, napor expression was restricted to the central nervous system, having a specific expression domain of rhombomere 5 in the hindbrain. In 24 hpf embryo, napor was expressed in subsets of differentiating neural cells in the forebrain and hindbrain as well as somitic muscle cells. The number of napor-expressing neural cells was greatly increased in the mind bomb mutant that has neurogenic phenotype resulting from deficits in the Notch signaling pathway. Furthermore, overexpression of napor by RNA microinjection resulted in severe defects in nervous system and gastrulation, suggesting the need for tight control of napor gene regulation during embryo development. PMID- 12763015 TI - Interleukin-6 differentially regulates androgen receptor transactivation via PI3K Akt, STAT3, and MAPK, three distinct signal pathways in prostate cancer cells. AB - The effects of IL-6 on prostate cancer cells are well documented yet remain controversial. Some reports suggested that IL-6 could promote prostate cancer cell growth, while others showed that IL-6 could repress prostate cancer cell growth. Here, we systemically examined various IL-6 signaling pathways in prostate cancer cells and found that IL-6 could go through at least three distinct pathways to modulate the functions of androgen receptor (AR), a key transcriptional factor to control the prostate cancer growth. Our results show that IL-6 can enhance AR transactivation via either the STAT3 or MAPK pathways. In contrast, IL-6 can suppress AR transactivation via the PI3K-Akt pathway. Co existence of these various signaling pathways may result in either additive or conflicting effects on AR transactivation. Together, our results indicate that the balance of these various pathways may then determine the overall effect of IL 6 on AR transactivation. PMID- 12763016 TI - Transglucosylation of tertiary alcohols using cassava beta-glucosidase. AB - We have compared the ability of beta-glucosidases from cassava, Thai rosewood, and almond to synthesize alkyl glucosides by transglucosylating alkyl alcohols of chain length C(1)-C(8). Cassava linamarase shows greater ability to transfer glucose from p-nitrophenyl-beta-glucoside to secondary alcohol acceptors than other beta-glucosidases, and is unique in being able to synthesize C(4), C(5), and C(6) tertiary alkyl beta-glucosides with high yields of 94%, 82%, and 56%, respectively. Yields of alkyl glucosides could be optimized by selecting appropriate enzyme concentrations and incubation times. Cassava linamarase required pNP-glycosides as donors and could not use mono- or di-saccharides as sugar donors in alkyl glucoside synthesis. PMID- 12763017 TI - Expression of a soluble glycine binding domain of the NMDA receptor in Escherichia coli. AB - Glycine is an essential co-agonist of the excitatory N-methyl-D-aspartate (NMDA) receptor. The glycine binding site of this subtype of ionotropic glutamate receptors is formed by the S1 and S2 regions of the NR1 subunit. Here, different S1S2 fusion proteins were expressed and purified from Escherichia coli cultures, and refolding protocols were established allowing the production of 30 mg of soluble S1S2 fusion protein from 1 liter bacterial culture. After affinity purification and renaturation, two of the fusion proteins (S1S2 and S1S2-V1) bound the competitive glycine site antagonist [3H]MDL105,519 with K(d) values of 9.35 and 3.9 nM, respectively. In contrast, with three other constructs (S1S2M, S1S2-V2, and -V3) saturable ligand binding could not be obtained. These results redefine the S1S2 domains required for high-affinity glycine binding. Furthermore, our high-affinity binding proteins may be used for the large-scale production of the glycine binding core region for future structural studies. PMID- 12763019 TI - High-level expression of human glycosyltransferases in insect cells as biochemically active form. AB - cDNAs, encoding human beta1,4-galactosyltransferase (hGalT I, EC 2.4.1.22), human Galbeta1,3(4)-GlcNAc alpha2,3-sialyltransferase (hST3GalIII, EC 2.4.99), and human Galbeta1,4-GlcNAc alpha2,6-sialyltransferase (hST6Gal I, EC 2.4.99.1), were cloned from human cell lines. In order to express these glycosyltransferases as secreted form in insect cells, cDNAs were inserted into a novel baculovirus transfer vector equipped with the mouse IgM signal peptide and IgG binding domain of the Staphylococcus aureus protein A as an N-terminal fusion partner. About 14 mg hGalT I, 8 mg hST3GalIII, and 6.4 mg hST6Gal I were purified from 1 liter of recombinant baculovirus infected insect cell culture media. The specific activities of recombinant hGalT I and hST6Gal I were determined as 0.65 and 1.6 U/mg protein, respectively. These results indicated that the recombinant hGalT I and hST6Gal I retained enzyme activities at similar level to those of the authentic one although they were fused with the IgG binding domain at the N terminus. Taken together, the mouse IgM signal peptide and IgG binding domain of the protein A could be efficiently used as an N-terminus fusion partner for the over-expression of heterologous proteins in insect cells. PMID- 12763018 TI - Lysine hydroxylation of collagen in a fibroblast cell culture system. AB - The lysine (Lys) hydroxylation pattern of type I collagen produced by human fibroblasts in culture was analyzed and compared. Fibroblasts were cultured from normal human skin (NSF), keloid (KDF), fetal skin (FDF), and skin tissues of Ehlers-Danlos syndrome type VIA and VIB patients (EDS-VIA and -VIB). The type I collagen alpha chains with or without non-helical telopeptides were purified from the insoluble matrix and analyzed. In comparison with NSFs, KDF and FDF showed significantly higher Lys hydroxylation, particularly in the telopeptide domains of both alpha chains. Both EDS-VIA and -VIB showed markedly lower Lys hydroxylation in the helical domains of both alpha chains whereas that in the telopeptides was comparable with those of NSFs. A similar profile was observed in the tissue sample of the EDS-VIB patient. These results demonstrate that the Lys hydroxylation pattern is domain-specific within the collagen molecule and that this method is useful to characterize the cell phenotypes in normal/pathological connective tissues. PMID- 12763020 TI - Spatial and temporal regulation of mitogen-activated protein kinase phosphorylation in the mouse suprachiasmatic nucleus. AB - Circadian and photic regulation of mitogen-activated protein kinase (MAPK) has been shown to associate closely with the function of the circadian clock in vertebrate clock tissues such as the mouse suprachiasmatic nucleus (SCN). Here we show that, in the central region of the mouse SCN, MAPK exhibited circadian and daily rhythms in phosphorylation with a peak at (subjective) night, and this activation was sustained for at least 8 h. In contrast, in the dorsomedial region of the SCN, MAPK showed an overt rhythm in phosphorylation with a transient peak at early subjective day, which was antiphase to that in the central region. Noticeably, the phospho-MAPK-immunoreactive cells observed in the dorsomedial region were distributed from the rostral to the caudal end of the SCN, whereas those observed in the central region were localized within the middle SCN along the rostral-caudal axis. Furthermore, a 15-min light pulse given at subjective night transiently evoked MAPK phosphorylation throughout the ventrolateral region of the SCN peaking within 15 min after the light onset, whereas nighttime phosphorylated MAPK signals in the central-middle SCN become undetectable within 60 min after the light onset. Thus, the mode of circadian and photic regulation of MAPK phosphorylation varies remarkably among the three subregions within the SCN, suggesting divergent and cell type-specific roles of MAPK in the clock system of the mouse SCN. PMID- 12763021 TI - APH1, PEN2, and Nicastrin increase Abeta levels and gamma-secretase activity. AB - A major component of the amyloid plaque core in Alzheimer's disease (AD) is the 40-42-residue amyloid beta peptide (Abeta). Mutations linked to AD such as those in presenilins 1 (PS1) and 2 (PS2) invariably increase the longer Abeta42 species that forms neurotoxic oligomers. It is believed that PS1/2 constitute the catalytic subunit of the gamma-secretase responsible for the final step in Abeta biogenesis. Recent genetic studies have identified a number of additional genes encoding APH1a, APH1b, PEN2, and Nicastrin proteins, which are part of the gamma secretase complex with PS1. Further, knockout studies using RNAi showed that these components are essential for gamma-secretase activity. However, the nature of gamma-secretase and how the aforementioned proteins regulate its activity are still incompletely understood. Here we present evidence that unlike PS1, overexpression of these proteins can increase the levels of Abeta, suggesting that these proteins are limiting for gamma-secretase activity. In addition, our studies also suggest that the presenilin partners regulate the relative levels of Abeta40 and Abeta42. PMID- 12763022 TI - Oxidative stress impairs endocytosis of the scavenger receptor class A. AB - We report the characterization of a cell system employing Chinese hamster ovary (CHO) cells and CHO cells transfected with the scavenger receptor class A (CHO SRA) using extracellularly produced reactive oxygen species (ROS) in order to study the endocytic function of the scavenger receptor. The oxidative environment was produced using tert-butyl hydroperoxide (TBH) and characterized by flow cytometry and cell viability. Once an adequate oxidative environment was established, binding and internalization studies of radiolabeled acetylated LDL particles (125I-labeled Ac-LDL) with CHO-SRA cells were carried out. RT-PCR analysis using total RNAs from CHO-SRA cells revealed that oxidative stress does not alter the expression of the scavenger receptor. However, internalization of 125I-labeled Ac-LDL through this receptor carried out by these cells was completely abolished under extracellularly oxidative conditions. Together, these results support the idea that an oxidative stress produced extracellularly, inhibiting the endocytosis of the scavenger receptor, could help to understand and explain the mechanisms by which several physiologically important ligands are accumulated in the extracellular space with its consequent cell damage. PMID- 12763023 TI - The fate of the prion protein in the prion/plasminogen complex. AB - The cellular prion protein (PrP(c)) forms complexes with plasminogen. Here, we show that the PrP(c) in this complex is cleaved to yield fragments of PrP(c). The cleavage is accelerated by plasmin but does not appear to be dependent on it. PMID- 12763024 TI - Identification of Ncd tail domain-binding sites on the tubulin dimer. AB - The Drosophila non-claret disjunctional (Ncd) kinesin-like protein is required for spindle assembly in oocytes and spindle maintenance in early embryos. Through the action of ATP-dependent microtubule (MT)-binding sites in the head and ATP independent MT-binding sites in the tail, Ncd may bundle and, perhaps, slide MTs relative to each other. Our previous work on the MT-binding site of the Ncd tail domain demonstrated that this site, like the MT-binding sites of tau, contains basic residues flanked by proline residues and can promote MT assembly and stability. Here, we characterize the interactions of a monomeric Ncd tail protein with subtilisin-digested MTs in order to identify sites on the tubulin dimer that interact with the Ncd tail. The results provide evidence for four such binding sites per tubulin dimer and support the hypothesis that each binding site consists of a cluster of acidic residues in the C-terminal regions of alpha- and beta-tubulin. PMID- 12763026 TI - On-chip single-cell microcultivation assay for monitoring environmental effects on isolated cells. AB - We have developed a on-chip single-cell microcultivation assay as a means of observing the adaptation process of single bacterial cells during nutrient concentration changes. This assay enables the direct observation of single cells captured in microchambers made on thin glass slides and having semipermeable membrane lids, in which cells were kept isolated with optical tweezers. After changing a medium of 0.2% (w/v) glucose concentration to make it nutrient-free 0.9% NaCl medium, the growth of all cells inserted into the medium stopped within 20 min, irrespective of their cell cycles. When a nutrient-rich medium was restored, the cells started to grow again, even after the medium had remained nutrient-free for 42 h. The results indicate that the cell's growth and division are directly related to their nutrient condition. The growth curve also indicates that the cells keep their memory of what their growth and division had been before they stopped growing. PMID- 12763025 TI - Peptide-based, irreversible inhibitors of gamma-secretase activity. AB - The characterization of the enzymes responsible for amyloid beta-peptide (Abeta) production is considered to be a primary goal towards the development of future therapeutics for the treatment of Alzheimer's disease. Inhibitors of gamma secretase activity were critical in demonstrating that the presenilins (PSs) likely comprised at least part of the active site of the gamma-secretase enzyme complex, with two highly conserved membrane aspartates presumably acting as catalytic residues. However, whether or not these aspartates are actually the catalytic residues of the enzyme complex or are merely essential for normal PS function and/or maturation is still unknown. In this paper, we report the development of reactive inhibitors of gamma-secretase activity that are functionally irreversible. Since such inhibitors have been shown to bind catalytic residues in other aspartyl proteases (e.g., HIV protease), they might be used to determine if the transmembrane aspartates of PSs are involved directly in substrate cleavage. PMID- 12763028 TI - Efficient and specific down-regulation of prion protein expression by RNAi. AB - Prion diseases are fatal neurodegenerative disorders associated with an abnormal isoform of the PrPc host-encoded protein. Invalidation of the Prnp gene, that encodes PrPc, led to transgenic mice that are viable, apparently healthy, and resistant to challenge by the infectious agent. These results indicated that a down-regulation of the Prnp gene expression is a potential therapeutic approach. In the present report, we demonstrate that RNAi targeted towards the Prnp mRNA can efficiently and highly specifically reduce the level of PrPc in transfected cells. It, thus, indicates that RNAi is an attractive therapeutic approach to fight against prion diseases. PMID- 12763027 TI - Differential effects of quercetin and resveratrol on Band 3 tyrosine phosphorylation signalling of red blood cells. AB - The protective effects of eating fruits and vegetables in the prevention of several degenerative pathologies have been attributed at least in part to the antioxidant and anti-inflammatory properties of polyphenols. In this study, we investigated the effects of two polyphenols, quercetin and resveratrol, on red blood cell Band 3 tyrosine phosphorylation signalling activated by peroxynitrite. Peroxynitrite is a physiological oxidant scavenged largely by the erythrocyte and formed by the reaction between nitrogen monoxide and superoxide anion. Quercetin and its structurally analogous (+)-catechin inhibited the peroxynitrite-dependent upregulation of Band 3 tyrosine phosphorylation. Quercetin was found to downregulate the activity of syk, which is upstream in the Band 3 tyrosine phosphorylation cascade, and partially prevented peroxynitrite-mediated phosphotyrosine phosphatase inhibition. Resveratrol and hydroxytyrosol, unexpectedly, amplified peroxynitrite-dependent upregulation of Band 3 tyrosine phosphorylation through the activation of lyn, a kinase of the src family. The present results clearly indicate that polyphenols may activate cell transduction pathways in different and sometimes opposite ways. PMID- 12763029 TI - p38 MAP kinase negatively regulates angiotensin II-mediated effects on cell cycle molecules in human coronary smooth muscle cells. AB - Many of the signaling events in VSMC stimulated by angiotensin II (AngII) are mediated by members of the mitogen-activated protein kinase (MAPK) family, including p38 MAPK. The role of p38 MAPK in AngII-mediated cell cycle regulation is poorly understood. Therefore, we examined the involvement of p38 MAPK signaling in AngII-stimulated DNA synthesis, phosphorylation of the retinoblastoma protein (Rb), and expression of the G1-phase cyclin D1 in human coronary artery smooth muscle cells (CASMC). AngII (1 microM) stimulated p38 MAPK and ERK1/2 activation. Pretreatment with the p38 MAPK inhibitors SB203580 (10 microM) (SB) or SKF-86002 (10 microM) (SKF) potently inhibited AngII-induced p38 MAPK activation, but enhanced AngII-mediated ERK1/2 activation. AngII-induced phosphorylation of Rb (Ser 795 and Ser 807/811), -cyclin D1 expression, and -DNA synthesis was also markedly enhanced by pharmacological inhibition of the p38 MAPK pathway. The present study demonstrates that p38 MAPK negatively regulates AngII-induced ERK1/2 activity, Rb phosphorylation, cyclin D1 expression, and DNA synthesis in human CASMC. These findings support an important role for p38 MAPK in modulating AngII-mediated VSMC hyperplasia. PMID- 12763031 TI - Carbonylation of ER chaperone proteins in aged mouse liver. AB - Progressive accumulation of oxidative damage to macromolecules in aged tissues is thought to contribute to the decline in tissue function characteristic of the aged phenotype. Mitochondria are a major intracellular source of reactive oxygen species (ROS); however, other organelles are also endogenous sources of oxyradicals and oxidants, which can damage macromolecules. We, therefore, sought to examine the relationship between aging and oxidative damage to ER resident proteins, which exist in a strongly oxidizing environment necessary for disulfide bond formation. In these studies, we have fractionated young and aged liver homogenates, resolved the proteins by 2D gel electrophoresis, assayed for oxidative damage as indicated by protein carbonylation, and identified BiP/Grp78, protein disulfide isomerase (PDI), and calreticulin as exhibiting an age associated increase in oxidative damage. Increased carbonylation of these key proteins in aged liver suggests an age-associated impairment in protein folding, disulfide crosslinking, and glycosylation in the aged mouse liver. PMID- 12763030 TI - Hepatic scavenger receptor class B, type I is stimulated by peroxisome proliferator-activated receptor gamma and hepatocyte nuclear factor 4alpha. AB - Excessive cellular cholesterol is transported to the liver by a pathway called 'reverse cholesterol transport.' Scavenger receptor class B, type I (SR-BI) mediates cholesterol uptake in the liver. Polyunsaturated fatty acids, known to activate peroxisome proliferator-activated receptor (PPAR), have been reported to increase hepatic cholesterol uptake. We found in the present study that PPARgamma induces expression of SR-BI in rat hepatocytes, liver endothelial cells, and Kupffer cells. In contrast, PPARalpha increased SR-BI levels only in hepatocytes and liver endothelial cells. PPARgamma/RXR binds to a response element between 459 and -472 bp in the human SR-BI promoter. Furthermore, hepatocyte nuclear factor 4alpha (HNF4alpha) was found to enhance PPARgamma-mediated SR-BI transcription. Thiazolidinedione (TZD)-activated PPARgamma/RXR increased hepatic SR-BI levels, which may lead to increased hepatic cholesterol uptake and less accumulation of lipids in peripheral tissues. The present results are in agreement with previous reports, indicating that specific PPARgamma-agonists (such as TZDs) protect against atherosclerosis. PMID- 12763032 TI - Distinct pathways of extracellular signal-regulated kinase activation by growth factors, fibronectin and parathyroid hormone 1-34. AB - Growth factors, hormones, and matrix proteins regulate osteoblast proliferation and differentiation, acting through cognate receptors. Since each of the receptors are coupled to a variety of distinct signal transduction pathways, in this report we evaluated whether there is a common convergent intermediate step that allows cross-talk among the various pathways. Since extracellular signal regulated kinases 1 and 2 (Erk1/2) play a role in mitogenesis and differentiation processes, we evaluated the effects of various osteotrophic factors on Erk1/2 phosphorylation in osteoblasts. Osteoblasts isolated from the metaphyseal marrow (MM) and diaphyseal marrow (DM) of 4-6 week old male rat longitudinal bones were grown to confluency and Erk1/2-phosphorylation was evaluated using antibodies that recognized either the total or the phosphorylated form of the kinase. There was very little Erk1/2 phosphorylation in cells kept in suspension. Both MM and DM cells attached to fibronectin (FN), demonstrated Erk1/2 phosphorylation that persisted for at least up to 8h. Platelet-derived growth factor AB (PDGF-AB) induced a transient and robust Erk1/2 phosphorylation that was attenuated by 2h. Studies with specific inhibitors indicated that the effects of these factors were mediated by protein kinase C, by receptor tyrosine kinase, as well as by protein phosphatases. Parathyroid hormone (PTH 1-34), a bone anabolic agent however, caused a down-regulation of FN stimulated Erk1/2 phosphorylation in MM derived cells. The inhibitory effect of PTH was mediated through cAMP-dependent protein kinase A (PKA) activation. The data collectively suggest that a combination of diverse extracellular stimuli regulates Erk1/2 phosphorylation that may ultimately influence osteoblast proliferation and/or differentiation. PMID- 12763033 TI - Molecular characterisation and expression analysis of the first hemicellulase gene (bxl1) encoding beta-xylosidase from the thermophilic fungus Talaromyces emersonii. AB - The gene coding for beta-xylosidase, bxl1, has been cloned from the thermophilic filamentous fungus, Talaromyces emersonii. This is the first report of a hemicellulase gene from this novel source. At the genomic level, bxl1 consists of an open reading frame of 2388 nucleotides with no introns that encodes a putative protein of 796 amino acids. The bxl1 translation product contains a signal peptide of 21 amino acids that yields a mature protein of 775 amino acids, with a predicted molecular mass of 86.8 kDa. The deduced amino acid sequence of bxl1 exhibits considerable homology with the primary structures of the Aspergillus niger, Aspergillus nidulans, Aspergillus oryzae, and Trichoderma reesei beta xylosidase gene products, and with some beta-glucosidases, all of which have been classified as Family 3 glycosyl hydrolases. Northern blot analysis of the bxl1 gene indicates that it is induced by xylan and methyl-beta-D-xylopyranoside. D Xylose induced expression of bxl1 but was shown to repress induction of the gene at high concentrations. The presence of six CreA binding sites in the upstream regulatory sequence (URS) of the bxl1 gene indicates that the observed repression by D-glucose may be mediated, at least partly, by this catabolite repressor. PMID- 12763034 TI - Fusion-type lycopene beta-cyclase from a thermoacidophilic archaeon Sulfolobus solfataricus. AB - Examination of the sequence of a hypothetical gene with an unknown function included in the carotenogenic gene cluster in the genome of a thermoacidophilic archaeon Sulfolobus solfataricus led to the prediction that the gene encodes a novel-type lycopene beta-cyclase, whose N- and C-terminal halves are homologous to the subunits of the bacterial heterodimeric enzymes. The recombinant expression of the gene in lycopene-producing Escherichia coli resulted in the accumulation of beta-carotene in the cells, which verifies the function of the gene. Homologues of the archaeal lycopene beta-cyclase from various organisms such as bacteria, archaea, and fungi have been reported. Although their primary structures are clearly specific to the biological taxa, a phylogenetic analysis revealed the unexpected complicity of the evolutional route of these enzymes. PMID- 12763035 TI - Intracellular delivery of glutathione S-transferase into mammalian cells. AB - Protein transduction domains (PTDs) derived from human immunodeficiency virus Tat protein and herpes simplex virus VP22 protein are useful for the delivery of non membrane-permeating polar or large molecules into living cells. In the course of our study aiming at evaluating PTD, we unexpectedly found that the fluorescent dye-labeled glutathione S-transferase (GST) from Schistosoma japonicum without known PTDs was delivered into COS7 cells. The intracellular transduction of GST was also observed in HeLa, NIH3T3, and PC12 cells, as well as in hippocampal primary neurons, indicating that a wide range of cell types is permissive for GST transduction. Furthermore, we showed that the immunosuppressive peptide VIVIT fused with GST successfully inhibits NFAT activation. These results suggest that GST is a novel PTD which may be useful in the intracellular delivery of biologically active molecules, such as small-molecule drugs, bioactive peptides, or proteins. PMID- 12763036 TI - Dysregulated expression of MIC-1/PDF in human prostate tumor cells. AB - As a part of the study to identify genes associated with hormone-refractory stage of human prostate cancer, we have recently identified several genetic and epigenetic changes that seem to be associated with the progression of androgen sensitive to androgen-independent prostate tumor cells. In the present study, we report a novel gene, macrophage inhibitory cytokine-1 (MIC-1) also known as prostate derived factor (PDF), that was highly expressed in androgen-independent LNCaP-C81 cells and its metastatic variant LNCaP-Ln3 compared to androgen sensitive LNCaP-C33 cells. The MIC-1/PDF expression was dysregulated (very low to non-detectable) in the androgen-independent PC3 and DU145 cells. Interestingly, serum factors demonstrated a differential regulation of MIC-1/PDF in the androgen sensitive and the androgen-independent cells of LNCaP cells. Immunohistochemical analysis on 15 prostatic adenocarcinomas showed a weak staining in the benign prostatic glandular area (intensity score 2.38+/-0.25; n=13), while the immunoreactivity was significantly stronger (p<0.05) in areas of adenocarcinoma (score 7.33+/-0.88; n=15). Altogether, these data suggest that the serum factors (including androgens and cytokines) might contribute to the regulation of the MIC 1/PDF gene that seems to be associated with the progression of prostate cancer. PMID- 12763037 TI - AP-4F, antennapedia peptide linked to an amphipathic alpha helical peptide, increases the efficiency of Lipofectamine-mediated gene transfection in endothelial cells. AB - Typically, endothelial cells are difficult to transfect. In this study, we report that antennapedia peptide (AP) linked to L-4F, a water-soluble, amphipathic alpha helical peptide that avidly binds lipids (AP-4F) increases Lipofectamine 2000 mediated transfection of bovine coronary endothelial cell cultures. Transfection efficiency was monitored by flow cytometry and fluorescent microscopy. Lipofectamine 2000 transfection of endothelial cell cultures with green fluorescence protein (GFP)-DNA typically yields transfection efficiencies of 35.4+/-3.3% with low levels of cell death (8.1+/-1.0%). Pre-treatment of the Lipofectamine 2000-GFP-DNA complexes with AP-4F for 5 min increased transfection to 58.2+/-2.8% without increasing cell death. AP-4F increases Lipofectamine 2000 mediated transfection in a time-dependent fashion (within 10-20 min). Systematic studies reveal that the individual components of AP-4F, i.e., AP and L-4F alone, are ineffective in increasing Lipofectamine 2000-mediated transfection and that AP-4F must be directly associated with DNA liposomes prior to transfection for optimal uptake by endothelial cells. These observations demonstrate that AP-4F may be useful for increasing the transfection efficiency of endothelial cell cultures with standard commercially available reagents. PMID- 12763038 TI - Expression of the activity of cystine/glutamate exchange transporter, system x(c)(-), by xCT and rBAT. AB - The expression of the activity of cystine/glutamate exchange transporter, designated system x(c)(-), requires two components, xCT and 4F2 heavy chain (4F2hc) in Xenopus oocytes. rBAT (related to b(0,+) amino acid transporter) has a significant homology to 4F2hc and is known to be located in the apical membrane of epithelial cells. To determine whether xCT can associate with rBAT and express the activity of system x(c)(-), xCT, and rBAT were co-expressed in Xenopus oocytes and in mammalian cultured cells. In the oocytes injected with rBAT cRNA alone, the activities of cystine and arginine transport were induced, indicating that the system b(0,+)-like transporter was expressed by associating the exogenous rBAT with an endogenous b(0,+)AT-like factor as reported previously. In the oocytes injected with xCT and rBAT cRNAs, the activity of cystine transport was further induced. This induced activity of cystine transport was partially inhibited by glutamate or arginine and completely inhibited by adding both amino acids. In these oocytes, the activity of glutamate transport was also induced and it was strongly inhibited by cystine. In NIH3T3 cells transfected with xCT cDNA alone, the activity of cystine transport was significantly increased, and in the cells transfected with both xCT and rBAT cDNAs, the activity of cystine transport was further enhanced. The enhanced activity was Na(+)-independent and was inhibited by glutamate and homocysteate. These results indicate that rBAT can replace 4F2hc in the expression of the activity of system x(c)(-) and suggest that system x(c)(-) activity could be expressed in the apical membrane of epithelial cells. PMID- 12763039 TI - Sulphite oxidase gene expression in human brain and in other human and rat tissues. AB - Sulphite oxidase (EC 1.8.3.1) is a molybdopterin-containing enzyme that catalyses the oxidation of sulphite to sulphate. Lack of active enzyme produces severe neurodegeneration and early death in humans, showing its essential role. Despite this, the expression of the sulphite oxidase gene in human and rat tissues (especially the brain) has not been elucidated. We therefore examined these tissues and found that the human liver, kidney, skeletal muscle, heart, placenta, and brain showed substantial expression while thymus, spleen, peripheral blood leucocytes, colon, small intestine, and lung showed little expression in humans. In rat, the liver, kidney, heart, brain, and lung (but not skeletal muscle) revealed a hybridization signal with the strongest signal in the liver. The spleen and testis also showed little expression. The differential expression of sulphite oxidase gene in various human brain regions was studied. Expression was seen in all brain regions examined (cerebellum, cerebral cortex, medulla, spinal cord, occipital pole, frontal lobe, amygdala, caudate nucleus, corpus callosum, hippocampus, thalamus, temporal lobe, and putamen). The cerebral cortex showed the highest level of expression. PMID- 12763040 TI - In vitro-generated respiratory mucosa: a new tool to study inhalational anthrax. AB - We generated a three-dimensional (3-D) model of human airway tissues in order to study initiation of inhalational form of anthrax infection. The system was designed to model the air-blood barrier of the respiratory tract represented by epithelial cells and macrophages. When grown on collagen/fibronectin gel support at an air-liquid interface, airway epithelial cells formed cell layers morphologically resembling those in vivo. These preformed epithelial cell cultures were further supplemented with monocytes/macrophages isolated from human blood. After 2-5 days of co-culture, monocytes differentiated into a phenotype of resident macrophages, which was evaluated by the expression of specific cell surface markers. This model allowed sorting out the role of each type of cell found at the air surface of the lung. The interdependence of macrophages and epithelial cells in the clearance of anthrax spores from airways and the capacity of the airway epithelial cells to protect from anthrax infection was demonstrated. PMID- 12763041 TI - Activation of in situ tissue transglutaminase by intracellular reactive oxygen species. AB - We have investigated the novel function of intracellular reactive oxygen species (ROS) in the activation of in situ tissue transglutaminase (tTGase) by lysophosphatidic acid (LPA) and transforming growth factor-beta (TGF-beta) in Swiss 3T3 fibroblasts. LPA induced a transient increase of intracellular ROS with a maximal increase at 10 min, which was blocked by ROS scavengers, N-acetyl-L cysteine and catalase. LPA activated tTGase with a maximal increase at 1h, which was inhibited by cystamine and ROS scavengers. Incubation with exogenous H(2)O(2) activated tTGase. TGF-beta also activated tTGase with a maximal activation at 2h and the tTGase activation was inhibited by the ROS scavengers. Scrape-loading of C3 transferase inhibited the ROS production and in situ tTGase activation by LPA and TGF-beta, and the inhibitory effect of C3 transferase was reversed by exogenous H(2)O(2). Microinjection of GTPgammaS inhibited transamidating activity of tTGase stimulated by LPA, TGF-beta, and maitotoxin. These results suggested that intracellular ROS was essential for the activation of in situ tTGase in response to LPA and TGF-beta. PMID- 12763042 TI - Only one dnaK homolog, dnaK2, is active transcriptionally and is essential in Synechocystis. AB - We examined the expression and the function of the DnaK chaperone family in the photoautotrophic cyanobacterium, Synechocystis PCC 6803. Surprisingly, only one of the three dnaK genes was transcribed either under normal or heat shock conditions. Their predicted cochaperones (four dnaJs and one grpE) proved to be uninducible under our experimental conditions. Attempts to inactivate the active dnaK2 has failed, indicating that the gene is essential. The partial mutant displayed lower inducibility of chaperones (especially GroEL and HSP17) both at mRNA and protein levels upon heat shock. The mutant showed temperature sensitive phenotype, but was able to acquire thermotolerance. PMID- 12763043 TI - Gene expression of Toll-like receptors and associated molecules induced by inflammatory stimuli in the primary alveolar macrophage. AB - We investigated the expression of Toll-like receptors (TLRs) and associated signaling molecules in inflammatory stimuli-activated murine primary alveolar macrophage (AM) in vitro, and in a murine model of acute lung injury. The results demonstrated three patterns of gene expression: the TLR2 and myeloid differentiation factor 88 (MyD88) gene expressions were induced in AM in response to lipopolysaccharide (LPS), interleukin (IL)-1beta, or tumor necrosis factor alpha or in the lung tissue of an LPS-induced acute lung injury model; the gene expressions of TLR1, -3, -6, CD14, and MD2 were unchanged; and the TLR4 and TLR5 gene expressions were downregulated in AM following inflammatory stimuli. Furthermore, the LPS-induced upregulation of the TLR2 gene was mediated via the activation of NF-kappaB. These results indicate that the TLR2 expression upregulated in AM following bacterial respiratory infections may render AM responsive to TLR2 ligands, which may enhance innate immunity against pathogens in the lung. PMID- 12763044 TI - Ascorbic acid spares alpha-tocopherol and decreases lipid peroxidation in neuronal cells. AB - Ascorbic acid is considered an antioxidant in the central nervous system, but direct evidence that ascorbate protects neuronal cells from oxidant stress is lacking. Differentiated SH-SY5Y cells in culture took up ascorbic acid on the sodium-dependent vitamin C transporter Type 2 and retained it much more effectively than dehydroascorbic acid. Intracellular ascorbate spared alpha tocopherol, both in cells loaded with alpha-tocopherol in culture and in cells under oxidant stress due to extracellular ferricyanide. Sparing of alpha tocopherol in response to ferricyanide was associated with protection against lipid peroxidation in cell membranes. These results show that neuronal cells concentrate ascorbate, and that intracellular ascorbate, either directly or through sparing of alpha-tocopherol, protects them against oxidant stress. PMID- 12763045 TI - Induction of phase 2 enzymes by serum oxidized polyamines through activation of Nrf2: effect of the polyamine metabolite acrolein. AB - The naturally occurring polycationic polyamines including putrescine, spermidine, and spermine play an important role in cell growth, differentiation, and gene expression. However, circulating polyamines are potential substrates for several oxidizing enzymes including copper-containing serum amine oxidase. These enzymes are capable of oxidizing serum polyamines to several toxic metabolites including aldehydes and H(2)O(2). In this study, we investigated the effects of polyamines as inducers of phase 2 enzymes and other genes that promote cell survival in a cell culture system in the presence of bovine serum. Spermidine and spermine (50 microM) increased NAD(P)H quinone oxidoreductase (NQO1) activity up to 3-fold in murine keratinocyte PE cells. Transcript levels for glutathione S-transferase (GST) A1, GST M1, NQO1, gamma-glutamylcysteine ligase regulatory subunit, and UDP glucuronyltransferase 1A6 were significantly increased by spermidine and this effect was mediated through the antioxidant response element (ARE). The ARE from the mouse GST A1 promoter was activated about 9-fold by spermine and 5-fold by spermidine treatment, but could be inhibited by the amine oxidase inhibitor, aminoguanidine, suggesting that acrolein or hydrogen peroxide generated from polyamines by serum amine oxidase may be mediators for phase 2 enzyme induction. Elevations of ARE-luciferase expression and NQO1 enzyme activity by spermidine were not affected by catalase, while both were completely repressed by aldehyde dehydrogenase treatment. Direct addition of acrolein to PE cells induced multiple phase 2 genes and elevated nuclear levels of Nrf2, a transcription factor that binds to the ARE. Expression of mutant Nrf2 repressed the activation of the ARE luciferase reporter by polyamines and acrolein. These results indicate that spermidine and spermine increase the expression of phase 2 genes in cells grown in culture through activation of the Nrf2-ARE pathway by generating the sulfhydryl reactive aldehyde, acrolein. PMID- 12763046 TI - O2 can raise fetal pneumocyte Na+ conductance without affecting ENaC mRNA abundance. AB - In fetal pneumocytes, increasing P(O(2)) can raise apical Na(+) conductance (G(Na(+))) and increase the abundance of epithelial Na(+) channel subunit (alpha , beta-, and gamma-ENaC) mRNA, suggesting that the rise in G(Na(+)), which may be important to the perinatal maturation of the lung, reflects O(2)-evoked ENaC gene expression. However, we now show that physiologically relevant increases in P(O(2)) do not affect alpha-, beta-, and gamma-ENaC mRNA abundance in pneumocytes maintained (approximately 48 h) in hormone-free medium or in medium supplemented with dexamethasone and tri-iodothyronine, although the response does persist in cells maintained in medium containing a complex mixture of hormones/growth factors. However, parallel electrometric studies revealed clear increases in G(Na(+)) under all tested conditions and so it is now clear that O(2)-evoked increases in G(Na(+)) can occur without corresponding increases in ENaC mRNA abundance. It is therefore unlikely that this rise in G(Na(+)) is secondary to O(2)-evoked ENaC gene expression. PMID- 12763047 TI - Mechanotransduction in bone cells proceeds via activation of COX-2, but not COX 1. AB - Cyclooxygenase (COX) is the key enzyme in the production of prostaglandins, which are essential for the response of bone to mechanical loading. We determined which COX-isoform, COX-1 or COX-2, determines loading-induced prostaglandin production in primary bone cells in vitro. Mouse and human bone cells reacted to 1 h of pulsating fluid flow (PFF, 0.6+/-0.3 Pa at 5 Hz) with an increased prostaglandin E(2) production, which continued 24 h after cessation of PFF. Inhibition of COX-2 activity with NS-398 abolished the stimulating effect of PFF both at 1 h and at 24 h post-incubation, while inhibition of COX-1 by SC-560 affected neither the early nor the late response to flow. PFF rapidly stimulated COX-2 mRNA expression at 1 h but did not affect COX-1 mRNA expression. COX-2 mRNA expression was still significantly enhanced 24 h after cessation of PFF. We conclude that COX-2 is the mechanosensitive form of COX that determines the response of bone tissue to mechanical loading. PMID- 12763048 TI - Smad3 is required for enamel biomineralization. AB - Smad3 is an intracellular signaling molecule that mediates the signal from transforming growth factor-beta (TGF-beta) and activin receptors. In this study, we reveal hypomineralized enamel in mice with the targeted deletion of the Smad3 gene. The Smad3 (-/-) mice had chalky white incisor enamel, while the enamel of the wild-type or Smad3 (+/-) mice was yellow-brown. Histological analysis of the undecalcified sections showed that the enamel thickness of the maxillary incisors in the Smad3 (-/-) mice was similar to that of the wild-type and Smad3 (+/-) mice while that the enamel of the maxillary molars in Smad3 (-/-) mice was disrupted in places. Microcomputed tomography (microCT) analysis revealed that the mineralization of the maxillary incisors and mandibular molars in the Smad3 (-/-) mice showed significant reduction in the degree of mineralization when compared to that of the wild-type and Smad3 (+/-) mice. Scanning electron microscopic (SEM) analysis of the mandibular incisors revealed that the enamel surface of the Smad3 (-/-) mice was irregular and disrupted in places and showed images similar to decalcified mature enamel. The histological analysis of the decalcified sections showed that distinct morphological changes in the ameloblasts at the secretory and maturational stages were not observed between the Smad3 (-/-) and Smad3 (+/-) or wild-type mice, while the enamel matrix was observed in the decalcified sections of the mandibular molars in the Smad3 (-/-) mice. These results suggested that Smad3 was required for enamel biomineralization, and TGF beta and activin signaling might be critical for its process. PMID- 12763049 TI - The maintenance of neuromuscular function requires UBC-25 in Caenorhabditis elegans. AB - Caenorhabditis elegans gene ubc-25 encodes a novel type of an E2 ubiquitin transferase domain (UBCc) protein, which is highly conserved in multicellular animals, but which is not present in the genomes of fungi or plants. To identify the cellular localization of UBC-25 during the development of C. elegans, we used a ubc-25::gfp reporter gene construct. These experiments showed that ubc-25 expression starts during embryogenesis and that it is restricted to neurons and muscle cells in all later stages of development as well as in adult animals. RNA interference with ubc-25 caused late-onset paralysis of most muscular functions such as locomotion, egg laying, and defecation. We therefore propose that ubc-25 in C. elegans is required for the maintenance (homeostasis) of neuromuscular functions by contributing to a tissue specific protein modification pathway, and we speculate that the adult onset phenotype results from the accumulation of target proteins which fail to be degraded. PMID- 12763050 TI - Increased gene expression and production of murine endothelin receptors after birth. AB - We developed the real-time PCR quantification of endothelin-A (ET-A) and endothelin-B (ET-B) receptor genes and present their relative expression levels in various adult tissues and during development in mouse using the 2(-Delta Delta C(T)) method. ET-A and ET-B receptors were detected in all tissues examined. Gene expression of ET-A and ET-B receptors increases during the later stages of embryonic development in lung, heart, liver, kidney, and skin and reaches a maximum on the first one or two days after birth. The results, in agreement with our data on endothelin (ET) ligands, suggest that the ET system may be involved in the emergence and maintenance of functions vital after birth in these organs. These findings were corroborated through observation of the correlation between the gene expression and (poly)peptide production of the ET system in normal skin before and after parturition. PMID- 12763051 TI - Selective degradation of oxidatively modified protein substrates by the proteasome. AB - Oxidative stress in mammalian cells is an inevitable consequence of their aerobic metabolism. Oxidants produce modifications to proteins leading to loss of function (or gain of undesirable function) and very often to an enhanced degradation of the oxidized proteins. For several years it has been known that the proteasome is involved in the degradation of oxidized proteins. This review summarizes our knowledge about the recognition of oxidized protein substrates by the proteasome in in vitro systems and its applicability to living cells. The majority of studies in the field agree that the degradation of mildly oxidized proteins is an important function of the proteasomal system. The major recognition motif of the substrates seems to be hydrophobic surface patches that are recognized by the 20S 'core' proteasome. Such hydrophobic surface patches are formed by partial unfolding and exposure of hydrophobic amino acid residues during oxidation. Oxidized proteins appear to be relatively poor substrates for ubiquitination, and the ubiquitination system does not seem to be involved in the recognition or targeting of oxidized proteins. Heavily oxidized proteins appear to first aggregate (new hydrophobic and ionic bonds) and then to form covalent cross-links that make them highly resistant to proteolysis. The inability to degrade extensively oxidized proteins may contribute to the accumulation of protein aggregates during diseases and the aging process. PMID- 12763052 TI - Ozone-induced disruptions of lung transcriptomes. AB - We have analyzed changes in approximately 4000 lung mRNAs, with GeneChips, in mice exposed to 1 ppm O(3) for three consecutive nights (8 h per night). Differential gene expression analysis identified approximately 260 O(3) sensitive genes; approximately 80% of these were repressed and approximately 20% were induced in O(3)-exposed mice compared to the air-exposed controls. A 20-fold induction of serum amyloid A3 mRNA by O(3) suggested activation of NF-kappaB and CCAAT/enhancer binding protein-mediated pathways by inflammatory cytokines. Induction (up to 14-fold) of 12 genes that increase DNA synthesis and cell cycle progression, and increase (approximately 7-fold) in CD44 mRNA and macrophage metalloelastase suggested a state of O(3)-induced hyperplasia and lung remodeling. Several mRNAs encoding enzymes of xenobiotic metabolism and cytoskeletal functions were repressed and may suggest cytokine mediated suppression of cytochrome P450 expression and cachexia-like inflammatory state in ozone-exposed lungs. The expressions of approximately 30 genes of immune response were also repressed. Collectively this genome-wide analysis of lungs identified ozone-induced disruption of gene transcriptional profile indicative of increased cellular proliferation under suppressed immune surveillance and xenobiotic metabolism. PMID- 12763053 TI - Radical-radical reactions of superoxide: a potential route to toxicity. AB - Superoxide reacts with many radicals, such as phenoxyl radicals, at near diffusion-controlled rates. These reactions are usually considered to be repair processes and have received little biological attention. However, addition of superoxide to give hydroperoxides and secondary oxidation products can also occur. The relative contributions of addition and repair vary depending on the properties of the phenol. With tyrosine, addition to give tyrosine hydroperoxide predominates, but in peptides the efficiency of hydroperoxide formation depends on the proximity of free amine groups. Radicals from other phenolic compounds, such as alpha-tocopherol and serotonin, also undergo addition reactions with superoxide. Physiologically, these reactions are likely to be more significant than dimerization when both radicals are generated together. They warrant attention as potential contributors to superoxide toxicity. PMID- 12763054 TI - Vitamin E and drug metabolism. AB - Tocopherols and tocotrienols are metabolized by side chain degradation initiated by cytochrome P450 (CYP)-catalyzed omega-hydroxylation followed by beta oxidation. Whereas alpha-tocopherol is only poorly metabolized, high amounts of the final products, carboxyethyl hydroxychroman (CEHC), are found from other tocols in HepG2 cells and in human urine. CYP3A4 and CYP4F2 were suggested to be involved in tocopherol degradation. CYP3A4 metabolizes most of the drugs and is induced by many of its substrates via the activation of the pregnane X receptor (PXR). Also tocopherols and in particular tocotrienols induce the expression of a PXR-driven reporter gene and the expression of endogenous CYP3A4 and CYP3A5 which is supported by sporadic publications spread over the last 30 years. The potential interference of vitamin E with drug metabolism is discussed in the light of related complications evoked by herbal remedies. PMID- 12763055 TI - Induction of stress proteins and MMP-9 by 0.8 ppm of ozone in murine skin. AB - Ozone (O(3)) is among the most reactive environmental oxidant pollutants to which cutaneous tissues are exposed. O(3) exposure has been shown to induce antioxidant depletion as well as the oxidation of lipids and proteins within the outermost skin layer, the stratum corneum. However, relatively little is known regarding the potential effects of O(3) on the cellular constituents of the underlying skin epidermis and dermis. In the present study, hairless mice exposed for 6 h to 0.8 ppm O(3) showed increases in lipid peroxidation, as quantitated by increases in 4 hydroxynonenal-protein adducts. O(3) exposure caused an induction of the stress proteins HSP27 and heme oxygenase-1 (HO-1), starting at 6 h and increasing up to 18 h after O(3) exposure. This was accompanied by an increase in matrix metalloproteinase-9 (MMP-9) mRNA and activity levels, indicative of possible injurious-reparative processes. Collectively, our data demonstrate that skin exposure to O(3) not only affects antioxidant levels and oxidation markers in the outermost stratum corneum layer, but also induces cellular stress responses in the deeper cellular layers of the skin. PMID- 12763056 TI - Susceptibility of plasma lipids to peroxidation. AB - Lipid peroxidation is an old and yet novel subject. It induces membrane disturbance and damage and its products are known to induce the generation of various cytokines and cell signaling. In the present work, the susceptibility and specificity of human plasma lipids to oxidation were studied, aiming specifically at elucidating the effects of oxidation milieu and oxidants. Cholesteryl esters (CEs) and phosphatidylcholines (PCs) were more readily oxidized in plasma than in organic solution under similar conditions. The susceptibilities of PC and free cholesterol (FC) relative to CE to free radical-mediated lipid peroxidation induced by peroxyl radicals and peroxynitrite were smaller in plasma than in organic solution. The higher rate of CE oxidation by free radicals than PC may be accounted for by the physical effects as well as higher content of polyunsaturated lipids in CE than PC. On the contrary, PC was more readily oxidized than CE by lipoxygenases. The lipid hydroperoxides were stable in organic solution but reduced to the corresponding hydroxides in plasma, the rate being much faster for PC hydroperoxides than for CE and FC hydroperoxides. It was confirmed that free radical-mediated oxidation gave both cis,trans and trans,trans, racemic, random hydroperoxides, while that by lipoxygenase gave only regio- and stereo-specific cis,trans-hydroperoxide. PMID- 12763057 TI - Carotenoid-radical interactions. AB - Carotenoids have been reported to react with virtually any radical species likely to be encountered in a biological system. The products of such reactions are frequently short-lived radical species that can decay to more stable products. In some cases, stable adducts can be observed, but in the majority of interactions with radicals, carotenoids break down to degradation products very similar to what is seen with oxidative degradation. It is only recently that the biological activity of these breakdown products has begun to be investigated. PMID- 12763058 TI - Singlet oxygen-mediated damage to proteins and its consequences. AB - Proteins comprise approximately 68% of the dry weight of cells and tissues and are therefore potentially major targets for oxidative damage. Two major types of processes can occur during the exposure of proteins to UV or visible light. The first of these involves direct photo-oxidation arising from the absorption of UV radiation by the protein, or bound chromophore groups, thereby generating excited states (singlet or triplets) or radicals via photo-ionisation. The second major process involves indirect oxidation of the protein via the formation and subsequent reactions of singlet oxygen generated by the transfer of energy to ground state (triplet) molecular oxygen by either protein-bound, or other, chromophores. Singlet oxygen can also be generated by a range of other enzymatic and non-enzymatic reactions including processes mediated by heme proteins, lipoxygenases, and activated leukocytes, as well as radical termination reactions. This paper reviews the data available on singlet oxygen-mediated protein oxidation and concentrates primarily on the mechanisms by which this excited state species brings about changes to both the side-chains and backbone of amino acids, peptides, and proteins. Recent work on the identification of reactive peroxide intermediates formed on Tyr, His, and Trp residues is discussed. These peroxides may be important propagating species in protein oxidation as they can initiate further oxidation via both radical and non-radical reactions. Such processes can result in the transmittal of damage to other biological targets, and may play a significant role in bystander damage, or dark reactions, in systems where proteins are subjected to oxidation. PMID- 12763059 TI - Oxygen dependence of mitochondrial nitric oxide synthase activity. AB - The effect of O(2) concentration on mitochondrial nitric oxide synthase (mtNOS) activity and on O(2)(-) production was determined in rat liver, brain, and kidney submitochondrial membranes. The K(mO(2)) for mtNOS were 40, 73, and 37 microM O(2) and the V(max) were 0.51, 0.49, and 0.42 nmol NO/minmg protein for liver, brain, and kidney mitochondria, respectively. The rates of O(2)(-) production, 0.5-12.8 nmol O(2)(-)/minmg protein, depended on O(2) concentration up to 1.1mM O(2). Intramitochondrial NO, O(2)(-), and ONOO(-) steady-state concentrations were calculated for the physiological level of 20 microM O(2); they were 20-39 nM NO, 0.17-0.33 pM O(2)(-), and 0.6-2.2 nM ONOO(-) for the three organs. These levels establish O(2)/NO ratios of 513-1000 that correspond to physiological inhibitions of cytochrome oxidase by intramitochondrial NO of 16-25%. The production of NO by mtNOS appears as a regulatory process that modulates mitochondrial oxygen uptake and cellular energy production. PMID- 12763060 TI - Biological selectivity and functional aspects of protein tyrosine nitration. AB - The formation of nitric oxide in biological systems has led to the discovery of a number of post-translational protein modifications that could regulate protein function or potentially be utilized as transducers of nitric oxide signaling. Principal among the nitric oxide-mediated protein modifications are: the nitric oxide-iron heme binding, the S-nitrosylation of reduced cysteine residues, and the C-nitration of tyrosine and tryptophan residues. With the exception of the nitric oxide binding to heme iron proteins, the other two modifications appear to require secondary reactions of nitric oxide and the formation of nitrogen oxides. The rapid development of analytical and immunological methodologies has allowed for the quantification of S-nitrosylated and C-nitrated proteins in vivo revealing an apparent selectivity and specificity of the proteins modified. This review is primarily focused upon the nitration of tyrosine residues discussing parameters that may govern the in vivo selectivity of protein nitration, and the potential biological significance and clinical relevance of this nitric oxide mediated protein modification. PMID- 12763062 TI - Endolymphatic sodium homeostasis by Reissner's membrane. AB - Cochlear sensory transduction depends on active extrusion of sodium ion (Na(+)) from the luminal fluid, endolymph. Reissner's membrane epithelium forms much of the barrier between cochlear endolymph and perilymph and we hypothesized that Reissner's membrane might be responsible for this function. We found that Reissner's membrane isolated from gerbil produced a short circuit current (I(sc)) directed into the apical side, consistent with cation absorption and/or anion secretion. I(sc) was inhibited by amiloride analogs in the potency sequence benzamil>amiloride>>ethylisopropylamiloride, consistent with Na(+) absorption through an epithelial sodium channel in the apical cell membrane. I(sc) was also inhibited by an inhibitor of Na(+),K(+)-ATPase, ouabain, and by the K(+) channel blockers Ba(2+), 4-aminopyridine and quinine but not tetraethylammonium nor glibenclamide, consistent with the presence of a voltage-activated K(+) channel. Bumetanide, an inhibitor of the Na(+),2Cl(-),K(+)-cotransporter, had no effect on I(sc). Contrary to previous hypotheses, no evidence was found for electrogenic secretion of Cl(-) under control of cAMP since neither forskolin nor genistein affected I(sc) when Na(+) absorption was blocked. These results provide the first direct evidence that Reissner's membrane contributes to normal cochlear function by absorption of Na(+) from endolymph. PMID- 12763063 TI - Involvement of cholinergic and gabaergic systems in the fragile X knockout mice. AB - Fragile X syndrome is an inherited cause of mental retardation. We used extra- and intracellular recordings in brain slices obtained from wild type and fragile X knockout mice to establish whether bath application of the cholinergic agent carbachol (5 microM) induces different responses in neurons of the subiculum, a limbic structure involved in learning and memory. We found that carbachol diminished excitatory post-synaptic responses induced by CA1 stratum radiatum stimulation in wild type mice, but caused an unexpected increase in knockout animals. Moreover, these responses augmented in knockout mice after carbachol washout, a phenomenon that resembled the muscarinic long-term potentiation seen in wild type mice during application of carbachol and GABA(A) receptor antagonists. We also used paired-pulse stimulation to determine whether the changes in synaptic excitability induced by carbachol were caused by pre- or post synaptic mechanism. Under control conditions, this protocol induced facilitation in both wild type and knockout mice; in contrast, during carbachol application, this facilitatory effect was seen in wild type mice only. In conclusion, our data highlight for the first time differences in cholinergic and GABA-ergic mechanisms that may contribute to the phenotype of fragile X patients. PMID- 12763064 TI - Selective blockade of gene expression in a single identified snail neuron. AB - In the present study, the applicability of antisense morpholino oligos for loss of-function experiments in neurobiology was investigated. The identified withdrawal interneurons of the parietal ganglia expressing helix command neuron specific 2 (HCS2) gene were pressure injected with HCS2 antisense or control morpholino oligo solution at a final concentration 1-4 microM. No toxic or side effects for the neural functioning were noted immediately or several hours after injection. The changes in the concentration of HCS2-encoded protein in neurons after injection were monitored by two methods, Western blotting and immunostaining of the brain. The amount of the peptide immunoreactive with the HCS2 antibody started to decline in the injected cells at day 2 post-injection, decreased four- to five-fold at day 4, and stayed at this low level thereafter. Similar results obtained by both methods suggest significant selective blockade of production of the HCS2-encoded peptide. In contrast, no substantial decrease of the HCS2-encoded polypeptide was observed after injection with control oligos. Due to the high stability of the morpholino oligos in the cell, they represent a highly efficient tool for a specific long-term blockade of gene expression in molluscan neurons. PMID- 12763065 TI - Individual nucleus accumbens-projection neurons receive both basolateral amygdala and ventral subicular afferents in rats. AB - The nucleus accumbens is regarded as the limbic-motor interface, in view of its limbic afferent and somatomotor and autonomic efferent connections. Within the accumbens, there appear to be specific areas in which limbic afferent fibres, derived from the hippocampus and the amygdala, overlap. These afferent inputs have been suggested to converge monosynaptically on cells within the accumbens and are hypothesized to play a role in paradigms such as conditioned place preference. Convergence between inputs from basolateral amygdala and hippocampus can be demonstrated with electrophysiological recording methods, but these do not conclusively preclude polysynaptic mechanisms. We examined the synaptic input to the projection neurons of the accumbens, the medium-sized densely spiny neurons. We labelled the projection neurons with a small injection of biotinylated dextran amine into the accumbens, and the afferents from the basolateral amygdala and ventral subiculum of the hippocampus with injections of biotinylated dextran amine and Phaseolus vulgaris-leucoagglutinin respectively, and revealed the anterogradely labelled fibres with different chromogens. The labelled accumbens projection neurons were studied with correlated light and electron microscopy for identified monosynaptic inputs. With this technique we have demonstrated anatomically that monosynaptic convergence between the ventral subicular region of the hippocampus and the basolateral region of the amygdala occurs at the level of the proximal as well as distal dendrites. Finally, we suggest that these anatomical arrangements may represent the framework for the integrative role that has been assigned to the accumbens. PMID- 12763066 TI - Ligand-induced mu opioid receptor endocytosis and recycling in enteric neurons. AB - Immunohistochemistry and confocal microscopy were used to investigate endocytosis and recycling of the native mu opioid receptor (muOR) in enteric neurons. Isolated segments of the guinea-pig ileum were exposed to increasing concentrations of muOR agonists at 4 degrees C to allow ligand binding and warming to 37 degrees C for 0 min (baseline) to 6 h in ligand-free medium to allow receptor internalization and recycling. The endogenous ligand, [Met]enkephalin, and [D-Ala(2),MePhe(4),Gly-ol(5)] enkephalin (DAMGO), an opioid analog, and the alkaloids, etorphine and fentanyl, induced rapid internalization of muOR immunoreactivity in enteric neurons, whereas morphine did not. muOR internalization was prevented by muOR antagonists. Basal levels of muOR immunoreactivity in the cytoplasm were 10.52+/-2.05%. DAMGO (1 nM-100 microM) induced a concentration-dependent increase of muOR immunofluorescence density in the cytoplasm to a maximum of 84.37+/-2.26%. Translocation of muOR immunoreactivity in the cytoplasm was detected at 2 min, reached the maximum at 15-30 min, remained at similar levels for 2 h, began decreasing at 4 h, and was at baseline values at 6 h. A second exposure to DAMGO (100 nM) following recovery of internalized muOR immunoreactivity at the cell surface induced a translocation of muOR immunoreactivity in the cytoplasm comparable to the one observed following the first exposure (46.89+/-3.11% versus 43.31+/-3.80%). muOR internalization was prevented by hyperosmolar sucrose, phenylarsine oxide or potassium depletion, which inhibit clathrin-mediated endocytosis. muOR recycling was prevented by pre-treatment with bafilomycin A1, an acidotropic agent that inhibits endosomal acidification, but not by the protein synthesis inhibitor, cycloheximide. This study shows that native muOR in enteric neurons undergoes ligand-selective endocytosis, which is primarily clathrin-mediated, and recycles following endosomal acidification. Following recycling, muOR is activated and internalized by DAMGO indicating that recycled receptors are functional. PMID- 12763067 TI - Connective tissue growth factor: a novel marker of layer VII neurons in the rat cerebral cortex. AB - Connective tissue growth factor (CTGF) belongs to a family of secreted, extracellular matrix-associated proteins that are involved in the regulation of cellular functions such as adhesion, migration, mitogenesis, differentiation and survival. Recent studies have also suggested the up-regulation of CTGF in response to trauma, scar formation and excitotoxicity in the CNS. To further elucidate the localization and regulation of this molecule in the rat brain we performed in situ hybridization experiments and found a very strong and selective expression of CTGF messenger ribonucleic acid (mRNA) on the band of layer VII neurons throughout the adult cerebral cortex. Similarly strong neuronal expression was also present in the dorsal endopiriform nucleus, extending rostrally from the ventrocaudal cortical layer VII, and in the deep layers of the olfactory glomeruli and the accessory olfactory nucleus. Double in situ hybridization confirmed selective CTGF mRNA expression on a subpopulation (approximately 35%) of microtubule-associated protein 2 mRNA-positive neurons in the cortical layer VII and the dorsal endopiriform nucleus. The nucleus of lateral olfactory tract showed moderate signal intensity; other parts of the forebrain, mesencephalon and brain stem only revealed a very weak level of CTGF mRNA expression. Non-neuronal expression was rare, considerably weaker than on cortical layer VII neurons, and normally associated with blood vessels. Developmental analysis of CTGF mRNA expression in embryonic and postnatal mouse also showed a moderately late onset at embryonic day 16-18, and confirmed the presence of CTGF mRNA in cortical layer VII in a second rodent species. Interestingly, injury experiments using direct cerebral trauma or injection of excitotoxic kainic acid into rat brain failed to up-regulate CTGF mRNA after injury and during the ensuing period of neuronal cell death, gliosis and neural scar tissue formation. Altogether, the current data suggest a constitutive role of CTGF, particularly in the adult cerebral cortex. In view of the strong ascending projections of subplate neurons into cortical layer 1, this molecule may be involved in the modulation of synaptic input to apical dendrites of pyramidal neurons. PMID- 12763069 TI - Prostaglandin E2 inhibits the potassium current in sensory neurons from hyperalgesic Kv1.1 knockout mice. AB - Prostaglandin E(2) (PGE(2)) enhances the sensitivity of sensory neurons to various forms of noxious stimulation. This occurs, in part, by the suppression of a delayed rectifier-like potassium current in these neurons. However, the molecular identity of this current remains unclear. Recent studies demonstrated that a mutant mouse lacking a delayed rectifier potassium channel gene, Kv1.1, displayed lowered thresholds to thermal stimulation in behavioral assays of pain perception, i.e. the Kcna1-null mice were hyperalgesic. Here we examined whether PGE(2) can alter the sensitivity of Kcna1-null mice to noxious stimulation and examine the capability of PGE(2) to inhibit the potassium current in these knockout mice. Behavioral assays were used to assess the effect of PGE(2) on either thermal hyperalgesia or mechanical sensitivities. In addition, the whole cell patch-clamp technique was used to study the effects of PGE(2) on the total potassium current recorded from isolated mouse sensory neurons. Even with a reduced threshold to thermal stimulation, PGE(2) could still sensitize the response of Kcna1-null mice to thermal and mechanical stimulation by amounts that were similar to that in wild type mice. The activation properties of the potassium current were similar for both the wild type and the Kcna1-null mice, whereas the inactivation properties were different in cells exhibiting large amounts of steady-state inactivation (>50%) measured at +20 mV. PGE(2) suppressed the total potassium current in both groups of mice by 40-50% without altering the voltage dependence of activation. In addition, PGE(2) produced similar amounts of suppression in both groups of mice when currents were examined with the steady state inactivation protocol. Based on these results, it is unlikely that Kv1.1 is the molecular identity of the potassium channel(s) modulated by PGE(2) to sensitize nociceptive sensory neurons. Also, the enhanced thermal sensitivity as observed in the Kcna1-null mice might be due to more central neurons of the pain sensing pathway. PMID- 12763068 TI - Serotonin type II receptor activation facilitates synaptic plasticity via N methyl-D-aspartate-mediated mechanism in the rat basolateral amygdala. AB - The modulation of synaptic plasticity by serotonin type II (5-hydroxytryptamine type II (5-HT(2)))-receptor stimulation was explored using intracellular, field potential and Fura-2 fluorescence image recordings in a rat amygdala slice preparation. Bath application of 5HT(2) receptor agonist 1-(2,5)-dimethoxy-4 iodophen-2-aminopropane (DOI) transformed theta-burst-stimulated (TBS) synaptic plasticity from short-term potentiation to long-term potentiation. DOI enhanced N methyl-D-aspartate (NMDA) receptor-mediated potentials and calcium influx without affecting the resting membrane potential or input resistance of the neurons. In contrast, alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA)/kainate receptor-mediated excitatory synaptic responses were unaffected by DOI. The facilitating effects of DOI were blocked by the 5-HT(2) receptor antagonist, ketanserin, and by the 5-HT(2C)-receptor selective antagonist, RS102221. These results indicate that 5-HT(2)-receptor activation enhances NMDA receptor-mediated synaptic function in the basolateral amygdala (BLA). PMID- 12763070 TI - Sec6 is localized to the plasma membrane of mature synaptic terminals and is transported with secretogranin II-containing vesicles. AB - The sec6/8 (exocyst) complex is implicated in targeting of vesicles for regulated exocytosis in various cell types and is believed to play a role in synaptogenesis and brain development. We show that the subunits sec6 and sec8 are present at significant levels in neurons of adult rat brain, and that immunoreactivity for the two subunits has a differential subcellular distribution. We show that in developing as well as mature neurons sec6 is concentrated at the inside of the presynaptic plasma membrane, while sec8 immunoreactivity shows a diffuse cytoplasmic distribution. Among established, strongly synaptophysin-positive neuronal boutons, sec6 displays highly differential concentrations, indicating a role for the complex independent of the ongoing synaptic-vesicle release activity. Sec6 is transported along neurites on secretogranin II-positive vesicles, while sec6-negative/secretogranin II-positive vesicles stay in the cell body. In PC12 cells, sec6-positive vesicles accumulate at the plasma membrane at sites of cell-cell contact. Neuronal induction of the PC12 cells with nerve growth factor shows that sec8 is not freely soluble, but may probably interact with cytoskeletal elements. The complex may facilitate the targeting of membrane material to presynaptic sites and may possibly shuttle vesicles from the cytoskeletal transport machinery to presynaptic membrane sites. Thus, we suggest that the exocyst complex serves to modulate exocytotic activity, by targeting membrane material to its presynaptic destination. PMID- 12763071 TI - Coupling of neuronal activity and mitochondrial metabolism as revealed by NAD(P)H fluorescence signals in organotypic hippocampal slice cultures of the rat. AB - During physiological activity neurons may experience localised energy demands which require intracellular signals for stimulation of mitochondrial NADH generation and subsequent delivery of ATP. To elucidate these mechanisms, we applied microfluorimetric monitoring of cytoplasmic (Fluo-3) and mitochondrial (Rhod-2) calcium concentration ([Ca(2+)](c), [Ca(2+)](m)), as well as of mitochondrial oxidative metabolism (NAD(P)H), whilst simultaneously measuring changes in extracellular potassium concentration ([K(+)](o)), as an indicator of neuronal activity in hippocampal slice cultures. Changes in neuronal activity were induced by repetitive stimulation at different frequencies (5, 20, 100 Hz) and intensities. Stimulation parameters were chosen to elicit rises in [K(+)](o) of less than 3 mM which is comparable to physiologically occurring rises in [K(+)](o). The mitochondrial uncoupler carbonyl cyanide m-chlorophenyl hydrazone (CCCP) reduced stimulus-induced changes in Rhod-2 fluorescence by 79%, indicating that Rhod-2 signals were primarily of mitochondrial origin. Repetitive stimulation at 20 Hz applied to areas CA1 or CA3 resulted in moderate rises in [K(+)](o) which were associated with stimulus-dependent elevations in [Ca(2+)](c) and [Ca(2+)](m) of up to 15%. The same stimuli also elicited biphasic changes in NAD(P)H fluorescence characterised by an initial decline and a subsequent prolonged elevation of up to 10%. Variation of stimulus parameters revealed close correlations between rises in [K(+)](o), in [Ca(2+)](m) and changes in NAD(P)H fluorescence. To elucidate the role of intracellular Ca(2+) accumulation in induction of NAD(P)H fluorescence signals, the effect of application of Ca(2+) free solution on these signals evoked by repetitive antidromic stimulation of the alveus during recordings in area CA1 was studied. Lowering extracellular Ca(2+) led to complete blockade of postsynaptic field potential components as well as of rises in [Ca(2+)](c) and [Ca(2+)](m). Amplitudes of NAD(P)H signals were reduced by 59%, though rises in [K(+)](o) were comparable in presence and absence of extracellular Ca(2+). The results suggest i) that mitochondrial oxidative metabolism is fine-tuned to graded physiological activity in neurons and ii) that rapid mitochondrial Ca(2+) signalling represents one of the main determinants for stimulation of oxidative metabolism under physiological conditions. PMID- 12763072 TI - Deletion of the N-terminus of murine map2 by gene targeting disrupts hippocampal ca1 neuron architecture and alters contextual memory. AB - Microtubule-associated protein-2 (MAP2) is a brain specific A-kinase anchoring protein that targets the cyclic AMP-dependent protein kinase holoenzyme (PKA) to microtubules. Phosphorylation of MAP2 by different protein kinases is crucial for neuronal growth. The N-terminus of MAP2 contains the binding site for regulatory subunit II of cAMP-dependent protein kinase (PKA-RIIbeta). Using homologous recombination, we created a mutant line of mice (delta1-158) that express truncated MAP2 lacking the N-terminal peptide and the PKA binding site. Deletion of the PKA binding site from the MAP2 gene resulted in decreased efficiency of MAP2 phosphorylation. Biochemical and immunohistochemical studies demonstrate major changes in the morphology of hippocampal neurons in delta1-158 mice. Behavioral tests indicate that delta1-158 mice were impaired (exhibited less conditioned freezing) relative to Wild-Type (WT) controls during a test of contextual, but not during auditory cue, fear conditioning when tested at 8 weeks or 8 months of age. The delta1-158 mice displayed a heightened sensitivity to shock at 8 weeks, but not at 8 months of age. We conclude that PKA binding to MAP2 and MAP2 phosphorylation is essential for the selective development of contextual memory. PMID- 12763073 TI - The anabolic-androgenic steroid nandrolone induces alterations in the density of serotonergic 5HT1B and 5HT2 receptors in the male rat brain. AB - Anabolic-androgenic steroids (AAS) are partly misused by males in order to become brave and intoxicated and these agents are highly associated with psychosis, disinhibition, aggression and acts of violence. Since such behavioral states have been related to an imbalanced serotonergic system and the involvement of the serotonergic 5HT(1B) and the 5HT(2) receptors, it was important to discern the impact of AAS on these receptors. The objective of our study was to investigate the effects of 2 weeks of treatment with the AAS nandrolone decanoate at three different doses (1, 5, 15 mg/kg/day) on the total specific binding of the radioligands [(125)I]-(+/-)-1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI) (5HT(2) receptors) by autoradiography. All doses caused a significant down regulation of the 5HT(1B) receptor density in the hippocampal CA(1) and in the medial globus pallidus and a significant up-regulation of the 5HT(2) receptor density in the nucleus accumbens shell. Alterations in receptor density were also observed in the lateral globus pallidus, ventromedial hypothalamus, the amygdala and in the intermediate layers of various cortex regions. In conclusion, serotonergic 5HT(1B) or 5HT(2) receptors are likely to play important roles in mediating observed emotional states and behavioral changes among AAS abusers. PMID- 12763074 TI - Distribution of and organisation of dorsal horn neuronal cell bodies that possess the muscarinic m2 acetylcholine receptor. AB - Cholinergic systems in the dorsal horn are involved in antinociception but little is known about the organisation of receptors that mediate this process. In this study we examined immunocytochemical properties of dorsal horn neuronal cell bodies that express the m2 muscarinic acetylcholine receptor. Tissue was examined with confocal laser scanning microscopy and quantitative analysis performed. Immunoreactive cells were found throughout the dorsal horn and in lamina X. Quantitative analysis revealed that 22% of neuronal somata in the dorsal horn possess the receptor. The greatest concentration of cells was found in deeper laminae (IV-VI) and around lamina X. A proportion of cholinergic cells (labelled with an antibody against choline acetyltransferase) were immunoreactive for the receptor (approximately, 40% of dorsal horn cells and 44% of lamina X cells). Populations of presumed inhibitory interneurons also displayed immunoreactivity for the receptor. Between 27-34% of cells immunoreactive for GABA, nitric oxide synthase and the somatostatin receptor(2A) expressed the receptor but only 8% of parvalbumin-immunoreactive cells displayed receptor immunoreactivity. Cells labelled with neurotensin, which belong to a subgroup of excitatory neurons, displayed no receptor immunoreactivity. A small number neurokinin-1 receptor immunoreactive cells in lamina I possessed m2 immunoreactivity but 42% of laminae III/IV neurokinin-1 cells possessed it. This study shows that a significant proportion of cell bodies in the dorsal horn express the muscarinic m2 acetylcholine receptor. The receptor is present on some cholinergic neurons and therefore may function as an autoreceptor. It is associated with inhibitory local circuit neurons and may have a role in the modulation of specific inhibitory systems. It is also found on a proportion of projection cells that possess the neurokinin-1 receptor. This could be the basis of some of the antinociceptive actions of acetylcholine. PMID- 12763075 TI - Transduction of light in the suprachiasmatic nucleus: evidence for two different neurochemical cascades regulating the levels of Per1 mRNA and pineal melatonin. AB - The suprachiasmatic nucleus (SCN) contains a circadian clock and regulates melatonin synthesis in the pineal gland. Light exposure during the subjective night acutely increases the mRNA levels of the Period (Per)1 gene in the SCN and acutely suppresses melatonin levels in the pineal gland. Activation of N-methyl-D aspartate (NMDA) receptors in the SCN has been demonstrated to phase-shift the circadian clock in a manner similar to light. We tested the hypothesis that activation of excitatory amino acid (EAA) receptors in the SCN mediates the acute effects of light on Per1 mRNA levels and pineal melatonin. NMDA, injected into the SCN of Syrian hamsters during the night, acutely suppressed melatonin levels in the pineal gland. Both the NMDA receptor antagonist 2-amino-5 phosphonopentanoic acid (AP5) and the alpha-amino-3-hydroxy-5 methylisoxazoleproprionic acid (AMPA)/kainate receptor antagonist 6,7 dinitroquinoxaline-2,3-dione (DNQX) inhibited the light-induced increase of Per1 mRNA levels in the SCN. In the same animals, however, these antagonists had no effect on the ability of light to suppress pineal melatonin. These results support the hypothesis that EAA receptor activation in the SCN is necessary for the acute effects of light on Per1 mRNA levels. They also indicate that NMDA receptor activation in the SCN is sufficient but may not be necessary for the acute effects of light on pineal melatonin. These data suggest that there may be at least two different neurochemical cascades that transduce the effects of light in the SCN PMID- 12763077 TI - A thymulin analogue peptide with powerful inhibitory effects on pain of neurogenic origin. AB - The effects of a synthetic peptide analog of thymulin (PAT) were tested on nociceptive behavior in two animal models for peripheral mononeuropathy and in another two models for capsaicin-induced hyperalgesia. Treatment with PAT (0.25 25 microg/rat, i.p.) produced significant reduction of the mechanical allodynia and heat hyperalgesia in rats subjected to either chronic constriction injury (CCI) or spared nerve injury (SNI) models for mononeuropathy. Cold allodynia was moderately reduced in the CCI model. The inhibition of neuropathic manifestations peaked at 1-2 h post-treatment and disappeared in 3-4 h. Daily treatment with PAT, however, produced progressive attenuation of all neuropathic manifestations in the SNI model. On the other hand, pretreatment with similar doses of PAT produced dose-dependent reduction of the hyperalgesia induced by intraplantar injection of capsaicin (10 microg in 50 microl). The highest dose of PAT (50 microg) produced significant reduction of abdominal aversive behavior induced by i.p injection of capsaicin (20 microg in 100 microl). Compared with the effects of treatment with morphine or meloxicam (injected at single doses known to produce analgesia), PAT exerted equal or stronger inhibitory effects on neuropathic manifestations. The reported results suggest a possible direct action of PAT on afferent nerve fibers but its mechanisms remain to be determined. PMID- 12763076 TI - Adenosine suppresses the response of neurons to gaba in the superficial laminae of the rat spinal dorsal horn. AB - With the nystatin-perforated whole-cell patch-clamp recording technique, the modulatory effects of adenosine on GABA-activated whole-cell currents were investigated in neurons acutely dissociated from the superficial laminae (laminae I and II) of the rat spinal dorsal horn. The results showed that: (1) GABA acted on GABA(A) receptor and elicited inward Cl(-) currents (I(GABA)) at a holding potential (V(H)) of -40 mV; (2) adenosine suppressed GABA-induced Cl(-) current with affecting neither the reversal potential of I(GABA) nor the apparent affinity of GABA to its receptor; (3) N6-cyclo-hexyladenosine, a selective A(1) adenosine receptor agonist, mimicked the suppressing effect of adenosine on I(GABA), whereas 8-cyclopentyl-1,3-dipropylxanthine, a selective A(1) adenosine receptor antagonist, blocked the suppressing effect of adenosine; (4) chelerythrine, an inhibitor of protein kinase C, reduced the suppressing effect of adenosine on I(GABA); (5) pretreatment with 1,2-bis-(2-aminophenoxy) ethane N,N,N',N'-tetraacetic acid tetrakis (acetoxy-methyl) ester, a Ca(2+) chelator, did not affect adenosine-induced suppression of I(GABA). The results indicate that: (1) the suppression of adenosine on I(GABA) is mediated by adenosine A(1) receptor and through a Ca(2+)-independent protein kinase C transduction pathway; (2) the interactions between adenosine and GABA might be involved in the modulation of nociceptive information transmission at spinal cord level. PMID- 12763078 TI - The acquisition, retention and reversal of spatial learning in the morris water maze task following withdrawal from an escalating dosage schedule of amphetamine in wistar rats. AB - Two experiments were carried out to evaluate the effects of amphetamine withdrawal in rats on spatial learning in the water maze. A schedule of repeated d-amphetamine administration lasting for 6 days, with three injections per day (1 5 mg/kg, i.p.), was employed. Experiment 1 demonstrated that amphetamine withdrawal did not impair the acquisition of the water maze task (third to fourth withdrawal days), but amphetamine-withdrawn rats made more target-zone visits and reached the former location of the platform quicker than controls during the probe test (fifth withdrawal day). In experiment 2, retention of the location of the escape platform was assessed in animals having been pre-trained on the water maze task before treatment. On the third withdrawal day, retention of the former platform location was assessed in a probe test. Retention was only clearly seen in the measure of target zone visits, and performance did not differ between groups. Next, the animals were trained to escape to a new location in the water maze on withdrawal days 4-5. A reversal effect could be discerned across the first four trials, as evident by the animals' tendency to search in the former target quadrant. This interfered with the new learning, but amphetamine-withdrawn animals appeared to overcome it more rapidly than saline-treated controls. This finding is consistent with the view that amphetamine withdrawal can enhance behavioural switching, which could be expressed as a reduction of proactive interference during learning; and, it is in line with our previous finding that latent inhibition is also attenuated during amphetamine withdrawal. PMID- 12763079 TI - Voltage-dependent K+ currents in rat cardiac dorsal root ganglion neurons. AB - We have assessed the expression and kinetics of voltage-gated K(+) currents in cardiac dorsal root ganglion (DRG) neurons in rats. The neurons were labelled by prior injection of a fluorescent tracer into the pericardial sack. Ninety-nine neurons were labelled: 24% small (diameter<30 microm), 66% medium-sized (diameter 30 microm>.48 microm) and 10% large (>48 microm) neurons. Current recordings were performed in small and medium-sized neurons. The kinetic and pharmacological properties of K(+) currents recorded in these two groups of neurons were identical and the results obtained from these neurons were pooled. Three types of K(+) currents were identified:a) I(As), slowly activating and slowly time dependently inactivating current, with V(1/2) of activation -18 mV and current density at +30 mV equal to 164 pA/pF, V(1/2) of inactivation at -84 mV. b) I(Af) current, fast activating and fast time-dependently inactivating current, with V(1/2) of activation at two mV and current density at +30 mV equal to 180 pA/pF, V(1/2) of inactivation at -26 mV. At resting membrane potential I(As) was inactivated, whilst I(Af), available for activation. The I(As) currents recovered faster from inactivation than I(Af) current. 4-Aminopiridyne (4-AP) (10 mM) and tetraethylammonium (TEA) (100 mM) produced 98% and 92% reductions of I(Af) current, respectively and 27% and 66% of I(As) current, respectively. c) The I(K) current that did not inactivate over time. Its V(1/2) of activation was -11 mV and its current density equaled 67 pA/pF. This current was inhibited by 95% (100 mM) TEA, whilst 4-AP (10 mM) produced its 23% reduction. All three K(+) current components (I(As), I(Af) and I(K)) were present in every small and medium-sized cardiac DRG neuron. We suggest that at hyperpolarized membrane potentials the fast reactivating I(As) current limits the action potential firing rate of cardiac DRG neurons. At depolarised membrane potentials the I(Af) K(+) current, the reactivation of which is very slow, does not oppose the firing rate of cardiac DRG neurons. PMID- 12763080 TI - Photolysis of caged inositol 1,4,5-trisphosphate induces action potentials in frog vomeronasal microvillar receptor neurones. AB - To study the effect of inositol 1,4,5-trisphosphate (IP(3)) in isolated frog vomeronasal microvillar receptor neurones, whole-cell recordings were performed with 0.5 microM caged IP(3) dissolved in the pipette solution. IP(3) was released by photolysis of caged IP(3) initiated by a 0.8-ms ultraviolet flash from a xenon flash lamp 70 s after the start of dialysis of caged IP(3) into the cell. Flash illuminating the whole receptor neurone with caged IP(3) triggered action potentials when the current was clamped at zero and a series of transient inward currents of 12-55 pA at a holding potential of -70 mV. The average number of spikes during the first 40 s after release of IP(3) was 7.2+/-2.5 (n=6, mean+/ S.E.M.). The average maximum current and the total inward transport of charge during the first 40 s after photolysis of caged IP(3) were -24+/-8.0 pA and 1.7+/-0.8 pC, respectively (n=5, mean+/-S.E.M.). Inward membrane currents of 12 55 pA after release of IP(3) were not observed with 50 microM La(3+) in the bath. Notably, flash focused on the terminal vesicle also triggered action potentials. No action potentials were observed following flash focused on the soma or outside the dendrite. The average number of spikes during the first 40 s after release of IP(3) initiated by flash spatially restricted to the terminal vesicle was 5.0+/ 2.0 (n=4, mean+/-S.E.M.).The present study indicates that local release of IP(3) in the terminal vesicle of the vomeronasal neurones triggers transient depolarizations and induces action potentials. We suggest that IP(3) might be a second messenger in the vomeronasal microvillar receptor neurones. PMID- 12763081 TI - Contraction-sensitive skeletal muscle afferents inhibit arterial baroreceptor signalling in the nucleus of the solitary tract: role of intrinsic GABA interneurons. AB - Arterial baroreceptor and skeletal muscle receptor afferents relay sensory information to the nucleus of the solitary tract (NTS) during exercise. Previous studies have suggested that skeletal muscle afferent input inhibits baroreflex function; however, detailed information on the role of muscle afferents and GABAergic mechanisms in the NTS is limited. Furthermore, identification of specific afferent modalities that activate GABAergic neurons in the NTS remains unknown. In the present study, we examined the neuroanatomical and physiological interactions between spinal dorsal horn cells that transmit contraction-sensitive input from skeletal muscle and GABAergic interneurons in the NTS. Biotinylated dextran amine (BDA, 10%, 25-100 nL) microinjection into dorsal horn of the cervical spinal cord was combined with glutamate decarboxylase (GAD) immunohistochemistry to visualize the nature of the relationship of BDA-labeled fibers in the NTS with GAD immunoreactivity (GAD-ir). BDA-labeled axons and terminal processes were localized in the medial, commissural, dorsomedial and dorsolateral subdivisions of the caudal NTS. Moreover, BDA-labeled fibers were observed in close proximity to GAD-ir structures throughout these regions of the NTS. The physiological interaction between skeletal muscle receptor and arterial baroreceptor afferents was investigated using an arterially perfused, decerebrate rat preparation. Activation of skeletal muscle afferents by electrically evoked twitch contraction of the forelimb attenuated baroreflex responsiveness (BR, calculated as the ratio of changes in heart rate to systemic pressure) from 1.5+/-0.3 bpm.mm Hg(-1) to -0.1+/-0.1 bpm.mm Hg(-1) (control versus contraction, P<0.05, n=15). However, forelimb contraction failed to inhibit the reflex bradycardia evoked by activation of peripheral chemoreceptor afferents, indicating a reflex-specific action. Bilateral microinjection of bicuculline methiodide (BIC, 10 microM, 40-60 nL) into the caudal NTS restored baroreflex responsiveness during contraction (-1.6+/-0.2 versus -0.1+/-0.1 versus -1.5+/-0.2 bpm.mmHg(-1), control versus contraction versus contraction+BIC P<0.05, n=8). We conclude that activation of ascending spinal neurons from the cervical dorsal horn by contraction-sensitive skeletal muscle afferents selectively inhibits arterial baroreceptor signaling in the NTS via activation of a GABAergic mechanism. PMID- 12763082 TI - Fasting is a physiological stimulus of vagus-mediated enhancement of nociception in the female rat. AB - The vagus nerve modulates nociception by a mechanism dependent upon gonadal hormones and the adrenal medulla. In the present study we tested the hypothesis that this modulation is dynamically controlled by physiological stimulation of structures innervated by the subdiaphragmatic vagus. Specifically, food deprivation (fasting) was employed to increase activity in the subdiaphragmatic vagus, and the experiments were performed mainly in female rats because our previous observations suggested that baseline activity in the pathway is lower in females than in males. Consistent with the hypothesis, after a 48-h fast, female rats exhibited increased nociceptive behavior in the formalin test. In contrast, fasting had no effect on formalin-evoked nociceptive behavior in male rats. The fasting-induced effect on nociception appears to be mediated by the vagus nerve since it is prevented by subdiaphragmatic vagotomy. Also similar to the previously characterized vagus-mediated modulation, the effect of fasting in the female is blocked by gonadectomy or adrenal medullectomy, and hormone replacement with 17beta-estradiol in gonadectomized female rats restored the effect of fasting. Decreased glucose metabolism apparently does not play a significant role in the effect of fasting on nociception, since the effect was unchanged when 5% glucose was provided in the drinking water throughout the fasting period. On the other hand, increasing the bulk content of the stomach (without providing nutrients) by infusion of petrolatum significantly attenuated the effect of fasting during the interphase period of the formalin response, suggesting that decreased gut distention, and possibly motility, are important in fasting-induced enhancement of nociception. These results indicate that fasting is a physiological activator of the vagus-mediated pain modulation pathway. This suggests the possibility that, especially in females, natural periodic changes in gut distention and motility may control an ongoing vagus-mediated adjustment in the organism's nociceptive sensitivity. PMID- 12763083 TI - Intrathecal substance p-saporin attenuates operant escape from nociceptive thermal stimuli. AB - Destruction of neurons in the superficial dorsal horn that express substance P receptor (NK-1R) has been reported to block development of behavioral hypersensitivity following peripheral sensitization of nociceptors. Baseline sensitivity was not altered in these rat models that assessed innate reflex responses (i.e. hind-paw withdrawal to thermal or mechanical stimulation). In the present study, we evaluated effects of intrathecal substance P-saporin (SP-sap), a toxin selective for cells expressing NK-1R, on operant escape responses of rats to thermal stimulation. For comparison, lick/guard reflex testing was performed. Injection of a modest dose (175 ng) of SP-sap into the lumbar subarachnoid space produced a partial loss of lamina I/II NK-1R-expressing dorsal horn neurons but did not affect NK-1R-expressing neurons in deeper laminae. Lick/guard responses to 0.3, 44 or 47 degrees C were not affected after SP-sap treatment, but escape responses to these temperatures were significantly attenuated. Three hours after application of mustard oil to the dorsal surface of both hind paws, escape from 44 degrees C was enhanced for controls but not SP-sap-treated rats. Lick/guard responses were enhanced by mustard oil for both SP-sap and control animals. Administration of morphine (1.0 mg/kg, s.c.) before testing decreased escape responding at 47 degrees C for both controls and SP-sap rats. Thus, partial loss of NK-1R-expressing neurons in the superficial dorsal horn attenuated thermal nociceptive sensitivity and prevented secondary hyperalgesia when studied with an operant algesia assay, in contrast to innate reflexes which were less sensitive to modification by intrathecal SP-sap. PMID- 12763084 TI - Dopamine depletion of the nucleus accumbens reverses isolation-induced deficits in prepulse inhibition in rats. AB - Rearing rats in social isolation from weaning into adulthood leads to deficits in prepulse inhibition and alterations in monoamine systems that modulate prepulse inhibition. For example, rats reared in social isolation have elevated dopamine levels in the nucleus accumbens. Previous studies in rats have shown that nucleus accumbens dopamine depletion with 6-hydroxydopamine blocks the prepulse inhibition-disruptive effects of amphetamine, an indirect dopamine agonist. We tested the hypothesis that prepulse-inhibition deficits in isolation-reared rats are dependent on elevated dopamine levels in the nucleus accumbens. Specifically, we examined whether nucleus accumbens dopamine depletion would attenuate the isolation-induced disruption of prepulse inhibition. Isolation-housed female Long Evans rats exhibited deficient prepulse inhibition. At 9 weeks post weaning, bilateral injections of 6-hydroxydopamine (8 microg/side) or ascorbic acid vehicle (0.1%) into the nucleus accumbens of social and isolation-reared rats were performed (8-10 rats per group). One week after surgery, prepulse inhibition deficits were exhibited by isolation-reared rats that received vehicle infusion into the nucleus accumbens, but not by those that received 6-hydroxydopamine infusions into the nucleus accumbens. 6-Hydroxydopamine infusions did not significantly change prepulse inhibition in socially reared rats. Behavioral and neurochemical evidence of nucleus accumbens dopamine depletion included: 1) a blockade of amphetamine-stimulated locomotor activity in nucleus accumbens 6 hydroxydopamine-infused isolated and socially reared rats; and 2) high performance liquid chromatography measurements demonstrating a significant depletion of accumbens dopamine and its major metabolites, in addition to decreases in dopamine, homovanillic acid, and 3,4-dihydroxyphenylacetic acid levels in the frontal cortex and anterior caudate. These data indicate that dopamine in the nucleus accumbens plays an essential role in the prepulse inhibition deficits associated with isolation rearing in female Long-Evans rats. The implication of a central role of nucleus accumbens dopamine in prepulse inhibition deficits in an animal model provides further evidence for a link between overactive dopamine function and sensorimotor-gating deficits in patients with schizophrenia. PMID- 12763085 TI - Blockade of ventral pallidal opioid receptors induces a conditioned place aversion and attenuates acquisition of cocaine place preference in the rat. AB - Peripheral administration of naloxone is known to produce a conditioned place aversion and to block cocaine-induced conditioned place preference. The ventral pallidum receives a dense enkephalinergic projection from the nucleus accumbens and is implicated as a locus mediating the rewarding and reinforcing effects of psychostimulant and opiate drugs. We sought to provide evidence for the involvement of pallidal opioid receptors in modulating affective state using the place-conditioning paradigm. Microinjection of naloxone (0.01-10 microg) into the ventral pallidum once a day for 3 days dose-dependently produced a conditioned place aversion when tested in the drug-free state 24 h after the last naloxone injection. This effect was reproduced using the mu-opioid receptor selective agonist D-Phe-Cys-Tyr-D-Trp-Orn-Thr-Pen-Thr-NH(2) (CTOP, 1 microg). Locomotor activity was reduced following injection of the highest dose of naloxone (10 microg) but elevated following CTOP (1 microg). Daily injection of cocaine (10 mg/kg) for 3 days produced a conditioned place preference 24 h later. This effect of cocaine was attenuated by concomitant intra-ventral pallidal injection of naloxone at a dose (0.01 microg) that had no significant aversive property when injected alone. In contrast, the locomotor activation induced by peripheral cocaine injection was unaffected by naloxone injection into the ventral pallidum. The data implicate endogenous opioid peptide systems within the ventral pallidum as regulators of hedonic status. PMID- 12763086 TI - Monoaminergic changes associated with socially induced sex reversal in the saddleback wrasse. AB - The process of sex reversal in fishes is socially mediated and requires a total reorganization of the hypothalamo-pituitary-gonadal axis. When the ratio of males to females in a population of saddleback wrasse (Thalassoma dupperrey) is too low, the largest female becomes male over the course of 6 to 8 weeks. This event requires the conversion of external social cues into internal chemical cues. In an attempt to investigate the role monoamines might play in this process, two females were housed together in floating enclosures in order to induce sex reversal in the larger. Brains were sampled at various time points throughout the process of sex reversal. Monoamines were measured in the amygdala, preoptic area, ventral hypothalamus, locus coeruleus and raphe nucleus. Changes were demonstrated in monoamine metabolism for all brain regions examined. The most important changes in monoamine-system activation were seen during the first week of sex reversal. It is during this time that transitional animals undergo behavioral sex reversal. There is an increase in serotonergic activity in the amygdala which is likely related to territorial acquisition. The absence of male aggression results in a less stressful environment for the female and a reduction in serotonergic activity in the preoptic area allowing for an increase in noradrenergic activity potentially triggering the reorganization of the reproductive axis. In the ventral hypothalamus, there is a decrease in noradrenergic and increase in dopaminergic activity associated with this change from female to male. The locus coeruleus shows an increase in noradrenergic activity later in the process of sex reversal which is probably a response to more circulating androgens. In the raphe nucleus, there is a decrease in serotonergic activity at the time of behavioral sex reversal. This decrease in serotonergic activity is linked to the behavioral component of sex reversal. This study suggests that monoamines play a very important role in both behavioral and gonadal sex reversal in the saddleback wrasse, the former under the control of serotonin in the raphe and the latter mediated via serotonergic effects on norepinephrine in the preoptic area. PMID- 12763087 TI - Excitatory GABA input directly drives seizure-like rhythmic synchronization in mature hippocampal CA1 pyramidal cells. AB - GABA, which generally mediates inhibitory synaptic transmissions, occasionally acts as an excitatory transmitter through intense GABA(A) receptor activation even in adult animals. The excitatory effect results from alterations in the gradients of chloride, bicarbonate, and potassium ions, but its functional role still remains a mystery. Here we show that such GABAergic excitation participates in the expression of seizure-like rhythmic synchronization (afterdischarge) in the mature hippocampal CA1 region. Seizure-like afterdischarge was induced by high-frequency synaptic stimulation in the rat hippocampal CA1-isolated slice preparations. The hippocampal afterdischarge was completely blocked by selective antagonists of ionotropic glutamate receptors or of GABA(A) receptor, and also by gap-junction inhibitors. In the CA1 pyramidal cells, oscillatory depolarizing responses during the afterdischarge were largely dependent on chloride conductance, and their reversal potentials (average -38 mV) were very close to those of exogenously applied GABAergic responses. Moreover, intracellular loading of the GABA(A) receptor blocker fluoride abolished the oscillatory responses in the pyramidal cells. Finally, the GABAergic excitation-driven afterdischarge has not been inducible until the second postnatal week. Thus, excitatory GABAergic transmission seems to play an active functional role in the generation of adult hippocampal afterdischarge, in cooperation with glutamatergic transmissions and possible gap junctional communications. Our findings may elucidate the cellular mechanism of neuronal synchronization during seizure activity in temporal lobe epilepsy. PMID- 12763088 TI - Spontaneous field potentials influence the activity of neocortical neurons during paroxysmal activities in vivo. AB - Field-potential recordings with macroelectrodes, and extra- and intracellular potentials with micropipettes were used to determine the influence of spontaneous field potentials on the activity of neocortical neurons during seizures. In vivo experiments were carried out in cats under anesthesia. Strong negative field fluctuations of up to 20 mV were associated with electroencephalogram "spikes" during spontaneously occurring paroxysmal activities. During paroxysmal events, action potentials displayed an unexpected behavior: a more hyperpolarized firing threshold and smaller amplitude than during normal activity, as determined with intracellular recordings referenced to a distant ground. Considering the transmembrane potential (the difference between extra- and intracellular potential) qualified this observation: firing threshold determined from the transmembrane potential did not decrease, and smaller action-potential amplitude was associated with depolarized firing threshold. The hyperpolarization of intracellular firing threshold was thus related to the field potentials. Similar field-potential effects on neuronal activities were observed when the paroxysmal events included very fast oscillations or ripples (80-200 Hz) that represent rapid fluctuations of field potentials (up to 5 mV in <5 ms). Neuronal firing was phase-locked to those oscillations. These results demonstrate that: (a) strong spontaneous field potentials influence neuronal behavior, and thus play an active role during paroxysmal activities; and (b) transmembrane potentials have to be used to accurately describe the behavior of neurons in conditions in which field potentials fluctuate strongly. Since neuronal activity is presumably the main generator of field potentials, and in return these potentials may increase neuronal excitability, we propose that this constitutes a positive feedback loop that is involved in the development and spread of paroxysmal activities, and that a similar feedback loop is involved in the generation of neocortical ripples. We propose a mechanism for seizure initiation involving these phenomena. PMID- 12763089 TI - Basal ganglia efferents to the brainstem centers controlling postural muscle tone and locomotion: a new concept for understanding motor disorders in basal ganglia dysfunction. AB - The present study is designed to elucidate how basal ganglia afferents from the substantia nigra pars reticulata (SNr) to the mesopontine tegmental area of the brainstem contribute to gait control and muscle-tone regulation. We used unanesthetized and acutely decerebrated cats (n=27) in which the striatum, thalamus and cerebral cortex were removed but the SNr was preserved. Repetitive stimulation (50 Hz, 10-60 microA, for 5-20 s) applied to a mesencephalic locomotor region (MLR), which corresponded to the cuneiform nucleus, and adjacent areas, evoked locomotor movements. On the other hand, stimulation of a muscle tone inhibitory region in the pedunculopontine tegmental nucleus (PPN) suppressed postural muscle tone. An injection of either glutamatergic agonists (N-methyl-D aspartic acid and kainic acid) or GABA antagonists (bicuculline and picrotoxin) into the MLR and PPN also induced locomotion and muscle-tone suppression, respectively. Repetitive electrical stimuli (50-100 Hz, 20-60 microA for 5-20 s) delivered to the SNr alone did not alter muscular activity. However stimulating the lateral part of the SNr attenuated and blocked PPN-induced muscle-tone suppression. Moreover, weaker stimulation of the medial part of the SNr reduced the number of step cycles and disturbed the rhythmic alternation of limb movements of MLR-induced locomotion. The onset of locomotion was delayed as the stimulus intensity was increased. At a higher strength SNr stimulation abolished the locomotion. An injection of bicuculline into either the PPN or the MLR diminished the SNr effects noted above. These results suggest that locomotion and postural muscle tone are subject to modulation by GABAergic nigrotegmental projections which have a partial functional topography: a lateral and medial SNr, for regulation of postural muscle tone and locomotion, respectively. We conclude that disorders of the basal ganglia may include dysfunction of the nigrotegmental (basal ganglia-brainstem) systems, which consequently leads to the production of abnormal muscle tone and gait disturbance. PMID- 12763090 TI - Neuroanatomical relationship between type 1 cannabinoid receptors and dopaminergic systems in the rat basal ganglia. AB - Dopamine and endocannabinoids are neurotransmitters known to play a role in the activity of the basal ganglia motor circuit. While a number of studies have demonstrated functional interactions between type 1 cannabinoid (CB1) receptors and dopaminergic systems, we still lack detailed neuroanatomical evidence to explain their relationship. Single- and double-labeling methods (in situ hybridization and immunohistochemistry) were employed to determine both the expression and localization of CB1 receptors and tyrosine hydroxylase (TH) in the basal ganglia. In the striatum, we found an intense signal for CB1 receptor transcripts but low signal for CB1 receptor protein, whereas in the globus pallidus and substantia nigra we found the opposite; no hybridization signal but intense immunoreactivity. Consequently, CB1 receptors are synthesized in the striatum and mostly transported to its target areas. No co-expression or co localization of CB1 receptors and TH was found. In the caudate-putamen, globus pallidus and substantia nigra, TH-immunoreactive fibers were interwoven with the CB1 receptor-immunoreactive neuropil and fibers. Our data suggest that the majority of the striatal CB1 receptors are located presynaptically on inhibitory GABAergic terminals, in a position to modulate neurotransmitter release and influence the activity of substantia nigra dopaminergic neurons. In turn, afferent dopaminergic fibers from the substantia nigra innervate CB1 receptor expressing striatal neurons that are known to also express dopamine receptors. In conclusion, these data provide a neuroanatomical basis to explain functional interactions between endocannabinoid and dopaminergic systems in the basal ganglia. PMID- 12763091 TI - Role of metabotropic glutamate receptor subtype 5 (mGluR5) in the maintenance of cold hypersensitivity following a peripheral mononeuropathy in the rat. AB - The present series of experiments were designed to examine the contribution of metabotropic glutamate receptor subtype 5 (mGluR5) to neuropathic pain by determining the effects of the selective mGluR5 antagonist MPEP (2-methyl-6 (phenylethynyl)-pyridine) on neuropathy-induced cold hypersensitivity. Unilateral chronic constriction injury (CCI) to the sciatic nerve in rats produced an increase in the number of hind paw withdrawals from a cold surface (4 +/- 2 degrees C) which was dose-dependently inhibited by systemic (i.p.) injection of MPEP (ID(50) = 11.3 mg/kg). In vivo brain mGluR5 receptor occupancy following systemic (i.p.) MPEP revealed that >90% occupancy is required for behavioral efficacy. Intracerebroventricular (i.c.v.) injection of MPEP dose-dependently inhibited CCI-induced cold hypersensitivity (ID(50) = 123.5 nmol), while microinjection of MPEP directly into the rostral ventromedial medulla (RVM) potently inhibited this hypersensitivity (ID(50) = 1.3 pmol). A role for mGluR5 in the RVM was further supported by the observation that intra-RVM injection of the mGluR5 agonist CHPG (10 nmol; 2-chloro-5-hydroxyphenylglycine) produced cold hypersensitivity in naive rats that was blocked by pretreatment with intra-RVM MPEP (3 nmol). Intrathecal (500 nmol; i.t.) or intraplantar (300 nmol; i.pl.) injection of MPEP was ineffective in reversing CCI-induced cold hypersensitivity. These results demonstrate that mGluR5 contributes to cold hypersensitivity following peripheral neuropathy exclusively at supraspinal sites in the CNS. Additionally, mGluR5 in the RVM significantly contributes to the maintenance of cold hypersensitivity, likely via activation of descending nociceptive facilitatory systems. PMID- 12763092 TI - The pharmacology of spontaneously open alpha 1 beta 3 epsilon GABA A receptor ionophores. AB - Human alpha(1)beta(3) epsilon GABA(A) receptors were expressed in Xenopus oocytes and examined using the conventional two-electrode voltage-clamp technique and compared to alpha(1)beta(3)gamma(2) receptors. The effects of several GABA(A) agonists were studied, and the allosteric modulation of the channel by a number of GABAergic modulators investigated. The presence of the epsilon subunit increased the potency and efficacy of direct activation by partial GABA(A) agonists (piperidine-4-sulphonic acid and thio-4-PIOL), pentobarbital and neuro steroids. Direct activation by 3-hydroxylated neurosteroids was restricted to 3alpha epimers, while chirality at C5 was indifferent. The 3beta-sulfate esters of pregnenolone and dehydroepiandrosterone inhibited the spontaneous currents with efficacies higher, while bicuculline methiodide and SR 95531 did so lower than picrotoxin and TBPS. Furosemide, fipronil, triphenylcyanoborate and Zn(2+) blocked the spontaneous currents of alpha(1)beta(3) epsilon receptors with different efficacies. Flunitrazepam and 4'-chlorodiazepam inhibited the spontaneous currents with micromolar potencies. In conclusion, spontaneously active alpha(1)beta(3) epsilon GABA(A) receptors can be potentiated and blocked by GABAergic agents within a broad range of efficacy. PMID- 12763093 TI - Effects of gamma2S subunit incorporation on GABAA receptor macroscopic kinetics. AB - GABA(A) receptors, the major inhibitory neurotransmitter receptors in the mammalian central nervous system, are heteropentameric proteins. We are interested in understanding the contribution of the gamma subunit to the kinetic properties of GABA(A) receptors. Studies in Xenopus oocytes have suggested that co-expression of alpha1, beta2, and gamma 2S subunits results in the formation of both alpha beta and alpha betagamma receptors (Boileau et al. 2002a; Boileau et al., 1998). Here, we have used an excess of the gamma 2S subunit in transfections of HEK293 cells to bias expression toward alpha beta gamma-containing receptors. Using rapid application and whole cell patch clamp techniques, we found that incorporation of the gamma subunit eliminated the rapid phases of desensitization and accelerated deactivation, consistent with a proposed role of desensitization in slowing deactivation. In addition, alpha betagamma receptors had an increased GABA EC(50), reduced sensitivity to block by Zn(2+), and did not display outward rectification as compared to alpha beta receptors. PMID- 12763094 TI - Chronic AMPA receptor potentiator (LY451646) treatment increases cell proliferation in adult rat hippocampus. AB - Stress-induced neuronal atrophy and death in the hippocampus may play an important role in the etiology of clinical depression. Conventional antidepressants can stimulate hippocampal neurogenesis after chronic administration. AMPA receptor potentiators (ARPs) such as LY392098 and LY451616 are active in both the forced swim test and the tail suspension test, two behavioral despair procedures widely used to predict antidepressant efficacy. Unlike traditional antidepressants, this group of compounds does not affect extracellular concentrations of biogenic amines. In this study, we investigated the effect of LY451646 on progenitor cell proliferation in adult rat hippocampus. Male Sprague-Dawley rats (n = 4-5 per group) received either single or chronic (21 days) doses of LY451646 (0.025-0.5 mg/kg). Bromodeoxyuridine (BrdU) injections and immunohistochemistry were performed 30 min and 24 h after the last drug injection, respectively. Results show that chronic LY451646 treatment increased progenitor cell proliferation (approximately 45%) in the dentate gyrus in a dose-dependent manner. This upregulation of BrdU labeling appeared as an increase in the number of cells arranged in clusters. Similarly, a significant increase in the number of cells in clusters was observed after a single injection of LY451646 (0.05 mg/kg), although the increase in total number of BrdU-positive cells (approximately 30%) did not reach statistical significance. This is the first in vivo study showing the modulation of progenitor cell proliferation by an ARP. These findings suggest that the antidepressant-like activity of ARPs in animals may be attributed, at least in part, to the regulation of progenitor cell proliferation in the hippocampus. PMID- 12763095 TI - A distinct distribution of functional presynaptic 5-HT receptor subtypes on GABAergic nerve terminals projecting to single hippocampal CA1 pyramidal neurons. AB - 5-HT is known to modify the excitability of GABAergic interneurons projecting to hippocampal CA1 neurons. In this study we investigate the presence and functionally characterize the 5-HT receptor subtypes found on the presynaptic nerve terminals of these GABAergic neurons. Using conventional whole-cell patch recording, we confirmed that the 5-HT(1A) agonist, 8-hydroxy-2 dipropylaminotetralin, presynaptically decreased electrically evoked GABA release while the 5-HT(3) agonist, m-chlorophenylbiguanide (mCPBG), presynaptically facilitated release. Using the 'synaptic bouton preparation', where CA1 neurons are acutely isolated with functional nerve terminals/boutons remaining adherent, we next showed that these receptor subtypes are found presynaptically. We next used the technique of focal stimulation of a single bouton in this preparation to further investigate the distribution of these 5-HT receptor subtypes. We found that all boutons contained inhibitory 5-HT(1A) receptors while a subset of boutons showed both 5-HT(1A) and excitatory 5-HT(3) receptors. No boutons were detected which contained only 5-HT(3) receptors. Our studies show that presynaptic 5-HT receptor subtypes are found presynaptically and are not uniformly distributed. This provides another potential mechanism whereby 5-HT can modulate GABA release and hence the excitability of hippocampal neurons. PMID- 12763096 TI - The hypothermic effect of 5-CT in mice is mediated through the 5-HT7 receptor. AB - The 5-HT(7) receptor is a recent addition to the 5-HT receptor family and to date there is no clear idea as to its potential role in the CNS. The receptor has been mapped by in situ hybridization and 5-HT(7)-like immunoreactivity and has been detected in discrete areas of the brain including the hypothalamus (Oliver et al., 1999). This suggests the receptor may be involved in temperature regulation and have shown that a selective 5-HT(7) receptor antagonist reverses the hypothermic effect of 5-CT in guinea-pigs. The current study confirmed that the 5 HT(7) receptor antagonists, SB-269970 (1-30 mg/kg, i.p.) and SB-258719 (5-20 mg/kg, i.p.), but not the 5-HT(1A) receptor antagonist, WAY 100635(0.1-1 mg/kg, s.c.), or the 5-HT(1B/D) antagonist, GR127935 (1.25-5 mg/kg, i.p.), reversed the hypothermic effect of 5-CT in mice. In addition the effect of 5-CT on body temperature was examined on 5-HT(7) receptor null mutant mice. 5-CT (0.1-1 mg/kg, i.p.) significantly reduced rectal temperature in wildtype but not 5-HT(7) receptor knockout mice. This suggests that the hypothermic effects of 5-CT are mediated through the 5-HT(7) receptor. All procedures were carried out in accordance with the UK Animals (Scientific Procedures) Act (1986). PMID- 12763097 TI - Pharmacological and functional characterisation of dopamine D4 receptors in the rat retina. AB - In the retina, activation of dopamine receptors, particularly the D2-like family (D2, D3, D4 receptor subtypes), with quinpirole suppresses the light sensitive cAMP pool and inhibits melatonin synthesis in photoreceptor cells. We have characterised rat retinal D4 receptors using the D4 selective radioligand [(125)I] L-750667 which bound specifically and saturably to rat retinal membranes with high affinity (K(d) 0.06+/-0.02 nM) and exhibited a D4 receptor pharmacology. Comparison of the binding kinetics of [(125)I] L-750667 and [(3)H] spiperone revealed B(max) values of 134+/-27 fmol/mg and 219+/-47 fmol/mg respectively, indicating that the dopamine D4 receptor is a major component of D2 like dopamine receptors in the rat retina. Modulation of retinal cAMP levels by quinpirole was used to evaluate the functional relevance of rat retinal dopamine D4 receptors. Quinpirole (0.03-3 micro ) produced a dose-related decrease of the light sensitive cAMP pool which was reversed by haloperidol, clozapine and the D4 selective antagonist, L-745870 with a rank order of potency suggesting that the quinpirole effect is due to activation of the dopamine D4 receptors. The D2 selective ligand L-741626 had no effect on the quinpirole response confirming that the D4 receptor is the major receptor subtype mediating dopamine induced suppression of adenylate cyclase in the retina. PMID- 12763098 TI - Evidence for regulation of body temperature in rats by dopamine D2 receptor and possible influence of D1 but not D3 and D4 receptors. AB - The dopamine D(3) receptor agonist PD 128907 decreased body temperature in the rat. The selective dopamine D(3) and D(4) receptor antagonists, A-437203 and L 745,870, respectively, did not prevent this effect. In contrast, PD 128907 induced hypothermia was antagonized by SCH 23390, a selective D(1) receptor antagonist, and by L-741,626, a selective D(2) receptor antagonist. Moreover, the selective D(2) receptor agonist trihydroxy-N-n-propylnoraporphine (TNPA) elicited a robust hypothermia which was prevented by pretreatment with L-741,626 but not by A-437203. In agreement with previous data obtained in D(3) knock-out mice, present results suggest that D(2) rather than D(3) receptors mediate dopamine receptor agonist-induced hypothermia in rats. In addition, it appears that both D(1) and D(2) receptors may be involved in a cooperative manner. PMID- 12763099 TI - Attentional effects of nicotinic agonists in rats. AB - Nicotine can increase stimulus detection, response rate and speed in the five choice serial reaction time task, a rodent test of attention. In the present experiments, four other nicotinic agonists with different pharmacological profiles were compared in the same procedure. The response profile of epibatidine resembled that previously obtained with nicotine in that response accuracy was enhanced and omission errors and correct response latency decreased. ABT-418 transiently increased accuracy in the first 10 min of test sessions and reduced response latency. Isoarecolone caused a dose-related increase in accuracy, but had no effect on omissions or response latency. This absence of effects on response rate- or speed-related measures may be related to its previously reported reduced ability to release dopamine as compared with nicotine. The alpha7-agonist AR-R17779 was without effect on any measure, indicating that this receptor subtype may not mediate nicotinic effects on attention. Affinity constants of compounds, determined in competition binding assays targeting the alpha4beta2, alpha7, alpha3beta4 and alpha3beta2* nAChR subtypes, could not explain the differential behavioural effects observed. Differences in their functional efficacy at nAChR subtypes may instead be responsible. The finding that attentional performance and response rate and speed can be selectively modulated by nicotinic agonists is encouraging for the development of drugs with therapeutic properties similar to those of nicotine but with reduced unwanted effects. PMID- 12763100 TI - Administration of caspase 3 inhibitor during and after status epilepticus in rat: effect on neuronal damage and epileptogenesis. AB - Symptomatic temporal lobe epilepsy typically develops in three phases: brain damage --> epileptogenesis --> spontaneous seizures (epilepsy). The challenge is to prevent epileptogenesis after injury. We hypothesized that alleviation of damage by caspase inhibitors will reduce epileptogenesis or at least have disease modifying effects (less severe epilepsy, milder cognitive decline). Epileptogenesis was triggered by amygdala stimulation-induced status epilepticus (SE) in rats and spontaneous seizures were monitored with video electroencephalography (EEG). First, we tested the neuroprotective effect of a 1 week treatment with caspase 1, 3 or 9 inhibitors (3 micro g/d/i.c.v., started 3 h after the beginning of SE). The least damage to the hippocampus was observed in animals treated with the caspase 3 inhibitor (z-DEVD-fmk) which reduced the enzyme activity to 6% of that in the vehicle group. Thus, z-DEVD-fmk was chosen for long-term studies, in which the treatment regime remained the same except the dose was doubled (6 micro g/d/i.c.v.). Video-EEG monitoring was performed for 3 to 4 weeks, starting either 8 or 14 weeks after SE. One group of animals was tested in water-maze and fear-conditioning tests, and all animals were perfused for histological analysis. Treatment with the caspase 3 inhibitor neither prevented the development of epilepsy, nor had any disease-modifying effects. Mossy fibre sprouting, however, was reduced. The present data indicate that administration of z-DEVD-fmk monotherapy was not antiepileptogenic despite its short-term neuroprotective effects. These findings challenge the idea that prevention of cell death is the primary target for the development of antiepileptogenic compounds. PMID- 12763101 TI - Regulation of contextual fear conditioning by baseline and inducible septo hippocampal cyclin-dependent kinase 5. AB - In this work, we confirm the novel role of cyclin-dependent kinase (Cdk) 5 in associative learning by demonstrating that injection of the Cdk5 inhibitor butyrolactone I into the lateral septum or hippocampus profoundly impaired context-dependent fear conditioning of C57BL/6J mice. However, unlike the inducible up-regulation of Cdk5 and its regulator p35 observed in Balb/c mice, high baseline levels, which were not affected by fear conditioning, were found in C57BL/6J mice. Surprisingly, microinjections of butyrolactone I into the lateral septum or hippocampus significantly decreased baseline Cdk5 activity within the entire septo-hippocampal circuitry, suggesting a functional link between septal and hippocampal Cdk5 activity. Significantly higher levels of the transcription factor Sp4 in the septo-hippocampal system of C57BL/6J mice may account for the high baseline Cdk5/p35 production. On the other hand, the stronger cFos production observed in the lateral septum of fear conditioned Balb/c mice may be responsible, at least in part, for the inducible up-regulation of Cdk5 in this strain. These results suggest that the role of Cdk5 in memory consolidation is strain independent and functionally related to the septo-hippocampal circuitry. However, the molecular regulation of baseline and inducible Cdk5 protein might be different among individual mouse strains and possibly other species. PMID- 12763102 TI - Quercetin, a bioflavonoid, attenuates haloperidol-induced orofacial dyskinesia. AB - Chronic treatment with neuroleptics leads to the development of abnormal orofacial movements described as vacuous chewing movements (VCMs) in rats. Vacuous chewing movements in rodents are widely accepted as one of the animal models of tardive dyskinesia. Oxidative stress and the products of lipid peroxidation are implicated in the pathophysiology of various neurological disorders including tardive dyskinesia. In the present study chronic haloperidol (1.0 mg kg(-1) for 21 days) treatment induced vacuous chewing movements and tongue protrusions in rats. Co-administration of quercetin, a bioflavonoid, dose dependently (25-100 mg kg(-1)) reduced haloperidol-induced vacuous chewing movements and tongue protrusions. Biochemical analysis revealed that chronic haloperidol treatment induces lipid peroxidation and decreases the glutathione (GSH) levels in the forebrains of rats. The antioxidant defense enzymes, superoxide dismutase (SOD) and catalase were also decreased due to chronic haloperidol treatment. Co-administration of quercetin (25-100 mg kg(-1)) significantly reduced the lipid peroxidation and restored the decreased glutathione levels in these animals. Further quercetin (50-100 mg kg(-1)) also reversed the haloperidol-induced decrease in forebrain SOD and catalase levels in rats. The major findings of the present study suggested that oxidative stress plays a significant role in neuroleptic-induced orofacial dyskinesia and quercetin co-administration reverses these behavioral and biochemical changes. Quercetin, a naturally occurring bioflavonoid could prove to be a useful agent in neuroleptic-induced orofacial dyskinesia. PMID- 12763103 TI - Block of slow axonal transport and axonal growth by brefeldin A in compartmented cultures of rat sympathetic neurons. AB - Disruption of the Golgi by brefeldin A (BFA) has been reported to block fast axonal transport and axonal growth. We used compartmented cultures of rat sympathetic neurons to investigate its effects on slow axonal transport. BFA (1 micro g/ml) applied to cell bodies/proximal axons for 6-20 h disrupted the Golgi, reversibly blocked axonal growth, and reversibly blocked anterograde transport of all proteins, including tubulin. The retrograde transport of nerve growth factor (NGF) was also blocked. The phosphorylation of Erk1 and Erk2 in response to NGF was unaffected after 6 h of treatment with BFA, suggesting that the block of axonal transport was specific and direct. Consistent with its principal site of action at the Golgi, no effects were observed when BFA was applied only to the distal axons. Block of fast anterograde and retrograde axonal transport is consistent with the role of the Golgi in supplying transport vesicles. Block of slow axonal transport was surprising, and further results indicated that transport of tubulin en route along the axon was arrested by application of BFA to the cell bodies, suggesting that a continuous supply of anterograde transport vesicles from the Golgi is required to maintain slow axonal transport of cytoskeletal proteins. PMID- 12763104 TI - Defining the problem: the epidemiology of preterm birth. AB - Preterm birth is the major clinical problem associated with perinatal mortality, serious neonatal morbidity and moderate to severe childhood disability in prosperous countries. Its prevalence is affected by the way in which gestational age is assessed, by national differences in the registration of births, associated practices, such as burial costs, or maternity benefits, which encourage or discourage registration, and by the perceived viability of extremely preterm infants. Despite these uncertainties, there is reliable evidence that preterm births are increasing, especially births before 28 weeks gestation. Contributing factors include births following assisted reproductive therapy and ovulation induction, especially multiple births, and the increasing proportion of births among women >34 years. On the other hand, improvements in neonatal care have substantially increased the survival of preterm infants during the last 15 years. There is wider acceptance of the importance of infection as a factor in preterm birth, and increasing recognition that processes leading to preterm birth may be initiated in very early pregnancy (the initiation of pre-eclampsia, major birth defects, premature placental separation), or even prior to pregnancy (prior pregnancy losses). It is unclear whether the familiar clinical presentations of preterm labour and birth reflect different pathophysiological processes. The pathways which link those processes to the consistent pattern of social differences in the probability of preterm birth have prompted new research approaches but in 2002 'the stubborn challenge of preterm birth' remains just that. PMID- 12763105 TI - Neonatal complications following preterm birth. AB - Improvements in neonatal intensive care during the last 20 years have increased the survival of the most immature newborns at 23 weeks from 0% to 65% at some centres, although rates vary widely among neonatal care centres. University of Utah, USA data show that each week in utero after week 23 raises survival by 6 9%, to 90% by 27-28 weeks and 95% by 33 weeks. Provision of care in specialised centres to provide high-risk obstetric and neonatal intensive care, prenatal treatment with corticosteroids, postnatal treatment with surfactant and nitric oxide, and improvements in respirators and equipment to care for extremely immature infants all contribute to these changes. The increased rate of survival for extremely premature newborns has not been accompanied by an increased rate of severe intraventricular haemorrhage or neurological impairment, such as cerebral palsy. Regardless, intraventricular haemorrhage remains a significant problem, especially if associated with post-haemorrhagic hydrocephalus, leading to long term neurological impairment and decreased survival. Necrotising enterocolitis (NEC) is more common in premature than in term newborns and is the most frequent cause of short bowel syndrome in infancy. Survival after surgery for NEC has improved during the last two decades, but complications of nutritional support produce many long-term problems. Retinopathy of prematurity (ROP) remains a frequent cause of neurosensory impairment for extremely premature newborns. Laser photocoagulation for advanced ROP is more effective than cryotherapy for preventing retinal detachment and improving visual outcomes. Despite prenatal corticosteroid treatment and postnatal surfactant administration, many extremely premature newborns still develop bronchopulmonary dysplasia. Abnormal pulmonary function may persist into adulthood, but newer ventilators and management schemes appear to be reducing this long-term morbidity. Many changes in neonatal care occur each year, but carefully controlled outcome studies are needed to evaluate the effectiveness of these newer styles of neonatal intensive care. PMID- 12763106 TI - Economic consequences of preterm birth and low birthweight. AB - The high rates of mortality and morbidity arising from preterm birth and low birthweight impose an immense burden on the health, education and social services and on families. This was evaluated in several economic studies published in the 1970s and early 1980s, but an ability to intervene effectively to increase survival has transformed neonatal care over the past two decades. In addition, the relative prices of resource inputs are vastly different. This paper surveys the scientific literature on the economic consequences of preterm birth and low birthweight, including: (1) studies, which estimate the economic costs of the conditions and (2) economic evaluations of primary and secondary prevention and treatment strategies. This paper summarises the scope and scientific quality of the published evidence, identifies gaps in our knowledge and considers the future research agenda in this area. PMID- 12763107 TI - The impact of multiple preterm births on the family. AB - Multiple births are important contributors to the preterm and low birthweight population and the numbers of twin births have been steadily rising since the early 1980s in all developed countries. This is largely due to the increased use of ovulation induction and multi-embryo transfer in the treatment of subfertility. Parents of preterm twins have been shown to be less responsive to their infants than those with singletons. Parental stress with twins has also been demonstrated by the higher incidence of maternal depression and of child abuse in multiple birth families. Furthermore, siblings of twins are more likely to have behaviour problems. Mortality and long-term morbidity rates are greatly increased amongst multiple birth children. The problems of the single surviving twin and the unaffected co-twin of a disabled child are often underestimated as is the complexity of the bereavement of parents who still have surviving multiples. Addressing the cause of the epidemic of iatrogenic multiple births is likely to be the single most effective way to reduce the number of preterm infants and the long-term problems to which they are prone. PMID- 12763108 TI - Classification and heterogeneity of preterm birth. AB - Three main conditions explain preterm birth: medically indicated (iatrogenic) preterm birth (25%; 18.7-35.2%), preterm premature rupture of membranes (PPROM) (25%; 7.1-51.2%) and spontaneous (idiopathic) preterm birth (50%; 23.2-64.1%). The majority of multiple pregnancies (10% of all preterm births) are delivered preterm (50% for medical reasons). Although medical indications relate more to feto-maternal conditions, PPROM to infections and idiopathic preterm birth to lifestyle, these risk factors are identified in any category, emphasising that preterm birth has a multifactorial origin. Still, several incidences of preterm birth are not completely explained with a plausible cause for PPROM or spontaneous preterm labour suggesting that other causes have yet to be identified. In addition, preterm birth is associated with unrecognised severe congenital anomalies. Variability within the main categories may be explained by the studied population, ethnic group, social class and preventive interventions towards reducing spontaneous preterm birth where the proportion of medically indicated preterm birth is increased. Despite being retrospective a classification according to gestational age at birth is important for neonatal prognosis. Preterm birth is stratified into mild preterm (32-36 weeks), very preterm (28-31 weeks) and extremely preterm (<28 weeks) with increasing neonatal mortality and morbidity. Recent studies suggested that infection was mostly responsible for extreme preterm birth, while stress and lifestyle accounted for mild preterm birth, and a mixture of both conditions contributed to very preterm birth. PMID- 12763109 TI - Gender aspects of preterm birth. AB - It was previously believed that sex differentiation took place when the undifferentiated gonads formed either testes or ovaries. Studies in recent years indicate that sex differentiation begins at conception. The SRY gene on the Y chromosome is already transcribed at the 2-cell stage and triggers growth acceleration in the XY embryos. This accelerated growth is believed to be important for the male embryo as it allows complete testicular differentiation before the levels of oestrogenic hormones become too high as pregnancy progresses. It is well known that the death rate is higher for male than for female fetuses and that the increase is about 30% in chromosomally normal spontaneous abortions (i.e. significantly higher than at birth). National figures from Sweden show that boys are more likely to be delivered prematurely, accounting for 55-60% of all newborns between 23 and 32 gestational weeks. Neonatal deaths in these gestational weeks are also more common among boys. In 1993, the overall 1-year mortality rate (including all gestational weeks) in Sweden was 5.4% for boys and 4.1% for girls. The difference in infant mortality (within 1 year) is most pronounced at extremely early birth (23-24 gestational weeks) being 60% for boys compared with 38% for girls. The release of catecholamines during labour is an important defence mechanism by a hypoxic fetus. Preterm females have significantly higher catecholamine levels than males, which may explain the better outcome in females after a hypoxic event. Deaths occurring secondary to respiratory distress syndrome are greater for males and their cognitive recovery from perinatal intracranial haemorrhage is worse. Pulmonary hypoplasia after preterm rupture of the membranes is significantly more common among male newborns. Gender differences in mode of delivery, fetal heart rate in labour, acidaemia at birth, and age degenerative changes will also be discussed. PMID- 12763110 TI - Mechanisms of labour--biochemical aspects. AB - The mechanism of labour is not fully understood and further research into this important physiological process is needed. In some species, notably sheep, parturition is due to activation of the fetal hypothalamic-pituitary-adrenal axis. However, in primates, this axis appears to have a supportive, rather than essential role. Successful parturition requires an increase in coordinated uterine contractility together with changes in connective tissue that allow cervical ripening and dilatation. In most mammals, however, these changes are synchronised by a fall in maternal progesterone levels and a rise in oestrogens. This is not the case in women in whom the onset of labour occurs without apparent changes in circulating steroid levels. The basis of uterine contractility is the interaction between actin and myosin in myometrial smooth muscle cells. This is driven by calcium through Ca(2+)-calmodulin-dependent myosin light chain kinase (MLCK) activity. Moreover, calcium sensitisation occurs via activation of Rho kinase, a calcium-independent pathway that promotes contractility by inhibiting myosin phosphatase and probably by phosphorylating myosin on the same site as MLCK. Uterine activity can be modulated by many G-protein coupled receptors (GPCRs). For example, receptors coupled to Galpha(q) (oxytocin-, prostanoid FP and TP, endothelin-receptors) stimulate contractility by activating the phospholipase C/Ca(2+) pathway; receptors coupled to Galpha(s) (beta(2) adrenoceptors, prostanoid EP2 and IP, some 5-hydroxytryptamine receptors e.g. 5 HT(7)) relax the uterus by increasing myometrial cyclic AMP levels; and receptors coupled to Galpha(i) (alpha(2)-adrenoceptors, muscarinic, 5-HT(1)) potentiate contractility, probably by inhibiting cAMP production. Because of its relative abundance in pregnant uterine tissue, the oxytocin receptor is an obvious target for tocolytic therapy. Oxytocin antagonists have been introduced into clinical practice for the management of preterm labour and offer the advantage of uterine selectivity and fewer side effects than conventional beta-agonist therapy. PMID- 12763111 TI - The role of oxytocin in parturition. AB - Oxytocin and the oxytocin receptor have two important roles in labour. Evidence in all mammalian species suggests that neurohypophysical oxytocin plays a role in the expulsive phase and, although there are less supporting data, a role for oxytocin in the initiation of labour is likely. The initiation of labour may be mediated in women and rhesus monkeys by paracrine rather than endocrine mechanisms. Although initial characterisation of the oxytocin knockout mouse suggested that oxytocin is not important in this species, subsequent investigations have demonstrated that oxytocin is important for the precise timing of the onset of labour. Studies in knockout mice also confirm important interrelationships between oxytocin and prostaglandins. Oxytocin stimulates prostaglandin release in many species, mainly in the decidua/uterine epithelium. The effects of oxytocin are mediated by tissue-specific oxytocin receptor expression, which leads directly to contraction in the myometrium and prostaglandin formation in the decidua. There is a dramatic increase in oxytocin receptor expression in these tissues in late pregnancy and pharmacological inhibition delays delivery, which suggests that, in contrast to oxytocin, the oxytocin receptor is essential for normal labour. PMID- 12763112 TI - The role of infection in preterm labour. AB - Although there are many maternal characteristics associated with preterm birth, the aetiology in most cases is poorly understood. Our research demonstrates that multiple risk factors, such as maternal age and especially infection, are associated with preterm birth. Bacterial vaginosis and intrauterine infection are now believed to be an important risk factor for preterm delivery. PMID- 12763113 TI - Socio-economic and psychosocial factors in the management and prevention of preterm labour. AB - Socio-economic factors associated with preterm labour include social class, (usually assessed by earnings and education), working conditions (professional status, ergonomic environment, working hours), physical and travelling activities, daily life activities, lifestyle, family status and psychosocial state as related to past and current pregnancy history together with current stress factors. A review of the association of these factors with preterm birth will be reported with an emphasis of the biological plausibility linking mostly emotional, and at a lesser degree, physical and psychological stress to the occurrence of preterm labour. A case control study, carried out in Quebec City among 101 women in preterm labour and 202 matched pregnancies for parity and gestational age, identified 7 risk factors in an explanatory multivariate model among 117 variables: Body mass index (BMI) <20 (OR; 95% CI: 3.96; 2.61-7.09), previous preterm labour (OR; 95% CI: 3.61; 1.12-11.65) previous low birth weight (OR; 95% CI: 2.24; 1.05-7.71), standing at work >2 hours (OR; 95% CI: 3.90; 1.53 9.91), Abruptio placentae (OR; 95% CI: 5.88; 1.20-28.76), urinary tract infection (UTI) (OR; 95% CI: 4.4.3; 1.47-13.34), and stress score >5 (OR; 95% CI: 2.56; 1.20-5.54). The most stressful events were related to family illness, mortality, disruption, violence or financial distress. Some risk factors cannot be modified (previous preterm labour, low birth weight and UTI), while preventive efforts should be directed towards attaining BMI >20 before conception, modifying working conditions during current pregnancy and appropriate management of acute emotional stress. PMID- 12763114 TI - Cervical length assessment by ultrasound as a predictor of preterm labour--is there a role for routine screening? AB - Transvaginal ultrasonography has recently been shown to be an objective, reproducible and reliable method to assess the cervix and predict the risk of preterm delivery in high-risk pregnancies. Assessment of the cervix includes cervical length measurement (CLM) and measurement of dilatation of the internal os in a dynamic functional examination. There is an inverse correlation between cervical length and the frequency of preterm delivery. The high negative predictive value avoids unnecessary interventions such as tocolysis or cerclage in high-risk pregnancies. In contrast, a length of 25 mm or less at 28-30 weeks of gestation is associated with a significantly increased incidence of preterm delivery. Studies in women with high risk for preterm delivery, i.e. contractions, premature rupture of the membranes and history of preterm delivery, have shown a high sensitivity and a high positive predictive value, however in low-risk groups they have failed to show a high sensitivity. From large observational studies in low-risk populations we know that the 50th percentile of the cervical length is 35 mm at 24 weeks of gestation. Advantages of CLM as a screening test include the fact that sonographical assessment of the cervix is a widely accepted and well-standardised method, which requires only a relatively short period of training. Disadvantages of screening are two factors, the first being the low sensitivity of the test and the low prevalence of preterm deliveries in a low-risk population, resulting in cut off values being set at a very low level (i.e. 5th percentile) in order to get acceptable specificity. Secondly, screening is only worthwhile if an effective preventive therapy is available. The debate about tocolysis and cerclage is not yet concluded. Therefore we would not currently recommend cervical length measurement as a screening tool-but as a routine method in high risk gravidas with or without symptoms. Further interest should be focused on scoring systems combining ultrasound with biochemical, endocrinological and maybe molecular cell methods such as the measurement of fetal DNA in maternal blood to prevent preterm deliveries in the general population. PMID- 12763115 TI - Fetal fibronectin--how useful is it in the prediction of preterm birth? AB - OBJECTIVE: To determine the value of cervicovaginal fetal fibronectin as a marker for preterm delivery, a previously published meta-analysis was updated. STUDY DESIGN: Selection criteria confined the analysis to English-language original reports of prospective studies including women at <37 weeks' gestation with intact amniotic membranes. For the outcomes of delivery <37 or <34 weeks' gestation or delivery within 7, 14, or 21 days after fibronectin sampling, we calculated sensitivity and specificity rates for each study, for subgroups of studies, and for all studies combined. RESULTS: A total of 40 studies were included. Statistical heterogeneity was seen in the majority of calculations of combined results and a random-effects model was used in these cases. For the outcomes of delivery <37 and <34 weeks' gestation, overall sensitivity rates were 52% and 53%, and overall specificity rates were 85% and 89%, respectively. For the outcomes of delivery within 7, 14, and 21 days, we calculated sensitivity rates of 71%, 67%, and 59% and specificity rates of 89%, 89%, and 92%, respectively. For the subgroup of women with symptoms of preterm labour, sensitivity rates for delivery <37 and <34 weeks' gestation or delivery within 7, 14, and 21 days of 54%, 63%, 77%, 74%, and 70% and specificity rates of 85%, 86%, 87%, 87%, and 90% were calculated. CONCLUSIONS: Cervicovaginal fetal fibronectin is an effective short-term marker of preterm delivery, especially in women with symptoms of preterm labour. Because results appear to be heterogeneous in different studies, caution should be taken when they are applied to a specific population. PMID- 12763116 TI - Infection in the prediction and antibiotics in the prevention of spontaneous preterm labour and preterm birth. AB - The association between infection and spontaneous preterm labour is now well established and thought to be responsible for preterm birth in up to 40% of cases. Preterm labour that is due to infection is refractory to the use of tocolytic agents. So the knowledge that infection may be the cause is unhelpful once a woman is admitted in spontaneous preterm labour, since by that time there may be irreversible changes in the uterine cervix, which renders futile those attempts to inhibit the process. It would be much more logical to use the association between infection and spontaneous preterm labour to identify a group of women at risk and to intervene using antibiotic prophylaxis. It is important to record, that the earlier in gestation at which abnormal genital tract colonisation is detected, the greater is the risk of an adverse outcome. For example, abnormal genital tract flora at 26-32 weeks gestation is associated with preterm birth with an odds ratio (OR) of 1.4 to 2, whereas abnormal genital tract flora at 7-16 weeks gestation carries an OR of 5 to 7.5. Intervention studies have used different antibiotics in different dosage regimes by different routes of administration to patients of differing risks at different gestational ages. Not surprisingly this has led to differing results. If intervention is to be successful, the antibiotics chosen should be active against bacterial vaginosis or bacterial vaginosis-related organisms and should be used early in pregnancy in those women with the greatest degree of abnormal genital tract flora. While there is logic in using intravaginal antibiotics to deliver a heavy antibiotic load to the vagina where heavy abnormal colonisation exists, there is also logic in considering systemic antibiotics to eradicate those organisms, which have already gained access to the decidua. It may be that the greatest chance of benefit would exist if both routes of administration were combined. Yet no study has evaluated the combination of both intravaginal and systemic antibiotics to eradicate abnormal genital tract flora for the prevention of preterm birth. PMID- 12763117 TI - Antenatal corticosteroids--current thinking. AB - Numerous investigations have indicated that antenatal corticosteroid administration reduces neonatal morbidity in preterm infants. Despite this, antenatal corticosteroid usage continued to be low on both sides of the Atlantic up to the late 1980s and early 1990s. Antenatal corticosteroid therapy finally came of age in 1995 with the publication of the National Institutes of Health Consensus Statement on effects of corticosteroids for fetal maturation on perinatal outcomes. The randomised trials of antenatal corticosteroid therapy have been based on the use of dexamethasone or betamethasone. However, observational evidence indicates differences in outcome, which may relate to the preparations themselves or to the preservatives included. PMID- 12763118 TI - Problems and challenges in the management of preterm labour. AB - The main problem with preterm labour is our lack of progress in the successful management of this condition. We need to reassess our approach to this problem because preterm labour is not a disease, but an event, which may result from multiple independent pathways. This problem has also been affected significantly by medical advances such as infertility treatments and changes in neonatal survival at the limit of viability. The specific challenges that we face in managing preterm labour include: problems with definition; aetiology, including genetic and infection components; diagnostic problems, such as true versus false labour and role of cervical length and fetal fibronectin; and specific interventions according to the antepartum, intrapartum and postpartum challenges. In order to address the main issue, and make future progress in the management of preterm labour, we should consider the implementation of a 'Postpartum Preterm Labour Diagnostic Workup Protocol'. These data/workup results could be entered on web-based databases for each preterm labour 'event'. An international research team could analyse data relating to specific aetiological patterns and subgroup analyses, leading to the collaborative development of 'aetiology specific' management modalities. This approach requires a close collaboration between clinicians and researchers, in order to make significant progress in this difficult area, and ultimately improve perinatal outcomes. PMID- 12763119 TI - Recognition of preterm labour as a set of 'complex diseases' increases efficacy of tocolytic treatment. PMID- 12763120 TI - Method of delivery for the preterm infant. AB - The principal issue concerning the mode of delivery is that while elective caesarean births may reduce the chances of fetal or neonatal death, this method of delivery might also increase maternal morbidity. Six trials with a total of 122 women investigated the effects of a policy of elective caesarean delivery (ECD) versus expectant management for small babies. A meta-analysis of these six trials also included additional unpublished data from several of the authors. Babies in the 'elective' group were less likely to develop respiratory distress syndrome and were more likely to have a low pH after birth. They were also less likely to have neonatal seizures and there were fewer perinatal deaths, although the studies were too small to detect even very large differences. However, there was significantly more serious maternal morbidity. There is insufficient evidence to evaluate a policy of ECD. All trials described major problems in recruitment and no subsequent trials have been identified. Changes in population-based data suggest that ECD of very preterm infants has increased markedly in the last 20 years. Given these changes, it seems unlikely that recruitment for future trials will be any easier than it was in the recent past. PMID- 12763121 TI - The history of tocolysis. AB - In 1950, the World Health Organisation (WHO) defined prematurity as a birthweight of 2500 g or less and in 1961 as a gestational age of less than 37 weeks. The time in between marks an era in which there was growing recognition of the importance of gestational age at birth and how to influence it. The latter was facilitated too by the development of tocography, which permitted some semi objective measurement of uterine contractility. Along with it, came a growing interest in agents that could control uterine contractility beyond the earlier classical approaches of hormones and gastrointestinal spasmolytics. Hence, the early 1960s saw much research interest in agents, such as nylidrine, isoxsuprine, and orciprenaline that could suppress uterine contractility as one of their many beta-agonist properties. Subsequently, two approaches would be used to shift the balance towards uterine function over and above the influence on other bodily functions. One consisted of supplementing these drugs with agents, such as calcium antagonists and beta-receptor blockers that were hoped to suppress non uterine actions. The other was a search for drugs in the same class with greater uterospecificity and more selective binding to uterine as opposed to other receptors. Neither of these approaches has ever fully fulfilled the hopes that were pinned on them, but they resulted in the availability of a large number of agents to suppress uterine contractility. The advent of prostaglandins as regulators of uterine contractility and the ability to suppress their biosynthesis saw another range of attempts to suppress uterine activity. They included aspirin, sodium salicylate, flufenamic acid, sulindac and indomethacin, but some were clearly based on a defective understanding of how uterine prostaglandin synthesis can be influenced. In the meantime, a flurry of other agents came and went, often more than once, testifying to the ingenuity of clinicians in trying to solve a problem that is poorly understood. Some, such as relaxin and ethanol, came and disappeared. Others, such as calcium antagonists, entered the scene as protectors against the non-uterine effects of other agents, went, and re-entered the scene in their own right. Still others, such as magnesium sulphate, came, lingered around, and became credited with effects in preterm labour that do not depend on affecting uterine contractility. Amidst this all arose the term tocolysis, coined in 1964 by Mosler from the Greek stems 'tauomicronkappaomicronzeta' and 'lambdaupsilonepsiloniotanu', to epitomise all of this ingenuity. PMID- 12763122 TI - The case for tocolysis in threatened preterm labour. AB - The failure of tocolytics to improve neonatal outcomes in placebo-controlled trials has wrongly been interpreted as evidence that they do not work. While delivery is unequivocally prolonged by 24 hours, 48 hours and 7 days, the time gained was not exploited to optimise neonatal outcome. These trials typically studied women at relatively advanced gestational ages with predictably good outcomes, enrolled them in tertiary centres where they could not benefit from in utero transfer, and had low levels of corticosteroid administration. No study has been powered to detect clinically meaningful differences that might be expected to accrue from 1-7 days prolongation of gestation. Despite this, Bayesian interpretation suggests that tocolytics do improve neonatal outcome. The largest placebo-controlled study showed clear trends towards better survival in fetuses <28 weeks, lower rates of cerebral palsy and higher Bayley mental scores. Meta analysis of neonatal morbidity in the beta-agonist trials suggests a near significant reduction in respiratory distress syndrome (RDS), together with trends towards reduced intraventricular haemorrhage, necrotising enterocolitis, and patent ductus arteriosus. Finally, there is the Orwellian analogy that tocolytics don't work, but some work better than others. Although calcium antagonists have not been evaluated against placebo, meta-analysis of comparative trials with beta-agonists demonstrate a significantly lower incidence of RDS and neonatal jaundice, presumably mediated through the reduced chance of delivery within 48 hours and 7 days. Development of tocolytics that are safe for mother and baby should facilitate adequately-powered placebo-controlled studies, which both focus on women most likely to benefit and capitalise on the 1-7 days gained. PMID- 12763124 TI - Tocolysis in the management of multiple pregnancy. PMID- 12763123 TI - What are the realistic expectations of tocolytics? AB - Preterm birth, the leading cause of neonatal morbidity and mortality worldwide, is a major public health problem in terms of loss of life, long term disability (e.g. cerebral palsy, chronic lung disease), and health-care costs. The outcome of preterm infants is directly related to the gestational age at delivery. The goal of tocolytic therapy is to reduce neonatal morbidity and mortality by delaying delivery, to allow for the administration of corticosteroids and/or the safe transfer to a tertiary-care centre. However, currently available tocolytics in Canada (ritodrine, indomethacin, calcium antagonists, magnesium sulphate) have poor efficacy, have not been shown to increase the completion of a course of corticosteroids, are potentially associated with significant maternal/fetal side effects, and most importantly, have not been shown to improve neonatal outcomes. At the Canadian Tocolysis Consensus Conference, there was general agreement that recommendations should be based on good quality research evidence, particularly that of randomised clinical trials when available. It was concluded that there is little evidence to support the use of any of the currently available tocolytics; tocolytic use has not been associated with improved perinatal outcomes and often have detrimental effects on the mother. Therefore, questionable efficacy and potentially serious side effects may outweigh their use. Any new tocolytic demonstrated to improve neonatal outcome will have an immediate impact on societal and long term public health-care costs. PMID- 12763125 TI - The development and introduction of anti-oxytocic tocolytics. AB - The perfect tocolytic agent, which is completely safe for both the mother and fetus and, which will inhibit uterine contractions and stop preterm labour in every case does not exist and the search continues. Recently, research into a new group of tocolytic agents (the oxytocic antagonists) has led to the introduction of a new licensed drug, atosiban. Since the early 1950s, modifications of the oxytocin molecule have resulted in many analogues and antagonists, though initially none emerged as potentially useful drugs. Further modifications resulted in full uterotonic antagonism in animal models before an analogue was found that inhibited vasopressin-stimulated uterine contractions in non-pregnant healthy women. In vitro and animal models suggested the molecule was fully antagonistic, although it was found to be only partially agonistic in women. Further developments led to two modified oxytocin molecules with higher receptor affinity for human myometrium, both of which lacked agonism in humans. The analogue, atosiban, was found to be more potent and so was chosen for clinical evaluation in dysmenorrhoea and preterm labour. The first clinical reports were open label, observational pilot studies. Randomised, double-blind, phase II placebo-controlled studies followed showing that atosiban was significantly more effective than placebo with very few side effects. Dose-response studies and phase III studies in which study or placebo groups could use alternative tocolytic agents also suggested that atosiban was an effective tocolytic agent with very few adverse events. The recent worldwide comparative study of atosiban versus different beta-agonists represents the largest and most strictly controlled study of tocolytics ever published. Atosiban was found to be at least as effective as the beta-agonists as a tocolytic agent, but significantly less likely to result in maternal cardiovascular side effects or the need to discontinue therapy as a result of unacceptable side effects. PMID- 12763126 TI - Exploring the role of Tractocile in everyday clinical practice. AB - The purpose of this retrospective study is to evaluate the effects of atosiban (Tractocile available in Austria since February 2000) for routine treatment of women with threatened preterm delivery. The advantage of this drug compared to other tocolytic agents is its specific action on reproductive tissues without the accompanying severe side effects. Women (n = 208) were retrospectively evaluated. Diagnoses at admission were preterm labour (n = 117), preterm rupture of membranes (n = 65), incompetent cervix (n = 19) and vaginal bleeding (n = 7). Gestational age was between weeks 21 and 33 of pregnancy. Preterm labour was defined as >/=4 uterine contractions/30 min and cervical length <30 mm examined by vaginal ultrasound and/or detection of vaginal fetal fibronectin. Tocolytic effectiveness was determined as the number of women having a diagnosis of preterm labour who were still pregnant after 48 hours and after 7 days. The influence on the frequency of contractions before and 3-12 hours after the start of treatment was assessed. Maternal side effects, perinatal and neonatal morbidity and transfers to the NICU were also evaluated. The proportion of women who remained undelivered was 78.7% after 48 hours, and 64.3% after 7 days. Atosiban decreased the frequency of contractions from 5.4/30 min before treatment to 1.6 contractions/30 min after the start of treatment. At the initial bolus application, 20.2% of women presented drug-related side effects, such as nausea, vertigo and flush over a short period of 1-2 minutes. During infusion, side effects possibly related to atosiban could be detected in 6% of women. Mean length of stay was 11.8 days in the NICU and 30.9 days in intermediate care. Twenty-three children developed intraventricular haemorrhage (I-IV). In conclusion, atosiban is an effective tocolytic drug in the treatment of preterm labour and preterm rupture of the membranes. It has significantly less side effects due to its lack of cardiovascular activity. PMID- 12763127 TI - Genetic factors in preterm birth. PMID- 12763128 TI - Role of cytokines and other inflammatory mediators. AB - In spite of impressive advances in biochemistry and molecular biology, it has not yet been possible to fit the individual biochemical components of cervical ripening and dilatation to a uniform clinical moiety or to uncover any regulatory mechanisms. The production of interleukin-8 by activated fibroblasts and macrophages plays a key role in cervical ripening, since this cytokine induces chemotaxis, activation, and degranulation of neutrophilic granulocytes with the consequent release of various proteases, including collagenase. In addition, the extravasation of neutrophilic granulocytes is mediated-as in the early stage of an acute inflammatory reaction-by a brief increase in adhesiveness of vascular endothelium. This is known to be modulated by the cytokine-induced increase in the expression of endothelial adhesion molecules. Furthermore, an increase in pro inflammatory cytokine and proteinase concentrations in preterm delivery seems to occur at earlier stages of cervical dilatation than in term delivery. It is also well known that in patients with chorioamnionitis, the levels of pro-inflammatory cytokines are elevated in amniotic fluid, maternal serum, cervical secretion, placenta, and other compartments of the placento-maternal unit, and are associated with preterm uterine contractions. We have demonstrated for the first time that cytokine concentrations in the lower uterine segment in patients with chorioamnionitis are strongly elevated. We conclude from our data that increased concentrations of pro-inflammatory cytokines may also play a pivotal role in cervical softening and dilatation during chorioamniotic infection. Our data agree with the hypothesis of Liggins who stated nearly 20 years ago that cervical ripening may be an inflammatory reaction, which leads to increased prostaglandin synthesis, preterm labour and finally to preterm delivery. PMID- 12763129 TI - Intrauterine infection and the development of cerebral palsy. AB - Cerebral palsy is a serious motor disorder that appears in early life. The expectation that improved obstetrical and neonatal care would decrease the rate of this condition has not been realised. Recent evidence indicates that white matter brain lesions, often termed periventricular leukomalacia (PVL), are the most important identifiable risk factors for the development of cerebral palsy. The hypothesis under examination is that inflammatory cytokines released during the course of intrauterine infection play a central role in the genesis of preterm parturition, fetal PVL, and cerebral palsy. We examined the relationship between umbilical cord plasma concentrations of cytokines at birth and the occurrence of PVL in preterm gestation and demonstrated that umbilical cord plasma concentrations of interleukin (IL)-6 was a significant independent predictor of PVL-associated lesions. We also demonstrated that preterm neonates born to mothers with elevated amniotic fluid concentrations of pro-inflammatory cytokines were at increased risk for the subsequent development of PVL and cerebral palsy. Histological chorioamnionitis and congenital neonatal infection related morbidity were more common in neonates with PVL than those without PVL in this study. We have also been able to induce PVL-like brain white matter lesions in the fetal rabbit after experimental ascending intrauterine infection. In support of this hypothesis, we were able to demonstrate overexpression of tumour necrosis factor-alpha and IL-6 in histological sections of neonatal brains with PVL. Moreover, the presence of funisitis, a histological counterpart of the fetal inflammatory response syndrome, and elevated concentrations of amniotic fluid IL 6 and IL-8 were strong and independent risk factors for the subsequent development of cerebral palsy at the age of 3 years in our recent study. Therefore, clinical and experimental data provide strong support for the hypothesis. There are significant implications of our findings. First, cytokine determinations in amniotic fluid provide information about the risk of PVL and cerebral palsy before birth. Second, the process responsible for some cases of PVL and cerebral palsy begins during intrauterine life, implying that effective strategies for the prevention of cerebral palsy associated with PVL must begin in utero. PMID- 12763130 TI - The critical role of perinatal pathology. AB - In spite of great advances in imaging and biochemistry, histological examination of tissues remains a vital part of the multidisciplinary approach to the prevention of the onset, morbidity and mortality of preterm birth. There has been increasing interest in the role of infection and inflammatory cytokines in causation both of early labour and the white matter damage in the brain of preterm infants. However, labour itself is associated with the build up of increased numbers of inflammatory cells in the uterine cervix and increased concentrations of inflammatory cytokines and positive microbiological cultures may reflect carriage or contamination. Confirmation of an infective aetiology in an individual case is best achieved by demonstration of a pathological inflammatory response in tissues, for example, by showing the presence of chorioamnionitis in the placenta. A proper understanding of the poor response to neonatal intensive care of some preterm babies often requires histological examination of the lungs after death, where unsuspected pneumonia, interstitial emphysema and/or pulmonary hypoplasia may help provide an explanation for the adverse outcome in individual cases. The pathophysiological mechanism of brain injury in preterm infants is undergoing re-evaluation, and the systemic study of brain tissue using the latest histological techniques may elucidate the importance of apoptosis in this situation and could point the way towards an effective preventative strategy. Paediatric pathology is also essential to explain many cases of sudden unexpected death in preterm infants, as demonstrated by the recent realisation that death may be caused by total parenteral nutrition fluid-associated myocardial necrosis, and acute cardiac tamponade. PMID- 12763131 TI - Looking to the future. AB - Since the 7th and 13th Study Groups of the Royal College of Obstetricians and Gynaecologists met in 1977 and 1985, respectively, no meeting of this magnitude has convened to discuss the problems of spontaneous preterm labour and delivery and the associated fetomaternal mortality and morbidity. In the 17 years or so since that time, advances have been made in our understanding of the mechanisms of labour, the role of infection, the benefit of antepartum corticosteroids and the development of safer more specific tocolytics. In the future, an understanding of the genetic risk of spontaneous preterm labour and preterm birth is essential, particularly with respect to the predisposition to produce potentially damaging pro-inflammatory cytokines. The examination of the tissue damage will require pathologists specifically trained in perinatal pathology if the aetiology is to be ascertained and future management tailored to the risks. A greater understanding of fetomaternal immunology and response to antigen exposure in pregnancy may help us to understand which fetomaternal pairs are at greatest risk of responding by delivering preterm, with greater or lesser tissue damage than others with similar risk. Specifically, the relation between spontaneous preterm labour and proteinuric pre-eclampsia with their common immunology, inflammatory response and tissue damage leading to either spontaneous preterm labour or iatrogenic preterm birth will need to be addressed. This meeting has been very clinically and obstetrically orientated, in future we will need to involve epidemiologists, neonatologists, microbiologists, genito-urinary medicine physicians, immunologists, geneticists, biochemists, physiologists and endocrinologists. Although spontaneous preterm labour and preterm birth are the major causes of perinatal mortality and morbidity in the developed world, the definition and management protocols for spontaneous preterm labour varies from unit to unit and country to country. A process has already begun, hopefully fuelled by this meeting and those attending, to develop an international consensus on definitions and evidence-based practical guidelines on the management of spontaneous preterm labour. Perhaps in the longer term it may be possible to influence standards of care, outcome measures and training across international boundaries. PMID- 12763132 TI - Extrathymic T-lymphocyte development. PMID- 12763133 TI - p16(INK4a) induces differentiation and apoptosis in erythroid lineage cells. AB - OBJECTIVE: Hematopoiesis is regulated by proliferation, differentiation, and death. p16(INK4a) has been reported to regulate apoptosis and differentiation of diverse cells, as well as arresting the cell cycle at G1 phase. The aim of this study is to explore the properties of p16 in apoptosis and differentiation of erythroid cells. METHODS: We transfected the INK4a gene to K562 cells, which defect the INK4a gene, and compared the effect of enforced expression of p16(INK4a) with that of various additives, topoisomerase I inhibitor (SN 38), interferon-alpha, phosphatidyl-inositol-3 kinase inhibitor (LY294002), and serum deprivation, which arrest cell cycle at different phases. We also investigated the role of p16(INK4a) in normal day-6 human erythroid colony-forming cells by transfecting the INK4a gene. RESULTS: p16(INK4a) induced cell cycle arrest at the G0/G1 phase, and promoted erythroid differentiation in viable K562 cells, but induced apoptosis in K562 cells with incomplete differentiation. The apoptosis induced by p16 was accompanied with downregulation of bcl-x and nuclear NF kappaB. These findings were not observed in K562 cells treated with various additives. p16(INK4a) decreased the cell viability and promoted apoptosis in day 9 ECFC. CONCLUSION: We propose that p16(INK4a) plays a role in maintaining homeostasis during erythroid differentiation, and that the mechanisms for this effect are not confined to those inducing cell cycle arrest. PMID- 12763134 TI - The inhibitory effect of anti-CD33 monoclonal antibodies on AML cell growth correlates with Syk and/or ZAP-70 expression. AB - OBJECTIVES: Acute myeloid leukemia (AML) cells express the cell surface antigen CD33 that can function as a downregulator of cell growth, mediating growth arrest and apoptosis. The protein kinase Syk is an essential element in several cascades coupling certain antigen receptors to cell responses. Recently we reported that CD33 recruits Syk for its signaling in AML cell lines. In this study, we further investigated the mechanism(s) of Syk engagement in CD33 signaling in primary AML samples. METHODS: We investigated 25 primary AML samples for their proliferative response (3H-thymidine incorporation) and biochemical changes (Western blot analysis) to anti-CD33 mAb treatment. RESULTS: Proliferation studies demonstrated that 14 (56%) of AML samples were responsive (R) while 11 (44%) were nonresponsive (n-R) to inhibitory antibody activity. Seven of 25 AML samples (28%) expressed undetectable levels of Syk. However, cells from two of these patients expressed the ZAP-70 protein kinase. In Syk/ZAP-70(+) samples, CD33 ligation inhibited proliferation in 70% of cases, while none of the Syk/ZAP-70(-) samples was responsive. There were significant biochemical differences between responder and nonresponder AML populations. In responder samples, CD33 ligation induced phosphorylation of CD33 andSyk and formation of the CD33/Syk complex. In nonresponder samples, CD33 was not phosphorylated, and Syk was in complex with the SHP-1 protein phosphatase constitutively. CONCLUSIONS: Syk is an important component in the regulation of proliferation in AML cells. The differential response of AML cells to CD33 ligation is associated with the level of the Syk expression. PMID- 12763135 TI - Cellular and molecular abnormalities in severe congenital neutropenia predisposing to leukemia. AB - Severe congenital neutropenia (SCN) is a rare hematological disease characterized by a selective decrease in the level of circulating neutrophils in peripheral blood, maturation arrest at the promyelocyte stage of differentiation in the bone marrow, recurrent severe infections, and evolution to acute myelogenous leukemia (AML). Cellular and molecular studies of 12 SCN patients, including 5 patients that evolved to develop AML, revealed impaired proliferative characteristics and accelerated apoptosis of bone marrow progenitor cells in SCN compared with 11 healthy controls as demonstrated by flow cytometry analysis. Sequencing analysis revealed heterozygous deletion or substitution mutations in the neutrophil elastase (NE) gene in 9 of 12 patients but not in healthy controls. Expression of various NE mutants, but not normal NE, resulted in accelerated apoptosis of human promyelocytic HL-60 progenitor cells, similar to impaired survival observed in patients' cells. Bone marrow-derived primitive CD34(+) and CD33(+)/CD34(-) progenitor cells from SCN patients evolving to AML, all with mutations in the granulocyte colony-stimulating factor receptor (G-CSFR) gene, demonstrated normal cell survival, whereas more differentiated CD15(+)/CD33(-)/CD34(-) cells negative for mutant G-CSFR gene, continue to exhibit accelerated apoptosis. These data demonstrate that impaired survival of bone marrow myeloid progenitor cells, probably driven by expression of mutant NE, is the cellular mechanism responsible for neutropenia in SCN. Furthermore, our results suggest that acquired G-CSFR mutations may initiate signaling events that override the pro-apoptotic effect of mutant NE in primitive progenitor cells, resulting in an expansion of the abnormal AML clone. PMID- 12763136 TI - Characterization of Siglec-5 (CD170) expression and functional activity of anti Siglec-5 antibodies on human phagocytes. AB - OBJECTIVE: The Siglec family of proteins consists of at least 10 members with immunoglobulin and lectin domains and with similar sialic acid-binding properties. Many Siglec family members are expressed on hematopoietic cells and are involved in cell/cell interactions. Some family members are suspected of regulating cellular processes through specific signaling pathways. Monoclonal antibodies were generated against specific epitopes of Siglec-5 (CD170) and were used to determine expression of Siglec-5 on normal blood and marrow cells and cell lines. The antibodies also were used to elucidate functional activity for Siglec-5 on blood neutrophils. METHODS: Flow cytometry and ELISA were used to determine the specificity of the monoclonal antibodies for Siglec-5 and to determine expression patterns. Chemiluminescence assays were employed to measure the oxidative burst activity of whole blood or purified neutrophils following treatment with the anti-Siglec-5 antibodies. RESULTS: Cell surface expression analysis demonstrated that the protein was expressed on gated human neutrophil and monocyte populations, both in the blood and bone marrow. Expression on neutrophils was enhanced by one-hour treatment with fMLP or TNF-alpha. Epitope specific anti-Siglec-5 monoclonal antibodies did not directly activate human neutrophils; however, antibody binding augmented neutrophil oxidative burst activity as determined by fMLP-induced luminol-dependent chemiluminescence. CONCLUSION: Data demonstrating expression of Siglec-5 on cells of the myelomonocytic lineage and alteration of its expression by inflammatory stimuli suggest a role for this protein in cell/cell interactions following microbial exposure. PMID- 12763137 TI - Stathmin expression and megakaryocyte differentiation: a potential role in polyploidy. AB - OBJECTIVE: Megakaryopoiesis is characterized by two major processes, acquisition of lineage-specific markers and polyploidization. Polyploidy is a result of endomitosis, a process that is characterized by continued DNA replication in the presence of abortive mitosis. Stathmin is a major microtubule-regulatory protein that plays an important role in the regulation of the mitotic spindle. Our previous studies had shown that inhibition of stathmin expression in human leukemia cells results in the assembly of atypical mitotic spindles and abnormal exit from mitosis. We hypothesized that the absence of stathmin expression in megakaryocytes might be important for their abortive mitosis. MATERIALS AND METHODS: The experimental models that we used were human K562 and HEL cell lines that can be induced to undergo megakaryocytic differentiation and primary murine megakaryocytes generated by in vitro culture of bone marrow cells. The megakaryocytic phenotype was evaluated by flow cytometry and light microscopy. The DNA content (ploidy) was analyzed by flow cytometry. Stathmin expression was analyzed by Western and Northern blotting and by RT-PCR. RESULTS: Our studies showed an inverse correlation between the level of ploidy and the level of stathmin expression in megakaryocytic cell lines and in primary cells. More importantly, inhibition of stathmin expression in K562 cells enhanced the propensity of these cells to undergo endomitosis and to become polyploid upon induction of megakaryocytic differentiation. In contrast, inhibition of stathmin expression interfered with the ability of the cells to acquire megakaryocyte specific markers of differentiation. CONCLUSION: Based on these observations, we propose a model of megakaryopoiesis in which stathmin expression is necessary for the proliferation and differentiation of early megakaryoblasts and its suppression in the later stages of megakaryocytic maturation is necessary for polyploidization. PMID- 12763138 TI - Erythropoietin-induced serine 727 phosphorylation of STAT3 in erythroid cells is mediated by a MEK-, ERK-, and MSK1-dependent pathway. AB - OBJECTIVE: Erythropoietin (EPO) is a key regulator of erythropoiesis, playing a role in both the proliferation and differentiation of erythroid cells. One of the signal transduction molecules activated upon EPO stimulation is signal transducer and activator of transcription (STAT) 3. Besides tyrosine 705 phosphorylation of STAT3, serine 727 phosphorylation has been described upon EPO stimulation. In the present study, we investigated which molecular pathways mediate the STAT3 serine 727 phosphorylation and the functional implications of this phosphorylation. METHODS: The EPO-dependent erythroid cell line ASE2 was used to investigate which signaling routes were involved in the STAT3 serine 727 phosphorylation. Western blotting using phosphospecific antibodies was used to assess the phosphorylation status of STAT3 molecules. Transfection analysis was performed to investigate the transactivational potential of STAT3, and quantitative RT-PCR was used to study the in vivo gene expression of STAT3-responsive genes. RESULTS: Western blotting of extracts of cells exposed to various chemical inhibitors revealed that the MEK inhibitors PD98059 and U0126 abrogated the EPO-mediated STAT3 serine 727 phosphorylation without an effect on tyrosine phosphorylation. Further analysis showed that MSK1 is activated downstream of ERK, and retroviral transductions with kinase-inactive MSK1 revealed that MSK1 is necessary for STAT3 serine phosphorylation. Furthermore, the STAT3-mediated transactivation was reduced by blocking the STAT3 serine phosphorylation with the MEK inhibitor U0126 or by expression of kinase-inactive MSK1. CONCLUSIONS: The EPO-induced STAT3 serine 727 phosphorylation is mediated by a pathway involving MEK, ERK, and MSK1. Furthermore, serine phosphorylation of STAT3 augments the transactivational potential of STAT3. PMID- 12763139 TI - Reversible expression of CD34 by adult human bone marrow long-term engrafting hematopoietic stem cells. AB - OBJECTIVE: We previously reported that CD34(-) population of bone marrow (BM) cells from adult humans contains cells capable of engraftment and multilineage differentiation. We also reported on the reversibility of CD34 expression by murine hematopoietic stem cells. Based on long-term observations in primary, secondary, and tertiary sheep recipients, we now present definitive evidence for the long-term engrafting capability of human BM CD34(-) cells, and the reversibility of CD34 expression by human BM hematopoietic stem cells (HSC) in vivo. MATERIALS AND METHODS: We used serial transplantations into primary, secondary, and tertiary preimmune fetal sheep recipients to evaluate and compare the long-term engraftment and differentiation of adult human bone marrow-derived CD34(-) and CD34(+) cells in vivo. RESULTS: In primary hosts CD34(-) or CD34(+) cells produced multilineage human cell activity that persisted for 31 months. To confirm the long-term engrafting characteristics of CD34(-) cells and determine whether CD34 expression on human HSC is reversible, we transplanted human CD34(-) and CD34(+) cells obtained from primary hosts into secondary sheep recipients. Multilineage engraftment occurred in all secondary hosts, and in tertiary hosts transplanted with CD34(-) or CD34(+) cells obtained from BM of secondary recipients. CONCLUSION: These results demonstrate that human BM CD34(-) cells are capable of long-term multilineage engraftment in vivo. The finding that both CD34(-) and CD34(+) cells from primary/secondary groups engraft secondary/tertiary hosts indicates that CD34 expression on human HSC is reversible, a process that does not impair HSC function in vivo. PMID- 12763140 TI - Cotransplantation of human mesenchymal stem cells enhances human myelopoiesis and megakaryocytopoiesis in NOD/SCID mice. AB - OBJECTIVE: For approximately 5% of autologous transplant recipients and a higher proportion of allogeneic transplant recipients, low level and delayed platelet engraftment is an ongoing problem. Mesenchymal stem cells (MSC), which can be derived from bone marrow as well as other organs, are capable of differentiation into multiple cell types and also support hematopoiesis in vitro. Because cotransplantation of marrow-derived stromal cells has been shown to enhance engraftment of human hematopoietic stem cells, we hypothesized that cotransplantation of MSC could enhance platelet and myeloid cell development. MATERIALS AND METHODS: We tested this hypothesis by transplantation of CD34 selected mobilized human peripheral blood stem cells (PBSC) into sublethally irradiated NOD/SCID mice with or without culture-expanded human MSC and evaluated human myeloid, lymphoid, and megakaryocytic engraftment with flow cytometry and in vitro cultures. RESULTS: We find that MSC cotransplantation enhances human cell engraftment when a limiting dose (<1 x 10(6)) of CD34 cells is administered. This enhancement is characterized by a shift in the differentiation of human cells from predominantly B lymphocytes to predominantly CD13(+), CD14(+), and CD33(+) myeloid cells with a corresponding increase in myeloid CFU in the marrow. Megakaryocytopoiesis is enhanced by MSC cotransplantation as assessed by an increase in both marrow CFU-MK and circulating human platelets. In contrast, MSC do not affect the percentage of human bone marrow cells that expresses CD34(+). CONCLUSIONS: Cotransplantation of human mesenchymal stem cells with CD34(+) selected hematopoietic stem cells enhances myelopoiesis and megakaryocytopoiesis. PMID- 12763141 TI - Differences in heat sensitivity between normal and acute myeloid leukemic stem cells: feasibility of hyperthermic purging of leukemic cells from autologous stem cell grafts. AB - OBJECTIVES: In autologous stem cell transplantation contamination of the graft with malignant cells is frequently noticed and necessitates the use of in vivo or in vitro purging modalities. The hematopoietic recovery after transplantation depends on the number of stem and progenitor cells in the transplant. Therefore, in the present study the effects of hyperthermic treatment on the human normal and acute myeloid leukemic (AML) stem cell compartment were investigated. METHODS: Normal bone marrow and AML blasts were heat treated up to 120 minutes at 43 degrees C. The surviving fractions of the different stem cell subsets were determined using in vitro methylcellulose and cobblestone area-forming cell (CAFC) clonogenic assays, as well as the in vivo NOD/SCID repopulating assay. The leukemic nature of the colonies from AML cells was confirmed by RT-PCR analysis. In order to increase the therapeutic index of the hyperthermic purging modality, the heat treatment was preceded by a 3-hour incubation at 37 degrees C with the ether lipid ET-18-OCH(3) (25 microg/mL). RESULTS: It could be demonstrated that normal progenitor cells are far more resistant to hyperthermia than leukemic progenitor cells (56%+/-7% vs 9.9%+/-2.6% survival after 60 minutes at 43 degrees C, respectively). Furthermore, normal hematopoietic stem cells appear to be extremely resistant to the heat treatment (94%+/-9% survival after 60 minutes at 43 degrees C). In contrast, in the leukemic stem cell compartment no significant differences in heat sensitivity between the stem cells and progenitor subsets could be observed (12.3%+/-2.9% vs 9.9%+/-2.6% survival after 60 minutes at 43 degrees C, respectively). The combined treatment resulted in a survival for normal progenitor and stem cells of 32%+/-6% and 85%+/-15% after 60 minutes at 43 degrees C, respectively. Under these conditions the number of leukemic stem cells was reduced to 1%+/-0.3%. After 120 minutes at 43 degrees C, no AML-colonies could be detected anymore. CONCLUSIONS: Our data demonstrate that leukemic stem cells have an increased hyperthermic sensitivity compared to their normal counterparts and that this difference can be further increased in combination with ET-18-OCH(3). These striking differences in heat sensitivity warrant the use of hyperthermia as a clinically applicable purging modality in autologous stem cell transplantation. PMID- 12763143 TI - [Immunohematologic follow-up and transfusion safety in hematopoietic stem cells allografts]. AB - Since 1990, specific documentation is established between the immunohematology laboratory of the French EFS Lorraine-Champagne and the marrow transplantation unit of the CHU of Nancy, for patients undergoing hematopoietic stem cells transplantation. These documents and the standardization of the immunohematologic follow-up of those patients, have contributed to the improvement of the transfusion safety. PMID- 12763142 TI - Intravenous busulfan-based conditioning prior to allogeneic hematopoietic stem cell transplantation: myeloablation with reduced toxicity. AB - OBJECTIVE: Allogeneic transplantation is a potentially curative treatment for hematologic malignancies but is associated with a high rate of complications. Busulfan is a common component of pretransplant conditioning but has an erratic and unpredictable bioavailability when administered orally. Intravenous (IV) busulfan was recently introduced into clinical practice. Prior studies showed consistent and predictable drug delivery with tight control of busulfan plasma levels, avoiding over- and under-dosing. This study was designed to define the role of IV busulfan in different transplant and disease settings. PATIENTS AND METHODS: The study included 43 patients with various hematologic malignancies conditioned with high-dose IV busulfan-containing regimens prior to allogeneic transplantation. The donors were HLA-matched siblings (n=24), matched unrelated (n=14), or 1-antigen mismatched related donors (n=5). Outcome parameters were recorded. RESULTS: Forty-two patients had initial engraftment. The toxicity profile was favorable. No patient developed veno-occlusive disease of the liver. Acute graft-vs-host disease (GVHD) grades II-IV occurred in 18 patients (42%). Six patients died of treatment-related causes, five of complications related to acute GVHD, and only one died of organ toxicity. Actuarial non-relapse-related mortality risk was 10% at day 100 and 18% at 2 years posttransplant. The actuarial 2-year overall survival and disease-free survival (DFS) rates were 63% and 44%, respectively. Disease status other than refractory relapse, myeloid disease, and no severe GVHD posttransplant predicted for longer DFS in a multivariant model. CONCLUSIONS: IV busulfan-containing regimens allow consistent engraftment of allografts from related and unrelated donors such that myeloablation is administered with a toxicity profile typical of non myeloablative conditioning. Favorable outcome was seen in patients with myeloid leukemias and in early or intermediately advanced disease; however, this regimen may not be sufficiently cytoreductive in patients with very advanced or active leukemia and in acute lymphoblastic leukemia. IV busulfan merits further study to better define its role as a preferred substitute for oral busulfan in pretransplant conditioning. PMID- 12763144 TI - [Assessment of the use of transfusion therapy and complications in orthopedic surgery in patients with sickle-cell anemia: retrospective study]. AB - Red blood cells (RBCs) transfusion is a common practice in the treatment or for the prevention of complications of patients with sickle-cell disease. In surgery, pre-operative transfusions are frequently given to prevent peri-operative complications. There is no consensus however on the best regimen of transfusion for this purpose. The transfusion techniques are muliple. In addition, pre operative transfusion therapy is reported to be largely responsible for an increased morbidity and mortality in patients with sickle cell anemia undergoing surgery. During the period 1990-2000, 16 patients (4 men and 12 women) with a mean age of 37 years and various major sickle cell hemoglobinopathies underwent 32 total hip arthroplasty for femoral head necrosis. Nine patients with sickle cell trait were included as control group. Twelve of them had haemoglobin SS (HbSS), 2/16 had HbSC, 2/16 had HbS/betathalassemia. Operative transfusion were given in only 12/32 procedures, 4 were performed pre-operatively and 8 intra operatively. Simple transfusion (mean: 2.5 packed red cells) were administered in all the procedures. The main complications observed in our patients were anemia by hemolysis and haemorrhagic shock, vaso-occlusive crisis and chest syndrome. Anemia requiring transfusions was significatively related to the procedures with pre-operative transfusion. In the light of our result, we would like to propose transfusional protocol--if needed--only intra-operatively. PMID- 12763146 TI - [NAT and transfusion safety: BioMerieux/Roche production chain performances- Chiron production chain performances]. PMID- 12763147 TI - [Measuring the HIV viral load with LCx (Abbott): importance for the therapeutic follow-up of 3 patients infected by type O HIV]. PMID- 12763149 TI - [Recent HCV infections in blood donors (1994-2001)]. PMID- 12763148 TI - [Antigen and viral load]. PMID- 12763150 TI - [Viral safety in multi-organ harvesting: the point of view of the virologist]. PMID- 12763151 TI - [Papillomavirus screening: shall it be performed systematically?]. PMID- 12763152 TI - [Prospects in blood transfusion]. AB - What will be the evolution of blood transfusion in the next 10 years? What are the scientific and medical arguments to help the decision makers to propose the developments? Many scientific and clinical studies show that blood substitutes are not ready for use in man. So, for a long time, blood collection in man will still be a necessity to prepare cell concentrates (red blood cells and platelets) and fresh frozen plasma. During this period, blood safety will be based on development of testing technics and preparation processes of blood products. Another major point will be a better clinical use of blood derivates. Cellular therapy will be probably only a way of diversification in blood transfusion centers in partnership with hospitals. PMID- 12763153 TI - [Self assessment tools]. AB - Self assessment tools are an integral part of quality process and are involved in developing a quality assurance program. Improvements is becoming a necessity for the new international standards Iso 9001. After the evaluation of the situation and the measure of the results, the improvement developing program results from self assessment. This article intends to provide a better understanding of self assessment processing. The first part presents the technique used for the elaboration of the questionnaire in relation with 'good transfusion practices' and the tools used to identify opportunities for quality improvement. The involvement of the whole staff is the key for the efficiency of appropriate action. The second part presents how these tools can be applied in blood transfusion organisation and in some specific applications. PMID- 12763154 TI - Influence of GnRH administration on timing of the LH surge and ovulation in dwarf goats. AB - This study was conducted to determine whether or not exogenous gonadotropin releasing hormone (GnRH) alters the timing or improves the synchrony of estrus, the LH surge, and ovulation following estrous synchronization in dwarf goats, and to assess the effects of season on these parameters. In January and June, estrus was synchronized in 12 Pygmy and Nigerian Dwarf goats with a 10-day progestagen sponge, 125 microg cloprostenol i.m. 48 h before sponge removal, and 300 IU equine chorionic gonadotrophin (eCG) i.m. at sponge removal. Six of the 12 goats were given 50 microg GnRH i.m. 24h after sponge removal. Onset of estrus was monitored using two males. Samples for plasma LH were collected at 2 h intervals beginning 22 h after sponge removal and ending at 48 h in January and at 58 h in June. Time of ovulation time was confirmed by laparoscopy at 36, 50, 60, and 74 h in January and at 50, 60, and 74 h in June. Administration of GnRH had no significant effect on the onset of estrus; however, it reduced the interval from sponge removal to the LH surge and improved the synchrony of the LH surge (P<0.05). Treatment with GnRH also reduced the interval from sponge removal to ovulation and improved the synchrony of ovulation (P<0.05). Season had a significant effect on the timing and the synchrony of estrus with and without GnRH treatment (P<0.05). A seasonal shift was also observed in the timing of the LH surge in the absence of GnRH treatment (P<0.05). Further research is required to determine the optimum time for GnRH administration and the minimum effective dose in dwarf goats. PMID- 12763155 TI - Use of G1.2/G2.2 media for commercial bovine embryo culture: equivalent development and pregnancy rates compared to co-culture. AB - The expanded application of commercial bovine IVM, IVF, and IVC systems is dependent on the ability to produce embryos in culture that are capable of producing normal pregnancies. Because serum containing culture systems can induce neonatal and fetal problems there exists a definite need for a serum-free culture system that produces viable blastocysts. This study demonstrated that the physiological sequential media system G1.2/G2.2 could produce bovine blastocysts at rates equivalent to co-culture. Additionally, these blastocysts had equivalent or increased cell numbers and inner cell mass development. Blastocysts produced in the G1.2/G2.2 culture system produced pregnancies following both fresh transfer and cryopreservation at equivalent rates to co-culture. Finally, this study demonstrated that the media system G1.2/G2.2 could be used in a commercial OPU transfer program without any loss in the numbers of blastocysts produced or the numbers of pregnancies resulting following transfer from either fresh or cryopreserved blastocysts. PMID- 12763156 TI - Effect of dialysis on quality characteristics of turkey semen during liquid storage. AB - Low molecular weight substances such as zinc and peroxides are present in seminal plasma and are responsible for deleterious effects in stored semen. On the contrary, molecules larger than 50 kDa are beneficial to in-vitro storage of spermatozoa. Since the effects of different seminal plasma fractions in turkey semen are not completely known, the purpose of the study was to determine the effects of turkey semen dialysis with a 12-14 kDa cut-off on viability, hypo osmotic membrane integrity, or sperm motility of turkey spermatozoa stored up to 48 h at 5 degrees C. Twelve pools of semen, each pool originating from four toms, were used. Each pool was divided into two aliquots, one of which was dialyzed while the other represented the control. Each semen aliquot was evaluated for sperm viability, membrane integrity and motility after 6, 24 and 48 h of in-vitro storage. Cold storage of turkey semen for 48 h significantly worsened (P<0.01) sperm viability, hypo-osmotic membrane integrity, and sperm motility index of both control and dialyzed samples. After 24 and 48 h sperm viability, membrane integrity and sperm motility index were better (P<0.01) in dialyzed semen compared to the control. PMID- 12763157 TI - Stereological characterization of bovine (Bos taurus) cumulus-oocyte complexes aspirated from small antral follicles during the metestrous and proestrous phases. AB - Bovine cumulus-oocyte complexes (COCs) aspirated from slaughterhouse ovaries are used for in vitro maturation and fertilization after selection on the basis of morphological appearance of the cumulus and ooplasm. In this context, a quantitative characterization of COCs could provide additional criteria for selecting the most competent complexes. Bovine COCs from small (1-4mm) antral follicles were aspirated from metestrous and proestrous stage ovaries of Holstein Friesian cows, fixed in glutaraldehyde, randomly embedded in glycol-methacrylate, and sectioned at 20 microm. The unbiased nucleator principle of stereology was used for estimating the mean volumes of complexes, oocytes, cumulus cells, and nuclei of oocytes and cumulus cells. The thickness of the zona pellucida and the relative numerical percentages of the several morphological types (C1, C2 and C3) of cumulus cells were also evaluated. The optical dissector procedure was used for cumulus cell sampling. Quantitative data show that the variability among complexes is generally high, especially for the volume of COCs. There were no linear correlations between the studied parameters, except between the volume of the oocyte and nucleus at metestrus. At proestrus, the volumes of COCs, oocytes and nuclei of oocytes, the volume of follicular cells and the thickness of the inner zona pellucida, were significantly higher than at metestrus. The relative numerical frequency of follicular type C1 cells was lower whereas that of type C3 cells was higher at proestrus than at metestrus. In conclusion, small antral follicles had larger COCs and oocytes at proestrus compared to metestrus and the COCs also had a higher percentage of follicular type C3 cells. Results suggest that for the same type of follicle size there may exist different functional populations of COCs at distinct stages of the bovine ovarian cycle. PMID- 12763158 TI - Double vitrification of rat embryos at different developmental stages using an identical protocol. AB - The aim of the present investigation was to test the effectiveness of a method of vitrifying rat embryos at different stages of development (from early morula to expanding blastocyst) in a double vitrification procedure. Wistar rat embryos were vitrified and warmed in super-fine open-pulled straws (SOPS). Before being plunged into liquid nitrogen, the embryos were exposed to 40% ethylene glycol+0.75 M sucrose in TCM-199+20% fetal calf serum (FCS) for 20s at 38 degrees C. Subsequent warming and direct rehydration of the embryos was conducted in culture medium (TCM-199+20% FCS) at 38 degrees C. Early morula stage (7-10 blastomeres) embryos (n=358) were vitrified, warmed and cultured in vitro (EM group). Batches of these embryos were then cryopreserved again (revitrified) at the early blastocyst (EB group, n=87), blastocyst (B group, n=93) or expanding blastocyst stage (ExpB group, n=73). After the first (EM group) and repeated (EB, B, and ExpB groups) vitrification procedures, developmental rates of 81, 83, 34 and 76%, respectively were achieved (for EM-EB-ExpB P>0.1; for EM, EB, ExpB-B P<0.005). Our data demonstrate the possibility of using the described identical protocol for the SOPS vitrification of rat early morulae, early blastocysts and expanding blastocysts. The low survival rate of blastocysts subjected to double vitrification requires further investigation. PMID- 12763159 TI - Seasonal changes in semen quality and freezability in the Warmblood stallion. AB - The objective of this study was to investigate seasonal changes in stallion semen quality and to determine the best time for semen cryopreservation. Experiments were performed using 10 Warmblood stallions from the National Stud Farm in Avenches (Switzerland). Ejaculates were collected and frozen every other week during 1 year from January to December 1999. Volume, concentration, and motility, and the number of morphologically normal sperm and sperm with major defects (abnormal heads, acrosome defects, nuclear vacuoles, proximal droplets, abnormal midpieces) were evaluated. For all frozen-thawed semen samples motility as well as viability (SYBR-14/PI) was tested, and the hypoosmotic swelling test (HOS) was performed. To analyze seasonal differences 4 periods of 3 months each were defined: autumn (September, October, November), winter (December, January, February), spring (March, April, May) and summer (June, July, August). During the 1 year experiment all semen quality parameters showed a clear seasonal pattern. The volume, total sperm count and motility in fresh semen were significantly higher (P<0.05) in summer than in winter, while sperm concentration was significantly lower in summer compared to the other seasons. Regarding morphology, normal sperm was significantly lower (P<0.05) in summer than at any other time of the year and higher values (P<0.05) were found for major defects in summer than in spring and autumn. In frozen-thawed semen motility was significantly (P<0.05) improved in autumn when compared to spring and summer. Viability was lowest in summer and differed significantly (P<0.05) from other seasons. The HOS test revealed significantly more (P<0.05) membrane damaged spermatozoa in winter than in spring, summer and autumn. Our results demonstrate that in our climatic conditions clear seasonal differences occur in semen quality of fresh and frozen-thawed semen and that cryopreservation of stallion semen should preferably be performed in autumn. PMID- 12763160 TI - Dimethyl sulfoxide influx in turbot embryos exposed to a vitrification protocol. AB - The particular characteristics of fish embryos require the development of specific methods for cryopreservation. One of the main obstacles is related to the presence of membranes and compartments with different water and cryoprotectant permeability. To assess dimethyl sulfoxide (Me2SO4) permeability, we exposed turbot embryos (Scophthalmus maximus) at F stage (tail bud) to the cryoprotectant solutions used in a vitrification protocol and then evaluated the Me2SO4 content inside the embryo using high-performance liquid chromatography (HPLC). The Me2SO4 influx was analyzed in normal embryos and in embryos treated with pronase (2mg/ml) in order to increase chorion permeability. The evaluation was made after each step of cryoprotectant incorporation and removal. Three embryo compartments were distinguished: the perivitelline space (PVS), the yolk sac (YS) and the cellular compartment (CC), and the relative volumes of each, estimated using stereoscopic microscopy imaging, were 11.37, 81.23 and 7.40%, respectively. The Me2SO4 concentration inside the embryos was calculated based on their entrance into one, two or three compartments. Results suggest high entrance of Me2SO4 into the PVS and a low concentration of this cryoprotectant inside the other compartments. Pronase did not significantly increase Me2SO4 influx, but facilitated its elimination during the washing steps. PMID- 12763161 TI - Conception rates of dairy cows following early not-pregnant diagnosis by ultrasonography and subsequent treatments with shortened Ovsynch protocol. AB - Our objective was to determine the feasibility of prompt reinsemination of dairy cows when diagnosed not pregnant 27-29 days after first-service timed AI (TAI). We assumed that a first-wave dominant follicle was present at that time that would ovulate in response to GnRH once precocious luteal regression was induced after administration of PGF(2alpha). Cows that had not been detected in estrus and reinseminated by Days 27-29 after a first-service TAI were diagnosed not pregnant by ultrasonography. Nonpregnant cows from three herds were assigned randomly to receive either no further treatment until reinsemination (controls; n=189); 25mg i.m. of PGF(2alpha) and then reinsemination according to detected estrus (81 of 108) or at 72-80h after PGF(2alpha) treatment (PGF) in the absence of estrus (27 of 108); or 25mg i.m. of PGF(2alpha) followed by 100 microg i.m. of GnRH 48h later (PGF+GnRH) and then reinsemination after detection of estrus (9 of 160) or at 16-20h after GnRH (151 of 160). Blood samples were collected at the time of the not-pregnant diagnosis and again 48h later. Concentrations of progesterone before treatment with PGF(2alpha) were elevated (<1ng/ml) in 61% of the cows when PGF(2alpha) was administered and 81% of the cows given PGF(2alpha) had low (<1ng/ml) concentrations of progesterone 48h after PGF(2alpha). Treated cows were re-inseminated earlier (P<0.01; 31+/-1days) after first-service TAI than controls (55+/-1days). Conception rates after treatment were not different among treatments: PGF (22%), PGF+GnRH (23%), and control (23%). Average intervals from calving to conception were 22-23 days less (P<0.001) in treated cows than in controls. We concluded that treating nonpregnant cows with PGF(2alpha) on Days 27 29 after insemination produced acceptable conception rates when inseminations were made after detected estrus or when TAI was used after GnRH treatment. Further, both treatments reduced days between first-service TAI and second inseminations, and days from calving to conception. PMID- 12763163 TI - Use of high-resolution transrectal ultrasonography to assess changes in numbers of small ovarian antral follicles and their relationships to the emergence of follicular waves in cyclic ewes. AB - Transrectal ovarian ultrasonographic studies have shown that, in cattle, follicular wave emergence is associated with a large increase in the number of small antral follicles (4-6mm in diameter); an analogous association has not been found for small follicles (2-3mm in diameter) in the ewe. In previous studies in ewes, accurate assessment of the number of follicles has been limited to follicles > or =2 or 3mm in size. Newer, high-resolution equipment allowed us to identify follicles > or =0.4mm and to quantify all antral follicles > or =1mm in diameter in seven cyclic Western White Face ewes. This allowed us to expand the small follicle pool examined, from 1 to 4 follicles/day (2-3.5mm in diameter) in earlier studies, to 8-18 follicles/day (1-3mm in diameter). Total number of small follicles (> or =1 and < or =3mm in diameter) increased between Days -1 and 0 (Day 0=day of ovulation), and declined between Days 1 and 3 (P<0.05). There were no significant changes in the number of small or medium (4mm in diameter) follicles around days of follicle wave emergence (+/-2 days). The 1-3 follicles in the 2-3mm size range, which constituted a follicle wave (i.e. grew to > or =5mm in size before regression or ovulation), were the only small follicles to emerge in an orderly succession during the estrous cycle, approximately every 3-5 days. Thus, unlike in cattle, there is no apparent increase in numbers of small follicles at follicle wave emergence in cyclic sheep, and little evidence for selection of recruited follicles and follicular dominance. PMID- 12763162 TI - Hormonal dynamics and follicular turnover in prepuberal Mediterranean Italian buffaloes (Bubalus bubalis). AB - The aim of this study was the investigation of hormonal and ovarian follicular dynamics in prepuberal buffaloes (Bubalus bubalis) bred in Italy. Eleven 5-9 month old buffalo calves ranging in weight from 122 to 270kg, maintained under controlled nutritional and environmental conditions, underwent 50 days of ultrasonographic ovarian follicular monitoring in the months of October-December. Blood sampling for E(2) and FSH determination and ultrasonographic monitoring using a 7.5MHz linear probe and an ALOKA SSD-500 monitor were performed daily. No differences in any of the parameters under study were highlighted when calves were divided into two weight categories (<200 and >200kg) and thus data were pooled. In this study, values are reported as mean+/-S.D. A range of two-six regular follicular waves was reported among calves with an average of 4+/-1.1. Overall interval (days) between wave emergence was 9.9+/-2.8 and largest diameters (mm) of dominant and first subordinate follicles were 8.4+/-1.2 and 4.8+/-0.6, respectively (P<0.05). With the exception of one calf, some minor follicular waves (short waves or SWs; 1.6+/-1), lasting <10 days (6.1+/-1.2) were reported. They were monitored contemporaneously on the ovary contralateral (n=7) or ipsilateral (n=3) to the main follicular wave. Growth rate (mm per day) of dominant follicles (DF) was significantly faster than for corresponding subordinate follicles (SF) and follicles of SWs (1.08+/-0.2 versus 0.79+/-0.1 and 0.83+/-0.1, respectively, P<0.05). The static phase (days) lasted longer in DF compared to SF and SW (5.4+/-1.8 versus 2.4+/-1.2 and 2.6+/-1, respectively, P<0.05). The regressing phase (mm per day) was similar among DF, SF and SW (0.86+/-0.2, 0.94+/-0.2 and 0.84+/-0.1, respectively, P=0.09). Episodic spikes of E(2) and FSH were reported, corresponding to wave development throughout the course of investigation. In conclusion, the majority of buffalo calves displayed a typical pattern of regular follicular development in conjunction with a dynamic trend of ovarian and hypophyseal hormones. Some minor follicle turnover was reported with parallel main follicular waves. PMID- 12763164 TI - Effect of thawing procedure on cryosurvival of deer spermatozoa: work in progress. AB - The objective of this study was to evaluate the effect of the thawing procedure on deer semen freezability. Frozen semen from the Genetic Resource Bank (GRB) of the Zoological Park of Buenos Aires (Argentina) was used. Seven mature stags (two red deer, two Pere David's deer and three fallow deer) were used as semen donors. Semen was diluted with a TRIS-egg yolk medium, packed in 0.25 ml straws and frozen in nitrogen vapour. For thawing, the frozen straws were subjected to the following procedures: (I) 70 degrees C, 5s; (II) 50 degrees C, 8s and (III) 37 degrees C, 10s. Freeze-thaw motility percentage (FMP) and spermatozoa rating (FMR) were determined subjectively. Viability and acrosome integrity (NAR) were also assessed and the hypo-osmotic swelling test (HOST) was used to assess membrane integrity. Freeze-thaw motility percentage, FMR and NAR were assessed after an incubation of 1h in citrate-yolk at 42 degrees C, and FMP and FMR after 2h of incubation under the same conditions. The thawing procedure did not have an effect on the seminal characteristics evaluated immediately after this process. However, differences in FMP after 2h of incubation (P<0.05) were found between the procedures, with the best overall recovery rates after freezing and thawing found with the use of protocols II (intermediate thawing) and III (slow thawing). Therefore, thawing protocols II and III, those that provide intermediate and slow thawing rates, were the most beneficial for semen thawing of the different cervid species analysed in this study. PMID- 12763165 TI - Plasma progesterone profiles, ovulation rate, donor embryo yield and recipient embryo survival in native Saloia sheep in the fall and spring breeding seasons. AB - The response to superovulatory (SOV) and estrus synchronization (ES) treatments and the fertility of donor (n=68) and recipient (n=118) Saloia ewes was evaluated in the fall and spring breeding seasons. The proportion of acyclic ewes at treatment time was significantly higher in the spring than in the fall (42.6% versus 4.0%, P<0.00001). Donors treated with eCG had a significantly higher mean number of follicles over 5mm in diameter in the ovaries at embryo recovery and a significantly lower mean efficiency of recovery than FSH-treated ewes. These negative effects were more pronounced in the fall than in the spring, which resulted in a significantly lower mean number of total and fertilized ova recovered from eCG-treated ewes, compared to FSH donors in the fall, but not in the spring. Season had no significant effect on the ovulation rate and plasma P4 concentrations of recipients treated with a progestagen plus eCG combination. Although the recipient lambing and embryo survival rates were higher in the fall than in the spring the differences were not significant. No significant differences were observed in the ovulation rate or P4 concentrations of recipients that lambed compared to those that did not lamb. These preliminary results show that, in Portugal, response of Saloia ewes to SOV or ES treatments and donor fertility following the SOV treatment were similar in the spring and the fall, which suggests that in the spring acyclic ewes are in moderate anestrus. The effect of season on fertility following embryo transfer should be confirmed in further studies involving a larger number of animals. The semilaparoscopic transfer method reported here allowed lambing and embryo survival rates higher (although not significantly) than a standard surgical approach. PMID- 12763166 TI - Comparison of proteins synthesized by polarized caprine oviductal epithelial cells and oviductal explants in vitro. AB - Our objectives were to compare proteins secreted by caprine oviductal explants and oviductal epithelial (OE) cells in vitro. Oviducts were collected from goats on Days 1 (n=5) and 5 (n=5) of the estrous cycle. Radiolabeled secretory proteins from tissue segments and cell cultures were visualized using SDS-PAGE and fluorography. After culture, media from ampulla oviduct segments collected on Days 1 and 5 of the estrous cycle contained an acidic 97 kDa protein, which was greatly reduced in culture medium obtained from infundibulum and isthmus oviduct segments. A complex of low molecular weight proteins (14-26 kDa) could be modulated by estradiol when OE cells were cultured on plastic. This complex was constitutively expressed when OE cells were cultured on Matrigel-coated filters. Polarized OE cells were also capable of compartment-specific secretion of [L (35)S]-methionine-labeled proteins. A 45 kDa acidic protein was predominantly secreted into the apical compartment while a 66 kDa acidic protein was preferentially localized in the basal compartment. Proteins secreted by OE cells were similar to proteins secreted by tissue segments in vitro. Therefore, under well-defined culture conditions OE cells may be useful in enhancing in vitro fertilization or early embryonic development. PMID- 12763167 TI - Effects of presynchronization during the preservice period on subsequent ovarian activity in lactating dairy cows. AB - Among the strategies aimed at overcoming difficulties in estrus detection in dairy herds, presynchronization with two PGF2alpha treatments 14 days apart before a timed AI protocol has been related to a significant increase in pregnancy rates. The aim of the present study was to evaluate the effects of presynchronization during the preservice period on subsequent ovarian activity in clinically normal lactating dairy cows. A second objective was to evaluate the incidence of reproductive disorders on Day 50 postpartum. Depending on the chronological order of parturition, cows were alternately assigned to a control (n=102) or treatment (n=101) group. Animals in the treatment group were administered two cloprostenol treatments 14 days apart, beginning on Day 22 postpartum. The reproductive tract of each animal was examined ultrasonographically on Days 43 and 50 postpartum to monitor ovarian structures and uterine contents. Blood samples were collected on Day 50 for progesterone determination. Cows were inspected for signs of estrus between Days 50 and 71 postpartum and were then inseminated. Follicular persistence rates were similar in the presynchronized (14.9%) and control (13.7%) groups. Cows in the presynchronized group showed a lower metritis-pyometra rate (0% < 3.9%; P=0.045); a lower ovarian cyst rate (3% < 10.8%; P=0.03); a higher luteal activity rate (progesterone concentrations>/=1ng/ml) on Day 50 postpartum (76.2% > 52.9%; P=0.0005); a higher estrus detection rate (73.3% > 47.1%; P<0.0001); a higher ovulation rate (72% > 44%; P<0.0001) and a higher pregnancy rate (29.7% > 15.7%; P=0.02) than controls. Our results indicate that presynchronization during the preservice period reduces the incidence of ovarian cysts and metritis-pyometra determined on Day 50, and improves ovarian activity from Days 50 to 71 postpartum along with pregnancy rates in clinically normal lactating dairy cows. PMID- 12763168 TI - Semen quality and presence of cytokines in seminal fluid of bull ejaculates. AB - It has been postulated that cytokines play an immunoregulatory role in human semen. The presence and levels of various cytokines, namely the interleukins 6, 10 (IL-6 and IL-10) and tumor necrosis factor alpha (TNFalpha) were investigated in the seminal plasma of six fertile crossbred bulls (Bos taurus x Bos indicus), using specific enzyme immunoassay. Cytokine levels were related to the semen quality of these bulls. A direct significant correlation between the level of IL 10 and individual sperm motility (r=0.84, P<0.036) was observed. The levels of IL 6 and IL-10 were inversely correlated (r=-0.93, P<0.006) and likewise in the same way an inverse correlation between the level of IL-6 and TNFalpha was observed (r=-0.84, P<0.037). These results confirm that cytokines are present in bull semen as has been demonstrated in human semen. PMID- 12763169 TI - Synergistic action of growth hormone and insulin-like growth factor I (IGF-I) on proliferation and estradiol secretion in porcine granulosa and theca cells cultured alone or in coculture. AB - The effect of growth hormone (GH) on insulin-like growth factor I (IGF-I) secretion and the effects of GH and IGF-I on [(3)H] thymidine incorporation and estradiol (E2) secretion by theca interna (Tc) and granulosa cells (Gc) cultured alone and in coculture were studied in cultured porcine follicular cells, prepared from small (SF), medium (MF) and large (LF) preovulatory follicles. We demonstrated that both Tc and Gc secrete IGF-I and that GH had no effect on IGF-I secretion by Tc but, increased IGF-I secretion by Gc isolated from SF and cultured alone or in coculture. IGF-I stimulated secretion of E2 by all cells, except in Tc derived from SF in which the effect was not statistically significant. The only stimulatory effect of concurrent treatment with GH on E2 secretion was noted in Tc derived from MF. IGF-I increased the [(3)H] thymidine incorporation in all Tc cells but GH did not augment this effect. In Gc, IGF-I stimulated [(3)H] thymidine incorporation in cells derived from SF and MF but not from LF. GH had no stimulatory effect except on Gc derived from LF and grown alone. The highest stimulatory effect was observed in SF. This was smaller in MF and no effect was noted in LF. In conclusion, our work shows that both Gc and Tc are sites of IGF-I production and for the first time shows the stimulatory role of IGF-I in proliferation of Tc cells derived from all types of follicles and augmentation of E2 secretion in Tc derived from MF and LF. The promotion of the mitogenic activity in Tc by IGF-I during all stages of follicular development suggests an important role for theca cells in follicular growth. PMID- 12763170 TI - Chromosome abnormalities in secondary pig oocytes matured in vitro. AB - Abnormalities of chromosome segregation during in vitro maturation of oocytes cause failure of in vitro fertilization. Oocytes collected from pig ovaries after slaughter were matured in vitro (IVM) for 30-48 h. In total, 1144 secondary oocytes were studied cytogenetically. An unreduced (diploid) chromosome set was identified in 146 spreads (12.8 %). A higher proportion of diploidy was noticed in secondary oocytes matured for 40 h and longer (15.0 %) than in the groups matured for 30 and 36 h (9.0 %). Among 998 secondary oocytes with the reduced chromosome number, 612 could be analyzed in detail. Hypohaploidy (n=19-1) was identified in 22 cells (3.59 %) and a hyperhaploid (n=19+1) set of chromosomes was identified in 15 cells (2.45 %). The rate of aneuploidy, estimated by doubling the rate of hyperhaploidy was 4.9 %. It was also found that aneuploid spreads occurred more frequently in the group of oocytes matured for 40 h and longer. Small acrocentrics were mostly found as an extra chromosome in the hyperhaploid spreads. Our study indicates that to avoid an excess of chromosomally abnormal secondary oocytes, IVM duration of pig oocytes should not exceed 40 h. PMID- 12763171 TI - Luteal activity at the onset of a timed insemination protocol affects reproductive outcome in early postpartum dairy cows. AB - This study was designed to compare two timed insemination protocols, in which progesterone, GnRH and PGF2alpha were combined, with the Ovsynch protocol in presynchronized, early postpartum dairy cows. Reproductive performance was also evaluated according to whether cows showed high or low plasma progesterone concentration, at the onset of treatment. One hundred and six early postpartum dairy cows were presynchronized with two cloprostenol treatments given 14 days apart, and then assigned to one of the three treatment groups. Treatments for the synchronization of estrus in all three groups started 7 days after the second cloprostenol injection, which was considered Day 0 of the actual treatment regime. Cows in the control group (Ovsynch, n=30) were treated with GnRH on Day 0, PGF2alpha on Day 7, and were given a second dose of GnRH 32 h later. Cows in group PRID (n=45) were fitted with a progesterone releasing intravaginal device (PRID) for 9 days, and were given GnRH at the time of PRID insertion and PGF2alpha on Day 7. In group PRID/GnRH (n=31), cows received the same treatment as in the PRID group, but were given an additional GnRH injection 36 h after PRID removal. Cows were inseminated 16-20 h after the administration of the second GnRH dose in the Ovsynch group, and 56 h after PRID removal in the PRID and PRID/GnRH groups. Ovulation rate was determined on Day 11 postinsemination by detecting the presence of a corpus luteum in the ovaries. Lactation number, milk production, body condition at the onset of treatment and treatment regime were included as potential factors influencing ovulation and pregnancy after synchronization. Logistic regression analysis for cows with high and low progesterone concentration on treatment Day 0 revealed that none of the factors included in the models, except the interaction between progesterone and treatment regime, influenced the risk of ovulation and pregnancy significantly. In cows with high progesterone concentration at treatment onset, Ovsynch treatment resulted in a significantly improved pregnancy rate over values obtained following PRID or PRID/GnRH treatment. In cows with low progesterone concentration, PRID or PRID/GnRH treatment led to markedly increased ovulation and pregnancy rates with respect to Ovsynch treatment. These findings suggest the importance of establishing ovarian status in early postpartum dairy cows before starting a timed AI protocol, in terms of luteal activity assessed by blood progesterone. PMID- 12763172 TI - [Medical-psychiatric units, a concept to be developed]. PMID- 12763173 TI - [Heterozygous prothrombin gene mutation G20210A and associated diseases]. AB - PURPOSE: Prothrombin gene mutation G20210A (factor II) is, in frequency, the second genetic polymorphism involved in venous thrombosis. We report a retrospective studies on 38 patients issued from our medical department, all heterozygous for the factor II mutation and a literature review. METHODS: We have studied 38 patients, all heterozygous for the factor II mutation, selected through a population of 516 tested patients issued from our medical department from 1997 to 2002. The research was performed face with history of thrombotic or obstetrical events, angiopathy or familial screening. RESULTS: Twenty out of thirty-eight patients have at least one episode of venous thrombosis: superficial thromboses, deep thromboses and/or pulmonary embolism. One case of cerebral thrombophlebitis is observed. Venous thrombotic risk factors are associated in 12 cases (60%). Four out of thirty-eight patients have one episode of arterial thrombosis: cardiovascular, peripheral or cerebral. Arterial thrombotic risk factors are associated in all cases. Median age of the first venous thrombosis is earlier than the one of arterial thrombosis (39.11 versus 49.25 years). CONCLUSION: Our studies confirms the interest to search the prothrombin gene mutation when faced with a venous thrombotic event (deep vein thrombosis and/or pulmonary embolism) with or without acquired risk factors. Its involvement in thrombotic arterial disease is still a matter of debate. Data concerning its involvement in systemic diseases and angiopathies (thromboangeitis obliterans, Raynaud's phenomenon and migraine) are still needed. Mechanisms of thromboses could be an increase of prothrombin plasma level with high thrombin synthesis. PMID- 12763175 TI - [Coagulation inhibitors in severe sepsis: state of the art]. AB - OBJECTIVE: To present and discuss the rationale and the results of clinical trials using supplementation with physiologic anticoagulants (Tissue Factor Pathway Inhibitor (TFPI), AntiThrombin (AT), and Protein C (PC) in patients with severe sepsis. RATIONALE: An early activation of the coagulation cascade occurs in severe sepsis. TFPI, AT, and PC are major inhibitors of the coagulation cascade, and additionally modulate inflammatory and vascular reactions. They are consumed or inhibited in the sepsis pathologic process. Therapeutic supplementation with these inhibitors could improve the sepsis-induced organ failures and mortality. CLINICAL RESULTS: Randomized controlled studies were recently completed. No effect on the mortality rate could be documented after treatment with recombinant TFPI. AT concentrates neither improve mortality, but a biological interaction with heparin therapy could have biased the study results. Treatment with recombinant activated PC (alpha-drotrecogin) was associated with a significant reduction in the mortality rate of severely ill patients and received recently the approval from FDA and EC authorities in this indication. An increase in the rate of hemorrhagic adverse effects has been observed with these compounds, justifying a strict observance of contraindications and of patients selection. PROSPECTIVE: Additional studies are needed to give confirmation of the positive effects of activated PC supplementation in less severely ill patients, children and specific clinical situations. The effects of new anticoagulant compounds are currently evaluated in preclinical studies. PMID- 12763174 TI - [Systemic lupus erythematosus with disease onset after age 65]. AB - OBJECTIVES: Systemic lupus erythematosus with disease-onset in the elderly has rarely been studied (only one report about 21 patients with disease onset at 65 and older). Is the management of this pathology modified in this population? METHODS: Seventeen hospitalised cases of lupus patients with disease onset at 65 or older are retrospectively reported between 1988 and 2000. The results are compared with those of younger subjects. RESULTS: The female to male ratio is 1.83. Mean age at disease onset is 71.9 +/- 3.5 years. Mean duration of follow-up is 3.5 +/- 2.4 years. Main initial symptoms are: deterioration of general status (41%), arthritis (35%), cutaneous manifestations (35%), thrombo-embolism (24%) and pleuritis (18%). Malar rash is uncommon (12%). Nephropathy is never a revealing symptom and is rarely serious during the disease's evolution. Like in neurologic manifestations, the etiology has to be discussed in relation to associated co-morbidities. Concerning haematologic features, lymphopenia is found in 82% of the cases with a questionable specificity. Antinuclear antibodies are constant, anti-dsDNA antibodies are found in 82% of the cases, antibodies to extractable nuclear antigens in 50%, and anticoagulant circulating activity in 59%. Prognosis is difficult to assess in such a limited series but 5-years survival probability is 83%. Glucocorticoid lead to 50% of major complications. CONCLUSIONS: This study focuses on the particular initial manifestations of systemic lupus erythematosus in the elderly (deterioration of general status, thrombosis, unusual cutaneous symptoms), and on the specificity of differential diagnosis and treatment. PMID- 12763176 TI - [Cyclines and acne: pay attention to adverse drug reactions! A recent literature review]. AB - PURPOSE: Cyclines are broad-spectrum antibiotics often used in acne. Side effects might occur precociously or more insidiously during long term treatment such in acne. The aim of this study is to review all adverse effects recently reported in this indication. CURRENT KNOWLEDGE AND KEY POINTS: A literature review from 1997 to 2001 has been conducted. Seventy-six articles reporting 250 cases have been found. Data analysis indicated that minocycline is the most widely incriminated molecule. Seventy-two cases revealed autoimmune disorders : lupus like syndrome associated or not with autoimmune hepatitis, 5 cases developed vasculitis. These reactions occurred in long term treatment (several weeks to several months). Fifteen cases of hypersensitivity syndromes and 3 cases of serum sickness like illness have been reported. These reactions occurred within the first weeks of exposure. Twenty-four cases of pseudotumor cerebri and 123 cases of abnormal pigmentations (skin, nails, mouth, bones or organs) have also been found. Eight cases presented other diseases. FUTURE PROSPECTS AND PROJECTS: Adverse effects of cyclines might be serious and sometimes unknown. Long term treatment by tetracyclines must be researched in patients presenting such symptoms. Moreover, several adverse drug reactions might be avoided by an optimal use of the drug (oesophageal ulcerations, photosensitivity) or by shorter periods of treatment (autoimmune disorders, pigmentations); only DRESS are drug adverse reactions unpredictable and sometimes severe. PMID- 12763177 TI - [Multiple pseudo-cellulitis plaques and Koplick's sign: a particular form of parvovirus B19 primo-infection in adults]. AB - INTRODUCTION: clinical presentation which seems to be more specific of this infection. EXEGESIS: A 35-year-old woman, 12 weeks pregnant, presented with a primary infection of parvovirus B19. The clinical presentation was characterized by pseudo-cellulitis plaques of the buttocks and the vulva, buccal enanthema with ulcerations and Koplick spot. CONCLUSION: This is the second observation which describes such cutaneous and mucosal manifestations associated with parvovirus B19 infection. This kind of clinical presentation should be systematically reported to become well known by the physicians as erythema infectiosum of fifth disease or "gloves and socks" syndrome. PMID- 12763178 TI - [Diagnosis of a metastatic valvular tumor in a living patient]. AB - INTRODUCTION: The discovery of an endocardial mass always raises the question of its nature. Infectious endocarditis is the most frequent cause, but others diagnoses must be considered. EXEGESIS: We report a case of endocardial metastasis originating from an upper respiratory tract epidermoid carcinoma in a 48-years-old man. The diagnosis was established while the patient was alive, and survival at the time of writing is 8 months. This case report provides an opportunity for discussion of the differential diagnosis when confronted with an endocardial tumor, i.e. bacterial endocarditis, non-bacterial thrombotic endocarditis, primary cardiac tumors, metastatic osteogenic sarcoma and Libman Sachs endocarditis. CONCLUSION: Non bacterial thrombotic endocarditis and valvular metastasis should be considered upon discovery of a valvular tumor, in the context of neoplastic disease. The prognosis of endocardial metastasis is poor, but early diagnosis and appropriate management should eventually prolong survival. PMID- 12763180 TI - [Cutaneous nodules to an aquarist]. PMID- 12763179 TI - [Sclerosing cholangitis as a complication of Langerhans'cell histiocytosis]. AB - INTRODUCTION: Langerhans'cell histiocytosis is a rare and mainly pediatric disease. Patients with hepatic involvement usually have a disseminated form of the disease, with poor prognosis. Sclerosing cholangitis can occur in 10 to 15% of multivisceral Langerhans'cell histiocytosis. We report the case of a 56-years old patient who developed sclerosing cholangitis 12 years after the diagnosis of Langerhans'cell histiocytosis. EXEGESIS: A 56-years-old man was admitted because of rapid general health impairment with epigastric pain. He was diagnosed as having Langerhans'cell histiocytosis 12 years ago because of a diabetes insipidus. Lungs were involved and during follow-up vertebral osteocondensation also developed. However, Langerhans'cell histiocytosis was clinically silent at the time of admission, without any treatment. Biologically, cholestasis and inflammation were noted. Digestive radiological investigations (echo-endoscopy, CT-scan, MRI) showed homogenous hepatomegaly and a diffuse infiltration of intra and extrahepatic bile ducts. Liver biopsy yielded the diagnosis of sclerosing cholangitis. Clinical and biological improvement occurred with oral corticosteroids (at 12 months after sclerosing cholangitis diagnosis). CONCLUSION: Sclerosing cholangitis is a potential complication of Langerhans'cell histiocytosis, mainly in its multivisceral form. It can occur at a median of 2 years after diagnosis in children, but occasionally much later in adults, whereas Langerhans'cell histiocytosis seems quiescent. Diagnosis is supported by radiological investigations and liver biopsy. As no drug therapy appears clearly effective, liver transplantation must frequently be considered in these patients. PMID- 12763181 TI - [Good syndrome and cytomegalovirus pneumonia]. PMID- 12763182 TI - [A very late onset ankylosing spondylitis]. PMID- 12763183 TI - [Hypothyroidism is sometimes a late complication after irradiation of the neck]. PMID- 12763184 TI - [Letter to the editor by X. Roblin et al., Influence of hypolipemic treatment on homocysteinemia]. PMID- 12763187 TI - Human brain regions required for the dividing and switching of attention between two features of a single object. AB - This combined PET and ERP study was designed to identify the brain regions activated in switching and divided attention between different features of a single object using matched sensory stimuli and motor response. The ERP data have previously been reported in this journal [64]. We now present the corresponding PET data. We identified partially overlapping neural networks with paradigms requiring the switching or dividing of attention between the elements of complex visual stimuli. Regions of activation were found in the prefrontal and temporal cortices and cerebellum. Each task resulted in different prefrontal cortical regions of activation lending support to the functional subspecialisation of the prefrontal and temporal cortices being based on the cognitive operations required rather than the stimuli themselves. PMID- 12763188 TI - Tactile-visual cross-modal shape matching: a functional MRI study. AB - The process and location of integration of information from different sensory modalities remains controversial. We used functional MRI to investigate the neural representation of cross-modal matching between tactile and visual shape information in eleven normal volunteers. During the scan, patterns of 2D shapes were presented both tactually and visually, simultaneously. Four different matching tasks were performed: tactile-tactile with eyes closed (TT), tactile tactile with visual input (TTv), visual-visual with tactile input (VVt), and tactile-visual (TV). The TT task activated the contralateral primary sensorimotor area, and the postcentral gyrus, superior parietal lobules, anterior portion of the intraparietal sulcus, secondary somatosensory cortex, thalamus, dorsal premotor area, cerebellum, and supplementary motor area bilaterally, without occipital involvement. Visual matching activated the primary visual cortex and the lingual and fusiform gyri bilaterally. A cross-modal area was identified by subtracting TTv images from TV images, subtracting VVt images from TV images, and then determining common active areas. There was one discrete area that was active bilaterally; the posterior intraparietal sulcus close to the parieto-occipital sulcus. These data suggest that shape information from different sensory modalities may be integrated in the posterior intraparietal sulcus during tactile visual matching tasks. PMID- 12763189 TI - Processing of normal, inverted, and scrambled faces in a patient with prosopagnosia: behavioural and eye tracking data. AB - In this study, we addressed the issue of a spared processing of faces in a patient (SB) with severe prosopagnosia. We designed an experiment comprising of two parts. In Part I, normal upright faces were entwined with scrambled faces, while in Part II normal upright faces were mixed with inverted faces, under unlimited time exposure. Performance, decision times, and eye movements were measured in both parts. The results indicated that SB categorised the normal faces better in the context of inverted faces than in the context of scrambled faces. Furthermore, SB's performance was better for the inverted faces than for the scrambled faces. Overall, SB performed better on the abnormal faces than on the normal faces, as did the control participants. Eye-tracking data showed that the pattern observed for the number of fixations and for exploration order was similar in SB and in controls. In the discussion, we propose that, despite his severe prosopagnosia, SB might have retained some kind of processing specific to face perception. Further investigations will be required, using limited time exposure, to determine the nature of this spared processing. PMID- 12763190 TI - Modulation of the N400 potential during auditory phonological/semantic interaction. AB - The processing of phonological and semantic word attributes has been commonly explored with electrophysiological methods using simple contexts where competition between features is eliminated. Conversely, the interaction between phonological and semantic attributes has not been systematically examined. We therefore recorded an event-related electrophysiological marker of word discordance, the N400, in response to sequences of auditory word pairs containing semantic incongruences, phonological discordances, or a mixture of the two. N400 enhancement to semantically unrelated words was systematically observed, whether the subjects heard the sequences passively (no instruction) or actively (semantic judgement task), and even in contexts where the task did not concern semantic attributes. In contrast, the N400 effect to phonologically unrelated (non rhyming) words was exclusively obtained in the active situation (phonological judgment), while it disappeared in passive conditions and during semantic/phonological interference. This suggests that the detection of semantic incongruences is a more robust and automatized mechanism than that of phonological ones, and tends to occlude this latter when both features are in competition. Our data also provide new elements supporting the persistence of the semantic N400 during 'shallow' word processing tasks, i.e. tasks that discourage analysis of semantic aspects of the words. PMID- 12763192 TI - Shared and dissociated cortical regions for object and letter processing. AB - The present study determined the extent to which object and letter recognition recruit similar or dissociated neural resources. Participants passively viewed and silently named line drawings of objects, single letters, and visual noise patterns and centrally fixated an asterisk. We used whole-brain functional MRI and a very conservative approach to hypothesis testing that distinguished among brain regions that were selectively activated by different experimental conditions and those that were conjointly activated. The left fusiform gyrus (BA 19 & 37) and left inferior frontal cortex BA(44/6) showed a greater degree of conjoined activation for objects and letters than selective activation for either category, whereas left inferior parietal cortex (BA 40) and the left insula showed a strong letter-selective response. Equal recruitment of left fusiform and inferior frontal regions by objects and letters reflects similar demands on cognitive processing by these two categories and argues against category-specific modules in these regions. However, cortical systems for object and letter processing are not completely shared given the exclusive activation of left inferior parietal cortex by letters. PMID- 12763191 TI - Modulation of neural responses to speech by directing attention to voices or verbal content. AB - We studied with functional neuroimaging the cortical response to auditory sentences, comparing two recognition tasks that either targeted the speaker's voice or the verbal content. The right anterior superior temporal sulcus responded during the voice but not during the verbal content task. This response was therefore specifically related to the analysis of nonverbal features of speech. However, the dissociation between verbal and nonverbal analysis was only partial. Left middle temporal regions previously implicated in semantic processing responded in both tasks. This indicates that implicit semantic processing occurred even when the task directed attention to nonverbal input analysis. The verbal task yielded greater bilateral activation in the fusiform/lingual region, presumably reflecting an implicit translation of auditory sentences into visual representations. This result confirms the participation of visual cortical regions in verbal analysis of speech. PMID- 12763193 TI - Development of isometric force and force control in children. AB - Fifty-six children between 5 and 12 years of age and 15 adults performed a task (pressing on a lever with the index finger of the preferred hand), in which a force had to be maintained constant at five levels with on-line visual feedback. Since this is a simple isometric task, the hypothesis is that optimal performance (in terms of force variability control) closely matches the maturation of the corticospinal tract up to age 10. It was found that maximum voluntary contraction (MVC) matured over the full range of the examined age groups. In contrast, the coefficient of variation of force showed maturation mainly up to the age of 9-10, as hypothesised. Gender differences were found for MVC but not for the other force control parameters. Power spectral density analysis revealed developmental differences in the lower (1-6 and 7-12 Hz) and higher frequencies bands (13-18 and 19-24 Hz). In the lowest frequency range the amount of energy decreased with age, presumably because young children (5-8 years of age) rely more heavily on control from proprioceptive and visual feedback. It is argued that with increasing neural maturation the control processes become more dependent upon internal representation manifested by feed forward control that starts to substitute feedback-based control. PMID- 12763194 TI - Distinct prefrontal cortex activity associated with item memory and source memory for visual shapes. AB - In contrast to item memory, which refers to recognition or recall of previously presented information, source memory refers to memory for the context of previously presented information. The relatively few functional MRI (fMRI) source memory studies conducted to date have provided evidence that item memory and source memory are associated with differential activity in right and left prefrontal cortex, respectively. To both confirm this distinction in prefrontal cortex and to determine whether other differences in the neural substrates associated with these cognitive functions exist, an event-related fMRI study was conducted. In this study, item memory and source memory encoding phases were identical; participants viewed a series of abstract visual shapes presented on the left or right side of the screen and were instructed to remember each shape and its spatial location. During the item memory retrieval phase, shapes from the encoding phase were intermixed with new shapes and participants made an old-new judgment. During the source memory retrieval phase, all shapes were from the encoding phase and participants made a left-right judgment. An event-related analysis of item memory and source memory revealed a right and left prefrontal cortex distinction. Moreover, only item memory was associated with activity in the medial temporal lobes. These results confirm and extend previous findings that item memory and source memory are associated with distinct neural substrates. PMID- 12763195 TI - The fate of sounds in conductors' brains: an ERP study. AB - Professional music conductors are required to home in on a particular musician but at the same time have to monitor the entire orchestra. It was hypothesized that this unique experience should be reflected by superior auditory spatial processing. Event-related brain potentials were obtained, while conductors, professional pianists, and non-musicians listened to sequences of bandpass filtered noise-bursts presented in random order from six speakers, three located in front and three to the right of the subjects. In different runs, subjects either attended the centermost or the most peripheral speaker in order to detect slightly deviant noise-bursts. For centrally located speakers, the ERPs showed a typical Nd attention effect for the relevant location with a steep decline for the neighboring speakers in all subject groups. For peripheral speakers, only the conductors showed attentional selectivity, while the Nd effect was of similar size for all three peripheral speakers in the other two groups. These ERP effects were paralleled by an enhanced behavioral selectivity in peripheral auditory space in conductors. Moreover, the pre-attentive monitoring of the entire auditory scene indexed by the mismatch negativity was superior in musicians compared to non-musicians. In conductors, the MMN was followed by a positivity suggesting an attention shift towards the deviant stimuli in this group only. PMID- 12763196 TI - Waking EEG power spectra in the rat: correlations with training performance. AB - Adult rats chronically implanted with supradural electrodes were telemetrically EEG recorded during a baseline session, a training session for a two-way active avoidance task, and a retention session. Rats were assigned to a fast learning (FL), slow learning (SL) and non learning (NL) group if they achieved criterion during the training session, the retention session, or in neither session. High resolution EEG analyses indicated that intergroup differences were present in the low frequency range of waking baseline power spectra. Moreover, baseline delta emissions directly correlated with freezings, and inversely correlated with avoidances, while emissions at 7-10 Hz directly correlated with avoidances and inversely correlated with freezings. Interestingly, during the first training period, waking delta emission selectively increased in FL rats in concomitance with a marked performance improvement; instead, SL and NL rats displayed increments at 7-9 Hz. In addition, freezings scored during the first two training periods directly correlated with post-training waking emission at 2 Hz, and inversely correlated with emission at 7-10 Hz. Conversely, escapes and avoidances directly correlated with waking emission at 7-10 Hz. The data indicate that (i) waking baseline power spectra differ among behavioral groups, and correlate with behavioral performance the following day; (ii) selective modifications of waking power spectra occur in each behavioral group during training; and (iii) behavioral responses during training correlate with post-training waking power spectra. Notably, the delta increment selectively occurring in training FL rats is assumed to reflect online memory processing leading to better performance. The latter observation supports the primary involvement of delta waves in learning. PMID- 12763197 TI - Striatal size, glucose metabolic rate, and verbal learning in normal aging. AB - The striatum has recently been implicated as an area that may mediate age associated cognitive decline because of diminution of volume and functional activity. We used 18F-fluorodeoxyglucose (FDG) with positron emission tomography (PET) and high-resolution magnetic resonance imaging (MRI) to examine the effects of age on striatal glucose metabolic rate and size in 70 healthy, normal subjects. During the FDG tracer uptake period, subjects performed a serial verbal learning task, based on the California Verbal Learning Test. PET images were co registered to the MR images. The interrelations among striatal glucose metabolic rate, size, and performance on the verbal learning task were examined with repeated-measures analysis of variance and correlational analysis. As age increased, relative glucose metabolic rate (GMR) increased in the putamen and decreased in the caudate. Female subjects had lower relative GMR than male subjects in the caudate, but equal in the putamen. Striatal size remained relatively constant across the lifespan in men but was lower in women aged 50-70 than in men. While there were significant associations between striatal activity and performance on the uptake task, these findings were mostly accounted for by age. The findings are consistent with our earlier report on the same cohort that demonstrated an age-related shift from anterior to posterior cortical metabolism, as the putamen receives primarily posterior cortical input and the caudate receives relatively more anterior cortical input. Findings of significant involvement of striatal functioning in verbal learning are most likely accounted for by age and suggest an age-related shift from anterior to posterior circuitry in the human telencephalon. PMID- 12763198 TI - Electrophysiological distinctions in processing conceptual relationships within simple sentences. AB - The aim of this study was to determine whether or not the brain distinguishes between two types of conceptual relationships between noun-phrases (NPs) and verbs during online processing of simple, unambiguous English sentences. A total of 15 participants read and made plausibility judgments on sentences that were presented word-by-word. Event-related potentials elicited by critical verbs were measured. In all cases, the critical verb assigned a thematic role of 'agent' to its subject NP. In non-violated sentences (e.g. "For breakfast the boys would only eat em leader"), the preceding NP was animate ("boys") and was a likely agent for a given verb ("eat") given its preceding context ("For breakfast"). In both types of conceptually violated sentences, the NPs were unlikely agents for the verbs given their preceding contexts. In 'thematic role animacy violations' (e.g. "For breakfast the eggs would only eat em leader"), the NP was inanimate ("eggs") and was therefore more likely to occupy the role of 'theme' than 'agent', i.e. eggs, being inanimate, cannot eat but they can be eaten. In 'non thematic role pragmatic violations' (e.g. "For breakfast the boys would only bury em leader"), the thematic role of agent assigned by the verb ("bury") to its preceding NP ("boys") is inherently acceptable (boys can bury), but the sentence is still pragmatically incongruous given the preceding context ("At breakfast"). As expected, the non-thematic role pragmatic violations elicited a significant N400 effect. The thematic role animacy violations elicited a smaller N400 effect that only approached significance across all participants. The thematic role animacy violations, however, elicited a significant P600 effect-an ERP component that is most commonly associated with processing syntactic information during language comprehension. We discuss the possibility that the P600 was elicited by the thematic role animacy violations (but not by the non-thematic role pragmatic violations) because, in the former but not the latter, there was an online attempt to structurally repair and make sense of the sentences by reassigning the thematic role of the NP that preceded the critical verb from 'agent' to 'theme'. Our findings suggest a qualitative neural distinction in processing these two types of conceptual anomalies within simple, unambiguous sentences. PMID- 12763199 TI - Processing of dynamic aspects of speech and non-speech stimuli: a whole-head magnetoencephalography study. AB - Clinical and experimental data indicate higher proficiency of the left hemisphere in encoding dynamic acoustic events such as rapid formant transitions (30-40 ms) that distinguish consonant-vowel syllables such as /ba/ or /da/. In order to further elucidate the underlying neurophysiological mechanisms, discrimination of /bi/-like formant transitions of variable duration (18, 36, 54, or 72 ms) from a steady-state /i/-like vowel was investigated by means of whole-head magnetoencephalography (MEG) both during visual distraction and selective attention. Voiced speech-like as well as unvoiced non-speech stimuli, matched for spectral envelope, served as test materials. Based on an oddball design, magnetic mismatch fields (MMF) were determined during an early (170-210 ms) and a late (230-290 ms) time window. Selective attention toward the deviant events resulted in enhanced MMFs particularly within the left hemisphere, indicating attention dependent left-lateralized processing of dynamic auditory events across both the speech and non-speech domains. Perceptual discrimination improved along with transient lengthening. Accordingly, early MMF was, as a rule, enlarged in case of longer as compared to shorter transients. The 36-ms transitions yielded attention and voicing-dependent deviations from the linear regression of MMF strength on transition duration. Considering the predominance of 30- to 40-ms formant transients across the world's languages, these findings indicate an adaptation or predisposition of the human perceptual system to the spectral/temporal characteristics of prototypical speech sounds. Signal voicing had no significant main effect on MMF strength despite superior perceptual performance in case of voiced as compared to voiceless target stimuli. PMID- 12763200 TI - Posture and mental task performance when viewing a moving visual field. AB - We investigated the characteristics of standing posture and performance of concurrent cognitive tasks in subjects confronted by whole field visual motion. Movements of the head and centre of pressure (COP) were recorded in 12 subjects who performed modified Brooks spatial and verbal tasks when in quiet stance viewing a chequerboard pattern, planar, visual field, moving with uniform velocity (25 degrees /s, 50 degrees /s and 76 degrees /s). Eight subjects were also tested seated to control for the effect of stance. Task load was monitored by heart rate and eye movements were recorded to ensure viewing compliance. Subjects rated their quotidian susceptibility to visual disorientation on a validated scale. In both lateral and antero-posterior directions there were small amplitude but significant increases in COP sway path length and standard deviations of both COP and head sway during exposure to visual motion in proportion to visual flow speed. Performing cognitive tasks during visual motion attenuated sway S.D. The effects on sway of task and visual flow were independent. Visual motion induced a slight tilt and turn of the head and body in the direction of flow together with slight neck flexion. Errors on both verbal and spatial tasks increased >250% during visual motion both when standing and when seated. Ratings of subjects' susceptibility to disorientation were un related to either verbal or spatial task error rates. A current hypothesis is that the enhancement of sway by visual motion is destabilisation. We propose an alternative explanation that sway enhancement could be exploratory 'testing of the ground' movements to check for self motion. Hence decrease in sway magnitude during a cognitive task could be caused by a reduction in exploratory movement because attention is diverted from postural control to a secondary task. Mere passive viewing of a moving visual field may interfere with cognitive tasks possibly because the threat of disorientation by whole field motion diverts attentional resources. PMID- 12763201 TI - Auditory capture of vision: examining temporal ventriloquism. AB - Four experiments investigated whether irrelevant sounds can influence the perception of lights in a visual temporal order judgment task, where participants judged which of two lights appeared first. In Experiment 1, presenting a sound before the first light and after the second light improved performance relative to baseline (sounds appearing simultaneously with the lights), as if the sounds pulled the perception of lights further apart in time. Experiment 2 ruled out an alerting explanation for this effect and indicated that the performance improvement resulted from the second sound trailing the second light. Experiment 3 excluded the possibility that leading or simultaneous sounds were interfering with performance and revealed that only the second sound had an effect within the temporal window known to support multisensory integration. Experiment 4 demonstrated that sounds intervening between the two lights led to a decline in performance, as if the sounds pulled the lights closer together. The results suggest a 'temporal ventriloquism' phenomenon analogous to spatial ventriloquism. PMID- 12763202 TI - Effects of sleep onset on the mismatch negativity (MMN) to frequency deviants using a rapid rate of presentation. AB - This study examined the effects of sleep onset-the transition from a waking, conscious state to one of sleep and unconsciousness-on the mismatch negativity (MMN) following frequency deviants when a rapid rate of stimulus presentation is employed. The MMN is thought to reflect a brief-lasting sensory memory. Rapid rates of stimulus presentation should guard the sensory memory from fading. A 1,000 Hz standard stimulus was presented every 150 ms. At random, on 6.6% of the trials, the standard was changed to either a large 2,000 or a small 1,100 Hz deviant. During alert wakefulness (when subject ignored the stimuli and read a book), the large deviant elicited a larger deviant related negativity (DRN) than did the small deviant. This negativity may be a composite of both N1 and MMN activity while that following the small deviant is probably a 'true' MMN. The large deviant continued to elicit a DRN in relaxed wakefulness (eyes closed) and Stages 1 and 2 of sleep, although it was much reduced in amplitude. A significant MMN was recorded for the small deviant only in alert wakefulness. The failure to observe an MMN to small deviance and the attenuation of the DRN to large deviance at sleep onset therefore is probably not due to a decay of sensory memory. It is more likely that cortical encoding of both the standard and deviant is weakened during sleep onset because of prior thalamic inhibition of sensory input. PMID- 12763203 TI - Event-related potential correlates of the attentional blink phenomenon. AB - The attentional blink phenomenon results from a transitory impairment of attention that can occur during rapid serial stimulus presentation. A previous study on the physiological correlates of the attentional blink employing event related potentials (ERPs) suggested that the P3 ERP component for target items presented during this impairment is completely suppressed. This has been taken to indicate that the target-related information does not reach working memory. To reevaluate this hypothesis, we compared ERPs evoked by detected and missed targets in the attentional blink paradigm. Eighteen subjects performed a rapid serial visual presentation (RSVP) task in which either one target (control condition) or two targets had to be detected. ERPs elicited by the second target were analyzed separately for trials in which the target had been detected and missed, respectively. As predicted, detected targets did elicit a P3 during and after the attentional blink period. No clear P3 was found for detected targets presented before the attentional blink, that is, at lag 1. In contrast, missed targets generally did not evoke a P3. Our results provide evidence that targets presented during the attentional blink period can reach working memory. Thus, these findings contribute to evaluating theories of the attentional blink phenomenon. PMID- 12763204 TI - Electrocortical and behavioral effects of chronic immobility on word processing. AB - Neuroelectrical and subjective effects of chronic motor deficits (severe immobility) on word processing were explored. A total of ten artificially ventilated patients with end-stage amyotrophic lateral sclerosis (ALS), seven patients with tetraplegia due to a high spinal cord injury (SCI), and nine matched controls were examined. Both ALS and SCI patients were almost completely paralyzed; eye, head, and face movements were intact in SCI but not in ALS patients. Event-related brain potentials (ERPs) were recorded during presentation of verbs with strong motor-action associations and nouns with strong visual associations. Subjects estimated the subjective strength of motor and visual associations for each word using a scale from 0 to 5. No between-group difference was found for subjectively rated motor associations. All patients reported much stronger visual associations for both nouns and verbs than healthy subjects. ERPs to both word classes revealed a left occipital negativity in SCI patients at a latency of 200-300 ms, and a marked decrease of the slow positive wave in the interval of 300-800 ms in the two patient groups. Both psychophysical and electrophysiological data suggest compensatory activation of visual information processing areas in severely paralyzed patients. The more pronounced effects in the SCI group as compared with the ALS group may be attributed to somatosensory deafferentation in SCI patients, while the sensory inflow in ALS remains relatively intact. PMID- 12763205 TI - Physical activity in long-term survivors of germ-cell cancer. PMID- 12763206 TI - Levels of absolute survival benefit for systemic therapies of advanced cancer. a call for standards. AB - Research on systemic interventions in patients with advanced stage malignancies should be systematised with an emphasis on the absolute gain in survival for the median patient. Such information is most meaningful with relatively large-scale evidence. Here, we summarise the survival impact of 36 interventions compared against other interventions or no treatment for advanced stage malignancies in meta-analyses of individual patient data or in selected recent (2000-2002) randomised trials with >300 randomised subjects. Although 16 interventions showed a formally statistically significant survival benefit against the comparator arm, this exceeded 3 months in only 7 cases. We propose a standardised categorisation of the median survival prolongation in trials and meta-analyses. Level 0: no proven survival benefit; level I: 0-3 months; level II: >3-6 months; level III: >6-24 months; and level IV: more than 24 months. These standardised levels may be incorporated into clinical practice guidelines for individual care and policy making. PMID- 12763207 TI - Genomics and proteomics in cancer. AB - Cancer development is driven by the accumulation of DNA changes in the approximately 40000 chromosomal genes. In solid tumours, chromosomal numerical/structural aberrations are common. DNA repair defects may lead to genome-wide genetic instability, which can drive further cancer progression. The genes code the actual players in the cellular processes, the 100000-10 million proteins, which in (pre)malignant cells can also be altered in a variety of ways. Over the past decade, our knowledge of the human genome and Genomics (the study of the human genome) in (pre)malignancies has increased enormously and Proteomics (the analysis of the protein complement of the genome) has taken off as well. Both will play an increasingly important role. In this article, a short description of the essential molecular biological cell processes is given. Important genomic and proteomic research methods are described and illustrated. Applications are still limited, but the evidence so far is exciting. Will genomics replace classical diagnostic or prognostic procedures? In breast cancers, the gene expression array is stronger than classical criteria, but in endometrial hyperplasia, quantitative morphological features are more cost effective than genetic testing. It is still too early to make strong statements, the more so because it is expected that genomics and proteomics will expand rapidly. However, it is likely that they will take a central place in the understanding, diagnosis, monitoring and treatment of (pre)cancers of many different sites. PMID- 12763208 TI - The level of physical activity in long-term survivors of testicular cancer. AB - The aim of this study was to estimate the level of physical activity (LPA) in a large cohort of testicular cancer survivors (TCSs) and compare these results with observations from men in the same age range in the general population (GenPop). We also wanted to identify parameters that influenced physical activity. The study populations consisted of 1276 TCSs treated with surgery, radiotherapy or chemotherapy with or without surgery (mean observation time was 12 years), and 20391 male inhabitants from a Norwegian county (GenPop). All completed a question investigating two sub-levels of physical activity. The logistic regression analysis adjusting for different covariates, showed significantly more physically active men among the TCSs compared with the GenPop (43 versus 37%) (adjusted odds ratio (aOR)=1.32 (95% Confidence Interval (CI) 1.10-1.58)). The results indicate that the experience of testicular cancer increases rather than reduces the LPA in TCSs, independent of treatment given. PMID- 12763209 TI - Concurrent hypermethylation of gene promoters is associated with a MSI-H phenotype and diploidy in gastric carcinomas. AB - Changes in the pattern of DNA methylation are among the most common alterations observed in human cancers, such as gastric carcinomas. We analysed in a series of 51 sporadic gastric carcinomas the methylation status of the promoter regions of the hMLH1, CDH1, MGMT and COX2 genes. We aimed to determine the frequency of CpG island hypermethylation and to find out whether the occurrence of concurrent hypermethylation is related to the clinicopathological features of the gastric carcinomas. Using methylation-sensitive restriction analysis/polymerase chain reaction (PCR) and methylation-specific PCR (MSP) strategies, we searched for the presence of hypermethylation on the promoter region of the 4 selected genes. All showed hypermethylation of their promoter regions with frequencies of 37, 51, 61 and 29% for hMLH1, CDH1, MGMT and COX2, respectively. Concurrent hypermethylation was more frequently observed in MSI-H (P=0.0005) and diploid (P=0.029) tumours. Hypermethylation of hMLH1 was associated with MSI-H tumours (P=0.0001), whereas hypermethylation of MGMT was associated with MSI-H (p=0.021) and diploid tumours (p=0.012). Our results indicate that concurrent hypermethylation is a common event in gastric cancer, suggesting that global methylation changes play an important role in the development of sporadic gastric carcinoma. Moreover, inactivation of different gene promoters by hypermethylation is significantly associated with microsatellite instability (MSI-H) and diploidy: hMLH1 determines MSI-H and MGMT the diploid status of gastric carcinomas. PMID- 12763211 TI - Intrinsic variability in the detection of micrometastases in lymph nodes for re staging of colorectal cancer. effect of individual markers and tissue samples. AB - In this study, we investigated whether (a) carcinoembryonic antigen (CEA), cytokeratin-20 (CK-20) and guanylyl cyclase C (GCC) are clinically useful markers for the molecular detection of submicroscopic metastases in colorectal cancer (CRC) and (b) whether overexpression of CEA, CK-20 and GCC can be reliably detected in formalin-fixed, paraffin-embedded tissues as well as frozen lymph nodes. We studied 175 frozen lymph nodes and 158 formalin-fixed, paraffin embedded lymph nodes from 28 cases of CRC. CEA or CK-20 or GCC-specific polymerase chain reaction (PCR) was carried out on mRNA transcripts extracted from the nodal tissues. Ten out of 11 Dukes' B CRC cases had detectable CEA and CK-20 while 6 out of 11 Dukes' B CRC cases had detectable GCC. In general, the difference of re-staged cases when comparing frozen and paraffin-embedded samples was marked; the only statistically significant correlation between frozen and paraffin tissue was for the CEA marker. Our results indicated a high incidence (>50%) of detecting micrometastases in histologically-negative lymph nodes at the molecular level. PMID- 12763210 TI - Fulvestrant (Faslodex) versus anastrozole for the second-line treatment of advanced breast cancer in subgroups of postmenopausal women with visceral and non visceral metastases: combined results from two multicentre trials. AB - The efficacy of fulvestrant (Faslodex), a novel oestrogen receptor (ER) antagonist that downregulates the ER and has no known agonist effects, was compared with the aromatase inhibitor anastrozole (Arimidex) for the second-line treatment of advanced breast cancer in postmenopausal women with visceral and non visceral metastases. Assessment was by means of a retrospective subgroup analysis of combined data from two randomised, phase III trials. Objective response (OR) rates were similar in patients treated with fulvestrant and anastrozole, respectively (21.9% versus 19.3%-patients with no visceral metastases; 15.7% versus 13.2%-all of the patients with visceral metastases; 18.8% versus 14.0% patients with visceral metastases only). The proportion of patients with clinical benefit (CB) was also similar between treatments and between subgroups with and without visceral disease. Fulvestrant is at least as effective as anastrozole, providing a valuable treatment option for advanced breast cancer in postmenopausal women with visceral metastases who have failed on prior endocrine therapy. PMID- 12763212 TI - Prognostic significance of K-ras, p53, bcl-2, PCNA, CD34 in radically resected non-small cell lung cancers. AB - The aim of this study was to investigate the prognostic significance of a panel of biological parameters in patients with radically resected non-small cell lung cancers (NSCLC). 269 cases with pathological stage I-IIIA NSCLC were retrospectively analysed. Immunohistochemistry was performed to detect protein expression of p53, bcl-2, proliferating cell nuclear antigen (PCNA) and CD34. Polymerase chain reaction (PCR)/direct nucleotide sequencing method was used to detect mutations in K-ras (codons 12, 13, 61, exons 1-2). The Kaplan-Meier estimates of survival were calculated for clinical and biological variables using the Cox model for multivariate analysis. Histological subtype and the pathologic tumour extension (pT) were the most powerful clinical-pathological prognostic factors for survival (P=0.030 and P=0.031, respectively), whereas among the biological parameters, p53 overexpression (P=0.032) and K-ras mutation (P=0.078) had a negative prognostic role, as demonstrated by multivariate analysis. Conversely, bcl-2, PCNA and CD34 expression were not correlated with survival. Statistically significant associations between p53 expression and the squamous cell carcinoma (SCC) subtype, bcl-2 expression and SCC subtype, K-ras mutation and p53 negative expression, p53 and bcl-2, bcl-2 and PCNA overexpression were observed. In conclusion, some biological characteristics such as the K-ras and p53 status may provide useful prognostic information in resected NSCLC patients, in addition to the classical clinico-pathological parameters. However, further studies are needed to clarify the value of adopting biological prognostic factor into clinical practice. PMID- 12763213 TI - Oestrogen receptor beta (ERbeta) is abundantly expressed in normal colonic mucosa, but declines in colon adenocarcinoma paralleling the tumour's dedifferentiation. AB - Oestrogen Receptor beta (ERbeta) may protect against prostate and mammary cell proliferation and malignant transformation. Epidemiological studies indicate that oestrogens may reduce colon cancer risk. Since ERalpha is minimally expressed in normal and malignant colon, the aim of this study was to investigate the expression of ERbeta in both normal colonic wall and colon cancer. ERbeta expression was evaluated by immunohistochemistry in 90 cases of colon adenocarcinoma and nearby (>30-cm away) normal colonic wall, using a monoclonal antibody. Moderate or strong nuclear immunostaining was detected in superficial and crypt epithelium, endothelial cells, vascular smooth muscle cells, lymphocytes, enteric neurons and smooth muscular cells of the normal colonic wall. Superficial epithelial cells in normal colon demonstrated a significantly higher ERbeta expression than colon adenocarcinoma cells in both genders. The decline in ERbeta expression paralleled the loss of differentiation of malignant colon cells, regardless of the tumour's localisation. These findings suggest a protective role for ERbeta against colon carcinogenesis. PMID- 12763214 TI - Development of an EORTC disease-specific quality of life questionnaire for use in patients with liver metastases from colorectal cancer. AB - Quality of life (QL) is an important outcome measure within clinical trials. This paper describes the development of a QL module for patients with liver metastases from colorectal cancer (CRC) to supplement the European Organization for Research and Treatment of Cancer (EORTC) core QL questionnaire, the EORTC QLQ-C30. Phases 1-3 of the EORTC QL Group guidelines for developing QL modules were followed. The literature search generated 71 QL issues. Semi-structured interviews with eight healthcare professionals and 47 patients from the United Kingdom, France, Germany and Austria reduced the list to 23 issues. Questionnaire items were formulated to be compatible with the EORTC QLQ-C30. The provisional module was further tested in 102 patients resulting in a 21-item module, the QLQ-LMC21 (Liver Metastases Colorectal). A combination of the core questionnaire and the QLQ-LMC21 will provide essential QL information regarding the use of treatments in both the curative and palliative settings. PMID- 12763215 TI - Phase II trial with S-1 in chemotherapy-naive patients with gastric cancer. A trial performed by the EORTC Early Clinical Studies Group (ECSG). AB - S-1 is a new oral fluorinated pyrimidine derivate, in which the oral 5 fluorouracil (5-FU) prodrug, tegafur, was combined with two 5-FU-modulating substances, 5-chloro-2,4-dihydroxypyridine (gimeracil), and potassium oxonate (oteracil), at a molar ratio of 1:0.4:1. The final mechanism of action is exerted by 5-FU. The present study is the first European phase II trial of S-1 in gastric cancer. The primary study objectives were the safety, toxicity and activity of S 1 in non-pretreated patients with gastric cancer. The secondary objective was the duration of response. Patients had to have histologically- or cytologically verified metastatic or locally advanced, unresectable gastric cancer; S-1 was administered orally twice daily at 40, then 35 mg/m(2) for 28 days every 5 weeks. The starting dose of 40 mg/m(2) was found to be intolerable due to significant non-haematological toxicity, and this dose was rapidly reduced to 35 mg/m(2) twice daily. Of the 7 patients enrolled at the 40 mg/m(2) level, only 3 were evaluable. At 35 mg/m(2), a response rate of 26.1% (95% Confidence Interval (CI) 12.0-45.1%) in 23 enrolled patients, and 31.6% (C.I. 14.7-53.0%) in 19 evaluable patients according to an independent radiology review, was found. The median duration of response at 35 mg/m(2) (6 patients) was 223 days (range, 108-828 days), and of stable disease was 111 days (range 68-411 days). S-1 can be administered with an acceptable safety and toxicity in European patients at a dose of 35 mg/m(2) days 1 - 28 every 5 weeks and is associated with a moderate response rate similar to the results achieved with other fluoropyrimidines. PMID- 12763216 TI - Temozolomide in patients with advanced non-small cell lung cancer with and without brain metastases. a phase II study of the EORTC Lung Cancer Group (08965). AB - This study was performed to evaluate the activity of single-agent temozolomide in two groups of chemotherapy-naive non-small cell lung cancer (NSCLC) patients, with (12 patients) and without (13 patients) brain metastases (BM). Patients in both groups were treated with temozolomide 200 mg/m(2)/day, administered orally for 5 consecutive days of a 28-day cycle. Treatment was continued for up to six cycles, disease progression or unacceptable toxicity. The median number of received cycles was only one in the group with and two in the group without BM, and early disease progression was the main reason for treatment discontinuation. Toxicity was moderate-in the group of patients with BM, the most frequently observed grade 3 or 4 side-effects included thrombocytopenia (17%), granulocytopenia (17%), lethargy (17%); other neurological (17%) and other genitourinary toxicity (17%). Patients without BM experienced anaemia (15%), thrombocytopenia (23%), nausea (15%) and lethargy (15%). This trial was designed according to Simon one-sample two-stage testing procedure and both groups of patients were assessed separately. No objective response was observed in either group and the study was closed after the first step of accrual with the conclusion of a lack of therapeutic activity of single-agent temozolomide in patients with stage IV NSCLC. PMID- 12763217 TI - An EORTC phase I study of capecitabine (Xeloda) in combination with fixed doses of cyclophosphamide and epirubicin (cex) as primary treatment for large operable or locally advanced/inflammatory breast cancer. AB - In breast cancer, chemotherapy regimens that include infusional 5-fluorouracil (5 FU) lead to high response rates, but require central venous access and pumps. To avoid these inconveniences, we substituted infusional 5-FU with capecitabine. The main objective of this study was to determine the maximum tolerated dose (MTD) of capecitabine when given in combination with fixed doses of epirubicin and cyclophosphamide (100 and 600 mg/m(2) day 1 every (q) 3 weeks) as primary treatment for large operable or locally advanced/inflammatory breast cancer without distant metastasis. Capecitabine was escalated from 750 mg/m(2) twice a day (bid) to 1250 mg/m(2) bid from day 1 to day 14 in four dose levels. Dose escalation was permitted if 0/3 or 1/6 patients experienced dose-limiting toxicity (DLT). A total of 23 patients were included and 117 courses were administered. At dose level 4, 2 of 2 patients presented DLTs defining the MTD. A high rate of capecitabine treatment modification was required with capecitabine 1050 mg/m(2) bid (dose level 3). 19 patients achieved an objective response (83%). In conclusion, we believe that capecitabine 900 mg/m(2) bid (dose level 2) is the recommended dose in combination with epirubicin 100 mg/m(2) and cyclophosphamide 600 mg/m(2). The acceptable toxicity profile and encouraging activity of this regimen warrant further evaluation. PMID- 12763218 TI - Viridans streptococci bacteraemia in children with fever and neutropenia: a case control study of predisposing factors. AB - Viridans streptococci (VS) are an increasing cause of bacteraemia in neutropenic patients with cancer. Case-control studies of predisposing factors for acquisition of this infection in children are not published. Between January 1989 and December 1999, 168 episodes of bacteraemia in 161 children with fever and neutropenia of haemato-oncology origin were analysed. 15 cases (9%) in 15 patients were caused by VS. Each case patient was compared with 6 matched control patients; 2 with other Gram-positive cocci (group 2), 2 with gram-negative bacilli bacteraemia (group 3) and two children with fever and neutropenia without bacteraemia (group 4). The median age of patients was 4.1 years (range: 2-15 years). 87% of children had acute leukaemia or lymphomas. Pneumonia was the predominant clinical focus (70%). Shock was observed in 13% of patients. ARDS was observed in one child who died of this complication. Multivariate analysis of risk factors for the development of VS bacteraemia showed that two factors were independent predictors: high doses of cytosine-arabinoside (ARA-C) as part of the chemotherapy treatment (Odds Ratio (OR): 9.3; Confidence Interval (CI) 1.56-55.5) (P<0.014) and the presence of pneumonia (OR: 1.36: CI 2.27-81.9) (P<0.0043). We propose that further studies are warranted to confirm these results. PMID- 12763219 TI - Analysis of survival of mesothelioma cases in the Italian register (ReNaM). AB - The Italian National Mesothelioma Register (ReNaM) was set up at ISPESL (the National Institute for Occupational Safety and Prevention) in 1993. Five Italian regions (Piedmont, Liguria, Emilia-Romagna, Tuscany and Puglia, with a total of approximately 17500000 inhabitants) agreed to record mesothelioma cases according to guidelines established by ISPESL, to define exposure to asbestos and transmit the data to ISPESL. We describe an analysis of survival of 429 mesothelioma cases 392 pleural, 34 peritoneal and 3 in the pericardium-diagnosed during 1997, with variable follow-up from June 1999 to December 2001. The Kaplan-Meier method was used to estimate survival rates, the log rank non-parametric test and Cox proportional hazard model to assess the role of prognostic factors such as age, gender, morphology, level of diagnostic certainty and modality of exposure. Median survival was 275 days (95% confidence interval (CI) 241-309) for pleural mesotheliomas and 157 days (95% CI: 118-196) for peritoneal mesotheliomas. Survival after diagnosis of malignant pleural mesothelioma showed a statistically significant linear trend for age group at diagnosis, for males and females (P=0.006 and 0.008, respectively). The Cox proportional hazard model gave an adjusted relative risk (RR(adj)), for the fibrous histotype, of 2.96 (95% CI: 1.28-6.81; P=0.012) compared with cases with unspecified morphology; for epithelioid and biphasic morphologies, the risk was lower than unity. There was no significant difference in survival for cases with confirmed exposure (occupational, household or environmental) or without. PMID- 12763220 TI - Differential expression of inducible nitric oxide synthase and peroxisome proliferator-activated receptor gamma in non-small cell lung carcinoma. AB - Both inducible nitric oxide synthase (iNOS) and peroxisome proliferator-activated receptor gamma (PPARgamma) are closely associated with the development of human cancer. Although the expression of iNOS has been studied in non-small cell lung carcinoma (NSCLC), the level of PPARgamma has not been examined in tumorous and non-tumorous tissues from NSCLC. The present study analysed the levels of both iNOS and PPARgamma in NSCLC tissues and in lung cell lines. The possible role of these two molecules in the carcinogenesis of lung cancer was investigated. The expression of iNOS was significantly higher in the tumorous tissues than in the non-tumorous ones. In contrast to this pattern of iNOS protein expression, the level of PPARgamma was much lower in the tumorous tissues than in the non tumorous samples. A similar result was also obtained in vitro using human lung cancer cell lines and normal lung cells. Immunohistochemical examination revealed that PPARgamma expression in the non-tumorous tissues was more likely to be located in the nucleus whereas it was present in both the nucleus and cytoplasm of the tumorous tissues. The intensity of iNOS expression was stronger in the nucleus than in the cytoplasm of the tumorous tissues. More than 50% of the cases tested did not express iNOS protein in the non-tumorous tissues. Statistical analysis indicated a negative correlation between iNOS and PPARgamma levels in the NSCLC tissues. In conclusion, this study demonstrated differing expressions for iNOS and PPARgamma in NSCLC tissues. Since activated PPARgamma is able to inhibit the expression of iNOS and the generation of iNOS is particularly associated with the inflammatory and environmental factors of lung cancer risk, this discrepant expression pattern may be associated with the pathogenesis of NSCLC. PMID- 12763221 TI - The activation of an extracellular signal-regulated kinase by oestradiol interferes with the effects of trastuzumab on HER2 signalling in endometrial adenocarcinoma cell lines. AB - Cellular response to oestradiol stimuli is mediated both by oestrogen receptor (ER) binding to oestrogen response elements (EREs) and by non-nuclear actions like activation of mitogen-activated protein kinase (MAPK) signal transduction. Therefore, oestradiol stimuli might be able to interfere with the action of antitumoral substances directed against receptor tyrosine kinase signalling. We investigated the effect of oestradiol on the inhibition of HER2 signalling by trastuzumab (Herceptin) in two human endometrial adenocarcinoma cell lines. Activation of the extracellular signal-regulated kinase (ERK-1/2), a major mediator of HER2 signalling, was measured by means of western blotting experiments and ERE activation was determined in transient reporter-gene assays. In endometrial Ishikawa and HEC-1A adenocarcinoma cells, HER2 signalling was inhibited by trastuzumab only in the absence of oestradiol. We were able to demonstrate that oestradiol counteracted the inhibitory effects of trastuzumab by rapid phosphorylation of ERK-1/2, a kinase downstream of the HER2 receptor. The pure anti-oestrogen ICI 182,780 was able to restore both the trastuzumab triggered inhibition of the ERK-1/2 pathway and the antiproliferative action of this substance in Ishikawa cells. Our data suggest that combinations of trastuzumab with anti-oestrogens may be effective in the treatment of endometrial cancers with a positive ER and HER2 receptor status. PMID- 12763222 TI - Thymidylate synthase inhibition triggers apoptosis via caspases-8 and -9 in both wild-type and mutant p53 colon cancer cell lines. AB - Thymidylate synthase (TS) is an important target for chemotherapy and increased levels are associated with resistance to colorectal cancer chemotherapy. TS can be inhibited by 5-fluorouracil (5-FU) and antifolates, ultimately resulting in apoptosis. We aimed to clarify whether activation of caspases and Fas signalling are crucial for the onset of apoptosis after specific inhibition of TS and whether p53 plays a role in activation of these downstream processes. For this purpose, wild-type (wt) and mutant (mt) p53 colon cancer cell lines, Lovo and WiDr, respectively, transfected with mt- and wt-p53, were treated with the specific TS inhibitor, AG337. Treatment with 10xIC(50) values of AG337 for 48 h resulted in S phase arrest in all Lovo and WiDr cells (up to 50% of cells being in S phase), irrespective of their p53 status. After 72 h, the induction of apoptosis was most pronounced in the AG337-sensitive cells. Approximately 30% apoptosis was detected in all of the WiDr cells, 20% in Lovo li (non-functional p53), 12-14% in Lovo 92 and B2 (wt p53) and only 7% in Lovo 175x2 cells (mt p53 transfected). The induction of apoptosis in Lovo cells, as determined using the classical sub-G1 peak after propidium iodide (PI) staining, was associated with an increase in the expression of Fas receptor. In addition, synergistic increases in apoptosis from approximately 10 to 35% after 48 h could be detected after simultaneous treatment of AG337 and the Fas activator antibody, CH11. Only additive effects were measurable in WiDr cells, without an increase in Fas receptor expression. Surprisingly, the Fas inhibitor, ZB4, could not decrease the amount of cell death in both cell lines after AG337 treatment. In contrast, simultaneous exposure of Lovo and WiDr cells to AG337 and inhibitors of caspases 8, 9 and 3 caused a decrease in the number of apoptotic cells compared with AG337 exposure alone. Inhibition of apoptosis by approximately 10-80% in Lovo and approximately 70-80% in WiDr cells could be detected. In conclusion, these results indicate that apoptosis induced after specific inhibition of TS is mediated via the caspases, but without clear involvement of Fas signalling. The status of p53 did not affect the onset of apoptosis by these caspases. PMID- 12763223 TI - Trail-induced apoptosis and interaction with cytotoxic agents in soft tissue sarcoma cell lines. AB - Five human soft tissue sarcoma (STS) cell lines (HTB-82 rhabdomyosarcoma, HTB-91 fibrosarcoma, HTB-92 liposarcoma, HTB-93 synovial sarcoma and HTB-94 chondrosarcoma) were analysed for their sensitivity to tumour necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) and the function of the TRAIL apoptotic pathway in these cells. TRAIL induced significant apoptosis (>90%) in HTB-92 and HTB-93 cells, whereas no effect was observed in HTB-82, HTB-91 and HTB 94 cells. TRAIL-Receptor 1 (TRAIL-R1) was expressed in TRAIL-sensitive HTB-92 and HTB-93 cell lines, but not in TRAIL-resistant HTB-91 and HTB-94 cells. HTB-82 cells, which expressed the long (c-FLIP(L)) and short (c-FLIP(S)) splice variants of the FLICE-like inhibitory protein (FLIP), were resistant to TRAIL in spite of the presence of TRAIL-R1. TRAIL-R2,-R3,-R4 and osteoprotegerin (OPG) expression did not correlate with TRAIL sensitivity. Coincubation of TRAIL and doxorubicin led to the overexpression of TRAIL-R2 resulting in a synergistic effect of doxorubicin and TRAIL in TRAIL-sensitive cell lines and in the overcoming of TRAIL-resistance in all of the TRAIL-resistant cell lines, except HTB-91, which lacked caspase 8 expression. These data suggest that TRAIL, either as a single agent or in combination with cytotoxic agents, might represent a new treatment option for advanced STS, which constitutes a largely chemotherapy-resistant disease. PMID- 12763224 TI - Inexpensive ethography using digital video. AB - We describe an inexpensive method for digital video recording of behavioral experiments and present a simple, freely-redistributable software tool enabling ethographic analysis of these recordings via classification of video frames into user-defined categories. While high-end commercial solutions already exist for these purposes, we focus on minimizing equipment expenses and complexity for research projects or undergraduate laboratory courses employing compatible behavioral paradigms. PMID- 12763225 TI - Fluorescence digital photomacrography. AB - Low-power photographs of brain sections have been needed for neuroanatomical studies. This paper describes a setup for fluorescence digital photomacrography. High-resolution digital images were obtained with commercially available digital cameras combined with an oblique fluorescence illumination and filters for excitation and emission. The image of an entire coronal section of the macaque brain could be taken with a single exposure. The present photographic system would be useful not only for neuroanatomical, but also for histological, embryological or pathological studies that require low-magnification fluorescence images. PMID- 12763226 TI - A cuvette-based fluorometric analysis of mitochondrial membrane potential measured in cultured astrocyte monolayers. AB - Mitochondrial membrane potential (Deltapsi(M)) plays a key role in coordinating mitochondrial function and cell biology in general. In astrocytes, Deltapsi(M) is an important indicator of the health of these brain cells and their response to traumatic and hypoxic injury. We have shown previously how fluorescent signals can be measured from cells attached to a coverslip in a standard cuvette with a fluorometer and modulated using a cuvette perfusion system (Pflugers Arch-Eur. J. Physiol. 421 (1992) 400). Here we report on how this method can be employed to characterize the actions of a number of potentiometric fluorescent cationic dyes, including JC-1, Rh123 and TMRM, for their ability to monitor Deltapsi(M) in primary cultures of intact astrocytes. All dyes detected the reversible depolarization produced by brief exposure to the mitochondrial uncoupler protonophore FCCP, which short circuits and dissipates Deltapsi(M). Qualitatively similar responses were measured after treatment with either azide, an inhibitor of complex IV in the mitochondrial respiratory chain, or the oxidant H(2)O(2). Cell depolarization with high potassium modified the responses to FCCP. The time courses of these responses differed in a manner dependent on the particular dye used and in a way that correlated with expected permeation rate. The merits and pitfalls of these different potentiometric dyes for monitoring Deltapsi(M) are discussed. PMID- 12763227 TI - A protocol for isolation and biochemical characterization of stigmoid bodies from rat brain. AB - Stigmoid bodies (SBs) are structures present in the cytoplasm of neurons. Many brain regions including hypothalamus, thalamus, amygdala, septum, hippocampus, colliculi, and brainstem contain neurons with at least one SB. Despite this widespread distribution their function remains unknown. SBs contain a brain protein called huntingtin-associated protein 1 (HAP1) and have more recently been found to contain the apolipoprotein E receptor LR11 (Lipoprotein Receptor containing 11 LDL binding domains, also called SorLA for sorting protein-related receptor containing LDLR class A repeats) and sortilin. To provide a first step towards further identification of their components and perhaps shed some light on their neurobiological role, we have developed a method for isolating SBs from rat brain. The protocol relies on a combination of centrifugational forces, sucrose gradient, and immunoisolation. Samples enriched in SBs were incubated with antibodies to HAP1B or to LR11 followed by incubation with FITC conjugated secondary antibodies. Anti-FITC coated beads were incubated with samples and SB bead complexes formed were separated by magnetic sorting without pelleting the complexes during the isolation procedure. Immunopurified SBs, visualized by light and electron microscopy, show similar ultrastructure to those present in neurons. PMID- 12763228 TI - A computerized image analysis system for quantitative analysis of cells in histological brain sections. AB - We propose a reliable method for automatic counting of cells in brain sections labeled with different antibodies (against NeuN, parvalbumin, GABA and c-Fos) and in Nissl-staining. Images of stained sections are converted to binary images by thresholding. Clusters of 'ON pixels' (value of 1) corresponding to cell bodies are selected based on size. The parameters of the algorithm (intensity range and cluster-size) are adjusted for different methods of staining according to expert knowledge. The automatic cell counting method (ACCM) provides correct counting results, as demonstrated by a comparison of computational results with counts gained by human experimenters and with a commercially available image analysis system. On the basis of ACCM counts, small and perhaps physiologically relevant differences in the number of labeled cells can be revealed, as demonstrated here for the GABAergic system following electrical stimulation. PMID- 12763229 TI - Biolistic transfection of organotypic cultures of rat visual cortex using a handheld device. AB - Our aim is the biolistic transfection of organotypic cultures of rat visual cortex with plasmids coding for neurotrophic factors, which then become expressed for limited periods of time during postnatal ontogenesis. Out of two commercially available devices, we adopted the handheld 'Helios Gene Gun' instead of the stationary PDS-1000He (both Biorad, Munich, Germany). This device allows multiple transfections of single targets and the transfection of a distinct part of a co culture when utilising an aperture. Unfortunately, the most detailed protocols are limited to the stationary device and not compatible with the hand-held device. We report here the construction of a support for the gene gun including an aperture and the establishment of a protocol to efficiently transfect rat cortical slice cultures. We achieve a high degree of co-expression of independent plasmids coated on the same particles. The expression of the neurotrophin plasmids is demonstrated on mRNA and protein level. PMID- 12763230 TI - Quantification of antioxidant activity in brain tissue homogenates using the 'total equivalent antioxidant capacity'. AB - We demonstrate herein that the standard 'Trolox equivalent antioxidant capacity' (TEAC) assay, typically utilized to quantify total antioxidant levels within plasma, can also be utilized for tissue homogenates. Normal mice and transgenic mice lacking apolipoprotein E were subjected to a diet including iron as a generic pro-oxidant for 1 month (which has been shown to induce oxidative damage in our prior studies) and homogenates of brain tissue were subjected to the TEAC assay. Levels of the endogenous antioxidant glutathione levels were also monitored by HPLC. As described previously, ApoE-deficient mice expressed increased levels of glutathione; total antioxidant levels, as determined by TEAC, were increased to a similar extent. The increase in total antioxidant levels, as determined by TEAC, following dietary iron challenge paralleled the increase in glutathione levels, as determined by HPLC. These findings indicate that the TEAC assay may be useful for tissue homogenates. The rapid nature of this assay compared to HPLC, coupled with its lack of requirement for sophisticated equipment, makes it well suited for analyses of multiple tissue samples. PMID- 12763231 TI - Magnetoneurography: recording biomagnetic fields for quantitative evaluation of isolated rat sciatic nerves. AB - Magnetoneurography (MNG) is a technique to record the biomagnetic action fields of peripheral nerves. The benefits of MNG in contrast to electroneurography include the decreased signal disturbance caused by surrounding biological tissues and the use of a calibration pulse, both of which contribute to high reproducibility. MNG has proven to be a valuable tool to quantitate peripheral nerve regeneration in rabbits. However, the most commonly used model to study the peripheral nervous system is the rat sciatic nerve. Until now, the small size of the nerve impeded accurate MNG measurements in rat. This report describes a custom made recording chamber that allows accurate control of conduction distances and temperature and enables adequate MNG measurements of isolated sciatic nerves of Wistar rats. We applied biphasic stimulation with optimized grounding to reduce the stimulus artefact. A high reproducibility of signals was demonstrated. 'Ex vivo' nerve viability was assured for at least 2 h after dissection. In conclusion, MNG is a powerful tool to quantitatively evaluate the function of rat sciatic nerves and will be used for the early assessment of nerve regeneration. PMID- 12763232 TI - A new extensive approach to single unit responses using multisite recording electrodes: application to the songbird brain. AB - One of the most challenging issues in neuroethology concerns the neural substrates for song production, perception and learning in songbirds. However, electrophysiological studies of the song system of songbirds are often fragmented and centered on a small number of selected neurons. Here, we have developed a new extensive approach to record a great number of single units in the brain of a songbird, the European starling. The aim of this approach is to formulate quantitative assumptions about the electrophysiological characteristics of the brain nucleus investigated: e.g. the proportion of auditory neurons, neuronal selectivity, etc. We applied a mapping method using multisite recording electrodes and online isolation of single-units, without preselecting neurons with a search stimulus. As an example of the application of this technique, we have mapped the responses to a variety of natural and artificial acoustic stimuli recorded systematically throughout the HVC of both awake and anaesthetized male starlings. This method appears to be powerful and allowed us to quantitatively compare responses obtained in awake and anaesthetized birds by recording over 1000 single units. We think that, in future, this will enable us to characterize and compare parts of nuclei or entire nuclei and to better understand how the song system works. PMID- 12763233 TI - Using genetic algorithms to find the most effective stimulus for sensory neurons. AB - Genetic algorithms (GAs) can be used to find maxima in large search spaces in a relatively short period of time. We have used GAs in electrophysiological experiments to find the most effective stimulus (MES) for sensory neurons in the cochlear nucleus and inferior colliculus of anaesthetised guinea pigs. The MES is the stimulus that elicits the greatest number of spikes from a unit. We show that GAs provide an effective means of determining the best combination of up to four parameters for sinusoids with amplitude modulation. Using GAs, we have found tuning to modulation frequencies as a function of carrier frequency, sound level and temporal asymmetry. These results demonstrate the suitability of GAs in electrophysical experiments for estimating the position of the most effective stimulus in a specified parameter space. PMID- 12763234 TI - Labeling Schwann cells with CFSE-an in vitro and in vivo study. AB - Schwann cell (SC) transplantation is a promising strategy for axonal regeneration in the nervous system. Identifying the grafted SCs is an important aspect of this approach. The current study sought to establish a simple, reliable, fluorescent labeling method for SCs with a lipophilic molecule, 5-(and-6)-carboxyfluorescein diacetate succinimidyl ester (CFSE). Human SCs were incubated with varying concentrations of CFSE for different time periods. Based on the viability of labeled SCs and its plating efficiency, 1 min incubation with 5 microM CFSE at 37 degrees C was selected as the optimal labeling condition. Flow cytometric analysis and fluorescence microscopy demonstrated that the fluorescence of labeled SCs would fade over 4 weeks. Immunostaining for the phenotypic expression of SC markers, including S100, GFAP, P75, and MHC-I/II at 1 and 4 weeks after incubation with CFSE showed no difference between labeled and non-labeled SCs. Mixed cultures of labeled human SCs and rat SCs for 48 h were performed in triplicate and demonstrated that no leakage of dye from labeled SCs in cell culture occurred across species. A total of 14 injections of 2x10(5) labeled SCs were performed within the spinal cord at T8 and/or L1 level in 9 nude rats. The animals were euthanized at 1 (6 injections) and 4 weeks (8 injections). Grafted labeled SCs survived for at least 4 weeks, and could be easily recognized in the nude rat spinal cord without leakage of dye to surrounding cells. The SCs migrated in white and gray matter 3-6 mm away from the injection and in the central canal for up to 12 mm. These results suggest that CFSE can be used as a fluorescent tracer of human SCs for both in vitro and in vivo studies, for a period of at least 4 weeks. PMID- 12763235 TI - High-resolution voxelation mapping of human and rodent brain gene expression. AB - Voxelation allows high-throughput acquisition of multiple volumetric images of brain gene expression, similar to those obtained from biomedical imaging systems. To obtain these images, the method employs analysis of spatially registered voxels (cubes). For creation of high-resolution maps using voxelation, relatively small voxel sizes are necessary and instruments will be required for semiautomated harvesting of such voxels. Here, we describe two devices that allow spatially registered harvesting of voxels from the human and rodent brain, giving linear resolutions of 3.3 and 1 mm, respectively. Gene expression patterns obtained using these devices showed good agreement with known expression patterns. The voxelation instruments and their future iterations represent a valuable approach to the genome scale acquisition of gene expression patterns in the human and rodent brain. PMID- 12763236 TI - Electrophysiological, mechanosensitive responses of Xenopus laevis oocytes to direct, isotonic increase in intracellular volume. AB - An intra-oocyte injection method for obtaining the electrophysiological response of follicle-enclosed Xenopus laevis oocytes to an increase in intracellular volume (i.e. stretch) without changing the extracellular medium is described. The response comprised a 'stretch-activated' (SA) current which was evoked by injection of an isotonic 14-70 nl droplet and had a transient, smooth profile. Ionic substitution experiments revealed that the current was carried mainly by Na(+), K(+) and Cl(-) and had a reversal potential of about -2 mV. A similar result was obtained from experiments in which the holding potential was varied between -40 and +10 mV whilst repeatedly inducing the SA current. On average, the channel was blocked 60% by 10 microM gadolinium chloride, 58% by 50 microM amiloride, 11% by 50 microM 4,4'-diisothiocyanatostilbene-2, 2'-disulfonic acid and 63% by 50 microM 4-acetamido-4'-isothiocyanato-stilbene-2-2'-disulfonic acid. Maturation of the oocytes with 100 microM progesterone reduced the mechanosensitivity 12-fold. This injection technique is compared with other methods of eliciting mechanosensitive (MS) currents in X. laevis oocytes. These observed characteristics of the SA current are discussed in relation to the oocytes' endogenous MS cation and anion channels. PMID- 12763237 TI - Efficient gene delivery to primary neuron cultures using a synthetic peptide vector system. AB - A bi-functional, 31 amino acid synthetic peptide (polylysine-molossin) was evaluated for gene delivery to primary cultures of rat cerebral cortex neurons. Polylysine-molossin consists of an amino terminal domain of 16 lysines for electrostatic binding of DNA, and a 15 amino acid, integrin-binding domain at the carboxyl terminal. High levels of gene delivery were obtained with 20-30 microM chloroquine, with a synthetic fusogenic peptide at an optimal DNA:polylysine molossin:fusogenic peptide w/w ratio of 1:3:0.2, and with the addition of low concentrations of Lipofectamine 2000 at an optimal DNA:polylysine molossin:Lipofectamine 2000 w/w ratio of 1:3:0.5. With the best combination, >30% of neurons strongly expressed the beta-galactosidase reporter gene, with no observable toxicity. DNA concentrations >2 microgram/ml were essential for efficient gene delivery. This synthetic peptide provides a safe, readily standardised and flexible DNA vector system well suited to ex vivo gene delivery to neurons for experimental and clinical applications. PMID- 12763238 TI - Using ocular dominance to infer the depth of the visual input layers of V1 in behaving macaque monkey. AB - The goal of this study was to use the ocular dominance properties of multiple unit activity in area V1 of the visual cortex of the behaving rhesus monkey to infer the depth of the visual input layers. Multiple unit activity was examined with a recording electrode at different depths (in 100 micrometer increments) within V1 for responses to a visual stimulus presented to the dominant and non dominant eye. The cortical depth at which there was a maximal difference in unit firing rate between the dominant and non-dominant eye was used to infer the depth of the visual input layers of V1. This depth was found to vary from 0.8 to 1.2 mm below the cortical surface. This range of depths overlaps with the approximate location of lamina IVc, which is the major recipient of visual fibres from the lateral geniculate nucleus. PMID- 12763240 TI - A Cartesian coordinate system for human cerebral cortex. AB - The most commonly used method for specifying the locations of functional areas in the human cerebral cortex is the coordinate system of Talairach and Tournoux (Co planar Stereotaxic Altas of The Human Brain (1988) Georg Thieme Verlag, Stuttgart). It was designed to locate subcortical nuclei by reference to an axis joining the anterior and posterior commissures. The coordinate system has difficulties, however, when applied to cortical locations: (1) it can be difficult to locate the posterior commissure (PC); (2) the fundamental axis is short, and errors in specifying the axis lead to large errors at the cortical surface; (3) there is no normalisation for brain size. We sought to rectify these problems with a new coordinate system, the Sydney system, in which the fundamental axis runs in the medial sagittal plane from the anterior edge of the corpus callosum to the posterior end of the parieto-occipital sulcus. Normalisation is achieved by dividing all distances by the length of the fundamental axis. Using functionally important points and anatomical landmarks on cadaveric specimens and magnetic resonance images (MRI), three-dimensional coordinates were measured in both the Talairach and Sydney systems. The Sydney system has the following advantages over the Talairach system: (1) the fundamental axis is more than four times longer and is easier to identify; (2) the Sydney system is more precise, in that it reduces the spread of points across the sample; (3) the normalised coordinates allow locations to be compared across individuals, regardless of brain size. We conclude that for the mapping of cortical areas, the Sydney system is potentially an improvement on Talairach's. PMID- 12763239 TI - Histological preparation of developing vestibular otoconia for scanning electron microscopy. AB - The unique nature of vestibular otoconia as calcium carbonate biominerals makes them particularly susceptible to chemical deformation during histological processing. We fixed and stored otoconia from all three otolith endorgans of embryonic, hatchling and adult Japanese quail in glutaraldehyde containing either phosphate or non-phosphate buffers for varying lengths of time and processed them for scanning electron microscopy. Otoconia from all age groups and otolith endorgans processed in 0.1 M phosphate buffer (pH 7.4) showed abnormal surface morphology when compared to acetone fixed controls. Otoconia processed in 0.1 M sodium cacodylate or HEPES buffered artificial endolymph (pH 7.4) showed normal morphology that was similar to controls. The degree of otoconial deformation was directly related to the time exposed to phosphate buffer. Short duration exposure produced particulate deformations while longer exposures resulted in fused otoconia that formed solid sheets. Otoconial surface deformation and fusing was independent of the glutaraldehyde component of the histological processing. These findings should help vestibular researchers to develop appropriate histological processing protocols in future studies of otoconia. PMID- 12763241 TI - Models for neuro-oncological preclinical studies: solid orthotopic and heterotopic grafts of human gliomas into nude mice. AB - To study the optimum therapeutic modalities for treating human malignant brain tumors in vivo without ethical limitations, a model of heterotopic and another of orthotopic xenografting into nude mice were developed. For the first implantation, 11 human high-grade gliomas and 4 low-grade tumors were microsurgically grafted on epigastric vessels. The 11 high-grade gliomas, but no low-grade tumors, were established into nude mice. Afterwards, all these mouse adapted gliomas which grafted into other nude mice developed. Introduction of a microcatheter into the femoral artery or vein permitted infusion for magnetic resonance imaging (MRI) and treatment. A bladder catheter setting and electrode implantation allowed urine sampling and ECG or EEG recording. Thus, the most important parameters of chemo- and radiotherapy to destroy a maximum number of malignant cells or to inhibit their divisions and the hosts reactions to treatment can be studied. The human gliomas transplanted onto the mouse brain infiltrated the host brain at great distances from the tumor, as in human patients. So, this second implantation constitutes a representative model of the evolution of human gliomas, and allows the study of malignant cell migration in the brain before, during and after treatment determined with the heterotopic model and to appreciate the tolerance of the colonized brain to these treatments. Echography and MRI allowed us to follow the macroscopic evolution with or without treatment of the malignant brain tumors transplanted onto mouse brains. It should now be possible to undertake the same clinical studies on patients after appropriate consideration of ethical and scientific constraints. PMID- 12763242 TI - A novel method for monitoring surface membrane trafficking on hippocampal acute slice preparation. AB - Protein trafficking has attracted considerable attention as a potential regulator of neuronal plasticity. Therefore, it is of interest to study the mechanism involved in protein trafficking in experimental paradigms commonly used in this context. Here, we present a method for cell surface protein biotinylation in the acute hippocampal slice, the most commonly used preparation for electrophysiological recordings. We validated this procedure with two previously characterized cell surface receptors, glutamate receptor subunit A (GluR A) and the transferrin receptor (TfR). We observed a glutamate-dependent increase in the degradation of surface GluR A, whereas the TfR did not show significant degradation in the time window used. In addition, the presented method offers the opportunity to study processes such as internalisation and recycling, and can also be applied to examine the effect of normal and pathological patterns of activity on membrane protein trafficking in commonly used preparations for electrophysiological recordings. PMID- 12763244 TI - Video recording system for the measurement of eyelid movements during classical conditioning of the eyeblink response in the rabbit. AB - Classical conditioning of the eyeblink response in the rabbit is a popular model for studying the neural substrates of associative learning. Most of the common eyeblink recording techniques require invasive procedures. To perform experiments in intact animals, a non-invasive, high-speed video recording system was developed to measure eyeblink responses of rabbits during classical conditioning experiments. Besides being non-invasive, this method does not require excessive restraint of the animal. The PC-based system combines a Pulnix video camera with National Instruments video capture and timing hardware to control the experiment and acquire images of the peri-ocular region. The software developed for controlling these experiments also detects the eyeblink by measuring the movement of the upper and lower eyelids, the area of the exposed surface of the eye, and head movements in the sagittal plane. The time resolution of this relatively inexpensive system is 8.33 ms, and at a working distance of 0.8 m, it can detect movements as small as 0.11 mm. PMID- 12763243 TI - Western blotting with diaminobenzidine detection for the diagnosis of congenital disorders of glycosylation. AB - Congenital disorders of glycosylation (CDG) are a growing group of genetic disorders caused by a deficient assembly or processing of glycoproteins. Our aim was to improve a western blotting detection procedure previously described and to assess the efficiency of this procedure for CDG screening, using isoelectric focusing (IEF) as the reference method. We analysed transferrin and haptoglobin in serum from 12 patients with CDG-Ia, 3 patients with CDG-X and 95 healthy paediatric controls. These proteins were also studied in dried blood spot samples. Reference values for our paediatric population were established. No differences (Mann-Whitney test) were observed in the percentage of low molecular weight transferrin and haptoglobin fractions according to sex and age of the controls. Densitometric analysis showed a high percentage of the less sialylated fractions of glycoproteins in all CDG-Ia patients and normal values in the CDG-X patients. In conclusion, western blotting with diaminobenzidine detection is a simple and sensitive procedure to screen for CDG, either in serum or blood spot samples. Densitometric analysis and the establishment of reference values might improve the detection of subtle changes in the glycosylation of proteins. PMID- 12763245 TI - Tongue protrusion: a simple test for neurological recovery in rats following focal cerebral ischemia. AB - A simple tongue protrusion (TP) test is described for rats following focal ischemia induced by middle cerebral artery occlusion (MCAO). MCAO resulted in a dramatic decrease in TP that correlated with a concomitant decline in neurological performance in standard 5- and 20-point tests and deficits in performance in the Morris water maze and the accelerating rotarod. TP values also correlated with infarct size at 7 and 24 days following MCAO. This simple and inexpensive test, that monitors the ability of rats to lick food out of a glass tube, is easily administered, can be administered frequently without changing baseline performance, is not susceptible to behavioral compensation and should not interfere with other tests used concurrently to evaluate neurological deficit. The TP test may, therefore, serve as a useful addition to the battery of tests commonly used to assess neurological damage in rats, particularly in models of stroke. PMID- 12763246 TI - Determination of information flow direction among brain structures by a modified directed transfer function (dDTF) method. AB - A modification of directed transfer function-direct DTF-is proposed for the analysis of direct information transfer among brain structures on the basis of local field potentials (LFP). Comparison of results obtained by the analysis of simulated and experimental data with a new dDTF and DTF method is shown. A new measure to estimate direct causal relations between signals is defined. The present results demonstrate the effectiveness of the new dDTF method and indicate that the dDTF method can be used to obtain the reliable patterns of connections between various brain structures. PMID- 12763247 TI - Alternating-pulse iontophoresis for targeted cutaneous anesthesia. AB - In studies of sensory contributions to motor control, it may be advantageous to temporarily reduce the sensitivity of specific sensory systems. This article details a method for non-invasively inducing cutaneous anesthesia, leaving proprioceptive and motor functions intact. This method, called alternating-pulse iontophoresis, differs from conventional direct-current (DC) iontophoretic drug delivery in that adjacent drug delivery electrodes are stimulated out-of-phase. The total current delivered at any instant is then less than that produced during a comparable DC application, while the uniformity of drug delivery is expected to improve. Effective delivery of local anesthetics to the cutaneous foot soles by alternating-pulse iontophoresis was demonstrated using cutaneous pressure sensory threshold levels (STL's) assessed with Semmes-Weinstein monofilaments (arbitrary units of perceived force, or a.u.). Thirteen of 16 healthy subjects achieved a level of anesthesia greater than or equal to that normally associated with clinical peripheral sensory neuropathy. Average STL's measured prior to the anesthesia procedure were 4.00 a.u. ( approximately 10 mN). Immediately following the procedure, STL's were elevated to an average of 5.40 a.u. ( approximately 246 mN) and averaged 4.97 a.u. ( approximately 92 mN) after 50 min of standing. A number of research and clinical applications for this technique are suggested. PMID- 12763248 TI - Impaired cortical energy metabolism but not major antioxidant defenses in experimental bacterial meningitis. AB - The loss of soluble brain antioxidants and protective effects of radical scavengers implicate reactive oxygen species in cortical neuronal injury caused by bacterial meningitis. However, the lack of significant oxidative damage in cortex [J. Neuropathol. Exp. Neurol. 61 (2002) 605-613] suggests that cortical neuronal injury may not be due to excessive parenchymal oxidant production. To see whether this tissue region exhibits a prooxidant state in bacterial meningitis, we examined the state of the major cortical antioxidant defenses in infant rats infected with Streptococcus pneumoniae. Adenine nucleotides were co determined to assess possible changes in energy metabolism. Arguing against heightened parenchymal oxidant production, the high NADPH/NADP(+) ratio ( approximately 3:1) and activities of the major antioxidant defense and pentose phosphate pathway enzymes remained unchanged at the time of fulminant meningitis. In contrast, cortical ATP, ADP and total adenine nucleotides were on average decreased by approximately 25%. However, energy depletion did not lead to a significant decrease in adenylate energy charge (AEC). ATP depletion was likely a consequence of metabolic degradation, since it correlated with both the loss of total adenine nucleotides and accumulation of purine degradation products. Furthermore, the loss of ATP and decrease in AEC correlated significantly with the extent of neuronal injury. These results strongly suggest that energy depletion rather than parenchymal oxidative damage is involved in the observed cortical neuronal injury. PMID- 12763249 TI - NMDA induces NOS 1 translocation to the cell membrane in NGF-differentiated PC 12 cells. AB - Glutamatergic-mediated nitric oxide (NO) production occurs via the N-methyl-D aspartic acid (NMDA) postsynaptic density protein 95 (PSD95)-neuronal nitric oxide synthase (NOS1) ternary complex. To determine whether NOS1 is targeted to the membrane subsequent to NMDA receptor activation, we examined the effect of NMDA on NOS1 subcellular localization in nerve growth factor (NGF) differentiated PC12 cells. No effect on cell viability was observed using a range of NMDA concentrations from 500 to 1000 microM. Within 3 min of stimulation with 750 microM NMDA, increased cytoplasmic NOS1 immunostaining was observed with rapid membrane staining thereafter. This was inhibited by NMDAR inhibition with MK801. This observation was confirmed using subcellular fractionation and immunoblotting. Using 4, 5-diaminofluorescein diacetate (DAF2-DA) staining and a diazotization assay, concurrent NO production was observed. When PC 12 cells were co-treated with either NMDA and N(6)-nitro-L-arginine methyl ester hydrochloride (L-NAME) or (5R, 10S)-(+)-5-methyl-10, 11-dihydro-5H-dibenzo [a, d] cyclohepten 5, 10-imine hydrogen maleate (MK-801), nitric oxide (NO) generation was inhibited. Stimulation in a calcium-free medium did not increase NO levels. Although no evidence of cytotoxicity was observed utilizing either the MTT assay or measures of apoptosis within the maximal interval of NOS1 translocation, cell viability was reduced following 10 h of continuous NMDA exposure. While it has been shown that NMDA triggers NOS1 activation, these results indicate that NMDAR activation also mediates NOS1 targeting to the membrane. Our data validate that NGF-differentiated PC12 cells may be employed as a useful in vitro model to further study the regulation of NOS1 subsequent to NMDAR activation. PMID- 12763250 TI - Chronic corticotropin-releasing hormone and vasopressin regulate corticosteroid receptors in rat hippocampus and anterior pituitary. AB - Corticotropin-releasing hormone (CRH) and vasopressin (AVP) participate in the endocrine, autonomic, immunological and behavioral response to stress. CRH and AVP receptors are found in hippocampus and anterior pituitary, where mineralocorticoid (MR) and glucocorticoid (GR) receptors are abundant. We investigated the possible influence of CRH and AVP on the regulation of MR and GR in both tissues. CRH, AVP, or their antagonists were administered to adrenalectomized rats substituted with corticosterone, to avoid interference with adrenal secretion. Repeated i.c.v. oCRH injections (10 microgram) for 5 days significantly decreased MR and GR mRNA in hippocampus and MR mRNA in anterior pituitary. AVP significantly increased both corticosteroid receptor mRNAs, as repeated i.c.v. injections (5 microgram) for 5 days in hippocampus, and as continuous i.c.v. infusion (10 ng/h/5 days) in anterior pituitary. The i.c.v. infusion of 5 or 10 microgram/day of the alpha-helical CRH antagonist during intermittent restraint stress (5 days), induced a significant decrease in hippocampal MR binding. In anterior pituitary, 5 microgram/day significantly decreased MR binding, while 10 microgram/day significantly increased GR binding. Under the same conditions of stress, the infusion of 15 microgram/day of the vasopressin V1a/1b receptor antagonist [dP Tyr (Me)(2)AVP] significantly increased MR and GR binding in hippocampus and anterior pituitary; 5 microgram/day significantly decreased pituitary MR binding. Our results show that CRH and AVP regulate MR and GR in hippocampus and anterior pituitary. This reveals another important function of CRH and AVP, which could be relevant to understand stress adaptation and the pathophysiology of stress-related disorders like major depression. PMID- 12763251 TI - Immunohistochemical characterization of cholecystokinin containing neurons in the rat basolateral amygdala. AB - Specific neuronal populations in the basolateral amygdala (ABL) exhibit immunoreactivity for distinct neuropeptides and calcium-binding proteins. In the present study, immunohistochemical techniques were used to analyze neurons in the rat ABL that contain cholecystokinin (CCK). Some pyramidal projection neurons in the anterior subdivision of the basolateral nucleus exhibited low levels of CCK immunoreactivity in rats that received injections of colchicine to interrupt axonal transport; staining was concentrated in the axon initial segments of these cells. High levels of CCK immunoreactivity were observed in two subpopulations of nonpyramidal interneurons in all nuclei of the ABL: (1) type L neurons (characterized by large somata and thick dendrites), and (2) type S neurons (characterized by small somata and thin dendrites). Dual-labeling immunofluorescence studies using confocal laser scanning microscopy revealed that many (30-40%) type L CCK+ interneurons exhibited immunoreactivity for calbindin (CB), but not for parvalbumin (PV), calretinin (CR), or vasoactive intestinal polypeptide (VIP). In contrast, there was extensive colocalization of CR and VIP with CCK in type S neurons, but no significant colocalization with CB or PV. In addition, the majority of CR and VIP interneurons exhibited colocalization of both neurochemicals. Collectively, the results of this and previous studies indicate that there are at least four distinct interneuronal subpopulations in the ABL: (1) PV+ neurons (the great majority of which are CB+); (2) SOM+ neurons (many of which are CB+ and NPY+); (3) large CCK+ neurons (some of which are CB+); and (4) small bipolar/bitufted neurons that exhibit various amounts of colocalization of CCK, VIP, and CR. PMID- 12763252 TI - Locations and morphologies of sympathetically correlated neurons in the T(10) spinal segment of the rat. AB - We precisely localized and morphologically characterized sympathetically correlated neurons in the acutely transected spinal cord of the rat. We have shown that these neurons are likely members of the spinal networks that generate sympathetic activity after spinal cord transection. In humans with injured spinal cords, these networks are responsible for hypertensive crises that occur in response to ordinarily innocuous stimuli. We recorded from neurons in the dorsal horn of the T(10) spinal segment of anesthetized rats after acute spinal cord transection at C(2). Neurons with activities closely correlated to renal sympathetic nerve activity (RSNA) were considered to be putative components of spinal sympathetic systems. These neurons had receptive fields on the left flank and abdomen. After characterizing their ongoing activities, receptive fields, and degrees of correlation with RSNA, we juxtacellularly labeled neurons with biotinamide and subsequently reconstructed their somas and dendrites histologically. Confirming our earlier studies, sympathetically correlated neurons were found in dorsal horn laminae III, IV, and V. For the first time, we also identified sympathetically correlated neurons in laminae I and II. The dendrites of all sympathetically correlated neurons projected to multiple lamina. By virtue of the positions of their somas and the broad projections of their dendrites, we concluded that sympathetically correlated neurons may receive direct input both from supraspinal systems and from nociceptive and non nociceptive primary afferents. PMID- 12763253 TI - Stimulation of BIT induces a circadian phase shift of locomotor activity in rats. AB - Circadian rhythms of mammals are generated by a circadian oscillation of master pacemaker genes in the suprachiasmatic nucleus of the hypothalamus (SCN), and entrained by environmental factors such as 24-h light-dark cycles. We have previously shown that light exposure during the dark period enhanced tyrosine phosphorylation of brain immunoglobulin-like molecule with tyrosine-based activation motifs (BIT) in the rat SCN. To elucidate the functional roles of BIT in the circadian clock, we stimulated BIT using an anti-BIT monoclonal antibody (mAb) 1D4, which reacts with its extracellular region and induces phosphorylation of its intracellular tyrosine residues. Administration of mAb 1D4 into the third cerebral ventricle induced tyrosine phosphorylation of BIT in the SCN. Behavioral analyses showed that the SCN-injection of the antibody at CT15 induced a phase delay of the circadian rhythm of locomotor activity, and that at CT20 induced a phase advance. Pretreatment with MK801, a non-competitive antagonist of NMDARs, diminished the 1D4-induced phase shift at CT20, but not at CT15. These results suggest that BIT is involved in the entrainment of circadian rhythms through the function of NMDARs and non-NMDARs. PMID- 12763254 TI - Differential and irreversible CNS ontogenic reduction in maximal MK-801 binding site number in the NMDA receptor after acute hypoxic hypoxia. AB - CNS exposure to hypoxia impairs excitatory and inhibitory neurotransmission. Our aim was to determine variations induced by normobaric acute hypoxic hypoxia (8% O(2) for 60 min) on the NMDA receptor complex, as well as their potential reversibility after normoxic recovery. To this end, [3H]MK-801 binding assays to a synaptic membrane fraction isolated from chick optic lobes were performed. Previous studies throughout development had disclosed a characteristic age dependent pattern. Results at embryonic day (ED) 12 and 18 indicated two distinct MK-801 binding sites. Hypoxic treatment failed to alter either the high affinity site dissociation constant (K(d)) or its maximal binding capacity (B(max)), whereas the low affinity site B(max) was significantly decreased (50% and 30% at ED12 and 18, respectively), without alteration in its K(d) values. Hypoxic embryos restored for 48 h at ED12 to normoxic conditions displayed unchanged MK 801 binding reduction, unlike those treated likewise at ED18 whose values fully recovered control levels. To conclude, hypoxic treatment reduces low affinity MK 801 B(max) in the NMDA receptor which proves irreversible up to ED12. Such early neuronal vulnerability may be due to post-transcriptional changes, to endocytosis followed by receptor degradation, or alternatively to cell death. PMID- 12763256 TI - Periaqueductal gray-evoked dorsal root reflex is frequency dependent. AB - The dorsal root reflex (DRR) is an antidromic action potential originating in the spinal cord that propagates toward the periphery. Given that both GABA(A) and 5 HT(3) receptors are involved in the generation of DRRs and stimulation of the periaqueductal gray (PAG) can induce the release of GABA and serotonin within the spinal cord, we investigated the modulation of DRRs by the PAG descending system. The central end of the cut left L5 dorsal root in adult Sprague-Dawley rats was tested with single fiber recording. Stimulating electrodes were placed in the PAG, sciatic nerve, or transcutaneously across hindpaws. Fifty-seven DRRs were recorded for the effect of PAG stimulation in 19 rats, and 51 DRRs from 26 rats and nine DRRs from seven rats were recorded for an effect of ipsilateral and contralateral peripheral stimulation, respectively. The results were expressed as a percentage of the number of DRRs over the number of stimuli. PAG stimulation at 0.2, 0.5, 5, 20, and 50 Hz produced ratio's of 113.16+/-9.84, 114.54+/-12.22, 24.6+/-3.23, 17.77+/-4.76, and 12.62+/-3.44 (%), respectively. Stimulation at ipsilateral peripheral nerve evoked DRRs of 103.26+/-8.93, 95.27+/-10.57, 37.66+/ 7.55, 11.32+/-4.96, and 5.32+/-3.82 (%), respectively. Stimulation of the contralateral peripheral nerve evoked DRRs of 90.88+/-15.59, 44.30+/-10.77, 6.29+/-1.63, 0.45+/-0.19, and 0.29+/-0.15 (%), respectively. Transection at the thoracic spinal level completely eliminated PAG-induced DRRs. In conclusion, both PAG and peripheral stimulation produced DRRs in a frequency dependent manner. Stimulus intensity has no significant effect on DRRs. PMID- 12763255 TI - Effect of denervation of the locus coeruleus projections by DSP-4 treatment on [3H]-raclopride binding to dopamine D(2) receptors and D(2) receptor-G protein interaction in the rat striatum. AB - Changes in the control of dopaminergic neurotransmission by noradrenergic locus coeruleus (LC) projections has been implicated in such disorders as depression, drug addiction, and Parkinson's disease. In the present study, the effect of DSP 4, a neurotoxin highly selective for LC projections, on D(2) receptor abundance as assessed by [3H]-raclopride binding in the striatum was studied in rats after administration in doses of 10 and 50 mg/kg either 3 days or 1 month before decapitation. Three days after DSP-4 the levels of noradrenaline in the frontal cortex were dose-dependently reduced; after 1 month, noradrenaline levels were lowered only by the higher dose. DOPAC levels were dose-dependently reduced in the frontal cortex and striatum 3 days but not 1 month after DSP-4 treatment. Cortical 5-HIAA levels were reduced 3 days but not 1 month after DSP-4. The apparent number of D(2) receptor binding sites in the striatum was higher 1 month after either dose of DSP-4. DSP-4 treatment had no effect on [3H]-raclopride binding affinity, the ability of dopamine (DA) to compete with [3H]-raclopride binding and to activate [35S]GTPgammaS binding or on the binding affinities of GDP and [35S]GTPgammaS for corresponding G proteins 1 month after administration of the neurotoxin. These data suggest that after administration of DSP-4, short term reduction in DA and 5-HT metabolism occurs. Subsequently, an upregulation of D(2) receptor binding sites develops in the striatum even after a minor denervation of the LC projections. Thus, alterations in the LC projection systems elicit lasting adaptive changes in DA-ergic neurotransmission that can serve as a substrate for psychiatric disorders. PMID- 12763257 TI - Expression of brain-derived neurotrophic factor (BDNF) and inducible nitric oxide synthase (iNOS) in rat astrocyte cultures treated with Levetiracetam. AB - The aim of the present study was to investigate the effects of Levetiracetam, a new antiepileptic drug, on the synthesis of brain-derived neurotrophic factor (BDNF) and inducible nitric oxide synthase (iNOS) in rat cortical astrocyte cultures. The astrocytes were treated for 48 h with different concentrations of Levetiracetam and the expression of BDNF and iNOS was analyzed by immunostaining and immunoblotting analyses. We observed that Levetiracetam is able to stimulate expression of both BDNF and iNOS in a concentration-dependent manner on rat cortical astrocyte cultures. For the BDNF, this effect appears at very low concentrations (1 and 10 microgram/ml), while expression of iNOS appears only at higher dosages (50 microgram/ml). We conclude that Levetiracetam might exert neuroprotective effects, at least in part, via stimulation of neurotrophic factors, thus reducing the extent of inflammation and neuronal death under pathological conditions such as epilepsy. PMID- 12763258 TI - Ascorbic acid prevents water maze behavioral deficits caused by early postnatal methylmalonic acid administration in the rat. AB - Methylmalonic acidemia consists of a group of inherited neurometabolic disorders biochemically characterized by accumulation of methylmalonic acid (MA) and clinically by progressive neurological deterioration whose pathophysiology is not yet fully established. In the present study we investigated the effect of chronic administration (from the 5th to the 28th day of life) of methylmalonic acid (MA) on the performance of adult rats in the Morris water maze task. MA doses ranged from 0.72 to 1.67 micromol/g of body weight as a function of animal age; control rats were treated with the same volume of saline. Chronic postnatal MA treatment had no effect on body weight and in the acquisition of adult rats in the water maze task. However, administration of MA provoked long lasting reversal learning impairment in this task. Motor activity, evaluated by the swim speed in the maze, was not altered by MA administration, indicating no deficit of locomotor activity in rats injected with the metabolite. We also determined the effect of ascorbic acid administered alone or combined with MA on the same behavioral parameters in order to test whether free radicals might be responsible for the behavioral changes observed in MA-treated animals. Ascorbic acid was able to prevent the behavioral alterations provoked by MA. Moreover, the in vitro exposure of hippocampal and striatal preparations to MA revealed that the acid significantly reduced total radical-trapping antioxidant potential (TRAP) and total antioxidant reactivity (TAR) in the striatum, but not in the hippocampus. Furthermore, MA increased the thiobarbituric acid-reactive substances (TBA-RS) measurement in both structures. These data indicate that oxidative stress might be involved in the neuropathology of methylmalonic acidemia and that early MA administration induces long-lasting behavioral deficits, which are possibly caused by oxygen reactive species generation. PMID- 12763260 TI - Hypocretinergic facilitation of synaptic activity of neurons in the nucleus pontis oralis of the cat. AB - The present study was undertaken to explore the neuronal mechanisms of hypocretin actions on neurons in the nucleus pontis oralis (NPO), a nucleus which plays a key role in the generation of active (REM) sleep. Specifically, we sought to determine whether excitatory postsynaptic potentials (EPSPs) evoked by stimulation of the laterodorsal tegmental nucleus (LDT) and spontaneous EPSPs in NPO neurons are modulated by hypocretin. Accordingly, recordings were obtained from NPO neurons in the cat in conjunction with the juxtacellular microinjection of hypocretin-1 onto intracellularly recorded cells. The application of hypocretin-1 significantly increased the mean amplitude of LDT-evoked EPSPs of NPO neurons. In addition, the frequency and the amplitude of spontaneous EPSPs in NPO neurons increased following hypocretin-1 administration. These data suggest that hypocretinergic processes in the NPO are capable of modulating the activity of NPO neurons that receive excitatory cholinergic inputs from neurons in the LDT. PMID- 12763261 TI - 5alpha-Reduced androgens block estradiol-BSA-stimulated release of oxytocin. AB - In this study we test the postulate that estradiol conjugated to bovine serum albumin (E-BSA) acts via receptors for the steroid-binding protein sex hormone binding globulin (SHBG) by attempting to block E-BSA-stimulated release of oxytocin with two antagonists of SHBG receptor actions: the 5alpha-reduced androgens dihydrotestosterone (DHT) and 3alpha-diol. Simultaneous superfusion with either DHT or 3alpha-diol significantly blocked E-BSA-stimulated release of oxytocin. We also found that a wide range of free 17beta-estradiol was unable to stimulate oxytocin release, suggesting that E-BSA stimulates receptors other than those for free estradiol to release oxytocin, perhaps SHBG receptors. PMID- 12763259 TI - Paraquat induces long-lasting dopamine overflow through the excitotoxic pathway in the striatum of freely moving rats. AB - The herbicide paraquat is an environmental factor that could be involved in the etiology of Parkinson's disease. We have previously shown that paraquat penetrates through the blood-brain barrier and is taken up by neural cells. In this study, we examined the in vivo toxic mechanism of paraquat to dopamine neurons. GBR-12909, a selective dopamine transporter inhibitor, reduced paraquat uptake into the striatal tissue including dopaminergic terminals. The subchronic treatment with systemic paraquat significantly decreased brain dopamine content in the striatum and slightly in the midbrain and cortex, and was accompanied by the diminished level of its acidic metabolites in rats. When paraquat was administered through a microdialysis probe, a transitory increase in the extracellular levels of glutamate, followed by long-lasting elevations of the extracellular levels of NO(x)(-) (NO(2)(-) plus NO(3)(-)) and dopamine were detected in the striatum of freely moving rats. This dopamine overflow lasted for more than 24 h after the paraquat treatment. Dopamine overflow was inhibited by N(G)-nitro-L-arginine methyl ester, dizocilpine, 6,7-dinitroquinoxaline-2,3-dione and L-deprenyl. The toxic mechanism of paraquat involves glutamate induced activation of non-NMDA receptors, resulting in activation of NMDA receptor channels. The influx of Ca(2+) into cells stimulates nitric oxide synthase. Released NO would diffuse to dopaminergic terminals and further induce mitochondrial dysfunction by the formation of peroxynitrite, resulting in continuous and long-lasting dopamine overflow. The constant exposure to low levels of paraquat may lead to the vulnerability of dopaminergic terminals in humans, and might potentiate neurodegeneration caused by the exposure of other substances, such as endogenous dopaminergic toxins. PMID- 12763263 TI - Influence of predation pressure on the escape behaviour of Podarcis muralis lizards. AB - Relationships between predator avoidance behaviour and predation pressure were investigated in the wall lizard, Podarcis muralis. The wariness of lizards belonging to high (1185m) and low elevation (308m) populations under two different predation pressure levels was compared. Wall lizards belonging to the lowland population experienced greater predation pressure than those belonging to the highland population. Lizards belonging to the population under higher predation pressure had higher frequency of refuge use, and had longer flight initiation distances (i.e. the distance lizards allowed the observer to approach before fleeing). In contrast, neither the distance fled (i.e. the total distance they fled in one continuous movement from the lizard's initial position until hiding or stopping at a safe distance) nor the distance to the nearest refuge were significantly different between populations. Escape responses were independent of ambient temperature in the lowland population, but animals belonging to the highland population had longer flight initiation distances when the ambient temperatures were higher. These findings suggest that predator avoidance behaviour may vary with predation pressure. PMID- 12763262 TI - N(2)O stimulates NOS enzyme activity in C57BL/6 but not DBA/2 mice. AB - Exposure to 70% N(2)O produces a prominent antinociception in C57BL/6 mice but not DBA/2 mice. N(2)O exposure also increases conversion of [14C]L-arginine to [14C]L-citrulline in homogenates prepared from whole brains of C57BL/6 mice; there was no such increase in NOS activity in the DBA/2 whole brain. A differential N(2)O effect on brain NOS in these inbred strains might explain why the C57BL/6 but not DBA/2 mice are responsive to N(2)O antinociception. PMID- 12763264 TI - Characterization of NFH-LacZ transgenic mice with the SHIRPA primary screening battery and tests of motor coordination, exploratory activity, and spatial learning. AB - NFH-LacZ transgenic mice express a fusion protein between a truncated form of the endogenous neurofilament of heavy molecular weight and the complete E. coli beta galactosidase. NFH-LacZ transgenic mice could be distinguished from controls in the SHIRPA neurological battery by the appearance of action tremor and hindlimb clasping and a lower body weight. Despite normal exploratory activity and spatial learning, NFH-LacZ transgenic mice were deficient in stationary beam, coat hanger, and rotorod tests of motor coordination. These results are concordant with neuropathological findings in spinal motoneurons and the cerebellum and indicate that despite the absence of paralysis, these transgenic mice may serve as an experimental model of the early stage of amyotrophic lateral sclerosis. PMID- 12763265 TI - Empirical and theoretical implications of additivity between reinstatement and renewal after interference in causal learning. AB - An experiment explored the effects of context change (renewal) and presentation of the outcome (reinstatement) upon retroactive interference in causal learning. An interference task was used where a sequential relationship between the name of a food and two different outcomes was established (A+|A*). Renewal and reinstatement effects appeared as partial attenuation of retroactive interference. The combination of both treatments produced complete recovery of first-learned information (A+). This complete reversal on the estimations of participants suggests that renewal and reinstatement effects within this paradigm imply partial recovery of the first-learned information, rather than response at a chance level emitted by confused participants. On the other hand, additivity between reinstatement and renewal suggests that reinstatement may be due to the combined effect of different underlying processes. PMID- 12763266 TI - Dispersion movements in ants: spatial structuring and density-dependent effects. AB - This paper examines whether the characteristics of individual dispersion movements in ants are changed when workers are moving solitarily or in a group. We analyzed the trajectories of workers of the species Messor sancta moving solitarily or in groups of different size (5, 10, 15 individuals), tested for density-dependent effects on their trajectory characteristics and investigated through resampling techniques whether ants are able to spatially structure their movements through direct (e.g. contact) or indirect (pheromone deposited on the ground) interactions. In addition to group size, the effects of the nutritional state of the colony and of the state of the area on which ants were dispersing were also examined. Solitary ants moved faster and had more sinuous trajectories than ants moving in a group. We found however no significant differences in trajectory characteristics between groups of different size. Whatever the group size, ants from starved colonies moved more slowly and had more direct trajectories than their counterpart coming from fed colonies. On the other hand, the state of the area on which ants were moving had no direct significant effect on dispersion movement. Ants dispersing in a group moved independently and did not coordinate their movements through direct or indirect interactions. However, the geometry of their path was changed not only through the effect of random encounters with other workers but also through an active modification of their movement when they perceived directly or indirectly the presence of nearby workers. PMID- 12763267 TI - Effects of haloperidol on rat behavior and density of dopaminergic D2-like receptors. AB - The present work shows the effects of a typical neuroleptic drug (haloperidol, HAL) on rat behavior (catalepsy and locomotor activity) and dopaminergic D2-like receptor densities in the hippocampus and striatum. Male Wistar rats (2-3 months old) were treated daily for 30 days with HAL (0.2 or 1mg/kg, intraperitoneally (i.p.)). At the end of treatment and 1h or 1, 3, 7 and 15 days after drug withdrawal, animals were subjected to behavioral tests and sacrificed afterwards for binding assays. The results showed that behavioral effects with both doses were significant only 1h and 1 day after withdrawal, and similar to controls at the third day. An up-regulation of D2 receptors was observed in the striatum (28% increase) but not in the hippocampus after 24h HAL (1mg/kg) withdrawal. However, an up-regulation was seen in both areas (1mg/kg) 3 days after drug withdrawal (58 and 42% increases in the hippocampus and striatum, respectively), and continued after 7 days of withdrawal only in the striatum (43 and 49% for the doses of 0.2 and 1mg/kg, respectively), suggesting the influence of dose, age, and time of drug withdrawal on these parameters. The up-regulation disappeared after 15 days of haloperidol withdrawal. Increases (72 and 140%) in constant dissociation values (K(d)) values were also observed 7 days after withdrawal. Results show differences on a time-basis between behavioral alterations and dopaminergic D2 receptors up-regulation. PMID- 12763268 TI - Timing of presentation of an audience: aggressive priming and audience effects in male displays of Siamese fighting fish (Betta splendens). AB - Studies of animal communication often underestimate the presence of individuals other than the signaller-receiver dyad. Signalling interactions often occur in the presence of non-participating individuals (audiences); the effect of these individuals upon the dynamics of interactions has been called the audience effect. Recent studies of fighting fish Betta splendens have shown that the presence of a male audience can increase aggression during interactions. However, in many of these studies males were allowed to see the audience prior to the interaction, thus such pre-exposure may have facilitated aggressive behaviour (aggressive priming). Here we present results of two experiments designed to examine the relative importance of priming and audience effects on the dynamics of aggressive interactions. Males that were pre-exposed showed higher levels of aggression during subsequent interactions regardless of the presence or absence of an audience. When only one of the interactants had been pre-exposed to the audience, the non-exposed male showed similar increases in aggressive behaviour, i.e. matching the level of aggression showed by his opponent. Taken together these results suggest that aggressive priming may have resulted in an over estimation of the audience effect in previous studies. The results still highlight the importance of social environment in determining the dynamics and outcomes of aggressive contests. PMID- 12763269 TI - The Roman high- and low-avoidance rats respond differently to novelty in a familiarized environment. AB - The purpose of the present study was to investigate the response to novelty in Roman high- and low-avoidance rats under non-stressful conditions. To reduce fear, a procedure of repetitive placing in the experimental chamber consisting of start, screen, and tunnel zones was applied. Each animal was placed in the experimental chamber daily for a 6min period. The first 11 sessions were the habituation sessions. In the 12th session, the novelty was introduced into the screen and tunnel zones. The subsequent two sessions were conducted under novelty conditions. Behavioral activities such as walking, object contacts, time spent in given zones, and entering the tunnels were measured. All the comparisons were made for two 3-min intervals, using a three-factor MANOVA, involving 2(sex)x2(subline)x8(3-mininterval). All subjects increased time spent in the tunnel zone, but RHA/Verh rats responded to a greater extent, especially the males. All subjects spent shorter times in the screen zone, but the RHA/Verh rats responded to a much greater extent. RHA/Verh rats, especially males responded with a substantial increase of time spent inside the tunnels. All subjects responded with an increased amount of object contacts. In general the RHA/Verh subjects showed a more pronounced response to novelty, as evidenced by a significant shift toward the tunnel zone. They spent more time in this zone than their RLA/Verh counterparts. Among the RLA/Verh rats, males tended to behave similarly to RHA/Verh rats, especially during the second 3min interval of session "12." The differences between the rat lines obtained in this study may be attributed to mechanisms specific to exploration, making them promising subjects to study the relationships between reactivity, novelty detection, adaptation, and environmental information processing. PMID- 12763270 TI - Upcoming food-pellet reinforcement alters rats' lever pressing for liquid sucrose delivered by a progressive-ratio schedule. AB - The present study investigated whether rats' responding for liquid-sucrose reinforcement delivered by a progressive-ratio (PR) schedule would be altered by the addition of food-pellet reinforcement available subsequent to the PR schedule. In Experiment 1, six rats lever pressed for 1% sucrose reinforcers delivered by a PR 3 schedule. In Experiment 2, six rats lever pressed for 5% sucrose delivered by a PR 5 schedule. In both experiments, baseline sessions consisted of 40min of exposure to the PR schedule. In the first treatment condition, a 25-min period of food-pellet reinforcement, delivered by a random interval 60-s schedule, immediately followed the initial 40min. In the second treatment condition, the 25-min period of food-pellet reinforcement became available when 10min elapsed without the subject completing a ratio on the PR schedule. Results from both experiments showed that upcoming food-pellet reinforcement increased the number of ratios subjects completed on the PR schedule. Portions of the present results represent a partial replication of results reported by Baron and Derenne [J. Exp. Anal. Behav. 73 (2000) 291], who used a similar procedure. They also augment a growing body of research on positive induction. PMID- 12763271 TI - Conflict as a determinant of rat behavior in three types of elevated plus-maze. AB - Three groups of rats were tested in different types of elevated plus-mazes, a normal one (two closed and two open arms), a totally closed one (four closed arms) and a totally open (four open arms). Closed arms were surrounded by 40-cm high wooden walls and open arms were surrounded by 0.5-cm high transparent Plexiglas ledges. As expected, in the closed maze rats explored equally all the arms, both in terms of time and frequency of entries, as well as in exploration of the extremities. Rats in the totally open maze also presented a similar pattern of exploration, that is, no significant differences were found between the results obtained with the closed and the open mazes in terms of central and extremities exploration. It is suggested that the typical behavior of rats in the conventional elevated plus-maze is caused by the contrasting characteristics of open and closed arms rather than by the physical aversive characteristics of the open arms per se. Results also confirm a prediction made by a computer model simulating rat exploratory behavior in virtual mazes, normal, totally open and totally closed. PMID- 12763272 TI - Rats have trouble associating all three parts of the time-place-event memory code. AB - The ability of animals to associate an event with predictable time and place information confers a major biological advantage. The current research uses a variety of procedures and paradigms (e.g. place preference, radial arm maze, Morris water maze, T-maze, go no-go) to show that rats, unlike pigeons [e.g. Anim Learn Behav 22 (1994) 143] do not readily make an event-time-place association. They do make associations between event-time and event-place information, however. These findings are in disagreement with Gallistel's (The Organization of Learning, MIT Press, Cambridge, MA ) theory that claims that animals automatically store a memory code that has these three pieces of information. The present research is in line with the work of others who also find that rats do not readily make daily time-place associations [Behav Processes 23 (1997) 232; Behav Processes 52 (2000) 11; Behav Processes 49 (2000) 21; Anim Learn Behav 28 (2000) 298]. An interesting finding that did emerge from the present research was that at least some rats can use a circadian timer to solve a time-of-day discrimination if the task is a go no-go discrimination. PMID- 12763273 TI - Advances in the detection of Alzheimer's disease-use of cerebrospinal fluid biomarkers. AB - The diagnosis of Alzheimer's disease (AD) is still made by excluding other disorders with a similar clinical picture. In addition, an analysis of symptoms and signs, blood analyses and brain imaging are the major ingredients of the clinical diagnostic work-up. However, the sensitivity of a clinical diagnosis using these instruments is unsatisfactory and disease markers with high sensitivity and specificity for AD would be a welcome supplement. Ideally, such markers should reflect the pathophysiological mechanisms of AD, that is, according to the currently predominant hypothesis mismetabolism of beta-amyloid and neurofibrillary degeneration. Among several, we have focused on three candidates that have been suggested to fulfil the requirements for biomarkers of AD: beta-amyloid42 (Abeta42), total tau (T-tau) and tau phosphorylated at various epitopes (P-tau). The cerebrospinal fluid (CSF) levels of these proteins reflect the metabolism of these proteins in the central nervous system. Only published articles using established ELISA methods for the quantification of these markers in CSF and preferably also presenting sensitivity and specificity figures have been included in this review. The number of patients included in the different studies varies; some having included only a few patients. Furthermore, diagnostic criteria vary and clinicopathological studies are scarce. However, there are some large studies, including even minor studies, and most have found reduced CSF levels of Abeta42 and increased CSF levels of T-tau in AD. The sensitivity and specificity of these measures are high for separation of AD patients from controls, but their specificity against other dementias is moderate. It increases if P-tau is added. An increasing number of studies suggest that supplementary use of these CSF markers, preferably in combination, adds to the accuracy of an AD diagnosis. PMID- 12763274 TI - An observational study of severe hypertriglyceridemia, hypertriglyceridemic acute pancreatitis, and failure of triglyceride-lowering therapy when estrogens are given to women with and without familial hypertriglyceridemia. AB - BACKGROUND: We assessed severe hypertriglyceridemia, hypertriglyceridemic acute pancreatitis, and failure of triglyceride-lowering therapy when estrogens were given to 56 women with and without familial hypertriglyceridemia. The 56 women had been consecutively referred to our center over a 3-year period because of triglycerides >400 mg/dl despite diet-drug treatment and/or a history of hypertriglyceridemic acute pancreatitis (AP). Of the 56 women, 17 had received estrogen replacement therapy (ERT), hormone replacement (HRT, n=6), or selective estrogen receptor modulators (SERM, n=1). METHODS: After study at entry, in 56 women (median age, 52 years), 36 with familial hypertriglyceridemia, to lower triglycerides, estrogens and SERMs (hormone treatment, HT) were stopped; a very low fat diet (<15% of calories), gemfibrozil (1.2-1.5 mg/day), and omega-3-fatty acid (4-12 g/day) were started, with restudy 2-4 weeks later. RESULTS: Of the 56 women, 24 (43%) were taking HT at entry, with median fasting triglycerides 1270 mg/dl in the HT group and 1087 mg/dl in the no-HT group. Seventeen women (30%) had a history of AP, nine of whom (53%) were/had been on HT at the development of AP. Significant positive correlates of triglycerides at entry in a stepwise regression model were hemoglobin A(1C) (partial r(2)=10.7%, p<0.05) and an interaction between estrogen use and familial hypertriglyceridemia (partial r(2)=15%, p=0.017). After 2-4 weeks on therapy, median triglycerides in the previous-HT group fell from 1270 to 284 mg/dl (p<0.0001) and in the no-HT group from 1087 to 326 mg/dl (p<0.0001). CONCLUSIONS: Before starting HT, to avoid HT induced hypertriglyceridemic AP and exacerbation of overt or covert familial hypertriglyceridemia, triglycerides must be measured. HT is contraindicated in women with preexisting hypertriglyceridemia (triglycerides> or =500 mg/dl). Triglyceride-lowering diets and drugs often fail in the presence of HT and/or poorly controlled diabetes mellitus, but commonly succeed when HT is stopped and diabetes mellitus is tightly controlled. PMID- 12763275 TI - Commentary. Secondary causes of hypertriglyceridemia and pancreatitis. PMID- 12763276 TI - Plasma homocysteine and its relationships with atherothrombotic markers in psoriatic patients. AB - BACKGROUND: Hyperhomocysteinemia may constitute an independent risk factor for cardiovascular disease and may promote atherothrombosis. Psoriasis is one of the diseases associated with increased atherothrombosis. The aim of the present study was to examine serum total homocysteine (tHcy) level and its relationships with atherothrombotic markers. METHODS: The study group included 30 patients with psoriasis (17 females and 13 males) with a mean age of 34.2 (age range: 27-40) and 30 sex and age matched healthy volunteers (15 females and 15 males) with a mean age of 36.7 (age range: 26-48). The concentrations of lipids, lipoproteins, acute phase reactants, tHcy and atherothrombotic markers [fibronectin, soluble vascular adhesion molecules-1 (sVCAM-1), soluble intercellular adhesion molecules 1 (sICAM-1), tissue plasminogen activator (tPA), plasminogen activator inhibitor 1 (PAI-1), autoantibodies against oxidized LDL (AuAb-oxLDL)] were determined. RESULTS: The mean levels of serum tHcy, fibrinogen, fibronectin, sICAM, PAI-1 and AuAb-oxLDL were increased in patients whereas tPA, vitamin B(12) and folate levels were decreased significantly. Increased levels of sVCAM were not statistically significant. tHcy levels were negatively correlated with vitamin B(12) (r=-0.40, P=0.027) and positively correlated with PAI-1 and AuAb-oxLDL levels (r=0.46, P=0.011; r=0.39, P=0.035, respectively). CONCLUSIONS: It was concluded that the increased homocysteine concentration and altered endothelial cell-mediated proteins associated with increased lipids and LDL oxidation may play an important role for the development of atherothrombotic complications with psoriasis. PMID- 12763277 TI - Rapid genotyping of paraoxonase 55 and 192 mutations by melting point analysis using real time PCR technology. AB - BACKGROUND: Paraoxonase (PON1) enzyme was identified as one of the components of HDL responsible for prevention of lipid peroxides accumulation in low-density lipoprotein (LDL). A triphasic phenotypic frequency distribution of PON1 activity was shown in the human population resulted by two nucleotide interchanges at residues 55 and 192. The paraoxonase isoforms have different effectiveness in hydrolysing lipid peroxides. METHODS: To date, genotyping for PON1 is mainly performed by PCR RFLP technique, that is time consuming and sensitive to contamination. We developed highly reliable single-step methods for genotyping both PON1 55 and 192 polymorphisms using LightCycler real time PCR technology based on fluorescence resonance energy transfer. After the ultrafast PCR, melting point analysis was performed and fluorescence intensity was monitored simultaneously with slow heating. RESULTS AND CONCLUSIONS: The observed melting temperatures in the PON1 55 and 192 melting point analyses characteristic to the oligonucleotides hybridised to the mutant and wild-type DNA were 57 degrees C, 61 degrees C and 51.5 degrees C, 57.5 degrees C, respectively. The temperature differences in melting points (4 degrees C and 6 degrees C, respectively) offer a powerful tool for rapid, reliable mutation detection for 55 and 192 polymorphisms even in routine diagnostic laboratories or large epidemiological studies. PMID- 12763278 TI - Red pepper attenuates cholesteryl ester transfer protein activity and atherosclerosis in cholesterol-fed rabbits. AB - BACKGROUND: The current study was conducted to examine the effect of red pepper supplementation on cholesteryl ester transfer protein (CETP) activity, along with its anti-atherosclerotic effect in cholesterol-fed rabbits. METHODS: Rabbits were fed a 1% cholesterol diet for 12 weeks, including a 1% red pepper powder supplement. RESULTS: The red pepper supplemented group exhibited significantly lower CETP activity than the control group during the experimental period (P<0.05). The total cholesterol, triglyceride (TG), LDL-C, VLDL-C, and VLDL-TG levels and atherogenic index (AI) were all significantly lower in the red pepper group than in the control group (P<0.05), whereas the HDL-C level was significantly higher in the red pepper group than in the control group during the experimental period (P<0.05). Furthermore, the red pepper supplementation increased the fecal TG excretion (P<0.05). Based on a morphological examination, the red pepper supplemented group exhibited fewer fat droplet deposits than the control group. CONCLUSIONS: The current results suggest that red pepper attenuates atherosclerosis, plus plasma CETP would appear to be a risk marker of atherosclerosis in cholesterol-fed rabbits. PMID- 12763279 TI - Evaluation of Pankrin, a new serum test for diagnosis of acute pancreatitis. AB - AIM: Evaluation of a new serum test for diagnosis of acute pancreatitis. PATIENTS AND METHODS: One hundred and sixty-three patients presenting with acute abdominal pain were included into the study. Acute pancreatitis was diagnosed by CT or ultrasound. Serum samples were taken 0-1 days, 2-3 days, and 4-5 days after onset of symptoms and C-reactive protein, lipase, elastase, and amylase were determined. As a further parameter, Pankrin, a newly available kit for the measurement of a mixture of elastase and other pancreatic secretory proteins was used. As control, serum from 558 apparently healthy blood donors was analysed. The receiver operator characteristics (ROC) and the areas under the curves (AUC) were calculated for each individual test. RESULTS: In Western blot analysis the antibodies of the Pankrin assay detected the majority of protein bands in human pancreatic juice. In blood donors, the median value of Pankrin was 88 U/ml (range 14-316 U/ml). In 16 from 163 patients with acute abdominal pain, acute pancreatitis was diagnosed and the median Pankrin level in samples collected on days 0-1 was 345 U/ml (range 220-518 U/ml, p<0.0001). In those patients with abdominal pain but without pancreatitis, the median was 116 U/ml (range 17-396 U/ml). The ROC-curves for amylase, lipase, elastase, and Pankrin from samples collected after 0-1 days were similar (area under the curves (AUC) >0.98). After 2-3 days, the AUC of all markers decreased (AUC 0.80-0.89) and after 4-5 days the AUC of Pankrin (0.85) was higher than all other parameters. CONCLUSION: In those patients with abdominal pain, who present several days after onset of pain, the new serum test for pancreatitis, Pankrin, could be of help to improve the diagnosis of pancreatitis. PMID- 12763281 TI - Gemfibrozil reduces release of tumor necrosis factor-alpha in peripheral blood mononuclear cells from healthy subjects and patients with coronary heart disease. AB - BACKGROUND: Inflammatory process plays an important role in the pathogenesis of coronary heart disease (CHD). With the growing use of gemfibrozil and other fibrates, their anti-inflammatory effects have been noted. But little is known about the effect of gemfibrozil on tumor necrosis factor (TNF)-alpha secretion in peripheral blood mononuclear cells (PBMC) from patients with coronary heart disease. METHODS: PBMC were obtained from CHD patients (n=16) and healthy controls (n=13). PBMC (2x10(6) cells/ml) were cultured in 24-well plates with or without Ang II (10(-8), 10(-7), 10(-6) mol/l), or Ang II (10(-6) mol/l) plus gemfibrozil (10(-6), 10(-5), 10(-4) mol/l). After 24-h incubation, the supernatants were separated, and TNF-alpha was measured by an enzyme-linked immunosorbent assay (ELISA). RESULTS: Spontaneous release of TNF-alpha was 299.2+/-110.7 pg/ml in PBMC from CHD patients and 179.3+/-78.2 pg/ml in PBMC from control subjects (P<0.05). Incubated with Ang II (10(-8), 10(-7), 10(-6) mol/l), TNF-alpha secretion was 307.7+/-141.8, 318.9+/-135.6, 328.6+/-123.9 pg/ml in PBMC from CHD patients, and 225.3+/-135.4, 224.1+/-141.0,218.7+/-134.8 pg/ml in PBMC from control subjects, respectively. Ang II did not significantly trigger TNF alpha secretion in both groups. Compared with that incubated with Ang II (10(-6) mol/l) alone, release of TNF-alpha intervened by gemfibrozil (10(-6),10(-5),10( 4) mol/l) decreased to 279.4+/-132.2, 268.0+/-132.7, 226.6+/-102.7 pg/ml in PBMC from CHD patients, and 177.6+/-94.4, 156.1+/-69.4, 105.3+/-52.7 pg/ml in the control group, respectively. Gemfibrozil (10(-5),10(-4) mol/l) significantly inhibited TNF-alpha secretion in both groups (P<0.05). CONCLUSIONS: Our data demonstrated that gemfibrozil reduced release of TNF-alpha in PBMC both from CHD patients and controls. This effect may partially be relevant to the clinical benefits of gemfibrozil in the treatment of dyslipidemia and atherosclerosis. PMID- 12763280 TI - Development of a point-of-care assay system for high-sensitivity C-reactive protein in whole blood. AB - BACKGROUND: C-reactive protein (CRP) is emerging as a potential risk predictor for future cardiovascular diseases (CVD). High sensitivity assays have been developed and applied for clinical purposes. METHODS: The fluorescence immunochromatographic assay was employed to detect and quantify CRP in whole blood. It consisted of a fluorescence (FL) antibody detector buffer, a test strip housed in a disposable cartridge, and a laser fluorescence scanner. Whole blood sample was mixed with detector, loaded onto a cartridge, incubated for 10 min, and the concentration of CRP was measured in a laser fluorescence scanner. The linearity, limit of detection (LOD), and performance of new assay system was tested and evaluated. The comparability of assay was examined with an automated reference method. RESULTS: With the new assay system, a reliable correlation of coefficient (r) was obtained between the ratio value (A(T)/A(C)) and a concentration of CRP in samples. The linearity fell in the range of 0-10 mg/l of CRP, and the analytical detection limit was 0.133 mg/l of CRP. The mean recovery of the control was 105.2% in a working range. The precision of the intra- and inter-assay in a range of 0.5-6 mg/l was CVs <6% and <8%, respectively. The new fluorescence immunochromatographic assay system correlated well with a traditional immunoturbidimetric assay for quantification of CRP concentration (r=0.955, N=90). CONCLUSION: The fluorescence immunochromatographic assay is fast, reliable, and a reproducible platform for point-of-care testing (POCT) of high-sensitive (hs)-CRP in whole blood. PMID- 12763282 TI - Characterization of copper in uterine fluids of patients who use the copper T 380A intrauterine device. AB - BACKGROUND: The copper intrauterine device (IUD) is a highly effective method of contraception that requires the dissolution of the copper into uterine cavity. However, there is little information about the amount and form of copper in the fluid and whether the presence of this element produces any change in the protein concentration. METHODS: Twenty-seven women were divided into three groups that had used IUD for about 6 months, 1 year and > or =3 years. The samples were collected during the proliferative phase (Pp), secretory phase (Sp) and menstruation (M). Square-wave anodic stripping voltammetry (SWASV), cyclic voltammetry (CV), high performance liquid chromatography (HPLC) and atomic absorption spectrometry (AAS) were used in this study. RESULTS: Total copper concentrations were between 3.9 and 19.1 micro g/ml. The mean and standard deviations were as follows: 6 months, 11.4+/-4.7 micro g/ml of copper; 1 year, 11.5+/-7.0 micro g/ml of copper; and 3 years, 6.2+/-1.5 micro g/ml of copper. Total proteins were quantified by measuring the area under the chromatographic peaks. The mean areas obtained with uterine fluid samples from women who used IUDs for 6 months, 1 year and 3 years were 290,013, 538,934 and 201,863 arbitrary units (AU), respectively. The control sample was only 22323. CONCLUSIONS: The amount of copper released from IUD, although high, is in the form of complexes with proteins. IUDs have a constant copper release for at least 6-12 months. Copper(I) was not detected in the fluid. Copper induces a change in the total protein concentration. The amount of copper released and the amount of proteins is slightly larger during the menstrual stage. PMID- 12763283 TI - Variations of whole blood viscosity using Rheolog-a new scanning capillary viscometer. AB - BACKGROUND: Whole blood viscosity (WBV) values identify subjects at high risk for initial or recurrent cardiovascular events. However, these measurements have been limited to specialized centers. A new type of viscometer, Rheolog, was designed to overcome the difficulties encountered in WBV measurements using the standard rotational viscometer in a clinical environment. METHODS: We evaluated the 14-day variability of WBV measured by Rheolog in a single-center study of 24 healthy male subjects aged 18-75 years. WBV was measured through an 11-h period on study days 1, 8, and 14. An additional fasting WBV test was performed on study days 3, 5, and 11. RESULTS: Average morning measurements were higher than afternoon measurements at all shear rates. Both inter- and intraindividual variations were higher in the morning than later in the day, but the differences between pooled mean values were not significant. Interindividual variations at fasting were higher than the pre-meal or overall variations. There was a small nonsignificant increase in mean viscosity following each meal. CONCLUSION: WBV measurements using Rheolog have potential for clinical application because of the convenience and low variability of measurements over time. PMID- 12763284 TI - Lipid peroxidation and osmotic fragility of red blood cells in sleep-apnea patients. AB - BACKGROUND: Obstructive sleep apnea (OSA) refers to the occurrence of episodes of complete or partial pharyngeal obstruction with oxyhemoglobin desaturation during sleep. These hypoxia/reoxygenation episodes may cause generation of reactive oxygen species. Reactive oxygen species are toxic to biomembranes and may lead to the peroxidation of lipids. We tested the hypothesis that obstructive sleep apnea is linked to increased oxidative stress and lipid peroxidation. In order to identify target tissue/cell damage, we studied the osmotic fragility of red blood cells. METHODS: Six subjects polysomnographically diagnosed as obstructive sleep apnea syndrome and 10 controls were included. After all subjects gave written informed consent, blood samples were collected in the morning between 08:00 and 09:00 a.m. following polysomnography. Blood samples were immediately transferred to the laboratory. Glutathione, lipid peroxidation and osmotic fragility of red blood cells were measured manually. RESULTS: Mean glutathione and lipid peroxidation concentrations of patients were not different than those of control subjects (105.6+/-38.6 U/g Hb and 3.1+/-2.3 nmol MDA/l vs. 100.6+/-62.1 U/g Hb and 3.2+/-2.8 nmol MDA/l, respectively). In both groups, osmotic fragility of red blood cells was not changed. CONCLUSION: The present study failed to support the hypothesis that obstructive sleep apnea is linked with increased oxidative stress and lipid peroxidation. PMID- 12763285 TI - Rules-based detection of discrepancies between TSH and free T4 results. AB - BACKGROUND: Analytical errors in clinical laboratory testing are unavoidable. Recent reports have suggested the idea of "physiological profiling" which uses several results from a given patient to identify clinically unlikely results. The objective of this study was to establish rules-based criteria for identifying physiologically unlikely TSH and free T(4) (fT(4)) results. METHODS: For a 30 month period, all samples with fT(4) concentrations >2 ng/dl and TSH concentrations >0.1 micro IU/ml were investigated. RESULTS: Among 7918 plasma samples for which both TSH and fT(4) concentrations were measured, 18 (0.23%) had fT(4) and TSH exceeding the investigated limits. Of these, two were due to heterophile antibody interference (with the TSH assay), one was proven to be due to random error, four could be explained by the patients' conditions, three were from infants <1 week of age, and the remaining eight were unresolved, primarily due to insufficient sample, discarded samples, and inability to obtain patient histories. CONCLUSIONS: This study defined a rules-based alert system for clinically unlikely combinations of TSH and fT(4) results. This pilot study demonstrates that this system is capable of detecting at least two different types of laboratory errors that would have otherwise gone undetected. PMID- 12763286 TI - Alterations of HDL subclasses in hyperlipidemia. AB - BACKGROUND: It is generally accepted that different high-density lipoprotein (HDL) subclasses have distinct but interrelated metabolic functions. HDL is known to directly influence the atherogenic process and changes in HDL subclasses distribution may be related to the incidence and prevalence of atherosclerosis. METHOD: The relative apolipoprotein (apo)A-I contents (% apoA-I) of plasma HDL subclasses were determined by two-dimensional gel electrophoresis coupled with immunodetection for apoA-I, in 39 hypercholesterolemic (HTC) subjects, 97 hypertriglyceridemic (HTG) subjects and 32 mixed hyperlipidemic (MHL) subjects, and 124 normolipidemic subjects. RESULTS: The relative apoA-I contents of prebeta(1)-HDL, prebeta(2)-HDL, HDL(3c), HDL(3b) and HDL(3a) significantly increased while HDL(2a) and HDL(2b) significantly decreased in hyperlipidemic subjects. In HTC subjects of hyperlipidemia, the concentrations of prebeta(1)-HDL were significantly lower and HDL(2b) concentrations were significantly higher than in HTG and MHL subjects. In HTG subjects, the concentrations of HDL(3a) were significantly higher and the concentrations of HDL(2b) were lower than in HTC and MHL subjects. In total hyperlipidemic subjects, plasma triglyceride (TG) concentrations showed positive correlation with prebeta(1)-HDL, prebeta(2)-HDL, HDL(3b) and HDL(3a) and negative correlation with HDL(2a) and HDL(2b). The total cholesterol (TC) concentrations showed positive correlation with the relative apoA-I contents of prebeta(1)-HDL and HDL(3b), whereas the HDL-C concentrations showed negative correlation with the relative apoA-I contents of prebeta(1)-HDL and HDL(3a) and positive correlation with those of HDL(2a) and HDL(2b). The relative apoA-I contents of prebeta(1)-HDL, prebeta(2)-HDL, HDL(3b), and HDL(3a) were positively correlated whereas those of HDL(2a) and HDL(2b) were negatively correlated with TG/HDL-C ratio. CONCLUSION: The particle size of HDL in hyperlipidemic subjects shifted towards smaller sizes, which, in turn, indicates that the maturation of HDL may be abnormal in hyperlipidemic subjects. PMID- 12763287 TI - Transferrin microheterogeneity in pregnancies with preeclampsia. AB - BACKGROUND: It has been reported that concentrations of serum transferrin (Tf) and its highly sialylated subfraction increase in normal pregnancy. This study investigated changes in the concentrations of serum transferrin and its subfractions in preeclampsia. METHODS: The serum concentration of transferrin was determined by a standard turbidimetric assay and microheterogeneous transferrin subgroups (low sialylated, 4-sialo and highly sialylated transferrins) were assessed by crossed immuno-isoelectric focusing. RESULTS: Compared to normal pregnancy, the concentrations of total, 4-sialo and highly sialylated transferrins decreased by 27%, 16% and 38%, respectively, in severe preeclampsia, while these values did not significantly decrease in mild preeclampsia. The concentration of low sialylated transferrin involving 2-sialo- and 3-sialo transferrins significantly decreased both in mild and severe preeclampsia, the value in severe preeclampsia was even significantly lower than that in nonpregnant women. The serum concentrations of total and highly sialylated transferrins in preeclampsia were correlated positively with infant birth weights (r=0.587 and r=0.645, respectively). CONCLUSIONS: The serum concentrations of total and highly sialylated transferrins in severe preeclampsia decrease significantly. This might have a negative impact on intrauterine growth. Additionally, the serum low sialylated transferrin decreases more sensitively in preeclampsia, although the concentration is low even in normal pregnancy. PMID- 12763289 TI - Plasma coenzyme Q10 reference intervals, but not redox status, are affected by gender and race in self-reported healthy adults. AB - BACKGROUND: Abnormal concentrations of coenzyme Q(10) have been reported in many patient groups, including certain cardiovascular, neurological, hematological, neoplastic, renal, and metabolic diseases. However, controls in these studies are often limited in number, poorly screened, and inadequately evaluated statistically. The purpose of this study is to determine the reference intervals of plasma concentrations of ubiquinone-10, ubiquinol-10, and total coenzyme Q(10) for self-reported healthy adults. METHODS: Adults (n=148), who were participants in the Princeton Prevalence Follow-up Study, were identified as healthy by questionnaire. Lipid profiles, ubiquinone-10, ubiquinol-10, and total coenzyme Q(10) concentrations were measured in plasma. The method used to determine the reference intervals is a procedure incorporating outlier detection followed by robust point estimates of the appropriate quantiles. RESULTS: Significant differences between males and females were present for ubiquinol-10 and total coenzyme Q(10). Blacks had significantly higher Q(10) measures than whites in all cases except for the ubiquinol-10/total Q(10) fraction. CONCLUSIONS: The fraction of ubiquinol-10/total coenzyme Q(10) is a tightly regulated measure in self reported healthy adults, and is independent of sex and racial differences. Different reference intervals for certain coenzyme Q(10) measures may need to be established based upon sex and racial characteristics. PMID- 12763288 TI - Alteration in the glutathione, glutathione peroxidase, superoxide dismutase and lipid peroxidation by ascorbic acid in the skin of mice exposed to fractionated gamma radiation. AB - BACKGROUND: In spite of the immense therapeutic gains produced by the fractionated irradiation (IR) regimen, radiation burden on the skin increases significantly. Protection of skin might enable use of higher radiation doses for better therapeutic gains. Ascorbic acid (AA), an essential ingredient of the human diet, is known to be a free radical scavenger and radioprotective agent. This study was undertaken to evaluate the effect of ascorbic acid on the radiation-induced changes in the status of glutathione (GSH), glutathione peroxidase (GPx), superoxide dismutase (SOD) and lipid peroxidation (LPx) in the skin of mice exposed to 10, 16 and 20 Gy of fractionated gamma radiation. METHODS: One group of the animals was administered daily with double distilled water (DDW), while the other group received 250 mg/kg b. wt. of ascorbic acid once daily, consecutively for 5, 8 or 10 days, before hemibody (below rib cage) exposure to 2 Gy/day of gamma-rays. Skin biopsies from both the groups were collected for the biochemical estimations. RESULTS: The irradiation of animals resulted in a dose-dependent decline in the activities of superoxide dismutase, glutathione peroxidase and glutathione contents. Ascorbic acid pretreatment resulted in a significant increase in the activities of both the enzymes and glutathione in the irradiated mouse skin. Normal concentrations of glutathione could not be restored even by day 6 post-irradiation. Conversely, lipid peroxidation increased in a dose-dependent manner in both the groups reaching a peak concentration by 3 h post-irradiation, while the ascorbic acid pretreatment inhibited the radiation-induced increase in lipid peroxidation. CONCLUSIONS: The ascorbic acid treatment arrested the decline in the activities of superoxide dismutase and glutathione peroxidase, glutathione contents and inhibited the radiation-induced lipid peroxidation in the skin of mice exposed to different doses of fractionated gamma radiation. PMID- 12763290 TI - Major depression and heat shock protein 70-1 gene. AB - BACKGROUND: Heat shock protein (HSP) expression can be induced by any stress such as with adrenocorticotropic hormones and catecholamines. It has been reported that patients with major depression have a 162-base deletion in the 5'-flanking region of heat shock protein 70 (HSP70)-1 gene mRNA. METHODS: To detect the HSP70 1 gene mRNA, total RNA was isolated and amplified by RT-PCR, and the sequence was confirmed in all five patients by DNA direct sequencing analysis. RESULTS: RT-PCR produced was no deletions of 162 bp in the human heat shock protein 70-1 gene in any of the patients with major depression or the nine controls. CONCLUSION: This finding is inconsistent with previous reports. We suggest that the 162-base deletion in the 5'-flanking region of the HSP70-1 gene mRNA is not associated with major depression. Further studies are required to determine the amounts of HSP70 and its mRNA in stress disorders such as major depression. PMID- 12763291 TI - Rheumatoid factor and false positive sex-hormone binding globulin. PMID- 12763292 TI - Altered amino acid levels in sera of a mouse model for juvenile neuronal ceroid lipofuscinoses. PMID- 12763293 TI - Baseline personality functioning correlates with 6 month outcome in schizophrenia. AB - OBJECTIVE: The assessment of outcome in schizophrenic patients should consider both the response to treatment and the recovery of social skills. The aim was to evaluate the outcome and related psychostructural and clinical factors in schizophrenic patients after they underwent 6 months of residential multimodal treatment. METHOD: Fifty-two schizophrenic patients enrolled in a multimodal treatment program were included in the study. Symptomatology and social functioning were assessed with the Brief Psychiatric Rating Scale (BPRS) and the Social and Occupational Functioning Assessment Scale (SOFAS). The Karolinska Psychodynamic Profile (KAPP) was used for the psychostructural evaluation. RESULTS: After 6 months there was a significant improvement in the global scores of BPRS, SOFAS, and some areas of KAPP. The personality (KAPP) and social occupational functioning (SOFAS) at baseline (T0) correlated with the global score of BPRS at 6 months (T6); moreover, SOFAS at T6 correlated with BPRS and KAPP at T0 and with the illness duration. CONCLUSION: The better the personality functioning in schizophrenic patients the better seems to be the response to treatment, with regard to symptoms as well as rehabilitation. Personality assessment might be useful for the individualisation of therapies, even within the context of a standardised program. PMID- 12763294 TI - Oral health and treatment needs of institutionalized chronic psychiatric patients in Israel. AB - The aim of the study was to examine the oral health and treatment needs of chronically hospitalized psychiatric patients in Israel. Ten percent of the patients hospitalized for more than 2 years in the 18 psychiatric institutions in Israel were selected at random. The dental status (DMF-T index) was calculated, demographic and medical data were retrieved from the files. Of the 431 patients examined (250 men, 181 women, average age 54 years) 312 patients had only partial natural dentition. The average DMF-T score was 26.74 (out of 32), one of the highest in the literature. The caries component accounted for 2.3% of the DMF-T, the missing teeth component 72% and the restored teeth component 5%. There was an adverse correlation between age and caries and between duration of hospitalization and number of teeth. The average number of carious and missing teeth was higher than in the healthy population. No all-edentulous patients had dentures. These findings confirm the urgent need for an intervention program to improve dental health care in high-risk, difficult-to-treat, psychiatric chronic inpatients. PMID- 12763295 TI - Normal and obsessional jealousy: a study of a population of young adults. AB - BACKGROUND: Jealousy is a heterogenous emotion ranging from normality to pathology. Several problems still exist in the distinction between normal and pathological jealousy. AIM OF THE STUDY: With the present study, we aimed to contribute to the definition of the boundary between obsessional and normal jealousy by means of a specific self-report questionnaire developed by us. METHOD: The questionnaire called "Questionnaire on the Affective Relationships" (QAR) and consisting of 30 items, was administered to 400 university students of both sexes and to 14 outpatients affected by obsessive-compulsive disorder (OCD) whose main obsession was jealousy. The total scores and single items were analysed and compared. RESULTS: Two hundred and forty-five, approximately 61% of the questionnaires, were returned. The statistical analyses showed that patients with OCD had higher total scores than healthy subjects; in addition, it was possible to identify an intermediate group of subjects, corresponding to 10% of the total, who were concerned by jealousy thoughts around the partner, but at a lower degree than patients, and that we called "healthy jealous subjects" because they had no other psychopathological trait. Significant differences were also observed for single items in the three groups. CONCLUSIONS: Our study showed that 10% of a population of university students, albeit normal, have jealousy thoughts around the partner, as emerged by the specific questionnaire developed by us. This instrument permitted to clearly distinguish these subjects from patients with OCD and healthy subjects with no jealousy concern. PMID- 12763296 TI - Late onset alcoholism. AB - Rather high prevalence rates of alcohol abuse in the elderly have been reported in the literature. However, there is some evidence that many elderly persons with alcohol problems are not identified, probably due to the nonspecificity of alcohol-related presentations in old individuals. Thus, there is an ongoing discussion on appropriate diagnostic criteria for alcohol dependence in elder people who frequently begin to abuse alcohol in late life. This study was aimed to explore whether alcoholics with late onset (beginning after the age of 45) differ from those with an early onset (prior the age of 25). Two hundred and sixty eight subjects consecutively referred to a ward of a general hospital specialized for alcohol detoxification were divided into three groups by the age at onset of harmful alcohol consumption. The duration of harmful drinking was rather similar in all groups. However, alcohol dependence according to the ICD-10 criteria (three or more have to be fulfilled) was diagnosed in 94.1% of the alcoholics with an early onset (/= 16); 77 patients having a less severe depression (BDI < 16); 32 patients with mild or non-existent depression (BDI 14 days). Validity of the water swallow test as a predictor of aspiration pneumonia was confirmed. Significant associations for prolonged dysphagia were seen with stroke severity, dysphasia and lesions of the frontal and insular cortex on brain imaging. These results indicate that it may be possible to predict patients who will develop prolonged significant dysphagia following acute stroke thereby facilitating referral for insertion of PEG at an earlier time point. PMID- 12763333 TI - Blink reflex alterations in recently diagnosed diabetic patients. AB - OBJECTIVE: The aim of the present study was to determine the frequency of blink reflex alterations and to examine the influence of hyperglycemia in inducing the alterations in recently diagnosed Type 2 diabetes mellitus patients. METHODS: A cross-sectional study was carried out on patients having asymptomatic diabetes with a period of evolution under 10 years. In all 47 patients (26 women and 21 men), serum glycemia levels were determined and the latency onset of the blink reflex components were measured. RESULTS: The average patient age was 44.5+/-11.0 (mean+/-SD) years with a diabetes evolution period of 4.3+/-2.9 (mean+/-SD) years. After a fasting serum glucose test, the diabetic patients were catalogued as normoglycemic (< or =126 mg/dl) or as hyperglycemic (> 26 mg/dl) and subjected to a blink reflex test. The results obtained from the diabetic patients were compared with those from a non-diabetic control group. 14.8-31.9% of the diabetic patients showed alterations in blink reflex component latencies. The differences compared with the control group were significant (p<0.05). CONCLUSIONS: Diabetes, as is well-known, can affect the central and peripheral nervous system and there does not appear to be a link between glycemic levels and blink reflex components. However, blink reflex alterations were present even in diabetic patients with a relatively short period of disease evolution. PMID- 12763334 TI - Deep cerebral venous sinus thrombosis often presents with neuropsychologic symptoms. AB - The outcome of cerebral venous sinus thrombosis (CVST) depends on rapid diagnosis and initiation of effective anticoagulation. We report seven cases of a subgroup with deep cerebral venous sinus thrombosis (DCVST) treated in our institution since 1990. Six of our seven patients presented with early neuropsychological deficits (mental obtundation, bradyphrenia or apathia). This clinical presentation, in combination with headache, and focal neurological deficits, aids the early diagnosis of DCVST. Thalamic hyperintensities on T2-weighted MRI images, previously considered infarctions, were fully reversible during treatment with heparin. This indicates that early in the course of the disease they correspond to vasogenic oedema. PMID- 12763335 TI - Genetic and environmental risk factors and their interactions for Parkinson's disease in a Chinese population. AB - The interaction between genetic and environmental factors for PD was examined in a Chinese population. It was found that although the intron 2 MAOB (GT)(n) repeat polymorphism was not associated with PD in the population, a relationship might have been masked by the "protective effect" of tea drinking. In individuals who did not drink tea (<1 cup/day), the possession of short length < or = 178 bp (GT)(n) alleles conferred a borderline significant increased risk for PD (adjusted OR=1.47; C.I.=1.03-2.1). As the extent of tea consumption increased, the association between the < or = 178 bp allele and PD disappeared. This result suggests that the MAOB gene may be associated with PD in Chinese if the putative protective effect of tea drinking is taken into account. The significance of this finding is unclear as the study may be limited because of its marginal significance and limited numbers. However, it does demonstrate the importance of considering putative positive and negative environmental risk factors in any examination of genetic risk factors for PD. PMID- 12763336 TI - Differences in acetazolamide vasoreactivity in patients with acute and chronic occlusion of the internal carotid artery. AB - Using a xenon-enhanced computed tomography (Xe-CT) system, we measured cerebral blood flow (CBF) and acetazolamide vasoreactivity in 3 major vascular territories, the thalamus and putamen on both sides in 3 groups of patients undergoing the balloon occlusion test (BOT), with internal carotid occlusion (ICO) and with open carotids.In the ICO group, although the CBFs on the occluded side were similar to those in the BOT group, the vasoreactivity was significantly higher. The CBFs on the contralateral side were lower than those in the BOT group, but the vasoreactivity was similar. The vasoreactivity of the MCA on the occluded side mostly correlated with that in the other areas, similar to the results obtained in the open carotid group, although it was poorly correlated with other areas in the BOT group. Carotid artery occlusion may reduce CBF in a large area with restoration of vasoreactivity in the chronic stage. PMID- 12763337 TI - The association of the expression of vascular endothelial growth factor with the cystic component and haemorrhage in pituitary adenoma. AB - Vascular endothelial growth factor (VEGF) is known to be a mediator of angiogenesis and vascular permeability. A cystic component and haemorrhage are often found in pituitary adenomas. We assessed the VEGF expression based on immunohistochemical examinations in 48 pituitary adenomas. All the adenomas showed some VEGF immunoreactivity mainly in the cytoplasm of tumour cells. Of the 48 adenoma-cases, 16 cases had a strong VEGF immunoreactivity, 26 cases had a moderate one, and 6 cases had a weak one. On the MR images, a cystic component was found in 16 cases (33.3%), and a haemorrhage was found in 18 cases (37.5%). The VEGF immunoreactivity had a significant relationship with the cystic component but not the haemorrhage, size, recurrence, or HE classification. These findings suggest that VEGF plays any potential role in the pathogenesis of cystic formation in pituitary adenomas. PMID- 12763338 TI - Rhabdoid glioblastoma: a case report. AB - The authors report a 16 year old girl with a supratentorial rhabdoid glioblastoma. The radiological features, histopathology and management of this rare variant of glioblastoma multiforme are discussed. PMID- 12763339 TI - Effect of magnesium sulphate in experimental spinal cord injury: evaluation with ultrastructural findings and early clinical results. AB - Excitotoxic mechanisms have been implicated in the pathophysiology of spinal cord injury (SCI). The authors have studied the protection against secondary damage to rat spinal cord with magnesium sulphate, a well-known N-methyl-D-aspartate antagonist. Rats were randomly allocated into 5 groups. Group 1 rats were controls and normal spinal cord samples were obtained after clinical examination. 50 g-cm contusion injury was introduced to Group 2. Group 3 was vehicle, 1 cc of physiologic saline was injected post-trauma. Group 4 and 5 were treatment groups and 100 mg/kg and 600 mg/kg of Magnesium sulphate was given immediately after trauma, intraperitoneally. Animals were evaluated with inclined plane, Tarlov motor scale and Basso-Beattie-Bresnahan scale 24h after SCI. Spinal cord samples for ultrastructural evaluations were obtained following clinical examinations. Magnesium treatment improved neurological outcome. Electron microscopic results showed obvious neuroprotection in the treatment groups. Application of 600 mg/kg of magnesium revealed better ultrastructural findings and clinical results than 100 mg/kg. These findings demonstrated that magnesium sulphate possesses neuroprotection on spinal cord ultrastructure and on functional scores after acute contusion injury to the rat spinal cord. PMID- 12763340 TI - Proliferating cell populations in experimentally-induced hydrocephalus in developing rats. AB - To examine the fate of proliferating brain cells in hydrocephalus (Hydro), experimental Hydro was induced in neonatal rats by intracisternal injection of kaolin and, 3 weeks later, the rats were injected with bromodeoxyuridine (BrdU). The BrdU (+) cells were immunohistochemically analyzed by using antibodies against neural (nestin), neuronal (NeuN) and glial (GFAP and MBP) markers in the posterior cerebrum. The percentage of nestin expression for the BrdU (+) cells was 8% in control and increased from 17% in the Hydro to 33% in the Hydro at an earlier stage after the shunt procedure, but was restored to 6% in the Hydro at a later stage after the shunt procedure. The percentages of GFAP expression showed a similar tendency to those of nestin expression. The BrdU (+) cells did not express either NeuN or MBP throughout the experiments. PMID- 12763341 TI - Treatment of painful peripheral neuroma by vein implantation. AB - Painful neuromas form on cutaneous nerves as a result of trauma, pressure, stretch or entrapment. Since the earliest descriptions of neuromas, proposed treatments have been met with poor results and controversy. The myriad of treatments described include: simple division of an affected nerve, implantation into muscle or bone, silicon sleeves and caps, repeated injection of steroids, end-to-side neurorrhaphy, medication and vein caps to name a few. Due to encouraging recent reports of treatment of painful neuromas by vein implantation, the authors describe a simple technique to achieve this surgical goal. As veins are readily accessible due to their proximity in the neurovascular bundle, they serve as a ready source for grafting. The advantages include minimisation of trauma to bone and muscle as compared with previous treatment techniques and the relative ease of the method. PMID- 12763342 TI - Selected midfacial access procedures to the skull base. AB - The indications and operative technique of various procedures commonly used to provide or increase access to the central skull base, anterior and middle cranial fossae, nasopharynx, infratemporal fossa and retromaxillary space are discussed with illustrative cases. PMID- 12763343 TI - Angiolipoma of the right inferior colliculus: a rare central cause of hearing loss and limb ataxia. AB - Intracranial angiolipomas are rare entities and are infrequently symptomatic. These benign lesions are adherent to the surrounding neurovascular structures as they share the same blood supply, which limits their resectability, and impose a possibility of persistence of symptoms. However, due to recent advancements in the neuro-radiological fields and microneurosurgical techniques it has been possible to localise precisely and excise these lesions with low morbidity and mortality. The authors present an interesting rare case of intracranial angiolipoma of the right inferior colliculus situated in the quadrigeminal plate cistern. The patient had presented with ipsilateral hearing loss and upper limb ataxia. Interesting clinical findings, neuro-imaging studies and peroperative features are presented and discussed along with a brief review of the literature. PMID- 12763344 TI - Giant cerebral cavernous haemangioma: a case report and review of literature. AB - An 18-year-old male presented with uncontrolled left focal seizures with secondary generalisation for the past 10 years. Investigations revealed a large lobulated mass in the right frontal brain. Surgical excision of a giant cavernous haemangioma was performed. The patient is seizure-free following the surgery. The case and relevant literature on the rare entity of giant intracranial cavernous haemangiomas is discussed. PMID- 12763345 TI - Fatal brain stem event complicating acute pancreatitis. AB - Acute pancreatitis, developing in a patient with chronic renal failure, was complicated by a fatal neurological illness during which MRI showed pontine and extrapontine changes consistent with pontine and extrapontine myelinolysis. At post mortem, acute pancreatitis was confirmed but the neuropathological findings were more in keeping with an unusual presentation of acute haemorrhagic leucoencephalitis, perhaps even representing a form of 'pancreatic encephalopathy'. Although the development of CT and MRI scanning has greatly increased the resolution of neuroimaging and facilitated diagnosis during life, the value of autopsy examination is confirmed in cases such as this. Sometimes the findings may raise more questions than may have been answered - this too is an important function! PMID- 12763346 TI - A case of mucopolysaccharidosis IV with lower leg paresis due to thoraco-lumbar kyphoscoliosis. AB - We treated a patient of type IV mucopolysaccharidosis (Morquio's disease) with lower leg paresis due to kyphoscoliosis. A 65-year-old woman presented with Morquio's disease. A lateral radiograph demonstrated the classic bullet-shaped vertebrae and a 65 degrees thoraco-lumbar kyphosis. After the age of 60, she suffered from numbness in both lower legs and walking disturbance. Bilateral patellae-tendon reflexes were exaggerated. MRI showed compression of the spinal cord around T12 to L2 with a highlighted area of change inside the spinal cord. Myelography and computed tomography after the myelography showed narrowing of the sub-arachnoidal space and deformation of the spinal cord around the T12 to L2 levels. Severe vertebral osteoporosis made it necessary to first perform posterior correction of the kyphosis and fusion. The curve was stabilised with the Luque method from T7 to L4. Her neurological condition markedly recovered, but 1 year after surgery her neurological condition again began to deteriorate, resulting in walking disturbance. For this reason, anterior decompression and fusion through a lateral thoracotomy was undertaken. Decompression of the spinal cord and a bone graft from the iliac crest were attained. The patient's neurological condition again improved, but not as much as immediately after the first operation. PMID- 12763347 TI - Surgical management of massive cerebellar infarction. AB - Massive cerebellar infarction accounts for less than 2% of strokes. Unlike massive hemispherical infarctions, in pure cerebellar infarctions, the prognosis is better. This case report discusses, a 61 year old lady who presented with atrial fibrillation and a massive cerebellar infarction. Timely surgical intervention reversed the deterioration in neurological status. The indications for surgical management, based on review of the literature, is presented. PMID- 12763348 TI - Large cystic fibrous dysplasia of the temporal bone: case report and review of literature. AB - Fibrous dysplasia is an uncommon benign disorder of unknown aetiology. It most likely represents a disorder of normal bone development. As it may occasionally involve the temporal bone, and encroach on the middle cranial fossa, it is of interest to the neurosurgeon. Fibrous dysplasia of the cystic variety is very uncommon. We report a case of a large cystic fibrous dysplasia involving predominantly the squamous temporal bone. The radiology and histology are discussed, and its differentiation from other lesions. PMID- 12763349 TI - Dyke-Davidoff-Masson syndrome manifested by seizure in late childhood: a case report. AB - The patient was a 19-year-old woman who presented with hemiatrophy and diminished superficial sensation on the left side of her body including her face. She had a past history of tonic-clonic seizures accompanied by left hemiparesis in late childhood. Brain CT demonstrated dilatation of the frontal sinus, calvarial thickening, cerebral hemiatrophy and dilatation of the lateral ventricle on the right side. Brain MRI showed atrophy of the right cerebrum and midbrain and dilatation of the lateral ventricle on T1-weighted images, as well as a high signal intensity area from the parietal to the occipital lobe on T2-weighted images. These findings are suggestive of an episode that may have caused a transient ischemia through the right cerebral hemisphere after the intrauterine period. PMID- 12763350 TI - Theta oscillation in the anterior cingulate and beta-1 oscillation in the medial temporal cortices: a human case report. AB - In previous studies we observed gamma (30-150 Hz) and beta-1 (10-20 Hz) oscillations in the medial temporal lobe (MTL) using subdural electrodes. The beta-1 was present during wake and REM sleep while gamma was present in all states. Recently we studied a patient (35 years M) with electrodes attached to the anterior cingulate cortex (ACC). This structure showed regular theta (5-6 Hz) oscillations. In the first recording, electrodes were attached to the MTL, that demonstrated the beta-1 and gamma oscillations. Two months later, electrodes were placed on orbitofrontal and ACC and an all night sleep recording was carried out. The ACC exhibited a highly regular and continuous theta oscillation during wakefulness and REM sleep, but not during NREM sleep. Since this same subject showed beta-1 oscillations in the MTL, it is probable that the theta in the ACC is independent of beta-1 in the MTL. This single case suggests the existence of two different frequency oscillators in the human limbic system. Elucidating their functional roles will be an interesting challenge for future studies. PMID- 12763351 TI - Entrapment neuropathy of the ulnar nerve by a constriction band: the role of MRI. AB - The diagnosis of ulnar nerve entrapment at the elbow has relied primarily on clinical and electrodiagnostic findings. Magnetic resonance imaging (MRI) has been used in the evaluation of peripheral nerve entrapment disorders to document signal and configurational changes in nerves. In this case report we review the MRI and operative findings of a rare constriction band causing ulnar nerve compression at the elbow. We review the sensitivity and specificity in diagnosing ulnar nerve entrapment at the elbow as defined by MRI findings. PMID- 12763352 TI - Improvement in cognitive function after radical excision of an anterior skull base meningioma--a report of 2 cases. AB - It has been well documented that memory difficulties in patients with anterior skull base meningioma may improve after tumour resection. However, there have been few reports on studies where precise testing of cognitive function has been conducted before and after surgery. Here we report 2 cases of anterior skull base meningioma where 2 tests of cognitive function, the Wechsler adult intelligence scale-revised (WAIS-R) and the Wechsler memory scale-revised (WMS-R), were performed by the patient before and after surgical treatment. After the operation there was a dramatic increase in the performance IQ as measured by the WAIS-R. In addition, the scores for both the "Attention/Concentration" and the "Delayed Recall" subsets of the WMS-R scale improved. There were differences between the 2 patients in the length of time required for functional recovery and this was thought to be due to the tumour size and the age of the patient. The use of these scales would enable physicians who are treating patients with anterior skull base meningioma to assess the recovery time that is likely to be needed before full return of cognitive function after surgery. PMID- 12763353 TI - Cerebellar haemorrhage following supratentorial craniotomy. AB - Distant cerebellar haemorrhage is a rare complication occurring in approximately 0.3-0.6% of all supratentorial craniotomy. A Medline and Pubmed search revealed only 98 cases in the English literature. We report three cases from our institution. An overall review of these 101 cases demonstrated that this complication commonly presented early in the postoperative course as decreased level of consciousness following aneurysm repair surgery or lobectomy for epilepsy. Asymptomatic presentation due to small haemorrhage was not uncommon. A transtentorial pressure gradient set up by excessive CSF loss is generally held responsible for disrupting the cerebellar venous blood flow and consequently leading to venous haemorrhage. Perioperative hypertension may also play a role. The outcome of patients who survived the complications was generally good, though not infrequently fatality resulted from the mass effect of extensive cerebellar haemorrhage demanded vigilance in its management. PMID- 12763354 TI - Pathological laughter as a presenting symptom of acoustic schwannoma: report of two cases. AB - Two cases of acoustic schwannoma in a 48-year-old woman and 18-year-old male are reported. Both patients presented with an extremely unusual symptom of pathological laughter as a principal presenting symptom. Complete resection of the tumour via a retrosigmoid route resulted in an immediate resolution of pathological laughter. PMID- 12763355 TI - Sensorimotor polyneuropathy with 5-aminosalicylic acid: a case report. AB - A 69-year-old man with ulcerative colitis (UC) developed sensorimotor polyneuropathy. First, he received salazosulphapyridine (SASP) as treatment for the UC. The symptoms of UC disappeared immediately, but he developed skin eruptions and dysesthesia in his lower limbs. When SASP was changed to 5 aminosalicylic acid (5-ASA), his skin eruptions were resolved, however, he developed weakness and atrophy in his right arm as well as progressive worsening of the dysesthesia in his legs and gait disturbance. Deep tendon reflexes (DTR) were absent in all extremities. After 5-ASA was discontinued, the polyneuropathy symptoms recovered gradually. This clinical course suggests that the sensorimotor polyneuropathy may have been caused by 5-ASA. PMID- 12763357 TI - Multiple schwannomas of the sciatic nerve. AB - Schwannomas are rare benign tumours of nerve sheath cells of neural crest origin. Often these tumours are solitary and encapsulated. Multiple schwannomas can arise from the peripheral nervous system including cranial nerves, spinal roots, the brachial and lumbar-sacral plexus or major peripheral nerves. We report an extremely rare case of schwannomatosis of the sciatic nerve in a young female and include a comprehensive literature review. Treatment options are discussed. PMID- 12763356 TI - Cervical ependymoma presenting with brainstem and cerebellar signs: case report. AB - This case report demonstrates cervical spinal cord pathology which presented with brainstem and cerebellar signs consequent to the peritumoural oedema that extended rostrally to the pontomedullary junction. A Medline search of the literature back to 1960 failed to produce any previous report of a cervical ependymoma presenting with brainstem and cerebellar signs purely consequent to oedema. This case highlights the need to look further afield when presented with the scenario of clinical features of a brainstem lesion with only oedema apparent on cranial imaging. It indicates the need to include cervical imaging well below the foramen magnum in these circumstances. PMID- 12763358 TI - Re: Basilar invagination and Chiari malformation associated with cerebellar atrophy: report of two treated cases. PMID- 12763359 TI - Re: Magnesium: a useful adjunct in the prevention of cerebral vasospasm following aneurysmal subarachnoid haemorrhage. PMID- 12763361 TI - The discovery of the new haemochromatosis gene. 1996. PMID- 12763362 TI - Relation between hepatic expression of ATP-binding cassette transporters G5 and G8 and biliary cholesterol secretion in mice. AB - BACKGROUND/AIM: Mutations in genes encoding the ATP-binding cassette (ABC) transporters ABCG5 and ABCG8 underlie sitosterolemia, which is characterized by elevated plasma levels of phytosterols due to increased intestinal absorption and impaired biliary secretion of sterols. The aim of our study was to correlate the expression levels of Abcg5 and Abcg8 to biliary cholesterol secretion in various (genetically-modified) mouse models. METHODS: Bile was collected from genetically modified mice fed a chow diet, or from mice fed either a chow diet, or chow supplemented with either 1% diosgenin, 0.1% simvastatin, or a synthetic liver X receptor agonist, for determination of biliary lipids. Livers and small intestines were harvested and expression levels of Abcg5, Abcg8 and Abcb4 were determined by real-time polymerase chain reaction. RESULTS: Intestinal expression of Abcg5 and Abcg8 did not show much variation between the various models. In contrast, a linear correlation between hepatic expression levels of Abcg5 and Abcg8 and biliary cholesterol secretion rates was found. This relation was independent of Abcb4-mediated phospholipid secretion. However, in diosgenin-fed mice showing cholesterol hypersecretion, hepatic Abcg5 and Abcg8 expression levels remained unchanged. CONCLUSIONS: Our results strongly support a role for Abcg5 and Abcg8 in regulation of biliary cholesterol secretion, but also indicate the existence of a largely independent route of cholesterol secretion. PMID- 12763363 TI - Adaptive changes in hepatobiliary transporter expression in primary biliary cirrhosis. AB - BACKGROUND/AIMS: Information about alterations of hepatobiliary transporter expression in primary biliary cirrhosis (PBC) could provide important insights into the pathogenesis of cholestasis. This study aimed to determine the expression of hepatobiliary transport systems for bile salts (Na(+)/taurocholate cotransporter, NTCP; bile salt export pump, BSEP), organic anions (organic anion transporting protein, OATP2; canalicular conjugate export pump, MRP2; basolateral MRP homologue, MRP3), organic cations (canalicular multidrug export pump, MDR1), and phospholipids (canalicular phospholipid flippase MDR3) in livers from patients with advanced stages of PBC. METHODS: Transporter mRNA and protein levels were assessed by reverse transcription polymerase chain reaction and Western blot analysis. Tissue distribution of transporters was investigated by immunohistochemistry and immunofluorescence microscopy. Hepatic bile acids were measured by gas chromatography-mass spectrometry. RESULTS: Compared to controls, basolateral uptake systems (NTCP, OATP2) were reduced, canalicular export pumps for bile salts and bilirubin (BSEP, MRP2) were preserved, while canalicular MDR P glycoproteins (MDR1, MDR3) and the basolateral efflux pump MRP3 were increased in PBC. Double immunofluorescence labeling with a canalicular marker (dipeptidyl peptidase IV) demonstrated proper canalicular localization of BSEP and MRP2 in PBC. OATP2 and MRP2 expression correlated inversely with hepatic levels of hydrophobic bile acids, while positively correlating with hepatic enrichment with ursodeoxycholic acid. CONCLUSIONS: Down-regulation of basolateral uptake systems and maintenance/up-regulation of canalicular and basolateral efflux pumps may represent adaptive mechanisms limiting the accumulation of toxic biliary constituents. PMID- 12763364 TI - Sensitization by interleukin-6 of rat hepatocytes to tumor necrosis factor alpha induced apoptosis. AB - BACKGROUND/AIMS: Tumor necrosis factor (TNF) elicits hepatocyte apoptosis in toxic liver injury and is also central in hepatocyte proliferation after partial hepatectomy. In both circumstances interleukin (IL)-6 levels are also elevated. In mouse liver IL-6 attenuated Fas receptor-mediated apoptosis indicating its interference with pro-apoptotic signal chains. It was, therefore, the aim to examine the modulation by IL-6 of TNFalpha-induced apoptosis in rat hepatocytes. METHODS: Primary rat hepatocytes were treated with IL-6 prior to induction of apoptosis with TNFalpha/actinomycin D or anti-Fas antibody M-20. Apoptosis was detected by determination of caspase-3 activation and bisbenzimide staining of condensed nuclei. Expression of TNFalpha receptors was analyzed by semi quantitative polymerase chain reaction and ligand binding studies with [125I] TNFalpha. RESULTS: IL-6 treatment doubled TNFalpha/actinomycin D-induced caspase 3 activity and significantly enhanced chromatin condensation. By contrast IL-6 inhibited Fas-induced increase in caspase-3 activity by 45% and significantly reduced chromatin condensation. IL-6 increased the mRNA level of TNF-R1 1.35-fold and augmented cell surface binding of [125I]-TNFalpha 3-fold. The latter and TNFalpha-mediated caspase activation was attenuated by prostaglandin E(2). CONCLUSIONS: IL-6 - in contrast to its anti-apoptotic modulation of the Fas induced pathway - exerted a pro-apoptotic effect on the TNFalpha/actinomycin D induced apoptosis by increasing the number of TNF-R on hepatocytes. PMID- 12763365 TI - Primary human hepatocytes--a valuable tool for investigation of apoptosis and hepatitis B virus infection. AB - BACKGROUND/AIMS: Apoptosis is a key event in the pathophysiology of many liver diseases. Primary human hepatocytes (PHH) provide a useful model to study physiological and pathophysiological processes in the liver. Our aim was to optimize PHH cultures to allow studies on induction of apoptosis and of hepatitis B virus (HBV) infection. METHODS: PHH were isolated from human liver tissue by two-step collagenase perfusion. PHH and hepatoma cells were treated with different apoptosis-inducing agents in parallel. PHH cultures were infected with wild type HBV and transduced with HBV genomes using adenoviral vectors. RESULTS: PHH were successfully isolated from 40 different tissue samples with high viability and purity. Perfusion time and seeding density turned out to be critical parameters for optimal cell yield and culture conditions, respectively. Serum addition to the medium reduced viability of PHH. PHH allowed reproducible studies of CD95-dependent and -independent apoptosis. Sensitivity towards CD95 mediated apoptosis was markedly higher than in hepatoma cells. PHH could efficiently be infected with HBV, but infection did neither induce apoptosis nor prevent CD95-induced cell death. CONCLUSIONS: Our data show that PHH provide an excellent tool for the investigation of apoptosis induced by agents like death receptor-ligands and hepatotropic viruses. PMID- 12763366 TI - Population screening for hemochromatosis: a study in 5370 Spanish blood donors. AB - BACKGROUND/AIMS: Hereditary hemochromatosis is associated with homozygosity for C282Y mutation in the HFE gene, elevated serum transferrin saturation and excess iron deposits throughout the body. We conducted a population-based study in Spain to asses the prevalence of the HFE mutations and their effect on iron parameters. METHODS: We screened 5370 blood donors for the C282Y and H63D HFE mutations by allele-specific polymerase chain reaction. Serum iron, serum ferritin and transferrin saturation were also measured. RESULTS: We have found eight (five men and three women) blood donors who are C282Y homozygotes (0.15%) and 74 C282Y/H63D compound heterozygotes (1.38%). Four out of the eight C282Y homozygotes, all men, had high serum ferritin and transferrin saturation values. No woman was detected with both iron parameters increased. Only one of the 74 C282Y/H63D compound heterozygotes showed elevated serum ferritin and transferrin saturation values (penetrance 1.35%). Serum ferritin and transferrin saturation were significantly higher in C282Y homozygous men as compared with the rest of the genotypes. CONCLUSIONS: The C282Y/C282Y genotype frequency in Spain is 1 in 1004. The C282Y/C282Y genotype is clearly associated with an increase in iron parameters. Biochemical expression of the disease was found in 80% of the C282Y/C282Y men. PMID- 12763367 TI - TIPS for acute and chronic Budd-Chiari syndrome: a single-centre experience. AB - BACKGROUND/AIMS: Transjugular intrahepatic portosystemic shunt (TIPS) is a technically challenging but feasible treatment for Budd-Chiari syndrome (BCS). However, information about the outcome, particularly in patients with liver failure, is scarce. We report our experience of TIPS for BCS. METHODS: Fifteen patients with BCS underwent TIPS. Eight had hepatic failure and seven underwent TIPS for BCS uncontrolled by medical treatment. RESULTS: Fourteen out of 15 had successful TIPS placement. Out of the eight hepatic failure patients, four died soon after TIPS: one liver rupture, one portal vein rupture, one liver failure and one pulmonary oedema. Another patient had a significant intrahepatic haematoma, which resolved with conservative management. TIPS was successfully placed in all of the seven patients with chronic BCS, in whom there was an average follow-up of 20 months. Ascites resolved and liver function improved in all. One patient died after 18 months from the original hepatic metastatic disease. Four patients have had evidence of TIPS dysfunction requiring three balloon dilatations and one restenting. No patient has required liver transplantation. CONCLUSIONS: TIPS should be the first line treatment for BCS uncontrolled by medical therapy. However, mortality in BCS with hepatic failure is high and liver transplantation could be a better option. PMID- 12763368 TI - Effect of the V1a/V2-AVP receptor antagonist, Conivaptan, on renal water metabolism and systemic hemodynamics in rats with cirrhosis and ascites. AB - BACKGROUND: Selective V(2)-AVP receptor antagonists are effective in inducing aquaresis in humans and rats with cirrhosis, hyponatremia and water retention. However, it is unknown whether dual V(1a)/V(2)-AVP antagonists are also efficacious as aquaretic agents under these conditions. This is important, particularly considering that blockade of V(1a)-AVP receptors could aggravate cardiocirculatory function in decompensated cirrhosis. AIMS: To evaluate the renal, hormonal and hemodynamic effects induced by the chronic oral administration of the V(1a)/V(2)-AVP antagonist, Conivaptan, in rats with CCl(4) induced cirrhosis, ascites and severe water retention. METHODS: We assessed the aquaretic efficacy of 10-day chronic oral administration of Conivaptan (0.5mg/kg body weight (bw)) in cirrhotic rats with hyponatremia and water retention. Urine volume (UV), osmolality (UOsm), and sodium excretion (U(Na)V) were measured daily. At the end of the study arterial pressure was also measured. RESULTS: Conivaptan produced an acute increase in UV, a reduction in UOsm and, at the end of the investigation, cirrhotic rats receiving the V(1a)/V(2)-AVP receptor antagonist did not show hyponatremia or hypoosmolality. Conivaptan also normalized U(Na)V without affecting creatinine clearance and arterial pressure. CONCLUSIONS: Dual V(1a)/V(2)-receptor antagonists may be therapeutically useful for the treatment of water retention and dilutional hyponatremia in human cirrhosis. PMID- 12763370 TI - Cytokeratin 7 staining of hepatocytes predicts progression to more severe fibrosis in alcohol-fed baboons. AB - BACKGROUND/AIMS: Not all alcoholic patients develop severe liver disease with fibrosis progressing to cirrhosis. It is of practical importance to determine whether some markers can predict progression of liver fibrosis. METHODS: We used a baboon model that mimics human alcoholic liver disease. Cytokeratin 7 and 19 expression and fat deposition were investigated in serial liver biopsies of 18 animals undergoing prolonged alcohol administration (range 2-17 years) and in four controls. Fibrosis was graded histologically and was also assessed quantitatively by image analysis. RESULTS: Ten animals did not show a progression of liver disease even after 17 years of alcohol administration, but eight animals fed alcohol exhibited a progression of liver disease from no fibrosis or perivenular fibrosis to septal fibrosis or cirrhosis within 7 years. In normal liver, cytokeratin 7 and cytokeratin 19 immunostaining is restricted to bile duct cells. Hepatocellular cytokeratin 7 was observed only in those animals which progressed to more severe stages of fibrosis and it anticipated this progression by 4.2 years on average. CONCLUSIONS: In alcohol-fed baboons, cytokeratin 7 staining of hepatocytes (but not cytokeratin 19, nor fat deposition) predicts with a high degree of sensitivity and specificity progression to more severe liver disease. PMID- 12763369 TI - Herbal medicine Inchin-ko-to (TJ-135) prevents liver fibrosis and enzyme-altered lesions in rat liver cirrhosis induced by a choline-deficient L-amino acid defined diet. AB - BACKGROUND/AIMS: The herbal medicine Inchin-ko-to (TJ-135), extract power from three herbs, has recently been reported possessing anti-apoptotic activity. The aim of this study was to investigate whether TJ-135 has any influence on the development of preneoplastic lesions as well as liver fibrosis. METHODS: The effects of the TJ-135 were examined using the choline-deficient L-amino acid defined diet-induced liver fibrosis model. In addition, the effect of TJ-135 on mitogen-activated protein (MAP) kinase, type III procollagen mRNA expression and the medium N-terminal procollagen III propeptide (PIIINP) concentration in a hepatic stellate cell line (LI90) were examined. RESULTS: TJ-135 prevented fibrosis in a dose-dependent manner up to 1.5% (w/w). TJ-135 also reduced the expression of type III procollagen mRNA in the liver, as well as the number of activated stellate cells. Furthermore, TJ-135 reduced the area of preneoplastic lesions in the liver. With LI90 cells, TJ-135 reduced MAP kinase (ERK and JNK but not P38) activities resulting in reduced type III procollagen mRNA and PIIINP concentrations in the medium in a dose-dependent manner. CONCLUSIONS: These results indicate that although TJ-135 has anti-apoptotic activity, TJ-135 does not increase preneoplastic lesions but significantly reduces liver fibrosis through the inhibition of stellate cell activation without a reduction of hepatocyte cell death. PMID- 12763371 TI - Induction of hepatic heme oxygenase-1 by diclofenac in rodents: role of oxidative stress and cytochrome P-450 activity. AB - BACKGROUND/AIMS: The role of oxidative stress in diclofenac hepatotoxicity is still not clear. This study examined whether the drug induced heme oxygenase-1 (HO-1), a stress protein. METHODS: HO-1 mRNA and HO activity were measured in mouse liver and in rat hepatocytes after treatment with diclofenac parallel to release of serum alanine aminotransferase (ALT) and sorbitol dehydrogenase (SDH) as a marker of hepatic damage. RESULTS: HO-1 was transcriptionally and dose dependently induced by diclofenac in mouse liver and rat hepatocytes. HO-1 mRNA, ALT and SDH peaked at the same time. Mechanistic studies revealed that the drug synergized with buthionine sulfoximine (BSO) in lowering hepatic glutathione, increased the formation of reactive oxygen intermediates and activated NF-kappaB and AP-1 in rat hepatocytes. HO-1 induction and hepatic damage were increased by BSO and only HO-1 induction was attenuated by the antioxidant N-acetylcysteine. HO-1 induction was also reduced by the cytochrome P-450 inhibitors ketoconazole and tranylcypromine, concomitantly with a significant decrease in the formation of diclofenac oxidative metabolites, which may give rise to reactive compounds. CONCLUSIONS: Acute treatment with diclofenac induces HO-1 in rodent hepatocytes. Induction is influenced by changes in the cellular redox states and by cytochrome P-450 activity and gives a new insight into the response of the liver to diclofenac. PMID- 12763372 TI - Gene expression profile in the regenerating rat liver after partial hepatectomy. AB - BACKGROUND/AIMS: When a loss of hepatic mass occurs, the expression of a large number of genes is either induced or altered, accompanying hepatocyte proliferation. In the present study, we made an in-house cDNA microarray containing 4608 elements (Liver chip), and analyzed extensively gene expression profiles of the regenerating liver after 70% partial hepatectomy (PHx) in rats. METHODS: RNAs were prepared from three rat livers at each time point (taken at 0, 6, 12, 18, 24, 48, 72 h, and 1 week after PHx). Using the liver chip, we performed large-scale analysis of gene expression during liver regeneration. Elements either up- or down-regulated more than twofold at one or more time points were selected. RESULTS: Among the 4608, 382 were identified. Using cluster analysis, we found great similarity between gene-expression profiles at 12 and 18 h after PHx as well as between 48 and 72 h after PHx. We also found that there are at least six distinct temporal patterns of gene expression in the regenerating rat liver after PHx. CONCLUSIONS: These results indicated that microarray analysis is a powerful approach for monitoring molecular events in the regenerating liver. PMID- 12763373 TI - Tumoral angiogenesis and tissue factor expression during hepatocellular carcinoma progression in a transgenic mouse model. AB - BACKGROUND/AIMS: The hypervascularity described in hepatocellular carcinoma varies according to the progression and the differentiation of the tumor, suggesting an angiogenic switch during tumor development. METHODS: We used a transgenic mouse model of hepatocellular carcinoma induced by the expression of SV40-T antigen, in which male mice developed hepatic tumors at various temporal and histological stages, whereas female mice remained tumor-free. We analyzed, by immunostaining and reverse transcription-polymerase chain reaction, factors involved in tumoral angiogenesis. RESULTS: We demonstrated that tumoral angiogenesis occurred before the development of diffuse hepatocarcinoma. We showed that some SV40-T-positive cells with an endothelial phenotype are involved in angiogenic processes, suggesting a partial vasculogenic mimicry. This tumoral angiogenesis is associated with platelet activation due to tissue factor expression in endothelial cells and invading macrophages. Normal and transgenic livers exhibited different pattern of expression of hypoxia-inducible factor 1 alpha (HIF-1alpha) and vascular endothelial growth factor (VEGF) mRNA. CONCLUSIONS: This model of hepatocellular carcinoma displays marked tumoral angiogenesis, with proliferation, remodeling and arterialization of hepatic sinusoids, probably associated with a partial vasculogenic mimicry. Abnormal angiogenesis observed in hepatocarcinoma was associated with platelet activation by tissue factor (TF) produced by endothelial cells and invading macrophages. In this transgenic model, HIF-1alpha, VEGF, and TF play a crucial role in tumoral angiogenesis. PMID- 12763374 TI - Constitutive over-expression of the insulin receptor substrate-1 causes functional up-regulation of Fas receptor. AB - BACKGROUND/AIMS: Insulin- and insulin growth factor-1 stimulated signaling through the insulin receptor substrate-1 (IRS-1) promotes hepatocellular proliferation and survival. IRS-1 over-expression in transgenic (Tg) mouse livers caused constitutive activation of Erk mitogen activated protein kinase (MAPK) and phosphatidylinositol-3 kinase (PI3K) resulting in significantly increased levels of DNA synthesis and larger hepatic masses relative to non-transgenic (non-Tg) littermates. However, the livers eventually ceased to grow but remained approximately 25% larger than non-Tg livers. We hypothesized that this growth homeostasis was achieved by parallel activation of pro-apoptosis pathways. METHODS: Since Fas-mediated apoptosis is a common mechanism of hepatocyte destruction, we investigated the potential role of Fas receptor as a regulator of hepatic mass in IRS-1 transgenic mice. RESULTS: Significantly increased Fas receptor levels were detected in the livers of IRS-1 Tg compared to non-Tg mice by Western blot analysis. Functional activation of Fas-receptor in IRS-1 Tg livers was demonstrated by increased hepatocellular apoptosis caused by intravenous injection of anti-Fas (Jo-2). CONCLUSIONS: These findings suggest that the increased growth caused by IRS-1 over-expression is balanced by constitutive activation of pro-death mechanisms. Failure of the IRS-1 Tg mice to develop liver cancer may be due to preservation of pro-growth, pro-death homeostasis mechanisms. PMID- 12763375 TI - A randomized study comparing lamivudine monotherapy after a short course of hepatitis B immune globulin (HBIg) and lamivudine with long-term lamivudine plus HBIg in the prevention of hepatitis B virus recurrence after liver transplantation. AB - BACKGROUND/AIMS: To compare the efficacy in preventing hepatitis B virus (HBV) recurrence of lamivudine vs. lamivudine plus hepatitis B immune globulin (HBIg) after a short course of HBIg and lamivudine in liver transplanted chronic hepatitis B patients. METHODS: Forty-six patients with HBV cirrhosis received lamivudine before liver transplantation and were then randomized to receive lamivudine plus HBIg for 1 month followed by lamivudine or both drugs for 17 months. RESULTS: Thirty-two patients were transplanted and 29 were randomized to receive combination therapy (15 cases) or lamivudine monotherapy (14 cases). HBV DNA was undetectable in all cases (17 induced by lamivudine therapy) at the time of liver transplantation. After 18 months of follow-up, all patients survived without HBV recurrence: hepatitis Bs antigen and HBV DNA were negative; however, HBV DNA was detected by polymerase chain reaction in four cases (three with HBIg plus lamivudine and one with lamivudine). Alanine aminotransferase levels were normal except in six cases (one HCV and two HDV coinfections). There were no drug related adverse events. CONCLUSIONS: Lamivudine monotherapy after a short course of lamivudine and HBIg is equally as efficacious in preventing HBV recurrence as HBIg plus lamivudine during the first 18 months after liver transplantation. This strategy is more economic and convenient to administer than long-term HBIg plus lamivudine. PMID- 12763376 TI - Lamivudine and 24 weeks of lamivudine/interferon combination therapy for hepatitis B e antigen-positive chronic hepatitis B in interferon nonresponders. AB - BACKGROUND/AIMS: Lamivudine is effective in treatment-naive patients with chronic hepatitis B, but its role in interferon nonresponders has not been described. We assessed lamivudine treatment, with or without added interferon, in patients with hepatitis B e antigen (HBeAg)-positive chronic hepatitis B who had failed interferon therapy previously. METHODS: Patients were randomized to lamivudine (100 mg) or placebo for 52 weeks or to a 24-week regimen of lamivudine plus interferon. Primary treatment comparisons were at week 52, with a 16-week posttreatment follow-up period. Measurements included histology (primary endpoint), HBeAg response, normalization of alanine aminotransferase, reduction of hepatitis B virus (HBV) DNA, and safety. RESULTS: Among 238 patients, histologic response was significantly more common in patients treated with lamivudine (52 versus placebo 25%, P=0.002) or the combination regimen (32%, P=0.01). HBeAg loss was also more common with lamivudine (33 versus 13 versus 21%), as were virologic and alanine aminotransferase responses. Among 28 subjects with HBeAg loss/seroconversion, 71% had durable responses 16 weeks posttreatment. CONCLUSIONS: Lamivudine for 52 weeks is as effective in interferon nonresponders as in previously reported treatment-naive patients; however, a combination of lamivudine for 24 weeks and interferon for 16 weeks was not effective in this population. PMID- 12763377 TI - Localization of swine hepatitis E virus in liver and extrahepatic tissues from naturally infected pigs by in situ hybridization. AB - BACKGROUND/AIMS: The objective of this study was to detect and localize the swine hepatitis E virus (HEV) in the liver and extrahepatic tissues from 20 pigs naturally infected with swine HEV. METHODS: cDNA probe 289 base pairs for swine HEV were generated by reverse transcription-polymerase chain reaction. In situ hybridization with a nonradioactive digoxigenin-labeled cDNA probe was used for the detection of swine HEV in formalin-fixed, paraffin-embedded tissues. RESULTS: When liver tissues from the pigs naturally infected with swine HEV were hybridized with the nonradioactive digoxigenin-labeled cDNA probe, a strong signal was seen in hepatocytes and bile duct. Positive hybridization signals were also detected in small and large intestine, lymph node, tonsil, spleen, and kidney. CONCLUSIONS: Swine HEV was detected primarily in the hepatocytes. Swine HEV may also replicate in tissues other than the liver. In situ hybridization described in the present study has greatly facilitated the application of in situ hybridization procedures to the clinical setting, thus allowing for the diagnosis of swine HEV infection while preserving the morphology of the tissue. PMID- 12763379 TI - Biliary cholesterol secretion: more lessons from plants? PMID- 12763378 TI - Mucosal humoral immune response to hepatitis C virus E1/E2 surface glycoproteins and HCV shedding in saliva and cervicovaginal fluids from chronically HCV infected patients. AB - BACKGROUND/AIMS: We herein focused on identifying biological factors possibly involved in non-parenteral transmission of hepatitis C virus (HCV), such as HCV excretion patterns and antibody-based immunity to the virus in saliva and/or cervicovaginal secretions (CVS). METHODS: Paired blood, saliva and cervicovaginal lavage samples were obtained from HCV-RNA plasma-positive hemoglobin (Hb) antigen and HIV-seronegative, HCV-seropositive males (n=13) and females (n=21). HCV specific antibodies were detected by ELISA in paired samples, and HCV-RNA was detected in cell-free and cell-associated body fluids. RESULTS: Antibodies to E1 HCV surface glycoprotein of the IgG and IgA isotypes showed similar, but less pronounced, profiles as IgG and IgA to E2. HCV-specific IgG and IgA in mucosal fluids likely originated predominantly from the systemic compartment, because HCV specific mucosal immunoglobulins involved primarily monomeric antibodies, including monomeric IgA, and because their specific activities for HCV antigens in corporeal fluids were similar to those in paired serum (Se). Viral shedding in saliva or CVS was restricted to cell-associated, non-replicating strand((+)) HCV RNA in 42% (12 out of 28) of saliva and in 19% (four out of 21) of cervicovaginal fluids. CONCLUSIONS: The association in body fluids of HCV-specific IgG, and to a lesser extent IgA, directed to E1/E2 surface glycoproteins (which may block critical steps of virus-cell interactions), of undetectable free viral RNA, and of occasional non-replicating cell-associated HCV, suggests a resulting poor infectivity of saliva or cervicovaginal fluid in chronically HCV-infected individuals. Taken together, these observations provide the basis for the low risk of non-parenteral transmission of HCV infection. PMID- 12763380 TI - Transcriptional profiling of liver regeneration: new approaches to an old trick! PMID- 12763382 TI - Treatment options for chronic hepatitis B not responding to interferon. PMID- 12763381 TI - Towards an improved and cost-saving prophylaxis of hepatitis B virus recurrence after liver transplantation? PMID- 12763383 TI - Osteoporosis in liver diseases and after liver transplantation. PMID- 12763384 TI - Activated protein C resistance acquired through liver transplantation and associated with recurrent venous thrombosis. AB - We report a new case of recurrent, extra-hepatic, deep vein thrombosis occurring after orthotopic liver transplantation for hepatocellular carcinoma complicating 'mixed' alcoholic and post-hepatitic C cirrhosis. Coagulation tests showed activated protein C resistance. The patient's genomic DNA was negative for the factor V Leiden mutation. Analysis of the grafted liver DNA showed that the donor was a heterozygous carrier of the factor V Leiden mutation and that the recipient's activated protein C resistance was acquired through the transplantation. Screening of candidate liver donors for a prothrombotic tendency is controversial. However, this case suggests that patients who develop venous thrombosis after liver transplantation should be screened for thrombophilic abnormalities, bearing in mind that genetic abnormalities which do not affect clotting test results, such as the G20210A mutation in the factor II gene, can only be diagnosed by testing the donor or graft. PMID- 12763385 TI - Hepatic cyst rupture after a coughing fit. PMID- 12763386 TI - Severe migraine headaches are caused by ribavirin but not by interferon alpha-2B in combination therapy for chronic hepatitis C. PMID- 12763387 TI - Die another day. PMID- 12763389 TI - Direct and retrospective assessment of factors contributing to compulsive buying. AB - Compulsive buying is a disorder that has begun to receive attention from researchers in recent years. The results of a handful of studies suggest that compulsive buying occurs in response to negative emotions and results in a decrease in the intensity of the negative emotions. In this investigation, we used interview and self-monitoring methods to evaluate the antecedents and consequences of compulsive buying in a sample of women who met criteria for compulsive buying on the compulsive buying scale (J. Consumer Res. 19 (1992) 459). As a group, the participants reported negative emotions as the most common antecedents to compulsive buying, and euphoria or relief from the negative emotions as the most common consequence of compulsive buying. These findings were consistent across the interview and self-monitoring assessment methods. The implications for assessment and treatment are discussed. PMID- 12763390 TI - Catastrophic worry in primary insomnia. AB - The present study aimed to provide an empirical test of the proposal that catastrophic worry about the consequences of not sleeping is common among patients with primary insomnia and serves to maintain the sleep disturbance. It was predicted that relative to good sleepers, patients with primary insomnia would catastrophize more and that catastrophizing would be associated with increased negative affect and increased perception of threat. A 'catastrophizing interview' was administered to 30 patients with primary insomnia and 30 good sleepers. Consistent with the predictions, the insomnia patients generated more catastrophes about the consequences of not sleeping and gave higher likelihood ratings than good sleepers. For the insomnia group, but not the good sleepers, the catastrophizing interview was associated with increased anxiety and discomfort. The limitations of the study, possibilities for future research, and clinical implications of these findings are discussed. PMID- 12763391 TI - Screening for generalized anxiety disorder using the Penn State Worry Questionnaire: a receiver operating characteristic analysis. AB - The present study examined the usefulness of the Penn State Worry Questionnaire (PSWQ) as a means of screening for generalized anxiety disorder (GAD). Using receiver operating characteristic analyses, the accuracy of the PSWQ in screening for GAD was examined in both clinical and analogue diagnosed GAD samples. Given high comorbidity between GAD and other emotional disorders, we also investigated the usefulness of the PSWQ in selecting non-cases of GAD that were also free of PTSD, social phobia, or depression versus non-cases of GAD that met criteria for one of these conditions. The overall usefulness of the PSWQ as a screening device is discussed. PMID- 12763392 TI - Development and characteristics of a well-being enhancing psychotherapeutic strategy: well-being therapy. AB - This article describes the main characteristics and technical features of a novel psychotherapeutic strategy, well-being therapy. This paper outlines the background of its development, the structure of well-being therapy, its key concepts and technical aspects. Well-being therapy is based on Ryff's multidimensional model of psychological well-being, encompassing six dimensions: autonomy, personal growth, environmental mastery, purpose in life, positive relations and self-acceptance. The goal of this therapy is improving the patients' levels of psychological well-being according to these dimensions, using cognitive-behavioral techniques. It may be applied as a relapse-preventive strategy in the residual phase of affective (mood and anxiety) disorders, as an additional ingredient of cognitive-behavioral packages, in patients with affective disorders who failed to respond to standard pharmacological or psychotherapeutic treatments and in body image disturbances. The clinical studies supporting its efficacy are illustrated. PMID- 12763393 TI - The Dental Cognitions Questionnaire in CBT for dental phobia in an adolescent with multiple phobias. AB - A case of an adolescent boy with multiple phobias who was treated successfully for his dental phobia is described to illustrate the clinical utility of the Dental Cognitions Questionnaire (DCQ) in aiding effective cognitive-behavior therapy. The client showed drops in dental anxiety that coincided with the use of the DCQ in cognitive restructuring, and there was a close correlation between dental cognitions and degree of dental anxiety over the time-course of therapy and follow up. PMID- 12763394 TI - Cognitive change processes in a group cognitive behavior therapy of depression. AB - The present study attempted to examine the causal relationships among changes in automatic thoughts, dysfunctional attitudes, and depressive symptoms in a 12-week group cognitive behavior therapy (GCBT) program for depression. In all, 35 depressed patients attending the GCBT program were monitored with the Automatic Thoughts Questionnaire, Dysfunctional Attitudes Scale, and Beck Depression Inventory at the pre-treatment, 4th and 8th sessions, and post-treatment. The results were as follows: (1). GCBT reduces negative cognitions; (2). changes in automatic thoughts and dysfunctional attitudes lead to change in depressive symptoms; and (3). automatic thoughts play a mediating role between dysfunctional attitudes and depression. The findings taken as a whole support the Causal Cognition Model of depression. PMID- 12763395 TI - 2002 Consensus Conference on Management of Obesity. PMID- 12763396 TI - Epidemiology and consequences of obesity. PMID- 12763397 TI - Body weight regulation and obesity. PMID- 12763399 TI - Medical management of obesity: present and future therapy. PMID- 12763398 TI - Lifestyle modification in the management of obesity. PMID- 12763400 TI - Open and laparoscopic surgical modalities for the management of obesity. PMID- 12763401 TI - Treatment of obesity. PMID- 12763403 TI - Overview--current clinical and preclinical use of robotics for surgery. PMID- 12763404 TI - Robots in surgery: advanced gastrointestinal applications and credentialing. PMID- 12763405 TI - Outcome after pancreaticoduodenectomy for periampullary cancer: an analysis from the Veterans Affairs National Surgical Quality Improvement Program. AB - The aim of this study is to define the risk factors that predict adverse outcomes for patients undergoing pancreaticoduodenectomy for periampullary cancer in the Department of Veterans Affairs Healthcare System (VA). The VA National Surgical Quality Improvement Program prospectively collected data on 462 patients undergoing pancreaticoduodenectomy in 123 VA medical centers from 1990 to 2000. Independent variables included 68 preoperative and 12 intraoperative variables. The main outcome measures were 30-day postoperative mortality and morbidity, as measured by a set of 20 pre-defined complications. Predictive models for 30-day morbidity and mortality were constructed using logistic regression analysis. The 30-day morbidity rate was 45.9% (212/462). The 30-day postoperative mortality rate was 9.3% (43/462). Significant predictors of mortality included: preoperative serum albumin, American Society of Anesthesiologists classification, preoperative bilirubin >20mg/dl, and operative time. The use of preoperative biliary tract instrumentation did not predict postoperative death or septic complications. This study provides a set of preoperative risk factors that are predictive of adverse outcome following pancreaticoduodenectomy. These factors may assist in patient selection for this procedure and are likely to facilitate risk-adjusted comparison of pancreaticoduodenectomy outcomes between different health care systems. PMID- 12763406 TI - Transcystic common bile duct exploration in the management of patients with choledocholithiasis. AB - Common bile duct stones are found in approximately 16% of patients undergoing laparoscopic cholecystectomy. If the diagnosis of choledocholithiasis is made at the preoperative workup, it is common practice to refer the patient for endoscopic retrograde cholangiography and endoscopic sphincterotomy. However, if the diagnosis is established during intraoperative cholangiography, the surgeon is confronted with a therapeutic dilemma-that is, the choice between laparoscopic common bile duct exploration, conversion to open surgery, or postoperative endoscopic sphincterotomy. We have opted to treat patients with choledocholithiasis in only one session during the laparoscopic cholecystectomy; we use the transcystic common bile duct exploration technique employing the choledochoscope. We report our early experience in terms of success of stone removal, operative time, morbidity and mortality, and length of hospital stay. From 1992 to 2002, we performed 350 laparoscopic cholecystectomies. Selective cholangiography was used in 105 patients (30%); 40 of them were found to have common bile duct stones, for an incidence of 11.4%. Among this group, we performed laparoscopic transcystic common bile duct exploration in all but six patients. Our success rate for stone removal was 94.1% (32 of 34 patients), with only two failures related to multiple stones and impaction at the ampulla, for a conversion rate of 5.8%. The mean operative time was 120+/-40 minutes. The morbidity rate was 8.8%, and there were no deaths. Length of hospital stay was 24 to 48 hours. Mean recovery time was 7 days, and time to return to work was 15+/-3 days. We concluded that most of the patients with common bile duct stones found during laparoscopic cholecystectomy could be treated successfully by means of the transcystic technique with choledochoscopy, with no increase in morbidity or mortality and a shortened hospital stay and recovery time, similar to patients who undergo only laparoscopic cholecystectomy. On the basis of our results, we recommend that this method become the primary strategy in the great majority of patients with common bile duct stones found during intraoperative cholangiography. PMID- 12763407 TI - Biliary obstruction reduces hepatic killing and phagocytic clearance of circulating microorganisms in rats. AB - Septic complications are common in patients with biliary obstruction. This is thought to be related, in part, to dysfunction of the hepatic reticuloendothelial system (RES). It has been reported that nearly 80% of circulating microorganisms are phagocytosed and killed within the liver and that clearance of circulating pathogens is significantly impaired in patients with jaundice. However, the effect of biliary obstruction specifically on phagocytic killing within the liver is less well described. Therefore this study was designed to quantify the effect of biliary obstruction, simultaneously and discriminately, on two important components of hepatic RES function (phagocytosis and phagocytic killing). Rats were divided into three experimental groups: control, sham, and jaundiced (common bile duct ligation). At 7, 10, 14, and 21 days after operation, E. coli labeled with both 125I and 51Cr were injected intravenously. Using the previously validated double-labeled in vivo E. coli technique, hepatic phagocytic clearance (HPC), hepatic killing efficiency (HKE), and net hepatic killing (NHK) were measured. Common bile duct ligation resulted in a significant decrease in the HPC of E. coli 10, 14, and 21 days postoperatively. Similarly, HKE was significantly decreased in jaundiced animals by postoperative day 10, but returned to baseline values by day 14. The net effect of these changes in HPC and HKE values were reflected in a significant reduction in NHK in jaundiced animals. Results of the present study suggest that obstructive jaundice impairs both phagocytosis and phagocytic killing within the liver. These findings may help to explain the susceptibility of patients with biliary tract obstruction to the morbidity and mortality of septic complications. PMID- 12763408 TI - An intravital model to monitor steps of metastatic tumor cell adhesion within the hepatic microcirculation. AB - Organ-specific tumor cell adhesion within the microcirculation of host organs is an important step in the metastatic cascade. Circulating tumor cells have to adhere within the microcirculatory vessels, quickly stabilize their adhesion and probably leave the circulation to avoid toxic effects of hydrodynamic shear forces of circulating blood. Using intravital fluorescence microscopy we established a new model for the intravital observation of colon carcinoma cell adhesion within the hepatic microcirculation. HT-29 (human) and CC531 (rat) colon carcinoma cells were fluorescence labeled using CalceinAM. Single cell suspensions were injected intraarterially in Sprague-Dawley rats. Using intravital fluorescence microscopy adhesive interactions of circulating tumor cells within the hepatic microcirculation were observed at the liver surface. These interactions were analyzed regarding their time course and the localization within the vascular tree. Autofluorescence of liver parenchyma was sufficient for distinction of hepatic sinusoids. Intravital microscopy enabled the differentiation of early events in adhesion formation within hepatic sinosoids, adhesion stabilization, and extravasation of the tumor cells into the liver parenchyma. Tumor cell adhesion occurred almost exclusively within sinusoidal capillaries; however, the diameter of these vessels was usually larger than that of the tumor cells leaving remaining perfused lumen of the capillaries. Colon carcinoma cells rapidly migrated into the liver parenchyma after successful adhesion within the sinusoids. In contrast to common endpoint assays of the metastatic cascade, this in vivo model allows investigations of metastatic colon carcinoma cell adhesion within the liver microcirculation as specific steps during the formation of hematogenous metastasis and their underlying mechanisms. PMID- 12763409 TI - Identification of optimal harvest sites of ileal stem cells for treatment of bile acid malabsorption in a dog model. AB - Ileal mucosal stem cells expressing the sodium-dependent ileal bile acid transporter (IBAT) have been successfully transplanted into the jejunum of rodents in projects aimed at creating a "neoileum" to treat bile acid malabsorption. To find optimal harvest sites for a dog model of stem cell transplantation, the exact location of peak IBAT expression in the donor ileum needs to be known. We therefore mapped IBAT function, IBAT mRNA, and IBAT protein in the ileum of Beagle dogs (N=3). Mucosal samples were taken every 5 cm in the ileum and every 20 cm in the jejunum of each dog. Sodium-dependent (active) and sodium-independent (passive) taurocholate uptake rates were measured using a standardized everted sleeve technique. IBAT mRNA concentrations were determined by semiquantitative reverse transcriptase-polymerase chain reaction and IBAT protein concentrations by fluorometric immunohistochemical analysis. The small bowel measured 208+/-17 cm (mean+/-standard error of the mean). Active and passive uptake rates were found to follow distinct distribution curves. Significant active uptake was seen only at the terminal 50 cm and peaked at 479+/ 176 pM/mm(2). Depending on location, active uptake accounted for approximately half of the total uptake. IBAT mRNA and protein distributions corroborated uptake curves. The terminal 10 to 50 cm of ileum has the highest bile acid uptake capacity. This short segment appears to be the most promising donor site for ileal stem cell transplants to create a "neoileum" in dogs. PMID- 12763410 TI - Institutional variations in the management of patients with acute appendicitis. AB - The purpose of this study was to evaluate institutional differences in preoperative workup, operative approach, complications, and cost in patients with acute appendicitis. A retrospective chart review was performed of all adults operated on for acute appendicitis from June 1999 to November 2000 at the University of New Mexico Hospital (UNMH) and Stanford University Medical Center (SUMC). Variables compared included age, race, sex, duration of symptoms, type of symptoms, results of radiographic evaluation, time from emergency room to operating room, operative approach (open vs. laparoscopic), operative time, length of hospital stay, pathologic findings, and complications. Statistical analysis was performed by means of Fisher's exact test. A total of 154 appendectomies were performed for acute appendicitis at UNMH and 165 at SUMC. Statistically significant differences were found at UNMH vs. SUMC in time from emergency room to operating room (9.1 hours vs. 13.7 hours; P<0.001), operative approach (48% laparoscopic vs. 29% open; P<0.001), and negative appendectomy rate (13% vs. 4.8%; P<0.001). There were no differences in the perforation rate or other complications. Cost analysis showed that $56,744 more was spent at UNMH for the additional negative appendectomy operations, whereas $99,842 more was spent at SUMC for the additional CT scans. Institutional differences in the management of patients with acute appendicitis can result in significant differences in cost without clinically significant differences in outcome. The use of clinical examination and laparoscopy as diagnostic modalities instead of CT scanning resulted in a more cost-effective approach. PMID- 12763411 TI - Reduction in serum pepsinogen I after Roux-en-Y gastric bypass. AB - The excluded stomach after Roux-en-Y gastric bypass (RYGBP) cannot be readily examined by endoscopy for obvious anatomic reasons. Thus it is difficult to monitor possible changes in the gastric mucosa. However, the type and severity of gastritis can now be assessed by a combination of serologic tests: pepsinogen I and antibodies to Helicobacter pylori and H,K-ATPase. Morbidly obese patients were examined before and 1 to 4 years after surgery. A group of 34 patients (mean age 39 years, BMI 44 kg/m(2)) underwent RYGBP; another group of 30 patients (mean age 42 years, BMI 44 kg/m(2)) had simple gastric restriction and served as control subjects. All patients, except one in the control group, had normal titers of pepsinogen I before surgery. One year after RYGBP, pepsinogen I levels were significantly reduced, as compared to the control group (P<0.0001), and remained low throughout the study. The control group had stable pepsinogen I levels. In both groups, few patients had increased titers of H. pylori or H,K ATPase antibodies, but these abnormalities remained unchanged. Low pepsinogen I levels, similar to those we observed in our RYGBP patients, have been linked to chronic atrophic gastritis. However, the absence of food stimulation in the excluded stomach could also be a reason for the low pepsinogen I levels. PMID- 12763413 TI - Surgical management of pancreaticocutaneous fistula. AB - Although enteric drainage of the fistula tract is a widely accepted treatment for pancreaticocutaneous fistula, few data have been published on the outcome of this procedure. We conducted a retrospective chart review of 30 patients with pancreaticocutaneous fistula who underwent surgical management at a single institution over a 13-year period. The operative morbidity rate was 30%. Overall the incidence of recurrent ductal leaks requiring further intervention was 23%. Six of seven patients who had a recurrence had an ongoing inflammatory pathology, and three of seven had pancreas divisum. Recurrence was most likely when cystenterostomy was used. Enteric drainage of pancreaticocutaneous fistulas is not always curative. Fistulojejunostomy gives a better outcome than cystenterostomy. Recurrence may be expected in patients with continuing inflammatory ductal pathology. PMID- 12763412 TI - Benign nonampullary duodenal neoplasms. AB - Benign duodenal neoplasms (BDNs) are uncommon, and their optimal management remains undefined. We analyzed all cases of BDN treated at our institution during a 10-year period (January 1990 through January 2000). Data are expressed as median (range). Sixty-two patients were treated for BDNs. The results of histologic examination of their lesions were as follows: 36 adenomas, eight Brunner's gland tumors, 10 inflammatory polyps, two hamartomas, and six others. Forty-seven patients were treated nonoperatively, and 15 patients underwent surgery. Lesion characteristics leading to surgical intervention included large polyp diameter and submucosal penetration detected on endoscopic ultrasound imaging. There were no treatment-related deaths. Major morbidity occurred in 2% of patients who underwent endoscopic resection and in 33% of patients who underwent surgery (P = 0.002). Among patients treated for adenomas, seven (19.4%) had a recurrence at a median of 12 (4 to 48) months. Most BDNs can be managed with minimal morbidity using endoscopic techniques. Systematic follow-up of patients treated for adenomas is required. PMID- 12763414 TI - Histologic findings of gallbladder mucosa in 87 patients with morbid obesity without gallstones compared to 87 control subjects. AB - Obesity is an important risk factor for the development of gallstones. The purpose of this study was to determine histologic alterations in the gallbladder mucosa and the prevalence of gallstone disease in patients with severe and morbid obesity compared to histologic findings in the gallbladder mucosa of control subjects. Two groups were studied: 125 severely obese patients (38 with and 87 without gallstones) and 87 control subjects. Ultrasonography was performed in all of them before surgery. During surgery, cholecystectomy was performed in 87 obese patients with a "normal" gallbladder and in all 87 control subjects. Specimens were immediately sent for histologic analysis. The prevalence of gallstones was twice as high among obese women compared to obese men (P < 0.001). Normal gallbladder mucosa was found in 28.7% of obese women compared to 34.2% of control women (P > 0.59). Findings were similar among the men. The most frequent histologic abnormality in the gallbladder mucosa among obese women was cholesterolosis (37%), followed by chronic cholecystitis and cholesterolosis (18%), with frequencies of 23% and 12%, respectively, in control women (P > 0.1). Among men, a similar proportion of histologic abnormalities was seen in obese men and control subjects. In our population of obese patients compared to control subjects, a similarly high proportion of histologic abnormalities of the gallbladder mucosa was found in the absence of stones. These findings could have been attributed to the fact that the Chilean population has a high incidence of gallstones. PMID- 12763415 TI - Laparoscopic vs. open biliopancreatic diversion with duodenal switch: a comparative study. AB - Biliopancreatic diversion with duodenal switch (BPD-DS) is a well-known emerging open procedure that appears to be as effective as other bariatric operations and has been shown to provide excellent long-term weight loss. Therefore we looked at the safety and efficacy of the laparoscopic BPD-DS procedure compared to open BPD DS in superobese patients (body mass index >60). A retrospective study of 54 superobese patients (body mass index >60) was carried out from July 1999 to June 2001: laparoscopic BPD-DS in 26 patients and open BPD-DS in 28 patients. Median preoperative body weight was 189.8 kg (range 155.1 to 271.2 kg) in the laparoscopic BPD-DS group and 196.5 kg (range 160.3 to 298.9 kg) in the open BPD DS group. Median body mass index was 66.9 kg/m(2) in the laparoscopic group and 68.9 kg/m(2) in the open group. The two groups were compared by means of the unpaired t test, which yielded the following results: Major morbidity occurred in six patients (23%) in the laparoscopic BPD-DS group and in five patients (17%) in the open BPD-DS group (P = 0.63). There were two deaths in the laparoscopic BPD DS group (7.6% mortality) and one death (3.5% mortality) in the open BPD-DS group (P = 0.51). Preoperative comorbidity was improved in eight patients in the laparoscopic BPD-DS group and two patients in the open BPD-DS group (P < 0.02). Laparoscopic BPD-DS is a technically feasible procedure that results in effective weight loss similar to the open procedure. However, both open and laparoscopic BPD-DS procedures are associated with appreciable morbidity and mortality in the superobese population. Additional studies are needed to determine the best surgical treatment for superobesity. PMID- 12763416 TI - Laparoscopic colectomy in obese and nonobese patients. AB - Obese patients carry a higher risk of wound complications and cardiopulmonary complications along with a higher incidence of comorbidity, all of which have the potential to affect outcome after a variety of surgical procedures. The data regarding outcomes after laparoscopic colectomy in obese and nonobese patients are limited. The purpose of this report was to compare the outcome of laparoscopic bowel resection in obese and nonobese patients. All patients prospectively entered into a laparoscopic bowel resection database from March 1999 to December 2001, who underwent a segmental colectomy for any pathologic condition, were analyzed. Patients with a body mass index above 30 were defined as obese, and patients with a body mass index below 30 were defined as nonobese. Data collected included age, sex, duration of operation, body mass index, American Society of Anesthesiologists score, operative procedure, diagnosis, complications relating to length of hospital stay, mortality, and readmission within 30 days of discharge. Statistical analysis consisted of Student's t test and chi-square analysis where appropriate, with significance set at P < 0.05. A total of 260 patients were evaluated (201 [77.3%] in the nonobese group and 59 [22.7%] in the obese group). There were no significant differences between the two groups with respect to age, sex, operative procedure, length of hospital stay, or readmission rates. The obese group had significantly more conversions to an open procedure (23.7% vs. 10.9%), a longer operative duration (109 minutes vs. 94 minutes), a higher morbidity rate (22% vs. 13%) and a higher anastomotic leakage rate (5.1% vs. 1.2%). This large experience with laparoscopic colectomy for a variety of conditions demonstrates that despite higher conversion rates, an increased risk of pulmonary complications, and anastomotic leakage rates in obese laparoscopic patients that parallel those of open surgery, laparoscopic colectomy can be performed safely in both obese and nonobese patients with the similar benefit of a shorter hospital stay in both groups. PMID- 12763417 TI - Home total parenteral nutrition: an alternative to early surgery for complicated inflammatory bowel disease. AB - This paper examines the safety and feasibility of providing short-term, in-home total parenteral nutrition (TPN) for patients with inflammatory bowel disease (IBD) for whom the alternative is prolonged hospitalization or early surgery. The records of all patients with IBD who were receiving temporary home TPN between June 1996 and July 2000 were reviewed. A quality-of-life phone interview was conducted at the time of review. Fifteen patients (11 men and 4 women) were identified whose average age was 35 years. The underlying diagnosis was Crohn's disease in 10 and ulcerative colitis in five. The indications for home TPN were complex internal fistulas and resolving sepsis in two, postoperative septic complications (anastomotic leak/enterocutaneous fistula) in five, high-output proximal stomas in four, prolonged ileus/partial obstruction in three, and spontaneous enterocutaneous fistula in one. The average duration of home TPN was 75 days (range 7 to 240 days). Two patients (13%) failed home TPN (1 with uncontrolled sepsis; 1 with dehydration) and were readmitted to the hospital. Home TPN was discontinued in one patient whose enterocutaneous fistula failed to heal with nonoperative treatment. Home TPN was successful in 12 patients (80%): eight (53%) who underwent planned definitive surgery and four (27%) whose conditions resolved without surgery. Complications of home TPN were line sepsis and pulmonary aspergillosis in one patient. All patients preferred home TPN to further hospitalization and reported good or excellent quality of life at home. Home TPN is a safe alternative to prolonged hospitalization or early surgery in patients with complicated IBD. PMID- 12763418 TI - Outcome of patients undergoing ileal pouch-anal anastomosis for left-sided chronic ulcerative colitis. AB - Chronic ulcerative colitis is not a uniform disease entity because the clinical pattern and disease characteristics differ on the basis of the anatomic location of the inflammation. The aim of this study was to compare the preoperative characteristics, postoperative complications, and long-term functional outcome of ileal pouch-anal anastomosis (IPAA) in patients with left-sided colitis to those same characteristics in patients with pancolitis. Between 1990 and 1996, a total of 565 patients underwent IPAA for chronic ulcerative colitis at our institution. Of these, 111 patients were determined to have left-sided involvement, whereas 283 patients had pancolitis. The mean age at surgery was greater in the patients with left-sided colitis (37 years vs. 34 years, P = 0.01), and the mean duration of disease (8.7 years vs. 7.7 years, P = 0.05) tended toward a significant difference between the left-sided colitis and pancolitis groups. The complication rates were similar with the exception of small bowel obstructions, for which there was a higher incidence in the group with left-sided colitis (27% vs. 13%, P = 0.002) at 5 years. The incidence of pouchitis (43% vs. 39%) at 5 years was comparable. Long-term functional results and quality-of-life assessment did not show any significant differences between the two groups. We were unable to detect any correlation between the extent of colon involvement and the subsequent incidence of pouchitis, long-term pouch function, and quality of life. Patients with left-sided colitis were older, had a relatively longer duration of disease, and were at increased risk for postoperative small bowel obstruction as compared to patients with pancolitis. PMID- 12763420 TI - Waking up to sleeping sickness. AB - Devastating epidemics of human African trypanosomiasis are currently re-emerging in many sub-Saharan countries. In the past three decades, clinical research into this important disease has been neglected, as have urgently needed initiatives on drug development, disease surveillance and vector control. Recent impetus has aimed to provide a free supply of antitrypanosomal drugs, to develop a new orally active trypanocidal agent and to attack the tsetse vector with modern technology. In addition, pan-African initiatives to co-ordinate control efforts have begun. These all provide some hope for the future, but they might not be enough to reverse the resurgence of this deadly disease in the heart of Africa. PMID- 12763419 TI - Neuronal adrenergic and muscular cholinergic contractile hypersensitivity in canine jejunum after extrinsic denervation. AB - Extrinsic denervation may be responsible for motor dysfunction after small bowel transplantation. The aim of this study was to examine the role of extrinsic innervation of canine jejunum on contractile activity. An in vitro dose response of cholinergic and adrenergic agonists was evaluated in canine jejunal strips of circular muscle at 0, 2, and 8 weeks in a control group and after jejunoileal extrinsic denervation (EX DEN). Neurons in circular muscle were quantitated by means of immunohistochemical techniques. Adrenergic and cholinergic responses did not differ at any time in the control group. However, at 2 and 8 weeks, extrinsic denervation caused an increased sensitivity to the procontractile effects of the cholinergic agonist bethanechol at the level of the smooth muscle cells, and increased sensitivity to the inhibitory effects of the adrenergic agent norepinephrine mediated at the level of the enteric nervous system. Immunohistochemical analysis showed a reduction in all neurons and a complete lack of adrenergic fibers in the EX DEN group after 2 and 8 weeks. Extrinsic denervation induces enteric neuronal cholinergic and adrenergic smooth muscle hypersensitivity in canine jejunal circular muscle. PMID- 12763421 TI - The hidden faces of hemozoin and its dangerous midwives. PMID- 12763422 TI - The mysteries of hemoglobin degradation in Plasmodium falciparum-infected erythrocytes. PMID- 12763424 TI - Strategic options for global lymphatic filariasis elimination. PMID- 12763425 TI - Switching rates of Plasmodium falciparum var genes: faster than we thought? AB - Plasmodium falciparum undergoes antigenic variation by switching the expressed erythrocyte membrane protein (PfEMP)1. This family of proteins plays an important role in the development of chronic, recrudescent P. falciparum malaria, acquired immunity and severe malaria. However, little is known about the switching mechanism or switching rates in the human host. Here, we estimate the switch rate of var genes, using recently published data describing the var gene transcripts detected in blood taken from human volunteers during acute P. falciparum infections and a mathematical model of the in-host dynamics. The overall switch rate of PfEMP1 predicted during the initial stage of infection ( approximately 18% switching parasites per generation) is much higher than previously reported. The implications of the predicted switching rates are discussed. PMID- 12763426 TI - The ins, outs and roundabouts of malaria. AB - The malaria parasite Plasmodium falciparum is a complex eukaryote parasite with a dynamic pattern of genomic expression, enabling it to exploit a series of different habitats in human and mosquito hosts. In the human bloodstream, the parasite grows and multiplies within red blood cells and modifies them in various ways to gain nutrients and combat the host's defences, before escaping and invading new red blood cells by a multi-step process. These events are reflected in the constantly changing structure of the organism during the red blood cell cycle. PMID- 12763427 TI - Speculations on the origins of Plasmodium vivax malaria. AB - It is likely that Plasmodium vivax diverged approximately 2 million years ago from a group of malaria parasites which are now endemic in monkeys and apes in southern Asia. In those times, primates were spread throughout most of Eurasia and Africa, indicating an Old World location, but nothing more precise, for the place of divergence of P. vivax. From approximately 1 million years ago, the Ice Ages would have isolated human malaria, including P. vivax, into humid temperate or warm climate refuges around the Mediterranean, in sub-Saharan Africa and in south and east Asia. As there appears to be no record of humans in south and east Asia from 100,000 to 60,000 years ago, they might not have passed on their parasites, including P. vivax, to modern humans entering the region after this time. Today, all P. vivax might be descended from parasites which infected human populations in the Mediterranean region and in sub-Saharan Africa during the last Ice Age, between 100,000 and 20,000 years ago. Evidence for the latter is provided by the presence of very high frequency RBC Duffy negativity in sub Saharan Africa. PMID- 12763428 TI - The genetic diversity of Plasmodium vivax populations. AB - Little is known of the genetic diversity and population structure of Plasmodium vivax, a debilitating and highly prevalent malaria parasite of humans. This article reviews the known polymorphic genetic markers, summarizes current data on the population structure of this parasite and discusses future prospects for using knowledge of the genetic diversity to improve control measures. PMID- 12763429 TI - The Plasmodium vivax genome sequencing project. AB - With the successful completion of the project to sequence the Plasmodium falciparum genome, researchers are now turning their attention to other malaria parasite species. Here, an update on the Plasmodium vivax genome sequencing project is presented, as part of the Trends in Parasitology series of reviews expanding on various aspects of P. vivax research. PMID- 12763430 TI - Pentamidine uptake and resistance in pathogenic protozoa: past, present and future. AB - Diamidines, and pentamidine in particular, have a long history as valuable chemotherapeutic agents against infectious disease. Their selectivity is due mostly to selective accumulation by the pathogen, rather than the host cell; and acquired resistance is frequently the result of changes in transmembrane transport of the drug. Here, recent progress in elucidating the mechanisms of diamidine transport in three important protozoan pathogens, Trypanosoma brucei, Leishmania and Plasmodium falciparum, is reviewed, and the implications for drug resistance are discussed. PMID- 12763431 TI - Anatomical and biomechanical mechanisms of subacromial impingement syndrome. AB - Subacromial impingement syndrome is the most common disorder of the shoulder, resulting in functional loss and disability in the patients that it affects. This musculoskeletal disorder affects the structures of the subacromial space, which are the tendons of the rotator cuff and the subacromial bursa. Subacromial impingement syndrome appears to result from a variety of factors. Evidence exists to support the presence of the anatomical factors of inflammation of the tendons and bursa, degeneration of the tendons, weak or dysfunctional rotator cuff musculature, weak or dysfunctional scapular musculature, posterior glenohumeral capsule tightness, postural dysfunctions of the spinal column and scapula and bony or soft tissue abnormalities of the borders of the subacromial outlet. These entities may lead to or cause dysfunctional glenohumeral and scapulothoracic movement patterns. These various mechanisms, singularly or in combination may cause subacromial impingement syndrome. PMID- 12763432 TI - Reaction time and movement time in patients with carpal tunnel syndrome: an electromyographic study. AB - OBJECTIVE: To measure the motor performance of the flexor pollicis brevis, first dorsal interosseus, and flexor pollicis longus muscles using the parameters electromyographic muscle reaction time and movement time in patients with carpal tunnel syndrome during key turning function. METHOD: A total of 37 female patients with right hand pain were divided into two groups: 21 with an electrophysiologically confirmed diagnosis of carpal tunnel syndrome (Group I), and 16 with normal electrophysiological findings (Group II). The reaction time and movement time values obtained by surface electromyography of the right flexor pollicis brevis, first dorsal interosseus, and flexor pollicis longus muscles were compared between groups. RESULTS: Of the 21 patients in Group I, 15 had mild, four moderate, and two severe median nerve lesions. No significant difference between groups was found in the electromyographic reaction time and movement time values of the right flexor pollicis brevis, first dorsal interosseus, and flexor pollicis longus muscles during key turning (P>0.05). There was also no significant difference in electromyographic reaction time and movement time between six patients with severe or moderate carpal tunnel syndrome and 15 patients with mild carpal tunnel syndrome in Group I (P>0.05). Sixteen patients in Group II did not have carpal tunnel syndrome. CONCLUSION: The results indicate that patients with and without carpal tunnel syndrome have similar values of reaction time and movement time of the flexor pollicis brevis, first dorsal interosseus, and flexor pollicis longus muscles during the pinch grasp function of the hand. RELEVANCE: Muscle reaction time and movement time, and hence the motor performance in electrophysiologically confirmed carpal tunnel syndrome patients appear to have similar parameters in patients with hand pain whose diagnosis were not confirmed with electrophysiologically. PMID- 12763433 TI - Effect of simulated rock climbing finger postures on force sharing among the fingers. AB - OBJECTIVE: To study the forces applied by each finger in different joint postures simulating rock climbing gripping postures. DESIGN: Subjects in sitting posture applied fingertip forces perpendicular to horizontal force sensors in three different finger postures. BACKGROUND: Data provided by the literature indicate that middle and ring finger are commonly injured. However, no quantitative assessment of the forces applied by each finger related to the joint postures has been made. METHODS: Six elite rock climbers performed finger flexion in a single finger task and a four-finger task. The tests were conducted in an extended posture, a curved posture (the joints belonging to the finger were flexed) and an intermediate posture (the joints were flexed, except the distal one which was fully extended). Each fingertip force was expressed in absolute value and in percentage of the maximal force capacity of the finger. RESULTS: The greater force was applied by the middle finger (20.8 N), whatever the posture. The relative involvement amounted to 105% for the ring finger in the curved posture. CONCLUSIONS: The great force applied by the middle finger and the great relative involvement of the ring finger in the curved posture seem to be the main factors of injuries of these fingers. RELEVANCE: The analysis of force sharing among the fingers during different joint postures mimicking rock climbing is essential to a better understanding of finger injuries. PMID- 12763434 TI - No effects of cervical spine motion on cranial dura mater strain. AB - OBJECTIVE: The aim of this study was to analyze the effects of cervical spine motion on cranial dura mater length variations in anatomical specimens using high resolution linear displacement transducers. We hypothesized that transducer resolution was sufficient to measure dura mater length changes if they occurred during cervical spine motion. DESIGN: Cranial dura mater strain was measured using differential variable reluctance transducers during cervical spine motion in 11 formaldehyde-fixed whole-body anatomical specimens (mean age: 82 years). BACKGROUND: Several theories hypothesize that functional maneuvers carried out on the spine have an effect on intra-cranial structures due to the supposed continuity of spinal and cranial dura mater. The displacements of the spinal dura mater are supposed to be transmitted to the cranial dura mater. METHODS: Eleven anatomical specimens were used. Each specimen (positioned supine) was provided with three openings in the skull (frontal and parietal regions), leaving the dura mater intact. A differential variable reluctance transducer was inserted in frontal or sagittal orientation in the dura mater exposed in each opening. Strain was recorded during cyclic motions of cervical spine flexion-extension, lateral bending and axial rotation. RESULTS: Average length changes ranged from 0.01 to 0.13% (SD 0.01-0.21%) of initial length for all motions and locations studied, which in all cases was less than the accuracy of the transducers. CONCLUSION: It can thus be concluded that cervical spine motion does not induce significant strain of the cerebral dura mater. RELEVANCE: The present study does not support theories that are based on the transmission of strains from spinal to cranial dura mater. PMID- 12763435 TI - Biomechanical changes elicited by an anterior cruciate ligament deficiency during steady rate cycling. AB - OBJECTIVES: To identify any changes to lower limb biomechanics during steady rate cycling as a result of an anterior cruciate ligament deficiency. DESIGN: Comparative study in which healthy and anterior cruciate ligament injured individuals underwent biomechanical analysis during stationary cycling. BACKGROUND: Individuals with an anterior cruciate ligament deficiency often exhibit reductions in the magnitude of quadriceps muscle activity and subsequent knee joint extensor moments during walking. It is not known whether these compensations are present during cycling, an exercise frequently used to retrain anterior cruciate ligament injured individuals. METHODS: Ten healthy and 10 unilateral anterior cruciate ligament deficient individuals participated. All participants were required to cycle for approximately 30 s at each of six different cycling intensities while lower limb EMG, kinetics, and kinematics were collected bilaterally. Before riding, participants performed submaximal isometric contractions to generate normalizing data. RESULTS: In addition to reduced quadriceps activation and net knee joint extensor moments, the anterior cruciate ligament deficient limbs exhibited decreases in linear impulse of the resultant pedal force, knee joint flexor moments, hip and ankle extensor moments, and muscle activity from gluteus maximus. These decreases were counteracted by an increase in output from the anterior cruciate ligament intact limb. CONCLUSION: Anterior cruciate ligament injured individuals exhibited a limb attenuation strategy during cycling activities. RELEVANCE: This study reports lower limb kinetic and electromyographic data from anterior cruciate ligament deficient individuals during stationary cycling, and shows that these individuals exhibit a limb attenuation strategy on the very leg that is undergoing rehabilitation. PMID- 12763436 TI - In vivo and noninvasive six degrees of freedom patellar tracking during voluntary knee movement. AB - OBJECTIVE: The purpose of this study was to investigate in vivo and noninvasively patellar tracking in six degrees of freedom during voluntary knee extension and flexion. DESIGN: Patellar tracking was evaluated in vivo and noninvasively with corroboration using in vivo fluoroscopy and in vitro cadaver measurements. BACKGROUND: Patellofemoral pain is closely related to abnormal patellar tracking and malalignment. However, there is a lack of quantitative and convenient methods to evaluate six degrees of freedom in vivo patellar tracking, partly due to difficulty in evaluating 3-D patellar tracking noninvasively. METHODS: Six degrees of freedom patellar tracking was measured in vivo and noninvasively using a small clamp mounted onto the patella and an optoelectronic motion capture system in 18 knees of 12 healthy subjects during voluntary knee extension and flexion. RESULTS: The patella tracked systematically following a certain pattern during knee extension and flexion. Patellar tracking patterns during knee extension and flexion were not significantly different in the 18 knees tested. When the knee was voluntarily extended from 15 degrees flexion to full extension, the patella was extended 8 degrees, laterally tilted 2 degrees, and shifted 3 mm laterally and 10 mm proximally. The results were consistent with previous in vitro and in vivo studies. CONCLUSION: Six degrees of freedom patellar tracking can be evaluated in vivo and noninvasively within the range of 20 degrees flexion to full knee extension. RELEVANCE: The study provided us quantitative six degrees of freedom information about patellar tracking during knee flexion/extension, which can be used to investigate patellar tracking in vivo and noninvasively in both healthy subjects and patients with patellofemoral disorder and patellar malalignment. PMID- 12763437 TI - Mobile and fixed bearing total knee prosthesis functional comparison during stair climbing. AB - OBJECTIVE: The purpose of this investigation is to determine the functional performance of the mobile bearing total knee replacement prosthesis as compared to the fixed bearing type total knee replacement prosthesis. DESIGN: Kinematics, kinetics, and electromyography data were gained from 10 patients with mobile bearing and 10 patients with a fixed bearing posterior stabilized Insall Burstein II total knee replacement during ascending and descending stairs. A control group of 10 normal subjects, matched by sex and age, was also analysed. BACKGROUND: No significant biomechanical differences in patients with different total knee replacement designs have been reported from level-walking studies. Slightly better performance of posterior retaining with respect to cruciate sacrificing total knee replacement designs have been claimed from stair climbing studies. Only one study has been conducted regarding mobile versus fixed bearing total knee replacement assessed by gait analysis. This study did not show any biomechanical differences between the two groups. METHODS: Motion analysis was used to quantify the knee kinematics, kinetics, and electromyography (right and left longissimus dorsi, gluteus medius, rectus femoris, biceps femoris, semitendinosus, gastrocnemius and tibialis anterior muscles) during stair ascent and descent. RESULTS: The mobile bearing group demonstrated a reduced knee extensor moment during stair climbing and descending, and a reduced knee adductor moment during stair climbing. When ascending stairs, most of the mobile bearing patients show a peak knee flexion and a peak knee flexion moment at the late stance phase during the double support period. This kinematic and kinetic pattern is absent in normal subject. Both mobile bearing and fixed bearing groups showed abnormal electromyography patterns in both descending and ascending. CONCLUSIONS: During stair climbing, the mobile bearing design demonstrates a different kinematic pattern to the fixed bearing total knee replacement. Lower limb compensatory mechanisms seemed to be adopted particularly by the mobile bearing patients during ascending stairs. RELEVANCE: Total knee replacement patient with mobile bearing design can feel excessive femoro-tibial motion during daily living activities such as stair climbing and descending. Proprioceptive control of this tibio-femoral translation is needed as demonstrated by the lower limb compensatory mechanism. This data suggest that antero-posterior constraint structures (ligamentous or mechanical) are important to obtain reproducible knee kinematics. PMID- 12763438 TI - The upper body segmental movements during walking by young females. AB - OBJECTIVES: To characterise the physiological pattern of trunk and shoulders movements during walking and provide a reference for further studies on spine deformities. DESIGN: Implementation of a model for measuring spine and shoulder girdle movements during gait. Data collection on a population of eighteen, young, healthy, female subjects. BACKGROUND: The analysis of gait modifications in subjects with idiopathic scoliosis could offer an insight to better understand the functional relationship with the pathology. METHODS: Retroreflective markers were positioned on the main spine processes and acromions to be detected by a TV based motion analysis system. A model of kinematic computation was implemented and integrated in a previously developed protocol for multifactorial gait analysis. Movements in the main reference planes and in relation to pelvis were analysed. RESULTS: The trunk was on average bent forward by 3.4 degrees with respect to standing; of the two physiological curves in the sagittal plane only lordosis changed during walking; in the frontal plane, a dynamic spine deformation appeared, that was maximum at heel strike-early stance; the trunk was bent controlaterally of the foot on the ground, while the shoulders remained stable; in the horizontal plane, the shoulders rotated contralaterally to the pelvis. CONCLUSIONS: In our population all the segmental movements analysed were smaller than 5 degrees during gait, except the angle of proximal curvature in the frontal plane, shoulder rotation, and angle between shoulders and pelvis; all the measured angles were far below their possible ranges of motion. RELEVANCE: Quantitative data on upper body kinematics as a complement to gait analysis can help understanding movement disorders and compensation strategies in several pathologies. PMID- 12763439 TI - Influence of the diabetic neuropathy on the behavior of electromyographic and sensorial responses in treadmill gait. AB - OBJECTIVE: We describe and interpret self-cadence treadmill walking by neuropathic diabetic subjects under biomechanical and somatosensorial considerations. DESIGN: EMG variables during stance phase of neuropathic diabetic subjects were acquired and analyzed. We also evaluated sensorial and motor aspects of the feet and legs. METHODS: The experimental procedures are divided as follows: (a) determination of the sensitive cronaxie and pain tolerance in selected plantar areas, (b) determination and description of temporal aspects of EMG patterns of the vastus lateralis, tibialis anterior and lateral gastrocnemius of both sides during treadmill walking. We analyze and compare the results of the sensitive cronaxie, pain tolerance and the EMG parameters obtained by two experimental groups: diabetic neuropathic (n=20) and non-diabetic control subjects (n=20). RESULTS: The somatosensorial responses and pain tolerance threshold in the diabetic neuropathic group were significantly higher and considered far from the normal patterns. The EMG responses of the thigh and leg muscles in the diabetic neuropathic group were delayed if compared to the normal recruitment pattern, especially the tibialis anterior and vastus lateralis. CONCLUSIONS: These findings lead us to conclude that probably central and/or peripheral control mechanisms of the gait of neuropathic diabetic patients are altered due to somatosensorial and motor deficits. The mechanism of load reduction during walking was considered inefficient because of the activation delay of the vastus lateralis and tibialis anterior. We have concluded that the peripheral diabetic neuropathy damages not only somatosensorial and motor sources but also intrinsic mechanisms of motor control leading to alterations in the ankle efficiency in gait. This resulting distal inefficiency compromises some of the principal requirements for gait, such as progression and balance. RELEVANCE: This investigation is based on an innovating thematic approach involving the diabetic peripheral neuropathy. This innovation concerns the use of EMG and an instrumented treadmill in a clinical application to study and interpret the motor control during gait in neuropathic diabetic patients. PMID- 12763440 TI - Nonlinear dynamics indicates aging affects variability during gait. AB - OBJECTIVE: To investigate the nature of variability present in time series generated from gait parameters of two different age groups via a nonlinear analysis. DESIGN: Measures of nonlinear dynamics were used to compare kinematic parameters between elderly and young females. BACKGROUND: Aging may lead to changes in motor variability during walking, which may explain the large incidence of falls in the elderly. METHODS: Twenty females, 10 younger (20-37 yr) and 10 older (71-79 yr) walked on a treadmill for 30 consecutive gait cycles. Time series from selected kinematic parameters of the right lower extremity were analyzed using nonlinear dynamics. The largest Lyapunov exponent and the correlation dimension of all time series, and the largest Lyapunov exponent of the original time series surrogated were calculated. Standard deviations and coefficient of variations were also calculated for selected discrete points from each gait cycle. Independent t-tests were used for statistical comparisons. RESULTS: The Lyapunov exponents were found to be significantly different from their surrogate counterparts. This indicates that the fluctuations observed in the time series may reflect deterministic processes by the neuromuscular system. The elderly exhibited significantly larger Lyapunov exponents and correlation dimensions for all parameters evaluated indicating local instability. The linear measures indicated that the elderly demonstrated significantly higher variability. CONCLUSIONS: The nonlinear analysis revealed that fluctuations in the time series of certain gait parameters are not random but display a deterministic behavior. This behavior may degrade with physiologic aging resulting in local instability. RELEVANCE: Elderly show increased local instability or inability to compensate to the natural stride-to-stride variations present during locomotion. We hypothesized that this may be the one of the reasons for the increases in falling due to aging. Future efforts should attempt to evaluate this hypothesis by making comparisons to pathological subjects (i.e. elderly fallers), and examine the sensitivity and specificity of the nonlinear methods used in this study to aid clinical assessment. PMID- 12763441 TI - How much can a vibrational diagnostic tool reveal in total hip arthroplasty loosening? AB - OBJECTIVE: To investigate how much information a vibrational technique can reveal regarding the loosening of the femoral component of a total hip arthroplasty. DESIGN: Numerical modal analysis using finite element method and computer simulation. BACKGROUND: Existing work suggested that this technique is capable of revealing total hip arthroplasty loosening. However, the potentials of this technique are not fully studied yet. There is a need to establish the limitation of this technique. METHODS: Numerical modal analysis was conducted to obtain the natural frequencies and mode shapes of femoral models under free vibration. An absent interface portion was hypothesized at specified locations the data from modal analysis were then used in a computer simulation to visualize the vibration diagnosis. RESULTS: The effects of interface failure on the femoral component stiffness vary with the locations and sizes of failure. Nominal critical, reliable, and nondetectable levels of interface failure are found. CONCLUSION: When the size of failure is more than one-third of the stem length, this tool is able to provide a reliable diagnosis. The critical detectable interface failure size is one-fifth of the stem length. It was found that the motion at higher harmonics is the most sensitive to the interface failure. This tool is unlikely to differentiate the slight differences in natural frequencies that occur when the interface failure length is less than one-sixth of the stem length. It is likely to fail in detecting failures located at the stem central portion regardless of their length. RELEVANCE: A numerical study of the vibration tool is important prior to pursuing clinical trials of the tool and may guide the characterizations involved in the trials. PMID- 12763442 TI - Reliability of a new, hand-held device for assessing skeletal muscle stiffness. AB - OBJECTIVE: The aim of the study was to examine the test-retest reliability of the Myoton-2 myometer for measuring skeletal muscle viscoelastic stiffness. METHODS: Ten healthy volunteers took part. On day 1, the viscoelastic stiffness of the rectus femoris, vastus lateralis, biceps femoris, and gastrocnemius muscle (lateral and medial heads) was measured at rest using the Myoton-2 myometer. On day 2, the tests were repeated, and the rectus femoris was also examined during the maintenance of submaximal contractions of the quadriceps, and at a different resting muscle length. RESULTS: The myometer showed good to excellent test-retest reliability for all muscles (ICCs 0.80-0.93), except for the vastus lateralis (ICC 0.40). Viscoelastic stiffness showed a linear increase with increasing quadriceps' force output, and was higher in stretched than in shortened resting muscle. CONCLUSION: The Myoton-2 myometer is a reliable device for measuring the viscoelastic stiffness of resting muscle. Furthermore, viscoelastic stiffness showed the expected changes in response to increases in force output and muscle length, suggesting that the measurements were also valid. RELEVANCE: The results of this pilot study show that the Myoton-2 myometer is a simple, precise instrument for measuring muscle viscoelastic stiffness. If the findings can be confirmed in larger studies, further research should be carried out to examine its potential applications in the field of musculoskeletal medicine. PMID- 12763443 TI - Development and testing of a pelvic goniometer designed to measure pelvic tilt and hip flexion. AB - OBJECTIVE: To determine the reliability and validity of a pelvic goniometer designed to measure the pelvic tilt and hip flexion during seated posture. BACKGROUND: Assessment of the seated posture requires measurement of the pelvis and hip. Determining accurate pelvic tilt and hip flexion angles during sitting is often difficult using standard techniques. A pelvic goniometer has been designed to measure pelvic tilt and hip flexion angle of persons in a seated posture. METHODS: VALIDATION of the pelvic goniometer was done radiographically. Ten male volunteers sat in three postures--erect, forward or anterior tilt, and posterior tilt. Pelvic tilt and hip angle were recorded using radiographs and the pelvic goniometer. Reliability of pelvic and conventional goniometers was done using seated nondisabled subjects with physical therapists performing measurements. RESULTS: VALIDATION: the average differences and correlation between the pelvic goniometer and radiographic measures were as follows--pelvic tilt: -4.9 degrees, 0.93; hip angle 1.2 degrees, 0.81. Reliability: average range of hip angle across three measures was about 3 degrees for both goniometers. CONCLUSIONS: The data indicate that the pelvic goniometer has utility in measuring pelvic tilt and hip angle, especially within the seated posture. Because it measures both pelvic tilt and hip angle, the pelvic goniometer has an advantage over conventional goniometers that only measure the latter. RELEVANCE: A valid and reliable tool that measures pelvic tilt and hip angle of persons in a seated posture is needed for clinical research and practice. Its applications include wheelchair seating evaluations and ergonomic assessments of seated workers. PMID- 12763444 TI - Characterisation and quantification of angiogenesis in beta-tricalcium phosphate implants by immunohistochemistry and transmission electron microscopy. AB - The aim of this study is the histological characterisation of angiogenesis in a macroporous biomaterial with quantification techniques used in oncology. Porous tricalcium phosphate implants were seated in the tibias of 12 rabbits. This work allows (1) morphological study with photonic microscopy, transmission electron microscopic and immunohistochemistry labelling for (2) quantification of vascularisation using anti-CD31 monoclonal antibody (3) quantification of proliferation using anti-PCNA polyclonal antibody (4) study of two angiogenic growth factors: VEGF and FGF-2. Quantification of angiogenesis revealed an outbreak kinetic with early vascular growth in first several days and a second growth phase after 4 weeks. This study reveals in macropores many isolated cells without adjacent vascular lumen, with endothelial phenotype. Expression of angiogenic growth factors reveals that all endothelial cells were VEGF-negative throughout the test period. FGF-2 expression by endothelial cells began 2 weeks post-implantation. Osteoblasts strongly expressed two markers throughout the test period. Furthermore, the procedure described here can be used to compare angiogenesis in different biomaterials or in the same biomaterial with the influence of macroporosities. PMID- 12763445 TI - Biomechanical and histomorphometric investigations on two morphologically differing titanium surfaces with and without fluorohydroxyapatite coating: an experimental study in sheep tibiae. AB - The influence of fluorohydroxyapatite (FHA) coating and surface roughness of Ti6Al4V implants on bone response was investigated. Uncoated and FHA-coated screws with lower (LR and LR+FHA; Ra: 5.7+/-0.2 microm) and higher (HR and HR+FHA; Ra: 21.8+/-0.9 microm) surface roughness, were inserted into the diaphyses of 8 sheep tibiae. Twelve weeks after implantation, extraction torque and bone-to-implant contact were evaluated. The smoothest surfaces showed an improved extraction torque and significant differences were observed between LR and HR (-24.6%, p<0.0005), LR and HR+FHA (-30.7%, p<0.0005), LR+FHA and HR ( 17.4%, p<0.005), and LR+FHA and HR+FHA (-24.0%, p<0.005). The bone-to-implant contact data paralleled the biomechanical data: the smoother the surface, the greater the bone-to-implant contact. Significant (p<0.0005) decreases in bone-to implant contact were observed between LR+FHA and HR (-24.2%), and between LR+FHA and HR+FHA (-29.2%). The current findings suggest that LR surfaces significantly improve the osteointegration rate of implanted cortical screws independently of the FHA coating. PMID- 12763446 TI - Effect of consolidation on adhesive and abrasive wear of ultra high molecular weight polyethylene. AB - Total hip replacement (THR) is widely performed to recover hip joint functions lost by trauma or disease and to relieve pain. The major cause of failure in THR is the wear of the ultra high molecular weight polyethylene (UHMWPE) component. The dominant wear mechanism in THR occurs through adhesion and abrasion. While poor consolidation of UHMWPE is known to increase the incidence of a different damage mode, delamination, which is the dominant wear mechanism in tibial inserts but uncommon in THR, the effect of consolidation on adhesive and abrasive wear of UHMWPE is not clear. In this study UHMWPE resin was subjected to hot isostatic pressing under a pressure of 138MPa at different temperatures (210 degrees C, 250 degrees C, and 300 degrees C) to achieve varying degrees of consolidation. The extent of consolidation was determined by optical microscopy using thin sections, and by scanning electron microscopy using cryofractured and solvent etched specimens. Wear behavior of the samples with varying degree of consolidation was determined using a bi-directional pin-on-disc machine simulating conditions in a hip joint. Increasing the processing temperature decreased the incidence of fusion defects and particle boundaries reflecting the powder flakes of the virgin resin, improving the consolidation. However, the bi-directional pin-on-disc wear rate did not change with the processing temperature, indicating that adhesive and abrasive wear is independent of the extent of consolidation in the range of parameters studied here. PMID- 12763447 TI - In vivo bone and soft tissue response to injectable, biodegradable oligo(poly(ethylene glycol) fumarate) hydrogels. AB - This study was designed to assess in vivo bone and soft tissue behavior of novel oligo(poly(ethylene glycol) fumarate) (OPF) hydrogels using a rabbit model. In vitro degradation of the OPF hydrogels was also investigated in order to compare with in vivo characteristics. Four groups of OPF hydrogel implants were synthesized by alternation of crosslinking density, poly(ethylene glycol) (PEG) block length of OPF, and cell-binding peptide content. The in vitro degradation rate of OPF hydrogels increased with decreasing crosslinking density of hydrogels, which was characterized by measuring weight loss and swelling ratio of hydrogels and medium pH change. Examination of histological sections of the subcutaneous and cranial implants showed that an uniform thin circumferential fibrous capsule was formed around the OPF hydrogel implants. Quantitative evaluation of the tissue response revealed that no statistical difference existed in capsule quality or thickness between implant groups, implantation sites or implantation times. At 4 weeks, there was a very limited number of inflammatory and multinuclear cells at the implant-fibrous capsule interface for all implants. However, at 12 weeks, OPF hydrogels with PEG block length of number average molecular weight 6090+/-90 showed extensive surface erosion and superficial fragmentation that was surrounded by a number of inflammatory cells, while OPF hydrogels with PEG block length of number average molecular weight 930+/-10 elicited minimal degradation. Constant fibrous capsule layers and number of inflammatory cells were observed regardless of the incorporation of cell-binding peptide and crosslinking density of OPF hydrogels with PEG block length of number average molecular weight 930+/-90. These results confirm that the degradation of implants can be controlled by tailoring the macromolecular structure of OPF hydrogels. Additionally, histological evaluation of implants proved that the OPF hydrogel is a promising material for biodegradable scaffolds in tissue engineering. PMID- 12763448 TI - Crosslinked collagen/chitosan matrix for artificial livers. AB - Matrices composed of collagen and chitosan may create an appropriate environment for the regeneration of livers. In this study, we have prepared, characterized and evaluated a new collagen/chitosan matrix (CCM). The CCM was made by using crosslinking agent 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC) in N hydroxysuccinimide (NHS) and a 2-morpholinoethane sulfonic acid (MES) buffer system. The chemical characteristics were evaluated by Fourier-transformed infrared (FTIR) spectroscopy and X-ray photoelectron spectroscopy (XPS). The mechanical strength was measured by tensile tests. The platelet deposition and hepatocyte culture experiments show that CCM has excellent blood and cell compatibility. The results suggest that the CCM is a promising candidate matrix for implantable bioartificial livers. PMID- 12763449 TI - Formation of Propionibacterium acnes biofilms on orthopaedic biomaterials and their susceptibility to antimicrobials. AB - Failure to treat and eradicate prosthetic hip infection with systemic antibiotic regimens is usually due to the fact that the infection is associated with biofilm formation and that bacterial cells growing within a biofilm exhibit increased resistance to antimicrobial agents. In this in vitro study, we investigated the susceptibility of prosthetic hip Propionibacterium acnes and Staphylococcus spp. isolates growing within biofilms on polymethylmethacrylate (PMMA) bone cement to a range of antibiotics. All P. acnes isolates in the biofilm mode of growth demonstrated considerably greater resistance to cefamandole, ciprofloxacin and vancomycin. In contrast, only four of the eight P. acnes isolates demonstrated an increase in resistance to gentamicin. All ten Staphylococcus spp. isolates in the biofilm mode of growth exhibited large increases in resistance to gentamicin and cefamandole with eight of the ten isolates also exhibiting an increase in resistance to vancomycin. However, only three of the ten Staphylococcus spp. isolates exhibited an increase in resistance to ciprofloxacin. Biofilms were also formed on three different titanium alloys and on PMMA bone cement using P. acnes, Staphylococcus epidermidis and Staphylococcus aureus strains to determine if the underlying biomaterial surface had an effect on biofilm formation and the antimicrobial susceptibility of the bacteria growing within biofilms. Although differences in the rate at which the three strains adhered to the different biomaterials were apparent, no differences in biofilm antibiotic resistance between the biomaterials were observed. In the light of these results, it is important that the efficacy of other antibiotics against P. acnes and Staphylococcus spp. prosthetic hip isolates growing within biofilms on orthopaedic biomaterials be determined to ensure optimal treatment of orthopaedic implant infection. PMID- 12763450 TI - Establishing the protein MIA (melanoma inhibitory activity) as a marker for chondrocyte differentiation. AB - Melanoma inhibitory activity (MIA), also referred to as cartilage derived retinoic acid-sensitive protein (CD-RAP), is detected physiologically in cartilage tissue and pathologically in malignant melanomas. To measure MIA/CD-RAP quantitatively we developed a sensitive ELISA system. Recently, we described diagnostic applications of the MIA-ELISA in patients with cartilage diseases. The study described herein was performed to determine whether there is any relation between MIA/CD-RAP levels and the degree of chondrocyte differentiation in tissue culture and to analyse whether MIA/CD-RAP may serve as a useful marker to control chondrocyte differentiation in in vitro tissue engineering. Our data provide evidence that measuring MIA in tissue culture supernatant by a quantitative ELISA can be used as a marker for differentiated chondrocytes. PMID- 12763452 TI - Preparation of poly(lactic acid) composites containing calcium carbonate (vaterite). AB - A new type of ceramic-polymer biomaterial having excellent apatite-forming ability in simulated body fluid was prepared by hot-pressing a mixture of poly(-L lactic acid) (PLA) and calcium carbonate (vaterite). After PLA dissolved in methylene chloride was mixed with calcium carbonate consisting of vaterite, the mixture was dried completely and subsequently hot-pressed uniaxially under a pressure of 40 MPa at 180 degrees C. When 30 wt% vaterite was introduced, the modulus of elasticity was effectively improved by 3.5-6 GPa, which was about twice higher than the modulus of PLA. The composite showed no brittle fracture behavior and a comparably high bending strength of approximately 50 MPa. The composite containing 30 wt% vaterite formed a 5-15-microm-thick bonelike apatite layer on its surface after soaking in SBF at 37 degrees C even for 1-3d. PMID- 12763451 TI - Serum protein adsorption and platelet adhesion on pluronic-adsorbed polysulfone membranes. AB - We examined plasma protein adsorption and platelet adhesion to polysulfone (PSf) flat membranes coated with Pluronic with varying polyethylene oxide (PEO) block length. Adsorption of albumin, globulin and fibrinogen to Pluronic-coated PSf membranes was independent of plasma dilution when concentrations of human blood plasma above 20% were applied. Increasing coating concentrations of aqueous Pluronic solution resulted in decreased protein adsorption by the PSf membranes. Pluronic F68, which was more hydrophilic than Pluronic L62 or L64 and had 80% of PEO content, was the most effective at suppressing the adsorption of plasma proteins and platelet adhesion to PSf membranes. We developed a mixed protein solution containing human albumin, gamma-globulin and fibrinogen to attempt to mimic the competitive and cooperative binding effects found in plasma. Fibrinogen adsorption from plasma could be recapitulated by the mixed protein solution. The number of platelets adhering to the PSf membranes decreased as the coating concentration of Pluronic solution was increased, and platelet adhesion decreased in parallel with fibrinogen adsorption. These results suggest that the bioinert property of PEO segments in the Pluronic, which is ascribed to their high flexibility in aqueous media, suppresses the adsorption of plasma proteins and platelets to the Pluronic-coated PSf membranes. PMID- 12763453 TI - Cross-linked type I and type II collagenous matrices for the repair of full thickness articular cartilage defects--a study in rabbits. AB - The physico-chemical properties of collagenous matrices may determine the tissue response after insertion into full-thickness articular cartilage defects. In this study, cross-linked type I and type II collagen matrices, with and without attached chondroitin sulfate, were implanted into full-thickness defects in the femoral trochlea of adolescent rabbits. The tissue response was evaluated 4 and 12 weeks after implantation by general histology and two semi-quantitative histological grading systems. Four weeks after implantation, type I collagenous matrices were completely filled with cartilage-like tissue. By contrast, type II collagenous matrices revealed predominantly cartilaginous tissue only at the superficial zone and at the interface of the matrix with the subchondral bone, leaving large areas of the matrix devoid of tissue. Attachment of chondroitin sulfate appeared to promote cellular ingrowth and cartilaginous tissue formation in both types of collagen matrices. Twelve weeks after implantation, the differences between the matrices were less pronounced. The deep parts of the subchondral defects were largely replaced by new bone with a concomitant degradation of the matrices. The original cartilage contours in defects with type I collagen-based matrices were repaired with fibro-cartilaginous tissue. Defects containing type II matrices showed an increase in the amount of superficial cartilage-like tissue. The original contour, however, was not completely restored in all animals, occasionally leaving a central depression or fissure. It is concluded that different types of collagen matrices induce different tissue responses in full-thickness articular cartilage defects. Type I collagen-based matrices are superior to guide progenitor cells from a subchondral origin into the defect. In type II collagen-based matrices cell migration is less, but invading cells are directed into a chondrocyte phenotype. Based on these observations it is suggested that a composite matrix consisting of a deep layer of type I collagen and a more superficial layer of type II collagen may be the matrix of choice for cartilage regeneration. PMID- 12763455 TI - Porous ZrO2 bone scaffold coated with hydroxyapatite with fluorapatite intermediate layer. AB - Highly porous zirconia (ZrO(2)) bone scaffolds, fabricated by a replication technique using polymeric sponge, were coated with hydroxyapatite (HA). To prevent the chemical reactions between ZrO(2) and HA, an intermediate fluorapatite (FA) layer was introduced. The strength of the porous ZrO(2) was higher than that of pure HA by a factor of 7, suggesting the feasibility of ZrO(2) porous scaffolds as load-bearing part applications. The coated HA/FA layer, with a thickness of about 30 microm, was firmly adhered to the ZrO(2) body with a bonding strength of 22MPa. The osteoblast-like cells were attached and spread well on the coating layer throughout the porous scaffolds. The alkaline phosphatase activity of the proliferated cells on the HA/FA coated ZrO(2) was comparable to that on pure HA and higher than that on pure ZrO(2). PMID- 12763454 TI - Tissue-specific gene expression in chondrocytes grown on three-dimensional hyaluronic acid scaffolds. AB - The re-differentiation capacities of human articular and chick embryo sternal chondrocytes were evaluated by culture on HYAFF-11 and its sulphate derivative, HYAFF-11-S, polymers derived from the benzyl esterification of hyaluronate. Initial results showed that the HYAFF-11-S material promoted the highest rate of chondrocyte proliferation. RNA isolated from human and chick embryo chondrocytes cultured in Petri dishes, HYAFF-11 or HYAFF-11-S were subjected to semi quantitative RT-PCR analyses. Human collagen types I, II, X, human Sox9 and aggrecan, chick collagen types I, II, IX and X were analysed. Results showed that human collagen type II mRNA expression was upregulated on HYAFF-11 biomaterials. In particular, a high level of collagen type IIB expression was associated with three-dimensional culture conditions, and the HYAFF-11 material was the most supportive for human collagen type X mRNA expression. Human Sox9 mRNA levels were constantly maintained in monolayer cell culture conditions over a period of 21 days, while these were upregulated when chondrocytes were cultured on HYAFF-11 and HYAFF-11S. Furthermore, chick collagen type IIA and IIB mRNA expression was detected after only 7 days of HYAFF-11 culture. Chick collagen type IX mRNA expression decreased in scaffold cultures over time. Histochemical staining performed in engineered cartilage revealed the presence of a de novo synthesized glycosaminoglycan-rich extracellular matrix; immunohistochemistry confirmed the deposition of collagen type II. This study showed that the three-dimensional HYAFF-11 culture system is both an effective chondrocyte delivery system for the treatment of articular cartilage defects, and an excellent in vitro model for studying cartilage differentiation. PMID- 12763456 TI - The chitosan prepared from crab tendons: II. The chitosan/apatite composites and their application to nerve regeneration. AB - The chitosan tubes derived from crab tendons form a hollow tube structure, which is useful for nerve regeneration. However, in order to use the chitosan tubes effectively for nerve regeneration, there remain two problems to be solved. First, the mechanical strength of the tubes is quite high along the longitudinal axis, but is somewhat low for a pressure from side. Second, the chitosan tube walls swell to reduce the inner space of the tubes in vivo. These two problems limit the clinical use of the chitosan tubes. In this study, to solve the problems, apatite was made to react with the chitosan tubes to enhance the mechanical strength of the tube walls. Transmission electron microscopy showed that apatite crystals were formed in the walls of the chitosan tubes. The c-axis of the crystals aligned well in parallel with chitosan molecules. These results indicate that the apatite crystals grow in the tubes starting from the nucleation sites of the chitosan molecules, probably by forming complexes with amino groups of chitosan and calcium ions. Further, the tubes were thermally annealed at 120 degrees C to prevent from swelling, and simultaneously formed into a triangular shape to enhance the stabilization of the tube structure. By these treatments, the hollow tubes could keep their shape even in vivo after implantation. Animal tests using SD rats further showed that the chitosan tubes effectively induced the regeneration of nerve tissue, and were gradually degraded and absorbed in vivo. PMID- 12763457 TI - Preparation of porous hydroxyapatite scaffolds by combination of the gel-casting and polymer sponge methods. AB - A new technique of combining the gel-casting and polymer sponge methods is introduced in this study to prepare macroporous hydroxyapatite scaffolds, which provides a better control over the microstructures of scaffolds and enhances their mechanical properties. With this technique, we were able to produce scaffolds with mechanical and structural properties that cannot be attained by either the polymer sponge or gel-casting method. The scaffolds prepared have an open, uniform and interconnected porous structure with a pore size of 200-400 microm. A compressive yield strength of approximately 5 MPa equivalent to that of cancellous bone and a compressive modulus of approximately 8 GPa similar to that of cortical bone were achieved. The pore morphology, size, and distribution of the scaffolds were characterized using a scanning electron microscope. X-ray diffraction and Fourier transform infrared spectroscopy were used to determine the crystal structure and chemical composition of scaffolds, respectively. Scaffolds with desired porosity, pore size, and geometry can be prepared by using polymer sponges of appropriate structures. PMID- 12763458 TI - Enhancing effect of poly(L-lactide) on the differentiation of mouse osteoblast like MC3T3-E1 cells. AB - Poly(L-lactide) (PLLA) has bioabsorbability and biocompatibility, and it is used as biodegradable screws, pins and plates for internal bone fixation. The purpose of this study was to clarify the effects of low molecular weight (Mw) PLLA on the proliferation and differentiation of mouse osteoblast-like MC3T3-E1 cells. MC3T3 E1 cells were cultured with the concentration of 5-50 microg/ml of PLLA with weight average Mw of 5000 (PLLA-5k) and 10,000 (PLLA-10k) for 2 weeks using the micromass culture. Both PLLAs did not affect the proliferation of MC3T3-E1 cells. However, the calcifications of MC3T3-E1 cells were stimulated with increasing the concentration of the PLLAs. Then PLLA-5k increased the calcification of MC3T3-E1 cells more than PLLA-10k. Additionally, both PLLAs increased the alkaline phosphatase (ALP) activity and calcium content of MC3T3-E1 cells up to the similar level to the calcification. These results indicated that low Mw PLLA enhanced the differentiation of MC3T3-E1 cells with no effect on the proliferation. Moreover, it was suggested that the increase of the ALP activity was a key step to stimulate the calcification of MC3T3-E1 cells. The osteoconductivity of implanted PLLA would be based on the enhancing effect of low Mw PLLA on the differentiation of the osteoblasts. PMID- 12763459 TI - Biocompatibility of implantable synthetic polymeric drug carriers: focus on brain biocompatibility. AB - Numerous polymeric biomaterials are implanted each year in human bodies. Among them, drug delivery devices are potent novel powerful therapeutics for diseases which lack efficient treatments. Controlled release systems are in direct and sustained contact with the tissues, and some of them degrade in situ. Thus, both the material itself and its degradation products must be devoid of toxicity. The knowledge and understanding of the criteria and mechanisms determining the biocompatibility of biomaterials are therefore of great importance. The classical tissue response to a foreign material leads to the encapsulation of the implant, which may impair the drug diffusion in the surrounding tissue and/or cause implant failure. This tissue response depends on different factors, especially on the implantation site. Indeed, several organs possess a particular immunological status, which may reduce the inflammatory and immune reactions. Among them, the central nervous system is of particular interest, since many pathologies still need curative treatments. This review describes the classical foreign body reaction and exposes the particularities of the central nervous system response. The recent in vivo biocompatibility studies of implanted synthetic polymeric drug carriers are summarized in order to illustrate the behavior of different classes of polymers and the methodologies used to evaluate their tolerance. PMID- 12763460 TI - Pectin-based systems for colon-specific drug delivery via oral route. AB - Pectin-derived matrices are now being examined and tested for controlled drug delivery. Pectin is intact in the upper gastrointestinal tract and degraded by colonic microflora. The composition of this microflora remains relatively consistent across a diverse human population. Thus, pectin-derived drug carriers provide promising potential for colon-specific drug delivery. This paper reviews recent developments in pectin-derived formulations. Subjects reviewed include gelation of pectin, calcium cross-linked pectinate, composites of pectin and other polymers, technologies to fabricate pectin into useful drug delivery vehicles, and methods to evaluate release kinetics of incorporated drugs. This article discusses advantages, limitations, and possible future developments in pectin-based formulations with particular emphasis on the field of colon-specific drug delivery. PMID- 12763461 TI - The effect of orthodontic bonding materials on dental plaque accumulation and composition in vitro. AB - The aim of this study was to investigate the accumulation and composition of microcosm dental plaque on different orthodontic bonding materials using an in vitro model. Microcosm plaques were grown on discs of a range of bonding materials in a constant depth film fermentor. The biofilms were derived from human saliva and supplied with artificial saliva as a source of nutrients. The number of viable bacteria in the biofilms was determined and the streptococci present were identified to species level. The results showed that there was no significant difference in bacterial accumulation between different bonding materials, however, biofilms grown on materials which were fluoride releasing, did not contain Streptococcus mutans. This in vitro study has shown that the use of fluoride-releasing bonding materials may support the growth of supragingival plaque, which does not contain S. mutans. PMID- 12763462 TI - Long-term radiographic changes in cemented total hip arthroplasty with six designs of femoral components. AB - Measurements were made from annual follow-up radiographs, obtained over 27.6 years, of 860 cemented total hip arthroplasties implanted by one surgeon. Femoral components were made of stainless steel or titanium alloy, were non-modular, and were all fixed with cement, and acetabular cups were all-polyethylene and were fixed with cement. Radiographic outcome was correlated with the shape and material of the femoral component. Specifically, throughout the follow-up, stems made of titanium alloy were at greater risk of developing bone-cement radiolucent lines than those made of stainless steel, the difference ranging from approximately 10-50 percent at 2-10 years of follow-up. Similarly, titanium alloy stems were at greater risk of developing endosteal scalloping, indicating osteolytic lesions. Among the stainless steel Charnley cobra and straight-narrow Charnley stems, none developed cement fracture, only one became radiographically loose and one developed endosteal scalloping. The differences in the risk of developing radiolucent lines, cement fracture and progressive loosening among these stems were correlated with the relative rigidity of the femoral stems, and were generally consistent with the predictions made heretofore using finite element models, although differences in stem surface finish and femoral ball size and material could have also influenced the results. PMID- 12763463 TI - Europium-doped bioapatite: a new photostable biological probe, internalizable by human cells. AB - The authors prepared at low temperatures (37 degrees C) a novel inorganic bioprobe. It consisted of mineral nanoparticles of apatitic tricalcium phosphate doped with europium, of size, structure and composition close to those of the mineral part of calcified tissues. In contrast to organic probes which degrade rapidly (photobleaching), the red luminescence of the new probe is photostable. Moreover, this luminescence can be obtained under visible irradiation, which makes it suitable for prolonged examination of live cells. Human pancreatic epithelial cells in culture were incubated with these particles and their internalization was observed by laser scanning confocal microscopy. Transmission electron microscopy and electron microdiffraction analysis confirmed that the particles were internalized retaining their original apatitic structure. This probe may thus be of value for biovectorization. PMID- 12763465 TI - Perspectives on epilepsy in people with intellectual disabilities: comparison of family carer, staff carer and clinician score profiles on the Glasgow Epilepsy Outcome Scale (GEOS). AB - PURPOSE: Clinical practice with people with intellectual disability relies heavily upon caregiver report. Crucially, the carer's perspective may depend upon his or her relationship to the patient. We investigated similarities and differences within and between family and paid carers in their reports on the Glasgow Epilepsy Outcome Scale (GEOS), an instrument that quantifies concerns about epilepsy in this population [Epilepsia 42 (2001) 1043]. METHODS: GEOS forms were available on 186 patients (108 males; mean age 39 years) across 384 primary respondents (141 staff, 83 family, 160 clinicians) and independently completed secondary respondents (67 staff, 36 family). Data were analysed to consider levels of concern as rated bv staff carers, family members and clinicians, and also to consider inter-rater agreement on the concerns raised. RESULTS: There were significant differences in the magnitude of concern on each sub-scale [concerns about seizures, treatment, caring and social impact; range of F(2,171)=9.5-64.7; all P<0.0001]. Post hoc testing revealed that family members scored all sub-scales more highly than staff carers or clinicians, and that staff carers scored more highly than clinicians on all but one sub-scale. Inter-rater agreement between family members was considerably higher (range of r=0.69-0.91) than between staff carers (r=0.30-0.47) across the GEOS sub-scales. Association between staff and family ratings was also modest (r< or = 0.39). CONCLUSIONS: It is preferable for the same staff member to complete each administration of the GEOS because of inter-staff variability in reporting of concerns. Families provide a consistent, but more extreme, picture and clinicians generally underestimate the concerns of direct caregivers. However, content of concerns varies relatively little across respondents. PMID- 12763466 TI - Family study of epilepsy in first degree relatives: data from the Italian Episcreen Study. AB - OBJECTIVE: To evaluate the family history of epilepsy in first degree relatives of probands with epilepsy. METHODS: A sample of 10787 patients with epilepsy with complete information about first degree relatives (parents, siblings and offspring) was selected from the database of the Episcreen Project, the largest Italian observational study on epilepsy. Family history was assessed by: (1) prevalence estimates of epilepsy among proband's relatives, (2) modified cumulative risks (MCR), adjusted using proband's age as censoring time in life tables, (3) standardised morbidity ratios (SMR), using a sub-group of symptomatic epilepsies as control group. RESULTS: Patients (9.1%) had a family history of epilepsy. The overall prevalence of epilepsy among first degree relatives was 2.6%. Idiopathic generalised epilepsies had the highest prevalence (5.3%). Cryptogenetic epilepsies had a lower prevalence (2.1%) than idiopathic epilepsies, but higher then symptomatic epilepsies (1.5%), both in generalised and focal forms (3.8% vs. 2.0% and 1.8% vs. 1.3%). A similar tendency was detected using MCR and SMR, with the higher values of risks/ratios for idiopathic and generalised epilepsies. Probands with idiopathic generalised epilepsies were highly concordant with respect to their relatives' type of epilepsy. Considering other strata factors, risks were higher in proband's epilepsies with an onset less then 14 years of age, while sex played no definite role in differentiating the family history. CONCLUSIONS: The Episcreen model permits a variety of stratification factors to measure family risk, including age at onset, epilepsy localisation and aetiology with a large sample of more than 10,000 probands and 1065/40,544 relatives affected and classified. PMID- 12763468 TI - Primary generalized epilepsy: a risk factor for seizures in labor and delivery? AB - PURPOSE: Women in the United States who have epilepsy give birth to about 20000 newborns every year. Because seizures during late gestation and delivery may seriously affect the fetus, and because primary generalized tonic-clonic (GTC) seizures may occur during labor and delivery in 1-2% of women with epilepsy, we attempted to define the rate, risks, and causes of seizures during labor and delivery. METHODS: To characterize seizures during labor and delivery, we retrospectively analyzed 89 consecutive pregnancies of women with epilepsy on antiepileptic drugs (AEDs). Six epileptologists in our group had treated these patients. We confirmed data and acquired new information by telephone for 83.1% of the pregnancies, and categorized the women as having primary generalized or partial epilepsy. Most of the patients (78%) were on monotherapy during pregnancy; 20% took two AEDs, and 3% took three AEDs during that period. RESULTS: Seizures during labor and delivery occurred in 4/32 (12.5%) patients with primary generalized epilepsy, but in none of the 57 women with partial epilepsy (P<0.05). None of the 38 patients with therapeutic AED levels before labor and delivery had seizures, compared to 3/37 (8.1%) of the subtherapeutic group. However, drug levels were taken at variable times in relation to delivery, limiting their value. Also, the levels sampled were both total and free levels; the latter would be more helpful to determine the adequacy of AED drug coverage. CONCLUSIONS: Maintaining therapeutic AED levels during the last trimester may help prevent seizures during labor and delivery, especially in women with generalized epilepsy. Women with epilepsy who had subtherapeutic AED levels and had been seizure-free may be at-risk for seizures during labor and delivery. Our sample was small, and a random sampling bias may have affected the results. PMID- 12763467 TI - A retrospective analysis of patients with febrile seizures followed by epilepsy. AB - This study was performed to investigate some clinical parameters of febrile seizures (FSs) in patients with epilepsy, testing any possible correlation between those parameters and the type of subsequent epilepsy. One hundred and nine patients with epilepsy having a history of FSs were evaluated for age at onset of FSs, interval between first FS and first afebrile seizure, recurrence rate, type of FSs, incidence of febrile status, family history for epilepsy and for FSs and the neurological status of the patient. The epilepsy that developed subsequently, were classified as generalised versus partial and also according to their syndromic subgroups. In temporal lobe epilepsy with mesial temporal sclerosis (TLE-MTS), statistical analyses revealed a younger age at onset of FSs, and a high incidence of episodes of febrile status and of complex FSs. Clinical characteristics of FSs followed by partial epilepsies were younger age at onset, presence of focal features and of febrile status, longer interval between the first FS and the first afebrile seizure, and a high incidence of FSs in the family history. In generalised epilepsies, however, a shorter interval between the first FS and the first afebrile seizure, a high incidence of single FS and of a family history of epilepsy were predominating characteristics. Results suggest that certain features of FSs may be predictive of a particular type of subsequent epilepsy. PMID- 12763469 TI - Magnetoencephalographic study of rhythmic mid-temporal discharges in non epileptic and epileptic patients. AB - To evaluate the source location and clinical significance of rhythmic mid temporal theta discharges (RMTD) by MEG in non-epileptic and epileptic patients, we conducted simultaneous MEG and EEG recordings with a whole-scalp 306-channel neuromagnetometer in three patients: one with right temporal lobe epilepsy (TLE), one with right frontal lobe epilepsy (FLE), and one with tension headache. We visually detected the RMTD activity and interictal spikes, and then localised their generators by MEG source modelling. We repeated MEG measurement 3 months after right anterior temporal lobectomy (ATL) in the TLE patient; 3 months after anticonvulsant medication in the FLE patient. In epileptic patients, RMTD activities were found during drowsiness over the left temporal channels of both MEG and EEG recordings, and their generators were localised to the left posterior inferior temporal region. In the patient with tension headache, RMTD was localised in the right inferior temporal area. When the epileptic patients became seizure free with disappearance of epileptic spikes, RMTD was still found over the left temporal channels. Besides, some bursts of RMTD appeared also in the right temporal channels in our TLE patient after ATL. Our results indicate that the source of RMTD activity is located in the fissural cortex of the posterior inferior temporal region. As a physiologic rhythm related to dampened vigilance, RMTD has no direct relation to epileptogenic activity. PMID- 12763470 TI - Clobazam monotherapy in drug naive adult patients with epilepsy. AB - PURPOSE: Evaluation of the efficacy and side effects profile of Clobazam in a 24 week open-labelled trial involving 26 cases of drug naive adult patients with epilepsy. METHODS: The study was an open labelled unicentre trial in which only drug naive cases with epilepsy were included. A total of 26 cases were recruited. One case was dropped because he did not complete the desired follow up. Seizure type and frequency were recorded and follow up was done at 4, 8, 12, 18 and 24 weeks after initiation of therapy. The change in seizure severity, the dose of Clobazam required and development of side effects were recorded. RESULTS: The seizure types included GTCS (n=16), complex partial seizures (n=4), focal motor seizures with secondary generalisation (n=3) and juvenile myoclonic epilepsy (n=2). Out of 25 patients, 16 (64%) became seizure free, while five (20%) had >50% reduction in their seizure frequency. Thus, these 21 patients (84%) were considered to be well controlled. The commonest side effect seen was sedation, which was noted in 4 of the 25 patients (16%). However, in none of these four patients sedation was significant enough to warrant stoppage of therapy. Weight gain, gait ataxia, loss of short-term memory and breakthrough seizures were noted in one patient each. CONCLUSIONS: The efficacy of Clobazam coupled with the lack of significant side effects noted in our study makes it merit consideration as monotherapy in adult patients with epilepsy. PMID- 12763471 TI - Seizures and therapy in adolescents with uncomplicated epilepsy. AB - PURPOSE: This study aimed to describe seizures and their therapy among Swedish adolescents, aged 13-22, with active but uncomplicated epilepsy. METHOD: The adolescents answered questionnaires (158/193). Data were also obtained from medical records. RESULTS: Epileptic seizure types could be specified in 92.1% of the cases. Predominant types were Primary Generalised Tonic-Clonic Seizures and Partial Complex Seizures with Secondary Generalisation. Clinical diagnoses by physicians were unspecified in 25.8%. Ninety percent were on antiepileptic drugs (AEDs), most commonly valproate and carbamazepine. New AEDs were used in 9.3% of the cases and polytherapy in 13.9%. More than 40% of the respondents had seizures despite AED treatment. Side effects of AEDs were experienced by 61%, most commonly tiredness, concentration difficulties and headache. Patients on polytherapy experienced significantly more side effects. The choice of a new AED over a traditional one was not related to seizure type or seizure control. CONCLUSIONS: Many adolescents had persistent seizures despite treatment at a specialist regional epilepsy centre. This, plus the high reported rate of side effects of AED treatment, suggests that treatment is not optimal for the group studied. As traditional AEDs strongly dominated treatment possibly newly marketed AEDs are underused in this group. PMID- 12763472 TI - From swelling to sclerosis: acute change in mesial hippocampus after prolonged febrile seizure. AB - Mesial temporal sclerosis (MTS) has been linked to prolonged febrile seizures. The sequence of changes in the temporal lobe/hippocampus following prolonged febrile seizures and status epilepticus is beginning to be elucidated. We obtained repeated magnetic resonance imaging (MRI) volumetric analysis of the hippocampi in a 23-month-old boy after a prolonged focal febrile seizure. Three days after a prolonged left focal febrile seizure, brain MRI showed increased T2 weighted signal and increased volume (swelling) of the right hippocampus. Repeat MRI 2 months later demonstrated sclerosis of the right hippocampus. Review of the literature shows four other children with prolonged focal seizures associated with the MRI sequence of temporal lobe swelling followed by sclerosis. All had left focal seizures followed by right MTS. Our patient demonstrates a shorter interval for the radiologic development of hippocampal sclerosis compared to other reports. PMID- 12763473 TI - Pilomotor seizures in frontal lobe epilepsy: case report. AB - We report a 27-year-old man with pilomotor seizures originating in the right frontal lobe. Subtracted ictal SPECT coregistered with MRI showed multifocal hyperperfused areas in the anterior medial frontal area, mainly involving the cingulate gyrus. Chronic electrocorticography with subdural electrode arrays during the piloerection demonstrated that the pilomotor seizures were originating in the anterior medial frontal region. After resection of the focus, the patient became seizure free. PMID- 12763474 TI - Complex partial status epilepticus of extratemporal origin in a patient with systemic lupus erythematosus. AB - The purpose of this case report is to describe the clinical, electroencephalographic and neuroimaging findings from a woman with systemic lupus erythematosus presenting with complex partial status epilepticus (CPSE) of neocortical temporo-parieto-occipital origin. The patient experienced complex visual hallucinations that initially were attributed to treatment with corticosteroids; however, an electroencephalogram (EEG) demonstrated the epileptic aetiology of her symptoms. CPSE should be considered as a possible cause of altered mental status in lupus. An urgent EEG is essential to make an accurate diagnosis. PMID- 12763475 TI - Advances in B-cell epitope analysis of autoantigens in connective tissue diseases. AB - The characterization of autoantibody specificities in rheumatic diseases is important in both diagnostic and basic research areas. Identification of the epitopes recognized by autoantibodies and their clinical and biological significance is not a trivial task. Epitopes may range in complexity from simple linear sequences of amino acids to complex quaternary structures. In addition to this structural complexity the frequency with which an autoantigen and its epitopes are recognized in a patient population may be useful in diagnosis, defining disease subgroups, and may offer information on disease prognosis. In this review recent advances in the epitope mapping of autoantigens in connective tissue diseases are discussed, with particular emphasis placed on the methodologies used to identify epitopes and the classification of the structural features of epitopes. To illustrate the identification of epitope structure, clinically relevant autoantigens, including CENP-A, PM/Scl-100, fibrillarin, filaggrin, Ro-52, and dsDNA, are discussed as examples of each type of epitope. PMID- 12763476 TI - Immune responses in breast cancer patients immunized with an anti-idiotype antibody mimicking NeuGc-containing gangliosides. AB - A phase I clinical trial was conducted in patients with stage III/IV breast cancer who were treated with the anti-idiotype mAb 1E10 specific to an Ab1 mAb able to react specifically with N-glycolyl-containing gangliosides and with antigens expressed on human melanoma and breast carcinoma cells. Patients were treated with 1 or 2 mg of aluminum hydroxide-precipitated 1E10 mAb every other week for six injections. Two patients at each dose were reimmunized 7-9 months after completing the induction phase. In hyperimmune sera from eight of the nine patients who received at least four doses of anti-Id vaccine preparations, strong specific responses were observed both against 1E10 mAb and NeuGc-GM3 ganglioside (Ab3 Id+Ag+). Nonclassical Ab1' antibodies (Id-Ag+) were also elicited by 1E10 mAb vaccine treatment. There were no differences between the two levels of dose tested in relation to toxicity and immunogenicity. No evidence of serious or unexpected effects was observed. PMID- 12763477 TI - Reconstitution of B cell function in murine models of immunodeficiency. AB - Murine models of immunodeficiency were used to evaluate strategies that might allow B cell engraftment in patients with X-linked agammaglobulinemia. Mice with defects in Btk or mu heavy chain were given 2.5 x 10(6) bone marrow cells from wild-type congenic donors. In the absence of any preparative regimen or immunosuppression, Btk-deficient mice on the CBA background developed normal concentrations of serum IgM and IgG3 by 12 weeks posttransplant. By contrast, mu heavy chain-deficient mice on the C57BL/6 background required some immunosuppression to achieve engraftment. Treatment of these mice with anti-T cell antibodies 2 and 4 days prior to transplant resulted in normal concentrations of serum immunoglobulins by 6 weeks posttransplant. These pretreated mice had only 10% of the normal number of splenic B cells and they had no evidence of donor T cell engraftment. These results suggest that myelotoxic drugs may not be needed to achieve B cell engraftment in B-cell-deficient subjects. PMID- 12763478 TI - Stem cell transplants for patients with X-linked agammaglobulinemia. AB - Six young patients with X-linked agammaglobulinemia and proven mutations in Btk were treated with cord blood or bone marrow transplants from HLA-matched siblings. Complete blood counts, serum chemistries, serum immunoglobulin concentrations, lymphocyte cell surface markers, and physical findings were evaluated at 3- to 5-day intervals for the first 2 weeks after transplant and then every 3 to 6 months. The first three patients were not given any preparative regimen or antirejection drugs and at 24 to 42 months posttransplant these patients have shown no benefit or harm related to the transplants. The second three patients were not given a preparative regimen but were treated with cyclosporine A (70 days) and mycophenolate mophetil (28 days) after transplant. Two of these patients have developed normal sized, nontender cervical lymph nodes 3 to 12 months after transplant but none of the three patients have shown an increase in serum IgM or an increase in the number of peripheral blood B cells. It is likely that successful engraftment will require more aggressive immunosupressive medications. PMID- 12763479 TI - Deficient activation and resistance to activation-induced apoptosis of CD8+ T cells is associated with defective peripheral tolerance in nonobese diabetic mice. AB - Activation-induced cell death (AICD) is a mechanism of homeostasis that limits the clonal expansion of autoreactive T cells and regulates central and peripheral tolerance. In nonobese diabetic (NOD) mice, defects in central and peripheral tolerance are associated with a proliferative hyporesponsiveness of thymocytes and peripheral T cells elicited upon TCR activation. We investigated whether these defects in tolerance induction and hyporesponsiveness of NOD T cells manifest in an altered susceptibility to TCR-induced AICD. TCR-activated NOD splenic CD4+ and CD8+ T cells are more resistant to AICD than control strain C57BL/6, BALB/c, and NOR T cells. NOR CD4+ but not CD8+ T cells are resistant to TCR-induced AICD. Whereas c-FLIP expression is reduced in activated T cells from control strains, it persists in activated NOD CD8+ T cells and is accompanied by diminished activity of caspase-3 and -8. IL-4 reduces this c-FLIP expression and increases caspase-3 and -8 activity in activated NOD CD8+ T cells. Moreover, IL-4 and CD28 costimulation restores the susceptibility of NOD CD8+ T cells to AICD, and this is associated with increased expression of CD25, CD95, CD95L, and TNFR2. Thus, deficient activation of CD8+ T cells and their greater resistance to TCR induced AICD may mediate defective peripheral tolerance and the development of T1D in NOD mice. PMID- 12763480 TI - Influenza vaccination and Guillain Barre syndrome. AB - Acute and severe Guillain Barre Syndrome (GBS) cases reported following influenza vaccine to the Vaccine Adverse Events Reporting System (VAERS) database from 1991 through 1999 were examined. Endotoxin concentrations were measured using the Limulus amebocyte lysate assay in influenza vaccines. There were a total of 382 cases of GBS reported to the VAERS database following influenza vaccination (male/female ratio, 1.2). The median onset of GBS following influenza vaccine was 12 days (interquartile range, 7 days to 21 days). There was an increased risk of acute GBS (relative risk, 4.3; 95% confidence interval, 3.0 to 6.4) and severe GBS (relative risk, 8.5; 95% confidence interval, 3.7 to 18.9) in comparison to an adult tetanus-diphtheria (Td) vaccine control group. There were maximums in the incidence of GBS following influenza vaccine that occurred approximately every third year (1993, 1996, and 1998) and statistically significant variation in the incidence of GBS among different influenza manufacturers. Influenza vaccines contained from a 125- to a 1250-fold increase in endotoxin concentrations in comparison to an adult Td vaccine control and endotoxin concentrations varied up to 10-fold among different lots and manufacturers of influenza vaccine. The biologic mechanism for GBS following influenza vaccine may involve the synergistic effects of endotoxin and vaccine-induced autoimmunity. There were minimal potential reporting biases in the data reported to the VAERS database in this study. Patients should make an informed consent decision on whether to take this optional vaccine based upon its safety and efficacy and physicians should vigilantly report GBS following influenza vaccination to the VAERS in the United States so that continued evaluation of the safety of influenza vaccine may be undertaken. PMID- 12763481 TI - Antigen-specific T cell response in infants after recombinant hepatitis B virus vaccination at birth: evaluation of T helper lymphocyte diversity. AB - Recombinant hepatitis B virus antigen (rHBsAg)-specific CD4+ T cell clones (TCC) were isolated and expanded from the peripheral blood of nine children vaccinated at birth against the hepatitis B (HB) virus. Four of them responded with protective antibody production (responders), three subjects were unable to produce detectable antibody levels even after revaccination (nonresponders), and two infants produced antibodies only after revaccination (slow responders). TCC were then characterized for their ability to produce cytokines known to be important for T cell expansion (interleukin-2, IL-2) and/or effector functions (IL-4, IFN-gamma, IL-10). Results demonstrated that the frequency of rHBsAg specific TCC in the samples of nonresponders was comparable to or higher than that in the samples of responders. Nevertheless, the majority of TCC obtained from responders or from slow responders before revaccination displayed the T helper 1 (T(H1))-dominant phenotype, while the majority of TCC obtained from nonresponders were nonpolarized T lymphocytes. After revaccination, the distribution of the different T(H) subsets in slow responders was heterogeneous. Overall, our present data suggest that an absence or delay in developing an rHBsAg-specific antibody response to vaccination is not associated with the capacity to generate an Ag-specific T cell response. However, compared to responders, nonresponding infants react to the rHBsAg vaccination with a reduced capacity to expand and differentiate toward polarized T(H) cells. PMID- 12763482 TI - The tetraspan protein EMP2 increases surface expression of class I major histocompatibility complex proteins and susceptibility to CTL-mediated cell death. AB - Dysregulation of class I major histocompatibility (MHC1) expression is an important mechanism of immunologic resistance for certain virus-infected or neoplastic cells. This study characterizes a new molecule affecting MHC1 expression and CTL cytotoxicity. Epithelial membrane protein 2 (EMP2) is a tetraspan protein recently identified for its role in suppressing B lymphoma tumorigenicity. The biochemistry of EMP2 suggests that it regulates the surface expression of certain membrane proteins, notably those destined for lipid raft microdomains. In this study, retroviral overexpression of EMP2 in target cells increased their susceptibility to CTL cytotoxicity. Conversely, down-expression of EMP2 using an EMP2-specific ribozyme rendered target cells CTL-resistant. EMP2 expression increased the surface levels of MHC1, CD54, and GM1 glycolipids. Biochemical fractionation indicated that these molecules reside with EMP2 in a lipid raft membrane compartment. Among MHC1 proteins, surface display of H-2D was particularly dependent on EMP2 expression, and blocking antibodies demonstrated that H-2D was critical for allogeneic CTL recognition. This study demonstrates an unexpected role for a tetraspan protein in CTL-mediated cell death and MHC1 surface trafficking. PMID- 12763483 TI - Gestational vascular complications. AB - Severe pregnancy complications, primarily severe pre-eclampsia, placental abruption, intrauterine growth restriction (IUGR) and intrauterine fetal death (IUFD) occur in about 1-5% of gestations. This rate is even higher in special medical situations. These pregnancy complications have been shown to increase maternal and fetal morbidity and mortality considerably. Severe pregnancy complications have also been shown to be associated with deficient uteroplacental circulation and are linked with intervillous and spiral vessel thrombosis. Moreover, it has been suggested that these complications could have their basis in a deficient trophoblast invasion in the uterine spiral arteries at a stage much earlier than the clinical manifestations become evident. In the last few years, evidence has accumulated to suggest that severe pregnancy complications could have a common thrombogenic basis associated with inherited and acquired thrombophilia. In this chapter we present a comprehensive update of the aetiology, pathophysiology, clinical manifestations, diagnosis and treatment of these severe pregnancy complications. PMID- 12763484 TI - Haemostatic changes in pregnancy. AB - Normal pregnancy is accompanied by changes in the coagulation and fibrinolytic systems. These include increases in a number of clotting factors (I, II, VII, VIII, IX and XII), a decrease in protein S levels and inhibition of fibrinolysis. As gestation progresses, there is also a significant fall in the activity of activated protein C, an important anticoagulant. While these physiological changes may be important for minimizing intrapartum blood loss, they entail an increased risk of thromboembolism during pregnancy and the post-partum period. PMID- 12763485 TI - Embryogenesis and gene targeting of coagulation factors in mice. AB - Genetic or acquired thrombophilia of the pregnant mother has been associated with the occurrence of gestational vascular disease and recurrent fetal loss and may contribute to the aetiology of pre-eclampsia. This chapter reviews insights into this link between thrombophilia and pregnancy complications that were gained from the study of genetically altered mice. These studies strongly support the notion of a cause-effect relationship between altered function of the thrombomodulin protein C pathway and adverse pregnancy outcome. Analysis of the mouse models highlights unique aspects of vascular structure and function at the feto-maternal interface, and exposes new biological functions of natural anticoagulant pathways in pregnancy. These roles are unrelated to the maintenance of vascular patency and may be mediated through specific signalling pathways activated by coagulation factors. Abnormal signalling by placental trophoblasts at the feto-maternal interface is suggested as a hitherto unrecognized mechanism that may underlie adverse pregnancy outcome associated with haemostatic disorders. PMID- 12763486 TI - Haemostatic mechanisms in human placenta. AB - The placenta is a unique organ with dual blood circulation functioning throughout fetal development. The architecture and functions of the placenta, where maternal blood flows into the intervillous space, present haemostatic problems, mainly the risk of haemorrhage. Placental trophoblasts express and produce coagulation components, participating not only in haemostasis but also in placental vascular development and differentiation. The expression of tissue factor, membrane phosphatidylserine and fibrin render the trophoblasts pro-coagulant, thus compromising the risk of bleeding while exposing the placenta to pro-thrombotic risks. Local inhibitory mechanisms-TFPI-1 and TFPI-2, thrombomodulin, annexin V and the fibrinolytic system-limit coagulation activation and fibrin deposition. Pregnancy complications have been associated with abnormalities in the functions of these inhibitors. Haemostatic processes in placental cells change throughout gestation and are affected by the changing requirements of the organ. PMID- 12763487 TI - Risk factors for thrombosis in pregnancy. AB - Pregnancy is a hypercoagulable state affecting both the coagulation and the fibrinolytic systems. Any exacerbation of the pre-disposing factors for coagulation may well lead to a thrombotic event more often in pregnant women than in the general population. Arterial thrombosis is very rare in pregnancy. Pre eclampsia may be a risk factor for the development of arterial disease in later life. Venous thromboembolism (VTE) in pregnancy, although still rare, is a major cause of maternal mortality. Risk factors, such as older age, increased weight and emergency Caesarean section, as well as acquired and genetic thrombophilia, often coexist and reinforce each other. Appropriate thromboprophylaxis needs to be considered and applied on an individual basis. Uteroplacental thrombosis provides a common pathophysiological link between various poor pregnancy outcomes, including recurrent miscarriage, stillbirth, placental abruption, fetal growth restriction and pre-eclampsia. Its significance depends on the gestational age. Acquired and genetic thrombophilia may be associated with such conditions, particularly in early-onset disease. More data are required to assess the significance of such thrombophilias in obstetric practice. Any treatment should be in the context of clinical trials. PMID- 12763488 TI - Antiphospholipid antibodies and pregnancy. AB - Antiphospholipid antibodies, i.e. lupus anticoagulants and anticardiolipin antibodies, are associated with obstetric complications. Fetal death and recurrent spontaneous abortions represent the obstetric criteria of the antiphospholipid syndrome. They occur with similar frequences and have an overall prevalence of 15-20%. Lupus anticoagulants carry a risk 3.0 to 4.8 times, and anticardiolipin antibodies 0.86 to 20 times higher than controls. The mechanism(s) by which antiphospholipid antibodies cause these events still has to be defined: thrombosis in the placental vessels, and impairment of embryonic implantation have been proposed. Unfractionated or low-molecular-weight heparin, alone or in combination with low-dose aspirin, represent the current standard treatment of pregnant antiphospholipid-positive women for preventing recurrent obstetric complications. Upon treatment, the live birth rate increases from 0-40% to 70-80%. However, there is still an excessive frequency of maternal and/or fetal complications, indicating the necessity of a better calibration of the dosage, duration and timing of administration of heparin treatment. PMID- 12763489 TI - Thrombocythaemia and pregnancy. AB - The diagnosis of essential thrombocythaemia (ET) is considered when all causes of reactive (secondary) thrombocytosis have been excluded in a patient with a persistent thrombocytosis. Among the chronic myeloproliferative disorders, ET has the highest proportion of young as well as female patients affected. As such, the optimal management of ET during pregnancy becomes an important issue. Although ET is not a contraindication to childbearing, the risks to both the fetus and mother are increased, and would-be-parents should be counselled regarding these. Obstetric complications occurring at higher than expected rates include spontaneous abortion in the first-trimester. Maternal complications, both haemorrhagic and thrombotic, are reported relatively infrequently. It is generally difficult to make specific management recommendations because of the relative rarity of the association between ET and pregnancy, which has precluded adequately powered controlled trials. In this communication, we review our own experience as well as that of other investigators on the subject matter and provide management guidelines that are based on best available information. PMID- 12763490 TI - Inherited thrombophilia and gestational venous thromboembolism. AB - Thromboembolic disease is a leading cause of maternal morbidity and mortality during pregnancy and the puerperium. To reduce the incidence of venous thromboembolism in pregnancy and improve outcomes, an individual risk stratification based on probability of thrombosis as a rationale for an individual risk-adapted prophylaxis is required. Within the past 10 years, a significant improvement in risk estimation has been achieved due to the identification of new genetic risk factors of thrombosis. In women without prior thrombosis, the presence of a heterozygous factor V Leiden or heterozygous G20210A mutation in the prothrombin gene is associated with a pregnancy associated thrombotic risk of approximately 1 in 400. Thus, in pregnant carriers of either one of these mutations the risk of venous thromboembolism is low- indicating that pregnancy-associated thrombosis is multicausal, resulting from the interaction of combined defects. A combination of the two genetic risk factors can increase the risk to a modest level (risk 1 in 25). In women with a single episode of prior thrombosis associated with a transient risk factor, for example, surgery or trauma, and no additional genetic risk factor, the probability of a pregnancy-associated thrombosis appears also to be low. In contrast, in women with a prior idiopathic venous thrombosis who carry an additional hereditary risk factor or who have a positive family history of thrombosis, a high risk (>10%) can be expected, supporting the indication for active antepartum and postpartum heparin prophylaxis. Despite the remarkable progress in risk stratification, the absolute magnitude of risk in many cases is unknown and current recommendations remain empirical. PMID- 12763491 TI - Prevention of venous thromboembolism in pregnancy. AB - Pulmonary thromboembolism, rising from deep venous thrombosis (DVT), is a major cause of maternal death in the developed World. DVT is a significant source of morbidity in pregnancy and the puerperium with long-term sequelae such as post thrombotic syndrome. The major risk factors for venous thromboembolism (VTE) are: increasing age, particularly over 35 years; operative vaginal delivery; Caesarean section, especially emergency Caesarean section in labour; high body mass index; previous VTE, especially if idiopathic or thrombophilia-associated; thrombophilia; and a family history of thrombosis suggestive of an underlying thrombophilia. Thromboprophylaxis centres largely on the use of low-molecular weight heparin (LMWH). LMWHs, such as enoxaparin and dalteparin, have substantial clinical and practical advantages compared with unfractionated heparin, particularly in terms of improved safety with a significantly lower incidence of heparin-induced osteoporosis and thrombocytopenia. Such agents should be used in women with significant risk factors for VTE both antenatally and post-partum. PMID- 12763492 TI - Diagnosis and treatment of venous thromboembolism in pregnancy. AB - Suspected or confirmed venous thromboembolism (VTE) (deep-vein thrombosis and pulmonary embolism) in non-pregnant patients are common clinical problems with ample clinical research upon which diagnostic and treatment recommendations are based. Unfortunately, the level of complexity is increased in the diagnostic and therapeutic management of pregnancy-associated VTE by evolving physiological changes in expectant mothers, the effects of diagnostic and therapeutic management on the unborn child and the lack of validation of these management strategies in pregnancy. This chapter considers the epidemiology, pathogenesis, diagnosis and treatment of pregnancy-associated VTE. It highlights the poverty of research upon which to base clinical recommendations for this common problem yet offers practical but conservative approaches to patients with suspected and confirmed pregnancy-associated VTE. PMID- 12763493 TI - Arterial thromboembolism in pregnancy. AB - Arterial occlusion resulting from primary thrombus formation in an artery or due to embolization from a site elsewhere in the circulation is uncommon in women of childbearing age. Myocardial infarction, stroke and peripheral arterial occlusion are rare in pregnant or puerperal women. Although atherosclerosis is the most common cause of arterial thromboembolism in the general population, other mechanisms--for example, prosthetic heart valves and drugs which cause vasospasm- are also important in young and pregnant patients. The clinical sequelae of arterial thromboembolism include sudden death and significant long-term morbidity. The best management must be the recognition of women at risk and, where possible, risk reduction and the introduction of measures to prevent acute events. PMID- 12763494 TI - Inherited thrombophilia and pregnancy loss. AB - A growing body of evidence obtained during the past 6 years suggests a significant role for inherited thrombophilia in the development of gestational vascular complications. Case-control and cross-sectional studies have demonstrated that thrombophilia is more prevalent in cohorts of women with pregnancy loss. Placental pathological findings in women with thrombophilia are hallmarked by thrombosis and fibrin deposition. Preliminary case-control studies suggest that low-molecular-weight heparins (LMWH) are effective in preventing pregnancy loss in women with thrombophilia and previous fetal wastage. PMID- 12763495 TI - Inherited thrombophilia and gestational vascular complications. AB - The most common causes of inherited thrombophilia, the factor V Leiden and the factor II A20210 mutations, confer a higher risk of venous thromboembolism. Moreover, several studies have suggested that they can have a role in the occurrence of gestational vascular complications in otherwise unexplained recurrent fetal losses, hypertensive disorders of pregnancy and fetal growth restriction. Observational and case-control studies addressing these issues are available in literature. However, longitudinal, perspective studies are lacking. Mild hyperhomocysteinaemia can be due partly to inherited susceptibility--as the homozygous carriership of the T677 variant in the gene encoding 5,10 methylenetetrahydrofolate reductase (MTHFR). Case-control studies have been carried out on a possible association between unexplained fetal losses and mild hyperhomocysteinaemia. Although case-control and perspective studies are available on hyperhomocysteinaemia and other gestational vascular complications the data are conflicting.Intervention studies have been carried out to prevent adverse obstetric outcomes in women with factor V Leiden or factor II A20210 mutations and previous adverse outcomes. Although these are not randomized controlled trials, all have found significantly better outcomes in treated pregnancies compared to those of untreated pregnancies in the same women. PMID- 12763496 TI - Maternal thrombophilia and neonatal thrombosis. AB - In neonates and infants, numerous clinical and environmental conditions lead to elevated thrombin generation and subsequent thrombus formation. Genetic prothrombotic defects (protein C, protein S and antithrombin deficiency, mutations of coagulation factor V and factor II, elevated lipoprotein (a)) have been established as risk factors of thromboembolic events in neonates and infants. The interpretation of the laboratory evaluation relies on age-dependent normal reference values. Because appropriate clinical trials are missing in these age groups, treatment recommendations are adapted from small-scale studies in neonates and infants and from guidelines relating to adult patient protocols. Secondary long-term anticoagulation should be administered on an individual basis. PMID- 12763499 TI - Stress-related mucosal disease in critically ill patients. AB - Stress-related mucosal disease is common in critically ill patients and can result in significant morbidity. The pathophysiology of this condition is multifactorial but mucosal ischaemia and gastric acid have pivotal roles. The major risk factors for developing stress-related mucosal bleeding are prolonged mechanical ventilation and coagulopathy. The mainstay of clinical management is prevention with acid-suppressing medications and cytoprotective agents. This chapter discusses medications used to prevent this condition and suggests an approach for management. Recent developments in gastroenterology include the development and use of proton pump inhibitors and the discovery of the association Helicobacter pylori with gastritis. The role of each of these in stress-related mucosal disease is discussed. Finally, an approach to the bleeding critically ill patient is presented. PMID- 12763500 TI - Gastrointestinal disorders of the critically ill. Shock and acute pancreatitis. AB - Shock and pancreatitis are closely associated. Shock can be a sequel of severe pancreatitis and systemic shock may induce pancreatitis. This chapter discusses both features with regard to their clinical and pathophysiological characteristics. PMID- 12763501 TI - Gastrointestinal disorders of the critically ill. Cholestasis of sepsis. AB - Cholestasis of sepsis is a form of hepatocellular cholestasis that occurs as a result of sepsis. Usually, prior to the development of cholestasis, the manifestations of sepsis dominate the clinical picture. The occurrence of cholestasis is without direct bacterial involvement of the biliary system and appears to be mediated systemically by pro-inflammatory cytokines. These cytokines are released in response to the vigorous inflammatory reaction mediated by endotoxinaemia and bacterial wall lipopolysaccharides. The principal cytokines involved are the pro-inflammatory tumour necrosis factor-alpha (TNF-alpha), interleukin (IL) 1-beta and IL-6. Interplay between these cytokines and a series of hepatocyte membrane transporters appears to result in the cholestasis. Management principles focus upon the control of sepsis. PMID- 12763502 TI - Gastrointestinal disorders of the critically ill. Shock liver. AB - Shock liver describes a collecting pool of critically ill patients in whom the elevation of liver function tests or overt hepatic dysfunction is apparent. Different grades of shock liver affect about 50% of all intensive-care patients, varying from a mild elevation of serum aminotransferase and bilirubin levels in septic patients to an acute onset of high serum aminotransferases after haemodynamic shock. Abnormalities can subside within days or progressively deteriorate when persistent hepatic microcirculatory failure is present. Although hepatic injury in critically ill patients influences mortality rates it is underdiagnosed. The underlying pathophysiology involves changes in the portal and arterial blood supply as well as in microcirculation. Cross-talk between hepatocytes, Kupffer cells and endothelial cells, leading to an inflammatory response mediated primarily by tumour necrosis factor-alpha (TNF-alpha), is central to shock liver. The liver is a victim of shock inducers, and can also be the orchestrator of the inflammatory response syndrome (IRS). Hepatic injury by TNF-alpha is modulated by the prevalent pro-inflammatory or anti-inflammatory mediator profile elaborated by Kupffer cells. Kupffer cells additionally participate in the clearance of endotoxin, bacteria and inflammatory mediators and are thereby capable of preventing IRS. The hepatocyte undergoes dramatic alterations in synthetic activity, biliary transport, bile flow and glucose metabolism. Although standard determinations of aminotransferases, coagulation studies, glucose, lactate and bilirubin can detect hepatic injury they only partially reflect the cellular mechanisms driving shock liver. The management of shock liver is focused on the prevention of precipitating causes by controlling sepsis, circulation parameters and metabolism in addition to the cautious monitoring of therapeutic measures that can increase hepatic injury, which include intravenous nutrition, mechanical ventilation and catecholamine administration. PMID- 12763503 TI - Gastrointestinal disorders of the critically ill. Biliary sludge and cholecystitis. AB - Biliary sludge develops commonly in critically ill patients and may be associated with biliary colic, acute pancreatitis or acute cholecystitis. Sludge often resolves upon resolution of the underlying pathogenetic factor. It is generally diagnosed on sonography. Treatment of sludge itself is unnecessary unless further complications develop. Acute acalculous cholecystitis also develops frequently in critically ill patients. It may be difficult to diagnose in these patients, manifesting only as unexplained fever, leukocytosis or sepsis. Sonography and hepatobiliary scintigraphy are the most useful diagnostic tests. Management decisions should take into account the underlying co-morbid conditions. For many patients, percutaneous cholecystostomy may be the best management option. Cholecystostomy may also provide definitive drainage as patients recover and underlying critical illness resolves. PMID- 12763504 TI - Gastrointestinal disorders of the critically ill. Bacterial translocation in the gut. AB - The human gastrointestinal tract is colonized by a dense population of microorganisms, referred to as the bacterial flora. Although the gut provides a functional barrier between these organisms and the host, bacterial translocation is a common event in the healthy person. However, in critically ill patients, with various underlying diseases, this bacterial translocation may lead to infections and consequently to a further reduction in general health status. The mechanism of bacterial translocation is widely, and somehow controversially investigated in vitro and in animal models. In human studies, several diseases have been associated with bacterial translocation. However, methodological shortcomings, insufficient populations and conflicting results leave many open questions. This is also reflected in the various published therapeutic strategies. To overcome this problem more investigations in humans are needed, especially in techniques for detecting bacterial translocation. PMID- 12763505 TI - Pseudo-obstruction in the critically ill. AB - Intestinal pseudo-obstruction is defined as a clinical syndrome characterized by impairment of intestinal propulsion, which may resemble intestinal obstruction, in the absence of a mechanical cause. It may involve the small and/or the large bowel, and may present in acute, subacute or chronic forms. We have performed a systematic review of acute pseudo-obstruction, also referred to as Ogilvie's syndrome in the literature, and focused on proposed mechanisms, manifestations and management of post-surgery and critically ill patients who suffer from one or more underlying clinical conditions. The hallmark of the syndrome is massive intestinal distension, which is detected on clinical inspection and plain abdominal radiography. The underlying pathophysiological mechanisms are not fully understood. Therefore, treatment focuses on preventing intestinal perforation, which is associated with an average 21% mortality rate. PMID- 12763506 TI - Gastrointestinal disorders of the critically ill. Systemic consequences of ileus. AB - Ileus refers to the partial or complete blockage of the small and/or large intestine either by functional (adynamic or paralytic ileus) or mechanical bowel obstruction. The diffuse gastrointestinal dysmotility during functional and mechanical ileus may result in intestinal dilatation, increased luminal pressure and gut wall ischaemia which may lead to increased intra-abdominal pressure (IAP). Any type of ileus may promote abdominal fluid sequestration with severe systemic hypovolaemia, intestinal bacterial overgrowth with the evolution of bacterial translocation and systemic invasive infections and inflammation of the intestinal wall with concomitant release of cytokines and the development of the systemic inflammatory response syndrome. The most serious complications of ileus are mediated by an increase in IAP. Intra-abdominal hypertension has been found in up to 20% of critically ill patients and may lead to a broad pattern of systemic consequences with multiple organ dysfunction, including cardiovascular, hepatic, pulmonary, renal and neurological function. The abdominal compartment syndrome is an emergency condition which is defined as elevation of IAP above 20 to 25 mmHg and the presence of systemic consequences. Therapeutic considerations include the maintenance of adequate hydration status, avoidance of drugs known to impair intestinal perfusion, stimulation of gastric and intestinal motility and various nutritional aspects. Colonic tube placement after decompressive colonoscopy may be effective in reducing intestinal dilatation. In the abdominal compartment syndrome the 'open abdominal approach' with decompressive laparotomy by opening the peritoneal cavity and temporary abdominal closure is the therapy of choice. PMID- 12763507 TI - Non-occlusive mesenteric ischaemia: a common disorder in gastroenterology and intensive care. AB - Non-occlusive mesenteric ischaemia is characterized by gastrointestinal ischaemia with normal vessels. In gastroenterology it is recognized as rare disease occasionally causing acute bowel infarction or ischaemic colitis. From intensive care literature this disorder is recognized as an early phenomenon during circulatory stress. This early mucosal ischaemia then leads to increased permeability, bacterial translocation, and further mucosal hypoperfusion. The damage is produced mainly during reperfusion following ischaemia with fresh inflow of oxygen and outflow of waste products into the systemic circulation. The mechanisms underlying non-occlusive mesenteric ischaemia include macrovascular vasoconstriction, hypoperfusion of the tips of the villi and shunting. It is very common in critically ill and perioperative patients, but also occurs in pancreatitis, renal failure and sepsis. Treatment options include aggressive fluid resuscitation and careful choice of vasoactive drugs. Control of reperfusion damage and new endothelin-antagonists are potentially useful new treatment options. PMID- 12763508 TI - Gastrointestinal disorders and the critically ill. Clostridium difficile infection and pseudomembranous colitis. AB - Clostridium difficile causes a spectrum of diseases ranging from diarrhoea to pseudomembranous colitis, primarily in the hospitalized elderly, although community-acquired infection is probably under-documented. Host factors are increasingly recognized as critical determinants of disease expression. Exposure to antibiotics, particularly those adversely affecting anaerobic gut flora, appears to create a niche which is exploited by C. difficile. Several retrospective and intervention studies have indicated that third-generation cephalosporins have a high propensity to induce C. difficile diarrhoea. Conversely, some broad-spectrum antibiotics, including ureidopenicillins (e.g. piperacillin-tazobactam) and ciprofloxacin, are less likely to induce C. difficile infection. Effective control of C. difficile in the hospital requires both antibiotic control and prevention of environmental seeding and bacterial spread. Epidemic C. difficile strains are widely distributed in the hospital environment, both as a cause and result of nosocomial diarrhoea. Current treatment options are antibiotic-based, which is less than ideal. Although many biotherapeutic approaches have been tried few have shown real benefit. PMID- 12763509 TI - Medical choices available for management of menopause. AB - The indications for hormone therapy (HT) have changed markedly since the 1980s; they now include the treatment of menopausal symptoms and the prevention and treatment of osteoporosis in the short term. Long-term therapy is discouraged because of the small increase in risk of breast cancer after 5 years of therapy. Careful assessment of the midlife woman allows for individualized risk-benefit analysis with the formulation of a specific health management plan. Lifestyle advice and modification form the cornerstone of management-followed by therapeutic options if appropriate indications exist. In some industrialized countries alternative therapies are preferred despite little scientific evidence of their efficacy. The choices of hormonal products have increased, with the introduction of new formulations and routes of administration allowing for more optimal treatment of the menopause, especially in the presence of concurrent medical conditions, for example, diabetes, breast cancer or fibroids. PMID- 12763510 TI - Psychological aspects of menopause management. AB - The influence of endocrine function on the mood of the menopausal woman continues to be debated and researched. While many women present at the menopause with depression and anxiety, the reasons for these mood disorders cannot be attributed to menopause status alone. The influence of psychological factors, lifestyle, body image, interpersonal relationships, role, and sociocultural factors in predicting levels of depression and anxiety in the menopausal patient cannot be ignored. This chapter discusses the research to date on menopause and depression and anxiety. Included is a discussion of the role of psychosocial factors in the symptomatology of perimenopausal, post-menopausal and prematurely menopausal women. The importance of understanding the individual menopausal experiences of women within the context of their lives while offering support, education, and validation is highlighted. The need for a multidimensional approach to treating the menopausal woman who presents with mood disorders is examined. Finally, this chapter makes it clear that psychosocial aspects of menopause management require further research, particularly on the experiences of women who enter menopause prematurely. PMID- 12763511 TI - The menopausal transition: characteristics and management. AB - The menopausal transition is a complex period in a woman's life, reflecting ovarian ageing and concomitant hormonal changes, in addition to social and metabolic changes. These changes, in turn, influence the signs and symptoms common to this period. Symptoms which are influenced by the hormonal fluctuations occurring during the menopausal transition include vasomotor symptoms and vaginal dryness; others are breast tenderness, poor sleep and pre-menstrual dysphoria. Hormonal therapy has been shown to be first-line therapy for many of these symptoms. Other types of pharmacotherapies may be helpful, including selective serotonergic uptake inhibitors for vasomotor symptoms. Characteristic signs of the menopausal transition include abnormal uterine bleeding, best managed with hormonal pharmacotherapy; diminishing bone mineral density, which may warrant diagnostic intervention, and may benefit from dietary and lifestyle modifications; and increased body-mass index and worsening lipid profile, which also may benefit from dietary and lifestyle modifications. PMID- 12763512 TI - The menopause and HRT. Prevention and management of osteoporosis. AB - Post-menopausal osteoporosis is characterized by increased fracture risk due to deficiencies in both the quantity and quality of bone. Assessing fracture risk involves combining clinical risk factors, including fall risks, with bone density testing. Treatment strategies are aimed at reducing fracture risk. General nutritional and lifestyle measures are appropriate for all women. Drug treatment is most clearly indicated in post-menopausal women at high current fracture risk. Treatment should also be considered for women at intermediate fracture risk, including those who have both low bone density and other risk factors for fracture. Whether there is practical clinical value in treating low-risk patients is much less clear. Non-pharmacological approaches addressing the consequences of fractures are integral parts of a comprehensive treatment programme. Reducing both the frequency and the effects of falls complements the efforts of treating osteoporosis to reduce the incidence of fractures and their important clinical consequences. PMID- 12763513 TI - The menopause and HRT. Hormone replacement therapy, cardiovascular and cerebrovascular disease. AB - Historically, research into vascular effects of hormone replacement therapy (HRT) poignantly highlights the difficulties in extrapolating from medical research into clinical practice. Original observational trials encouraged great enthusiasm that HRT was protective against coronary heart disease (CHD) in post-menopausal women. This was supported by a plethora of beneficial mechanistic effects of HRT on the vascular system. In contrast, recent controlled trials have shown that a specific combined oral HRT actually causes a small increased risk of cardiovascular and cerebrovascular events, compared to placebo. The absolute risks were small, yet were accompanied by increased venous thrombosis and breast cancer. Although many controversies still remain regarding the vascular effects of specific types of HRT, currently, in my opinion, all HRT should be considered as causing a small increased risk of vascular events, until proven otherwise. However, the apparent lack of HRT effectiveness in the prevention of vascular disease should not deter enthusiasm for the use of established preventive therapies, especially lifestyle measures. PMID- 12763514 TI - The menopause and HRT. Urogenital effects of hormone therapy. AB - The urogenital tract and lower urinary tract are sensitive to the effects of oestrogen and progesterone throughout adult life. Epidemiological studies have implicated oestrogen deficiency in the aetiology of lower urinary tract symptoms occurring following the menopause. Although to date the role of oestrogen replacement therapy in the management of post-menopausal urinary incontinence remains controversial, its use in the management of women complaining of urogenital atrophy is now well established. This aim of this chapter is to review the recent evidence regarding the urogenital effects of hormone therapy with a particular emphasis on the management of post-menopausal urinary incontinence, recurrent lower urinary tract infections and urogenital atrophy. PMID- 12763515 TI - The menopause and HRT. HRT and cognitive decline. AB - It is biologically plausible that hormone replacement therapy (HRT) would be protective against cognitive decline and Alzheimer's disease (AD). We review observational and randomized trials to determine whether HRT might protect against cognitive decline in cognitively unimpaired and demented women. We also address issues of clinical relevance, including duration and type of treatment and patient characteristics, including type of menopause (surgical versus natural), age, education and menopausal symptoms. Differences in participant characteristics and testing methods limit the ability to draw conclusions across randomized studies of HRT in non-demented women. The available evidence suggests no detrimental effect of HRT on cognitive function and inconsistent benefits on verbal memory and reasoning, frontal functions and speeded attention. Meta analyses of observational trials suggest that HRT protects against the development of AD, but randomized trials indicate no long-lasting benefit in patients with AD. Evidence is insufficient to recommend HRT to maintain cognitive function. PMID- 12763516 TI - Breast cancer and post-menopausal hormone therapy. AB - From the introduction of post-menopausal hormone replacement therapy (HRT) there has been great concern that HRT could possibly increase the risk of breast cancer. Prolonged exposure to endogenous oestrogens undeniably increases the risk of breast cancer. Questions that are important and until now only partly answered, are the following. Are oestrogens tumour promoters, as they induce mitosis, lead to proliferation and, therefore, accelerated growth of clinically occult pre-existing tumours? In addition to this, are they genotoxic mutagenic carcinogens, or could they initiate tumours by way of accumulation of incessant DNA-replication damage mechanism? Opinions vary as to the effect of the addition of a progestogen. There is a multitude of different progestogens which could bind with differing affinity to progesterone receptor PR-A or PR-B, and which have different physiological functions via differential gene regulation. The action of a progestogen on the oestrogen-induced cellular mitotic activity could be synergistic or antagonistic (by different pathways: oestrogen receptor downregulation, activating of metabolic pathways within the breast or stimulation of apoptosis)? Over 60 observational studies and two randomized trials provide evidence that the small but significant increase in risk appears with long-term current post-menopausal hormone use. The addition of a progestogen does not decrease the risk as seen with oestrogens alone and might increase the risk further. It is not clear whether there is a difference in risk with sequentially combined versus continuously combined HRT. Many questions nevertheless still remain. Is the risk increase limited to lean women only? What about risk modifying factors such as alcohol use and a positive family history for breast cancer? Are tumours detected under HRT less aggressive, is there a better prognosis and is the mortality not increased while morbidity is? And is HRT contraindicated for women with a positive family history for breast cancer or in those women who have been treated for breast cancer? And finally, are there alternative options for these women? PMID- 12763517 TI - Hormone replacement therapy and endometrial, ovarian and colorectal cancer. AB - Sex-steroid-related tumours in women are represented by breast cancer and endometrial cancer, but a possible relationship may exist between sex steroids and both ovarian and colon cancer. Unopposed oestrogen therapy is known to increase the risk of endometrial cancer and is appropriate only for hysterectomized women. In women with an intact uterus, an appropriate combination of oestrogen and progestin does not appear to increase-and may even decrease-the risk of endometrial cancer. Current users of HRT seem to benefit from a reduced risk for colon cancer. As for epithelial ovarian cancer, the present data are very conflicting. The association between replacement hormones and this malignancy seems to be stronger for unopposed oestrogen than for oestrogen progestin treatment. Data available at the moment do not allow discriminating for dose, routes of administration, bioavailability and tissue effects of different compounds so that it is inappropriate to consider all forms of HRT jointly. The future of HRT in post-menopausal women lies in the individualization of the therapy based upon personal risk factors and characteristics. PMID- 12763518 TI - Hormone therapy and venous thromboembolism. AB - Convincing data from randomized trials and observational studies have demonstrated a two- to threefold increased risk of venous thromboembolism (VTE) with the use of hormone replacement therapy (HRT) in post-menopausal women. This risk is highest in the first year of use, but an increased risk persists after the first year if HRT use is ongoing. The risk of VTE is increased for oral oestrogen alone, oral oestrogen combined with progestin and probably for transdermal HRT. There is an increase in both idiopathic and non-idiopathic VTE with HRT. Early evidence suggests an interaction of HRT with thrombophilic states such as the factor V Leiden mutation, resulting in a synergistic increase in the risk of VTE. There is also an increased risk of VTE with raloxifene and tamoxifen, but the effects of low-dose HRT and tibolone on VTE risk are less clear. PMID- 12763519 TI - The role of androgen therapy. AB - The concept of a female androgen insufficiency syndrome, although not new, remains somewhat controversial. Androgens are quantitatively the predominant sex steroid in women, circulating in the micromolar and nanomolar concentration range, compared with picomolar levels of oestrogens. The most significant biologically active androgen is testosterone (T), which circulates bound tightly to sex-hormone-binding globulin (SHBG) and loosely to albumin. It is generally held that the non-SHBG-bound fraction is the bioavailable moiety. Hence, clinically useful T measurements require data on total concentrations as well as SHBG level. Testosterone insufficiency occurs in a number of circumstances, including hypopituitarism, premature ovarian failure, adrenal failure, exogenous corticosteroid use and oral oestrogen therapy (causing elevation of SHBG and suppression of gonadotrophins). Clinical symptoms of androgen insufficiency include loss of libido, diminished well-being, fatigue and blunted motivation and have been reported to respond well to T replacement, generally without significant side-effects. PMID- 12763521 TI - Emotions are building up in the field of extracellular proteolysis. AB - Activity-dependent remodeling of neural connections might require localized extracellular proteolysis. The tissue-type plasminogen activator (tPA)-plasmin proteolytic system is expressed in different regions of the central nervous system, in the context of a variety of physiological and pathological processes. Accumulating evidence regarding the expression and role of tPA and its inhibitors suggests that extracellular proteolysis is a key player in the biology of memory, emotions and neurodegeneration. PMID- 12763522 TI - CD4+ cell memory: the enigma of Th1 cells. AB - Despite a wealth of information pertaining to functional and phenotypic attributes of memory CD4(+) cells, the mechanisms that underlie the generation and persistence of memory in this subset are largely unknown. Recent work suggests that the development of memory might be differently regulated in T helper-1 and T-helper-2 cells, owing to differences in their susceptibility to cell death. These studies support a new paradigm, in which memory T cells are heterogeneous in terms of their stage of maturation and function as well as mechanisms of homeostatic control. PMID- 12763524 TI - Mitochondria in cell death: novel targets for neuroprotection and cardioprotection. AB - Post-mitotic neurons and heart muscle cells undergo apoptotic cell death in a variety of acute and chronic degenerative diseases. The intrinsic pathway of apoptosis involves the permeabilization of mitochondrial membranes, which leads to the release of protease and nuclease activators, and to bioenergetic failure. Mitochondrial permeabilization is induced by a variety of pathologically relevant second messengers, including reactive oxygen species, calcium, stress kinases and pro-apoptotic members of the Bcl-2 family. Several pharmacological agents act on mitochondria to prevent the permeabilization of their membranes, thereby inhibiting apoptosis. Such agents include inhibitors of the permeability transition pore complex (in particular ligands of cyclophilin D), openers of mitochondrial ATP-sensitive or Ca(2+)-activated K(+) channels, and proteins from the Bcl-2 family engineered to cross the plasma membrane. In addition, manipulations that modulate the expression or activity of mitochondrial uncoupling proteins can prevent the death of post-mitotic cells. Such agents hold promise for use in clinical neuroprotection and cardioprotection. PMID- 12763523 TI - Quantitative real-time PCR: a powerful ally in cancer research. AB - In this era of the Human Genome Project, quantitation of gene expression in tumor or host cells is of paramount importance for investigating the gene patterns responsible for cancer development, progression and response or resistance to treatment. Quantitative real-time PCR (qrt-PCR) technology has recently reached a level of sensitivity, accuracy and practical ease that supports its use as a routine bioinstrumentation for gene level measurement. Several applications have already been implemented in the field of cancer research, and others are being validated, showing that this molecular biology tool can provide both researchers and clinicians with precious information concerning the behavior of tumors. Knowledge of the biochemical principles underlying this biotechnology can be of great value to interpret correctly qrt-PCR data. PMID- 12763525 TI - p66(Shc): at the crossroad of oxidative stress and the genetics of aging. AB - The biology of aging has been mysterious for centuries. Removal of the p66(Shc) gene, which encodes an adaptor protein for cell signaling, extends lifespan by approximately 30% in mice and confers resistance to oxidative stress. The absence of p66(Shc) correlates with reduced levels of apoptosis. Oxidants induce phosphorylation of serine36 on p66(Shc), contributing to inactivation of members of the Forkhead transcription factor family, some of which appear to regulate the expression of antioxidant genes. The expression of p66(Shc) is regulated by the methylation status of its promoter. This leads us to hypothesize that increased methylation of the p66(Shc) promoter might contribute to the absence of its expression and therefore extended longevity in particular individuals. PMID- 12763526 TI - VIP- and PACAP-mediated immunomodulation as prospective therapeutic tools. AB - The immune system and the brain continuously signal to each other, often along the same pathways, which might explain the connection between immunity, the brain and disease. Neuropeptides and their receptors represent part of this communication network, and recent work has examined their relevance to health, proving a potentially crucial clinical significance. The structurally related neuropeptides, vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase-activating polypeptide (PACAP), are emerging as a means of fine tuning in the maintenance a balanced steady state in the immune system. Murine knockout and transgenic models for a VIP receptor suggest that VIP is an endogenous anti inflammatory mediator with characteristics resembling those of a T-helper-2 cytokine. Thus, through molecular mechanisms that are being discovered, VIP might extend the range of therapeutic treatments available for various disorders, including acute and chronic inflammatory diseases, septic shock and autoimmune diseases. PMID- 12763527 TI - Mouse models for the study of Crohn's disease. AB - Crohn's Disease (CD) is a chronic inflammatory bowel disease (IBD) that can affect any portion of the gastrointestinal tract and can cause significant morbidity. A variety of animal models of both acute and chronic intestinal inflammation have been developed to investigate disease pathogenesis and novel treatment modalities. These include chemically induced, genetically manipulated and immune-mediated models of gut inflammation, each of which possesses similarities to human IBD and offers unique advantages for studying specific aspects of disease pathogenesis. However, the majority of these models are characterized by colitis and, unlike human CD, do not involve the small intestine. More recently, murine models of chronic ileal inflammation have been characterized that spontaneously develop and closely resemble human CD with regard to disease location, histologic features and clinical response to therapy. Two mouse models of experimental ileitis will be discussed in this review: the TNF DeltaARE and SAMP1/YitFc strains. Studies using these new models might provide important insight into the pathogenesis of human CD and test the efficacy of potential therapies to treat this devastating disease. PMID- 12763528 TI - Pathogenesis of endometriosis: natural immunity dysfunction or autoimmune disease? AB - Endometriosis is a chronic inflammatory disease, characterized by implantation and growth of endometrial tissue outside the uterine cavity. This disabling condition is considered one of the most frequent diseases in gynecology, affecting 15-20% of women in their reproductive life. Pelvic endometriosis, the most common form of the disease, is associated with increased secretion of pro inflammatory cytokines, neo-angiogenesis, intrinsic anomalies of the refluxed endometrium and impaired function of cell-mediated natural immunity. Recently, endometriosis has also been considered to be an autoimmune disease, owing to the presence of autoantibodies, the association with other autoimmune diseases and recurrent immune-mediated abortion. These findings are in apparent contradiction with the reduced cell-mediated natural immunity observed during the disease. In this review, we focus on the multiple processes underlying the complex pathogenesis of endometriosis, with particular emphasis on the role played by the immune system with the induction of autoimmunity. PMID- 12763529 TI - Calculation of electrophoretic mobility in ternary solvent electrolyte systems. AB - Electrophoretic mobility of salmeterol and phenylpropanolamine in capillary electrophoresis has been determined using acetate buffer containing different concentrations of water, methanol and acetonitrile. Maximum electrophoretic mobilities for salmeterol and phenylpropanolamine have been observed at water methanol-acetonitrile (5:50:45, v/v) and (3:60:37, v/v), respectively, while minimum mobilities of both compounds occurred at methanol-acetonitrile (30:70, v/v). The generated experimental data have been used to evaluate a mathematical model to compute the electrophoretic mobility of the analytes. The proposed model reproduced the mobility data with mean percentage deviations within 1-4%. PMID- 12763530 TI - Determination of ambroxol in human plasma using LC-MS/MS. AB - A sensitive and selective liquid chromatographic method coupled with tandem mass spectrometry (LC-MS/MS) was developed for the quantification of ambroxol in human plasma. Domperidone was used as internal standard, with plasma samples extracted using diethyl ether under basic condition. A centrifuged upper layer was then evaporated and reconstituted with 200 microl methanol. The reconstituted samples were injected into a C(18) XTerra MS column (2.1 x 30 mm) with 3.5 microm particle size. The analytical column lasted for at least 600 injections. The mobile phase was composed of 20 mM ammonium acetate in 90% acetonitrile (pH 8.8), with flow rate at 250 microl/min. The mass spectrometer was operated in positive ion mode using turbo electrospray ionization. Nitrogen was used as the nebulizer, curtain, collision, and auxiliary gases. Using MS/MS with multiple reaction monitoring (MRM) mode, ambroxol was detected without severe interferences from plasma matrix. Ambroxol produced a protonated precursor ion ([M+H](+)) at m/z 379 and a corresponding product ion at m/z 264. And internal standard (domperidone) produced a protonated precursor ion ([M+H](+)) at m/z 426 and a corresponding product ion at m/z 174. Detection of ambroxol in human plasma was accurate and precise, with quantification limit at 0.2 ng/ml. This method has been successfully applied to a study of ambroxol in human specimens. PMID- 12763531 TI - A biochromatographic framework to evaluate the calcium effect on the antihypertensive molecule-human serum albumin binding. AB - The Ca(2+) cation effect on the antihypertensive molecule binding on human serum albumin (HSA) was studied by biochromatography. The thermodynamic parameters corresponding to this binding were determined for a wide range of Ca(2+) concentration (x). For the two antihypertensive molecules under study, their binding to HSA can be divided into two Ca(2+) cation concentration regions due to a HSA phase transition. This result was confirmed by an enthalpy-entropy investigation. For a low x value (below x(c)=1.6 mmol l(-1)), the HSA cavity was in an ordered solid-like state leading to an increase in the interactions between the antihypertensive drugs and the HSA cavity and consequently, a solute-HSA affinity increase. For x above x(c), the HSA cavity was in a disordered solid like state, implying a decrease in the antihypertensive drug-HSA binding. PMID- 12763532 TI - Determination of allopurinol by micelle-stabilised room-temperature phosphorescence in real samples. AB - A very simple, rapid and highly sensitive method has been developed for the determination of allopurinol. The method is based on the room temperature phosphorescence of allopurinol in sodium dodecylsulphate (SDS) micelles, with thallium (I) providing the external heavy atom and sodium sulphite acting as the oxygen scavenger. Under the optimum experimental conditions, the range of application is 0.25-7.0 microg ml(-1) and the limit of detection is 0.014 microg ml(-1). The most relevant characteristic of this method is its great selectivity, e.g. allopurinol can be determined in the presence of its metabolite, oxypurinol. The results of the analysis of several pharmaceutical preparations were satisfactory. The clinical applicability of this procedure has been tested by analysing allopurinol in urine samples. PMID- 12763533 TI - Fast separations on monolithic silica columns: method transfer, robustness and column ageing for some case studies. AB - Six separation methods, developed on conventional silica high performance liquid chromatography (HPLC) columns were transferred to monolithic silica columns of 5 and 10 cm length. The transferred methods include the separation of an alkylbenzene mixture, the separations of drugs from their impurities (nimesulide, tetracycline, phenoxymethylpenicillin and erythromycin) and the separation of a green tea extract. The transfer of the first three methods was successful while for the latter three it was not. Increasing the flow rate up to 9 ml/min (where possible) inversely decreased the analysis time of the successfully transferred methods to 48 s (alkylbenzene mixture) 1.8 min (nimesulide mixture) and 3 min (tetracycline mixture) while still reasonable well separated peaks were obtained. The robustness and repeatability of the transferred and accelerated separations was found to be acceptable. Despite the use of flow rates up to 9 ml/min and frequent mobile phase changes with pH values varying from 3.5 to 7, the column performance was found to be rather constant and the column ageing to be minimal. PMID- 12763534 TI - Disposable electrochemical sensor for rapid determination of heavy metals in herbal drugs. AB - Analysis of herbal drugs and extracts need rapid and affordable methods to assure the quality of products. The application of the electrochemical sensors in the field of quality control of herbal drugs, herbal drug preparations and herbal medicinal products appears very promising, advantageous and alternative to conventional methods due to their inherent specificity, simplicity and for the fast response obtained. This paper presents a proposal about the application of disposable electrochemical sensors associated with electroanalytical instrumentation for the detection of heavy metal analysis in herbal drugs. In particular samples of St. John's wort were analysed applying anodic stripping voltammetry. The content of Cd and Pb were evaluated. The ICP spectroscopy was used as reference method. PMID- 12763535 TI - Derivative spectrophotometry in the analysis of mixtures of cefotaxime sodium and cefadroxil monohydrate. AB - Derivative spectrophotometry (ratio-spectra 1st- and 2nd-derivative and zero crossing 2nd-derivative techniques) was applied for the determination of some cephalosporins in two component mixtures. Cefotaxime sodium salt (C(16)H(16)O(7)N(5)S(2)Na) and cefadroxil monohydrate (C(16)H(17)N(3)O(5)S.H(2)O) were examined. In all procedures, the calibration plots are linear up to 43 microg/ml of each antibiotic, with r ranging from 0.9997 to 0.9999. In the ratio spectra method, the measurements were taken at 239.5 and 291.5 nm (cefotaxime, 1st-derivative), 238 and 283 nm (cefadroxil, 1st-derivative), 284 and 303 nm (cefotaxime, 2nd-derivative), and 229.5 and 245.5 nm (cefadroxil, 2nd derivative). Detection limits at P=0.05 level of significance, calculated by a statistical treatment of calibration data, ranged from 0.15 to 0.58 microg/ml. LOD and LOQ ranged, respectively, from 0.19 to 0.51 and from 0.63 to 1.70 microg/ml. By the zero-crossing 2nd-derivative method, lines of regression are linear at 257 and 279 nm (cefotaxime) and 242 and 296 nm (cefadroxil). Detection limits from 0.28 to 0.51 microg/ml. LOD and LOQ from 0.27 to 0.41 and from 0.90 to 1.37 microg/ml, respectively. All the samples were tested for stability in solution and in the course of actual analysis, up to 80 h from their preparation. The developed derivative spectrophotometric methods were applied to synthetic mixtures and the RSD values ranged between 0.05 and 1.35% (ratio-spectra technique) and 0.01 and 1.07% (zero-crossing technique). The methods were also applied to vials and tablets for these drugs. The recoveries obtained were between 100.9 and 102.4% (ratio-spectra) and between 99.8 and 102.0% (zero crossing). The procedures are simple, rapid, and did not require any preliminary separation or treatment of the samples. Instrumentation commonly available was utilised. The cephalosporins analysed are frequently used antibiotics of relevant clinical and pharmacological importance; hence this work would be of interest for the readers of journals devoted to pharmaceutical and biomedical analysis. PMID- 12763536 TI - Electrochemical study of hydroxychloroquine and its determination in plaquenil by differential pulse voltammetry. AB - Hydroxychloroquine (HCQ) is a halogenated aminoquinoline that presents wide biological activity, often being used as an antimalarial drug. The electrochemical reduction of HCQ was investigated by cyclic voltammetry and chronoamperometry using glassy carbon electrodes. By cyclic voltammetry, in acid medium, only the cathodic peak was observed. The electrochemical behavior of this peak is dependent on pH and the electrodic process occurs through an E(r)C(i) mechanism. The electron number (1e) consumed in the reduction of HCQ was obtained by chronoamperometry. A method for the electrochemical determination of HCQ in pharmaceutical tablets was developed using differential pulse voltammetry. The detection limit reached was 11.2 microg ml(-1) of HCQ with a relative standard deviation of 0.46%. A spectrophotometric study of HCQ has been also carried out utilizing a band at 343 nm. The obtained detection limit and the relative standard deviation were 0.1 microg ml(-1) and 0.36%, respectively. The electrochemical methods are sufficiently accurate and precise to be applied for HCQ determination, in laboratorial routine, which can be used to determine the drug at low level. PMID- 12763537 TI - Spectrophotometric and LC determination of two binary mixtures containing pyridoxine hydrochloride. AB - Several methods are developed for the determination of two binary mixtures containing pyridoxine HCl together with either metoclopramide HCl (mixture 1) or meclozine HCl (mixture 2). The resolution of binary mixture of pyridoxine HCl and metoclopramide HCl has been accomplished by using partial least squares (PLS-1) and principal component regression (PCR) applied to zero and first order UV spectra of the mixture, respectively. In addition HPLC method depending on using RP18 column with a mobile phase consisting of acetonitrile-water (30:70, v/v, pH 3.2) with UV detection at 305 nm was developed. In mixture 2, the simultaneous determination of pyridoxine HCl and meclozine HCl was carried out by using graphical (second derivative of the ratio spectra) and numerical spectrophotometric methods (principal component regression and partial least squares, PLS-1 and PLS-2, applied to the zero order UV spectra of the mixture). The proposed methods were successfully applied for the determination of the two binary combinations in synthetic mixtures and in commercial tablets and syrup containing several light absorbing excipients. PMID- 12763540 TI - Control of the illegal use of clenbuterol in bovine production. AB - This study is based on a plan of collecting different matrices (hair, eye, muscle, liver and kidney) in order to define a strategy for the control of the illegal use of clenbuterol in bovine production. Of all matrices utilised, hair is recommended for the analytical control of clenbuterol in living animals, due to its being permanently available and easy to collect. The eye, or rather the retina, is the matrix which gives the most trustworthy result, after the animal slaughter, and the one that best helps in the determination of the illegal use of clenbuterol in a perspective of gradual food safety improvement. PMID- 12763539 TI - On-line SPE-RP-LC for the determination of insulin derivatives in biological matrices. AB - An automated and on-line solid-phase extraction (SPE)-liquid chromatography (LC) procedure is described for the determination of insulin in biological matrices. The total procedure consists of two SPEs in series, followed by RP-LC separation. During the first SPE a strong anion-exchange (SAX) cartridge (ISOLUTE, 40-90 microm, 10 x 4 mm i.d.) is used, followed by a RP-cartridge (Luna C(8), 4 x 2.0 mm i.d.). The second SPE cartridge contains the same material as the LC column and is used to transfer the sample from the SAX cartridge to the LC column. The developed system can detect 100 nmol/l insulin in aqueous samples and 200 nmol/l insulin in spiked plasma samples using UV. When electrospray ionization (ESI) mass spectrometry (MS), was coupled with the developed system, the LODs were lowered by a factor two to 50 and 100 nmol/l for aqueous and spiked plasma samples, respectively. PMID- 12763538 TI - High-performance liquid chromatographic method for the quantification of anthranilic and 3-hydroxyanthranilic acid in rat brain dialysate. AB - Anthranilic acid (ANA) and 3-hydroxyanthranilic acid (3-HANA) have attracted considerable attention as two of the L-tryptophan kynurenine pathway metabolites in the central nervous system. In this study, a highly sensitive and accurate method for the quantification of ANA and 3-HANA has been developed using reversed phase high performance liquid chromatography (HPLC) with fluorimetric detection. The HPLC assay was carried out using a C(18) column (5 microm, 250 x 4.6 mm i.d.). The mobile phase consisted of a mixture of 25 mM sodium/acetic acid buffer (pH 5.5) and methanol (90:10 v/v). Fluorimetric detection at lambda(ex)=316 nm and lambda(em)=420 nm was used. The assay was applied to the measurement of ANA and 3-HANA acid in rat brain dialysate following administration of L-tryptophan or L-kynurenine. 3-HANA and ANA levels were progressively increased during 90 min following administration of L-tryptophan, then decreased progressively to basal levels. 3-HANA levels were significantly higher than ANA levels after L kynurenine administration. These findings suggest that the assay developed should provide an improved means for investigation of neurobiology of kynurenine pathway. PMID- 12763541 TI - Simultaneous determination of sildenafil and its active metabolite UK-103,320 in human plasma using liquid chromatography-tandem mass spectrometry. AB - A liquid chromatography-tandem mass spectrometric method for the simultaneous determination of sildenafil and its active N-demethylated metabolite, UK-103,320 in human plasma was developed. Sildenafil, UK-103,320 and the internal standard (DA-8159) were extracted from human plasma with dichloromethane at basic pH. A reverse-phase LC separation was performed on Luna phenylhexyl column with the mixture of acetonitrile-ammonium formate (10 mM, pH 6.0) (60:40, v/v) as mobile phase. The detection of analytes was performed using an electrospray ionization tandem mass spectrometry in the multiple reaction-monitoring mode. The lower limits of quantification for sildenafil and UK-103,320 were 2.0 ng/ml. The method showed a satisfactory sensitivity, precision, accuracy, recovery and selectivity. PMID- 12763542 TI - Analysis of nabumetone in human plasma by HPLC. Application to single dose pharmacokinetic studies. AB - A simple and sensitive high performance liquid chromatography method for the determination of nabumetone in human plasma is described. The procedure involves liquid-liquid extraction with ethyl acetate and reversed-phase chromatography with fluorimetric detection (excitation 230 nm, emission 356 nm). The chromatographic conditions and the extraction procedure gave a clean chromatogram for the compound. The limit of quantitation was established as 0.313 ng/ml and the calibration curve was linear up to 20 ng/ml. The within-day and between-day relative standard deviations were less than 10% and the accuracy of the assay was in the range of 99-104%. The suitability of the method is shown for pharmacokinetic studies. PMID- 12763543 TI - Quantitative determination of sufentanil in human plasma by liquid chromatography tandem mass spectrometry. AB - A sensitive and specific method for the quantification of sufentanil in human plasma by liquid chromatography coupled with tandem mass spectrometry has been developed. Fentanyl was used as the internal standard. Rapid sample preparation involved purification on a C(18) solid-phase extraction column. Chromatographic separation of the analytes was obtained using an RP-C(18) mu-HPLC column. LC-MS MS detection was performed by atmospheric pressure ionisation (API) source equipped with an ionspray (IS) interface operating in the positive ion mode. For unambiguous substance confirmation, three analyte precursor-product ion combinations were monitored during multiple reaction monitoring (MRM) LC-MS-MS analysis. The method's performance characteristics were evaluated in blank and spiked control plasma samples. Overall accuracy (relative error, R.E., %), repeatability (relative standard deviations, R.S.D., %) and within-laboratory reproducibility (R.S.D., %) ranged from -9.28 to -2.71%, from 6.42 to 2.82% and from 13.52 to 6.06%, respectively, for sufentanil. The limit of quantification for sufentanil in human plasma samples was 0.3 ng ml(-1). Due to its high sensitivity and specificity, the method was successfully employed for sufentanil determination in maternal plasma samples collected immediately before epidural administration of a single sufentanil dose to women in labour, 20 min after drug administration, and at birth in arterial and venous umbilical cord plasma samples from the newborn babies. Research is in progress to adopt the method for performance of complete pharmacokinetic studies of sufentanil in human plasma. PMID- 12763544 TI - Analysis of albendazole metabolites by electrospray LC-MS/MS as a probe to elucidate electro-oxidation mechanism of albendazole. AB - The electrochemical oxidation of albendazole was accomplished by controlled potential electrolysis technique. The oxidation was carried out in different pH solutions and yields the same products obtained by in vivo and in vitro metabolism, i.e. albendazole sulfoxide and albendazole sulfone. The identification of albendazole oxidation products was carried out by LC-MS/MS. PMID- 12763545 TI - Development and validation of a liquid chromatographic method for the determination of amlodipine residues on manufacturing equipment surfaces. AB - In the pharmaceutical industry, an important step consists in the removal of possible drug residues from the involved equipment and areas. The cleaning procedures must be validated and the methods to determine trace amounts of drugs have therefore to be considered with special attention. A high performance liquid chromatographic method for the determination of amlodipine residues in swab samples was developed and validated in order to control a cleaning procedure. The swabbing procedure was optimized in order to obtain a suitable recovery of amlodipine from stainless steel. A mean recovery close to 90% was obtained when two swabs moistened with methanol were used. The residual amlodipine was chromatographed at 25 degrees C in the isocratic mode on a RP-18 stationary phase using a mobile phase consisting of acetonitrile, methanol and pH 3.0 triethylamine solution (15:35:50 v/v/v). UV detection was performed at 237 nm. The method was shown to be selective and linear into the concentration range varying from 0.39 to 1.56 microg/ml. Accuracy and precision of the method were also studied. The limits of detection and quantitation were evaluated to be 0.02 and 0.08 microg/ml, respectively. The stability of amlodipine at different steps of the sampling procedure and the precision of the swabbing procedure were also investigated. PMID- 12763546 TI - Anodic polarographic determination of ciclopirox olamine in pure and certain pharmaceutical preparations. AB - The anodic polarographic behavior of ciclopirox have been studied in Britton Robinson buffer (BRb) over the pH range 6-11. In BRb of pH 7 a well defined anodic wave was produced with diffusion current constant (Id) of 4.86+/-0.048 (n=6) using DC(t) mode. Adopting both of direct current (DC(t)) and differential pulse polarographic (DPP) modes, the current-concentration relationship was found to be rectilinear over the range 4 to 24 and 2 to 12 microg ml(-1) respectively, with minimum detectability (S/N=2) of 0.2 microg ml(-1) (1 x 10(-6) M) using the DPP mode. The average percent recovery was favourably compared to a reference method, with a satisfactory standard deviation, the proposed method was further applied to the determination of ciclopirox olamine in certain pharmaceutical preparations including lotion and cream. The average percentage recoveries for lotion were 100.06+/-0.94 and 100.06+/-1.08 using DC(t) and DPP modes respectively, and for cream were 100.17+/-0.64 and 100.34+/-1.28 using DC(t) and DPP modes, respectively. A pathway for the electrode reaction was postulated. PMID- 12763547 TI - Application of LC-MS/MS for the identification of a polar impurity in mosapride, a gastroprokinetic drug. AB - In the impurity profile of mosapride a polar impurity (0.1%) was detected in HPLC with respect to mosapride. Based on the mass spectral data obtained by LC-MS/MS analysis this impurity structure was characterised as 4-amino-5-chloro-2-ethoxy-N [[(4-benzyl)-2-morphinyl] methyl] benzamide. It is interesting to note that this impurity is potent analogue of mosapride, which will have much higher gastroprokinetic activity than metoclopramide. This impurity was synthesised from an unambiguous route and confirmed the structure by collecting various spectral data and the formation is discussed. To our knowledge this compound was not reported as process impurity elsewhere. PMID- 12763548 TI - Determination of diphenytriazol (DL111-IT) and its related impurities by RP-HPLC with DAD. AB - An analytical method was developed for determining diphenytriazol and its related impurities in oil injection by using RP-HPLC with DAD and diazepam as internal standard. The C(18) column was used as analytical column. Mobile phase consisted of methanol-potassium dihydrogen phosphate solutions (10 mmol l(-1), pH 7.5) (7:3, v/v). The standard curve was linear in the concentration range from 2 to 100 microg ml(-1) for diphenytriazol. The analytical method afforded average recoveries of 100.3+/-1.9% (n=9) and the relative standard deviation (RSD) was less than 2% for within-day and between-day precision. The limit of detection and of quantitation for the assay were 15 and 40 ng, respectively. The method was simple, accurate and allowed to be used as analytical method for the routine quality control of diphenytriazol injection. Diphenytriazol injection showed a high stability to the heat (60 degrees C) and the light (4000 lx). PMID- 12763549 TI - Liquid chromatographic determination of unbound flecainide in therapeutic drug monitoring. AB - An assay method was developed for determining unbound flecainide in serum by reversed phase-high performance liquid chromatography (HPLC). Serum water including unbound flecainide was separated by ultrafiltration of the serum sample and subjected to C(18)-cartridge extraction followed by HPLC analysis. The recovery of flecainide from serum water was greater than 93%. The coefficient variations for intra- and inter-day assay of flecainide were smaller than 2.4 and 3.7%, respectively. We applied the method to determining unbound flecainide in serum samples collected from 20 patients receiving oral flecainide (150-300 mg/day) for tachyarrhythmia. Total and unbound concentrations for serum flecainide were 403.5+/-200.8 ng/ml and 180.2+/-95.0 ng/ml, respectively. Linear relationship was observed between total and unbound concentrations (r=0.978, P<0.0001). Percent unbound (44.3+/-5.7%) determined in the present study agreed with the reported values. The percentage unbound tended to increase in the samples with lower alpha(1)-acid glycoprotein (<60 mg/dl). The assay method can be applied to routine determination of unbound flecainide in therapeutic drug monitoring. PMID- 12763550 TI - Determination of cetylpyridinium chloride and tetracaine hydrochloride in buccal tablets by RP-HPLC. AB - The HPLC method for simultaneous determination of cetylpyridinium chloride (CPC), tetracaine hydrochloride (TTC) in Xipiluan buccal tablets was developed and validated. The HPLC method was performed on a CN column (150 x 4.6 mm i.d., 5 microm particle size); the mobile phase was methanol-tetramethylammonium hydroxide (20 mM)-potassium dihydrogen phosphate (3 mM) (90:10:3, v/v/v) (pH* 5.0), pumped at a flow rate 1.5 ml min(-1). The UV detector was set at 230 nm. The retention time for CPC and TTC was 3.52 and 3.10 min, respectively. Calibration curves were linear (r=0.9999, n=6) in the range of 5-2000 microg ml( 1) for CPC and 1-500 microg ml(-1) for TTC. Limit of detection and quantitation for CPC was 0.033 and 0.11 microg ml(-1), for TTC were 0.0056 and 0.019 microg ml(-1). The R.S.D. of repeatability and intermediate precision for CPC and TTC were less than 2.0%. PMID- 12763551 TI - High-performance liquid chromatographic analysis of nitrendipine in human plasma using ultraviolet detection and single-step solid-phase sample preparation. AB - A high-performance liquid chromatographic (HPLC) method was developed for the determination of nitrendipine in human plasma using solid-phase extraction (SPE) and ultraviolet detection. A 30-microl aliquot of methanol (containing 2 microg/ml of the internal standard, nimodipine) was added to a 1-ml aliquot of biological sample. After vortex-mixing, the mixture was loaded on C(18) SPE cartridge which was conditioned with methanol and distilled water. After washing with distilled water, the SPE cartridge was eluted with 1-ml aliquot of diethyl ether. The organic phase was collected and evaporated under nitrogen gas. The residue was then reconstituted with a 100 microl aliquot of mobile phase, and a 50 microl aliquot was injected onto the C(18) reverse-phased column. The mobile phase, 10 mM phosphate buffer (pH 4.5):acetonitrile (50:50, v/v), was run at a flow rate of 1.2 ml/min. The column effluent was monitored using ultraviolet detector at 238 nm. The retention times for nitrendipine and the internal standard were approximately 10.1 and 12.6 min, respectively. The detection limit of nitrendipine in human plasma was 2.0 ng/ml. The coefficients of variation (CV) of the assay were below 16.5% for human plasma, and no interferences from endogenous substances were found. This specific, accurate and precise assay was useful in the study for the pharmacokinetic characteristics of nitrendipine. PMID- 12763552 TI - Biological replacement heart valves. Identification and evaluation. PMID- 12763554 TI - Surgical pathology of subaortic septal myectomy associated with hypertrophic cardiomyopathy. A study of 204 cases (1996-2000). AB - BACKGROUND: No large surgical series have qualitatively examined all histopathologic features of hypertrophic cardiomyopathy (HCM). METHODS: Medical records and microscopic slides were reviewed from 204 patients undergoing septal myectomy for HCM at Mayo Clinic Rochester. RESULTS: The 108 females and 96 males (1-86 years old; mean, 48) included 133 patients <60 years old (Group 1) and 71 patients > or =60 years (Group 2). Clinical features more prevalent in Group 2 than in Group 1 included female gender (68% vs. 45%; P=.003), aortic valve disease (31% vs. 12%; P=.01) and severe coronary atherosclerosis (43% vs. 9%; P=.001). Overall, microscopic abnormalities included myocyte hypertrophy (100%), endocardial (96%) and myocardial (93%) fibrosis, myocyte disarray (79%) and vacuolization (60%), endocardial inflammation (48%), arterial thickening (46%), dilated venules (28%), arterial dysplasia (16%), left bundle branch tissue (12%), infarction (2%), endocardial (1%) and myocardial (<1%) calcium, and amyloid (1%; senile type in all three). Lesions more prevalent in Group 1 than Group 2 were vacuolization (68% vs. 45%; P=.002), disarray (87% vs. 65%; P=.0003) and dilated venules (33% vs. 18%; P=.02). In contrast, lesions more frequent in Group 2 than in Group 1 included left bundle branch (20% vs. 8%; P=.02) and amyloid and endocardial calcium (4% vs. 0%; P=.04, each). CONCLUSIONS: Among patients undergoing septal myectomy for HCM, 53% were women and 35% were >/=60 years old. The most common microscopic features were hypertrophy, disarray, fibrosis, inflammation and vascular alterations. Disarray cannot be used as a morphologic hallmark for HCM in small surgical myectomy specimens because it was absent in 21% of the patients. Because amyloid unexpectedly affected three elderly patients, routine amyloid staining is recommended for patients >/=65 years old. PMID- 12763555 TI - Endodermal heterotopia of the atrioventricular node associated with transposition of the great arteries. AB - A 13-year-old girl with surgically corrected complete transposition of the great arteries died suddenly while walking to school. Postmortem examination revealed endodermal heterotopia of the atrioventricular node, a condition not previously associated with transposition of the great arteries. New immunohistochemical observations are described, and the embryogenesis of the condition and its association with complete transposition are discussed. PMID- 12763553 TI - Postmortem cardiomegaly and echocardiographic measurements of left ventricular size and function in children infected with the human immunodeficiency virus. The Prospective P2C2 HIV Multicenter Study. AB - BACKGROUND: Cardiomegaly, seen postmortem in over 50% of HIV-infected children, may occur in the absence of clinical or histopathologic cardiac disease. Premortem echocardiography has also demonstrated clinical and subclinical cardiac disease, including increased left ventricular mass (LVM) and functional abnormalities. No studies have compared these echocardiographic measures of heart size and function with postmortem findings in this population. We sought to determine the postmortem prevalence, clinicopathologic relationships and importance of cardiomegaly in HIV-infected children. METHODS: We reviewed clinical and postmortem cardiac findings in 30 HIV-infected children who were part of the Prospective P(2)C(2) HIV Study. Postmortem heart weight was compared with clinical measures of heart size and function, with cardiac pathology and with clinical measures reflecting chronic effects of HIV disease. RESULTS: Postmortem cardiomegaly (heart weight z score >/=2) was identified in 53% of the children. Children with cardiomegaly had increased LVM, increased heart rate, more frequent clinical chronic heart disease and a higher prevalence of postmortem pericardial effusions compared to children without cardiomegaly (P0.05) in brain % water content and extracellular fluid [14C]sucrose volume of distribution between SHR and WKY rats. However, choroid plexus showed greater % water content in SHR (85.7+/-1.9%) compared to the WKY rats (81.5+/-1.7%). These results suggest that chronic hypertension in SHR may cause more pronounced defects in the integrity of the blood-CSF barrier than in the BBB. PMID- 12763607 TI - Rapid induction of Arc is observed in the granule cell dendrites in the accessory olfactory bulb after mating. AB - The activity-regulated cytoskeleton-associated protein (Arc), encoded by the immediate early gene arc, is enriched in the brain and is hypothesized to play a role in the activity-dependent neuronal plasticity in the hippocampus. In the present study, the time course of Arc expression during the post-mating period was determined immunocytochemically, and the localization of Arc in the neurons in the accessory olfactory bulb (AOB) of female mice after mating was analyzed using immunocytochemical electron microscopy. Transient increases in the number of Arc-immunoreactive cells were observed in the glomerular, mitral/tufted cell and granule cell layers of the AOB after mating. In particular, the increase in the granule cell layer was remarkable, and larger than the increases in the other layers. In addition, electron microscopic observation revealed that Arc immunoreactivity was in the dendrites of the granule cells 1.5 h after mating. These results indicate that expression of Arc protein is induced rapidly and transiently in granule cell dendrites after mating. It is postulated that Arc protein has a role in the neuronal plasticity of the AOB after mating. PMID- 12763608 TI - GABA(A) alpha1 and alpha2 receptor subunit expression in rostral ventrolateral medulla in nonpregnant and pregnant rats. AB - Pregnancy results in attenuated baroreflex mediated sympathoexcitatory responses which may be due to potentiation of gamma-aminobutyric acid (GABA) inhibition in the rostral ventrolateral medulla (RVLM). The major metabolite of progesterone, 3alpha-hydroxy-dihydroprogesterone (3alpha-OH-DHP), which is elevated in pregnancy, is a potent neurosteroid positive modulator of GABA(A) receptors, and sensitivity of GABA(A) receptors to 3alpha-OH-DHP is dependent on the receptor subunit composition. The purpose of this study was to evaluate the GABA(A) alpha(1) and alpha(2) receptor subunit mRNA and protein expression in the RVLM of nonpregnant and late term pregnant rats. Micropunches of RVLM were collected from nonpregnant and late term pregnant rats and the expression levels of GABA(A) alpha(1) and alpha(2) receptor subunits were analyzed using quantitative competitive reverse transcriptase polymerase chain reaction (RT-PCR) and immunoblot techniques. The competitive RT-PCR analysis allows comparison of expression levels between different mRNA, and the mRNA expression level of GABA(A) alpha(1) was several hundred fold greater than GABA(A) alpha(2) in both groups. However, this relative distribution of GABA(A) alpha(1) and alpha(2) receptor subunits protein or mRNA expression was not altered in late term pregnant compared to nonpregnant rats. These data demonstrate, that within the RVLM of both nonpregnant and late term pregnant rats, the relative expression levels of GABA(A) alpha(1,2) receptor subunits favor GABA(A) receptors susceptible to positive modulation by progesterone metabolites. PMID- 12763609 TI - Differential involvement of hippocampal and amygdalar NMDA receptors in contextual and aversive aspects of inhibitory avoidance memory in rats. AB - Adult male rats bilaterally implanted with guide canullae aimed either at the dorsal hippocampus (dHIP) or the basolateral nucleus of the amygdala (BLA) were trained in a step-down inhibitory avoidance task (IA) and tested for retention 24 h after training. Immediately after training, animals were given a bilateral infusion of the N-methyl-D-aspartate (NMDA) glutamate receptor antagonist D,L-2 amino-5-phosphonopentanoic acid (AP5) (5.0 microg) into the dHIP or the BLA. Both intrahippocampal and intraamygdala infusions of AP5 blocked IA retention. Preexposure to the training box, but not to a different environment 24 h prior to training prevented the impairing effect of intrahippocampal infusion of AP5 on retention. Preexposure did not affect the retention impairment induced by intraamygdala infusion of AP5. These data suggest that hippocampal NMDA receptors might be involved in the contextual and spatial aspects, while amygdalar NMDA receptors might be involved in the aversive aspects of memory for IA. PMID- 12763610 TI - Rapid reduction of ATP synthesis and lack of free radical formation by MPP+ in rat brain synaptosomes and mitochondria. AB - MPTP is a neurotoxin thought to damage dopaminergic neurons through free radical formation. MPTP is metabolized in the brain to MPP(+), which is taken up into dopaminergic neurons via the dopamine transporter and assumed to impair mitochondrial function. We used striatal synaptosomes and telencephalic mitochondria to further investigate MPP(+) mechanism of action. For comparison, the respiratory toxins FCCP, a cyanide analog that uncouples mitochondrial ATP production, and rotenone, a NADH dehydrogenase inhibitor, were also tested. FCCP, MPP(+) and rotenone caused a rapid but stable decrease in [3H]dopamine (DA) uptake by striatal synaptosomes. Two free radical scavengers, the salen-manganese complex EUK-134, and the spin trap s-PBN, did not prevent MPP(+)-induced decrease in DA uptake. However, addition of ATP during synaptosome preparation resulted in partial recovery of MPP(+)-induced [3H]DA uptake decrease. Generation of oxygen free radicals by treatment of telencephalic mitochondria with MPP(+), FCCP, or rotenone, was evaluated by measuring DCF fluorescence, while light emission by the luciferin-luciferase complex was used to determine ATP levels. MPP(+), unlike rotenone, did not produce oxygen free radicals, but rather blocked ATP production in mitochondria, as did FCCP and rotenone. Taken together, these results suggest that MPP(+) toxicity, at least during its initial stages, is primarily due to a decrease in ATP synthesis by mitochondria and not to free radical formation. PMID- 12763611 TI - Evidence for the involvement of a spinal pattern generator in the control of the genital motor pattern of ejaculation. AB - One of the hypotheses to explain the neural mechanisms underlying rhythmic behaviours suggests that the central nervous system has the intrinsic capacity to produce repetitive, rhythmic output to the muscles involved in the response by means of a neuronal circuit named central pattern generator (CPG). The occurrence of rhythmic motor patterns during ejaculatory behaviour in mammals, which includes the genital motor pattern, has been shown. A CPG might regulate the timing of the repetitive muscular responses that constitute the ejaculatory motor pattern. The objective of the present study was to evidence that a CPG at a spinal level is involved in the expression and pacing of the rhythmic motor pattern generated during ejaculation. To this purpose we used the genital reflex as a model system. Following the general principles for the study of rhythmic motor patterns, the data obtained in the present series of experiments document that: (1) a rhythmic muscular response, the genital motor pattern, is registered during the ejaculatory event (expulsion of the urethral contents); (2) this ejaculatory motor response has similar EMG characteristics in intact and in spinal urethane-anaesthetised male rats; (3) interruption of the afferent inflow (deafferentation) does not disrupt the expression of the ejaculatory motor train; (4) a change in the stimulation interval does not alter the intrinsic pacing of the ejaculatory-like response; and (5) fictive ejaculation can be induced by pharmacological means. Together, this evidence supports the notion that a CPG produces the rhythmic ejaculatory motor pattern registered during fictive ejaculation. PMID- 12763612 TI - Valproic acid enhances axonal regeneration and recovery of motor function after sciatic nerve axotomy in adult rats. AB - It has recently been demonstrated that valproic acid (VPA) robustly promotes neurite outgrowth, activates the extracellular signal regulated kinase pathway, and increases growth cone-associated protein 43 and bcl-2 levels in cultured human neuroblastoma SH-SY5Y cells. We hypothesized that VPA could also enhance peripheral nerve regeneration in adult animals. To test this hypothesis, we examined the effects of VPA (300 mg/kg daily for 16 weeks) on sciatic axonal regeneration following single or conditional axotomies in rats. The results showed that in VPA-treated rats there was a significant increase in the total numbers of regenerated myelinated nerve fibers and reinnervated muscle fibers in comparison with those rats not treated with VPA. As measured by sciatic function index and toe spread index, the motor function of the reinnervated hind limbs of rats receiving single axotomy without VPA treatment significantly improved at week 8 and reached plateau levels at about week 11, whereas the motor function of the reinnervated hind limbs of rats receiving single axotomy plus VPA and rats receiving conditional axotomy with or without VPA treatment significantly improved at week 4 and reached plateau levels at about week 8; there was no significant difference of the motor function among the three later groups. The results demonstrated that VPA is able to enhance sciatic nerve regeneration and recovery of motor function in adult rats, suggesting the potential clinical application of VPA for the treatment of peripheral nerve injury in humans. PMID- 12763613 TI - Adenosine-mediated synaptic depression and EPSP/spike dissociation following high potassium-induced depolarization in rat hippocampal slices. AB - Simultaneous recordings of orthodromic PS, fEPSP and antidromic PS revealed EPSP/spike (E-S) dissociation, indicating a conversion of input/output relations from early and brief excitability to a late and prolonged depression during the recovery from depolarization induced by high levels of potassium. E-S potentiation was partially attenuated by pre-treating the slices with BAPTA-AM and lidocaine and totally eliminated by a submaximal concentration of muscimol. The time lag for recovery was decreased by the GABA(A) antagonist and completely eliminated by the A(1) antagonist. From these observations, we conclude that Ca(2+) dependent inhibitory suppression is the main cause of a brief period of E S potentiation, and accumulation of adenosine is the mechanism responsible for prolonged depression of synaptic transmission PMID- 12763614 TI - Severe brain hypothermia as a factor underlying behavioral immobility during cold water forced swim. AB - Behavioral immobility during forced swim is usually considered a consequence of inescapable stress, and is used to screen antidepressant drugs. However, immobility in this test may also result from inhibition of neural functions because of brain hypothermia due to body cooling. To explore this possibility, we measured brain temperature dynamics during a 10-min forced swim in cold (25 degrees C) and warm (37 degrees C) water and correlated brain temperatures with behavioral changes. Cold water forced swim resulted in significant brain hypothermia (-6-7 degrees C) and immobility, while no immobility was observed during warm water forced swim, when brain temperature transiently increased (0.5 degrees C) then decreased below baseline in the post-swim period. These data suggest that immobility, which rapidly develops during forced swim in cold water, may result from dramatic inhibition of neural functions because of severe brain hypothermia. PMID- 12763616 TI - Region-specific dysregulation of allopregnanolone brain content in the olfactory bulbectomized rat model of depression. AB - Allopregnanolone (ALLO) is one of the most potent positive endogenous allosteric modulators of the type A gamma-aminobutyric acid (GABA(A)) receptors. While the robust anxiolytic profile of ALLO has been extensively characterized in rodents and its antidepressant-like effect was recently demonstrated in mice, there have been only few reports on alterations of brain ALLO levels in putative animal models of depression and anxiety. Removal of the olfactory bulbs of rats produces one of the most predictive animal models with which to screen for drugs with potential antidepressant activity following repeated treatment. We therefore investigated whether the olfactory bulbectomized (OB) rat model of depression may be associated with alterations of ALLO levels in whole brain tissue and in different brain regions. We determined ALLO levels in whole brain, amygdala, frontal cortex, hippocampus, and whole cerebral cortex of OB or sham-operated rats at 7, 14, or 28 days following bulbectomy or sham surgery. We observed a significant increase of whole brain ALLO content at 7 and 28 days post-surgery in the OB rats. At days 7 and 14 following olfactory bulb removal, ALLO levels were significantly decreased in amygdala and frontal cortex and significantly increased in whole cerebral cortex. In the hippocampus we observed only a tendency for decreased ALLO levels at day 14. Our data indicates a strong region specific dysregulation of ALLO homeostasis in brains of OB rats which may contribute to the formation of the bulbectomy syndrome via a sustained reduction in physiological GABA-ergic tone in amygdala and frontal cortex. PMID- 12763617 TI - GFAP and nuclear lamins share an epitope recognized by monoclonal antibody J1-31. AB - Monoclonal antibody J1-31 was raised against plaque materials taken from brains of patients who had suffered from multiple sclerosis (MS). Preliminary characterization of the antigen revealed it to be a protein of M(w) 68-70 kDa with both a cytoplasmic and nuclear localization. Here we report the results of isolation and peptide sequencing of the antigen from human brains, and immunocytochemical analysis of the antigen in F98 glioma cells. Purification and peptide sequencing indicate that the antibody recognizes a form of glial fibrillary acidic protein, possibly a phosphorylated variant. However, confocal immunocytochemistry and western analysis of F98 glioma cells raise the possibility that it also recognizes a phosphorylated epitope found in nuclear lamins. Analysis of the expression of the J1-31 epitope in F98 cells with respect to time in culture, cell density, and DNA synthesis showed a developmental relationship: cells that were engaged in rapid growth and DNA synthesis exhibited strong J1-31 staining in nuclei, whereas quiescent cells did not. We conclude that mAB J1-31 remains a useful antibody for studying multiple sclerosis, and is likely to prove useful in studies of the dynamics of nuclear lamins, particularly in models for wound-healing. PMID- 12763618 TI - The effects of antidepressant treatment on serotonergic and dopaminergic systems in Fawn-Hooded rats: a quantitative autoradiography study. AB - Fawn-Hooded (FH) rats exhibit a phenotype including depressive behaviour and high alcohol preference, and as such tricyclic antidepressants and selective serotonin reuptake inhibitors (SSRIs) reduce alcohol consumption in this rat strain [Psychiatr. Genet. 12 (2002) 1-16]. However, the neurochemical effects of these antidepressants on monoamine systems in the brain, especially in mesolimbic areas have not been studied in FH rats. Therefore, the present study investigated neurochemical effects of subchronic treatment (10 days) with desipramine (DMI) and sertraline on several neurochemical markers of serotonin and dopamine systems. Binding to these markers including dopamine transporters (DATs), 5-HT transporters (SERTs), 5-HT(1A)- and 5-HT(2A)-receptors in rat brain sections was performed by quantitative autoradiography. The findings from the present study revealed that DMI and sertraline exhibited differential effects on SERTs and DATs in FH rat brain. For example, DMI caused a dramatic up-regulation of DATs whereas sertraline had no effect on DAT binding. In addition, both antidepressants showed some common and some differential effects on the binding to 5-HT(1A)- and 5 HT(2A)-receptors dependent upon region. These data demonstrate that DMI and sertraline differentially effect serotonergic and dopaminergic systems in mesolimbic regions in FH rats, suggesting that there may be different neurochemical mechanisms underlying their efficacy to reduce ethanol consumption in this animal model. PMID- 12763619 TI - Sulpiride injections into the medial septum reverse the influence of intra-medial septum injection of L-arginine on expression of place conditioning-induced by morphine in rats. AB - Effects of intra-medial septum injections of L-arginine, a precursor of nitric oxide, N(G)-nitro-L-arginine methyl ester (L-NAME), an inhibitor of nitric oxide synthase, and sulpiride, a selective antagonist of dopamine D2 receptor on morphine-induced conditioned place preference (CPP) in male Wistar rats were examined. Using a 3-day schedule of conditioning, morphine (0.5-7.5 mg/kg, s.c.) produced a significant place preference in a dose-dependent manner. The maximum response was observed with 5.0 mg/kg of opioid. Sulpiride (0.3, 1.0 and 3.0 microg/rat), but not L-arginine (0.3, 1.0 and 3.0 microg/rat) or L-NAME (0.3, 1.0 and 3.0 microg/rat), in combination with morphine (5.0 mg/kg), during conditioning, significantly altered morphine-induced CPP. Single doses (0.3, 1.0 and 3.0 microg/rat) of either L-arginine or L-NAME, during conditioning, did not induce CPP. Sulpiride at 0.3-3.0 microg/rat, intra-medial septum, during conditioning, produced a significant conditioned place aversion. Intra-medial septum injections of L-arginine but not L-NAME or sulpiride, 1-2 min before testing, increased the expression of morphine-induced CPP. The administration of sulpiride (0.3, 1.0 and 3.0 microg/rat), but not L-NAME (0.3, 1.0 and 3.0 microg/rat), 1-2 min before the injection of L-arginine (0.3 microg/rat) on day of test, significantly attenuated the response to L-arginine. L-Arginine (0.3-3.0 microg/rat), during conditioning, showed a statistically significant increase in locomotor activity compared with that to control group. Moreover, sulpiride decreased locomotion by itself or in combination with morphine during conditioning and on the test day of morphine CPP. It can be concluded that L arginine, a precursor of nitric oxide, in the rat median septum may play a role in expression of morphine conditioning due to dopamine release in this area. PMID- 12763620 TI - 17beta-estradiol inhibits excitatory amino acid-induced activity of neurons of the nucleus tractus solitarius. AB - The effects of 17beta-estradiol (17betaE2) on spontaneous and excitatory amino acid (EAA) induced nucleus tractus solitarius (NTS) neuronal activity were investigated by electrophysiological and immunohistochemical experiments in ovariectomized female Sprague-Dawley rats. Out of 62 NTS neurons tested, 42 were inhibited (68%) following iontophoretic application of 17betaE2 in a current dependent manner. The averaged firing rate decreased from 3.06+/-0.40 to 0.78+/ 0.17 Hz. The inhibitory responses were rapid in onset (within 1 min) and variable in duration (2-4 min). The inhibitory effects of 17betaE2 were blocked by simultaneously applied 17betaE2 antagonist ICI182,780, but not by GABA antagonist, bicuculline and phaclofen. L-Glutamate, AMPA or NMDA enhanced the activity of 71, 73 or 69% of NTS cells tested, respectively. The excitatory effects of EAA were significantly inhibited in the presence of 17betaE2. Fluorescent immunohistochemistry revealed that all subnuclei of the NTS contained high levels of estrogen receptors (ERs) immunoreactivity. These results suggest that 17betaE2 inhibits spontaneous and EAA-induced NTS neuronal activity through 17betaE2 activation of ERs. PMID- 12763621 TI - Retinal expression of clusterin in the streptozotocin-induced diabetic rat. AB - To assess the possible relevance of clusterin in the pathophysiology of retinopathy associated with diabetes mellitus, streptozotocin-induced diabetic rats were studied. Clusterin expression was measured in both normal and streptozotocin-induced diabetic rat retinas using Northern blotting, reverse transcription polymerase chain reaction, immunohistochemistry and Western blot analysis. The results show increased clusterin protein level and its mRNA expression 6 weeks after induction of diabetes. Clusterin was localized to the inner nuclear and ganglion cell layers of both normal and diabetic rat retinas. These data show that diabetes affects the expression of clusterin in the retina and may reflect a diabetes-induced damage and/or alterations of neural structures resulting in diabetic retinopathy. PMID- 12763622 TI - Postsynaptic modulation of AMPA- and NMDA-receptor currents by Group III metabotropic glutamate receptors in rat nucleus accumbens. AB - Whole cell patch clamp recordings from rat nucleus accumbens neurons were made in order to study the effect of metabotropic glutamate receptors and dopamine on postsynaptic glutamate receptor mediated currents. AMPA- and NMDA-R currents were evoked by flash photolysis of caged glutamate, while spike-dependent release of neurotransmitters was prevented by adding tetrodotoxin and bicuculline to the bath solution. Spontaneous potentiation of NMDA- but not AMPA-R current was observed in the early phase of stimulation, followed by depotentiation and subsequent stabilization. The Group III metabotropic glutamate receptor antagonist MAP4 induced a transient potentiation of both AMPA- and NMDA-R current amplitudes, without affecting rise times and decay time constants. In contrast, the Group I-II metabotropic glutamate receptor antagonist MCPG and the neurotransmitter dopamine did not exert significant effects on either AMPA- or NMDA-R currents. These data suggest that at least one of the Group III subtypes is located postsynaptically in the nucleus accumbens and is able to dampen the activity of ionotropic glutamatergic receptors. In contrast, our results do not support a modulation of postsynaptic AMPA- and NMDA-R currents by Group I/II metabotropic glutamate receptors or dopamine. Modulation of both AMPA- and NMDA-R currents in the nucleus accumbens is likely to play a major role in setting the cellular excitability in response to behaviourally relevant limbic inputs, and in regulating the plasticity of these responses. PMID- 12763623 TI - TNFalpha-stimulation of cFos-activation of neurons in the solitary nucleus is suppressed by TNFR:Fc adsorbant construct in the dorsal vagal complex. AB - The cytokine tumor necrosis factor alpha (TNF(alpha)) may act within the neural circuitry of the medullary dorsal vagal complex (DVC) to affect changes in gastric function such as gastric stasis, loss of appetite, nausea, and vomiting. The definitive demonstration that endogenously generated TNF(alpha) is acting within the DVC circuitry to affect these changes has been impeded by the lack of an antagonist for TNF(alpha). The present studies used localized central nervous system microinjections of the TNF-adsorbant construct (TNFR:Fc) to specifically neutralize the ability of endogenously produced TNF(alpha) to activate NST neurons. Our studies reveal that TNFR:Fc suppresses induction of cFos normally evoked by TNF(alpha). These results validate our hypothesis that circulating TNF(alpha) may act directly within the DVC to affect gastric function in a variety of pathophysiological states. PMID- 12763624 TI - Actions of oxytocin and interactions with glutamate on spontaneous and evoked dorsal spinal cord neuronal activities. AB - Among the numerous pain control mechanisms that have been proposed, those acting at the spinal cord have been broadly studied, but little is known about how neuropeptides originating in supraspinal structures may relate to pain and analgesic mechanisms. Oxytocin (OT), in addition to its well known hormonal action, produces neuronal effects in various regions of the central nervous system. Indeed, some parvocellular neurons in the hypothalamic paraventricular nucleus (PVN) are oxytocinergic and project to the caudal part of the brain and the spinal cord. Moreover, the rat spinal cord shows a good overlap between the oxytocinergic hypothalamo-spinal neuron projections and the distribution of OT binding sites. However, the physiological significance of these binding sites is largely unknown. Extracellular unit activity of spinal cord neurons was recorded at the T13-L1 levels in male rats anesthetized with halotane. Somatic stimulation was applied to the inner and outer thigh of the ipsilateral hindpaw, and glutamate (GLU) and OT were locally delivered by pressure using pipettes coupled to recording electrodes. Our results show that spinal cord neurons, mainly located in the dorsal horn, in the intermediolateral cell column (IML) and in the intermediomedial gray matter (IMM), respond to the application of OT (71.5%) with activation (48%) or inhibition (52%). In some cases, opposite OT effects were observed during simultaneous recordings of two cells, suggesting OT activation of an inhibitory interneuron followed by the inhibition of the second recorded neuron. Increases in neuronal firing rate produced by GLU could be blocked by prior OT application. Finally, OT could reduce or partially block the responses to tactile and nociceptive somatic stimulation. We found that spinal cord neurons are sensitive to OT indicating that OT binding sites are functionally active. OT effects suggest the activation of inhibitory interneurons acting on a second order projecting cells to modulate afferent tactile and nociceptive information. PMID- 12763625 TI - Arsenite-induced formation of hydroxyl radical in the striatum of awake rats. AB - Recent studies on the mechanisms of arsenite toxicity report that some of its effects have been traced to the generation of reactive oxygen species during oxidative stress. In this study we analyze the formation of hydroxyl radicals in the brain of awake, freely moving rats, in order to obtain direct evidence of arsenic-induced oxidative stress in this tissue. We examined the time-course of hydroxyl radical formation in the striatum of both female and male rats who underwent a direct infusion during 60 min of different concentrations of arsenite in that structure through a microdialysis probe. We report here that basal levels of hydroxyl radical production in female rats are significantly higher than those in male rats (91.9+/-16.1 vs. 59.2+/-18.1 pmol/ml, P<0.001) and that the treatment with arsenite induced significant increases of hydroxyl radical formation over basal levels at 50, 100, 200 and 400 microM (95, 98, 98 and 99% increases, respectively, P<0.05 in all cases). The maximal response to 100 microM arsenite is significantly higher in female than in male rats (194.6+/-50.1 female rats and 88.1+/-11.6 pmol/ml male rats, P=0.036). These results support the participation of hydroxyl radicals in arsenic-induced disturbances in the central nervous system. PMID- 12763627 TI - Extracellular GABA concentrations in area 17 of cat visual cortex during topographic map reorganization following binocular central retinal lesioning. AB - Gamma-aminobutyric acid (GABA), the major inhibitory neurotransmitter in the central nervous system of mammals, plays an important role in cortical reorganization following sensory deprivation, by regulating the level of cortical inhibition and gating changes in receptive field size and synaptic efficacy. In cats it has been shown that 2 weeks after the induction of binocular retinal lesions, GABAergic inhibition, as determined by immunocytochemistry, is decreased in the deafferented region of area 17, whereas 3 months post-lesion, normal GABAergic control is restored within the cortical scotoma. In this study we used in vivo microdialysis to investigate the extracellular GABA concentrations 1-2 months post-lesion, in the sensory-deprived and remote, non-deprived region of area 17. Data were collected at those sample times and sites for which the extracellular glutamate concentrations had been determined in a previous investigation to elucidate the role of this excitatory neurotransmitter in cortical reorganization. As for glutamate, we observed significantly increased extracellular GABA concentrations in non-deprived area 17, whereas in deafferented area 17, extracellular GABA concentrations were comparable to those observed in normal, control subjects. These data suggest that 1-2 months post lesion the deafferented cortex behaves like normal visual cortex, in contrast to remote, non-deprived cortex. Notwithstanding the increase in extracellular GABA concentration of 134%, the parallel increase in glutamate concentration of 269% could give rise to a net increase in excitability in remote area 17. We therefore suggest that LTP-like mechanisms, and thereby cortical reorganization, might still be facilitated, while possible excessive hyperexcitability is balanced by the moderately increased GABAergic control. PMID- 12763626 TI - Effect of D-2 amino-5-phosphonopentanoate and nifedipine on postsynaptic calcium changes associated with long-term potentiation in hippocampal CA1 area. AB - The induction of long-term potentiation (LTP) in CA1 hippocampal area requires a rise in intracellular postsynaptic calcium. Two major calcium mechanisms may mediate the transmembrane calcium influxes that contribute to this calcium accumulation: the N-methyl-D-aspartate (NMDA) receptor channels, which are voltage dependent and have large calcium permeability and voltage-dependent calcium channels (VDCCs). We have addressed the relative contribution of these routes of calcium entry before and during LTP expression, in synaptically evoked dendritic calcium transients from a population of CA1 pyramidal neurons. Combining the use of the fluorescent calcium indicator Fura-2 with field potential measurements, we observed that the calcium transients evoked by single stimuli, during the maintenance phase of LTP, were enhanced. These transients were not affected by D-2 amino-5-phosphonopentanoate (D-APV) (50 microM), an antagonist of NMDA receptors but were reduced by approximately one-quarter, in the presence of the L-type VDCCs blocker nifedipine (10 microM). During tetanic stimulation (100 Hz, 1 s) the components triggered by the activation of those two calcium mechanisms had comparable magnitudes representing the sum about half of the intracellular calcium accumulation. Thus, following both single and high frequency stimulation, a substantial fraction of calcium entry may occur through other types of VDCCs or be due to calcium release from intracellular stores. PMID- 12763628 TI - Hypobaric hypoxia modifies constitutive nitric oxide synthase activity and protein nitration in the rat cerebellum. AB - Ischemic hypoxia provokes alterations in the production system of nitric oxide in the cerebellum. We hypothesize that the nitric oxide system may undergo modifications due to hypobaric hypoxia and that may play a role in high altitude pathophysiology. Therefore, changes in the nitric oxide system of the cerebellum of rats submitted to acute hypobaric hypoxia were investigated. Adult rats were exposed for 7 h to a simulated altitude of 8235 m (27000 ft.) and then killed after 0 h or 1, 3, 5 and 10 days of reoxygenation. Nitric oxide synthase calcium dependent and -independent activity, immunoblotting and immunohistochemistry of neuronal, endothelial, and inducible nitric oxide synthase, and nitrotyrosine were evaluated. Immunoreactivity for neuronal nitric oxide synthase slightly increased in the baskets of the Purkinje cell layer and in the granule cells, after 0 h of reoxygenation, although no changes in neuronal nitric oxide synthase immunoblotting densitometry were detected. Calcium-dependent activity significantly rose after 0 h of reoxygenation, reaching control levels in the following points, and being coincident with a peak of eNOS expression. Nitrotyrosine formation showed significant increments after 0 h and 1 day of reoxygenation. Nitrotyrosine immunoreactivity showed an intracellular location change in the neurons of the cerebellar nuclei and in addition, an appearance of nitration in the soma of the Purkinje cells was detected. No changes in inducible nitric oxide synthase activity, immunoblotting or immunohistochemistry were detected. We conclude that at least part of the nitric oxide system is involved in cerebellum responses to hypobaric hypoxia. PMID- 12763629 TI - Dynamic expression of p38beta MAPK in neurons and astrocytes after transient focal ischemia. AB - Here we report the dynamically regulated expression of p38beta MAPK isoform in specific subsets of cells in postischemic brain. The activity of p38beta MAPK in the postischemic brain revealed biphasic induction at 30 min and 4 days after 1 h MCAO. During the early surge period, p38beta MAPK was preferentially localized in the nucleus and dendrites of neurons in the future infarction area, while during the delayed surge p38beta MAPK was heavily induced in reactive astrocytes in penumbra. The temporally and spatially regulated pattern of p38beta MAPK expression in the postischemic brain suggests distinct roles of p38beta MAPK in neuronal death and in the astrocyte activation. PMID- 12763630 TI - Expression of metallothionein-III induced by hypoxia attenuates hypoxia-induced cell death in vitro. AB - Metallothioneins (MTs) are metal-binding proteins that are expressed in many tissues including brain. MTs protect cells and organs against metal toxicity and oxidants. Among MTs, a brain-predominant subtype MT-III has prominent neuroprotective activity against various types of damage. Here we show that the expression of MT-III is induced in cultured normal human astrocytes by hypoxia, and that overexpressed MT-III protects human embryonic kidney cells from hypoxia, suggesting that MT-III can protect the brain from hypoxic damage. PMID- 12763631 TI - Hypothalamic type II iodothyronine deiodinase: a light and electron microscopic study. AB - In the central nervous system, the active form of thyroid hormone, T3, derives from the cellular uptake and intracellular 5'-monodeiodination of T4 by type II 5'-monodeiodinase (DII). Here, we report that using an antiserum raised against the C-terminus of the full-length SeDII, immunolabeled cells were found in the rat hypothalamus in agreement with the DII mRNA distribution. Light and electron microscopy shows that DII is localized in astrocytes and tanycytes, supporting the hypothesis that these cells play an important role in the mediation peripheral signals, such as thyroid hormones, on hypothalamic functions. PMID- 12763632 TI - Glutamate induced currents in isolated inner hair cells from guinea-pig cochlea. AB - We investigated the direct action of glutamate (Glu) on the membrane current of isolated inner hair cells of guinea-pig cochlea. Glu elicited inward currents at a holding potential of -70 mV. Eight of 13 cells showed a steady inward current, while five of 13 cells showed a fast and rapidly desensitized current. I-V relationships demonstrated that the reversal potential of Glu-induced current was near 0 mV. Glu-induced currents were dose-dependent, where the half maximum concentration (K(d)) was 41 microM and Hill coefficient (n) was 1.75. PMID- 12763633 TI - Prolactin-releasing peptides. AB - Physiologic control of prolactin (PRL) secretion is largely dependent upon levels of dopamine accessing the adenohypophysis via the hypophysial portal vessels. However, it is clear that other factors of hypothalamic origin can modulate hormone secretion in the absence or presence of dopamine. Several neuropeptides have been identified as PRL releasing factors (PRFs) but none of these peptides appears to be a major determinant of PRL secretion in vivo. There remain uncharacterized activities in hypothalamic extracts that can alter secretion and production of the hormone. In addition, there exist a wide variety of substances (neurotransmitters, neuromodulators, neuropeptides) that can act within the hypothalamus to modify the neuroendocrine regulation of PRL secretion. These factors may not be considered true PRFs because their actions are not exerted directly at the level of the lactotroph; however, they can act in brain to stimulate PRL release in vivo and therefore might be considered PRL releasing peptides (PRPs). PMID- 12763634 TI - Receptor mechanisms of prenodal lymphatic constriction by dopamine. AB - It has been proposed that alterations in lymphatic smooth muscle activity significantly impact lymphatic function. Numerous endogenous vasoactive agents are known to constrict prenodal lymph vessels. In this study, we assessed the ability of dopamine to alter lymphatic smooth muscle tone in perfused prenodal lymph vessels. Additionally, the receptor mechanisms of dopamine's actions were elucidated. Both intralymphatic (i.l.) and intra-arterial (i.a.) dopamine significantly increased lymphatic perfusion pressure. The increase in lymphatic pressure was completely blocked by i.a. phentolamine, suggesting involvement of alpha(1)- and/or alpha(2)-adrenoreceptors. Intra-arterial infusion of the specific alpha(1)-receptor antagonist prazosin completely abolished the constriction seen during i.l. phenylephrine but only attenuated that produced by dopamine. Intralymphatic infusion of the DA(1)-receptor agonist SKF 82526-J and the DA(2)-receptor agonist LY 171555 caused significant relaxation of lymph vessels that had been previously constricted by i.a. norepinephrine infusion. These data indicate that the constriction produced by dopamine, in the concentrations employed in this study, is mediated by both alpha(1)- and alpha(2) adrenoreceptors. These lymph vessels do contain both DA(1)- and DA(2)-receptors but stimulation of these receptors results in lymphatic smooth muscle relaxation. PMID- 12763635 TI - Exogenous leptin inhibits the secretion of pancreatic juice via a duodenal CCK1 vagal-dependent mechanism in anaesthetized rats. AB - Leptin originally described as product of the ob gene has been shown to be expressed in various tissues including the gastrointestinal tract. In this study, we investigated the influence of leptin on the secretion of pancreatic juice in biliary-pancreatic duct cannulated anaesthetised rats and in dispersed rat pancreatic acini in vitro. Exogenous leptin was given in boluses intravenously with or without CCK-8 (12 pmol kg(-1) body weight) in the presence or absence pharmacological CCK(1) receptor blockade, cervical vagotomy, and capsaicin pre treatment. Administration of leptin (0.1, 1 and 10 microg kg(-1) body weight) did not affect the volume of bile and pancreatic juice while the protein and trypsin outputs were reduced in a dose-dependent manner. In the rats, leptin inhibited CCK-8 stimulated protein and trypsin outputs stronger than the basal pancreatic secretion. The inhibition by leptin was abolished by the pharmacological CCK(1) receptor blockade, cervical vagotomy, and capsaicin pre-treatment. In contrast, leptin did not affect basal and CCK-8-stimulated amylase release from the dispersed rat pancreatic acini in vitro. In conclusion, the results of the present study suggest that leptin does not act directly on the rat pancreatic acinar cells but inhibits the secretion of pancreatic enzymes acting indirectly via a neurohormonal CCK-vagal-dependent mechanism. PMID- 12763636 TI - Rat stomach ECL cells: mode of activation of histidine decarboxylase. AB - Histidine decarboxylase (HDC) occurs in ECL cells in the oxyntic mucosa of rat stomach. It is activated by gastrin. Refeeding of fasted rats or treatment with the proton pump inhibitor omeprazole promptly raised the serum gastrin concentration and consequently the HDC activity and the HDC protein content of the oxyntic mucosa. The food- and omeprazole-induced increase in HDC mRNA expression in the oxyntic mucosa was modest by comparison. Blockade of translation (cycloheximide) but not transcription (actinomycin D) prevented the postprandial rise in HDC activity. The half-life of HDC activity (after blockade of translation) was 94 min in omeprazole-treated rats and 55 min in fasted controls. The rate of enzyme synthesis was estimated to be 15 times higher in omeprazole-treated rats than in fasted controls. Inhibition of histamine uptake into ECL-cell granules by reserpine, a blocker of the vesicular monoamine transporter type-2, lowered the HDC activity and prevented the gastrin-induced HDC activation. We suggest that HDC activation reflects enhanced transcription, translation and/or posttranslational enzyme activation as well as stabilization, and that a high cytosolic histamine concentration suppresses HDC activation. PMID- 12763637 TI - Catestatin (CgA344-364) stimulates rat mast cell release of histamine in a manner comparable to mastoparan and other cationic charged neuropeptides. AB - Catestatin (bovine CgA(344-364)) is a cationic peptide, which besides reducing catecholamine secretion from chromaffin cells in vitro also acts a potent vasodilator in the rat in vivo. The alleged histamine releasing effect of catestatin was tested in vitro in rat mast cells. The most active domain of catestatin (bovine CgA(344-358): RSMRLSFRARGYGFR) caused concentration-dependent (0.01-5 microM) release of histamine from peritoneal and pleural mast cells. The potency and efficacy of catestatin was higher than for the wasp venom peptide, mastoparan. Only in the pleural cells was neurotensin (NT) more potent than catestatin, mastoparan and substance P (SP), consistent with a receptor-mediated histamine release by neurotensin. Amongst these cationic peptides, substance P was least effective. The acidic CgA peptide (WE-14, bovine CgA (324-337)) neither stimulated nor modulated histamine release by the cationic peptides. The catestatin and neurotensin evoked histamine release were suppressed by pertussis toxin (PTX), suggesting involvement of a G(i) subunit. Electron micrographs of rat pleural mast cells responding to catestatin revealed a concentration dependent discharge of granular material. We propose that catestatin activates histamine release from rat mast cells by a mechanism analogous to that already established for mastoparan and other amphiphilic cationic neuropeptides (the peptidergic pathway) and distinct from the mechanism of inhibition of catecholamine release from chromaffin cells. PMID- 12763638 TI - Molecular cloning of an unusual bicistronic cholecystokinin receptor mRNA expressed in chicken brain: a structural and functional expression study. AB - This report describes the molecular cloning and pharmacological characterization of a transiently expressed chicken brain cholecystokinin receptor (CCK-CHR) in COS-7 cells. A polymerase chain reaction (PCR)-based cloning strategy was applied using: (1) an initial PCR with deoxyinosine-containing primers designed to target conserved regions in CCK receptors, followed by (2) rapid amplification of cDNA ends (RACE), and (3) full-length PCR of the CCK-CHR cDNA. The full-length cloned bicistronic CCK-CHR cDNA contained a short upstream open reading frame (uORF) coding for a putative six-amino-acid-long peptide of unknown function, followed by a long open reading frame (lORF) encoding the 436-amino-acid-long CCK-CHR receptor protein. At the amino acid level, the CCK-CHR shared approximately 50% homology with mammalian and Xenopus laevis CCK receptors. The pharmacological profile of CCK-CHR resembled that of CCK-B receptors using agonists (CCK-8, CCK 4, gastrin-17), whereas CCK-CHR showed higher affinity for the CCK-A receptor antagonist, devazepide, than for the CCK-B receptor antagonist, L-365,260. To the best of our knowledge, this is the first description and functional expression study of a cloned chicken CCK receptor cDNA. PMID- 12763639 TI - Involvement of capsaicin-sensitive sensory nerves in cardioprotection of rutaecarpine in rats. AB - In the present study, we examined whether rutaecarpine protects against myocardial ischemia-reperfusion injury in rats and whether the protective effects of rutaecarpine are related to activation of capsaicin-sensitive sensory nerves. Rats were pretreated with rutaecarpine 10 min before the experiment, and then the left main coronary artery of rat hearts was subjected to 60-min occlusion followed by 3-h reperfusion. The infarct size, serum concentration of creatine kinase, and CGRP concentration in plasma were measured. Pretreatment with rutaecarpine (100 or 300 microg/kg, i.v.) significantly reduced infarct size and creatine kinase release concomitantly with a significant increase in plasma concentrations of CGRP. These effects of rutaecarpine were completely abolished by capsazepine (38 mg/kg, s.c.), a competitive vanilloid receptor antagonist, or by pretreatment with capsaicin (50 mg/kg, s.c.), which selectively depletes transmitters in capsaicin-sensitive sensory nerves. These results suggest that rutaecarpine protects against myocardial ischemia-reperfusion injury in rats and that the protective effects of rutaecarpine are related to activation of capsaicin-sensitive sensory nerves via activating vanilloid receptors. PMID- 12763640 TI - A multi-tyrosinated sst1/2 receptor preferring somatostatin agonist inhibits reflex and immune-mediated secretion in the guinea pig colon. AB - Somatostatin and its analogs such as WOC 3B were compared for their ability to alter the release of 5-hydroxytryptamine (5-HT) and prostaglandins and to affect chloride secretory capacity, determined by activity of neural reflexes or by the influence of immune mediators and other secretagogues. In guinea pig colon set up in flux chambers, the multi-tyrosinated sst1/sst2 receptor preferring somatostatin agonist, WOC 3B, inhibited stroking-evoked 5-HT release without affecting basal release. WOC 3B had no effect on stroking-induced or basal prostaglandin E2 release (PGE2). Neither 5-HT nor PGE2 release was dependent on neural input. Tetrodotoxin induced a decrease in basal short circuit current (Isc) indicative of a decrease in chloride secretion. The decrease in basal Isc during neural blockade was highly correlated with the decrease in basal Isc when WOC 3B was used. In piroxicam- and atropine-treated tissues, to eliminate prostaglandins and cholinergic muscarinic input to crypts, WOC 3B further reduced the piroxicam-resistant and not the atropine resistant Isc during brush stroking the mucosa. Somatostatin and WOC 3B reduced the stroking-evoked Isc with similar half maximum concentrations of 1-2 nM. WOC 3B reduced by more than 50% dimaprit evoked cyclical Isc. The rank order of potencies in inhibiting dimaprit-evoked Isc was: Somatostatin-14=WOC 3B>CH275=DC-32-92>DC-23-48>> >>DC-32-87=DC-32-97. Low nanomolar concentrations of WOC 3B primarily inhibited the neural effects of carbachol and forskolin on Isc without altering their epithelial effects. Equi molar concentrations (4 nM) of CH275, a somatostatin sst1 receptor agonist, and the somatostatin sst2 receptor agonist, [Tyr(3)]-octreotide, inhibited dimaprit evoked Isc by 25% and 26%, and their effects were additive. The results suggest that WOC 3B, a somatostatin analogue containing three tyrosine residues, has anti secretory effects due to activation of somatostatin sst1 and sst2 receptors on enteric neurons. PMID- 12763641 TI - Circulating adrenomedullin is increased in relation with increased creatinine and atrial natriuretic peptide in liver-transplant recipients. AB - OBJECTIVES: Circulating adrenomedullin (ADM), a potent vasorelaxing and natriuretic peptide involved in cardiovascular homeostasis, is increased after cardiac and renal transplantation. ADM is also implicated in hemodynamic abnormalities during liver cirrhosis, but whether ADM is increased late after liver transplantation is unknown. PATIENTS: A total of 18 subjects--10 liver transplant patients (Ltx) and 8 healthy subjects--were enrolled in the study. DESIGN AND MEASUREMENTS: After a 15-min rest period in supine position, heart rate and systemic blood pressure were determined in all subjects. Then, venous blood samples were obtained in order to simultaneously determine the cyclosporine through levels, the biological (cyclosporine, renal and hepatic functions) and hormonal (ADM and atrial natriuretic peptide (ANP)) characteristics of the Ltx. RESULTS: ADM (27.2+/-4.1 vs. 53.8+/-6.9 pmol/l, P=0.02), and ANP (5.9+/-0.9 vs. 12.8+/-1.4 pmol/l, P=0.001) were significantly increased in late, stable Ltx (35.4+/-9.6 months after transplantation). Furthermore, increased ADM correlated positively with elevated creatinine (r=0.76, P=0.01) and ANP (r=0.64, P=0.04) after liver transplantation. CONCLUSIONS: Liver-transplant patients exhibit a sustained increase in circulating ADM. Such an increase likely results from renal impairment associated with volume regulation abnormalities, suggesting a potential role for ADM in volume regulation after liver transplantation. PMID- 12763642 TI - [Three dimensional gait analysis and controlling spastic foot on stroke patients]. AB - INTRODUCTION: The purpose of this work is to assess the interest of three dimensional gait analysis in measuring the effect of orthesis and of selective tibial neurotomy without visual analysis. METHOD: After clinical examination, gait in the two groups was recorded by a three dimensional motion system (Vicon Oxford Metrics) with free velocity. The gait of ten stroke patients was compared bare-foot and with ankle-foot orthesis. The gait of nine stroke patients was compared bare-foot before and six month after selective tibial neurotomy. Kinematics data were studied in the sagittal plane, and muscular electrical activity was detected on the affected side by a ten-channel telemetry system using surface electrodes. An analogic visual scale ( EVA) was used by the patient, to assess discomfort during walking. RESULTS: In the "orthesis" group, calf spasticity was 2.5 on the Ashworth scale. With ankle-foot orthesis, the walking discomfort on EVA was significantly reduced from 4.25 to 1.16. Gait speed increased significativelly at 0.42 m s(-1) to 0.59 m s(-1). In swing phase, equinus disappeared. In stance phase equinus and recurvatum were controlled, anterior pelvic tilt decreased and hip extension increased. In the "neurotomy" group, 6 months after the tibial neurotomy, calf spasticity decreased from 3.4 to 0. The walking discomfort on EVA, was significantly reduced from 5.97 to 3.6. Gait speed increased from 0.51 m s(-1) to 0.54 m s(-1). Ankle dorsiflexion on the affected side, was possible during stance for five patients and residual motricity was released during swing phase for two patients. Stance knee recurvatum was corrected in five patients. CONCLUSION: Three dimensional analysis specifies the functional gait disability in stroke patients. It specifies the degree of correction provided by orthesis or tibial neurotomy in the various gait cycle phases and the impact on the superior joints. PMID- 12763643 TI - [3D preliminary analysis gait in stroke patients: knee flexion in compensatory strategies]. AB - OBJECTIVE: The aim of this preliminary study was to assess strategies of walking a stride in stroke patients with spastic right hemiplegia. MATERIAL AND METHODS: Gait was recorded in 3D in seven patients without other locomotion disorders. Kinematics data were studied only on the sagittal plane. The position and trajectory markers on the right side were studied during the swing phase in comparison with static standing position. Results were confronted with angular data. RESULTS: Three walking models were defined: 1) near normal gait with normal mobility in the knee; 2) gait with hicking while the flexion of the knee was reduced; 3) gait with a "talus" foot without motor recovery necessitating a pendular movement. DISCUSSION: The second pathological group was characterized with insufficient flexion in the knee in lifting the foot from the floor. In this group, patients adopted a compensation strategy with hicking in making the stride without touching the floor. We raised the question of limiting this adaptive strategy in order to enhance their remaining mobility. CONCLUSION: A 3D strategy gait analysis, before therapeutic choices, seems to confirm the value of analysing kinematic data in stroke patients with hemiplegia. The amplitude of knee mobility and hip compensation strategy can be specifically studied to improve the effectiveness of therapeutic strategies (orthesis, selective tibial neurotomy, botulinum toxin). PMID- 12763644 TI - [MRI study of the tissue volume progression of lower limbs in hemiplegic subjects]. AB - OBJECTIVES: The aim of this paper was to present the first results of a study about lower limbs' soft tissues (fat and muscle) evolution using magnetic resonance images. MATERIALS AND METHODS: Magnetic resonance images method (MRI) was chosen instead of X-ray scanner for its non-invasive and non-radiating properties. Three hemiplegics volunteers were followed during the first 6 months of their therapy. Lower limbs' MRI captures were made at the beginning and in the end of this period. Computerized processing of the data allows automatic recognition of the main lower limb's tissues. Volumes were determined from images and classified in 4 categories (i.e. muscle, fat, spongy bone, cortical bone). RESULTS: Data analysing showed a different evolution of soft tissues in healthy lower limb compared to pathological lower limb. DISCUSSION-CONCLUSION: This results should lead us to a better understanding of therapeutic efficiency. PMID- 12763645 TI - [Functional surgery of upper limb in tetraplegics since 50 years]. AB - OBJECTIVE: Development of upper limb functional surgery in tetraplegics in the last 50 years. METHODS: The literature review relating to the years 1950-2002 was carried out with 3 data bases: Medline, Pascal, Embase. This review also involved a thorough study of non-indexed references. RESULTS: A large number of surgical procedures are described. Two priorities are stressed by the authors: safety of these procedures and duration of postoperative immobilization. CONCLUSION: This review of literature shows that the prospects for restoring upper limb function in tetraplegics are greater than ever, offering a larger number of patients the possibility to increase their independence in daily life. Functional surgery remains, nevertheless, demanding in terms of length of immobilization and presupposes requiring a multidisciplinary approach requiring rehabilitation teams to be up to date with surgical procedures. PMID- 12763646 TI - [Functional surgery of upper limb in tetraplegics]. PMID- 12763647 TI - [Whiplash injury of the neck from concepts to facts]. AB - OBJECTIVES: To focus on a topic of traumatology and rehabilitation becoming recently a much debated public health problem. METHOD: A references search from Medline database with whiplash as keyword was carried out. Were selected articles with abstracts in french or english and focusing on accidentology, biomechanics, demonstrated lesions, epidemiology and treatments. RESULTS: From 1664 references found, 232 were reviewed. The usual mechanism of crash is a rear-end collision inducing in the occupants of the bumped vehicle a sudden lower cervical spine extension with upper flexion followed by a global flexion. In nearly 50% of the cases, the stress occurring in the collision is comparable to that observed in bumper cars. The velocity changes are seldom up to 15 km/h. A headrest at the level of the center of gravity of the head restrict significantly the extension of the neck. Every structure of the cervical spine could be damaged and mainly the facet joints but the lesions were only demonstrated in severes traumatisms. The discrepancies in incidence among the different countries could be related to their medicolegal system. Although subjectives, the early symptoms are rather similar among patients suggesting true anatomical or functional disorders but the chronicity seems to be mainly related to social and psychological factors. The association of: no posterior midline cervical tenderness, no intoxication, normal alertness, no focal neurological deficit and no painful distracting injuries has a good predictive value of the lack of osteo-articular lesion on X-rays. Except the grade IV of the Quebec task Force (0, no symptom; 1, pain and stiffness; 2, neck complaint and physical signs; 3, neck complaint and neurological signs; 4, fracture or dislocation) the use of a collar should be avoided and the cervical spine should be mobilized. CONCLUSION: In most whiplash injuries, the mildness should be early stated, mobilization encouraged, and procedures of compensation shortened. PMID- 12763650 TI - Chemical speciation of insulinomimetic VO(IV) complexes of pyridine-N-oxide derivatives: binary and ternary systems. AB - In order to estimate the impact of the low-molecular-mass (l.m.m.) VO(IV) binders of blood serum on the potentially insulin-enhancing compound VO(HPO)(2) (HPO, 2 hydroxypyridine-N-oxide): and VO(MPO)(2) (MPO, 2-mercaptopyridine-N-oxide), the speciation in the binary system VO(IV)-HPO and VO(IV)-MPO and in the ternary systems VO(IV)-HPO(MPO)-ligand B (B=oxalate, lactate, citrate or phosphate) was studied by pH-potentiometry. The stability constants of the complexes formed were determined in aqueous solution at I=0.2 M (KCl) and T=25 degrees C. The most probable binding modes of the complexes were determined by EPR method. The pyridine-N-oxides were found to form very stable bis complexes, which are predominant in the pH range 2-7. The results in the ternary systems demonstrate that only the citrate is a strong enough VO(IV) binder to compete with the carrier ligands. The binding ability of the high-molecular-mass (h.m.m.) serum proteins albumin and transferrin were also assessed and transferrin was found to be an efficient binder molecule. The actual solution state of these compounds in blood serum is compared with that of other insulin-mimic VO(IV) complexes. PMID- 12763651 TI - Synthesis, structure and nuclease properties of several ternary copper(II) peptide complexes with 1,10-phenanthroline. AB - Three new ternary peptide-Cu(II)-1,10-phenanthroline (phen) complexes, [Cu(L-ala gly)(phen)].3.5H(2)O 1, [Cu(L-val-gly)(phen)] 2 and [Cu(gly-L-trp)(phen)].2H(2)O 3, have been prepared and structurally characterised. These compounds exist as distorted square pyramidal complexes with the five co-ordination sites occupied by the tridentate peptide dianion and the two heterocyclic nitrogens of the phenanthroline ligand. The bulk of the lateral chain in the peptide moiety determines the relative disposition of the phen ligand. Thus, in [Cu(L-val gly)(phen)] 2, the phenanthroline plane is deviated towards the opposite side of the isopropyl group of the L-valine moiety. On the other hand, in [Cu(gly-L trp)(phen)].2H(2)O 3 the absence of stacking interactions between phen and indole rings and the presence of an intramolecular CH...pi interaction should be pointed out. These complexes exhibit significant differences in their nuclease activity which depends on the nature of the peptidic moiety, the complex [Cu(gly-L-trp) (phen)].2H(2)O 3 being the most active. PMID- 12763653 TI - Isomer abundance of bis(beta-diketonato) complexes of titanium(IV). Crystal structures of the antitumor compound budotitane [Ti(IV)(bzac)(2)(OEt)(2)] and of its dichloro-derivative [Ti(IV)(bzac)(2)Cl(2)] (bzac=1-phenylbutane-1,3-dionate). AB - The molecular tumor inhibiting titanium compound budotitane [Ti(IV)(bzac)(2)(OEt)(2)] (1) and its dichloro-derivative [Ti(IV)(bzac)(2)Cl(2)] (2) (bzac=1-phenylbutane-1,3-dionate) have been crystallized and characterized by X-ray crystallography and further physical methods. Budotitane (1) crystallizes in the tetragonal, non-centrosymmetric space group P4(1) with two molecules in the asymmetric unit. Both molecules adopt the cis-cis-trans configuration with the acetyl ends of the benzoylacetonate ligands in the trans position. The dichloro-derivative of budotitane, [Ti(IV)(bzac)(2)Cl(2)] (2) crystallizes in the monoclinic, centrosymmetric space group P2(1)/n with one molecule only in the asymmetric unit. In contrast to budotitane (1), (2) shows a cis-trans-cis arrangement with the benzoyl groups in the trans position. In both complexes there are equal numbers of Delta and Lambda enantiomers within the unit cell. The phenyl groups in (1) as well as in (2) are in approximately coplanar conjugation to the metal enolate rings. The thermal degradation of budotitane (1) was investigated in the temperature range from 25 degrees C up to 800 degrees C and reveals the formation of Ti(IV)O(bzac(2-)) as an intermediate and of the rutile phase of TiO(2) as a final product. It may be worthwhile to introduce budotitane in the form of isomerically pure crystals in the preparation of the drug used for future tests. PMID- 12763652 TI - Stoichiometric preference in copper-promoted oxidative DNA damage by ochratoxin A. AB - The ability of the fungal carcinogen, ochratoxin A (OTA, 1), to facilitate copper promoted oxidative DNA damage has been assessed using supercoiled plasmid DNA (Form I)-agarose gel electrophoresis and gas chromatography-mass spectrometry with selected-ion monitoring (GC-MS-SIM). OTA is shown to promote oxidative cleavage of Form I DNA with optimal cleavage efficiency occurring under excess Cu(II) conditions. As the concentration of OTA was increased and present in excess of Cu(II) the cleavage was less effective. Parallel findings were found for the ability of the OTA-Cu mixture to facilitate oxidative base damage. Yields (lesions per 10(6) DNA bases) of modified bases upon exposure of calf-thymus DNA (CT-DNA) to OTA-H(2)O(2)-Cu(II) were diminished when the OTA:Cu ratio was increased to 5:1. Electrochemical studies carried out in methanol implicate a ligand-centered 2e oxidation of OTA in the presence of excess Cu(II), while product analyses utilizing electrospray mass spectrometry support the intermediacy of the quinone, OTQ (3), in Cu-promoted oxidation of OTA. The implications of these findings with regard to the mutagenicity of OTA are discussed. PMID- 12763654 TI - Pt(II) complexes with N-(3-pyridyl)-2-(4 (trifluoromethyl)phenyl)diazenecarboxamide and their reactions with glutathione. AB - The reaction between [PtCl(dmso)(en)]Cl (dmso=dimethyl sulfoxide, en=ethylenediamine) and N-(3-pyridyl)-2-(4 (trifluoromethyl)phenyl)diazenecarboxamide (L) was studied using multinuclear NMR spectroscopy. The water-soluble complexes [PtCl(en)(L-N1)](+) (1) and [Pt(en)(L N1)(2)](2+) (2) were isolated and their reactions with glutathione (GSH) were investigated to assess the oxidation properties of coordinated L. Both species 1 and 2 oxidized GSH to GSSG, while the reduced form of L (semicarbazide, SL) remained coordinated to Pt(2+). In complex 1 the labile chloride ion was substituted by the thiol moiety of GSH, which gave rise to the release of en in excess GSH over a period of 7 days. Complexes [PtCl(dmso)(en)]Cl, 1, 2 and ligand L were tested against T24 bladder carcinoma cells. Ligand L and complexes 1 and 2 showed higher cytotoxicity than [PtCl(dmso)(en)]Cl. PMID- 12763655 TI - Ca(2+) and Na(+) binding to high affinity sites of calcium-containing proteins measured by capillary electrophoresis. AB - A method for the determination of stability constants of metal ion-protein binding, based on capillary electrophoresis, is presented. It utilizes the change in electrophoretic mobility of the protein upon binding of a metal ion. Taking advantage of edta(4-) as a controller of the free Ca(2+) concentration, a [Ca(2+)](free) as low as 10(-9) M has been attained in the solutions. We have found this method very useful for measuring binding of Ca(2+) to proteins, where the stability constant is in the range 10(5)-10(8) M(-1). The stability constants for the binding of Ca(2+) to proteinase K and bovine alpha-lactalbumin has by this method been measured at an ionic strength of 0.1 M, pH(c) 7.40 and 25 degrees C. For proteinase K a constant of 10(7.4) M(-1) is found, and for alpha lactalbumin the constant has been found to be 10(9.2) M(-1). The structural stability of both proteins are found to be affected by the presence of Na(+) in the buffer solutions. From this observation, association constants for binding of Na(+) to the Ca(2+) sites have been calculated to 10(2.4) M(-1) for proteinase K and 10(3.5) M(-1) for alpha-lactalbumin. Less than 50 microg have been used of each protein in this study, an obvious advantage over other methods. PMID- 12763656 TI - Studies on the interaction between Ag(+) and human serum albumin. AB - The interaction between Ag(+) and human serum albumin (HSA) has been intensively studied by means of equilibrium dialysis, ligand-to-metal charge transition (LMCT) bands, circular dichroism (CD) and Raman spectroscopy. Scatchard analysis of the results of equilibrium dialysis indicates the presence of two types of binding sites for Ag(+) on HSA, and the orders of magnitude of binding stability constants are found to be 10(5) and 10(4), respectively. During the binding process, a gradual increase in absorbance values of LMCT bands is observed with time-scanning UV absorption spectra, implying the Ag(I) centers are continually formed in HSA. The time-scanning CD spectra provide evidence that the binding of Ag(+) induces HSA to undergo a slow rearrangement of tertiary structure, and to change from the original conformation in the absence of Ag(+) (B-state) to conformation binding with Ag(+) (A-state). The rate constants and activation free energy of A-B transition are calculated. The Raman spectrum of Ag(I)-HSA system shows distinct vibration bands at 224 and 246 cm(-1) in the low-frequency region, which significantly reveal the formation of Ag-S and Ag-N bonds. In addition, the electrostatic interaction between Ag(+) and negatively charged oxygen is also detected with Raman spectroscopy. PMID- 12763657 TI - Speciation study of the anti-inflammatory drug tenoxicam (Htenox) with Cu(II): X ray crystal structure of [Cu(tenox)(2)(py)(2)].EtOH. AB - A speciation study was carried out in aqueous solution of the anti-inflammatory drug tenoxicam (Htenox), under quasi-physiological conditions (temperature of 37 degrees C and ionic strength 0.15 M NaCl) in order to determine the acidity constants from spectrophotometric studies, the pK(a) values found being pK(1)=1.143+/-0.008 and pK(2)=4.970+/-0.004. Subsequently, the spectrophotometrical speciation of the different complexes of Cu(II) with the drug was performed under the same conditions of temperature and ionic strength, observing the formation of Cu(Htenox)(2)(2+) with log beta(212)=20.05+/-0.01, Cu(tenox)(2) with log beta(012)=13.6+/-0.1, Cu(Htenox)(2+) with log beta(111)=10.52+/-0.08, as well as Cu(tenox)(+) with log beta(011)=7.0+/-0.2, all of them in solution, and solid species Cu(tenox)(2)(s) with an estimated value of log beta(012)(s) approximately 18.7. The crystalline structure of the complex [Cu(tenox)(2)(py)(2)]. EtOH, was also determined, and it was observed that tenoxicam employs the oxygen of the amide group and the pyridyl nitrogen to bond to the cation. PMID- 12763658 TI - Effect of cadmium and zinc ethanolamine complexes on rat brain monoamine oxidase B activity in vitro. AB - Monoamine oxidase-B (MAO-B) from rat brain was inhibited strongly by the prepared cadmium and zinc ethanolamine complexes obtained from their sulphate and chloride salts. The inhibition of MAO-B by these complexes was time-dependent and fully reversible after dilution and sedimentation. In vitro, the cadmium ethanolamine complexes were more potent at inhibiting MAO-B than the zinc complexes. The inhibitory effect of these complexes follow the order: TEA>DEA>MEA, due to the alkyl residues and steric effect properties. The inhibition of MAO-B by cadmium and zinc ethanolamine complexes was a noncompetitive type. The K(i) values were calculated. The influence of the complexes on the activity of MAO-B was rather evaluated. It decreased the MAO-B activity. The IC(50) values of the two potent cadmium and zinc triethanolamine complexes on MAO-B were evaluated indicating that the complexes were tightly binding, but reversible inhibitors for MAO-B. In general, these systems may be used for preventing some neurodegenerative diseases. PMID- 12763659 TI - The preparation, characterisation and in vitro cytotoxicity of potentially chemotherapeutic heterobimetallic complexes containing early and late transition metals. AB - The reactions of phosphine Ph(2)P(CH(2))(2)SO(3)Na with Cp(2)M'Cl(2) (M'=Ti, Zr) in aqueous solution give the metallophosphines, Cp(2)Ti(OSO(2)(CH(2))(2)PPh(2))(2) (Cp=cyclopentadienyl) and CpZr(OH)(OSO(2)(CH(2))(2)PPh(2))(2). These react with CODM"Cl(2) (M"=Pd, Pt) (COD=1,5-cyclooctadiene) in dichloromethane to give heterobimetallic complexes Cp(2)Ti(OSO(2)(CH(2))(2)PPh(2))(2)M"Cl(2) and CpZr(OH)(OSO(2)(CH(2))(2) PPh(2))(2)M"Cl(2) respectively. The compounds are characterised by infrared and NMR spectroscopies and elemental analysis. Electrospray mass spectra of the complexes are reported and compared to those of Cp(2)M'Cl(2) in water and dimethylsulfoxide (DMSO). For zirconocene dichloride and its product heterobimetallic complexes, the addition of ethylenediamine tetraacetic acid disodium salt (Na(2)H(2)EDTA) was found to be an effective ionisation enhancement agent for the electrospray mass spectral studies. Cytotoxicity studies for the previously reported Cl(2)Pt(PPh(2)(CH(2))(2)SO(3)H)(2).3.5H(2)O (Wedgwood et al., Inorg. Chim. Acta 290 (1999) 189), and the compounds Cp(2)Ti(OSO(2)(CH(2))(2) PPh(2))(2).1.5H(2)O and Cp(2)Ti(OSO(2)(CH(2))(2)PPh(2))(2)PtCl(2).4H(2)O reported here, have been evaluated by colony formation assay against cisplatin-sensitive and -resistant cell lines L929 and L929/R to highlight potential chemotherapeutic activity. The compound Cl(2)Pt(PPh(2)(CH(2))(2)SO(3)H)(2).3.5H(2)O overcomes cisplatin resistance. PMID- 12763660 TI - The intracellular iron sensor calcein is catalytically oxidatively degraded by iron(II) in a hydrogen peroxide-dependent reaction. AB - The fluorescent metal chelating dye calcein is used to obtain an estimate of cellular iron levels and to measure the kinetics of the entry of chelators and chelating drugs into cells. Under reducing conditions in the presence of ascorbic acid, such as that would be present in the cell, the Fe(II)-calcein complex was rapidly formed with a rate constant of 3 x 10(5) M(-1) s(-1). A slower iron dependent catalytic degradation of calcein also occurred that resulted in the formation of a non-fluorescent calcein product. The Fe(II)-catalyzed degradation of calcein was largely, but not completely, prevented by catalase. Electron paramagnetic resonance spin trapping experiments showed that the Fe(II)-calcein complex promoted formation of hydroxyl or a hydroxyl radical-like species. Together these results indicated that Fe(II) catalyzed the degradation of calcein through both hydrogen peroxide, and to a lesser extent, non-hydrogen peroxide dependent pathways. The iron-calcein complexes that were responsible for the degradation of calcein were likely high valence oxidizing iron-oxo species such as perferryl or ferryl complexes that were redox cycled by ascorbic acid. Thus, the use of calcein as an intracellular iron-sensing indicator may yield misleading results due to its degradation under certain conditions. PMID- 12763661 TI - The interaction of metal ions and Marimastat with matrix metalloproteinase 9. AB - The effect of a range of metal ions on the ability of Marimastat to inhibit matrix metalloproteinase 9 (MMP-9) was examined in a fluorescence based proteolytic assay. Whilst none of the metals examined significantly affected the inhibitory ability of Marimastat, several metal ions did have a significant effect on MMP-9 activity itself. In the absence of Marimastat, Zn(II) and Fe(II) significantly inhibited MMP-9 activity at metal ion concentrations of 10 and 100 microM, respectively. In both the absence and presence of Marimastat, Cd(II) significantly inhibited MMP-9 at 100 microM. In contrast, 1 mM Co(II) significantly upregulated MMP-9 proteolytic activity. PMID- 12763662 TI - Oxidation of 2,4-dichlorophenol catalyzed by horseradish peroxidase: characterization of the reaction mechanism by UV-visible spectroscopy and mass spectrometry. AB - The hydrogen peroxide-oxidation of 2,4-dichlorophenol catalyzed by horseradish peroxidase has been studied by means of UV-visible spectroscopy and mass spectrometry in order to clarify the reaction mechanism. The dimerization of 2,4 dichlorophenol to 2,4-dichloro-6-(2,4-dichlorophenoxy)-phenol and its subsequent oxidation to 2-chloro-6-(2,4-dichlorophenoxy)-1,4-benzoquinone together with chloride release were observed. The reaction rate was found to be pH-dependent and to be influenced by the pK(a) value of 2,4-dichlorophenol. The dissociation constants of the 2,4-dichlorophenol/horseradish peroxidase (HRP) adduct at pH 5.5 and 8.5 were also determined: their values indicate the unusual stability of the adduct at pH 5.5 with respect to several adducts of HRP with substituted phenols. PMID- 12763663 TI - Studies on the interaction of altromycin B and its platinum(II) and palladium(II) metal complexes with calf thymus DNA and nucleotides. AB - The interaction of the anticancer antibiotic altromycin B and its isostructrural Pt(II) and Pd(II) metal complexes with native calf thymus (CT) DNA was studied using UV-thermal denaturation experiments, circular dichroism spectroscopy and temperature controlled spectrophotometric titrations. Altromycin B stabilizes the double helix by raising the T(m), mainly by intercalation of its chromophore between the base pairs and interacting electrostatically via its sugar moieties with the edges of the DNA helix. Moreover, altromycin B induces a B-->A structural transition of CT DNA. The effect on DNA stability and conformation depends on the metal ion. Pt(II) and Pd(II) complexes induce the B-->A structural transition and stabilize the double helix similarly but they present lower final hyperchromicity due to premelting effects which were caused by intra- and interstrand crosslinking. Thus, a synergic effect of the metal ions to altromycin B-CT DNA interaction is observed in both cases. Altromycin B interacts with 5' GMP, 5'-AMP and 5'-CMP by electrophilic attack of the opened epoxide ring to the N(7)G, N(1)/N(7)A and N(3)C. Thus, covalent binding between these nucleotides and altromycin B takes place and explain the multiple binding mode suggested by the studies of the interaction of altromycin B and its complexes with DNA. The [Pd(II)-altroB] complex dissociates in the presence of the nucleotides, and various species of Pd(II)-nucleotide complexes, especially with 5'-GMP, are formed. The [Pt(II)-altroB] complex dissociates too, but only one or two species of Pt(II)-nucleotide complexes are formed, and in the case of 5'-AMP interaction the formation of a tertiary altroB-Pt(II)-5'AMP complex is proposed. 5'-TMP reacts very weakly in comparison with the other three nucleotides. These interactions were followed by 1H-NMR. PMID- 12763664 TI - A functionalized cobalt(III) mixed-polypyridyl complex as a newly designed DNA molecular light switch. AB - The ligand ODHIP (3,4-dihydroxyl-imidazo[4,5-f][1,10]phenanthroline) and its cobalt(III) complex [Co(bpy)(2)(ODHIP)](3+) were synthesized and characterized. Binding of this complex with calf thymus DNA has been investigated by spectroscopic methods and viscosity. The experimental results indicated that the complex bound to DNA by intercalation. In Tris buffer, the complex could emit relatively weak luminescence. After binding to DNA, the notable enhancement was observed. However, when the Cu(2+) was further added, the luminescence decreased gradually and disappeared after the equimolar concentrations of Cu(2+) was added, which exhibited the "off-on-off" properties of molecular light switch. PMID- 12763665 TI - Interactions of oxovanadium(IV) and the quinolone family member--ciprofloxacin. AB - The interactions of quinolone ciprofloxacin (cfH) and oxovanadium(IV) were studied by various methods. Green crystals of a complex [V(IV)O(cf)(2)(H(2)O)] were isolated and the molecular connectivities established, although the crystal structure was not perfectly refined due to the instability of the crystals. Based on a plausible interpretation of the data sets, two cf anions bidentately coordinate to a vanadyl cation through carboxylate and carbonyl oxygen atoms; in addition, there is a water molecule in the coordination sphere. Solution techniques (cyclic voltammetry, electronic and electron paramagnetic resonance spectroscopy, potentiometric measurements) confirmed the presence of various species in the solution, the composition of which strongly depends on the conditions in the system. The antibacterial activity of the complex against various microorganisms was tested and it was established that its activity is similar to that of free ciprofloxacin. PMID- 12763666 TI - Ligand-exchangeability of 2-coordinate phosphinegold(I) complexes with AuSP and AuNP cores showing selective antimicrobial activities against Gram-positive bacteria. Crystal structures of [Au(2-Hmpa)(PPh(3))] and [Au(6-Hmna)(PPh(3))] (2 H(2)mpa=2-mercaptopropionic acid, 6-H(2)mna=6-mercaptonicotinic acid). AB - Selective and effective antimicrobial activities against Gram-positive bacteria (B. subtilis and/or S. aureus) were found in 2-coordinate gold(I)-PPh(3) complexes with AuSP and AuNP cores, i.e. [Au(L)(PPh(3))] (HL=2-H(2)mna [H(2)mna=mercaptonicotinic acid] 3, D-H(2)pen [H(2)pen=penicillamine] 4, D,L H(2)pen 5, 4-H(2)mba [H(2)mba=mercaptobenzoic acid] 8, Hpz [Hpz=pyrazole] 9, Him [Him=imidazole] 10, 1,2,3-Htriz [Htriz=triazole] 11, 1,2,4-Htriz 12, Htetz [Htetz=tetrazole] 13), whereas no activity was observed in 2-coordinate AuSP core complexes [Au(2-Hmba)(PPh(3))] 6 and [Au(3-Hmba)(PPh(3))] 7. The two novel AuSP core complexes, [Au(2-Hmpa)(PPh(3))] [H(2)mpa=mercaptopropionic acid] 1 and [Au(6 Hmna)(PPh(3))] 2, were prepared and characterized by elemental analysis, FT-IR, TG/DTA, and ((31)P, 1H and 13C) NMR spectroscopy. The crystal structures of 1 and 2 were determined as a supramolecular arrangement of the 2-coordinate AuSP core. Both 1 and 2 significantly showed antibacterial activities. As a model reaction of phosphinegold (I) complexes with the cysteine residue in the biological ligands, we examined if the ligand exchange reactions of the aromatic anions L(1)(-) in [Au(L(1))(PPh(3))] (HL(1)=6-H(2)mna 2, 2-H(2)mna 3, 2-H(2)mba 6, Hpz 9, Him 10, 1,2,3-Htriz 11, 1,2,4-Htriz 12) with aliphatic thiols HL(2) (HL(2)=2 H(2)mpa, D-H(2)pen) occurred under the mild conditions and, also, if the 'reverse' reactions, namely, the ligand exchange reactions of the thiolate anions in [Au(2-Hmpa)(PPh(3))] 1, [Au(D-Hpen)(PPh(3))] 4 and [Au(2-Hmba)(PPh(3))] 6 with the free ligands HL(1) took place under similar conditions. In this work, a relationship of the ligand-exchangeability among 2-coordinate gold(I) complexes (1-4, 6, 9-12) was revealed. Complex 6 was substitution-inert, whereas complexes 1-4 and 9-12 were substitution-labile. The ligand-exchangeability of Au-S and Au N bonds in the 2-coordinate phosphinegold(I) complexes with AuSP and AuNP cores to form new AuSP cores, with retention of the Au-P bond, was closely related to the observed activities against Gram-positive bacteria, and the ease of the ligand-exchange reaction was strongly related to the intensity of the activities. PMID- 12763667 TI - Metal chelates of 2-hydroxy-4-methylthiobutanoic acid in animal feeding: preliminary investigations on stability and bioavailability. AB - The alpha-hydroxyacid 2-hydroxy-4-methylthiobutanoic acid (the so-called methionine hydroxy-analogue, MHA), largely used in animal nutrition as a source of methionine, forms stable metal chelates with divalent metals of formula [[CH(3)SCH(2)CH(2)CH(OH)COO](2)M].ZnH(2)O. Protonation and zinc(II) complex formation constants have been determined by pH-metry at 25 degrees C; the ternary system Zn(2+)/MHA/glycine was also studied by pH-metry and the formation constant of the species [ZnLA] was determined [log beta=6.57(11)]. Experiments in vitro with human intestinal CACO-2 cells indicated that the MHA/Fe chelate was taken up by the cells without any apparent toxic effect. PMID- 12763668 TI - The structure of the Ce(III)-angiotensin II complex as obtained from NMR data and molecular dynamics calculations. AB - Angiotensin II is shown by nuclear magnetic resonance (NMR) to form a complex in water at pH 4.0 with cerium(III), the ideal paramagnetic probe for Ca(2+). Paramagnetic shifts induced by the metal were used for the determination of dissociation constant and complex stoichiometry. ROESY cross-peaks and 3 J(HN H)(alpha) coupling constants were converted into distance and angular constraints to determine the structure of the complex by molecular dynamics using the simulated annealing protocol. The complex is kinetically labile and involves the Asp-1 side chain and the Phe-8 terminal carboxylates as binding groups resembling a hairpin which has been suggested as a possible biologically active structure. PMID- 12763669 TI - Effects of three PBDEs on development, reproduction and population growth rate of the harpacticoid copepod Nitocra spinipes. AB - The current knowledge concerning effects of polybrominated diphenyl ethers (PBDEs) on aquatic organisms is very limited. A full life-cycle (< or =26 days exposure) ecotoxicity test with the particle-feeding copepod Nitocra spinipes was therefore used to study effects of BDE-47, -99 and -100 on larval development rate (LDR) and population growth rate (r(m)). LDR significantly decreased in copepods exposed for 6 days to nominal concentrations > or =0.013 mg/l BDE-47 and > or =0.03 mg/l BDE-99. Large concentration ratios (< or =338) between adult acute and juvenile subchronic endpoints were observed. Exposure over the full life cycle (< or =26 days) showed that r(m) in general was a less sensitive endpoint than LDR. Still, the r(m) in copepods exposed to 0.04 mg/l BDE-47 was significantly reduced compared to the controls (***P<0.001). Partitioning experiments with 14C-BDE-47 and 14C-BDE-99 in the test system showed that the major fractions (approximately 50-80%) were associated to particulate material. Our findings indicate that development and reproduction in N. spinipes are sensitive to the tested PBDEs and that ingestion of particle-adsorbed PBDEs most likely is the predominant route of exposure in N. spinipes. However, to further improve the usefulness of laboratory effect levels of PBDEs and other lipophilic substances for environmental risk assessment, it is important to develop ecotoxicological tools, which can evaluate and rate the toxic contribution from different matrices, such as suspended particles, sediment, food, water etc. PMID- 12763670 TI - Developmental toxicity of methoprene and several degradation products in Xenopus laevis. AB - Methoprene is an insect juvenile growth hormone mimic, which inhibits pupation and is used for the control of emergent insect pests such as mosquitoes. Researchers have hypothesized that methoprene use in US may be a contributing factor to the recent increase in malformed amphibians. However, little is known concerning the developmental toxicity of methoprene and its degradation products in amphibians. In these studies, the aqueous stability and developmental toxicity of methoprene and several degradation products (methoprene acid, methoprene epoxide, 7-methoxycitronellal, and 7-methoxycitronellic acid) were examined. Xenopus laevis embryos (stage 8) were exposed to the test chemicals for 96 h. Assays were conducted under static renewal (24 h) conditions and chemical concentrations in water were measured at the beginning and end of the renewal periods. Methoprene exposure did not result in developmental toxicity at concentrations up to 2 mg/l, which is slightly higher than its water solubility. Methoprene acid, a relatively minor degradation product, produced developmental toxicity when concentrations exceeded 1.25 mg/l. Methoprene epoxide and 7 methoxycitronellal caused developmental toxicity at concentrations of 2.5 mg/l and higher. 7-Methoxycitronellic acid was not developmentally toxic at a test concentration as high as 30 mg/l. The five test chemicals had differential stability in aqueous solution that was in some instances affected by the presence of test organisms. These data indicate that methoprene and its degradation products are not potent development toxicants in X. laevis. This, in combination with the fact that field applications of sustained-release formulations of methoprene result in methoprene concentrations that do not typically exceed 0.01 mg/l, suggests that concerns for methoprene-mediated developmental toxicity to amphibians may be unwarranted. PMID- 12763671 TI - Toxic vents and DNA damage: first evidence from a naturally contaminated deep-sea environment. AB - Levels of DNA strand breakage were measured, using the comet assay, in cells from vent mussels, Bathymodiolus azoricus, from three contrasting vent fields on the mid Atlantic Ridge. Different levels of DNA damage were recorded in untreated mussels, shortly after collection, and it was animals from the shallowest, and less active, Menez Gwen vent field (840-m depth), which showed the greatest amount of damage. In contrast to animals from two deeper and putatively more toxic sites, Menez Gwen animals went on to repair this damage and were able to survive under laboratory conditions at 1 bar pressure for several months. Animals from the two deeper sites showed both higher levels of initial mortality and a much reduced capacity for survival at 1 bar. The differences in DNA damage levels at the time of collection were interpreted as an expression of differences in cell viability/enzyme activity rather than a reflection of any differences in their natural environmental conditions. Small B. azoricus showed a capacity to repair DNA damage, whereas this ability appeared to be lacking in large individuals. By reproducing at a relatively early age, the deep-sea vent fauna may be able to resist the toxic effects of its environment by exploiting this natural, stage specific capacity to repair damaged DNA. PMID- 12763672 TI - Application of the micronucleus and comet assays to mussel Dreissena polymorpha haemocytes for genotoxicity monitoring of freshwater environments. AB - Assessment of DNA damage is of primary concern when determining the pollution related stress in living organisms. To monitor genotoxicity of the freshwater environments we used micronucleus (MN) and comet assay on Dreissena polymorpha haemocytes. Caged mussels, collected from the river Drava, were transplanted to four monitoring sites of different pollution intensity in the river Sava. Exposition lasted for a month. The baseline level of MN frequencies in the haemocytes of mussels from reference site (river Drava) was 0.5 per thousand. No increase in MN frequency was found in mussels from the medium-polluted site (Zagreb) in the river Sava while other, more polluted sites showed higher MN frequencies ranging from 2.7 per thousand (Lukavec) and 3.1 per thousand (Oborovo) to 5.2 per thousand (Sisak). Results from comet assay showed concordance with MN assay in indicating intensity of DNA damage. The use of haemocytes from caged, non-indigenous mussels in MN and comet assay proved to be a sensitive tool for the freshwater genotoxicity monitoring. PMID- 12763673 TI - Effects and toxicity of phthalate esters to hemocytes of giant freshwater prawn, Macrobrachium rosenbergii. AB - Phthalate esters (PAEs) have been considered as environmental pollutants and have been subject to control in the United States of America and Japan. The aim of this study was to investigate the effects and toxicity of eight PAEs to hemocytes and the defense functions of giant freshwater prawn (Macrobrachium rosenbergii), including hemocytic adhesion, pseudopodia formation, phenoloxidase (PO) activity, and superoxide anion (O(2)(-)) production, by means of in vitro exposure experiments. After hemocytes were treated separately with eight PAEs at concentrations of 100 microg/ml, the results showed that two PAEs (dipropyl phthalate, DPrP and diethyl phthalate, DEP) increased cells with pseudopodia formation, but decreased adhesive cells; reduction in the percentages of both pseudopodia formation and adhesive cells were detected in the dihexyl phthalate (DHP) and diphenyl phthalate (DPP) experiment groups; and di-(2-ethyl hexyl) phthalate (DEHP) decreased pseudopodia formation, but did not affect the adhesion. In addition, both PO activity and O(2)(-) production were decreased after hemocytes were treated with five PAEs (benzyl butyl phthalate (BBP), di-n butyl phthalate (DBP), DEP, DHP and DPrP), respectively. At the same time, microscopy showed that both DPrP and DHP altered morphology of the cell nucleus and led to the presence of vacuoles in cytosol of hemocytes. Using the annexin assay, and after analysis of DNA fragmentation and transmission electron microscopy (TEM), it was found that hemocytes exposed to DHP and DPrP for more than 10 min would primarily die via apoptosis, the fatality correlates with increasing treatment time; and hemocytes treated with either BBP, dicyclohexyl phthalate (DCP), DEP or DPP would primarily die via necrosis. According to these results, we suggest that all eight PAEs examined could damage hemocytes and further influence the defense mechanism of prawns. This study reveals an important precaution for prawn cultivation. PMID- 12763674 TI - Combined effects of copper and ultraviolet radiation on a microscopic green alga in natural soft lake waters of varying dissolved organic carbon content. AB - Selenastrum capricornutum was grown in two lake waters of differing dissolved organic carbon content (1.8 vs. 9.1 mg DOCl(-1)) to determine the responses of population dynamics and photosynthesis to Cu, and to assess the modifying effects of varying ultraviolet radiation (UVR) exposure. In the absence of UVR, the mean EC(50) for Cu effect on population growth rate was 2.3-2.6 microg l(-1) in the low DOC water and 17.4-26.2 microg l(-1) in the high DOC water. The variable chlorophyll a fluorescence ratio, F(v)/F(m), decreased approximately in parallel with the diminished growth rates. Exposure of the higher DOC lake water to full spectrum artificial radiation caused an increase of Cu(2+) concentration, compared to samples held in darkness or in photosynthetically active radiation (PAR) only. Full spectrum exposures also resulted in a lower (although not significantly so) EC(50) for Cu effect on growth rate, consistent with response to the moderately elevated Cu(2+) concentration. Cu(2+) concentration was unaffected by radiation exposure in the low DOC water, and EC(50)s for growth were also unaffected except in the most severe UVR treatment, which was >40% inhibited even in the absence of added Cu. Using F(v)/F(m) as an end-point, there was no evidence of interactions between UVR and Cu under the relatively low PAR exposures used here. Algal growth and photosynthesis was extremely sensitive to Cu in these soft lake waters, with EC(50)s close to current water quality standards in the low DOC water. PMID- 12763675 TI - Biotransformation and bioconcentration of pyrene in Daphnia magna. AB - Water fleas (Daphnia magna) were exposed to [14C]pyrene in the presence and absence of piperonyl butoxide (PBO), a general cytochrome P450 (CYP) inhibitor, in organic carbon-free artificial freshwater (AFW, DOC<0.2 mg l(-1)) and in natural lake water (DOC=19.9 mg l(-1)) for 24 h. The bioconcentration of total radioactivity after 24 h exposure was 50% lower in the natural lake water, indicating decreased bioavailability of pyrene by the dissolved organic matter. However, the proportions of parent compound were only ca. 12 and 19% of the total body burden in daphnids exposed in AFW and natural lake water, respectively. Therefore, the tissue concentration of the parent pyrene was not significantly different in the daphnids exposed in the two different waters. Due to extensive biotransformation the bioconcentration factor (BCF) of parent pyrene was only 16 and 23% of the BCF calculated on the basis of total radioactivity in the daphnids in AFW and natural lake water, respectively. The proportion of parent pyrene was significantly higher (over 60%) in the daphnids exposed simultaneously to PBO, which indicates the involvement of CYP monooxygenases in the biotransformation. Furthermore, increasing PBO concentration decreased the accumulation of total radioactivity in AFW but not in the natural lake water. The data demonstrate capability and importance of CYP monooxygenases in biotransformation of polycyclic aromatic hydrocarbons in D. magna. PMID- 12763676 TI - Effect of orthosilicic acid on the accumulation of trace metals by the pond snail Lymnaea stagnalis. AB - Silicon (Si) has a marked affinity for aluminium (Al(III)), but not other trace metals such as cadmium (Cd(II)) and zinc (Zn(II)). Exogenous orthosilicic acid (Si(OH)(4)) ameliorates the toxicity of Al(III) to the pond snail Lymnaea stagnalis, but its mechanism of action is unclear. Here, studies were conducted to ascertain whether interaction between orthosilicic acid and Al(III) occurs in the water column to prevent Al(III) uptake, or in the tissues to reduce the toxicity of accumulated metal. Silicon did not reduce the accumulation of Al(III) by the digestive gland (the main "sink" for trace metals in L. stagnalis) following exposure of the snail for 30 days to 500 microg l(-1) added Al(III) and 13-fold molar excess of orthosilicic acid. However, Si concentrations correlated well with Al(III) levels in the digestive gland (R(2)=0.77), giving a ratio of 2.5:1 (Al(III):Si). Exposure to Zn(II) or Cd(II) and 13-fold molar excess of orthosilicic acid did not prevent uptake of these metals, or result in a correlation between metal and Si concentrations of the snail digestive gland. These data show that aquated orthosilicic acid does not prevent Al(III) accumulation by L. stagnalis. However, following exposure, the ratio of Al(III) to Si in the digestive gland is suggestive of the early formation of hydroxyaluminosilicates, probably proto-imogolites (2-3:1 Al(III):Si). Whether hydroxyaluminates are formed ex vivo in the water column and taken up by snails into the digestive gland, or formed in situ within the digestive gland remains to be established. Either way, orthosilicic acid clearly prevents the in vivo toxicity of Al(III) rather than reducing its uptake. Silicon appears to have an important role in the handling Al(III) by the pond snail which may also have wider relevance in understanding the role of Si in ameliorating Al(III) toxicity. PMID- 12763677 TI - INNAMORA, a European Workshop focussed on the mechanisms of innate immunity in pathogen-host interaction and their exploitation in novel mucosal immunisation strategies. PMID- 12763678 TI - Innate immunity and pathogen-host interaction. AB - The skin and contiguous mucosal surfaces define the primary locus of interaction between host and micro-organisms. In this review, we focus on the innate immune system in the mucosa, which manages to deal with invading pathogens, the mechanisms that organisms have evolved in order to circumvent this primary defensive barrier and, finally, potential therapeutic manipulation of the innate immune system that was the focus of meeting at a Euroconference/Workshop on "Novel Strategies of Mucosal Immunisation through Exploitation of Mechanisms of Innate Immunity in Pathogen-Host Interaction", which was held in Siena, Italy, November 2002. PMID- 12763679 TI - Anti-cytokine therapeutics and infections. AB - In view of the increasing use of anti-cytokine-based therapies to treat autoimmune diseases, the role of specific cytokines in host defense against infection has become a highly relevant area of investigation. There are over 300,000 patients worldwide being treated with agents that specifically block the biological activities of interleukin-1 (IL-1) or tumor necrosis factor (TNF) for reducing the severity of autoimmune diseases such as rheumatoid arthritis, Crohn's disease or psoriasis. Those patients receiving anti-TNF-alpha or IL-1 blocking therapies are treated on a chronic basis. Studies suggest that other chronic inflammatory diseases will benefit from anti-cytokine therapies. However, there is a growing body of clinical evidence that neutralization of TNF-alpha is associated with an increased risk of opportunistic infections, including mycobacterial diseases. Blockade of IL-1 activity with the IL-1 receptor antagonist (IL-1Ra) appears, at present, to be relatively safe. However, because of physician under reporting (some estimates of reporting being less than 5% of these infections), the true incidence of infections, both serious and non serious, will remain unknown. Does the increase in infections associated with anti-cytokine-based therapies come as a surprise? Of the two components of host defense, the innate and the acquired responses, which are affected by anti cytokine therapies? From a wealth of rodent studies using live infection models, the following conclusions can be drawn: (1) neutralization or gene deletion for TNF-alpha is frequently associated with reduction of host defense in models of live Gram-positive or Gram-negative infections as well as infection by intracellular microbes such as Salmonella and Listeria; (2) absence of the IL-1 receptor can also result in decreased resistance to Listeria or Gram-positive bacteria and (3) TNF-alpha and IFN-gamma are required for defense against infection caused by Mycobacterium tuberculosis. PMID- 12763680 TI - Antigen dependent and independent mechanisms that sustain serum antibody levels. AB - In this article, we review three mechanisms that contribute to sustaining serum antibody levels: (i) long-lived plasma cells; (ii) persisting or cross-reactive antigen, and (iii) polyclonal activation of all memory B cells. We also present a quantitative analysis of the secondary immune response that reveals the relative contribution of antigen induced short- and long-lived plasma cells versus polyclonal activation of all memory B cells. Antigenic boost sustains high levels of serum antibodies only for a few months, while polyclonal activation sustains low levels of protective antibodies for a human lifetime. PMID- 12763681 TI - Human natural killer cell function and their interactions with dendritic cells. AB - Natural killer (NK) cells have long been considered as "primitive" and "non specific" effector cells. However, the past 10 years have witnessed dramatic progress in our understanding of how NK cells function and their role in innate defenses. Thanks to specialized inhibitory receptors specific for MHC-class I molecules, they can sense the decrease or loss of these molecules, a typical condition of potentially dangerous cells such as tumor or virally-infected cells. NK cell triggering and lysis of these cells is mediated by several activating receptors and co-receptors that have recently been identified and cloned. While normal cells are usually resistant to the NK-mediated attack, a remarkable exception is represented by dendritic cells (DC). In their immature form (iDC), they are susceptible to NK-mediated lysis because of the expression of low levels of surface MHC-class I molecules. Since the process of DC maturation (mDC) is characterized by the surface expression of high levels of MHC-class I molecules, mDC become resistant to NK cells. Exposure to live bacteria induces rapid DC maturation and, thus, resistance to NK cells. The cross-talk between DC and NK cells is more complex and involves also a DC-dependent NK cell activation and proliferation. Thus, two important players of the innate immunity may be involved in a coordinated regulation of critical events occurring at the interface between innate and adaptive immunity. PMID- 12763682 TI - Pentraxin 3, a non-redundant soluble pattern recognition receptor involved in innate immunity. AB - Pentraxin 3 (PTX3) is the first long pentraxin identified. Long pentraxins consist of a C-terminal pentraxin domain, which has sequence similarity to C reactive protein (CRP) and serum amyloid P (SAP) component (the classic short pentraxins), and of an unrelated N-terminal portion. PTX3 is made by diverse cell types, most prominently endothelial cells, macrophages and dendritic cells, in response to primary inflammatory signals (e.g. interleukin-1 (IL-1), tumour necrosis factor (TNF), lipopolysaccharide (LPS)). It binds diverse ligands, including microbial moieties, C1q and apoptotic cells. Evidence suggests that PTX3 plays a role in the regulation of innate resistance to pathogens, inflammatory reactions, possibly clearance of self-components and female fertility. PMID- 12763683 TI - Innate immune responses support adaptive immunity: NKT cells induce B cell activation. AB - Invariant NKT cells are a peculiar subset of T lymphocytes whose features, highly conserved both in the mouse and the human system, strongly recall those of other "innate lymphocytes". Following recognition of CD1d-presented glycosphingolipid antigens invariant NKT promptly release high amount of diverse cytokines concurring to the activation of the actors of both innate and acquired immune responses. For this reason, in recent years NKT cells have been the object of intensive study, aimed to understand their role in diverse patho-physiological conditions and to exploit the possibility to take advantage of their "adjuvant like" activity in the formulation of new vaccines. As antibodies are an essential part of many immune responses, we focused our attention on invariant NKT-B cell interactions analyzing their influences on B cell activation and effector functions. The results of this study demonstrate that human invariant NKT cells can provide direct help for B cell proliferation and antibody production through CD1d-restricted mechanisms. Remarkably, help to B lymphocytes by invariant NKT cells is delivered also in the absence of exogenous antigen, suggesting the existence of an endogenous ligand presented by CD1d on B cells. PMID- 12763684 TI - The role of Toll-like receptor 2 in microbial disease and immunity. AB - Cells expressing Toll-like receptor (TLR), TLR2 in association with TLR1, TLR6 or some other unknown co-receptor can respond upon interaction with a large variety of microbial ligands. The variety of TLR2 ligands is the greatest among all the TLRs and this is due to the heterodimerization needed for TLR2 mediated responses. Like other TLRs, TLR2 signaling induces antigen presenting cell activation, pro-inflammatory cytokine production and increased expression of co stimulatory ligand expression. These events are important for induction of innate immune responses and improved acquired immunity. There is strong suggestive evidence that alteration or lack of TLR2 function in vivo may correlate to decreased immune protection from pathogens that contain TLR2 ligands, but more work needs to be performed to strengthen this correlation. PMID- 12763685 TI - TLR-induced negative regulatory circuits: role of suppressor of cytokine signaling (SOCS) proteins in innate immunity. AB - Recognition of pathogen-associated molecular pattern (PAMP) by PAMP receptors of innate immune cells leads to activation and initiation of innate and adaptive immune responses. While the initial steps have been elucidated recently, the mechanisms how innate immune responses are controlled and regulated are only poorly understood. Suppressor of cytokine signaling (SOCS) proteins seem to play an important role in this process. It has been recognized that SOCS proteins not only regulate JAK/STAT-dependent cytokine signaling pathways but are also induced by TLR stimuli and in addition might also regulate TLR signaling. PMID- 12763686 TI - Innate and adaptive mucosal immunity in protection against HIV infection. AB - Control of the HIV pandemic requires an effective vaccine. The difficulties in developing a preventive vaccine are generally believed to be due to the rapid rate of mutation of HIV that escapes cytotoxic lymphocytes (CTL) and the problems in induction of neutralising antibodies to wild strains of HIV. These difficulties should re-orientate vaccine strategy into four somewhat neglected areas of immunisation. Innate immunity, with its rapid protective response to infection that is independent of memory and relies on an optimal mucosal adjuvant. Targeting the genital and rectal mucosa, with the associated lymph nodes, as an immune response has to be elicited directly on encountering HIV during sexual intercourse. Stimulating a broadly based adaptive immune response that enhances the memory CD4(+) and CD8(+) T cells and B cells, induces maturation of dendritic cells and results in Th1 polarised immunity. Taking advantage of "experiments of nature", by utilising host antigens, as manifested by protection against HIV infection in homozygous Delta32 CCR5 individuals and in allo-immunity. PMID- 12763688 TI - Vaccination strategies. An overview. AB - Existing vaccines have contributed significantly to the reduction of the mortality and morbidity burdens of many infectious diseases. In many instances, however, the development of these vaccines has been empirical. Furthermore, the impressive progress in the field of vaccines has been mainly driven the progress in molecular biology and microbiology more than by the progress in immunobiology. Nevertheless, the new challenges vaccinology is facing nowadays can be approached through a comprehensive understanding of the mechanisms behind the induction and the maintenance of efficacious immune responses triggered by vaccines. The new vaccination strategies aim at exploiting the new knowledge in the field of dendritic cells (DC; and their role in priming immune responses), in the field of antigen processing and presentation for the generation of antigen-specific cytolytic T cells, and in the field of induction and maintenance of immunological memory, with the ultimate goal of developing better vaccines with an enhanced safety and efficacy not only in children and adults, but also at neonatal age. PMID- 12763687 TI - Role of complement in the control of HIV dynamics and pathogenesis. AB - In all ex vivo preparations of HIV tested so far, C3 fragments and, after seroconversion, antibodies were detected on the viral surface. This indicates that HIV survives complement-mediated lysis. The virus has adopted different protection mechanisms to keep complement activation under the threshold necessary to induce virolysis. Among them are complement regulatory proteins that remain functionally active on the surface of HIV and turn down the complement cascade and serum proteins with complement regulatory activities. Therefore, opsonized virions accumulate in HIV-infected individuals, and subsequently adhere to complement receptor (CR) expressing cells. Among them are B cells, which bind opsonized virus. Such bound virus is efficiently transferred to autologous T cells, which subsequently are infected. Other cells interacting via CR with opsonized HIV are follicular dendritic cells (FDC). As shown by ex vivo experiments, up to 80% of virus is bound to follicular dendritic cells through C3 CR interactions. In the brain, HIV is not only interacting with complement proteins, but is able to induce their expression. Thus, interaction of HIV with the complement system is a main mechanism for pathogenesis to AIDS, since retention of (complement-resistant) opsonized viral particles on cell surfaces via CRs occurs in different compartments in HIV-infected individuals, thereby promoting transmission of virus to other permissive cells. PMID- 12763690 TI - Bacillus spores for vaccine delivery. AB - Spores of the genus Bacillus have been used for a long time as probiotics for oral bacteriotherapy both in humans and in animals. Spores are also employed in a veterinary vaccine against anthrax. Despite this long lasting and extensive use, the specific contribution of spores to the beneficial effects of probiotics and to the immunogenicity of the vaccine is not completely elucidated. This review focuses on the different aspects of the use of spore preparations. In particular the use of recombinant spores as vaccine delivery vehicles is described and discussed. PMID- 12763689 TI - Mucosal immunisation and adjuvants: a brief overview of recent advances and challenges. AB - Mucosal immunisation may be used both to prevent mucosal infections through the activation of anti-microbial immunity and to treat selected autoimmune, allergic or infectious-immunopathological disorders through the induction of antigen specific tolerance. The development of mucosal vaccines, whether for prevention of infectious diseases or for immunotherapy, requires antigen delivery and adjuvant systems that can efficiently help to present vaccine or immunotherapy antigens to the mucosal immune system. Promising advances have recently been made in the design of more efficient mucosal adjuvants based on detoxified bacterial toxin derivatives or CpG motif-containing DNA, and perhaps even more striking progress has been done in the use of virus-like particles as mucosal delivery systems for vaccines and of cholera toxin B subunit as antigen vector for immunotherapeutic tolerance induction. However, it is a memento that two recently developed mucosal vaccines for human use against rotavirus diarrhoea and influenza were withdrawn after a short period in the market because of adverse reactions among the vaccinees, thus emphasising the difficult and challenging task also for mucosal immunisation of combining vaccine and adjuvant efficacy with safety and acceptability. PMID- 12763691 TI - Induction of immune responses by attenuated isogenic mutant strains of Listeria monocytogenes. AB - We have generated isogenic Listeria monocytogenes mutant strains to study the induction of protective immunity in mice. These strains harbored either a specific deletion within the actin nucleator (actA) and/or have multiple deletions within the actA and phospholipase B (plcB) genes. In comparison to the wild type parental L. monocytogenes EGDe strains, the mutant strains were extremely low in virulence and were rapidly eliminated by the host during the first days of infection. Nevertheless, a single immunization with both mutant strains (EGDe DeltaactA2 and DeltaactADeltaplcB) efficiently induced and maintained effector memory (CD8(+)) T cells and has provided animals with a state of long-lasting protective immunity against wild type L. monocytogenes. These mutant strains can be used as live vaccines against the corresponding virulent pathogen and as carriers for introducing heterologous protective antigens into animals and humans. PMID- 12763692 TI - The value of vaccines. AB - Today, we have the technology to make vaccines against most infectious diseases and in theory we could free mankind from most of them. In spite of the great progress of science and technology, vaccines are an endangered species and there are increasing non-technological barriers to their development. Indeed, we have no mechanisms for developing vaccines needed only in developing countries, and in developed countries they are not a priority. Industry is walking away from vaccines and even the existing ones are in jeopardy. The reasons for the low interest in vaccines lie in the high risk and low profitability of the vaccine business. A story about the consequences that an infectious disease had on the economic development of the city of Siena in 1348 is used to show that our society is not calculating the intangible values deriving from vaccination. The failure of assigning the right value to vaccines and preventive medicine is a major risk of today's world that, having the opportunity of improving the proportion of healthy population, may have made the choice of increasing the number of chronically sick people. PMID- 12763693 TI - Thinking downstream to accelerate the introduction of new vaccines for developing countries. AB - Introduction of new vaccines into public health programs in developing countries requires 'translational research', initiated even during the phase of vaccine development, to generate the evidence base necessary for rational public health decision-making. Translational research encompasses assessments of the local disease burden, demonstration projects of vaccines in realistic public health programs, analysis of the economic impact of vaccine introduction, assessment of community and policy-maker perceptions about the importance of a disease and the need to vaccinate against it, and identification of distribution channels and financial mechanisms for deploying and paying for a new vaccine. In the absence of this background information, even a successful vaccine may risk substantial difficulties and delays in its introduction into programs for the poor in developing countries. PMID- 12763694 TI - The European research effort for HIV/AIDS, malaria and tuberculosis. AB - The global emergency caused by HIV/AIDS, malaria and tuberculosis requires for new approaches and actions to confront these three major poverty-related diseases. In response to this emergency, the European Commission provides a broad comprehensive approach in a wide range of policy areas, including trade, development and research. For research, the overall strategy is to develop new drugs, vaccines or other effective interventions by two main mechanisms: (i) support of research projects for the development of new promising candidates through pre-clinical and early human testing and (ii) establishment of a programme to support phases II and III clinical trials in Africa. The Sixth Framework Programme (FP6) (2002-2006) allocates a total of 400 million euro to research on HIV/AIDS, malaria and tuberculosis with about 200 million for each of the two interlinked components. Research projects, aiming at developing new promising candidates, should create large consortia capable of integrating different approaches and disciplines providing the necessary critical mass to test and compare different scientific ideas. Projects should cover different phases in the development process ranging from basic knowledge generated from genomics or immunology to pre-clinical testing in animal models and finally validation in safety trials. The new instruments, mainly Integrated Projects and Network of Excellence, are the preferred means to implement the proposed approach. The European and Developing Countries clinical trials partnership (EDCTP) will help to overcome the bottleneck of demonstrating a proof of principle for promising vaccine or drug candidates in testing them in early efficacy trials in endemic areas, particularly in sub-Saharan Africa. PMID- 12763695 TI - Novel human vaccine strategies and the 5th Framework Programme: pushing the envelope. AB - Mucosal vaccines could result in a great scientific and practical achievement. More than three decades of research in experimental models have shown promising results in stimulating mucosal immune responses, thus, it was expected that within a short time mucosal vaccines for human use could be achieved. Indeed this is not being the case. In the last few years, the most important oral vaccine, the anti-polio developed by Sabin in the fifties, has been progressively abandoned in developed countries to avoid the few cases of disease caused by the vaccine. Furthermore, two recently developed mucosal vaccines for human use against rotavirus diarrhoea and influenza were withdrawn after a short period in the market because of adverse reactions among the vaccinees. This controversial situation has created a difficult future for research on mucosal vaccine at the industrial level. A great help and encouragement for believers in mucosal vaccines has been given by the EU Commission through the 5th Framework Programme (5FP). At the end of the first projects of the 5FP, it is quite clear that mucosal vaccines are experiencing a real renaissance. The Euroconference/Workshop "Novel Strategies of Mucosal Immunisation through Exploitation of Mechanisms of Innate Immunity in Pathogen-Host Interaction", organised under the sponsorship of the EU Commission and reported in this special issue of Vaccine, witnesses a very creative moment of European groups involved in mucosal immunology. This conclusive paper of the issue is intended to describe a positive experience of some European scientists that have been working together in organised fashion within two EU projects. The first, defined by the acronym MUCIMM, was aimed to pave the way to tackle mucosal vaccines with different approaches, mainly that of new delivery systems and adjuvants, that of dissecting the fine mechanisms of basic mucosal responses and that of obtaining meaningful assays to measure human immune responses to mucosal vaccines. The second, the MUCADJ project, was aimed to prove that an intranasally delivered influenza vaccine induces protective levels of immunity in human adult volunteers. The results obtained demonstrate that mucosal vaccines for humans are feasible. It is interesting to note how this model of making biomedical research is flourishing, becoming an example for work organisation. At the same time it is important to underline some of the limits of this cooperative approach and it is also mandatory to spend a word of consciousness regarding the impact that the new European regulations could exert on mucosal vaccines, particularly for those based on live delivery systems. In a world where the damages caused by syringe needles are still largely visible, mucosal vaccines could represent an extremely important tool to fight infections, particularly in the large population of the most impoverished. The EU Commission will still encourage and support mucosal vaccines in the 6FP. The scientific community must decide in which direction has to proceed to avoid regulatory problems that could halt a promising tool to improve human health. The envelope has been pushed, the trip begins. PMID- 12763696 TI - Being inconsistent about consistency: when coefficient alpha does and doesn't matter. AB - One of the central tenets of classical test theory is that scales should have a high degree of internal consistency, as evidenced by Cronbach's a, the mean interitem correlation, and a strong first component. However, there are many instances in which this rule does not apply. Following Bollen and Lennox (1991), I differentiate between questionnaires such as anxiety or depression inventories, which are composed of items that are manifestations of an underlying hypothetical construct (i.e., where the items are called effect indicators) and those such as Scale 6 of the Minnesota Multiphasic Personality Inventory (Hathaway & McKinley, 1943) and ones used to tap quality of life or activities of daily living in which the items or subscales themselves define the construct (these items are called causal indicators). Questionnaires of the first sort, which are referred to as scales in this article, meet the criteria of classical test theory, whereas the second type, which are called indexes here, do not. I discuss the implications of this difference for how items are selected, the relationship among the items, and the statistics that should and should not be used in establishing the reliability of the scale or index. PMID- 12763697 TI - Discontinuities, side steps, and finding a proper place: an autobiographical account. AB - In this invited autobiography, I present an account of some influential events and experiences in my life that led me eventually, after some false starts, to a career in personality assessment psychology. The psychological research and applied areas (self-report personality assessment and cross-cultural research) with which I have been identified for over 40 years can, however, be seen in early interests, although their impact and their direction were not always apparent at the time of occurrence. Several people who were influential in my career development are noted and a number of important collaborators who helped sustain my efforts in personality assessment research and practice over the years are highlighted. PMID- 12763698 TI - Parental ratings of son's behavior problems in high-risk families: convergent validity, internal structure, and interparent agreement. AB - In this research, we examined the convergent validity, internal structure, and interparent agreement of the Child Behavior Checklist (CBCL; Achenbach, 1991a) using data from a community sample of 317 high-risk families. The sample consisted of 218 families with alcoholism (biological mother, father, and son) and 99 matched comparison families without alcoholism; all families were intact and included a 3- to 5-year-old son at first contact. Evidence was found for the convergent validity of the CBCL. In addition, although interparent agreement was low, the internal structure was confirmed, and structural invariance was high between mothers and fathers and in families with and without alcoholism. The effects of parental psychological and cognitive functioning on parent agreement were examined and found to have a minimal effect on parent agreement. These results demonstrate the validity of the CBCL for use by parent raters with alcoholism and their sons and emphasize the importance of including both versus only 1 parent in research using the CBCL. PMID- 12763699 TI - Factorial validity of the Conners' Parent Rating Scale-revised: short form with psychiatric outpatients. AB - To assess the factorial validity of the 27-item Conners' Parent Rating Scale Revised: Short Form (CPRS-R:S; Conners, 1997), 100 (50%) male and 100 (50%) female psychiatric outpatients between 5 and 16 years old were rated by a parent. A confirmatory factor analysis of the 18 item ratings from the CPRS-R:S Oppositional, Cognitive Problems, and Hyperactivity scales provided only tentative support for scoring these scales as Conners (1997) recommended. However, an exploratory principal-axis factor analysis with all 27 item ratings found 2 dimensions: 1 dimension was composed of the 6 items in the Oppositional scale, and other dimension contained the remaining 21 items. An attention deficit hyperactivity disorder (ADHD) Total Symptoms scale was constructed by summing the ratings for these 21 symptom ratings, and this scale was found to be as effective as the Hyperactivity scale was in discriminating between youth who were and were not eventually diagnosed with an ADHD. We discuss the results as providing an alternate way of scoring the CPRS-R:S to screen for an ADHD in child and adolescent psychiatric outpatients. PMID- 12763700 TI - A critique of Lilienfeld et al.'s (2000) "The scientific status of projective techniques". AB - Lilienfeld, Wood, and Garb (2000) published a largely negative critique of the validity and reliability of projective methods, concentrating on the Comprehensive System for the Rorschach (Exner, 1993), 3 systems for coding the Thematic Apperception Test (TAT; Murray, 1943) cards, and human figure drawings. This article is an effort to document and correct what I perceive as errors of omission and commission in the Lilienfeld et al. article. When projective measures are viewed in the light of these corrections, the evidence for the validity and clinical usefulness of the Rorschach and TAT methods is more robust than Lilienfeld et al. represented. PMID- 12763701 TI - Temporal stability in the Rorschach method: a meta-analytic review. AB - The temporal stabilities of the Rorschach method scoring systems were investigated. A comprehensive literature search was conducted to find all test retest studies, both regular and from control samples in therapy evaluation studies. Meta-analyses were conducted for samples, individual entries, and variables in 36 samples. Separate analyses were made for 10 samples using the Comprehensive System (CS; Exner, 1993). The temporal stability was high, especially those obtained from regression model predictions. The CS consistently shows higher stability than other systems, whereas increasing retest intervals leads to decreasing stability. Shortcomings of the available studies are highlighted and discussed, and recommendations are given for design and methodology. PMID- 12763702 TI - Longitudinal hierarchical linear modeling analyses of California Psychological Inventory data from age 33 to 75: an examination of stability and change in adult personality. AB - Twenty aspects of personality assessed via the California Psychological Inventory (CPI; Gough & Bradley, 1996) from age 33 to 75 were examined in a sample of 279 individuals. Oakland Growth Study and Berkeley Guidance Study members completed the CPI a maximum of 4 times. We used longitudinal hierarchical linear modeling (HLM) to ask the following: Which personality characteristics change and which do not? Five CPI scales showed uniform lack of change, 2 showed heterogeneous change giving an averaged lack of change, 4 showed linear increases with age, 2 showed linear decreases with age, 4 showed gender or sample differences in linear change, 1 showed a quadratic peak, and 2 showed a quadratic nadir. The utility of HLM becomes apparent in portraying the complexity of personality change and stability. PMID- 12763703 TI - Incremental validity of the MMPI-A content scales in the prediction of self reported symptoms. AB - The incremental contribution of the MMPI-A (Butcher et al., 1992) content scales to the prediction of scores on self-report measures of psychopathology was examined in a sample of 62 adolescents in inpatient treatment and 59 adolescents from the community. All participants completed the MMPI-A and a battery of criterion measures. A series of hierarchical regression analyses was conducted in which the MMPI-A clinical and content scales served as the independent variables and the criterion measures as the dependent variables. The content scales were found to have incremental validity beyond the clinical scales in predicting variance in the criterion measures. Similarly, the clinical scales also demonstrated incremental validity over the content scales in making these predictions. Both sets of scales made independent contributions to the prediction of sample membership (clinical vs. nonclinical). Findings suggest that both the clinical and content scales of the MMPI-A make significant contributions to the assessment of adolescents' psychological functioning. PMID- 12763704 TI - Gender differences in the psychosocial and cognitive correlates of physical activity among Taiwanese adolescents: a structural equation modeling approach. AB - This study examined gender differences in the factors related to physical activity in 832 Taiwanese adolescents. Differences in psychosocial and cognitive correlates were noted by gender group. Taiwanese adolescent girls reported lower physical activity self-efficacy and less perceived benefits and more perceived barriers to being active than boys. Girls compared to boys reported significantly more positive social support, modeling, and norms from parents to be active but significantly less social support and norms from their peers. Structural equation modeling was used to test the direct and indirect paths of a model of proposed correlates and physical activity. The results indicated that peer influences had both direct and indirect paths to physical activity for both genders. Among all examined variables, perceived self-efficacy was the strongest correlate of physical activity for these adolescents. The findings of this study provide information relevant to designing physical activity interventions targeted to Taiwanese adolescents. PMID- 12763705 TI - Affects and autonomic cardiac reactivity during experimentally induced stress as related to precursors of insulin resistance syndrome. AB - This study examined the association of insulin resistance syndrome (IRS) precursors with state affects and autonomic reactivity in randomly selected healthy young adults. It was asked whether IRS precursors are able to predict a person's mental and physiological coping with acute stress over an 11-year follow up period. IRS parameters were serum insulin, high-density lipoprotein cholesterol, triglycerides, systolic blood pressure, body-mass index, and subscapular skinfold thickness. In the psychophysiological experiment, state affects and cardiac responses (heart rate, respiratory sinus arrhythmia, and pre ejection period) were measured during different challenges. The main result was that IRS precursors predicted high levels of negative emotions during the challenges; IRS accounted for 19% of the variance in tiredness. IRS was unrelated to cardiac reactivity. Mechanisms underlying the associations found are discussed. PMID- 12763706 TI - Association between job characteristics and health behaviors in Japanese rural workers. AB - Associations between job characteristics defined by the Karasek's job demand control model and health behaviors were investigated in a cross-sectional analysis of 6,759 Japanese rural workers. High psychological demands were associated with heavy smoking, exaggerated prevalence of alcohol drinking, and high work-related physical activity. Low job control was associated with lower consumption of vegetables, a smaller quantity number of cigarettes smoked, and a low level of work-related physical activity. Job strain, a combined measure obtained from the ratio of demands to control, was associated with lower vegetable consumption, low prevalence of smoking, and high prevalence of current alcohol drinking. Stratified analyses by occupations and gender provided some but not all of the explanations for the unexpected findings. The results indicate a possible association between psychosocial job characteristics and health behaviors. A few unexpected findings prevent complete support to one of the hypothetical pathways that the job characteristics lead to cardiovascular disease through behaviors. PMID- 12763707 TI - Managing urinary incontinence across the lifespan. AB - In the 1996 baseline surveys of the Australian Longitudinal Study of Women's Health (ALSWH), 36.1% of mid-age women (45-50) and 35% of older women (70-75) reported leaking urine. This study aimed to investigate (a) the range of self management strategies used to deal with urinary incontinence (UI); (b) the reasons why many women who report leaking urine do not seek help for UI; and (c) the types of health professionals consulted and treatment provided, and perceptions of satisfaction with these, among a sample of women in each age group who reported leaking urine "often" at baseline. Five hundred participants were randomly selected from women in each of the mid-age and older cohorts of the ALSWH who had reported leaking urine "often" in a previous survey. Details about UI (frequency, severity, and situations), self-management behaviors and help seeking for UI, types of health professional consulted, recommended treatment for the problem, and satisfaction with the service provided by health care professionals and the outcomes of recommended treatments were sought through a self-report mailed follow-up survey. Most respondents had leaked urine in the last month (94% and 91% of mid-age and older women, respectively), and 72.2% and 73.1% of mid-aged and older women, respectively, had sought help or advice about their UI. In both age groups, the likelihood of having sought help significantly increased with severity of incontinence. The most common reasons for not seeking help were that the women felt they could manage the problem themselves or they did not consider it to be a problem. Many women in both cohorts had employed avoidance techniques in an attempt to prevent leaking urine, including reducing their liquid consumption, going to the toilet "just in case," and rushing to the toilet the minute they felt the need to. PMID- 12763709 TI - Assessment of muscular tension in children. AB - The objective of this study was to test the reliability and validity of a semistructured questionnaire concerning children's experience of muscular tension and its relation to daily life experiences and pain. The reliability of the questions were tested by the test-retest method, and the answers were validated in a sample of children. The answers in the test-retest procedure were the same or had virtually the same content in 8 out of 9 children (k = 0.80). When asked about the sensation of bodily tension, 38 out of 42 children, aged 7 to 15, said that they had experienced this sensation, and 32 of them could give a more detailed description of it using words like jittery, stiff, tight, and unpleasant. These 32 children could also describe where in the body they felt the tension: Abdomen, head, legs, and arms were common locations. Thirty-six children described how the tension was triggered by life experiences that had to do with stress. In 14 of the 42 children, the tension induced pain in the areas of the body where they experienced tension, such as the stomach and the head. Thus, in a small sample of children, a semistructured questionnaire showed acceptable reliability and validity. This indicates, preliminarily, that children over 6 years of age can identify the sensation, location, and inducing factors of bodily tension and its relation to pain and describe these experiences in personal words. PMID- 12763708 TI - Examining the influence of biological and psychological factors on cognitive performance in chronic fatigue syndrome: a randomized, double-blind, placebo controlled, crossover study. AB - The pathophysiology of chronic fatigue syndrome (CFS) remains unclear; however, both biological and psychological factors have been implicated in establishing or maintaining this condition. People with CFS report significant and disabling cognitive difficulties such as impaired concentration that in some cases are exacerbated by exposure to chemical triggers. The aim of this study was to determine if neuropsychological deficits in CFS are triggered by exposure to chemicals, or perceptions about the properties of these substances. Participants were 36 people with a primary diagnosis of CFS, defined according to Centers for Disease Control (CDC) criteria. A randomized, double-blind, placebo-controlled, crossover design was used, with objective assessment of neuropsychological function and participant rating of substance type, before and after exposure to placebo or chemical trigger. Results showed decrements in neuropsychological tests scores on three out of four outcome measures when participants rated the substance they had been exposed to as "chemical." No change in performance was found based on actual substance type. These results suggest that cognitive attributions about exposure substances in people with CFS may be associated with worse performance on neuropsychological tasks. In addition, these findings suggest that psychological interventions aimed at modifying substance-related cognitions may reduce some symptoms of CFS. PMID- 12763710 TI - Age and musculoskeletal pain. AB - Using questionnaires, we analyzed associations between different pain variables (e.g., pain intensity) and age (20-65+ years) among 949 primary pain patients. Older patients (a) were more often divorced, were blue-collar workers, were less educated, and had greater difficulties with living expenses; (b) had pain of longer duration, more frequently and of more complexity, and felt more disabled; (c) consumed more painkillers, analgesics, sedatives, and other medications, and had received more pain treatments; and (d) had more health problems. Younger patients had more severe pain, were financially strained, and were more often unemployed. A multivariate regression analysis showed that high disability was more determined by older than young age. However, other factors (e.g., pain complexity) were also important. Thus, older and younger patients experienced their pain differently PMID- 12763711 TI - Interactive relation of insulin and gender to cardiovascular reactivity in healthy young adults. AB - High levels of insulin may promote hypertension pathogenesis, in part, via enhanced sympathetic nervous system (SNS) activity. This study examined potential interactive relations of fasting insulin levels, gender, and race to cardiovascular reactivity-a correlate of SNS activation. Hemodynamic responses to 4 laboratory challenges were determined by impedance cardiography in 64 healthy young adults (ages 18-26; 48% male; 50% White, 50% African American). Also examined were lipoprotein lipids, central and total adiposity, self-reported dietary factors, and physical activity. High-insulin (>10.2 mU/ml) men showed greater total peripheral resistance and longer pre-ejection period responses than low-insulin ( pound 10.2 mU/ml) men. High-insulin women displayed greater cardiac index responses than high-insulin men. High insulin levels were related to greater percentage body fat, dietary carbohydrate and fat intake, lower high density lipoprotein (HDL) cholesterol (in men), higher total cholesterol (in women), and a trend toward higher triglycerides. Cardiovascular reactivity findings were unchanged after statistical adjustment for total and HDL cholesterol, triglycerides, percentage body fat, dietary carbohydrates, and fat. The Gender x Insulin (continuous scores) interaction accounted for 7% and 9% of the variance in cardiac index and total peripheral resistance responses, respectively. These results indicate that high insulin levels are associated with greater vascular reactivity in young men and cardiac reactivity in young women. Enhanced cardiovascular reactivity may constitute a biobehavioral dimension of the metabolic syndrome. PMID- 12763712 TI - Depression and poverty among African American women at risk for type 2 diabetes. AB - Poverty is associated with negative health outcomes, including depression. Little is known about the specific elements of poverty that contribute to depression, particularly among African American women at risk for type 2 diabetes. This study examined the relationships of economic and social resources to depression among African American women at high risk for the development of type 2 diabetes (N = 181) using the Conservation of Resources theory as a conceptual framework. Women were assessed at 3 time points in conjunction with a dietary change intervention. At baseline, 40% of women reported clinically significant depression, and 43.3% were below the poverty line. Depressed women reported fewer economic assets and greater economic distress than nondepressed peers. Multivariate logistic regression analyses indicated that nonwork status, lack of home ownership, low appraisal of one's economic situation, low self-esteem, and increased life events were significantly associated with depression at baseline. Longitudinal multivariate logistic regression models indicated that income, home ownership, future economic appraisal, life events, and self-esteem predicted depression trajectories at Time 3. These results speak to the multifaceted sources of stress in the lives of poor African American women. Interventions that address the economic and social factors associated with depression are needed. PMID- 12763713 TI - Problem solving in diabetes self-management: a model of chronic illness self management behavior. AB - Management of chronic illnesses is a significant public health concern that is made more challenging by problems of regimen adherence. Effective self-management of chronic illnesses such as diabetes requires not only technical skill to perform regimen behaviors but also problem-solving skills to manage daily barriers to regimen adherence and to make appropriate adjustments to the self care regimen. A review of the empirical literature on the relation between problem solving and disease self-management in diabetes, a chronic illness exemplar, illuminates methodological limitations that indicate a need for a theoretical framework for problem solving applied to chronic disease self management. A problem-solving model of chronic disease self-management is proposed, derived from theories of problem solving in cognitive psychology, educational/learning theory, and social problem solving. Four essential components of problem solving in disease self-management are identified: (a). problem-solving skill, (b). problem-solving orientation, (c). disease-specific knowledge, and (d). transfer of past experience. The model is illustrated, and empirical support for this problem-solving model in the diabetes literature is discussed. The model has utility in driving testable hypotheses regarding the relation of disease-specific problem solving to chronic illness management, in developing problem-solving assessment instruments relevant to disease self management, and in guiding disease self-management training and interventions. PMID- 12763714 TI - Population-based randomized controlled trial of a stage-targeted physical activity intervention. AB - BACKGROUND: Intervention trials with self-selected participants have shown that mailed stage-targeted print materials can increase participation in physical activity in the short term. We examined the effects of a mailed stage-targeted print intervention designed to promote physical activity, in a random sample of adults living in a regional city. METHOD: Participants (n = 462, 40-60 years of age) were randomly allocated to an intervention (n = 227) or control group (n = 235). Measures included validated 2-week physical activity recall and stage of motivational readiness for physical activity. The intervention consisted of a single mailing of a letter and full-color stage-targeted booklets (specific to precontemplation, contemplation, preparation, and action/maintenance) 1 week postbaseline. Follow-up interviews were conducted at 2 and 6 months postbaseline. RESULTS: After 2 months, participants in the intervention group were significantly more likely to meet the current American College of Sports Medicine/Centers for Disease Control and Prevention recommendation for sufficient physical activity than those in the control group (adjusted odds ratio [OR] = 2.40; 95% confidence interval [CI] = 1.44-3.99). After 6 months, intervention participants who reported receiving and reading the intervention materials were significantly more likely to be meeting the sufficient physical activity criterion compared with the control group (adjusted OR = 2.03; 95% CI = 1.16 3.56). CONCLUSIONS: The stage-targeted print intervention was effective in promoting short-term increases in physical activity and was most effective for participants who recognized and used the materials. This low-cost, generalizable intervention has demonstrated potential as a practical population-based physical activity promotion strategy. Further research is required before widespread dissemination would be justified, as additional strategies may be required to ensure sustained change. PMID- 12763715 TI - Coping and psychological distress among symptomatic HIV+ men who have sex with men. AB - This study evaluated relations among indicators of latent coping factors and psychological distress while incorporating measures of life stress and HIV illness related factors simultaneously among 211 symptomatic, HIV+ men who have sex with men (MSM). Participants were all assessed at a single time point. A structural equations model with latent factors for approach-oriented coping, avoidant-oriented coping, and psychological distress showed adequate fit. Furthermore, significant associations were identified among latent factors for approach-oriented coping, avoidance coping, and psychological distress; specifically, greater use of approach-oriented coping strategies and less use of avoidant-oriented coping were associated with lower levels of psychological distress. The model was revised to incorporate variables significantly associated with psychological distress (i.e., personal loss-total events, personal loss controllability, and HIV-related symptoms). Relations among the coping and psychological distress latent factors remained significant. The results suggest that HIV+ MSM who do not have the coping skills or resources necessary to use adequate coping strategies to face the chronic burdens associated with HIV illness are likely to experience higher levels of psychological distress, independent of life stress and ongoing HIV-related symptoms. PMID- 12763716 TI - Longitudinal predictors of continued tobacco use among patients diagnosed with cancer. AB - Even though continued smoking by cancer patients adversely affects survival and quality of life, about one third of patients who smoked prior to their diagnosis continue to smoke after their diagnosis. The implementation of smoking cessation treatments for cancer patients has been slowed by the lack of data on correlates of tobacco use in this population. Thus, this longitudinal study assessed demographic, medical, addiction, and psychological predictors of tobacco use among 74 head, neck, and lung cancer patients. Multivariable binary logistic regression analyses, with outcome categorized as smoker or nonsmoker, indicated that the likelihood that patients would be a smoker was associated with lower levels of perceived risk and a higher level of quitting cons. Multivariable nominal logistic regression, with outcome classified as continuous smoker, continuous quitter, relapser, or follow-up quitter, indicated that: (a). patients categorized as continuous smokers reported significantly lower quitting self efficacy than follow-up quitters and continuous quitters, (b). relapsers reported a significantly lower level of quitting self-efficacy than either follow-up quitters or continuous quitters, and (c). continuous smokers exhibited a significantly lower level of risk perceptions than continuous abstainers. These findings can be useful for the development and evaluation of treatments to promote smoking cessation among cancer patients. PMID- 12763718 TI - Keeping surprise blood clots at bay. PMID- 12763717 TI - Avoid an aspirin roadblock. PMID- 12763719 TI - A HAART ache from HIV therapy? PMID- 12763720 TI - Enter the new Iron Age. PMID- 12763721 TI - Heart Beat: Women and heart disease. PMID- 12763722 TI - Heart Beat: Experimental treatments turn toxins into medicine. PMID- 12763723 TI - Ask the Doctor: Why didn't a stress test warn me I was about to have a heart attack? PMID- 12763724 TI - Ask the Doctor: Could chest pain be a sign that I have cancer again? PMID- 12763725 TI - Advances in reducing colorectal cancer risk. Colorectal cancer is the third most common cancer in women and the second leading cause of cancer death. These statistics belie how preventable this disease really is. PMID- 12763726 TI - Replacing worn-out hips. Decades ago, worn-out hips meant life in a wheelchair. But in the 1960's, modern materials made it possible to replace these large ball and-socket joints with mechanical implants. PMID- 12763727 TI - Celiac disease: not that rare? PMID- 12763728 TI - The minerals we need. The human body and a sturdy building have many elements- that is, minerals--in common, including calcium, iron, potassium, copper, molybdenum, and zinc. Both are built to last. PMID- 12763729 TI - By the way, doctor. I keep hearing that alcohol, even in moderate amounts, raises the risk of developing breast cancer. How does it do that? PMID- 12763731 TI - Fainting. Why people pass out--and when it's a worry. PMID- 12763730 TI - By the way, doctor. I've heard about a new rheumatoid arthritis drug that I'd only have to take every two weeks. What do you know about it? PMID- 12763732 TI - Erectile dysfunction: can it be prevented? PMID- 12763733 TI - Diet and prostate cancer: new evidence favoring fish. PMID- 12763734 TI - On call. I am a 68-year-old man with a question about my waistline. I had surgery for prostate cancer seven years ago, and my PSA has remained okay. My health is otherwise perfect. I work out at my health club every day at 6 a.m., and I eat a healthful diet. I'm six-foot-one and my weight is 180. My weight has not changed in the past 10 years, and I wear the same size 36 trousers as always, but my waist seems to bulge a bit more each year. What can I do to correct my potbelly? PMID- 12763735 TI - Confronting suicide. Part I. Vulnerable people and perilous circumstances. PMID- 12763737 TI - Alien abduction and recovered memory. PMID- 12763736 TI - Can a troubled mind spell trouble for the heart? Part II. In Part I, we reviewed the evidence that links depression to heart disease and explored some possible reasons for the connection. Here we discuss implications for treatment and research. PMID- 12763738 TI - Protecting battered women. PMID- 12763739 TI - The sadness of Alzheimer's. PMID- 12763740 TI - Seeing the face of anger. PMID- 12763741 TI - Hypertrophy changes the muscarinic receptor subtype mediating bladder contraction from M3 toward M2. AB - Major pelvic ganglion electrocautery (MPGE) and spinal cord injury in the rat induce bladder hypertrophy and a change in muscarinic receptor subtypes mediating bladder contraction from predominantly M3 to a combination of M2 and M3. To determine whether this is a result of bladder hypertrophy or denervation, we studied the following groups: sham-operated controls, urinary diversion (DIV), MPGE together with urinary diversion (DIV-DEN), bilateral MPGE (DEN), bladder outlet obstruction (BOO), and MPG decentralization (MPGDEC). The degree of bladder denervation was determined by the maximal carbachol response normalized to the response to electric field stimulation. Receptor subtype density was determined by immunoprecipitation. The affinity of subtype-selective muscarinic antagonists for inhibition of carbachol-induced contractions was used to determine the subtype-mediating contraction. DEN, MPG-DEC, and BOO bladders were hypertrophic whereas DIV bladders were atrophic compared with sham operated. Bladder contraction in sham-operated, DIV, and DIV-DEN was mediated by the M3 receptor subtype, whereas the M2 subtype participated in contraction in the DEN, MPG-DEC, and BOO groups. The hypertrophied bladders had an increase in total and M2 receptor density while all experimental groups showed a reduction in M3 receptor density. Thus bladder hypertrophy, independent from bladder denervation, causes a shift in the muscarinic receptor subtype mediating bladder contraction from M3 toward M2. PMID- 12763742 TI - IGF-I augments resection-induced mucosal hyperplasia by altering enterocyte kinetics. AB - Our objective was to determine if exogenous insulin-like growth factor-I (IGF-I) augments the adaptive growth response to mid small bowel resection in association with changes in enterocyte kinetics. We determined structural adaptation and concomitant changes in enterocyte proliferation, apoptosis, and migration of the jejunum in growing, parenterally fed rats after mid small bowel resection or small bowel transection, and treatment with IGF-I or vehicle. IGF-I treatment in resected rats significantly increased jejunal mucosal mass by 20% and mucosal concentrations of protein and DNA by 36 and 33%, respectively, above the response to resection alone. The enhancement of resection-induced adaptive growth and cellularity by IGF-I reflected an increase in enterocyte proliferation, an expansion of the proliferative compartment in the crypt, and no further decrease in enterocyte apoptosis or increase in enterocyte migration beyond the effects of resection. The ability of IGF-I to augment the mucosal hyperplasia stimulated by the endogenous response to resection substantiates the role of IGF-I as an intestinal mitogen that promotes tissue regeneration. PMID- 12763743 TI - Inhibition of neuronal nitric oxide synthase by 7-nitroindazole attenuates acute lung injury in an ovine model. AB - Nitric oxide (NO) has been shown to play a major role in acute lung injury (ALI) after smoke inhalation. In the present study, we developed an ovine sepsis model, created by exposing sheep to smoke inhalation followed by instillation of bacteria into the airway, that mimics human sepsis and pneumonia. We hypothesized that the inhibition of neuronal NO synthase (nNOS) might be beneficial in treating ALI associated with this model. Female sheep (n = 26) were surgically prepared for the study and given a tracheostomy. This was followed by insufflation of 48 breaths of cotton smoke (40 degrees C) into the airway of each animal and subsequent instillation of live Pseudomonas aeruginosa [5 x 10(11) colony forming units (CFU)] into each sheep's lung. All sheep were mechanically ventilated using 100% O2. Continuous infusion of 7-nitroindazole (7-NI), an nNOS inhibitor, NG-monomethyl-l-arginine (l-NMMA), a nonspecific NOS inhibitor, or aminoguanidine (AG), an inducible NOS inhibitor, was started 1 h after insult. The administration of 7-NI improved pulmonary gas exchange (PaO2/FiO2; where PaO2 is arterial PO2 and FiO2 is fractional inspired oxygen concentration) and pulmonary shunt fraction and attenuated the increase in lung wet-to-dry weight ratio seen in the nontreated sheep. Histologically, 7-NI prevented airway obstruction. The increase in airway blood flow after injury in the nontreated group was significantly inhibited by 7-NI. The increase in plasma concentration of nitrate and nitrite (NOx) was inhibited by 7-NI as well. Posttreatment with l NMMA improved the pulmonary gas exchange, but AG did not. The results of the present study show that nNOS may be involved in the pathogenesis of ALI after smoke inhalation injury followed by bacterial instillation in the airway. PMID- 12763744 TI - Right ventricular volume measurement by conductance catheter. AB - Continuous ventricular volume measurement by the conductance method assumes a homogeneous electrical field dispersed throughout and contained within the ventricle. Because of dense trabeculation and complex geometry, right ventricular (RV) volume description by this method may be seriously compromised. This study sought to determine the accuracy and limitations of RV volume measurement by conductance, with magnetic resonance (MR) imaging (MRI) used as a reference, in the porcine RV. Anesthetized pigs (n = 5, 45-55 kg) were placed in a 1.5-T magnet, and ECG-gated transverse MR images (5-mm slices) were acquired during the complete cardiac cycle. RV cavity volumes were subsequently determined by Simpson's technique. Animals were then instrumented with an RV conductance catheter and an ultrasonic pulmonary artery flow probe. Conductance catheter signals were recorded using single- and dual-field (SF and DF) excitation, and the saline-dilution technique was used to correct volumes for parallel conductance. The gain factor (alpha) was calculated as the ratio of conductance- to MRI-derived stroke volume (alpha SV). Variation of alpha during the cardiac cycle was computed by comparing RV conductance volumes with 1) MRI volumes at isochronal time points within the cardiac cycle [alpha(t)] and 2) the pulmonary flow integral during ejection. After calibration, the conductance-MRI volume relation was modeled linearly with good correlation [r = 0.96 (SF) and r = 0.94 (DF)], close to the line of identity. Individual conductance-MRI plots displayed a slight curvilinear relation that was concave toward the MRI axis. Consistent with this finding, alpha(t) varied significantly during the cardiac cycle (0.49 and 0.39 by SF for end systole and end diastole, respectively, P = 0.011). DF excitation resulted in improved volume measurement [alpha SV = 0.41 (SF) and 0.96 (DF)], with less variation in alpha(t) (1.0 and 0.92 by DF for end systole and end diastole, respectively, P = 0.66). These results indicate that, with calibration, the conductance method can measure absolute RV volume under steady state conditions. However, the curvilinearity and alpha(t) variation would indicate the potential for nonlinearity when RV volumes are varied over a wider range. PMID- 12763746 TI - Vascular endothelial growth factor mRNA expression and arteriovenous balance in response to prolonged, submaximal exercise in humans. AB - Angiogenesis, the growth of new blood vessels from existing ones, occurs in the skeletal muscle as an adaptive response to exercise that satisfies the increased requirement of this tissue for oxygen delivery and metabolic processes. Of the factors that have been identified to regulate this process, the endothelial cell mitogen vascular endothelial growth factor (VEGF) has been proposed to play a key role. The aim of this study was to measure the skeletal muscle VEGF mRNA content and arteriovenous protein balance across the working leg in response to a single bout of prolonged, submaximal exercise. Seven physically active males completed 3 h of two-legged kicking ergometry. Muscle biopsies were collected from the vastus lateralis muscle from both working legs, and blood samples were collected from one femoral artery and femoral vein before, during, and in recovery from exercise. We show that the exercise stimulus elicited a decrease in VEGF protein arteriovenous balance across the exercising leg (P = 0.007), and a ninefold elevation in skeletal muscle VEGF mRNA expression (P < 0.001). The changes in VEGF protein balance and mRNA content were most pronounced 1 h after the cessation of exercise. In conclusion, these findings demonstrate that submaximal exercise, suitable for humans with low CV fitness, induces a decrease in VEGF arteriovenous balance that is likely to be of clinical significance in promoting angiogenic effects. PMID- 12763745 TI - Cardioprotection through a PKC-dependent decrease in myofilament ATPase. AB - Activation of myocardial kappa-opioid receptor-protein kinase C (PKC) pathways may improve postischemic contractile function through a myofilament reduction in ATP utilization. To test this, we first examined the effects of PKC inhibitors on kappa-opioid receptor-dependent cardioprotection. The kappa-opioid receptor agonist U50,488H (U50) increased postischemic left ventricular developed pressure and reduced postischemic end-diastolic pressure compared with controls. PKC inhibitors abolished the cardioprotective effects of U50. To determine whether kappa-opioid-PKC-dependent decreases in Ca2+-dependent actomyosin Mg2+-ATPase could account for cardioprotection, we subjected hearts to three separate actomyosin ATPase-lowering protocols. We observed that moderate decreases in myofibrillar ATPase were equally cardioprotective as kappa-opioid receptor stimulation. Immunoblot analysis and confocal microscopy revealed a kappa-opioid induced increase in myofilament-associated PKC-epsilon, and myofibrillar Ca2+ independent PKC activity was increased after kappa-opioid stimulation. This PKC myofilament association led to an increase in troponin I and C-protein phosphorylation. Thus we propose PKC-epsilon activation and translocation to the myofilaments causes a decrease in actomyosin ATPase, which contributes to the kappa-opioid receptor-dependent cardioprotective mechanism. PMID- 12763747 TI - Role of dual-site phospholamban phosphorylation in the stunned heart: insights from phospholamban site-specific mutants. AB - Phosphorylation of phospholamban (PLB) at Ser16 (protein kinase A site) and at Thr17 [Ca2+/calmodulin kinase II (CaMKII) site] increases sarcoplasmic reticulum Ca2+ uptake and myocardial contractility and relaxation. In perfused rat hearts submitted to ischemia-reperfusion, we previously showed an ischemia-induced Ser16 phosphorylation that was dependent on beta-adrenergic stimulation and an ischemia and reperfusion-induced Thr17 phosphorylation that was dependent on Ca2+ influx. To elucidate the relationship between these two PLB phosphorylation sites and postischemic mechanical recovery, rat hearts were submitted to ischemia reperfusion in the absence and presence of the CaMKII inhibitor KN-93 (1 microM) or the beta-adrenergic blocker dl-propranolol (1 microM). KN-93 diminished the reperfusion-induced Thr17 phosphorylation and depressed the recovery of contraction and relaxation after ischemia. dl-Propranolol decreased the ischemia induced Ser16 phosphorylation but failed to modify the contractile recovery. To obtain further insights into the functional role of the two PLB phosphorylation sites in postischemic mechanical recovery, transgenic mice expressing wild-type PLB (PLB-WT) or PLB mutants in which either Thr17 or Ser16 were replaced by Ala (PLB-T17A and PLB-S16A, respectively) into the PLB-null background were used. Both PLB mutants showed a lower contractile recovery than PLB-WT. However, this recovery was significantly impaired all along reperfusion in PLB-T17A, whereas it was depressed only at the beginning of reperfusion in PLB-S16A. Moreover, the recovery of relaxation was delayed in PLB-T17A, whereas it did not change in PLB S16A, compared with PLB-WT. These findings indicate that, although both PLB phosphorylation sites are involved in the mechanical recovery after ischemia, Thr17 appears to play a major role. PMID- 12763748 TI - Elevated glucose impairs cAMP-mediated dilation by reducing Kv channel activity in rat small coronary smooth muscle cells. AB - Hyperglycemia impairs endothelium-dependent vasodilation. In this study, we examined the effect of high glucose (HG) on vascular smooth muscle function. Rat small coronary arteries were freshly isolated or incubated for 24 h with normal glucose (NG; 5.5 mmol/l) or HG (23 mmol/l). In freshly isolated arteries, dilation to isoproterenol (Iso) was reduced by 3 mmol/l 4-aminopyridine (4-AP; 44 +/- 10% vs. 77 +/- 4%; P < 0.05) and further reduced by 4-AP + iberiotoxin (IbTX; 100 nmol/l; 17 +/- 2%). Dilation to forskolin was abolished by 4-AP (-3 +/- 17 vs. 73 +/- 9%). cAMP production was similar in NG and HG vessels. Dilations to Iso and forskolin were significantly reduced in HG arteries (Iso, 41 +/- 5% vs. 70 +/- 6%; forskolin, 40 +/- 4% vs. 75 +/- 4%) compared with NG arteries. A similar reduction was also observed to the dilation to papaverine. Endothelial denudation had no effect on Iso-induced dilation. In HG vessels, the reduced 4-AP sensitive component of Iso-induced dilation was greater compared with the IbTX sensitive component. Iso increased whole cell K+ current in NG cells but had little effect in HG cells. Similarly, 4-AP-, but not IbTX-sensitive, K+ currents were reduced in HG cells. These results suggest that HG impairs cAMP-mediated dilation primarily by reducing Kv channel function. We speculate that in addition to the endothelial dysfunction, altered smooth muscle function may also contribute to the reduced coronary vasodilation in diabetes. PMID- 12763749 TI - Basic fibroblast growth factor increases collateral blood flow in spontaneously hypertensive rats. AB - Ischemia-induced angiogenic response is reduced in spontaneously hypertensive rats (SHR). To study whether exogenous basic fibroblast growth factor (bFGF) infusion is effective in expanding collateral circulation in frankly hypertensive SHR, femoral arteries of male SHR (weighing approximately 250 g) were kept intact (nonoccluded control; n = 9) or occluded for 4h(n = 12) or for 16 days with vehicle (n = 14) or bFGF [0.5 (n = 17), 5.0 (n = 13), and 50.0 (n = 14) microg. kg-1. day-1 for 14 days] intraarterially. Maximal collateral-dependent blood flows (BF) to the hindlimbs were determined with 85Sr- and 141Ce-labeled microspheres during running at 20 and 25 m/min (15% grade). Preexercise heart rates (approximately 530 beats/min) and blood pressures (BP; approximately 200 mmHg) were similar across groups except in the high-dose bFGF group, where BP was reduced by approximately 12% (P < 0.05). Femoral artery occlusion for 4 h resulted in approximately 95% reduction of BF in calf muscles [199 +/- 18.7 (nonoccluded group) to 10 +/- 1.0 ml. min-1. 100 g-1; P < 0.001]. BF to calf muscles of the vehicle and low-dose bFGF (0.5 microg. kg-1. day-1) groups increased to 36 +/- 3.2 and 45 +/- 2.0 ml. min-1. 100 g-1, respectively (P < 0.001). bFGF infusion at 5.0 and 50.0 microg. kg-1. day-1 further increased (P < 0.001) BF to calf muscles (62 +/- 4.6 and 62 +/- 2.2 ml. min-1. 100 g-1, respectively). Our results show that bFGF can effectively increase BF in hypertensive rats. The reduced hypertension with high-dose bFGF suggests that a critical signal in arteriogenesis (nitric oxide bioavailability) may be restored. These findings suggest that the dulled endothelial nitric oxide synthase of SHR does not preempt collateral vessel remodeling. PMID- 12763750 TI - Chronic shear induces caveolae formation and alters ERK and Akt responses in endothelial cells. AB - Caveolae are plasmalemmal domains enriched with cholesterol, caveolins, and signaling molecules. Endothelial cells in vivo are continuously exposed to shear conditions, and their caveolae density and location may be different from that of static cultured cells. Here, we show that chronic shear exposure regulates formation and localization of caveolae and caveolin-1 in bovine aortic endothelial cells (BAEC). Chronic exposure (1 or 3 days) of BAEC to laminar shear increased the total number of caveolae by 45-48% above static control. This increase was due to a rise in the luminal caveolae density without changing abluminal caveolae numbers or increasing caveolin-1 mRNA and protein levels. Whereas some caveolin-1 was found in the plasma membrane in static-cultured cells, it was predominantly localized in the Golgi. In contrast, chronic shear exposed cells showed intense caveolin-1 staining in the luminal plasma membrane with minimum Golgi association. The preferential luminal localization of caveolae may play an important role in endothelial mechanosensing. Indeed, we found that chronic shear exposure (preconditioning) altered activation patterns of two well known shear-sensitive signaling molecules (ERK and Akt) in response to a step increase in shear stress. ERK activation was blunted in shear preconditioned cells, whereas the Akt response was accelerated. These results suggest that chronic shear stimulates caveolae formation by translocating caveolin-1 from the Golgi to the luminal plasma membrane and alters cell signaling responses. PMID- 12763751 TI - Preconditioning attenuates ischemia-reperfusion-induced remodeling of Na+-K+ ATPase in hearts. AB - The aim of this study was to determine whether changes in protein content and/or gene expression of Na+-K+-ATPase subunits underlie its decreased enzyme activity during ischemia and reperfusion. We measured protein and mRNA subunit levels in isolated rat hearts subjected to 30 min of ischemia and 30 min of reperfusion (I/R). The effect of ischemic preconditioning (IP), induced by three cycles of ischemia and reperfusion (10 min each), was also assessed on the molecular changes in Na+-K+-ATPase subunit composition due to I/R. I/R reduced the protein levels of the alpha2-, alpha3-, beta1-, and beta2-isoforms by 71%, 85%, 27%, and 65%, respectively, whereas the alpha1-isoform was decreased by <15%. A similar reduction in mRNA levels also occurred for the isoforms of Na+-K+-ATPase. IP attenuated the reduction in protein levels of Na+-K+-ATPase alpha2-, alpha3-, and beta2-isoforms induced by I/R, without affecting the alpha1- and beta1-isoforms. Furthermore, IP prevented the reduction in mRNA levels of Na+-K+-ATPase alpha2-, alpha3-, and beta1-isoforms following I/R. Similar alterations in protein contents and mRNA levels for the Na+/Ca2+ exchanger were seen due to I/R as well as IP. These findings indicate that remodeling of Na+-K+-ATPase may occur because of I/R injury, and this may partly explain the reduction in enzyme activity in ischemic heart disease. Furthermore, IP may produce beneficial effects by attenuating the remodeling of Na+-K+-ATPase and changes in Na+/Ca2+ exchanger in hearts after I/R. PMID- 12763752 TI - Remodeling of cardiac fiber structure after infarction in rats quantified with diffusion tensor MRI. AB - Structural remodeling of myocardium after infarction plays a critical role in functional adaptation. Diffusion tensor magnetic resonance imaging (DTMRI) provides a means for rapid and nondestructive characterization of the three dimensional fiber architecture of cardiac tissues. In this study, microscopic structural changes caused by MI were evaluated in Fischer 344 rats 4 wk after infarct surgery. DTMRI studies were performed on 15 excised, formalin-fixed rat hearts of both infarct (left anterior descending coronary artery occlusion, n = 8) and control (sham, n = 7) rats. Infarct myocardium exhibited increased water diffusivity (41% increase in trace values) and decreased diffusion anisotropy (37% decrease in relative anisotropy index). The reduced diffusion anisotropy correlated negatively with microscopic fiber disarray determined by histological analysis (R = 0.81). Transmural courses of fiber orientation angles in infarct zones were similar to those of normal myocardium. However, regional angular deviation of the diffusion tensor increased significantly in the infarct myocardium and correlated strongly with microscopic fiber disarray (R = 0.86). These results suggest that DTMRI may provide a valuable tool for defining structural remodeling in diseased myocardium at the cellular and tissue level. PMID- 12763753 TI - Nucleotide coronary vasodilation in guinea pig hearts. AB - The role of P1 receptors and P2Y1 receptors in coronary vasodilator responses to adenine nucleotides was examined in the isolated guinea pig heart. Bolus arterial injections of nucleotides were made in hearts perfused at constant pressure. Peak increase in flow was measured before and after addition of purinoceptor antagonists. Both the P1 receptor antagonist 8-(p-sulfophenyl)theophylline and adenosine deaminase inhibited adenosine vasodilation. AMP-induced vasodilation was inhibited by P1 receptor blockade but not by adenosine deaminase or by the selective P2Y1 antagonist N6-methyl-2'-deoxyadenosine 3',5'-bisphosphate (MRS 2179). ADP-induced vasodilation was moderately inhibited by P1 receptor blockade and greatly inhibited by combined P1 and P2Y1 blockade. ATP-induced vasodilation was antagonized by P1 blockade but not by adenosine deaminase. Addition of P2Y1 blockade to P1 blockade shifted the ATP dose-response curve further rightward. It is concluded that in this preparation ATP-induced vasodilation results primarily from AMP stimulation of P1 receptors, with a smaller component from ATP or ADP acting on P2Y1 receptors. ADP-induced vasodilation is largely due to P2Y1 receptors, with a smaller contribution by AMP or adenosine acting via P1 receptors. AMP responses are mediated solely by P1 receptors. Adenosine contributes very little to vasodilation resulting from bolus intracoronary injections of ATP, ADP, or AMP. PMID- 12763754 TI - Angiotensin II modulates nitric oxide-induced cardiac fibroblast apoptosis by activation of AKT/PKB. AB - Previously we found that interleukin-1beta (IL-1beta)-activated inducible nitric oxide (NO) synthase (iNOS) expression and that NO production can trigger cardiac fibroblast (CFb) apoptosis. Here, we provide evidence that angiotensin II (ANG II) significantly attenuated IL-1beta-induced iNOS expression and NO production in CFbs while simultaneously decreasing apoptotic frequency. The anti-apoptotic effect of ANG II was abolished when cells were pretreated with the specific ANG II type 1 receptor (AT1) antagonist losartan, but not by the AT2 antagonist DP 123319. Furthermore, ANG II also protected CFbs from apoptosis induced by the NO donor diethylenetriamine NONOate and this effect was associated with phosphorylation of Akt/protein kinase B at Ser473. The effects of ANG II on Akt phosphorylation and NO donor-induced CFb apoptosis were abrogated when cells were preincubated with the specific phosphatidylinositol 3-kinase inhibitors wortmannin or LY-294002. These data demonstrate that ANG II protection of CFbs from IL-1beta-induced apoptosis is associated with downregulation of iNOS expression and requires an intact phosphatidylinositol 3-kinase-Akt survival signal pathway. The findings suggest that ANG II and NO may play a role in regulating the cell population size by their countervailing influences on cardiac fibroblast viability. PMID- 12763755 TI - Proliferation, maturation and germination of Castanea sativa Mill. Somatic embryos originated from leaf explants. AB - Experiments were performed to determine the influence of proliferation medium on the maintenance of embryogenic competence and on repetitive embryogenesis in Castanea sativa Mill. somatic embryos derived from leaf explants. Somatic embryo proliferation was carried out by both direct secondary embryogenesis and by the culture of nodular callus tissue originated from cotyledons of somatic embryos. Both systems led to the production of cotyledonary somatic embryos on Murashige and Skoog proliferation medium supplemented with 0.1 mg l-1 benzyladenine and 0.1 mg l-1 naphthaleneacetic acid. Carbon source and concentration had a marked influence on maturation and subsequent germination ability of chestnut somatic embryos. Plantlet conversion was achieved in embryos matured on media with 6 % sucrose, and on 3 or 6 % maltose, whereas mean shoot length, root length and leaf number of produced plants were not significantly affected by these maturation media. Overall, the best results were obtained with 3 % maltose-matured somatic embryos, giving rise to 6 % plant recovery in addition to 33 % of embryos exhibiting only shoot development. The application of a 2-month cold treatment at 4 degrees C to somatic embryos matured on medium with 3 % maltose was necessary for achieving plant conversion, while partial desiccation did not appear to influence this response. A total of 39 % of embryos eventually produced plants either through conversion to plantlets or indirectly through rooting of shoots. Shoots formed by somatic embryos could be excised, multiplied and rooted following the micropropagation procedures previously developed for chestnut. PMID- 12763756 TI - Girdling affects carbohydrate-related gene expression in leaves, bark and roots of alternate-bearing citrus trees. AB - Effects of girdling on carbohydrate status and carbohydrate-related gene expression in citrus trees were investigated. Alternate-bearing 'Murcott' (a Citrus reticulata hybrid of unknown origin) trees were girdled during autumn (25 Sep. 2001) and examined 10 weeks later. Girdling brought about carbohydrate (soluble sugar and starch) accumulation in leaves and shoot bark above the girdle, in trees during their fruitless, 'off' year. Trees during their heavy fruit load, 'on' year did not accumulate carbohydrates above the girdle due to the high demand for carbohydrates by the developing fruit. Girdling caused a strong decline in soluble sugar and starch concentrations in organs below the girdle (roots), in both 'on' and 'off' trees. Expression of STPH-L and STPH-H (two isoforms of starch phosphorylase), Agps (ADP-glucose pyrophosphorylase, small subunit), AATP (plastidic ADP/ATP transporter), PGM-C (phosphoglucomutase) and CitSuS1 (sucrose synthase), all of which are associated with starch accumulation, was studied. It was found that gene expression is related to starch accumulation in all 'off' tree organs. RNA levels of all the genes examined were high in leaves and bark that accumulated high concentrations of starch, and low in roots with declining starch concentrations. It may be hypothesized that changes in specific sugars signal the up- and down-regulation of genes involved in starch synthesis. PMID- 12763757 TI - Fracture of plant tissues and walls as visualized by environmental scanning electron microscopy. AB - The environmental scanning electron microscope (ESEM) provides a highly relevant and controllable environment in which to study hydrated systems without the artefacts of other highly prepared specimens. The instrument facilitates control of turgor through hydration using different chamber vapour pressures. Deformation of a simple plant tissue-upper epidermal layers in Allium cepa (onion)-was observed at the scale of the two principal failure mechanisms: cell breakage; and cell separation induced by treatment with a chelating agent. Cell rupture and release of contents occurred at cellular junctions ahead of an imposed growing notch, indicating that disruption of cells occurred remotely from the creation of a new surface. Cells that separated usually maintained their turgor and the separation process took place through progressive failure of middle lamellar material seen as strands between separating cells. These mechanisms were compared with the rupture of excised Chara corallina walls that occurred by formation and breakage of strands between separating wall layers. This study provides in situ visual characterization of wall rupture and cell separation at the microscopic level in hydrated plant material. PMID- 12763758 TI - Palaeo-adaptive properties of the xylem of Metasequoia: mechanical/hydraulic compromises. AB - The xylem of Metasequoia glyptostroboides Hu et Cheng is characterized by very low density (average specific gravity = 0.27) and tracheids with relatively large dimensions (length and diameter). The microfibril angle in the S2 layer of tracheid walls is large, even in outer rings, suggesting a cambial response to compressive rather than tensile stresses. In some cases, this compressive stress is converted to irreversible strain (plastic deformation), as evidenced by cell wall corrugations. The heartwood is moderately decay resistant, helping to prevent Brazier buckling. These xylem properties are referenced to the measured bending properties of modulus of rupture and modulus of elasticity, and compared with other low-to-moderate density conifers. The design strategy for Metasequoia is to produce a mechanically weak but hydraulically efficient xylem that permits rapid height growth and crown development to capture and dominate a wet site environment. The adaptability of these features to a high-latitude Eocene palaeoenvironment is discussed. PMID- 12763759 TI - Seasonal variation in respiration of 1-year-old shoots of scots pine exposed to elevated carbon dioxide and temperature for 4 years. AB - Sixteen 20-year-old Scots pine (Pinus sylvestris L.) trees growing in the field were enclosed for 4 years in environment-controlled chambers that maintained: (1) ambient conditions (CON); (2) elevated atmospheric CO2 concentration (ambient + 350 micro mol mol-1; EC); (3) elevated temperature (ambient +2-6 degrees C; ET); or (4) elevated CO2 and elevated temperature (ECT). The dark respiration rates of 1-year-old shoots, from which needles had been partly removed, were measured over the growing season in the fourth year. In all treatments, the temperature coefficient of respiration, Q10, changed with season, being smaller during the growing season than at other times. Respiration rate varied diurnally and seasonally with temperature, being highest around mid-summer and declining gradually thereafter. When measurements were made at the temperature of the chamber, respiration rates were reduced by the EC treatment relative to CON, but were increased by ET and ECT treatments. However, respiration rates at a reference temperature of 15 degrees C were reduced by ET and ECT treatments, reflecting a decreased capacity for respiration at warmer temperatures (negative acclimation). The interaction between season and treatment was not significant. Growth respiration did not differ between treatments, but maintenance respiration did, and the differences in mean daily respiration rate between the treatments were attributable to the maintenance component. We conclude that maintenance respiration should be considered when modelling respiratory responses to elevated CO2 and elevated temperature, and that increased atmospheric temperature is more important than increasing CO2 when assessing the carbon budget of pine forests under conditions of climate change. PMID- 12763760 TI - Efflux and atherosclerosis: the clinical and biochemical impact of variations in the ABCA1 gene. AB - Approximately 50 mutations and many single nucleotide polymorphisms have been described in the ABCA1 gene, with mutations leading to Tangier disease and familial hypoalphalipoproteinemia. Homozygotes and heterozygotes for mutations in ABCA1 display a wide range of phenotypes. Identification of ABCA1 as the molecular defect in these diseases has allowed for ascertainment based on genetic status and determination of genotype-phenotype correlations and has permitted us to identify mutations conferring a range of severity of cellular, biochemical, and clinical phenotypes. In this study we review how genetic variation at the ABCA1 locus affects its role in the maintenance of lipid homeostasis and the natural progression of atherosclerosis. PMID- 12763761 TI - The p17 cleaved form of caspase-3 is present within viable macrophages in vitro and in atherosclerotic plaque. AB - OBJECTIVE: In vitro studies of macrophage death in response to oxidized LDL (oxLDL) were undertaken as a model for the formation of the necrotic core of atherosclerotic plaque. METHODS AND RESULTS: Thioglycollate-elicited mouse peritoneal macrophages avidly incorporated both oxLDL and acetylated LDL (acLDL) to become foam cells. oxLDL-treated macrophages, but not acLDL-treated macrophages, showed nearly 100% death, with characteristics consistent with apoptosis, including cell surface phosphatidylserine exposure, intracellular caspase-3 activity, cleavage of caspase-3 substrates, and DNA fragmentation, as shown by TUNEL assay. The activated form of caspase-3 (p17 cleaved form) was present in attached, viable macrophages before exposure to oxLDL. This p17 form was also found in apparently viable as well as in TUNEL-positive cells within atherosclerotic lesions of chow-fed apolipoprotein E-deficient (ApoE-/-) mice. The amount of p17 caspase-3 was reduced by in vitro blockade of FasL with an FasL blocking antibody and was absent in macrophages from lpr/lpr mice, which lack functional Fas. Moreover, lpr/lpr macrophages resisted oxLDL cytotoxicity. CONCLUSIONS: The naturally occurring Fas-FasL induction of caspase-3 cleavage after macrophage attachment may represent an important physiologic mechanism that primes for cytotoxicity by oxLDL and possibly, other death-inducing molecules. PMID- 12763762 TI - Periodontal disease is associated with brachial artery endothelial dysfunction and systemic inflammation. AB - OBJECTIVE: The purpose of this study was to determine whether periodontal disease is associated with endothelial dysfunction and systemic inflammation. Epidemiological studies suggest that severe periodontal disease is associated with increased cardiovascular disease risk, but the mechanisms remain unknown. METHODS AND RESULTS: We assessed flow-mediated dilation and nitroglycerin mediated dilation of the brachial artery using vascular ultrasound in 26 subjects with advanced periodontal disease and 29 control subjects. The groups were matched for age and sex, and patients with hypercholesterolemia, diabetes mellitus, hypertension, and history of cigarette smoking were excluded. We also examined serum levels of C-reactive protein using an established high-sensitivity method. Subjects with advanced periodontal disease had lower flow-mediated dilation compared with control patients (7.8+/-4.6% versus 11.7+/-5.3%, P=0.005). Nitroglycerin-mediated dilation was equivalent in the two groups. Subjects with advanced periodontitis exhibited higher serum levels of high-sensitivity C reactive protein compared with healthy controls patients (2.3+/-2.3 versus 1.0+/ 1.0 mg/L, P=0.03). CONCLUSIONS: Subjects with advanced periodontal disease exhibit endothelial dysfunction and evidence of systemic inflammation, possibly placing them at increased risk for cardiovascular disease. PMID- 12763763 TI - Angiotensin II inhibits endothelial cell motility through an AT1-dependent oxidant-sensitive decrement of nitric oxide availability. AB - OBJECTIVE: The migratory capability of vascular endothelial cells plays a pivotal role in the maintenance of vessel wall integrity and is stimulated by nitric oxide (NO). Angiotensin II increases NAD(P)H oxidase activity in endothelial cells, thereby promoting reactive oxygen species (ROS) generation. Because ROS can both reduce NO synthase activity and increase NO breakdown, thus impairing NO availability in endothelial cells, we evaluated the effect of angiotensin II on human vascular endothelial cell (HUVEC) motility. METHODS AND RESULTS: Angiotensin II dose- and time-dependently reduced HUVEC migration. Besides inhibiting HUVEC motility, angiotensin II altered intracellular glutathione redox status. The generation of ROS by cultured HUVECs was significantly increased by angiotensin II. Furthermore, angiotensin II reduced NO metabolite concentrations in culture media. The angiotensin II type 1 receptor antagonist candesartan cilexetil attenuated the inhibitory action exerted by angiotensin II on HUVEC motility, reversed the angiotensin II-induced increase in intracellular oxidative stress, and restored NO availability. Similar effects were exerted by the flavonoid inhibitor diphenylene iodinium and the antioxidant agent N-acetyl-L cysteine. CONCLUSIONS: All together, our data demonstrate that angiotensin II inhibits HUVEC motility by reducing NO availability. Such reduction is due to an angiotensin II type 1 receptor-dependent increment in intracellular ROS generation. PMID- 12763766 TI - Proceedings of the 10th International Conference on Na,K-ATPase and Related Cation Pumps. August 8-14, 2002, Elsinore, Denmark. PMID- 12763764 TI - Electron spin resonance detection of hydrogen peroxide as an endothelium-derived hyperpolarizing factor in porcine coronary microvessels. AB - OBJECTIVE: Endothelium-derived hyperpolarizing factor (EDHF) plays an important role in modulating vascular tone, especially in microvessels, although its nature has yet to be elucidated. This study was designed to examine whether hydrogen peroxide (H2O2) is an EDHF in porcine coronary microvessels with use of an electron spin resonance (ESR) method to directly detect H2O2 production from the endothelium. METHODS AND RESULTS: Isometric tension and membrane-potential recordings demonstrated that bradykinin and substance P caused EDHF-mediated relaxations and hyperpolarizations of porcine coronary microvessels in the presence of indomethacin and Nomega-nitro-L-arginine. The contribution of H2O2 to the EDHF-mediated responses was demonstrated by the inhibitory effect of catalase and by the relaxing and hyperpolarizing effects of exogenous H2O2. Endothelial production of H2O2 was quantified in bradykinin- or substance P-stimulated intact blood vessels by ESR spectroscopy. Tiron, a superoxide scavenger that facilitates H2O2 formation, enhanced bradykinin-induced production of H2O2, as well as the EDHF-mediated relaxations and hyperpolarizations. By contrast, cytochrome P-450 inhibitors (sulfaphenazole or 17-octadecynoic acid) or a gap junction inhibitor (18alpha-glycyrrhetinic acid) failed to inhibit the EDHF-mediated relaxations. Involvement of endothelium-derived K+ was not evident in experiments with ouabain plus Ba2+ or exogenous K+. CONCLUSIONS: These results provide ESR evidence that H2O2 is an EDHF in porcine coronary microvessels. PMID- 12763767 TI - Crystal structures of Ca2+-ATPase in various physiological states. AB - The structures of the Ca(2+)-ATPase (SERCA1a) in different physiological states were determined by X-ray crystallography. Detailed comparison of the structures in the Ca(2+)-bound form and unbound (but thapsigargin bound) form reveals that very large rearrangements of the transmembrane helices take place accompanying Ca(2+) dissociation and binding and that they are mechanically linked with equally large movements of the cytoplasmic domains. The meaning of the rearrangement of the transmembrane helices becomes apparent by homology modeling of the Na(+)K(+)-ATPase. PMID- 12763768 TI - Renal Na,K-ATPase structure from cryo-electron microscopy of two-dimensional crystals. AB - The molecular structure of Na,K-ATPase was determined by electron crystallography from two-dimensional crystals induced in purified membranes isolated from the outer medulla of pig kidney. The P2 type unit cell contains two protomers in the E(2) conformation, each of them with a size of 65 x 75 x 150 A(3). The alpha, beta, and gamma subunits in the membrane crystals were demonstrated in the crystals with Western blotting and related to distinct domains in the density map. The alpha subunit corresponds to most of the density in the transmembrane region as well as to the large hydrophilic headpiece on the cytoplasmic side of the membrane. The headpiece is divided into three separated domains. One of these gives rise to an elongated projection onto the membrane plane, while the putative nucleotide binding and phosphorylation domains form compact densities in the rest of the cytoplasmic part of the structure. Density on the extracellular face corresponds to the protein part of the beta subunit. Ten helices from the catalytic a subunit correspond to two groups of distinct densities in the transmembrane region. The structure of the lipid bilayer spanning part also suggests positions for the transmembrane helices from the beta and gamma subunits. The overall structure of the alpha subunit of Na,K-ATPase as determined here by cryo-electron microscopy is similar to the X-ray structure of Ca-ATPase. However, conformational changes between the E(1) and E(2) forms are suggested by different relative positions of cytoplasmic domains. PMID- 12763769 TI - Projection map of covalently phosphorylated Ca-ATPase from tubular crystals. PMID- 12763770 TI - How does conformation change ouabain binding from rejection (E1) to acceptance (E2)? PMID- 12763771 TI - Transmission of E1-E2 structural changes in response to Na+ or K+ binding in Na,K ATPase. AB - The extensive E(1)-E(2) conformational changes in response to Na(+) or K(+) binding in the absence of other ligands must be driven by motion of the side chains contributing to cation coordination, but the differences in structure of Na(+) and K(+) sites have not been resolved. The recent high resolution structure model of the E(2) conformation of Ca-ATPase offers the first opportunity to examine and model the changes accompanying the adjustment of the cation sites from an E(1) form with specificity for Na(+) to an E(2) form with specificity for K(+). The model of the E(2) form provides a remarkable fit to the data of direct Tl(+) or K(+) binding after site-directed mutagenesis of residues Asp804 and Asp808 in M6, Glu 779, Gln776, and Ser775 in M5, and Glu327 in M4. Cytoplasmic domain movements during E(1) <--> E(2) conformational transition can be monitored by proteolytic cleavage. Protection of the chymotrypsin-sensitive bond at Leu266 in L2/3 and rotation of the A domain is more complete in the E(2)Mg-vanadate ouabain complex than in the E(2)[2K] form. PMID- 12763772 TI - Conformational dynamics of Na+/K+- and H+/K+-ATPase probed by voltage clamp fluorometry. AB - We used the method of site-directed fluorescence labeling in combination with voltage-clamp fluorometry for time-resolved recording of localized conformational transitions of the Na(+)/K(+)- and H(+)/K(+)-ATPase. Therefore, single cysteine mutations were introduced into the extracellular TM5-TM6 loop of the sheep Na(+)/K(+)-ATPase alpha(1)-subunit devoid of other extracellular cysteines. Upon expression in Xenopus oocytes and covalent attachment of tetramethylrhodamine maleimide (TMRM) as a reporter fluorophore, Cys-mutant N790C showed large fluorescence changes of up to 5% in response to extracellular K(+) that were completely abolished by ouabain. When voltage jumps were applied under Na(+)/Na(+)-exchange conditions, we observed fluorescence changes that paralleled the transient currents originating from the E(1)P<-->E(2)P transition. These fluorescence changes were also completely inhibited by ouabain, as were the voltage jump-induced transient currents. Transient fluorescence changes could also be measured as a function of increasing K(+) concentrations, that is, under turnover conditions. As a result, the distribution between E(1) and E(2) states can be determined at any time and membrane potential. Very similar fluorescence signals were obtained for rat gastric H(+)/K(+)-ATPase upon expression in oocytes, when a single cysteine was introduced at a position homologous to N790 in Na(+)/K(+)-ATPase for attachment of the fluorophore. As to the high sequence similarity among P-type ATPases within the TM5 helix and the TM5-TM6 loop region, our results enable new means of kinetic investigation for these pumps under physiological conditions in living cells. PMID- 12763773 TI - Investigating the energy transduction mechanism of P-type ATPases with Fe2+ catalyzed oxidative cleavage. AB - This paper discusses specific oxidative cleavage of renal Na(+),K(+)-ATPase and gastric H(+),K(+)-ATPase, catalyzed by bound Fe(2+) or the complex ATP-Fe(2+), and its implication for the energy transduction mechanism of P-type ATPases. The cleavage technique provides information on the spatial organization of the proteins in different conformations and, since ATP-Fe(2+) substitutes for ATP Mg(2+) in activating ATPase activity, on Mg(2+)-ligating residues in different conformations. The experiments predict the existence of large movements of N, P, and A cytoplasmic domains accompanying E(1)-E(2) and E(1) P-E(2)P conformational transitions-open in E(1) conformations and closed in E(2) conformations. These features fit well with the Ca(2+)-ATPase crystal structures in E(1) or E(2) conformations and also provide evidence on ATP and Mg(2+) binding sites that is not available from the structures. By combining information from cleavage experiments with molecular modeling, based on the Ca(2+)-ATPase structure, features such as an N to P domain interaction in an E(1). ATP-Mg(2+) conformation can be inferred. The organization of the N, P, and A domains and the ATP and Mg(2+) binding sites in the different conformational states appears to be essentially similar for Na(+),K(+)-ATPase, H(+),K(+)-ATPase, and Ca(2+)-ATPase. The oxidative cleavage technique may be a valuable tool to investigate long-range interactions that transduce the free energy of hydrolysis of ATP to active cation movements in P-type ATPases. PMID- 12763774 TI - Importance of transmembrane segment M3 of Na+,K+-ATPase for control of conformational changes and the cytoplasmic entry pathway for Na. AB - A series of mutations were introduced into the sequence Glu(282)-Ile-Glu-His-Phe Ile-His(288) of the NH(2)-terminal part of M3 of the rat kidney Na(+),K(+) ATPase, and the resulting mutant pumps were analyzed functionally. Several of the mutations affected the conformational transitions between E(1) and E(2) forms of dephospho- and phosphoenzyme. Mutations to Glu(282) and Phe(286) affected the E(1)-E(2) and E(1)P-E(2)P equilibria in parallel, indicating a role for these two residues in both conformational changes. Mutation to His(285) preferentially affected the E(1)P-E(2)P equilibrium, and mutation to Ile(283) affected only the E(1)-E(2) equilibrium of the dephosphoenzyme, demonstrating that the conformational changes of M3 in the phospho- and dephospho-forms are not identical. Several of the mutants showed a reduced apparent affinity for Na(+), pointing to an important role of the region in optimizing the Na(+) binding properties of the enzyme. It is possible that this part of M3 is closely associated with an entry pathway through which the Na(+) ions pass from the cytoplasmic surface to reach the cation-binding pocket. Some of the mutants also displayed an increased Na(+)-ATPase activity, and a good correlation was observed between the turnover rate for Na(+)-ATPase activity and the rate of dephosphorylation in the absence of K(+), indicating an increased ability of Na(+) to activate dephosphorylation of E(2)P by binding in place of K(+) at the extracellularly facing sites. Thus, M3 also seems to be a part of the signaling pathway between the external cation sites and the catalytic site. PMID- 12763775 TI - Insights into the structural basis for modulation of E1<-->E2 transitions by cytoplasmic domains of the Na,K-ATPase alpha subunit. PMID- 12763776 TI - Characterization of Ca2+ ATPase residues involved in substrate and cation binding. AB - The role of amino acid residues involved in substrate and cation binding was investigated in complementary experiments on Fe(2+)-catalyzed oxidation and cleavage, limited digestion with proteinase K, and mutational analysis. Cleavage at Ser346 was produced by Fe(2+) in the presence of substrate (ATP or AMP-PNP) and Ca(2+), and was attributed to Fe(2+) bound to a Mg(2+) site near Ser346 and neighboring Glu696. Ca(2+)- and ATP-dependent oxidation of the Thr441 side chain was also observed and attributed to Fe(2+) substituting for Mg(2+) in the Mg(2+) ATP complex bound to the N domain. Mutation of Arg560 or Glu439 within the N domain interfered with nucleotide-dependent ATPase resistance to digestion with proteinase K. Furthermore, mutation of Lys352, Lys684, Thr353, Asp703, or Asp707 within the P domain produced similar interference, consistent with a role of these residues in substrate stabilization at the catalytic site. In a third group of experiments, equilibrium isotherms were obtained with Asn796Ala and Glu309Gln mutants, demonstrating non-cooperative binding of one Ca(2+) per ATPase, as opposed to cooperative binding of two Ca(2+) by WT enzyme. No high-affinity binding by Asp800Asn, Glu771Gln, and Thr799Ala mutants was detected. It was also demonstrated that the conformational transitions involved in enzyme activation and interconversion of Ca(2+) binding and phosphorylation energy, are triggered by Ca(2+) binding to site II and stabilization of Glu309 (M4) and N796 (M6). PMID- 12763777 TI - Mutagenesis of residues involved in control of the Ca2+ entry pathway and conformational changes associated with Ca2+ binding in the SR Ca2+-ATPase. AB - Rapid kinetic measurements were used to study the rate of Ca(2+) dissociation from the high-affinity Ca(2+) sites of the dephosphoenzyme (i.e., from the E(1)Ca(2) form toward the cytoplasmic side) as well as the rate of Ca(2+) binding with associated conformational changes (E(2) --> E(1)Ca(2) transition) in the wild type and mutants of the sarcoplasmic reticulum Ca(2+)-ATPase expressed in mammalian cells. Cluster mutations as well as single mutations in transmembrane segment M3 resulted in conspicuous effects on the rate of Ca(2+) migration. Furthermore, mutation of Asp(59) in transmembrane segment M1 to arginine exerted a profound effect on Ca(2+) interaction. The data demonstrate an important role for M3 residues in control of the Ca(2+) entry pathway and provide functional evidence in support of a close relationship between this pathway and the water accessible channel leading between transmembrane segments M1 and M3 in the thapsigargin stabilized E(2) structure. In addition, rapid kinetic measurements demonstrated that the hydrogen bond network involving Asp(813) of loop L6-7 and Lys(758) of M5 is important for the E(2) --> E(1)Ca(2) transition. PMID- 12763778 TI - Proteolytic studies on the transduction mechanism of sarcoplasmic reticulum Ca2+ ATPase: common features with other P-type ATPases. AB - After proteinase K-induced excision of five amino acid residues in the semiconserved polypeptide chain linking the end of the A domain with the S3/M3 transmembrane segment we find that Ca(2+) transport is blocked while partial reactions like Ca(2+) binding, ATP phosphorylation, and Ca(2+)-occlusion are left intact. However, formation of the so-called E2P state (either from the phosphorylated species formed in the presence of ATP and Ca(2+) or from the Ca(2+)-depleted unphosphorylated species) is blocked. We conclude that the proteinase K-treated ATPase, while maintaining many of the partial reactions, is incapable of energy transduction because of the absence of an E2P state with Ca(2+) binding sites exposed to the intravesicular space. Sequence comparisons and mutagenesis data point to an important role in energy transduction of P-type ATPases of a conserved motif located at the end of the A domain. PMID- 12763779 TI - Involvement of the cytoplasmic loop L6-7 in the entry mechanism for transport of Ca2+ through the sarcoplasmic reticulum Ca2+-ATPase. AB - We have found that despite a markedly low calcium affinity the D813A/D818A mutant is capable, after complexation with Cr.ATP, of occluding Ca(2+) to the same extent (1-2 Ca(2+) per ATPase monomer) as wild- type ATPase. The inherent ability of the synthetic L6-7 loop peptide to bind Ca(2+) was demonstrated with murexide and mass spectrometry. NMR analysis indicated the formation of specific 1:1 cation complexes of the peptide with calcium and lanthanum with coordination by all three aspartate residues D813/D815/D818 that resulted in an altered conformation of the peptide chain. Overall our observations suggest that, in addition to mediating contact between the intramembranous Ca(2+) binding sites and the cytosolic phosphorylation site as previously suggested, the L6-7 loop, in a preceding step, participates in the formation of an entrance port important for lodging Ca(2+) at a high-affinity binding site inside the membrane. PMID- 12763780 TI - Site-directed mutagenesis of amino acids in the cytoplasmic loop 6/7 of Na,K ATPase. AB - The loop between transmembrane helices 6 and 7 (L6/7) of P-type ATPases has been suggested to be important for the functional linkage of ion binding and enzyme phosphorylation or to be a site of initial cation binding. To investigate the role of L6/7 in Na,K-ATPase, alanine substitutions were made for charged and conserved residues in L6/7 of the human alpha1 subunit and the proteins were expressed in yeast for analysis. All mutants except the triple mutant E825A/E828A/D830A bound ouabain. Although the equilibrium dissociation constant for ouabain binding by most mutants was similar to the wild-type value, the K(d) of R837A for ouabain binding was approximately 15-fold higher than the wild-type K(d). (18)O exchange measurements indicated that the apparent affinity of this mutant for Pi was reduced about 3-fold. The concentration dependence of KCl inhibition of ouabain binding or of NaCl inhibition of ouabain binding revealed 2 4-fold changes in the apparent affinity for cations in the E825A, E828A, and R837A mutants. The E825A and E828A mutants lost the ability to bind ouabain after extraction with 0.1% SDS or after brief heating, indicating that these mutations affected the stability of the enzyme. The ATPase activity of the other mutants was measured after extraction of crude yeast membranes with 0.1% SDS. For all mutants except R834A, R837A, and R848A, the activity was at least 50% of wild type activity. PMID- 12763781 TI - ATP binding residues of sarcoplasmic reticulum Ca2+-ATPase. AB - ATP-binding residues in the N and P domains of sarcoplasmic reticulum Ca-ATPase have been investigated using mutagenesis in combination with a binding assay based on the photolabeling of Lys(492) with [g-(32)P] 2',3'-O-(2,4,6 trinitrophenyl)-8-azido-ATP and competition with nucleotide. In the N domain, mutations to several residues in conserved motifs, (438)GEATE, (487)FSRDRK, (515)KGAPE, and (560)RCLALA produce nucleotide-binding defects. Key residues include Thr(441), Glu(442), Phe(487), Arg(489), Lys(492), Lys(515), Arg(560), and Leu(562). In the absence of Mg(2+), Arg(489), Lys(492), and Arg(560) are most important, whereas in its presence Thr(441) and Glu(442) also play a crucial role. In the P domain, Asp(351) is striking for its strong electrostatic repulsion of the gamma-phosphate, especially in the presence of Mg(2+). Lys(352) is a key residue, and Asp(627) and Lys(684) must come close to the nucleotide. Thr(353), Asn(359), Asp(601), and Asp(703) interact only in the presence of Mg(2+). Asn(706) and Asp(707) are unimportant for nucleotide binding. The results identify several ATP binding residues in the N and P domains and suggest that Mg(2+) changes the nucleotide/protein interaction in both. Models of bound ATP and MgATP are presented. PMID- 12763782 TI - Molecular modeling of SCH28080 binding to the gastric H,K-ATPase and MgATP interactions with SERCA- and Na,K-ATPases. AB - We have used homology molecular modeling based on the srCaATPase E(2) conformation, pdb1kju, to predict side chains involved in docking the K(+) competitive inhibitor, SCH28080, to the H,K-ATPase. A model for SCH28080 binding between residues L809 and A335 in the same space utilized by omeprazole is proposed. We also describe modeling MgATP binding to the E(1) structure of the srATPase, pdb1eul, as a paradigm for the Na,K- and H,K-ATPases. The resulting model, E(1).MgATP, visualizes a conformation not yet available by crystallization and successfully predicts a range of published results, including backbone cleavages near V440 (N domain) and V712 (P domain) mediated by FeATP in the Na,K ATPase. A separate model for MgATP docked to E(2) (pdb1kju) shows that access of the gamma phosphate to D351 is blocked by the A domain. The E(2). MgATP model explains FeATP-mediated cleavages of the Na,K-ATPase near V440 and E214 (A domain) and homologous results in the H,K-ATPase. PMID- 12763783 TI - Inhibition kinetics of the gastric H,K-ATPase by K-competitive inhibitor SCH28080 as a tool for investigating the luminal ion pathway. AB - The gastric H,K-ATPase and the Na,K-ATPase both are stimulated by luminal K(+), but differ in sensitivity to K(+)-competitive inhibitors (ouabain and SCH28080), which implies a difference in structure near the luminal ion pathways in these two pumps. Knowledge of the amino acids in the H,K-ATPase that affect the mode of inhibition by SCH28080 and inhibitor affinity should provide insight into the regions of the membrane domain influencing the inhibitor selectivity and the luminal route to the ion transport site. Mutational scans in M4, 5, 6, and 8 have shown that amino acid residues affecting ion affinity (E343, K791, E795, E820, D824, E936) with either no or a lesser effect on the inhibitor affinity are located in the middle of the membrane domain. The residues significantly reducing inhibitor affinity, but not ion affinity (L809, P810, L811, T813, I816, Y925, T929), are located in the exoplasmic 5-6 loop and the luminal ends of M6 and M8. This suggests that the binding domain for SCH28080 contains the surface between L809 in the 5-6 loop and C813 at the luminal end of M6, approximately two helical turns out from the ion binding region, where it blocks an ion access pathway. The mutations that change inhibitor kinetics are on the opposing faces of M6 and M8 and apparently modify the normal ion pathway or, perhaps, create an alternate ion pathway. PMID- 12763784 TI - Ion occlusion/deocclusion partial reactions in individual palytoxin-modified Na/K pumps. AB - In P-type ion-motive ATPases, transported ions approach their binding sites from one membrane surface, become buried deep within "occluded" conformations in which the sites are inaccessible from either membrane side, and are then deoccluded and released to the opposite membrane surface. This describes an alternating-gate transport mechanism, in which the pump acts like an ion channel with two gates that open and close alternately. The occluded states ensure that one gate closes before the other can open, thus preventing the large electrodiffusive ion fluxes that would otherwise quickly undo the pump's electrochemical work. High resolution crystal structures of two conformations of the SERCA (sarcoplasmic and endoplasmic reticulum Ca(2+)) P-type ATPase, together with mutagenesis results and analyses of structural models based on homology, have begun to provide a picture of the ion coordination sites in related P-type ATPases, including the Na/K pump. However, in no P-type ATPase are the structures and mechanisms of the gates known. The marine toxin, palytoxin (PTX), is known to bind to the Na/K pump and elicit a nonselective cation leak pathway, possibly by disrupting the strict coupling between the pump's inner and outer gates, allowing them to both be open. We recently found that ion flow through PTX-modified Na/K pump-channels appears to be modulated by two gates that can be regulated by the pump's physiological ligands in a manner suggesting that gating reflects underlying ion occlusion/deocclusion partial reactions. We review that work here and provide evidence that the pore of the PTX-induced pump-channel has a diameter > 6 A. PMID- 12763785 TI - Cation stoichiometry and cation pathway in the Na,K-ATPase and nongastric H,K ATPase. AB - The mechanism of cation translocation by the Na,K-ATPase was investigated by cysteine scanning mutagenesis and measurements of accessibility through exposure to cysteine reagents. In the native protein, accessible residues were found only at the most extracellular residues of the 5th and 6th transmembrane segments (TMS) and the short loop between them. However, after modification by palytoxin a number of residues became accessible along the whole length of the 5th TMS and in the outer half of the 6th TMS, showing the contribution of each of these segments to the "channel" formed by the palytoxin-transformed Na,K-pump. Assuming that this structure is similar in the native and the palytoxin-transformed pump, our data allow us to determine the residues lining the cation pathway from the extracellular solution to their binding sites. A critical position in the 5th TMS contains a lysine conserved in all known nonelectrogenic H,K-ATPases, and a serine in all known electrogenic Na,K-ATPase sequences. Wild-type or mutant Na,K or H,K-ATPase a subunits were coinjected with the Bufo beta2 subunit in Xenopus oocytes and Rb(86) uptake and electrophysiological measurements were performed. An electrogenic activity was recorded for the H,K-ATPase mutants in which the positively charged lysine had been replaced by neutral or negatively charged residues, while nonelectrogenic transport was observed with the S(782)R mutant of the Na,K-ATPase. The presence or the absence of a positively charged residue at the S(782) position appears to be critical for the stoichiometry of cation exchange. PMID- 12763786 TI - Toward an understanding of ion transport through the Na,K-ATPase. AB - In the Na,K-ATPase the charge-translocating reaction steps were found to be binding of the third Na(+) ion to the cytoplasmic side and the release of all three Na(+) ions to the extracellular side as well as binding of the two K(+) ions on the extracellular side. The conformation transition E(1) --> E(2) was only of minor electrogenicity; all other reaction steps produced no significant charge movements. In the SR Ca-ATPase and the gastric H,K-ATPase, all ion-binding and -release steps were identified to move charge through the membrane. The high resolution structure of the SR Ca-ATPase in state E(1) revealed the position of the ion-binding sites in the transmembrane part of the protein. If the same arrangement is assumed for the Na pump, the missing expected charge movements in state E(1) may to be assumed to be apparent effects. With the proposal that binding of 2 Na(+) or 2 K(+) is compensated correspondingly by H(+) ions, agreement between structural and functional aspects is obtained. Investigations of the pH-dependence of ion-binding steps indicate competition between the ions and electrogenic H(+) binding in support of this concept. PMID- 12763787 TI - Na,K-pump reaction kinetics at the tip of a patch electrode: derivation of reaction kinetics for electrogenic and electrically silent reactions during ion transport by the Na,K-ATPase. AB - Patch-clamp electrophysiological techniques allow manipulations of electrochemical driving forces for ion transport by the Na,K-ATPase. For this reason, this technique has been used to study steady-state ion transport properties of the Na,K-ATPase. High temporal resolution during these manipulations also permits rapid reactions, such as extracellular ion-binding reactions, to be measured as charge movements when the enzyme is engaged in electroneutral ion exchange modes. Just as useful, but less widely recognized, is the ease with which electrophysiological techniques can be used to critically study reaction steps that do not directly involve ion binding. Three studies are briefly presented to show how pre-steady-state and/or steady-state electrophysiological techniques can be used to study ion-binding reactions in a novel fashion and the kinetics of electrically silent reaction steps of this enzyme. The reaction kinetics derived from each of these studies can be used to attain detailed mechanistic information about ion transport by the Na,K-ATPase. PMID- 12763789 TI - Binding of 1 Rb+ accelerates dephosphorylation of the Na+,K+-ATPase without leading to Rb+ occlusion. AB - In steady-state conditions and for concentrations of the K(+)-congener Rb(+) less than 2.5 mM, Rb(+)-dependent ATPase activity is significantly higher than the steady-state rate of breakdown of Rb(+)-occluded states, a discrepancy that disappears at sufficiently high [Rb(+)]. Direct experimental evidence is provided that supports the explanation that the binding of a single Rb(+) to the phosphoenzyme conformer E(2)P accelerates dephosphorylation without leading to the occlusion of the cation. PMID- 12763788 TI - Two-electrode voltage-clamp analysis of Na,K-ATPase asparagine 776 mutants. AB - Steady-state and pre-steady-state currents of Asn(776) mutants of Na,K-ATPase are presented. The stationary current generated by N776Q strongly depends on the membrane potential, but has a negative slope, opposite to that of the wild-type enzyme. The apparent rate constant of the reaction sequence E(1)P(Na(+)) <--> E(2)P + Na(+) of this mutant is rather independent of the membrane potential and is at resting and depolarizing membrane potential higher than that of the wild type enzyme. Thus, the voltage-dependent increase of the rate coefficient of the wild type that is associated with extracellular Na(+) rebinding is almost absent in the N776Q mutant. These findings indicate that dislocating the carboxamide group of Asn(776) decreases the affinity of sodium at its extracellular binding site. PMID- 12763790 TI - Kinetic investigations of the mechanism of the rate-determining step of the Na+,K+-ATPase pump cycle. AB - The kinetics of the E(2) --> E(1) conformational change of unphosphorylated Na(+),K(+)-ATPase from rabbit kidney were investigated via the stopped-flow technique using the fluorescent label RH421 (pH 7.4, 24 degrees C). The enzyme was preequilibrated in a solution containing 25 mM histidine and 0.1 mM EDTA to initially stabilize the E(2) conformation. On mixing enzyme with NaCl alone, tris ATP alone, or NaCl and tris-ATP simultaneously, a fluorescence decrease was observed. The reciprocal relaxation time, 1/tau, of the fluorescent transient was found to increase with increasing NaCl concentration and reached a saturating value in the presence of 1 mM tris-ATP of 54 (+/-3) s(-1). The experimental behavior could be described by a binding of Na(+) to the enzyme in the E(2) state with a dissociation constant of 31 (+/-7) mM, which induces a subsequent rate limiting conformational change to the E(1) state. Similar behavior, but with a decreased saturating value of 1/tau, was found when NaCl was replaced by choline chloride. Experiments performed with enzyme from shark rectal gland showed similar effects, but with a lower amplitude of the fluorescence change and a higher saturating value of 1/tau for both the NaCl and choline chloride titrations. The results suggest that Na(+) ions or salt in general play a regulatory role, similar to ATP, in enhancing the rate of the rate-limiting E(2) -> E(1) conformational transition by interaction with the E(2) state. PMID- 12763791 TI - Homology modeling of Na,K-ATPase: a putative third sodium binding site suggests a relay mechanism compatible with the electrogenic profile of Na+ translocation. AB - Identification of the third Na(+) binding site would be crucial in interpretation of the electrophysiological behavior of Na,K-ATPase. To address this question a three-dimensional homology model of Na,K-ATPase was built from the known crystallographic structure of Ca-ATPase (1EUL). Phe760, which is conserved in virtually all Ca-ATPases, is replaced by Ser768 in Na,K-ATPase, resulting in a small cavity between M4, M5, and M6. A partially hydrated Na(+) ion can be bound at this third site on the cytoplasmic side of cation binding sites 1 and 2. This leads to the proposal that the conductance of the "third Na(+)" ion across approximately 70% of the membrane dielectric may be achieved by adding up the passage of one Na(+) ion from the described cytoplasmic cavity to cation site 1 and the further conductance of the previously bound Na(+) ion from cation site 1 to the extracellular phase. This relay mechanism may therefore be compatible with the electrogenic profile of Na(+) translocation. PMID- 12763792 TI - Use of a fluorescent maleimide to probe structure-function relationships in stalk segments 4 and 5 of the yeast plasma-membrane H+-ATPase. AB - In the yeast plasma-membrane H(+)-ATPase and other P-type ATPases, conformational changes are transmitted between cytoplasmic and membrane-embedded domains via a stalk region composed of cytoplasmic extensions of membrane segments 2, 3, 4, and 5. The present study has used a fluorescent maleimide (Alexa-488) to probe Cys residues introduced into stalk segments 4 and 5 of the yeast enzyme. In the case of S5, Cys substitutions along one face led to a constitutive, 5- to 10-fold activation of the ATPase in the absence of glucose. Based on homology with SERCA Ca(2+)-ATPase, this face is likely to be buried in the interior of the protein, close to the P domain. Three Cys residues on the opposite face of S5 (A668C, S672C, and D676C) were accessible to Alexa-488 under all conditions tested. In addition, three other Cys residues at or near the boundary between the two faces reacted with Alexa-488 only (V665C, L678C) or preferentially (Y689C) in plasma membranes from glucose-metabolizing cells; this result provides the first direct evidence for a change in conformation of S5 during glucose activation. For stalk segment 4, site-directed mutagenesis gave no sign of a role in glucose-dependent regulation. Rather, substitutions at 13 consecutive positions along S4 caused kinetic changes consistent with a shift in equilibrium from E2 to E1. Four Cys residues along this stretch of S4 (Q357C, K362C, S364C, and S368C) reacted with Alexa-488, indicating that they are exposed to the aqueous medium as predicted in the SERCA-based structural model. PMID- 12763793 TI - The E1/E2-preference of gastric H,K-ATPase mutants. AB - Gastric H,K-ATPase has, in the absence of ATP and added ions, a preference for the E(2) conformation. Mutations in the cation-binding pocket often result in a preference for the E(1)-conformation. This can be paralleled by the occurrence of K(+)-independent ATPase activity. These two phenomena could be separated by combined mutagenesis of several residues in and around the cation-binding pocket. Models of the three-dimensional structure of H,K-ATPase visualize the relationship between the E(1)/E(2) preference and the structure. PMID- 12763794 TI - Nongastric H,K-ATPase: structure and functional properties. AB - Nongastric H,K-ATPases whose catalytic subunits (AL1) encoded by human ATP1AL1 and homologous animal genes comprise the third distinct group within the X,K ATPase family. No unique nongastric beta has been identified. Precise in situ colocalization and strong association of AL1 with beta1 of Na,K-ATPase was detected in apical membranes of rodent prostate epithelium. In this tissue, beta1NK serves as an authentic subunit of both the Na,K- and nongastric H,K pumps. Upon expression in Xenopus oocytes the human AL1 can assemble with beta1NK, and more efficiently with gastric betaHK, into functional H,K-pumps. Both AL1/beta complexes exhibit a similar K-affinity, and their K-transport depends on intra- and extracellular Na. These data provide new evidence that nongastric H,K-ATPase can perform Na/K-exchange, and indicate that beta does not significantly affect this ion-pump function. Analysis of human nongastric H,K ATPase expressed in Sf-21 insect cells revealed that AL1/betaHK exhibits substantial enzymatic activities in K-free medium and K stimulates, but Na has inhibitory effect on ATP hydrolysis. Thus, although the nongastric H,K-ATPase can function as Na/K exchanger, its reaction mechanism is different from that of the Na,K-ATPase. Human nongastric H,K-ATPase is highly sensitive to bufalin, digoxin, and digitoxin, but almost resistant to digoxigenin and ouabagenin. PMID- 12763795 TI - Mechanism of proton pumping by plant plasma membrane H+-ATPase: role of residues in transmembrane segments 5 and 6. AB - The mechanism of proton pumping by P-type plasma membrane H(+)-ATPases is not well clarified. Site-directed mutagenesis studies suggest that Asp684, situated in transmembrane segment M6, is involved in coordination of proton(s) in plant plasma membrane H(+)-ATPase. This hypothesis is supported by atomic models of H(+)-ATPases built on the basis of the crystal structure of the related SERCA1a Ca(2+)-ATPase. However, more biochemical, genetic, and structural studies are required before we will be able to understand the nature of the proton binding site(s) in P-type H(+)-ATPases and the mechanism of action of these pumps. PMID- 12763796 TI - Function and regulation of the two major plant plasma membrane H+-ATPases. AB - Plant plasma membrane H(+)-ATPases are encoded by a family of about ten genes organized into five subfamilies. Subfamilies I and II contain the most widely and highly expressed genes. In Nicotiana plumbaginifolia, they are represented, respectively, by pma2 (plasma membrane H(+)-ATPase) and pma4. When expressed in the yeast Saccharomyces cerevisiae, the two isoforms show different kinetics and are differently regulated by phosphorylation of the penultimate threonine residue and binding of regulatory 14-3-3 proteins. To determine if these differences also occurred in plant tissues, we developed an experimental approach allowing the characterization of a single isoform in the plant. When PMA2 bearing a 6-His tag was expressed under a strong transcription promoter in Nicotiana tabacum BY2 cells, solubilized from microsomal membranes and purified, the penultimate threonine was found to be phosphorylated, thus validating the model. PMID- 12763797 TI - Functional properties of the human copper-transporting ATPase ATP7B (the Wilson's disease protein) and regulation by metallochaperone Atox1. AB - Wilson's disease protein (WNDP) is a copper-transporting P(1)-type ATPase which plays a key role in normal distribution of copper in a number of tissues, particularly in the liver and the brain. Copper has numerous effects on WNDP, altering its structure, activity, and intracellular localization. To better understand the function of this copper-transporting ATPase and its regulation by copper, we have recently developed the functional expression systems for WNDP and for Atox1, a cytosolic protein that serves as an intracellular donor of copper for WNDP. Here we summarize the results of our experiments on characterization of the enzymatic properties of WNDP and the effects of Atox1 on the WNDP activity. PMID- 12763798 TI - Heavy metal transport CPx-ATPases from the thermophile Archaeoglobus fulgidus. AB - PIB-type ATPases transport diverse heavy metals (Cu(+), Ag(+), Cu(2+). Zn(2+), Cd(2+), Pb(2+), Co(2+)) across membranes. Toward understanding their mechanisms of metal selectivity, we are studying thermophilic archaeal PIB-type ATPases. Like other PIB ATPases, these are characterized by the presence of a cation binding CPX sequence in their 6th transmembrane segment and by cytoplasmic N terminus metal binding domains (N-MBDs). CopA and CopB from the thermophile Archaeoglobus fulgidus were cloned and expressed in E. coli. The resulting proteins were purified in a soluble active form. Typical yields were in the order of 3-5 mg of pure protein per liter of bacterial culture. Both enzymes showed maximum activity at 75-85 degrees C. CopA was activated by Ag(+)>Cu(+) while CopB was activated by Cu(2+)>Ag(+)>Cu(+). The differences in enzyme selectivity can be explained by different consensus sequences in the transmembrane cation binding domain (CopA: CPC, CopB: CPH). Mutagenesis studies show that the cysteines in the transmembrane CPC site of CopA are necessary for enzyme function, while those in the N-MBD (CXXC), although not essential, are required for maximum enzyme activity. Different from CopA, CopB has a His-rich N-MBD. Removal of this domain reduced enzyme activity without affecting enzyme selectivity. These studies show that these enzymes are an excellent system for structural functional studies directed to explain the mechanisms of metal selectivity by PIB ATPases. PMID- 12763799 TI - P-type ATPase superfamily: evidence for critical roles for kingdom evolution. AB - The P-type ATPase has become a protein superfamily. On the basis of sequence similarities, the phylogenetic analyses, and substrate specificities, this superfamily can be classified into 5 families and 11 subfamilies. A comparative phylogenetic analysis demonstrates the relationship between the molecular evolution of these subfamilies and the establishment of the kingdoms of living things. PMID- 12763800 TI - The Na,K-ATPase S5-H5 helix: structural link between phosphorylation and cation binding sites. PMID- 12763801 TI - Na,K-ATPase alpha-beta subunit interactions in the transmembrane domain. PMID- 12763802 TI - Negative changes of the membrane capacitance due to electrogenic Na transport by the Na,K-ATPase. PMID- 12763803 TI - Isolation of (alphabeta)4-tetraprotomer having half-of-sites ATP binding from solubilized dog kidney Na+/K+-ATPase. PMID- 12763804 TI - ATPase activity and oligomerization of solubilized Na+/K+-ATPase maintained by synthetic phosphatidylserine. PMID- 12763805 TI - Cation requirement for nucleotide binding to Na,K-ATPase. PMID- 12763806 TI - Single mutation of Lys or Arg residue in ATP binding pocket in rat Na/K-ATPase alpha-1 subunit induces different affinity change in high- and low-affinity ATP binding. PMID- 12763807 TI - Localization of catalytic active sites in the large cytoplasmic domain of Na+/K+ ATPase. PMID- 12763808 TI - Calorimetry of Na,K-ATPase. PMID- 12763809 TI - Expression of Na,K-ATPase in P. pastoris: Fe2+-catalyzed cleavage of the recombinant enzyme. PMID- 12763810 TI - The mechanism of Na-K interaction on Na,K-ATPase. PMID- 12763811 TI - Salt effects on the kinetics of the electrogenic Na+ transport in the Na,K ATPase. PMID- 12763812 TI - Mutational analysis of ouabain interaction with the M5-M6 hairpin of Na,K-ATPase. PMID- 12763813 TI - Role of the isoform-specific region of the Na,K-ATPase catalytic subunit. PMID- 12763814 TI - Structure/function analysis of Na,K-ATPase alpha1 and alpha2 central isoform specific regions reveals their involvement in regulation by protein kinase C. PMID- 12763815 TI - Characterization of the electrostatic component of the nucleotide binding to Na,K ATPase. PMID- 12763817 TI - Importance of Thr214 in the conserved TGES sequence of the Na+,K+-ATPase for vanadate binding and hydrolysis of E2P. PMID- 12763816 TI - Independent access of fluorescein isothiocyanate and Co(NH3)4ATP to their binding sites on the protomer of Na,K-ATPase. PMID- 12763819 TI - Mutational analysis of the interactions of the alpha and beta subunits of the Na,K-ATPase. PMID- 12763818 TI - Differential inactivation of Na,K-ATPase by erythrosin isothiocyanate. PMID- 12763820 TI - Mechanism of phosphoryl group transfer. PMID- 12763822 TI - K+ induced simultaneous liberation of two moles of Pi, one from one mole of EP and the other from EATP, of oligomeric H/K-ATPase from pig stomach. PMID- 12763821 TI - Oligomeric structure of P-type ATPases observed by single molecule detection technique. PMID- 12763823 TI - An improved method to measure the interactions of p-type ATPases with the lipidic environment. PMID- 12763824 TI - Extracellularly applied Br-TITU inhibits the Na+/K+ pump by interacting with tryptophan at the entrance to the cation sites. PMID- 12763825 TI - Interactions between cations and Na,K-ATPase membranes studied with solid-state NMR. PMID- 12763826 TI - Interaction between ATP and the Na/K-ATPase from duck supraorbital salt glands. PMID- 12763827 TI - Three-dimensional structure-activity relationship modeling of digoxin inhibition and docking to Na+,K+-ATPase. PMID- 12763829 TI - The sidedness of the direct route of occlusion of K+ in the Na+/K+-ATPase. PMID- 12763828 TI - A parallel study of eosin-fluorescence change and Rb+ occlusion in the Na+/K+ ATPase. PMID- 12763830 TI - The muscle-specific beta m protein is functionally different from other members of the X,K-ATPase beta-subunit family. PMID- 12763831 TI - Influence of intramembrane electric charge on H,K-ATPase. PMID- 12763832 TI - The role of Lys791 and Asn792 in gastric H,K-ATPase. PMID- 12763833 TI - Functional consequences of charge reversals of acidic residues in M1 of the SR Ca ATPase. PMID- 12763834 TI - Overexpression of SERCA1a Ca2+-ATPase in yeast. PMID- 12763835 TI - TNP-8N3-ADP photoinactivation of the phosphatase activity of FITC-modified SERCA. PMID- 12763836 TI - Ca2+ occlusion of sarcoplasmic reticulum Ca2+-ATPase by CrATP. PMID- 12763837 TI - A model accounting for the simultaneous transport of calcium and manganese in sarcoplasmic reticulum membranes. PMID- 12763839 TI - Time-resolved partial reactions of the SR Ca-ATPase investigated with a fluorescent styryl dye. PMID- 12763838 TI - Interaction of an aromatic dibromo-isothiouronium derivative with the Ca-ATPase of sarcoplasmic reticulum. PMID- 12763840 TI - Macrocyclic carbon suboxide oligomers as potent inhibitors of the Na,K-ATPase. PMID- 12763841 TI - The inherent energy in SR Ca2+-ATPase is convertible into chemical work. PMID- 12763842 TI - Purification of SERCA1a Ca2+-ATPase mutants expressed in yeast. PMID- 12763843 TI - Mutational analysis of Lys252 and its interaction with loop 6-7 in the SR Ca2+ ATPase. PMID- 12763844 TI - Phospholamban inhibits Ca2+ pump oligomerization and intersubunit free energy exchange leading to activation of cardiac muscle SERCA2a. PMID- 12763846 TI - The nature of the low-frequency normal modes of the E1Ca form of the SERCA1 Ca2+ ATPase. PMID- 12763847 TI - Protonation of the Neurospora crassa plasma membrane H+-ATPase as a function of pH monitored by ATR-FTIR. PMID- 12763845 TI - ATP regulation of calcium binding in Ca2+-ATPase molecules of the sarcoplasmic reticulum. PMID- 12763848 TI - Mutagenic study of residues in transmembrane helix 4, 5, and 6 of the plant plasma membrane P-type H+-ATPase. PMID- 12763849 TI - Mutational analysis of charged residues in the putative KdpB-TM5 domain of the Kdp-ATPase of Escherichia coli. PMID- 12763850 TI - Functional roles of the alpha isoforms of the Na,K-ATPase. AB - The Na,K-ATPase is composed of two subunits, alpha and beta, and each subunit consists of multiple isoforms. In the case of alpha, four isoforms, alpha1, alpha2, alpha3, and alpha4 are present in mammalian cells. The distribution of these isoforms is tissue- and developmental-specific, suggesting that they may play specific roles, either during development or coupled to specific physiological processes. In order to understand the functional properties of each of these isoforms, we are using gene targeting, where animals are produced lacking either one copy or both copies of the corresponding gene or have a modified gene. To date, we have produced animals lacking the alpha1 and alpha2 isoform genes. Animals lacking both copies of the alpha1 isoform gene are not viable, while animals lacking both copies of the alpha2 isoform gene make it to birth, but are either born dead or die very soon after. In the case of animals lacking one copy of the alpha1 or alpha2 isoform gene, the animals survive and appear healthy. Heart and EDL muscle from animals lacking one copy of the alpha2 isoform exhibit an increase in force of contraction, while there is reduced force of contraction in both muscles from animals lacking one copy of the alpha1 isoform gene. These studies indicate that the alpha1 and alpha2 isoforms carry out different physiological roles. The alpha2 isoform appears to be involved in regulating Ca(2+) transients involved in muscle contraction, while the alpha1 isoform probably plays a more generalized role. While we have not yet knocked out the alpha3 or alpha4 isoform genes, studies to date indicate that the alpha4 isoform is necessary to maintain sperm motility. It is thus possible that the alpha2, alpha3, and alpha4 isoforms are involved in specialized functions of various tissues, helping to explain their tissue- and developmental-specific regulation. PMID- 12763851 TI - Ion pump-interacting proteins: promising new partners. AB - The sorting and regulation of the Na,K and H,K-ATPases requires that the pump proteins must associate, at least transiently, with kinases, phosphatases, scaffolding molecules, and components of the cellular trafficking machinery. The identities of these interacting proteins and the nature of their associations with the pump polypeptides have yet to be elucidated. We have begun a series of yeast two-hybrid screens employing structurally defined segments of pump polypeptides as baits in order to gain insight into the nature and function of these interacting proteins. PMID- 12763852 TI - Amino acids in the TM4-TM5 loop of Na,K-ATPase are important for biosynthesis. AB - The ten-transmembrane Na,K-ATPase alpha-subunit exposes very few amino acids to the extra membrane space except for an approximately 408 residue-long loop between transmembrane segments four and five. The present paper focuses on the role of this loop in biosynthesis of functional Na,K-ATPase. Expression of 39 mutations in this loop to phylogenetically conserved as well as nonconserved residues showed that only two could be expressed at 30 degrees C. By contrast, only five could not be produced in a functional form at 15 degrees C. A detailed analysis showed that a number of these mutants are temperature-sensitive folding mutants, as they induce the unfolded protein response at 30 degrees C but not at 15 degrees C. We used an algorithm to predict that residues (868)ENGFLIPIHLL(878) in the L78 loop exposed to the endoplasmic reticulum lumen constitute the most likely BiP binding site. Correct folding of this sequence may be important in the endoplasmic reticulum quality control, as the same loop is responsible for the alpha-beta-associations required to leave this compartment. On the basis of the Ca-ATPase crystal structure and the presented data, we propose a model to account for the role of the TM4-TM5 loop in Na,K-ATPase biosynthesis. PMID- 12763853 TI - Differential degradation of the Na+/K+-ATPase subunits in the plasma membrane. AB - The cell-surface expression of the Na(+)/K(+)-ATPase is tightly regulated at the ER/Golgi transit level. The assembly of the alpha- and beta-subunits is a prerequisite for the maturation of the enzyme and its exit from the ER. At present, the fate of the functional ATPase, once it reaches the plasma membrane, is obscure, although endocytosis, recycling, and degradation in the lysosome are known to be involved. In this study, we shall demonstrate the differential degradation of the Na(+)/K(+)-ATPase subunits in the plasma membrane: that is, lysosomal degradation of alpha-subunits and proteasomal degradation of beta subunits, and propose the possibility of the subunit dissociation in the plasma membrane. PMID- 12763854 TI - FXYD proteins as regulators of the Na,K-ATPase in the kidney. AB - The FXYD gene family has seven members in mammals and others in fish. Five of these (FXYD1, FXYD2, FXYD4, FXYD7, and PLMS from shark) have been shown to alter the activity of the Na,K-ATPase, as described by other papers in this volume. The gene structure of FXYD family members suggests assembly from protein domain modules and gene duplication. The gamma subunit is unique in the family for having alternative splice variants in the coding region and can be posttranslationally modified with different final consequences for enzyme properties. The nonoverlapping distribution of gamma and CHIF (FXYD4) in kidney helps to explain physiological differences in Na(+) affinity among nephron segments. We also detected phospholemman (FXYD1) in kidney. By immunofluorescence, it was found in extraglomerular mesangial cells (EM cells) of the juxtaglomerular apparatus and in the afferent arteriole. Contrary to many reports that only alpha1 and beta1 are expressed in the kidney, we found that alpha2 and beta2 are present, although not in any nephron segment. Both were detected in arterioles, and beta2 was found in the EM cells. In contrast, alpha1, beta1, and gamma were found in adjacent macula densa. Phospholemman, alpha2, and beta2 are proposed to have distinct roles in regulating the sodium pump in structures involved in tubuloglomerular feedback. PMID- 12763855 TI - FXYD proteins: new tissue- and isoform-specific regulators of Na,K-ATPase. AB - The recently defined FXYD protein family contains seven members that are small, single-span membrane proteins characterized by a signature sequence containing an FXYD motif and three other conserved amino acid residues. Until recently, the functional role of FXYD proteins was largely unknown, with the exception of the gamma subunit of Na,K-ATPase, which was shown to be a specific regulator of renal alpha1-beta1 isozymes. We have investigated whether other members of the FXYD family may have a similar role as the gamma subunit and have found that CHIF (corticosteroid hormone-induced factor, FXYD4), FXYD7, as well as phospholemman (FXYD1) specifically associate with Na,K-ATPase and preferentially with alpha1 beta isozymes in native tissues, and produce distinct effects on the transport properties of Na,K-ATPase that are adapted to the physiological demands of the tissues in which they are expressed. These results provide evidence for a unique and novel mode of regulation of Na,K-ATPase by FXYD proteins that involves a tissue-specific expression of an auxiliary subunit of distinct Na,K-ATPase isozymes. PMID- 12763856 TI - A specific functional interaction between CHIF and Na,K-ATPase: role of FXYD proteins in the cellular regulation of the pump. AB - CHIF (corticosteroid hormone-induced factor) is a member of the FXYD family that shares approximately 50% homology with the gamma subunit of Na,K-ATPase. It is expressed in renal collecting duct and distal colon, and is upregulated by Na(+) deprivation and high K(+) diet. Both CHIF and gamma are coimmunoprecipitated by an anti-alpha subunit antibody, and alpha is immunoprecipitated by anti-gamma and anti-CHIF antibodies. (86)Rb(+) flux experiments in CHIF-transfected HeLa cells demonstrate that CHIF increases the affinity for cytoplasmic Na(+), but does not affect the affinity for extracellular K(Rb). A physiological role of CHIF in kidney function is further elucidated by the phenotypic analysis of CHIF knockout mice. Taken together with data by others, it appears that FXYD proteins are tissue-specific subunits or regulators of the Na,K-ATPase whose function is to adjust the pump kinetics to particular physiological needs. PMID- 12763857 TI - Immunocytochemical localization of Na,K-ATPase gamma subunit and CHIF in inner medulla of rat kidney. AB - The gamma subunit of Na,K-ATPase and CHIF both belong to the FXYD single-membrane spanning protein family and have been suggested to have regulatory functions in kidney tubules. CHIF is known to be present in the collecting duct, and gamma has been demonstrated in several segments of the rat kidney tubule, but never clearly in the inner medullary collecting duct (IMCD). Here, we demonstrate the cellular and subcellular localization of the gamma subunit and CHIF in the IMCD in inner medulla by using Western blotting, laser-scanning confocal immunofluorescence, and immunoelectron microscopy. In the initial quarter of the IMCD (next to the outer medulla), antibodies against the C-terminal of gamma as well as splice variant gammaa labeled the basolateral surface of intercalated cells (ICs), while principal cells (PCs) remained unlabeled. In the middle segment of the IMCD, all PCs exhibited distinct basolateral staining for the gammaC-terminal as well as gammaa and CHIF. Immunoelectron microscopy showed that the gammaC-terminal and CHIF were associated with the inner leaflet of the basolateral plasma membrane in the labeled cells. Immunoblotting demonstrated the presence of both the gammaC terminal and gammaa in inner medullary tissue. However, splice variant gammab was not detected in inner medulla by immunocytochemistry or immunoblotting. The present observations demonstrate that the Na,K-ATPase gamma subunit and CHIF are strategically located in the inner medulla to participate in the fine-tuning of urine ion composition through the regulation of the Na,K-ATPase activity in the IMCD. PMID- 12763858 TI - Adaptation of murine inner medullary collecting duct (IMCD3) cell cultures to hypertonicity. AB - Recently, we have adapted IMCD3 cell cultures to survive under increasing hypertonic conditions (i.e., 600 and 900 mOsmol/kg H(2)O). In adapted cells, ATPase activity is increased by one order of magnitude, while the expression of the alpha and beta subunit is increased by a factor of 4 to 5 over controls (300 mOsmol/kg H(2)O). Corresponding increases in mRNAs were also detected. The gamma subunit has been described as being uniquely expressed in some areas of the kidney, but never in cell cultures (even those derived from kidney tissues). However, the gamma subunit was detected at the protein and mRNA levels in the adapted IMCD3 cells. In contrast to the alpha and beta subunits, the levels of gamma protein and mRNA expression continue to increase as a function of the media ion concentration. We have also demonstrated that signaling pathways that upregulate the alpha, beta, and gamma subunits are very different. Increasing concentrations of the PI3 kinase inhibitor, LY294002, resulted in a dose dependent reduction in the expression of the gamma subunit, with total abolition at 10 micro M. However, LY294002 had no significant effect on the expression of the alpha subunit. Inhibition of the JNK2 (but not of the JNK1) pathways by dominant negative transfections abolished the upregulation of the gamma, but not the a subunit. Failure to upregulate the expression of the gamma subunit was associated with a marked decrease in cell viability upon stress. PMID- 12763859 TI - Gamma structural variants differentially regulate Na,K-ATPase properties. AB - Renal Na,K-ATPase is tightly bound to a small regulatory protein, the gamma subunit (FXYD2). In rat, it occurs in two splice variants with different N termini. Immunolocalization on kidney sections revealed distinct distribution of the gamma splice variants along the rat nephron. Where coexpressed, they coimmunoprecipitated with each other along with the alpha subunit, suggesting assembly in oligomeric complexes. Functional consequences of association with gamma were monitored in stably transfected NRK-52E cells. The outcome was that splice variants can differentially modulate the major intrinsic properties of the Na,K-ATPase under normal and stress-related conditions. The data imply an adaptive physiological mechanism of regulation of renal Na,K-ATPase through modulation of pump properties, gene expression, or both. PMID- 12763860 TI - Structure/function studies of the gamma subunit of the Na,K-ATPase. AB - The Na,K-ATPase gamma subunit is present primarily in kidney as two splice variants, gammaa and gammab, which differ only at their extracellular N-termini. Two distinct effects of gamma are seen in biochemical Na,K-ATPase assays of mammalian (HeLa) cells transfected with gammaa or gammab, namely, (i) a decrease in K'(ATP) probably secondary to a shift in steady-state E(1) <--> E(2) poise in favor of E(1) and (ii) an increase in cytoplasmic K(+)/Na(+) antagonism seen as an increase in K'(Na) at high K(+) concentration. Mutagenesis experiments involving alterations in extramembranous regions of gamma indicate that different regions mediate the aforementioned distinct effects and that the effects appear to be long range. Studies of ouabain-sensitive fluxes with intact cells confirm the gamma effects seen with membranes and also suggest an additional effect (increase) in apparent affinity for extracellular K(+). Alteration in gamma function was also evidenced in the behavior of a G41 -->R mutation within the transmembrane domain of gamma. G41R is associated with autosomal dominant renal magnesium wasting. Our studies show that this mutation in the gammab variant retards trafficking of gamma, but not alphabeta pumps, to the cell surface and abolishes functional effects of gamma, consistent with the conclusion that the Mg(2+) transport defect is secondary to loss of gamma modulation of Na,K-ATPase function. PMID- 12763861 TI - Cell-specific expression of three members of the FXYD family along the renal tubule. AB - The gamma subunit of Na/K/ATPase is a small membrane protein that shares homologies with other members of the FXYD family, like phospholemman and CHIF (corticosteroid hormone-induced factor). Both the gamma subunit and CHIF modulate sodium pump properties. The gamma subunit increases the apparent affinity of the pump for ATP and reduces its apparent affinity for sodium. CHIF, in contrast, augments its apparent affinity for sodium. Gamma subunit expression is essentially restricted to the kidney, with two main splice variants, gammaa and gammab, which differ only at their extracellular N-termini. We have investigated in detail the cell-specific expression of the two splice variants of gamma within the kidney and compared it to that of CHIF. While both gamma variants affect catalytic properties of the pump (without detectable difference between a and b forms), their localization along the nephron is partially distinct. Both variants are coexpressed in the proximal tubule and in the medullary part of the thick ascending limb of Henle's loop (TAL). In contrast, their expression differs in the downstream tubular segments. Within the renal cortex, the sole gamma a variant was found in macula densa cells and in principal cells of the initial parts of the collecting duct. Gamma b is in the cortical part of the TAL. Outer and inner medullary collecting ducts lack detectable gamma expression. These latter nephron segments express CHIF, and no overlap between gamma and CHIF expression along the nephron was observed. Such distinct cell-specific expression argues for complementary roles to modulate Na/K/ATPase activity. PMID- 12763862 TI - Dominant isolated renal magnesium loss is caused by misrouting of the Na+,K+ ATPase gamma-subunit. AB - Hereditary primary hypomagnesemia comprises a clinically and genetically heterogeneous group of disorders in which hypomagnesemia is due to either renal or intestinal Mg(2+) wasting. These disorders share the general symptoms of hypomagnesemia, tetany and epileptiformic convulsions, and often include secondary or associated disturbances in calcium excretion. In a large Dutch family with autosomal dominant renal hypomagnesemia, associated with hypocalciuria, we mapped the disease locus to a 5.6-cM region on chromosome 11q23. After candidate screening, we identified a heterozygous mutation in the FXYD2 gene, encoding the Na(+),K(+)-ATPase gamma-subunit, cosegregating with the patients of this family, which was not found in 132 control chromosomes. The mutation leads to a G41R substitution, introducing a charged amino acid residue in the predicted transmembrane region of the gamma-subunit protein. Expression studies in insect Sf9 and COS-1 cells showed that the mutant gamma-subunit protein was incorrectly routed and accumulated in perinuclear structures. In addition to disturbed routing of the G41R mutant, Western blot analysis of Xenopus oocytes expressing wild-type or mutant gamma-subunit showed mutant gamma subunit lacking a posttranslational modification. Finally, we investigated two individuals lacking one copy of the FXYD2 gene and found their serum Mg(2+) levels to be within the normal range. We conclude that the arrest of mutant gamma subunit in distinct intracellular structures is associated with aberrant posttranslational processing and that the G41R mutation causes dominant renal hypomagnesemia associated with hypocalciuria through a dominant negative mechanism. PMID- 12763863 TI - FXYD7, the first brain- and isoform-specific regulator of Na,K-ATPase: biosynthesis and function of its posttranslational modifications. AB - The FXYD protein family has recently been defined as a result of the search for homologues of the Na,K-ATPase gamma subunit, CHIF, and phospholemman in EST and gene data banks. FXYD7 has been seen to have a role as a brain- and isozyme specific regulator of Na/K-ATPase. In this study, the biosynthesis, membrane topology, nature, and role of the processing of FXYD7 are investigated. PMID- 12763864 TI - Phosphorylation of the Na+,K+-ATPase in skeletal muscle: potential mechanism for changes in pump cell-surface abundance and activity. AB - In skeletal muscle, insulin stimulation leads to phosphorylation of Na(+),K(+) ATPase alpha-subunits on both serine/threonine and tyrosine residues, translocation of Na(+),K(+)-ATPase molecules to the plasma membrane, and increased Na(+),K(+)-ATPase activity. The molecular nature of the tyrosine kinase that phosphorylates Na(+),K(+)-ATPase is not yet identified. In vitro phosphorylation experiments show that the alpha-subunit of Na(+),K(+)-ATPase from skeletal muscle is a substrate for the tyrosine-specific protein kinase c-src. Tyrosine phosphorylation of the alpha-subunits of Na(+),K(+)-ATPase may be an important mechanism for insulin-mediated regulation of Na(+),K(+)-ATPase translocation and activity. PMID- 12763865 TI - Physiological functions of plasma membrane and intracellular Ca2+ pumps revealed by analysis of null mutants. AB - It is known that plasma membrane Ca(2+)-transporting ATPases (PMCAs) extrude Ca(2+) from the cell and that sarco(endo)plasmic reticulum Ca(2+)-ATPases (SERCAs) and secretory pathway Ca(2+)-ATPases (SPCAs) sequester Ca(2+) in intracellular organelles; however, the specific physiological functions of individual isoforms are less well understood. This information is beginning to emerge from studies of mice and humans carrying null mutations in the corresponding genes. Mice with targeted or spontaneous mutations in plasma membrane Ca(2+)-ATPase isoform 2 (PMCA2) are profoundly deaf and have a balance defect due to the loss of PMCA2 in sensory hair cells of the inner ear. In humans, mutations in SERCA1 (ATP2A1) cause Brody disease, an impairment of skeletal muscle relaxation; loss of one copy of the SERCA2 (ATP2A2) gene causes Darier disease, a skin disorder; and loss of one copy of the SPCA1 (ATP2C1) gene causes Hailey-Hailey disease, another skin disorder. In the mouse, SERCA2 null mutants do not survive to birth, and heterozygous SERCA2 mutants have impaired cardiac performance and a high incidence of squamous cell cancers. SERCA3 null mutants survive and appear healthy, but endothelium-dependent relaxation of vascular smooth muscle is impaired and Ca(2+) signaling is altered in pancreatic beta cells. The diversity of phenotypes indicates that the various Ca(2+) transporting ATPase isoforms serve very different physiological functions. PMID- 12763866 TI - Characterization of PISP, a novel single-PDZ protein that binds to all plasma membrane Ca2+-ATPase b-splice variants. AB - Plasma membrane Ca(2+) ATPases (PMCAs) maintain intracellular Ca(2+) homeostasis and participate in the local regulation of Ca(2+) signaling. Spatially separate demands for Ca(2+) regulation require proper membrane targeting of PMCAs, but the mechanism of PMCA targeting is unknown. Using the PMCA2b carboxyl-terminal tail as yeast two-hybrid bait, we isolated a novel PDZ domain-containing protein from a human brain cDNA library. This protein, named PISP for PMCA-interacting single PDZ protein, consists of 140 amino acids and contains little else besides a single PDZ domain. Pulldown experiments showed that PISP interacts with all PMCA b-splice forms. PISP was found to be ubiquitously expressed and, in MDCK cells, was present in a punctate pattern throughout the cytosol and at the basolateral membrane. When added to microsomal membranes expressing PMCA4b, PISP was unable to stimulate the PMCA-dependent ATPase activity. Our data suggest that PISP is a transiently interacting partner of the PMCA b-splice forms that may play a role in their sorting to or from the plasma membrane. PMID- 12763868 TI - The thermogenic function of the sarcoplasmic reticulum Ca2+-ATPase of normal and hyperthyroid rabbit. AB - After formation of a Ca(2+) gradient, the amount of heat released during the hydrolysis of each mol of ATP cleaved (DeltaH(cal)) varies depending on the Ca(2+)-ATPase isoform expressed by the muscle cell. In vesicles derived from the sarcoplasmic reticulum of white muscle (SERCA 1) most of the ATP cleaved is not coupled to Ca(2+) transport, and the DeltaH(cal) varies between -20 and -22 kcal/mol. In contrast, in vesicles derived from red muscle (SERCA 2a) the hydrolysis of ATP is coupled with Ca(2+) transport, and the DeltaH(cal) varies between -12 and -14 kcal/mol. Hyperthyroidism increases the rate of heat production by the Ca(2+)-ATPase fourfold in white muscle and 40-fold in red muscle. In hyperthyroid rabbits, the amount of sarcoplasmic reticulum protein recovered from white and red muscle is four- to fivefold greater than that obtained from control rabbits. Hyperthyroid red muscle expresses SERCA 1, and the vesicles derived from these muscle hydrolyze ATP through a catalytic route that is not coupled to Ca(2+) transport, thus increasing the amount of heat released during ATP hydrolysis, the DeltaH(cal) varying between -20 and -22 kcal/mol. PMID- 12763867 TI - The regulation of SERCA-type pumps by phospholamban and sarcolipin. AB - Both sarcolipin (SLN) and phospholamban (PLN) lower the apparent affinity of either SERCA1a or SERCA2a for Ca(2+). Since SLN and PLN are coexpressed in the heart, interactions among these three proteins were investigated. When SERCA1a or SERCA2a were coexpressed in HEK-293 cells with both SLN and PLN, superinhibition resulted. The ability of SLN to elevate the content of PLN monomers accounts, at least in part, for the superinhibitory effects of SLN in the presence of PLN. To evaluate the role of SLN in skeletal muscle, SLN cDNA was injected directly into rat soleus muscle and force characteristics were analyzed. Overexpression of SLN resulted in significant reductions in both twitch and tetanic peak force amplitude and maximal rates of contraction and relaxation and increased fatigability with repeated electrical stimulation. Ca(2+) uptake in muscle homogenates was impaired, suggesting that overexpression of SLN may reduce the sarcoplasmic reticulum Ca(2+) store. SLN and PLN appear to bind to the same regulatory site in SERCA. However, in a ternary complex, PLN occupies the regulatory site and SLN binds to the exposed side of PLN and to SERCA. PMID- 12763869 TI - Na,K-ATPase as a signal transducer. AB - Recent studies have indicated that Na,K-ATPase may, in addition to being the key regulator of intracellular Na(+) and K(+) concentration, act as a signal transducer. Despite extensive research, the biological role for ouabain, a natural ligand of Na,K-ATPase, is not well understood. We have reported that exposure of rat proximal tubular cells (RPTC) to doses of ouabain that inhibit the Na,K-ATPase activity by less than 50% (10 nM - 500 micro M), will induce intracellular [Ca(2+)](i) oscillations and that this calcium signal leads to activation of the transcription factor NF-kappaB. The ouabain-induced calcium oscillations were blocked by an inhibitor of the IP(3) receptors but not by phospholipase C inhibitors nor by cellular depletion of IP(3), suggesting that the calcium signal is not due to phospholipase C-mediated IP(3) release. Fluorescence resonance energy transfer (FRET) studies suggested a close proximity between the Na,K-ATPase and IP(3) receptor. Our findings demonstrate a novel principle for calcium signaling via Na,K-ATPase. PMID- 12763870 TI - Molecular mechanisms of Na/K-ATPase-mediated signal transduction. AB - Our recent work shows that in addition to pumping ions, Na/K-ATPase acts as a signal transducer. Binding of ouabain to Na/K-ATPase changes the interaction of the enzyme with neighboring membrane proteins and induces the formation of multiple signaling modules, resulting in activation of Src, transactivation of the EGF receptor (EGFR), and increased production of reactive oxygen species (ROS). Interaction of these signals leads to activation of several other cascades, including p42/44 and p38 MAPKs, phospholipase C, and protein kinase C isozymes, in a cell-specific manner. Ouabain also increases [Ca(2+)](i) and contractility, induces some of the early-response protooncogenes, and activates transcription factors AP-1 and NF-kappaB. Interplay among these pathways eventually results in changes in the expression of a number of growth-related genes and in cell growth. Significantly, inhibition of Src blocked many of the aforementioned ouabain-activated signaling pathways. Furthermore, Src binds to Na/K-ATPase directly and ouabain regulates the interaction between Src and the enzyme, resulting in Src activation. To address the possibility that the signaling Na/K-ATPase is concentrated in a separate pool on the plasma membrane, we have assessed interaction of the enzyme with caveolins. These studies indicated that Na/K-ATPase was concentrated in caveolae/rafts. In addition, caveolin-1 can be co-immunoprecipitated with Na/K-ATPase. Finally, we have shown that the signaling function of the enzyme is also pivotal to ouabain-induced nongenomic effects on cardiac myocytes. PMID- 12763871 TI - Low concentrations of ouabain activate vascular smooth muscle cell proliferation. AB - In vascular smooth muscle cells the sodium pump complex can act as an intracellular signal transducing complex activated by low ouabain concentrations, which inhibit sufficient pumps to activate a transduction cascade via transactivation of EGFR, but insufficient pumps to alter intracellular ions. Higher concentrations interfere with proliferation. This biphasic ouabain response occurs in human, canine, and rat VSMC at concentrations that reflect the differing ouabain affinities of the alpha1 isoforms of the three species. This supports the proposal that this effect occurs via ouabain binding to the alpha1 subunit of the Na pump. These data suggest a new transducing function of ouabain Na pump interaction, distinct from the cellular ionic effects resulting from pump inhibition. This transducing function occurs at ouabain concentrations that do not perturb cytoplasmic ion content and requires specific localization of pumps to caveolae. PMID- 12763872 TI - Regulation of Ca2+ signaling by Na+ pump alpha-2 subunit expression. AB - The role of the Na(+) pump alpha2 subunit in Ca(2+) signaling was examined in primary cultured astrocytes from wild type (WT) mouse fetuses and those with a null mutation in one (Het) or both (KO) alpha2 genes. Cytosol Na(+) and Ca(2+) concentrations ([Na(+)](CYT) and [Ca(2+)](CYT)) were measured with benzofuran isophthalate (SBFI) and Fura-2. Het astrocytes express approximately 50% of normal alpha2; KO cells express none. Resting [Na(+)](CYT) = 6.5 mM in WT, 6.8 mM in Het, and 8.0 mM in KO cells; 500 nM ouabain (which inhibits only alpha2) equalized [Na(+)](CYT) at 8 mM in all genotypes. Resting [Ca(2+)](CYT) = 132 nM in WT, 162 nM in Het and 196 nM in KO cells. Cyclopiazonic acid (CPA), which inhibits endoplasmic reticulum (ER) Ca(2+) pumps, induces elevation of [Ca(2+)]CYT. These Ca(2+) responses are augmented in Het and KO cells. This correlates with alpha2 Na(+) pump and Na(+)/Ca(2+) exchanger (NCX) localization in plasma membrane (PM) microdomains that overlie the ER. Selective reduction of alpha2 Na(+) pump activity apparently elevates local [Na(+)] and, via NCX, [Ca(2+)] in the tiny cytosolic space between the PM and ER. This augments adjacent ER Ca(2+) stores and amplifies Ca(2+) signaling without elevating bulk [Na(+)](CYT). PMID- 12763873 TI - Visualization of Na,K-ATPase interacting proteins using FRET technique. AB - Signaling transduction mediated by protein aggregates within specific microdomains has been receiving increased attention. We previously showed that Na,K-ATPase, partially inhibited by ouabain, induces intracellular calcium (Ca(2+)) oscillations which involve Ca(2+) release from the endoplasmic reticulum (ER). Plasma membrane bound Na,K-ATPase and proteins in the ER are in close proximity to each other, and signal transduction may occur via a physical interaction or a microdomain. To study these signaling pathways and intricate microenvironments, sophisticated methods are required. One way to detect molecular interactions in the nanometer scale (1-10 nm) is fluorescence resonance energy transfer (FRET). Thus, FRET provides vital insight into the action of Na,K ATPase to trigger intracellular signaling events. PMID- 12763874 TI - Na/K-ATPase regulates intracellular ROS level in cerebellum neurons. PMID- 12763876 TI - Molecular activity of Na+,K+-ATPase relates to the packing of membrane lipids. PMID- 12763875 TI - Intermolecular Interaction between Na+/K+-ATPase alpha subunit and glycogen phosphorylase. PMID- 12763877 TI - Proteins binding to alpha1beta1 isozyme of Na,K-ATPase. PMID- 12763879 TI - Defining the nature and sites of interaction between FXYD proteins and Na,K ATPase. PMID- 12763878 TI - Domains involved in the interactions between FXYD and Na,K-ATPase. PMID- 12763880 TI - NO regulation of Na,K-ATPase: nitric oxide regulation of the Na,K-ATPase in physiological and pathological states. PMID- 12763881 TI - Functional expression of the alpha4 isoform of the Na,K-ATPase in both diploid and haploid germ cells of male rats. PMID- 12763882 TI - Responses at the translational level to heterologous expression of the Na,K ATPase. PMID- 12763883 TI - Protein kinase C phosphorylation directed at novel C-terminal sites in Na,K ATPase. PMID- 12763884 TI - Modification of the PKC phosphorylation site Ser-23 of the rat alpha1 subunit. PMID- 12763885 TI - Expression of a Na,K-ATPase-EGFP chimera in COS cells: can internalization explain PKA- or PKC-mediated inhibition of 86Rb uptake? PMID- 12763886 TI - PKA and PKC phosphorylation of gastric H,K-ATPase. PMID- 12763887 TI - Seasonal changes of Ca-ATPase activity in skeletal muscle sarcoplasmic reticulum of the ground squirrel Spermophilus undulatus. PMID- 12763888 TI - Acidic-lipid responsive regions of the plasma membrane Ca2+ pump. PMID- 12763889 TI - Short-term aldosterone action on Na,K-ATPase surface expression: role of aldosterone-induced SGK1? AB - Aldosterone controls extracellular volume and blood pressure by regulating Na(+) reabsorption across epithelial cells of the aldosterone-sensitive distal nephron (ASDN). This effect is mediated by a coordinate action on the luminal channel ENaC (generally rate limiting) and the basolateral Na,K-ATPase. Long-term effects of aldosterone (starting within 3 to 6 hours and increasing over days) are mediated by the direct and indirect induction of stable elements of the Na(+) transport machinery (e.g., Na,K-ATPase alpha subunit), whereas short-term effects appear to be mediated by the upregulation of short-lived elements of the machinery (e.g., ENaC alpha subunit) and of regulatory proteins, such as the serum- and glucocorticoid-regulated kinase SGK1. We have recently shown that in cortical collecting duct (CCD) from adrenalectomized (ADX) rats, the increase in Na,K-ATPase activity (approximately threefold in 3 h), induced by a single aldosterone injection, can be fully accounted for by the increase in Na,K-ATPase cell-surface expression. Using the model cell line mpkCCD(cl4), we showed that the parallel increase in Na,K-ATPase function [assessed by Na(+) pump current (I(p)) measurements] and cell-surface expression depends on transcription and translation, and that it is not secondary to a change in apical Na(+) influx. As a first approach to address the question whether the aldosterone-induced regulatory protein SGK1 might play a role in mediating Na,K-ATPase translocation, we have used the Xenopus laevis expression system. SGK1 coexpression indeed increased both the Na(+) pump current and the surface expression of pumps containing the rat alpha1 subunits. In summary, aldosterone controls Na(+) reabsorption in the short term not only by regulating the apical cell-surface expression of ENaC but also by coordinately acting on the basolateral cell surface expression of the Na,K-ATPase. Results obtained in the Xenopus oocyte expression system suggest the possibility that this effect could be mediated in part by the aldosterone-induced kinase SGK1. PMID- 12763890 TI - Renal tubule sodium transporter abundance profiling in rat kidney: response to aldosterone and variations in NaCl intake. AB - Based on extensive physiological study of sodium transport mechanisms along the renal tubule, complementary DNAs for all of the major transporters and channels responsible for renal tubular sodium reabsorption have been cloned over the past decade. There is now a comprehensive set of cDNA and antibody probes that can be used to investigate physiological mechanisms on a molecular level. Using rabbit polyclonal antibodies to all of the major renal Na transport proteins, we have developed profiling methods allowing comprehensive, integrated analysis of sodium transporter protein abundance changes along the renal tubule in response to physiological and pathophysiological perturbations. Here, we review some of our recent findings with this approach, focusing on renal responses to aldosterone and to variations in NaCl intake. PMID- 12763891 TI - Mechanism of control of Na,K-ATPase in principal cells of the mammalian collecting duct. AB - The collecting duct is the site of final Na reabsorption according to Na balance requirements. Using isolated rat cortical collecting ducts (CCD) and mpkCCD(cl4) cells, a mouse cortical collecting duct cell line, we have studied the physiological control of Na,K-ATPase, the key enzyme that energizes Na reabsorption. Aldosterone, a major regulator of Na transport by the collecting duct, stimulates Na,K-ATPase activity through both recruitment of intracellular pumps and increased total amounts of Na pump subunits. This effect is observed after a lag time of 1 hour and is independent of Na entry through ENaC, but requires de novo transcription and translation. Vasopressin and cAMP, its second messenger, stimulate Na,K-ATPase activity within minutes through translocation of Na pumps from a brefeldin A-sensitive intracellular pool to the plasma membrane. Dysregulation of collecting duct Na,K-ATPase activity is at least in part responsible of the Na retention observed in nephritic syndrome. In this setting, Na,K-ATPase activity and subunit synthesis are specifically increased in CCD. In conclusion, aldosterone, vasopressin, and intracellular Na control the cell surface expression of Na,K-ATPase and translocation from intracellular stores is a major mechanism of regulation of Na,K-ATPase activity in collecting duct principal cells. PMID- 12763892 TI - Themes in ion pump regulation. AB - The Na,K-ATPase is an ion pump present in the plasma membrane. It is of vital importance for cell and body homeostasis and as such is under strict hormonal control. The molecular basis for the acute regulation of Na,K-ATPase is multisite phosphorylation by protein kinases that can alter its behavior. This includes direct effects on the Na,K-ATPase activity, regulation by membrane trafficking, and even dynamic regulation of interaction with regulatory proteins. In shark Na,K-ATPase, the latter includes functional interaction with a small hydrophobic protein of the FXYD protein family, phospholemman-like protein from shark, PLMS. This article summarizes our recent work on the mechanisms involved in the acute regulation of the Na,K-ATPase studied using a plasma membrane preparation from shark salt glands as an epithelial transport model. PMID- 12763893 TI - Isoform-specific regulation of Na+,K+-ATPase endocytosis and recruitment to the plasma membrane. AB - The Na(+),K(+)-ATPase traffics between the plasma membrane and intracellular compartments in response to acute changes in membrane receptor activation. These effects are accomplished by a time-dependent interaction of the Na(+),K(+)-ATPase alpha-subunit with specific intracellular signaling molecules either at the plasma membrane (endocytosis) or at the endosome's membranes (recruitment). Most of these studies have been performed in rat renal epithelial cells in which only the alpha(1) isoenzyme is present. Studies in neurons from the neostriatum in which all three alpha-subunit isoforms are present indicate that neurotransmitter dependent regulation of Na(+),K(+)-ATPase activity displays isoform specificity and also suggest a more complex organization of the intracellular signaling networks controlling Na(+),K(+)-ATPase traffic in mammalian cells. PMID- 12763894 TI - The sodium pump keeps us going. AB - This invited lecture reviews recent evidence that, in skeletal muscle, excitability and contractility depend on the transmembrane distribution of Na(+) and K(+) and the membrane potential, which in turn are determined by the operation of the Na(+)-K(+) pump. Action potentials are elicited by passive fluxes of Na(+) and K(+). Because of their size and sudden onset, these transport events constitute the major challenge for the Na(+)-K(+) pumps. When the Na(+) K(+) pumps cannot readily restore the Na(+)-K(+) gradients, working muscle cells often undergo net loss of K(+) and gain of Na(+). This leads to loss of excitability and force, in particular, in muscles where excitation-induced passive Na(+)-K(+) fluxes are large. Thus, excitability depends on the leak/pump ratio for Na(+) and K(+). When this ratio is increased by inhibition or downregulation of the Na(+)-K(+) pumps, the force decline seen during continued stimulation is accelerated. This effect is highly significant already within the first seconds of electrical stimulation. Fortunately, electrical stimulation also increases Na(+)-K(+) pumping rate within seconds. Thus, maximum increase (20-fold above the resting level) may be reached in 10 seconds, with utilization of all available Na(+)-K(+) pumps. In muscles, where excitability was inhibited by exposure to high [K(+)](o) (10-12.5 mM), activation of the Na(+)-K(+) pumps by hormones or electrical stimulation restored excitability and contractile force. In working muscles, the Na(+)-K(+) pumps, because of rapid activation of their large transport capacity, play a dynamic regulatory role in the second-to-second ongoing restoration and maintenance of excitability and force. The Na(+)-K(+) pumps become a limiting factor for contractile endurance, in particular, if their capacity is reduced by inactivity or disease. PMID- 12763895 TI - Na,K-ATPase and the significance of sodium in the mechanism of potassium-induced relaxation of rat-isolated mesenteric arteries. PMID- 12763896 TI - Molecular activity of sodium pumps in the kidney of mammals and birds. PMID- 12763897 TI - Na+,K+-ATPase subunit isoforms of the developing central nervous system of the lizard Gallotia galloti. PMID- 12763898 TI - Glutamate receptors regulate Na/K-ATPase in cerebellum neurons. PMID- 12763899 TI - Na,K-ATPase isoforms in pregnant and nonpregnant rat uterus. PMID- 12763900 TI - Using Na,K-ATPase itself for large-scale isolation and purification of endogenous digitalis-like factors. PMID- 12763901 TI - Inhibition of the Na,K-ATPase by the antiarrhythmic drug, Bretylium. PMID- 12763902 TI - Muscular K-clearance capacity in vivo must be evaluated on the basis of K and Na,K-ATPase concentrations. PMID- 12763903 TI - Modest K+ restriction provokes insulin resistance of cellular K+ uptake without decrease in plasma K. PMID- 12763904 TI - Na+,K+-ATPase in the marine alga Heterosigma akashiwo. PMID- 12763905 TI - Positive inotropic effect induced by sequence-specific Na+,K+-ATPase antibody in intact cardiac myocytes. PMID- 12763906 TI - Inhibition of purified human kidney Na+,K+-ATPase by cyclosporine A: a possible mechanism for drug human nephrotoxicity. PMID- 12763907 TI - Regulation of Na,K-ATPase by cAMP-dependent protein kinase anchored on membrane via A-kinase anchoring protein subtype, AKAP-150, in rat parotid gland. PMID- 12763908 TI - Porcine kidney extract contains factor(s) that inhibit the ouabain-sensitive isoform of Na,K-ATPase (alpha2) in rat skeletal muscle: a convenient electrophysiological assay. PMID- 12763909 TI - Rat resistance vessels preferentially contain the ouabain-insensitive alpha1 isoform of Na,K-ATPase. PMID- 12763910 TI - Na-pump kinetic properties are differently altered in the brain regions of the cholecystokinin2 receptor-deficient mice. PMID- 12763911 TI - Electrogenic Na+/HCO3- cotransporter rkNBC1 immunolocalized in rat eye. PMID- 12763912 TI - Na,K-ATPase in the regulation of epithelial cell structure. PMID- 12763913 TI - Expression of Na,K-ATPase beta-subunit in transformed MDCK cells increases the translation of the Na,K-ATPase alpha-subunit. PMID- 12763914 TI - Quality control of gastric proton pump in the endoplasmic reticulum by ubiquitin/proteasome system. PMID- 12763915 TI - Mg-ATPase from microsomal fraction of rabbit gastric mucosa is ecto-ATP diphosphohydrolase. PMID- 12763916 TI - Genetics of hypertension: the adducin paradigm. AB - The following were investigated: (1) how we became interested in studying adducin genes and what we know about adducin; (2) studies in animals and humans supporting the role of adducin polymorphisms in hypertension, including some methodological problems related to the dissection of the role of a given genetic molecular mechanism in a complex multifactorial polygenic disease like hypertension; (3) biochemical mechanisms underlying the effect of adducin and its interaction with the Na-K pump; and (4) future directions. PMID- 12763917 TI - Mechanisms of pressure natriuresis: how blood pressure regulates renal sodium transport. AB - An acute increase in blood pressure provokes a rapid decrease in proximal tubule salt and water reabsorption that is central to tubuloglomerular feedback regulation of renal blood flow and glomerular filtration rate and contributes to pressure natriuresis. The molecular mechanisms responsible for this critical homeostatic adjustment were studied. When blood pressure is acutely elevated, apical proximal tubule NHE3 are rapidly redistributed out of the microvilli to intermicrovillar clefts and then endosomal pools, and Na,K-ATPase activity is suppressed. Depressing apical Na(+) entry without hypertension is not sufficient to decrease Na,K-ATPase activity, and depressing Na,K-ATPase activity alone is not sufficient to decrease proximal tubule Na(+) and water reabsorption; thus, it appears that coordinated decreases in both NHE3 surface distribution and Na,K ATPase activity may be important for the response to acute hypertension. Clamping plasma angiotensin II levels blunts the retraction of NHE3 from the cell surface to endosomal pools. The increased volume flow of salt and water to the loop of Henle stimulates Na,K-ATPase activity in this region and provides evidence for a downstream shift in sodium transport during acute hypertension. These same responses in the proximal tubule and loop develop and persist in the spontaneously hypertensive rat. These studies demonstrate that sodium transporters along the nephron are very dynamic, responding quickly to normal fluctuations of blood pressure, and are key to generating the macula densa tubuloglomerular feedback signal and for accommodating increased volume flow through the loop of Henle. PMID- 12763918 TI - Ouabain as a mammalian hormone. AB - Endogenous ouabain changes rapidly in humans and dogs upon physical exercise and is under the control of epinephrine and angiotensin II. Hence, the steroid acts as a rapidly acting hormone. A search for a specific binding globulin for cardiac glycosides in bovine plasma resulted in the identification of the d allotype of the micro chain of IgM whose hydrophobic surfaces interact with cardiotonic steroids and cholesterol. Such IgM complexes might be involved in the hepatic elimination of cardiotonic steroids. Thus, differences in the signaling cascade starting at Na(+),K(+)-ATPase must explain any differences in the action of ouabain and digoxin in the genesis of arterial hypertension. PMID- 12763919 TI - 11-hydroxylation in the biosynthesis of endogenous ouabain: multiple implications. AB - Accumulating evidence indicates that mammals use steroidal glycosides with "digitalis-like" activity. An endogenous ouabain (EO) has been described and is linked with long-term changes in sodium balance and cardiovascular structure and function. In the adrenal gland, the biosynthesis of EO and similar compounds appears to involve cholesterol side-chain cleavage with sequential metabolism of pregnenolone and progesterone. The more distal events in the biosynthesis have not been elucidated. Preliminary work using primary cell cultures from the bovine adrenal cortex suggests that the biosynthesis of EO is affected by inhibitors of 11beta-hydroxylase. Direct participation of 11-hydoxylase in EO synthesis would lead to an 11beta isomer of ouabain in mammals and, in vivo, an 11beta-oriented hydroxyl group would spontaneously form a mixture of two 11-19 hemiketal isomers. The latter isomers would likely be converted back to a single 11beta isomer of ouabain during isolation. The existence of an additional ring in the hemiketals, along with reduced flexion of the steroidal A, B, and C rings, raises the possibility that their in vivo physiological targets and actions differ from the isolated form of EO. PMID- 12763920 TI - Antihypertensive compounds that modulate the Na-K pump. AB - A primary impairment of the kidney sodium excretion has been documented both in hypertensive patients (EH) and genetic animal models (Milan hypertensive rat [MHS]) carrying mutations of the cytoskeletal protein adducin and/or increased plasma levels of endogenous ouabain (EO). Ouabain (OU) itself induces hypertension in rats and both OU and mutated adducin activate the renal Na/K ATPase function both in vivo and in cultured renal cells (NRK). A new antihypertensive agent, PST 2238, able to selectively interact with these alterations has been developed. PST lowers blood pressure (BP) by normalizing the expression and activity of the renal Na-K pump selectively in those rat models carrying the adducin mutation (MHS) and/or increased EO levels (OS) at oral doses of 0.1-10 micro g/kg. In NRK cells either transfected with mutated adducin or incubated with 10(-9) M OU, PST normalizes the Na-K pump activity. Recently, an association between EO and cardiac complications has been observed in both EH and rat models consistent with a prohypertrophic activity of OU. OS rats showed a 10% increase of left ventricle and kidney weights as compared with controls, and PST 2238 (1 micro g/kg OS) prevented both ventricle and renal hypertrophy. This effect was associated with the ability of PST to antagonize the OU-dependent activation of growth-related genes, in the membrane subdomains of caveolae. In conclusion, PST is a new antihypertensive agent that may prevent cardiovascular complications associated with hypertension through the selective modulation of the Na-K pump function. PMID- 12763921 TI - Myocardial Na,K-ATPase and digoxin therapy in human heart failure. AB - The specific binding of digitalis glycosides to the Na,K-ATPase is used as a tool for Na,K-ATPase quantification with high accuracy and precision. In myocardial biopsies from patients with heart failure, total Na,K-ATPase concentration is decreased, and the decrease in Na,K-ATPase concentration correlates with a decrease in heart function. During digitalization, a fraction of remaining pumps are occupied by digoxin. No evidence for an endogenous digitalis-like factor of any clinical importance was obtained. It is recommended that digoxin be administered to heart failure patients who still have dyspnea after institution of mortality-reducing therapy. PMID- 12763922 TI - Expression and cellular localization of Na,K-ATPase isoforms in dog prostate in health and disease. PMID- 12763923 TI - Molecular characterization of a putative sodium/iodide symporter in the South African clawed frog, Xenopus laevis. PMID- 12763924 TI - PSGL-1 participates in E-selectin-mediated progenitor homing to bone marrow: evidence for cooperation between E-selectin ligands and alpha4 integrin. AB - The nature and exact function of selectin ligands involved in hematopoietic progenitor cell (HPC) homing to the bone marrow (BM) are unclear. Using murine progenitor homing assays in lethally irradiated recipients, we found that the P selectin glycoprotein ligand-1 (PSGL-1) plays a partial role in HPC homing to the BM (a reduction of about 35% when the P-selectin binding region is blocked). Blockade of both PSGL-1 and alpha4 integrin did not further enhance the effect of anti-alpha4 integrin (a reduction of about 55%). We suspected that E-selectin ligands might contribute to the remaining homing activity. To test this hypothesis, HPC homing assays were carried out in E-selectin-deficient recipients and revealed a profound alteration in HPC homing when E-selectin and alpha4 integrin were inactivated (> 90% reduction). Competitive assays to test homing of long-term repopulating stem cells revealed a drastic reduction (> 99%) of the homed stem cell activity when both alpha4 integrin and E-selectin functions were absent. Further homing studies with PSGL-1-deficient HPCs pretreated with anti alpha4 integrin antibody revealed that PSGL-1 contributes to approximately 60% of E-selectin ligand-mediated homing activity. Our results thus underscore a major difference between mature myeloid cells and immature stem/progenitor cells in that E-selectin ligands cooperate with alpha4 integrin rather than P-selectin ligands. PMID- 12763925 TI - Oligomerization of RANTES is required for CCR1-mediated arrest but not CCR5 mediated transmigration of leukocytes on inflamed endothelium. AB - Chemokines control inflammatory leukocyte recruitment. The propensity of chemokines such as CC chemokine ligand 5 (CCL5)/RANTES (regulated on activation, normal T cell expressed and secreted) to bind to glycosaminoglycans and to form higher order oligomers has been shown to be essential for its in vivo activity. However, the specific functional relevance of RANTES oligomerization for distinct steps of leukocyte recruitment on inflamed endothelium mediated by the RANTES receptors CC chemokine receptor 1 (CCR1) and CCR5 remains undefined. We studied RANTES mutants with deficient oligomerization in an assay in which recruitment of monocytes and CD45RO+ CD4+ T cells is triggered by RANTES immobilized on activated endothelium under flow conditions. Notably, the formation of higher order RANTES oligomers was crucial for CCR1-mediated arrest but not for CCR5 mediated spreading/transmigration in flow or transendothelial chemotaxis of leukocytes. Efficient leukocyte arrest in flow but not transmigration may thus require the presentation of RANTES oligomers to bridge surface-bound RANTES and CCR1. PMID- 12763926 TI - CD38 is a signaling molecule in B-cell chronic lymphocytic leukemia cells. AB - The prognosis for patients with B-cell chronic lymphocytic leukemia (B-CLL) is generally less favorable for those expressing CD38. Our working hypothesis is that CD38 is not merely a marker in B-CLL, but that it plays a receptor role with pathogenetic potential ruling the proliferation of the malignant clone. CD38 levels were generally low in the patients examined and monoclonal antibody (mAb) ligation was inefficient in signaling. Other cellular models indicated that molecular density and surface organization are critical for CD38 functionality. Interleukin 2 (IL-2) induced a marked up-modulation and surface rearrangement of CD38 in all the patients studied. On reaching a specific expression threshold, CD38 becomes an efficient receptor in purified B-CLL cells. Indeed, mAb ligation is followed by Ca2+ fluxes and by a markedly increased proliferation. The unsuitability of CD38 to perform as a receptor is obviated through close interaction with the B-cell-receptor (BCR) complex and CD19. On mAb binding, CD38 translocates to the membrane lipid microdomains, as shown by a colocalization with the GM1 ganglioside and with CD81, a raft-resident protein. Finally, CD38 signaling in IL-2-treated B-CLL cells prolonged survival and induced the appearance of plasmablasts, providing a pathogenetic hypothesis for the occurrence of Richter syndrome. PMID- 12763927 TI - ALK-positive diffuse large B-cell lymphoma is associated with Clathrin-ALK rearrangements: report of 6 cases. AB - Expression of ALK protein by lymphoid cells and the description of variant anaplastic lymphoma kinase (ALK) translocations have typically been restricted to cases of T-cell and null anaplastic large-cell lymphoma (ALCL). All such cases result from a novel fusion created by the ALK gene on chromosome 2p23 and NPM on 5q35 or other variant translocation partners. A rare variant of diffuse large B cell lymphoma (DLBCL), originally described in 1997, was thought to overexpress full-length ALK in contrast to a chimeric protein characteristic of ALCL. However, full-length ALK protein lacks tyrosine kinase activity and thus the mechanism of oncogenesis has remained elusive. We describe 6 cases of ALK+ DLBCL characterized by a simple or complex t(2;17)(p23;q23) involving the clathrin gene (CLTC) at chromosome band 17q23 and the ALK gene at chromosome band 2p23. All cases were studied using fluorescence in situ hybridization (FISH), complemented in one case with standard cytogenetic analysis, multicolor karyotyping (M-FISH), and reverse transcriptase-polymerase chain reaction. These results clearly demonstrate that most cases of ALK+ DLBCL share the same mechanism of deregulated ALK expression. Moreover, these results demonstrate the presence of CLTC-ALK fusions in these tumors and extend the list of diseases associated with this genetic abnormality to include classical T-cell or null ALCL, ALK+ DLBCL, and inflammatory myofibroblastic tumors. PMID- 12763928 TI - Imatinib mesylate inhibits autonomous erythropoiesis in patients with polycythemia vera in vitro. AB - The overproduction of red blood cells in patients with polycythemia vera (PV) is reflected in vitro by the formation of erythroid burst-forming units (BFU-Es) in the absence of exogenous erythropoietin. In contrast to other myeloproliferative disorders, the molecular mechanism of PV is unknown and no specific chromosomal abnormality has been described. We speculated that imatinib mesylate may reverse the pathological overproduction of red cells by inhibition of autonomous erythropoiesis. In the present study, imatinib mesylate was found to either block or strongly inhibit autonomous BFU-E formation in vitro in all patients tested. Moreover, autonomous BFU-E growth was also markedly reduced by exposure of PV cells to imatinib mesylate prior to cultivation in semisolid medium. The profound effect of imatinib mesylate on autonomous erythropoiesis suggests the involvement of an as yet unidentified kinase in the pathogenesis of PV and should provide the rationale for a forthcoming clinical trial. PMID- 12763929 TI - Fanconi anemia type C-deficient hematopoietic stem/progenitor cells exhibit aberrant cell cycle control. AB - The pathogenesis of bone marrow failure in Fanconi anemia is poorly understood. Suggested mechanisms include enhanced apoptosis secondary to DNA damage and altered inhibitory cytokine signaling. Recent data determined that disrupted cell cycle control of hematopoietic stem and/or progenitor cells disrupts normal hematopoiesis with increased hematopoietic stem cell cycling resulting in diminished function and increased sensitivity to cell cycle-specific apoptotic stimuli. Here, we used Fanconi anemia complementation type C-deficient (Fancc-/-) mice to demonstrate that Fancc-/- phenotypically defined cell populations enriched for hematopoietic stem and progenitor cells exhibit increased cycling. In addition, we established that the defect in cell cycle regulation is not a compensatory mechanism from enhanced apoptosis occurring in vivo. Collectively, these data provide a previously unrecognized phenotype in Fancc-/- hematopoietic stem/progenitor cells, which may contribute to the progressive bone marrow failure in Fanconi anemia. PMID- 12763931 TI - IL-15 drives neonatal T cells to acquire CD56 and become activated effector cells. AB - Expression of one or more natural killer (NK) receptors on T cells may correlate with effector function. This study investigated the frequency of neonatal NK receptor-positive (NKR+) T cells and their expansionary properties with interleukin-2 (IL-2), IL-7, or IL-15. While cord blood contains significantly decreased frequencies of NKR+ T cells compared with adult blood, newborn CD56+CD3+ cells could be expanded 200-fold during culture with IL-15. By depleting CD56+ cells, we were able to determine that this expansion was due to a subpopulation of T cells acquiring CD56 expression. Moreover, CD56 acquisition was associated with a distinct CD8+CD25+ interferon gamma-positive (IFN-gamma+) phenotype. This property could therefore be exploited during bone marrow reconstitution and may partially account for the resilience of the newborn to infection. PMID- 12763930 TI - The third-generation bisphosphonate zoledronate synergistically augments the anti Ph+ leukemia activity of imatinib mesylate. AB - Imatinib mesylate, a competitive inhibitor of Abl tyrosine kinase, is highly effective for the early stages of chronic myelogenous leukemia (CML), but remissions induced in advanced-phase CML and Philadelphia chromosome-positive (Ph+) acute lymphoblastic leukemia tend to be relatively short-lived. Therefore, the search for agents that enhance the anti-Ph+ effect of imatinib mesylate is warranted. We investigated the combined effects of imatinib mesylate and the third-generation bisphosphonate zoledronate (ZOL) on Ph+ leukemias, because ZOL inhibited the prenylation of Ras-related proteins downstream of Bcr/Abl. First, we identified the potency of ZOL in vitro against human leukemic cell lines, including 2 Ph+ and a P-glycoprotein-overexpressing leukemic cell line. ZOL was also effective in vivo because as it prolonged the survival of nonobese diabetic/severe combined immunodeficient (NOD/SCID) mice who were given xenografts with Ph+ BV173 leukemic cells. Next, we showed the in vitro synergistic effects with ZOL and imatinib mesylate for Ph+ cell lines. ZOL combined with imatinib mesylate showed synergistic effects in vivo that prolonged the survival of mice inoculated with BV173. ZOL only minimally inhibited the growth of normal hematopoietic progenitors in vitro, and mice receiving ZOL or imatinib mesylate or both tolerated these treatments well. These findings indicate that ZOL is a potent antileukemic agent that augments synergistically the anti-Ph+ leukemia activity of imatinib mesylate. PMID- 12763932 TI - Phase 1 trial of FVIII gene transfer for severe hemophilia A using a retroviral construct administered by peripheral intravenous infusion. AB - In a phase 1 dose escalation study, 13 subjects with hemophilia A received by peripheral intravenous infusion a retroviral vector carrying a B-domain-deleted human factor VIII (hFVIII) gene. Infusions were well tolerated. Tests for replication competent retrovirus have been negative. Polymerase chain reaction (PCR) analyses demonstrate the persistence of vector gene sequences in peripheral blood mononuclear cells in 3 of 3 subjects tested. Factor VIII was measured in serial samples using both a one-stage clotting assay and a chromogenic assay. While no subject had sustained FVIII increases, 9 subjects had FVIII higher than 1% on at least 2 occasions 5 or more days after infusion of exogenous FVIII, with isolated levels that ranged from 2.3% to 19%. Pharmacokinetic parameters of exogenous FVIII infused into subjects 13 weeks after vector infusion showed an increased half-life (T1/2; P <.02) and area under the curve (AUC, P <.04) compared with prestudy values. Bleeding frequency decreased in 5 subjects compared with historical rates. These results demonstrate that this retroviral vector (hFVIII(V)) is safe and, in some subjects, persists more than a year in peripheral blood mononuclear cells, with measurable factor VIII levels and with increased available FVIII activity (increased T1/2 and AUC) after infusion of exogenous FVIII concentrate. PMID- 12763933 TI - Proliferation kinetics of subpopulations of human marrow cells determined by quantifying in vivo incorporation of [2H2]-glucose into DNA of S-phase cells. AB - This report investigated in vivo turnover kinetics of marrow hematopoietic progenitors and precursors using a recently developed stable isotope-mass spectrometric technique (SIMST). Human subjects were administered a 2-day infusion of 6,6-[2H2]-glucose, a nontoxic stable isotope-labeled form of glucose, which becomes incorporated into DNA of all S-phase cells. The percent [2H2] glucose incorporated into DNA in the form of [2H2]-deoxyadenosine (%[2H2]-dA enrichment) was determined by gas chromatography-mass spectrometry. The rate constant of replacement of unlabeled by labeled DNA strands (labeling kinetics) was used to calculate population turnover kinetics of CD34+ cells, CD133+ cells, and CD133-CD34+ cells. The observed mean replacement half-life (t1/2) was 2.6 days for CD34+ cells, 2.5 days for CD133-CD34+ cells, and 6.2 days for CD133+ cells. Results from the estimated rate constant of replacement of labeled by unlabeled DNA (delabeling kinetics) also demonstrated slower turnover rates for CD133+ cells than for CD133-CD34+ cells. Although there was a relatively rapid initial decrease in the %[2H2]-dA enrichment, low levels of labeled DNA persisted in CD34+ cells for at least 4 weeks. The results indicate the presence of subpopulations of CD34+ cells with relatively rapid turnover rates and subpopulations with a slower t1/2 of 28 days. Results also demonstrate that in vivo [2H2]-glucose-SIMST is sensitive enough to detect differences in turnover kinetics between erythroid and megakaryocyte lineage cells. These studies are the first to demonstrate the use of in vivo [2H2]-glucose-SIMST to measure in vivo turnover kinetics of subpopulations of CD34+ cells and precursors in healthy human subjects. PMID- 12763934 TI - Adoptive transfer of costimulated T cells induces lymphocytosis in patients with relapsed/refractory non-Hodgkin lymphoma following CD34+-selected hematopoietic cell transplantation. AB - We explored the feasibility and toxicity of administering escalating doses of anti-CD3/CD28 ex vivo costimulated T cells as a therapeutic adjunct for patients with relapsed, refractory, or chemotherapy-resistant, aggressive non-Hodgkin lymphoma (NHL) following high-dose chemotherapy and CD34+-selected hematopoietic cell transplantation (HCT). Sixteen patients had infusions on day 14 after HCT of autologous T cells that had been stimulated using beads coated with anti-CD3 and anti-CD28 monoclonal antibodies. At baseline, the subjects had severe quantitative and functional T-cell impairments. The culture procedure partially reversed impaired cytokine responsiveness in T cells in vitro and in vivo. Transient dose-dependent infusion toxicities were observed. There was a rapid reconstitution of lymphocytes; however, there were persistent defects in CD4 T cells. Most interestingly, 5 patients had a delayed lymphocytosis between day 30 and day 120 after HCT. Maximal clinical responses included 5 patients with a complete response (CR), 7 patients with a partial response (PR), and 4 patients with stable disease. At a median follow-up of 33 months (range, 26-60 months), 5 patients are alive with stable or relapsed disease and 3 patients remain in CR. In conclusion, this phase 1 trial demonstrates that adoptive transfer of autologous costimulated T cells (1) is feasible in heavily pretreated patients with advanced NHL, (2) is associated with a rapid recovery of lymphocyte counts, (3) reverses cytokine activation deficits in vitro, and (4) is associated with delayed lymphocytosis in a subset of patients. PMID- 12763936 TI - Induction of pro-angiogenic signaling by a synthetic peptide derived from the second intracellular loop of S1P3 (EDG3). AB - The G-protein-coupled receptors of the endothelial differentiation gene (EDG) family mediate pro-angiogenic activities, such as endothelial cell proliferation, chemotaxis, and vessel morphogenesis. We synthesized and tested the effects of a 9-amino acid peptide (KRX-725), derived from the second intracellular loop of S1P3 (EDG3). KRX-725 mimics the effects of sphingosine 1-phosphate (S1P), the natural ligand of S1P3, by triggering a Gi-dependent MEK-ERK (mitogen-activated protein kinase kinase and extracellular signal-regulated kinase) signal transduction pathway. Using aortic rings as an ex vivo model of angiogenesis, vascular sprouting was assessed in the presence of KRX-725 or S1P. KRX-725 induced extensive and dense vascular sprouts, which contain an elaborated organization of endothelial and smooth muscle layers, including lumen formation. When KRX-725 or S1P was combined with proangiogenic factors, such as basic fibroblast growth factor (bFGF), stem cell factor, or vascular endothelial growth factor, the effect was synergistic, leading to further enhancement of vascular sprouting. KRX-725 also initiated neovascularization in a mouse corneal pocket assay in vivo and showed synergism with bFGF. The specificity of KRX-725 was demonstrated via peptide-induced receptor internalization of S1P3 but not S1P1. The ability of a short peptide to stimulate extensive angiogenesis and to synergize with pro-angiogenic factors suggests that KRX-725 may serve as a useful agent in treating pathologic conditions such as peripheral vascular disease, cardiac ischemia, or tissue grafts. PMID- 12763935 TI - High-dose immunosuppressive therapy and autologous peripheral blood stem cell transplantation for severe multiple sclerosis. AB - There were 26 patients enrolled in a pilot study of high-dose immunosuppressive therapy (HDIT) for severe multiple sclerosis (MS). Median baseline expanded disability status scale (EDSS) was 7.0 (range, 5.0-8.0). HDIT consisted of total body irradiation, cyclophosphamide, and antithymocyte globulin (ATG) and was followed by transplantation of autologous, granulocyte colony-stimulating factor (G-CSF)-mobilized CD34-selected stem cells. Regimen-related toxicities were mild. Because of bladder dysfunction, there were 8 infectious events of the lower urinary tract. One patient died from Epstein-Barr virus (EBV)-related posttransplantation lymphoproliferative disorder (PTLD) associated with a change from horse-derived to rabbit-derived ATG in the HDIT regimen. An engraftment syndrome characterized by noninfectious fever with or without rash developed in 13 of the first 18 patients and was associated in some cases with transient worsening of neurologic symptoms. There were 2 significant adverse neurologic events that occurred, including a flare of MS during mobilization and an episode of irreversible neurologic deterioration after HDIT associated with fever. With a median follow-up of 24 (range, 3-36) months, the Kaplan-Meier estimate of progression (>/= 1.0 point EDSS) at 3 years was 27%. Of 12 patients who had oligoclonal bands in the cerebrospinal fluid at baseline, 9 had persistence after HDIT. After HDIT, 4 patients developed new enhancing lesions on magnetic resonance imaging of the brain. The estimate of survival at 3 years was 91%. Important clinical issues in the use of HDIT and stem cell transplantation for MS were identified; however, modifications of the initial approaches appear to reduce treatment risks. This was a heterogeneous high-risk group, and a phase 3 study is planned to fully assess efficacy. PMID- 12763937 TI - Selective depletion of donor alloreactive T cells without loss of antiviral or antileukemic responses. AB - Poor immune reconstitution after haploidentical stem cell transplantation results in a high mortality from viral infections and relapse. One approach to overcome this problem is to selectively deplete the graft of alloreactive cells using an immunotoxin directed against the activation marker CD25. However, the degree of depletion of alloreactive cells is variable following stimulation with recipient peripheral blood mononuclear cells (PBMCs), and this can result in graft versus host disease (GVHD). We have refined this approach using recipient Epstein-Barr virus (EBV)-transformed lymphoblastoid cell lines (LCLs) as stimulators to activate donor alloreactive T cells. Our studies demonstrate that allodepletion with an anti-CD25 immunotoxin following stimulation with HLA-mismatched host LCLs more consistently depleted in vitro alloreactivity than stimulation with host PBMCs, as assessed in primary mixed lymphocyte reactions (MLRs). Allodepletion using this approach specifically abrogates cytotoxic T-cell responses against host LCLs. In interferon-gamma (IFN-gamma) enzyme-linked immunospot (ELISPOT) assays, antiviral responses to adenovirus and cytomegalovirus (CMV) were preserved following allodepletion. Likewise, using HLA-A2-pp65 tetramers, we have shown that the frequency of CMV-specific T cells is unaffected by allodepletion. Moreover, the donor anti-EBV response is partially retained by recognition of EBV antigens through the nonshared haplotype. Finally, we studied whether allodepletion affects the response to candidate tumor antigens in myeloid malignancies. Using HLA-A2-PR1 tetramer analysis, we found that the frequency of T cells recognizing the PR1 epitope of proteinase 3 was not significantly different in allodepleted and unmanipulated PBMCs from patients with chronic myeloid leukemia (CML) undergoing transplantation. Based on these data, we have embarked on a phase 1 clinical trial of addback of allo-LCL-depleted donor T cells in the haplo-identical setting. PMID- 12763938 TI - Refractory anemia in childhood: a retrospective analysis of 67 patients with particular reference to monosomy 7. AB - Primary myelodysplasia (MDS) without an increased number of blasts is a rare finding in childhood. We performed a retrospective analysis of 67 children with a diagnosis of primary MDS to determine the clinical and hematologic course of the disease. The median age at diagnosis was 8.3 years (range, 0.3-18.1 years). In contrast to refractory anemia in adults, 44% of patients had hemoglobin levels greater than 10 g/100 mL. The median white blood cell count and the absolute neutrophil count were 3.6 x 109/L and 0.9 x 109/L, respectively. Seventy-five percent of patients had thrombocytopenia. Bone marrow was hypocellular in 43% of the patients. Results of cytogenetic analysis showed monosomy 7 in 49%, trisomy 8 in 9%, and other abnormalities in 9% of the patients. The probability of survival 10 years after diagnosis was 0.48 (standard error [SE] = 0.10). Patients with monosomy 7 had significantly higher estimated probabilities of progression to advanced MDS than did patients with other chromosomal anomalies or normal karyotype. Of the 67 children, 41 underwent allogeneic stem cell transplantation (SCT). Patients whose disease did not progress to advanced MDS before SCT had significantly greater probability of survival than patients who experienced progression (0.76 [SE = 0.09] vs 0.36 [SE = 0.16]). SCT improved the outcomes for patients with monosomy 7 and should be offered early in the course of the disease. Recommendations for best treatment options for children with other chromosomal abnormalities or normal karyotype may have to await results of prospective clinical trials. PMID- 12763939 TI - Safety and effectiveness of long-term therapy with the oral iron chelator deferiprone. AB - The identification of a safe, orally active iron chelator is critically important for the prevention of morbidity and early death in patients receiving regular red cell transfusions. Based on our findings in a 1-year multicenter, prospective study of the safety and efficacy of deferiprone in patients with thalassemia major, we have extended the treatment period to 4 years. The mean dose of the chelator was 73 mg/kg per day during 531 patient-years. The rates of agranulocytosis (absolute neutrophil count [ANC] < 500 x 10(9)/L) and milder forms of neutropenia (ANC, 500-1500 x 10(9)/L) were 0.2 and 2.8 per 100 patient years, respectively. Neutropenia occurred significantly more commonly in patients with intact spleens. Gastrointestinal and joint symptoms decreased significantly after the first year of therapy, and led to discontinuation of deferiprone in only one patient in years 2 to 4. The mean alanine aminotransferase (ALT) value of 71 U/L after 4 years of therapy was significantly higher than the baseline value of 61 U/L. Trend analysis showed no increase in the ALT levels or the percentage of patients with ALT levels greater than twice the upper limit of the reference range. Ferritin levels did not change significantly from the values at the time of change from deferoxamine to deferiprone in either the intention-to treat analysis or in the 84 patients who completed 4 years of therapy. Because of concerns regarding the effectiveness of the studied dose of deferiprone, 47 patients discontinued therapy, whereas 15 patients interrupted therapy because of concerns regarding low iron levels. The results of this study help to define the safety and effectiveness of long-term therapy with deferiprone. PMID- 12763940 TI - Inhibition of NF-kappaB by a TAT-NEMO-binding domain peptide accelerates constitutive apoptosis and abrogates LPS-delayed neutrophil apoptosis. AB - Delivery of biologically active peptides into human polymorphonuclear neutrophils (PMNs) has implications for studying cellular functions and may be therapeutically relevant. The transcription factor nuclear factor-kappaB (NF kappaB) regulates the expression of multiple genes controlling inflammation, proliferation, and cell survival. PMNs play a crucial role in first-line defense. Targeting NF-kappaB in these cells may promote apoptosis and therefore facilitate resolution of inflammation. We used an 11-amino acid sequence NEMO-binding domain (NBD) that selectively inhibits the IKKgamma (NEMO)/IKKbeta interaction, preventing NF-kappaB activation. An HIV-TAT sequence served as a highly effective transducing shuttle. We show that lipopolysaccharide (LPS), granulocyte macrophage colony-stimulating factor (GM-CSF), and dexamethasone (DEX) significantly reduced apoptosis after 20 hours. LPS, but not GM-CSF or DEX, activated NF-kappaB as shown by IkappaBalpha degradation, NF-kappaB DNA binding, and transcriptional activity. The TAT-NBD blocked LPS-induced NF-kappaB activation and NF-kappaB-dependent gene expression. TAT-NBD accelerated constitutive PMN apoptosis dose dependently and abrogated LPS-delayed apoptosis. These results provide a proof of principle for peptide delivery by TAT-derived protein transduction domains to specifically inhibit NF-kappaB activity in PMNs. This strategy may help in controlling various cellular functions even in short lived, transfection-resistant primary human cells. PMID- 12763941 TI - Novel pathways of F-actin polymerization in the human neutrophil. AB - Recently we demonstrated the existence of a phosphatidylinositol 3-kinase (PI3K) independent F-actin polymerization during neutrophil pseudopod extension. Here we examine the use of the PI3K-dependent and PI3K-independent pathways of activation by the N-formyl peptide receptor and the chemokine receptors, and the priming of the 2 pathways by granulocyte-macrophage colony-stimulating factor (GM-CSF) and insulin. The inhibition of PI3K activity with wortmannin showed that rate of pseudopod extension stimulated with N-formyl-Met-Leu-Phe (fMLP was mostly dependent on PI3K, while the rate of interleukin-8 (IL-8)-stimulated pseudopod extension was less dependent on PI3K. The incubation of cells with either GM-CSF or insulin increased the rate of pseudopod extension by 50% when the cells were stimulated with IL-8 but not with fMLP. The stimulation with IL-8 phosphorylated the PI3K regulatory subunit. This phosphorylation was enhanced by GM-CSF, which increased PI3K activity and total phosphatidylinositol 3,4,5-trisphosphate (PtdIns(3,4,5)P3) production. The effect of GM-CSF was blocked with wortmannin. In contrast, insulin did not increase p85 phosphorylation and did not enhance PI3K activity or PtdIns(3,4,5)P3 production. The effect of insulin was insensitive to wortmannin; however, it was blocked by an Src homology 2 (SH2) binding peptide. These data indicate that priming of IL-8 activation with GM-CSF was mediated via the PI3Ks of class IA, while priming with insulin used a PI3K independent pathway. PMID- 12763942 TI - Commentary: Diffraction enhanced imaging. PMID- 12763943 TI - Review article: Effects of radiographic contrast media on the lung. AB - The pulmonary adverse effects of intravascular use of water soluble radiographic contrast media (RCM) include bronchospasm, pulmonary oedema and increase in the pulmonary arterial blood pressure (Ppa). Symptomatic bronchospasm is rare but subclinical increase in airways resistance is common after intravascular injection of RCM. Experimental studies have demonstrated that the low osmolar ionic dimer ioxaglate can induce significant bronchospasm in comparison with other types of RCM. Histamine and endothelin, which are potent bronchoconstrictors and released in response to the administration of RCM, do not seem to mediate the bronchospastic effect of RCM. Pretreatment with corticosteroids or antihistamine does not appear to prevent RCM induced bronchospasm, but the administration of beta(2) adrenergic agonist can abolish this adverse effect. RCM induced pulmonary oedema can be secondary to endothelial injury causing an increase in the permeability of the microcirculation. It may also occur in patients with incipient cardiac failure, when large doses of RCM particularly of the high osmolar type are used. A rise in Ppa induced by RCM seems to be secondary to an increase in pulmonary vascular resistance through direct effects on the pulmonary circulation. Low osmolar non ionic monomers induce the least changes in the pulmonary circulation and should be the contrast media of choice for intravascular use in patients with pulmonary hypertension. The mechanisms responsible for the effects of RCM on airway resistance and pulmonary circulation remain unclear. Intrabronchial administration of high osmolar water soluble RCM is dangerous and can induce severe bronchial irritation and pulmonary oedema. Low osmolar RCM are well tolerated by the lungs following aspiration with minimal histological reaction. PMID- 12763944 TI - Early MRI in the management of clinical scaphoid fracture. AB - The incidence of MRI detected scaphoid and other wrist fractures was determined in a clinical setting in patients with suspicion of scaphoid injury and negative initial radiographs. The influence on subsequent patient management was examined. Patients attending Accident and Emergency over a 25 month period with suspected scaphoid fracture and normal scaphoid series plain films were referred for wrist MRI. Scans comprising T(1) weighted spin echo and short tau inversion recovery (STIR) coronal sequences were performed in a dedicated extremity low field MRI scanner within 14 days of injury. Subsequent effects on patient management were ascertained by clinician completed questionnaire. 195 patients were scanned. There were 37 scaphoid fractures (19%), 28 distal radius fractures (14%), 9 fractures of other carpal bones (5%) and 119 studies with no fracture. The management of 180 patients (92%) was altered as a result of the MRI scan. Occult fractures are present in almost two fifths of patients with suspected scaphoid fracture and normal initial plain films. Half of these are scaphoid fractures. MRI allows an early definitive diagnosis to be made, changing patient management in over 90% of cases and should be regarded as the gold standard investigation in this population. PMID- 12763945 TI - X-ray refraction effects: application to the imaging of biological tissues. AB - The purpose of this study was to explore the potential of refraction contrast X ray imaging of biological tissues. Images of dissected mouse lungs, heart, liver and legs were produced using the medical beamline at the Elettra Synchrotron at Trieste, Italy. The technique used was diffraction enhanced imaging. This utilizes a silicon crystal positioned between the tissue sample and the detector to separate refracted X-rays from transmitted and scattered radiation by Bragg diffraction. The contrast in the images produced is related to changes in the X ray refractive index of the tissues, resulting in remarkable clarity compared with conventional X-ray images based on absorption effects. These changes were greatest at the boundaries between different tissues, giving a marked edge enhancement effect and three-dimensional appearance to the images. The technique provides a way of imaging a property of biological tissues not yet exploited, and further studies are planned to identify specific applications in medical imaging. PMID- 12763946 TI - Analysis of benefit:risk ratio and mortality reduction for the UK Breast Screening Programme. AB - A quantitative analysis has been performed to predict the benefit:risk ratio and associated mortality reduction for the UK National Health Service Breast Screening Programme. The analysis is based on the results of an established biological simulation method coupled with dosimetric information and population statistics applicable to the UK breast screening programme. As well as the general breast screening population, the benefit:risk ratios for specific subgroups of women thought to be at higher risk are estimated. The effects of alterations in screening strategy are also investigated. The results indicate favourable benefit:risk ratios and mortality reductions for all women in the programme, with a breast cancer mortality reduction of approximately 9% over the whole UK female population, equivalent to a breast cancer mortality reduction in the region of 25% for the age range 55-69 years. PMID- 12763947 TI - Radiation doses to the legs of radiologists performing interventional procedures: are they a cause for concern? AB - The purpose of this study was to ascertain the magnitude and distribution of doses to the legs of radiologists when performing interventional procedures. LiF:Mg,Ti TLD100 chips were used to measure simultaneously doses to the lower limbs and, for comparison, the hands during 100 interventional procedures. Results show leg dose was dependent upon type and complexity of procedure, equipment used and whether lead protection was available. Where no lead protection was used, the doses to the lower limbs were frequently similar to or higher than those received by the hands. The mean dose to the legs ranged from 0.19 mSv to 2.61 mSv per procedure, compared with 0.04 mSv to 1.25 mSv to the hands. During transjugular intrahepatic portosystemic shunt and embolisation procedures the leg dose could be as much as 2-3 times greater than that to the hands. When lead protection was used, the dose to the legs was reduced significantly to 0.02 mSv to 0.5 mSv per procedure. A clear linear relationship was shown between the dose-area product (DAP) reading and the dose to the feet of the radiologist. As a "rule of thumb", a DAP reading of 100 Gy cm(2) will give a dose of 1 mSv to the legs, if no lead protection was used, dropping to approximately 0.02 mSv if lead protection was present. This study demonstrates that the dose to the legs of radiologists can be higher than that to the hands when no lead protection is used. The inclusion of a lead screen to protect the legs is an effective method of dose reduction when performing interventional procedures. PMID- 12763948 TI - Interexamination variation of whole breast ultrasound. AB - The purpose of this study was to determine the interexamination agreement of ultrasound of the breasts. This includes the complete process of performing whole breast ultrasound and interpreting the dynamic scanning and the static images by one person. In a prospective study, 58 patients with a clinical indication for mammography underwent an ultrasound examination of both breasts by three independent sonographers. The sonographers had full knowledge of the physical and mammographic findings. Histology and 12 month follow-up were used as the reference standard. Interobserver variability for both mammography and breast ultrasound was measured using linearly weighted kappa statistics. Receiver operator characteristic curves were constructed to compare the diagnostic performance of the observers. The interexamination agreement for the score of the probability of malignancy after mammography was substantial (kappas ranged from 0.63 to 0.65). The interexamination agreement for the final score of the probability of malignancy after mammography and ultrasound examination was slightly better (kappas ranged from 0.72 to 0.75). The area under the receiver operating characteristic curves after mammography and ultrasound examination ranged from 0.97 to 0.98. Ultrasound examination of the whole breast shows a substantial interexamination agreement. Ultrasound examination of the breast adds consistency to mammography and physical examination. PMID- 12763949 TI - Short communication: Practical aspects for the evaluation of skin doses in interventional cardiology using a new slow film. AB - Mapping skin doses in complex fluoroscopy interventions is useful to determine the probability of a possible injury, to detect areas of overlapping irradiation fields and to obtain a permanent register of the most exposed patient skin areas. To fulfil this task, large films with slow X-ray response can be used. Recently, Kodak has introduced a new radiotherapy verification film, named EDR2 (Extended Dose Range). The aim of this paper is to analyse the possibilities of using this new film for estimating skin dose distributions in interventions with potentially higher doses, such as complex percutaneous transluminal coronary angioplasty (PTCA), intravascular brachytherapy procedures (IVB) or cardiac ablations. The EDR2 film by Kodak is an improved option to be used in interventional cardiology to obtain maps of patient skin doses and to estimate maximum skin doses up to 1400 mGy. Film kVp dependence is negligible and the processor conditions can be standardized to obtain skin dose estimations. The linear range for accurate dose measurements is from 50 mGy to 500 mGy. PMID- 12763950 TI - Case report: Unusual peritoneal spreading by metastatic osteosarcoma of the tibia. AB - We report a 17-year-old boy with chondroblastic osteosarcoma initially found in left proximal tibia. He received surgical resection and chemotherapy. However, a lung metastasis was found 4 years later. Despite intensive chemotherapy, the metastatic osteosarcoma of lung continued to invade the ribs and later into retroperitoneum and liver. The metastatic pattern of chondroblastic osteosarcoma of tibia directly to the chest and then into the abdomen is unusual. PMID- 12763951 TI - Case report: Mucoepidermoid carcinoma in a patient with congenital agenesis of the left upper lobe. AB - This report describes an unusual case of a mucoepidermoid carcinoma developing in a patient with congenital left upper lobe agenesis. Mucoepidermoid tumours most often develop in major bronchi and present as lobar collapse, post-obstructive pneumonia or as a mass lesion on chest radiography. On CT, the tumour is smooth, well defined, homogeneous and may enhance with intravenous contrast. Tumour calcification has been reported. Lobar agenesis often presents as a co-incidental finding on chest radiography as total or almost complete absence of aeration of the affected lung. Thoracic CT confirms the presence of the underdeveloped lung, pulmonary artery and bronchus and associated mediastinal shift and herniation of the contralateral lung. A literature review of these two conditions is presented. PMID- 12763952 TI - Case report: Squamous carcinoma in an oesophageal foregut cyst. AB - Cysts within the oesophageal wall may represent inclusion cysts, retention cysts or developmental cysts. Foregut duplications are developmental anomalies, which occur as a result of abnormal canalization of the foregut during intrauterine life. Malignant transformation is an extremely rare event occurring within oesophageal cysts, adenocarcinoma being the most common histology. We report a case of squamous cell carcinoma arising within an oesophageal cyst affecting the upper third of the oesophagus. The malignant cyst was not amenable to primary surgical resection and hence was treated using chemo-radiotherapy. The treatment gave good disease control, at the expense of a high oesophageal stricture. Chemo radiotherapy is an alternative treatment modality to achieve long-term disease control in squamous cell carcinoma complicating oesophageal foregut cyst when primary surgical resection is not possible. PMID- 12763953 TI - Pictorial review: MRI of chronic spinal cord injury. AB - Patients who have suffered a spinal cord injury and who demonstrate new or changing clinical features such as increasing myelopathy, ascending neurological level, pain or increasing muscle spasms may have developed a late complication such as post-traumatic syrinx. MRI is the investigation of choice for assessment of chronic spinal cord injury. The aim of this pictorial review is to illustrate the various late appearances of the injured spinal cord. PMID- 12763954 TI - Case of the month. A lump in the sole of the foot. PMID- 12763955 TI - The failures of two contracts. PMID- 12763956 TI - Patients and the new contracts. PMID- 12763957 TI - New leader, new hope for WHO. PMID- 12763958 TI - Why clinical information standards matter. PMID- 12763959 TI - United Kingdom has its first confirmed case of SARS. PMID- 12763960 TI - US and Germany give late support to WHO tobacco accord. PMID- 12763961 TI - US guidelines say blood pressure of 120/80 mm Hg is not "normal". PMID- 12763970 TI - Drug company accused of breaking advertising regulations. PMID- 12763972 TI - Media misled the public over the MMR vaccine, study says. PMID- 12763973 TI - Poor security is biggest impediment to health care in Iraq. PMID- 12763974 TI - Patient confidentiality challenged over HIV test results. PMID- 12763975 TI - Law against asylum seekers may have public health impact. PMID- 12763976 TI - Chinese scientists must test wild animals to find the host of SARS. PMID- 12763977 TI - Identifying the fallen. PMID- 12763979 TI - Quality improvement forum sets sights on developing countries and on patients. PMID- 12763980 TI - Beta lactam monotherapy versus beta lactam-aminoglycoside combination therapy for fever with neutropenia: systematic review and meta-analysis. AB - OBJECTIVE: To compare the effectiveness of beta lactam monotherapy versus beta lactam-aminoglycoside combination therapy in the treatment of patients with fever and neutropenia. DATA SOURCES: Medline, Embase, Lilacs, the Cochrane Library, and conference proceedings to 2002. References of included studies and contact with authors. No restrictions on language, year of publication, or publication status. STUDY SELECTION: All randomised trials of beta lactam monotherapy compared with beta lactam-minoglycoside combination therapy as empirical treatment for patients with fever and neutropenia. DATA SELECTION: Two reviewers independently applied selection criteria, performed quality assessment, and extracted data. An intention to treat approach was used. Relative risks were pooled with the random effect model. MAIN OUTCOME MEASURE: All cause fatality. RESULTS: Forty seven trials with 7807 patients met inclusion criteria. Nine trials compared the same beta lactam. There was no significant difference in all cause fatality (relative risk 0.85, 95% confidence interval 0.72 to 1.02). For success of treatment there was a significant advantage with monotherapy (0.92, 0.85 to 0.99), though there was considerable heterogeneity among trials. There was no significant difference between monotherapy and combination treatment in trials that compared the same beta lactam, whereas there was major advantage with monotherapy in trials that compared different beta lactams (0.87, 0.80 to 0.93). Rates of superinfection were similar. Adverse events, including those associated with severe morbidity, were significantly more common in the combination treatment group. Detected flaws in methods did not affect results. CONCLUSIONS: For patients with fever and neutropenia there is no clinical advantage in treatment with beta lactam aminoglycoside combination therapy. Broad spectrum beta lactams as monotherapy should be regarded as the standard of care for such patients. PMID- 12763981 TI - Stepping down inhaled corticosteroids in asthma: randomised controlled trial. AB - OBJECTIVES: To determine whether the dose of inhaled corticosteroids can be stepped down in patients with chronic stable asthma while maintaining control. DESIGN: One year, randomised controlled, double blind, parallel group trial. SETTING: General practices throughout western and central Scotland. PARTICIPANTS: 259 adult patients with asthma receiving regular treatment with inhaled corticosteroids at high dose (mean dose 1430 microg beclomethasone dipropionate). INTERVENTIONS: Participants were allocated to receive either no alteration to their dose of inhaled corticosteroid (control) or a 50% reduction in their dose if they met criteria for stable asthma (stepdown). MAIN OUTCOME MEASURES: Comparison of asthma exacerbation rates, asthma related visits to general practice and hospital, health status measures, and corticosteroid dosage between the two groups. RESULTS: The proportions of subjects with asthma exacerbations were not significantly different (stepdown 31%, control 26%, P=0.354). Similarly, the numbers of visits to general practice or hospital and the disease specific and generic measures of health status over the one year period were not significantly different. On average the stepdown group received 348 microg (95% confidence interval 202 microg to 494 microg) of beclomethasone dipropionate less per day than the controls (a difference of 25%), with no difference in the annual dose of oral corticosteroids between the two treatment regimens. CONCLUSIONS: By adopting a stepdown approach to the use of inhaled steroids at high doses in asthma a reduction in the dose can be achieved without compromising asthma control. PMID- 12763983 TI - Evaluation of suicide rates in rural India using verbal autopsies, 1994-9. PMID- 12763982 TI - Empirical prescribing for dyspepsia: randomised controlled trial of test and treat versus omeprazole treatment. AB - OBJECTIVE: To compare the efficacy of a "Helicobacter pylori test and treat" strategy with that of an empirical trial of omeprazole in the non-endoscopic management by empirical prescribing of young patients with dyspepsia. DESIGN: Randomised controlled trial. SETTING: Hospital gastroenterology unit. PARTICIPANTS: 219 patients under 45 years old presenting with dyspepsia without alarm symptoms. INTERVENTION: Patients received treatment with omeprazole 20 mg (group A) or with a urea breath test followed by an eradication treatment in case of H pylori infection or omeprazole alone in non-infected patients (group B). Lack of improvement or recurrence of symptoms prompted endoscopy. MAIN OUTCOME MEASURES: Improvement in symptoms assessed by a dyspepsia severity score every two months; use of medical resources (endoscopic workload and medical consultation); clinical outcome. RESULTS: 96/109 (88%) patients in group A and 61/110 (55%) in group B (P < 0.0001) had endoscopy: in 19 patients in group A and 32 in group B (20/67 infected and 12/43 non-infected) because of no improvement; in 77 further patients in group A and 29 in group B (7 infected and 22 non infected) because of recurrence of symptoms during follow up. Endoscopy showed peptic ulcers only in group A; oesophagitis occurred significantly more often in group B than in group A. About 80% of examinations were normal in both groups, but nine duodenal scars occurred in group A. CONCLUSIONS: Eradication treatment allows resolution of symptoms in a large number of patients with dyspepsia and reduces the endoscopic workload. After a trial of omeprazole, symptoms recur in nearly every patient. Such treatment is also likely to mask an appreciable number of peptic ulcers and cases of oesophagitis. PMID- 12763984 TI - Participation in screening for cardiovascular risk by people with schizophrenia or similar mental illnesses: cross sectional study in general practice. PMID- 12763985 TI - Iron supplementation for unexplained fatigue in non-anaemic women: double blind randomised placebo controlled trial. AB - OBJECTIVE: To determine the subjective response to iron therapy in non-anaemic women with unexplained fatigue. DESIGN: Double blind randomised placebo controlled trial. SETTING: Academic primary care centre and eight general practices in western Switzerland. PARTICIPANTS: 144 women aged 18 to 55, assigned to either oral ferrous sulphate (80 mg/day of elemental iron daily; n=75) or placebo (n=69) for four weeks. MAIN OUTCOME MEASURES: Level of fatigue, measured by a 10 point visual analogue scale. RESULTS: 136 (94%) women completed the study. Most had a low serum ferritin concentration; 5000 samples for the presence of antineuronal antibodies. A total of 137 patients were identified with a paraneoplastic neurological syndrome and high titre (> or =400) antineuronal antibodies. Fifty (36%) of these patients had antibody-associated PCD, including 19 anti-Yo, 16 anti-Hu, seven anti-Tr, six anti-Ri and two anti-mGluR1. Because of the low number, the anti-mGluR1 patients were excluded from the statistical analysis. While 100% of patients with anti-Yo, anti-Tr and anti-mGluR1 antibodies suffered PCD, 86% of anti-Ri and only 18% of anti-Hu patients had PCD. All patients presented with subacute cerebellar ataxia progressive over weeks to months and stabilized within 6 months. The majority of patients in all antibody groups had both truncal and appendicular ataxia. The frequency of nystagmus and dysarthria was lower in anti-Ri patients (33 and 0%). Later in the course of the disease, involvement of non-cerebellar structures occurred most frequently in anti-Hu patients (94%). In 42 patients (84%), a tumour was detected. The most commonly associated tumours were gynaecological and breast cancer (anti-Yo and anti-Ri), lung cancer (anti-Hu) and Hodgkin's lymphoma (anti-Tr and anti-mGluR1). In one anti-Hu patient, a suspect lung lesion on CT scan disappeared while the PCD evolved. Seven patients improved by at least 1 point on the Rankin scale, while 16 remained stable and 27 deteriorated. All seven patients that improved received antitumour treatment for their underlying cancer, resulting in complete remission. The functional outcome was best in the anti-Ri patients, with three out of six improving neurologically and five were able to walk at the time of last follow-up or death. Only four out of 19 anti-Yo and four out of 16 anti-Hu patients remained ambulatory. Also, survival from time of diagnosis was significantly worse in the anti-Yo (median 13 months) and anti-Hu (median 7 months) patients compared with anti-Tr (median >113 months) and anti-Ri (median >69 months). Patients receiving antitumour treatment (with or without immunosuppressive therapy) lived significantly longer [hazard ratio (HR) 0.3; 95% confidence interval (CI) 0.1-0.6; P = 0.004]. Patients > or =60 years old lived somewhat shorter from time of diagnosis, although statistically not significant (HR 2.9; CI 1.0-8.5; P = 0.06). PMID- 12764062 TI - Expression profiling identifies responder and non-responder phenotypes to interferon-beta in multiple sclerosis. AB - Autoimmune diseases such as multiple sclerosis are characterized by complex genetic traits and pathomechanisms that translate into clinical heterogeneity. This wide heterogeneity of multiple sclerosis as well as different biological responses to immunomodulatory drugs can be expected to contribute to differential treatment responses. Strategies that dissect the relationship between the treatment response and the biological characteristics in individual patients are valuable not only as a clinical tool, but also in leading to a better understanding of the disease. Here we address the in vitro and ex vivo RNA expression profile under one approved therapy of multiple sclerosis, interferon beta (IFN-beta, Betaseron), by cDNA microarrays and demonstrate that non responder and responder phenotypes to IFN-beta as assessed by longitudinal gadolinium-enhanced MRI scans and clinical disease activity differ in their ex vivo gene expression profile. These findings will help to better elucidate the mechanism of action of IFN-beta in relation to different disease patterns and eventually lead to optimized therapy. PMID- 12764063 TI - Neural correlates of outcome after stroke: a cross-sectional fMRI study. AB - Recovery of motor function after stroke may occur over weeks or months and is often attributed to neuronal reorganization. Functional imaging studies investigating patients who have made a good recovery after stroke have suggested that recruitment of other motor-related networks underlies this recovery. However, patients with less complete recovery have rarely been studied, or else the degree of recovery has not been taken into account. We set out to investigate the relationship between the degree of recovery after stroke and the pattern of recruitment of brain regions during a motor task as measured using functional MRI. We recruited 20 patients who were at least 3 months after their first ever stroke, and 26 right-handed age-matched control subjects. None of our patients had infarcts involving the hand region of the primary motor cortex. All subjects were scanned whilst performing an isometric, dynamic visually paced handgrip task. The degree of functional recovery of each patient was assessed using a battery of outcome measures. Single-patient versus control group analysis revealed that patients with poor recovery were more likely to recruit a number of motor-related brain regions over and above those seen in the control group during the motor task, whereas patients with more complete recovery were more likely to have 'normal' task-related brain activation. Across the whole patient group and across stroke subtypes, we were able to demonstrate a negative correlation between outcome and the degree of task-related activation in regions such as the supplementary motor area, cingulate motor areas, premotor cortex, posterior parietal cortex, and cerebellum. This negative correlation was also seen in parts of both contralateral and ipsilateral primary motor cortex. These results further our understanding of the recovery process by demonstrating for the first time a clear relationship between task-related activation of the motor system and outcome after stroke. PMID- 12764064 TI - Epileptic fast intracerebral EEG activity: evidence for spatial decorrelation at seizure onset. AB - Low-voltage rapid discharges (or fast EEG ictal activity) constitute a characteristic electrophysiological pattern in focal seizures of human epilepsy. They are characterized by a decrease of signal voltage with a marked increase of signal frequency (typically beyond 25 Hz). They have long been observed in stereoelectroencephalographic (SEEG) signals recorded with intra-cerebral electrodes, generally occurring at seizure onset and simultaneously involving distinct brain regions. Spectral properties of rapid ictal discharges as well as spatial correlations measured between SEEG signals generated from distant sites before, during and after these discharges were studied. Cross-correlation estimates within typical EEG sub-bands and statistical tests performed in 10 patients suffering from partial epilepsy (frontal, temporal or fronto-temporal) reveal that SEEG signals are significantly de-correlated during the discharge period compared with periods that precede and follow this discharge. These results can be interpreted as a functional decoupling of distant brain sites at seizure onset followed by an abnormally high re-coupling when the seizure develops. They lead to the concept of 'disruption' that is complementary of that of 'activation' (revealed by significantly high correlations between signals recorded during seizures), both giving insights into our understanding of pathophysiological processes involved in human partial epilepsies as well as in the interpretation of clinical semiology. PMID- 12764065 TI - Decisional role of the dorsolateral prefrontal cortex in ocular motor behaviour. AB - Three patients with a unilateral cortical lesion affecting the dorsolateral prefrontal cortex (DLPFC), i.e. Brodmann area 46, were tested using different paradigms of reflexive saccades (gap and overlap tasks), intentional saccades (antisaccades, memory-guided and predictive saccades) and smooth pursuit movements. Visually guided saccades with gap and overlap, latency of correct antisaccades and memory-guided saccades and the gain of smooth pursuit were normal, compared with controls. These results confirm our anatomical data showing that the adjacent frontal eye field (FEF) was unimpaired in these patients. The specific pattern of abnormalities after a unilateral DLPFC lesion, compared with that of the FEF lesions previously reported, consists mainly of: (i) a bilateral increase in the percentage of errors in the antisaccade task (misdirected reflexive saccades); (ii) a bilateral increase in the variable error in amplitude, without significant decrease in the gain, in the memory-guided saccade task; and (iii) a bilateral decrease in the percentage of anticipatory saccades in the predictive task. Taken together, these results suggest that the DLPFC plays a crucial role in the decisional processes, preparing saccades by inhibiting unwanted reflexive saccades (inhibition), maintaining memorized information for ongoing intentional saccades (short-term spatial memory) or facilitating anticipatory saccades (prediction), depending upon current external environmental and internal circumstances. PMID- 12764066 TI - Specific brain processing of facial expressions in people with alexithymia: an H2 15O-PET study. AB - Alexithymia is a personal trait characterized by a reduced ability to identify and describe one's own feelings and is known to contribute to a variety of physical and behavioural disorders. To elucidate the pathogenesis of stress related disorders and the normal functions of emotion, it is important to investigate the neurobiology of alexithymia. Although several neurological models of alexithymia have been proposed, there is very little direct evidence for the neural correlates of alexithymia. Using PET, we studied brain activity in subjects with alexithymia when viewing a range of emotional face expressions. Twelve alexithymic and 12 non-alexithymic volunteers (all right-handed males) were selected from 247 applicants on the basis of the 20-item Toronto Alexithymia Scale (TAS-20). Regional cerebral blood flow (rCBF) was measured with H(2)(15)O PET while the subjects looked at angry, sad and happy faces with varying emotional intensity, as well as neutral faces. Brain response in the subjects with alexithymia significantly differed from that in the subjects without alexithymia. The alexithymics exhibited lower rCBF in the inferior and middle frontal cortex, orbitofrontal cortex, inferior parietal cortex and occipital cortex in the right hemisphere than the non-alexithymics. Additionally, the alexithymics showed higher rCBF in the superior frontal cortex, inferior parietal cortex and cerebellum in the left hemisphere when compared with the non alexithymics. A covariance analysis revealed that rCBF in the inferior and superior frontal cortex, orbitofrontal cortex and parietal cortex in the right hemisphere correlated negatively with individual TAS-20 scores when viewing angry and sad facial expressions, and that no rCBF correlated positively with TAS-20 scores. Moreover, the anterior cingulate cortex and insula were less activated in the alexithymics' response to angry faces than their response to neutral faces. These results suggest that people with alexithymia process facial expressions differently from people without alexithymia, and that this difference may account for the disorder of affect regulation and consequent peculiar behaviour in people with alexithymia. PMID- 12764067 TI - Autosomal dominant restless legs syndrome maps on chromosome 14q. AB - Restless legs syndrome (RLS) is a common neurological disorder characterized by an irresistible desire to move the extremities associated with paraesthesia/dysaesthesia. These symptoms occur predominantly at rest and worsen at night, resulting in nocturnal insomnia and chronic sleep deprivation. In this paper, we show significant evidence of linkage to a new locus for RLS on chromosome 14q13-21 region in a 30-member, three-generation Italian family affected by RLS and periodic leg movements in sleep (PLMS). This is the second RLS locus identified so far and the first consistent with an autosomal dominant inheritance pattern. The new RLS critical region spans 9.1 cM, between markers D14S70 and D14S1068. The maximum two-point log of odds ratio score value, of 3.23 at theta = 0.0, was obtained for marker D14S288. The accurate clinical evaluation of RLS-affected, as well as unaffected, family members allowed for the configuring of RLS as a phenotypic spectrum ranging from PLMS to RLS. Motor component, both while awake and during sleep, was an important aspect of the phenotype in the family analysed. The complementary clinical and genetic studies on multiplex families are likely to be of the utmost importance in unfolding the complete expressivity of RLS phenotype spectrum. PMID- 12764068 TI - California Verbal Learning Test: performance by patients with focal frontal and non-frontal lesions. AB - Although frontal lobe lesions do not cause classic amnesia, they may disrupt learning and memory in a number of ways. To investigate in finer detail the regions of frontal injury that are associated with impaired learning and to define the cognitive processing deficits specific to each region that disrupt memory, we compared 33 patients with focal frontal injury with patients with non frontal injury and with normal controls on a standard neuropsychological instrument, the California Verbal Learning Test (CVLT). Subgroups of patients with distinct lesion site profiles were compared in a number of learning measures. All of the subgroups of patients with frontal lesions (with one exception) had inefficient learning due to poor implementation of a strategy of subjective organization. Despite this organizational deficiency, the performance of patients with frontopolar lesions normalized across trials. Only the subgroups with lesions centred either on the left posterior dorsolateral frontal region or the posterior medial frontal region had overall impaired learning and recall. The left posterior dorsolateral frontal group was most significantly impaired on all measures. This recall impairment was secondary to a mild lexical-semantic deficit. A recognition memory deficit in the same group was due to an abnormal response bias. Several groups had a modest increase in perseverative recalls; the underlying mechanisms differed. Disruption of different cognitive processes associated with specific frontal regions underlies the varied patterns of memory impairment. This study has demonstrated even finer differentiations within the frontal region than previously known. PMID- 12764069 TI - Glatiramer acetate treatment in Devic's neuromyelitis optica. PMID- 12764071 TI - TGF-beta signaling and its functional significance in regulating the fate of cranial neural crest cells. AB - Members of the transforming growth factor-beta (TGF-beta) superfamily regulate cell proliferation, differentiation, and apoptosis, and control the development and maintenance of most tissues. TGF-beta signal is transmitted through the phosphorylation of Smad proteins by TGF-beta receptor serine/threonine kinase. During craniofacial development, TGF-beta may regulate the fate specification of cranial neural crest cells. These cells are multipotent progenitors and capable of producing diverse cell types upon differentiation. Here we summarize evidence that TGF-beta ligands and their signaling intermediates have significant roles in patterning and specification of cranial neural crest cells. The biological function of TGF-beta is carried out through the regulation of transcriptional factors during embryogenesis. PMID- 12764072 TI - Glucan-binding proteins of the oral streptococci. AB - The synthesis of extracellular glucan is an integral component of the sucrose dependent colonization of tooth surfaces by species of the mutans streptococci. In investigators' attempts to understand the mechanisms of plaque biofilm development, several glucan-binding proteins (GBPs) have been discovered. Some of these, the glucosyltransferases, catalyze the synthesis of glucan, whereas others, designated only as glucan-binding proteins, have affinities for different forms of glucan and contribute to aspects of the biology of their host organisms. The functions of these latter glucan-binding proteins include dextran-dependent aggregation, dextranase inhibition, plaque cohesion, and perhaps cell wall synthesis. In some instances, their glucan-binding domains share common features, whereas in others the mechanism for glucan binding remains unknown. Recent studies indicate that at least some of the glucan-binding proteins modulate virulence and some can act as protective immunogens within animal models. Overall, the multiplicity of GBPs and their aforementioned properties are testimonies to their importance. Future studies will greatly advance the understanding of the distribution, function, and regulation of the GBPs and place into perspective the facets of their contributions to the biology of the oral streptococci. PMID- 12764073 TI - The biochemistry and physiology of metallic fluoride: action, mechanism, and implications. AB - Fluoride is a well-known G protein activator. Activation of heterotrimeric GTP binding proteins by fluoride requires trace amounts of Al3+ or Be2+ ions. AlFx mimics a gamma-phosphate at its transition state in a Galpha protein and is therefore able to inhibit its GTPase activity. AlFx also forms complexes with small GTP-binding proteins in the presence of their GTPase-activating proteins (GAP). As phosphate analogs, AlFx or BeFx affect the activity of a variety of phosphoryl transfer enzymes. Most of these enzymes are fundamentally important in cell signal transduction or energy metabolism. Al3+ and F- tend to form stable complexes in aqueous solution. The exact structure and concentration of AlFx depend on the pH and the amount of F- and Al3+ in the solution. Humans are exposed to both F and Al. It is possible that Al-F complexes may be formed in vivo, or formed in vitro prior to their intake by humans. Al-F complexes may play physiological or pathological roles in bone biology, fluorosis, neurotoxicity, and oral diseases such as dental caries and periodontal disease. The aim of this review is to discuss the basic chemical, biochemical, and toxicological properties of metallic fluoride, to explore its potential physiological and clinical implications. PMID- 12764074 TI - Oral diseases possibly associated with hepatitis C virus. AB - Morbidity associated with hepatitis C virus (HCV) infection can involve a variety of extrahepatic conditions, including lichen planus (LP) and sialadenitis, predominantly or exclusively involving the oral region, conditions which have been largely neglected in reviews. The literature suggests that HCV-infected patients may frequently have Sjogren-like sialadenitis with mild clinical symptoms, whereas oral LP may be significantly associated with HCV infections in Southern Europe and Japan but not in Northern Europe. These geographical differences could be related to immunogenetic factors such as the HLA-DR6 allele, significantly expressed in Italian patients with OLP and HCV. Analysis of experimental data suggests that HCV could be involved in the pathogenesis of both these diseases. Moreover, parotid lymphoma may arise in patients with sialadenitis, mainly with type II cryoglobulinemia. Little attention has been paid to oral health needs in HCV-infected patients and the variable effect of interferon-alpha therapy on oral tissues. Further research is needed, because of the potentially great influence of oral diseases possibly linked to HCV on the quality of life of millions of patients. PMID- 12764075 TI - Yeasts in apical periodontitis. AB - Microbiological reports of apical periodontitis have revealed that yeasts can be isolated from approximately 5-20% of infected root canals. They occur either in pure cultures or together with bacteria. Almost all isolated yeasts belong to the genus Candida, and the predominant species is C. albicans. Pheno- and genotypic profiles of C. albicans isolates show heterogeneity comparable with those of isolates from other oral sites. C. albicans expresses several virulence factors that are capable of infecting the dentin-pulp complex, including dentinal tubules. This causes, consequentially, an inflammatory response around the root apex, which suggests a pathogenic role for this organism in apical periodontitis. Yeasts are particularly associated with persistent root canal infections that do not respond favorably to conservative root canal therapy. This may be due to the resistance of all oral Candida species against a commonly used topical medicament, calcium hydroxide. However, other antimicrobial agents may offer alternative therapeutic approaches and improve the treatment of these persistent cases of apical periodontitis. PMID- 12764076 TI - Biomechanical behavior of the temporomandibular joint disc. AB - The temporomandibular joint (TMJ) disc consists mainly of collagen fibers and proteoglycans constrained in the interstices of the collagen fiber mesh. This construction results in a viscoelastic response of the disc to loading and enables the disc to play an important role as a stress absorber during function. The viscoelastic properties depend on the direction (tension, compression, and shear) and the type of the applied loading (static and dynamic). The compressive elastic modulus of the disc is smaller than its tensile one because the elasticity of the disc is more dependent on the collagen fibers than on the proteoglycans. When dynamic loading occurs, the disc is likely to behave less stiffly than under static loading because of the difference of fluid flow through and out of the disc during loading. In addition, the mechanical properties change as a result of various intrinsic and extrinsic factors in life such as aging, trauma, and pathology. Information about the viscoelastic behavior of the disc is required for its function to be understood and, for instance, for a suitable TMJ replacement device to be constructed. In this review, the biomechanical behavior of the disc in response to different loading conditions is discussed. PMID- 12764077 TI - Differential control of adrenal and sympathetic catecholamine release by alpha 2 adrenoceptor subtypes. AB - In the adrenergic system, release of the neurotransmitter norepinephrine from sympathetic nerves is regulated by presynaptic inhibitory alpha2-adrenoceptors, but it is unknown whether release of epinephrine from the adrenal gland is controlled by a similar short feedback loop. Using gene-targeted mice we demonstrate that two distinct subtypes of alpha2-adrenoceptors control release of catecholamines from sympathetic nerves (alpha 2A) and from the adrenal medulla (alpha 2C). In isolated mouse chromaffin cells, alpha2-receptor activation inhibited the electrically stimulated increase in cell capacitance (a correlate of exocytosis), voltage-activated Ca2+ current, as well as secretion of epinephrine and norepinephrine. The inhibitory effects of alpha2-agonists on cell capacitance, voltage-activated Ca2+ currents, and on catecholamine secretion were completely abolished in chromaffin cells isolated from alpha 2C-receptor deficient mice. In vivo, deletion of sympathetic or adrenal feedback control led to increased plasma and urine norepinephrine (alpha 2A-knockout) and epinephrine levels (alpha 2C-knockout), respectively. Loss of feedback inhibition was compensated by increased tyrosine hydroxylase activity, as detected by elevated tissue dihydroxyphenylalanine levels. Thus, receptor subtype diversity in the adrenergic system has emerged to selectively control sympathetic and adrenal catecholamine secretion via distinct alpha2-adrenoceptor subtypes. Short-loop feedback inhibition of epinephrine release from the adrenal gland may represent a novel therapeutic target for diseases that arise from enhanced adrenergic stimulation. PMID- 12764078 TI - Glucocorticoids regulate the expression of the mouse urocortin II gene: a putative connection between the corticotropin-releasing factor receptor pathways. AB - Peptides encoded by the urocortin II (Ucn II) gene were recently identified as new members of the corticotropin-releasing factor (CRF) family. Ucn II is a specific ligand for the type 2 CRF receptor. Using RT-PCR, DNA sequencing, and immunofluorescence staining, we report the expression of Ucn II mRNA in several human and mouse (m) neuronal cell lines. Using these neuronal cell lines, we provide evidence that exposure to glucocorticoid hormones increases mUcn II mRNA expression and promoter activation. The effect of glucocorticoids on mUcn II mRNA expression was tested in the Ucn II/glucocorticoid receptor-positive cell line NG108-15. The results demonstrate that mUcn II mRNA expression is up-regulated by dexamethasone in a dose- and time-dependent fashion. Computer analysis revealed the presence of 14 putative half-palindrome glucocorticoid response element sequences within 1.2 kb of the mUcn II 5' flanking region. Transfections with different fragments of the 5'-flanking region of the mUcn II gene fused to a luciferase reporter gene showed a promoter-dependent expression of the reporter gene and regulation by dexamethasone. Promoter deletion studies clarify the sufficient putative glucocorticoid response element site mediating this effect. The steroid hormone antagonist RU486 blocked the effect of dexamethasone on mUcn II mRNA expression and promoter activation, suggesting a direct glucocorticoid receptor-mediated effect of dexamethasone on mUcn II mRNA expression. Ucn II is expressed in vivo in the hypothalamus, brainstem, olfactory bulb, and pituitary. Low levels were also detected in the mouse cortex, hippocampus, and spinal cord. We demonstrated that mUcn II gene transcription was stimulated by glucocorticoid administration in vivo and inhibited by removal of glucocorticoids by adrenalectomy. Administration of dexamethasone to mice resulted in an increase of mUcn II levels in the hypothalamus and brainstem but not in the olfactory bulb region 12 h following ip injection. In light of our present data and the current literature, we propose a putative link between the CRF receptor 1 and CRF receptor 2 pathways. PMID- 12764079 TI - Functional brain imaging and human brain function. PMID- 12764080 TI - The underpinnings of the BOLD functional magnetic resonance imaging signal. PMID- 12764081 TI - Brain function and neurophysiological correlates of signals used in functional neuroimaging. PMID- 12764082 TI - Neuroimaging weighs in: humans meet macaques in "primate" visual cortex. PMID- 12764083 TI - Neuroimaging studies of attention: from modulation of sensory processing to top down control. PMID- 12764084 TI - Functional-anatomic correlates of control processes in memory. PMID- 12764085 TI - Visual cortex activity in early and late blind people. PMID- 12764086 TI - Functional correlations between neighboring neurons in the primate globus pallidus are weak or nonexistent. AB - The anatomical structure of the basal ganglia displays topographical organization and massive funneling of neuronal projections toward the globus pallidus as well as an axonal collateral system within this nucleus. This structure suggests the formation of correlations between the spiking activities of pallidal cells. Nevertheless, previous studies of remote neurons in the pallidum have reported uncorrelated spiking activity. These correlation results may be challenged, because remote pallidal neurons may be located in different pallidal territories. To further test the independence of pallidal activity, we studied the spiking activity of neighboring pairs recorded by the same electrodes. A narrow peak dominated the correlations of all pairs of neurons recorded on the same electrode. This type of peak is classically interpreted as a sign of strong common input. However, recent mathematical analysis shows that such peaks may derive from a technical inability to detect overlapping spikes by spike-sorting techniques. A long-term shallow trough in the correlation of neighboring neurons may also result from the same effect, which we have termed the "shadowing effect." A comparison of the expected shadowing effect with the actual correlations suggests that no real correlations exist between 93.9% of neighboring pallidal pairs. The remaining 6.1% of the pairs display symmetric long-term positive correlations centered on time 0. Thus, functional interactions between neighboring pallidal neurons do not display any significant differences from the interactions between physically remote neurons in this brain area. Moreover, the combination of anatomical data and current physiological results suggests an active decorrelating process performed in the basal ganglia. PMID- 12764087 TI - p38 mitogen-activated protein kinase is activated after a spinal nerve ligation in spinal cord microglia and dorsal root ganglion neurons and contributes to the generation of neuropathic pain. AB - The possible involvement of p38 mitogen-activated protein kinase activation in spinal cord and dorsal root ganglion (DRG) cells in the development of peripheral neuropathic pain has been explored. Ligation of the L5 spinal nerve (SNL) on one side in adult rats produces an early onset and long-lasting mechanical allodynia. This lesion results in activation of p38 in the L5 segment of the spinal cord, most prominently in the ipsilateral dorsal horn, starting soon after the lesion (<1 d) and persisting for >3 weeks. The activated p38 in the spinal cord is restricted entirely to microglia; phospho-p38 colocalizes only with the microglial marker OX-42 and not with either the neuronal marker neuronal-specific nuclear protein or the astrocyte marker GFAP. In contrast, SNL induces a delayed (>3 d) activation of p38 in the L5 DRG that occurs predominantly in neurons. Continuous injection of the p38 inhibitor 4-(4-fluorophenyl)-2-(4 methylsulfonylphenyl)-5-(4-pyridyl)-1H-imidazole (SB203580) via the intrathecal route, starting before the SNL surgery, reduces SNL-induced mechanical allodynia from day 1 to day 10, with maximal effects at early time points. Post-treatment with SB203580 starting on day 1 or on day 10 after surgery also reduces established mechanical allodynia. Because the reduction in neuropathic pain by p38 inhibition occurs before the appearance of p38 activation in DRG neurons, p38 activation in spinal cord microglia is likely to have a substantial role in the earliest phase of neuropathic pain. Coactivation of p38 in DRG neurons and spinal microglia may contribute to later phases of neuropathic pain. PMID- 12764088 TI - Apoptotic death of beta cells after optic nerve transection in adult cats. AB - We have revealed previously that the survival rate of beta cells of cat retinal ganglion cells (RGCs) rapidly decreased to 29% on day 7 after optic nerve transection, whereas that of alpha cells slowly decreased to 64% on day 14 (Watanabe et al., 2001). The reason that beta cells die more rapidly than alpha cells was not clear. In the present study, we tested the possibility that the rapid death of beta cells is attributable to apoptosis, as shown for some axotomized RGCs in rats. The following results were obtained. First, the proportion of pyknotic cells in Nissl-stained cat retinas started to increase sharply starting on day 4 and reached a peak on day 6 after optic nerve transection. The time course of occurrence of pyknotic cells corresponded well with that of the rapid death of axotomized beta cells. Secondly, the proportion of pyknotic cells was the highest in the area centralis (AC), in which beta cells are densely distributed. The preferential death of axotomized RGCs in the AC was also confirmed by terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end labeling staining in cross sections. Thirdly, after the intravitreal injection of caspase 3 inhibitor (z-DEVD-cmk) the survival of axotomized beta cells on day 7 was significantly enhanced, whereas no such survival-promoting effect was obtained in axotomized alpha cells. Taken together, these results suggest that the rapid death of axotomized beta cells is attributable mainly to apoptosis, which is mediated by caspase 3. PMID- 12764089 TI - Convergent mechanisms mediate preparatory states and repetition priming in the feeding network of Aplysia. AB - Improvement of behavioral responses to expected stimuli has been attributed to a change in the preparatory state. In this study, we take advantage of the accessibility of the nervous system of Aplysia and develop an in vitro analog of the preparatory state for feeding behaviors. We provide evidence that the change in the preparatory state may be elicited initially by activity of identified serotonergic metacerebral cells (MCCs). We demonstrate, however, that the preparatory state is maintained through MCC-independent repetition priming that is embedded in the properties of the behavior-generating network. Both MCC dependent and MCC-independent processes converge on the same site in the behavior generating network to mediate the change in the preparatory state. Thus, we propose a model of how multiple neuronal structures interact to elicit and maintain changes in the preparatory states in the CNS. PMID- 12764090 TI - Exploitation of astrocytes by glioma cells to facilitate invasiveness: a mechanism involving matrix metalloproteinase-2 and the urokinase-type plasminogen activator-plasmin cascade. AB - The presence of reactive astrocytes around glioma cells in the CNS suggests the possibility that these two cell types could be interacting. We addressed whether glioma cells use the astrocyte environment to modulate matrix metalloproteinase-2 (MMP-2), a proteolytic enzyme implicated in the invasiveness of glioma cells. We found that astrocytes in culture produce significant amounts of the pro-form of MMP-2 but undetectable levels of active MMP-2. However, after coculture with the U251N glioma line, astrocyte pro-MMP-2 was converted to the active form. The mechanism of pro-MMP-2 activation in glioma-astrocyte coculture was investigated and was found to involve the urokinase-type plasminogen activator (uPA)-plasmin cascade whereby uPA bound to uPA receptor (uPAR), leading to the conversion of plasminogen to plasmin. The latter cleaved pro-MMP-2 to generate its active form. Furthermore, key components (i.e., uPAR, uPA, and pro-MMP-2) were contributed principally by astrocytes, whereas the U251N glioma cells provided plasminogen. In correspondence with this biochemical cascade, the transmigration of U251N cells through Boyden invasion chambers coated with an extracellular matrix barrier was increased significantly in the presence of astrocytes, and this was inhibited by agents that disrupted the uPA-plasmin cascade. Finally, using resected human glioblastoma specimens, we found that tumor cells, but not astrocytes, expressed plasminogen in situ. We conclude that glioma cells exploit their astrocyte environment to activate MMP-2 and that this leads to the increased invasiveness of glioma cells. PMID- 12764091 TI - The alpha-latrotoxin mutant LTXN4C enhances spontaneous and evoked transmitter release in CA3 pyramidal neurons. AB - Alpha-latrotoxin (LTX) stimulates vesicular exocytosis by at least two mechanisms that include (1) receptor binding-stimulation and (2) membrane pore formation. Here, we use the toxin mutant LTX(N4C) to selectively study the receptor-mediated actions of LTX. LTX(N4C) binds to both LTX receptors (latrophilin and neurexin) and greatly enhances the frequency of spontaneous and miniature EPSCs recorded from CA3 pyramidal neurons in hippocampal slice cultures. The effect of LTX(N4C) is reversible and is not attenuated by La3+ that is known to block LTX pores. On the other hand, LTX(N4C) action, which requires extracellular Ca2+, is inhibited by thapsigargin, a drug depleting intracellular Ca2+ stores, by 2 aminoethoxydiphenyl borate, a blocker of inositol(1,4,5)-trisphosphate-induced Ca2+ release, and by U73122, a phospholipase C inhibitor. Furthermore, measurements using a fluorescent Ca2+ indicator directly demonstrate that LTX(N4C) increases presynaptic, but not dendritic, free Ca2+ concentration; this Ca2+ rise is blocked by thapsigargin, suggesting, together with electrophysiological data, that the receptor-mediated action of LTX(N4C) involves mobilization of Ca2+ from intracellular stores. Finally, in contrast to wild-type LTX, which inhibits evoked synaptic transmission probably attributable to pore formation, LTX(N4C) actually potentiates synaptic currents elicited by electrical stimulation of afferent fibers. We suggest that the mutant LTX(N4C), lacking the ionophore-like activity of wild-type LTX, activates a presynaptic receptor and stimulates Ca2+ release from intracellular stores, leading to the enhancement of synaptic vesicle exocytosis. PMID- 12764092 TI - Lighting up the senses: FM1-43 loading of sensory cells through nonselective ion channels. AB - We describe a novel mechanism for vital fluorescent dye entry into sensory cells and neurons: permeation through ion channels. In addition to the slow conventional uptake of styryl dyes by endocytosis, small styryl dyes such as FM1 43 rapidly and specifically label hair cells in the inner ear by entering through open mechanotransduction channels. This labeling can be blocked by pharmacological or mechanical closing of the channels. This phenomenon is not limited to hair cell transduction channels, because human embryonic kidney 293T cells expressing the vanilloid receptor (TRPV1) or a purinergic receptor (P2X2) rapidly take up FM1-43 when those receptor channels are opened and not when they are pharmacologically blocked. This channel permeation mechanism can also be used to label many sensory cell types in vivo. A single subcutaneous injection of FM1 43 (3 mg/kg body weight) in mice brightly labels hair cells, Merkel cells, muscle spindles, taste buds, enteric neurons, and primary sensory neurons within the cranial and dorsal root ganglia, persisting for several weeks. The pattern of labeling is specific; nonsensory cells and neurons remain unlabeled. The labeling of the sensory neurons requires dye entry through the sensory terminal, consistent with permeation through the sensory channels. This suggests that organic cationic dyes are able to pass through a number of different sensory channels. The bright and specific labeling with styryl dyes provides a novel way to study sensory cells and neurons in vivo and in vitro, and it offers new opportunities for visually assaying sensory channel function. PMID- 12764093 TI - Evidence for protein tyrosine phosphatase, tyrosine kinase, and G-protein regulation of the parallel fiber metabotropic slow EPSC of rat cerebellar Purkinje neurons. AB - The slow EPSP (sEPSP) or slow EPSC (sEPSC) at parallel fiber to Purkinje neuron synapses is attributable to a nonselective cation channel coupled to activation of metabotropic type 1 glutamate receptors (mGluR1s). Photorelease of L-glutamate in 1 msec from 4-methoxy-7-nitroindolinyl-or 7-nitroindolinyl-caged glutamate in cerebellar slices was used to isolate and study postsynaptic mechanisms coupling mGluR1 to the cation channel. L-Glutamate immediately activated a glutamate transporter current, followed by the slow mGluR1-activated conductance. Inhibitors of kinases, phosphatases, and G-proteins were tested on the peak glutamate-evoked currents. No effects of the inhibitors were seen on the initial glutamate transporter currents. In contrast, the later mGluR1 currents were either unaffected or enhanced by the protein tyrosine kinase (PTK) inhibitors PP1, K252a, and staurosporine were diminished or blocked by phosphatase inhibitors but were unaffected by inhibitors of serine-threonine kinases PKA, PKC, or PKG. The selective src-PTK inhibitor PP1 (10 microm intracellularly) potentiated submaximal mGluR1 currents evoked by low L-glutamate concentrations but had no effect on maximal responses (80 or 160 microm L-glutamate). L Glutamate-evoked mGluR1 currents and parallel fiber sEPSCs were reversibly and completely inhibited by protein tyrosine phosphatase (PTP) inhibitor bpV(phen) (50-200 microm) and by nonselective phosphatase inhibitor orthovanadate (0.5 or 1 mm). mGluR1 currents were completely inhibited by GDPbetaS applied intracellularly (5 mm). The results confirm a role for a GTPase postsynaptically, show that tyrosine phosphorylation inhibits mGluR1 coupling to the channel, and show that PTPs increase activation by tyrosine dephosphorylation most likely upstream of the sEPSP cation channel. PMID- 12764094 TI - Focal adhesion kinase is required, but not sufficient, for the induction of long term potentiation in dentate gyrus neurons in vivo. AB - Tyrosine kinase phosphorylation plays an important role in the induction of long term potentiation (LTP). Focal adhesion kinase (FAK) is a 125 kDa nonreceptor tyrosine kinase that shows decreased phosphorylation in fyn mutant mice, and Fyn plays a critical role in LTP induction. By examining the role of FAK involved in LTP induction in dentate gyrus in vivo with medial perforant path stimulation, we found that both FAK and mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) phosphorylation were increased significantly 5 and 10 min after LTP induction, whereas cAMP-responsive element binding protein (CREB) phosphorylation was increased 40 min later. Transfection of the dominant negative FAK mutant construct HA-FAK(Y397F) impaired LTP, whereas transfection of the constitutively activated form HA-FAK(Delta1-100) reduced the threshold for LTP induction. Transfection of HA-FAK(Delta1-100) by itself did not induce long lasting potentiation. Further, transfection of the HA-FAK(Y397F) construct decreased FAK, MAPK/ERK, and CREB phosphorylation, and the inhibition of MAPK/ERK decreased CREB phosphorylation. Moreover, blockade of NMDA receptor (NMDAR) did not decrease FAK, MAPK/ERK, and CREB phosphorylation although LTP induction was blunted by NMDAR antagonist. These biochemical changes were not associated with low-frequency stimulation either. Immunoprecipitation results revealed that tyrosine phosphorylation of NR2A and NR2B as well as the association of phosphorylated FAK with NR2A and NR2B was increased with LTP induction. These results together suggest that FAK is required, but not sufficient, for the induction of LTP in a NMDAR-independent manner and that MAPK/ERK and CREB are the downstream events of FAK activation. Further, FAK may interact with NR2A and NR2B to modulate LTP induction. PMID- 12764096 TI - Endocytosis and vesicle recycling at a ribbon synapse. AB - At ribbon synapses, where exocytosis is regulated by graded depolarization, vesicles can fuse very rapidly with the plasma membrane (complete discharge of the releasable pool in approximately 200 msec). Vesicles are also retrieved very rapidly (time constant of approximately 1 sec), leading us to wonder whether their retrieval uses an unusual mechanism. To study this, we exposed isolated bipolar neurons from goldfish retina to cationized ferritin. This electron-dense marker uniformly decorated the cell membrane and was carried into the cell during membrane retrieval. Endocytosis was activity-dependent and restricted to the synaptic terminal. The labeling pattern was consistent with direct retrieval from the plasma membrane of large, uncoated endosomes 60-200 nm in diameter. Even after extensive synaptic activity lasting several minutes, most of the ferritin remained in large endosomes and was present in only approximately 10% of the small vesicles that constitute the reserve pool. By contrast, after brief stimulation at a conventional terminal, ferritin did not reside in endosomes but was present in approximately 63% of the small vesicles. We suggest that the bipolar ribbon synapse sustains its rapid exocytosis by retrieving membrane in larger "bites" than the clathrin-dependent mechanism thought to dominate at conventional synapses. The resulting large endosomes bud off small vesicles, which reenter the reserve pool and finally the releasable pool. PMID- 12764095 TI - Inhibition of calpains prevents neuronal and behavioral deficits in an MPTP mouse model of Parkinson's disease. AB - The molecular mechanisms mediating degeneration of midbrain dopamine neurons in Parkinson's disease (PD) are poorly understood. Here, we provide evidence to support a role for the involvement of the calcium-dependent proteases, calpains, in the loss of dopamine neurons in a mouse model of PD. We show that administration of N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) evokes an increase in calpain-mediated proteolysis in nigral dopamine neurons in vivo. Inhibition of calpain proteolysis using either a calpain inhibitor (MDL-28170) or adenovirus-mediated overexpression of the endogenous calpain inhibitor protein, calpastatin, significantly attenuated MPTP-induced loss of nigral dopamine neurons. Commensurate with this neuroprotection, MPTP-induced locomotor deficits were abolished, and markers of striatal postsynaptic activity were normalized in calpain inhibitor-treated mice. However, behavioral improvements in MPTP-treated, calpain inhibited mice did not correlate with restored levels of striatal dopamine. These results suggest that protection against nigral neuron degeneration in PD may be sufficient to facilitate normalized locomotor activity without necessitating striatal reinnervation. Immunohistochemical analyses of postmortem midbrain tissues from human PD cases also displayed evidence of increased calpain-related proteolytic activity that was not evident in age matched control subjects. Taken together, our findings provide a potentially novel correlation between calpain proteolytic activity in an MPTP model of PD and the etiology of neuronal loss in PD in humans. PMID- 12764097 TI - NMDA receptor overactivation inhibits phospholipid synthesis by decreasing choline-ethanolamine phosphotransferase activity. AB - Overactivation of NMDA receptors is believed to induce neuronal death by increasing phospholipid hydrolysis and subsequent degradation. We showed previously that NMDA releases choline and inhibits incorporation of [3H]choline into phosphatidylcholine before excitotoxic neuronal death. On the basis of these results, we hypothesized that excitotoxicity results from inhibition of synthesis rather than from increased degradation of phospholipids. We now investigated the effect of NMDA receptor overactivation on synthesis and degradation of major membrane phospholipids in the early stages of the excitotoxic process. Exposure of cortical neurons to neurotoxic concentrations of NMDA increased extracellular choline and activated hydrolysis of phosphatidylcholine and phosphatidylinositol by phospholipase A2 but did not induce significant degradation of phosphatidylcholine, phosphatidylinositol, phosphatidylethanolamine, or phosphatidylserine. In contrast, NMDA strongly reduced the incorporation of [3H]choline and [3H]ethanolamine into their respective phospholipids. Metabolic labeling experiments in whole cells showed that NMDA receptor overactivation does not modify the activity of phosphocholine or phosphoethanolamine cytidylyltransferases but strongly inhibits choline-ethanolamine phosphotransferase activity. This effect was observed well before any significant membrane damage and cell death. Moreover, cholinephosphotransferase activity was lower in microsomes from NMDA-treated cells. These results show that membrane damage by NMDA is preceded by inhibition of phospholipid synthesis and not by phospholipid degradation in the early stages of the excitotoxic process, and that NMDA receptor overactivation decreases phosphatidylcholine and phosphatidylethanolamine synthesis by inhibiting choline ethanolaminophosphotransferase activity. PMID- 12764098 TI - In vivo whole-cell recording of odor-evoked synaptic transmission in the rat olfactory bulb. AB - One of the first steps in the coding of olfactory information is the transformation of synaptic input to action potential firing in mitral and tufted (M/T) cells of the mammalian olfactory bulb. However, little is known regarding the synaptic mechanisms underlying this process in vivo. In this study, we examined odor-evoked response patterns of M/T and granule cells using whole-cell recording in anesthetized, freely breathing rats. We find that odor-evoked excitatory responses in M/T cells typically consist of bursts of action potentials coupled to the approximately 2 Hz respiration rhythm. Odor-evoked, rhythmic M/T cell excitation is reliable during odor presentation (2-4 sec); in contrast, both excitatory responses of granule cells and M/T cell lateral inhibition adapt quickly after the first respiration cycle in the presence of odorants. We also find that the amplitude and initial slope of odor-evoked synaptic excitation play an important role in regulating the timing of M/T cell spikes. Furthermore, differences in odor concentration alter the shape of odor evoked excitatory synaptic responses, the latency of M/T cell spikes, and the timing of M/T cell lateral inhibition. PMID- 12764099 TI - Differential activation of extracellular signal-regulated protein kinase in primary afferent neurons regulates brain-derived neurotrophic factor expression after peripheral inflammation and nerve injury. AB - To investigate the intracellular signal transduction pathways involved in regulating the gene expression of brain-derived neurotrophic factor (BDNF) in primary afferent neurons, we examined the activation of extracellular signal regulated protein kinase (ERK) in dorsal root ganglion (DRG) neurons after peripheral inflammation and sciatic nerve transection. Peripheral inflammation induced an increase in the phosphorylation of ERK, mainly in tyrosine kinase A containing small-to-medium-diameter DRG neurons. The treatment of the mitogen activated protein kinase (MAPK) kinase 1/2 inhibitor U0126 reversed the pain hypersensitivity and the increase in phosphorylated-ERK (p-ERK) and BDNF in DRG neurons induced by complete Freund's adjuvant. On the other hand, axotomy induced the activation of ERK mainly in medium-and large-sized DRG neurons and in satellite glial cells. U0126 suppressed the axotomy-induced autotomy behavior and reversed the increase in p-ERK and BDNF. The intrathecal application of nerve growth factor (NGF) induced an increase in the number of p-ERK-and BDNF-labeled cells, mainly small neurons, and the application of anti-NGF induced an increase in p-ERK and BDNF in some medium-to-large-diameter DRG neurons. The activation of MAPK in the primary afferents may occur in different populations of DRG neurons after peripheral inflammation and axotomy, respectively, through alterations in the target-derived NGF. These changes, including the changes in BDNF expression, might be involved in the pathophysiological changes in primary afferent neurons. PMID- 12764100 TI - Neuroprotection by the endogenous cannabinoid anandamide and arvanil against in vivo excitotoxicity in the rat: role of vanilloid receptors and lipoxygenases. AB - Type 1 vanilloid receptors (VR1) have been identified recently in the brain, in which they serve as yet primarily undetermined purposes. The endocannabinoid anandamide (AEA) and some of its oxidative metabolites are ligands for VR1, and AEA has been shown to afford protection against ouabain-induced in vivo excitotoxicity, in a manner that is only in part dependent on the type 1 cannabinoid (CB1) receptor. In the present study, we assessed whether VR1 is involved in neuroprotection by AEA and by arvanil, a hydrolysis-stable AEA analog that is a ligand for both VR1 and CB1. Furthermore, we assessed the putative involvement of lipoxygenase metabolites of AEA in conveying neuroprotection. Using HPLC and gas chromatography/mass spectroscopy, we demonstrated that rat brain and blood cells converted AEA into 12-hydroxy-N-arachidoylethanolamine (12 HAEA) and 15-hydroxy-N-arachidonoylethanolamine (15-HAEA) and that this conversion was blocked by addition of the lipoxygenase inhibitor nordihydroguaiaretic acid. Using magnetic resonance imaging we show the following: (1) pretreatment with the reduced 12-lipoxygenase metabolite of AEA, 12-HAEA, attenuated cytotoxic edema formation in a CB1 receptor-independent manner in the acute phase after intracranial injection of the Na+/K+-ATPase inhibitor ouabain; (2) the reduced 15-lipoxygenase metabolite, 15-HAEA, enhanced the neuroprotective effect of AEA in the acute phase; (3) modulation of VR1, as tested using arvanil, the VR1 agonist capsaicin, and the antagonist capsazepine, leads to neuroprotective effects in this model, and arvanil is a potent neuroprotectant, acting at both CB1 and VR1; and (4) the in vivo neuroprotective effects of AEA are mediated by CB1 but not by lipoxygenase metabolites or VR1. PMID- 12764101 TI - Expression and function of chloride transporters during development of inhibitory neurotransmission in the auditory brainstem. AB - Glycine and GABA, the dominant inhibitory neurotransmitters in the CNS, assume a depolarizing role in early development, leading to increased cytoplasmic Ca2+ levels and action potentials. The effect is thought to be of some significance for maturation. The depolarization is caused by Cl- efflux, and chloride transporters contribute to the phenomenon by raising the intracellular Cl- concentration ([Cl-]i) above equilibrium, thereby generating an outward-directed electrochemical gradient for Cl-. In mature neurons, the [Cl-]i is reduced below equilibrium, thus rendering glycine activity hyperpolarizing. Here, we investigated the temporal expression of the K-Cl cotransporter KCC2 and the Na-K Cl cotransporter NKCC1 in the lateral superior olive (LSO) of rats and mice. The two cation cotransporters normally extrude and accumulate Cl-, respectively. As evidenced by several methods, KCC2 mRNA was present in LSO neurons during both the depolarizing and hyperpolarizing periods. Western blots confirmed a constant level of KCC2 in the brainstem, and immunohistochemistry showed that the protein is diffusely distributed within neonatal LSO neurons, becoming integrated into the plasma membrane only with increasing age. The glycine reversal potential in KCC2 knock-out mice differed significantly from that determined in wild-type controls at postnatal day 12 (P12) but not at P3, demonstrating that KCC2 is not active in neonates, despite its early presence. NKCC1 mRNA was not detected during the depolarizing phase in the LSO, implying that this transporter does not contribute to the high [Cl-]i. Our results reveal major differences in the development of [Cl-]i regulation mechanisms seen in brainstem versus forebrain regions. PMID- 12764102 TI - Synapse number and synaptic efficacy are regulated by presynaptic cAMP and protein kinase A. AB - The mechanisms by which neurons regulate the number and strength of synapses during development and synaptic plasticity have not yet been defined fully. This lack of fundamental knowledge in the fields of neurodevelopment and synaptic plasticity can be attributed, in part, to compensatory mechanisms by which neurons accommodate for the loss of function in their synaptic partners. This is generally achieved either by scaling up neuronal transmitter release capabilities or by enhancing the postsynaptic responsiveness. Here, we demonstrate that regulation of synaptic strength and number between identified Lymnaea neurons visceral dorsal 4 (VD4, the presynaptic cell) and left pedal dorsal 1 (LPeD1, the postsynaptic cell) requires presynaptic activation of a cAMP-PKA-dependent signal. Experimental activation of the cAMP-PKA pathway resulted in reduced synaptic efficacy, whereas inhibition of the cAMP-PKA cascade permitted hyperinnervation and an overall enhancement of synaptic strength. Because synaptic transmission between VD4 and LPeD1 does not require a cAMP-PKA pathway, our data show that these messengers may play a novel role in regulating the synaptic efficacy during early synaptogenesis and plasticity. PMID- 12764103 TI - Frequency modulation of synchronized Ca2+ spikes in cultured hippocampal networks through G-protein-coupled receptors. AB - Synchronized spontaneous Ca2+ spikes in networked neurons represent periodic burst firing of action potentials, which are believed to play a major role in the development and plasticity of neuronal circuitry. How these network activities are shaped and modulated by extrinsic factors during development, however, remains to be studied. Here we report that synchronized Ca2+ spikes among cultured hippocampal neurons can be modulated by two small factors that act on G protein-coupled receptors (GPCRs): the neuropeptide PACAP (pituitary adenylate cyclase-activating polypeptide) and the chemokine SDF-1 (stromal cell-derived factor-1). PACAP effectively increases the frequency of the synchronized Ca2+ spikes when applied acutely; the PACAP potentiation of Ca2+ spikes requires the activation of the PACAP-specific PAC1 GPCRs and is mediated by the activation of cAMP signaling pathway. SDF-1, on the other hand, significantly reduces the frequency of these Ca2+ spikes through the activation of its specific GPCR CXCR4; the inhibitory action of SDF-1 is mediated by the inhibition of cAMP pathway through the Gi component of GPCRs. Taken together, these results demonstrate that synchronized neuronal network activity can be effectively modulated by physiologically and developmentally relevant small factors that act on GPCRs to target the cAMP pathway. Such modulation of neuronal activity through GPCRs may represent a significant mechanism that underlies the neuronal plasticity during neural development and functioning. PMID- 12764105 TI - Enhancement of associative long-term potentiation by activation of beta adrenergic receptors at CA1 synapses in rat hippocampal slices. AB - The objective of this study was to evaluate the role of beta-adrenergic receptors in modulating associative long-term potentiation (LTP) induced at CA1 synapses. Two independent Schaffer collateral pathways were stimulated in hippocampal slices. The field EPSP (fEPSP) response evoked in one pathway (the weak pathway) was small, whereas a large response, usually 80-90% of the maximum, was evoked in the strong pathway. After recording of the baseline fEPSP evoked at 0.033 Hz, LTP of the weak pathway could be associatively induced by paired stimulation of the weak and strong pathways 100 times at 6 sec intervals, with stimulation of the weak pathway preceded 3-10 msec. However, pairing protocols with an interval between stimulation of the two pathways >10 msec resulted in no LTP. The induced LTP was NMDA receptor dependent, because 50 microm D,L-APV blocked its induction. Bath application of 1 microm isoproterenol enhanced LTP by increasing the window of the stimulation interval up to 15 msec but did not affect the magnitude of the LTP induced by pairing protocols with intervals <10 msec. Similar results were obtained when the experiments were repeated using whole-cell recording. These results suggest that activation of beta-adrenergic receptors can enhance associative LTP by increasing the width of the time window rather than the magnitude of the LTP. Enhancement of LTP by beta-adrenergic receptors was blocked in slices by pretreatment with inhibitors of protein kinase A or mitogen activated protein kinase, suggesting that these signaling cascades are involved in this process. PMID- 12764104 TI - Increased expression of brain-derived neurotrophic factor preserves retinal function and slows cell death from rhodopsin mutation or oxidative damage. AB - There are no effective treatments for inherited retinal degenerations, which are prevalent causes of visual disability. Several proteins promote the survival of various types of neurons, and increasing expression of one or more of these survival factors is a promising strategy for a new treatment. Studies examining the effects of intravitreous injections of brain-derived neurotrophic factor (BDNF) in models of inherited retinal degenerations have suggested that BDNF has little survival-promoting activity for photoreceptors. In this study, we generated double transgenic mice with doxycycline-inducible expression of BDNF in the retina. In a model of primary rod photoreceptor degeneration, expression of BDNF resulted in significant delay in photoreceptor cell death and maintenance of retinal function assessed by electroretinogram recordings. Expression of BDNF also caused strong protection of photoreceptors from oxidative damage-induced cell death. These data suggest that continuous expression of BDNF, unlike intravitreous injections, results in morphologic and functional benefit in animal models of inherited retinal degeneration. Double transgenic mice with inducible expression of survival factors provide valuable tools for selection of survival factor candidates for gene therapy. PMID- 12764106 TI - Serotonin 1A receptor agonists reverse respiratory abnormalities in spinal cord injured rats. AB - Contusion spinal cord injury (SCI) at T8 produces respiratory abnormalities in conscious rats breathing room air and challenged with CO2. In seeking ways to improve respiration after SCI, we tested drugs that stimulate serotonin 1A (5 HT1A) receptors, based on our previous findings that these agents can counteract respiratory depression produced by morphine overdose. Respiratory function was measured with a head-out plethysmograph system in conscious rats. T8 SCI rats (n = 5) showed decreased tidal volume (Vt; 0.90 +/- 0.02-0.66 +/- 0.03 ml; p < 0.05) and increased respiratory rate (f;91 +/- 3.7-132 +/- 5.7 breaths/min; p < 0.05) with room air ventilation at 24 hr after injury. They also exhibited a diminished response to the respiratory stimulating effect of 7% CO2; minute ventilation increased to 250 +/- 17 ml/min before, but only to 162 +/- 15 ml/min at 24 hr after SCI (p < 0.05). Respiratory deficits during room air ventilation were also observed at 7 d after injury (n = 3). Treatment with the 5-HT1A receptor agonist 8-hydroxy-2-(di-n-propylmino)tetralin (8-OH-DPAT; 250 microg/kg, i.p.) at 24 hr (n = 5) or 7 d (n = 3) after injury normalized Vt, f, and the respiratory response to 7% CO2. Identical results were obtained with another 5-HT1A receptor agonist, buspirone (1.5 mg/kg, i.p.; n = 3). In contrast, intraperitoneal saline vehicle administration (n = 5) showed no beneficial effects on SCI-impaired respiration. Finally, pretreatment with a specific antagonist of 5-HT1A receptors, 4-iodo-N-[2-[4-(methoxyphenyl)-1-piperazinyl]ethyl]-N-2-pyridinyl benzamide (3 mg/kg, i.p.; n = 3) given 20 min before 8-OH-DPAT, prevented 8-OH DPAT from restoring respiration to normal. Our results demonstrate that drugs that stimulate 5-HT1A receptors counteract respiratory abnormalities in conscious rats after SCI. PMID- 12764107 TI - Transmembrane sema4E guides branchiomotor axons to their targets in zebrafish. AB - Class 4 semaphorins are a large class of transmembrane proteins that contain a sema domain and that are expressed in the CNS, but their in vivo neural function is unknown. In zebrafish, the epithelial cells that line the pharyngeal arches express Sema4E. Extension of branchiomotor axons along the mesenchymal cells bounded by these epithelial cells suggests that Sema4E may act as a repulsive guidance molecule to restrict the branchiomotor axons to the mesenchymal cells. To test this hypothesis, Sema4E was misexpressed in hsp70 promoter-regulated transgenic zebrafish in which sema4E was heat-inducible, and Sema4E was knocked down by injection of antisense morpholino oligonucleotides that acted specifically against Sema4E. Ubiquitous induction of Sema4E retarded outgrowth by the facial and gill branchiomotor axons significantly. Furthermore, outgrowth by gill motor axons was specifically inhibited when Sema4E-expressing transgenic cells were transplanted to their pathway in nontransgenic host embryos. Morpholino knockdown of Sema4E caused facial motor axons to defasciculate and follow aberrant pathways. These results show that Sema4E is repulsive for facial and gill motor axons and functions as a barrier for these axons within the pharyngeal arches. PMID- 12764108 TI - Location and size of dopaminergic and serotonergic cell populations are controlled by the position of the midbrain-hindbrain organizer. AB - Midbrain dopaminergic and hindbrain serotonergic neurons play an important role in the modulation of behavior and are involved in a series of neuropsychiatric disorders. Despite the importance of these cells, little is known about the molecular mechanisms governing their development. During embryogenesis, midbrain dopaminergic neurons are specified rostral to the midbrain-hindbrain organizer (MHO), and hindbrain serotonergic neurons are specified caudal to it. We report that in transgenic mice in which Otx2 and accordingly the MHO are shifted caudally, the midbrain dopaminergic neuronal population expands to the ectopically positioned MHO and is enlarged. Complementary, the extension of the hindbrain serotonergic cell group is decreased. These changes are preserved in adulthood, and the additional, ectopic dopaminergic neurons project to the striatum, which is a proper dopaminergic target area. In addition, in mutants in which Otx2 and the MHO are shifted rostrally, dopaminergic and serotonergic neurons are relocated at the newly positioned MHO. However, in these mice, the size ratio between these two cell populations is changed in favor of the serotonergic cell population. To investigate whether the position of the MHO during embryogenesis is also of functional relevance for adult behavior, we tested mice with a caudally shifted MHO and report that these mutants show a higher locomotor activity. Together, we provide evidence that the position of the MHO determines the location and size of midbrain dopaminergic and hindbrain serotonergic cell populations in vivo. In addition, our data suggest that the position of the MHO during embryogenesis can modulate adult locomotor activity. PMID- 12764109 TI - The growth arrest-specific gene product Gas6 promotes the survival of human oligodendrocytes via a phosphatidylinositol 3-kinase-dependent pathway. AB - Microarray analysis revealed that transcripts for the Axl and Mer receptor tyrosine kinases are expressed at high levels in O4+-immunopanned oligodendrocytes isolated from second trimester human fetal spinal cord. In humans the sole known ligand for the Axl/Rse/Mer kinases is growth arrest specific gene 6 (Gas6), which in the CNS is secreted by neurons and endothelial cells. We hypothesized that Gas6 is a survival factor for oligodendrocytes and receptor activation signals downstream to the phosphatidylinositol 3 (PI3) kinase/Akt pathway to increase cell survival in the absence of cell proliferation. To test this hypothesis, we grew enriched human oligodendrocytes for 6 d on a monolayer of NIH3T3 cells stably expressing Gas6. CNP+ oligodendrocytes on Gas6-secreting 3T3 cells had more primary processes and arborizations than those plated solely on 3T3 cells. Also, a twofold increase in CNP+ and MBP+ oligodendrocytes was observed when they were plated on the Gas6 secreting cells. The effect was abolished in the presence of Axl-Fc but remained unchanged in the presence of the irrelevant receptor fusion molecule TrkA-Fc. A significant decrease in CNP+/TUNEL+ oligodendrocytes was observed when recombinant human Gas6 (rhGas6) was administered to oligodendrocytes plated on poly-L-lysine, supporting a role for Gas6 signaling in oligodendrocyte survival during a period of active myelination in human fetal spinal cord development. PI3 kinase inhibitors blocked the anti-apoptotic effect of rhGas6, whereas a MEK/ERK inhibitor had no effect. Thus Gas6 sustains human fetal oligodendrocyte viability by receptor activation and downstream signaling via the PI3-kinase/Akt pathway. PMID- 12764110 TI - Delayed systemic Nogo-66 receptor antagonist promotes recovery from spinal cord injury. AB - Traumatized axons possess an extremely limited ability to regenerate within the adult mammalian CNS. The myelin-derived axon outgrowth inhibitors Nogo, oligodendrocyte-myelin glycoprotein, and myelin-associated glycoprotein, all bind to an axonal Nogo-66 receptor (NgR) and at least partially account for this lack of CNS repair. Although the intrathecal application of an NgR competitive antagonist at the time of spinal cord hemisection induces significant regeneration of corticospinal axons, such immediate local therapy may not be as clinically feasible for cases of spinal cord injury. Here, we consider whether this approach can be adapted to systemic therapy in a postinjury therapeutic time window. Subcutaneous treatment with the NgR antagonist peptide NEP1-40 (Nogo extracellular peptide, residues 1-40) results in extensive growth of corticospinal axons, sprouting of serotonergic fibers, upregulation of axonal growth protein SPRR1A (small proline-rich repeat protein 1A), and synapse re formation. Locomotor recovery after thoracic spinal cord injury is enhanced. Furthermore, delaying the initiation of systemic NEP1-40 administration for up to 1 week after cord lesions does not limit the degree of axon sprouting and functional recovery. This indicates that the regenerative capacity of transected corticospinal tract axons persists for weeks after injury. Systemic Nogo-66 receptor antagonists have therapeutic potential for subacute CNS axonal injuries such as spinal cord trauma. PMID- 12764111 TI - Vasopressin-induced cytoplasmic and nuclear calcium signaling in embryonic cortical astrocytes: dynamics of calcium and calcium-dependent kinase translocation. AB - The present study sought to determine the downstream consequences of V1a vasopressin receptor (V1aR) activation of Ca2+ signaling in cortical astrocytes. Results of these analyses demonstrated that V1aR activation led to a marked increase in both cytoplasmic and nuclear Ca2+. We also investigated V1aR activation of Ca2+-activated signaling kinases, protein kinase C (PKC), Ca2+/calmodulin-dependent protein kinase II (CaMKII), and the mitogen-activated protein (MAP) kinases [MAPK and extracellular signal-regulated kinases 1 and 2 (ERK1/2)], their localization within cytoplasmic and nuclear compartments, and activation of their downstream nuclear target, the transcription factor cAMP response element-binding protein (CREB). Results of these analyses demonstrated that V1aR activation led to a significant rise in PKC, CaMKII, and ERK1/2 activation, with CaMKII and ERK1/2 demonstrating dynamic transport between cytoplasmic and nuclear compartments. Although no evidence of PKC translocation was apparent, PKC and CaMKs were required for activation and nuclear translocation of ERK1/2. Subsequent to CaMKII and ERK1/2 translocation to the nucleus, CREB activation occurred and was found to be dependent on upstream activation of ERK1/2 and CaMKs. These data provide the first systematic analysis of the V1aR-induced Ca2+ signaling cascade in cortical astrocytes. In addition, results of this study introduce a heretofore unknown effect of vasopressin, dynamic Ca2+ signaling between the cytoplasm and nucleus that leads to comparable dynamics of kinase activation and shuttling between cytoplasmic and nuclear compartments. Implications for development and regeneration induced by V1aR activation of CREB-regulated gene expression in cortical astrocytes are discussed. PMID- 12764112 TI - Multiple cell populations in the early postnatal subventricular zone take distinct migratory pathways: a dynamic study of glial and neuronal progenitor migration. AB - Neural progenitors in the subventricular zone (SVZ) of the postnatal rat forebrain give rise to either olfactory interneurons or glia. To investigate the overall patterns of progenitor movement, we labeled neonatal rat SVZ cells by stereotactic injection of a GFP-encoding retrovirus into the SVZ at various coronal levels. We then studied the movements of labeled cells by time-lapse videomicroscopy in living brain slices cut in different orientations. We observed two migration patterns: (1) progenitors migrated radially into the overlying white matter and cortex, but only at the level of viral injection; these were previously shown to give rise to astrocytes and oligodendrocytes, (2) progenitors migrated in a bidirectional, rostrocaudal pattern along the entire extent of the SVZ; many of these cells eventually migrated into the olfactory bulb and developed into interneurons, but they did not turn to migrate radially out of the SVZ until they reached the olfactory bulb. Video imaging showed apparent boundaries to migration between the SVZ and adjacent structures. These observations indicate that there are at least two distinct migratory pathways within the SVZ used differentially by immature neurons and glia. PMID- 12764113 TI - A functional gamma-band defined by stimulus-dependent synchronization in area 18 of awake behaving cats. AB - During the past decade, numerous studies have demonstrated stimulus-specific synchronization of neuronal activity in the gamma-frequency range. However, it appears that the different analyses are based on widely different assumptions about which frequency range to investigate. Therefore, the term "gamma synchronization" refers to an inhomogeneous spectrum of definitions and corresponding frequency bands. Moreover, most studies have been performed in anesthetized animals or in awake animals by use of fixation paradigms. Thus, it is difficult to relate these results to alert animals behaving under natural conditions. Here, we investigate stimulus specific synchronization in primary visual cortex of awake cats in a tracking paradigm. We record local field potentials and multiunit activity simultaneously from multiple electrodes. (1) We demonstrate that visual stimulation induces neuronal synchronization in a broad frequency range reaching well above 100 Hz. (2) We derive a functional gamma-band based on an objective criterion: We show that synchronization of neuronal activity is optimally orientation-tuned when a broad frequency band is considered. This band starts above 40 Hz, a frequency that is typically related to the term gamma-synchronization, and extends to very high frequencies. Interestingly, the frequency of maximum synchronization is different from the frequency at which synchronization is most stimulus specific. (3) We demonstrate synchronization of neuronal activity in a distinct low-frequency band with different properties suggesting separate functional roles of low- and high frequency synchronization. PMID- 12764114 TI - Osmotic regulation of estrogen receptor-beta in rat vasopressin and oxytocin neurons. AB - The vasopressin (VP) magnocellular neurosecretory cells (MNCs) in the supraoptic and paraventricular (PVN) nuclei are regulated by estrogen and exhibit robust expression of estrogen receptor (ER)-beta. In contrast, only approximately 7.5% of oxytocin (OT) MNCs express ER-beta. We examined the osmotic regulation of ER beta mRNA expression in MNCs using quantitative in situ hybridization histochemistry. Hyper-osmolality induced via 2% hypertonic saline ingestion significantly decreased, whereas sustained hypo-osmolality induced via d-d arginine VP and liquid diet increased ER-beta mRNA expression in MNCs (p < 0.05). Thus, the expression of ER-beta mRNA correlated inversely with changes in plasma osmolality. Because hyper-osmolality is a potent stimulus for VP and OT release, this suggests an inhibitory role for ER-beta in MNCs. Immunocytochemistry demonstrated that the decrease in ER-beta mRNA was translated into depletion of receptor protein content in hyper-osmotic animals. Numerous MNCs were positive for ER-beta in control animals, but they were virtually devoid of ER-beta immunoreactivity (IR) in hyper-osmotic animals. The osmotically induced decrease in ER-beta expression was selective for MNCs because ER-beta-IR remained unaltered in PVN parvocellular neurons. Plasma estradiol and testosterone were not correlated with ER-beta mRNA expression after osmotic manipulation, suggesting that ER-beta expression was not driven by ligand availability. Expression of FOS-IR in MNCs with attenuated ER-beta-IR, and the absence of FOS IR in parvocellular neurons that retain ER-beta-IR suggest a role for neuronal activation in the regulation of ER-beta expression in MNCs. Thus, osmotic modulation of ER-beta expression in MNCs may augment or attenuate an inhibitory effect of gonadal steroids on VP release. PMID- 12764115 TI - Vasopressin differentially modulates non-NMDA receptors in vasopressin and oxytocin neurons in the supraoptic nucleus. AB - Magnocellular neurons of the supraoptic nucleus release the neuropeptides oxytocin and vasopressin from their dendrites to regulate their synaptic inputs. This study aims to determine the cellular mechanism by which vasopressin modulates excitatory synaptic transmission. Presumably by electroporation through perforated patch, we were able to successfully introduce biocytin into cells in which we performed an electrophysiological study. This method enabled us to determine that roughly half of the recorded neurons were immunoreactive to oxytocin-associated neurophysin and showed two characteristic features: an inward rectification and a sustained outward rectification. The remaining half showed a linear voltage-current relationship and was immunoreactive to vasopressin associated neurophysin. Using these electrophysiological characteristics and post hoc immunohistochemistry to identify vasopressin or oxytocin neurons, we found that vasopressin decreased evoked EPSCs in vasopressin neurons while increasing EPSCs in oxytocin neurons. In both types of neurons, EPSC decay constants were not affected, indicating that desensitization of non-NMDA receptors did not underlie the EPSC amplitude change. In vasopressin neurons, both vasopressin and a V1a receptor agonist, F-180, decreased AMPA-induced currents, an effect blocked by a V1a receptor antagonist SR49059. In oxytocin neurons, AMPA-induced currents were facilitated by vasopressin, whereas F-180 had no effect. An oxytocin receptor antagonist blocked the facilitatory effect of vasopressin. Thus, we conclude that vasopressin inhibits EPSCs in vasopressin neurons via postsynaptic V1a receptors, whereas it facilitates EPSCs in oxytocin neurons through oxytocin receptors. PMID- 12764117 TI - Rapid motor learning in the translational vestibulo-ocular reflex. AB - Motor learning was induced in the translational vestibulo-ocular reflex (TVOR) when monkeys were repeatedly subjected to a brief (0.5 sec) head translation while they tried to maintain binocular fixation on a visual target for juice rewards. If the target was world-fixed, the initial eye speed of the TVOR gradually increased; if the target was head-fixed, the initial eye speed of the TVOR gradually decreased. The rate of learning acquisition was very rapid, with a time constant of approximately 100 trials, which was equivalent to <1 min of accumulated stimulation. These learned changes were consolidated over >or=1 d without any reinforcement, indicating induction of long-term synaptic plasticity. Although the learning generalized to targets with different viewing distances and to head translations with different accelerations, it was highly specific for the particular combination of head motion and evoked eye movement associated with the training. For example, it was specific to the modality of the stimulus (translation vs rotation) and the direction of the evoked eye movement in the training. Furthermore, when one eye was aligned with the heading direction so that it remained motionless during training, learning was not expressed in this eye, but only in the other nonaligned eye. These specificities show that the learning sites are neither in the sensory nor the motor limb of the reflex but in the sensory-motor transformation stage of the reflex. The dependence of the learning on both head motion and evoked eye movement suggests that Hebbian learning may be one of the underlying cellular mechanisms. PMID- 12764118 TI - Nitric oxide-mediated cortical activation: a diffuse wake-up system. AB - Nitric oxide (NO) has been implicated in some of the central pathways engaged in the regulation of the sleep-wake cycle. The existence of nitric oxide synthase (NOS) in the cholinergic basal forebrain (BF) cells projecting to the cortex suggests a role for NO in the activation induced by the BF during arousal. We tested, in the anesthetized cat, the hypothesis that inhibition of NOS would decrease the ability of BF cholinergic fibers to induce cortical activation. In control conditions, BF stimulation evoked an awake-like EEG pattern (i.e., a decrease in the low-frequency-high-amplitude oscillatory activity and an increase in the high-frequency-low-amplitude activity). After blocking NOS activity, the capacity of BF stimulation to induce cortical activation was strongly impaired. Furthermore, voltammetric measurements of NO levels revealed an increase in cortical NO after BF stimulation, also blocked by systemic NOS inhibition. These results indicate that the blockade of NOS activity significantly reduces the ability of BF stimulation to induce changes in the EEG pattern and suggest a role for NO in the BF-cholinergic system implicated in arousal mechanisms. PMID- 12764116 TI - A1 receptor and adenosinergic homeostatic regulation of sleep-wakefulness: effects of antisense to the A1 receptor in the cholinergic basal forebrain. AB - We hypothesized that adenosine, acting via the A1 receptor, is a key factor in the homeostatic control of sleep. The increase in extracellular levels of adenosine during prolonged wakefulness is thought to facilitate the transition to sleep by reducing the discharge activity of wakefulness-promoting neurons in the basal forebrain. Adenosine A1 receptor control of the homeostatic regulation of sleep was tested by microdialysis perfusion of antisense oligonucleotides against the mRNA of the A1 receptor in the magnocellular cholinergic region of the basal forebrain of freely behaving rats. After microdialysis perfusion of A1 receptor antisense in the basal forebrain, spontaneous levels of sleep-wakefulness showed a significant reduction in non-rapid eye movement (REM) sleep with an increase in wakefulness. After 6 hr of sleep deprivation, the antisense-treated animals spent a significantly reduced amount of time in non-REM sleep, with postdeprivation recovery sleep hours 2-5 showing a reduction of approximately 50-60%. There was an even greater postdeprivation reduction in delta power (60-75%) and a concomitant increase in wakefulness. All behavioral state changes returned to control (baseline) values after the cessation of antisense administration. Control experiments with microdialysis perfusion of nonsense (randomized antisense) oligonucleotides and with artificial CSF showed no effect during postdeprivation recovery sleep or spontaneously occurring behavioral states. Antisense to the A1 receptor suppressed A1 receptor immunoreactivity but did not show any neurotoxicity as visualized by Fluoro-Jade staining. These data support our hypothesis that adenosine, acting via the A1 receptor, in the basal forebrain is a key component in the homeostatic regulation of sleep. PMID- 12764119 TI - Error monitoring using external feedback: specific roles of the habenular complex, the reward system, and the cingulate motor area revealed by functional magnetic resonance imaging. AB - The dopaminergic system has been shown to be involved in the processing of rewarding stimuli, specifically of errors in reward prediction, in animal studies as well as in recent neuroimaging studies in humans. Furthermore, a specific role of dopamine in the human homolog of the rostral cingulate motor area (rCMA) was proposed in a recent model of error detection. Negative feedback as well as self detected errors elicit a negative event-related brain potential probably generated in the rCMA. We performed two experiments using functional magnetic resonance imaging to investigate the brain activity related to negative and positive feedback in a dynamically adaptive motion prediction task. Whereas positive feedback raised hemodynamic activity in the ventral striatum (nucleus accumbens), negative feedback activated the rCMA, the inferior anterior insula, and the epithalamus (habenular complex). These data demonstrate the role of the habenular complex in the control of the human reward system, a function previously hypothesized on the basis of animal research. The rCMA reacted only to errors with negative feedback but not to errors without feedback, which ruled out an influence of response conflict or uncertainty on its role in error detection by external signals. PMID- 12764120 TI - Conscious expectation and unconscious conditioning in analgesic, motor, and hormonal placebo/nocebo responses. AB - The placebo and nocebo effect is believed to be mediated by both cognitive and conditioning mechanisms, although little is known about their role in different circumstances. In this study, we first analyzed the effects of opposing verbal suggestions on experimental ischemic arm pain in healthy volunteers and on motor performance in Parkinsonian patients and found that verbally induced expectations of analgesia/hyperalgesia and motor improvement/worsening antagonized completely the effects of a conditioning procedure. We also measured the effects of opposing verbal suggestions on hormonal secretion and found that verbally induced expectations of increase/decrease of growth hormone (GH) and cortisol did not have any effect on the secretion of these hormones. However, if a preconditioning was performed with sumatriptan, a 5-HT(1B/1D) agonist that stimulates GH and inhibits cortisol secretion, a significant increase of GH and decrease of cortisol plasma concentrations were found after placebo administration, although opposite verbal suggestions were given. These findings indicate that verbally induced expectations have no effect on hormonal secretion, whereas they affect pain and motor performance. This suggests that placebo responses are mediated by conditioning when unconscious physiological functions such as hormonal secretion are involved, whereas they are mediated by expectation when conscious physiological processes such as pain and motor performance come into play, even though a conditioning procedure is performed. PMID- 12764121 TI - Morphine acutely regulates opioid receptor trafficking selectively in dendrites of nucleus accumbens neurons. AB - Morphine stimulates the internalization of mu-opioid receptors (MORs) in transfected cell models to a lesser degree than opioid peptides and other analgesic drugs, such as methadone, and previous studies have reported that morphine does not produce a detectable redistribution of MORs in neural tissue after either acute or chronic administration. Nevertheless, morphine produces profound physiological effects, raising the question of whether receptor trafficking plays any role in the in vivo actions of morphine. We investigated the effects of opiate drugs on recombinant and native opioid receptors in the nucleus accumbens, which plays an important role in mediating the behavioral effects of opiate drugs. Morphine and methadone differed in their effects on the internalization of epitope-tagged MORs in cell bodies, introduced by viral gene transfer and imaged by fluorescence microscopy. A mutation of the cytoplasmic tail that confers morphine-induced internalization in cultured cells had a similar effect on receptor trafficking in nucleus accumbens cell bodies. Surprisingly, in contrast to its failure to affect MOR distribution detectably in cell bodies, acute morphine administration produced a pronounced change in MOR distribution visualized in the processes of the same neurons. A similar effect of acute morphine administration was observed for endogenously expressed MORs by immunoelectron microscopy; the acute administration of morphine increased the density of MORs associated with internal membrane structures specifically in dendrites. These results provide the first evidence that morphine regulates the distribution of MORs in neuronal processes, suggesting that "compartment selective" membrane trafficking represents a previously unanticipated type of opioid receptor regulation contributing to the in vivo effects of opiate drugs on a physiologically relevant population of CNS neurons. PMID- 12764122 TI - Neuronal activity in the rostral superior colliculus related to the initiation of pursuit and saccadic eye movements. AB - The extinction of the central fixated stimulus before the appearance of a new target stimulus reduces the latency of saccades and pursuit, a phenomenon known as the "gap effect." The superior colliculus (SC) plays a prominent role in the gap effect for saccades, and recent data indicate that this structure also plays some role in the control of pursuit. We now show that the firing rate of buildup neurons in the rostral SC exhibits a gap effect during the initiation of both pursuit and saccadic eye movements to parafoveal targets. Most neurons exhibited an increase in tonic activity starting approximately 100 msec after the offset of the fixation spot, regardless of whether the target later appeared inside or outside of the response field of the neuron. The subsequent appearance of the target in the response field evoked phasic increases in activity that were approximately twice as large as the effects on tonic activity. For both pursuit and saccades, the levels of tonic and phasic activity were inversely correlated with latency on a trial-by-trial basis. These changes in activity provide a neuronal correlate for the shared effects on latency observed previously with the gap paradigm for pursuit and saccades. Finally, the phasic activity at pursuit onset exhibited a gap effect just like the target-evoked response, whereas the burst activity at saccade onset was fixed in amplitude. These results suggest how SC neurons may coordinate the initiation of pursuit and saccades: buildup activity may gate the initiation of pursuit, whereas it indirectly triggers saccades by recruiting a saccade-related burst. PMID- 12764123 TI - Selective hippocampal lesions do not increase adrenocortical activity. AB - It has been proposed that the hippocampus exerts a tonic inhibitory influence on the hypothalamic-pituitary-adrenal (HPA) stress axis. This claim rests, in particular, on the upregulation of corticosterone secretion and other measures of HPA activity after nonselective lesions of the hippocampal formation. We measured plasma corticosterone concentrations after selective neurotoxic damage to the hippocampus and the subiculum in rats. Concentrations were estimated during rest in the rat's home cage and at several time points after varying degrees of stress. Lesions of the hippocampus did not increase the concentration of corticosterone relative to control rats in any condition. Temporary inactivation of the hippocampus or the ventral subiculum by infusion of the GABAA receptor agonist muscimol also failed to induce hypersecretion, although hippocampal infusions did impair spatial memory. These results suggest that the hippocampus is not necessary for tonic inhibition of adrenocortical activity and imply that the HPA axis receives efficient negative feedback inhibition from other brain systems too. PMID- 12764124 TI - Histological and electrical properties of rat dorsal root ganglion neurons innervating the lower urinary tract. AB - We investigated whether primary afferent neurons innervating different regions of the lower urinary tract have different histochemical and electrophysiological properties. Neurons in rat L6-S1 DRG were identified by axonal transport of a fluorescent dye. Neurofilament-negative C-fiber cells comprise approximately 70% of bladder and proximal urethral afferent neurons that send axons through the pelvic nerves, but comprise a smaller proportion (51%) of distal urethral neurons that send axons through the pudendal nerves. Isolectin-B4 (IB4) binding was detected in a higher percentage (49%) of C-fiber neurons innervating the distal urethra than in those innervating the bladder or proximal urethra (18-22%). Neurofilament-positive A-fiber neurons innervating the distal urethra had a larger average somal size than neurons innervating the bladder or proximal urethra. In patch-clamp recordings, the majority (70%) of bladder and proximal urethral neurons were sensitive to capsaicin and exhibited TTX-resistant, high threshold action potentials, whereas a smaller proportion (53%) of distal urethral neurons exhibited TTX-resistant spikes. T-type Ca2+ currents were observed in 47% of distal urethral neurons with TTX-sensitive spikes, but not in TTX-sensitive bladder or proximal urethral neurons. In summary, afferent neurons innervating bladder or proximal urethra differ from those innervating distal urethra. The latter, which more closely resemble cutaneous afferent neurons, consist of a smaller number of C-fiber neurons containing a higher percentage of IB4-positive cells and a more diverse population of A-fiber neurons, some of which exhibit T-type Ca2+ channels. These differences may be related to different functions of respective target organs in the lower urinary tract. PMID- 12764125 TI - Recovery of amiloride-sensitive neural coding during regeneration of the gustatory nerve: behavioral-neural correlation of salt taste discrimination. AB - The chorda tympani (CT) nerve innervating the anterior tongue contains two types of NaCl-responsive fibers: one, the N-type, receives input from receptor cells, the NaCl responses of which are strongly inhibited by amiloride, whereas the other, the E-type, receives input from cells poorly sensitive or insensitive to amiloride. To investigate the formation of this differentially responsive neural system, we crushed the mouse CT nerve and examined the subsequent recovery of NaCl responses and amiloride sensitivity of the regenerated nerve and behavioral discrimination between NaCl and KCl. At 2 weeks after the nerve crush, no significant response of the nerve to chemical stimuli was observed. At 3 weeks, responses to salts gradually reappeared. In this period, almost all single fibers responding to NaCl were insensitive to amiloride (E-type). At 4 weeks, some of the single fibers showed amiloride sensitivity (N-type) and behavioral discrimination between NaCl and KCl reappeared. After >or=5 weeks, the number of N-type fibers had reached the control level and became approximately equal to that of E-type fibers. During the course of recovery, N-type and E-type fibers were clearly distinguishable on the basis of their amiloride sensitivities, their KCl/NaCl response ratios, and their concentration-response relationships to NaCl. These results suggest that two salt-responsive systems are independently reformed after the nerve crush. The selective synapse reformation may account for recovery of behavioral discrimination between NaCl and KCl after taste nerve crush and regeneration. It may also explain stable sensory coding for taste quality during the continuous turnover of receptor cells in the healthy animal. PMID- 12764126 TI - Step response analysis of thermotaxis in Caenorhabditis elegans. AB - The nematode Caenorhabditis elegans migrates toward a preferred temperature on a thermal gradient. A candidate neural network for thermotaxis in C. elegans has been identified, but the behavioral strategy implemented by this network is poorly understood. In this study, we tested whether thermal migration is achieved by modulating the probability of turning behavior, as in C. elegans chemotaxis. This was done by subjecting unrestrained wild-type, cryophilic, or thermophilic worms to rapid spatially uniform temperature steps (3 degrees C), up or down from the cultivation temperature. Each of the three types of worms we analyzed showed a different pair of responses to the two types of steps. Comparison of wild-type and mutant response patterns suggested a model in which thermal migration involves a unique response to the gradient depending on the orientation of the worm relative to its preferred temperature. Overall, however, turning probability was modulated in a manner consistent with a role for turning behavior in thermal migration. Our results suggest that sensory systems for thermotaxis and chemotaxis may converge on a common behavioral mechanism. PMID- 12764127 TI - Variable dopamine release probability and short-term plasticity between functional domains of the primate striatum. AB - Release of the neuromodulator dopamine (DA) is critical to the control of locomotion, motivation, and reward. However, the probability of DA release is not well understood. Current understanding of neurotransmitter release probability in the CNS is limited to the conventional synaptic amino acid transmitters (e.g., glutamate and GABA). These fast neurotransmitters are released with a repertoire of probabilities according to synapse type, and these probabilities show activity dependent plasticity according to synapse use. Synapses for neuromodulators such as DA, however, are designed for signaling that diverges temporally and spatially from that for fast neurotransmitters: DA receptors are exclusively metabotropic and at sites that extend to extrasynaptic locations and neighboring synapses. In this study, the release probability of DA was explored in real time in limbicversus motor-associated functional domains of the striatum of a primate (marmoset; Callithrix jacchus) using fast-scan voltammetry at a carbon-fiber microelectrode. We show that the probability of axonal DA release varies with striatal domain. Furthermore, release probability exhibits a short-term, activity dependent plasticity that ranges from depression to facilitation in motor-through limbic-associated regions, respectively. Rapid plasticity does not result from metabotropic D2-like DA receptor activation or ionotropic GABA(A) receptor effects but is dependent on Ca2+ availability. These data reveal that rapid dynamics in DA release probability will participate in the transmission of the patterns and frequencies encoded by DA neuron action potential discharge. Furthermore, the regional variation in these features indicates that limbic versus motor-associated DA neurons are permitted to generate diverse DA signals in response to a given firing pattern. PMID- 12764128 TI - Social influences on the arginine vasotocin system are independent of gonads in a sex-changing fish. AB - Many neuropeptide systems subserving sex-typical behavior are dependent on sex steroids for both their organization early in life and activation during maturity. The arginine vasopressin/vasotocin (AVP/AVT) system is strongly androgen dependent in many species and critically mediates responses to sociosexual stimuli. The bluehead wrasse is a teleost fish that exhibits a female to-male sex change in response to social cues, and neither the development nor the maintenance of male-typical behavior depends on the presence of gonads. To examine social and gonadal inputs on the AVP/AVT system in the preoptic area (POA) of the hypothalamus, we conducted three field experiments. In the first experiment, we found that AVT mRNA abundance is higher in sex-changing females that attain social dominance and display dominant male behavior than in subordinate females, regardless of whether the dominant females were intact or ovariectomized. However, AVT-immunoreactive (IR) soma size in the gigantocellular POA (gPOA), but not in the magnocellular or parvocellular POA, increased only when females were displaying both dominant male behavior and had developed testes. In the second experiment, castration of dominant terminal-phase males had no effect on AVT mRNA abundance or any behavior we measured but did increase gPOA AVT-IR soma size compared with sham-operated controls. In the third experiment, 11-ketotestosterone implants in socially subordinate, ovariectomized females had no effect on either AVT mRNA abundance or AVT-IR soma size compared with controls. These results demonstrate that the AVT neural phenotype in the bluehead wrasse can be strongly influenced by social status, and that these social influences can be manifested independent of gonads. PMID- 12764129 TI - SUMO modification of STAT1 and its role in PIAS-mediated inhibition of gene activation. AB - The PIAS (protein inhibitors of activated STAT) family of proteins were first discovered as inhibitors of activated signal transducers and activators of transcription (STATs). More recently these proteins have been shown to function as E3 ligases that promote the SUMO modification of a number of transcription regulators. We have investigated the relationship between the effects of PIAS proteins on STAT1 transcriptional activity and the ability of the PIAS proteins to function as SUMO E3 ligases. We demonstrate that STAT1 is a substrate for SUMO modification and that PIASx-alpha, but not PIAS1, functions as an E3 ligase to promote STAT1 modification. In addition, we have mapped the major site for SUMO modification on STAT1 to lysine 703. This lysine residue is in close proximity to the regulatory tyrosine residue at position 701, whose phosphorylation mediates STAT1 activation in response to cytokine signaling. Mutation of lysine 703 to arginine abolishes SUMO modification of STAT1 both in vitro and in vivo. However, this mutation does not affect the activation of STAT1 or the ability of either PIAS1 or PIASx-alpha to function as an inhibitor of STAT1-mediated transcription activation. Our findings demonstrate that inhibition of STAT1 by PIAS proteins does not require SUMO modification of STAT1 itself. SUMO modification of STAT1 may nonetheless be functionally important given the close proximity between the SUMO modification site and tyrosine 701. PMID- 12764130 TI - The kelch repeat protein, Tea1, is a potential substrate target of the p21 activated kinase, Shk1, in the fission yeast, Schizosaccharomyces pombe. AB - The p21-activated kinase (PAK) homolog, Shk1, is a critical component of a multifunctional Ras/Cdc42/PAK complex required for viability, polarized growth and cell shape, and sexual differentiation in the fission yeast, Schizosaccharomyces pombe. Substrate targets of the Shk1 kinase have not previously been described. Here we show that the S. pombe cell polarity factor, Tea1, is directly phosphorylated by Shk1 in vitro. We demonstrate further that Tea1 is phosphorylated in S. pombe cells and that its level of phosphorylation is significantly reduced in cells defective in Shk1 function. Consistent with a role for Tea1 as a potential downstream effector of Shk1, we show that a tea1 null mutation rescues the Shk1 hyperactivity-induced lethal phenotype caused by loss of function of the essential Shk1 inhibitor, Skb15. All phenotypes associated with Skb15 loss, including defects in actin cytoskeletal organization, chromosome segregation, and cytokinesis, are suppressed by tea1 Delta, suggesting that Tea1 is a potential mediator of multiple Shk1 functions. S. pombe cells carrying a weak hypomorphic allele of shk1 together with a tea1 Delta mutation exhibit a cytokinesis defective phenotype that is significantly more severe than that observed in the respective single mutants, providing evidence that Shk1 and Tea1 cooperate to regulate cytokinesis. In addition, we show that S. pombe cells carrying the orb2-34 allele of shk1 exhibit a pattern of monopolar growth similar to that observed in tea1 Delta cells, suggesting that Shk1 and Tea1 may regulate one or more common processes involved in the regulation of polarized cell growth. Taken together, our results strongly implicate Tea1 as a potential substrate effector of the Shk1 kinase. PMID- 12764131 TI - Membrane topology of a metabotropic glutamate receptor. AB - The metabotropic glutamate receptors (mGluRs) have been predicted to have a classical seven transmembrane domain structure similar to that seen for members of the G-protein-coupled receptor (GPCR) superfamily. However, the mGluRs (and other members of the family C GPCRs) show no sequence homology to the rhodopsin like GPCRs, for which this seven transmembrane domain structure has been experimentally confirmed. Furthermore, several transmembrane domain prediction algorithms suggest that the mGluRs have a topology that is distinct from these receptors. In the present study, we set out to test whether mGluR5 has seven true transmembrane domains. Using a variety of approaches in both prokaryotic and eukaryotic systems, our data provide strong support for the proposed seven transmembrane domain model of mGluR5. We propose that this membrane topology can be extended to all members of the family C GPCRs. PMID- 12764132 TI - The reverse activity of human acid ceramidase. AB - An overexpression system was recently developed to produce and purify recombinant, human acid ceramidase. In addition to ceramide hydrolysis, the purified enzyme was able to catalyze ceramide synthesis using [14C]lauric acid and sphingosine as substrates. Herein we report detailed characterization of this acid ceramidase-associated "reverse activity" and provide evidence that this reaction occurs in situ as well as in vitro. The pH optimum of the reverse reaction was approximately 5.5, as compared with approximately 4.5 for the hydrolysis reaction. Non-ionic detergents and zinc cations inhibited the activity, whereas most other cations were stimulatory. Of note, sphingomyelin also was very inhibitory toward this reaction, whereas the anionic lipids, phosphatidic acid and phosphatidylserine, were stimulatory. Of various sphingosine stereoisomers tested in the reverse reaction, only the natural, D erythro form could efficiently serve as a substrate. Using D-erythro-sphingosine and lauric acid as substrates, the reaction followed normal Michaelis-Menten kinetics. The Km and Vmax values toward sphingosine were 23.75 microM and 208.3 pmol/microg/h, respectively, whereas for lauric acid they were 73.76 microM and 232.5 pmol/microg/h, respectively. Importantly, the reverse activity was reduced in cell lysates from a Farber disease patient to the same extent as the acid ceramidase activity. Furthermore, when 12-(N-methyl-N-(7-nitrobenz-2-oxa-1,3 diazol-4-yl)) (NBD)-conjugated lauric acid and sphingosine were added to cultured lymphoblasts from a Farber disease patient in the presence of fumonisin B (1), the conversion to NBD-ceramide was reduced approximately 30% when compared with normal cells. These data provide important new information on human acid ceramidase and further document its central role in sphingolipid metabolism. PMID- 12764133 TI - Putative zinc-sensing zinc fingers of metal-response element-binding transcription factor-1 stabilize a metal-dependent chromatin complex on the endogenous metallothionein-I promoter. AB - The metalloregulatory functions of metal-response element-binding transcription factor-1 (MTF-1) have been mapped, in part, to its six highly conserved zinc fingers. Here we examined the ability of zinc finger deletion mutants of mouse MTF-1 to regulate the endogenous metallothionein-I (MT-I) gene in cells lacking endogenous MTF-1. MTF-1 knockout mouse embryo fibroblasts were transfected with expression vectors for FLAG-tagged MTF-1 (MTF-1flag) or finger deletion mutants of MTF-1flag and then assayed for metal induction of MT-I gene expression, nuclear translocation, and in vitro DNA-binding activity of MTF-1 and its stable association with the endogenous chromosomal MT-I promoter. Intact MTF-1flag restored metal responsiveness of the MT-I gene, underwent nuclear translocation, displayed increased in vitro DNA binding in response to zinc and less so to cadmium, and rapidly formed a stable complex with the MT-I promoter chromatin in response to both of these metals. In contrast, although deletion of finger 1, fingers 5 and 6, or finger 6 only had variable effects on the nuclear localization and in vitro DNA-binding activity of MTF-1, each of these finger deletion mutants severely attenuated metal-induced MTF-1 binding to the MT-I promoter chromatin and activation of the endogenous MT-I gene. These results demonstrated that the metal-induced recruitment of MTF-1 to the MT-I promoter is a rate-limiting step in its metalloregulatory function and that an intact zinc finger domain is required for this recruitment. During the course of these studies, it was discovered that mouse MTF-1 is polymorphic. The impact of these polymorphisms on MTF-1 metalloregulatory functions is discussed. PMID- 12764134 TI - Autophosphorylation of cGMP-dependent protein kinase type II. AB - Cyclic nucleotides are shown to stimulate the autophosphorylation of type II cGMP dependent protein kinase (cGK) on multiple sites. Mass spectrometric based analyses, using a quadrupole time-of-flight-mass spectrometry instrument revealed that cGMP stimulated the in vitro phosphorylation of residues Ser110 and Ser114, and, at a slow rate, of Ser126 and Thr109 or Ser117, all located in the autoinhibitory region. In addition Ser445 was found to be phosphorylated in a cGMP-dependent manner, whereas Ser110 and Ser97 were already prephosphorylated to a large extent in Sf9 cells. cGMP-dependent phosphorylation of cGK II was also demonstrated in intact COS-1 cells and intestinal epithelium. Substitution of most of the potentially autophosphorylated residues for alanines largely abolished the cGMP stimulation of the autophosphorylation. Prolonged autophosphorylation of purified recombinant cGK II in vitro resulted in a 40-50% increase in basal kinase activity, but its maximal cGMP-stimulated activity and the EC50 for cGMP remained unaltered. Mutation of the major phosphorylatable serines 110, 114, and 445 into "phosphorylation-mimicking" glutamates had no effect on the kinetic parameters of cGK II. However, replacing the slowly autophosphorylated residue Ser126 by Glu rendered cGK II constitutively active. These results show that the fast phase of cyclic nucleotide-stimulated autophosphorylation of cGK II has a relatively small feed forward effect on its activity, whereas the secondary phase, presumably involving Ser126 phosphorylation, may generate a constitutively active form of the enzyme. PMID- 12764135 TI - The transforming activity of Ski and SnoN is dependent on their ability to repress the activity of Smad proteins. AB - The regulation of cell growth and differentiation by transforming growth factor beta (TGF-beta) is mediated by the Smad proteins. In the nucleus, the Smad proteins are negatively regulated by two closely related nuclear proto oncoproteins, Ski and SnoN. When overexpressed, Ski and SnoN induce oncogenic transformation of chicken embryo fibroblasts. However, the mechanism of transformation by Ski and SnoN has not been defined. We have previously reported that Ski and SnoN interact directly with Smad2, Smad3, and Smad4 and repress their ability to activate TGF-beta target genes through multiple mechanisms. Because Smad proteins are tumor suppressors, we hypothesized that the ability of Ski and SnoN to inactivate Smad function may be responsible for their transforming activity. Here, we show that the receptor regulated Smad proteins (Smad2 and Smad3) and common mediator Smad (Smad4) bind to different regions in Ski and SnoN. Mutation of both regions, but not each region alone, markedly impaired the ability of Ski and SnoN to repress TGF-beta-induced transcriptional activation and cell cycle arrest. Moreover, when expressed in chicken embryo fibroblasts, mutant Ski or SnoN defective in binding to the Smad proteins failed to induce oncogenic transformation. These results suggest that the ability of Ski and SnoN to repress the growth inhibitory function of the Smad proteins is required for their transforming activity. This may account for the resistance to TGF-beta-induced growth arrest in some human cancer cell lines that express high levels of Ski or SnoN. PMID- 12764136 TI - 14-3-3 acts as an intramolecular bridge to regulate cdc25B localization and activity. AB - One of the major regulators of mitosis in somatic cells is cdc25B. cdc25B is tightly regulated at multiple levels. The final activation step involves the regulated binding of 14-3-3 proteins. Previous studies have demonstrated that Ser 323 is a primary 14-3-3 binding site in cdc25B, which influences its activity and cellular localization. 14-3-3 binding to this site appeared to interact with the N-terminal domain of cdc25B to regulate its activity. The presence of consensus 14-3-3 binding sites in the N-terminal domain suggested that the interaction is through direct binding of the 14-3-3 dimer to sites in the N-terminal domain. We have identified Ser-151 and Ser-230 in the N-terminal domain as functional 14-3-3 binding sites utilized by cdc25B in vivo. These low affinity sites cooperate to bind the 14-3-3 dimer bound to the high affinity Ser-323 site, thus forming an intramolecular bridge that constrains cdc25B structure to prevent access of the catalytic site. Loss of 14-3-3 binding to either N-terminal site relaxes cdc25B structure sufficiently to permit access to the catalytic site, and the nuclear export sequence located in the N-terminal domain. Mutation of the Ser-323 site was functionally equivalent to the mutation of all three sites, resulting in the complete loss of 14-3-3 binding, increased access of the catalytic site, and access to nuclear localization sequence. PMID- 12764138 TI - The negative transcriptional regulator NmrA discriminates between oxidized and reduced dinucleotides. AB - NmrA, a transcription repressor involved in the regulation of nitrogen metabolism in Aspergillus nidulans,is a member of the short-chain dehydrogenase reductase superfamily. Isothermal titration calorimetry and differential scanning calorimetry have been used to show NmrA binds NAD+ and NADP+ with similar affinity (average KD 65 microM) but has a greatly reduced affinity for NADH and NADPH (average KD 6.0 mM). The structure of NmrA in a complex with NADP+ reveals how repositioning a His-37 side chain allows the different conformations of NAD+ and NADP+ to be accommodated. Modeling NAD(P)H into NmrA indicated that steric clashes, attenuation of electrostatic interactions, and loss of aromatic ring stacking can explain the differing affinities of NAD(P)+/NAD(P)H. The ability of NmrA to discriminate between the oxidized and reduced forms of the dinucleotides may be linked to a possible role in redox sensing. Isothermal titration calorimetry demonstrated that NmrA and a C-terminal fragment of the GATA transcription factor AreA interacted with a 1:1 stoichiometry and an apparent KD of 0.26 microM. NmrA was unable to bind the nitrogen metabolite repression signaling molecules ammonium or glutamine. PMID- 12764137 TI - MRP8, ATP-binding cassette C11 (ABCC11), is a cyclic nucleotide efflux pump and a resistance factor for fluoropyrimidines 2',3'-dideoxycytidine and 9'-(2' phosphonylmethoxyethyl)adenine. AB - MRP8 (ABCC11) is a recently identified cDNA that has been assigned to the multidrug resistance-associated protein (MRP) family of ATP-binding cassette transporters, but its functional characteristics have not been determined. Here we examine the functional properties of the protein using transfected LLC-PK1 cells. It is shown that ectopic expression of MRP8 reduces basal intracellular levels of cAMP and cGMP and enhances cellular extrusion of cyclic nucleotides in the presence or absence of stimulation with forskolin or SIN-1A. Analysis of the sensitivity of MRP8-overexpressing cells revealed that they are resistant to a range of clinically relevant nucleotide analogs, including the anticancer fluoropyrimidines 5'-fluorouracil (approximately 3-fold), 5'-fluoro-2' deoxyuridine (approximately 5-fold), and 5'-fluoro-5'-deoxyuridine (approximately 3-fold), the anti-human immunodeficiency virus agent 2',3'-dideoxycytidine (approximately 6-fold) and the anti-hepatitis B agent 9'-(2' phosphonylmethoxynyl)adenine (PMEA) (approximately 5-fold). By contrast, increased resistance was not observed for several natural product chemotherapeutic agents. In accord with the notion that MRP8 functions as a drug efflux pump for nucleotide analogs, MRP8-transfected cells exhibited reduced accumulation and increased efflux of radiolabeled PMEA. In addition, it is shown by the use of in vitro transport assays that MRP8 is able to confer resistance to fluoropyrimidines by mediating the MgATP-dependent transport of 5'-fluoro-2' deoxyuridine monophosphate, the cytotoxic intracellular metabolite of this class of agents, but not of 5'-fluorouracil or 5'-fluoro-2'-deoxyuridine. We conclude that MRP8 is an amphipathic anion transporter that is able to efflux cAMP and cGMP and to function as a resistance factor for commonly employed purine and pyrimidine nucleotide analogs. PMID- 12764140 TI - Transcriptional elongation by RNA polymerase II and histone methylation. AB - mRNA synthesis in eukaryotic organisms is a key biological process that is regulated at multiple levels. From the covalent modifications of chromatin by a number of chromatin remodeling complexes during the initiation and activation steps of transcription to the processing of mRNA transcripts, a very large consortium of proteins and multiprotein complexes is critical for gene expression by RNA polymerase II. The list of proteins essential for the successful synthesis of mRNA continues to grow at a rapid pace. Recent advances in this area of research have been focused on transcription through chromatin. In this article, we will review the recent literature linking the key biochemical process of transcriptional elongation by RNA polymerase II to histone methylation by COMPASS, Dot1p, and Set2 methyltransferases. PMID- 12764139 TI - Calpain regulates enterocyte brush border actin assembly and pathogenic Escherichia coli-mediated effacement. AB - This study identifies calpain as being instrumental for brush border (BB) microvillus assembly during differentiation and effacement during bacterial pathogenesis. Calpain activity is decreased by 25-80% in Caco 2 lines stably overexpressing calpastatin, the physiological inhibitor of calpain, and the effect is proportional to the calpastatin/calpain ratio. These lines exhibit a 2.5-fold reduction in the rate of microvillus extension. Apical microvillus assembly is reduced by up to 50%, as measured by quantitative fluorometric microscopy (QFM) of ezrin, indicating that calpain recruits ezrin to BB microvilli. Calpain inhibitors ZLLYCHN2, MDL 28170, and PD 150606 block BB assembly and ezrin recruitment to the BB. The HIV protease inhibitor ritonavir, which inhibits calpain at clinically relevant concentrations, also blocks BB assembly, whereas cathepsin and proteasome inhibitors do not. Microvillus effacement is inhibited after exposure of calpastatin-overexpressing cells to enteropathogenic Escherichia coli. These results suggest that calpain regulates BB assembly as well as pathological effacement, and indicate that it is an important regulator involved in HIV protease inhibitor toxicity and host microbial pathogen interactions. PMID- 12764141 TI - Three-dimensional structural interactions of insulin and its receptor. PMID- 12764142 TI - Calcium-triggered membrane fusion proceeds independently of specific presynaptic proteins. AB - Complexes of specific presynaptic proteins have been hypothesized to drive or catalyze the membrane fusion steps of exocytosis. Here we use a stage-specific preparation to test the roles of SNAREs, synaptotagmin, and SNARE-binding proteins in the mechanism of Ca2+-triggered membrane fusion. Excess exogenous proteins, sufficient to block SNARE interactions, did not inhibit either the Ca2+ sensitivity, extent, or kinetics of fusion. In contrast, despite a limited effect on SNARE and synaptotagmin densities, treatments with high doses of chymotrypsin markedly inhibited fusion. Conversely, low doses of chymotrypsin had no effect on the Ca2+ sensitivity or extent of fusion but did alter the kinetic profile, indicating a more direct involvement of other proteins in the triggered fusion pathway. SNAREs, synaptotagmin, and their immediate binding partners are critical to exocytosis at a stage other than membrane fusion, although they may still influence the triggered steps. PMID- 12764143 TI - Regulation of hypoxia-inducible factor-1alpha protein level during hypoxic conditions by the phosphatidylinositol 3-kinase/Akt/glycogen synthase kinase 3beta pathway in HepG2 cells. AB - Hypoxia initiates an intracellular signaling pathway leading to the activation of the transcription factor hypoxia-inducible factor-1 (HIF-1). HIF-1 activity is regulated through different mechanisms involving stabilization of HIF-1alpha, phosphorylations, modifications of redox conditions, and interactions with coactivators. However, it appears that some of these steps can be cell type specific. Among them, the involvement of the phosphatidylinositol 3-kinase (PI3K)/Akt pathway in the regulation of HIF-1 by hypoxia remains controversial. Here, we investigated the activation state of PI3K/Akt/glycogen synthase kinase 3beta (GSK3beta) in HepG2 cells. Increasing incubation times in hypoxia dramatically decreased both the phosphorylation of Akt and the inhibiting phosphorylation of GSK3beta. The PI3K/Akt pathway was necessary for HIF-1alpha stabilization early during hypoxia. Indeed, its inhibition was sufficient to decrease HIF-1alpha protein level after 5-h incubation in hypoxia. However, longer exposure (16 h) in hypoxia resulted in a decreased HIF-1alpha protein level compared with early exposure (5 h). At that time, Akt was no longer present or active, which resulted in a decrease in the inhibiting phosphorylation of GSK3beta on Ser-9 and hence in an increased GSK3beta activity. GSK3 inhibition reverted the effect of prolonged hypoxia on HIF-1alpha protein level; more stabilized HIF-1alpha was observed as well as increased HIF-1 transcriptional activity. Thus, a prolonged hypoxia activates GSK3beta, which results in decreased HIF-1alpha accumulation. In conclusion, hypoxia induced a biphasic effect on HIF-1alpha stabilization with accumulation in early hypoxia, which depends on an active PI3K/Akt pathway and an inactive GSK3beta, whereas prolonged hypoxia results in the inactivation of Akt and activation of GSK3beta, which then down-regulates the HIF-1 activity through down-regulation of HIF-1alpha accumulation. PMID- 12764144 TI - Acetyl-CoA carboxylase beta gene is regulated by sterol regulatory element binding protein-1 in liver. AB - Acetyl-CoA carboxylase (ACC) exists as two major isoforms originated from separate genes: ACCalpha (or ACC1) and ACCbeta (or ACC2). Previous data revealed that ACCbeta has two forms of mRNA with different 5'-untranslated regions derived by different usage of promoters, I and II, in human. In this study, we revealed that ACCbeta expression in liver is markedly stimulated by food intake at the transcriptional level. In the process of this induction in rat liver, promoter II plays the major role in regulating the expression of ACCbeta gene. The transient transfection with promoter II-luciferase reporters elucidated that the region from -93 to -38 nucleotides is important for the responsiveness to sterol regulatory element-binding protein-1 (SREBP-1), which is known to be the principle mediator for the stimulation of gene transcriptions by insulin and diet. The Sp1-binding site (-71 to -66) and neighboring two conserved SREs (-62 to -44) play a critical role in the stimulation of ACCbeta gene expression by SREBP-1. In vivo chromatin immunoprecipitation assay revealed that SREBP-1 directly bound to ACCbeta promoter II in liver, and its binding was regulated by the diet. This study provides evidence that ACCbeta expression in liver is regulated at the transcriptional level by the direct interaction of SREBP-1 with promoter II. PMID- 12764146 TI - A new subunit of the epithelial Na+ channel identifies regions involved in Na+ self-inhibition. AB - The epithelial Na+ channel (ENaC) is the apical entry pathway for Na+ in many Na+ reabsorbing epithelia. ENaC is a heterotetrameric protein composed of homologous alpha, beta, and gamma subunits. Mutations in ENaC cause severe hypertension or salt wasting in humans; and consequently, ENaC activity is tightly controlled. According to the concept of Na+ self-inhibition, the extracellular Na+ ion itself can reduce ENaC activity. The molecular basis for Na+ self-inhibition is unknown. Here, we describe cloning of a new ENaC subunit from Xenopus laevis (epsilonxENaC). epsilonxENaC can replace alphaxENaC and formed functional, highly selective, amiloride-sensitive Na+ channels when coexpressed with betaxENaC and gammaxENaC. Channels containing epsilonxENaC showed strong inhibition by extracellular Na+. This Na+ self-inhibition was significantly slower than for alphaxENaC-containing channels. Using site-directed mutagenesis, we show that the proximal part of the large extracellular domain controls the speed of self inhibition. This suggests that this region is involved in conformational changes during Na+ self-inhibition. PMID- 12764145 TI - Dependence of mu-conotoxin block of sodium channels on ionic strength but not on the permeating [Na+]: implications for the distinctive mechanistic interactions between Na+ and K+ channel pore-blocking toxins and their molecular targets. AB - Mu-conotoxins (mu-CTXs) are Na+ channel-blocking, 22-amino acid peptides produced by the sea snail Conus geographus. Although K+ channel pore-blocking toxins show specific interactions with permeant ions and strong dependence on the ionic strength (mu), no such dependence has been reported for mu-CTX and Na+ channels. Such properties would offer insight into the binding and blocking mechanism of mu CTX as well as functional and structural properties of the Na+ channel pore. Here we studied the effects of mu and permeant ion concentration ([Na+]) on mu-CTX block of rat skeletal muscle (mu1, Nav1.4) Na+ channels. Mu-CTX sensitivity of wild-type and E758Q channels increased significantly (by approximately 20-fold) when mu was lowered by substituting external Na+ with equimolar sucrose (from 140 to 35 mm Na+); however, toxin block was unaltered (p > 0.05) when mu was maintained by replacement of [Na+] with N-methyl-d-glucamine (NMG+), suggesting that the enhanced sensitivity at low mu was not due to reduction in [Na+]. Single channel recordings identified the association rate constant, k(on), as the primary determinant of the changes in affinity (k(on) increased 40- and 333-fold for mu-CTX D2N/R13Q and D12N/R13Q, respectively, when symmetric 200 mm Na+ was reduced to 50 mm). In contrast, dissociation rates changed <2-fold for the same derivatives under the same conditions. Experiments with additional mu-CTX derivatives identified toxin residues Arg-1, Arg-13, and Lys-16 as important contributors to the sensitivity to external mu. Taken together, our findings indicate that mu-CTX block of Na+ channels depends critically on mu but not specifically on [Na+], contrasting with the known behavior of pore-blocking K+ channel toxins. These findings suggest that different degrees of ion interaction, underlying the fundamental conduction mechanisms of Na+ and K+ channels, are mirrored in ion interactions with pore-blocking toxins. PMID- 12764148 TI - Increased hepatic fructose 2,6-bisphosphate after an oral glucose load does not affect gluconeogenesis. AB - The generally accepted metabolic concept that fructose 2,6-bisphosphate (Fru-2,6 P2) inhibits gluconeogenesis by directly inhibiting fructose 1,6-bisphosphatase is based entirely on in vitro observations. To establish whether gluconeogenesis is indeed inhibited by Fru-2,6-P2 in intact animals, a novel NMR method was developed using [U-13C]glucose and 2H2O as tracers. The method was used to estimate the sources of plasma glucose from gastric absorption of oral [U 13C]glucose, from gluconeogenesis, and from glycogen in 24-h fasted rats. Liver Fru-2,6-P2 increased approximately 10-fold shortly after the glucose load, reached a maximum at 60 min, and then dropped to base-line levels by 150 min. The gastric contribution to plasma glucose reached approximately 50% at 30 min after the glucose load and gradually decreased thereafter. Although the contribution of glycogen to plasma glucose was small, glucose formed from gluconeogenesis was substantial throughout the study period even when liver Fru-2,6-P2 was high. Liver glycogen repletion was also brisk throughout the study period, reaching approximately 30 micromol/g at 3 h. These data demonstrate that Fru-2,6-P2 does not inhibit gluconeogenesis significantly in vivo. PMID- 12764150 TI - Functional characterization of the propeptide of Plasmodium falciparum subtilisin like protease-1. AB - Erythrocyte invasion by the malaria merozoite is prevented by serine protease inhibitors. Various aspects of the biology of Plasmodium falciparum subtilisin like protease-1 (PfSUB-1), including the timing of its expression and its apical location in the merozoite, suggest that this enzyme is involved in invasion. Recombinant PfSUB-1 expressed in a baculovirus system is secreted in the p54 form, noncovalently bound to its cognate propeptide, p31. To understand the role of p31 in PfSUB-1 maturation, we examined interactions between p31 and both recombinant and native enzymes. CD analyses revealed that recombinant p31 (rp31) possesses significant secondary structure on its own, comparable with that of folded propeptides of some bacterial subtilisins. Kinetic studies demonstrated that rp31 is a fast binding, high affinity inhibitor of PfSUB-1. Inhibition of two bacterial subtilisins by rp31 was much less effective, with inhibition constants 49-60-fold higher than that for PfSUB-1. Single (at the P4 or P1 position) or double (at P4 and P1 positions) point mutations of residues within the C-terminal region of rp31 had little effect on its inhibitory activity, and truncation of 11 residues from the rp31 C terminus substantially reduced, but did not abolish, inhibition. None of these modifications prevented binding to the PfSUB-1 catalytic domain or rendered the propeptide susceptible to proteolytic digestion by PfSUB-1. These studies provide new insights into the function of the propeptide in PfSUB-1 activation and shed light on the structural requirements for interaction with the catalytic domain. PMID- 12764149 TI - Methenyltetrahydrofolate synthetase regulates folate turnover and accumulation. AB - Cellular folate deficiency impairs one-carbon metabolism, resulting in decreased fidelity of DNA synthesis and inhibition of numerous S-adenosylmethionine dependent methylation reactions including protein and DNA methylation. Cellular folate concentrations are influenced by folate availability, cellular folate transport efficiency, folate polyglutamylation, and folate turnover specifically through degradation. Folate cofactors are highly susceptible to oxidative degradation in vitro with the exception of 5-formyltetrahydrofolate, which may be a storage form of folate. In this study, we determined the effects of depleting cytoplasmic 5-formyltetrahydrofolate on cellular folate concentrations and folate turnover rates in cell cultures by expressing the human methenyltetrahydrofolate synthetase cDNA in human MCF-7 cells and SH-SY5Y neuroblastoma. Cells with increased methenyltetrahydrofolate synthetase activity exhibited: 1) increased rates of folate turnover, 2) elevated generation of p-aminobenzoylglutamate in culture medium, 3) depressed cellular folate concentrations independent of medium folic acid concentrations, and 4) increased average polyglutamate chain lengths of folate cofactors. These data indicate that folate catabolism and folate polyglutamylation are competitive reactions that influence cellular folate concentrations, and that increased methenyltetrahydrofolate synthetase activity accelerates folate turnover rates, depletes cellular folate concentrations, and may account in part for tissue-specific differences in folate accumulation. PMID- 12764151 TI - Src family kinase-independent signal transduction and gene induction by leukemia inhibitory factor. AB - Members of the interleukin-6 (IL-6) family of cytokines exert their biological effects via binding to their cognate ligand-binding receptor subunit on a target cell. The subsequent recruitment of the common signal transducer glycoprotein 130 and activation of the JAK/STAT and SHP-2/Ras/mitogen-activated protein kinase (MAPK) pathways are responsible for the majority of cellular responses elicited by IL-6 cytokines. Several types of experiments suggest that the Src family of kinases (SFK) also participates in IL-6 family cytokine-mediated signaling events. SYF cells, which lack expression of SFKs Src, Yes, and Fyn, were used to determine the role of SFKs in IL-6 family cytokine signaling and gene induction. SYF and wild type (WT) control fibroblasts displayed similar activation of signaling intermediates following stimulation with leukemia inhibitory factor (LIF). LIF-stimulated tyrosine phosphorylation of SHP-2 and subsequent activation of MAPK in SYF cells were identical to that seen in LIF-stimulated WT cells. Both LIF-stimulated tyrosine phosphorylation of STAT1 and STAT3, as well as LIF stimulated DNA binding activity of STAT-containing nuclear complexes were indistinguishable when compared in SYF and WT cells. In addition, the phosphatidylinositol 3-kinase-sensitive Akt kinase and p38 MAPK were activated by LIF in both SYF and WT cells. Furthermore, LIF-stimulated expression of c-fos, egr-1, and suppressor of cytokine signaling-3 was retained in SYF cells. The IL-6 family cytokine oncostatin M was also capable of activating MAPK, STAT3, STAT1, Akt, and p38 in both WT and SYF cells. These results demonstrate that IL-6 family cytokines can activate a full repertoire of signaling pathways and induce gene expression independent of SFKs. PMID- 12764153 TI - Characterization of a myeloid tyrosine phosphatase, Lyp, and its role in the Bcr Abl signal transduction pathway. AB - The Bcr-Abl protein-tyrosine kinase is implicated in the development of chronic myeloid leukemia. The potential role of protein-tyrosine phosphatase in the regulation of Bcr-Abl signaling was explored. First, expression patterns of tyrosine phosphatases in leukemic cell lines were investigated using degenerate primers for reverse transcription-PCR followed by cloning and sequencing of the cDNA. Distinct patterns of distribution of phosphatase were found in erythroid and myeloid leukemic cell lines. Whereas some phosphatases were ubiquitously expressed, others were limited to specific cell types. Surprisingly, a previously cloned "lymphocyte-specific" phosphatase, Lyp, was frequently detected in a number of myeloid cell lines as well as normal granulocytes and monocytes. Lyp was localized to the cytosol, and overexpression of Lyp caused reduction in the phosphorylation levels of multiple proteins in KCL22 chronic myeloid leukemia blast cells including Cbl, Bcr-Abl, Erk1/2, and CrkL. Co-expression of Lyp and Bcr-Abl in Cos-7 cells resulted in decreased levels of Bcr-Abl, Grb2, and Myc. Overexpression of Lyp markedly suppressed anchorage-independent clonal growth of KCL22 cells. Taken together, the data suggest that Lyp may play an antagonistic role in signaling by the Bcr-Abl fusion protein. PMID- 12764152 TI - Identification of phosphorylation sites in AMP-activated protein kinase (AMPK) for upstream AMPK kinases and study of their roles by site-directed mutagenesis. AB - Bacterially expressed heterotrimeric (alpha1, beta1, and gamma1) wild-type, catalytically inactive, and constitutively active forms of AMP-activated protein kinase (AMPK) were used to study phosphorylation by an upstream AMPK kinase preparation. Here, we report the identification of two new phosphorylation sites in the alpha-subunit, viz. Thr258 and Ser485 (Ser491 in the alpha2-subunit) by mass spectrometry, in addition to the previously characterized Thr172 site. Also, autophosphorylation sites in the beta1-subunit were identified as Ser96, Ser101, and Ser108. Mutagenesis of Thr172, Thr258, and Ser485 to acidic residues to mimic phosphorylation in the recombinant proteins indicated that Thr172 was involved in AMPK activation, whereas Thr258 and Ser485 were not. Transfection of the non phosphorylatable S485A and T258A mutants in CCL13 cells subjected to stresses known to activate AMPK either by increasing the AMP:ATP ratio (slow lysis) or without changing adenine nucleotide concentrations (hyperosmolarity) resulted in no significant differences in AMPK activation. All three sites within the alpha subunit were phosphorylated in vivo, as seen in AMPK immunoprecipitated from anoxic rat liver. In transfected CCL13 cells, the level of Ser485 phosphorylation did not change upon AMPK activation. The newly identified phosphorylation sites could play a subtle role in the regulation of AMPK, e.g. in subcellular localization or substrate recognition. PMID- 12764154 TI - Activation of store-operated calcium channels: assessment of the role of snare mediated vesicular transport. AB - Store-operated calcium channels (SOC) play a central role in cellular calcium homeostasis. Although it is well established that SOC are activated by depletion of the endoplasmic reticulum calcium stores, the molecular mechanism underlying this effect remains ill defined. It has been suggested that SOC activation requires fusion of endomembrane vesicles with the plasmalemma. In this model, SNARE-dependent exocytosis is proposed to deliver channels or their activators to the surface membrane to initiate calcium influx. To test this hypothesis, we studied the requirement for membrane fusion events in SOC activation, using a variety of dominant-negative constructs and toxins that interfere with SNARE function. Botulinum neurotoxin A (BotA), which cleaves SNAP-25, did not prevent SOC activation. Moreover, SNAP-25 was not detectable in the cells where BotA was reported earlier to inhibit SOC. Instead, the BotA-insensitive SNAP-23 was present. Impairment of VAMP function was similarly without effect on SOC opening. We also tested the role of N-ethylmaleimide-sensitive factor, a global regulator of SNARE-mediated membrane fusion. Expression of a mutated N-ethylmaleimide sensitive factor construct inhibited all aspects of membrane traffic tested, including recycling of transferrin receptors to the plasma membrane, fusion of endosomes with lysosomes, and retrograde traffic to the Golgi complex. Despite this global inhibition of vesicular fusion, which was accompanied by gross alterations in cell morphology, SOC activation persisted. These observations cannot be easily reconciled with the vesicle-mediated coupling hypothesis of SOC activation. Our findings imply that the SOC and the machinery necessary to activate them exist in the plasma membrane or are associated with it prior to activation. PMID- 12764155 TI - Hepatitis C virus core protein differently regulates the JAK-STAT signaling pathway under interleukin-6 and interferon-gamma stimuli. AB - We established hepatitis C virus (HCV) core-expressing cells and investigated whether HCV core would modify the Janus kinase (JAK)-signal transducer and activator transcription factor (STAT) pathway under interleukin-6 (IL-6) and interferon (IFN)-gamma stimuli. Phosphorylation of JAK1/2 and STAT3, and STAT3 mediated transcription, were prevented by HCV core under IL-6 stimulation. In contrast, HCV core increased phosphorylation of JAK1/2 and STAT1 and STAT1 mediated transcription under IFN-gamma stimulation. Immunoprecipitation/Western blot analysis showed that HCV core could bind to JAK1/2. The PGYPWP sequences at codons 79-84 within HCV core were important for interaction with JAKs by in vitro binding analysis. In the reporter gene assay, HCV core-mediated suppression of JAK-STAT pathway under IL-6 stimulation was not observed by abrogation of PGYPWP sequence, suggesting that HCV core/JAK interaction may directly affect the signal transduction. In contrast, augmentation of JAK-STAT pathway was still seen by HCV core without functional PGYPWP sequence under IFN-gamma stimulation. Flow cytometric analysis revealed that HCV core up-regulated of IFN-gamma receptor 2 expression, which may be responsible for HCV core-mediated enhancement of JAK STAT pathway under IFN-gamma stimulation. In conclusion, HCV core has different effects on the JAK-STAT pathway under IL-6 and IFN-gamma stimuli. This may be exerted by these two independent mechanisms. PMID- 12764156 TI - Truncation of the A1 adenosine receptor reveals distinct roles of the membrane proximal carboxyl terminus in receptor folding and G protein coupling. AB - The carboxyl terminus (C-tail) of G protein-coupled receptors is divergent in length and structure and may represent an individualized cytoplasmic domain. By progressively truncating the A1 adenosine receptor, a Gi/o-coupled receptor with short cytoplasmic stretches, we identify two inherent functions of the C-tail, namely a role in receptor export from the endoplasmic reticulum (ER) and a role in G protein coupling. Deletion of the last 22 and 26 amino acids (of 36) reduced and completely abolished surface expression of the receptor, respectively. The severely truncated receptors were retained in the ER and failed to bind ligands. If overexpressed, even a substantial portion of the full-length receptor was retained in the ER in a form that was not functional. These data indicate that folding is rate limiting in export from the ER and that the proximal segment of the carboxyl terminus provides a docking site for the machinery involved in folding and quality control. In addition, the proximal portion is also important in G protein coupling. This latter role was unmasked when the distal portion of the C-tail (the extreme 18 amino acids, including a palmitoylated cysteine) had been removed; the resulting receptor was functional and transferred the agonist mediated signal more efficiently than the full-length receptor. Signaling was enhanced because the coupling affinity increased (by 3-fold), which translated into a higher agonist potency. Thus, the distal portion of the carboxyl terminus provides for an autoinhibitory restraint, presumably by folding back and preventing G protein access to the proximal part of the C-tail. PMID- 12764157 TI - Residues in the extracellular loop 4 are critical for maintaining the conformational equilibrium of the gamma-aminobutyric acid transporter-1. AB - We mutated residues Met345 and Thr349 in the rat gamma-aminobutyric acid transporter-1 (GAT-1) to histidines (M345H and T349H). These two residues are located four amino acids apart at the extracellular end of transmembrane segment 7 in a region of GAT-1 that we have previously suggested undergoes conformational changes critical for the transport process. The two single mutants and the double mutant (M345H/T349H) were expressed in Xenopus laevis oocytes, and their steady state and presteady-state kinetics were examined and compared with wild type GAT 1 by using the two-electrode voltage clamp method. Oocytes expressing M345H showed a decrease in apparent GABA affinity, an increase in apparent affinity for Na+, a shift in the charge/voltage (Q/Vm) relationship to more positive membrane potentials, and an increased Li+-induced leak current. Oocytes expressing T349H showed an increase in apparent GABA affinity, a decrease in apparent Na+ affinity, a profound shift in the Q/Vm relationship to more negative potentials, and a decreased Li+-induced leak current. The data are consistent with a shift in the conformational equilibrium of the mutant transporters, with M345H stabilized in an outward-facing conformation and T349H in an inward-facing conformation. These data suggest that the extracellular end of transmembrane domain 7 not only undergoes conformational changes critical for the translocation process but also plays a role in regulating the conformational equilibrium between inward- and outward-facing conformations. PMID- 12764158 TI - Committed to the best science? PMID- 12764159 TI - AIDS research. Drug trials without the drugs? PMID- 12764161 TI - Climate. A surprise La Nina. PMID- 12764160 TI - European Space Agency. Financial crisis puts comet mission on the ropes. PMID- 12764162 TI - Neuroscience. Monarchs check clock to chart migration route. PMID- 12764163 TI - SARS. Researcher told to stay home after China trip. PMID- 12764164 TI - Royal Society. Nine women make 2003 a record year. PMID- 12764165 TI - Solar cells. Tricks for beating the heat help panels see the light. PMID- 12764166 TI - Kazakhstan. Plutonium fields forever. PMID- 12764167 TI - Kazakhstan. For a long-suffering population, uncertainty reigns. PMID- 12764168 TI - Kazakhstan. Safe haven for a breeder's plutonium hoard. PMID- 12764170 TI - European facilities. Stuck in neutron neutral. PMID- 12764169 TI - Clinical trials. In critical condition. PMID- 12764171 TI - Albert Osterhaus profile. The virus collector. PMID- 12764172 TI - Medical Research Council. Out of the frying pan into the MRC. PMID- 12764173 TI - Science, ethics, and war. PMID- 12764175 TI - Carbon nanotubes provide a charge. PMID- 12764174 TI - Supporting scientists and research in Iraq. PMID- 12764176 TI - Inoculating a science education epidemic. PMID- 12764177 TI - Comment on "Single Crystals of Single-Walled Carbon Nanotubes Formed by Self Assembly". PMID- 12764179 TI - Ecology. Three-Gorges Dam--experiment in habitat fragmentation? AB - Habitat fragmentation is the primary cause of the loss of biodiversity and ecosystem services, but its underlying processes and mechanisms remain poorly understood. Studies of islands and insular terrestrial habitats are essential for improving our understanding of habitat fragmentation. We argue that the Three Gorges Dam, the largest that humans have ever created, presents a unique grand scale natural experiment that allows ecologists to address a range of critical questions concerning the theory and practice of biodiversity conservation. PMID- 12764180 TI - Ecology. Protecting China's biodiversity. PMID- 12764181 TI - Atmospheric science. Slow in, rapid out--carbon flux studies and Kyoto targets. PMID- 12764183 TI - Materials science. Nanocrystals get twins. PMID- 12764182 TI - Plant sciences. Super sunflowers--stopping the rot? PMID- 12764184 TI - Applied physics. Transparent electronics. PMID- 12764185 TI - Genomics. Not junk after all. PMID- 12764186 TI - Retrospective. "My little spontaneous blips". PMID- 12764187 TI - Growth of epitaxial nanowires at the junctions of nanowalls. PMID- 12764188 TI - Fitness effects of transgenic disease resistance in sunflowers. PMID- 12764189 TI - Keeping G proteins at bay: a complex between G protein-coupled receptor kinase 2 and Gbetagamma. AB - The phosphorylation of heptahelical receptors by heterotrimeric guanine nucleotide-binding protein (G protein)-coupled receptor kinases (GRKs) is a universal regulatory mechanism that leads to desensitization of G protein signaling and to the activation of alternative signaling pathways. We determined the crystallographic structure of bovine GRK2 in complex with G protein beta1gamma2 subunits. Our results show how the three domains of GRK2-the RGS (regulator of G protein signaling) homology, protein kinase, and pleckstrin homology domains-integrate their respective activities and recruit the enzyme to the cell membrane in an orientation that not only facilitates receptor phosphorylation, but also allows for the simultaneous inhibition of signaling by Galpha and Gbetagamma subunits. PMID- 12764190 TI - Orbital motion in the radio galaxy 3C 66B: evidence for a supermassive black hole binary. AB - Supermassive black hole binaries may exist in the centers of active galactic nuclei such as quasars and radio galaxies, and mergers between galaxies may result in the formation of supermassive binaries during the course of galactic evolution. Using the very-long-baseline interferometer, we imaged the radio galaxy 3C 66B at radio frequencies and found that the unresolved radio core of 3C 66B shows well-defined elliptical motions with a period of 1.05 +/- 0.03 years, which provides a direct detection of a supermassive black hole binary. PMID- 12764191 TI - Ferroelectricity in free niobium clusters. AB - Electric deflections of gas-phase, cryogenically cooled, neutral niobium clusters [NbN; number of atoms (N) = 2 to 150, temperature (T) = 20to 300kelvin], measured in molecular beams, show that cold clusters may attain an anomalous component with very large electric dipole moments. In contrast, room-temperature measurements show normal metallic polarizabilities. Characteristic energies kBTG(N) [Boltzmann constant kB times a transition temperature TG(N)] are identified, below which the ferroelectric-like state develops. Generally, TG decreases [110 > TG(N) > 10K] as N increases, with pronounced even-odd alternations for N > 38. This new state of metallic matter may be related to bulk superconductivity. PMID- 12764192 TI - Thin-film transistor fabricated in single-crystalline transparent oxide semiconductor. AB - We report the fabrication of transparent field-effect transistors using a single crystalline thin-film transparent oxide semiconductor, InGaO3(ZnO)5, as an electron channel and amorphous hafnium oxide as a gate insulator. The device exhibits an on-to-off current ratio of approximately 106 and a field-effect mobility of approximately 80 square centimeters per volt per second at room temperature, with operation insensitive to visible light irradiation. The result provides a step toward the realization of transparent electronics for next generation optoelectronics. PMID- 12764193 TI - Geometric origin of hexagonal close packing at a grain boundary in gold. AB - Using electron microscopy, we identify local, intergranular regions of hexagonal close-packing at a grain boundary in gold. By analyzing the topological defects that connect this layer to the adjacent face-centered cubic grains, we explain the geometric origin of this interfacial reconstruction. We extend this analysis to predict the stacking arrangements found over a range of intergranular misorientations. These results help to unify our understanding of the defects that control the behavior of polycrystalline materials by showing how line defects that are already well understood in the bulk also can determine the atomic arrangements at grain boundaries. PMID- 12764194 TI - n-Type conducting CdSe nanocrystal solids. AB - A bottleneck limiting the widespread application of semiconductor nanocrystal solids is their poor conductivity. We report that the conductivity of thin films of n-type CdSe nanocrystals increases by many orders of magnitude as the occupation of the first two electronic shells, 1Se and 1Pe, increases, either by potassium or electrochemical doping. Around half-filling of the 1Se shell, a peak in the conductivity is observed, indicating shell-to-shell transport. Introducing conjugated ligands between nanocrystals increases the conductivities of these states to approximately 10(-2) siemens per centimeter. PMID- 12764195 TI - A modular PIP2 binding site as a determinant of capsaicin receptor sensitivity. AB - The capsaicin receptor (TRPV1), a heat-activated ion channel of the pain pathway, is sensitized by phosphatidylinositol-4,5-bisphosphate (PIP2) hydrolysis after phospholipase C activation. We identify a site within the C-terminal domain of TRPV1 that is required for PIP2-mediated inhibition of channel gating. Mutations that weaken PIP2-TRPV1 interaction reduce thresholds for chemical or thermal stimuli, whereas TRPV1 channels in which this region is replaced with a lipid binding domain from PIP2-activated potassium channels remain inhibited by PIP2. The PIP2-interaction domain therefore serves as a critical determinant of thermal threshold and dynamic sensitivity range, tuning TRPV1, and thus the sensory neuron, to appropriately detect heat under normal or pathophysiological conditions. PMID- 12764196 TI - The birth of an alternatively spliced exon: 3' splice-site selection in Alu exons. AB - Alu repetitive elements can be inserted into mature messenger RNAs via a splicing mediated process termed exonization. To understand the molecular basis and the regulation of the process of turning intronic Alus into new exons, we compiled and analyzed a data set of human exonized Alus. We revealed a mechanism that governs 3' splice-site selection in these exons during alternative splicing. On the basis of these findings, we identified mutations that activated the exonization of a silent intronic Alu. PMID- 12764198 TI - CD2-associated protein haploinsufficiency is linked to glomerular disease susceptibility. AB - Loss of CD2-associated protein (CD2AP), a component of the filtration complex in the kidney, causes death in mice at 6 weeks of age. Mice with CD2AP haploinsufficiency developed glomerular changes at 9 months of age and had increased susceptibility to glomerular injury by nephrotoxic antibodies or immune complexes. Electron microscopic analysis of podocytes revealed defects in the formation of multivesicular bodies, suggesting an impairment of the intracellular degradation pathway. Two human patients with focal segmental glomerulosclerosis had a mutation predicted to ablate expression of one CD2AP allele, implicating CD2AP as a determinant of human susceptibility to glomerular disease. PMID- 12764199 TI - Uncoupling of leading- and lagging-strand DNA replication during lesion bypass in vivo. AB - Numerous agents attack DNA, forming lesions that impair normal replication. Specialized DNA polymerases transiently replace the replicative polymerase and copy past lesions, thus generating mutations, the major initiating cause of cancer. We monitored, in Escherichia coli, the kinetics of replication of both strands of DNA molecules containing a single replication block in either the leading or lagging strand. Despite a block in the leading strand, lagging-strand synthesis proceeded further, implying transient uncoupling of concurrent strand synthesis. Replication through the lesion requires specialized DNA polymerases and is achieved with similar kinetics and efficiencies in both strands. PMID- 12764197 TI - Essential role of Fkbp6 in male fertility and homologous chromosome pairing in meiosis. AB - Meiosis is a critical stage of gametogenesis in which alignment and synapsis of chromosomal pairs occur, allowing for the recombination of maternal and paternal genomes. Here we show that FK506 binding protein (Fkbp6) localizes to meiotic chromosome cores and regions of homologous chromosome synapsis. Targeted inactivation of Fkbp6 in mice results in aspermic males and the absence of normal pachytene spermatocytes. Moreover, we identified the deletion of Fkbp6 exon 8 as the causative mutation in spontaneously male sterile as/as mutant rats. Loss of Fkbp6 results in abnormal pairing and misalignments between homologous chromosomes, nonhomologous partner switches, and autosynapsis of X chromosome cores in meiotic spermatocytes. Fertility and meiosis are normal in Fkbp6 mutant females. Thus, Fkbp6 is a component of the synaptonemal complex essential for sex specific fertility and for the fidelity of homologous chromosome pairing in meiosis. PMID- 12764200 TI - Illuminating the circadian clock in monarch butterfly migration. AB - Migratory monarch butterflies use a time-compensated Sun compass to navigate to their overwintering grounds in Mexico. Here, we report that constant light, which disrupts circadian clock function at both the behavioral and molecular levels in monarchs, also disrupts the time-compensated component of flight navigation. We further show that ultraviolet light is important for flight navigation but is not required for photic entrainment of circadian rhythms. Tracing these distinct light-input pathways into the brain should aid our understanding of the clock compass mechanisms necessary for successful migration. PMID- 12764201 TI - Europe's terrestrial biosphere absorbs 7 to 12% of European anthropogenic CO2 emissions. AB - Most inverse atmospheric models report considerable uptake of carbon dioxide in Europe's terrestrial biosphere. In contrast, carbon stocks in terrestrial ecosystems increase at a much smaller rate, with carbon gains in forests and grassland soils almost being offset by carbon losses from cropland and peat soils. Accounting for non-carbon dioxide carbon transfers that are not detected by the atmospheric models and for carbon dioxide fluxes bypassing the ecosystem carbon stocks considerably reduces the gap between the small carbon-stock changes and the larger carbon dioxide uptake estimated by atmospheric models. The remaining difference could be because of missing components in the stock-change approach, as well as the large uncertainty in both methods. With the use of the corrected atmosphere- and land-based estimates as a dual constraint, we estimate a net carbon sink between 135 and 205 teragrams per year in Europe's terrestrial biosphere, the equivalent of 7 to 12% of the 1995 anthropogenic carbon emissions. PMID- 12764202 TI - Atomic memory for correlated photon states. AB - We experimentally demonstrate emission of two quantum-mechanically correlated light pulses with a time delay that is coherently controlled via temporal storage of photonic states in an ensemble of rubidium atoms. The experiment is based on Raman scattering, which produces correlated pairs of spin-flipped atoms and photons, followed by coherent conversion of the atomic states into a different photon beam after a controllable delay. This resonant nonlinear optical process is a promising technique for potential applications in quantum communication. PMID- 12764204 TI - Media warnings about environmental pollution facilitate the acquisition of symptoms in response to chemical substances. AB - OBJECTIVE: Previous studies showed that somatic symptoms can be acquired in response to chemical substances using an associative learning paradigm, but only when the substance was foul smelling and not when it smelled pleasant. In this study, we investigated whether warnings about environmental pollution would facilitate acquiring symptoms, regardless of the pleasantness of the smell. METHOD: One group received prior information framing the study in the context of the rapidly increasing chemical pollution of our environment. Another group received no prior information. Conditional odor stimuli (CS) were diluted ammonia (foul-smelling) and niaouli (neutral-positive smelling); the unconditional stimulus (UCS) was 10% CO2-enriched air. Each subject breathed one odor mixed with CO2 and a control odor mixed with air in 80-sec breathing trials. The type of odor mixed with CO2 was counterbalanced across participants. Next, the same breathing trials were administered without CO2. Breathing behavior was measured during each trial; subjective symptoms were assessed after each trial. RESULTS: Only participants who had been given warnings about environmental pollution reported more symptoms to the odor that had previously been associated with CO2, compared with the control odor. This was so for both the foul- and the pleasant smelling odor. Symptom learning did not occur in the group that did not receive warnings. The elevated symptom level could not be accounted for by altered respiratory behavior, nor by experimental demand effects. CONCLUSIONS: Raising environmental awareness through warnings about chemical pollution facilitates learning of subjective health symptoms in response to chemical substances. PMID- 12764205 TI - Is type A behavior really a trigger for coronary heart disease events? AB - OBJECTIVE: The purpose of this study was to compare chronic with acute mechanisms by which Type A might predict incident coronary heart disease (CHD). METHOD: The study included 2394 men aged 50 to 64 years who were assessed for CHD, Type A behavior, and CHD risk factors. Type A was assessed using the Jenkins Activity Survey (JAS), the Bortner scale, and the Framingham scale. Further examinations were completed at 5 and 9 years for incident CHD. RESULTS: After 9 years, there was no increased risk of CHD associated with any Type A score. Nevertheless, high Bortner scores were associated with increased risk of incident CHD at 5 years and high JAS and Bortner scores were associated with a decreased risk between 5 and 9 years. Further analysis of Type A scores on time to first coronary event found strong inverse associations for all type A scores (JAS = 205 -0.49 months to first event, 95% CI = -0.20, -0.78, p =.001) (Bortner = 176 -0.27 months; 95% CI = -0.10, -0.44; p =.002) (Framingham = 0.44 -0.0011 months; 95% CI = -0.0002, 0.0019; p =.01). CONCLUSIONS: The data show Type A is a strong predictor of when incident coronary heart disease (or coronary event) will occur rather than if it will occur. These findings suggest that Type A increases exposure to potential triggers, rather than materially affecting the process of atherosclerosis. PMID- 12764207 TI - Postdelivery screening for postpartum depression. AB - OBJECTIVE: Postpartum depression (PPD) is a serious psychiatric disorder affecting 10% to 20% of women after childbirth. Research has shown that systematic screening for PPD using self-report questionnaires helps improve the identification of PPD and expedite treatment. Most studies on PPD screening have been conducted in the second and third postpartum months; little is known about whether PPD screening can be carried out on the days immediately after delivery. METHODS: A prospective cohort of 145 women completed the Edinburgh Postnatal Depression Scale (EPDS), Beck Depression Inventory (BDI), and General Health Questionnaire (GHQ) within 2 days of delivery. Six weeks after delivery, the participants were interviewed by a psychiatrist, who used the Structured Clinical Interview for DSM-III-R (SCID, nonpatient version) to establish the diagnosis. The psychometric performance of the EPDS, BDI, and GHQ in detecting PPD was assessed using the SCID diagnosis as the gold standard. RESULTS: When the cutoffs of the EPDS, BDI, and GHQ were lowered to achieve a sensitivity of 80%, the positive predictive values of these scales were to 13%, 18%, and 21%, respectively. When the cutoffs were raised to achieve a positive predictive value of 50%, the sensitivity rates were 6% (EPDS), 14% (GHQ), and 36% (BDI). CONCLUSIONS: When commonly used depression rating scales were administered to identify PPD immediately after delivery, their psychometric properties were unsatisfactory. Healthcare providers should not screen for PPD in the first few days after delivery. PMID- 12764206 TI - Association between depression and elevated C-reactive protein. AB - OBJECTIVE: Depression has been related to a higher risk of developing coronary heart disease, but the mechanism that accounts for this association is unclear. Because atherosclerosis is an inflammatory process, depression could increase the risk of coronary heart disease by inducing or promoting inflammation. The objective of the present study was to investigate the association between history of major depressive episode and presence of low-grade systemic inflammation as measured by serum C-reactive protein (CRP). METHODS: We analyzed data from the Third National Health and Nutrition Examination Survey, a representative sample of the US population from 1988 to 1994. Participants included a total of 6149 individuals aged 17 to 39 years who were free of cardiovascular diseases and chronic inflammatory conditions. The main predictor variable of interest was lifetime history of a major depressive episode as assessed by means of the Diagnostic Interview Schedule. The main outcome variable was the presence or absence of an elevated CRP level (> or =22 mg/dl). RESULTS: Among men, history of a major depressive episode was associated with elevated CRP, particularly for recent episodes (up to 6 months before assessment). In multivariate analyses, men with a history of major depressive episode had 2.77 times higher odds of elevated CRP compared with never-depressed men (95% confidence interval, 1.43-5.26). The adjusted odds ratio was 3.81, 3.98, 1.51, and 1.52 for men who had their last major depressive episode less than 1 month before, 1 to 6 months before, 7 to 12 months before, and more than 12 months before assessment, respectively (p for trend =.004). In women, a comparable association between depression and CRP was quite weak and not significant. CONCLUSIONS: A recent history of major depressive episode is strongly associated with elevated CRP in men aged 17 to 39. In this group, low-grade systemic inflammation could represent a mechanism linking depression to cardiovascular risk. PMID- 12764208 TI - The relation of severity of depressive symptoms to monocyte-associated proinflammatory cytokines and chemokines in apparently healthy men. AB - OBJECTIVE: We examined the relation of severity of depressive symptoms to lipopolysaccharide-stimulated expression of monocyte-associated proinflammatory cytokines and chemokines in 53 nonsmoking, healthy men. METHODS: Assessments of cytokine and chemokine expression and severity of depressive symptoms were conducted on the same day. The 21-item Beck Depression Inventory (BDI) was used to assess severity of depressive symptoms experienced during the week before study participation. Dual-color flow cytometry was used to determine monocyte associated (CD14+) expression of interleukin-1alpha (IL-1alpha), IL-1beta, tumor necrosis factor-alpha (TNF-alpha), IL-8, and monocyte chemotactic protein-1 (MCP 1) after in vitro lipopolysaccharide stimulation of undiluted whole blood. RESULTS: Calculations of partial correlation coefficients controlling for age, race, body mass index, and alcohol use indicated that BDI score was significantly associated with IL-1alpha (r = 0.27), IL-1beta (r = 0.44), TNF-alpha (r = 0.57), MCP-1 (r = 0.52), and IL-8 (r = 0.33). In addition, relative to men with BDI scores below 10, men with BDI scores of 10 or above exhibited an overexpression of IL-1beta (p =.004), TNF-alpha (p =.005), IL-8 (p =.002), and MCP-1 (p =.025). CONCLUSIONS: Relative to men with no or minimal symptoms of depression, men with mild to moderate levels of depressive symptoms showed overexpression of monocyte associated proinflammatory cytokines and chemokines. PMID- 12764209 TI - Impact of depression on experimental pain perception: a systematic review of the literature with meta-analysis. AB - OBJECTIVE: This systematic review and meta-analysis was performed to examine the impact of depression on the perception of experimental pain stimuli. METHODS: CD ROM databases and bibliographies were searched to identify studies comparing the psychophysical responses to experimental pain stimuli of depressed subjects with that of healthy controls. Effect sizes (Cohen's d) and probabilities were combined across studies; positive effect sizes indicated higher thresholds in depressed groups. RESULTS: Six methodologically rigorous, independent studies were found comparing psychophysical responses to experimental pain stimuli in depressed subjects and healthy controls. Pain perception threshold was higher in depressed subjects (6 studies, d = 0.38, p =.001). This finding was not the result of publication bias. Absolute sensory perception threshold was much higher in depressed subjects (2 studies, d = 0.68, p =.002), though the findings for pain tolerance (2 studies) were too heterogeneous to enable us to combine results. CONCLUSIONS: Depressed subjects are less likely to perceive a sensory stimulus as being painful compared with nondepressed controls. The influence of depression on attention to the pain stimulus may account for this effect. More studies are required to enable us to determine the impact of depression on absolute sensory perception threshold and pain tolerance. Furthermore, more studies would enable the examination of depression on the perception of different modalities. PMID- 12764210 TI - Depression and coronary heart disease in women with diabetes. AB - OBJECTIVE: The protective effects of female gender on the appearance and course of coronary heart disease (CHD) in nondiabetic subjects are diminished in the presence of diabetes. Depression predicts onset of and poor outcome from CHD in nondiabetic populations. We hypothesized that the doubled rates of depression in female diabetic patients could help explain the high prevalence of CHD in women with diabetes. METHOD: Seventy-six female type 1 and type 2 diabetic patients with (N=16) or without (N=60) active major depression (DSM-III) at index evaluation underwent systematic annual investigation of diabetes and its complications for up to 10 years. Occurrences of CHD and other macrovascular complications were examined in relation to depression status using survival analysis statistics. A multivariate model incorporating other CHD risk factors (age, duration of diabetes, body mass index, glycosylated hemoglobin, and presence of hypertension, hyperlipidemia, or tobacco use) was used to determine independent effects of depression on outcome. RESULTS: Development of CHD was significantly more rapid in the depressed subset (p<0.01 between 10-year curves), an effect that persisted after controlling for base-line differences in body mass index. Depression also was retained as an independent predictor of CHD in the multivariate model with an age-adjusted hazard ratio of 5.2 (95% CI: 1.4-18.9; p=.01). In contrast, depression did not predict the development of clinically apparent peripheral or cerebrovascular disease. CONCLUSIONS: In this sample of diabetic women, major depression was an independent risk factor that accelerated the development of CHD. Depression recognition and management may improve outcomes from diabetes in this gender subgroup. PMID- 12764211 TI - Autoregulation of blood pressure and thought: preliminary results of an application of brain imaging to psychosomatic medicine. AB - OBJECTIVE: This presentation seeks to demonstrate the use of brain imaging techniques for understanding the interaction between hypertension and psychosocial function. METHODS: The historical background for the study of brain function among hypertensive patients is reviewed. An initial and a current project examining rCBF with 15O water radiotracer and PET in unmedicated hypertensives and normotensives are described. The rCBF response is assessed during the performance of spatial and verbal working memory tasks of increasing memory load. The assessment also addresses the influence on rCBF and performance of white matter hyperintensities and the presence of carotid artery thickening. RESULTS: Initial results suggest that hypertensives relative to normotensives show less CBF and less posterior parietal rCBF in response to increases in memory load. Hypertensives, however, increase lateral prefrontal (Broca's area)/insula and amygdala/hippocampal rCBF more than normotensives. CONCLUSION: Initial results are sufficient to show that hypertension induces changes in rCBF. A tentative hypothesis is that a relatively general decrease in rCBF responsivity induces specific compensatory cognitive strategies as well as subcortical activation. The rCBF changes appear to have implications for information processing and, as such, hold promise for understanding prior reports relating hypertension to affective regulation and cardiovascular reactivity. Imaging techniques provide a powerful tool for psychosomatic research. PMID- 12764212 TI - Risk factors for myocardial infarction during vacation travel. AB - OBJECTIVES: Medical emergencies occur increasingly outside the usual health care area as a result of increased leisure and professional travel. Acute coronary syndromes are the leading cause of mortality during vacation. Vacation activities include physical and emotional triggers for myocardial infarction (MI). This study examines characteristics of vacation travel as risk factors for MI. METHODS: Patients diagnosed with MI during vacation abroad (N = 92; age, 59.5 +/- 10.2; 79 men) were recruited through an emergency health insurance organization. Risk indicators for Vacation MI were examined and included: cardiovascular risk factors, psychosocial measures, and specific demands and activities related to vacation (eg, lodging accommodations, unfamiliar destination, mode of transportation, short-term planning). Vacation MI patients were compared with two reference groups: age-matched Vacation Controls with noncardiovascular medical emergencies (N = 67) and Hospital MI Controls, admitted in their usual health care area (N = 30). RESULTS: Vacation MI occurred disproportionately (21.1%) during the first 2 days of vacation. Cardiovascular risk factors were more prevalent among Vacation MI patients than Vacation Controls (p values <.05) but not compared with Hospital MI Controls. Vacation MI occurred more often in patients with lower education (OR = 2.4, CI = 1.1-5.2) and those living with a spouse (OR = 2.6, CI = 1.0-7.1) than age-matched Vacation Controls. Compared with Hospital MI Controls, Vacation MI occurred more often among patients traveling by car versus other modes of transportation (OR = 2.5, CI = 1.0-6.1) and among patients staying in a tent or mobile home versus hotel (OR = 9.7, CI = 2.0-47.9). CONCLUSION: Incidence of MI during vacation is highest during the first 2 days of vacation. Vacation activities such as adverse driving conditions and less luxurious accommodations may increase risk for MI. Individuals with known vulnerability for MI may therefore benefit from minimizing physical and emotional challenges specifically related to vacation travel. PMID- 12764213 TI - Associations between psychological traits and endothelial function in postmenopausal women. AB - OBJECTIVE: The objective of this study was to determine whether psychosocial risk factors for cardiovascular disease (CVD) are associated with impairments in endothelial function and to determine whether use of hormone replacement therapy (HRT) can moderate observed associations among women without prior CVD. METHODS: Flow-mediated dilation was assessed by brachial ultrasound after reactive hyperemia in 193 postmenopausal women enrolled in the prospective Healthy Women Study. Measures of psychosocial characteristics had been completed at study entry, when the women were premenopausal (mean = 13.6 years earlier), and at a separate postmenopausal follow-up exam near the time of the ultrasound (mean = 1.5 years earlier). RESULTS: Factor analyses of the psychosocial characteristics yielded two factors: Type A/anger and anxiety/depression. Anxiety/depression scores at the study entry and follow-up exams and Type A/anger scores at the study entry exam were associated with less vasodilation (p values < 0.05). Type A/anger scores at the follow-up exam were associated with less vasodilation among women not using HRT (p <.05). CONCLUSIONS: Psychosocial risk factors for CVD are associated with impaired brachial artery dilation among postmenopausal women. HRT use may mask some associations between psychosocial risk factors and endothelial dysfunction among postmenopausal women. PMID- 12764214 TI - Cardiovascular reactivity to stress predicts future blood pressure in adolescence. AB - OBJECTIVE: This study evaluated the prospective relationship between cardiovascular reactivity to psychological stress and increases in resting blood pressure across a 3-year period among a multiethnic pediatric sample (N = 149). METHODS: Systolic and diastolic blood pressure; EKG heart rate, pre-ejection period, and mean successive difference of R to R intervals; and impedance-derived measures of cardiac output, stroke volume, and total peripheral resistance were collected during performance of four tasks that elicited different hemodynamic response patterns. Changes from baseline to each task were standardized and averaged to form eight composite scores. Analyses adjusted for time 1 baseline blood pressure and age, body mass index at baseline and change to follow-up, and duration of follow-up. RESULTS: Rises in SBP over the follow-up period were independently predicted by SBP (beta = 0.161, p =.009), DBP (beta = 0.132, p =.02), and CO (beta = 0.144, p =.02) composite measures of reactivity. Rises in DBP over the follow-up period were predicted by DBP (beta = 0.292, p =.003, respectively), and MSD (beta = -0.176, p <.03) composite measures of reactivity. TPR reactivity was not related prospectively to blood pressure rises. CONCLUSIONS: This study adds to the pediatric literature documenting an association between cardiovascular reactivity to stress and subsequent risk for hypertension. It is the first to show that impedance-derived measures of myocardial function during stress are related to future blood pressure levels. PMID- 12764215 TI - Psychological distress and pain significantly increase before death in metastatic breast cancer patients. AB - OBJECTIVE: This study was designed to examine the course of psychological distress and pain from study entry to death in 59 women with metastatic breast cancer participating in a randomized trial of the effects of group psychotherapy on psychosocial outcomes and survival. It was hypothesized that psychological distress would increase significantly before death independent of changes in pain. METHOD: Data were collected as part of a larger study (N = 125). Analyses were based on data from a subset of women who had died and for whom we had data from at least three assessments. Mean levels of mood, trauma symptoms, depression symptoms, well-being, and pain over three time points were examined: at baseline (T1), the second-to-last assessment before death (T2), and the last assessment before death (T3). RESULTS: Results indicate that while psychological distress remained relatively constant or declined from T1 to T2, means on all measures significantly changed in the hypothesized direction from T2 to T3. Neither self reported pain, nor the passage of time, appeared to account for these changes. Additionally, participation in group psychotherapy did not have a significant impact on this change in distress proximal to death. CONCLUSIONS: Results suggest that specialized end-stage clinical interventions are particularly needed for cancer patients as they approach death. Moreover, intervention studies for patients with deteriorating illnesses may need to take this "spike" in psychological distress and pain proximal to death into account to avoid Type II errors in evaluations of psychological outcomes. PMID- 12764216 TI - Life stress and cervical squamous intraepithelial lesions in women with human papillomavirus and human immunodeficiency virus. AB - OBJECTIVE: Human immunodeficiency virus (HIV)-infected women are at risk for cervical intraepithelial neoplasia (CIN) and cancer due to impaired immunosurveillance over human papillomavirus (HPV) infection. Life stress has been implicated in immune decrements in HIV-infected individuals and therefore may contribute to CIN progression over time. The purpose of this study was to determine whether life stress was associated with progression and/or persistence of squamous intraepithelial lesions (SIL), the cytologic diagnosis conferred by Papanicolaou smear, after 1-year follow-up among women co-infected with HIV and HPV. METHOD: Thirty-two HIV-infected African-American and Caribbean-American women underwent a psychosocial interview, blood draw, colposcopy, and HPV cervical swab at study entry. Using medical chart review, we then abstracted SIL diagnoses at study entry and after 1-year follow-up. RESULTS: Hierarchical logistic regression analysis revealed that higher life stress increased the odds of developing progressive/persistent SIL over 1 year by approximately seven-fold after covarying relevant biological and behavioral control variables. CONCLUSIONS: These findings suggest that life stress may constitute an independent risk factor for SIL progression and/or persistence in HIV-infected women. Stress management interventions may decrease risk for SIL progression/persistence in women living with HIV. PMID- 12764217 TI - Coping and distress: what happens after intervention? A 2-year follow-up from the Swedish Obese Subjects (SOS) study. AB - OBJECTIVE: The study examined effects of weight change on coping and distress in severely obese subjects treated conventionally or undergoing weight reduction surgery. METHODS: We used the Obesity Coping (OC) scale measuring emotion focused, maladaptive coping (Wishful Thinking) and problem-focused, adaptive coping (Social Trust and Fighting Spirit). We also used the Obesity Distress (OD) scale (Intrusion and Helplessness) and the Hospital Anxiety and Depression (HAD) scale. A total of 1146 surgical candidates and 1085 conventionally treated patients completed the OC and OD before treatment and after 24 months. RESULTS: Weight gainers reduced their use of both problem- and emotion-focused coping, thus leaving distress levels unchanged. All participants who lost weight decreased in emotion-focused coping and distress. Participants losing 20 kg or more also increased in problem-focused coping, resulting in even greater improvements regarding distress. CONCLUSIONS: Two years after starting treatment, the pattern and magnitude of change in coping and distress was the same irrespective of type of treatment and was, instead, related to the amount of weight change (the more weight change the greater the changes in coping and distress). Increases in problem-focused coping required major weight reduction, whereas minor weight gain led to a decrease. Emotion-focused coping decreased irrespective of direction of weight change, suggesting a general intervention effect of receiving professional help and support. These results have implications concerning behavior-based interventions of obese patients. PMID- 12764218 TI - Psychosocial factors in patients with noncardiac chest pain. AB - OBJECTIVE: This study sought to explore some psychosocial factors that distinguished individuals with noncardiac chest pain (NCCP) from those without NCCP, and whether these psychosocial factors were associated with anxiety and depression that are co-morbid factors of NCCP. METHODS: A matched case-control design was adopted to compare differences in psychosocial factors among a target group of patients with NCCP (N = 70), a pain control group of patients with rheumatism (N = 70), and a community control group of healthy individuals (N = 70). RESULTS: Compared with subjects from the two control groups, NCCP patients tended to monitor more, use more problem-focused coping, display a coping pattern with a poorer strategy-situation fit, and receive less emotional support in times of stress. Moreover, monitoring perceptual style and problem-focused coping were associated with higher levels of anxiety and depression. Coping pattern with a strategy-situation fit and emotional support were related to lower levels of anxiety and depression. CONCLUSIONS: The present new findings suggest that monitoring perceptual style and inflexible coping style are risk factors that enhance one's vulnerability to NCCP. Emotional support may be a resource factor that reduces one's susceptibility to NCCP. PMID- 12764219 TI - Dissociation between reactivity of the hypothalamus-pituitary-adrenal axis and the sympathetic-adrenal-medullary system to repeated psychosocial stress. AB - OBJECTIVE: This study investigated endocrine and autonomic stress responses after repeated psychosocial stress. A first goal of the study was to investigate whether peripheral catecholamines and cardiovascular parameters would show similar or different habituation patterns after repeated stress. The second aim was to detect possible subgroups with regard to individual habituation patterns in the hypothalamus-pituitary-adrenal (HPA) axis and monitor their respective sympathetic stress responses. METHODS: Sixty-five healthy subjects (19-45 years), 38 men and 27 women, were exposed to the Trier Social Stress Test (TSST) three times with a 4-week interval between stress sessions. Adrenocorticotropic hormone (ACTH), plasma cortisol, salivary cortisol, epinephrine, norepinephrine, and heart rates were measures repeatedly before and after each stress exposure. RESULTS: All endocrine measures as well as heart rates increased significantly after each of the three stress sessions (F values >16.00, all p values <.01). Although salivary free cortisol, total plasma cortisol, ACTH, and heart rate stress responses showed a significant decrease across the three stress sessions (all F values > 5.8, p <.01), no such decrease could be observed for the levels of norepinephrine and epinephrine. A cluster analysis performed on the salivary free cortisol responses to all three stress sessions revealed two response groups consisting of 30 so-called "high responders" and 35 "low responders." The high responders also showed larger ACTH and total plasma cortisol responses compared with the low responders (all F values > 10.00, p <.01). No such differences between high and low responders could be observed with regard to catecholamine and heart rate responses. CONCLUSIONS: From these data we conclude that habituation to psychosocial stress seems to be specific for a given response system. Although HPA responses quickly habituate, the sympathetic nervous system shows rather uniform activation patterns with repeated exposure to psychosocial challenge. PMID- 12764220 TI - Socioeconomic status and stress-related biological responses over the working day. AB - OBJECTIVES: The influence of low socioeconomic status on cardiovascular disease may be mediated in part by sustained activation of stress-related autonomic and neuroendocrine processes. We hypothesized that low socioeconomic status would be associated with heightened ambulatory blood pressure and cortisol output over the working day. METHODS: One hundred eight men and 94 women from the Whitehall II epidemiological cohort participated. Blood pressure and heart rate were monitored every 20 minutes over a working day and evening, and salivary cortisol was sampled on waking up and at 2-hour intervals. Measures were also taken under resting laboratory conditions. Socioeconomic status was indexed by grade of employment. RESULTS: Resting blood pressure, heart rate, and cortisol did not differ by grade. Ambulatory systolic pressure was greater in the morning in the lower (128.9 +/- 15.7 mm Hg) than the intermediate (122.6 +/- 12.5 mm Hg) and higher grades (123.3 +/- 12.7 mm Hg) after adjustment for age, sex, smoking, and alcohol intake (p =.019). Heart rate was also raised in the morning in the lower grade participants. Differences in morning systolic pressure and heart rate were independent of concurrent physical activity. Cortisol concentration was greater in lower than higher grade men (9.54 +/- 4.1 vs. 7.38 +/- 2.8 nmol/liter, p =.008) but was more elevated in higher than lower grade women (7.84 +/- 2.5 vs. 6.35 +/- 1.9 nmol/liter, p =.014). Differences remained significant after adjustment for age, time of awakening, smoking, and alcohol intake. CONCLUSIONS: Socioeconomic differences in blood pressure and cortisol may reflect stress related activation of biological pathways that contribute to variations in disease risk. PMID- 12764221 TI - Association between the type 4 dopamine receptor gene polymorphism and novelty seeking. AB - OBJECTIVE: Mixed results have been reported on the association between the type 4 dopamine receptor gene (DRD4) and the temperament dimension of novelty seeking. We tested this association by specifying the analysis to components of novelty seeking. METHODS: Participants were 150 high and low novelty-seeking scorers (the highest and lowest 10%) from a randomized, population-based sample of Finnish citizens in six age cohorts. We genotyped a 48-bp repeat polymorphism in the DRD4 gene. Novelty seeking was assessed by the Temperament and Character Inventory. RESULTS: No difference in overall novelty seeking between individuals with no seven-repeat allele (short) and any seven-repeat allele (long), between the 4,4 and 4,7 genotype groups, and between long (l/l and s/l) and short (s/s) polymorphism groups were found. The odds ratio for high overall novelty seeking in the presence of any two- or five-repeated alleles vs. none was 2.41 (95% CI, 1.11-5.20). Corresponding odds ratios were significant for exploratory excitability (2.94; 95% CI, 1.32-6.59) and impulsiveness (2.74; 95% CI, 1.23 6.11) but not for other components of novelty seeking. No interactions with age or gender were detected. CONCLUSIONS: The present study confirmed previous findings on the association between the type 4 dopamine receptor gene and novelty seeking, in particular exploratory excitability and impulsiveness. The tendency to avoid or approach a novel situation is a core concept of several temperamental theories. The present findings support the hypothesis that this tendency is associated with DRD4 and might concern temperament psychology in general, not only the concept of novelty seeking. PMID- 12764222 TI - Postoperative course after papilloma resection: effects of written disclosure of the experience in subjects with different alexithymia levels. AB - OBJECTIVE: The aim of the investigation was to assess the effects on postoperative course after bladder papilloma resection of a technique for the written disclosure of traumatic events in interaction with individual differences in alexithymia. METHODS: Forty subjects were administered a general questionnaire and the Toronto Alexithymia Scale (TAS-20) the second day after admittance. Twenty subjects were asked to write for 3 days, 20 minutes a day, about their experience of being in the hospital, following instructions developed by J. W. Pennebaker and coworkers. The postoperative course was assessed objectively by the duration of stay in hospital and subjectively by subjects completing the Symptom Check List 90 (SCL-90) the day before leaving the hospital. RESULTS: Subjects who wrote stayed fewer days in hospital and had lower SCL-90 scores. The same effect was shown by low alexithymia levels. Study of interactions showed that the effect of writing was apparent only in subjects high in alexithymia, whereas subjects low in alexithymia showed a favorable course independent of writing. CONCLUSIONS: Writing about one's thoughts and feelings about being in hospital for a surgical operation has beneficial effects on postoperative course. This holds particularly true for high alexithymic subjects, who obtain through writing the same outcome as low alexithymic subjects. PMID- 12764223 TI - The mediating effects of sleep in the relationship between traumatic stress and health symptoms in urban police officers. AB - OBJECTIVE: Posttraumatic stress symptoms have been associated with increased health problems across numerous studies. Sleep disruption, one of the principal symptoms resulting from traumatic stress, has also been shown to produce health problems. This study explored the hypothesis that the relationship between posttraumatic stress symptoms and health is mediated by sleep problems. METHOD: A sample of 741 police officers were administered measures of traumatic stress symptoms, sleep, health functioning, and somatic symptoms. RESULTS: Traumatic stress symptoms were significantly related to both somatic symptoms (R2 = 0.18, p <.001) and health functioning (R2 = 0.02, p <.01). The relationship between somatic symptoms and traumatic stress symptoms was partially mediated by sleep (p <.001). The relationship between traumatic stress symptoms and health functioning was fully mediated by sleep. CONCLUSIONS: Although design characteristics, such as cross-sectional sampling, limit the inferences that can be drawn, these findings suggest that sleep may serve as an important mediator between traumatic stress and somatic symptoms. PMID- 12764224 TI - Depressive symptoms and metabolic risk in adult male twins enrolled in the National Heart, Lung, and Blood Institute twin study. AB - OBJECTIVE: To determine the extent to which depressive symptoms are associated with metabolic risk factors and whether genetic or environmental factors account for this association. METHOD: Twin structural equation modeling was employed to estimate genetic and environmental contributions to the covariation of depressive symptoms, as indexed by the Centers for Epidemiological Studies-Depression Scale, and common variance among blood pressure, body mass index, waist-to-hip ratio, and serum triglycerides and glucose among 87 monozygotic and 86 dizygotic male twin pairs who participated in the NHLBI twin study. RESULTS: Depressive symptoms were associated with individual components of the metabolic syndrome and common variance among the risk factors. Twin structural equation modeling indicated that the associations were attributable to environmental (nongenetic) factors. CONCLUSIONS: These results support the hypothesis that depressive symptoms may increase risk for a pattern of physiological risk consistent with the metabolic syndrome. PMID- 12764226 TI - Pure-tone birdsong by resonance filtering of harmonic overtones. AB - Pure-tone song is a common and widespread phenomenon in birds. The mechanistic origin of this type of phonation has been the subject of long-standing discussion. Currently, there are three hypotheses. (i) A vibrating valve in the avian vocal organ, the syrinx, generates a multifrequency harmonic source sound, which is filtered to a pure tone by a vocal tract filter ("source-filter" model, analogous to human speech production). (ii) Vocal tract resonances couple with a vibrating valve source, suppressing the normal production of harmonic overtones at this source ("soprano" model, analogous to human soprano singing). (iii) Pure tone sound is produced as such by a sound-generating mechanism that is fundamentally different from a vibrating valve. Here we present direct evidence of a source-filter mechanism in the production of pure-tone birdsong. Using tracheal thermistors and air sac pressure cannulae, we recorded sound signals close to the syringeal sound source during spontaneous, pure-tone vocalizations of two species of turtledove. The results show that pure-tone dove vocalizations originate through filtering of a multifrequency harmonic sound source. PMID- 12764225 TI - Defective importin beta recognition and nuclear import of the sex-determining factor SRY are associated with XY sex-reversing mutations. AB - The architectural transcription factor SRY (sex-determining region of the Y chromosome) plays a key role in sex determination as indicated by the fact that mutations in SRY are responsible for XY gonadal dysgenesis in humans. Although many SRY mutations reduce DNA-binding/bending activity, it is not clear how SRY mutations that do not affect interaction with DNA contribute to disease. The SRY high-mobility group domain harbors two nuclear localization signals (NLSs), and here we examine SRY from four XY females with missense mutations in these signals. In all cases, mutant SRY protein is partly localized to the cytoplasm, whereas wild-type SRY is strictly nuclear. Each NLS can independently direct nuclear transport of a carrier protein in vitro and in vivo, with mutations in either affecting the rate and extent of nuclear accumulation. The N-terminal NLS function is independent of the conventional NLS-binding importins (IMPs) and requires unidentified cytoplasmic transport factors, whereas the C-terminal NLS is recognized by IMPbeta. The SRY-R133W mutant shows reduced IMPbeta binding as a direct consequence of the sex-reversing C-terminal NLS mutation. Of the N terminal NLS mutants examined, SRY-R62G unexpectedly shows a marked reduction in IMPbeta binding, whereas SRY-R75N and SRY-R76P show normal IMPbeta binding, suggesting defects in the IMP-independent pathway. We conclude that SRY normally requires the two distinct NLS-dependent nuclear import pathways to reach sufficient levels in the nucleus for sex determination. This study documents cases of human disease being explained, at a molecular level, by the impaired ability of a protein to accumulate in the nucleus. PMID- 12764227 TI - Behavioral performance in adult honey bees is influenced by the temperature experienced during their pupal development. AB - To investigate the possible consequences of brood-temperature regulation in honey bee colonies on the quality of behavioral performance of adults, we placed honey bee pupae in incubators and allowed them to develop at temperatures held constant at 32 degrees C, 34.5 degrees C, and 36 degrees C. This temperature range occurs naturally within hives. On emergence, the young adult bees were marked and introduced into foster colonies housed in normal and observation hives and allowed to live out their lives. No obvious difference in within-hive behavior was noted between the temperature-treated bees and the foster-colony bees. However, when the temperature-treated bees became foragers and were trained to visit a feeder 200 m from the hive, they exhibited clear differences in dance performance that could be correlated with the temperatures at which they had been raised: bees raised at 32 degrees C completed only approximately 20% of the dance circuits when compared with bees of the higher-temperature group. Also, the variance in the duration of the waggle phase is larger in 32 degrees C-raised bees compared with 36 degrees C-raised bees. All other parameters compared across all groups were not significantly different. One-trial learning and memory consolidation in the bees raised at different temperatures was investigated 1 and 10 min after conditioning the proboscis-extension reflex. Bees raised at 36 degrees C performed as expected for bees typically classified as "good learners," whereas bees raised at 32 degrees C and 34.5 degrees C performed significantly less well. We propose that the temperature at which pupae are raised will influence their behavioral performance as adults and may determine the tasks they carry out best inside and outside the hive. PMID- 12764230 TI - Effects of thrombin inhibitor on thrombin-related signal transduction and cerebral vasospasm in the rabbit subarachnoid hemorrhage model. AB - BACKGROUND AND PURPOSE: Thrombin is activated in the cerebrospinal fluid (CSF) after a subarachnoid hemorrhage (SAH). However, the relationship between thrombin and cerebral vasospasm has not yet been fully established. The aim of this study was to investigate the possibility of thrombin as a causative factor for cerebral vasospasm and to delineate the signal transduction mechanism that results in thrombin-inducing sustained vasoconstriction in cerebral vasospasm. METHODS: In the SAH group, SAH was simulated by the 2-hemorrhage rabbit model. In the treatment group, antithrombin III (AT-III) was injected into the cisterna magna just before production of the SAH. CSF samples were obtained serially to measure d-dimer with latex photometric immunoassay. On day 4, the basilar artery was excised after perfusion-fixation. The degree of cerebral vasospasm was evaluated by measuring the cross-sectional area of each basilar arterial lumen, and the expression of mitogen-activated protein kinase (MAPK) in the vascular wall was examined with an immunohistochemical technique. RESULTS: In the treatment group, the value of d-dimer on day 4 was 0.83+/-0.07 microg/mL, which was statistically significantly lower than that in the nontreated SAH group (2.49+/-0.09 microg/mL, P<0.01). The cross-sectional area of the arterial lumen in the treatment group was 3.67x10(5)+/-1.58x10(4) square pixels, which was statistically significantly larger than that in the nontreated SAH group (2.60x10(5)+/-2.29x10(4) square pixels; P<0.01). MAPK was detected diffusely in the vascular smooth muscle cell layer in the nontreated SAH group, but it was absent in the treatment group. CONCLUSIONS: Inhibition of thrombin activity leads to amelioration of cerebral vasospasm and suppression of MAPK diphosphorylation. This suggests that thrombin and its related signal transduction, including the MAPK cascade, appear to play an important role in the pathogenesis of cerebral vasospasm after SAH. PMID- 12764228 TI - An autoregulatory loop controls peroxisome proliferator-activated receptor gamma coactivator 1alpha expression in muscle. AB - Skeletal muscle adapts to chronic physical activity by inducing mitochondrial biogenesis and switching proportions of muscle fibers from type II to type I. Several major factors involved in this process have been identified, such as the calcium/calmodulin-dependent protein kinase IV (CaMKIV), calcineurin A (CnA), and the transcriptional component peroxisome proliferator-activated receptor gamma coactivator 1alpha (PGC-1alpha). Transgenic expression of PGC-1alpha recently has been shown to dramatically increase the content of type I muscle fibers in skeletal muscle, but the relationship between PGC-1alpha expression and the key components in calcium signaling is not clear. In this report, we show that the PGC-1alpha promoter is regulated by both CaMKIV and CnA activity. CaMKIV activates PGC-1alpha largely through the binding of cAMP response element-binding protein to the PGC-1alpha promoter. Moreover, we show that a positive feedback loop exists between PGC-1alpha and members of the myocyte enhancer factor 2 (MEF2) family of transcription factors. MEF2s bind to the PGC-1alpha promoter and activate it, predominantly when coactivated by PGC-1alpha. MEF2 activity is stimulated further by CnA signaling. These findings imply a unified pathway, integrating key regulators of calcium signaling with the transcriptional switch PGC-1alpha. Furthermore, these data suggest an autofeedback loop whereby the calcium-signaling pathway may result in a stable induction of PGC-1alpha, contributing to the relatively stable nature of muscle fiber-type determination. PMID- 12764229 TI - Crystal structure of a bifunctional aldolase-dehydrogenase: sequestering a reactive and volatile intermediate. AB - The crystal structure of the bifunctional enzyme 4-hydroxy-2-ketovalerate aldolase (DmpG)/acylating acetaldehyde dehydrogenase (DmpF), which is involved in the bacterial degradation of toxic aromatic compounds, has been determined by multiwavelength anomalous dispersion (MAD) techniques and refined to 1.7-A resolution. Structures of the two polypeptides represent a previously unrecognized subclass of metal-dependent aldolases, and of a CoA-dependent dehydrogenase. The structure reveals a mixed state of NAD+ binding to the DmpF protomer. Domain movements associated with cofactor binding in the DmpF protomer may be correlated with channeling and activity at the DmpG protomer. In the presence of NAD+ a 29-A-long sequestered tunnel links the two active sites. Two barriers are visible along the tunnel and suggest control points for the movement of the reactive and volatile acetaldehyde intermediate between the two active sites. PMID- 12764231 TI - Anticoagulants versus antiplatelet agents for acute ischemic stroke. PMID- 12764232 TI - Upregulation of CD40-CD40 ligand (CD154) in patients with acute cerebral ischemia. AB - BACKGROUND AND PURPOSE: Inflammation and hypercoagulability contribute to the development of acute cerebral ischemia. Both can be mediated by the CD40 system. This study investigated whether the CD40 system and related mediators are upregulated in patients with transient ischemic attack (TIA) or stroke. METHODS: Seventeen patients with TIA, 60 patients with complete stroke, and 15 control subjects were investigated. CD154 and P-selectin were analyzed on platelets and CD40 on monocytes during and 3 months after acute cerebral ischemia by double label flow cytometry. Blood concentrations of soluble CD154 and monocyte chemoattractant protein-1 (MCP-1) were evaluated. RESULTS: Our main findings are as follows: (1) patients with acute cerebral ischemia showed a significant increase of CD154 on platelets and CD40 on monocytes compared with controls; (2) plasma levels of soluble CD154 were significantly higher in these patients; (3) these patients had significantly higher numbers of prothrombotic platelet monocyte aggregates; (4) the chemoattractant MCP-1 was significantly elevated in cerebral ischemia; and (5) at 3 months' follow-up, upregulation of CD154 still persisted in patients with previous acute cerebral ischemia. CONCLUSIONS: Patients with acute cerebral ischemia show upregulation of the CD40 system, which might contribute to the known proinflammatory, proatherogenic, and prothrombotic milieu found in these patients. PMID- 12764233 TI - Major risk factors for aneurysmal subarachnoid hemorrhage in the young are modifiable. AB - BACKGROUND AND PURPOSE: To identify risk factors for subarachnoid hemorrhage (SAH) and intracerebral hemorrhage, we designed a case-control study of men and women 18 to 49 years of age (the Hemorrhagic Stroke Project [HSP]). This report focuses on SAH. METHODS: Patients were recruited from 44 hospitals in the United States. Cases with SAH must have had a ruptured aneurysm documented by angiography or surgery. Two controls, identified by random digit dialing and matched to each patient for age, sex, race, and telephone exchange, were sought for each case subject. RESULTS: Between 1994 and 1999, 425 patients with SAH were enrolled in HSP, and 312 cases met the criteria for aneurysmal SAH. The present analyses also included 618 matched controls. Of the 312 cases, 66% were current cigarette smokers compared with 30% of controls (adjusted odds ratio [OR], 3.73; 95% CI, 2.67 to 5.21). Cocaine use within the previous 3-day period was reported by 3% of cases and no controls (bivariate exact OR, 24.97; 95% exact CI, 3.95 to infinity; adjusted estimate not calculable). Other independent risk factors in the multivariable model included hypertension (adjusted OR, 2.21; 95% CI, 1.48 to 3.29), low body mass index (OR, 1.59; 95% CI, 1.08 to 2.35), primary family history of hemorrhagic stroke (OR, 3.83; 95% CI, 1.73 to 8.46), caffeine in pharmaceutical products (OR, 2.48; 95% CI, 1.19 to 5.20), lower educational achievement (OR, 2.36; 95% CI, 1.44 to 3.87), and nicotine in pharmaceutical products (adjusted estimate not calculable). CONCLUSIONS: Aneurysmal SAH may be largely a preventable disease among the young and middle-aged because several prevalent risk factors can be modified by medication (eg, hypertension) or behavioral change (eg, cigarette smoking, cocaine use). The association of caffeine and nicotine in pharmaceutical products and aneurysmal SAH warrants further study. PMID- 12764234 TI - Editorial comment: Is MVP an MVP in ischemic cerebral events? PMID- 12764235 TI - Editorial comment: The fall and rise of lacunar infarction with carotid stenosis. PMID- 12764236 TI - Editorial comment: Stroke and the CD40-CD40 ligand system: at the hinge between inflammation and thrombosis. PMID- 12764237 TI - Investigation of an outbreak of E. coli O157 infections associated with a trip to France of schoolchildren from Somerset, England. AB - A retrospective cohort study was conducted after 2 confirmed and 8 probable cases of VTEC 0157 occurred in a schoolgroup from Somerset following a trip to France. The strain of the 2 confirmed cases was vero-cytotoxin type 2, phage type 34. Despite its low statistical power due to small numbers, this study suggested the most likely vehicle of infection was cucumber salad (crude RR=4.3, 95%CI [0.6 29], adjusted OR=107, 95%CI indeterminate, p=0.08). The cucumbers were purchased from Belgium but it was not possible to trace them back to source. The international nature of this outbreak emphasises the importance of close collaboration between organisations in the management of outbreaks, of ensuring international standards in food safety, and of agreeing a common standard in VTEC typing across Europe. PMID- 12764238 TI - Investigation of an unexplained skin disorder in a prison clinic in the Netherlands in 2002. AB - An unexplained outbreak of skin disorder with itching papules or pustules occurred among drugs body packers in a detention clinic in the Netherlands in October-November 2002. The descriptive cohort study suggests this outbreak could be associated with an expired batch of disposable linens, with an overall attack rate of 30% in the study population (n=118). No additionnal cases have been noted since this expired batch was replaced. PMID- 12764240 TI - Evaluation of Linearity for the Radiophotoluminescence Glass Dosimeter Based on Monochromatic X-rays. AB - Although low energy X-rays have been utilized for mammography, their safety in medical use is a matter of concern. Characteristics of the radiophotoluminescence glass dosimeter, GD-403, consisting of a glass element and filters, were investigated with respect to monochromatic X-rays obtained from a synchrotron radiation for personal monitoring of low energy photons. We focused on low energy X-rays ranging from 8 to 20 keV to study the linearity of the GD-403 response between photon fluence and dose equivalent. The GD-403 was placed on a tough water phantom and irradiated using an 11-15 mm x 0.1-7 mm beam for modulation of the photon fluence. The tough water phantom could be moved through a distance of 110-150 mm with a stepping motor. For the dose equivalent at 1cm depth (H1), 3mm (H3) and 70 &mgr;m (H70), the GD-403 showed sufficient linearities against the photon fluences in the energy regions of 8 to 20 keV, 13 to 20 keV and 13 to 20 keV, respectively. However, H3 and H70 did not provide sufficient linearities in the energy region of 8 to 12 keV. Moreover, we compared the result in this experiment with the value calculated from the absorbed dose of air using the mass absorption coefficient for the X-ray energy ranging from 10 to 20 keV. PMID- 12764241 TI - Determination of N(pp)(gas) and N(pp)(D) for a plane-parallel chamber. AB - The purpose of this work is to investigate the consistency of determining N(pp)(gas) and N(pp)(D) by using three independent calibration methods from the AAPM TG 39 and IAEA TRS 381 protocols: 1) calibration with a high-energy electron beam in a phantom; 2) in-phantom calibration in a (60)Co beam; and 3) in-air calibration in a (60)Co beam. The plane-parallel chamber considered was the PTW Markus and the comparisons were made against a calibrated PTW cylindrical Farmer type chamber 30001. The phantom material used for the electron beam and (60)Co in phantom methods was a solid water phantom (RW3). For the electron beam method, the nominal energies were 18 and 21 MeV. An acrylic buildup of 0.5 g/cm(2) thickness was used for the (60)Co in-air method. For each method, N(pp)(gas) and N(pp)(D) were obtained for the plane-parallel chamber as proposed by the AAPM TG 39 and IAEA TRS 381 protocols. The absorbed doses were measured along the central axis at a distance of 100 cm (SSD=100 cm) with 10 x 10 cm(2) field size at the depth of the maximum for each electron beam. The values of N(pp)(gas) by the three independent calibration methods agreed to within +/-0.6%. This meant that any of the methods would give a fairly good value. Similar results were obtained for N(pp)(D). In comparing the results for the electron beam method at energies of 18 and 21 MeV, the latter gave better agreement. The ratios of N(pp)(gas) and N(pp)(D) for the three methods were in agreement within 0.7%. The results for the absorbed dose intercomparison in the AAPM TG 39 and IAEA TRS 381 protocols showed that they agreed to within +/-0.7% which meant that any of the calibration methods and two different protocols would give an accurate result. PMID- 12764239 TI - Malaria in Austria 1990-2000. AB - In Austria, between 1990 and 2000, 924 travel related malaria cases were reported (mean = 84/year). No significant decreasing or increasing trends were observed. P. falciparum (n=517; 55.9%) accounted for the highest number of cases followed by P. vivax or ovale (n=321; 34.7%) and P. malariae (n=29; 2.2%). Most infections were contracted in highly endemic malaria regions (n=686; 74.2%) and most cases were reported from the largest counties: Vienna (n=336, 36.4%), Styria (n=156, 16.8%), and Lower Austria (n=151, 16.3%). Overall, 12 deaths occurred, most were caused by P. falciparum (n=9, 75%; case fatality rate: 1.9%). Data on chemoprophylaxis was available for 752 cases (81.4%) but only half of them (n=367, 48.8%) gave detailed information on the drug used. Data on compliance were obtained for only 45.4% of the cases, with only about 60% of patients completing the full course of prophylaxis. PMID- 12764242 TI - [Improvement of dose distributions at NIRS's 70 MeV proton eye treatment beam course] AB - In order to improve dose distributions at NIRS's 70 MeV proton eye treatment beam course, we introduced finer bar ridge filters, and examined the effects of range compensators. The pitch of new bar ridge filters was 5mm in contrast to 15mm pitch of old ones. A NC-machine recently available enabled this refinement. The spread out Bragg peak (SOBP) widths were 10, 15, 20 and 30 mm. The new ridge filters improved the field uniformity considerably. In the ridge filter design we assumed parallel beam condition in which the mono-energetic proton should proceed in parallel with the central axis, and bar ridges only changes the proton ranges. We searched empirically for the optimum wobbler radius in view of field flatness and depth dose distribution. Range shifter and compensator did not affect the field flatness and depth dose distribution at the optimum condition thus searched. We measured dose distribution in a phantom using a compensator of stairs-shape, which fairly modulated the beam. A 50% isodose line almost coincided with the compensator shape, and these results suggested that improvements of dose distributions should be possible using compensators. However width between 50% and 80% isodose lines depended on the thickness of phantom. This might be due to scattering in the compensator and suggests that it is necessary to calculate dose distribution taking account of such effects. PMID- 12764243 TI - [Usefulness of high energy radiation using copper filter on CR chest radiography] AB - The bone contrast on chest radiographs obtaind with computed radiography (CR, hereafter) is not decreased enough compaired with that for screen/film combination, even if we use a high tube voltage. This may be because the Exporsure Data Recognizer which can normalize the density and contrast of the chest images is performed. In order to increase the diagnostic accuracy for the lung disease,we propose to use a copper additional filter to decrease the bone contrast. Consequently, in order to have satisfied the bone contrast and the patient expousure dose, additional 0.3 mm copper filter and effective energy was 59.3 keV or more were required of the tube voltage of 140kV. Further, the effective energy of 140 kV tube voltage with a 0.7 mm copper, which is the maximum thickness of the additional filter within 50 m seconds in exporsure time, was 68 keV. Concluding these results, optimum range of additional copper filter is from 0.3 mm to 0.7 mm in chest high voltage radiography using CR. The bone contrast decreases sufficiently, if we use X-rays which have effective energy within this range. Our method is also useful to reduce patient dose. PMID- 12764244 TI - A Few Remarks on Photoluminescence Dosimetry with High Energy X-Rays. AB - The present study investigated the possibility of using PLD (photoluminescence dosimetry) for clinical radiotherapy of absorbed dose. The main parameters which provide desirable dosimetric properties of PLD in high energy x-rays for radiotherapy, such as linearity, fading, and random uncertainty, were studied and the following conclusions were reached. There is good linearity of PLD response to absorbed dose. The PLD response is independent of the x-ray energy and depth in the water phantom. PLD has unique properties, such as a small response dispersion among PLD glasses, reproducibility of measurements, and extremely low fading. Additionally, PLD has a high spatial resolution. It is suitable for exact dose measurement, especially for an extremely small field and/or in the high dose gradient region. Moreover, there is no need for a complicated washing process of PLD glass and PLD can be used to irradiate a bare glass in a water phantom. The PLD system is found to be a suitable dosimeter system for clinical radiotherapy. PMID- 12764245 TI - The Influence of DSA Examinations on Patient-absorbed Doses. AB - An exposure measurement experiment was performed to obtain frontal DSA, lateral DSA and fluoroscopic absorbed doses. In addition, the proportion of each dose to the total absorbed dose was obtained to estimate which influenced the patient absorbed dose the most among the three in diagnostic DSA examinations for the head and neck. Three correlation coefficients were obtained to evaluate the relationship between any two doses. As a result, the following four findings were obtained: 1. The mean proportion of each absorbed dose reached a high value in the order of frontal DSA absorbed dose > lateral DSA absorbed dose > fluoroscopic absorbed dose. 2. A strongly negative correlation between the proportions of frontal DSA absorbed dose and fluoroscopic absorbed dose was observed (r = 0.864, P < 0.01). Also, a slightly negative correlation between the proportions of lateral DSA absorbed dose and fluoroscopic absorbed dose was observed (r = 0.375, P < 0.01). However, no correlation between the proportions of frontal DSA absorbed dose and lateral DSA absorbed dose was observed (r = -0.143, P > 0.05). 3. The patient-absorbed doses, which were different from those of IVR examinations, were confirmed to be largely influenced by DSA radiographic doses. 4. The best method for the reduction of patient-absorbed doses was considered to be minimization of the frame number in DSA radiography required for diagnosis. PMID- 12764246 TI - [Fundamental Study on Heavy Ion CT Using Pencil Beam Scanning Method] AB - The feasibility of heavy ion computed tomography was investigated. A cylindrical polyethylene sample of 50 mm in diameter was used to establish the pencil beam scanning method and estimate the performance of the system. The sample was irradiated with carbon ions of 290 MeV/u and 400 MeV/u, and helium ions of 150 MeV/u. The residual beam energy was measured by a BGO scintillator in coincidence with two small plastic scintillators placed in front of and behind the sample to restrict the beam path and improve the spatial resolution in the CT reconstruction. The projection data were obtained by moving the sample only in the transverse direction because of its symmetrical structure. A Lucite sample with many holes of different diameters was used to demonstrate the spatial resolution of this CT system. From the reconstructed images the spatial resolution was estimated to be less than 2 mm under the conditions in this work. The electron density ratios of ethyl alcohol, water, and Lucite to polyethylene were obtained, which are in good agreement with the calculated values. PMID- 12764247 TI - Nonlinear Effect on Noise Power Spectrum for Quantum Mottle in Radiographs. AB - Rossmann's formula on the power spectrum of quantum noise in a radiograph is based on an assumption that the relation between the spatial fluctuations of optical density and the relative spatial fluctuations of light energy irradiated from the intensifying screen due to absorbed x-ray quanta is linear. The reasonability of the assumption is not theoretically obvious because the relative spatial fluctuation of light energy increases as the number of absorbed x-ray quanta decreases. Based on the shot-noise theory, the power spectrum of quantum noise is calculated taking account of the second order of the spatial fluctuation of light energy and the spectrum is expressed as a function of the gradient, the modulation transfer function of the screen-film system, and the average number of absorbed x-ray quanta. The term caused by the nonlinearity between the spatial fluctuations of optical density and light energy per unit area for the noise power spectrum is also estimated using the m-hits model for the emulsion and it is found that the effect due to the nonlinearity is little in practice. PMID- 12764249 TI - Another utilization of the density scaling theorem-attenuation models. PMID- 12764248 TI - Determination of overall perturbation factors for plane-parallel ionization chambers in electron beams. AB - Most dosimetry protocols recommend the use of plane-parallel chambers for dose determination in electron beams with energies below 10 MeV. The new IAEA TRS 381 (1997) protocol includes the overall perturbation factor p(Q) that consists of the in-scattering correction factor p(cav) (or P(repl)) and the wall correction factor p(wall) (or P(wall)). In this work, p(Q) for the commonly applied NACP, PTW/Roos and PTW/Markus plane-parallel chambers was determined experimentally. For the NACP plane-parallel chamber, p(Q) was obtained by comparison with a cylindrical Farmer chamber, while for the PTW/Roos and PTW/Markus chambers it was obtained by comparison with the NACP chamber. The values of p(Q) for these plane parallel chambers were measured as a function of mean electron energies E(z) from 1.7 MeV to 11.5 MeV. It was found that for the NACP and PTW/Roos chambers, p(Q) is independent of energy down to E(z) =1.7 MeV, while for the PTW/Markus chamber it shows a systematic and exponential drop of about 2% with decreasing energy down to E(z) = 2.7 MeV. However, the decrease of p(Q) for E(z) =1.7 MeV was not exponential. PMID- 12764250 TI - [Dosimetry study using an anthropomorphic head phantom for stereotactic radiosurgery] AB - A dosimetry study using an anthropomorphic head phantom (Radiosurgery Verification Phantom (RSVP)) and a micro-ionization chamber for stereotactic radiosurgery was performed. To study the characteristics of the micro-ionization chamber, tissue maximum ratios (TMRs), off-axis ratios (OARs) and output factors were measured and compared, using a Si semiconductor detector and the film method. The directional difference in sensitivity and the effect of the polarity of the micro-ionization chamber were measured. According to the results obtained, the measured values and the values obtained through calculation were in agreement within 6.3%. The difference was due to the larger size of the micro-ionization chamber as compared to the other method. Therefore we applied a correction for this size effect and obtained better agreement within 2%. On the basis of the results of the present study we were able to confirm the overall accuracy and establish a means of quality assurance (QA) for stereotactic radiosurgery. PMID- 12764251 TI - Inconsistencies of Microcalcification Specks in Phantoms Approved by the American College of Radiology for Mammography Accreditation. AB - PURPOSE: To quantitatively demonstrate inconsistencies in microcalcification specks in RMI-156 phantoms approved by the American College of Radiology (ACR) for use with their mammography accreditation program. MATERIALS AND METHODS: Signal intensities of the largest three specks (C1, 540 &mgr;m; C2, 400 &mgr;m; C3, 320 &mgr;m) were measured on images of 14 RMI-156 (version 4) phantoms. Three images were analyzed per phantom: a packaged image of the wax plate and two phantom images obtained at an optical density of 1.2 to 1.3. Phantom images were digitized using a VXR-12 scanner at a resolution of 85 &mgr;m and 8 bit/pixel. Speck signals were measured as the average pixel values using the NIH Image program (version 1.61). RESULTS: Signal intensities of specks measured on phantom images were consistent with those measured on wax-plate images, with an average correlation coefficient of 0.898 for the C1 groups and of 0.794 for the C3 groups. Overlap of speck sizes was found between different groups. Speck signals of some inserts showed considerable inconsistencies within a group. In some phantoms, signal intensities of specks in different groups overlapped to each other, and C1 and C2 groups were not clearly separated. In all 14 phantoms, speck signal intensities of the C3 group had a between-phantom variance which was larger than the within-phantom variance. CONCLUSION: Microcalcification specks in RMI-156 phantoms of the same version were found to be substantially inconsistent in terms of size and intrinsic signal intensity. As lack of uniformity in specks within a phantom and between phantoms is not appropriate as a standard of image quality control, further quality control is apparently needed in manufacturing RMI-156 phantoms and also in approving specific phantoms. PMID- 12764252 TI - Immunological detection of D-beta-aspartate-containing protein in lens-derived cell lines. AB - PURPOSE: Although the presence of biologically uncommon D-beta-aspartate (D-beta Asp) in lens protein is thought to be related to aging, we recently found this isomer in lens alphaA-crystallin from human newborns. The objective of this study was to examine whether D-beta-Asp occurs in protein from lens-derived cell lines. METHODS: We examined the expression of D-beta-Asp-containing protein in the lens derived cell lines alphaTN4-1 and N/N1003A, by western blot and immunoprecipitation analysis using a polyclonal antibody against Gly-Leu-D-beta Asp-Ala-Thr-Gly-Leu-D-beta-Asp-Ala-Thr-Gly-Leu-D-beta-Asp-Ala-Thr (peptide 3R), which corresponds to three repeats of positions 149-153 in human alphaA crystallin. The anti-peptide 3R antibody, prepared in a previous study, is a useful tool for investigating D-beta-Asp-containing peptides. RESULTS: Western immunoblot and immunoprecipitation analysis showed that a 50 kDa protein in N/N1003A cells was strongly immunoreactive with the anti-peptide 3R antibody. Antibodies against alphaA- and alphaB-crystallin also stained this protein. On the other hand, the alphaTN4-1 cell line only expressed proteins of about 20 kDa, which also reacted to antibodies against alphaA-crystallin and alphaB-crystallin. CONCLUSIONS: The results indicate that the N/N1003A cell line expressed a 50 kDa D-beta-Asp-containing protein, which may share a common amino acid sequence with alphaA- and alphaB-crystallin. PMID- 12764253 TI - A novel Xenopus SWS2, P434 visual pigment: structure, cellular location, and spectral analyses. AB - PURPOSE: The purpose of this study was to clone and characterize the green rod pigment in Xenopus laevis. METHODS: The cDNA for the Xenopus "green rod" pigment was cloned and sequenced from Xenopus retina mRNA by reverse transcription polymerase chain reaction and the 5' end cloned by rapid amplification of the cDNA ends. The cellular localization of the Xenopus opsin was determined by immunolabeling of flat-mounted retinas using a specific antibody against this opsin. Spectral properties of the expressed protein were determined by absorption spectroscopy using recombinant pigment. RESULTS: A novel Xenopus opsin cDNA containing a full-length coding region has been cloned and sequenced. The deduced amino acid sequence predicts a protein of 362 amino acids, forming 7 hydrophobic helices. Sequence analysis indicates that it belongs firmly to the SWS2 class of visual pigments and has 89%, 80%, and 75% amino acid sequence identity with bullfrog, tiger salamander, and newt SWS2 pigments, respectively. Staining of Xenopus retina with a Xenopus SWS2 opsin-specific polyclonal antibody demonstrated that the SWS2 pigment is expressed in green rods. After expression in COS cells, reconstitution with 11-cis retinal, and purification, the SWS2 pigment exhibits an absolute absorption maximum of 434 nm Thus, the name "SWS2, P434" was assigned for this opsin. The pigment decays rapidly in hydroxylamine in the dark, unlike the red rod pigment, rhodopsin. CONCLUSIONS: A novel green rod opsin cDNA has been cloned and sequenced from the retina of adult Xenopus laevis, which encodes a protein belonging to the SWS2 group of opsins. The expressed opsin possesses cone-opsin-like properties although it was identified only in the Xenopus green rod cells. PMID- 12764254 TI - Amyloid-beta is found in drusen from some age-related macular degeneration retinas, but not in drusen from normal retinas. AB - PURPOSE: Age-related macular degeneration (AMD) is the most common cause of irreversible vision loss in the elderly. Increased understanding of the pathogenesis is necessary. Amyloid-beta (Abeta), a major extracellular deposit in Alzheimer's disease plaques, has recently been found in drusen, the hallmark extracellular deposit in AMD. The goal of this study was to characterize the distribution and frequency of Abeta deposits in drusen from AMD and normal post mortem human retinas to gain additional insight about the potential role of Abeta in AMD patho genesis. METHODS: Immunocytochemistry was performed with three Abeta antibodies on sections from 9 normal and 9 AMD (3 early, 3 geographic atrophy, 3 exudative AMD) retinas. Five sections from each eye were evaluated. Abeta positive deposits in drusen were identified using epifluorescence and confocal microscopy. Antibodies were pre-adsorbed with Abeta peptide to verify specificity. Some sections were stained with PAS-hematoxylin to aid in evaluation of morphology. RESULTS: To test and optimize immunocytochemistry, Abeta was detected in amyloid plaques from Alzheimer's brains. Abeta label was blocked by pre-adsorption of antibody with Abeta peptide, verifying specificity. Four of the 9 AMD retinas and none of the 9 normal retinas had Abeta positive drusen. Two of the early AMD eyes had a few A[beta] positive drusen, each with a few Abeta containing vesicles, and 2 of the geographic atrophy (GA) eyes had many Abeta positive drusen with many Abeta containing vesicles. CONCLUSIONS: Abeta was present in 4 of 9 AMD eyes. Within these eyes, Abeta localized to a subset of drusen. None of the 9 normal eyes surveyed, some of which had small drusen, were A beta positive. Abetapositive vesicles were most numerous in GA eyes at the edges of atrophy, the region at risk for further degeneration. These results suggest that Abeta in drusen correlates with the location of degenerating photoreceptors and retinal pigment epithelium (RPE) cells. Further work will be necessary to determine whether Abeta deposition in drusen may contribute to or result from retinal degeneration. PMID- 12764255 TI - Presidential address: carotid endarterectomy, under attack again! PMID- 12764256 TI - Durability of aortouniiliac endografting with femorofemoral crossover: 4-year experience in the Evt/Guidant trials. AB - OBJECTIVES: We evaluated mid-term results of the multicenter EVT/Guidant aortouniiliac endograft (AI) trial and ascertained the durability of this endovascular technique in patients unable to undergo standard bifurcated endografting. METHODS: From November 1996 to December 1998, 121 patients were enrolled to receive the AI device on the basis of complex iliac artery anatomy contraindicating bifurcated endografting. Clinical data were centrally collected, and radiographic data were evaluated by core facility. RESULTS: AI placement was technically successful in 113 of 121 patients. At operation, patients who underwent AI had significantly more arrhythmias, congestive heart failure, and peripheral occlusive disease (P <.05) compared with patients who underwent open aneurysmorrhaphy in the EVT/Guidant trials, indicating comorbid features in this anatomic cohort. Distal AI attachment was performed to the external iliac artery in 40 (36%) patients. Median follow-up was 38 months. In the AI group, overall aneurysm diameter decreased over the duration of study from 54.4 +/- 9.6 mm to 44.4 +/- 16.4 mm (P =.004). At 24 and 36 months after repair, reduction in aneurysm size was associated with absence of endoleak (P =.003 and P =.008, respectively). Aneurysms shrunk or remained stable in 109 (96.5%) patients. Endoleak was identified in 52.3% of patients at discharge, and at follow-up in 30.9% at 1 year, 34.8% at 2 years, 28.6% at 3 years, and 30.4% at 4 years. Type II endoleak predominated. Leak from failure to completely occlude contralateral iliac flow accounted for 8 of 58 endoleaks (13.8%) at discharge. Sixteen patients (14.2%) underwent postoperative endoleak treatment; in one of these patients open conversion was necessary at 20 months. Post-procedure thigh or buttock claudication developed in 16 patients (14.2%). Thirteen patients (81.3%) had either distal attachment in the external iliac artery or contralateral type IIA occlusion. Fifteen patients (13.3%) required intervention because of reduced limb flow; one of these patients underwent open conversion at 27 months, and another underwent axillofemoral grafting at 28 months. Device migration was confirmed in 2 (1.8%) patients, without current clinical sequelae. Whereas no femorofemoral graft thromboses occurred, graft infection developed in 3 patients (2.6%). During follow-up, aneurysm in 2 patients ruptured. Late death occurred in 41 patients (36.3%). Twenty-four patients (58.5%) died of cardiopulmonary disease; one death was endograft-related after aneurysm rupture; and one death was related to femorofemoral bypass infection. Actuarial survival was 78.4% (95% confidence interval [CI], 71%-86%) at 2 years and 63.4% (95% CI, 54%-73%) at 4 years. CONCLUSIONS: In patients with significant comorbid conditions and complex iliac anatomy unfavorable for bifurcated endografting, AI with femorofemoral bypass grafting is safe and effective. In most patients this endovascular option provides satisfactory mid-term results. PMID- 12764257 TI - Incision and abdominal wall hernias in patients with aneurysm or occlusive aortic disease. AB - INTRODUCTION: Patients undergoing midline incision for abdominal aortic reconstruction appear to be at greater risk for postoperative incision hernia compared with patients undergoing celiotomy for general surgical procedures. Controversy exists as to whether incidence of abdominal wall hernia and increased risk for incision hernia is higher in patients with abdominal aortic aneurysm (AAA) than in patients operated on because of aortoiliac occlusive disease (AOD). We conducted a prospective multi-institutional study to assess frequency of incision hernia after aortic surgery through a midline laparotomy and of previous abdominal wall hernia. METHODS: Patients with AAA (n = 177) or AOD (n = 82) from three major institutions were prospectively enrolled in the study and examined. Data collected included demographic data, cardiopulmonary risk factors, smoking status, history of previous or current abdominal wall hernia (incision, inguinal, umbilical, femoral), previous midline incision, suture type, and postoperative complications. At a minimum of 6 months after laparotomy, patients were evaluated clinically for a new incision hernia. Differences were tested with the unpaired t test, X(2) test, or Fisher exact test, and multiple logistic regression was used to control for confounding variables. RESULTS: Mean follow-up of the cohort was 32.8 +/- 2.3 months. Rate of abdominal wall hernia and inguinal hernia in patients with AAA versus AOD was 38.4% versus 11% (P =.001) and 23.7% versus 6.1% (P =.003), respectively. Rate of postoperative incision hernia in patients with AAA was 28.2%, and in patients with AOD was 11.0% (P =.002). Adjusting for age, smoking, chronic obstructive pulmonary disease, body mass index, diabetes, bowel obstruction, and suture type, patients with AAA had almost a ninefold risk for postoperative incision hernia formation (odds ratio [OR], 8.8; P =.0049). CONCLUSION: Compared with patients with AOD, patients with AAA have a higher frequency of abdominal wall hernia and inguinal hernia, and are at significant increased risk for development of incision hernia postoperatively. The higher frequency of hernia formation in patients with AAA suggests the presence of a structural defect within the fascia. Further studies are needed to delineate the molecular changes of the aorta and its relation to the abdominal wall fascia. PMID- 12764258 TI - Management of endoleak after endovascular aneurysm repair: cuffs, coils, and conversion. AB - OBJECTIVE: The effectiveness of endovascular treatment of abdominal aortic aneurysm (AAA) may be limited by persistent perfusion of the aneurysm sac (endoleak). Endoleak that results in persistent systemic pressurization of the aneurysm or in continued AAA expansion is believed to require treatment to prevent rupture. This report describes the results of three techniques used to treat endoleak. METHODS: Endovascular repair of AAA was performed in 597 patients between January 1996 and September 2002. Seventy-three endoleaks that required treatment developed in 70 patients (11.7%). These involved the graft attachment site (type I) or the graft junction site (type III) or originated from collateral side-branch vessels (type II) and were associated with an increase in aneurysm size. Endoleak type was confirmed at angiography in all cases. Average time between the initial endovascular procedure and endoleak treatment was 14.5 +/- 5.7 months. The techniques used for endoleak treatment were deployment of an endovascular extension graft or cuff (n = 44), coil embolization (n = 24,) and conversion to conventional open repair (n = 5). Configurations of endovascular grafts in which endoleak developed were bifurcated (n = 44), aortouniiliac (n = 15), and aortoaortic-tube (n = 11). Mean follow-up after endoleak treatment was 24.5 +/- 12.2 months (range, 1-60 months). RESULTS: Endovascular extension grafts or cuffs were used to treat 41 attachment site endoleaks and 3 graft junction endoleaks, with overall technical success rate of 97%. Embolic coils were used to treat 16 retrograde side-branch endoleaks and 8 attachment site endoleaks, with overall technical success rate of 87%. Conversion to open surgery was performed in 4 patients with attachment site endoleaks and 1 patient with a graft junction site endoleak, and was successful in all cases. After endoleak treatment, aneurysm size decreased (>5 mm) in 38% of patients, stabilized in 58% of patients, and increased (>5 mm) in 4% of patients. Major morbidity occurred in 7.0%, with no perioperative deaths. CONCLUSIONS: Endovascular extension grafts, coil embolization, and conversion to open surgery each may be used to effectively repair endoleak. Selection of the treatment method used is determined by the anatomic characteristics of the endoleak and the patient's ability to tolerate conventional repair. Conversion to open repair was uniformly successful. Deployment of an extension cuff was successful when complete closure of the endoleak was achieved. Embolic coils were effective for retrograde endoleaks and provided stabilization of AAA size in selected patients with attachment site endoleaks in limited follow-up. PMID- 12764259 TI - Effect of suprarenal versus infrarenal aortic endograft fixation on renal function and renal artery patency: a comparative study with intermediate follow up. AB - PURPOSE: Suprarenal fixation of aortic endografts appears to be a safe option in patients with a short or conical proximal aortic neck. However, concern persists regarding the long-term effect on renal function when renal artery ostia are crossed by the uncovered stent. We investigated the effect of suprarenal versus infrarenal endograft fixation on renal function and renal artery patency after endovascular aortic aneurysm repair. METHODS: Records of 91 patients who underwent endovascular aortic aneurysm repair with a modular bifurcated stent graft between November 1999 and January 2002 were reviewed retrospectively. Two patients receiving dialysis because of chronic renal failure were excluded. Infrarenal fixation was used in 57 patients (group 1), and suprarenal fixation was used in 32 patients (group 2). In two patients in group 1 a Gianturco Z stent was inserted transrenally because of intraoperative proximal type I endoleak, and data for these patients were excluded from analysis. Follow-up evaluation was performed at 1, 6, and 12 months, and yearly thereafter, and included clinical assessment, measurement of serum creatinine concentration (SCr), and computed tomography angiography, per standard protocol. Median follow-up was 12 months (range, 1-36 months). RESULTS: There was no statistically significant difference in patient demographic data, aneurysm size, or preoperative risk factors. Median SCr was significantly higher in group 2 (suprarenal fixation) than in group 1 (infrarenal fixation) preoperatively (1.2 mg/dL [range, 0.6-2.3 mg/dL] vs 0.9 mg/dL [range, 0.6-1.9 mg/dL], P =.008) and at 1 month postoperatively (1.1 mg/dL [range, 0.8-5.6 mg/dL] vs 1.0 mg/dL [range, 0.6-2.1 mg/dL], P =.045). There was a significant increase in median SCr in both groups at 1 month postoperatively (group 1, 1.0 mg/dL [range, 0.6-2.1 mg/dL], P =.05; group 2, 1.1 mg/dL [range, 0.8-5.6 mg/dL] [mean SCr, 1.35 mg/dL vs 1.15 mg/dL, respectively], P <.05). In group 1 SCr was increased significantly at 6 and 12 months (P <.001), whereas in group 2 SCr also increased at 6 and 12 months, but not significantly. The change in SCr over time was not significantly different between the two groups. In two of 32 patients in group 2, renal artery occlusion developed, associated with perfusion defects in renal parenchyma and persistently elevated SCr. Analysis of renal artery patency did not demonstrate any association between patency and treatment. No patient developed hypertension during follow-up. CONCLUSIONS: Suprarenal endograft fixation does not lead to significant renal dysfunction, and renal artery occlusion is uncommon within 12 months. A larger study with longer follow-up is essential to determine overall effects on renal function and renal artery patency. PMID- 12764260 TI - Surgical treatment of intact thoracoabdominal aortic aneurysms in the United States: hospital and surgeon volume-related outcomes. AB - OBJECTIVE: Surgical treatment of intact thoracoabdominal aortic aneurysm (TAAA) is crucial to prevent rupture but is associated with high perioperative mortality. We tested the hypothesis that provider volume of surgical treatment of TAAA is an important determinant of operative outcome. Patients and methods Clinical information regarding repair of intact TAAA in 1542 patients from 1988 to 1998 was obtained from the Nationwide Inpatient Sample (NIS), a stratified discharge database of a representative 20% of US hospitals. Demographic data included age, sex, race, nature of admission, and comorbid conditions. Annual hospital volume of TAAA treated was grouped into terciles and defined as low (LVH; 1-3 cases [median, 1]), medium (MVH; 2-9 cases [median, 4]), or high (HVH; 5-31 cases [median, 12]). Annual surgeon volume was defined as low (LVS; 1-2 cases [median, 1]) or high (HVS; 3-18 cases [median, 7]). The primary outcome measure was in-hospital postoperative mortality. Secondary outcome measures included length of stay, and cardiac, pulmonary, and renal complications. Adjusted and unadjusted analyses were conducted. RESULTS: Overall mortality was 22.3%. Mortality improved over time. LVH and HVH differed in mortality rates (27.4% vs 15.0%; P <.001). Mortality between LVS and HVS also differed significantly (25.6% vs 11.0%; P <.001). When controlling for patient demographic data, comorbid conditions, and postoperative complications, both hospital and surgeon volume were significant predictors of mortality for intact TAAA repair (LVS: odds ratio [OR] 2.6, P <.001; LVH: OR 2.2, P <.001; and MVH: OR 1.7, P =.004). CONCLUSIONS: Greater hospital and surgeon TAAA treatment volumes contribute to better outcome. Given the relative high perioperative mortality associated with TAAA repair, regionalization of care to high-volume providers with consistently lower postoperative mortality deserves consideration by patients, physicians, and health care planners. PMID- 12764261 TI - Early results of stent-grafting to treat diffuse aortoiliac occlusive disease. AB - INTRODUCTION: Diffuse aortoiliac occlusive disease or isolated external iliac artery occlusion >or=10 cm long typically are treated with surgical bypass grafting because of limited durability of stent placement. We evaluated the results of stent-graft placement as an option for treatment of these diseases. METHODS: Demographic data for patients undergoing stent-grafting in one or more iliac artery segments were recorded. Technical, clinical, and hemodynamic success, and aortoiliac primary and secondary patency were analyzed with Society for Vascular Surgery/American Association for Vascular Surgery criteria. RESULTS: Thirty-four consecutive patients underwent stent-graft treatment because of rest pain (65%) or tissue loss (35%). Mean patient age was 63 years, and 38% of patients were women. Ninety-one percent of patients had hypertension, 71% had coronary artery disease, 21% had renal insufficiency (serum creatinine > 2.0 mg/dL), and 26% had diabetes; 71% were active smokers. TransAtlantic Inter Society Consensus C or D disease was present in 85% of patients, complete common or external iliac artery occlusion was present in 41%, and external iliac artery disease requiring treatment was present in 94% of patients. Mean lesion length was 13.7 +/- 8 cm. Technical, hemodynamic, and clinical success was achieved in all patients in whom the lesion could be crossed with a wire. Ankle-brachial index increased from.30 +/-.03 to.59 +/-.04. Self-expanding stent grafts were used (Wallgraft, Boston Scientific, Boston, Mass, in 88% of patients; Viabahn, W. L. Gore, Flagstaff, Ariz, in 12% of patients). Concomitant common femoral endarterectomy was performed in 53% of patients. At 12 months, primary patency was 70% and primary assisted patency was 88%. Four stent grafts became occluded because of distal external iliac artery or proximal common femoral artery disease, which required subsequent common femoral endarterectomy and either external iliac artery stent grafting or extraanatomic bypass grafting. Eighty percent (four of five) of primary patency failures were in patients who did not undergo concomitant common femoral endarterectomy at initial stent graft placement. CONCLUSION: Early results of stent-graft placement to treat diffuse aortoiliac occlusive disease appear better than our recent experience with stenting alone. Concomitant common femoral endarterectomy or better assessment of femoral disease may improve durability. PMID- 12764262 TI - Infrainguinal revascularization after renal transplantation. AB - BACKGROUND: Although evidence suggests that end-stage renal disease is associated with poor limb salvage and patient survival after arterial revascularization, little is known about the effect of renal transplantation. We analyzed the outcome in patients with renal transplants who underwent infrainguinal bypass procedures. METHODS: Data prospectively entered into our vascular registry were reviewed for all patients who underwent lower extremity bypass procedures from January 1, 1990, through January 31, 2002. Sixty patients were identified who had a functioning renal allograft at infrainguinal revascularization. Kaplan-Meier survival curves were generated for limb salvage, patency, and patient survival and were compared with the Mantel-Cox log- rank test. RESULTS: Sixty patients (40 men, 20 women; mean age, 47.1 years) underwent 76 bypass procedures in 71 limbs. Preoperative demographic data included diabetes (59 of 60 patients, 98.3%), coronary artery disease (26 of 60 patients, 43.3%), and preoperative serum creatinine concentration (SCr) greater than 2.0 mg/dL (9 of 60 patients, 11.7%). Mean follow-up was 25.1 months. Overall major complication rate was 11.8%, and 30 day mortality rate was 1.3%. Survival was 93.3% at 1 year and 66.6% at 5 years. Limb salvage was 87% at 1 year and 78% at 5 years. Primary graft patency was 78% at 1 year and 44% at 5 years. Preoperative SCr less than or equal to 2.0 mg/dL was associated with improved overall patient survival (5-year survival, 73.4% vs 37.5%; P =.01, log-rank test). Limb salvage and patency rates were not significantly affected by preoperative SCr greater than 2.0 mg/dL. CONCLUSIONS: Lower extremity bypass can be performed safely and effectively in patients who have undergone renal transplantation. However, the importance of a well functioning renal allograft at surgery is demonstrated by marked improvement in patient survival. PMID- 12764263 TI - Blinded comparison of preoperative duplex ultrasound scanning and contrast arteriography for planning revascularization at the level of the tibia. AB - PURPOSE: We examined whether preoperative duplex ultrasound scanning (DU) could replace contrast material-enhanced arteriography (CA) in selecting the recipient artery of tibial or peroneal artery bypass grafts. METHODS: In patients who underwent tibial or peroneal artery bypass grafting because of critical ischemia, images were obtained of the lower extremity arterial circulation with both DU and CA. Vascular surgeons, blinded to the operation performed, reviewed either DU or CA images for arterial visualization and patency. The tibial or peroneal artery best suited to receive the bypass graft was selected by surgeons using only data from either DU or CA images. This selection was compared with the artery actually used at bypass surgery. RESULTS: Preoperative DU and CA data for 40 lower extremities in 38 patients undergoing bypass grafting at the level of the tibia provided 110 arteries: 38 anterior tibial arteries, 32 peroneal arteries, and 40 posterior tibial arteries. Ten arteries (8 peroneal, 2 anterior tibial) were not identified with DU, and 1 artery (anterior tibial) was not identified with CA. DU enabled prediction of the artery actually used in 88% of patients (35 of 40), whereas CA enabled prediction of the artery actually used in 93% of patients (37 of 40; P =.59). Duplicate findings at DU and CA enabled selection of 85% of arteries actually used (95% confidence interval, 71%-93%). Arteries used for bypass grafting had significantly higher peak systolic velocity (35 cm/s vs 25 cm/s; P =.04), higher end-diastolic velocity (15 cm/s vs 9 cm/s; P =.005), and greater diameter (2.4 mm vs 1.7 mm; P =.003) compared with arteries not selected for bypass grafting. CONCLUSION: Findings at DU and CA typically agree when used to select tibial or peroneal arteries for bypass grafting. With DU there is occasional difficulty in identification of the peroneal artery, but selection of the actual artery used is accurate. Peak systolic velocity, end-diastolic velocity, and diameter characteristics correlate with arteriographic criteria for tibial bypass target artery selection. If DU enables adequate identification of a target artery for bypass grafting, and especially if the peroneal artery is seen, findings at CA are not likely to alter bypass execution. PMID- 12764264 TI - Preoperative risk factors for carotid endarterectomy: defining the patient at high risk. AB - PURPOSE: The efficacy of carotid endarterectomy (CEA) for prevention of stroke has been demonstrated in randomized trials; however, the optimal approach in patients excluded from these trials or who have other significant comorbid conditions remains controversial, particularly with the advent of percutaneous interventions. We examined the influence of putative risk factors on outcome of CEA in a single-center experience. METHODS: A retrospective analysis of 1370 consecutive CEA performed from 1990 to 1999 was undertaken. Preoperative risk factors examined included age older than 80 years, congestive heart failure, chronic obstructive pulmonary disease, renal failure (serum creatinine concentration > 2.0 mg/dL), contralateral carotid artery occlusion, recurrent ipsilateral carotid artery stenosis, ipsilateral hemispheric symptoms within 6 weeks, and recent coronary bypass grafting (CABG). The Fisher exact test was used to identify baseline variables associated with perioperative (30 days) risk for stroke or death. Multivariate analysis with Poisson regression was used to study the effect of all univariate criteria in combination. RESULTS: In the overall cohort, there were 32 adverse events (2.3%), including 11 deaths (0.8%), 6 disabling strokes (0.4%), and 10 nondisabling strokes (0.7%). There was no significant difference in incidence of perioperative stroke or death between patients with one or more risk factors (n = 689) and those with no risk factors (low risk, n = 681). Thirty-day mortality was significantly greater in patients with two or more risk factors compared with patients with no risk factors (2.8% vs 0.3%; P =.04), but no significant difference was noted in perioperative stroke rate (2.3% vs 1.0%). Univariate analysis demonstrated that contralateral carotid occlusion (n = 75) was the only significant predictor of adverse outcome (5 events, 6.7%) among the variables tested; this was confirmed with multivariate analysis (relative risk, 4.3; 95% confidence interval, 1.2-12.3; P =.01). Five year survival for patients with two or more risk factors was notably diminished compared with that for patients with no risk factors (38.7% +/- 5.9% vs 75.0% +/- 2.6%; P <.001). Contralateral occlusion was also associated with reduced 5-year survival (38 +/- 11% vs 67 +/- 2%; P <.004). CONCLUSION: CEA can be safely performed in patients deemed at high risk, including those aged 80 years or older and others with significant comorbid conditions, with combined stroke and mortality rates comparable to those found in randomized trials, ie, the Asymptomatic Carotid Atherosclerosis Study and the North American Symptomatic Carotid Endarterectomy Trial. Contralateral occlusion may be a predictor for moderately increased perioperative risk and for reduced long-term survival. Caution may be warranted in asymptomatic patients with multiple risk factors, in whom presumed long-term benefit of CEA may be compromised by markedly reduced 5 year survival. PMID- 12764265 TI - Predictive factors and clinical consequences of proximal aortic neck dilatation in 230 patients undergoing abdominal aorta aneurysm repair with self-expandable stent-grafts. AB - OBJECTIVE: Several studies have suggested that proximal aortic neck dilatation (AND) is a frequent event after balloon-expandable endografting. Yet few data are available on AND after repair with self-expandable stent grafts. To investigate incidence, predictive factors, and clinical consequences of AND, computed tomography (CT) scans obtained at intervals during follow-up of 230 patients who had undergone endoluminal abdominal aortic aneurysm (AAA) repair with self expandable stents were reviewed. SUBJECTS: Between April 1997 and March 2001, 318 patients underwent endoluminal AAA repair with a self-expandable endograft at our unit. CT scans obtained at 1 and 12 months after surgery and yearly thereafter were prospectively stored in a computer imaging data base. Two hundred thirty patients were available for minimum 1-year assessment. Two vascular surgeons with tested interobserver agreement reviewed 686 CT scans. Diameter of the proximal aortic neck was measured as the minor axis of the first CT section that contained at least half of the proximal portion of the endograft. For endografts with suprarenal attachment the first scan below the lowest renal artery was considered. Diameter change of 3 mm or more between the CT scan at 1 month and subsequent evaluations was defined as AND. Nine possible independent predictors of AND were analyzed with Cox regression analysis. RESULTS: Median follow-up was 24 months (range, 12-54 months). In 2 patients, AAA ruptured during follow-up. CT scans for 65 patients (28%) showed AND. Thirteen patients with AND (5.6%) underwent repeat intervention, including positioning of the proximal cuff in 8 patients and late conversion to open repair in five patients. Of the nine variables examined with multivariate analysis, only 3, ie, presence of neck circumferential thrombus (hazard ratio [HR], 2.51; 95% confidence interval [CI], 1.26-5.01; P =.008), preoperative proximal neck diameter (HR, 1.21; 95% CI, 1.07 135; P =.001), and preoperative AAA diameter (HR, 1.03; 95% CI, 1.00-1.06; P =.046) were positive independent predictors of AND, whereas the other 6, ie, neck angulation more than 60 degrees, neck length, suprarenal fixation, oversizing more than 15%, endoleak at 30 days, and increased AAA diameter during follow-up, showed no significant correlation. Probability of AND at 48 months was 59 +/- 6.1 at analysis with the Kaplan-Meier method. CONCLUSIONS: AND is a frequent sequela of endoluminal repair in the mid-term. Severe AND developed in a small percentage of our patients, compromising integrity of AAA repair. Patients with large aneurysms and aortic necks and patients with aortic neck circumferential thrombus are at high risk for aortic neck enlargement after endoluminal repair of AAA. PMID- 12764266 TI - Disparate outcome after endovascular treatment of small versus large abdominal aortic aneurysm. AB - OBJECTIVES: The size of an abdominal aortic aneurysm is the most important parameter for determining whether repair is appropriate. This decision, however, must be considered in the context of long-term outcome of treatment, balancing risk for rupture with mortality from the initial procedure and all subsequent secondary procedures necessary when durability is not ideal. Information on the results of endovascular repair of small versus large aneurysms has not been available. METHODS: Preoperative imaging studies and postoperative outcome were assessed in 700 patients who underwent endovascular repair of abdominal aortic aneurysm over 6 years at a single institution. Patients were divided into two groups: 416 patients (59.4%) with aneurysms smaller than 5.5 cm in diameter and 284 patients (40.6%) with aneurysms 5.5 cm or larger in diameter. Outcome variables were assessed with the Kaplan-Meier method and the log-rank test. RESULTS: Patients with small and large aneurysms were comparable with regard to all baseline parameters assessed, with the single exception of a small increase in age (2.3 years) in patients with large aneurysms (P =.031). While there were no differences in rate of type II endoleaks, mid-term changes in sac diameter, or aneurysm rupture between the two groups, at 24 months patients with large aneurysms had more type I leaks (6.4% +/- 2.3% vs 1.4% +/- 0.6%; P =.011), device migration (13% +/- 4.0% vs 4.4% +/- 1.8%; P =.006), and conversion to open surgical repair (8.2% +/- 3.2% vs 1.4% +/- 1.1%; P =.031). Of greatest importance, at 24 months patient survival was diminished (71% +/- 4.6% vs 86% +/- 2.8%; P <.001) and risk for aneurysm-related death was increased (6.1% +/- 2.6% vs 1.5% +/- 1.0%; P =.011) in the group with large aneurysms. CONCLUSIONS: Outcome after endovascular repair of abdominal aortic aneurysm depends on size; results appear inferior in patients with larger aneurysms. These differences attain importance when choosing between observation and repair, balancing risk for rupture against size-dependent outcome. PMID- 12764267 TI - Blood coagulation and fibrinolytic response after endovascular stent grafting of thoracic aorta. AB - OBJECTIVE: Thrombosis is common in aneurysms immediately after stent-grafting, because of exclusion from systemic blood flow. We studied changes in blood coagulation and the fibrinolytic system in patients with thoracic aortic aneurysm or dissection after stent-grafting to examine risk for consumption coagulopathy. METHODS: Thirty-one thoracic aortic aneurysms were treated with stent-grafting (aneurysm group), and 29 aortic dissections were treated with entry closure with stent-grafting (dissection group). The stent-graft was constructed from a self expanding Z stent and thin-walled woven polyester fabric. Platelet count, fibrinogen, antithrombin III (AT III), and thrombin-AT III complex were assayed as markers of coagulation. Plasminogen, alpha(2)-plasmin inhibitor, alpha(2) plasmin inhibitor-plasmin complex, fibrin degradation products fragment E (FDP E), and fibrin degradation products d-dimer were monitored as markers of fibrinolysis. Blood samples were collected before surgery and on postoperative days 1, 3, 7, and 14. RESULTS: In both groups platelet count significantly decreased on postoperative days 1 and 3, and increased on postoperative day 14. AT III significantly decreased on postoperative day 1, but recovered after postoperative day 7. FDP-E significantly increased on postoperative day 1 in both groups. There was significant correlation of aneurysm diameter with alpha(2) plasmin inhibitor-plasmin complex, fibrin degradation products, and d-dimer in the dissection group on postoperative day 1. CONCLUSIONS: Activation of coagulation and fibrinolysis was observed after stent-grafting to treat thoracic aortic aneurysm and aortic dissection. However, no patients exhibited consumption coagulopathy postoperatively. Therefore we believe there is little risk for consumption coagulopathy after stent-grafting. PMID- 12764268 TI - The impact of patient age and aortic size on the results of aortobifemoral bypass grafting. AB - OBJECTIVES: On the basis of the widespread belief that aortobifemoral bypass (ABF) represents the optimal mode of revascularization for patients with diffuse aortoiliac disease, vascular surgeons are often aggressive about its application in young adults. We undertook this retrospective evaluation of ABFs performed from 1980 to 1999 to determine whether the results justify this approach. Patients of less than 50 years of age (n = 45) were compared with those aged 50 to 59 years (n = 93) and those aged more than 60 years (n = 146). RESULTS: Younger patients were more likely to undergo operation for claudication than were older patients (72% versus 59% and 55%; P <.04). Younger patients were significantly more likely to be smokers (87%) but less likely to have diabetes, hypertension, or cerebrovascular disease. Bypasses were constructed in an end-to end fashion in 71.1% of patients of less than 50 years versus 68.8% and 71.2% of older patients (P = not significant). The mean diameter of aortic grafts was significantly smaller in younger patients (14.6 mm) than in older patients (15.6 mm and 15.5 mm; P <.01). The need for a subsequent infrainguinal reconstruction was highest in the youngest patients (24% versus 17% and 7%; P <.01). Surgical mortality rates were low in all groups (0%, 1%, and 2.0% for increasing age groups; P = not significant). Five-year primary and secondary patency rates increased significantly with each increase in age interval: 5-year primary patency rate: less than 50 years, 66% +/- 8%; 50 to 59 years, 87% +/- 5%; more than 60 years, 96% +/-2% (P <.05 for all comparisons). Five-year secondary patency rates were: less than 50 years, 79% +/- 7%; 50 to 59 years, 91% +/- 4%; more than 60 years, 98% +/- 2% (P <.05 for all comparisons). Five-year survival rate was comparable in all three groups: less than 50 years, 93% +/- 5%; 50 to 59 years, 92% +/- 4%; more than 60 years, 87% +/- 4% (P = not significant). CONCLUSION: Increased virulence of aortic disease, smaller aortic size, and more progressive infrainguinal disease may all negatively impact the results of ABF in younger patients. Although 5-year results are acceptable, increased caution is warranted in the routine application of ABF in young patients without limb threatening ischemia. PMID- 12764269 TI - Screening for asymptomatic internal carotid artery stenosis and aneurysm of the abdominal aorta: comparing the yield between patients with manifest atherosclerosis and patients with risk factors for atherosclerosis only. AB - OBJECTIVE: The purpose of this study was to investigate whether screening for internal carotid artery stenosis (ICAS) and aneurysm of the abdominal aorta (AAA) is indicated in patients with either manifest atherosclerotic disease or with only risk factors for atherosclerosis. STUDY DESIGN: Data were obtained for 2274 patients enrolled in the SMART study, an ongoing single-center, prospective cohort study of patients referred to our vascular center with manifest atherosclerotic disease (peripheral atherosclerotic disease [PAD]; transient ischemic attack [TIA], stroke, or ICAS; AAA; angina pectoris; or myocardial infarction [MI]) or with only risk factors for atherosclerosis (diabetes mellitus, hypertension, hyperlipidemia). The presence of ICAS or AAA was determined with duplex scanning and ultrasonography. RESULTS: The prevalence of ICAS 70% or greater is low in patients with risk factors for atherosclerosis only (1.8%-2.3%), intermediate in patients with angina pectoris or MI (3.1%), and highest in patients with PAD (12.5%) or AAA (8.8%). The prevalence of AAA 3 cm or larger is low in patients with risk factors for atherosclerosis only (0.4-1.6%), intermediate in patients with angina pectoris or MI (2.6%), and highest in patients with PAD (6.5%) or TIA, stroke, or ICAS (6.5%). The prevalence of AAA larger than 5 cm is low in all of the considered patient groups. The yield of screening can be optimized through selection on the basis of simple patient characteristics. In patients with PAD, selecting those with advanced age (>54 years) increased the prevalence of ICAS to 21.8%. Selecting patients with lower diastolic blood pressure (<83 mm Hg) increased the prevalence of ICAS to 17.9%. In patients with both advanced age and lower diastolic blood pressure, the prevalence of ICAS increased to 34.7%. Selecting patients with advanced age increased the prevalence of AAA 3 cm or larger to 9.6%. In patients with TIA, stroke, or ICAS, selecting those with advanced age increased the prevalence of AAA 3 cm or larger to 8.2%. Selecting patients with taller stature (>169 cm) increased the prevalence of AAA 3 cm or larger to 9.3%. In patients with advanced age and taller stature, the prevalence of AAA 3 cm or larger increased to 13.1%. CONCLUSIONS: Screening for ICAS should be limited to patients referred with PAD or AAA, especially those with advanced age or with low diastolic blood pressure. Screening for AAA should be limited to patients referred with PAD or with TIA, stroke, or ICAS, particularly those with advanced age or tall stature. In patients referred with angina pectoris or MI and those referred with only risk factors for atherosclerosis, screening cannot be endorsed. PMID- 12764270 TI - Carotid artery stenting: analysis of data for 105 patients at high risk. AB - OBJECTIVES: Carotid artery stenting (CAS) has been recommended as an alternative to carotid endarterectomy (CEA) by some clinicians. However, recently published clinical trials have reported 30-day stroke and death rates of 10% to 12%. This prompted review of our experience with CAS in patients at high risk, to document our results and guide further use of CAS. METHODS: From September 1996 to the present, we performed 114 consecutive CAS procedures in 105 patients. Sixty-three patients were men (60%) and 42 patients were women (40%), with mean age of 70 years (range, 45-93 years). Indications for CAS included recurrent stenosis after previous CEA in 74 patients (65%), primary lesions in 32 patients at high risk (28%), and carotid stenosis with previous ipsilateral radiation therapy in 8 patients (7%). Asymptomatic stenosis (>80%) was managed in 70 patients (61%), and symptomatic lesions (>50%) were treated in 44 patients (39%). RESULTS: CAS was technically successful in all patients. Mean severity of stenosis before CAS was 87% +/- 6%, compared with 9% +/- 4% after CAS. Two patients (1.9%) died, 1 of reperfusion-intracerebral hemorrhage and 1 of myocardial infarction 10 days after discharge; and 1 patient (0.95%) had a stroke (retinal infarction), for a 30-day stroke and death rate of 2.85%. Two patients (1.9%) had transient neurologic events. No cranial nerve deficits were noted. No neurologic complications have been noted in the last 27 patients (26%). CONCLUSIONS: A 30-day stroke and death rate of 2.85% in our experience demonstrates acceptability of CAS as an alternative to repeat operation or primary CEA in patients at high risk or in patients with radiation-induced stenosis. We recommend further clinical investigation of CAS and participation in clinical trials by vascular surgeons. PMID- 12764271 TI - Improvement in accuracy of diagnosis of carotid artery stenosis with duplex ultrasound scanning with combined use of linear array 7.5 MHz and convex array 3.5 MHz probes: validation versus 489 arteriographic procedures. AB - OBJECTIVE: Validity of a method to improve the accuracy of carotid artery duplex scanning was tested in comparison with arteriography. STUDY DESIGN: In 489 patients who had not previously undergone arteriography, 978 carotid arteries were examined with duplex ultrasound scanning. In method A, a linear array 7.5 MHz transducer with pulsed-wave 4.7 MHz Doppler scanning was used. For the diagnosis and grading of carotid stenosis, peak systolic and end-diastolic velocity of the Doppler waves were recorded. Method B consisted of complete ultrasound imaging and color-flow mapping with a convex array 3.5 MHz transducer with pulsed-wave 2.8 MHz Doppler scanning in all patients who had previously undergone method A. Further velocity measurements were performed at the sites of stenosis. The results of methods A and B were compared with data from neurologic assessment and arteriographic studies. RESULTS: Method B showed significantly higher diagnostic agreement with arteriography than did method A (K 95% confidence interval [CI], 0.87-0.93 vs 0.79-0.85; P <.05), and the number of mistakes in grading stenosis was significantly lower (primarily because of decreased overestimation) in patients with internal carotid kinking (>60 degrees of angulation) (P <.05), distal stenosis (>20 mm from bifurcation) (P <.01), or wide acoustic shadowing (>1 cm) (P <.01) and in those without these conditions (P <.05). Compared with arteriography, diagnostic accuracy with the new method proved higher for carotid stenoses 50% or greater, 60% or greater, 70% or greater, and 80% or greater; no statistically significant difference was found for carotid stenosis 96% or greater or for carotid occlusion. Compared with data from neurologic assessment and arteriography, method B proved more accurate than method A in designating patients for carotid endarterectomy (P =.014). CONCLUSIONS: The new method significantly improved diagnostic reliability of duplex ultrasound scanning, especially in carotid arteries with kinking, distal stenosis, or wide acoustic shadowing (32.2% of all arteries studied). In clinical practice, we suggest additional use of a lower frequency transducer in cases in which these three conditions are found or suspected at first scanning. PMID- 12764272 TI - Computational analysis of effects of external carotid artery flow and occlusion on adverse carotid bifurcation hemodynamics. AB - OBJECTIVE: This is a computational analysis of the effects of external carotid artery (ECA) flow, waveform, and occlusion geometry on two hemodynamic wall parameters associated with intimal hyperplasia and atherosclerosis. Study design Transient three-dimensional fluid mechanics analysis was applied to a standard carotid artery bifurcation. Mean internal carotid artery (ICA) flow was maintained at 236 mL/min with a normal waveform. ECA flow was increased from zero to 151 mL/min (64% of ICA flow) with both a normal biphasic waveform and a damped waveform. Geometry of five ECA occlusions was studied: distal, proximal stump, smooth, smooth without carotid sinus, and optimal reconstruction.Primary outcome measures Two time-averaged and area-averaged hemodynamic wall parameters were computed from the velocity and wall shear stress (WSS) solutions, ie, wall shear stress angle gradient (WSSAG) and oscillatory shear index (OSI). Both local and area-averaged hemodynamic wall parameters were computed for the distal common carotid artery (CCA) and the proximal ICA. RESULTS: When ECA flow with a normal waveform is increased from zero to 151 mL/min, area-averaged WSS values increase in the CCA, from 3.0 to 4.4 dynes/cm(2) (46%), and in the ICA, from 16.5 to 17.1 dynes/cm(2) (4%); minimum local WSS values in the carotid sinus remain less than 1 dyne/cm(2); maximum local values of WSSAG and OSI are observed in the carotid sinus and increase from 3.5 to 9.1 radian/cm (160%) and 0.23 to 0.46 (100%), respectively; CCA plus ICA area-averaged WSSAG increases by 52%, and OSI increases by 144%; and damping of the ECA waveform has little effect on local or area-averaged WSSAG but reduces OSI to 68%. When the ECA is occluded, the minimum local WSS in the carotid sinus is less than 1 dyne/cm(2). However, if the carotid sinus is removed or the CCA-ICA geometry hemodynamically optimized, the minimum WSS is approximately 4 dynes/cm(2). Similarly, eliminating the carotid sinus markedly reduces local maximum WSSAG, from 3.0-3.5 radian/cm to 0.3 radian/cm, and reduces local maximum OSI from 0.22-0.49 to 0.04. Area-averaged WSSAG and OSI over the CCA and ICA are reduced by approximately 50% with elimination of the carotid sinus. CONCLUSIONS: The degree of adverse carotid bifurcation hemodynamics as measured with WSSAG and OSI is directly proportional to ECA flow. The marked difference in normal ICA and ECA flow waveforms does not contribute to adverse wall hemodynamics. Location of an ECA occlusion (distal, proximal, stump, smooth) does not affect adverse carotid hemodynamics; however, marked improvement is obtained with elimination of the carotid sinus. PMID- 12764273 TI - Comparison of treatment plans for peripheral arterial disease made with multi station contrast medium-enhanced magnetic resonance angiography and duplex ultrasound scanning. AB - OBJECTIVE: This study was undertaken to investigate the effects of substituting multi-station total outflow contrast medium-enhanced magnetic resonance angiography (CE-MRA) for color duplex ultrasound (US) scanning on treatment planning in the diagnostic workup of patients with suspected or known peripheral arterial occlusive disease. Patients and methods One hundred consecutive patients referred because of suspected or proved peripheral arterial occlusive disease to a University Hospital underwent both aortoiliac duplex US scanning and multi station total outflow CE-MRA. For 73 of these patients (57% men; mean age, 62 years) treatment or treatment plans could be retraced. Eighteen patients also underwent femoro-popliteal duplex US scanning. Three experienced vascular surgeons retrospectively formulated two sets of treatment plans based on standardized clinical parameters and either duplex US scanning or CE-MRA. The main outcome measure was proportion of patients for whom the treatment plan matched actual treatment without additional use of intra-arterial digital subtraction angiography. Actual treatment, based on all available information, including results of duplex US scanning, CE-MRA, and any other diagnostic tests, served as the standard of reference. RESULTS: Duplex US scanning provided enough information for treatment planning in 46, 45, and 53 patients versus 67, 68, and 66 patients when CE-MRA was used (surgeons 1, 2, and 3, respectively; surgeons 1 and 2, P <.001; surgeon 3, P =.007). Treatment plans based on duplex US scanning exactly matched actual treatment in 37 of 73 patients (51%; surgeon 1), 36 of 73 patients (49%; surgeon 2), and 46 of 73 patients (63%; surgeon 3). Treatment plans based on CE-MRA exactly matched actual treatment in 56 of 73 patients (77%; surgeon 1), 55 of 73 patients (75%; surgeon 2), and 51 of 73 patients (70%; surgeon 3). Positive predictive value and negative predictive value of duplex US scanning as measures of ability to discriminate between surgical and nonsurgical treatment were 0 of 0 (undefined) and 43 of 46 (93%), 1 of 2 (50%) and 40 of 43 (93%), and 5 of 5 (100%) and 44 of 48 (92%) for surgeons 1, 2, and 3, respectively. For CE-MRA, positive and negative predictive values were 11 of 13 (85%) and 50 of 54 (93%), 10 of 12 (83%) and 51 of 56 (91%), and 8 of 13 (62%) and 48 of 53 (91%), respectively, for surgeons 1, 2, and 3. CONCLUSION: Compared with aorto-iliac and femoro-popliteal duplex US scanning, multi-station total outflow CE-MRA is more effective for treatment planning in most patients with known or suspected peripheral arterial occlusive disease. PMID- 12764274 TI - Meta-analysis of polytetrafluoroethylene bypass grafts to infrapopliteal arteries. AB - CONTEXT: Reports of polytetrafluoroethylene (PTFE) bypass grafting to the infrapopliteal arteries have often used survival analysis of acceptable quality to describe a wide range of long-term results. In theory, these results may be combined if variability between series and time intervals is considered. OBJECTIVE: Meta-analysis was performed to gain insight into long-term graft patency and foot preservation after PTFE bypass grafting to infrapopliteal arteries. DATA SOURCE: Studies published from 1982 through 2001 were identified from the PubMed database and pertinent original articles. STUDY SELECTION: Three investigators selected 43 studies that used survival analysis, reported 2-year patency rates, and included at least 15 bypass procedures. Data extraction and transformation: Based on standard life-tables or survivor curves, an interval success rate was calculated for each month in each series. The monthly success rates were combined across series, enabling construction of pooled survivor curves. DATA SYNTHESIS: Random-effects meta-analysis yielded 5-year pooled estimates (SE) of 30.5% (7.6%) for primary graft patency, 39.7% (5.5%) for secondary graft patency, and 55.7% (5.0%) for foot preservation. During the entire follow-up, pooled estimates were slightly higher for series of PTFE grafts with adjunctive procedures compared with series of PTFE grafts only. Sensitivity analysis: A simulation using only unfavorable assumptions showed a decrease of less than 5% at 5 years for all outcomes, and smaller differences at subgroup meta-analysis. Funnel plots suggested that publication bias was unlikely. CONCLUSION: This meta-analysis indicated moderate success for PTFE bypass grafts to infrapopliteal arteries, but the role of adjunctive procedures at the distal anastomosis remains uncertain. PMID- 12764275 TI - Noninvasive ultrasound measurements of aortic intima-media thickness: implications for in vivo study of aortic wall stress. AB - OBJECT: The abdominal aorta (AA) has a predilection for aneurysm formation. An etiologic factor may be underlying aortic wall stress. The purpose of this study was to examine whether the intima-media thickness (IMT) of the AA, as a surrogate to arterial wall thickness, can be measured noninvasively with satisfactory results to calculate circumferential wall stress, and to evaluate regional and gender differences in wall stress. METHODS: Sixty-five middle-aged healthy subjects were examined with B-mode ultrasound to determine the diameter and IMT in the infrarenal AA, common carotid artery (CCA), common femoral artery (CFA), and popliteal artery (PA). Blood pressure was measured noninvasively in the brachial artery. Wall stress was calculated according to the law of LaPlace. RESULTS: Intraobserver variability for the IMT in the AA showed a coefficient of variation of 11%. IMT was thickest in the AA compared with the CCA, CFA, and PA (P <.001). There was a gender difference in IMT in the CFA (P <.05) and PA (P <.01) but not in the AA. Greater wall stress was found in the AA than in the CCA (P <.001) and PA (P <.001), with men having greater wall stress in all studied arterial regions. CONCLUSIONS: Aortic IMT can be satisfactorily studied in vivo with noninvasive B-mode ultrasound. There are gender differences in IMT and wall stress, and the largest wall stress is found in the AA in men, which might be important in aneurysm development. PMID- 12764276 TI - Oscillatory shear stress increases smooth muscle cell proliferation and Akt phosphorylation. AB - PURPOSE: Hemodynamic forces affect smooth muscle cell (SMC) proliferation and migration both in vitro and in vivo. However, the effects of oscillatory shear stress (SS) on SMC proliferation and signal transduction pathways that control survival are not well described. METHODS: Bovine aortic SMC were exposed to arterial levels of oscillatory SS (14 dyne/cm(2)) with an orbital shaker; control cells were exposed to static conditions (0 dyne/cm(2)). Cell number and (3)[H]thymidine incorporation were measured after 1, 3, or 5 days of SS. Activation of the Akt pathway was assessed with the Western blot technique. Specificity of the phosphatidylinositol 3-kinase (PI3K) pathway was determined with the Western blot technique with the inhibitors LY294002 (10 micromol/L) or wortmannin (25 nmol/L). RESULTS: Arterial levels of oscillatory SS increased SMC cell number by 20.1 +/- 3.7% and (3)[H]thymidine incorporation by 33.4% +/- 6.8% at 5 days. To identify whether SS increased activity of the SMC survival pathway, Akt activation was measured. SMC exposed to SS demonstrated increased Akt phosphorylation compared with control cells, with maximal phosphorylation at 60 minutes. Both PI3K inhibitors specifically inhibited the increase in Akt phosphorylation in SMC exposed to oscillatory SS. CONCLUSION: SMC directly respond to oscillatory SS by increasing DNA synthesis, proliferation, and activation of the PI3K-Akt signal transduction pathway. These results suggest a mechanism of SMC survival and proliferation in response to endothelial-denuding arterial injury. PMID- 12764277 TI - Altered proliferative responses of dermal fibroblasts to TGF-beta1 may contribute to chronic venous stasis ulcer. AB - PURPOSE: Venous ulcer fibroblasts demonstrate decreased proliferative responses to growth factor stimulation, suggesting cellular senescence. However, the role of chronic venous insufficiency (CVI) disease progression and extracellular matrix (ECM) proteins in agonist-induced cellular proliferation is ill-defined. We hypothesize that CVI-induced fibroblast proliferative resistance to growth factors worsens with disease progression and is regulated by the composition of ECM. METHODS: Fibroblast explants were isolated from biopsy specimens from two patients without CVI and 16 patients with CVI of the lower calf (LC) and lower thigh (LT) and stratified according to CEAP disease severity: non-CVI (NC; n = 2), class 2-3 (n = 5), class 4 (n = 5), class 5 (n = 3), and class 6 (n = 3). Proliferation experiments were standardized with a neonatal foreskin fibroblast cell line (HS68). A 10-day course and dose response experiment with 0, 0.5, 1.0, 2.5, 5, 10, and 20 ng/mL of transforming growth factor-beta(1) (TGF-beta(1)) demonstrated maximal cell proliferation at 5 ng/mL of TGF-beta(1) on day 4. Under these conditions, CVI dermal fibroblasts were challenged with and without TGF beta(1) and evaluated for proliferative responses on plates coated with polystyrene, collagen, and fibronectin. RESULTS: No differences in unstimulated proliferation were observed in LT and LC fibroblasts from patients with class 2-3 disease and LT fibroblasts from patients with class 4 and 5 disease, compared with NC and HS68 cells. LC fibroblasts from patients with class 4 disease (P <.05) and class 5 disease (P <.001), and LC (P <.001), and LT fibroblasts from patients with class 6 disease (P <.001) proliferated to a lesser degree than did NC and HS68 cells. The diminished proliferation observed in class 4 LC cells was reversible with TGF-beta(1) stimulation (P <.004); however, class 5 and class 6 LC and LT fibroblasts did not respond to stimulation with TGF-beta(1). Collagen increased proliferation of HS68 cells with (P <.05) and without (P <.01) TGF beta(1), compared with cells grown on polystyrene, but did not increase proliferative responses in NC or CVI fibroblasts with and without TGF-beta(1). Similarly, fibronectin increased proliferation of HS68 cells (P <.05) compared with cells grown on polystyrene, but did not alter proliferation in CVI fibroblasts. Fibronectin did seem to inhibit TGF-beta(1)-induced proliferation observed in class 4 LC cells. CONCLUSION: These data indicate that clinical disease progression correlates with cellular dysfunction. Fibroblasts from patients with class 2-3 disease retain their unstimulated and agonist- induced proliferative capacity, compared with NC and HS68 cells. The onset of inflammatory skin changes (class 4 and class 5 disease) diminishes agonist induced proliferation, and ulcer formation (class 6 disease) severely inhibits it. In addition, the composition of ECM does not affect TGF-beta(1)-induced proliferation of fibroblasts in CVI. PMID- 12764278 TI - Sustained inhibition of experimental neointimal hyperplasia with a genetically modified herpes simplex virus. AB - OBJECTIVE: Reported herein is a potential strategy for sustained smooth muscle cell (SMC) inhibition with a virulence-attenuated herpes simplex virus (HSV). Experiments were conducted in vitro to demonstrate selective SMC cytotoxicity and in vivo to demonstrate reduced neointimal hyperplasia (NIH) in a clinically relevant animal model. METHODS: In vitro: Cultured human umbilical artery smooth muscle cells (UASMC) and venous endothelial cells (HUVEC) were exposed to varying multiplicities of infection (MOI) of a gamma(1)34.5-deleted HSV-1 virus (R849). Cell survival was assessed at 48 and 72 hours with a colorimetric MTT viability assay. In vivo: New Zealand White rabbit external jugular veins (n = 21) were exposed to R849 (2.5 x 10(6) pfu/mL) or culture medium at 110 to 120 mm Hg for 10 minutes, then fashioned as vein patches on carotid arteries. Carotid arteries were ligated distally to decrease blood flow and stimulate a hyperplastic response (ultra-low shear stress model). After 2, 4, 12, and 24 weeks, patched segments were perfusion-fixed with glutaraldehyde and morphometrically examined for NIH formation. RESULTS: In vitro: At 48 hours, R849 exhibited preferential cytotoxicity to UASMC compared with HUVEC, with 11% +/- 10% of UASMCs and 49% +/- 8% of HUVECs surviving after infection with MOI = 25 (P <.05). Higher MOI resulted in poor survival of both cell lines. In vivo: Blood flow was similarly reduced in all animals both at surgery (0.9 +/- 0.1 mL/min vs 1.6 +/- 0.3 mL/min) and at harvest (2.7 +/- 0.4 mL/min vs 2.5 +/- 0.5 mL/min). R849-infected patches exhibited markedly less NIH than control patches did at 2 weeks (162 +/- 14 microm vs 49 +/- 6 microm; P <.05), 4 weeks (190 +/- 27 microm vs 67 +/- 8 microm; P <.05), and 12 weeks (233 +/- 18 microm vs 113 +/- 2 microm; P <.05). CONCLUSION: The virulence-attenuated HSV strain R849 demonstrates selective cytotoxicity for SMC and is capable of sustained inhibition of NIH in an experimental model of vein graft failure. PMID- 12764280 TI - Stent-graft treatment of a large internal carotid artery vein graft aneurysm. AB - Aneurysm degeneration of internal carotid artery interposition vein grafts can occur, and their repair can be complicated by repeated neck dissection and difficulty in obtaining distal artery control. We used a covered stent graft to exclude a large internal carotid artery interposition vein graft aneurysm. Retrograde flow was induced in the internal carotid artery so that an embolization protection balloon could be placed through the aneurysm. The stent graft was then placed with embolization protection. The repair was performed without complication, and the aneurysm remained successfully excluded at 6-month follow-up. PMID- 12764279 TI - Paracrine secretion of transforming growth factor-beta1 in aneurysm healing and stabilization with endovascular smooth muscle cell therapy. AB - BACKGROUND: Identification of molecular factors involved in artery wall stabilization after extracellular matrix injury elicited by inflammation and proteolysis has a major role in the development of new therapies for atherosclerosis. A study from our group demonstrated that endovascular seeding of vascular smooth muscle cells (VSMCs) promotes healing and stabilizes experimental aneurysms by downregulating matrix metalloproteinase and upregulating tissue inhibitor of metalloproteinase and collagen gene expression. We analyzed expression of transforming growth factor-beta (TGF-beta) and its receptors in experimental aneurysms treated with endovascular VSMC therapy. METHODS AND RESULTS: Aneurysms were generated in Fischer 344 rats by 14-day orthotopic implantation of a segment of guinea pig abdominal aorta (xenograft). During an endovascular repeat operation, syngeneic VSMCs were seeded in the aneurysm, always resulting in aneurysm diameter stabilization after 8 weeks, whereas diameter of control aneurysms infused with culture medium further increased. Seven days after repeat operation the intima or thrombus was separated from the aneurysmal wall in the two groups. Reverse transcriptase polymerase chain reaction with the domestic gene 18s as a standard demonstrated that aneurysm stabilization was associated with a statistically significant increase in TGF beta(1), but not TGF-beta(2) or TGF-beta(3), messenger RNA levels in the intima. Enzyme-linked immunosorbent assay demonstrated increased TGF-beta(1) protein in the aneurysmal wall. mRNA levels of the two serine and threonine kinase TGF-beta receptors remained unchanged. CONCLUSIONS: Healing and stabilization of aneurysms with endovascular cell therapy is associated with a specific pattern of gene expression, resulting in paracrine secretion of TGF-beta(1). Our study provides insight into the molecular mechanisms of arterial aneurysm healing and stabilization. PMID- 12764281 TI - Double-lumen carotid plaque: a morbid configuration. AB - During analysis of carotid plaque anatomy for a multicenter carotid imaging trial, we examined plaque specimens from 5 patients with double internal carotid artery lumina. Four of the 5 patients had symptoms referable to the lesion. The second lumen was noted when the plaque specimens were examined ex vivo with high resolution (200 microm(3)) magnetic resonance imaging. Plaque structure was correctly identified in only 1 patient preoperatively. However, during retrospective review of the preoperative imaging studies, the second internal carotid artery lumen was identified in 3 patients. PMID- 12764282 TI - Aberrant right subclavian artery syndrome: a case of chronic cough. AB - A young, otherwise healthy man had chronic cough of 16 months' duration. Evaluation revealed an aberrant right subclavian artery. Kommerell's diverticulum without aneurysmal degeneration was present. Imaging studies showed compression of the esophagus but not the trachea. Results of methacholine challenge test were negative for evidence of reactive airway disease, but suggested mild variable intrathoracic obstruction. While aberrant right subclavian artery syndrome most commonly involves dysphagia, our patient's only symptom was cough. Right subclavian artery to right common carotid artery transposition was performed, with oversewing of the subclavian artery stump to the left of the esophagus through a right supraclavicular incision. This treatment was curative, with complete resolution of symptoms. PMID- 12764283 TI - Delayed paraplegia after endovascular repair of abdominal aortic aneurysm. AB - Paraplegia is rare after open repair of infrarenal abdominal aortic aneurysm, and only two cases have been reported after endovascular repair, both due to atheroembolism. Incidence of renal failure after endovascular repair of abdominal aortic aneurysm (EVAR) in patients with normal preoperative renal function is about 8.7%, but is much higher in those with preexisting renal impairment, possibly because of administration of nephrotoxic contrast media during EVAR. We report a case in which contrast medium-induced acute renal failure is believed to have led to delayed paraplegia after EVAR. PMID- 12764284 TI - Surgical treatment of thoracoabdominal aortic mural and floating thrombi extending to infrarenal aorta. AB - The case of a 49-year-old man with thoracoabdominal aortic mural and floating thrombi extending to the infrarenal aorta and occlusion of the common iliac artery is described. He had no factors promoting thrombosis, with a history of thrombectomy of the femoral artery. The thoracoabdominal aortic thrombi were successfully removed with a Forgaty catheter through a thoracotomy under simple aortic clamping and subsequent femoro-femoral cardiopulmonary bypass. Intravascular ultrasound performed through the femoral artery after thrombectomy revealed that little mural thrombi remained and that the celiac, superior mesenteric, and bilateral renal arteries were all patent. PMID- 12764285 TI - Pulseless disease. PMID- 12764286 TI - A new endovascular approach to treatment of acute iliac limb occlusions of bifurcated aortic stent grafts with an exoskeleton. AB - Endovascular aneurysm repair continues to become increasingly popular. As the number of implanted endografts increases, complications will increase as well. We report a new approach to endovascular treatment in two patients with acute iliac limb occlusions of a bifurcated aortic endograft with an endoskeleton. Neither patient required femoral-femoral bypass grafting because of unilateral limb ischemia. We believe this is the optimal primary approach in patients with a bifurcated stent graft with an endoskeleton. PMID- 12764287 TI - Use of axillary artery to maintain cerebral perfusion in a patient requiring reconstruction of the brachiocephalic trunk and coronary arteries: technical note. AB - Our patient had 80% stenosis of the brachiocephalic artery and total occlusion of the left carotid and left subclavian arteries. Ascending aorta to brachiocephalic artery bypass grafting was performed, with a 10 mm Dacron graft. The right axillary artery was cannulated, and during construction of the distal anastomosis cerebral blood flow was from the right axillary artery. We believe this technique may be beneficial in surgery on an artery in which cerebral blood flow depends exclusively. PMID- 12764288 TI - Relevance of federal antitrust statutes to clinical practice. AB - With the advent of Medicare prospective payment systems, health care entities and physicians were forced to decrease expenses by sharing services and to increase revenue by attempting to jointly negotiate better reimbursement from third- party payers. Both activities have raised the specter of prosecution under antitrust laws that have been with us for more than a century but are poorly understood by practicing physicians. Recent monopolistic activities in the health care arena have prompted the Federal Trade Commission to file actions under specific acts of Congress, eg, the Sherman Anti-Trust Act, Clayton Act, Federal Trade Commission Act, Robinson-Patman Act, and Celler-Kefauver Act. Inasmuch as it is likely that specialties such as vascular surgery as a business will undergo substantial transformation, physicians need to be aware of the severe civil and criminal sanctions imposed if they are found guilty; fines that are several times actual damages; activities that raise antitrust issues including utilization review, medical staff privileges for competing specialties, participating provider agreements, and predatory pricing; and affirmative defenses and relief available in terms of specific exemptions. As health care providers react, innovate, and adjust to stay solvent, their business strategies will surely continue to be scrutinized for antitrust behavior by federal and state officials. The physician must have a basic understanding of the groundrules that govern any contemplated business strategy so that common pitfalls may be averted. PMID- 12764289 TI - The ethics of clinical pathways and cost control. PMID- 12764290 TI - Regarding "The Carotid Surgery for Ischemic Stroke trial". PMID- 12764291 TI - Regarding "New method to create a vascular arteriovenous fistula in the arm with an endoscopic technique". PMID- 12764292 TI - Regarding "A prospective study of ultrasound-guided thrombin injection of femoral pseudoaneurysm: a trend toward minimal medication". PMID- 12764301 TI - Minor's rights versus parental rights: review of legal issues in adolescent health care. AB - The right of adolescents to access confidential health care is sensitive and controversial. Recent challenges in the court system to adolescents' right to access abortion and contraception are eroding current law, including the Roe v Wade decision. The prospect of more than a million pregnancies in individuals under the age of 20 years in the United States with increasingly fewer alternatives to pregnancy is concerning. New regulations under the Health Insurance Portability and Accountability Act are adding yet another layer of complexity to the care of adolescents. Understanding legal issues surrounding adolescent rights to care can help the health care provider make appropriate care available to this age group. Keywords previously identified in CINAHL and MEDLINE were used to perform the literature search. LexisNexis was the search engine used to identify the laws and statutes. PMID- 12764300 TI - Developing our future: seeing and expecting the best in youth. PMID- 12764302 TI - Social outcomes of early childbearing: important considerations for the provision of clinical care. AB - Traditional understanding of adolescent childbearing includes a belief that the social and economic consequences for both mother and child are almost universally negative. Recent research, however, reveals that the outcomes resulting from the birth of children to young mothers are diverse and complex. Social circumstances contributing to adolescent pregnancy and parenting, such as poverty, poor educational opportunity, and violence, are also causes of less than optimal outcomes for young mothers and their children; thus, attributions of cause and effect are difficult to make. By reviewing outcomes research that approaches this question in a number of more creative ways, clinicians can gain a more complete understanding of this phenomenon. It is hoped that health care providers with access to new evidence about the social consequences of adolescent childbearing will have improved success and satisfaction in providing competent, compassionate care to pregnant and parenting adolescents. PMID- 12764303 TI - Abnormal uterine bleeding in adolescents. AB - Menarche is an important event during adolescence. For most girls, it marks completion of puberty and the onset of reproductive capability. Menarche usually occurs when both breast and pubic hair development are at Tanner stage 4. Menstrual problems are common during adolescence due to slow maturation of the hypothalamic-pituitary-ovarian axis and can last 2 to 5 years after menarche. Although most problems are explained by anovulation, other causes must be considered and excluded in a logical and cost-effective manner. Frequently, the bleeding problems observed in adolescence require evaluation and intervention. PMID- 12764304 TI - Adolescents' experiences of childbirth: contrasts with adults. AB - Most of what is known about the meaning women assign to the experience of childbirth in the United States is based primarily on studies of Euro American, highly educated, married women of middle to higher income levels. Yet almost half a million adolescents give birth annually. This exploratory, qualitative study was conducted with 25 adolescents in an alternative school, partnered with the juvenile justice system, who had given birth. An open-ended question format was used for the interviews, and analysis was conducted by using extended case methodology. The study revealed a number of differences between the existing literature reports about women's interpretations of birth and the meanings assigned to childbirth experiences by the adolescents in this study. A key difference was the relationship between the pain of childbirth and responsibility for their child. The results provide an entree into understanding unique characteristics of giving birth as an adolescent and potential roles health care providers can play to promote a positive experience. PMID- 12764305 TI - Factors that influence smoking in adolescent girls. AB - A secondary analysis of data from the public-use version of the National Longitudinal Study of Adolescent Health (Add Health) was used for this research study. Variables and constructs that significantly influence smoking in adolescent girls are identified. These variables and constructs include residential mother-daughter connectedness, self-concept, residential mother's smoking behavior, and friends' smoking behavior. All variables and constructs identified are significantly associated with smoking in white adolescent girls. Residential mother-daughter connectedness and friends' smoking behavior are significantly associated with smoking in black adolescent girls. Friends' smoking behavior is the only variable in this study found to be significantly associated with smoking in Hispanic adolescent girls. Results of this secondary analysis suggest that the influential and protective factors related to smoking vary with race/ethnicity. PMID- 12764306 TI - Barriers to physical activity perceived by adolescent girls. AB - Girls in the United States are at high risk for inactivity and resultant overweight or obesity. Perceived barriers to physical activity are critical factors for health care professionals to consider when intervening with this population to increase physical activity. This descriptive study identified barriers to physical activity reported by girls in middle school. Participants were recruited from two middle schools in the Midwest. Using a Likert-type scale, a total of 77 ethnically diverse girls, ages 11 to 14, responded to 23 items representing barriers to physical activity participation. Mean scores and percentages were computed for each barrier statement. The top barriers to physical activity that emerged for the girls in this investigation were "I am self-conscious about my looks when I exercise" and "I am not motivated to be active." Strategies that health professionals can use to counsel girls of this age in overcoming barriers to physical activity are presented. This health related information can enhance anticipatory guidance to girls. PMID- 12764307 TI - Adolescent sexual risk assessment. AB - This article describes the use of a self-administered event history calendar and interviews about sexual partners, sexual activity, and contraception within the broader context of the adolescent's life. Constant comparative analysis was used to analyze the event history calendar interview data. Detailed 5- to 9-year sexual histories were obtained on the event history calendars, including patterns of sexual activity progression and triggers for unintended sexual intercourse. Discussions of protective and risky sexual decision making were stimulated by adolescents' self-report of sexual risk behavior in their own words and reflective responses on the event history calendar. Both the adolescents and the interviewers thought the calendars encouraged recall and discussion of sexual risk behavior. PMID- 12764309 TI - Adolescents' experiences of dating and intimate partner violence: "once is not enough". AB - Pregnant and parenting adolescents represent a significant proportion of the individuals directly affected by intimate partner violence. Although screening tools are useful, it is important to ask very specific questions to open lines of communication with adolescents. This article presents a clinical case that highlights the challenges of screening adolescents for intimate partner violence and offers suggestions for health care providers. Specific examples of screening questions are offered. PMID- 12764308 TI - Centering pregnancy: a model for pregnant adolescents. AB - Recent exploration of the experiences of pregnant and parenting adolescents has uncovered the need to address the unique developmental, social, and cultural aspects of adolescent pregnancy. Many adolescents, especially those from urban areas, live in communities with limited opportunities, poverty, violence, and a lack of support. Programs that help young women discover their inner strength, create environments for empowerment, and build community may help adolescents to achieve goals and parent successfully. Centering Pregnancy is a model of group prenatal care that provides for the assessment, education, and support of pregnant women and may be particularly useful in adolescent populations. The model is described and the ways adolescents may benefit from Centering Pregnancy's unique design is discussed. PMID- 12764310 TI - Weight gain during pregnancy. PMID- 12764311 TI - Guidelines for adolescent preventive services (GAPS). PMID- 12764321 TI - Are we asking and are we listening? PMID- 12764330 TI - Homicide and suicide risks associated with firearms in the home: a national case control study. AB - STUDY OBJECTIVE: I test the hypothesis that having a gun in the home is a risk factor for adults to be killed (homicide) or to commit suicide. METHODS: Two case control analyses were based on national samples of subjects 18 years of age or older. Homicide and suicide case subjects were drawn from the 1993 National Mortality Followback Survey. Living control subjects were drawn from the 1994 National Health Interview Survey. Ten control subjects matched by sex, race, and age group were sought for each case subject. RESULTS: The homicide sample consisted of 1,720 case subjects and 8,084 control subjects. Compared with adults in homes with no guns, the adjusted odds ratio (OR) for homicide was 1.41 (95% confidence interval [CI] 1.20 to 1.65) for adults with a gun at home and was particularly high among women (adjusted OR 2.72; 95% CI 1.89 to 3.90) compared with men (adjusted OR 1.23; 95% CI 1.01 to 1.49) and among nonwhite subjects (adjusted OR 1.74; 95% CI 1.37 to 2.21) compared with white subjects (adjusted OR 1.27; 95% CI 1.03 to 1.56). Further analyses revealed that a gun in the home was a risk factor for homicide by firearm means (adjusted OR 1.72; 95% CI 1.40 to 2.12) but not by nonfirearm means (OR 0.83; 95% CI 0.62 to 1.11). The suicide sample consisted of 1,959 case subjects and 13,535 control subjects. The adjusted OR for suicide was 3.44 (95% CI 3.06 to 3.86) for persons with a gun at home. However, further analysis revealed that having a firearm in the home was a risk factor for suicide by firearm (adjusted OR 16.89; 95% CI 13.26 to 21.52) but was inversely associated with suicide by other means (adjusted OR 0.68; 95% CI 0.55 to 0.84). CONCLUSION: Having a gun at home is a risk factor for adults to be shot fatally (gun homicide) or commit suicide with a firearm. Physicians should continue to discuss with patients the implications of keeping guns at home. Additional studies are warranted to address study limitations and to better understand the implications of firearm ownership. PMID- 12764331 TI - Factors associated with severity of motorcycle injuries among young adult riders. AB - STUDY OBJECTIVE: A cohort of 4,729 junior college students in Taiwan was studied to determine risk factors for increased severity of motorcycle injuries. METHODS: Crash characteristics were collected by using self-administered questionnaires soon after a student was involved in a motorcycle crash. The proportional odds model with generalized estimating equations, with correlated ordinal responses for collisions categorized into not injured, mildly injured, and severely injured categories, was used to determine the odds of injury severity. RESULTS: A total of 1,889 motorcycle crashes involving 1,284 persons occurred over a 20-month period from November 1994 to June 1996. There were 1,339 noninjuries, 474 mild injuries, and 76 severe injuries. The adjusted odds ratio (OR) of rural to urban roads having a greater level of injury severity was 1.64. Compared with noncollisions, collisions with a moving car (adjusted OR=1.76), a parked car (adjusted OR 1.90), or another stationary object (adjusted OR=2.31) increased the odds for a greater level of injury severity. Riders using Sanyang (adjusted OR=1.63) and Yamaha (adjusted OR=1.39) motorcycles had greater odds of being involved in a crash with a greater level of injury compared with those riding Kymco motorcycles. Darkness (adjusted OR=1.65) and greater speeds (adjusted OR=1.63 to 4.69) also increased the odds of greater injury severity. CONCLUSION: At the time of motorcycle crashes, factors such as being on rural roads, collisions with a heavier object, some motorcycle makes, darkness, and greater speeds increased the severity of motorcycle injuries among these young adult riders. PMID- 12764332 TI - Motorcycle casualties sustained during Daytona Beach Bike Week 2000: lessons learned. AB - STUDY OBJECTIVE: In March 2000, an estimated 500,000 people attended an annual motorcycle rally in Daytona Beach, FL, where approximately 64,000 residents live year-round. The media reported 15 deaths during this 10-day event. To more comprehensively assess the extent of trauma and need for emergency medical care, we investigated all motorcycle crashes, regardless of outcome. METHODS: Motorcycle-related crash data from local medical examiner, hospital, emergency medical services (EMS), and police sources were linked. Frequencies of crashes, injuries, hospitalizations, and deaths were determined, and EMS use data were analyzed. RESULTS: During Bike Week 2000, 570 people were involved in 281 motorcycle-related crashes. Two hundred thirty (40%) people were injured, of which 147 (64%) sought treatment in emergency departments, 72 (31%) were hospitalized, and 11 (5%) died. In crashes between motorcycles and passenger cars, individuals exposed as motorcycle occupants were 8.7 times more likely to be injured than car occupants (95% confidence limit 1.7, 15.7). Of 205 EMS dispatches for motorcycle-related crashes, two thirds resulted in transport to an ED. Data needed to assess known risk factors (eg, alcohol use, speed, lack of helmet use) were not routinely ascertained at either the crash site or ED. CONCLUSION: Although fatalities first called attention to the problem, nonfatal injuries outnumbered deaths 20:1. The manpower resources of civil service and health resources could become overwhelmed or exhausted in circumstances in which many people are injured or killed throughout a relatively long period. The situation deserves future study. Better risk factor surveillance is needed to help prevent crashes. PMID- 12764333 TI - National Highway Traffic Safety Administration (NHTSA) notes. Trauma system agenda for the future. PMID- 12764334 TI - Commentary: Trauma systems: a key factor in homeland preparedness. PMID- 12764336 TI - Treatment for alcohol and other drug problems: Closing the gap. PMID- 12764335 TI - Assessing substance abuse treatment need: a statewide hospital emergency department study. AB - STUDY OBJECTIVE: Health care providers in hospital emergency departments rarely take substance abuse histories or assess associated treatment need. This study compares documentation of psychoactive drug-related diagnoses for adult ED patients in medical records with treatment need assessed through self-report, toxicologic screening, and Diagnostic and Statistical Manual of Mental Disorders, 4th edition (DSM-IV), criteria. METHODS: A statewide, 2-stage, probability sample survey was conducted in 7 Tennessee general hospital EDs from June 1996 to January 1997. Main outcome measures were the prevalence of diagnosed substance abuse problems, positive bioassay results, denied use, and treatment need. Sensitivity and multivariate analyses were conducted by using varied case definitions of treatment need. RESULTS: Thirty-one percent (95% confidence interval [CI] 27.3% to 34.7%) of screened ED patients (n=1,330) had positive test results for substance use. Their prevalence of denial of use in the 30 days before the survey ranged from 10% for alcohol (95% CI 5.7% to 14.3%) to 100% for phencyclidine. One percent of all ED patients (n=1,502) had a recorded diagnosis of substance abuse. By contrast, as many as 27% (95% CI 23.3% to 31.8%) were assessed as needing substance abuse treatment on the basis of a comprehensive case definition that accounted for denial and positive test results. A sensitivity analysis using other case definitions is also presented. For example, 4% (95% CI 2.8% to 5.3%) of patients met the very strict definition of DSM-IV current drug dependence only. Under the comprehensive case definition, TennCare patients (adjusted odds ratio [OR] 1.63; 95% CI 1.30 to 2.05) and Medicare patients (adjusted OR 2.50; 95% CI 1.34 to 4.65) showed excess treatment need relative to the privately insured. Excess need was also exhibited by patients reporting 1 or more prior ED visits in the past year (adjusted OR 1.62; 95% CI 1.13 to 2.31) and by patients taking 2 or more hours to reach the ED after the onset of injury or illness (adjusted OR 1.54; 95% CI 1.16 to 2.04). Treatment need was inversely associated with age. Irrespective of case definition, less than 10% of ED patients who needed substance abuse treatment were receiving such treatment. CONCLUSION: EDs can be important venues for detecting persons in need of substance abuse treatment. PMID- 12764337 TI - Cyclobenzaprine with ibuprofen versus ibuprofen alone in acute myofascial strain: a randomized, double-blind clinical trial. AB - STUDY OBJECTIVE: We evaluate the analgesic and side effects of adding cyclobenzaprine to ibuprofen in emergency department patients with acute myofascial strain. METHODS: A randomized, prospective, double-blind study was conducted at an urban teaching ED with an annual census of 44,000. One hundred two patients aged 18 to 70 years with acute myofascial strain caused by minor trauma within the prior 48 hours were included, and 77 patients completed the protocol. Each patient received a single dose of 800 mg of ibuprofen in the ED and a vial of 6 capsules containing 800 mg of ibuprofen to take every 8 hours as needed after discharge from the ED. In addition, 51 patients received a single dose of 10 mg of cyclobenzaprine and a vial of 6 capsules containing 10 mg of cyclobenzaprine to take every 8 hours as needed after discharge from the ED; the remaining 51 patients received an identically labeled placebo capsule and vial of placebo capsules to take every 8 hours as needed after discharge from the ED. Patients rated the intensity of their pain on a 100-mm visual analog scale (VAS) at baseline; 30, 60, 90, 120, and 180 minutes; and 24 and 48 hours after treatment. Telephone follow-up was obtained at 24 and 48 hours, and side effects were elicited at 24 and 48 hours by means of open-ended questioning. RESULTS: The patients in each group were similar with regard to diagnosis and baseline pain score. The number of patients who did not complete the protocol and the number of those who required additional analgesia were similar in both groups. Over the 48 hours of the protocol, the mean VAS score for the combination group decreased from 60.4 to 35.6, and the mean VAS score for the ibuprofen alone group decreased from 62.2 to 35.4. The mean VAS scores between groups across time was not statistically significant (P =.962, repeated-measures analysis of variance). At both 24 and 48 hours, central nervous system side effects were reported more frequently in the patients receiving cyclobenzaprine (16 [42%] versus 7 [18%] at 24 hours and 15 [39%] versus 5 [13%] at 48 hours, respectively). CONCLUSION: In ED patients with acute myofascial strain, the addition of cyclobenzaprine to ibuprofen does not improve analgesia but is associated with a greater prevalence of central nervous system side effects. PMID- 12764338 TI - A prospective, randomized, double-blind comparison of buffered versus plain tetracaine in reducing the pain of topical ophthalmic anesthesia. AB - STUDY OBJECTIVE: We determine whether buffering ocular tetracaine hydrochloride reduces the pain of instillation. METHODS: We conducted a prospective, randomized, double-blind, 2-treatment, 2-period crossover, single center study of healthy volunteers 18 years of age or older. Participants were randomized to receive either 2 drops of buffered or plain tetracaine in a randomly assigned eye. After a mean wash out period of 24 days (range 7 to 54 days), participants returned to have 2 drops of the other medication instilled in the same eye. The participants recorded the pain of instillation on a 100-mm visual analog scale (VAS) immediately and 5 minutes after instillation. Adverse events were also recorded at these intervals. The primary outcome measure was the intensity of pain as measured on a VAS immediately after instillation. RESULTS: Sixty persons were enrolled in the study, with 100% follow up. Immediately after instillation, the adjusted mean VAS score for buffered tetracaine was 29.1 mm, and the adjusted mean VAS score for plain tetracaine was 16.0 mm. The estimated difference was 13.1 mm (95% confidence interval 6.9 to 19.3 mm). CONCLUSION: Buffering of tetracaine hydrochloride significantly increases the pain of its instillation in healthy volunteers, suggesting that pain with instillation of ocular anesthetics is not dependent on low pH. PMID- 12764339 TI - Interrater reliability in pupillary measurement. AB - STUDY OBJECTIVE: A previous report documented ranges of normal pupil size on the basis of measurements by the principal investigator. In this report, we examine interrater reliability of pupillary measurement. METHODS: According to a randomized double-blind assignment, healthy volunteers received phenylephrine in one eye and sterile water in the other. After a wait period, the principal investigator and then other observers, using a gauge with a modified Haab scale, took measurements of each pupil, both in fluorescent light (2,700 to 5,400 lux) and bright light (>54,000 lux), whereas the contralateral pupil was concealed. For the first pupil observed in each session, each observer also provided a gestalt judgment of whether the pupil was dilated. In the study's main comparison, paired differences between principal investigator and nonprincipal investigator measurements were summarized by using the median and interquartile range for measurements taken in both light intensities. Interrater agreement in diagnosing pupillary dilation was also calculated for gestalt judgment and for simple measurement. RESULTS: Among 149 principal investigator/nonprincipal investigator pairs taken from 102 participants, principal investigator room-light measurements were 0.2 mm (median 0.2 mm; interquartile range -0.4 to 0.7 mm) greater than those from other observers, but there was no difference in bright light measurements (median 0 mm; interquartile range -0.5 to 0.4 mm). Nevertheless, principal investigator/nonprincipal investigator percentage agreement in judging dilation by means of measurement was high in both light intensities (85% to 86%). In pupils with gestalt judgment of the presence or absence of pupillary dilation, percentage agreement between nonprincipal investigator observers was higher for measurement (75% to 82%) than for gestalt judgment (61%). CONCLUSION: There was no systematic difference between principal investigator and nonprincipal investigator bright-light measurements, supporting the range of normal values published previously. Interrater agreement in diagnosing pupillary dilation by means of measurement was high. PMID- 12764341 TI - Intravenous sodium valproate versus prochlorperazine for the emergency department treatment of acute migraine headaches: a prospective, randomized, double-blind trial. AB - STUDY OBJECTIVE: We compare the efficacy of intravenous sodium valproate versus prochlorperazine for the emergency department treatment of acute migraine headache. METHODS: We performed a randomized, prospective, double-blind trial performed at a tertiary care military ED. Forty patients, aged 18 to 65 years, presenting with typical migraine symptoms were enrolled. Patients were randomized to receive either 10 mg of prochlorperazine or 500 mg of valproate intravenously over 2 minutes. Pain, nausea, and sedation were assessed by using a standard visual analog scale (VAS). Changes in VAS scores were compared between groups from baseline to end point by using a rank sum test, over time by using 2-way repeated-measures analysis of variance, and by requirement for rescue at 60 minutes by using the Fisher exact test. RESULTS: Comparison of the change in median VAS scores over 60 minutes revealed that sodium valproate was significantly less effective than prochlorperazine in reducing pain or nausea (P <.001). Median improvements in VAS pain scores (binomial confidence intervals) were as follows: 64.5 mm (48.1 to 75.6 mm) for prochlorperazine versus 9 mm (-3 to 39.6 mm) for sodium valproate. Median improvements in VAS nausea scores were as follows: 35.5 mm (13.2 to 47.9 mm) for prochlorperazine versus 2 mm (-1.3 to 11 mm) for sodium valproate. There was no significant difference (P =.603) detected in the median changes in VAS scores for sedation: -4 mm (-29.9 to 8.6 mm) for prochlorperazine versus 0 mm (-6.6 to 6 mm) for sodium valproate. Comparison of the mean VAS time curves for pain and nausea also demonstrated a significant difference (both P <.001) but not for sedation (P =.232). In post hoc analysis, valproate failed to elicit significant improvement in pain or nausea scores over time, whereas prochlorperazine improved pain by 30 minutes (P <.001) and nausea by 15 minutes (P =.002). At the conclusion of the study, 15 (79%) of 19 patients receiving valproate required rescue treatment compared with 5 (25%) of 20 patients receiving prochlorperazine (P <.001). CONCLUSION: Prochlorperazine was statistically and clinically superior to sodium valproate for the treatment of the pain and nausea associated with acute migraine headaches. PMID- 12764340 TI - Beyond tissue plasminogen activator: mechanical intervention in acute stroke. AB - Mechanical interventions in acute ischemic stroke promise to provide emergency physicians with tools to treat patients in whom conventional thrombolysis might be ineffective or contraindicated, including most patients with stroke who arrive at the emergency department beyond the 3-hour time window for intravenous tissue plasminogen activator. A systematic MEDLINE literature review was performed. Endovascular interventions currently in early human clinical trials include the use of lasers, ultrasonography, angioplasty, microsnares, and a variety of clot retrieval devices. Potential advantages of these approaches include more rapid recanalization of occluded vessels, reduced or no exposure to fibrinolytic agents, and a longer treatment window. Early safety trials are promising, with serial improvements in device design to minimize trauma to cerebrovascular endothelium and accelerate vessel recanalization. The purpose of this review is to provide the emergency medicine community with an understanding of these promising and emerging approaches to acute stroke therapy. PMID- 12764342 TI - Fracture of the lateral process of the talus associated with snowboarding. AB - Snowboarding is one of the fastest-growing winter sports and is associated with a relatively high rate of ankle injuries. Presented is a patient who, after falling while snowboarding, complained of lateral ankle pain and was misdiagnosed with an ankle sprain. Further workup revealed a lateral process of the talus fracture, an injury that is rare outside of snowboarding. A lateral process of the talus fracture should be suspected when there is a history of inversion with dorsiflexion and there is tenderness over the lateral process of the talus. Results of plain films are negative up to 40% of the time, and therefore a computed tomographic scan is the imaging modality of choice. Treatment includes immobilization and not bearing weight for 4 to 6 weeks for nondisplaced fractures or open reduction and fixation for displaced fractures. Up to two thirds of patients with lateral process of the talus fractures report chronic pain. Early recognition may decrease this relatively high rate of morbidity. PMID- 12764343 TI - Interexaminer agreement on the American Board of Emergency Medicine oral certification examination. AB - STUDY OBJECTIVE: One of the principal criticisms of performance assessments, particularly those using an oral format to evaluate physician performance, is the lack of interrater reliability. The objective of this study was to assess interexaminer agreement in the scoring of candidates taking the American Board of Emergency Medicine (ABEM) oral certification examination. METHODS: Examiner observer pairs independently scored individual candidates on each of the 6 simulated cases selected for the October 1999 examination. Simple proportionate agreement for "acceptable" and "unacceptable" scores on critical actions and performance ratings were used to measure interexaminer (examiner-observer) agreement. Further analyses were performed to assess interexaminer agreement at the level of the raw scores of all performance ratings. The candidate's actual score was determined entirely by the examiner administering the case. The study examiner was present only to observe and independently score the candidate. Examiners were blinded to each other's scoring. All examiner-observer, examiner candidate, and observer-candidate pairings were uniquely randomized and occurred only once during the examination. RESULTS: Of 564 candidates, 186 were randomly selected for scoring by examiner-observer pairs drawn from a pool of 107 trained oral examiners. Interexaminer agreement for 47 critical actions was 97% (95% confidence interval [CI] 96% to 98%), and interexaminer agreement for 68 performance ratings was 95% (95% CI 94% to 96%). Further analyses of raw scores of performance ratings indicated that 94% (95% CI 93% to 95%) of 2,648 pairs of ratings differed by 1 point or less. CONCLUSION: High interexaminer agreement on the scoring of the ABEM oral certification examination supports the reliability of this final step in the certification process. On the basis of these findings, ABEM has incorporated this methodology into the administration of each oral examination as an ongoing quality control measure. PMID- 12764344 TI - Disaster medicine and the emergency medicine resident. PMID- 12764345 TI - Keeping lists and naming names: habitual patient files for suspected nontherapeutic drug-seeking patients. AB - Emergency departments commonly keep files of patients who are suspected of frequently visiting them and fabricating symptoms to obtain prescription drugs, usually opioids, for nontherapeutic purposes. Such files have previously been given names such as "frequent flyer file," "repeater log," "kook-book," "problem patient file," "patient alert list," or "special needs file." Little has been written about the ethical, legal, and regulatory considerations that should be taken into account when establishing, maintaining, and using such files. This article explores these issues. The term "habitual patient files" is proposed because it is descriptive without being judgmental. PMID- 12764348 TI - High-dose therapy in multiple myeloma. PMID- 12764349 TI - Histiocytic disorders. PMID- 12764350 TI - Antiphospholipid antibodies and thrombosis: strength of association. AB - Antiphospholipid antibodies, that is, lupus anticoagulants and anticardiolipin antibodies, are associated with thrombosis and obstetric complications in the antiphospholipid syndrome. Venous thrombosis occurs mostly in the lower limbs, with or without pulmonary embolism, and cerebral ischemia and transient ischemic attacks are the most common arterial events. Overall, the prevalence of thrombosis is about 30%, the rate of first event approximates 1%/year, and that of recurrence of patients not receiving anticoagulation is about 10-29%/year. The presence of lupus anticoagulants carries an odds ratio for thrombosis ranging from 5 to 16, and that of anticardiolipin antibodies from nonsignificant to 18. The detection of anti-beta2-glycoprotein I, but not antiprothrombin, antibodies might also help to identify antiphospholipid-positive patients at risk of thrombosis. Unfractionated or low-molecular-weight heparin followed by oral anticoagulation represents the current treatment of both arterial and venous thrombosis. However, uncertainty still exists about the optimal duration and intensity of oral anticoagulation following the first event. Several therapeutic clinical trials are currently being conducted, which soon clarify these issues. The prevalence of obstetric complications is about 15-20%. The presence of lupus anticoagulants carries an odds ratio for recurrent miscarriages and fetal death ranging from 3.0 to 4.8, whereas that of anticardiolipin antibodies goes from 0.86 to 20. Unfractionated or low-molecular-weight heparin in combination with low-dose aspirin represents the current standard of treatment of pregnant antiphospholipid-positive women to prevent recurrent obstetric complications. Upon treatment, the live birth rate increases from 0-40% to 70-80%. PMID- 12764351 TI - Effect of bcl-2 antisense oligodeoxynucleotides on drug sensitivity of leukemic cells. AB - INTRODUCTION: We investigated the effect of two antisense oligodeoxynucleotides, previously selected with the help of computer-aided RNA structure prediction, on drug sensitivity, bcl-2 expression and apoptosis of leukemia cells. The drugs tested were etoposide (VP-16), cytarabine (Ara-C), daunorubicin (DNR) and arsenic trioxide (As(2)O(3)). MATERIALS AND METHODS: The experimental assays were performed with cultures, IC(50) of leukemic cells to drugs, immunochemistry and flow cytometry. RESULTS: The results showed that the two antisense oligodeoxynucleotides significantly reduced IC(50) levels for VP-16, Ara-c, DNA and As(2)O(3), inhibited bcl-2 gene expression and induced apoptosis of leukemic cells. CONCLUSIONS: Computational prediction of antisense efficacy is faster than other methods and more cost-efficient. This could hasten the development of sequences for both research and clinical applications. PMID- 12764352 TI - Long-term outcomes of polycythemia vera patients treated with pipobroman as initial therapy. AB - From 1968 to 1993, 179 newly diagnosed patients with polycythemia vera (PV) were enrolled in a prospective study using pipobroman as first chemotherapy. Among them, 140 fulfilled the Polycythemia Vera Study Group criteria for PV, and 39 patients (22%) can be considered as idiopathic erythrocytosis (IE). Vascular events occurred in 10% of IE and 20% of PV patients and solid tumors in 7.7% of IE and 12.8% of PV patients. There were no differences between PV and IE patients with regard to progression to myelofibrosis (MF), leukemic events and overall survival. Overall, 98.3% of patients initially responded to pipobroman, with very mild toxicity. A total of 164 PV patients who received more than 1 year of pipobroman were analyzed for long-term evolution. The actuarial risk of thrombosis was 15.6 and 23.8% at 10 and 18 years, respectively. In all, 21 patients developed a solid tumor during follow-up, added and/or switched drugs being a risk factor. Actuarial risk of MF was as low as 4.9 and 9.4% at 10 and 15 years, respectively. Actuarial risk of leukemia was 14.4 and 18.7% at 10 and 15 years, respectively. Hyperleukocytosis at diagnosis was the only variable significantly associated with higher risk of leukemia. The median survival was 15.5 years, with two initial adverse prognostic factors: age above 60 years and hyperleukocytosis. Despite an increasing risk of leukemia with time, survival was not lower when compared to the French matched population. Only age and hyperleukocytosis at diagnosis were found to have a prognostic value in PV. PMID- 12764353 TI - Bone marrow transplantation for patients with acquired severe aplastic anemia using cyclophosphamide and antithymocyte globulin: the experience from a single center. AB - Between 1998 and 2001, 31 (24 male, 7 female) patients with severe aplastic anemia (SAA) and a median age of 19 years (range, 4-39 years) received an allogeneic bone marrow transplantation. Marrow donors were genotypically HLA identical siblings in 30 cases and a monozygous twin in one case. The median time from diagnosis to bone marrow transplantation was 1 month (range, 0.5-5 months). Conditioning regimen consisted of cyclophosphamide (CY) combined with antithymocyte globulin (ATG), in all patients. For graft-versus-host disease (GvHD) prophylaxis, all patients received methotrexate and cyclosporin. A total of 84% of patients had sustained grafts, whereas 16% rejected grafts between 3 and 20 months after transplantation. Of the five rejecting patients, three are alive with successful second engraftments and two died from infections. Acute grade II-IV GvHD was seen in only 11% of patients. A limited chronic GvHD was seen in one patient. With a median follow-up of 18 months (range, 5-42 months), survival rate was 86% and Karnofsky score was at least 90%. This study confirms the high success rate of the CY/ATG regimen in SAA allografted from an HLA identical sibling. Early and late graft failure remains a problem and may require modification of this regimen. PMID- 12764354 TI - Therapeutic concordance of two portable monitors and two routine automatic oral anticoagulant monitoring systems using as reference the manual prothrombin time technique. AB - Two models of capillary blood prothrombin time (PT) monitoring systems were evaluated for analytical performance and then compared with two routine PT systems using the reference manual technique and a high-sensitivity thromboplastin. Two sets of 60 and 80 plasmas were analyzed from anticoagulated patients stabilized over 3 months in an INR range 2-3.5 for therapy. Capillary PT determination was performed in two portable monitors, CoaguChek S and CoaguChek PT (Roche Diagnostics), and plasma automatic methods were Neoplastine/STA (Diagnostics Stago) and PT-FibrinogenHsPlus/ACL7000 (Instrumental Laboratories). Thromboplastin Bilbao (TBi), an in-house high-sensitivity rabbit thromboplastin (ISI=1.08), recommended as the reference reagent by an External Spanish Oral Anticoagulant Quality Assessment, was used in the PT manual technique. The two monitors' coefficients of correlation with the reference system were 0.74 for CoaguChek S and 0.81 for CoaguChek PT. The automatic routine systems showed a correlation of 0.92 (Neoplastine/STA) and 0.91 (PT-FbHsPlus/ACL7000). Clinical agreement expressed as the percentage of simple correlation ranged between 75.0% (CoaguChek S) and 88.9% (Neoplastine/STA). The systems having the best kappa index with the manual technique were CoaguChek PT (71.9%) and the Neoplastine/STA system (73%). The routine PT management systems exhibited better correlation and percentage of concordance when using the TBi/manual technique than did the portable monitors, which moreover performed unequally in this regard. PMID- 12764355 TI - Complete remission induced by G-CSF in a patient with acute myeloid leukemia with t(8;21)(q22;q22). AB - We describe a case of acute myeloid leukemia (AML) with t(8;21) in which complete remission (CR) was obtained with G-CSF given at 10 microg/kg in the absence of concomitant cytotoxic chemotherapy. CR was achieved following 2 weeks of therapy and confirmed by investigating minimal residual disease by four-color flow cytometry analysis. During treatment with G-CSF, maturing cells with cytoplasmic Auer Rods were observed in the peripheral blood, suggesting a differentiation effect. This case adds further evidence for a specific role of G-CSF in the treatment of AML with t(8;21), namely in patients who are not eligible for aggressive chemotherapy. PMID- 12764356 TI - Acute tumor lysis syndrome in large B-cell non-Hodgkin lymphoma induced by steroids and anti-CD 20. PMID- 12764357 TI - Controlled growth of tetrapod-branched inorganic nanocrystals. AB - Nanoscale materials are currently being exploited as active components in a wide range of technological applications in various fields, such as composite materials, chemical sensing, biomedicine, optoelectronics and nanoelectronics. Colloidal nanocrystals are promising candidates in these fields, due to their ease of fabrication and processibility. Even more applications and new functional materials might emerge if nanocrystals could be synthesized in shapes of higher complexity than the ones produced by current methods (spheres, rods, discs). Here, we demonstrate that polytypism, or the existence of two or more crystal structures in different domains of the same crystal, coupled with the manipulation of surface energy at the nanoscale, can be exploited to produce branched inorganic nanostructures controllably. For the case of CdTe, we designed a high yield, reproducible synthesis of soluble, tetrapod-shaped nanocrystals through which we can independently control the width and length of the four arms. PMID- 12764358 TI - Dextran templating for the synthesis of metallic and metal oxide sponges. AB - Silver or gold-containing porous frameworks have been used extensively in catalysis, electrochemistry, heat dissipation and biofiltration. These materials are often prepared by thermal reduction of metal-ion-impregnated porous insoluble supports (such as alumina and pumice), and have surface areas of about 1 m(2) g( 1), which is typically higher than that obtained for pure metal powders or foils prepared electrolytically or by infiltration and thermal decomposition of insoluble cellulose supports. Starch gels have been used in association with zeolite nanoparticles to produce porous inorganic materials with structural hierarchy, but the use of soft sacrificial templates in the synthesis of metallic sponges has not been investigated. Here we demonstrate that self-supporting macroporous frameworks of silver, gold and copper oxide, as well as composites of silver/copper oxide or silver/titania can be routinely prepared by heating metal salt-containing pastes of the polysaccharide, dextran, to temperatures between 500 and 900 degrees C. Magnetic sponges were similarly prepared by replacing the metal salt precursor with preformed iron oxide (magnetite) nanoparticles. The use of dextran as a sacrificial template for the fabrication of metallic and metal oxide sponges should have significant benefits over existing technologies because the method is facile, inexpensive, environmentally benign, and amenable to scale up and processing. PMID- 12764359 TI - Nanostructured artificial nacre. AB - Finding a synthetic pathway to artificial analogs of nacre and bones represents a fundamental milestone in the development of composite materials. The ordered brick-and-mortar arrangement of organic and inorganic layers is believed to be the most essential strength- and toughness-determining structural feature of nacre. It has also been found that the ionic crosslinking of tightly folded macromolecules is equally important. Here, we demonstrate that both structural features can be reproduced by sequential deposition of polyelectrolytes and clays. This simple process results in a nanoscale version of nacre with alternating organic and inorganic layers. The macromolecular folding effect reveals itself in the unique saw-tooth pattern of differential stretching curves attributed to the gradual breakage of ionic crosslinks in polyelectrolyte chains. The tensile strength of the prepared multilayers approached that of nacre, whereas their ultimate Young modulus was similar to that of lamellar bones. Structural and functional resemblance makes clay- polyelectrolyte multilayers a close replica of natural biocomposites. Their nanoscale nature enables elucidation of molecular processes occurring under stress. PMID- 12764360 TI - Inorganic materials: nanocrystals branch out. PMID- 12764361 TI - In BCR-ABL-positive cells, STAT-5 tyrosine-phosphorylation integrates signals induced by imatinib mesylate and Ara-C. AB - In BCR-ABL-positive cells, the transcription factor STAT-5 is constitutively activated by tyrosine phosphorylation. STAT-5 activation results in upregulation of bcl-X(L) and increased resistance to induction of apoptosis. Here, we investigated the effects of imatinib mesylate and cytosine arabinoside (Ara-C) on STAT-5 tyrosine-phosphorylation, cellular proliferation and induction of apoptosis in cell lines and primary hematopoietic cells. Imatinib mesylate treatment strongly suppressed STAT-5 tyrosine-phosphorylation in K562 and primary CML blasts. In contrast to JAK-2 and PI-3-kinase inhibition, exposure of K562 cells to imatinib mesylate resulted in obvious suppression of proliferation. Reduced cell growth was due to specific induction of caspase activation followed by apoptotic cell death. In addition, we investigated the effects of Ara-C on STAT-5 tyrosine-phosphorylation. Exposure to Ara-C resulted in significant downregulation of STAT-5 tyrosine-phosphorylation and inhibition of DNA binding. Treatment of K562 cells with Ara-C in combination with imatinib mesylate revealed synergistic effects at the level of STAT-5 tyrosine-phosphorylation and DNA binding, Hck tyrosine-phosphorylation, cell growth and induction of apoptosis. Overall, in this report we demonstrate that STAT-5 tyrosine-phosphorylation is a specific target of imatinib mesylate and Ara-C. Our results suggest that, in combination therapy, inhibition of STAT-5 tyrosine-phosphorylation may be responsible for synergistic or additive effects on BCR-ABL-positive cells. PMID- 12764362 TI - Current CML therapy: progress and dilemma. AB - Rarely has progress in treatment of leukemia been as dramatic and convincing as with the BCR-ABL tyrosine kinase inhibitor imatinib.(1) Imatinib induces remissions of CML as fast as hydroxyurea, achieves rates of cytogenetic remissions that by far exceed those induced by interferon alpha and has a toxicity profile as favourable as that of hydroxyurea and much superior to that of interferon alpha.(2) In addition, the causal approach of this new drug, which may well serve as a model for new treatment modalities in other neoplasias is reassuring. PMID- 12764364 TI - Donor leukocyte infusions in acute myelogenous leukemia. PMID- 12764363 TI - Detection of minimal residual disease in hematologic malignancies by real-time quantitative PCR: principles, approaches, and laboratory aspects. AB - Detection of minimal residual disease (MRD) has prognostic value in many hematologic malignancies, including acute lymphoblastic leukemia, acute myeloid leukemia, chronic myeloid leukemia, non-Hodgkin's lymphoma, and multiple myeloma. Quantitative MRD data can be obtained with real-time quantitative PCR (RQ-PCR) analysis of immunoglobulin and T-cell receptor gene rearrangements, breakpoint fusion regions of chromosome aberrations, fusion-gene transcripts, aberrant genes, or aberrantly expressed genes, their application being dependent on the type of disease. RQ-PCR analysis can be performed with SYBR Green I, hydrolysis (TaqMan) probes, or hybridization (LightCycler) probes, as detection system in several RQ-PCR instruments. Dependent on the type of MRD-PCR target, different types of oligonucleotides can be used for specific detection, such as an allele specific oligonucleotide (ASO) probe, an ASO forward primer, an ASO reverse primer, or germline probe and primers. To assess the quantity and quality of the RNA/DNA, one or more control genes must be included. Finally, the interpretation of RQ-PCR MRD data needs standardized criteria and reporting of MRD data needs international uniformity. Several European networks have now been established and common guidelines for data analysis and for reporting of MRD data are being developed. These networks also include standardization of technology as well as regular quality control rounds, both being essential for the introduction of RQ PCR-based MRD detection in multicenter clinical treatment protocols. PMID- 12764365 TI - Prevention and therapy of fungal infections in bone marrow transplantation. PMID- 12764366 TI - Stem cells and diversity. PMID- 12764367 TI - Diffuse alveolar hemorrhage: an evolving problem? PMID- 12764368 TI - Incomplete DJH rearrangements as a novel tumor target for minimal residual disease quantitation in multiple myeloma using real-time PCR. AB - The hypervariable regions of immunoglobulin heavy-chain (IgH) rearrangements provide a specific tumor marker in multiple myeloma (MM). Recently, real-time PCR assays have been developed in order to quantify the number of tumor cells after treatment. However, these strategies are hampered by the presence of somatic hypermutation (SH) in VDJH rearrangements from multiple myeloma (MM) patients, which causes mismatches between primers and/or probes and the target, leading to a nonaccurate quantification of tumor cells. Our group has recently described a 60% incidence of incomplete DJH rearrangements in MM patients, with no or very low rates of SH. In this study, we compare the efficiency of a real-time PCR approach for the analysis of both complete and incomplete IgH rearrangements in eight MM patients using only three JH consensus probes. We were able to design an allele-specific oligonucleotide for both the complete and incomplete rearrangement in all patients. DJH rearrangements fulfilled the criteria of effectiveness for real-time PCR in all samples (ie no unspecific amplification, detection of less than 10 tumor cells within 10(5) polyclonal background and correlation coefficients of standard curves higher than 0.98). By contrast, only three out of eight VDJH rearrangements fulfilled these criteria. Further analyses showed that the remaining five VDJH rearrangements carried three or more somatic mutations in the probe and primer sites, leading to a dramatic decrease in the melting temperature. These results support the use of incomplete DJH rearrangements instead of complete somatically mutated VDJH rearrangements for investigation of minimal residual disease in multiple myeloma. PMID- 12764369 TI - Requirement for the PI3K/Akt pathway in MEK1-mediated growth and prevention of apoptosis: identification of an Achilles heel in leukemia. AB - The Raf/MEK/ERK kinase cascade plays a critical role in transducing growth signals from activated cell surface receptors. Using DeltaMEK1:ER, a conditionally active form of MEK1 which responds to either beta-estradiol or the estrogen receptor antagonist 4 hydroxy-tamoxifen (4HT), we previously documented the ability of this dual specificity protein kinase to abrogate the cytokine dependency of human (TF-1) and murine (FDC-P1 and FL5.12) hematopoietic cells lines. Here we demonstrate the ability of DeltaMEK1:ER to activate the phosphatidylinositol 3-kinase (PI3K)/Akt/p70 ribosomal S6 kinase (p70(S6K)) pathway and the importance of this pathway in MEK1-mediated prevention of apoptosis. MEK1-responsive cells can be maintained long term in the presence of beta-estradiol, 4HT or IL-3. Removal of hormone led to the rapid cessation of cell proliferation and the induction of apoptosis in a manner similar to cytokine deprivation of the parental cells. Stimulation of DeltaMEK1:ER by 4HT resulted in ERK, PI3K, Akt and p70(S6K) activation. Treatment with PI3K, Akt and p70(S6K) inhibitors prevented MEK-responsive growth. Furthermore, the apoptotic effects of PI3K/Akt/p70(S6K) inhibitors could be enhanced by cotreatment with MEK inhibitors. Use of a PI3K inhibitor and a constitutively active form of Akt, [DeltaAkt(Myr(+))], indicated that activation of PI3K was necessary for MEK1 responsive growth and survival as activation of Akt alone was unable to compensate for the loss of PI3K activity. Cells transduced by MEK or MEK+Akt displayed different sensitivities to signal transduction inhibitors, which targeted these pathways. These results indicate a requirement for the activation of the PI3K pathway during MEK-mediated transformation of certain hematopoietic cells. These experiments provide important clues as to why the identification of mutant signaling pathways may be the Achilles heel of leukemic cell growth. Leukemia treatment targeting multiple signal transduction pathways may be more efficacious than therapy aimed at inhibiting a single pathway. PMID- 12764370 TI - Expression of CD1d by myelomonocytic leukemias provides a target for cytotoxic NKT cells. AB - Natural killer T (NKT) cells with an invariant T-cell receptor for alpha galactosylceramide (alphaGalCer) that is presented by CD1d have been reported to be cytotoxic for myelomonocytic leukemia cells. However, the necessity for leukemia cell CD1d expression, the role of alphaGalCer, and the cytotoxic mechanisms have not been fully elucidated. We evaluated these issues with myeloid leukemia cells from 14 patients and purified NKT cells that were alphaGalCer/CD1d reactive. CD1d was expressed by 80-100% of cells in three of seven acute myeloid leukemias (AMLs) and by 28-77% of cells in five of six juvenile myelomonocytic leukemias (JMML). CD1d+ AML cells were myelomonocytic or monoblastic types, and CD1d+ JMML cells were differentiated and CD34-. Cytotoxicity required leukemia cell CD1d expression and was increased by alphaGalCer (P<0.0001) and inhibited by anti-CD1d mAb (P<0.001). The perforin/granzyme-B pathway of NKT cells caused up to 85% of cytotoxicity, and TNF-alpha, FASL, and TRAIL mediated additional killing. CD56+ NKT cells expressed greater perforin and were more cytotoxic than CD56 NKT cells without alphaGalCer (P<0.0001), but both subpopulations were highly and equally cytotoxic in the presence of alphaGalCer. We conclude that CD1d expression is stage-specific for myelomonocytic leukemias and could provide a target for NKT-cell-mediated immunotherapy. PMID- 12764371 TI - Timed-sequential chemotherapy with concomitant granulocyte colony-stimulating factor for newly diagnosed de novo acute myelogenous leukemia. AB - EMA, consisting of etoposide, mitoxantrone, and cytarabine, is a timed-sequential chemotherapy (TSC) regimen and an efficacious option for induction treatment of acute myelogenous leukemia (AML). Hematopoietic growth factors (HGFs) have been shown to recruit leukemic blasts into cell cycle. We postulated the addition of granulocyte colony-stimulating factor (G-CSF) to EMA (EMA-G) might enhance treatment efficacy. EMA-G consisted of mitoxantrone on days 1-3, cytarabine on days 1-3 and 8-10, etoposide on days 8-10, and G-CSF from day 4 until absolute neutrophil count (ANC) >500/microl. In total, 28 patients were enrolled. All patients had newly diagnosed de novo AML. The median age was 42 years. Of the 27 patients with cytogenetic analysis, six had favorable karyotype, 18 intermediate karyotype, and three unfavorable karyotype. The median follow-up was 37.5 months. The median time for both ANC recovery and last platelet transfusion was 26 days. The toxicities associated with this regimen were no more than those expected with the standard chemotherapy. In all, 24 (86%) patients achieved complete remission (CR), three (11%) patients had no response, and one patient died within 24 h of induction therapy before response could be evaluated. Of the 24 patients who achieved CR, 22 received high-dose cytosine arabinoside and two received allogeneic bone marrow transplant as initial postremission therapy. For the whole cohort, the estimated 3-year survival rate was 67%. The median relapse-free survival was 30.5 months. We conclude that EMA-G regimen is a safe regimen and administration of G-CSF during and after induction treatment is not associated with prolongation of marrow aplasia or acceleration of leukemia relapse. It is efficacious for induction therapy for newly diagnosed de novo AML. A high CR rate can be achieved with only one course of this chemotherapy. PMID- 12764372 TI - Treatment of elderly patients (> or =60 years) with newly diagnosed acute promyelocytic leukemia. Results of the Italian multicenter group GIMEMA with ATRA and idarubicin (AIDA) protocols. AB - In all, 134 elderly patients (median age 66 years, range 60-75 years) with newly diagnosed acute promyelocytic leukemia (APL) were enrolled in two successive protocols of the Italian multicenter group GIMEMA. All patients received an identical induction with all-trans retinoic acid and idarubicin; 116 (86%) entered complete remission (CR), two (2%) were resistant and 16 (12%) died during induction. After CR, 106 patients received further therapy whereas 10 did not, because of refusal (n=5) or toxicity (n=5). Consolidation consisted of three chemotherapy courses in the AIDA protocol (AIDA, 67 patients) or, since 1997, of an amended protocol including only the first cycle (amended AIDA, aAIDA, 39 patients). In the AIDA group, 43 patients (64%) completed consolidation, while seven (11%) and 17 (25%) patients were withdrawn after first and second courses, respectively; nine patients (13%) died in CR and 12 (18%) relapsed. In the aAIDA group, all patients received the assigned treatment; two patients (5%) died in CR and six (15%) relapsed. In the AIDA and aAIDA series, the 3-year overall and discase-free survival rates were 81 and 83% (P=NS), 73 and 72% (P=NS), respectively. We highlight here the frequency and severity of complications linked to intensive chemotherapy in this clinical setting and suggest that, in APL of the elderly, less intensive postremission therapy allows significant reduction of severe treatment-related toxicity and may be equally effective. PMID- 12764373 TI - High frequency of pro-B acute lymphoblastic leukemia in adults with secondary leukemia with 11q23 abnormalities. AB - To evaluate the frequency and cytogenetic and immunophenotypic features of therapy-related, precursor B-cell acute lymphoblastic leukemia (ALL), 152 cases of immature B-cell ALL were reviewed. These were compared to the frequency of therapy-related acute myeloid leukemia (t-AML) during the same time period. Eight ALL cases with a prior diagnosis of malignancy were identified, including six (4.0%) with prior therapy considered to be therapy-related ALL (t-ALL). The t-ALL cases followed treatment for breast carcinoma (two cases), lung carcinoma (two cases), lymphocyte predominance Hodgkin's disease and follicular lymphoma with a latency period of 13 months to 8 years. All t-ALL cases had a pro-B (CD10 negative) immunophenotype with significantly higher expression of CD15 and CD65, compared to the de novo CD10-positive ALL cases. All six t-ALL cases had MLL abnormalities by fluorescence in situ hybridization, and four showed t(4;11)(q21;q23). These represented half of all 11q23-positive adult ALL cases. During the same time period, 4.9% of all AML cases were considered t-AML. There was a 16.7% frequency of 11q23 abnormalities in the t-AML group. Despite the similar frequency in therapy-related disease among ALL and AML cases, there were differences in the frequency of the diseases and t-ALL represented 12% of all therapy-related leukemias. However, t-ALL represented 46% of all 11q23-positive therapy-related leukemias. The immunogenetic features of t-ALL appear distinct and may aid in identifying more cases of this disease type in the future. PMID- 12764374 TI - Substance P--a potent risk factor in childhood lymphoblastic leukaemia. AB - The study focused on determining the expression of substance P (SP) in neoplastic bone marrow cells in childhood acute lymphoblastic leukaemia (ALL) in terms of its mRNA and the level of protein production. An attempt has also been made to demonstrate a correlation of SP with leukaemia risk factors and treatment failure. The study group comprised 120 children treated for ALL. Expression of SP was examined by in situ hybridisation with a 5'-biotinylated probe and by immunocytochemistry with specific anti-human SP antibody. Out of 80 patients with common ALL, the expression of SP was demonstrated in 33 cases (41.2%). In the group of 24 children with pre-B ALL, the presence of SP was noted in six cases (25.0%). Of 16 patients with T-cell leukaemia, SP expression was demonstrated in 13 cases (81.2%). The percentage of immunopositive cells in the SP-positive cases ranged from 79.8 to 97.3. Treatment failure in the children with ALL was closely related to the expression of SP observed at the beginning of treatment. The results showed a connection between the presence of SP-positive blasts and leukaemia relapse. This may indicate that SP expression, involved in the proliferation of the tumour cells, may represent a novel risk factor in ALL. PMID- 12764375 TI - Outcome in patients with nonleukemic granulocytic sarcoma treated with chemotherapy with or without radiotherapy. AB - Granulocytic sarcoma (GS) is an extramedullary tumor composed of immature myeloid cells. The objectives of this study were to describe the frequency, presenting characteristics, and survival in patients with nonleukemic GS by conducting a review of all untreated patients presenting to the MD Anderson Cancer Center between January 1990 and June 2002. In all, 21 patients with nonleukemic GS, 1520 patients with acute myeloid leukemia (AML), and 402 patients with high-risk myelodysplastic syndrome (MDS) were identified. GS occurred in 1.4% of patients with AML, and 1.1% of patients with AML or high-risk MDSs. The median patient age was 57 years (range, 7-81). Among 20 patients with available cytogenetics in tissue and/or bone marrow, six had chromosome 8 abnormalities. The median follow up of surviving patients is 12 months (range, 7-75). In all, 20 patients were treated. Patients were treated with AML-type chemotherapy (n=16), chemotherapy and radiotherapy (n=3), or radiotherapy alone (n=1). A total of 13 patients (65%) achieved complete remission and one patient (5%) achieved partial remission. The median overall survival was 20 months (range, 1-75), median overall failure-free survival was 12 months (range, 1-75). The median survival of patients with chromosome 8 abnormalities was 12 months compared with 40 months of those without (P=0.17). Novel therapies for patients with GS are required. PMID- 12764376 TI - Fludarabine-induced apoptosis of B chronic lymphocytic leukemia cells includes early cleavage of p27kip1 by caspases. AB - B-cell chronic lymphocytic leukemia (B-CLL) is characterized by the accumulation of growth arrested clonal B lymphocytes that undergo apoptosis when treated with fludarabine. To further explore the mechanism for the cell cycle arrest, we examined the expression and activity of cyclin-dependent kinases and inhibitors in primary B-CLL cells. We observed high levels of p27kip1, cyclin D2, cyclin E, cdk2, and cdk4 expression in freshly isolated B-CLL cells. Despite high levels of cyclins and cdks, little cdk2 or cdk4 activity was observed with p27kip1 in complex with cyclinD2/cdk4 and cyclin E/cdk2. Remarkably, when B-CLL cells were treated in vitro with fludarabine, p27kip1 underwent caspase-specific degradation accompanied by an increase in cdk4 activity. We conclude that the G0/G1 arrest of B-CLL cells may protect against apoptosis and that the decrease in p27kip1 expression by caspase cleavage may be a key step in chemotherapy-induced apoptosis in B-CLL. PMID- 12764377 TI - Identification of a novel fusion gene, TTL, fused to ETV6 in acute lymphoblastic leukemia with t(12;13)(p13;q14), and its implication in leukemogenesis. AB - ETS variant gene 6 (ETV6)/translocation, ETS, leukemia (TEL)-involving chromosomal translocations are frequently observed in various hematologic neoplasms. We describe here a novel ETV6-involving translocation, t(12;13)(p13;q14), found in the case of acute lymphoblastic leukemia, in which ETV6 fused with a previously unknown gene, named Twelve-thirteen Translocation Leukemia gene (TTL), at 13q14. TTL was weakly but ubiquitously expressed in normal human tissues as detected by reverse transcribed-PCR. Three TTL splicing forms were identified, TTL-T from a human testis cDNA library, with an open reading frame of 402 bp encoding 133 amino acids (aa), and TTL-B1 and -B2 from a human brain cDNA library. These proteins have no homology to known proteins. In leukemic cells from the patient, both reciprocal fusion transcripts, ETV6/TTL and TTL/ETV6, were expressed. The predominant fusion transcript, TTL/ETV6-1, encodes a predicted 530 aa fusion protein containing 89 aa of the N-terminal TTL fusing to the helix-loop-helix domain and ETS-binding domain of ETV6. Although the function of TTL is yet to be elucidated, our findings will provide another insight into the molecular pathogenesis of leukemia having ETV6-involving translocations. PMID- 12764378 TI - PAX5/ETV6 fusion defines cytogenetic entity dic(9;12)(p13;p13). AB - Recurrent chromosomal abnormalities present in malignant cells often define subentities with unique biological and clinical features. The molecular identification of genes involved in genetic alterations has led to the characterization of fusion genes with neoplastic properties. However, for many nonrandom translocations including the dic(9;12)(p11-13;p11-12), the molecular equivalent has not as yet been identified. The dicentric translocation dic(9;12) is a recurrent chromosome abnormality that accounts for close to 1% of childhood acute lymphoblastic leukemia (ALL). This specific alteration occurs almost exclusively in B-progenitor ALL, and unlike many other nonrandom translocations, is associated with an excellent prognosis. In this work, we provide strong evidence that the PAX5/ETV6 fusion transcript defines the clinical and biological entity that is associated with the presence of a dic(9;12) chromosome. As the PAX5 and ETV6 genes are localized at 9p13 and 12p13, respectively, the cytogenetic description of the dic(9;12)-PAX5/ETV6 rearrangement should be refined to dic(9;12)(p13;p13). PMID- 12764379 TI - Patterns of BCR/ABL gene rearrangements by interphase fluorescence in situ hybridization (FISH) in BCR/ABL+ leukemias: incidence and underlying genetic abnormalities. AB - Interphase fluorescence in situ hybridization (iFISH) is increasingly used for the identification of BCR/ABL gene rearrangements in chronic myeloid leukemia (CML) and acute lymphoblastic leukemia (ALL). In the present study, we have explored the incidence of both typical and atypical iFISH patterns of BCR/ABL gene rearrangements in a series of 168 consecutive BCR/ABL+ patients--135 CML, 31 precursor B-ALL and two acute myeloblastic leukemia (AML) cases--and established their underlying genetic alterations through further molecular and chromosome analyses. Two different FISH probes (Vysis Inc., Downers Grove, IL, USA) were used: the LSI BCR/ABL dual color extra signal (ES) and the dual color dual fusion BCR/ABL probe (D-FISH). Our results show that most BCR/ABL+ patients (83%, including 88% of all CML, 61% of ALL and one of two AML) displayed typical iFISH patterns of either Major (M) BCR/ABL (87% of CML, 13% of ALL and one of the two AML) or minor (m) BCR/ABL gene rearrangements (1% of all CML and 48% of ALL cases) with the two probes. Further molecular and cytogenetic studies confirmed the presence of such typical rearrangements in all except one of these ALL cases who had coexistence of an MBCR/ABL and an mBCR/ABL gene rearrangement together with monosomy 9. In the remaining 29 cases (17%), up to five different atypical iFISH patterns were detected with the ES probe. Atypical iFISH patterns were most frequently due to additional numerical changes--most often supernumerary Philadelphia (Ph) chromosome (7%) but also gain or loss of chromosome 9 (1%) or 22 (1%). Deletion of 9q sequences proximal to the breakpoint were also frequently observed with the ES probe (8%). Application of the D-FISH probe showed that in most of these latter cases (5%) deletion of 22q sequences distal to the breakpoint also occurred. The remaining cases with atypical iFISH had cryptic insertion of BCR in 9q34 (1%). Exact interpretation of each iFISH pattern was supported by FISH on metaphases and molecular determination of the BCR breakpoint. In summary, our results indicate that despite the high incidence of typical iFISH patterns of BCR/ABL gene rearrangements, atypical patterns are also found in BCR/ABL+ acute leukemias; the precise definition of the alteration present in individual cases is dependent on metaphase studies and molecular definition of the breakpoint. PMID- 12764380 TI - Monitoring of minimal residual disease (MRD) by real-time quantitative reverse transcription PCR (RQ-RT-PCR) in childhood acute myeloid leukemia with AML1/ETO rearrangement. AB - The fusion transcript AML1/ETO corresponding to translocation t(8;21)(q22;q22) can be found in approximately 7-12% of childhood de novo AML. Despite the favorable prognosis, some of these patients relapse. Most of MRD studies so far were performed on adults treated not uniformly. Therefore, we analyzed the follow up of 15 AML1/ETO-positive children using real-time quantitative reverse transcription PCR (RQ-RT-PCR), all enrolled in the multicenter therapy trial AML BFM 98. AML1/ETO copy numbers were normalized to the control gene ABL and the results were expressed in copy numbers AML1/ETO per 10 000 copies ABL. At diagnosis, a median of 10 789 copies AML1/ETO was found. A linear decrease to about 10 copies (2-4 log) could be seen in most of the children by the start of consolidation. In the majority of cases they remained positive at this low level during the ongoing therapy. Four children relapsed and two of them had a decrease of less than 2 log before starting consolidation. Three of the relapsed children showed, prior to relapse, an increase of the AML1/ETO fusion transcript at 6, 9, and 11 weeks, respectively. These results suggest that monitoring of minimal residual disease using RQ-RT-PCR could be helpful in detecting patients with a higher risk of relapse. PMID- 12764381 TI - Functional characterization of human CD34+ cells that express low or high levels of the membrane antigen CD111 (nectin 1). AB - Nectins are recently described adhesion molecules that are widely expressed on many tissues, including the hematopoietic tissue. Nectin 1 (CD111) is expressed on a higher proportion of mobilized peripheral blood (mPB) than cord blood (CB) CD34+ cells, and of CD34+/CD38+ cells when compared with CD34+/CD38- cells. We studied functional properties of human CB and mPB CD34+ cells that express low or high levels of CD111. CD34+/CD111(dim) cells contain a higher proportion of cells in G0/G1 phase than CD34+/CD111(bright) cells. CD34+/CD111(bright) cells contain more erythroid progenitors: CFU-E, than their counterparts, which on the opposite contain more HPP-CFC. Limiting dilution analyses demonstrate a higher frequency of immature progenitors: cobblestone-area colony-forming cells, CD34+/CD111(dim) than in CD34+/CD111(bright) cells. In vitro differentiation assays demonstrate a higher frequency of B-, T- and dendritic-cell precursors, but less NK-cell precursors in CD34+/CD111(dim) cells. Evaluation of engraftment in NOD-SCID mice shows that SCID repopulating cells are more frequent among mPB CD34+/CD111(dim) cells. Liquid culture of CD34+/CD111(dim) cells with erythropoietin shows that CD111 expression increases simultaneously with CD36, following CD71 and before glycophorin A expression. In conclusion, immature human hematopoietic progenitors express low levels of CD111 on their surface. During erythroid differentiation CD34+ cells acquire higher levels of the CD111 antigen. PMID- 12764382 TI - Lack of telomerase activity in human mesenchymal stem cells. AB - Telomerase activity and telomere maintenance have been associated with immortality in tumor and embryonic stem cells. Whereas most normal somatic cells are telomerase negative, low levels of this enzyme have been found in adult stem cells from the skin, gut and the hematopoietic system. Here, we show that telomerase activity is not detectable in human mesenchymal stem cells (hMSCs), which have the phenotype SH2+, SH3+, SH4+, CD29+, CD44+, CD14-, CD34- and CD45-, and have the capacity to differentiate into adipocytes, chondrocytes and osteoblasts. These data suggest that hMSCs have a different telomere biology compared to other adult stem cells. Alternatively, true mesenchymal stem cells might be a very rare subpopulation that have a detection level that is below the sensitivity of the TRAP assay. PMID- 12764383 TI - Proinflammatory cytokines and their role in the development of major transplant related complications in the early phase after allogeneic bone marrow transplantation. AB - Serum levels of interleukin-6 (IL-6), interleukin-8 (IL-8) and tumor necrosis factor (TNF)-alpha were frequently measured during the first 30 days after allogeneic bone marrow transplantation (BMT) in 84 consecutive adult patients. Major transplant-related complications (MTCs) occurred in 33% of cases and included veno-occlusive liver disease, idiopathic pneumonia syndrome, severe endothelial leakage syndrome and >grade II acute graft-versus-host disease. Compared with patients having minor complications, those with MTCs developed higher levels at times of maximal clinical signs (all cytokines, P<0.001), between days 0-5 post-BMT (IL-6 and IL-8, P<0.05) and days 6-10 (L-6, P<0.001; IL 8 and TNF, P<0.01) post-BMT. We could not discriminate patterns of cytokine release that were specific for any subtype of MTC. Higher levels of IL-8 during days 0-5 were associated (P=0.044) with early (<40 days) death. Multivariate analysis including patient and transplant characteristics as well as post-BMT levels of C-reactive protein showed that high average levels of one or more of the cytokines within the first 10 days post-BMT were independently associated with MTC (Odd's ratio: 2.3 [1.2-4.5], P=0.011). This study shows that systemic release of proinflammatory cytokines contributes to the development of MTC and provides a rationale for pre-emptive anti-inflammatory treatment in selected patients. PMID- 12764384 TI - Homeobox gene expression profile in human hematopoietic multipotent stem cells and T-cell progenitors: implications for human T-cell development. AB - Class I homeobox (HOX) genes comprise a large family of transcription factors that have been implicated in normal and malignant hematopoiesis. However, data on their expression or function during T-cell development is limited. Using degenerated RT-PCR and Affymetrix microarray analysis, we analyzed the expression pattern of this gene family in human multipotent stem cells from fetal liver (FL) and adult bone marrow (ABM), and in T-cell progenitors from child thymus. We show that FL and ABM stem cells are similar in terms of HOX gene expression, but significant differences were observed between these two cell types and child thymocytes. As the most immature thymocytes are derived from immigrated FL and ABM stem cells, this indicates a drastic change in HOX gene expression upon entry into the thymus. Further analysis of HOX-A7, HOX-A9, HOX-A10, and HOX-A11 expression with specific RT-PCR in all thymocyte differentiation stages showed a sequential loss of 3' region HOX-A cluster genes during intrathymic T-cell development and an unexpected expression of HOX-A11, previously not recognized to play a role in hematopoiesis. Also HOX-B3 and HOX-C4 were expressed throughout thymocyte development. Overall, these data provide novel evidence for an important role of certain HOX genes in human T-cell development. PMID- 12764385 TI - Costimulatory signals distinctively affect CD20- and B-cell-antigen-receptor mediated apoptosis in Burkitt's lymphoma/leukemia cells. AB - CD20 is a B-cell differentiation antigen and known to induce apoptosis in Burkitt's lymphoma/leukemia (BL) cells upon antibody-mediated crosslinking. We examined the biological effect of CD20 crosslinking on BL cell lines and observed that apoptosis induction is accompanied by activation of multiple caspases, including caspase-8, -9, -3, -2, and -7. Further investigation revealed a clear synergism between apoptosis mediated by CD20 and by B-cell antigen receptor (BCR). Examination of the effect of simultaneous crosslinking of other cell surface molecules with crosslinking of CD20 or BCR on apoptosis induction showed that these molecules had either a synergistic or inhibitory effect on induction of apoptosis. It is worth noting that some molecules had a different effect on CD20- and BCR-mediated apoptosis. Simultaneous crosslinking of the molecules CD10, CD22, CD72, and CD80 inhibited BCR-mediated apoptosis, but enhanced CD20 mediated apoptosis. Further studies revealed that regulation of CD20-induced apoptosis by other costimulatory molecules is achieved by modification of caspase activation. CD20-mediated apoptosis in BL cells may provide not only a model for understanding the mechanism regulating clonal selection of B cells but a new therapeutic strategy for BL patients. PMID- 12764387 TI - Myelodysplasia--therapeutic response to novel therapy and the need for new diagnostic groups. PMID- 12764386 TI - Bone marrow stromal-derived soluble factors and direct cell contact contribute to de novo drug resistance of myeloma cells by distinct mechanisms. AB - The tumor microenvironment plays a critical role in determining the fate of tumor cells. We have previously reported that adhesion of human myeloma and leukemia cell lines to the extracellular matrix protein, fibronectin, confers a multidrug resistant phenotype. Mechanisms associated with this cell adhesion-mediated drug resistance are drug-type specific. In the present study, we examined the influence of bone marrow stromal cells (BMSCs) on myeloma cell response to the topoisomerase II inhibitor, mitoxantrone. Apoptosis was inhibited by more than 50% when cells were adhered to BMSCs as compared to myeloma cells maintained in suspension. To investigate the mechanisms contributing to the resistance of myeloma cells in contact with BMSCs, we examined the protective effects of BMSCs under four separate conditions: (1) direct cell contact; (2) BMSCs conditioned medium; (3) medium conditioned by coculturing myeloma cells in direct contact with BMSCs; and (4) medium conditioned by coculturing myeloma cells and BMSCs without direct physical contact. Conditioned medium from BMSCs alone was not sufficient to protect myeloma cells from drug-induced apoptosis; however, soluble factors produced during the myeloma-BMSCs interaction decreased the sensitivity of myeloma cells to mitoxantrone, suggesting a dynamic interaction between myeloma cells and BMSCs. We also found that myeloma cells in direct contact with BMSCs underwent growth arrest, whereas soluble factors produced by myeloma cells BMSCs coincubation stimulated the proliferation of myeloma cells. These data show that both cell-cell adhesion of BMSCs with myeloma cells and soluble factors induced by this cell-cell interaction are involved in the protection of myeloma cells from mitoxantrone-induced apoptosis; however, the mechanisms contributing to the drug resistance are different. PMID- 12764388 TI - Imatinib mesylate (Gleevec(TM)) reduces phlebotomy requirements in polycythemia vera. PMID- 12764389 TI - Rare coincidence of hypertriploid chromosome number and aberrant coexpression of the lymphoid-associated antigen CD5 in acute myeloid Leukaemia FAB M0. PMID- 12764390 TI - Effect of NF-kappaB/Rel inhibition on spontaneous vs chemotherapy-induced apoptosis in AML and normal cord blood CD34+ cells. PMID- 12764391 TI - Frequencies of the major subgroups of precursor B-cell acute lymphoblastic leukemia in Indian children differ from the West. PMID- 12764393 TI - Retention and defective assembly of the B-cell receptor in the endoplasmic reticulum of chronic lymphocytic leukaemia B cells cannot be reverted upon CD40 ligand stimulation. PMID- 12764392 TI - A novel real-time RT-PCR assay for quantification of OTT-MAL fusion transcript reliable for diagnosis of t(1;22) and minimal residual disease (MRD) detection. PMID- 12764394 TI - Questioning the aim of CML therapy in the era of Imatinib? PMID- 12764395 TI - Cytogenetic response to thalidomide treatment in three patients with myelodysplastic syndrome. PMID- 12764396 TI - Retinoic acid and steroid in acute promyelocytic leukemia. PMID- 12764397 TI - The Antihypertensive and Lipid-Lowering Treatment to Prevent Heart Attack Trial (ALLHAT): ALL predictable, and no big surprise out of a HAT? PMID- 12764398 TI - ALLHAT: a saga of missed opportunities. PMID- 12764399 TI - Cardiovascular diseases and G-protein beta3 subunit gene (GNB3) in the era of genomewide scans. PMID- 12764400 TI - Trends in coronary artery disease and associated risk factors in sub-Saharan Africans. AB - It has been suggested that the prevalence of coronary artery disease (CAD) is steadily increasing in sub-Saharan Africa. To address this issue, we conducted a Medline search of English language articles on cardiovascular diseases-and specifically CAD in Africa- from 1966 to 1997. The prevalence of CAD and related complications is relatively low in most regions in Africa compared to that obtained in the economically developed countries, although the situation is rapidly changing due to trends in urbanization, changes in lifestyle, acquisition of technology and the increasing numbers of tertiary care institutions. There are variations in reported prevalence rates within the different regions, but there is an upward trend in all the regions of the sub-Saharan Africa. This trend is believed to be related to the increasing frequencies of CAD risk factors in the subcontinent. PMID- 12764401 TI - Arm position and blood pressure: a risk factor for hypertension? AB - The objective of this study was to re-evaluate the effect of arm position on blood pressure (BP) measurement with auscultatory and oscillometric methods including ambulatory blood pressure monitoring (ABPM). The setting was the hospital outpatient department and the subjects chosen were normotensive and hypertensive. The effect of lowering the arm from heart level on indirect systolic BP (SBP) and diastolic BP (DBP) measurement as well as the importance of supporting the horizontal arm were measured. In the sitting position, lowering the supported horizontal arm to the dependent position increased BP measured by a mercury device from 103+/-10/60+/-7 to 111+/-14/67+/-10 mmHg in normotensive subjects, a mean increase of 8/7 mmHg (P<0.01). In hypertensive subjects, a similar manoeuvre increased BP from 143+/-21/78+/-17 to 166+/-29/88+/-20 mmHg, an increase of 23/10 mmHg (P<0.01). Combined results from normotensive and hypertensive subjects demonstrate a direct and proportional association between BP (SBP and DBP) and the increase produced by arm dependency. Similar changes and associations were noted with oscillometric devices in the clinic situation. However, supporting the horizontal arm did not alter BP. Of particular interest, analysis of 13 hypertensive subjects who underwent ABPM on two occasions, once with the arm in the 'usual' position and once with the arm held horizontally for BP measurement during waking hours, demonstrated changes comparable to the other devices. The mean 12-hour BP was 154+/-19/82+/-10 mmHg during the former period and significantly decreased to 141+/-18/74+/-9 mmHg during the latter period (P<0.01). Regression analysis of the change in SBP and DBP with arm position change again demonstrated a close correlation (r(2)=0.8113 and 0.7273; P<0.001) with the artefact being larger with higher systolic and diastolic pressures. In conclusion, arm movements lead to significant artefacts in BP measurement, which are greater, the higher the systolic or diastolic pressure. These systematic errors occur when using both auscultatory and oscillometric (clinic and ABPM) devices and might lead to an erroneous diagnosis of hypertension and unnecessary medication, particularly in individuals with high normal BP levels. Since clinical interpretations of heart level vary, the horizontal arm position should be the unambiguous standard for all sitting and standing BP auscultatory and oscillometric measurements. PMID- 12764402 TI - Using a problem detection study (PDS) to identify and compare health care provider and consumer views of antihypertensive therapy. AB - The objectives of this study were to ascertain consumer knowledge and behaviour about hypertension and treatment and to compare these with health care providers' perceptions (of 'most' consumers). The design for the study was a problem detection study (PDS): focus groups and then survey. Focus groups and survey participants were convenience samples of consumers, doctors, nurses and pharmacists. The main outcome measures were agreement on a 5-point Likert scale with statements about consumers' knowledge and behaviour about high blood pressure and medication. The survey identified areas of consensus and disagreement between consumers and health providers. While general knowledge and concordance with antihypertensive therapy among consumers was good, consequences such as eye and kidney disease, interactions with herbal medicines, and how to deal with missing a dose were less well known. Side effects were a problem for over one-quarter of participants, and cost was a problem in continuing therapy. Half the consumers had not received sufficient written information. Providers overall disagreed that most consumers have an adequate understanding of the condition. They agreed that most consumers adhere to therapy and can manage medicines; and about their own profession's role in information provision and condition management. Consumers confirmed positive provider behaviour, suggesting opportunities for greater communication between providers about actions taken with their consumers. In conclusion, the PDS methodology was useful in identifying consumer opinions. Differences between consumer and provider responses were marked, with consumers generally rating their knowledge and behaviour above providers' ratings of 'most' consumers. There are clear gaps to be targeted to improve the outcomes of hypertension therapy. PMID- 12764403 TI - Analysis of the regional pulse wave velocity by Doppler: methodology and reproducibility. AB - Increased arterial stiffness is observed in a number of cases. The analysis of the regional functional arterial properties is of interest to determine the role of a given risk factor on the vascular wall and in some diseases such as atherosclerosis. We analysed the pulse wave velocity (PWV) measured by the Doppler method with 2D guidance and its reproducibility in different arterial segments in 15 men with coronary artery disease. Regional Doppler PWV was defined as the distance between the extremities of a given segment divided by the transit time calculated by Doppler. Intra- and interobserver reproducibilities of the Doppler measurements were studied in all of the subjects. The variation coefficients were low, maximum at the level of ascending aorta and minimal at the level of iliac segment. This good reproducibility was validated by the Bland Altman method. Moreover, using this Doppler technique, we found a progressive increase in PWV from the ascending aorta to the iliac segment. These data demonstrate that noninvasive Doppler analysis is a feasible and reproducible method to determine regional PWV. PMID- 12764404 TI - How well are hypertension and albuminuria treated in type II diabetic patients? AB - The purpose of this study was to assess the management of both hypertension and micro/macroalbuminuria in a cohort of type II diabetic patients. In the first 6 months of the year 2000, 5815 diabetic patients were identified through prescriptions for antidiabetic drugs in our sanitary district (191 568 inhabitants). In all, 65% (3810) of these type II diabetic patients were also given prescriptions for antihypertensive drugs. A total of 400 diabetic patients were randomly selected and 171 entered the study (gender: 94/77 M/F; age: 66.6+/ 8 years; diabetes duration: 12+/-9 years): 100 patients (group DT) were treated with antihypertensive drugs and 71 (group DU) were untreated. Blood pressure, urine albumin-to-creatinine ratio (ACR), and glycated haemoglobin were measured in the two groups. A total of 80% (57/71) of DU patients were hypertensive (BP>/=130/85 mmHg). Specifically, 24.4% had diastolic hypertension (BP>/=85 mmHg) and 79% systolic hypertension (BP>/=130 mmHg). Only 63% (100/157) of the hypertensive patients were treated with antihypertensive drugs (two drugs/patient on average, range 1-5). In addition, only 13% of the DT patients were adequately controlled (BP<130/85 mmHg), while the others had above target blood pressure levels (14%: 130-139/85-89 mmHg; 40%: 140-159/90-95 mmHg, and 33%>/=160/95 mmHg). Angiotensin converting enzyme inhibitors (ACE-I) and angiotensin receptor blockers (ARB) were included in the antihypertensive medical regimen in 70% of the DT patients (ACE-I: 62%; ARB: 8%; diuretics: 39%; dihydropyridine calcium antagonists: 38%; alpha-blockers: 20%, beta-blockers: 17%; clonidin: 8%; nondihydropyridine calcium antagonists: 5%). Only 33% of type II diabetic patients underwent a screening for microalbuminuria as assessed on clinical records. The same percentage of micro- and macroalbuminuric patients (13.5%) was observed in the DT group, whereas 25% micro vs 3% macro were found in the DU group. In all, 73% of microalbuminuric patients were not on ACE-I/ARB. Hypertensive type II diabetic patients were often left untreated and only a minority of those treated were optimally controlled. The importance of an elevated systolic pressure is underestimated and the number of antihypertensive drugs prescribed insufficient. Screening and treatment of albuminuria are inadequate. PMID- 12764405 TI - Diarrhoea, vomiting and ACE inhibitors:--an important cause of acute renal failure. AB - The occurrence of severe acute renal failure in 3 patients who developed diarrhoea while taking angiotensin converting enzyme (ACE) inhibitors led us to undertake a retrospective cohort survey to determine the frequency with which diarrhoea and vomiting are associated with acute renal failure in patients taking this class of drug. Serum creatinine was measured as part of the diagnostic workup of 2398 consecutive admissions to an acute medical receiving unit in a district general hospital. Outcome measures were the presence of diarrhoea and/or vomiting, and whether taking an ACE inhibitor, NSAID or diuretic at the time of admission, also previous, initial and follow up serum creatinine concentrations. Peak serum creatinine in the 3 cases was 1159, 989 and 765 micromol/l. None of the 3 required dialysis and all recovered renal function completely after receiving large volumes of intravenous fluid. In the cohort study, 89 of 2398(3.7%) admissions had serum creatinine >/=200 micromol/l. Nine were regular dialysis patients. Of the remaining patients, 30 (37.5%) were taking an ACE inhibitor. Six of 30 (20%) gave a history of diarrhoea and/or vomiting. Median creatinine concentration in this group was 135 (range 111-209) micromol/l before admission, 292 (216-724) micromol/l when first seen in hospital, and 134 (94-219) micromol/l following the withdrawal of drug therapy and fluid replacement. In conclusion, volume depletion causing acute renal failure in patients taking ACE inhibitors is not uncommon. Such patients and their general practitioners should be aware that reversible renal impairment may occur during intercurrent illnesses, particularly if characterised by diarrhoea and/or vomiting. PMID- 12764407 TI - Perindopril and candesartan comparative efficacy and safety in type II diabetic hypertensive patients. PMID- 12764406 TI - Antihypertensive efficacy and safety of olmesartan medoxomil compared with amlodipine for mild-to-moderate hypertension. AB - The antihypertensive efficacy of the angiotensin II receptor blocker olmesartan medoxomil has been shown to compare favourably with that of other antihypertensive agents. This randomized, double-blind study compared the antihypertensive efficacy of the starting dose of olmesartan medoxomil with that of the calcium channel blocker amlodipine besylate (amlodipine) in subjects with mild-to-moderate hypertension. Following a 4-week, single-blind, placebo run-in period, 440 subjects aged >/=18 years were randomized to the starting dose of olmesartan medoxomil (20 mg/day), amlodipine (5 mg/day), or placebo for 8 weeks. Subjects were evaluated by 24-h ambulatory blood pressure monitoring (ABPM) and by seated cuff blood pressure (BP) measurements at trough. The primary end point was the change from baseline in mean 24-h diastolic blood pressure (DBP) by ABPM at Week 8. Secondary end points included change from baseline in mean 24-h ambulatory systolic blood pressure (SBP) at 8 weeks, change from baseline in mean seated trough cuff DBP and SBP measurements, and response and control rates for DBP <90 and <85 mmHg. Control rates for SBP <140 and <130 mmHg were also calculated. Olmesartan medoxomil and amlodipine produced significantly greater reductions in ambulatory and seated DBP and SBP compared with placebo. Mean reductions in ambulatory and seated BP were similar between the two active agents; however, in the olmesartan medoxomil group, significantly more patients achieved the SBP goal of <130 mmHg and the DBP goal of <85 mmHg. Both drugs were well tolerated at the recommended starting dose. Although amlodipine was associated with a higher incidence of oedema, this did not reach statistical significance. Olmesartan medoxomil is an effective antihypertensive agent, with BP-lowering efficacy at the starting dose similar to that of amlodipine, and is associated with more patients achieving the rigorous BP goals of SBP <130 mmHg and DBP <85 mmHg. PMID- 12764408 TI - Adherence to an intensive antihypertensive follow-up programme. PMID- 12764409 TI - Structural and functional cardiovascular abnormalities in never-treated hypertensives according to ACE-gene polymorphism. PMID- 12764410 TI - Genome evolution: let's stick together. PMID- 12764417 TI - Estimating the number, frequency, and dominance of S-alleles in a natural population of Arabidopsis lyrata(Brassicaceae) with sporophytic control of self incompatibility. AB - In homomorphic plant self-incompatibility (SI) systems, large numbers of alleles may be maintained at a single Mendelian locus. Most estimators of the number of alleles present in natural populations are designed for gametophytic self incompatibility systems (GSI) in which the recognition phenotype of the pollen is determined by its own haploid genotype. In sporophytic systems (SSI), the recognition phenotype of the pollen is determined by the diploid genotype of its parent, and dominance differs among alleles. We describe research aimed at estimates of S-allele numbers in a natural population of Arabidopsis lyrata (Brassicaceae), whose SSI system has recently been described. Using a combination of pollination studies and PCR-based identification of alleles at a locus equivalent to the Brassica SRK gene, we identified and sequenced 11 putative alleles in a sample of 20 individuals from different maternal seed sets. The pollination results indicate that we have not amplified all alleles that must be present. Extensive partial incompatibility, nonreciprocal compatibility differences, and evidence of weakened expression of SI in some genotypes, prevent us from determining the exact number of missing alleles based only on cross pollination data. Although we show that none of the theoretical models currently proposed is completely appropriate for estimating the number of alleles in this system, we estimate that there are between 13 and 16 different S-alleles in our sample, probably between 16 and 25 alleles in the population, and discuss the relative frequency of alleles in relation to dominance. PMID- 12764418 TI - Genetic variability of the marine mussel Mytilus galloprovincialis assessed using two-dimensional electrophoresis. AB - Two-dimensional electrophoresis (2-DE) has been used to measure the degree of genetic variability of the marine mussel Mytilus galloprovincialis. Genetic polymorphisms were detected in 33 of a total of 86 polypeptides scored among the most abundant proteins from foot samples in 38 individuals. Estimates of average heterozygosity were 0.101+/-0.018 and 0.114+/-0.021 in a natural and a cultured population, respectively, from the NW of the Iberian Peninsula. These are the highest estimates of average heterozygosity reported by 2-DE in an animal species to date. We consider that these data throw open the question of the level of genetic variability detectable by two-dimensional electrophoresis. Multilocus genotype data were used to infer haplotypic frequencies by means of the EM algorithm in order to detect linkage disequilibrium between loci coding abundant proteins. Significant associations were found in 22.7% of the 406 two-locus pairs analysed. Also, clusters of loci in which all pairwise combinations exhibit statistically significant associations were detected and physical linkage between some of these loci is postulated from the linkage disequilibrium data. PMID- 12764419 TI - The genetic structure of a columnar cactus with a disjunct distribution: Stenocereus gummosus in the Sonoran desert. AB - Stenocereus gummosus is a columnar cactus endemic to the Sonoran desert that exhibits a disjunct distribution: it is widely distributed in Baja California and restricted to a small coastal area in mainland Sonora. In this paper, we examine the genetic structure and the mating system of this species in order to explore the origin of the disjunction and describe aspects of the pollination biology. Flowers are nocturnal, pollinated mainly by sphingids and self-incompatible. Polymorphism for allozymes (11 loci) was relatively high (P=75%) but moderate levels of heterozygosity were detected (H(o)=0.103 and H(e)=0.261). Sonoran populations exhibited higher levels of genetic variation than peninsular populations. H(e) declined with latitude when just peninsular and two island populations are included. Substantial levels of inbreeding within populations (f=0.60), moderate differentiation among populations (theta=0.10), and no evidence of isolation by distance were detected. The neighbor-joining phenogram showed Sonoran and island populations nested within peninsular populations. Mainland populations showed greater genetic similarity to island populations, supporting a dispersal hypothesis for the origin of the disjunction. Future studies using DNA markers are suggested in order to better understand the forces that have shaped the genetic structure of this species. PMID- 12764420 TI - Population genetic structure in a human-disturbed environment: a case study in the land snail Helix aspersa (Gastropoda: Pulmonata). AB - Local patterns of genetic variation were analysed in the land snail Helix aspersa for 32 populations sampled within a patchy agricultural landscape: the polders of the Bay of Mont-Saint-Michel (France). This investigation examined the allele frequencies at four enzymatic markers and five microsatellite loci through the genotyping of 580 individuals. A strongly significant population genetic substructuring (mean F(ST)=0.088, P<0.001) was found at the scale of the whole polders area (3050 ha) and both categories of markers displayed a similar magnitude of spatial genetic differentiation. We did not find any obvious effects of habitat fragmentation on the distribution of genetic variability. Despite the reality of habitat patchiness and environmental instability (related to farming practices), an isolation by distance process was clearly depicted, although selective pressures cannot be ruled out for one enzymatic locus. Overall, genetic drift, along with occasional long-distance episodes of gene flow, was presumably the most likely evolutionary force that shaped the observed pattern of genetic variation. PMID- 12764421 TI - Mapping viability loci using molecular markers. AB - In genetic mapping experiments, some molecular markers often show distorted segregation ratios. We hypothesize that these markers are linked to some viability loci that cause the observed segregation ratios to deviate from Mendelian expectations. Although statistical methods for mapping viability loci have been developed for line-crossing experiments, methods for viability mapping in outbred populations have not been developed yet. In this study, we develop a method for mapping viability loci in outbred populations using a full-sib family as an example. We develop a maximum likelihood (ML) method that uses the observed marker genotypes as data and the proportions of the genotypes of the viability locus as parameters. The ML solutions are obtained via the expectation maximization algorithm. Application and efficiencies of the method are demonstrated and tested using a set of simulated data. We conclude that mapping viability loci can be accomplished using similar statistical techniques used in quantitative trait locus mapping for quantitative traits. PMID- 12764423 TI - Almotriptan in the treatment of migraine. AB - Almotriptan is a new selective serotonin 5-HT(1B/1D) receptor agonist which is chemically related to sumatriptan and is used in the acute treatment of migraine. Almotriptan, like the rest of the triptans, acts by inducing vasoconstriction of the meningeal arteries. The new drug has good oral bioavailability, and in clinical studies has been shown to be at least as effective than sumatriptan 100 mg in alleviating migraine headache and associated symptoms (nausea, vomiting, phonophobia and photophobia) when administered as a single oral dose of 12.5 mg, this being the recommended dose. However, almotriptan has a very good tolerability profile, which has been shown to be superior to that of sumatriptan in two comparative trials. Therefore, almotriptan offers clear advantages over sumatriptan in the acute treatment of migraine attacks. (c) 2001 Prous Science. All rights reserved. PMID- 12764422 TI - Chemical defence in a sawfly: genetic components of variation in relevant life history traits. AB - Larvae of several tenthredinid sawfly species readily release droplets of haemolymph through their integument when attacked by predators. This defence mechanism via 'bleeding' is characterised by a low integument resistance and a high haemolymph deterrence. Both traits are variable, and negatively correlated among species. We sought to determine if such differences in the propensity to bleed also occur intraspecifically by studying the heritability of traits potentially associated with the bleeding phenomenon in the turnip sawfly Athalia rosae ruficornis Jakovlev (Hymenoptera: Tenthredinidae, Allantinae). For three European populations, heritabilities were estimated in the laboratory in a parent offspring and a full-sib design for haemolymph deterrence (measured as concentration of sequestered glucosinolate), integument resistance, body mass of eonymph and adult, and developmental time. Within A. rosae, no significant negative phenotypic correlation was found between the two traits directly related to the defence mechanism: integument resistance and haemolymph deterrence. However, the significant heritabilities found for these traits in the full-sib analysis (0.39 and 0.35, respectively, for males in the Swiss population) show that the variation has a genetic component. While full-sib analysis revealed highly significant heritabilities for most traits in all the three populations, parent-offspring regression revealed little or no evidence of heritable variation. Effects of common environment for siblings and variation in the host plant quality between insect generations are likely to be the main factors explaining these differences. A consequence of such host-plant variation in the wild might be that genetic variation of such chemical defensive traits is largely invisible to natural selection. PMID- 12764424 TI - Antimicrobial selection in the penicillin-allergic patient. AB - Patients frequently state that they have a penicillin allergy that often presents a therapeutic problem in treating a variety of infectious disorders. Penicillin and beta-lactam allergic reactions should be determined by a careful history. Many patients who say they have a penicillin allergy, in fact do not. If it is determined that the patient has a penicillin allergy, then the clinician should determine whether it is of an anaphylactic or nonanaphylactic variety. Most reactions to beta-lactams are of the nonanaphylactic variety and are usually manifested clinically as a mild maculopapular rash or drug fever. Uncommonly, penicillin allergies are clinically manifested as anaphylactic reactions, e.g., bronchospasm, laryngospasm, hypotension or hives. Patients' hypersensitivity reactions tend to be stereotyped on rechallenge, which make the reactions predictable. Patients who have a questionable penicillin allergy, or have had only fever or rash, may be safely given beta-lactam antibiotics without fear of anaphylaxis. Patients with a documented history of anaphylactic reactions should receive non-beta-lactam antibiotics. Although monobactams and carbapenems are structurally related to beta-lactams, they are unrelated in terms of allergic potential. There is no cross-reactivity between mono-bactams or carbapenems with beta-lactams, and these drugs may be used safely in patients with anaphylactic reactions to beta-lactams. Because so many antibiotics are available that are allergically unrelated to beta-lactams, beta-lactam desensitization procedures are rarely necessary. (c) 2001 Prous Science. All rights reserved. PMID- 12764425 TI - Combination treatment with sirolimus and ciclosporin in clinical renal transplantation: A comprehensive review. AB - Sirolimus is a potent new immunosuppressive agent that has been shown to reduce the incidence of acute rejection episodes among renal transplant recipients as well as provide a unique approach to optimize treatment outcomes in difficult transplant situations. Owing to its properties as a critical-dose drug, therapeutic concentration monitoring of sirolimus readily compensates for intra- and interpatient variability and drug interactions with a variety of other agents such as ciclosporin. This review summarizes the results that demonstrate the efficacy of sirolimus in combination treatment with ciclosporin in human renal transplantation, as well as its potential in alternative therapeutic modalities in a broad range of transplant recipients. The clinical trials for SDZ-RAD, a macrocyclic lactone immunosuppressant structurally similar to sirolimus, also are reviewed herein. SDZ-RAD was developed in an attempt to improve the pharmacokinetic characteristics of sirolimus, particularly to increase the extent and reproducibility of its oral bioavailability and to reduce the extensive tissue distribution by virtue of its greater polarity. (c) 2001 Prous Science. All rights reserved. PMID- 12764426 TI - Gemifloxacin. AB - Gemifloxacin is a novel antibiotic and the first fluoroquinolone with a pyrrolidine derivative at the C-7 position. Because of the added pyrrolidine substitute, gemifloxacin has an enhanced spectrum of activity against Gram positive bacteria such as Streptococcus pneumoniae and Staphylococcus aureus, in addition to its activity against Gram-negative bacteria. Like other fluoroquinolones, gemifloxacin's mechanism of action focuses on inhibiting DNA gyrase and topoisomerase, thus preventing cellular replication. In addition, in vitro and in vivo data have shown that the compound exhibits excellent activity against Enterobacteriaceae and other respiratory pathogens. Furthermore, it has been demonstrated that gemifloxacin has potential activity in vitro against anaerobic bacteria. With a broad spectrum of activity, convenient once-daily administration, good bio-availability and tolerability, gemifloxacin will be an important addition to our armamentarium against a wide range of infections, from urinary tract infections to community-acquired pneumonia. (c) 2001 Prous Science. All rights reserved. PMID- 12764427 TI - Nateglinide: A structurally novel, short-acting, hypoglycemic agent. AB - Nateglinide is a short-acting, pancreatic, beta-cell-selective, K(ATP) potassium channel blocker that improves overall glycemic control in type 2 diabetes. Although nateglinide's mechanism of action is related to that of sulphonyl-ureas and repaglinide, important differences do exist. Nateglinide binds rapidly to the sulfonylurea SUR1 receptor with a relatively low affinity, and it dissociates from it extremely rapidly in a manner of seconds. This rapid association and dissociation gives nateglinide a unique "fast on-fast off" effect. Thus, nateglinide has a rapid onset and short duration of action stimulating insulin secretion in vivo and providing good control of postprandial hyperglycemia when taken immediately prior to meals. The rapid action of nateglinide on the beta cells stimulates and restores the normal physiological first and early phase of insulin secretion, consequently reducing postprandial hyperglycemia. This hypoglycemic effect of nateglinide leads to improved glycemic control, while the short duration avoids delayed hyperinsulinemia and hypoglycemia after meals. Nateglinide is not a sulfonylurea, but it shares the mechanism of action of commonly used oral hypoglycemic agents such as glibenclamide and glipizide. Like the recently introduced, short-acting agent, repaglinide, it does not incorporate a sulfonylurea moiety. However, nateglinide's effects on insulin secretion and glycemic control differ significantly from the sulfonylureas and repaglinide in that it preferentially stimulates acute phase insulin, better controls postprandial glucose excursions and spikes, and causes less hyperinsulinemia and hypoglycemia. Compounds with such a profile should not only achieve improved overall glucose control, but also reduce the risk of vascular complications which is the most important feature of nateglinide. Clinical studies with nateglinide have confirmed that it acts rapidly and both restores insulin release and attenuates the postprandial glucose spike. Nateglinide is both effective and well tolerated in the treatment of type 2 diabetes. The reported overall profile of adverse effects appears to be superior to that of other K(ATP) potassium channel blockers, the glucose modulator metformin and PPARgamma agonists such as troglitazone. Clinical comparisons of these agents have shown nateglinide to be more effective in attenuating postprandial glucose than any other oral hypoglycemic agent, and that treatment with both nateglinide and metformin provides additive effects that afford improved control of plasma glucose levels. The administration regimen for nateglinide, immediately prior to meals, also facilitates patient compliance. (c) 2001 Prous Science. All rights reserved. PMID- 12764428 TI - Pharmacologic agents in complementary medicine in prostatic disease. AB - The use of alternative medicine has increased considerably in the last 10 years. Some of the reasons for this trend include limited efficacy of existing treatments, perceived reduction in side effects with alternative treatments, patient's desire to maintain control over their treatment and a desire for a more "natural" treatment. The very high frequency of urologic diseases such as benign prostatic hyperplasia and prostate cancer makes these diseases natural targets of alternative medicine. Although a great number of therapeutic agents have been recommended for urologic diseases, this review examines the most promising for their use in benign prostatic hyperplasia and prostate cancer. (c) 2001 Prous Science. All rights reserved. PMID- 12764429 TI - Cell fractionation of parasitic protozoa: a review. AB - Cell fractionation, a methodological strategy for obtaining purified organelle preparations, has been applied successfully to parasitic protozoa by a number of investigators. Here we present and discuss the work of several groups that have obtained highly purified subcellular fractions from trypanosomatids, Apicomplexa and trichomonads, and whose work have added substantially to our knowledge of the cell biology of these parasites. PMID- 12764430 TI - Temporal and spatial distribution of Leishmania mexicana infections in a population of Neotoma micropus. AB - A 19-month mark-release-recapture study of Neotoma micropus with sequential screening for Leishmania mexicana was conducted in Bexar County, Texas, USA. The overall prevalence rate was 14.7% and the seasonal prevalence rates ranged from 3.8 to 26.7%. Nine incident cases were detected, giving an incidence rate of 15.5/100 rats/year. Follow-up of 101 individuals captured two or more times ranged from 14 to 462 days. Persistence of L. mexicana infections averaged 190 days and ranged from 104 to 379 days. Data on dispersal, density, dispersion, and weight are presented, and the role of N. micropus as a reservoir host for L. mexicana is discussed. PMID- 12764431 TI - Human papillomavirus prevalence among women with cervical intraepithelial neoplasia III and invasive cervical cancer from Goiania, Brazil. AB - This study estimated the prevalence and distribution of human papillomavirus (HPV) types among women with cervical intraepithelial neoplasia (CIN) grade III and invasive cervical cancer from Goi s (Brazil Central Region). Seventy-four cases were analyzed and consisted of 18 CIN III, 48 squamous cell carcinomas, 4 adenocarcinomas, 1 adenosquamous carcinoma and 3 undifferentiated carcinomas. HPV DNA sequences were examined in formalin-fixed and paraffin-embedded tissues using primers from L1 region GP5+/GP6+. Polymerase chain reaction products were typed with dot blot hybridization using probes for HPV 16, 18, 31, 33, 45, 54, 6/11, 42/43/44, 51/52, 56/58. The prevalence of HPV was estimated to be 76% (56/74). HPV 16 was the most frequently found type, followed by HPV 33, 18 and 31. The prevalence of untyped HPV was 6%; 79% percent of the squamous cell carcinoma cases and 61% percent of the CIN III were positive for HPV and the prevalence rate of HPV types was the same for the total number of cases. According to other studies, HPV type 16 is the most prevalent virus in all Brazilian regions, but there is variation regarding to other types. Type 18 is the second most prevalent HPV in North, Southeast and South Brazil regions and types 31 and 33 are the second most prevalent HPV in Northeast and Central Brazil, respectively. PMID- 12764432 TI - Phlebotominae sand flies in Paraguay: abundance distribution in the Southeastern region. AB - From September 1993 to August 2001, 7,190 phlebotomine were collected with CDC light trap in an endemic area for human leishmaniasis, in the departments of Misiones and Itap a, Paraguay. Eleven species were identified: Lutzomyia neivai (93.7%), L. whitmani (4.1%), and L. fischeri, L. shannoni, L. migonei, L. misionensis, L. cortelezzii, L. pessoai, L. alphabetica, Brumptomyia avellari and B. guimaraesi (less than 1%). The last three species are new records for the country. The biodiversity and phlebotomine abundance were associated with the proximity to primary forest or gallery forest, but L. neivai was also found in peridomestic periurban environment. L. neivai was found throughout the year, and showed a period of higher activity from September to April (spring to fall) with a unimodal or bimodal pattern in relation to the annual rainy peaks during the summer. Background literature about phlebotomine from Paraguay has been reviewed. PMID- 12764433 TI - Dispersal of Aedes aegypti and Aedes albopictus (Diptera: Culicidae) in an urban endemic dengue area in the State of Rio de Janeiro, Brazil. AB - Experimental releases of female Aedes (Stegomyia) aegypti and Aedes (Stegomyia) albopictus were performed in August and September 1999, in an urban area of Nova Igua u, State of Rio de Janeiro, Brazil, to estimate their flight range in a circular area of 1,600 m where 1,472 ovitraps were set. Releases of 3,055 Ae. aegypti and 2,225 Ae. albopictus females, fed with rubidium (Rb)-marked blood and surgically prevented from subsequent blood-feeding, were separated by 11 days. Rb was detected in ovitrap-collected eggs by atomic emission spectrophotometry. Rb marked eggs of both species were detected up to 800 m from the release point. Eggs of Ae. albopictus were more numerous and more heterogeneously distributed in the area than those of Ae. aegypti. Eggs positively marked for Rb were found at all borders of the study area, suggesting that egg laying also occurred beyond these limits. Results from this study suggest that females can fly at least 800 m in 6 days and, if infected, potentially spread virus rapidly. PMID- 12764434 TI - Mortality in immatures of the floodwater mosquito Ochlerotatus albifasciatus (Diptera: Culicidae) and effects of parasitism by Strelkovimermis spiculatus (Nematoda: Mermithidae) in Buenos Aires Province, Argentina. AB - Life tables were constructed for six cohorts of immature stages of the floodwater mosquito Ochlerotatus albifasciatus (Macquart) in a park in Buenos Aires, highlighting the mortality attributable to the parasitic nematode, Strelkovimermis spiculatus Poinar & Camino. Two cohorts were selected to compare parasite incidence in all mosquito stages when low and high parasitism occurred. Development time of Oc. albifasciatus from first instar to adult was 7.7-10 days in the spring, 6 days in the summer, and 10.9-21.9 days in the fall. Survival was estimated as 0-1.4% in the spring, 2% in the summer and 0.2-4.4% in the fall. The highest "K" value (Killing power) occurred during a fall cohort when prevalence of the parasite was 86.9%, and the lowest in a spring cohort. Parasitism occurred during all seasons, but S. spiculatus persisted to adult only in the summer and fall, when adult mosquitoes developed from parasitized third and fourth instars larvae. The abundance of S. spiculatus differed between old and young larvae only when parasite prevalence was the highest. Although pupae and adults of Oc. albifasciatus were parasitized, no pupal mortality attributable to parasitism was recorded. The proportion of parasitized adults ranged from 14.2% and 5.7% in the two cohorts compared. Pupal wet weight and adult wing lengths did not differ between parasitized and unparasitized individuals. PMID- 12764435 TI - Anti-human immunodeficiency virus-1 antibody titers in injection drug users compared to sexually infected individuals. AB - Sera from infected injection drug users (IDU) have shown to have antibodies against synthetic human immunodeficiency virus-1 (HIV-1) envelope peptides more frequently. In this study, reactivity of 48 IDU plasma were compared to 60 plasmas obtained from sexually infected individuals (S). The overall reactivity of plasma from IDU compared to S was higher, and the reactivity titers were much higher for IDU plasma than S. IDU plasma also showed a broader antibody response. The higher reactivity titers were observed mainly for the gp41 immunodominant epitope and V3 peptides corresponding to the consensus sequences of HIV-1 subtypes/variants prevalent in Brazil (B, F, C) indicating the specificity in the higher immune response of IDU. PMID- 12764436 TI - Quantifying the potential pathogens transmission of the blowflies (Diptera: Calliphoridae). AB - To quantify the potential capability of transporting and passing infective pathogens of some blowflies (Diptera: Calliphoridae), Mihalyi's danger-index was calculated for seven species. The original equation was modified to include synanthropic information to discriminate between asynanthropic, hemisynanthropic, and eusynanthropic status. Three groups were recognized, of which Phaenicia cluvia and Musca domestica proved the flies with lowest index value (D = 2.93 and 3.00 respectively); Cochliomyia macellaria, Chrysomya albiceps and Sarconesia chlorogaster presented a significantly higher index value (p<0.10; D = 4.28, 4.44 and 5.66 respectively) and C. megacephala, C. vicina and P. sericata appear to represent the heaviest potential sanitary risk with the highest index value (p<0.10; D = 15.54, 16.88 and 12.49 respectively). PMID- 12764437 TI - Reproductive aspects of the mosquito Culex quinquefasciatus (Diptera:Culicidae) infected with Wuchereria bancrofti (Spirurida: Onchocercidae). AB - This study reports on the relationship between Wuchereria bancrofti infection and female body size, intake of blood and fecundity in the mosquito Culex quinquefasciatus, vector of this filarial parasite in Recife (Brazil). Adults from field collected larvae were infected via a membrane feeding procedure, using blood with parasitaemia ranging from 724-6,000 mf/ml. A positive correlation was observed between mosquito size (measured by wing length) and egg production in uninfected females. However, this relationship did not exist in W. bancrofti infected mosquitoes. This change is unlikely to be the result of changes in blood ingestion as no significant difference was found when infected and uninfected females were compared. Variation in egg production observed between trials could not be associated with parasite density in the blood. These results suggest infection with W. bancrofti may disrupt the relationship between mosquito size and egg production during the first gonotrophic cycle of C. quinquefasciatus such that fecundity is sometimes reduced. However, this overall affect is variable and many groups of mosquitoes do not respond in this way. PMID- 12764438 TI - Oviposition response of the mosquito, Culex quinquefasciatus to the secondary metabolite(s) of the fungus, Trichoderma viride. AB - Secondary metabolites produced by Trichoderma viride, a deuteromycetes fungus, under submerged culture condition were formulated and evaluated for oviposition attractancy against gravid females of Culex quinquefasciatus mosquito. At a concentration of 10 g ml-1 the formulation showed remarkable attractancy with an oviposition active index (OAI) of +0.52. When the oviposition attractancy of the formulation was compared with a known oviposition attractant, p-cresol, both at 10 g ml-1, the former was found to be more attractive to result in 70% egg laying than the later with 30% egg laying. Thin layer chromatography fractions of the secondary metabolites showed that a fraction with Rf value of 0.88 was highly active as oviposition attractant with an OAI of +0.65. Further work on identification of the active principle(s) of the microbial formulation might lead to an oviposition attractant useful in mosquito vector management. PMID- 12764439 TI - Influence of the blood meal source on the biology of Meccus picturatus Usinger 1939 (Hemiptera: Reduviidae: Triatominae) under laboratory conditions. AB - Aspects related to hatching, time-lapse between presenting the blood meal and beginning of feeding, feeding time, postfeed defecation delay,life time, mortality and fecundity for each stage of Meccus picturatus, life-cycle were evaluated and compared in two cohorts of M. picturatus fed on hens or rabbits. The hatching rate observed for each of the two studied groups of eggs was 78.1% (n = 2298) on the group fed on hens and 82.1% (n = 2704) on that fed on rabbits, and the average time of hatching was 20 days. Mean time-lapse for beginning feeding was under 3 min in nymphal stages and postfeed defecation delay was under 10 min in all stages, in both cohorts. Mean feeding time was significantly (P < 0.05) shorter in triatomines fed on hens than on rabbits. A similar number of nymphs of each cohort, 69 fed on hens (34.5%) and 68 fed on rabbits (34%), completed the cycle. No significantly (P > 0.05) differences were recorded among the average times from NI to adult in the cohort fed on hens (196.8 15.8 days) and the average time in the cohort fed on rabbits (189.5 22.9). The average span in days for each stage fed on hens was not significantly different to the average span for each stage fed on rabbits. The number of blood meals at each nymphal stage varied from 1 to 6 in both cohorts. The mortality rates were higher on fifth nymphal stage, in both cohorts. No significant (P > 0.05) differences were recorded on mortality rates on most nymphal stages of both cohorts. The average number of eggs laid per female from the cohort fed on hens in a 9-month period was 791.1, whereas the average number of eggs in the cohort fed on rabbits was 928.3. PMID- 12764440 TI - The type specimens of chewing lice (Insecta, Mallophaga) deposited in the entomological collection of Instituto Oswaldo Cruz, Rio de Janeiro, RJ, Brazil. AB - This paper presents a list of 131 Mallophaga type specimens deposited in the Werneck Collection of Instituto Oswaldo Cruz, Rio de Janeiro, RJ, Brazil. It includes 69 holotypes, 62 allotypes, 683 paratypes, 1 syntype and 2 neoparatypes, distributed among 6 families and 35 genera. The types are listed with their respective data and literature. PMID- 12764441 TI - A new species of Acantholochus (Copepoda: Bomolochidae) parasitic on Centropomus undecimalis (Osteichthyes: Centropomidae) from the coastal zone of the State of Rio de Janeiro, Brazil. AB - A new species of Acantholochus Cressey, 1984 (Copepoda: Bomolochidae) parasitic on the gills of common snook, Centropomus undecimalis, from the coastal zone of the State of Rio de Janeiro, Brazil, is described and illustrated. The new species differs from all other species of Acantholochus by the presence of only one inner seta on middle segment of the second and third endopods. PMID- 12764442 TI - Isoenzymatic variability among five Anopheles species belonging to the Nyssorhynchus and Anopheles subgenera of the Amazon region, Brazil. AB - An isoenzymatic comparative analysis of the variability and genetics differentiation among Anopheles species was done in populations of An. (Nys.) intermedius and An. (Ano.) mattogrossensis of the Anopheles subgenus, and of An. darlingi, An. albitarsis and An. triannulatus of the Nyssorhynchus subgenus, with the aim of detecting differences between both subgenera and of estimating the degree of genetic intere specific divergence. Samples from Macap , State of Amap and Janauari Lake, near Manaus, State of Amazonas, were analyzed for eight isoenzymatic loci. Analysis revealed differences in the average number of alleles per locus (1.6-2.3) and heterozygosity (0.060-0.284). However, the proportion of polymorphic loci was the same for An. (Nys.) darlingi, An. (Nys.) triannulatus and An. (Ano.) mattogrossensis (50%), but differed for An. (Nys.) albitarsis (62.5%) and An. (Ano.) intermedius (25%). Only the IDH1 (P > 0.5) locus in all species studied was in Hardy-Weinberg equilibrium. The fixation index demonstrated elevated genetic structuring among species, based on values of Fst = 0.644 and genetic distance (0.344-0.989). Genetic difference was higher between An. (Nys.) triannulatus and An. (Ano.) intermedius (0.989) and smaller between An. (Nys.) albitarsis sensu lato and An. (Nys.) darlingi (0.344). The data show interspecific genetic divergence which differs from the phylogenetic hypothesis based on morphological characters. PMID- 12764443 TI - Human mixed infections of Leishmania spp. and Leishmania-Trypanosoma cruzi in a sub Andean Bolivian area: identification by polymerase chain reaction/hybridization and isoenzyme. AB - Parasites belonging to Leishmania braziliensis, Leishmania donovani, Leishmania mexicana complexes and Trypanosoma cruzi (clones 20 and 39) were searched in blood, lesions and strains collected from 28 patients with active cutaneous leishmaniasis and one patient with visceral leishmaniasis. PCR-hybridization with specific probes of Leishmania complexes (L. braziliensis, L. donovani and L. mexicana) and T. cruzi clones was applied to the different DNA samples. Over 29 patients, 8 (27.6%) presented a mixed infection Leishmania complex species, 17 (58.6%) a mixed infection Leishmania-T. cruzi, and 4 (13.8%) a multi Leishmania T. cruzi infection. Several patients were infected by the two Bolivian major clones 20 and 39 of T. cruzi (44.8%). The L. braziliensis complex was more frequently detected in lesions than in blood and a reverse result was observed for L. mexicana complex. The polymerase chain reaction-hybridization design offers new arguments supporting the idea of an underestimated rate of visceral leishmanisis in Bolivia. Parasites were isolated by culture from the blood of two patients and lesions of 10 patients. The UPGMA (unweighted pair-group method with arithmetic averages) dendrogram computed from Jaccard's distances obtained from 11 isoenzyme loci data confirmed the presence of the three Leishmania complexes and undoubtedly identified human infections by L. (V.) braziliensis, L. (L.) chagasi and L. (L.) mexicana species. Additional evidence of parasite mixtures was visualized through mixed isoenzyme profiles, L. (V.) braziliensis-L. (L.) mexicana and Leishmania spp.-T. cruzi. The epidemiological profile in the studied area appeared more complex than currently known. This is the first report of parasitological evidence of Bolivian patients with trypanosomatidae multi infections and consequences on the diseases' control and patient treatments are discussed. PMID- 12764444 TI - Use of polymerase chain reaction and enzymatic cleavage in the identification of Helicobacter spp. in gastric mucosa of human beings from North Parana, Brazil. AB - Helicobacter pylori is the most common gastric bacteria of human beings. Animal borne helicobacter have been associated with gastritis, ulceration, and gastric mucosa-associated lymphoid-tissue lymphoma in people. We attempted to identify the species of Helicobacter spp. that infect human beings in north Paran , Brazil. Samples of gastric mucosa from 38 dyspeptic patients were analyzed by optic microscopy on silver stained slides, polimerase chain reaction (PCR), and enzymatic cleavage. Genus and species-specific primers to H. pylori, H. heilmannii, H. felis, and consensual primers to H. bizzozeronii or H. salomonis were used. The PCR products were submitted to enzymatic cleavage by VspI (Helicobacter spp. product) and HinfI (species products) enzymes. Thirty-two out of 38 patients evaluated had 3.2 to 5 m long bacteria that resembled H. pylori in Warthin-Starry stained slides and were positive to the genus Helicobacter by PCR. In 30 of these patients the bacteria were identified as H. pylori. Two samples positive by silver stain were negative to all species tested by PCR. None of the 38 samples was positive to animal-origin helicobacter species. These results show that PCR and enzymatic restriction are practical methods to identify the species of helicobacters present in gastric mucosa of human beings. People in north Paran appear to be infected mostly with H. pylori. PMID- 12764445 TI - Standardization of bovine macrophage monolayers and isolation and culture of trypanosomes. AB - We describe a method for culturing over 90% pure bovine macrophages from peripheral blood mononuclear cells separated with Nycoprep. The cells were cultured for 12 days and then stained with esterase and with anti CD14 to test for purity. The method is reproducible and ensures an adequate number of cells for immunological research. Additionally, we report the unexpected finding of Trypanosoma trypomastigotes in our macrophage cultures from bovines belonging to a geographic area from which no bovine trypanosomes had been reported before. PMID- 12764446 TI - Perspective of a new diagnostic for human trichomonosis. AB - Several diagnostic techniques have been employed for the detection of Trichomonas vaginalis. Microtubules constitute the cytoskeleton in eukaryotic cells and are sensitive to antimitotic drugs, such as Taxol (paclitaxel). We used FLUTAX a fluorescent taxoid - to analyze the microtubule distribution in living trophozoites of T. vaginalis in urine and in vaginal discharge. A high intensity of fluorescence was observed in living T. vaginalis, epithelial cells and leukocytes present in urine and vaginal discharge. Our preliminary results show the perspective of a new diagnostic technique for trichomonosis and will contribute to the understanding of the cytoskeleton of T. vaginalis. PMID- 12764447 TI - Impact of residual spraying on Rhodnius prolixus and Triatoma dimidiata in the department of Zacapa in Guatemala. AB - As a vector control program to control Chagas disease in Guatemala, residual spraying of Rhodnius prolixus and Triatoma dimidiata was performed, and its impact was measured in the department of Zacapa. In order to identify infested villages and determine the degree of infestation, a baseline entomological survey to identify municipalities infested with vectors followed by an additional vector survey in areas known to be infested was conducted. Residual spraying using pyrethroid insecticides was performed at all the villages identified as being infested with the vectors. The residual spraying was shown to be highly effective against both vectors by the decrease in infestation indices after spraying. Analysis of the cost-effectiveness of the spraying showed that the average cost of insecticides per house is high when compared with that in Southern Cone countries. PMID- 12764448 TI - In vitro antibacterial activity of a 7-O-beta-D-glucopyranosyl-nutanocoumarin from Chaptalia nutans (Asteraceae). AB - Ethanolic crude extracts from the roots of Chaptalia nutans, traditionally used in Brazilian folk medicine, were screened against Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa by using the disk diffusion test technique. S. aureus with 14 mm inhibition zone was considered susceptible. E. coli and P. aeruginosa without such a zone were considered resistant. As a result of this finding, the ethanolic crude extract was fractionated on silica gel column chromatography into five fractions. The ethyl acetate fraction was active against S. aureus and Bacillus subtilis. Further column chromatography separation of the ethyl acetate fraction afforded 30 fractions, which were assayed against S. aureus. Fractions 16 and 17 showed inhibition zones with S. aureus, indicating the presence of active compounds, and were subjected to purification by repeated preparative thin layer chromatography. The pure compound 7-O-beta-D glucopyranosyl-nutanocoumarin inhibited B. subtilis and S. aureus at concentrations of 62.5 g/ml and 125 g/ml, respectively. The antibacterial property of C. nutans appears to have justified its use for the treatment of wounds, which are contaminated through bacterial infections. PMID- 12764450 TI - Research in public health in Brazil. PMID- 12764449 TI - Preparation of sand fly (Diptera: Psychodidae: Phlebotominae) specimens for histological studies. AB - Phlebotominae sand fly specimens were prepared for histological and physiological studies. Different fixatives were tested on sectioned and whole bodied adult females in order to obtain good fixation and provide satisfactory penetration of the embedding media. All fixed specimens were infiltrated (up to seven days under 5 C) and embedded in hydroxyethyl metacrylate. Two-three m sections were stained, mounted in Canada balsam and observed by light microscopy. Best results were achieved when whole bodied insects were double fixed in Bouin's and Carnoy's fluids (4 h/2 h) and stained in Hematoxilin/Eosin or fixed in calcium formaldehyde and stained in mercury bromophenol blue. PMID- 12764451 TI - [Does periodontal disease cause cardiovascular disease? Analysis of epidemiological evidences]. AB - This article reports a critical analysis of epidemiologic studies that evaluated periodontal disease as a cause of cardiovascular disease. Thirty-five studies were identified through a manual search of the special abstracts volumes of the Journal of Dental Research, as well as an electronic search on MEDLINE, LILACS, and ISI and inspection of the articles' bibliographies. Inclusion criteria were: articles in any language published between 1989 and 2000 reporting the presence or absence of an association between periodontal and cardiovascular diseases. Available studies are scarce, and interpretations are limited by potential bias and confounding. The studies analyzed (whether separately or jointly) fail to provide convincing epidemiologic evidence for a causal association between periodontal and cardiovascular diseases. Although the possibility that oral diseases can cause cardiovascular diseases cannot be discarded, until better data are available, periodontal disease should not be incriminated as a cause of cardiovascular disease. PMID- 12764452 TI - [Non-inert industrial solid waste disposal in landfill dumps: evaluation of toxicity and implications for the environment and human health]. AB - According to Brazilian recommended technical procedures (mainly NBR 10,004), solid waste must be previously classified in order to be disposed of adequately. Non-inert industrial solid waste is being dumped in landfill areas, most of which operate inefficiently and are located near the population and important ecosystems. In order to evaluate the potential toxicity of solid waste produced by various types of industries, 21 samples were analyzed according to NBR 10,004 procedures. Of these, 18 were classified as non-inert solid waste. The main substances contributing to the classification of these samples as non-inert waste were: aluminum, iron, manganese, phenol, and surfactants. Aluminum, manganese, and phenol are the main toxicologically relevant substances for human and environmental health because they can alter the quality of groundwater situated under solid waste disposal areas. PMID- 12764453 TI - [Effectiveness of weekly iron sulfate in the Family Health Program in Caruaru, Pernambuco State, Brazil]. AB - This study evaluates the effectiveness of an experimental project for combating anemia in children ages 6-23 months using weekly doses of iron sulfate within the Family Health Program. In 1999, the Brazilian Ministry of Health began an experimental program to combat anemia in 300,000 children from 512 municipalities in the Northeast of the country. Children received a weekly dose of 50mg of elementary iron for approximately 24 weeks. Two hundred and ninety-three children ages 6-23 months from the urban and rural areas of the municipality of Caruaru in Pernambuco State were evaluated. Before the trial, mean hemoglobin was 10.1 (+/- 0.98) g/dl, and 77.5% of the children were anemic. Following treatment, mean hemoglobin rose to 11.1 (+/- 0.87) g/dl, while the prevalence of anemia fell to 40.3%. This strategy represents a promising tool to reduce prevalence and to control severe forms of anemia in children. PMID- 12764454 TI - [Macro-epidemiologic aspects of schistosomiasis mansoni: analysis of the impacts of irrigation systems on the spatial profile of the endemic in Bahia, Brazil]. AB - Studies in African countries have shown an association between irrigation projects and the spread of schistosomiasis mansoni and haematobium. The aim of this study was to investigate the potential association between irrigation levels and the occurrence and spread of Schistosoma mansoni infection in the State of Bahia. Two forms of irrigation were developed in the State of Bahia: the first is capital-intensive and mechanized, requiring little manual labor. The second is labor-intensive and characterized by limited mechanization. According to the study, the municipalities with the largest irrigated areas are not the ones with the highest S. mansoni infection rates. In most of these counties, irrigation is capital- and technology intensive. According to the findings, unlike Africa, in the State of Bahia irrigation has had little impact on the spatial profile of the schistosomiasis endemic. PMID- 12764455 TI - Validation of a tool for assessing the quality of pharmaceutical services. AB - This paper presents the validation process for a tool assessing basic pharmaceutical services through an analysis of the implementation of a Basic Pharmaceuticals Distribution Program by the Brazilian Federal government. The process began with the drafting of a theoretical model, based on a state-of-the art review and allowing the selection of various conceptual dimensions and respective criteria that best represented the construct. The second step involved weighting indicators for the construction of quality scores. Three models were tested for ranking implementation levels, and seven simulations were conducted, determining the score most closely reflecting the selected indicators in two different matrices. The objective was to select the most coherent and consistent version between implementation levels and expected outcomes, while simultaneously enhancing validity of chosen criteria. Testing of the various models and the results obtained showed that augmenting the validity of the study was possible without altering data. This endeavor is justified in understanding the scope and limitations of these measurements and of the choices involved in issues concerning their weighting and interpretation. PMID- 12764456 TI - [Non-fatal work-related accidents in adolescents]. AB - This community-based survey estimates the annual incidence of non-fatal work related injuries in adolescents and describes their characteristics. Using a random area sampling design, 2,513 households were selected from the city of Salvador, capital of Bahia State, Northeast Brazil. The study population included 361 young people 10 to 20 years of age who answered individual questionnaires on socio-demographic, occupational, and health data. Annual incidence of non-fatal work-related injuries was 6.4%. Among females, most injuries were mild and were related to household chores; some 38.5% required medical care, and 36.4% caused permanent non-disabling lesions. Among males, work-related injuries took place mainly in public byways, and the majority (60.0%) were treated in emergency-care facilities. These results show that work-related injuries need to be considered in adolescent protection policies. PMID- 12764457 TI - [Prevalence of malnutrition among children in Embu, Sao Paulo State, Brazil, 1996 1997]. AB - This paper focused on the nutritional profile of children in the municipality of Embu, Sao Paulo State, Brazil, in 1996-1997, to identify vulnerable population segments that require specific action by health services. The sample consisted of 320 children < or = 5 years of age distributed into four socioeconomic strata. The indices were expressed as z-scores: weight/age (W/A), height/age (H/A), and weight/height (W/H) to analyze the nutritional status, and the reference for normality was the NCHS curve. In all population strata and age groups, the height/age index was the most frequently affected, while the weight/height index had the fewest deficits. No statistically significant differences were observed in children's nutritional status between the four population strata or between the different age groups. Children with low birth weight showed the highest prevalence of deficits in all indices. In the municipality, the frequency of deficits were: H/A< 2z: 7.1%, W/H< 2z: 0.2%, and W/A< 2z: 2.9%. Height deficit can be used as an early warning, considering that loss observed over the course of years has future consequences. PMID- 12764458 TI - Serum DDT in malaria vector control sprayers in Mato Grosso State, Brazil. AB - DDT was used intensively in vector control programs in Mato Grosso State until 1997. The present study aimed to determine DDT concentrations in blood samples from Brazilian National Health Foundation workers in Mato Grosso. Blood samples were analyzed from 41 sprayers, 20 drivers, and 14 unexposed workers, collected in June 1999 and October 2000 in two regions of the State (Sinop and C ceres). Sprayers and drivers were occupationally exposed, and no significant differences were found in serum DDT levels between these two groups in either region. Likewise, no significant differences were found in p,p'DDE and total DDT levels between C ceres and Sinop. However, p,p'DDT levels were higher in Sinop due to the intensive use of this insecticide in the region in recent years. The two regions together showed the following results: total DDT ranging from 7.50 micro g/L to 875.5 micro g/L (median = 135.5 micro g/L) for sprayers; from 34.5 micro g/L to 562.3 micro g/L (median = 147.7 micro g/L) for drivers; and from undetected to 94.8 micro g/L (median = 22.5 micro g/L) for unexposed workers. PMID- 12764459 TI - [Increase in leprosy detection rates in Sao Luis, Maranhao, Brazil, from 1993 to 1998: is the endemic expanding?]. AB - A descriptive epidemiologic study on the detection of new leprosy cases was conducted in Sao Luis, Maranhao, Brazil, from 1993 to 1998. A database was created for the purpose, covering 2,796 reported cases. General detection rates were calculated, as well as specific rates by gender, clinical type, and age group. Linear, exponential, geometric, and log adjustment models were performed to analyze time trends in the disease. An increase in detection was observed, involving mostly female and paucibacillary cases, mainly of tuberculoid leprosy. The increase in detection was most evident in the 15 to 19 year-old population. The percentage of detection under 15 indicated the need for active case search in this group. PMID- 12764460 TI - [Residual concentration of hexachlorocyclohexane in a contaminated site in Cidade dos Meninos, Duque de Caxias, Rio de Janeiro, Brazil, after calcium oxide treatment]. AB - Hexachlorocyclohexanes (HCHs) are currently considered potentially carcinogenic to humans. In Brasil, severe soil contamination by HCHs has been identified at Cidade dos Meninos, Duque de Caxias, Rio de Janeiro, where an old HCH factory was abandoned 40 years ago, leaving 1,700 local residents at risk. Calcium oxide treatment was performed on the most heavily contaminated (focus) area. Analysis of superficial soil samples collected years after treatment still showed high residual HCH. Contamination levels as high as 6,200mg/Kg and 7,320mg/Kg were found for alpha- and beta-HCH isomers. For the gamma- and delta-HCH isomers, concentrations reached up to 140mg/Kg and 530mg/Kg, respectively. The results showed that calcium oxide treatment was not efficient to decontaminate the soil in this area. PMID- 12764461 TI - [Risk factors for hospitalization due to acute diarrhea in children under two years old: a case-control study]. AB - The aim of this study was to investigate the socioeconomic, demographic, and biological determinants of hospitalization due to acute diarrhea (AD) in children under two years old. The hospital-based, case-control study was conducted from May to October 1997 in a pediatric hospital in Recife, Pernambuco, Brazil. Cases (n = 185) were children hospitalized due to AD, and controls (n = 185) were children with various diseases (except AD) presenting in the previous 15 days in the same area and time period. Data analysis used the Epi Info and Stata software. Odds ratios and 95% confidence intervals were calculated by multiple logistic regression to control confounding factors, considering a hierarchical risk factor model. Investigation using the proposed model showed an association between hospitalization due to AD and packed earth or plank floors in the house, greater multiparity, age of child under six months, and severity of bouts of diarrhea, which depended on the interaction between unfavorable socioeconomic conditions, younger age of the child, and severity of the diarrhea. PMID- 12764462 TI - [Conceptual basis for the application of biomonitoring on river water quality evaluation programs]. AB - Biomonitoring is defined as the systematic use of biological responses to assess environmental changes, usually anthropogenic impacts. In this article we present the conceptual basis and a brief history of biomonitoring as an assessment tool for environmental health. Considering the drawbacks of physical and chemical parameters to assess environmental quality, we pinpoint the need to integrate these analyses with information provided by biological monitoring. The application of biomonitoring in Brazil would help watershed managers and policy makers to reduce costs, increase the efficiency of analyses, and simplify the results, allowing community participation through volunteer monitoring programs. PMID- 12764463 TI - [Control of lead emissions from a battery repair shop in the city of Rio de Janeiro using the air as indicator]. AB - Lead levels in the air surrounding a battery repair shop (BRS) located in the Olaria neighborhood (Rio de Janeiro, Brazil) were monitored from April to July 1999. Most of the samples collected within 25 meters of the BRS exceeded the limit of 1.5 micro g.Pb.m-3 established by the United States Environmental Protection Agency (EPA). These results were published in a previous article (Quiterio et al., 2001). In the current study, installation of a Venture ventilation system and some changes in the operational procedure were proposed. After the modifications in the BRS, a new monitoring campaign was performed (August and September, 2000). Three points were selected for air collection, corresponding to the most critical points found in the previous assessments. The new concentrations ranged from 0.8 to 17.6 micro g.Pb.m-3, showing that lead emissions into the environment had decreased adequately. However, concentration at the chimney exhaust was still higher than EPA limits, and further improvements in the installations and procedures are definitely needed. PMID- 12764464 TI - [Non-fatal occupational injuries: gender and job contract differences]. AB - In Brazil, workers without a formal job contract comprise more than half of the labor force, a contingent formed mainly by women. This study presents estimates of the annual incidence of non-fatal work-related injuries and their distribution by gender and type of job contract in an urban area of Brazil. This was a community-based study with a random cluster area sample of the inhabitants from the city of Salvador, capital of the State of Bahia. The study population included all individuals from 18 to 65 years of age who reported having a paid job (n = 2,947). Data were obtained through individual household interviews. The overall estimated annual incidence rate was 5.80%, with a non-statistically significant difference between men (6.05%) and women (5.53%) or workers with (5.67%) and without (5.92%) a formal job contract. These similarities between work-related accidents across gender and type of job contract highlighted the need for greater attention to this problem among women and informal workers, who are largely ignored in official statistics. PMID- 12764465 TI - [Shared development of printed material as a health education strategy with public school employees in Rio de Janeiro State]. AB - This article describes the process and results of an experience with preparing a mural entitled A Day in the Life of the School Lunch Cook, conducted with public school employees in the State of Rio de Janeiro by a team of researchers and students from the Specialization Course in Workers' Health at the Center for Studies on Workers' Health of the National School of Public Health, Oswaldo Cruz Foundation, in partnership with the School Employees' Union. "Health workshops" based on participatory research were used to raise questions related to daily work by school lunch cooks and kitchen staff. Produced to share the results of workshops with other schools, the mural is highly meaningful in terms of the reality of a specific segment of public employees and is intended as food for thought. Based on the Paulo Freire popular education model, six meetings were held, following a script for the production of an educational leaflet including the selection of contents, the communications approach, and the style and format of the leaflet. The creativity in the images prepared by the school lunch employees based on actual experiences from their daily work made the workshops an enriching moment of exchange, in which the participants shared their ideas, values, beliefs, feelings, and life histories. PMID- 12764466 TI - [Comparison of nutritional status of low-income children in the second year of life, before and after primary health care program implantation]. AB - The so-called "Slum Project" or Projeto Favela at the Sao Paulo School of Medicine, Federal University in Sao Paulo, conducts numerous primary health care activities in low-income communities in the city of S o Paulo to prevent and control nutritional deficiencies. The aim of this study was to compare the nutritional status of children from 12 to 26 months of age before and after preventive intervention. The study was conducted with two groups, before and after intervention, with 57 and 65 children, respectively. Anthropometric measures and hemoglobin values were obtained. In the before-intervention group, the prevalence of malnutrition as measured by weight for age and weight for height was 12.3% and 1.8% respectively, whereas there were no cases of malnutrition in the after-intervention group. There was a reduction in the prevalence of anemia, especially the severe form, from 33% to 7%. Mean hemoglobin increased from 10.1 to 11.0 g/dl, and the duration of breastfeeding was greater (> 180 days) after intervention. PMID- 12764467 TI - [Driving-related behavior: an epidemiologic study of undergraduate students]. AB - The purpose of this article was to describe self-reported driving-related behavior and compare the frequency of risk-taking among drivers with and without a history of traffic accidents (TA). A cross-sectional study was designed, and 2,116 undergraduate students from a public university in Brazil ranging in age from 18 to 25 were interviewed. Association between independent variables and history of TA was described using frequency, chi2, p, odds ratio, and 95% confidence intervals. Male data were analyzed through logistic regression. Men showed a higher risk than women of having been involved in TA. Drivers with more frequent risk-taking had a greater risk of history of TA. Behaviors observed to be associated with TA in men were: "history of fines", "driving on the shoulder", and "drinking and driving". Open-ended questions demonstrated that students show low awareness of their own responsibility in TA. Interventions among students are necessary and must prioritize males and mainly those from upper socioeconomic groups. PMID- 12764468 TI - [New educational strategies: evaluation of the Jury Trial technique for training council members in women's health in Santa Catarina, Brazil]. AB - This article describes and evaluates the Jury Trial technique, an innovative educational instrument used in a pilot workshop on Social Control and Gender by health council members in Greater Metropolitan Florian polis, Santa Catarina, Brazil. The reliability and validity were studied based on the participants' opinions, complemented by qualitative observation of the work. There was consensus in acceptance of the technique based on "acquisition of knowledge" and the fact that it was participatory, democratic, and dynamic. The positive evaluation agrees with a previous study that interviewed municipal health secretaries, with a smaller preference of formal cognitive activities that were also part of the workshop. The article concludes by emphasizing the importance of this innovative technique in training programs, since it is accepted by different target publics, allows for designing the program contents, and is useful in relation to new and controversial issues since it favors practical and reflexive experiences. PMID- 12764469 TI - [Impact of antiretroviral therapy on the magnitude of the HIV/AIDS epidemic in Brazil: various scenarios]. AB - We applied the back-calculation method to estimate the magnitude of the HIV epidemic in Brazil, using the EM and EMS algorithms. Under certain assumptions regarding the behavior of infected patients towards combined antiretroviral therapy, we discuss five different scenarios applied to the Brazilian epidemic. Our objective was to illustrate the impact of combined antiretroviral treatment on the incubation period and thus on estimates of the size of the HIV-infected population, based on reported AIDS cases. PMID- 12764470 TI - [Nutritional status of indigenous children from the Alto Xingu in 1980 and 1992 and follow-up of weight and height from the first through the fourth years of life]. AB - This study focused on the under-five population of the Alto Xingu region in Brazil, with the following objectives: (1) to evaluate height and weight increment from the first through the fourth years of life and (2) to compare nutritional status in 1980 and 1992. Height and weight increases were evaluated in 81 children. Weight and height were measured in 264 children evaluated in 1980 and in 172 in 1992 (< 10 years of age). Median Z-scores in the first and fourth years of life, respectively, showed: (1) a decrease in weight-for-age, (-0.12 in the first year and -0.51 in the fourth year of life; p = 0.002); (2) a decrease in weight-for-height (+1.31 and +0.08; p < 0.001); (3) an increase in height-for age (-1.50 and -0.94; p < 0.001). Median Z-scores in 1980 and 1992 showed: (1) no change in weight-for-age (-0.61 in 1980 and -0.62 in 1992; p = 0.90); (2) no change in weight-for-height (+0.27 and +0.34; p = 0.10); and (3) a decrease in height-for-age (-1.04 and -1.22; p = 0.02). Height-for-age increased and weight for-height decreased between the first and fourth years of life. A decrease in height-for-age was observed from 1980 to 1992, demonstrating the importance of nutritional surveillance among the population of the Alto Xingu. PMID- 12764471 TI - [Prevalence of near vision deficiency and related factors: a population-based study]. AB - The objective of this article was to estimate the prevalence of presbyopia in adults > or = 30 years and explore associated characteristics such as demographic, socioeconomic, and individual variables. A cross-sectional, population-based study was performed with 3,007 adults > or = 30 years in Pelotas, Rio Grande do Sul, Brazil. Near vision acuity was evaluated using the Jaeger table, and other variables were investigated using standardized, pre-coded questionnaires. Prevalence of presbyopia was 54.7%. When visual acuity was measured using visual corrective means, prevalence was 29.34%. Age and female gender were associated with the outcome variable after controlling for confounding. Some 42.4% of the sample reported not requiring near vision for routine daily tasks. The conclusion was that presbyopia displays a high prevalence in the adult population and is directly related to increasing age, especially 45 years of age and appearing earlier in women. In some 30% of individuals who already use visual corrective means, the latter are ineffective for near vision. PMID- 12764472 TI - Evaluation of informative materials on leishmaniasis distributed in Brazil: criteria and basis for the production and improvement of health education materials. AB - Based on categories related to structure, content, language, and illustrations, the present study provides an evaluation of the quality of educational materials on leishmaniasis available to health services in Brazil. The 18 publications evaluated consisted of four handbooks, four guided studies, four booklets, and six leaflets. Of the total publications assessed, nine were produced by the Brazilian National Health Foundation (FUNASA), five by State and Municipal Health Departments jointly with FUNASA, and one by the Pan-American Health Organization. The evaluations were also performed by three professionals: a physician specialized in leishmaniasis, a parasitologist, and an information/communications expert. The publications failed to specify key items such as target public, objective, and bibliography. The illustrations, especially in the booklets and leaflets, failed to clarify the text, portrayed biased concepts, and omitted credits and scale. According to this study, informative materials on leishmaniasis distributed in Brazil present major limitations which jeopardize the quality of information they contain. PMID- 12764473 TI - [Evaluation of a cervical cancer control program in a local health system in Southeast Brazil]. AB - This study considers the possibility of expanding the roles of the local health care system to respond to demands by the population. Focusing on the problem of cervical cancer, the study evaluated the results obtained by a preventive program conducted in a municipal health system with the objective of evaluating its case resolving capacity and patient compliance. Variables were related to diagnosis and follow-up of 465 women enrolled in the program from 1987 to 1994. For purposes of comparison, the study used the results from a specialized regional referral service during the same period. The program's case-resolving capacity was similar to that of the referral service; in addition, on average the cervical lesions were less severe and the treatment drop-out rate was lower among patients from the local health care system, reflecting better follow-up compliance. It is important to train local health personnel in order to improve care, including treatment for more complex diseases. PMID- 12764474 TI - [Social participation in health services: concepts of users and community leaders in two municipalities of Northeastern Brazil]. AB - Social participation in management of the health care system at different levels of government is one of the key policies promoted by the Unified National Health System (SUS) in Brazil. As with any new policy, success hinges on several factors such as stakeholders' interests and opinions, which have not always been considered in the past. This paper analyzes the underlying concepts of two groups of stakeholders with respect to social participation in health and the potential influence of these concepts on the effectiveness of policy implementation. A case study of two municipalities in Northeast Brazil was conducted using a combination of qualitative and quantitative social science research methods. Health services users and community leaders were interviewed. Various concepts were found in which the participatory approach to health policies was only partially reflected. Likely influences on stakeholders' concepts of social participation in health are the evolution of the broader Brazilian social context and the traditional performance of health services. Particular attention should be paid to stakeholders' opinions and concepts if policy effectiveness is to be improved. PMID- 12764475 TI - [Predictors of compliance with highly active antiretroviral therapy in Brasilia, Distrito Federal, Brazil, 1999-2000]. AB - Non-compliance with highly active antiretroviral therapy can reduce treatment efficacy and lead to viral resistance. Side effects can interfere with patients' quality of life. Our objectives were to estimate levels of treatment compliance and investigate the main predictors of non-compliance. The study design was cross sectional. For purposes of comparison, two different percentage cut-offs for compliance were defined for "proper compliance", namely the adequate ingestion of at least 80% or 95% of the prescribed medicines, respectively. Semi-structured interviews were performed in a sequential sample of 150 patients during out patient visits to the Day Hospital in Central Brasilia. Mean compliance was 85.8%. Variables associated with non-compliance were age, schooling, employment status, monthly personal and family income, illegal substance use, family and community structure, presence of opportunistic infections when HIV was diagnosed, and treatment side effects. Prevalence ratios varied from 1.6 to 4.5. We conclude that social and economic variables, as well as those related to individual habits, were the main predictors of compliance. Few variables related to clinical status or treatment were associated with compliance. PMID- 12764476 TI - Obesity and underweight among Brazilian elderly: the Bambui Health and Aging Study. AB - The coexistence of obesity (body mass index, BMI > or = 30kg/m ) and underweight (BMI <= 20kg/m ) and related factors were investigated among all residents aged 60+ years in Bambu , Minas Gerais State, using multinomial logistic regression. 1,451 (85.5%) of the town's elderly participated. Mean BMI was 25.0 (SD = 4.9kg/m ) and was higher for women and decreased with age. Prevalence of obesity was 12.5% and was positively associated with female gender, family income, hypertension, and diabetes and inversely related to physical activity. Underweight affected 14.8% of participants, increased with age, and was higher among men and low-income families. It was negatively associated with hypertension and diabetes and directly associated with Trypanosoma cruzi infection and > or = 2 hospitalizations in the previous 12 months. Both obesity and underweight were associated with increased morbidity. The association of underweight with T. cruzi infection, increased hospitalization, and low family income may reflect illness related weight loss and social deprivation of elderly in this community. Aging in poverty may lead to an increase in nutritional deficiencies and health-related problems among the elderly. PMID- 12764477 TI - [Regional variations in homicide mortality in Jalisco, Mexico]. AB - The present study describes regional variations in homicide rates in Jalisco State, Mexico, in 1989-1991, 1994-1996, and 1999-2000, analyzing the trends by gender and socioeconomic stratum. Using mortality data generated by the National Institute for Statistics, Geography, and Information Technology, homicide rates adjusted by age and gender were calculated, along with rate/female rate ratios; rate ratios by socioeconomic stratum and 95% confidence intervals were also calculated. According to the results, the homicide rate showed: a downward trend in the 1990s; a regional homicide mortality pattern, with the highest rates in peripheral regions, considered among the poorest areas in the State; municipalities with the lowest socioeconomic conditions also presenting a statistically significant excess homicide mortality; and an evident over mortality from homicide among males. The results point to tasks and challenges for public health and law enforcement institutions, including the need to implement different inter-institutional policies that take into consideration the characteristics of homicide and violent crime in Jalisco. PMID- 12764478 TI - [Social support: scale test-retest reliability in the Pro-Health Study]. AB - This article describes the test-retest reliability of a scale comprising five dimensions of social support: material, emotional, informational, affective, and positive social interaction. In the study, a sample of 192 employees at a university in Rio de Janeiro filled out the same questionnaire on two occasions, 15 days apart. Measures of stability used were the intraclass correlation coefficient (ICC), weighted kappa statistic, and log-linear models. Internal consistency was evaluated using the Cronbach's alpha coefficient. Social support dimensions showed internal consistency ranging from 0.75 to 0.91 at test, and 0.86 to 0.93 at retest. The ICC ranged from 0.78 to 0.87 in the five dimensions of the scale, with no substantial differences by gender, age, or level of schooling. For most questions, the "agreement plus linear by linear association" and "quasi-association" log-linear models gave the best fit. According to these results, the reliability of the instrument was considered adequate, enabling it to be used in ongoing assessment of associations between social support and health-related outcomes in a cohort study (the Pro-Health Study) recently begun in Rio de Janeiro. PMID- 12764479 TI - [Western medicine and alternative medicines: can they be complementary? Conceptual reflections]. AB - The present article is part of a series of reflections from an intercultural approach to health systems and corresponding public policies, motivated by findings from a study on two intercultural hospitals in rural Mexico. The frequent utilization of complementary and alternative medicines by the local population and the hegemonic health model that excludes them make the existing health system an unsatisfactory response to people's needs. We present the concept of complementariness as a health system component and propose priorities on this issue, taking different approaches: complementariness as a public policy, as an institutional project, or as an individual decision by the therapist or patient. PMID- 12764480 TI - [Death by AIDS or maternal death: classification of mortality as a social practice]. AB - This paper analyzes the decision on whether to include deaths of HIV+ pregnant women in the classification of maternal mortality. The study focuses on deaths of childbearing-age women in S o Paulo in 1998, investigated by the State's Central Committee on Maternal Mortality (CCMM). Working from a social constructionist perspective, the research was based on documental analysis and interviews with the president of the CCMM and members of one regional committee. The analysis focused on the selection, investigation, and classification of maternal death, with special attention to the negotiation among various actors involved in the classificatory procedures. The data suggest that in the deaths of HIV+ women, other factors are present including the precedence of HIV status over maternal death and the moral and symbolic aspects of AIDS. The results suggest that improving data through a better understanding of decisions to include or exclude cases is only one side of the issue; it is also important to improve quality of care in pregnancy and childbirth in order to prevent maternal death. PMID- 12764481 TI - [Collective health, the new genetics, and market eugenics]. AB - Due to constant advances in genetic manipulation, the field of public health is faced with the possible emergence of a reductionist genomic emphasis, beyond socio-cultural influences. Under such circumstances, the main focus of public health interventions would shift towards a genomic approach to "individuals" and their "families", to the detriment of their consecrated object - "populations", thereby moving away from the current central priority of efforts (and resources) aimed at reducing ill-health due to socioeconomic inequalities. Even admitting the benefits of genetic knowledge, a new eugenic practice may emerge from the availability of genetic tests on the marketplace aimed at individuals that can afford to consume them. PMID- 12764482 TI - [Cooperation between an NGO and "host" states in the control of leprosy in Latin America]. AB - The proliferation of nongovernmental organizations (NGOs) can be considered the result of the inability of the current democratic system to perform all the tasks desired by its citizens. Although NGOs often do quite positive work, they tend to diminish governmental power and are capable of interfering in the internal affairs of other countries. In this context, there are efforts to control their activities, and this control can produce both negative effects (blocking the defense of human rights) and positive ones (correcting the lack of coordination in the work by NGOs). NGOs working with the control of leprosy have a long history of cooperation with "host" states in Latin America. In the worst cases they work in a vacuum left by the state. In a country like Brazil, where the government prioritizes the control of Hansen disease and community participation in the political process - NGOs generally work "in harmony" with national authorities. The most useful contribution to state efforts has been the technical and financial support for training health personnel, supervision, and awareness raising campaigns. Thus, the NGO becomes "quasi-governmental" in performing its tasks. PMID- 12764483 TI - [Intestinal parasite infections in a semiarid area of Northeast Brazil: preliminary findings differ from expected prevalence rates]. AB - We report on intestinal parasite infection prevalence in a population sample from S o Raimundo Nonato, Southeast Piau State, Brazil, aimed at comparison with previous studies on Trichuris trichiura and Ascaris lumbricoides infection. A total of 265 stool specimens were collected and examined by spontaneous sedimentation. Approximately 57% of specimens were infected with at least one parasite species. Entamoeba coli (35.8%), Endolimax nana (13.6%), Hymenolepis nana (9.4%), and hookworm (9.4%) were the most frequently observed parasites. Two cases of roundworm infection were detected, probably acquired outside the region. T. trichiura eggs were not found. Interestingly, neither A. lumbricoides nor T. trichiura has been found in local prehistoric human coprolites. Nevertheless, hookworm infection has been present in the region for at least 7,000 years. PMID- 12764484 TI - [Polycyclic aromatic hydrocarbons in industrial solid waste: a preliminary evaluation of the potential risk of environmental and human contamination in waste disposal areas]. AB - Proper solid waste disposal is important to avoid human and environmental contamination. The NBR 10,004 Waste Classification lists several polycyclic aromatic hydrocarbons (PAHs) and indicates that the presence of at least one PAH in a waste sample is enough to classify it as hazardous. The aim of this study was a preliminary evaluation of PAHs in solid waste samples from selected industries to obtain a preliminary overview of their potential for contamination in case of improper disposal. One or more PAHs listed in NBR 10,004 (benzo[a]anthracene, benzo[a]pyrene, benzo[b]fluoranthene, benzo[k]fluoranthene, indene[1,2,3-c,d]pyrene, chrysene, or fluoranthene) were found in all samples, thus leading to their classification as hazardous waste. Our results showed that toxicologically relevant PAHs were found in all the samples, indicating that their final disposal must be performed in appropriate areas in order to minimize human health risks and environmental contamination from waste disposal areas. PMID- 12764485 TI - [Risk factors for Giardia lamblia infection in children in daycare centers in Havana, Cuba]. AB - We conducted a longitudinal study on giardiasis in three daycare centers in Havana City for a period of 18 months and described a group of children with a "predisposition" or tendency towards re-infection with Giardia lamblia. This group was found to be more frequently associated with clinical symptoms such as diarrhea. A case-control study was designed to determine whether socioeconomic factors and hygiene were associated with this phenomenon. We found no differences between the groups with regard to overcrowding rates, number of persons per bed, absence of certain electric appliances, mother's schooling, or mean family income. However, there were proportionally more fathers with less than complete secondary education among cases as compared to controls. In addition, we found no differences in lack of hand-washing before eating and after defecation; however, we found a higher percentage of families who washed vegetables insufficiently among predisposed children. Finally, a lower percentage of families with predisposed children boiled their water. Our results demonstrate the important role of water as a vehicle for transmission of giardiasis and the importance of various epidemiologic factors. PMID- 12764486 TI - [Evaluation of noise pollution in the Botanical Garden in Curitiba, Parana, Brazil]. AB - This study focuses on noise pollution in the Botanical Garden in Curitiba, Parana, Brazil. Equivalent noise levels (Leq) were measured at 21 points throughout the park, and interviews were conducted with park visitors. Some 47.6% of the measurement sites presented Leq levels over 65dB(A), considered by preventive medicine as the maximum tolerable exposure level without risk of health impairment, and 90.5% of the sites failed to comply with Municipal Ordinance 8,583, setting 55dB(A) as the maximum noise emission level for green areas. The results of interviews with visitors showed that 78% visit the park at least twice a week and that 96% come for physical activity. During their activities in the Botanical Garden, 24% of interviewees identified noise pollution as a source of annoyance, as compared to 22% who complained of insufficient park security. PMID- 12764487 TI - Public-private partnerships for health require thoughtful evaluation. PMID- 12764488 TI - The global burden of diarrhoeal disease in children. PMID- 12764489 TI - Implementation of WHO guidelines on management of severe malnutrition in hospitals in Africa. AB - OBJECTIVE: To investigate the problems, benefits, feasibility, and sustainability of implementation of WHO guidelines on management of severe malnutrition. METHODS: A postal survey invited staff from 12 African hospitals to participate in the study. Five hospitals were evaluated and two were selected to take part in the study: a district hospital in South Africa and a mission hospital in Ghana. At an initial visit, an experienced paediatrician reviewed the situation in the hospitals and introduced the principles of the guidelines through a participatory approach. During a second visit about six months later, the paediatrician reviewed the feasibility and sustainability of the introduced changes and helped find solutions to problems. At a final visit after one year, the paediatrician reassessed the overall situation. FINDINGS: Malnutrition management practices improved at both hospitals. Measures against hypoglycaemia, hypothermia, and infection were strengthened. Early, frequent feeding was established as a routine practice. Some micronutrients for inclusion in the diet were not locally available and needed to be imported. Problems were encountered with monitoring of weight gain and introducing a rehydration solution for malnutrition. CONCLUSION: Implementation of the main principles of the WHO guidelines on severe malnutrition was feasible, affordable, and sustainable at two African hospitals. The guidelines could be improved by including suggestions on how to adapt specific recommendations to local situations. The guidelines are well supported by experience and published reports, but more information is needed about some components and their impact on mortality. PMID- 12764490 TI - Non-specific effect of measles vaccination on overall child mortality in an area of rural India with high vaccination coverage: a population-based case-control study. AB - OBJECTIVE: To determine whether vaccination against measles in a population with sustained high vaccination coverage and relatively low child mortality reduces overall child mortality. METHODS: In April and May 2000, a population-based, case control study was conducted at Ballabgarh (an area in rural northern India). Eligible cases were 330 children born between 1 January 1991 and 31 December 1998 who died aged 12-59 months. A programme was used to match 320 controls for age, sex, family size, and area of residence from a birth cohort of 15 578 born during the same time period. FINDINGS: The analysis used 318 matched pairs and suggested that children aged 12-59 months who did not receive measles vaccination in infancy were three times more likely to die than those vaccinated against measles. Children from lower caste households who were not vaccinated in infancy had the highest risk of mortality (odds ratio, 8.9). A 27% increase in child mortality was attributable to failure to vaccinate against measles in the study population. CONCLUSION: Measles vaccine seems to have a non-specific reducing effect on overall child mortality in this population. If true, children in lower castes may reap the greatest gains in survival. The findings should be interpreted with caution because the nutritional status of the children was not recorded and may be a residual confounder. "All-cause mortality" is a potentially useful epidemiological endpoint for future vaccine trials. PMID- 12764491 TI - Transgenic mice as an alternative to monkeys for neurovirulence testing of live oral poliovirus vaccine: validation by a WHO collaborative study. AB - OBJECTIVE: Extensive WHO collaborative studies were performed to evaluate the suitability of transgenic mice susceptible to poliovirus (TgPVR mice, strain 21, bred and provided by the Central Institute for Experimental Animals, Japan) as an alternative to monkeys in the neurovirulence test (NVT) of oral poliovirus vaccine (OPV). METHODS: Nine laboratories participated in the collaborative study on testing neurovirulence of 94 preparations of OPV and vaccine derivatives of all three serotypes in TgPVR21 mice. FINDINGS: Statistical analysis of the data demonstrated that the TgPVR21 mouse NVT was of comparable sensitivity and reproducibility to the conventional WHO NVT in simians. A statistical model for acceptance/rejection of OPV lots in the mouse test was developed, validated, and shown to be suitable for all three vaccine types. The assessment of the transgenic mouse NVT is based on clinical evaluation of paralysed mice. Unlike the monkey NVT, histological examination of central nervous system tissue of each mouse offered no advantage over careful and detailed clinical observation. CONCLUSIONS: Based on data from the collaborative studies the WHO Expert Committee for Biological Standardization approved the mouse NVT as an alternative to the monkey test for all three OPV types and defined a standard implementation process for laboratories that wish to use the test. This represents the first successful introduction of transgenic animals into control of biologicals. PMID- 12764492 TI - Self-treatment of malaria in rural communities, Butajira, southern Ethiopia. AB - OBJECTIVES: To quantify the use of self-treatment and to determine the actions taken to manage malaria illness. METHODS: A cross-sectional study was undertaken in six peasant associations in Butajira district, southern Ethiopia, between January and September 1999. Simple random sampling was used to select a sample of 630 households with malaria cases within the last six months. FINDINGS: Overall, 616 (>97%) of the study households acted to manage malaria, including the use of antimalarial drugs at home (112, 17.8%), visiting health services after taking medication at home (294, 46.7%), and taking malaria patients to health care facilities without home treatment (210, 33.3%). Although 406 (64.5%) of the households initiated treatment at home, the use of modern drugs was higher (579, 92%) than that of traditional medicine (51, 8%). Modern drugs used included chloroquine (457, 73.5%) and sulfadoxine-pyrimethamine (377, 60.6%). Malaria control programmes were the main sources of antimalarials. In most cases of malaria, treatment was started (322, 52.3%) or health services visited (175, 34.7%) within two days of the onset of symptoms. Cases of malaria in the lowland areas started treatment and visited health services longer after the onset of malaria than those in the midland areas (adjusted odds ratio, 0.44; 95% confidence interval (CI), 0.30-0.64; and adjusted odds ratio, 0.37; 95% CI, 0.25 0.56, respectively). Similarly, those further than one hour's walk from the nearest health care facility initiated treatment later than those with less than one hour's walk (adjusted odds ratio, 0.62; 95% CI 0.43-0.87). This might be because of inaccessibility to antimalarial drugs and distant health care facilities in the lowland areas; however, statistically insignificant associations were found for sex, age, and religion. CONCLUSION: Self-treatment at home is the major action taken to manage malaria. Efforts should be made to improve the availability of effective antimalarials to communities in rural areas with malaria, particularly through the use of community health workers, mother coordinators, drug sellers, and shop owners. PMID- 12764493 TI - Cost-effectiveness of social marketing of insecticide-treated nets for malaria control in the United Republic of Tanzania. AB - OBJECTIVE: To assess the costs and consequences of a social marketing approach to malaria control in children by means of insecticide-treated nets in two rural districts of the United Republic of Tanzania, compared with no net use. METHODS: Project cost data were collected prospectively from accounting records. Community effectiveness was estimated on the basis of a nested case-control study and a cross-sectional cluster sample survey. FINDINGS: The social marketing approach to the distribution of insecticide-treated nets was estimated to cost 1560 US dollars per death averted and 57 US dollars per disability-adjusted life year averted. These figures fell to 1018 US dollars and 37 US dollars, respectively, when the costs and consequences of untreated nets were taken into account. CONCLUSION: The social marketing of insecticide-treated nets is an attractive intervention for preventing childhood deaths from malaria. PMID- 12764494 TI - The cost-effectiveness of policies for the safe and appropriate use of injection in healthcare settings. AB - OBJECTIVE: Poor injection practices transmit potentially life-threatening pathogens. We modelled the cost-effectiveness of policies for the safe and appropriate use of injections in ten epidemiological subregions of the world in terms of cost per disability-adjusted life year (DALY) averted. METHODS: The incidence of injection-associated hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency virus (HIV) infections was modelled for a year 2000 cohort over a 30-year time horizon. The consequences of a "do nothing" scenario were compared with a set of hypothetical scenarios that incorporated the health gains of effective interventions. Resources needed to implement effective interventions were costed for each subregion and expressed in international dollars (I dollars). FINDINGS: Worldwide, the reuse of injection equipment in the year 2000 accounted for 32%, 40%, and 5% of new HBV, HCV and HIV infections, respectively, leading to a burden of 9.18 million DALYs between 2000 and 2030. Interventions implemented in the year 2000 for the safe (provision of single-use syringes, assumed effectiveness 95%) and appropriate (patients-providers interactional group discussions, assumed effectiveness 30%) use of injections could reduce the burden of injection-associated infections by as much as 96.5% (8.86 million DALYs) for an average yearly cost of 905 million I dollars (average cost per DALY averted, 102; range by region, 14-2293). Attributable fractions and the number of syringes and needles required represented the key sources of uncertainty. CONCLUSION: In all subregions studied, each DALY averted through policies for the safe and appropriate use of injections costs considerably less than one year of average per capita income, which makes such policies a sound investment for health care. PMID- 12764495 TI - International survey on variations in practice of the management of the third stage of labour. AB - OBJECTIVE: To determine the use of the active management of the third stage of labour in 15 university-based obstetric centres in ten developing and developed countries and to determine whether evidence-based practices were being used. METHODS: From March 1999 to December 1999, the Global Network for Perinatal and Reproductive Health (GNPRH) conducted an observational, cross-sectional survey to assess the use of the practice and its components. Prospective data on patient characteristics and the interventions used in the management of the third stage of labour were collected using standardized methods. Data on approximately 30 consecutive vaginal deliveries in each centre (452 in total) were included. FINDINGS: Significant intracountry and intercountry variation in the practice of the active management of the third stage of labour was found (111/452 deliveries used active management), which confirmed the existence of a large gap between knowledge and practice. CONCLUSION: Areas identified for improvement are the urgent implementation of the evidence-based clinical management practice defined as the active management of the third stage of labour; increased accessibility to systematic reviews in developing countries; and the conduction of clinical trials that assess the impact of this intervention in other settings. PMID- 12764496 TI - A new face for private providers in developing countries: what implications for public health? AB - The use of private health care providers in low- and middle-income countries (LMICs) is widespread and is the subject of considerable debate. We review here a new model of private primary care provision emerging in South Africa, in which commercial companies provide standardized primary care services at relatively low cost. The structure and operation of one such company is described, and features of service delivery are compared with the most probable alternatives: a private general practitioner or a public sector clinic. In a case study of cost and quality of services, the clinics were popular with service users and run at a cost per visit comparable to public sector primary care clinics. However, their current role in tackling important public health problems was limited. The implications for public health policy of the emergence of this new model of private provider are discussed. It is argued that encouraging the use of such clinics by those who can afford to pay for them might not help to improve care available for the poorest population groups, which are an important priority for the government. Encouraging such providers to compete for government funding could, however, be desirable if the range of services presently offered, and those able to access them, could be broadened. However, the constraints to implementing such a system successfully are notable, and these are acknowledged. Even without such contractual arrangements, these companies provide an important lesson to the public sector that acceptability of services to users and low-cost service delivery are not incompatible objectives. PMID- 12764497 TI - Standard case management of pneumonia in children in developing countries: the cornerstone of the acute respiratory infection programme. PMID- 12764498 TI - Acute lower respiratory tract infections in children: possible criteria for selection of patients for antibiotic therapy and hospital admission. 1984. PMID- 12764499 TI - Researchers argue that unsafe injections spread HIV more than unsafe sex. PMID- 12764501 TI - Scientific publishers consider censoring "dangerous" research. PMID- 12764500 TI - No deal in sight on cheap drugs for poor countries. PMID- 12764502 TI - Legal interest expands from tobacco to obesity. PMID- 12764503 TI - Assisted suicide seekers turn to Switzerland. PMID- 12764504 TI - Unsafe sex is the main mode of HIV transmission. PMID- 12764508 TI - Healthy environments for children. PMID- 12764509 TI - New water forum will repeat old message. PMID- 12764510 TI - Health research systems: a framework for the future. PMID- 12764511 TI - Evaluation of a water, sanitation, and hygiene education intervention on diarrhoea in northern Pakistan. AB - OBJECTIVE: Inadequate water and sanitation services adversely affect the health and socioeconomic development of communities. The Water and Sanitation Extension Programme (WASEP) project, undertaken in selected villages in northern Pakistan between 1997 and 2001, was designed to deliver an integrated package of activities to improve potable water supply at village and household levels, sanitation facilities and their use, and awareness and practices about hygiene behaviour. METHODS: A case-control study was conducted during July-September 2001 to evaluate whether, after selected confounders were controlled for, children aged <6 years with diarrhoea were more or less likely to reside in villages that participated in the project than in villages that did not participate. Descriptive and logistic regression analyses were performed. FINDINGS: Children not living in WASEP villages had a 33% higher adjusted odds ratio for having diarrhoea than children living in WASEP villages (adjusted odds ratio, 1.331; P<0.049). Boys had 25% lower odds of having diarrhoea than girls (adjusted odds ratio, 0.748; P<0.049). A 2.6% decrease was found in the odds of diarrhoea for every yearly increase in the mother's age (adjusted odds ratio, 0.974; P<0.044) and a 1.4% decrease for every monthly increase in the child's age (adjusted odds ratio, 0.986; P<0.001). CONCLUSIONS: The findings in this study may help refine the approach to future water, sanitation, and hygiene initiatives in northern Pakistan. The integrated approach taken by WASEP, which incorporates engineering solutions with appropriate education to maximize facility usage and improve hygiene practices, is a useful example of how desired health benefits can be obtained from projects of this type. PMID- 12764512 TI - Hypothetical performance of syndrome-based management of acute paediatric admissions of children aged more than 60 days in a Kenyan district hospital. AB - OBJECTIVE: To investigate whether the outpatient, syndrome-based approach of the Integrated Management of Childhood Illness (IMCI) protocol could be extended to the inpatient arena to give clear and simple minimum standards of care for poorly resourced facilities. METHODS: A prospective, one-year admission cohort retrospectively compared hypothetical performance of syndrome-based management with paediatrician-defined final diagnosis. Admission syndrome definitions were based on local adaptations to the IMCI protocol that encompassed 20 clinical features, measurement of oxygen saturation, and malaria microscopy. FINDINGS: After 315 children with clinically obvious diagnoses (e.g. sickle cell disease and burns) were excluded, 3705 admission episodes were studied. Of these, 2334 (63%) met criteria for at least one severe syndrome (mortality 8% vs <1% for "non severe" cases), and half of these had features of two or more severe syndromes. No cases of measles were seen. Syndrome-based treatment would have been appropriate (sensitivity >95%) for severe pneumonia, severe malaria, and diarrhoea with severe dehydration, and probably for severe malnutrition (sensitivity 71%). Syndrome-directed treatment suggested the use of broad spectrum antibiotics in 75/133 (56% sensitivity) children with bacteraemic and 63/71 (89% sensitivity) children with meningitis. CONCLUSIONS: Twenty clinical features, oxygen saturation measurements, and results of malaria blood slides could be used for inpatient, syndrome-based management of acute paediatric admissions. The addition of microscopy of the cerebrospinal fluid and haemoglobin measurements would improve syndrome-directed treatment considerably. This approach might rationalize admission policy and standardize inpatient paediatric care in resource-poor countries, although the clinical detection of bacteraemia remains a problem. PMID- 12764513 TI - Implementation and quantitative evaluation of chronic disease self-management programme in Shanghai, China: randomized controlled trial. AB - OBJECTIVE: To evaluate the effectiveness of the Shanghai Chronic Disease Self Management Program (CDSMP). METHODS: A randomized controlled trial with six-month follow-up compared patients who received treatment with those who did not receive treatment (waiting-list controls) in five urban communities in Shanghai, China. Participants in the treatment group received education from a lay-led CDSMP course and one copy of a help book immediately; those in the control group received the same education and book six months later. FINDINGS: In total, 954 volunteer patients with a medical record that confirmed a diagnosis of hypertension, heart disease, chronic lung disease, arthritis, stroke, or diabetes who lived in communities were assigned randomly to treatment (n = 526) and control (n = 428) groups. Overall, 430 (81.7%) and 349 (81.5%) patients in the treatment and control groups completed the six-month study. Patients who received treatment had significant improvements in weekly minutes of aerobic exercise, practice of cognitive symptom management, self-efficacy to manage own symptoms, and self-efficacy to manage own disease in general compared with controls. They also had significant improvements in eight indices of health status and, on average, fewer hospitalizations. CONCLUSION: When implemented in Shanghai, the CDSMP was acceptable culturally to Chinese patients. The programme improved participants' health behaviour, self-efficacy, and health status and reduced the number of hospitalizations six months after the course. The locally based delivery model was integrated into the routine of community government organizations and community health services. Chinese lay leaders taught the CDSMP courses as successfully as professionals. PMID- 12764514 TI - Management of acute respiratory infections by community health volunteers: experience of Bangladesh Rural Advancement Committee (BRAC). AB - OBJECTIVE: To assess the role of management practices for acute respiratory infections (ARIs) in improving the competency of community health volunteers in diagnosing and treating acute respiratory infections among children. METHODS: Data were collected by a group of research physicians who observed the performance of a sample of 120 health volunteers in 10 sub-districts in Bangladesh in which Bangladesh Rural Advancement Committee (BRAC) had run a community-based ARI control programme since mid-1992. Standardized tests were conducted until the 95% interphysician reliability on the observation of clinical examination was achieved. FINDINGS: The sensitivity, specificity, and overall agreement rates in diagnosing and treating ARIs were significantly higher among the health volunteers who had basic training and were supervised routinely than among those who had not. CONCLUSION: Diagnosis and treatment of ARIs at the household level in developing countries are possible if intensive basic training and the close supervision of service providers are ensured. PMID- 12764515 TI - Efficacy of oxamniquine and praziquantel in the treatment of Schistosoma mansoni infection: a controlled trial. AB - OBJECTIVE: To evaluate the therapeutic efficacy of oxamniquine and praziquantel, the two most clinically important schistosomicide drugs, and to compare the accuracy of faecal examination with the accuracy of oogram in testing for Schistosoma mansoni infection. METHODS: In a triple-masked and randomized controlled trial, 106 patients infected with S. mansoni were randomly allocated to one of three statistically homogeneous groups. One group was given 60 mg/kg praziquantel per day for three consecutive days, another was given two daily doses of 10 mg/kg oxamniquine, and the placebo group received starch. Faecal examinations (days 15, 30, 60, 90, 120, 150, and 180 after treatment) and biopsy of rectal mucosa by quantitative oogram (days 30, 60, 120, and 180) were used for the initial diagnosis and for evaluating the degree of cure. The chi2 test and the Kruskal-Wallis test were used to compare variables in the three groups. Survival analysis (Kaplan-Meier) and the log-rank test were used to evaluate the efficacy of the treatments. FINDINGS: The sensitivity of stool examinations ranged from 88.9% to 94.4% when patients presented with >5000 S. mansoni eggs per gram of tissue (oogram); when the number of eggs dropped to <1000 eggs per gram, sensitivity was reduced (range, 22.7-34.0%). When cure was evaluated by stool examination, oxamniquine and praziquantel had cure rates of 90.3% and 100%, respectively. However, when the oogram was used as an indicator of sensitivity, the oxamniquine cure rate dropped dramatically (to 42.4%), whereas the rate for praziquantel remained high, at 96.1%. CONCLUSIONS: Praziquantel was significantly more effective than oxamniquine in treating S. mansoni infection. The oogram was markedly more sensitive than stool examinations in detecting S. mansoni eggs and should be recommended for use in clinical trials with schistosomicides. PMID- 12764516 TI - The global burden of diarrhoeal disease, as estimated from studies published between 1992 and 2000. AB - Current estimates of the global burden of disease for diarrhoea are reported and compared with previous estimates made using data collected in 1954-79 and 1980 89. A structured literature review was used to identify studies that characterized morbidity rates by prospective surveillance of stable populations and studies that characterized mortality attributable to diarrhoea through active surveillance. For children under 5 years of age in developing areas and countries, there was a median of 3.2 episodes of diarrhoea per child-year. This indicated little change from previously described incidences. Estimates of mortality revealed that 4.9 children per 1000 per year in these areas and countries died as a result of diarrhoeal illness in the first 5 years of life, a decline from the previous estimates of 13.6 and 5.6 per 1000 per year. The decrease was most pronounced in children aged under 1 year. Despite improving trends in mortality rates, diarrhoea accounted for a median of 21% of all deaths of children aged under 5 years in these areas and countries, being responsible for 2.5 million deaths per year. There has not been a concurrent decrease in morbidity rates attributable to diarrhoea. As population growth is focused in the poorest areas, the total morbidity component of the disease burden is greater than previously. PMID- 12764517 TI - Analysis of the effects of malaria chemoprophylaxis in children on haematological responses, morbidity and mortality. AB - This paper reviews the evidence for beneficial effects of malaria chemoprophylaxis on haematological responses, morbidity, mortality, health service utilization and rebound immunity in children. As anaemia may play an important role in childhood mortality, it is important to assess evidence from controlled trials of the potential of chemoprophylaxis to reduce childhood anaemia. An analysis of trials found good evidence that malaria chemoprophylaxis improves mean haemoglobin levels and reduces severe anaemia, clinical malaria attacks, parasite and spleen rates. Significant reductions in outpatient attendance and hospital admissions have been achieved, and substantial evidence from Gambian studies shows reductions in mortality. Chemoprophylaxis in children does not seem to produce any sustained impairment of immunity to malaria, although rebound effects may be greater in children who receive prophylaxis during infancy. Short periods of targeted prophylaxis are likely to be preferable to continuous drug administration. Evidence of the protective efficacy of malaria chemoprophylaxis in children shows that this strategy could be considered within integrated health programmes for specific time periods. Intermittent routine combination therapy early in childhood may be appropriate for those living under holoendemic conditions. Large-scale studies over a number of years are needed to address this issue and the impact of this approach on health service utilization, mortality, and the emergence of drug-resistant parasites. PMID- 12764518 TI - The Singapore Tuberculosis Elimination Programme: the first five years. AB - The Singapore Tuberculosis Elimination Programme (STEP) was launched in 1997 because the incidence of the disease had remained between 49 and 56 per 100,000 resident population for the preceding 10 years. STEP involves the following key interventions: directly observed therapy (DOT) in public primary health care clinics; monitoring of treatment progress and outcome for all cases by means of a National Treatment Surveillance Registry; and preventive therapy for recently infected close contacts of infectious tuberculosis cases. Among other activities are the revamping of the National Tuberculosis Notification Registry, the discontinuation of BCG revaccination for schoolchildren, the tightening up of defaulter tracing, and the education of the medical community and the public. Future plans include an outreach programme for specific groups of patients who are unable to attend their nearest public primary care clinics for DOT, the detention of infectious recalcitrant defaulters for treatment under the Infectious Diseases Act, the molecular fingerprinting of tuberculosis isolates, and targeted screening of high-risk groups. The incidence of tuberculosis fell from 57 per 100,000 population in 1998 to 48 per 100,000 in 1999 and continued to decline to 44 per 100,000 in 2001. With political will and commitment and the support of the medical community and the public it is hoped that STEP will achieve further progress towards the elimination of tuberculosis in Singapore. PMID- 12764519 TI - The Healthy Cities approach-- reflections on a framework for improving global health. PMID- 12764520 TI - Advancing the state of the world's newborns. PMID- 12764521 TI - Can international public health law help to prevent war? PMID- 12764522 TI - Two new reports conclude that the health costs of war are unaffordable. PMID- 12764523 TI - Brazil mobilizes for a war to save lives. PMID- 12764524 TI - Health standards plummet on occupied farms in Zimbabwe. PMID- 12764530 TI - Extension of organ transplantation: some ethical considerations. AB - The concept of vital organ transplantation is critically analyzed by considering how traditional transplantation modifies the commitment to saving lives. Problems such as those associated with immunosuppression might seem to provide a compelling reason to oppose extension of transplantation to non-lifesaving situations. A closer examination, however, shows that immunosuppression does not present an intractable objection. For some organ transplants, such as the uterus, use of immunosuppression could be limited to the childbearing years. Complexities associated with assessment of quality of life are discussed using the example of hand transplantation. Assessment of success and functionality in a hand transplant is more complicated than it might appear at first. These complications suggest that monitoring and assessment should be a part of any extension of organ transplantation. Informed consent provides a limited, but important, component in justifying extended transplantation. Such justification, however, does not rest on patient autonomy, but on the reasonable prospect of benefit. Transplant programs considering an extension of traditional transplantation should develop formal protocols that include assessment of costs, benefits, quality of life, and the adequacy of informed consent. PMID- 12764531 TI - Hand transplantation: current status. AB - The hand is a very special organ, with unique functions and versatility in the human body. Our hands are pivotal in manipulating our environment, receiving feedback from our surroundings and communicating our unspoken words by gestures. Thus, the loss of a hand is a tragic, disfiguring event with profound personal, vocational, financial and social implications. Transplantation of life-saving solid organs is now widely accepted in both the medical and lay communities. The technical skills and prerequisites for hand transplantation have been honed over recent decades, culminating in the recent commencement of hand transplantation in several centers around the world. However, unlike life-saving solid organ transplantation, hand transplantation has been greeted with less enthusiasm in the professional community because it is not yet clear what the long-term risks to-benefits ratio is. The scientific background, and the potential risks, benefits, and ethical aspects of this procedure are discussed. Successful transplantation to amputees of fully integrated and functional hands is a worthy goal. Hopefully, at some point in the future, hand transplantation will become another safe and viable option for amputees to consider. PMID- 12764532 TI - Uterus transplantation. AB - Until recently, only life and death situations warranted organ transplantation. Nonvital transplantation, to further a patient s wishes and goals, was not considered justified. It can be argued, however, that this distinction is not morally significant. Patients with kidney failure, for example, can be kept alive by dialysis. But their quality of life would be greatly enhanced by kidney transplant, which is thus considered a justified procedure. So a spectrum of rationales may justify transplantation. Transplantation of the uterus would relieve the anguish of women who greatly desire to conceive a child. Some women do not have a uterus. In some cases this is due to a congenital absence (Rokitansky s syndrome). In other cases, surgical removal of the uterus was required to repair an obstetrical rupture. With a transplanted uterus, many of these women could have the opportunity to become pregnant as a result of nonvital organ transplant. While other organ transplant donations most often come from cadavers and less often from living donors (kidney or partial liver), the donor source for a uterus may be an otherwise healthy living patient who requires uterus removal as a standard care procedure. Furthermore, it should be possible to remove the transplanted uterus from the recipient after successful pregnancies, so the patient would not be subjected to lifelong antirejection medications. Since animal uterus transplantation has been done successfully, human uterus transplantation might be considered for select cases. One such case has been reported. PMID- 12764533 TI - Laryngeal and tracheal transplantation: ethical limitations. AB - Over the last decade, there have been extraordinary developments in the field of transplantation science. As a result, organ transplantation enjoys a success that is unparalleled since its introduction nearly 50 years ago. Progress in the laboratory has translated into less toxic, more effective immunosuppressive therapies that have improved both allograft survival and patient quality of life. Consequently, physicians and their patients look toward a new frontier, the transplantation of non-vital organs. While the transplantation of non-vital organs is technically feasible, as demonstrated by the recent success of a human laryngeal transplant, a variety of ethical concerns must be confronted before tracheal and laryngeal transplantation can be offered to patients as a reconstructive option. When considering the risks and benefits of non-vital organ transplantation, one must consider the immeasurable impact of a procedure on the patient s quality of life. The focus of this article is on quality of life and the role of laryngotracheal transplantation in contemporary medicine. PMID- 12764534 TI - Stem cell transplantation. AB - Historically, a variety of hematological disorders of the bone marrow, such as acute leukemia, chronic myelocytic leukemia and severe aplastic anemia, were invariably fatal. In the past two decades, basic and clinical scientists have been able to change this grim reality into a more hopeful outcome by replacing the diseased bone marrow with stem cells from either a family donor or an unrelated volunteer donor with identical human leukocyte antigens (HLA). Because of progress in this field, the initial experience of mainly using sibling donors has now been expanded to the use of either unrelated adult donors or cord blood cells as a source of hematopoietic cells. These approaches have created ethical concerns for donors and families, concerns that need to be discussed and understood by patients, volunteer donors and health care professionals. PMID- 12764535 TI - Candidate selection criteria for living donor liver transplantation. AB - Living donor liver transplantation is a new surgical technique that challenges the assumptions of the current system for allocation of livers for transplantation. Arguments based on justice in the setting of scarcity, and the consequent need for triage, do not apply. The risk to the living donor, an order of magnitude higher than that for kidney donation, raises different ethical concerns related to autonomy, what constitutes an acceptable risk-benefit equation, and who is to decide. PMID- 12764536 TI - Trust and trustworthiness in organ transplantation: good samaritan and emotionally related living donors. AB - Organ transplantation, and living donor organ transplantation in particular, could not occur without society s trust in medicine and in transplant programs. This paper explains the importance of that trust and shows how the concept can be used to define the moral requirements for structuring organ transplant programs. The discussion goes on to outline what a trustworthy program must be, and to provide a conceptual perspective for drawing lines and setting boundaries for ethically acceptable living donor transplantation. It also explains the morally relevant considerations for accepting emotionally related or unrelated living donors. PMID- 12764537 TI - Severe asthma: epidemiology, pathophysiology and treatment. AB - Severe asthma is poorly understood clinically, physiologically and pathologically. Whereas milder forms of asthma are generally easily treated, more severe forms often remain refractory to the best current medical care. Although some asthmatics have been severely affected for most of their lives, there appears to be a second group that develops severe disease in adulthood. Additionally, it is not clear which genetic and environmental elements may be most important in the development of severe disease. Physiologically, these patients often have airtrapping and may have loss of elastic recoil, as well. The pathology demonstrates a heterogeneity of findings, including continued eosinophilic inflammation, structural changes, and distal disease. Treatment is problematic and will probably remain so until a better understanding of this disease develops. PMID- 12764538 TI - Immunologic factors in transplant arteriopathy: insight from animal models. AB - Transplant arteriopathy is the leading cause of long-term morbidity and mortality following heart transplantation. The pathologic hallmark of this disease is intimal proliferation. Animal models have demonstrated that immunologic factors, including cytokines, cellular adhesion molecules and inflammatory cells, play a significant role in the development of this arteriopathy. One goal of future studies will be to translate findings in animal models into effective treatments in humans. PMID- 12764539 TI - Laboratory aspects of tick-borne diseases: lyme, human granulocytic ehrlichiosis and babesiosis. AB - Lyme disease, human granulocytic ehrlichiosis (HGE) and babesiosis are emerging infections in the northeastern and midwestern United States, where Ixodes scapularis ticks are prevalent. Lyme disease and babesiosis have also been reported on the West Coast, but less frequently. Lyme disease presents frequently with a skin lesion known as erythema migrans (EM), and diagnostic tests are not necessary if the lesion is classical. Those patients presenting without EM or with atypical skin lesions may need laboratory confirmation. The most frequently used laboratory modality consists of the 2-step serological assays, employing a sensitive ELISA as a first step, followed by IgG and/or IgM immunoblots. Current guidelines for interpretation are those recommended by the CDC. HGE and babesiosis are febrile illnesses with non-specific signs and symptoms. Both infections may present with routine laboratory abnormalities, including leukopenia and/or thrombocytopenia in HGE and anemia in babesiosis. Moderate elevations of liver enzymes may occur in all three tick-borne infections. Specific diagnostic modalities for acute-phase HGE include buffy coat smear examination, culture and PCR. Culture appears to have the greatest sensitivity of the three tests. Babesiosis can be diagnosed by peripheral blood examination for the intraerythrocytic parasites, PCR or serology. Co-infections with these agents exist, but they should be documented by detection of the organisms rather than by serology, since seroprevalence rates are high in endemic areas. PMID- 12764540 TI - Lyme disease trends--Dutchess County, New York, 1992-2000. AB - BACKGROUND: Lyme disease is a vector-borne infectious disease, accounting for more than 95% of all reported vector-borne illness in the United States. From 1992 2000, Dutchess County reported more cases of Lyme disease than any other county in the United States, consistently ranking among the top ten in incidence rates. We analyzed 1992-2000 Dutchess County Lyme disease surveillance data to characterize Lyme disease trends, identify high-risk populations, and examine the frequency of the characteristic lesion, erythema migrans. METHODS: A Lyme disease case was defined as a person with physician-diagnosed erythema migrans or at least one late manifestation of the disease, with laboratory confirmation. A surveillance database of cases reported in Dutchess County from 1992-2000 was obtained from the New York State Department of Health. Annual incidence rates by age, gender, race, ethnicity, and ZIP codes, and frequency of erythema migrans were calculated. RESULTS: From 1992 through 2000, a total of 9,548 cases of Lyme disease were reported by Dutchess County to the New York State Department of Health, for a crude mean annual incidence rate of 400 cases per 100,000 persons per year. The incidence rate peaked at 683/100,000 in 1996, and then declined from 1998 to 2000. A bimodal age distribution was seen, with the initial peak among children aged 5-9 years (617/100,000) and the second peak among adults aged 60-64 years (627/100,000). A male preponderance was clearly seen between the ages of 5-19 years, and beyond the age of 60 years. Highest incidence rates were reported in central Dutchess County. Onset of illness occurred most frequently in June, July, and August. Ninety-four percent of cases occurred among the predominantly white population, which had the highest incidence rate (431/100,000) among the races. Incidence rate for non-Hispanics was more than double that for Hispanics. Eighty-one percent of reported cases had erythema migrans. CONCLUSIONS: While some prevention programs could be broadly targeted to the entire Dutchess County population, other interventions might be most effective if they focused on the high-risk population groups and areas defined in this report. The high proportion of cases with erythema migrans suggests that early diagnosis and treatment should be effective in reducing late-stage complications of Lyme disease in Dutchess County. Surveillance data for other endemic counties and states can be similarly analyzed to enhance and monitor local prevention programs. PMID- 12764542 TI - Suitability of temperature-sensitive transponders to measure body temperature during animal experiments required for regulatory tests. AB - Body temperature is a clinical parameter in vaccine quality control to detect systemic side-effects or to monitor progression of infectious diseases. Moreover, changes in body temperature are used as clinical parameters to define humane endpoints in animal experiments. However, measuring body temperature via the rectal route can be troublesome and distressing to the animal. Non-invasive measurement methods were developed in recent years. The aim of this investigation was to study and to compare rectally measured body temperature with data obtained with implanted temperature-sensitive transponders (TST) in mice, guinea pigs, rabbits and pigs under the controlled conditions of regulatory testing. PMID- 12764541 TI - Comparison of the reactivity of human and rabbit blood towards pyrogenic stimuli. AB - A comparison between humans and rabbits was performed based on stimulation of whole blood with well-known pyrogens from Gram-negative and Gram-positive bacteria, such as lipopolysaccharide (LPS) and lipoteichoic acid (LTA), respectively. The reactivity was measured as release of IL-1 beta and IL-8 by ELISA. The reactivity of the two species towards LPS was found to be similar, whereas their reactivity towards LTA differed considerably. Differences between the levels of IL-1 beta and IL-8 release were observed in both species. This finding suggests that the In vitro Pyrogen Test (IPT) which uses human blood to detect contaminations, e.g. of injectable drugs, might predict the human reaction to the contamination better than the "gold standard" rabbit pyrogen test. PMID- 12764543 TI - Development of an in vitro model to study oxidative DNA damage in human coronary artery endothelial cells. AB - The endothelial cell layer is a multifunctional barrier between the blood stream and the vascular wall. Reactive oxygen species can damage endothelial cells and may so potentiate atherosclerotic lesion formation. Therefore, we established an in vitro model for the qualitative and quantitative investigation of oxidative DNA damage and repair in human coronary arterial endothelial cells (CAEC). Oxidative DNA damage was induced by standardised treatment with 50-400 microM hydrogen peroxide (H(2)O(2)). The amount of DNA damage was quantified by determination of DNA single strand breaks (SSB) and alkali-labile sites in individual cells, using the alkaline single-cell gel electrophoresis assay- "comet assay". Significant DNA damage could be induced reproducibly in CAEC cells after exposure to 50 microM H(2)O(2) for 10 min. Additional treatment with catalase prevented DNA damage by H(2)O(2). The time needed for DNA repair depended on the initial grade of damage. After 30 min post-incubation at 37 degrees C, DNA damage was completely repaired in cells treated with 50 microM H(2)O(2). Cell samples initially damaged with H(2)O(2) concentrations between 100 microM and 400 microM were repaired after 60 min. This endothelial cell culture model allows experiments on oxidative DNA damage, alteration, and repair directly on the relevant target cells. Animals are used neither as direct objects of such experiments nor as cell or tissue donors. PMID- 12764544 TI - Proof of principle: detection of genotoxicity by a fluorescence-based recombination test in mammalian cells. AB - Genotoxicity tests available today have several shortcomings. The widely applied Ames assay measures mutations in bacteria, thereby disregarding the physiological particularities of the human cell and organism.We provide first evidence for a new concept of genotoxicity detection in living human cell cultures. The data were obtained by use of a newly developed assay, which is based on the quantification of fluorescent signals, i.e. counting of the relative number of fluorescent cells in the sample. It is characterised by a short reaction time and fulfills the requirements for automated performance. The new system monitors chromosomal rearrangements and, therefore, is predicted to detect a broad spectrum of genotoxic substances. Indeed, we demonstrate the genotoxic effect of ionising radiation, of an Ames assay positive compound, and of two compounds which are poorly mutagenic in the Ames assay. The new assay will be optimised further and adapted to the requirements for routine analysis in order to help to further reduce animal experimentation in genotoxicity testing. PMID- 12764545 TI - Internet laboratory for predicting harmful effects triggered by drugs and chemicals--a progress report. AB - The main objective of our institution is to establish a virtual laboratory on the Internet to allow for a reliable in silico estimation of harmful effects triggered by drugs, chemicals and their metabolites. In the past two years, we have compiled a pilot system including the 3D models of five receptors known to mediate adverse effects (the Ah, 5HT(2A), cannabinoid, GABA(A), and estrogen receptor, respectively) and tested them against 280 compounds (drugs, chemicals, toxins). Within this set-up we could demonstrate that our concept is able to both recognise toxic compounds substantially different from those used in the training set as well as to classify harmless compounds clearly as being non-toxic at low level doses. This suggests that our approach can be used for the prediction of adverse effects of drug molecules and chemicals. It is the aim to provide free access to this 3D data base, particularly to universities, hospitals and regulatory bodies as it bears a significant potential to recognise hazardous compounds early in the development process and withdraw them from the evaluation pipeline. Hence, for substances recognised as hazardous in silico, subsequent toxicity tests involving animal models become obsolete. PMID- 12764546 TI - [Third statistical report on the numbers of laboratory animals used in the European Union--tendencies, problems, decisions]. AB - In 2003, the European Commission has published its third statistical report on the numbers of laboratory animals used in the European Union, in which the data of 1999 are analysed. For the first time, with the exception of Germany, the data presented were raised on the basis of a new registration scheme. The significance of these data is limited due to deficiencies of the tables used, restricted comparability with previously raised data, and lacking relevance to the current situation. Despite this imperfect basis, the European Commission concludes in its report that there has been a significant reduction in the number of laboratory animals used in the EU compared to the previous statistics. This statement must be regarded critically as several national statistics of the last years verify that the number of laboratory animals used has in fact increased. According to the comments of various national authorities included in the report, particularly genetic engineering is responsible for the increase in animal numbers. PMID- 12764548 TI - HbHope/HbS and HbS/beta-thal double compound heterozygosity in a Mauritanian family: clinical and biochemical studies. AB - The hemoglobin Hope was discovered in a Mauritanian family that comes from Gorgol in the southwest of the country. The family belongs to the Soninke ethnic group, which is one of the black population groups in Mauritania. Along with this abnormal hemoglobin, HbS and beta-thalassemia were also found. This family, which we refer to as D, was encountered during a survey we conducted to study hemoglobinopathies in Mauritania. First the father was identified to carry an association of HbS and HbHope, then the study was extended to the entire family members: the wife who was found to have a beta-thalassemia trait and their three children who were found to carry HbS/beta-thalassemia mutations each. All together this family carries three different mutations that resulted in a double compound heterozygosity HbHope/HbS and HbS/beta-thal. PMID- 12764547 TI - Spleen involvement in Hodgkin's lymphoma: assessment and risk profile. AB - Diagnostic laparotomy is no longer routinely performed in Hodgkin's lymphoma and noninvasive diagnosis of spleen involvement remains uncertain. In order to assess the probability of splenic involvement based on clinical parameters, we retrospectively analyzed data on patients of the German Hodgkin's Lymphoma Study Group (GHSG) who underwent staging laparotomy and for whom splenic weight and size were available. Our study included 376 patients with Hodgkin's lymphoma who underwent staging laparotomy and splenectomy according to the treatment policy of the GHSG between February 1981 and January 1993. Univariate and multivariate analyses of pretherapeutic clinical characteristics and splenic weight were performed in order to predict the probability of splenic involvement. Computed tomographic (CT) images of 25 patients were available and used to correlate radiological splenic size and pathological splenic weight. In 171 of 376 patients spleen involvement was found. Average weight of the spleens was 258 g (+/-257) ranging from 55 to 3290 g. All spleens with a weight above 2000 g showed disease involvement, while those under 150 g were never involved. In the multivariate analysis, splenic weight ( p<0.001), erythrocyte sedimentation rate ( p<0.001), and clinical stage ( p<0.01) were found to be independently prognostic for spleen involvement. Splenic weight was highly correlated with a spleen index defined as the product of length, width, and thickness measured by CT (correlation coefficient: 0.93). By applying the identified risk factors in clinically staged patients spleen involvement can be determined. Spleen weight can be estimated with the help of a spleen index. Above an index of 1000 the probability of spleen involvement is higher than 90%. This might be of outstanding importance for patients being scheduled for involved field radiation. PMID- 12764549 TI - Treatment and survival of 38 female breast lymphomas: a population-based study with clinical and pathological reviews. AB - Breast lymphomas are rare and consensus about their treatment is lacking. A population-based study of 38 breast lymphomas, registered in the databases of two Comprehensive Dutch Cancer Centers from 1981 to 1999, was performed. The median age of all female patients was 65 years (20-92): 25 patients had localized and 13 patients had disseminated lymphoma. The most common type was diffuse large B-cell lymphoma (DLBCL), which accounted for 17 of the localized and 4 of the disseminated cases. Burkitt's lymphoma (BL), three being disseminated, was found in four patients. There were six extranodal marginal zone lymphomas (ENMZL), three being localized. Seven DLBCL and one BL showed additional histological features of mucosa-associated lymphoid tissue (MALT) lymphoma. Localized aggressive lymphomas treated with surgery and/or radiation therapy had relapse rates of 100% and 67%, respectively. Cyclophosphamide, hydroxydaunomycin, vincristine, and prednisone (CHOP)-like chemotherapy with or without local irradiation led to 17% relapses in patients with localized aggressive lymphoma. Median follow-up time was 32 months (0.6-218); 37% of the patients relapsed and 24% had progressive disease. Response to salvage regimens, given to 91% of the patients with recurrent disease, was poor. The 2-year overall survival rate was 63%, 72% for patients with localized disease, and 46% for patients with disseminated lymphoma. The majority of breast lymphomas are localized aggressive lymphomas that should be treated initially with CHOP-like chemotherapy with or without irradiation. The initial choice of treatment is very important because response to salvage regimens is poor. PMID- 12764550 TI - Monoclonal and biclonal gammopathy in two patients infected with Bartonella henselae. AB - Two immunocompetent patients with cat-scratch disease due to infection with Bartonella henselae developed monoclonal and biclonal gammopathy. Neither patient had evidence of any other known cause of plasma cell dyscrasia, and antibiotic eradication of Bartonella henselae infection resulted in the prompt disappearance of the gammopathy. Hence, cat-scratch disease should be added to the list of possible underlying disorders in individuals presenting with monoclonal and biclonal gammopathy. PMID- 12764551 TI - Cerebral metabolic changes accompanying conversion of mild cognitive impairment into Alzheimer's disease: a PET follow-up study. AB - A high percentage of patients with mild cognitive impairment (MCI) develop clinical dementia of the Alzheimer type (AD) within 1 year. The aim of this longitudinal study was to identify characteristic patterns of cerebral metabolism at baseline in patients converting from MCI to AD, and to evaluate the changes in these patterns over time. Baseline and follow-up examinations after 1 year were performed in 22 MCI patients (12 males, 10 females, aged 69.8+/-5.8 years); these examinations included neuropsychological testing, structural cranial magnetic resonance imaging and fluorine-18 fluorodeoxyglucose positron emission tomography (PET) evaluation of relative cerebral glucose metabolic rate (rCMRglc). Individual PET scans were stereotactically normalised with NEUROSTAT software (Univ. of Michigan, Ann Arbor, USA). Subsequently, statistical comparison of PET data with an age-matched healthy control population and between patient subgroups was performed using SPM 99 (Wellcome Dept. of Neuroimaging Sciences, London, UK). After 1 year, eight patients (36%) had developed probable AD (referred to as MCI(AD)), whereas 12 (55%) were still classified as having stable MCI (referred to as MCI(MCI)). Compared with the healthy control group, a reduced rCMRglc in AD typical regions, including the temporoparietal and posterior cingulate cortex, was detected at baseline in patients with MCI(AD). Abnormalities in the posterior cingulate cortex reached significance even in comparison with the MCI(MCI) group. After 1 year, MCI(AD) patients demonstrated an additional bilateral reduction of rCMRglc in prefrontal areas, along with a further progression of the abnormalities in the parietal and posterior cingulate cortex. No such changes were observed in the MCI(MCI) group. In patients with MCI, characteristic cerebral metabolic differences can be delineated at the time of initial presentation, which helps to define prognostic subgroups. A newly emerging reduction of rCMRglc in prefrontal cortical areas is associated with the transition from MCI to AD. PMID- 12764553 TI - Antifungal mechanism of an anti-Pythium protein (SAP) from the marine bacterium Streptomyces sp. strain AP77 is specific for Pythium porphyrae, a causative agent of red rot disease in Porphyra spp. AB - Previously we reported an antifungal protein specific to Pythium porphyrae, a causative agent of red rot disease afflicting seaweed Porphyra spp. This study was carried out to identify the antifungal mechanism of the antifungal protein to P. porphyrae. When we first examined the effect of an anti- Pythium protein (SAP) on the P. porphyrae cell walls, SAP did not decompose the six structural polysaccharides in Pythium cell walls. However, hyphal growth was significantly inhibited in Pythium cells treated with 50 microg/ml of SAP by MTT assay. Protoplasmic leakage was observed in P. porphyrae hyphae treated with SAP for 1 h, followed by hyphal swelling and disintegration, using SYTOX Green, and SAP permeabilized the membrane of P. porphyrae in a dose-dependent manner. Treating P. porphyrae cells with SAP in the presence of carbonyl cyanide m chlorophenylhydrazone (CCCP), a membrane-depolarizing agent, significantly reduced the membrane permeability to SYTOX Green. Moreover, a similar effect was observed when the P. porphyrae cells were treated with SAP in the presence of MgCl2. In contrast, identical treatment in the presence of KCl significantly increased the membrane permeability to SYTOX Green. These results suggested that anti- Pythium mechanism of SAP was related to alteration of the membrane permeability in P. porphyrae. PMID- 12764554 TI - Catabolism of hydroxyacids and biotechnological production of lactones by Yarrowia lipolytica. AB - The gamma- and delta-lactones of less than 12 carbons constitute a group of compounds of great interest to the flavour industry. It is possible to produce some of these lactones through biotechnology. For instance, gamma-decalactone can be obtained by biotransformation of methyl ricinoleate. Among the organisms used for this bioproduction, Yarrowia lipolytica is a yeast of choice. It is well adapted to growth on hydrophobic substrates, thanks to its efficient and numerous lipases, cytochrome P450, acyl-CoA oxidases and its ability to produce biosurfactants. Furthermore, genetic tools have been developed for its study. This review deals with the production of lactones by Y. lipolytica with special emphasis on the biotransformation of methyl ricinoleate to gamma-decalactone. When appropriate, information from the lipid metabolism of other yeast species is presented. PMID- 12764555 TI - Phytoremediation: an overview of metallic ion decontamination from soil. AB - In recent years, phytoremediation has emerged as a promising ecoremediation technology, particularly for soil and water cleanup of large volumes of contaminated sites. The exploitation of plants to remediate soils contaminated with trace elements could provide a cheap and sustainable technology for bioremediation. Many modern tools and analytical devices have provided insight into the selection and optimization of the remediation process by plant species. This review describes certain factors for the phytoremediation of metal ion decontamination and various aspects of plant metabolism during metallic decontamination. Metal-hyperaccumulating plants, desirable for heavily polluted environments, can be developed by the introduction of novel traits into high biomass plants in a transgenic approach, which is a promising strategy for the development of effective phytoremediation technology. The genetic manipulation of a phytoremediator plant needs a number of optimization processes, including mobilization of trace elements/metal ions, their uptake into the root, stem and other viable parts of the plant and their detoxification and allocation within the plant. This upcoming science is expanding as technology continues to offer new, low-cost remediation options. PMID- 12764552 TI - PET-based molecular imaging in neuroscience. AB - Positron emission tomography (PET) allows non-invasive assessment of physiological, metabolic and molecular processes in humans and animals in vivo. Advances in detector technology have led to a considerable improvement in the spatial resolution of PET (1-2 mm), enabling for the first time investigations in small experimental animals such as mice. With the developments in radiochemistry and tracer technology, a variety of endogenously expressed and exogenously introduced genes can be analysed by PET. This opens up the exciting and rapidly evolving field of molecular imaging, aiming at the non-invasive localisation of a biological process of interest in normal and diseased cells in animal models and humans in vivo. The main and most intriguing advantage of molecular imaging is the kinetic analysis of a given molecular event in the same experimental subject over time. This will allow non-invasive characterisation and "phenotyping" of animal models of human disease at various disease stages, under certain pathophysiological stimuli and after therapeutic intervention. The potential broad applications of imaging molecular events in vivo lie in the study of cell biology, biochemistry, gene/protein function and regulation, signal transduction, transcriptional regulation and characterisation of transgenic animals. Most importantly, molecular imaging will have great implications for the identification of potential molecular therapeutic targets, in the development of new treatment strategies, and in their successful implementation into clinical application. Here, the potential impact of molecular imaging by PET in applications in neuroscience research with a special focus on neurodegeneration and neuro-oncology is reviewed. PMID- 12764556 TI - Entomopathogenic fungi as biological control agents. PMID- 12764557 TI - A novel process for the production of a veterinary rabies vaccine in BHK-21 cells grown on microcarriers in a 20-l bioreactor. AB - We studied BHK-21 cells growth in a 2-l bioreactor and investigated the effects of microcarrier concentration, type of growth medium, culture mode and serum concentration. The highest cell density reached was equal to 4x10(6) cells/ml and was achieved in minimum essential medium supplemented with Hanks' salts, non essential amino acids and 5% fetal calf serum, using a perfusion culture mode and a microcarrier concentration of 4 g Cytodex 3/l. We studied rabies virus production (PV/BHK-21 strain) by BHK-21 cells grown at the optimal conditions determined previously. We analyzed the effects of multiplicity of infection (MOI) and type of medium used for virus multiplication in spinner-flasks and showed that the highest virus titer reached (when the cells were infected at a MOI of 0.3) in M199 medium supplemented with 0.2% of bovine serum albumin was equal to 8.2x10(7) Fluorescent Focus Units (FFU)/ml. When we grew the cells in a 2-l perfused bioreactor, we obtained a maximal virus titer of 3x10(8) FFU/ml. In addition, we scaled-up to a 20-l bioreactor and obtained similar results for cell density and virus titer. The experimental vaccine we developed meets WHO requirements for vaccine potency. Each run yielded about 40,000 doses of potent vaccine. PMID- 12764558 TI - Effect of electrical stimulation on HIV-1-infected HeLa cells cultured on an electrode surface. AB - Acquired immunodeficiency syndrome (AIDS) is a disease caused by infection with the human immunodeficiency virus (HIV). Although drug therapy for AIDS is available, problems such as side effects associated with drug therapy and the appearance of resistant HIV strains have arisen. Therefore, therapies based on new principles other than drug treatment are required. In the present study, the effect of electrical stimulation on HIV-1(LAI) chronically infected HeLa (P6 HeLa/HIV-1(LAI)) cells cultured on an electrode surface was examined. The results indicated that sensitivity to electrical stimulation was much higher in P6 HeLa/HIV-1(LAI) cells than in uninfected P6 HeLa cells. When electrical stimulation was applied at 1.0 V (vs. Ag/AgCl) for 20 min, the proportion of damage to cell membrane among P6 HeLa/HIV-1(LAI) cells, as evaluated by Trypan blue staining, was approximately 4 times higher than that for uninfected P6 HeLa cells. Furthermore, in comparison with uninfected P6 HeLa cells, the proliferation of P6 HeLa/HIV-1(LAI) cells was significantly suppressed after electrical stimulation. This technique was proven to selectively kill P6 HeLa/HIV 1(LAI) cells, when compared with uninfected control cells. PMID- 12764559 TI - Nutrient regulation of epothilone biosynthesis in heterologous and native production strains. AB - Fermentation media with different initial concentrations of ammonium and phosphate salts were used to study the inhibitory effects of those ions on growth and production of epothilone in Sorangium cellulosum and Myxococcus xanthus. The native epothilone producer, S. cellulosum was more sensitive to ammonium and phosphate than the heterologous producer, M. xanthus. An ammonium concentration of 12 mM reduced epothilone titers by 90% in S. cellulosum but by only 40% in M. xanthus. When 5 mM phosphate was added to the medium, production in both strains was 60% lower. Higher phosphate concentrations had little additional effect on M. xanthus titers, but epothilone production with 17 mM extra-cellular phosphate in S. cellulosum was 95% lower than in the control condition. The effect of iron supplementation to the fermentation medium was also investigated. Both strains showed best production with 20 microM iron added to the medium. PMID- 12764560 TI - Optimization of the extracellular production of a bacterial phytase with Escherichia coli by using different fed-batch fermentation strategies. AB - The extracellular production of Escherichia coli phytase was studied in fed-batch fermentations. Two different feeding strategies were compared: control by keeping the glucose concentration constant, and control by keeping a low constant oxygen level in the medium. For the feeding control based on glucose concentration, a recently developed rapid glucose controlling system was tested for the first time in bacterial cultivations and used to establish the fermentative production of extracellular phytase with E. coli. High activity levels (120 U ml(-1)) at short cultivation times (14 h) were obtained. Even higher activity levels - albeit at longer cultivation times - were reached by applying a feeding control, the main characteristic of which was a constant low oxygen concentration. The optimum oxygen level for the production of phytase was in the range of 5-10% saturation. PMID- 12764561 TI - Biochemical characterization and antifungal activity of an endo-1,3-beta glucanase of Paenibacillus sp. isolated from garden soil. AB - A 44-kDa 1,3-beta-glucanase was purified from the culture medium of a Paenibacillus strain with a 28-fold increase in specific activity with 31% recovery. The purified enzyme preferentially catalyzes the hydrolysis of glucans with 1,3-beta-linkage and has an endolytic mode of action. The enzyme also showed binding activity to various insoluble polysaccharides including unhydrolyzable substrates such as xylan and cellulose. The antifungal activity of this Paenibacillus enzyme and a previously purified 1,3-beta-glucanase from Streptomyces sioyaensis were examined in this study. Both enzymes had the ability to damage the cell-wall structures of the growing mycelia of phytopathogenic fungi Pythium aphanidermatum and Rhizoctonic solani AG-4. Nonetheless, the Paenibacillus enzyme had a much stronger effect on inhibiting the growth of fungi tested. PMID- 12764562 TI - Esterase EstE from Xanthomonas vesicatoria ( Xv_EstE) is an outer membrane protein capable of hydrolyzing long-chain polar esters. AB - A new esterase gene from Xanthomonas vesicatoria (formerly X. campestris) DSM 50861 was identified, cloned from a chromosomal gene library and overexpressed in Escherichia coli. The corresponding DNA fragment contains an ORF of 1,818 bp, encoding a hydrolase of the GDSL esterase family. A protein of about 67 kDa, named Xv_EstE, was expressed from this fragment. A N-terminal signal peptide was processed under low-expression conditions, yielding a 63-kDa mature protein. The predicted amino acid sequence showed distinct homology to esterases of the GDSL family. Based on homology, a catalytic triad Gly-Asp-Ser could be defined. Amino acid sequence alignments and computer-assisted structure prediction indicated the presence of a carboxyl-terminal beta-barrel membrane domain which might facilitate binding of Xv_EstE to the outer membrane. This could be verified by differential cell fractionation experiments, in which Xv_EstE was exclusively found in the outer membrane fraction. Xv_EstE showed preferential hydrolytic activity on short chain (up to C(8)) and para-substituted nitrophenylesters as substrates. However, only long-chain 1-hydroxy-pyrene-3,6,8-trisulfonic acid (HPTS)-fatty acid esters were hydrolyzed. Xv_EstE was also found to be active on a series of substrates of industrial interest, such as 1-methylprop-2-ynyl acetate, for which an enantioselectivity up to 93% ee could be recognized. PMID- 12764563 TI - Structure-function study of the amino-terminal stretch of the catalase subunit molecule in oligomerization, heme binding, and activity expression. AB - Analysis of the protein structure of bovine liver catalase suggested that the N terminal region containing two alpha-helices may function as a linker binding to another subunit. The number of amino-acid residues in catalase from the n-alkane assimilating yeast Candida tropicalis (CTC) is the lowest of any eukaryotic catalase molecule hitherto investigated, and only one helix, corresponding to the helix alpha2 in bovine liver catalase, is estimated to be present in the same region. In the present study, N-terminal-deleted mutants of CTC were characterized to evaluate the role of the alpha-helix structure in the N-terminal region. CTCDelta1-4 and CTCDelta1-24, whose N-terminal regions were shortened by four and 24 amino-acid residues, respectively, showed an 80% decrease in specific activity compared to wild-type CTC in spite of containing the same amount of heme as in the wild-type. Polyacrylamide gel electrophoresis under nondenaturing conditions revealed that the mutants contained large amounts of oligomeric forms with molecular masses less than 220 kDa (tetramer assembly). Although the smaller oligomers were found to be bound with heme, only the tetramer exhibited catalase activity in activity staining on nondenaturing gel. CTCDelta1-49, a mutant with deletion of the N-terminal 49 amino-acid residues which contain the conserved helix alpha2, showed no catalase activity and no heme binding. However, the CD spectrum profiles of CTCDelta1-49, CTCDelta1-4, and CTCDelta1-24 indicated that these mutant subunits could attain secondary conformations similar to that of wild-type CTC, regardless of their binding with heme. From these results, it was concluded that the N-terminal stretch of catalase is significant for complete assembly into active tetramer and that the conserved helix alpha2, although it has little effect on the formation of the subunit secondary structure, is indispensable not only in assembling tetramer but also in binding heme. PMID- 12764564 TI - Medium and copy number effects on the secretion of human proinsulin in Escherichia coli using the universal stress promoters uspA and uspB. AB - The use of the uspA and uspB promoters (universal stress promoters) for heterologous protein production in Escherichia coli is described. Best results were obtained with a moderate copy number vector (15-60 copies) bearing the uspA promoter, reaching 4.6 mg/g dry cell weight (DCW) of ZZ-proinsulin secreted to the periplasm and 1.9 mg/g DCW secreted to the culture medium. These values are about 1.7-fold higher than those previously reported with the same ZZ fusion tag and the SpA leader peptide showing that these stress promoters are potentially valuable for recombinant protein secretion in E. coli. It is further demonstrated that the use of M9 minimal medium is advantageous for protein secretion as compared to LB rich medium. PMID- 12764565 TI - Expression of the Aspergillus niger glucose oxidase gene in Saccharomyces cerevisiae and its potential applications in wine production. AB - There is a growing consumer demand for wines containing lower levels of alcohol and chemical preservatives. The objectives of this study were to express the Aspergillus niger gene encoding a glucose oxidase (GOX; beta- d-glucose:oxygen oxidoreductase, EC 1.1.3.4) in Saccharomyces cerevisiae and to evaluate the transformants for lower alcohol production and inhibition of wine spoilage organisms, such as acetic acid bacteria and lactic acid bacteria, during fermentation. The A. niger structural glucose oxidase (gox) gene was cloned into an integration vector (YIp5) containing the yeast mating pheromone alpha-factor secretion signal (MFalpha1(S)) and the phosphoglycerate-kinase-1 gene promoter (PGK1(P)) and terminator (PGK1(T)). The PGK1(P)- MFalpha1(S)- gox- PGK1(T) cassette (designated GOX1) was introduced into a laboratory strain (Sigma1278) of S. cerevisiae. Yeast transformants were analysed for the production of biologically active glucose oxidase on selective agar plates and in liquid assays. The results indicated that the recombinant glucose oxidase was active and was produced beginning early in the exponential growth phase, leading to a stable level in the stationary phase. The yeast transformants also displayed antimicrobial activity in a plate assay against lactic acid bacteria and acetic acid bacteria. This might be explained by the fact that a final product of the GOX enzymatic reaction is hydrogen peroxide, a known antimicrobial agent. Microvinification with the laboratory yeast transformants resulted in wines containing 1.8-2.0% less alcohol. This was probably due to the production of d glucono-delta-lactone and gluconic acid from glucose by GOX. These results pave the way for the development of wine yeast starter culture strains for the production of wine with reduced levels of chemical preservatives and alcohol. PMID- 12764566 TI - Deletion of scbA enhances antibiotic production in Streptomyces lividans. AB - Antibiotic production in many streptomycetes is influenced by extracellular gamma butyrolactone signalling molecules. In this study, the gene scbA, which had been shown previously to be involved in the synthesis of the gamma-butyrolactone SCB1 in Streptomyces coelicolor A3(2), was deleted from the chromosome of Streptomyces lividans 66. Deletion of scbA eliminated the production of the antibiotic stimulatory activity previously associated with SCB1 in S. coelicolor. When the S. lividans scbA mutant was transformed with a multi-copy plasmid carrying the gene encoding the pathway-specific activator for either actinorhodin or undecylprodigiosin biosynthesis, production of the corresponding antibiotic was elevated significantly compared to the corresponding scbA(+) strain carrying the same plasmid. Consequently, deletion of scbA may be useful in combination with other strategies to construct host strains capable of improved bioactive metabolite production. PMID- 12764567 TI - Cloning and sequence analysis of the ces10 gene encoding a Sphingomonas paucimobilis esterase. AB - The ces10 gene of the gellan gum-producing strain Sphingomonas paucimobilis ATCC 31461 was cloned and sequenced. Multi-sequence alignment of the deduced protein indicated that Ces10 belongs to the serine hydrolase family with a potential catalytic triad comprising Ser(153) (within the G-X-S-X-G consensus sequence), His(75) and Asp(125). The mixed block results obtained following pattern search and the low identities detected in a BLAST analysis indicate that Ces10 is significantly different from other characterised bacterial esterases/lipases. Nevertheless, the Ces10 amino acid sequence showed 45% similarity with Rhodococcus sp. heroin esterase and 48% with Bacillus subtilis p-nitrobenzyl esterase. Ces10, with a predicted molecular mass of 30,641 Da, was overproduced in Escherichia coli and purified to homogeneity in a histidine-tagged form. Enzyme assays using p-nitrophenyl-esters (p-NP-esters) with different acyl chain lengths as the substrate confirmed the anticipated esterase activity. Ces10 exhibited a marked preference for short-chain fatty acids, yielding the highest activity with p-NP-propionate (optimal pH 7.4, optimal temperature 37 degrees C). PMID- 12764568 TI - Host-vector system for phenol-degrading Rhodococcus erythropolis based on Corynebacterium plasmids. AB - The strain Rhodococcus erythropolis CCM2595, which was shown to degrade phenol, was chosen for genetic studies. To facilitate strain improvement using the methods of gene manipulation, the technique of genetic transfer was introduced and cloning vectors were constructed. Using the plasmid pFAJ2574, an electrotransformation procedure yielding up to 7x10(4) transformants/microg DNA was optimized. Escherichia coli- R. erythropolis shuttle vectors were constructed using the replicons pSR1 and pGA1 from Corynebacterium glutamicum. The small vector pSRK21 (5.8 kb) provides six unique cloning sites and selection of recombinant clones using alpha-complementation of beta-galactosidase in E. coli. This vector, exhibiting high segregational stability under non-selective conditions in R. erythropolis CCM2595, was applied to cloning and efficient expression of the gene coding for green fluorescent protein (gfpuv). PMID- 12764569 TI - Functional analyses of genes involved in the metabolism of ferulic acid in Pseudomonas putida KT2440. AB - Pseudomonas putida KT2440 is a physiologically extremely versatile non-pathogenic bacterium that is applied as a "biosafety strain" in biotechnological processes, as authorized by the USA National Institute of Health. Analysis of the P. putida KT2440 whole-genome sequence revealed the genetic organization of the genes fcs, ech, and vdh, which are essential for ferulic acid conversion to vanillic acid via vanillin. To confirm the physiological function of these structural genes as feruloyl-CoA synthetase (Fcs), enoyl-CoA hydratase/aldolase (Ech), and vanillin dehydrogenase (Vdh), respectively, they were cloned and expressed in Escherichia coli. Recombinant strains harboring fcs and ech were able to transform ferulic acid to vanillin. The enzyme activities of Fcs and Vdh were determined in protein extracts of these cells. The essential involvement of fcs, ech and vdh in the catabolism of ferulic acid in P. putida KT2440 was proven by separately inactivating each gene by insertion of Omega-elements. The corresponding mutant strains KT2440 fcsOmegaKm, KT2440 echOmegaKm, and KT2440 vdhOmegaKm were not able to grow on ferulic acid. The potential application of P. putida KT2440 and the mutant strains in biotechnological vanillin production process is discussed. PMID- 12764570 TI - Growth and magnetosome formation by microaerophilic Magnetospirillum strains in an oxygen-controlled fermentor. AB - Media and growth conditions were optimized for the microaerobic cultivation of Magnetospirillum gryphiswaldense in flasks and in a fermentor, resulting in significantly increased cell and magnetosome yields, compared with earlier studies. A reliable method was established for the automatic control of low dissolved oxygen tensions (pO(2)) in the fermentor (oxystat). Growth and magnetosome formation by M. gryphiswaldense, M. magnetotacticum and Magnetospirillum sp. AMB-1 were studied at various oxygen concentrations. Despite differences in their growth responses with respect to oxygen, we found a clear correlation between pO(2) and magnetosome formation in all three Magnetospirillum strains. Magnetite biomineralization was induced only below a threshold value of 20 mbar O(2) and optimum conditions for magnetosome formation were found at a pO(2) of 0.25 mbar (1 bar = 10(5) Pa). A maximum yield of 6.3 mg magnetite l(-1) day(-1) was obtained with M. gryphiswaldense grown under oxystat conditions, which is the highest magnetosome productivity reported so far for a magnetotactic bacterium. In conclusion, the presented results provide the basis for large-scale cultivation of magnetospirilla under defined conditions. PMID- 12764571 TI - Interactions between irradiance and nutrient availability during astaxanthin accumulation and degradation in Haematococcus pluvialis. AB - Fully synchronised germination of Haematococcus pluvialis astaxanthin-replete aplanospores was induced by transfer to nitrogen-sufficient conditions under either high or low light intensities, and growth, pigment content and nitrogen consumption were monitored during the cell cycle. No germination of the aplanospores was achieved in the absence of nitrate, even when cells were transferred at low light intensities. On the other hand, cell density and chlorophyll concentration increased dramatically and astaxanthin concentration decreased in N-sufficient cultures due to the germination of 100% of the aplanospores, as demonstrated by flow cytometry. No significant effect of light intensity was observed on the degradation of astaxanthin during germination. In germinated cultures, nitrogen was depleted more rapidly under high light conditions, which resulted in earlier entry into the aplanospore stage and accumulation of astaxanthin. Germination of aplanospores accompanied by astaxanthin degradation could also be obtained in the dark in nutrient-sufficient conditions although at a much lower efficiency. The results demonstrate that nutrient availability is the main factor controlling the transition between red and green stages of H. pluvialis, with astaxanthin being accumulated only when cell division has ceased. High light levels accelerate the process by increasing the rate of nutrient depletion and providing more energy for astaxanthin synthesis. PMID- 12764572 TI - The ability of Bipolaris sorokiniana to modify geraniol and (-)-alpha-bisabolol as exogenous substrates. AB - The biocatalytic potential of Bipolaris sorokiniana was investigated in its ability to modify the monoterpene geraniol and the sesquiterpene alpha-bisabolol as exogenous substrates, using phosphate buffer as reaction medium. The cultures showed a promising oxidative profile, with conversion of geraniol to 6-methyl-5 hepten-2-one (74.9% yield) in a 5-day incubation and alpha-bisabolol to bisabolol oxide B (84.2% yield), in a 7-day incubation. PMID- 12764573 TI - Elemental compositions and characteristics of aerobic granules cultivated at different substrate N/C ratios. AB - The effects of the substrate N/C ratios on the formation, elemental compositions and characteristics of aerobic granules were investigated in four sequencing batch reactors. Results showed that aerobic granules could form at substrate N/C ratios ranging from 5/100 to 30/100 and the substrate N/C ratio had a direct and profound effect on the elemental compositions and characteristics of the aerobic granules. Nitrifying populations in aerobic granules were enriched significantly with the increase in the substrate N/C ratio, while the respective ratio of cell oxygen, nitrogen and calcium to cell carbon were also determined by the substrate N/C ratio. It was found that cell hydrophobicity of aerobic granules was inversely related to the ratio of cell oxygen normalized to cell carbon. Since the cell calcium content in aerobic granules developed at different substrate N/C ratios was even lower than that in the seed sludge, it is reasonable to conclude that the cell calcium would not contribute to aerobic granulation. This study probably for the first time demonstrates that the elemental composition, microbial distribution and characteristics of aerobic granules are related to the substrate N/C ratio applied. PMID- 12764574 TI - Predicting the effects of chlorine on the micro-organisms of filamentous bulking activated sludges. AB - Rapid and definite assessment of the effect that a specific biocide has on a specific case of filamentous bulking sludge is a much-needed tool in activated sludge wastewater treatment. The Live/Dead stain (LIVE/DEAD BacLight) distinguishing "living" and "non-living" cells, a nitrifying activity (NA) test and the oxygen uptake rate (OUR) measurement were examined for their appropriateness to predict the effects of chlorine on filamentous bulking sludges. The study showed the live/dead stain to be relevant for revealing the specific effect of chlorine on the filamentous bacteria of a bulking sludge. However, using live/dead stain alone for the determination of the appropriate chlorine dose against bulking may lead to an underestimation of the damage caused by chlorine to the useful microorganisms in the flocs. Indeed, using the live/dead stain, it was not easy to distinguish dead cells caused by chlorination from those originally present in the flocs The NA test was the most sensitive in detecting damage caused by chlorine to the floc-forming microorganisms. Therefore, for a safer determination of the chlorine dose effective against bulking and protective of the microbial activity of the sludge, the results of this study suggest coupling of the live/dead stain with the NA test and/or the OUR test. PMID- 12764575 TI - Adolescent-onset nicotine self-administration modeled in female rats. AB - RATIONALE: Although the great majority of tobacco addiction begins during adolescence, little is known about differential nicotine effects in adolescents versus adults. OBJECTIVES: A rat model was used to determine the impact of the age of onset on nicotine self-administration. METHODS: In expt 1, nicotine self administration of female Sprague-Dawley rats over a range of acute doses (0.01 0.08 mg/kg per infusion) was determined in adolescent (beginning at 54-62 days) versus adult (beginning at 84-90 days). In expt 2, chronic nicotine self administration over 4 weeks from adolescence into adulthood was compared with the chronic self-administration beginning in adulthood. In expt 3, adolescent-adult differences in nicotine effects on body temperature and locomotor responses were determined. RESULTS: Adolescent-onset rats showed a significant main effect of increased nicotine intake compared with adult-onset rats in an eight-fold range of acute unit doses/infusion. Significant age differences were also seen in the chronic level of nicotine self-administration. Over 4 weeks, the adolescent-onset group had nearly double the rate of nicotine self-administration of the benchmark nicotine dose (0.03 mg/kg per infusion) compared to the adult-onset group. This increased nicotine intake persisted into adulthood. Adolescent rats had significantly greater response than adults to the hypothermic effects of nicotine, but had significantly less response than adults to the reduction in locomotor activity seen after nicotine. CONCLUSIONS: Adolescent-onset nicotine self-administration in female rats was associated with significantly higher levels of nicotine self-administration versus rats, which began nicotine self administration in adulthood. This greater self-administration persists into adulthood and may underlie greater propensity of adolescents to nicotine addiction. PMID- 12764576 TI - New selective ligands of human cloned melatonin MT1 and MT2 receptors. AB - Melatonin has a key role in the circadian rhythm relay to periphery organs. Melatonin exerts its multiple roles mainly through two seven transmembrane domain, G-coupled receptors, namely MT1 or MT2 receptors. A pharmacological characterization of these human cloned melatonin hMT1 and hMT2 receptors stably expressed in HEK-293 or CHO cells is presented using a 2-[125I]-iodo-melatonin binding assay and a [35S]-GTPgammaS functional assay. Both reference compounds and new chemically diverse ligands were evaluated. Binding affinities at each receptor were found to be comparable on either HEK-293 or CHO cell membranes. Novel non-selective or selective hMT1 and hMT2 ligands are described. The [35S] GTPgammaS functional assay was used to define the functional activity of these compounds which included partial, full agonist and/or antagonist activity. None of the compounds acted as an inverse agonist. We report new types of selective antagonists, such as S 25567 and S 26131 for MT1 and S 24601 for MT2. These studies brought other new molecular tools such as the selective MT1 agonist, S 24268, as well as the non-selective antagonist, S 22153. Finally, we also discovered S 25150, the most potent melatonin receptor agonist, so far reported in the literature. PMID- 12764577 TI - Variations in process and outcomes of diabetes care by socio-economic status in Salford, UK. AB - AIMS/HYPOTHESIS: Our aim was to investigate trends in provision and outcomes of care by socio-economic status among patients with diabetes in Salford, United Kingdom. METHODS: Salford is a deprived urban area in North West England. Data for people with diabetes who were younger than 20 years of age (4034 patients in the year 1993 and 5671 by the year 2000) were extracted from the Diabetes Information System. Age-standardised means, proportions and attainment of targets were calculated for: processes of care indicators, intermediate outcomes and prescribing of preventive drug treatments in 1993 to 1994 and in 2000 to 2001 by quintiles of Townsend deprivation score. We did comparisons of means and proportions using age-adjusted linear regression and of trends using generalised estimating equations. Rate ratios for first microvascular and first macrovascular complication were estimated from proportional hazards models. RESULTS: Marked improvements occurred in all indicators. For patients managed in primary care, blood pressure and cholesterol measurement increased from 53% to 64% (p<0.001) and 27% to 61% (p<0.001) respectively; whilst mean systolic blood pressure decreased from 147 to 140 mmHg (p<0.001) and cholesterol concentrations from 6.0 to 5.1 mmol/l (p<0.001). Mean HbA(1c) increased from 7.8 to 8.1% (p<0.001). Prescribing of aspirin, anti-hypertensive and lipid-lowering drugs increased greatly. Trends varied little by socio-economic status. Patients from more affluent areas generally received more frequent clinical monitoring and preventive treatments, and had a lower BMI (29.5 vs 30.2 kg/m(2); p=0.009) and HbA(1c) (7.8 vs 8.2% p=0.006), though risks of first microvascular or macrovascular complications were similar. CONCLUSION/INTERPRETATION: Improvements in process and outcomes of care are possible for patients from all socio-economic groups. Socio-economic deprivation does not preclude high quality diabetes care. PMID- 12764578 TI - Gastric inhibitory polypeptide (GIP) dose-dependently stimulates glucagon secretion in healthy human subjects at euglycaemia. AB - AIMS/HYPOTHESIS: In the isolated perfused pancreas, gastric inhibitory polypeptide (GIP) has been shown to enhance glucagon secretion at basal glucose concentrations, but in healthy humans no glucagonotropic effect of GIP has yet been reported. Therefore, we studied the effect of GIP on glucagon secretion under normoglycaemic conditions. METHODS: Ten healthy subjects (9 men, 1 woman; age 33+/-11; BMI 26.8+/-2.2 kg/m(2)) received three different doses of intravenous GIP (7, 20, and 60 pmol/kg body weight) and placebo. Venous blood samples were drawn over 30 min for glucagon and GIP concentrations (specific radioimmunoassays). In addition, 31 healthy subjects (16 men, 15 women; 42+/-11 years; BMI 24.4+/-2.7 kg/m(2)) were studied with 20 pmol GIP/kg. Statistics were done with RM-ANOVA and Duncan's post hoc tests. RESULTS: Gastric inhibitory polypeptide dose-dependently stimulated glucagon secretion ( p=0.019) with a maximal increment after 10 min. Incremental glucagon concentrations (Delta(10-0 min)) were 0.1+/-0.7, 1.4+/-0.5, 2.4+/-0.5, and 3.4+/-0.8 pmol/l (for placebo and for 7, 20, and 60 pmol GIP/kg, respectively; p=0.017). After the injection of 20 pmol GIP/kg b.w. in 31 healthy subjects, glucagon concentrations increased over the baseline from 7.5+/-0.5 to 9.3+/-0.7 pmol/l ( p=0.0082). CONCLUSIONS/INTERPRETATION: Glucagon secretion is dose-dependently stimulated by GIP at basal glucose concentrations. The absence of a glucagonotropic GIP effect in previous studies could be due to the hyperglycaemic conditions used in these experiments. Our results underline differences between GIP and the glucagonostatic incretin GLP-1. PMID- 12764579 TI - Promotion of corneal epithelial wound healing in diabetic rats by the combination of a substance P-derived peptide (FGLM-NH2) and insulin-like growth factor-1. AB - AIMS/HYPOTHESIS: The healing of corneal epithelial wounds is often delayed in individuals with diabetes. The effect of the combination of a substance P-derived tetrapeptide (phenylalanine-glycine-leucine-methionine amide, or FGLM-NH(2)) and insulin-like growth factor-1 (IGF-1) on corneal epithelial wound healing was investigated in rats with streptozotocin-induced diabetes. METHODS: The corneal epithelium of diabetic and non-diabetic rats was removed, and the animals were treated by the application of eye drops containing FGLM-NH(2) and IGF-1, or vehicle alone as a control, six times a day for 3 days. The area of the corneal epithelial wound was measured at various times up to 72 h after removal of the corneal epithelium. RESULTS: The rate of corneal epithelial wound healing was slower in diabetic rats treated with vehicle than in non-diabetic rats. However, the rate of wound closure in diabetic rats treated with FGLM-NH(2) and IGF-1 was markedly increased compared with that in diabetic rats treated with vehicle. The wound healing process seemed similar in normal rats and in diabetic rats treated with FGLM-NH(2) and IGF-1. CONCLUSION/INTERPRETATION: The combination of FGLM NH(2) and IGF-1 promotes corneal epithelial wound healing in diabetic rats, suggesting that such a treatment might prove effective in humans with diabetic keratopathy. PMID- 12764581 TI - Alien genes introgression and development of monosomic alien addition lines from Oryza latifolia Desv. to rice, Oryza sativa L. AB - Oryza latifolia, a tetraploid wild relative of cultivated rice is an important source of resistance to bacterial blight (BB), the brown planthopper (BPH) and the whitebacked planthopper (WBPH). Interspecific hybrids were obtained between an elite breeding line (IR31917-45-3-2) of Oryza sativa (2n=24 AA) and O. latifolia Acc. No. 100914 (2n=48 CCDD). The crossability in F1 was 7.58% and it ranged from 0.11 to 0.62 in backcross generations. The F1 hybrid showed 2-6 II, 0 2 III, 0-1 IV and 22-32 I; the mean being 3.92 II + 0.11 III + 0.02 IV + 27.30 I per cell at diakinesis. Monosomic alien addition lines (MAALs) having a 2n chromosome complement of O. sativa and one chromosome of O. latifolia were characterized based on morphology and isozyme banding pattern. The MAALs were designated as MAAL-1, MAAL-2, MAAL-4, MAAL-5, MAAL-6, MAAL-7, MAAL-8, MAAL-9, MAAL-10, MAAL-11 and MAAL-12. The female transmission rates of the alien chromosome varied from 4.4 to 35.5%, whereas 8 of the 11 MAALs transmitted the alien chromosome through the male gamete, the range being 1.7% (MAAL 10) to 11.9% (MAAL 12). Disomic progenies in BC3 and BC4 generations had complete resemblance to the O. sativa parent. Of the 2,295 disomic BC3F3 progenies, 309 showed introgression for resistance to BPH and 188 each for WBPH and BB resistance. Four plant progenies which were resistant to both BPH and WBPH were also resistant to BB race 2 of the Philippines. Nine of the 34 BC3F1 plants showed introgression for ten allozymes of O. latifolia, such as Est5, Amp1, Pgi1, Mdh3, Pgi2, Amp3, Pgd2, Est9, Amp2 and Sdh1, located on 8 of the 12 chromosomes. Alien introgression was also detected for morphological traits such as long awns, earliness, black hull, purple stigma and apiculus. Abnormal plants with many wild species traits suddenly appeared in normal disomic progenies. These plants showing instability and abnormal segregation behaviour are being investigated for the activation of transposons. PMID- 12764582 TI - [Flat-panel detectors in X-ray systems]. AB - For all application segments X-ray systems with flat-panel detectors increasingly enter the market. In digital radiography,mammography and cardiologic angiography flat-panel detectors are already well established while they are made ready for market introduction in general angiography and fluoroscopy. Two flat-panel detector technologies are available. One technology is based on an indirect conversion process of X-rays while the other one uses a direct conversion method. For radiography and dynamic applications the indirect method provides substantial advantages, while the direct method has some benefits for mammography. In radiography and mammography flat-panel detectors lead to clear improvements with respect to workflow, image quality and dose reduction potentials. These improvements are fostered by the immediate availability of the image, the large dynamic range and the high sensitivity to X-rays. New applications and the use of complex image processing algorithms have the potential to enlarge the present diagnostic range of applications. Up to now, image intensifiers are still the well-established technology for angiography and fluoroscopy. Nevertheless flat panel detectors begin to enter this field, especially in cardiologic angiography. Characteristics of flat-panel detectors such as the availability of distortion free images, the excellent contrast resolution, the large dynamic range, the high sensitivity to X-rays and the usability in magnetic fields provide the basis for improved and new diagnostic and interventional methods. PMID- 12764580 TI - Hyperinsulinaemia and hyperglycaemia: possible risk factors of colorectal cancer among diabetic patients. AB - Hyperinsulinaemia and hyperglycaemia are two possible risk factors for colorectal cancer, which constitutes the third leading cause of cancer death in Western countries. Molecular evidence as well as animal models provide support for these associations: Insulin has been shown to be an important growth factor for colonic carcinoma cells, and both insulin and insulin-like growth factor-1 receptors have been detected in colon cancer tissue. The insulin-signal transduction pathway is involved in the regulation of gene expression and apoptosis. The role of hyperglycaemia in carcinogenesis could include pathways via luminal factors (related to fecal bile acid concentrations, stool bulk, and prolonged transit time) or circulatory factors (via glucose as the only energy source for neoplastic cells). This review summarizes the epidemiologic literature with respect to hyperinsulinaemia and hyperglycaemia as risk factors for colorectal cancer, and aims to integrate the biological and epidemiological evidence. Epidemiologic findings to date indicate a slightly increased risk of colorectal cancer for diabetic patients; however, there are some inconsistencies. Possible explanations for these inconsistencies include inadequate information about patients' diabetic disease and treatment states. We suggest that future studies should take medical history, staging and treatment for hyperinsulinaemia and hyperglycaemia into account to further our understanding of the role of hyperglycaemia and hyperinsulinaemia in colorectal carcinogenesis. PMID- 12764583 TI - [Digital thorax radiography: flat-panel detector or storage phosphor plates]. AB - Flat panel detectors are characterized by improved handling and increased dose efficiency. This allows for increasing of work flow efficiency and for reducing the exposure dose by about 50% compared to current systems with a sensitivity of 400. Whether the increased dose efficiency should be used to reduce acquisition dose or to increase image quality in the chest, will be shown by further clinical experience and will be also determined by the subjective preference of the radiologists. The decreased level of image noise opens new perspectives for image processing that way that elaborated multifrequency processing allows for optimizing the display of very small and low contrast structures that was so far limited by overlying image noise. Specialized applications of dual energy subtraction and temporal subtraction will also profit by the new detector technology and will be further driven forward in context with applications such as computed assisted diagnosis even though this is currently not yet broadly applied. Storage phosphor radiography still represents an important alternative technique based on its larger flexibility with respect to equipment configuration, its broader application options in intensive care and emergency radiology and due to economic reasons. These facts are further underlined by the fact that image quality also in storage phosphor radiography could be constantly increased by improving detector technology and image processing and consequently has a high standard. PMID- 12764584 TI - [Flat-screen detector systems in skeletal radiology]. AB - Implementation of flat-panel detectors and digital integration of the technique instead of the use of conventional radiographs leads to a shortening of the work process. With flat-panel technology the image production process is shortened by more than 30%. Major advantages in the implementation of integrated RIS, PACS and flat-panel detector system are increases in quality because most mistakes in picture labeling can be avoided, easier handling without the need for cassettes, and the possibility of image post-processing. The diagnostic quality of the images in the field of musculoskeletal radiology is, in comparison to conventional radiographs, at least adequate and in most cases markedly improved with a marked reduction in radiation exposure of around 30-50%. With respect to the numerous advantages of the digital techniques and especially flat-panel technology there is a very high likelihood that conventional radiographs will be substituted in the coming years, even though the cost of the new technology is currently significantly higher compared to conventional systems. PMID- 12764585 TI - [New computed radiography technologies in digital radiography]. AB - Digital radiography (DR) has become integral to modern diagnostic radiology. One of the earliest forms of DR, computed radiography (CR) using storage phosphors, has established itself as the mainstay of DR-based diagnostic imaging over the past 20 years. More recently, flat-panel DR systems based on solid state X-ray detectors with integrated, large-area, active-matrix readout electronics are promising further improvements in clinical workflow and image quality. Despite CR's longevity, innovations continue to be made. New developments in CR screen technologies, like structured (needle) screens, and new scanner concepts based on line-at-a-time reading promise major improvements in image quality (comparable to that of flat-panel systems), system through-put and physical size, at a cost comparable to that of today's systems. Thus, despite the advent of flat-panel acquisition systems, there will still be an important role for CR in the foreseeable future. After a brief review of the current state of CR technology, this paper will explore several of these new CR developments and present some examples of their potential clinical impact. PMID- 12764586 TI - [The amorphous selenium based flat-panel detector. Clinical experiences]. AB - The possibility of acquiring digital radiographic images has been marked with a rapid development. The main advantages of such images are, above all, the electronic dispersal of images, the economical archiving and the computer supported post processing. The recently introduced large-area flat panel detectors offer, contrary to storage-phosphor-based computed radiography, a direct read-out modality. It must be emphasized that differences between direct and indirect detector technology exist,which we will discuss here. The working and advantages of the amorphous selenium based direct-conversion detectors will be examined in detail. Clinical cases will be presented based on a three-year experience with over 200,000 exposures with a detector chest unit. PMID- 12764587 TI - [The use of flat-panel detectors for CT imaging]. AB - Clinical CT has reached a very high performance level by now. The introduction of spiral scanning and of multirow detectors have allowed to image even large body sections in very short time and with isotropic, high spatial resolution of better than 1 mm. For further improvements with respect to detector technology the use of flat-panel detectors (FPD), which have been developed for radiographic applications, is currently under investigation. In this article we discuss the general demands on CT detectors and specifically the suitability of FPDs with respect to CT imaging. FPDs offer excellent performance for the imaging of high contrast structures with high spatial resolution.Low-contrast resolution and dose efficiency, however, do not yet reach the level of performance of dedicated CT detectors; temporal resolution is also limited. FPDs appear primarily suited for special applications in CT as for example 3D angiography or intraoperative imaging which also allows for improvements in workflow. For standard diagnostic CT they are not to be recommended at present, last but not least for dose reasons. The respective technical developments will have to be reassessed constantly in the future. The development of detector systems which are equally suited for radiography and CT constitutes an attractive goal. PMID- 12764588 TI - [Monitoring of irradiated brain metastases using MR perfusion imaging and 1H MR spectroscopy]. AB - PURPOSE: In follow-up examinations of irradiated brain metastases conventional contrast-enhanced morphological MR imaging is often unable to distinguish between transient radiation effects, radionecrosis,and tumor recurrence. To evaluate changes of relative cerebral blood flow (rCBF) in irradiated brain metastases arterial spin-labeling techniques (ASL) were applied and compared to the outcome of (1)H MR spectroscopy and spectroscopic imaging ((1)H MRS, SI). PATIENTS AND METHODS: In 2 patients follow-up examinations of irradiated brain metastases were performed on a 1.5-T tomograph (average single dose: 20 Gy/80% isodose). Relative CBF values of gray matter (GM), white matter (WM),and metastases (Met) were measured by means of the ASL techniques ITS-FAIR and Q2TIPS. (1)H MRS was performed with PRESS 1500/135. RESULTS: In both patients with initially hyperperfused metastases (Met/GM >1) the reduction of rCBF after stereotactic radiosurgery indicated response to treatment--even if the contrast-enhancing region increased--while increasing rCBF values indicated tumor progression. The findings were confirmed by (1)H MRS, SI and subsequent follow-up. CONCLUSION: The ASL techniques ITS-FAIR and Q2TIPS are able to monitor changes of rCBF in irradiated brain metastases. The two cases imply a possible role for ASL-MR perfusion imaging and (1)H MR spectroscopy in differentiating radiation effects from tumor progression. PMID- 12764589 TI - [Inflammatory diseases of the orbit. Part 2: Bulb, extraconal space, lacrimal gland, opti nerve]. PMID- 12764600 TI - Remarkably stereoselective photoinduced electron-transfer reaction between zinc myoglobin and optically active binaphthyl bisviologen. AB - We have designed and synthesized new optically active bisviologens ([BNMV](4+)) containing a binaphthyl moiety to examine the stereoselective photoinduced electron-transfer (ET) reactions with zinc-substituted myoglobin (ZnMb) by flash photolysis. The photoexcited triplet state of ZnMb, (3)(ZnMb)*, was successfully quenched by [BNMV](4+) ions to form the radical pair of a ZnMb cation (ZnMb(.+)) and a reduced viologen ([BNMV](.3+)), followed by a thermal ET reaction to the ground state. The rate constants ( k(q)) for the ET quenching at 25 degrees C were obtained as k(q)( R)=(2.9+/-0.2)x10(7) M(-1) s(-1) and k(q)( S)=(2.2+/ 0.2)x10(7) M(-1) s(-1), respectively. The ratio of k(q)( R)/ k(q)( S)=1.3 indicates that the ( R)-isomer of the chiral viologen preferentially quenches (3)(ZnMb)*. On the other hand, the rate constants ( k) for the thermal ET reaction from [BNMV](.3+) to ZnMb(-+) at 25 degrees C were k( R)=(1.2+/ 0.1)x10(8) M(-1) s(-1) and k( S)=(0.47+/-0.03)x10(8) M(-1) s(-1), respectively, and the ratio remarkably increased to k( R)/ k( S)=2.6. The activation parameters, Delta H(not equal) and Delta S(not equal), were determined from the kinetic measurements at various temperatures (10-30 degrees C) to understand the ET mechanisms. In the quenching reaction, the energy differences of Delta Delta H*(R- S) and T Delta Delta S*( R- S) at 25 degrees C were calculated to be -3.9+/ 1.6 and -3.3+/-0.2 kJ mol(-1), respectively, whereas Delta Delta H*( R-S)=7.7+/ 1.9 kJ mol(-1 )and T Delta Delta S*( R-S)=9.9+/-0.5 kJ mol(-1 )were found for the thermal ET reaction. Therefore, the thermal ET reaction to the ground state was proved to be dominated by the entropy term, and the large stereoselectivity may arise from the decrease in charge repulsion between donor and acceptor. PMID- 12764603 TI - A comparison of the thermodynamics of O[bond]O cleavage for dicopper complexes in enzymes and synthetic systems. AB - The preferred state, the peroxide Cu(2)(II,II) or the bis-mu-oxo Cu(2)(III,III) states, for oxygen-bridged copper dimers with nitrogen donors is reinvestigated. Experiments have indicated that for the enzymatic complexes with histidine ligands the peroxide state should be favored, at least for hemocyanin, while for the synthetic complexes with strained ligands the bis-mu-xo state should be intrinsically favored. The present B3LYP study essentially agrees with these results. The quite different results obtained in CASPT2 and some previous B3LYP studies for these systems are investigated and discussed. The conclusion, drawn in an earlier study, that the Cu(2)(III,III) state is an unlikely intermediate in the enzyme mechanisms of tyrosinase and catechol oxidase, still remains. PMID- 12764601 TI - Structure-function relationship of reduced cytochrome c probed by complete solution structure determination in 30% acetonitrile/water solution. AB - The complete solution structure of ferrocytochrome c in 30% acetonitrile/70% water has been determined using high-field 1D and 2D (1)H NMR methods and deposited in the Protein Data Bank with codes 1LC1 and 1LC2. This is the first time a complete solution protein structure has been determined for a protein in nonaqueous media. Ferrocyt c retains a native protein secondary structure (five alpha-helices and two omega loops) in 30% acetonitrile. H18 and M80 residues are the axial heme ligands, as in aqueous solution. Residues believed to be axial heme ligands in the alkaline-like conformers of ferricyt c, specifically H33 and K72, are positioned close to the heme iron. The orientations of both heme propionates are markedly different in 30% acetonitrile/70% water. Comparative structural analysis of reduced cyt c in 30% acetonitrile/70% water solution with cyt c in different environments has given new insight into the cyt c folding mechanism, the electron transfer pathway, and cell apoptosis. PMID- 12764602 TI - Reduced hybrid cluster proteins (HCP) from Desulfovibrio desulfuricans ATCC 27774 and Desulfovibrio vulgaris (Hildenborough): X-ray structures at high resolution using synchrotron radiation. AB - The hybrid cluster proteins from the sulfate reducing bacteria Desulfovibrio desulfuricans ATCC 27774 ( Dd) and Desulfovibrio vulgaris strain Hildenborough ( Dv) have been isolated and crystallized anaerobically. In each case, the protein has been reduced with dithionite and the crystal structure of the reduced form elucidated using X-ray synchrotron radiation techniques at 1.25 A and 1.55 A resolution for Dd and Dv, respectively. Although the overall structures of the proteins are unchanged upon reduction, there are significant changes at the hybrid cluster centres. These include significant movements in the position of the iron atom linked to the persulfide moiety in the oxidized as-isolated proteins and the sulfur atom of the persulfide itself. The nature of these changes is described and the implications with respect to the function of hybrid cluster proteins are discussed. PMID- 12764604 TI - A global assessment using PCR techniques of mycorrhizal fungal populations colonising Tithonia diversifolia. AB - Tithonia diversifolia (Mexican sunflower) is a shrub commonly used as a green manure crop in Central and South America, Asia and Africa as it accumulates high levels of phosphorus and other nutrients, even in depleted soils. In root samples collected from the global distribution of Tithonia, we examined the degree of mycorrhizal colonisation and estimated the families of associated arbuscular mycorrhizal (AM) fungi. No colonisation by ectomycorrhizas was found. The degree of colonisation by AM fungi was on average 40%, but ranged between 0 and 80%. No mycorrhizal colonisation was found in the samples collected from the Philippines or in one each of the Rwandan and Venezuelan samples. Throughout its global distribution (Costa Rica, Nicaragua, Indonesia, Honduras, Mexico, Kenya and Rwanda), Tithonia forms mainly associations with Glomaceae. Only in one location in Nicaragua were associations with another family ( Acaulosporaceae) found. PMID- 12764605 TI - Influence of colonisation by an arbuscular mycorrhizal fungus on the growth of seedlings of Banksia ericifolia (Proteaceae). AB - Tap and primary lateral roots of seedlings of the putatively non-mycorrhizal Banksia ericifolia became marginally colonised when grown in an established mycelium of an arbuscular mycorrhizal (AM) fungus in the laboratory. A similar degree of colonisation was found in seedlings from an open woodland. All colonies lacked arbuscules. Two factors influencing colonisation and associated growth of host plants were examined experimentally: concentration of P in the soil and organic energy associated with the fungus. While some inoculated seedlings were slightly smaller when colonised by AM fungi, the results were inconsistent and never statistically significant. Seedlings take up insignificant quantities of soil P during early growth, even in the presence of abundant added P. Though colonisation was minor in all cases, an existing mycelium, whether or not connected to a companion plant, slightly increased the amount of root of B. ericifolia colonised by an AM fungus. All seedlings grew slowly. Shoots were significantly larger than roots, until the initiation of proteoid roots which commenced at about 40 days after germination, with both relatively high and low P supply. PMID- 12764606 TI - Antibodies against the PER protein of Drosophila label neurons in the optic lobe, central brain, and thoracic ganglia of the crickets Teleogryllus commodus and Teleogryllus oceanicus. AB - We describe labeling of neurons in the central nervous system of two cricket species, Teleogryllus commodus and T. oceanicus, with both mono- and polyclonal antibodies against the PER protein. Western blots reveal that the monoclonal antibodies recognize a single protein with a molecular weight of approximately 94 kDa, i.e., similar to that of the PER protein of the moth, Anterea pernii. Neurons and their processes are labeled both in the optic lobes and in the central brain. Processes occur in the accessory medulla, the medulla, and proximal lamina, in the central complex, in the non-glomerular neuropil, and in the retrocerebral complex, suggesting that PER-containing neurons form a widely distributed network. Neurons and processes were also labeled in the meso- and metathoracic ganglia. Four to six PER-immunoreactive (ir) neurons with processes in the accessory medulla were double labeled by an antibody against pigment dispersion factor (PDF), a peptide that is implicated in circadian rhythmicity in Drosophila. In the central brain, projections of fibers labeled by the anti-PER and anti-PDF antibodies were mainly distinct, with overlap only in a few restricted regions. In most neurons, including those projecting into the accessory medulla, PER labeling was restricted to the cytoplasm and there was no indication of circadian variation in the intensity of staining. PMID- 12764607 TI - Type 4A cAMP-specific phosphodiesterase is stored in granules of human neutrophils and eosinophils. AB - Persistent elevations of cAMP levels are generally accompanied by an inhibition of granulocyte functions. Phosphodiesterases play a critical role in regulating intracellular levels of cAMP. The expression of three isoforms of type 4 cAMP specific phosphodiesterase (PDE4) in neutrophils suggests diversity of isoform localization and targeting in regulating cell function. The sites of cAMP regulation in granulocytes by the PDE4A isoform were investigated by immunoelectron microscopy. PDE4A was localized uniformly in all granule classes of eosinophils, but was restricted in neutrophils to a subset of myeloperoxidase (MPO)-containing granules that were round or elongated with a central crystalloid core. Granulocytes were stimulated with fMLP to investigate the sites of PDE4A targeting during cell activation. In neutrophils, fMLP induced a rapid (1 min) translocation of granules containing PDE4A to the plasmalemma, where some PDE4A and MPO were exocytosed. In these cells, PDE4A labeling within granules was focal and no longer homogeneous. While immunogold labeling of PDE4A was reduced after fMLP stimulation, staining of MPO-containing granules remained high. Extracellular release of PDE4A was also observed in eosinophils stimulated with fMLP. Morphometry revealed that Au labeling was significantly reduced within 1 min, and that there was a shift in PDE4A localization within eosinophil granules from the crystalline core to the matrix. Fluctuations of cAMP levels and ectoprotein kinase activity with PKA properties occur in blood under normal and pathological conditions. The exclusive localization of PDE4A within granules of neutrophils and eosinophils suggests that PDE4A may function to downregulate cAMP signaling at the cell membrane and/or in the extracellular space at the time of granule release. PMID- 12764608 TI - Secretory apparatus assessed by analysis of pancreatic secretory stress protein expression in a rat model of chronic pancreatitis. AB - Secretory stress proteins (SSP) are a family of proteins including isoforms of pancreatitis-associated protein (PAP) and pancreatic stone protein (PSP/reg). In vitro exposure to trypsin results in the formation of insoluble fibrillar structures. SSP are constitutively secreted into pancreatic juice at low levels. The WBN/Kob rat is a model for chronic pancreatitis, displaying focal inflammation, destruction of the parenchyma and changes in the architecture of the acinar cell; the synthesis and secretion of SSP are also increased. We have investigated the secretory apparatus by SSP immunohistochemistry at the light- and electron-microscopical (EM) levels. Immunocytochemistry of PSP/reg in Wistar control rats reveals low levels, with individual acinar cells exhibiting high immunoreactivity in zymogen granules. PAP is not detectable. In the WBN/Kob rat, PSP/reg and PAP immunoreactivity is markedly increased. Double immunofluorescence for PSP/reg and PAP I or II demonstrates that these proteins colocalize to the same cell. Acinar cells change their secretory architecture by fusion of zymogen granules and elongation of the fused organelles. The immunogold technique has demonstrated an increase of SSP in zymogen granules in WBN/Kob rats. PSP/reg positive zymogen granules fuse to form elongated structures with fibrillar contents. An extensive PSP/reg-positive fibrillar network is established in the cytosol. Extracellular fibrils have been observed in several ductules. Thus, SSP derived fibrils form concomitantly with acinar damage in the WBN/Kob rat. Based on the known tryptic cleavage site of SSP, the in vivo generation of fibrils is presumably the result of premature trypsin activation. PMID- 12764609 TI - Histidine-rich glycoprotein plus zinc reverses growth inhibition of vascular smooth muscle cells by heparin. AB - Vascular smooth muscle cell (SMC) hyperplasia is known to be an important component in the pathogenesis of arteriosclerosis and restenosis. Although heparin has been well recognized as the representative molecule suppressing SMC growth in vitro, attempts to use heparin as a therapeutic anti-restenosis drug have not favorably influenced the angiographic or clinical outcome after angioplasty in some clinical trials. In this study, we have examined the effect of histidine-rich glycoprotein (HRG), a relatively abundant serum glycoprotein (~100 micrograms/ml in human serum), on the growth inhibition of cultured vascular SMC by heparin. Vascular SMC growth was significantly inhibited by heparin, giving nearly 85% inhibition with 100 micrograms/ml heparin. HRG reversed heparin-induced SMC growth inhibition in a dose dependent manner; 75% restoration of cell growth was observed when 100 micrograms/ml of HRG was co added with 100 micrograms/ml heparin. Interestingly, micromolar concentrations of the zinc ion (0-10 microM), compatible with concentrations released from activated platelets, were found to enhance the restorative action of HRG. Western blot experiment demonstrated no significant amounts of the HRG moiety in fetal bovine serum, eliminating the possible contribution of contaminant HRG from culture media. These findings indicate that HRG, in combination with the zinc ion, plays a role in modulating the SMC growth response in pathophysiological states and explain the lack of success of heparin as a therapeutic anti restenosis drug in clinical trials. PMID- 12764610 TI - Apoptosis and phosphatidylserine-mediated recognition during the take-over phase of the colonial life-cycle in the ascidian Botryllus schlosseri. AB - Colonies of the ascidian Botryllus schlosseri undergo recurrent generation changes in which adult zooids are gradually resorbed and replaced by new blastogenic generations. During these periods, known as take-over phases, programmed cell death, which, on the basis of morphological analysis is ascribed to apoptosis, occurs widely in zooid tissues. In the present report, we re investigate cell death during the take-over process. Results confirm the occurrence of diffuse apoptosis, as evidenced by chromatin condensation, positivity to the TUNEL reaction and expression of phosphatidylserine on the outer leaflet of the plasma membrane. Apoptosis also occurs among haemocytes, and senescent blood cells are actively recognised and ingested by circulating professional phagocytes. Both phosphatidylserine and CD36, a component of the thrombospondin receptor, are involved in the recognition of apoptotic haemocytes, which fosters the idea that fundamental recognition mechanisms are well conserved throughout chordate evolution. PMID- 12764611 TI - A gene for a Class II DNA photolyase from Oryza sativa: cloning of the cDNA by dilution-amplification. AB - Ultraviolet radiation induces the formation of two classes of photoproducts in DNA-the cyclobutane pyrimidine dimer (CPD) and the pyrimidine [6-4] pyrimidone photoproduct (6-4 product). Many organisms produce enzymes, termed photolyases, which specifically bind to these lesions and split them via a UV-A/blue light dependent mechanism, thereby reversing the damage. These photolyases are specific for either CPDs or 6-4 products. Two classes of photolyases (class I and class II) repair CPDs. A gene that encodes a protein with class II CPD photolyase activity in vitro has been cloned from several plants including Arabidopsis thaliana, Cucumis sativus and Chlamydomonas reinhardtii. We report here the isolation of a homolog of this gene from rice (Oryza sativa), which was cloned on the basis of sequence similarity and PCR-based dilution-amplification. The cDNA comprises a very GC-rich (75%) 5; region, while the 3; portion has a GC content of 50%. This gene encodes a protein with CPD photolyase activity when expressed in E. coli. The CPD photolyase gene encodes at least two types of mRNA, formed by alternative splicing of exon 5. One of the mRNAs encodes an ORF for 506 amino acid residues, while the other is predicted to code for 364 amino acid residues. The two RNAs occur in about equal amounts in O. sativa cells. PMID- 12764613 TI - Parvicapsula spinachiae n.sp. (Myxozoa, Parvicapsulidae) in Spinachia spinachia (L.) (Teleostei, Gasterosteidae) from Denmark. AB - Parvicapsula spinachiae sp. nov. is described from Spinachia spinachia (L.) (Teleostei, Gasterosteidae) from the northern Oresund, Denmark. The subspherical disporous plasmodia occur in the renal tubules and urinary bladder; the myxospores were found only in the urinary bladder. The myxospores are ovoid when viewed frontally and curved when viewed laterally. Mean maximum length is 10 microm and mean maximum width 5 microm. They have two spherical polar capsules in the apex and irregular thickenings at the posterior end. The newly described species is compared with the ten known species of Parvicapsula. It is most similar to Parvicapsula unicornis Kabata, 1962 but differs from this species by lacking the prominent suture and the posterior horn-like projection. PMID- 12764612 TI - Intra-isolate variation of Cryptosporidium parvum small subunit ribosomal RNA genes from human hosts in England. AB - Cryptosporidium parvum is a protozoan parasite responsible for a number of water borne outbreaks of human gastrointestinal infection. The importance of this parasite for human health has led to several investigations into its epidemiology. In the present study, sequence and PCR-RFLP analyses were performed on cloned SSU rRNA genes from human and bovine C. parvum isolates. Each cloned gene was assigned a genotype using AseI restriction digestion profiles. Although sequence heterogeneity was observed, six of seven human isolates contained human genotype SSU rRNA genes and one human isolate contained bovine genotype genes. No intra-isolate variation was observed between the SSU rRNA genes analysed from a single bovine (Iowa) isolate. Intra-isolate variation did not occur at the Ase I restriction sites and therefore did not interfere with the assignment of C. parvum genotypes by RFLP. Mixed RFLP genotypes (human and bovine) were not observed in any of the C. parvum isolates analysed. PMID- 12764614 TI - Left and right contributions to the Xenopus heart: implications for asymmetric morphogenesis. AB - The left-right asymmetry of the vertebrate heart is evident in the topology of the heart loop, and in the dissimilar morphology of the left and right chambers. How left-right asymmetric gene expression patterns influence the development of these features is not understood, since the individual roles of the left and right sides of the embryo in heart looping or chamber morphogenesis have not been specifically defined. To this end, we have constructed a bilateral heart-specific fate map of the left and right contributions to the developing heart in the Xenopus embryo. Both the left and right sides contribute to the conoventricular segment of the heart loop; however, the left side contributes to the inner curvature and ventral face of the loop while the right side contributes to the outer curvature and dorsal aspect. In contrast, the left atrium is derived mainly from the original left side of the embryo, while the right atrium is derived primarily from the right side. A comparison of our fate map with the domain of expression of the left-right gene, Pitx2, in the left lateral plate mesoderm, reveals that this Pitx2-expressing region is fated to form the inner curvature of the heart loop, the left atrioventricular canal, and the dorsal aspect of the left atrium. We discuss the implications of these results for the role of left right asymmetric gene expression in heart looping and chamber morphogenesis. PMID- 12764615 TI - Identification and developmental expression of a macrophage stimulating 1/ hepatocyte growth factor-like 1 orthologue in the zebrafish. AB - The cytokine Macrophage Stimulating 1 (MST1/MSP/Hepatocyte Growth Factor-Like) is a ligand of the Met-related MST1-Receptor (MST1R/RON). Although MST1-deficient mice are viable, MST1R is essential in mice before gastrulation for implantation, and is a known oncogene in man. Here I report the identification, sequence, chromosomal location and embryonic expression of a novel zebrafish orthologue, termed macrophage stimulating 1 (mst1). mst1 shows a striking restriction of expression to the dorsal side of the embryo prior to gastrulation, and as gastrulation and somitogenesis proceed is expressed sequentially in the presumptive neurectoderm, the notochord, the somites, endodermal cells and in the syncytial yolk. This dynamic pattern is largely conserved in tetrapod vertebrates, suggesting that the appearance of MST1, may have played an early role in the evolution of the vertebrate body plan. PMID- 12764617 TI - Morbidity among municipal waste incinerator workers: a cross-sectional study. AB - OBJECTIVES: In order that the occupational exposures and health risks for municipal waste incinerator operators be better determined, a study was carried out in three centres. METHODS: A transversal study was carried out for 102 male incinerator workers matched for age with 94 male workers from other industrial activities. Three groups of exposed workers were considered (group 1: "crane operators" and "equipment operators"; group 2: "furnace " workers; group 3: "maintenance" and "effluent-treatment" workers). Data were collected through a questionnaire and medical examination. The respiratory function was explored and blood samples were taken for biological analysis. Statistical analysis using chi(2) or Student's test, and calculation of odds-ratio (OR) using logistic regression analysis were performed. RESULTS: There were no significant differences in general symptoms between the two groups. Skin symptoms (OR =4.85; 2.04-11.51 for the "maintenance and effluent" workers) were more often observed in the exposed group. An excess of respiratory problems was also encountered: daily coughing ["maintenance and effluent" groups (OR =2.55; 0.84-7.75); "furnace men" (OR =6.58; 2.18-19.85)]. A significant relationship between exposure and the decrease of several pulmonary parameters was observed. The liver and haematological tests were well within the normal range; nevertheless, a slight but significant increase in the white blood cells was observed in the exposed group. Higher blood lead levels were found for exposed workers. CONCLUSION: There were few adverse health effects in our morbidity study but skin irritation and cough were more frequent in the exposed workers. A slight decrease in pulmonary function was observed. These findings are in agreement with those of several other studies. PMID- 12764616 TI - Neuronal endoplasmic reticulum acts as a single functional Ca2+ store shared by ryanodine and inositol-1,4,5-trisphosphate receptors as revealed by intra-ER [Ca2+] recordings in single rat sensory neurones. AB - We addressed the fundamentally important question of functional continuity of endoplasmic reticulum (ER) Ca(2+) store in nerve cells. In cultured rat dorsal root ganglion neurones we measured dynamic changes in free Ca(2+) concentration within the ER lumen ([Ca(2+)](L)) in response to activation of inositol-1,4,5 trisphosphate receptors (InsP(3)Rs) and ryanodine receptors (RyRs). We found that both receptors co-exist in these neurones and their activation results in Ca(2+) release from the ER as judged by a decrease in [Ca(2+)](L). Depletion of Ca(2+) stores following an inhibition of sarco(endoplasmic)reticulum Ca(2+)-ATPase by thapsigargin or cyclopiazonic acid completely eliminated Ca(2+) release via both InsP(3)Rs and RyRs. Similarly, when the store was depleted by continuous activation of InsP(3)Rs, activation of RyRs (by caffeine or 0.5 microM ryanodine) failed to produce Ca(2+) release, and vice versa, when the stores were depleted by activators of RyRs, the InsP(3)-induced Ca(2+) release disappeared. We conclude that in mammalian neurones InsP(3)Rs and RyRs share the common continuous Ca(2+) pool associated with ER. PMID- 12764620 TI - Electrode insertion depth in cochlear implantees estimated during surgery, on plain film radiographs and with electrode function testing. AB - Three methods of determining electrode insertion depth in cochlear implantees are studied: intraoperative counting of inserted electrodes, plain film radiography using Stenvers projection and postoperative electrode function testing. In 16 cases the number of electrodes inserted in the cochlea were counted both by the surgeon at surgery and by two independent observers on plain film radiographs using Stenvers projections. The electrode function was tested postoperatively. The differences between the three methods in estimation of the number of intracochlear electrodes were analyzed with t-tests, and 95% confidence intervals (95% CI) of the mean differences were calculated. The mean difference between the radiograph observers was 0.25 electrode (95% CI, -0.69 to 1.19 electrodes.) The mean difference between radiography observations and the surgical counts was 0.60 electrode (95% CI, -0.71 to 1.91 electrodes.) The mean difference between surgical counting and electrode function testing was 0.40 electrode (95% CI, 0.66 to 1.46 electrodes.) The mean difference between radiograph observations and electrode function testing was 0.50 electrode (95% CI, -0.51 to 1.51 electrodes.) No significant differences existed between the three methods. Our findings showed similar results in estimating electrode array insertion depth with the three methods. Plain film radiography using Stenvers projection is satisfactory if imaging is indicated for determining the number of inserted electrodes. PMID- 12764618 TI - Immunofluorescent detection of activation of initiator caspases-8 and -9 during pharmacologically induced apoptosis of cultured HeLa and endothelial cells. AB - Caspases, a group of cysteine-activated aspartate-directed proteases, play an integral role in the execution of programmed cell death or apoptosis. In the cellular caspase cascade, the processing of native proenzymes into activated forms of downstream, effector caspases is dependent on the activation of initiator caspases-8 and -9. We describe a staining procedure for immunofluorescence-based analysis of activation of caspase-8 and -9 during pharmacologically induced apoptosis in primary cultures of human umbilical vein derived endothelial cells and in an established line of HeLa cells. Using cleavage site-directed antibodies, specific intracellular detection for cleaved fragments of caspase-8 and -9 was accomplished during apoptosis induced by staurosporine and etoposide. The population of cells displaying morphological signs of apoptosis, evidence for DNA strand breaks by TUNEL analysis, and positive staining for active forms of caspase-8 and caspase-9 increased with the duration of treatment, suggesting activation of initiator caspases in correlation with the onset and progression of apoptosis. The application of immunocytochemical staining procedures for quick and specific in situ detection may effectively aid the identification of participating upstream caspases and elucidation of complex apoptosis signaling mechanisms. PMID- 12764621 TI - Papillary carcinoma of the thyroglossal duct cyst in childhood. AB - Thyroglossal duct carcinoma is a rare malignancy that is usually diagnosed postoperatively. Approximately 150 cases have been reported in the literature. Eighty-five percent of these were papillary carcinomas. Controversies exist concerning its nature and treatment. In this report, we present an 11-year-old boy with an anterior cervical cystic mass originating in the thyroglossal duct. After a primary Sistrunk procedure, the cyst and tract extending to the foramen caecum at the base of the tongue in continuity with the midportion of the hyoid bone were resected. Histopathologic study demonstrated a papillary carcinoma. After 4 months of follow-up, the patient is asymptomatic without any evidence of recurrence. The clinical and histopathological features and therapeutic options are discussed. PMID- 12764622 TI - Vacuum application through a nonfully dilated cervix: a viable option. AB - The aim of this cross-sectional study was to assess the feasibility of vacuum delivery through a nonfully dilated cervix. The study group consisted of 39 women with vacuum deliveries through a nonfully dilated cervix larger than 9 cm and station of the head at S or more +2 cm. These were compared to a control group of 215 vacuum deliveries at a fully dilated cervix and 46 women who underwent cesarean section at a nonfully dilated cervix larger than 9 cm. The main indication for vacuum extraction in the study group was fetal distress and in the control groups prolonged 2nd stage, dysfunctional labor and fetal distress. Maternal and neonatal morbidity was low and not different between the groups. Neonatal well being, evaluated by cord pH and 5-min Apgar score, was not different. Based on predefined criteria, vacuum extraction through a nonfully dilated cervix is a viable alternative to emergency cesarean section and is apparently not associated with higher maternal or infant morbidity. PMID- 12764623 TI - Women's perception of transvaginal sonography in the first trimester; in an early pregnancy assessment unit. AB - The objective of this study was to gauge women views of transvaginal sonography in an emergency setting and to study any correlation between their perception of the procedure and their prior knowledge and experience of it. We surveyed women presenting with various complications of the first trimester. Only women who had transvaginal sonography and who gave informed consent were included. Four hundred and twenty-five women were recruited. Transvaginal sonography was considered not embarrassing, acceptable, not painful and not stressful by 378 (88.9%), 417 (98.1%), 419 (98.6%) and 385 (90.6%) of the women respectively. There were no statistically significant differences in the women's perception of the procedure whether they had prior knowledge and previous experience of the procedure or not. The majority of the women perceived transvaginal sonography favourably. Their perception of the procedure was favourable whether they had prior knowledge and previous experience of it or not. PMID- 12764624 TI - The effect of metformin treatment to ovarian response in cases with PCOS. AB - OBJECTIVE: To examine the effect of metformin therapy on ovarian response in clomiphene-resistant PCOS patients. DESIGN: Randomized prospective study. SETTING: Zeynep Kamil Hospital IVF Center. PATIENTS: Thirty-two female PCOS patients with clomiphene citrate resistance. INTERVENTIONS: Test group A received 2x850 mg/day metformin therapy for 8 weeks, followed by gonadotropin induction. Only gonadotropin induction was applied to Control group B. MAIN OUTCOME MEASURES: Total gonadotropin dosage, duration of gonadotropin therapy, estradiol level on HCG-day, number of follicles with >or=16 mm diameter, number of cases with hyperstimulation development, number of cancelled cycles, endometrial thickness on HCG-day, pregnancy outcome, multiple pregnancy rate. RESULTS: The total gonadotropin doses given to metformin-study group was significantly lower than the control group. In addition, duration of therapy, HCG level, plasma estradiol level on HCG-day in the study group was significantly lower than in the control group. CONCLUSIONS: Metformin therapy has led to a higher pregnancy outcome, as well as to a decrease in hyperstimulation risk, cycle cancellations, and multiple pregnancy rates with a lower gonadotropin dosage. PMID- 12764625 TI - Internal jugular vein thrombosis as a late complication of ovarian hyperstimulation syndrome in an ICSI patient. AB - BACKGROUND: Thromboembolism is a rare but extremely serious complication of ovarian hyperstimulation syndrome. CASE REPORT: We report a case of left internal jugular vein thrombosis, presenting as a late complication of moderate ovarian hyperstimulation syndrome. CONCLUSION: Prevention of vascular thrombosis should be a part of the treatment in the setting of assisted reproductive procedures complicated with ovarian hyperstimulation syndrome. PMID- 12764626 TI - Laparoscopic management of cornual pregnancy without sutures. AB - INTRODUCTION: Cornual pregnancy is a rare form of ectopic pregnancy. The incidence is reported as 3% of all ectopic pregnancies accounting 20% of deaths due to ectopic pregnancy. When an unruptured cornual pregnancy is diagnosed, there are a variety of management options. Many successful endoscopic management options for cornual pregnancy have been reported. CASE REPORT: In this case, cornual resection was performed. DISCUSSION: The other possible treatment options were reviewed. PMID- 12764627 TI - Neuropathological and behavioral changes induced by various treatment paradigms with MPTP and 3-nitropropionic acid in mice: towards a model of striatonigral degeneration (multiple system atrophy). AB - We characterized two models of dual nigral and striatal lesions replicating the lesion pattern of striatonigral degeneration, the neuropathological hallmark of parkinsonism associated with multiple system atrophy (SND/MSA-P). For this purpose, we used systemic administration of 1-methyl-4-phenyl-1, 2, 3, 6 tetrahydropyridine (MPTP) and 3-nitropropionic acid (3-NP) in C57BL mice. One group of animals was first injected with MPTP followed by 3NP (MPTP+3-NP model). In the second group 3-NP was injected first, followed by MPTP (3-NP+MPTP model). The behavioral and neuropathological characteristics of these two models were compared to those observed after single 3-NP or MPTP intoxication. Results showed that, compared to control mice, spontaneous nocturnal locomotor activity was preserved in the MPTP+3-NP model, whereas it was reduced by 27% ( P<0.05) in the 3-NP+MPTP model and in animals treated with either 3-NP (27%, P<0.05) or MPTP (23%, P<0.05) alone. Quantitative histological evaluation based on Nissl staining and DARPP-32 immunohistochemistry revealed that 3-NP alone and 3-NP+MPTP treatment produced a marked (greater than 50%) loss of striatal neurons, whereas MPTP+3-NP treatment attenuated loss of striatal neurons by 43%. Further, loss of tyrosine hydroxylase-positive neurons in substantia nigra pars compacta (SNc) was attenuated after 3-NP+MPTP treatment compared to that observed after MPTP (40% vs 74%, P<0.001) and MPTP+3NP treatment (55% vs 74%, P<0.01). Our results show that MPTP-induced nigral lesions attenuate 3-NP toxicity and, reciprocally, that 3-NP induced striatal lesions reduce MPTP toxicity. This suggests that complex integrative mechanisms are likely to regulate the vulnerability of the striatum and SNc to cell death in SND/MSA-P. PMID- 12764628 TI - T-cell-mediated mucosal immunity is attenuated in experimental necrotizing enterocolitis. AB - In premature infants with necrotizing enterocolitis (NEC) the gastrointestinal mucosal barrier is immature, but little is known about the immune response of immature bowel. The aim of this study was to evaluate the intestinal mucosal immune response in experimental NEC. In general anaesthesia, NEC was induced in six newborn piglets by injection of bovine casein into terminal ileum. Six controls received an equal amount of saline. Four hours later, samples were taken from the macroscopically most affected part of the treated loop and from the macroscopically healthy untreated intestine. Monoclonal antibodies to porcine CD1, CD2, CD4, CD8, CD45 and IgM were used for immunohistochemical staining. Casein-treated bowel showed typical macro- and microscopic findings of NEC. No changes were found in the saline-treated bowel. In both groups the bowel outside the treatment sector was normal. In casein-treated animals, treated samples showed significant decrease in density of CD4+ cells when compared with saline treated controls. Similar trend was found in CD2+ and CD8+ cells but without statistical significance. Macroscopically healthy proximal untreated samples showed significant decrease in densities of CD2+, CD4+ and CD8+ lymphocytes in casein-treated group when compared with control samples. In casein-treated animals the density of CD45+ cells in the non-injected bowel was also decreased, but this did not reach statistical significance. Densities of CD1+ and IgM+ cells did not differ between casein-treated and saline-treated groups. A significant T cell decrease was found in the present NEC model. Surprisingly, this was most prominent in the macroscopically healthy bowel outside the casein injection segment. The reason for T-cell decrease remains unclear, but bovine casein is known to contain peptide fractions that can modulate immune function. These findings may have implications in the design of neonatal milk formulas. PMID- 12764629 TI - Long-term outcome of hydrocephalus management in myelomeningoceles. AB - BACKGROUND: The cerebrospinal fluid (CSF) shunt remains an important ongoing management problem in myelomeningocele (MMC) patients. We reviewed the long-term shunt treatment outcome in a prospectively followed group of MMC patients from a single institution. METHOD: Patients prospectively entered into a hydrocephalus database with a diagnosis of MMC from the years 1987 to 1996 were selected. All data was verified from the medical records and additional details about the shunt surgery were collected. The outcome of shunt failure was categorized as shunt obstruction, shunt infection, presence of loculated ventricles, overdrainage, and other. All deaths were recorded and causation identified. Univariate analysis for shunt failure risk factors was accomplished using Log rank statistics. Multivariable analysis was performed for each repeated failure level using a conditional Cox regression model. RESULTS: One hundred and twenty (64%) out of 189 MMC patients experienced a first shunt failure with a median time of 303 days; 29 (24%) of the failures were due to shunt infection (the procedure infection rate being 15%). Sixty-one patients experienced a second shunt failure, 38 a third and 36 had four or more. Multivariable analysis of risk factors failed to demonstrate any clear risk factors for either first or subsequent shunt failure. Fifteen patients died, 13 from either shunt or Chiari 1 complications. CONCLUSION: Shunt complications remain an important cause of morbidity and mortality in MMC patients, particularly shunt infection. PMID- 12764630 TI - Defining torpor in free-ranging bats: experimental evaluation of external temperature-sensitive radiotransmitters and the concept of active temperature. AB - A variety of definitions involving body temperature (Tb), metabolic rate and behavior have been used to define torpor in mammals and birds. This problem is confounded in some studies of free-ranging animals that employ only skin temperature (Tsk), a measure that approximates but may not precisely reflect Tb. We assess the accuracy of Tsk in the context of a recent definition for torpor called active temperature. We compared the active temperatures of individual big brown bats (Eptesicus fuscus), which aggregate in cavities, with solitary, foliage-roosting hoary bats (Lasiurus cinereus). In captive big brown bats, we compared Tsk and core Tb at a range of ambient temperatures for clustered and solitary roosting animals, compared Tsk and Tb during arousal from torpor, and quantified the effect of flight on warming from torpor. Hoary bats had significantly lower active temperatures than big brown bats despite having the same normothermic Tsk. Tsk was significantly lower than Tb during normothermia but often greater than Tb during torpor. Flight increased the rate of warming from torpor. This effect was more pronounced for Tsk than Tb. This suggests that bats could rely on heat generated by flight muscles to complete the final stages of arousal. Using active temperature to define torpor may underestimate torpor due to ambient cooling of external transmitters or animals leaving roosts while still torpid. Conversely, active temperature may also overestimate shallow torpor use if it is recorded during active arousal when shivering and non-shivering thermogenesis warm external transmitters. Our findings illuminate the need for laboratory studies that quantify the relationship between metabolic rate and Tsk over a range of ambient temperatures. PMID- 12764632 TI - Abstracts of the European Society of Gastrointestinal and Abdominal Radiology (ESGAR) 14th Annual Meeting. June 17-21, 2003. Budapest, Hungary. PMID- 12764631 TI - Distinct load dependence of relaxation rate and diastolic function in Oryctolagus cuniculus and Ratus norvegicus. AB - This study investigated potential differences on load dependence of relaxation rate and diastolic function between Oryctolagus cuniculus and Ratus norvegicus, which have constitutive differences in the mechanisms involved in myocardial inactivation. Load dependence of relaxation rate and diastolic function were evaluated with the response of left ventricular time constant tau and diastolic pressure-dimension relation to beat-to-beat aortic constrictions in open-chest rabbits and rats. Afterload levels were normalized, being expressed as a percentage of peak isovolumetric pressure (relative load). In control heartbeats, relaxation rate and diastolic function were similar in the two animal species. They presented, however, distinct responses to afterload elevations. In rabbits, time constant decreased approximately 7% and diastolic pressure-dimension relation remained unchanged when afterload was elevated to a relative load of 73 76%. Above this afterload level, a significant deceleration of relaxation rate (increase of time constant) and an upward shift of diastolic pressure-dimension relation were observed. In rats, afterload elevations accelerated pressure fall up to a relative load of 97-100% and no afterload-induced shift of the diastolic pressure-dimension relation was observed. This study provides, therefore, evidence that Oryctolagus cuniculus has lower afterload reserve of myocardial relaxation and diastolic function than Ratus norvegicus. PMID- 12764634 TI - Comparison of lesion size estimated by dynamic MR imaging, mammography and histopathology in breast neoplasms. AB - To evaluate the accordance of size measurements of malignant breast lesions 65 women with 76 malignant lesions were preoperatively examined with triple diagnosis (mammography was performed in three views with additional views if necessary) and dynamic MR imaging using a subtraction technique with a 3D T1 weighted sequence. Maximum lesion size at histopathology was used as gold standard and compared with maximum lesion size at MRI and mammography. All measurements were made independently for each method. Histopathology verified 48 invasive, 5 in situ, and 23 mixed lesions. No significant difference was found for the pure invasive lesions ( p=0.366). In the mixed lesions a slightly better result for MRI was indicated ( p=0.116), although there was a great spread. Only five pure in situ lesions were assessed, too few to draw any statistical conclusions ( p>0.5). An overall difference indicated a slight superiority of MRI ( p=0.097). The MR imaging and mammography are both good at measuring the size of detected invasive breast malignancies. The total sizes of mixed lesions are frequently underestimated by both MRI and mammography, although the invasive parts were equally well described and measured with both methods. PMID- 12764636 TI - Lymphoma: pictorial review. II. AB - Lymphoma is a spectrum of malignant neoplasms of the lymphoid system. The appearances are radiologically diverse with almost all organs susceptible to involvement. The condition is challenging as it can mimic the appearance of almost all other neoplasms. This review article highlights in a general way the diversity of the radiological appearances and organs. PMID- 12764635 TI - Accuracy of tumor size measurement in breast cancer using MRI is influenced by histological regression induced by neoadjuvant chemotherapy. AB - The aim of this study was to evaluate whether regressive changes after neoadjuvant chemotherapy for breast cancer affect the accuracy of preoperative MRI measurements of tumor size. Thirty-one patients with breast cancer underwent MRI before and after neoadjuvant treatment. Besides pre- and post-contrast T1 weighted MRI, dynamic MRI with high temporal resolution (turbo-FLASH) was performed. Contrast enhancement in dynamic MRI was quantified using a pharmacokinetic two-compartment model, where two parameters, amplitude and k(ep), were calculated and color coded on transversal parameter maps. Considering the conventional MR images, tumor diameters were measured on the color maps and compared with histological tumor size. Histological regression was scored on a five-point scale regarding cytopathic effects, reactive changes, and tumor cell reduction. The correlation between tumor sizes measured by MRI and histopathology was 0.83 ( p<0.0007) in 12 tumors without regressive changes (score 0), and 0.48 ( p<0.051) in 17 tumors with regressive changes scored 1 or 2, without any tendency for systematic over- or underestimation. In two cases without residual tumor (score 4), MRI likewise showed no signs of persistent tumor. The decrease of the contrast enhancement parameters was significantly more marked in tumors with signs of histological regression than in those without. Whenever MRI is used to judge the response of breast cancer to chemotherapy, the reader must be aware that therapy-induced changes may cause significant over- or underestimation of tumor size. We saw a high precision only when there was either no response - according to histological criteria - or when the tumor had regressed completely. PMID- 12764637 TI - On visual determination of full inspiration on CT images. AB - The aim of this study was to evaluate the ability of experienced thoracic radiologists to assess full inspiration based on two CT slices, one above and one below the carina, in normal subjects. Ten healthy volunteers were studied. Total lung capacity (TLC) was measured with a body plethysmograph. High-resolution computed tomography (HRCT) was performed in two slices at TLC and at various expired volumes. Mean Hounsfield values (HU) were calculated. Unidentifiable images, stored on a web server, were analysed visually by experienced thoracic radiologists. The results show that the mean lung density at TLC varied by approximately 40 HU between individuals. Within an individual this may correspond to a decrease in lung volume of approximately 25% of TLC. On visual determination of images taken at 65-74% of TLC, more than one-third of the images were assessed as taken at full inspiration; of the images taken at 75-84% of TLC, approximately 50% were assessed as taken at full inspiration. We conclude that visual determination of full inspiration on CT images in normal subjects is highly inaccurate. If quantitative density measurements are to be used in the diagnosis or follow-up of lung disease, thorough control of full inspiration is recommended. PMID- 12764638 TI - Low-dose multislice CT of the thorax in follow-up of malignant lymphoma and extrapulmonary primary tumors. AB - Our objective was to study the impact of low-dose multislice thoracic CT on image quality and lesion conspicuity in follow-up of patients with malignant lymphoma and extrapulmonary primary tumors. Forty consecutive patients with a history of malignant lymphoma or extrapulmonary malignant primaries who had undergone previous standard-dose thoracic spiral CT (120 kVp, 150 mAs, 8 mm) were subjected to low-dose multislice CT (15 mAs/rotation, 120 kVp, pitch 1.5, 4x2-mm collimation reconstructed to contiguous 8-mm slices. Image quality and lesion conspicuity were classified independently by two readers on a four-point ordinal scale (1=poor, 4=excellent). Mean image quality was significantly lower for low dose CT, 2.35, than for standard-dose CT, 3.25 ( p<0.001); however, all low-dose CT examinations were fully readable, none had to be repeated, and no lesions (including 51 lung lesions and 31 soft tissue lesions) remained undetected. Only in the apical lung did the lowered tube current significantly reduce lesion conspicuity. Even in pulmonary lesions smaller than 10 mm ( n=21) lesion conspicuity did not significantly differ between standard and low-dose scans. Low dose thoracic multislice CT based on a 90% reduction in dose compared with standard-dose techniques was not associated with impaired detection of suspicious lesions and may be useful in follow-up of malignant lymphoma and extrapulmonary tumors. PMID- 12764639 TI - X-ray imaging of the chest in Switzerland in 1998: a nationwide survey. AB - The use in Switzerland of three chest X-ray imaging modalities is studied. The frequency and dosimetric aspects associated with chest radiography, chest CT, and photofluorography are investigated and the Swiss figures are compared with those of other countries. The frequencies are established by means of a nationwide survey performed in 1998, whereas the doses are determined by modelling, based on the technical parameters of the examinations. The annual frequencies of chest radiography, chest CT, and photofluorography are 201, 7, and 7 per thousand population, respectively. The standard effective doses related to these three types of examinations are 0.057, 9, and 0.13 mSv, respectively. The collective dose associated with chest X-ray imaging amounts to 606 man.Sv, representing 8.5% of the total collective dose due to diagnostic and interventional radiology. The geographic and seasonal variations, as well as the distribution with the medical specialty and with the age of the patient, are established. The Swiss results compare well with the average data published in the literature for other countries of similar health care level. Chest radiography shows a significant decrease with time. It dropped by a factor of 2 in 20 years. The conformity of the indication of chest X-ray modalities with referral guidelines is also investigated. PMID- 12764640 TI - The use of ECG and respiratory triggering to improve the sensitivity of oxygen enhanced proton MRI of lung ventilation. AB - Changes in lung signal between normal breathing and breathing pure oxygen is the basis for oxygen-enhanced ventilation MRI. An optimal technique guarantees a significant response to pure oxygen in well-ventilated lung tissue. To improve the sensitivity, we investigated the effect of ECG and respiratory triggering. Centric reordered single-shot rapid acquisition relaxation enhancement sequences (TE 4.2 ms, echo spacing 4.2 ms, bandwidth 650 Hz/pixel), with an inversion recovery preparation pulse (TI 700 ms), were used. Series of 20 measurements were performed with and without ECG and respiratory triggering in five young volunteers. Subsequently, series of 100 images were acquired during breathing normal air and pure oxygen (as "stimulus"). Ventilation maps showed by means of the z-score how far the response deviates from the signal intensities during the normal air condition. The standard deviation of the lung signal intensities was lowest for the cardiac-triggered series. In the ventilation maps, on the other hand, signal changes were statistically more significant in the respiratory than in the cardiac-triggered series. The average z-scores in the right (left) cranial part of the lung were 12.4 (13.0) and 9.2 (9.7) for respiratory and ECG-triggered acquisitions. We propose to use respiratory triggering as a means to improve the sensitivity of MR ventilation studies. PMID- 12764641 TI - Ferucarbotran (Resovist): a new clinically approved RES-specific contrast agent for contrast-enhanced MRI of the liver: properties, clinical development, and applications. AB - Ferucarbotran (Resovist) is the second clinically approved superparamagnetic iron oxide developed for contrast-enhanced MRI of the liver. The purpose of this review is to provide an overview on the properties, clinical development, and application of ferucarbotran. Safety data obtained during clinical phases I-III revealed a total of 162 adverse events within 1053 patients, of which 75 were classified as possibly, probably, or definitely drug related. The majority of events occurred within the first 3 h (73 of 75) and was of mild intensity. The agent significantly improves the detection of hypovascular focal liver lesions with a comparable sensitivity in lesion detection to CTAP but without a relevant loss in specificity. Furthermore, ferucarbotran leads to a significant improvement of the sensitivity for lesion classification and characterization of the most frequent liver lesions. Contrast-enhanced MRA is not feasible and the angiographic effect is not sufficient to allow for postprocessing of data into maximum intensity projections. Intraindividual studies at low-field (0.2 T) and high-field (1.5 T) showed similar rates for lesion detection. The time window for contrast-enhanced MRI of the liver is at least 1 day up to 4 days. The compound can be regarded as safe and well tolerated. Even bolus injections caused no cardiovascular side effects, lumbar back pain, or clinically relevant laboratory changes. The examination time can be kept short with T1- and T2-weighted pre contrast sequences, dynamic MRI over 10 min, and finally accumulation phase T2 weighted MRI. Patients who may benefit in particular are surgical candidates for resection, transplantation, or interventional therapies, and patients with liver cirrhosis and/or suspected hepatocellular carcinoma to either exclude malignancy or to define the extent of disease, the location of lesions, and the type of newly detected lesions. PMID- 12764643 TI - Assessment of organ volume with different techniques using a living liver model. AB - The purpose of this study was to compare different techniques for the estimation of liver volume, and to evaluate errors associated with volume estimation techniques based on linear measurements. Fifteen patients with focal liver lesion underwent spiral CTA for preoperative evaluation. The scan protocol was: collimation 3 mm; rotation time 0.75 s; pitch 2; and reconstruction index 1 mm. Reconstructed images were sent to a workstation running on a NT platform equipped with post-processing software allowing 3D reconstructions. Linear measurements and volume estimation through manual segmentation were obtained with preset window and magnification. Volume was calculated from linear measures using different equations. With equations based on linear measurements the right lobe was overestimated (mean=+53%; mean error=14.7%), the second and third segments were underestimated (mean=-47%; mean error=43.3%) and the total volume was underestimated (mean=-86%; mean error=36%). The volume calculated by summing the areas of all the sections and multiplying the result by the increment was not significantly different from the volume estimated using the manual volumetric segmentation technique. Volume estimation obtained through linear measurements is not reliable to appraise the volume of irregular-shaped solids, even assuming the prevalence of a particular morphology. PMID- 12764642 TI - 3D contrast-enhanced MR portography and direct X-ray portography: a correlation study. AB - Our objective was to compare 3D contrast-enhanced MR portography (3D CE MRP) on a 1.5-T MR imager with direct X-ray portography. Twenty-six consecutive patients underwent 3D CE MRP with in-plane resolution of 1.4 or 1.8 mm, and direct X-ray portography. The findings of these two methods were evaluated and compared. The main portal vein (PV), right PV with its anterior and posterior segmental branches, and left PV including its sagittal segment were shown clearly without diagnostic problem in all cases on MRP. The main PV appearance was accordant with MRP and X-ray. For intrahepatic PVs, the results agreed in 21 patients but disagreed in 5 patients. In 1 patient with a huge tumor in right liver, the right posterior PV was classified as occluded at MRP, but diffusely narrowed at X-ray. The findings of left intrahepatic PV were discordant in 3 patients with hepatocelluar carcinoma in the left lobe. The MRP demonstrated complete occlusion of the left PVs, whereas X-ray showed proximal narrowing and distal occlusion. In another patient with hepatocelluar carcinoma, a small non-occlusive thrombus involving the sagittal segment of the left PV was seen on MRP but not on X-ray. With demonstration of varices and portosystemic shunts, MRP showed results similar to those of X-ray, except one recanalized para-umbilical vein was excluded from the field of view at MRP due to the patient's limited ability of breathholding. The 3D CE MRP correlated well with direct X-ray portography in most cases, it was limited in distinguishing narrowing of an intrahepatic PV from occlusion, but it showed advantage in demonstrating small thrombus within PV. PMID- 12764644 TI - Image distortion in US-guided liver tumor puncture with curved linear array. AB - Our objective was to systematically examine the influence of probe geometry, puncture route, and composition of the abdominal wall on the deviation error with needle display. We made a simplified simulation model of the puncture needle passing to the liver tumor, using a microcomputer system, and visualized the degree of distortion of the puncture needle, changing the thickness and component of the abdominal wall as well as the radius of the probe, site, and angle of puncture. The puncture needle exhibited a displacement error of -10.9 to 3.8 mm (center hole) and -11.3 to 3.8 mm (side attachment). The shift was less accentuated when the radius of the probe was large. The displacement error was less accentuated in the center and accentuated in the left side of the field of view. It is recommended that liver puncture under US guidance be generally performed using a probe with a larger radius through the center hole with the most obtuse puncture angle possible, but when the tumor is localized in the hepatic dome, a more tightly curved probe should be used with an acute angle. We must consider a possible large error in display of the puncture needle during liver puncture and try to prevent hazardous complications. PMID- 12764645 TI - Feasibility of ultra-low-dose multislice CT colonography for the detection of colorectal lesions: preliminary experience. AB - Our objective was to evaluate the feasibility of ultra-low-dose scanning for multislice CT colonography in the detection of colorectal lesions. Twenty-seven patients (14 men, 13 women) with clinical indication for conventional colonoscopy were recruited. Multislice spiral CT (Somatom Plus 4 Volume Zoom, Siemens, Germany) examinations were performed after standard oral colonoscopic preparation and colonic distension with room air. Images were acquired using 2.5-mm collimation, 3.0-mm slice thickness, standard reconstruction kernel, 140 kVp, and 10 mAs. Supine and prone acquisitions were obtained in all patients. Images were analyzed on a workstation by two gastrointestinal radiologists. Conventional colonoscopy was performed on the same day in all patients and represented the standard of reference. Total radiation exposure was also calculated. All colorectal cancers were correctly identified at CT colonography (9 of 9, sensitivity 100%). The CT colonography also detected 10 of 12 polyps (overall sensitivity 83.3%). Based on polyp diameter, the sensitivity for the detection of polyps 10 mm or larger was 100% (3 of 3); between 6 and 9 mm, 100% (3 of 3); and 5 mm or smaller, 66.6% (4 of 6). Total radiation exposure for prone and supine acquisitions combined was 1.7 mSv (for men) and 2.3 mSv (for women). Although preliminary, our experience suggests that ultra-low-dose scanning for multislice CT colonography is feasible. This technique provides a sensitivity comparable to that of previous experiences with CT colonography but with a 40-70% reduction of the radiation dose delivered to patients. PMID- 12764646 TI - Prospective comparison of MR enteroclysis with multidetector spiral-CT enteroclysis: interobserver agreement and sensitivity by means of "sign-by-sign" correlation. AB - Our objective was a prospective comparison of MR enteroclysis (MRE) with multidetector spiral-CT enteroclysis (MSCTE). Fifty patients with various suspected small bowel diseases were investigated by MSCTE and MRE. The MSCTE was performed using slices of 2.5 mm, immediately followed by MRE, obtaining T1- and T2-weighted sequences, including gadolinium-enhanced acquisition with fat saturation. Three radiologists independently evaluated MSCTE and MRE searching for 12 pathological signs. Interobserver agreement was calculated. Sensitivities and specificities resulted from comparison with pathological results ( n=29) and patient's clinical evolution ( n=21). Most pathological signs, such as bowel wall thickening (BWT), bowel wall enhancement (BWE) and lymphadenopathy (ADP), showed better interobserver agreement on MSCTE than on MRE (BWT: 0.65 vs 0.48; BWE: 0.51 vs 0.37; ADP: 0.52 vs 0.15). Sensitivity of MSCTE was higher than that of MRE in detecting BWT (88.9 vs 60%), BWE (78.6 vs 55.5%) and ADP (63.8 vs 14.3%). Wilcoxon signed-rank test revealed significantly better sensitivity of MSCTE than that of MRE for each observer ( p=0.028, p=0.046, p=0.028, respectively). Taking the given study design into account, MSCTE provides better sensitivity in detecting lesions of the small bowel than MRE, with higher interobserver agreement. PMID- 12764647 TI - Detection of intestinal drug containers by ultrasound scanning: an airport screening tool? AB - Our objective was to investigate the value of abdominal ultrasound for the detection of intestinal drug containers in air travelers suspected of intestinal drug trafficking. The study population consisted of 50 suspects charged with intestinal drug smuggling. All suspects, 7 women and 43 men, underwent an abdominal ultrasound scan immediately after detention in the prison hospital. A simple portable ultrasound unit (SonoSite 180) was used with a 2- to 4-MHz curved array transducer. The images were digitally stored or recorded onto videotape. All suspects remained at the prison hospital to have their stool monitored for drug containers. The Customs officers who determined the presence or absence of containers in the stools were unaware of the ultrasound examination results. Of the suspects, 42 (84%) proved to have multiple intestinal drug containers as assessed by stool examination during detention. In 40 of 42 suspects abdominal ultrasound correctly identified the presence of containers. In 7 of 8 suspects ultrasound correctly identified the absence of containers. The positive predictive value of an ultrasound examination was 97.6%. In this group of suspects an abdominal ultrasound provided accurate information about the presence or absence of intestinal drug containers in 47 of 50 cases (94%). PMID- 12764648 TI - Performance of a flat-panel detector in the detection of artificial erosive changes: comparison with conventional screen-film and storage-phosphor radiography. AB - The purpose of this study was to compare a large-area, direct-readout, flat-panel detector system with a conventional screen-film system, a storage-phosphor system, and a mammography screen-film system with regard to the detection of artificial bone erosions simulating rheumatoid disease, and to assess its diagnostic performance with decreasing exposure dose. Six hundred forty regions were defined in 160 metacarpophalangeal and proximal interphalangeal joint specimens from 20 monkey paws (4 regions per joint). Artificial bone erosions were created in 320 of these 640 regions. Specimens were enclosed in containers filled with water to obtain absorption and scatter radiation conditions similar to those of a human hand. Imaging was performed using a flat-panel system, a speed class 200 screen-film system, a mammography screen-film system, and a storage-phosphor system under exactly matched conditions. Different exposure doses equivalent to speed classes of S=100, 200, 400, 800, 1600, and 3200 were used. In all images the presence or absence of a lesion was assessed by three radiologists using a five-level confidence scale. Receiver operating characteristic (ROC) analysis was performed for a total of 21,120 observations (1920 for each imaging modality and exposure level) and diagnostic performance estimated by the area under the ROC curve (A(z)). The significance of differences in diagnostic performance was tested with analysis of variance. The ROC analysis showed A(z) values of 0.809 (S=200), 0.768 (S=400), 0.737 (S=800), 0.710 (S=1600), and 0.685 (S=3200) for the flat-panel system, 0.770 for the speed class 200 screen-film system, 0.781 (S=200), 0.739 (S=400), 0.724 (S=800), 0.680 (S=1600) for the storage-phosphor system, and 0.798 for the mammography screen film system. Analysis of variance showed significant differences between different combinations of imaging modalities and exposure doses ( p<0.05). The diagnostic performance of the flat-panel detector system is superior to that of a screen-film system and a storage-phosphor system for the detection of erosive lesions at clinical exposure settings (S=200). Using the flat-panel system the exposure dose can be reduced by 50% to obtain a diagnostic performance comparable to a speed class 200 screen-film system. PMID- 12764649 TI - Open radio-frequency thermal ablation of renal VX2 tumors in a rabbit model using a cooled-tip electrode: feasibility, safety, and effectiveness. AB - The purpose of this study was to evaluate the feasibility, safety, and effectiveness of radio-frequency (RF) ablation using an internally cooled-tip electrode on renal VX2 tumors implanted in rabbits. Thirty-three rabbits with implanted renal VX2 tumors were divided into two groups: an RF ablation (RFA) group ( n=27) and a control group ( n=6). In the RFA group, RFA was performed on 27 implanted VX2 tumors using a cooled RF electrode and they were divided into three subgroups according to the follow-up period: acute (1-3 days, n=12); subacute (1-4 weeks, n=9); and chronic (2-7 months, n=6). Contrast-enhanced spiral CT was performed before the RFA and at the day, day 3, weeks 1, 2, 4, and months 2 and 7, after the RFA. The therapeutic efficacy was evaluated by the survival rate, CT, and pathologic findings. The RFA of renal tumors was technically successful in each instance. Complete tumor ablation was achieved in 22 of the 27 rabbits (81.5%) in the RFA group: 5 rabbits survived longer than 8 weeks without any evidence of viable tumor (18.5%) and 17 rabbits were found free of viable tumors when killed (63.0%). Five rabbits showed local tumor relapse and/or hematogenous lung metastasis after ablation (a recurrence rate of 18.5%). There were 11 (40.7%) complications related to the procedure. This experimental study demonstrates the feasibility of RFA therapy to treat renal VX2 tumors in rabbits, although RFA for central tumors carries some major potential complications, including renal arterial injury. PMID- 12764650 TI - Hemolytic effect of deformed intra-arterial stents and stent grafts in vitro. AB - Our objective was to evaluate the hemolytic effect of stents and stent grafts in an in vitro flow model. The model consisted of silicone tubings. Pulsatile flow was delivered at 1170 ml/min. Diluted packed erythrocytes were used (hematocrit of 33%). The Palmaz, MegaLink stents or Passager, Hemobahn stent grafts were placed in the middle portion of the model; control experiments contained no implant ( n=10 each). Concentric stenosis of the implant was achieved by constricting the implant to 25, 50, or 70% (area, n=10 each). Samples were drawn at minute 1, 2, 4, 6, 8, 10, 15, and every 10 min up to 1 h. Hemolytic parameters were evaluated. All implants produced hemolysis, increasing in the course of time. Without deformation (0%) the Hemobahn stent causes less hemolysis compared with the other prostheses. At 25% no significant difference could be shown between all devices. At 50% constriction the Palmaz stent caused more hemolysis than the other grafts, and at 70% stenosis the Palmaz and Hemobahn caused more hemolysis than Passager or Megalink. A hemolytic effect was evaluable, probably due to mechanical stress and sheer forces, induced by turbulences in the proximity of the deformed stent. The stent grafts did not perform better than the stents. Differences in the design of a prosthesis and in the degree of stent deformation seem to have an influence on the hemolysis caused. PMID- 12764651 TI - Sonographic evaluation of gluteus medius and minimus tendinopathy. AB - Our objective was to describe the sonographic appearance of the gluteus medius and minimus tendons in normal subjects, and to illustrate the spectrum of sonographic findings in gluteus tendinopathies. Sonography was performed in 20 asymptomatic volunteers. Seventy-five consecutive patients (59 women, 16 men; mean age 57.1 years) presented with pain and point tenderness over the greater trochanter. There were 43 right hips and 32 left hips. Ten patients provided a history of a traumatic incident with subsequent symptoms (mean duration 3.2 months). All patients underwent sonography to assess the site and severity of injury, and to discriminate tendinosis from partial and complete tear. Calcific foci, bony change, and fluid in the trochanteric bursae were noted. Twenty-two patients subsequently underwent surgery. Fifty-three (53 of 75) patients showed sonographic evidence of gluteus medius tendinopathy. Twenty-eight patients were thought to have tendinopathy without discrete tear. Sixteen patients had partial tears and 9 full-thickness tears. Gluteus minimus tendinopathy was detected in 10 of 75 patients. Foci of tendinopathy and partial tears were more common in the deep and anterior portions of the gluteus medius tendon attachment. Eight patients had fluid pooling in the trochanteric bursae. Findings were confirmed in 22 patients at surgery. Sonography can identify gluteus medius and minimus tendinopathy and provides information about the severity of the disease. PMID- 12764652 TI - MR evaluation of bone cysts treated with intracavital steroid injection. AB - The objective of this study was to evaluate the role of MR examination in showing the effects of intralesional steroid injection on bone cysts. Nineteen patients with radiologically suspected (group A, n=15) or already treated (group B, n=4) bone cysts underwent MRI (0.5 T; protocol: T2* gradient recalled echo; short tau inversion recovery or inversion recovery fast spin echo; T1 SE before and after contrast administration, 0.2 mmol/kg). All patients underwent steroid therapy for bone cysts and were followed up for 6-18 months after treatment. Evaluation of bone cyst response to therapy was based mainly on the detection of reparative tissue outlining all cystic portions at first MRI control. In group A, as early as 1 month after the first injection, the presence of a thin reparative tissue lining the cystic wall was observed. Follow-up studies revealed the progressive thickening of this reparative tissue and new bone formation. Similar features were present in the cysts of group B. Residual cystic cavities were depicted in 7 cases, with no evidence of enhancing tissue, requiring further treatment. The formation of a reparative tissue is the first feature of bone cyst recovery after intracavital steroid injection and is appreciable only on MRI. Because of the optimal visualisation of this reparative tissue and of residual cystic cavities, MRI could be proposed as a non-invasive, alternative and effective tool for evaluating the efficacy of steroid injection therapy of bone cysts. PMID- 12764653 TI - Impingement syndrome of the ankle following supination external rotation trauma: MR imaging findings with arthroscopic correlation. AB - Our objective was to identify MR imaging findings in patients with syndesmotic soft tissue impingement of the ankle and to investigate the reliability of these imaging characteristics to predict syndesmotic soft tissue impingement syndromes of the ankle. Twenty-one ankles with chronic pain ultimately proven to have anterior soft tissue impingement syndrome were examined by MR imaging during January 1996 to June 2001. The MR imaging protocol included sagittal and coronal short tau inversion recovery (STIR), sagittal T1-weighted spin echo, axial and coronal proton-density, and T2-weighted spin-echo sequences. Nineteen ankles that underwent MR imaging during the same period of time and that had arthroscopically proven diagnosis different than impingement syndrome served as a control group. Fibrovascular scar formations distinct from the syndesmotic ligaments possibly related to syndesmotic soft tissue impingement were recorded. Arthroscopy was performed subsequently in all patients and was considered the gold standard. The statistical analysis revealed an overall frequency of scarred syndesmotic ligaments of 70% in the group with ankle impingement. Fibrovascular scar formations distinct from the syndesmotic ligaments presented with low signal intensity on T1-weighted images and remained low to intermediate in signal intensity on T2-weighted MR imaging. Compared with arthroscopy, MR imaging revealed a sensitivity of 89%, a specificity of 100%, and a diagnostic accuracy of 93% for scarred syndesmotic ligaments. The frequency of scar formation distinct from the syndesmotic ligaments in patients with impingement syndrome of the ankle was not statistically significantly higher than in the control group. In contrast to that, anterior tibial osteophytes and talar osteophytes were statistically significantly higher in the group with anterior impingement than in the control group. Conventional MR imaging was found to be insensitive for the diagnosis of syndesmotic soft tissue impingement of the ankle. Fibrovascular scar tissue distinct from syndesmotic ligaments is suggestive for the diagnosis of soft tissue impingement, but the reliability of these findings is still questionable. PMID- 12764654 TI - Lymphadenopathy at the medial epitrochlear region in cat-scratch disease. AB - Our objective was to describe the imaging features of lymphadenopathy at the medial epitrochlear region as frequently seen in cat-scratch disease (CSD) and to compare these features with those seen in non-CSD. Ten patients (four males and six females) presented with a subcutaneous soft tissue mass at the medial epitrochlear region. Three patients were younger than 15 years. Magnetic resonance imaging was performed in all patients. All lesions turned out to be regional lymphadenopathy instead of soft tissue neoplasm. They were caused by benign disorders in eight cases, including seven CSD and one other infection, and in two cases by metastatic disease. All lesions, except for three benign ones, were solitary. On MRI, high signal intensity (SI) strands in the surrounding soft tissue on T2-weighted images (WI) were present in all cases. Three cases of CSD showed homogeneous SI on both T1- and T2WI. After contrast administration, a moderate, homogeneous enhancement was seen. The other seven lesions, except for one metastasis, had both solid and liquid components. Liquid components showed high SI on T2WI without enhancement, whereas solid components had intermediate SI on T2WI and marked enhancement after contrast administration. Pathological findings in seven patients, including four with CSD, supported the tentative diagnosis made on MRI. One of two patients with CSD had positive serological test. Cat-scratch disease is a major cause of a soft tissue swelling at the medial epitrochlear region. Identification of lymphadenopathy and its presentation on MRI may be helpful in eliciting an animal-scratch history and achieving diagnostic and therapeutic aims in patients without primary cancer. PMID- 12764655 TI - MR findings in knee osteoarthritis. AB - Knee osteoarthritis (OA) is a leading cause of disability. Recent advances in drug discovery techniques and improvements in understanding the pathophysiology of osteoarthritic disorders have resulted in an unprecedented number of new therapeutic agents. Of all imaging modalities, radiography has been the most widely used for the diagnosis and management of the progression of knee OA. Magnetic resonance imaging is a relatively recent technique and its applications to osteoarthritis have been limited. Compared with conventional radiography, MR imaging offers unparalleled discrimination among articular soft tissues by directly visualizing all components of the knee joint simultaneously and therefore allowing the knee joint to be evaluated as a whole organ. In this article we present the MR findings in knee OA including cartilage abnormalities, osteophytes, bone edema, subarticular cysts, bone attrition, meniscal tears, ligament abnormalities, synovial thickening, joint effusion, intra-articular loose bodies, and periarticular cysts. PMID- 12764656 TI - MRI assessment of right ventricular dysplasia. AB - Right ventricular dysplasia is a new entity of unknown origin in the classification of cardiomyopathies. Also known as arrhythmogenic right ventricular cardiomyopathy (ARVC) or arrhythmogenic right ventricular dysplasia, it is a disease of the heart muscle characterised by fibroadipose atrophy mainly involving the right ventricle and responsible for severe ventricular arrhythmias and sudden death also in young people. Magnetic resonance imaging provides evidence of ventricular dilatation at the outflow tract, thinning and thickening of the wall, diastolic bulging areas (especially located at the level of the right ventricle outflow tract) and fatty substitution of the myocardium mainly at the level of the right ventricle. Many radiologists erroneously consider the previously described fatty substitution as the main sign of ARVC, even though an evaluation of fat substitution alone may be a source of error for two reasons: firstly, because isolated areas of fatty replacement are not synonymous with ARVC since small non-transmural focal fatty areas of fat are also present in the normal patients; and secondly, because the MRI detection of fat may be overestimated due to partial-volume artefacts with normal subepicardial fat. Cardiac MRI can also be employed for the diagnosis of idiopathic right ventricular outflow tract tachycardia. Considering the evolutive nature of the disease, the non-invasiveness of MRI allows the follow-up of these patients and may be considered an excellent screening modality for the diagnosis of ARVC in family members. Finally, MRI can be employed in electrophysiological studies to locate the arrhythmogenic focus and reduce sampling errors. PMID- 12764657 TI - Effect of partial left ventriculectomy on left and right ventricular volumes and function as assessed with electron beam tomography: preliminary results. AB - Our objective was to determine if left ventricular reduction surgery affects left and right ventricular volumes and function. Twenty-three patients with end-stage heart failure underwent contrast-enhanced electron beam tomographic function studies before and twice after partial left ventriculectomy (PLV). The PLV was combined with other cardiosurgical procedures in 20 of 23 patients. Left and right ventricular enddiastolic volume (LVEDV, RVEDV), stroke volume (LVSV, RVSV), ejection fraction (LVEF, RVEF), and enddiastolic diameter (LVEDD, RVEDD) were determined by manual tracing of endo- and epicardial borders at enddiastole and endsystole. Patients were scanned 31 days (+/-34) before and 18 days (+/-13) and 8 months (+/-4) after PLV. Mean pre- and early and late postoperative values for LVEDV, LVSV, LVEF, and LVEDD were 387.9 ml (+/-125.5 ml), 255.6 ml (+/-79.3 ml; p<0.01), and 253.7 ml (+/-97.8 ml; p<0.05), 79.7 ml (+/-25.2 ml), 74.8 ml (+/ 17.9; n.s.), and 79.1 ml (+/-26.5 ml; n.s.), 21.6% (+/-7.3%), 31.9% (+/-13.4%; p<0.05), and 34.1% (+/-14.1%; p<0.05), and 72.0 mm (+/-10.6 mm), 64.3 mm (+/-8.5 mm; p<0.05), and 63.5 mm (+/-9.4 mm; p<0.05), respectively. Mean pre- and postoperative values for RVEDV, RVSV, RVEF, and RVEDD were 177.7 ml (+/-72.8 ml), 172.4 ml (+/-59.2 ml; n.s.), and 178.9 ml (+/-60.8 ml; n.s.), 60.3 ml (+/-21.6 ml), 68.8 ml (+/-19.9 ml; n.s.), and 78.3 ml (+/-25.3 ml; n.s.), 38.1% (+/ 15.4%), 43.7% (+/-16.3%; p<0.05), and 45.1% (+/-11.2%; n.s.), and 50.4 mm (+/ 10.9 mm), 48.1 mm (+/-8.7 mm; n.s.), and 48.5 mm (+/-9.8 mm; n.s.), respectively. The PLV may induce a significant early reduction of left ventricular volumes and improvement of biventricular function; however, our results must be judged carefully as the majority of patients in this study underwent additional cardiosurgical procedures, the contributory effect of which on the overall outcome remains unclear. PMID- 12764658 TI - Characterization of renal tumours with pulse inversion harmonic imaging by intermittent high mechanical index technique: initial results. AB - Our objective was to evaluate whether contrast-specific ultrasound (US) technique pulse inversion harmonic imaging (PIHI) with Levovist could reveal differential morphological features in solid and cystic renal masses. Twenty-six renal masses in 26 patients were evaluated by PIHI after Levovist injection with intermittent high mechanical index stimulation, performed every 10-15 s during arterial and delayed phase. Helical CT (HCT; n=6) or histological findings on surgical/bioptic specimen ( n=20) were considered as the reference procedures for definitive diagnosis. Eleven solid renal cell carcinomas (RCCs), 1 embryonal matanephric adenoma (EMA), 7 angiomyolipomas (AMLs), 4 cystic RCCs and 3 complex inflammatory cysts were identified. Solid RCCs revealed a much higher ( p<0.05) contrast enhancement than AMLs with typical pattern on conventional US ( n=6). The EMA and AML with atypical pattern on conventional US ( n=1) revealed intense contrast enhancement during arterial phase, progressively decreasing during delayed phase. Cystic RCCs and complex inflammatory cysts revealed intense contrast enhancement on the peripheral thick wall during arterial phase decreasing on delayed phase. The PIHI has been shown to differentiate solid RCC from typical AML by contrast enhancement assessment after Levovist injection, but differential diagnosis both in solid and in cystic renal masses still remains difficult. PMID- 12764659 TI - Duplex ultrasound as first-line screening test for patients suspected of renal artery stenosis: prospective evaluation in high-risk group. AB - Our objective was to assess how far the progress in ultrasound devices has increased feasibility and accuracy of Duplex ultrasound (DUS) for the diagnosis of renal artery stenosis (RAS), in a population with high prevalence of atherosclerotic renovascular lesions. Ninety-one hypertensive patients with atherosclerotic disease were prospectively evaluated by both DUS and digital subtraction angiography (DSA) of the renal arteries. Only proximal criteria (peak systolic velocity >180 mm/s or renal-to-aortic ratio >3.5) were used for the diagnosis of significant RAS (>60% narrowing). For both techniques, two readers were involved for interobserver variability study. Two hundred one arteries were demonstrated by DSA on 182 available kidneys. The prevalence of RAS among the study group was 37%. Sixteen of the 19 accessory arteries were not seen at DUS; in 8 patients, one renal artery was not seen at DUS (feasibility 91%). On the 177 arteries assessed, in comparison with DSA, DUS yielded 96, 91, and 97% mean values of accuracy, sensitivity, and specificity, respectively. Kappa for interobserver agreement was 0.95 for DUS and 0.92 for DSA. Although still unreliable for the detection of accessory arteries, DUS is in our experience an accurate and reproducible diagnostic test for RAS. PMID- 12764660 TI - A new device for retrograde urethrography in the male. AB - A new technique is presented for retrograde urethrography (RGU) using a plastic adjustable clamp. The procedure was performed in 36 patients with suspected urethral stenosis. Mean age was 55.3 years. The device and the procedure are described. All the radiologic examinations of the urethra were carried out successfully. The urethrograms showed images of stenosis in 27 patients. The stenoses localized in the distal tip of the anterior urethra were clearly seen with this clamp system. This method enables evaluation of the cases with stenosis close to the meatus that are difficult or even impossible to visualize with the conventional method. When necessary, a finer catheter can be used. This clamp system is recommended for RGU. The technique is simple, effective, and well tolerated by patients. PMID- 12764661 TI - Pediatric excretory MR urography: comparative study of enhanced and non-enhanced techniques. AB - Our purpose was to compare the quality of ureteral imaging in pediatric patients using two different MR sequences: the non-enhanced heavily T2-weighted (W) turbo spin-echo sequence (TSE) and the gadolinium-enhanced T1W fast-field-echo sequence (T1 FFE). An experimental study on three pigs was first performed. The TSE, before and after furosemide injection, was followed by the T1 FFE sequence. The clinical study included 11 infants and 10 children. With some modifications the same MR parameters and techniques were used as in the animal study. The TSE with TR 8000 ms and TE 650 ms implied 6 radial stacks each of 40 mm thickness. The T1 FFE included TR 18 ms, TE 2.9 ms, flip angle 60, and 50 slices with thickness 0.7 mm. After post-processing, image reconstructions qualitative and quantitative analysis were performed. Complete visualization of the ureters was achieved in 35 of 42 (83%) cases. Seventy-four percent of the ureters were completely visualized with T1 FFE compared with only 19% with TSE. Sixty-nine percent of the ureters were better imaged with T1 FFE than TSE and 21% equally well imaged. Four ureters (10%), either obstructed or due to poor renal function, were better imaged with TSE. The two sequences are complementary. Visualization of non-obstructed ureters is excellent with T1 FFE and the sequence is superior to TSE. The TSE, however, may be equal to or even better than T1 FFE in visualizing obstructed ureters or ureters draining malfunctioning kidneys. PMID- 12764662 TI - Ultrasound of the appendix in children: is the child too obese? AB - The negative influence of obesity on the detection rate of the appendix for US in adults has been reported. It has been assumed that obesity is a limiting factor in the detection of the appendix with US in children as well, but this has not yet been proven. The aim of our study was to evaluate whether nutritional condition (defined by the body mass index-for-age percentiles: BMI-FAP) influences the detection of the appendix in children on US. One hundred twenty six children (65 girls and 61 boys) with a mean age of 11.4 years with clinically suspected acute appendicitis underwent ultrasound on a commercially available high-end machine (HDI 5000, ATL, Bothell, Wash.). The BMI was calculated, and children were divided in three weight groups in accordance with the BMI-FAP, and were correlated with US findings. Evaluation of the three weight groups in accordance with the BMI-FAP demonstrated significant differences ( p=0.04) in the detection of the appendix. There was no statistical significance for the BMI, weight, height, and age solely for the detection of the appendix. In children there is a correlation between the nutritional condition as defined by the BMI FAP and the detection of the appendix. PMID- 12764663 TI - CSF flow imaging in the management of third ventriculostomy with a reversed fast imaging with steady-state precession sequence. AB - The aim of this study was evaluation of a fast and slow-flow sensitive 2D steady state free-precession sequence for its capability to prove the patency of endoscopic third ventriculostomy (TVS) in obstructive hydrocephalus, and to exclude communicating third ventricle prior to TVS. We compared gated and ungated variants of this sequence for this purpose. Twenty-three patients with obstructive hydrocephalus underwent 36 MR examinations with a 2D reversed fast imaging with steady-state precession (PSIF) sequence in a retrospectively cardiac gated (cine) and a faster but ungated version beside T1- and T2-weighted sequences in three planes. Thirteen patients were examined both before and after TVS, 4 patients solely before, and 6 patients solely after TVS. Imaging diagnoses were compared with intraoperative findings and clinical findings after TVS. Preoperative diagnosis of non-communicating third ventricle and cisterns was intraoperatively confirmed in 16 of 17 cases. Preoperative MRI was inconclusive in 1 case. Postoperative MRI revealed sufficient TVS in 16 of 19 cases and obstructed TVS in 3 of 19 cases due to several reasons. Findings at MRI were consistent in 19 of 19 cases with the clinical course and intraoperative results. The faster but ungated PSIF sequence was found to be diagnostically equivalent to the cardiac gated cine sequence. The CSF flow imaging with a 2D reversed fast imaging with steady-state precession sequence in conjunction with conventional T1 and T2-weighted images is a fast and reliable tool for pre- and postoperative functional evaluation in third ventriculostomy. PMID- 12764664 TI - Application of multi-detector row helical scanning to postmyelographic CT. AB - The advent of a multi-detector row helical CT has made it possible to attain images over a broader area with good spatial resolution. We assessed whether postmyelographic CT scans obtained using this system provided more information than conventional imaging techniques. Postmyelographic CT scans were preoperatively obtained in 46 patients using a multi-detector row helical CT system. Reconstructed images in the sagittal and coronal planes in all patients and curved reformation images along the dural sac in 37 patients were compared with myelograms and conventional postmyelographic CT scans. In 34 patients comparison was also made with MR images. The multi-detector row CT images demonstrated deformities of the dural sac more clearly than the other modalities in 39 of the 46 patients. They also provided the best visualization of nerve root abnormalities in 24 of the 46 patients and clearly revealed the presence of spurs in all 22 patients with spinal canal stenosis. Postmyelographic CT scans made using a multi-detector row helical CT system provide more information on the dural sac, nerve sleeves, and their contents than other imaging techniques. PMID- 12764665 TI - Posterior semicircular canal dehiscence: a morphologic cause of vertigo similar to superior semicircular canal dehiscence. AB - The aim of this study was to assess imaging findings of posterior semicircular dehiscence on computed tomography and to evaluate incidence of posterior and superior semicircular canal dehiscence in patients presenting with vertigo, sensorineuronal hearing loss or in a control group without symptoms related to the inner ear. Computed tomography was performed in 507 patients presenting either with vertigo ( n=128; 23 of these patients suffered also from sensorineuronal hearing loss), other symptoms related to the inner ear, such as hearing loss or tinnitus ( n=183) or symptoms unrelated to the labyrinth ( n=196). All images were reviewed for presence of dehiscence of the bone, overlying the semicircular canals. Twenty-nine patients had superior semicircular canal dehiscence. Of these patients, 83% presented with vertigo, 10% with hearing loss or tinnitus and the remaining 7% with symptoms unrelated to the inner ear. In 23 patients dehiscence of the posterior semicircular canal was encountered. Of these patients, 86% presented with vertigo, 9% with hearing loss or tinnitus and 5% with symptoms unrelated to the inner ear. Defects of the bony overly are found at the posterior semicircular canal, in addition to the recently introduced superior canal dehiscence syndrome. Significant prevalence of vertigo in these patients suggests that posterior semicircular canal dehiscence can cause vertigo, similar to superior semicircular canal dehiscence. PMID- 12764666 TI - Morpho-functional evaluation of osteomeatal complex in chronic sinusitis by coronal CT. AB - The objective was to determine whether or not there exists a clinical radiological correlation in chronic sinusitis of anterior paranasal sinus and to quantitatively evaluate the importance of osteomeatal complex structures and anatomic variants in chronic sinusitis etiopathology. We analyzed 208 nasal fossae with high-resolution and low-irradiation coronal computed tomography. The following inclusion criteria we established: one case control and one control group. We considered any chronic thickness of sinusal mucosal as a radiological indicator of chronic sinusitis. We evaluated the mucosal thickness of internal, external, superior and inferior maxillary walls, the addition of four maxillary walls and the maximal mucosal thickness in frontal sinus and anterior ethmoidal cells. We also obtained 15 parameters of osteomeatal complex structures and anatomical variants in each nasal fossa, and we correlated them statistically with chronic sinusitis radiological indicators. Uncinate process length is the only anatomic element from which we have been able to obtain a statistically significant cut point between case and control groups, but only with a sensitivity of 51.6%, specificity of 71.2%, positive predictive value of 74.2%, and negative predictive value of 47.9%. No association of radiological parameters exists that can explain chronic sinusitis. PMID- 12764667 TI - Suppressor mutations define two regions in the Cbp1 protein important for mitochondrial cytochrome b mRNA stability in Saccharomyces cerevisiae. AB - Nuclear-encoded Cbp1 stabilizes and promotes translation of mitochondrial cytochrome b (COB) mRNA. A CCG triplet within the 5'UTR of COB mRNA is essential for Cbp1-dependent stability. Like cbp1 mutations, mutation of any nucleotide in CCG results in degradation of COB transcripts. In this study, CBP1-linked pseudorevertants of the temperature-sensitive CCU strain were isolated. The suppressors are missense mutations within a central cluster or a carboxyl cluster in the linear sequence of Cbp1. Strains with mutations in the carboxyl half of the central cluster or the carboxyl cluster respire better than those with mutations in the amino half of the central cluster. COB mRNA levels in the suppressor strains were increased compared with that in the CCU strain and were positively correlated with respiratory capability. This correlation supports a model in which the primary role of Cbp1 is to protect COB mRNAs and deliver them to the mitochondrial translational apparatus. PMID- 12764669 TI - Meiotic behavior of a supernumerary chromosome in Magnaporthe oryzae. AB - A 1.2-Mb DNA band from an isolate of Magnaporthe oryzae was detected in a pulsed field gel. A chromosomal entity corresponding to this band was observed at the mitotic metaphase stage. This minichromosome, carrying many transposable elements and two telomeres, was transmitted to ascosporic F(1) cultures in a non-Mendelian manner with frequent changes in its size and number. Segregation analysis with RFLP markers indicated that the minichromosome underwent structural rearrangements, such as deletion and duplication, not only during meiosis but also after meiosis. An ectopic sister chromatid recombination may cause the size variation of the minichromosomes. PMID- 12764668 TI - The role of adaptor protein Ste50-dependent regulation of the MAPKKK Ste11 in multiple signalling pathways of yeast. AB - In Saccharomyces cerevisiae, Ste50 functions in cell signalling between the activated G protein and the mitogen-activated protein kinase (MAPK) kinase kinase (MAPKKK) Ste11. ScSte50 is an essential component of three MAPK-mediated signalling pathways, which control the mating response, invasive/filamentous growth and osmotolerance (HOG pathway), respectively. ScSte50 signalling may also contribute to cell wall integrity in vegetative cells. The protein contains a sterile alpha motif (SAM) and a putative Ras-associated domain (RAD), which are essential for signal transduction. Ste50 and Ste11 interact constitutively via their SAM regions. Ste50 interacts weakly and probably transiently with the pheromone receptor-bound heterotrimeric G protein G(alpha beta gamma), and with the small G proteins Cdc42, Ras1 and Ras2. It is specifically the RAD region of Ste50 that mediates the interactions with Cdc42 and Ras. Homologues of ScSTE50 are also found in other fungi, like S. kluyveri, Hansenula polymorpha, Candida albicans and Neurospora crassa. In this review, the role of Ste50 as an adaptor that links the G protein-associated Cdc42-Ste20 kinase complex to the effector kinase Ste11 and thus modulates signal transduction, especially in the pheromone response pathway of S. cerevisiae, is discussed. PMID- 12764670 TI - Clinical effects of combination therapy with mitoxantrone, vincristine, and prednisolone in breast cancer. AB - PURPOSE: We assessed the clinical efficacy and safety of mitoxantrone hydrochloride which has been used as an anticancer drug in our hospital to treat breast cancer patients since 1993. METHODS: A group of 23 patients with breast cancer were given one course of the following regimen every 3 weeks: mitoxantrone hydrochloride (8 mg/m(2) i.v. day 1), vincristine sulfate (1.2 mg/m(2) i.v. day 1), and prednisolone (30 mg orally days 1-7). RESULTS: The response rate was 52.2% including a complete response in four patients, and a partial response in eight patients. Adverse drug reactions included leukocytopenia (78.3%, 18/23 patients), alopecia (30.8%, 7/23), and peripheral neuropathy and generalized fatigue (26.1%, 6/23). In patients responding to the drug regimen, 50% survival was 29 months, and in those not responding it was 12 months. CONCLUSION: Combination treatment with mitoxantrone hydrochloride, vincristine sulfate and prednisolone is an effective treatment for breast cancer. PMID- 12764671 TI - A population model of epirubicin pharmacokinetics and application to dosage guidelines. AB - PURPOSE: To use a population approach to identify readily available clinical or biochemical characteristics that influence the pharmacokinetics of epirubicin and to develop new dosage guidelines based on these results. METHODS: Data were available from 109 patients with advanced breast cancer, 72 of whom were known to have liver metastases. They were treated with single-agent epirubicin 12.5 to 120 mg/m(2). Analysis was performed using the software package NONMEM and a three compartment model was fitted to the data. RESULTS: Individual clearance (CL) estimates ranged from 4 to 86 l/h and the final model included CL as a function of aspartate aminotransferase (AST): CL (l/h)=72.9-(72.9x0.135xlnAST). Inclusion of this factor reduced the interindividual variability in CL from 49% to 39%. Using a target AUC of 4000 ng.h/ml, the following doses were predicted to achieve this exposure with the greatest precision: AST <150 IU/l 125 mg; AST 150-250 IU/l 90 mg; AST 250-500 IU/l 60 mg; AST >500 IU/l 30 mg. These new guidelines were compared with three other guidelines based on serum bilirubin or AST concentrations and body surface area (BSA). The new guidelines achieved the target with greater precision (root mean squared error, rmse, 39.0%) than the current UK guidelines, current USA guidelines or an earlier equation based on AST (rmse 63%, 62% and 59%, respectively). CONCLUSIONS: The proposed dosing guidelines should reduce variability in systemic exposure to epirubicin more effectively than traditional approaches. In addition, as they do not require adjustment according to BSA, they could reduce dosage preparation time and the potential for prescribing and dispensing errors. PMID- 12764672 TI - Gemcitabine and vinorelbine as second-line treatment in patients with metastatic breast cancer: a phase II study. AB - BACKGROUND: To evaluate the feasibility and activity of gemcitabine and vinorelbine as a second/third-line approach in patients with advanced breast cancer. METHODS: Entered into the study were 51 consecutive patients. All had been previously treated with anthracyclines. Of these 51 patients, 36 had experienced failure or relapse after one chemotherapy line for advanced disease, and 15 after two chemotherapy lines. The dominant sites of involvement were brain in 4 patients (7.8%), liver in 22 (43.2%), lung in 10 (19.6%), bone in 10 (19.6), and soft-tissue in 5 (9.8%). Treatment consisted of vinorelbine 25 mg/m(2) and gemcitabine 1000 mg/m(2) administered on days 1 and 8 every 21 days. RESULTS: The scheme was well tolerated. Grade 3/4 neutropenia was observed in 11% of patients. Grade 3 nausea and vomiting occurred in 6%, and grade 2 neurotoxicity in 6%. No patients experienced grade 3/4 alopecia. The median relative dose intensity was 94.6% (49.7-100%) and 90.0% (23.1-100%) for vinorelbine and gemcitabine, respectively. Two patients (3.9%) were not evaluable for disease response, 4 (7.8%) attained a clinical complete response, 13 (25.5%) a partial response (for an overall response rate of 33.3%, 95% coefficient interval 20.0-46.0%), 23 (45.2%) showed stable disease, and 9 (17.6%) progressed. The median time to progression of responding patients was 10.8 months, and the median overall survival of the entire population was 17.8 months. CONCLUSIONS: Vinorelbine and gemcitabine is a manageable scheme with moderate activity in pretreated patients with advanced breast cancer. PMID- 12764676 TI - Hypertriglyceridemia associated with amino acid variation Asn985Tyr of the RP1 gene. AB - Factors predisposing to the phenotypic features of hypertriglyceridemia have not been clearly defined. Here we report an association between a missense coding region polymorphism Asn985Tyr in the retinitis pigmentosa 1 gene ( RP1) and plasma triglyceride (TG) levels in 332 adult individuals from an east-central area of Japan. Age and gender-adjusted levels of LDL-cholesterol, TG, and HDL cholesterol were analyzed. When we separate the subjects into two genotypic groups regarding this amino acid variation, those who lack the 985-Asn allele (asparagine at residue 985) had significantly higher plasma TG levels than the others who had at least one 985-Asn allele (mean: 175.8 mg/dl vs 123.3 mg/dl; P=0.0006, Mann-Whitney test). Similarly, the former subjects had significantly lower HDL-cholesterol levels than the latter (mean: 48.0 mg/dl vs 53.8 mg/dl; P=0.038). Of the 280 individuals without a 985-Asn allele, approximately half of the individuals presented with hypertriglyceridemia, whereas only a quarter were hypertriglyceridemic among 52 individuals with the 985-Asn allele ( P=0.04). Although this SNP marker may itself be in linkage disequilibrium with other unexamined functional variants within this locus, our data suggest that genetic variation at the RP1 locus is one of the likely candidate determinants for plasma triglyceride and HDL-cholesterol metabolisms. PMID- 12764678 TI - [Typical features of craniofacial growth of the FGFR3-associated coronal synostosis syndrome (so-called Muenke craniosynostosis)]. AB - BACKGROUND: The FGFR3-associated coronal synostosis syndrome (Muenke craniosynostosis) is caused by a point mutation (C749G) on the FGFR3 gene resulting in a Pro250Arg substitution. METHODS: To characterize this malformation, the neuro- and viscerocranium were analyzed by axial CT scans of the skull and cephalometric radiographs of up to 13 affected children before and in part after fronto-orbital advancement. RESULTS: Preoperative analysis of the intracranial volume of four patients showed a mean decrease of 3.6%, indicating a compensatory growth pattern of the skull in cases of coronal synostosis. The typical brachycephaly could be verified by the significant shortening of the skull length of 13.2% on average and by the significant reduction of the anterior cranial base length of maximal 5.9% on average. The anterior part of the skull was characterized by a significant mean increase of the intercoronal distance of 8.6%, which indicates a compensatory transversal growth in this malformation. The widened bilateral interorbital and anterior interorbital distances were increased by 7.3 or 9.0%, respectively, confirming a hypertelorism typical for this syndrome. The "frontal bossing" frequently found in brachycephaly was characterized by the preoperatively increased sagittal extension of the forehead (about 112.9% above the norm) and by the increased height of the frontal prominence (about 47.8% above the norm). Following surgery, both variables defining the morphology of the forehead were reduced and appeared to be constant throughout the follow-up. Hypoplasia of the midface described by Muenke et al. (1997) was confirmed in the present study only by the significant reduction of the sagittal length of the maxillary base, which was decreased by 6.8%. CONCLUSION: In accordance with the current literature, the skull configuration described for Muenke craniosynostosis shows similarities with the Saethre-Chotzen syndrome. PMID- 12764679 TI - [Evaluation of her-2/neu amplification/overexpression in OSCC with fluorescence in situ hybridization (FISH) and immunohistochemistry]. AB - INTRODUCTION: The human epidermal growth factor receptor ( her-2/ neu) protooncogene encodes a membrane tyrosine kinase with homology to the epidermal growth factor receptor (EGFR). Amplification and protein overexpression have been identified in various solid tumors and a significant association with poor clinical outcome was reported. This investigation was performed to assess the frequency of her-2/ neu overexpression and to compare these results with clinical outcome in OSCC. MATERIAL AND METHODS: Archival biopsy specimens from 97 untreated OSCCs were evaluated using a polyclonal antibody A0485 (Dako). Only membrane staining intensity and pattern were evaluated according to the guidelines of the clinical trial assay recommendations (0-3+) for breast carcinoma. Score 0 and 1+ were interpreted as negative for HER-2/NEU protein overexpression and 2+ and 3+ as positive. FISH analysis with directly labeled probes for her-2/ neu and chromosome 17 was performed on the same specimens. The ratio between her-2/ neu and chromosome 17 signals was calculated after selection of 20-40 non-overlapping tumor cells. The tumor was considered amplified if the ratio was above 2. RESULTS: In 11 out of 97 biopsies (11.3%) membranous overexpression (score 2+ and 3+) of her-2/ neu was shown by immunohistochemistry. FISH analysis in 42 cases revealed amplification in 14 cases. Concordance between immunohistochemistry and FISH was found in 86%. Clinical-pathological data as well as survival revealed no correlation with her-2/ neu status. DISCUSSION: In spite of missing correlation between survival and her-2/ neu overexpression in our study, the predictive value of the her-2/ neu protooncogene in adjuvant therapy in OSCC needs further investigation. PMID- 12764680 TI - [James Israel (1848-1926). His contribution to oral and maxillofacial surgery]. AB - The Berlin surgeon James Israel (1848-1926) is generally known as one of the leading kidney surgeons of his time. Furthermore, he is commemorated for describing actinomycosis bacteria, named after him in 1878: Actinomyces israelii. It is less well known that he also made several important contributions to maxillofacial surgery. This special aspect as well as Israel's biography is presented in this article from the history of medicine. PMID- 12764681 TI - [Memory effect of resorbable polymers]. AB - BACKGROUND: The present study analyzed the memory effect of resorbable polymers. Depending on temperature, this effect describes the ability of different materials to "remember" their original form after mechanical deformation. Resorbable polymers serve as materials to stabilize and fix bone fractures. Compared to metal transplants, resorbable polymers are able to undergo thermoplastic deformation. The precise adaptation of the transplant to the surrounding bone guarantees an exact anatomical reconstruction. However, during normal applications, it was observed that these biodegradable plastic materials tend to revert to their original form at body temperature. This "memory effect" could result in negative consequences for the medical treatment. METHODS: By the process of compression molding, geometrically formed specimens (lattice, rod, plate) consisting of different polyglycolides and polylactides were prepared. After warming up the specimens to 50 degrees C (water bath) they were deformed into definite angles. Following this procedure, the specimens were put in a water bath at 37 degrees C to mimic the adaptation of the transplant at body temperature. The retroflexion of the material (memory effect) was measured using an XY-measuring desk. RESULTS: The present study clearly reveals that highly deformed specimens react with stronger retroflexions. In addition, the results indicate that the memory effect depends on geometrical design as well as on chemical composition. All tested polymers showed a strong initial memory effect that decreased with time. PMID- 12764682 TI - [Comparative study of spatial imaging techniques in stereo-endoscopy]. AB - PURPOSE: This study compares the two-dimensional presentation of stereo endoscopic video data with three-dimensional presentation using polarization glasses and three-dimensional presentation with an autostereoscopic display. The aim of this study was to evaluate possible advantages of the three display technologies. MATERIAL AND METHODS: Fifty-nine test persons untrained in endoscopy had to complete three endoscopic tasks with different levels of difficulty. Each test involved a new presentation method. Different measurements were noted such as speed of task completion, accuracy of task performance, and quantity of solved tasks. The data collected were statistically evaluated. RESULTS: Neither sex, handedness, nor level of stereopsis had any statistically significant impact on the test results. The differences between the three presentation methods of stereo-endoscopic pictures were not statistically significant. CONCLUSION: Similar results were achieved with all three presentation methods. None of the presentation methods was significantly superior in the values measured. A final assessment of the possibilities of spatial endoscopy should await future technological developments in endoscopic devices (e.g., picture acquisition). PMID- 12764683 TI - [Proliferative verrucous leukoplakia. Report of five cases]. AB - BACKGROUND: Proliferative verrucous leukoplakia (PVL) is a rare variant of oral leukoplakia (OL) with a high tendency for malignant transformation. PVL starts as a flat hyperkeratosis. During the course of the disease verrucous lesions develop. AIM OF THE STUDY, MATERIAL AND METHODS: Due to the scarcity of reports on PVL five cases of PVL are presented. Clinical and histopathological findings in five female patients with PVL were evaluated retrospectively. RESULTS: The clinical appearance of PVL was characteristic as compared to criteria described in the literature. The average period of observation in these cases was short (1.7 years). There were 3.6 different localizations observed per patient. The buccal mucosa, gingiva, and edentulous alveolar ridge were most often affected. One patient had already developed squamous cell carcinoma prior to referral. The others developed malignancies during follow-up. Recurrences and secondary malignancies could not be avoided using different therapeutic modalities. Two patients died from PVL. The characteristic spectrum of histopathological findings in PVL was observed. Homogeneous flat leukoplakias are characterized by acanthosis, hyperkeratosis, and often parakeratosis. Epithelial dysplasia is not seen. The proliferative verrucous stage of the disease is characterized by papillomatous epithelial growths with signs of slight epithelial dysplasia and subepithelial infiltration with immunocompetent cells. Stages of transformation may result in verrucous carcinoma or squamous cell carcinoma. CONCLUSIONS: PVL is associated with a very high rate of malignant transformations. These, as well as recurrences, cannot be avoided with present-day therapies. In order to define the biological profile of PVL more clearly, multicenter studies are necessary to reveal possible etiologic factors and concepts for therapy. PMID- 12764674 TI - Methodologies for the characterization of microbes in industrial environments: a review. AB - There is growing interest in research and development to develop novel tools to study, detect, and characterize microbes and their communities in industrial environments. However, knowledge about their validity in practical industrial use is still scarce. This review describes the advantages and limitations of traditional and molecular methods used for biofilm and/or planktonic cell studies, especially those performed with Listeria monocytogenes, Bacillus cereus, and/or Clostridium perfringens. In addition, the review addresses the importance of isolating the microorganisms from the industrial environment and the possibilities and future prospects for exploiting the described methods in the industrial environment. PMID- 12764684 TI - [Inflammatory paradental cyst. Report of 6 cases]. AB - BACKGROUND: The inflammatory paradental cyst has been described as an entity in the WHO classification of odontogenic tumors and cysts (1992). It is mainly located at mandibular molars, in particular third molars of the lower jaw. Radiologically, involved molars show a circumscribed, mostly half-moon shaped translucency distal or distobuccal to the involved tooth. Patients frequently report episodes of infection (pericoronitis). The histological findings are identical to those of inflammatory radicular cysts. The inflammatory paradental cyst has been described infrequently in the international literature. There are no reports available in German. AIM OF THE STUDY AND CASES: The aim of the present study was to present six of our own cases of inflammatory paradental cysts. Five men and one woman with an average age of 29.5 years were affected. In two cases paradental cysts occurred bilaterally. Three patients reported recurrent previous infections (pericoronitis). Radiologically, the typical translucency with clear demarcation distal to the third molars was observed. All of the third molars were vertically retained. Histologically, the inflammatory paradental cysts showed features identical to those of radicular cysts. The inflammatory paradental cyst is a clear indication for osteotomy of lower wisdom teeth. Postoperative complications or recurrences of the inflammatory paradental cysts have not been described. DISCUSSION: A correct clinical, radiological, and histopathological diagnosis of paradental cysts is mandatory, and more reports are needed in order to compile more information about relative frequency and pathogenesis of this cyst variant. PMID- 12764685 TI - [Acute tongue necrosis provoked by epirubicin-cyclophosphamide treatment for invasive ductal breast cancer]. AB - CASE: Unilateral necrosis of the tongue is an uncommon symptom of different rare diseases. Previously, it had only been described as an infrequent complication of temporal arteritis or as a side effect of therapy with ergotamine. We present a case of unilateral necrosis of the tongue in a 62-year-old woman with invasive ductal carcinoma of the breast treated with epirubicin and cyclophosphamide. RESULTS: The necrosis led to a rapid swelling of the tongue and consequently to an airway obstruction necessitating a tracheotomy. After excision of the necrosis, the swelling of the tongue and the airway obstruction subsided. DISCUSSION: Because of the temporal connection between the occurrence of the necrosis and the administration of chemotherapy, an adverse effect of the administered drugs seems most likely. However, a paraneoplastic pathogenesis cannot be completely excluded. The occurrence of unilateral necrosis of the tongue is a rare complication of the above-mentioned conditions. However, it is important to be aware of the different causes leading to this rare disease in order to initiate the right therapy. PMID- 12764686 TI - Epithelioid hemangioendothelioma of the mandible. AB - The authors describe a locally recurrent tumor of the mandible whose microscopic structure and immunohistochemical findings corresponded to an epithelioid hemangioendothelioma. The tumorous cells had an epithelioid character, they created rudimentary vascular lumina, and they focally expressed vascular markers CD 34, CD 31, and F VIII. Locally the tumor grew aggressively. During the last recurrence, the tumor even affected the already healed and rebuilt bone graft, which was incorporated after the primary resection of the mandibular body. From the histological point of view, the tumor approximated an angiosarcoma. No metastases occurred. PMID- 12764687 TI - Molecular modeling of penicilloate anions: an RHF-SCF analysis. AB - An ab initio restricted Hartree-Fock self-consistent field (RHF-SCF) analysis of penicilloate anions was performed at the TZV level with GAMESS. Geometry optimization was initialized by the semi-empirical AM1 method followed by optimization at the 6-31++G** level. The total energy obtained was -1116.0997 a.u. for the penicilloate amine, -1115.3164 a.u. for the imine, -1115.2969 a.u. for the enamine and -1115.2017 a.u. for the amine that was deprotonated at the thiazolidine nitrogen. Formation of the free thiolate in the imine and enamine anions by deprotonation of the penicilloate amine is associated with: (1) an increase in total energy (2) an increase in the energy of the highest occupied molecular orbital (HOMO) to that of anti-bonding (3) a decrease in chemical hardness (4) an increase in the chemical potential (5) a more negative Mulliken net charge on the sulfur atom and (6) an increase in the Mulliken atomic population on the former thiazolidine sulfur atom in the HOMO. The RHF-SCF analysis presented here suggests a potential role for the thiolate sulfur of penicilloate anions, especially of the imine, as a chemically reactive soft nucleophile. PMID- 12764688 TI - Spectroscopic characterization of the [Fe(His)(4)(Cys)] site in 2Fe-superoxide reductase from Desulfovibrio vulgaris. AB - The electronic and vibrational properties of the [Fe(His)(4)(Cys)] site (Center II) responsible for catalysis of superoxide reduction in the two-iron superoxide reductase (2Fe-SOR) from Desulfovibrio vulgaris have been investigated using the combination of EPR, resonance Raman, UV/visible/near-IR absorption, CD, and VTMCD spectroscopies. Deconvolution of the spectral contributions of Center II from those of the [Fe(Cys)(4)] site (Center I) has been achieved by parallel investigations of the C13S variant, which does not contain Center I. The resonance Raman spectrum of ferric Center II has been assigned based on isotope shifts for (34)S and (15)N globally labeled proteins. As for the [Fe(His)(4)(Cys)] active site in 1Fe-SOR from Pyrococcus furiosus, the spectroscopic properties of ferric and ferrous Center II in D. vulgaris 2Fe-SOR are indicative of distorted octahedral and square-pyramidal coordination geometries, respectively. Differences in the properties of the ferric [Fe(His)(4)(Cys)] sites in 1Fe- and 2Fe-SORs are apparent in the rhombicity of the S=5/2 ground state ( E/ D=0.06 and 0.28 in 1Fe- and 2Fe-SORs, respectively), the energy of the CysS(-)(p(pi))-->Fe(3+)(d(pi)) CT transition (15150+/-150 cm( 1) and 15600+/-150 cm(-1) in 1Fe- and 2Fe-SORs, respectively) and in changes in the Fe-S stretching region of the resonance Raman spectrum indicative of a weaker Fe-S(Cys) bond in 2Fe-SORs. These differences are interpreted in terms of small structural perturbations in the Fe coordination sphere with changes in the Fe S(Cys) bond strength resulting from differences in the peptide N-H.S(Cys) hydrogen bonding within a tetrapeptide bidentate "chelate". Observation of the characteristic intervalence charge transfer transition of a cyano-bridged [Fe(III)-NC-Fe(II)(CN)(5)] unit in the near-IR VTMCD spectra of ferricyanide oxidized samples of both P. furiosus 1Fe-SOR and D. vulgaris 2Fe-SOR has confirmed the existence of novel ferrocyanide adducts of the ferric [Fe(His)(4)(Cys)] sites in both 1Fe- and 2Fe-SORs. PMID- 12764690 TI - ST-T integral and T-wave amplitude in detection of exercise-induced myocardial ischemia evaluated with body surface potential mapping. AB - Body surface potential mapping is superior to 12-lead electrocardiogram in detection of acute and old myocardial infarctions. We examined the capability of the ST-T integral and T wave to detect exercise-induced ischemia in body surface potential mapping. Body surface potential mapping with 123 channels was recorded in 70 subjects: 45 coronary artery disease (CAD) patients and 25 healthy controls during supine bicycle exercise testing. Of the patients, 18 had anterior, 14 posterior, and 13 inferior ischemia documented by coronary angiography and thallium scintigraphy. The ST-T isointegral area, as well as the positive and negative ST-T area, and the T-wave apex amplitude were determined. Discriminant index analysis was used to find the sites that optimally separated patient subgroups from other patients and controls. In the pooled CAD group, the optimal sites for detecting the decrease in ST-T isointegral, in the positive ST-T area and in the T-wave amplitude were over the left side (ST-T isointegral area: CAD 3.8 +/- 14 microVs and controls 24 +/- 14 microVs; T-wave amplitude: CAD 3 +/- 110 microV and controls 190 +/- 90 microV; P <.001, both). The area under the receiver operating characteristic curve for the decrease in ST-T isointegral, in the positive ST-T area, and in the T-wave amplitude and for the ST depression were 94%, 95%, 92%, and 93%, respectively. T wave performed especially well in patients with multivessel disease. In stepwise logistic regression analysis, using the presence of CAD as the dependent parameter, the decrease in the positive ST-T area and ST depression were the only parameters that entered the model. ST-T area and T-wave amplitude are sensitive and specific markers of transient myocardial ischemia. ST-T area contains information additional to ST depression and has thus independent discriminative value in ischemia detection. PMID- 12764691 TI - QT dispersion increases during intubation in patients with coronary artery disease. AB - Anesthetic drugs have been shown to increase QT interval, however data regarding their effects on QT dispersion (QTd) are scarce, especially in patients with coronary artery disease (CAD). We tested whether induction of Anesthesia with thiopental and etomidate would increase QTd in patients with CAD. Thirty American Society of Anesthesiologists (ASA) physical status I-II patients without CAD were randomly allocated to groups I (n = 15) and II (n = 15) and 30 ASA physical status III patients with CAD were randomly allocated to groups III (n = 15) and IV (n=15). Anesthesia was induced with thiopental 5-7 mg/kg IV in groups I and III and with etomidate 0.2-0.3 mg/kg IV in groups II and IV. Endotracheal intubation was facilitated with vecuronium bromide 0.1 mg/kg IV. Twelve-lead electrocardiogram (ECG) was recorded in all patients at baseline (ECG(1)), 1 min after the induction agent (ECG(2)), 1 min (ECG(3)) and 5 min (ECG(4)) after intubation. Anesthesia was maintained with isoflurane and nitrous oxide in 34 % oxygen after ECG(2) recording. QTd and corrected QT dispersion (QTcd) were calculated. In patients with CAD induced with thiopental, QT dispersion increased significantly during the intubation period compared with baseline (from 43.0 +/- 25.6 ms to 69.2 +/- 25.3 ms; P <.01). Likewise, QT dispersion also increased during intubation in patients with CAD induced with etomidate (from 41.5 +/- 17.2 ms to 80.0 +/- 33.6 ms; P <.001). There was no increase in QT dispersion in patients without known CAD. QT dispersion seems to be increased during the intubation period in patients with CAD regardless of the induction agents used. PMID- 12764692 TI - Localization and radiofrequency ablation of atriofascicular pathways using electroanatomic mapping. AB - Atriofascicular pathways supporting antidromic reentrant tachycardia are uncommon, and may be difficult to ablate. Traditional mapping can be associated with traumatic loss of atriofascicular conduction. Atriofascicular fibers can insert into the right bundle and will, therefore, first activate the right ventricle. In contrast to initial activation of the ventricle near the tricuspid annulus that can be seen in patients with right-sided decremental atrioventricular pathways. We used electroanatomic mapping to map and ablate the ventricular insertion of atriofascicular pathways in two patients during sinus rhythm and during atrial pacing. In our 2 cases an atriofascicular potential was recorded from below the tricuspid valve annulus and tagged. At this site, each pathway was ablated with one radiofrequency lesion. We describe 2 cases where electroanatomic mapping of the right ventricle was used to map and ablate atriofascicular pathways. PMID- 12764693 TI - Long-term effects of beta blocker therapy on P-wave duration and dispersion in congestive heart failure patients: a new effect? AB - Atrial fibrillation (AF) is associated with heart failure in approximately 20% 50% of patients with increased morbidity and mortality. P-wave maximum duration (PWM) and P-wave dispersion (PWD) are recent ECG markers and reflect increased risk of AF. The aim of our study was to investigate the long-term effects of metoprolol on atrial conduction abnormalities as estimated by PWM and PWD, which were calculated on 12-derivation surface ECG in heart failure patients. Forty-two NYHA class 3 to 4 patients (23 males, 19 females and aged 52.9 +/- 11.2) were enrolled in the study. At the end of the 6 months, PWM and PWD values were significantly decreased (for PWM: from 115.2 +/- 12.6 to 105.4 +/- 13.5; for PWD: from 39.3 +/- 9.1 to 28.6 +/- 10.5; P <.001 for both). Metoprolol treatment is associated with a decreased duration of PWM and PWD and this may reflect a reduction in the probability of atrial fibrillation in heart failure patients. PMID- 12764694 TI - Wavelet transform analysis of heart rate variability to assess the autonomic changes associated with spontaneous coronary spasm of variant angina. AB - We used Wavelet transform (WT) to investigate whether variation in autonomic tone was associated with spontaneous coronary spasm in patients with variant angina by analysis of heart rate variability (HRV). Twenty-one episodes preceding ST segment elevation were selected under Holter monitoring in 12 men and 3 women with variant angina. HRV indices were calculated at 10 second intervals with the continuous WT, and analyzed within 30 minutes preceding ST-segment elevation. High frequency (HF; 0.15 approximately 2.00 Hz) increased significantly during the 4 minutes prior to ST-segment elevation, low frequency (LF; 0.04 approximately 0.15 Hz) decreased significantly during the period from 10 to 5 minutes and increased significantly during the 2 minutes prior to ST-segment elevation, the LF/HF ratio decreased significantly during the period from 10 to 3 minutes and increased significantly during the 2 minutes prior to ST-segment elevation. The RR interval decreased significantly during the 2 minutes prior to ST-segment elevation. These results suggest that the acute variation in autonomic tone was associated with spontaneous coronary spasm in patients with variant angina. A reduction in sympathetic activity, then enhancement of vagal activity may play a key role in triggering the spontaneous coronary spasm, and the secondary activation of sympathetic activity may worsen the coronary spasm resulting in the attack. PMID- 12764695 TI - Possible contribution of the sarcoplasmic reticulum Ca(2+) pump function to electrical and mechanical alternans. AB - We investigated the role of the sarcoplasmic reticulum's (SR) Ca(2+) pump function of the in the mechanism of alternans. We recorded the surface ECG, monophasic action potential (MAP) and left ventricular pressure (LVP) in the canine beating heart. Alternans was induced with an abrupt shortening of the cycle length from 1000 to 350 ms. After the control studies, we administered propranolol or isoproterenol. In the presence of propranolol, we administered milrinone or 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS). In the presence of isoproterenol, we administered thapsigargin. Isoproterenol and milrinone attenuated both the electrical and mechanical alternans. Thapsigargin, a specific SR Ca(2+) pump inhibitor, and propranolol magnified both types of alternans. DIDS, a Ca(2+)-activated Cl(-) current (I(Cl(Ca))) inhibitor, attenuated the MAP alternans without an affect on the LVP alternans. Thus, the delayed intracellular Ca(2+) cycling caused by the impaired SR Ca(2+) pump function might produce electrical and mechanical alternans. beta-adrenergic stimulation eliminated these alternans. The I(Cl(Ca)) contributed to the appearance of the electrical alternans. PMID- 12764696 TI - Increased cycle length variability during ventricular fibrillation: a novel predictor of arrhythmia recurrence. AB - To evaluate the clinical value of cycle length (CL) variability during ventricular fibrillation (VF), 26 patients who underwent implantable cardioverter defibrillator (ICD) implantation were enrolled. In VF induced for defibrillation testing, mean and SD of VFCL, mean successive differences (MSD) of VFCL, and coefficient of variations of the VFCL (CV(FF)) (SD x 100/mean VFCL) were calculated. During the follow-up period of 20 +/- 2 months, ventricular arrhythmias recurred in 13 patients. MSD and CV(FF) were 31 +/- 3(*) ms and 15.6 +/- 1.3(**) in recurrence group (n = 13), and 17 +/- 2 ms and 9.0 +/-1.1 in non recurrence group (n = 13) ((*)P <.005, (**)P <.001 vs. nonrecurrence group). Relatively good repeatability of mean VFCL, MSD and CV(FF) in each patient was confirmed by the Bland-Altman method. In VF induced by programmed ventricular stimulation before ICD implantation, MSD and CV(FF) in recurrence group were also increased significantly. Kaplan-Meier estimates revealed that MSD >or= 20 ms and CV(FF) >or= 12 predicted higher arrhythmia recurrence (MSD, P =.039; CV(FF), P =.0069 by the log-rank test). By multivariate analysis, CV(FF) >or= 12 was a significant predictor of recurrent arrhythmic events (P =.019). In conclusion, the CL variability of VF, which was evaluated as MSD and CV(FF), is increased in patients with arrhythmia recurrence. These values may reflect the degree of electrical heterogeneity, and appears to be useful indexes of the future arrhythmic events. PMID- 12764697 TI - Two-dimensional analysis of ventricular fibrillation in the guinea pig. AB - Cardiac arrhythmias are undesirable electrical activity in the heart. Ventricular fibrillation (VF) is a fatal cardiac arrhythmia and is characterized by the breakdown of organized electrical activity in the ventricular myocardium. However, little is known about VF, partially because it is difficult to study and understand an apparently disorganized activity. One method for discovering the nature of VF is processing the epicardial electrical signals by using cardiac mapping techniques. These techniques involve the study of propagation patterns seen in VF. In this project, we used guinea pigs to study the spatial organization of the epicardial electrical activity during VF. VF was induced in 9 open chest guinea pigs and epicardial electrode data were acquired by using a square array of 192 electrodes for 4 seconds every minute. The mean correlation length and dominant frequency were measured in each recorded segment. Correlation length is a measure of the spatial order in a system and dominant frequency is the frequency corresponding to the spectral maximum. The mean correlation length was found to vary between 1-7 mm and the dominant frequency was in the range of 1 14 Hz. This study suggests that guinea pig VF exhibits a level of organization equivalent to or greater than that seen in previous studies. PMID- 12764698 TI - Electrocardiographic findings in naturally acquired chagasic heart disease in nonhuman primates. AB - The significance of electrocardiographic (ECG) changes described in animals with Chagas' disease is questionable in view that other non-invasive comparisons have been lacking. 12-lead ECG and two-dimensional echocardiography (echo) was performed in 17 seropositive and 13 seronegative baboons. Similar to humans, a wide variety of ECG outcomes were observed in the infected animals. Standard ECG measurements were not different between groups. Five seropositive (29%) and 3 seronegative (23%) animals had low voltage; 4 seropositives (24%) and 2 (15%) seronegatives had tall P-waves. Precordial Q waves were seen in 10 seropositives (59%) and in 7 (54%) seronegatives without septal abnormalities on two dimensional echo. One seropositive animal had a 2(nd) degree (Wenckebach) AV block and left anterior fascicular block. Most animals in both groups had diffuse T-wave abnormalities. Echo evidence of systolic dysfunction was found in 4 seropositives and in none of the controls; thus, chagasic heart disease was present in 24% of naturally infected baboons. Since most non-human primates, irrespective of their serology, have diffuse, nonspecific ECG changes not necessarily diagnostic of myocardial disease, two-dimensional echo should be added to their non-invasive assessment. PMID- 12764699 TI - Cycle length-dependent repolarization changes during atrial fibrillation in the Brugada syndrome. AB - This is a case report of a patient with Brugada syndrome who developed paroxysmal atrial fibrillation. During the episode, beat-to-beat changes in ventricular repolarization were observed. These changes were a paradoxical ST-segment alteration after a short-coupled ventricular beat. These findings, not reported before, may be helpful for the diagnosis of this syndrome. PMID- 12764701 TI - Relationship among electrocardiographic potential amplitude, weight, and resistance/reactance/impedance in a patient with peripheral edema treated for congestive heart failure. AB - We are presenting a 79-year-old woman with congestive heart failure (CHF) and marked peripheral edema, who responded to medical management by losing 19.7% of her weight. The patient was studied with daily weights, 12-lead electrocardiograms (ECGs), and body electrical resistance/reactance/impedance (R/R/I) measurements via electrodes applied on her hand and foot. In the process she showed a gradual increase in the QRS complexes in her daily ECGs pari passu with the weight loss, leading to a 27.4% increase in the sums of the QRS complexes from the 12 ECG leads ( summation operator SQRS). In addition the patient revealed excellent correlations between weights and summation operator SQRS (r = -.91, P =.013), weights and resistance (r = -.95, P =.004), reactance (r = -.86, P =.029), and impedance (r = -.95, P =.004). Other ECG sets including fewer leads were also employed and correlated with weights and the body electrical properties. It is concluded that loss of peripheral edema in patients with CHF is associated with augmentation of body R/R/I leading to augmented ECG QRS complexes. These findings link etiologically changes of ECG potentials to alteration of body electrical properties, and suggest that serial ECGs could be used in the monitoring of therapy of patients with CHF. PMID- 12764702 TI - Electrocardiographic manifestations of proximal left anterior descending artery occlusion. AB - We report the case of a 51-year-old woman who presents with a 2-week history of episodes of pressure like chest pain. The initial electrocardiogram was not indicative of myocardial ischemia or infarction and the cardiac enzymes remained normal during the initial hospital day. However, the precordial T waves inverted and progressively deepened on the second hospital day and the patient underwent cardiac catheterization with percutaneous coronary angioplasty and stent placement of the left anterior descending coronary artery with good results. The postprocedure electrocardiogram showed complete resolution of the inverted precordial T waves. The development of new T-wave inversions in the precordial leads of patients presenting with unstable angina is predictive of significant stenosis of the left anterior descending coronary artery. This subgroup of patients has a poor prognosis if medical therapy alone is instituted. Early cardiac catheterization and revascularization is recommended for these patients. Evidence has shown that 75% patients with these electrocardiogram changes who are not revascularized developed extensive anterior wall infarction within a few weeks. PMID- 12764704 TI - Social and health functioning in female primary care patients with post-traumatic stress disorder with and without comorbid substance abuse. AB - The present study examined whether post-traumatic stress disorder (PTSD) and comorbid substance use disorder (SUD) is associated with greater social and health morbidity than PTSD without SUD in a sample of female primary care patients. Participants were administered diagnostic interviews and assessed for work productivity, quality of interpersonal relationships, and degree of health functioning. No significant differences were found between the women with current PTSD and a comorbid lifetime substance use disorder (N = 56) and those with current PTSD and no lifetime substance use disorders (N = 60) in degree of work productivity, interpersonal functioning, and overall well-being and health, as well as number of lifetime medical illnesses. These findings suggest that the presence of comorbid SUD may not explain the level of social and health difficulties associated with the dual diagnosis of PTSD and SUD. PMID- 12764703 TI - On the relation between neuroticism, self-esteem, and depression: results from the National Comorbidity Survey. AB - There is evidence that neuroticism and self-esteem, two commonly used personality constructs, are associated with depressive illness. Previous studies on this issue have produced mixed results. Some studies found that neuroticism was a stronger predictor of risk for major depression than was self-esteem. The aim of the current report is to analyze the relationship between neuroticism, self esteem, and depressive disorders in representative community sample. Data from the National Comorbidity Survey (NCS) were reanalyzed. The diagnostic data were coded using the criteria of DSM-III-R based on a modified version of the Diagnostic Interview. Self-esteem was assessed by an empirically abbreviated form of the Rosenberg Self-Esteem Scale. Neuroticism was measured using the 10-item scale from the Transparent Bipolar Inventory (TBI). Logistic regression analysis and classification and regression tree (CART) analysis were used to determine the associations between neuroticism, self-esteem, sociodemographic variables, and past-year depression disorders. Neuroticism and self-esteem were strongly associated with past-year depression disorders. Significant interactions between the two personality constructs and sociodemographic variables were observed. The results suggest that neuroticism and self-esteem should be evaluated simultaneously when analyzing depression disorders. The assessment of both personality constructs may contribute to further understanding of personality depression correlation. Such knowledge might prove valuable in designing early interventions and treatment. PMID- 12764705 TI - Externalizing disorders in consecutively referred children and adolescents with bipolar disorder. AB - We describe a consecutive clinical sample of children and adolescents with bipolar disorder (BD), in order to define the pattern of comorbid externalizing disorders and to explore the possible influence of such a comorbidity on their cross-sectional and longitudinal clinical characteristics. The sample consisted of 59 bipolar patients: 35 males and 24 females, with a mean age 14.6 +/- 3 years (range, 7 to 18 years), diagnosed as either type I or II according to DSM-IV. All patients were screened for psychiatric disorders using historical information and a clinical interview, the Diagnostic Interview for Children and Adolescents Revised (DICA-R). Severity and subsequent outcome of the symptomatology were recorded with the Clinical Global Impression (CGI), Severity and Improvement Scales, at the baseline and thereafter monthly for a period up to 48 months. BD disorder type I was present in 37 (62.7%) of the patients; 14 (23.7%) were affected by attention deficit-hyperactivity disorder (ADHD) and 10 (16.9%) by conduct disorder (CD). Comorbid ADHD was associated with an earlier onset of BD, while CD was highly associated with BD type I. Anxiety disorders appeared more represented in patients without CD. At the end of the observation, a lower clinical improvement was recorded in patients with CD. In our children and adolescents with BD, comorbidity with externalizing disorders such as ADHD and CD is common. The clinical implications of comorbid ADHD and CD are rather different. ADHD can be viewed as a precursor of a child-onset subtype of BD, while CD might represent a prodromal or a concomitant behavioral complication that identifies a more malignant and refractory form of BD. PMID- 12764706 TI - Coping strategies of panic and control subjects undergoing lactate infusion during magnetic resonance imaging confinement. AB - The psychological reactions and coping strategies used in response to a behavioral contingency (confinement in a magnetic resonance imaging [MRI] scanner) during a biological challenge (sodium lactate infusion) were systematically studied in 13 subjects with panic disorder (PD) and 11 control subjects using the Claustrophobia Questionnaire (CQ) and the Revised Ways of Coping Checklist (RWCCL). All participants were able to successfully complete the experimental procedure. Findings suggest between-group coping strategy differences in response to general stressors, but relative convergence of coping strategies in response to the experimental procedure, with relatively greater emphasis on problem-focused coping approaches. These observations suggest that PD patients are able to engage in effective coping strategies in response to stressful but highly structured experimental situations. An overall pre-post MR scanning reduction in the fear of restriction but not suffocation was observed for the combined sample, primarily reflecting changes in the control group. Among PD subjects, higher levels of suffocation fears were maintained despite emergence of more problem-focused coping in the experimental situation. PMID- 12764707 TI - Hispanic ethnicity, physician-patient communication, and antidepressant adherence. AB - The purpose of the study was to examine how Hispanic ethnicity influenced physician-patient communication about antidepressants and antidepressant adherence using a data set of audiotapes and transcripts of 98 medical visits and medical and pharmacy records. The data were collected in 1995 at the University of New Mexico's general medicine and family practice clinics. Physicians were more likely to state antidepressant information to non-Hispanic white patients than to Hispanic patients. Physicians were more likely to state information to patients who were prescribed new antidepressants. Physicians asked approximately one of five patients on continued therapy how well their antidepressants were working and only one of 10 patients if they were experiencing any side effects. Non-Hispanic white patients were significantly more likely to state information about their antidepressants than Hispanic patients. Younger patients and patients who were prescribed new antidepressants were more likely to ask questions about antidepressants. Hispanic patients and patients who were prescribed new antidepressants were significantly less adherent to their antidepressant therapy during the one hundred day period after their audio-taped visits than non Hispanic white patients and patients on continued therapy. PMID- 12764708 TI - Sexual coercion and abuse among women with a severe mental illness in India: an exploratory investigation. AB - Research from the west indicates that women living with a psychiatric disorder are particularly vulnerable to sexual coercion and abuse. However, there have been no published reports of sexual abuse among female psychiatric patients in India. This exploratory study sought (1) to determine the prevalence of sexual coercion in a representative sample of female psychiatric patients in India; (2) to identify clinical and sociodemographic correlates of sexual coercion; (3) to clarify the association between sexual coercion and human immunodeficiency virus (HIV)-related risk behavior; and (4) to determine whether self-report of sexual coercion from these patients was recorded in their medical charts. Consecutive female inpatient admissions (N = 146) to a large psychiatric hospital in southern India were assessed using a structured interview and standardized measures. During these structured clinical interviews, sexual coercion was reported by 30% of the 146 women. The most commonly reported experience was sexual intercourse involving threatened or actual physical force (reported by 14% of women), and the most commonly identified perpetrator was the woman's husband or intimate partner (15%), or a person in a position of authority in their community (10%). Women with a history of abuse were more likely to report HIV-related sexual behavior (P <.001). In contrast to the 30% of women who reported sexual coercion during interviews, only 3.5% of the medical records contained this information. Thus, sexual coercion is a serious and prevalent concern among female Indian psychiatric patients, but is rarely reported in medical charts. Increased screening and reporting are indicated, as are sexual abuse prevention and treatment programs. PMID- 12764709 TI - Interactive computer-training as a therapeutic tool in Alzheimer's disease. AB - The current study sought to evaluate a novel kind of interactive computer-based cognitive training (ICT) in Alzheimer's disease (AD). AD patients (N = 9), age- and gender-matched patients with a major depressive episode (N = 9), and healthy control subjects (N = 10) were trained to use an ICT program that relates to activities of daily living (ADL). Digital photographs of a shopping route were implemented in a close-to-reality simulation on a computer touch-screen. The task was to find a predefined shopping route, to buy three items, and to answer correctly 10 multiple-choice questions addressing knowledge related to the virtual tasks. Training performance was rated using the number of mistakes (wrong way), time needed for the tasks, number of correct multiple-choice answers, and of repeat of instruction. Compared to normal controls and depressed patients, AD patients performed significantly worse with regard to all variables. Within a 4 week training period including 12 sessions, however, substantial training gains were observed, including a significant reduction of mistakes. Training effects were sustained until follow-up 3 weeks later. The performance of the depressed patients and the normal controls improved as well, with no difference between the two groups. Self-reported effects revealed that the training was well perceived. Thus, the task performance of AD patients improved substantially and subjects appeared to have liked this approach to ICT. New interactive media, therefore, may yield interesting opportunities for rehabilitation and (psycho)therapeutic interventions. PMID- 12764710 TI - The brief symptom inventory and the positive and negative syndrome scale: discriminate validity between a self-reported and observational measure of psychopathology. AB - Concern within the literature has emerged from time to time arguing the poor validity of self-reported measures in psychopathology, namely, the reporting of psychotic experience. Although it is commonly believed that patients who have had a psychotic episode cannot accurately self-report their experience, very few studies have been conducted to measure the concordance between self-reported and observational measures of psychopathology using multivariate statistical techniques. Sixty-nine patients presenting their first psychotic episode were interviewed and assessed on the Positive and Negative Syndrome Scale (PANSS) and were asked to complete the Brief Symptom Inventory (BSI). By clustering symptom dimensions from the BSI into discriminate functions, the research demonstrated that these symptom dimensions could adequately classify high versus low scores on the PANSS subscales and total score. When the same clusters were entered into multivariate analysis of variance (MANOVA) models, they also demonstrated significant differences between high versus low observed symptomatology on the PANSS Positive and General Subscale Groups and Total Score Groups. The current findings shed some doubt on the supposition that those who experience psychosis are unable to report symptom dimensions that concord with those who observe the psychosis. It appears that models, operational definitions, and the language used in measuring psychopathology may differ significantly from those who experience the psychotic experience and those who observe it. Techniques such as multitrait multimethod are discussed as ways of overcoming these concerns. PMID- 12764711 TI - Do psychiatrists understand research-related experiences, attitudes, and motivations of schizophrenia study participants? AB - Psychiatrists serve in different but important capacities with research participants. Because ethical problems may arise in research when participants misunderstand motives and responsibilities of clinical-investigators (i.e., "therapeutic misconception"), it is important that relationships between research participants and psychiatric caregivers and investigators be based on genuine mutual understanding. Considerations about attunement of psychiatrists with schizophrenia research participants have never been studied empirically. As part of a larger structured interview-survey study, we assessed the predictions of psychiatrists regarding self-reported experiences, attitudes, and motivations of individual schizophrenia research volunteers who were known to the respondent psychiatrists. Fourteen patient-psychiatrist pairs were identified for inclusion. Psychiatrists predicted schizophrenia research participants' ease of participation, privacy concerns, and willingness to participate in future research but underestimated their positive overall experience. Psychiatrists also were attuned to schizophrenia participants' attitudes and motivations regarding personal benefit but did not accurately predict their responses concerning helping others and science. Also, psychiatrists underestimated the schizophrenia participants' perceived degree of influence by researchers and caregivers during recruitment. Psychiatrists recognized the burden, primary concerns, future participation willingness, and personal benefit orientation of schizophrenia participants, but they did not appear to fully understand the overall positive quality of research experience, altruistic orientation, and relationship orientation of their schizophrenia patients who volunteer for research participation. PMID- 12764712 TI - Endocrine factors in the etiology of postpartum depression. AB - This article reviews the literature regarding endocrine factors postulated or presumed to be relevant in postpartum depression (PPD), a condition affecting at least 10% of childbearing women. The phenomenology and epidemiology of PPD are also described. Data suggest that parturition-related endocrine changes are causally implicated in PPD in a vulnerable subgroup of women. More specifically, studies by our group and others suggest a role for changes in estradiol and progesterone in precipitating mood symptoms among women with PPD. The mechanisms underlying such differential sensitivities remain undetermined. Future directions for research are explored. PMID- 12764713 TI - Cross-validation, predictive validity, and time course of the Benzodiazepine Dependence Self-Report Questionnaire in a benzodiazepine discontinuation trial. AB - The Benzodiazepine Dependence Self-Report Questionnaire (Bendep-SRQ) measures the severity of benzodiazepine (BZ) dependence on four domains: awareness of problematic use, preoccupation with the availability of BZ, lack of compliance with the therapeutic regimen, and withdrawal. Although promising results of the Bendep-SRQ have been obtained in cross-sectional studies, no attention has been paid to its clinical relevance during BZ withdrawal, i.e., predictive validity and time course. We performed cross-validation and evaluated the predictive validity and time course on 180 long-term BZ users who were taking part in a general practice BZ discontinuation trial. Three of the four domains had good scalability. Some concerns arose about the preoccupation scale, which emphasizes the need for cross-validation in clinically relevant populations. All scales showed excellent reliability (subject discriminability, item discriminability), while construct and discriminant validity were adequate. All four scales contributed significantly to the prediction of whether complete abstinence would be achieved directly after taking part in the discontinuation program. This prediction was independent of the other prognostic variables, except for those in the domain problematic use. The scales problematic use and preoccupation showed good sensitivity to changes during follow-up. The insensitivity of the scale, lack of compliance can be explained by low baseline scores in our population, while the insensitivity of the withdrawal scale was probably the result of the study design. In conclusion, our study indicated the clinical relevance of the Bendep-SRQ before and during a BZ discontinuation trial. We recommend the use of the Bendep-SRQ in discontinuation therapy and research into the field of BZ addiction. PMID- 12764714 TI - The relation of psychogenic excoriation with psychiatric disorders: a comparative study. AB - Psychogenic excoriation (PE), characterized by excessive scratching or picking of the skin, is not yet recognized as a symptom of a distinct DSM-IV disorder. It is a chronic disorder with a high rate of psychiatric comorbidity. The purpose of this study was to compare patients diagnosed with PE and patients with another dermatological disease in terms of comorbid psychiatric disorders. Thirty-one consecutive subjects were recruited from an outpatient dermatology clinic. The control group was composed of 31 patients with chronic urticaria. All subjects were interviewed using the Structured Clinical Interview for DSM-III-R (SCID-I), Beck Depression Inventory (BDI), Hamilton Anxiety Rating Scale (HARS), and Yale Brown Obsession and Compulsion Scale (Y-BOCS) and also completed a semistructured questionnaire. Current major depressive syndrome was the most common psychiatric disorder in the PE group. There was a statistically significant difference between the two groups in terms of current major depressive syndrome (PE group 58.1%, control group 6.5%, P<.01). In the PE group, 45.2% of subjects were diagnosed with obsessive compulsive disorder (OCD), while the rate of OCD was only 3.7% in the control group (P <.01). The PE group scored significantly higher on the BDI, HARS, and Y-BOCS. The results of this study point to the close relationship of PE to depression and OCD. PMID- 12764717 TI - Connections among authors, readers, and editors...learning from mistakes. PMID- 12764718 TI - Elderly patients with a hip fracture: the risk for delirium. AB - This prospective study investigated risk factors for delirium in elderly hip fracture patients that could be recognized by nurses. Data were collected on predisposing and precipitating factors for delirium from 92 elderly patients with a hip fracture. Predisposing factors included age, gender, sensory impairments, functional impairment before the hip fracture, residency before admission, pre existing cognitive impairment, comorbidities, and medication use. Precipitating factors included factors related to surgery and to the postoperative period. Factors related to surgery included time between admission and surgery, type of surgery, type of anesthesia, duration of surgery and anesthesia, and complications during surgery. Factors studied in the postoperative period were slow recovery, malnutrition, dehydration, addition of three or more medications, introduction of bladder catheter, infections, complications and falls, and use of morphine. Eighteen patients developed delirium, as diagnosed by a geriatrician by using the Diagnostic Statistical Manual-IV criteria. Data on delirious patients were compared with the data on non-delirious patients. The findings confirm that elderly hip fracture patients with premorbid ADL dependency, psychiatric comorbidities (including dementia), and a high number of other comorbid problems are at risk for the development of delirium. Based on these findings, it is recommended that nurses should assess patients' pre-fracture functional and cognitive capacities in an early stage of the hospital stay. Nurses should also be alert to postoperative delirium in "healthy elderly" patients. Monitoring of symptoms postoperatively in all elderly patients is advised. PMID- 12764719 TI - ICU nurses' preparation of families for death of patients following withdrawal of ventilator support. AB - Intensive care unit nurses were asked how they prepared families for the death of their patient following withdrawal of mechanical ventilation. Forty-three descriptors were identified, of which 67.5% (n = 29) were "physical sensations and symptoms." Less frequently mentioned features of Self-Regulation Theory were temporal characteristics, environmental features, and causes of these signs. Eight descriptors mentioned by more than 50% of nurses were skin color changes (74%), skin temperature changes (74%), varying levels of consciousness (74%), effort with breathing (71%), variable timeframe to death (68%), breathing pattern (65%), sound during breathing (61%), and loss of bowel control/incontinence (52%). PMID- 12764720 TI - Beliefs about tobacco among Appalachian current and former users. AB - Higher risks of tobacco-related morbidity and mortality in Appalachians provided the impetus for this qualitative study. The purpose was to describe beliefs about tobacco use and cessation among current and former tobacco users in rural Appalachia using focus groups. Findings included themes of nicotine addiction and pros and cons of tobacco use. The importance of family and personal independence in relation to tobacco were evident. Participants identified helpful tobacco treatment strategies. Similarities to treatment needs of tobacco users in general underscore the point that rural Appalachians require best practice tobacco treatment integrating unique cultural aspects of family and personal independence. PMID- 12764721 TI - Education and support needs of younger and older cancer survivors. AB - This study examined the use, satisfaction with, and need for cancer education and support services in northeastern Pennsylvania. Responses of 243 younger survivors (40 to 64 years) were compared with 295 older survivors (> or = 65 years). Programs attended and desired were related to cancer type and age. The almost 20% of respondents who had attended programs, primarily younger survivors, expressed high levels of satisfaction. Younger survivors reported non-attendance because of lack of interest. Older survivors' non-attendance reasons included difficulty with travel (N = 259, chi(2) = 12.081, p =.002), no identified need for information, and dislike for time of offerings (evenings). Education/support programs should be targeted to the age of people being served, as well as incidence, prevalence, expected trajectory, and prognosis of cancer types in the service area. PMID- 12764722 TI - Burden of family caregivers with schizophrenic patients in Korea. AB - The purpose of the study was to test a staged causal model as the theoretical base to determine the relationships among knowledge, coping, and burden among Korean family caregivers with schizophrenic patients. The staged theoretical model contained three stages comprised of contextual variables (stage 1), interactional variables (stage 2), and perception variables (stage 3). The situational variables were caregiver knowledge, gender and age of family caregiver, duration of family caregiving, and the nature of the relationship between patient and family caregiver. The interactional variable was represented by two styles of copings (positive and negative). The perception variable was the perception of subjective burden. A total of 57 family caregivers with schizophrenic patients participated in this study. The instruments, Knowledge Scale, Coping Scale, and Burden Scale, were used. A path analysis was used in this model. The family caregiver's knowledge had an indirect impact on the burden through negative coping, indicating that the less caregiver's knowledge, the more caregivers use negative coping strategies, which results in caregiving perception of subjective burden. The results support that interactional outcome of coping mediates the relationships between caregiver's knowledge and the impact of subjective outcome of caregiving burden. PMID- 12764724 TI - Quality of life for caregivers and stroke survivors in the immediate discharge period. AB - Caregiver burnout leads to a decline in caregivers' quality of life and in the quality of care they can deliver. A sample of 22 stroke survivors and informal care givers from a 500-bed metropolitan teaching hospital were interviewed using a questionnaire. Pre- (at discharge) and post- (1 month later) test scores were tested using Wilcoxon rank tests. Associations between activities of daily living and quality of life were tested with Spearman's correlation coefficient. Over the month, caregivers' scores for relationships, social companionship, and daily emotional support decreased (but not significantly). Short hospital stays affected the recruitment for this study and contributed to problems for ward staff in implementing discharge plans. More innovative discharge planning is needed if caregivers' needs are to be met. PMID- 12764723 TI - Recruiting and screening older, transitional to frail adults in congregate living facilities. AB - Historically older adults have been recruited from the community or from long term care facilities. The research study, The Effects of Intense Tai Chi Training in the Older Adult, targeted adults 70 years and older who were residing in congregate living facilities and transitioning to frailty according to the Speechley/Tinetti scale. This article describes the strategies that were designed and implemented to recruit and screen the 287 older adults who entered the randomized, single-blinded clinical trial. PMID- 12764725 TI - Measuring nursing services in patient safety research. PMID- 12764726 TI - [The integrated approach between Health Organisation accreditation and professional excellence accreditation]. AB - Later than in other industrialized areas, Health Organisation accreditation has become popular in our country during the last few years as a result of legislation requiring accreditation for healthcare providers of the National Health Service. Out of the two accreditation models, for regulation and public accountability (institutional) and professional, voluntary self-developed (for excellence), Institutional accreditation process has been slowly growing with the adoption of generic structural and organisational standards, which have been set up and managed directly by government. The recently undertaken second model is a peer review assessment activity that originates from a comparison with explicit standards during reciprocal visits carried out by professional organisations. The basis for the assessment is primarily clinical and patient oriented. The healthcare quality of Institutional accreditation is guaranteed mainly through the achievement of predetermined standards for risk management and organisation. On the other hand, the excellence accreditation, whose aim is to implement the continuous improvement of quality, depends on the context and the changes that result from new knowledge and scientific progress. The model operates according to values, which can change in the long run. It measures the effort to improve performances while special care is required to define standards through peer consensus. Once the prescribed model of accreditation is adopted, as is the case for Health Organisation accreditation, it is essential to have flexible patient oriented tools ensuring that new knowledge and scientific progress complete the assessment and audit. PMID- 12764727 TI - [The meaning and implementation of clinical governance]. AB - Under the 1999 Health Act a statutory duty of "quality" was given to National Health Service (NHS) organisations in the United Kingdom. This was matched by a comprehensive quality program. In this paper we look at the meaning of clinical governance as a mechanism for ensuring local delivery of high quality clinical care in the UK, the national structures which have been put in place to develop, reinforce and implement clinical governance and the role of the NHS Clinical Governance Support Team (CGST) in delivering clinical governance "on the ground". As part of the quality program, the CGST is working to enable national clinical governance policy to be translated into practice locally by supporting the development of better ways of working by individuals, clinical teams and health organisations to deliver a continuous, integrated approach to quality healthcare for patients. PMID- 12764728 TI - [How to improve clinical competence]. AB - Every day there are many patients that suffer or undergo more or less serious damage to their health because of the poor quality of healthcare they receive. Most of these injuries could be avoided by improving the professional quality of healthcare. This presentation outlines an ideal pathway by providing methodological indications and practical tools to clinicians and healthcare workers who wish to increase their skills in planning, supporting and evaluating initiatives of professional quality improvement in their organizations. In particular, the article indicates how clinical competence can be expressed, measured, evaluated and improved. To this end we introduce some basic concepts relative to efficacy (is what I do for my patients useful?), appropriateness (am I doing it to the right persons?) and effectiveness (does the treatment, when transferred to a local operating context, improve the health of the patients receiving it?). The article gives some suggestions to measure quality through process indicators (what has been done to the patients during the process of caring) and outcome indicators (the benefits obtained by the patients from the treatments received), highlighting both advantages and limitations. We then introduce four increasingly complex models for evaluating health interventions: descriptive evaluation, process evaluation, outcome evaluation and experimental designs. Finally, we describe Shewhart's cycle of continuous improvement, which is a useful tool for implementing, directing and spreading continuous quality improvement into an healthcare organisation. PMID- 12764729 TI - [External quality assessment of clinical services in Europe]. AB - All countries and clinical specialties have some elements of systems for quality improvement, but their aims, configurations, models standards and assessments are often not formally recognised or integrated. External programmes to assess service delivery in Europe include the International Standardisation Organisation (ISO) and Excellence (EFQM - European Foundation Quality Management) models (industry based, management focus), peer review and accreditation (health care based, professional focus) and inspection (regulatory, safety-focused). Patient surveys and disease registers also contribute to assessment and benchmarking best performance. There are legal, cultural, professional and commercial reasons to adopt common core standards but there is little legislative framework to allow formal harmonisation within and between countries and clinical specialties. Examples are given of various approaches to the definition, assessment and improvement of standards for clinical services in Europe in order to encourage specialist associations to develop self-regulations based on the experience of others. Clinical practice and clinical services could be more efficiently and effectively harmonised by the professions than by their respective governments. PMID- 12764730 TI - [Quality and efficiency of the Health Service in the Italian Region of Lombardy]. AB - The Regional Law No. 31 of 1997 has revised the Lombardy Health Service as a whole, separating "providers", credited Public and Private Health Institutions, from "purchasers", Local Health Agencies. To this purpose, the improvement of the quality of assistance has been enhanced through an accreditation process aimed at implementing second level regional requirements considered as being necessary for carrying out effective treatment and meeting patient needs. Training, implementation and evaluation of quality service within Health Institutions have been carried out with the assistance of the Regional Observatory on the Quality of Health Service, which helped to identify initial intervention areas, define process indicators, activities and outcomes and verify patient satisfaction, all this to improve performance, reduce risks and control costs. To fully develop the Institutions in the accreditation process, the complex and difficult aspect of health care quality has been more extensively and deeply dealt with by adopting an "Excellence accreditation" model in cooperation with the Joint Commission International Accreditation and by experimenting with Professional accreditation supported by Scientific Societies to further contribute to the development of technical skills and knowledge. The Quality Project carried out so far is based on the integration of various projects, which will be the object of practical interventions by the General Health Direction. All the information and results gathered from this project will be very helpful in finding solutions that will enable all Health Institutions in Lombardy to reach an adequate level of quality. PMID- 12764731 TI - [Institutional accreditation: general observations and local specificities]. AB - The article describes the nature and development of the institutional accreditation process in Italy, with reference to the law in force. This process is an administrative act whose objective is the selection of providers for the National Health Service to ensure quality control and to support quality improvement. The general approach adopted by the Italian region of Emilia Romagna to assess health care facilities is also described. Moreover, the article identifies the role of clinicians and the methods adopted to define the appropriate criteria for specialised contexts. PMID- 12764732 TI - [The institutional accreditation process in the Italian Regions of Emilia Romagna and Lazio: professional contents]. AB - The problems related to the institutional accreditation of the structures of Nephrology, Dialysis and Transplantation has been dealt with in various documents of the Regional Scientific Societies and in Regional Legislative Decrees. Two of these documents are particularly important, one is a special experimental project by the city councillor office for Health Policies issued by the region of Lazio and the other drafted by the Regional Health Agency for the region of Emilia Romagna. What is special about these documents is the fact that nephrologists have actually participated in the drafting process and consequently great attention was given to the professional aspects of the specialised activity. The document issued by the region of Lazio tends to monitor - through criteria, indicators and standards - the various phases of health intervention in Nephrology, by following the different sectors of the process and health care areas identified by the Manual of Accreditation of the Italian Nephrology Society. The document of the region of Emilia Romagna, more complex and articulated, deals systematically with specific requirements (structural, technological, organisational), services, clinical competence and teaching, qualification of diagnostic and therapeutic processes and health care. The Italian Society of Nephrology has been asked to give an official evaluation of both documents. At this particularly uncertain stage, this approach is to be considered as being essential in maintaining consistent performance in the protection of the professional quality of the specialisation in Nephrology. PMID- 12764733 TI - [Regional institutional accreditation measures for nephrology in Italy]. AB - The process of institutional accreditation has been initiated in Italy too because it is considered to be the criterion upon which new relations within the National Health Service must be based. These relations include payment for the service as well as choice of quality assessment and revision of the activities carried out and the services provided. This paper describes the regulations in force regarding Institutional accreditation and the specific regulations of Italian regional credit laws on nephrology. The Regions examined are Lazio, Campania, Marche, Basilicata and Lombardy. A synoptic table showing the most important criteria, differentiated according to each region, is also included. PMID- 12764734 TI - Role of surgery in children with rhabdomyosarcoma. AB - BACKGROUND: Rhabdomyosarcoma (RMS) is a common soft tissue sarcoma of childhood. Historically, surgery has played a central role in the management of children with this tumor, though with surgery alone survival rates were poor. With current multimodal (surgery, radiotherapy and chemotherapy) treatment of these patients, survival has dramatically improved and, with this improvement, there has been an evolution of the role of surgery in the management of this condition. MATERIAL AND METHOD: The contemporary published literature (English) regarding surgical aspects of pediatric rhabdomyosarcoma was reviewed and evaluated. RESULTS: Multimodal therapy has improved the survival of children with RMS from 25% in 1970 to greater than 70% today. Surgical procedures for childhood RMS today are less apt to be exenterative or mutilating than those employed thirty years ago. CONCLUSIONS: Surgery plays a vital role in the diagnosis and treatment of children with RMS. This role has evolved in the context of multimodal therapy and improved survival to an emphasis on less radical procedures with decreased morbidity. PMID- 12764735 TI - Cancer in neonates and infants. PMID- 12764736 TI - PCR detection of clonal IgH and TCR gene rearrangements at the end of induction as a non-remission criterion in children with ALL: comparison with standard morphologic analysis and risk group classification. AB - BACKGROUND: The initial response to induction therapy is currently considered one of the most important prognostic factors in acute lymphoblastic leukemias (ALL). A series of methods for the detection of submicroscopic levels of residual disease in patients with ALL mainly based on PCR and immunophenotyping has been developed, demonstrating that the presence of high levels of residual disease at the end of induction therapy is an important, independent prognostic factor. We determined the usefulness of PCR detection of minimal residual disease using consensus primers as a non-remission criterion. PROCEDURE: Bone marrow samples obtained from 49 children with ALL were analyzed at diagnosis and at the end of induction therapy for the detection of clonal IgH, TCRdelta, and TCRgamma rearrangements by PCR. The results were compared with those obtained by standard morphologic analysis and risk group classification. RESULTS: Patients who had clonality detected at the end of induction showed a significantly higher recurrence rate and lower event-free survival than those without detected clonality (24.9% vs. 89.7%) (P < 0.0001). Multivariate analysis revealed that detection of clonality at the end of induction was the most important, independent prognostic factor when associated with age, number of white blood cells, and immunophenotyping. CONCLUSIONS: PCR detection of clonality using consensus primers is a relatively simple technique that is able to identify patients with a high chance of recurrence, and shows a higher sensitivity and a better prognostic value than standard morphologic analysis and risk group classification, defining a new remission criterion. However, further multicentric prospective studies using this technique employing a larger number of cases are necessary to confirm these findings. PMID- 12764737 TI - Prolylhydroxylase and procollagen type III in long-term survivors of acute lymphoblastic leukemia (ALL): a biochemical approach to HCV-related liver disease. AB - BACKGROUND: We examined two proteins, prolylhydroxylase (hPH) and procollagen type III (PIIIP), as possible non-invasive HCV-related markers of liver disease. The purpose of this study was to assess whether the measurement of these proteins could serve to monitor HCV related liver damage in acute lymphoblastic leukaemia (ALL) patients. PROCEDURE: A total of 34 ALL patients, 24 HCV-seropositive and 10 HCV-seronegative, who had had increased transaminase values (ALT) for almost 6 months were studied. Serum hPH concentrations were determined by an immuno enzymatic assay kit. PIIIP was assayed by the radioimmunoassay method. RESULTS: Both hPH and PIIIP were increased in ALL patients with chronic hepatitis C. Serum hPH levels were significantly elevated in those with chronic hepatitis C with either normal or high transaminases when compared to those who never were HCV seropositive. The sensitivity and specificity of these protein measurements to evaluate hepatic fibrosis were not supported by histologic confirmation because only 6 out of 12 patients with chronic hepatitis had a liver biopsy. CONCLUSIONS: Our study suggests that PIIIP and hPH values are significantly higher in ALL patients with chronic HCV with either normal or high transaminases. This might suggest that the liver damage is more marked in patients with chronic hepatitis and that the liver damage is related to the HCV rather than chemotherapy. Future studies correlating histologic findings with the serum biochemical markers are required to establish the sensitivity and specificity of hPH and PIIIP in predicting hepatic fibrosis and to confirm this association. PMID- 12764738 TI - C-reactive protein (CRP) as tumor marker in pediatric and adolescent patients with Hodgkin disease. AB - BACKGROUND: C-reactive protein (CRP) is an acute phase protein produced in the liver. An elevated CRP is a nonspecific marker of inflammation. Additionally, it also appears to be a prognostic marker in several malignancies. Elevated CRP levels in adult patients with Hodgkin disease (HD) were reported previously. However, levels of CRP have not been evaluated in pediatric and adolescent HD patients. PROCEDURE: We analyzed CRP serum levels in 95 consecutive pediatric and adolescent patients with Hodgkin disease. CRP levels were correlated with stage, absence or presence of B symptoms, and prognosis. RESULTS: At the time of diagnosis increased serum CRP levels were found in 64 % (61/95) of the patients with a median of 21 mg/L (range: <5-211). Serum C-reactive protein levels correlated with stage and were higher in patients with B symptoms. Higher CRP levels were associated with an increased risk of relapse. CONCLUSION: In addition to soluble interleukin 2 receptor (sIL-2R) levels, CRP holds promise as a diagnostic and prognostic index and follow-up monitor in pediatric and adolescent patients with Hodgkin disease, and merits further investigation. PMID- 12764739 TI - Rapid LightCycler assay for identification of the perforin codon 374 Trp --> stop mutation in patients and families with hemophagocytic lymphohistiocytosis (HLH). AB - BACKGROUND: Recently, point mutations in the Perforin gene on chromosome 10q21 have been described to be the cause of hemophagocytic lymphohistiocytosis (HLH) in a subset of patients. Small deletions, missense, or nonsense mutations were found in both coding exons of the gene. One mutation was found apparently independently in different families of Turkish origin. This Trp374stop mutation is located within a cystein-rich epidermal growth factor (EGF) precursor type domain in exon 3 of the Perforin gene. PROCEDURE: To detect this mutation, we developed a rapid "fluorescence resonance energy transfer" (FRET) assay on the LightCycler based on the different melting behavior of mutated and wild-type sequences. RESULTS: In seven out of 20 analyzed patients with defined HLH history, this Trp374stop mutation was detectable within the different genotypes. Additionally, one out of 90 alleles tested in 45 healthy Turkish controls had this mutation. DISCUSSION: We present a rapid and reliable test for the most common Perforin (Trp374stop) mutation in HLH suitable for genetic testing and prenatal diagnosis. The mutation was detected in a large proportion of Turkish patients. Furthermore, a rapid and precise diagnosis of HLH will shorten the initiation of therapy in this subset of patients. PMID- 12764740 TI - FISH analyses for alterations in chromosomes 1, 2, 3, and 11 define high-risk groups in neuroblastoma. AB - BACKGROUND: The prognostic chromosomal markers 1p loss and MYCN amplification (MNA) are only present in a subgroup of approximately 30% of neuroblastomas. To further characterize high and low risk subsets we investigated alterations in chromosome arms 3p and 11q, additional changes in 1p and MYCN as well as the somy status of chromosome 1 in the same sample. PROCEDURE: Fluorescence in situ hybridization (FISH) was used as an alternative technique to PCR/LOH- or comparative genomic hybridization (CGH) analyses. Alterations in chromosomes 3p and 11q were investigated in 182 unselected tumors, 1p loss and MNA in 174 and 179 of these, respectively. The somy-status of chromosome 1 was determined in 165 tumors as it highly correlates with the tumor ploidy. RESULTS: Alterations in the four chromosomal regions were found in the following frequencies: 3p26: 19%, 11q23: 29%, 1p36: 29%, MNA: 19%. Fifty-two percent of all cases displayed structural aberrations in at least one chromosomal region, 83% in stage 4 and 30% in stages 1-3, 4s. All aberrations were thus correlated with stage 4 disease but were also present in a substantial subset of localized and 4s tumors. Trisomy of chromosome 1 was found in 38% of the tumors, disomy or tetrasomy in 62%. Patients with alterations in any of the four chromosomes and di/tetrasomy 1 showed a significantly increased age at diagnosis. Loss in 1p and MNA were closely associated with each other, as well as 3p and 11q aberrations but not the groups 1p/MNA versus 3p/11q. Only a small portion of trisomic tumors showed aberrations in at least one of the four chromosomal regions (14%) in contrast to the majority of the di/tetrasomic cases (74%). As already known the MYCN status discriminated between good and poor outcome in localized and metastatic stage 4 tumors. In addition alterations in 1p or 11q, deletion in 3p and di/tetrasomy 1 were associated with an unfavorable prognosis in MYCN single copy tumors of stages 1 3, 4s. Multivariate analysis revealed 11q alterations and MNA as the most important chromosomal prognostic factors in all stages. CONCLUSION: FISH analyses for chromosomal alterations in 3p and 11q as well as in 1p and MYCN allows to define different groups with an increased risk for disease progression. PMID- 12764741 TI - Impact on survivors of retinoblastoma when informed of study results on risk of second cancers. AB - BACKGROUND: To assess the impact of providing research feedback to retinoblastoma survivors or their parents regarding the risk of second cancers. PROCEDURE: A four-page survey was sent to 801 retinoblastoma survivors and 55 parents to ascertain their reactions to a "results-letter." The "results-letter" provided feedback from a study indicating that retinoblastoma survivors may be at increased risk of second cancers. RESULTS: Three hundred and thirty-nine (339) retinoblastoma survivors and 43 parents responded to the survey. Eighty-four percent (84%) of respondents found the "results-letter" "very" to "extremely" understandable and 72% found it "very" to "extremely" useful. Participants scored "very" to "extremely" to the following emotions: frightened = 28%, anxious = 27%, sad = 25%, overwhelmed = 15%, angry = 11%, and guilty = 6%. Five (1.4%) respondents stated that they would have preferred not to receive the results. Responses did not vary significantly between survivors with different risks of second cancers. No significant differences were observed between males and females. However, parents were significantly more likely to report feelings of anxiety, guilt, anger, being overwhelmed, and frightened compared to adult retinoblastoma survivors (P < 0.05). Individuals with less than a college education were significantly more sad, angry, overwhelmed, and frightened by the information than individuals with a college degree or higher (P < 0.05). Eighteen percent (18%) of all respondents shared the feedback with their physician. The method of choice for receiving results was by letter with contact names and phone numbers. CONCLUSIONS: These findings indicate that research participants want feedback even when the information is upsetting. Additional studies are needed to identify individuals who experience greater levels of distress following feedback in an attempt to provide improved methods of feedback and support. PMID- 12764742 TI - Fatal toxicity following radio- and chemotherapy of medulloblastoma in a child with unrecognized Nijmegen breakage syndrome. AB - BACKGROUND: In large-scale pediatric chemo- and radiotherapy trials a proportion of patients as high as 10-15% is usually reported as having severe treatment related toxicity occasionally resulting in toxic death. Little is known on the underlying predisposition of the individual child. Several hereditary disorders including immunodeficiency (ID) syndromes or repair disorders, Ataxia Telangiectasia (AT), and Nijmegen breakage syndrome (NBS) were associated with an elevated risk for severe treatment related toxicity. PROCEDURE: This report involves the case of a 7-year-old boy with medulloblastoma who suffered from remarkably severe side effects during and after postoperative radio- and chemotherapy. Several months following craniospinal radiation with a total dose of 36 Gy, late normal tissue side effects were observed within the treated volume. Eighteen months after initiation of treatment the patient died due to protracted cardiopulmonary failure. RESULTS: To quantify the intrinsic radiation sensitivity, lymphoblastoid cells were used to examine chromosomal aberrations by fluorescence in situ hybridization detecting between two to ninefold higher chromosomal breakage rates in comparison to cells of average cancer patients. Skin fibroblasts showed in the clonogenic survival assays a twofold increased sensitivity. Western blotting demonstrated a typical lack of Nbs1. PCR-SSCP analysis followed by direct sequencing of positive samples revealed a homozygous truncating mutation of the NBS1 gene (657del5). CONCLUSIONS: This case highlights that severe treatment related complications in pediatric cancer patients may be the result of increased intrinsic radio- and chemosensitivity due to NBS, AT, and other ID syndromes. It is suggested to exclude such conditions in all patients with anthropometric parameters below the 3rd centile and other signs suggestive for repair disorders or ID syndromes. PMID- 12764743 TI - Adult primary extragonadal germ cell tumors: Treatment results and long-term follow-up. AB - BACKGROUND: Primary extragonadal germ cell tumors (PEGCT) are rare neoplasms. They have a poor prognosis, different behavior, and natural course compared to their gonadal counterparts. Both primary and salvage treatment of these tumors constitute a challenge. We retrospectively evaluated the clinicopathologic status, therapeutic implications, and outcome of our patients with PEGCT. PROCEDURE: Between 1991 and 2000, 18 patients with PEGCT (median age 31 years; range 17-63), diagnosed with tru-cut biopsy and treated with cisplatin-based chemotherapy, were evaluated in respect to treatment response and outcome. RESULTS: Cisplatin-based chemotherapy achieved a complete response rate of 27.8% and a partial response rate of 55.5%. Overall response rate was 83.3%. Only three patients were unresponsive to chemotherapy; an additional six patients with residual mass underwent surgical resection and were rendered disease-free by surgery. The 5-year actuarial event-free and overall survival were 63.4 and 71.3%, respectively. CONCLUSIONS: The outcomes of our patients with extragonadal primaries including mediastinal localization appear to be slightly better than those previously reported. Multimodality therapy is essential for these patients and given the relatively poor prognosis, prospective trials with large sample sizes, and new treatment approaches to improve outcome are required. PMID- 12764745 TI - Giant cell tumor of cervical spine in an adolescent. PMID- 12764746 TI - Interferon-alpha therapy, protein kinase C-alpha and Langerhans cell histiocytosis. PMID- 12764747 TI - European training programme in paediatric haematology and oncology. AB - The European Board of Paediatrics has recommended for all paediatricians a three year Common Trunk training in basic paediatrics, followed for tertiary care specialists, by three years of training in the relevant specialty. Paediatric Haematology and Oncology is a complex specialty with a broad range of activities, ranging from the care of children with leukaemia and solid tumours including those of the central nervous system, to all types of non-malignant haematologic disorders and laboratory haematology pursuits. A modular training programme has been recommended to provide a broad standard of training across the whole specialty which should last at least 18 months, and a further period of 18 months that may be used flexibly to prepare trainees for their anticipated future careers. PMID- 12764748 TI - Reflections of an ethicist: research ethics committees and the investigator--a healthy partnership? PMID- 12764749 TI - Trisomy 11 limited to trisomy 21 cells in a mosaic Down syndrome child with acute myeloid leukemia. PMID- 12764750 TI - A homoharringtonine-based regimen for childhood acute myelogenous leukemia. PMID- 12764751 TI - Down syndrome: possible predisposition to retinoblastoma. PMID- 12764752 TI - Solid-pseudopapillary tumor of the pancreas (Frantz tumor) in children. PMID- 12764753 TI - Long-term survival in osteosarcoma patients following retinoblastoma using doxorubicin, cisplatin, and methotrexate. PMID- 12764754 TI - Dramatic response of multiply relapsed hepatoblastoma to irinotecan (CPT-11). PMID- 12764755 TI - Tumor lysis syndrome, neuroblastoma, and correlation between serum lactate dehydrogenase levels and MYCN-amplification. PMID- 12764757 TI - Nodular regenerative hyperplasia of the liver simulating metastases following treatment for bilateral Wilms tumor. PMID- 12764756 TI - Childhood acute lymphoblastic leukemia managed in tertiary care center in a developing country. PMID- 12764758 TI - Cryptococcosis mimicking a pulmonary metastasis in a child with Wilms tumor. PMID- 12764759 TI - Prospective evaluation in children of a new medical device intended to insure the correct administration of intrathecal antineoplastic agents. PMID- 12764760 TI - A teenager with simultaneous Hodgkin disease and thyroid carcinoma. PMID- 12764761 TI - Coexistent neuroblastoma and histiocytosis? PMID- 12764762 TI - Stenotrophomonas maltophilia infection related mortality during induction in childhood acute lymphoblastic leukemia. PMID- 12764763 TI - A lineage switch from AML to ALL with persistent translocation t(4;11) in congenital leukemia. PMID- 12764764 TI - Nephroblastomatosis update. PMID- 12764765 TI - Successful management of a child with clear cell sarcoma of the kidney (CCSK) and multifocal bone metastases at diagnosis. PMID- 12764766 TI - Osteosarcoma in children under three years of age. PMID- 12764767 TI - Transient encephalopathy following high-dose methotrexate. PMID- 12764768 TI - Revised SIOP working classification of renal tumors of childhood. PMID- 12764769 TI - [Preservation of laryngeal function in surgical treatment of hypopharyngeal carcinoma]. AB - OBJECTIVE: To study of the feasibility, the surgical techniques and the results of laryngeal function preservation in surgical treatment of hypopharyngeal carcinoma. METHODS: A retrospective review of 305 (279 males, 26 females, ages ranging from 14 to 77 years) cases with malignant neoplasms of the hypopharynx that were treated from 1978 to 1996 were accomplished. Of the 305 (stage I, 6; II, 12; III, 82; IV, 205) cases, 234 were originated from pyriform sinus, 21 from postcricoid, 35 from posterior pharyngeal wall and 15 from superior hypopharynx. Two hundred and eighty-five patients received 55-75 Gy postoperative radiotherapy. RESULTS: The overall 5 year survival rate of 305 patients was 44.8%; for stage I, the survival rate was 83%; stage II, 71%; stage III, 58% and stage IV, 36.5%. Of the 305 patients, 206 cases (67.54%, stage I, 6; II, 12; III, 65; IV, 123) were surgically treated with laryngeal functions preserved and 99 (32.46%, stage III, 17; IV, 82) total laryngectomy. There were no significant differences in 5-year survival between the functionally preserved group and none functional group. Among the 206 functional preserved patients, 139(67.5%) had all functions preserved (voice, respiration and deglutition) and 67(32.5%) partially preserved (voice and deglutition). CONCLUSION: Only a few patients (31/305, 10%) of hypopharyngeal carcinomas require total laryngectomy, and the preservation of the normal part of the larynx is feasible for the eradication of tumor and preservation of laryngeal function. PMID- 12764770 TI - [Analysis of the late curative effect of 432 cases with advanced squamous cell carcinoma of the maxillary sinus]. AB - OBJECTIVE: To study the late curative effect of different methods used in the treatment of advanced squamous cell carcinoma of the maxillary sinus (SCMS). METHODS: A retrospective analysis of 432 cases with SCMS was done. Among them, 141 cases were in stage III, 291 in stage IV, 244 were male, 188 were female, and the age ranged from 14 to 87 years. Radiotherapy, operation only and the combined treatment (radiation plus operation) were adopted. RESULTS: The 5-year survival rates were 15.6% in radiation group, 21.7% in operation group, and 40.1% in combined group (40% in the preoperative irradiation group, and 43% in the postoperative irradiation group). The 5-year survival rates were 31.7%, 46.6% and 20% respectively for those involving structures above, below or both above and below the Ohngren line. Among the 238 cases with poor curative effect, 198 had local recurrence. CONCLUSION: Combined treatment was the best choice for advanced SCMS. There were no significant differences in the survival rates between preoperative and postoperative radiotherapy groups. The survival rate might be improved by preoperative radiotherapy with a dose of 50 Gy, or postoperative radiotherapy with doses of 60-70 Gy. The prognosis for those with the tumor located below the Ohngren line was much better than those above the line and those both above and below the line. The key to the successful treatment for maxillary sinus carcinoma is to control the local recurrence effectively. PMID- 12764771 TI - [The experimental treatment of laryngeal carcinoma with all-trans retinoic acid]. AB - OBJECTIVE: To investigate whether all-trans retinoic acid (RA) has antitumor effect in vivo. METHODS: Sixteen nude mice were transplanted with human laryngeal cell carcinoma PHC3. Eight of them were injected with RA intraperitoneally and others with solvent as the control. Response was measured and tumor histopathological features were studied. RESULTS: The growth and weight of RA treated tumors were significantly reduced in comparison with those of the control. The growth inhibition rate reached more than 50% in the RA group. Studies also showed that RA-treated cells became well-differentiated. There was a significant decrease of c-myc protein level in the RA treated tumors as compared with the control. CONCLUSION: It is suggested that RA could induce differentiation of human laryngeal carcinoma cells and suppress tumor growth in vivo. Regulation of c-myc gene or protein may be related to those happening. PMID- 12764772 TI - [Local treatment with human laryngeal squamous carcinoma draining lymph node lymphocytes in nude mice bearing human laryngeal carcinoma xenografts]. AB - OBJECTIVE: To increase the number of the tumor draining lymph nodes lymphocytes (TDLNLs) in tumor site and study its anti-tumor effect. METHODS: Interleukin 2 activated TDLNLs were administrated by local injection in 10 nude mice bearing human laryngeal cancer at 10(7) cells/mouse, once a week for two weeks. As a control, 10 nude mice bearing human laryngeal cancer were injected with D-Hanks buffer solution. RESULTS: The volume and weight of TDLNLs treated tumors were significantly reduced in comparison with those of the control (P < 0.05). The observed histopathologic characteristics of treated tumors were tumor necrotic foci and increased infiltration of lymphocytes in tumor and around tumor vessels. Proliferation of spleen nodules was also observed in the treatment group. Remote metastasis was not found in this group. In contrast with the control group, tumor necrosis and proliferation of spleen nodules were not observed and infiltration of lymphocytes in the tumor was not significant. There was a lung metastasis in the control group. CONCLUSION: Local injection of TDLNLs is effective against human laryngeal carcinoma xenografts in nude mice. It also could stimulate the immunoresponse of hosts and probably decrease the chance of tumor metastasis. PMID- 12764773 TI - [The expression in situ of transforming growth factor beta s, their receptors and TGF beta-receptor interacting protein-1 in nasopharygneal carcinoma]. AB - OBJECTIVE: To study the expression of transforming growth factor(TGF) beta s, their receptors and TGF beta-receptor interacting protein-1 (TRIP-1) in nasopharygneal carcinoma. METHODS: Immunohistochemical technology and in situ hybridization methods were adopted to detect the TRIP-1 mRNA and 3 kinds of TGF beta isoforms and 2 kinds of TGF beta receptors protein in the same biopsy specimen. RESULTS: The positive expression of TGF beta 1, TGF beta 2, TGF beta 3, TGF beta R I and TGF beta R II was stronger in the tumor adjacent epithelium than in the tumor itself, which were 65.79%, 66.67%, 55.26%, 48.57% and 63.16% higher than those in the tumor itself respectively(P < 0.01). The level of TRIP-1mRNA measured in the epithelial cells was also higher than that in the tumor cells (19.32 +/- 10.70 versus 11.96 +/- 5.85, P < 0.05). CONCLUSION: The signal transmission of TGF beta family is diminished in the poorly differentiated squamous cell carcinoma of nasopharynx. It may be a factor for the development of nasopharygneal carcinoma. PMID- 12764774 TI - [Intracellular calcium mobilization and effects of ATP on ampullate crista hair cells of the guinea pig]. AB - OBJECTIVE: To further understand the intracellular calcium mobilization features of the isolated ampullate crista vestibular hair cells(VHCs) and ATP effects on intracellular free calcium concentration([Ca2+]i) in VHCs of the guinea pig. METHODS: Vestibular hair cells were isolated from the guinea pig crista ampullaris by enzymatic and mechanical methods. The influence of high K+ solution and ATP on [Ca2+]i was investigated. The [Ca2+]i of VHC was examined using laser scanning confocal microscopy(LSCM) and the Ca2+ sensitive dye Fluo-3. RESULTS: (1) 75 mmol/L K+ Hank's solution resulted in an increase of [Ca2+]i in both the cytoplasm and the cuticular plate of 5 type I VHCs. In the Ca(2+)-free medium, however, 75 mmol/L K+ solution could not induce [Ca2+]i an increase in observed 5 type I VHCs. (2) In the presence of 1.0 mmol/L or 0.1 mmol/L of ATP, there was a rapid reversible rise of the [Ca2+]i in 18/19 type I VHCs. A small response of ATP-induced [Ca2+]i was found only in 1/10 of type I VHC in Ca(2+)-free medium. There was no significant increase in [Ca2+]i when the same volume of Hank's solution was applied to the chamber containing VHCs. CONCLUSIONS: The result suggests that high K+ solution led to Ca2+ influx via voltage-dependent calcium channels. Extracellular Ca2+ influx is a major source of the ATP-induced [Ca2+]i increase. PMID- 12764775 TI - [Effects of sodium salicylate on outward potassium current, resting potential and membrane electric capacitance of the outer hair cell isolated from guinea pig cochlea]. AB - OBJECTIVES: To study effects of sodium salicylate on the outward potassium current (IK), resting potential (RP) and membrane electric capacitance (MEC) of outer hair cells isolated from guinea pig cochleas and the cytoelectrophysiological mechanisms of effects of sodium salicylate on functions of hair cells. METHODS: Using the patch-clamp whole cell recording technique, IK, RP and MEC were measured before and after the administration of sodium salicylate at concentration of 0.1 or 1.0 mmol/L. The effects of sodium salicylate on IK, RP and MEC were observed. RESULTS: The effects of sodium salicylate on IK had time- and dosage-effect relations, of which the former mean IK increasing and then decreasing after administration, the later meant that the effects of sodium salicylate being much more dominant during either increasing or decreasing IK in higher concentration than in lower concentration. The mean RP was approximately- 60 mV and the mean MEC was approximately 39 pF. Sodium salicylate could decrease RP and MEC, which possessed a dosage-effect relation, that is, the effect was stronger in higher concentration than in lower concentration, and there was no clear evidence for time-effect relation. CONCLUSIONS: The sodium salicylate might affect the K+ conduction at the lateral membrane and internal and external distribution of K+ in the outer hair cell. Sodium salicylate might affect the excitability and mechanical activity of the outer hair cell by means of affecting IK, RP and MEC, which might be one of the cochlear mechanisms of the effects of sodium salicylate on the functions of hair cells. PMID- 12764777 TI - [Olfactory evoked potentials produced by electrical stimulation of the olfactory mucosa]. AB - OBJECTIVE: To develop an objective olfactometric method. METHODS: Electrically evoked olfactory potentials were recorded in 20 rabbits with 0.2-4.0 mA electrical stimulations. The stimulating electrodes were placed on the olfactory and respiratory regions of the nasal mucosa. Recording electrode was placed on the scalp near the olfactory bulb. RESULTS: Electrically evoked olfactory potentials composed of triphasic negative-positive-negative peaks, named N1, P1, N2 respectively, were detected on the olfactory region. The latencies of N1, P1, N2 were 16.27 ms, 25.36 ms, 49.75 ms respectively in cribriform plate. No electrically evoked potentials were detected on the respiratory mucosa, nor did on the olfactory mucosa after olfactory neurectomy. CONCLUSION: Steady and clear electrically evoked olfactory potentials which originated from olfactory neurosystem were detected on olfactory mucosa in rabbits. PMID- 12764776 TI - [Protective effect of poly DL-aspartic acid on ototoxicity of gentamicin]. AB - OBJECTIVE: To observe the protective effect of poly DL-aspartic acid (PAA) on ototoxicity of gentamicin (GM). METHODS: Fifty F-344 rats were divided into four groups, GM only PAA + GM, PAA only and saline control. ABR thresholds at different frequencies were measured at different times and hair cell losses were numerated. Two-dimensional diffusion assays in the culture medium were performed to evaluate the effect of PAA on the antimicrobial activity. RESULTS: Eighteen days after the treatment, ABR thresholds at 10 kHz and 8 kHz as well as hair cell losses in the GM-treated group showed significant differences as compared with other three groups (P < 0.01). CONCLUSION: PAA had the protective effect against the cochlear ototoxicity of GM without decreasing its antimicrobial activity. PMID- 12764778 TI - [Effects of rhidosin on expression of fibroblast growth factor on olfactory bulb in senile rats and its significance]. AB - OBJECTIVE: To discuss the effects of rhidosin on expression of fibroblast growth factor (FGF) on olfactory bulb in senile rats and its significance. METHODS: Ten rats each were used in the rhidosin group and physiological saline control group. After decapitation, the olfactory mucosa and olfactory bulbs were immediately fixed with neutral formalin, followed with paraffin-embedding, serial sectioning, immunohistochemical staining and light microscopic observation. RESULTS: The positive expression rate of FGF on olfactory bulb in the rhidosin group was significantly higher than that in the physiological saline control group (P < 0.01). No positive expression of FGF on olfactory mucosa was observed in these two groups. CONCLUSION: Rhidosin can increase the expression of FGF on olfactory bulbs in senile rats and these may be one of the mechanisms by which rhidosin exerts its anti-aging effects on the olfactory system. PMID- 12764779 TI - [Immunohistochemical and pathological study of capsaicin in the treatment of rabbit animal model with allergic rhinitis]. AB - OBJECTIVE: To determine the desensitization effect of capsaicin. METHODS: Toluene 2,4-diisocyanate(TDI) sensitized white rabbits used as animal models of allergic rhinitis were treated with capsaicin. At the end of treatment, the changes in the nasal mucosa were studied by immunohistochemical and pathological methods. RESULTS: Our results showed that the symptoms of allergic rhinitis were remarkably relieved after capsaicin treatment. Immunohistochemical study revealed that substance P immunoreactive fibers in the nasal mucosa showed a marked decrease after capasicin application, but the density and number of SP-IR fibers were significantly increased in the control group. Hematoxylin and eosin staining demonstrated that no edema was found in the nasal mucosa and small vessels were normal after capsaicin application, but edema, vasodilation and inflammatory cell infiltration were discovered in the control group. CONCLUSION: The results suggested that local capsaicin treatment was a selective, and non-traumatic method to induce a long lasting desensitization of the nasal mucosa, to alleviate nasal congestion, rhinorrhea and sneezing, and to reduce the sensory neuron sensitivity of the mucosa. PMID- 12764780 TI - [The analysis of auditory brainstem response during resection of brainstem neoplasm]. AB - OBJECTIVE: To evaluate changes of auditory brainstem response (ABR) and its role in monitoring auditory functions during the brainstem surgery. METHODS: The data of 400 ABRs from 5 operations for removal of astrocytic glioma, cavernomas or dermoid from the brainstem were analyzed. The changes in latencies, amplitudes and wavefroms were dynamically observed and statistically analyzed with t test and Factor analysis. RESULTS: Surgical manipulations, such as stretching, separation and extrusion, induced the prolongation of ABR peaks and interpeak latencies of waves I, III, V. The amplitudes of waves decreased. Three out of 5 cases presented reversible changes in latencies and wave forms. The dynamical analysis showed: The maximal variable range of latency(LMVD) was more than 11.30%, the maximal reversible degree of latency(LMRD) was more than 53.85%, the time of convalescent half-value latency(LCHT) was around 8-40 min, AMVD was more than 40%, the AMRD was more than 200%, and the ACHT was around 5-40 min. In 2 out of the 5 patients the postoperative ABR latencies kept prolonged, some postoperative waves were malformed and difficult to be identified, One case died, another lost consciousness, their LMRDs were less than 33.33%. These findings suggested that surgical intervention had significant effects on the ABR latency (P < 0.01). CONCLUSION: ABR offers a promise for sensitive detection of the brainstem function. Its latency, amplitude and waveform show us dynamic changes of the brainstem status and can be used as an index during the operation on the brainstem and its adjacent region. PMID- 12764781 TI - [The immunological cause of auricular pseudocyst]. AB - OBJECTIVE: To elucidate the relationship between the auricular pseudocyst and the systemic and local immunological status. METHODS: The cyst fluid and blood samples of 44 cases were collected to measure contents of IgG, IgA, IgM and complement C3 by radial immunodiffusion, the content of immune complex by polyethylene glycol turbidimetry, DNA of cytomegalovirus in the fluid by polymerose chain reaction, anti-nuclear antibody(ANA) by immunofluorescent technique and reactions between samples and frozen sections of healthy white rat. RESULTS: Contents of IgG, IgA, IgM and C3 in the cyst fluid were significantly lower than those in the serum (P < 0.01). No immune complex could be detected in both serum and cyst fluid. No DNA of cytomegalovirus was found in cyst fluid, whereas ANA was discovered. Reactive immunocomplex in the auricular cartilaginous tissue was found in the frozen sections of white rats. CONCLUSION: The results suggest that auriular pseudocyst may be an autoimmune disease involving the auricle. The possible mechanism may be an accumulation of reactive exudate due to immunoreactions of autologous antibody and auricular tissue. PMID- 12764782 TI - [Surgical treatment of laryngotracheal stenosis]. AB - OBJECTIVE: To evaluate the surgical treatment results of laryngotracheal stenosis. METHODS: Forty-five patients with laryngotracheal stenosis, atresia or defects who had been treated surgically were reviewed. Different operative reconstructions were used accordingly, such as simple scar resectoin and dilatation, mucosa resuturing, Z-plasty, flap reconstruction, sleeve resection. RESULTS: Forty-three patients had been treated successfully with good airway patency, although 37 of them had hoarseness of voice. One patient lost to follow up and another had compromised airway after operation. CONCLUSION: The different methods of surgical reconstruction used here for the laryngotracheal stenosis proved to be effective. PMID- 12764783 TI - [Immunocytochemical study of gamma-aminobutyric acid-ergic innervation in the end organs of human vestibule]. AB - OBJECTIVE: To investigate gamma-aminobutyric acid-ergic (GABAergic) innervation in the end-organs of human vestibule. METHODS: A modified pre-embedding immunostaining technique of immunoelectron microscopy were applied to accomplish this study with a polyclonal antibody to gamma-aminobutyric acid. RESULTS: GABA immunoreactive products were confined to the nerve terminals, which were rich in synaptic vesicles and the non-myelinated fibers. The GABA-immunoreactive nerve fibers synapse with afferent calices surrounding the type I hair cells. CONCLUSION: This study shows that GABAergic fibers of human vestibular end-organs belong to the vestibular efferent system. PMID- 12764784 TI - [Changes of Ca(2+)-ATPase in the cochlea of guinea pig during labyrinthine hydrop]. AB - OBJECTIVE: To investigate the localization of Ca(2+)-ATPase (Ca2+ pump) in the cochlea and its change in labyrinthine hydrops. METHODS: The left endolymphatic sac was ablated to induce endolymphatic hydrops in fourteen healthy guinea pigs after the sliver ball electrode was placed on the round window. The Ca(2+)-ATPase was studied by the lead citrate reaction in the control and hydropic ears. The reaction product was lead phosphate particles as an expression of Ca(2+)-ATPase activity under the electron-microscope. RESULTS: The Ca(2+)-ATPase activity was found mainly on the endolymphatic surface of the Reissner's membrane, the stereocilia and cuticular plate of inner and outer hair cells, as well as along the infolding plasma membrane of the strial intermediate cells. The Ca(2+)-ATPase activity was significantly decreased during endolymphatic hydrops in the above mentioned locations. CONCLUSION: The response thresholds of filtered click were increased and the Ca(2+)-ATPase significantly decreased in the cochlea during labyrinthine hydrops. These results suggest that correlation exists between the CAP threshold and the activity of Ca(2+)-ATPase in the model of labyrinthine hydrops. PMID- 12764785 TI - [Calcitonin gene-related peptide changes in efferent vestibular system during vestibular compensation]. AB - OBJECTIVE: To investigate the calcitonin gene-related peptide (CGRP) effect on efferent vestibular system during pathological state of vestibular afferent system. METHODS: An animal model of vestibular compensation was made by administration of streptomycin to rat vestibular organ to destroy unilateral vestibular function. Change of CGRP in efferent vestibular neurons in all processes from vestibular disorder to vestibular compensation was observed utilizing ABC method of immunohistochemical technique. RESULTS: Efferent vestibular neurons of normal animals showed low immunoreactivity to CGRP. The number of CGRP immunoreactivity neurons and level of CGRP immunoreactivity increased in efferent vestibular system during vestibular disorder, and these changes decreased with vestibular compensation. CONCLUSION: Activity of CGRP in efferent vestibular system plays a regular role on accelerating vestibular compensation. PMID- 12764786 TI - [Influence of intranasal medication on the structure of the nasal mucosa]. AB - OBJECTIVE: To study the pathological changes and reversibility of the nasal mucosa after drug administration. METHODS: Gentamicin and insulin were dropped into the nasal cavity of rabbits for 3, 5, 7 days to two weeks, after one and two weeks, the nasal mucosa was taken and observed under optical and electron microscopes. RESULTS: It was found that after 3-7 days of drugs administration, damages of the nasal mucosa gradually appeared, and became most serious after one week when the epithelia in some regions detached from basement membr theane. After stopping drug application, the ciliated epithelia restored quickly and completely, goblet cells reproduced and inflammatory reaction disappeared. CONCLUSION: The damages induced by drugs on nasal mucosa are reversible. The intranasal route of drug administration is feasible and promising. PMID- 12764787 TI - [The applied anatomy of the periosteal flap composed of submental artery and dgastric muscle]. AB - OBJECTIVE: To provide anatomical basis for a new style operation using periosteal flap composed of submental artery, anteroventral portion of digastric muscle and submental artery for the renovation of laryngotrachea. METHODS: The anatomical structures correlative with submental artery, submental vein, anteroventral portion of digastric muscle and mandibular periosteum in 23 (46 sides) cephalocervical samples were measured. RESULTS: The periosteal flaps had shown many advantages inculding large transfer capacity, rich blood supply, flexibility. CONCLUSION: It is feasible that the periosteal flap composed of submental artery, anteroventral portion of digastric muscle and mandibular periosteum is useful laryngotracheal renovation. PMID- 12764788 TI - [Bilateral acoustic neuromas]. AB - OBJECTIVE: To investigate the clinical course, growth character and treatment of bilateral acoustic neuromas (BANs). METHODS: Five cases of BANs were reported. RESULTS: From 1986 to 1996, 92 patients with acoustic tumors were managed. Of them, 5(5%) bilateral acoustic neuromas were observed. All patients were males, the age at the time of testing ranged from 16 to 31 years (mean 23.8 years). In all patients, progressive hearing loss and tinnitus were the most common initial symptoms. In addition, signs of other cranial nerve deficits were detected in all patients. Two cases had cafe au lait spots and all cases had neurofibromas of varying sizes and shapes in subcutaneous tissues. Four had unilateral acoustic tumors operated on, one by the translabyrinthine approach, 3 by the suboccipital approach. One patient died due to encephaledema and hernia of brain after operation. According to the MRI data in one case, the average growth rate of acoustic tumor was 1.1 cm (superior/inferior) and 0.7 cm (medical/lateral) per year. CONCLUSION: The management of BANs patients presents challenging problems different from the unilateral acoustic tumors. PMID- 12764789 TI - [Otoacoustic emissions for newborn hearing screening]. AB - OBJECTIVE: To evaluate the validity of transient evoked otoacoustic emissions (TEOAE) and distortion product otoacoustic emissions (DPOAE) in newborn hearing screening and to establish the feasible pass/fail criteria of screening. METHODS: One hundred and eight newborns (216 ears) were screened by TEOAE and DPOAE with Celesta 503 Cochlear Emission Analyzer. The outcome of OAE screening was compared with that of auditory brainstem response(ABR) test. RESULTS: When the criterion of passing TEOAE screening was based on the correlation coefficient of waveforms equal to or greater than 0.60 (r > or = 0.60), the co-positivity and co negativity to the ABR results were 100% and 93.8% respectively. When the pass criterion of DPOAE was set as that the response levels were greater than the 5th percentile (P5) of the normal newborns in more than five out of eight frequencies, the co-positivity and co-negativity were 100% and 94.3% respectively. The outcome of TEOAE and DPOAE screening between r > or = 0.60 and P5 showed the best correlation, r = 0.6583 (n = 216, P = 0.0001). CONCLUSION: r > or = 0.60 and P5 were the feasible pass criteria of TEOAE and DPOAE for newborn hearing screening respectively. OAE proved to be an objective tool for rapid and effective test in universal newborn hearing screening. PMID- 12764790 TI - [Floating culture of human nasal glandular cells]. AB - OBJECTIVE: To establish a floating cultural model of human nasal glandular (HNG) cells. METHODS: HNG cells were cultured on floating collagen gels. RESULTS: Cultured HNG cells incubated with monoclonal antibody for cytokeratin stained positively. After two-week culture, the cells stained blue by AB-PAS staining. Transmission electron microscopy showed that the cells exhibited numerous microvilli on their surfaces and had numerous secretary granules. They were well polarized and interconnected by junctional complexes. Cultured HNG cells possessed typical characteristics of normal epithelium. CONCLUSION: The floating cultural model of HNG was successfully established. PMID- 12764791 TI - [The CT scan of the agger cell and its adjacent structure in endoscopic paranasal sinus surgery]. AB - OBJECTIVE: To promote the development of endoscopic frontal sinus surgery and naso-lacrimal anastomosis surgery by studying the agger cells and their adjacent structures. METHODS: One hundred patients were observed by the continuous coronal CT scan and sagittal CT scan of the paranasal sinuses. Among them, 30 had deviation of nasal septum; 40 had chronic nasosinusitis. There were 80 adult and 20 children. RESULTS: (1) The agger cells existed in 99% of the cases, the development of the agger cells in 9-16 years old children, and adult had no difference; (2) The morphology of agger cells differed widely, but its relationship to the anterior border of the middle nasal concha, bulla ethmoidalis and nasolacrimal canal was quite stable; (3) The upward pneumatization of the agger cells was not related to deviation of nasal septum, but closely related to the development of the frontal sinus; (4) The anterior nasosinusitis often influenced the agger cells and might result in hyperosteogeny. CONCLUSION: The agger cells situated at the pathway of the intranasal frontal sinus during surgery and dacryocystorhinostomy, the nasal CT scan might be beneficial to the prevention of surgical complications. PMID- 12764792 TI - [Possibility and clinical significance of reservation middle turbinate in endoscopic sinus surgery]. AB - OBJECTIVE: To study the possibility and significance or reservation of the middle turbinate in endoscopic sinus surgery for sinusitis and/or nasal polyps. METHODS: (1) The morphology and mucosal ultrastructure of middle turbinate were observed preoperatively and postoperatively by nasal endoscopy and scanning electron microscopy in 20 cases; (2) The proportions of the septal turbinate formation and the closure of ethmoid sinus cavity were investigated postoperatively by nasal endoscopy in 60 cases with middle turbinate reserved and 60 cases with middle turbinate resected. RESULTS: (1) The morphology and mucosal ultrastructures of reserved middle turbinate were observed to have recovered postoperatively; (2) The proportions of the septal formation and the closure of ethmoid sinus cavity in the groups with middle turbinates reserved and resected were 13.3%, 18.3% and 58.3%, 51.7%, respectively (P < 0.01). CONCLUSION: It is possible that the structure of reserved middle turbinate may return to normal and it is important to save the middle turbinates to improve clinical cure rate. PMID- 12764793 TI - [Diagnosis and surgical treatment of meningoencephalocele]. AB - OBJECTIVE: To explore the characteristics of symptoms of meningoencephalocele and the selection of the most appropriate surgical approach. METHODS: From January 1986 to August 1997, 11 cases (frontoethmoidal type 5, basal 5, occipital 1) were retrospectively analyzed. RESULTS: Combined intraextracranial approach was applied 11 times and successful in 10 times; simple frontal approach 1 time, unsuccessful, lateral nasal incision 2 times, failed in 1. No recurrence was found in all successful operations during follow-up. CONCLUSION: Combined intra extracranial approach, which provides excellent exposure and convenient repairing of skull base deficit is an ideal operation for meningoencephalocele. PMID- 12764794 TI - [Morphological study on chronic hypertrophic rhinitis treated with cryosurgery]. AB - OBJECTIVE: To approach the effect of cryosurgery on the mucous membrane of patients with chronic hypertrophic rhinitis. METHODS: Thirty cases of chronic hypertrophic rhinitis treated with cryosurgery were observed by light, transmission and scanning electron microscopy. RESULTS: Before cryosurgery, the mucous epithelia were squamous metaplastic. After the first cryosurgery, the epithelia degenerated and fell off. After the second cryosurgery, the epithelia regenerated and cilia formed. CONCLUSION: The results showed that cryosurgery could diminish the pathological changes and improve the nasal function. PMID- 12764795 TI - [Pharyngoscopic observation during sleep in patients with obstructive sleep apnea syndrome]. AB - OBJECTIVE: To analyse collapse sites of pharynx and their anatomical causes during sleep in patients with obstructive sleep apnea syndrome(OSAS), and compared with the results of Muller's maneuver. METHODS: The pharynges in 43 sleeping patients with OSAS were examined endoscopically. The area changes at each segment(velo-, oro- and hypopharynges) were calculated during the episodes of apnea. A site having greater than 75% narrowing of airway lumen was defined as a narrowing or obstructive site. RESULTS: 1. All patients had obstruction at velopharynx during asleep. Airway collapses at multiple sites in 79% of cases(28% at velo- and oropharynx; 21% at velo- and hypopharynges; and 30% at velo-, oro- and hypopharynges). 2. At each segment the anatomical factors causing obstruction were variable among patients. The common abnormalities were posterior displacement of soft palate or uvula, enlarged tonsil, thicken or posterior displacement of tongue base, collapse of pharyngeal wall, redundant mucosa or lateral bands of pharynx etc. 3. The positivity of pharyngeal obstruction was higher in this study as compared with of Muller's maneuver the latter was taken place while the patient was awaken. Comparison of the positive rate of the narrowing sites determined between sleep observation and Muller's maneuver in same patients: at velopharynx(100% vs 91%), at oropharynx (58% vs 42%) and at hyperpharynx (51% vs 28%). CONCLUSION: Multiple obstructions in the pharynx are common. Examination of the pharynx during asleep may be more accurate then while the patient was awaken. PMID- 12764796 TI - [Studies on localization of laryngeal carcinoma associated antigen by immunoelectromicroscopy]. AB - OBJECTIVE: To study the localization of laryngeal carcinoma associated antigen (LCAA) in the carcinoma tissue. METHODS: Ninety cases of laryngeal carcinoma and 14 cases of laryngeal precancerous lesion and 10 cases of normal laryngeal mucosa were detected with three strains of monoclonal antibodies LC9, LC11, LC12 by immunochemistry. The positive sections of laryngeal carcinoma were observed under light microscope and electromicroscopy. RESULTS: The positive rates of LCAA were dramatically higher than that in normal epithelial and precancerous tissues (P < 0.01). The results showed that the mixed monoclonal antibody had tissue specificity. The MLC associated antigens only distributed in cell membranes and/or cytoplasm. No cell nucleus was stained. CONCLUSION: The LCAA is mainly located in cell membranous structure. This study may provide morphological basis for immunoimaging diagnosis and targeting chemotherapy by application of laryngeal carcinoma McAb. PMID- 12764797 TI - [Studies on treacheostomal formation and voice prosthesis after total laryngectomy]. AB - OBJECTIVE: To establish a tracheostomy model with no stricture and "bowl-shaped" collapse. METHODS: Twenty-five cases were reconstructed with Blom-Singer technique after total laryngectomy for tracheoesophageal voice in our department since 1996. Tracheostoma was done with 5 kinds of methods. In 6 cases, anterior cervical U-shaped incision + division of sternal attachments of bilateral sternomastoid muscles + Griffith tracheostoma enlargement (combination method) were used in the primary operation, and the third generation of indwelling prosthesis and tracheostomal valve were installed. RESULTS: In these 25 cases, 23 could speak freely. The successful rate was 92%. In tracheostoma operation, there were 7 cases with their skin sutured directly to tracheal rings, 3 cases with Myers method, 6 cases with Griffith method, 3 cases with [symbol: see text] method. All of them needed cannulation for more than 6 months and had stricture of tracheostoma and "bowl-shaped" collapse of various degrees. But those underwent "combination method" needed no cannulation and no stricture and bowlshaped collapse was found during 6 mouths' follow-up. CONCLUSION: The combination method of tracheostomy makes the installation, postoperative care and replacement of prosthesis and tracheostomal valve easy and the indwelling prosthesis has the best tone quality. The use of tracheostomal valve effectively makes the patient's hand free during speaking. PMID- 12764798 TI - [Long-term follow-up result of partial laryngectomy in senile patients with laryngeal carcinoma]. AB - OBJECTIVE: To study the long-term follow-up result of laryngectomy with reservation of laryngeal function in senile patients. METHODS: Ninety-nine patients who were above 65 years old and underwent partial laryngectomies from 1986 to 1990 were summarized. Among them, 14 cases underwent vocal cord resection, 31 vertical hemilaryngectomy, 15 horizontal supraglottic hemilaryngectomy, 2 horizontal glottic laryngectomy, 22 horizontovertical (3/4) laryngectomy, 5 subtotal laryngectomy, 10 total laryngectomy with cricopharyngeal anastomosis. RESULTS: All patients had their phonation and respiration restored. Decannulation rate was 91.9%. No death occurred during the operation. No severe postoperative complications were found. 3.5-year survival rates were 76.7% and 75.5% respectively. CONCLUSION: Senility is not a contraindication for laryngectomy with reservation of laryngeal function. The key points are that surgeons should master indications correctly and improve surgical skill to decrease postoperative complications. PMID- 12764799 TI - [Supraglottic horizontal partial laryngectomy]. AB - OBJECTIVE: To evaluate the supraglottic horizontal partial laryngectomy in the treatment of laryngeal cancers. METHODS: Some researches and modifications are performed on the standard supraglottic horzontal partial laryngectomy: the wounded area in the laryngeal cavity is covered with the lateral membrane of thyroid cartilage; the laryngeal cavity is closed as its anterior wall by folding and sewing up the sternohyoid muscles. From 1983 to 1993, 92 patients (80 men and 12 women) with quamous cell carcinoma of larynx were treated with supraglottic horizontal partial laryngectomy. The patients age ranged from 39 to 72 years; 72% aged from 50 to 59. There were 72 cases (78.3%) in the stages III and IV. RESULTS: The 3 and 5 year survival rates of the patients were 75% and 72% respectively. Most cases have acquired good speech and swallow function with 2-4 weeks; 84.8% (78 patients) were extubated. CONCLUSION: The authors believed that partial laryngectomy was safe and beneficial to some patients with laryngeal cancers. PMID- 12764800 TI - [Expression of estradiol receptors in mast cells of the upper airway]. AB - OBJECTIVE: To examine the expression of estrogen receptors in mast cells, CD4+, CD8+ and CD68+ cells in human nasal polyps. METHODS: Immunohistochemical and dual immunostaining methods were used with mouse monoclonal antibodies against human estrogen receptors, a range of human immune cell types included CD4+ (helper T cells), CD8+ (suppressor/cytotoxic T-cells), CD68+ (Macrophages) cells and mast cells. RESULTS: Coincident with the known features of human nasal polyps, all the immune/inflammatory cell types examined were clearly detected with corresponding monoclonal antibodies. The cells positive for estrogen receptor were morphologically similar to the mast cells but distinct from CD4+, CD8+ or CD68+ cells. The cells stained for estrogen receptors by dual immunostaining coincided exactly with cells labeled as mast cells but differed from CD4+, CD8+ or CD68+ cells. CONCLUSION: Estrogen receptors express only in mast cells, but not in CD4+, CD8+ or CD68+ cells in the human upper airway. PMID- 12764801 TI - [Clinical analysis of traumatic optic nerve dysfunction]. AB - OBJECTIVE: To evaluate the key factors in diagnosis, treatment and prognosis of optic nerve trauma at the canalicular segment. METHODS: Twenty-six patients with optic nerve dysfunction resulted from trauma to the nerve at canalicular segment were analyzed. All the patients received orbital CT scans, corticosteroid therapy and surgical optic canal decompression through the external ethmoid approach. RESULTS: With the help of CT scan, the total diagnostic rate was 66.7%; that for canal fracture was 81.3%, for nerve swelling 56.3%. The improvement rate of surgical decompression depended on the time of visual loss and types of toauma. Specifically, the improvement rate was 57.1% in patients with visual loss occurred immediately after the trauma, 83.3% in patients with visual loss occurred a moment after the trauma, 66.7% in patients with optic nerve swelling and 56.3% with optic canal fracture. The possibility of recovery decreased with delay of the surgery. CONCLUSION: Orbital CT scan facilitates the diagnosis of optic canal fracture. However, the possibility of traumatic optic nerve dysfunction can not be ruled out in patients without the sign of canal fracture in the CT scan. The key factors in determining nerve recovery were associated with the severity of optic nerve trauma and the chance of surgery. Serious trauma and delay of the surgery may result in the blindness. PMID- 12764802 TI - [Changes in nasal airway resistance before and after endoscopic sinus surgery]. AB - OBJECTIVE: To study the influence of endoscopic sinus surgery(ESS) on nasal airway resistance (NAR) and to analyze the relationship between NAR and nasal airflow sensation. METHODS: The NAR and nasal airflow sensation were measured with anterior rhinomanometry and visual analogous scale test (VAS) in 42 patients (50 sides) before and after the ESS. RESULTS: NAR decreased significantly after the operation. The change rates in NAR before and after the decongestion were (55.71 +/- 13.90)% and (24.32 +/- 8.73)%, respectively, and there was a significant difference (P < 0.05). The pathological increase in NAR correlated well with subjective sensation of the patients (P < 0.05). CONCLUSION: The improvement of nasal functions by ESS is not due to the alteration of the anatomic structure of the nasal cavity, but to the amelioration of mucosal edema. The rhinomanometry can be used as a useful clinical tool in determining nasal patency. PMID- 12764803 TI - [Anatomic variations of the ostiomeatal complex and their correlation with chronic sinusitis: CT evaluation]. AB - OBJECTIVE: To explore the relationship between anatomic variations in the ostiomeatal complex (OMC) and chronic sinusitis. METHODS: Coronal plane CT scans of the paranasal sinuses showing the ostiomeatal complex including the middle turbinate, uncinate, ethmoid bulla, agger nasi and Haller cell were analyzed in 297 individuals (594 sides). Statistical analyses were carried out with SPSS 5.0. RESULTS: There were 81.14% of OMC noted to have at least one variation. The anatomic variations included paradoxical curvature of the middle turbinate (13.97%), the pneumatized middle turbinate (34.85%), uncinate hyperplasia(19.36%), deviation of uncinate (45.27%), large ethmoidal bulla (30.30%), large agger nasi (0.70%) and Haller cell (1.00%). The correlation between variations and chronic sinusitis was significant (P < 0.01). CONCLUSION: The occurrence of OMC variation is common. The variations may be one of the causes of chronic sinusitis. Therefore, properly handling the variations was important in the endoscopic sinus surgery. PMID- 12764804 TI - [Analysis of HLA-DRB1 allele polymorphism for patients with allergic rhinitis]. AB - OBJECTIVE: To analyze the association between allergic rhinitis and HLA-DRB1 in northeast area of China. METHODS: Using polymerase chain reaction/sequence specific primers (PCR/SSP) technique, we studied distributive speciality of HLA DRB1 alleles in 35 patients with (AR) and 94 healthy controls. RESULTS: The statistical results showed that the frequencies of DRB1 * 0101.2 and DRB1 * 0302 alleles were significantly higher in AR patients than in those of controls. The frequencies of DRB1 * 0101,2 and DRB1 * 0302 were 12.86% (relative risk = 15.92 Pc < 0.0004) and 5.71% (relative risk = 12.00, Pc < 0.05) respectively, the other gene frequencies bore no significant difference. CONCLUSION: AR was associated with HLA-DRB1 * 0101.2 and DRB1 * 0302 alleles. PMID- 12764805 TI - [The protective effects of ciliary neurotrophic factor on inner ear damage induced by intensive impulse noise]. AB - OBJECTIVE: To evaluate the feasibility of using ciliary neurotrophic factor(CNTF) to treat intensive impulse noise-induced inner ear damage. METHODS: The guinea pigs were given either CNTF (CNTF group) or 0.9% sodium chloride (NS group) for 3 weeks after impulse noise exposure. The animals receiving neither medicine nor noise served as a control group. ABR threshold shifts, the cochlear AchE staining as well as the hair cell and spiral ganglion cell counting were carried out in three groups of animals. RESULTS: The numbers of damaged hair cells and spiral ganglion cells in the CNTF group was less than that in the NS group. AchE activity alteration was also less severe in the CNTF group. Similar to the morphological results, changes in the auditory function, represented by the ABR threshold shifts, was less in the CNTF group. CONCLUSION: CNTF can protect cochlear hair cells and spiral ganglion cells against intensive impulse noise exposure by decreasing degeneration and necrosis of the hair cells in some extent and expedite hearing recovery. PMID- 12764806 TI - [Deferoxamine protects against gentamicin ototoxicity]. AB - OBJECTIVE: To study the prevention of gentamicin(GM) ototoxicity by deferoxamine(DFO) in the guinea pig. METHODS: Guinea pigs were randomly assigned to three experimental groups (GM-treated alone, DFO-treated alone, GM and DFO in combination) and one control group. Acoustic brainstem response (ABR), the surface preparation and transmission electron microscopy were utilized to evaluate the hearing thresholds and the cochlear morphology. To explore the mechanism associated with deferoxamine protection, serum levels of GM, BUN and Cr, together with concentration of MDA, SOD and iron in cochlear and renal tissues were measured. RESULTS: The GM group developed up to 40-60 dB of the threshold shifts at 8 kHz while the GM + DFO group developed only 15-25 dB of the threshold shifts (P < 0.05). Moorphological changes were consistent with functional changes. DFO did not alter serum levels of GM. Renal function of GM group was damaged obviously. However, changes of MDA, SOD and iron were not significant (P > 0.05). MDA and iron concentrations in cochlear tissue of the GM + DFO group were significantly lower than those in the GM group (P < 0.05) while SOD level was much higher than that in GM group. CONCLUSION: This study suggests that free radical and iron involve in GM ototoxicity and DFO may become a promising therapeutic agent that can be used to reduce gentamicin ototoxicity. PMID- 12764807 TI - [Morphological observations of the medial olivocochlear efferents in gerbils with chronic gentamicin ototoxicity]. AB - OBJECTIVE: To determine the possible involvement of the medial olivocochlear efferents (MOC) in chronic aminoglycoside ototoxicity, the morphological changes of the MOC efferents in the cochlea of Mongolian gerbils and their correlations with the impairment of the outer hair cells (OHCs) were observed. METHODS: The animals were given gentamicin in a dose of 150 mg/kg a day for 10 days. The distributions and densities of the MOC efferents were examined using the modified histochemical staining for acetylcholinesterase (AChE) and the numbers of OHCs were numerated with toluidine blue and Ehrlich haematoxylin staining on the surface preparation. RESULTS: Significant damage to the MOC fibers and terminals as well as OHCs was noted in the basal turn of the cochlea, especially in those animals examined in 7th and 11th week after the gentamicin administration. The site of the greatest impairment of OHCs was consistent with that of the MOC efferents. CONCLUSION: The results suggested that damage to the MOC efferents might play an important role in gentamicin-induced impairment of OHCs. PMID- 12764808 TI - [Traverse connectivity of innervation in the vestibular sensory epithelium]. AB - OBJECTIVE: To further understand the traverse connections of nerve components in the vestibular sensory epithelium. METHODS: The nerve innervations in the vestibular sensory epithelia and infraepithelia of ten healthy guinea pigs (16 ears) were observed by transmission electron microscopy. RESULTS: Type I sensory cells were intricately connected with type II cells. Afferent and efferent fibers as well as their endings had also complicated connections with nerve trunks and branches. The traverse connections appeared among nerve components in the vestibular sensory epithelium were extremely complicated, forming multilevel connections. They stemmed from and end in the crista ampullaris and the maculae. CONCLUSION: These complicated and multilevel connections are the basis of nerve activity in the vestibular apparatus. PMID- 12764809 TI - [Particle repositioning maneuver for benign paroxysmal positional vertigo of posterior semicircular canal]. AB - OBJECTIVE: To determine the effectiveness of particle repositioning maneuver (PRM) for the treatment of benign paroxysmal positional vertigo (BPPV). METHODS: Thirty one patients with BPPV of the posterior semicircular canal treated between July 1996 and June 1998 were retrospectively analyzed. Patients received a single treatment of PRM based on the hypothesis that the vertigo and nystagmus of BPPV were due to debris floating in the long arm of the posterior canal (canalithiasis). Treatment outcome was classified as cure (asymptomatic), improvement reduction of symptoms > 50%, or no change. RESULTS: After a single treatment of PRM, complete remission of vertigo and nystagmus were found in 67.7% of the patients and improvement of the symptoms was found in 19.4% of the patients. The total improvement rate was 87.1%. CONCLUSION: The particle repositioning maneuver was effective for most of the patients with benign paroxysmal positional vertigo and can be recommended as the first-line treatment modality. PMID- 12764810 TI - [Nasopharygeal size in obstructive sleep apnea syndrome]. AB - OBJECTIVE: To investigate the impact of upper airway 3-D size on the obstructive sleep apnea syndrome (OSAS). METHODS: There were 20 male and 3 female OSAS patients involved in the research. The measured apnea-hypopnea index (AHI) was 45.72(24.01 episodes per hour of sleep). The items measured from magnetic resonance imageing (MRI) of upper airway size were compared to those from 12 normal controls that were sex and age matched to the OSAS patients. The Pearson correlation and multiple linear regression had done to investigate into the relationship of AHI and MRI data. RESULTS: The OSAS patients had relatively narrow nasopharynges than matched controls in the following items: the sagittal size, transverse size, section area and volume. The measured sagittal size of the nasopharynx was closely related to AHI and Lown SaO2, and the volume of nasopharynx was related to apnea index (AI). CONCLUSION: The nasopharyngeal size is closely associated with OSAS. PMID- 12764811 TI - [A clinical and pathological study on cervical lymph node metastasis in the clinical N0 patient with laryngeal carcinoma]. AB - OBJECTIVE: To study the characteristics of the cervical lymph node metastasis in negative nodes (N0) with clinically with laryngeal carcinoma patients and its implication in clinical treatment. METHODS: Forty patients with laryngeal carcinomas of No category were divided randomly into two groups: 13 radical neck dissections (RND) and 27 functional neck dissections (FND) were performed. Lymph nodes were studied histologically. RESULTS: On an average, 34.2 lymph nodes were obtained in one side of neck in RND group, and 27.4 in FND group (t = 0.86, P > 0.05). The metastases rates were 30.8% (4/13) in RND group and 33.3% (9/27) in FND group, and the total metastasis rate was 32.5% (13/40). Twelve of 13 patients (92.3%) who had positive nodes involved only the levels II and III, and 32 of 33 positive nodes (96.9%) were located in the levels II and III. The 3-year survival rates of the two groups were 69.2% (9/13) and 77.8% (21/17), respectively with no statistical difference (chi 2 = 0.3418, P > 0.5). Total 3-year survival rate was 75% (30/40). CONCLUSION: The supraomohyoid (level Io II and III) or lateral neck (level II, III and IV) dissections seem suitable for the treatment laryngeal carcinoma patients with clinically negative neck nodes. PMID- 12764812 TI - [Expression and clinical implication of multidrug resistance gene and multidrug resistant-associated protein gene in patients with hypopharyngeal squamous cell carcinoma]. AB - OBJECTIVE: To study the relationship between the expression of MDR-1 mRNA/P glycoprotein (P-gp). MRP mRNA and the clinical pathological characteristics of the patients with hypopharyngeal squamous cell carcinoma. METHODS: Reverse transcription polymerase chain reaction(RT-PCR) assay was used to detect the expression of MDR-1 mRNA and MRP mRNA in 25 specimens of the untreated advanced hypopharyngeal carcinoma. Meantime, immunohistochemistry SABC technique was used to test P-gp in 60 tumor specimens. RESULTS: The positive expression rates of MDR 1mRNA and P-gp in hypopharyngeal carcinoma were 48 percent (12/25) and 38.3 percent(23/60), respectively. There was a correlation between expression of MDR-1 mRNA and its product P-gp in 25 patients using RT-PCR assay(P < 0.05). The positive expression rate of MRP mRNA was 52 percent (13/25), which was significantly related to metastases of cervical lymph node (P < 0.01). CONCLUSION: Forty-eight percent of the untreated patients with hypopharyngeal carcinoma presented positive expression of MDR-1 mRNA, which was correlated with its gene product P-gp. The expression of MRP mRNA was associated with increased tumor dissemination and metastases. PMID- 12764814 TI - [Advances in basic researches in otology]. PMID- 12764813 TI - [Influence of radiotherapy on reconstruction flaps in oral and maxillofacial regions]. AB - OBJECTIVE: To evaluate the influence of radiotherapy on reconstruction flaps in oral and maxillofacial regions and the radiation tolerance of flaps, and therefore provide clinical information for perioperative radiotherapy. METHODS: Sixty-six free flaps and 22 pedicled flaps were used in the reconstruction of oral and maxillofacial tumor defects. The success and healing of the total 88 reconstruction flaps(14 flaps in preoperative radiation group and 74 flaps in postoperative radiation group) were observed. The acute and late side effects of radiotherapy in the flaps of the postoperative group were studied as well. RESULTS: Of the preoperative 14 flaps, 12 were successful (85.7%), one partially successful and one failed. Of the postoperative 74 flaps, however, 73 were successful (98.6%) and only one partially successful. No total loss of the flap was noted in the postoperative group. The success and healing rates in the preoperative group were lower than those in the postoperative group significantly (P < 0.01-0.05). The rate of acute radiation response of flaps in the postoperative group was significantly lower than that in the surrounding normal tissues(35.1% versus 83.8%, P < 0.01). The late side effects of radiotherapy were rare. The radiation tolerance in different kinds of flaps was similar. CONCLUSION: Reconstruction flaps in oral and maxillofacial regions have a good radiation tolerance and routine postoperative radiation doses can be safely delivered. PMID- 12764815 TI - [Progresses in otoneurosurgery and cranial base sugery]. PMID- 12764816 TI - [Advances in audiology in China]. PMID- 12764817 TI - [Vestibular system diseases: progresses of the clinical and basic researches]. PMID- 12764818 TI - [The challenge of congenital and hereditary diseases]. PMID- 12764819 TI - [Allergic rhinitis: a review of recent literatures]. PMID- 12764820 TI - [Endoscopic sinus surgery: technique and its extensive application]. PMID- 12764822 TI - [Diagnosis and treatment of obstructive sleep apnea syndrome in China]. PMID- 12764821 TI - [Rhinoneurosurgery and rhino-ocular related diseases]. PMID- 12764823 TI - [Advances in basic research for laryngeal cancer]. PMID- 12764824 TI - [Partial laryngectomy: advance and application]. PMID- 12764825 TI - [Differences in furosemide-induced changes of summating potential and endocochlear potential in guinea pigs]. AB - OBJECTIVE: To investigate the physiological properties of summating potential (SP) and to compare the different changes between SP and endocochlear potential (EP) induced by Furosemide in guinea pigs. METHODS: SP in response to clicks with alternative polarities at 105 dB peSPL was recorded from a electrode in the facial nerve canal. EP was measured with a glass microelectrode in the basal turn of the cochlea. SP and EP were observed simultaneously and continuously after injection of Furosemide or the solution of Furosemide-bovine serum albumin injected into the jugular vein of the animal. RESULTS: Before furosemide application, only SP could be recorded with small or without +SP. After furosemide injection, EP declined dramatically followed by reduction of the -SP which quickly vanished, while +SP appeared or the amplitude increased. When EP recovered to a certain level, +SP disappeared or dramatically decreased and then SP reappeared. The amplitude of -SP increased progressively with EP recovery. Both -SP and EP recovered to the pre-injected level in 60 min after the furosemide application. CONCLUSION: The results suggest that Furosemide induced distinct changes of -SP and +SP, the two components of SP. The changes of -SP amplitude was consistent with that of the EP value, while the changes of +SP amplitude were contrary to that of the EP value. PMID- 12764826 TI - [The application of basic fibroblast growth factor to auricles of rabbits with compound injury]. AB - OBJECTIVE: To study the role of basic fibroblast growth factor (bFGF) in the repair of irradiation- and surgery-induced auricle trauma. METHODS: The bilateral auricles of New Zealand white rabbits were received surgical trauma and beta active irradiation(90Sr-90Y), bFGF was then applied to the wound. The efficacy of bFGF on wound healing was compared with the control group. RESULTS: On the 10th day after the trauma, the mean epithelalized area was (34.17 +/- 0.37) mm2 and the mean number of microvessels/high power field was (10.25 +/- 2.34) in the treated group, whereas both were (26.25 +/- 0.20) mm2 and (4.38 +/- 0.92) respectivety in the control group. There was a significant difference between the two groups (P < 0.05). On the 7th and 14th days after the trauma, the mean epithelialized area in the treated group was significantly different from that of the control group. The measurement by the image cytometry showed that the treated group had a mean nuclear peak area value of (38.28 +/- 7.23) micron 2 and the mean S-stage cell value of (31.28 +/- 10.35)%, and those of the control group were (28.02 +/- 8.13) micron 2 and (20.14 +/- 9.46)% respectively. There were significant differences (P < 0.05) between them. CONCLUSION: Application of bFGF can, to a certain extent, accelerate the repair of irradiated and surgically injured soft tissue. PMID- 12764827 TI - [The expression and distribution of ciliary neurotrophic factor in laryngeal nerve regeneration]. AB - OBJECTIVE: To explore the expression and distribution of ciliary neurotrophic factor(CNTF) mRNA and its protein in laryngeal nerve regeneration. METHODS: The recurrent laryngeal nerves were sectioned and then sutured in twelve dogs. Both proximal and distal stumps of sutured region were sectioned on different postoperative days and the sections were separately used for CNTF immunohistochemistry and CNTF mRNA in situ hybridization. The area and intensity of reactive product were measured by computer image processing system. RESULTS: Strongly reactive product of CNTF mRNA and its protein deposited in myelin related Schwann cells in normal laryngeal nerves. At week 2 following neurorrhaphy, there was very little hybridization signal and detectable CNTF immunoreactivity in distal stump and no regenerated nerve fibbers were found. At week 3, reactive product of CNTF mRNA and its protein was detected in thin Schwann cell processes ensheathing regenerated axons, while reactive product was not found in the proliferating Schwann cells which didn't ensheathe axons. CNTF immunoreactivity was also detected in the regenerated nerve axons. After long survival times, hybridization signal and the CNTF immunoreactivity in Schwamm cells became more widespread, and the area and intensity of reactive product significantly increased, but even at the longest survival time, they were still significantly less than those in intact nerve. The same change of CNTF mRNA and its protein was observed in a short segment proximal to the sutured region. CONCLUSION: CNTF expression could depend on Schwann cell-axon regeneration. The level of CNTF expression stands in striking contrast to the up-regulation of nerve growth factor in peripheral nerve degeneration and regeneration. These suggest that the exogenous CNTF might provide a supportive environment for axonal regeneration. PMID- 12764828 TI - [The diagnosis and treatment of bilateral congenital absence of stapes and oval window in two members of a family]. AB - OBJECTIVE: To date, bilateral congenital absence of stapes and oval window in two members (elder brother and younger sister) of a family has not been reported previously in the literature to our knowledge. The etiology, diagnosis and management are hereby introduced and discussed. METHODS: The diagnosis was based on history, audiological examinations, high resolution CT scanning and/or the operation. The modified Lempert's fenestration operation of the horizontal semicircular canal was performed on one ear each of the elder brother and the younger sister. RESULTS: Postoperative hearing threshold was 25 dB for speech frequencies with a gain of 47dB for the elder brother, and hearing threshold of 28 dB with a gain of 52 dB for the younger sister. Following up one to eight years individually showed that their hearing gains were steady. CONCLUSION: Patients' parents are relatives of first cousin and have two sons and one daughter. However, there are two family members suffered from bilateral congenital absence of stapes and oval window, and these malformations might be related to inheritance. Good bone conduction and Carhart's notches revealed bilaterally in the patients' audiograms. These are the important indications for diagnosis of this disease. The modified Lempert's fenestration operation of the horizontal semicircular canal is a safe and good choice for treatment. PMID- 12764829 TI - [Measurement of surfactant in-irrigating fluid from the nasopharynx of patients with secretory otitis media]. AB - OBJECTIVE: Dysfunction of the eustachian tube is believed to play a role in the pathogenesis of secretory otitis media (SOM). In the animal model, previous investigators have found that surfactant in the eustachian tube could decrease the eustachian tube opening pressure and improve eustachian tube functions. Because the nasopharynx is anatomically adjacent to the eustachian tube and the membrane of the nasopharynx continues to the middle ear via the eustachian tube, we hypothesized that the surfactant could also present in the nasopharynx and then designed the following experiment to confirm this hypothesis. METHODS: The concentrations of surfactants in the nasopharyngeal irrigating fluid were measured in normal control and SOM patients. RESULTS: (1) The surfactant from the SOM patients were significantly lower than that from the normal control (8.637 +/ 2.730) mg/L vs. (39.212 +/- 2.437) mg/L. (2) The concentration of surfactant from child SOM was also significantly lower than that of the normal control (8.062 +/- 2.925) mg/L vs. (39.787 +/- 2.557) mg/L. (3) The concentration of surfactant of SOM was not associated with the course of SOM. CONCLUSION: Surfactant reduction in the nasopharynx could indirectly reflect the level of the surfactant in the eustachian tube and the middle ear and influence the eustachian tube function. So surfactant reduction is one of the causative factors in SOM. The strategy of increasing surfactant in the nasopharynx and eustachian tube should be beneficial to reducing persistent secretory otitis media. PMID- 12764831 TI - [Staging of mucous membrane outcome in operative cavity after functional endoscopic sinus surgery]. AB - OBJECTIVE: To investigate the character of transitionary process of mucosa in sinu cavity after functional endoscopic sinus surgery (FESS) and divide it into different stages. METHOD: The morphological character of mucosa in sinus cavity after FESS was surveyed continuously. RESULTS: Over 90% of nasal and sinus cavity got clean within 1 to 2 weeks postoperatively. About 80% of patients had the De mucous reaction from the 3rd to 10th week, such as mucous edema, vesicles, granulation tissue, mini-polyps, fibrous hyperplasia, and adhesion or regenerated diseases may appear in this stage. All of these diseases competed with mucous epithelization. There were 90% of nasal and sinuous cavity brought to stage of complete epithelization, 11 to 14 weeks were need in about 60% of them. CONCLUSION: Three stages were divided after FESS. They are stage of clean operative cavity, stage of mucous competition and stage of complete epithelization. Proper local care in each stage is essential to the whole curable effect of FESS. PMID- 12764830 TI - [Sodium channels in the apical membrane of human nasal epithelial cells]. AB - OBJECTIVE: To study the physiological properties of sodium channels in the apical membrane of human nasal epithelial cells. METHODS: Serum-free cultured nasal epithelial cells of human inferior turbinate from obstructive sleep apnea syndrome patients on collagen gel-coated membranes at an air-liquid interface were studied by patch clamp technique. RESULTS: In cell-attached patches, a typical single channels current was recorded, their conductance was 21.09 pS, the reversal potential was -50.96, and 77.78% of them < -40 mV. Permeability ratio PNa/PK > 5.80. In the presence of 10(-4) mmol/L amiliride in the pipette, the incidence of sodium channels decreased from 26.67% to 5.13%. It revealed that a population of channels were inhibited by amiloride. Ca2+ did not influence the incidence of sodium channels(P > 0.05), there were no obviously association between voltage and the open probability of the channels. CONCLUSION: On Na+ channels in cell attached patches of human nasal epithelial cells, most of channels are amiloride-sensitive and selective Na+ over K+; The channels were not activated by extracellular Ca2+, but the open probability of them were voltage independence. PMID- 12764832 TI - [Immunohistochemical evaluation of micrometastases to cervical lymph nodes in supraglottic cancer]. AB - OBJECTIVE: Supraglottic carcinomas of larynx tends to metastasize to cervical lymph nodes. The regular pathological method of diagnosis is easy to have the micrometastasis missed, and serial-sectioning is too difficult be used in the regular pathological examinations. The aim of this study was to evaluate the value of immunohistochemical assay in the detection of micrometastasis. METHODS: Monoclonal antibodies against cytokeratin (AE1/AE3) were used to detect the cervical lymph nodes of 46 cases (50 sides of neck) of supraglottic carcinoma with pathologically negative findings (pN0). RESULTS: Micrometastases were identified in 13 lymph nodes of 11 cases. CONCLUSION: It is suggested that the detection of lymph node micrometastases with immunohistochemical method is more sensitive than that of regular pathologic method. It is comparable to serial sectioning. Therefore, It is a valuable clinical method to detect the early lymph node metastasis. PMID- 12764834 TI - [Laryngeal function preserving surgery in hypopharyngeal carcinoma]. AB - OBJECTIVE: To investigate the results of the conservation laryngectomy in patients with hypopharyngeal carcinoma. METHODS: Twenty-three cases with hypopharyngeal carcinoma that were treated with function-sparing laryngectomy during a 15-year period were reviewed retrospectively. RESULTS: The 3-year survival rate was 73.9% (17/23) and 5-year survival rate was 47.8% (11/23). Severe complication occurred in 8 of 23 (34.8%). Complete restoration of laryngeal function was achieved in 20 cases (87%) and deglutition restored in all patients (100%). The mean duration of oral intake was 11.8 days (6-33 days) and mean hospitalization was 32 days (14-94 days). CONCLUSION: Conservation laryngectomy and hypopharyngectomy is not only efficacious in the treatment of hypopharyngeal carcinomas, but also reserves laryngeal function and thus improves the quality of patients life. PMID- 12764833 TI - [The reconstruction of laryngeal function in subtotal laryngectomy with preservation of arytenoid cartilage]. AB - OBJECTIVE: To evaluate subtotal laryngectomy with preservation of arytenoid cartilage to reconstruct laryngeal function in T3 glottic or supraglottic laryngeal cancer. METHODS: Subtotal laryngectomy with preservation of arytenoid cartilage to reconstruct laryngeal function was reviewed in 20 patients with T3 laryngeal cancer(3 cases with glottic cancer and 17 cases with supraglottic cancer). Recurrent laryngeal nerve was preserved during surgery. The mucous membranes between cricoid cartilage and pharynx were sewed up accordingly. RESULTS: All patients had restored phonation and swallowing functions. The 3- and 5-year survival rates were 94.1% and 91.8% respectively. Decannulation rate was 95.0%. CONCLUSION: Subtotal laryngectomy with preservation of arytenoid cartilage is recommended for T3 laryngeal cancer. PMID- 12764835 TI - [Laryngotracheobronchial involvement in relapsing polychondritis]. AB - OBJECTIVE: To investigate the clinical cause, treatments and prognosis of laryngotracheobronchial involvement in relapsing polychondritis (RP). METHODS: Thirteen patients of RP with laryngotracheobronchial involvement were review retrospectively. RESULTS: From 1983 to 1998, 20 patients with RP were managed in this hospital. Of them, 13 (65%) involving the laryngotracheobronchial region were observed. Respiratory tract lesions might be diffuse or localized and occurred in larynx (46%), trachea (69%) and bronchus (69%). Corticosteroids, immunosuppressive agents and dapsone were used in these patients. Tracheotomy was performed in 4 patients with airway obstruction. Montgomery T-tube or self expanding metal stent were used in 2 patients with tracheal collapse. Of 13 case, 2 died from respiratory complications. The 5-years survival rate was 78.6%. CONCLUSION: Respiratory tract involvement is the most severe manifestation of the relapsing polychondritis and the main cause of death despite aggressive medical therapy or tracheotomy. PMID- 12764837 TI - [Conservative surgery for glottic cancer with vocal cord fixation (T3)]. AB - OBJECTIVE: To study the method of reconstruction of laryngeal function by unilateral bi-pedicled and bi-muscular flaps for glottic cancer of T3 category. METHODS: From 1984 to 1992, the unilateral bi-pedicled and bi-muscular flaps were used in 68 patients with T3 glottic cancer for repairing the surgical defect, enlarging the piriform recess on the affected side and closing the anterior wall of the laryngeal cavity after vertical laryngectomy. RESULTS: The 5-year survival rate was 73.5% (50/68). The decannulation rate was 91.2% (62/68). sixty-three patients had satisfactory phonation (92.6%). All patients resumed mouth-food taking within 7 to 20 days after operation. CONCLUSION: The unilateral bi pedicled and bi-muscular flaps are appropriate for reconstruction of surgical defect in T3 glottic cancer. PMID- 12764836 TI - [Surgical management of subglottic carcinoma]. AB - OBJECTIVE: To explore the clinical features and the methods of surgery and functional restoration of subglottic carcinoma. METHODS: Thirteen cases with primary subglottic carcinoma were treated surgically in this department from 1981 to 1997. Among them, six had T1-2N0 lesions, one had T3N0 lesion and six had T3 4N1-2 lesions. Four cases underwent total laryngectomy and nine had subglottic partial laryngectomy. The extensive subglottic partial laryngectomy was performed on patients with tracheal invasion. The defects of larynx were reconstructed by using unilateral or bilateral pedicled musculocutaneous flap, myofascial flap accordingly. T-shape silastic tube was placed in to the reconstructed cavity of larynx during the operation and the patients were decannulated in 2 to 6 months. Unilateral radical neck dissection was performed on 4 patients and bilateral on 2. RESULTS: All cases had restored the function of phonation except for 4 who underwent total laryngectomy. Five out of 9 (55.6%) were decannulated. The swallowing function was restored in all patients. The 3-year and 5-year survival rates were 100% and 66.7% in the cases with total laryngectomy, 88.9% and 75.0% with subglottic partial laryngectomy, respectively. CONCLUSION: It is possible to detect early subglottic carcinoma by using fiberoptic laryngoscopy routinely with the combination of stroboscopy, CT, MRI in male patients over 40 with hoarseness. It is practical that the whole or partial function of larynx could be restored in most cases with subglottic carcinoma after partial laryngectomy or subtotal laryngectomy. Unilateral or bilateral neck dissection should be performed on patients with T3 or T4 lesion. Postoperative radiotherapy is necessary. PMID- 12764838 TI - [Supracricoid partial laryngectomy for laryngeal cancer]. AB - OBJECTIVE: To analyze the key points, especially the indications and contraindications of supracricoid partial laryngectomy (SCPL). METHODS: During the years 1995 to 1996, 18 patients with laryngeal cancer underwent supracricoid partial laryngectomy. Ten of them (55.56%) were classified as T1b and T2 glottic cancers, 8 of them (44.44%) as T3 and T4 glottic cancer or transglottic carcinomas. Three types of supracircoid partial laryngectomy (CHP, CHEP and TCHEP) were employed accordingly. RESULTS: Overall three-year survival rate was 94.40%. All patients were decannulated from 14 to 42 days and all resumed normal oral feeding from 14 to 30 days. Speech was good in all patients. CONCLUSION: The SCPL procedure not only increases local control rate, but also maintains physiologic speech and swallowing. PMID- 12764839 TI - [Minimally invasive treatment in early glottic cancer and vocal cord dysplasia]. AB - OBJECTIVE: To investigate the pathologic and clinical diagnosis of early stage glottic carcinoma and the vocal cord dysplasia and the results of the minimally invasive treatment. METHODS: Thirty-two cases of early glottic cancer (T1N0M0) and 20 cases of vocal dysplasia were treated with mucous stripping and minimally invasive cordectomy under the micro-suspension laryngoscope. The pathomorphology was observed with continuous sections. RESULTS: After 3-4 years follow-up, 26 cases of early glottic cancer remained tumor free, 3 cases relapsed and 2 cases received the re-stripping operation. twenty cases of vocal cord dysplasia were classified as: leucoplakia of vocal cord, mild atypical hyperplasia, moderate atypical hyperplasia and advanced hyperplasia. None of them had malignant change after 3 years follow-up. CONCLUSION: The laryngeal micro-operation under the micro-suspension laryngoscope is an effective and safe cure procedure, it provides a definite diagnostic method for the early stage glottic carcinoma and the vocal cord dysplasia. PMID- 12764840 TI - [Modification of cochlear active micromechanism by auditory cortex]. AB - OBJECTIVE: To investigate the modification of active micromechanism of outer hair cells by auditory cortex. METHODS: 1. Effects of selective attention on distortion product otoacoustic emissions (DPOAE) were analyzed in 9 human subjects with normal hearing. Visual tasks, two letters, O and Q, were alternatively displayed on a computer screen. Letter Q was target stimulus flashing randomly (probability was 20%), and letter O was non-target stimulus occurring regularly (probability was 80%). 2. In 13 guinea pigs, DPOAEs were measured before and after decerebration. RESULTS: 1. Visual tasks suppressed DPOAE significantly in both 1 kHz and 2 kHz (P < 0.05). 2. DPOAE reduced significantly in decerebrated guinea pigs. CONCLUSION: The study further demonstrates that the efferents from the auditory cortex do innervate the cochlea, and the efferent pathway passes from the auditory cortex, the super olive complex to the cochlea. Through this pathway the auditory cortex modulates the cochlear active micromechanism. The underlying mechanism was discussed. PMID- 12764841 TI - [The effects of salicylate on noise-induced hearing loss in the guinea pig]. AB - OBJECTIVE: To investigate the effects of salicylate on noise-induced hearing loss (NIHL) in guinea pigs. METHODS: The animals were allocated into four groups (salicylate + noise, saline + noise, noise only and salicylate only) and were exposed to a 4 kHz octave band noise at 105 dB SPL for 2 hours for 5 consecutive days. Dynamic changes of ABR thresholds evoked by clicks were monitored. After physiological examination, the cochleae were processed for the morphological examination by light microscopy. RESULTS: Mean thresholds of ABRs in the experimental group obtained on 5th day of the noise exposure was positively lower than the control groups (P < 0.05). The similar trends of ABR changes were found during other exposure days and after exposure, but the differences were not statistically significant. In the experimental group, there were 37.00 +/- 8.89 hair cell loses as assessed by nucleus staining, and 3.20 +/- 1.07 hair cells with nuclear deformation. The noise group had 74.11 +/- 10.28 hair cell loses and 10.43 +/- 9.71 hair cells with nuclear deformation, whereas the saline group had 63.60 +/- 7.97 hair cell loses and 20.80 +/- 15.48 hair cells with nuclear deformation. The difference between two groups was significant (P < 0.05). CONCLUSION: The results suggest that administration of salicylate facilitates the recovery of cochlear hearing and reduces damage to hair cells after noise exposure. PMID- 12764842 TI - [The biological behavior of jugular glomus tumor and the perioperative management]. AB - OBJECTIVE: Based on our experiences, the biological behavior and perioperative management of the jugular glomus tumor are introduced and discussed in order to prevent the possible mortality and morbidity. METHODS: Five cases of jugular glomus tumor treated in our institute were reviewed. Laboratory examination, modern imaging techniques such as CT, MRI, 131I-MIBG, DSA, as well as embolization of the blood vessels and/or antihypertensive drugs were selectively used in the preoperative period. RESULTS: Surgical removal of the tumors was successful in this group. CONCLUSION: Perioperative management plays an important role in the prevention of surgical complications. PMID- 12764843 TI - [Auditory neuropathy]. AB - OBJECTIVE: To study the clinical feature and audiological and electrophysiological characteristics of auditory neuropathy. METHODS: The history, audiometry, stapedial reflex, electrocochleogram, auditory brainstem response, promontory stimulation tests, otoacoustic emissions (OAE) and its contralateral supression tests were studied in 17 cases of auditory neuropathy. RESULTS: The patients were predominently female teenagers with a history of gradual onset of bilateral hearing loss, pure tone auditometry revealed symmetric ascending audiograms with moderate hearing loss at frequencies below 1 kHz, but relatively normal hearing at 2 kHz and above. Speech reception thresholds were between 40 to 85 dB HL, discrimination score was from 29% to 68%. Stapedial reflex, auditory brainstem response and compound action potentials were absent or threshold elevated disproportionately to the pure tone threshold. All of the patients had normal cochlear microphonics and robust spontaneously and transiently evoked otoacoustic emissions (SOAE and TEOAE), the TEOAE showed no contralateral suppression effects as seen in normal subjects. CONCLUSION: The diagnosis of the disease depends on the discrepancies between subjective and objective hearing and pre-neural and neural responses. The crossed olivocochlear bundle function testing in terms of contralateral acoustic stimulation induced otoacoustic emission reduction is specific for differential diagnosis. There is a need for more awareness of the disease among otolaryngologists and neurologists. PMID- 12764844 TI - [The significance of matrix metalloproteinases expression in nasal polyps]. AB - OBJECTIVE: To explore the role of matrix metalloproteinases (MMPs) in nasal polyp formation. METHODS: The expression of MMP9, MMP2 and MMP3 was studied in 32 nasal polyps by immunohistochemical method. RESULTS: MMPs could not be detected in all samples of normal mucosa. In nasal polyp tissue, MMP9 was observed epithelial in cells, inflammatory cell and blood cells, MMP2 was detected in fibroblasts, epithelial cells and inflammatory cells, and MMP3 was detected in inflammatory cells and epithelial cells. Compare to normal mucosa, MMP9, MMP2 and MMP3 was over expressed in their positive cells of polyp. The expression of MMP9 was higher than that of MMP2 and MMP3 in nasal polyp tissue (P < 0.01). CONCLUSION: MMP9, MMP2 and MMP3 were strongly correlated with nasal polyp formation. They cause pathological changes in nasal mucosa and MMP9 may play the most important role among them. PMID- 12764845 TI - [The correlative factors and preventive management of synechiae following endoscopic sinus surgery]. AB - OBJECTIVE: To study the correlative factors of synechiae following endoscopic sinus surgery and their clinical significance. METHODS: Thirty-two cases (47 sides) of surgical cavity synechiae occurred in 338 (513 sides) endoscopic sinus surgeries were analyzed. The types of synechiae and correlative factors such as pathological features, primary ailment severity, pneumatization of middle turbinate, middle turbinate management, surgical procedures and follow-up management were analyzed. RESULTS: 1. The synechiae occurred in 9.16% of cases. 2. The synechiae occurred successively as: ethmoid sinus cavity synechiae and occlusion, synechia between middle turbinate and lateral nasal wall, middle turbinate synechiae with nasal septum, inferior turbinate synechiae with nasal septum. There was no synechiae between inferior turbinate and middle turbinate. 3. Synechia frequently occurred in patients with inflammatory lesions, severe primary ailment, pneumatization of middle turbinate, pathological middle turbinate, improper surgery and follow-up management and delayed follow-up. CONCLUSION: It is very important to fully recognize every correlative factors of synechiae pre-operatively in order to reduce synechiae. In addition, improvement of surgical technique, appropriate management of middle turbinate, in time follow up and separation of synechiae timely are also indispensable to successful surgery. PMID- 12764846 TI - [EB virus infection and expression of P53 and P16 in non-Hodgkin's lymphoma]. AB - OBJECTIVE: To observe the presence of Epstin-Barr virus (EBV) and expression of P53 and P16 in the non-Hodgkin's lymphoma (NHL) occurring in the nasal cavity and pharyngeal Waldeyer's ring. METHODS: In situ hybridization was used to detect EBV DNA and P16-mRNA and immunohistochemical staining was used to examine the expression of LMP1, P53 and P16 in the 36 cases. Five cases whose EBV-DNA were negative at first assay by hybridization was re-detected for EBV-DNA by PCR. RESULTS: Positive expression rates of EBV-DNA, LMP1, P16 mRNA, P53 and P16 were 63.9%, 27.8%, 30.6%, 19.4% and 22.2%. P53 and P16 expressions were not significantly different between T cell NHL and B cell NHL, nor were them significantly different between EBV positive and negative cases. CONCLUSION: A higher percentage of EBV infection was detected in the nasal cavity and pharyngeal Waldeyer's ring in NHL. EBV infection was not significantly associated with P53 and P16 expresions in NHL. PMID- 12764847 TI - [Expression of CD44v and nm23-H1 proteins in laryngeal carcinoma and its correlation with clinical significance]. AB - OBJECTIVE: To investigate the correlation between the expression of two gene proteins and their clinical significance in laryngeal carcinoma. METHODS: Sixty fresh laryngeal carcinoma samples were analyzed and 20 para-carcinoma tissue and 12 normal laryngeal mucosa samples were studied as controls. All specimens were collected from patients operated on from 1996 to 1998 in the same department (including 52 males and 8 females, age ranged from 41 to 77). The quantitative expression of CD44v and nm23-H1 proteins was analyzed by flow cytometry (FCM) and immunofluorescence technique. RESULTS: 1. Higher level expression of CD44v and nm23-H1 proteins was detected in the laryngeal carcinoma specimen than in normal laryngeal mucosa. 2. The expression of CD44v protein was positively correlated with the metastasis, clinical staging and pathological classification; but not correlated with sex, age, tumor size, course of the disease, T classification and category of laryngeal cancer. 3. The expression of nm23-H1 protein was inversely correlated with metastasis and clinical staging while positively correlated with age and tumor size; no significant relations were found between the expression and the sex and course of the disease, T classification and category of laryngeal cancer. 4. There was an inverse correlation between the protein expression of CD44v and nm23-H1. CONCLUSION: CD44v and nm23-H1 probably play an important regulating role in the metastasis of laryngeal carcinoma. PMID- 12764848 TI - [Endoscopic CO2 laser arytenoidectomy in the treatment of bilateral vocal cord paralysis]. AB - OBJECTIVE: To evaluate endoscopic CO2 laser arytenoidectomy(ECO2LA) for the treatment of bilateral vocal cord paralysis (BVCP). METHODS: Eight cases have been operated upon and observed for 6 to 40 months. Three of them were extralaryngeal approach artenoidectomy failures. All of them had tracheostomy before. The technique was to modification of Ossoff ECO2LA model. A small part of ventricular fold and posterior part of vocal cord was vaporized by laser. The mucoperichondrium overlying the arytenoid was reserved and sutured. After 3 months the opposite arytenoid was treated similarly. RESULTS: There were no aspiration and no granulation at the surgical site postoperatively. All cases kept satisfactory voice. Five of 8 patients were decannulated. Two patients still had laryngeal stridor during sleep and they had to plug the tracheotomy tube only on daytime. One patient had cannula retained for waiting operation on the opposite side. CONCLUSION: ECO2LA is currently a most reliable and efficient technique for treatment of patients with BVCP. PMID- 12764849 TI - [The treatment of stricture of larynx and trachea with Ni-Ti shape memory alloy stand]. AB - OBJECTIVE: To evaluate the use of Ni-Ti shape memory alloy stand in the treatment of stricture of larynx and trachea. METHODS: The clinical materials of 14 patients were analyzed. All patients had stricture of larynx and trachea and accepted the treatment with Ni-Ti shape memory alloy stand. RESULTS: All patients breathed freely after the operation, and the trachea fistula was closed at the time of operation. The follow-up survey for these patients lasted for 2 to 12 months. None of the patients had signs of the stricture of the respiratory tract. The short-term effective rate was 100%. CONCLUSION: Ni-Ti shape memory alloy stand can expand the stricture of larynx and trachea quickly and thus improve the breath effectively. It can be used as a routine method in the treatment of stricture of larynx and trachea. PMID- 12764850 TI - [The effects of substance P antibody on tuning curve of auditory evoked potential of cochlear nucleus and inferior colliculus in guinea pigs]. AB - OBJECTIVE: To explore the effects of substance P on the frequency analysis of sound signals in the auditory brainstem. METHODS: The electrodes were implanted into the cochlear nucleus (CN) and the inferior colliculus (IC) of guinea pigs. The auditory evoked potential in CN (CN-AEP) and IC (IC-AEP) was recorded with tone pip stimulation and tone burst forward masking techniques. The Q10 dB values of the tuning curves were compared between the CN-AEP and IC-AEP after injection of SP antibody or rabbit serum with the same dosage (used as control) into the nucleus. RESULTS: After injection of SP antibody, the Q10 dB values of the tuning curves of CN-AEP and IC-AEP had no significant changes in 1 kHz and 2 kHz probe tone groups. When probe tone frequencies were 4 kHz and 6 kHz, there was a significant difference between the two frequency groups in Q10 dB. In addition, the higher frequency slope and lower frequency slope value had corresponding changes. CONCLUSION: The results imply that SP could act as a neuromediator in the auditory afferent system and participated in the high frequency analysis in the auditory brainstem. PMID- 12764851 TI - [The expression of Ras protein in human middle ear cholesteatoma]. AB - OBJECTIVE: To evaluate the possible role of Ras protein in acquired middle ear cholesteatoma. METHODS: The specimens from the acquired middle ear cholesteatoma tissue of 22 cases and the normal external ear skin of 10 cases were examined by immunohistochemical S-P method and computer image analysis. RESULTS: All epithelial layers of cholesteatoma revealed a relatively abundant expression of Ras protein, among which 15 cases showed an inner plasma membrane expression, 5 cases a cytoplasm and membrane expression, and 2 cases cytoplasm expression only. In contrast, the normal external ear skin revealed a sparse distribution of Ras protein in the inner plasma membrane of epithelial cells with a more intensely staining in the basal layer. The integral absorbency of Ras protein in the two types of tissues were 0.870 and 0.463 respectively with a significant difference between them. CONCLUSION: There is an aberrant regulation of Ras protein in cholesteatoma. Ras protein plays a pivotal role during the aberrant growth and the differentiation of cholesteatoma. PMID- 12764852 TI - [Audiologic analysis of a family with nonsyndromic genetic progressive sensorineural hearing loss]. AB - OBJECTIVE: To analyze the audiological data collected from a large six-generation family with nonsyndromic genetic progressive sensorineural hearing loss. METHODS: There were 104 members in the family who could provide the hearing data, in which 46 individuals had hearing impairment. The pure tone threshold in 56 individuals and ABR in 49 individuals were analyzed. The pedigree of 104 family members with 6 branches was made. RESULTS: Hearing impairment was found in the second decade. The hearing thresholds of high and mid frequencies were deteriorated up to 110 dB HL in the fourth decade. The hearing loss started from high frequencies and quickly expended to mid and low frequencies. The regression analysis of pure tone thresholds of 4 kHz and 8 kHz in 6 cases with ages from 10 to 20 years were carried out. The regression coefficients, or the annual threshold increases were 15.96 dB and 15.34 dB per year, respectively. ABR results showed that the hearing loss was caused by cochlear impairment. The pedigree of family indicated high or complete penetrance. CONCLUSION: The medical history and audiological analysis of the family members showed that this hearing impairment was nonsyndromic autosomal dominant progressive sensorineural hearing loss. The hearing impairment and onset age in this family were different from those families with 13 loci related autosomal dominant nonsyndromic sensorineural hearing loss reported previously. The genetic linkage study showed that it could not link to these regions and other two regions reported recently in Internet. PMID- 12764853 TI - [Analysis of surgical reconstructions of canal and middle ear via "direct entrance" approach in 53 ears with congenital aural atresia]. AB - OBJECTIVE: To recognize the indication of surgical reconstruction via the "direct entrance" surgical approach (superoanterior surgical approach) for congenital atresia of the external acoustic canal and malformations of the middle ear. METHODS: A series of 53 ears operated on via the superoanterior surgical approach during 10 years from January 1988 to December 1997 was analyzed. RESULTS: The tympanic cavity was encountered without difficulty in 52 ears. Facial paralysis occurred postoperatively in one ear, which recovered after two months. The hearing improvement was observed in 45 ears (84.9%), of which 24 ears had hearing improvement greater than 30 dB and the air-bone conduction gap was within 15 dB (45.3%). A long-term (1-9 years) follow-up of 25 ears demonstrated that the hearing levels in 18 ears were kept unchanged. Postoperative stenosis of the canal occurred in one ear of a 2 years-old child. CONCLUSION: Based on the scanning images of the temporal bone by high-resolution computed tomography before the operation, the authors classified the bony structures of the paths of external acoustic canals into four types. Our results indicated that the selerotic and the diploetic paths of external acoustic cmeati could be indications for the "direct entrance" surgical approach, whereas the mixed type, either containing a little small cells or diploetic bone in sclerotic bone, might be suitable for this surgical approach also. PMID- 12764854 TI - [Rate discrimination and tone recognition in mandarin-speaking cochlear-implant listeners]. AB - OBJECTIVE: To explore the relationship between the rate discrimination and the tone recognition with psychophysical method, and provide a basis for evaluating the results from mandarin-speaking cochlear implantees who used multi-channel cochlear implant and designing new speech coding strategies of the speech processor. METHODS: Four postlingually-deafened adults with cochlear implant participated in this study. We used the method of constant stimuli to measure the rate discrimination as a function of the standard rate at 50, 100, 200, 400 Hz on electrode pairs (E1, E3), (E7, E9), (E14, E16), and (E20, E22), respectively. We also measured tone recognition in 25 consonant-vowel syllables, each of which has four tonal variations, resulting in a total of 100 words. These words were recorded by a male talker and were presented five times each to the implant users via their speech processors. The tone recognition results were scored in overall percent correct and analyzed by error patterns in the confusion matrix and fundamental frequency. RESULTS: The rate discrimination thresholds, defined at the 75% correct level, varied greatly among individuals and ranged from several Hz to hundreds of Hz. A three-way ANOVA indicated significant effect (P < 0.05) on rate discrimination for the standard rate, electrode position, and patient variable. The tone recognition ranged from 40% to 80% and tones 1 and 3 were more easily identified than tones 2 and 4. Preliminary analysis using linear regression revealed highly significant correlation between rate discrimination and tone recognition. CONCLUSION: These results suggest that identification of tones relies on good rate discrimination in cochlear implant listeners. Clinical rehabilitation strategies in improving rate discrimination were discussed. PMID- 12764857 TI - [Transtemporal and transinfratemporal combined approaches in lateral skull base tumor surgery]. AB - OBJECTIVE: Safe and complete removal of the tumor invading lateral skull base and its surrounding structures. METHODS: Tumor removal through combined approach of temporal and infratemporal fossa with microsurgical techniques for preservation or reconstruction of lateral skull base and cranial nerves. RESULTS: Sixty-two cases of benign and malignant tumors undergone surgery and followed up for five and more years showed: no recurrence in 67.7%, survival with tumor 25.8%, death 6.5% (only for malignancy). No complications of extra- or intra-cranial infection, cerebrospiral fluid leak and flap necrosis. CONCLUSION: Combined approach of temporal and infratemporal fossa is suitable for the surgery of those tumors that involve temporal bone and middle or posterior intracranial fossa, as well as infratemporal fossa. PMID- 12764855 TI - [Congenital atresia of the external auditory canal]. AB - OBJECTIVE: To investigate the clinical characters and treatments of congenital aural atresia. METHODS: From 1986 to 1996, 44 patients (50 ears) with congenital aural atresias were operated in the ENT Department of the PLA General Hospital. Postoperative audiological follow-up ranged from 6 months to 9 years. RESULTS: Ossicular chain anomalies were presented in all cases. This consisted most frequently of fusion (54%) or deficiency (36%) of the malleus and the incus with or without fusion to the tympanic wall. Facial nerve abnormalities were seen in 16 ears (32%). After the operation, hearing improvement was found in 33 ears (66%), in which 13 ears (26%) had hearing improvement (pure tone threshold) up to 40 dB. The postoperative complications were external auditory canal stenosis, lateralization of the tympanic membrane and recurrent infections of the cavity and canal skin, with incidences of 34%, 18% and 14%, respectively. CONCLUSION: Maintenance of width of the aural canal and prevention of lateral healing of the transplantated tympanic membrane were crucial in the treatment of congenital aural atresia. PMID- 12764856 TI - [Study on the relationship between Chlamydia infection and otitis media with effusion]. AB - OBJECTIVE: To evaluate the relationship between Chlamydia infection and otitis media with effusion and to establish a method for detection of Chlamydia antibodies in local fluids. METHODS: Sera and tympanocentesis aspirates from 44 patients with otitis media with effusion were collected. The specimens were used to detect the Chlamydia specific IgG, IgM antibodies using micro-immune fluorescent assay; to detect Chlamydia trachomatis plasmid DNA and Chlamydia pneumoniae genome DNA with polymerase chain reaction (PCR); and to separate Chlamydia pneumoniae in tympanocentesis aspirates with Hep-2 cell culture. RESULTS: One Chlamydia pneumoniae positive culture was found in tympanocentesis aspirate culture from one of the 44 patients, with positive Chlamydiae pneumoniae specific IgG antibody and DNA. Chlamydia trachomatis specific IgG antibody was increased in 14 cases sera and local secretion accordingly, with Chlamydia trachomatis plasmid DNA positive by PCR. CONCLUSION: Chlamydia is a common pathogen in otitis media with effusion. PMID- 12764858 TI - [Management of middle turbinates in endoscopic sinus surgery]. AB - OBJECTIVE: To explore the relationship between the partial middle turbinectomy and the prognosis. METHODS: Forty patients with nasal polyp and sinusitis were chosen. The partially middle turbinate was removed on one side and preserved on the opposite side. The time of postoperative debridement for nasal cavity, synechia of nasal cavity, and olfactory changes were assessed. RESULTS: The time of postoperative debridement of nasal cavity in patients with partial endonasal middle turbinectomy was shorter than those with preservation of middle turbinate (P < 0.01). There was no synechia in cases with partial middle turbinectomy and 12 cases of synechia in cases with preservation of middle turbinate. There was not difference about olfactory change in both sides. CONCLUSION: The partial middle turbinectomy in the endoscopic sinus surgery may be good for the prognosis of patients. PMID- 12764859 TI - [Continuous positive airway pressure therapy after uvulopalatopharygoplasty]. AB - OBJECTIVE: To investigate if uvulopalatopharygopalasty (UPPP) compromised continuous positive airway pressure (CPAP) therapy. METHODS: Automated continuous positive airway pressure(Auto CPAP) was applied to treat 15 patients with obstructive sleep apnea syndrome(OSAS) whose apnoea-hypopnoea index (AHI) was still higher than 20 after UPPP, and also it was used in the treatment of 32 randomly selected OSAS patients who had not had any treatment before. The treatment results of these 2 groups were compared. RESULTS: It was found that AHI and apnoea index(AI) between the 2 groups after Auto CPAP treatment were not significantly different (P > 0.05), however, lowest SO2(LSO2) after Auto CPAP treatment of the UPPP group was higher than that of the other group (P < 0.01). The AHI, AI and LSO2 in each group were significantly different before and after Auto CPAP treatment (P < 0.01 in both cases). CONCLUSION: It is suggested that CPAP remains effective after UPPP and UPPP doesn't compromise CPAP therapy. PMID- 12764860 TI - [A study on the treatment of congenital velopharyngeal insufficiency]. AB - OBJECTIVE: To evaluate the effectiveness of speech therapy after pharyngoplasty in patients with congenital velopharyngeal insufficiency (CVPI). METHODS: Seven patients with CVPI had received pharyngoplasty and speech therapy. The blowing test and the Chinese articulation clear degree test were measured before and after speech therapy. RESULTS: The treatment results of seven patients appeared to be reasonably satisfactory after speech therapy. Both the index of blowing test and the Chinese articulation clear degree test increased significantly after speech therapy. CONCLUSION: Pharyngoplasty combined with speech therapy is effective in the treatment of patients with CVPI. PMID- 12764861 TI - [Study on the association between squamous cell carcinoma of larynx and HPV subgene by PCR]. AB - OBJECTIVE: To measure the human papilloma virus(HPV) HPV-E1 in squamous cell carcinoma of larynx. METHODS: Polymerase chain reaction(PCR) was used to detect the HPV subgene in 25 specimens of laryngeal carcinoma and 6 normal larynges. RESULTS: Positive expression of HPV-E1 was detected in 17 of 25 cases(68%) with squamous cell carcinoma and all the recurrent cases. However, there was no positive expression of HPV-E1 in the normal larynges (P < 0.01). The expression rate was not significantly different among the histological grad groups. CONCLUSION: HPV-E1 infection was found in laryngeal carcinoma but not related to its histologic grading. PMID- 12764862 TI - [Cervical lymph node metastasis and recurrence in supraglottic carcinoma after operation]. AB - OBJECTIVE: To evaluate the regional control efficacy of elective neck dissection (END) in patients with supraglottic carcinoma. METHODS: The incidence of cervical recurrences of 582 patients with supraglottic carcinoma treated in this hospital from 1981 to 1993 were retrospectively reviewed. RESULTS: Three hundred and ninety-two cases (67.35%) of this group had T3 and T4 primary lesions. Among them, 147 cases (37.50%) had positive nodes(N+) and this accounted for 86.47% (147/170) of all N+ patients. Ipsilateral or bilateral cervical recurrence occurred in 33 of 126 (26.19%) node negative (N0) cases who had not had neck dissection and 30 of them (90.90%) were among T3 and T4 groups. However, Contralateral lymph node recurrence occurred only in 40 of 286(13.99%) N0 cases who had had END and 30 of them (75%) were among T3 and T4 groups. There were 163 cases who were readmitted and received surgery again for different purposes. The three and five year survival rates in the second stage END group were 86.67% and 77.78%, in metastasis group 64.81% and 32.50%, in recurrent group 33.33% and 30.77% respectively. There were significant differences in survival rates among these groups. CONCLUSION: Patients underwent elective neck dissection achieved better survival than those on "wait and watch" policy and salvage surgery. Elective neck dissection is recommended for the treatment of supraglottic carcinoma, especially for the T3 and T4 diseases. PMID- 12764863 TI - [Pathologic feature of occult lymphatic metastasis in supraglottic carcinoma]. AB - OBJECTIVE: To study the pathologic features of occult lymphatic metastasis in patients with supraglottic carcinoma and its implication in treatment. METHODS: Serial sections of 153 neck dissection specimens in 100 patients with supraglottic carcinoma were studied under microscope. RESULTS: Clinically negative nodes had been proved histologically positive in 58 specimens (38%) and 51 occult nodes, size ranged from 0.5 cm to 2.6 cm with average of 1.1 cm. The distribution of metastastic lymph nodes was 1 in level I (2%), 37 in level II (73%), 12 in level III(23%) and 1 in level IV(2%). Among the 51 nodes, 21(41%) were in early stage, 18 (35%) in growth stage, 7(14%) in full stage, 5(10%) were extracapsular. CONCLUSION: Occult metastasis rate of supraglottic carcinoma is as high as 38%, selective neck dissection is recommended for the surgical treatment of this disease. PMID- 12764864 TI - [Posterior sigmoid sinus approach glossopharyngcal neurotomy in the surgical treatment of glossopharyngeal neuralgia]. AB - OBJECTIVE: To appraise the diagnotic criteria and surgical indications of glossopharyngeal neuralgia. METHODS: Under 2% lidocaine local infiltration anesthesia, retrosigmoid sinus approach was adopted to find the cerebellopontine angle. The possible focus causing glossopharyngeal neuralgia was searched carefully under the microscope and the glossopharyngeal nerve cut. RESULTS: Only 2 cases had artery compression on the surface of glossopharyngeal nerve during the operations. Besides, all of them had arachnoid synechia around the glossopharyngeal nerve. The symptoms of glossopharyngeal neuralgia disappeared thoroughly after operations. Three cases having histories of bradycardia, arrhythmia or hypotension recovered from their diseases postoperatively. Recurrent symptoms were not found in the follow-up survey which lasted 8 months to 6 years, with an average of 3.7 years. CONCLUSION: Glossopharyngeal neurotomy via retrosigmoid sinus approach has positive therapeutic effect as long as the diagnosis is correct. PMID- 12764865 TI - ["Antibiotic resistance"]. PMID- 12764866 TI - [MRSA/MRSE-VISA/GISA/VRSA-PRP-VRE: current gram positive problem bacteria and mechanism of resistance, prevalence and clinical consequences]. AB - The raising frequency of occurrence of multi-resistant Gram-positive pathogens has led to difficult to treat infections not only in hospitals but also in outpatient settings. Methicillin-resistant Staphylococcus aureus, the glycopeptide-susceptibility of which one cannot be sure of any more, vancomycin resistant enterococci and penicillin-resistant pneumococci are on the top of the list of Gram-positive problem organisms nowadays. Clinical microbiology laboratories are faced with the challenge of accurately detecting emerging antibiotic resistance, which might be difficult and purpose of reporting data should be twofold: First to report adequately identified, relevant organisms as well as their susceptibility profiles to clinicians in context with adequate patient management and secondly to report continuously epidemiological data in form of statistics to the community in general and within a given setting as could be a specified hospital in particular. For reliable detection, laboratories may need to employ special screening- and susceptibility testing methods. Certain of these tests are highly specific, while others may require additional confirmatory testing for definite results. This article reports on current resistance mechanisms of gram-positive pathogens and subsequently resulting consequences for clinicians. Moreover, the frequency of occurrence in Austria in general as well as at the Vienna General Hospital in particular will be referred to. PMID- 12764868 TI - [Antimicrobial combination therapy]. AB - Anti-microbial combination therapy is a very frequently used therapeutic approach although clinical data from prospective randomized trials thus fulfilling the criteria of evidence based medicine are very rare. Rationales for the use of combination therapy are thus empirical treatment of polymicrobial infections, infections in critical ill or in immuno-compromised patients, prevention of the emerge of antibiotic resistances, utilization of a potential synergism of antibiotics directed against a certain bacterial strain, or simply despair. Prospective studies have demonstrated a superiority of combination therapy over single drug regimen in enterococcal endocarditis, and infections with gram negative rods including pseudomonas sp., but in the latter condition only if the patient is severely immuno-compromised. There is clinical indication that combination therapy may be beneficial in infections with staphylococci sp., however, well designed studies supporting this experience are largely absent. Nonetheless, antimicrobial combination therapy also harbors some risks for patients which can be referred to potential antagonism between antibiotics, accumulation of side effects, or stimulation of resistance pathways in bacteria by one drug leading to destabilization of the other one. Thus, the insightful use of combination therapy is warranted and well designed, prospective studies are urgently needed to evaluate the true benefit of combination therapy for the treatment of different infections and specific micro-organisms. PMID- 12764867 TI - Occurrence of Salmonella enterica serovar Dublin in Austria. AB - In Austria, Salmonella enterica serovar Dublin, a bovine-adapted serovar, rarely causes human infections. In the year 2000, Austria was within the European mean with an incidence of 0.1 per million inhabitants. Our data show that the vast majority of all Austrian serovar Dublin infections can be traced to two Tyrolian districts. This concentration of cases can be explained by a particularly traditional aspect of cattle farming in Tyrol, the alpine pasture. There is increased risk of cross-infection due to the communal keeping of animals from various farms. Infected cattle are a source of infection for people, and contagion usually occurs from eating beef and drinking cow's milk. Using pulsed field gel electrophoresis and automated ribotyping, 3 out of 5 available isolates from human infections could be traced to characteristic Tyrolian S. Dublin clones. Bacteriological screening of herds with a known history of S. Dublin infection would be a start to prevent future contamination of alpine pastures through latently infected cattle excreting potentially infectious feces. Bacteriological screening for fecal carriage before the return of cattle from pastures known to be connected with infections could prevent cross-contamination of large mixed herds. PMID- 12764869 TI - The influence of comorbidity on the effect of levofloxacin treatment success of ambulatory respiratory tract infections. AB - The influence of patient relevant parameters such as age, comorbidity, or duration of disease on the treatment success of levofloxacin for community acquired respiratory tract infections (CARTI) has not been thoroughly elucidated. We therefore conducted a prospective cohort study of 9831 patients with CARTI in a clinical practice setting. The patients received 500 mg of levofloxacin once a day over a mean of seven days. Twenty-two attributes per patient were recorded before treatment initiation and after seven to fourteen days after start of treatment. Descriptive and explorative statistics such as the k-means and C4.5 algorithms were used to analyze the dataset. The overall success rate of levofloxacin therapy for CARTI was over 98%, side effects occurred in 1.6% of patients. Descriptive analysis revealed a weak correlation between parameters which significantly influence the course of disease, such as the number of comorbidities, the duration of infection before levofloxacin start, or the severity of symptoms and the treatment success. Explorative statistics yielded similar results. Two homogenous clusters, holding 34 and 45% of patients respectively, yielded the number of comorbid conditions and the duration of infection as main attributes negatively influencing treatment success. We therefore conclude, that the number of co-morbid conditions and the duration of infection before start of treatment as the strongest negative predictors for treatment success. PMID- 12764870 TI - Determination of the frequency of inadequate antibiotic therapy using calculation of indication failure (IF), cumulative indication failure (CIF), and balanced indication failure (BIF). AB - Because results of pathogen identification are often lacking when antibiotic therapy is initiated, treatment must frequently be instituted on an empirical basis. The type of empirical therapy will depend on the anticipated pathogen spectrum and naturally also on the prevailing resistance patterns. Inadequate antibiotic therapy may not only be associated with increased overall treatment costs, but will also have adverse effects on mortality. The clinician is frequently faced with an overabundant variety of microbiological data and may fail to interpret them correctly. Therefore, the present study has attempted to "translate" the available microbiological resistance data, frequently presented in the form of percentage rates, into concrete patient numbers and thus illustrate the frequency of inadequate antibiotic therapy. For this purpose, "Indication Failure" (IF), "Cumulative Indication Failure" (CIF) and "Balanced Indication Failure" (BIF) have been calculated based on available microbiological data. For the indication "nosocomial pneumonia", calculations of the BIF show that only one out of 67 or one out of 63 patients is inadequately treated with a therapy with cefepime or imipenem, while one out of 25 patients is inadequately treated when using ceftazidime. However, it must be pointed out that these calculations only represent an interpretation of microbiological data and the success of antibiotic therapy will ultimately also depend on parameters such as the pharmacodynamic properties of an antibiotic or on the immunocompetence of the patient treated. PMID- 12764872 TI - ["Psychotherapeutic medicine"]. PMID- 12764871 TI - [New antimicrobial drugs: an update]. AB - Several procedures such as antimicrobial restriction, microbial monitoring, calculated antimicrobial treatment and the development of new antimicrobials are used to reduce the resistance. The following reviews deals with new substances such as carbapenems, quinolones, ketolides, oxazolidinones, glycylcyclines, echinocandines and azoles. PMID- 12764873 TI - [Position of sexual medicine in medical specialties]. AB - The position of sexual medicine among the different medical subspecialities is discussed. Sexuality is seen under its socio-communicative, reproductive and lust aspects which in turn influence specific characteristics of sexual medicine, as e.g. its biopsychosocial way of thinking, its focus on the couple, the alloying of different forms of therapy and especially the stressing of the communicative meaning of sexual behaviour as a form of non-verbal communication. Hence therapeutic interventions aim at the restoration of the sexual relationship as a means of fulfilling fundamental needs and desires as for acceptance, warmth, security and so on and thus exceed the cure of the sexual dysfunction. Sexual Medicine should be integrated into the study of medicine as a subject of its own. PMID- 12764874 TI - [Hypnosis in medical psychotherapy practice]. AB - As doctors we are required to pursue symptom complaints and treat them, no matter whether they bear objectively verifiable evidence of specific disorders or not. In treatment we also take Hypnosis background feelings, conflicts or fatigue into account as contributing components. This fact pertaining to the practice of medical psychotherapy is central to the treatment of all patients suffering from psychosomatic disorders--i.e. approximately one half of all patients being treated by general practitioners. Here hypnosis and self-hypnosis can serve well in the formulation of therapeutic goals as well as in diagnostic clarification. Hypnosis is above all else useful in encouraging patients to contribute their very own individual resources to a multi-modal and interdisciplinary comprehensive therapy concept. This proves itself in the long run to be a factor that promotes a significant emancipation of the patient from those dependencies that are an outgrowth of illness (see hypnosis and self-hypnosis for symptom control) as well as the treatment system (optimal coping, self management) through the application of personal skills. A bio-psycho-social understanding of disorder is--both for ourselves as medical doctors (as "specialists of the objective dimension of the body") and for our patients (as "specialists of the subjective dimension of person")--a "conditio sine qua non". PMID- 12764875 TI - ["Without song or sound?" Possibilities and effects of music therapy]. AB - Music therapy is still not widely known within everyday medical and psychotherapeutical contexts. Music touches fundamental themes of human existence and is important in preventive as well as in clinical and rehabilitative areas. The highly specialized and technical areas of medicine need to be complemented by artistic therapeutic approaches that recognize the physical and sensory aspects of the suffering person. Active and receptive music therapy and training have evolved in different fields of health care. A theory embracing various approaches is particularly useful. Fundamental attitudes in the music therapy relationship will be differentiated and the importance of music therapy will be presented using examples based on the treatment of cancer patients. PMID- 12764876 TI - [Frames of reference for interdisciplinary management in human sciences]. AB - The reference framework for those disciplines that deal with the functions of the nervous system has a very simple basic structure: It becomes clear when, based on the matrix with the four central questions of biological research (causation, ontogeny, adaptation, phylogeny), one asks and at the same time takes the reference levels (e.g. cell, organ, individual, group) at which the questions are aimed into account (the questions and planes in bold are also the subject of the humanities). This "bio-psycho-social" orientation framework is the basis for the development of an interdisciplinary consensus: It is the starting point for a systematic order for those disciplines, and also the basis for a consistent networking and structuring of their results. PMID- 12764877 TI - [Time--the 4th dimision in medicine and psychotherapy]. AB - Time is presented as well in his historical meaning and as 4th dimension in its medical and psychotherapeutic context. In this medical and psychotherapeutic process it has an important function and is a variable of a process procedure. The difference between "kairos" = (the right point of time) and "chronos" = (the period of time) is historically meanful. The subjective experienced time is as well emphasized by the development of time in the relation to the development of the "self" as in the subjective experience of time in medical and psychotherapeutic situations. There are also changed conceptions and understandings of time running parallel to the development of nature sciences. The importance of time is explained for the medical practice and the meeting with the patient--especially for chronic diseases. The connection of confidence and time is particularly emphasized in the systemic approach. PMID- 12764878 TI - Responding to the call of professionalism. PMID- 12764879 TI - Where is the virtue in professionalism? PMID- 12764880 TI - Educating for professionalism: what counts? Who's counting? PMID- 12764881 TI - Fostering professionalism: the Loyola model. PMID- 12764882 TI - Mapping, modeling, and mentoring: charting a course for professionalism in graduate medical education. PMID- 12764883 TI - The other side of professionalism: doctor-to-doctor. PMID- 12764884 TI - CQ sources/bibliography. PMID- 12764885 TI - Human experiments and national security: the need to clarify policy. PMID- 12764886 TI - Conceptions of family-centered medical decision making and their difficulties. PMID- 12764887 TI - Management mistakes in healthcare: a disturbing silence. PMID- 12764888 TI - Helping clinicians find resolution after a medical error. PMID- 12764889 TI - Ward ethics: "What do I do now?" "Who am I?". PMID- 12764890 TI - The case: can doctors say "enough"? PMID- 12764891 TI - How to manage hot flashes in prostate cancer? PMID- 12764892 TI - When privacy and the public good collide. Does the collection of health data for research harm individual patients? PMID- 12764893 TI - Probing rosacea's mysteries. PMID- 12764894 TI - Nonsustained ventricular tachycardia: another approach. PMID- 12764895 TI - The evolution of treatment guidelines for diabetic nephropathy. Strategies integrate JNC VI, more recent protocols. AB - In recent years, new drug development and late-breaking research data have put treatment guidelines for diabetic nephropathy in a state of flux. In particular, trials of angiotensin-converting enzyme (ACE) inhibitors and angiotensin II receptor blockers (ARBs), coupled with enhanced understanding of the renin angiotensin pathway, have influenced recommendations for patient care. Here, Dr Bakris highlights both the steadfast features and the recent refinements of treatment guidelines for diabetic nephropathy. He describes their backing in research findings and outlines practical antihypertensive and renoprotective therapies to curtail risks of nephropathy and cardiovascular disease in patients with diabetes. PMID- 12764896 TI - Managing thyroid dysfunction in the elderly. Answers to seven common questions. AB - Despite the high prevalence of thyroid dysfunction in the elderly, there is often considerable delay and difficulty in diagnosis because symptoms are subtle and attributed to normal aging. Atypical presentations also are common. Routine screening of asymptomatic, healthy adults is not recommended; however, physicians should maintain a high index of suspicion and have a low threshold for testing thyroid function. Subclinical abnormalities of thyroid function are even more prevalent than overt disease. Although observational studies have shown that subclinical disease affects function, adverse clinical outcomes have not been demonstrated. In addition, the benefits of treatment of subclinical disease are unclear. Therefore, treatment should be individualized and restricted to high risk patients. The elderly are vulnerable to the side effects of thyroid replacement and antithyroid medications, which mandates cautious use of drugs and careful dose adjustments. PMID- 12764897 TI - Defining diffuse Lewy body disease. Tetrad of symptoms distinguishes illness from other dementias. AB - It is an ageless principle in medicine that physicians cannot diagnose and treat an illness they do not recognize. In the past two decades, we have come to recognize the classic clinical picture of diffuse Lewy body disease. Once primary care physicians understand this challenging disorder, they are in an ideal position to identify and manage it. As with many dementing illnesses and other medical conditions, diffuse Lewy body disease is not curable, but much can be done to improve the quality of life of patients and their family. PMID- 12764899 TI - Pityriasis rosea. Appearance and distribution of macules aid diagnosis. AB - A 40-year-old man presented with a 1-week history of a diffuse pruritic rash. The patient denied having had fever, chills, or a recent respiratory infection. He could not recall having an initial patch. Physical examination revealed numerous small, scaly patches on his trunk (figure 1). The rash spared the palms of his hands and soles of his feet as well as the distal extremities. PMID- 12764898 TI - The growing problem of pressure ulcers. Evaluation and management for an aging population. AB - Pressure ulcers, an important concern in older adults with restricted mobility, promise to become an even bigger issue as the US population ages. These ulcers can lead to devastating complications and place demands on an already stressed healthcare system. They also can be a quality indicator of the preventive measures taken in healthcare facilities. In this article, Drs Dharmarajan and Ahmed present guidelines for the prevention and treatment of pressure ulcers. PMID- 12764900 TI - Patient notes: macular degeneration. PMID- 12764901 TI - [Concept and methodology of clinical trial randomization]. PMID- 12764902 TI - [Designing and applying of the placebo-needle in clinical trial of acupuncture]. PMID- 12764903 TI - [Study on treatment of myelo-dysplastic syndrome by intergrative traditional Chinese and Western medicine]. PMID- 12764904 TI - [Clinical study on treatment of chronic aplastic anemia assisted by TCM for reinforcing shen and activating blood circulation to dredge channels]. AB - OBJECTIVE: To observe the clinical efficacy of TCM for reinforcing Shen and activating blood circulation to dredge Channels in auxiliary treating chronic aplastic anemia (CAA), and to explore its mechanism. METHODS: Seventy-nine patients with CAA were randomly divided into the treated group treated with TCM plus western medicine and the control group treated with western medicine alone. The clinical efficacy was observed and levels of plasma sFas and sFasL before and after treatment were determined by ELISA. RESULTS: The basic cure rate and total effective rate in the treated group were 37.50% and 87.50%; while in the control group were 20. 51% and 61.54% respectively. Comparison between the two groups showed significant difference (P < 0.01). The sFas level was markedly lower in both groups before treatment, but after treatment, it got elevated in the treated group significantly (P < 0.05), as compared with that in the control group, the difference was significant (P < 0.01). CONCLUSION: TCM is effective in treating CAA with less toxic and side-effect, the mechanism might be the inhibition on over apoptosis of hematopoietic cells in CAA patients through regulating immune function of organism. PMID- 12764905 TI - [Study on efficacy of treatment of acute leukemia by shengfu injection in combination with chemotherapy and the effect on cellular immunity, serum interleukin-6 and tumor necrosis factor-alpha levels]. AB - OBJECTIVE: To observe the efficacy of treatment of acute leukemia by Shengfu Injection (SFI) in combination with chemotherapy and the effect of treatment on T lymphocyte subsets (CD4, CD8, CD4/CD8), serum interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha). METHODS: Sixty-one patients with acute leukemia of initiatory treating were randomly divided into two groups, the 31 patients in the treated group were treated with SFI plus chemotherapy and the 30 patients in the control group treated with chemotherapy alone. The remission rate, changes of absolute number of peripheral mature neutrophils, T-lymphocyte subsets, serum levels of IL-6 and TNF-alpha before and after treatment were observed. RESULTS: The remission rate was higher in the treated group than that in the control group but the difference was insignificant (P > 0.05). The restoring of peripheral mature neutrophils in the treated group was higher than that in the control group, from the 3rd week of treatment, the difference was significant (all P < 0.05). CD4 and CD4/CD8 ratio after treatment increased in both groups, the increment was more obvious in the treated group than that in the control group significantly (P < 0.05). Serum levels of IL-6 and TNF-alpha lowered in both groups after treatment significantly(all P < 0.01), but the decrement was greater in the treated group with significant difference to the control group (P < 0.05). CONCLUSION: SFI could improve and regulate the immune function in acute leukemia patients undergoing chemotherapy and enhance the therapeutic effect. PMID- 12764906 TI - [Study on correlation between bone mineral density and syndrome type of TCM in patients with chronic obstructive pulmonary disease]. AB - OBJECTIVE: To explore the correlation between bone mineral density and Syndrome type of TCM in patients with chronic obstructive pulmonary disease (COPD) for providing the base of clinical integrative traditional Chinese and western medical therapy for the disease through the Syndrome typing and determination of changes in bone metabolism and bone density. METHODS: Bone mineral density (BMD) of lumbar vertebrae 2-4, femoral neck, Ward's triangle and trochanter in 27 COPD male patients, 25 male control subjects and 25 healthy persons were determined using dual energy X-ray absorptiometry, patient's Syndrome type, their blood levels of total protein, albumin, alkaline phosphatase, bone glaprotein, hydroxyproline, calcium, phosphate, urine levels of calcium/creatine and phosphorous/creatine as well as arterial blood gas were also determined. RESULTS: The BMD in COPD patients accompanied with respiratory failure or with course > 10 years was higher than that in COPD patients without respiratory failure or with course < or = 10 years, BMD in COPD patients of Fei-Pi-Shen type was lower than that in those of Fei-Pi, but the urine hydroxyprdine in the former was higher than that in the latter (all P < 0.05). CONCLUSION: (1) COPD is a risk factor for osteoporosis; (2) Shen Deficiency is the key and nucleus of secondary osteoporosis to COPD; (3) It is inferred that early regulation of Shen may be facilitated to prevent osteoporosis in COPD patients. PMID- 12764907 TI - [Clinical observation on treatment of multiple bone metastatic tumor of mammary cancer by combination therapy of 89Sr and Chinese herbal medicine]. AB - OBJECTIVE: To compare the clinical curative effects between 89Sr and its combination with the Guliu recipe (GLR, a Chinese herbal medicine) in treating multiple bone metastatic tumor of mammary cancer (MBM-MC). METHODS: By adopting the random sampling and grouping method, 89Sr alone (Sr) and 89Sr combined with CHM (Sr-GLR) were used in treating 86 and 40 patients with MBM-MC respectively. The efficacy of therapy were appraised according to the degree of ostalgia relieving and quality of life (QOF) in patients, and the effect of treatment on focal bone metabolism and bone marrow hematopoietic function were compared. RESULTS: The effective rate of Sr and Sr-GLR in relieving ostalgia was 83.72% and 95.00%, respectively (P > 0.05), the QOF improving and stabilizing rate of them 80.23% and 95.00% (P < 0.05), the effective rate on focal bone metabolism 59.30% and 52.50% (P > 0.05) and their hemo-toxicity 28.00% and 30.00% (P > 0.05). CONCLUSION: Sr-GLR is a combination therapy in treating MBM-MC with good effect, it could raise the patient's QOF, enhance the ostalgia relieving effect without increasing the hemo-toxicity of treatment. PMID- 12764908 TI - [Effect of tetramethylpyrazine on endothelin, von Willebrand factor and thromboxane A2 during cardiopulmonary bypass in patients of congenital heart disease with pulmonary hypertension]. AB - OBJECTIVE: To study the effect of tetramethylpyrazine (TMP) on the vascular endothelial cell (VEC) related humoral factors, including endothelin (ET), factor VIII related antigen (i.e. von Willebrand factor, vWF) and thromboxane A2(TXA2) in patients of congenital heart disease with pulmonary hypertension (CHD-PH) during cardiopulmonary bypass (CPB), and explore the clinical physiopathologic significance of them. METHODS: Thirty non-cyanotic patients of CHD-PH were randomly divided into the control group and the treated group. TMP was given to the treated group by intravenous dripping 3 mg/kg after anesthesia induction and adding 1 mg/kg in oxygenator during CPB. Blood samples were collected from radial artery at the time points of after anesthesia induction, 15 min after beginning CPB, 5 min after opening aorta, 20 min, 6 hrs and 24 hrs after stopping CPB, to determine the plasma contents of ET and vWF, as well as TXB2, the stable metabolite of TXA2. The pulmonary vascular reactivity 6 hrs (6h-PVR) after CPB and the mechanical ventilatory support time (VST) after operation were calculated. RESULTS: Levels of ET, vWF and TXB2 increased obviously during CPB, but the degree of increasing in the treated group was lower than that in the control group (P < 0.05), and the 6h-PVR and VST in the former were also lower than those in the latter respectively. CONCLUSION: TMP could obviously reduce the production of ET, vWF and TXB2 during CPB and relieve the pulmonary vascular reactivity after operation, indicating that TMP could reduce the injury of CPB on VEC, and is benefit to enhance the efficacy of treatment. PMID- 12764909 TI - [Regulatory effect of yishou tiaozhi tablet on lipids in patients with primary hyperlipidemia]. AB - OBJECTIVE: To study the regulatory effect of Yishou Tiaozhi Tablet (YSTZT) on blood lipids in patients with primary hyperlipidemia. METHODS: One hundred and sixty-eight patients with primary hyperlipidemia were randomly divided into two groups. The treated group was treated with YSTZT 4 tablets 3 times a day, and the control group was treated with Zocor by oral taking 1 tablet before sleep every evening. The therapeutic course for both groups was 60 days. The effect in regulating blood lipids in patients was observed. RESULTS: The levels of total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C), arteriosclerosis index (AI), apo-lipoprotein B (apoB), TC/HDL-C lowered and apoA/apoB ratio increased obviously in the treated group. As compared the effects between the two groups, the lowering of TC, AI and apoB in the two groups were similar (P > 0.05), effect of YSTZT in lowering TG was superior but in lowering of LDL-C was inferior to those of Zocor respectively (all P < 0.01). The effect of both remedies in elevating HDL-C was not satisfactory. The total effective rate of YSTZT was 91.9%, its individual effects were similar to those of SJZ respectively except in lowering TG and raising HDL-C. The adverse reaction occurred in YSTZT treatment course was mild without any influence on the medication. CONCLUSION: YSTZT had definite regulatory effect on the blood lipids and lipoproteins in patients with hyperlipidemia. PMID- 12764910 TI - [Influence of large amount of shengmai injection on blood coagulation in patients with chronic heart failure]. AB - OBJECTIVE: To investigate the effect of Shengmai Injection (SMI) on blood coagulation in patients with chronic heart failure (CHF). METHODS: Sixty patients with CHF were randomly divided into two groups, the 30 patients in the treated group were treated with SMI plus conventional treatment of western medicine, and the 30 in the control group treated with conventional treatment alone. The changes of cardiac function were observed and levels of plasma P-selectin, von Willebrand's factor (vWF) and D-dimer were determined. RESULTS: The total effective rate and the markedly effective rate in the treated group were higher than those in the control group respectively. The levels of P-selectin, vWF and D dimer lowered in both groups significantly after treatment, but the effect of lowering was better in the treated group than that in the control group. CONCLUSION: Hypercoagulative state exist in patients with chronic heart failure. SMI could improve the state in patients, which may reduce the occurrence and developing of emboic events to certain extent. PMID- 12764911 TI - [Clinical observation on treatment of Russula subnigricans poisoning patients by Ganoderma lucidum decoction]. AB - OBJECTIVE: To observe the effect of Ganoderma lucidum decoction in treating Russula subnigricans poisoning (RSP) patients. METHODS: The 14 patients of RSP in the treated group were treated with GLD (GLD, one dose was prepared by 100 g of Ganoderma lucidum decocted with water to 600 ml), on the base of conventional treatment, and 11 patients received conventional therapy in the previous year were taken as control. The clinical efficacy and parameters in them were compared, including the urine N-acetyl-D-glucosaminidase (NAG, which reflects the injury of kidney), the red blood cell and protein in urine, the alanine transaminase (ALT, which reflects the injury of liver), and the aspartate aminotransferase (AST, which reflects the injury of heart). RESULTS: A better clinical cure-markedly improving rate was showed in the treated group as compared with the control group, P < 0.01. In the treated group, red blood cell in urine disappeared after 24 hrs treatment in the majority of patients, urinary protein reduced obviously and the other three parameters reached the peak at the 3rd day then lowered gradually. In the control group, all the parameters increased continuously. Comparison between the parameters at corresponding time in the two groups showed significant difference (P < 0.01), those in the treated group were markedly lower than those in the control group respectively. CONCLUSION: GLD has good effect in treating RSP, could obviously lower the fatat rate of RSP. PMID- 12764912 TI - [Effect of congsheng capsule on intracellular calcium concentration in mice after ischemic cerebral injury]. AB - OBJECTIVE: To investigate the effect of Congsheng Capsule (CSC) in intracellular Ca2+ concentration in cerebral intra-synaptosomes of ischemia/reperfusion mice and analogous ischemic injured neurons to estimate the mechanism of CSC in antagonizing ischemic cerebral injury. METHODS: The concentration of Ca2+ in cerebral intra-synaptosomes of mice and in neurocyte suspension of fetal mice were determined using Fura-2/AM double wavelength fluorescence spectrophotometer. RESULTS: CSC 3 g/kg and 9 g/kg through gastrogavage could relieve the degree of calcium overloading of intra-synaptosomes in mice after cerebral ischemia/reperfusion. The CSC containing serum could inhibit the elevation of Ca2+ content in neurocytes induced by 1 hour hypoxia and glutamate of high concentration (200 mumol/L). CONCLUSION: CSC plays a role in antagonizing ischemic injury through relieving the calcium overloading after cerebral ischemia. PMID- 12764913 TI - [Effects of nourishing shen-yin recipe and warming recuperating shen-yang recipe on interleukin-1 and interleukin-6 activity in ovariectomy induced osteoporosis rats]. AB - OBJECTIVE: To compare the effects of the recipes for nourishing Shen-yin (NSY) and for warming recuperating Shen-yang (WRSY) on the activity of interleukin-1 (IL-1) and interleukin-6 (IL-6) in ovariectomy-induced osteoporosis rats. METHODS: Fifty-five female rats were randomly divided into five groups: the control, the sham operation, the ovariectomy, the ovariectomy plus NSY and the ovariectomy plus WRSY group. The medication was starting from one week after operation and lasted continuously for 3 months. The proximal tibia of rats were analyzed with bone histomorphometry, the activity of IL-1 was detected with mouse thymocyte assay and the activity of IL-6 was detected with IL-6 dependence cell proliferation assay. RESULTS: Three months after ovariectomy, the percentage of trabeculae bone volume (TBV%) of tibia in rats was markedly reduced, but the percentage of trabeculae absorption surface (TRS%) and trabeculae formation surface (TFS%) significantly increased, and the activity of IL-1 and IL-6 increased obviously. These abnormal changes could be reversed to different degree by NSY or WRSY, but the effect of the latter was superior to that of the former. CONCLUSION: The inhibitory effect of WRSY on IL-1 and IL-6 activity was significantly better than that of NSY, which is one of the mechanisms of its superiority in preventing osteoporosis induced by ovariectomy in rats. PMID- 12764915 TI - [Experimental study on effect of Astragalus extractum on canine isolated kidney during hypothermia perfusion and preservation]. AB - OBJECTIVE: To investigate the effect of Astragalus Extractum on canine isolated kidney during hypothermia perfusion and preservation. METHODS: Isolated kidneys in the control group were hypothermia perfused and preserved using conventional hypertonic adenine citrate solution (HC-A), and for those in the experimental group, using HC-A plus Astragalus extract instead. The changes of renal tissue construction were observed with light microscopy and electron microscopy. Moreover, the kidney transplantation model of dog was established to determine the changes of biochemical parameters before and after transplantation. Data were analysed synthetically. RESULTS: The ultrastructural injury in preserved kidney of the experimental group was significantly milder than that in the control group. Parameters determined in the early stage of transplantation showed that the blood creatinine level was significantly lower and the endogenous creatinine clearing value was higher in the experimental group than that in the control group, the difference was significant (P < 0.05). CONCLUSION: When Astragalus extractum is used in preserving kidney with hypothermia perfusion, it shows definite protective effect on the ischemic reperfusion injured kidney. PMID- 12764914 TI - [Effect of tonifying shen recipe on advanced glycosylation end products in aorta and serum lipids in ovariectomized rats]. AB - OBJECTIVE: To investigate the effect of tonifying Shen recipe (TSR) on levels of advanced glycosylation end products (AGEs) in aorta, serum lipids and lipid peroxidation in ovariectomized rats. METHODS: Rats were randomly divided into the sham group, the ovariectomized group and the TSR group, in which the rats were treated with TSR for 13 weeks starting from 2 weeks after ovariectomy. Blood sample was taken out from rat at the end of the experiment after 24 hrs fasting for determination of lipids and lipid peroxidation, and the animal was sacrificed, the aorta was taken out for detecting AGEs. RESULTS: No significant difference was found between groups in levels of total cholesterol and low density lipoprotein cholesterol. In comparing with the sham group, levels of aortic AGEs, serum triglyceride (TG), oxidized low density lipoprotein (OX-LDL) and malondialdehyde (MDA) in the ovariectomized group were obviously higher (P < 0.05 or P < 0.01), and levels of high density lipoprotein cholesterol (HDL-C), apo-lipoprotein A-I (apoA-I) and activity of superoxide dismutase (SOD) were lower (all P < 0.01). While in the TSR group, as compared with the ovariectomized group, the above-mentioned abnormal changes, excepting for TG, were all reversed to certain degree (P < 0.05 or P < 0.01). CONCLUSION: TSR displays its cardiovascular protecting effect in ovariectomized rats through lowering the AGEs content in aorta, reducing the serum levels of OX-LDL and MDA, raising the levels of serum HDL-C and apoA-I and increasing SOD activity. PMID- 12764916 TI - [Effect of Tripterygium glycosides on thyroid function and auto-antibody in patients with Graves disease]. PMID- 12764918 TI - [Clinical observation on treatment of 48 patients with systemic lupus erythematosus using fuzheng jiedu recipe]. PMID- 12764917 TI - [Clinical observation on treatment of traumatic intracranial hematoma by Flos carthami combined with radix Acanthopanacis senticosi injection]. PMID- 12764919 TI - [Longevity and cancer]. PMID- 12764920 TI - [Exploration on the feasibility of applying TCM yin-yang theory in genome study]. PMID- 12764921 TI - [Scientific technological retrieval of TCM and its commission]. PMID- 12764922 TI - [Progress on study of experimental physico-chemical indexes related with ischemic stroke]. PMID- 12764923 TI - [The 6th National Conference on Hematology of Ingrative Traditional Chinese and Western Medicine]. PMID- 12764924 TI - A pound of flesh. PMID- 12764925 TI - Caught off guard. SARS reveals gaps in global disease defense. PMID- 12764926 TI - Acting locally. In curbing greenhouse gas emissions, states go it alone. PMID- 12764927 TI - Hybrids take off. Engineers reconsider cross-bred propulsion. PMID- 12764928 TI - Boxed out. Science loses as the U.S. tightens visa rules. PMID- 12764929 TI - Law and disorder. A quantum steam engine gets around the second law. PMID- 12764930 TI - Trade globalization. It is nearly two centuries old and likely to continue. PMID- 12764931 TI - The Abyss transit system. James Cameron commissions the making of robots for a return to the Titanic. PMID- 12764932 TI - Sign here. Will a scientist need a legal opinion before starting the next experiment? PMID- 12764933 TI - Codified claptrap. The Bible Code is numerological nonsense masquerading as science. PMID- 12764934 TI - One last look. PMID- 12764935 TI - Shoot this deer. PMID- 12764936 TI - The unearthly landscapes of Mars. The Red Planet is no dead planet. PMID- 12764937 TI - Self-repairing computers. PMID- 12764938 TI - Pandora's baby. PMID- 12764939 TI - The dawn of physics beyond the standard model. PMID- 12764941 TI - Cochlear implants. To hear again. PMID- 12764940 TI - Chain letters & evolutionary histories. PMID- 12764942 TI - Why do hangovers occur? PMID- 12764943 TI - A monster problem meets its match when the Monster Cigarette comes to town! PMID- 12764944 TI - Failure to claim your overtime. It can hurt your patients, your profession, and yourself. PMID- 12764946 TI - [Polyethylene disease]. AB - PURPOSE OF THE STUDY: The experience obtained during revision surgery and findings of polyethylene granulomas in surrounding tissues of replacement as well as marked differences in the viability of implants resulted in the study of polyethylene disease and its basic mechanisms producing the development of osteoaggressive granulomas. We investigated the morphology of particles and their number in tissues surrounding the implant. The aim of our study was to develop a method for the detection of polyethylene particles in tissues, to identify different types of wear and to assess factors that may influence the viability of joint arthroplasty in general. MATERIAL: Every revizion of joint arthroplasty performed during the last five years was evaluated in terms of the presence of polyethylene granules and the viability state of articular polyethylene inserts. A total of 55 samples were taken from tissues around loosened endoprostheses. The location of each sample was exactly determined. METHOD: A technique was developed to identify wear particles and to visualize them after all organic structures of a polyethylene granuloma were dissolved with nitrogenic acid. RESULTS: The viability of articular polyethylene implants showed extreme differences in relation to different periods of manufacture and probably also to different methods of sterilization. Articular inserts sterilized with formaldehyde (the method used at the beginning of arthroplasty in our country) showed the highest viability and the lowest wear. The polyethylene particles present in tissues surrounding the implant were characterized in terms of morphology and size. DISCUSSION: The comparison of literature data and our results has revealed that there are many unknown facts about the quality and structure of polyethylene. The method of sterilization also appears to play a role. Because the issue is complex, we were not able to identify all factors leading, in some cases, to an early and unexpected failure of the implant and we consider further investigation to be necessary. CONCLUSIONS: Polyethylene disease is an important factor limiting the viability of joint arthroplasty. It results from a complex interaction of polyethylene particles arising by wear with surrounding tissues. The particles, less than 0.5 micron in size, are phagocytized by macrophages and, by complex mechanism of expression of inflammation mediators, they result in the inhibition of osteogenesis and activation of osteoclastic processes. The previous methods of sterilization with formaldehyde vapors apparently reduced wear influenced the resistance of polyethylene to wear to a lesser degree. A method was developed to detect these particles and to characterize their morphology in the tissues of a polyethylene granuloma. PMID- 12764947 TI - [Two-stage reimplantation using spacers--the method of choice in treatment of hip joint prosthesis-related infections. Comparison with methods used from 1979 to 1998]. AB - PURPOSE OF THE STUDY: Several therapies are available for the treatment of deep infection in total hip arthroplasty but none is completely successful; there is no consensus on an optimal method. The aim of this study was to evaluate the treatment used in our institution and its outcomes over the last 20 years. In each method, the success of treatment was evaluated in terms of both infection control and restoration of function in the treated joint. MATERIAL: A total of 172 patients with infected total hip replacements were treated at the First Orthopedic Clinic of the First Faculty of Medicine, Charles University, Prague, between 1979 and 1998. Our sample consisted of 132 patients, 92 men and 40 women. Resection arthroplasty was performed in 62 patients. Two-stage reimplantation was used in 64 patients. Two-stage reimplantation involving skeletal traction was applied in 35 and a block spacer was used in 29 patients. The remaining patients were treated by other techniques. METHODS: The type of infection was classified according to the Coventry system. The outcome of surgery was assessed on the basis of the Tsukayma rating system, radiographic findings and the Harris hip score. RESULTS: The average follow-up time from the definitive operation was 70.8 months. In the patients who had resection arthroplasty only, the cure rate of infection was 91.9%. However, an increase in the Harris hip score, as compared with the condition before surgery, was low (9.7 points). In the patients treated by the two-stage reimplantation without a spacer but with skeletal traction, the cure rate of infection was 94.3% and the Harris score increased by 20 points. The patients who were treated by two-stage reimplantation with a spacer showed an infection cure rate of 96.5% and an increase in the Harris score by 29 points. This increase was higher by 9 points in comparison with the patients who had reimplantation without the use of a spacer. An even greater difference (28.2 points) was found when the outcomes of this technique were compared with those of resection arthroplasty. The incidence of spacer dislocation in 21% of the cases was an unexpected finding. DISCUSSION: No great differences in outcome in terms of infection cure rate were found among the methods used, i.e., two-stage reimplantation facilitated a better function for the hip joint than Girdlestone's operation. The use of a spacer in two-stage reimplantation ensured a greater comfort for the patient during treatment and gave better results in terms of joint function than treatment without a spacer. The use of a cemented spacer is an optimal method that not only ensures the stability of a limb during the period necessary for infection control but also provides conditions for the prospective implantation of a new prosthesis. The spacer also permits delivery of high-dose local antibiotics released from the cement as well as makes space for a long-term application of antibiotic-containing lavage. CONCLUSIONS: The rate of success in the treatment of an infected hip arthroplasty and the possibility of preserving the implant and thus enabling the patient to move comfortably are currently high. The prerequisite is early diagnosis and a radical surgical approach that involves the use of a method leading to the most effective eradication of infection and the maintenance of a good function for the joint. The reimplantation of a new prosthesis, after removal of the previous one and debridement of all infected tissue and material, combined with a targeted antibiotic therapy, is the method of choice for both the patient and the surgeon. PMID- 12764948 TI - [Restoration of elbow joint flexion by transfer of the pectoralis major muscle in patients with arthrogryposis multiplex congenita. Part II. Results of electromyographic and histologic examinations]. AB - PURPOSE OF THE STUDY: In the framework of a prospective study on transposition of the m. pectoralis major according to Clark in patients with type I arthrogryposis multiplex congenita, electromyography was carried out in order to determine pre operative states of the elbow joint flexors and m. pectoralis major and then the post-operative electric activity of a transposed muscle and to correlate changes with clinical findings. Histological examination was performed to reveal changes in muscle morphology and to complete a comprehensive assessment of muscle transposition. MATERIAL AND METHODS: Electromyography was carried out on nine upper limbs of five pediatric patients aged 4.3 to 8.9 years. Using a needle electrode, activities of the elbow flexors (m. biceps and m. brachialis), m. pectoralis major, m. triceps brachii and m. deltoideus were examined. In the post operative period, activity was repeatedly measured in both the transposed and non transposed parts of the m. pectoralis major. In one patient, histological examination of muscle tissue was performed at 26 months after transposition; light microscopy of paraffin-mounted sections stained with hematoxylin-eosin was used. RESULTS: Out of seven arms examined by electromyography before muscle transfer, six showed complete and one incomplete atrophy of the m. biceps brachii and m. brachialis. The m. pectoralis major had a five- to four-degree electric activity, which provided enough strength for transposition. Post-operative examination revealed changes leading to re-innervation of the transposed muscle, which corresponded to a partial denervation of the muscle followed by repair of innervation. None of the muscles was markedly atrophic due to denervation. In muscles with a higher electric activity, clinical outcomes were better, although electric activity always slightly exceeded clinical activity. In terms of electric activity, the transposed muscle was stabilized a year after surgery. Non transposed parts of the muscle were not damaged by the surgical procedure, as shown by electromyography. Histological examination showed the muscle at a state of partial atrophy but with signs of ongoing regeneration of muscle fibers. DISCUSSION: No data on examination of the electric activity of the m. pectoralis major following its transposition in patients with arthrogryposis multiplex congenita have been reported in the literature. Electromyography in this study proved useful for providing information on the electric activity of a muscle before transposition and on contractility of the muscle after surgery; it also allowed us to distinguish between a mechanical failure of transfer and muscle atrophy due to neurogenic or vascular causes. All transposed muscles that were examined revealed changes indicating a minimum denervation followed by re innervation. This finding was confirmed by the results of histological examination. CONCLUSIONS: Electromyography showed that the electric activity of a transposed muscle corresponded to the clinical presentation of this muscle and thus became an indispensable part of both pre- and post-operative examination. Both electromyographic and histological examination confirmed the applicability of the treatment described here. PMID- 12764949 TI - [Osteosynthesis of proximal femoral fractures using short proximal femoral nails]. AB - PURPOSE OF THE STUDY: A group of 147 patients with proximal femoral fractures treated with the use of the proximal femoral nail, manufactured by the Synthes company, in the period between October 1997 and June 2001 were retrospectively evaluated. MATERIAL: The group of 147 patients whose average age was 69 years consisted of 59 men (average age, 59 years) and 88 women (average age, 75 years). The most frequent fracture occurring in 54% of the patients was an unstable pertrochanteric fracture (Kyle III); persubtrochanteric and subtrochanteric fractures were treated in 27% and 19% of the patients, respectively. METHODS: In 1997, a new implant, the proximal femoral nail manufactured by the Synthes company (PFN), was introduced in the Czech Republic. In addition to all advantages of a nail to be implanted intramedullary, it has several other favorable characteristics: its length is 240 mm, pre-drillings is not necessary, it can be dynamically locked, it has a high rotation stability, and mechanical stress concentration on the implant-bone interface is low. This evaluation is based on the type of fracture, the duration of healing, the duration of surgery and X-ray exposure. RESULTS: The patients were followed up at 6 months, 3, 6 and 12 months, and once a year. The fracture healed in the anatomical position by 6 months in 95% of them. The average operative time was 56 min and X-ray exposure lasted on average 1 min. We recorded 14 intra-operative complications in nine patients, which included incomplete reduction in four cases, fixation in distraction in two, incorrect length of screws in one, fracture at the site of distal locking in two and incorrect insertion of femoral neck screws in five cases. Early post-operative complication involved seven cases of hematoma in the operation wound with the necessity to re-operate in five cases. Late complications occurred in two patients and included pseudoarthrosis and necrosis of the femoral head after healing. Most of the complications were found in subtrochanteric fractures. DISCUSSION: The treatment of unstable fractures of the proximal femur is still associated with some failures. The reasons are: disregard for biomechanics, overestimation of the potentials of new surgical techniques or new implants or poor adherence to established procedures. PFN is a novel, modern implant based on experience with the gamma nail. Since relevant literature data are very few, any comparison of our results is very difficult. The lower average age of our group was due to the intentional selection of our patients for this type of surgery. CONCLUSIONS: PFN is an excellent implant for the treatment of unstable fractures of the proximal femur. The terms of successful outcome include a good understanding of fracture biomechanics, correct indication and exactly performed osteosynthesis. PMID- 12764950 TI - [Initial experience with implantation of the cemented Beznoska/S.V.L. total knee joint endoprosthesis]. AB - PURPOSE OF THE STUDY: The authors present the results of their first trial of a new, Beznoska/S. V. L. type, knee prosthesis in order to introduce it to a broad orthopedic public. MATERIAL: Clinical and radiological evaluation was carried out on 34 knee prostheses implanted in 31 patients between September 1997 and October 1999. The average patient age at the time of surgery was 71.3 years and the average interval between surgery and assessment was 22.4 months (range 6 to 31 months). All patients underwent implantation due to primary or secondary gonarthosis. A brief description of the implant and the instrumentation and used surgical technique is provided. METHODS: Clinical outcomes were evaluated according to the "Knee-Society Clinical Rating System" by John N. Insall. The system classifies both knee joint parameters and knee function. X-ray films were assessed on the basis of the "Knee-Society Total Knee Arthroplasty Roentgenographic Evaluation and Scoring System" by Frederic C. Ewald. In standardized X-ray projections, this allowed us to evaluate the implant position as well as radiolucent lines. RESULTS: On stability evaluation, we found anteroposterior instability up to 5 mm in 85% of the implants and mediolateral instability up to 9 degrees in 97% of them. A maximum flexion of 90 degrees to 120 degrees was achieved in 91% of the implants. The final outcome in terms of knee score was on average 80.3 points (range, 40 to 97 points), which was a very good result. Evaluation by function score showed that only 17% of the patients were not able to walk farther than 500 meters. Although 82% of them had to hold on a railing when going upstairs, all were able of stair ascent and descent; 13% had to use a walking stick permanently. The average function score was 68.4 points (range, 30 to 100), which was a good outcome. Radiograms in anteroposterior projection, assessed according to Ewald, showed the average femoral flexion angle (alpha) to be 95.2 degrees, the average tibial angle (beta) to be 89 degrees and the total valgus angle (omega) to be 3.2 degrees. In lateral projection, the femoral flexion angle (gamma) was on average 2.5 degrees and the average tibial angle (delta) was 86.7 degrees. An optimal position of the patella was achieved in 27 implants. Five radiolucent lines, up to 1 mm, were found in zone 1 of the femoral component and further lines were observed in zones 1, 2 and 4 of the tibial component. DISCUSSION: When assessing the results by the knee and function scores, it had to be taken into consideration that the average age of the patients was 71.3 years. At this age, walking without a stick, or stair ascent or descent without the use of a railing can hardly be expected. The average result of 68.4 points achieved can, therefore, be considered a very satisfactory outcome. The values shown by X-ray examination were close to the normal condition. The patients were also asked for their subjective opinion of the effect of arthroplasty; 97% of them regarded the effect as good or very good, only one patient reported no benefit. CONCLUSIONS: The results of the first trial of the use of a cemented prosthesis, type Beznoska/S. V. L., in total knee arthroplasty are presented, together with practical recommendations. Although the patient sample was small and the follow-up period short, the results are promising and suggest excellent prospects for this implant. PMID- 12764952 TI - [Current views on diagnosis of osteoarticular tuberculosis]. AB - This paper presents current methods used in the diagnosis of osteoarticular tuberculosis in the Czech Republic. It draws attention to difficulties and errors associated with the diagnosis of this disease. The aim of this study is to provide a comprehensive review of relevant diagnostic methods, from medical history to imaging techniques. A misdiagnosis and/or a late establishment of the diagnosis, which may result in inadequate primary therapy, prolong both the time needed for complete treatment and the duration of absenteeism. These may eventually have negative economic and social implications. PMID- 12764951 TI - [Case report of extensive metallosis in extra-articular tissues after unicompartmental knee joint replacement]. AB - The authors present a case of extensive metallosis of the knee joint resulting from the secondary abrasive wear of components of a unicompartmental alloplasty. Eight years after medial condyle replacement, an eighty-year-old patient presented with a break of the femoral component. This resulted in a rapid wear of tibial polyethylene and the development of secondary friction between two metal surfaces. The generation of a large number of metal particles produced an increase in the actual metal surface. This extensive articular metallosis affected not only intra-articular but also extraarticular tissues and the subchondral bone. Histological examination confirmed the presence of a large amount of opaque pigment in histiocytes. The condition was treated by total knee arthroplasty with resection of the posterior cruciate ligament (stabilizing plateau). The authors discuss the causes of metallosis development and its various types; they draw attention to the scarcity of literature data on this condition. Metallosis is a combined chemical and toxic reaction which, if the contact surface of a metal implant is large, may cause extensive damage to the surrounding tissue, the bone included. PMID- 12764953 TI - Nurses have a key role in reducing STI levels. PMID- 12764954 TI - Setting priorities for local services. PMID- 12764955 TI - Viral sexually transmitted infections. PMID- 12764956 TI - Reducing the transmission of sexually transmitted infections. AB - The incidence of sexually transmitted infections (STIs) diagnosed in UK genitourinary medicine clinics has increased significantly over the past decade. Nurses are becoming more involved in developing and delivering interventions that aim to control the spread of STIs. A number of control strategies for STIs used by clinicians are reviewed in this paper. PMID- 12764957 TI - Adverse health-care events: Part 1. The nature of the problem. AB - Adverse events are a significant cause of unnecessary harm in health care and can lead to both physical and psychological injury and, in some cases, death. This paper, the first in a series of four, outlines the nature and extent of the problem. The overall aim of the series is to enhance knowledge levels among nurses in an attempt to reduce the number of adverse events. PMID- 12764958 TI - Factors that impact on the infection control capability of gloves. AB - The use of gloves can reduce the risk of cross-infection for patients and health care professionals. This is dependent, however, on the correct type of glove being selected for the appropriate job. Care must be taken to ensure that gloves are stored and worn correctly, to reduce the chance of exposing them to factors that may damage their integrity. PMID- 12764959 TI - A nurse-led randomised trial of pressure-relieving support surfaces. AB - A nurse-led trial is currently collecting data comparing interventions for the prevention and treatment of pressure ulcers. The aim is to provide reliable guidance on the relative merits of alternating-pressure mattresses and overlays for people at moderate to high risk of pressure ulceration. This paper outlines the main objectives, methodology and progress of the study. PMID- 12764960 TI - Developing a framework to assess competence in leg ulcer care. AB - To provide effective and safe patient care, it is imperative for all practitioners to possess a core set of skills. This paper describes the development of a framework for assessment of competence in carrying out Doppler assessments and compression bandaging for practitioners involved in caring for patients with leg ulcers in one institute of higher education. PMID- 12764961 TI - Providing breastfeeding support to ethnically diverse groups of mothers. AB - To encourage more mothers to breastfed a breastfeeding reassurance and guidance group was set up to provide professional and peer support and advice for new mothers. Few mothers from ethnic minorities attended, which prompted research into women's views of local services. Problems in accessing services by public transport and safety issues were raised. PMID- 12764962 TI - Good practice for keeping stroke patients and carers informed. AB - Stroke patients and their carers can have many questions about the mental and physical effects of the condition, hospital procedures and treatments, and prospects for the future. This paper discusses issues involved in giving information to patients--some of whom may be experiencing serious communication problems--as well as those looking after them, and offers solutions. PMID- 12764963 TI - Safeguarding children: 4. Needs of refugees and asylum seekers. AB - The problems faced by all refugees and asylum seekers arriving in Britain are great but unaccompained children have a particularly difficult time, separated from the people and places they know, and experiencing difficulties with language and a strange environment. This fourth paper in our series on child protection examines how health- and social-care staff can help. PMID- 12764964 TI - Achieving lifelong learning for all health service professionals. AB - Plans are in place to make lifelong learning accessible for all health-care staff, whatever their profession. However, as the second of two articles on continuing professional development reveals, there is still wide variation across trusts in access to education. PMID- 12764965 TI - Home guards. PMID- 12764966 TI - Take the danger out of working. PMID- 12764967 TI - NICE try. PMID- 12764968 TI - 'It's still hands-on'. Interview by Laurence Dopson. PMID- 12764969 TI - Many happy returners. PMID- 12764970 TI - Laughing through the pain. PMID- 12764971 TI - The eye in the door. PMID- 12764972 TI - One step at a time. PMID- 12764973 TI - Using self-efficacy as a client-centred outcome measure. AB - BACKGROUND: The concept of self-efficacy has been proposed as a suitable outcome measure in psychiatry. Self-efficacy refers to an individual's belief that he or she can control important aspects of his or her life. An audit of 67 consecutive clients attending an acute psychiatric day unit was conducted, measuring self efficacy at admission to the unit and at discharge. CONCLUSION: The concept of self-efficacy was easily explained to clients, and was measured using a 15-point questionnaire. Self-efficacy increased following treatment in the day unit, and this increase appears to reflect an underlying change in the individual's self belief. PMID- 12764974 TI - Work-based learning: making a difference in practice. AB - Nurses play an increasingly crucial role in ensuring that patients receive the best possible care, and strive to lead innovations in health care. Changing practice is not easy and many nurses do not have the leadership skills or confidence to push for change. Therefore, they need to know that they are supported and encouraged to bring about change in nursing practice. Primary care trusts in the west of Berkshire and a university based in Reading have worked together to respond to this challenge by developing and implementing a flexible, accredited, work-based educational programme. The programme ensures that patients remain at the heart of learning by enabling nurses to lead and influence practice. PMID- 12764975 TI - Subcutaneous insulin injection technique. AB - Insulin injection technique has far greater bearing on glycaemic control than is often recognised. This article discusses the optimal needle length for different injection sites and highlights other crucial aspects, such as needle re-use, importance of adequate mixing of cloudy insulin, and the way pen devices should be stored. PMID- 12764977 TI - RN blows whistle on staffing: was there retaliatory termination? PMID- 12764976 TI - General prescribing principles. Errors and near misses. AB - Remember... Mistakes happen. Know the common medication errors. Take active steps to avoid them in your practice. Know the procedures for reporting errors and near misses. Reflect and learn from mistakes and near misses. Know about your local compliants procedures. PMID- 12764978 TI - 'Excited utterance' exception to hearsay rule. Case on point: Montanez v. State, 2003WL 1856529 S.W.3d--TX. PMID- 12764979 TI - KY: unlicensed nurse expert's testimony excluded: selecting 'wrong expert' can be fatal to case. PMID- 12764980 TI - MN: terminated nurse reported abuse of elderly: no retaliatory termination found- benefits denied. PMID- 12764981 TI - Nurse's testimony impacts on patient's 'capacity' to execute will. Case on point: Stephenson v. Stephenson, 2003 WL 1618090 S.W.3d--TN. PMID- 12764982 TI - New York privacy law. Unraveling the interplay between New York State law and federal requirements. Which takes precedent. PMID- 12764983 TI - Burning mouth syndrome. A retrospective analysis of clinical characteristics and treatment outcomes. AB - Burning mouth syndrome is a condition characterized by burning sensations of the oral cavity in the absence of physical abnormalities of the mucosa or a detectable underlying medical disorder. It is a multifactorial disorder with unclear etiology, affecting predominatly middle-aged women. Multiple approaches to treatment have been described in the literature, with few controlled clinical trials regarding their efficacy. The objectives of this retrospective study were to: 1. determine the epidemiologic characteristics of BMS patients referred to an oral medicine practice; 2. determine if BMS classification correlates with response to treatment; 3. determine the efficacy of a variety of known therapies for BMS. A database was constructed from the charts of 150 consecutive patients diagnosed with BMS; and these charts were reviewed. Patients were classified according to previously published criteria for BMS. Presumed etiologies were grouped into depression/anxiety-associated; hematinic deficiencies, including iron, folate and vitamin B complex; oral habits: and idiopathic BMS. Treatment approaches were divided into seven categories: soft desensitizing appliance; tricyclic antidepressants (TCA); benzodiazepines (BZD); topical analgesics; hematinic supplements; habit awareness counseling; and multi-modal therapy (combining two or more of the above). Improvement was recorded using a zero to 100% VAS scale and classified as no relief (0%); mild (0-40%); meaningful/moderate (41-80%); and profound relief (81-100%). Burning mouth syndrome without any identifiable cause (idiopathic) was diagnosed in 33 patients (46.6%). Patients were followed up at one month (4 weeks) after the initial visit. Nine patients (12.7%) reported profound relief; 17 patients (23.9%) reported meaningful relief; 39 patients (54.9%) reported mild relief. This retrospective review showed no significant correlation between classification of BMS and response to therapy. The most effective treatment modalities were habit awareness, followed by TCAs. PMID- 12764984 TI - Idiopathic bone cavity. A report of recurrent lesions and their management. AB - Idiopathic bone cavity is a lesion that has been described as a bony cavity lacking an epithelial lining. Other names have been used, which are not really descriptive of this process. We present a case that involved bilateral idiopathic bone cavities that were treated initially by surgical exploration. Both lesions recurred and were treated with a second procedure consisting of exploration and grafting with platelet--rich plasma and porous hydroxyapatite. The lesions have not recurred 14 months following the second surgery. PMID- 12764985 TI - Haemophilus influenza cellulitis. A review and case report. AB - Buccal cellulitis resulting from Haemophilus Influenzae type B (Hlb) is an uncommon yet potentially life-threatening illness that afflicts the facial soft tissues of the very young. Early recognition is essential for the effective treatment of this illness. A clinical case of Haemophilus Influenzae buccal cellulitis is presented, accompanied by a discussion of the presenting symptoms, diagnosis and treatment of this unusual childhood infection. PMID- 12764986 TI - [Early jejunal nutrition with combined pre- and probiotics in acute pancreatitis- prospective, randomized, double-blind investigations]. AB - BACKGROUND: Development of infection of pancreatic tissue in patients with severe acute pancreatitis dramatically increases morbidity and mortality. Colonisation of the lower gastrointestinal tract and oropharynx, mostly with gram-negative but sometimes also gram-positive bacteria is known to precede the contamination of the pancreatic tissue by a few days. A few specific lactic acid bacteria such as Lactobacillus plantarum 299 were effective in preventing colonisation of the gut by potential pathogens, to reduce endotoxemia and to stimulate the gut-associated lymphatic system (GALT) and the immune system. METHODS: Patients with acute pancreatitis, arriving within 48 hours after onset of disease showing typical clinical picture and laboratory signs of pancreatitis (plasma amylase > 200 U/l, CRP > 150 mg/l and an Imrie-score = or > 3) were randomised into two groups During the first week the treatment group received a freeze-dried preparation containing 10(9) live Lactobacillus plantarum 299 together with an oat fibre substrate. The control group received a similar preparation, but the Lactobacillus plantarum 299 had been inactivated by heat. For seven days the treatment was repeated twice every day. The preparations were delivered to the hospital in sachets so the content was unknown to the investigators, staff and patients. RESULTS: Forty five patients completed the study before there was indication that one group differed from the other in a statistically significant way, at which time the study was discontinued and the code broken. At this time 22 patients had received treatment with live and 23 patients with heat-killed Lactobacillus plantarum 299. Infected necrosis and abscesses occurred in 1/22 (4.5%) in the treatment group vs. 7/23 (30%) (p = 0.023) in the control group. The length of stay was 13.7 days in the treatment group vs. 21.4 days in the control group (not statistically significant). CONCLUSIONS: Supplementing Lactobacillus plantarum 299 is an effective tool to prevent pancreatic sepsis, to reduce the number of operations and length of stay. The only patient who developed sepsis in the treatment group did so eight days after the treatment had been discontinued. One week treatment, as in the present study, is too short. It should be provided for at least 2 weeks, or more appropriately, as long as the patients are treated with antibiotics or have signs of GI colonisation. PMID- 12764987 TI - [Radioguided excision of non-palpable breast lesions with simultaneous sentinel lymph node biopsy]. AB - As a result of mammographic screening nonpalpable breast lesions are found with increasing frequency. The surgical management of these lesions presents two main problems: (1) How to localize the lesion pre- and intraoperatively; (2) What should be the extension of the axillary dissection for malignant lesions. This study is aimed to evaluate the technical feasibility of radioguided excision of nonpalpable breast lesions and the possibility of performing simultaneous sentinel lymph node biopsy for malignant lesions. Radioguided breast excisions have been performed in our Department since November 2000. The basis of this method is that radioisotope labelled colloid is injected into the lesion under stereo tactic or sonographic guidance. A gamma-probe is used intraoperatively to locate the lesion and guide its excision. Dual agent guided technique is used for sentinel lymph node biopsy. Radioguided breast excision was performed on 85 patients (93 lesion) till February 2002. The localization and excision were successful in 84 patients (92 lesions) (99%). Multifocal lesions were successfully localized and excised in 7 patients. There were 61 parenchyma lesions (66%) and 31 microcalcifications (34%). Localization was performed under sonographic guidance in 44 patients (52%) and under stereotactic guidance in 40 patients (48%). There were 25 benign (27%) and 67 malignant (73%) lesions. Second surgical procedure (mastectomy) was needed in 4 patients (6.5%) because of histologically incomplete excision. Sentinel lymph node biopsy was performed in 50 patients. The biopsy was successful in 45 patients (90%). The sentinel lymph node was histologically positive in 2 patients (4.4%). Radioguided localization with the intraoperative use of a gamma-probe is an easy, rapid and highly accurate technique for removing nonpalpable breast lesions and allows simultaneous sentinel lymph node biopsy for malignant lesions. PMID- 12764988 TI - [Role of functional investigations in gastrointestinal surgery]. AB - The understanding, complex analysis and extensive knowledge about the physiology of the gastrointestinal tract are bases of functional and reconstructive surgery. We present our routine physiological tests. Several clinical syndromes may be related to motility disorders of the esophagus, stomach, colon and the anal sphincter apparatus. We show how functional tests influence surgery. Preoperative pH monitoring and esophageal manometry are essential in the planning of surgical treatment of GORD. The lower gastrointestinal functional tests help in the treatment of anal incontinence. Screening of the anal sphincter helps in the decision when to close non permanent stomas, and the inspection benefits haemorrhoidectomies on patients with poor anal sphincter status. We are present our initial practice with the functional tests. PMID- 12764989 TI - [Thoracic epidural anesthesia improves the gastric microcirculation during experimental gastric tube formation]. AB - We studied the effects of gastric tube formation and thoracic epidural anaesthesia (TEA) on gastric microcirculation, hemodynamic and bowel motility changes. The experiments were performed on pentobarbital-anesthetized, mongrel dogs. Mean arterial pressure (MAP), gastric and mesenteric blood flow (Transonic Systems Inc.) and small bowel motility changes (strain gauge technique) were monitored. The stomach was prepared according to the method of Akiyama. TEA was induced by injecting Bupivacain 1 mg/kg. Orthogonal polarization spectral imaging (OPS) technique (Cytoscan A/R, Cytometrics, PA, USA) was used to observe in vivo microcirculatory changes. The gastric pull-up caused significant decrease in intramucosal pH and red cell velocity (RBCV) in terminal arterioles of the upper part of gastric tube both on the mucosal and serosal side. After TEA the RBCV returned to baseline level and the intestinal motility index significantly increased. Functional capillary density of the mucosa or subserosa did not change during the experiments. MAP was significantly reduced by 30%, while the arterial blood flow in gastric and mesenteric arteries was significantly increased. TEA significantly improves microcirculation of the distal portion of the gastric tube and increases intestinal motility. These results show that epidural anaesthesia is favourable and should be recommended during reconstructive oesophageal surgery. PMID- 12764990 TI - [Role of procalcitonin rapid test in the differential diagnosis of uninfected and infected forms of acute pancreatitis]. AB - We analysed the clinical value of the procalcitonin quick test (PCT-Q; BRAHMS Diagnostica, GmbH, Berlin) in infected pancreatic necrosis verified by guided fine-needle aspiration (FNA). In a prospective, controlled study the results of 24 patients were evaluated during 2001. PCT-Q test was performed in patients with necrosis verified on CT scan and/or septic symptoms. If PCT-Q test was positive or septic complication (infected necrosis or abscess) developed CT or US guided fine-needle aspiration was performed with Gram staining and bacterial culture of the sample. Positive FNA result was indication for surgery with repeated staining and bacterial culture of the surgical specimen. Septic complications of pancreatic origin developed in 12 patients. Comparing the results fine-needle aspiration was more authentic with a sensitivity of 92% and a specificity of 100%, while the sensitivity of the PCT-Q test remained 75% and its specificity 83%. Comparing abscess and infected necrosis, significantly higher procalcitonin values were detected in patients with necrosis. These results show that PCT-Q test can be a possible non-invasive method besides fine-needle aspiration. Elevated levels of procalcitonin (higher than 2 ng/ml) clearly suggest infection of the necrosis, while lower values do not exclude the possibility of local septic progression. PMID- 12764991 TI - [The use of ROMICRO set in plastic surgery]. AB - More than thousand plastic operations were performed in our departments with ROMICRO-set during the last ten years. First we applied it for reconstructive and aesthetic augmentation of the breast. Thereafter, we developed new technique for gynecomastia and reconstruction of diastasis of the m. rectus abdominis sheath. According to our experience, the ROMICRO-set is a useful tool in plastic surgery. The ROMICRO-set can be used through a small incision to achieve three dimensional views with excellent exposure and lighting. PMID- 12764992 TI - [To the Editor in Chief: Left-sided gallbladder]. PMID- 12764993 TI - [Percutaneous portal vein embolization before major hepatic resection]. AB - Major liver resection cannot be performed when the remaining liver mass is too small. Preoperative embolization of the portal vein (PVE) helps to increase the volume of the non-tumorous liver segments, and patients' liver function will remain stable postoperatively. CT, MRI, CTAP examinations help to decide about surgery. Volume measurements are performed based on data of CT scans. PVE is indicated, when the remaining, non-tumorous liver volume is too small. The procedure starts with percutaneous portal vein catheterization, then selected portal vein branches are embolized with a mixture of contrast material, antibiotics and Gelfoam particles. The patients stay in the hospital 3-5 days after the procedure. Control CT-volumetry is done after 3-6 weeks. From November 2001 to April 2002 3 patients were selected to have this procedure. PVE of the right portal branches were performed successfully in two cases. Control CT volumetry showed significant increase of the volume of the left liver lobe. One patient underwent successful right hemihepatectomy. On the third patient we could not catheterize the portal vein. PVE is a relatively safe and tolerable procedure. The increased remaining liver volume helps in performing major liver resection. PMID- 12764994 TI - [Sapheno-peritoneal shunt (Routte-operation) in the treatment of refractory ascites]. AB - We describe the results of an operative method applied in patients suffering from refractory ascites. Instead of the well-known shunt-implants we started to re-use Ruotte's method, which was published 100 years ago. According to our observations the anastomosis formed by the long saphenous vein and the peritoneal surface eliminates not only the known complications of the traditional shunt-operations but also provides an effective early and a late ascites inflow. The flow of the ascites was convincingly proven by regularly performed colour Doppler UH tests. This old/new method is applicable in patients with therapy-resistant ascites to eliminate the traditional shunt-models. PMID- 12764995 TI - Analysis of serum lipids and lipoproteins in Ethiopian diabetic patients. AB - Serum lipids and lipoproteins were determined in 302 randomly selected diabetic patients attending the Tikur Anbessa Hospital diabetic clinic. The main objective of the study was to analyse lipid levels in type 1 and type 2 diabetic patients. Lipid measurement was done by cholesterol pap method. The mean age was 41.4 +/- 14.4 years (range 14-85 years). One hundred sixty (53%) were males and 142 (47%) were females. There were 140 (46.4%) type 1 and 162 (53.6%) type 2 patients. The mean duration of diabetes mellitus, haemoglobin A1c, fasting blood glucose and random blood glucose were 9.4 +/- 5.4 years, 10.4 +/- 2.2%, 195.5 +/- 79.9 mg/dl and 273.1 +/- 114.5 mg/dl respectively. The mean cholesterol, triglycerides, LDL, VLDL and HDL were 166.5 +/- 45.5 mg/dl, 129.9 +/- 92.4 mg/dl, 94.5 +/- 36.4 mg/dl, 24.4 +/- 15.1 mg/dl and 44.3 +/- 11.5 mg/dl respectively. Hypercholesterolemia and Hypertriglyceridemia were seen in 18.5% and 14.2% of the patients. Total cholesterol was significantly higher in females than in males and in type 2 than in type 1 patients (179.3 +/- 48.4 mg/dl versus 154.1 +/- 38.2 mg/dl, P < 0.01 and 183.2 +/- 43.7 mg/dl versus 145.9 +/- 37.6 mg/dl, P < 0.001) respectively. Triglycerides and LDL cholesterol were also significantly higher in type 2 diabetic patients than in type 1 diabetic patients (162.7 +/- 10.5 mg/dl versus 91.5 +/- 53.3 mg/dl, P < 0.001 and 105.6 +/- 36.2 mg/dl versus 81.9 +/- 32.2 mg/dl, P < 0.001), but HDL cholesterol was the same in both types of diabetic patients. Similarly, hyperlipidemia was associated with obesity and hypertension. The study confirms that lipid values are high particularly in type 2 diabetic patients. Hence our patients are at increased risk of developing atherosclerosis therefore periodic check up of lipids in diabetic patients and effective treatment of the dyslipidemia along with a tight metabolic control was recommended. PMID- 12764996 TI - Delivery of diabetes care in rural Ethiopia: an experience from Gondar. AB - Diabetes is an increasing health problem in African countries. This study was undertaken to ascertain differences in the demography of diabetes in rural and urban populations, and to assess mortality, following a programme aiming to decentralise care to Health Centres nearer to patient's homes. The study population comprised 871 diabetic patients registered in Gondar between June 1995 and August 2000. Four hundred and thirty two were town residents and 439 came from remote rural areas. Most of the rural patients (77%) had Type 1 diabetes whereas in urban areas only 29% had Type 1 and 71% Type 2 diabetes. Review of the mortality of Type 1 diabetic patients up to August 2000 shows that 117 (32%) have died at a mean age of 32.0 +/- 10.4 years, and a duration of diabetes 6.1 +/- 5.0 years. Of the 871 patients, 729 are under regular review from Gondar: the care of 82 percent of the rural patients has been decentralised and is now undertaken in village health centres nearer to their homes. The management of chronic disorders such as diabetes in rural African communities needs to be decentralised to improve access to treatment and reduce mortality. The patients could achieve better health if methods of tracking non-attenders were developed, and if the need to travel large distances at frequent intervals to obtain insulin supplies could be eliminated. PMID- 12764997 TI - Supra-tarsal injection of dexamethasone in the treatment of patients with refractory vernal keratoconjunctivitis. AB - The response to supratarsal injection of dexamethasone in severe refractory vernal keratoconjunctivitis was studied in 20 patients (14 males and 6 females) at the ophthalmology department of Menelik II Hospital, Addis Ababa in 1997. Majority of the patients were in the age range of 11-20 years with mean age of 14.6 years. Although all patients experienced relief of symptoms in the first 2 days of dexamethasone injection, symptoms recurred in the majority 16 (80%) of the patients. Cobble stone papillae and limbal signs disappeared in 2 (11%) and 5 (38%) of the patients respectively. Resolution of limbal form of the disease is more dramatic when compared to the palpebral form. This study showed that supratarsal injection of dexamethasone in severe vernal keratoconjunctivitis is effective in few, partially effective in some and ineffective in many of our patients. Further studies, particularly a randomized controlled trial is recommended prior to utilization of this new therapeutic modality as a standard form of treatment in Ethiopian patients. PMID- 12764998 TI - Beliefs and traditional treatment of malaria in Kishe settlement area, southwest Ethiopia. AB - Kishe settlement area southwest Ethiopia, is endemic for malaria, and malaria related morbidity and mortality are important public health problems. Malaria beliefs and practices are often related to culture, and can influence the effectiveness of control strategies. This study assessed attitude and practices relative to causation, treatment, prevention and control of malaria, and documented traditional malaria treatment practices and remedies, in order to provide baseline data for control program planning and further investigation. A cross-sectional study was conducted in December 1997 by interviewing 254 randomly selected study subjects 85 indigenous and 169 settlers. Eighty three percent of 254 respondents attributed the cause of malaria infection to dirt and rubbish. Ranking of vector control measures was poor, with 77% prioritizing cleaning dirt and rubbish, while only 36% mentioned drainage of swampy areas. Ninety eight percent accepted Dichlorodiphenyltrichlroethane (DDT) house spraying. The prevalence of clinical malaria attacks was 77% and communities had good knowledge about malaria morbidity and mortality. Forty three percent had used traditional medicine for malaria, for reasons including greater accessibility (82%), low cost (48%), lack of awareness about modern medicine (25%) and belief that traditional medicine is better (7%). Most are well informed about malaria morbidity and mortality, understood about the use of DDT spraying and have good treatment seeking behavior but practice of prevention and concept about causation prevention, and control of malaria is poor. It is wise to put emphasis on health education particularly on preventive aspects. Malaria control technicians and environmental health technicians, who closely interact with the community could be used for this purpose. The common traditional treatments for malaria could be further investigated for their effects on malaria parasites and/or symptomatic relief of clinical illness. PMID- 12764999 TI - Perception of dental fluorosis among adolescents living in urban areas of Ethiopia. AB - The objective of the study was to assess the perception of dental fluorosis among adolescents residing in two urban areas in Ethiopia: Addis Ababa (low-fluoride drinking water) and Nazreth (high-fluoride drinking water). Students aged 12-15 years, enrolled in two junior secondary schools in Addis Ababa (n = 161) and two similar schools in Nazreth (n = 177) responded to evaluative statements concerning the appearance of anterior maxillary teeth. Color photographs depicting dental fluorosis (TF score 2, 3, 5 and 7) in maxillary front teeth were used as reference during structured interviews. A majority of the students from Addis Ababa and Nazreth were dissatisfied with the appearance of teeth having TF scores 2 and above. More than 66% of the students from both areas confirmed a need for dental consultation concerning fluorotic teeth with TF > or = [corrected] 2. As compared to their counterparts in Nazreth, children in Addis Ababa felt more embarrassed with TF score 2 and 3 (p < 0.01). TF scores 5 and 7 were found unacceptable both in Addis Ababa and in Nazreth. Students in Addis Ababa, more frequently than their counterparts in Nazreth, believed (erroneously) that dental fluorosis were related to neglect on the part of the child. The present findings confirm that fluorotic teeth constitute a social problem among junior secondary school children residing in low-fluoride as well as in high fluoride urban areas in Ethiopia and, thereby, corroborate the public concern for safe drinking water. PMID- 12765001 TI - A giant retrosternal goiter with severe tracheal compression and superior vena cava syndrome: an operative experience. AB - The peculiarities in the operation of a giant retrosternal goiter with severe tracheal compression and superior vena cava syndrome are highlighted in this report of a 53 year-old female with a large anterior neck swelling interfering with normal breathing and swallowing. From the initiation of the neck incision, mobilization of the gland and performing the subtotal excisions there was troublesome bleeding. Pneumothorax resulting after delivery of the massive retrosternal portion was managed with an underwater-seal drainage tube. PMID- 12765000 TI - The McGill Pain Questionnaire in Amharic: Zwai Health Center patients' reports on the experience of pain. AB - This paper presents for the first time an Amharic translation of the McGill Pain Questionnaire developed by Melzack and used in many countries around the world. It allows for a quantitative and qualitative assessment of the intensity, location, and nature of experienced pain, as well as conditions that relieve pain. Data collected from one hundred patients attending the Zwai Health Center indicated that 81% reported pain at the time, one-quarter of whom were in severe pain. The most commonly chosen descriptors were: burning, stabbing, sore, gnawing, aching, and cramping. Descriptors were often associated with certain diagnoses: burning with gastrointestinal problems, stabbing with respiratory diseases, and gnawing or aching with myalgia/neuralgia. Approximately 40% of those in pain had previously sought relief from a clinic or pharmacy and were attending the center because the pain persisted. Analgesics were more likely to be prescribed for those in mild pain, while other medication without analgesics were prescribed for those in severe pain. The McGill Pain Questionnaire--Amharic (MPQ-Am) could be a useful tool for future studies of illness-specific pain, and of the effectiveness of pharmaceutical and non-pharmaceutical strategies for pain management. PMID- 12765002 TI - Fibrocalcific pancreatic diabetes in a young Ethiopian man. AB - An eighteen year old Ethiopian patient was admitted to Jimma teaching hospital, after presenting with classical hyperglycemic symptoms associated with abdominal pain, chronic diarrhea, cachexia, and radiological appearance of extensive pancreatic calcification. These clinical and radiologic findings correlated with the well-described entity of Fibrocalcific pancreatic diabetes, which has never been reported from Ethiopia before. PMID- 12765003 TI - 26th Kellersberger Memorial Lecture. Lessons from leprosy rehabilitation for general rehabilitation. AB - Leprosy is primarily a disease of skin and peripheral nerves. Because of nerve function impairment, leprosy patients may develop primary nerve related impairments such as, loss of sensation and weakness or paralysis. These primary impairments may lead to secondary impairments such as ulceration and contractures. Many other diseases and disorders present with similar impairments as seen in leprosy e.g. diabetes and peripheral nerve injuries. Nerve function assessment and ulcer prevention and treatment are areas that have been researched in leprosy but these research findings are not yet commonly known and adopted in diseases and disorders that 'relate' to leprosy. Rehabilitation is a relatively new field in medicine and not (well) developed in many developing countries. Rehabilitation requires an integrated approach from different disciplines and professionals. As for other medical specialty fields, rehabilitation demands evidence based practice. PMID- 12765005 TI - Bigger is not always better. PMID- 12765004 TI - Septic shock. AB - Septic shock remains an important cause of death and serious morbidity in medical, surgical and obstetric illness. Many patients with septic shock succumb despite aggressive therapy. Any microorganism can initiate septic shock; the pathophysiology of the disease is not clear; the clinical manifestation is not specific; and there is no reliable laboratory result to diagnose septic shock before it is late. Moreover, despite increasingly complex modes of mechanical ventilation, sophisticated respiratory and hemodynamic monitoring, and a continually expanding array of potent antibiotics, the overall mortality from septic shock remained constant. Better outcome is more likely when recognition is early and treatment is much more aggressive. Therefore, where there is suspicion of septic shock: commence fluid resuscitation, administer oxygen, take blood, urine, and other fluids for culture, commence intravenous broad-spectrum antibiotics, pass a urethral catheter, determine the cause of sepsis and remove if possible, consult for expert medical advice and possible patient transfer to intensive care units, and provide supportive care to involved organ systems. PMID- 12765006 TI - Passive therapeutic gardens. A study on an inpatient geriatric ward. AB - A brief history of the link between horticultural activities and care of patients, particularly psychiatric patients, is reviewed in this article. Past research on both passive and active garden activities is examined in terms of physical and psychological benefits to patients. A passive garden intervention on an inpatient geriatric ward is described. Participants in this study were patients on a geriatric inpatient ward in a mid-sized regional hospital in New Zealand. Behavioral observations of patient movement on the ward were used to demonstrate the effects on patient behavior in response to the presence of the conservatory garden. Results showed a positive reaction to the conservatory, which was maintained 6 months after the initial plants were installed. The benefits of such garden installations are discussed, and areas for further research are outlined. Procedures, ethical concerns, and practical considerations of setting up such a conservatory on an inpatient ward are discussed. PMID- 12765007 TI - Concepts in health promotion. Perceived self-efficacy and barriers in older adults. AB - Many older adults have reached later life in apparent good health while others' lack of self-care activities has greatly accelerated the process of aging. In this article, self-efficacy and perceived barriers are explored as significant concepts that influence the self-care activities of elderly individuals. A conceptual model is presented depicting perceived self-efficacy and barriers as determinants of self-care activities. This conceptual model differs from earlier health promotion models because it is specific for older adults. Based on empirical studies, this conceptual model demonstrates that the variables of perceived self-efficacy and barriers have a profound effect on an older adult's participation in self-care activities. Testing of this model is needed to provide a knowledge base for designing health promotion strategies, maximizing cost containment strategies, and contributing to Healthy People 2010 objectives of increasing quality of life for all older adults. PMID- 12765008 TI - Complementary and alternative medicine. Use in an older population. AB - The aging North American population validates increased research of complementary and alternative medicine (CAM) use by older adults. The purpose of this study was to examine older adults' attitudes and motivations toward CAM use in an attempt to explain its limited usage. Senior citizens (66 to 100 years) were qualitatively surveyed and interviewed to analyze trends in CAM use. Forty-two participants older than 65 completed a questionnaire and 10 of those same individuals participated in an interview session. Motivations for CAM use, prevalence of CAM use, knowledge of CAM, and physician attitudes were investigated. The results of the survey and interviews showed older adults' most prevalent motivations for using CAM were pain relief (54.8%), improved quality of life (45.2%), and maintenance of health and fitness (40.5%). Knowledge of CAM was extremely low across the entire sample, but a significant difference in knowledge level existed among CAM users and nonusers. The CAM therapies most commonly used by older adults were chiropractic (61.9%), herbal medicine (54.8%), massage therapy (35.7%), and acupuncture (33.3%). This sample of senior citizens perceived CAM treatments to be extremely beneficial. Increased education about CAM is needed for older adults and health professionals. Practitioners of CAM should try to understand older adults' motivations for using CAM therapies and be involved in educating older adults about CAM. PMID- 12765009 TI - Walking for exercise self-efficacy appraisal process. Use of a focus group methodology. PMID- 12765010 TI - Common stressors experienced by the well elderly. Clinical implications. AB - The purposes of the article are twofold--first, to report on a study that determined the 10 most commonly experienced stressors in individuals 65 years of age and older, and second, to present nursing interventions for each of the top 10 stressors. The research was conducted on a convenience sample of 200 healthy older adults living in the community using the Stokes/Gordon Stress Scale. The two most commonly experienced stressors were slowing down and concern for world conditions. Others among the top 10 were constant or recurring pain or discomfort, time too short with children or grandchildren, and wishing parts of one's life had been different. Interventions discussed included careful planning of each day's activities, joining discussion groups on world events, participating in a political campaign, using e-mail and other technological communication methods, using reminiscence therapy, and implementing the use of various complementary therapies. PMID- 12765011 TI - A program to reduce fatigue in convalescing elderly adults. AB - This study is an examination of the influence of a fatigue reduction program on the level of fatigue experienced by elderly individuals convalescing in subacute units after lower extremity injury, surgery, or weakness. Fifty participants with a mean age of 81 were divided into two groups of 25 participants. The experimental group received a fatigue reduction program consisting of planned rest periods after bathing and therapy; high-protein, high-carbohydrate liquid supplement; and a 3-minute back rub at bedtime. Each participant's level of fatigue was tested soon after admission and 1 week later, and data were analyzed using the paired t test. The difference between the change in pre-test and post test scores of the two groups was significant in all three measures of fatigue. The experimental group exhibited a significantly greater reduction in fatigue. The outcomes reinforced the value of nursing interventions that have been an integral component of nursing care from its beginning: promoting nutrition, alternating periods of activity and rest, and including a back rub in evening care. PMID- 12765012 TI - Scaled-down nuclease P1 for scaled-up DNA digestion. PMID- 12765013 TI - Real-time assay of tryptase release from human umbilical cord blood-derived mast cells. PMID- 12765014 TI - Double-color fluorescence in situ hybridization with RNA probes. PMID- 12765015 TI - Resistant ribonuclease activity in preparations of total RNA extracted from artiodactyl brain with GITC. PMID- 12765017 TI - PCR product cleanup methods for capillary electrophoresis. PMID- 12765016 TI - Methylation sensitivity of restriction enzymes interacting with GATC sites. PMID- 12765019 TI - Inter-simple sequence repeat-restriction fragment length polymorphisms for DNA fingerprinting. PMID- 12765020 TI - PCR primers containing stop codons reduce the number of false-negatives during blue-white screening. PMID- 12765018 TI - Normalization of luciferase reporter assays under conditions that alter internal controls. PMID- 12765021 TI - Improved method for the construction of full-length enriched cDNA libraries. PMID- 12765022 TI - Enrichment of cells exhibiting tetracycline regulated gene expression. AB - Tetracycline controlled gene expression varies significantly among cells within a cell line. Chromosomal integration sites of the tetracycline transactivator (tTA) gene and/or the test gene presumably account for the variable efficacy of this system. We hypothesized that the efficacy of tetracycline regulated gene expression is more dependent on the level of tTA inside cells and less dependent on the integration sites of the tetracycline transcription units. To test this hypothesis, we established a TetOff regulatied expression of a short-lived enhanced GFP (d2EGFP) via retroviral vectors in a neuroblastoma cell line (NBP2). We then enriched for two populations of NBP2 cells; one expressing high levels of d2EGFP (HG) and the other expressing low levels of d2EGFP (LG) in the absence of doxycycline. We show that the tTA is more abundant in HG cells than in LG cells; the cAMP-mediated transactivation of tTA's promoter further increases the efficacy of the tetracycline system; and the efficient doxycycline regulated expression of a test gene (i.e., VP16CREB) is achieved in HG cells. Therefore, we have developed a simple method to enrich for a population of tetracycline responsive cells with no need for screening for tetracycline-responsive clonal cell lines. PMID- 12765023 TI - High-yield production of recombinant antibody fragments in HEK-293 cells using sodium butyrate. AB - To develop new recombinant monoclonal antibody fragments for therapy and imaging, it is indispensable to have a simple and easy procedure to handle the eukaryotic expression system for production of proteins in high amounts. Gene amplification techniques such as the dehydrofolate reductase (DHFR) system in Chinese hamster ovary cells or the glutamine synthase system in myeloma cells have a couple of disadvantages. The selection procedure is complex, time-consuming, and not fruitful in all cases. The toxic drug methotrexate (for the DHFR system) can increase the production rate but decreases the specific growth rate of the cells. The production rate is not always stable over a long-term cultivation period. To overcome these problems, we are using stably transfected human embryonic kidney (HEK-293) cells in combination with an efficient screening method. Sodium butyrate can increase the expression of recombinant antibody fragments in the transfectomas up to 500 micrograms/4.2 x 10(7) cells/24 h corresponding to 175 micrograms/mL culture medium. This strategy allows a rapid development of new recombinant monoclonal antibody fragments and allows one to proceed rapidly to in vivo testing. PMID- 12765024 TI - Chemical RNA labeling without 3' end bias using fluorescent cis-platin compounds. AB - Recently, fluorescent, monofunctional cis-platin derivatives have been developed to chemically label nucleic acids for use in fluorescent hybridization assays. Here we show by hybridizations to microarrays containing oligonucleotide probes for the 3' ends, middle parts, and 5' ends of mRNAs, that this labeling methodology bypasses the problem of the 3' end bias that is characteristic of the conventional enzymatic oligo(dT)-primed, reverse transcription labeling of mRNAs. PMID- 12765025 TI - Real-time quantitative RT-PCR assay of gene expression in plant roots during fungal pathogenesis. AB - Real-time quantitative RT-PCR is becoming the preferred method for high sensitivity, rapid-throughput RNA transcript quantification. However, due to the significant developmental costs of dedicated fluorogenic probes, a real-time assay that is simple to establish, comparatively inexpensive, and readily adaptable would be advantageous for the detailed analysis of large sets of expressed sequences. We have devised a flexible real-time quantitative RT-PCR assay that employs a nonspecific DNA binding dye for product detection and uses a relative quantification formula to account for differences in PCR amplification efficiency between the target and reference products. The latter permits the use of an exogenous reference transcript and therefore avoids the normal requirement for the construction of a recombinant RNA reference transcript or extensive characterization of housekeeping gene expression. In an investigation of class II chitinase expression in two varieties of Bermuda grass (Cynodon spp.), following infection with the fungal root pathogen Ophiosphaerella narmari, this assay identified 16- and 28-fold peaks in gene expression at 24 and 96 h after inoculation, respectively. PMID- 12765026 TI - Extraction of total RNA from leaves of Eucalyptus and other woody and herbaceous plants using sodium isoascorbate. AB - Rapid extraction of total RNA from Eucalyptus leaves is difficult due to the high content of polyphenolics and polysaccharides. A rapid and simple method was developed by using an extraction buffer containing sodium isoascorbate at a concentration of 500 mM. This method consisted of one or two chloroform extractions, one acid guanidium-phenol-chloroform extraction, and isopropanol precipitation alone. The yields of the RNA fractions were 246-1750 micrograms/g fresh weight when leaves of Eucalyptus, five other woody plants, and four herbaceous plants were used as samples. The contamination of the RNA fractions by proteins and polysaccharides was very limited as judged spectrophotometrically. When the RNA fractions were subjected to agarose gel electrophoresis, intact rRNA bands were detected. The RNA fractions could be used for RT-PCR. These results indicate that our new method achieves a simple and rapid preparation of high quality RNA from leaves of Eucalyptus and other plant species. PMID- 12765027 TI - Kindred DNA amplification from two distinct populations of cDNA fragments. AB - Kindred DNA amplification is a novel and cost-effective method developed to isolate common cDNA fragments between two distinct cDNA populations. Unlike subtractive hybridization, which discards common sequences, kindred DNA amplification isolates and amplifies these sequences within a single hybridization procedure. The utility of this method is demonstrated by cloning the genes in common between two different but metabolically homologous muscles, murine ventricular myocardium and soleus. The reliability of kindred DNA amplification was confirmed by Southern hybridization. PMID- 12765028 TI - Hydrogel-based protein microchips: manufacturing, properties, and applications. AB - Here a simple, reproducible, and versatile method is described for manufacturing protein and ligand chips. The photo-induced copolymerization of acrylamide-based gel monomers with different probes (oligonucleotides, DNA, proteins, and low molecular ligands) modified by the introduction of methacrylic groups takes place in drops on a glass or silicone surface. All probes are uniformly and chemically fixed with a high yield within the whole volume of hydrogel semispherical chip elements that are chemically attached to the surface. Purified enzymes, antibodies, antigens, and other proteins, as well as complex protein mixtures such as cell lysates, were immobilized on a chip. Avidin- and oligohistidine tagged proteins can be immobilized within biotin- and Ni-nitrilotriacetic acid modified gel elements. Most gel-immobilized proteins maintain their biological properties for at least six months. Fluorescence and chemiluminescence microscopy were used as efficient methods for the quantitative analysis of the microchips. Direct on-chip matrix-assisted laser desorption ionization-time of flight mass spectrometry was used for the qualitative identification of interacting molecules and to analyze tryptic peptides after the digestion of proteins in individual gel elements. We also demonstrate other useful properties of protein microchips and their application to proteomics and diagnostics. PMID- 12765029 TI - Specific targeted integration of kanamycin resistance-associated nonselectable DNA in the genome of the yeast Saccharomyces cerevisiae. AB - Sophisticated genome manipulation requires the possibility to modify any intergenic or intragenic DNA sequence at will, without leaving large amounts of undesired vector DNA at the site of alteration. To this end, a series of vectors was developed from a previous gene knockout plasmid system to integrate nonselectable foreign DNA at any desired genomic location in yeast, with a minimum amount of residual plasmid DNA. These vectors have two mutated Flp recognition targets (FRT) sequences flanking the KanMX4 gene and multiple sites for subcloning the DNA fragment to be integrated. The selectable marker can be recycled by Flp site-specific excision between the identical FRTs, thereby allowing the integration of further DNA fragments. With this system, the NLS-tetR GFP and DsRed genes were successfully integrated at the thr1 locus, and the RVB1 gene was tagged at the C-terminus with the V5-epitope-6-histidine tag. This plasmid system provides for a new molecular tool to integrate any DNA fragment at any genome location in [cir+] yeast strains. Moreover, the system can be extrapolated to other eukaryotic cells in which the FLP/FRT system functions efficiently. PMID- 12765030 TI - LacSwitch II regulation of connexin43 cDNA expression enables gap-junction single channel analysis. AB - Metabolic and electrical coupling through gap junction channels is implicated in cell differentiation, tissue homeostasis, and electrotonic propagation of signals in excitable tissues. The characterization of gating properties of these channels requires electrophysiological recordings at both single- and multiple-channel levels. Hence, a system that is able to control connexin expression by external means would provide a useful tool. To regulate the expression of connexins in cells, plasmids encoding a transactivator and/or a lac-operon IPTG response dependent Cx43 target gene were transfected into communication-deficient N2a neuroblastoma cells. Immunoblotting, dye coupling, and electrophysiological methods revealed that expression of Cx43 in selected clones could be tightly regulated. After 15-20 h of acute induction with IPTG, cell-to-cell communication reached its peak with junctional conductances of 15-30 nS. Chronic induction at specific doses of IPTG produced constant, controlled levels of Cx43 expression, which were reflected by predictable junctional coupling levels. These conditions allowed prolonged recordings from either lowly or highly coupled cells, making lac operon an ideal regulatory system for channel gating studies at a single channel level. PMID- 12765031 TI - Antiangiogenic tumor therapy. PMID- 12765032 TI - One-step cellular caspase-3/7 assay. PMID- 12765033 TI - Single nucleotide polymorphism genotyping using allele-specific PCR and fluorescence melting curves. AB - We present a PCR method for identification of single nucleotide polymorphisms (SNPs), using allele-specific primers designed for selective amplification of each allele. Matching the SNP at the 3' end of the forward or reverse primer, and additionally incorporating a 3' mismatch to prevent amplification of the incorrect allele, results in selectivity of the allele-specific primers. DNA melting analysis with fluorescent SYBR Green affords detection of the PCR products. By incorporating a GC-rich sequence into one of the two allele-specific primers to increase the melting temperature, both alleles can be measured simultaneously at their respective melting temperatures. Applying the DNA melting analysis to SNPs in ApoE and ABCA1 yielded results identical to those obtained with other genotyping methods. This provides a cost-effective, high-throughput method for amplification and scoring of SNPs. PMID- 12765034 TI - Lentivirus vector purification using anion exchange HPLC leads to improved gene transfer. AB - Recombinant lentiviral vectors stably transduce both dividing and nondividing cells. Virus pseudotyping with vesicular stomatitis virus envelope G (VSV-G) protein broadens the host range of lentiviral vector and enables vector concentration by ultra-centrifugation. However, as a result of virus vector concentration, contaminating protein debris derived from vector-producing cell culture media is toxic to target cells and reduces the transduction efficiency. Here we report a new and rapid technique for purifying lentivirus vector using the strong anion exchange column that significantly improves gene transfer rates. We purified VSV-G pseudotyped self-inactivating lentivirus vector and obtained two protein elution peaks (Peak 1 and Peak 2) corresponding to transducing activity. Peak 1 viral particles were 4-8 times more effective in transducing target cells than Peak 2 or non-purified (pre-HPLC) viral particles. We used purified lentivirus vector expressing the human Fanconi anemia group A (FANCA) gene to transduce murine hematopoietic stem/progenitor cells. We observed a consistent 2- to 3-fold increase in gene transfer rates using Peak 1 purified virus compared with non-purified virus. We conclude that the purification method using the HPLC system provides the highly purified virus vector that reduces cell toxicity and significantly improves gene transfer in primary cells. PMID- 12765035 TI - Effect of secondary structure on single nucleotide polymorphism detection with a porous microarray matrix; implications for probe selection. AB - Oligonucleotide arrays capable of detecting single nucleotide polymorphisms (SNPs) from amplified nucleic acid have many applications. The expected SNP is usually placed approximately in the center of the probe to ensure the maximum shift in Tm between complementary and SNP sequences. Unfortunately, different short probes (< 30 bases) selected using widely accepted criteria do not perform consistently in this type of assay. Here we present a systematic study on the effect of secondary structure on the ability of oligonucleotide probes to detect an SNP, using real-time array monitoring of a porous microarray substrate that incorporates a novel intra-array mixing system. These results demonstrate that, although positioning of an SNP in the middle of the probe is highly destabilizing, the effect of stable secondary structure on the signal obtained is so dramatic that such probes may be very insensitive. Therefore, if the SNP flanking sequence contains significant secondary structure, then more sensitive probes with good specificity may be obtained by positioning the mutation towards one end of the probe. PMID- 12765036 TI - Using platelet-rich plasma to enhance a composite graft in the maxillary sinus. [Interview]. PMID- 12765037 TI - Child protection. The nurse's role. PMID- 12765038 TI - SARS: current knowledge and management. AB - Severe acute respiratory syndrome (SARS) was identified as a new infectious disease in Asia early in 2003. Since then an unprecedented global public health response has been mounted. This article describes current knowledge of the infection and outlines the management of SARS in the UK. PMID- 12765039 TI - What you need to know about ... asthma. PMID- 12765040 TI - Gastroscopy. PMID- 12765041 TI - Extended independent nurse prescribing in palliative care. AB - Extended nurse prescribing is now a reality and its particular application to specialist palliative care nurses will allow them to formalize their practice of advising doctors on medication regimes and to work more autonomously. An education programme in a validated university is now operating for nurses who wish to become extended formulary nurse prescribers. Extended nurse prescribing for palliative care nurses in the community will be a particular challenge because of the large geographical areas covered, which could make communication with primary care team members difficult. If patients are to receive holistic, seamless care, palliative care nurses need to lobby for an improved, workable formulary. PMID- 12765042 TI - I see diabetes as a lifestyle change and an illness. PMID- 12765043 TI - Nurse-led management of i.v. fluorescein angiography. AB - Fluorescein angiography is undertaken to investigate the state of the circulation in diabetic eye disease, the commonest manifestation of which is diabetic retinopathy. Senior ophthalmic nurses, equipped with the appropriate skills and level of confidence, are now undertaking the procedure. In the eye centre at the Central Middlesex Hospital, a proactive nurse-led approach to the management of patients undergoing fluorescein angiography has had a significant impact on the outcomes of the procedure, and resulted in positive patient feedback. PMID- 12765044 TI - Taking the pain out of waiting. Interview by Leona Armstrong. PMID- 12765045 TI - What valuing people really means. Interview by David Crouch. PMID- 12765046 TI - 'Theatre nursing is in your blood'. Interview by Victoria Hoban. PMID- 12765047 TI - Social psychology of mental health: the social structure and personality perspective. AB - Previous research has revealed a persistent association between social structure and mental health. However, most researchers have focused only on the psychological and psychosocial aspects of that relationship. The present paper indicates the need to include the social and structural bases of distress in our theoretical models. Starting from a general social and psychological model, our research considered the role of several social, environmental, and structural variables (social position, social stressors, and social integration), psychological factors (self-esteem), and psychosocial variables (perceived social support). The theoretical model was tested working with a group of Spanish participants (N = 401) that covered a range of social positions. The results obtained using structural equation modeling support our model, showing the relevant role played by psychosocial, psychological and social, and structural factors. Implications for theory and intervention are discussed. PMID- 12765048 TI - Development of memory structures for homographs using pathfinder network representations. AB - Some studies with children have shown that there is no semantic priming at short stimulus onset asynchrony (SOA) in lexical decision and naming tasks for homographs. The predictions of spreading activation theories might explain this missing effect. There may be differences in children's and adults' memory structures. We have explored this hypothesis. The development of memory structure representations for homographs was measured by a Pathfinder algorithm. In Experiment 1, the three dependent variables were: the number of links in the network, closeness measures (C), and distances between nodes. Results revealed developmental differences in network structure representations in adults and children. In Experiment 2, results revealed that these differences were not due to the cohort effect. In Experiment 3, the relationship between associative strength, as measured by associative norms, and distances, as measured by Pathfinder algorithm, was explored. The results of these three experiments and empirical research from semantic priming experiments show that these differences in memory structure representations could be one of the sources of the missing semantic priming effect in children. PMID- 12765049 TI - Stereotype development in Andalusian children. AB - Social psychologists have shown a profound interest in intergroup relationships, but there are very few papers focusing on the developmental aspects that explain the psychological mechanisms involved in the construction of group and cultural identity. Our research aims to explore how the self-categorization of Andalusian children evolves. We tried to assess the degree to which they self-identify as Andalusian, Spanish, and European, and how this identification changes with age. We were also interested in the affective evaluation of different groups (French, Italian, English, German, Spanish, Catalonian, and Andalusian) made by Andalusian children. Furthermore, we analyzed the relationship between self-categorization and the evaluation of these groups. Results show that the development of national (autonomous community) identity in these children is influenced by their cognitive development, as well as by the relationships among the regional communities of Spain and the relationships between Spain and other countries. The peculiarity of Andalusians as a group is that they assume both identities: Spanish and Andalusian, from a very early age. In-group favoritism is an extended phenomenon at all ages, and Andalusian children have a negative stereotype of the other Spanish groups and other European communities. PMID- 12765051 TI - Alexithymia--state or trait? AB - Alexithymia refers to a specific disturbance in emotional processing that is manifested by difficulties in identifying and verbalizing feelings and a tendency to focus on and amplify the somatic sensations that accompany emotional arousal. Alexithymia is conceptualized both as an affect-deficit disorder and a continuous personality variable. The main purpose of the present study was to investigate the stability levels of alexithymia with regard to changes in emotional distress levels caused by university exams. We tested 20 university students at four different times, before and after the exams. Alexithymic features and self reported emotional distress (trait anxiety and physical symptoms) were measured. Whereas emotional distress measures changed significantly during the diverse phases, the level of alexithymia remained unchanged. We therefore conclude that alexithymia represents a constant trait. PMID- 12765050 TI - Brief-time-series analysis of depressive symptomatology in older people. AB - This research shows the utility of systematic data-gathering from older people and of a statistical analysis procedure for interpreting the data. Four cases of institutionalized older people are presented, and their scores of depressive symptomatology over a period of one to two years is analyzed. Time-series analysis showed a significant positive trend of depression symptomatology in two of the cases, perhaps too subtle for detection in routine clinical check-up, but statistically verifiable. In one of these cases, none of the factors of the scale stands out, despite the fact that the general index shows a statistically significant change over the 36 observations made, so that the results obtained are related to the syndrome of depletion or exhaustion. In the second of the cases, we detect a significant change in the depressive mood factor, which may indicate a subclinical depressive form in its initial stages. Continuous registers can reveal valuable information about situations and progress in the evolution of an older person's mood, with regard to natural development, the prelude to a mood disorder, or follow-up in clinical cases. PMID- 12765052 TI - A new look at cardiac defense: attention or emotion? AB - The study of cardiac defense has a long tradition in psychological research both within the cognitive approach--linked to Pavlov, Sokolov, and Graham's work on sensory reflexes--and within the motivational one--linked to the work of Cannon and subsequent researchers on the concepts of activation and stress. These two approaches have been difficult to reconcile in the past. We summarize a series of studies on cardiac defense from a different perspective, which allows integration of the traditional approaches. This new perspective emphasizes a sequential process interpretation of the cardiac defense response. Results of descriptive and parametric studies, as well as those of studies examining the physiological and psychological mechanisms underlying the response, show a complex response pattern with both accelerative and decelerative components, with both sympathetic and parasympathetic influences, and with both attentional and emotional significance. The implications of this new look at cardiac defense are discussed in relation to defensive reactions in natural settings, the brain mechanisms controlling such reactions, and their effects on health and illness. PMID- 12765053 TI - [Prostate cancer screening]. AB - Prostate cancer has become the most frequent cancer and the second cause of cancer mortality in men. This public health problem is becoming increasingly important due to the increasing life expectancy. At the present time, prostate cancer will be discovered in one in every eight men during their lifetime. Prostate cancer represents 25% of all new cases of male cancers. Prostate cancer screening is designed to detect early stage, asymptomatic prostate cancer, as the patient's chances of cure are higher when the cancer is diagnosed at an early stage. The conclusions of the ANAES evaluation in 1998 did not recommend mass screening for prostate cancer. Several international prospective randomized studies based on serum PSA assay, sometimes associated with digital rectal examination, are currently underway. France is participating in the European ERSPC study (European Randomized Study of Screening for Cancer Prostate) and is organizing a national study on high-risk populations. While waiting for the final results of these studies, a recommendation needs to be proposed to inform general practitioners and specialists about optimal use of the currently available tests. Based on the conclusions of its oncology committee (composed of urologists, medical oncologists, radiotherapists, pathologists and radiologists), the Association Francaise d'Urologie proposes a recommendation concerning prostate cancer screening and defines its modalities, especially concerning the target population, screening tests and the information given to men before screening. The Association Francaise d'Urologie recommends prostate cancer screening by PSA assay (prostate specific antigen) and digital rectal examination annually between the ages of 50 and 75 years, and from the age of 45 years in men with a family or ethnic risk. If total PSA is above the normal value of the test or if digital rectal examination is abnormal, referral to a urologist is recommended. Information concerning the limits, benefits and risks of screening and the available treatment options must be given before performing these examinations. PMID- 12765054 TI - [Retroperitoneal laparoscopic nephrectomy fo polycystic kidney: preliminary results]. AB - OBJECTIVES: To evaluate the morbidity and mortality of retroperitoneal laparoscopic nephrectomy for polycystic kidney. MATERIAL AND METHODS: Between June 2000 and March 2002, seven retroperitoneal laparoscopic nephrectomies for polycystic kidney were performed in six patients (three men, three women) with a mean age of 52.8 years. All patients presented end-stage renal failure treated by haemodialysis in five cases and by renal transplantation in one case. The ASA score was 2 in four cases and 3 in two cases. The indication for surgery was preparation for renal transplantation, episodes of macroscopic haematuria and pain in three cases, hypertension poorly controlled by medical treatment in two cases, preparation for renal transplantation in one case and pain associated with restrictive respiratory syndrome due to compression in one case. RESULTS: The mean operating time was 4 hours 35 minutes, and the mean blood loss was 400 ml. There were no intraoperative or perioperative deaths. An early postoperative complication occurred in two cases. The mean hospital stay was 11 days, with a mean stay of 2.4 days in the postoperative intensive care unit. No late complications were observed with a mean follow-up of 14 months. Preoperative pain, episodes of haematuria, hypertension and signs of compression resolved in each case. CONCLUSION: Laparoscopic nephrectomy for polycystic kidney is a technique that can be performed via a retroperitoneal approach without manual assistance and with low morbidity. PMID- 12765055 TI - [Value of the uCyt+ test for the detection and followup of bladder tumors]. AB - INTRODUCTION: Superficial bladder tumours are common tumours that frequently relapse, generally with deterioration of the grade. Cystoscopic surveillance is unpleasant for the patient and is associated with considerable morbidity, while urine cytology, although very easy to poor, has a low sensitivity. According to the literature, the uCyt+ test is very effective for the detection of urothelial tumours. This immunofluorescent technique detects desquamated tumour cells previously labelled by specific antibodies. MATERIAL AND METHOD: This prospective study was conducted in 92 patients undergoing screening or follow-up cystoscopy. Voided urine was collected prior to cystoscopy to perform standard cytology and a uCyt+ test. These examinations were interpreted independently under blinded conditions. RESULTS: The sensitivity and specificity of uCyt+ were 68.7% and 81.7%, versus 40.6% and 96.7% for cytology. The sensitivity of uCyt+ was higher for low-grade tumours (87.5%), in contrast with cytology (37.5%). The results of uCyt+ were comparable to those of cytology for high-grade tumours (80%). CONCLUSION: The uCyt+ test presented better performances than standard cytology. It would allow less frequent surveillance cystoscopy examinations, especially in patients with low-grade lesions. PMID- 12765056 TI - [Bladder tumor in young patients: a series of 26 cases. Comparison with a review of the literature]. AB - OBJECTIVE: Urothelial bladder tumours are rare before the age of 40 and represent only 1% to 4% of all bladder tumours. This study reviews the histological features of these tumours and tries to more clearly define their natural history. MATERIAL AND METHOD: From 1980 to 2000, we observed 2,600 cases of primary bladder tumours; 34 subjects were younger than 40. We studied the course of 26 subjects with a urothelial tumour. Their mean age was 30 years (range: 16 to 40 years) and 8 patients were younger than 30. RESULTS: 22 of the 26 patients presented a superficial tumour, including 21 cases of stage Ta tumour. The majority of tumours were well differentiated. Two patients had a multifocal tumour and 4 patients had a tumour larger than 3 cm in diameter. The overall recurrence rate was 55% with progression of the tumour stage in 9% of cases. A better prognosis was observed before the age of 30, with a recurrence rate of 38% versus 62% in patients between the age of 30 and 40. Four patients had an invasive tumour, which was metastatic at the time of diagnosis in 3 cases. CONCLUSION: The natural history of superficial bladder tumours appears to differ according to age. Before the age of 30 years, tumours are frequently stage Ta, well differentiated, with a low recurrence rate. Between the ages of 30 and 40 years, the course of superficial bladder tumours is similar to that observed in older subjects. However, the prognosis of invasive tumours is usually very poor, indicating a particularly aggressive potential. PMID- 12765057 TI - [Value and justification of urine dip-sticks in the diagnosis of postoperative urinary infections in urology]. AB - OBJECTIVE: To assess the diagnostic value of urine dip-sticks for postoperative urinary tract infections. MATERIAL AND METHODS: A prospective study was performed on 165 patients undergoing radical prostatectomy, open prostatectomy for BPH or transurethral resection of the prostate or bladder. Postoperative bacteriuria was investigated by using Multistix urine dip-sticks and was verified by urine culture. The presence of nitrites was the only parameter of the dip-stick taken into account. RESULTS: The overall infection rate was 9.7% (range: 7.3% to 16%). The dip-stick showed extremely poor sensitivity and specificity of 36.3% and 57.8%, respectively (p = 0.03). The main bacterial species isolated were: Escherichia coli (63%) and Enterococcus faecalis (27%). CONCLUSION: The lack of diagnostic reliability of urine dip-sticks no longer justifies the use of these tests for the diagnosis of postoperative infection. Urine culture, with documented efficacy, remains the reference examination. PMID- 12765058 TI - [Detection of telomerase activity by semi-quantitative and in situ assays and quantification of hTERT expression in bladder carcinomas]. AB - OBJECTIVES: Several studies have reported the important role played by telomerase, an enzyme which maintains the length of telomeres, during carcinogenesis. The objective of this study was to detect telomerase activity by semiquantitative in situ methods and to quantify expression of the hTERT subunit in a group of bladder cancers in order to assess its diagnostic and prognostic value. MATERIAL AND METHODS: Telomerase activity (TA) was detected by the TRAP method ("telomeric repeat amplification protocol") on a series of 29 bladder cancers and 3 samples of healthy mucosa. Levels of expression of the hTERT gene were studied by real-time quantitative RT-PCR. In situ detection of TA was performed by using the same kit as the TRAP method, applied to frozen tissue sections exclusively for cases with discordant results between TA detection and hTERT mRNA. RESULTS: TA was detected by TRAP in 15 of the 29 bladder cancers. hTERT mRNA was detected and quantified in all tumour samples. A significant correlation was observed between TA and the level of hTERT mRNA expression. In situ detection of TA demonstrated heterogeneous TA in tumour tissue. A significant correlation was observed between the level of hTERT mRNA expression and tumour histological grades and stages. CONCLUSION: hTERT mRNA detection by real-time quantitative RT-PCR appears to be a sensitive method for the diagnosis of bladder cancer with a good correlation with tumour grade and stage. Quantitative evaluation of hTERT mRNA could therefore be a useful marker of tumour progression for the early diagnosis and follow-up of bladder cancers. PMID- 12765059 TI - [Association between neurologic involvement and lower urinary tract dysfunction and their symptoms in patients with multiple sclerosis]. AB - OBJECTIVE: To establish an association between different types and degrees of neurological involvement and lower urinary tract dysfunctions and their symptoms in patients with multiple sclerosis. PATIENTS AND METHODS: The studied group comprises 84 patients with multiple sclerosis with an average of 42.1 years in whom neurological and urological functional involvement had been evaluated. Neurological involvement had been evaluated using Expanded Disability Status Scale (EDSS) score; urinary tract dysfunctions have been diagnosed by urodynamic examinations and lower urinary tract symptoms (LUTS) using the micturition questionnaire. RESULTS: 1) An association between neurological involvement and lower urinary tract dysfunctions: detrusor hyperreflexia depends on the degree of pyramidal system involvement and the presence of detrusor-sphincter dyssynergia depends on overall disability and the degree of pyramidal system involvement. 2) An association between lower urinary tract dysfunctions and LUTS: obstructive symptoms depend on the presence of detrusor-sphincter dyssynergia. 3) An association between neurological involvement and LUTS has not been established. CONCLUSION: An association between neurological involvement, lower urinary tract dysfunctions and their symptoms has been found. Based on this knowledge, it is possible to simplify the diagnostic and therapeutic approaches in certain groups of patients. PMID- 12765060 TI - [PSA: the difficult position of general practitioners between patients and urologists]. AB - OBJECTIVE: To determine the reasons for ordering PSA assay and delayed referral to an urologist following detection of elevated PSA in general practice. MATERIAL AND METHOD: Retrospective study of 200 patients referred to the department by general practitioners for elevated PSA from September 2000 to April 2001. RESULTS: The median age was 67 years (range: 52 to 87). The median PSA was 7.9 ng/ml (range: 4.1 to 897). Patients were referred after 1 to 5 PSA assays (mean: 1.6) with a median interval after discovery of elevated PSA of 1.7 months. The referral time was greater than 6 months for 52 patients (26%) and 31 patients (15.6%) were referred after more than two PSA assays were found to be elevated. PSA assay was ordered in the absence of symptoms or abnormal digital rectal examination in 43% of patients over the age of 70 years. CONCLUSION: Prostate cancer screening is regularly performed in general practice. The information given to general practitioners, faced with a well informed population, appears to be insufficiently clear, as PSA assay is sometimes incorrectly ordered or interpreted. PMID- 12765061 TI - [UCLA/RAND Cancer Prostate Index quality of life questionnaire after external beam radiotherapy for localized prostate cancer: repercussions of complications and quality of life in general]. AB - INTRODUCTION: This study was designed to evaluate the quality of life after external beam radiotherapy for localized prostate cancer using the UCLA/RAND Cancer Prostate Index questionnaire. MATERIAL AND METHODS: An accurate translation of the questionnaire was retrospectively sent to 108 patients treated for localized prostate cancer by exclusive external beam radiotherapy between 1989 and 1999. The reference values adopted for comparison were those observed by Litwin in a control population without prostate cancer. Patient subgroups were constituted according to the presence or absence of neoadjuvant endocrine therapy and laboratory signs of progression (ASTRO). RESULTS: The response rate was 61.1%, the mean age of the patients was 71.9 years and the mean follow-up was 46.5 months. The percentage of patients "living in a couple" or with "a serious relationship" was 93.4% Three quarters of the population declared that they were "satisfied" or "very satisfied" with the treatment performed. General quality of life scores were comparable to those of Litwin's control population. A functional alteration and decreased tolerance of impairment were observed, in decreasing order, for the sexual, gastrointestinal and urinary factors. CONCLUSION: External beam radiotherapy essentially alters quality of life related to gastrointestinal and sexual functions, while the only aspect of urinary function studied by this questionnaire is continence. PMID- 12765062 TI - [Complications of circumcision. Report of 63 cases]. AB - OBJECTIVE: The aim of this study was to report complications of circumcision received in urologic center. MATERIAL AND METHODS: From a retrospective study during a period of 11 years, 63 cases of complications were noticed secondary to traditional circumcision (n = 45) or paramedics (n = 18). RESULTS: The different complications were: urethral fistulas (n = 41), total or partial glans amputation (n = 9), urethral stricture of the meatus (n = 8), penile pseudo elephantiasis (n = 2), penile denudation (n = 1), hemorragiae (n = 6) and septicaemia (n = 1). The authors underline clinical aspects of these complications and the problems of their treatment. PMID- 12765063 TI - [Nephroblastoma and Wilms tumor: report of 2 cases]. AB - Nephroblastomatosis is a rare disease, considered to be a precursor of Wilms tumour. When it is isolated, recommended treatment consists of chemotherapy followed by surveillance. In the presence of associated Wilms tumour, treatment consists of preoperative chemotherapy followed by partial nephrectomy (when possible), followed by postoperative chemotherapy and surveillance. PMID- 12765064 TI - [Louis Ombredanne (1871-1956) and the cure for hypospadias]. AB - Ombredanne was one of the founders of paediatric surgery. He was the inventor of surgery technics. His methods used in infantile urology became classics, such as hypospadias' operation. PMID- 12765065 TI - [Xanthrogranulomatous pyelonephritis with nephrocutaneous fistula]. AB - The authors report a case of xanthogranulomatous pyelonephritis presenting with nephrocutaneous fistula. This case illustrates all of the typical features of this disease: late diagnosis, non-functioning affected kidney, staghorn calculi, urinary tract anomaly, perinephritis with fistulization. The authors review the diagnostic and therapeutic modalities of xanthogranulomatous pyelonephritis and discuss the other aetiologies of nephrocutaneous fistula. In view of the severity of this disease and its preoperative resemblance to renal cancer, nephrectomy is often the only available treatment option. PMID- 12765067 TI - [Leiomyosarcoma of the inferior vena cava: diagnostic problem and treatment]. AB - The authors report the case of a 42-year-old woman in whom a tumour of the inferior vena cava was discovered intraoperatively. Histological examination of the specimen confirmed the diagnosis of leiomyosarcoma. Malignant tumours of the inferior vena cava are very rare and about 95% of these tumours are leiomyosarcomas. These tumours are diagnosed preoperatively in only 10% of cases. CT and MRI allow a precise staging assessment. Radical surgery and a combination of radiotherapy-chemotherapy constitute optimal treatment for leiomyosarcoma of the inferior vena cava, ensuring long survival, but recurrences are frequent. PMID- 12765066 TI - [Stauffer syndrome associated with a giant renal tumor]. AB - The authors report a new case of Stauffer syndrome characterized by cholestatic jaundice in a 54-year-old patient with renal tumour. This paraneoplastic syndrome resolved after nephrectomy. The authors discuss the pathophysiological, diagnostic and therapeutic aspects in the light of a review of the literature. PMID- 12765068 TI - [Bladder amyloidosis. Report of two cases]. AB - Bladder amyloidosis is a rare disease. The clinical features cannot be distinguished from those of urothelial tumours. The authors report two cases of type AL bladder amyloidosis associated with multiple myeloma and discuss the aetiology, histology and clinical course of this disease. PMID- 12765069 TI - [Malignant transperitoneal bladder rupture]. AB - The authors report a case of malignant intraperitoneal bladder rupture due to a stage PT1 grade II bladder tumour. The clinical features, mechanism of rupture, treatment and subsequent follow-up are discussed in the light of a review of the literature. PMID- 12765070 TI - [Intraperitoneal bladder rupture during transurethral resection of the prostate]. AB - Bladder explosions are a rare complication of endoscopic surgery caused by massive cellular hydrolysis during electrocoagulation, releasing a mixture of gases that becomes explosive in contact with oxygen. Oxygen may enter the bladder as a result of manipulation of the resector, incorrect use of the Ellick evacuator bulb or introduction of air bubbles via the irrigation tubing. This risk can therefore be limited by taking certain precautions. The authors report a new case of intraperitoneal bladder rupture during transurethral resection of the prostate. PMID- 12765071 TI - [Pre-pubic abscess of unknown cause]. AB - The authors report a case of prepubic abscess in a young man presenting with pain. Surgical exploration revealed a group A Streptococcus abscess. The cause of this abscess was not determined. PMID- 12765072 TI - [Prostate cancer and acute disseminated intravascular coagulation. Therapeutic management based on three cases]. AB - Acute DIC is a rare, but life-threatening complication of metastatic prostate cancer. The authors discuss the treatment modalities in the light of three cases and a review of the recent literature. The key to treatment of DIC is treatment anti of the tumour. Androgen blockade is indicated in hormone-dependent tumours. This treatment can sometimes be completed by low-dose oral anticoagulants. Chemotherapy is the treatment of choice of acute DIC during the hormone resistance phase. PMID- 12765073 TI - [Prostatic cancer in the young adult]. AB - Prostatic carcinoma is exceptional in young adults before the age of 30. It is often diagnosed at a late, clinically advanced stage with a poorly differentiated histological type. Treatment is usually palliative and the prognosis is very poor with a mean survival of 6 months. The authors report a new case in a 25-year-old patient with locally advanced poorly differentiated prostatic carcinoma and a Gleason score of 10 treated by endocrine therapy, radiotherapy and chemotherapy with a good clinical and radiological course with a follow-up of two years. PMID- 12765074 TI - [Invasive angiomyxoma]. AB - Invasive angiomyxoma is a rare mesenchymal tumour predominantly affecting women and essentially involving the pelvis, perineum and inguinal region. It is essentially characterized by a high local recurrence rate. Treatment of these recurrences is almost exclusively surgical and is not always easy to perform. Other treatment modalities have been proposed (radiotherapy, endocrine therapy) with encouraging preliminary results. The authors report the case of a patient presenting with a recurrence at 9 months, treated by surgery and preoperative embolization. No disease progression was observed at the various follow-up CT examinations 9 months after the operation. Along with most other authors, they believe that surgery is still the standard treatment for these recurrences, but this attitude may need to be revised with the appearance of new treatment modalities. PMID- 12765075 TI - [Rare differential diagnosis of urinary incontinence]. AB - The authors report the case of a 55-year-old woman with prolapse presenting a differential diagnosis of urinary incontinence: a peritoneo-vaginal fistula with serous discharge in a patient with ascites and a history of hysterectomy. The only cases of peritoneo-vaginal fistula reported in the literature were discovered during extra-uterine pregnancy after hysterectomy. PMID- 12765076 TI - [Testicular infarction without torsion in cryptorchism]. AB - Testicular infarction without torsion of the spermatic cord is a rare lesion, generally idiopathic and spontaneous. It may be exceptionally associated with certain risk factors (vasculitis, cholesterol emboli, malakoplakia, protein S or antithrombin III deficiency, etc.). It often simulates testicular tumour, leading to orchidectomy, which can be avoided, particularly in the case of segmental infarction, by applying conservative treatment. The authors report an original case of infarction of an inguinal ectopic testis without torsion. No cause was identified. In the light of this case and a review of the literature, the authors discuss the various aspects of this clinical entity. PMID- 12765077 TI - [Schwannoma of the penis: a rare and unusual tumor of the external genital organs]. AB - The authors report an isolated case of schwannoma of the penis in a 34-year-old man. The diagnosis was based on precise histological and immunohistochemical criteria and treatment consisted of simple surgical excision followed by clinical follow-up. PMID- 12765078 TI - [Urologic presentation of acute appendicitis: a frequent cause of delayed diagnosis, including in HIV infected patients]. AB - Acute appendicitis rarely presents in the form or urinary tract symptoms. Atypical forms of acute appendicitis occur more frequently in subjects infected by the human immunodeficiency virus (HIV). The authors report a case of acute appendicitis in an HIV-infected subject mimicking acute pyelonephritis secondary to obstruction. Atypical forms of acute appendicitis must be diagnosed and treated as early as possible. Computed tomography plays an essential role in the diagnostic assessment of abdominal and lumbar symptoms in HIV-infected patients. PMID- 12765079 TI - [Urological manifestations of Behcet's disease]. AB - Behcet's disease is a systemic disease most frequently presenting in the form of oro-genital ulcers. More rarely, other genital and bladder lesions, responsible for disorders of micturition, may also be observed. In these cases, urological management is necessary to prevent repercussions on the upper urinary tract. The authors report the case of a young patient who presented with epididymitis and severe urinary frequency in the context of this disease. The various clinical signs responded to simple medical treatment. PMID- 12765080 TI - [Scrotal calcinosis]. AB - The authors report a case of scrotal calcinosis in a 56-year-old patient. In the light of a review of the literature, they discuss the aetiopathogenic, clinical and histological features of this rare disease. PMID- 12765081 TI - [Hormonal therapy in metastatic prostatic cancer]. AB - For more than sixty years, the treatment of advanced prostate cancer has been based on androgen deprivation. Despite this long follow-up, the optimal treatment regimen is still a subject of controversy. Treatments inducing chemical castration have been greatly improved over recent years with the appearance of LH RH analogues, non-steroidal antiandrogens and finally, more recently, LH-RH antagonists and Gn-RH agonists. The modalities of endocrine therapy have also evolved: early or late, total, intermittent or even targeted to the prostate by the use of non-steroidal antiandrogens alone. In this article, the authors review the results of each of these various treatment options in order to place them in perspective with the real benefit that can be expected in terms of quality of life and survival in patients with metastatic disease. PMID- 12765082 TI - [One-stage laparoscopic bilateral upper pole nephrectomy for ectopic insertion of complete ureteral duplication]. AB - Ectopic ureteric insertion of the superior renal unit in the context of complete ureteric duplication is frequently accompanied by destruction of the corresponding parenchymal territory. Surgical treatment must comprise upper pole nephrectomy with partial ureterectomy. The authors report a case of one-stage retroperitoneal laparoscopic bilateral treatment which could become the treatment of choice of this disease. PMID- 12765083 TI - [Re "Penile prosthesis and infections" by Yann Schoepen, Frederic Staerman. Prog. Urol., 2002, 12, 377-383]. PMID- 12765084 TI - Short-term memory, morphology, and reading. PMID- 12765085 TI - Job satisfaction: perceptions of a national sample of teachers of students who are deaf or hard of hearing. AB - The study examined the perceptions of a national sample of teachers of students who are deaf or hard or hearing to assess their level of job satisfaction. A questionnaire was developed and distributed; 610 completed surveys were analyzed. Overall, respondents appeared satisfied with their jobs. Of the 59 items in the survey, 51 were scored as positive for the group as a whole. Participants reported that their relationships with colleagues were the most enjoyable aspect of the job. Paperwork, state assessment tests, and lack of family involvement were identified as the least satisfying aspects. Data were also analyzed by comparing the responses of teachers across groups--itinerant, elementary, secondary, and resource room. Generally, this group-by-group analysis produced findings similar to those for the overall sample. Recommendations on addressing the specific factors that teachers responded to negatively are provided. PMID- 12765086 TI - Deafness in sub-Saharan Africa. AB - Deaf education in sub-Saharan Africa originated in the 19th century, primarily through efforts by hearing European missionaries who typically followed their homelands' oral-only practices. But education became available to only a fraction of the deaf population. In the 20th century, Andrew Foster, a deaf African American missionary and Gallaudet University's first African American graduate, had unparalleled impact on deaf education in the region, establishing 31 schools for the Deaf, training a generation of deaf leaders, and introducing his concept of Total Communication, which embraced both American and indigenous signs. Nigeria, Kenya, Uganda, and South Africa have provided leadership in deaf education, but throughout the region there is growing acceptance of sign language use in school, and secondary and postsecondary education for the Deaf is increasingly available. Some national constitutions safeguard the rights of citizens with disabilities and even recognize indigenous sign languages. International disability organizations, particularly the World Federation of the Deaf, have helped change attitudes and train leaders. Despite some grim present realities, prospects for continued progress are good. PMID- 12765087 TI - Preparing today's teachers of the deaf and hard of hearing to work with tomorrow's students: a statewide needs assessment. AB - The Individuals with Disabilities Education Act is explicit in its mandate that students who receive special education services have opportunities to be involved in and progress in the general education curriculum. Teachers providing instruction to students who are deaf or hard of hearing are expected to comply with this federal mandate. To determine if teachers of students who are deaf or hard of hearing throughout the state of Georgia felt adequately prepared to educate this population, a statewide needs assessment survey was conducted. Questionnaires were reviewed from 110 experienced teachers of students who are deaf or hard of hearing. More that half of the teachers who responded judged their teacher preparation program to be appropriate. Specific suggestions for modifications to teacher preparation programs are provided. PMID- 12765088 TI - Deaf college students' perspectives on literacy portfolios. AB - The study examined how literacy portfolios were used as tools in a college developmental English class in which deaf students assessed their reading comprehension as well as their writing processes and products. The students' reading and writing assignments involved reflective thinking and were grounded in authentic tasks. Immediate feedback was provided. The study was multidimensional, longitudinal, and ongoing. A variety of field research techniques were used to ascertain the uses and influences of portfolios in regard to students' reading, writing, and reflective thinking. The results support the idea that the use of literacy portfolios can positively influence students who are deaf when they assess their reading and writing abilities. PMID- 12765089 TI - School-to-work experiences: curriculum as a bridge. AB - Because workers must be prepared to constantly update their skills and reeducate themselves for new skills, lifelong learning must be a goal for every teacher and student. The curriculum provided during educational experiences should pave the way for attainment of this goal. This is particularly true for deaf and hard of hearing students. School-to-work (STW) has become an umbrella term for activities, experiences and opportunities that prepare students for the world of work, such as youth apprenticeships, mentoring, internships, job shadowing, career exploration, and integration of academic and vocational curriculum. This article provides historical background on STW, laws shaping requirements for STW programs, and research supporting STW components. Recommendations are provided for curricular elements of an STW program, and trends in STW are forecast. PMID- 12765090 TI - Superintendents of American residential schools for the deaf: a profile. AB - Data on selected characteristics of superintendents of American residential schools for the Deaf were gathered in a 1999 survey. The resulting profile of superintendents of residential schools was then compared with a profile of superintendents of public elementary and secondary schools that had been compiled in 1992 by the American Association of School Administrators. The study population consisted of the lead administrators of the 72 residential schools for the Deaf in operation in the United States at the time of the survey. One particularly note-worthy finding was that these superintendents reported the same beliefs about their essential responsibilities that had been reported by superintendents of public elementary and secondary schools. The greatest disparity between the two groups of superintendents was in how they characterized their relationships with their governing boards: Generally, the residential school superintendents reported relationships that were less formal. The study, apparently the first attempt to profile superintendents of American residential schools for the Deaf, establishes a baseline for future studies of this kind. PMID- 12765091 TI - Deaf and hard of hearing students' performance on arithmetic word problems. AB - There has been limited research into the intersection of language and arithmetic performance of students who are deaf or hard of hearing, although previous research has shown that many of these students are delayed in both language acquisition and arithmetic performance. The researchers examined the performance on arithmetic word problems of deaf and hard of hearing students in the South East Queensland region of Australia; they also examined these students' problem solving strategies. It was found that performance on word problems was similar for deaf and hearing students, but that deaf students experienced delays in achieving successful performance on word problems relative to their hearing peers. The results confirm the findings of other studies showing that students who are deaf or hard of hearing experience delayed language acquisition, which affects their capacity to solve arithmetic word problems. The study conclusions stress the need for greater use of direct teaching of analytic and strategic approaches to arithmetic word problems. PMID- 12765092 TI - A review: neural control of mastication in humans as influenced by food texture. AB - This review summerizes recent approaches to the physiology of the masticatory system in humans that aim to understand how the process is influenced by the material properties of foods. The centerpiece is a group of experiments that show that the rate of breakdown of food in human mastification depends principally on the combination of two mechanical properties of foods: toughness(R) and modulus of elasticity (E). Two mechanical indices are constructed from these properties: the square root of their product, (ER)0.5, is predicted to explain the resistance to an incisal bite, while the square root of their ratio, (R/E)0.5 is predicted to control the rate of fragmentation during a postcanie bite. Evidence for the latter is reviewed, which also appears to modulate the activity of jaw closing muscles and the extent of lateral mandibular movement during mastication. These studies provide a quantified link between the food stimulus and the physiological response of the mastiatory system for which we know of no parallel. Attempts to extend this analysis have been made by psychophysical investigations of food texture. These support some sensitivity to the mechanical index that we have identified, but are not conclusive. Finally, we provide a chart summarizing physiological responses to food texture that could interest dentists, food scientists and also those interested in the analysis of dentition and diet in mammals. PMID- 12765093 TI - Position of mandibular condyle at maximal mouth opening in symptom free subjects. AB - The purpose of this study was to ascertain maximal mouth opening in normal symptom free subjects and to evaluate radiographically the anatomic position of the condyle during such an opening. Also to find out the prevalence of clinical signs of TMJ dysfunction in asymptomatic subjects. Sixty subjects aged 15-45 years were selected according to stringent criteria. The clinical evaluation comprised of measurements of mouth opening, deviation on opening and assessing any joint sounds. Then transcranial radiographs were taken and different linear measurements were done. The values and correlations were analyzed using students 't' test. Females presented higher degree of clinical signs (joint sounds) of TMD than males. Almost all the subjects had their condyles translated beyond the articular eminence showing more of an inferior vertical shift. It is required that similar studies be conducted on different ethic population and different age groups. Results could be interesting and shed light in the relativity dark area of TMD. PMID- 12765094 TI - Mandibular canine index in establishing sex identity. AB - An investigation study on sex identity through mandibular canine index directed to detect sexual dimorphism using the Mesio-Distal width of mandibular permanent canines and inter canine and inter canine arch width in the mandible was conducted in the Department of Oral Medicine and Radiology, Bapuji Dental College and Hospital, Davangere. 360 patients were subjected to the mesio-distal measurement and inter canine arch width. Males were detected correctly in 83.3% and in females 81%. They were statistically significant and the related literatures reviewed. PMID- 12765095 TI - Shear bond strength of 4th & 5th generation dentin bonding agents in the presence and absence of moisture. An in vitro study. AB - Extracated human molars were used for the study, flat dentinal surfaces were prepared on the occlusal surfaces, and then 4th and 5th generation dentin bonding agents were used in dry and moist conditions to bond composite resin onto the flat dentin surfaces. Shear Bond Strength was measured using Instron Universal Testing machine. Failures were recorded as adhesive, cohesive and mixed. Bond strength was highest in moist condition, because the moist surface permits more porous collagen network and hence greater infiltration of adhesive monomers, than do surfaces that are air dried. Moreover, those dentin bonding agents whose primers are based on acetone show improved bond strength, because acetone aggressively pursues and displaces water in the substrate, resulting in the primer resin being carried into tissue channels and porosities. When the dentin is air dried, the water that is supporting the collagen network evaporates causing the collagen network to collapse network is greatly reduced, which in turn decreases the premeability of intertubular dentin to adhesive resin and as a result causes decrease in bond strength. 5th generation dentin bonding agent showed significant improvement in the moist dentin bond strength. PMID- 12765096 TI - Oral health status and treatment requirments of confectionery workers in Bangalore city. A comparative study. AB - The present study was undertaken to assess the oral health status and treatment requirement of cofectionery workers and also to assess the influence of confectionery environment ion their oral health. A total of 502 confectioners were examined and studied under 4 groups, Biscuits gp[107], chocolate gp[160], sweets gp[144] and Bakery gp[91]. Age and sex matched 294 wheat flour workers served as controls. 60.36% of confectioners exhibited caries experience with significantly higher DMFT compared to controls. Among confectioners sweets and chocolate groups had higher caries experience. Increase in caries experience with increase in duration of employment among confectioners were observed. Also confectioners consuming confectionery products more than 6 times/day had significantly higher caries experience than those consuming 0 to 3 and 4 to 6 times/day. Periodontal diseases was significantly higher among confectioners than controls more so in sweets and biscuit groups. Further extensive unmet treatment needs like oral prophylaxis, periodontal therapy, extractions, prosthetic rehabilitations were observed among confectioners. PMID- 12765097 TI - Oral health status among the tobacco and betel nut chewers in the Kishore Ganj District of Bangladesh. A statistical study. AB - A survey was taken among the people of the Kishore Ganj District in Bangladesh regarding their mouth status. A deep analysis was made with respect to the most common habit prevalent among the people namely "Tobacco and Betel nut chewing". This habit is prevalent among both the men and women in that district. The oral manifestations of 780 people were studied in depth and the results noted. PMID- 12765099 TI - A review of odontogenic keratocyst with a report of unusual occurrence in the maxilla. AB - Odontogenic keratocysts (OKC) are keratinizing epithelium-lined cycts of the jaws with well-defined histologic criteria and possessing one clinical feature warranting their recognition and separation as a distinctive entity, due to their exceedingly high recurrence rate. This report describes a case of OKC which occurred in the anterior maxilla in a 12 year old female child, with its treatment. These lesions are normally seen in the mandible, and are rare in the maxilla. PMID- 12765098 TI - A clinical and SEM evaluation of the efficiency of sofscale gel and hand scaling and hand scaling alone. AB - The purpose of this study is to compose between hand scaling with abd without the calculus solvent gel (sofscale) and ultrasonic instrumentation at clinical and SEM level. 30 patients belonging to the age group of 17-50 year were selected. Patients selected were subjected to three different scaling modalities namely hand scaling (control), hand scaling using sofscale (Experimental quadrant A) and ultrasonic scaling (Experimental quadrant B), in three different quadrants. Case report forms were used to document the tooth sensitivity, soft tissue pain after scaling, patient preference of instrumentation, ease of calculus removal, patient comfort, soft tissue irritation, time taken for scaling, Bleeding while scaling, pre and post operative sulcus bleeding index. In addition to the clinical criteria, the teeth treated were extracted and evaluated using the scanning electron microscope to show potential effects on cemntal surfaces. No difference in tooth sensitivity was appreciated between control and experimental quadrant A. There was a higher degree of tooth sensitivity when treated with ultrasonic. Patients in control group appreciated a higher degree of soft tissue pain. Hand scaling using softscale produced a lesser amount of pain and treatment with ultrasoincs was the least painful. Most of the patients preferred ultrasonic scaling (70%) Calculus removal was easier. Hand scaling using sofscale gel results in more patient comfort when compared to hand scaling alone. There was no significant difference in patient comfort between handscaling using sofscale and ultrasonic scaling. The percentage of reduction of sulcus bleeding index showed no difference between the 3 scaling modalities SEM evaluation revealed that there was no significant difference the 3 scaling modalities in relation to residual calculus, cleaning efficiency and damage to the root surface. This study concluded that treatment with sofscale gel appears to be safe and effective method for removal calculus as this did not damage cemental surfaces, nor did it cause any damage to soft tissue. "Your tratar is your calcified hate. Not only the microflora in your oral cavity but also your muddled thoughts, your obstinate squinting backward, the way you regree when you mean to progress, in other words, the tendency of your diseased gums to form germ catching pockets, all that, the sum of dental picture and psyche, betrays you, it is stored up violence, full of murdero us designs" Gunter Grass. PMID- 12765101 TI - Rebuilding a professional practice model. The return of role-based practice accountability. AB - There is no patient care without clinical practice. To improve the quality of health care, organizations must build a finely tuned and resilient clinical enterprise, one founded on clear role accountability and decision authority within the team. The author views scope of practice and professional standards as the foundation for practice accountability and decision authority. A case is made that an interdisciplinary, professional practice model is an appropriate delivery model in today's health care environment; a model that places the professional role in its rightful place as decision maker and supports the role's inherent accountability to evaluate and monitor practice performance. The importance of measuring professional practice performance is seen as a key link toward better understanding ways to reduce error and ensure patient safety. PMID- 12765102 TI - From survival to success. It takes more than theory. AB - Health care does not always attract change agents. As a result, norms, values, and rules become ingrained. Attempting to change this is difficult. This article will discuss how unique and personal skill sets of a leader complement elements of traditional change theory, resulting in moving organizations from minimal existence to national recognition. PMID- 12765103 TI - It takes a whole nation ... to create a health care system. AB - The biggest failure we face today is that we argue about the effects of poverty and the benefits of welfare rather than about how to sustain a reasonably cohesive and integrated community that optimizes human development for everybody. Yet that is exactly what must happen if we are to integrate our recreational, educational, health care, and health promotion systems and employment policies to create a healthier population. Care of the sick and the elderly is not a primary wealth- (or health-) producing social function; it is a derivative activity. If a society overinvests in it, it may underinvest in the real economy ... which then will not create the jobs, stability, and wealth that support the health of the population. This is not the kind of problem that the health care system alone, or any one sector of society, can actually tackle on its own. The challenge is to create real partnerships with all sectors: employers, labor leaders, educators, police, and governmental groups. PMID- 12765104 TI - Courage and today's nurse leader. AB - The virtue of courage is often overlooked in distinguishing successful leaders. This void is a reflection of the difficulty in defining just what courage is. Is courage facing risk without fear or overcoming fear to face risk? What are the differences between physical and moral courage? Can leaders develop courage? These and many other questions surround the nature of courage and how it pertains to leadership. It is the author's intent that readers have a general understanding of how courage affects nursing leadership in today's health care environment. PMID- 12765105 TI - Ethical climate in managed care organizations. AB - Managed care organizations employ nurses as medical utilization reviewers; however, little is known about the ethical climate of these organizations. This study describes different ethical climates in which utilization review nurses work and the implications of these differences for nurse administrators. The nurse participants, although demographically similar across three managed care organizations, perceived distinct ethical climates across the organizations. Nurses were employed to make complex decisions regarding medical care utilization; however, none of the organizations had an ethics committee to help nurse reviewers in this decision-making process. The need for such committees, as well as clarification of a consistent and deliberate ethical climate by nurse administrators, is discussed. PMID- 12765106 TI - Nurse leaders' perceptions of what compromises successful leadership in today's acute inpatient environment. AB - The purpose of this study was to gain an understanding of nurse leaders' perceptions of both the value of their roles in today's health care setting and their beliefs about how power and gender interface with role worth. Support for the theoretical significance of this research stemmed from Kanter's Structural Theory of Organizational Behavior. Four leaders were recruited at the executive level and 12 at the director/managerial level. The results of the deductive analysis supported Kanter's theory. Eighty-three percent of the nurse leaders validated that access to power, opportunity, information, and resources created an empowered environment, producing a climate that fostered leadership success and enhanced levels of job satisfaction among nurses. This study provided groundwork on the kinds of leadership traits that foster nursing satisfaction and on whether or not gender influences leadership effectiveness. The findings of this study are both timely and relevant for nurse leaders faced with the effects of the current supply-and-demand nursing shortage and with fiscal restraints mandated by managed care and regulatory agencies. PMID- 12765108 TI - Why are nurses leaving nursing? AB - The causes of nurses' exodus from acute health care delivery practice may lie more in intrinsic factors rather than the heretofore overtly expressed reasons. This article examines bureaucratic factors, issues related to the medical profession and medical/scientific discourse, and factors within the nursing profession itself that may contribute to a nurse's unhappiness and dissatisfaction that causes him or her to leave. Nursing as emotional work and the implications for the individual nurse, and nursing as moral and moral distress are discussed. Suggestions to facilitate retention are made for changing the work environment to feel valued for their skillfully applied humanness. PMID- 12765107 TI - The effects of intrapersonal, intragroup, and intergroup conflict on team performance effectiveness and work satisfaction. AB - Although numerous studies have focused on conflict management, few have considered the effects of unit technology and intrapersonal, intragroup, and intergroup conflict on team performance effectiveness and work satisfaction. The model was tested using a nonexperimental design. Path analysis using multiple regression was used to test the model. The nonrandom sample consisted of 141 nurses employed on 13 inpatient units at a state-supported, 597-bed academic medical center in a southeastern city. Findings indicated that intrapersonal conflict had a direct negative impact on intragroup conflict and work satisfaction. Intragroup conflict had direct negative effects on work satisfaction and team performance effectiveness. Unit technology had a direct negative impact on work satisfaction. Findings have implications for administrators to implement strategies to decrease a stressful work environment and increase team-building activities. PMID- 12765109 TI - Innovative educational strategies to prepare nurse executives for new leadership roles. AB - Distance education has grown over the last decades. As technology has improved, so has our ability to provide students with education options. Excelsior College has taken advantage of this digital age to bring together global nurse leaders and given them the opportunity participate self-paced learning at home. Excelsior is a leader in online education and now has expanded by offering masters courses with a specialty in clinical systems management and health care informatics to nurses worldwide. PMID- 12765110 TI - IT takes a village. Improving health care in the 21st century. PMID- 12765111 TI - Automated analysis of phase-contrast magnetic resonance images in the assessment of endothelium-dependent flow-mediated dilation. AB - Measurement of flow-mediated arterial dilation (FMAD) provides information regarding the status of peripheral arterial endothelial function. Although phase contrast magnetic resonance imaging (PC-MRI) can be used to measure FMAD, the manual analysis of one study (tracing regions of interest and processing data on 100 images) can require six or more hours. To enhance the clinical utility of the PC-MRI assessment of FMAD, we hypothesized that an automated technique (Multi Stage Intensity Thresholding or MSIT) for determining femoral arterial area and flow before and after cuff inflation over the thigh could be used to evaluate FMAD in a rapid, accurate, and reproducible manner. Compared with manual analysis, automated analysis detected a similar percentage change in peak FMAD between healthy individuals (17.2% vs 16.5%) and patients with congestive heart failure (4.0% vs 5.1%). The correlation between percentage changes in arterial area after cuff release derived manually and automatically was very good (r = 0.93). Analysis time for 100 images averaged 10 minutes with MSIT vs. 6 hours for manual analysis. In conclusion, rapid, accurate assessments of femoral artery FMAD can be obtained using Multi-Stage Intensity Thresholding. This methodology may prove useful for the rapid MRI assessment of peripheral arterial endothelial function in a clinical setting when studying patients with cardiovascular disease. PMID- 12765112 TI - Evaluation of left ventricular volumes and ejection fraction using fast steady state cine MR imaging: comparison with left ventricular angiography. AB - Previous studies demonstrated that magnetic resonance (MR) imaging consistently underestimated angiographic measurements of left ventricular (LV) volumes. The purpose of this study was to determine whether MR imaging with steady-state free precession acquisition (SSFP) can provide improved accuracy and reproducibility in measuring cardiac function in comparison with fast spoiled gradient echo cine MR imaging (SPGR). Twenty patients with cardiovascular diseases who underwent breath-hold cine MR imaging within one week of LV angiography were studied. Two sets of breath-hold cine MR images were obtained, one with SSFP and another with SPGR. The LV volumes determined by two breath-hold cine MR sequences were compared with the results by LV angiography. SPGR cine MR imaging consistently underestimated angiographic LV volumes. The mean difference of LV end-diastolic volume was -22.5 +/- 14.8 ml (p < 0.001) for short-axis planes and -27.7 +/- 21.5 ml (p < 0.001) for long-axis planes. In contrast, LV volumes measured by the SSFP imaging showed a good agreement with the results by angiography. The mean difference of LV end-diastolic volume was -2.5 +/- 14.3 ml (p = N.S.) for short axis planes and -10.9 +/- 15.1 ml (p < 0.01) for long-axis planes. Standard error of the estimation in measuring LV end-diastolic volume with the SSFP imaging was 3.9% for short-axis images and 4.9% for long-axis images. These values were 7.2% and 8.7% with the SPGR imaging. In conclusion, the SSFP acquisition can provide accurate and noninvasive assessments of LV volumes and ejection fraction within a reduced imaging time. PMID- 12765113 TI - Effects of missing dynamic images on myocardial perfusion reserve index calculation: comparison between an every heartbeat and an alternate heartbeat acquisition. AB - A commonly used method for analysis of first pass myocardial perfusion imaging is the calculation of a myocardial perfusion reserve index (MPRI) obtained by dividing the upslopes of the time-intensity curves at stress and rest. Perfusion data can be acquired with several different sequences with images acquired at every single, 2nd, or 3rd heartbeat. During data acquisition, some images of the dynamic series can be missed due to extra beats. Twenty-six patients underwent first-pass magnetic resonance perfusion imaging, acquiring images every heartbeat at rest and stress. The maximal upslopes of the myocardium and the left ventricle were calculated for the original image series and for the image series from which dynamics of every 2nd heartbeat were removed. Additionally for each of these situations the upslope calculations were repeated but with the removal of one or two dynamics during the maximal upslope. Images acquired every 2nd heartbeat yielded a lower upslope for the myocardium and the left ventricle, but the resulting MPRI was unchanged. Removing dynamics during the upslope resulted in a change of the MPRI by up to 44% for every heartbeat acquisition and by up to 56% for an alternate sampling. In conclusion, missing data points may affect the calculation of MPRI values and should be taken into account when using such values to define a threshold, which discriminates between normally and abnormally perfused myocardium. Furthermore, it may lead to false positive or negative results in individual cases. This effect is increased if data are acquired only every 2nd heartbeat. PMID- 12765114 TI - T1 mapping in patients with acute myocardial infarction. AB - Pixel-by-pixel calculation of T1 values (T1 mapping) has been used in different tissues to focus on T1 changes in a quantitative fashion. The aim of this study was to establish T1 mapping of human myocardium on a 1.5 Tesla system and to examine its diagnostic potential in patients with acute myocardial infarction (AMI). 8 patients with reperfused AMI (day 3 +/- 1) underwent multi-breath-hold MRI in a 1.5 Tesla system. Sets of five images with varying T1 weighting were acquired prior to and after the administration of contrast agent to generate images from calculated T1 values (T1 mapping). Prior to the contrast agent administration, all patients showed T1 prolongation in the area of infarction, which was identified in separate measurements using the delayed enhancement approach. Compared to noninfarcted areas, T1 values in the infarcted areas were increased by 18 +/- 7% (SE, p < 0.05). The spatial extent of the area of T1 prolongation was larger than that of the hyper-enhanced areas in conventional contrast-enhanced images. T1 maps obtained after the application of Gadolinium DTPA revealed a T1 reduction of 27 +/- 4% in infarcted tissue compared to noninfarcted areas (p < 0.05). The areas showing T1 reduction were in agreement with the hyper-enhanced regions in conventional T1-weighted images. T1 mapping visualizes changes in the longitudinal relaxation time induced by AMI. T1 mapping can detect myocardial necrosis without the use of contrast media. Information that can be extracted from a combination of pre- and postcontrast T1 maps exceeds that from conventional contrast studies. PMID- 12765115 TI - Myocardial scarring caused by left ventricular assist device (LVAD) insertion demonstrated by cardiovascular magnetic resonance. AB - We report three cases of dilated cardiomyopathy treated with left ventricular assist devices (LVAD), subsequently explanted. These mechanical devices are being increasingly used to support left ventricular function in the short and long term. We used cine and gadolinium-enhanced cardiovascular magnetic resonance (CMR) to examine the consequences of previous LVAD implantation. In all cases, there was apical akinesis and tethering on cine imaging. Early (< 5 minutes) imaging after gadolinium demonstrated apical hypo-enhancement, an avascular area of scar or thrombus, while late (> 10 minutes) imaging demonstrated transmural apical infarction which in one case extended into the inferior wall. The findings suggest that LVAD insertion may cause permanent myocardial fibrosis at the site of ventricular insertion, and the cases demonstrate the use of contrast-enhanced CMR in this scenario of iatrogenic ventricular scarring. PMID- 12765116 TI - Coronary magnetic resonance angiography: technical developments and clinical applications. AB - Magnetic resonance angiography (MRA) is a promising method for noninvasive coronary artery imaging. The interesting features of coronary MRA are its non invasive nature, the absence of ionizing radiation, and the possibility of imaging the coronary arteries without exogenous contrast. Moreover, three dimensional volume information and tomographic image planes in any direction can be obtained. The current review presents an overview of the recent technical developments of coronary MRA and the potential clinical applications. New MR techniques, such as parallel image acquisition, spiral imaging, steady-state free precession and the introduction of blood pool contrast agents may result in more efficient data acquisition. After implementation of these fast techniques, coronary MRA may be integrated in a comprehensive cardiac exam. PMID- 12765117 TI - Cardiac pacemakers, ICDs, and loop recorder: evaluation of translational attraction using conventional ("long-bore") and "short-bore" 1.5- and 3.0-Tesla MR systems. AB - PURPOSE: To evaluate magnet-related translational attraction for cardiac pacemakers, ICDs, and an insertable loop recorder in association with exposure to "long-bore" and "short-bore" 1.5- and 3.0-Tesla MR systems. MATERIALS AND METHODS: Fourteen cardiac pacemakers, four ICDs, and one insertable loop recorder were evaluated for translational attraction using deflection angle tests performed at the points of the highest spatial gradients for long-bore and short bore 1.5- and 3.0-Tesla MR systems according to ASTM guidelines. RESULTS: Deflection angles ranged from 9-90 degrees for the long-bore and from 11-90 degrees for the short-bore 1.5-T MR system. Deflection angles ranged from 23-90 degrees for the long-bore and from 34-90 degrees for the short-bore 3.0-T MR system. Three of the cardiovascular implants exhibited deflection angles > or = 45 degrees (i.e., indicating that they are potentially unsafe for patients) on the long-bore and short-bore 1.5-T MR systems. Eight implants exhibited deflection angles > or = 45 degrees on the long-bore 3.0-T MR system, while 14 exhibited deflection angles > or = 45 degrees on the short-bore 3.0-T MR system. In general, deflection angles for these cardiovascular implants were significantly (p < 0.01) higher on 1.5- and 3.0-Tesla short-bore compared to the long-bore MR systems. CONCLUSIONS: Several of the cardiovascular implants that underwent evaluation may be problematic for patients undergoing MR procedures using 1.5- and 3.0-T MR systems because of risks associated with magnet-related movements. Obviously, additional MR safety issues must also be considered for these implants. PMID- 12765119 TI - Comparison of MSCT and MRA in the evaluation of an anomalous right coronary artery. PMID- 12765118 TI - Detection of pseudoaneurysm of the left ventricle by fast imaging employing steady-state acquisition (FIESTA) magnetic resonance imaging. AB - This report highlights the importance of interpretating images throughout the course of a dobutamine MRI stress test. Upon review of the baseline images, the left ventricular (LV) endocardium was not well seen due to flow artifacts associated with low intracavitary blood-flow velocity resulting from a prior myocardial infarction. Physicians implemented a cine fast imaging employing steady-state acquisition (FIESTA) technique that was not subject to low flow artifact within the LV cavity. With heightened image clarity, physicians unexpectedly identified a LV pseudoaneurysm. PMID- 12765120 TI - [Virus infection in children after allogenic stem cell transplantation ]. AB - Allogenic hematopoietic cell transplantation (alloHCT) is the treatment of choice for various pediatric malignancies and nonmalignant diseases. The most prominent complication of allotransplantation is graft vs host disease (GvHD). The treatment of GvHD influence negatively function of immune system and increase risk of bacterial, fungal and viral infections. Clinical symptoms of viral infection may mimic GvHD and lead to inappropriate treatment. Human cytomegalovirus (CMV, Herpesviridae) has been recognized as most important viral pathogen after alloHCT. Increasing number of procedures, especially from alternative donors, requiring more intensive immunosuppression, led to identification more viral pathogens causing transplant related mortality and morbidity. Among them are adenoviruses (ADV, Adenoviridae), BK and JC viruses (Papovaviridae) and human herpes virus 6 (HHV-6, Herpesviridae). Frequency of complications caused by those pathogens is higher in children then in adults. PMID- 12765121 TI - [Regulation of NF-kappa B activity]. AB - Nuclear factor kappa B (NF-kappa B) is common proinflammatory transcription factor involved in expression of c.a. 300 different genes. It is usually present in the cytosol as an inactive complex and upon activation translocates into the nucleus. The mechanisms of activation of NF-kappa B are complex and they involve several different signaling pathways and plethora of proteins. In this minireview we describe the main deciphered as well as suggested mechanisms regulating of NF kappa B activity. PMID- 12765122 TI - [Biologic activity of lipopolysaccharides]. AB - Lipopolysaccharides (LPS, endotoxin) are known to be responsible for the initiation of endotoxic shock, therefore they can be targets for new preventive and therapeutic strategies. This review describes the biological and physicochemical properties of endotoxin--major component of the outer membrane of Gram-negative bacteria. The paper focuses on response of the host to endotoxin, recent knowledge about the target cells and receptors for LPS, mechanisms of innate immune response and cell signal transduction due to the LPS recognition. It also summarises results of studies on LPS structure influence on its biological activity. PMID- 12765124 TI - [Is use of nitric oxide (NO) synthase inhibitors justified for therapy of septic shock?]. AB - NO mediates many systemic reactions in humans and animals. It is produced by constitutive synthases (cNOS) and one inducible by cytokines and endotoxines (iNOS). We summarize, on the base of experimental and clinical data, the pathophysiological role of NO in septic shock as well as the effects of pharmacological application of NOS inhibitors. The analysed data show, that the contribution of NO to the pathomechanism of septic shock is rather heterogenous, therefore the clinical therapeutical application of selective iNOS inhibitors is impossible without adequate new monitoring tools. PMID- 12765123 TI - [Reconstitution of cyclophosphamide-induced, impaired function of the immune system in animal models]. AB - Cyclophosphamide (CY) is an alkylating agent used in chemotherapy of tumors and autoimmune disorders. The drug causes a large number of side-effects including deep, transient lymphopenia and neutropenia, thus rendering the immune system susceptible to infections. In this review we focus on the effects of CY on the haematopoetic system and the immune response in rodents. In addition, we present approaches aimed at reconstitution of lympho- and myelopoiesis using a spectrum of immunotropic factors including: thymic hormones, cytokines, low-molecular weight compounds, bacterial products and lactoferrin. PMID- 12765125 TI - [Molecular mechanisms regulating transport of low molecular weight substances in the hepatocyte]. AB - Transporters of sinusoidal and lateral membranes mediate uptake of inorganic and organic substances to hepatocyte. In hepatocyte lipophilic compounds are biotransformed (hydroxylated and conjugated) and excreted into bile and urine. Vectorial movement of solutes and water from blood to bile is maintained by pumps of ABC superfamily secreting bile salts (bile acid dependent flow) and other solutes: bilirubin, phospholipids, glutathione and inorganic salts (bile acid independent flow). PMID- 12765126 TI - HIV medicine after Barcelona conference: interview with Howard Grossman, M.D. Interview by John S. James. AB - We asked a leading HIV physician to summarize some of the treatment issues that clinicians are now looking at after the Barcelona conference. Topics in this interview include: Tenofovir, including first-line therapy; The shift from protease inhibitors to non-nucleoside RT inhibitors, for many but not all patients; Benefits of once-a-day regimens; Lipodystrophy, and strategies for avoiding or reducing it; Viral resistance testing; T-20; Prevention--including danger of superinfection with a new HIV strain. PMID- 12765127 TI - Pan-African AIDS treatment movement launched. AB - Activists from 21 African countries have launched a movement to organize the different sectors of societies, along with international institutions, to address the epidemic more successfully. PMID- 12765128 TI - Chinese activist detained by police, immediate action needed. AB - Dr. Wan Yanhai, widely considered one of China's leading AIDS activists, was detained by police in Beijing August 24. Amnesty International and AIDS organizations have called for immediate letters and email on his behalf. PMID- 12765129 TI - Cardiovascular diseases in people with HIV. PMID- 12765130 TI - HIV/AIDS in Asia and the Pacific. PMID- 12765131 TI - Mumbai meeting surveys AIDS in India. PMID- 12765133 TI - Bone disease & HIV. PMID- 12765132 TI - The HIV/AIDS drug pipeline. A status report. PMID- 12765134 TI - Treatment news from the XIV International AIDS Conference. PMID- 12765135 TI - Current issues in hepatitis C, HIV co-infection discussed at lunch and learn. PMID- 12765136 TI - HIV in women: an update. PMID- 12765137 TI - Neurosyphilis: how do you know, and what do you do? PMID- 12765147 TI - Arkansas Center update. HIV education in rural Arkansas. PMID- 12765148 TI - [Vitalii Iliich Popov (1896-1975)]. PMID- 12765149 TI - Retroviruses conference: Johns Hopkins report. AB - On March 1 the HIV newsletter from Johns Hopkins University published several short, focused reports on the Retroviruses conference, written primarily for physicians. These are available on the Web or by mail. We list most of the major topics covered. PMID- 12765150 TI - Obtaining the new HIV treatment guidelines. AB - How to order the new HIV/AIDS official adult treatment guidelines by mail if you do not have computer access; how to get your questions about treatments answered at the same office; and other guidelines available including prevention of opportunistic infections, and what to do in case of occupational exposure to HIV. In addition, the official Web site for the guidelines has a marked copy which shows where the current adult guidelines are changed from the previous version. PMID- 12765151 TI - Merck, Bristol-Myers Squibb announce major price reductions for poorest countries: major access progress but questions remain. AB - Two major pharmaceutical companies have announced new and much lower prices for antiretrovirals for African and some other poor countries, along with a uniform pricing policy which should simplify negotiations for access to the drugs. PMID- 12765152 TI - MSF (Doctors Without Borders) petition against South Africa lawsuit, deadline April 15. AB - Doctors Without Borders is collecting signatures from around the world, asking major pharmaceutical companies to drop the lawsuit which for three years has prevented South Africa from implementing it post-Apartheid reform of its medicines law. PMID- 12765153 TI - Southern Africa home care: conference report. AB - These Web reports of a recent conference in Gabarone, Botswana, provide an African view of the epidemic and home-care approaches now in use. PMID- 12765154 TI - [The 14th World Congress of Venous Surgeons: a scientific-practical forum at the turn of the century]. PMID- 12765155 TI - [6th International Congress of Pediatric Interventional Cardiologists (22-25 May 2002, Chicago, USA]. PMID- 12765156 TI - [Isaac Solomonovich Vengerovsky (1897-1967)]. PMID- 12765157 TI - [Ivan Demianovich Zhitniuk--eminent clinician and naval-military surgeon (100th anniversary of his birth)]. PMID- 12765158 TI - [Vladimir Mikhailovich Voskresensky (100th anniversary of his birth)]. PMID- 12765159 TI - New classes of HIV drugs on the horizon. A review of the presentation at the satellite symposium "New hope: advancing care in HIV infection" at the 15th annual Association of Nurses in AIDS Care conference, November 2002. AB - Despite the success of combination HAART in improving the clinical prognosis of HIV-infected patients, therapeutic options are limited for many patients, particularly those with extensive treatment experience. Resistance, cross resistance, and toxicity combine to threaten the durability of antiviral response, necessitating the development of novel agents. The HIV life cycle has many potential targets for inhibition in addition to the current reverse transcriptase and protease targets. These include the HIV integrase, nucleocapsid, and Tat proteins and the viral entry process. Entry inhibitors can be classified as attachment, coreceptor, and fusion inhibitors, according to the stage in the entry process at which they act. Most advanced in development of these is the fusion inhibitor enfuvirtide. Enfuvirtide is a potent inhibitor of HIV fusion whose novel mechanism of action and extracellular nature mean that it will induce limited cross-resistance to currently approved antiretrovirals and fewer toxicities. PMID- 12765160 TI - Clinical safety and efficacy of enfuvirtide (T-20), a new fusion inhibitor. A review of the presentation at the satellite symposium "New hope: advancing care in HIV infection" at the 15th annual Association of Nurses in AIDS Care conference, November 2002. AB - Proof of the concept that a fusion inhibitor could mediate clinically significant antiviral responses came from the demonstration that patients receiving 100 mg of enfuvirtide daily as monotherapy for 14 days achieved a mean reduction in viral load of 1.5 log10 copies/mL. Phase 2 studies established subcutaneous injection of 100 mg of enfuvirtide twice daily as the dosage of choice. In pivotal phase 3 trials, enfuvirtide was added to an optimized background regimen in heavily treatment-experienced patients. Virologic and immunologic responses were significantly better in patients receiving enfuvirtide in addition to optimized background regimens than in those receiving optimized background alone, with the incremental reduction in viral load between arms being 0.934 log10 copies/mL after 24 weeks of therapy. Subgroup analyses demonstrated this benefit to be similar regardless of age, sex, race, and baseline viral load. Enfuvirtide was well tolerated, with injection site reactions being the most common adverse event, although they were rarely treatment-limiting. PMID- 12765161 TI - Treatment with a new fusion inhibitor: patient issues with enfuvirtide (T-20). A review of the presentation at the satellite symposium "New hope: advancing care in HIV infection" at the 15th annual Association of Nurses in AIDS Care conference, November 2002. AB - The parenteral route of enfuvirtide administration requires that patients become familiar with reconstituting and administering the agent and in managing injection site reactions (ISRs). Since the majority of patients will experience at least one ISR, minimizing this effect is important, and several techniques can be taught to help alleviate the onset and symptoms of ISRs. Enfuvirtide patients usually encounter little difficulty with administration, and most report little impact of self-injections on their daily lives. However, instruction by nurses is crucial in training patients to administer enfuvirtide and to build this routine into their daily lives, thereby furthering the acceptance of enfuvirtide and promoting compliance. Patient education tools will provide clear and informative support, together with practical solutions to incorporate enfuvirtide therapy into daily life and travel. Easy access to the nurse and physician, coupled with an effective nurse-patient relationship, will be key in supporting good compliance with enfuvirtide therapy. PMID- 12765162 TI - Mercury and the risk of myocardial infarction. PMID- 12765163 TI - Survivors of the acute respiratory distress syndrome. PMID- 12765164 TI - Survivors of the acute respiratory distress syndrome. PMID- 12765165 TI - Mercury and the risk of myocardial infarction. PMID- 12765166 TI - Prevention of eclampsia. PMID- 12765167 TI - [Surgical management of acquired esophagorespiratory fistula: report of 2 cases]. PMID- 12765168 TI - Corticosteroid insufficiency in acutely ill patients. PMID- 12765169 TI - Corticosteroid insufficiency in acutely ill patients. PMID- 12765170 TI - Corticosteroid insufficiency in acutely ill patients. PMID- 12765171 TI - [Academician Dr. Hung Tao talking about BSE "crysis". Interview by Hung Tao]. PMID- 12765172 TI - 'My doctor said I had an abnormal chest X-ray'... PMID- 12765173 TI - Roy Hertz, M.D. (1909-2002): the cure of choriocarcinoma and its impact on the development of chemotherapy for cancer. AB - Dr. Roy Hertz is one of two scientists credited with discovering the first medical cure of a solid cancer. This paper presents a biographical history of Dr. Hertz and discusses his roles in the discovery of a cure for choriocarcinoma and as a pioneer for future research in cancer chemotherapy. This biography not only serves as a testament to the pioneering individuals in the field of chemotherapeutics but also represents the unique blend of medical, pharmacological, and physiological histories that led to the profound discovery. The timing and significance of the work of Drs. Hertz and Li cannot be overestimated. Their discovery was a spectacular success, demonstrating proof of the principle that chemotherapy can cure metastatic cancer and that an almost uniformly fatal cancer in young patients could be cured with a single chemotherapeutic agent, which stands as one of the greatest achievements in cancer research. PMID- 12765174 TI - Practicing evidence-based medicine: the Holy Grail? PMID- 12765175 TI - Evaluating the presence of antibodies to HPV as prognostic markers in cervical cancer patients. PMID- 12765178 TI - Interpretation of colposcopy data. PMID- 12765179 TI - Statistics to assess patient satisfaction with primary care called into question. PMID- 12765180 TI - Gingko: smart pill or not? PMID- 12765182 TI - Supporting scientists and research in Iraq. PMID- 12765183 TI - [The current status of the fundamental research in corneal disease in ophthalmology in China]. PMID- 12765184 TI - [Formats and categories of medical articles]. PMID- 12765185 TI - [Be of one mind to make the future]. PMID- 12765186 TI - [Pay attention to basic and clinical research on reconstruction of ocular superficiality]. PMID- 12765187 TI - The hot and the classic. PMID- 12765188 TI - [The 24th Report on Survey of the Adverse reaction to Radiopharmaceuticals (The 27th Survey in 2001)]. PMID- 12765189 TI - [Cover image. Bilateral nephroblastomatosis with secondary Wilms tumor manifestations]. PMID- 12765190 TI - Four new oleanane type saponins from Morina nepalensis var. alba. AB - Four new oleanane type saponins, monepalosides G-J (1-4), were isolated from the water-soluble part of the whole plant of Morina nepalensis var. alba Hand-Mazz. On the basis of chemical and spectroscopic evidence, their structures were determined as 3-O-alpha-L-arabinopyranosyl-(1 --> 3)-alpha-L-arabinopyranosyl oleanolic acid 28-O-beta-D-glucopyranosyl-(1 --> 6)-beta-D-glucopyranoside (monepaloside G, 1), 3-O-alpha-L-arabinopyranosyl-(1 --> 3)-beta-D-xylopyranosyl oleanolic acid 28-O-beta-D-glucopyranosyl-(1 --> 6)-beta-D-glucopyranoside (monepaloside H, 2), 3-O-alpha-L-arabinopyranosyl-(1 --> 3)-[beta-D glucopyranosyl-(1 --> 2)]-alpha-L-arabinopyranosyl oleanolic acid 28-O-beta-D glucopyranosyl-(1 --> 6)-beta-D-glucopyranoside (monepaloside I, 3), 3-O-beta-D glucopyranosyl-(1 --> 4)-beta-D-glucopyranosyl-(1 --> 3)]-alpha-L arabinopyranosyl oleanolic acid 28-O-beta-D-glucopyranosyl-(1 --> 6)-beta-D glucopyranoside (monepaloside J, 4), respectively. Two-dimensional NMR spectra, including H-H COSY, HMQC, 2D HMQC-TOCSY, HMBC and ROESY were utilized in the structure elucidation and complete assignments of 1H and 13C NMR spectra. PMID- 12765191 TI - A new biflavonoid from Ochna beddomei. AB - A new biflavonoid, 2,3-dihydroochnaflavone 7,4',7"-tri-O-methyl ether (1) together with two known biflavonoids namely, 2,3-dihydroochnaflavone (2) and ochnaflavone (3) were isolated from the stem bark of Ochna beddomei. The structures were determined by means of spectral and chemical studies. PMID- 12765192 TI - Three new saponins from the leaves of Ilex hylonoma. AB - Three new triterpenoid saponins, hylonosides III-V (1-3) have been isolated, along with three known oleanolic acid saponins (4-6), from the methanol extract of leaves of Ilex hylonoma. The structures were elucidated using a combination of homo- and hetero-nuclear 2D NMR techniques (COSY, TOCSY, NOESY, HMQC and HMBC) and negative FAB-MS. The new compounds were characterized as 3-O-beta-D glucopyranosyl-(1 --> 4)-beta-D-glucuronopyranosyl siaresinolic acid-28-O-beta-D glucopyranosyl ester (1), 3-O-beta-D-glucopyranosyl-(1 --> 4)-beta-D glucopyranosyl-(1 --> 2)-beta-D-glucuronopyranosyl siaresinolic acid-28-O-beta-D glucopyranosyl ester (2), 3-O-beta-D-glucopyranosyl-(1 --> 4)-beta-D glucopyranosyl-(1 --> 2)-beta-D-glucuronopyranosyl oleanolic acid-28-O-beta-D glucopyranosyl ester (3). PMID- 12765193 TI - Three new hecogenin glycosides from fermented leaves of Agave americana. AB - Eight steroidal compounds, including three new hecogenin glycosides, agamenosides D-F, were isolated from the fermented leaves of Agave americana. The structures of the new steroidal saponins were elucidated by spectroscopic data and chemical methods. The activity of the isolated compounds on deformations of mycelia germinated from conidia of Pyricularia oaryzae P-2b was evaluated. PMID- 12765194 TI - Colorimetric estimation of total iridoid content of Picrorhiza kurrooa. AB - A colorimetric method has been developed for the selective analysis of the total iridoid content of the rhizomes of Picrorhiza kurrooa in terms of catalpol. The method of analysis is based on the reaction between iridoid compounds and primary amine. The iridoid glycosides present in P. kurrooa are mainly the esters of catalpol, and can be easily converted into catalpol by saponification. Catalpol thus obtained by hydrolysis is treated with glycine in acidic medium to give a purple color with a maxmimum absorption of 542 nm. The method was validated as per the ICH guidelines for linearity, accuracy and precision. Several other rhizome samples of the plant were also assayed using this method. The method developed is precise, sensitive, reproducible and easy to perform and can be used for the standardization of crude drug. PMID- 12765195 TI - Stilbenoids from the lianas of Gnetum pendulum. AB - The first new isorhapontigenin dimer glucoside gnetupendin D (1) and a stilbene dimer coupled by a resveratrol and an oxyresveratrol with a cis-dihydrobenzofuran were obtained from the lianas of Gnetum pendulum C. Y. Cheng (Gnetaceae), along with nine known compounds. Their structures and stereochemistry were elucidated by chemical and spectral evidence, especially 2D NMR spectroscopic techniques. Pharmacological activities on anti-inflammation have been tested. PMID- 12765196 TI - Anti-invasion and anti-angiogenesis effect of taxol and camptothecin on melanoma cells. AB - Two highly invasive melanoma cell lines B16BL6 and B16F10 were used to investigate the anti-invasion and antiangiogenesis action of taxol and camptothecin (CPT). The adhesion of melanoma cells was tested by optical absorbance at 545 nm. The invasive activity of these cells was tested in a transwell chamber assay. The cell migration within a 3D collagen matrix was recorded with a time-lapse video recorder and analyzed by computer-assisted cell tracking. Gelatin zymography was used to study the metalloproteinase activity. The chicken chorioallantoic membrane (CAM) model was used to study the anti angiogenesis effect of the two drugs. The results demonstrated that both taxol and CPT could inhibit the migration of B16F10 cells, and inhibit the adhesion of B16F10 to fibronectin and laminin. They can reduce the metalloproteinase secretion of HT1080 and exhibit the antiangiogenesis effect in the CAM model. Taxol showed a highly anti-invasion effect on B16BL6 cells while CPT did not exhibit such an effect. PMID- 12765197 TI - C-glucoside flavonoids from the leaves of Crataegus pinnatifida Bge. var. major N.E.Br. AB - Two new acetyl C-glucoside flavonoids, 8-C-beta-D-(2"-O acetyl)glucofuranosylapigenin and 3"-O-acetylvitexin, along with four known C glucoside flavonoids, vitexin, 6"-O-acetylvitexin, 2-O-acetylvitexin, and 2"-O rhamnosylvitexin were isolated from the leaves of Crataegus pinnatifida Bge. var. major N.E.Br. Their structures were elucidated by spectroscopic means and chemical evidence. PMID- 12765198 TI - Terminoside A, a new triterpene glycoside from the bark of Terminalia arjuna inhibits nitric oxide production in murine macrophages. AB - Terminoside A (1), a new oleanane-type triterpene was isolated from the acetone fraction of the ethanolic extract of stem bark of Terminalia arjuna. The structure was established as olean-1alpha,3beta,22beta-triol-12-en-28-oic acid 3beta-D-glucopyranoside. On the basis of spectral data and chemical reactions, terminoside A, potently inhibited nitric oxide (NO) production and decreased inducible nitric oxide synthase (iNOS) levels in lipopolysaccharide-stimulated macrophages. PMID- 12765199 TI - Chemical constituents of Pyrrosia petiolosa. AB - The new flavone diglycoside 7-O-[6-O-(alpha-L-arabifuranosyl)-beta-D glucopyranosyl]-gossypetin, named as pyrropetioside, along with 13 known compounds (including 3 artifacts) have been isolated from Pyrrosia petiolosa. Their structures have been elucidated by means of chemical and spectroscopic methods including IR, MS, 1D and 2D NMR techniques. PMID- 12765200 TI - Suppression of IL-8 gene transcription by resveratrol in phorbol ester treated human monocytic cells. AB - Resveratrol (3,4',5-trihydroxy-trans-silbene), a natural phytoalexin found in grapes and other food products, has promising anti-inflammatory and anticancer effects. To observe the modulation of interleukin-8 (IL-8) production in human monocytic cells by resveratrol and explore its mechanism at the gene transcription level, U937 cells were stimulated with phorbol 12-myristate 13 acetate (PMA) for 24h. IL-8 protein in supernatants was measured by radioimmunoassay. The cytotoxicity of PMA, dexamethasone and resveratrol was accessed by MTT cell proliferation assay. The RNA level of glyceraldehyde 3 phosphate dehydrogenase (GAPDH) and IL-8 were detected by RT-PCR using specific primers. DNA binding activities of NF-kappaB and AP-1 were examined by electrophoretic mobility shift assay (EMSA). 0.01-100 nM PMA could significantly induce IL-8 production in U937 cells; 10 microM Dexamethasone and 10, 1, 0.1 microM resveratrol could inhibit PMA-induced IL-8 protein production and mRNA accumulation. The cytotoxicity did not contribute to their inhibitory effect. The DNA binding activity of AP-1 was inhibited by dexamethasone and resveratrol, but resveratrol has little effect on PMA-induced NF-kappaB activation. Resveratrol could inhibit PMA-induced IL-8 production in U937 cells at protein and mRNA levels. The suppression of IL-8 gene transcription by resveratrol was, at least partly, due to inhibition of AP-1 activation. PMID- 12765201 TI - Self-reported asthma prevalence and control among adults--United States, 2001. AB - Asthma is a chronic illness that has been increasing in prevalence in the United States since 1980. In 2000, asthma accounted for 4,487 deaths, approximately 465,000 hospitalizations, an estimated 1.8 million emergency department (ED) visits, and approximately 10.4 million physician office visits among persons of all ages. To provide prevalence data for state and local health department asthma programs, the Behavioral Risk Factor Surveillance System (BRFSS) collects data each year from the 50 states, the District of Columbia, and three U.S. territories. This report summarizes asthma prevalence data for adults collected from the 2001 BRFSS survey and from the eight states that used the adult asthma history module. Findings from BRFSS indicate that approximately 7.2% of U.S. adults have current asthma. ED visits for asthma varied more than any other characteristic among the eight states that used the adult asthma history module. In Mississippi, 67.3% of respondents with current asthma reported no ED visits during the preceding 12 months, compared with 87.6% in Washington state. Continued use of the BRFSS asthma prevalence questions and the asthma history module will allow state health departments to monitor trends in asthma prevalence and control and to direct public health asthma interventions. PMID- 12765203 TI - Update: Severe acute respiratory syndrome--United States, 2003. AB - CDC continues to work with state and local health departments, the World Health Organization (WHO), and other partners to investigate cases of severe acute respiratory syndrome (SARS). During November 1, 2002-April 30, 2003, a total of 5,663 SARS cases were reported to WHO from 26 countries, including the United States; 372 deaths (case-fatality proportion: 6.6%) have been reported. This report updates information on reported SARS cases among U.S. residents and provides an overview regarding CDC's issuance of travel alerts and advisories. PMID- 12765202 TI - Women with smallpox vaccine exposure during pregnancy reported to the National Smallpox Vaccine in Pregnancy Registry--United States, 2003. AB - In the absence of circulating smallpox, pregnant women should not be exposed to live vaccinia virus contained in the smallpox vaccine. The smallpox vaccine should not be administered to women who are pregnant or might become pregnant within 4 weeks after vaccination because of the risk for fetal vaccinia, a rare but serious infection of the fetus. In addition, persons who have close contact (e.g., household contact or sexual contact) with pregnant women are advised to forego vaccination. To prevent inadvertent exposure of pregnant women to vaccinia virus, screening for pregnancy is a component of pre-event smallpox vaccination programs. To monitor outcomes of pregnancy in women exposed to smallpox vaccines, CDC, in collaboration with the Department of Defense (DoD) and the Food and Drug Administration (FDA), has established the National Smallpox Vaccine in Pregnancy Registry. This report summarizes data from the registry about these exposures. CDC and the Advisory Committee on Immunization Practices (ACIP) recommendations to screen for pregnancy as a contraindication to smallpox vaccination appear to be effective at preventing inadvertent exposures. PMID- 12765204 TI - Updated interim surveillance case definition for severe acute respiratory syndrome (SARS)--United States, April 29, 2003. AB - CDC's interim surveillance case definition for severe acute respiratory syndrome (SARS) has been updated to include laboratory criteria for evidence of infection with the SARS-associated coronavirus (SARS-CoV) (Figure, Box). In addition, clinical criteria have been revised to reflect the possible spectrum of respiratory illness associated with SARS-CoV. Epidemiologic criteria have been retained. The majority of U.S. cases of SARS continue to be associated with travel, with only limited secondary spread to household members or health-care providers. PMID- 12765205 TI - Effects of gender and level of parental involvement among parents in drug treatment. AB - Most studies of parents in drug treatment have focused exclusively on mothers, and few studies have examined the effects of parents' level of involvement with their children on the parents' drug use and psychological functioning, either before or after treatment. This study examined mothers and fathers (n = 331) who were parents of children under the age of 18; participants were sampled from 19 drug treatment programs across four types of treatment modalities in Los Angeles County. A majority of each group (57% of 214 mothers and 51% of 117 fathers) were classified as being highly involved with their children. At the baseline assessment, higher parental involvement was related to lower levels of addiction severity, psychological severity, and symptoms of psychological distress, and to higher levels of self-esteem and perception of parenting skills. In general, fathers had higher levels of alcohol and drug-use severity than did mothers, but fathers who were more involved with their children showed lower levels of addiction severity than fathers who were less involved. Parental involvement at baseline was unrelated to drug use at the 12-month follow-up, although parents who were less involved with their children reported experiencing more stressors. Given the association of parental involvement with lower levels of addiction severity and psychological distress at baseline, treatment protocols should build upon the positive relationships of parents with their children, and seek to improve those of less-involved parents. PMID- 12765206 TI - A comparison between dually diagnosed inpatients with and without Axis II comorbidity and the relationship to treatment outcome. AB - The presence of a personality disorder (PD) has been associated with certain types of poor treatment outcomes in patients with substance use disorders (SUDs). The purpose of this study was to determine the prevalence of comorbid PDs in psychiatrically hospitalized adults with both non-SUD Axis I disorders and SUDs, and to assess the relationship between Axis II psychopathology and degree of pretreatment addiction severity and treatment outcome. METHOD: One hundred consecutive inpatients admitted to a mixed dual diagnosis inpatient unit were assessed using semistructured interviews for SUDs, non-SUD Axis I disorders, and PDs. Pretreatment severity was assessed using a modified version of the Addiction Severity Index (ASI). Outcome measures were assessed both during hospitalization and at an initial follow-up appointment after discharge. Statistical analyses were performed comparing dually diagnosed patients with and without Axis II psychopathology. RESULTS: A significant number (53%) of the patients met criteria for at least one personality disorder. Of the PDs, Cluster B PDs were the most prevalent, particularly borderline personality disorder (74%) and antisocial personality disorder (66%). Dually diagnosed patients without an Axis II diagnosis had less severe pretreatment severity measures. During hospitalization, patients with Axis II disorders had higher levels of psychopathology on the Brief Symptom Inventory (BSI) subscales of sensitivity and hostility. However, there was no difference in overall degree of global improvement during hospitalization. During follow-up, patients with Axis II disorders were significantly less likely to be compliant in attending their initial follow-up appointment. CONCLUSIONS: Dual diagnosis inpatients with PDs appear to improve as much as patients without PDs during their inpatient hospitalizations; however, they appear to be less likely to be compliant with attending their initial follow-up appointment. PMID- 12765207 TI - Differential effects in humans after repeated administrations of zolpidem and triazolam. AB - Zolpidem, a commonly prescribed hypnotic, is an imidazopyridine that purportedly has a unique benzodiazepine-receptor binding profile. Despite its unique binding profile, human laboratory experiments have generally failed to demonstrate meaningful behavioral pharmacological differences between zolpidem and classic benzodiazepine-receptor agonists like triazolam. In this article, two groups of nondrug-abusing volunteers received 15 mg zolpidem (N = 11) or 0.375 mg zolpidem (N = 15) on four separate occasions and placebo on two other occasions. In both groups, the order of drug administration was quasi-random. Drug effects were assessed with a battery of laboratory performance tasks and subject-rated drug effect questionnaires. Zolpidem and triazolam produced prototypical sedative-like effects (e.g., impaired performance, increased subject-ratings of sedation). The performance-impairing effects of triazolam, but not zolpidem, were significantly less after the final administration relative to the initial administration. The subject-rated effects of both zolpidem and triazolam were significantly less after the final administration relative to the initial administration. The results of this experiment suggest that zolpidem and triazolam differ in terms of tolerance-producing effects. Future studies should assess the tolerance-producing effects of zolpidem across a range of doses, as well as other novel sedatives such as zaleplon. PMID- 12765210 TI - Retention of court-referred youths in residential treatment programs: client characteristics and treatment process effects. AB - The juvenile justice system relies heavily on residential treatment services for adolescents. Because treatment dropout limits the likely effectiveness of these services, in this study we examine the client and program characteristics associated with program retention among a sample of adolescent probationers referred to residential rehabilitation by the Juvenile Court in Los Angeles. Participants in the present study (n = 291) are a subset of those in the Adolescent Outcomes Project, conducted within RAND's Drug Policy Research Center, to examine the outcomes of youths entering treatment at seven residential treatment programs. Three months after a preadmission interview, youths were asked about their perceptions of counselors at the program, other residents, and their feelings of safety in the program. In addition, they were asked whether they needed and had received various services (e.g., job training, legal advice, family counseling). Results of a multivariate survival analysis revealed that pretreatment characteristics including motivation and substance use severity, as well as treatment program factors including safety, and perceived over- and underprovision of services, contribute significantly to the prediction of retention. Pretreatment environmental risk factors and ratings of program counselor and resident support were marginally significant. These results imply that changes in adolescent residential program delivery may serve to increase retention rates, thus improving long-term outcomes. PMID- 12765209 TI - Quality of life assessments by adult substance abusers receiving publicly funded treatment in Massachusetts. AB - PURPOSE: Quality of life (QOL) has become an important endpoint in clinical trials and studies of medical interventions for many chronic diseases, but has not been widely studied in the substance abuse field. The purpose of this study was to measure QOL in a large sample of adult substance abusers (SA), to compare QOL ratings with those for other chronic diseases, and to assess factors influencing QOL levels in this population. METHODS: Hour-long personal interviews were administered to 570 randomly selected substance abuse clients in six detoxification centers and seven outpatient facilities in Massachusetts. Two QOL instruments--the Multidimensional Index of Life Quality (MILQ) and the SF-12- were used to measure QOL. Substance use problems were measured by the Addiction Severity Index Drug and Alcohol composite scores. QOL scores were compared to those for patients with other chronic diseases. Multivariable regression analysis was used to estimate the influence of demographic characteristics, co morbidities, and substance use on QOL summary scores. RESULTS: SA clients reported significantly lower QOL scores than patients about to undergo heart surgery on seven of the nine MILQ domains as well as the MILQ QOL summary index. SA clients also had much lower mean scores than the general population on both the SF-12's Mental Health (ES = -1.80) and Physical Health (ES = -0.58) component scales. SF-12 physical component scores were similar to those for other chronic diseases, but mental component scores were markedly lower and comparable on average to clinically depressed groups. In the regression model, MILQ Index scores were significantly lower for dual diagnosis and detoxification cases. Index scores were negatively associated with ASI drug composite scores, but not with ASI alcohol scores. IMPLICATIONS: The results of this study suggest that the physical functioning of adult substance abusers is similar to the levels for patients diagnosed with other serious chronic diseases, but that mental functioning is much lower. QOL provides information about functioning and well being that is not captured by traditional measures of substance use, and may soon begin to play a more prominent role in evaluating the effectiveness of treatment services for substance abusers. PMID- 12765208 TI - Mediators of effectiveness in dual-focus self-help groups. AB - Although research on the effectiveness of 12-step group participation has been increasing, there has been little examination of the processes by which such participation leads to positive outcomes. Two kinds of factors have been proposed as mediating between 12-step group affiliation and outcomes for members, common process factors that have been identified in a range of behavioral treatments and factors that are relatively unique to the 12-step model. The study tested the hypotheses that two common process factors (internal locus of control and sociability) and two unique factors (spirituality and installation of hope) mediate the effects of 12-step group affiliation on drug/alcohol abstinence and health promoting behavior. The study respondents were members of a dual focus 12 step-based fellowship, Double Trouble in Recovery (DTR), designed to address issues of both substance use and mental health. Members of 24 DTR groups in New York City were recruited and followed-up for 1 year. The degree of 12-step group affiliation during the study period was associated with more positive outcomes at follow-up. Internal locus of control and sociability mediated the effects of 12 step group affiliation on both outcomes, whereas spirituality and hope acted as mediators only for health promoting behavior. Understanding that the therapeutic factors inherent in 12-step are not mysterious, but appear to capitalize on well documented social learning principles, may increase the acceptance of 12-step programs among addiction and mental health professionals. PMID- 12765211 TI - Comparing treatments for dual diagnosis: twelve-step and self-management and recovery training. AB - The purpose of this study was to compare the effectiveness of 12-step and cognitive-behavioral (Self-Management and Recovery Training [SMART]) approaches for persons with a dual diagnosis of serious mental illness and substance use disorder in an intensive outpatient/partial hospitalization setting. Participants (n = 112) were alternately assigned to the two treatment conditions, with 50 participants completing the 6-month treatment program. Assessments occurred at baseline, 3 months, and 6 months during treatment, and at 3- and 12-month follow ups. Analyses were conducted on participants who had completed 3 months of treatment (n = 70). The 12-step intervention was more effective in decreasing alcohol use and increasing social interactions. However, a worsening of medical problems, health status, employment status, and psychiatric hospitalization were associated with the 12-step intervention. SMART was more effective in improving health and employment status, but marijuana use was greater for SMART participants. Improvements in alcohol use and life satisfaction occurred in both approaches. Covariates associated with treatment outcome were identified, with greater attendance being positively related to outcome. Involvement with the criminal justice system was positively related to treatment completion but negatively associated with medical problems. Less alcohol use, fewer medical problems, and better financial well-being at baseline were associated with better attendance. PMID- 12765212 TI - Is self-mastery always a helpful resource? Coping wiith paradoxical findings in relation to optimism and abstinence self-efficacy. AB - This study investigated the relationship between three personal resources (self mastery, optimism, and abstinence self-efficacy) and ways of coping among recovering substance abusers (n = 52) residing in Oxford Houses. Although residents' scores on optimism and abstinence self-efficacy were significantly correlated in a positive direction, residents' self-mastery scores were significantly and negatively related to both optimism and abstinence self efficacy scores. However, residents reported using significantly more emotion focused than problem-focused coping even though there was a significant positive relationship between emotion-focused coping and self-mastery. These paradoxical findings might be due to communal living and 12-step philosophy that is antithetical to a sense of control, which may have facilitated residents' emotional regulation to stress. Furthermore, significant relationships between coping strategies and personal resources imply that active coping strategies are instrumental in recovering substance abusers' use of optimism and abstinence self efficacy, whereas passive coping strategies are involved with self-mastery. In addition, African American residents reported using significantly more wishful thinking coping, suggesting ethnicity might be an important aspect of the coping process. Overall, findings suggest that optimism and abstinence self-efficacy are effective personal resources for recovering substance abusers in 12-step programs. PMID- 12765213 TI - Use and assessment of complementary and alternative therapies by intravenous drug users. AB - Intravenous drug users often have many health conditions in addition to their drug addiction, yet may be isolated from conventional sources of care. They have never before been examined for their use of complementary and alternative medicine (CAM) therapies. Our purpose was to study the prevalence and predictors of CAM use among persons with a history of intravenous drug use through a cross sectional survey of intravenous drug users examining their utilization of health services, including CAM therapies. A total of 548 persons with a history of intravenous drug use, recruited from a needle-exchange program and a methadone maintenance clinic, both in Providence, Rhode Island, participated. Overall prevalence of any CAM use in the past 6 months, frequency of use of individual named CAM therapies and domains, and demographic and clinical characteristics associated with CAM users, reasons for CAM use and self-perceived effectiveness of CAM were also measured. Of the 548 participants, 45% reported use of at least one CAM therapy. The top three therapies--religious healing, relaxation techniques, and meditation--were all from the mind-body domain. Having a higher education and lower self-rated health were the two strongest predictors of CAM use, followed by having a regular doctor or clinic, being white and younger. There was a high level of self-perceived effectiveness of CAM therapies (4.1 on a scale of 1-5), and CAM users were likely to use CAM for reasons related to their addiction. PMID- 12765214 TI - Assessing sex differences on treatment effectiveness from the drug abuse treatment outcome study (DATOS). AB - Men and women entering drug treatment programs are known to differ in demographic characteristics and psychosocial behavioral patterns. To be effective, any such program that caters to individuals from both sexes should identify and address these gender-based variations. Studies have also reported clinical differences in the effect of drugs on men and women addicts. Here, we examine whether the treatment is equally effective on men and women, when several demographic covariates are controlled. We construct a "problem severity index" to categorize individuals based on how acute their problems were at the start of the program. We also examine how the choice of treatment modality affects treatment success. Cumulative logit models were used in our analysis. The choice of treatment modality is sex specific, although sex did not appear to be a significant factor for treatment effectiveness when we controlled for other explanatory variables. PMID- 12765215 TI - Searching for universals: preliminary evidence for the validity of substance abuse subtypes in a sample of Mexican American youths. AB - Recent studies of substance abuse typologies indicate that multivariate models originally developed for identifying subtypes of alcoholics are valid among users of other substances, including heroin, cocaine, and marijuana. Little is known regarding the generalizability of these subtypes across culturally different subgroups. If substance abuse typologies are universal, then controlling for subtype may prove to be an effective research strategy in studies attempting to distinguish the separate and specific contribution of culturally related factors to risk for substance use and to treatment response. In this article, we present data that replicate the Type A-Type B distinction, developed by Babor et al., in a sample of Hispanic youths with substance involved problems. PMID- 12765216 TI - Childhood trauma and adult prostitution behavior in a multiethnic heterosexual drug-using population. AB - A cross-sectional study of the association between severity of childhood trauma and adult prostitution behaviors was conducted among 676 heterosexual drug addicts in San Antonio, Texas. Three hundred and fifty eight women and 338 men taking part in a national multisite program for AIDS prevention research completed the Childhood Trauma Questionnaire as part of a comprehensive risk behavior assessment. Women addicts in the sample were less educated, more likely to be in a common-law relationship, living with someone of the opposite sex or separated, and had lower incomes in comparison to men addicts. Among male subjects,higher educational levels and older age were positively associated with prostitution activities. Single female subjects were three times more likely to engage in selling sex than married subjects. Single women with higher incomes were more likely to be prostituting than single women with lower incomes. Black women reporting severe degrees of emotional abuse, emotional neglect, or physical neglect were more likely to engage in prostitution behavior than Hispanic or white women with similar levels of trauma. Black men with a history of childhood physical abuse were more likely to use prostitutes than Hispanic or white men. PMID- 12765217 TI - Two cases of severe gamma-hydroxybutyrate withdrawal delirium on a psychiatric unit: recommendations for management. AB - Many psychiatric professionals are unfamiliar with gamma-hydroxybutyrate (GHB), an increasingly popular drug of abuse. GHB withdrawal can lead to psychosis and agitation, and patients may present to psychiatric facilities for treatment. Withdrawal may progress to delirium, with the potential for severe or even fatal medical complications. Therefore, it is imperative for psychiatric professionals to understand how to treat these patients. In this article, we describe two cases of severe GHB withdrawal syndrome that were treated in our inpatient psychiatric unit. These are among the most severe cases reported. Pertinent literature is reviewed and suggestions for treatment are discussed. PMID- 12765218 TI - Differential, histochemical and immunohistochemical changes in rat hepatocytes after isoflurane or sevoflurane exposure. AB - Differential, histochemical and immunohistochemical changes were observed in hepatocytes from immediately to 7 days after isoflurane or sevoflurane exposure (at H 0 to on Day 7) to study the process of development and recovery in anesthetic-induced hepatic injury. A total of 570 7-week-old male Sprague-Dawley rats with or without phenobarbital treatment were exposed to isoflurane or sevoflurane in 100%, 21%, or 10% oxygen, or to 10% oxygen alone for 2h. In phenobarbital-treated rats, hepatocytes both with and without anesthetic exposure markedly changed in 10% oxygen at H 0. Glycogen and ribosomal ribonucleic acid (rRNA) disappeared at H 0 and at H 6, respectively, and at H 6, AST levels in the blood rose. From H 6 to Day 1, necrosis developed more markedly and widely in zone 3 hepatocytes exposed to anesthetics in 10% oxygen than in those exposed to oxygen alone. All degenerated tissues had returned to normal levels by day 7. Recovery of the hepatolobular structure may be attributed to rearrangement of remaining hepatocytes in the portal vein area. Both the disappearance of glycogen and rRNA and the increase in blood AST levels after exposure to isoflurane or sevoflurane are considered to be factors contributing to the induction of necrosis around the central vein. The grade of isoflurane-induced hepatic injury was found to be significantly higher than that of sevoflurane. PMID- 12765219 TI - Protective effect of leukotriene receptor antagonist montelukast on smoking induced lung injury in Wistar rats. AB - Increased activation of alveolar macrophage, neutrophil and mast cell has been proven in cigarette smoking (CS)-related lung disorders (CSLD). An increased production of cysteinyl-leukotrienes (LTs), which are mediators secreted from the mentioned cells, in response to CS has been shown in humans. The protective effect of LT1 receptor-1 antagonist (LTR-1AT) on CSLD is, however, not known. In this study we aimed to determine whether there is any protective effect of a LTR 1AT, montelukast (MK), on CSLD in Wistar rats. Nine controls and twenty-three smoke-exposed rats were enrolled into this study. Controls were exposed to non filtered air, and the smoke-exposed rats were exposed to CS for 6 h/day, 6 days/week for three weeks. The CS-exposed rats were also treated with 0.1 mg/kg/day of MK or saline. Morphometric criteria for lung injury were determined as the mean linear intercept of alveolar septa (Lm), the volume density of alveolar septa (Vvspt) and the density of the alveolar surface area per unit volume of lung parenchyma (Sva.pa). Lung mast cells (LMC), which are a major source of LTs, were also counted. Results showed that Lm of the control group was significantly lower and Vvspt, Sva.pa of the controls were significantly higher compared to those of the CS-exposed groups. Animals treated with MK had significant protection against CSLD. Lm was significantly higher and Vvspt, Sva.pa were lower in the saline group than in the MK-treated group. The number of LMC in the CS-exposed groups was also significantly higher than that in the control group. Based on these results, one can suggest that some part of the pathogenesis of CSLD may be related to an enhanced LTs synthesis and LTR-1AT. Therefore, montelukast may protect against active or passive smoking-induced lung injury and related disorders. PMID- 12765220 TI - Determination of etoposide serum concentrations in small pediatric samples by an improved method of reversed-phase high-performance liquid chromatography. AB - Several specific assays have been developed for the measurement of etoposide in biological fluids. As large samples are required for high sensitivity, these systems are not appropriate for a pediatric practice. In the present study, however, an improved method for the determination of serum levels of the anticancer drug etoposide was developed, using high-performance liquid chromatography with fixed-wavelength ultraviolet detection. Etoposide was extracted from serum using dichloromethane. The efficiency of extraction from serum was 85.7 +/- 7.7% for etoposide and 81.1 +/- 8.4% for diphenylhydantoin, the internal standard. The serum concentrations of etoposide were measured in 0.2 ml serum samples. The lower limit of detection was 50 ng/ml. Each measurement was completed within 5 min. The linear quantitation range for etoposide was 0.05-50 microg/ml. This assay presents an alternative method for routine measurement of serum levels of etoposide in the pediatric oncology setting. PMID- 12765221 TI - Prediction markers for respiratory distress syndrome: evaluation of the stable microbubble test, surfactant protein-A and hepatocyte growth factor levels in amniotic fluid. AB - Surfactant treatment in infants with respiratory distress syndrome (RDS) has decreased neonatal mortality. With the advent of this therapy, it has become important to predict accurately the fetal lung maturity of a fetus before delivery. We evaluated the stable microbubble test (SMT), surfactant protein-A (SP-A) and hepatocyte growth factor (HGF) in amniotic fluid as predicting markers for RDS. Of 55 amniotic fluid samples obtained by amniocentesis from women less than 37 weeks pregnant, the SMT values were as follows: sensitivity 76.5%, specificity 84.2%, positive predictive value 68.4%, negative predictive value 88.9% and overall accuracy 81.8%. For SP-A, the values were 88.2%, 65.8%, 53.6%, 92.6% and 72.7%, respectively. If we used both SMT and SP-A, we could diagnose with 100% accuracy that a case with measurements of SMT > or = 2 and SP-A > or = 420 ng/ml would not complicate with RDS (24/24). However, the RDS diagnostic accuracy of HGF does not equal to those of SMT and SP-A levels. We concluded that the rapidity, simplicity and reliability of SMT was very useful during 24-36 weeks of gestation as a bedside procedure to predict fetuses likely to develop RDS. We also noted the additive effect of SP-A in improving the accuracy of lung maturity diagnosis. PMID- 12765222 TI - Myofibroblasts proliferation of idiopathic and collagen vascular disorders associated nonspecific interstitial pneumonia. AB - Nonspecific interstitial pneumonia (NSIP) has been recognized as a separate histological classification of interstitial lung disease. Similar features are found not only in idiopathic NSIP, but also in NSIP associated with collagen vascular disorder (CVD-NSIP). We examined the clinical symptoms, laboratory findings, and prognosis of 13 cases of idiopathic NSIP and 11 cases of CVD-NSIP. Immunohistochemical staining was performed using the streptavidin/biotin/peroxidase method with anti-alpha-smooth muscle actin antibody. No differences in the distribution of clinical features, laboratory findings, and prognosis were observed between idiopathic NSIP and CVD-NSIP. In immunohistochemical staining of the fibrosing areas, myofibroblasts were observed in 7 of 13 idiopathic NSIP cases, but in 10 of 11 CVD-NSIP cases. With regards to intra-alveolar organization, myofibroblasts were observed in all 10 CVD-NSIP cases, but they were observed in only 2 of 9 idiopathic NSIP cases. We found a significantly higher myofibroblast proliferation in the intra-alveolar organization of CVD-NSIP compared to idiopathic NSIP. Clinically, idiopathic NSIP and CVD-NSIP are similar, but are pathologically different. PMID- 12765223 TI - Prognostic factors in children with purulent meningitis in Turkey. AB - In this study the clinical and laboratory findings of 48 children with purulent meningitis were examined, prospectively, to determine the prognostic factors in childhood meningitis in a developing country. Patients were examined for the following variables: history of antibiotic use; period between onset of symptoms and hospital admission; age at presentation; sex; fever; convulsion; level of consciousness; malnutrition; anemia; leukocyte and thrombocyte counts; erythrocyte sedimentation rate; serum C-reactive protein (CRP) level; and cerebrospinal fluid (CSF) including white blood cell count; glucose, protein, and CRP concentrations; antibiotic treatment; neurological sequelae; and fatality rate during the hospital stay. Most of these parameters were re-evaluated in all patients 36-48 h after admission. Patients were divided into 3 groups: surviving without sequelae, surviving with sequelae, and not surviving (deceased). A total of 48 children, 19 girls (39.5%) and 29 boys (60.5%), aged 2 months to 13 years, were included in the study. Of the 48 patients, 29 (60.5 %) survived without sequelae, 13 (27%) survived with sequelae and 6 (12.5%) died. In a comparison among groups, we found that absence of anemia, low (< 1,000) CSF white blood cell (WBC) count, and high CRP level at admission were the indicative of poor prognosis. Thirty-six to 48 h after admission, the presence of fever, depressed level of consciousness, high (> 1,000) CSF WBC count, and low CRP level were also poor prognostic factors. In addition, we observed that mortality rate was lower in the penicillin G + chloramphenicol group than in the ampicillin-sulbactam + cefotaxime group (P < 0.05). The mean period between onset of symptoms and hospital admission was longer in the surviving with sequelae and in the not surviving groups than in the surviving without sequelae group (P < 0.05). PMID- 12765224 TI - Enhancement of norepinephrine-induced transient contraction in aortic smooth muscle of diabetic mice. AB - Changes in norepinephrine-induced transient contractions in Ca2+-deficient solution were investigated in the aortic smooth muscles of diabetic ALS (alloxan induced diabetes susceptible) mice. The transient contractions in diabetic mice were significantly larger than those in normal mice. The longer incubation of the muscle preparations in Ca2+-deficient solution made the transient contractions smaller, probably due to the leakage and decrease in norepinephrine-releasable stored Ca2+. The rate of this reduction in contraction was slower in diabetic mice. These results suggest that the leakage of intracellular stored Ca2+ caused by extracellular Ca2+ deficiency is attenuated in diabetic mice, contributing to enhanced norepinephrine-induced transient contractions. PMID- 12765225 TI - Guidelines and shared care for asthma and COPD. AB - Shared-care constructions between general practitioners and pulmonary physicians are seemingly attractive for asthma and COPD patients. Thus they have to be implemented in further guidelines. However, anticipation that rapid changes will occur in treatment options towards optimal disease management justifies rapid adjustments in these strategies and requires investigations as to their ultimate benefit in disease outcome. PMID- 12765226 TI - A high prevalence of culture-positive extrapulmonary tuberculosis in a large Dutch teaching hospital. AB - BACKGROUND: In the Netherlands the incidence of tuberculosis (TB) has increased during the last decade. Growing immigration and international travel were important determining factors. To determine if this has resulted in altered clinical manifestations of the disease, we assessed the clinical spectrum of all TB cases diagnosed at our hospital in the period 1994 to 2000. METHODS: All culture-proven TB cases during the study period were retrospectively reviewed for clinical and demographic data. RESULTS: Sixty-five patients were identified. Solitary pulmonary TB was diagnosed in 33.9%, extrapulmonary TB in 51.8% and combined pulmonary and extrapulmonary TB in 14.3% of all cases. Patients were of foreign descent in 78.6% of all cases. Incidence peaked between 15 to 45 years. Decreased immunity was an important determining factor in the older patients. Presenting symptoms were mostly aspecific causing an important doctor's delay in establishing the diagnosis in 25%. Mortality was 3.6% and isoniazid resistance 3.6% CONCLUSIONS: Our data suggest an increase in the percentage of extrapulmonary TB concomitantly with an increasing percentage of patients of foreign descent. Because of aspecific presenting symptoms, TB was often diagnosed late. Treatment is mainly hindered by non-compliance and a high index of suspicion is necessary in making the diagnosis. PMID- 12765228 TI - Perianal ulcer and rash. Secondary syphilis. PMID- 12765227 TI - Referral and consultation in asthma and COPD: an exploration of pulmonologists' views. AB - BACKGROUND: The burden of asthma and chronic obstructive pulmonary disease (COPD) on national healthcare systems is expected to increase substantially in future years. Referral guidelines for general practitioners (GPs) and pulmonologists may lead to more efficient use of healthcare facilities. We explored the prevailing views of pulmonologists regarding referral and once-only consultation in asthma and COPD, and compared these views with recently published transmural referral guidelines for GPs and pulmonologists. METHODS: Cross-sectional multiple case study. Twenty-nine Dutch pulmonologists working at non-university hospitals or specialised chest clinics participated in group discussion sessions. RESULTS: The outcome of the discussions and recently published referral guidelines for GPs and pulmonologists showed considerable similarity, but also some marked discrepancies. During the discussions, the main points of disagreement among the pulmonologists were: 1) should GPs or pulmonologists add long-acting beta2 agonists to asthma treatment regimens; 2) should the current cut-off point 'predicted FEV1 <50%' for referral of COPD patients be increased to 60 or 70%; and 3) should an annual exacerbation rate of two episodes a year be used as an undifferentiated referral criterion for COPD patients? For asthma, proposed back referral (i.e. from pulmonologist to GP) criteria rested on: required dose of inhaled steroids, persistent need for long-acting beta2-agonists, duration of clinical stability and persistence of airway obstruction. Back-referral criteria for COPD rested on age, blood-gas abnormalities and ventilatory limitations. Primary care monitoring facilities and 'shared-care' constructions were considered to be facilitating conditions for back-referral. CONCLUSIONS: This explorative study provided insights into how pulmonologists visualise a rational referral policy for patients with asthma or COPD. These insights can be taken into consideration in future revisions of referral and back-referral guidelines for GPs and pulmonologists. PMID- 12765229 TI - Geriatric syndromes: medical misnomer or progress in geriatrics? AB - Both in geriatric and internal medicine journals, and in medical textbooks certain (aggregates of) symptoms are labelled as 'geriatric syndromes'. In frail elderly patients a large number of diseases present with well-known and highly prevalent atypical symptoms (e.g. immobility, instability, impaired cognition and incontinence), which are referred to as geriatric syndromes. While classically the term syndrome is used for grouping together multiple symptoms with a single pathogenetic pathway, geriatric syndrome primarily refers to one symptom or a complex of symptoms with high prevalence in geriatrics, resulting from multiple diseases and multiple risk factors. The geriatric workup should therefore consist of both a search for and treatment of the aetiologically related diseases and a risk factor assessment and reduction. Effectiveness and efficiency of this specific geriatric syndrome workup has been demonstrated predominantly for combinations of geriatric syndromes that often serve as targeting criteria for geriatric interventions, and for some specific geriatric syndromes. Therefore, we argue that the concept of geriatric syndromes is valuable as a theoretical frame, a directive for diagnostic analysis and as an educational tool in teaching geriatrics to medical students and trainees. Added to this, explaining the heterogeneous way 'syndrome' is used in current clinical practice, as opposed to 'disease', will also substantially improve clinical reasoning both in geriatrics and general internal medicine. PMID- 12765231 TI - Severe diffuse interstitial pneumonia due to Mycoplasma pneumoniae in a patient with respiratory insufficiency. AB - We report a 25-year-old man presenting with high fever, dyspnoea and somnolence. The presence of severe diffuse interstitial pneumonia with extrapulmonary symptoms, such as myositis and subclinical haemolysis, strongly suggested an infection by Mycoplasma pneumoniae. This diagnosis was supported by high titres of cold agglutinins and a positive Coombs test, and directly confirmed by specific IgM serological tests. After initiation of the appropriate antimicrobial treatment mechanical ventilation could be avoided and the patient showed a slow but complete clinical recovery. This diagnosis should be considered in any febrile patient with hypoxaemia and diffuse interstitial pneumonia, and rapid initiation of appropriate antibiotic treatment seems to be crucial for a favourable outcome. PMID- 12765230 TI - Fatal combination of moclobemide overdose and whisky. AB - The antidepressant moclobemide (Aurorix) is a reversible inhibitor of monoamine oxidase-A. Pure moclobemide overdose is considered to be relatively safe. Mixed drug overdoses including moclobemide are potentially lethal, especially when serotonergical drugs are involved. So far, only one fatality due to moclobemide mono-overdose has been reported. We report here on a fatality following the ingestion of a moclobemide overdose in combination with half a bottle of whisky. Although dietary restrictions during moclobemide therapy are not considered necessary, the combination of large quantities of moclobemide and tyramine containing products seems to be lethal, probably because monoamine oxidase-A selectivity is overwhelmed after massive overdoses. Since there is no specific antidote and treatment is only symptomatic, the severity of an overdose with moclobemide must not be underestimated. PMID- 12765232 TI - Coexistence of cystic intra-abdominal lymphangiomas and diffuse venous haemangiomas in adult life. AB - Diffuse haemangioma and intra-abdominal lymphangioma are rare in adults. In this case report, we present a 33-year-old female with coexisting multiple cutaneous and visceral cavernous haemangiomas and two huge intra-abdominal lymphangiomas of 25 and 35 cm in diameter. The organs involved were the liver, pericardium, renal hilus and bladder. She died due to disseminated intravascular coagulation and multiorgan failure, which resembled Kasabach-Merritt syndrome. The coexistence of generalised haemangiomas and intra-abdominal lymphangiomas and the lack of complaints until the age of 33 years makes her an unusual case in the literature. We also emphasise the other clinical conditions that should be considered in the differential diagnosis. PMID- 12765233 TI - Properties of phenyl valerate esterase activities from chicken serum are comparable with soluble esterases of peripheral nerves in relation with organophosphorus compounds inhibition. AB - Chicken serum, the usual in vivo animal for testing organophosphorus delayed neuropathy, has long been reported not to contain a homologous activity of the neuronal neuropathy target esterase (NTE) activity when it is assayed according to standard methods as the phenyl valerate esterase (PVase) activity, which is resistant to paraoxon and sensitive to mipafox. However, a PVase activity (1000 1500 nmol/min/ml) can be measured in serum that is extremely sensitive to both paraoxon, a non-neuropathic organophosphorus compound and mipafox, a model neuropathy inducer. The inhibition was time progressive in both cases, suggesting a covalent phosphorilating reaction. The fixed time inhibition curves suggest at least two sensitive components. The IC50 for 30 min, at 37 degrees C are 6 and 51 nM for paraoxon and 4 and 110 nM for mipafox, for every sensitive component. When paraoxon was removed from a serum sample pretreated with the inhibitor, the paraoxon sensitive PVase activity was recovered, in spite of showing a time progressive inhibition suggesting that hydrolytic dephosphorylating reaction recovered at a significant rate. The reactivation of the phosphorylated enzyme could explain that the time progressive inhibitions curves for long time with paraoxon tend to reach a plateau depending on the inhibition concentration. However, with mipafox, the curve approached the same maximal inhibitions at all concentrations as expected for a permanent covalent irreversible phosphorylation, which is coherent with the observations that the activity remained inhibited after removing the inhibitor. Data of serum esterases described in this paper showed similar properties to those previously reported for peripheral nerve soluble phenylvalerate esterase: (1) extremely high sensitivity to paraoxon and mipafox; (2) time progressive kinetic with two sensitive components; (3) recovery of activity after removal of paraoxon; and (4) permanent inhibition with mipafox. These properties of serum esterases are very similar to those of soluble fraction of peripheral nerves. So, serum PVases could be considered as appropriate biomarkers, as a mirror for the neural soluble paraoxon and mipafox sensitive soluble esterases that could be used for biomonitoring purpose. PMID- 12765234 TI - Property of metallothionein as a Zn pool differs depending on the induced condition of metallothionein. AB - Metallothionein (MT) is a low-molecular-weight, cysteine-rich, heavy metal binding protein with several putative physiological functions as a radical scavenger and a regulator of metabolism of Zn. Although the induction of hepatic MT by a wide range of stressors is a well-known phenomenon, little is known about the role of MT in stressful situations. Since MT regulates Zn metabolism, we investigated the differences between affinities of MT for Zn in various stressful conditions in this study. Divalent cadmium ions are capable of displacement of Zn from MT in vitro. Therefore, we assayed the binding of Zn to MT induced by various stimuli using in vivo administration of Cd. MT was induced by paraquat (PQ), a reactive oxygen generator, fasting stress and restraint stress. Apo-MT induced by them bound to Zn in vivo. Zn, which bound to MT induced by PQ, was displaced by the administration of Cd. However, Zn that had bound to MT induced by fasting stress and restraint stress was not displaced by Cd. Moreover, we assessed the in vivo affinity of Zn to MT induced by fasting stress under the condition of subchronic Cd exposure. Cd was administered to mice by subcutaneously implanted Alzet osmotic minipumps, which released constant amounts of Cd over a 14-day period. After 4 days, mice were fasted for 24 h and hepatic MT was examined. Interestingly, it was found that Zn had displaced Cd bound to MT. These findings indicate that the affinity of MT for Zn differs depending on the stimulus by which MT was induced. This is the first report on differences in the properties of MT depending on the stimulus used to induce MT. PMID- 12765235 TI - Lack of effects of sodium 2-mercaptoethane sulfonate (mesna) on Ochratoxin A induced renal tumorigenicity following life-time oral administration of Ochratoxin A in DA and Lewis rats. AB - Sodium 2-mercaptoethane sulfonate (Mesna) reacts with urotoxic metabolites of oxazaphosphorine drugs (e.g. cyclophosphamide or ifosfamide) and has been used clinically to protect against damage induced by these aggressive anti-neoplastic drugs in the kidney and lower urinary and genital tracts. Ochratoxin A (OTA) is a potent nephrotoxin in several species. In order to elucidate whether mesna has curative or preventive effects on OTA-induced renal damage or renal tumor development, we administered OTA and/or mesna to both DA and Lewis rats for their life-time and examined kidney, urethra and urinary bladder histologically. OTA induced sex- and strain-specific renal tumors. However, there was no evidence of any effect of mesna on the incidence and distribution of any type of tumor or non neoplastic finding in the kidney in either strain or treated group. In this study, we have confirmed that mesna treatment did not show any curative or preventive effects on either OTA-induced kidney damage or renal tumor development in two different strains that have distinct metabolic characteristics. PMID- 12765236 TI - The DNA methylation inhibitor 5-azacytidine modulates 6-thioguanine toxicity in mammalian cells. AB - In order to assess the effects of combining two antimetabolites used separately to treat human leukemias, we carried out an experimental study by exposing V79 Chinese hamster cells, a 6-thioguanine (6-tG)-sensitive cell line, to sequential and concurrent treatments with 5-azacytidine (5-azaC) and 6-tG. In this paper, we demonstrate that there is a clear dependency for the way in which this combination was tested. Pre-treatment with 5-azaC made V79 cells more resistant to 6-tG by a substantial reduction in 6-tG incorporation into DNA; this effect could still be detected for several cell divisions after the removal of 5-azaC, and was achieved neither by reduced cell growth nor by the induction of hypoxanthine-guanine-phosphoribosyltransferase (HGPRT-) mutants. The reverse order of treatment produced a higher toxic effect than exposure to each prodrug alone. PMID- 12765237 TI - Germ cell mutagenicity of gamma-ethyl-gamma-phenyl-butyrolactone (EPBL) detected in the CF1 mouse-dominant lethal study. AB - Female and male CFI mice weighing 25-30 g were given 0, 50, 100 or 200 mg/kg of gamma-ethyl-gamma-phenyl-butyrolactone (EPBL) for 5 days intraperitoneally. In the male-dominant lethal phase, males treated with EPBL were mated with untreated females following a 7-day mating schedule with three consecutive mating events. In the female-dominant lethal phase, females treated with EPBL were caged with untreated males. The above dosages and schedule treatments were used. The incidence of pregnancy of females mated on days 1-7 and 8-14 after males were given 200 mg/kg of EPBL and of females given 200 mg/kg when mated to untreated males was decreased. Upon examining surgically exposed uteri and ovaries of pregnant females during the first phase, on gestation days 13-15, an increased incidence of pre-implantation losses with 200 mg/kg of EPBL and an increased incidence of post-implantation losses with 100 and 200 mg/kg was observed. In addition, an increased frequency of pre- and post-implantation losses was seen in females treated with 200 mg/kg. These results support the conclusion that EPBL is a germ cell mutagen and its effects are more pronounced during the post-meiotic stage. PMID- 12765238 TI - The teratogenic potential of the herbicide glyphosate-Roundup in Wistar rats. AB - The aim of this study was to assess the teratogenicity of the herbicide glyphosate-Roundup (as commercialized in Brazil) to Wistar rats. Dams were treated orally with water or 500, 750 or 1000 mg/kg glyphosate from day 6 to 15 of pregnancy. Cesarean sections were performed on day 21 of pregnancy, and number of corpora lutea, implantation sites, living and dead fetuses, and resorptions were recorded. Weight and gender of the fetuses were determined, and fetuses were examined for external malformations and skeletal alterations. The organs of the dams were removed and weighed. Results showed a 50%, mortality rate for dams treated with 1000 mg/kg glyphosate. Skeletal alterations were observed in 15.4, 33.1, 42.0 and 57.3% of fetuses from the control, 500, 750 and 1000 mg/kg glyphosate groups, respectively. We may conclude that glyphosate-Roundup is toxic to the dams and induces developmental retardation of the fetal skeleton. PMID- 12765239 TI - Binding of polychlorinated biphenyls/metabolites to hemoglobin. AB - Polychlorinated biphenyls (PCBs) may form reactive electrophiles that can bind covalently to hemoglobin in vivo. Female C57/BL6 mice, pretreated with phenobarbital and P-naphthoflavone, were injected with either radiolabeled 4 chlorobiphenyl, 3,3',4.4'-tetrachlorobiphenyl, benzo[a]pyrene (B[a]P) or the corresponding non-labeled 4-chlorobiphenyl, 3.3',4.4'-tetrachlorobiphenyl, B[a]P or vehicle. Blood was collected at different time points. At 24 h a greater presence of 4-chlorobiphenyl and 3,3',4.4'-tetrachlorobiphenyl in the plasma, compared with the erythrocytes, was observed. For all three treatment groups the radioactivity in the hemolysate was found to be greater than the vehicle-treated group, except at 2 h after dosing, suggesting an association with hemoglobin. Covalent binding with globin was 10-fold greater for 4-chlorobiphenyl-treated animals compared with 3,3',4,4'-tetrachlorobiphenyl-treated animals. Liquid chromatography-mass spectral analysis of globin was used to identify adducts. Our preliminary data show an increase in mass corresponding to adducts of oxidized metabolites of PCBs. Detection of adducts of PCBs with hemoglobin could provide a valuable tool to evaluate acute exposure of a population. PMID- 12765240 TI - The multidrug resistance-associated protein 1 transports methoxychlor and protects the seminiferous epithelium from injury. AB - We examined the ability of the multidrug resistance-associated protein 1 (MRP1/ABCC1) to transport pesticides, as this transporter mediates the cellular efflux of a variety of xenobiotics, typically as glucuronide, sulfate, or glutathione conjugates. NIH3T3 cells stably expressing MRP1 were 3.37-fold more resistant to the toxicity of fenitrothion, 3.12-fold more resistant to chlorpropham, and 2.5-fold more resistant to methoxychlor, a pesticide with estrogenic and anti-androgenic metabolites. The cells expressing MRP1 also eliminated methoxychlor two times more rapidly than their mock-transfected counterparts. We then examined whether mrp1 expression could alter the toxicity of methoxychlor in vivo using male FVB/mrp1 knockout mice (FVB/mrp1-/-). Both control and knockout mice were fed 25 mg/kg methoxychlor in honey for 39 days, and its effects on testicular morphology were examined. Methoxychlor treatment did not significantly affect testicular morphology in the FVB mice, but markedly reduced the number of developing spermatocytes in the FVB/mrp1-/- mice. These results suggest that MRPI may play a role in protecting the seminiferous tubules from methoxychlor-induced damage. PMID- 12765241 TI - Expression of matrix metalloproteinase, tissue inhibitors of metalloproteinase and adhesion molecules in silicotic mice with lung tumor metastasis. AB - The number of metastatic foci in silicotic mice is approximately 1.5-fold that in normal mice and in mice treated with TiO2 as inert particles. Expression of matrix metalloproteinases (MMPs) tissue inhibitors of metalloproteinases (TIMPs) and selectins was investigated in silicotic mice with lung tumor metastasis. Expression of MMP-9 and P-selectin mRNA, but not MMP-2 and E-selectin, increased significantly, showing decreases of the ratio of expression in TIMPs/MMP-9 in tumor-bearing silicotic mice compared with the tumor-bearing normal mice and mice treated with TiO2. Pretreatment with anti-P-selectin antibody inhibited number of metastatic foci significantly in silicotic mice, while pretreatment of animals with anti MMP-9 antibody showed slight decrease of metastatic foci. This evidence indicated that up-regulation of P-selectin expression contributed to enhanced rate of tumor metastasis in lung with silicosis. PMID- 12765242 TI - ZnCl2 induces Syrian hamster embryo (SHE) cell transformation. AB - In order to test the hypothesis of a relationship between apoptosis and neoplastic transformation, we studied the transforming potency of zinc, known for its antiapoptotic effects. In this study, zinc chloride (100 microM) was shown to induce morphological transformation (MT) in Syrian hamster embryo (SHE) cells. It was also tested in combination with benzo(a)pyrene (BaP), a positive control for carcinogenicity, or fomesafen, a carcinogenic pesticide with hepatic peroxisomal proliferation properties. A co-exposure of the two carcinogens with 100 microM zinc increased cell transformation in SHE cells. These results were in agreement with the theory of a relationship between the inhibition of apoptosis and induction of cell transformation. The cloning efficiency (CE) of SHE cells seeded at clonal density was raised by zinc, fomesafen and furthermore by the mixture of the two chemicals, which could be explained by the antiapoptotic action of zinc and fomesafen on SHE cells. No change in myc and bax expressions was observed in zinc-treated SHE cells. PMID- 12765243 TI - Activation of estrogen receptor alpha and ERbeta by 4-methylbenzylidene-camphor in human and rat cells: comparison with phyto- and xenoestrogens. AB - 4-Methylbenzylidene-camphor (4-MBC) is an organic sunscreen that protects against UV radiation and may therefore help in the prevention of skin cancer. Recent results on the estrogenicity of 4-MBC have raised concerns about a potential of 4 MBC to act as an endocrine disruptor. Here, we investigated the direct interaction of 4-MBC with estrogen receptor (ER) alpha and ERbeta in a series of studies including receptor binding, ER transactivation and functional tests in human and rat cells. 4-MBC induced alkaline phosphatase activity, a surrogate marker for estrogenic activity, in human endometrial Ishikawa cells. Interestingly, 4-MBC induced weakly ERalpha and with a higher potency ERbeta mediated transactivation in Ishikawa cells at doses more than 1 microM, but showed no distinct binding affinity to ERalpha or ERbeta. In addition, 4-MBC was an effective antagonist for ERalpha and ERbeta. In an attempt to put 4-MBC's estrogenic activity into perspective we compared binding affinity and potency to activate ER with phyto- and xenoestrogens. 4-MBC showed lower estrogenic potency than genistein, coumestrol, resveratrol, bisphenol A and also camphor. Analysis of a potential metabolic activation of 4-MBC that could account for 4-MBC's more distinct estrogenic effects observed in vivo revealed that no estrogenic metabolites of 4-MBC are formed in primary rat or human hepatocytes. In conclusion, we were able to show that 4-MBC is able to induce ERalpha and ERbeta activity. However, for a hazard assessment of 4-MBC's estrogenic effects, the very high doses of 4-MBC required to elicit the reported effects, its anti estrogenic properties as well as its low estrogenic potency compared to phytoestrogens and camphor has to be taken into account. PMID- 12765244 TI - Transplacental pharmacokinetics of the new fluoroquinolone DW-116 in pregnant rats. AB - DW-116 is a newly developed fluoroquinolone antibacterial with a broad spectrum against both gram-positive and gram-negative bacteria. Recently, we have reported that DW-116 induces a significant developmental toxicity in rat. The present study was undertaken to characterize the placental transfer and pharmacokinetics of DW-116 in Sprague-Dawley rats. Pregnant females were given a single oral dose of 500-mg [14C]DW-116/kg on gestational day 18. Maternal and fetal tissues were collected at 0.17, 0.5, 1, 2, 4, 8 and 24 h after dosing. The [14C]DW-116-derived radioactivity was rapidly distributed to the fetus and slowly eliminated from the tissue. The radioactivity in both maternal plasma and fetal tissue reached its peak within 1 h and maintained the level of radioactivity up to 16-28% of the peak level until 24 h after dosing. Radioactivity in whole fetus was higher than those in the maternal plasma and placenta. The T(1/2)abs, T(1/2)beta, AUC, Tmax and Cmax in the maternal plasma were approximately 6 min, 13.3 h, 1620 microg h/ml, 0.5 h and 136 microg/ml, respectively. Those in the placenta were approximately 20 min, 12.3 h, 2150 microg h/ml, 1.0 h and 172 microg/ml, respectively. Those in the whole fetus were 13 min, 12.8 h, 2549 microg h/ml, 1 h and 191 microg/ml, respectively. In the amniotic fluid of maternal uterus, the T(1/2)abs, T(1/2)beta, AUC, Tmax and Cmax were approximately 1.3 h, 9.3 h, 2508 microg h/ml, 4.4 h, and 135 microg/ ml, respectively. While DW-116 disappeared biphasically from maternal plasma, whole fetus and placenta, it was eliminated monophasically from amniotic fluid. These results indicate that (1) total radioactivity appeared rapidly in maternal plasma and fetuses; (2) the elimination of total radioactivity is slow; and (3) DW-116 or relevant metabolites could cross the blood-placenta barrier in pregnant rats. PMID- 12765245 TI - Mutation analysis of vinyl carbamate or urethane induced lung tumors in rasH2 transgenic mice. AB - Previous studies showed that significant differences in mutation frequency of the human c-Ha-ras transgene between vinyl carbamate (VC)- and ethyl carbamate (urethane)-induced lung tumors were observed in rasH2 mice. It remains unclear why the point mutation frequency is extremely low in VC-induced lung tumors, although this compound is much more carcinogenic than urethane. In this study, we examined the somatic point mutations of the transgene at the RNA level in VC- and urethane-induced lung tumors of rasH2 mice. We did not find any mutation at codon 12 of the transgene in any of these lung tumors, but codon 61 showed frequent mutations in not only urethane-induced lung tumors (15 out of 16) but also VC induced lung tumors (11 out of 11) in rasH2 mice. These results suggested that point mutations at codon 61 of the transgene play an important role in the carcinogenesis of VC- and urethane- induced lung tumors in rasH2 mice. PMID- 12765246 TI - Comparison of the reporter gene assay for ER-alpha antagonists with the immature rat uterotrophic assay of 10 chemicals. AB - We performed a reporter gene assay for estrogen receptor (ER)-alpha agonists and antagonists of 10 chemicals that showed both estrogen agonistic and reduced the estrogenic effect of ethinyl estradiol in a rat uterotrophic assay. The chemicals tested by the immature uterotrophic assay were p-(tert-pentyl)phenol, 4,4' thiobis-phenol, 4,4'-(hexafluoroisopropylidene)diphenol, 2,2-bis(4-hydroxyphenyl) 4-methyl-n-pentane, 4,4'-(octahydro-4,7-methano-5H-inden-5-ylidene)bisphenol, 4 (phenylmethyl)phenol, 4,4'-dihydroxybenzophenone, 2,2',4,4' tetrahydroxybenzophenone, 4-hydroxybenzophenone and 2,4,4' trihydroxybenzophenone. Although all chemicals examined in this study were positive in the reporter gene assay for ER-alpha agonists, 4,4'-(octahydro-4,7 methano-5H-inden-5-ylidene)bisphenol was only positive in the reporter gene assay for ER-alpha antagonists. These findings demonstrate that results of the reporter gene assay for ER-alpha agonists correlated well with those of the uterotrophic assay, but antagonistic change of 9 of 10 chemicals in the uterotrophic assay was not detected by the reporter gene assay for ER-alpha antagonists. PMID- 12765247 TI - The effect of m-xylene on cytotoxicity and cellular antioxidant status in rat dermal equivalents. AB - Exposure of the skin to volatile organic chemicals (VOCs) can lead to irritation, inflammation and cytotoxicity. Since VOCs are used in industrial, commercial and military applications, concern is mounting with respect to VOC safe exposure limits. Although traditional toxicological assessment of VOCs has utilized animal models, the use of alternative in vitro models is becoming more widespread. We have previously developed a sealed exposure system that prevents chemical loss through evaporation and enables calculation of target cell chemical dose. The present study utilized this in vitro exposure method to assess m-xylene-induced cytotoxicity and antioxidant status in dermal equivalents (dermal fibroblasts in a collagen matrix). At the end of a 1- or 4-h exposure, cytotoxicity was measured using the MTT assay and the EC50 values determined were 1481 +/- 88 and 930 +/- 33, respectively. Decreases in cellular thiols and catalase activity were observed, which occurred in a time and dose-dependent manner. Treatment of dermal equivalents with the antioxidants N-acetylcysteine (NAC) and catalase provided some protection against m-xylene-induced cytotoxicity. When compared to m-xylene exposures, treatment with either 1.0 or 5.0 mM NAC led to increases in the EC50 values at 1 and 4 h. Increases in these EC50 values ranged from 1.22- to 1.32 fold at 1 h and 1.27- to 1.54-fold at 4 h. Although treatment with catalase (1000 U/ml) led to a 1.35-fold increase in cell viability at 1 h, no significant differences were observed at either 1 or 4 h when compared to dermal equivalents exposed to m-xylene alone. These results suggest that exposure to m-xylene leads to a time- and dose-dependent decrease in cellular antioxidants and that cellular thiols may provide protection against the cytotoxic properties of m-xylene. PMID- 12765249 TI - Energy metabolism and the evolution of reproductive suppression in the human female. AB - Reproduction places severe demands on the energy metabolism in human females. When physical work entails higher energy expenditure, not enough energy will be left for the support of the reproductive processes and temporal suppression of the reproductive function is expected. While energy needed for reproduction may be obtained by increases in energy intake, utilization of fat reserves, or reallocation of energy from basal metabolism, several environmental or physiological constraints render such solutions unlikely. For human ancestors increases in energy intake were limited by availability of food, by labor of food preparation and by metabolic ceilings to energy assimilation. Energy stored as fat may support only a fraction of the requirements for reproduction (especially lactation). Effects of intense physical activity on basal metabolism may also interfere with fat accumulation during pregnancy. Finally, the female physiology may experience demands on increasing the basal metabolism as a consequence of physical activity and, at the same time, on decreasing the basal metabolism, when energy to support the ongoing pregnancy or lactation is inadequate. The resulting metabolic dilemmas could constitute a plausible cause for the occurrence of reproductive suppression in response to physical activity. It is, therefore, likely that allocating enough energy to the reproductive processes during periods when energy expenditure rises may be difficult due to physiological and bioenergetic constraints. Females attempting pregnancy in such conditions may compromise their lifetime reproductive output. A reproductive suppression occurring in low energy availability situations may thus represent an adaptive rather then a pathological response. PMID- 12765248 TI - Warifteine and milonine, alkaloids isolated from Cissampelos sympodialis Eichl: cytotoxicity on rat hepatocyte culture and in V79 cells. AB - Two alkaloids were isolated from the leaves of Cissampelos sympodialis; a bisbenzylisoquinoline compound named warifteine and a novel 8,14 dihydromorphinandienone alkaloid named milonine. The cytotoxic effects of these alkaloids were assayed in cultured hepatocytes and V79 fibroblasts. Three independent endpoint assays for cytotoxicity in vitro were used: the nucleic acid content (NAC), tetrazolium reduction (MTT) and neutral red uptake (NRU). Milonine was less toxic than warifteine in both cell cultures. The IC50 values determined in the three different viability assays were around 100 and 400 microM after milonine treatment of V79 cells or hepatocytes. IC50 values ranging from 10 to 35 microM were obtained for warifteine in the viability tests evaluated in V79 cells and hepatocytes. Due to the similar cytotoxic effects detected on V79 cells and hepatocytes, probably warifteine and milonine induced toxic effects independent to the cytochrome P450. This hypothesis was corroborated by the results where Cimetidine (1.0 mM), a traditional cytochrome P450 inhibitor, did not protect the cells from the toxic action of warifteine or milonine. In conclusion, these alkaloids merit further investigations as potential novel pharmacological agents although milonine was less toxic than warifteine in the cells models investigated. PMID- 12765250 TI - A mathematical model for the spontaneous contractions of the isolated uterine smooth muscle from patients receiving progestin treatment. AB - The in vitro spontaneous contractions of human myometrium samples can be described using a phenomenological model involving different cell states and adjustable parameters. In patients not receiving hormone treatment, the dynamic behavior could be described using a three-state model similar to the one we have already used to explain the oscillations of intrauterine pressure during parturition. However, the shape of the spontaneous contractions of myometrium from patients on progestin treatment was different, due to a two-step relaxation regime including a latched phase which cannot be simulated using the previous model without introducing an ad hoc mechanism to account for the extra energy involved in this sustained contraction. One way to do this is to introduce an anomalous rate of ATP consumption, the biochemical reasons for which have not yet been elucidated and which cannot be mathematically simulated using our experimental data. An alternative explanation is the reduced cycling rate of actin-myosin cross-bridges known to occur during the latch-phase. Our experimental findings suggest a third possibility, namely a sol-gel transition with a specific relaxation time constant, which would maintain a significant part of the cell population in the contracted-state until the intracellular-medium returns to its normal fluid behavior. PMID- 12765251 TI - An information-theoretical measure of taxonomic diversity. AB - Traditional diversity indices are computed from the abundances of species present and are insensitive to taxonomic differences between species. However, a community in which most species belong to the same genus is intuitively less diverse than another community with a similar number of species distributed more evenly between genera. In this paper, we propose an information-theoretical measure of taxonomic diversity that reflects both the abundances and taxonomic distinctness of the species. Unlike previous measures of taxonomic diversity, such as Rao's quadratic entropy, in this new measure the analyzed taxonomic properties are associated with the single species instead of species pairs. PMID- 12765252 TI - Theoretical evidence that more microtubules reach the cortex at the pole than at the equator during anaphase in sea urchin eggs. AB - Astral microtubules are rapidly elongated during anaphase and telophase in sea urchin eggs. The number of microtubules extending to the cell surface was calculated with a computer. For the calculations, microtubules were assumed to radiate from the astral center uniformly over angles. Although microtubules from two asters freely overlapped around the equator, the number per the unit area, i.e. the surface density, was larger in the polar region than in the equatorial region. The ratio of the theoretically calculated numbers in the two regions was close to the ratio obtained from the ultrastructural observations by Asnes and Schroeder in 1979. When counted in the longitudinal section including two astral centers, the microtubule number was a little larger in the equatorial region than in the polar region. However, the numbers do not represent the surface density because the two-dimensional section contains only a small portion of all the microtubules spreading in the three-dimensional space. The fluorescence image for tubulin, in most cases, provides the microtubule distribution in the longitudinal section. Therefore, from such an image, we cannot judge whether the surface density of astral microtubules is larger at the pole or at the equator. PMID- 12765253 TI - Hydrological adjustment and flooding control of wetlands in the Liaohe Delta. AB - The function of estuary wetland on hydrological adjustment and flooding control is studied in this paper. It is estimated that the evapotranspiration in the reed field during growth season (June to October) is 722.9 mm, which is 37.5% higher than large water body (E601: 525.9 mm). The water replacement rate in the reed field can reach 95% only when the rains continuously for 11 days and the precipitation reached 912 mm. For the water balance in the paddy field, the total water requirement ranges between 1920 and 1860 mm, among which, 31% is from precipitation, and the left is provided by reservoirs. The water usage efficiency is 0.35 at present productivity. Based on the landscape characteristics and functionalities on flooding control, 5 functional zones are designed for the Liaohe Delta; key protected area; underground storage area; flooding discharge area; flood diversion area in emergency; and flood control drainage area. PMID- 12765254 TI - Natural and artificial landscape change in a Dutch estuary: partially monitored with low budget method (a study in the fourth dimension). AB - This study includes some aspects of the shift in the Dutch attitude in relation to water during the past millennia from defense to attack to keeping the balance ("co-evolution"). It has a special focus on the freshwater tidal part, which embraces the largest seaport of the world: Rotterdam, as well as the largest national park of The Netherlands. It reports especially about a young mans endeavor in half a century real time monitoring of some land(scape) units with simple means. PMID- 12765255 TI - Roles of urban vegetation on balance of carbon and oxygen in Guangzhou, China. AB - The plant biomass and net primary production (NPP) of urban vegetation in Guangzhou were estimated by dimension analysis, tree truck volume, and harvest methods as well as relationship between biomass and NPP and so on. The biomass and NPP were respectively 2875150t and 1058122 t/a. They were respectively 392495t and 64948 t/a in the built-up area and 2482655t and 993147 t/a in the unbuilt-up area. It would make plant biomass, especially NPP decline obviously, if the unbuilt-up area were changed to the built-up area. The carbon content of plant was 1328649 for the total and 13.78 t/hm2 for the mean, and amounts of carbon fixed and oxygen made by urban vegetation were respectively 4.80 t/(hm2 x a) and 12.79 t/(hm2 x a) for the mean and 462624 t/a and 1232430 t/a for the total, which were equal to 1.45 times and 1.04 times of those by human breathing. However, they were only equal to 7.61% and 4.97% of amount of carbon released and oxygen consumption in urban Guangzhou. The biomass and NPP of urban vegetation in Guangzhou only corresponded to 7.8% and 47.3% of those of southern subtropical evergreen broad-leaf forest in Dinghu Mountain. Therefore, the roles of Guangzhou urban vegetation in balance of carbon and oxygen would be increased greatly if it could be conserved and improved in some way. PMID- 12765256 TI - River system in Japan from a landscape ecological aspect. AB - The objective of this study was to elucidate characteristics associated with rivers by classification of major rivers in Japan into several types based on riparian conditions. As the results of principal component analysis (PCA) with use of parameters reflecting form and artificial alterations of respective rivers, four major components such as "comprehensive riparian size", "intactness of water front", "continuity of streams" and "simplicity of landform in river basin" were extracted. Subsequently, cluster analysis was performed based on principal component scores, leading to successful classification of major rivers into 6 types. These findings disclosed that (1) the extracted principal components provide effective viewpoint for classification of rivers; (2) distribution of respective classes indicates area properties; and (3) the employed quantitative procedures were found effective for classification of major rivers. PMID- 12765258 TI - Ecological networks and greenways in Europe: reasoning and concepts. AB - The paper gives an overview of approaches towards ecological networks throughout Europe. It does not intend to present a complete picture, but to highlight common developments within countries and regions and show common principles and differences between countries and regions that have to be taken into account when developing a joint European initiative. Countries or regions that have not been included can be active in the same way, but information was not accessible to the authors for different reasons. This overview shows the comparable trends in decline of landscapes and the diversity in approaches to biodiversity conservation and nature conservation planning. Understanding the difference and common issues are of utmost importance to generalise common principles and understand the way neighbours and other European partners approach problems. PMID- 12765257 TI - Comparing the soil quality changes of different land uses determined by two quantitative methods. AB - Soil quality is one of the most important environmental factors in sustaining the global biosphere and developing sustainable agricultural practices. A study was initiated in Wolong Nature Reserve, Sichuan Province, China to elucidate the soil quality changes of natural secondary succession, forest planting and agricultural practices after deforestation in the humid mountainous region. The soil qualities of six land use types (natural forestland, grassland, shrub land, secondary forestland, cultivated land and reforested land) were compared using two quantitative methods: the integrated soil quality index (QI) and soil deterioration index (DI). The QI values of natural forestland, grassland, shrub land, secondary forestland, cultivated land, reforested land were 0.8039, 0.3277, 0.9127, 0.6881, 0.0285 and 0.3183, respectively. The DI values were 0%, -14%, 12%, 1%, -26% and -18% respectively. Both indexes suggested that shrub land can restore soil properties. To compare the two methods more directly, a deduced index QI' based on QI value was developed. The results showed that DI and QI' had a very high linear correlation coefficient (r = 0.9775) despite the values were different. Both methods were efficient in evaluating the soil quality levels and DI was a more simple way in soil quality assessment, while QI could show more ecological meanings. PMID- 12765259 TI - Detecting forest landscape boundary between mountain birch and evergreen coniferous forest in the northern slope of Changbai Mountain. AB - Boundaries between different forest types in Changbai Mountain Eastern China are results from complex interactions between forest ecosystems, topography, and geomorphology. Detecting and quantifying the transitional zones are highly important since high environmental heterogeneity and biodiversity are often found within these zones. In this study, we used GIS and multivariate statistics techniques (PCA and MSWA) to analyze data from Landsat TM satellite imageries and quantitatively determined the positions and widths of the landscape boundary between mountain birch and evergreen coniferous forests in the northern slope of Changbai Mountain. The results showed that the widths of the landscape boundary ranges from 30-50 m while using the MSWA or/and PC method. Such detected widths are consistent with field transect data that suggests a 50 m transitional zone width. The results further suggest that TM data can be used in combination with GIS and statistical techniques in determining forest landscape boundaries; MSWA is more reliable than PCA, while PCA can also be used to determine the landscape boundary when transects are properly located. PMID- 12765260 TI - Multifunctional landscapes--perspectives for the future. AB - New methods in landscape ecology to study the link between landscape heterogeneity and landscape functionality are needed. Heterogeneity is a basic characteristic of landscape, and landscape function is the capacity to change the structural heterogeneity of a landscape system. In most developed countries the industrialisation of agriculture has in general resulted in a change of agricultural landscapes from a small-grained heterogeneous pattern towards more monotonous and monofunctional landscapes. During the 1990's this trends seem to have changed due to a diversification of rural land use, and new trends in ubanisation. Whether these phases of landscape development should be expected in developing countries is a totally open question. Dealing with the study of multifunctionality of landscapes it is proposed to distinguish between ecological functionality of landscape ecosystems, functionality pertaining to land use and social functionality. Further, the relation between function, space and scale is important by the determination of spatial and time segregation as well as spatial and time integration of multifunctionality in landscapes. PMID- 12765261 TI - Study on the oasis corridor landscape in the arid regions based on RS and GIS methods--application of Jinta Oasis, China. AB - The study on the oasis corridor landsape is a new hotspot in the ecological environment research in the arid regions. In oasis, main corridor landscape types include river, ditch, shelterbelt and road. This paper introduces the basic ecological effects of the corridor landscape on the transporting mass and energy and obstructing desert landscape expansion and incursion. Using Geographic Information System (GIS), we have researched the corridor distribution and its spatial relationship with other landscape types in the Jinta Oasis. Based on the dynamically monitoring on the landscape pattern change of the Jinta Oasis during the latter 10 years by using Remote Sensing (RS) and GIS, the driving functions of the corridors on this change have been analyzed in detail. The analysis results showed that all kinds of corridors' characteristics can be quantified by the indexes such as length and width, ratio of perimeter and area, density and non-heterogeneity. The total corridor length of Jinta Oasis is 1838.5 km and its density is 2.1 km/km2. The corridor density of the irrigation land, forest and resident area is maximal, which shows that affection degree of the oasis corridors on them is the most. The improvement of the corridors quality is one of the important driving factors on the irrigation land and so on. The organic combination of the RS and GIS technologies and landscape research methods would be an effective means for the corridor landscape research on arid region oasis. PMID- 12765262 TI - Landscape pattern and its effect on ecosystem functions in Seoul Metropolitan area: urban ecology on distribution of the naturalized plant species. AB - During land transformation process in the human history, naturalized plants were introduced to several land use patterns by the different ways of plant itself. Including some naturalized plants that had been contribute to land restoration, many naturalized plants have been invaded to original habitat or landscape for native plants. Once the plants were colonized, they extend their area and population size. Urban developed areas often give an important role of source habitat for naturalized plants and expanding their population size. In recent, this situation is appearing as one of environmental problem about the urban landscape management controlling the naturalized plants that invaded in the developed area and conserving the native vegetation. This paper is focusing on relationships between distribution of habitat of naturalized plants and landscape patch in urban areas in Seoul. Gangdong-Gu, one of the administrative areas in Seoul was selected for this study. We examined the recent land use change using LANDSAT TM data and spreading of the representative naturalized plants (Robinia pseudocacia and Eupatorium rugosum) by Seoul Biotope Mapping Project and field survey in 1999. As a result, these two species were often occurred in the same habitat and distributed in forest edge disturbed by man. Their distribution patterns were related to landscape indices (patch size and shape) in the forest edge. PMID- 12765263 TI - The impact of urban planning on land use and land cover in Pudong of Shanghai, China. AB - Functional zones in cities constitute the most conspicuous components of newly developed urban area, and have been a hot spot for domestic and foreign investors in China, which not only show the expanse of urban space accompanied by the shifts both in landscape (from rural to urban) and land use (from less extensive to extensive), but also display the transformation of regional ecological functions. By using the theories and methods of landscape ecology, the structure of landscape and landscape ecological planning can be analyzed and evaluated for studying the urban functional zones' layout. In 1990, the Central Government of China declared to develop and open up Pudong New Area so as to promote economic development in Shanghai. Benefited from the advantages of Shanghai's location and economy, the government of Pudong New Area has successively built up 7 different functional zones over the past decade according to their functions and strategic goals. Based on the multi-spectral satellite imageries taken in 1990, 1997 and 2000, a landscape ecology analysis was carried out for Pudong New Area of Shanghai, supported by GIS technology. Green space (including croplands) and built-up area are the major factors considered in developing urban landscape. This paper was mainly concerned with the different spatial patterns and dynamic of green space, built-up areas and new buildings in different functional zones, influenced by different functional layouts and development strategies. The rapid urbanization in Pudong New Area resulted in a more homogeneous landscape. Agricultural landscape and suburban landscape were gradually replaced by urban landscape as the degree of urbanization increased. As consequence of urbanization in Pudong, not only built-up patches, but also newly-built patches and green patches merged into one large patch, which should be attributed to the construction policy of extensive green space as the urban development process in Pudong New Area. The shape of green area of 7 functional zones became more and more regular because of the horticultural needs in Shanghai urban planning. Some suggestions were finally made for the study of future urban planning and layout. PMID- 12765264 TI - Analysis on sensitivity and landscape ecological spatial structure of site resources. AB - This article establishes a set of indicators and standards for landscape ecological sensitivity analysis of site resources by using the theories and approaches of landscape ecology. It uses landscape diversity index (H), evenness (E), natural degree (N), contrast degree (C) to study spatial structure and landscape heterogeneity of site resources and thus provides a qualitative quantitative evaluation method for land planning and management of small, medium scale areas. The analysis of Yantian District, Shenzhen of China showed that Wutong Mountain belonged to high landscape ecological sensitivity area, Sanzhoutian Reservoir and Shangping Reservoir were medium landscape sensitivity and high ecological sensitivity area; Dameisha and Xiaomeisha belonged to medium sensitivity area caused by the decline of natural ecological areas. Shatoujiao, Yantian Pier belonged to low sensitivity area but urban landscape ecological development had reshaped and influenced their landscape ecological roles in a great extent. Suggestions on planning, protection goals and development intensity of each site or district were raised. PMID- 12765265 TI - Landscape dynamitic change in Mu Us Desert derived from Landsat TM data. AB - Landscape ecology emphasizes large areas and ecological effects of the spatial patterning of ecosystem. Recent developments in landscape ecology have emphasized the important relationship between spatial patterns and many ecological processes. Quantitative methods in landscape ecology link spatial patterns and ecological processes at broad spatial and temporal scales. In turn the increased attention on temporal change of ecosystem has highlighted the need for quantitative methods that can analyze patterns. This research applies quantitative methods--change detection to assess the ecosystem temporal change in the arid and semiarid area. Remote sensing offers the temporal change of ecosystem on landscape characteristics. PMID- 12765266 TI - Spatial analysis of plant detritus processing in a Mediterranean river type: the case of the River Tirso Basin, Sardinia, Italy. AB - The river continuum concept represents the most general framework addressing the spatial variation of both structure and function in river ecosystems. In the Mediterranean ecoregion, summer drought events and dams constitute the main sources of local disturbance to the structure and functioning of river ecosystems occurring in the river basin. In this study, we analysed patterns of spatial variation of detritus processing in a 7th order river of the Mediterranean ecoregion (River Tirso, Sardinia-Italy) and in three 4th order sub-basins which were exposed to different summer drought pressures. The study was carried out on Phragmites australis and Alnus glutinosa leaf detritus at 31 field sites in seasonal field experiment. Detritus processing rates were higher for Alnus glutinosa than for Phragmites australis plant detritus. Processing rates of Alnus glutinosa leaves varied among seasons and study sites from 0.006 d(-1) to 0.189 d(-1) and those of Phragmites australis leaves ranged from 0.0008 d(-1) to 0.102 d(-1), with the lowest values occurring at sites exposed to summer drought. Seasons and sites accounted for a significant proportion of such variability. Alder detritus decay rates generally decreased with increasing stream order, while reed detritus decay rates generally increased on the same spatial gradient. Summer drought events affected these spatial patterns of variation by influencing significantly the decay rates of both plant detritus. The comparisons among and within sub-basins showed strong negative influence of summer drought on detritus processing rates. Similarly, in the entire River Tirso basin decay rates were always lower at disturbed than at undisturbed sites for each stream order; decay rates of reed detritus remained lower at those sites even after the end of the disturbance events, while alder decay rates recovered rapidly from the summer drought perturbations. The different recovery of the processing rates of the two leaves could also explain the different patterns of spatial variation observed between the two leaves. PMID- 12765267 TI - Landscape structure and bird's diversity in the rural areas of Taiwan. AB - This study tries to discuss the relationship between landscape structure and organisms in the perspective of landscape architecture. The research hypotheses were then proposed as (1) there are relationships between landscape structure indexes and birds' diversity in the rural areas of Taiwan; (2) the relationships between landscape structure and birds' diversity will be different in different hierarchical levels. In order to increase the bird species, landscape planners could tries to increase the density of water bodies, but decrease the farms and human planted woods. Decrease the density of constructed and human planted grasslands. Increase the area of un-worked areas, natural grasslands, and the area of water bodies and circular the water bodies and natural forest. In order to increase birds' diversity, landscape planners could decrease the concentration of paved areas. Concentrate the human planted trees to increase the core areas of woodlands. Increase the area of natural grassland circular. In order to increase the total number of birds in the planning areas, landscape planners could scattered the paved areas and lengthen the constucted areas. Decreases the core region of the constructed areas. Increase the area of un-worked acres and water bodies. Decrease the disturbance of both the interior area of natural and human planted woodlands and try to increase the density of water bodies. The analysis results showed that the small grain size indexes are more suitable for the rural areas of Taiwan to capture the influential factors of bird communities. The high fragmentation of land usages in Taiwan lessens the influences of the regional landscape pattern. PMID- 12765268 TI - The effects of drainage basin geomorphometry on minimum low flow discharge: the study of small watershed in Kelang River Valley in Peninsular Malaysia. AB - This study investigate the relationships between geomorphometric properties and the minimum low flow discharge of undisturbed drainage basins in the Taman Bukit Cahaya Seri Alam Forest Reserve, Peninsular Malaysia. The drainage basins selected were third-order basins so as to facilitate a common base for sampling and performing an unbiased statistical analyses. Three levels of relationships were observed in the study. Significant relationships existed between the geomorphometric properties as shown by the correlation network analysis; secondly, individual geomorphometric properties were observed to influence minimum flow discharge; and finally, the multiple regression model set up showed that minimum flow discharge (Q min) was dependent of basin area (AU), stream length (LS), maximum relief (Hmax), average relief (HAV) and stream frequency (SF). These findings further enforced other studies of this nature that drainage basins were dynamic and functional entities whose operations were governed by complex interrelationships occurring within the basins. Changes to any of the geomorphometric properties would influence their role as basin regulators thus influencing a change in basin response. In the case of the basin's minimum low flow, a change in any of the properties considered in the regression model influenced the "time to peak" of flow. A shorter time period would mean higher discharge, which is generally considered the prerequisite to flooding. This research also conclude that the role of geomorphometric properties to control the water supply within the stream through out the year even though during the drought and less precipitations months. Drainage basins are sensitive entities and any deteriorations involve will generate reciprocals and response to the water supply as well as the habitat within the areas. PMID- 12765269 TI - The effects of land use and its patterns on soil properties in a small catchment of the Loess Plateau. AB - Due to relatively strong human activities in the hilly area of Loess Plateau, the natural vegetation has been destroyed, and landscape pattern based on agricultural land matrix was land use mosaic composing of shrub land, grassland, woodland and orchard. This pattern has an important effect on soil moisture and soil nutrients. The Danangou catchment, a typical small catchment, was selected to study the effects of land use and its patterns on soil moisture and nutrients in this paper. The results are as follows: The comparisons of soil moisture among seven land uses for wet year and dry year were performed; (1) the average of soil moisture content for whole catchment was 12.11% in wet year, while it was 9.37% in dry year; (2) soil moisture among seven land uses significantly different in dry year, but not in wet year; (3) from wet year to dry year, the profile type of soil moisture changed from decreasing type to fluctuation-type and from fluctuant type to increasing type; (4) the increasing trend in soil moisture from the top to foot of hillslope occurred in simple land use along slope, while complicated distribution of soil moisture was observed in multiple land uses along slope. The relationship between soil nutrients and land uses and landscape positions were analysed: (1) five nutrient contents of soil organic matter (SOM), total N (TN), available N (AN), total P (TP) and available P (AP) in hilly area were lower than that in other area. SOM content was less than 1%, TN content less than 0.07%, and TP content between 0.05% and 0.06%; (2) SOM and TN contents in woodland, shrub land and grassland were significantly higher than that in fallow land and cropland, and higher level in soil fertility was found in crop-fruit intercropping land among croplands; (3) soil nutrient distribution and responses to landscape positions were variable depending on slope and the location of land use types. PMID- 12765270 TI - Fragmented forest in tropical landscape--the case of the state of Selangor, Malaysia. AB - This paper presents the pattern and changes of fragmented forest in relation with changes of total forest cover in the state of Selangor in three decades. In this study, inventoried forest cover maps of Selangor in 1971/1972, 1981/1982 and 1991/1992 produced by the Forestry Department of Peninsular Malaysia were digitized to examine the changes in area and number of fragmented forest. Results showed that in 1971/1972, 16 fragmented forests were identified in Selangor. All fragmented forests were identified as dipterocarp forest. A decade later the number of fragmented forests increased by approximately 44% (23). Of the 23 fragmented forests, two were peat swamp forests whereas the remaining were dipterocarp forests. In 1991/1992 the number of fragmented forests (12) was reduced by 47.8%. Two of the fragmented forests were identified as peat swamp forest, seven dipterocarp forest and the other three was mixed of dipterocarp forests and plantation forests. Fragmentation of both dipterocarp and peat swamp forests occurred profoundly during the period between 1971/1972 and 1981/1982, which consequently increased the number of fragmented forests compared with before the period of 1971/1972 where fragmentation happened only at dipterocarp forests. However, many fragmented forests vanished between the 1981/1982 and 1991/1992 periods. PMID- 12765271 TI - Landscape ecological assessment and eco-tourism development in the South Dongting Lake Wetland, China. AB - As an important resource and the living environment of mankind, wetland has become gradually a highlight, strongly concerned and intensively studied by scientists and sociologists. The governments in the world and the whole society have been paying more and more attention on it. The Dongting Lake of China is regarded as an internationally important wetland. For a rational development and protection of the wetland, an investigation and studied on its resources and its value to tourism in the South Dongting Lake was conducted, to create an assessment system of the ecological landscapes, and to evaluate qualitatively and quantitatively the value of wetland landscape to the ecotourism. The results showed that the scenic value of the South Dongting Lake Wetland satisfied the criterion of AAAA grade of China national scenic attraction. The eco-tourism value of the landscape cultures in the South Dongting Lake Wetland was discussed with emphasis. It were formulated that a principle and frame of sustainable exploitation of the wetland landscapes and it was proposed as well that establishing a Wetland Park and developing eco-tourism in the South Dongting Lake Wetland is a fragile ecosystem with low resistance to the impact of the exploitation. Thus, we must pay intensively attention to the influence of exploitation on the landscape, take the ecological risk in account to employ a right countermeasure and avoid the negative affection. PMID- 12765272 TI - Hydrological setting of infertile species-rich wetland--a case study in the warm temperate Japan. AB - The detailed groundwater flow and water chemistry to illustrate landscape structure of the infertile peatless mire by using piezometers and groundwater wells were measured. The instruments were installed in lines through a small spring-fed wetland underlying little peat from the hillslope to the valley bottom in southwestern Japan. Flow net and EC data clearly indicated that the wetland was situated in a high-EC groundwater upspring area. The low-productivity graminous vegetation was related with four hydrological factors such as: (1) high water level; (2) low-EC (< 25 micro S/cm) groundwater; (3) weakly upward hydraulic gradient; and (4) overflowing of negatively pressured groundwater. In other words, the "old or deep groundwater" constructed the foundation of slope wetland, and maintained the high groundwater level. In contrast, overflowing "youthful groundwater" is supplied from head of slope-wetland preferentially through the shallow substratum. The plant communities of the peatless mire in southwestern Japan are similar to those of raised bog in northern cool temperate Japan. There have been some reports verifying that the underlying mineral substrata of such wetlands were quartzile rocks such as granite, rhyolite, chart and well-leached sand. Results showed (1) low cation availability affect the water acidity; (2) upward seepage of high-EC groundwater composed the foundation of the investigated peatless mire; and (3) the poor mineral condition seems to play a similar role to northern ombrotrophic (rain-fed) condition. PMID- 12765273 TI - Conservation for the landscape ecological diversity in Wulingyuan scenic area of China. AB - Wulingyuan is located at the mountainous area of the middle reach of the Yangtze River, it is one of the three nature heritages in China which ranks in the "List of World's Heritage" by UNESCO. It is characterized by quartz sandstone peaks landform with several landform components (pattern, corridor) and rich in landscape ecological diversity and biodiversity. The main patterns (ecosystem) include mid-height mountain peaks, rift-valley and streams among peaks, peaks and gullies on slopes, square mountain-platforms and peaks among blind valleys and so on. The corridor system consists of natural corridors and artificial corridors among which the stream corridors account for a major part. The fracturing of habitat is unfavorable for the biodiversity conservation, but meanwhile the habitat diversity leads to an increase in biodiversity. Therefore, it is still rich in landscape ecological diversity in Wulingyuan. The biodiversity at the level of landscape component (ecosystem) and the function of the Wulingyuan complex ecosystem, and the measures for the biodiversity conservation in Wulingyuan ecotourism area are discussed in this paper. PMID- 12765274 TI - Raman crystal lasers in the visible and near-infrared. AB - Raman lasers based on potassium gadolinium tungstate and lead tungstate crystals pumped by a to approximately 120 ps Nd: YAG laser at 1.064 microm were developed. High reflection mirrors for the Stokes wavelength have been used to generate near infrared and eye safe spectral region of 1.15 - 1.32 microm. Second harmonic generation of the generated Raman lasers was observed. Eifficient multiple Stokes and anti-Stokes picosecond generation in 64 crystals have been shown to exhibit stimulated Raman scattering on about 700 lines covering the whole visible and near-infrared spectrum. All stimulated Raman scattering (SRS) wavelengths in the visible and near-infrared spectrum are identified and attributed to the SRS active vibration modes of these crystals. PMID- 12765275 TI - Analysis of stimulated Brillouin scattering in multi-mode fiber by numerical solution. AB - Stimulated Brillouin scattering in optical fibers is described by a theoretical model and numerical analysis. The results showed that, for an optical fiber pumped by a laser beam with ns-order-pulse width and kW-order peak-power, SBS reflectivity tends to saturate when the fiber length exceeds a limit, named "effective fiber length". Using small core-diameter and long enough fiber, the SBS reflectivity level could be raised but is limited by optical damage of the entrance surface of the fiber. Therefore, just a small dynamic range can be obtained. PMID- 12765276 TI - An adaptive strategy for controlling chaotic system. AB - This paper presents an adaptive strategy for controlling chaotic systems. By employing the phase space reconstruction technique in nonlinear dynamical systems theory, the proposed strategy transforms the nonlinear system into canonical form, and employs a nonlinear observer to estimate the uncertainties and disturbances of the nonlinear system, and then establishes a state-error-like feedback law. The developed control scheme allows chaos control in spite of modeling errors and parametric variations. The effectiveness of the proposed approach has been demonstrated through its applications to two well-known chaotic systems: Duffing oscillator and Rossler chaos. PMID- 12765277 TI - Virtual local target method for avoiding local minimum in potential field based robot navigation. AB - A novel robot navigation algorithm with global path generation capability is presented. Local minimum is a most intractable but is an encountered frequently problem in potential field based robot navigation. Through appointing appropriately some virtual local targets on the journey, it can be solved effectively. The key concept employed in this algorithm are the rules that govern when and how to appoint these virtual local targets. When the robot finds itself in danger of local minimum, a virtual local target is appointed to replace the global goal temporarily according to the rules. After the virtual target is reached, the robot continues on its journey by heading towards the global goal. The algorithm prevents the robot from running into local minima anymore. Simulation results showed that it is very effective in complex obstacle environments. PMID- 12765278 TI - Application of stochastic method to optimum design of energy-efficient induction motors with a target of LCC. AB - For an energy-efficient induction machine, the life-cycle cost (LCC) usually is the most important index to the consumer. With this target, the optimization design of a motor is a complex nonlinear problem with constraints. To solve the problem, the authors introduce a united random algorithm. At first, the problem is divided into two parts, the optimal rotor slots and the optimization of other dimensions. Before optimizing the rotor slots with genetic algorithm (GA), the second part is solved with TABU algorithm to simplify the problem. The numerical results showed that this method is better than the method using a traditional algorithm. PMID- 12765279 TI - Analysis and design of DSP-based dual-loop controlled UPS inverters. AB - This paper presents a novel digital dual-loop control scheme of the PWM (Pulse width modulate) inverter. Deadbeat control technique are employed to enhance the performance. Half switching period delayed sampling and control tining strategy is used to improve the system dynamic response. Simulation and experimental results presented in the paper verified the validity of the proposed control scheme. PMID- 12765280 TI - Noise and linearity optimization methods for a 1.9GHz low noise amplifier. AB - Noise and linearity performances are critical characteristics for radio frequency integrated circuits (RFICs), especially for low noise amplifiers (LNAs). In this paper, a detailed analysis of noise and linearity for the cascode architecture, a widely used circuit structure in LNA designs, is presented. The noise and the linearity improvement techniques for cascode structures are also developed and have been proven by computer simulating experiments. Theoretical analysis and simulation results showed that, for cascode structure LNAs, the first metallic oxide semiconductor field effect transistor (MOSFET) dominates the noise performance of the LNA, while the second MOSFET contributes more to the linearity. A conclusion is thus obtained that the first and second MOSFET of the LNA can be designed to optimize the noise performance and the linearity performance separately, without trade-offs. The 1.9GHz Complementary Metal-Oxide Semiconductor (CMOS) LNA simulation results are also given as an application of the developed theory. PMID- 12765282 TI - A new algorithm of brain volume contours segmentation. AB - This paper explores brain CT slices segmentation technique and some related problems, including contours segmentation algorithms, edge detector, algorithm evaluation and experimental results. This article describes a method for contour baed segmentation of anatomical structures in 3D medical data sets. With this method, the user manually traces one or more 2D contours of an anatomical structure of interest on parallel planes arbitrarily cutting the data set. The experimental results show the segmentation based on 3D brain volume and 2D CT slices. The main creative contributions in this paper are: (1) contours segmentation algorithm; (2) edge detector; (3) algorithm evaluation. PMID- 12765281 TI - Building a highly available and intrusion tolerant Database Security and Protection System (DSPS). AB - Database Security and Protection System (DSPS) is a security platform for fighting malicious DBMS. The security and performance are critical to DSPS. The authors suggested a key management scheme by combining the server group structure to improve availability and the key distribution structure needed by proactive security. This paper detailed the implementation of proactive security in DSPS. After thorough performance analysis, the authors concluded that the performance difference between the replicated mechanism and proactive mechanism becomes smaller and smaller with increasing number of concurrent connections; and that proactive security is very useful and practical for large, critical applications. PMID- 12765283 TI - Predictive functional control (PFC) and its application in chlorinated polyethylene process. AB - The main principle and the characteristic of Predictive Functional Control (PFC) strategy are presented in this paper and the corresponding control system aid design software APC-PFC is also introduced. For a chlorinated polyethylene (CPE) process, a design scheme of cascade predictive functional control system is described and the control performance is improved obviously. PMID- 12765284 TI - System design and analysis of the trans-critical carbon-dioxide automotive air conditioning system. AB - As an environmentally harmless and feasible alternate refrigerant, CO2 has attracted worldwide attention, especially in the area of automobile air conditioning (AAC). The thermal property of CO2 and its trans-critical refrigeration cycle is very different from that of the traditional CFC or HCFC system. The detailed process of CO2 system thermal cycle design and optimization is described in this paper. System prototype and performance test bench were developed to analyze the performance of the CO2 AAC system. PMID- 12765285 TI - Three-dimensional transient numerical simulation for intake process in the engine intake port-valve-cylinder system. AB - This paper presents a KIVA-3 code based numerical model for three-dimensional transient intake flow in the intake port-valve-cylinder system of internal combustion engine using body-fitted technique, which can be used in numerical study on internal combustion engine with vertical and inclined valves, and has higher calculation precision. A numerical simulation (on the intake process of a two-valve engine with a semi-sphere combustion chamber and a radial intake port) is provided for analysis of the velocity field and pressure field of different plane at different crank angles. The results revealed the formation of the tumble motion, the evolution of flow field parameters and the variation of tumble ratios as important information for the design of engine intake system. PMID- 12765286 TI - Geometrical nonlinear stability analyses of cable-truss domes. AB - The nonlinear finite element method is used to analyze the geometrical nonlinear stability of cable-truss domes with different cable distributions. The results indicate that the critical load increases evidently when cables, especially diagonal cables, are distributed in the structure. The critical loads of the structure at different rise-span ratios are also discussed in this paper. It was shown that the effect of the tensional cable is more evident at small rise-span ratio. The buckling of the structure is characterized by a global collapse at small rise-span ratio; that the torsional buckling of the radial truss occurs at big rise-span ratio; and that at proper rise-span ratio, the global collapse and the lateral buckling of the truss occur nearly simultaneously. PMID- 12765288 TI - Cloning and characterization of a glucose 6-phosphate/phosphate translocator from Oryza sativa. AB - Plastids of nongreen tissues import carbon as a source of biosynthetic pathways and energy, and glucose 6-phosphate is the preferred hexose phosphate taken up by nongreen plastids. A cDNA clone encoding glucose 6-phosphate/phosphate translocator (GPT) was isolated from a cDNA library of immature seeds of rice and named as OsGPT. The cDNA has one uninterrupted open reading frame encoding a 42 kDa polypeptide possessing transit peptide consisting of 70 amino acid residues. The OsGPT gene maps on chromosome 8 of rice and is linked to the quantitative trait locus for 1000-grain weight. The expression of OsCPT is mainly restricted to heterotrophic tissues. These results suggest that glucose 6-phosphate imported via GPT can be used for starch biosynthesis in rice nongreen plastids. PMID- 12765287 TI - Geotechnical behavior of the MSW in Tianziling landfill. AB - The valley shaped Tianziling landfill of Hangzhou in China built in 1991 to dispose of municipal solid waste (MSW) was designed for a service life of 13 years. The problem of waste landfill slope stability and expansion must be considered from the geotechnical engineering point of view, for which purpose, it is necessary to understand the geotechnical properties of the MSW in the landfill, some of whose physical properties were measured by common geotechnical tests, such as those on unit weight, water content, organic matter content, specific gravity, coefficient of permeability, compressibility, etc. The mechanical properties were studied by direct shear test, triaxial compression test, and static and dynamic penetration tests. Some strength parameters for engineering analysis were obtained. PMID- 12765289 TI - Emergence of a new satellite RNA from cucumber mosaic virus isolate P1. AB - The cucumber mosaic virus (CMV) isolate P1 caused very mild symptoms on many plant species. After serial passages by mechanical inoculation over five years, CMV P1 caused severe symptoms on several tobacco cultivars and tomato. A specific band of approximately 0.3 kb in length was amplified by RT-PCR with primers synthesized based on reported CMV satellite RNA (satRNA) sequences. Sequence analysis showed there were two satRNAs (Sat-P1-1 and Sat-P1-2). Sat-P1-1 contained 335 nucleotides, and Sat-P1-2 contained 394 nucleotides. These two satRNAs shared 64% overall nucleotide sequence homology, and differences between the two satRNAs included mutations as well as deletions. Sat-P1-1 was identical to a satRNA (Z96099) reported in 1995 in CMV P1. Based on differences in the sequence and secondary structure between these two satRNAs, we conclude that Sat P1-2 represents the emergence of a new satellite (necrotic satellite) from attenuated satRNA populations. The possible effect of the emergence of this new satRNA is discussed. PMID- 12765290 TI - Expression of a fusion protein of human ciliary neurotrophic factor and soluble CNTF-receptor and identification of its activity. AB - Ciliary neurotrophic factor (CNTF) has pleiotropic actions on many neuronal populations as well as on glia. Signal transduction by CNTF requires that it bind first to CNTF-R, permitting the recruitment of gp130 and LIF-R, forming a tripartite receptor complex. Cells that only express gp130 and LIF-R, but not CNTF-R are refractory to stimulation by CNTF. On many target cells CNTF only acts in the presence of its specific agonistic soluble receptors. We engineered a soluble fusion protein by linking the COOH-terminus of sCNTF-R to the NH2 terminus of CNTF. Recombinant CNTF/sCNTF-R fusion protein (Hyper-CNTF) was successfully expressed in COS-7 cells. The apparent molecular mass of the Hyper CNTF protein was estimated from western blots to be 75 kDa. Proliferation assays of transfected BAF/3 cells in response to CNTF and Hyper-CNTF were used to verify the activity of the cytokines. The proliferative results confirmed that CNTF required homodimerization of the gp130, CNTF-R and LIF-R receptor subunit whereas Hyper-CNTF required heterodimerization of the gp130 and LIF-R receptor subunit. We concluded that the fusion protein Hyper-CNTF had superagonistic activity on target cells expressing gp130 and LIF-R, but lacking membrane-bound CNTF-R. PMID- 12765291 TI - Agrobacterium-mediated transformation of herbicide resistance in creeping bentgrass and colonial bentgrass. AB - Embryogenic calli were induced from the seeds of creeping bentgrass (Agrostis palustris Huds.) cv. Regent and colonial bentgrass (Agrostis Tenuis Sibth. Fl. Oxen.) cv. Tiger. The embryogenic calli were precultured on fresh medium for 4-7 days and then co-cultivated with Agrobacterium tumefaciens, LBA4404, which contains plasmid vector-pSBGM harboring bar coding region, synthetic green fluorescent protein (sGFP) coding region and matrix attachment region (MAR). After 3 days of co-cultivation, the calli were washed thoroughly and transferred to MS medium containing 2 mg/L of 2, 4-D, 12-15 mg/L phosphinothricin (PPT) and 250 mg/L of cefotaxime. After 2-3 months of selection, the actively growing calli of 'Regent' and 'Tiger' were transferred to MS medium with 12-15 mg/L PPT and 250 mg/L cefotaxime for regeneration. The putative transformants were maintained on MS medium with 3 mg/L PPT for long period but control died within 1 month. After establishing in greenhouse, the transformants also showed strong resistance to 0.4% of herbicide Basta but control plants died within 2 weeks. Under confocal microscope, both young leaves and roots showed significant GFP expression. PCR analysis revealed the presence of a DNA fragment of GFP gene at the expected size (380 bp) in the transformants and its absence in a randomly selected control plant. PMID- 12765292 TI - CH4 emission and recovery from municipal solid waste in China. AB - Methane ( CH4) is an important greenhouse gas and a major environmental pollutant, second only to carbon dioxide (CO2) in its contribution to potential global warming. In many cases, methane emission from landfills otherwise emitted to the atmosphere can be removed and utilized, or significantly reduced in quantity by using coat-effective management methods. The gas can also be used as a residential, commercial, or industrial fuel. Therefore, emission reduction strategies have the potential to become low cost, or even profitable. The annual growth rate of Municipal Solid Waste (MSW) output in China is 6.24%, with the highest levels found in South China, Southwest China and East China. Cities and towns are developing quickly in these regions. MSW output was only 76.36 Mt in 1991 and increased to 109.82 Mt in 1997, registering an average increase of 43.8% . In China, methane emission from landfills also increased from 5.88 Mt in 1991 to 8.46 Mt in 1997; so the recovery of methane from landfills is a profitable project. PMID- 12765293 TI - Of pH and Eh on release of nitrogen and phosphorus from sediments of West Lake. AB - The effects of pH and Eh on release of nitrogen and phosphorus from sediments of West Lake under different conditions were investigated by simulation test. Results showed that the net flux of NH4(+) -N release from sediments increased with pH, but NO3(-) -N showed negative-going release at all tested pH levels. The net flux of NH4(+) -N release from sediments was higher under anaerobic or aerobic condition of the overlying water, but only under aerobic condition would net release of NO3(-) -N occur. It was also shown that phosphorus released was mainly in its inorganic form, higher pH and anaerobic conditions of overlying water greatly stimulated release of phosphorus. In situ measurement at several West Lake locations indicated that sediment resuspension induced by boat propeller stimulated nutrients release from sediment into overlying water. PMID- 12765294 TI - Study on the thermal decomposition kinetics of nano-sized calcium carbonate. AB - This study of the thermal decomposition kinetics of various average diameter nano particles of calcium carbonate by means of TG-DTA ( thermogravimetry and differential thermal analysis) showed that the thermal decomposition kinetic mechanisms of the same crystal type of calcium carbonate samples do not vary with decreasing of their average diameters; their pseudo-active energy (a); and that the top-temperature of decomposition T(p) decreases gently in the scope of micron sized diameter, but decreases sharply when the average diameter decreases from micron region to nanometer region. The extraordinary properties of nano-particles were explored by comparing the varying regularity of the mechanisms and kinetic parameters of the solid-phase reactions as well as their structural characterization with the variation of average diameters of particles. These show that the aggregation, surface effect as well as internal aberrance and stress of the nano-particles are the main reason causing both E(a) and T(p) to decline sharply with the decrease of the average diameter of nano-particles. PMID- 12765295 TI - Polymorphisms in the genes for coagulation factor II, V, VII in patients undergoing coronary angiography. AB - OBJECTIVE: To determine whether polymorphisms in the genes for coagulation factor II, V, VII could predispose an individual to increase risk for coronary artery disease (CAD) and/or myocardial infarction (MI) in Chinese. METHODS: We screened coagulation factor II(G20210A),V(G1691A),VII (R353Q and HVR4) genotype in 374 patients undergoing coronary angiography by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) assay. RESULTS: The R353Q and HVR4 genotype of the factor VII distribution was in accordance with Hardy Weinberg equilibrium. The frequencies of FVII genotype or allele did not show statistically significant differences between CAD group and controls or between male and female. The frequencies of the Q allele and (RQ + QQ) genotype were significantly higher among the CAD patients without myocardial infarction (MI) history than among those with MI history (P < 0.05). However, HVR4 polymorphism was not significantly different within groups. We only find one normal control of factor II (G20210A) mutation. No coagulation factor V(G1691A) mutation was found in the CAD patients and controls. CONCLUSION: The factor II(G20210A) ,V(G1691A) mutation is absent and may not be a major genetic factor for CAD and/or MI; the Q allele of the R353Q polymorphism of the factor VII gene may be a protective genetic factor against myocardial infarction in Chinese. PMID- 12765296 TI - Comparison study of harmonic imaging (HI) and fundamental imaging (FI) in fetal echocardiography. AB - OBJECTIVES: To directly compare the quality of harmonic imaging (HI) and fundamental imaging (FI) in fetal echocardiography and to determine any differences in image quality between the two modalities. METHODS: Fetal echocardiograms were performed with the use of FI and HI in 58 fetuses, image quality and visualization of left and right atria, left and right ventricles, mitral and tricuspid valves, aortic and pulmonary valves, left and right ventricular outflow tracts were evaluated and compared between FI and HI. RESULTS: Mean HI scores were higher than mean FI scores (2.73 +/- 0.43 vs 2.16 +/ 0.69, P < 0.001) for all the cardiovascular structures evaluated. Compared with FI, HI improved the image quality and visualization of fetal cardiac structures in subjects with both good (2.73 +/- 0.43 vs 2.88 +/- 0.32, P < 0.001) and suboptimal (1. 65 +/- 0.41 vs 2.58 +/- 0.47, P < 0.001) echocardiographic windows. The interobserver correlation coefficient for the grading scores was 0.74 (P < 0.001). CONCLUSIONS: harmonic imaging enhances and improves the image quality of fetal echocardiography; and has important potential role in cardiac imaging in the fetus. PMID- 12765297 TI - Autoimmunity and geriatrics: clinical significance of autoimmune manifestations in the elderly. AB - The immune system undergoes continuous morphologic and functional changes throughout the years, and it is now believed that the immune response has its peak function in puberty and gradually decreases with age (immunosenescence). Recent studies in healthy octogenarian patients suggest that the immune system, instead of suffering a generalized deterioration, undergoes a remodelling/readjustment of its major functions. Increase in two contrasting phenomena coexist in immunosenescence: on the one hand, a decrease in the capacity of the immune response and, on the other hand autoantibody production. The possible consequences of this progressive 'ageing' of the immune system are the increase in autoimmune phenomena, incidence of neoplasia and predisposition to infections. The study of autoimmune manifestations in elderly populations should be considered a priority for future medical research because of increasing life expectancy, especially in developed countries. This review analyses the main immune disorders associated with immunosenescence, the prevalence and clinical significance of autoantibodies in the elderly and the clinical expression of the main autoimmune diseases in older patients. PMID- 12765298 TI - Assessment of Be1 and Be2 cells in systemic lupus erythematosus indicates elevated interleukin-10 producing CD5+ B cells. AB - Recent studies indicate that normal B cells can be primed to differentiate into two distinct cytokine-secreting effector subsets, Be1 and Be2. The aim of this study was to analyse, for the first time, Be1 and Be2 cells at the single cell level in SLE patients using the recently developed technique of flow cytometry for intracellular cytokines. Peripheral blood mononuclear cells (PBMC) from SLE patients and age- and sex-matched normal controls were cultured for 24 h in the presence or absence of phorbal myristate acetate and ionomycin (PMA/I) or lipopolysaccharide (LPS). The production of type I (IFN-gamma, IL-2) and type 2 (IL-4, IL-5, IL-6, IL-10, IL-13) cytokines by B cells (and IL-10 production by fractionated CD5+ and CD5- B cells) was investigated using an intracellular cytokine staining technique and flow cytometry. In the absence of PMA/I stimulation, the percentage of B cells from SLE patients was significantly lower than those of normal subjects and significantly more SLE B cells spontaneously produced IL-10 than controls. Moreover, CD5+ B cells from SLE patients were enriched for cells with signs of previous in vivo activation and for high levels of IL-10 production. A significant positive correlation was observed between the percentage of IL-10- and IL-6-producing PMA/I-stimulated B cells in SLE patients, but not in controls. There were no significant differences in the production of other cytokines by B cells of SLE patients and normal subjects. In conclusion, a general alteration of type 1 and type 2 cytokine production by B cells is not observed in SLE patients. The role of B cell cytokines in the pathogenesis of SLE appears to be exerted by elevated secretion of in vivo IL-10, which may play an important role in the immune dysregulation observed in SLE patients. Moreover, the cross regulation of IL-10 and IL-6 is disrupted in SLE patients. PMID- 12765299 TI - Anti-prothrombin antibodies cause thrombosis in a novel qualitative ex-vivo animal model. AB - Anti-prothrombin antibodies (aPT) are associated with thrombotic manifestations, and their association with reproductive failure is debatable. The aim of this study was to examine whether aPT could induce thrombosis and other clinical manifestations of the anti-phospholipid syndrome (APS). Mice were immunized with either prothrombin, beta2-glycoprotein-I (beta2GPI), or beta2GPI followed by prothrombin. The presence of clinical manifestation of APS, including thrombocytopenia, lupus anticoagulant and fetal resorption rates, was evaluated in all mice groups compared with nonimmunized mice. Thrombosis was studied in a novel ex-vivo model in which the aorta was sutured for 1 min and the presence or absence of visible thrombus was qualitatively evaluated. Immunized mice developed high autoantibody levels directed towards their immunizing autoantigens. The groups immunized with beta2GPI or beta2GPI/prothrombin, but not with prothrombin alone, developed prolonged aPTT, thrombocytopenia and increased fetal resorption rate. All prothrombin-immunized mice as well as most beta2GPI/prothrombin immunized mice developed visible thrombus within the aorta. Some beta2GPI immunized mice developed very mild thrombus. None of the CFA/PBS-injected or the nonimmunized mice developed such thrombus. Active immunization with prothrombin or beta2GPI/prothrombin is associated with prothrombotic activity of blood in an ex-vivo model. This is the first direct evidence for thrombus induction by aPT. PMID- 12765300 TI - Risk-attitude and patient treatment preferences. AB - Women's treatment preferences for lupus nephritis vary widely even after adjusting for sociodemographic factors and disease severity. Attitude toward risk may partially explain interpatient variability in treatment preference. The objective of this study was to examine the association between 'risk-attitude' and patient treatment preferences in lupus nephritis. Sixty-five premenopausal women with systemic lupus erythematosus were interviewed. Patient preferences for cyclophosphamide versus azathioprine for the treatment of lupus nephritis were ascertained using an Adaptive Conjoint Analysis questionnaire. Risk-attitude was ascertained by asking patients to choose between a pair of lotteries having the same expected value but differing in spread (the difference between the worst and best outcomes). Respondents preferring the wider spread were classified as relatively more risk-seeking and those preferring the narrower spread were classified as relatively more risk-averse. Twenty-eight percent of respondents were classified as relatively more risk-seeking. Risk-seeking women were more likely to prefer cyclophosphamide for the treatment of lupus nephritis compared with risk-averse women [least square mean (+/- SD) preference for cyclophosphamide 63 +/- 3 among risk-seeking women versus 55 +/- 2 among risk averse women (P < 0.03)]. The association between risk attitude and treatment preference persisted asthe probabilities of adverse events were varied to reflect the range of risks reported in the literature. Our results suggest that patients' relative risk-attitudes, as measured by a lottery task, are related to their treatment preferences. Differences in risk-attitude may help explain the inter patient variability in treatment preferences. PMID- 12765301 TI - Clinical, immunogenetic and outcome features of Hispanic systemic lupus erythematosus patients of different ethnic ancestry. AB - The aim of this study was to compare and contrast the clinical, immunogenetic and outcome features of two subgroups of Hispanic patients with systemic lupus erythematosus (SLE), one from Northern Spain (Spaniards) and one of from the USA (Hispano-Americans: Hispanics primarily of Mexican ancestry (Amerindian and Spaniard backgrounds). Patients with SLE as per the American College of Rheumatology classification criteria, from two University-affiliated Hospitals (Universidad de Cantabria) and disease of five or less years in duration (n = 28) and with four years of follow up constituted the Spaniard subgroup. Fifty-two patients of Hispano-American ancestry from the LUMINA (Lupus in Minority populations: Nature versus Nurture) cohort constituted the Hispano-American subgroup. Patients were studied using a similar protocol. In short, sociodemographic, clinical, immunological, immunogenetic and psychosocial and behavioral features were obtained at enrollment into the study (baseline visit) and yearly thereafter. The relationship between these variables and disease activity at baseline and over time, as measured by the systemic lupus activity measure (SLAM) and disease damage, as measured by the SLICC (Systemic Lupus International Collaborating Clinics) Damage Index (SDI) were determined. Variables found to be significant at P = 0.10 were then entered into multivariable linear regression models with disease activity at baseline and over time, and damage as the outcome measures. Patients of Hispano-American and Spaniard ethnicity had comparable sociodemographic features except for home density, which was higher among the Hispano-Americans. HLA-DRB1*08 was associated with SLE among the Hispano-Americans but not among the Spaniards. Hispano American patients had more severe disease as manifested by more frequent clinical manifestations (renal and neurological), higher SLAM scores at baseline and over time and higher SDI scores at the year 4 visit (that despite the fact that Hispano-American patients had overall shorter disease duration than the Spaniard patients). Hispano-American ethnicity, younger age at disease onset and the number of ACR criteria at baseline and over time were consistently associated with disease activity, whereas increased home density and the absence of HLA DRB1*0301 were significant predictors only over time. Disease damage was associated with disease activity over time, the number of ACR criteria at baseline, increased home density and the presence of HLA-DRB1*08. This is the first longitudinal study of SLE in two different Hispanic subgroups. Hispanics with a strong Amerindian background have a more serious disease than that observed in Spaniards. Genetic and socio-economic differences between these two Hispanic subgroups probably account for these findings. PMID- 12765302 TI - Immunoregulatory defects in patients with systemic lupus erythematosus in clinical remission. AB - Little is known about the immune system of patients with systemic lupus erythematosus (SLE) during periods of silent disease. To address this issue we analysed lymphoid populations andcytokine production of mononuclear cells obtained from SLE patients in remission. We studied 43 patients with inactive disease, 10 with active disease and 30 controls. Remission was defined as at least 1 year during which lack of clinical disease activity permitted withdrawal of all treatment. Remission length ranged from 1 to 30 years. Flow cytometry and ELISA were used to study lymphoid populations (CD4, CD8 and CD19) and cytokine production (IL-2, 4, 10, 12 and 18). Patients with short remission periods (up to 15 years) exhibited an increased percentage of B cells; production of IL-2, IL-10 and IL-12 was decreased; production of IL-18 was increased. Interestingly, patients from groups with long time of inactive disease had corrected most alterations, but had an impaired IL-18 expression. IL-12 production correlated strongly with the length of the remission period (r = 0.7565). The immune system of patients with inactive lupus has partially corrected the disturbances present during disease activity. This is accomplished gradually, sometimes until counter regulatory alterations are developed. This may allow patients to remain without disease activity. PMID- 12765303 TI - The prevalence of coeliac disease antibodies in patients with the antiphospholipid syndrome. AB - High prevalence of coeliac disease (CD) has been reported in various autoimmune disorders, buthas not been studied in the antiphospholipid syndrome (APS). We aimed to establish the prevalence of CD antibodies in a cohort of APS patients, and to examine whether CD may be responsible for some of the manifestations of APS. Fifty-seven patients (47 females, 10 males) with APS were studied for clinical manifestations and serological markers of the disease, as well as the presence of anti-endomysial antibodies using an ELISA assay (EMA-ELISA). Control subjects were 171 healthy individuals, age- and sex-matched (141 females). Eight patients with APS (14%, six females) were found to have EMA-ELISA antibodies, compared with 2/141 (1.1%) of controls (P = 0.0003). Antibodies against beta2 glycoprotein-I (beta2GPI) epitopes (GRTCPKPDDLP) were more prevalent in EMA positive patients than in EMA-negative patients (P = 0.006). Vasculitic skin lesions were significantly more common in EMA-ELISA-positive compared with EMA ELISA-negative patients(62.5 versus 16.3%, P = 0.01). Among the skin manifestations, superficial cutaneous necrosis (37.5 versus 2%, P = 0.007) was more prevalent in EMA-ELISA-positive than in EMA-ELISA-negative patients. EMA ELISA antibodies are common in APS, and their presence is associated with high prevalence of antibodies recognizing certain beta2-glycoprotein epitopes, and with cutaneous manifestations of APS. PMID- 12765304 TI - Association of homozygous deletion of the Humhv3005 and the VH3-30.3 genes with renal involvement in systemic lupus erythematosus. AB - To investigate whether deletion of the Humhv3005 and the homologous VH3-30.3 (both share an identical amino acid sequence) genes is associated with susceptibility and/or certain clinical manifestations of systemic lupus erythematosus (SLE), DNA from 108 Korean SLE patients and 102 healthy subjects were analysed for the status of hv3005 gene by polymerase chain reaction-enzyme linked immunosorbent assay. This method consists of amplification of selected germline VH3 genes with biotinylated primers, efficient capture of amplicons onto streptavidin-coated wells, and quantitative typing of bound VH3 gene with diagnostic oligonucleotides. We found that deletion of the hv3005 gene (including VH3-30.3) was more frequent in SLE patients than in healthy controls (26.9 versus 11.8%, P = 0.006, odds ratio 2.75). When clinical features were examined, patients with hv3005 deletion have a higher frequency of lupus nephritis (LN) (75.9 versus 44.3% for those without, P = 0.004), and higher activity index [median (range), 6 (2-14) versus 4 (1-16) for those without, P = 0.044] when biopsy-proven LN was studied. Collectively, our data suggest that deletion of the hv3005 and the 3-30.3 genes may predispose individual SLE patients to the development of LN. PMID- 12765305 TI - Factor V Leiden mutation in Sneddon syndrome. AB - Sneddon syndrome (SNS) is characterized by the association of ischaemic cerebrovascular events and widespread livedo racemosa. Its pathophysiology is still controversial. The aim of this study was to evaluate the prevalence of factor V Leiden mutation in consecutive patients referred for SNS according to antiphospholipid antibodies (aPL) status. Fifty-three Caucasian patients were enrolled from 1996 to 2001. Diagnosis of SNS was based on the presence of a widespread livedo racemosa and at least one clinical neurologic ischaemic event. The following investigations were performed: detection of antithrombin III, protein C and protein S deficiency, lupus anticoagulant, anticardiolipin and anti beta2 glycoprotein I antibodies, biologic false-positive test for syphilis, and factor V Leiden mutation by direct genomic analysis. Fisher's test and t-test were used for statistics. Detection of aPL on multiple determinations was negative in 31 patients (group 1) and positive in 22 patients (group 2). Factor V Leiden mutation was detected in six patients (11.3%), heterozygous in all. The frequency of this mutation was statistically higher in group 1 (6/31, 19.3%) than in group 2 (0/22; P = 0.035). Within aPL-negative SNS, the comparison of patients with versus without factor V Leiden mutation showed no difference for clinical data or familial history of thrombosis. A high prevalence of heterozygous factor V mutation was found in aPL-negative patients with SNS. This finding adds further arguments to consider SNS as a heterogeneous entity. PMID- 12765306 TI - Drug-induced lupus-like syndrome associated with severe autoimmune hepatitis. AB - Atorvastatin and other members of the statin family are widely used for the treatment of hypercholesterolaemia in order to reduce the risk of atherosclerosis and cardiovascular disease. Atorvastatin-induced adverse events are mostly mild and only a few cases of lupus-like syndrome or severe acute hepatitis have been documented. In this case report we describe a patient who developed an atorvastatin-induced severe autoimmune hepatitis. In addition, this patient presented with a concomitant systemic lupus-like syndrome which has been already described for statins but not in association with severe liver disease. Although the drug was immediately withdrawn the disease persisted and even deteriorated to a fulminant disease with evidence of acute hepatic failure. The patient failed to respond to conventional immunosuppression with corticosteroids and azathioprine. Only the introduction of intense immunosuppressive therapy, as used in solid organ transplantation, led to a complete and sustained recovery of the patient. Interestingly, the patient was HLA DR3- and HLA DR4-positive, which are well known genetic factors associated with autoimmune diseases. This case is the first report of a drug-induced lupus-likesyndrome concomitant with a severe autoimmune hepatitis in a genetically predisposed patient. PMID- 12765307 TI - Atypical cutaneous findings in a patient with systemic lupus erythematosus. AB - A previously diagnosed systemic lupus erythematosus patient presented with arthralgia, skin rash and muscular weakness. When treated with high-dose corticosteroids and methotrexate she improved, except for a persistent lesion in the hand which evolved into a profound ulcer, along with tender subcutaneous nodules in the calf. A skin biopsy disclosed necrotizing vasculitis with giant cell granuloma revealing acid fast positive bacteria on ziels nilsen staining. A chest X-ray disclosed miliary tuberculosis (TB). The patient was diagnosed as miliary TB with prominent cutaneous involvement and treated with four anti tuberculous drugs with slow resolution of her systemic, pulmonary and skin signs. PMID- 12765308 TI - A fatal case of systemic lupus erythematosus complicated by acute pancreatitis, invasive aspergillosis and features of thrombotic thrombocytopenic purpura. AB - We describe the case of a 23-year-old woman with a mild form of systemic lupus erythematosus who presented a febrile illness rapidly followed by general worsening, neurologic involvement, renal failure and coma. While hospitalized in the intensive care unit she also suffered from acute pancreatitis, microangiopathic hemolitic anemia, thrombocytopenia and prolongation of clotting times. Despite aggressive treatment the patient died at day 17 of hospitalization in the intensive care unit. At autopsy necrotico-hemorragic pancreatitis, diffuse pneumonia, peritonitis and cerebral edema were present. Most striking was the presence of invasive aspergillosis, which was detected in all organs examined. In this case thrombotic thrombocytopenic purpura, invasive aspergillosis and multiorgan failure including acute pancreatitis were present. The relationship between the three entities is complex, and it is difficult to establish which of the different events took place first and triggered the others. PMID- 12765309 TI - The differential diagnosis of central nervous system lupus has expanded. PMID- 12765310 TI - Insulin-like growth factor-II gene ApaI polymorphism was not associated with systemic lupus erythematosus. PMID- 12765311 TI - Inhaled corticosteroids for asthma: common clinical quandaries. AB - This narrative review provides evidence-based explanations to some of the common clinical concerns regarding inhaled corticosteroids. Inhaled corticosteroids are the treatment of choice for a newly diagnosed asthmatic patient. Better results are obtained when treatment is initiated as soon as the diagnosis is made. Asthma control can be achieved and maintained in most patients with a low or moderate dose of inhaled corticosteroid administered in two daily doses. Longer duration of treatment provides more sustained benefits than treatment that is intermittent and for short periods of time. The clinical benefits can be observed within 24 hours of commencing treatment and may be more pronounced in patients with an eosinophilic bronchitis. Inhaled corticosteroids provide additional benefit when used in conjunction with prednisone in acute severe asthma. Low doses do not have clinically deleterious side effects on the bones, growth, eye, or hypothalamo pituitary-adrenal-axis. However, they do not normalize lung function and prevent structural changes in the airway wall in all asthmatic patients. PMID- 12765312 TI - Duration of deep inspiration and subsequent airway constriction in vivo. AB - The effects of a deep inspiration (DI) in asthmatics differ from those observed in healthy subjects. When considering the effects of a DI, an implicit assumption is that all the airways are distending at the same rate as the lung parenchyma. However, with such rapid lung inflation, the ability of contracted airways to dynamically follow the lung parenchyma was recently shown to significantly lag the lung inflation. Another potentially important variable in the response of the individual airways to a DI that has not been well studied is the duration of the DI maneuver. The current study examines the effects of increasing duration at TLC during a DI on subsequent airway caliber. In five anesthetized and ventilated mongrel dogs, after DIs of increasing duration, changes in airway size were measured over the subsequent 5-minute period using high-resolution computed tomography. Results show that the duration of the maneuver is extremely important, leading to a qualitative change in the airway response. A long DI (> or = 30 seconds) caused subsequent airway dilation, while a shorter DI (< 30 seconds) caused bronchoconstriction. The precise mechanism underlying these observations is uncertain but seems to be related to intrinsic properties within the contracted airway smooth muscle. PMID- 12765313 TI - Asthma caused by human seminal plasma allergy. AB - Human seminal plasma allergy (HSPA) in women is a rare phenomenon and can be life threatening. Hypersensitivity reactions occur during or immediately after coitus and are usually caused by sensitization to proteins in the seminal fluid. This can be very distressing for the patients and their partners. Young, married female patients with episodic asthma dating to their marriage with a history of difficulty in conceiving were specifically asked for local and systemic symptoms chronologically related to coitus. This methodology was adopted keeping in view our social conditions and sensitive nature of the information sought. Two patients thus detected were evaluated further. Condom usage, on our advice, abolished both local and systemic postcoital symptoms. Both patients tested positive to intradermal skin test with the seminal plasma antigen (SPA) derived from their respective husbands and, in patient 1, was accompanied by intense burning, irritation, and itching of the skin. Specific IgE against SPA was demonstrated in the serum of both patients. Elevated levels of total IgE were also detected. A high index of suspicion is needed to diagnose HSPA, especially the local forms, which can easily be overlooked due to patient embarrassment and lack of physician awareness. PMID- 12765314 TI - Bridging the gap between doctors' and patients' expectations of asthma management. AB - OBJECTIVES: To assess the prevalence of asthma symptoms, their impact on daily activities, and perceptions of disease severity among people with asthma. METHODS: A telephone survey of 699 people with asthma was conducted in 1999 in metropolitan and nonmetropolitan New South Wales, Victoria, and Queensland, Australia. RESULTS: Forty-two percent of adults and 26% of children reported experiencing asthma symptoms at least every 2-3 days. Thirty-seven percent of adults and 26% of children reported using a reliever more than four times in the previous week. Of those for whom preventer therapy had been prescribed (61% of respondents), 30% of children and 45% of adults did not use their preventer as instructed. A high proportion of respondents reported avoiding physical and social activities because of their asthma, while 75% said asthma generally made them feel tired. Many respondents attributed frustration (61%), irritability (57%), fear (38%), and worry (43%) to their asthma. Only 50% of respondents had been reviewed by a general practitioner for asthma in the past year. Respondents generally underestimated the severity of their asthma, compared with symptom frequencies reported. CONCLUSIONS: The Living with Asthma Survey suggests that national asthma management goals are not being achieved in a high proportion of patients, with evidence for both underprescribing and underusage of preventer medication. Achieving closer alignment between medical and patient perspectives is an important goal of asthma education and management in order to help bridge the gap between current concepts of best practice and the reality of persistently poor asthma outcomes. PMID- 12765315 TI - Asthma and panic attacks among youth in the community. AB - OBJECTIVE: To determine the association between asthma and panic attacks among youth in the community. METHOD: Data were drawn from the Methods for the Epidemiology of Child and Adolescent Mental Disorders (MECA) Study (n = 1285), a community-based sample of youth age 9-17 in the United States. Multivariate logistic regression analyses were used to determine the association between asthma and both panic attacks and panic disorder, adjusting for differences in sociodemographic characteristics and comorbid mental disorders. RESULTS: Asthma was associated with a significantly increased likelihood of panic attacks [OR= 1.5 (1.01, 2.2)]. This effect was specific and persisted after adjusting for differences in demographics and psychiatric comorbidity. Severe asthma was associated with an even greater likelihood of panic attacks [OR = 2.2 (1.3, 4.0)]. There was a dose-response relationship between number of panic symptoms during a panic attack and the likelihood of asthma [OR = 1.2 (1.1, 1.3)] and severe asthma [OR = 1.3 (1.1, 1.4)], which remained significant after adjusting for differences in sociodemographic characteristics and comorbid mental disorders. CONCLUSIONS: These data suggest a significant association between asthma, severe asthma, and panic attacks among youth in the community. Replication of these findings is needed, as are future studies that investigate the nature of these links. In light of the increasing prevalence of asthma and hospitalization for asthma among youth in the United States and worldwide, these associations may be worthwhile to consider in future investigations. PMID- 12765316 TI - Leukotriene synthesis is increased by transcriptional up-regulation of 5 lipoxygenase, leukotriene A4 hydrolase, and leukotriene C4 synthase in asthmatic children. AB - Leukotrienes (LTs) are recognized to be important mediators in asthma. Recent studies revealed that LT synthesis is controlled by the regulation of LT synthesizing enzymes. We determined the synthesis of LTB4 and LTC4 by specific radioimmunoassay, and the messenger RNA (mRNA) expression of LT-synthesizing enzymes by reverse transcriptase polymerase chain reaction in peripheral polymorphonuclear leukocytes, which were obtained from controls and asthmatic children. The synthesis of LTB4 and LTC4, and the mRNA expression of 5 lipoxygenase, LTA4 hydrolase, and LTC4 synthase were enhanced in the patients. The mRNA expression of LT-synthesizing enzymes was up-regulated, resulting in increased LT synthesis, which may play an important role in the pathogenesis of childhood asthma. PMID- 12765317 TI - Analysis of sputum cell counts during spontaneous moderate exacerbations of asthma in comparison to the stable phase. AB - BACKGROUND: Acute airway inflammation is considered to characterize asthma exacerbations, but its specific cellular pattern has not yet been completely evaluated. AIM: To evaluate the prevalence of sputum eosinophilia during acute asthma exacerbations of moderate severity, compared with a stable phase of the disease, and to assess the concordance between changes in pulmonary function and sputum eosinophilia in the period between exacerbation and post exacerbation. METHODS: We compared sputum and blood inflammatory cell counts in 29 asthmatic subjects during a spontaneous moderate exacerbation of asthma (visit 1) with sputum and blood cell counts measured 4 weeks after the resolution of asthma exacerbation (visit 2). At visit 1, all subjects required an appropriate 1 week treatment with oral corticosteroids. RESULTS: At visit 1, all subjects were able to collect spontaneous sputum, whereas at visit 2 sputum was induced by inhalation of hypertonic saline (NaCl 3, 4, and 5%, 10 minutes each) with beta2 agonist pretreatment. Asthma exacerbation was accompanied by a significant increase in sputum eosinophil percentages compared with levels after exacerbation [25% (1-78) versus 4% (0-23), p<0.05). Only four subjects showed low sputum eosinophil percentages during exacerbation, and these showed no differences in main clinical findings with respect to subjects with sputum eosinophilia. At visit 2, the stability of asthma was assessed on the basis of PEF, FEV1, symptoms, and use of rescue beta2-agonist. Asthma was defined as stable in 21 out of 29 subjects. Sputum eosinophil percentages fell significantly between visit 1 and visit 2 in both stable and unstable patients, but at visit 2 sputum eosinophil percentages were still high in subjects with unstable asthma. In patients who proved to be stable at visit 2, there was a significant correlation between the changes recorded in sputum eosinophil percentages and in FEV1 between the two visits (rho: 0.723, p<0.001). CONCLUSION: Sputum cosinophil but not neutrophil percentages increase in most asthmatic subjects during moderate exacerbation of asthma. Changes in the degree of airway eosinophilic inflammation are related to changes in the severity of airway obstruction during asthma exacerbation. PMID- 12765318 TI - Hospital attendance prediction tool also identifies impaired quality of life in adults with asthma in general practice. AB - STUDY OBJECTIVE: To assess the ability of an adult risk screening questionnaire (RSQ), previously shown to predict attendance at hospital emergency departments, to identify impaired quality of life in adult patients with asthma in general practice. DESIGN: Baseline data from an RCT of asthma clinics in general practice, using the St. George Respiratory Questionnaire (SGRQ) to measure quality of life. SETTING: Twelve general practices in Adelaide, Australia. RESULTS: A total of 184 adult asthmatics were recruited, with a mean (standard deviation) age of 50.3 years (16.6 years). Age, gender, weight, number of comorbidities, smoking status, FEV1 and percent predicted FEV1 (pre-and post bronchodilator), peak flow (pre and post), and RSQ were independently related to SGRQ scores. After adjusting for potential confounders, an RSQ score predictive of hospital attendance was also associated with an average increase (worsening) in SGRQ total score of 11.9 (95% CI: 7.6, 16.3), an average increase in activity score of 9.0 (2.5, 15.5), symptom score of 14.8 (8.6, 21.0), and impact score of 13.1 (8.6, 17.6). These represent clinically significant differences. CONCLUSION: An RSQ score predictive of hospital attendance should alert the general practitioner to explore and address symptom, activity, and impact domains associated with impaired quality of life in adult asthma. PMID- 12765319 TI - The effects of asthma on pregnancy and perinatal outcomes. AB - The objective of the study was to examine the perinatal outcomes of women who have asthma and to determine if peak flow values change during pregnancy. A retrospective chart audit was conducted analyzing records of 567 women with asthma who delivered between 1992 and 1997. The most common maternal complications were meconium-stained amniotic fluid, preterm labor or delivery this pregnancy, oligohydramnios, and pregnancy-induced hypertension. The most common neonatal complications were meconium staining, preterm infant, and intrauterine growth restriction. Women who required systemic steroids were more likely to have oligohydramnios, intrauterine growth restricted infant, meconium staining, or pregnancy-induced hypertension. Peak flow values did not change by trimester of pregnancy. While the percentage of women who smoked was higher than the general population, smoking was not correlated with increased adverse perinatal outcomes. This study suggests an increased incidence of oligohydramnios, intrauterine growth restriction, and meconium-stained amniotic fluid in women with asthma. More research is needed to further the understanding of the relationship between asthma and perinatal outcomes. Prior studies have shown an association between poor control and adverse outcomes. Education is a major nursing implication. PMID- 12765320 TI - Pregnancy and birth outcomes in families with asthma. AB - Studies of maternal asthma in pregnancy have shown an increased risk of adverse neonatal and maternal outcomes such as preeclampsia, hypertension, cesarean delivery, prematurity, low birth weight, and perinatal/neonatal mortality. However, results are not consistent between studies. We studied the association between maternal asthma and various adverse neonatal and maternal outcomes and explored whether there is any evidence that pregnancy exacerbates maternal asthma. The data were collected as part of the Childhood Asthma Prevention Study. Pregnant women with asthma or women whose partners or other children had current symptoms of asthma were recruited at six Sydney hospitals. All women recruited were post 36 weeks gestation and were living within 30 km of the study recruitment center. Information about family history of asthma was collected using a questionnaire at 36 weeks gestation and subsequent information about antenatal and perinatal events was obtained from hospital records. Data from 611 pregnant women were available for analysis, 340 of whom had asthma. Hypertension was significantly more common in asthmatics than in nonasthmatics [OR = 2.16 (1.02-4.6), p < 0.043]. The prevalence of gestational diabetes, labor complications, delivery complications, and adverse neonatal outcomes did not differ significantly between the groups. We also found that the course of maternal asthma usually remains unchanged during pregnancy, but that more severe asthma is likely to get worse. We have confirmed previous observations that women with asthma are at increased risk of hypertension in pregnancy, which is consistent with studies that show that pregnant asthmatic women have a slightly increased risk of preeclampsia. However, we did not find evidence of an increased risk of adverse perinatal outcomes. PMID- 12765321 TI - Gender differences in health-related quality of life among asthma patients. AB - OBJECTIVE: To identify and explain differences between men and women with asthma regarding health-related quality of life (HRQoL). METHODS: A cross-sectional study was performed among 967 asthma patients recruited from general practice. Data were collected by means of a pulmonary function assessment, a face-to-face interview, and a written questionnaire. RESULTS: Women with asthma reported lower scores on HRQoL in the age groups 16-34 and 56-75 years but not in the age group 35-55 years. In all age groups, women reported more severe dyspnea but had higher levels of pulmonary function. The poorer HRQoL reported by women could be explained by a more severe dyspnea and a higher level of medication use in women. CONCLUSIONS: The finding that women with asthma aged 16-34 and 56-75 years report poorer HRQoL than men is not due to a more severe disease state in terms of pulmonary obstruction but does seem to be related to a more severe subjective disease state in women than in men. PMID- 12765322 TI - The Th1-Th2 paradigm and asthma: how far should we go? PMID- 12765323 TI - Perceptions and attitudes of adolescents with asthma. AB - BACKGROUND: Psychosocial factors can limit one's ability to effectively manage asthma. This can result in asthma morbidity that limits quality of life. While self-perceptions of asthmatic children and parents have been studied, less is known about self-perceptions of inner-city adolescents with asthma. OBJECTIVE: To examine perceptions and attitudes to treatment among inner-city adolescents with asthma. DESIGN/METHODS: We conducted a multistaged stratified sample survey at a high school located in the Bronx, NY. First, an asthma-screening survey was administered to 3800 registered students. Then, we identified a subset of 200 children with the most positive screening results for asthma. Next, a 32-item self-completion questionnaire was administered to this cohort. Students were surveyed regarding severity of asthma, perceptions and attitudes, and demographic information. Differences in proportions were tested by Chi-square analyses. Pearson rank-order correlation and multiple logistic regression were used to assess the relationship between perceptions and attitudes to self-treatment. RESULTS: A total of 160 (80%) students participated. Mean age was 15.7 years. Of the respondents, 63% were female. 68% were Hispanic, 26% were African American. 33% had weekly symptoms, and 14% had daily symptoms. Additionally, 41% did not know the name of their asthma medicine. Only 38% reported bringing an "asthma pump" when leaving the house. While 70% reported feeling in control over their asthma symptoms, 63% reported feelings of anxiety and 39% could remember a time when they felt like they were going to die from asthma. Subjects who reported feeling in control over their asthma were more likely to take an asthma pump with them when leaving the house (r = 72, p = 0.004). Only 39% had disclosed their asthma to their friends, and 29% felt embarrassed about having an asthma attack in front of their friends. Subjects who felt embarrassed about their asthma were significantly less likely to take their asthma pumps with them (r = 0.98, p < 0.001) and less likely to use their medications in front of their friends (r = .87, p < 0.001). CONCLUSIONS: Adolescents in this study reported feelings of anxiety, fear, and embarrassment about their asthma. Feelings of control over asthma symptoms were associated with positive attitudes toward self-treatment, while embarrassment was correlated with negative attitudes. These findings have implications for counseling adolescents with asthma about self-management strategies. PMID- 12765324 TI - A dosimetry study comparing NCS report-5, IAEA TRS-381, AAPM TG-51 and IAEA TRS 398 in three clinical electron beam energies. AB - New codes of practice for reference dosimetry in clinical high-energy photon and electron beams have been published recently, to replace the air kerma based codes of practice that have determined the dosimetry of these beams for the past twenty years. In the present work, we compared dosimetry based on the two most widespread absorbed dose based recommendations (AAPM TG-51 and IAEA TRS-398) with two air kerma based recommendations (NCS report-5 and IAEA TRS-381). Measurements were performed in three clinical electron beam energies using two NE2571-type cylindrical chambers, two Markus-type plane-parallel chambers and two NACP-02 type plane-parallel chambers. Dosimetry based on direct calibrations of all chambers in 60Co was investigated, as well as dosimetry based on cross calibrations of plane-parallel chambers against a cylindrical chamber in a high energy electron beam. Furthermore, 60Co perturbation factors for plane-parallel chambers were derived. It is shown that the use of 60Co calibration factors could result in deviations of more than 2% for plane-parallel chambers between the old and new codes of practice, whereas the use of cross-calibration factors, which is the first recommendation in the new codes, reduces the differences to less than 0.8% for all situations investigated here. The results thus show that neither the chamber-to-chamber variations, nor the obtained absolute dose values are significantly altered by changing from air kerma based dosimetry to absorbed dose based dosimetry when using calibration factors obtained from the Laboratory for Standard Dosimetry, Ghent, Belgium. The values of the 60Co perturbation factor for plane-parallel chambers (k(att) x k(m) for the air kerma based and p(wall) for the absorbed based codes of practice) that are obtained from comparing the results based on 60Co calibrations and cross-calibrations are within the experimental uncertainties in agreement with the results from other investigators. PMID- 12765325 TI - Random and systematic beam modulator errors in dynamic intensity modulated radiotherapy. AB - This paper reports on the dosimetric effects of random and systematic modulator errors in delivery of dynamic intensity modulated beams. A sliding-widow type delivery that utilizes a combination of multileaf collimators (MLCs) and backup diaphragms was examined. Gaussian functions with standard deviations ranging from 0.5 to 1.5 mm were used to simulate random positioning errors. A clinical example involving a clival meningioma was chosen with optic chiasm and brain stem as limiting critical structures in the vicinity of the tumour. Dose calculations for different modulator fluctuations were performed, and a quantitative analysis was carried out based on cumulative and differential dose volume histograms for the gross target volume and surrounding critical structures. The study indicated that random modulator errors have a strong tendency to reduce minimum target dose and homogeneity. Furthermore, it was shown that random perturbation of both MLCs and backup diaphragms in the order of sigma = 1 mm can lead to 5% errors in prescribed dose. In comparison, when MLCs or backup diaphragms alone was perturbed, the system was more robust and modulator errors of at least sigma = 1.5 mm were required to cause dose discrepancies greater than 5%. For systematic perturbation, even errors in the order of +/- 0.5 mm were shown to result in significant dosimetric deviations. PMID- 12765326 TI - Introducing a system for automated control of rotation axes, collimator and laser adjustment for a medical linear accelerator. AB - Mechanical stability and precise adjustment of rotation axes, collimator and room lasers are essential for the success of radiotherapy and particularly stereotactic radiosurgery with a linear accelerator. Quality assurance procedures, at present mainly based on visual tests and radiographic film evaluations, should desirably be little time consuming and highly accurate. We present a method based on segmentation and analysis of digital images acquired with an electronic portal imaging device (EPID) that meets these objectives. The method can be employed for routine quality assurance with a square field formed by the built-in collimator jaws as well as with a circular field using an external drill hole collimator. A number of tests, performed to evaluate accuracy and reproducibility of the algorithm, yielded very satisfying results. Studies performed over a period of 18 months prove the applicability of the inspected accelerator for stereotactic radiosurgery. PMID- 12765327 TI - Automatic feathering of split fields for step-and-shoot intensity modulated radiation therapy. AB - Due to leaf travel range limitations of the Varian Dynamic Multileaf Collimator (DMLC) system, an IMRT field width exceeding 14.5 cm is split into two or more adjacent abutting sub-fields. The abutting sub-fields are then delivered as separate treatment fields. The accuracy of the delivery is very sensitive to multileaf positioning accuracy. The uncertainties in leaf and carriage positions cause errors in the delivered dose (e.g., hot or cold spots) along the match line of abutting sub-fields. The dose errors are proportional to the penumbra slope at the edge of each sub-field. To alleviate this problem, we developed techniques that feather the split line of IMRT fields. Feathering of the split line was achieved by dividing IMRT fields into several sub-groups with different split line positions. A Varian 21EX accelerator with an 80-leaf DLMC was used for IMRT delivery. Cylindrical targets with varying widths (>14.5 cm) were created to study the split line positions. Seven coplanar 6 MV fields were selected for planning using the NOMOS-CORVUS system. The isocentre of the fields was positioned at the centre of the target volume. Verification was done in a 30 x 30 x 30 cm3 polystyrene phantom using film dosimetry. We investigated two techniques to move the split line from its original position or cause feathering of them: (1) varying the isocentre position along the target width and (2) introduction of a 'pseudo target' outside of the patient (phantom). The position of the 'pseudo target' was determined by analysing the divergence of IMRT fields. For target widths of 14-28 cm, IMRT fields were automatically split into two sub-fields, and the split line was positioned along the centre of the target by CORVUS. Measured dose distributions demonstrated that the dose to the critical structure was 10% higher than planned when the split line crossed through the centre of the target. Both methods of modifying the split line positions resulted in maximum shifts of approximately 1 cm from the original. Therefore, it was concluded that the feathering of the split line may be used for reducing the magnitude of hot/cold spots. This method was tested for an oesophageal cancer case. For a six-field arrangement, it was possible to create three field sub-groups with different split lines. The feathering technique developed in this work does not require any modifications of the radiation fields during the course of treatment because only one treatment plan is used to deliver the entire course of radiation treatments. In addition, this method may be more biologically effective because the split line feathering is achieved for every fraction of radiation. PMID- 12765328 TI - Plastic scintillation dosimetry: optimization of light collection efficiency. AB - Practical contemporary radiotherapy dosimetry systems used for dose measurement and verification are ionization chambers (which typically have at least a 0.1 cm3 air cavity volume), thermoluminescent dosimeters (TLDs) and silicon diodes. However, during the last decade, there has been an increased interest in scintillation dosimetry using small water-equivalent plastic scintillators, due to their favourable characteristics when compared with other more commonly used detector systems. Although plastic scintillators have been shown to have many desirable dosimetric properties, as yet there is no successful commercial detector system of this type available for routine clinical use in radiation oncology. The objectives of this study are to identify the factors preventing this new technology from realizing its full potential in commercial applications. A definition of signal to noise ratio (S/N) will be proposed for this category of detectors. In doing so the S/N ratio for an early prototype design has been calculated and/or measured. Criteria to optimize the response and sensitivity of this category of detectors are presented. PMID- 12765329 TI - Intraoperative dynamic dosimetry for prostate implants. AB - This paper describes analytic tools in support of a paradigm shift in brachytherapy treatment planning for prostate cancer--a shift from standard pre planning to intraoperative planning using dosimetric feedback based on the actual deposited seed positions within the prostate. The method proposed is guided by several desiderata: (a) bringing both planning and evaluation in the operating room (i.e. make post-implant evaluation superfluous) therefore making rectifications--if necessary--still achievable; (b) making planning and implant evaluation consistent by using the same imaging system (ultrasound); and (c) using only equipment commonly found in a hospital operating room. The intraoperative dosimetric evaluation is based on the fusion between ultrasound images and 3D seed coordinates reconstructed from fluoroscopic projections. Automatic seed detection and registration of the fluoroscopic and ultrasound information, two of the three key ingredients needed for the intraoperative dynamic dosimetry optimization (IDDO), are explained in detail. The third one, the reconstruction of 3D coordinates from projections, was reported in a previous article. The algorithms were validated using a custom-designed phantom with non radioactive (dummy) seeds. Also, fluoroscopic images were taken at the conclusion of an actual permanent prostate implant and compared with data on the same patient obtained from radiographic-based post-implant evaluation. To offset the effect of organ motion the comparison was performed in terms of the proximity function of the two seed distributions. The agreement between the intra- and post operative seed distributions was excellent. PMID- 12765330 TI - Treatment time reduction for large thermal lesions by using a multiple 1D ultrasound phased array system. AB - To generate large thermal lesions in ultrasound thermal therapy, cooling intermissions are usually introduced during the treatment to prevent near-field heating, which leads to a long treatment time. A possible strategy to shorten the total treatment time is to eliminate the cooling intermissions. In this study, the two methods, power optimization and acoustic window enlargement, for reducing power accumulation in the near field are combined to investigate the feasibility of continuously heating a large target region (maximally 3.2 x 3.2 x 3.2 cm3). A multiple 1D ultrasound phased array system generates the foci to scan the target region. Simulations show that the target region can be successfully heated without cooling and no near-field heating occurs. Moreover, due to the fact that there is no cooling time during the heating sessions, the total treatment time is significantly reduced to only several minutes, compared to the existing several hours. PMID- 12765331 TI - Automatic localization of implanted seeds from post-implant CT images. AB - An automatic localization method of implanted seeds from a series of post-implant computed tomography (CT) images is described in this paper. Post-implant CT studies were obtained for patients who underwent prostate brachytherapy. Bright areas were segmented using binary thresholding in each CT slice, and geometrical information on these areas was collected. Large areas (possibly containing two connected seeds) were split into smaller ones by geometry-based filtering in each slice. The area connectivity along the longitudinal direction was analysed using a geometry-based connection search algorithm executed on every area slice by slice, so that the connected areas were combined into one object. The weighted centroid of each object was taken as the seed position. This method was tested on a seed-containing prostate phantom as well as using CT studies from patients. Statistical analysis demonstrates that it can achieve above 99% detection rate with reliable localization accuracy and high speed. It is reliable and convenient for localizing implanted seeds on CT and can be used to assist post-implant dosimetry for prostate brachytherapy. PMID- 12765332 TI - Effect of dextran-induced changes in refractive index and aggregation on optical properties of whole blood. AB - The purpose of the present study is to investigate systematically the mechanisms of alterations in the optical properties of whole blood immersed in the biocompatible agent dextran, and to define the optimal concentration of dextrans required for blood optical clearing in order to enhance the capability of light penetration depth for optical imaging applications. In the experiments, dextrans with different molecular weights and various concentrations were employed and investigated by the use of the optical coherence tomography technique. Changes in light attenuation, refractive index and aggregation properties of blood immersed in dextrans were studied. It was concluded from the results that the mechanisms for blood optical clearing are characteristic of the types of dextrans employed, their concentrations and the application stages. Among the substances applied, Dx500 at a concentration at 0.5 g dl(-1) gives the best result in improving light penetration depth through the blood. The increase of light transmission at the beginning of the addition of dextrans is mainly attributed to refractive index matching between the scattering centres and the ground matter. Thereafter, the transmission change is probably due to a dextran-induced aggregation disaggregation effect. Overall, light scattering in the blood could be effectively reduced by the application of dextrans. It represents a promising approach to increasing the imaging depth for in vivo optical imaging of biological tissue, for example optical coherence tomography. PMID- 12765333 TI - Intensity-modulated radiation therapy using a variable-aperture collimator. AB - This paper extends some earlier concepts of using a tertiary mask plus jaws for delivering IMRT without a multileaf collimator. The new concept is to sweep a variable-aperture collimator (VAC) across the space of the intensity-modulated beam (IMB) to be delivered and to strip this IMB down into multiple-static-field components, each deliverable with the VAC. The stripping algorithm is described and it is shown, for several designs of VAC, that the mean number of field components and mean number of monitor units is less using the VAC than would be required for a jaws-only (JO) decomposition. The VAC would be simpler to construct than several previously suggested jaws-plus-mask (J+M) combinations. As well as describing a simple VAC for the use with jaws, we propose a design concept of a hybrid VAC. We also show that adding the potential to rotate the simple or hybrid VAC for some components relative to the field to be modulated is advantageous. PMID- 12765334 TI - Schizophrenia: neural mechanisms for novel therapies. AB - Although valuable antischizophrenic drugs exist, they only partially ameliorate symptoms and elicit substantial side effects. Classic neuroleptic drugs act by blocking dopamine receptors. They can relieve some symptoms but not behavioral withdrawal features that are designated "negative" symptoms. Clozapine and related newer atypical neuroleptics may be more efficacious in relieving negative symptoms. Understandng their actions may facilitate new drug discovery. Agents influencing glutamate neurotransmission and N-methyl-D-aspartate receptors, especially the cotransmitter D-serine, are promising. Stimulation of the alpha7 subtype of nicotinic acetylcholine receptor may also be efficacious. The search for genes linked to schizophrenia has revealed several leads that may permit development of novel therapeutic agents. Promising genes include disrupted-in schizophrenia-1, dysbindin, and neuregulin. PMID- 12765335 TI - Interleukin-10 deficiency increases atherosclerosis, thrombosis, and low-density lipoproteins in apolipoprotein E knockout mice. AB - Interleukin (IL)-10 is an anti-inflammatory cytokine that may play a protective role in atherosclerosis. The aim of this study was to assess the effect of IL-10 deficiency in the apolipoprotein E knockout mouse. Apolipoprotein E deficient (E /-) and IL-10 deficient (-/-) mice were crossed to generate E-/- x IL-10-/- double knockout mice. By 16 wk, cholesterol and triglycerides were similar in double and single knockouts but the lack of IL-10 led to increased low-density lipoprotein cholesterol whereas very-low-density lipoprotein was reduced. In parallel, T-helper 1 responses and lesion size were dramatically increased in double knockout compared with E-/- controls. At 48 wk, matrix metalloproteinases and tissue factor activities were increased in lesions of double-knockout mice. Furthermore, markers of systemic coagulation were increased, and vascular thrombosis in response to i.v. thrombin occurred more frequently in E-/- x IL-10 /- than in E-/- mice. Our findings suggest that IL-10 deficiency plays a deleterious role in atherosclerosis. The early phase of lesion development was increased, and the proteolytic and procoagulant activity was elevated in advanced lesions. These data show that IL-10 may reduce atherogenesis and improve the stability of plaques. PMID- 12765336 TI - Cloning and characterization of Cbl-associated protein splicing isoforms. AB - Cbl-associated protein (CAP) is an adaptor protein that plays important roles in both signal transduction and cytoskeleton rearrangement. Alternative splicing of the gene SORBS1 results in multiple isoforms of CAP protein. We report here the cloning of 3 new CAP isoforms, CAP2, CAP3, and CAP4, from mouse adipose tissue. RT-PCR analyses reveal that the isoform mRNAs are differentially expressed. CAP2, CAP3, and CAP4 contain a coiled-coil domain. In addition, CAP4 contains a proline rich region, part of which exists in CAP3. Coimmunoprecipitation experiments show that CAP4 forms a homodimeric complex. While these new isoforms similarly interact with Cbl, they exhibit varied binding specificity toward vinculin. In contrast to CAP1 and CAP2, CAP4 does not interact with vinculin, and CAP3 binds with low affinity. Immunofluorescence analysis demonstrates differential subcellular localization of Myc-tagged CAP isoforms in 3T3-L1 adipocytes. These results suggest that these new isoforms of CAP might play different signaling roles. PMID- 12765339 TI - Characterization of dominant-negative forms of anthrax protective antigen. AB - Certain mutations within the protective antigen (PA) moiety of anthrax toxin endow the protein with a dominant-negative (DN) phenotype, converting it into a potent antitoxin. Proteolytically activated PA oligomerizes to form ring-shaped heptameric complexes that insert into the membrane of an acidic intracellular compartment and promote translocation of bound edema factor and/or lethal factor to the cytosol. DN forms of PA co-oligomerize with the wild-type protein and block the translocation process. We prepared and characterized 4 DN forms: a single, a double, a triple, and a quadruple mutant. The mutants were made by site directed mutation of the cloned form of PA in Escherichia coli and tested by various assays conducted on CHO cells or in solution. All 4 mutant PAs were competent for heptamerization and ligand binding but were defective in the pH dependent functions: pore formation, ability to convert to the SDS-resistant heptamer, and ability to translocate bound ligand. The single mutant (F427K) showed less attenuation than the others in the pH-dependent functions and lower DN activity in a CHO cell assay. The quadruple (K397D + D425K + F427A + 2beta2 2beta3) deletion showed the most potent DN activity at low concentrations but also gave indications of low stability in a urea-mediated unfolding assay. The double mutant (K397D + D425K) and the triple (K397D + D425K + F427A) showed strong DN activity and slight reduction in stability relative to the wild-type protein. The properties of the double and the triple mutants make these forms worthy of testing in vivo as a new type of antitoxic agent for treatment of anthrax. PMID- 12765337 TI - Dihydrobetulinic acid induces apoptosis in Leishmania donovani by targeting DNA topoisomerase I and II: implications in antileishmanial therapy. AB - Leishmaniasis is the second-most dreaded parasitic disease in the modern world, behind malaria. The lack of effective vaccines demand improved chemotherapy along with the development of lead compounds and newer targets. We report here that the pentacyclic triterpenoid, dihydrobetulinic acid (DHBA), is a novel lead compound for antileishmanial therapy. It acts by targeting DNA topoisomerases. DNA topoisomerase I and II activity was studied using relaxation and decatenation assays. Mechanistic studies were based on the decreased mobility of enzyme-bound DNA compared with free DNA and the differential mobility of nicked and supercoiled monomers in 1% agarose gel. Pulsed field gradient gel electrophoresis, confocal microscopy, and transmission electron microscopy were performed to assess cytotoxicity of the compound and ultrastructural damage of the parasite. Apoptosis was studied by the isolation of DNA from DHBA-treated parasites and subsequent electrophoresis in 1% agarose gel. DHBA inhibits growth of Leishmania donovani promastigotes and amastigotes with an IC50 of 2.6 and 4.1 microM respectively. The compound is a dual inhibitor of DNA topoisomerases that fails to induce DNA cleavage and acts by preventing the formation of enzyme-DNA binary complex, ultimately inducing apoptosis. Treatment of infected golden hamsters with the compound markedly reduces (> 92%) parasitic burden, both in spleen and liver. Interestingly, the 17-decarboxylated analogue, dihydrolupeol, does not inhibit DNA topoisomerase I and II, has no effect on parasitic growth, and also fails to induce apoptosis. DHBA is a potent antileishmanial agent that induces apoptosis by primarily targeting DNA topoisomerases. Therefore it is a strong candidate for use in designing new antileishmanial drugs. PMID- 12765338 TI - Structural basis for the proinflammatory cytokine activity of high mobility group box 1. AB - High mobility group box 1 (HMGB), a ubiquitous DNA-binding protein, has been implicated as a proinflammatory cytokine and late mediator of lethal endotoxemia. HMGB1 is released by activated macrophages. It amplifies and extends the inflammatory response by inducing cytokine release and mediating acute lung injury, anorexia, and the inflammatory response to tissue necrosis. The kinetics of HMGB1 release provide a wide therapeutic window for endotoxemia because extracellular levels of HMGB1 begin to increase 12 to 24 h after exposure to inflammatory stimuli. Here, we demonstrate that a DNA-binding domain of HMGB1, the B box, recapitulates the cytokine activity of full length HMGB1 and efficiently activates macrophages to release tumor necrosis factor (TNF) and other proinflammatory cytokines. Truncation of the B box revealed that the TNF stimulating activity localizes to 20 amino acids (HMGB1 amino acids 89 to 108). Passive immunization of mice with antibodies raised against B box conferred significant protection against lethal endotoxemia or sepsis, induced by cecal perforation. These results indicate that a proinflammatory domain of HMGB1 maps to the highly conserved DNA-binding B box, making this primary sequence a suitable target in the design of therapeutics. PMID- 12765340 TI - A plasminogen activator inhibitor-1 promoter polymorphism and idiopathic interstitial pneumonia. AB - The normal fibrinolytic activity within the alveolar space is suppressed in fibrotic lung diseases in part because of increased levels of plasminogen activator inhibitor-1 (PAI-1). Studies with animals have shown that inhibition of the plasminogen system by PAI-1 increases the generation of pulmonary fibrosis. To determine if a similar relationship occurs in human fibrotic lung diseases, we took advantage of a polymorphism (4G/5G) that occurs in the promoter region of the human PAI-1 gene and influences the expression of PAI-1. We hypothesized that the 4G/4G genotype, because of its association with higher levels of PAI-1, would occur in patients with idiopathic interstitial pneumonia more frequently than in a control population. PAI-1 promoter genotype was determined in 88 well characterized patients with idiopathic interstitial pneumonia consisting of 62 patients with usual interstitial pneumonia and 26 with nonspecific interstitial pneumonia. DNA was extracted from paraffin-embedded biopsy tissue and the genotype identified by polymerase chain reaction and restriction endonuclease digestion. We found that the distribution of PAI-1 genotypes in the idiopathic interstitial pneumonia population was similar to that of a large control population. However, subgroup analysis showed that patients with nonspecific interstitial pneumonia were more likely than the control population to have the promoter genotype (4G/4G) that is associated with higher levels of PAI-1. A similar pattern in PAI-1 polymorphism was not seen in the usual interstitial pneumonia subgroup. The results of this study support the conclusion that PAl-1 expression influences the development of nonspecific interstitial pneumonia in a similar manner to what occurs in animal models of pulmonary fibrosis. Patients with usual interstitial pneumonia did not show the same relationship with PAl-1 genotype. PMID- 12765342 TI - The North Shore-Long Island Jewish Research Institute. AB - The North Shore-Long Island Jewish Research Institute is directed by Nicholas Chiorazzi, MD. It provides investigators from a variety of scientific fields and backgrounds with the opportunity to focus on disease-oriented medical research. The goal of the institute is to understand the underlying biological processes of disease and ultimately to develop new and more effective therapies for patients. The affiliation with the North Shore-Long Island Jewish Health System and its patients provides a unique opportunity for focused biomedical and translational research. PMID- 12765341 TI - Hypertension and angiotensin II hypersensitivity in aminopeptidase A-deficient mice. AB - Local concentrations of the vasopressor peptide, angiotensin II (AngII), depend upon the balance between synthesis and degradation. Previous studies of blood pressure (BP) regulation have focused primarily on the generation of AngII and its receptors, and less attention has been devoted to angiotensin degradation. Aminopeptidase A (APA, EC 3.4.11.7) is responsible for the N-terminal cleavage of AngII, a hydrolytic event that serves as a rate-limiting step in angiotensin degradation. To evaluate the physiological role of APA, we examined BP homeostasis in APA-deficient mice. We measured basal BP and BP with continuous infusion of AngII in APA mutant mice by tail-cuff method. We also evaluated the development and histology of AngII-targeted organs as well as urine excretion in these mice. Homozygous APA mutant mice were found to have elevated basal systolic BP when compared with heterozygous mutant and wild-type littermate mice. Infusion of AngII led to an enhanced systolic BP response in the APA-deficient mice. Despite the sustained elevation of BP in APA knockout mice, neither their renal and cardiac sizes nor their histological appearances were not different from control mice. Moreover, the volume, osmolality, and electrolyte content of the urine were normal in APA-deficient mice. APA deficiency increased baseline BP and enhanced the hypertensive response to increased levels of AngII. These findings indicate a physiological role for APA in lowering BP and offer novel insight into the mechanisms for developing hypertension. PMID- 12765343 TI - Pre-ovulatory granulosa cells of infertile women with endometriosis are less sensitive to luteinizing hormone. AB - PROBLEM: Reduced fertilization rates in women with minor endometriosis may be the result of direct effects on the ovary or to primary dysfunction within the hypothalamic-pituitary-ovarian axis. This controlled study was designed to examine the steroidogenic potential of luteinized granulosa cells in women with minor endometriosis. METHOD OF STUDY: Granulosa cells were harvested at oocyte recovery and incubated for 3 hr in increasing concentrations of luteinizing hormone (LH). The dissociation constant for added concentrations of LH was computed (as Km LH) and the results were compared between women with endometriosis and controls. RESULTS: Women with minor endometriosis had a higher dissociation constant than women with tubal damage [Km 0.98 (0.58-9.24) versus 0.33 (0.28-0.72) ng/mL, P = 0.019], indicating reduced sensitivity to LH. CONCLUSIONS: In women with endometriosis, granulosa cells were less sensitive to LH stimulation. This provides further evidence for primary ovarian dysfunction as a significant contributory cause of the associated subfertility. PMID- 12765344 TI - Increased pregnancy-associated plasma protein-A (PAPP-A) concentrations in peritoneal fluid of women with endometriosis. AB - PROBLEM: Pregnancy-associated plasma protein-A (PAPP-A) belongs to a group of glycoproteins isolated from extracts of human placenta. Healthy ovarian and uterine tissues are also known to express PAPP-A. We hypothesized that PAPP-A levels might also be elevated in the peritoneal fluid (PF) of women with endometriosis, and examined variations in PF PAPP-A during the menstrual cycle and with the severity of the disease. METHOD OF STUDY: PF PAPP-A were measured in 60 women with endometriosis and 38 women without endometriosis using a high sensitivity immunofluorometric assay. RESULTS: We found that the mean level of PAPP-A was higher in PF from patients with endometriosis than controls (p = 0.003). Furthermore, significant correlation was found between the stages of endometriosis and the levels of PAPP-A in these patients (r = 0.39, p = 0.009). The concentrations of PAPP-A in PF were significantly higher in the secretory phase than the proliferative phase of the menstrual cycle in both women with and without endometriosis (p = 0.009 and P = 0.002, respectively). Finally, among the controls, women undergoing tubal ligation had significantly lower mean PF levels of PAPP-A than women with infertility or pelvic pain (p = 0.001). CONCLUSION: We conclude that PF levels of PAPP-A vary during the menstrual cycle, and are highest in the secretory phase. We also find that PF PAPP-A levels are significantly increased in women with endometriosis, and that the degree of elevation corresponds to the extent of disease. PMID- 12765345 TI - Differential expression of interleukins (IL)-13 and IL-15 in ectopic and eutopic endometrium of women with endometriosis and normal fertile women. AB - PROBLEM: Interleukins (IL) 13 and 15 are key regulators of inflammatory and immune responses, processes that are central to endometriosis and associated abnormalities. The present study examined (1) whether ectopic endometrial tissue expresses IL-13 and IL-15 (2) if their expression differs compared with matched eutopic endometrium and control endometrium from normal fertile women, and (3) if peritoneal fluids (PF) content of these cytokines reflects the disease compared with PF from women with peritoneal adhesions unrelated to endometriosis and those without pelvic pathology. METHODS: The expression of IL-13 and IL-15 mRNA and protein was determined using quantitative RT-PCR, ELISA and immunohistochemistry. RESULTS: Ectopic endometrium expresses IL-13 and IL-15 mRNA and protein with elevated levels compared with eutopic and control endometrium, irrespective of the phases of the menstrual cycle, with predominance in IL-13 expression. Endometrial epithelial cells were found to be the primary site of IL-13 and IL-15 expression. The PF content of IL-13 and IL-15 show a trend toward higher concentrations in women with adhesion and endometriosis, respectively, compared with fertile control without pelvic pathology. CONCLUSION: Interleukins 13 and 15 are expressed in ectopic endometrium and present in PF of women with endometriosis and their elevated expression in ectopic endometrium suggests that these cytokines play a key role in local inflammatory/immune responses that are critical in endometriosis-associated abnormalities. PMID- 12765346 TI - Intravenous immunoglobulin treatment in women with recurrent abortions: increased cytokine levels and reduced Th1/Th2 lymphocyte ratio in peripheral blood. AB - PROBLEM: The aim of this study was to investigate changes in peripheral blood Th1/Th2 cytokine levels and lymphocyte ratios after massive intravenous immunoglobulin (MIVIg) treatment for women with recurrent spontaneous abortion (RSA) of unexplained etiology. METHOD OF STUDY: Serum Th1 (IFN-gamma, TNF-alpha) and Th2 cytokine (IL-4, IL-10) levels were assessed by ELISA methods (n = 9) and peripheral blood Th1/Th2 lymphocyte ratios (n = 4) by flow cytometry before and after MIVIg treatments in women with four or more consecutive RSA. RESULTS: Pre treatment serum IFN-gamma (0.06 +/- 0.09 pg/mL, mean +/- SD), TNF-alpha (0.21 +/- 0.45 pg/mL), IL-4 (0.70 +/- 1.16 pg/mL), and IL-10 (1.12 +/- 1.67 pg/mL) increased to 0.17 +/- 0.16 pg/mL, 0.77 +/- 0.28 pg/mL, 1.82 +/- 0.89 pg/mL, and 3.44 +/- 0.48 pg/mL, respectively, after MIVIg treatments (P < 0.05). CD4 positive IFN-gamma/IL-4 lymphocyte ratios (17.3 +/- 9.1) were reduced to 11.5 +/- 7.1 after treatment (P < 0.05). CONCLUSIONS: Massive intravenous immunoglobulin treatments increased peripheral blood cytokine levels and decreased Th1/Th2 lymphocyte ratios; thus, MIVIg treatments modify the peripheral Th1/Th2 balance. PMID- 12765347 TI - Successful pregnancy with low molecular weight heparin in two women with recurrent miscarriage of unknown etiology. AB - We report here two cases of recurrent miscarriages that were successfully treated with continuous intravenous administration of low molecular weight heparin (LMWH). One patient experienced 11 spontaneous abortions, and the other eight abortions. Previous treatments including prednisone, aspirin and mononuclear-cell immunization were all unsuccessful. They were negative for anticardiolipin antibodies and lupus anticoagulant, and had no inherited thrombophilic disorder. Intravenous administration of LMWH, 4800 units of dalteparin, was started as soon as the conception was confirmed, and was continued until 34 weeks of gestation. They were delivered of live born infants. PMID- 12765348 TI - Macrophages in human reproductive tissues contain luteinizing hormone/chorionic gonadotropin receptors. AB - PROBLEM: To determine whether macrophages in human reproductive tissues contain luteinizing hormone (LH)/human chorionic gonadotropin (hCG) receptor mRNA and receptor protein that can bind 125I-hCG. METHOD OF STUDY: Macrophages isolated from term pregnancy human decidua were used for LH/hCG receptor detection by in situ hybridization for receptor mRNA and immunocytochemistry for a macrophage marker, CD68, performed alone and in combination, reverse transcription-nested polymerase chain reaction, Western and ligand blotting. The LH/hCG receptor presence in macrophages in late luteal phase human endometria and corpora lutea was determined by sequential performance of in situ hybridization and immunocytochemistry. RESULTS: The macrophages present in term pregnancy human decidua and late luteal phase human endometria and corpora lutea contain LH/hCG receptors. CONCLUSIONS: This is the first demonstration of macrophages present in human reproductive tissues containing LH/hCG receptors. The receptor presence suggests that LH and hCG may regulate macrophage functions in gonadal as well as in non-gonadal target tissues. PMID- 12765349 TI - Demonstration of the presence of IL-16, IL-17 and IL-18 at the murine fetomaternal interface during murine pregnancy. AB - PROBLEM: To determine if interleukin-16 (IL-16), IL-17, and IL-18 are present at the murine fetomaternal interface during pregnancy as a first step towards investigating their roles in fetomaternal relationship. METHODS: Expression of IL 16, IL-17, and IL-18, was assessed by immunohistochemistry (IHC) in the BALB/c x BALB/k (H2d x H2k), and the CBA/J x BALB/c non-abortion prone, and CBA/J x DBA/2 abortion prone matings. Enzyme-linked immunosorbent assay (ELISA) were performed for the two latter cytokines to compare local production in the abortion prone CBA/J x DBA/2 versus the non-abortion prone CBA/J x BALB/c matings. RESULTS: Expression of IL-17 was borderline. The anti-IL-16 staining specifically localized in the uterine stroma and glandular epithelium and was rather low in the placenta. IL-18 staining started in the peri-implantation uterus in the basal proliferative stroma, and was also traced, although weaker, in the glandular epithelium. In the immediate post-implantation period, a weak stromal staining persisted but there was a strong labeling of the ectoplacental cone. Interestingly, when the ectoplacental cone differentiates into placenta having a major histocompatibility complex (MHC) class I + spongiotrophoblast and a (MHC class I-) labyrinth, a very strong transient labeling of uterine natural killer (u-NK) cells was found. Later in gestation, IL-18 was also produced by giant cell and spongiotrophoblast. Finally, we compared by ELISA the production of IL-17/-18 in CBA/J x DBA/2 and CBA/J x BALB/c matings. We detected significantly more IL-18 in the non-abortion prone combination decidua or placenta. CONCLUSION: The three cytokines IL-16, IL-17, and IL-18 were detected at the fetomaternal interface with a tissue specific, stage-dependent distribution. The predominance of IL-18 secretion in the non-resorption prone matings lead us to question the general validity of the classical T-helper (Th)1/2 paradigm. PMID- 12765350 TI - Insulin-like growth factor II in gynecological cancers: a preliminary study. AB - PROBLEM: We have previously reported elevated serum levels of cervical human papilloma viral proteins E6 and E7 and serum insulin-like growth factor II (IGF II) in women with cervical cancer and advanced cervical intraepithelial neoplasia. As most women with cervical cancer have elevated levels of serum IGF II, we sought to determine whether the cervical cancer and lymph node biopsies from these women demonstrated increased production of IGF-II and whether this elevation was also present in ovarian and endometrial cancers. METHOD OF STUDY: We used the semi-quantitative immunofluorescent antibody assay established in our laboratory to identify the levels of IGF-II in 21 cervical cancers (seven with matching lymph nodes), 18 benign cervical biopsies, 13 endometrial cancers, 15 benign endometrial biopsies, 5 ovarian cancers, and 15 benign ovarian biopsies. RESULTS: The immunofluorescent IGF-II levels (relative intensity per pixel) were the highest in cervical cancers; they were significantly higher than in matched controls. IGF-II levels were not higher in ovarian cancers and only slightly elevated in endometrial cancers. The presence of IGF-II in pelvic lymph nodes of women with cervical cancer paralleled with those in the cervical cancers. Interestingly, we could identify small nests of metastases of malignant cells in the nodes (pauci-cellular metastasis) by using IGF-II as the marker. CONCLUSION: We propose that measurement and identification of IGF-II in the cervical biopsy may be a sensitive method of detecting cervical cancer and metastatic spread in the lymph nodes. PMID- 12765351 TI - Collagenase-3 (MMP-13) in fetal membranes and amniotic fluid during pregnancy. AB - OBJECTIVE: To examine the expression, site of production and a role of collagenase-3 in human fetal membranes and amniotic fluid (AF). METHODS: Amniochorion collected at the time of elective repeat cesareans prior to labor from normal term gestation, were placed in an organ explant system for 72 hr. The AF was collected from the following groups of women: women at term; women at third trimester pregnancy; women at mid-trimester pregnancy. Women with premature rupture of the membranes (PROM) with or without pre-term labor and infection delivered by cesareans before term; women with pre-term labor and no rupture of membranes and delivered by cesareans before term. The mRNA expression of collagenase-3 was studied by reverse transcriptase-polymerase chain reaction (RT PCR) and protein concentrations in the AF were assayed by enzyme-linked immunosorbent assay (ELISA). Site of collagenase-3 production was documented by immunohistochemistry. Statistical comparisons were made using ANOVA. All statistical hypotheses were adjusted for multiple comparisons using the Scheffe method of adjustment. RESULTS: Amniochorion at term expressed collagenase-3 mRNA. Immunohistochemistry localized collagenase-3 in both amnion and chorion. Collagenase-3 was seen in AF from all gestational age groups with lowest levels at mid-trimester [167.8 pg/mL] and highest during term labor (323.92 pg/mL) with no statistically significant difference between term and third trimester (310.11 pg/mL). AF levels of collagenase-3 were significantly higher in women with pre term labor (370.02 pg/mL; P(adj) = 0.046) or pre-term labor and infection (628.73 pg/mL; P(adj) = 0.002) compared with PROM with pre-term labor and infection (87.19 pg/mL). CONCLUSION: Collagenase-3 mRNA is constitutively expressed and produced in amniochorion. It is a normal physiologic constituent of AF. PROM is not associated with an increase in the AF collagenase-3. PMID- 12765352 TI - Polyhydroxyalkanoates: an overview. AB - Polyhydroxyalkanoates have gained major importance due to their structural diversity and close analogy to plastics. These are gaining more and more importance world over. Different sources (natural isolates, recombinant bacteria, plants) and other methods are being investigated to exert more control over the quality, quantity and economics of poly(3-hydroxybutyrate) (PHB) production. Their biodegradability makes them extremely desirable substitutes for synthetic plastics. The PHB biosynthetic genes phbA, phbB and phbC are clustered and organized in one phbCAB operon. The PHB pathway is highly divergent in the bacterial genera with regard to orientation and clustering of genes involved. Inspite of this the enzymes display a high degree of sequence conservation. But how similar are the mechanisms of regulation of these divergent operons is as yet unknown. Structural studies will further improve our understanding of the mechanism of action of these enzymes and aid us in improving and selecting better candidates for increased production. Metabolic engineering thereafter promises to bring a feasible solution for the production of "green plastic". PMID- 12765353 TI - Biomethanation under psychrophilic conditions: a review. AB - Anaerobic digestion of animal manure, sewage and other agricultural wastes at psychrophilic temperatures has not been explored as extensively as either mesophilic or thermophilic digestion, probably due to little anticipation of the development of economically attractive systems using this technology. This review article discusses psychrophilic anaerobic digestion studies reported by various researchers using different substrates. The effect of operational parameters such as type of substrate, size of inoculum, concentration of volatile fatty acids, hydraulic retention time and loading rate, on reduction of TS/VS, BOD/COD and biogas yield is discussed in detail. PMID- 12765354 TI - Bioeffects of microwave--a brief review. AB - Since the 18th century scientists have been intrigued by the interaction of electromagnetic fields (EMFs) and various life processes. Attention has been focussed on EMFs in different frequency ranges, of which microwave frequency range forms an important part. Microwaves are part of the electromagnetic spectrum and are considered to be that radiation ranging in frequency from 300 million cycles per second (300 MHz) to 300 billion cycles per second (300 GHz), which correspond to a wavelength range of 1 m down to 1 mm. This nonionising electromagnetic radiation is absorbed at molecular level and manifests as changes in vibrational energy of the molecules or heat (Microwaves irradiating the community, Hidden hazards, Bantan Books publisher, Australia, 1991). Identifying and evaluating the biological effects of microwaves have been complex and controversial. Because of the paucity of information on the mechanism of interaction between microwave and biological systems, there has been a persistent view in physical and engineering sciences, that microwave fields are incapable of inducing bioeffects other than by heating (Health Physics 61 (1991) 3). Of late, the nonthermal effects of microwaves on tissue responses are being documented (Physiol. Rev. 61 (1981) 435; Annals of New York Acad. Sci. 247 (1975) 232; J. Microwave Power 14 (1979) 351; Bioelectromagnetics 7 (1986a) 45; Bioelectromagnetics 7 (1986b) 315; Biologic Effects and Health Hazards of Microwave Radiation, Warsaw, Polish Medical Publication (1974) 289; Biologic Effects and Health hazards of the microwave Radiation, Warsaw, Polish Medical Publication (1974) 22; Multidisciplinory perspectives in event-related brain potential research, Washington DC, US Environmental Protection Agency, (1978) 444). The present article is an attempt to familiarise the reader with pertinent information regarding the effects, mainly athermal, of microwave irradiation on biologic systems, especially microorganisms. PMID- 12765355 TI - A review of survival of pathogenic bacteria in organic waste used in biogas plants. AB - Anaerobic digestion is one way of handling biowaste and generating energy in the form of methane (biogas). The digested residue may be used as fertiliser on agricultural land. Biowaste is known to contain pathogenic bacteria such as Salmonella and other microorganisms that may be a health risk for both people and animals. The biosecurity risk associated with using digested residue as fertiliser is hard to assess, but this risk cannot be neglected. It is of greatest importance that the treatment in the biogas plants (BGPs) minimise the survival of pathogens. Temperature is the most important factor when considering the reduction of pathogens in BGP, but there are also other factors involved. Different indicator bacteria are used to evaluate the hygienic treatment, but an indicator that is good enough to give an overall picture has not yet been found. PMID- 12765356 TI - Transformation of vegetable waste into value added products: (A) the upgrading concept; (B) practical implementations. AB - Waste can contain many reusable substances of high value. Depending on there being an adequate technology this residual matter can be converted into commercial products either as raw material for secondary processes, as operating supplies or as ingredients of new products. Numerous valuable substances in food production are suitable for separation and recycling at the end of their life cycle, even though present separation and recycling processes are not absolutely cost efficient. In Part A a need statement is visualised--based on a holistic concept of food production--for the vegetable industry, recording occurrence, quantities and utilisation of the residual products. A literature survey, covering more than 160 articles from all over the world, plus our own investigations summarises the latest knowledge in the above-mentioned field and outline prospects for future economic treatment of vegetable 'co-products'. The main goal of a clean production process is demonstrated by three practical implementations in Part B: 1. Upgrading of vegetable residues for the production of novel types of products: multifunctional food ingredients in fruit juice and bakery goods. 2. Bioconversion via solid-state fermentation: vegetable residues as an exclusive substrate for the generation of fruity food flavours. 3. Conversion of vegetable residues into operating supplies: bioadsorbents for waste water treatment. The investigations are promising with regard to future application in the mentioned industrial branch. The outlined concept can be naturally transferred to several areas of industrial food production. The intentions of this research area are located at the development of techniques, which fulfil the conditions of environmental protection with costs to a minimum. The prospect of several new niche markets is worthwhile indeed. PMID- 12765357 TI - Fluctuation theory and (very) early statistical energy analysis (SEA). PMID- 12765358 TI - Expression for the estimation of time-averaged acoustic energy density using the two-microphone method. PMID- 12765359 TI - Evoked potential recording during echolocation in a false killer whale Pseudorca crassidens. PMID- 12765360 TI - Theory and experiment of Fourier-Bessel field calculation and tuning of a pulsed wave annular array. AB - A one-dimensional (1D) Fourier-Bessel series method for computing and tuning (beamforming) the linear lossless field of flat pulsed wave annular arrays is developed and supported with both numerical simulation and experimental verification. The technique represents a new method for modeling and tuning the propagated field by linking the quantized surface pressure profile to a known set of limited diffraction Bessel beams propagating into the medium. This enables derivation of an analytic expression for the field at any point in space and time in terms of the transducer surface pressure profile. Tuning of the field then also follows by formulating a least-squares design for the transducer surface pressure with respect to a given desired field in space and time. Simulated and experimental results for both field computation and tuning are presented in the context of a 10-ring annular array operating at a central frequency of 2.5 MHz in water. PMID- 12765361 TI - Measuring the porosity and the tortuosity of porous materials via reflected waves at oblique incidence. AB - An ultrasonic reflectivity method is proposed for measuring porosity and tortuosity of porous materials having a rigid frame. Porosity is the relative fraction by volume of the air contained within a material. Tortuosity is a geometrical parameter which intervenes in the description of the inertial effects between the fluid filled the porous material and its structure at high frequency range. It is generally easy to evaluate the tortuosity from transmitted waves, this is not the case for porosity because of its weak sensitivity in transmitted mode. The proposed method is based on measurement of reflected wave by the first interface of a slab of rigid porous material. This method is obtained from a temporal model of the direct and inverse scattering problems for the propagation of transient ultrasonic waves in a homogeneous isotropic slab of porous material having a rigid frame [Z. E. A. Fellah, M. Fellah, W. Lauriks, and C. Depollier, J. Acoust. Soc. Am. 113, 61 (2003)]. Reflection and transmission scattering operators for a slab of porous material are derived from the responses of the medium to an incident acoustic pulse at oblique incidence. The porosity and tortuosity are determined simultaneously from the measurements of reflected waves at two oblique incidence angles. Experimental and numerical validation results of this method are presented. PMID- 12765362 TI - Finite difference computation of head-related transfer function for human hearing. AB - Modeling the head-related transfer function (HRTF) is a key to many applications in spatial audio. To understand and predict the effects of head geometry and the surrounding environment on the HRTF, a three-dimensional finite-difference time domain model (3D FDTD) has been developed to simulate acoustic wave interaction with a human head. A perfectly matched layer (PML) is used to absorb outgoing waves at the truncated boundary of an unbounded medium. An external source is utilized to reduce the computational domain size through the scattered field/total-field formulation. This numerical model has been validated by analytical solutions for a spherical head model. The 3D FDTD code is then used as a computational tool to predict the HRTF for various scenarios. In particular, a simplified spherical head model is compared to a realistic head model up to about 7 kHz. The HRTF is also computed for a realistic head model in the presence of a wall. It is demonstrated that this 3D FDTD model can be a useful tool for spatial audio applications. PMID- 12765363 TI - Antiplane shear wave propagation in fiber-reinforced composites. AB - A self-consistent method for analyzing antiplane shear wave propagation in two dimensional inhomogeneous media is presented. For applications in the high frequency range, the self-consistent condition for the effective medium is solved being supplemented with the theory of quasidynamic effective density. Comparisons with other theoretical calculations and experimental data for fiber-reinforced composites demonstrate the merits of using the present method. PMID- 12765364 TI - Helmholtz and parabolic equation solutions to a benchmark problem in ocean acoustics. AB - The Helmholtz equation (HE) describes wave propagation in applications such as acoustics and electromagnetics. For realistic problems, solving the HE is often too expensive. Instead, approximations like the parabolic wave equation (PE) are used. For low-frequency shallow-water environments, one persistent problem is to assess the accuracy of the PE model. In this work, a recently developed HE solver that can handle a smoothly varying bathymetry, variable material properties, and layered materials, is used for an investigation of the errors in PE solutions. In the HE solver, a preconditioned Krylov subspace method is applied to the discretized equations. The preconditioner combines domain decomposition and fast transform techniques. A benchmark problem with upslope-downslope propagation over a penetrable lossy seamount is solved. The numerical experiments show that, for the same bathymetry, a soft and slow bottom gives very similar HE and PE solutions, whereas the PE model is far from accurate for a hard and fast bottom. A first attempt to estimate the error is made by computing the relative deviation from the energy balance for the PE solution. This measure gives an indication of the magnitude of the error, but cannot be used as a strict error bound. PMID- 12765365 TI - Power dissipation and time-averaged pressure in oscillating flow through a sudden area change. AB - Experiments on oscillating flow at the abrupt transition between a two dimensional channel and essentially infinite space are presented. It is shown that phenomena associated with the transition are functions of three independent dimensionless parameters including the dimensionless radius rounding the edge of the end of the channel. The effect of each of these three parameters on the time averaged pressure difference across the transition and the acoustic power dissipation is explored by holding two parameters fixed while varying the third. Evidence is presented that the losses due to oscillatory flow in this geometry are smaller than would be expected from commonly accepted values for steady flow in similar geometry. PMID- 12765366 TI - Noncontacting lateral transportation using gas squeeze film generated by flexural traveling waves--numerical analysis. AB - This paper presents the theory describing the dynamical behavior of a noncontacting lateral transportation of planer objects by means of a gas squeeze film created by traveling flexural waves of a driving surface. An oscillating motion in the normal direction between two surfaces can generate a gas film with an average pressure higher than the surrounding. This load-carrying phenomenon arises from the fact that a viscous flow cannot be instantaneously squeezed; therefore, fast vibrations give rise to a cushioning effect. Equilibrium is established through a balance between viscous flow forces and compressibility forces. When the oscillatory motion between two surfaces creates traveling waves, lateral viscous forces are generated in addition to the normal levitation forces. These forces are produced as a result of nonuniform pressure gradients in the lateral direction between the surfaces. The combination of normal and lateral forces could be used for transporting objects without any direct contact with the driving surface. The numerical algorithm in this work couples the squeeze film phenomenon, which is represented by means of finite difference equations, to model a variant of the Reynolds equation, together with the equations describing the dynamics of the floating object. Numerical simulations are presented and investigated to highlight noteworthy topics. PMID- 12765367 TI - An optical fiber infrasound sensor: a new lower limit on atmospheric pressure noise between 1 and 10 Hz. AB - A new distributed sensor for detecting pressure variations caused by distant sources has been developed. The instrument reduces noise due to air turbulence in the infrasound band by averaging pressure along a line by means of monitoring strain in a long tubular diaphragm with an optical fiber interferometer. Above 1 Hz, the optical fiber infrasound sensor (OFIS) is less noisy than sensors relying on mechanical filters. Records collected from an 89-m-long OFS indicate a new low noise limit in the band from 1 to 10 Hz. Because the OFIS integrates pressure variations at light-speed rather than the speed of sound, phase delays of the acoustical signals caused by the sensor are negligible. Very long fiber-optic sensors are feasible and hold the promise of better wind-noise reduction than can be achieved with acoustical-mechanical systems. PMID- 12765368 TI - Diffraction of sound from a dipole source near to a barrier or an impedance discontinuity. AB - Pierce's formulation for the diffraction of spherical waves by a hard wedge has been extended to the case of the sound field due to a dipole source. The same approach is also used to extend a semiempirical model for sound propagation above an impedance discontinuity due to a dipole source. The resulting formulas have been validated by comparing their numerical solutions with that computed by summing the sound fields due to two closely spaced monopole sources of equal magnitude but opposite in phase. These new formulations are then used to develop a simple model for calculating the dipole sound field diffracted by a barrier above an impedance ground. Applications of these models relate to transportation noise prediction, particularly railway noise abatement, for which dipole sources are commonly used. The numerical predictions have been found to compare reasonably well with indoor measurements using piezoceramic transducers as dipole sources. PMID- 12765369 TI - Observations of acoustic surface waves in outdoor sound propagation. AB - Acoustic surface waves have been detected propagating outdoors under natural conditions. Two critical experimental conditions were employed to ensure the conclusive detection of these waves. First, acoustic pulses rather than a continuous wave source allowed an examination of the waveform shape and avoided the masking of wave arrivals. Second, a snow cover provided favorable ground impedance conditions for surface waves to exist. The acoustic pulses were generated by blank pistol shots fired 1 m above the snow. The resultant waveforms were measured using a vertical array of six microphones located 60 m away from the source at heights between 0.1 and 4.75 m. A strong, low frequency "tail" following the initial arrival was recorded near the snow surface. This tail, and its exponential decay with height (z) above the surface (approximately e(-alpha z)), are diagnostic features of surface waves. The measured attenuation coefficient alpha was 0.28 m(-1). The identification of the surface wave is confirmed by comparing the measured waveforms with waveforms predicted by the theoretical evaluation of the explicit surface wave pole term using residue theory. PMID- 12765371 TI - Coupled perturbed modes and internal solitary waves. AB - Coupled perturbed mode theory combines conventional coupled modes and perturbation theory. The theory is used to directly calculate mode coupling in a range-dependent shallow water problem involving propagation through continental shelf internal solitary waves. The solitary waves considered are thermocline depressions, separating well-mixed upper and lower layers. The method is fast and accurate. Results highlight mode coupling associated with internal solitary waves, and mode capture or loss to and from the discrete mode spectrum. PMID- 12765370 TI - Backscatter from a limestone seafloor at 2-3.5 kHz: measurements and modeling. AB - Physics-based interface scattering models for the seafloor [H.-H. Essen, J. Acoust. Soc. Am. 95, 1299-1310 (1994); Gragg et al., ibid. 110, 2878-2901 (2001)] exhibit features in their predicted grazing angle dependence. These features have a strong dependence on the assumed composition and roughness of the bottom. Verifying such predictions requires data that cover a wide range of grazing angles and involve minimal sub-bottom penetration. Such measurements were performed in the frequency band 2-3.5 kHz over an exposed limestone bottom off the Carolina coast during the second Littoral Warfare Advanced Development Focused Technology Experiment of 1996 (LWAD FTE 96-2). Direct-path bottom scattering strengths were obtained in shallow water (198-310 m deep) for grazing angles from 8 degrees to 75 degrees using data fusion from multiple experimental geometries coupled with careful signal processing. The processing included corrections for the surface-reflected path, other multipaths, and characteristics of the reverberation decay observed over the pulse duration at higher grazing angles. The resulting frequency and grazing-angle dependences exhibit trends consistent with theoretical predictions, and geoacoustic parameters obtained by inversion are consistent with values expected for limestone. PMID- 12765372 TI - Ray travel times at long ranges in acoustic waveguides. AB - The Hamiltonian formalism in terms of the action-angle variables is applied to study ray travel times in a waveguide with a smooth sound speed profile perturbed by a weak range-dependent inhomogeneity. A simple approximate formula relating the differences in ray travel times to range variations of action variables is derived. This relation is applied to study range variations of the timefront (representing ray arrivals in the time-depth plane). Widening and bias of timefront segments in the presence of perturbations are considered. Qualitative and quantitative explanations are given to surprising stability of early portions of timefronts observed in both numerical simulations and field experiments. This phenomenon is interpreted from the viewpoint of Fermat's principle. By ray tracing in a realistic deep water environment with an internal-wave-induced perturbation it has been demonstrated that our approach can be used at ranges up to, at least, 3000 km. PMID- 12765373 TI - Ray dynamics in long-range deep ocean sound propagation. AB - Recent results relating to ray dynamics in ocean acoustics are reviewed. Attention is focused on long-range propagation in deep ocean environments. For this class of problems, the ray equations may be simplified by making use of a one-way formulation in which the range variable appears as the independent (timelike) variable. Topics discussed include integrable and nonintegrable ray systems, action-angle variables, nonlinear resonances and the KAM theorem, ray chaos, Lyapunov exponents, predictability, nondegeneracy violation, ray intensity statistics, semiclassical breakdown, wave chaos, and the connection between ray chaos and mode coupling. The Hamiltonian structure of the ray equations plays an important role in all of these topics. PMID- 12765374 TI - The influence of large-scale seafloor slope and average bottom sound speed on low grazing-angle monostatic acoustic scattering. AB - Variations in large-scale seafloor slope and average seabed sound speed account for a significant portion of the variations in scattering intensity observed in low-grazing-angle monostatic reverberation. Numerical modeling using a finite difference solution to the elastic wave equation is used to quantify the effect of these large-scale parameters in interpretations of reverberation data. For hard rough seafloor (e.g., basalt), the results of the modeling suggest that the monostatic backscattering strength increases with increasing large-scale seafloor slope up to a slope of about 15 degrees dipping toward the incident direction. Once the grazing angle of the incident wavefield exceeds the critical grazing angle for the flat reference seafloor the backscattering intensity increases only slowly with increasing grazing angle. Similarly, average subseafloor sound speed has a significant effect. Seafloor with low sound speeds characteristic of soft bottoms (e.g., sediment) generate significantly weaker backscatter signals than seafloor with sound speeds characteristic of hard bottoms (e.g., basalt). The difference is that the shear waves can always be passed into soft bottoms because even for a flat seafloor there is no shear wave critical grazing angle. PMID- 12765375 TI - Microseism and infrasound generation by cyclones. AB - A two-dimensional cylindrical shear-flow wave theory for the generation of microseisms and infrasound by hurricanes and cyclones is developed as a linearized theory paralleling the seminal work by Longuet-Higgins which was limited to one-dimensional plane waves. Both theories are based on Bernoulli's principle. A little appreciated consequence of the Bernoulli principle is that surface gravity waves induce a time dependent pressure on the sea floor through a vertical column of water. A significant difference exists between microseisms detected at the bottom of each column and seismic signals radiated into the crust through coherence over a region of the sea floor. The dominant measured frequency of radiated microseisms is matched by this new theory for seismic data gathered at the Fordham Seismic Station both for a hurricane and a mid-latitude cyclone in 1998. Implications for Bernoulli's principle and this cylindrical stress flow theory on observations in the literature are also discussed. PMID- 12765376 TI - An iterative implementation of rotated coordinates for inverse problems. AB - A generalized inversion method is presented that uses a rotated coordinates technique [Collins and Fishman, J. Acoust. Soc. Am. 98, 1637-1644 (1995)] in simulated annealing to invert for both the location of an acoustic source and parameters that describe the ocean seabed. The rotated coordinates technique not only aids in the inversion process but also indicates the coupling of the source and environmental parameters and the relative sensitivities of the cost function to changes in the various parameters. The information obtained from the rotated coordinates provides insights into how the inversion problem can be effectively decoupled. An iterative process consisting of multiple simulated annealing runs that each use a different set of rotated coordinates is demonstrated. This multistep algorithm is called systematic decoupling using rotated coordinates and is especially helpful when inverting for a large number of unknown parameters. The cost function minimized in the inversion algorithm is model-data cross hydrophone spectra summed coherently over frequency and receiver pairs. The results of applying this inversion method to simulated data are presented in this paper. PMID- 12765377 TI - Broadband matched-field processing: coherent and incoherent approaches. AB - Matched-field based methods always involve the comparison of the output of a physical model and the actual data. The method of comparison and the nature of the data varies according to the problem at hand, but the result becomes always largely conditioned by the accurateness of the physical model and the amount of data available. The usage of broadband methods has become a widely used approach to increase the amount of data and to stabilize the estimation process. Due to the difficulties to accurately predict the phase of the acoustic field the problem whether the information should be coherently or incoherently combined across frequency has been an open debate in the last years. This paper provides a data consistent model for the observed signal, formed by a deterministic channel structure multiplied by a perturbation random factor plus noise. The cross frequency channel structure and the decorrelation of the perturbation random factor are shown to be the main causes of processor performance degradation. Different Bartlett processors, such as the incoherent processor [Baggeroer et al., J. Acoust. Soc. Am. 80, 571-587 (1988)], the coherent normalized processor [Z.-H. Michalopoulou, IEEE J. Ocean Eng. 21, 384-392 (1996)] and the matched phase processor [Orris et al., J. Acoust. Soc. Am. 107, 2563-2375 (2000)], are reviewed and compared to the proposed cross-frequency incoherent processor. It is analytically shown that the proposed processor has the same performance as the matched-phase processor at the maximum of the ambiguity surface, without the need for estimating the phase terms and thus having an extremely low computational cost. PMID- 12765378 TI - Material property estimation in thin plates using focused, synthetic-aperture acoustic beams. AB - The method developed here exploits the wide angular range of focused acoustic probes and the large synthetic aperture of scanned transducers to permit a rapid and reliable estimation of material properties in thin plates. It is found in several tests with various materials that estimates of elastic behavior using this method agree with contact measurements to within less than 5%. The method utilizes transmission (or reflection) coefficient reconstruction for an infinite thin plate, across a wide range of frequency and wave number, from which elastic property estimates are made. Data collected over a large synthetic acoustic aperture are processed with temporal and spatial Fourier transforms applied to change the acquired data from the coordinate and time domains to the wave number and frequency domains. Extrinsic real-beam effects on the data are accounted for with a complex transducer point analysis. Transmission measurements yield reconstructed data extending to the mode cutoffs, permitting easy and nearly unambiguous estimation of a subset of the elastic stiffnesses. For anisotropic plates, elastic stiffnesses are estimated with an inversion procedure that uses only limited data carefully selected from different portions of the measured scattering coefficient. Estimates are made by reconstructing in a stepwise fashion, based on sensitivity studies, where only one stiffness is estimated from the data at any one time, restricting the optimization to a robust one dimensional search. PMID- 12765379 TI - Elastic wave thermal fluctuations, ultrasonic waveforms by correlation of thermal phonons. AB - It is widely recognized that acoustic degrees of freedom coupled to a thermal bath have amplitudes which fluctuate with a mean square proportional to temperature; this is the basis for the Debye theory of the heat capacity of insulating solids. It is shown here that these elastic wave thermal phonons have correlation functions identical to the system's ultrasonic Green's function, and furthermore that thermal noise in ultrasonic detectors should have correlation functions equivalent to conventional waveforms obtained by active transmission and reception. This suggests the possibility of doing ultrasonics without a source. Theory for the identity is presented, and several room temperature laboratory confirmations are conducted in the frequency range 0.1-1.0 MHz. The thermal nature of the origin of these correlations is established by comparing their strength with theoretical expectations. Applications are discussed. PMID- 12765380 TI - Guided waves propagating in sandwich structures made of anisotropic, viscoelastic, composite materials. AB - The propagation of Lamb-like waves in sandwich plates made of anisotropic and viscoelastic material layers is studied. A semi-analytical model is described and used for predicting the dispersion curves (phase velocity, energy velocity, and complex wave-number) and the through-thickness distribution fields (displacement, stress, and energy flow). Guided modes propagating along a test-sandwich plate are shown to be quite different than classical Lamb modes, because this structure does not have the mirror symmetry, contrary to most of composite material plates. Moreover, the viscoelastic material properties imply complex roots of the dispersion equation to be found that lead to connections between some of the dispersion curves, meaning that some of the modes get coupled together. Gradual variation from zero to nominal values of the imaginary parts of the viscoelastic moduli shows that the mode coupling depends on the level of material viscoelasticity, except for one particular case where this phenomenon exists whether the medium is viscoelastic or not. The model is used to quantify the sensitivity of both the dispersion curves and the through-thickness mode shapes to the level of material viscoelasticity, and to physically explain the mode coupling phenomenon. Finite element software is also used to confirm results obtained for the purely elastic structure. Finally, experiments are made using ultrasonic, air-coupled transducers for generating and detecting guided modes in the test-sandwich structure. The mode-coupling phenomenon is then confirmed, and the potential of the air-coupled system for developing single-sided, contactless, NDT applications of such structures is discussed. PMID- 12765381 TI - Approximation of the Struve function H1 occurring in impedance calculations. AB - The problem of the rigid-piston radiator mounted in an infinite baffle has been studied widely for tutorial as well as for practical reasons. The resulting theory is commonly applied to model a loudspeaker in the audio-frequency range. A special function, the Struve function H1 (z), occurs in the expressions for the rigid-piston radiator. This Struve function is not readily available in programs such as Matlab or Mathcad, nor in computer languages such as FORTRAN and C. Therefore a simple and effective approximation of H1 (z) which is valid for all z is developed. Some examples of the application of the Struve function in acoustics are presented. PMID- 12765382 TI - Energy analysis of a piezoelectric body under nonuniform deformation. AB - One of the most powerful and clear methods for solving electromechanical transducer problems is the energy method based on the use of the Euler-Lagrange equations. The general expression is developed in a form convenient for applying the energy method to the calculation of the internal energy of a piezoelectric body under nonuniform deformation. The electrical and mechanical variables in this expression are separable under certain conditions and the underlying physics is illustrated with particular examples of bars made of piezoelectric ceramic for the case of transverse and axial polarization. In the case that the electrical and mechanical variables are not separable, the contribution of the mutual energy term to the total internal energy is expressed analytically. PMID- 12765383 TI - Flexural waves on narrow plates. AB - Flexural wave speeds on beams or plates depend upon the bending stiffnesses which differ by the well-known factor (1 - nu2). A quantitative analysis of a plate of finite lateral width displays the plate-to-beam transition, and permits asymptotic analysis that shows the leading order dependence on the width. Orthotropic plates are analyzed using both the Kirchhoff and Kirchhoff-Rayleigh theories, and isotropic plates are considered for Mindlin's theory with and without rotational inertia. A frequency-dependent Young's modulus for beams or strips of finite width is suggested, although the form of the correction to the modulus is not unique and depends on the theory used. The sign of the correction for the Kirchhoff theory is opposite to that for the Mindlin theory. These results indicate that the different plate and beam theories can produce quite distinct behavior. This divergence in predictions is further illustrated by comparison of the speeds for antisymmetric flexural, or torsional, modes on narrow plates. The four classical theories predict limiting wave speeds as the plate width vanishes, but the values are different in each case. The deviations can be understood in terms of torsional waves and how each theory succeeds, or fails, in approximating the effect of torsion. Dispersion equations are also derived, some for the first time, for the flexural edge wave in each of the four "engineering" theories. PMID- 12765384 TI - Negative group velocity Lamb waves on plates and applications to the scattering of sound by shells. AB - Symmetric Lamb waves on plates exhibit anomalies for certain regions of frequency. The phase velocity appears to be double-valued [M. F. Werby and H. Uberall, J. Acoust. Soc. Am. 111, 2686-2691 (2002)] with one of the branches having a negative group velocity relative to the corresponding phase velocity. The classification of the symmetric plate modes for frequencies appearing to have a double-valued phase velocity is reviewed here. The complication of a double valued velocity is avoided by examining mode orthogonality and the complex wave number spectra. Various authors have noted an enhancement in the backscattering of sound by elastic shells in water that occurs for frequencies where symmetric leaky Lamb waves (generalized to case of a shell) have contra-directed group and phase velocities. The ray diagram for negative group velocity contributions to the scattering by shells [G. Kaduchak, D. H. Hughes, and P. L. Marston, J. Acoust. Soc. Am. 96, 3704-3714 (1994)] is unusual since for this type of mode the energy on the shell flows in the opposite direction of the wave vector. Circumnavigation of the shell is not required for the leaky ray to be backward directed. PMID- 12765385 TI - Radiation efficiency of convected fluid-loaded plate. AB - The radiation of sound from geometrically simple vibrating structures into stationary fluids is well understood. However, to date, very few investigations have considered the effects of fluid convection on structural acoustic radiation. The purpose of this investigation is to quantify the effects that fluid flow has on the sound radiated from rectangular vibrating plates. The discussion includes a description of the fundamental physics associated with a simply supported, vibrating, rectangular plate imbedded in an infinite baffle and radiating into a semi-infinite convected fluid field. This is followed by a discussion of the computational approach used to calculate the plate radiation efficiency. Finally, numerical results are presented which demonstrate the effect that convection has on the radiation efficiency. The primary effect is a significant increase in radiation efficiency in the mid-wave number region, which is attributable to an effective decrease in the critical frequency. PMID- 12765386 TI - Solving the hypersingular boundary integral equation in three-dimensional acoustics using a regularization relationship. AB - Regularization of the hypersingular integral in the normal derivative of the conventional Helmholtz integral equation through a double surface integral method or regularization relationship has been studied. By introducing the new concept of discretized operator matrix, evaluation of the double surface integrals is reduced to calculate the product of two discretized operator matrices. Such a treatment greatly improves the computational efficiency. As the number of frequencies to be computed increases, the computational cost of solving the composite Helmholtz integral equation is comparable to that of solving the conventional Helmholtz integral equation. In this paper, the detailed formulation of the proposed regularization method is presented. The computational efficiency and accuracy of the regularization method are demonstrated for a general class of acoustic radiation and scattering problems. The radiation of a pulsating sphere, an oscillating sphere, and a rigid sphere insonified by a plane acoustic wave are solved using the new method with curvilinear quadrilateral isoparametric elements. It is found that the numerical results rapidly converge to the corresponding analytical solutions as finer meshes are applied. PMID- 12765387 TI - Evaluation of decay times in coupled spaces: Bayesian decay model selection. AB - This paper applies Bayesian probability theory to determination of the decay times in coupled spaces. A previous paper [N. Xiang and P. M. Goggans, J. Acoust. Soc. Am. 110, 1415-1424 (2001)] discussed determination of the decay times in coupled spaces from Schroeder's decay functions using Bayesian parameter estimation. To this end, the previous paper described the extension of an existing decay model [N. Xiang, I. Acoust. Soc. Am. 98, 2112-2121 (1995)] to incorporate one or more decay modes for use with Bayesian inference. Bayesian decay time estimation will obtain reasonable results only when it employs an appropriate decay model with the correct number of decay modes. However, in architectural acoustics practice, the number of decay modes may not be known when evaluating Schroeder's decay functions. The present paper continues the endeavor of the previous paper to apply Bayesian probability inference for comparison and selection of an appropriate decay model based upon measured data. Following a summary of Bayesian model comparison and selection, it discusses selection of a decay model in terms of experimentally measured Schroeder's decay functions. The present paper, along with the Bayesian decay time estimation described previously, suggests that Bayesian probability inference presents a suitable approach to the evaluation of decay times in coupled spaces. PMID- 12765388 TI - An acoustic-logging transmission-network model (continued): addition and multiplication ALTNs. AB - On the basis of the acoustic-logging transmission-network (ALTN) model [Fa et al., J. Acoust. Soc. Am. 111(5), 2158-2165 (2002)] this paper puts forward concepts of addition and multiplication ALTNs, proves the reciprocity of two kinds of addition ALTNs, sets up the physical model of the multiplication acoustic array logging tool, calculates their acoustic-beam steering efficiencies, and performs experimental verification. PMID- 12765389 TI - Performance bounds for passive sensor arrays operating in a turbulent medium: plane-wave analysis. AB - The performance bounds of a passive acoustic array operating in a turbulent medium with fluctuations described by a von Karman spectrum are investigated. This treatment considers a single, monochromatic, plane-wave source at near normal incidence. A line-of-sight propagation path is assumed. The primary interests are in calculating the Cramer-Rao lower bounds of the azimuthal and elevational angles of arrival and in observing how these bounds change with the introduction of additional unknowns, such as the propagation distance, turbulence parameters, and signal-to-noise ratio. In both two and three dimensions, it is found that for large values of the index-of-refraction variance, the Cramer-Rao lower bounds of the angles of arrival increase significantly at large values of the normalized propagation distance. For small values of the index-of-refraction variance and normalized propagation distance, the signal-to-noise ratio is found to be the limiting factor. In the two-dimensional treatment, it is found that the estimate of the angle of arrival will decouple from the estimates of the other parameters with the appropriate choice of array geometry. In three dimensions, again with an appropriate choice of array geometry, the estimates of the azimuth and elevation will decouple from the estimates of the other parameters, but due to the constraints of the model, will remain coupled to one another. PMID- 12765390 TI - Source motion mitigation for adaptive matched field processing. AB - Application of adaptive matched field processing to the problem of detecting quiet targets in shallow water is complicated by source motion, both the motion of the target and the motion of discrete interferers. Target motion causes spreading of the target peak, thereby reducing output signal power. Interferer motion increases the dimensionality of the interference subspace, reducing adaptive interference suppression. This paper presents three techniques that mitigate source motion problems in adaptive matched field processing. The first involves rank reduction, which enables adaptive weight computation over short observation intervals where motion effects are less pronounced. The other two techniques specifically compensate for source motion. Explicit target motion compensation reduces target motion mismatch by focusing snapshots according to a target velocity hypothesis. And time-varying interference filtering places time varying nulls on moving interferers not otherwise suppressed by adaptive weights. The three techniques are applied to volumetric array data from the Santa Barbara Channel Experiment and are shown to improve output signal-to-background-plus noise ratio by more than 3 dB over the standard minimum-variance, distortionless response adaptive beam-former. Application of the techniques in some cases proves to be the difference between detecting and not detecting the target. PMID- 12765391 TI - Optimal detection of crack echo families in elastic solids. AB - Optimal detection of a striplike crack residing in an isotropic elastic solid with coarse microstructure by means of ultrasonic nondestructive evaluation (NDE) is considered. A physics-based approach to derive an optimal detector, which achieves the theoretical limitations constrained by the underlying physics, is presented. State-of-the-art physical models of crack echoes and of stochastic backscattering from the material structure in elastic solids are introduced and unified with the theory of optimal detection to yield a practically useful nonlinear filter bank implementation of the optimal detector. Monte Carlo simulations of the detection performance for the special case of a striplike crack with uncertain angular orientation are presented in the form of receiver operating characteristics (ROCs). These new results represent the physical limitations for detecting a crack under the stated conditions and serve as performance bounds to which other detectors should be compared. A physics-based generalized likelihood ratio (GLR) detector, which relies on the same nonlinear filter bank as the optimal detector, is also presented for the special case of a striplike crack. A comparison between the optimal and the GLR detectors shows that the GLR detector only slightly reduces the performance. PMID- 12765392 TI - Transmission mode time-reversal super-resolution imaging. AB - The theory of time-reversal super-resolution imaging of point targets embedded in a reciprocal background medium [A. J. Devaney, "Super-resolution imaging using time-reversal and MUSIC," J. Acoust. Soc. Am. (to be published)] is generalized to the case where the transmitter and receiver sensor arrays need not be coincident and for cases where the background medium can be nonreciprocal. The new theory developed herein is based on the singular value decomposition of the generalized multistatic data matrix of the sensor system rather than the standard eigenvector/eigenvalue decomposition of the time-reversal matrix as was employed in the above-mentioned work and other treatments of time-reversal imaging [Prada, Thomas, and Fink, "The iterative time reversal process: Analysis of the convergence," J. Acoust. Soc. Am. 97, 62 (1995); Prada et al., "Decomposition of the time reversal operator: Detection and selective focusing on two scatterers," J. Acoust. Soc. Am. 99, 2067 (1996)]. A generalized multiple signal classification (MUSIC) algorithm is derived that allows super-resolution imaging of both well-resolved and non-well-resolved point targets from arbitrary sensor array geometries. MUSIC exploits the orthogonal nature of the scatterer and noise subspaces defined by the singular vectors of the multistatic data matrix to form scatterer images. The time-reversal/MUSIC algorithm is tested and validated in two computer simulations of offset vertical seismic profiling where the sensor sources are aligned along the earth's surface and the receiver array is aligned along a subsurface borehole. All results demonstrate the high contrast, high resolution imaging capabilities of this new algorithm combination when compared with "classical" backpropagation or field focusing. Above and beyond the application of seismo-acoustic imaging, the time-reversal super-resolution theory has applications in ocean acoustics for target location, and ultrasonic nondestructive evaluation of parts. PMID- 12765393 TI - Reciprocal maximum-length sequence pairs for acoustical dual source measurements. AB - In this paper we propose and demonstrate a method to obtain simultaneous dual source-receiver impulse responses in acoustical systems using binary maximum length sequences (MLS). A binary MLS and its reversed-order sequence form a reciprocal MLS pair. Their correlation property includes a two-valued "pulse like" autocorrelation function and a relatively smaller-valued cross-correlation function. This unique property, along with other number-theory properties, makes the reciprocal MLS pair suitable for simultaneous dual source cross-correlation measurements. In the measurement of a dual source system, each of the reciprocal MLS pairs simultaneously excite one of two separate sources, one or several receiver signals cross-correlate in turn with each of the MLS pairs, resulting in impulse responses associated with two separate sources. The proposed method is particularly valuable for system identification tasks with multiple sound/vibration sources and receivers that have to be accomplished in a limited time period. A fast algorithm called a fast MLS transform is exploited for the cross-correlation. In this paper we propose a fast MLS transform pair for the reciprocal MLS pairs. Its efficiency lies in the requirement of one single permutation matrix for a pair of two fast MLS transforms. Its feasibility and usefulness in the acoustical measurements are demonstrated using experimental results. PMID- 12765394 TI - Stimulus-frequency-emission group delay: a test of coherent reflection filtering and a window on cochlear tuning. AB - This paper tests and applies a key prediction of the theory of coherent reflection filtering for the generation of reflection-source otoacoustic emissions. The theory predicts that reflection-source-emission group delay is determined by the group delay of the basilar-membrane (BM) transfer function at its peak. This prediction is tested over a seven-octave frequency range in cats and guinea pigs using measurements of stimulus-frequency-emission (SFOAE) group delay. A comparison with group delays calculated from published measurements of BM mechanical transfer functions supports the theory at the basal end of the cochlea. A comparison across the whole frequency range based on variations in the sharpness of neural tuning with characteristic frequency (CF) suggests that the predicted relation holds in the basal-most 60% of the cochlea. At the apical end of the cochlea, however, the measurements disagree with neural and mechanical group delays. This disagreement suggests that there are important differences in cochlear mechanics and/or mechanisms of emission generation between the base and apex of the cochlea. Measurements in humans over a four-octave range indicate that human SFOAE group delays are roughly a factor of 3 longer than their counterparts in cat and guinea pig but manifest similar trends across CF. The measurements thus reveal global deviations from scaling whose form appears quantitatively similar in all three species. Interpreted using the theory of coherent reflection filtering, the group delay measurements indicate that the wavelength at the peak of the traveling wave decreases with increasing CF at a rate of roughly 25% per octave in the base of the cochlea. The measurements and analysis reported here illustrate the rich potential inherent in OAE measurements for obtaining valuable information about basic cochlear properties such as tuning. PMID- 12765395 TI - Measurements of human middle ear forward and reverse acoustics: implications for otoacoustic emissions. AB - Middle and inner ears from human cadaver temporal bones were stimulated in the forward direction by an ear-canal sound source, and in the reverse direction by an inner-ear sound source. For each stimulus type, three variables were measured: (a) Pec--ear-canal pressure with a probe-tube microphone within 3 mm of the eardrum, (b) Vst--stapes velocity with a laser interferometer, and (c) Pv- vestibule pressure with a hydrophone. From these variables, the forward middle ear pressure gain (M1), the cochlear input impedance (Zc), the reverse middle-ear pressure gain (M2), and the reverse middle-ear impedance (M3) are directly obtained for the first time from the same preparation. These measurements can be used to fully characterize the middle ear as a two-port system. Presently, the effect of the middle ear on otoacoustic emissions (OAEs) is quantified by calculating the roundtrip middle-ear pressure gain Gme(RT) as the product of M1 and M2. In the 2-6.8 kHz region, absolute value(Gme(RT)) decreases with a slope of -22 dB/oct, while OAEs (both click evoked and distortion products) tend to be independent of frequency; this suggests a steep slope in vestibule pressure from 2 kHz to at least 4 kHz for click evoked OAEs and to at least 6.8 kHz for distortion product OAEs. Contrary to common assumptions, measurements indicate that the emission generator mechanism is frequency dependent. Measurements are also used to estimate the reflectance of basally traveling waves at the stapes, and apically generated nonlinear reflections within the vestibule. PMID- 12765396 TI - Microsecond temporal resolution in monaural hearing without spectral cues? AB - The auditory system encodes the timing of peaks in basilar-membrane motion with exquisite precision, and perceptual models of binaural processing indicate that the limit of temporal resolution in humans is as little as 10-20 microseconds. In these binaural studies, pairs of continuous sounds with microsecond differences are presented simultaneously, one sound to each ear. In this paper, a monaural masking experiment is described in which pairs of continuous sounds with microsecond time differences were combined and presented to both ears. The stimuli were matched in terms of the excitation patterns they produced, and a perceptual model of monaural processing indicates that the limit of temporal resolution in this case is similar to that in the binaural system. PMID- 12765397 TI - Testing the concept of a modulation filter bank: the audibility of component modulation and detection of phase change in three-component modulators. AB - Two experiments were performed to test the concept that the auditory system contains a "modulation filter bank" (MFB). Experiment 1 examined the ability to "hear out" the modulation frequency of the central component of a three-component modulator applied to a 4-kHz sinusoidal carrier. On each trial, three modulated stimuli were presented. The modulator of the first stimulus contained three components. Within a run the frequencies of the outer two components were fixed and the frequency of the central ("target") component was drawn randomly from one of five values. The modulators of second and third stimuli contained one component. One had a frequency equal to that of the target and the other had a frequency randomly selected from one of the other possible values. Subjects indicated whether the target corresponded to the second or third stimulus. Scores were around 80% correct when the components in the three-component modulator were widely spaced and when the frequencies of the target and comparison differed sufficiently. Experiment 2 examined the ability to hear a change in the relative phase of the components in a three-component modulator with harmonically spaced components, using a 31FC task. The frequency of the central component, f(c), was either 50 or 100 Hz. Scores were 80%-90% correct when the component spacing was < or = 0.5 f(c), but decreased markedly for greater spacings. Performance was only slightly impaired by randomizing the overall modulation depth from one stimulus to the next. The results of both experiments are broadly consistent with what would be expected from a MFB with a Q value of 1 or slightly less. PMID- 12765398 TI - The relation of lung volume initiation to selected acoustic properties of speech. AB - This study examined the relationship of speech breathing to other elements of speech production. It was hypothesized that initiating speech from different lung volumes would have an effect on different elements of the acoustic output. It was postulated that effects may be brought about by mechanical interaction as well as a dispersion of effort to mechanically unlinked elements of speech production, such as articulatory behavior. To this end, selected acoustic variables were studied in eight young healthy women who initiated speech from low, typical, and high lung volume levels. The acoustic variables studied were selected because they have been shown to be sensitive indicators of speech production performance. It was found that with increasing lung volume initiation levels, average sound pressure level, average fundamental frequency, and declination rate of fundamental frequency increased. It was also observed that vowel space was significantly smaller during low lung volume initiation levels relative to typical lung volume initiation levels. Vowel space reduction is discussed relative to "gaining down." PMID- 12765400 TI - Effects of prosodic boundary on /aC/ sequences: articulatory results. AB - This study presents EMA (electromagnetic articulography) data on articulation of the vowel /a/ at different prosodic boundaries in French. Three speakers of metropolitan French produced utterances containing the vowel /a/, preceded by /t/ and followed by one of six consonants /b d g f s S/ (three stops and three fricatives), with different prosodic boundaries intervening between the /a/ and the six different consonants. The prosodic boundaries investigated are the Utterance, the Intonational phrase, the Accentual phrase, and the Word. Data for the Tongue Tip, Tongue Body, and Jaw are presented. The articulatory data presented here were recorded at the same time as the acoustic data presented in Tabain [J. Acoust. Soc. Am. 113, 516-531 (2003)]. Analyses show that there is a strong effect on peak displacement of the vowel according to the prosodic hierarchy, with the stronger prosodic boundaries inducing a much lower Tongue Body and Jaw position than the weaker prosodic boundaries. Durations of both the opening movement into and the closing movement out of the vowel are also affected. Peak velocity of the articulatory movements is also examined, and, contrary to results for phrase-final lengthening, it is found that peak velocity of the opening movement into the vowel tends to increase with the higher prosodic boundaries, together with the increased magnitude of the movement between the consonant and the vowel. Results for the closing movement out of the vowel and into the consonant are not so clear. Since one speaker shows evidence of utterance-level articulatory declension, it is suggested that the competing constraints of articulatory declension and prosodic effects might explain some previous results on phrase-final lengthening. PMID- 12765399 TI - Tongue-surface movement patterns during speech and swallowing. AB - The tongue has been frequently characterized as being composed of several functionally independent articulators. The question of functional regionality within the tongue was examined by quantifying the strength of coupling among four different tongue locations across a large number of consonantal contexts and participants. Tongue behavior during swallowing was also described. Vertical displacements of pellets affixed to the tongue were extracted from the x-ray microbeam database. Forty-six participants recited 20 vowel-consonant-vowel (VCV) combinations and swallowed 10 ccs of water. Tongue-surface movement patterns were quantitatively described by computing the covariance between the vertical time histories of all possible pellet pairs. Phonemic differentiation in vertical tongue motions was observed as coupling varied predictably across pellet pairs with place of articulation. Moreover, tongue displacements for speech and swallowing clustered into distinct groups based on their coupling profiles. Functional independence of anterior tongue regions was evidenced by a wide range of movement coupling relations between anterior tongue pellets. The strengths and weaknesses of the covariance-based analysis for characterizing tongue movement are considered. PMID- 12765401 TI - Using links between speech perception and speech production to evaluate different acoustic metrics: a preliminary report. AB - This paper examines whether correlations between speech perception and speech production exist, and, if so, whether they might provide a way of evaluating different acoustic metrics. The cues listeners use for many phonemic distinctions are not known, often because many different acoustic cues are highly correlated with one another, making it difficult to distinguish among them. Perception production correlations may provide a new means of doing so. In the present paper, correlations were examined between acoustic measures taken on listeners' perceptual prototypes for a given speech category and on their average production of members of that category. Significant correlations were found for VOT among stop consonants, and for spectral peaks (but not centroids or skewness) for voiceless fricatives. These results suggest that correlations between speech perception and production may provide a methodology for evaluating different proposed acoustic metrics. PMID- 12765402 TI - The resolution of complex spectral patterns by cochlear implant and normal hearing listeners. AB - The differences in spectral shape resolution abilities among cochlear implant (CI) listeners, and between CI and normal-hearing (NH) listeners, when listening with the same number of channels (12), was investigated. In addition, the effect of the number of channels on spectral shape resolution was examined. The stimuli were rippled noise signals with various ripple frequency-spacings. An adaptive 41FC procedure was used to determine the threshold for resolvable ripple spacing, which was the spacing at which an interchange in peak and valley positions could be discriminated. The results showed poorer spectral shape resolution in CI compared to NH listeners (average thresholds of approximately 3000 and 400 Hz, respectively), and wide variability among CI listeners (range of approximately 800 to 8000 Hz). There was a significant relationship between spectral shape resolution and vowel recognition. The spectral shape resolution thresholds of NH listeners increased as the number of channels increased from 1 to 16, while the CI listeners showed a performance plateau at 4-6 channels, which is consistent with previous results using speech recognition measures. These results indicate that this test may provide a measure of CI performance which is time efficient and non-linguistic, and therefore, if verified, may provide a useful contribution to the prediction of speech perception in adults and children who use CIs. PMID- 12765403 TI - Numerical and experimental modal analysis of the reed and pipe of a clarinet. AB - A modal computation of a complete clarinet is presented by the association of finite-element models of the reed and of part of the pipe with a lumped-element model of the rest of the pipe. In the first part, we compare modal computations of the reed and the air inside the mouthpiece and barrel with measurements performed by holographic interferometry. In the second part, the complete clarinet is modeled by adjoining a series of lumped elements for the remaining part of the pipe. The parameters of the lumped-resonator model are determined from acoustic impedance measurements. Computed eigenmodes of the whole system show that modal patterns of the reed differ significantly whether it is alone or coupled to air. Some modes exhibit mostly reed motion and a small contribution of the acoustic pressure inside the pipe. Resonance frequencies measured on a clarinet with the mouthpiece replaced by the cylinder of equal volume differ significantly from the computed eigenfrequencies of the clarinet taking the actual shape of the mouthpiece into account and from those including the (linear) dynamics of the reed. This suggests revisiting the customary quality index based on the alignment of the peaks of the input acoustical impedance curve. PMID- 12765404 TI - Ultrasonic absorption in aqueous solutions of amino acids at neutral pH. AB - Ultrasonic absorption coefficients in aqueous solutions of glycine, L-alanine, imidazole, L-phenylalanine, L-histidine and L-tryptophan at neutral pH were measured in the range from 0.8 to 220 MHz at 25 degrees C. A characteristic ultrasonic relaxation phenomenon was observed only in the solution of L-histidine with a relaxation frequency at around 2 MHz at neutral pH. It was proposed from the concentration independent relaxation frequency and the linear concentration dependence of the maximum absorption per wavelength that the relaxation mechanism was associated with a perturbation of the rotational isomeric equilibrium of the L-histidine molecule. The existence of two rotational isomeric forms of L histidine in water was examined by semiempirical quantum chemical methods, in order to determine the free energy difference between the two states. The forward and backward rate constants were determined from the relaxation frequency and the energy change. Also, the standard volume change of the reaction was estimated from the concentration dependence of the maximum absorption per wavelength. It was speculated that L-histidine fulfills a specific function among amino acids because of the rotational motion in the molecule, in addition to its well established acid-base properties. PMID- 12765405 TI - Multiple scattering in a trabecular bone: influence of the marrow viscosity on the effective properties. AB - The Foldy and the Waterman and Truell approximations are used to determine the effective properties of the coherent wave that emerges after multiple scattering of a plane longitudinal fast wave by the largest pores in a trabecular bone. The unit scattering cell considered is either a single pore or two close cylindrical pores (cluster), at a fixed overall bone porosity. In the cluster case, the effective attenuation is about twice that obtained with one single pore per scatterer. It is shown that taking into account the marrow viscosity leads only to minor differences on the effective dispersion and attenuation. PMID- 12765406 TI - Modeling the frequency dependence (5-120 MHz) of ultrasound backscattering by red cell aggregates in shear flow at a normal hematocrit. AB - The frequency dependence of the ultrasound signal backscattered by blood in shear flow was studied using a simulation model. The ultrasound backscattered signal was computed with a linear model that considers the characteristics of the ultrasound system and tissue acoustic properties. The tissue scattering properties were related to the position and shape of the red blood cells (RBCs). A 2D microrheological model simulated the RBC dynamics in a Couette shear flow system. This iterative model, described earlier [Biophys. J. 82, 1696-1710 (2002)], integrates the hydrodynamic effect of the flow, as well as adhesive and repulsive forces between RBCs. RBC aggregation was simulated at 40% hematocrit and shear rates of 0.05-2 s(-1). The RBC aggregate sizes ranged, on average, from 3.3 RBCs at 2 s(-1) to 33.5 cells at 0.05 s(-1). The ultrasound backscattered power was studied at frequencies between 5-120 MHz and insonification angles between 0-180 degrees. At frequencies below approximately 30 MHz, the ultrasound backscattered power increased as the shear rate was decreased and the size of the aggregates was raised. A totally different scattering behavior was noted above 30 MHz. Typical spectral slopes of the backscattered power (log-log scale) between 5 25 MHz equaled 3.8, whereas slopes down to 0.6 were measured at 0.05 s(-1), between 40-60 MHz. The ultrasound backscattered power was shown to be angle dependent at low frequencies (5-25 MHz). The anisotropy persisted at high frequencies (>25 MHz) for small aggregates (at 2 s(-1)). In conclusion, this study sheds some light on the blood backscattering behavior with an emphasis on the non-Rayleigh regime. Additional experimental studies may be necessary to validate the simulation results, and to fully understand the relation between the ultrasound backscattered power, level of RBC aggregation, shear rate, frequency, and insonification angle. PMID- 12765407 TI - Reliability of sound attenuation in Florida scrub habitat and behavioral implications. AB - Attenuation over distance in natural habitat is often difficult to predict when measured without respect to sound frequency. The physical-acoustic structure of Florida scrub habitat is described and both attenuation and reliability of attenuation are measured as a function of sound frequency, over several distances, speaker elevations, and microphone elevations. The spatial context of sound propagation in Florida scrub habitat is discussed and a model designed to describe contributions to overall attenuation from individual factors is presented. Sound frequencies below approximately 3.5 kHz attenuate more reliably than higher sound frequencies, suggesting that animals should pay greatest attention to relatively low sound frequencies when they assess attenuation or estimate sound-pressure level. PMID- 12765408 TI - Effect of pulse polarity and energy on ultrasound-induced lung hemorrhage in adult rats. AB - The objective of this study was to further assess the role of inertial cavitation in ultrasound-induced lung hemorrhage by examining the effect of pulse polarity at a common in situ (at the lung surface) peak rarefactional pressure [pr(in situ)] and at a common in situ pulse intensity integral (PII(in situ)). A total of 60 rats was divided into three experimental groups of 20 animals per group and randomly exposed to pulsed ultrasound. The groups were exposed as follows: Group 1 to 0 degree polarity pulses (compression followed by rarefraction) at a pr(in situ) of 3.48 MPa and a PII(in situ) of 4.78 Ws/m2, group 2 to 180 degree polarity pulses (rarefraction followed by compression) at a pr(in situ) of 3.72 MPa and a PII(in situ) of 2.55 Ws/m2, and group 3 to 180 degree polarity pulses at a pr(in situ) of 4.97 MPa and a PII(in situ) of 4.79 Ws/m2. For all experimental groups, the frequency was 2.46 MHz, the exposure duration was 240 s, the pulse repetition frequency was 2.5 kHz, and the pulse duration was 0.42 micros. Six sham animals were also randomly distributed among the experimental animals. The lesion surface area and depth were determined for each rat as well as lesion occurrence (percentage of rats with lesions) per group. It was found that lesion occurrence and size correlated better with PII(in situ) than with pr(in situ), suggesting that a mechanism other than inertial cavitation was responsible for the damage. PMID- 12765409 TI - Pulse register phonation in Diana monkey alarm calls. AB - The adult male Diana monkeys (Cercopithecus diana) produce predator-specific alarm calls in response to two of their predators, the crowned eagles and the leopards. The acoustic structure of these alarm calls is remarkable for a number of theoretical and empirical reasons. First, although pulsed phonation has been described in a variety of mammalian vocalizations, very little is known about the underlying production mechanism. Second, Diana monkey alarm calls are based almost exclusively on this vocal production mechanism to an extent that has never been documented in mammalian vocal behavior. Finally, the Diana monkeys' pulsed phonation strongly resembles the pulse register in human speech, where fundamental frequency is mainly controlled by subglottal pressure. Here, we report the results of a detailed acoustic analysis to investigate the production mechanism of Diana monkey alarm calls. Within calls, we found a positive correlation between the fundamental frequency and the pulse amplitude, suggesting that both humans and monkeys control fundamental frequency by subglottal pressure. While in humans pulsed phonation is usually considered pathological or artificial, male Diana monkeys rely exclusively on pulsed phonation, suggesting a functional adaptation. Moreover, we were unable to document any nonlinear phenomena, despite the fact that they occur frequently in the vocal repertoire of humans and nonhumans, further suggesting that the very robust Diana monkey pulse production mechanism has evolved for a particular functional purpose. We discuss the implications of these findings for the structural evolution of Diana monkey alarm calls and suggest that the restricted variability in fundamental frequency and robustness of the source signal gave rise to the formant patterns observed in Diana monkey alarm calls, used to convey predator information. PMID- 12765410 TI - Responses of squirrel monkeys to their experimentally modified mobbing calls. AB - Previous acoustic analyses suggested emotion-correlated changes in the acoustic structure of squirrel monkey (Saimiri sciureus) vocalizations. Specifically, calls given in aversive contexts were characterized by an upward shift in frequencies, often accompanied by an increase in amplitude. In order to test whether changes in frequencies or amplitude are indeed relevant for conspecific listeners, playback experiments were conducted in which either frequencies or amplitude of mobbing calls were modified. Latency and first orienting response were measured in playback experiments with six adult squirrel monkeys. After broadcasting yaps with increased frequencies or amplitude, squirrel monkeys showed a longer orienting response towards the speaker than after the corresponding control stimuli. Furthermore, after broadcasting yaps with decreased frequencies or amplitude, squirrel monkeys showed a shorter orienting response towards the speaker than after the corresponding manipulated calls with higher frequencies or amplitude. These results suggest that changes in frequencies or amplitude were perceived by squirrel monkeys, indicating that the relationship between call structure and the underlying affective state of the caller agreed with the listener's assessment of the calls. However, a simultaneous increase in frequencies and amplitude did not lead to an enhanced response, compared to each single parameter. Thus, from the receiver's perspective, both call parameters may mutually replace each other. PMID- 12765411 TI - Correlations between acoustic properties and bone density in bovine cancellous bone from 0.5 to 2 MHz. AB - Correlations between acoustic properties and bone density were investigated in the 12 defatted bovine cancellous bone specimens in vitro. Speed of sound (SOS) and broadband ultrasonic attenuation (BUA) were measured in three different frequency bandwidths from 0.5 to 2 MHz using three matched pairs of transducers with the center frequencies of 1, 2.25, and 3.5 MHz. The relative orientation between ultrasonic beam and bone specimen was the mediolateral (ML) direction of the bovine tibia. SOS shows significant linear positive correlation with apparent density for all three pairs of transducers. However, BUA shows relatively weak correlation with apparent density. SOS and BUA are only weakly correlated with each other. The linear combination of SOS and BUA in a multiple regression model leads to a significant improvement in predicting apparent density. The correlations among SOS, BUA, and bone density can be effectively and clearly represented in the three-dimensional space by the multiple regression model. These results suggest that the frequency range up to 1.5 MHz and the multiple regression model in the three-dimensional space can be useful in the osteoporosis diagnosis. PMID- 12765412 TI - Severe acute respiratory syndrome (SARS). PMID- 12765413 TI - Is Chlamydia pneumoniae an important pathogen in patients with community-acquired pneumonia? PMID- 12765414 TI - Tidal expiratory flow limitation, dyspnoea and exercise capacity in patients with bilateral bronchiectasis. AB - In this study the authors investigated whether expiratory flow limitation (FL) is present during tidal breathing in patients with bilateral bronchiectasis (BB) and whether it is related to the severity of chronic dyspnoea (Medical Research Council (MRC) dyspnoea scale), exercise capacity (maximal mechanical power output (WRmax)) and severity of the disease, as assessed by high-resolution computed tomography (HRCT) scoring. Lung function, MRC dyspnoea, HRCT score, WRmax and FL were assessed in 23 stable caucasian patients (six males) aged 56 +/- 17 yrs. FL was assessed at rest both in seated and supine positions. To detect FL, the negative expiratory pressure (NEP) technique was used. The degree of FL was rated using a five-point FL score. WRmax was measured using a cyclo-ergometer. According to the NEP technique, five patients were FL during resting breathing when supine but not seated, four were FL both seated and supine, and 14 were NFL both seated and supine. Furthermore, it was shown that: 1) in stable BB patients FL during resting breathing is common, especially in the supine position; 2) the degree of MRC dyspnoea is closely related to the five-point FL score; 3) WRmax (% pred) is more closely correlated with the MRC dyspnoea score than with the five point FL score; and 4) HRCT score is closely related to forced expiratory volume in one second % pred but not five-point FL score. In conclusion, flow limitation is common at rest in sitting and supine positions in patients with bilateral bronchiectasis. Flow limitation and reduced exercise capacity are both associated with more severe dyspnoea. Finally, high-resolution computed tomography scoring correlates best with forced expiratory volume in one second. PMID- 12765415 TI - Rigid bronchoscopy induces bacterial translocation: an experimental study in rats. AB - Bronchoscopy has the potential to propagate infections. Bacterial translocation was hypothesised to be the cause of infections observed following bronchoscopy and this study was designed to assess the risk of bacterial translocation following rigid bronchoscopy in rats. A total of 30 rats were evaluated. The study group (n=15) underwent rigid bronchoscopy. Arterial blood gas analysis was performed in all rats. Blood and tissue cultures from the ileum, caecum, mesenteric lymph nodes, liver, spleen, mediastinal lymph nodes and lung were obtained 24 h following bronchoscopy. Bacterial translocation to the mesenteric lymph nodes was found in seven of 15 rats (46.7%) that underwent bronchoscopy, compared with none of the controls. Of the seven positives, three rats (42.8%) also demonstrated other organ involvement, such as the liver and spleen. Escherichia coli, Salmonella typhymirium, S. enteritidis and Pseudomonas spp. were found as translocating bacteria. In the study group, pH and arterial oxygen tension were significantly lower and arterial carbon dioxide tension was higher, compared with controls. This study shows that rigid bronchoscopy may induce bacterial translocation in rats. Further investigations aimed at understanding the clinical consequences of this phenomenon are warranted. PMID- 12765416 TI - Fragile histidine triad protein expression in nonsmall cell lung cancer and correlation with Ki-67 and with p53. AB - Fragile histidine triad (FHIT) is a tumour suppressor gene, which is altered in a variety of epithelial tumours, including lung cancer. Biochemical and functional pathways of its tumourigenicity are not yet understood. Its role in tumour proliferation is particularly controversial. The purpose of this study was to correlate the expression of FHIT protein in nonsmall cell lung cancer (NSCLC) with tumour proliferation as estimated by Ki-67 antigen and with p53, a suppressor gene. FHIT, Ki-67 and p53 expression were evaluated by immunohistochemistry in 119 resected NSCLC. Altogether, 58 tumours were negative (expression <10%) for FHIT. The median expression in tumours was 15% positive cells, in comparison with 100% in normal matched lung tissue. The expression was as strong as in normal tissue in only 19 cases. FHIT expression was significantly lower in squamous cell carcinoma (SCC) (5%) than in adenocarcinoma (ADC) (64%). The median expression of Ki-67 was 20% and 69% of tumours were positives (expression >10%). Ki-67 expression was significantly higher in SCC (33.3%) than in ADC (10%). The loss of FHIT protein was not correlated with the expression of p53 (median: 7.5%, 58% of positive tumours for a cut-off of 10% of positive cells) or Ki-67. But percentage of labelled cells for p53 and Ki-67 were significantly correlated. The results suggest that for fragile histidine triad, the pathway of tumourigenesis is independent of p53 and of tumoural proliferation, as reported previously in vitro. PMID- 12765417 TI - Transcription factor activation and mitogenic synergism in airway smooth muscle cells. AB - Simultaneous treatment of human airway smooth muscle (HASM) cells with lysophosphatidic acid (LPA) and epidermal growth factor (EGF) leads to strikingly synergistic stimulation of mitogenesis. The purpose of this study was to explore potential sites for signal integration mediating synergism, focusing on extracellular signal-regulated kinase (ERK) and transcription factors involved in proliferation and inflammation as likely candidates. Activation of ERK was analysed by immunoblotting. Transcription factor activation was assessed using HASM cells transduced with luciferase reporter gene constructs. LPA and EGF both activated ERK but had no synergistic effect when combined. LPA and EGF both activated activator protein (AP)-1, cyclic adenosine monophosphate response element-binding protein, nuclear factor of activated T-cells and the serum response element; however, only AP-1 activation exhibited synergism. Activation of the inhibitory guanine nucleotide-binding protein and of ERK signalling pathways were required for most transcription factor responses to LPA. In contrast, nuclear factor (NF)-kappaB was activated by LPA but not EGF and NF kappaB activation was completely blocked only when Rho was inhibited. Rapid activation of Rho was observed in response to LPA but not to EGF. Importantly, inhibition of Rho selectively blocked synergism in both AP-1 activation and mitogenesis. In summary, extracellular signal-regulated kinase activation is required for many transcription factor responses to lysophosphatidic acid and epidermal growth factor, however it is not synergistic. Activation of activator protein-1 is synergistic, and Rho activation by lysophosphatidic acid is required for synergism in both activator protein-1 activation and mitogenesis. PMID- 12765418 TI - Increased eNO and pulmonary iNOS expression in eNOS null mice. AB - Nitric oxide (NO) is a major regulatory molecule of the cardiovascular system; however, measurement of vascular NO synthesis in vivo represents a major challenge. NO stemming from the lower respiratory tract has been used as a marker of vascular endothelial function. Experimental evidence for this concept is lacking. Therefore, the aim of the present study was to investigate this relationship. Lower respiratory tract exhaled NO concentration, together with systemic and pulmonary artery pressure, was measured in endothelial nitric oxide synthase (NOS) (eNOS) null mice (eNOS-/-). Similar studies were performed in inducible NOS (iNOS) null mice (iNOS-/-). Defective endothelial NO synthesis in eNOS-/- mice (evidenced by systemic and pulmonary hypertension) was associated with augmented exhaled NO levels (12.5 +/- 1.9 versus 9.8 +/- 1.2 parts per billion (ppb), eNOS-/- versus wild type), whereas normal endothelial NO synthesis in iNOS-/- mice was associated with decreased exhaled NO levels (4.3 +/- 1.5 ppb). Augmented exhaled NO levels in eNOS-/- mice were associated with upregulation of iNOS expression in the lung. These results indicate that inducible nitric oxide synthase is a major determinant of gaseous nitric oxide production in the lung, and lower respiratory tract exhaled nitric oxide does not always represent a marker of vascular endothelial nitric oxide synthesis. PMID- 12765419 TI - Increased vascular endothelial growth factor in acute eosinophilic pneumonia. AB - Acute eosinophilic pneumonia (AEP) is associated with the presence of diffuse pulmonary infiltrates on the chest radiograph and an increased number of eosinophils and an elevation of interleukin (IL)-5 levels in bronchoalveolar lavage (BAL) fluid. Vascular endothelial growth factor (VEGF) is a constitutively expressed protein encoded by messenger ribonucleic acid in human eosinophils and is released following stimulation with IL-5. However, the roles of IL-5 and VEGF in the pathogenesis or activity of this disease have not been clarified. The authors investigated the cells and the levels of these two factors in BAL fluid in five AEP patients and five normal controls before and after corticosteroid treatment. The absolute number of eosinophils-mL(-1), IL-5 and VEGF levels in patients before treatment were higher than in controls (53.8 versus 0.3 x 10(4) x mL(-1), 490.1 versus 5.2 pg x mL(-1) and 643.0 versus 133.9 pg x mL(-1), respectively). IL-5 and VEGF rapidly decreased to the control level in parallel with clinical improvement. The relationship between eosinophilia and IL-5 and VEGF levels was strongly significant. Elevated interleukin-5 in the lung may initiate the recruitment of eosinophils and enhance the release of mediators, such as vascular endothelial growth factor from eosinophils, which, in turn, increases the permeability of blood vessels. PMID- 12765420 TI - Chlamydia species as a cause of community-acquired pneumonia in Canada. AB - Chlamydia pneumoniae has been implicated as a cause of community-acquired pneumonia (CAP) in several studies. However, there has been no comprehensive study of the role of Chlamydia species (C. pneumoniae, C. psittaci (avian and feline strains) and C. pecorum) as a cause of CAP. The aim of the present study was to determine the role of C. pneumoniae, C. psittaci and C. pecorum as causes of CAP. A prospective cohort observational study of CAP was conducted at 15 teaching centres in eight Canadian provinces between January 1996-October 1997. Acute (n=539) and convalescent (n=272) serum samples were obtained for determination of antibody titres to C. pneumoniae, C. psittaci, C. pecorum, C. trachomatis, Mycoplasma pneumoniae, Legionella pneumophila serogroups I-VI, Streptococcus pneumoniae and various respiratory viruses. Twelve of 539 (2.2%) patients had acute C. pneumoniae pneumonia and an additional 32 (5.9%) had possible acute infection. C. pneumoniae was the sole pathogen in 16 of 42 (38.1%) of these patients. The most common copathogens were S. pneumoniae, respiratory syncytial virus and influenza virus type A. C. pneumoniae pneumonia patients were older and more likely to show congestive heart failure compared to bacteraemic S. pneumoniae patients. The latter had a lower mean diastolic blood pressure, a higher white blood cell count and a lower arterial carbon dioxide tension. Two patients had antibody titres suggestive of recent infection with the feline strain of C. psittaci. Although numerically Chlamydia pneumoniae is an important cause of community-acquired pneumonia, no distinctive clinical features associated with this pathogen were detected in the present study. Feline Chlamydia psittaci may cause a few cases of community-acquired pneumonia. Avian Chlamydia psittaci should be considered only if there is a compatible epidemiological history. PMID- 12765421 TI - The role of adherence in tuberculosis HIV-positive patients treated in ambulatory regimen. AB - Adherence to antituberculosis treatment, a major determinant of outcome, is of special concern in human immunodeficiency virus (HIV)-positive patients. However, tuberculosis death in HIV-positive patients remains higher than in HIV-negative patients, regardless of adherence. To assess determinants for an unfavourable tuberculosis outcome (defined as no cure or death), and determinants for nonadherence to antituberculosis treatment, 70 HIV-positive patients with tuberculosis referred to an outpatient centre were studied. Patient outcome was compared according to HIV risk factors, other opportunistic diseases, antiretroviral drugs use, current i.v. drugs use, a methadone programme participation, tuberculosis features, treatment characteristics and adherence. Adherent and non-adherent patient characteristics were also compared. An unfavourable outcome occurred in 22.9% of patients and 32.9% were nonadherent with therapy. Nonadherence was the only independent determinant for an unfavourable outcome. Adherence was independently associated with current i.v. drug use, treatment complications and use of methadone. This study confirms that human immunodeficiency virus-infected patients, treated as outpatients, have high rates of nonadherence and that adherence is the strongest determinant for tuberculosis outcome. Independently of the current use of drugs, a methadone programme improves adherence to treatment. PMID- 12765422 TI - Increased tumour necrosis factor-alpha plasma levels during moderate-intensity exercise in COPD patients. AB - Post-training downregulation of muscle tumour necrosis factor (TNF)-alpha messenger ribonucleic acid (mRNA) expression and decrease in cellular TNF-alpha levels have been reported in the elderly. It is hypothesised that chronic obstructive pulmonary disease (COPD) patients may not show these adaptations due to their reduced ability to increase muscle antioxidant capacity with training. Eleven COPD patients (forced expiratory volume in one second 40 +/- 4.4% of the predicted value) and six age-matched controls were studied. Pre- and post training levels of TNF-alpha, soluble TNF receptors (sTNFRs: sTNFR55 and sTNFR75) and interleukin (IL)-6 in plasma at rest and during exercise and vastus lateralis TNF-alpha mRNA were examined. Moderate-intensity constant-work-rate exercise (11 min at 40% of pretraining peak work-rate) increased pretraining plasma TNF-alpha levels in COPD patients (from 17 +/- 3.2 to 23 +/- 2.7 pg x mL(-1); p<0.005) but not in controls (from 19 +/- 4.6 to 19 +/- 3.2 pg x mL(-1)). No changes were observed in sTNFRs or IL-6 levels. After 8 weeks' endurance training, moderate intensity exercise increased plasma TNF-alpha levels similarly to pretraining (from 16 +/- 3 to 21 +/- 4 pg x mL(-1); p<0.01). Pretraining muscle TNF-alpha mRNA expression was significantly higher in COPD patients than in controls (29.3 +/- 13.9 versus 5.0 +/- 1.5 TNF-alpha/18S ribonucleic acid, respectively), but no changes were observed after exercise or training. It is concluded that moderate intensity exercise abnormally increases plasma tumour necrosis factor-alpha levels in chronic obstructive pulmonary disease patients without exercise-induced upregulation of the tumour necrosis factor-alpha gene in skeletal muscle. PMID- 12765423 TI - N-acetylcysteine reduces the risk of re-hospitalisation among patients with chronic obstructive pulmonary disease. AB - The aim of this study was to evaluate the effect of oral N-acetylcysteine in the prevention of re-hospitalisation for chronic obstructive pulmonary disease (COPD) exacerbations. Using the PHARmacoMOrbidity linkage (PHARMO) system the authors included all patients aged > or = 55 yrs who had been dispensed medication, labelled for respiratory indications (anatomical therapeutic chemical (ATC) classification system: R03), between 1986-1998 and who had also been hospitalised for COPD (International Classification of Diseases (ICD)-9: 491, 492, 496) in this time frame. These subjects were subsequently divided into two groups, those who had received N-acetylcysteine following discharge from their first admission between 1986-1998 and those who had not. All the patients were studied starting from their initial discharge, until their first readmission, death or end of data collection period. The maximum follow-up period was 1 yr. A total of 1,219 patients, who were hospitalised for COPD between 1986-1998, were included in this study. After adjustment for disease severity, it was observed that the use of N acetylcysteine was significantly associated with a reduced risk of readmission. The readmission risk was significantly lower in patients with high average daily doses of N-acetylcysteine. In conclusion it was observed that N-acetylcysteine reduces the risk of rehospitalisation for chronic obstructive pulmonary disease by approximately 30% and that this risk reduction is dose-dependent. PMID- 12765424 TI - Monoclonal anti-interleukin-5 treatment suppresses eosinophil but not T-cell functions. AB - Influx of eosinophils in airway mucosa and airway lumen is a hallmark of bronchial asthma. In-vitro data and animal studies indicate that the T-helper (Th) type-2 cell cytokine, interleukin (IL)-5, plays an important role in eosinophil maturation, differentiation, recruitment, and survival. The objective of this study was to determine whether intravenous treatment with monoclonal anti IL-5 would affect the number of peripheral blood eosinophils, their activation status, T-cell activation or the pattern of Th1 and Th2 cytokine production. Over a period of 6 months, 19 asthmatics were investigated in a double-blind, placebo controlled, multicentre study with mepolizumab (SB 240563) anti-IL-5 antibody administered three times. Before each infusion and 12 weeks after the last infusion, peripheral blood leukocytes were examined, qualitative and quantitative distribution of eosinophils and lymphocyte subpopulations, frequencies of IL-2, 3, -4, -5, -10, -13, interferon-gamma-producing CD4 T-cells and serum eosinophil cationic protein (ECP) levels were determined. Treatment with mepolizumab resulted in a marked, rapid and sustained decrease of eosinophil numbers (median values from 300 to 45 per microL) paralleled by decreased levels of serum ECP (median values from 15 to 5 microg x L(-1)). Distribution of T-cell subsets and T cell cytokine production were not altered during antibody treatment. In conclusion, administration of mepolizumab to asthmatic patients markedly reduces peripheral blood eosinophils without altering the distribution and activation status of lymphocytes. PMID- 12765425 TI - Increased immunoreactivity of stromal cell-derived factor-1 and angiogenesis in asthma. AB - Stromal cell-derived factor-1 (SDF-1) acts as a chemoattractant for leukocytes and can induce neovascularisation. To examine the role of SDF-1 in the development of angiogenesis, immunohistochemical studies were performed on bronchial biopsy specimens from asthmatic and control subjects. Bronchial biopsy specimens were obtained from 13 asthmatic and eight control subjects. The number of vessels and the percentage area they occupied were estimated after staining for type-IV collagen. In addition the number of SDF-1-positive cells was determined. There was a significant increase in the number of vessels and the percentage vascularity in the submucosa of asthmatic subjects compared with control subjects. Asthmatic subjects exhibited a greater number of SDF-1-positive cells in the airway mucosa than control subjects. The degree of vascularity was associated with the number of SDF-1-positive cells. Furthermore, the number of SDF-1-positive cells was inversely correlated with airway calibre and airway hyperresponsiveness. Colocalisation studies revealed that endothelial cells, macrophages and T-lymphocytes were the major sources of SDF-1. These findings suggest that increased vascularity of bronchial mucosa in asthmatic subjects is closely related to the expression of stromal cell-derived factor-1 positive cells, which may play a role in remodelling of airways via angiogenesis. PMID- 12765426 TI - Bronchodilator tolerance: the impact of increasing bronchoconstriction. AB - Chronic exposure to beta-agonists causes tolerance to their bronchodilator effects, which is best demonstrated during acute bronchoconstriction. The aim of the present study was to assess whether tolerance becomes more evident with increasing bronchoconstriction, as might occur in acute asthma. In a randomised, double-blind, placebo-controlled, crossover study comprising 15 patients, the treatments were salbutamol 400 microg q.i.d. or placebo given via Diskhaler for 28 days with a 2-week washout between treatments. Patients attended on days 14, 21 and 28. Bronchoconstriction was induced on two of these three occasions to achieve a reduction in the forced expiratory volume in one second (FEV1) of 0 (no methacholine), 15 and 30% (using methacholine) in a randomised order. Immediately after this, salbutamol 100 microg, 100 microg and 200 microg was inhaled at 0, 5, and 10 min. FEV1 was measured over 40 min. Dose/response curves were plotted and values for the area under the curve (AUC)0-40 FEV1 were compared between treatments and by degree of bronchoconstriction. Regular salbutamol resulted in attenuation of the acute response to beta-agonist, which was increasingly evident with greater bronchoconstriction. With a reduction in FEV1 of 0, 15 and 30%, the AUC0-40 FEV1 with salbutamol were 11.2, -14.6 and -35.7% respectively, compared to placebo. There was a linear relationship between the magnitude of bronchoconstriction and the between-treatment differences in AUC0-40 FEV1. Increasing bronchoconstriction conferred greater susceptibility to the effects of bronchodilator tolerance. PMID- 12765427 TI - Protection against methacholine-induced bronchospasm: salbutamol pMDI versus Clickhaler DPI. AB - Passive dry-powder inhalers (DPIs) have been developed as an alternative to pressurised metered-dose inhalers (pMDIs) to improve aerosol delivery on inhalation and eliminate the need for propellants. However, new DPI formulations of generic drugs must be rigorously compared with conventional pMDI therapy. This randomised, double-blind, double-dummy, placebo-controlled, seven-way crossover study evaluated bronchoprotection from methacholine challenge in order to compare a novel salbutamol DPI (Clickhaler) with a reference salbutamol pMDI (Ventolin). Adult asthma patients with airway hyperresponsiveness to methacholine (provocative concentration of methacholine causing a 20% fall in the forced expiratory volume in one second (PC20) <4 mg x mL(-1)) were treated on separate days with 0, 100, 200 or 400 microg of salbutamol via the DPI or pMDI. Methacholine challenge was performed before and after salbutamol treatment and the PC20 ratios analysed by Finney's bioassay to test for therapeutic equivalence of the inhalers. Eighteen patients completed the study and showed significant dose-related responses to salbutamol. The relative potency of DPI:pMDI was 1.29 (90% confidence interval 1.04-1.63). There were no treatment differences in safety (cardiac frequency, blood pressure, adverse events). Methacholine challenge methodology provides a sensitive bioassay and has demonstrated therapeutic equivalence of the salbutamol Clickhaler dry-powder inhaler with the conventional salbutamol pressurised metered-dose inhaler. PMID- 12765428 TI - Social position and mortality from respiratory diseases in males and females. AB - Although social differences in respiratory diseases are considerable, few studies have focused on this disease entity using mortality as an outcome. Does mortality from respiratory disease, including chronic obstructive pulmonary disease (COPD) differ with social position measured by education, income, housing and employment grade? The study population consisted of 26,392 males and females from pooling of two population studies in the Copenhagen area. Data was linked with information from social registers in Statistics Denmark. The relationship between socioeconomic factors and risk of death from respiratory disease and COPD was assessed with an average duration of follow-up of 12 yrs. Education was strongly associated with respiratory mortality in both sexes. The association was stronger in later birth cohorts comparing the highest level of education (>11 yrs) with the lowest (<8 yrs). Although smoking rates were inversely associated with the level of education, the social gradient was not affected by adjustment for smoking. In males, but not in females, there was an additional effect of other indicators of social position, i.e. employment grade (white collar versus blue collar), household income, housing conditions (less than one person per room versus more), and cohabitation (cohabiting versus living alone). Similar results were found for mortality from COPD. The results confirm the existence of a strong social gradient in respiratory mortality and chronic obstructive pulmonary disease, which is independent of smoking and is stronger in males. Social disadvantage is a potentially avoidable cause of death from respiratory disease and further research is needed to explain the excess risk in the socioeconomically disadvantaged. PMID- 12765429 TI - Prevalence and risk factors of asthma and wheezing among US adults: an analysis of the NHANES III data. AB - The prevalence of asthma has been on the increase in the USA and worldwide. To understand the worsening epidemiological trends of asthma, this study analysed the data from the third National Health and Nutrition Examination Survey (NHANES III) to determine the prevalence and risk factors for asthma and wheezing among US adults. This analysis used data from 18,825 US adults aged > or = 20 yrs who had participated in the NHANES III project. After excluding subjects with physician-diagnosed emphysema, a total of 18,393 subjects were included in the final analysis. The prevalence of current asthma (asthma) was 4.5% and the prevalence of wheezing in the previous 12 months (wheezing) was 16.4%. Mexican Americans exhibited the lowest prevalence of asthma when compared with other race/ethnic groups. Multiple logistic regression analysis showed that Mexican Americans were less likely to report asthma when compared to non-Hispanic whites. Low education level, female sex, current and past smoking status, pet ownership, lifetime diagnosis of physician-diagnosed hay fever and obesity were all significantly associated with asthma and/or wheezing. No significant effect of indoor air pollutants, as derived from the use of household heating/cooking appliances, on asthma and wheezing was observed in this study. In conclusion, this study observed racial/ethnic differences in the prevalence of asthma and wheezing and identified several important risk factors that may contribute to development and/or exacerbation of asthma and wheezing. Contrary to earlier reports, the proxy measures of indoor air pollution used in this study were not found to be associated with increased risk of asthma and wheezing. PMID- 12765430 TI - Transient early wheeze is not associated with impaired lung function in 7-yr-old children. AB - The aim of the present study was to analyse determinants of lung function in 7-yr old children with different wheezing patterns (early, persistent and late onset) in a prospective cohort study. The German Multicentre Allergy Study (MAS) followed 1,314 children from birth onwards. Annual assessments included clinical check-ups, a structured interview and repeated measurement of specific immunoglobulins Ig(E) directed against food and inhalant allergens. At the age of 7 yrs, lung function was measured by body plethysmography in 800 children. Episodes of wheezing in the past 12 months ("current wheeze") were strongly associated with reduced lung function at age 7 yrs. Children with wheezing episodes only during the first 3 yrs of life showed a slight impairment in maximal expiratory flow when 50% of the forced vital capacity remains to be exhaled (98.9 +/- 24.2 versus 103.2 +/- 22.8% of the predicted value in children who never wheeze). Separate analysis of determinants of pulmonary function within these subgroups resulted in distinctly different patterns. Determinants of impaired lung function in the group of current wheezers were: time in years since first wheeze, a parental history of atopy, current sensitisation to indoor allergens, elevated cord blood IgE levels and a low ponderal index at birth. In the group of transient early wheezers, frequent lower respiratory tract infections early in life and maternal smoking during pregnancy were significant but weak determinants of impaired lung function. The present results indicate that determinants of pulmonary function in 7-yr-old children differ with respect to different wheezing phenotypes, demanding different therapeutic strategies. Although transient early wheezers were found to have normal-to-subnormal lung function, children with asthmatic symptoms (persistent and late-onset disease) at age 7 yrs already show significant impairment of expiratory flow volumes. PMID- 12765431 TI - Dexamethasone treatment does not inhibit fibroproliferation in chronic lung disease of prematurity. AB - Pulmonary fibrosis results from excessive fibroblast proliferation and increased collagen deposition and occurs in chronic lung disease of prematurity (CLD). Platelet-derived growth factor (PDGF)-BB is mitogenic for fibroblasts and levels are increased in fibrotic lung disorders. Systemic dexamethasone (DEX) treatment improves pulmonary function and reduces inflammation in infants with or at risk of CLD. However, the effect of DEX treatment on fibroblast activity, PDGF-BB and collagen synthesis in the lungs of CLD patients is uncertain. Bronchoalveolar lavage (BAL) fluids, obtained from 15 infants at risk of CLD before and after DEX treatment, were analysed for fibroblast mitogenicity, PDGF-BB, N-terminal propeptide of collagen type III (PIIINP) and interleukin (IL)-1beta levels and inflammatory cell numbers. After DEX treatment, the mitogenic activity of BAL fluid for fibroblasts was not reduced but increased. The change in mitogenicity correlated with a change in BAL fluid PDGF-BB levels. Furthermore, BAL fluid induced fibroblast proliferation was blocked using an inhibitor of the PDGF receptor. DEX treatment did not influence PIIINP levels, but reduced IL-1beta levels and inflammatory cell numbers in BAL fluid. This study suggests that dexamethasone treatment does not reduce fibroblast proliferation despite apparent downregulation of inflammation. The present findings do not support the use of dexamethasone for prevention of the fibrotic response in infants at risk of chronic lung disease of prematurity. PMID- 12765432 TI - Uptake of 18fluorodeoxyglucose in the cystic fibrosis lung: a measure of lung inflammation? AB - Positron emission tomography is a three-dimensional imaging technique that measures physiological effects, including metabolism. 18Fluorodeoxyglucose has been extensively used as a tracer of cellular energy metabolism in the brain and in tumour detection. As neutrophils utilise glucose as an energy source during their respiratory burst, it was hypothesised that 18fluorodeoxyglucose uptake, by these cells, could be interpreted as a measure of neutrophil activation in cystic fibrosis (CF). Ten adult CF patients were given a bolus intravenous injection of 18fluorodeoxyglucose, followed by a 90-min dynamic mid-lung acquisition scan. Right-lung 18fluorodeoxyglucose uptake was assessed using a Patlak plot and values were converted to glucose utilisation. Three clinically inactive pulmonary sarcoidosis patients served as controls. From the 10 CF patients with baseline sputum neutrophils of 14 x 10(6) cells x mL(-1) who were investigated, seven were found to have sputum at a normal or slightly depressed glucose utilisation rate (mean 1.33 micromol x g(-1) x h(-1)) compared with a mean of 2.82 micromol x g( 1) x h(-1) for the sarcoidosis patients. In eight patients, receiving inhaled tobramycin therapy, no change in lung glucose utilisation or sputum neutrophil counts were found. Despite high-sputum neutrophil levels, lung glucose utilisation was not elevated in patients with cystic fibrosis. PMID- 12765433 TI - Pulmonary perfusion quantified by electron-beam computed tomography: effects of hypoxia and inhaled NO. AB - Patients with acute lung injury may benefit from the manipulation of pulmonary blood flow using inhaled nitric oxide (iNO) to optimise ventilation/perfusion matching. Current techniques for studying changes in regional pulmonary perfusion are difficult to apply clinically. This study therefore investigated the potential of electron-beam computed tomography (EBCT) to quantify the effects of hypoxia and iNO on regional pulmonary perfusion in five healthy subjects. Contrast-enhanced sections were obtained sequentially under conditions of normoxia, hypoxia (fractional concentration of oxygen in inspired gas (FI,O2) 0.12) and hypoxia, with iNO (14.8 parts per million (ppm)) administered during inspiration in the supine position. Regions of interest were placed along the nondependent to dependent axis and values for relative perfusion derived. Under normoxic conditions a vertical gradient of perfusion existed, which became less apparent due to increased perfusion in nondependent regions after the induction of hypoxia (FI,O2 0.12). The addition of iNO (FI,O2 0.12 and NO 14.8 ppm) increased perfusion in all regions of the lung section, suggesting redistribution of pulmonary perfusion from other regions of the lung. Absolute values of perfusion were comparable to those documented with existing techniques. The use of a high spatial-resolution technique confirmed the presence of marked perfusion heterogeneity between anatomically close regions of lung. PMID- 12765434 TI - Dehydroepiandrosterone sulphate reduces chronic hypoxic pulmonary hypertension in rats. AB - Pathogenesis of pulmonary hypertension includes vascular smooth muscle cell membrane depolarisation and consequent calcium influx. Usually, calcium-gated potassium channels are activated under such conditions and repolarise the membrane. However, in pulmonary hypertension they are downregulated. The authors hypothesised that pharmacological augmentation of these channels would reduce pulmonary hypertension. Dehydroepiandrosterone sulphate (DHEA-S, 0.1 mg x mL( 1)), a recently characterised activator of calcium-gated potassium channels, was given to rats in drinking water. Pulmonary arterial blood pressure, increased by 4 weeks of hypoxia (from 15 +/- 0.2 to 29.4 +/- 2.5 mmHg), was selectively attenuated in rats treated with DHEA-S for the whole duration of the hypoxic exposure (23.9 +/- 0.9 mmHg) and in rats given DHEA-S only after pulmonary hypertension had fully developed (last 2 weeks of hypoxia; 24.4 +/- 1.4 mmHg). Pulmonary vascular remodelling and right ventricular hypertrophy associated with pulmonary hypertension were also reduced by DHEA-S. Cardiac index and systemic arterial blood pressure did not differ among the groups. The authors conclude that treatment with an activator of calcium-gated potassium channels, dehydroepiandrosterone sulphate, known to be well tolerated by humans, reduces hypoxic pulmonary hypertension in rats. PMID- 12765435 TI - Effects of tobacco smoking on findings in chest computed tomography among asbestos-exposed workers. AB - This study examined the effects of smoking on the findings in chest computed tomography (CT) in 587 asbestos-exposed construction workers (11 females, 576 males, mean age 62 yrs; 18 never-smokers, 406 exsmokers and 163 current smokers). The workers were imaged with spiral CT and high-resolution CT. A total of 13 radiological signs were scored by three radiologists independently. The workers' medical data, smoking habits and occupational exposures were collected at an interview. The effects of smoking status and smoked pack-yrs (0-87.5) on the CT signs were studied using multivariate analysis adjusted for confounding factors. Smoking increased all emphysema signs and contributed to bronchial wall thickening. Smoking was negatively associated with curvilinear and septal lines as well as with parenchymal bands. In persons who had smoked <10 pack-yrs, smoking was positively related to paraseptal emphysema and to bronchial wall thickening and negatively related to septal lines, subpleural nodules and honeycombing. Smoking was related to several abnormal computed tomography signs even among those with relatively small exposure. Computed tomography can detect changes due to smoking at an early stage. PMID- 12765436 TI - Spirometry in young children: should computer-animation programs be used during testing? AB - Currently, computer-animation programs are frequently used to instruct and stimulate young children in performing maximal expiratory flow/volume (MEFV) curves. The reproducibility and maximal performance of MEFV manoeuvres with and without the use of two computer-animation programs (the "candles" and the "balloon" programs) were evaluated. Eighty-eight children, aged 4-8 yrs, were randomly assigned to one of the two animation programs. All children performed two series of at least three technically acceptable curves, one series with the incentive and one without, in random order. With the use of computer-animation programs, a lower proportion of children were able to fulfil international criteria for forced vital capacity (FVC) and forced expiratory volume in one second (FEV1) reproducibility. The use of incentives improved reproducibility and performance of peak expiratory flow (PEF). Performance of FVC decreased significantly in 6-8-yr-old children using the animation programs. Training with a program for a short period of time before the formal lung-function test may be valuable. According to the results, however, the use of these programs during tests under the guidance of an experienced lung-function technician cannot be routinely recommended because of possible deteriorating effects on reproducibility and performance of forced expiratory manoeuvres. PMID- 12765437 TI - To clip or not to clip? Noseclips for spirometry. AB - The use of noseclips for open-circuit spirometry is sporadic, despite guidelines encouraging their use. The authors aimed to evaluate whether noseclips significantly affected measurements of forced vital capacity (FVC) and forced expiratory volume in one second (FEV1) in children attending a tertiary, paediatric respiratory centre. Children attending the asthma and cystic fibrosis (CF) clinics were asked to perform two sets of spirometry, one with and one without noseclips in random order, 20-min apart. Paired data was obtained on 62 patients (32 asthma, 30 CF) with a median age of 11.4 yrs (range 7.2-17.2 yrs). There were no systematic differences in FEV1 or FVC measured with and without noseclips, although seven children (11%) had clinically significant differences in FEV1 of >190 mL. There is no clear advantage to wearing noseclips when performing open-circuit spirometry. Individuals should be assessed to ascertain their optimal technique, which should then be used consistently in clinical practice. Noseclips should probably be retained for research protocols. PMID- 12765438 TI - Diagnosing occupational asthma: how, how much, how far? AB - Diagnosing occupational asthma is still a challenge because it is based on a stepwise approach in which the depth of investigative means may vary depending on resources. The authors herewith review the existing investigative means from the approach of outlining controversies and queries. There is no validated clinical questionnaire for diagnosing occupational asthma. Immunological investigation is limited by the lack of standardised extracts for skin-prick testing and specific immunoglobulin E assessments. Criteria for interpretation of changes in peak expiratory flow rates and bronchial responsiveness to pharmacological agents are still open to discussion. It is worth improving the methodology of specific inhalation challenges, either in the laboratory or in the workplace, to facilitate more extensive use of these tests. Validation of new means that assess airway inflammation, such as exhaled nitric oxide and induced sputum, needs to be performed. There is a need to increase the use of these diagnostic tests because the diagnosis is still too often based on "clinical impression". PMID- 12765439 TI - Varicella pneumonia in adults. AB - Varicella is a common contagious infection in childhood with increasing incidence in adults. Pneumonia, although rare, is the most serious complication that commonly affects adults. Over the last two decades there have been major advances in the understanding of Varicella infections, management and prevention. This review discusses the epidemiology, pathogenesis, pulmonary manifestation, morbidity, long-term clinical consequences and current state of management of Varicella pneumonia in adults. Prevention and other disease-modifying therapy are also discussed. PMID- 12765440 TI - Pulmonary hypertension in chronic obstructive pulmonary disease. AB - Pulmonary hypertension is a common complication of chronic obstructive pulmonary disease (COPD). Its presence is associated with shorter survival and worse clinical evolution. In COPD, pulmonary hypertension tends to be of moderate severity and progresses slowly. However, transitory increases of pulmonary artery pressure may occur during exacerbations, exercise and sleep. Right ventricular function is only mildly impaired with preservation of the cardiac output. Structural and functional changes of pulmonary circulation are apparent at the initial stages of COPD. Recent investigations have shown endothelial dysfunction and changes in the expression of endothelium-derived mediators that regulate vascular tone and cell growth in the pulmonary arteries of patients with mild disease. Some of these changes are also present in smokers with normal lung function. Accordingly, it has been postulated that the initial event in the natural history of pulmonary hypertension in COPD could be the lesion of pulmonary endothelium by cigarette-smoke products. Long-term oxygen administration is the only treatment that slows down the progression of pulmonary hypertension in chronic obstructive pulmonary disease. Nevertheless, with this treatment pulmonary artery pressure rarely returns to normal values and the structural abnormalities of pulmonary vessels remain unaltered. Vasodilators are not recommended on the basis of their minimal clinical efficacy and because they impair pulmonary gas exchange. Recognition of the role of endothelial dysfunction in the physiopathology of pulmonary hypertension in chronic obstructive pulmonary disease opens new perspectives for the treatment of this complication. PMID- 12765441 TI - Recurrent tracheal mass. PMID- 12765442 TI - Efficacy of acupuncture in asthma. PMID- 12765443 TI - Asthma variability. PMID- 12765444 TI - Atrial fibrillation therapy: rate versus rhythm control. PMID- 12765445 TI - Atrial activation analysis by surface P wave and multipolar esophageal recording after cardioversion of persistent atrial fibrillation. AB - We studied atrial activation during sinus rhythm by combining 12-lead ECG and multipolar esophageal recordings in 30 patients after electrical cardioversion of persistent atrial fibrillation. The primary endpoint was to establish a correlation between atrial activation evaluated by the two methods. Total P wave duration and morphology in inferior leads identified three patterns: normal P wave, late-positive P wave, and late-negative P wave. Proximal and distal esophageal recording characterized the longitudinal direction of activation of the posterior left atrium. We distinguished three activation patterns: normal activation when the interatrial conduction time is normal and depolarizes in craniocaudal direction, delayed activation when the interatrial conduction time is prolonged and the craniocaudal activation is maintained, and finally reversed activation when the posterior left atrium depolarizes in a reversed caudocranial direction. Four patients showed a normal P wave and also had a normal esophageal activation. Twelve patients showed a prolonged P wave (associated with delayed esophageal activation in 10 patients and reversed activation in 2 patients); 14 patients had a late-negative P wave (all associated with a reversed esophageal activation). A high correlation existed between each pattern obtained by surface ECG and esophageal recording (P < 0.001) and between surface P wave duration and interatrial conduction time (R2 = 0.64, P < 0.001). Much information concerning atrial activation can be obtained by meticulous analysis of the P wave, particularly its terminal part. Multipolar esophageal recording can be used when surface ECG appears unclear. PMID- 12765446 TI - An adaptive interval-based algorithm for withholding ICD therapy during sinus tachycardia. AB - Avoiding inappropriate ICD therapy during supraventricular tachycardia (SVT) while assuring 100% sensitivity for VT/VF remains a challenge. Inappropriate VT/VF therapy during sinus tachycardia (ST) is particularly distressing to the patient because the full sequence of ICD therapies is often delivered. ST or 1:1 atrial tachycardia (AT) with long PR intervals and ST or AT with atrial oversensing of far-field R waves cause the majority of inappropriate therapy in the Medtronic GEM DR (Model 7271) ICD. The goals of the present effort were to define an adaptive interval-based algorithm for withholding VT/VF therapy in dual chamber ICDs during ST and to compare performance of the adaptive algorithm with that of the original ST withholding algorithm in the GEM DR. The adaptive algorithm uses a combination of 1:1 atrial to ventricular conduction pattern, changes in RR intervals and changes in intrinsic PR intervals to establish evidence for or against the presence of ST. Performances of the adaptive and original ST withholding algorithms were compared on 3 databases collected by implanted GEM DR devices. The first database included 684 spontaneous VT/VF episodes. The second database included 216 spontaneous SVT episodes that received inappropriate VT/VF therapy. These databases included up to 2,000 atrial or ventricular sensed or paced events preceding the spontaneous tachycardias. The third database included 320 spontaneous ST/AT episodes for which therapy was appropriately withheld by the GEM DR. Performance of the adaptive algorithm on the third database was predicted rather than directly computed because of record length limitations. VT/VF therapy was classified as "withheld" if evidence of ST remained high for one algorithm (i.e., at least 7 more beats to VT/VF detection) at the point of VT/VF detection by the other algorithm. For the 684 true VT/VF episodes, the original algorithm withheld VT/VF therapy in 5 episodes and the adaptive algorithm withheld VT/VF therapy in 3 episodes. The 95% confidence interval for the difference in VT/VF sensitivity between the adaptive and original algorithms was [-0.5 to + 1.1%]. Twelve of the 320 ST/AT episodes (3.8%) that were appropriately classified by the original algorithm were predicted to receive inappropriate therapy by the adaptive algorithm. However, relative to the original algorithm, the adaptive algorithm appropriately withheld VT/VF therapy for 76 of 216 true SVT episodes (i.e., incremental specificity of 35.2%). For the specific SVT episodes that were the targets for improvement by the adaptive ST algorithm (ST/AT with long PR intervals and ST/AT with intermittent atrial oversensing of far-field R waves), the adaptive algorithm reduced inappropriate therapy by 63.2%. PMID- 12765447 TI - Myocardial perfusion and adrenergic innervation in patients with RBBB and LAfB: the effect of altering the activation sequence with right ventricular apical pacing. AB - The aim of this study was to investigate myocardial perfusion and adrenergic innervation in patients with intraventricular conduction disturbances and to detect any changes caused by alteration of the ventricular activation sequence as a result of right ventricular apical pacing. We studied 15 patients with right bundle branch block (RBBB) and left anterior fascicular block (LAFB), while 15 healthy individuals served as controls. All patients underwent planar and single photon emission computed tomography (SPECT) myocardial imaging after intravenous infusion of 5mCi 123I-metaiodobenzylguanidine (123I-MIBG) and a SPECT thallium201 myocardial perfusion study before and 3 months after pacemaker implantation. The heart to mediastinum ratio was calculated during the 123I-MIBG study in order to assess the global cardiac sympathetic activity and was significantly smaller in patients than in controls (P < 0.001). Patients with RBBB and LAFB revealed regional adrenergic innervation defects, mostly in the inferior and posterior walls. After a medium-term pacing period, a redistribution of 123I-MIBG uptake was detected, with aggravation of adrenergic innervation defects in the apical and posterior walls and amelioration in septal and anterior walls. Five patients showed perfusion defects that remained unchanged after pacing. Two others displayed mild myocardial perfusion defects that did not exist before pacing. In conclusion, patients with RBBB and LAFB reveal global and regional disturbances of myocardial adrenergic innervation, which shows redistribution as a result of the altered propagation of the ventricular electrical activation. To a smaller degree these patients reveal myocardial perfusion disturbances in which pacing has a limited medium-term effect. PMID- 12765448 TI - Single versus dual chamber pacing in the young: noninvasive comparative evaluation of cardiac function. AB - The advantages of atrial synchrony over asynchronous ventricular pacing remain unclear in the young, chronically right ventricular (RV) - paced patient. This is in contrast to the older patient with inherent diastolic dysfunction who has been shown to benefit from atrial synchrony with dual chamber (DDD,R/VDD), over single chamber rate response (VVI,R) ventricular pacing. The goal of this study was to noninvasively assess cardiac function in a group of young, RV-paced patients before and after establishment of atrial synchrony. Echocardiographic data were retrospectively analyzed from 10 patients with congenital or acquired complete AV block, who were VVI,R paced for 10.2 +/- 2 years (mean age at study 19.2 +/- 8.9 years), and were subsequently converted to DDD,R/VDD pacing (mean age at study 20.7 +/- 9.5 years). Paired t-test analysis of left ventricular (LV) systolic and diastolic function during VVI,R versus DDD,R/VDD pacing did not result in any short-term difference in LV short axis fractional area of change or FAC (53% +/- 7.5% vs 56.8% +/- 8.7%) or mitral maximal velocity (E) normalized to mitral flow velocity time integral (VTI) (5.2/s +/- 1.5 vs 4.4/s +/- 1.5). A decrease in mitral flow E/A ratio was observed after short-term DDD,R/VDD pacing (2.2 +/- 0.5 vs 1.9 +/- 0.3). Atrial synchronous dual chamber pacing in young patients with complete AV block does not lead to any appreciable early change in global LV function over single-site RV pacing. Therefore, early establishment of atrial synchrony in the young asymptomatic VVI,R-paced patient with normal intrinsic ventricular function may not be warranted. PMID- 12765449 TI - AV nodal ablation and pacemaker implantation improves hemodynamic function in atrial fibrillation. AB - In drug refractory and highly symptomatic atrial fibrillation (AF) patients, hemodynamic effects of AV node ablation and pacing therapy (APT) were evaluated. Thirty-eight patients with drug refractory and symptomatic AF, underwent APT in eight centers in Japan. The outcome of this therapy was assessed in terms of quality-of-life, cardiac performance measured by echocardiogram, and plasma ANP and BNP levels before and after APT. Quality-of-life assessed by self administered semi-quantitative questionnaires: WHO QOL 26 (3.0 +/- 0.5 vs 3.4 +/- 0.6, P < 0.01) and the Symptom Checklist: Frequency Scale (1.6 +/- 0.6 vs 0.7 +/- 0.7, P < 0.01) and Severity Scale (1.3 +/- 0.4 vs 0.6 +/- 0.6, P < 0.01), improved significantly 6 months after APT. Ejection fraction (EF) by echocardiogram improved 1 week after APT (59.0% +/- 13.3% vs 63.3% +/- 11.6%, P = 0.02). Plasma ANP levels in the group of ANP > 40 pg/mL at enrollment significantly decreased 1 month later (P = 0.03), and plasma BNP levels in the group of BNP > 20 pg/mL at enrollment significantly decreased 1 month later (P < 0.01). In conclusion, APT has beneficial hemodynamic effects, and plasma BNP levels can predict the most optimal candidates for ablation and pacing therapy. PMID- 12765450 TI - Late improvement in ventricular performance following internal cardioversion for persistent atrial fibrillation: an argument in support of concealed cardiomyopathy. AB - The aim of the study was to evaluate the time course of atrial and ventricular function improvement following internal atrial cardioversion in patients with structural heart disease. Twenty-nine patients with chronic persistent atrial fibrillation (AF) and underlying structural heart disease were followed by serial echocardiograms performed at 1 and 6 hours, 1 day, 1, 2, and 3 weeks, and 1, 2, 3, and 6 months after successful cardioversion. Sinus rhythm was maintained at 6 months in 24 patients. Following cardioversion the time course of left atrial mechanical function (peak A wave, percent A wave filling) differed from that of left ventricular ejection fraction: peak A wave values (cm/s) increased significantly at 1 week (51 +/- 23 vs 35 +/- 15 at 1 hour, P < 0.05), percent A wave filling (%) increased significantly at 2 weeks (34 +/- 12 vs 22 +/- 9 at 1 hour, P < 0.05), whereas left ventricular ejection fraction (%) increased later (at 1 month 60 +/- 14 vs 55 +/- 14 at baseline, P < 0.05 and at 2 months 60 +/- 14 vs 56 +/- 14 at 1 hour, P < 0.05). In conclusion, restoration of sinus rhythm results in an improvement in left ventricular ejection fraction during follow-up, even in patients with structural heart disease without fast ventricular rates at baseline. The dissociation between the time course of atrial and ventricular function improvement suggests that the latter was partly due to regression of a concealed form of cardiomyopathy and/or of a ventricular dysfunction due to chronic AF. PMID- 12765451 TI - High incidence of thrombus formation without impedance rise during radiofrequency ablation using electrode temperature control. AB - The authors hypothesized that during RF ablation, the electrode to tissue interface temperature may significantly exceed electrode temperature in the presence of cooling blood flow and produce thrombus. In 12 anesthetized dogs, the skin over the thigh muscle was incised and raised to form a cradle that was superfused with heparinized canine blood (ACT > 350 s) at 37 degrees C. A 7 Fr, 4 mm or 8-mm ablation electrode containing a thermocouple was held perpendicular to the thigh muscle at 10-g contact weight. Interface temperature was measured at opposite sides of the electrode using tiny optical probes. RF applications (n = 157) were delivered at an electrode temperature of 45 degrees C, 55 degrees C, 65 degrees C, and 75 degrees C for 60 seconds, with or without pulsatile blood flow (150 mL/min). Without blood flow, the interface temperature was similar to the electrode temperature. With blood flow, the interface temperature (side opposite blood flow) was up to 36 degrees C and 57 degrees C higher than the electrode temperature using the 4- and 8-mm electrodes, respectively. After each RF, the cradle was emptied and the electrode and interface were examined. Thrombus developed without impedance rise at an interface temperature as low as 73 degrees C without blood flow and 80 degrees C with blood flow (11/16 RFs at 65 degrees C electrode temperature using 4 mm and 13/13 RFs at an electrode temperature of 55 degrees C using an 8-mm electrode with blood flow). With blood flow, interface temperature markedly exceeded the electrode temperature and the difference was greater with an 8-mm electrode (due to greater electrode cooling). In the presence of blood flow, thrombus occurred without an impedance rise at an electrode temperature as low as 65 degrees C with a 4-mm electrode and 55 degrees C with an 8-mm electrode. PMID- 12765452 TI - Relation of age and sex to atrial electrophysiological properties in patients with no history of atrial fibrillation. AB - Although atrial fibrillation is a common arrhythmia, especially in elderly men, little is known about age related changes in atrial electrophysiological properties or gender differences. The aim of this study was to analyze the effects of aging on vulnerability to atrial fibrillation and assessed gender differences in those age related changes. An electrophysiological study was performed on 73 patients with no history of atrial fibrillation, structural heart disease, or conditions with potential effects on cardiac hemodynamic or electrophysiological function, including 25 women (mean age 49 +/- 18 years; range 12-84 years). The following atrial excitability parameters were assessed: spontaneous or paced (A1) and extrastimulated (A2) atrial electrogram widths, percent maximum atrial fragmentation (A2/A1 x 100), effective refractory period, wavelength index (effective refractory period/A2), and inducibility of atrial fibrillation. There were no significant differences in percent maximum atrial fragmentation (143 +/- 28 vs 142 +/- 35%), effective refractory period (241 +/- 39 vs 238 +/- 50 ms), wavelength index (2.9 +/- 0.8 vs 3.1 +/- 0.9), induction of atrial fibrillation (10 [21%] vs 7 [28%]), or age (50 +/- 17 vs 49 +/- 20 years) between men and women. Age was not statistically different between those patients with and without induction of atrial fibrillation in men (48 +/- 14 vs 50 +/- 18 years) and women (48 +/- 18 vs 49 +/- 21 years). Percent maximum atrial fragmentation and effective refractory period were directly correlated with age in men (r = 0.35, P = 0.01; r = 0.46, P < 0.001, respectively) and women (r = 0.42, P = 0.04; r = 0.45, P = 0.02, respectively), though wavelength index did not correlate with age in men (r = -0.04) or women (r = -0.04) with no history of atrial fibrillation. Considering these findings, the authors conclude that the mechanism triggering atrial fibrillation may be different between older and younger patients with atrial fibrillation, because younger patients who have no marked substrate for atrial fibrillation may need many trigger beats to induce atrial fibrillation. PMID- 12765453 TI - Utility and safety of prolonged temporary transvenous pacing using an active fixation lead: comparison with a conventional lead. AB - Transvenous temporary pacing is associated with a substantial dislocation rate reported to range from 10 to 37%. The aim of the study was to assess the safety and utility of a recently introduced 3.5 Fr temporary pacing lead using active fixation in a consecutive series of 36 patients with prolonged (> or = 48 hours) transvenous temporary pacing (validation group). A group of 36 patients with prolonged transvenous pacing managed with a passive-fixation lead just prior to the introduction of the active-fixation lead served as a control group (reference group). Pacing related adverse events included dislocation, inappropriate pacing (i.e., two-fold or greater increase of initial pacing threshold), local infection, and thrombosis. There were no significant differences in patient characteristics or duration of pacing (5.84 +/- 2.4 days in the reference group vs 5.94 +/- 2.6 days in the validation group). Acute pacing threshold was significantly higher in the validation group as compared to the reference group (1.38 +/- 0.67 V vs. 0.7 +/- 0.21 V, P < 0.01). The dislocation rate was significantly lower in the validation group as compared to the reference group (5.5 vs 33.3%, P < 0.001). There were 11 (31%) pacing related adverse events in the validation group versus 21 (58%) in the reference group (P < 0.01). The vast majority of patients in the validation group (75%) had ambulatory temporary pacing. Thus, transvenous temporary pacing using active fixation is safe and is associated with a low dislocation rate and a reduction in pacing related adverse events. PMID- 12765454 TI - Dose-response relationship for successful internal atrial defibrillation. AB - The dose-response relationship for successful defibrillation has been determined in man for the ventricle but not for the atrium. The purpose of this study was to determine the dose-response relationship for internal atrial defibrillation in humans. Seventy-seven consecutive patients underwent internal atrial defibrillation for acute (n = 14) or chronic AF (n = 63). Shocks were delivered in 40-V increments between electrodes positioned in the coronary sinus and the right atrium until successful conversion or a maximum of 400 V was reached. The shock strength versus success of shock data were subjected to a Kaplan-Meier survival analysis combined with a nonparametric probability analysis to arrive at the dose-response relationship. Using this relationship, comparisons were made between acute and chronic AF and clinical relevant conversion percentages (20, 50, 80 and 95%) were estimated and were compared with the conventional mean threshold. There were significant dose-response relationships in both patients groups (P < 0.05). The Kaplan-Meier analysis comparing patients with chronic and acute AF showed significant differences in their dose-response relationships (P < 0.001). The estimated shock intensity for 95% conversion in patients with acute and chronic AF was 279 V (2.9 J) and 433 V (6.6 J), respectively (P < 0.001). The conventional mean defibrillation threshold in patients with acute (192 +/- 15 V. 1.4 +/- 0.2 J) and chronic AF (343 +/- 8 V, 4.4 +/- 0.2 J) predicted the 60% and 45% chance of successful conversion, respectively. In conclusion, this study demonstrates that single shock conversion data can be used to determine a dose response relationship, which can be used to estimate the shock intensity required for specific successful atrial defibrillation efficacy and to compare different clinical factors that affect defibrillation efficacy. PMID- 12765456 TI - Significance of the morphological patterns of electrograms recorded during ventricular fibrillation: an experimental study. AB - Mapping techniques are used to study the significance of the morphological patterns of the electrograms (EGMs) obtained during VF in an experimental model. In 24 isolated rabbit heart preparations recordings were made of activation during VF using a multiple electrode (121 unipolar electrodes) positioned on the lateral wall of the left ventricle. Three types of activation maps were selected: (A) with functional block of an activation front; (B) with epicardial breakthrough; and (C) with a single broad wavefront without block lines. The EGMs were classified as negative (Q), positive-negative with a predominance of the negative (rS) or positive wave (Rs), and positive (R). In 60 type A maps the morphology in the zone limiting the block line corresponded to an R wave in 55 (92%) cases and to Rs in 5 (8%) cases. In 67 type B maps, the EGM in the earliest activation zone most often showed Q wave morphology (48 [72%] cases), followed by rS (18 [27%] cases), and Rs morphology (1 [1%] case); in no case was R wave morphology seen. Finally, in 78 type C maps the morphology corresponded to a Q wave in 15 (19%) cases, rS in 38 (49%), Rs in 24 (31%), and R in a 1 (1%) case. The differences between the three types of maps were significant (P < 0.0001). Q wave EGM sensitivity for indicating the existence of an epicardial breakthrough pattern was 72%, with a specificity of 89%, and positive and negative predictive values of 76% and 87%, respectively. R wave EGM sensitivity for indicating the existence of conduction block was 92%, with a specificity of 99%, and positive and negative predictive values of 98% and 97%, respectively. R wave morphology is highly sensitive and specific for indicating conduction block. EGM recordings with initial positivity predominance are infrequent in the earliest activation zones of epicardial breakthrough during VF. The recording of the EGM with Q wave morphology indicates centrifugal activation from the explored zone. PMID- 12765455 TI - Effects of sulfonylurea hypoglycemic agents and adenosine triphosphate dependent potassium channel antagonists on ventricular arrhythmias in patients with decompensated heart failure. AB - Hypoglycemic sulfonylureas block cardiac ATP-sensitive potassium channels (K(ATP)). The opening of these channels in cardiomyocytes can induce arrhythmias. In animal studies, sulfonylureas exert an antiarrhythmic effect on the ischemic myocardium, but data on human arrhythmic events are lacking. The study population included 207 patients (age 61 +/- 14 years) admitted for decompensated CHF. The severity of ventricular arrhythmias was assessed by 24-hour Holter monitoring. None of the patients were on parenteral vasoactive therapy or antiarrhythmics during Holter recording. Diabetic patients comprised 48% of the study population, and 34% of diabetic patients were prescribed sulfonylureas. The mean hourly ventricular pairs (3.6 +/- 0.5 vs 1.8 +/- 0.3, P = 0.03), the mean hourly repetitive ventricular beats (5.7 +/- 1.0 vs 2.6 +/- 0.1, P = 0.03), and the frequency of ventricular tachycardia episodes per 24 hours (4.7 +/- 0.8 vs 2.2 +/ 0.4, P = 0.03) were significantly lower in patients with diabetes who were receiving sulfonylureas compared with nondiabetics. No significant difference occurred between patients with diabetes who were not receiving sulfonylureas and nondiabetic patients. Multivariate regression revealed a negative independent relationship between sulfonylurea therapy and hourly ventricular pairs (P = 0.03), the mean hourly repetitive ventricular beats (P = 0.03), and ventricular tachycardia episodes (P = 0.04). In a multiple logistic regression, sulfonylurea therapy was a negative predictor of repetitive ventricular beats (P = 0.01, adjusted OR, 0.31; 95% CI, 0.12-0.78). Concomitant sulfonylurea therapy may reduce the occurrence of complex ventricular ectopy in the setting of severe CHF. These results suggest that cardiac K(ATP) channel activation may be involved in the genesis of ventricular arrhythmias in CHF. PMID- 12765457 TI - Pacemaker mode selection: the evidence from randomized trials. AB - The evidence base for pacing, specifically with regards to outcome-based randomized trials, is only beginning to emerge. At present, the guidelines for pacing in sinus node dysfunction (SND), atrioventricular block (AVB), and vasovagal syncope are largely based on observational, not randomized studies. The findings from observational studies that physiological pacing was associated with reduced mortality, fewer strokes, less heart failure, and less AF when compared with ventricular pacing, were not uniformly supported by the early randomized trials of a relatively small sample size. Thus, it has become increasingly clear that large scale randomized trials are necessary to measure reliably the benefit, if any, of progressively more expensive and complex pacemakers. To provide reliable answers to these important questions, three large multicenter randomized trials in Canada, the United Kingdom, and the United States have been designed and conducted. The present review analyzed the results of completed randomized trials on pacemaker mode selection. To date, > 6,000 patients requiring permanent pacing to prevent bradycardia have been randomized; among these, dual chamber pacing did not prevent stroke or improve survival when compared with ventricular pacing. However, dual chamber pacing led to a moderate reduction of incident and chronic AF, reduced symptoms of heart failure in patients with SND, prevented pacemaker syndrome, and modestly improved quality-of-life. Further, a 5-10% reduction in mortality by atrial-based pacing cannot be excluded based on the results of the analyzed trials. The availability of data from ongoing randomized trials and their meta analysis should complete the totality of evidence during the next several years. PMID- 12765458 TI - "My dear child". PMID- 12765459 TI - Radiofrequency catheter ablation of junctional ectopic tachycardia with preservation of atrioventricular conduction. AB - Junctional ectopic tachycardia is a relatively rare disorder, frequently refractory to drug therapy, and with a poor prognosis in childhood. This report describes a successful radiofrequency catheter ablation of the focus of this arrhythmia in a 9-year-old girl with preservation of normal atrioventricular conduction, using precise catheter navigation with the LocaLisa system and carefully titrated RF delivery. PMID- 12765460 TI - Electrocardiographic changes due to pyridoxine deficiency. AB - A young woman presented with marked alterations in the ECG without cardiological symptoms or evidence of structural heart disease after further evaluation. There was evidence of vitamin deficiency and the ECG normalized after 10 days of treatment with vitamins. Similar alterations have been described in several experimental studies with rats, but this is the first case reported in humans. PMID- 12765461 TI - Electrical interference from an abdominal muscle stimulator unit on an implantable cardioverter defibrillator: report of two consecutive cases. AB - Two consecutive cases are presented of patients with ICDs in whom the use of commercial units for muscle stimulation for abdominal training caused interference with the device, mimicked cardiac signals, and resulted in inappropriate treatment shock delivery. PMID- 12765462 TI - Active-can implantable cardioverter defibrillator placement from a femoral approach. AB - This report describes a case of an active-can ICD placed in the thigh. A 74-year old man on chronic renal dialysis had no venous access from cephalic, subclavian, or jugular approaches. Using long active-fixation leads the device was placed from a femoral approach with good sensing, pacing, and defibrillation parameters. PMID- 12765463 TI - Correct a statement in the article by Hauser et al. Long-term structural failure of coaxial polyurethane implantable cardioverter defibrillator leads. PMID- 12765464 TI - Role of immune cells in pregnancy. AB - The human uterus is generally considered to be an immunologically privilege site that isolates the implanted allogeneic embryo from an aggressive maternal immune response. Maternal tolerance of the fetal allograft could be the result of the integration of numerous mechanisms promoted by decidual natural killer (NK) cells, macrophages and T cells. In this review, we outline the possible role of all these immune cells on the maintenance of pregnancy, focusing on the role of the T cells. PMID- 12765465 TI - Microchimerism in human health and disease. AB - During pregnancy some cells traffic between the fetus and the mother. Recent investigative work indicates a low level of fetal cells commonly persists in the maternal circulation for years, oreven indefinitely, after pregnancy has been completed. The term microchimerism refers to one individual harboring DNA or cells at a low level that derive from another individual. Chronic graft-versus host disease (cGvHD) shares similarities with some autoimmune diseases and is an iatrogenic form of chimerism, occurring as a complication of hematopoietic stem cell transplantation. The HLA genes of the donor and the host are known to be of central importance to the development of cGvHD. When also considered in light of the female predilection to autoimmunity, these series of observations led to the hypothesis that microchimerism and HLA genes of host and non-host cells are involved in some autoimmune diseases. The hypothesis can also apply to men, children, and women who have not been pregnant because there are other sources of microchimerism. Persistent microchimerism can follow a blood transfusion, or can occur from transfer between twins in utereo. Additionally, maternal cells have recently been found to persist in her immune competent progeny. A number of studies have investigated a potential role of microchimerism in human diseases including systemic sclerosis (SSc), primary biliary cirrhosis (PBC), Sjogren's syndrome, polymorphic eruption of pregnancy, myositis, and thyroid disease. While some studies lend support to the concept that microchimerism is involved in the pathogenesis of selected autoimmune diseases, studies also indicate microchimerism is not uncommon in other human conditions and in healthy individuals. PMID- 12765466 TI - Placental trophoblast and endothelial cells as target of maternal immune response. AB - Pregnancy is a unique physiologic condition that guarantees the survival of the semiallogenic embryo during the long period of gestation. The placenta plays a key role in the maintenance of local tolerance and allows the mother to accept the embryo until completion of pregnancy. The complex process of tolerance accompanying the survival of the foetus is controlled at the embryo-maternal interface by factors deriving from decidualized endometrium and from the trophoblast itself. Trophoblasts develop various strategies to evade the damaging attack by the maternal immune response including expression of non-classical MHC class I antigens and of complement regulatory proteins. Also, cytokines released at the feto-maternal interface play an important role in regulating embryo survival controlling not only the maternal immune response but also angiogenesis and vascular remodelling. The delicate equilibrium established between the mother and the foetus can be compromised in pathological condition of pregnancy as a result of humoral and/or cellular response of the mother against trophoblast antigens leading to spontaneous miscarriage. Cytotoxic cells and antibodies to trophoblast and endothelial cells are frequently found in patients with recurrent spontaneous abortion. This review article focuses on the delicate equilibrium established at the feto-maternal interface during pregnancy examining the various strategies devised by the embryo to evade the maternal immune attack, and the pathological conditions in which this equilibrium is compromised leading to serious complications of pregnancy. PMID- 12765467 TI - The role of complement in pregnancy and fetal loss. AB - In the United States, between 1 and 3% of women suffer recurrent miscarriages; 50 70% of all conceptions fail. Although in the majority of affected women the cause of recurrent miscarriages is unknown, an immune mechanism involving the inappropriate and subsequently injurious recognition of the conceptus by the mother's immune system has been proposed. Murine models have recently been developed that are relevant to this issue. We and others have identified a novel role for complement as an early effector in the pathway leading to pregnancy loss associated with placental inflammation. Indeed, it appears that inhibition of complement activation is an absolute requirement for normal pregnancy, and that in the antiphosphospholid syndrome overwhelming activation of complement triggered by antibodies (Ab) deposited in placenta leads to fetal injury. Identification of complement activation as a mediator of pregnancy loss and definition of the complement components necessary to trigger such injury is likely to lead to a better understanding of its pathogenesis and to new and improved treatments. PMID- 12765468 TI - Pregnancy complications of the antiphospholipid syndrome. AB - Starting from their first description, antiphospholipid antibodies (aPL) were associated with repeated miscarriages and fetal losses. Other complications of pregnancy like preterm birth,with pre-eclampsia or severe placental insufficiency were also frequently reported and are included in the current classification criteria of the antiphospholipid syndrome (APS). The titre, the isotype of the antibodies or their antigen specificity may be important in the risk level determination. Some of the difference in the reported results can be explained by the poor standardization achieved in aPL testing or by the not univocal classification of pregnancy complications. The pathogenesis of pregnancy failures is linked to the thrombophilic effect of aPL but also to different mechanisms including a direct effect of antibodies on the throphoblast differentiation and invasion. The study of experimental animal models provided sound evidence of the pathogenic role of aPL both in lupus prone and naive mice. The definition of APS as a condition linked to high obstetric risk and the application of an effective therapy have completely changed the prognosis of pregnancy in these patients. In fact, despite the high number of complications and preterm delivery, today a successful outcome can be achieved in the large majority of the cases. PMID- 12765469 TI - Lupus pregnancy. AB - Pregnancy markedly alters a normal woman's physiology and immune response mechanisms. The effect of pregnancy changes on the course of systemic lupus erythematosus (SLE) remains, however, speculative. Pregnant lupus patients are susceptible to pre-eclampsia, especially if they suffer lupus nephritis, and to steroid-induced hypertension and hyperglycemia. Fetuses are susceptible to placental insufficiency if antiphospholipid antibody or other procoagulant states are present, and to neonatal lupus if anti-Ro/La antibodies are present. Artificial reproductive technologies ("in vitro fertilization") can be safely used in SLE patients. Study of the physiology of pregnancy, for instance complement kinetics, may inform our understanding of SLE, and vice versa. PMID- 12765470 TI - Neonatal lupus: review of proposed pathogenesis and clinical data from the US based Research Registry for Neonatal Lupus. AB - Congenital heart block (CHB), a life-threatening manifestation of neonatal lupus, offers aunique opportunity to study the effect or arm of immunity and define the pathogenicity of an autoantibody in mediating tissue injury. This review focuses on our recent in vitro model which supports a cascade from antibody insult to unchecked fibrosis. In brief, it is proposed that the fetal cardiac myocyte undergoes apoptosis which facilitates transfer of intracellular Ro and La antigens to the surface where they are bound by circulating maternal autoantibodies (anti-SSA/Ro-SSB/La antibodies). Scavenging macrophages phagocytose these inadvertently "opsonized" cardiocytes, leading to the secretion of pro-inflammatory and pro-fibrotic cytokines, the latter of which transdifferentiate fibroblasts into myofibroblasts and thereby promote scarring. Immunohistologic study of a heart from a neonate dying of CHB supports this model in that macrophages and myofibroblasts were demonstrated. To facilitate both basic and clinical research, a Research Registry for Neonatal Lupus was established in 1994 by the U.S. National Institute of Arthritis, Musculoskeletal and Skin Diseases. Maternal and fetal outcomes are addressed as well as recurrence rates. Laboratory evaluation and management decisions during pregnancy are provided. PMID- 12765471 TI - Immunosuppressive drug use in pregnancy. AB - Ideally, immunosuppressive drugs would not be necessary in pregnancy. However, in connective tissue disease (especially systemic lupus erythematosus, SLE) vasculitis, and sometimes antiphospholipid antibody syndrome, their use is necessary both to protect the health of the mother and to insure the success of the pregnancy. The more commonly used drugs will be reviewed, with an emphasis on human data, when available. Methotrexate and leflunamide will not be considered, for they should never be used in pregnancy. PMID- 12765472 TI - Frequency and significance of antibodies to soluble liver antigen/liver pancreas in variant autoimmune hepatitis. AB - BACKGROUND: Antibodies to soluble liver antigen/liver pancreas are highly specific markers of autoimmune hepatitis. AIMS: Determine the frequency and clinical significance of these antibodies in the variant syndromes. METHODS: Antibodies to soluble liver antigen/liver pancreas were determined in 28 patients with variant forms, including 10 with cryptogenic chronic hepatitis and 18 with cholestatic variants. One hundred and seventy-two patients with classical autoimmune hepatitis were similarly tested. RESULTS: Seven of the 28 patients with variant forms had the antibodies, and this frequency was not statistically different than that in classical disease (25 vs. 12%, p = 0.08). Antibodies were most common in patients with cryptogenic chronic hepatitis (40%). Seropositive patients were indistinguishable from seronegative patients with variant forms, and they responded as well to corticosteroid therapy as patients with autoimmune hepatitis. Relapse after corticosteroid withdrawal invariably occurred in the seropositive patients whether with variant or classical disease, and HLA DR3 was more common in the seropositive patients with variant forms than in normal subjects (60 vs. 15%, p = 0.03). CONCLUSIONS: Antibodies to soluble liver antigen/liver pancreas occur commonly in the variant forms of autoimmune hepatitis and identify patients that closely resemble classical disease. Seropositivity is associated with relapse after corticosteroid withdrawal and HLA DR3. The antibodies may be surrogate markers of a genetic propensity to relapse that is independent of clinical phenotype. PMID- 12765473 TI - Anti-soluble liver antigen/liver-pancreas (SLA/LP) antibodies in pediatric patients with autoimmune hepatitis. AB - Antibodies against soluble liver antigen/liver-pancreas (SLA/LP) have been associated with severe autoimmune hepatitis (AIH) and poor outcome, but most of these reports have focused on adult patients. The aim of this study was to assess the prevalence and clinical significance of anti-SLA/LP antibodies in a pediatric population with AIH. We developed a quantitative enzyme-linked immunoassay (ELISA), a Western blot (WB) and an immunoprecipitation assay (IPA) based on recombinant cDNA from activated Jurkat cells. The specificity of these tests was validated by testing 200 serum samples from healthy subjects, and from patients with liver and non-liver diseases. Anti-SLA/LP antibodies were found in patients with type 1 and type 2 AIH. The prevalence of these antibodies in patients with type 1 AIH was: 42% when tested by ELISA, 15% by WB and 50% by IPA. In patients with type 2 AIH, the prevalence rates were 42% by ELISA, 18% by WB and 44% by IPA. The mean titer values for anti-SLA/LP antibodies was significantly higher in type 2 AIH (1:1,300 +/- 339) than in type 1 AIH (1:600 +/- 71; p < 0.0001) and closely associated with higher titers of anti-liver kidney microsome type 1 (LKM1) and anti-liver cytosol type 1 (LC1) antibodies in sera. The presence of anti-SLA/LP showed a significant female preponderance in type 1 and 2 AIH patients (p = 0.0003 and p = 0.003, respectively), and was significantly correlated with a lower age at diagnosis (p = 0.05) in type 1 AIH patients. In conclusion, anti-SLA/LP antibodies in pediatric patients are associated with both type 1 and 2 AIH. PMID- 12765474 TI - Prevalence of autoantibodies to the p53 protein in autoimmune hepatitis. AB - The target antigens of anti-nuclear autoantibodies in autoimmune hepatitis (AIH) are poorly characterised. Since antibodies to the p53 nuclear protein have been reported in various autoimmune diseases, we have assessed the prevalence of these antibodies in patients with AIH (n = 45), primary biliary cirrhosis (n = 60), hepatitis B (n = 22), hepatitis C (n = 55), and in a control group of subjects with various non-liver diseases (n = 56). A significant proportion of patients with AIH (31%) had elevated levels of autoantibodies to the p53 protein. In contrast, the prevalence of these antibodies in primary biliary cirrhosis (8%) and viral hepatitis (6%) was similar to that in the control group (4%). The clinical features of the anti-p53 seropositive AIH patients were similar to those of the seronegative ones. Thus, the prevalence of p53 autoantibodies in AIH is higher than in other forms of chronic hepatitis, and may be useful in differential diagnosis. PMID- 12765475 TI - Smooth muscle antibodies and type 1 autoimmune hepatitis. AB - BACKGROUND/AIM: Smooth muscle antibodies (SMA) characterize type 1 autoimmune hepatitis. Our aim was to evaluate sensitivity and specificity of different immunofluorescence substrates for the detection of SMA. METHODS: Sera from 55 patients with type 1 AIH 20 with primary biliary cirrhosis, 20 with HCV-related chronic hepatitis and 25 blood donors were studied for SMA and anti microfilaments reactivity by immunofluorescence on rat tissue sections, cultured fibroblasts and commercially available HEp-2 cells (collectively revealing the so called anti-actin pattern), and for the XR1 system by counterimmunoelectrophoresis. SMA was classified on the basis of its immunofluorescence pattern (V--vessels, G--glomerular, T--tubular). As further control group, we studied 26 patients with a diagnosis other than AIH, selected on the basis of a SMA-non-T/XR1 positivity. RESULTS: In patients with AIH the SMA T pattern on rodent tissue, and anti-MF on fibroblasts and on HEp-2 cells were present in 80, 82 and 80%, respectively. Five out of 11 SMA-non T positive AIH patients were anti-MF positive. None of the pathological and healthy controls was positive for SMA-T or anti-MF reactivity. XR1 system was present in 84% of AIH patients and in 5% of pathological controls (p = 0.01). Two out of 26 SMA-non T/XR1 positive sera were positive for anti-MF by fibroblasts and HEp-2 cells. A significant correlation was found between SMA-T pattern and anti-MF reactivity; no correlation was found between XR1 system and SMA-T pattern or anti-MF reactivity. CONCLUSIONS: SMA-T pattern is highly sensitive and specific first diagnostic test for type 1 AIH; anti-MF can be used as additional tool for the diagnosis, particularly when, despite the absence of the SMA-T pattern, AIH is strongly suspected. PMID- 12765476 TI - A major CYP2D6 autoepitope in autoimmune hepatitis type 2 and chronic hepatitis C is a three-dimensional structure homologous to other cytochrome P450 autoantigens. AB - Liver-kidney microsomal antibodies type 1 (LKM) are a diagnostic marker for autoimmune hepatitis type 2 (AIH-2), however, LKM autoantibodies are also detected in a small percentage of patients with chronic hepatitis C. The major target of LKM antibodies as evidenced by indirect immunofluorescence is cytochrome P4502D6 (CYP2D6). Anti-CYP2D6 titers of 62 LKM positive sera, 196 sera of patients with hepatic and rheumatic diseases and 33 sera of healthy blood donors (BD) were determined by an in vitro transcription/in vitro translation assay (ITT). Twenty five out of 26 AIH-2 sera and 33/36 LKM positive hepatitis C virus (HCV) sera were anti-CYP2D6 positive by ITT and antibody titers were similar in both patient groups. Epitope mapping experiments were performed by a series of truncated CYP2D6 proteins and by single epitopes of 257-269, 321-351, 373-389 and 410-419 amino acid (aa) expressed as DHFR-fusion proteins in Escherichia coli. The major linear epitope consists of 257-269 aa. This epitope is recognized with a significantly higher prevalence (64%) in AIH-2 than in LKM sera from patients with chronic hepatitis C (24%) (p < 0.001). None of the other autoepitopes showed significant differences in the prevalence of recognition by sera from both patient groups. Minor binding sites consisted of 321-351 aa, which was recognized by less than 20% of LKM sera and in the C-terminal region of 350 494 aa, which was recognized by less than 5% of LKM sera Our study revealed an epitope of 321-379 an on CYP2D6, which was shown to be conformation dependent. It was recognized by the vast majority of LKM sera, specifically by 76% of sera from HCV positive LKM patients and also by 76% of sera from patients with AIH-2. This epitope is homologous to three-dimensional epitopes detected by autoantibodies directed against hepatic cytochromes P450s in drug induced hepatitis and to an autoepitope on CYP21B associated with adrenal failure. PMID- 12765477 TI - Organ and non-organ specific autoantibody titres and IgG levels as markers of disease activity: a longitudinal study in childhood autoimmune liver disease. AB - No longitudinal study has investigated whether autoantibody titres and serum IgG levels correlate with disease activity in autoimmune liver disease. To determine this, we investigated prospectively 19 patients on 254 occasions between 10 months to 5 years from diagnosis. Nine had anti-nuclear and/or anti-smooth muscle antibody (ANA/SMA) positive autoimmune hepatitis (type 1 AIH), 5 liver kidney microsomal type 1 (LKM-1) positive AIH (type 2 AIH) and 5 ANA/SMA positive autoimmune sclerosing cholangitis (ASC). Correlation between IgG levels, titres of ANA, SMA and LKM-1 and levels of the organ specific autoantibodies anti-liver specific protein (anti-LSP), and anti-asialoglycoprotein receptor (anti-ASGPR) with biochemical evidence of disease activity, as measured by serum aspartate amino transferase (AST) levels, was sought during the course of the disease. AST levels correlated with levels of anti-LSP, anti-ASGPR and IgG in type 1 and 2 AIH, but not in ASC. Positive correlation with AST was also observed for LKM-1 titres in type 2 AIH and for SMA titres in type 1 AIH, but not in ASC. In both AIH and ASC, AST levels correlated with the T cell-dependent immune responses anti rubella IgG and anti tetanus toxoid IgG, but not with the T cell-independent IgG2 response to pneumococcal capsular polysaccaride. Our results indicate that measurement of organ and non-organ specific autoantibodies and IgG levels may be used to monitor disease activity in AIH. PMID- 12765478 TI - Elevated expression of tyrosine kinase DDR2 in primary biliary cirrhosis. AB - BACKGROUND: Primary biliary cirrhosis (PBC) is characterized by chronic progressive destruction of small intrahepatic bile ducts with portal inflammation and cholestasis, leading to fibrosis. METHODS: We utilized a novel restriction analysis system to profile the expression of tyrosine kinases (TKs). This methodology targets a conserved sequence present in the majority of human TKs, and exploits the known restriction map of the TK cDNA sequences. We isolated mRNA from biliary epithelial cell (BEC)-enriched cell fractions, amplified the TK transcripts using degenerative primers, and identified specific TKs by restriction enzyme digest analysis and then performed in situ hybridization. RESULTS: BEC-enriched samples from PBC livers displayed marked expression of discoidin domain receptor-2 (DDR2), whereas, non-diseased livers showed no detectable DDR2. Furthermore, in situ hybridization of PBC livers revealed that DDR2 is expressed in the small bile duct epithelial regions as well as in fibroblasts/stromal cells of fibrotic regions. A similar pattern was observed in livers of primary sclerosing cholangitis (PSC), although the amount of small ducts that were positively stained was lower than in PBC. Furthermore, cirrhotic livers of patients with other diseases, including alcoholic liver disease and chronic hepatitis C, DDR2 transcripts were noted only within fibrotic lesions and the degree of intensity was much lower than in PBC and PSC. CONCLUSIONS: DDR2, a TK that is stimulated by fibrillar collagens that accumulate in cirrhotic livers, is present at elevated levels in the small bile ducts of PBC patients. DDR2 is part of a positive feedback loop in which its enhanced expression leads to enhanced deposition of fibrillar collagens (types I andIII). These fibrillar collagens can also provide binding sites for immune mediators, such as cytokines and chemokines. Therefore, unusually high DDR2 expression in the bile ducts of PBC patients could contribute to duct injury by altering local cytokine levels and thereby increasing immune-mediated damage. PMID- 12765479 TI - Association of single nucleotide polymorphisms of the interleukin-10 promoter gene and susceptibility to primary biliary cirrhosis: immunogenetic differences in Italian and Japanese patients. AB - Several lines of data suggest that genetic factors play an important role in the onset and/or progression of primary biliary cirrhosis (PBC). Since PBC is an autoimmune disease, it is reasoned to assume that genes encoding cytokines may confer susceptibility to disease. Amongst these factors, interleukin-10 (IL-10) has received significant attention. The promoter region of IL-10 gene has three single nucleotide polymorphisms (SNPs) at positions -1082, -819 and -592. To elucidate the association of the three SNPs of IL-10 promoter region with susceptibility of PBC in two different genetic populations, 159 unrelated patients with PBC (94 Italian and 65 Japanese) and 143 local controls (72 Italian and 71 Japanese) were enrolled. SNPs were determined using allele-specific PCR/RFLP. In Italian PBC patients, the frequency of homozygosity for G/G at position -1082 was significantly higher than that of local controls (p < 0.041, OR = 2.44, 95% C.I.; 1.02-5.86). The frequencies of haplotype GCC in PBC patients, possibly linked to higher IL-10 production, were also significant higher than local controls (p < 0.033). However, in Japanese population, there were no significant differences in the three SNPs and haplotypes between PBC patients and controls. Excessive production of IL-10 may play an important role in some populations in modulating the onset of PBC. Further, immunogenetic studies of PBC should take into account ethnic and geographic variations; this makes such studies in heterogeneous population, like the USA, more difficult. PMID- 12765480 TI - Oral tolerisation to pyruvate dehydrogenase complex as a potential therapy for primary biliary cirrhosis. AB - Primary biliary cirrhosis (PBC) is characterised by immune-mediated damage to the intra-hepatic biliary epithelial cells (BEC). Immuno-modulatory/suppressive therapy represents, therefore, alogical approach to treatment in this disease. Conventional immuno-suppressive and immuno-modulatory agents suffer from the breadth of their action and/or excessive side effects. Autoreactive responses to pyruvate dehydrogenase complex (PDC) have been extensively characterised in PBC, and implicated in target cell damage. The aim of the current study was to study the potential efficacy of an antigen specific approach (oral tolerisation with autoantigen) to modulation of anti-PDC immune responses characteristic of PBC, utilising a mouse model of PDC immuno-reactivity. Groups of SJL/J mice were orally dosed with PDC alone, dosed with carrier only (saline) but systemically sensitised with PDC in adjuvant, or orally dosed with PDC at high (5 mg) or low (0.01 mg) dose and systemically sensitised with PDC. Oral dosing with PDC in isolation had no adverse effects on the animals and did not prime anti-PDC responses at doses below 1 mg. Pre-dosing with PDC at both high and low doses was effective at skewing the phenotype of the T-cell response to PDC induced by subsequent sensitisation away from the disease associated Th-1 phenotype (IL-2 and IFN-gamma secreting) and towards a theoretically protective Th-2 phenotype (IL-4 secreting) in a majority of, but not all, animals. No augmentation of Th-1 response was seen in any animal. Although the effects on liver histology remain to established oral tolerisation with PDC holds promise as a novel, antigen specific approach to therapy in PBC. PMID- 12765481 TI - Neonatal liver disease associated with placental transfer of anti-mitochondrial antibodies. AB - BACKGROUND: Anti-mitochondrial antibody is the diagnostic hallmark of primary biliary cirrhosis. Its role in the aetiology of primary biliary cirrhosis is controversial. METHODS: Two cases of neonatal hepatitis seropositive for anti mitochondrial antibody are described. Anti-mitochondrial antibody Ig isotype and epitopic specificity were investigated by immunofluorescence and enzyme immunoassays. RESULTS: In both infants anti-mitochondrial antibody was of the G class, mainly G1 and G3 subclasses, and reacted with two synthetic peptides reproducing major M2 epitopicregions: innerlipoyl domain pyruvate dehydrogenase complex (PDC)-E2(162-176) and PDC-E3 binding protein (PDC-E3BP)86-100. One infant also reacted with outer lipoyl domain PDC-E2(35-49), and 2-oxoglutarate dehydrogenase complex (OGDC)-E2(99-113). An identical pattern of reactivity was present in their mothers, indicating the maternal origin of the antibodies. Anti mitochondrial antibody disappeared in the infants with the disappearance of the liver pathology. CONCLUSIONS: The simultaneous disappearance of hepatitis and anti-mitochondrial antibody in the infants suggests a possible causal link between the two. PMID- 12765482 TI - Levels of BAFF in serum in primary biliary cirrhosis and autoimmune diabetes. AB - Levels of the B-cell activating cytokine BAFF are increased in serum in various autoimmune disease, and particularly Sjogren's syndrome in which there is evident B-lymphocyte proliferation. Studies in two autoimmune disease in which B-cell proliferation is less evident, primary biliary cirrhosis (PBC), and adult-onset Type 1 diabetes, showed serum levels of BAFF to be mostly in the normal range. A single raised level among eight sera tested in one patient studied with autoimmune hepatitis (AH) coincided with a relapse of the disease. Increased levels of BAFF in human sera, indexing a potent antigenic drive on B-cell production and survival in some autoimmune diseases, may mark only particular stages in the evolution of such diseases. PMID- 12765483 TI - Evaluation of the role of MHC class II alleles, haplotypes and selected amino acid sequences in primary sclerosing cholangitis. AB - BACKGROUND AND AIMS: Genetic susceptibility to primary sclerosing cholangitis is associated with several different HLA haplotypes, though a single "shared" susceptibility allele has yet to be identified. Most recently, attention has focussed on the MICA alleles in close proximity to the HLA class I, B locus. However, although there are strong associations with MICA*008, implicating this or a closely linked allele as major risk factors, this explanation alone does not account for all of the MHC-encoded susceptibility and resistance to PSC. The present study re-examines HLA class II associations in a large single centre series of well-characterised PSC patients. The specific aims of the study were to test existing associations and to develop hypotheses which together may account for all, or the majority, of the MHC-encoded susceptibility in PSC. METHODS: A total of 148 adult white northern European patients and 134 control subjects were studied. HLA DRB1, DQA1, DQB1 alleles and DRB1*04, DRB1*13 and DRB3 subtypes were determined by standard PCR-genotyping. RESULTS: The primary associations with the DRB3*0101--DRB1*0301--DQA1*0501--DQB1*0201 and DRB1*1301--DQA1*0103--DQB1*0603 haplotypes were confirmed (O.R. = 2.69, p < 0.0000025 and O.R. = 3.8, p < 0.0005). In addition the strong protective influence of the DRB1*04--DQB1*0302 haplotype was reaffirmed (O.R. = 0.26, p < 0.000025) and a previously unreported negative (i.e. protective) association with the DRB1*0701--DQB1*0303 haplotype was also demonstrated (O.R. = 0.15, p < 0.005). Further analysis suggested that susceptibility/resistance encoded by the second and third susceptibility haplotypes and by the two resistance haplotypes may be determined by specific amino acids at DQbeta-87 and DQbeta-55, respectively. PMID- 12765484 TI - The hepatitic/cholestatic "overlap" syndrome: an Italian experience. AB - BACKGROUND: Patients with hepatitic and cholestatic autoimmune liver disease ("overlap syndrome") represent a diagnostic and therapeutic challenge. AIM: To evaluate the prevalence of the "hepatitic/cholestatic overlap" in a large series of consecutive patients with cholestatic autoimmune liver disease. METHODS: We re evaluated the diagnosis of 235 patients with autoimmune liver disease, including 70 with type 1 autoimmune hepatitis (AIH), 142 with primary biliary cirrhosis (PBC), and 23 with primary sclerosing cholangitis (PSC), using the revised International Autoimmune Hepatitis Group (IAIHG) scoring system. Anti mitochondrial, anti-nuclear, anti-smooth muscle, anti-liver kidney microsomal type 1, anti-liver cytosol type 1, perinuclear anti-neutrophil nuclear and anti soluble liver antigen antibodies were evaluated in each patient. RESULTS: Ten patients (3 with a previous diagnosis of PBC and 7 of PSC) scored as "probable" or "definite" AIH. These patients did not have a specific autoantibody profile. CONCLUSIONS: Among patients with PBC, the occurrence of a PBC/AIH overlapping syndrome is rare (2.1%), whereas among patients with PSC an overlap between PSC and AIH is frequent (30.4%). Whether patients with the hepatitic/cholestatic overlap syndrome would benefit from a combination therapy with immunosuppression and ursodeoxycholic acid remains to be established. PMID- 12765485 TI - Inhaled corticosteroids in childhood asthma: long-term effects on growth and adrenocortical function. AB - Inhaled corticosteroids (ICS) are the most potent of all the available inhaled treatments, and are effective medications for long-term control of asthma. However, their use in children is limited by the risk of systemic adverse effects. Although results reported in the literature on the adverse effects of ICS are conflicting and often restricted to a small number of cases with a limited follow-up, most of them show an early decrease in growth velocity without significant influence on final adult height. Partial adrenal suppression has also been demonstrated in children treated with ICS for more than 2 months. Only children with mild persistent, moderate, or severe asthma not controlled by non corticosteroid drugs should be treated with ICS for long periods. The dose of ICS must be individually adjusted to minimize the possible adverse effects on growth, and all children with asthma receiving long-term treatment with ICS must be regularly evaluated for growth impairment, which may necessitate dose reduction or drug replacement. PMID- 12765486 TI - Diagnosis, treatment, and prevention of Lyme disease in children. AB - The approaches to diagnosing and treating Lyme disease (LD) have been improved and refined as a result of basic and clinical research, and considerable practical experience. In addition, there have been recent studies that have allowed improvements in the ability to prevent infection with Borrelia burgdorferi. This paper will review the relevant literature and address recent developments in the diagnosis, treatment, and prevention of LD. Issues specifically related to the management of children will be identified. Controversies regarding treatment approaches will be examined in some detail. Understanding the clinical manifestations, or stage, of LD is crucial when approaching both diagnosis and treatment. Early localized disease is best diagnosed by recognizing the characteristic skin lesion, erythema migrans. Early disease will frequently, but not always, be accompanied by a detectable antibody response, particularly IgM antibody to the spirochete. Late disease, chiefly arthritis, is generally associated with high levels of IgG antibody. Western blot technology allows confirmation of enzyme immunoassay results and is especially useful when the latter is in the low or equivocal range. Early localized disease responds well to oral antibacterial therapy. Early disseminated disease, often associated with neurologic findings, may require parenteral therapy. The arthritis associated with LD frequently responds to oral antibacterials, but some refractory cases may require intravenous therapy, and occasionally surgery. Doxycycline is the oral antibacterial of choice, while amoxicillin and cefuroxime axetil are alternatives that may be preferred in young children. Owing to its long half-life and once daily dose administration, intravenous ceftriaxone has become the accepted standard for parenteral therapy. Tick avoidance has long been the mainstay for preventing LD. Antibacterial prophylaxis, using doxycycline, for tick bites has been shown to be an effective approach to prevention, but its relevance to pediatrics is uncertain. Vaccines designed to prevent infection have also been developed. PMID- 12765487 TI - Postoperative shivering in children: a review on pharmacologic prevention and treatment. AB - Postoperative shivering consists of muscular tremor and rigidity. It is often associated with body heat loss, although hypothermia alone does not fully explain the occurrence of shivering. Shivering is self-limiting, never becomes chronic, and is rarely associated with major morbidity. However, it affects the comfort of the patients, and may sometimes lead to more serious complications. The efficacy of a great variety of pharmacologic interventions to prevent shivering and to treat established symptoms has been tested in randomized controlled trials. These can be gathered systematically; recommendations on prevention and treatment can then be based on the strongest evidence. Unfortunately all these trials have been performed in adults. Thus, recommendations for the control of postoperative shivering in children have to be extrapolated from adult data. In adults, a systematic review strongly suggests that simple measurements are efficacious for both prevention and treatment. For prevention, extrapolation of these adult data indicates that three children have to receive intravenous clonidine 1.5 micro g/kg during anesthesia for one not to shiver, when they would have done so had they not received clonidine. For this degree of efficacy, the expected incidence of shivering (baseline risk) has to be high (approximately 50%). For treatment, extrapolation from adult data indicates that less than two children need to receive intravenous meperidine (pethidine) 0.35 mg/kg, or clonidine 1.5 micro g/kg for one to stop shivering five minutes after drug administration, when they would not have done so had they not received one of these drugs. Since the treatment of established shivering is efficacious, simple, inexpensive, and relatively safe, and since prevention is only efficacious if the baseline risk is very high, we recommend the 'wait and see' strategy. PMID- 12765489 TI - Atomoxetine. AB - Atomoxetine, formerly tomoxetine, is a selective norepinephrine reuptake inhibitor and a new, nonstimulant treatment for attention deficit hyperactivity disorder (ADHD). In vitro, ex vivo and in vivo studies have shown that atomoxetine is a highly selective antagonist of the presynaptic norepinephrine transporter with little or no affinity for other noradrenergic receptors or other neurotransmitter transporters or receptors. In four randomized, placebo controlled clinical trials conducted over 6-9 weeks in children and adolescents with ADHD, atomoxetine (total daily dose 1-1.8 mg/kg administered in one or two doses daily) reduced symptoms (hyperactivity, impulsiveness and inattention) as determined by the reduction in ADHD total score (34-38% with atomoxetine versus 13-15.7% with placebo [p < 0.05]). Atomoxetine also significantly improved ADHD subscale rating scores (p < 0.05 and p < 0.001), psychosocial well-being (p < 0.05) and ADHD-related problem behavior according to parent and teacher ratings (p < 0.05). Atomoxetine was well tolerated in clinical trials and discontinuation rates due to adverse events were low (<5%). The most common treatment-related adverse event was decreased appetite. Atomoxetine shows no abuse potential and is not a controlled substance in the US. PMID- 12765497 TI - Debriefing: care and sympathy are not enough. PMID- 12765493 TI - Linezolid: in infants and children with severe Gram-positive infections. AB - Linezolid is an oxazolidinone antibacterial agent that has inhibitory activity against a broad range of Gram-positive bacteria, including methicillin-resistant Staphylococcus aureus, vancomycin-resistant enterococci, and penicillin-resistant Streptococcus pneumoniae. The systemic clearance and, therefore, the area under the plasma concentration-time curve and elimination half-life of linezolid change with increasing age in pediatric patients. With the exception of pre-term neonates aged <1 week, systemic clearance is more rapid in pediatric patients aged 0-11 years than in adolescents. The pharmacokinetic profile of linezolid is similar in adolescents and adults. Linezolid was as effective as vancomycin in the treatment of pediatric patients with Gram-positive infections (clinical cure rate 89.3% vs 84.5%), and as effective as cefadroxil in the treatment of children and adolescents with Gram-positive uncomplicated skin and skin structure infections (91.0% vs 90.0%) in the clinically evaluable population of two randomized, comparator-controlled trials. The clinical cure rate with linezolid was 92.4% in a noncomparative trial in hospitalized pediatric patients with community-acquired pneumonia. All patients with proven pneumococcal pneumonia were considered cured. Linezolid is generally well tolerated. The most common drug-related adverse events in the comparator-controlled trials were diarrhea, nausea, and headache; most events were mild to moderate in severity. PMID- 12765496 TI - Current issues in Crohn's disease. PMID- 12765498 TI - Epitaph for the EBM in action series. PMID- 12765488 TI - Nonsteroidal anti-inflammatory agents in neonates. AB - The use of NSAIDs has become routine for adults and children in the management of pain. NSAIDs (other than aspirin [acetylsalicylic acid]) are also enjoying greater popularity as antipyretics since the recognition of Reye's syndrome's putative association with aspirin. In neonates, NSAIDs have been used for many years in an attempt to pharmacologically close the ductus arteriosus. This review examines the various NSAIDs and their potential and real applications in the neonatal population. For completeness, acetaminophen (paracetamol), which has weak NSAID activity and is a widely used analgesic and antipyretic in this patient group, was also included. The prostaglandin system is important for healthy development, and conversely there are unique risks posed by pharmacologic interference with this system in the neonatal period. The prostanoid system in neonates has the capacity to modulate nociception, but comes at the expense of interfering with nearly every organ system. Physiologic effects of inhibition of prostaglandin synthesis applicable to neonates include disruption of the sleep cycle, increased risk of pulmonary hypertension, alterations in cerebral blood flow, decreased renal function, disrupted thermoregulation, and alterations in hemostasis balance, among others. Prostaglandins are also important for the normal development of the central nervous, cardiovascular, and renal systems, and there is evidence that the proper genesis of these systems may be adversely effected by NSAID exposure in utero and in the neonatal period. Gastrointestinal adverse effects have provided the impetus for the development and marketing of selective cyclo-oxygenase type 2 (COX-2) inhibitors. These agents' reputation for safety in adults may not be applicable to neonates. COX-2 is involved in the development of several organ systems, and its inhibition may induce a prothrombotic state. The advent of parenteral formulations of cyclo-oxygenase inhibitors, including COX-2-selective agents, increases the therapeutic flexibility of NSAIDs. However, objective data on the safety of these agents have not kept pace with their clinical availability. PMID- 12765499 TI - Initial experience with capsule endoscopy at a major referral hospital. AB - OBJECTIVES: To determine the utility of capsule endoscopy in patients referred for investigation of suspected disease of the small intestine. DESIGN AND SETTING: Single centre, prospective, cohort study from 4 July 2001 to 8 September 2002. PATIENTS: Sixty consecutive patients who underwent capsule endoscopy for investigation of suspected disease of the small intestine. MAIN OUTCOME MEASURES: Abnormal findings at capsule endoscopy. RESULTS: Examination of the entire small bowel was achieved in 46/60 patients (77%). Thirty-two of 47 patients (68%) referred with obscure gastrointestinal bleeding had positive findings. Seven of nine patients (78%) referred for investigation of suspected Crohn's disease had small bowel erosions or ulcers consistent with Crohn's disease. The capsule was not passed in one patient. CONCLUSION: Capsule endoscopy is a novel, minimally invasive and useful tool for the investigation of the small intestine. PMID- 12765500 TI - Stress debriefing after childbirth: a randomised controlled trial. AB - OBJECTIVE: To test whether critical incident stress debriefing after childbirth reduces the incidence of postnatal psychological disorders. DESIGN: Randomised single-blind controlled trial stratified for parity and delivery mode. SETTING: Two large maternity hospitals in Perth. PARTICIPANTS: 1745 women who delivered healthy term infants between April 1996 and December 1997 (875 allocated to intervention and 870 to control group). INTERVENTION: An individual, standardised debriefing session based on the principles of critical incident stress debriefing carried out within 72 hours of delivery. MAIN OUTCOME MEASURES: Diagnosis of stress disorders or depression in the 12 months postpartum, using structured psychological interview and criteria of the Diagnostic and statistical manual of mental disorders, 4th edition. RESULTS: Follow-up information was available for 1730 women (99.1%), 482 of whom underwent psychological interview. There were no significant differences between control and intervention groups in scores on Impact of Events or Edinburgh Postnatal Depression Scales at 2, 6 or 12 months postpartum, or in proportions of women who met diagnostic criteria for a stress disorder (intervention, 0.6% v control, 0.8%; P = 0.58) or major or minor depression (intervention, 17.8% v control, 18.2%; relative risk [95% CI], 0.99 [0.87-1.11]) during the postpartum year. Nor were there differences in median time to onset of depression (intervention, 6 [interquartile range, 4-9] weeks v control, 4 [3-8] weeks; P = 0.84), or duration of depression (intervention, 24 [12-46] weeks v control, 22 [10-52] weeks; P = 0.98). CONCLUSIONS: There is a high prevalence of depression in women during the first year after childbirth. A session of midwife-led, critical incident stress debriefing was not effective in preventing postnatal psychological disorders, but had no adverse effects. PMID- 12765501 TI - Hepatitis C transmission and HIV post-exposure prophylaxis after needle- and syringe-sharing in Australian prisons. AB - OBJECTIVES: To determine whether infection with human immunodeficiency virus (HIV), hepatitis B virus (HBV) or hepatitis C virus (HCV) occurred after two potential episodes of exposure through needle- and syringe-sharing in Australian prisons, and to examine use of post-exposure prophylaxis (PEP) against HIV infection in the prison setting. DESIGN: Cohort study of potential contacts of two prisoners infected with HIV, HBV and HCV followed up for up to 14 months. SETTING: Two Australian prisons between November 2000 (time of exposure) and December 2001. PARTICIPANTS: Two index patients (both infected with HIV and HCV; one also infectious for HBV) from two different prisons, and 104 inmates who shared needles and syringes. MAIN OUTCOME MEASURES: Seroconversions to HIV, HBV and HCV related to the high-risk exposure and uptake and completion of HIV PEP determined from medical records of inmates. RESULTS: There were four seroconversions to HCV within 14 months of the potential exposure (14% of those susceptible in the cohort), but no recorded HIV or HBV seroconversions. Forty-six inmates (82% of those eligible) were offered PEP, and 34 of these (74%) elected to receive it. Only eight (24% of the 34) completed the full PEP course. CONCLUSIONS: HCV transmission in the prison setting is related to high-risk needle- and syringe-sharing. Administering HIV PEP in the prison setting is complicated by challenging risk assessment and follow-up. PMID- 12765502 TI - Urinary incontinence in subacute care--a retrospective analysis of clinical outcomes and costs. AB - OBJECTIVE: To investigate the effect of incontinence on clinical outcomes and costs for patients in subacute care. DESIGN: Retrospective analysis of data collected over a 3-month period in 1996. SETTING: 54 medical facilities in Australia and New Zealand providing subacute care in an inpatient setting. PATIENTS: 6773 episodes of care provided to 6455 rehabilitation and geriatric evaluation and management patients. MAIN OUTCOME MEASURES: Urinary continence status, treatment outcomes, length of stay, discharge destination, and nursing and allied healthcare costs. RESULTS: Discharge destination differed between incontinent and continent patients (57% compared with 82%, respectively, discharged home, and 29% compared with 12%, respectively, discharged to a nursing home or to further care). There was a difference in cost between patients who were continent and those who were incontinent throughout their episode of care (rehabilitation: $185.60 [95% CI, $181-$190] per day for incontinent and $156.82 [95% CI, $153-$160] for continent patients; and geriatric evaluation and management: $164.62 [95% CI, $157-$172] for incontinent and $121.40 [95% CI, $114 $129] for continent patients). However, multilevel analyses showed that, after allowing for age and level of functional independence, the contribution of continence status to the cost of care depended on the functional independence of the patient (cognitive function for orthopaedic patients [P < 0.01] and motor function for stroke patients [P = 0.04]). CONCLUSION: The relationship between continence status and cost of care is complex. However, the cost differences found in our study need to be considered in payment systems, allocation of staff levels on wards and in development of casemix classifications. PMID- 12765503 TI - Preventing local transmission of SARS: lessons from Singapore. PMID- 12765505 TI - Pulmonary arterial hypertension: a new era in management. AB - Pulmonary arterial hypertension (PAH) is a heterogeneous condition with a wide range of causes. The diagnosis is often delayed or missed. PAH is covert in its early stages, when its detection and treatment should have the most impact. Access in Australia to effective PAH therapies has lagged behind that in other affluent countries. New agents for PAH, now becoming available, improve symptoms and reduce pulmonary resistance, with some demonstrating an ability to reverse remodelling of the right ventricle. Best management of PAH is comprehensive and multidisciplinary. Centres of excellence are needed in geographically strategic areas. Aggressive efforts must be made to diagnose PAH and to facilitate access to effective therapies. PMID- 12765504 TI - Short-stay units and observation medicine: a systematic review. AB - OBJECTIVES: To conduct a systematic review of how short-stay observation units (SOUs) affect the efficiency of healthcare delivery and the quality of services provided. DATA SOURCES: MEDLINE, CINAHL, Best Evidence and The Cochrane Library were searched for the period 1 January 1960 to 31 July 2000. STUDY SELECTION: Studies were eligible if published in English and rated at National Health and Medical Research Council evidence levels I, II-1, II-2, or II-3; 12 comparative studies published between 1985 and 1998 met these criteria. DATA EXTRACTION: Data pertaining to clinical outcomes, length of stay, re-presentation rates, emergency department efficiency and costs of care were extracted and evaluated independently. DATA SYNTHESIS: As there was considerable heterogeneity in the patient populations and outcomes, results were summarised rather than subjected to meta-analysis. CONCLUSION: SOUs have the potential to increase patient satisfaction, reduce length of stay, improve the efficiency of emergency departments and improve cost effectiveness. However, SOUs have commonly been implemented alongside new clinical protocols, and it is not possible to distinguish the relative benefits of each. As demand increases, providing effective and cost-efficient care will become increasingly important. SOUs may help organisations that are attempting to streamline patient care while maintaining their quality of service delivery. PMID- 12765506 TI - Antiplatelet drugs. AB - Antiplatelet drugs protect against myocardial infarction, stroke, cardiovascular death and other serious vascular events in patients with a history of previous vascular events or known risk factors for cardiovascular disease. Aspirin reduces the risk of serious vascular events in patients at high risk of such an event by about a quarter and is recommended as the first-line antiplatelet drug. Clopidogrel reduces the risk of serious vascular events among high-risk patients by about 10% compared with aspirin. It is as safe as aspirin, but much more expensive. It is an appropriate alternative to aspirin for long-term secondary prevention in patients who cannot tolerate aspirin, have experienced a recurrent vascular event while taking aspirin, or are at very high risk of a vascular event (>/= 20% per year). Addition of clopidogrel to aspirin reduces the risk of serious vascular events among patients with non-ST-segment elevation acute coronary syndromes by 20%, and patients undergoing percutaneous coronary intervention by 30%, compared with aspirin alone. Addition of a glycoprotein IIb/IIIa receptor antagonist to aspirin reduces the risk of vascular events among patients with non-ST-segment elevation acute coronary syndromes by 10% and among patients undergoing percutaneous coronary intervention by 30%, compared with aspirin alone; it appears to provide incremental benefit in patients also treated with clopidogrel. Addition of dipyridamole to aspirin seems to be more effective than aspirin alone for preventing recurrent stroke, but its overall effect in preventing serious vascular events in patients with ischaemic stroke and transient ischaemic attack has not been determined. PMID- 12765507 TI - Phenytoin for the prevention of motion sickness. PMID- 12765508 TI - Which drug is most effective in treating childhood tinea capitis caused by Microsporum species? PMID- 12765509 TI - Recruitment to randomised studies. PMID- 12765510 TI - Ethical and legal issues and the "new genetics". AB - Although uniquely personal, the information from gene analyses impacts on parents, siblings, children and even entire ethnic groups. Doctors need to carefully balance the right of a patient to privacy against the wider family and society interests, consistent with ethical standards and their legal obligations. Doctors also need to be in a position to advise their patients of potential risks that may result from obtaining predictive genetic information, such as discrimination by third parties. While these issues are not new, they occur with new sharpness in the "new genetics", where clinicians have to be familiar not only with clinical significance, but also the ethical and legal implications of genetic analyses and information. PMID- 12765511 TI - Circumcision for phimosis and other medical indications in Western Australian boys. PMID- 12765512 TI - Treating phimosis. PMID- 12765513 TI - Metformin use as an adjunct to insulin treatment. PMID- 12765514 TI - Medical rosters and the Trade Practices Act. PMID- 12765517 TI - Caspases as regulators of apoptosis and other cell functions. AB - This review covers current knowledge on the involvement in apoptosis of a new family of endopeptidases denoted as caspases. Their structure, specific substrates, and inhibitors are considered. The recent classification of cysteine proteases of the caspase family based on their structural and functional features is presented. The biological significance of caspases not related to their proapoptotic effect is discussed. PMID- 12765518 TI - Consensus sequence of transcription factor SF-1 binding site and putative binding site in the 5' flanking regions of genes encoding mouse steroidogenic enzymes 3betaHSDI and Cyp17. AB - Using 42 nucleotide sequences extracted from the Transcription Regulatory Regions Database (TRRD) containing SF-1 transcription factor binding site, we have determined the decanucleotide (GTCAAGGTCA) consensus sequence for SF-1 binding. In the frequency matrix of this sequence nucleotides between the 3rd and the 7th position had the highest frequency and guanine nucleotides at the 6th and the 7th positions were recognized in all nucleotide sequences. The latter suggests a crucial role of these guanines for the interaction of DNA with SF-1 protein. The determined consensus and frequency matrix were used for search of putative SF-1 binding sites in regulatory regions of two genes, encoding mouse Cyp17 (17alpha hydroxylase/17-20-lyase) and 3betaHSDI (3beta-hydroxysteroid dehydrogenase/4delta 5delta-isomerase I), the microsomal enzymes involved in steroidogenesis. 5; Flanking regions of genes encoding Cyp17 and 3betaHSDI were shown to contain six and five such binding sites, respectively. The presence of the putative SF-1 binding sites in the regulatory regions of mouse Cyp17 and 3betaHSDI suggests that gene SF-1 could represent one of the putative genes which (as we predicted earlier) determine coordinated inheritable variability of hormonal activity in mouse Leydig cells. PMID- 12765519 TI - Ion transport coupled to terminal oxidase functioning in the extremely alkaliphilic halotolerant bacterium Thioalkalivibrio. AB - Proton transport in the terminal part of the respiratory chain in the extremely alkaliphilic halotolerant bacterial strain Thioalkalivibrio versutus was studied under near-optimum growth conditions (pH 9.0-9.5). Under these conditions, bacterial cells generated electric potential with the negative charge being inside the cells. When only the terminal part of the respiratory chain functioned, it was found that: 1) unlike other bacteria known, this bacterium did not acidify the medium in the presence of K(+) and valinomycin; 2) in the presence of an uncoupler, CCCP, but in the absence of valinomycin, reversible alkalinization of the medium occurred as a result of proton influx into the cells. Cyanide prevented this alkalinization. The difference spectra indicate that cell membranes contained cytochromes c and (b+o), some of which reacted with CO. The respiratory activity of membranes in the terminal part of the respiratory chain was optimal at pH 9.5 and specifically depended on sodium ions (C(1/2) = 10 mM). The data suggest the presence of a Na(+)-pump in the terminal part of the respiratory chain of the studied strain which can pump Na(+) out of the cells. PMID- 12765520 TI - Cyclosporin A-sensitive decrease in the transmembrane potential across the inner membrane of liver mitochondria induced by low concentrations of fatty acids and Ca2+. AB - At low Ca2+ concentrations the pore of the inner mitochondrial membrane can open in substates with lower permeability (Hunter, D. R., and Haworth, R. A. (1979) Arch. Biochem. Biophys., 195, 468-477). Recently, we showed that Ca2+ loading of mitochondria augments the cyclosporin A-dependent decrease in transmembrane potential (DeltaPsi) across the inner mitochondrial membrane caused by 10 micro M myristic acid but does not affect the stimulation of respiration by this fatty acid. We have proposed that in our experiments the pore opened in a substate with lower permeability rather than in the "classic" state (Bodrova, M. E., et al. (2000) IUBMB Life, 50, 189-194). Here we show that under conditions lowering the probability of "classic pore" opening in Ca2+-loaded mitochondria myristic acid induces the cyclosporin A-sensitive DeltaPsi decrease and mitochondrial swelling more effectively than uncoupler SF6847 does, though their protonophoric activities are equal. In the absence of P(i) and presence of succinate and rotenone (with or without glutamate) cyclosporin A either reversed or only stopped DeltaPsi decrease induced by 5 micro M myristic acid and 5 micro M Ca2+. In the last case nigericin, when added after cyclosporin A, reversed the DeltaPsi decrease, and the following addition of EGTA produced only a weak (if any) DeltaPsi increase. In P(i)-containing medium (in the presence of glutamate and malate) cyclosporin A reversed the DeltaPsi decrease. These data show that the cyclosporin A-sensitive decrease in DeltaPsi by low concentrations of fatty acids and Ca2+ cannot be explained by specific uncoupling effect of fatty acid. We propose that: 1) low concentrations of Ca2+ and fatty acid induce the pore opening in a substate with a selective cation permeability, and the cyclosporin A sensitive DeltaPsi decrease results from a conversion of DeltaPsi to pH gradient due to the electrogenic cation transport in mitochondria; 2) the ADP/ATP antiporter is involved in this process; 3) higher efficiency of fatty acid compared to SF6847 in the Ca2+-dependent pore opening seems to be due to its interaction with the nucleotide-binding site of the ADP/ATP-antiporter and higher affinity of fatty acids to cations. PMID- 12765521 TI - Localization of a site for interaction with hepatic male-specific proteins in two rat estrogen sulfotransferase genes. AB - Using electromobility shift assay the interaction of fragments of two paralogous rat estrogen sulfotransferase (Ste) genes with proteins of nuclear extracts from male and female rat liver was studied. Male-specific DNA-protein complexes were revealed with labeled oligonucleotides corresponding to fragments +1150/+1449, +1358/+1449, +1397/+1449, and +1417/+1449 of intron 1 of the Ste1 gene. The removal of a 20 bp region corresponding to the sequence +1430/+1449, or even either 5;- or 3;-terminal 5 bp of this region abolished the selective interaction of the oligonucleotides with the male-specific protein(s). According to the results of the experiments on mutual competition of the oligonucleotides, the fragment of the Ste2 gene corresponding to the sequence +1397/+1449 of the Ste1 gene formed complexes with the same male-specific protein(s) as the fragment of the Ste1 gene did. The data suggest the mapped element to participate in gender differentiation of the expression of the Ste1 and Ste2 genes. PMID- 12765522 TI - Protein content of human saliva in various psycho-emotional states. AB - The protein composition of human saliva depends on psycho-emotional state of individuals. Depression was accompanied by decrease of proteins of molecular masses ranging from 20 to 200 kD, whereas emotionally positive intellectual activity caused the opposite effect. It is suggested that human saliva may be used as an experimental model for the development of diagnostics of various psycho-physiological states. PMID- 12765523 TI - Comparative kinetic study of D-glucose oxidation by ruthenium(III) compounds catalyzed by FAD-dependent glucose oxidase and PQQ-dependent glucose dehydrogenase. AB - The comparative kinetic study of two glucose oxidizing enzymes, FAD-dependent glucose oxidase and PQQ-dependent glucose dehydrogenase, is presented in the artificial electron transfer mediator system based on ruthenium(III) compounds. It is demonstrated that FAD-dependent glucose oxidase and PQQ-dependent glucose dehydrogenase follow Michaelis kinetics in the D-glucose/ruthenium(III) system. PQQ-dependent glucose dehydrogenase is more active than FAD-dependent glucose oxidase in the process of D-glucose oxidation by ruthenium(III) compounds, this being due to the different catalytic mechanisms of these enzymes. PMID- 12765524 TI - Role of integrin alphavbeta3 in substrate-dependent apoptosis of human intestinal carcinoma cells. AB - Incubation of human intestinal carcinoma Caco-2 cells in suspension (i.e., in the absence of substrate contacts) leads to massive cell death by apoptosis. Since this type of apoptosis has been referred to as anoikis, we designated these cells as anoikis-positive. However, a minor proportion of Caco-2 cells, designated as anoikis-negative, survived in suspension. Extended incubation of the cells in suspension resulted in the reduction of the number of viable cells. In comparison to the original Caco-2 cell population, the anoikis-negative cells demonstrated markedly decreased levels of expression of integrin alphavbeta3 on the cell surface and of transcription of the alphav subunit gene. Activation of the signaling function of alphavbeta3 in the original Caco-2 cells led to substantial stimulation of anoikis, while the inhibition of expression of this receptor resulted in better resistance of the cells to anoikis. The data provide the first evidence that alphavbeta3 integrin can generate apoptosis-stimulating signals. PMID- 12765525 TI - A comparative study of the effect of abscisic acid and cAMP on protein synthesis in wheat caryopses under drought conditions. AB - The effect of exogenous abscisic acid and cAMP on synthesis of soluble proteins in wheat caryopses in drought has been studied. Both compounds affected the formation of the polypeptides whose synthesis was stimulated by dehydration: they increased the incorporation of the label into polypeptides of 13, 15, and 26 kD and decreased the incorporation of the label into polypeptides of 14, 64, and 77 kD. Abscisic acid and cAMP increased the level of the incorporation of [(14)C]leucine into the low-molecular-weight polypeptides of 12, 17, and 19 kD whose synthesis was suppressed by drought. These data suggest that the cyclic adenylate signal system is probably involved in the effect of abscisic acid on protein synthesis in drought. PMID- 12765526 TI - Bacillus species LU4 is an effective producer of thermostable site-specific endonuclease BspLU4I, an isoschizomer of AvaI. AB - The site-specific endonuclease BspLU4I was discovered in the thermophilic Bacillus species LU4 strain and purified to functionally pure state by chromatography on blue agarose, hydroxyapatite HTP, and heparin-Sepharose columns. Analysis of cleavage patterns of different DNAs with known nucleotide sequences demonstrated that the enzyme recognizes the CPyCGPuG site on the DNA. Cleavage points in the sequence were determined with the elongated primer method. It was shown that the endonuclease is an isoschizomer of AvaI. The final yield of the enzyme is 2.25.10(6) units per g wet biomass. PMID- 12765527 TI - Change in pH-dependent membrane insertion characteristics of trichosanthin caused by deletion of its last seven C-terminal amino acid residues. AB - Trichosanthin (TCS) is a type I ribosome-inactivating protein (RIP) that can selectively kill some types of cells at low concentration (0.1-1 nM). The pH dependent membrane insertion ability of TCS makes it possible that the internalized toxin avoids degradation in lysosomes and further undergoes transportation into the cytosol by some still unidentified mechanism. Here, we show that deletion of C-terminal residues affects interactions of modified TCS (C7-TCS) with lipids and reduces its pH-dependent membrane insertion ability. Fluorescence measurements indicate that at low pH C7-TCS undergoes profound conformational changes that causes exposure of a hydrophobic region and leads to oligomerization of the C7-TCS molecules. The results suggest that the membrane insertion of TCS at low pH might be important for translocation of TCS into the cytosol, which is important for exertion of the RIP activity of TCS. Deletion of the last seven C-terminal residues of TCS would reduce both its RIP activity in vitro and cytotoxicity in vivo, with the degree of decrease being more significant for the cytotoxicity in vivo. PMID- 12765528 TI - New structures of the O-specific polysaccharides of Proteus. 3. Polysaccharides containing non-carbohydrate organic acids. AB - Four new Proteus O-specific polysaccharides were isolated by mild acid degradation from the lipopolysaccharides of P. penneri 28 (1), P. vulgaris O44 (2), P. mirabilis G1 (O3) (3), and P. myxofaciens (4), and their structures were elucidated using NMR spectroscopy and chemical methods. They were found to contain non-carbohydrate organic acids, including ether-linked lactic acid and amide-linked amino acids, and the following structures of the repeating units were established: [Figure: see text], where (S)-Lac and (R)-aLys stand for (S)-1 carboxyethyl (residue of lactic acid) and N(epsilon)-[(R)-1-carboxyethyl]-L lysine ("alaninolysine"), respectively. The data obtained in this work and earlier serve as the chemical basis for classification of the bacteria Proteus. PMID- 12765529 TI - Intracellular peptidoglycan hydrolases of the bacterium Xanthomonas campestris XL 1. AB - A system of intracellular peptidoglycan hydrolases of Xanthomonas campestris XL-1 comprises about 10 enzymes of different localization and substrate specificity. Seven enzymes (A(1)-A(7)) are localized in cytosol, one enzyme (A(8)) in periplasm, and two enzymes (A(9), A(10)) were found in the fraction of cell walls and membranes. While the culture is entering the logarithmic growth stage from the stationary stage, a change occurs in the activity of the cytosolic enzymes: A(1) significantly increases, and A(5) and A(6) decrease. The spectrum of cytosolic enzymes also depends on the growth medium composition. The enzyme A(7) present in cells secreting extracellular enzymes (medium 5/5) was not found in non-secreting cells (LB medium). Unlike extracellular enzymes, intracellular peptidoglycan hydrolases are primarily acidic proteins. The data indicate that the system of intracellular peptidoglycan hydrolases of X. campestris is under complex and strict regulation. PMID- 12765530 TI - Effect of ethylene producer ethrel and antioxidant ionol (BHT) on the proteolytic apparatus in coleoptiles of wheat seedlings during apoptosis. AB - It was established that total proteolytic activity in etiolated wheat seedlings changes in ontogenesis in cycles: peaks of proteolytic activity correspond to the 3rd, 5th, and 8th days of seedling growth, respectively. The maximum of proteolytic activity preceded the maximum of nuclease activity, which may be due to activation of nucleases by proteolytic enzymes. According to inhibitory analysis the cysteine and serine proteases play the main role in apoptosis in wheat coleoptiles. Growing of seedlings in the presence of ethrel stimulated apoptosis in the coleoptile, and it increased (almost 6-fold) the proteolytic activity in its cells. On the other hand, the antioxidant ionol (BHT) suppressed the induction of proteases, particularly at the second stage of coleoptile development, and it slowed down the increase in the nuclease activity after 6th day of the seedling life. It is suggested that phytohormones and antioxidants participate in regulation of apoptosis in the ageing coleoptile, directly or indirectly effecting the proteolytic apparatus in the coleoptile cells. PMID- 12765531 TI - Role of reproductive hormones in control of apoptosis of T-lymphocytes. AB - Effects of chorionic gonadotropin (CG), estradiol, progesterone, and their physiological combinations on apoptosis of human peripheral blood T-lymphocytes were studied. Neither the hormones separately nor their combinations affected the spontaneous apoptosis of T-cells. On stimulation with mitogens, a high dose of CG (100 IU/ml) significantly increased apoptosis of T-lymphocytes, but its combination with steroid hormones specific for trimester I of pregnancy decreased this parameter. Apoptosis of T-lymphocytes induced by neutrophils in mixed culture was also inhibited by the hormone combination corresponding to trimester I. In greater detail, this hormonal combination was shown to display differential effects on different T-cell subpopulations: it stimulated apoptosis of CD8(+) lymphocytes (which seemed to be provided by CG) and inhibited apoptosis of CD4(+) cells. Apoptosis of T-lymphocytes induced by anti-CD95 was suppressed by a high dose of progesterone (100 ng/ml) and also by its combination with CG and estradiol specific for trimester III of pregnancy. Thus, the reproductive hormones studied effectively regulated apoptosis of peripheral blood T lymphocytes. The effect of the hormones depended on the cell type and their activation and seemed to be an important mechanism of hormonal control of immune reactions in pregnancy. PMID- 12765533 TI - Phorbol ester stimulates expression of the human tryptophanyl-tRNA synthetase gene. AB - The effect of the phorbol ester phorbol 12-myristate 13-acetate (PMA) on expression of the human interferon (IFN)-inducible tryptophanyl-tRNA synthetase (WRS) gene was studied. PMA caused an increase in the basal and IFNgamma-induced WRS protein content in HeLa and HEK293 cultured cells. Besides, PMA upregulated WRS mRNA level in HeLa cells. Since PMA is known as a selective activator of protein kinase C (PKC) and is widely used to study the PKC-related pathways, these results show possible PKC involvement in regulation of the WRS gene expression. PKC inhibition by staurosporine (10 and 100 nM) had no effect on either basal or IFNgamma-induced expression of WRS in either cell line. Consequently, PKC is not an indispensable element in WRS induction by IFNgamma. Rather, PKC may activate WRS gene expression only by a distinct pathway. PMID- 12765532 TI - A new binary system for photosensitized labeling of DNA polymerases in nuclear extract. AB - A binary system of reagents was used for photosensitized labeling of proteins of bovine testis nuclear extract. A dUTP analog containing 4-azido-2,5-difluoro-3 chloropyridyl group (FAP-dUTP) was used for the first time as a component of the binary system, and a dUTP analog containing the pyrenyl group (Pyr-dUTP) was used as a photosensitizer. Photoaffinity labeling of proteins of nuclear extract was performed using the radioactively labeled DNA duplex with the photoreactive FAP group at the 3;-end of elongating DNA strand and analog of the deoxyribose phosphate residue (3-hydroxy-2-hydroxymethyltetrahydrofuran (F) 5;-phosphate) at the 5;-end of the nick. Such structure is formed by the action of nuclear extract enzymes from the initial DNA duplex containing a synthetic apurine/apyrimidine site and is a photoreactive analog of a long-patch base excision repair intermediate. UV-irradiation modified a limited number of proteins of the nuclear extract. As shown using specific antibodies, the new binary system of photoreagents increases the efficiency of DNA polymerase beta labeling. PMID- 12765534 TI - Inhibitory effects of some flavonoids on the activity of mushroom tyrosinase. AB - Mushroom tyrosinase (EC 1.14.18.1) is a copper containing oxidase that catalyzes both the hydroxylation of tyrosine into o-diphenols and the oxidation of o diphenols into o-quinones, and then forms brown or black pigments. In the present study, the effects of some flavonoids on the oxidation of L-3,4 dihydroxyphenylalanine (L-DOPA) have been studied. The results show that flavonoids can lead to reversible inhibition of the enzyme. A kinetic analysis showed that the flavonols are competitive inhibitors, whereas luteolin is an uncompetitive inhibitor. The rank order of inhibition was: quercetin > galangin > morin; fisetin > 3,7,4;-trihydroxyflavone; luteolin > apigenin > chrysin. PMID- 12765535 TI - Nucleosome positioning on yeast plasmids is determined only by the internal signal of DNA sequence occupied by the nucleosome. AB - The possible role of border factors in determining the nucleosome positioning on a DNA sequence was investigated. To this end a family of recombinant plasmids based on Gal10Cyc1 promoter and neomycin phosphotransferase gene NPTII were created. A DNA sequence adjoining the GalCyc promoter was varied in these plasmids. Three nearly equally represented nucleosome positions on the GalCyc promoter were found. In the basal plasmid an FRT sequence adjoins the GalCyc promoter at the right. It contains an internal signal of multiple positioning. Its replacement with different DNA sequences does not affect nucleosome positioning on the GalCyc promoter. The nucleosome positioning on the GalCyc promoter does not depend on nucleosome positioning (or its absence) on adjoining sequences. The same is true for nucleosome positioning on FRT sequence. It was found also that nucleosomes' positioning on the NPTII gene and their mutual disposition, namely the spacing between neighboring nucleosomes (linker length) are determined by the location of positioning signals only. Generally the nucleosome positioning in our experimental model is determined solely by internal DNA sequence occupied by nucleosome. On the other hand, the action of this internal positioning signal does not extend to neighboring DNA sequences. PMID- 12765537 TI - Aniseikonia tests: valid = "true" but "true" is not equal to p < or =.05 or p anything. The NAT is useful. PMID- 12765538 TI - There are better ways today to measure and manage aniseikonia than only a gross gross estimation based on the refraction. PMID- 12765539 TI - The aniseikonia answer is "The Aniseikonia Inspector" CD; strabismus surgery problems; ocular "PT"; monovision followup. PMID- 12765540 TI - Quantification of incidental needle and suture contamination during strabismus surgery. AB - PURPOSE: To quantify the level of bacterial contamination of needles and sutures immediately after use in strabismus surgery. METHODS: Strabismus surgery was performed on 56 eyes from 31 pediatric strabismus surgical cases. Preoperative site preparation included instillation of 5% povidone-iodine in the conjunctival fornices in all cases. A total of 124 needles and 127 sutures were cultured immediately after final scleral passage. RESULTS: Seventeen of the 31 cases (54.8%) produced at least one positive specimen. Nineteen per cent of the needles and 25.2% of the sutures were culture positive. The bulk of positive specimens (96.7% of needles, 91.3% of sutures) produced 3 or less colony forming units, corresponding to 7 or less total viable organisms per needle or sutures in accordance with the dilution scheme. Coagulase-negative staphylococci overwhelmingly predominated. CONCLUSIONS: Needles and sutures used in strabismus surgery can become contaminated during surgery despite preoperative povidone iodine preparation. The number of viable bacterial contaminants is usually below the level known to consistently produce experimental endophthalmitis. However, a few needles and sutures carried a high contaminant load, suggesting the possibility that contaminated needles and sutures could potentially cause postoperative intraocular infection. PMID- 12765541 TI - Evaluation of a new direct-comparison aniseikonia test. AB - BACKGROUND AND PURPOSE: Aniseikonia is a condition in which the two eyes perceive images of different size or shape, causing a variety of visual symptoms including asthenopia. Besides anisometropes (with a prevalence of 5-10% in the population above age 20 years), also pseudophakes and refractive surgery patients are at risk. For example, 40% of the pseudophakes seem to suffer from aniseikonia. Reliable measurement and management of aniseikonia is therefore important. The "Aniseikonia Inspector" is a new, commercially available, software product to measure and manage aniseikonia. The purpose of this study is to evaluate this aniseikonia test of the Aniseikonia Inspector. METHODS: Aniseikonia was induced in four subjects, with normal vision, by means of afocal size lenses. Using the Aniseikonia Inspector, the resulting aniseikonia was measured in vertical, horizontal and diagonal directions. RESULTS: The average ratio between the measured aniseikonia and the induced aniseikonia was 0.98, 0.89 and 0.93 respectively for the vertical, horizontal and diagonal directions. For two consecutive measurements of the same aniseikonic state, the difference in measurement value was 97%, 75% and 94% of the time within one resolution step size (0.5% horizontally and vertically, and 0.7% diagonally). CONCLUSION: Aniseikonia was measured accurately. Measurements in the vertical direction were more accurate than in the diagonal or horizontal directions, which is probably due to fixation disparities. The Aniseikonia Inspector is a very useful new tool in treating the growing number of aniseikonia patients. PMID- 12765542 TI - Spontaneous recovery from inferior rectus contracture (consecutive hypotropia) following local anesthetic injury. AB - We present yet another case of apparent inferior rectus paralysis following retrobulbar anesthesia for cataract surgery. He initially had a typical course but, unusually, after developing hypotropia and presumed contracture, went on to spontaneously recover. We are unaware of any other such case described in the literature. Methods to enhance this more desirable course are considered. PMID- 12765543 TI - Followup: a case of acute loss of binocular vision and stereoscopic depth perception. (The misery of acute monovision, having been binocular for 68 years). AB - This is a continuation of a personal report by a knowledgeable vision scientist, of the loss of binocular vision due to a major vitreous hemorrhage. Please see the initial report for further details. (Romano PE. A case of acute loss of binocular vision and stereoscopic depth perception. (The misery of acute monovision, having been binocular for 68 years) Binocul Vis Strabismus Q 2003; 18:51-55). This report covers months three and four following the hemorrhage. In the two months since the initial report, typical daily problems from this handicap are described. Definite evidence of progressive resolution was finally seen at the end of the third month. The speed of this resolution is about 1% per day! PMID- 12765544 TI - The requirement of cytosolic phospholipase A2 for the PMA activation of proton efflux through the N-terminal 230-amino-acid fragment of gp91phox. AB - The absolute requirement for the 85 kDa cytosolic phospholipase A(2) (cPLA(2)) in the PMA stimulation of proton efflux through the NADPH-oxidase-associated proton channel, has previously been demonstrated using a PLB-985 cell line deficient in cPLA(2) (PLB-D). The flux of protons in Chinese-Hamster ovary (CHO) cells that express the N-terminal 230-amino-acid (NT) fragment of gp91(phox) is activated by arachidonic acid (AA) added externally. To investigate the physiological role of cPLA(2), and the intracellular AA that it releases, in the activation of proton flux through the NT fragment of gp91(phox), this fragment was stably expressed in PLB-985 cells (PLB-985 NT) and in PLB-D cells (PLB-D NT). The expression of the NT fragment of gp91(phox) by itself in PLB-985 did not initiate differentiation and did not alter their ability to undergo differentiation after the addition of DMSO. Addition of PMA induced a proton efflux from undifferentiated PLB-985 NT cells expressing the NT fragment of gp91(phox), which was inhibited by zinc. In contrast, PMA failed to activate proton efflux in undifferentiated PLB-D NT cells, lacking the expression of cPLA(2); however, addition of AA restored the efflux of protons in these cells. These results establish an essential and specific physiological requirement of cPLA(2)-generated AA in the activation of proton flux through the NT fragment of gp91(phox). PMID- 12765545 TI - Hydride transfer during catalysis by dihydrofolate reductase from Thermotoga maritima. AB - DHFR (dihydrofolate reductase) catalyses the metabolically important reduction of 7,8-dihydrofolate by NADPH. DHFR from the hyperthermophilic bacterium Thermotoga maritima (TmDHFR), which shares similarity with DHFR from Escherichia coli, has previously been characterized structurally. Its tertiary structure is similar to that of DHFR from E. coli but it is the only DHFR characterized so far that relies on dimerization for stability. The midpoint of the thermal unfolding of TmDHFR was at approx. 83 degrees C, which was 30 degrees C higher than the melting temperature of DHFR from E. coli. The turnover and the hydride-transfer rates in the kinetic scheme of TmDHFR were derived from measurements of the steady-state and pre-steady-state kinetics using absorbance and stopped-flow fluorescence spectroscopy. The rate constant for hydride transfer was found to depend strongly on the temperature and the pH of the solution. Hydride transfer was slow (0.14 s(-1) at 25 degrees C) and at least partially rate limiting at low temperatures but increased dramatically with temperature. At 80 degrees C the hydride-transfer rate of TmDHFR was 20 times lower than that observed for the E. coli enzyme at its physiological temperature. Hydride transfer depended on ionization of a single group in the active site with a p K(a) of 6.0. While at 30 degrees C, turnover of substrate by TmDHFR was almost two orders of magnitude slower than by DHFR from E. coli; the steady-state rates of the two enzymes differed only 8-fold at their respective working temperatures. PMID- 12765546 TI - Evaluation of the symmetric model for myosin-linked regulation: effect of site directed mutations in the regulatory light chain on scallop myosin. AB - Regulatory myosins are controlled through mechanisms intrinsic to their structures and can alternate between activated and inhibited states. However, the structural difference between these two states is unclear. Scallop (Pecten maximus) striated adductor myosin is activated directly by calcium. It has been proposed that the two heads of scallop myosin are symmetrically arranged and interact through their regulatory light chains [Offer and Knight (1996) J. Mol. Biol. 256, 407-416], the interface being strengthened in the inhibited state. By contrast, vertebrate smooth-muscle myosin is activated by phosphorylation. Its structure in the inhibited state has been determined from two-dimensional crystalline arrays [Wendt, Taylor, Trybus and Taylor (2001) Proc. Natl. Acad. Sci. U.S.A. 98, 4361-4366] and is asymmetric, requiring no interaction between regulatory light chains. Using site-directed mutagenesis of the scallop regulatory light chain, we have tested the symmetric model for scallop adductor muscle myosin. Specifically, we have made myosin hybrid molecules from scallop (P. maximus) myosin, in which the normal regulatory light chains have been replaced by expressed light chains containing mutations in three residues proposed to participate in the interaction between regulatory light chains. The mutations were R126A (Arg126-->Ala), K130A and E131A; made singly, in pairs or all three together, these mutations were designed to eliminate hydrogen bonding or salt linkages between heads, which are key features of this model. Functional assays to address the competence of these hybrid myosins to bind calcium specifically, to exhibit a calcium-regulated myofibrillar Mg-ATPase and to display calcium-dependent actin sliding were performed. We conclude that the symmetrical model does not describe the inhibited state of scallop regulatory myosin and that an asymmetric structure is a plausible alternative. PMID- 12765547 TI - Monitoring disappearance of monomers and generation of resistance to proteolysis during the formation of the activation domain of human procarboxypeptidase A2 (ADA2h) amyloid fibrils by matrix-assisted laser-desorption ionization-time-of flight-MS. AB - The term 'amyloidosis' is used to represent a group of protein misfolding diseases characterized by the polymerization of normally innocuous and soluble proteins or peptides into insoluble proteinaceous deposits. One of the several questions that remain unclear regarding the process of amyloid fibril formation is related to the status of the protein when such a process begins. Protein engineering is one of the selected approaches to study amyloidosis. Characterization of many variants of a protein can give information about why a soluble protein aggregates to form fibrils. In the present study, we report information on the conformational changes that precede the formation of fibrils, monitored by the complementary use of exoproteolysis and matrix-assisted laser desorption ionization-time-of-flight-MS. This is a novel application of an easy and fast approach. In addition, we used it to evaluate the ability of the model protein ADA2h (activation domain of human procarboxypeptidase A2) and their mutants to generate amyloid fibrils. It could be a useful test to screen protein variants and to study to what extent some physicochemical parameters affect fibrillogenesis. PMID- 12765566 TI - [Blind liver biopsy in current medicine]. PMID- 12765548 TI - Phospholipase activity on N-acyl phosphatidylethanolamines is critically dependent on the N-acyl chain length. AB - We have recently shown that an endogenous phospholipase A2 from bovine erythrocytes does not hydrolyse NAPEs (N-acyl L-alpha-phosphatidylethanolamines), which accumulate remarkably in this system [Florin-Christensen, Suarez, Florin Christensen, Wainszelbaum, Brown, McElwain and Palmer (2001) Proc. Natl. Acad. Sci. U.S.A. 98, 7736-7741]. Here we investigate the causes underlying this resistance. N-acylation of PE (L-alpha-phosphatidylethanolamine) results in alteration of charge, head-group volume and conformation, the last two features depending on the N-acyl chain length. To evaluate each effect separately, we synthesized NAPEs with selected N-acyl chain length. We found that phospholipase A2 has considerable activity against N-acetyl PE, but is poorly active against N butanoyl PE and only marginally active against N-hexanoyl PE, whereas the activity is completely lost when N-hexadecanoyl PE is presented as a substrate. On the other hand, N-hexanoyl PE does not inhibit phospholipase A2 activity, suggesting that this substrate fails to enter the hydrophobic channel. Phospholipase C presents a similar, but less sharp pattern. Molecular dynamics simulations of the polar head group of selected NAPEs reveal a substantially increased conformational variability as the N-acyl chain grows. This larger conformational space represents an increased impairment limiting the access of these molecules to the active site. Our data indicate that, whereas a change in charge contributes to diminished activity, the most relevant effects come from steric hindrance related to the growth of the N-acyl chain. PMID- 12765567 TI - [Oral diet in health personnel, fulfillment of daily recommendations]. AB - BACKGROUND: Nutritional education is an essential issue for the proper approach to patients. One of the first issues to have in mind is the appropriate nutritional guidelines in theses professionals in their routine life. The aim of out work was to determine the oral intake pattern in a group of health professionals. MATERIALS AND METHODS: A 24-hours nutritional survey (encuesta) was performed in a group of 51 health professionals (including both medical and nurse personnel) who attended a course in specialized training in nutrition at Hospital Universitario Rio Hortega. All participants had their weight, length, age, and profession recorded. Before complying with diet they received an identification session of alimentary rations. RESULTS: The mean age of participants was 25.9 (5.4) years and they were all women (52.9% nurses and 47.1% physicians). Body mass index (BMI) was 21.9 (2.6). The intake of vitamin D was lower than international recommendations (RDA): 2.25 (3.27) g/day. However, the intake of vitamin C, 183.8 (118) mg/day; vitamin B12, 4.73 (3.18) mg/day; folic acid, 262.4 (126) g/day, and iodine, 310.3 (185) g/day were higher than international recommendations. The caloric intake was appropriate for the age of participants and protein intake was 2-fold above normality 1.66 (0.9) g/ kg/day. Differences were observed only between the group of nurses and physicians regarding iodine intake, slightly higher in the former individuals. CONCLUSION: In summary, the caloric intake in these health professionals is appropriate, and there is an excess in the intake of proteins, vitamin C, vitamin B12, folic acid and iodine, with a low intake of vitamin D. PMID- 12765568 TI - [Application of a method for the early detection of diabetic retinopathy in Primary Health Care]. AB - PATIENTS AND METHODS: An ophthalmic examination was performed in 1,495 diabetic patients by means of a photographic method with a non-mydriatic retinal (Ffo-CNM) camera (with inclusion of 81.5% of the total diabetic population in our area attended in three primary care centers in Terrasa, Barcelona. RESULTS: The prevalence of diabetic retinopathy known prior to the study was 15% (n = 218). With the application of the Ffo-CNM method the prevalence increased up to 28% (p = 0.0001). The diagnoses obtained were: normal fundoscopic appearance (bilateral), 61% (n = 908), DR without maculopathy, 22% (n = 334); DR with maculopathy, 5% (n = 75); severe DR or preproliferative DR, 0.1% (n = 1), and proliferative DR, 0.7% (n = 10). The fundus was not visible in 11% (n = 167) of patients, mainly because of darkness refractory miosis (48%) and media opacity (43%). Thirty-nine percent of examined patients were referred to the referral Ophthalmology department either because DR or non-visible eye fundus. CONCLUSIONS: The use of Ffo-CNM enhances the ophthalmoscopic examination in the diabetic population given the simplicity of the technique and its accessibility with the corresponding increase in the number of diagnoses of DR. Likewise, it improves the care quality to the diabetic patient and also the reationalization of patient referral to the ophthalmologic centers. PMID- 12765569 TI - [Systemic embolism after reversion to sinusal rhythm of persistent atrial flutter]. AB - BACKGROUND: The incidence of embolism in atrial flutter has been underestimated in the routine clinical practice. PATIENTS AND METHODS: In this study the incidence of thromboembolic events after restoration of sinus rhythm (by catheter ablation or cardioversion) was compared in two groups of consecutive patients, with a different anticoagulation protocol. A total of 169 patients were evaluated. A first retrospective analysis of 79 non anticoagulated patients (group I). A second prospective group of 90 patients who were treated with an anticoagulation protocol (group II) similar to that for patients with atrial fibrillation. All had typical atrial flutter of at least one month's duration before the procedure. RESULTS: The mean age of patients in group I was 61 12 years and the mean left ventricular ejection fraction was 57 6%. Patients in group II had a mean age of 61 10 years and the mean left ventricular ejection fraction was 56 9%. No differences were observed regarding prevalence of structural cardiopathy, arterial hypertension, diabetes mellitus, left ventricular dysfunction, atrial size or atrial fibrillation between the two groups of patients. Four patients in the retrospective analysis (5%) had an embolic event associated with the procedure, compared with 0 (0%) in the group of patients treated with the anticoagulation protocol. The efficient anticoagulation was associated with a lower risk of thromboembolic events (p < 0.05). CONCLUSIONS: The incidence of embolic events after reversion to sinusal rhythm of persistent atrial flutter can be decreased. These patients should follow the same recommendations of anticoagulation that apply for patients with persistent atrial fibrillation that are going to be reverted to sinus rhythm. PMID- 12765570 TI - [Complications and yield of percutaneous liver biopsy with ultrasonographic marking]. AB - OBJECTIVES: To describe the complications and the diagnostic yield of percutaneous liver biopsies performed under ultrasonographic control in an Internal Medicine department. PATIENTS AND METHODS: A total of 750 percutaneous liver biopsies were performed at the Internal Medicine department of our institution during the 1995-2001 time period. The Menghini technique was employed previous ultrasonographic marking of the puncture site was performed by the same physicians who performed the biopsy. Major and minor complications as well as the diagnostic yield (obtention of diagnostic hepatic tissue) of the technique were recorded. RESULTS: No deaths occurred in our series. Two patients (0.3%) had major complications presenting as liver subcapsular hematoma with hemoperitoneum which resolved with conservative measures. One of them was a male patient with acute leukemia and the other was a woman on antiaggregant therapy. Eight patients (1.1%) had minor complications (vasovagal syncope), all of them young males. In three cases no adequate material for diagnosis was obtained (technique yield 99.6%). CONCLUSIONS: In our series, percutaneous liver biopsy with ultrasonographic marking is a diagnostic technique associated with a low rate of severe complications and with a high diagnostic yield. The use of ultrasonographic marking by the same physicians performed the biopsy does not involve a longer time or increased cost of the technique, provides more safety in the procedure and has a low morbidity rate. This procedure could be used on a routine basis for all percutaneous liver biopsies. PMID- 12765571 TI - [Treatment of coagulopathy in severe sepsis]. PMID- 12765572 TI - [Current Internal Medicine. An interested vision]. PMID- 12765573 TI - [Treatment of penicillin-resistant pneumococcal infections in adults]. PMID- 12765574 TI - [Treatment of systemic vasculitis]. PMID- 12765575 TI - [Bilateral mechanical gonalgia in a middle-aged woman]. PMID- 12765576 TI - [Fifty-four year old woman with acute neuralgia in the D4 dermatome]. PMID- 12765577 TI - [Relapsing cutaneous lesions and cervical lymph node enlargment in a 72-year old woman]. PMID- 12765578 TI - [Manifestations of liver disease caused by hepatitis C virus]. PMID- 12765580 TI - [Anosmia as presenting symptom of Churg-Strauss syndrome]. PMID- 12765581 TI - [Reversible posterior leukoencephalopathy syndrome in an HIV-positive patient]. PMID- 12765582 TI - [SemFYC Summer School]. PMID- 12765583 TI - [An approach to home care. How does the Catalonian public health system handle home care?]. AB - OBJECTIVE: To describe the organizational model, resources and implementation of the home care program run by the Catalonian public health service, and to note difficulties reported by primary care professionals. DESIGN: Cross-sectional, descriptive study.Setting. Autonomous Community of Catalonia. PARTICIPANTS: 214 primary care centers (PCCs) operating under reformed administrative procedures were surveyed, and responses were obtained from 112 (52.3%). MAIN MEASURES: Semistructured, self-administered questionnaire sent by mail, with variables that identified the center, resources, organization and use of protocols for home care. Questionnaires were collected during July to November, 2000. RESULTS: Home care programs were being used at 90.2% (95% CI, 84.7%-95.7%) of the centers, and had been in effect for a mean of 5.6 years. Nursing services predominated in 64% of the case, with nurses spending a mean of 5.09 h/week on the program, twice as much time as physicians and social workers. The mean rate of computerization of the data was 31.3%, and was highest (P<.05) in Gerona (51%) and in PCCs run privately (70%). Of all participating PCCs, 70% had social workers on the staff, 13% had a home care nurse and 50% operated in coordination with social services. Of all PCCs, 79.5% (95% CI, 72.1%-86.9%) used specific protocols for pressure sores (69.1%), terminal illnesses (43.6%), bedridden patients (41.8%) and pain management (40.9%). Evaluations were done with the Barthel scale (73.2%), the Mini-Mental State Examination (73.2%) and the Norton scale (53.6%). Continuing education (66%) and activities for care providers (>75%) were frequent in home care programs. The main difficulties identified were the burden of care (65.2%), time constraints (51.8%), inadequate social support (43.8%) and lack of coordination with other levels of care (33%). CONCLUSIONS: Noteworthy findings were the dedication of nursing staff to the home care programs, the low level of computerization of the data, the limitations in social service resources, the uniformity of assessment protocols and scales, and the agreement regarding current difficulties. PMID- 12765584 TI - [Commentary: reflections on the reality of home care]. PMID- 12765585 TI - [Primary care resources use variation by under seven year old children in a rural and an urban area. Cohort study]. AB - OBJECTIVE: [corrected] To describe differences between two health districts in primary care resource use. DESIGN: Retrospective population cohort study. SETTING: A rural healthdistrict (RD) and an urban health district (UD). PARTICIPANTS: A cohort of children born in 1991 in a UD (n=104) and a cohort of children born in 1991 and 1992 in an RD (n=88), registered in their primary care centres between 0 and 6 years old. MAIN MEASUREMENTS: Attention episodes, requested visits, practicioner programmed visits, nursery programmed visits and diagnosis tests. RESULTS: Significative differences were noticed in the attention episode average(ZU=34.0; 95% CI, 31.5%-36.5%); (ZR=25.6; 95% CI, 22.7%-28.5%), in requested visits (ZU=45.8; 95% CI, 41.8%-49.9%); (ZR=28.8; 95% CI, 25.3%-31.9%), in practicioner programmed visits (ZU=7.1; 95% CI, 6.6%-7.5%), (ZR=11.2; 95% CI, 10.7%-11.7%) and in nursery programmed visits (ZU=9.8; 95% CI, 9.0%-10.6%) (ZR=12.0; 95% CI, 11.4%-12.5%). At least one diagnosis test was applied to 90.3% of children in the UD and to 64.8% of children in the RD; and in the UD for each attention episode 1.32 (95% CI, 1.28%-1.36%) requested visits were attended to and 0.2 (95% CI, 0.17%-0.23%) diagnosis tests were made; whereas in the RD 1.1 (95% CI, 1.08%-1.13%) requested visits and 0.09 (95% CI, 0.06%-0.12%) diagnosis tests. In both health districts the average of attention episodes, requested visits and tests was significantly higher in children who had been hospitalized. CONCLUSIONS: Primary care services utilization and the number of diagnosis test and attention episode visits are higher in the urban district than in the rural one. PMID- 12765586 TI - [Factors conditioning pulse pressure in type-2 diabetics in a primary care population suffering from hypertension]. AB - OBJECTIVE: To analyse the differences in pulse pressure (PP) between type-2 diabetic and non-diabetic hypertense patients, using clinical blood pressure (CBP) measurement and out-patient blood pressure monitoring (OPBPM) over 24 hours in primary care. DESIGN: Cross-sectional descriptive study. SETTING: Primary care. PARTICIPANTS: 163 hypertense patients in a random sample, including the first patient of each day with an appointment. MAIN MEASUREMENTS: CBP (Hg sphygmomanometer) and OPBPM recording. PP was considered high when the difference between systolic and diastolic pressure was over 60 mm Hg. RESULTS: 31.3% (51 patients) were diabetic. Mean clinical PP was 75.9 +/- 18.4 in diabetics and 64.5 +/- 18.9 mm Hg in non-diabetics (P<.001). Mean out-patient PP was 61.4 +/- 13.8 in diabetics and 53.5 +/- 11 mm Hg in non-diabetics (P<.001), with clinical and out-patient PP correlating with age. Clinical and out-patient PP were high in 63.8% and 57.4%, respectively, of hypertense diabetics. The logistical regression model showed more likelihood of high PP in those over 64 and in those with isolated systolic hypertension and/or diabetes mellitus. CONCLUSIONS: We found considerable differences between clinical and out-patient PP, and between the PP of the diabetic and non-diabetic population with hypertension. High out-patient PP in hypertense patients is related to diabetes and isolated systolic hypertension. PMID- 12765587 TI - [Effect of type-2 diabetes mellitus on the quality of life of patients treated at primary care consultations in Spain]. AB - OBJECTIVES: To evaluate the impact of type-2 diabetes mellitus (DM2) on the health-related quality of life (HRQL) of patients and to analyse the impact of social-demographic and clinical variables and of resource use. DESIGN: Cross sectional study with retrospective and prospective information gathering (HRQL questionnaire). Setting. 29 primary care centres from the whole of Spain. PARTICIPANTS: 1041 patients diagnosed with DM2 after the age of 30, chosen at random from patients' records at participating centres. Main measurements. HRQL evaluated through the general questionnaire EQ-5D, which enabled HRQL of patients to be compared with that of the general Spanish population, obtained from a sample of 8963 people. RESULTS: Patients with DM2 had worse HRQL (mean EVA index of 0.71) than people in the general population of the same age and gender (mean EVA index of 0.81). Women, older patients and obese patients had worse HRQL. In clinical terms, patients with some DM2-related complication, deficient glycaemic control and in receipt of insulin treatment had worse HRQL than patients without complications, acceptably controlled patients or those receiving non pharmacological or oral anti-diabetes treatment. CONCLUSIONS: DM2 is associated with worse HRQL for patients, and more so for patients with complications, poor control of glucaemia or under insulin treatment. PMID- 12765588 TI - [Prescription of non-steroidal anti-inflammatories and gastric protectors in emergency service]. AB - OBJECTIVES: To find the number of patients seen in casualty who received gastropathy prophylaxis with NSAIDs that complied with the pharmaco-therapeutic guidelines of the health district. DESIGN: Descriptive study of technical scientific quality. SETTING: The Casualty Department of the Ramon y Cajal hospital. PARTICIPANTS: Randomised sample of 400 emergencies attended in January 1999 and 2000, excluding paediatric cases, people previously treated with gastro protectors, patients admitted and the deceased. Main measurements. On the basis of casualty records: suitability of indication and criteria used; if unsuitable, how; adequacy of selection of the principle, patient' age and sex, digestive pathology history, area and year of care. RESULTS: NSAIDs were prescribed in 37.9% of cases (33.27-42.79), with prescription unsuitable in 21.19% (15.2-28.3). 15% (9.5-22) were unsuitable by default; and 54.2% (35.1-72.1), by excess. Unsuitability was greater among women, if there was history of digestive pathology, and among older people, with no differences found for area or year of care. The choice of gastric protector was not suitable in any case. CONCLUSIONS: Given the amount of primary care prescription originating in hospital casualty departments, and its effect on the training of family medicine interns, strategies coordinated between health care levels must be introduced in order to improve gastric protection prescription in NSAID gastropathy prophylaxis. The effectiveness of these strategies must be evaluated. PMID- 12765589 TI - [Influence of social support on patients with essential hypertension]. AB - OBJECTIVE: To determine whether social support affects blood pressure control in patients diagnosed with essential Hypertension. DESIGN: Observational, prospective study, with a year's follow-up. SERRING: Marinamansa-A Cuna Health Centre, Ourense (Galicia), Spain. PARTICIPANTS: 236 patients diagnosed with essential hypertension in health centre controls. MEASUREMENTS: During the one year follow-up the following clinical variables were measured: age, sex, blood pressure, severity of hypertension, tobacco consumption, alcohol consumption, presence of diabetes mellitus, hypercholesterolaemia, Body Mass Index and compliance with treatment. Social and family variables were: marital status, cultural level, economic and social status, type of family, stressful vital events and social support. RESULTS: Mean age was 63.51 (62.05-64.96); 66.1% were women. Predominant kind of family was nuclear (64.3%). 30.2% had low social network (0-1 social contacts). We found 22% poor functional social support. Social support remained stable throughout the study. Hypertense patients with poor social networks had, after control for possible confusing variables, their systolic pressure 9.59 mm Hg (2.6716,51) and diastolic pressure 4.29 mm Hg (0.448.15) higher than hypertense patients with broader social networks. CONCLUSION: Hypertense patients with a poor social network had higher blood pressure figures than hypertense patients with wider social networks. PMID- 12765590 TI - [Clinical management units in primary care: a question under debate]. AB - OBJECTIVE: To find the views of health professionals on how the terms of the new management agreement with clinical units affect the quality of health delivery. DESIGN: Qualitative study to find professionals' views. Individual opinions of key informers were obtained, secondary sources were analysed and a discussion group rounded off the study. The categories for the analysis were established on the basis of the results obtained. SETTING: Primary care districts in the provinces of Sevilla and Cadiz, in the Autonomous Community of Andalusia. Health districts of differing social-economic and geographical characteristics were chosen. PARTICIPANTS: 9 professionals involved in the new system of management. The selection criteria refer to the kind of professional, the unit or work team and the characteristics of the health district. METHOD: The semi-structured interview with key informers and a discussion group were the basic techniques for gathering information. RESULTS: A view that the clinical management units were designed as a savings strategy was detected. There were differences about primary care teams. CONCLUSIONS: Professionals think there is too much rigidity and adding-up in the measurement of targets and objectives. This creates unfavourable working conditions, which affects the quality of health service delivery. PMID- 12765591 TI - [Information systems in primary care: must we codify with the CIE-9-MC?]. PMID- 12765592 TI - [Stress at work: a new risk factor. What do we know and what can we do?]. PMID- 12765593 TI - [Surveys as a research technique. Composition of questionnaires and statistical processing of data (I)]. PMID- 12765594 TI - [Triple-blind clinical trial with placebo control to evaluate the efficacy of a heparin of low molecular weight (bemiparin) for treating slow-responding ulcers in diabetic foot in primary care]. AB - OBJECTIVES: To establish the degree of efficacy of bemiparin treatment over 3 months in the improvement of slow-responding ulcers in diabetic foot. Also, to evaluate the safety of bemiparin and quality of life and to compare the evolution of retinopathy and nephropathy against placebo. DESIGN: Stage III clinical trial to evaluate efficacy and safety in a new indication of a medicine already on the market, parallel in two branches, randomised, triple-blind, and controlled with placebo. SETTING: Health care centres in Mallorca, Spain. PARTICIPANTS: 42 patients per branch, over 18, with type-1 or 2 DM of over 3 years evolution, and one or more first or second-degree ulcers on the Wagner scale, distal to the knee, that did not heal in three months of health care. Randomised allocation in blocks of four.Interventions. The experimental drug was bemiparin (heparin of low molecular weight), injected subcutaneously at 3500 IU/day for the first 10 days and 2500 IU/day up to 90 days. As control, physiological serum was injected sub cutaneously in a similar volume for masking. MAIN MEASUREMENTS: An "effect"was defined as a reduction of at least 50% in its surface area and/or a favourable evolution in status to a degree between the control at the start of treatment and at three months. Other measurements included proteinuria, retinography and quality of life (SF-36). Analysis of efficacy through principle of intention to treat. PMID- 12765595 TI - [Self-measurement of blood pressure in primary care (I)]. PMID- 12765596 TI - [Urinary infection by Aerococcus urinae]. PMID- 12765597 TI - [Restless legs syndrome. Is it recognised in primary care?]. PMID- 12765598 TI - [An infrequent case of nasal obstruction]. PMID- 12765599 TI - [Local cutaneous relapse in breast cancer]. PMID- 12765600 TI - [Not confusing estimates with reality]. PMID- 12765601 TI - [A research project that was unnecessarily exclusive in practice: restriction on participation depending on computer back-up]. PMID- 12765602 TI - Hints of a therapeutic vaccine for Alzheimer's? PMID- 12765603 TI - Learning is bitter and sweet in ventral striatum. PMID- 12765604 TI - Synaptotagmin in Ca2+ -dependent exocytosis: dynamic action in a flash. AB - Synaptotagmins have been the popular candidates for the Ca2+ sensor that couples local rise in Ca2+ to neurotransmitter release. Studies in worm, fly, and mouse corroborate the likely role for synaptotagmin I, the best-studied synaptotagmin prototype, as a Ca2+ trigger for synaptic vesicle exocytosis. Recent investigations have focused on structural domains of synaptotagmin that are critical for its function. Here we provide a brief overview of synaptotagmin I and discuss recent studies within the framework of neurotransmitter release mechanisms for fast synaptic transmission. PMID- 12765605 TI - Beauty in the brain of the beholder. PMID- 12765606 TI - New paradigm for optical imaging: temporally encoded maps of intrinsic signal. AB - We present a new technique for acquiring and analyzing intrinsic signal optical images of brain activity, using continuous stimulus presentation and data acquisition. The main idea is to present a temporally periodic stimulus and to analyze the component of the response at the stimulus frequency. Advantages of the new technique include the removal of heart, respiration, and vasomotor artifacts, a dramatic increase in spatial resolution, and a 30-fold or greater reduction in acquisition time. We also present a novel approach to localizing instantaneous neuronal responses using time-reversed stimuli that is widely applicable to brain imaging. To demonstrate the power of the technique, we present high-resolution retinotopic maps of five visual areas in mouse cortex and orientation maps in cat visual cortex. PMID- 12765607 TI - Antibodies against beta-amyloid slow cognitive decline in Alzheimer's disease. AB - To test whether antibodies against beta-amyloid are effective in slowing progression of Alzheimer's disease, we assessed cognitive functions in 30 patients who received a prime and a booster immunization of aggregated Abeta(42) over a 1 year period in a placebo-controlled, randomized trial. Twenty patients generated antibodies against beta-amyloid, as determined by tissue amyloid plaque immunoreactivity assay. Patients who generated such antibodies showed significantly slower rates of decline of cognitive functions and activities of daily living, as indicated by the Mini Mental State Examination, the Disability Assessment for Dementia, and the Visual Paired Associates Test of delayed recall from the Wechsler Memory Scale, as compared to patients without such antibodies. These beneficial clinical effects were also present in two of three patients who had experienced transient episodes of immunization-related aseptic meningoencephalitis. Our results establish that antibodies against beta-amyloid plaques can slow cognitive decline in patients with Alzheimer's disease. PMID- 12765608 TI - Cdk5 is a key factor in tau aggregation and tangle formation in vivo. AB - Tau aggregation is a common feature of neurodegenerative diseases such as Alzheimer's disease, and hyperphosphorylation of tau has been implicated as a fundamental pathogenic mechanism in this process. To examine the impact of cdk5 in tau aggregation and tangle formation, we crossed transgenic mice overexpressing the cdk5 activator p25, with transgenic mice overexpressing mutant (P301L) human tau. Tau was hyperphosphorylated at several sites in the double transgenics, and there was a highly significant accumulation of aggregated tau in brainstem and cortex. This was accompanied by increased numbers of silver-stained neurofibrillary tangles (NFTs). Insoluble tau was also associated with active GSK. Thus, cdk5 can initiate a major impact on tau pathology progression that probably involves several kinases. Kinase inhibitors may thus be beneficial therapeutically. PMID- 12765609 TI - Unwrapping glial biology: Gcm target genes regulating glial development, diversification, and function. AB - Glia are the most abundant cell type in the mammalian brain. They regulate neuronal development and function, CNS immune surveillance, and stem cell biology, yet we know surprisingly little about glia in any organism. Here we identify over 40 new Drosophila glial genes. We use glial cells missing (gcm) mutants and misexpression to verify they are Gcm regulated in vivo. Many genes show unique spatiotemporal responsiveness to Gcm in the CNS, and thus glial subtype diversification requires spatially or temporally restricted Gcm cofactors. These genes provide insights into glial biology: we show unc-5 (a repulsive netrin receptor) orients glial migrations and the draper gene mediates glial engulfment of apoptotic neurons and larval locomotion. Many identified Drosophila glial genes have homologs expressed in mammalian glia, revealing conserved molecular features of glial cells. 80% of these Drosophila glial genes have mammalian homologs; these are now excellent candidates for regulating human glial development, function, or disease. PMID- 12765610 TI - Topographic motor projections in the limb imposed by LIM homeodomain protein regulation of ephrin-A:EphA interactions. AB - The formation of topographic neural maps relies on the coordinate assignment of neuronal cell body position and axonal trajectory. The projection of motor neurons of the lateral motor column (LMC) along the dorsoventral axis of the limb mesenchyme constitutes a simple topographic map that is organized in a binary manner. We show that LIM homeodomain proteins establish motor neuron topography by coordinating the mediolateral settling position of motor neurons within the LMC with the dorsoventral selection of axon pathways in the limb. These topographic projections are established, in part, through LIM homeodomain protein control of EphA receptors and ephrin-A ligands in motor neurons and limb mesenchymal cells. PMID- 12765611 TI - Filopodial calcium transients regulate growth cone motility and guidance through local activation of calpain. AB - Spontaneous intracellular calcium ([Ca2+](i)) transients in growth cone filopodia reduce filopodial motility, slow neurite outgrowth, and promote turning when generated asymmetrically; however, the downstream effectors of these Ca2+ dependent behaviors are unknown. We report that Ca2+ transients in filopodia activate the intracellular protease calpain, which slows neurite outgrowth and promotes repulsive growth cone turning upon local activation. Active calpain alters the balance between tyrosine kinase and phosphatase activities in filopodia, resulting in a net decrease in tyrosine phosphorylation, which mediates both filopodial stabilization and reduced lamellipodial protrusion. Our findings indicate that locally generated Ca2+ signals repel axon outgrowth through calpain-dependent regulation of phosphotyrosine signaling at integrin mediated adhesion sites. PMID- 12765612 TI - Activation of PI3-kinase is required for AMPA receptor insertion during LTP of mEPSCs in cultured hippocampal neurons. AB - Hippocampal CA1 homosynaptic long-term potentiation (LTP) is expressed specifically at activated synapses. Increased insertion of postsynaptic alpha amino-3-hydroxy-5-methyl-isoxazole-4-propionic acid receptors (AMPARs) appears to be crucial for CA1 LTP. However, the mechanism underlying AMPAR insertion during LTP remains largely unknown. We now report that phosphatidylinositol 3-kinase (PI3K) is complexed with AMPARs at synapses and activated by selective stimulation of synaptic N-methyl-D-aspartate (NMDA) receptors. Activation of the AMPAR-associated PI3K is required for the increased cell surface expression of AMPARs and LTP. Thus, our results strongly suggest that the AMPAR-PI3K complex may constitute a critical molecular signal responsible for AMPAR insertion at activated CA1 synapses during LTP, and consequently, this lipid kinase may serve to determine the polarity of NMDA receptor-dependent synaptic plasticity. PMID- 12765613 TI - Neural encoding in ventral striatum during olfactory discrimination learning. AB - A growing body of evidence implicates the ventral striatum in using information acquired through associative learning. The present study examined the activity of ventral striatal neurons in awake, behaving rats during go/no-go odor discrimination learning and reversal. Many neurons fired selectively to odor cues predictive of either appetitive (sucrose) or aversive (quinine) outcomes. Few neurons were selective when first exposed to the odors, but many acquired this differential activity as rats learned the significance of the cues. A substantial proportion of these neurons encoded the cues' learned motivational significance, and these neurons tended to reverse their firing selectivity after reversal of odor-outcome contingencies. Other neurons that became selectively activated during learning did not reverse, but instead appeared to encode specific combinations of cues and associated motor responses. The results support a role for ventral striatum in using the learned significance, both appetitive and aversive, of predictive cues to guide behavior. PMID- 12765614 TI - Effects of stimulus-response compatibility on neural selection in frontal eye field. AB - We investigated the neural basis of visual and saccade selection in the frontal eye field of macaque monkeys using a singleton search task with prosaccade or antisaccade responses. Two types of neurons were distinguished. The first initially selected the singleton even in antisaccade trials, although most subsequently selected the endpoint of the saccade. The time the singleton was located was not affected by stimulus-response compatibility and did not vary with reaction time across trials. The second type of neuron selected only the endpoint of the saccade. The time of endpoint selection by these neurons accounted for most of the effect of stimulus-response compatibility on reaction time. These results indicate that visual selection and saccade selection are different processes. PMID- 12765615 TI - Correlated neuronal discharges that increase coding efficiency during perceptual discrimination. AB - During a sensory discrimination task, the responses of multiple sensory neurons must be combined to generate a choice. The optimal combination of responses is determined both by their dependence on the sensory stimulus and by their cofluctuations across trials-that is, the noise correlations. Positively correlated noise is considered deleterious, because it limits the coding accuracy of populations of similarly tuned neurons. However, positively correlated fluctuations between differently tuned neurons actually increase coding accuracy, because they allow the common noise to be subtracted without signal loss. This is demonstrated with data recorded from the secondary somatosensory cortex of monkeys performing a vibrotactile discrimination task. The results indicate that positive correlations are not always harmful and may be exploited by cortical networks to enhance the neural representation of features to be discriminated. PMID- 12765616 TI - Cortical magnification within human primary visual cortex correlates with acuity thresholds. AB - We measured linear cortical magnification factors in V1 with fMRI, and we measured visual acuity (Vernier and grating) in the same observers. The cortical representation of both Vernier and grating acuity thresholds in V1 was found to be roughly constant across all eccentricities. We also found a within-observer correlation between cortical magnification and Vernier acuity, further supporting claims that Vernier acuity is limited by cortical magnification in V1. PMID- 12765617 TI - Patterns of mandibular variation in Pan and Gorilla and implications for African ape taxonomy. AB - Pan and Gorilla taxonomy is currently in a state of flux, with the number of existing species and subspecies of common chimpanzee and gorilla having been recently challenged. While Pan and Gorilla systematics have been evaluated on the basis of craniometric and odontometric data, only a handful of studies have evaluated multivariate craniometric variation within P. troglodytes, and none have evaluated in detail mandibular variation in either P. troglodytes or Gorilla gorilla. In this paper, we examine ontogenetic and adult mandibular variation in Pan and Gorilla. We test the hypothesis that patterns and degrees of mandibular variation in Pan and Gorilla closely correspond to those derived from previous analyses of craniometric variation. We then use these data to address some current issues surrounding Pan and Gorilla taxonomy. Specifically, we evaluate the purported distinctiveness of P.t. verus from the other two subspecies of Pan troglodytes, and the recent proposals to recognize Nigerian gorillas as a distinct subspecies, Gorilla gorilla diehli, and to acknowledge mountain and lowland gorillas as two separate species. Overall, patterns and degrees of multivariate mandibular differentiation parallel those obtained previously for the cranium and dentition. Thus, differences among the three conventionally recognized gorilla subspecies are somewhat greater than among subspecies of common chimpanzees, but differences between P. paniscus and P. troglodytes are greater than those observed between any gorilla subspecies. In this regard, the mandible does not appear to be more variable, or of less taxonomic value, than the face and other parts of the cranium. There are, however, some finer differences in the pattern and degree of morphological differentiation in Pan and Gorilla, both with respect to cranial and dental morphology, and in terms of the application and manner of size adjustment. Mandibular differentiation supports the conventional separation of bonobos from chimpanzees regardless of size adjustment, but size correction alters the relative alignment of taxa. Following size correction, intergroup distances are greatest between P.t. verus and all other groups, but there is considerable overlap amongst chimpanzee subspecies. Amongst gorillas, the greatest separation is between eastern and western gorillas, but adjustment relative to palatal vs. basicranial length results in a greater accuracy of group classification for G.g. gorilla and G.g. graueri, and more equivalent intergroup distances amongst all gorilla groups. We find no multivariate differentiation of the Nigerian gorillas based on mandibular morphology, suggesting that the primary difference between Nigerian and other western lowland gorillas lies in the nuchal region. Though intergroup distances are greatest between P.t. verus and other chimpanzee subspecies, the degree of overlap amongst all three groups does not indicate a markedly greater degree of distinction in mandibular, as opposed to other morphologies. Finally, mandibular differentiation corroborates previous craniodental studies indicating the greatest distinction amongst gorillas is between eastern and western groups. Thus, patterns and degrees of mandibular variation are in agreement with other kinds of data that have been used to diagnose eastern and western gorillas as separate species. PMID- 12765618 TI - Energy transformation during erect and 'bent-hip, bent-knee' walking by humans with implications for the evolution of bipedalism. AB - We have previously reported that predictive dynamic modeling suggests that the 'bent-hip, bent-knee' gait, which some attribute to Australopithecus afarensis AL 288-1, would have been much more expensive in mechanical terms for this hominid than an upright gait. Normal walking by modern adult humans owes much of its efficiency to conservation of energy by transformation between its potential and kinetic states. These findings suggest the question if, and to what extent, energy transformation exists in 'bent-hip, bent-knee' gait. This study calculates energy transformation in humans walking upright, at three different speeds, and walking 'bent-hip, bent-knee'. Kinematic data were gathered from video sequences and kinetic (ground reaction force) data from synchronous forceplate measurement. Applying Newtonian mechanics to our experimental data, the fluctuations of kinetic and potential energy in the body centre of mass were obtained and the effects of energy transformation evaluated and compared. In erect walking the fluctuations of two forms of energy are indeed largely out-of-phase, so that energy transformation occurs and total energy is conserved. In 'bent-hip, bent knee' walking, however, the fluctuations of the kinetic and potential energy are much more in-phase, so that energy transformation occurs to a much lesser extent. Among all modes of walking the highest energy recovery is obtained in subjectively 'comfortable' walking, the next highest in subjectively 'fast' or 'slow' walking, and the least lowest in 'bent-hip, bent-knee' walking. The results imply that if 'bent-hip, bent-knee' gait was indeed habitually practiced by early bipedal hominids, a very substantial (and in our view as yet unidentified) selective advantage would have had to accrue, to offset the selective disadvantages of 'bent-hip, bent-knee' gait in terms of energy transformation. PMID- 12765619 TI - The carbon isotope ecology and diet of Australopithecus africanus at Sterkfontein, South Africa. AB - The stable carbon isotope ratio of fossil tooth enamel carbonate is determined by the photosynthetic systems of plants at the base of the animal's foodweb. In subtropical Africa, grasses and many sedges have C(4)photosynthesis and transmit their characteristically enriched 13C/(12)C ratios (more positive delta13C values) along the foodchain to consumers. We report here a carbon isotope study of ten specimens of Australopithecus africanus from Member 4, Sterkfontein (ca. 2.5 to 2.0Ma), compared with other fossil mammals from the same deposit. This is the most extensive isotopic study of an early hominin species that has been achieved so far. The results show that this hominin was intensively engaged with the savanna foodweb and that the dietary variation between individuals was more pronounced than for any other early hominin or non-human primate species on record. Suggestions that more than one species have been incuded in this taxon are not supported by the isotopic evidence. We conclude that Australopithecus africanus was highly opportunistic and adaptable in its feeding habits. PMID- 12765620 TI - Optical dating of dune sand from Blombos Cave, South Africa: I--multiple grain data. AB - An aeolian sand unit overlies the Middle Stone Age deposits at Blombos Cave on the southern Cape coast. These deposits contained culturally-important artefacts, including bone tools and pieces of engraved ochre, as well as a large number of worked lithics. The aeolian sand and two other remnants of the sand dune formed against the coastal cliff were dated using optical dating. To determine the dose received since deposition, measurements were made on 5mg aliquots of purified quartz grains using the single-aliquot regenerative-dose (SAR) protocol. The results of several internal check procedures are reported and at least 15 replicate dose determinations are presented for each sample. Combining these dose values with measurements of the radioactive content of each sample resulted in an age of 69.2+/-3.9 ka for the unit within the cave, and a mean age of 70.1+/-1.9 ka for all three dune samples. This provides a minimum age for the Middle Stone Age material at Blombos Cave. PMID- 12765621 TI - Optical dating of dune sand from Blombos Cave, South Africa: II--single grain data. AB - A sequence of optically stimulated luminescence measurements was made on each of 8,961 grains from three sand samples from Blombos on the southern Cape coast. One sand unit overlay Middle Stone Age deposits in Blombos Cave. The measurement sequence, the single aliquot regenerative dose protocol, was used to obtain values for the total effective radiation dose to which each grain had been exposed since burial. A series of checks was carried out on each grain to ensure that the luminescence signals were reproducible, and that they were derived from quartz. This led to acceptance of less than 5% of the grains. An estimate of the radiation dose for the sand unit was obtained by combining the values using the central age model. In order to use a larger number of grains that might be representative of the sand unit, the radiation dose was also estimated by using the signal from the above grains, combined with the signals from those grains that had lower signals, but nonetheless contributed to the total light sum; this utilised between 9 and 18% of the grains. This enables us to obtain estimates of the ages as 67.3+/-3.8 ka, 65.6+/-2.8 ka and 68.8+/-3.0 ka for the three samples. These values agree with ages obtained using the single aliquot regenerative dose protocol for aliquots composed of several hundred grains. PMID- 12765622 TI - The phylogenetic relationship of Neandertal and modern human mitochondrial DNAs based on informative nucleotide sites. PMID- 12765623 TI - Magnetostratigraphy of the Evron Member--implications for the age of the Middle Acheulian site of Evron Quarry. PMID- 12765625 TI - Feline idiopathic cardiomyopathy: a retrospective study of 106 cats (1994-2001). AB - The case records of 106 cats with idiopathic cardiomyopathy that presented to the Feline Centre of the University of Bristol between September 1994 and September 2001 were reviewed retrospectively. Hypertrophic cardiomyopathy (HCM) was the most common form seen (57.5%), followed by restrictive cardiomyopathy (RCM) (20.7%), dilated cardiomyopathy (DCM) (10.4%) and unclassified cardiomyopathy (UCM) (10.4%). One cat showed echocardiographic changes compatible with a moderator band cardiomyopathy (MBCM). Most affected cats were domestic short hairs (DSH) (57.5%). The mean (+/-SD, range) age of cats with cardiomyopathy at presentation was 6.8 (4.3, 0.5-16) years, with an equal distribution of males and females. Clinical findings, electrocardiographic changes and radiographic abnormalities were also reviewed. The median survival time for 73 cats for which follow-up data was available was 300 days. A greater survival time was observed for cats with UCM (925 days) when compared with those with HCM (492 days), RCM (132 days) or DCM (11 days). PMID- 12765626 TI - The prevalence of immediate and delayed type hypersensitivity reactions to Microsporum canis antigens in cats. AB - Spontaneous recovery from Microsporum canis infections in cats is thought to be dependent on the development of a competent immune response. The purpose of this study was to determine the prevalence of positive delayed type hypersensitivity reactions in cats with and without dermatophytosis. Four groups of cats were intradermally skin tested with M canis extract and test sites were evaluated both subjectively and objectively at 0, 24 and 48 h after injection. Delayed intradermal testing (IDT) reactions were absent in cats not exposed to dermatophytosis (n=20); infected-recovered cats (n=38 culture negative lesion negative and n=43 lesion negative but culture positive) had significantly larger IDT reactions than unexposed cats and cats that were still actively infected (n=18). Based on the results of this study, IDT with M canis extract can be used to assess the cellular immune response of cats with dermatophytosis. PMID- 12765627 TI - Short chain fatty acids stimulate feline colonic smooth muscle contraction. AB - The effect of short chain fatty acids (SCFA) on feline colonic smooth muscle contraction was evaluated in vitro. Colonic tissue was obtained from seven healthy male and female adult cats and seven healthy male and female kittens. Longitudinal and circular colonic smooth muscle strips from proximal and distal colon were incubated with SCFA (acetate, butyrate and propionate; 1-100mM). SCFA induced contractions were compared to responses obtained using maximal concentrations (10(-4)M) of acetylcholine (ACh). The calcium dependence of the SCFA response was investigated by incubating with nifedipine (1 microM) or verapamil (1 microM). Acetate, butyrate and propionate elicited isometric stress responses (0.25-1.98 x 10(4)N/m(2)) in longitudinal, but not circular, smooth muscle from both the proximal and distal colon of adult cats. Maximal responses were attained at 50 and 100mM SCFA. Maximal butyrate and propionate responses were 29 and 19% of the maximal ACh response (10(-4)M), respectively. Acetate was least effective in stimulating contractile responses. Nifedipine and verapamil abolished all responses. Contractile responses in kittens were similar to those observed in adult cats, but were smaller in amplitude. Results of these studies have shown that SCFA stimulate longitudinal colonic smooth muscle contractions in kittens and adult cats in vitro. These SCFA-induced contractions involve activation of calcium influx. These in vitro findings may account for some of the effects of dietary fiber on feline colonic motility in vivo. PMID- 12765628 TI - Single-injection inulin clearance for routine measurement of glomerular filtration rate in cats. AB - Glomerular filtration rate (GFR) was determined in 53 cats using an inulin single injection method. Thirty healthy young adult cats were used to establish normal values. The procedure was also used in 23 cats that were either older than 10 years or had borderline serum creatinine levels. The total clearance was calculated from the decay of the serum inulin concentration after injection of 3000 mg/m(2)body surface area using a two-compartment model. Concomitant inulin and iohexol clearance in nine cats showed excellent correlation between the two methods. Calculated normal values for GFR in 30 healthy cats were 35.9-58.5 (median 46.0) ml/min/m(2)or 2.07-3.69 (median 2.72) ml/min/kg. A few cats with normal creatinine or blood urea nitrogen levels were detected as having reduced GFR and therefore being in a state of early renal dysfunction. The study indicates that single-injection inulin clearance is a valuable tool for routine GFR measurement in cats. An "inulin excretion test" using only one blood sample 3h after the administration of 3000 mg/m(2)body surface area could prove an attractive alternative for the assessment of renal function in daily practice. PMID- 12765629 TI - Treatment of feline diabetes mellitus using an alpha-glucosidase inhibitor and a low-carbohydrate diet. AB - The purpose of this study was to determine the effect of an alpha-glucosidase inhibitor (acarbose), combined with a low-carbohydrate diet on the treatment of naturally occurring diabetes mellitus in cats. Eighteen client-owned cats with naturally occurring diabetes mellitus were entered into the study. Dual-energy X ray absorptiometry (DEXA) was performed prior to and 4 months after feeding the diet to determine total body composition, including lean body mass (LBM) and percent body fat. Each cat was fed a commercially available low-carbohydrate canned feline diet and received 12.5mg/cat acarbose orally every 12h with meals. All cats received subcutaneous insulin therapy except one cat in the study group that received glipizide (5mg BID PO). Monthly serum glucose and fructosamine concentrations were obtained, and were used to adjust insulin doses based on individual cat's requirements. Patients were later classified as responders (insulin was discontinued, n=11) and non-responders (continued to require insulin or glipizide, n=7). Responders were initially obese (>28% body fat) and non responders had significantly less body fat than responders (<28% body fat). Serum fructosamine and glucose concentrations decreased significantly in both responder and non-responder groups over the course of 4 months of therapy. Better results were observed in responder cats, for which exogenous insulin therapy was discontinued, glycemic parameters improved, and body fat decreased. In non responders, median insulin requirements decreased and glycemic parameters improved significantly, despite continued insulin dependence. The use of a low carbohydrate diet with acarbose was an effective means of decreasing exogenous insulin dependence and improving glycemic control in a series of client-owned cats with naturally occurring diabetes mellitus. PMID- 12765631 TI - Diaphragmatic hernia in a cat mimicking a pulmonary mass. AB - A seven-year-old castrated British shorthair cross cat was presented for coughing of five-weeks duration. Thoracic radiographs and an unguided bronchoalveolar lavage showed changes consistent with inflammatory airway disease. In addition, a soft tissue density was evident in the thoracic films between the heart and the diaphragm. Exploratory thoracotomy demonstrated a diaphragmatic hernia, probably congenital in origin, with incarceration of a portion of the hepatic parenchyma. The herniated portion of liver was resected surgically and the defect in the diaphragm closed. The cat was given a 10-day course of doxycycline post operatively and the cough did not recur subsequently. In retrospect, the hernia was potentially an incidental problem, the cat's coughing being attributable to inflammatory airway disease. PMID- 12765630 TI - The short-term clinical efficacy of amitriptyline in the management of idiopathic feline lower urinary tract disease: a controlled clinical study. AB - In a controlled study, the effects of amitriptyline compared with that of a placebo in cats suffering from idiopathic Feline Lower Urinary Tract Disease (FLUTD) have been investigated. Thirty-six animals were selected by veterinary practitioners and treated with a placebo or 10mg amitriptyline once daily. All animals received concomitant antibiotic treatment. A total of 24 cats were included in the final assessment of the results. The severity of symptoms before and after treatment were compared between groups and showed no significant difference. Results indicated that the 7-day course of 10mg amitriptyline was not effective in the treatment of idiopathic FLUTD. Thus, it is considered not to be beneficial as a short-term therapy where the therapeutic results depend on peripheral effects of the drug. Long-term effects may be expected 4 or more weeks after the start of therapy and need to be further investigated. PMID- 12765632 TI - Power and conflict in intensive care clinical decision making. AB - It is clear that current government policy places increasing emphasis on the need for flexible team working. This requires a shared understanding of roles and working practices. However, review of the current literature reveals that such a collaborative working environment has not as yet, been fully achieved. Role definitions and power bases based on traditional and historical boundaries continue to exist. This ethnographic study explores decision making between doctors and nurses in the intensive care environment in order to examine contemporary clinical roles in this clinical specialty. Three intensive care units were selected as field sites and data was collected through participant observation, ethnographic interviews and documentation. A key issue arising in this study is that whilst the nursing role in intensive care has changed, this has had little impact on how clinical decisions are made. Both medical and nursing staff identify conflict during patient management discussions. However, it is predominantly nurses who seek to redress this conflict area through developing specific behaviours for this clinical forum. Using this approach to resolve such team issues has grave implications if the government vision of interdisciplinary team working is to be realised. PMID- 12765633 TI - Developing a competency framework for critical care to match patient need. AB - The competency framework developed by the critical care education group of the London Standing Conference aims to serve every grade and level of practitioner. It is neither time specific nor static. The patient is the central focus of the framework and the elements of competence reflect patient need at any critical care level [Comprehensive Critical Care: A Review of Adult Critical Care Services, The Stationary Office, London]. A group of expert nurses have developed the competency framework, with widespread consultation and collaboration. This approach intended to develop consistency for critical care education and practice. It is envisaged that this will reduce pockets of repeated activity, which places huge demands on limited resources. The critical care competency framework was developed using the method of functional analysis. A plan for the future has been identified, including continued collaboration and consultation with Trusts and Higher Educational Institutions and the development of an online manual to support the competency framework. KEY POINTS: 1. Critical care delivery has been under close scrutiny and a number of key contemporary drivers have led to the development of this competency framework. 2. The development of a consistent pan-London approach to critical care education has been identified. 3. The patient is the focus of critical care delivery and therefore patient need is central to the critical care competency framework. 4. Wider collaboration is needed with other agencies and groups to prevent the repetition of work already carried out. PMID- 12765634 TI - Respiratory care in spinal cord injury with associated traumatic brain injury: bridging the gap in critical care nursing interventions. AB - Spinal cord injury (SCI) is a devastating and challenging condition. The events that lead to SCI, such as road traffic accidents, falls, sports and violence [Top. Spinal Cord Inj. Rehabil. 5 (1999) 83], are also the common aetiologies of traumatic brain injury (TBI). It's not surprising then, that 20-50% of those with cervical SCI have TBI [J. Trauma 46 (1999) 450]. The literature pertaining to the management of either injury in isolation is vast, but lacking where the two conditions are experienced together and require distinct adaptations to interventions. Consequently, a gap in the literature exists. This paper focuses on those patients with SCI of the cervical spine with associated head injury, and pay particular attention to respiratory difficulties, and presents interventions required to minimise and treat the effects of such pulmonary compromise. PMID- 12765636 TI - Pressure sore prevention in the critically ill: what you don't know, what you should know and why it's important. AB - The critically ill are particularly vulnerable to pressure sore development. These expensive and often painful complications have been largely ignored for many years and the entire problem has been managed by nursing staff. Current methods for identifying patients at risk are inadequate and subjective. Scoring systems have been known to over-predict those at risk and this maybe because they frequently originate from elderly care settings. Additionally, their relevance to the critically ill has not yet been established. The use of pressure-relieving devices has become commonplace; however, there is a paucity of data from controlled clinical studies. No uniform approach in measuring the effectiveness of these devices exists. What is certain, though, is that a voluminous amount of work needs to be conducted in order to verify their continued use. It is increasingly apparent that the complex nature of pressure sore development means that it is unrealistic to expect a single discipline to manage the problem effectively. A multidisciplinary team approach is the most appropriate way to improve management in this vital area. PMID- 12765635 TI - Pain assessment in critical care: what have we learnt from research. AB - Despite an ongoing acknowledgement in the literature that pain is a significant problem within the critical care environment, this issue has not been adequately addressed by critical care nurses. This paper examines strategies for changing pain management practices in critical care, including reviewing documentation practices, the utilisation of guidelines and algorithms to augment clinical decision making, and increasing educational opportunities available to critical care nurses. It is recommended that pain assessment be given a higher priority within the clinical context, particularly as inadequate pain assessment and management has been linked to increased morbidity and mortality within critical care. Importantly, critical care nurses need to not only be aware of research based pain management practices, but also lead the way in implementation and continuous evaluation as a measure of decreasing patient pain in the future. PMID- 12765637 TI - Optimisation of oxygenation and tissue perfusion in surgical patients. AB - Surgical patients with limited cardiovascular reserve have much worse prognosis than patients with normal hearts. This review identifies 17 randomised controlled clinical trials that have investigated peri-operative therapy designed to increase tissue perfusion in surgical patients, many of whom have limited cardiovascular reserve. Although there are differences which make equating the trials complex, a total of 1974 patients have been enrolled in the studies and the odds ratio for reduction in mortality is 0.45 (95% confidence intervals 0.33 0.60). Further research needs to be undertaken in the identification of patients with limited cardiovascular reserve and for investigating proposed treatment strategies. Despite this, it appears that such patients have improved outcome if they are admitted to intensive care unit pre-operatively and have suitable therapy given to improve tissue oxygen delivery. PMID- 12765638 TI - The use of peptidomics in endocrine research. AB - In 2002, the Nobel Prize for chemistry was awarded to the inventors of two novel ionization techniques in mass spectrometry, MALDI and ESI. These techniques, often in combination with data from genomic databases, represent an extremely powerful tool in analytical (bio)chemistry, with many applications, e.g., in the field of proteomics. Peptides, which are small proteins, have, despite their importance as controlling agents in numerous physiological processes, as yet been much less intensively studied by these novel techniques than larger proteins. The term peptidomics, i.e., the study of all peptides expressed by a certain cell, organ or organism was only introduced in 2001. In neuroendocrinology, spectacular progress could already be realized and the future looks bright. In this minireview we discuss the different methodologies that are used in peptidomics and give an overview of the wide range of applications. PMID- 12765639 TI - Mode of action of neuropeptides from the adipokinetic hormone family. AB - Neuropeptides of the adipokinetic hormone (AKH) family regulate inter alia mobilisation of various substrates from stores in the fat body of insects during episodes of flight. How is this achieved? In insects which exclusively oxidise carbohydrates for flight (cockroaches), or which oxidise carbohydrates in conjunction with lipids (locusts) or proline (a number of beetles), the endogenous AKHs bind to a G(q)-protein-coupled receptor, activate a phospholipase C and the resulting inositol trisphosphate releases Ca(2+) from internal stores. In addition, influx of extracellular Ca(2+) is increased and, via a kinase cascade, glycogen phosphorylase is activated, glucose-1-phosphate produced, and transformed to trehalose, which is released into the haemolymph. In locusts, additionally, adenylate cyclase is activated and cyclic AMP is synthesised. In insects which use lipids for sustained flight (locust, tobacco hornworm moth) or proline for flight (certain beetles), adenylate cyclase is activated after the AKHs bind to their respective G(s)-protein-coupled receptor. The resulting cyclic AMP, together with the messengers intra- and extracellular Ca(2+), activate a triacylglycerol lipase, which results in the production of 1,2 diacylglycerols (in locusts, moths) or (hypothetically) free fatty acids (fruit beetle). PMID- 12765640 TI - Within-sample variation of fecal glucocorticoid measurements. AB - Fecal glucocorticoid metabolite analysis is a useful tool for monitoring adrenocortical activity in captive and free-ranging wildlife. Glucocorticoid metabolites may not be evenly distributed within fecal samples and this variability could affect the interpretation of glucocorticoid metabolite measurements. Furthermore, the precision (i.e., repeatability of measurements) of fecal glucocorticoid measurements from well-mixed samples is unknown. We collected fresh white-tailed deer (Odocoileus virginianus) feces at various times pre- and post-adrenocorticotropin injection to provide samples with low (<75 ng/g), medium (75-90 ng/g), and high (>90 ng/g) glucocorticoid concentrations in case variability differs in samples of dissimilar hormone metabolite concentrations. We compared two sampling methods (selection of three pellet groups [one from each end of the fecal mass and one from the center] versus sampling three small portions of the thoroughly mixed fecal mass) to estimate within-sample variation of glucocorticoid metabolites. Glucocorticoid metabolite measures from pellet groups were higher than fecal glucocorticoid measures from mixed samples in the low (F=3.10; df = 1,56; P=0.08) and medium concentration (F=7.28; df = 1,50; P=0.01) groups. Fecal glucocorticoid metabolite estimates from mixed samples were less variable than glucocorticoid metabolite measures using pellet groups from the same fecal mass, although these differences were not statistically significant (low group: F=0.59; df = 1,38; P=0.45; medium group: F=0.13; df = 1,34; P=0.72; high group: F=2.30; df = 1,28; P=0.14). The mean coefficient of variation was <10% across all treatment groups and sampling methods. However, a power analysis indicated the mixed sub-sample technique requires fewer samples to detect statistically significant differences than pellet groups. Our results suggest that glucocorticoid metabolites may not be evenly distributed in white-tailed deer feces. Consequently, using only a few pellets from a fecal mass may bias assay interpretation. We suggest researchers homogenize the entire fecal mass before removing a sub-sample of fecal material for analysis. Also, other sources of variation must be considered when interpreting results of fecal glucocorticoid studies. PMID- 12765641 TI - Serum relaxin concentrations and reproduction in male bonnethead sharks, Sphyrna tiburo. AB - Relaxin is a 6-kd polypeptide hormone that is responsible for regulating several reproductive processes in female vertebrates, but its role in male reproduction remains unclear. To aid in clarifying this role, the objective of the present study was to investigate changes in endogenous relaxin levels associated with reproductive events in male elasmobranchs, which represent one of only three vertebrate groups known to possess this hormone. Serum relaxin concentrations were measured in 27 immature and 66 mature male bonnethead sharks (Sphyrna tiburo), a species with a well-characterized, seasonal reproductive cycle. Temporal changes in serum relaxin concentrations of immature male S. tiburo were not observed. In contrast, a temporal cycle in serum relaxin concentrations of mature male S. tiburo was observed in individuals from two sampling locations. Significant increases (P<0.05) in serum relaxin concentrations of mature male S. tiburo from both collection sites occurred during late spermatogenesis and the mating period, two critical stages of the reproductive cycle. The results from this study suggest that relaxin may play an important role in regulating semen quality, or other aspects of reproduction in male sharks. This is the first study to demonstrate a temporal pattern in endogenous serum Rlx concentrations associated with reproductive events in feral vertebrates. As such, it strengthens earlier hypotheses that suggested a role for this hormone in regulating male vertebrate fertility and copulatory success. PMID- 12765642 TI - Immunocytochemical applications of specific antisera raised against synthetic fragment peptides of mummichog GtH subunits: examining seasonal variations of gonadotrophs (FSH cells and LH cells) in the mummichog and applications to other acanthopterygian fishes. AB - Two distinct types of gonadotrophs, FSH (GtH I) cells and LH (GtH II) cells, were immunocytochemically identified from mummichog (Fundulus heteroclitus; Cyprinodontiformes, Acanthopterygii) pituitary using antisera raised against synthetic fragment peptides of FSHbeta and LHbeta. Both cell types were abundant during the spawning period (spring and early summer) and decreased in number during the post-spawning immature period. The number of FSH cells increased again during the early phases of gonadal development (cortical alveoli accumulation in the oocytes and basal spermatogenesis) in early winter, whereas the number of LH cells did not. Only FSH cells were abundant during the latter phases of gonadal development (vitellogenesis and active spermatogenesis) in early spring. These observations suggest that both GtHs have important yet different roles for reproduction in this species. Antisera against the conservative region of the FSHbeta and the LHbeta subunits immunostained FSH cells and LH cells, respectively, also in red seabream (Pagrus major; Perciformes, Acanthopterygii) and small mouth bass (Micropterus dolomieu; Perciformes, Acanthopterygii), suggesting the possibility of their use for other acanthopterygian fishes. PMID- 12765643 TI - A novel function of cockroach (Periplaneta americana) hypertrehalosemic hormone: translocation of lipid from hemolymph to fat body. AB - The hypertrehalosemic hormones Pea HTH-I and Pea HTH-II have a lipogenic effect in the cockroach, Periplaneta americana. This has been determined by measuring the change in tri-, di-, monoacylglycerol, phospholipid and free fatty acids in the hemolymph and fat body in response to the hormones. The effect of both hormones 6h after injection is comparable and is expressed as an increase in fat body triacylglycerol and a decrease in hemolymph triacylglycerol. Coincident with this effect the concentration of diacylglycerol in the hemolymph decreases while that in the fat body increases. Hemolymph and fat body phospholipid are both lowered by HTH. The concentration of free fatty acids in the hemolymph, as well as fat body, are increased by both hormones. Confirmation of the lipogenic effect of HTH has been obtained by feeding cockroaches [3H]oleic acid and measuring the incorporation of label into hemolymph and fat body lipid. The data show that the specific activity of the hemolymph phospholipid fraction increases more than 50%. In fat body, the specific activity of the label in the triacylglycerol fraction increased more than 2-fold while that in the free fatty acid fraction increased more than 20-fold. The data support the view that the fatty acids incorporated into fat body triacylglycerol under the influence of HTH are not formed de novo but are derived from the sn-1 and sn-2 positions of phospholipid which is initially formed in the midgut wall. PMID- 12765644 TI - Luteal development and progesterone levels during pregnancy of the viviparous temperate lizard Barisia imbricata imbricata (Reptilia: Anguidae). AB - The relationship between plasma progesterone (P(4)) levels and the formation and degeneration of the corpus luteum (CL) was assessed monthly during gestation of the viviparous lizard Barisia imbricata imbricata. Histochemical activity of the delta(5-4) isomerase 3 beta-hydroxysteroide dehydrogenase (delta(5-4)3beta-HSD) in the luteal tissue and embryonic development were also observed. Females were gravid throughout winter and great part of spring (late November or early December until late May or early June). Corpus luteum development occurred in the first third of gestation (December and January) when the embryo reached developmental stage 27. Four sequential stages were identified during development and three stages during regression of the CL. The follicular and thecal tissue participated in the formation of the luteal cell mass. According to Xavier's classification, the CL of B. i. imbricata is a subtype from Type III. The activity of delta(5-4)3beta-HSD was observed mainly in the luteal cell mass. The first degenerative changes in the CL were observed in the early second third of the gestation and continued gradually until parturition. Progesterone levels increased in early pregnancy and reached its highest level during January (3.07+/ 1.04 ng/ml) when mature corpora lutea were present. Gradual diminution in progesterone concentrations occurred in the second and last third of pregnancy and coincided with advanced degenerative changes and diminution in histochemical activity of delta(5-4)3beta-HSD in the luteal tissue. These observations suggest that the CL is the major source of progesterone during pregnancy of B. i. imbricata. PMID- 12765645 TI - Localization and role of serotonin in the adrenal gland of Podarcis sicula (Reptilia, Lacertidae). AB - The occurrence of serotonin (5-hydroxytryptamine; 5-HT) in the chromaffin cells of Podarcis sicula adrenal gland was demonstrated by immunocytochemical techniques: ABC and immunogold methods. At LM and EM levels, antiserum against 5 HT revealed serotonin immunoreactivity prevalently in noradrenalin (NA) cells, on and around secretory vesicles; adrenalin (A) cells appeared scarcely stained. The role of serotonin in the regulation of adrenal gland activity was studied in vivo using LM and EM techniques coupled to a specific radioimmunoassay for adrenocorticotropic hormone (ACTH) and corticosterone. 5-HT (0.7 mg/100 g body wt)/day for 4 days increased ACTH and corticosterone release; at LM and EM level clear signs of stimulation in the steroidogenic tissue were observed, as evidenced by the variations of lipid/cytoplasm ratio. In the chromaffin tissue, LM observations evidenced a variation of the numeric NA/A cell ratio; at EM level, chromaffin tissue showed intermediate cells with A, NA, and very clear granules with granular elements. The occurrence of these cells might be the result of a process of resynthesis following serotonin-stimulated catecholamine release. These data suggested that serotonin might be involved in the modulation of Podarcis pituitary-adrenal axis, and act as a paracrine factor to modulate corticosteroid production. PMID- 12765646 TI - Differential expression and cellular localization of somatolactin-1 and -2 during early development in the gilthead sea bream. AB - The patterns of expression of the somatolactin 1 and 2 (SL1 and SL2) transcripts were studied during the early development of the gilthead sea bream (Sparus aurata). Gene expression of SL1 and SL2 were detected in embryos and in larvae, although both transcripts presented different levels of expression. The SL1 transcripts in contrast to the SL2 transcripts presented high expression levels in embryos and younger larvae. Moreover, the SL2 transcripts were slightly present or absence in embryonic stage and the newly hatched larvae, respectively. The differences in the expression levels of SL1 and SL2 in embryos and larvae may be due to the fact that two distinct genes express both isoforms of the protein. Thus, both SLs may play different physiological roles throughout development. Moreover, the hybridization signals for SL1- and SL2-mRNAs were detected in 4-day old larvae. Both in larvae and adults the somatolactotroph cells co-expressed both transcripts of SL and were located bordering the neurohypophysis in the pars intermedia. PMID- 12765647 TI - ACTH occurrence in teleosts supramedullary neuron clusters: a neuron-glial common language? AB - The cross-talk between neurons and glia is receiving increased attention because of its potential role in information processing in the nervous system. We choose the cluster of supramedullary neurons (SN) and glial cells of pufferfish as a suitable model for studying neuron-glial interactions, identifying the implicated cell types and the signalling involved. In particular, among proopiomelanocortin (POMC)-derived peptides, adrenocorticotrope hormone (ACTH)-immunopositivity was found both in SN and in microglial cells. The present results for the first time show the presence of ACTH in microglia of a vertebrate. The role of ACTH is discussed, including its possible neuroprotective function. Moreover, SN immunoreactivity supports the idea that ACTH participates in neurotransmission and/or neuromodulation. In addition to these possible functions, the hypothesis is put forward that ACTH represents a common language by which neurons and glial cells communicate with each other. PMID- 12765648 TI - Fushi tarazu factor-1 mRNA and protein is expressed in steroidogenic and cholesterol metabolising tissues during different life stages in Arctic char (Salvelinus alpinus). AB - Fushi tarazu factor-1 (FTZ-F1) genes belong to the nuclear receptor family 5A (NR5A). The distribution pattern of NR5A genes in teleosts suggests that they control functions separate to, or in addition to, those of other vertebrates. In mammals NR5A1 genes, including steroidogenic factor-1 (SF-1), are primarily involved in steroidogenesis. NR5A2 contain the alpha-fetoprotein transcription factor (FTF) genes, which protect mammalian embryos against maternal estrogens, and are involved in cholesterol transfer and metabolism. In this study we have analysed the expression of two Arctic char FTZ-F1 forms belonging to the NR5A2 group. The expression starts during early development and the transcripts are present in embryonic liver/pancreas and gonadal regions. The genes are up regulated during embryogenesis as the embryo develops towards hatch, as shown by increased mRNA and protein levels. In adult Arctic char the FTZ-F1 forms are primarily located to tissues involved in steroidogenesis as well as cholesterol metabolism. Thus, a division of NR5A into SF-1 (NR5A1) and FTF (NR5A2) specific functions does not appear to have occurred in teleosts. PMID- 12765649 TI - Purification of a 41-kDa insulin-like growth factor binding protein from serum of chinook salmon, Oncorhynchus tshawytscha. AB - In salmon, at least three insulin-like growth factor binding proteins (IGFBPs) with molecular masses of 41, 28, and 22kDa exist in serum. The 41-kDa IGFBP is up regulated by growth hormone treatment and down-regulated by fasting, suggesting that it is a homolog of IGFBP-3. We purified the 41-kDa IGFBP from chinook salmon serum by IGF-I affinity chromatography followed by reversed-phase high pressure liquid chromatography. Purified IGFBP appeared as doublet bands on electrophoresis and was N-glycosylated. Analysis of partial N-terminal amino acid sequence revealed that salmon 41-kDa IGFBP has the cysteine rich domain conserved among IGFBP family. In a binding assay using 125I-salmon IGF-I, purified 41-kDa IGFBP specifically bound salmon IGF-I, human IGF-I and human IGF-II, but neither Long R(3)IGF-I nor salmon insulin, showing that binding characteristics of the salmon IGFBP are similar to those of mammalian IGFBPs. Although the partial amino acid sequence of 41-kDa IGFBP showed highest homologies with zebrafish and seabream IGFBP-2, the highly conserved nature of the N-terminus makes it impossible to identify the type of IGFBP from partial sequence data. However, based on physiological responses, molecular weight and type of glycosylation, the 41-kDa IGFBP is most similar to mammalian IGFBP-3. PMID- 12765650 TI - Glucocorticoid receptor upregulation during seawater adaptation in a euryhaline teleost, the tilapia (Oreochromis mossambicus). AB - Cortisol is an important seawater (SW) osmoregulatory hormone in the Mozambique tilapia (Oreochromis mossambicus), a highly euryhaline cichlid able to live in environments ranging from fresh water (FW) to salinities well in excess of full strength seawater. Previous studies indicate that cortisol may promote SW adaptation by increasing gill chloride cell differentiation, Na(+)/K(+)-ATPase activity and subsequent excretion of excess salt following seawater acclimation. Despite cortisol's widely accepted role as a SW-adapting hormone, cortisol receptor regulation during SW acclimation is not well understood. The purpose of these studies was to determine whether the intracellular glucocorticoid receptor (GR) might be regulated in a manner consistent with cortisol's actions in SW adaptation. Saturation radioligand binding assays were conducted on gill cytoplasm preparations from fish sampled 4 and 24h and 4 and 14 days after transfer from FW to 2/3 SW or FW (control). Affinity (K(d)) of the gill GR remained constant over the timecourse, while numbers of receptors (B(max)) in SW fish were significantly elevated compared with controls at 24h and 4 days after transfer. Plasma osmolality was higher in fish transferred to SW for 24h, 4 days, and 14 days compared with those animals moved to FW. Plasma cortisol levels and hepatic cortisol binding remained constant between SW and FW fish throughout the timecourse of the salinity challenge. These studies indicate that seawater acclimation is accompanied by a specific upregulation of intracellular GR numbers in gill tissue. The lack of increase in circulating cortisol following SW adaptation may reflect enhancement of clearance of the steroid. It appears that an increase in cortisol receptors, which is closely associated with the rise in blood osmotic pressure that accompanies SW exposure, is an important component of cortisol's ability to promote SW adaptation in the tilapia. PMID- 12765651 TI - Estradiol reduces pituitary responsiveness to somatostatin (SRIF-14) and down regulates the expression of somatostatin sst2 receptors in female goldfish pituitary. AB - Sex steroid hormones have been shown to regulate somatostatin (SRIF) gene expression in goldfish brain, which in turn influences the regulation of GH secretion. In this study, the influences of sex steroids on pituitary responsiveness to SRIF-14 and the pituitary expression of a type two SRIF receptor (sst(2)) were examined. Results from in vitro perifusion of pituitary fragments show that pituitaries from estradiol-primed sexually regressed female fish have significantly lower GH release responsiveness to pulse exposure to SRIF 14 than pituitaries from control or testosterone-treated sexually regressed females. Results from in vitro static culture show that pituitaries from sexually mature female fish have lower GH release responsiveness to SRIF-14 than those from sexually regressed females. In addition, the sst(2) receptor mRNA levels in pituitaries from mature and recrudescent female fish are significantly lower than in sexually regressed female fish. Our results indicate that estradiol acts at the level of the pituitary to regulate GH secretion by influencing the responsiveness to SRIF-14. The underlying mechanism includes, in part, reduction of the expression of sst(2) receptors, presumably leading to the lower number of the receptors available for SRIF binding. PMID- 12765652 TI - An endocrinological update in toads: disparity between the cardiovascular effects of two angiotensin II analogs. AB - This study was undertaken to test the hypothesis that, in toads, the pressor effects of angiotensin II (ANG II) are partly due to the secondary effects of catecholamines. In Bufo marinus, blood pressure responses to bullfrog ([Val(5)]) ANG II administration were measured in animals pretreated (experimental group) and not pretreated (control group) with phentolamine, an alpha-adrenergic antagonist. At 1, 2, and 4 min following ANG II administration (50 ngkg(-1)), mean arterial pressure (MAP) of the experimental group was significantly less than in the control group (27.9+/-1.0, 28.2+/-1.6, and 25.8+/-2.1 mmHg versus 45.1+/-1.0, 39.4+/-2.8, and 35.5+/-3.6, respectively; n=6). Heart rate (f(H)) was unaffected by phentolamine. These data support our hypothesis. Previous authors, using human ([Ile(5)]) ANG II, have concluded that alpha-adrenergic mechanisms are not involved in the pressor effects of ANG II in B. marinus. In a separate group of toads, we examined MAP and f(H) responses to similar doses of human and bullfrog ANG II (100 ngkg(-1)). Bullfrog ANG II caused MAP to reach higher peak values (56.5+/-2.6 versus 37.2+/-0.4 mmHg; n=7) in less time (approximately 1 min versus approximately 8 min) than human ANG II. Furthermore, bullfrog ANG II caused f(H) to significantly increase from 41.3+/-5.1 to 79.3+/-2.3 beats min(-1) (at 1 min) whereas human ANG II caused no significant changes in f(H) throughout the measured time course. Higher doses of human ANG II (100 microgkg(-1)) invoked slow but large increases in MAP with non-significant decreases in f(H). These additional data suggest that the choice of exogenously administered hormone is an important one to consider in comparative endocrinological studies. PMID- 12765653 TI - Changes in androgen receptor mRNA expression in the forebrain and oviduct during the reproductive cycle of female leopard geckos, Eublepharis macularius. AB - Successful reproduction requires the coordination of reproductive physiology with behavior. The neural correlates of reproductive behavior have been elucidated in a variety of amphibians, mammals, and birds but relatively few studies have examined reptiles. Here we investigate differences in androgen receptor (AR) mRNA expression in the forebrain and oviduct between previtellogenic and late vitellogenic female leopard geckos, Eublepharis macularius. Plasma concentrations of testosterone (T) are low when females are previtellogenic and sexually unreceptive but increase dramatically during late vitellogenesis when females are receptive. In addition, receptivity can be induced by treatment with exogenous T. The relative abundance of AR-mRNA across various nuclei was greater in late vitellogenic than in previtellogenic females. This general pattern was observed in the medial preoptic area, anterior hypothalamus, external nucleus of the amygdala, dorsolateral aspect of the ventromedial hypothalamus, lateral septum, and periventricular hypothalamus. There were also clear differences in AR-mRNA expression among these nuclei. The pattern of gene expression observed in the brain was reversed within stromal cells of the oviduct where expression of AR mRNA decreased from the previtellogenic stage to the late vitellogenic stage. Overall, these data demonstrate that T concentration in the plasma, abundance of AR-mRNA in the brain and oviduct, and sexual behavior change coordinately during the reproductive cycle of female leopard geckos. Although the function of AR in the female leopard gecko is not yet clear, our results are in accord with growing evidence that androgens regulate numerous aspects of female physiology and behavior in vertebrates. PMID- 12765654 TI - Activin and transforming growth factor-beta as local regulators of ovarian steroidogenesis in the goldfish. AB - This study explores the hypothesis that activin and TGFbeta(1) serve as local regulators of ovarian function in the goldfish. Initial studies demonstrated the presence of TGFbeta in the ovary through the use RT-PCR, which amplified a 225 bp product from early vitellogenic (EVIT) and prematurational full-grown (PFG) follicles. This transcript showed high homology to TGFbeta in other teleosts. Both goldfish recombinant activin B and human recombinant TGFbeta(1) suppressed basal testosterone production by EVIT follicles incubated in vitro. Activin B also inhibited hCG-stimulated testosterone production by EVIT follicles. Our experiments suggest that activin B mediates these effects through actions at sites upstream of cholesterol formation and/or mobilization in the steroidogenic pathway, and through mechanisms that were independent of effects on cAMP formation. In experiments with PFG follicles, TGFbeta(1) decreased basal testosterone production. Activin B did not affect T production by PFG follicles, suggesting that this hormone has differential effects on steroidogenesis in the goldfish ovary depending on the stage of ovarian maturity. In other tests with PFG follicles, TGFbeta(1) and activin B, to a limited extent, inhibited the conversion of 17 alpha-OHP to the maturation-inducing hormone, 17 alpha,20 beta dihydroxy-4-pregnen-3-one. In conclusion, this study shows that TGF is expressed in the goldfish ovary, and that both activin and TGFbeta affect steroid production, which provides evidence that these members of the TGFbeta superfamily may act as local regulators of ovarian function in a teleost. PMID- 12765655 TI - Ecdysteroid concentrations through various life-stages of the meiobenthic harpacticoid copepod, Amphiascus tenuiremis and the benthic estuarine amphipod, Leptocheirus plumulosus. AB - Endocrine function in arthropods has principally been characterized in insects and malacostracan crustaceans. However, meiofauna represent the most abundant metazoan marine taxa, with harpacticoid copepods comprising the second most abundant taxon. In addition, their diminutive biomass has made characterization of endocrine components difficult, so little is known about endocrine control of reproduction, molting, and growth in meiofauna. In this study, a sensitive fluorometric enzyme immunoassay (EIA) was utilized to quantify and compare the arthropod molting hormone, 20-hydroxyecdysone (20E), in various life-cycle and developmental stages of a laboratory reared meiobenthic copepod, Amphiascus tenuiremis, and in an amphipod, Leptocheirus plumulosus. In copepods, gravid females carrying late stage pre-hatch embryos contained significantly more 20E (390+/-252 fmol/female) than gravids carrying early (Stage-I) embryos (172+/-83 fmol/female). In contrast, ecdysteroid levels in Stage-I L. plumulosus gravid females (277+/-83 fmol/female) was greater than pre-hatch gravid females (146+/ 42). Stage-I embryos of both copepods (19+/-10) and amphipods (11+/-5 fmol/embryo) possessed lower ecdysteroid content than copepod (35+/-15) and amphipod (43+/-33 fmol/embryo) pre-hatch embryos. Ecdysteroid levels were also assessed in naupliar, juvenile, adult male and non-gravid female copepod life stages. In addition, ecdysteroids measured in field collected copepod species indicated gravid females possessed ecdysteroid levels similar to gravid A. tenuiremis. However, upon normalization of egg sac 20E content by brood size, embryos from larger broods contained lower levels of ecdysteroids when compared to embryos from smaller clutch sizes-indicating an inverse embryo/ecdysteroid relationship may exist across species. PMID- 12765656 TI - Interactions between ecology, demography, capture stress, and profiles of corticosterone and glucose in a free-living population of Australian freshwater crocodiles. AB - In this study we examined three aspects pertaining to adrenocortical responsiveness in free-ranging Australian freshwater crocodiles (Crocodylus johnstoni). First, we examined the ability of freshwater crocodiles to produce corticosterone in response to a typical capture-stress protocol. A second objective addressed the relationship between capture stress, plasma glucose and corticosterone. Next we examined if variation in basal and capture-stress-induced levels of plasma corticosterone was linked to ecological or demographic factors for individuals in this free-ranging population. Blood samples obtained on three field trips were taken from a cross-sectional sample of the population. Crocodiles were bled once during four time categories at 0, 0.5, 6, and 10h post capture. Plasma corticosterone increased significantly with time post-capture. Plasma glucose also significantly increased with duration of capture-stress and exhibited a positive and significant relationship with plasma corticosterone. Significant variation in basal or stress induced levels of corticosterone in crocodiles was not associated with any ecological or demographic factors including sex, age class or the year of capture that the crocodiles were sampled from. However, three immature males had basal levels of plasma corticosterone greater than 2 standard deviations above the mean. While crocodiles exhibited a pronounced adrenocortical and hyperglycaemic response to capture stress, limited variation in adrenocortical responsiveness due to ecological and demographic factors was not evident. This feature could arise in part because this population was sampled during a period of environmental benigness. PMID- 12765657 TI - Cloning and mRNA expression of guanylin, uroguanylin, and guanylyl cyclase C in the Spinifex hopping mouse, Notomys alexis. AB - Guanylin and uroguanylin are peptides that activate guanylyl cyclase C (GC-C) receptors in the intestine and kidney, which causes an increase in the excretion of salt and water. The Spinifex hopping mouse, Notomys alexis, is a desert rodent that can survive for extended periods without free access to water and it was hypothesised that to conserve water, the expression of guanylin, uroguanylin, and GC-C would be down-regulated to reduce the excretion of water in urine and faeces. Accordingly, this study examined the expression of guanylin, uroguanylin, and GC-C mRNA in Notomys under normal (access to water) and water-deprived conditions. Initially, guanylin and uroguanylin cDNAs encoding the full open reading frame were cloned and sequenced. A PCR analysis showed guanylin and uroguanylin mRNA expression in the small intestine, caecum, proximal and distal colon, heart, and kidney. In addition, a partial GC-C cDNA was obtained and GC-C mRNA expression was demonstrated in the proximal and distal colon, but not the kidney. Subsequently, a semi-quantitative PCR method showed that water deprivation in Notomys caused a significant increase in guanylin and uroguanylin mRNA expression in the distal colon, and in guanylin and GC-C mRNA expression in the proximal colon. No significant difference in guanylin and uroguanylin mRNA expression was observed in the kidney. The results of this study indicate that there is, in fact, an up-regulation of the colonic guanylin system in Notomys after 7 days of water deprivation. PMID- 12765659 TI - Hospital malnutrition: is a disease or lack of food? PMID- 12765660 TI - Immunonutrition in the intensive care unit. A systematic review and consensus statement. AB - OBJECTIVE: To systematically review the effects of enteral nutrition with pharmaconutrients-enriched diets in critically ill patients and to establish recommendations for their use. DATA SOURCES: Computerized bibliographic search of published research and citation review of relevant articles. STUDY SELECTION: Randomized clinical trials of critically ill patients treated with enteral nutrition comparing diets enriched with pharmaconutrients vs not enriched diets were included. Infectious complications and outcome variables (days on mechanical ventilation, ICU and hospital length of stay and mortality) were evaluated. Studies were classified in four subgroups according to the patient's primary diagnosis: surgical, trauma, burned or medical. DATA EXTRACTION: A group of experts in methodology performed data extraction and statistical processes. A global analysis of the studies was done and also a separate study for each subgroup. Results of the meta-analysis were discussed within a 'clinical group' of clinicians with experience in the nutritional support of ICU patients, in order to find agreement about recommendations for the use of pharmaconutrients enriched diets in critically ill patients. RESULTS: Independent review of 267 articles identified 26 relevant primary studies. Global results indicate that there was a reduction in infection rate in the pharmaconutrition group, considering the appreciated lower incidence in abdominal abscesses (OR: 0.26, CI: 0.12-0.55) (P=0.005), nosocomial pneumonia (OR: 0.54, CI: 0.35-0.84) (P=0.007) and bacteremia (OR: 0.45, CI: 0.35-0.84) (P=0.0002). Also, patients treated with pharmaconutrition diets have a reduction in time on mechanical ventilation (mean 2.25 days, CI: 0.5-3.9) (P=0.009), ICU length of stay (mean reduction of 1.6 days, CI: 1.9-1.2) (P<0.0001) and hospital length of stay (mean reduction of 3.4 days, CI: 4.0-2.7) (P<0.0001). No effects were appreciated on mortality (OR: 1.10, CI: 0.85-1.42) (P=0.5). Nevertheless, the separate analysis for each subgroup showed that the reported beneficial effects were not the same for each patient population. Also, the clinician panel of experts identifies several problems in the published data about enteral pharmaconutrition in critically ill patients. In spite of the subgroup differences and of the problems detected, the clinician group considered that the appreciated results could support a Grade B recommendation for the use of these formulas in ICU patients. CONCLUSIONS: Considering the beneficial effects and the absence of detrimental ones, the use of diets enriched with pharmaconutrients could be recommended in ICU patients requiring enteral feeding. Nevertheless, more investigation is needed in this field in order to find the more appropriate population of patients that can benefit from this nutritional therapy. PMID- 12765661 TI - The impact of malnutrition on morbidity, mortality, length of hospital stay and costs evaluated through a multivariate model analysis. AB - Malnutrition has been identified as affecting patient outcome. The purpose of this study was to correlate the nutritional status of hospitalized patients with their morbidity, mortality, length of hospital stay and costs. The patients were nutritionally assessed within the first 72 h of hospital admission. The patients' charts were surveyed on the incidence of complications and mortality. Hospital costs were calculated based on economic tables used by insurance companies. Multivariate logistic regression analysis and the Cox regression model were used to identify possible confounding factors. A P<0.05 was considered statistically significant. The mean age was 50.6+/-17.3 years with 50.2% being male. The incidence of complications in the malnourished was 27.0% [Relative risk (RR)=1.60]. Mortality in the malnourished patients was 12.4% vs 4.7% in the well nourished (RR = 2.63). Malnourished patients stayed in the hospital for 16.7+/ 24.5 days vs 10.1+/-11.7 days in the nourished. Hospital costs in malnourished patients were increased up to 308.9%. It was concluded that malnutrition, as analyzed by a multivariate logistic regression model, is an independent risk factor impacting on higher complications and increased mortality, length of hospital stay and costs. PMID- 12765662 TI - Hospital food: a survey of patients' perceptions. AB - BACKGROUND & AIMS: Most hospitalised patients are dependent on hospital food for their nutritional requirements. We surveyed hospitalised patients to obtain their opinions of hospital food in order to improve menu planning and food delivery. METHODS: Three hundred and seventeen questionnaires were distributed to patients on an oral diet in two Swiss hospitals. Questions assessed eating habits, appetite, satisfaction with menus, food preferences and presentation, understanding of choices available and preferred choices. RESULTS: Three hundred and nine (97.5%) questionnaires were completed and analysed. Two hundred and sixty-five (86%) respondents were satisfied or very satisfied with hospital food. Two hundred and forty-one (78%) were satisfied with the way in which the food was served. There was a negative correlation (P=0.005) between duration of hospital stay and satisfaction with the food provided: 121 (39%) respondents stated that their appetite was the same as at home, and 50% stated that it had decreased during their time in the hospital. Eighty-seven (28%) patients said they ate all the food served, 148 (48%) ate most of it, and 68 (22%) ate only a small proportion. Patients felt that the temperature, appearance and aroma of the food were particularly important. CONCLUSION: Based on the findings of this survey we have made recommendations for improvements in hospital food and its presentation. PMID- 12765663 TI - Relationship between metabolism and peripheral blood mononuclear cell mitochondrial complex I activity before and after a short-term refeeding in weight-losing cancer patients. AB - BACKGROUND AND AIMS: Previous investigations showed that mitochondrial complex I activity seems to be a specific marker of dietary malnutrition in human. Since cancer has a more complex etiology than simple calorie deficiency, the aim of this study was to investigate the relationship between mitochondrial complex I activity and cancer. METHODS: Nine cancer patients (CaPs) with weight loss and 14 age-matched healthy volunteers (HVs) were recruited. Body mass index (BMI), body composition as well as resting energy expenditure (REE) and RQ were measured. Mitochondrial complex I activity was measured as described previously in isolated peripheral blood mononuclear cells. Six patients were investigated again after 7 days of refeeding. RESULTS: Weight loss in CaPs was mainly due to a loss of fat mass (FM), while fat-free mass (FFM) was preserved. The RQ was significantly lower in CaPs compared to HVs (P<0.001) and peripheral blood mononuclear cell complex I activity was significantly correlated with the %FM and RQ in CaPs. Furthermore, complex I activity increased significantly after 1 week of refeeding. CONCLUSIONS: Our study showed that mitochondrial complex I activity was inversely correlated to parameters of increased fat oxidation and reduced FM, which are indices of dietary insufficiency rather than loss of lean body mass, which is an index of increased catabolism in cancer. PMID- 12765664 TI - The effect of an oral glucose load on sodium and water excretion after rapid intravenous infusion of 0.9% (w/v) saline. AB - BACKGROUND & AIMS: Previous studies have suggested that oral or intravenous glucose enhances salt and water retention following a saline load. To test this, we studied the effects of an oral glucose load on urinary sodium and water excretion and serum biochemistry in response to a 2l intravenous infusion of 0.9% saline in normal subjects. METHODS: A crossover study was conducted on six male volunteers. On one occasion, they received 2l 0.9% saline intravenously over 1h. A week later, they were given 100ml 50% dextrose orally prior to the same infusion. Subjects passed urine before start of the infusion. Body weight, haematocrit and serum biochemistry were recorded preinfusion and hourly for 6h. Urine was collected for 6h postinfusion and analysed for sodium, potassium and osmolality. RESULTS: The six subjects had a mean (SE) age of 20.9 (0.4) years and BMI of 22.7 (0.2). Median (IQR) water balance over 6h was 1462 (1005-1650)ml after saline and 1203 (989-1735)ml after glucose and saline (NS). Urinary sodium and potassium excretion on the two occasions over 6h were 76 (69-111) vs 74 (92 174)mmol and 31 (29-40) vs 30 (20-36)mmol, respectively (NS). Using repeated measures testing, there was no significant difference in body weight, haematocrit, serum albumin, sodium, potassium, chloride, osmolality and blood glucose measured at hourly intervals on the two occasions. CONCLUSIONS: In contrast to previous literature, in normal subjects, an additional oral glucose load does not appear to have an effect on urinary sodium excretion or serum biochemistry after a rapid 2l infusion of 0.9% saline. This does not preclude an effect under conditions of prior starvation or injury. PMID- 12765665 TI - Home enteral nutrition in adults: a European multicentre survey. AB - AIMS: This study was undertaken to report indications and practice of home enteral nutrition (HEN) in Europe. METHODS: A questionnaire on HEN practice was sent to 23 centres from Belgium (B), Denmark (D), France (F), Germany (G), Italy (I), Poland (P), Spain (S) and the United Kingdom (UK). This involved adult patients newly registered in HEN programme from 1 January 1998 to 31 December 1998. RESULTS: A total of 1397 patients (532 women, 865 men) were registered. The median incidence of HEN was 163 patients/million inhabitants/year (range: 62 457). Age distribution was 7.5%, 16-40 years; 37.1%, 41-65 years; 34.5%, 66-80 years and 20.9% >80 years. The chief underlying diseases were a neurological disorder (49.1%), or head and neck cancer (26.5%); the main reason for HEN was dysphagia (84.6%). A percutaneous endoscopic gastrostomy (58.2%) or a naso gastric tube (29.3%) were used to infuse commercial standard or high energy diets (65.3%), or fibre diets (24.5%); infusion was cyclical (61.5%) or bolus (34.1%). Indications and feeds were quite similar throughout the different centres but some differences exist concerning the underlying disease. There was greater variation in the choice of tubes and mode of infusion. In F, G, I, S, and UK, costs of HEN are fully funded. In B, D, and P patients have to pay part or all of the charges. CONCLUSIONS: In Europe, HEN was utilised mainly in dysphagic patients with neurological disorders or cancer, using a standard feed via a PEG. However, there were important differences among the countries in the underlying diseases treated, the routes used, the mode of administration and the funding. PMID- 12765666 TI - Bedside placement of nasojejunal tubes: a randomised-controlled trial of spiral- vs straight-ended tubes. AB - BACKGROUND AND AIMS: The success rate of unguided nasojejunal feeding tube insertion is low, thus often requiring endoscopic or radiological assistance. The spiral end of the Bengmark nasojejunal tube is supposed to aid post-pyloric placement, but no comparative trial has been performed. METHODS: Patients requiring nasojejunal feeding were randomised to have either Medicina (straight) or Bengmark (spiral) nasojejunal tube placed after stratification into those with normal gastric emptying or clinical evidence of delayed gastric emptying. Nasojejunal tubes were placed at the bedside in a standard fashion without radiological guidance by the same person for pre- and/or post-operative feeding. Bolus intravenous metaclopromide (10mg) was given prior to insertion in the abnormal gastric emptying group. Abdominal radiographs were obtained at 4 and 24h, and the primary end-point was jejunal placement at 24h. RESULTS: Forty-seven patients were randomised of which 17 (11 straight, 6 spiral) could not tolerate the nasojejunal tube. Of the 30 remaining patients, 16 had normal gastric emptying. In patients with normal gastric emptying, successful placement at 24h was achieved in 78% (spiral tube), vs 14% (straight tube) (P=0.041). In the abnormal gastric emptying group, success rates were 57% and 0%, respectively (P=0.07). CONCLUSION: Spiral nasojejunal tubes are preferable to straight tubes for bedside unguided post-pyloric feeding in patients with normal gastric emptying. PMID- 12765667 TI - Anaemia in the first but not in the second or third trimester is a risk factor for low birth weight. AB - OBJECTIVE: To assess pregnancy outcome in women with anaemia during pregnancy. METHODS: The study design involved a retrospective chart review of all women registering for prenatal care in the area of Kuopio University Hospital between 1990 and 2000. A haemoglobin concentration below 100g/l was used as a cutoff for anaemia and affected women (N=597) were stratified by the trimester at which anaemia was diagnosed. Multiple regression analysis was used to compare obstetric outcomes in the study groups and in non-anaemic women (N=22,202). RESULTS: The frequency of anaemia was 2.6%, with 0.3% occurring in the first trimester. After controlling for confounding factors, anaemia detected in the first trimester was associated with low-birth-weight infants (OR=3.14, 95% CI: 1.35-7.28) whereas the mid- and third-trimester anaemia groups showed no significantly different outcomes when compared with the non-anaemic women. First trimester anaemia was not significantly associated with small birth weight for gestational age (OR=0.98, 95% CI: 0.41-2.17) or with premature delivery <37 weeks (OR=1.80, 95% CI: 0.72-4.49). CONCLUSIONS: Maternal anaemia detected in the first trimester is associated with low birth weight. PMID- 12765668 TI - The effect of immunonutrition on bacterial translocation, and intestinal villus atrophy in experimental obstructive jaundice. AB - BACKGROUND & AIMS: Spontaneous bacterial infection and septicemia due to increased bacterial translocation (BT) in patients with obstructive jaundice result in significant morbidity and mortality. The present study evaluates the effects of enteral nutrition with immune enhancing feeds on BT and intestinal villus histopathology promoted by obstructive jaundice. METHODS: Fifty male Wistar-albino rats weighing 250-300g were assigned into five equal groups of 10. Animals in Groups I, II, and III were fed with standard chow, those in Group IV were given glutamine 1g/kg/day and the remaining 10 animals in Group V were fed with an arginine, omega-3 fatty acids, and RNA-supplemented enteral diet for (1g/kg/day amino acid and 230 kcal/kg) 7 days preoperatively. Group I underwent sham operation and the remaining animals in all other groups underwent common bile duct ligation. After operation, Group I had standard chow, Groups II and IV had glutamine, Groups III and V had an arginine omega-3 fatty acids, and RNA supplemented enteral diet for 7 days. All animals were sacrificed on the 8th postoperative day and evaluated both biochemically and histopathologically. Samples from blood, liver, mesenteric lymph nodes and spleen were cultured under aerobic conditions. RESULTS: Significantly less BT was observed in groups fed with an arginine, omega-3 fatty acids, and RNA-supplemented enteral diet or glutamine in pre-and postoperative periods as compared to others (P<0.001). Histologic evaluation also showed significant reduction in villus atrophy in these groups. CONCLUSIONS: Enteral immunonutrition using glutamine or arginine, omega-3 fatty acids, and RNA-supplemented enteral diet during both pre-and postoperative periods seems to reduce BT and decrease atrophy of intestinal mucosal villi in rats with obstructive jaundice. PMID- 12765669 TI - Nutritional support in acute stroke: the impact of evidence-based guidelines. AB - BACKGROUND AND AIMS: Stroke patients experience multiple impairments which impair ability to eat and render them vulnerable to the deleterious sequelae of malnutrition. This study aimed to develop, implement and evaluate evidence-based guidelines for nutrition support following acute stroke using a multifaceted change management strategy. METHODS: Prospective quasi-experimental design. Documentation of two groups of 200 acute stroke patients admitted to medical and care of the elderly wards of an acute NHS Trust in South London was surveyed using a checklist before and after implementation of 24 guidelines for nutritional screening, assessment and support. Guidelines were based on systematic literature review and developed by consensus in a nurse-led multiprofessional group; implemented via a context-specific, multifaceted strategy including opinion leaders and educational programmes linked to audit and feedback. STAFF OUTCOMES: Compliance with guidelines by doctors, nurses, therapists. PATIENT OUTCOMES: Changes in Barthel Index scores and Body Mass Index in hospital, infective complications, length of stay, discharge destination. RESULTS: Statistically significant improvements in compliance with 15 guidelines occurred in the post-test group. Infective episodes showed a significant reduction in the post-test group but other patient outcomes were unaffected. CONCLUSIONS: Implementation of evidence-based guidelines for nutritional support following acute stroke using a multifaceted strategy was associated with improvements in documented practice and selected patient outcomes. PMID- 12765670 TI - Comparison of a high-protein disease-specific enteral formula with a high-protein enteral formula in hyperglycemic critically ill patients. AB - AIMS: To determine whether a specific high-protein enteral formula with a similar caloric percentage of fat and carbohydrates achieves greater control over glycemic levels and reduces insulin requirements in hyperglycemic critically ill patients when compared to a control high-protein enteral formula. DESIGN: A prospective, randomized, controlled, single-blind trial in two University Hospital Intensive Care Units in Spain. METHODS: We enrolled 50 patients with diabetes mellitus or stress hyperglycemia with basal glycemia > or =160 mg/dl and indication for enteral nutrition > or =5 days. Patients with severe kidney failure, liver failure or obesity were excluded from the study. In the first 48 h of admission, after randomization, 26 patients received the study diet and 24 patients received the control diet. The variables were monitored for 14 days. The Harris-Benedict formula with a fixed stress factor of 1.2 was used to calculate caloric needs. Insulin was administered by continuous infusion. An intention-to treat analysis was performed. RESULTS: On admission, there were no differences between the study and control group in plasma glucose levels (mg/dl) (190.9+/-45 vs 210.3+/-63) and capillary glucose levels (mg/dl) (226.1+/-73 vs 213.8+/-67). After the feeding trial, there were differences between the study and control group in plasma glucose levels (mg/dl) (176.8+/-44 vs 222.8+/-47, P=0.001), capillary glucose levels (mg/dl) (163.1+/-45 vs 216.4+/-56, P=0.001), insulin requirements/day (IU) 8.73 (2.3-27.5) vs 30.2 (21.5-57.1) (P=0.001), insulin/received carbohydrates (UI/g) 0.07 (0.02-0.22) vs 0.18 (0.11-0.35) (P=0.02) and insulin/received carbohydrates/kg 0.98 (0.26-3.59) vs 2.13 (1.44 4.58) (P=0.04). These differences remain in a day-to-day comparison. There were no differences in the analytical tests, or in digestive or infectious complications. Intensive Care Unit length of stay, mechanical ventilation and mortality were similar in both groups. CONCLUSIONS: Hyperglycemic critically ill patients fed with a high-protein diet with a similar caloric percentage of fat and carbohydrates show a significant reduction in plasma glucose levels, capillary glucose levels and insulin requirements in comparison to patients on a conventional high-protein diet. This better glycemic control do not modify Intensive Care Unit length of stay, infectious complications, mechanical ventilation and mortality. PMID- 12765671 TI - A comparison of mid upper arm circumference, body mass index and weight loss as indices of undernutrition in acutely hospitalized patients. AB - A nutritional supplementation trial (Vlaming et al., Clin Nutr 2001; 20: 517) enabled us to assess the nutrition of 1561 patients on emergency admission to hospital. Patients acutely admitted to the 15 relevant medical, surgical and orthopaedic wards were identified. Mid upper arm circumference (MUAC) measurements were obtained in 95% (848 m, 635f) patients. For clinical reasons, Body mass index (BMI) was assessable in only 44% patients (408 m, 285f). Data on three month weight loss were obtainable in 509 patients. These measurements combined to demonstrate that 18.3% of patients were undernourished (At least one of : BMI<20 kg/m(2) or MUAC<25 cm or loss of weight > or =10%). There was a close relationship between BMI and MUAC. Regression equations (excluding age)were for men : BMI=1.01 x MUAC-4.7, (R(2)=0.76), and for women BMI=1.10 x MUAC-6.7, (R(2)=0.76). After adjustment for age, weight loss > or =10% was the most significant of the three as a predictor of mortality. Among patients in whom weight loss was not recorded MUAC was a significant predictor of mortality either alone (P=0.002) or after adjustment for BMI (P=0.007), but BMI was not significant. All three measures, even when adjusted for age and sex, were poor predictors of hospital stay although MUAC was significant in the larger group with a MUAC measure (R(2)=0.7% P<0.001). MUAC correlates closely with BMI, is easier to measure and predicts poor outcome better. PMID- 12765672 TI - Anti-oxidant properties of N-acetyl-L-cysteine do not improve the immune resistance of mice fed dietary lipids to Listeria monocytogenes infection. AB - BACKGROUND & AIMS: Current knowledge of the potential effects that several dietary lipids exert on immune functions indicates that these substances participate actively in the modulation of immune system by which they contribute to the improvement of the conditions of patients suffering from inflammatory disorders. However, long-chain n-3 polyunsaturated fatty acids induce an immunosuppressive status that leads to a reduction of the host natural resistance to infectious agents as well as to an enhancement of oxidative damage. Hence, the present study has been designed to evaluate the effects on the immune system of the antioxidant N-acetyl-L-cysteine (NAC) in mice fed dietary lipids and infected with Listeria monocytogenes. METHODS: Balb/c mice were fed for 4 weeks with diets containing either olive oil (OO, 20% by weight), fish oil (FO, 20% by weight) or hydrogenated coconut oil (HCO, 20% by weight). After dietary lipid administration mice were experimentally infected with L. monocytogenes or treated with NAC (25mg/ml intraperitoneally). RESULTS: NAC at a concentration of 1mM promoted a loss of cell viability, although no differences were observed among the four groups. After injection of NAC in combination with L. monocytogenes, 25% of mice fed a low-fat (LF) diet survived. However, in the groups fed dietary lipids no effect on survival of mice was found. NAC participated in the reduction of superoxide anion generation measured with nitroblue tetrazolium (NBT) in the group fed a FO diet. Finally, NAC reduced the recovery of L. monocytogenes from spleen of mice fed diets containing LF or HCO. CONCLUSIONS: On the basis of these results, we can confirm that the administration of NAC improves survival in mice fed LF diet, whereas a reduction in the generation of superoxide radicals was measured in mice fed a FO diet and infected with L. monocytogenes. Similarly, bacterial recovery was diminished in mice fed diets containing LF or HCO. Hence, these data reveal a beneficial effect of NAC in mice fed LF or HCO and a detrimental action of this antioxidant in mice fed diets containing FO or OO. PMID- 12765673 TI - Nutritional risk screening (NRS 2002): a new method based on an analysis of controlled clinical trials. AB - BACKGROUND & AIMS: A system for screening of nutritional risk is described. It is based on the concept that nutritional support is indicated in patients who are severely ill with increased nutritional requirements, or who are severely undernourished, or who have certain degrees of severity of disease in combination with certain degrees of undernutrition. Degrees of severity of disease and undernutrition were defined as absent, mild, moderate or severe from data sets in a selected number of randomized controlled trials (RCTs) and converted to a numeric score. After completion, the screening system was validated against all published RCTs known to us of nutritional support vs spontaneous intake to investigate whether the screening system could distinguish between trials with a positive outcome and trials with no effect on outcome. METHODS: The total number of randomized trials identified was 128. In each trial, the group of patients was classified with respect to nutritional status and severity of disease, and it was determined whether the effect of nutritional intervention on clinical outcome was positive or absent. RESULTS: Among 75 studies of patients classified as being nutritionally at-risk, 43 showed a positive effect of nutritional support on clinical outcome. Among 53 studies of patients not considered to be nutritionally at-risk, 14 showed a positive effect (P=0.0006). This corresponded to a likelihood ratio (true positive/false positive) of 1.7 (95% CI: 2.3-1.2). For 71 studies of parenteral nutrition, the likelihood ratio was 1.4 (1.9-1.0), and for 56 studies of enteral or oral nutrition the likelihood ratio was 2.9 (5.9-1.4). CONCLUSION: The screening system appears to be able to distinguish between trials with a positive effect vs no effect, and it can therefore probably also identify patients who are likely to benefit from nutritional support. PMID- 12765675 TI - From TRPs to SOCs, CCEs, and CRACs: consensus and controversies. PMID- 12765676 TI - The pore of TRP channels: trivial or neglected? AB - Ultimate proof that a protein contributes to the pore of an ion channel is demonstrating that pore properties can be altered by mutations to the putative pore-forming region. To date this has only been achieved for a few TRP proteins, and only within the TRPV subfamily. The location and structure of the pore region and selectivity filter of most TRP proteins, including all members of the TRPM and TRPC subfamilies, is currently unknown. In this review we give a short overview of the limited current knowledge about TRP channel pores and argue that further study is needed, not only for a better understanding of the mechanisms underlying cation permeation, but also to establish that all members of the TRP superfamily indeed function as bona fide ion channels. PMID- 12765677 TI - Proteins modulating TRP channel function. AB - TRP channels are involved in different signaling cascades; TRP channels can be activated via hormones and neurotransmitter in a receptor/G-protein-mediated manner or by osmotic, thermic or mechanic stimuli. The overall functional role of TRP channels within these processes of hormonal cellular control, nociception or cellular calcium homeostasis is still unclear, as these complex processes often involve macromolecular structures. Whereas the integration of Drosophila TRP in the phototransduction process is becoming clear, the understanding of the participation of mammalian TRP channels in signal transduction complexes is only beginning. TRP channels have been demonstrated to interact with PDZ domain proteins, and both scaffold and regulatory function have been shown for INAD, the PDZ domain protein of the Drosophila phototransduction complex. In mammalian cells, the interaction of NHERF and TRPC4 has been shown and it is anticipated that NHERF may abolish the apparent store-dependent regulation of TRPC4 and TRPC5. Whereas TRP channels and PDZ domain proteins form permanent heterodimeric proteins, the interaction of calcium-binding proteins is dependent on the calcium concentration and is, therefore, dynamic. The prototype of calcium-binding protein used for experiments is calmodulin; whether or not calmodulin is also the natural interaction partner of TRP channels is an open question. PMID- 12765678 TI - CRAC channels: activation, permeation, and the search for a molecular identity. AB - The Ca2+ release-activated Ca2+ (CRAC) channel is a highly Ca2+-selective store operated channel that is expressed in T lymphocytes, mast cells, and other hematopoietic cells. In T cells, CRAC channels are essential for generating the prolonged intracellular Ca2+ ([Ca2+](i)) elevation required for the expression of T-cell activation genes. Here we review recent work addressing CRAC channel regulation, pore properties, and the search for CRAC channel genes. Of the current models for CRAC current (I(CRAC)) activation, several new studies argue against a conformational coupling mechanism in which IP(3) receptors communicate store depletion to CRAC channels through direct physical interaction. The study of CRAC channels has been complicated by the fact that they lose activity in the absence of extracellular Ca2+. Attempts to maintain current size by removing intracellular Mg2+ have been found to unmask Mg2+-inhibited cation (MIC/MagNuM/TRPM7) channels, which have been mistaken in several studies for the CRAC channel. Recent studies under conditions that prevent MIC activation reveal that CRAC channels use high-affinity binding of Ca2+ in the pore to achieve high Ca2+ selectivity but have a surprisingly low conductance for both Ca2+ (approximately 10fS) and Na+ (approximately 0.2pS). Pore properties provide a unique fingerprint that provides a stringent test for potential CRAC channel genes and suggest models for the ion selectivity mechanism. PMID- 12765679 TI - On the endothelial cell I(SOC). AB - Ca2+ store depletion activates both Ca2+ selective and non-selective currents in endothelial cells. Recently, considerable progress has been made in understanding the molecular make-up and regulation of an endothelial cell thapsigargin activated Ca2+ selective current, I(SOC). Indeed, I(SOC) is a relatively small inward Ca2+ current that exhibits an approximate +40mV reversal potential and is strongly inwardly rectifying. This current is sensitive to organization of the actin-based cytoskeleton. Transient receptor potential (TRP) proteins 1 and 4 (TRPC1 and TRPC4, respectively) each contribute to the molecular basis of I(SOC), although it is TRPC4 that appears to be tethered to the cytoskeleton through a dynamic interaction with protein 4.1. Activation of I(SOC) requires association between protein 4.1 and the actin-based cytoskeleton (mediated through spectrin), suggesting protein 4.1 mediates the physical communication between Ca2+ store depletion and channel activation. Thus, at present findings indicate a TRPC4 protein 4.1 physical linkage regulates I(SOC) activation following Ca2+ store depletion. PMID- 12765680 TI - Localization and regulation of Ca2+ entry and exit pathways in exocrine gland cells. AB - Studies of Ca2+ transport pathways in exocrine gland cells have been useful, chiefly because of the polarized nature of the secretory epithelial cells. In pancreatic acinar cells, for example, Ca2+ reloading of empty intracellular stores can occur solely via Ca2+ entry through the basal part of the plasma membrane. On the other hand, the principal site for intracellular Ca2+ release with the highest concentration of inositol 1,4,5-trisphosphate (IP(3)) receptors is in the apical secretory pole close to the apical plasma membrane. This apical part of the plasma membrane contains the highest density of Ca2+ pumps and is therefore the principal site for Ca2+ extrusion. On the basis of the known properties of Ca2+ entry and exit pathways in exocrine gland cells, the mechanisms controlling Ca2+ exit and entry are discussed in relation to recent direct information about Ca2+ transport into and out of the endoplasmic reticulum (ER) and the mitochondria in these cells. PMID- 12765681 TI - Store-operated Ca2+-permeable non-selective cation channels in smooth muscle cells. AB - Over twenty years ago it was shown that depletion of the intracellular Ca2+ store in smooth muscle triggered a Ca2+ influx mechanism. The purpose of this review it to describe recent electrophysiological data which indicate that Ca2+ influx occurs through discrete ion channels in the plasmalemma of smooth muscle cells. The effect of external Ca2+ on the amplitude and reversal potential of whole-cell and single channel currents suggests that there are at least two, and probably more, distinct store-operated channels (SOCs) which have markedly different permeabilities to Ca2+ ions. Two activation mechanisms have been identified which involve Ca2+ influx factor and protein kinase C (PKC) activation via diacylglycerol. In addition, in rabbit portal vein cells there is evidence that stimulation of alpha-adrenoceptors can stimulate SOC opening via PKC in a store independent manner. There is at present little knowledge on the molecular identity of SOCs but it has been proposed that TRPC1 may be a component of the functional channel. We also summarise the data showing that SOCs may be involved in contraction and cell proliferation of smooth muscle. Finally, we highlight the similarities and differences of SOCs and receptor-operated cation channels that are present in native rabbit portal vein myocytes. PMID- 12765682 TI - Store-operated Ca2+ channels in prostate cancer epithelial cells: function, regulation, and role in carcinogenesis. AB - Ca2+ homeostasis mechanisms, in which the Ca2+ entry pathways play a key role, are critically involved in both normal function and cancerous transformation of prostate epithelial cells. Here, using the lymph node carcinoma of the prostate (LNCaP) cell line as a major experimental model, we characterize prostate specific store-operated Ca2+ channels (SOCs)--a primary Ca2+ entry pathway for non-excitable cells--for the first time. We show that prostate-specific SOCs share major store-dependent, kinetic, permeation, inwardly rectifying, and pharmacological (including dual, potentiation/inhibition concentration-dependent sensitivity to 2-APB) properties with "classical" Ca2+ release-activated Ca2+ channels (CRAC), but have a higher single channel conductance (3.2 and 12pS in Ca2+- and Na+-permeable modes, respectively). They are subject to feedback inhibition via Ca2+-dependent PKC, CaMK-II and CaM regulatory pathways and are functionally dependent on caveolae integrity. Caveolae also provide a scaffold for spatial co-localization of SOCs with volume-regulated anion channels (VRAC) and their Ca2+-mediated interaction. The TRPC1 and TRPV6 members of the transient receptor potential (TRP) channel family are the most likely molecular candidates for the formation of prostate-specific endogenous SOCs. Differentiation of LNCaP cells to an androgen-insensitive, apoptotic-resistant neuroendocrine phenotype downregulates SOC current. We conclude that prostate-specific SOCs are important determinants in the transition to androgen-independent prostate cancer. PMID- 12765683 TI - Retrograde activation of store-operated calcium channel. AB - Store-operated Ca2+ entry represents an important mechanism for refilling of a depleted intracellular-reticulum Ca2+ store following sustained activation of the IP3 receptor or ryanodine receptor RyR/Ca2+ release channel in the endoplasmic/sarcoplasmic reticulum (ER/SR). Recent studies have demonstrated the existence of store-operated Ca2+ channel (SOC) in muscle cells, whose activation process appears to be coupled to conformational changes of the RyR. Regulation of the plasma membrane (PM)-resided SOC by the SR-located RyR requires an integrity of the junctional membrane structure between SR and PM. Proteins that interact with RyR or influence the Ca2+ buffering capacity in the ER or SR lumen also participate in the activation process of SOC. Calsequestrin (CSQ) and calreticulin (CRT) are SR/ER-resident proteins, with highly negative charged regions at the carboxyl-terminal end that exhibit high buffering capacity for luminal Ca2+. CSQ and CRT not only modulate the intracellular Ca2+ release process but also might provide retrograde signals to regulate the function of SOC. The functional interplay between CSQ, RyR and SOC may serve essential roles of Ca2+ signaling in muscle contraction and development. A tight link between the expression of CRT and operation of SOC exist in certain cancer cells, where the reduced sensitivity to apoptosis may correlate with the altered function of SOC. PMID- 12765684 TI - TRP channels in Drosophila photoreceptors: the lipid connection. AB - The light-sensitive current in Drosophila photoreceptors is mediated by transient receptor potential (TRP) channels, at least two members of which (TRP and TRPL) are activated downstream of phospholipase C (PLC) in response to light. Recent evidence is reviewed suggesting that Drosophila TRP channels are activated by one or more lipid products of PLC activity: namely diacylglycerol (DAG), its metabolites (polyunsaturated fatty acids) or the reduction in phosphatidylinositol 4,5-bisphosphate (PIP(2)). The most compelling evidence for this view comes from analysis of rdgA mutants which are unable to effectively metabolise DAG due to a defect in DAG kinase. The rdgA mutation leads to constitutive activation of both TRP and TRPL channels and dramatically increases sensitivity to light in hypomorphic mutations of PLC and G protein. PMID- 12765685 TI - TRP gating is linked to the metabolic state and maintenance of the Drosophila photoreceptor cells. AB - The Drosophila light-activated channel TRP is the founding member of a large and diverse family of channel proteins that is conserved throughout evolution. In spite of much progress, the gating mechanism of TRP channels is still unknown. However, recent studies have shown multi-faceted functions of the Drosophila light-sensitive TRP channel that may shed light on TRP gating. Accordingly, metabolic stress, which leads to depletion of cellular ATP, reversibly activates the Drosophila TRP and TRPL channels in the dark in a constitutive manner. In several Drosophila mutants, constitutive activity of TRP channels lead to a rapid retinal degeneration in the dark, while genetic elimination of TRP protects the cells from degeneration. Additional studies have shown that TRPL translocates in a light-dependent manner between the signaling membranes and the cell body. This light-activated translocation is accompanied by reversible morphological changes leading to partial and reversible collapse of the microvillar signaling membranes into the cytosol, which allows turnover of signaling molecules. These morphological changes are also blocked by genetic elimination of TRP channels. The link of TRP gating to the metabolic state and maintenance of cells makes cells expressing TRP extremely vulnerable to metabolic stress via a mechanism that may underlie retinal degeneration and neuronal cell death upon malfunction. PMID- 12765686 TI - The venerable inveterate invertebrate TRP channels. AB - The transient receptor potential (TRP) superfamily is subdivided into four main classes of cation channels, TRPC, TRPV, TRPM and TRPN, each of which includes members in worms, flies, mice and humans. While the biophysical features of many of the mammalian channels have been described, relatively little is known concerning the biological roles of these channels. Forward genetic screens in Drosophila melanogaster and Caenorhabditis elegans have led to the identification of the founding members of each of these four subfamilies. Moreover, phenotypic analyses of invertebrate mutants have contributed greatly to our understanding of the roles of TRP proteins. A recurring theme is that many of these proteins function in sensory signaling processes ranging from vision to olfaction, osmosensation, light touch, social feeding, and temperature- and mechanically induced nociception. In addition, at least one invertebrate TRP protein is required for cell division. As many of these functions may be conserved among the mammalian TRPs, the invertebrate TRPs offer valuable genetic handles for characterizing the functions of these cation channels in vivo. PMID- 12765687 TI - A comparison of the genes coding for canonical TRP channels and their M, V and P relatives. AB - The mammalian transient receptor potential (TRP) protein gene family consists of a diverse group of cation channels that currently contain at least 26 members. The physiologic functions of many remain unknown. They are structurally similar to Drosophila TRP and have a wide tissue distribution. In the present report, we compare the chromosomal locations, the gene, and primary structures of each of these 26 human TRP family members. Based on primary amino acid analyses, these channels comprise four different subfamilies: C- (canonical or classical), V- (or vanilloid receptor related), M- (melastatin related), and P (PKD)-type. The highest homology within each subfamily and between subfamilies exists in the predicted ion channel domains. Belonging to a given subfamily, however, does not determine the activating stimuli. This is exemplified by the V- and M subfamilies, both of which have members that respond to temperature and osmolarity. TRP genes vary in their intron-exon organization, with the greatest diversity in the P subfamily. Chromosomal organization analyses revealed that two TRP members are found as direct repeats; TRPV3 follows TRPV1 and TRPV6 follows TRPV5. Both of these duplications appear to be recent as TRPV1 and V3 are more similar to each other than to other members of the TRPV subfamily. The same holds true for TRPV5 and V6. The article presents complication of comparisons including exon-intron boundaries, the amino acid sequence alignments, and the chromosomal organization of each of the presently known TRP channels. PMID- 12765688 TI - TRPC1 store-operated cationic channel subunit. AB - TRPC1 is a membrane protein that is highly conserved in mammals, amphibians and birds. It is widely expressed in cells throughout the body including in the heart and nervous system. Amino acid sequence analysis and over-expression studies indicate it is an ion channel that allows the transmembrane flux of small cations including sodium and calcium. In some cell types it is apparent that at least a fraction of TRPC1 exists in the plasma membrane. Inhibition of TRPC1 expression or block by TRPC1-specific antibody leads to attenuation of the plasma membrane calcium influx that occurs in response to depletion of calcium levels in sarcoplasmic or endoplasmic reticulum. TRPC1 would, therefore, seem to be a key subunit of store-operated channels (SOCs). TRPC1 is, nevertheless, unlikely to act alone. There is good evidence that it can heteromultimerise with the related proteins TRPC4, TRPC5 and polycystin-2; a tetrameric arrangement is envisaged, but not demonstrated. Like its relative in Drosophila, TRPC1 looks likely to function in a signalplex, a protein complex including inositol 1,4,5-triphosphate (IP(3)) receptor, plasma membrane calcium-ATPase, caveolin-1 and calmodulin. Its localisation in membranes is punctate and associated with functionally discrete calcium signals. TRPC1's function may not only be linked to SOCs but also to other cellular events including the nuclear translocation of the NFAT transcription factor. There is still much to be learned about this fundamental protein. PMID- 12765689 TI - TRPC4 and TRPC5: receptor-operated Ca2+-permeable nonselective cation channels. AB - The seven mammalian channels from the classical (TRPC) subfamily of transient receptor potential (TRP) channels are thought to be receptor-operated cation channels activated in a phospholipase C (PLC)-dependent manner. Based on sequence similarity, TRPC channels can be divided into four subgroups. Group 4 comprises TRPC4 and TRPC5, and is most closely related to group 1 (TRPC1). The functional properties observed following heterologous expression of TRPC4 or TRPC5 in mammalian cells are contradictory and, therefore, controversial. In our hands, and in several independent studies, both channels, probably as homotetramers, form receptor-operated, Ca2+-permeable, nonselective cation channels activated independently of inositol 1,4,5-trisphosphate (InsP(3)) receptor activation or Ca2+ store-depletion. As heteromultimers with TRPC1, TRPC4 and TRPC5 form receptor-operated, Ca2+-permeable, nonselective cation channels with biophysical properties distinct from homomeric TRPC4 or TRPC5. In other studies, TRPC4 and TRPC5 have been shown to be store-operated channels, with moderate to high Ca2+ permeabilities. At present there is no clear explanation for these major differences in functional properties. To date, little is known as to which native cation channels are formed by TRPC4 and TRPC5. Endothelial cells from TRPC4(-/-) mice lack a highly Ca2+-permeable, store-dependent current, and data support a role for TRPC4 in endothelium-mediated vasorelaxation. A similar current in adrenal cortical cells is reduced by TRPC4 antisense. From similarities in the properties of the currents and expression of appropriate isoforms in the tissues, it is likely that heteromultimers of TRPC1 and TRPC4 or TRPC5 form receptor operated nonselective cation channels in central neurones, and that TRPC4 contributes to nonselective cation channels in intestinal smooth muscle. PMID- 12765690 TI - The TRPC3/6/7 subfamily of cation channels. AB - The mammalian transient receptor potential (TRP) proteins consist of a superfamily of Ca2+-permeant non-selective cation channels with structural similarities to Drosophila TRP. The TRP superfamily can be divided into three major families, among them the "canonical TRP" family (TRPC). The seven protein products of the mammalian TRPC family of genes (designated TRPC1-7) share in common the activation through PLC-coupled receptors and have been proposed to encode components of native store-operated channels in different cell types. In addition, the three members of the TRPC3/6/7 subfamily of TRPC channels can be activated by diacylglycerol analogs, providing a possible mechanism of activation of these channels by PLC-coupled receptors. This review summarizes the current knowledge about the mechanism of activation of the TRPC3/6/7 subfamily, as well as the potential role of these proteins as components of native Ca2+-permeant channels. PMID- 12765691 TI - FRET-based analysis of TRPC subunit stoichiometry. AB - By analogy to other cation channel subunits with six transmembrane-spanning domains, the seven members of the "classical" or "canonical" transient receptor potential channels (TRPC) family are believed to assemble into homo- or heterotetrameric complexes. These complexes have been verified by classical methods such as coimmunoprecipitation, crosslinking analysis or functional assays applying dominant negative pore mutants. More recently, fluorescence resonance energy transfer (FRET)-a measure for the close proximity of fluorescent molecules has become instrumental in monitoring protein assembly in living cells. Here we demonstrate further possibilities and verification procedures of the FRET technology to test the assembly of ion channel subunits. Temporally and spatially resolved FRET imaging demonstrates an early assembly of TRPC subunits in the endoplasmic reticulum and the Golgi apparatus. Confocal FRET imaging verifies FRET signals over the plasma membrane at high spatial resolution. Taking advantage of the quantitative analysis of digital video imaging, we demonstrate that FRET between TRPC subunits is only poorly concentration-dependent. Moreover, a correlation between the efficiency of energy transfer and the molar ratio of the FRET donor to the acceptor was exploited to verify the tetrameric stoichiometry of TRPC complexes. Finally, we introduce a competition-FRET assay to test the ability of wild-type TRPC subunits to recruit fluorescent TRPC subunits into separate channel complexes. PMID- 12765692 TI - Vertebrate and invertebrate TRPV-like mechanoreceptors. AB - Our senses of touch, hearing, and balance are mediated by mechanosensitive ion channels. In vertebrates, little is known about the molecular composition of these mechanoreceptors, an example of which is the transduction channel of the inner ear's receptor cells, hair cells. Members of the TRP family of ion channels are considered candidates for the vertebrate hair cell's mechanosensitive transduction channel and here we review the evidence for this candidacy. We start by examining the results of genetic screens in invertebrates that identified members of the TRP gene family as core components of mechanoreceptors. In particular, we discuss the Caenorhabditis elegans OSM-9 channel, an invertebrate TRPV channel, and the Drosophila melanogaster TRP channel NOMPC. We then evaluate basic features of TRPV4, a vertebrate member of the TRPV subfamily, which is gated by a variety of physical and chemical stimuli including temperature, osmotic pressure, and ligands. Finally, we compare the characteristics of all discussed mechanoreceptive TRP channels with the biophysical characteristics of hair cell mechanotransduction, speculating about the possible make-up of the elusive inner ear mechanoreceptor. PMID- 12765693 TI - TRPV channels as temperature sensors. AB - The past year has seen a doubling in the number of heat-sensitive ion channels to six, and four of these channels are from the TRPV family. These channels characteristically have Q(10) values of >10 above the thermal threshold, very different from the Q(10) values of 1.5-2.0 seen in most ion channels. Cells expressing TRPV1 show similar temperature sensitivity to small capsaicin sensitive nociceptor neurons, consistent with these neurons expressing homomers of TRPV1. A-delta fibres exhibit properties that may be explained by TRPV2 containing channels which is present in large diameter sensory neurons that do not express TRPV1. TRPV3 has a lower temperature threshold and may contribute to warm-sensitive channels together with TRPV1. Warm sensation may also be transduced by TRPV4 expressing sensory neurons and hypothalamic neurons. We can now look forward to further work defining the properties of the recombinant channels in more detail and a re-analysis of endogenous i(heat) currents in thermosensitive neurons and other cells. Data from the study of mice in which TRPV2, TRPV3 or TRPV4 have been deleted are also eagerly awaited. PMID- 12765694 TI - Modulation of TRPV4 gating by intra- and extracellular Ca2+. AB - We have studied the modulation of gating properties of the Ca2+-permeable, cation channel TRPV4 transiently expressed in HEK293 cells. The phorbol ester 4alphaPDD transiently activated a current through TRPV4 in the presence of extracellular Ca2+. Increasing the concentration of extracellular Ca2+ ([Ca2+](e)) reduced the current amplitude and accelerated its decay. This decay was dramatically delayed in the absence of [Ca2+](e). It was also much slower in the presence of [Ca2+](e) in a mutant channel, obtained by a point mutation in the 6th transmembrane domain, F707A. Mutant channels, containing a single mutation in the C-terminus of TRPV4 (E797), were constitutively open. In conclusion, gating of the 4alphaPDD activated TRPV4 channel depends on both extra- and intracellular Ca2+, and is modulated by mutations of single amino acid residues in the 6th transmembrane domain and the C-terminus of the TRPV4 protein. PMID- 12765695 TI - The epithelial calcium channels, TRPV5 & TRPV6: from identification towards regulation. AB - The epithelial calcium channels, TRPV5 and TRPV6, have been extensively studied in epithelial tissues controlling the Ca(2+) homeostasis and exhibit a range of distinctive properties that distinguish them from other TRP channels. This review focuses on the tissue distribution, the functional properties, the architecture and the regulation of the expression and activity of the TRPV5 and TRPV6 channel. PMID- 12765696 TI - The TRPV6 gene, cDNA and protein. AB - The mouse TRPV6 gene is localized on chromosome 6 and extends over 15.66kb. The encoded protein comprises 727 amino acid residues with a calculated relative molecular mass of 83,210Da. TRPV6 is glycosylated and both variants, the glycosylated and the de-glycosylated proteins, are recognized by various polyclonal and monoclonal antibodies, which were raised against TRPV6. Like human TRPV6, mouse TRPV6 binds calmodulin in the presence, but not in the absence of Ca2+. TRPV6 is abundantly expressed in mouse pancreas and placenta, and to a much lesser extend in mouse stomach and kidney. No transcript expression was detected in poly(A)+RNA isolated from heart, brain, intestine, esophagus or aortic endothelial cells. PMID- 12765697 TI - TRPM2 Ca2+ permeable cation channels: from gene to biological function. AB - TRPM2 is a recently identified TRPM family cation channel which is unique among known ion channels in that it contains a C-terminal domain which is homologous to the NUDT9 ADP-ribose hydrolase and possesses intrinsic ADP-ribose hydrolase activity. Here, available information on the TRPM2 gene, transcripts, predicted protein products, and assembled multimeric channels is comprehensively reviewed and synthesized to highlight important areas for future work and provide insight into potential biological function(s) of TRPM2 channels. PMID- 12765698 TI - Role and regulation of TRP channels in neutrophil granulocytes. AB - Members of the transient receptor potential (TRP) family for which mRNA can be demonstrated in neutrophil granulocytes with RT-PCR include TRPC6 (as only "short" TRP), TRPM2, TRPV1, TRPV2, TRPV5 and TRPV6. When these are analyzed in heterologous overexpression experiments, TRPM2 is the only cation channel with characteristic properties that can be used as fingerprint to provide functional evidence for its expression in neutrophil granulocytes. As cells transfected with TRPM2, neutrophil granulocytes display non-selective cation currents and typical channel activity evoked by intracellular ADP-ribose and NAD. Thus, stimulation of TRPM2 is likely to occur after activation of CD38 (producing ADP-ribose) and during the oxidative burst (enhancing the NAD concentration). This novel mode of cation entry regulation may be of particular importance for the response of granulocytes to chemoattractants. TRPV6 is a likely but not exclusive candidate as subunit of the channels mediating store-operated Ca2+ entry (SOCE). Evidence for SOCE in granulocytes has been presented with the fura-2 technique but not with electrophysiological methods although Ca2+-selective store-operated currents can be demonstrated in HL-60 cells, a cell culture model of neutrophil granulocytes. PMID- 12765699 TI - Making sense with TRP channels: store-operated calcium entry and the ion channel Trpm5 in taste receptor cells. AB - The sense of taste plays a critical role in the life and nutritional status of organisms. During the last decade, several molecules involved in taste detection and transduction have been identified, providing a better understanding of the molecular physiology of taste receptor cells. However, a comprehensive catalogue of the taste receptor cell signaling machinery is still unavailable. We have recently described the occurrence of calcium signaling mechanisms in taste receptor cells via apparent store-operated channels and identified Trpm5, a novel candidate taste transduction element belonging to the mammalian family of transient receptor potential channels. Trpm5 is expressed in a tissue-restricted manner, with high levels in gustatory tissue. In taste cells, Trpm5 is co expressed with taste-signaling molecules such as alpha-gustducin, Ggamma(13), phospholipase C beta(2) and inositol 1,4,5-trisphosphate receptor type III. Biophysical studies of Trpm5 heterologously expressed in Xenopus oocytes and mammalian CHO-K1 cells indicate that it functions as a store-operated channel that mediates capacitative calcium entry. The role of store-operated channels and Trpm5 in capacitative calcium entry in taste receptor cells in response to bitter compounds is discussed. PMID- 12765700 TI - Transient receptor potential (TRP) channels as potential drug targets in respiratory disease. AB - Calcium-permeable channels have traditionally been thought of as therapeutic targets in excitable cells. For instance, voltage-operated Ca2+ channels in neurones and smooth muscle cells for neurological and cardiovascular diseases although calcium-permeable channels are also functionally important in electrically non-excitable cells. In the lung, calcium channels play a pivotal role in the activation of all the cell types present, whether resident cells such as airway smooth muscle cells and macrophages or migratory cells such as neutrophils or lymphocytes.Previously, research in this area has been hindered by the lack of obvious molecular identity. More recently, the emergence of the transient receptor potential (TRP) cation family has yielded promising candidates which may underpin the different receptor-operated calcium influx pathways. The challenge now, is to ascribe function to the TRP channels expressed in each cell type as a first step in identifying which TRP channels may be potential drug targets for asthma and chronic obstructive pulmonary disease (COPD) (Fig. 1). PMID- 12765701 TI - Reshaping the carcinogenic risk assessment of medicines: international harmonisation for drug safety, industry/regulator efficiency or both? AB - The most significant institutional entity involved in the harmonisation of drug testing standards worldwide is the International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH), which comprises the three pharmaceutical industry associations and regulatory agencies of the EU, US and Japan. It is often claimed that such harmonisation will both accelerate the development and approval of new drugs and preserve safety standards, if not strengthen safety regimes. Drawing on extensive documentary research and interviews, this paper systematically examines whether the efforts by the ICH to improve industrial and regulatory efficiency by harmonising drug testing requirements is likely to raise, maintain or compromise safety standards in carcinogenic risk assessment of pharmaceuticals. The evidence suggests that, in the field of carcinogenicity testing, the ICH management of international harmonisation of medicines regulation is not achieving simultaneous improvements in safety standards and acceleration of drug development. Rather, the latter is being achieved at the expense of the former. Indeed, the ICH may be converting permissive regulatory practices of the past into new scientific standards for the future. These findings are significant as many expert scientific advisers to drug regulatory agencies seem to have accepted uncritically the conclusions reached by the ICH, which may affect a potential patient population of half a billion and tens of thousands of clinical trials. PMID- 12765702 TI - Physician-patient interaction: a gynecology clinic in Turkey. AB - Evidence for gender differences in physicians' communication with their patients comes primarily from Western countries. Little is known about whether these gender differences would also be observed in Turkey, where there are explicit rules about male-female conduct. The purpose of this study was to observe male and female gynecologists' communication with their patients in a gynecology clinic at a state hospital in Istanbul, Turkey. Four male and three female gynecologists were observed in their interaction with 70 patients over 10 days. The observations were conducted during both the history taking and the actual examination sessions by a woman researcher. The results reported in this paper are based on the extensive field notes taken during the observations. Important differences were revealed in interactions between male vs female gynecologists and their patients. Namely, interactions differed in terms of conversation initiation, communication style, use of technical and colloquial language, frequency of eye contact, patience, and provision of information. Communication characteristics specific to interactions between male gynecologists and their patients included a 'blaming the victim' approach, differential treatment of patients, and underestimation of patients' abilities. Environmental factors that affected physicians' interaction with their patients are reported in conjunction with physicians' use of these external factors to explain the problems they experienced in physician-patient interaction. The discussion focuses on alternative explanations for and future research implications of the observed differences between male and female gynecologists in this setting. PMID- 12765703 TI - Reproductive health indicators for China's rural areas. AB - We report community-based development of reproductive health indicators for China's rural areas. To generate these indicators, we sequenced two participatory techniques known as nominal group process and Delphi survey methodology. Nominal group process entailed grassroots reproductive health workers' generating indicators, followed by refinement and prioritization of these indicators through a consensus-building Delphi process among nationally and internationally known reproductive health experts. Major criteria for the indicators were practicality, feasibility, and measurability within China's rural areas. We explain the importance of establishing these indicators for application in rural China and other developing countries as a complement to the World Health Organization's reproductive health indicators for global monitoring; present the identified indicators; and describe lessons learned from field testing in low-, middle-, and high-income counties of China's countryside. PMID- 12765704 TI - When being alone might be better: neighborhood poverty, social capital, and child mental health. AB - Public health researchers have provided a growing body of evidence on the salutary effects of social capital for individual well being. The importance of these findings for social epidemiology, however, may have precluded so far a full examination of the complex association between neighborhood social processes and the well being of individual residents, including the often acknowledged potential "downside" of social capital. In this study, we examine the association between attachment to community, an indicator of social capital, in a sample of African American parents, and the presence of behavior problems in their preschool children. Participants were recruited from a socioeconomically diverse set of neighborhoods. Attachment to community was assessed using a multi-item scale comprised of two subscales, general sense of community and how well one knew one's neighbors. Results indicated that the association between how well a parent knew her neighbors and the presence of child behavior problems differed depending on the degree of economic impoverishment of the neighborhood. In wealthy neighborhoods, children whose parent reported knowing few of the neighbors had higher levels of internalizing problems such as anxiety and depression compared to those who knew many of their neighbors. In contrast, in poor neighborhoods, children whose parent reported knowing few of the neighbors had lower levels of internalizing problems compared to those who knew many of their neighbors. These results are discussed in terms of furthering the study of the contextual nature of the social capital in explaining community inequalities in mental health among children. PMID- 12765705 TI - Deliberations about deliberative methods: issues in the design and evaluation of public participation processes. AB - A common thread weaving through the current public participation debate is the need for new approaches that emphasize two-way interaction between decision makers and the public as well as deliberation among participants. Increasingly complex decision making processes require a more informed citizenry that has weighed the evidence on the issue, discussed and debated potential decision options and arrived at a mutually agreed upon decision or at least one by which all parties can abide. We explore the recent fascination with deliberative methods for public involvement first by examining their origins within democratic theory, and then by focusing on the experiences with deliberative methods within the health sector. In doing so, we answer the following questions "What are deliberative methods and why have they become so popular? What are their potential contributions to the health sector?" We use this critical review of the literature as the basis for developing general principles that can be used to guide the design and evaluation of public involvement processes for the health care sector in particular. PMID- 12765706 TI - "Love thy neighbour"-it's good for your health: a study of racial homogeneity, mortality and social cohesion in the United States. AB - This paper explores the idea that in societies that experience racial tension, increasing racial heterogeneity will be associated with poorer health outcomes, and this effect will be observable in the health of both the minority and the majority group. Here, the association between mortality and racial homogeneity in the United States is examined. The level of racial homogeneity, indexed by the proportion of blacks in each state of the 50 states in the US, was examined in relation to all-cause mortality, adjusted for age and disaggregated by race and sex. The level of poverty in each state was controlled for in ordinary least squares regression models. The level of racial homogeneity was significantly associated with age adjusted mortality rates for both blacks and whites, accounting for around 30% of the variance in mortality rates in the total population and the white population. Every 1% increase in the percentage of the state population who were black was associated with an increase in the total mortality rate of 5.06 per 100000 and an increase in the white mortality rate of 3.58 per 100000. Based on the data, this suggests, for example, that racial heterogeneity in Mississippi accounts for around 14% of the white mortality rate and in New York and Delaware it accounts for around 7%. These results appear to support the social cohesion thesis that in societies that are intolerant, mortality rates will increase as the proportion of racial or ethnic minorities increase in population. Limitations and explanations for the findings are discussed. PMID- 12765707 TI - Tracing the social gradient in the health of Canadians: primary and secondary determinants. AB - The social gradient in heath refers to the fact that inequalities in population health status are related to inequalities in social status. This study advances and tests a model of the relationships between what we term primary and secondary determinants of the social gradient in health. The primary determinants of health include socioeconomic and demographic indicators. Secondary determinants include stressors, control, self-esteem, social support, and social involvement. Health status is indicated by measures of physical health, self-reported health status, and mental distress. Data are taken from the Canadian National Population Health (NPH) Survey (1994-1995). The study sample consists of 7720 men and 9269 women 15 to over 80 years of age. Using path analysis, we found that higher household income, being retired and growing older are significantly associated with lower stressor levels. Higher stressor levels are associated with lower levels of control, self-esteem, and social support. Higher income Canadians experience greater levels of control and social support, while older Canadians experience lower rates of social support but higher rates of social involvement. Being employed and caring for one's family are positively associated with better physical and self-reported health status. Higher household income, being retired, and aging are associated with better physical health and lower mental distress when accounting for their role in lowering stressor levels and bolstering control, self-esteem, social support, and social involvement. Replicating this study with future samples of the NPH Survey should be of benefit in ascertaining whether the social gradient in Canadians' health status shows signs of declining. PMID- 12765708 TI - Negotiating palliative care expertise in the medical world. AB - This paper explores the relationship between palliative medicine and the wider medical world. It draws on data from a focus group study in which doctors from a range of specialties talked about developing palliative care for patients with heart failure. In outlining views of the organisation of care, participants engaged in a process of negotiation about the roles and expertise of their own, and other, specialties. Our analysis considers the expertise of palliative medicine with reference to its technical and indeterminate components. It shows how these are used to promote and challenge boundaries between medical specialties and with nursing. The boundaries constructed on palliative medicine's technical contribution to care are regarded as particularly coherent within orthodox medicine. In contrast, its indeterminate expertise, represented by the 'holistic' and 'psychosocial' agendas, is potentially compromising in a medical world that prizes science and rationality. We show how the coherence of both kinds of expertise is contested by moves to extend palliative care beyond its traditional temporal (end-of-life) and pathological (cancer) fields of practice. PMID- 12765709 TI - Problematising depression: young people, mental health and suicidal behaviours. AB - The published literature provides strong evidence for connections between mental health issues, such as depression, and suicidal behaviours. However, in spite of this, no investigations to date have explored young people's perceptions of the interconnections between depression, and suicidal behaviours. This article presents discussive analyses of discussions of the contributions of depression to their suicidal behaviours of young people in New Zealand. Two dominant discourses of depression emerged: a medicalised discourse, and a moral discourse. The medicalised discourse was accessible to the majority of participants, and constructed depression as a disease. This discourse prioritised the voices of health professionals and suggested that depression was difficult to resist. The moral discourse was an alternative to the medicalised discourse, and constructed young people who experienced depression and suicidal behaviours as failures. Both discourses were informed by a mechanistic cause-and-effect relationship between depression and suicidal behaviours: attempting suicide was seen as an inevitable outcome of experiencing depression, and suicidal behaviours were inevitably undertaken by young people who were depressed. Resistance to either of these dominant discourses was problematic, and was best articulated during discussions of the stigma associated with mental ill-health and depression. PMID- 12765710 TI - Changing patterns of orphan care due to the HIV epidemic in western Kenya. AB - The HIV/AIDS epidemic has given rise to major demographic changes including an alarming number of orphans in sub-Saharan Africa. The study describes a rural community in western Kenya in which one out of three children below 18 years of age had lost at least one biological parent-and one out of nine had lost both. The main problems these children faced were lack of school fees, food and access to medical care. The high number of orphans has overwhelmed the traditional mechanisms for orphan care, which were based on patrilineal kinship ties. Thus, 28% of the orphans were looked after by culturally "inappropriate" categories such as matrilineal kin or strangers. Furthermore, many of the caretakers were themselves not capable due to ill health or old age. Factors such as poverty, negative attitudes, and traditional funeral customs made the orphans' situation even worse. The authors conclude that though community-based interventions are urgently needed as the most appropriate way to address the issue, the complex, local reality in which cultural factors, kinship ties, and poverty are interwoven needs to be taken into consideration if sustainable solutions are to be found. PMID- 12765711 TI - Children's feeding programs in Atlantic Canada: some Foucauldian theoretical concepts in action. AB - Since 1989 the number of Canadian children depending on food banks has increased by more than 85%. To combat perceived hunger, breakfast and lunch programs have been initiated by localized volunteer efforts. This paper attempts to show the Foucauldian concepts of power, truths, space and time in action in feeding programs in Atlantic Canada. A potential 'relation of docility-utility' is imposed upon children by providers of feeding programs and ultimately the state. The 'power over life' or 'micro-physics of power' is accomplished through procedures that use food, rules, rewards, reinforcements, space, time, and truths. Children voluntarily subject themselves to this relation while reserving the power to resist through acts of defiance or by not attending at all. This ability to exercise one's agency allows for shifting power relations in the social dynamics of feeding programs. The potentially coercive nature of these relationships is embedded in the pleasurable environment generated by the feeding process. PMID- 12765712 TI - Return migration in the context of parental assistance in the AIDS epidemic: the Thai experience. AB - Most persons with AIDS (PWAs) eventually require demanding caregiving. This can prompt changes in living arrangements during the course of the illness. Few studies have attempted to examine the potential links between AIDS and migration from this perspective. The present study uses both direct and indirect approaches to examine the extent of return migration of adults with AIDS in Thailand and explores how this is linked to residence with and care by older aged parents. Methodological challenges and various approaches to the study of this phenomenon are discussed. Despite differences in the nature of information available from our samples and in basic sample characteristics, the findings show a consistent pattern suggestive of extensive return migration among PWAs. The fact that most return migrants die within a few months of their return indicates that they are seeking parental caregiving during the final stages of the illness. The vast majority of PWAs who returned home after becoming ill did so because of their illness, particularly due to a need for care. PMID- 12765713 TI - A shift in the historical trajectory of medical dominance: the case of Medibank and the Australian doctors' lobby. AB - During the twentieth century, episodes of major health policy reform were relatively rare. These rare episodes were pivotal 'critical junctures' in determining the paths taken by modern health systems and in creating a unique health policy arena. A pivotal 'turning point' in Australia came in the form of national health insurance. Introduced in the early 1970s, Medibank (universal, compulsory national health insurance) was the first significant structural health policy change in Australia during the twentieth century. When Medibank health insurance proposals were presented in Australia, political struggles erupted. Government leaders in Australia faced fierce opposition from key players within the health policy arena. Prior to this turning point, one of the key health policy players-the Australian Medical Association (AMA)-had developed a corporate partnership with the non-Labor government. When the Medibank proposal emerged, power structures in the health policy arena were re-aligned. The political role of the AMA shifted from a corporate partner to a pressure group.Examining the political processes surrounding this unique episode of major health policy change helps to illuminate the dual and dynamic nature of the doctors' lobby. Our study aims to demonstrate empirically Day and Klein's proposition that the doctors' lobby operates as a pressure group, rather than as a corporate-style partner, during periods of structural reform in health care [Political Studies, 40 (1992) 462]. This case study of the doctors' lobby during Medibank negotiations represents a rare break in the tradition of ultimate medical professional veto power in health policy decision-making and provides empirical evidence that challenges a widely held perception about an inevitable historical path of medical dominance. PMID- 12765714 TI - Endangering safe motherhood in Mozambique: prenatal care as pregnancy risk. AB - Despite high infant and maternal mortality rates, many Mozambican women with access to prenatal services delay prenatal clinic consultations, limiting opportunity for prevention and treatment of preventable pregnancy complications. Ethnographic research, interviews with health providers and longitudinal pregnancy case studies with 83 women were conducted in Central Mozambique to examine pregnant women's underutilization of clinic-based prenatal services. The study found that pregnancy beliefs and prenatal practices reflect women's attempts to influence reproduction under conditions of vulnerability at multiple levels. Women reported high maternal reproductive morbidity, frequent pregnancy wastage, and immense pressure to bear children throughout their reproductive years. Reproductive vulnerability is intensified by poverty and an intense burden placed on poor, peri-urban women farmers for family subsistence and continuous fertility in a period of economic austerity, land shortages, and increasing social conflict and inequality. In this environment of economic insecurity exacerbated by congested living conditions, women report competing for scarce resources, including male support and income. This vulnerability heightens women's perceptions that they and their unborn infants will be targets of witchcraft or sorcery by jealous neighbors and kin. They respond by hiding pregnancy and delaying prenatal care. Within the context of women's perceived reproductive risks, delayed prenatal care can be seen as a strategy to protect pregnancy from purposeful human and spirit harm. Women mobilized limited resources to acquire prenatal care outside the formal clinic setting. It is concluded that provision of clinical prenatal services is insufficient to reduce reproductive risks for the most socially and economically marginal since it is their vulnerability that prevents women from using available services. Confidential maternity services and social safety nets for greater economic security are recommended. PMID- 12765715 TI - The role of social support and self-esteem in the presence and course of depressive symptoms: a comparison of cancer patients and individuals from the general population. AB - The key focus of this longitudinal study in the Netherlands was to determine the role of social support (i.e. perceived availability of emotional support, lack of received problem-focused emotional support, and negative interactions) and positive and negative self-esteem in depressive symptoms in 475 recently diagnosed cancer patients and 255 individuals without cancer from the general population. Patients and the comparison group were interviewed and filled in a questionnaire at two points in time: 3 months (T1) and 15 months (T2) after diagnosis. The results indicated that social support and self-esteem were weakly to moderately related to each other. Negative self-esteem was more strongly related to all three types of social support, compared to positive self-esteem. Regression analyses showed that social support and self-esteem were independently related to depressive symptoms (concurrently), such that lower levels of social support and self-esteem were strongly associated with higher levels of depressive symptoms. This finding suggests that these two resources supplement each other additively. A longitudinal analysis showed that social support and self-esteem also predicted future levels of depressive symptoms, although the explained variance was much lower than in a cross-sectional analysis. Comparisons between cancer patients and the comparison group generally revealed no significant differences between the two groups in the associations of social support and self esteem with depressive symptoms. The only exception was a lack of problem-focused emotional support. At three months after diagnosis, a lack of this type of support, characterised by reassuring, comforting, problem-solving, and advice, was more strongly related to depressive symptoms in patients than in the comparison group. PMID- 12765737 TI - The efficacy of quetiapine vs haloperidol and placebo: a meta-analytic study of efficacy. AB - INTRODUCTION: Atypical antipsychotics form a new class of treatment for psychotic disorders that offers advantages over conventional antipsychotics, such as haloperidol. Among these advantages is a lower risk of side effects-in particular movement disorders. The atypical antipsychotics that are currently commercially available are clozapine, risperidone, olanzapine, quetiapine, and ziprasidone. The focus of this report is on the efficacy of quetiapine. DATA SOURCES/STUDY SELECTION: A meta-analysis was performed on three placebo- and five haloperidol controlled clinical trials of quetiapine. Efficacy was assessed using the Brief Psychiatric Rating Scale (BPRS), the Positive and Negative Syndrome Scale (PANSS), the Clinical Global Impression (CGI), and the Scale for the Assessment of Negative Symptoms (SANS). In addition, a responder analysis was performed assessing patients who demonstrated a 40% improvement on the BPRS total score. RESULTS: The results showed that quetiapine was significantly (p<0.05) superior to placebo in improving psychotic symptoms. In addition, quetiapine was not significantly different from haloperidol on measures of efficacy measured by BPRS change score, but was superior to haloperidol in terms of response rate using observed case analysis (but not using last observation carried forward analysis). CONCLUSION: This meta-analysis supports the use of quetiapine as a front-line treatment for schizophrenia. PMID- 12765738 TI - Increased developmental deviance and premorbid dysfunction in early onset schizophrenia. AB - Abnormal neurodevelopment and poor premorbid function have been described in schizophrenia. It is unclear whether abnormalities in these domains are increased in patients with early onset schizophrenia (EOS; onset before the 18th birthday) and whether they act to precipitate the earlier onset of the disorder. To address these questions, we collected information based on maternal interviews about the premorbid function of 40 adolescents with recent onset schizophrenia and an equal number of healthy controls using the Developmental Scale Score, the Premorbid Schizoid and Schizotypal Trait Scale (PSST) and Premorbid Adjustment Scale (PAS). Data on the PSST and PAS were also available in 54 patients with adult onset schizophrenia (AOS; onset after the 20th birthday). Compared to healthy controls, EOS patients had (a). delayed speech milestones, difficulties in reading and spelling and greater overall developmental deviance; (b). poor premorbid adjustment in childhood, which became even more deviant in adolescence particularly in boys and (c). more schizophrenia spectrum traits. Both premorbid adjustment and personality traits were more abnormal in patients with increased developmental deviance suggesting the possibility that they represent different manifestations of ongoing abnormalities in developmental processes. EOS patients had more impaired premorbid adjustment in adolescence and schizophrenia spectrum traits compared to AOS cases. Age of onset was related to developmental deviance, premorbid schizophrenia spectrum traits and childhood adjustment in EOS patients only. PMID- 12765739 TI - Patterns of premorbid functioning in first-episode psychosis: initial presentation. AB - Premorbid functioning in first-episode psychosis has been reported to be associated with poorer outcome. We assessed premorbid functioning in a sample of 306 subjects newly admitted to an early-psychosis program. Using cluster analyses, we identified four patterns: stable-good, stable-moderate, deteriorating and poor-deteriorating. Results were that relative to the stable good group, the deteriorating and the poor-deteriorating groups had more negative symptoms, poorer social functioning and some evidence of poorer cognitive functioning. The deteriorating group had increased positive symptoms compared to the stable-good group. These results suggest that prior to the onset of the acute psychosis those who have poor social and interpersonal functioning premorbidly present initially with increased social impairment and negative symptoms compared to those who have better premorbid functioning. PMID- 12765740 TI - Recovery style and outcome in first-episode psychosis. AB - Our study aimed to investigate how recovery styles influence 12-month clinical outcome in first-episode psychosis patients. We hypothesised that patients who use an integrative recovery style would have better outcome than those who seal over. A total of 196 first-episode psychosis patients from the Early Psychosis Intervention Centre (EPPIC) participated in the study. Each patient was interviewed at stabilization of their acute psychotic episode, and then again 12 months later, using an Integration/Sealing-over measure. Our results suggest that recovery style may be a useful predictor of outcome. Integrative patients had better outcome and functioning at 12 months. These results were influenced by diagnosis and, unlike previous studies, recovery style changed over time, suggesting it was not a stable trait. The capacity for recovery style to change challenges the suggestion it is a personality trait and raises the need for psychoeducation and other psychological interventions that could improve outcome. PMID- 12765741 TI - The impact of the duration of untreated psychosis prior to first psychiatric admission on the 15-year outcome in schizophrenia. AB - OBJECTIVE: There is a large amount of evidence to support the hypothesis that the duration of untreated psychosis (DUP) prior to first psychiatric admission adversely affects acute treatment response and short-term outcome in schizophrenia. However, only few prospective studies have attempted to address a possible association between DUP and long-term outcome. METHOD: Fifty-eight DSM III-R schizophrenic patients were assessed at their first psychiatric admission and after a 15-year course of the illness. The 15-year outcome in different domains was compared between patients with different DUPs prior to the first psychiatric admission. RESULTS: A longer DUP was associated with more pronounced negative, positive and general psychopathological symptoms as well as a lower global functioning 15 years after the first psychiatric admission, even after effects of other factors, possibly related to the long-term outcome, were controlled for. CONCLUSIONS: The DUP prior to first psychiatric admission adversely affects the long-term outcome in schizophrenia. The findings underline the importance of establishing health service programs for early detection and treatment of schizophrenic patients with the aim to shorten the DUP and to consequently improve the course and outcome of schizophrenic patients. PMID- 12765742 TI - Sex differences in psychosis: normal or pathological? AB - BACKGROUND: Schizophrenia first appears in adolescence, in boys at an earlier age than girls. The interpretation of this key epidemiological finding crucially depends on whether similar age-related sex differences exist in the expression of associated, subclinical psychosis-like experiences. METHODS: Findings are based on a population sample of 2548 adolescents and young adults aged 17-28. Subjects were assessed with the core psychosis sections on delusions and hallucinations of the Munich-Composite International Diagnostic Interview. RESULTS: The risk of subclinical psychotic experiences was significantly higher for males in the younger half of the cohort (17-21 years), but similar in the older half (22-28 years). CONCLUSIONS: These findings suggest that normal maturational changes in adolescence with differential age of onset in boys and girls cause the expression of psychosis, the extreme of which is schizophrenia. PMID- 12765743 TI - Distinguishing between first-admission schizophreniform disorder and schizophrenia. AB - BACKGROUND: The validity of schizophreniform disorder remains controversial. Past research suggests that cases of schizophreniform disorder may be: (1). atypical cases of affective disorders, (2). cases of schizophrenia in early course, or (3). a heterogeneous group of disorders including a subgroup with benign course and outcome which maintains this diagnosis in the long term. METHOD: We tested the validity of the schizophreniform disorder diagnosis by comparing the socio demographic and baseline clinical characteristics, 24-month course and outcome, and 6- and 24-month research diagnoses of 34 cases initially diagnosed with schizophreniform disorder, and 128 cases with schizophrenia, drawn from a cohort of 628 first-admission patients in the Suffolk County Mental Health Project. RESULTS: Compared to patients with schizophrenia, those with schizophreniform disorder were more likely to remit fully by 6 months and retain this status by 24 months. Only about half of the patients with schizophreniform disorder were re diagnosed with schizophrenia or schizoaffective disorder at 24-month follow-up, 13% were re-diagnosed with affective disorders and 19% retained the diagnosis of schizophreniform disorder. In contrast, 92% of cases with a baseline diagnosis of schizophrenia retained this diagnosis at 24-month follow-up. The findings were similar in comparisons with schizophrenia patients having onset of symptoms within 6 months of hospitalization. CONCLUSIONS: Schizophreniform disorder is a heterogeneous category, which includes a small group with benign psychotic disorders who maintain this diagnosis over at least 24 months. Better delineation of this subgroup has important treatment implications. PMID- 12765744 TI - Cognitive functioning in first episode psychosis: initial presentation. AB - This first part of a longitudinal study examined the initial cognitive performance of 312 individuals who recently presented with a first episode (FE) of psychosis. All attend a comprehensive first episode program. Deficits on a wide range of cognitive tests were observed, suggesting impairment similar to that seen in those with an established schizophrenia illness. There was no evidence to support differences in cognition among the different schizophrenia spectrum diagnostic groups. PMID- 12765745 TI - A comparison of two doses of melperone, an atypical antipsychotic drug, in the treatment of schizophrenia. AB - Melperone at a dose of 300 mg/day has been reported to be as effective as thiothixene and superior to placebo in the treatment of schizophrenia. Limited ability to cause extrapyramidal side effects (EPS) and absence of an effect on plasma prolactin (pPRL) levels suggests that it is an atypical antipsychotic drug. The goal of this pilot study was to determine: (1). the ability of melperone 400 mg/day to produce greater improvement in psychopathology than melperone 100 mg/day; and (2). to compare side effects of these two doses of melperone. Melperone, 100 or 400 mg/day, was administered to 34 acutely hospitalized patients with schizophrenia for 6 weeks in a randomized, double blind manner. Psychopathology, EPS, pPRL levels, and body mass index (BMI) were evaluated at baseline and 6 weeks. Twenty-seven completed the 6-week treatment. A last carried forward analysis revealed no significant difference in the ability of the two doses of melperone to improve psychopathology as measured by the Brief Psychiatric Rating Scale (BPRS)-Total and Positive subscale, the Scale for the Assessment of Negative Symptoms (SANS), the Schedule for Affective Disorders and Schizophrenia-Disorganization subscale, and the Global Assessment Scale (GAS). Treatment with melperone was not associated with exacerbation of EPS, or an increase in pPRL levels or BMI. The Abnormal Involuntary Movement Scale (AIMS) was not significantly changed by treatment with melperone. These results suggest that melperone was equally effective at doses 100 and 400 mg/day, for ameliorating psychopathology and improving overall psychiatric status in patients with schizophrenia. However, the lack of difference and a placebo control group, as well as modest degrees of change in psychopathology, require caution about assuming efficacy of either dose. The lack of significant side effects such as exacerbation of EPS, pPRL elevation, and weight gain indicates melperone is well tolerated. PMID- 12765746 TI - Body mass index and quality of life in community-dwelling patients with schizophrenia. AB - OBJECTIVE: To examine the associations between sociodemographic variables, body weight and quality of life in schizophrenic outpatients. METHODS: Assessments included an interview to obtain sociodemographic data, administration of a Quality of Life questionnaire (the MOS SF-36) and measurement of height and weight. Body mass index was calculated (kg/m(2)). SF-36 subscores were examined for statistical differences based on BMI categories: healthy weight (BMIor=30). Correlations with sociodemographic variables were also examined. RESULTS: Body weight was inversely correlated (level por=7% increase versus initial weight) was also higher with olanzapine (45.7%) than with risperidone (30.6%) and haloperidol (22.4%). Five patients (13.5%) treated with quetiapine had some degree of weight gain according to the UKU scale, although no patient showed a clinically relevant weight gain (>or=7%). Treatment with olanzapine and risperidone were identified as risk factors of weight gain versus haloperidol. The risk of weight gain was higher in women (OR: 4.4), overweight patients (OR: 3.0) and in patients with 3 (or 6) Gal beta 1-->4GlcNAc beta 1-->R structure without participation of other enzymes. The two enzymes differently but cooperatively act on NCAM and the amount of polysialic acid synthesized by both enzymes together is greater than that synthesized by either enzyme alone. The polysialyltransferases are thus important regulators in polysialic acid synthesis and contribute to neural development in the vertebrate. PMID- 12765790 TI - Localization of defined carbohydrate epitopes in bovine polysialylated NCAM. AB - Polysialylated neural cell adhesion molecule (NCAM) was immunoaffinity-purified from the brains of newborn calves. A degree of polymerization of up to 40 was chromatographically determined for released polysialic acid (PSA) chains. For characterization of N-glycan structures and attachment sites, PSA-NCAM was digested with trypsin, and the generated glycopeptides were fractionated by serial immunoaffinity chromatography using immobilized monoclonal antibodies specific for PSA or the HNK1 epitope, i.e., HSO(3)-3GlcA(beta 1-3)Gal(beta 1 4)GlcNAc(beta 1-, yielding PSA-glycopeptides, HNK-glycopeptides and non-PSA/HNK1 (glyco) peptides. Using a combination of enzymatic deglycosylation, peptide fractionation, mass spectrometry and Edman degradation, HNK1-N-glycans could be assigned to glycosylation sites 2, 4, 5 and 6. Non-PSA/HNK1-glycans were assigned to glycosylation site 2, whereas PSA-N-glycans of bovine NCAM had been already previously shown to be restricted to glycosylation sites 5 and 6 (Glycobiology 12 (2002) 47). Respective oligosaccharides were enzymatically released, labeled with 2-aminopyridine and characterized by linkage analysis and mass spectrometry. Carbohydrate chains bearing PSA or the HNK1 epitope comprised mainly fucosylated, partially sulfated diantennary, triantennary or tetraantennary glycans without bisecting GlcNAc or fucosylated diantennary and triantennary species carrying, in part, bisecting GlcNAc residues, respectively. Some N-glycans simultaneously contained both the HNK1-epitope and PSA. Non-PSA/HNK1-glycans exhibited a heterogeneous pattern of partially truncated, mostly diantennary structures with one to three fucose residues, bisecting GlcNAc and/or sulfate residues. In addition, they were demonstrated to carry, to some extent, the Lewis X epitope. When compared with previous data on murine NCAM glycosylation, our results indicate a conservation of structural features and attachment sites for the different types of NCAM N-glycans. PMID- 12765791 TI - Role of oligomannosidic N-glycans in the proliferation, adhesion and signalling of C6 glioblastoma cells. AB - The potential role of glycoprotein N-glycans in the proliferation and adhesion of C6 glioblastoma cells was investigated using a set of N-glycosylation inhibitors (tunicamycin, deoxynojirimycin, castanospermine, deoxymannojirimycin, swainsonine), and traffic (monensin). It was observed that both the proliferative and adhesive properties of C6 cells were dependent upon the expression at the cell surface of glycoproteins with oligomannosidic and hybrid type N-glycans, whereas the absence of N-glycans (tunicamycin) or the presence of glucosyl oligomannosides (deoxynojirimycin and castanospermine) and the absence of glycoproteins at the cell surface (monensin) reduced both the proliferative and adhesive properties of C6 cells. Studies of the classical elements of signalling pathways indicated that the different inhibitors have a low impact on tyrosine phosphorylations and oncogene product expression (except the ras oncogene product), except on phosphorylations on other residues. An endogenous soluble lectin (CSL; J. Neurochem. 49 (1987) 1250), specific for oligomannosidic and hybrid type N-glycans, was present and externalised by the cells through a pinching-off of large intracellular vesicles, a mechanism that was not blocked by monensine; in contrast with the externalisation of its glycoprotein ligands. The inhibitory effect of anti-CSL Fab fragments on adhesion indicates that the polyvalent CSL acts as a bridging molecule for a family of surface glycoproteins expressed at the surface of C6 cells. The inhibitory effect of the same Fab fragments on the proliferation indicates that CSL is a mitogen for these cells, possibly involved in clustering its surface glycoprotein ligands. A mechanism for the loss of contact inhibition is discussed based on the over-expression of CSL ligands in C6 glioblastoma cells relative to normal cells. PMID- 12765792 TI - Two laminin receptors with N-acetylglucosamine-binding specificity. AB - Two glycoproteins, the first, CBP70 which has lectin properties, and the second, cbg72 which is a laminin-1 receptor, have been previously described. We investigated whether cbg72 could have lectin properties and whether CBP70 could have a laminin-receptor function. We observed that CBP70, like cbg72, is a laminin-binding protein. CBP70 interacts with laminin-1 in a carbohydrate dependent fashion, but this interaction could also be a protein-protein interaction. In parallel, we showed that cbg72, as well as CBP70, is a lectin that recognizes glucose and N-acetylglucosamine in a calcium-dependent manner. Moreover, cross-immunoreactivity was observed between these two lectins using their respective antibodies. The resistance of the two lectins, cbg72 and CBP70, to Triton X-100 extraction, suggests that they potentially interact with cytoskeleton elements, since transmembrane proteins that interact with cytoskeleton elements are known to be resistant to such an extraction. PMID- 12765793 TI - Uptake of sialic acid by human erythrocyte. Characterization of a transport system. AB - Upon incubation of human red blood cells (RBC) with [4-9-14C] N-acetylneuraminic acid, the cells incorporated this sugar, as demonstrated by the identification of labelled N-acetylmannosamine in the cytosol, as a result of the action of the sialic acid pyruvate-lyase we discovered previously (Biochimie 84 (2002) 655). The mechanism is saturable and indicates the presence of a limited number of transporter molecules in the RBC membrane. This transport process may have relevance to the desialylation of membrane glycoconjugates which occurs during ageing of erythrocytes. PMID- 12765796 TI - Medico-legal aspects of neurosurgical practice. PMID- 12765795 TI - The power of the mind. PMID- 12765797 TI - Malpractice crisis. PMID- 12765803 TI - Visualization of the eloquent motor system by integration of MEG, functional, and anisotropic diffusion-weighted MRI in functional neuronavigation. AB - BACKGROUND: In this study, we visualized the eloquent motor system including the somatosensory-motor cortex and corticospinal tract on a neuronavigation system, integrating magnetoencephalography (MEG), functional magnetic resonance imaging (fMRI), and anisotropic diffusion-weighted MRI (ADWI). METHODS: Four patients with brain lesions adjacent to the eloquent motor system were studied. Motor evoked responses (MER) by finger-tapping paradigm were acquired with a 1.5-Tesla MR scanner, and somatosensory-evoked magnetic fields (SEF) by median nerve stimulation were measured with a 204-channel MEG system. In the same fMRI examination, ADWI and anatomic three-dimensional T1-weighted imaging (3-D MRI) were obtained. Activated areas of MER, estimated SEF dipoles, and the corticospinal tract on ADWI were coregistered to 3-D MRI, and the combined MR data were transferred to a neuronavigation system (functional neuronavigation). Intraoperative recording of cortical somatosensory-evoked potentials was performed for confirmation of the central sulcus. RESULTS: Combination of fMRI and MEG enabled firm identification of the central sulcus. Functional neuronavigation facilitated extensive tumor resection, having the advantage of sparing the motor cortex and corticospinal tract in all cases. CONCLUSIONS: The proposed functional neuronavigation allows neurosurgeons to perform effective and maximal resection of brain lesions, identifying and sparing eloquent cortical components and their subcortical connections. Potential clinical application of this technique is discussed. PMID- 12765808 TI - Comparison of revision rates following endoscopically versus nonendoscopically placed ventricular shunt catheters. AB - BACKGROUND: Endoscopic placement of ventriculoperitoneal (VP) shunt catheters in pediatric patients has been increasingly used in an attempt to minimize the unacceptably high rates of revision. Although this procedure carries an increased expense, there is currently no evidence to support an improved long-term outcome. This paper compares the rates of revision following ventricular catheter placement for shunted hydrocephalus with and without the use of endoscopy. METHODS: We retrospectively reviewed the records of all pediatric patients who had undergone shunt placement for hydrocephalus between April 1992 and February 1998. All shunts placed before March 1995 were performed without the endoscope; all subsequent shunts were placed endoscopically. The independent effect of endoscopic versus nonendoscopic shunt placement on subsequent shunt failure was analyzed via multivariate proportional hazards regression model. Multiple logistic regression analyses were used to determine the independent effect of endoscopic placement on subsequent etiology of failure (infection, proximal obstruction, distal malfunction) in the 511 failing shunts. RESULTS: There were 447 pediatric patients who underwent a total of 965 shunt placements or revisions. Six hundred and five (63%) catheters were placed with the use of the endoscope. Three hundred and sixty (37.3%) were placed without the use of the endoscope. Neuroendoscopy did not independently affect the risk of subsequent shunt failure [Hazard Ratio (95% Confidence Interval) = 1.08 (0.84-1.41)]. Endoscopic placement independently decreased the odds [Odds Ratio (95% Confidence Interval) = 0.56 (0.32-0.93)] of proximal obstruction, increased the odds of distal malfunction [1.52 (1.02-2.72)], and was not associated with infection [1.42 (0.78-2.61)]. CONCLUSIONS: Endoscopic assisted ventricular catheter placement decreased the odds of proximal obstruction but failed to improve overall shunt survival in this 6 year experience. PMID- 12765806 TI - Randomized, pilot study of intermittent pneumatic compression devices plus dalteparin versus intermittent pneumatic compression devices plus heparin for prevention of venous thromboembolism in patients undergoing craniotomy. AB - BACKGROUND: Unfractionated heparin and the low molecular weight heparin, dalteparin, are used for prophylaxis against venous thromboembolism in patients undergoing craniotomy. These drugs were compared in a randomized, prospective pilot study comparing intermittent pneumatic compression devices plus dalteparin to intermittent pneumatic compression devices plus heparin. METHODS: One hundred patients undergoing craniotomy were randomly allocated to receive perioperative prophylaxis with subcutaneous (SC heparin, 5000 units every 12 hours, or dalteparin, 2,500 units once a day, begun at induction of anesthesia and continued for 7 days or until the patient was ambulating. Entry criteria were age over 18 years, no deep vein thrombosis (DVT) preoperatively as judged by lower limb duplex ultrasound and no clinical evidence of pulmonary embolism preoperatively. Patients with hypersensitivity to heparin, penetrating head injury or who refused informed consent were excluded. Patients underwent a duplex study 1 week after surgery and 1 month clinical follow-up. All patients were treated with lower limb intermittent pneumatic compression devices. RESULTS: There were no differences between groups in age, gender, and risk factors for venous thromboembolism. There were no differences between groups in intraoperative blood loss, transfusion requirements or postoperative platelet counts. Two patients receiving dalteparin developed DVT (one symptomatic and one asymptomatic). No patient treated with heparin developed DVT and no patient in either group developed pulmonary embolism. There were two hemorrhages that did not require repeat craniotomy in patients receiving dalteparin and one that did require surgical evacuation in a patient treated with heparin. Drug was stopped in two patients treated with dalteparin because of thrombocytopenia. None of these differences were statistically significant. CONCLUSION: There was no significant difference in postoperative hemorrhage, venous thromboembolism or thrombocytopenia between heparin and dalteparin. The results suggest that, given the small sample size of this trial, both drugs appear to be safe and the incidence of venous thromboembolism by postoperative screening duplex ultrasound appears to be low when these agents are used in combination with intermittent pneumatic compression devices. PMID- 12765810 TI - Study on therapeutic mechanism and clinical effect of mild hypothermia in patients with severe head injury. AB - BACKGROUND: The therapeutic mechanism and clinical effect of mild hypothermia in patients with severe head injury were studied. METHODS: All 396 patients with severe head injury [Glasgow Coma Scale score (GCS) equal to or less than 8 on admission] were randomly divided into the hypothermic group (198 cases) and the control group (198 cases). Hypothermia was induced within 24 hours of injury. Rewarming began 1 to 7 days (average 62.4 +/- 27.6 h) after the rectal temperature (RT) reached 32.0 to 35.0 degrees C. Meanwhile, the vital signs, intracranial pressure (ICP), blood gas values, blood electrolytes, brain tissue oxygen pressure (P(bt)O2), brain tissue temperature (BT), cerebral blood flow (CBF), and jugular venous oxygen saturation (S(jv)O2) were measured. The rectal temperature of control patients was induced to 36.5 to 37.0 degrees C. According to GOS, the prognosis of the patients was evaluated. RESULTS: In comparison with control group, during mild hypothermia the high level of ICP, hyperglycemia and blood lactic acid significantly decreased (p < 0.05) and cerebral flow improved dominantly. The vital signs, blood gas values, and blood electrolytes did not change significantly. Decreased mortality and good recovery were also found in hypothermia group. CONCLUSIONS: Mild hypothermia is safe and effective for preventing brain damage on patients with severe head injury, as well as reducing mortality and improving the prognosis. It is important to monitor P(bt)O2, BT, CBF, and S(jv)O2 in hypothermic therapy. PMID- 12765815 TI - Lateral posterior fossa venous sinus relationships to surface landmarks. AB - BACKGROUND: Knowing the location of the venous sinuses in the combined lateral posterior fossa and lateral cranial base approach is important to prevent their inadvertent injury. The identification of surface landmarks related to these structures is useful in planning such surgical approaches. METHODS: Twelve injected adult cadaver specimens and 10 dried skulls were used to study the relationship of the venous sinuses to various surface anatomic structures. RESULTS: The asterion was not clearly seen in 60% of the studied cadaver sides. The asterion was always clearly seen in the dry skull preparations. The upper margin of the superior nuchal line was found to range from 1.5 mm to 14 mm inferior to the lower margin of the lateral transverse sinus. In 85% of our specimens, the mastoid groove was found to completely overlie the sigmoid sinus. CONCLUSIONS: The asterion was found to be variable in its anatomic relations to other identifiable structures. This variability in relation to other posterior fossa bony landmarks limits its overall usefulness as a consistently stable marker for intracranial structures. The first and most superolateral burr hole for lateral posterior fossa procedures can be safely placed 1 cm below the superior nuchal line and 1 cm medial to the top of the mastoid groove. A burr hole in this location will avoid the transverse and sigmoid sinuses, as well as the transverse-sigmoid junction, yet will be high enough and lateral enough to provide easy exposure of these venous sinuses for all lateral posterior fossa procedures. PMID- 12765811 TI - Growth potential of prolactinomas in men: is it really different from women? AB - BACKGROUND: In comparison to prolactinomas in women, those in men are usually large tumors and have been suggested to have a higher proliferation activity. To elucidate the reason for the predominance of large tumors in men, we investigated the gender-related difference of prolactinomas including growth potential. METHODS: We conducted a retrospective study comparing unselected 16 men and 27 women with prolactinoma concerning clinical and endocrine findings, tumor volume, and the mean Ki-67 (MIB-1) positive cell index (PCI). RESULTS: Prolactinomas in men were larger in size (p = 0.0090), higher in serum prolactin level (p = 0.0359), and exhibited higher PCI (p = 0.0138) than those in women. However, the three factors-tumor volume, prolactin level, and the PCI-showed positive correlations with each other. There was no gender-related difference in the PCI when small tumors (<1.0 cm3) were excluded. CONCLUSION: High PCI is associated with large tumors in both men and women. Because large tumors predominate in men, this is in fact consistent with the view that the biologic behavior of prolactinomas is different between the sexes. PMID- 12765817 TI - The effects of hydrogen peroxide on brain and brain tumors. AB - BACKGROUND: Hydrogen peroxide (HP) is routinely used during neurosurgical procedures to augment hemostasis after intracranial tissue resection. Elsewhere in the body, HP is used to kill resection margin tumor cells; in vitro studies support these clinical uses. The effects of HP on brain and brain tumors have not been evaluated. In this study, the in vitro and in vivo effects of HP on both rat and human brain parenchyma and brain tumors were examined. METHODS: Antitumor activity of varied concentrations of HP (0-30%) on cultured primary and metastatic brain tumors (n = 13) was compared with the effects of various concentrations of ethanol (0-50%). Studies were performed in rats to characterize HP-induced tissue changes that occurred when HP-soaked pledgets were placed on the arachnoid surface and along resection margins (n = 5). Additionally, the effect of HP on human brain along tumor resection cavities was investigated (n = 10). RESULTS: While HP demonstrated concentration-dependent tumoricidal effects in vitro, similar to results achieved with ethanol, HP caused significant injury to arachnoid and stroma with neuronal and glial injury to a depth of 1 mm in rats. Three percent HP-soaked pledgets placed in resection cavities of excised brain tumors induced similar injury in human brain. CONCLUSION: HP irreversibly damages mesothelial and neural tissue. Although HP appears to have tumoricidal effects in vitro, it should be used with caution in humans because of risks of collateral injury to surrounding normal brain. HP may prove most beneficial for discrete lesions, such as pituitary tumors and metastases. PMID- 12765819 TI - Spinal subdural hematoma: a sequela of a ruptured intracranial aneurysm? AB - BACKGROUND: A case of spinal subdural hematoma (SSDH) following subarachnoid hemorrhage (SAH) because of a ruptured internal carotid aneurysm is described. Such a case has never been reported. CASE DESCRIPTION: A 52-year-old woman underwent a craniotomy for a ruptured internal carotid aneurysm. A computed tomography scan showed that SAH existed predominantly in the posterior fossa and subdural hematoma beneath the cerebellar tentorium. Intrathecal administration of urokinase, IV administration of fasudil hydrochloride, and continuous cerebrospinal fluid (CSF) evacuation via cisternal drainage were performed as prophylactic treatments for vasospasm. On the sixth postoperative day, the patient complained of severe lower back and buttock pain. Magnetic resonance imaging showed a subdural hematoma in the lumbosacral region. Although the mass effect was extensive, the patient showed no neurologic symptoms other than the sciatica. She was treated conservatively. The hematoma dissolved gradually and had diminished completely 15 weeks later. Her pain gradually subsided, and she was discharged 7 weeks later without any neurologic deficit. CONCLUSION: Although the exact mechanism of SSDH in this case is unclear, we speculate that this SSDH was a hematoma that migrated from the intracranial subdural space. Low CSF pressure because of continuous drainage and intrathecal thrombolytic therapy may have played an important role in the migration of the hematoma through the spinal canal. It is important to recognize the SSDH as a possible complication of the SAH accompanied with intracranial subdural hematoma. PMID- 12765821 TI - Angioplasty and coiling of ruptured aneurysm with symptomatic vasospasm: technical case report. AB - BACKGROUND: Treating a ruptured cerebral aneurysm during symptomatic vasospasm is very difficult. We describe the successful endovascular treatment of such a case and discuss its efficacy. CASE PRESENTATION: A 34-year-old man had a sudden onset of severe headache. One week later, he was referred to our institute with gradually progressing right hemiparesis and global aphasia. Cerebral angiography demonstrated severe vasospasm of the left internal carotid artery system and an anterior communicating artery aneurysm. With the patient under general anesthesia, 90% occlusion of the aneurysm was achieved with detachable coils after successful dilatation of the spastic vessels. The patient had an uneventful postoperative course and his neurologic symptoms were improved. Seven months after the endovascular treatment, the enlarged neck remnant of the aneurysm was successfully clipped without difficulty. CONCLUSION: The simultaneous treatment of a ruptured aneurysm and vasospasm with percutaneous transluminal angioplasty and coils can produce a better outcome for the patient. PMID- 12765823 TI - Amyloidoma of the brachial plexus. AB - BACKGROUND: Amyloidomas of the peripheral nervous system are rare lesions. Most commonly, they involve the gasserian ganglion and the branches of the fifth cranial nerve. No association with systemic amyloidosis has been reported. CASE DESCRIPTION: We describe an amyloidoma of the lower trunk of the right brachial plexus. At the age of 34 years, this 71-year-old female had undergone radical right mastectomy for breast cancer with axillary lymph node dissection followed by radiotherapy. On admission, she presented with burning pain to the right hand and mild motor deficit to the ulnar-innervated intrinsic hand muscles. A palpable lesion was found in the supraclavicular region. On surgical inspection, the lesion appeared to originate from the lower trunk of the right brachial plexus. The middle and upper trunks were dislocated. Histologically, fibrous connective tissue embedded small nerve bundles featuring perineurial and endoneurial fibrosis as well as amyloid. Amyloid featured immunoreactivity for both lambda and kappa chains. DISCUSSION: Localized amyloidoma of brachial plexus has never been reported. Because of compressive rather than infiltrative growth of the present lesion, a conservative surgery was achieved. Our immunohistochemical findings indicated that peripheral nerve amyloidomas are not, by definition, monoclonal in nature. PMID- 12765825 TI - Third ventricular chordoid glioma: case report and review of the literature. AB - BACKGROUND: Chordoid glioma of the third ventricle is a rare type of brain tumor that was recently characterized as a novel tumor entity. We present a case and review of the literature. CASE REPORT: A 49-year-old woman presented with progressive headache, memory impairment and urinary incontinence. MRI showed a large well-circumscribed tumor in the third ventricle. The tumor was partially removed via a trans-lamina terminalis approach. The histologic findings indicated chordoid glioma. Residual tumor was treated by stereotactic radiosurgery and showed no regrowth at 2-year follow-up. CONCLUSIONS: The ideal therapy is total removal of the tumor. However, according to the literature, total removal of the tumor carries a high risk because of its location, and conventional radiation therapy has little effect on the residual tumor. On the other hand, stereotactic radiosurgery appears more promising, and to date, no regrowth has been reported after gamma-knife therapy. PMID- 12765826 TI - Primary epithelioid hemangioendothelioma originating in the lower petroclival region: case report. AB - BACKGROUND: We report the first case of primary epithelioid hemangioendothelioma (EH) originating in the lower petroclival region. CASE DESCRIPTION: A 55-year-old female presented with a 45-year history of subclinical atrophy on the right side of her tongue and a 15-year history of hoarseness. Neuroimaging revealed an expansile, homogeneously enhanced intraosseous mass with bony shell and honeycomb configuration in the petroclival region. A right far lateral transcondylar approach was utilized for subtotal removal of the tumor, which demonstrated high vascularity in the petroclival bone. Histopathological and immunohistochemical examination confirmed the diagnosis of EH. Because of the intermediate malignancy, adjuvant gamma knife radiotherapy was performed for the residual mass 5 months after surgery. CONCLUSION: EH rarely occurs in the skull base region. The appearance of surrounding bony structure and rich vascularity are important findings for the differential diagnosis. A far lateral transcondylar approach provides sufficient exposure of the jugular tubercle and lower clivus. Additional radiotherapy may be recommended for residual tumors with a high MIB-1 labeling index. PMID- 12765828 TI - To the Robin Hood era...and back. PMID- 12765829 TI - Neurosurgeons, work stoppages, and the art of negotiation. PMID- 12765830 TI - Acetate C-C bond formation and decomposition in the anaerobic world: the structure of a central enzyme and its key active-site metal cluster. AB - The structure of carbon monoxide dehydrogenase/acetyl-coenzyme A synthase (CODH/ACS), a central enzyme in the anaerobic metabolism of acetyl-coenzyme A (acetyl-CoA), has been solved to a resolution of 2.2A. The active-site metal cluster responsible for catalyzing acetyl C-C bond synthesis and cleavage, designated the A center, was identified as an Fe(4)S(4) iron sulfur cluster with one of its cysteine thiolates acting as a bridge to an adjacent binuclear metal site. Nickel was found at one position in the binuclear site and the other metal was indicated to be copper - a surprising result, implying a previously unrecognized role for copper. Details of the A center provided new insight into the unusual organometallic mechanism of acetyl C-C bond formation and cleavage, with substantial conformational changes indicated for binding of the large methylcorrinoid protein substrate, and a unique intramolecular channel acting to contain carbon monoxide within the protein and transfer it to the site needed for acetyl-CoA synthesis. PMID- 12765831 TI - HEPN: a common domain in bacterial drug resistance and human neurodegenerative proteins. AB - A novel domain - HEPN (higher eukarytoes and prokaryotes nucleotide-binding domain) - found in several bacterial species is also present in the human protein, sacsin, a chaperonin implicated in an early-onset neurodegenerative disease. The distant structural similarity suggests that this domain might be involved in nucleotide binding. PMID- 12765832 TI - The STIR-domain superfamily in signal transduction, development and immunity. AB - We have identified a conserved sequence segment in transmembrane receptors (including SEFs, IL17Rs) and soluble factors (including CIKS/ACT1) in eukaryotes and bacteria - the SEFIR domain. This sequence domain is part of the new STIR domain superfamily comprising also the TIR domain known to mediate TIR-TIR homotypic interactions. In TOLL/IL1R-like pathways, the cytoplasmically localized TIR domain of a receptor and the TIR domain of a soluble adaptor interact physically and activate signalling. The similarity between the SEFIR and TIR domains involves the conserved boxes 1 and 2 of the TIR domain that are implicated in homotypic dimerization, but there is no sequence similarity between SEFIR domains and the TIR sequence box 3. By analogy, we suggest that SEFIR domain proteins function as signalling components of Toll/IL-1R-similar pathways and that their SEFIR domain mediates physical protein-protein interactions between pathway components. PMID- 12765833 TI - Amidase domains from bacterial and phage autolysins define a family of gamma-D,L glutamate-specific amidohydrolases. AB - Several phage-encoded peptidoglycan hydrolases have been found to share a conserved amidase domain with a variety of bacterial autolysins (N-acetylmuramoyl L-alanine amidases), bacterial and eukaryotic glutathionylspermidine amidases, gamma-D-glutamyl-L-diamino acid endopeptidase and NLP/P60 family proteins. All these proteins contain conserved cysteine and histidine residues and hydrolyze gamma-glutamyl-containing substrates. These cysteine residues have been shown to be essential for activity of several of these amidases and their thiol groups apparently function as the nucleophiles in the catalytic mechanisms of all enzymes containing this domain. The CHAP (cysteine, histidine-dependent amidohydrolases/peptidases) superfamily includes a variety of previously uncharacterized proteins, including the tail assembly protein K of phage lambda. Some members of this superfamily are important surface antigens in pathogenic bacteria and might represent drug and/or vaccine targets. PMID- 12765834 TI - The CHAP domain: a large family of amidases including GSP amidase and peptidoglycan hydrolases. AB - Cleavage of peptidoglycan plays an important role in bacterial cell division, cell growth and cell lysis. Here, we reveal that several known peptidoglycan amidases fall into a family, which includes many proteins of previously unknown function. The family includes two different peptidoglycan cleavage activities: L muramoyl-L-alanine amidase and D-alanyl-glycyl endopeptidase activity. The family includes the amidase portion of the bifunctional glutathionylspermidine synthase/amidase enzyme from bacteria and pathogenic trypanosomes. The glutathionylspermidine synthase is thought to be a key component of the alternative pathway in trypanosomes for protection from oxygen-radical damage and has been proposed as a potential drug target. The CHAP (cysteine, histidine dependent amidohydrolases/peptidases) domain is often found in association with other domains that cleave peptidoglycan. The large number of multifunctional hydrolases suggests that they might act in a cooperative manner to cleave specialized substrates. PMID- 12765835 TI - Regulation of heptaspanning-membrane-receptor function by dimerization and clustering. AB - G-protein-coupled receptors form homomers and heteromers; agonist-induced conformational changes within interacting receptors of the oligomer modify their pharmacology, signalling and/or trafficking. When these receptors are activated, the oligomers rearrange and cluster and a novel mechanism of receptor-operation regulation by oligomer intercommunication is possible. This intercommunication would be assisted by components of the plasma membrane and by scaffolding proteins. Receptor cross-sensitization, cross-desensitization and novel, integrated receptor responses can then develop between oligomeric receptor complexes of the cluster without direct contact between them. This concept gives a new perspective to the understanding of neurotransmission and neuronal plasticity. PMID- 12765836 TI - Standardization of protocols in cDNA microarray analysis. AB - Systematic variations can occur at various steps of a cDNA microarray experiment and affect the measurement of gene expression levels. Accepted standards integrated into every cDNA microarray analysis can assess these variabilities and aid the interpretation of cDNA microarray experiments from different sources. A universally applicable approach to evaluate parameters such as input and output ratios, signal linearity, hybridization specificity and consistency across an array, as well as normalization strategies, is the utilization of exogenous control genes as spike-in and negative controls. We suggest that the use of such control sets, together with a sufficient number of experimental repeats, in-depth statistical analysis and thorough data validation should be made mandatory for the publication of cDNA microarray data. PMID- 12765837 TI - Metabolic pathways in the post-genome era. AB - Metabolic pathways are a central paradigm in biology. Historically, they have been defined on the basis of their step-by-step discovery. However, the genome scale metabolic networks now being reconstructed from annotation of genome sequences demand new network-based definitions of pathways to facilitate analysis of their capabilities and functions, such as metabolic versatility and robustness, and optimal growth rates. This demand has led to the development of a new mathematically based analysis of complex, metabolic networks that enumerates all their unique pathways that take into account all requirements for cofactors and byproducts. Applications include the design of engineered biological systems, the generation of testable hypotheses regarding network structure and function, and the elucidation of properties that can not be described by simple descriptions of individual components (such as product yield, network robustness, correlated reactions and predictions of minimal media). Recently, these properties have also been studied in genome-scale networks. Thus, network-based pathways are emerging as an important paradigm for analysis of biological systems. PMID- 12765838 TI - Insights into the decoding mechanism from recent ribosome structures. AB - During the decoding process, tRNA selection by the ribosome is far more accurate than expected from codon-anticodon pairing. Antibiotics such as streptomycin and paromomycin have long been known to increase the error rate of translation, and many mutations that increase or lower accuracy have been characterized. Recent crystal structures show that the specific recognition of base-pairing geometry leads to a closure of the domains of the small subunit around cognate tRNA. This domain closure is likely to trigger subsequent steps in tRNA selection. Many antibiotics and mutations act by making the domain closure more or less favourable. In conjunction with recent cryoelectron microscopy structures of the ribosome, a comprehensive structural understanding of the decoding process is beginning to emerge. PMID- 12765839 TI - Does Rap1 deserve a bad Rap? AB - The Ras superfamily of small G proteins is remarkable for both its diversity and physiological functions. One member, Rap1, has been implicated in a particularly wide range of biological processes, from cell proliferation and differentiation to cell adhesion. But the diversity of Rap1 has lead to contradictory reports of its effects. Originally identified as an antagonist of Ras-induced transformation, Rap1 can oppose other actions of Ras including regulation of cell growth and differentiation, integrin-dependent responses and synaptic plasticity. Furthermore, recent evidence confirms that Rap1, like Ras, can activate the MAP kinase cascade (ERK) in several cell types. These diverse functions of Rap1 underscore that the activation and action of Rap1 are regulated by complex factors that are cell-type specific. PMID- 12765840 TI - mtDNA depletion myopathy: elucidation of the tissue specificity in the mitochondrial thymidine kinase (TK2) deficiency. AB - Decreased mitochondrial thymidine kinase (TK2) activity is associated with mitochondrial DNA (mtDNA) depletion and respiratory chain dysfunction and is manifested by isolated, fatal skeletal myopathy. Other tissues such as liver, brain, heart, and skin remain unaffected throughout the patients' life. In order to elucidate the mechanism of tissue specificity in the disease we have investigated the expression of the mitochondrial deoxynucleotide carrier, the mtDNA content and the activity of TK2 in mitochondria of various tissues. Our results suggest that low basal TK2 activity combined with a high requirement for mitochondrial encoded proteins in muscle predispose this tissue to the devastating effect of TK2 deficiency. PMID- 12765841 TI - Infantile hypermethioninemia and hyperhomocysteinemia due to high methionine intake: a diagnostic trap. AB - Studies were carried out to identify the cause of combined severe hypermethioninemia and moderate hyperhomocysteinemia in a cluster of 10 infants ascertained between 1999 and early 2001. Although several were thought initially to have cystathionine beta-synthase (CBS) deficiency and treated accordingly, CBS deficiency and other known genetic causes of hypermethioninemia were ruled out by assay of CBS activity in fibroblasts of four patients and by assays of plasma cystathionine and S-adenosylmethionine. Retrospective data on dietary methionine intakes and plasma concentrations of methionine and related metabolites established that the hypermethioninemia in nine of the 10 babies was related to ingestion of an infant protein hydrolysate formula, the methionine content of which had been increased from May 1998 to February 2001. The formula in question has now been reformulated and is no longer available. The 10th infant manifested similar metabolic abnormalities while receiving TPN containing excessive methionine. Brain MRI abnormalities indicative of cerebral edema, most marked in the cerebral cortex and posterior brainstem, occurred in two patients near times of extreme hypermethioninemia. Metabolic and MRI abnormalities resolved when the methionine intake decreased. A third infant had a normal MRI 1 day after the formula was changed. The possible relationship between extreme hypermethioninemia and cerebral edema is discussed and a working hypothesis offered to explain the relative sensitivity of the inferior colliculi, based upon the facts that this is the region most active in glucose utilization and that Na(+),K(+)-ATPase is inhibited by methionine and related metabolites. PMID- 12765842 TI - Molecular characterization of phenylketonuria in South Brazil. AB - Phenylketonuria (PKU) is an autosomal recessive disorder due to phenylalanine hydroxylase (PAH) deficiency. The PAH gene, located at 12q22-q24.1, includes about 90kb and contains 13 exons. To date, more than 420 different alterations have been identified in the PAH gene. To determine the nature and frequency of PAH mutations in PKU patients from South Brazil, mutation analysis was performed on genomic DNA from 23 unrelated PKU patients. The 13 exons and flanking regions of the PAH gene were amplified by PCR and the amplicons were analyzed by single strand conformation polymorphism (SSCP). Amplicons that showed abnormal migration patterns were analyzed by restriction endonuclease digestion and/or sequencing. Twenty-two previously reported mutations were identified including R261X, R408W, IVS2nt5g-->c, R261Q, and V388M. Polymorphisms were observed in 48.8% of the PKU patients, the most frequent being IVS2nt19t-->c, V245V, and IVS12nt-35c-->t. In addition, two novel sequence variants were identified: 1378g-->t in the 3(') untranslated region in exon 13 which may be disease-causing and an intron 12 polymorphism, IVS12nt-15t-->c. The mutation spectrum in the patients from Southern Brazil differed from that observed in patients from other Latin American countries and further defined the molecular heterogeneity of this disease. PMID- 12765843 TI - Expression of human factor VIII under control of the platelet-specific alphaIIb promoter in megakaryocytic cell line as well as storage together with VWF. AB - Hemophilia A, which results in defective or deficient factor VIII (FVIII) protein, is one of the genetic diseases that has been addressed through gene therapy trials. FVIII synthesis does not occur in normal megakaryocytes. In hemophilia patients who have inhibitors to FVIII activity, megakaryocytes could be a protected site of FVIII synthesis and subsequent release. Since von Willebrand factor (VWF) is a carrier protein for FVIII, we hypothesize that by directing FVIII synthesis to megakaryocytes, it would traffick together with VWF to storage in megakaryocyte alpha-granules and the platelets derived from these cells. Such synthesis would establish a protected, releasable alpha-granule pool of FVIII together with VWF. When platelets are activated in a region of local vascular damage, FVIII and VWF could potentially be released together to provide improved local hemostatic effectiveness. To direct FVIII expression to the megakaryocyte lineage, we designed a FVIII expression cassette where the human B domain deleted FVIII cDNA was placed under the control of the megakaryocytic/platelet-specific glycoprotein IIb (alphaIIb) promoter. We demonstrated by means of a functional FVIII activity assay that the biosynthesis of FVIII occurred normally in Dami cells transfected with FVIII. FVIII production was higher when driven by the alphaIIb promoter compared to the CMV promoter, and was increased about 8-fold following PMA treatment of the transfected Dami cells. Immunofluorescence staining of the transfected cells showed that FVIII stored together with VWF in the granules. The data indicate that the megakaryocytic compartment of hematopoietic cells may represent a potential target of gene therapy for hemophilia A-especially in those patients who have developed inhibitors to plasma FVIII. PMID- 12765844 TI - Analysis of clones from a human cartilage cDNA library provides insight into chondrocyte gene expression and identifies novel candidate genes for the osteochondrodysplasias. AB - To begin to define the gene expression pattern in fetal cartilage and to identify uncharacterized candidate genes for the osteochondrodysplasias, we analyzed clones from a fetal cartilage cDNA library. Sequence analysis of 420 cDNA clones identified 210 clones derived from established genes but, for many of them, expression in cartilage had not been previously reported. Among the established genes were 14 genes known to produce skeletal abnormalities in either humans or mice when mutated. Thirty-two uncharacterized genes and their respective chromosomal positions were also identified. To further understand the expression profile of these genes in fetal cartilage, we constructed a cDNA microarray utilizing the clones. The microarray was used to determine which genes had higher expression in cartilage as compared with dedifferentiated, cultured chondrocytes. Many of the established genes, as well as five of the uncharacterized genes, had increased expression in cartilage, suggesting an important role for these genes in the differentiated state of chondrocytes. These data provide new candidate genes for the osteochondrodysplasias and demonstrate the usefulness of cartilage cDNA microarrays in expanding our understanding of the complexity of fetal cartilage gene expression. PMID- 12765845 TI - Systematic search for single nucleotide polymorphisms in the 5' flanking region of the human phosphodiesterase 3B gene: absence of evidence for major effects of identified polymorphisms on susceptibility to Japanese type 2 diabetes. AB - The activation of phosphodiesterase 3B (PDE3B) reduces free fatty acid output from adipocytes. A reduced PDE3B gene expression could lead to insulin resistance. To determine whether there are polymorphisms associated with type 2 diabetes in PDE3B gene promoter, this 5(') flanking region was isolated. The transcription initiation site was located 206bp upstream from the translation start site. Sequences of 2kb of the 5(') flanking region for 24 type 2 diabetic Japanese subjects were initially analyzed using PCR direct sequencing, and the regions including the identified polymorphisms were then examined. In 98 controls and 98 type 2 diabetic subjects, -1947T>C, -567G>A, -465G>T, -458T>C, and -1727_ 1726insTCAATT were found. Only -465G>T and this insertion had more than 5% frequencies. Since a complete linkage disequilibrium existed between them, 465G>T was further analyzed, along with a previously identified +1389G>A in the coding region, in a total of 200 controls and 207 type 2 diabetic subjects. These allele frequencies were not significantly different between these two groups (controls vs. cases; -465G>T, 12.0% vs. 10.1%, P=0.435; +1389G>A, 30.3% vs. 33.3%, P=0.408). These genotype distributions were not significantly different between these two groups. The T/T genotype at -465 was rare although this frequency could be higher in type 2 diabetes (4/207 subjects) than controls (0/200 subjects). The linkage disequilibrium existed between -465G>T and +1389G>A, and the estimated haplotype frequencies defined by these SNPs were not significantly different between the cases and controls. Thus, the identified polymorphisms are unlikely to have major effects on susceptibility to Japanese type 2 diabetes. PMID- 12765846 TI - Impact of the Pro12Ala polymorphism of the PPAR-gamma2 gene on serum triacylglycerol response to n-3 fatty acid supplementation. AB - Serum lipid responses to dietary modification are partly determined by genetic factors. The objective of the present study was to investigate the influence of the Pro12Ala polymorphism of the peroxisome proliferator-activated receptor gamma2 (PPAR-gamma2) gene on serum lipid and lipoprotein responses to n-3 fatty acid supplementation. A total of 76 men and 74 women (age 49+/-8 years, body mass index 26.5+/-3.0 kg/m(2)) participated in a controlled multi-center study. Subjects were randomly assigned to consume either fish oil supplements (3.6g n-3 fatty acids/day containing 2.4 g of EPA and DHA) or placebo capsules containing olive oil for 3 months. At baseline, the Pro12Ala polymorphism was not associated with serum total and lipoprotein lipid concentrations or lipoprotein lipase activity in the fasting state. After the 3-month study period, carriers of the Ala12 allele presented a greater decrease in serum triacylglycerol concentration in response to n-3 fatty acid supplementation than did subjects with the Pro12Pro genotype when the total dietary fat intake was below 37 E% (p=0.003) or the intake of saturated fatty acids was below 10 E% (p=0.006). Changes in serum total cholesterol, serum LDL cholesterol and HDL cholesterol concentrations were similar among the genotypes in the n-3 fatty acid supplementation group and in the placebo group. In conclusion, the Pro12Ala polymorphism of the PPAR-gamma2 gene may modify the inter-individual variability in serum triacylglycerol response to n-3 fatty acid supplementation. PMID- 12765847 TI - Association of a F479L variant in the cytosolic phospholipase A2 gene (PLA2G4A) with decreased glucose turnover and oxidation rates in Pima Indians. AB - Phospholipase A2, Group IVA (PLA2G4A) belongs to the class of cytosolic calcium dependent phospholipases (cPLA2s) that preferentially cleave arachidonic acid (AA) from membrane glycerophospholipids. AA and AA metabolites play key roles in glucose disposal and insulin secretion. PLA2G4A is located on Chromosome 1q, where a number of groups have reported linkage to type 2 diabetes mellitus. We have screened the PLA2G4A gene and identified a C-->G variant, which predicts a phenylalanine to leucine substitution. In logistic regression analyses adjusted for age, sex, ethnicity, and birth year, we found a trend toward association between this SNP and diabetes [OR=1.53 (0.97-2.40); p=0.06]. Individuals with the variant genotype had lower mean basal endogenous glucose output (1.8+/-0.03 vs. 1.9+/-0.01 mg/kgEMBS/min; p=0.04) and lower mean basal glucose oxidation (1.2+/ 0.11 vs. 1.4+/-0.03 mg/kgEMBS/min; p=0.005) compared to individuals with the wild type genotype. During a low dose insulin infusion, non-diabetic individuals with the variant genotype had a lower mean glucose oxidation (1.9+/-0.11 vs. 2.0+/ 0.03 mg/kgEMBS/min; p=0.04) and total glucose turnover rate (2.5+/-0.22 vs. 2.6+/ 0.06 mg/kgEMBS/min; p=0.01) compared to subjects with the wild-type genotype. In addition, under basal conditions, individuals with the variant genotype had a higher mean lipid oxidation rate compared to individuals with the wild-type genotype (0.77+/-0.25 vs. 0.67+/-0.23 mg/kgEMBS/min; p=0.02). These results provide evidence supporting a role for the eicosanoid biosynthesis pathway in type 2 diabetes mellitus pathophysiology. PMID- 12765848 TI - "Loss of function" mutations in the cationic trypsinogen gene (PRSS1) may act as a protective factor against pancreatitis. AB - Several genetic factors have been well known to predispose one to chronic pancreatitis (CP). However, little is known about the genetic factors that may provide a protective effect against the disease. Having found a nonsense mutation (c.111C>A; Y37X) and a splicing mutation (IVS2+1G>A) in the cationic trypsinogen gene (protease, serine, 1; PRSS1) in alcoholics without the development of CP, but not in alcoholics with CP and patients with hereditary or idiopathic CP, we propose that while "gain of function" mutations in the PRSS1 gene predispose one to pancreatitis, "loss of function" mutations in the gene may protect one against the disease. PMID- 12765849 TI - Add-on rTMS for treatment of depression: a pilot study using stereotaxic coil navigation according to PET data. AB - OBJECTIVE: Repetitive transcranial magnetic stimulation (rTMS) is regarded as a potentially new tool to treat depression. In a double-blind, randomized, sham controlled pilot study we investigated the efficacy of neuronavigated rTMS, guided according to the prefrontal metabolic state determined by positron emission tomography (PET). METHODS: 25 patients with major depression were included. Prior to rTMS, PET scans were obtained. For the real stimulation condition, the dorsolateral prefrontal cortex (DLPFC) with lower metabolic activity compared to the contralateral hemisphere was selected, if detected by prior PET. Stimulation parameters were 15 Hz, 110% motor threshold (MT), 3000 stimuli/day, for 10 days. A neuronavigational system was used to place the magnetic coil above each individuals' selected cortical region (real condition: DLPFC, sham: midline parieto-occipital, intensity 90% of MT). RTMS was administered add-on to medication. Depression-related symptoms were rated with Beck's, Hamilton's (HAM-D), and Montgomery-Asberg's (MADRS) depression rating scales. RESULTS: Real stimulation improved depression according to HAM-D and MADRS moderately but significantly better compared to sham at the end of the stimulation sessions. In the real condition, four out of 13 patients responded with a mean improvement in HAM-D and/or MADRS of at least 50%, whereas none responded to sham. Antidepressant effects of stimulation of the relatively hypometabolic DLPFC were comparable to stimulation in absence of metabolic differences. CONCLUSIONS: A moderate improvement of depressive symptoms after rTMS was observed. Our preliminary data show that stimulation of prefrontal hypometabolism may not be advantageous to stimulation irrespective of the metabolic state. PMID- 12765850 TI - Safety aspects of chronic low-frequency transcranial magnetic stimulation based on localized proton magnetic resonance spectroscopy and histology of the rat brain. AB - Because repetitive transcranial magnetic stimulation (rTMS) is capable of inducing lasting alterations of cortical excitability, it represents a promising therapeutic tool in several neuropsychiatric disorders. However, rTMS, especially when applied chronically, may cause harmful effects in the stimulated tissue. To study the safety of chronic rTMS we used a novel small stimulation coil, which was specially designed to treat rats, and investigated brain tissue using in vivo localized proton magnetic resonance spectroscopy (MRS) and post mortem histological analysis. Histology was based on a modified stereology method in combination with immunohistochemistry applying antibodies against OX-6, OX-42, ED, and GFAP to detect any microglial and/or astrocytic activation 48 h after the last TMS session. Conscious rats were treated with a daily suprathreshold rTMS regimen of 1000 stimuli applied on 5 consecutive days at a frequency of 1 Hz. In comparison with control animals receiving magnetic stimulation over the lumbar spine, quantitative evaluations of cerebral metabolite concentrations by proton MRS revealed no significant alterations of N-acetyl-aspartate, creatine and phosphocreatine, choline-containing compounds, myo-inositol, glucose and lactate after chronic rTMS. Similarly to the in vivo results, post mortem histology revealed no changes in microglial and astrocytic activation after rTMS. In conclusion, these data provide support for the safety of chronic rTMS. However, they do not exclude acute changes on neurotransmitters systems or other physiologic responses during or directly after the rTMS treatment. PMID- 12765851 TI - MRI investigation of temporal lobe structures in bipolar patients. AB - Previous anatomical MRI studies have suggested abnormalities in amygdala volumes in bipolar disorder, whereas hippocampus, temporal lobe (TL), and superior temporal gyri (STG) measures have been reported to be normal. This study further investigated the existence of anatomical abnormalities in these brain structures in bipolar subjects, to attempt to replicate previously reported findings. Twenty four DSM-IV bipolar patients (mean age+/-S.D.=35+/-10 years) and 36 healthy controls (mean age+/-S.D.=37+/-10 years) were studied. 3D SPGR images were obtained with a 1.5T-GE Signa magnet (TR=25 ms, TE=5 ms, FOV=24 cm, slice thickness=1.5 mm, matrix-size=256 x 192). Volumetric measurements of TL, hippocampus, amygdala, and STG were performed blindly, with a semi-automated software. Bipolar patients had significantly larger left amygdala volumes compared with controls (mean volumes+/-S.D.=2.57+/-0.69 vs. 2.17+/-0.58 ml, respectively; ANCOVA, age, gender, ICV as covariates; F=4.42, df=1/55, P=0.04). The volumes of the other temporal lobe structures did not differ significantly between the two groups (ANCOVA, age, gender, and ICV as covariates, P>0.05). Our findings of enlarged left amygdala in bipolar patients are in agreement with prior MRI studies, suggesting that abnormalities in this brain structure may be implicated in pathophysiology of the illness. Longitudinal studies in high-risk offspring and first-episode patients will be needed to examine whether such abnormalities precede the appearance of symptoms, or whether they may appear subsequently as a result of illness course. PMID- 12765852 TI - Early age at onset as a risk factor for poor outcome of bipolar disorder. AB - The primary aim of our study was to investigate the effect of the age at onset (AAO) of Bipolar Disorder (BP) on the clinical course of the illness. We studied 320 subjects with a diagnosis of BP I or BP II who had been previously recruited for a genetic research protocol. All subjects gave their informed consent to participate in the study. Each subject was interviewed using the SCID I. The main clinical variables were compared between subjects with early (/=18 years) age at onset of BP (chi square tests and t-tests for independent samples). In addition, a logistic regression analysis was applied to the variables that were significantly related to earlier onset of BP in the exploratory analyses. We found a significantly earlier AAO in subjects with anxiety disorders (t=2.44, P=0.015) and rapid cycling course (t=3.16, P=0.002). When we compared a number of clinical characteristics between early and later onset of BP, subjects with early AAO had more frequent suicidal ideation/attempts (chi(2)=12.12, P=0.002), Axis I comorbidity (chi(2)=8.12, P=0.004), substance use disorders (chi(2)=5.45, P=0.019) and rapid cycling course (chi(2)=9.87, P=0.002). The Odds Ratios associated with these variables were: 1.407 (suicide ideation), 1.646 (Axis I comorbidity), 1.468 (substance abuse), and 2.082 (rapid cycling course). Overall, these results suggest a role of early AAO as a significant predictor of poor outcome in BP and, if replicated, they may have important clinical implications. PMID- 12765853 TI - Selective alterations of the first NREM sleep cycle in humans by a dopamine D1 receptor antagonist (NNC-687). AB - This paper details the first study of the effects of dopamine D1 receptor antagonism on the regulation of human sleep EEG (electroencephalogram). The investigational drug NNC-687 (NNC 01-0687/CEE 03-310) was administered to 20 healthy young men in doses of 5, 10, and 15 mg in a double blinded, placebo controlled, crossover design. In rats, dopamine D1 receptor antagonism can produce large increases in the amounts of both rapid eye-movement (REM) and non rapid eye-movement (NREM) sleep. In this study, drug effects were most prominent in the first NREM period. D1 antagonism markedly reduced the peak-amplitude of delta EEG waves but increased their instantaneous frequency as well as enhancing the total number, incidence, and burst-duration of sleep spindles. The length of the first NREM period was increased up to 47% over baseline. Despite these large increases in NREM sleep time, the amount of delta EEG power accumulated over the first NREM period was conserved at baseline levels. We note that the sleep-EEG profile of D1 antagonism is very similar to that of GABAA (gamma-aminobutyric acid) receptor modulators and suggest that D1 antagonism may alter the properties of the neuronal networks which generate delta and spindle, and K-complex EEG waveforms through the upstream modulation of GABAA receptor activity. PMID- 12765854 TI - Human impulsive aggression: a sleep research perspective. AB - Impulsive aggression is commonly associated with personality disorders, in particular antisocial and borderline personality disorders as well as with conduct disorder and intermittent explosive disorder. The relationship between impulsive aggression and testosterone is well established in many studies. One of the aims of this study was to characterize the relationship between earlier mentioned different categorical psychiatric diagnosis describing human impulsive aggression and sleep using polysomnography and spectral power analysis. Another aim was to study the relationship between serum testosterone and sleep in persons with severe aggressive behaviour. Subjects for the study were 16 males charged with highly violent offences and ordered for a pretrial forensic psychiatric examination. The antisocials with borderline personality disorder comorbidity had significantly more awakenings and lower sleep efficiency compared with the subjects with only antisocial personality disorder. The subjects with severe conduct disorder in childhood anamnesis had higher amount of S4 sleep and higher relative theta and delta power in this sleep stage compared with males with only mild or moderate conduct disorder. The same kind of sleep architecture was associated with intermittent explosive disorder. In subgroups with higher serum testosterone levels also the amount of S4 sleep and the relative theta and delta power in this sleep stage were increased. The study gives further support to the growing evidence of brain dysfunction predisposing to severe aggressive behaviour and strengthens the view that there are different subpopulations of individuals with antisocial personality varying in impulsiveness. The differences in impulsiveness are reflected in sleep architecture as well. PMID- 12765855 TI - Clinician recognition of anxiety disorders in depressed outpatients. AB - The recognition of anxiety disorders in depressed patients has potential clinical significance because their presence predicts poorer outcome and may influence treatment selection. In routine clinical settings, an unstructured diagnostic interview is typically used to assess patients at the initiation of treatment. Unstructured interviews, however, may result in missed diagnoses, with potential negative clinical consequences. The goals of the present study were to examine whether anxiety disorders are less frequently identified using a routine unstructured clinical evaluation than a semi-structured diagnostic interview in patients with a principal diagnosis of major depressive disorder (MDD), and to determine patients' desire for treatment for comorbid anxiety disorders. Psychiatric outpatients with MDD were evaluated with either a semi-structured or an unstructured diagnostic interview. Current DSM-IV anxiety disorder diagnoses were compared in the two, nonoverlapping, groups of depressed psychiatric outpatients seen in the same practice setting. Patients with comorbid anxiety disorders who were interviewed with the semi-structured interview were asked if they wanted treatment to address their anxiety symptoms. Individuals interviewed with the semi-structured interview were diagnosed with significantly more current anxiety disorders than individuals who were assessed with an unstructured interview. There was variability in patients' desire for treatment of the different anxiety disorders, though for each disorder the majority of patients wanted treatment to address the anxiety symptoms. In psychiatric outpatients with a principal diagnosis of MDD psychiatrists underrecognize anxiety disorder comorbidity for which patients want treatment. PMID- 12765856 TI - Patients' perception of their depressive illness. AB - Perception of illness has been described as an important predictor in the medical health psychology literature, but has been given little attention in the domain of mental disorders. The patient's Perception of Depression Questionnaire (PDIQ) is a newly developed measure whose factor structure and psychometric properties were evaluated on a sample of 174 outpatients meeting criteria for major depressive disorder. The clinical utility of the questionnaire was assessed on a sub-sample of 121 participants in a study of acupuncture treatment for depression. The questionnaire has four subscales, each with high internal consistency and high test-retest reliability. These four subscales are: Self Efficacy, which reflects perceived controllability of the illness, Externalizing, which reflects attributing the illness to external causes, Hopeless/Flawed, which reflect a belief that depression is a personal trait and therefore there is little hope for cure, and Holistic, which reflects a belief in alternative therapies. Although the PDIQ did not predict outcome, its subscales were related to adherence to treatment, treatment preference, expectations, and therapeutic alliance. The subscales have adequate convergent/discriminant validity and are clinically relevant to aspects of treatment provision. PMID- 12765857 TI - Influence of gender and hemispheric lateralization on heat pain perception in major depression. AB - Increased incidence of clinical pain complaints from patients with major depression, as well as increased experimental pain thresholds have been reported. The basis of this phenomenon remains unclear, as well as its relation to medication, clinical recovery, gender and lateralization of hemispheric function. We aimed to further elucidate heat pain perception in depression applying a testing battery including assessment (on both arms) of warmth perception, heat pain perception and heat pain tolerance, and the jaw opening reflex (duration of ES2 component) as a putative indicator of descending pain inhibition. The battery was applied to 20 patients and 20 age- and sex-matched controls. Patients were assessed: on admission (acutely depressed, off-medication), few days after admission (depressed, on medication), and after clinical recovery (mostly on medication), and controls at corresponding intervals. Significant elevated heat pain thresholds were found off and on medication in the acute stage (mainly in women) and after recovery on the right arm only. Elevated heat pain tolerance (on the right arm only) was seen in medicated patients in the acute and recovered stage. Significant prolongation of ES2 duration was only found in acutely depressed patients off medication. While confirming hypalgesia to heat pain in major depression, our findings demonstrate a close relation to gender and strong influence of lateralization after recovery. Altered pain processing at brain stem level might only partially be responsible for the observed finding. PMID- 12765858 TI - HIV-1-positive female migrants in Northrhine-Westphalia--relevant, but unfocussed problem? AB - Data of 204 HIV-1-positive female migrants from Sub-Saharan Africa, Asia and European countries other than Germany were analysed and compared to those of 282 German female virus carriers. Because not much is known about this subpopulation, the first step was to get an overview about the existing data and to describe them statistically in order to design prospective studies. A first result was that female migrants generally are tested for HIV-1 at a younger age than the control subjects. Predominant infection mode in the migrant population was heterosexual intercourse, whereas in the controls there were significantly more i.v. drug users. Distribution to the CDC-stages, CD4+-cell count and plasma viral load did not differ in both groups except for a higher percentage of AIDS-defined individuals among the controls. Surprisingly, significantly more migrants were treated with antiretroviral drugs than in the control group. However, both groups showed the same pattern of AIDS-defining diseases, only tuberculosis occurs more often in migrants. The data show that migrants are a vulnerable subpopulation which can be once being integrated in the health system of the host country effectively treated. - Results are discussed with respect to the international literature. The urgent need for adequate preventive strategies is underlined. PMID- 12765859 TI - Is anal carcinoma a HAART-related problem? AB - BACKGROUND: There is evidence that frequency of AIN/AC rises in HIV+ individuals treated with HAART whilst frequency of most other opportunistic neoplasms declines with immune reconstitution. METHOD: 1472 patients were screened 3655 times for ACA as a strong risk factor for AIN/AC. The periods 1985 to 1995 (before introduction of PI in Germany) and 1996 to 2001 were compared. In addition, 10 cases of AC are described. Time between HIV-infection and AC as well as time between ACA and AC is assessed, pre-treatment with HAART, age, CD4-cell count and CDC-stage at timepoint of diagnosis of AC is mentioned. RESULTS: There are significantly higher numbers of ACA+ screens in the years after 1996 (p<0.001) independent of number of CD4+ T-lymphocytes. AIN/AC occurs more often in HIV+ individuals, preferably in advanced disease stages, at younger age and within a shorter time after first signs of ACA than in HIV-negative population. AC occurs more likely in patients pretreated with HAART, AC tumor stage is ACIS in most cases. - CONCLUSION: Analogical to AIN/AC, frequency of ACA seems to rise under HAART (regimen including PI) although risk for opportunistic diseases usually decreases under HAART. Similarity of AC to ICC in HIV+ women is striking and AC could become part of the list of AIDS-indicator diseases. According to pre existing cost-effectiveness calculations [6], screening for AC (including physical examination, proctoscopy, cytology and biopsy of all suspect lesions) should be performed every 2-3 years in patients with CD4+ T-lymphocytes >500/microl and yearly in patients with CD4+ T-lymphocytes <500/microl. Existing ACA should be treated thoroughly. The role of serum HIV load in development and progression of ACA [20], and in consequence of AIN/AC needs further investigation. PMID- 12765860 TI - Toxoplasma gondii infection, atopy and autoimmune disease. AB - While for many allergens a dose-response relationship has clearly been established the association between early childhood exposure to cat allergens and risk of sensitisation is still controversial and even inverse relations have been described. At the same time, a negative association between Toxoplasma gondii (T. gondii) infection and atopic diseases has been found in epidemiologic studies. As cats are the major source of T. gondii oocysts, we hypothesize that cat allergen exposure might be a confounder in the association between T. gondii infection and allergic diseases. Furthermore, it is well known that atopic diseases are T helper cell 2 (Th2) driven while autoimmune diseases are dominated by a Th1 response. A counterbalance between Th1 and Th2 cells has been postulated. However, it has been shown recently that subjects with autoimmune disease are more likely to suffer from atopic diseases. Therefore, similar cytokine profiles might underlie these disorders. If this holds true it can be hypothesised that T. gondii infection is associated with a lower prevalence of autoimmune disease. PMID- 12765861 TI - Survey about the knowledge of the HIV infection amongst recruits of the German military. AB - AIMS: In the complex organizational unit of the German military (Bundeswehr), certain social conflicts and tensions may occur because of potential extreme situations and specific areas of conflict. The aim of this study was to obtain information from German military recruits regarding their knowledge of and behavioral patterns on the subject of HIV/AIDS. METHODS: 219 Soldiers in the military barracks in the state of Rhineland-Palatinate (Germany) were surveyed by means of a questionnaire (return rate 96.8%). The questionnaire comprised the following subjects: level of and requirements for information, fear of infection, behavioral pattern when in contact with HIV-infected persons as well as attitude towards governmental regulations. RESULTS: 4% of the surveyed soldiers considered their own knowledge about the subject of HIV/AIDS to be very good, 44% as good, 41% as sufficient and 7% as insufficient. 74% considered education to be necessary, and 26% felt it was not necessary. 68% of the surveyed subjects indicated that their protection of choice against infection was fidelity in partnership. 25% were of the opinion that an HIV-infected soldier should be discharged from military services and almost 20% supported that the entire barracks should be informed of such a case. 61% of those who were in favor of an obligatory registration by name (36%) also supported continued observation of the person infected with HIV. According to the surveyed soldiers, certain groups such as blood donors (86%), medical personnel (71%) as well as enlisted soldiers (28%) should receive mandatory testing. CONCLUSION: The relatively broad spectrum of opinions indicated the necessity to offer sound information and raised the question of an official regulation. PMID- 12765862 TI - The value of heparin concentration monitoring in off-pump coronary bypass surgery. AB - BACKGROUND/OBJECTIVE: In recent years, a remarcable increase in off-pump coronary bypass surgery (OPCAB) was observed. The identical anticoagulation treatment in cardiac surgery with cardiopulmonary bypass (CPB) and OPCAB makes the hemodynamic management in OPCAB procedures essential, since a perioperative compensation of blood loss is difficult and the hemodynamic stability has to be maintained by infusions. The aim of this study was to evaluate the circulating heparin concentration measurement in OPCAB patients by using the heparin/protamine titration method with Hepcon/HMSplus (HMS). PATIENTS AND METHOD: In 8 patients (3F/5M) undergoing elective OPCAB, the calculated heparin dose was administered and heparin concentration was registered together with activated clotting time (ACT) 5 min after administration. Measurements were carried out in 45 min periods and additional heparin was administered if the measured heparin concentration was lower than the calculated. The protamin dose was also automatically calculated by HMS. RESULTS: The mean operation time was 155 +/- 36 min (80-210) with 2.3 bypass grafts per patient. There was a significant difference between the demand for heparin as determined by HMS and the conventionally calculated value (p<0.05). Similarly, the calculated and measured concentration of heparin diverged significantly from each other (p<0.05), independent of ACT. In spite of an initially higher administration of heparin as calculated by HMS, the concentration of circulating heparin was below the estimated value in 7 patients which made a further administration of heparin necessary. The 24 h postoperative blood loss was 550 +/- 176 mL (300-850). No rethoracotomy for bleeding was necessary. CONCLUSIONS: HMS could be a useful method for appropriate anticoagulative treatment in OPCAB procedures, if there is a sufficient hemodynamic management with restrictive administration of infusions guaranted during surgery. PMID- 12765863 TI - Gene expression of type I collagen in neoplastic chondrocytes. AB - In the present examination, we detected type I collagen mRNA in neoplastic chondrocytes in osteochondromas, typical benign bone neoplasms. We believe that the cells involved in >chondroid bone< appearing in osteochondromas temporally express cartilage phenotypes and then change directly into bone-forming cells that survive in the >chondroid bone< until the tissue is resorbed and remodelled into true bone tissue. PMID- 12765865 TI - The 1971 BOHS Hygiene Standard for Wide-band Noise. PMID- 12765866 TI - Chronic fatigue and organophosphate pesticides in sheep farming: a retrospective study amongst people reporting to a UK pharmacovigilance scheme. AB - The Department of Health has recently published a report from the CFS/ME Working Group which concluded that chronic fatigue syndrome (CFS) should be recognized as a chronic illness. Symptoms consistent with CFS are often reported by people who consider their health has been affected by exposure to pesticides, but the Working Group concluded that this type of exposure is not a common trigger for the syndrome. The Veterinary Medicines Directorate (VMD) collects self-assessed reports of ill health in humans associated with veterinary medicines under their Suspected Adverse Reaction Surveillance Scheme. The reporters have mainly been sheep farmers. These reports were used to investigate the possible relationship between chronic fatigue (CF) and exposure to organophosphate pesticides in sheep farming. The overall aim of the study was to investigate a possible association between exposure to organophosphates and the development of CF amongst people who consider their health has been affected by pesticides in sheep farming. The hypothesis investigated was that repeated exposure to organophosphate pesticides in sheep dip may increase the probability of developing CF. A group of mostly sheep farmers who had reported to the VMD surveillance scheme were identified. We planned to use a retrospective case-control study design but the initial symptoms reports were not sufficiently reliable to enable this. The study population was asked to complete two questionnaires. The first questionnaire was designed to identify the history of exposure of subjects to organophosphate pesticides, and their exposure was then reconstructed using a metric specifically developed for this purpose. The second questionnaire collected detailed information to identify whether the subjects had CF when they originally reported to the VMD and at the time of the survey. The questionnaire was sent to a total of 206 subjects, of whom 28 had moved home. A total of 37% of the remaining 178 subjects participated. There was a high prevalence of CF amongst those who completed the questionnaire and this has generally persisted since the subjects reported to the VMD. Higher CF scores were associated with higher exposure to organophosphate pesticides. CF is very common amongst those who consider their health was affected by pesticides and we have shown there is limited evidence of an association between exposure to organophosphates and CF. Further research is needed to investigate the cause of this syndrome amongst farmers exposed to pesticides. PMID- 12765867 TI - The performance of laboratories analysing alpha-quartz in the Workplace Analysis Scheme for Proficiency (WASP). AB - The Workplace Analysis Scheme for Proficiency (WASP) is a proficiency testing (PT) scheme for the analysis of occupational hygiene and environmental air samples and is operated in the United Kingdom by the Health and Safety Laboratory (HSL) on behalf of the Health & Safety Executive (HSE). One of the 26 analytes available to laboratories is silica (alpha-quartz) on 25 mm Gelman GLA5000 filters. This paper investigates the performance of laboratories participating in the scheme since the HSL took over the production of the samples in 1998. The average relative standard deviation (RSD) of results obtained by a laboratory is 11.5%. This is reduced to 8.5% when the values from laboratories using indirect analytical methods are excluded. Laboratories using indirect analytical methods accounted for some of the most variable data. For the on-filter analytical methods the data suggested a relationship between relative standard deviation and loading that increased gradually from +/-4% at high analyte levels to +/-10-15% at low levels. The average precision estimate for the on-filter analytical methods was found to be 5.6% RSD for the infrared technique and 6.7% RSD for the X-ray diffraction technique. These figures compare favourably with those reported in the published HSE methods. No significant difference was found between the average result reported by laboratories using on-filter infrared (IR) analysis and the average result reported by laboratories using on-filter X-ray diffraction (XRD) analysis. An ANOVA analysis found the repeatability estimate was just as large as the 'between laboratory' variation for both the XRD and IR on-filter analysis techniques. When a limited number of 'realistic' samples were included in the scheme, XRD analysis was found to perform slightly better than IR analysis. The performance of laboratories in the WASP scheme compares very favourably with other published data from a PT scheme where indirect silica analytical methods are predominately used. PMID- 12765868 TI - Overall mass transfer coefficient for pollutant emissions from small water pools under simulated indoor environmental conditions. AB - Small chamber tests were conducted to experimentally determine the overall mass transfer coefficient for pollutant emissions from still aqueous solutions under simulated indoor (residential or occupational) environmental conditions. The tests covered six organic compounds with a Henry's constant range from 3.33 x 10( 7) to 3.67 x 10(-3) (atm m(3)/mol). The estimated overall liquid phase mass transfer coefficients for still solutions varied from 1.8 x 10(-6) to 5.7 x 10( 3) m/h; the estimated liquid phase mass transfer coefficients were 9.7 x 10(-3) m/h for the reference compound (oxygen) and 5.00 x 10(-3) to 6.04 x 10-(3) m/h for the test compounds. An empirical model is proposed to estimate the overall mass transfer coefficient, which can be used to predict pollutant emissions from still aqueous solutions (e.g. pools and puddles) in indoor environments. PMID- 12765869 TI - Respiratory symptoms and occupational exposures in New Zealand plywood mill workers. AB - OBJECTIVES: To study work exposure and respiratory symptoms in New Zealand plywood mill workers. METHODS: Personal inhalable dust (n = 57), bacterial endotoxin (n = 20), abietic acid (n = 20), terpene (n = 20) and formaldehyde (n = 22) measurements were taken and a respiratory health questionnaire was administered to 112 plywood mill workers. RESULTS: Twenty-six percent of the dust exposures exceeded 1 mg/m(3), however, none of the samples exceeded the legal limit of 5 mg/m(3) [geometric mean (GM) = 0.7 mg/m(3), geometric standard deviation (GSD) = 1.9]. Workers in the composer area (where broken sheets are joined together) were significantly (P < 0.01) more highly exposed. Endotoxin levels were low to moderate (GM = 23.0 EU/m(3), GSD = 2.8). Abietic acid levels ranged from 0.3 to 2.4 micro g/m(3) (GM = 0.7 micro g/m(3), GSD = 1.8) and were significantly (P < 0.05) higher for workers in the composer area of the process. Geometric mean levels of alpha-pinene, beta-pinene and Delta(3)-carene were 1.0 (GSD = 2.7), 1.5 (GSD = 2.8) and 0.1 (GSD = 1.4), respectively, and alpha-pinene and beta-pinene levels were significantly (P < 0.001) higher for workers in the 'green end' of the process, up to and including the veneer dryers. Formaldehyde levels ranged from 0.01 to 0.74 mg/m(3) [GM = 0.08 mg/m(3) (= 0.06 p.p.m.), GSD = 3.0]. Asthma symptoms were more common in plywood mill workers (20.5%, n = 112) than in the general population [12.8%, n = 415, adjusted OR (95% CI) = 1.5 (0.9 2.8)]. Asthma symptoms were associated with duration of employment and were reported to lessen or disappear during holidays. No clear association with any of the measured exposures was found, with the exception of formaldehyde, where workers with high exposure reported more asthma symptoms (36.4%) than low exposed workers [7.9%, adjusted OR (95% CI) = 4.3 (0.7-27.7)]. CONCLUSIONS: Plywood mill workers are exposed to inhalable dust, bacterial endotoxin, abietic acid, terpenes and formaldehyde, and they appear to have an increased risk of developing work-related respiratory symptoms. These symptoms may be due to formaldehyde exposure, although a potential causal role for other exposures cannot be excluded. PMID- 12765870 TI - Sensory irritation due to methyl-2-cyanoacrylate, ethyl-2-cyanoacrylate, isopropyl-2-cyanoacrylate and 2-methoxyethyl-2-cyanoacrylate in mice. AB - The expiratory bradypnoea indicative of upper airway irritation in mice was evaluated during a period of 60 min of nasal exposure to methyl-2-cyanoacrylate, ethyl-2-cyanoacrylate, isopropyl-2-cyanoacrylate and 2-methoxyethyl-2 cyanoacrylate vapors using nose only exposure. Irritation of the upper respiratory tract caused a concentration-dependent decrease in the respiratory rate. The maximum effect occurred within the first 10 min of exposure and was followed by a drop-off in the response during the remainder of the exposure period. The airborne concentration resulting in a 50% decrease in the respiratory rate of mice (RD(50)) was calculated for each chemical. The results show that the four chemicals had similar irritant potencies. The RD(50) values of methyl-2 cyanoacrylate, ethyl-2-cyanoacrylate, isopropyl-2-cyanoacrylate and 2 methoxyethyl-2-cyanoacrylate were 1.4, 0.7, 0.6 and 1.0 p.p.m. Tentative estimates of threshold limit values showed that 0.1 RD(50) was closer to the values recommended by the American Conference of Governmental Industrial Hygienists for methyl- and ethyl-2-cyanoacrylate than 0.03 RD(50). On the basis of a threshold limit value for short-term exposure limit (TLV STEL) equal to 0.1 RD(50), the TLV STELs for the four cyanoacrylates should not exceed 0.1 or 0.2 p.p.m. PMID- 12765871 TI - Permeation of 70% isopropyl alcohol through surgical gloves: comparison of the standard methods ASTM F739 and EN 374. AB - Standard test methods ASTM F739 and EN 374 were compared by assessing the permeation of 70% isopropyl alcohol (2-propanol) through seven brands of surgical gloves. The two standards differ in the flow rates of the collection medium and in the chemical permeation rate at which the breakthrough time (BTT) is detected, the EN detection level being 10 times higher than the permeation rate used by ASTM. In a departure from the EN standard method, a 4 h testing time was used instead of 8 h. All of the tested gloves were from the same manufacturer and were made from either natural rubber (NR) (six brands) or chloroprene rubber (CR) (one brand). Two of the NR glove brands were double layered. For the thin NR gloves (0.22, 0.28 and 0.27 mm) the permeation rates were higher throughout the tests with a flow rate of 474 ml/min (EN) of the collection medium (nitrogen) compared with the permeation rates obtained with a flow rate of 52 ml/min (ASTM). These resulted in BTTs of 4.6, 6.5 and 7.6 min (EN) and 4.8, 6.5 and 9.1 min (ASTM), respectively. No statistical difference could be observed between the BTT values obtained with the two standard methods for any of the thin gloves. Thus, although the ASTM standard has a lower criterion for the detection of permeation, it does not necessarily produce shorter BTTs. For the better barriers the methods yielded more equivalent permeation rate curves and thus the EN BTTs were longer than the ASTM BTTs: the EN results were 21, 80, 122 and >240 min compared with the ASTM results of 12, 32, 38 and 103 min for glove thicknesses of 0.37 (NR), 0.22 + 0.22 (double layered NR), 0.31 + 0.29 (double layered NR) and 0.19 mm (CR), respectively. PMID- 12765872 TI - The permeability of surgical gloves to seven chemicals commonly used in hospitals. AB - Disinfectants may cause adverse effects directly on the skin or systemically by permeating through the skin. In this study breakthrough times were measured for surgical gloves with chemicals which are commonly used in healthcare. Classical methods of analytical chemistry were tailored for the permeation tests, which were carried out according to the European standard EN 374 and the American standard ASTM F739. An exception to the EN 374 standard was made by using a 4 h testing time instead of 8 h. The gloves did not exhibit permeation of potassium hydroxide (45%), sodium hypochlorite (13%) or hydrogen peroxide (30%). Furthermore, neither glutaraldehyde (2%) nor chlorhexidine digluconate (4%) in the commercial disinfectant solutions studied exhibited permeation. Slight permeation of peracetic acid (0.35%) and acetic acid (4%) from a disinfectant agent was observed through single layered natural rubber materials. Clear evidence of formaldehyde permeation was detected through single layered natural rubber gloves, where the ASTM breakthrough times were 17-67 min, but the permeation rates were not high enough for breakthrough to have occurred according to the EN standard. The gloves in this study which offered most protection from chemical permeation were the chloroprene gloves and the thick double layered natural rubber gloves. PMID- 12765873 TI - Exposure to brake dust and malignant mesothelioma: a study of 10 cases with mineral fiber analyses. AB - OBJECTIVES: A large number of workers in the USA are exposed to chrysotile asbestos through brake repair, yet only a few cases of malignant mesothelioma (MM) have been described in this population. Epidemiologic and industrial hygiene studies have failed to demonstrate an increased risk of MM in brake workers. We present our experience of MM in individuals whose only known asbestos exposure was to brake dust and correlate these findings with lung asbestos fiber burdens. METHODS: Consultation files of one of the authors were reviewed for cases of MM in which brake dust was the only known asbestos exposure. Lung fiber analyses were performed using scanning electron microscopy (SEM) in all cases for which formalin-fixed or paraffin-embedded lung tissue was available. RESULTS: Ten cases of MM in brake dust-exposed individuals were males aged 51-73 yr. Nine cases arose in the pleura and one in the peritoneum. Although the median lung asbestos body count (19 AB/g) is at our upper limit of normal (range 0-20 AB/g), half of the cases had levels within our normal range. In every case with elevated asbestos fiber levels by SEM, excess commercial amphibole fibers were also detected. Elevated levels of chrysotile and non-commercial amphibole fibers were detected only in cases that also had increased commercial amphibole fibers. CONCLUSIONS: Brake dust contains exceedingly low levels of respirable chrysotile, much of which consists of short fibers subject to rapid pulmonary clearance. Elevated lung levels of commercial amphiboles in some brake workers suggest that unrecognized exposure to these fibers plays a critical role in the development of MM. PMID- 12765874 TI - Biological monitoring of low level occupational xylene exposure and the role of recent exposure. AB - The correlation between low level time-weighted average (TWA) atmospheric xylene exposure (p.p.m.) and urinary methylhippuric acid (MHA) expressed per gram of creatinine was examined. Subjects were recruited from workplaces that utilized xylene. Ambient monitoring of o-, m- and p-xylene isomers was carried out using passive diffusion vapour monitors. Adjusted (post-shift minus pre-shift) and post shift urinary levels of xylene metabolites (2-, 3- and 4-MHA) were determined by GC-MS. Twenty subjects were recruited into the study. Total xylene TWA exposures were 3.36 +/- 3.63 p.p.m. (mean +/- SD) with a range of 0.03-14.44 p.p.m. The r(2) values for the regression equations between xylene exposure and individual and total adjusted MHA isomers were 0.390, 0.709, 0.677 and 0.631 for o-, m-, p- and total xylenes, respectively, which was greater than the respective correlations between non-adjusted samples. In conclusion, biological monitoring of occupational xylene exposure at levels <15 p.p.m. using urinary MHA showed a good correlation with atmospheric levels and is a valid complement to ambient monitoring. Even though occupational xylene exposure in the workplaces studied was generally low, MHA was found in the pre-shift urine of all workers and the use of adjusted values showed modest improvements in correlations. Recent exposure prior to sampling, either from occupational or non-occupational sources, should be considered when biological monitoring of xylene is undertaken. Extrapolation of data from this study predicted a MHA concentration in post-shift urine of 1.3 g/g creatinine after exposure to a TWA of 100 p.p.m. xylene. PMID- 12765875 TI - The comfort and effectiveness of hearing protection devices. PMID- 12765878 TI - Phenazine-1-carboxylic acid, a secondary metabolite of Pseudomonas aeruginosa, alters expression of immunomodulatory proteins by human airway epithelial cells. AB - Pseudomonas aeruginosa is a gram-negative bacterium that causes both acute and chronic lung disease in susceptible patient populations. P. aeruginosa secretes numerous proteins and secondary metabolites, many of which have biological effects that likely contribute to disease pathogenesis. An unidentified small molecular-weight factor was previously reported to increase IL-8 release both in vitro and in vivo. To identify this factor, we subjected the <3-kDa fraction from P. aeruginosa-conditioned medium to HPLC analysis. A peak fraction that stimulated IL-8 release was found by mass spectrometry to have a molecular mass (MM) of 224 Da. On the basis of this MM and other biochemical properties, we hypothesized that the factor was phenazine-1-carboxylic acid (PCA). Subsequent studies and comparison with purified PCA confirmed this hypothesis. Purified PCA exhibited a number of biological effects in human airway epithelial cells, including increasing IL-8 release and ICAM-1 expression, as well as decreasing RANTES and monocyte chemoattractant protein-1 (MCP-1) release. PCA also increased intracellular oxidant formation as measured by electron paramagnetic resonance and by an intracellular oxidant-sensitive probe. Antioxidants inhibited PCA dependent increases in IL-8 and ICAM-1, suggesting that oxidants contributed to these effects. However, in contrast to the related phenazine compound pyocyanin, PCA did not oxidize NAD(P)H at physiologically relevant pH, providing preliminary evidence that PCA and pyocyanin may have distinct redox chemistries within the cell. Thus PCA is a biologically active factor secreted by P. aeruginosa that has several activities that could alter the host immune and inflammatory response and thereby contribute to bacterial disease pathogenesis. PMID- 12765879 TI - Inhaled nitric oxide increases surfactant protein gene expression in the intact lamb. AB - Inhaled nitric oxide (iNO) is used to treat a number of disease processes. Although in vitro data suggest that nitric oxide (NO) alters surfactant protein gene expression, the effects in vivo have not been studied. The objective of this study was to evaluate the effects of iNO on surfactant protein (SP)-A, -B, and -C gene expression in the intact lamb. Thirteen 4-wk-old lambs were mechanically ventilated with 21% oxygen and received iNO at 40 ppm (n = 7) or vehicle gas (n = 6) for 24 h. Peripheral lung biopsies were obtained at 0, 12, and 24 h and analyzed for surfactant mRNA, protein, and total DNA content. Inhaled NO increased SP-A and SP-B mRNA content by 80% from 0 to 12 h and by 78 and 71%, respectively, from 0 to 24 h. There was an increase in SP-A and SP-B protein content by 45% from 0 to 12 h, and a decrease by 70 and 65%, respectively, from 0 to 24 h. DNA content was unchanged. The mechanisms and physiological effects of these findings warrant further investigation. PMID- 12765880 TI - Pulmonary PKG-1 is upregulated following chronic hypoxia. AB - Recent studies from our laboratory indicate that pulmonary vasodilatory responses to exogenous nitric oxide (NO) are attenuated following chronic hypoxia (CH) and that this NO-dependent vasodilation is mediated by cGMP. Similarly, we have demonstrated that CH attenuates vasodilatory responses to the cGMP analog 8 bromoguanosine 3',5'-cyclic monophosphate (8-BrcGMP). We hypothesized that attenuated pulmonary vasodilation to 8-BrcGMP following CH is mediated by decreased protein kinase G-1 (PKG-1) expression/activity. Therefore, we examined vasodilatory responses to 8-BrcGMP (1 microM) in isolated, saline-perfused lungs from control and CH (4 wk at barometric pressure of 380 mmHg) rats in the presence of the competitive PKG inhibitor Rp-beta-phenyl-1, N2-etheno-8 bromoguanosine 3',5'-cyclic monophosphorothionate (30 microM) or the highly specific PKG inhibitor KT-5823 (10 microM). PKG-1 expression and activity were determined in whole lung homogenates from each group, and vascular PKG-1 levels were assessed by quantitative immunohistochemistry. PKG inhibition with either Rp 8-Br-PET-cGMPS or KT-5823 diminished vasodilatory responses to 8-BrcGMP in lungs from both control and CH rats, thus indicating a role for PKG in mediating reactivity to 8-BrcGMP in each group. However, in contrast to our hypothesis, PKG 1 levels were approximately twofold greater in lungs from CH rats vs. controls, and furthermore, this upregulation was localized to the vasculature. This correlates with an increase in PKG activity following CH. We conclude that PKG-1 is involved in 8-BrcGMP-mediated vasodilation; however, attenuated pulmonary vasodilation following CH is not associated with decreased expression/activity of PKG-1. PMID- 12765882 TI - Anaesthesia and SARS. PMID- 12765881 TI - Zinc chelators inhibit eotaxin, RANTES, and MCP-1 production in stimulated human airway epithelium and fibroblasts. AB - Asthma is characterized by an increased production of eosinophil-active C-C chemokines by the airway epithelium. Recent studies have identified the presence of important quantities of labile zinc in the conducting airways. We hypothesized that modulation of this labile zinc could influence the production of proinflammatory chemokines in respiratory epithelial cells. The zinc chelator N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine (TPEN) and the heavy metal chelator 2,3-dimercapto-1-propanesulfonic acid (DMPS) were used to reduce the labile zinc content of A549, BEAS-2B, and HFL-1 cells. Northern blot analysis and RNase protection assay were used to study the effects of the zinc chelators on mRNA expression. DMPS and TPEN specifically inhibited the production of eotaxin, regulated on activation, normal T-cell expressed, and presumably secreted, and monocyte chemotactic protein-1 in TNF-alpha-stimulated respiratory epithelial cells and fibroblasts through labile zinc chelation. The inhibitory effects of DMPS and TPEN were associated with the decreased binding of the zinc-finger transcription factor GATA-1, whereas no change in NF-kappaB activation was observed. Together these results demonstrate that modulation of the labile pool of zinc can regulate gene expression and protein synthesis of C-C chemokines in lung epithelial cells and fibroblasts. PMID- 12765883 TI - When should we transfuse critically ill and perioperative patients with known coronary artery disease? PMID- 12765884 TI - Old dog--new (ma)trix. PMID- 12765885 TI - Do genes influence outcome from anaesthesia? PMID- 12765886 TI - Effects of dexamethasone on clinical course, C-reactive protein, S100B protein and von Willebrand factor antigen after paediatric cardiac surgery. AB - BACKGROUND: Anti-inflammatory treatment with glucocorticoids during cardiopulmonary bypass can reduce inflammatory mediator release, but the effects of glucocorticoid on outcome are controversial. METHODS: We studied the effects of dexamethasone on clinical course, C-reactive protein, von Willebrand factor antigen (vWf:Ag) and S100B in a randomized masked study of children after open cardiac surgery. Twenty children weighing >10 kg received dexamethasone (1 mg kg( 1)) and 20 controls received saline after induction of anaesthesia. We measured vWf:Ag as a marker of endothelial activation, S100B as a marker of cerebral protein release and C-reactive protein as a marker of inflammatory activity. Oxygenation, body temperature, fluid balance, leucocyte and platelet counts, days in the intensive care unit (ICU) and days on mechanical ventilation were noted. RESULTS: Dexamethasone decreased C-reactive protein concentration on the first postoperative day (P<0.05), but did not affect the release of vWf:Ag or S100B. There was no significant difference in oxygenation, body temperature, fluid balance, leucocyte and platelet counts, days in the ICU or days on mechanical ventilation between the placebo and dexamethasone-treated groups. CONCLUSION: Administration of dexamethasone before cardiopulmonary bypass for paediatric cardiac surgery decreased the inflammatory response, but did not affect the immediate features after surgery or changes in vWf:Ag or S100B. PMID- 12765887 TI - Left ventricular regional wall motion abnormalities during pneumoperitoneum in children. AB - BACKGROUND: In adult patients, certain levels of PEEP (16 and 20 cm H(2)O) have been associated with left ventricular (LV) regional wall motion abnormalities. Since any increase in intra-abdominal pressure (IAP) exerted by a pneumoperitoneum is transmitted to the intrathoracic cavity, similar effects on LV regional wall motion cannot be ruled out. METHODS: To investigate the effects of pneumoperitoneum on LV regional wall motion, we performed a post hoc analysis of a transoesophageal echocardiography study in eight small children (mean age 3 yr, range 15-63 months) undergoing laparoscopic herniorrhaphy under anaesthesia with sevoflurane in nitrous oxide/oxygen and a PEEP of 5 cm H(2)O. During carbon dioxide insufflation, end-tidal carbon dioxide concentration ()was kept constant by increasing minute volume. RESULTS: An IAP of 12 mm Hg caused significant septal hypokinesia compared with baseline, while anterior and posterior wall motion was not affected. In addition, a lateral hyperkinesia occurred, though this change was not statistically significant. CONCLUSIONS: Pneumoperitoneum may affect LV regional wall motion in paediatric patients undergoing laparoscopic surgery. PMID- 12765889 TI - Sedation caused by clonidine in patients with spinal cord injury. AB - BACKGROUND: In patients with spinal cord injury, cephalad spread of intrathecal (i.t.) medication could be delayed. METHODS: We used bispectral index and an observer scale to assess sedation after two different doses of i.t. clonidine in patients with or without spinal cord injury. Twelve patients with neurological deficit caused by trauma (Spinal Cord Injury, SCI) were compared with patients without neurological disease. They received 10 mg of i.t. bupivacaine with clonidine, with either 50 microg (low dose, n=6) or 150 microg (high dose, n=6) at L(2)-L(3). A further 12 patients, six with spinal trauma lesion and six healthy, received i.t. bupivacaine and 150 micro g of i.m. clonidine. RESULTS: Sedation and a decrease in BIS occurred only in patients receiving 150 microg of clonidine. Onset of sedation and the decrease in BIS was delayed in most spinal cord injured patients whatever the route of administration (P<0.001). Duration of sedation was not different between the groups. Delayed sedation and decrease of BIS after i.t. clonidine in patients with spinal cord injury are similar than those observed after i.m. clonidine. CONCLUSION: A systemic effect is likely to be the main reason for sedation. PMID- 12765888 TI - Closed-loop control of propofol anaesthesia using bispectral index: performance assessment in patients receiving computer-controlled propofol and manually controlled remifentanil infusions for minor surgery. AB - BACKGROUND: In a previous study we used the bispectral index (BIS) for automatic control of propofol anaesthesia, using a proportional-integral-differential control algorithm. As control was less than optimal in some patients, we revised the constants of the control algorithm. The aim of the current study was to measure the performance of the revised system in patients undergoing minor surgery under propofol and remifentanil anaesthesia. METHODS: Twenty adult patients scheduled for body surface surgery were enrolled. Anaesthesia was manually induced with target-controlled infusions (TCI) of propofol and remifentanil. After the start of surgery, when anaesthesia was clinically adequate, automatic control of the propofol TCI was commenced using the revised closed-loop system. For patients 11-20, effect-site steering was also incorporated into the closed-loop control algorithm. Adequacy of anaesthesia during closed-loop control was assessed clinically, and by calculating the median performance error (MDPE), the median absolute performance error (MDAPE) and the mean offset of the control variable. RESULTS: The system provided adequate operating conditions and stable cardiovascular values in all patients during closed-loop control. The mean MDPE and MDAPE were -0.42% and 5.63%, respectively. Mean offset of the BIS from setpoint was -0.2. No patients reported awareness or recall of intraoperative events. CONCLUSIONS: The system was able to provide clinically adequate anaesthesia in all patients, with better accuracy of control than in the previous study. There was a tendency for more accurate control in those patients in whom the control algorithm incorporated effect-site steering. PMID- 12765890 TI - Analgesic effects of parecoxib following total abdominal hysterectomy. AB - BACKGROUND: Forty-eight ASA I-II patients undergoing total abdominal hysterectomy (TAH) were studied in a double blind, randomized placebo controlled trial of parecoxib for postoperative analgesia. METHODS: All patients were given propofol 2-4 mg kg(-1) i.v., a non-depolarizing muscle relaxant, morphine 10 mg i.v. and prochlorperazine 12.5 mg i.m. intraoperatively. Their lungs were ventilated with nitrous oxide and isoflurane 1-1.5% in oxygen. Morphine was self-administered for postoperative analgesia via a patient controlled analgesia (PCA) device. Patients were allocated randomly to receive either parecoxib 40 mg i.v. or normal saline on induction of anaesthesia. RESULTS: Twelve patients did not complete the study. Of the remaining 36 patients, there was no significant difference between the treatment groups in age, weight, ASA status, duration of surgery, or intraoperative dose of morphine. However, mean (95% CI) 24 h morphine consumption of 54 (42-65) mg in the parecoxib group was significantly (P=0.04) lower than that of 72 (58-86) mg in the placebo group. Pain intensity scores on sitting up were significantly lower (P=0.02) in the parecoxib group compared with placebo. There was no significant difference between the treatment groups in pain intensity scores at rest and on deep inspiration, or in nausea, total number of vomiting episodes, median number of rescue antiemetic doses, and sedation scores. CONCLUSIONS: Parecoxib 40 mg i.v. may be recommended in patients having TAH as it provides morphine-sparing analgesia. PMID- 12765892 TI - Effects of halogenated anaesthetics on diaphragmatic actin-myosin cross-bridge kinetics. AB - BACKGROUND: The effects of halogenated anaesthetics on cross-bridge (CB) kinetics are unclear. As halogenated anaesthetics do not markedly modify the intracellular calcium transient in the diaphragm, we used an isolated rat diaphragm preparation to assess the effects of halothane and isoflurane on CB kinetics. METHODS: The effects of halothane and isoflurane (1 and 2 minimum alveolar concentration (MAC)) on rat diaphragm muscle strips were studied in vitro (Krebs-Henseleit solution, 29 degrees C, oxygen 95%/carbon dioxide 5%) in tetanus mode (50 Hz). From the force-velocity curve and using A. F. Huxley's equations, we determined the main mechanical and energetic variables and calculated CB kinetics. RESULTS: At 1 and 2 MAC, isoflurane and halothane induced no significant inotropic effects. Whatever the concentrations tested, halothane and isoflurane did not significantly modify the CB number, the elementary force per CB, the attachment and detachment constants, the duration of the CB cycle and mean CB velocity. CONCLUSION: In the rat diaphragm at therapeutic concentrations, halogenated anaesthetics do not significantly modify CB mechanical and kinetic properties. PMID- 12765891 TI - Cerebral and lung kinetics of morphine in conscious sheep after short intravenous infusions. AB - BACKGROUND: The analgesic effects of morphine are delayed relative to its concentration in blood. The rate of equilibration of morphine between blood and brain may contribute to this delay, but the kinetics of this process have not been modelled. This was determined in conscious instrumented sheep. The lung kinetics of morphine were also determined given their importance in defining systemic kinetics after i.v. bolus administration. METHODS: Sheep were given short i.v. infusions of morphine (30 mg over 4 min). Cerebral kinetics were inferred from arterio-sagittal sinus concentration gradients and cerebral blood flow, and lung kinetics from the pulmonary artery-aortic gradient and cardiac output. These data were fitted to flow- and membrane-limited models of the kinetics in each organ. RESULTS: Morphine had minimal cardiovascular effects, did not alter cerebral blood flow and caused insignificant respiratory depression. Lung kinetics were best described by a single distribution volume (2036 ml) with a first-order loss (1370 ml min(-1)), which was attributed to deep distribution. The cerebral kinetics of morphine were characterized by a significant permeability barrier. Permeability across the barrier (7.44 ml min(-1)) was estimated with good precision, and was approximately one-fifth of the nominal cerebral blood flow. The distribution volume of morphine in the brain was estimated with less precision, but was described by a brain:blood partition coefficient of approximately 1.4. The time required for 50% equilibration between brain and blood concentrations was approximately 10.3 min. CONCLUSION: The cerebral equilibration of morphine was relatively slow, and was characterized by significant membrane limitation. PMID- 12765893 TI - Effects of sevoflurane on sympathetic neurotransmission in human omental arteries and veins. AB - BACKGROUND: Sevoflurane reduces blood pressure, the regulation of which requires an intact sympathetic neurotransmission. This study was designed to evaluate the effect of sevoflurane on the coupling between peripheral sympathetic neurones and vascular smooth muscle in isolated human omental vessels. METHODS: Segments of arteries and veins were exposed to sevoflurane 1%, 2% and 4% (corresponding to approximately 0.5, 1 and 2 MAC in humans, respectively). The vessels were studied in vitro to determine the effects on (i) isometric contraction during electrical field stimulation (EFS) or in the presence of exogenous norepinephrine (NE); (ii) electrical field stimulated release of [(3)H]-NE from vessel segments previously incubated with [(3)H]-NE; (iii) uptake of [(3)H]-NE. RESULTS: In artery segments, sevoflurane 4% attenuated the contraction induced by both EFS and exogenous NE. In vein segments, sevoflurane 4% attenuated only the EFS-induced contractions. Sevoflurane 1% and 2% had no effect. The release of [(3)H]-NE was inhibited by sevoflurane 2% and 4% in arteries and by sevoflurane 1%, 2% and 4% in veins. Sevoflurane had no effect on the uptake of [(3)H]-NE in either vessel. CONCLUSIONS: Sevoflurane depresses sympathetic neuromuscular transmission in human omental vessels by reducing neuronal NE release and NE sensitivity in arteries and by reducing NE release in veins. This could contribute to the hypotension seen during sevoflurane anaesthesia, at least at concentrations above 1 MAC. PMID- 12765894 TI - Cerebral oxygen vasoreactivity and cerebral tissue oxygen reactivity. AB - There has long been an appreciation that cerebral blood flow is modulated to ensure adequate cerebral oxygen delivery in the face of systemic hypoxaemia. There is increasing appreciation of the modulatory role of hyperoxia in the cerebral circulation and a consideration of the effects of such modulation on the maintenance of cerebral tissue oxygen concentration. These newer findings are particularly important in view of the fact that cerebrovascular and tissue oxygen responses to hyperoxia may change in disease. Such alterations provide important insights into pathophysiological mechanisms and may provide novel targets for therapy. However, before the modulatory effects of hyperoxia can be used for diagnosis, to predict prognosis or to direct therapy, a more detailed analysis and understanding of the physiological concepts behind this modulation are required, as are the limitations of the measurement tools used to define the modulation. This overview summarizes the available information in this area and suggests some avenues for further research. PMID- 12765895 TI - Sub-Tenon's administration of local anaesthetic: a review of the technique. PMID- 12765896 TI - A simple method to pass a pulmonary artery flotation catheter rapidly into the pulmonary artery in anaesthetized patients. AB - BACKGROUND: In some patients passage of a pulmonary artery flotation catheter (PAFC) into the pulmonary artery may be difficult and time consuming and the prolonged manipulation can cause ventricular arrhythmias. A simple clinical method used during general anaesthesia is presented to allow rapid passage of a PAFC into the pulmonary artery. METHODS: The operating table is positioned head up and slightly right side down to position the pulmonary valve at the highest level possible. When the balloon catheter is in the right ventricular outflow tract (indicated by premature ventricular contractions) the ventilator is paused in inspiration and the balloon catheter simultaneously passed into the pulmonary artery. RESULTS: The manoeuvre shortens the time necessary to pass the catheter into the pulmonary artery and may reduce ventricular arrhythmias. Over 5 yr, 105 PAFCs were inserted with this method without major complications. CONCLUSION: This method may reduce the risk of ventricular arrhythmias, and could be particularly useful in high-risk critically ill patients. PMID- 12765897 TI - Hyperlordosis as a possible factor in the development of spinal cord infarction. AB - A patient developed persistent symptoms and signs suggestive of partial spinal cord infarction after an operation involving the use of the hyperlordotic position. This position involves extension at the waist, such that both the head and feet are below the level of the waist. It is employed to increase surgical access to the abdomen. Where this position is adopted for a prolonged surgical procedure, existing risk factors for spinal cord ischaemia should urge caution in the use of epidural analgesia. PMID- 12765898 TI - Acute heart failure during spinal surgery in a boy with Duchenne muscular dystrophy. AB - Patients with Duchenne muscular dystrophy (DMD) are at high risk of perioperative complications. DMD may be accompanied by heart failure resulting from dystrophic involvement of the myocardium, which can be subclinical in the early stages of the disease. This case demonstrates that a normal preoperative ECG and echocardiograph cannot exclude the development of heart failure during anaesthesia in DMD patients undergoing major surgery. PMID- 12765899 TI - Unusual presentation of long QT syndrome. AB - We describe the case of a 9-yr-old child with undiagnosed long QT syndrome who experienced an intraoperative cardiac arrest after accidental intravascular injection of bupivacaine with epinephrine via a misplaced epidural catheter. PMID- 12765900 TI - Outreach critical care. PMID- 12765901 TI - The effect of haemodilution on coagulation. PMID- 12765902 TI - Unexplained fitting in patients with post-dural puncture headache. Risk of iatrogenic pneumocephalus with air rationalizes use of loss of resistance to saline. PMID- 12765903 TI - Atracurium is associated with postoperative residual curarization. PMID- 12765904 TI - Repeated intranasal capsaicin applications to treat chronic migraine. PMID- 12765905 TI - The one child family policy. PMID- 12765906 TI - Cycle helmets. PMID- 12765907 TI - Health effects of family size: cross sectional survey in Chinese adolescents. AB - AIMS: To determine whether only children differ in terms of morbidity, nutritional status, risk behaviours, and utilisation of health services from children with siblings, in China. METHODS: A cross sectional survey was carried out using self completion questionnaires, anthropometry, and haemoglobin measurement in middle schools (predominant age 12-16 years) in three distinct socioeconomic areas of Zhejiang province, eastern China. RESULTS: Data were obtained for 4197 participants. No significant differences were found between only children and those with siblings for some key indicators: underweight 19% v 18%, suicide ideation 14% v 14%, and ever smoking 17% v 15%. Only children were more likely to be overweight (4.8% v 1.5%), and to have attended a doctor (71% v 63%) or dentist (17% v 10%) in the past year. Sibling children are significantly more likely to be anaemic (42% v 32%) and to admit to depression (41% v 21%) or anxiety (45% v 37%). However, after adjusting for area, sex, and parental education levels only two differences remained: sibling children are more likely to be bullied (OR 1.5, 1.1-2.0; p = 0.006) and are less likely to confide in parents (OR 0.6, 0.3-0.8, p = 0.009). There were no significant differences in the key parameters between first and second born children. CONCLUSIONS: We found no detrimental effects of being an only child using the indicators measured. Being an only child may confer some benefits, particularly in terms of socialisation. PMID- 12765908 TI - Neurocysticercosis in West London. PMID- 12765909 TI - Apnoea and brain swelling in non-accidental head injury. AB - AIMS: (1) To identify whether infants and young children admitted to hospital with subdural haematomas (SDH) secondary to non-accidental head injury (NAHI), suffer from apnoea leading to radiological evidence of hypoxic ischaemic brain damage, and whether this is related to a poor prognosis; and (2) to determine what degree of trauma is associated with NAHI. METHODS: Retrospective case series (1992-98) with case control analysis of 65 children under 2 years old, with an SDH secondary to NAHI. Outcome measures were presenting symptoms, associated injuries and apnoea at presentation, brain swelling or hypoxic ischaemic changes on neuroimaging, and clinical outcome (KOSCHI). RESULTS: Twenty two children had a history of apnoea at presentation to hospital. Apnoea was significantly associated with hypoxic ischaemic brain damage. Severe symptoms at presentation, apnoea, and diffuse brain swelling/hypoxic ischaemic damage were significantly associated with a poor prognosis. Eighty five per cent of cases had associated injuries consistent with a diagnosis of non-accidental injury. CONCLUSIONS: Coma at presentation, apnoea, and diffuse brain swelling or hypoxic ischaemia all predict a poor outcome in an infant who has suffered from SDH after NAHI. There is evidence of associated violence in the majority of infants with NAHI. At this point in time we do not know the minimum forces necessary to cause NAHI. It is clear however that it is never acceptable to shake a baby. PMID- 12765910 TI - Height screening at school: ineffective without high standards and adequate resources. AB - BACKGROUND: The Coventry Consensus in 1998 recommended a single height measurement of all children at school entry or around the age of 5 years and prompt referral of children with height <0.4th centile for further assessment, in order to identify undetected and treatable asymptomatic growth disorders. AIM: To determine adherence and practicalities of following the Coventry Consensus recommendations in a community setting and the cost implications. METHODS: Anthropometric data of all children born between September 1992 and August 1993 in the Rhondda and Taff Ely area and measured in school year September 1998 to August 1999 were obtained from the National Child Health System (NCHS) and analysed in July 2000. RESULTS: Only 1592 (67.6%) of 2354 eligible children had their height measured. The NCHS could only flag up height data <2nd centile. Only five of the 15 children with height <0.4th centile were referred initially. Height measurements were not transcribed onto centiles in 75% of the case notes reviewed. When initially recalled, six of the 15 eligible children failed to attend the referral clinic. No new growth disorder was identified in any of these children. A conservative estimate of the cost to the health authority was pound 14 550 (US23 300 dollars; 20 500 euro) per annum. CONCLUSION: The study shows poor coverage and compliance together with a lack of parental awareness that short stature could be a potential health problem even in asymptomatic children. For a low yield programme to be successful and cost effective at the national level, a near 100% coverage is required. Further training of professionals in growth measurement and interpretation along with a campaign to raise both public and professional awareness is needed. PMID- 12765911 TI - Growth impairment in the very preterm and cognitive and motor performance at 7 years. AB - BACKGROUND: Infants born of low birth weight often have poor subsequent growth (especially if they were born very preterm), which has been shown to relate to later motor and cognitive development. AIMS: To assess a cohort of preterm infants at the age of 7 years for growth, motor, and cognitive measures, and investigate the effects of growth impairment on school performance. METHODS: A cohort of 280 children born before 32 completed weeks of gestation were tested, together with 210 term controls. RESULTS: Preterm children were significantly lighter and shorter than term controls and had smaller heads and lower body mass index (BMI). Median centiles for weight, height, head circumference, and BMI were 25, 25, 9, and 50 for boys and 50, 25, 9, and 50 for girls compared with 50, 50, 50, and 75 for controls. They performed significantly less well on all tests with a mean score of 91 (9.2) on the Developmental Test of Visual-Motor Integration, 89 (14.5) on the Wechsler-III IQ test, and 30.7% scoring at or below the 5th centile on the Movement Assessment Battery for Children. In boys, short stature and small heads were the best predictors of poor performance; in girls, a small head alone was a predictor for poor motor and cognitive performance. CONCLUSION: Poor postnatal growth in preterm infants, especially of the head, is associated with increased levels of motor and cognitive impairment at 7 years of age. This growth restriction appears to occur largely in the postnatal rather than antenatal period and may be amenable to intervention and subsequent improvement in outcome. PMID- 12765913 TI - Weaning of infants. AB - The WHO 2001 global recommendation is a one size fits all approach to weaning, an approach which may not take sufficient account of the special needs of some infants and fails to allow for the different problems encountered in the industrialised nations compared with economically developing countries. For the healthy normal birth weight full term infant born in an industrialised country, current research supports the benefit of exclusive breast milk feeding until 4-6 months. Evidence of harm through introducing solid food to these infants earlier than this is weak. Infants should be managed individually according to their needs. PMID- 12765915 TI - Same patient, different advice: a study into why doctors vary. AB - AIM: To understand why doctors differ in their recommendations in situations where there is little certainty about the long term outcomes of the possible treatment options. METHODS: A correlational design was used to examine the relation between preference for different treatment options and beliefs about likely outcomes for these options. Eighty doctors, with a mean of nine years in paediatric cardiology/surgery, attending a conference on serious congenital heart disease were studied. Main outcome measures were: ratings of the extent to which each of four treatment options were favoured; and subjective probabilities for three outcomes-death, survival with "good heart function" (New York Heart Association functional class (NYHA) I or II), and survival with "poor heart function" (NYHA III or IV)-for different treatment options over a 20 year time frame. RESULTS: Preference for one treatment option over another was most closely associated with the subjective estimate of the additional years with "good heart function" that it offered 10-20 years after surgery (Pearson's r = 0.66, p < 0.001). In influencing a preference, the possibility of early death was subordinate to optimising the late outcome. CONCLUSIONS: Doctors' treatment preferences are consistent with selecting the option that maximises the chance of the best outcome (long term survival with good heart function). Doctors' recommendations imply that they place more value on years of life in the child's far future than on life-years in the immediate future. PMID- 12765914 TI - Preservation of fertility in children treated for cancer. AB - As treatment for childhood cancer has become increasingly successful, adverse effects on reproductive function are assuming greater importance. Preservation of fertility before treatment must be considered in all young patients at high risk of infertility, and provision of such services requires collaboration between oncology centres and assisted conception units. The UK Children's Cancer Study Group is planning to audit current management of preservation of reproductive function in young patients with cancer, and the British Fertility Society is preparing a voluntary code of best practice to guide and inform clinicians and scientists. Limitation of radiation exposure by shielding of the testes and ovaries should be practiced where possible and sperm banking should be offered to all sexually mature boys at risk of infertility. The rapidly advancing experimental techniques for harvesting of gonadal tissue must be considered and embarked on without unrealistic expectations, although future utilisation of the tissue is unlikely to be realised until the next decade. PMID- 12765916 TI - Fluticasone propionate in asthma: a long term dose comparison study. AB - BACKGROUND: Few dose ranging studies have investigated optimal dosing with inhaled corticosteroids in children with asthma. AIMS: To compare the efficacy and tolerability of fluticasone propionate 100 or 200 microg twice daily in children with moderate to severe asthma for one year. METHODS: One year, randomised, double blind, parallel group, multicentre study. Children aged 4-11 years (n = 528) with moderate to severe asthma who had previously received high dose inhaled corticosteroids were given fluticasone propionate 100 or 200 microg twice daily for the 52 week treatment period. Efficacy (exacerbations, lung function, and symptoms) and tolerability (adverse events and cortisol levels) were measured. RESULTS: There was a non-significant decreased risk of experiencing an exacerbation at any time with fluticasone propionate 200 microg twice daily compared with fluticasone propionate 100 microg twice daily. This difference reached significance among patients with more severe asthma (defined by previous inhaled corticosteroid dose >800 microg/day). Daily record card morning peak expiratory flow (PEF) in the total population improved significantly more with the higher dose of fluticasone propionate (between group difference, weeks 1-52: 11.4 l/min). Clinic visit mean PEF improved from baseline with both doses, but the response was significantly greater with the higher dose (between group difference, week 52: 17.8 l/min). Both doses were equally well tolerated and overnight urinary cortisol concentrations were unchanged or slightly increased during treatment with either dose. CONCLUSION: This long term dose comparison study shows that treatment with fluticasone propionate 200 micro g twice daily may offer benefits over a lower dose, particularly in children with more severe asthma. PMID- 12765917 TI - Evaluation of vestibular functions in children with vertigo attacks. AB - AIM: To examine vestibular system functions in children with episodic vertigo attacks. METHODS: Thirty four children (20 males) aged 4-18 years with paroxysmal dizziness and/or vertigo attacks were evaluated. A medical history for vestibular symptoms and migraine was taken. Vestibular and auditory functions were assessed. RESULTS: Chronic headache attacks consistent with migraine were reported in 12 children and motion sickness was reported in 30. Family history in first degree relatives was positive for migraine in 29 children and for episodic vertigo in 22. Electronystagmography and videonystagmography showed two types of nystagmus: spontaneous vestibular nystagmus (41%) and benign paroxysmal positional nystagmus (BPPN) (59%). The first type of nystagmus was assessed as a sign of vestibulopathy and the patients with BPPN were diagnosed as having benign paroxysmal positional vertigo (BPPV). Audiometric examination in four cases revealed bilateral sensory neural hearing loss in low frequencies. Pure tone averages in 30 cases were within normal ranges; however low frequencies in 28 of them were approximately 10 dB lower than high frequencies. Unilateral caloric responses diminished in eight children. CONCLUSIONS: Peripheral vestibular problems in childhood present in a wide spectrum, which varies from a short episode of dizziness to a typical vestibular attack with fluctuating sensory neural hearing loss or episodes of BPPV. A considerable number of these vestibular problems might be related to the migraine syndrome. PMID- 12765918 TI - Fosfomycin for the initial treatment of acute haematogenous osteomyelitis. AB - BACKGROUND AND AIMS: At our institution there has been a dichotomous antimicrobial treatment behaviour for acute haematogenous osteomyelitis (AHOM) since 1984. The surgical department favoured fosfomycin as initial choice and the medical department beta lactams. We aimed to compare the performance of both strategies. METHODS: Data from patients discharged with the diagnosis of AHOM between January 1984 and January 1998 were gathered from the charts by means of a questionnaire. Patients receiving fosfomycin treatment (FT) were compared with those receiving fosfomycin plus other antimicrobials (FT+) and those receiving no fosfomycin treatment (NFT). RESULTS: A total of 103 patients aged 0.1-15.5 years (mean 6.5, median 6.9) with AHOM received no surgical treatment initially. In 23 (22.3%) FT was instilled initially, in 47 (45.6%) FT+, and in 33 (32.0%) NFT. The pathogen was established in 30%, 36%, and 42% of FT, FT+, and NFT patients, respectively, Staphylococcus aureus being the predominant isolate. Mean C reactive protein levels and erythrocyte sedimentation rates normalised in all treatment groups after two and four weeks, respectively. The mean duration of intravenous antimicrobial treatment in FT patients was 2.5 weeks, in FT+ patients 3.1 weeks, and in NFT patients 3.8 weeks (p < 0.05), whereas the mean duration of intravenous plus oral treatment was comparable (7.1 v 6.8 v 6.5 weeks). CONCLUSIONS: The leucocyte penetrating fosfomycin performed similarly to extracellular beta lactams in the treatment of AHOM. Intravenous treatment for longer than 2.5 weeks offered no advantage. PMID- 12765920 TI - Hypoparathyroidism and 22q11 deletion syndrome. AB - AIMS: To investigate a population of individuals with 22q11 deletion syndrome for hypocalcaemia. METHODS: A detailed clinical history enquiring into symptoms of hypocalcaemia and blood sampling to assess for hypocalcaemia and hypoparathyroidism, of patients outside the neonatal period known to have the 22q11 microdeletion from fluorescent in situ hybridisation studies was taken. RESULTS: Sixty one individuals were identified, of whom 23 were untraceable and one was unable to give informed consent. Biochemical investigations were performed on 27 subjects. Ten subjects had review of notes only. Four subjects had previously identified hypoparathyroidism. A new case of hypoparathyroidism was identified. Three subjects had borderline hypocalcaemia. DISCUSSION: In this population of patients with 22q11 deletion syndrome, 13% of the total or 30% of those biochemically assessed had evidence of reduced serum calcium concentrations. It is likely that 13-30% of patients with 22q11 deletion syndrome have possible hypoparathyroidism outside the neonatal period. Reported symptoms of hypocalcaemia did not correlate with biochemical evidence of persisting hypocalcaemia. We have shown that previously undiagnosed asymptomatic hypoparathyroidism occurs in patients with 22q11 deletion syndrome and conclude that screening of this population should be considered. PMID- 12765919 TI - Chronic urticaria: association with thyroid autoimmunity. AB - BACKGROUND: Though autoimmune phenomena have been regularly associated with chronic urticaria in adults, data in children are sparse. AIM: To describe our experience with children and adolescents with chronic urticaria and autoimmunity. METHODS AND RESULTS: Of 187 patients referred for evaluation of chronic urticaria during a 7.5 year period, eight (4.3%), all females aged 7-17 years, had increased levels of antithyroid antibody, either antithyroid peroxidase antibody (n = 4, >75 IU/ml), antithyroglobulin antibody (n = 2, >150 IU/ml), or both (n = 2). The duration of urticaria was four months to seven years. Five patients were euthyroid, one of whom was found to have increased antithyroid antibody levels five years after onset of the urticaria. One patient was diagnosed with Hashimoto thyroiditis three years before the urticaria, and was receiving treatment with thyroxine. Two other hypothyroid patients were diagnosed during the initial work up for urticaria (thyroxine 9.2 pmol/l, thyroid stimulating hormone (TSH) 40.2 mIU/l) and five years after onset of the urticaria (thyroxine 14 pmol/l, TSH 10.3 mIU/l). Both were treated with thyroxine but neither had remission of the urticaria. Five patients had a low positive titre of antinuclear antibodies. CONCLUSION: Children with chronic urticaria should be screened periodically for thyroxine, TSH, and antithyroid antibodies, as thyroid autoimmunity and hypothyroidism may appear several years after onset of the urticaria. PMID- 12765922 TI - 22q11 deletion: a multisystem disorder requiring multidisciplinary input. AB - AIM: To draw up recommendations for the investigation and management of children with a microdeletion of chromosome 22q11. METHODS: A retrospective review of case notes from patients with a chromosome 22q11 microdeletion identified by cytogenetics laboratories of the south and west of Britain over a four year period. RESULTS: A total of 210 cases were identified. Age at diagnosis was 0-1 years (34%), 1-4 (17%), 5-17 (35%), and 18 years or more (13%). School age children were less likely to be investigated than infants: echocardiography in school age 86% v in infancy 97%, serum calcium 66% v 89%, renal ultrasound scan 38% v 42%, lymphocyte count 26% v 68%, parental karyotype 78% v 88%. The yield of investigations remained high throughout all age groups with 42% of school age children shown to have hypocalcaemia and 25% abnormal findings on renal ultrasound. CONCLUSIONS: 22q11 microdeletion is a multisystem disorder requiring a set of core investigations at diagnosis. We recommend an echocardiogram, renal ultrasound scan, lymphocyte count and function, serum calcium, and parental karyotype as a minimum. Genetic counselling and community paediatric input is helpful for most families. PMID- 12765923 TI - Effect of a rapid influenza diagnosis. PMID- 12765924 TI - Safety of inhaled corticosteroids delivered by plastic and metal spacers. AB - BACKGROUND: Because of its non-electrostatic properties the metal Nebuchamber (NC) valved holding chamber (VHC) delivers a greater mass of aerosol to the mouth than the polypropylene Aerochamber (AC) VHC. Delivery of more aerosol to the lungs may also increase systemic absorption of inhaled corticosteroids (ICS) and hypothalamo-pituitary-adrenal (HPA) suppression. METHODS: Thirty children (mean 4.3 (SD 0.3) years) received 200 micro g budesonide twice daily by NC or AC, both with the mask provided, in a randomised, two month crossover trial. Twenty four hour urinary free cortisol (UFC) was determined as a measure of HPA suppression. RESULTS: UFC decreased from 42.3 (7.8) nmol UFC/nmol creatinine control to 26.2 (2.4) (p = 0.06 v control) after AC, and to 24.5 (2.5) (p = 0.04 v control) after NC (p = 0.4 AC v NC). CONCLUSIONS: Despite a greater total dose delivered to the mouth, NC is not associated with greater HPA suppression when using 400 micro g/day budesonide under real life conditions in young children. PMID- 12765925 TI - Systemic neutrophil activation in acute preschool viral wheeze. AB - BACKGROUND: In preschool children, attacks of wheeze are usually triggered by viral colds. The inflammatory substrate in preschool viral wheeze (PVW) is unclear, but epidemiological data suggest that most PVW is not caused by allergic inflammation. We therefore speculated that the neutrophils are an important effector cell. Systemic neutrophil activation is the first stage for the development of pulmonary neutrophilia. Markers of neutrophil activation are shedding of the adhesion molecule L-selectin from the cell surface, upregulation of Mac-1 expression, and an increase in serum soluble L-selectin. AIMS: To obtain evidence for systemic neutrophil activation during PVW. METHODS: Preschool children (1-5 years) admitted to hospital with acute PVW (n = 20) and normal controls (n = 18) were studied. Adhesion molecule expression on CD16 positive neutrophils was determined in both groups and expressed as molecules of equivalent fluorochrome (MEF). Serum soluble L-selectin was analysed by ELISA. RESULTS: Compared with controls, children with PVW had reduced neutrophil L selectin expression (median MEF (IQR): 69 (11 to 96) units versus 136 (109 to 163) units, p < 0.001) and higher serum soluble L-selectin (2.8 (2.3 to 3.1) versus 2.4 (2.2 to 2.6) micro g/ml, p = 0.04). There was no significant difference in neutrophil Mac-1 expression. CONCLUSION: Systemic neutrophil activation is associated with acute PVW. PMID- 12765926 TI - Bochdalek diaphragmatic hernia: not only a neonatal disease. AB - AIMS: To characterise the clinical manifestations of late presenting Bochdalek diaphragmatic hernia (DH), the incidence of misdiagnosis, and prognosis; and to explore the sequence of events that leads to this clinical picture. METHODS: Retrospective chart review. All children with Bochdalek DH were identified. Children 1 month of age and older at the time of diagnosis were included. RESULTS: Twenty two children with Bochdalek DH met the inclusion criteria. Three clinical presentations could be defined. Fourteen children presented with acute onset of symptoms, predominantly vomiting and respiratory distress. Four had chronic non-specific gastrointestinal or respiratory symptoms, and in four the DH was found incidentally. Although five children were initially misdiagnosed, in 20 children (91%) the correct diagnosis was made on x ray examination. One child experienced a complicated course when the x ray picture was misinterpreted as pneumothorax. All children had favourable outcome. Two children had previously normal chest imaging, suggesting acquired herniation. A large pleural effusion without DH in a 9.5 year old girl with an abdominal infection prior to presenting with herniation suggests a pre-existing defect in the diaphragm. CONCLUSIONS: Late presenting Bochdalek DH can present with acute or chronic gastrointestinal, or less frequently, respiratory symptoms. It can also be found incidentally. Misdiagnosis can result in significant morbidity. Favourable outcome is expected when the correct diagnosis is made. The sequence of events is probably herniation of abdominal viscera through a pre-existing diaphragmatic defect. Although rare, DH should be considered in any child presenting with respiratory distress or with symptoms suggestive of gastrointestinal obstruction. PMID- 12765928 TI - Thrombocytopenia and Plasmodium falciparum malaria in children with different exposures. AB - We studied thrombocytopenia during acute Plasmodium falciparum malaria in 64 traveller children from Paris (France), 85 children from Dakar (Senegal) with an intermittent exposure (69 with severe attack or cerebral malaria), and 81 children from Libreville (Gabon) with a perennial exposure (43 with severe attack or cerebral malaria). Initial thrombocytopenia was present in 43-58% of children with P falciparum malaria but was not more frequent in severe outcome or cerebral malaria. Low parasitaemia may lead to the misdiagnosis of malaria and delayed treatment when there is associated thrombocytopenia PMID- 12765927 TI - Chloramphenicol or ceftriaxone, or both, as treatment for meningitis in developing countries? AB - AIMS: To determine in children with meningitis whether there is any difference in mortality and neurological sequelae using chloramphenicol as first line treatment, with a change to ceftriaxone if chloramphenicol resistance is shown in vitro, compared to using ceftriaxone as first line treatment, with a change to chloramphenicol if there is no evidence of in vitro resistance. METHODS: An observational study with a retrospective control group nested within a randomised trial of fluid management for bacterial meningitis where clinical care was standardised. Chloramphenicol is standard treatment for bacterial meningitis in Papua New Guinea. In the first 150 cases we used chloramphenicol and only changed treatment to ceftriaxone if chloramphenicol resistance for cerebrospinal fluid isolates was proved. After finding 20% of Haemophilus influenzae were resistant to chloramphenicol, and that most affected children had poor outcomes, we changed to an alternative strategy. In the next 196 cases first line treatment was ceftriaxone and treatment was changed to chloramphenicol if the isolated bacteria were found to be susceptible. RESULTS: When chloramphenicol was used as first line treatment for meningitis followed by ceftriaxone when in vitro resistance was shown, there was invariably a very poor outcome in chloramphenicol resistant disease (71% of children died or had severe neurological complications). Using ceftriaxone as first line treatment was effective in reducing mortality and neurological sequelae from chloramphenicol resistant Haemophilus influenzae type (71% v 9%, relative risk 0.13; 95% CI 0.02 to 0.87; p = 0.013). Changing to chloramphenicol if there was no evidence of in vitro resistance was less than half the cost of empirical use of ceftriaxone for a full course for all children with meningitis. CONCLUSIONS: Using a third generation cephalosporin as first line treatment is effective in dealing with the problem of poor outcomes from meningitis due to Haemophilus influenzae that is resistant to chloramphenicol, and a strategy of changing to chloramphenicol if in vitro susceptibility is shown will reduce the use of expensive third generation cephalosporins without comprising on clinical outcomes. This highlights the urgent need to reduce the costs of third generation cephalosporins, to improve bacteriological services in developing countries, and to introduce effective and affordable vaccines against H influenzae and Streptococcus pneumoniae. PMID- 12765930 TI - The black hole in my CV. PMID- 12765929 TI - Pantoea agglomerans as a cause of septic arthritis after palm tree thorn injury; case report and literature review. AB - We report the case of a 14 year old healthy boy, who was admitted six weeks after being injured by a palm tree thorn, with limping caused by pain and swelling in his right knee. An ultrasound examination revealed a foreign body in the posterior lateral aspect of the right knee. Pantoea agglomerans was identified in the synovial fluid. The patient underwent two arthrotomies and was treated with amoxicillin-clavulanate intravenously for three weeks. The postoperative course was uneventful, and joint function returned to normal. A review of the literature between 1953 and 2002 revealed that bacterial growth after plant thorn injuries is reported infrequently. Yet when reported, Pantoea agglomearns is the most common organism found. Therefore, it must be considered and suspected in "aseptic" cases of arthritis, when there is a history of a plant thorn injury. We also emphasise the efficacy of ultrasound examination in these cases to identify the presence and location of a plant thorn. PMID- 12765931 TI - Once upon a time... AB - In this, the first of two articles discussing literature for and about children, we will be considering how writing for the young has changed, reflecting different and evolving perspectives on childhood. In the second article we will be asking whether literature can be used creatively and usefully in the training of doctors. The suggestion for the topic arose from a session we organised for paediatricians in the Communication and Management module of the MSc in Child Health at Leeds University. PMID- 12765932 TI - Not just an appendix: Sir Frederick Treves. AB - The history of the anatomy and surgery of the appendix is a beautiful chapter in medical education, and we appreciate the role of Sir Frederick Treves in its development. PMID- 12765933 TI - Improving mental health through parenting programmes: are the results valid? PMID- 12765934 TI - Vitamin K deficient bleeding in cystic fibrosis. PMID- 12765935 TI - Adrenal crisis due to inhaled steroids is underestimated. PMID- 12765936 TI - Cultural representation of newborn feeding. PMID- 12765937 TI - Moderately high doses still need to be considered for very young children. PMID- 12765938 TI - Differential effects of rosiglitazone on skeletal muscle and liver insulin resistance in A-ZIP/F-1 fatless mice. AB - To determine the role of adipocytes and the tissue-specific nature in the insulin sensitizing action of rosiglitazone, we examined the effects of 3 weeks of rosiglitazone treatment on insulin signaling and action during hyperinsulinemic euglycemic clamps in awake A-ZIP/F-1 (fatless), fat-transplanted fatless, and wild-type littermate mice. We found that 53 and 66% decreases in insulin stimulated glucose uptake and insulin receptor substrate (IRS)-1-associated phosphatidylinositol (PI) 3-kinase activity in skeletal muscle of fatless mice were normalized after rosiglitazone treatment. These effects of rosiglitazone treatment were associated with 50% decreases in triglyceride and fatty acyl-CoA contents in the skeletal muscle of rosiglitazone-treated fatless mice. In contrast, rosiglitazone treatment exacerbated hepatic insulin resistance in the fatless mice and did not affect already reduced IRS-2-associated PI 3-kinase activity in liver. The worsening of insulin action in liver was associated with 30% increases in triglyceride and fatty acyl-CoA contents in the liver of rosiglitazone-treated fatless mice. In conclusion, these data support the hypothesis that rosiglitazone treatment enhanced insulin action in skeletal muscle mostly by its ability to repartition fat away from skeletal muscle. PMID- 12765939 TI - Reduced activation of phosphatidylinositol-3 kinase and increased serine 636 phosphorylation of insulin receptor substrate-1 in primary culture of skeletal muscle cells from patients with type 2 diabetes. AB - To understand better the defects in the proximal steps of insulin signaling during type 2 diabetes, we used differentiated human skeletal muscle cells in primary culture. When compared with cells from control subjects, myotubes established from patients with type 2 diabetes presented the same defects as those previously evidenced in vivo in muscle biopsies, including defective stimulation of phosphatidylinositol (PI) 3-kinase activity, decreased association of PI 3-kinase with insulin receptor substrate (IRS)-1 and reduced IRS-1 tyrosine phosphorylation during insulin stimulation. In contrast to IRS-1, the signaling through IRS-2 was not altered. Investigating the causes of the reduced tyrosine phosphorylation of IRS-1, we found a more than twofold increase in the basal phosphorylation of IRS-1 on serine 636 in myotubes from patients with diabetes. Concomitantly, there was a higher basal mitogen-activated protein kinase (MAPK) activity in these cells, and inhibition of the MAPKs with PD98059 strongly reduced the level of serine 636 phosphorylation. These results suggest that IRS-1 phosphorylation on serine 636 might be involved in the reduced phosphorylation of IRS-1 on tyrosine and in the subsequent alteration of insulin-induced PI 3-kinase activation. Moreover, increased MAPK activity seems to play a role in the phosphorylation of IRS-1 on serine residue in human muscle cells. PMID- 12765940 TI - Fetal programming of perivenous glucose uptake reveals a regulatory mechanism governing hepatic glucose output during refeeding. AB - Increased hepatic gluconeogenesis maintains glycemia during fasting and has been considered responsible for elevated hepatic glucose output in type 2 diabetes. Glucose derived periportally via gluconeogenesis is partially taken up perivenously in perfused liver but not in adult rats whose mothers were protein restricted during gestation (MLP rats)-an environmental model of fetal programming of adult glucose intolerance exhibiting diminished perivenous glucokinase (GK) activity. We now show that perivenous glucose uptake rises with increasing glucose concentration (0-8 mmol/l) in control but not MLP liver, indicating that GK is flux-generating. The data demonstrate that acute control of hepatic glucose output is principally achieved by increasing perivenous glucose uptake, with rising glucose concentration during refeeding, rather than by downregulation of gluconeogenesis, which occurs in different hepatocytes. Consistent with these observations, glycogen synthesis in vivo commenced in the perivenous cells during refeeding, MLP livers accumulating less glycogen than controls. GK gene transcription was unchanged in MLP liver, the data supporting a recently proposed posttranscriptional model of GK regulation involving nuclear cytoplasmic transport. The results are pertinent to impaired regulation of hepatic glucose output in type 2 diabetes, which could arise from diminished GK mediated glucose uptake rather than increased gluconeogenesis. PMID- 12765941 TI - Glucose 6-phosphate regulates hepatic glycogenolysis through inactivation of phosphorylase. AB - High glucose concentration suppresses hepatic glycogenolysis by allosteric inhibition and dephosphorylation (inactivation) of phosphorylase-a. The latter effect is attributed to a direct effect of glucose on the conformation of phosphorylase-a. Although glucose-6-phosphate (G6P), like glucose, stimulates dephosphorylation of phosphorylase-a by phosphorylase phosphatase, its physiological role in regulating glycogenolysis in intact hepatocytes has not been tested. We show in this study that metabolic conditions associated with an increase in G6P, including glucokinase overexpression and incubation with octanoate or dihydroxyacetone, cause inactivation of phosphorylase. The latter conditions also inhibit glycogenolysis. The activity of phosphorylase-a correlated inversely with the G6P concentration within the physiological range. The inhibition of glycogenolysis and inactivation of phosphorylase-a caused by 10 mmol/l glucose can be at least in part counteracted by inhibition of glucokinase with 5-thioglucose, which lowers G6P. In conclusion, metabolic conditions that alter the hepatic G6P content affect glycogen metabolism not only through regulation of glycogen synthase but also through regulation of the activation state of phosphorylase. Dysregulation of G6P in diabetes by changes in activity of glucokinase or glucose 6-phosphatase may be a contributing factor to impaired suppression of glycogenolysis by hyperglycemia. PMID- 12765942 TI - Insulin resistance and lipodystrophy in mice lacking ribosomal S6 kinase 2. AB - The p90 ribosomal S6 kinase 2 (RSK2) is a serine/threonine kinase with high expression levels in adipose tissue. Numerous in vitro studies show that RSK2 is activated by a broad number of cellular stimuli and suggest that RSK2 is involved in the regulation of a variety of cellular processes. However, the physiological role of RSK2 still remains elusive. We therefore generated rsk2 knockout (KO) mice to better understand the function of RSK2 in vivo. Birth weights of RSK2 KO mice are normal, but the body weight is reduced with age, as compared with wild type littermates. We found that the difference in body weight was largely caused by a specific loss of white adipose tissue that is accompanied by reduced serum levels of the adipocyte-derived peptide, leptin. KO mice also have impaired glucose tolerance and elevated fasting insulin and glucose levels that are restored following administration of low amounts of leptin, which do not affect food intake. We conclude that RSK2 plays a novel and an important role in regulation of adipose mass in mice and speculate that the reduction in fat tissue may negatively affect insulin sensitivity, as observed in human lipodystrophy, through reduced levels of adipocyte-derived factors, such as leptin. PMID- 12765943 TI - Combined infusion of epinephrine and norepinephrine during moderate exercise reproduces the glucoregulatory response of intense exercise. AB - Intense exercise (IE) (>80% O(2max)) causes a seven- to eightfold increase in glucose production (R(a)) and a fourfold increase in glucose uptake (R(d)), resulting in hyperglycemia, whereas moderate exercise (ME) causes both to double. If norepinephrine (NE) plus epinephrine (Epi) infusion during ME produces the plasma levels and R(a) of IE, this would prove them capable of mediating these responses. Male subjects underwent 40 min of 53% O(2max) exercise, eight each with saline (control [CON]), or with combined NE + Epi (combined catecholamine infusion [CCI]) infusion from min 26-40. In CON and CCI, NE levels reached 7.3 +/ 0.7 and 33.1 +/- 2.9 nmol/l, Epi 0.94 +/- 0.08 and 7.06 +/- 0.44 nmol/l, and R(a) 3.8 +/- 0.4 and 12.9 +/- 0.8 mg. kg(-1). min(-1) (P < 0.001), respectively, at 40 min. R(d) increased to 3.5 +/- 0.4 vs. 11.2 +/- 0.8 mg. kg(-1). min(-1) and glycemia 5.2 +/- 0.2 mmol/l in CON vs. 6.5 +/- 0.2 mmol/l in CCI (P < 0.001). The glucagon-to-insulin ratio did not differ. Comparing CCI data to those from 14-min IE (n = 16), peak NE (33.6 +/- 5.1 nmol/l), Epi (5.32 +/- 0.93 nmol/l), and R(a) (13.0 +/- 1.0 mg. kg(-1). min(-1)) were comparable. The induced increments in NE, Epi, and R(a), all of the same magnitude as in IE, strongly support that circulating catecholamines can be the prime regulators of R(a) in IE. PMID- 12765944 TI - Involvement of AMP-activated protein kinase in glucose uptake stimulated by the globular domain of adiponectin in primary rat adipocytes. AB - Adiponectin is an abundant adipocyte-derived plasma protein with anti atherosclerotic and insulin-sensitizing properties that suppresses hepatic glucose production and enhances glucose uptake into skeletal muscle. To characterize the potential effects of adiponectin on glucose uptake into adipose cells, we incubated isolated epididymal rat adipocytes with the globular domain of recombinant adiponectin purified from an E. coli expression system. Globular adiponectin increased glucose uptake in adipocytes without stimulating tyrosine phosphorylation of the insulin receptor or insulin receptor substrate-1, and without enhancing phosphorylation of Akt on Ser-473. Globular adiponectin further enhanced insulin-stimulated glucose uptake at submaximal insulin concentrations and reversed the inhibitory effect of tumor necrosis factor-alpha on insulin stimulated glucose uptake. Cellular treatment with globular adiponectin increased the Thr-172 phosphorylation and catalytic activity of AMP-activated protein kinase and enhanced the Ser-79 phosphorylation of acetyl CoA carboxylase, an enzyme downstream of AMP kinase in adipose cells. Inhibition of AMP kinase activation using two pharmacological inhibitors (adenine 9-beta-D arabinofuranoside and compound C) completely abrogated the increase in glucose uptake stimulated by globular adiponectin, indicating that AMP kinase is integrally involved in the adiponectin signal transduction pathway. Coupled with recent evidence that the effects of adiponectin are mediated via AMP kinase activation in liver and skeletal muscle, the findings reported here provide an important mechanistic link in the signaling effects of adiponectin in diverse metabolically responsive tissues. PMID- 12765945 TI - Pioglitazone reduces hepatic fat content and augments splanchnic glucose uptake in patients with type 2 diabetes. AB - The effect of pioglitazone on splanchnic glucose uptake (SGU), endogenous glucose production (EGP), and hepatic fat content was studied in 14 type 2 diabetic patients (age 50 +/- 2 years, BMI 29.4 +/- 1.1 kg/m(2), HbA(1c) 7.8 +/- 0.4%). Hepatic fat content (magnetic resonance spectroscopy) and SGU (oral glucose load- insulin clamp technique) were quantitated before and after pioglitazone (45 mg/day) therapy for 16 weeks. Subjects received a 7-h euglycemic insulin (100 mU. m(-2). min(-1)) clamp, and a 75-g oral glucose load was ingested 3 h after starting the insulin clamp. Following glucose ingestion, the steady-state glucose infusion rate during the insulin clamp was decreased appropriately to maintain euglycemia. SGU was calculated by subtracting the integrated decrease in glucose infusion rate during the 4 h after glucose ingestion from the ingested glucose load. 3-[(3)H]glucose was infused during the initial 3 h of the insulin clamp to determine rates of EGP and glucose disappearance (R(d)). Pioglitazone reduced fasting plasma glucose (10.0 +/- 0.7 to 7.5 +/- 0.6 mmol/l, P < 0.001) and HbA(1c) (7.8 +/- 0.4 to 6.7 +/- 0.3%, P < 0.001) despite increased body weight (83 +/- 3 to 86 +/- 3 kg, P < 0.001). During the 3-h insulin clamp period before glucose ingestion, pioglitazone improved R(d) (6.9 +/- 0.5 vs. 5.2 +/- 0.5 mg. kg(-1). min(- 1), P < 0.001) and insulin-mediated suppression of EGP (0.21 +/- 0.04 to 0.06 +/- 0.02 mg. kg(-1). min(-1), P < 0.01). Following pioglitazone treatment, hepatic fat content decreased from 19.6 +/- 3.6 to 10.4 +/- 2.1%, (P < 0.005), and SGU increased from 33.0 +/- 2.8 to 46.2 +/- 5.1% (P < 0.005). Pioglitazone treatment in type 2 diabetes 1) decreases hepatic fat content and improves insulin-mediated suppression of EGP and 2) augments splanchnic and peripheral tissue glucose uptake. Improved splanchnic/peripheral glucose uptake and enhanced suppression of EGP contribute to the improvement in glycemic control in patients with type 2 diabetes. PMID- 12765946 TI - Starvation and diabetes reduce the amount of pyruvate dehydrogenase phosphatase in rat heart and kidney. AB - The pyruvate dehydrogenase complex (PDC) is inactivated in many tissues during starvation and diabetes to conserve three-carbon compounds for gluconeogenesis. This is achieved by an increase in the extent of PDC phosphorylation caused in part by increased pyruvate dehydrogenase kinase (PDK) activity due to increased PDK expression. This study examined whether altered pyruvate dehydrogenase phosphatase (PDP) expression also contributes to changes in the phosphorylation state of PDC during starvation and diabetes. Of the two PDP isoforms expressed in mammalian tissues, the Ca(2+)-sensitive isoform (PDP1) is highly expressed in rat heart, brain, and testis and is detectable but less abundant in rat muscle, lung, kidney, liver, and spleen. The Ca(2+)-insensitive isoform (PDP2) is abundant in rat kidney, liver, heart, and brain and is detectable in spleen and lung. Starvation and streptozotocin-induced diabetes cause decreases in PDP2 mRNA abundance, PDP2 protein amount, and PDP activity in rat heart and kidney. Refeeding and insulin treatment effectively reversed these effects of starvation and diabetes, respectively. These findings indicate that opposite changes in expression of specific PDK and PDP isoenzymes contribute to hyperphosphorylation and therefore inactivation of the PDC in heart and kidney during starvation and diabetes. PMID- 12765947 TI - Differential regulation of protein dynamics in splanchnic and skeletal muscle beds by insulin and amino acids in healthy human subjects. AB - To determine the in vivo effect of amino acids (AAs) alone or in combination with insulin on splanchnic and muscle protein dynamics, we infused stable isotope tracers of AAs in 36 healthy subjects and sampled from femoral artery and vein and hepatic vein. The subjects were randomized into six groups and were studied at baseline and during infusions of saline (group 1), insulin (0.5 mU. kg(-1). min(-1)) (group 2), insulin plus replacement of AAs (group 3) insulin plus high dose AAs (group 4), or somatostatin and baseline replacement doses of insulin, glucagon and GH plus high dose of AAs (group 5) or saline (group 6). Insulin reduced muscle release of AAs mainly by inhibition of protein breakdown. Insulin also enhanced AA-induced muscle protein synthesis (PS) and reduced leucine transamination. The main effect of AAs on muscle was the enhancement of PS. Insulin had no effect on protein dynamics or leucine transamination in splanchnic bed. However, AAs reduced protein breakdown and increased synthesis in splanchnic bed in a dose-dependent manner. AAs also enhanced leucine transamination in both splanchnic and muscle beds. Thus insulin's anabolic effect was mostly on muscle, whereas AAs acted on muscle as well as on splanchnic bed. Insulin achieved anabolic effect in muscle by inhibition of protein breakdown, enhancing AA induced PS, and reducing leucine transamination. AAs largely determined protein anabolism in splanchnic bed by stimulating PS and decreasing protein breakdown. PMID- 12765948 TI - Renal compensation for impaired hepatic glucose release during hypoglycemia in type 2 diabetes: further evidence for hepatorenal reciprocity. AB - During liver transplantation and after both meal ingestion and prolonged fasting, renal glucose release (RGR) increases while hepatic glucose release (HGR) decreases. These and other observations have led to the concept of hepatorenal reciprocity. According to this concept, reciprocal changes in hepatic and renal glucose release may occur to minimize deviations from normal glucose homeostasis. We further assessed this concept by testing the hypothesis that during counterregulation of hypoglycemia in patients with type 2 diabetes, who would be expected to have reduced HGR, RGR would be increased. Accordingly, we performed hypoglycemic hyperinsulinemic clamp experiments (approximately 3.1 mmol/l) in 12 type 2 diabetic and in 10 age-weight-matched nondiabetic volunteers and measured total endogenous glucose release (TEGR) and RGR using a combined isotopic net balance approach. HGR was calculated as the difference between TEGR and RGR since only these organs are capable of releasing glucose. We found that during comparable hypoglycemia and hyperinsulinemia, TEGR was reduced in type 2 diabetes (6.6 +/- 0.6 vs. 10.2 +/- 1.1 micromol. kg(-1). min(-1) in nondiabetic volunteers, P = 0.01) due to reduced HGR (3.9 +/- 0.5 vs. 8.6 +/- 1.0 micromol. kg(-1). min(-1) in nondiabetic volunteers, P = 0.0015). In contrast, RGR was increased approximately twofold in type 2 diabetes (3.3 +/- 0.5 vs. 1.6 +/- 0.3 micromol. kg(-1). min(-1) in nondiabetic volunteers, P = 0.015). Plasma epinephrine, lactate, and free fatty acid concentrations, which would promote RGR, were also greater in type 2 diabetes (all P < 0.01). Our results provide further support for hepatorenal reciprocity and may explain at least in part the relatively low occurrence of severe hypoglycemia in type 2 diabetes compared with type 1 diabetes where both HGR and RGR counterregulatory responses are reduced. PMID- 12765950 TI - Low circulating IGF-II concentrations predict weight gain and obesity in humans. AB - Results from experimental and gene-association studies suggest that IGF-II may influence body weight regulation and that individuals with low IGF-II levels may be more susceptible to weight gain and obesity. We therefore assessed the association between circulating concentrations of IGF-II and subsequent weight gain and progression to obesity. Participants in this study were 463 nonobese men and women aged between 45 and 60 years with normal glucose tolerance and with metabolic and anthropometric assessments at baseline and follow-up clinic visits. We examined the association between baseline concentrations of fasting serum IGF II and risk of gaining > or =2.5 kg body wt or developing obesity using unconditional logistic regression. A total of 217 participants gained > or =2.5 kg body wt, and 29 developed obesity after >4 years of follow-up. In multivariate analysis, baseline IGF-II levels were significantly lower in participants who subsequently gained weight compared with individuals who remained stable or lost weight (P = 0.010). Similarly, individuals who developed obesity had lower baseline IGF-II levels (P = 0.006). Relatively higher IGF-II levels were also associated with a reduced risk of gaining weight (P for trend across quintiles of IGF-II = 0.006). Our data suggest that circulating IGF-II levels may play a role in body weight regulation and development of obesity in men and women with normal glucose tolerance. PMID- 12765949 TI - Increased phosphorylation of skeletal muscle glycogen synthase at NH2-terminal sites during physiological hyperinsulinemia in type 2 diabetes. AB - In type 2 diabetes, insulin activation of muscle glycogen synthase (GS) is impaired. This defect plays a major role for the development of insulin resistance and hyperglycemia. In animal muscle, insulin activates GS by reducing phosphorylation at both NH(2)- and COOH-terminal sites, but the mechanism involved in human muscle and the defect in type 2 diabetes remain unclear. We studied the effect of insulin at physiological concentrations on glucose metabolism, insulin signaling and phosphorylation of GS in skeletal muscle from type 2 diabetic and well-matched control subjects during euglycemic hyperinsulinemic clamps. Analysis using phospho-specific antibodies revealed that insulin decreases phosphorylation of sites 3a + 3b in human muscle, and this was accompanied by activation of Akt and inhibition of glycogen synthase kinase 3alpha. In type 2 diabetic subjects these effects of insulin were fully intact. Despite that, insulin-mediated glucose disposal and storage were reduced and activation of GS was virtually absent in type 2 diabetic subjects. Insulin did not decrease phosphorylation of sites 2 + 2a in healthy human muscle, whereas in diabetic muscle insulin infusion in fact caused a marked increase in the phosphorylation of sites 2 + 2a. This phosphorylation abnormality likely caused the impaired GS activation and glucose storage, thereby contributing to skeletal muscle insulin resistance, and may therefore play a pathophysiological role in type 2 diabetes. PMID- 12765951 TI - Leptin activation of corticosterone production in hepatocytes may contribute to the reversal of obesity and hyperglycemia in leptin-deficient ob/ob mice. AB - Glucocorticoids have been implicated as pathophysiological mediators of obesity and insulin resistance and are regulated by 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1). This enzyme regenerates active corticosterone from inactive 11-keto forms. To assess the role of 11beta-HSD1-mediated synthesis of active corticosterone in leptin-related obesity and diabetes, we examined the peripheral effect of leptin on 11beta-HSD1 activity and gene expression in vivo and in vitro in hepatocytes from ob/ob mice and in liver of streptozotocin (STZ)-treated ob/ob mice. We observed an inverse relationship between hepatic 11beta-HSD1 expression and body weight in ob/ob mice and lean littermates. Leptin treatment of ob/ob mice markedly increased hepatic 11beta-HSD1 activity and mRNA expression. This induction of 11beta-HSD1 expression corresponded to reduced levels of circulating corticosterone and weight loss in ob/ob mice treated with leptin, indicating that impaired hepatic 11beta-HSD1 expression may contribute to the pathogenesis of obesity in ob/ob mice. In addition, leptin treatment of STZ-treated ob/ob mice caused marked increases in hepatic 11beta-HSD1 levels associated with decreased body weight and a significant reduction in hyperglycemia due to pancreatic beta cell damage. Addition of leptin to ob/ob mouse primary hepatocytes led to a dose dependent increase in 11beta-HSD1 mRNA expression. In contrast, leptin did not influence 11beta-HSD1 expression in primary hepatocytes from db/db mice, indicating that leptin regulation of 11beta-HSD1 expression is probably mediated by the functional leptin receptor. Thus, leptin appears to be an important metabolic signal that directly activates intrahepatic corticosterone production. These findings suggest that the liver-specific interaction of leptin with 11beta HSD1 is involved in the development of obesity and insulin resistance in ob/ob mice. PMID- 12765952 TI - The presence of a catalytically inactive form of hormone-sensitive lipase is associated with decreased lipolysis in abdominal subcutaneous adipose tissue of obese subjects. AB - Hormone-sensitive lipase (HSL)-L is a key enzyme in the mobilization of fatty acids from triglyceride stores in adipocytes. A shorter variant of HSL (HSL-S) was detected in humans. This one is generated through in-frame skipping of exon 6 during the processing of HSL mRNA and results in a protein devoid of lipase activity. The role of HSL-S is unknown. The aims of this study were to identify both HSL variants in adipose tissue biopsies and to determine if the presence of HSL-S is correlated to the lipolytic capacity of adipocytes. The study was performed in human abdominal subcutaneous adipocytes from two groups of seven obese subjects. In the group of subjects with both HSL proteins (L+S) group, two immunoreactive bands (80 and 88 kDa) were detected, whereas only the 88-kDa protein was detected in the group with only the wild-type HSL-protein (L group). In the L+S group, the HSL activity was 20% lower (P < 0.05) and the (S/S(+)) HSL mRNA ratio was twofold higher than in the L group (P < 0.05). The maximally lipolytic capacities measured from isolated adipocytes incubated with norepinephrine or other lipolytic agents were 40% lower in the L+S group (P < 0.05). These results suggest that the presence of the truncated HSL protein is associated with an impaired adipocyte lipolysis. PMID- 12765953 TI - Discrete and complementary mechanisms of protection of beta-cells against cytokine-induced and oxidative damage achieved by bcl-2 overexpression and a cytokine selection strategy. AB - We have been investigating the potential utility of engineered cell lines as surrogates for primary islet cells in treatment of type 1 diabetes. To this end, two strategies that have emerged for procuring cell lines with resistance to immune-mediated damage are 1) selection of cytokine-resistant cell lines by growth of INS-1 insulinoma cells in iteratively increasing concentrations of interleukin (IL)-1beta + gamma-interferon (IFN-gamma), and 2) stable overexpression of the anti-apoptotic gene bcl-2 in INS-1 cells. Herein, we show that bcl-2-overexpressing cells are resistant to the cytotoxic effects of reactive oxygen and nitrogen species (ROS/RNS), but are only modestly protected against high concentrations of IL-1beta + INF-gamma, whereas the converse is true in cytokine selected cells. We also found that the combination of bcl-2 expression and cytokine selection confers a broader spectrum of resistance than either procedure alone, such that the resultant cells are highly resistant to cytokines and ROS/RNS, with no impairment in glucose-stimulated insulin secretion. INS-1-derived cells with combined bcl-2 expression and cytokine selection are also more resistant to damage induced by coculture with mitogen activated peripheral blood mononuclear cells. Surprisingly, application of the cytokine selection procedure to bcl-2-overexpressing cells does not result in impairment of nuclear factor-kappaB translocation, iNOS expression, and NO production, as clearly occurs upon application of the selection procedure to cells without bcl-2 overexpression. Further investigation of the diverse pathways involved in the development of cytokine and ROS/RNS resistance may define simplified and specific strategies for preservation of beta-cell mass. PMID- 12765954 TI - The degree of phylogenetic disparity of islet grafts dictates the reliance on indirect CD4 T-cell antigen recognition for rejection. AB - Cellular xenograft rejection involves a pronounced contribution of CD4 T-cells recognizing antigens in association with recipient MHC class II molecules. However, the requirement for such "indirect" antigen recognition for acute islet xenograft is not clear, especially as a function of the phylogenetic disparity between the donor and recipient species. In vitro studies show that C57BL/6 (B6) mouse T-cells respond directly to either allogeneic BALB/c or phylogenetically related xenogeneic WF rat stimulator cells while having undetectable responses to phylogenetically disparate porcine stimulator cells. Although all types of grafts rejected acutely in wild-type mice, this response demonstrated markedly differing dependence on host MHC class II antigen presentation, depending on the donor species. While BALB/c islet allografts were acutely rejected in B6 MHC class II deficient (C2D) recipients, WF rat xenografts demonstrated marked prolongation in C2D hosts relative to wild-type recipients. Interestingly, neonatal porcine islet (NPI) xenografts uniformly survived long term (>100 days) in untreated C2D hosts despite transfer of wild-type CD4 T-cells, demonstrating that survival in C2D recipients was not secondary to a lack of CD4 T-cells seen in such mice. Taken together, these results show a marked hierarchy in the requirement for host MHC class II-restricted indirect pathway in the rejection of pancreatic islet grafts. Thus, while cellular rejection of porcine xenografts is generally quite vigorous, this pathway is relatively finite, displaying a major reliance on host MHC class II-dependent antigen presentation for acute rejection. PMID- 12765955 TI - Fetal or neonatal low-glycotoxin environment prevents autoimmune diabetes in NOD mice. AB - Advanced glycation end products (AGEs) are implicated in beta-cell oxidant stress. Diet-derived AGE (dAGE) are shown to contribute to end-organ toxicity attributed to diabetes. To assess the role of dAGE on type 1 diabetes, NOD mice were exposed to a high-AGE diet (H-AGE) and to a nutritionally similar diet with approximate fivefold-lower levels of N(epsilon)-carboxymethyllysine (CML) and methylglyoxal-derivatives (MG) (L-AGE). Suppression of serum CML and MG in L-AGE fed mice was marked by suppression of diabetes (H-AGE mice >94% vs. L-AGE mice 33% in founder [F](0), 14% in F(1), and 13% in F(2) offspring, P < 0.006) and by a delay in disease onset (4-month lag). Survival for L-AGE mice was 76 vs. 0% after 44 weeks of H-AGE mice. Reduced insulitis in L-AGE versus H-AGE mice (P < 0.01) was marked by GAD- and insulin-unresponsive pancreatic interleukin (IL)-4 positive CD4+ cells compared with the GAD- and insulin-responsive interferon (IFN)-gamma-positive T-cells from H-AGE mice (P < 0.005). Splenocytes from L-AGE mice consisted of GAD- and insulin-responsive IL-10-positive CD4+ cells compared with the IFN-gamma-positive T-cells from H-AGE mice (P < 0.005). Therefore, high AGE intake may provide excess antigenic stimulus for T-cell-mediated diabetes or direct beta-cell injury in NOD mice; both processes are ameliorated by maternal or neonatal exposure to L-AGE nutrition. PMID- 12765956 TI - Increased vulnerability of brain mitochondria in diabetic (Goto-Kakizaki) rats with aging and amyloid-beta exposure. AB - This study evaluated the respiratory indexes (respiratory control ratio [RCR] and ADP/O ratio), mitochondrial transmembrane potential (DeltaPsim), repolarization lag phase, repolarization level, ATP/ADP ratio, and induction of the permeability transition pore of brain mitochondria isolated from normal Wistar and GK diabetic rats of different ages (1.5, 12, and 24 months of age). The effect of amyloid beta-peptides, 50 micromol/l Abeta(25-35) or 2 micromol/l Abeta(1-40), on mitochondrial function was also analyzed. Aging of diabetic mice induced a decrease in brain mitochondrial RCR, ADP/O, and ATP/ADP ratios but induced an increase in the repolarization lag phase. Brain mitochondria from older diabetic rats were more prone to the induction of the permeability transition pore, i.e., mitochondria from 24-month-old diabetic rats accumulated much less Ca(2+) (20 micromol/l) than those isolated from 12-month-old rats (50 micromol/l) or 1.5 month-old rats (100 micromol/l). In the presence of 50 micromol/l Abeta(25-35) or 2 micromol/l Abeta(1-40), age-related mitochondrial effects were potentiated. These results indicate that diabetes-related mitochondrial dysfunction is exacerbated by aging and/or by the presence of neurotoxic agents such as amyloid beta-peptides, supporting the idea that diabetes and aging are risk factors for the neurodegeneration induced by these peptides. PMID- 12765957 TI - Pancreatic beta-cells express phagocyte-like NAD(P)H oxidase. AB - The presence of a phagocyte-like NAD(P)H oxidase in pancreatic beta-cells was investigated. Three NAD(P)H oxidase components were found in pancreatic islets by RT-PCR: gp91(PHOX), p22(PHOX), and p47(PHOX). The components p67(PHOX) and p47(PHOX) were also demonstrated by Western blotting. Through immunohistochemistry, p47(PHOX) was mainly found in the central area of the islet, confirming the expression of this component by insulin-producing cells. Activation of NAD(P)H oxidase complex in the beta-cells was also examined by immunohistochemistry. The pancreatic islets presented slower kinetics of superoxide production than HIT-T15 cells, neutrophils, and macrophages, but they reached 66% that of the neutrophil nitroblue tetrazolium (NBT) reduction after 2 h of incubation. Glucose (5.6 mmol/l) increased NBT reduction by 75% when compared with control. The involvement of protein kinase C (PKC) in the stimulatory effect of glucose was confirmed by incubation of islets with phorbol myristate acetate (a PKC activator) and bysindoylmaleimide (GF109203X) (a PKC specific inhibitor). Diphenylene iodonium [an NAD(P)H oxidase inhibitor] abolished the increase of NBT reduction induced by glucose, confirming the NAD(P)H oxidase activity in pancreatic islets. Because reactive oxygen species are involved in intracellular signaling, the phagocyte-like NAD(P)H oxidase activation by glucose may play an important role for beta-cell functioning. PMID- 12765958 TI - Circadian variation in the onset of myocardial infarction: effect of duration of diabetes. AB - There are conflicting reports regarding circadian variation in the onset of acute myocardial infarction (MI) among patients with diabetes. We therefore, studied the circadian pattern of the incidence of acute MI in patients (n = 3,882) who were enrolled in the Onset Study stratified by the presence, type, and duration of diabetes. The Onset Study was conducted at 64 U.S. medical centers between August 1989 and September 1996. We used harmonic regression model to evaluate the circadian variation of MI symptom onset in patients with and without diabetes. Subgroup analysis was performed according to the presence, type, and duration of diabetes by the chi(2) test (dividing the day into four 6-h intervals). Patients without diabetes exhibited a prominent morning peak in the incidence of acute MI symptom onset (P < 0.001). In contrast, patients with type 1 diabetes and type 2 diabetes > or =5 years had a marked attenuation of the morning peak. Patients who had type 2 diabetes diagnosed within the previous 5 years had a pattern of onset of acute MI similar to patients without diabetes. Patients with type 1 diabetes and those with type 2 diabetes > or =5 years have an attenuation of the morning peak in acute MI. Inconsistency in observation of such an effect in patients with diabetes in the past may well have been due to difference in the duration of diabetes and thus the variable extent of underlying autonomic dysfunction. PMID- 12765959 TI - Mechanisms of abnormal cardiac repolarization during insulin-induced hypoglycemia. AB - Prolonged cardiac repolarization causes fatal cardiac arrhythmias. There is evidence that these contribute to sudden death associated with nocturnal hypoglycemia in young people with diabetes. We measured cardiac repolarization (QT interval [QTc] and QT dispersion [QTd]) during experimental hypoglycemia with and without beta-blockade and potassium infusion to establish possible mechanisms. Two groups of 10 nondiabetic men (study 1 and study 2) each underwent four hyperinsulinemic clamps: two euglycemic (5 mmol/l) and two hypoglycemic (5 mmol/l and 2.5 mmol/l for 60 min each). Study 1 was performed with and without potassium infusion to maintain normal concentrations and study 2 with and without beta-blockade (atenolol, 100 mg/day for 7 days). QTd was unchanged during euglycemia but increased during hypoglycemia (55 ms, P < 0.0001 vs. baseline), which was prevented by potassium (6 ms, P = 0.78). QTc increased significantly during hypoglycemia alone (67 ms, P < 0.0001) and during potassium replacement (46 ms, P = 0.02). In study 2, the increase in QTd during hypoglycemia (68 ms, P < 0.0001) was prevented by beta-blockade (3 ms, P = 0.88). The increase in QTc during hypoglycemia (55 ms, P < 0.0001) was prevented by beta-blockade (1 ms, P = 0.98). Our data indicate that hypoglycemia causes an acquired long QT syndrome. Sympathoadrenal stimulation is the main cause, through mechanisms that involve but are not limited to catecholamine-mediated hypokalemia. These abnormalities are prevented by selective beta-blockade. PMID- 12765960 TI - The natural history of progression from normal glucose tolerance to type 2 diabetes in the Baltimore Longitudinal Study of Aging. AB - The natural history of progression from normal glucose tolerance (NGT) to impaired fasting glucose (IFG), impaired glucose tolerance (IGT), and type 2 diabetes is not well defined. We studied this progression using biennial oral glucose tolerance tests performed in the Baltimore Longitudinal Study of Aging and survival analysis to assess progression from NGT to abnormal fasting plasma glucose (FPG; > or =6.1 mmol/l), abnormal 2-h plasma glucose (2hPG; > or =7.8 mmol/l), IFG (FPG 6.1-6.9 mmol/l, 2hPG < or =7.8 mmol/l), and IGT (FPG <6.1 mmol/l, 2hPG 7.8-11.0 mmol/l), and from IFG-IGT to diabetes (FPG > or =7.0 mmol/l or 2hPG > or =11.1 mmol/l). At baseline, the 815 subjects had a mean age of 57 years, 35% were women, and 60% had NGT. Of the 488 subjects with NGT, over half were followed for at least 10 years. By 10 years, 14% had progressed to abnormal FPG and 48% to abnormal 2hPG. Of the 267 subjects who progressed to IFG-IGT, 216 had additional follow-up. By 10 years, 8% of these progressed to diabetes by FPG whereas 27% progressed by 2hPG. In subsidiary analyses, we defined "abnormal" FPG as > or =5.55 mmol/l and "diabetic" FPG as > or =6.1 mmol/l, making the baseline prevalence of IFG similar to that of IGT. By these criteria, 43% progressed to abnormal FPG and 43% to abnormal 2hPG by 10 years of follow-up; among subjects developing impaired FPG or 2hPG, 22% progressed to diabetes by FPG whereas 17% progressed by 2hPG at 10 years. Nonetheless, 42% of subjects developing abnormal FPG did not develop abnormal 2hPG, and vice versa. We conclude that, although phenotypic differences in rates of progression are partly a function of diagnostic thresholds, fasting and postchallenge hyperglycemia may represent phenotypes with distinct natural histories in the evolution of type 2 diabetes. PMID- 12765961 TI - Free fatty acid-mediated impairment of glucose-stimulated insulin secretion in nondiabetic Oji-Cree individuals from the Sandy Lake community of Ontario, Canada: a population at very high risk for developing type 2 diabetes. AB - The Oji-Cree population of the Sandy Lake region of Ontario, Canada, has the third highest prevalence of type 2 diabetes in the world. Changes in their diet and physical activity over the past half-century, particularly the marked increase in consumption of dietary fats, are felt to be important factors accounting for this epidemic. The aim of the present study was to examine the beta-cell response to a 48-h approximately twofold elevation of plasma free fatty acids (FFAs) (induced by Intralipid and heparin infusion) in members of the Sandy Lake Oji-Cree population (n = 12) and to compare the response to that in healthy age-matched nondiabetic Caucasian subjects (n = 16). The insulin secretion rate, insulin sensitivity index (S(I)), and disposition index (D(I)) (an index of insulin secretion that takes into account the ambient S(I)) were assessed in response to a 4-h graded intravenous glucose infusion followed by a 20 mmol/l 2-h hyperglycemic clamp. Total insulin secretory response to the graded glucose infusion did not change after a 48-h FFA elevation versus saline control in Caucasians and increased by approximately 30% in Oji-Cree individuals (P = 0.04 for difference between the two groups). Infusion of heparin-Intralipid reduced S(I) by approximately 40% in both groups (P = 0.002). Although D(I) was markedly reduced by heparin-Intralipid infusion in Caucasians (by approximately 40%), it was reduced by only 15% in Oji-Cree individuals (P = 0.03 for difference of response between the two groups). However, S(I) and D(I) in the Oji-Cree individuals were already much lower than in Caucasians at baseline, in keeping with the very high risk of type 2 diabetes in this population. It is concluded that Oji-Cree individuals from a community at very high risk for developing type 2 diabetes are not more susceptible to the FFA-induced desensitization of glucose stimulated insulin secretion than healthy non-Natives and, in fact, appear to be less susceptible. Whether this reflects an inherent resistance to lipotoxicity or an already-present lipotoxic effect in this population will require further study. PMID- 12765962 TI - Differential effects of diabetes on rat choroid plexus ion transporter expression. AB - Though diabetes is a disease with vascular complications, little is known about its effects on the blood-brain barrier or the blood-cerebrospinal fluid barrier (BCSFB). The BCSFB is situated at choroid plexuses located in the lateral, third, and fourth ventricles. Choroid plexuses are the primary site of cerebrospinal fluid (CSF) production and express numerous ion transporters. Previous studies have shown a perturbation of ion transport in the periphery and brain during diabetes. In this study, we investigated the effect of diabetes on ion transporters in the choroid plexuses of streptozotocin (STZ)-induced diabetic rats. Diabetes was induced in male Sprague-Dawley rats by intraperitoneal injection of STZ (60 mg/kg in citrate buffer, confirmed by glucose analysis: 601 +/- 22 mg/dl diabetic rats, 181 +/- 46 mg/dl age-matched controls); and at 28 days, rats were killed, choroid plexuses harvested, and protein extracted. Western blot analyses were carried out using antibodies for ion transporters, including Na(+)-K(+)-2Cl(-) cotransporter and the Na(+)-K(+)-ATPase alpha1 subunit. The efflux of the K(+) analog (86)Rb(+) from choroid plexus was also studied. Diabetic rats showed an increase in expression of the Na(+)-K(+)-2Cl(-) cotransporter and the Na(+)-K(+)-ATPase alpha1-subunit, as compared with age matched controls, a decrease in Na(+)-H(+) exchanger expression, and no change in Na(+)-K(+)-ATPase beta1- or beta2-subunit. The net effect of these changes was a 66% increase in (86)Rb(+) efflux from diabetic choroid plexus compared with controls. These changes in expression may affect choroid plexus ion balance and thus significantly affect CSF production in diabetic rats. PMID- 12765963 TI - A role for advanced glycation end products in diminished bone healing in type 1 diabetes. AB - The effect of type 1 diabetes on bone healing and bone formation in standardized craniotomy defects created in BALB/cByJ mice was determined. The hypothesis that advanced glycation end products (AGEs) contribute to diminished bone healing in diabetes was evaluated by assessing for the presence of the receptor for advanced glycation end products (RAGE) by immunohistochemistry in healing craniotomy defects in diabetic animals. The effect of local application of a known RAGE protein ligand, N(epsilon)-(carboxymethyl)lysine (CML)-mouse serum albumin (MSA), on craniotomy defect healing in normal animals was then assessed and compared to the effects of control MSA. Finally, evidence in support of the expression of RAGE mRNA and protein in osteoblastic cells was obtained. The results indicated that craniotomy defects in diabetic animals healed approximately 40% of the degree to which they healed in nondiabetic animals (P < 0.05). RAGE was expressed at higher levels in healing bone tissues in diabetic compared to control animals. Further studies in nondiabetic animals indicated that bone healing was reduced by 63 and 42% in lesions treated with 900 and 90 micro g CML-MSA, respectively, compared to in animals treated with MSA alone (P < 0.05). Evidence for the expression of RAGE was obtained in mouse and rat osteoblastic cultures. These results support the contribution of AGEs to diminished bone healing in type 1 diabetes, possibly mediated by RAGE. PMID- 12765964 TI - Alternative splicing of NHE-1 mediates Na-Li countertransport and associates with activity rate. AB - Sodium-lithium countertransport (SLC) is an ouabain-insensitive exchange of Na for Li found in the erythrocyte membrane of several mammalian species. Although increased SLC activity is presently the most consistent intermediate phenotype of essential hypertension and diabetic nephropathy in humans, the gene responsible for this membrane transport has not been identified. Because of functional similarities, SLC was suggested to represent an in vitro mode of operation of the Na-H exchanger (NHE). This hypothesis, however, has been long hampered by the total insensitivity of SLC to amiloride, which is an intrinsic inhibitor of the first isoform of NHE, the only NHE isoform detected in human erythrocytes. We describe here the identification in human reticulocytes and erythrocytes of an alternative splicing of NHE lacking the amiloride binding site. Transfection experiments with this spliced variant restore amiloride-insensitive, phloretin sensitive SLC activity. Expression of both regular and spliced transcripts of NHE is increased in subjects with high SLC activity. Altogether, these findings, by extending to NHE the characteristics of inheritance and predictivity previously attributed to SLC, eventually restore the candidacy of NHE isoform 1 as a gene involved in the pathogenesis of essential hypertension and diabetic nephropathy. PMID- 12765965 TI - Protein kinase C beta2-dependent phosphorylation of core 2 GlcNAc-T promotes leukocyte-endothelial cell adhesion: a mechanism underlying capillary occlusion in diabetic retinopathy. AB - Increased leukocyte-endothelial cell adhesion is a key early event in the development of retinopathy and atherogenesis in diabetic patients. We recently reported that raised activity of glycosylating enzyme [beta]1,6 acetylglucosaminyltransferase (core 2 GlcNAc-T) is responsible for increased leukocyte-endothelial cell adhesion and capillary occlusion in retinopathy. Here, we demonstrate that elevated glucose increases the activity of core 2 GlcNAc-T and adhesion of human leukocytes to retinal capillary endothelial cells, in a dose-dependent manner, through diabetes-activated serine/threonine protein kinase C beta2 (PKCbeta2)-dependent phosphorylation. This regulatory mechanism, involving phosphorylation of core 2 GlcNAc-T, is also present in polymorphonuclear leukocytes isolated from type 1 and type 2 diabetic patients. Inhibition of PKCbeta2 activation with the specific inhibitor, LY379196, attenuated serine phosphorylation of core 2 GlcNAc-T and prevented increased leukocyte-endothelial cell adhesion. Raised activity of core 2 GlcNAc-T was associated with a threefold increase in O-linked glycosylation of P-selectin glycoprotein ligand-1 on the surface of leukocytes of diabetic patients compared with age-matched control subjects. PKCbeta2-dependent phosphorylation of core 2 GlcNAc-T may thus represent a novel regulatory mechanism for activation of this key enzyme in mediating increased leukocyte-endothelial cell adhesion and capillary occlusion in diabetic retinopathy. PMID- 12765966 TI - Impact of genetic background on development of hyperinsulinemia and diabetes in insulin receptor/insulin receptor substrate-1 double heterozygous mice. AB - Type 2 diabetes is a complex disease in which genetic and environmental factors interact to produce alterations in insulin action and insulin secretion, leading to hyperglycemia. To evaluate the influence of genetic background on development of diabetes in a genetically susceptible host, we generated mice that are double heterozygous (DH) for knockout of the insulin receptor and insulin receptor substrate-1 on three genetic backgrounds (C57BL/6 [B6], 129Sv, and DBA). Although DH mice on all backgrounds showed insulin resistance, their phenotypes were dramatically different. B6 DH mice exhibited marked hyperinsulinemia and massive islet hyperplasia and developed early hyperglycemia, with 85% overtly diabetic by 6 months. By contrast, 129Sv DH mice showed mild hyperinsulinemia and minimal islet hyperplasia, and < 2% developed diabetes. DBA mice had slower development of hyperglycemia, intermediate insulin levels, and evidence of islet degeneration, with 64% developing diabetes. Thus, mice carrying the same genetic defects on different backgrounds exhibited the full spectrum of abnormalities observed in humans with type 2 diabetes, which allowed for identification of potential loci that promote development of the diabetic phenotype. PMID- 12765967 TI - Identification of interactive loci linked to insulin and leptin in mice with genetic insulin resistance. AB - Mice double heterozygous (DH) for deletion of insulin receptor and insulin receptor substrate-1 are lean, insulin resistant, and have a phenotype that strongly depends on the genetic background of the mouse. On the C57BL/6 (B6) background, DH mice develop marked hyperinsulinemia and diabetes, whereas on the 129S6 background, DH mice exhibit only mild elevations of insulin and remain free of diabetes. F2 male mice created by an intercross between these two strains exhibit a 60% incidence of diabetes and a bell-shaped distribution of insulin levels as related to glucose, reminiscent of that in humans with type 2 diabetes. These mice also exhibit a wide range of leptin levels as related to body weight. A genome-wide scan of F2 mice reveals a quantitative trait locus (QTL) related to hyperinsulinemia on chromosome 14 (D14Mit55) with a peak logarithm of odds (LOD) score of 5.6, accounting for up to 69% of this trait. A QTL with a LOD score of 3.7 related to hyperleptinemia is present on chromosome 7 at D12Mit38 (a marker previously assigned to chromosome 12) in the area of the uncoupling protein 2/3 gene cluster. This locus also interacts synergistically with D14Mit55 in development of hyperinsulinemia and with a QTL on chromosome 12 (D12Mit231) related to hyperglycemia. These data demonstrate how multiple genetic modifiers can interact and influence the development of diabetes and the phenotype of animals with genetically programmed insulin resistance and provide evidence as to the location and nature of these genes. PMID- 12765968 TI - Metabolic effects of the Gly1057Asp polymorphism in IRS-2 and interactions with obesity. AB - Insulin receptor substrate (IRS)-2 plays an important role in insulin signaling and its disruption results in diabetes in mice. In humans, the IRS-2 Gly1057Asp substitution was associated with lower risk of type 2 diabetes in lean individuals, but with a higher risk in obese individuals. To clarify the role of IRS-2 on the development of type 2 diabetes and obesity in Pima Indians, and particularly to investigate whether the effects of the Gly1057Asp polymorphism on metabolism are mediated by obesity, molecular scanning of the gene for mutations was performed and interaction of the polymorphism with obesity was tested. We identified the previously described Gly1057Asp mutation as well as a rare Asp819His mutation and four silent polymorphisms. The effect of the Gly1057Asp mutation on type 2 diabetes and obesity was tested in a large cohort of Pima Indians (n = 998). A subgroup of nondiabetic full-heritage Pima Indians (n = 233) had measurements of body composition, glucose tolerance, insulin action (M), endogenous glucose production (EGP; hyperinsulinemic clamp), acute insulin response (AIR, 25-g intravenous glucose tolerance test, n = 118 normal glucose tolerant subjects), and percutaneous fat biopsy specimens from the periumbilical region (n = 160). A total of 132 nondiabetic subjects were included in longitudinal analyses. The frequency of the Asp1057 allele was 0.6. In cross sectional analyses, subjects homozygous for the Asp1057 allele (Asp/Asp) had a higher prevalence of type 2 diabetes than heterozygote individuals and subjects homozygous for the Gly1057 allele (X/Gly, P = 0.04). There was no effect on BMI (P = 0.78) or gene-BMI interaction on the prevalence of type 2 diabetes (P = 0.57). In the nondiabetic subgroup, subjects with Asp/Asp had higher percent body fat (P = 0.01), BMI (P = 0.02), and waist circumference (P = 0.004), but there was no difference in metabolic characteristics (all P > 0.2). However, the relationship between percent body fat and fasting glucose, basal EGP, EGP during the clamp, AIR, and subcutaneous abdominal adipocyte size was significantly different in the Asp/Asp group (P for interaction = 0.02, 0.06, 0.0007, 0.08, and 0.006, respectively) compared with the X/Gly group, suggesting a more detrimental effect of Asp homozygosity on these traits with increasing percent body fat. In longitudinal analyses, among subjects in the upper tertile of change in percent body fat, those with Asp/Asp had a larger increase in fasting and postprandial glycemia and basal EGP and a larger decrease in M and AIR than subjects with X/Gly, independent of change in obesity (all P < 0.05). In conclusion, our findings suggest that the association of homozygosity for the Asp1057 allele in IRS-2 with type 2 diabetes in Pima Indians may be mediated by interaction of the polymorphism with obesity on several diabetes-related traits. PMID- 12765969 TI - Identification of a structurally distinct CD101 molecule encoded in the 950-kb Idd10 region of NOD mice. AB - Genes affecting autoimmune type 1 diabetes susceptibility in the nonobese diabetic (NOD) mouse (Idd loci) have been mapped using a congenic strain breeding strategy. In the present study, we used a combination of BAC clone contig construction, polymorphism analysis of DNA from congenic strains, and sequence mining of the human orthologous region to generate an integrated map of the Idd10 region on mouse chromosome 3. We found seven genes and one pseudogene in the 950 kb Idd10 region. Although all seven genes in the interval are Idd10 candidates, we suggest the gene encoding the EWI immunoglobulin subfamily member EWI-101 (Cd101) as the most likely Idd10 candidate because of the previously reported immune-associated properties of the human CD101 molecule. Additional support for the candidacy of Cd101 is the presence of 17 exonic single-nucleotide polymorphisms that differ between the NOD and B6 sequences, 10 causing amino acid substitutions in the predicted CD101 protein. Four of these 10 substitutions are nonconservative, 2 of which could potentially alter N-linked glycosylation. Considering our results together with those previous reports that antibodies recognizing human CD101 modulate human T-cell and dendritic cell function, there is now justification to test whether the alteration of CD101 function affects autoimmune islet destruction. PMID- 12765970 TI - Linkage and linkage disequilibrium mapping of genes influencing human obesity in chromosome region 7q22.1-7q35. AB - Linkage results suggest that the region of chromosome 7 containing the leptin gene cosegregates with extreme obesity; however, leptin coding region mutations are rare. To investigate whether the leptin flanking sequence and/or a larger 40 cM region (7q22.1-7q35) contributes to obesity, we genotyped individuals from 200 European American families segregating extreme obesity and normal weight (1,020 subjects) using 21 microsatellite markers and two single nucleotide polymorphisms (SNPs) and conducted nonparametric linkage (NPL) analyses. We also carried out transmission disequilibrium tests for 135 European American triads using 27 markers (including eight SNPs). Both quantitative (MERLIN-regress) and qualitative (GENEHUNTER and MERLIN-npl) analyses provided evidence for linkage for BMI (GENEHUNTER NPL = 2.98, 20 cM centromeric to leptin at the marker D7S692; MERLIN Z score = 3.56). Results for several other regions in 7q gave weak linkage. Transmission disequilibrium test (TDT) and quantitative TDT (and quantitative pedigree disequilibrium test) analyses suggest linkage disequilibrium near leptin and other regions of 7q. Our results suggested that there could be two or more genes in chromosome region 7q22.1-7q35 that influence obesity. A new region found by this study (D7S692-D7S523, 7q31.1) has the most consistent linkage results and could harbor obesity-related genes. PMID- 12765971 TI - Polymorphisms in the insulin-degrading enzyme gene are associated with type 2 diabetes in men from the NHLBI Framingham Heart Study. AB - Linkage studies have mapped a susceptibility gene for type 2 diabetes to the long arm of chromosome 10, where we have previously identified a quantitative trait locus that affects fasting blood glucose within the Framingham Heart Study cohort. One candidate gene in this region is the insulin-degrading enzyme (IDE), which, in the GK rat model, has been associated with nonobese type 2 diabetes. Single nucleotide polymorphisms (SNPs) were used to map a haplotype block in the 3' end of IDE, which revealed association with HbA(1c), fasting plasma glucose (FPG), and mean fasting plasma glucose (mFPG) measured over 20 years. The strongest associations were found in a sample of unrelated men. The lowest trait values were associated with a haplotype (TT, f approximately 0.32) containing the minor allele of rs2209772 and the major allele of the rs1887922 SNP (FPG P < 0.001, mFPG P < 0.003, HbA(1c) P < 0.025). Another haplotype (CC, f approximately 0.16) was associated with elevated HbA(1c) (P < 0.002) and type 2 diabetes (P < 0.001, odds ratio 1.96, 95% CI 1.28-3.00). The evidence presented supports the possibility that IDE is a susceptibility gene for diabetes in populations of European descent. PMID- 12765972 TI - Pro12Ala of the peroxisome proliferator-activated receptor-gamma2 gene is associated with lower serum insulin levels in nonobese African Americans: the Atherosclerosis Risk in Communities Study. AB - Recent research suggests that the Pro12Ala variant in peroxisome proliferator activated receptor-gamma2 (PPAR-gamma2) is associated with diabetes- and obesity related traits, and that its effects may be modified by obesity status. We characterized this variant in a population-based sample of 1,441 middle-aged African-American individuals with respect to diabetes-, obesity-, and other cardiovascular-related traits, both cross-sectionally and prospectively. The overall frequency of Ala12 was 1.9% (95% CI 1.5-2.5%), significantly lower than in Caucasian populations. Consistent with previous findings in Caucasians, African Americans with type 2 diabetes tended to be less likely to have the Pro/Ala genotype than those without (odds ratio [OR] 0.64, 95% CI 0.34-1.20); however, this OR was not statistically significant. Among nonobese individuals, the Pro/Ala genotype was associated with significantly lower ln(insulin) (P = 0.001), lower ln(HOMA-IR) (homeostasis model assessment of insulin resistance) (P = 0.002), higher fasting glucose-to-insulin ratio (P = 0.005), and lower diastolic blood pressure (P = 0.02). Among overweight individuals (BMI 25-29.9 kg/m(2)), the Pro/Ala genotype was associated with greater BMI (P = 0.02), waist to-hip ratio (P = 0.01), and waist circumference (P = 0.04). Among obese individuals, there was no association between any of the diabetes- or obesity related traits and the Pro12Ala PPAR-gamma2 variant. We conclude that among nonobese African Americans, the Pro/Ala genotype is associated with markers of greater insulin sensitivity. PMID- 12765973 TI - Prevalence of mutations in AGPAT2 among human lipodystrophies. AB - Berardinelli-Seip congenital lipodystrophy (BSCL) is a heterogeneous genetic disease characterized by near absence of adipose tissue and severe insulin resistance. We have previously identified mutations in the seipin gene in a subset of our patients' cohort. Recently, disease-causing mutations in AGPAT2 have been reported in BSCL patients. In this study, we have performed mutation screening in AGPAT2 and the related AGPAT1 in patients with BSCL or other forms of lipodystrophy who have no detectable mutation in the seipin gene. We found 38 BSCL patients from 30 families with mutations in AGPAT2. Three of the known mutations were frequently found in our families. Of the eight new alterations, six are null mutations and two are missense mutations (Glu172Lys and Ala238Gly). All the patients harboring AGPAT2 mutations presented with typical features of BSCL. We did not find mutations in patients with other forms of lipodystrophies, including the syndromes of Lawrence, Dunnigan, and Barraquer-Simons, or with type A insulin resistance. In conclusion, mutations in the seipin gene and AGPAT2 are confined to the BSCL phenotype. Because we found mutations in 92 of the 94 BSCL patients studied, the seipin gene and AGPAT2 are the two major genes involved in the etiology of BSCL. PMID- 12765974 TI - A polymorphism in the TCF7 gene, C883A, is associated with type 1 diabetes. AB - Type 1 diabetes is an autoimmune disease with a Th1 phenotype in which insulin producing beta-cells in the pancreas are destroyed. The T-cell-specific transcription factor TCF7 activates genes involved in immune regulation and is a candidate locus for genetic susceptibility to type 1 diabetes. A nonsynonymous single nucleotide polymorphism (SNP) (Pro to Thr) in the TCF7 gene, C883A, was examined in samples from 282 Caucasian multiplex type 1 diabetic families. HLA DRB1 and -DQB1 genotypes were previously determined for these samples, allowing data stratification based on HLA-associated risk. The transmission disequilibrium test showed significant overtransmission of the A allele from fathers (64.1%, P < 0.007) and nonsignificant overtransmission (57.4%, P < 0.06) of the A allele to patients who do not carry the highest-risk HLA-DR3/DR4 genotype. Elliptical sib pair analysis showed significant associations of the A allele with type 1 diabetes in paternal transmissions (P < 0.03), transmissions to male children (P < 0.04), and in the non-DR3/DR4 group (P < 0.04). These data also suggest that TCF7 C883A may affect age of disease onset. Analysis of genotype data from surrounding SNPs suggests that this TCF7 polymorphism may itself represent a risk factor for type 1 diabetes. PMID- 12765976 TI - Status of research funded by the American Diabetes Association--year 4. PMID- 12765975 TI - A quantitative trait locus on 7q31 for the changes in plasma insulin in response to exercise training: the HERITAGE Family Study. AB - Several genome-wide linkage scans have been carried out to identify quantitative trait loci for type 2 diabetes and related metabolic phenotypes. However, no previous linkage scans have focused on the response to exercise training of relevant metabolic traits. We performed a genome-wide linkage scan for baseline fasting glucose, insulin, and C-peptide and their responses to a 20-week exercise training program in nondiabetic white and black men and women from the HERITAGE Family Study. In SIBPAL linkage analyses, the maximum number of sibpairs available was 344 and 93 for baseline phenotypes and 300 and 72 for exercise training response phenotypes in whites and blacks, respectively. A total of 509 markers with an average spacing of 6.0 Mb were used. The strongest linkage was found for the changes in fasting insulin in response to exercise training with a marker in the leptin gene on 7q31 (empirical multipoint P = 0.0004) in whites. In blacks, the strongest linkage was observed for baseline fasting glucose on 12q13 q14 (empirical multipoint P = 0.0006). These regions harbor several potential candidate genes. The present findings may be important in identifying individuals at increased risk of developing type 2 diabetes and who are most likely to benefit from a physically active lifestyle. PMID- 12765977 TI - The SARS coronavirus: rapid diagnostics in the limelight. PMID- 12765978 TI - Genotype-specific influence on nitric oxide synthase gene expression, protein concentrations, and enzyme activity in cultured human endothelial cells. AB - BACKGROUND: The results of studies on the association of ecNOS polymorphisms and vascular diseases are inconsistent. To explore the nature of this interaction in the absence of confounding factors, such as smoking, we measured ecNOS mRNA, protein, and enzyme activity in cultured human umbilical vein endothelial cells (HUVECs) with and without ecNOS polymorphisms. METHODS: We identified a T(-786)- >C polymorphism in the promoter region, the intron 4 variable number of tandem repeats (VNTR), the E298A polymorphism in exon 7, and the G10-T polymorphism in intron 23 of the ecNOS gene in the DNA from 43 human umbilical cords. We measured ecNOS and GAPDH mRNA from the cultured HUVECs by reverse transcription-PCR and ecNOS protein and enzyme activity by Western blotting (as ratio to positive control band) and by determining the conversion of [(3)H]arginine to [(3)H]citrulline, respectively. RESULTS: The T(-786)-->C polymorphism showed the same allelic distribution as the intron 4 VNTR. Mean (SD) ecNOS protein from the cultured HUVECs was significantly lower in the 4a/4b genotype [0.84 (1.23); n = 9] of the intron 4 VNTR than in the 4b/4b genotype [2.14 (2.26); n = 34; P = 0.0300]. The enzyme activity was also significantly lower in the 4a/4b genotype [0.84 (0.21) pmol.min(-1).mg protein(-1); n = 9] than in the 4b/4b genotype [1.07 (0.31) pmol.min(-1).mg protein(-1); n = 34; P = 0.0197]. CONCLUSIONS: ecNOS gene expression, protein concentrations, and enzyme activity are genotype-dependent in HUVECs. The intron 4 VNTR has a consistent influence that may be mediated by the T(-786)-->C polymorphism in the promoter region. PMID- 12765979 TI - High-resolution genotyping by amplicon melting analysis using LCGreen. AB - BACKGROUND: High-resolution amplicon melting analysis was recently introduced as a closed-tube method for genotyping and mutation scanning (Gundry et al. Clin Chem 2003;49:396-406). The technique required a fluorescently labeled primer and was limited to the detection of mutations residing in the melting domain of the labeled primer. Our aim was to develop a closed-tube system for genotyping and mutation scanning that did not require labeled oligonucleotides. METHODS: We studied polymorphisms in the hydroxytryptamine receptor 2A (HTR2A) gene (T102C), beta-globin (hemoglobins S and C) gene, and cystic fibrosis (F508del, F508C, I507del) gene. PCR was performed in the presence of the double-stranded DNA dye LCGreen, and high-resolution amplicon melting curves were obtained. After fluorescence normalization, temperature adjustment, and/or difference analysis, sequence alterations were distinguished by curve shape and/or position. Heterozygous DNA was identified by the low-temperature melting of heteroduplexes not observed with other dyes commonly used in real-time PCR. RESULTS: The six common beta-globin genotypes (AA, AS, AC, SS, CC, and SC) were all distinguished in a 110-bp amplicon. The HTR2A single-nucleotide polymorphism was genotyped in a 544-bp fragment that split into two melting domains. Because melting curve acquisition required only 1-2 min, amplification and analysis were achieved in 10 20 min with rapid cycling conditions. CONCLUSIONS: High-resolution melting analysis of PCR products amplified in the presence of LCGreen can identify both heterozygous and homozygous sequence variants. The technique requires only the usual unlabeled primers and a generic double-stranded DNA dye added before PCR for amplicon genotyping, and is a promising method for mutation screening. PMID- 12765981 TI - Convenient and effective method for removing fibrinogen from serum specimens before protein electrophoresis. AB - BACKGROUND: Fibrinogen in serum specimens can be misinterpreted on protein electrophoresis as a monoclonal protein. We evaluated selective precipitation of fibrinogen with ethanol. METHODS: Pooled human plasma was mixed with absolute ethanol or saline (final concentrations of 40, 80, 100, 120, and 160 mL/L) and incubated at 4 degrees C overnight or placed in an ice bath for 15 min. After centrifugation, the supernatants and resuspended pellets were used for protein electrophoresis and quantitative measurements of protein and fibrinogen. RESULTS: The fibrinogen band was effectively eliminated from the electrophoretic pattern in the plasma samples treated with ethanol at 100 mL/L and incubated in an ice bath for 15 min without a significant change in immunoglobulin concentrations. The 100 mL/L ethanol did not noticeably change the electrophoretic pattern of monoclonal immunoglobulins. This approach allowed analysis of a sample collected from an arteriovenous shunt kept open with heparin. CONCLUSIONS: Ethanol, 100 mL/L, can selectively precipitate fibrinogen without significantly interfering with the immunoglobulins. The precipitation process can be completed in 15 min at 0-4 degrees C and can avoid the need to obtain another blood sample. PMID- 12765980 TI - Diagnostic accuracy of fecal calprotectin assay in distinguishing organic causes of chronic diarrhea from irritable bowel syndrome: a prospective study in adults and children. AB - BACKGROUND: Fecal calprotectin (FC) has been proposed as a marker of inflammatory bowel disease (IBD), but few studies have evaluated its usefulness in patients with chronic diarrhea of various causes. We evaluated the diagnostic accuracy of a FC assay in identifying "organic" causes of chronic diarrhea in consecutive adults and children. METHODS: We consecutively enrolled 70 adult patients (30 males, 40 females; median age, 35 years) and 50 children (20 males, 30 females; median age, 3.5 years) with chronic diarrhea of unknown origin. All patients underwent a complete work-up to identify the causes of chronic diarrhea. FC was measured by ELISA. RESULTS: In adult patients, FC showed 64% sensitivity and 80% specificity with 70% positive and 74% negative predictive values for organic causes. False-positive results (8 of 40 cases) were associated with the use of aspirin (3 cases) or nonsteroidal antiinflammatory drugs (1 case) and with the presence of concomitant liver cirrhosis (3 cases). False-negative results mainly included patients suffering from celiac disease (5 cases). Patients with IBD (9 cases) were identified with 100% sensitivity and 95% specificity. In pediatric patients, sensitivity was 70%, specificity was 93%, and positive and negative predictive values were 96% and 56%. False-negative results (11 of 35 cases) were associated mainly with celiac disease (6 cases) or intestinal giardiasis (2 cases). CONCLUSIONS: FC assay is an accurate marker of IBD in both children and adult patients. In adults, false negatives occur (e.g., in celiac disease) and false-positive results are seen in cirrhosis or users of nonsteroidal antiinflammatory drugs. Diagnostic accuracy is higher in children. PMID- 12765982 TI - Strategy for analysis of cardiac troponins in biological samples with a combination of affinity chromatography and mass spectrometry. AB - BACKGROUND: Cardiac troponins are modified during ischemic injury and are found as a heterogeneous mixture in blood of patients with cardiovascular diseases. We present a strategy to isolate cardiac troponins from human biological material, by use of affinity chromatography, and to provide samples ready for direct analysis by mass spectrometry. METHODS: Cardiac troponins were isolated from human left ventricular tissue by affinity chromatography. Isolated troponins were either eluted and analyzed by Western blot or enzymatically digested while bound to affinity beads. The resulting peptide mixture was subjected to mass spectrometry for protein identification and characterization. The same method was used to analyze serum from patients with acute myocardial infarction (AMI). RESULTS: Affinity chromatography with antibodies specific for one cardiac troponin subunit facilitated the isolation of the entire cardiac troponin complex from myocardial tissue. The three different proteases used for enzymatic digestion increased the total protein amino acid sequence coverage by mass spectrometry for the three cardiac troponin subunits. Combined amino acid sequence coverage for cardiac troponin I, T, and C (cTnI, cTnT, cTnC) was 54%, 48%, and 40%, respectively. To simulate matrix effects on the affinity chromatography-mass spectrometry approach, we diluted tissue homogenate in cardiac troponin-free serum. Sequence coverage in this case was 44%, 41%, and 19%, respectively. Finally, affinity chromatography-mass spectrometry analysis of AMI serum revealed the presence of cardiac troponins in a wide variety of its free and/or complexed subunits, including the binary cTnI-cTnC and cTnI-cTnC-cTnT complexes. CONCLUSIONS: Affinity chromatography-mass spectrometry allows the extraction and analysis of cardiac troponins from biological samples in their natural forms. We were, for the first time, able to directly confirm the presence of cardiac troponin complexes in human serum after AMI. This approach could assist in more personalized risk stratification as well as the search for reference materials for cardiac troponin diagnostics. PMID- 12765983 TI - Clinical and analytical performance of the liaison cardiac troponin I assay in unstable coronary artery disease, and the impact of age on the definition of reference limits. A FRISC-II substudy. AB - BACKGROUND: Measurements of cardiac troponins are currently used as the standard for the detection of myocardial injury. None of the current assays complies with the new requirements on assay imprecision as proposed by the European Society of Cardiology/American College of Cardiology. Our aim was to evaluate the clinical and analytical performance of the Liaison cardiac troponin I (cTnI) assay. METHODS: EDTA-plasma was used, and cardiac troponins were assayed with the first generation AxSYM assay, the second-generation AccuTnI assay, the third-generation Elecsys assay, and the first-generation Liaison assay. RESULTS: In a 6-day imprecision study, the Liaison cTnI assay had mean CV < or =10% at 0.027 microg/L and < or =20% at 0.015 microg/L. The 99th percentile of the upper reference limit (URL) of a reference population was 0.041 microg/L (age range, 41-76 years). Individuals <60 years had a significantly (P = 0.001) lower 99th percentile, 0.022 microg/L. The FRISC-II study participants with cTnI > or =0.041 microg/L had a poorer outcome relating to death/acute myocardial infarction than those with cTnI <0.041 microg/L (P <0.001). Treatment with low-molecular-weight heparin (dalteparin) or an invasive strategy reduced cardiac events only in patients with concentrations >0.041 microg/L (P = 0.002 and 0.02, respectively). Comparison with the AccuTnI assay showed that a large cohort of the patients with poor prognosis was identified by the AccuTnI assay but not by the Liaison cTnI assay. CONCLUSION: The Liaison cTnI assay is a sensitive assay with a CV < or =10% at the 99th percentile URL. The ability to detect age-related differences among apparently healthy individuals is unique among today's commercial assays. The results indicate that different assays seem to identify different patient cohorts for cardiac risk in the lower range of cTnI concentrations. PMID- 12765984 TI - Determination of non-alpha1-antichymotrypsin-complexed prostate-specific antigen as an indirect measurement of free prostate-specific antigen: analytical performance and diagnostic accuracy. AB - BACKGROUND: A new assay measures prostate-specific antigen (PSA) not complexed to alpha(1)-antichymotrypsin (nACT-PSA) after removing PSA complexed to ACT by use of anti-ACT antibodies. We evaluated nACT-PSA and its ratio to total PSA (tPSA) as alternatives to free PSA (fPSA) and its ratio to tPSA in differentiating prostate cancer (PCa) and benign prostatic hyperplasia (BPH) in patients with tPSA of 2-20 micro g/L. METHODS: PSA in serum of 183 untreated patients with PCa and 132 patients with BPH was measured retrospectively on the chemiluminescence immunoassay analyzer LIAISON(R) (Byk-Sangtec Diagnostica) with the LIAISON tPSA and LIAISON fPSA assays. The nACT-PSA fraction was determined with a prototype assay measuring the residual PSA after precipitation of ACT-PSA with an ACT precipitating reagent. RESULTS: nACT-PSA was higher than fPSA in samples with fPSA concentrations <1 microg/L but lower in samples with >1 microg/L fPSA. The median ratios of fPSA/tPSA and of nACT-PSA/tPSA were significantly different between patients with BPH and PCa (19.4% vs 12.2% and 17.4% vs 13.0%, respectively). Within the tPSA ranges tested (2-20, 2-10, and 4-10 microg/L), areas under the ROC curves for the fPSA/tPSA ratios were significantly larger than those for nACT-PSA/tPSA. In the tPSA ranges <10 microg/L, the areas under the ROC curves for fPSA/tPSA were significantly larger than those for tPSA, whereas the areas for nACT-PSA/tPSA were not. At decision limits for 95% sensitivity and specificity, both ratios significantly increased specificity and sensitivity, respectively, compared with tPSA, but the fPSA/tPSA ratio showed higher values. CONCLUSIONS: nACT-PSA and its ratio to tPSA provide lower diagnostic sensitivity and specificity than fPSA/tPSA. The fPSA/tPSA ratio represents the state-of-the-art method for differentiating between PCa and BPH. PMID- 12765985 TI - Plasma total cysteine, mortality, and cardiovascular disease hospitalizations: the Hordaland Homocysteine Study. AB - BACKGROUND: We have previously reported a positive association between tHcy and mortality and cardiovascular disease (CVD) hospitalizations in the Hordaland Homocysteine Study cohort. Using the same data set, we assessed the relationship between plasma total cysteine (tCys) and mortality from all causes and from cardiovascular and noncardiovascular conditions, and the association between tCys and the risk of hospitalizations from CVD. METHODS: We measured plasma tCys in blood samples from 12,595 men and women 40-42 years of age and from 4766 men and women 65-67 years of age, collected as part of the Hordaland Homocysteine Study in the year 1992-1993. Follow-up data on mortality were collected through 1999. Data on CVD hospitalizations were collected from hospital records up to May 31, 1998. RESULTS: After a follow-up time of 6.6-7.6 years, there were a total of 610 deaths, of which 243 were cardiovascular deaths and 367 were noncardiovascular deaths. There was no association between tCys and all-cause, cardiovascular, or noncardiovascular mortality. When we used tCys values <247.6 micromol/L (lowest quartile) as the reference category, the adjusted mortality ratio (MR) for all cause mortality at tCys concentrations of 247.6-270.79, 270.8-295.79, and > or =295.8 micromol/L (highest quartile) were 1.0, 0.9, and 1.0, respectively. The adjusted MRs for cardiovascular mortality were 1.0, 1.1, and 1.1, respectively. There were no associations between tCys and 1275 CVD hospitalizations, except that tCys was significantly associated with hospitalizations from coronary artery bypass grafting. CONCLUSION: Plasma tCys is not associated with mortality or CVD hospitalizations. PMID- 12765986 TI - Detection of recombinant human erythropoietin in urine by isoelectric focusing. AB - BACKGROUND: Doping with erythropoietic proteins such as recombinant human erythropoietin (rHuEPO) and darbepoetin alfa is a serious issue in sport. There is little information on the time course of detection of rHuEPO in urine and on methods to evaluate electrophoresis-based data. METHODS: We used a recently described isoelectric focusing method for detecting rHuEPO and endogenous EPO in urine obtained from individuals treated with placebo or epoetin alfa. The latter was administered subcutaneously at 50 IU/kg on days 0, 2, 4, 7, 9, 11, 14, 16, and 18. Blood and urine samples were collected during the morning of study days 3, 0, 2, 4, 7, 9, 11, 14, 16, and 18 and on days 2, 3, 4, and 7 postadministration. We developed visual and numerical (two-band ratio) techniques to evaluate the electropherograms for the presence of rHuEPO. RESULTS: Compared with the placebo group, the epoetin alfa-treated group responded with increases in hematocrit, reticulocytes, macrocytes, serum EPO, and serum soluble transferrin receptor. The electropherograms showed that the pattern of bands arising from urinary rHuEPO is different from that of endogenous urinary EPO. Both the two-band ratio and the visual technique detected rHuEPO in all 14 epoetin alfa-treated individuals 3 days after the last dose. On the 7th day after the last dose, both techniques detected rHuEPO in approximately one-half of the participants. rHuEPO was not detected in the placebo-treated individuals. CONCLUSIONS: The isoelectric focusing method detects rHuEPO in most urine samples collected 3 days after nine doses of epoetin alfa. The numerical two-band ratio was equivalent to a visual method for detecting rHuEPO in urine. PMID- 12765987 TI - Time-resolved fluorometric assay for detection of autoantibodies to glutamic acid decarboxylase (GAD65). AB - BACKGROUND: Type 1 diabetes mellitus results from destruction of the pancreatic insulin-producing beta cells by a chronic autoimmune process. Methods are needed for the detection of circulating autoantibodies to glutamic acid decarboxylase (GAD65), a major marker of this process. METHODS: Streptavidin-coated microtiter plates were incubated with biotinylated GAD65, and after incubation with serum samples from patients with type 1 diabetes mellitus and control individuals, europium-labeled GAD65 was added. After washing steps, the delayed fluorescence was measured in duplicate in a fluorometer. Samples collected from 100 patients with newly diagnosed type 1 diabetes mellitus and 100 healthy controls were measured by the new assay and by a radiobinding assay. RESULTS: The detection limit of the new assay was 1.49 WHO units/mL, the calibration curve was linear to 4140 WHO units/mL, and no hook effect was observed up to 41,400 WHO units/mL. The intraassay CV was 2.1-6.3% over the calibration range. For patient serum samples, the intraassay, interassay, and total CVs were 5.4-7.0%, 9.8-13%, and 12-14%, respectively. Compared with conventional radioimmunologic methods, the analytical range was broader and the analysis time required to perform the measurements was shorter. At a cutoff with 99% specificity, the new assay and the radiobinding assay were positive in 71 and 67 patients, respectively. CONCLUSIONS: The new assay provides a rapid and sensitive nonradioactive method applicable for large scale screening for beta-cell autoimmunity. It has a broad linear analytical range, is easy to perform and automate, and has sensitivity and specificity comparable to those for the conventional radioisotope assay. PMID- 12765988 TI - Detection of autoantibodies to protein tyrosine phosphatase-like protein IA-2 with a novel time-resolved fluorimetric assay. AB - BACKGROUND: Circulating autoantibodies to pancreatic glutamic acid decarboxylase (GAD65; the 65-kDa isoform of glutamic acid decarboxylase), protein tyrosine phosphatase-like protein IA-2, and insulin can be used as predictive markers of type 1 diabetes. We developed a novel assay for the detection of IA-2 autoantibodies (IA-2As) in serum based on time-resolved fluorimetry, hypothesizing that this kind of assay could provide several advantages over methods described to date, including radiobinding assays (RBAs) and ELISAs. METHODS: The intracellular part of IA-2 (IA-2ic) was biotinylated and bound to streptavidin-coated 96-well plates by simultaneous incubation with serum samples and glutathione S-transferase (GST)-IA-2ic fusion protein. GST-IA-2ic captured by autoantibodies in the serum was detected with europium-labeled anti-GST antibody, and the signal was measured in a time-resolved fluorimeter. A serum sample panel from 100 patients with newly diagnosed type 1 diabetes and 100 unaffected controls was analyzed with the new assay and a conventional RBA. RESULTS: Among the 100 serum samples from patients with type 1 diabetes, the time-resolved fluorimetric assay identified 74 IA-2A-containing sera, whereas the RBA detected 80 IA-2A-positive samples. Five of the six samples positive in the RBA but not detected by the time-resolved fluorimetric assay were only weakly positive in the RBA. The performance time of the time-resolved fluorimetric assay was 2.5 h compared with 10-12 h required by the RBA. CONCLUSIONS: The time-resolved fluorimetric assay provides a simple, nonradioactive analysis method for the detection of IA-2As with a specificity and a sensitivity comparable to the RBA method. This assay allows substantial reduction in performance time compared with the conventional RBA. PMID- 12765989 TI - Monitoring blood glucose changes in cutaneous tissue by temperature-modulated localized reflectance measurements. AB - BACKGROUND: Most proposed noninvasive methods for glucose measurements do not consider the physiologic response of the body to changes in glucose concentration. Rather than consider the body as an inert matrix for the purpose of glucose measurement, we exploited the possibility that noninvasive measurements of glucose can be approached by investigating their effects on the skin's thermo-optical response. METHODS: Glucose concentrations in humans were correlated with temperature-modulated localized reflectance signals at wavelengths between 590 and 935 nm, which do not correspond to any near-infrared glucose absorption wavelengths. Optical signal was collected while skin temperature was modulated between 22 and 38 degrees C over 2 h to generate a periodic set of cutaneous vasoconstricting and vasodilating events, as well as a periodic change in skin light scattering. The method was tested in a series of modified meal tolerance tests involving carbohydrate-rich meals and no-meal or high-protein/no-carbohydrate meals. RESULTS: The optical data correlated with glucose values. Changes in glucose concentrations resulting from a carbohydrate rich meal were predicted with a model based on a carbohydrate-meal calibration run. For diabetic individuals, glucose concentrations were predicted with a standard error of prediction <1.5 mmol/L and a prediction correlation coefficient 0.73 in 80% of the cases. There were run-to-run differences in predicted glucose concentrations. Non-carbohydrate meals showed a high degree of scatter when predicted by a carbohydrate meal calibration model. CONCLUSIONS: Blood glucose concentrations alter thermally modulated optical signals, presumably through physiologic and physical effects. Temperature changes drive cutaneous vascular and refractive index responses in a way that mimics the effect of changes in glucose concentration. Run-to-run differences are attributable to site-to-site structural differences. PMID- 12765990 TI - Preanalytical factors that influence the Abbott TDx Fetal Lung Maturity II assay. AB - BACKGROUND: The TDx Fetal Lung Maturity II (FLM II) assay uses amniotic fluid to assess lung maturity of the unborn infant. We investigated common preanalytical factors that influence FLM results, including centrifugation, sample storage, and contamination by whole blood. METHODS: We tested 18 specimens after centrifugation and after resuspension by vortex-mixing. We also analyzed 23 specimens stored at -20 degrees C for up to 448 days and then thawed (duplicate measurements), 20 specimens stored at 4 degrees C, and 24 specimens stored at room temperature. In addition, we evaluated the effects of whole blood diluted into 19 different specimens. RESULTS: Centrifugation significantly decreased FLM II results from baseline (P <0.0001), and resuspension returned results to baseline values (P = 0.286). Storage at -20 degrees C produced highly variable results that demonstrated a nonsignificant negative trend associated with storage time. Specimens were stable for 24 h when stored at 4 degrees C and 16 h at room temperature. Blood contamination produced significantly positive differences in results only in specimens with baseline values < or =39 mg/g with a 5.8 mg/g increase in FLM II for every 0.1 x 10(12)/L increase in the erythrocyte count (slope = 58.4). CONCLUSIONS: Resuspension of centrifuged specimens produces clinically valid FLM II results. Results from specimens stored at -20 degrees C can be highly variable and decrease over time. Results from specimens stored at 4 degrees C and at room temperature are stable for 24 and 16 h, respectively. Blood contamination up to 0.03 x 10(12) erythrocytes/L is acceptable for FLM II analysis. PMID- 12765991 TI - Chemiluminescence assay improves specificity of hepatitis C antibody detection. AB - BACKGROUND: Antibodies to hepatitis C virus (anti-HCV) have typically been detected by enzyme immunoassay (EIA). A chemiluminescence assay (CA) for anti-HCV is now commercially available. METHODS: We compared the positive rate for a CA in a HCV screening program for veterans with historical rates obtained with EIA. We also compared results in 2824 samples tested by both methods and assessed the significance of low signal-to-cutoff (S/C) ratios. RESULTS: The frequency of CA positive results was significantly lower than with EIA (12.6% vs 16.0%; P <0.0001). The frequency of low S/C ratios was also significantly lower with CA (11.5% vs 20.0%; P <0.0001). Among low-positive values, samples positive by CA were significantly less likely to be recombinant immunoblot assay (RIBA)-negative (64% vs 84%; P <0.0005). In parallel testing, results for 111 samples (3.9%) were discrepant between the two assays; all but 6 had low S/C ratios, and confirmatory testing was performed on all but 8 samples. Of 56 EIA-positive, CA-negative samples tested by RIBA, only 1 was positive. Of 24 CA-positive, EIA-negative samples, 62% were RIBA-negative. Using a negative RIBA result as an indication of false-positive anti-HCV results, the positive predictive value of EIA was 93% compared with 98% with CA. HCV RNA was positive in 90% of samples high-positive by both CA and EIA. Only 2 of 30 (7%) low-positive CA samples were RNA-positive. CONCLUSIONS: CA produces fewer false-positive and fewer low-positive results that require confirmatory RIBA testing. The S/C ratio remains useful for characterizing positive results. PMID- 12765992 TI - Hydroxytyrosol disposition in humans. AB - BACKGROUND: Animal and in vitro studies suggest that phenolic compounds in virgin olive oil are effective antioxidants. In animal and in vitro studies, hydroxytyrosol and its metabolites have been shown to be strong antioxidants. One of the prerequisites to assess their in vivo physiologic significance is to determine their presence in human plasma. METHODS: We developed an analytical method for both hydroxytyrosol and 3-O-methyl-hydroxytyrosol in plasma. The administered dose of phenolic compounds was estimated from methanolic extracts of virgin olive oil after subjecting them to different hydrolytic treatments. Plasma and urine samples were collected from 0 to 12 h before and after 25 mL of virgin olive oil intake, a dose close to that used as daily intake in Mediterranean countries. Samples were analyzed by capillary gas chromatography-mass spectrometry before and after being subjected to acidic and enzymatic hydrolytic treatments. RESULTS: Calibration curves were linear (r >0.99). Analytical recoveries were 42-60%. Limits of quantification were <1.5 mg/L. Plasma hydroxytyrosol and 3-O-methyl-hydroxytyrosol increased as a response to virgin olive oil administration, reaching maximum concentrations at 32 and 53 min, respectively (P <0.001 for quadratic trend). The estimated hydroxytyrosol elimination half-life was 2.43 h. Free forms of these phenolic compounds were not detected in plasma samples. CONCLUSIONS: The proposed analytical method permits quantification of hydroxytyrosol and 3-O-methyl-hydroxytyrosol in plasma after real-life doses of virgin olive oil. From our results, approximately 98% of hydroxytyrosol appears to be present in plasma and urine in conjugated forms, mainly glucuronoconjugates, suggesting extensive first-pass intestinal/hepatic metabolism of the ingested hydroxytyrosol. PMID- 12765993 TI - Rapid diagnosis of a coronavirus associated with severe acute respiratory syndrome (SARS). PMID- 12765994 TI - Automated RNA extraction by MagNA Pure followed by rapid quantification of cytokine and chemokine gene expression with use of fluorescence resonance energy transfer. PMID- 12765995 TI - Stability of NT-proBNP in serum specimens collected in Becton Dickinson Vacutainer (SST) tubes. PMID- 12765996 TI - Physical characteristics of six new thermocyclers. PMID- 12765997 TI - Reproducibility of blood markers of oxidative status and endothelial function in healthy individuals. PMID- 12765998 TI - Liquid chromatography-tandem mass spectrometry analysis of urinary free cortisol. PMID- 12765999 TI - Pediatric concentrations of S100B protein in blood: age- and sex-related changes. PMID- 12766000 TI - Quantitative spectrophotometric microplate assay for angiotensin-converting enzyme in cerebrospinal fluid. PMID- 12766001 TI - Improved indicators for assessing the reliability of detection and quantification by kinetic PCR. PMID- 12766002 TI - Comparison of the Biomedica NT-proBNP enzyme immunoassay and the Roche NT-proBNP chemiluminescence immunoassay: implications for the prediction of symptomatic and asymptomatic structural heart disease. PMID- 12766003 TI - Extraction of RNA from dried blood on filter papers after long-term storage. PMID- 12766004 TI - Fetal expressed gene analysis in maternal blood: a new tool for noninvasive study of the fetus. PMID- 12766005 TI - Functional hyperhomocysteinemia in healthy vegetarians: no association with advanced glycation end products, markers of protein oxidation, or lipid peroxidation after correction with vitamin B(12). PMID- 12766006 TI - Bilirubin in amniotic fluid does not interfere with the Abbott TDx FLM II assay. PMID- 12766007 TI - Automated latex photometric immunoassay for total plasminogen activator inhibitor 1 in plasma. PMID- 12766008 TI - Detection of mutations in the hepatitis B virus polymerase gene. PMID- 12766009 TI - Standardization of C-peptide measurements in urine by method comparison with isotope-dilution mass spectrometry. PMID- 12766010 TI - Lamellar body count in amniotic fluid: a comparative study of four different hematology analyzers. PMID- 12766011 TI - S100B protein concentrations in amniotic fluid are higher in monoamniotic than in diamniotic twins and singleton pregnancies. PMID- 12766012 TI - Cytometric bead array to measure six cytokines in twenty-five microliters of serum. PMID- 12766013 TI - The serum growth hormone (GH) response to provocative tests is dependent on type of assay in autosomal dominant isolated GH deficiency because of an ARG(183)HIS (R183H) GH-I gene mutation. PMID- 12766014 TI - Fasting and post-methionine loading concentrations of homocysteine, vitamin B2, and vitamin B6 in patients on antiepileptic drugs. PMID- 12766015 TI - CYP2D6 poor metabolizer status can be ruled out by a single genotyping assay for the -1584G promoter polymorphism. PMID- 12766016 TI - Reevaluation of formulas for predicting creatinine clearance in adults and children, using compensated creatinine methods. PMID- 12766018 TI - Utilization, reliability, and clinical impact of point-of-care testing during critical care transport: six years of experience. PMID- 12766017 TI - Multiple lipoprotein abnormalities associated with insulin resistance in healthy volunteers are identified by the vertical auto profile-II methodology. PMID- 12766019 TI - Use of inductively coupled plasma mass spectrometry to measure urinary iodine in NHANES 2000: comparison with previous method. PMID- 12766020 TI - Asialotransferrin--an alternative to carbohydrate-deficient transferrin? PMID- 12766021 TI - Modification of the colorimetric assay for serum unsaturated iron-binding capacity. PMID- 12766022 TI - Increased carbohydrate-deficient transferrin of unknown etiology in a 15-year-old male patient with autoimmune hepatitis type 1. PMID- 12766023 TI - Influence of centrifugation temperature on the plasma total homocysteine concentration. PMID- 12766024 TI - Changes in concentration of DNA in serum and plasma during storage of blood samples. PMID- 12766025 TI - Automated enzymatic assay for homocysteine. PMID- 12766031 TI - In vivo imaging of embryonic development in the mouse eye by ultrasound biomicroscopy. AB - PURPOSE: New imaging tools now provide an unprecedented opportunity to visualize anatomic and functional development of the mouse eye. In this study, normal embryonic development of the mouse eye was studied by ultrasound biomicroscopy (UBM), with a focus on the formation of the retina, lens, and cornea. METHODS: The growth of 65 embryonic eyes from timed-pregnant CD-1 mice was examined at various stages of development between embryonic day (E)11.5 and E18.5, using 40 MHz UBM. RESULTS: The morphogenesis of ocular tissues including the lens, retina, and orbit were revealed from the earliest stages of development. The major axis of the CD-1 lens grows at a rate of 68 micro m/d, whereas that of the globe grows at a rate of 122 microm/d, with a concomitant exponential increase in volume. CONCLUSIONS: UBM allows noninvasive assessment of ocular morphogenesis in vivo and can be used to calculate relative growth rates of ocular structures. PMID- 12766032 TI - Age-dependent iris abnormalities in collagen XVIII/endostatin deficient mice with similarities to human pigment dispersion syndrome. AB - PURPOSE: Collagen XVIII is expressed in ocular basement membranes (BMs) and inactivating mutations cause Knobloch syndrome, with several ocular abnormalities. In this study we investigated ocular structures in collagen XVIII/endostatin (Col18a1(-/-))-deficient mice to elucidate the role of this extracellular matrix component in the eye. METHODS: Eyes of Col18a1(-/-) and control mice were examined by light and transmission electron microscopy, laser scanning ophthalmoscopy, and fluorescence angiography. Immunohistochemical analysis of neuronal, epithelial, and immune cells in the eye was performed with antibodies against established cell markers. RESULTS: Col18a1(-/-) mice showed a disruption of the posterior iris pigment epithelial (IPE) cell layer with release of melanin granules. The BM of the posterior IPE was attached to the lens and the nonpigmented epithelium of the ciliary body, which was flattened in mutant mice. In aged mutant mice a severe thickening of the stromal iris BM zone was found, and pigmented cells migrated out of the iris and covered the retina along the inner limiting membrane (ILM), sometimes penetrating into the retina. These cells resembled iris clump cells, and immunohistochemistry demonstrated that they were macrophage-like cells. Furthermore, morphologically abnormal retinal vasculature was seen by fluorescence angiography. CONCLUSIONS: The abnormalities in the iris and ciliary body of Col18a1(-/-) mice demonstrate an important role of collagen XVIII for the function of ocular BMs. The absence of this collagen alters the properties of BMs and leads to severe defects in the iris, showing striking similarities to human pigment dispersion syndrome. In addition, loss of collagen XVIII creates changes that allow clump cells to migrate out of the iris. These cells have not been well characterized previously. In the current study we showed that they are macrophage-like cells and are able to penetrate the ILM in mutant mice. The disease mechanism of human pigment dispersion syndrome is not well understood, but Col18a1(-/-) mice may serve as a model and demonstrate the potential importance of alterations in extracellular matrix components in this disease. PMID- 12766033 TI - Ocular measurements throughout the adult life span of rhesus monkeys. AB - PURPOSE: To examine the relationship of ocular components to refraction throughout the adult life span of the rhesus monkey (Macaca mulatta). METHODS: Cycloplegic retinoscopy, A-scan ultrasonography, slit lamp examination, indirect ophthalmoscopy, and keratometry were performed in a cross-sectional study of 111 monkeys, aged 5 to 31 years. Lens thickness and anterior and vitreous chamber depths were measured from the echograms. The intercorrelations of these variables were analyzed, as well as their association with age and sex. RESULTS: In monkeys aged 5 to 15 years, the mean refractive value of +1.5 D with an SD of 1.7 D was maintained near the previously established developmental asymptote of +2 D. In monkeys older than 15 years, there was greater interindividual variation (SD = 4.5 D), including extreme myopia and hyperopia. The cornea became steeper with age. The axial length of the eyes increased up to 12 years of age and began to shorten after 20 years. Changes also occurred in the other individual components that constitute eye length. These age-related changes were decreased vitreous chamber depth, decreased anterior chamber depth, and increased lens thickness. In general, males had longer eyes than females. The eyes of old monkeys were more likely to exhibit cataract and drusen, but age-related changes in focal atrophy of the retinal pigment epithelium did not achieve statistical significance. CONCLUSIONS: The components of the monkey eye change with age in a pattern similar to that reported in humans. Age-related changes in individual ocular components that could be detrimental to refraction appear to be compensated for by changes in other components. PMID- 12766034 TI - Nucleolar diameter and microvascular factors as independent predictors of mortality from malignant melanoma of the choroid and ciliary body. AB - PURPOSE: To determine whether nucleolar diameter and microvascular factors are independent predictors of mortality in malignant uveal melanoma of the choroid and ciliary body. METHOD: A population-based retrospective cohort study was conducted of melanoma-specific and all-cause mortality in 167 consecutive patients who had an eye enucleated because of choroidal and ciliary body melanoma from 1972 through 1981. The largest nucleoli were measured from digital photographs of silver-stained tumors along a central 5-mm-wide linear field parallel to the base of the tumor. The mean of the 10 largest nucleoli (MLN) was calculated. Microvascular loops and networks and microvascular density (MVD) were assessed. Kaplan-Meier and Cox regression analyses were performed. Associations between MLN and other variables were determined. RESULTS: The MLN could be determined in 126 (75%) melanomas. It ranged from 2.60 to 6.18 microm (median, 4.05). The association of large MLN with the presence of epithelioid cells (P = 0.017) and high MVD (P = 0.0053) was statistically significant. MLN was not significantly associated with tumor diameter and microvascular loops and networks. The 10-year melanoma-specific survival decreased with MLN (0.74, 0.60, and 0.42, arranged in tertiles; P = 0.0060), presence of loops and networks (P = 0.0001), and increasing MVD (P = 0.0001). By Cox regression, MLN was an independent predictor of survival, when adjusting in turn for presence of epithelioid cells, loops and networks, and MVD. In multivariate models with MVD, the independent prognostic information carried by MLN decreased, but the model as a whole was a better predictor of survival. The magnitude of this effect depended on whether MLN was modeled as a continuous or categorical variable. CONCLUSIONS: In this population-based data set, MLN and microvascular loops and networks were unrelated, independent predictors of survival. MLN and MVD were found to be partially interrelated. Multivariate models that included MVD in addition to MLN fitted better with observed melanoma-specific survival than models that excluded MVD. PMID- 12766036 TI - Expression of the lipogenic enzyme fatty acid synthase (FAS) in retinoblastoma and its correlation with tumor aggressiveness. AB - PURPOSE: Fatty acid synthase (FAS) performs the anabolic conversion of dietary carbohydrate or protein to fatty acids. Many common human cancers express high levels of FAS, and its differential expression between normal and neoplastic tissues has led to the consideration of FAS as a target for anticancer therapy. To investigate the potential of targeting FAS in the treatment of retinoblastoma, we first determined whether FAS was activated in this human tumor. Moreover, correlation of FAS expression with tumor aggressiveness was determined. METHODS: FAS reactivity was evaluated by immunohistochemistry in 66 retinoblastoma specimens from 65 patients. Degree of tumor differentiation, choroid invasion, optic nerve infiltration, mitotic rate, and necrosis extension were estimated. FAS expression was correlated with all these tumor characteristics by means of parametric and nonparametric statistical analyses. RESULTS: Eighty-two percent of tumors were FAS positive. Stronger FAS expression correlated with more advanced choroid (P < 0.001) and optic nerve (P = 0.016) invasion, high mitotic index (P < 0.001), and less differentiated histology (P = 0.047). Correlation with extension of necrosis was not statistically significant. Unaffected retina was negative. CONCLUSIONS: The data suggest that expression of FAS and fatty acid synthesis support an essential functional aspect of retinoblastoma cells, perhaps cell growth or survival. FAS activation may serve as a novel target for systemic and local antineoplastic therapy and, because it increases with tumor aggressiveness, its inhibition could represent an alternative treatment strategy in advanced and resistant retinoblastomas. PMID- 12766035 TI - Depsipeptide (FR901228) inhibits proliferation and induces apoptosis in primary and metastatic human uveal melanoma cell lines. AB - PURPOSE: Uveal melanoma (UM) is the most common primary malignant ocular tumor in adults. No effective chemotherapy regimens are available for either intraocular or metastatic uveal melanoma. Therefore, the ability of the histone deacetylase inhibitors (HDACIs), depsipeptide, sodium butyrate (NaB) and trichostatin A (TSA), to induce apoptosis and inhibit cell growth of UM cell lines in vitro was examined. METHODS: Three primary and two metastatic UM cell lines were treated in vitro with different concentrations of histone deacetylase inhibitors (HDACIs). Cell proliferation was studied in 24-well plates. Induction of apoptosis was studied by flow cytometry. Changes in gene expression of Fas/FasL, p21(Waf/Cip1), and p27(Kip1) were studied by RT-PCR. Western blot analysis was used to study histone acetylation, Fas/FasL, p21(Waf/Cip1), p27(Kip1) and caspase-3 protein levels. Real-time PCR was used to study changes in bcl-2/bax gene expression. RESULTS: A dose-dependent increase in histone acetylation was observed in all cell lines. This corresponded to significant inhibition of cell growth and induction of apoptosis in all melanoma cell lines in a concentration-dependent manner. Western blot analysis revealed dose-dependent increases in the amount of caspase-3, Fas/FasL, p21(Waf/Cip1), and p27(Kip1) proteins. However, no changes in bcl-2/bax gene expression were detected by real-time PCR. CONCLUSIONS: HDACIs are potent inhibitors of primary and metastatic UM cell growth in vitro. The apoptosis is probably mediated through the Fas/FasL signaling pathway, whereas bcl-2 appears not to be involved. These data support further clinical evaluation of depsipeptide and other HDACIs in patients with primary and metastatic UM. PMID- 12766037 TI - Microphthalmia resulting from MSX2-induced apoptosis in the optic vesicle. AB - PURPOSE: Microphthalmia is a relatively common ocular malformation. Molecular mechanisms that lead to this dire condition are largely unknown. Msx genes have been shown to be expressed in the developing eye. In the Msx1;Msx2, double mutant mouse, eye development arrests early in embryogenesis. To investigate possible functions of Msx2 in early ocular development, we created transgenic animals that overexpress Msx2. METHODS: Msx2 transgenic embryos and nontransgenic littermates were examined histopathologically. The effect of Msx2 overexpression on retinal cell proliferation was assayed by bromodeoxyuridine (BrdU) incorporation and immunohistochemical staining. Apoptosis was determined by TUNEL labeling. Expression of retina and retinal pigmented epithelium (RPE)-specific genes was investigated by performing in situ hybridization or immunohistochemical staining. RESULTS: Forced expression of the Msx2 gene resulted in optic nerve aplasia and microphthalmia in all transgenic animals. In developing retinas of Msx2 transgenic animals, proliferation was significantly reduced and increased numbers of retinal cells underwent apoptosis. Marker analysis showed suppression of Bmp4 and induction of Bmp7 gene expression in the optic vesicle. Ectopic concurrent expression of the RPE cell markers Cx43 and Trp-2 in the neural retinal layer suggests cell fate respecification. CONCLUSION: These results indicate that forced expression of Msx2 perturbs BMP signaling in the developing eye and is accompanied by an increase in retinal cell death and a reduction in cell proliferation. Thus, deregulated Msx2 gene expression may be a plausible genetic mechanism by which the autosomal dominant form of congenital microphthalmia may arise. PMID- 12766038 TI - RPGR isoforms in photoreceptor connecting cilia and the transitional zone of motile cilia. AB - PURPOSE: The retinitis pigmentosa guanosine triphosphatase (GTPase) regulator (RPGR) is essential for photoreceptor survival. There is as yet no consensus concerning the subcellular localization of RPGR. This study was undertaken as a comprehensive effort to resolve current controversies. METHODS: RPGR in mice and other mammalian species was examined by immunofluorescence. RPGR variants were distinguished by using isoform-specific antibodies. Different tissue processing procedures were evaluated. Immunoblot analysis of serial cross-sections of photoreceptors was performed as a complementary approach to subcellular localization. RESULTS: RPGR was found in the connecting cilia of rods and cones with no evidence for species-dependent variation. RPGR ORF15 was the predominant variant in photoreceptor connecting cilia whereas constitutive RPGR (default) was the sole variant in the transitional zone of motile cilia in airway epithelia. Removal of soluble materials in the interphotoreceptor matrix facilitated detection of RPGR in the connecting cilia in photoreceptors. CONCLUSIONS: RPGR localizes in photoreceptor connecting cilia and in a homologous structure, the transitional zone of motile cilia. These data are important for understanding the multitude of clinical manifestations associated with mutations in RPGR. Interphotoreceptor matrix surrounding the connecting cilia is a key variable for in situ detection of a protein in the connecting cilia. PMID- 12766039 TI - Ocular and scleral alterations in gene-targeted lumican-fibromodulin double-null mice. AB - PURPOSE: To elucidate the role of leucine-rich proteoglycans lumican and fibromodulin in the sclera. METHODS: Lumican- and fibromodulin-null heterozygous mice were intercrossed to obtain wild-type (Lum(+/+)Fmod(+/+)), lumican-null (Lum(-/-)Fmod(+/+)), fibromodulin-null (Lum(+/+)Fmod(-/-)), and double-null (Lum( /-)Fmod(-/-)) littermates. Axial length was measured on enucleated whole eyes, and ocular structural changes were examined by histology. The morphology of collagen fibrils in the sclera was examined by transmission electron microscopy (TEM). RESULTS: Compared with the ocular axial length in wild type mice, the axial length was increased by 10% in Lum(-/-)Fmod(-/-) (P = 0.02) mice. Retinal detachment was frequent in the double-null and rare in the lumican-null animals. Compared with the wild-type sclera, the sclera in all null mutants was significantly thinner with fewer lamellae (P < 0.05). The double-null sclera contained abnormally large-diameter (120-160 nm) and small-diameter (30-60 nm) collagen fibrils, whereas the fibromodulin-null sclera was enriched for the small diameter fibrils. The collagen fibril diameter distribution in the lumican-null sclera was similar to that of the wild-type. CONCLUSIONS: An increase in small diameter fibrils in the fibromodulin-null sclera suggests a key role for fibromodulin in the maturation and assembly of scleral collagen fibrils. That fibril diameter distribution in the lumican-null sclera was comparable to that in the wild type, but severely disrupted in the double null, suggests a role for lumican that is crucial in the absence of fibromodulin. The eyes of Lum(-/-)Fmod( /-) mice show certain features of high myopia: increased axial length, thin sclera, and retinal detachment. Mutations or altered expression of these proteoglycans may contribute to myopia in humans. PMID- 12766041 TI - A strong and highly significant QTL on chromosome 6 that protects the mouse from age-related retinal degeneration. AB - PURPOSE: BALB/cByJ (C) albino mice have significantly more retinal degeneration as they age than C57BL/6J-c(2J) (B6) albinos. To discover the genetic loci that influence age-related retinal degeneration (ARD), a quantitative genetics study was performed with 8-month-old progeny from an intercross between these two strains. METHODS: The thickness of the outer nuclear layer of the retina was used as the quantitative trait. A genome-wide scan was performed with 86 genetic markers at an average distance of 15.7 cM. Map Manager QTX was used to analyze the data. RESULTS: Three highly significant quantitative trait loci (QTLs) were detected on mouse chromosomes (Chrs) 6, 10, and 16. The B6 alleles were protective against ARD in the first two, and the C allele was protective in the third. Several suggestive, weak QTLs were also found, along with a gender-related effect. The strongest and most highly significant QTL on Chr 6 accounted for 30% of the total genetic effect with a LOD score of 13.5. The RPE65/MET450 variant of major influence on constant light-induced retinal degeneration (LRD) in a previous study of these same two mouse strains had no influence on ARD, and only some of the weak, suggestive QTLs influencing ARD were also observed in LRD. CONCLUSIONS: Because none of the ARD QTLs was homologous to human chromosomal loci so far implicated in age-related macular degeneration, each represents a new candidate gene for potential study. The gene represented by the Chr 6 QTL is of particular interest because it has broad influence, very high significance, and a B6 allele that protects against ARD. PMID- 12766040 TI - Molecular characterization of the skate peripherin/rds gene: relationship to its orthologues and paralogues. AB - PURPOSE: A great deal of information about functionally significant domains of a protein may be obtained by comparison of primary sequences of gene homologues over a broad phylogenetic base. This study was designed to identify evolutionarily conserved domains of the photoreceptor disc membrane protein peripherin/rds by analysis of the homologue in a primitive vertebrate, the skate. METHODS: A skate retinal cDNA library was screened using a mouse peripherin/rds clone. The 5' and 3' untranslated regions of the skate peripherin/rds (srds) cDNA were isolated by the rapid amplification of cDNA ends (RACE) approach. The gene structure was characterized by PCR amplification and sequencing of genomic fragments. Northern and Western blot analyses were used to identify srds transcript and protein, respectively. RESULTS: A new homologue of peripherin/rds was identified from the skate retinal cDNA library. SRDS is a glycoprotein with a predicted molecular mass of 40.2 kDa. The srds gene consists of two exons and one small intron and transcribes into a single 6-kb message. Phylogenetic analysis places SRDS at the base of peripherin/rds family and near the division of that group and the branch leading to rds-like and rom-1 genes. SRDS protein is 54.5% identical with peripherin/rds across species. Identity is significantly higher (73%) in the intradiscal domains. Sequence comparison revealed the conservation of all residues that have been shown, on mutation, to associate with retinitis pigmentosa and showed conservation of most residues associated with macular dystrophies. Comparison with ROM-1 and other rds-like proteins revealed the presence of a highly conserved domain in the large intradiscal loop. CONCLUSIONS: Srds represents the skate orthologue of mammalian peripherin/rds genes. Conservation of most of the residues associated with human retinal diseases indicates that these residues serve important functional roles. The high degree of conservation of a short stretch within the large intradiscal loop also suggests an important function for this domain. PMID- 12766042 TI - Distribution of developmental myosin heavy chains in adult rabbit extraocular muscle: identification of a novel embryonic isoform absent in fetal limb. AB - PURPOSE: To identify embryonic and neonatal/fetal myosin heavy chains (MyHCs) in rabbit extraocular muscle (EOM) by electrophoretic and immunochemical analyses and to describe the distribution of these two MyHC isoforms in the endplate zone (EPZ) and the distal and proximal segments of EOM fibers. METHODS: SDS-PAGE and Western blot analysis using monoclonal antibodies (mAbs) against embryonic and neonatal/fetal MyHCs were performed on MyHC isoforms from rabbit adult and neonatal EOM and fetal limb muscles. Immunohistochemical analysis was performed along the entire length of the rabbit superior rectus muscles, using these and other mAbs. RESULTS: Western blot analysis showed that adult rabbit EOM had two embryonic MyHC bands: a weakly stained band that comigrated with the embryonic MyHC from fetal limb muscles, and a strongly stained band of lower electrophoretic mobility for which there was no limb counterpart. Three anti embryonic MyHC mAbs stained muscle fibers, predominantly in the orbital layer, and staining was localized distal and proximal to the EPZ but not in the EPZ itself. There, most fibers expressed the EOM-specific fast MyHC, although some fibers expressed alpha-cardiac MyHC. Anti-neonatal/fetal MyHC mAb failed to stain in Western blot analysis but stained scattered fibers predominantly in the global layer, and there was no specific absence of staining at the EPZ. CONCLUSIONS: There are two electrophoretically distinct isoforms of embryonic MyHCs in adult rabbit EOM. These isoforms are expressed in orbital fibers but are excluded from the EPZ, where EOM-specific fast MyHC is strongly expressed. Neonatal and fetal MyHC is weakly expressed in the EOM, but is not excluded from the EPZ. PMID- 12766043 TI - Information sources and their use by parents of children with ophthalmic disorders. AB - PURPOSE: Parents' input is critical to clinical management in pediatric ophthalmology. The importance of providing parents with appropriate information to enable them to participate effectively is recognized. However, little is known about the range of sources parents use to learn about their child's ophthalmic condition, which sources they find most useful, and how this relates to their understanding. METHODS: Cross-sectional survey of the parents or usual caregivers of children with diverse ophthalmic disorders, diagnosed at least 1 year earlier, who attended pediatric ophthalmology clinics at Great Ormond Street Hospital, London, during 1 week in August 2001. RESULTS: Eighty-nine percent (n = 58) of parents with eligible children participated. Most parents received information from more than one source, with ophthalmologists (79%) and family practitioners (42%) being the two most frequently reported. Family support groups and voluntary organizations (29%) and the Internet (23%) were less commonly cited than anticipated. Parents reported receiving verbal information much more frequently than written information from professionals working with their children. Although 72% (n = 42) of parents could correctly name their child's diagnosis, only 46% (n = 27) were able to describe correctly the nature and impact of the disorder(s). Ophthalmologists were ranked as the most important source overall. CONCLUSIONS: The findings emphasize the key role of ophthalmic professionals in improving parental education directly, as well their responsibilities and opportunities to do so through supporting and shaping information provision through other sources, especially colleagues in primary care and the Internet. PMID- 12766044 TI - Lutein and zeaxanthin status and risk of age-related macular degeneration. AB - PURPOSE: To investigate the relation between plasma concentrations of lutein and zeaxanthin and age-related macular degeneration in a group of elderly men and women. METHODS: The Wisconsin Age-Related Maculopathy Grading System was used to grade features of early and late macular degeneration in 380 men and women, aged 66 to 75 years, from Sheffield, United Kingdom. Fasting blood samples were taken to assess plasma concentrations of lutein and zeaxanthin. RESULTS: Risk of age related macular degeneration (early or late) was significantly higher in people with lower plasma concentrations of zeaxanthin. Compared with those whose plasma concentrations of zeaxanthin were in the highest third of the distribution, people whose plasma concentration was in the lowest third had an odds ratio for risk of age-related macular degeneration of 2.0 (95% confidence interval [CI] 1.0 4.1), after adjustment for age and other risk factors. Risk of age-related macular degeneration was increased in people with the lowest plasma concentrations of lutein plus zeaxanthin (odds ratio [OR] 1.9, 95% CI 0.9-3.5) and in those with the lowest concentrations of lutein (OR 1.7, 95% CI 0.9-3.3), but neither of these relations was statistically significant. CONCLUSIONS: These findings provide support for the view that zeaxanthin may protect against age related macular degeneration. PMID- 12766045 TI - Identification of differentially expressed genes in keratoconus epithelium analyzed on microarrays. AB - PURPOSE: The present study was conducted to investigate differential gene expression in individual samples derived from fresh-frozen human keratoconus and normal corneal epithelium, using gene microarrays. METHODS: Total RNA was extracted (11 keratoconus and 8 normal samples), labeled, and hybridized to microarrays (GeneChip; Affymetrix, Inc., Santa Clara, CA). GeneChip data were validated by verifying the expression profiles of 10 genes by real-time PCR and by recalculation using dChip software (Wong Laboratory, Department of Biostatistics, Harvard School of Public Health, Boston, MA). Furthermore, 3 of the 10 encoded proteins were stained by immunohistochemistry. RESULTS: In comparison with normal cornea, the expression of 471 of the 5600 genes on the microarrays was changed in the keratoconus samples. This number was reduced to 47 with increased expression and 9 with decreased expression when more stringent selection parameters were applied. These genes are believed to be involved in keratoconus. Two of the candidate genes, lysyl oxidase and tissue inhibitor of metalloproteinase 3, are known to be involved in other eye diseases. Expression profiles were reproduced with the software dChip (Wong Laboratory) and real-time PCR. Increases in keratin 6 and 13 were also detected at the protein level. CONCLUSIONS: Keratoconus epithelium appears to be characterized by massive changes of the cytoskeleton, reduced extracellular matrix remodeling, altered transmembrane signaling, and modified cell-to-cell and cell-to-matrix interactions. Validation of gene expression with dChip analysis and real-time PCR indicates GeneChip to be a valid technique for investigation of epithelium from single dissected corneal samples. Association between alterations at RNA and protein levels was observed for some of the tested candidates. PMID- 12766046 TI - Isolation and characterization of cultured human conjunctival goblet cells. AB - PURPOSE: To isolate and characterize goblet cells from normal human conjunctival tissue to determine whether epidermal growth factor (EGF) receptors are present and whether EGF can influence goblet cell proliferation. METHODS: Goblet cells were isolated from explant cultures established from normal conjunctival tissue harvested from patients during periocular surgery. The cells were grown in RPMI culture medium supplemented with 10% fetal bovine serum and characterized using morphology, histochemistry, indirect immunofluorescence microscopy, molecular biology, and biochemistry. Proliferation was determined with a MTT proliferation assay after exposing goblet cells, which had been serum deprived for 48 hours, to increasing concentrations of epidermal growth factor (EGF; 0-80 ng/mL) for 24 hours. RESULTS: Goblet cells were isolated from conjunctival explants by scraping nongoblet cells from the culture dish. Human goblet cells exhibited positive reactivity with alcian blue-periodic acid Schiff (PAS) reagent, goblet cell specific cytokeratin-7, HPA lectin, and MUC5AC, but negative reactivity to the stratified squamous epithelial cell marker, cytokeratin-4. The mRNA for MUC5AC was detected using RT-PCR. The presence of the EGF receptors EGFR, ErbB2, and ErbB3 was confirmed through Western blot analysis of cell lysates. EGF elicited a concentration-dependent increase in goblet cell proliferation of 160% +/- 0.5%, 188% +/- 0.45%, 293% +/- 1.3%, and 220% +/- 0.5% of control values with 10, 20, 40, and 80 ng/mL EGF, respectively. CONCLUSIONS: Human goblet cells that retain characteristics of goblet cells in vivo can be cultured. EGF receptors are present in human goblet cells, and EGF stimulates their proliferation. PMID- 12766047 TI - MUC16 mucin is expressed by the human ocular surface epithelia and carries the H185 carbohydrate epitope. AB - PURPOSE: H185 antibody has been shown to recognize a carbohydrate epitope on a membrane-associated mucin in the apical surfaces of the corneal and conjunctival epithelia. The distribution of this antibody is altered on the surfaces of conjunctival epithelial cells of dry eye patients. The purpose of this work was to determine whether the H185 antibody recognizes the recently cloned membrane associated mucin MUC16 (formerly CA125 antigen). METHODS: To determine whether ocular surface epithelia express MUC16, the relative expression of the MUC16 mucin gene was determined by real-time PCR on reverse transcription products from RNA isolated from human corneal and conjunctival tissues, as well as from immortalized human corneal-limbal epithelial cell (HCLE) cultures. To determine the distribution of MUC16 mRNA and protein in the ocular surface epithelia, in situ hybridization and immunohistochemistry were performed on sections of corneal and conjunctival epithelia using, respectively, a MUC16 antisense oligoprobe and the antibodies OC125, VK-8, and R16 raised against the MUC16 mucin. Determination of whether MUC1 and MUC16 mucins carry the H185 carbohydrate epitope was achieved with the respective mucins isolated from HCLE protein extracts, using one- or two step immunoprecipitation assays and immunodepletion experiments followed by Western blot analysis. RESULTS: MUC16 mucin transcripts were detected in the human ocular surface epithelia and in corneal cell cultures. MUC16 mRNA and protein localized to the apical cell layers of the cornea and to the suprabasal region of the conjunctival epithelium. In HCLE cultures, MUC16 protein was detected in apical cells of islands of stratified cells. Immunofluorescence microscopy demonstrated exact colocalization of the MUC16 mucin and the H185 carbohydrate epitope in sections of human corneal tissue. Immunoprecipitated MUC16 mucin was recognized by the H185 antibody and vice versa, indicating that MUC16 mucin carries the H185 epitope. Immunodepletion with H185 antibody resulted in no OC125 antibody reactivity. No cross-reactivity between immunoprecipitated MUC1 and the H185 antibody was observed. CONCLUSIONS: This study demonstrates that the membrane-associated mucin MUC16 is expressed by the human ocular surface epithelia and that MUC16 carries the H185 carbohydrate epitope. Future studies on the expression of MUC16 and the characterization of the molecular structure of the H185 carbohydrate epitope will determine their biological significance on the healthy ocular surface and in dry eye syndrome. PMID- 12766048 TI - Mucin gene expression in immortalized human corneal-limbal and conjunctival epithelial cell lines. AB - PURPOSE: The corneal and conjunctival epithelia, which cover the ocular surface, play an important role in preventing pathogen penetrance into the eye and maintaining a wet-surface phenotype by producing highly hydrophilic mucin molecules for their apical surfaces. Ocular surface infections, wounding, and pathologies resulting in dry eye threaten sight and can cause blindness. Understanding the ocular surface defense mechanisms that mucins provide has been hampered by the lack of immortalized human corneal and conjunctival epithelial cell lines that retain mucin gene expression patterns of the native tissue. The purpose of this work was to characterize newly developed immortalized corneal and conjunctival cell lines using mucin gene expression as markers of differentiation. METHODS: The cell lines were derived as described by a previously published process. Primary cultures of corneal-limbal and conjunctival epithelia were sequentially transduced to express a dominant negative p53 protein and a p16(INK4A/Rb)-resistant, mutant cdk4 protein, which enabled the cells to bypass a senescence mechanism recently identified for primary cultures of keratinocytes. These cells were then transduced to express the catalytic subunit of telomerase to permit them to retain their telomeres and divide indefinitely. Cellular morphology and expression of mucin genes in the two cell lines, designated HCLE for the human corneal-limbal line and HCjE for the human conjunctival cell line, were determined after culture on plastic, type I collagen, or Matrigel, in coculture with fibroblasts, and in severe combined immunodeficient (SCID) mice. Expression of the epithelial cell mucins was assayed by reverse transcription, real-time polymerase chain reaction, immunoblot analysis, or immunohistochemistry and compared with expression in native cornea and conjunctiva. RESULTS: When grown in high-calcium medium on plastic and type I collagen, cells of both lines stratified, exhibiting multiple cell layers. In Matrigel, both cell lines formed cell aggregates that contained lumens. In the SCID mice, the conjunctival cell line formed stratified layers under the kidney capsule. The corneal cell line expressed keratins K3 and K12, the keratins that are corneal-epithelial-specific, and both cell lines expressed K19. As in native tissue, the HCLE and HCjE cell lines expressed the membrane-associated mucins, MUC1, -4, and -16, although their levels were generally lower. Levels of MUC4 and -16 mRNA were the most comparable to native tissue, particularly when cultured on plastic. Apical cells of the stratified cultures were the cells that expressed the membrane-associated mucins MUC1 and -16. Goblet-cell-specific MUC5AC mRNA and protein was detected in a small population of HCjE cells only when using type I collagen as a substrate or when cells were cocultured with fibroblasts. Both cell lines produced glycosylated mucins as indicated by binding of H185 antibody, an antibody that recognizes a carbohydrate epitope on mucins. CONCLUSIONS: The immortalized corneal (HCLE) and conjunctival (HCjE) cell lines exhibit the mucin gene expression repertoire of their native epithelia. These cell lines will be useful in determining regulation of ocular surface mucin gene expression and, potentially, goblet cell differentiation. PMID- 12766049 TI - Effect of inhibition of inflammatory mediators on trauma-induced stromal edema. AB - PURPOSE: To determine the specific biochemical pathways involved in the initial phase inflammatory response that causes stromal edema after epithelial debridement of the rabbit cornea. METHODS: Adult New Zealand White rabbit corneas were treated with 2 mM synthetic inhibitor of metalloproteinase (SIMP)-1, 1 mM DFU (a specific cyclooxygenase [COX]-2 inhibitor) in 50/50 dimethyl sulfoxide (DMSO)/Ringer's solution, 300 KIU aprotinin (a serine protease inhibitor), 0.05% or 0.10% IL-1 receptor type II solution, 1 mM gliotoxin (a Ras farnesyltransferase inhibitor), or vehicle alone (the control). These were applied topically in vivo in five doses over a 3-hour period except IL-1 receptor type II, which was applied in vitro. After rabbits were killed, the corneas were mounted in perfusion chambers with the endothelium bathed in a modified Ringer's solution and the epithelium bathed with silicone oil. Corneal thickness was measured with an automatic specular microscope. The corneal thickness typically stabilized 1 hour after mounting. After stabilization, the corneal epithelium was removed with a rotating bristle brush and stromal thickness monitored for 1 hour. Paired control corneas were treated similarly. RESULTS. Stromal swelling after epithelial debridement was significantly less in most treated corneas, compared with untreated controls: 18.4 +/- 5.3 microm vs. 28.6 +/- 7.7 microm (n = 6, P = 0.004); SIMP-1, 18.7 +/- 10.2 microm vs. 34.3 +/- 10.2 microm (n = 7, P = 0.02); DFU, 19.3 +/- 10.2 microm vs. 23.5 +/- 8.4 microm (n = 6, P = 0.01); and IL-1 receptor type II (0.05%), 26.2 +/- 5.6 microm vs. 30.4 +/- 5.6 microm (n = 5, P = 0.03) and (0.10%), 26.6 +/- 5.6 microm vs. 32.1 +/- 7.4 microm (n = 8, P = 0.03). Gliotoxin was not effective (21.5 +/- 8.0 microm vs. 21.9 +/- 6.2 microm; n = 5, P = 0.94). CONCLUSIONS: The reduction of stromal edema after topical administration of the inhibitors demonstrates the involvement of the COX-2 enzyme, the matrix metalloproteinase family, plasminogens, and the IL-1 system in the trauma-induced inflammatory response of the rabbit cornea. The inflammatory process in the cornea associated with trauma can proceed along multiple redundant parallel pathways. PMID- 12766051 TI - The effects of overnight orthokeratology lens wear on corneal thickness. AB - PURPOSE: To investigate corneal thickness changes during overnight orthokeratology with reverse-geometry rigid gas-permeable (RGP) contact lenses worn over a 3-month period. METHODS: Eighteen young adult subjects with low myopia (10 on a 0-20 scale). A small but significant learning effect was seen over the two baseline measures for CIGTS VF score and MD. CIGTS VF score and MD correlate highly (r = -0.93); both have high test-retest correlation (0.83 and 0.91, respectively). Variables associated with greater baseline VF loss for both CIGTS VF score and MD include (probabilities for VF only): male sex (P = 0.018), black race (P 6 weeks duration experienced a significant clinical benefit when treated with Dermagraft versus patients treated with conventional therapy alone. With regard to complete wound closure by week 12, 30.0% (39 of 130) of Dermagraft patients healed compared with 18.3% (21 of 115) of control patients (P = 0.023). The overall incidence of adverse events was similar for both the Dermagraft and control groups, but the Dermagraft group experienced significantly fewer ulcer related adverse events. CONCLUSIONS: The data from this study show that Dermagraft is a safe and effective treatment for chronic diabetic foot ulcers. PMID- 12766098 TI - Changes in aerobic capacity and visceral fat but not myocyte lipid levels predict increased insulin action after exercise in overweight and obese men. AB - OBJECTIVE: To examine the effect of moderate intensity physical activity on the interactions between central abdominal adiposity, myocyte lipid content, and insulin action in overweight and obese, sedentary men. RESEARCH DESIGN AND METHODS: Myocyte lipid (biochemical triglyceride and long-chain acyl CoA [LCAC] from vastus lateralis biopsy and soleus and tibialis anterior intramyocellular lipid by (1)H-magnetic resonance spectroscopy), regional body and abdominal fat (dual-energy X-ray absorptiometry and magnetic resonance imaging), serum lipids, insulin action (hyperinsulinemic-euglycemic clamp), and substrate oxidation were measured in 18 nondiabetic, sedentary, and overweight to obese men (aged 37.4 +/- 1.3 years and BMI 30.9 +/- 0.7 kg/m(2), range 26.4-37.6) at baseline, after the first two to four bouts of aerobic exercise (55-70% of VO(2max) for 40 min/session), and at completion of 4.1 +/- 0.2 exercise sessions/week for 9.7 +/- 0.5 weeks (postexercise measurements performed 24-36 h after the last exercise bout). RESULTS: Mean whole body insulin-stimulated glucose uptake and basal fat oxidation rate increased 16 and 41%, respectively, after two to four bouts of exercise, without further increase at program end. Mean aerobic capacity increased 11%, and central abdominal fat decreased 5% at program end, but myocyte lipid levels were not significantly changed. Posttraining increases in insulin stimulated glucose uptake were predicted by increase in aerobic capacity (r = 0.726, P = 0.001) and magnitude of reduction in visceral fat (r = -0.544, P = 0.02) and not by changes in myocyte lipid or LCAC levels. CONCLUSIONS: These results suggest that in overweight and obese sedentary men, increase in insulin sensitivity with moderate intensity exercise is predicted by improvement in aerobic capacity and reduction in visceral fat but is independent of myocyte triglyceride or LCAC levels. PMID- 12766099 TI - Antihyperglycemic effect of oolong tea in type 2 diabetes. AB - OBJECTIVE: To determine the efficacy of oolong tea for lowering plasma glucose in type 2 diabetic patients in Miaoli, Taiwan. RESEARCH DESIGN AND METHODS: A total of 20 free-living subjects who had type 2 diabetes and took hyperglycemic drugs as prescribed were enrolled in the present study. Subjects consumed oolong tea (1,500 ml) or water for 30 days each in a randomized crossover design. Tea was not consumed for 14 days prior to treatments. RESULTS: Relative to initial concentrations, oolong tea markedly lowered concentrations of plasma glucose (from 229 +/- 53.9 to 162.2 +/- 29.7 mg/dl, P < 0.001) and fructosamine (from 409.9 +/- 96.1 to 323.3 +/- 56.4 micromol/l, P < 0.01), whereas the water control group had not changed (208.7 +/- 61.0 vs. 232.3 +/- 63.1 mg/dl for glucose and from 368.4 +/- 85.0 to 340.0 +/- 76.1 micromol/l for fructosamine). CONCLUSIONS: Oolong tea may be an effective adjunct to oral hypoglycemic agents in the treatment of type 2 diabetes. PMID- 12766100 TI - A simple meal plan emphasizing healthy food choices is as effective as an exchange-based meal plan for urban African Americans with type 2 diabetes. AB - OBJECTIVE: To compare a simple meal plan emphasizing healthy food choices with a traditional exchange-based meal plan in reducing HbA(1c) levels in urban African Americans with type 2 diabetes. RESEARCH DESIGN AND METHODS: A total of 648 patients with type 2 diabetes were randomized to receive instruction in either a healthy food choices meal plan (HFC) or an exchange-based meal plan (EXCH) to compare the impact on glycemic control, weight loss, serum lipids, and blood pressure at 6 months of follow-up. Dietary practices were assessed with food frequency questionnaires. RESULTS: At presentation, the HFC and EXCH groups were comparable in age (52 years), sex (65% women), weight (94 kg), BMI (33.5), duration of diabetes (4.8 years), fasting plasma glucose (10.5 mmol/l), and HbA(1c) (9.4%). Improvements in glycemic control over 6 months were significant (P < 0.0001) but similar in both groups: HbA(1c) decreased from 9.7 to 7.8% with the HFC and from 9.6 to 7.7% with the EXCH. Improvements in HDL cholesterol and triglycerides were comparable in both groups, whereas other lipids and blood pressure were not altered. The HFC and EXCH groups exhibited similar improvement in dietary practices with respect to intake of fats and sugar sweetened foods. Among obese patients, average weight change, the percentage of patients losing weight, and the distribution of weight lost were comparable with the two approaches. CONCLUSIONS: Medical nutrition therapy is effective in urban African Americans with type 2 diabetes. Either a meal plan emphasizing guidelines for healthy food choices or a low literacy exchange method is equally effective as a meal planning approach. Because the HFC meal plan may be easier to teach and easier for patients to understand, it may be preferable for low-literacy patient populations. PMID- 12766101 TI - Two-hour insulin determination improves the ability of abdominal fat measurement to identify risk for the metabolic syndrome. AB - OBJECTIVE: Visceral obesity is shown to be a predictor of morbidity and mortality. We evaluated the association of measurements of generalized adiposity and visceral fat area (VFA), with abnormalities of metabolic syndrome (MS). RESEARCH DESIGN AND METHODS: Seventy-six women (47.9 +/- 9.2 years) with BMI of 38.7 +/- 5.4 kg/m(2) underwent anthropometric measurements, laboratory procedures, bioeletrical impedance, and abdominal computed tomography (CT) scan. Diagnosis of MS was based on the presence of abdominal obesity and at least two of the following components: hypertension, dyslipidemia, and glucose intolerance and/or hyperinsulinemia. RESULTS: BMI was correlated with both components of adipose tissue--subcutaneous (r = 0.66, P < 0.01) and VFA (r = 0.33, P < 0.02)- and leptin levels (r = 0.38, P < 0.01). In contrast, VFA was correlated with 2-h glucose and insulin levels (r = 0.32 and 0.35, P < 0.05, respectively), triglyceride, HDL cholesterol, and uric acid (r = 0.33, -0.34 and 0.24, P < 0.05, respectively). Subjects with high VFA, matched for BMI, showed greater plasma glucose area under the curve (621 +/- 127 vs. 558 +/- 129 mg x h(-1) x dl(-1), P < 0.05), 2-h insulin (804 +/- 599 vs. 579 +/- 347 pmol/l, P < 0.05), and uric acid levels (0.33 +/- 0.07 vs. 0.26 +/- 0.06 mmol/l, P < 0.05) than subjects with low VFA. In logistic regression analysis, waist circumference, VFA, and 2-h insulin were identified as independent predictors of MS. Receiver operating characteristic curve analysis pointed out the values of 104 cm for waist circumference (58.1% specificity, 84.1% sensitivity), 158.5 cm(2) for VFA (78.1% specificity, 52.3% sensitivity), and 559.8 pmol/l for 2-h insulin (71.9% specificity, 69.8% sensitivity); the presence of at least two of the three variables resulted in a degree of concordance of 76%. CONCLUSIONS: While BMI was unable to differentiate between obese people and those at higher risk for MS, abdominal fat was shown to be associated with its metabolic abnormalities. The usefulness of abdominal fat in the identification of high-risk subjects may be improved when combined with 2-h insulin determination. PMID- 12766102 TI - The prevalence of and factors associated with diabetic retinopathy in the Australian population. AB - OBJECTIVE: To determine the prevalence and factors associated with diabetic retinopathy in the Australian population and to estimate the time difference between disease onset and clinical diagnosis of type 2 diabetes. RESEARCH DESIGN AND METHODS: The Australian Diabetes, Obesity and Lifestyle study (AusDiab) included 11,247 adults aged > or =25 years in 42 randomly selected areas of Australia. Retinopathy was assessed in participants identified as having diabetes (based on self-report and oral glucose tolerance test), impaired fasting glucose, and impaired glucose tolerance and in a random sample with normal glucose tolerance. Data were available for 2,177 participants. RESULTS: Overall, 15.3% of those with diabetes had retinopathy. The prevalence of retinopathy was 21.9% in those with known type 2 diabetes (KDM) and 6.2% in those newly diagnosed (NDM). The prevalence of proliferative diabetic retinopathy (PDR) was 2.1% in those with KDM. No cases of PDR were found in those with NDM. Untreated vision threatening retinopathy (presence of PDR or macular edema) was present in 1.2% (n = 4). Factors associated with retinopathy were duration of diabetes, HbA(1c), and systolic blood pressure. Using linear extrapolation of the prevalence of retinopathy with diabetes duration, the onset of diabetes in this population was approximately the time of diagnosis. CONCLUSIONS: This is one of the first national studies of diabetic retinopathy in a developed country. The prevalence of retinopathy was similar to that in other population-based studies. Vision threatening retinopathy was relatively rare; however, four untreated cases were identified. Regular screening for diabetic retinopathy and more aggressive management of modifiable risk factors could reduce the numbers of people who develop vision-threatening retinopathy. PMID- 12766103 TI - A randomized clinical trial comparing breakfast, dinner, or bedtime administration of insulin glargine in patients with type 1 diabetes. AB - OBJECTIVE: Insulin glargine (Lantus), a long-acting human insulin analog, provides effective glycemic control when administered at bedtime. This open label, randomized, parallel group, multicenter study investigated whether insulin glargine is equally effective if administered before breakfast, before dinner, or at bedtime. RESEARCH DESIGN AND METHODS: Patients with type 1 diabetes on basal bolus therapy (n = 378, 18-68 years, HbA(1c) 5.5-9.8%) were treated with once daily individually titrated insulin glargine in combination with prandial insulin lispro for 24 weeks. RESULTS: Baseline characteristics were similar in the three groups (overall age 40.9 +/- 11.9 years, diabetes duration 17.3 +/- 11.5 years). Median total daily insulin dose was similar at baseline (0.65, 0.65, and 0.66 IU/kg for breakfast, dinner, and bedtime, respectively) and remained relatively constant over the study period; however, the insulin glargine-to-total insulin dose ratio increased more in the breakfast group than in the dinner and bedtime groups. A similar reduction of adjusted mean HbA(1c) from baseline to end point occurred in all patients (7.6-7.4, 7.6-7.5, and 7.6-7.5% for breakfast, dinner, and bedtime, respectively), and a similar percentage achieved HbA(1c) <7.0% at end point in all groups (29.5, 29.8, and 25.8%, respectively). The 24-h blood glucose profiles in relation to injection time were similar in all groups. The incidences of total symptomatic and severe hypoglycemia did not differ between the three treatment groups; however, nocturnal hypoglycemia occurred in significantly fewer patients in the breakfast group (59.5%) compared with the dinner (71.9%) and bedtime (77.5%) groups (P = 0.005). CONCLUSIONS: These data suggest that insulin glargine, in combination with insulin lispro, is safe and effective when administered before breakfast, before dinner, or at bedtime. PMID- 12766104 TI - Inflammatory markers, adiponectin, and risk of type 2 diabetes in the Pima Indian. AB - OBJECTIVE: To examine the association between adiponectin, a known predictor of diabetes in Pima Indians, and markers of inflammation and endothelial function in nondiabetic subjects and to assess whether these markers predict later diabetes in a case-control study within a longitudinal health study in Pima Indians. RESEARCH DESIGN AND METHODS: Participants with normal glucose tolerance at baseline were selected. Case subjects (who later developed type 2 diabetes), and control subjects (n = 71 pairs) were matched for BMI, age, and sex. Adiponectin, C-reactive protein (CRP), interleukin (IL)-6, tumor necrosis factor-alpha, phospholipase A2 (sPLA2), soluble E-selectin (SE-selectin), soluble intracellular adhesion molecule-1, soluble vascular adhesion molecule-1, and von Willebrand factor (vWF) were measured in baseline samples. RESULTS: Adiponectin was negatively correlated with CRP (r = -0.25, P < 0.05), IL-6 (r = -0.20, P < 0.05), sPLA2 (r = -0.22, P < 0.05), and SE-selectin (r = -0.20, P < 0.05). CRP and IL-6 did not predict diabetes. Only vWF predicted the development of diabetes (incidence rate ratio 0.67 for a 1-SD difference, 95% CI 0.41-1.00, P = 0.05), but this was not significant after adjustment for age, glucose, HbA(1c), waist circumference, and fasting insulin (hazard rate ratio 0.73, 95% CI 0.46-1.16, P = 0.18). CONCLUSIONS: Adiponectin is negatively correlated with markers of inflammation in vivo. In case and control subjects matched for BMI, with the exception of vWF, none of the inflammatory markers predicted diabetes. Adiponectin may be the link between adiposity, inflammation, and type 2 diabetes. PMID- 12766105 TI - Type 2 diabetes and subsequent incidence of breast cancer in the Nurses' Health Study. AB - OBJECTIVE: Hyperinsulinemia may promote mammary carcinogenesis. Insulin resistance has been linked to an increased risk of breast cancer and is also characteristic of type 2 diabetes. We prospectively evaluated the association between type 2 diabetes and invasive breast cancer incidence in the Nurses' Health Study. RESEARCH DESIGN AND METHODS: A total of 116,488 female nurses who were 30-55 years old and free of cancer in 1976 were followed through 1996 for the occurrence of type 2 diabetes and through 1998 for incident invasive breast cancer, verified by medical records and pathology reports. RESULTS: During 2.3 million person-years of follow-up, we identified 6,220 women with type 2 diabetes and 5,189 incident cases of invasive breast cancer. Women with type 2 diabetes had a modestly elevated incidence of breast cancer (hazard ratio [HR] = 1.17; 95% CI 1.01-1.35) compared with women without diabetes, independent of age, obesity, family history of breast cancer, history of benign breast disease, reproductive factors, physical activity, and alcohol consumption. This association was apparent among postmenopausal women (1.16; 0.98-1.62) but not premenopausal women (0.83; 0.48-1.42). The association was predominant among women with estrogen receptor-positive breast cancer (1.22; 1.01-1.47). CONCLUSIONS: Women with type 2 diabetes may have a slightly increased risk of breast cancer. PMID- 12766106 TI - Intensified blood glucose monitoring improves glycemic control in stable, insulin treated veterans with type 2 diabetes: the Diabetes Outcomes in Veterans Study (DOVES). AB - OBJECTIVE: To examine the effect of intensified self-monitored blood glucose (SMBG) testing on glycemic control. RESEARCH DESIGN AND METHODS: Subjects with stable, insulin-treated type 2 diabetes performed SMBG using an electronic blood glucose meter before all meals and at bedtime for 8 weeks. Baseline data were collected on demographics, clinical characteristics, diet, and exercise. HbA(1c) was measured at baseline, at 4 weeks, and at 8 weeks. After the intensified monitoring period, subjects resumed their usual monitoring. HbA(1c) was then measured at 24, 37, and 52 weeks. Multivariate linear regression was used to determine the effect of monitoring on glycemic control. RESULTS: A total of 201 subjects completed the monitoring period. The baseline HbA(1c) (8.10 +/- 1.67%) decreased during the monitoring period by 0.30 +/- 0.68% (P < 0.001) at 4 weeks and by 0.36 +/- 0.88% (P < 0.001) at 8 weeks. Although entry HbA(1c) and compliance independently predicted the week 8 HbA(1c) (r = 0.862, P < 0.001), standardized regression analysis found that compliance with the SMBG protocol influenced the week 8 HbA(1c) more than age, sex, BMI, exercise level, carbohydrate consumption, or treatment intensity at baseline. However, SMBG benefited only subjects whose testing compliance exceeded 75% or with an entry HbA(1c) >8.0%. Decreases in HbA(1c) (-0.31 +/- 1.17%, P = 0.001) persisted in the 159 subjects followed for 52 weeks. CONCLUSIONS: Intensified blood glucose monitoring improved glycemic control in a large cohort of stable, insulin-treated veterans with type 2 diabetes. SMBG provided a strong stimulus for improved self care resulting in clinically important and sustained reductions in HbA(1c). PMID- 12766107 TI - Prevalence and major risk factors of reduced flow volume in lower extremities with normal ankle-brachial index in Japanese patients with type 2 diabetes. AB - OBJECTIVE: To clarify the prevalence and major risk factors of reduced flow volume in lower extremities with normal ankle-brachial index (ABI) in Japanese patients with type 2 diabetes. RESEARCH DESIGN AND METHODS: We recruited 208 consecutive type 2 diabetic patients and 33 age-matched nondiabetic subjects (control group) admitted to our hospital. Thirty-two of the patients had low ABI (<0.90) and intermittent claudication (peripheral arterial disease [PAD] group), and 176 patients had normal ABI (>0.9) (non-PAD group). We evaluated flow volume and resistive index, as an index of arterial resistance to blood flow, at the popliteal artery using gated two-dimensional cine-mode phase-contrast magnetic resonance imaging. RESULTS: Simple linear regression analysis showed a negative correlation between resistive index and total flow volume in the non-PAD group (r = -0.714, P < 0.001). We defined the means +/- 2 SD of these parameters in the control group as the normal range; abnormal resistive index was >1.017, and abnormal flow volume was <50.8 ml/min. The non-PAD group was divided according to the levels of these parameters: 80 patients had both normal resistive index and normal flow volume (normal group); of 96 patients with higher resistive index, 63 had normal flow volume (borderline group) and 33 had reduced flow volume (reduced group). Multiple regression analysis demonstrated that the major risk factors for reduced flow volume were age, hypertension, and diabetic nephropathy (r(2) = 0.303, P < 0.001). CONCLUSIONS: The prevalence of patients without PAD with reduced flow volume in the lower extremities was 16% (n = 33) and comparable with that of patients with PAD with intermittent claudication (n = 32), suggesting that increase in arterial resistance to blood flow may be one of the major causes of lower extremity arterial disease in Japan. PMID- 12766108 TI - Age- and sex-specific prevalence of diabetes and impaired glucose regulation in 11 Asian cohorts. AB - OBJECTIVE: To report the age- and sex-specific prevalence of diabetes and impaired glucose regulation (IGR) according to revised World Health Organization criteria for diabetes in Asian populations. RESEARCH DESIGN AND METHODS: We performed 11 studies of 4 countries, comprising 24,335 subjects (10,851 men and 13,484 women) aged 30-89 years who attended the 2-h oral glucose tolerance test and met the inclusion criteria for data analysis. RESULTS: The prevalence of diabetes increased with age and reached the peak at 70-89 years of age in Chinese and Japanese subjects but peaked at 60-69 years of age followed by a decline at the 70 years of age in Indian subjects. At 30-79 years of age, the 10-year age specific prevalence of diabetes was higher in Indian than in Chinese and Japanese subjects. Indian subjects also had a higher prevalence of IGR in the younger age groups (30-49 years) compared with that for Chinese and Japanese subjects. Impaired glucose tolerance was more prevalent than impaired fasting glycemia in all Asian populations studied for all age-groups. CONCLUSIONS: Indians had the highest prevalence of diabetes among Asian countries. The age at which the peak prevalence of diabetes was reached was approximately 10 years younger in Indian compared with Chinese and Japanese subjects. Diabetes and IGR will be underestimated in Asians based on the fasting glucose testing alone. PMID- 12766109 TI - Prevalence of the metabolic syndrome among Omani adults. AB - OBJECTIVE: To estimate the prevalence of the metabolic syndrome by age and sex in the Omani population as defined by the third report of the National Cholesterol Education Program Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel III [ATP III]) of North America. RESEARCH DESIGN AND METHODS: We analyzed data from a cross-sectional survey conducted in 2001 containing a probability random sample of 1,419 Omani adults aged > or =20 years living in the city of Nizwa. The metabolic syndrome, defined by the ATP III, was defined as having three or more of the following abnormalities: waist circumference >102 cm in men and >88 cm in women, serum triglycerides > or =150 mg/dl (1.69 mmol/l), HDL cholesterol <40 mg/dl (1.04 mmol/l) in men and <50 mg/dl (1.29 mmol/l) in women, systolic blood pressure > or =130 mmHg and/or diastolic > or =85 mmHg or on treatment for hypertension, and fasting serum glucose > or =110 mg/dl (6.1 mmol/l) or on treatment for diabetes. RESULTS: The age-adjusted prevalence of the metabolic syndrome was 21.0%. The crude prevalence was slightly lower (17.0%). The age-adjusted prevalence was 19.5% among men and 23.0% among women (P = 0.236). Low HDL cholesterol was the most common component (75.4%) of the metabolic syndrome among the study population followed by abdominal obesity (24.6%). Abdominal obesity was markedly higher in women (44.3%) than in men (4.7%). CONCLUSIONS: The prevalence of the metabolic syndrome in Oman is similar to that in developed countries. Future prevention and control strategies should not overlook the importance of noncommunicable disease risk factors in rapidly developing countries. PMID- 12766110 TI - High incidence of childhood type 1 diabetes in Liguria, Italy, from 1989 to 1998. AB - OBJECTIVE: Assessing updated incidence of type 1 diabetes in 0- to 14-year-old children in Liguria, a Northwest region of Italy. RESEARCH DESIGN AND METHODS: Incident cases were recorded prospectively from 1989 to 1998. Incidence rates (IRs) were standardized to the 1999 world population using the direct method. The independent effect of sex, age, residence, and calendar year was estimated with Poisson regression model. The degree of ascertainment was calculated in accordance to capture/recapture method. RESULTS: During 10 full calendar years, 219 new cases of type 1 diabetes in children were diagnosed in Liguria. The standardized IR over the 10-year period was 12.56 cases per 100,000 per year (95% CI 11.0-14.3). The sex-specific IR among men and women was 14.15 and 10.88, respectively. The age-specific IR was higher in the 10- to 14-year-old age-group (15.01/100,000) than in 0- to 4-year-old age-group (9.01/100,000) and in the 5- to 9-year-old age-group (13.03/100,000). CONCLUSIONS: The IR of type 1 diabetes in Liguria is among the highest in Southern Europe and approaches IRs of Northern European countries. In particular it is much higher than those reported in the surrounding Italian regions except for Sardinia. Therefore, the geographical distribution of type 1 diabetes does not seem to reflect the simple North-South gradient reported in several previous works. PMID- 12766111 TI - The health care costs of diabetic peripheral neuropathy in the US. AB - OBJECTIVE: Peripheral neuropathy is common among people with diabetes and can result in foot ulceration and amputation. The aim of this study was to quantify the annual medical costs of peripheral neuropathy and its complications among people with type 1 and type 2 diabetes in the U.S. RESEARCH DESIGN AND METHODS: A cost-of-illness model was used to estimate the numbers of diabetic individuals in the U.S. who have diabetic peripheral neuropathy (DPN) and/or neuropathic foot ulcers (both those with no deep infection and those accompanied by cellulitis or osteomyelitis) at a given point in time, and/or a toe, foot, or leg amputation during a year. Prevalence and incidence rates were estimated from published studies and applied to the general U.S. population. All costs were estimated in 2001 U.S. dollars. In a sensitivity analysis, we varied the rates of complications to assess the robustness of the cost estimates. RESULTS: The annual costs of DPN and its complications in the U.S. were 0.8 billion US dollars (type 1 diabetes), 10.1 billion US dollars (type 2 diabetes), and 10.9 billion US dollars (total). After allowing for uncertainty in the point estimates of complication rates, the range of costs were between 0.3 and 1.0 billion US dollars (type 1 diabetes), 4.3b and 12.7 billion US dollars (type 2 diabetes), and 4.6 and 13.7 billion US dollars (type 1 and type 2 diabetes). CONCLUSIONS: The total annual cost of DPN and its complications in the U.S. was estimated to be between 4.6 and 13.7 billion US dollars. Up to 27% of the direct medical cost of diabetes may be attributed to DPN. PMID- 12766113 TI - Treating hypertension in diabetic nephropathy. AB - OBJECTIVE: Control of hypertension in patients with diabetic nephropathy improves mortality and slows progression to end-stage renal disease. However, blood pressure is difficult to treat; multiple drug combination therapy is required and treatment algorithms to establish this are lacking. We used a stepped-care algorithm, centered on maximum doses of an ACE inhibitor or angiotensin II receptor blocker, to treat hypertension according to American Diabetes Association recommended blood pressure target goals (<130/80 mmHg) in patients with diabetic nephropathy. RESEARCH DESIGN AND METHODS: We treated 49 consecutive patients with diabetes (13 with type 1 and 36 with type 2), diabetic nephropathy, and proteinuria > or =500 mg/24 h with a stepped-care blood pressure treatment algorithm. The level of blood pressure control achieved at most recent follow-up was assessed. RESULTS: Patients were followed for a median of 18 months (range 9 48). Mean blood pressure achieved was 140/75 +/- 23/14 mmHg in patients with type 1 diabetes and 146/76 +/- 22/14 mmHg in patients with type 2 diabetes. Target blood pressure was reached in 16 (33%) patients, 6 of 13 patients with type 1 diabetes and 10 of 36 patients with type 2 diabetes, whereas systolic blood pressure remained above the target level in the remaining patients. There was no difference in baseline blood pressure, proteinuria, or serum creatinine level between patients who were treated to target and those who were not. CONCLUSIONS: Levels of blood pressure control similar to those achieved in clinical trials in diabetic nephropathy were obtained with a stepped-care algorithm. However, in most patients, systolic blood pressure was difficult to control to target despite the use of multiple drug combination therapy. PMID- 12766112 TI - Cost effectiveness of statin therapy for the primary prevention of major coronary events in individuals with type 2 diabetes. AB - OBJECTIVE: To assess the cost and cost effectiveness of hydroxymethylglutaryl (HMG)-CoA reductase inhibitor (statin) therapy for the primary prevention of major coronary events in the U.S. population with diabetes and LDL cholesterol levels > or =100 mg/dl, especially in the population with LDL cholesterol levels 100-129 mg/dl. RESEARCH DESIGN AND METHODS: Analyses were performed using population estimates from National Health and Nutrition Examination Survey (NHANES)-III, cost estimates from a health system perspective, statin LDL lowering effectiveness from pivotal clinical trials, and treatment effectiveness from the diabetic subgroup analysis of the Heart Protection Study. RESULTS: There are approximately 8.2 million Americans with diabetes, LDL cholesterol levels > or =100 mg/dl, and no clinical evidence of cardiovascular disease. Each year, statin therapy could prevent approximately 71,000 major coronary events in this population. In the subgroup with LDL cholesterol levels 100-129 mg/dl, the annual cost of statin treatment ranges from 600 to 1,000 US dollars per subject. In the population with LDL cholesterol levels > or =130 mg/dl, the annual cost ranges from 700 to 2,100 US dollars. Annual incremental cost per subject, defined as the cost of statin treatment plus the cost of major coronary events with statin treatment minus the cost of major coronary events without statin treatment, ranges from 480 to 950 US dollars in the subgroup with LDL cholesterol levels 100-129 mg/dl and from 590 to 1,920 US dollars in the population with LDL cholesterol levels > or =130 mg/dl. CONCLUSIONS: Statin therapy for the primary prevention of major coronary events in subjects with type 2 diabetes and LDL cholesterol levels 100-129 mg/dl is affordable and cost effective relative to statin therapy in subjects with higher LDL cholesterol levels. PMID- 12766114 TI - Identification and management of diabetic nephropathy in the diabetes clinic. AB - OBJECTIVE: To examine the prevalence and management of diabetic nephropathy in a diabetes clinic. RESEARCH DESIGN AND METHODS: Characteristics of nephropaths identified by existing screening practice (phase I, albuminuria >20 mg/l in three separate urine samples), were compared with those identified by a nurse-led management program (phase II, in which screening for nephropathy was based on albumin-to-creatinine ratio in a single random urine specimen). RESULTS: In phase I, 644 patients attended a diabetes clinic over a 6-month period. Microalbuminuria results were available for 485 patients (75%). A total of 115 patients were identified as nephropaths (prevalence 17.8%). Of these patients, 91% had type 2 diabetes. During phase II, prospective analysis of urinary albumin to-creatinine ratio was carried out in 880 patients over 8 months. A total of 174 patients were identified as nephropaths (prevalence 20%). Of these, 134 patients had been identified by existing screening protocols. Forty had no previous record of microalbuminuria and were therefore newly identified by prospective screening. Systolic blood pressure guidelines were met in only 31% of all known nephropaths and 26.5% of newly diagnosed nephropaths. Diastolic blood pressure guidelines were met in 36% of all known and 38% of newly diagnosed nephropaths. In the patient group of known nephropaths from phases I and II, 62% were prescribed ACE inhibitors (ACEIs) or angiotensin II receptor (AIIR) antagonists. In the newly identified nephropathy patient cohort from phase II, 48% used ACEIs or AIIR antagonists. CONCLUSIONS: Introduction of a nurse-led management program significantly improved detection of nephropathy. We are currently evaluating its impact on clinical management. PMID- 12766115 TI - Steroids in adult men with type 1 diabetes: a tendency to hypogonadism. AB - OBJECTIVE: To compare steroids and their associations in men with type 1 diabetes and healthy control subjects. RESEARCH DESIGN AND METHODS: We studied 52 adult men with type 1 diabetes without microvascular complications, compared with 53 control subjects matched for age and BMI. Steroids and their binding globulins were assessed in a single venous blood sample and a 24-h urine sample. RESULTS: In adult men with type 1 diabetes, total testosterone did not differ from healthy control subjects, but sex hormone-binding globulin (SHBG) (42 [14-83] vs. 26 [9 117] nmol/l, P < 0.001), cortisol-binding globulin (CBG; 0.87 +/- 0.17 vs. 0.73 +/- 0.10 nmol/l, P < 0.001), and cortisol levels (0.46 +/- 0.16 vs. 0.39 +/- 0.14 nmol/l, P < 0.01) were higher. The free testosterone index was lower (60 [17-139] vs. 82 [24-200], P < 0.001), and the calculated free testosterone was slightly lower (497 [115] vs. 542 [130], P < 0.064), but the pituitary-gonadal axis was not obviously affected in type 1 diabetes. The calculated free serum cortisol was not different, and 24-h urinary free cortisol excretion was lower in type 1 diabetes (121 [42-365] vs. 161 [55-284] nmol/24 h, P < 0.009). Testosterone was mainly associated with SHBG. Estimated portal insulin was a contributor to SHBG in control subjects but not in type 1 diabetes. Cortisol was associated with CBG. HbA(1c) contributed to CBG in men with diabetes but not in control subjects, whereas estimated portal insulin did not contribute. CONCLUSIONS: Adult men with fairly controlled type 1 diabetes without complications who are treated with subcutaneous insulin have a tendency to hypogonadism, as reflected by lower free testosterone levels in the presence of similar total testosterone levels and higher SHBG levels. PMID- 12766116 TI - Kobberling type of familial partial lipodystrophy: an underrecognized syndrome. AB - OBJECTIVE: The phenotypic expression of partial lipodystrophy is present in two familial syndromes: familial partial lipodystrophy type 1 (FPLD1), with fat loss from the extremities, and central obesity and FPLD type 2, with fat loss from the extremities, abdomen, and thorax. The latter disorder is associated with mutations in the LMNA gene. FPLD1 is thought to be rare. Here, we report 13 subjects with FPLD1, suggesting that this syndrome is more common than previously thought. RESEARCH DESIGN AND METHODS: Fasting glucose, plasma lipids, leptin, HbA(1c), and anthropomorphic measurements were evaluated in 13 subjects with clinical features of FPLD1 and are compared with two age-matched control groups, with and without diabetes. RESULTS: Only women with clinical features of FPLD1 have been identified. Although they lack extremity and gluteal subcutaneous fat, they do have truncal obesity. Skinfold thickness on the arm and leg was significantly less than that in control subjects. The ratio of skinfold thickness from abdomen to thigh was significantly higher in subjects, suggesting an easy method for identifying affected patients. FPLD1 subjects also had components of the metabolic syndrome, including hypertension, insulin resistance, and severe hypertriglyceridemia resulting in pancreatitis. Premature coronary artery disease was present in 31% of subjects. None of the subjects had coding mutations in the LMNA gene or in the gene coding for peroxisome proliferator-activated receptor (PPAR)-gamma. CONCLUSIONS: FPLD1 is more common than previously described, but the diagnosis is often missed. Early recognition and intensive treatment of hyperlipidemia and diabetes in FPLD1 is important for prevention of pancreatitis and early cardiovascular disease. PMID- 12766118 TI - Developing a screening program to detect sight-threatening diabetic retinopathy in South India. AB - OBJECTIVE: To develop a screening protocol for detection of sight-threatening diabetic retinopathy in south India. RESEARCH DESIGN AND METHODS: We performed ophthalmic examinations, including posterior segment examination, using indirect ophthalmoscopy to detect sight-threatening retinopathy in patients with diabetes in screening camps targeting a high-risk population. RESULTS: We examined 3,949 persons with diabetes in 32 screening camps over a 13-month period beginning July 2001. Most of the patients (93.6%) were aware of their diabetic status, and 84.2% of those aware of their diabetes status were on treatment. One-fifth of those screened had evidence for any retinopathy; only 6.1% of these persons had evidence of past ophthalmic treatment for retinopathy. Only one-quarter of those diagnosed with worse than mild retinopathy came for follow-up to the base hospital within 2 months. CONCLUSIONS: Screening high-risk groups for sight threatening retinopathy using indirect ophthalmoscopy may be a useful short-term alternative for India until retinal photography becomes affordable. In addition to strategies to improve coverage, strategies for better follow-up of subjects screened also need to be evolved. PMID- 12766117 TI - Correlation between midthigh low-density muscle and insulin resistance in obese nondiabetic patients in Korea. AB - OBJECTIVE: We investigated the link between lipid-rich skeletal muscle, namely low-density muscle, and insulin resistance in Korea. RESEARCH DESIGN AND METHODS: Abdominal adipose tissue areas and midthigh skeletal muscle areas of 75 obese nondiabetic subjects (23 men, 52 women; mean age +/- SD, 41.9 +/- 14.1 years) were measured by computed tomography (CT). The midthigh skeletal muscle areas were subdivided into low-density muscle (0 to +30 Hounsfield units) and normal density muscle (+31 to +100 Hounsfield units). The homeostasis model assessment (HOMA) score was calculated to assess whole-body insulin sensitivity. RESULTS: The abdominal visceral fat area and the midthigh low-density muscle area were found to be well correlated with the HOMA score (r = 0.471, P < 0.01 and r = 0.513, P < 0.01, respectively). The correlation between low-density muscle area and insulin resistance persisted after adjusting for BMI or total body fat mass (r = 0.451, P < 0.01 and r = 0.522, P < 0.01, respectively) and even after adjusting for abdominal visceral fat area (r = 0.399, P < 0.01). CONCLUSIONS: The midthigh low-density muscle area seems to be a reliable determinant of insulin resistance in Korean obese nondiabetic patients. PMID- 12766119 TI - L-arginine-induced vasodilation of the renal vasculature is not altered in hypertensive patients with type 2 diabetes. AB - OBJECTIVE: Diabetes, arterial hypertension, hypercholesterolemia, and aging are associated with endothelial dysfunction in various vasculatures. Endothelium dependent vasodilation of the renal vasculature cannot be easily assessed, but infusion of L-arginine, the substrate of endothelial nitric oxide synthase, leads to an increase in renal plasma flow (RPF) in humans. We have examined the effect of L-arginine infusion on renal hemodynamics in hypertensive patients with type 2 diabetes. RESEARCH DESIGN AND METHODS: Twenty-three elderly patients with type 2 diabetes (age, 65 +/- 6 years; HbA(1c), 7.8 +/- 1.6%) with coexisting arterial hypertension (158 +/- 19/83 +/- 11 mmHg) and elevated cholesterol levels (total cholesterol, 215 +/- 33 mg/dl) were examined. These patients were compared with a young and healthy reference group (n = 20; age, 26 +/- 2 years). The effect of L arginine infusion (100 mg/kg over 30 min) on RPF and glomerular filtration rate were measured using the constant input clearance technique with p-aminohippurate and inulin, respectively. RESULTS: L-arginine infusion similarly influenced renal hemodynamics in patients and reference subjects: RPF increased by 7 +/- 11 and 7 +/- 11% in diabetic and reference subjects, respectively (P = NS). Other parameters of renal hemodynamics such as glomerular filtration rate (5 +/- 5 vs. 4 +/- 4%) and filtration fraction (-1 +/- 8 vs. -1 +/- 9%) were not significantly different between diabetic and reference subjects, too. CONCLUSIONS: L-arginine induced vasodilation of the renal vasculature is not different between a group of hypertensive diabetic patients and a young, healthy reference group. These data were obtained using low-dose L-arginine infusion. PMID- 12766120 TI - On the association between diabetes and mental disorders in a community sample: results from the German National Health Interview and Examination Survey. AB - OBJECTIVE: To determine the relationship between mental disorders and diabetes in a representative community sample. RESEARCH DESIGN AND METHODS: This was a cross sectional study. Data on diabetes and HbA(1c) values were obtained by structured questionnaires and by laboratory assessments. Current psychiatric disorders were diagnosed by a modified version of the Composite International Diagnostic Interview (CIDI). RESULTS: People with diabetes (PWD) were not more likely to meet Diagnostic and Statistical Manual of Psychiatric Disorders, 4th edition (DSM IV) criteria for at least one mental disorder than were individuals without diabetes. However, a different diagnostic pattern occurred compared with the general population: odds ratios (ORs) for anxiety disorders in PWD were higher (OR 1.93, 95% CI 1.19-3.14). Although PWD had higher prevalence rates of affective disorders, the relationship between diabetes and affective disorders was not statistically significant after controlling for age, sex, marital status, and socioeconomic status. In contrast, the relationship between diabetes and anxiety disorders remained significant after controlling for these variables. In contrast to individuals without mental disorders, PWD with affective or anxiety disorders more frequently had adequate glycemic control. CONCLUSIONS: Diabetes was associated with an increased likelihood of anxiety disorders. The association between mental disorders, diabetes, and glycemic control should be evaluated carefully in terms of potentially confounding sociodemographic variables, sample characteristics, and definitions of the disorders. PMID- 12766121 TI - Recent antihyperglycemic prescribing trends for US privately insured patients with type 2 diabetes. AB - OBJECTIVE: The mid-1990s witnessed the introduction of new classes of medications to treat hyperglycemia of type 2 diabetes. There is evidence that these newer classes have found a place in the therapeutic armamentarium, but details of their use patterns are not known. We sought to determine whether antihyperglycemic prescribing patterns changed concurrently with new drug introductions, and whether such changes were related to changes in the underlying patient population. RESEARCH DESIGN AND METHODS: A sample of U.S. privately insured patients with suspected type 2 diabetes was identified from the MarketScan Research Database over the period of 1997-2000. Patients with type 2 diabetes were identified among those continuously enrolled in the database for at least 1 year. Drug therapy episodes were defined by sequential fulfillment of prescriptions implying a continuous supply of a particular drug (or combination) of at least 30 days duration. Univariate analyses were used to explore trends over time in drug prescriptions and patient characteristics. Multivariate logistic regressions were used to isolate the impact of year from other variables on the likelihood of receiving prescriptions for a specific therapy. RESULTS: A total of 232,020 unique diabetic patients had an average of 1.91 diabetes drug therapy episodes between 1997 and 2000. Monotherapy with sulfonylureas decreased, but monotherapy with thiazolidinedione, metformin, and other oral antihyperglycemics increased over time. Combinations of sulfonylureas and metformin; sulfonylureas and thiazolidinedione; metformin and thiazolidinedione; and sulfonylureas, metformin, and thiazolidinedione each increased over the time interval. Insulin monotherapy decreased, as did insulin combination therapy with sulfonylureas. The combination of insulin and metformin increased, whereas insulin and thiazolidinedione was stable. The influence of year on prescribing patterns remained highly significant (P < 0.001) after adjusting for patient characteristics. CONCLUSIONS: Antihyperglycemic prescription patterns in the U.S. have changed in recent years in parallel with, and probably as a direct result of, the introduction of different classes of medications to the marketplace. Overall, the prescribing trend has been away from monotherapy with insulins and sulfonylureas and toward combination therapies, presumably in attempts to reduce hypoglycemic symptoms and to achieve better glucose control. PMID- 12766122 TI - Rapid increase in the use of oral antidiabetic drugs in the United States, 1990 2001. AB - OBJECTIVE: To describe the use of oral antidiabetic drugs for management of type 2 diabetes in the U.S. from 1990 through 2001. RESEARCH DESIGN AND METHODS: Data on oral antidiabetic drugs were derived from two pharmaceutical marketing databases from IMS Health, the National Prescription Audit Plus and the National Disease and Therapeutic Index. RESULTS: In 1990, 23.4 million outpatient prescriptions of oral antidiabetic agents were dispensed. By 2001, this number had increased 3.9-fold, to 91.8 million prescriptions. Glipizide and glyburide, two sulfonylurea medications, accounted for approximately 77% of prescriptions of oral antidiabetic drugs in 1990 and 35.5% of prescriptions in 2001. By 2001, the biguanide metformin (approved in 1995) had captured approximately 33% of prescriptions, and the thiazolidinedione insulin sensitizers (rosiglitazone and pioglitazone marketed beginning in 1999) accounted for approximately 17% of market share. Compared with patients treated in 1990, those in 2001 were proportionately younger and they more often used oral antidiabetic drugs and insulin in combination. Internists and general and family practitioners were the primary prescribers of this class of drugs. CONCLUSIONS: Consistent with the reported increase in the prevalence of type 2 diabetes, the number of dispensed outpatient prescriptions of oral antidiabetic drugs increased rapidly between 1990 and 2001. This period was marked by an increase in the treatment of younger people and the use of oral antidiabetic drugs in combination. With the approval in the last decade of several new types of oral antidiabetic medications with different mechanisms of action, options for management of type 2 diabetes have expanded. PMID- 12766123 TI - Human epidermal growth factor enhances healing of diabetic foot ulcers. AB - OBJECTIVE: To study the healing effect of recombinant human epidermal growth factor (hEGF) on diabetic foot ulcers. RESEARCH DESIGN AND METHODS: A total of 127 consecutive patients were screened and 61 diabetic subjects were recruited into this double-blind randomized controlled study. Predetermined criteria were used for diagnosis and classification of the diabetic wound. The patients were randomized into three groups. All patients attended our Diabetes Ambulatory Care Center every other week for joint consultation with the diabetologist and the podiatrist. Group 1 (control) was treated with Actovegin 5% cream (Actovegin), group 2 with Actovegin plus 0.02% (wt/wt) hEGF, and group 3 with Actovegin plus 0.04% (wt/wt) hEGF. The study end point was the complete closure of the wound. Failure to heal was arbitrarily defined as incomplete healing after 12 weeks. RESULTS: Final data were obtained from 61 patients randomly assigned into three groups. The mean ages of the patients, wound sizes, wound duration, metabolic measurements, and comorbidities were comparable within groups, except that group 3 had more female patients. Mean follow-up for the patients was 24 weeks. Data were cutoff at 12 weeks, and results were analyzed by intention to treat. After 12 weeks, in group 1 (control) eight patients had complete healing, two patients underwent toe amputation, and nine had nonhealing ulcers. In group 2 (0.02% [wt/wt] hEGF) 12 patients experienced wound healing, 2 had toe amputations, and 7 had nonhealing ulcers. Some 20 of 21 patients in group 3 (0.04% [wt/wt] hEGF) showed complete wound healing. Healing rates were 42.10, 57.14, and 95% for the control, 0.02% (wt/wt) hEGF, and 0.04% (wt/wt) hEGF groups, respectively. Kaplan Meier survival analysis suggested that application of cream with 0.04% (wt/wt) hEGF caused more ulcers to heal by 12 weeks and increased the rate of healing compared with the other treatments (log-rank test, P = 0.0003). CONCLUSIONS: Our data support the contention that application of hEGF-containing cream, in addition to good foot care from a multidisciplinary team, significantly enhances diabetic foot ulcer wound healing and reduces the healing time. PMID- 12766125 TI - Association between serum testosterone concentration and carotid atherosclerosis in men with type 2 diabetes. AB - OBJECTIVE: There is evidence to suggest that low concentrations of testosterone are associated with an increased risk of cardiovascular disease in men. The aim of this study was to evaluate the relationship between serum testosterone concentration and carotid atherosclerosis as well as major cardiovascular risk factors in men with type 2 diabetes. RESEARCH DESIGN AND METHODS: Serum free and total testosterone concentrations were measured in 253 consecutive men with type 2 diabetes. The relationships between serum testosterone concentration and carotid atherosclerosis, determined by ultrasonographically evaluated intima media thickness (IMT) and plaque score (PS) in a subgroup of 154 diabetic patients, as well as major cardiovascular risk factors, including age, blood pressure, and lipid concentrations, were evaluated. RESULTS: Inverse correlations were found between free testosterone (F-tes) concentration and IMT (r = -0.206, P = 0.0103) and between F-tes concentration and PS (r = -0.334, P < 0.001). The IMT and PS were significantly greater in patients with lower concentrations of F-tes (<10 pg/ml) than in patients with higher concentrations of F-tes (1.01 +/- 0.29 vs. 0.91 +/- 0.26 mm, P = 0.038; 4.5 +/- 3.8 vs. 2.4 +/- 3.2, P = 0.0003; respectively). An inverse correlation was found between serum F-tes concentration and age (r = -0.420, P < 0.0001). A positive correlation was found between serum F-tes and total cholesterol concentrations (r = 0.145, P = 0.0238). CONCLUSIONS: Serum F-tes concentration is inversely associated with carotid atherosclerosis determined by ultrasonographically evaluated IMT and PS in men with type 2 diabetes. PMID- 12766126 TI - Ulcer recurrence following first ray amputation in diabetic patients: a cohort prospective study. AB - OBJECTIVE: To evaluate the reulceration and reamputation rates in a cohort of diabetic patients following first ray amputation. RESEARCH DESIGN AND METHODS: We evaluated a cohort of 89 diabetic patients, 63 men and 26 women, who underwent first ray amputation in the period from January 2000 to December 2001. The first ray lesions were Wagner grade 2 in 3 patients, Wagner grade 3 in 47 patients, and Wagner grade 4 in 39 patients. Following surgical wound healing, all patients wore special footwear with rocker bottom soles and custom molded insoles and were put on an intensive secondary prevention program. RESULTS: The mean follow-up duration was 16.35 +/- 6.76 months (range 7-28). Fifteen patients developed new ulcerations, with 11 lesions occurring ipsilaterally and 4 contralaterally to the first ray amputation. In seven patients, the new lesion was treated and healed with dressing. Eight patients underwent a new surgical procedure: panmetatarsal head resection in four patients, toe amputation in two patients, a transmetatarsal amputation in one patient, and Lisfranc's amputation in one patient. CONCLUSIONS: In the population studied, the first ray amputation presented a lower reulceration and reamputation rate with respect to that reported in the literature. This finding should therefore be attributed to the follow-up program, which uses shoes with a rocker bottom sole and custom molded insoles and intensive ambulatory check-ups. PMID- 12766127 TI - Percent change in wound area of diabetic foot ulcers over a 4-week period is a robust predictor of complete healing in a 12-week prospective trial. AB - OBJECTIVE: To assess the ability of the 4-week healing rate to predict complete healing over a 12-week period in a large prospective multicenter trial of diabetic patients with foot ulceration. RESEARCH DESIGN AND METHODS: We examined the change in ulcer area over a 4-week period as a predictor of wound healing within 12 weeks in patients who were seen weekly in a prospective, randomized controlled trial. RESULTS: Wound area measurements at baseline and after 4 weeks were performed in 203 patients. The midpoint between the percentage area reduction from baseline at 4 weeks in patients healed versus those not healed at 12 weeks was found to be 53%. Subjects with a reduction in ulcer area greater than the 4-week median had a 12-week healing rate of 58%, whereas those with reduction in ulcer area less than the 4-week median had a healing rate of only 9% (P < 0.01). The absolute change in ulcer area at 4 weeks was significantly greater in healers versus nonhealers (1.5 vs. 0.8 cm(2), P < 0.02). The percent change in wound area at 4 weeks in those who healed was 82% (95% CI 70-94), whereas in those who failed to heal, the percent change in wound area was 25% (15 35; P < 0.001). CONCLUSIONS: The percent change in foot ulcer area after 4 weeks of observation is a robust predictor of healing at 12 weeks. This simple tool may serve as a pivotal clinical decision point in the care of diabetic foot ulcers for early identification of patients who may not respond to standard care and may need additional treatment. PMID- 12766124 TI - Effects of the early ACE inhibition in diabetic nonthrombolyzed patients with anterior acute myocardial infarction. AB - OBJECTIVE: The aim of the present study was to evaluate the clinical efficacy of the ACE inhibitor zofenopril in a cohort of diabetic patients with nonthrombolyzed anterior acute myocardial infarction who were enrolled in the Survival of Myocardial Infarction Long-Term Evaluation (SMILE) trial. RESEARCH DESIGN AND METHODS: Among the overall population of 1,512 patients, 303 (20.0%) had diabetes. The primary end point of this study was the effect of treatment on the 6-week combined occurrence of death and severe congestive heart failure (CHF). Secondary end points included the evaluation of the 6-week rate of major cardiovascular events as well as the 1-year survival rate. RESULTS: After 6 weeks of double-blind treatment, zofenopril significantly reduced both the incidence of the primary end point (8.6 vs. 18.3%; P = 0.019) and the 6-week incidence of severe CHF (0 vs. 7.3%; P = 0.001) in diabetic patients, and the effect was greater than that observed in nondiabetic patients. Conversely, 1-year mortality was significantly reduced among nondiabetic patients (9.1 vs. 13.8%; P = 0.010), whereas in the diabetic population, the decrease did not reach statistical significance (13.7 vs. 16.5%; P = 0.52). CONCLUSIONS: The present data suggest that the clinical outcome of patients with diabetes and myocardial infarction can be significantly improved by early treatment with zofenopril. The lesser effect on 1-year mortality seems to suggest that long-term treatment is probably needed to maintain the benefits of the early ACE inhibition in patients with diabetes. PMID- 12766128 TI - Cutaneous blood flow in type 2 diabetic individuals after an acute bout of maximal exercise. AB - OBJECTIVE: We previously demonstrated a positive association between chronic aerobic exercise and dorsal foot skin blood flow during local heating in type 2 diabetic individuals. Thus, we hypothesized that a prior acute bout of maximal exercise would also have positive effects on postexercise blood flow. RESEARCH DESIGN AND METHODS: Subjects consisted of 32 individuals with type 2 diabetes and 26 nondiabetic control subjects further subdivided based on their physical activity status: diabetic exerciser (DE), diabetic sedentary (DS), control exerciser (CE), or control sedentary. Dorsal foot cutaneous blood flow was measured noninvasively by continuous laser-Doppler assessment at baseline and during local heating to 44 degrees C before and after a maximal bout of cycle exercise. Interstitial nitric oxide (NO) levels were measured concurrently in the foot dorsum. RESULTS: Increases in blood flow and its responsiveness to local heating to 44 degrees C were significantly lower in both diabetic groups compared with CE before maximal exercise, but perfusion responsiveness remained lower in DS subjects only after exercise (P < 0.05). Baseline skin blood flow was not different among groups preexercise, but it was significantly increased postexercise in DE subjects only. Interstitial NO levels were not significantly different at either time. At baseline, groups differed only in HbA(1c), fasting serum glucose, HDL cholesterol, and insulin resistance (homeostasis model assessment method). CONCLUSIONS: All diabetic individuals exhibit a blunted responsiveness of cutaneous blood flow with local heating to 44 degrees C before maximal exercise compared with active nondiabetic individuals, but after an exercise bout, it remains significantly blunted only in diabetic individuals who are sedentary. These findings occur independently of changes in interstitial NO levels. PMID- 12766129 TI - Relation between serum 3-deoxyglucosone and development of diabetic microangiopathy. AB - OBJECTIVE: 3-Deoxyglucosone (3-DG), a highly reactive intermediate of the glycation reaction, has been suggested to contribute to the development of diabetes complications. To verify this hypothesis, we assessed the relation between serum 3-DG concentrations and the severity of diabetic microangiopathy in diabetic patients. RESEARCH DESIGN AND METHODS: We conducted a high-performance liquid chromatography assay to determine the serum 3-DG concentrations of 110 diabetic patients with different degrees of severity of diabetic microangiopathy and 57 age-matched control subjects. RESULTS: The fasting serum 3-DG level in diabetic patients was significantly (P < 0.001) higher than that in control subjects (353 +/- 110 vs. 199 +/- 53 nmol/l). The 3-DG levels were significantly (P < 0.001) elevated even in the diabetic patients showing normoalbuminuria (n = 62, 322 +/- 79 nmol/l) compared with control subjects. The 3-DG levels were further elevated in the patients with microalbuminuria (n = 30, 383 +/- 146 nmol/l) and overt proteinuria (n = 18, 410 +/- 100 nmol/l) (P = 0.027 and P < 0.001 vs. normoalbuminuria group, respectively). This phenomenon was basically reproduced in a category of retinopathy. Furthermore, the diabetic patients with low nerve conduction velocity showed a tendency to display higher 3-DG levels. CONCLUSIONS: The present results show that the fasting serum 3-DG level is elevated in diabetic patients and that the patients with relatively higher 3-DG levels were prone to suffer from more severe complications, indicating a possible association of 3-DG with diabetic microangiopathy. PMID- 12766130 TI - The association between cardiovascular autonomic neuropathy and mortality in individuals with diabetes: a meta-analysis. AB - OBJECTIVE: To examine by meta-analysis the relationship between cardiovascular autonomic neuropathy (CAN) and risk of mortality in individuals with diabetes. RESEARCH DESIGN AND METHODS: We searched Medline for English-language articles published from 1966 to 2001. Fifteen studies having a baseline assessment of cardiovascular autonomic function and mortality follow-up were identified. The analyses were stratified according to whether a single abnormality or two or more measures of cardiovascular autonomic function were used to define CAN. A global measure of association (i.e., relative risk) was generated for each group by pooling estimates across the studies using the Mantel-Haenszel procedure. RESULTS: CAN was significantly associated with subsequent mortality in both groups, although the magnitude of the association was stronger for those studies for which two or more measures were used to define CAN. The pooled relative risk for studies that defined CAN with the presence of two or more abnormalities was 3.45 (95% CI 2.66-4.47; P < 0.001) compared with 1.20 (1.02-1.41; P = 0.03) for studies that used one measure. CONCLUSIONS: These results support an association between CAN and increased risk of mortality. The stronger association observed in studies defining CAN by the presence of two or more abnormalities may be due to more severe autonomic dysfunction in these subjects or a higher frequency of other comorbid complications that contributed to their higher mortality risk. Future studies should evaluate whether early identification of subjects with CAN can lead to a reduction in mortality. PMID- 12766131 TI - Hypoglycemia in diabetes. AB - Iatrogenic hypoglycemia causes recurrent morbidity in most people with type 1 diabetes and many with type 2 diabetes, and it is sometimes fatal. The barrier of hypoglycemia generally precludes maintenance of euglycemia over a lifetime of diabetes and thus precludes full realization of euglycemia's long-term benefits. While the clinical presentation is often characteristic, particularly for the experienced individual with diabetes, the neurogenic and neuroglycopenic symptoms of hypoglycemia are nonspecific and relatively insensitive; therefore, many episodes are not recognized. Hypoglycemia can result from exogenous or endogenous insulin excess alone. However, iatrogenic hypoglycemia is typically the result of the interplay of absolute or relative insulin excess and compromised glucose counterregulation in type 1 and advanced type 2 diabetes. Decrements in insulin, increments in glucagon, and, absent the latter, increments in epinephrine stand high in the hierarchy of redundant glucose counterregulatory factors that normally prevent or rapidly correct hypoglycemia. In insulin-deficient diabetes (exogenous) insulin levels do not decrease as glucose levels fall, and the combination of deficient glucagon and epinephrine responses causes defective glucose counterregulation. Reduced sympathoadrenal responses cause hypoglycemia unawareness. The concept of hypoglycemia-associated autonomic failure in diabetes posits that recent antecedent hypoglycemia causes both defective glucose counterregulation and hypoglycemia unawareness. By shifting glycemic thresholds for the sympathoadrenal (including epinephrine) and the resulting neurogenic responses to lower plasma glucose concentrations, antecedent hypoglycemia leads to a vicious cycle of recurrent hypoglycemia and further impairment of glucose counterregulation. Thus, short-term avoidance of hypoglycemia reverses hypoglycemia unawareness in most affected patients. The clinical approach to minimizing hypoglycemia while improving glycemic control includes 1) addressing the issue, 2) applying the principles of aggressive glycemic therapy, including flexible and individualized drug regimens, and 3) considering the risk factors for iatrogenic hypoglycemia. The latter include factors that result in absolute or relative insulin excess: drug dose, timing, and type; patterns of food ingestion and exercise; interactions with alcohol and other drugs; and altered sensitivity to or clearance of insulin. They also include factors that are clinical surrogates of compromised glucose counterregulation: endogenous insulin deficiency; history of severe hypoglycemia, hypoglycemia unawareness, or both; and aggressive glycemic therapy per se, as evidenced by lower HbA(1c) levels, lower glycemic goals, or both. In a patient with hypoglycemia unawareness (which implies recurrent hypoglycemia) a 2- to 3-week period of scrupulous avoidance of hypoglycemia is advisable. Pending the prevention and cure of diabetes or the development of methods that provide glucose-regulated insulin replacement or secretion, we need to learn to replace insulin in a much more physiological fashion, to prevent, correct, or compensate for compromised glucose counterregulation, or both if we are to achieve near-euglycemia safely in most people with diabetes. PMID- 12766132 TI - Lessons learned: Challenges in interpreting diabetes concepts in the Navajo language. PMID- 12766134 TI - Should postprandial glucose be measured and treated to a particular target? Yes. PMID- 12766133 TI - Lung dysfunction in diabetes. PMID- 12766135 TI - Inflammation and insulin resistance. PMID- 12766136 TI - Weight loss and endothelial function in obesity. PMID- 12766137 TI - Testosterone and atherosclerosis progression in men. PMID- 12766138 TI - Screening for celiac disease in children with type 1 diabetes: two views of the controversy. PMID- 12766140 TI - Status of research funded by the American Diabetes Association--year 4. PMID- 12766139 TI - Validation of interstitial fluid continuous glucose monitoring in cystic fibrosis. PMID- 12766141 TI - The effect of oral glucosamine sulfate on insulin sensitivity in human subjects. PMID- 12766142 TI - Multiple tumors in mitochondrial diabetes associated with tRNALeu(UUR) mutation at position 3264. PMID- 12766145 TI - Physical signs of the intrinsic minus foot. PMID- 12766146 TI - Vietnamese type 2 diabetic subjects with normal BMI but high body fat. PMID- 12766143 TI - Value of a grocery cart and walker in identification and management of symptomatic spinal stenosis in diabetic patients presenting with peripheral neuropathy or claudication. PMID- 12766147 TI - The ability of foot compensation to added weight is reduced in patients with diabetic neuropathy. PMID- 12766151 TI - Adenine release is fast in MutY-catalyzed hydrolysis of G:A and 8-Oxo-G:A DNA mismatches. AB - MutY, a DNA repair enzyme, is unusual in that it binds exceedingly tightly to its products after the chemical steps of catalysis. Until now it was not known whether the product being released in the rate-limiting step was DNA, adenine, or both. MutY hydrolyzes adenine from 8-oxo-G:A (OG:A) base pair mismatches as the first step in the base excision repair pathway, as well as from G:A mismatches. The products are adenine and DNA containing an apurinic (AP) site. Tight product binding may have a physiological role in preventing further damage at the OG:AP site. We developed a rate assay using [8-14C]adenine in OG:A or G:A mismatches that distinguishes between adenine hydrolysis and adenine release. [8-14C]Adenine was released quickly from the MutY.AP-DNA.[8-14C]adenine complex, with a rate constant greater than 5 min-1. This was much faster than the rate-limiting step, at 0.006-0.015 min-1. Gel retardation experiments showed that AP-DNA release was very slow, consistent with it being the rate-limiting step. Thus, the kinetic mechanism involves fast adenine release after hydrolysis followed by rate limiting AP-DNA release. Adenine appears to be buried deep in the protein.DNA interface, but there is enough flexibility or open space for it to dissociate from the MutY.APDNA.adenine complex. These results have implications for the catalytic mechanism of MutY. PMID- 12766153 TI - MiaB protein from Thermotoga maritima. Characterization of an extremely thermophilic tRNA-methylthiotransferase. AB - In Escherichia coli, the MiaB protein catalyzes the methylthiolation of N-6 isopentenyl adenosine in tRNAs, the last reaction step during biosynthesis of 2 methylthio-N-6-isopentenyl adenosine (ms2i6A-37). For the first time the thermophilic bacterium Thermotoga maritima is shown here to contain such a MiaB tRNA-modifying enzyme, named MiaBTm, and to synthesize ms2i6A-37 as demonstrated by an analysis of modified nucleosides from tRNA hydrolysates. The corresponding gene (TM0653) was identified by sequence similarity to the miaB gene cloned and expressed in E. coli. MiaBTm was purified to homogeneity and thoroughly characterized by biochemical and spectroscopic methods. It is a monomer of 443 residues with a molecular mass of 50,710 kilodaltons. Its amino acid sequence shares the CysXXX-CysXXCys sequence with MiaB from E. coli as well as with biotin synthase and lipoate synthase. This sequence was shown to be essential for chelation of an iron-sulfur center and for activity in these enzymes. As isolated, MiaBTm contains both iron and sulfide and an apoprotein form can coordinate up to 4 iron and 4 sulfur atoms per polypeptide chain. UV-visible absorption, resonance Raman, variable temperature magnetic circular dichroism, and EPR spectroscopy of MiaBTm indicate the presence of a [4Fe-4S]+2/+1 cluster under reducing and anaerobic conditions, whereas [3Fe-4S]+1 and [2Fe-2S]+2 forms are generated under aerobic conditions. The redox potential of the [4Fe-4S]+2/+1 transition is -495 +/- 10 mV (versus the normal hydrogen electrode). Finally, the expression of MiaBTm from T. maritima in an E. coli mutant strain lacking functional miaB gene allowed production of ms2i6A-37. These results provide further information on the enzymes involved in methylthiolation of tRNAs. PMID- 12766154 TI - DNA damage-mediated apoptosis induced by selenium compounds. AB - Selenium (Se) compounds, which are the most extensively studied cancer chemopreventive agents, induce apoptotic death of tumor cells. In the current study, we show that selenite-induced apoptosis involves DNA damage. We showed that selenite-induced apoptosis as evidenced by cleavage of poly(ADP-ribose) polymerase was reduced in NIH 3T3 cells treated with ATM small interfering RNA, suggesting the involvement of the DNA damage regulator ATM. Consistent with ATM/ATR involvement, selenite was also shown to stimulate Ser-139 phosphorylation of the ATM/ATR substrate H2AX. Selenite-induced apoptosis was shown to involve DNA topoisomerase II (Top II) as selenite-induced apoptosis was reduced in Top II deficient HL-60/MX2 cells and in HL-60 cells co-treated with the Top II catalytic inhibitor ICRF-193. Using purified human recombinant Top II, selenite was shown to induce reversible Top II cleavage complexes in vitro. In the aggregate, these results suggest that selenite-induced apoptosis, which involves ATM/ATR and Top II, is likely to be because of DNA damage. PMID- 12766152 TI - Human claspin is required for replication checkpoint control. AB - Claspin is a newly identified protein that regulates Chk1 activation in Xenopus. In the present study we investigated the role of human Claspin in the DNA damage/replication checkpoint in mammalian cells. We observed that human Claspin is a cell cycle regulated protein that peaks at S/G2 phase. Claspin localizes in the nuclei, but it only associates with Chk1 following replication stress or other types of DNA damage. In addition, Claspin is phosphorylated in response to replication stress, and this phosphorylation appears to be required for its association with Chk1. Moreover, Claspin interacts with the checkpoint proteins ATR and Rad9. Given that both the ATR and Rad9-Rad1-Hus1 complexes are involved in Chk1 activation, it is possible that Claspin works as an adaptor molecule bringing these molecules together. Using small interfering RNA technology, we have shown that down-regulation of Claspin expression inhibits Chk1 activation in response to replication stress. More importantly, down-regulation of Claspin augments the premature chromatin condensation induced by hydroxyurea, inhibits the UV-induced reduction of DNA synthesis, and decreases cell survival. Taken together, these data imply a potentially critical role for Claspin in replication checkpoint control in mammalian cells. PMID- 12766155 TI - The carboxyl-terminal domain of bacteriophage T7 single-stranded DNA-binding protein modulates DNA binding and interaction with T7 DNA polymerase. AB - Gene 2.5 of bacteriophage T7 is an essential gene that encodes a single-stranded DNA-binding protein (gp2.5). Previous studies have demonstrated that the acidic carboxyl terminus of the protein is essential and that it mediates multiple protein-protein interactions. A screen for lethal mutations in gene 2.5 uncovered a variety of essential amino acids, among which was a single amino acid substitution, F232L, at the carboxyl-terminal residue. gp2.5-F232L exhibits a 3 fold increase in binding affinity for single-stranded DNA and a slightly lower affinity for T7 DNA polymerase when compared with wild type gp2.5. gp2.5-F232L stimulates the activity of T7 DNA polymerase and, in contrast to wild-type gp2.5, promotes strand displacement DNA synthesis by T7 DNA polymerase. A carboxyl terminal truncation of gene 2.5 protein, gp2.5-Delta 26C, binds single-stranded DNA 40-fold more tightly than the wild-type protein and cannot physically interact with T7 DNA polymerase. gp2.5-Delta 26C is inhibitory for DNA synthesis catalyzed by T7 DNA polymerase on single-stranded DNA, and it does not stimulate strand displacement DNA synthesis at high concentration. The biochemical and genetic data support a model in which the carboxyl-terminal tail modulates DNA binding and mediates essential interactions with T7 DNA polymerase. PMID- 12766150 TI - Split dnaE genes encoding multiple novel inteins in Trichodesmium erythraeum. AB - Three inteins were found when analyzing a pair of split dnaE genes encoding the catalytic subunit of DNA polymerase III in the oceanic N2-fixing cyanobacterium Trichodesmium erythraeum. The three inteins (DnaE-1, DnaE-2, and DnaE-3) were clustered in a 70-amino acid (aa) region of the predicted DnaE protein. The DnaE 1 intein is 1258 aa long and three times as large as a typical intein, due to the presence of large tandem repeats in which a 57-aa sequence is repeated 17 times. The DnaE-2 intein has a more typical size of 428 aa with putative protein splicing and endonuclease domains. The DnaE-3 intein is a split intein consisting of a 102-aa N-terminal part and a 36-aa C-terminal part encoded on the first and second split dnaE genes, respectively. Synthesis of a mature DnaE protein is predicted to involve expression of two split dnaE genes followed by two protein cis-splicing reactions and one protein trans-splicing reaction. Tandem repeats in the DnaE-1 intein inhibited the protein splicing activity of this intein when tested in Escherichia coli cells and may potentially regulate DnaE synthesis in vivo. PMID- 12766148 TI - No association between the MTHFR gene polymorphism and diabetic retinopathy in type 2 diabetic patients without overt nephropathy. PMID- 12766156 TI - Mutational analysis of bacteriophage T4 RNA ligase 1. Different functional groups are required for the nucleotidyl transfer and phosphodiester bond formation steps of the ligation reaction. AB - T4 RNA ligase 1 (Rnl1) exemplifies an ATP-dependent RNA ligase family that includes fungal tRNA ligase (Trl1) and a putative baculovirus RNA ligase. Rnl1 acts via a covalent enzyme-AMP intermediate generated by attack of Lys-99 N zeta on the alpha phosphorus of ATP. Mutation of Lys-99 abolishes ligase activity. Here we tested the effects of alanine mutations at 19 conserved positions in Rnl1 and thereby identified 9 new residues essential for ligase activity: Arg-54, Lys 75, Phe-77, Gly-102, Lys-119, Glu-227, Gly-228, Lys-240, and Lys-242. Seven of the essential residues are located within counterparts of conserved nucleotidyltransferase motifs I (99KEDG102), Ia (118SK119), IV (227EGYVA231), and V (238HFKIK242) that comprise the active sites of DNA ligases, RNA capping enzymes, and T4 RNA ligase 2. Three other essential residues, Arg-54, Lys-75 and Phe-77, are located upstream of the AMP attachment site within a conserved domain unique to the Rnl1-like ligase family. We infer a shared evolutionary history and active site architecture in Rnl1 (a tRNA repair enzyme) and Trl1 (a tRNA splicing enzyme). We determined structure-activity relationships via conservative substitutions and examined mutational effects on the isolated steps of Rnl1 adenylylation (step 1) and phosphodiester bond formation (step 3). Lys-75, Lys 240, and Lys-242 were found to be essential for step 1 and overall ligation of 5' phosphorylated RNA but not for phosphodiester bond formation. These results suggest that the composition of the Rnl1 active site is different during steps 1 and 3. Mutations at Arg-54 and Lys-119 abolished the overall RNA ligation reaction without affecting steps 1 and 3. Arg-54 and Lys-119 are thereby implicated as specific catalysts of the RNA adenylation reaction (step 2) of the ligation pathway. PMID- 12766158 TI - Asp274 and his346 are essential for heme binding and catalytic function of human indoleamine 2,3-dioxygenase. AB - L-Tryptophan is the least abundant essential amino acid in humans. Indoleamine 2,3-dioxgyenase (IDO) is a cytosolic heme protein which, together with the hepatic enzyme tryptophan 2,3-dioxygenase, catalyzes the first and rate-limiting step in the major pathway of tryptophan metabolism, the kynurenine pathway. The physiological role of IDO is not fully understood but is of great interest, because IDO is widely distributed in human tissues, can be up-regulated via cytokines such as interferon-gamma, and can thereby modulate the levels of tryptophan, which is vital for cell growth. To identify which amino acid residues are important in substrate or heme binding in IDO, site-directed mutagenesis of conserved residues in the IDO gene was undertaken. Because it had been proposed that a histidine residue might be the proximal heme ligand in IDO, mutation to alanine of the three highly conserved histidines His16, His303, and His346 was conducted. Of these, only His346 was shown to be essential for heme binding, indicating that this histidine residue may be the proximal ligand and suggesting that neither His303 nor His16 act as the proximal ligand. Site-directed mutagenesis of Asp274 also compromised the ability of IDO to bind heme. This observation indicates that Asp274 may coordinate to heme directly as the distal ligand or is essential in maintaining the conformation of the heme pocket. PMID- 12766157 TI - Leukocyte-endothelium interaction promotes SDF-1-dependent polarization of CXCR4. AB - Chemokine-driven migration is accompanied by polarization of the cell body and of the intracellular signaling machinery. The extent to which chemokine receptors polarize during chemotaxis is currently unclear. To analyze the distribution of the chemokine receptor CXCR4 during SDF-1 (CXCL12)-induced chemotaxis, we retrovirally expressed a CXCR4-GFP fusion protein in the CXCR4-deficient human hematopoietic progenitor cell line KG1a. This KG1a CXCR4-GFP cell line showed full restoration of SDF-1 responsiveness in assays detecting activation of ERK1/2 phosphorylation, actin polymerization, adhesion to endothelium under conditions of physiological flow, and (transendothelial) chemotaxis. When adhered to cytokine-activated endothelium in the absence of SDF-1, CXCR4 did not localize to the leading edge of the cell but was uniformly distributed over the plasma membrane. In contrast, when SDF-1 was immobilized on cytokine-activated endothelium, the CXCR4-GFP receptors that were present on the cell surface markedly redistributed to the leading edge of migrating cells. In addition, CXCR4 GFP co-localized with lipid rafts in the leading edge of SDF-1-stimulated cells, at the sites of contact with the endothelial surface. Inhibition of lipid raft formation prevents SDF-1-dependent migration, internalization of CXCR4, and polarization to the leading edge of CXCR4, indicating that CXCR4 surface expression and signaling requires lipid rafts. These data show that SDF-1, immobilized on activated human endothelium, induces polarization of CXCR4 to the leading edge of migrating cells, revealing co-operativity between chemokine and substrate in the control of cell migration. PMID- 12766159 TI - The ATP/substrate stoichiometry of the ATP-binding cassette (ABC) transporter OpuA. AB - ATP-binding cassette (ABC) transport proteins catalyze the translocation of substrates at the expense of hydrolysis of ATP, but the actual ATP/substrate stoichiometry is still controversial. In the osmoregulated ABC transporter (OpuA) from Lactococcus lactis, ATP hydrolysis and substrate translocation are tightly coupled, and the activity of right-side-in and inside-out reconstituted OpuA can be determined accurately. Although the ATP/substrate stoichiometry determined from the uptake of glycine betaine and intravesicular ATP hydrolysis tends to increase with decreasing average size of the liposomes, the data from inside-out reconstituted OpuA indicate that the mechanistic stoichiometry is 2. Moreover, the two orientations of OpuA in proteoliposomes allowed possible contributions from substrate (glycine betaine) inhibition on the trans-side of the membrane and inhibition by ADP to be determined. Here we show that OpuA is not inhibited by up to 400 mm glycine betaine on the trans-side of the membrane. ADP is an inhibitor, but accumulation of ADP was negligible in the assays with inside-out-oriented OpuA, and potential effects of the ATP/ADP ratio on the ATP/substrate stoichiometry determinations could be eliminated. PMID- 12766160 TI - Structural instability and fibrillar aggregation of non-expanded human ataxin-3 revealed under high pressure and temperature. AB - Protein misfolding and formation of structured aggregates are considered to be the earliest events in the development of neurodegenerative diseases, but the mechanism of these biological phenomena remains to be elucidated. Here, we report a study of heat- and pressure-induced unfolding of human Q26 and murine Q6 ataxin 3 using spectroscopic methods. UV absorbance and fluorescence revealed that heat and pressure induced a structural transition of both proteins to a molten globule conformation. The unfolding pathway was partly irreversible and led to a protein conformation where tryptophans were more exposed to water. Furthermore, the use of fluorescent probes (8-anilino-1-naphthalenesulfonate and thioflavin T) allowed the identification of different intermediates during the process of pressure induced unfolding. At high temperature and pressure, human Q26, but not murine Q6, underwent concentration-dependent aggregation. Fourier transform infrared and circular dichroism spectroscopy revealed that these aggregates are characterized by an increased beta-sheet content. As revealed by electron microscopy, heat- and pressure-induced aggregates were different; high temperature treatment led to fibrillar microaggregates (8-10-nm length), whereas high pressure induced oligomeric structures of globular shape (100 nm in diameter), which sometimes aligned to higher order suprastructures. Several intermediate structures were detected in this process. Two factors appear to govern ataxin unfolding and aggregation, the length of the polyglutamine tract and its protein context. PMID- 12766144 TI - Increasing pneumococcal immunizations among people with diabetes using patient reminders. PMID- 12766161 TI - Trichomonas vaginalis Hmp35, a putative pore-forming hydrogenosomal membrane protein, can form a complex in yeast mitochondria. AB - An abundant integral membrane protein, Hmp35, has been isolated from hydrogenosomes of Trichomonas vaginalis. This protein has no known homologue and exists as a stable 300-kDa complex, termed HMP35, in membranes of the hydrogenosome. By using blue native gel electrophoresis, we found the HMP35 complex to be stable in 2 m NaCl and up to 5 m urea. The endogenous Hmp35 protein was largely protease-resistant. The protein has a predominantly beta-sheet structure and predicted transmembrane domains that may form a pore. Interestingly, the protein has a high number of cysteine residues, some of which are arranged in motifs that resemble the RING finger, suggesting that they could be coordinating zinc or another divalent cation. Our data show that Hmp35 forms one intramolecular but no intermolecular disulfide bonds. We have isolated the HMP35 complex by expressing a His-tagged Hmp35 protein in vivo followed by purification with nickel-agarose beads. The purified 300-kDa complex consists of mostly Hmp35 with lesser amounts of 12-, 25-27-, and 32-kDa proteins. The stoichiometry of proteins in the complex indicates that Hmp35 exists as an oligomer. Hmp35 can be targeted heterologously into yeast mitochondria, despite the lack of homology with any yeast protein, demonstrating the compatibility of mitochondrial and hydrogenosomal protein translocation machineries. PMID- 12766162 TI - Glucose metabolism and energy homeostasis in mouse hearts overexpressing dominant negative alpha2 subunit of AMP-activated protein kinase. AB - AMP-activated protein kinase (AMPK) is an energy-sensing enzyme that plays a pivotal role in regulating cellular metabolism for sustaining energy homeostasis under stress conditions. Activation of AMPK has been observed in the heart during acute and chronic stresses, but its functional role has not been completely understood because of the lack of effective activators and inhibitors of this kinase in the heart. We generated transgenic mice (TG) with cardiac-specific overexpression of a dominant negative mutant of the AMPK alpha2 catalytic subunit to clarify the functional role of this kinase in myocardial ischemia. In isolated perfused hearts subjected to a 10-min ischemia, AMPK alpha2 activity in wild type (WT) increased substantially (by 4.5-fold), whereas AMPK alpha2 activity in TG was similar to the level of WT at base line. Basal AMPK alpha1 activity was unchanged in TG and increased normally during ischemia. Ischemia stimulated a 2.5 fold increase in 2-deoxyglucose uptake over base line in WT, whereas the inactivation of AMPK alpha2 in TG significantly blunted this response. Using 31P NMR spectroscopy, we found that ATP depletion was accelerated in TG hearts during no-flow ischemia, and these hearts developed left ventricular dysfunction manifested by an early and more rapid increase in left ventricular end-diastolic pressure. The exacerbated ATP depletion could not be attributed to impaired glycolytic ATP synthesis because TG hearts consumed slightly more glycogen during this period of no-flow ischemia. Thus, AMPK alpha2 is necessary for maintaining myocardial energy homeostasis during ischemia. It is likely that the functional role of AMPK in myocardial energy metabolism resides both in energy supply and utilization. PMID- 12766163 TI - Involvement of a mate chaperone (TorD) in the maturation pathway of molybdoenzyme TorA. AB - As many prokaryotic molybdoenzymes, the trimethylamine oxide reductase (TorA) of Escherichia coli requires the insertion of a bis(molybdopterin guanine dinucleotide)molybdenum cofactor in its catalytic site to be active and translocated to the periplasm. We show in vitro that the purified apo form of TorA was activated weakly when an appropriate bis(molybdopterin guanine dinucleotide)molybdenum source was provided, whereas addition of the TorD chaperone increased apoTorA activation up to 4-fold, allowing maturation of most of the apoprotein. We demonstrate that TorD alone is sufficient for the efficient activation of apoTorA by performing a minimal in vitro assay containing only the components for the cofactor synthesis, apoTorA and TorD. Interestingly, incubation of apoTorA with TorD before cofactor addition led to a significant increase of apoTorA activation, suggesting that TorD acts on apoTorA before cofactor insertion. This result is consistent with the fact that TorD binds to apoTorA and probably modifies its conformation in the absence of cofactor. Therefore, we propose that TorD is involved in the first step of TorA maturation to make it competent to receive the cofactor. PMID- 12766164 TI - Second-site suppressor mutations for the serine 202 to phenylalanine substitution within the interdomain loop of the tetracycline efflux protein Tet(C). AB - The serine 202 to phenylalanine substitution within the cytoplasmic interdomain loop of Tet(C) greatly reduces tetracycline resistance and efflux activity (Saraceni-Richards, C. A., and Levy, S. B. (2000) J. Biol. Chem. 275, 6101-6106). Second-site suppressor mutations were identified following hydroxylamine and nitrosoguanidine mutagenesis. Three mutations, L11F in transmembrane 1 (TM1), A213T in the central interdomain loop, and A270V in cytoplasmic loop 8-9, restored a wild type level of resistance and an active efflux activity in Escherichia coli cells bearing the mutant tet(C) gene. The Tet S202F protein with the additional A270V mutation was expressed in amounts comparable with the original mutant, whereas L11F and A213T Tet(C) protein mutants were overexpressed. Introduction of each single mutation into the wild type tet(C) gene by site-directed mutagenesis did not alter tetracycline resistance or efflux activity. These secondary mutations may restore resistance by promoting a conformational change in the protein to accommodate the S202F mutation. The data demonstrate an interaction of the interdomain loop with other distant regions of the protein and support a role of the interdomain loop in mediating tetracycline resistance. PMID- 12766165 TI - Fluorescence resonance energy transfer analysis of cytochromes P450 2C2 and 2E1 molecular interactions in living cells. AB - The molecular organization of microsomal cytochromes P450 (P450s) and formation of complexes with P450 reductase have been studied previously with isolated proteins and in reconstituted systems. Although these studies demonstrated that some P450s oligomerize in vitro, neither oligomerization nor interactions of P450 with P450 reductase have been studied in living cells. Here we have used fluorescence resonance energy transfer (FRET) to study P450 oligomerization and binding to P450 reductase in live transfected cells. Cytochrome P450 2C2, but not P450 2E1, forms homo-oligomeric structures, and this self-association is mediated by the signal-anchor sequence. Because P450 2C2, in contrast to P450 2E1, is directly retained in the endoplasmic reticulum (ER), these results could suggest that oligomerization may prevent transport from the ER. However, P450 2C1 signal anchor sequence chimera defective in ER retention also formed oligomers, and chimera containing the cytoplasmic domain of P450 2C2, which is directly retained in the ER, did not exhibit self-oligomerization, which indicates that oligomerization is not correlated with direct retention. By using FRET, we have also detected binding of P450 2C2 and P450 2E1 to P450 reductase. In contrast to self-oligomerization, the catalytic domain can mediate an interaction of P450 2C2 with P450 reductase. These results suggest that microsomal P450s may differ in their quaternary structure but that these differences do not detectably affect interaction with the reductase or transport from the ER. PMID- 12766166 TI - Bone morphogenic protein 4 produced in endothelial cells by oscillatory shear stress stimulates an inflammatory response. AB - Atherosclerosis is now viewed as an inflammatory disease occurring preferentially in arterial regions exposed to disturbed flow conditions, including oscillatory shear stress (OS), in branched arteries. In contrast, the arterial regions exposed to laminar shear (LS) are relatively lesion-free. The mechanisms underlying the opposite effects of OS and LS on the inflammatory and atherogenic processes are not clearly understood. Here, through DNA microarrays, protein expression, and functional studies, we identify bone morphogenic protein 4 (BMP4) as a mechanosensitive and pro-inflammatory gene product. Exposing endothelial cells to OS increased BMP4 protein expression, whereas LS decreased it. In addition, we found BMP4 expression only in the selective patches of endothelial cells overlying foam cell lesions in human coronary arteries. The same endothelial patches also expressed higher levels of intercellular cell adhesion molecule-1 (ICAM-1) protein compared with those of non-diseased areas. Functionally, we show that OS and BMP4 induced ICAM-1 expression and monocyte adhesion by a NFkappaB-dependent mechanism. We suggest that BMP4 is a mechanosensitive, inflammatory factor playing a critical role in early steps of atherogenesis in the lesion-prone areas. PMID- 12766167 TI - Prostaglandin E2 stimulates bone sialoprotein (BSP) expression through cAMP and fibroblast growth factor 2 response elements in the proximal promoter of the rat BSP gene. AB - Bone sialoprotein (BSP), an early marker of osteoblast differentiation, has been implicated in the nucleation of hydroxyapatite during de novo bone formation. Prostaglandin E2 (PGE2) has anabolic effects on proliferation and differentiation of osteoblasts via diverse signal transduction systems. Because PGE2 increases the proportion of functional osteoblasts in fetal rat calvarial cell cultures, we investigated the regulation of BSP, as an osteoblastic marker, by PGE2. Treatment of rat osteosarcoma UMR 106 cells with 3 microm, 300 nm, and 30 nm PGE2 increased the steady state levels of BSP mRNA about 2.7-, 2.5-, and 2.4-fold after 12 h. From transient transfection assays, the constructs including the promoter sequence of nucleotides (nt) -116 to +60 (pLUC3) were found to enhance transcriptional activity 3.8- and 2.2-fold treated with 3 microm and 30 nm PGE2 for 12 h. 2-bp mutations were made in an inverted CCAAT box (between nt -50 and 46), a cAMP response element (CRE; between nt -75 and -68), a fibroblast growth factor 2 response element (FRE; nt -92 to -85), and a pituitary-specific transcription factor-1 motif (between nt -111 and -105) within pLUC3 and pLUC7 constructs. Transcriptional stimulation by PGE2 was almost completed abrogated in constructs that included 2-bp mutations in either the CRE and FRE. In gel shift analyses an increased binding of nuclear extract components to double-stranded oligonucleotide probes containing CRE and FRE was observed following treatment with PGE2. These studies show that PGE2 induces BSP transcription in UMR 106 cells through juxtaposed CRE and FRE elements in the proximal promoter of the BSP gene. PMID- 12766168 TI - SLAM-associated protein deficiency causes imbalanced early signal transduction and blocks downstream activation in T cells from X-linked lymphoproliferative disease patients. AB - Deficiency of SAP (SLAM (signaling lymphocyte activation molecule)-associated protein) protein is associated with a severe immunodeficiency, the X-linked lymphoproliferative disease (XLP) characterized by an inappropriate immune reaction against Epstein-Barr virus infection often resulting in a fatal clinical course. Several studies demonstrated altered NK and T cell function in XLP patients; however, the mechanisms underlying XLP disease are still largely unknown. Here, we show that non-transformed T cell lines obtained from XLP patients were defective in several activation events such as IL-2 production, CD25 expression, and homotypic cell aggregation when cells were stimulated via T cell antigen receptor (TCR).CD3 but not when early TCR-dependent events were bypassed by stimulation with phorbol 12-myristate 13-acetate/ionomycin. Analysis of proximal T cell signaling revealed imbalanced TCR.CD3-induced signaling in SAP deficient T cells. Although phospholipase C gamma 1 phosphorylation and calcium response were both enhanced in T cells from XLP patients, phosphorylation of VAV and downstream signal transduction events such as mitogen-activated protein kinase phosphorylation and IL-2 production were diminished. Importantly, reconstitution of SAP expression by retroviral-mediated gene transfer completely restored abnormal signaling events in T cell lines derived from XLP patients. In conclusion, SAP mutation or deletion in XLP patients causes profound defects in T cell activation, resulting in immune deficiency. Moreover, these data provide evidence that SAP functions as an essential integrator in early TCR signal transduction. PMID- 12766169 TI - A novel and highly conserved collagen (pro(alpha)1(XXVII)) with a unique expression pattern and unusual molecular characteristics establishes a new clade within the vertebrate fibrillar collagen family. AB - The type XXVII collagen gene codes for a novel vertebrate fibrillar collagen that is highly conserved in man, mouse, and fish (Fugu rubripes). The pro(alpha)1(XXVII) chain has a domain structure similar to that of the type B clade chains (alpha1(V), alpha3(V), alpha1(XI), and alpha2(XI)). However, compared with other vertebrate fibrillar collagens (types I, II, III, V, and XI), type XXVII collagen has unusual molecular features such as no minor helical domain, a major helical domain that is short and interrupted, and a short chain selection sequence within the NC1 domain. Pro(alpha)1(XXVII) mRNA is 9 kb and expressed by chondrocytes but also by a variety of epithelial cell layers in developing tissues including stomach, lung, gonad, skin, cochlear, and tooth. By Western blotting, type XXVII antisera recognized multiple bands of 240-110 kDa in tissue extracts and collagenous bands of 150-140 kDa in the conditioned medium of the differentiating chondrogenic ATDC5 cell line. Phylogenetic analyses revealed that type XXVII, together with the closely related type XXIV collagen gene, form a new, third clade (type C) within the vertebrate fibrillar collagen family. Furthermore, the exon structure of the type XXVII collagen gene is similar to, but distinct from, those of the genes coding for the type A or B clade pro(alpha) chains. PMID- 12766170 TI - Grb2-independent recruitment of Gab1 requires the C-terminal lobe and structural integrity of the Met receptor kinase domain. AB - The Gab1 docking protein forms a platform for the assembly of a multiprotein signaling complex downstream from receptor tyrosine kinases. In general, recruitment of Gab1 occurs indirectly, via the adapter protein Grb2. In addition, Gab1 interacts with the Met/hepatocyte growth factor receptor in a Grb2 independent manner. This interaction requires a Met binding domain (MBD) in Gab1 and is essential for Met-mediated epithelial morphogenesis. The Gab1 MBD has been proposed to act as a phosphotyrosine binding domain that binds Tyr-1349 in the Met receptor. We show that a 16-amino acid motif within the Gab1 MBD is sufficient for interaction with the Met receptor, suggesting that it is unlikely that the Gab1 MBD forms a structured domain. Alternatively, the structural integrity of the Met receptor, and residues upstream of Tyr-1349 located in the C terminal lobe of the kinase domain, are required for Grb2-independent interaction with the Gab1 MBD. Moreover, the substitution of Tyr-1349 with an acidic residue allows for the recruitment of the Gab1 MBD and for phosphorylation of Gab1. We propose that Gab1 and the Met receptor interact in a novel manner, such that the activated kinase domain of Met and the negative charge of phosphotyrosine 1349 engage the Gab1 MBD as an extended peptide ligand. PMID- 12766171 TI - Anticodon recognition in evolution: switching tRNA specificity of an aminoacyl tRNA synthetase by site-directed peptide transplantation. AB - The highly conserved aspartyl-, asparaginyl-, and lysyl-tRNA synthetases compose one subclass of aminoacyl-tRNA synthetases, called IIb. The three enzymes possess an OB-folded extension at their N terminus. The function of this extension is to specifically recognize the anticodon triplet of the tRNA. Three-dimensional models of bacterial aspartyl- and lysyl-tRNA synthetases complexed to tRNA indicate that a rigid scaffold of amino acid residues along the five beta-strands of the OB-fold accommodates the base U at the center of the anticodon. The binding of the adjacent anticodon bases occurs through interactions with a flexible loop joining strands 4 and 5 (L45). As a result, a switching of the specificity of lysyl-tRNA synthetase from tRNALys (anticodon UUU) toward tRNAAsp (GUC) could be attempted by transplanting the small loop L45 of aspartyl-tRNA synthetase inside lysyl-tRNA synthetase. Upon this transplantation, lysyl-tRNA synthetase loses its capacity to aminoacylate tRNALys. In exchange, the chimeric enzyme acquires the capacity to charge tRNAAsp with lysine. Upon giving the tRNAAsp substrate the discriminator base of tRNALys, the specificity shift is improved. The change of specificity was also established in vivo. Indeed, the transplanted lysyl-tRNA synthetase succeeds in suppressing a missense Lys --> Asp mutation inserted into the beta-lactamase gene. These results functionally establish that sequence variation in a small peptide region of subclass IIb aminoacyl-tRNA synthetases contributes to specification of nucleic acid recognition. Because this peptide element is not part of the core catalytic structure, it may have evolved independently of the active sites of these synthetases. PMID- 12766173 TI - The crystal structure of a cockroach pheromone-binding protein suggests a new ligand binding and release mechanism. AB - Pheromone-binding proteins (PBPs) are small helical proteins found in sensorial organs, particularly in the antennae, of moth and other insect species. They were proposed to solubilize and carry the hydrophobic pheromonal compounds through the antennal lymph to receptors, participating thus in the peri-receptor events of signal transduction. The x-ray structure of Bombyx mori PBP (BmorPBP), from male antennae, revealed a six-helix fold forming a cavity that contains the pheromone bombykol. We have identified a PBP (LmaPBP) from the cockroach Leucophaea maderae in the antennae of the females, the gender attracted by pheromones in this species. Here we report the crystal structure of LmaPBP alone or in complex with a fluorescent reporter (amino-naphthalen sulfonate, ANS) or with a component of the pheromonal blend, 3-hydroxy-butan-2-one. Both compounds bind in the internal cavity of LmaPBP, which is more hydrophilic than BmorPBP cavity. LmaPBP structure ends just after the sixth helix (helix F). BmorPBP structure extends beyond the sixth helix with a stretch of residues elongated at neutral pH and folding as a seventh internalized helix at low pH. These differences between LmaPBP and BmorPBP structures suggest that different binding and release mechanism may be adapted to the hydrophilicity or hydrophobicity of the pheromonal ligand. PMID- 12766172 TI - RhoA interaction with inositol 1,4,5-trisphosphate receptor and transient receptor potential channel-1 regulates Ca2+ entry. Role in signaling increased endothelial permeability. AB - We tested the hypothesis that RhoA, a monomeric GTP-binding protein, induces association of inositol trisphosphate receptor (IP3R) with transient receptor potential channel (TRPC1), and thereby activates store depletion-induced Ca2+ entry in endothelial cells. We showed that RhoA upon activation with thrombin associated with both IP3R and TRPC1. Thrombin also induced translocation of a complex consisting of Rho, IP3R, and TRPC1 to the plasma membrane. IP3R and TRPC1 translocation and association required Rho activation because the response was not seen in C3 transferase (C3)-treated cells. Rho function inhibition using Rho dominant-negative mutant or C3 dampened Ca2+ entry regardless of whether Ca2+ stores were emptied by thrombin, thapsigargin, or inositol trisphosphate. Rho induced association of IP3R with TRPC1 was dependent on actin filament polymerization because latrunculin (which inhibits actin polymerization) prevented both the association and Ca2+ entry. We also showed that thrombin produced a sustained Rho-dependent increase in cytosolic Ca2+ concentration [Ca2+]i in endothelial cells overexpressing TRPC1. We further showed that Rho activated Ca2+ entry via TRPC1 is important in the mechanism of the thrombin induced increase in endothelial permeability. In summary, Rho activation signals interaction of IP3R with TRPC1 at the plasma membrane of endothelial cells, and triggers Ca2+ entry following store depletion and the resultant increase in endothelial permeability. PMID- 12766174 TI - Intestinal sugar absorption is regulated by phosphorylation and turnover of protein kinase C betaII mediated by phosphatidylinositol 3-kinase- and mammalian target of rapamycin-dependent pathways. AB - Stimulation of intestinal fructose absorption by phorbol 12-myristate 13-acetate (PMA) results from rapid insertion of GLUT2 into the brush-border membrane and correlates with protein kinase C (PKC) betaII activation. We have therefore investigated the role of phosphatidylinositol 3 (PI3)-kinase and mammalian target of rapamycin in the regulation of fructose absorption by PKC betaII phosphorylation. In isolated jejunal loops, stimulation of fructose absorption by PMA was inhibited by preperfusion with wortmannin or rapamycin, which blocked GLUT2 activation and insertion into the brush-border membrane. Antibodies to the last 18 and last 10 residues of the C-terminal region of PKC betaII recognized several species differentially in Western blots. Extensive cleavage of native enzyme (80/78 kDa) to a catalytic domain product of 49 kDa occurred. PMA and sugars provoked turnover and degradation of PKC betaII by dephosphorylation to a 42-kDa species, which was converted to polyubiquitylated species detected at 180 and 250+ kDa. PMA increased the level of the PKC betaII 49-kDa species, which correlates with the GLUT2 level; wortmannin and rapamycin blocked these effects of PMA. Rapamycin and wortmannin inhibited PKC betaII turnover. PI3-kinase, PDK 1, and protein kinase B were present in the brush-border membrane, where their levels were increased by PMA and blocked by the inhibitors. We conclude that GLUT2-mediated fructose absorption is regulated through PI3-kinase and mammalian target of rapamycin-dependent pathways, which control phosphorylation of PKC betaII and its substrate-induced turnover and ubiquitin-dependent degradation. These findings suggest possible mechanisms for short term control of intestinal sugar absorption by insulin and amino acids. PMID- 12766175 TI - Connexin-36 contributes to control function of insulin-producing cells. AB - Connexin-36 (Cx36) is a gap junction protein expressed by the insulin-producing beta-cells. We investigated the contribution of this protein in normal beta-cell function by using a viral gene transfer approach to alter Cx36 content in the insulin-producing line of INS-1E cells and rat pancreatic islets. Transcripts for Cx43, Cx45, and Cx36 were detected by reverse transcriptase-PCR in freshly isolated pancreatic islets, whereas only a transcript for Cx36 was detected in INS-1E cells. After infection with a sense viral vector, which induced de novo Cx36 expression in the Cx-defective HeLa cells we used to control the transgene expression, Western blot, immunofluorescence, and freeze-fracture analysis showed a large increase of Cx36 within INS-1E cell membranes. In contrast, after infection with an antisense vector, Cx36 content was decreased by 80%. Glucose induced insulin release and insulin content were decreased, whether infected INS 1E cells expressed Cx36 levels that were largely higher or lower than those observed in wild-type control cells. In both cases, basal insulin secretion was unaffected. Comparable observations on basal secretion and insulin content were made in freshly isolated rat pancreatic islets. The data indicate that large changes in Cx36 alter insulin content and, at least in INS-1E cells, also affect glucose-induced insulin release. PMID- 12766177 TI - Vasoconstriction in active skeletal muscles: a potential role for P2X purinergic receptors? AB - There is evidence that ATP acts as a neurotransmitter in vascular smooth muscle and is coreleased with norepinephrine from sympathetic nerves. We hypothesized that P2X-receptor stimulation with the selective P2X-receptor agonist alpha,beta methylene ATP would produce vasoconstriction in resting and exercising skeletal muscle. Six mongrel dogs were instrumented chronically with flow probes on the external iliac arteries of both hindlimbs and a catheter in one femoral artery. The selective P2X agonist alpha,beta-methylene ATP was infused as a bolus into the femoral artery catheter at rest and during mild, moderate, and heavy exercise. Intra-arterial infusions of alpha,beta-methylene ATP elicited reductions in vascular conductance of 54 +/- 5, 49 +/- 8, 39 +/- 8, and 30 +/- 6% at rest, 3 miles/h, 6 miles/h, and 6 miles/h at a 10% grade, respectively. The agonist infusions did not affect blood flow in the contralateral iliac artery. To examine whether nitric oxide is responsible for the attenuated vasoconstrictor response to P2X stimulation, the infusions were repeated in the presence of NG nitro-l-arginine methyl ester. After nitric oxide synthase blockade, intra arterial infusions of alpha,beta-methylene ATP elicited reductions in vascular conductance of 56 +/- 7, 61 +/- 8, 52 +/- 9, and 40 +/- 7% at rest, 3 miles/h, 6 miles/h, and 6 miles/h at a 10% grade, respectively. P2X-receptor responsiveness was attenuated during exercise compared with rest. Blockade of nitric oxide production did not affect the attenuation of P2X-receptor responsiveness during exercise. These data support the hypothesis that P2X purinergic receptors can produce vasoconstriction in exercising skeletal muscle. PMID- 12766179 TI - Effect of high-intensity training on exercise-induced gene expression specific to ion homeostasis and metabolism. AB - Changes in gene expression during recovery from high-intensity, intermittent, one legged exercise were studied before and after 5.5 wk of training. Genes related to metabolism, as well as Na+, K+, and pH homeostasis, were selected for analyses. After the same work was performed before and after the training period, several muscle biopsies were obtained from vastus lateralis muscle. In the untrained state, the Na+-K+-ATPase alpha1-subunit mRNA level was approximately threefold higher (P < 0.01) at 0, 1, and 3 h after exercise, relative to the preexercise resting level. After 3-5 h of recovery in the untrained state, pyruvate dehydrogenase kinase 4 and hexokinase II mRNA levels were elevated 13 fold (P < 0.001) and 6-fold (P < 0.01), respectively. However, after the training period, only pyruvate dehydrogenase kinase 4 mRNA levels were elevated (P < 0.05) during the recovery period. No changes in resting mRNA levels were observed as a result of training. In conclusion, cellular adaptations to high-intensity exercise training may, in part, be induced by transcriptional regulation. After training, the transcriptional response to an exercise bout at a given workload is diminished. PMID- 12766181 TI - Changes in muscle fiber conduction velocity indicate recruitment of distinct motor unit populations. AB - To obtain more insight into the changes in mean muscle fiber conduction velocity (MFCV) during sustained isometric exercise at relatively low contraction levels, we performed an in-depth study of the human tibialis anterior muscle by using multichannel surface electromyogram. The results show an increase in MFCV after an initial decrease of MFCV at 30 or 40% maximum voluntary contraction in all of the five subjects studied. With a peak velocity analysis, we calculated the distribution of conduction velocities of action potentials in the bipolar electromyogram signal. It shows two populations of peak velocities occurring simultaneously halfway through the exercise. The MFCV pattern implies the recruitment of two different populations of motor units. Because of the lowering of MFCV of the first activated population of motor units, the newly recruited second population of motor units becomes visible. It is most likely that the MFCV pattern can be ascribed to the fatiguing of already recruited predominantly type I motor units, followed by the recruitment of fresh, predominantly type II, motor units. PMID- 12766180 TI - Psoas muscle attenuation measurement with computed tomography indicates intramuscular fat accumulation in patients with the HIV-lipodystrophy syndrome. AB - The human immunodeficiency virus (HIV)-lipodystrophy syndrome is characterized by abnormalities of lipid metabolism, glucose homeostasis, and fat distribution. Overaccumulation of intramuscular lipid may contribute to insulin resistance in this population. We examined 63 men: HIV positive with lipodystrophy (n = 22), HIV positive without lipodystrophy (n = 20), and age- and body mass index-matched HIV-negative controls (n = 21). Single-slice computed tomography was used to determine psoas muscle attenuation and visceral fat area. Plasma free fatty acids (FFA), lipid profile, and markers of glucose homeostasis were measured. Muscle attenuation was significantly decreased in subjects with lipodystrophy [median (interquartile range), 55.0 (51.0-58.3)] compared with subjects without lipodystrophy [57.0 (55.0-59.0); P = 0.05] and HIV-negative controls [59.5 (57.3 64.8); P < 0.01]. Among HIV-infected subjects, muscle attenuation correlated significantly with FFA (r = -0.38; P = 0.02), visceral fat (r = -0.49; P = 0.002), glucose (r = -0.38; P = 0.02) and insulin (r = -0.60; P = 0.0001) response to a 75-g oral glucose tolerance test. In forward stepwise regression analysis with psoas attenuation as the dependent variable, visceral fat (P = 0.02) and FFA (P < 0.05), but neither body mass index, subcutaneous fat, nor antiretroviral use, were strong independent predictors of muscle attenuation (r2 = 0.39 for model). Muscle attenuation (P = 0.02) and visceral fat (P = 0.02), but not BMI, subcutaneous fat, FFA, or antiretroviral use, were strong independent predictors of insulin response (area under the curve) to glucose challenge (r2 = 0.47 for model). These data demonstrate that decreased psoas muscle attenuation due to intramuscular fat accumulation may contribute significantly to hyperinsulinemia and insulin resistance in HIV-lipodystrophy patients. Further studies are needed to assess the mechanisms and consequences of intramuscular lipid accumulation in HIV-infected patients. PMID- 12766182 TI - Short-term training attenuates muscle TCA cycle expansion during exercise in women. AB - Muscle glycogenolytic flux and lactate accumulation during exercise are lower after 3-7 days of "short-term" aerobic training (STT) in men (e.g., Green HJ, Helyar R, Ball-Burnett M, Kowalchuk N, Symon S, and Farrance B. J Appl Physiol 72: 484-491, 1992). We hypothesized that 5 days of STT would attenuate pyruvate production and the increase in muscle tricarboxylic acid cycle intermediates (TCAI) during exercise, because of reduced flux through the reaction catalyzed by alanine aminotransferase (AAT; pyruvate + glutamate <--> 2-oxoglutarate + alanine). Eight women [22 +/- 1 yr, peak oxygen uptake (Vo2 peak) = 40.3 +/- 4.6 ml. kg-1. min-1] performed seven 45-min bouts of cycle exercise at 70% Vo2 peak over 9 days (1 bout/day; rest only on days 2 and 8). During the first and last bouts, biopsies (vastus lateralis) were obtained at rest and after 5 and 45 min of exercise. Muscle glycogen concentration was approximately 50% higher at rest after STT (493 +/- 38 vs. 330 +/- 20 mmol/kg dry wt; P 6 h in vivo, which is slower than in vitro (<2-3 h). PMID- 12766249 TI - Impulse propagation over tactile and kinaesthetic sensory axons to central target neurones of the cuneate nucleus in cat. AB - Paired, simultaneous recordings were made in anaesthetized cats from the peripheral and central axons of individual tactile and kinaesthetic sensory fibres. The aim was to determine whether failure of spike propagation occurred at any of the three major axonal branch points in the path to their cuneate target neurones, and whether propagation failure may contribute, along with synaptic transmission failures, to limitations in transmission security observed for the cuneate synaptic relay. No evidence for propagation failure was found at the two major axonal branch points prior to the cuneate nucleus, namely, the T-junction at the dorsal root ganglion, and the major branch point near the cord entry point, even for the highest impulse rates (approximately 400 impulses s(-1)) at which these fibres could be driven. However, at the highest impulse rates there was evidence at the central, intra-cuneate recording site of switching between two states in the terminal axonal spike configuration. This appears to reflect a sporadic propagation failure into one of the terminal branches of the sensory axon. In conclusion, it appears that central impulse propagation over group II sensory axons occurs with complete security through branch points within the dorsal root ganglion and at the spinal cord entry zone. However, at high rates of afferent drive, terminal axonal propagation failure may contribute to the observed decline in transmission security within the cuneate synaptic relay. PMID- 12766252 TI - The endogenous cannabinoid anandamide inhibits alpha7 nicotinic acetylcholine receptor-mediated responses in Xenopus oocytes. AB - The effect of the endogenous cannabinoid ligand anandamide on the function of the cloned alpha7 subunit of the nicotinic acetylcholine (ACh) receptor expressed in Xenopus oocytes was investigated by using the two-electrode voltage-clamp technique. Anandamide reversibly inhibited nicotine (10 microM) induced-currents in a concentration-dependent manner (10 nM to 30 microM), with an IC50 value of 229.7 +/- 20.4 nM. The effect of anandamide was neither dependent on the membrane potential nor meditated by endogenous Ca2+ dependent Cl- channels since it was unaffected by intracellularly injected BAPTA and perfusion with Ca2+-free bathing solution containing 2 mM Ba2+. Anandamide decreased the maximal nicotine-induced responses without significantly affecting its potency, indicating that it acts as a noncompetitive antagonist on nicotinic acetylcholine (nACh) alpha7 receptors. This effect was not mediated by CB1 or CB2 receptors, as neither the selective CB1 receptor antagonist N-(piperidin-1-yl)-5-(4-chlorophenyl)-1-(2,4 dichlorophenyl)-4-methyl-1H-pyrazole-3-carboximide hydrochloride (SR 141716A) nor CB2 receptor antagonist N-((1S)-endo-1,3,3-trimethyl-bicyclo-heptan-2-yl]-5-(4 chloro-3-methylphenyl)-1-(4-methylbenzyl)-pyrazole-3-carboxamide (SR 144528) reduced the inhibition by anandamide. In addition, inhibition of nicotinic responses by anandamide was not sensitive to either pertussis toxin treatment or to the membrane permeable cAMP analog 8-Br-cAMP (0.2 mM). Inhibitors of enzymes involved in anandamide metabolism including phenylmethylsulfonyl fluoride, superoxide dismutase, and indomethacin, or the anandamide transport inhibitor AM404 did not prevent anandamide inhibition of nicotinic responses, suggesting that anandamide itself acted on nicotinic receptors. In conclusion, these results demonstrate that the endogenous cannabinoid anandamide inhibits the function of nACh alpha7 receptors expressed in Xenopus oocytes in a cannabinoid receptor independent and noncompetitive manner. PMID- 12766254 TI - Relative efficacy of buprenorphine, nalbuphine and morphine in opioid-treated rhesus monkeys discriminating naltrexone. AB - Efficacy is one determinant of whether a drug is an agonist or an antagonist under a particular set of conditions. Relative efficacy among the micro opioid receptor (MOR) ligands buprenorphine, nalbuphine, and morphine was examined in monkeys dependent on morphine (3.2 mg/kg/day) or l-alpha-acetylmethadol (LAAM) (1.0 mg/kg twice daily) and that discriminated naltrexone (0.0178 mg/kg) from saline. In morphine-treated monkeys, buprenorphine and not nalbuphine substituted for naltrexone. When administered before naltrexone in morphine-treated monkeys, morphine and nalbuphine shifted the naltrexone dose-effect curve to the right, while buprenorphine shifted the naltrexone dose-effect curve to the left. Under conditions of acute morphine deprivation, naltrexone-lever responding was slightly attenuated by buprenorphine and markedly attenuated by nalbuphine and morphine. In LAAM-treated monkeys, buprenorphine substituted completely for naltrexone in only one monkey, while nalbuphine and morphine failed to substitute in any monkey. When administered before naltrexone in LAAM-treated monkeys, buprenorphine, nalbuphine, and morphine dose dependently shifted the naltrexone dose-effect curve to the right, with the exception of one monkey in which buprenorphine shifted the naltrexone dose-effect curve to the left. These results demonstrate that a low efficacy MOR ligand can exert agonist or antagonist actions in the same animal depending on immediate pharmacologic history. The qualitatively different effects of buprenorphine in morphine- and LAAM-treated monkeys might be related to magnitude of dependence insofar as dependence can determine the efficacy required for agonist activity. Thus, buprenorphine has markedly different effects across different levels of opioid dependence. PMID- 12766256 TI - Differential actions of fipronil and dieldrin insecticides on GABA-gated chloride channels in cockroach neurons. AB - Fipronil and dieldrin are known to inhibit GABA receptors in both mammals and insects. However, the mechanism of selective toxicity of these insecticides between mammals and insects remains to be seen. One possible mechanism is that insect GABA receptors are more sensitive than mammalian GABAA receptors to fipronil and dieldrin. We examined differential actions of fipronil and dieldrin on GABA-gated chloride channels in insects and compared them with the data on mammalian GABAA receptors. Neurons were acutely dissociated from the American cockroach thoracic ganglia, and currents evoked by GABA were recorded by the whole-cell patch-clamp technique. GABA-evoked currents were carried by chloride ions, blocked by picrotoxinin, but not by bicuculline. Fipronil inhibited GABA currents with an IC50 value of 28 nM, whereas dieldrin exhibited a dual action potentiation with an EC50 value of 4 nM followed by inhibition with an IC50 value of 16 nM. Fipronil and dieldrin acted on the resting receptor at comparable rates, whereas fipronil blocked the activated receptor 10 times faster than dieldrin. Fipronil inhibition was partially reversible, whereas dieldrin inhibition was irreversible. Fipronil was 59 times more potent on cockroach GABA receptors than on rat GABAA receptors. However, the potentiating and inhibitory potencies of dieldrin in cockroach GABA receptors were comparable with those in rat GABAA receptors. It was concluded that the higher toxicity of fipronil in insects than in mammals is due partially to the higher sensitivity of GABA receptors. The mechanism of dieldrin's selective toxicity must lie in factors other than the sensitivity of GABA receptors. PMID- 12766253 TI - Avasimibe induces CYP3A4 and multiple drug resistance protein 1 gene expression through activation of the pregnane X receptor. AB - In vitro and clinical studies were conducted to characterize the potential of avasimibe, an acyl-CoA/cholesterol acyltransferase inhibitor to cause drug-drug interactions. Clinically, 3- and 6-fold increases in midazolam (CYP3A4 substrate) oral clearance were observed after 50 and 750 mg of avasimibe daily for 7 days, respectively. A 40% decrease in digoxin (P-glycoprotein substrate) area under the curve was observed with 750 mg of avasimibe daily for 10 days. In vitro studies were conducted to define the mechanisms of these interactions. Induction was observed in CYP3A4 activity and immunoreactive protein (EC50 of 200-400 nM) in primary human hepatocytes treated with avasimibe. Rifampin treatment yielded similar results. Microarray analysis revealed avasimibe (1 microM) increased CYP3A4 mRNA 20-fold, compared with a 23-fold increase with 50 microM rifampin. Avasimibe induced P-glycoprotein mRNA by about 2-fold and immunoreactive protein in a dose-dependent manner. Transient transfection assays showed that avasimibe is a potent activator of the human pregnane X receptor (hPXR) and more active than rifampin on an equimolar basis. Drug-drug interaction studies for CYP3A4 using pooled human hepatic microsomes and avasimibe at various concentrations, revealed IC50 values of 20.7, 1.6, and 3.1 microM using testosterone, midazolam, and felodipine as probe substrates, respectively. Our results indicate that avasimibe causes clinically significant drug-drug interactions through direct activation of hPXR and the subsequent induction of its target genes CYP3A4 and multiple drug resistance protein 1. PMID- 12766255 TI - N-n-alkylpyridinium analogs, a novel class of nicotinic receptor antagonists: selective inhibition of nicotine-evoked [3H] dopamine overflow from superfused rat striatal slices. AB - Structural simplification of N-n-alkylnicotinium analogs, antagonists at neuronal nicotinic acetylcholine receptors (nAChRs), was achieved by removal of the N methylpyrrolidino moiety affording N-n-alkylpyridinium analogs with carbon chain lengths of C1 to C20. N-n-Alkylpyridinium analog inhibition of [3H]nicotine and [3H]methyllycaconitine binding to rat brain membranes assessed interaction with alpha4beta2* and alpha7* nAChRs, respectively, whereas inhibition of nicotine evoked 3H overflow from [3H]dopamine ([3H]DA)-preloaded rat striatal slices assessed antagonist action at nAChR subtypes mediating nicotine-evoked DA release. No inhibition of [3H]methyllycaconitine binding was observed, although N n-alkylpyridinium analogs had low affinity for [3H]nicotine binding sites, i.e., 1 to 3 orders of magnitude lower than that of the respective N-n-alkylnicotinium analogs. These results indicate that the N-methylpyrrolidino moiety in the N-n alkylnicotinium analogs is a structural requirement for potent inhibition of alpha4beta2* nAChRs. Importantly, N-n-alkylpyridinium analogs with n-alkyl chains < C10 did not inhibit nicotine-evoked [3H]DA overflow, whereas analogs with n alkyl chains ranging from C10 to C20 potently and completely inhibited nicotine evoked [3H]DA overflow (IC50 = 0.12-0.49 microM), with the exceptions of N-n pentadecylpyridinium bromide (C15) and N-n-eicosylpyridinium bromide (C20), which exhibited maximal inhibition of approximately 50%. The mechanism of inhibition of a representative analog of this structural series, N-n-dodecylpyridinium iodide, was determined by Schild analysis. Linear Schild regression with slope not different from unity indicated competitive antagonism at nAChRs mediating nicotine-evoked [3H]DA overflow and a KB value of 0.17 microM. Thus, the simplified N-n-alkylpyridinium analogs are potent, selective, and competitive antagonists of nAChRs mediating nicotine-evoked [3H]DA overflow, indicating that the N-methylpyrrolidino moiety is not a structural requirement for interaction with nAChR subtypes mediating nicotine-evoked DA release. PMID- 12766258 TI - The orally available spleen tyrosine kinase inhibitor 2-[7-(3,4-dimethoxyphenyl) imidazo[1,2-c]pyrimidin-5-ylamino]nicotinamide dihydrochloride (BAY 61-3606) blocks antigen-induced airway inflammation in rodents. AB - Spleen tyrosine kinase (Syk) tyrosine kinase plays essential roles in receptors for Fc portion of immunoglobulins and B cell receptor complex signaling in various inflammatory cells; therefore, inhibitors of Syk kinase may show potential as antiasthmatic/allergic therapeutics. We identified 2-[7-(3,4 dimethoxyphenyl)-imidazo[1,2-c]pyrimidin-5-ylamino]-nicotinamide dihydrochloride (BAY 61-3606), a potent (Ki = 7.5 nM) and selective inhibitor of Syk kinase. BAY 61-3606 inhibited not only degranulation (IC50 values between 5 and 46 nM) but also lipid mediator and cytokine synthesis in mast cells. BAY 61-3606 was highly efficacious in basophils obtained from healthy human subjects (IC50 = 10 nM) and seems to be at least as potent in basophils obtained from atopic (high serum IgE) subjects (IC50 = 8.1 nM). B cell receptor activation and receptors for Fc portion of IgG signaling in eosinophils and monocytes were also potently suppressed by BAY 61-3606. Oral administration of BAY 61-3606 to rats significantly suppressed antigen-induced passive cutaneous anaphylactic reaction, bronchoconstriction, and bronchial edema at 3 mg/kg. Furthermore, BAY 61-3606 attenuated antigen-induced airway inflammation in rats. Based on these anti-inflammatory effects of BAY 61 3606 both in vitro and in vivo, it was demonstrated that Syk may play a very critical role in the pathogenesis of allergic reactions. PMID- 12766251 TI - A small molecule alpha4beta1/alpha4beta7 antagonist differentiates between the low-affinity states of alpha4beta1 and alpha4beta7: characterization of divalent cation dependence. AB - An alpha4beta1/alpha4beta7 dual antagonist, 35S-compound 1, was used as a model ligand to study the effect of divalent cations on the activation state and ligand binding properties of alpha4 integrins. In the presence of 1 mM each Ca2+/Mg2+, 35S-compound 1 bound to several cell lines expressing both alpha4beta1 and alpha4beta7, but 2S-[(1-benzenesulfonyl-pyrrolidine-2S-carbonyl)-amino]-4-[4 methyl-2S-(methyl-[2-[4-(3-o-tolyl-ureido)-phenyl]-acetyl]-amino) pentanoylamino] butyric acid (BIO7662), a specific alpha4beta1 antagonist, completely inhibited 35S-compound 1 binding, suggesting that alpha4beta1 was responsible for the observed binding. 35S-Compound 1 bound RPMI-8866 cells expressing predominantly alpha4beta7 with a KD of 1.9 nM in the presence of 1 mM Mn2+, and binding was inhibited only 29% by BIO7662, suggesting that the probe is a potent antagonist of activated alpha4beta7. With Ca2+/Mg2+, 35S-compound 1 bound Jurkat cells expressing primarily alpha4beta1 with a KD of 18 nM. In contrast, the binding of 35S-compound 1 to Mn2+-activated Jurkat cells occurred slowly, reaching equilibrium by 60 min, and failed to dissociate within another 60 min. The ability of four alpha4beta1/alpha4beta7 antagonists to block binding of activated alpha4beta1 or alpha4beta7 to vascular cell adhesion molecule-1 or mucosal addressin cell adhesion molecule-1, respectively, or to 35S-compound 1 was measured, and a similar rank order of potency was observed for native ligand and probe. Inhibition of 35S-compound 1 binding to alpha4beta1 in Ca2+/Mg2+ was used to identify nonselective antagonists among these four. These studies demonstrate that alpha4beta1 and alpha4beta7 have distinct binding properties for the same ligand, and binding parameters are dependent on the state of integrin activation in response to different divalent cations. PMID- 12766257 TI - A selective and oral small molecule inhibitor of vascular epithelial growth factor receptor (VEGFR)-2 and VEGFR-1 inhibits neovascularization and vascular permeability. AB - Vascular endothelial growth factor (VEGF) is a key driver of the neovascularization and vascular permeability that leads to the loss of visual acuity in diabetic retinopathy and neovascular age-related macular degeneration. Our aim was to identify an orally active, selective small molecule kinase inhibitor of vascular endothelial growth factor receptor (VEGFR)-2 with activity against both VEGF-induced angiogenesis and vascular permeability. We used a biochemical assay to identify 3-[5-methyl-2- (2-oxo-1,2-dihydro-indol-3 ylidenemethyl)-1H-pyrrol-3-yl]-proprionic acid (SU10944), a pyrrole indolinone, which is a potent ATP-competitive inhibitor of VEGFR-2 (Ki of 21 +/- 5 nM). In cellular assays, SU10944 inhibited VEGF-induced receptor autophosphorylation (IC50 of 227 +/- 80 nM) as well as downstream signaling (IC50 of 102 +/- 27 nM). In biochemical assays, SU10944 exhibits potent inhibitory activity against VEGFR 1; weak activity against other related subgroup members, including stem cell factor receptor (SCFR), platelet-derived growth factor receptor beta (PDGFRbeta), and fibroblast growth factor receptor-1 (FGFR-1); and no detectable activity against other protein tyrosine kinases such as epidermal growth factor receptor (EGFR), Src, and hepatocyte growth factor receptor. In cellular assays, the selectivity for SU10944 to inhibit VEGFR is maintained compared with other tyrosine kinases (IC50 for SCFR of 1.6 +/- 0.3 microM, for PDGFRbeta of 30.6 +/- 13.3 microM, for FGFR-1 of >50 microM, and for EGFR of >50 microM). Upon oral administration, SU10944 gave a clear dose response in the corneal micropocket model with an ED50 value for inhibition of neovascularization of approximately 30 mg/kg and a maximum inhibition of 95% at 300 mg/kg. Similarly, upon oral administration in the Miles assay, SU10944 potently inhibited VEGF-induced vascular permeability. Our data indicate that small molecule inhibitors of VEGFR signaling have the potential to ameliorate VEGF-induced neovascularization as well as vascular permeability. PMID- 12766259 TI - cDNA microarray analysis reveals a nuclear factor-kappaB-independent regulation of macrophage function by adenosine. AB - Adenosine is released into the extracellular space from nerve terminals and cells subjected to ischemic stress. This nucleoside modulates a plethora of cellular functions via occupancy of specific receptors. Adenosine is also an important endogenous regulator of macrophage function, because it suppresses the production of a number of proinflammatory cytokines by these cells. However, the mechanisms of this anti-inflammatory effect have not been well characterized. We hypothesized that adenosine may exert some of its anti-inflammatory effects by decreasing activation of the transcription factor nuclear factor-kappaB (NF kappaB), because gene expression of most of the proinflammatory cytokines inhibited by adenosine is dependent on NF-kappaB activation. Using bacterial lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages, we found that adenosine as well as adenosine receptor agonists decreased the production of tumor necrosis factor (TNF)-alpha, a typical NF-kappaB-regulated cytokine. This effect of adenosine was not due to an action on the process of TNF-alpha release, because adenosine suppressed also the intracellular levels of TNF-alpha. However, cDNA microarray analysis revealed that mRNA levels of neither TNF-alpha nor other cytokines were altered by adenosine in either LPS-activated or quiescent macrophages. In addition, although LPS induced expression of a number of other, noncytokine genes, including the adenosine A2b receptor, adenosine did not affect the expression of these genes. Furthermore, adenosine as well as adenosine receptor agonists failed to decrease LPS-induced NF-kappaB DNA binding, NF-kappaB promoter activity, p65 nuclear translocation, and inhibitory kappaB degradation. Together, our results suggest that the anti-inflammatory effects of adenosine are independent of NF-kappaB. PMID- 12766261 TI - Effect of transport inhibitors and additional anti-HIV drugs on the movement of lamivudine (3TC) across the guinea pig brain barriers. AB - To treat human immunodeficiency virus (HIV) within the central nervous system (CNS), levels of anti-HIV drugs in the brain must reach therapeutic concentrations. The ability of (-)-2'-deoxy-3'-thiacytidine (3TC; lamivudine) to cross the blood-brain and blood-cerebrospinal fluid (CSF) barriers, alone and in combination with additional nucleoside analogs, was investigated. The bilateral in situ guinea pig brain perfusion method, linked to high-performance liquid chromatography analyses, was used to examine 3TC uptake into brain and CSF simultaneously. The influence of transport inhibitors and additional nucleoside analogs on this uptake was investigated. 3TC movement across the blood-CSF barrier was examined in more detail by the isolated choroid plexus model. 3TC movement across the brain barriers and subsequent accumulation in the brain and CSF was low. However, 3TC uptake from blood into choroid plexus (a potential CNS target for HIV treatment) was significant, and was facilitated by a digoxin sensitive transporter. Another transporter was identified, which removed 3TC from the choroid plexus. Abacavir, 2'3'-didehydro-3'deoxythymidine, and 3'-azido 3' deoxythymidine did not interact with 3TC at either of the brain barriers to affect CNS concentrations of 3TC. However, a significant interaction between 3TC and 2'3'-dideoxyinosine was observed at the choroid plexus, and it may prove beneficial to select drug combinations where no such interaction is indicated. PMID- 12766260 TI - Nicotine induces a long QT phenotype in Kcnq1-deficient mouse hearts. AB - We have previously shown that targeted disruption of the mouse Kcnq1 gene produces a long QT phenotype in vivo that requires extracardiac factors for manifestation (Casimiro et al., 2001). In the present study, we explore the hypothesis that autonomic neuroeffector transmission represents the "extra cardiac" stimulus that induces a long QT phenotype in mouse hearts lacking Kcnq1. Using the isolated perfused (Langendorff) mouse heart preparation, we challenged wild-type (Kcnq1+/+) and mutant (Kcnq1-/-) mouse hearts with nicotine, an autonomic stimulant. ECGs were recorded continuously, and QT intervals were compared at baseline and peak nicotine-induced heart rates. No significant differences in QT or any other ECG parameters were observed in Kcnq1+/+ versus Kcnq1-/- hearts at baseline. In the presence of nicotine, however, the JT, QT, and rate-corrected QT (QTc) intervals were significantly prolonged in Kcnq1-/- hearts relative to Kcnq1+/+ hearts (e.g., QTc = 92 +/- 11 ms versus 66 +/- 2 ms, respectively, p < 0.01). Similar findings were obtained when the hearts were challenged with either epinephrine or isoproterenol (0.1 microM each), thereby suggesting that sympathetic stimulation drives the long QT phenotype in Kcnq1 deficient hearts. This idea is supported by in vivo ECG data obtained from unrestrained conscious mice using radiotelemetry recording techniques. Again, no significant ECG differences were observed in Kcnq1-/- versus Kcnq1+/+ mice at baseline, but handling/injection stress led to significant QTc increases in Kcnq1 /- mice relative to wild-type controls (11 +/- 3 versus -1 +/- 1%, respectively, p < 0.05). These data suggest that sympathetic stimulation induces a long QT phenotype in Kcnq1-deficient mouse hearts. PMID- 12766265 TI - [Present and future of flat panel detectors in the world]. AB - Present status of development of flat panel detectors and their clinical application in the world have been surveyed, and future trends are also explored especially in the field of material researches and methods of manufacturing. Also the importance of role of medical physicists on user side is described because characteristic physics measurement of a detector assembly is unavoidable and essential in quality assurance in clinical routine and acceptance test in hospitals. Even though physics measurements and clinical evaluations on flat panel detectors have shown remarkable progress and advances in these several years, future problems of cost down in manufacturing and quality assurance to prevent individual differences between detector assemblies must be resolved. Results of evaluation in mammography, chest radiography, fluoroscopy for cardiovascular examination, bone tumor examination and radiotherapy application indicate that flat panel detectors are future promising materials. Their systematic operation is contributing to heighten accuracy of image examinations and preciseness of radiation therapy. Encouragement to medical physicists relevant to flat panel detectors is also raised in this paper. PMID- 12766263 TI - Travel associated legionnaires' disease in Europe in 2000 and 2001. AB - The European Surveillance Scheme for Travel Associated Legionnaires' Disease (EWGLINET) was notified of 360 cases in 2000 and 481 cases in 2001, the highest number reported since 1987. This increase reflects enhanced surveillance activities, especially in the Netherlands, France and Italy, mainly through urinary antigen detection test (78% of cases in 2001). The median delay in reporting to the scheme fell to under 30 days, at the cost of some loss of information on the outcomes of illness. In 2000, 28 clusters were detected compared to 72 in 2001, most of this rise resulting from a change in the definition of clusters. In 2000 and 2001, 55 and 140 environmental investigations were reported respectively. PMID- 12766262 TI - The HIV infection in Europe: large East-West disparity. AB - In 2001, western Europe faces an endemic situation for AIDS (22.8 cases per million population) and for HIV infection (54.9 cases per million), the most affected groups remaining injecting drug users and the homo/bisexual men. However, numbers of new HIV diagnoses are increasing among persons infected through heterosexual contact. Central Europe have been relatively spared, with AIDS incidence under 6 cases per million per year, and new HIV diagnoses between 7 and 10 cases per million. On the other hand, eastern Europe shows an epidemic increase in the number of newly diagnosed HIV infections (233 cases in 1994, around 100,000 reported cases in 2001, ie 349 cases per million population) affecting all countries. PMID- 12766266 TI - [Present state and future of flat panel detector in Japan]. AB - In our country, the introduction of flat panel detector is carried out in the diagnosis region since before several years. In flat panel detector with the high image characteristic, large expectation is being placed. In this paper, image characteristic of flat panel detector and effectiveness of clinical application were reviewed. It is anticipated that flat panel detector with the excellent image characteristic gives large effect in the radiological diagnosis. PMID- 12766267 TI - [Flat-panel detector technology -State-of-the-art and future prospects-]. AB - A flat-panel detector (FPD) is a long-awaited technology to implement the digital X-ray imaging technology into the radiological department. This paper describes the state-of-the-art technology and future prospects on the FPD technology. State of-the-art technology was reviewed taking the CXDI series as an example. Several FPD-based systems have been introduced into the Japanese market since CXDI-11 opened it in November 1998. Accompanying CXDI-C2 for control, CXDI-22 for table position and CXDI-31 for portable, the CXDI series fulfills the requirement of the radiography room being a fully digitalized room. The FPD on the CXDI series is comprised of a scintillator (Gd(2)O(2)S:Tb(3+)) as a primary sensor in which the X-ray is captured and an amorphous silicon detector (LANMIT) as a secondary sensor in which the fluorescent light is detected. Since the scintillator is identical to that of the screen-film systems, it can be said as proven, durable and chemically stable and it is expected to produce the same image quality as the screen-film systems. CXDI-31, a portable FPD-based system, was developed targeting thinner dimensions, lightweight, durability and high spatial resolution. Thoroughly re-designing the mechanical structure and reducing the power consumption at the readout IC realized thinner dimensions. Introducing the portable note PC technologies successfully combined lightweight with durability. Improving the sensor process and re-designing the layout made the sensor high resolution without compromising the signal-to-noise ratio. Future prospects were overviewed in the aspect of technology and applications. Sensitivity, spatial resolution, frame rate and portability were described as the upcoming technology. Increasing gain and reducing noise will realize higher sensitivity, especially by adopting the PbI(2), HgI(2) or such photoconductor materials as the primary sensor. Pixelized amplifier will also achieve higher sensitivity. Layered sensor designed such that TFT layer and sensitive layer are constructed separately will decrease the pixel pitch lower than 100 microm. The FPD has been applied in radiography, mammography and angiography. It will expand the applications into low-dose fluoroscopy to replace the X-ray image intensifiers and into cone-beam computer tomography. What the FPD brought was mainly the efficient workflow of the X-ray technologist. However, diagnosis efficiency and patient benefit must be improved further more by combining FPD technology into computer-aided diagnosis, tele-radiography or other IT-based technologies. Such prospect may come true in the near future. PMID- 12766268 TI - [Flat Panel Detector Philips introduced and its system direction]. AB - We introduced digital X-ray diagnostic systems with Flat panel detector both in general X-ray systems and in Angiography systems. Our introduced Flat Panel Detector has the latest technology and has Cesium Iodide (CsI) that absorbs X-ray energy and generates visible light. Detected light signals make digital X-ray images. CsI is the most important material because its absorption rate of X-ray influences the strength of output digital signal. The purpose in this paper is checking that is latest Flat Panel Detector pulls out enough capability CsI has. Especially the thickness of CsI relates to X-ray absorption. X-ray absorption rate depended on the thickness of CsI was calculated by using simulated X-ray model and the future direction of Flat Panel Detector system was discussed. PMID- 12766270 TI - [Physical imaging properties of a flat panel X-ray detector system]. AB - We report the physical imaging properties of a flat panel detector (FPD) designed for radiographic imaging applications (Revolution XQ/i digital chest imaging system, G.E. Medical Systems). The imaging properties of the detector were evaluated through measurements of the characteristic curve, modulation transfer function (MTF), and Wiener spectrum. The digital characteristic curves of the FPD system were measured for the two tube voltages (80 kV and 120 kV). They showed that the correlation between the pixel values of FPD and the incident exposure to the FPD was a linear correlation. The dynamic range of characteristic curves had a range from about 0.003 to 2 microC/kg at the exposure. The presampling MTFs for different tube voltage were almost the same, when we measured the MTFs at 80 kV and 120 kV. And also, there was no significant difference between the MTFs measured with the slit in the direction parallel to the horizontal direction and with the slit in the perpendicular direction. The relative resolution of the FPD system was a 46% higher than that of ST-V imaging plate of FCR (Fuji Computed Radiography) system at 2 cycles/mm. The digital Wiener spectrum of the FPD system was about 1/10 lower than that of the CR system. Thus the FPD system can provide superior imaging performance due to both high resolution and low noise. PMID- 12766269 TI - [Canon's flat-panel detector]. AB - We measured and evaluated digital, pre-sampling and overall imaging properties (characteristic curve, modulation transfer function (MTF), Wiener spectrum (WS), noise equivalent quanta (NEQ) ) for Cannon's flat-panel detector (FPD), Fuji computed radiography (FCR) and screen-film (S/F) systems, respectively. First, the digital and overall characteristic curves of FPD and FCR systems were more wide dynamic range than that of the S/F system. Second, the pre-sampling and overall MTF of FPD system were better than those of FCR system a little at lower spatial frequencies than 0.8 mm(-1), but the overall MTF of FPD and FCR systems were worse than that of S/F system a little at all spatial frequencies. Third, the digital and overall WS of FPD system were similar or better than those of FCR system, but the overall WS of FPD and FCR systems were worse than that of S/F system. Fourth, the pre-sampling and overall NEQ of FPD system were better than those of FCR system a little at lower spatial frequencies than 1.6 mm(-1), but the overall NEQ of FPD and FCR systems were worse than that of S/F system at all spatial frequencies. Comparison of chest phantom images showed that the FPD produced images with quality comparable to or higher than those of the FCR system. From these results, we can expect that the FPD is useful machine by using digital image processing and so on in the radiology department. PMID- 12766272 TI - [Hologic's Flat-Panel Detector]. AB - We measured and evaluated digital, pre-sampling and overall imaging properties (characteristic curve, Modulation Transfer Function (MTF), Wiener spectrum (WS), Noise Equivalent Quanta (NEQ) and Detective Quantum Efficiency (DQE)) for Hologic's direct type and Cannon's indirect type of Flat-Panel Detector (FPD). First, the digital and overall characteristic curves of both types of FPD were more wide dynamic range than that of the S/F system. Second, the pre-sampling and overall MTF of the direct-type FPD system were superior to those of the indirect type FPD system. Third, for identical exposures, the digital and overall WS of the direct-type FPD system were similar or worse than those of the indirect-type FPD system, and for larger exposure, the digital WS of the both types of FPD system were smaller, but the overall WS of the both types of FPD systems were larger. Fourth, the digital and overall NEQ and DQE of the direct-type FPD system were worse than those of the indirect-type FPD system at lower spatial frequencies than 1.75 - 2.0 mm(-1), but were worse at higher spatial frequencies than 1.75 - 2.0 mm(-1). We show radiographs made with the direct type of FPD system. Radiographs of square wave chart show the difference in MTF and contrast of the both types of FPD systems. As the result of evaluation of radiographs of chest phantom in point of noise by radiologists and radiological technologists, the direct type of FPD system needed double or more exposure dose than own standard condition, this dose was same as the indirect-type FPD system. And radiologists evaluated radiographs of human body, spatial resolution was very good, but contrast was much more likely to high at standard parameter. Therefore we have to consider exposure condition and image processing for the direct type of FPD system. PMID- 12766273 TI - Evaluation of video capture equipment for secondary image acquisition in the PACS. AB - There are many cases in which picture archiving and communication systems (PACS) are built with old-type existing modalities with no DICOM output. One of the methods for interfacing them to the PACS is to implement video capture (/ frame grabber) equipment. This equipment takes analog video signal output from medical imaging modalities, and amplitude of the video signal is A/D converted and supplied to the PACS. In this report, we measured and evaluated the accuracy at which this video capture equipment could capture the image. From the physical evaluation, we found the pixel values of an original image and its captured image were almost equal in gray level from 20%-90%. The change in the pixel values of a captured image was +/-3 on average. The change of gray level concentration was acceptable and had an average standard deviation of around 0.63. As for resolution, the degradation was observed at the highest physical level. In a subjective evaluation, the evaluation value of the CT image had a grade of 2.81 on the average (the same quality for a reference image was set to a grade of 3.0). Abnormalities in heads, chests, and abdomens were judged not to influence diagnostic accuracy. Some small differences were seen when comparing captured and reference images, but they are recognized as having no influence on the diagnoses. PMID- 12766264 TI - Prevalence of nosocomial infections in two Latvian hospitals. AB - The first point prevalence survey of the nosocomial infection (NI) rate was conducted in two Latvian hospitals. At the time of the survey 17.5% (226/1291) patients had symptoms or were being treated for infection. The overall prevalence rate was 5.6% (72/1291) for NI and 12.7% for community acquired infections (164/1291). Surgical site infection (SSI) was the most common NI (62%) followed by respiratory tract infection (RTI) (7.5%), and urinary tract infection (UTI) (6.4%). NI rate was higher with increasing age of patients, in intensive care units and surgical wards, and among those who had an intravenous device or urinary catheter. Microbiological investigation yielded positive results in 29% (21/72) of patients with NIs. Antibacterial treatment was given to 22.3% (288/1291) of hospitalised patients and in 62% (182/288) of these, cefazolin was prescribed. Results from this study will be used to plan a national prevalence survey. PMID- 12766275 TI - Methodology development for quantitative optimization of security enhancement in medical information systems -Case study in a PACS and a multi-institutional radiotherapy database-. AB - The target of our study is to establish the methodology for analyzing level of security requirements, for searching suitable security measures and for optimizing security distribution to every portion of medical practice. Quantitative expression must be introduced to our study as possible for the purpose of easy follow up of security procedures and easy evaluation of security outcomes or results. Results of system analysis by fault tree analysis (FTA) clarified that subdivided system elements in detail contribute to much more accurate analysis. Such subdivided composition factors very much depended on behavior of staff, interactive terminal devices, kinds of service, and routes of network. As conclusion, we found the methods to analyze levels of security requirements for each medical information systems employing FTA, basic events for each composition factor and combination of basic events. Methods for searching suitable security measures were found. Namely risk factors for each basic event, number of elements for each composition factor and candidates of security measure elements were found. Method to optimize the security measures for each medical information system was proposed. Namely optimum distribution of risk factors in terms of basic events were figured out, and comparison of them between each medical information systems became possible. PMID- 12766276 TI - Shielding effects of body - shields for 3D PET. AB - We analyzed basic physics characteristics of body-shields which have been considered for screening out radioactivity outside the field of view (OFOV) in positron emission tomography (PET). Phantom experiments were performed with simple rectangular body-shields. A Monte Carlo simulation technique was used to analyze the experimental results and to simulate cases that were not examined experimentally. It was confirmed that the body-shields effectively reduced unwanted radiations from OFOV radioactivity for one of the latest commercial PET scanners, the ECAT EXACT HR(+). The geometrical conditions were the most important factor in determining the shielding effect. The shield thickness should be large enough to keep the shield-scatter component low. The body-shield should be carefully designed to minimize the gap between the body-shield and radioactive distribution as much as possible to maximize the shielding effects, and to be applicable to clinical diagnoses. PMID- 12766271 TI - [An experience of the clinical study with angiography system using a Flat Panel Detector]. AB - We had the chance of clinical studies by new proto-type Digital Subtraction Angiography (DSA) System with the Flat Panel Detector (FPD). That system has two types of detector, FPD and I.I.-CCD camera, and we can compare with image quality in clinical. We measured and discussed about the physical characteristic data of both detectors. Up to physical characteristic evaluation, FPD has good MTF and SNR performance, comparing with I.I.-CCD camera. In clinical evaluation, DSA image of FPD was superior to that of I.I.-CCD camera by visual comparison of medical doctors. Finally we discussed about the feasibility of replacement of detectors. We expect that DSA system will accept FPD in near future. PMID- 12766274 TI - The application of a new predictive technique for lossless compression of medical images. AB - A new predictive technique for lossless compression is applied to CT and MR images in order to achieve high compression in the lossless mode. To accomplish this, a prediction value was obtained by assuming the horizontal and vertical components of each pixel. Wide and narrow predictive coding modes were adaptively selected to achieve an accurate prediction. The performance of the predictive technique was compared with the performance of four representative compression techniques in the lossless mode, using different types of CT and MR image sets with different bit rates. Overall, the proposed predictive technique yielded the best compression, particularly with the lower bit-rate images tested. While the compression results are largely dependent on the images used, the results confirm the strong potential of the proposed predictive technique as a strategy for obtaining high lossless-compression in medical images. PMID- 12766277 TI - Development of a time-oriented data warehouse based on a medical information event model. AB - We designed a new medical information event model and developed a time-oriented data warehouse based on the model. Here, the medical information event in a basic data unit is handled by a medical information system. The timing of decision making and treatment for a patient in the processing of his medical information is sometimes very critical. The time-oriented data warehouse was developed, to provide a search feature on the time axis. Our medical information event model has a unique simple data structure. PC-ORDERING2000 developed by NEC, which used Oracle, had about 600 pages of tables. However, we reduced these 600 complicated data structures to one unique and simple event model. By means of shifting clinical data from the old type order entry system into the new order entry system of the medical information event model, we produced a simple and flexible system, and the easy secondary use of clinical data of patients was realized. Evaluation of our system revealed heightened data retrieval efficiency and shortened response time 1:600 at a terminal, owing to the 1:600 reduction of the number of tables as mentioned above. PMID- 12766279 TI - Dose evaluation and effective dose estimation from multi detector CT. AB - Computed tomography (CT) has evolved remarkably through device improvement and advancement of peripherals, including computers. In 1999, multi detector-row CT (MDCT) appeared and rapid high-speed scanning became possible. However, usefulness of MDCT in actual clinical application cannot be assessed until the exposure doses are assessed appropriately. Since CT examinations need a comparatively high dose, it is necessary to evaluate patient exposure for introduction of MDCT. Patient doses by three types of MDCTs were evaluated for cases of scanning of the chest and abdomen-pelvis. The examination conditions were the same as those in actual clinical examinations. The obtained effective doses were 9.4-28 mSv for the chest examination and 13-28 mSv for the abdomen pelvis. The average surface doses varied between 16-43 mGy for the chest examination and 20-37 mGy for the abdomen-pelvis. The highest surface dose was 57 mGy for the abdomen-pelvis examination. The exposed doses differed according to scanning method and imaging conditions such as tube current, slice thickness and so on. It seemed that there is room for dose reduction by proper adjustment of scan conditions in MDCT examinations. PMID- 12766278 TI - Extended and fading boundaries of today's research areas in medical physics and informatics. PMID- 12766283 TI - [Mammography: consideration about radiation from the point of a radiologist's view]. AB - Mammography is becoming popular rapidly in Japan because it is now admitted as a modality for breast screening. Radiation will not cause any adverse effect if mammography screening is done for women over 50 years old, every other year and by only MLO (medio-lateral oblique) view as this is recommended by the government. Radiation for one view is regulated to be less than 3mGy. Radiation by mammography may become problem if it is done for younger women, by more views (i.e. cranio-caudal, other additional views) and more often especially for follow up. Radiation may be more if the system is not adequate. In Japan mammograms are read by surgeons in many institutions and there are not enough medical physicists in each hospital. Cooperation between medical physicists, radiographers and radiologists will be necessary to achieve good quality mammography with adequate radiation. PMID- 12766281 TI - [Fundamental study on stability of dry-processing imager system DRYPRO 722/SD-P]. AB - Dry-processing leaser imager systems have become popular due to the advantages such as the elimination of the cost and labor associated with the wet chemical processing. In this paper, the stability of a dry-processing imager system Drypro 722/SD-P was studied using SMPTE pattern films processed by a dry-processing imager Drypro 722 and a wet-processing imager Li-8/SRX-502 at three different times of the day over a period of five consecutive working days. The dry system is inferior to the wet system on the stability and the spatial uniformity, so that the problems assess the necessity of QC for Drypro 722 Laser Imagers. The instability of the post-processing dry and wet films is also studied. The dry film is very instable compared with the wet film. Accordingly the wet film must be handled with the care. PMID- 12766288 TI - [CR (Computed Radiography)]. AB - Several recent studies have a questionnaire survey of many hospitals regarding the radiation exposure conditions of X-ray examinations. From the survey result the entrance surface doses (first quartile, median, third quartile and mean values) have calculated by using the Numerical Dose Determination (NDD) method. The calculated result for all types of examinations showed that the median values were lower than the mean values, whereas the median values of the results were lower than those in the NRPB 21 in England. The mean values of the results were lower than those in the NRPB 21 in England. The mean values were lower than IAEA guidance levels. The results have indicated that the medical exposure dose (entrance surface dose) is lower with Computed Radiography (CR) than with the Screen/Film (S/F) system except the case of chest radiography. In the case of chest radiography (adult patients) by CR exposure set-up using a lower voltage and high mAs than the S?F method, the entrance surface doses were 150 % of the median value for the overall examination and 160 % of the median value for orthochromatic S/F systems. A difference in patient dose among hospitals using CR system was also found. Mean entrance surface dose in CR system was 0.12 mGy in a quartile, 0.19 mGy in the middle and 0.27 mGy in the third quartile. Among the hospitals that showed higher doses of third quartile than above mentioned, dose differences of a quartile were distributed 2 to 10 times higher than mean exposure doses. Mori et al. propose guidance levels are lower than IAEA guidance levels of entrance surface doses for examinations carried out in Japanese institutions as the 3rd quartile of the dose distributions. Therefore the quality control of tube voltage, tube current and mAs and a regular measurement of exposure for X-ray units are required. PMID- 12766282 TI - [Mammography -a guidance level and the present situation of mammographic dose-]. AB - Mammography practiced for the detection of breast carcinomas requires an appropriate X-ray equipment which can meet the performance standard. Also it is necessary to meet the requirements for the exposure dose as well as image quality. Factors affecting the exposure dose are X ray equipment, especially target-filter combination, recording systems, compression of breast, structural characteristics of breast and etc. The current guidance for the mammography adopts 3mGy as a dose level for standard breast for a direction which is recommended by IAEA. However, the average glandular dose for the mammography was 1.5mGy according to the image evaluation survey of medical institution as of December, 2001. This figure is almost equal to the result given by the national survey conducted from 1997 to 1998. The exposure reduction depends on the technique of radiological technologists. It is needed for them to optimize affecting factors. When reducing breast thickness by at least 1cm, the exposure does can be reduced by 50%. PMID- 12766284 TI - [X-ray exposure dose control for x-ray CT system]. AB - Shortening scan time of CT scanner system has been evolved and increase in number of CT examinations has also been remarkable. This has been resulted from global recognition of usefulness of the CT examination, contrary to this merit, however, it is important to recognize the risk of x-ray exposure dose. Japan Industry Association of Radiological Systems (JIRA) in which CT manufacturers join has issued the concrete countermeasure and guidance for reduction in x-ray exposure dose in response to the ICRP90 Recommendation. Current CT scanner systems provide the data related to x-ray exposure dose such as CTDI(w) for setting CT scan parameters. To reduce x-ray exposure dose against infant patient, the scan parameters specified to infant patient (CT infant protocol) can be provided. Exposure dose by x-ray CT can be measured by the measurement method corresponding to IEC-60601-2-44 and by using phantom. CTDI measurement is made by CTDI(100) that measures in a range of 100mm for all slice thicknesses, and absorbed radiation dose is converted to that of air. Dose profile is measured by using multiple thermoluminescence dosimeter (TDL) chips. CT exposure dose data including CTDI(100) and Dose profile are well-defined, and Dose Information Guide conforming to IEC-60601-2-44 is provided to user for the purpose of reducing x ray exposure dose. Studies by low dose (2.5 approximately 3mA) simulation for the purpose of reducing x-ray exposure dose in screening CT examination of lung cancer and development of ROI scan to reduce x-ray exposure dose in puncture under CT fluoroscopy have also been conducted. PMID- 12766285 TI - [Medical exposure from computed tomographic examination -a recommendation of dosimetry by film method-]. AB - Already, the 30 year have passed since that the CT scanner was developed by Hunsfield. And now above 16,000 sets are at work. On the other hand, as the improvements of image reconstruction algorithm and variations os scan method, the medical exposure is increasing. For example, in the case of computed radiography, medical exposure is stability by using an auto exposure controller but in the case of CT, there is no unit and usually the exposure condition of CT examination are used that of recommended an exposure condition by maker. So the medical exposure greatly differs according to the operators or the kinds of CT scanner. In addition, there is the close connection between the x-ray intensity and the image quality. Especially, in the case of CT scanner there is no saturation of x ray intensity like the optical density of film screen system because the digital radiographic systems have the wide dynamic range. So the increase in x-ray intensity makes a contribution to improvement of the image quality. But the increase in x-ray intensity makes a contribution to the increase in medical exposure. Therefore it is very important that the CT operators grasp the medical exposure per each CT examination and patient. There are many methods of medical exposure dosimetry on CT examination. But almost all the methods except the film method are impossible to get the medical exposure per each CT examination and patient. So in this paper the film method is recommended. Because the film method is low-priced, simple, easy and immediately shows the medical exposure per each CT examination and patient. Furthermore, it is confirmed that the film method is getting the correct medical exposure because that values gotten by the film method and ionization chamber are almost equivalent. PMID- 12766289 TI - A method for measuring the dose distribution of the radiotherapy domain using the computed radiography system. AB - Knowing the dose distribution in a tissue is as important as being able to measure exposure or absorbed dose in radiotherapy. Therefore, we have developed a measurement method for the dose distribution (CR dosimetry) in the phantom based on the imaging plate (IP) of the computed radiography (CR). The IP was applied for the dose measurement as a dosimeter instead of the film used for film dosimetry. The data from the irradiated IP were processed by a personal computer with 10 bits and were depicted as absorbed dose distributions in the phantom. The image of the dose distribution was obtained from the CR system using the DICOM form. The CR dosimetry is an application of CR system currently employed in medical examinations to dosimetry in radiotherapy. A dose distribution can be easily shown by the Dose Distribution Depiction System we developed this time. Moreover, the measurement method is simpler and a result is obtained more quickly compared with film dosimetry. PMID- 12766287 TI - [Dose reduction by several measures in X-ray angiographic system]. AB - "The radiation protection measures from equipment side, from a maker's point of view."TAKAJI SAITO Medical Solutions Marketing Division, Siemens-Asahi Medical Technologies Ltd. Taking care of limited fluoro and acquisition time during every case is an important prerequisite for dose reduction. Latest technology provides us with a number of means for dose savings regarding patients and staff. Here, the practical techniques of the radiation control function in angiography system are addressed. Last Image Hold (LIH). Collimation without radiation on LIH. Pulsed fluoroscopy at high and low rates. Additional spectral filters in the X ray beam. Display of applied dose during the procedure These technological measures will definitely contribute for dose reduction for interventional procedures. PMID- 12766286 TI - [The potential patient skin injuries from radiologically guided interventional procedure: the present condition and recommendable measure]. AB - Radiologically guided interventional procedures may result in excessive radiation dose for the patients. During the last decade, more than 70 cases of radiation skin injuries have been reported. This may be partly because the potential dangers of X-radiation are not yet well recognized by the physicians, and also by lacking of practical and reliable way to monitor the dose of X-ray radiation at the radiology suite. The author presents a few recommendable techniques to monitor the patient's radiation doses during interventional radiological procedures, which may be helpful in preventing patient's radiation injury. PMID- 12766280 TI - Fast determination method of cerebral metabolic rate images of glucose using dynamic PET data. AB - Measurement of the rate constant parameters of the tracer 18F-FDG, used with positron emission tomography (PET) to determine the cerebral metabolic rate of glucose (CMRGlc), can provide a clear understanding of the physiological processes in the human brain. At present, the methods that are widely used to obtain CMRGlc, such as nonlinear least squares (NLS), first require the reconstruction of a time sequence of images. The reconstruction of these images requires a large amount of computation, especially in 3D Depth-of Interaction PET (DOI-PET), and the nonlinear based methods also require a large amount of computation. In this paper, we propose a fast parametric image reconstruction method for 18F-FDG dynamic PET studies. In our method a deconvolving process is first employed on the time sequential projection data to remove the effect of the measured plasma time activity. The deconvolved terms are integrated over three different time intervals and the parameters for determining CMRGlc can be obtained analytically. Our method requires only three reconstructing processes and reduces the computational demand to estimate CMRGlc. The algorithm performance is evaluated using a digital phantom and a clinical data set and the results show that the proposed method produces images with the same or better quality as the images from the NLS method, with much less computation compared to the NLS method. PMID- 12766290 TI - Development of a Virtual Private Database for a multi-institutional Internet based Radiation Oncology Database overcoming differences in protocols. AB - A multi-institutional Radiation Oncology Greater Area Database (ROGAD) was started in 1991 under the direction of the Japanese Society for Therapeutic Radiology and Oncology (JASTRO). Use of ROGAD was intended to allow reflection of results of data analysis into treatment strategy and treatment planning for individual cases, to provide quality assurance, to maximize the efficacy of radiotherapy, to allow assessment of new technologies or new modalities, and to optimize medical decision making. ROGAD collected 13,448 radiotherapy treatment cases from 325 facilities during the period from 1992 to 2001. In 2000, questionnaires were sent to 725 radiotherapy facilities throughout Japan, to further obtain the situation of the radiation oncology database. Workers at 179 facilities replied that"the protocol of my facility is different from ROGAD protocol and I must send data according to the ROGAD protocol". So, we developed the Virtual Private Database System (VPDS) which is operated as if an oncologist had a database solely owned by his own facility, in spite of actually operating ROGAD. VPDS realizes integration of different plural databases, regardless of differences in entry methods, protocols, definitions and interpretations of contents of clinical data elements between facilities. PMID- 12766291 TI - [X-ray refraction contrast imaging]. PMID- 12766293 TI - [X-ray phase imaging using a X-ray tube with a small focal spot -improvement of image quality in mammography-]. AB - Phase contrast X-ray imaging has been studied intensively using X-rays from synchrotron radiation and micro-focus X-ray tubes. However, these studies have revealed the difficulty of this technique's application to practical medical imaging. We have created a phase contrast imaging technique using a molybdenum X ray tube with a small focal spot size for mammography. We identified the radiographic conditions in phase contrast magnification mammography with a screen film system, where edge effect due to phase contrast overcomes geometrical unsharpness caused by the 0.1mm-focal spot of a molybdenum X-ray tube. The edge enhancement due to phase imaging was observed in an image of a plastic tube, and then geometrical configuration of the X-ray tube, the object and the screen-film system was determined for phase imaging of mammography. In order to investigate a potential for medical application of this method, we conducted evaluation of the images of ACR 156 mammography phantom. We obtained higher scores for phase imaging using high speed screen-film systems without any increase of X-ray dose than the score for contact imaging using a standard speed screen-film system. PMID- 12766292 TI - [Refraction-contrast bone imaging using synchrotron radiation]. AB - The X-ray refraction-contrast imaging using synchrotron radiation with some X-ray energies is successfully performed at B120B2 of SPring-8. The refraction-contrast images of bone samples such as human dried proximal phalanx, wrist, upper cervical vertebrae and sella turcica, and as mouse proximal femur, using the synchrotron X-ray are always better in image contrast and resolution than those of the absorption-contrast images using the synchrotron X-ray and/or the conventional X-ray tube. There is much likeness in the image contrast and resolution of trabeculae bone in the human dried proximal phalanx between X-ray energy of 30 keV at sample-to-film distance of 1m and those of 40, 50 keV at those of 4,5m, respectively. High-energy refraction-constrast imaging with suitable sample-to-film distance could reduce the exposure dose in human imaging. In the refraction-contrast imaging of human wrist, upper cervical vertebrae, sella turcica and mouse proximal femur using the synchrotoron X-ray, we can obtain better image contrast and resolution to correctly extract morphological information for diagnosis corresponding to each of the clinical field than those of the absorption-contrast images. PMID- 12766294 TI - [Phase-contrast x-ray imaging with x-ray interferometer for medical applications]. AB - Phase-contrast x-ray imaging with x-ray interferometer can depict the minute difference within the biological object, and its sensitivity is about 1000 times higher than that of absorption-contrast method. For biomedical use of this technique, a large monolithic x-ray interferometer and 2 crystal interferometer having a field of view with 25 mm x 25 mm is being developed. Phase-contrast x ray CT could reveal detail structures within tumor and surrounded tissue, and the vessel imaging of rat liver is also possible using physiological saline at 17.7 keV x-ray energy. Recently, human breast tissues were imaged at 35 keV and the contrast of image was much better than usual absorption contrast x-ray image obtained at 17.7 keV energy. PMID- 12766295 TI - Heavy ion CT system based on measurement of residual range distribution. AB - The heavy ion CT system is proposed which is based on measurement of the residual range distribution by a fluoroscopy detector consisting of an intensifying screen and a CCD video camera. To investigate the fundamental performance of the proposed system, the spatial and density resolutions of the CT image were evaluated. The heavy ion beam 12C accelerated up to 400 MeV/u by HIMAC was used in this study. A series of projection images of two types of phantoms (PMMA pipes with several sizes and various kinds of rods with different electron densities) were experimentally taken for five values of range shifter thickness at each projection angle. From these images, the residual range distribution was obtained and used to reconstruct CT images of the relative stopping power which were obtained by the filtered back projection method with the Shepp-Logan filter after noise reduction processing by the median filter. As a result, the spatial resolution was found to be less than 2 mm and the relative density resolution around the density of water was less than 0.07. PMID- 12766297 TI - [Review of Recent Medical Ultrasonic Technology] AB - This paper was reviewed in recent progress of medical ultrasonic technology. Obviously in 21 century we face to the second generation of ultrasonic technology in medicine. First of all, B-mode imaging and Doppler color flow mapping have been improved by the digital beam forming technology. This technology leads to high research activity in the field of image processing such as automated tumor segmentation. Harmonic imaging is also newly developed technology. This technique uses harmonic component of echoes while the ultrasound waves are traveling through the nonlinear media such as biological tissues so that high quality images are obtained. Contrast echo method is expected to detect small echo signals from slow and small blood flow. Harmonic power Doppler imaging using micro bubbles is expected to detect early stage of cancer. Three dimensional real time imaging is next generation of ultrasonic image diagnosis system. Two dimensional array is a key technology to realize this imaging. Finally, ultrasonic therapy is described. PMID- 12766296 TI - Defects of a mammography quality control phantom visualized by synchrotron radiation imaging. AB - Synchrotron radiation (SR) imaging of an RMI 156 mammography quality control phantom, serial number 156-15330, revealed some defects which degraded the visibility of calcification specks. SR imaging was performed at SPring-8, in Harima, Japan by using a monochromatic energy of 20 KeV with a field-of-view of 24 X 24 mm. Different kinds of images were obtained by changing sample-to detector distances; absorption images and refraction-enhanced images. Specks were embedded in a wax matrix and were imaged as black in an absorption image. In a refraction-enhanced image, they were imaged as a black region with white margins. Foreign objects with opposite contrast were detected near, or overlapped with, some specks. As they were depicted as white in the absorption images and as white with a black margin in the refraction-enhanced image, it seemed that they had low X-ray attenuation and a low refraction index compared with the surrounding wax. They might presumably be air bubbles. Visibility of specks in an absorption image was seriously interfered with when those object(s) overlapped with specks. This kind of defect may cause a difficulty in meeting quality assurance specifications when a facility inadvertently purchases defective phantoms. PMID- 12766299 TI - [Coded excitation medical ultrasound imaging] AB - Pulse compression techniques were developed mainly for radar systems. And the technology allows a radar to utilize a long pulse to achieve radiated energy, but simultaneously to obtain the range resolution of a short pulse. It accomplishes this by employing frequency or phase modulation to widen the signal bandwidth. Coded excitation is an ultrasound technique which has only been made commercially available in the last 3 years. It improves SNR without loss of resolution by transmitting a long binary phase-encoded pulse sequence and then compressing its echo into a short, high-amplitude pulse on receive. Different types of codes, e.g. Golay, may be used to suppress range side lobes. And a new technique called B-Flow for imaging blood flow also uses advanced digital signal encoding/decoding techniques to provide direct visualization of blood echoes. Coded excitation together with tissue equalization forms the basis for B-Flow, which extends the wideband resolution and high frame rate capabilities of B-mode to flow and tissue imaging. A key aspect of B-Flow is tissue equalization, which uses decoding to preferentially reduce non-moving tissue signals to the level of the flow signal such that both may be displayed simultaneously without machine prioritization and overlay (as in color flow). PMID- 12766298 TI - [Present and future of ultrasonic three-dimensional imaging] AB - Three-dimensional (3-D) imaging has recently been utilized for medical diagnosis alongside of the use of cross-sectional imaging. In X-ray CT and MRI equipment, a computer installed inside executes 3-D image reconstruction with the raw data obtained from the patient. However, in ultrasound equipment, it is necessary for 3-D imaging to use specific technologies that take the inherent characteristics of the ultrasonic echo data into account. In this paper, I broadly divide manners of 3-D image rendering into "direct" methods and "indirect" methods. Direct methods have recently prevailed in the area of ultrasonic 3-D imaging, as these methods are particularly appropriate for processing ultrasound data. In terms of trends, both technologies of segmentation and real-time imaging for the 3-D ultrasound will emerge as the dominant technologies in the future. PMID- 12766300 TI - [Advanced Dynamic Flow] AB - There are problems for contrast imaging using LevovistTM contrast agent. In case of Doppler mode, poor resolution and large blooming area are the problems. In case of 2nd harmonic imaging and pulse inversion imaging, tissue harmonic image(THI) becomes the problem because Levovist needs high mechanical index(MI). THI increases under the high MI condition and THI interferes with the contrast image of Levovist. And also bubbles of Levovist are easily collapsed under the high MI condition, so it is hard to visualize the bubbles successively. Dynamic Flow has solved the above problems. Dynamic Flow uses Doppler technique, but Dynamic Flow image has higher resolution and less blooming area than the conventional Doppler image. Dynamic Flow doesn't display THI, so the detectability of bubbles is superior to the other mode as far as Levovist. Dynamic Flow is so sensitive that it can show the bubbles in real time. Advanced Dynamic Flow(ADF) has greatly improved in sensitivity and presentation ability from conventional Dynamic Flow. In the contrast application, ADF image looks very similar to B-mode image thanks to the high resolution and wide dynamic range, in spite of using Doppler technique. Detectability of the bubbles is much better than conventional Dynamic Flow. ADF can detect bubbles in much smaller vessels and has much more penetration, with uniform enhancement from shallow to deep regions. ADF is very suitable for Levovist. ADF can also be used without contrast agents. ADF can show small tiny vessels with high resolution and high frame rate. Directional information can be added by different colors. The sensitivity is close to conventional Color/Power Doppler images. PMID- 12766301 TI - [Therapeutic technology utilizing focused ultrasound] AB - Ultrasound has two major biological effects potentially to be utilized for tumor treatment: heating and cavitational effects. In coagulation treatment, the tissue in the focal spot is heated above the coagulation temperature in a relatively short period of time with high intensity focused ultrasound so that the irreversible change in tissue is completed much earlier than the temperature distribution blurs due to heat conduction. However, the smallness of the coagulation volume formed by an ultrasonic shot causes the problem of low treatment throughput. The split-focus technology, in which the focal spot is enlarged in the lateral rather than longitudinal direction, multiplies the coagulation volume so as to solve this problem. Sonodynamic treatment was proposed based on the recent in vitro and in vivo experimental findings that ultrasonic cavitation can activate certain porphyrins and thereby induce significant antitumor effects. It was also found that the ultrasonic intensity threshold for producing cavitation can be significantly reduced by superimposing the second harmonic onto the fundamental. The threshold to form a focal lesion in murine liver tissue was reduced by orders of magnitude, especially in combination with administration of a certain xanthene dye. PMID- 12766302 TI - Emission CT Image Reconstruction Using the Augmented Lagrangian Method with a Modified Form of Nonnegativity Constraints. AB - The augmented Lagrangian method was applied to emission CT image reconstruction to attain the satisfactory elimination of negative pixels using a modified form of the original nonnegativity constraints. A penalty function was included in the augmented Lagrangian method to suppress image noise due to statistical noise in the projection data. A slight change in the original constraint form achieved almost complete nonnegativity without loss in the closeness of reconstruction to the original image. PMID- 12766303 TI - Monte Carlo simulation for PET scanners and shields. AB - A Monte Carlo simulation code was developed for simulating PET scanners with the Monte Carlo program package GEANT. The present simulation code can handle not only conventional types of PET scanners, but also any complex detector systems with arbitrary geometrical configuration. All the relevant interactions of photons and electrons are taken into account in all the defined objects while optical tracking in the scintillation crystals is approximated by simple analytical simulation. In addition to basic PET scanner performance factors, such as sensitivity and scatter fraction, valuable but un-measurable information, such as photon trajectories and interaction position distribution, can be obtained and represented graphically in various ways. This simulation code has proved useful in analyzing the physics characteristics of existing commercial PET scanners and related shields, and in design studies of new PET scanners. PMID- 12766304 TI - [Physical and imaging properties of heavy metal filter combinations in chest radiography] AB - We have investigated the performance characteristic of the heavy metal filters with higher atomic numbers by comparing their patient exposures, tube loadings, radiographic contrasts, and noise Wiener spectra with those of a combination of copper and aluminum filter which has been used widely in chest radiography. Seven heavy metal filters were used for this study. As for a tungsten filter, two filters different in thickness were used. One is 0.05 mm thick, and the other 0.10 mm. The other metal filters were respectively combined with a tungsten filter with a thickness of 0.05 mm. Among the all filters investigated, tube loading of tungsten filter with 0.05 mm thick is minimum. Tungsten with 0.1 mm thick and tungsten with 0.05 mm+ barium show the larger advantages in patient exposure than those of the other filters. It was found that the magnitude of patient exposure varied slightly with the difference of image receptor used. The use of heavy metal filters showed the small advantages in the patient exposure reduction ( approximately 20%) compared with that of conventional copper filter. The use of heavy metal filters showed increasing tube loading ( approximately 5 times) compared with that of conventional copper filter. Noise Wiener spectra of heavy metal filters showed comparable with those of conventional copper filter. In conclusion, the use of heavy metal filter combinations offer no significant advantages over optimal conventional filters. PMID- 12766305 TI - [Comments on the primary and scatter dose-spread kernels used for convolution methods] AB - The basic primary and scatter dose-spread kernels used for convolution methods are usually produced by Monte Carlo simulations with the interaction point forced to the center of a large water phantom. This paper proposes another type of basic kernel, with which perfectly accurate primary and scatter absorbed dose calculations can be performed within a semi-infinite water phantom under certain irradiation conditions. PMID- 12766306 TI - [Observation of Cherenkov Radiation from Aquarium Irradiated with Co-60 Gamma rays] AB - Cherenkov radiation in water at a nuclear power plant is caused by a nuclear fuel rod and is well known generally. If students can observe Cherenkov radiation at school easily, they can be impressed by the fascinating radiation. Moreover the observation may bring about interest in radiological physics profoundly. A few years ago, management of the Co-60 gamma-ray irradiation apparatus was transferred to Nagoya university school of health sciences from the related hospital. Therefore we have examined the system to observe the Cherenkov radiation in water from secondary electrons generated by Co-60 gamma-rays. At first, the Cherenkov radiation in the aquarium was led to the corridor outside the irradiation room using a mirror, and observed directly while avoiding exposure. Secondly photographs of the Cherenkov radiation from various angles were taken under conditions which consisted of several irradiation fields and pass lengths of gamma-rays in water, and were compared with each other. Our method for observing the Cherenkov radiation may be useful enough for students to raise their dedication in radiological physics study. PMID- 12766308 TI - [CyberKnife] AB - The CyberKnife is an image-guided robotic system designed for stereotactic radiosurgery. This system uses a lightweight, x-band linear accelerator, computer controlled robotic arm, a pair of orthogonal x-ray imagers (TLS: Target Locating System), and a computer workstation. During the treatment, the TLS determines the location of the lesion and communicates these coordinates to the robot. The robot adjusts the position of the beam to the target. The accuracy of this system is 0.7 mm (median) at Osaka University. The CyberKnife system offers new options for radiosurgery/therapy. Stereotactic fractionated radiotherapy can now be performed with the same accuracy as single-fraction stereotactic radiosurgery. The frameless nature of CyberKnife allows tumors in the chest and abdomen to be treated as well. The real time tracking system option enables one to treat tumors that move with respiration, such as lesions in lung. Tumors in the lower spine, pancreas, and lung have already been treated in the USA. A description of the components, accuracy, and future of the CyberKnife will be presented. PMID- 12766307 TI - [Intensity modulated radiation therapy] AB - A preliminary goal of radiation therapy is to deliver radiation doses that conform to targets, while minimizing dose to surrounding normal tissue. Intensity modulated radiation therapy (IMRT) delivered with a multileaf collimator (MLC) is a promising resource to achieve conformal therapy. A medical linear accelerator (linac) equipped with an IMRT system can increase dose to the target and decrease dose to normal tissue. These capabilities may be significantly enhanced with dose optimization techniques. IMRT integrates rapidly evolving imaging, computer, optimization, and treatment delivery technologies. Resulting systems fundamentally distinguish themselves from prior technology. Thus, new quality assurance (QA) procedures are needed. PMID- 12766309 TI - [Gated Radiotherapy] AB - Recent external radiotherapy requires precise localization of the target because advance in diagnostic imaging has made it possible to visualize a tiny tumor which would be curable with focused high dose irradiation. However, tumors in respiratory and bowel organs have been difficult to be given the high dose because of 1 to 3 cm movement during delivery of irradiation. Respiratory-gating techniques have been used with medical linear accelerators and particle therapy machines. Real-time tumor-tracking radiotherapy has been realized using fluoroscopic x-rays, internal gold-markers, and pattern recognition technology. Advantage and disadvantage of each gating technique have been realized. Active breath control method would be a cost-effective way of precise treatment without gating. More work is required to find the relationship between abdominal wall and internal movement of the tumor in many respiratory-gating radiotherapy and between the internal markers and target volume in real-time tracking radiotherapy. PMID- 12766310 TI - [Three-dimensional conformal radiotherapy for extracranial tumors using a stereotactic body frame] AB - This paper was reviewed to evaluate the feasibility of three-dimensional (3-D) conformal radiotherapy for extracranial tumors, especially for solitary lung tumors using a stereotactic body frame. To extend the technique of stereotactic irradiation for intracranial tumors, accurate body fixation and regulation of internal target motion are essential. In our study, a stereotactic body frame was used, and daily setup accuracy was verified. As a result, its setup accuracy was maintained within 0-8.5 mm (Ave=2.5mm). In our initial clinical experiences for thirty-two patients with 6-10 non-coplanar static beams, forty or 48 Gy was irradiated. During the follow-up of 4-27 (Average=11) months, twenty-nine (94%) tumors were locally controlled without any symptomatic complications. Recently, respiratory-gated irradiation systems, CT-linac systems, a real-time tumor tracking system, Cyber-knife, and C-arm linac were developed. With all these techniques, stereotactic irradiation for extracranial tumors are future direction of three-dimensional conformal radiotherapy. PMID- 12766312 TI - [A monitor unit verification calculation independent on RTP systems and output factor] AB - In order to confirm quality assurance and quality control in radiation therapy, it is common to perform a monitor unit verification calculation independent on RTP system. Generally, this verification calculation is often to perform at manual calculation level separately from RTP system. Characteristics of output factors included in this calculation were explained on the basis of the situation in the world. Khan et al. had proposed the concept of collimator scatter factor and phantom scatter factor. The validity of these factors was addressed in of role played in dose monitor unit calculation. If we will utilize SMR, which was derived from zero-area phantom scatter factor, and zero-area TMR in dosimetry, a simple and accurate Clarkson integration is possible. These scatter factors and Clarkson integration is able to use not only for monitor unit calculation in a simple treatment technique, but also in an excellent technique as IMRT. These factors had been also used in the convolution method to improve an accuracy of dose calculation. These factors are not family in Japan, but it is expected to be got an agreement of the usage in future. PMID- 12766313 TI - [Annual change in calibration constant of ionization chamber used in diagnostic radiology] AB - Committee on Radiological Protection of Japan Radiological Society (JRS) is making a database on patient doses for typical radiographies for introducing the guidance levels in Japan. The Committee reported necessity of a protocol on dosimetry in diagnostic radiology for introducing the guidance levels. Although a protocol on dosimetry in radiation therapy is established, there is no protocol on dosimetry in diagnostic radiology in Japan. Then, as a first step to establish a protocol on dosimetry in diagnostic radiology, annual changes in a calibration constant of an ionization chamber as a practical standard dosimeter has been investigated for 4 years. Changes in calibration constants for 4 years are within 2% for X-rays with effective energies than 32 keV. From these results, practical frequencies of calibrations for practical standard dosimeters are discussed for maintain of their precision. If an accuracy of practical standard dosimeters is 5% for diagnostic radiology, frequencies of their calibrations may be once in a year or two years. PMID- 12766311 TI - [Three-dimensional conformal radiotherapy with multi-leaf collimator] AB - In radiation therapy, it is very important to concentrate the high dose to the clinical target volume. For this purpose, S. Takahashi developed the dynamic conformal radiotherapy in 1960. In the first half of 1990' this conformal technique with multi-leaf collimator (MLC) was gradually becoming popular, because the three dimensional extent of the clinical target volume and the surrounding normal tissues was accurately recognized by the use of the serial CT images, and the 3D-treatment planning was considerably easily obtained by the development of computer-technique. The 3D-CRT (conformal radiation therapy) based on the 3D-treatment planning in a broad sense means the coplanar or non-coplanar irradiation with multi-leaf collimator, and at present it is one of the most reasonable treatment techniques. In order to correspond the high dose region to the clinical target volume as possible, IM (intensity modulation) technique was introduced to the 3D-CRT using MLC in the second half of 1990'. Therefore, the static CRT is routinely used not only in Europe/ USA but also in Japan. But, it is often very difficult to obtain the concave high dose region in static CRT with MLC, even though the IM-technique is applied. At this occasion, the concave high dose region is easily obtained by the use of dynamic CRT such as the hollowed out technique developed by S. Takahashi in Japan. The dynamic CRT technique is also useful with the static IM-CRT technique. There are two main aims of conformal radiotherapy: First, the protection of the surrounding normal tissues, and second, improvement of the local control rate. Usually the second aim can't be easily achieved, because the tumor control probability curves are often flat. The DVH concept is usually valuable to choose the optimal treatment planning. But, it is difficult to find the optimal dose-distribution, when two DVH-curves cross each other on the middle range of irradiated dose. On that occasion, the NTCP-concept is useful. From the standpoint of the tolerable dose, normal organs are classified into two types: parallel and serial. Therefore, two types of the NTCP-formula should be prepared to apply the NTCP-concept clinically. PMID- 12766314 TI - Obsessive-compulsive disorder in childhood and adolescence. AB - Obsessive-compulsive disorder (OCD) is a common psychiatric condition during childhood and adolescence, which continues to be underestimated and undertreated. For these reasons, it constitutes a primary cause of major disabilities in those ages and, sometimes, of permanent impairments later on. In these last few years, childhood and adolescence OCD has attracted an increasing focus which has promoted a deeper awareness of this illness, a better recognition with earlier interventions, as well as the set-up of more tailored and specific strategies, including psychotropic drugs. The aim of this paper is to present a critical review of paediatric OCD, with a special attention towards the most compelling reports available up to now and towards the most interesting areas for future research. PMID- 12766315 TI - Gender differences in dissociation. A dimensional approach. AB - Considering that epidemiological research on dissociative disorders has suggested a 9 to 1 predominance of female cases, this study investigated the relationship between gender and dissociation using a dimensional approach. A total of 2,153 participants from different diagnostic groups completed the Dissociative Experience Scale. In order to control for the confounding effect of current psychopathology a subgroup 790 subjects additionally completed the SCL-90. We did not find any differences in the general or pathological dissociation scores. Hypothetical gender differences in dissociative psychopathology were not a function of diagnostic categories. There were no significant sex differences in the distribution of high dissociators. Our findings suggest that men and women do not generally differ in dissociative psychopathology. The implications for future investigations on the epidemiology, etiology, and psychobiology of dissociative symptoms are discussed. PMID- 12766316 TI - Relationship and differential validity of alexithymia and depression: a comparison of the Toronto Alexithymia and Self-Rating Depression Scales. AB - Theoretically, the constructs of alexithymia and depression share many common characteristics. Empirically moderate correlations between measures of alexithymia and depression have been found, hence it has been argued that the Toronto Alexithymia Scale (TAS) may be, at least in some part, just another measure for depression. The purpose of this study was to evaluate the construct validity of alexithymia and to analyze the relationship between alexithymia and depression. Exploratory and confirmatory factor analyses were conducted on the combined items of the TAS-20 and the Self-Rating Depression Scale (SDS) in a psychosomatic and alcoholic inpatient sample (n = 199) and a sample of normal adults (controls, n = 174). The exploratory factor analysis in the patient sample yielded a 4-factor structure. Within each factor there was no overlap between the items of the TAS-20 and the SDS. Two factors were comprised of items of the TAS 20 and two factors consisted of items of the SDS. This 4-factor model also showed an acceptable fit for the data of the normal sample in a confirmatory factor analysis. Moderate correlations between the TAS- 20 and SDS total scores and some factors were found. Both instruments, the TAS-20 and the SDS, seem to measure distinct constructs and are not just different measures for the same underlying construct. This provides support for the differential validity of the alexithymia and depression constructs and is in accordance with previous findings. PMID- 12766317 TI - Personality disorders and self-perceived quality of life in an elderly psychiatric outpatient population. AB - The aim of this work was to assess the impact of personality disorder and traits on the subjective assessment of quality of life by a sample of elderly psychiatric outpatients (60 years or more). Sixty of the total number of subjects interviewed were included in the test sample since they met the Vragenlijst Klinishe Persoonlijkheid (VKP) criteria for personality disorder. A control sample was also formed, composed of 50 subjects without personality disorders or traits. Quality of life was assessed using the LEIPAD questionnaire. Personality disorder diagnoses were prevalently distributed in cluster C (25%) and in cluster A (20%). A total of 48.3% of patients (n = 29) had more than 1 personality disorder. Sixty percent of the sample (n = 36) presented with comorbid Axis I psychiatric disorders, in most cases depressive pathologies. Comparing the case and control groups, worse LEIPAD scores were achieved by subjects with personality disorders. Between clusters of personality disorders, the worst scores were observed in subjects with more than 1 personality disorder diagnosis. Elderly people who presented with a personality disorder were more vulnerable to late-life distress, especially those with more than one diagnosis. In keeping with linear regression analysis, widowhood, divorcehood and living with others appear to be variables able to negatively influence quality of life in this population. While the VKP needs to be adapted to the application in the elderly, it does seem to be a particularly useful screening instrument. PMID- 12766318 TI - Combining the Rorschach test and the Temperament Character Inventory: a new perspective on personality assessment. AB - BACKGROUND: The numerous reports on research involving the clinical assessment of personality in axis I disorders highlight the importance of temperament features in the current approach to all mental disorders. However, the available instruments of personality assessment have many limits. Self-administered questionnaires depend on the patient's insight, and projective instruments (i.e. the Rorschach test) often lack objectivity. This study compared the results of personality assessment with the Temperament and Character Inventory (TCI) and the Rorschach test to verify their validity. PATIENTS AND METHODS: TCI and Rorschach tests were administered to a wide sample of patients (n = 180) in a short period. The most common Rorschach siglatures were correlated with the TCI raw scores using the Pearson correlation test. RESULTS: All TCI temperament dimensions and facets displayed at least two correlations with Rorschach siglatures. The description of each dimension and facet of the TCI obtained with the interpretation of Rorschach siglatures was consistent with its original meaning. CONCLUSIONS: The TCI and Rorschach tests adequately validated each other. In the future, the administration and integration of these tests will overcome the biases of both. Further, the theoretical bases of the TCI could facilitate the study of psychological functions, whereas the psychodynamic bases of the Rorschach test provide an in-depth insight into temperament traits. PMID- 12766319 TI - A study of the phenomenology of panic attacks in patients from India. AB - We studied the symptom structure of panic attacks in 94 patients attending an outpatient psychiatric clinic. Autonomic symptoms of anxiety were reported by the majority of the panic patients. As compared with studies from the West, certain cognitive symptoms, such as fear of loss of control and depersonalization, and vestibular symptoms were endorsed by fewer panic patients. The majority of our panic patients tended to seek help initially from physicians and medical specialists. The majority of the patients also did not experience some DSM-IV symptoms as part of their panic attacks. Comorbid psychiatric conditions influenced the symptom structure of panic attacks. A higher number of agoraphobics as compared with patients with uncomplicated panic disorders had the symptoms of fear of loss of control and fainting. Our findings support the notion that the experience of panic attacks is variable and differs across cultures. PMID- 12766320 TI - Amplification and attribution styles in somatoform and depressive disorders--a study from Bangalore, India. AB - OBJECTIVE: The present investigation aimed to study attribution styles and somatosensory amplification among patients suffering from somatoform and depressive disorders. METHODS: Two groups of 30 patients with diagnoses of somatoform disorder and depressive disorder, respectively (ICD-10 DCR), and one group of 30 normal controls were recruited. The study patients were assessed using the symptom interpretation questionnaire, somatosensory amplification scale, and scales for assessing alexithymia and illness attitudes. RESULTS: The somatoform and depressive disorder patients had greater recent symptom experience than the normal group. The somatoform disorder group had higher somatic attribution scores, the depressive disorder sample had higher psychological attribution scores, and the normal group had higher normalizing attribution scores than the two other groups. Somatoform disorder patients had higher mean amplification scores than depressed patients, who in turn had higher scores than normals. Correlation analyses showed somatic attribution and certain illness attitudes to be closely associated in all three groups. Recent symptom experience was associated with amplification in the somatoform disorder group alone. Recent symptom experience, a diagnosis of somatoform disorder and lower normalizing attribution scores predicted amplification. DISCUSSION: These findings indicate that somatoform and depressive disorder patients and normals differ from each other in their attribution styles. There is a clustering of attributes among somatoform disorder patients that include greater symptom experience, which is somatically attributed, and is associated with excessive illness worry, concern and preoccupation with bodily symptoms, and a fear of having or developing a disease. On the other hand, depressed patients and normal subjects who do have a somatic attribution style (though, as a group, they have lower somatic attribution scores than the somatoform disorder group), also harbor hypochondriacal beliefs and related attitudes. PMID- 12766323 TI - Gender differences in methamphetamine-induced mRNA associated with neurodegeneration in the mouse nigrostriatal dopaminergic system. AB - In this report female and male CD-1 mice were treated with a neurotoxic regimen of methamphetamine (MA) to compare gender differences in striatal dopamine depletion and concordant changes in mRNA markers of the transforming growth factor-beta injury response associated with neurodegeneration. Striatal dopamine concentrations of MA-treated female mice were less depleted and significantly greater than that of identically treated males. Associated with this gender difference in striatal dopamine depletion were significantly decreased mRNA levels of plasminogen activator inhibitor-1 and a trend for increased (p = 0.06) mRNA levels of glial fibrillary acidic protein within females. No statistically significant differences between MA-treated female and male mice were obtained in mRNA levels for transforming growth factor-beta, transforming growth factor-beta type 2 receptor, activin-like kinase-5 or fibronectin. These data demonstrate the presence of changes in two specific molecular markers of the transforming growth factor-beta injury response which are in accordance with gender differences in MA induced striatal dopamine depletion. The results suggest that the neuroprotective advantage displayed by females may in part be related to reductions in the transforming growth factor-beta injury response as indicated by decreased mRNA plasminogen activator inhibitor-1 and an increased response of reactive astrocytes which promote neuronal survival as indicated by augmented glial fibrillary acidic protein mRNA levels. PMID- 12766321 TI - Musical hallucinations in schizophrenia. 2. Relations with verbal hallucinations. AB - The musical hallucinations reported by 33 schizophrenics who fulfilled ICD-10 diagnostic criteria were assessed semiologically. The subjects were 24 men and 9 women. The duration of 48 episodes of musical hallucinations varied. They occurred at all times during the course of schizophrenia, and 9 of them (18.8%) occurred before the diagnosis was made. All of the episodes could be divided into three stages: 17 (35.4%) were first stage, close to obsession, 5 (10.4%) were second stage, close to Schneiderian first-rank symptoms, and 26 (54.2%) were third stage, which is the autochthonous experience. The sounds were in a subjective space in 38 episodes and in the majority of cases, their content was familiar. The second stage was associated with xenopathic experience ("gemachtes Erlebnis") and audition of thought, and words were added to melodies that normally had no lyrics. Musical hallucinations in schizophrenia are pseudohallucinations that originate in memory representations, and they may undergo a transition to true hallucinations. The authors think that the three stages are related to the severity of the disease. PMID- 12766322 TI - Repeated estradiol treatment prevents MPTP-induced dopamine depletion in male mice. AB - Epidemiological data suggest that the steroid hormone 17beta-estradiol plays an important role in protecting the brain from neurodegenerative processes, including that causing the loss of dopamine (DA) neurons in Parkinson's disease. Determining the mechanisms of neuroprotection in experimental systems may facilitate the development of estrogenic therapies for these diseases. The present study sought to further investigate the mechanism of the neuroprotective effect of 17beta-estradiol in a murine model of Parkinson's disease, i.e. 1 methyl- 4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced striatal DA depletion. Consistent with previous findings, 17beta-estradiol was found to inhibit MPTP induced DA depletion under a dosing regimen (repeated daily administration) that mimicked physiological levels of the steroid. However, high doses of the steroid administered repeatedly or acutely failed to inhibit toxicity, as did 17alpha estradiol. These data suggest that the neuroprotective effect of 17beta-estradiol was mediated through an interaction with one of the nuclear estrogen receptors, and is not the result of an antioxidant action. In order to realize the therapeutic potential of the neuroprotective effect of 17beta-estradiol for Parkinson's disease, it will be necessary to identify synthetic estrogen receptor modulators that lack the activity of the steroid on peripheral tissue. In this study, raloxifene failed to mimic the neuroprotective effect of 17beta-estradiol against MPTP toxicity. Thus, exploration of new compounds with different pharmacological and/or physiochemical properties is warranted. PMID- 12766325 TI - Regulation of gonadotropin releasing hormone release by neuropeptide Y at the median eminence during the preovulatory period in ewes. AB - The median eminence (ME) of the hypothalamus is known to be an important brain site where hypophysiotropic release might be regulated by excitatory and inhibitory signals impinging on their neuronal terminals. Since a role for neuropeptide Y (NPY) on preovulatory luteinizing hormone (LH) release has been suggested, we hypothesized that NPY might act at the ME to control preovulatory gonadotropin-releasing hormone (GnRH) release and thus the onset of the preovulatory surge of LH. To examine this possibility, we used the ewe as an animal model to determine: (a) immunocytochemical distribution of GnRH and NPY in the ewe ME; (b) changes in in vivo release of NPY and GnRH using ME push-pull cannula (PPC) perfusate samples, as well as in plasma LH, during the luteal, follicular and preovulatory phases of a synchronized estrous cycle, and (c) effects of ME perfusion of NPY or a Y1-NPY antagonist, or an NPY antiserum on in vivo release of ME-GnRH and plasma LH during a synchronized follicular phase. Immunolocalization reveals a dense plexus of beaded GnRH-containing neurites in the arcuate nucleus and in its vicinity, the pituitary stalk and the palisade. In contrast, a dense plexus of NPY-containing neurites occurs in the internal layer, with occasional fibers found in the intermediate and lateral external zone of the ME. In the area between the lateral internal and lateral external layers, both NPY and GnRH-containing processes were found, thus providing opportunities for synaptic and/or paracrine interactions between NPY- and GnRH-containing neurons. Hormonal analysis indicated that a synchronized preovulatory surge of LH is elicited within a 2-hour window by the sequential implantation and removal of silastic-encased estradiol (E2) or progesterone (P4) implants. In this paradigm, there was a parallel increase in ME release of both NPY and GnRH preceding the synchronized LH surge. The onset of this synchronized LH surge was advanced by ME perfusion of exogenous NPY and was both delayed and blunted by ME perfusion with the NPY antagonist (both were perfused through the PPC probe for 2 h, starting 2 3 h before the expected onset of the LH surge). In addition, NPY perfusion in the ME increases, while perfusion of the Y1-NPY antagonist or of the NPY antiserum decreases ME-PPC GnRH content and plasma levels of LH in early follicular ewes. Finally, perfusion of NPY antiserum during an ongoing LH surge disrupted LH release. These results suggest that interactions between NPY and GnRH neurons are important in controlling the timing, magnitude and maintenance of the preovulatory LH surge. PMID- 12766324 TI - The effect of estradiol in the striatum is blocked by ICI 182,780 but not tamoxifen: pharmacological and behavioral evidence. AB - Estradiol in the striatum enhances amphetamine (AMPH)- or KCl-stimulated dopamine (DA) release and the pacing of sexual behavior in the female rat. These effects of estradiol in the striatum are rapid, steroid specific and thought to be mediated by a G protein-coupled membrane receptor for estradiol. In the current experiments, we examined whether two antiestrogens, ICI 182,780 (ICI) and tamoxifen (TAM), affect the enhancement by estradiol of (1) AMPH-induced DA release from striatal tissue in vitro, and (2) paced mating behavior in the female rat. The steroidal antiestrogen ICI significantly blocked the effect of estradiol on AMPH-induced striatal DA release. In contrast, the nonsteroidal, triphenylethylene antiestrogen TAM did not block the effect of estradiol, even when a concentration 10 times the concentration of estradiol was delivered to the tissue. Neither of the compounds showed estrogenic action when tested in the absence of estradiol. When implanted into the dorsolateral striatum, ICI inhibited the effect of estradiol on pacing of sexual behavior, while TAM did not. The specific effects of ICI, but not TAM, on striatal DA release and pacing behavior provide further information about the binding site for estradiol in the striatum. PMID- 12766326 TI - Estradiol and testosterone have opposite effects on microtubule polymerization. AB - We have reported earlier the purification of tubulin from a plasmalemmal microsomal fraction derived from rat hippocampus using an estradiol (E(2)) affinity column and the specific binding of tubulin to both E(2) and testosterone (T). To further investigate the effect of E(2) and T on the function of this protein, changes in microtubule polymerization as a result of exposure to the steroids were examined in this study, using both pure tubulin and rat hippocampal primary cell cultures. First, pure tubulin was incubated with or without steroids for 30 min on ice followed by polymerization at 37 degrees C. The numbers of microtubules formed were counted from electron microscopic pictures. The results showed that at 30 min of polymerization, 10 nM, 30 nM and 30 microM of E(2) inhibited microtubule assembly by -70%, -94%, and -92%, respectively (p < 0.01), while T at the same three concentrations stimulated it by +83%, +66%, and +121%, respectively (p < 0.05). The inhibitory effect of E(2) and the stimulatory effect of T were observed at 15, 30 and 60 min of the polymerization process. Next, primary cell cultures from 17-day rat fetus hippocampal tissues were treated with the steroids and polymerized microtubules (Triton X-100 resistant) were examined by immunocytochemistry. The results demonstrated that 60 min of E(2) treatment (10 nM) decreased the intensity of the immunolabeling of polymerized microtubules. The effect of T at nM concentration was not significant though it increased the immunolabeling at microM concentration. Of great significance was a remarkable inhibition by T of the well-established depolymerization effect of colchicine in both the pure tubulin assay and the cell culture model, while E(2) was not effective. In an effort to pursue the possible mechanism(s) of the effect of E(2) and T on microtubule formation, we found that T only inhibited the microtubule depolymerization process without affecting the rate of polymerization. In contrast, E(2) modifies only the polymerization process without altering the depolymerization. Overall, these data indicate that E(2) and T may be considered as novel regulators of microtubule dynamics and thereby controlling cytoskeleton function in cells. PMID- 12766327 TI - Thyrotropin-releasing hormone as a mediator of the central autonomic pathway controlling ovarian function. AB - We studied the effect of thyrotropin-releasing hormone (TRH) applied centrally on the sympathetic activity of the ovary in female rats. Intracerebroventricular (i.c.v.) administration of a dose of 25 ng/kg weight produced an increase in noradrenaline (NA) content at the ovary after 5 days of hormone administration. However, higher doses in a range up to 500 ng/kg weight decreased NA content at the ovary. At the celiac ganglia (where the cell bodies of sympathetic neurons projecting to the ovary originate) there was an accumulation of NA in spite of a decrease in tyrosine hydroxylase activity (T-OH). After cold exposure, opposite effects on T-OH activity and no effects on NA in ganglia and in ovary were obtained. Besides, i.v. injection of TRH only induced a decrease in ovarian NA. In contrast to the increase in T(3) plasma levels obtained after the cold-stress procedure, none of the i.c.v. doses of TRH used produced changes in T(3) plasma levels, strongly suggesting that the effect on sympathetic activity is mediated by a central effect of TRH acting as a putative activator of ovarian sympathetic nerves. PMID- 12766328 TI - Cell differentiation of gonadotropin-releasing hormone neurons and alternative RNA splicing of the gonadotropin-releasing hormone transcript. AB - Two different, yet related issues regarding gonadotropin-releasing hormone (GnRH), i.e. the development and differentiation of hypothalamic GnRH neurons and the alternative RNA splicing of GnRH gene transcripts, are addressed in the present review. Using the immortalized GnRH-producing GT1 cell line, we found that activation of protein kinase C (PKC) with 12-O-tetradecanoylphorbol-13 acetate induces morphological and functional differentiation of these neurons. Specific isoforms of PKC are involved in neurite growth, cell migration and synaptic contacts and involve different signaling pathways. Using an in vitro splicing assay with HeLa nuclear extract, we found that excision of the first intron of the GnRH primary transcript is attenuated in non-GnRH-producing cells, but not in GnRH-producing cells such as GT1. This attenuation was relieved by exonic splicing enhancers located in the GnRH exons 3 and 4. Interestingly, addition of nuclear extract derived from GT1 cells further increased the excision rate of intron A, indicating that GnRH neurons contain TRANS-acting splicing factors. Extensive biochemical analysis indicates that Tra2alpha, a serine/arginine-rich RNA-binding protein, and other cofactors are likely involved in mediating neuron-specific excision of intron A from the GnRH primary transcript. An understanding of the GnRH neuron-specific splicing machinery provides critical insight into the molecular mechanism of GnRH gene regulation and consequently of mammalian reproductive development. PMID- 12766329 TI - Point surveillance of Staphylococcus aureus carriage among medical staff in Infectious Diseases Hospital, Kuwait. AB - OBJECTIVE: To investigate the carriage of Staphylococcus aureus among doctors and nurses in Infectious Diseases Hospital, Kuwait, following the detection of 3 cases of methicillin-resistant S. aureus (MRSA). MATERIALS AND METHODS: A total of 260 nasal and throat swabs were obtained from 19 doctors and 111 nurses and cultured for the carriage of S. aureus. Forty-three S. aureus were identified based on their growth characteristics on mannitol-salt agar, catalase and tube coagulase and DNA hydrolysis. The isolates were tested for susceptibility to antibacterial agents and typed by phage typing; plasmid analysis and pulsed-field electrophoresis were carried out to determine their relatedness. RESULTS: Of the 19 doctors, 4 (21%) had nasal carriage while only 1 of them had a throat carriage. Sixteen (14.4%) nurses carried S. aureus in their noses and 20 (18%) in their throats. The combined nasal carriage rate for both doctors and nurses was 15.8%, and combined throat carriage was 16.6%. None of them carried MRSA. The isolates were resistant to penicillin G (90%), tetracycline (23.3%), erythromycin (9.3%) and cadmium (100%). Typing of the isolates showed a variety of phage types, plasmid and pulsed-field gel electrophoresis patterns. DISCUSSION: None of the doctors or nurses carried MRSA. Typing of the methicillin-susceptible strains that they carried demonstrated that the S. aureus were different, indicating an absence of a dominant clone capable of spreading. It is important to maintain a low carriage of S. aureus among health-care workers. PMID- 12766331 TI - The protein polymorphism of haptoglobin in Korean elite athletes. AB - OBJECTIVE: To investigate protein polymorphism of the haptoglobin (Hp) and the relationship between Hp phenotypes and anthropometric or biochemical parameters in elite Korean male athletes. MATERIALS AND METHODS: Serum samples were collected from 120 Korean male elite athletes. The Hp phenotypes were determined by polyacrylamide gel electrophoresis, followed by peroxidase staining. Then anthropometric or biochemical measurements were made: body composition, blood pressures, ventilatory responses, cholesterol (total, LDL cholesterol and HDL cholesterol), triglyceride, apolipoprotein A1, lipoprotein (a), creatine phosphokinase and lactate dehydrogenase. RESULTS: The gene frequencies of the Hp1 1, Hp2-1 and Hp2-2 phenotypes in Korean male athletes were 12, 37 and 51%, respectively; this polymorphism was significantly associated with the VO(2max) index in the athletes. An excess of the Hp1 allele was also observed in marathon runners compared with the other sporting activities, although it did not have any statistical significance. CONCLUSION: Hp polymorphism exists in elite Korean male athletes and Hp phenotype may be a useful marker for endurance performance in these male athletes. PMID- 12766330 TI - Sectorial technetium-99m-dimercaptosuccinic acid scintigraphy for monitoring the effect of extracorporeal piezoelectric lithotripsy for calyceal calculi on regional renal function. AB - OBJECTIVE: To apply a semiquantitative method for analysis of technetium-99m dimercaptosuccinic acid ((99m)Tc-DMSA) renal scintigraphy for monitoring the effect of extracorporeal piezoelectric lithotripsy (EPL) in patients with calyceal stones on regional kidney function and to check whether EPL had caused any deleterious effect on the target calyceal renal parenchymal function. PATIENTS AND METHODS: Forty patients (mean age 35 years) suffering from calyceal stones documented by abdominal plain radiography, intravenous urogram or abdominal ultrasound were studied. All patients were treated by EPL. (99m)Tc-DMSA scan was performed before and 4 weeks after EPL. Sector analysis involved calculation of the relative function of the target calyx to the function of the ipsilateral kidney and the relative function of the treated kidney to global renal function. RESULTS: The stone sizes were 6-11 mm in diameter and 11 were located in the upper, 13 in the middle and 16 in the lower calyx. After EPL, the overall stone clearance rate was 85% (100% for calculi in the upper and middle calyces, 62% for lower calyces). The sector analysis did not show statistically significant change of the relative regional (calyceal) or whole kidney function between the pre- and post-EPL (99m)Tc-DMSA scans. Using sector analysis, EPL appeared to be a safe modality and its usage was not associated with any untoward effect on calyceal or whole kidney function. CONCLUSION: Sector analysis of (99m)Tc-DMSA renal scan is a simple semiquantitative method for monitoring regional changes of kidney function after EPL for treatment of calyceal stone. PMID- 12766332 TI - Modified Giemsa staining for rapid diagnosis of malaria infection. AB - OBJECTIVES: To develop and evaluate a rapid method for the diagnosis of malaria infection by microscopy of stained blood films. SUBJECTS AND METHODS: Blood specimens were collected from randomly selected confirmed malaria cases (n = 75) and suspected malaria cases (n = 175). The microscopy was done on each set of blood films stained by modified and the standard Giemsa staining methods. RESULTS: All the 75 previously diagnosed malaria cases were confirmed by the microscopy of blood films stained by both methods. Forty-nine (28%) of the 175 cases suspected for malaria infection showed malarial parasites on microscopy of blood films stained by both methods. However, due to homogeneous staining and clearer background of the blood films it was possible to determine the parasite species in 65% of the cases on microscopy of the thick films stained with the modified method compared to only 20% with the standard method. Further, the turnaround time for reporting the microscopy test result was 15-20 and 45-50 min with modified and standard staining methods, respectively. CONCLUSION: Our data showed that performance of the modified staining method in detecting malarial parasites was comparable to that of the standard staining method. Moreover, the modified staining method was rapid, easy to use, and reliable. PMID- 12766333 TI - Comparison of lactose intolerance in healthy Kuwaiti and Asian volunteers. AB - OBJECTIVE: To study and compare the incidence of lactose intolerance among Kuwaiti and Asian healthy volunteers as measured by breath hydrogen level following challenge with lactose drink. SUBJECTS AND METHODS: The study involved 70 Kuwaiti and 79 Asian healthy volunteers. The volunteers were physicians, medical students and other hospital workers. The study was carried out prospectively at Amiri Hospital, Kuwait. None of the volunteers was sick or had taken antibiotics or any other drug that could influence breath hydrogen level 2 weeks prior to the study. After an overnight fast 20-ml baseline samples of exhaled air were collected from each volunteer prior to oral administration of 40 g of lactose drink. Following this, the same amount of exhaled air was collected at 30-min intervals for 2 h. All samples were analyzed for hydrogen level using a Quintron microlyzer. Hydrogen level of 20 ppm more than baseline value was considered positive. RESULTS: The basal breath hydrogen levels were 12.49 +/- 8.4 and 6.97 +/- 6.9 for Kuwaitis and Asians, respectively. Thirty-three (47%) of the 70 Kuwaitis and 46 (58%) of the 79 Asians were positive for the breath hydrogen test. The clinical symptoms of flatulence, abdominal pain, and diarrhea were associated with high levels of breath hydrogen. CONCLUSION: The findings indicate that the Kuwaiti volunteers had higher breath hydrogen levels than Asians, but the incidence of lactose intolerance was similar in both groups. PMID- 12766334 TI - Retinoids and cancer: antitumoral effects of ATRA, 9-cis RA and the new retinoid IIF on the HL-60 leukemic cell line. AB - OBJECTIVE: To compare the antitumoral effects of all-trans retinoic acid (ATRA) and 9-cis retinoic acid (9-cis RA) with those of 5-OH,11-O-hydrophenanthrene (IIF), a new derivative of retinoic acid. MATERIALS AND METHODS: The effect of retinoids was tested on cell line HL-60. Cell differentiation and apoptosis were evaluated by morphological and biochemical analysis as BCL-2 protein and by DNA fragmentation assay. The ability to activate retinoic acid receptors (RAR) and/or retinoid X receptors (RXR) and to modulate gene expression was determined by transactivation assay. RESULTS: With cell line HL-60, the antiproliferative effect of IIF was stronger than that of ATRA and 9-cis RA. Following retinoid treatment, cells appeared to differentiate and apoptotic cells were observed. The appearance of DNA laddering and a decrease in the amount of BCL-2 protein confirmed apoptosis. IIF transcriptionally activated RXR-gamma more than RAR alpha. CONCLUSION: The findings indicate that IIF transcriptionally activates RXR gamma preferentially, induces apoptosis and has a more antiproliferative activity than ATRA and 9-cis RA on cell line HL-60. PMID- 12766335 TI - Acceptability for the use of postpartum intrauterine contraceptive devices: Assiut experience. AB - OBJECTIVES: To evaluate the acceptance of postpartum intrauterine contraceptive devices (PPIUCD) among the inhabitants of Assiut governorate, Egypt and to study the factors that influence this acceptance. SUBJECTS AND METHODS: Contraceptive counseling was given to 3,541 clients: 1,880 and 1,661 during the antenatal visits and postpartum hospitalization, respectively. Acceptors during antenatal counseling were to receive IUCDs via postplacental insertion in the case of vaginal delivery or transcesarean insertion in case of abdominal delivery. The clients who refused PPIUCD and chose interval IUCD insertion were referred to the Family Planning Clinic after the end of puerperium. Among postpartum counselees, PPIUCD acceptors received predischarge insertion within 48 h of delivery and the interval IUCD were referred to have IUCD inserted after the end of puerperium. The acceptance rate of both PPIUCD and interval IUCD and the percentage of actual insertions were recorded. The causes of both acceptance and refusal were also recorded. RESULTS: Of the 3,541 clients, 1,024 (28.9%) accepted the use of IUCD after delivery. Acceptance was approximately the same during antenal and postpartum counseling: 26.4 and 31.8%, respectively. Verbal acceptance was higher among women with formal education than among illiterate women. Planning another pregnancy in the near future, preference for another contraceptive method, namely lactational infertility, and complications from previous use of IUCD were the most common reasons for refusing the use of IUCD. Of the 1,024 verbal acceptors, only 243 (23.7%) had the actual insertion of IUCD. CONCLUSION: Both the acceptance and actual insertion of IUCD were low probably because the use of IUCD is a new concept in the community. For these women, the only opportunity to receive information about contraceptives is during childbirth when they are in contact with medical personnel. Hence, it is suggested that family planning should be integrated with maternal and child-care services in order to effectively promote the use of contraceptive devices in these women who otherwise would not seek the use of such a device. PMID- 12766336 TI - High prevalence of transfusion-transmitted virus infection in patients with chronic liver diseases in an endemic area of hepatitis B and C virus. AB - OBJECTIVE: To determine the prevalence and clinical impact of transfusion transmitted virus (TTV) DNA in patients with chronic liver diseases in the Southeast Anatolia region of Turkey where hepatitis B and C viral infections are endemic. SUBJECTS AND METHODS: Patients diagnosed with chronic liver disease by clinical, biochemical and histologic means were enrolled in the study. Serum samples of 60 patients (19 males, 41 females) with chronic liver diseases, and of 45 healthy volunteer blood donors as a control group were collected. The chronic liver disease group consisted of 11 patients with hepatitis B, 44 with hepatitis C and 5 with chronic liver disease of unknown etiology. Presence of TTV DNA was investigated by the polymerase chain reaction. Using a scoring system histological grading of inflammation and staging of fibrosis were performed only in the chronic hepatitis C group. RESULTS: TTV DNA was detected in 47 (78%) patients with chronic liver disease and 5 (11%) volunteers in the control group. The difference was statistically significant (p < 0.001). Ten of the 11 (91%) patients with hepatitis B, 32 of 44 (73%) of those with hepatitis C-related chronic liver disease, and 5 of 5 (100%) of the patients with cryptogenic liver disease were positive for TTV DNA. CONCLUSION: TTV is highly prevalent in patients with chronic liver diseases in Southeast Anatolia, Turkey but no pathogenic effect attributable to TTV infection was detected. PMID- 12766337 TI - Pain during mammography: impact of breast pathologies and demographic factors. AB - OBJECTIVES: The aim of this study was to systematically assess the pain experienced by patients undergoing mammography for various clinical presentations. SUBJECTS AND METHODS: Two hundred and twenty-five patients aged 25 85 years (45.43 +/- 8.25 years) presenting for mammography were included in the study. Presenting symptoms and clinical diagnosis were provided by the referring physicians and demographic information was obtained from self-reported questionnaires. Mammography results were recorded by the radiologist. Two different but reliable and valid measures of pain--Visual Analog Scale (VAS) and Pain/Discomfort Rating Scale (DRS)--were used to assess pain during mammography and data were statistically analyzed to examine the possible predictors of pain. RESULTS: Forty-nine percent of the patients reported pain during mammography when cut-off level of VAS score was 40; however, when the cut-off level was raised to 60 (considering the preexisting pain as presenting symptom in some patients) only 23% reported pain. With DRS, 7% reported pain, 27% discomfort and 66% neither pain nor discomfort. Biserial correlation between the VAS and DRS scores suggested strong positive agreement between the two measures of pain (r = 0.56, d.f. = 90, p < 0.01). Patients presenting with coexisting breast lumps and preexisting breast pain and those diagnosed with inflammatory conditions of the breast and fibrocystic changes experienced more pain during mammography. CONCLUSION: Preexisting breast pathologies and demographic factors such as age and educational level of the patient were important in reporting pain during mammography. This finding indicates that proper assessment of pain using standard measures and its association with breast pathologies and demographic factors is important for planning pain management in women undergoing mammography. PMID- 12766338 TI - A study of serological markers of hepatitis B and C viruses in Istanbul, Turkey. AB - OBJECTIVE: This study was undertaken to investigate the prevalence of HBsAg, anti HBs and anti-HCV positivity in Istanbul, Turkey. SUBJECTS AND METHODS: The frequencies of HBsAg, anti-HBs and anti-HCV positivity were determined in 1,157 randomly selected patients attending the outpatient clinic of Istanbul University Hospital, Istanbul, Turkey, during the years 1998 and 2001. All patients underwent complete physical and various routine laboratory examinations. RESULTS: Of the 1,157 patients, the prevalence of HBsAg, anti-HBs and anti-HCV was 6.6, 28.1 and 2.4%, respectively. It appeared that having dental and surgical procedures formed the risk factors for HBV infection. HBsAg positivity in the health care workers was not different from that of the other professions, but anti-HBs was significantly higher in this group. CONCLUSIONS: Our findings indicate that HBV infection occurs more frequently than HCV in Istanbul, and this poses an important health problem in the community. PMID- 12766339 TI - Influence of vitamin A consumption on resting metabolic rate and fasting respiratory quotient in severely obese subjects. AB - OBJECTIVE: To study whether or not the amount of vitamin A consumed affects the resting metabolic rate (RMR) and fat oxidation at rest in severely obese subjects. MATERIALS AND METHODS: In 239 obese subjects, RMR and fasting respiratory quotient (RQ) were determined by indirect calorimetry. Vitamin A consumption was calculated by the Czech PC program 'Nutrition'. The relation between the intake of vitamin A and RMR and RQ was tested by simple regression. High and low vitamin A consumers were defined by an upper and a lower quintile of vitamin A intake (>842 vs. <382 IU/day). RESULTS: The RMR for high and low vitamin A consumers were 7,693.5 +/- 1,557 and 7,479.8 +/- 1,708 kJ/day, respectively, while the corresponding values for fasting RQ were 0.800 +/- 0.077 and 0.809 +/- 0.049, respectively. No significant correlation was found between vitamin A consumption and both RMR and RQ. Similarly, there was no significant difference in RMR and RQ, as well as weight, body mass index, body fat, waist girth and food quotient between the two groups characterized by high and low consumption of vitamin A. However, the energy intake of high vitamin A consumers was significantly higher than that of low vitamin A consumers, due to higher carbohydrate and protein intake. CONCLUSION: There was no significant correlation between the vitamin A intake and RMR or RQ in obese subjects determined in this study. PMID- 12766340 TI - Maxillary canine-to-first premolar bilateral transposition in a female with Down syndrome. A case report. AB - OBJECTIVE AND IMPORTANCE: To report a case of the bilateral transposition of the maxillary canines with the premolars in a 17-year-old female with Down syndrome. CLINICAL PRESENTATION AND INTERVENTION: A 17-year-old female presented with moderate mental retardation; she was short and of small stature, and she had a round cherubic face and a broad forehead. Clinical intraoral examination revealed bilateral malpositions of the maxillary canines and premolars as transposition. She had an open mouth, macroglossia and a protruding tongue. The case required the combined orthodontic and prosthetic treatment to achieve an acceptable morphologic and esthetic occlusion. However, the parents refused to accept the proposed treatment. Extractions of the roots and caries treatment were done as these were considered priority for the patient. CONCLUSION: This rare and severe positional anomaly represents an orthodontic challenge and its correction involves treatment risk and requires a great deal of control and carefully applied mechanics. PMID- 12766341 TI - Anticonvulsant hypersensitivity syndrome: report of 2 cases from Kuwait. AB - OBJECTIVE: To illustrate the clinical features, laboratory findings, and management of anticonvulsant hypersensitivity syndrome (AHS), emphasizing the importance of recognizing its multiple clinical components and raising awareness of the cross-sensitivity among different anticonvulsants. CLINICAL PRESENTATION AND INTERVENTION: Two cases of AHS due to carbamazepine and a combination of sodium valproate and lamotrigene are reported. Both patients presented within the first month of starting the new antiepileptic medication with fever, skin rashes, hematological abnormalities, and hepatitis. The offending antiepileptic drugs were immediately stopped in both cases. Skin rashes responded to intravenous immunoglobulin in case 1 and to intravenous hydrocortisone in case 2. CONCLUSION: AHS is a serious, life-threatening condition. This report demonstrates that the most important steps in the management of AHS are to recognize the disorder, discontinue the offending antiepileptic drug, and provide supportive care in an inpatient setting and treat with benzodiazepines if seizures occur. PMID- 12766342 TI - Reversing transmembrane electron flow: the DsbD and DsbB protein families. AB - DsbD and DsbB are two proteins that in Escherichia coli catalyze transmembrane electron flow in opposite directions, thereby allowing reversible oxidoreduction of periplasmic dithiol/disulfide-containing proteins. We have identified all recognizable homologues of these two proteins in the databases and have conducted structural and phylogenetic analyses of the two families. The larger DsbD family is more diverse in sequence, topology, function and organismal distribution than the smaller DsbB family. DsbB homologues are rarely found outside of the proteobacteria, although DsbD homologues are found in many bacterial kingdoms as well as archaea and plant chloroplasts. Few organisms with a fully sequenced genome and a DsbB homologue lack a DsbD homologue, and most of these DsbD homologues fall within two clusters in the DsbD tree, exhibiting phylogenetic relationships that are the same as those observed for the DsbB proteins. These observations suggest that a subset of the DsbD homologues evolved in parallel with the DsbB family to perform a single unified function involving reversible extracytoplasmic protein dithiol-disulfide bond interchange. DsbD family proteins are shown to have arisen by an internal gene duplication event, and this observation leads to prediction of the pathway taken for the evolutionary appearance of the different protein topological types found within this family. PMID- 12766343 TI - Maize Activator transposase expressed in Saccharomyces cerevisiae from a genomic clone: detection via Elisa, and proposed use in complementation studies. AB - The maize Activator (Ac) transposase (TPase) was expressed as a histidine (His) tagged protein in Saccharomyces cerevisiae from a full-length genomic clone. Expression was demonstrated via the highly specific nickel-coated Elisa plate method, using an anti-His antibody and 2 separate anti-Ac TPase antibodies, to Ac residues 103-465 and 189-807. Ac TPase expression in S. cerevisiae is important for two reasons: (a) because the expression from a genomic clone herein permits the future study of RNA splicing mechanisms in common between maize and yeast systems, and (b) because a yeast system can easily be used for demonstrating complementation of function. Thus, such transformed yeast systems could be used in future to experimentally test whether Ac TPase could complement various yeast mutations. Specifically, Ac TPase may be able to complement (i.e., provide the same function) to yeast transcription factor mutants, or, to genes mutated in other essential yeast functions. If confirmed, this would lend support to Barbara McClintock's hypothesis that transposable elements can serve as 'controlling elements' within the genome, by their ability to supplement other essential genes' functions, as needed. Work herein is contrasted with existing studies on Ac in yeast. PMID- 12766344 TI - Non-Shine-Dalgarno initiators of translation selected from combinatorial DNA libraries. AB - In a search for non-Shine-Dalgarno (non-SD) translational initiators, two combinatorial expression libraries (denoted R(1) and R(2)) were constructed containing randomized decanucleotide regions placed at either 6 (R(1)) or 11 (R(2)) nucleotides upstream of a modified chloramphenicol acetyltransferase (CAT) gene. To prevent sporadic formation of SD-like sequences the content of G in the randomized region was restricted to 3% only. The two libraries were transformed in Escherichia coli cells and screened for chloramphenicol (Cm) resistance. More than 50 clones capable of tolerating Cm concentrations from 50 micro g/ml to more than 800 micro g/ml were selected. With few exceptions only, the non-SD sequences found in the Cm-resistant clones did not show any significant homology with other known non-SD initiators or enhancers of translation. Statistical (chi(2)) analysis of the distribution of nucleotides in the new non-SD translational initiators showed a different pattern from that of the conventional SD sequences. In few of the clones the yield of CAT exceeded that of the referent (SD containing) construct. The most productive clones carried the decanucleotides ATTTACCTCC, CCAATCTAC, TTCAATATTT, and TATTCCCCCA, and the corresponding yield of CAT obtained with them was 2.70, 2.06, 2.12 and 1.32 times, respectively, higher than that of the SD-bearing construct. PMID- 12766345 TI - Immobilization of Saccharomyces cerevisiae cells to protein G-Sepharose by cell wall engineering. AB - In this work, we explored the possibility of using the targeting of a heterologous protein to the cell wall of Saccharomyces cerevisiae, by fusing it to a cell wall protein, to construct yeast strains whose cells display on their surface proteins that bind to a matrix, so as to achieve the immobilization of the whole cells. With this aim, we created a gene fusion that comprises the region responsible for attachment of a cell wall protein to the cell wall, and the IgG binding region of staphylococcal protein A, and expressed it in the mnn1mnn9 strain of S. cerevisiae. The surface display of the protein A-Icwp fusion protein was positively monitored; however, direct immobilization of the cells on an IgG-Sepharose matrix did not produce the expected results, probably due to the fusion protein not being completely exposed on the surface of the cells. To solve this problem we incubated the cells first with rabbit preimmune serum and then with goat anti-rabbit IgGs, so as to create a complex (yeast cell protein A-rabbit IgG-goat IgG). Cells treated in this way were successfully immobilized on a protein G-Sepharose matrix, due to the binding properties of goat IgGs to streptococcal protein G. PMID- 12766346 TI - Early OspA immune complex formation in animal models of Lyme disease. AB - Infection with Borrelia burgdorferi, the cause of Lyme disease, has been accompanied by a puzzling delayed antibody (Ab) response to B. burgdorferi antigens (Ags) including the abundant organism-specific outer surface proteins, such as the 31-kD OspA. In humans the response to nonspecific B. burgdorferi Ags has required 3-6 weeks. The response to OspA has rarely been detected by conventional methodology until months after infection, despite demonstrable T cell reactivity. Tick inoculation and low-dose intradermal inoculation animal models have been characterized by a comparable response to OspA. Using more sensitive biotin-avidin immunoblots and immune complex (IC) dissociation techniques, we demonstrated in humans that Ab to OspA is formed early but may remain at low levels or bound in IC. To see if this was a universal biologic response, animal models were analyzed by these methods. The results with mice, monkeys and rabbits show that IC Ab to OspA may be detected at the onset of infection. The data suggest that these animal models may be used to understand the immune response to B. burgdorferi and the pathogenesis of Lyme disease. With attention to unique B. burgdorferi Ags, these results are likely to have both clinical and diagnostic importance. PMID- 12766347 TI - Phylogenomic analysis of the Giardia intestinalis transcarboxylase reveals multiple instances of domain fusion and fission in the evolution of biotin dependent enzymes. AB - Sequencing of the gene encoding a pyruvate carboxylase-like protein from the amitochondrial eukaryote Giardia intestinalis revealed a 1,338 aa protein composed of acetyl-CoA carboxyltransferase (ACCT), pyruvate carboxyltransferase (PycB), and biotin carboxyl carrier protein (BCCP) domains, linked in a single polypeptide chain. This particular domain combination has been previously seen only in the methylmalonyl-CoA:pyruvate transcarboxylase from Propionibacterium freudenreichii, where each of these domains is encoded by an individual gene and forms a separate subunit. To get an insight into the evolutionary origin and biochemical function of the G. intestinalis enzyme, we compared its domain composition to those of other biotin-dependent enzymes and performed a phylogenetic analysis of each of its domains. The results obtained indicate that: (1) evolution of the BCCP domain included several domain fusion events, leading to the ACCT-BCCP and PycB-BCCP domain combinations; (2) fusions of the PycB and BCCP domains in pyruvate carboxylases and oxaloacetate decarboxylases occurred on several independent occasions in different prokaryotic lineages, probably due to selective pressure towards co-expression of these genes, and (3) because newly sequenced biotin-dependent enzymes are often misannotated in sequence databases, their annotation as either carboxylases, decarboxylases, or transcarboxylases has to rely on detailed analysis of their domain composition, operon organization of the corresponding genes, gene content in the particular genome, and phylogenetic analysis. PMID- 12766348 TI - Radiation and functional specialization of the family-3 glycoside hydrolases. AB - A phylogenetic analysis of the glycoside hydrolases of family 3 (GH3s) was conducted in order to infer particular trends in its evolution: functional specialization, gene transfer events, gene duplications and paralogous evolution, and gene deletions. The phylogenetic analysis of GH3s revealed six clusters, i.e., A, B, C, D, E, and F that could fit the definition of 3 sub-families, i.e., AB, AB' and AB". While the sub-families AB' and AB" contain a single cluster, F and E, respectively, the AB sub-family is sub-divided into four clusters. Global analysis of the GH3 phylogenetic tree suggests a primary burst of amplification of the GH3s that might have led to these sub-families. Specializations, gene transfers, and gene duplications among each of these sub-families and phylogenetic clusters might then have occurred and have been inferred. The fine comparison of the enzyme properties and phylogenetic relationships of GH3s allowed to detect common functional groups that belong to the same cluster (D, E or F), or sub-cluster (A1, A2 or B2). The prokaryotic and eukaryotic beta xylosidases and beta-glucosidases belong to the AB and AB' sub-families, and the N-acetylglucosaminidases are in sub-family AB" (in cluster E). In some instances (B1, B2, C1, C2, and C3), the lack of data and/or the high heterogeneity of the hydrolytic properties did not allow to infer a particular link between an enzyme functional group and a phylogenetic cluster, suggesting the emergence of some highly specialized GH3s. PMID- 12766349 TI - Proliferation and hepatic differentiation of adult-derived progenitor cells. AB - Hepatic progenitor cells, capable of maturing into hepatocytes and biliary cells, are hypothesized to be involved in all forms of liver regeneration and may prove clinically useful at reconstituting damaged livers. A murine hepatic progenitor cell population from young adult liver tissue has been isolated and characterized to establish a model for the development of liver cell therapies and for analysis of immune responses after transplantation. Hepatic progenitor cells were isolated from 3- to 6-week-old C57BL/6 mice using modifications of a two-stage liver perfusion technique followed by low speed centrifugation. Cellular analysis by phase contrast, fluorescent and confocal microscopy demonstrated that the hepatic progenitors (1) formed ex vivo colonies with a morphological appearance similar to committed hepatocytic progenitors isolated from embryonic mice and rats; (2) they are smaller than mature hepatocytes; (3) in culture they demonstrated peak expression of an oval cell marker at day 14, whereas albumin expression continued to increase beyond day 21 of culture, and (4) a subset of the progenitors phenotypically differentiated into mature hepatocytes or biliary cells. The unique antigenic profile of these hepatic progenitor cells and their ability to differentiate suggests that purification of the cells should allow for their potential use in transplantation. PMID- 12766350 TI - Endometrial receptivity: selective adhesion competence of rabbit uterine epithelium for trophoblast but not for various tumor cells. AB - A modification of an established in vitro model for embryo implantation was used to probe the receptive uterine epithelium for any specificity of interaction with various invasive cells other than trophoblast. Endometrial explants consisting of stroma and epithelium taken from pseudopregnant rabbits were cultured in the presence of progesterone in order to regenerate a complete epithelial lining while maintaining the receptive state. Such precultured fragments were brought into contact with multicellular spheroids of different invasive tumor cell lines from different species. In contrast to the trophoblast of the rabbit blastocyst (previous publication), none of the tumor cell lines was able to adhere to intact epithelium of endometrial fragments nor to penetrate it. The uterine epithelium was also an insurmountable barrier for tumor cell spheroids confronted with the epithelium of fresh complex explants consisting of endometrium and myometrium or for spheroids introduced into the uterine lumen of pregnant/pseudopregnant rabbits at the periimplantation phase. However, all tumor cells were able to adhere to and mostly also to invade into the endometrial stroma when it was exposed artificially, i.e. when the epithelium was removed. These results suggest that the receptivity of rabbit uterine epithelium shows a remarkable selectivity with respect to cell type (trophoblast) and species (rabbit, not human, mouse, or rat). PMID- 12766351 TI - Embryonic blastemic changes in retinoic acid-induced hindlimb deformity. AB - In this study we analyzed, on an experimental model, blastemic changes occurring during the embryonic period that may later cause fetal hindlimb deformity. Experimental induction of clubfoot-like deformity in rat fetuses was performed by maternal administration of retinoic acid (RA) (120 mg/kg body weight) as a single intragastric dose on day 10 of pregnancy (previous phase of this assay). The caudal and hindlimb blastemic changes were studied by mitosis count and stereological, immunohistochemistry and AgNOR techniques in the hindlimbs of 15 day embryos and in the caudal somites of 11-day embryos. In 15-day embryos from the assay group, hypoplasia and misorientation of hindlimbs were present in 90% of the cases. The histological study showed a blastemal defect as follows: (a) reduction in mesenchymal cell activity (mitotic and AgNOR activities); (b) increase of volume of vascular lumen; (c) reduction in volume of nerve structures, and (d) reduction in the percentage of pre-rhabdomyoblastic cells. In 11-day embryos from the assay group, caudal somites showed disruption, including loss of usual morphology. Moreover, somitic AgNOR activity decreased compared to the control group. The greatest reduction in the number of black-dots per cell was in the myotome. These findings suggest that a certain pathology of the somites, very little studied to date, might be involved in clubfoot pathogenesis. PMID- 12766352 TI - Hyaluronan is differently located in arteries and veins. An immunohistochemical study in the rat. AB - The matrix components of the vessel wall are of great importance for the function of the vessel system of which both endurance and elasticity are prerequisites. One component of the vessel wall is the glycosaminoglycan hyaluronan (HYA) with its unique physico-chemical properties, e.g. viscoelasticity and barrier function. The present study aimed to map and compare the normal localisation and distribution of hyaluronan in the arteries and veins of both adult and newborn rats, using a specific staining method utilizing a hyaluronan-binding protein. The hyaluronan stained clearly different in veins and arteries both in newborn and adult rats. In the veins, the tunica intima stained intensely whereas the corresponding area in the arteries only showed a weak and scattered staining pattern. In the adult rats, the matrix between the smooth muscle cells of the tunica media of the veins had a clearly positive staining pattern compared to the media in the arteries which showed only a few scattered areas of positive staining. In the newborn rats, the media of the arteries stained more intensely. In both newborn and adult rats, the adventitia stained intensely both in veins and arteries. Moreover, the HYA-staining pattern differed in veins in different regions of the body. Since HYA is known to be involved in many different biological processes, the results are of importance to understand physiological properties of the vessel tree and to explain the development of vascular diseases. PMID- 12766353 TI - Changes in duodenal mast cells in response to dehydroleucodine. AB - INTRODUCTION: Dehydroleucodine (DhL), a sesquiterpene lactone isolated from Artemisia douglasiana Besser, prevents gastroduodenal damage elicited by necrosis inducing agents such as absolute ethanol. Changes in the number of mast cells or evidence of activation of the cells for mediator release have been observed in a wide spectrum of disease processes involving the gastrointestinal tract. In the present study we examined the effects of DhL on duodenal mast cell population and their histamine content, with the goal of throwing more light on the mechanism of action of the drug. MATERIALS AND METHODS: Male Rockland mice (n = 30) were divided into two groups and administered orally with 0.4% carboxymethylcellulose (CMC; control group) or DhL in 0.4% CMC, 40 mg/kg body weight (DhL group). The animals were killed 60 min after dosing and their duodena were removed. Mucosal and submucosal mast cells were studied by light and electron microscopy, and the duodenal histamine content was measured by high-performance liquid chromatography. RESULTS: DhL increased the number of mast cells in the submucosal layer. This was related to an increase in the tissue histamine levels (from 324 +/- 19.14 to 1,284 +/- 20 pg/mg tissue, in controls and DhL-treated, respectively). The mast cells in the submucosa from the control group showed a cytoplasm containing a predominant population of homogenously dense granules, and the DhL-treated group exhibited swollen granules showing different degrees of particulation. The mucosal mast cell population showed no modifications in response to the cytoprotective agent. CONCLUSIONS: DhL induces (1) a selective increase in the number of mast cells in the submucosal layer and (2) changes in the distribution and appearance of their secretory granules. These findings, probably associated with the higher histamine levels after DhL treatment, could be involved in the cytoprotective action of the lactone, previously reported by us. PMID- 12766355 TI - Transcranial color duplex sonography in cerebrovascular disease: a systematic review. AB - Transcranial color-coded duplex sonography (TCCS) enables the reliable assessment of intracranial stenoses (sensitivity 94-100%, specificity 99-100%), occlusions (middle cerebral artery: sensitivity 93-100%, specificity 98-100%) and cross-flow through the anterior (sensitivity 98%, specificity 100%) and posterior (sensitivity 84%, specificity 94%) communicating arteries without using potentially dangerous compression tests, as well as the midline shift in hemispheric infarcts. Ultrasound contrast agents (UCAs) increase the number of conclusive TCCS studies and allow the definite evaluation of intracranial arteries in most patients. TCCS is also useful for diagnosis and monitoring of vasospasm and detection of supratentorial hematomas (sensitivity 94%, specificity 95%) and aneurysms (sensitivity per patient 40-78%, specificity 90-91%), and may identify arteriovenous malformations. New developments are (1) UCAs that may increase the number of conclusive TCCS studies, (2) cerebral perfusion assessment, (3) measurement of arteriovenous cerebral transit time, which might enable the detection of small-vessel disease, and (4) site-targeted UCAs that may improve diagnosis and local drug delivery. PMID- 12766354 TI - Sleep deprivation inhibits expression of NADPH-d and NOS while activating microglia and astroglia in the rat hippocampus. AB - This study investigated the expression of nitric oxide (NO)-synthesizing enzymes and the glial reaction in the rat hippocampal formation following sleep deprivation for 5 days. Nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) reactivity was markedly reduced in the hippocampal CA1, CA2 and CA3 sectors as well as in the dentate gyrus, suggesting a suppression of NO production in these areas. Microglial cells were hypertrophic and showed an up regulation of complement type 3 receptors as determined by antibody OX-42. However, expression of major histocompatibility complex class I and II antigens, and antigen of monocyte/macrophage lineage marked by OX-18, OX-6 and ED1, respectively, was undetected. Astrocytes also displayed hypertrophied processes with enhanced glial fibrillary acidic protein (GFAP) immunoreactivity. Western blots of hippocampal tissues corroborated the above-mentioned morphological findings in that expression of NO-synthase (NOS) was decreased while that of OX 42 and GFAP was increased in the sleep-deprived rats. Since NO is thought to be involved in memory consolidation processes in the hippocampus during sleep, the inhibition of NADPH-d and NOS reactivities may account for the memory decline after long-term sleep deprivation. The concomitant reactions in microglia and astrocytes suggest the involvement of these cells in the deleterious effect of prolonged sleep deprivation. PMID- 12766356 TI - Time delay as a parameter for cerebrovascular reactivity in patients with severe carotid stenosis. AB - BACKGROUND: Amplitude responsiveness of cerebral blood flow velocity (CBFV) to CO(2) is an established test of cerebrovascular reserve capacity. We aimed to analyze the time course of CBFV during CO(2 )reactivity testing in patients with severe carotid stenosis. METHODS: 87 patients were assigned to three different groups according to the degree of stenosis (A 75-89%, B 90-94%, C 95-100%) and CBFV dynamics was assessed by transcranial Doppler sonography during hypercapnia induced by inhalation of 7% CO(2. )Time constants for the duration of CBFV and the end-tidal CO(2 )partial pressure (Pet(CO2)) increase and decrease were calculated. In addition, time delays between subsequent changes of Pet(CO2) and CBFV during on- and offset of hypercapnia were determined. RESULTS: There was a significant reduction of conventional CO(2) reactivity on the side ipsilateral to the stenosis in all groups. Time constants of CBFV showed no major differences between ipsi- and contralateral sides or between different groups. The off delay between the decrease in Pet(CO2) and the consequent decrease in CBFV was the only out of 6 different dynamic parameters which significantly differentiated critical stenosis and occlusion from severe stenosis. Retest variability showed a highly significant correlation. CONCLUSIONS: Analysis of the dynamic CBFV response during the applied CO(2) inhalation protocol revealed a significant and reproducible delay parameter which has power to detect cerebral hemodynamic compromise in patients with carotid artery stenosis similar to the conventional CO(2 )reactivity parameter. PMID- 12766357 TI - Public perception of stroke in Spain. AB - BACKGROUND AND PURPOSE: Stroke is one of the leading causes of death and disability in western countries. Improving attention to stroke demands better public information about the illness. The aim of the present study was to determine the current knowledge of stroke among the Spanish population. METHODS: We conducted a population-based telephone interview in Spain. A total of 3,000 respondents were selected at random following a systematic procedure aimed at filling the sex/age community quotas. RESULTS: A total of 16,016 telephone calls were made. Three thousand respondents were eligible and 2,884 completed the telephone interview. Only 4.5% of respondents knew the term stroke, 1,719 (59.6%) participants were able to mention at least one stroke risk factor, and only 940 (32.6%) could mention at least one stroke sign without prompting. Most respondents considered stroke to be a severe and dangerous illness that can be treated, but less than half of the respondents went immediately to a hospital when faced with a stroke event. Suitable stroke knowledge was only achieved by 302 (10.5%) respondents. Those who had home-based occupations and no previous stroke experience were less likely to have suitable stroke knowledge. CONCLUSIONS: This study confirms that our community is still unfamiliar with stroke. Wide population-based educational programs are needed to increase public knowledge of stroke. Our results indicate that people with home-based occupations should be the main target of our education campaigns. PMID- 12766358 TI - An East-West approach to the management of central post-stroke pain. AB - The development of neuropathic pain following stroke is not uncommon. The consequences include significant disabilities and depression. Treatment can often be difficult and responses unsatisfactory. We report a patient with severe central post-stroke pain (CPSP) of the right leg benefiting from a combination of Western multidisciplinary therapies AND acupuncture. A literature search has revealed that relatively few studies have been done on the management of CPSP, compared with other types of neuropathic pain. Amitriptyline and carbamazepine were found to produce positive effects on post-stroke pain in one small study; lamotrigine and gabapentin are two newer drugs which appear promising. To the best of our knowledge, the use of acupuncture for the treatment of CPSP has not been previously reported. PMID- 12766359 TI - The antiplatelet aggregation effects of aspirin suppositories. AB - To confirm that aspirin suppositories are an effective treatment for acute ischemic stroke, we examined the suppressive effects of 200-mg aspirin suppositories on platelet aggregation. Aspirin suppositories suppressed platelet aggregation induced by ADP or collagen, and the suppression continued for 24 h. There was no significant difference in suppression of platelet aggregation between aspirin administered by suppository and orally given aspirin. These results suggest that aspirin suppositories are a useful treatment for acute ischemic stroke. PMID- 12766360 TI - Noninvasive monitoring of cerebral oxygenation during vasomotor reactivity tests by a new near-infrared spectroscopy device. AB - BACKGROUND: Spatially resolved spectroscopy is a recently developed technique for noninvasive monitoring of cerebral tissue oxygenation using the photon diffusion theory. METHODS: We studied this technique with a new, commercial near-infrared spectroscopy (NIRS) device during vasomotor reactivity tests in 28 healthy volunteers (mean age 31.0 years; SD 10.6 years) and compared it with values assessed by the modified Beer-Lambert law and indices from simultaneous transcranial Doppler sonography of both middle cerebral arteries. We measured O(2) reactivity as percentage change of cerebral blood flow velocity (CBFV), as absolute change in the concentrations (measured in micromol/l) of oxygenated (HbO(2)), deoxygenated (Hb) and total hemoglobin (HbT), and as change in the tissue oxygenation index (TOI) during inhalation of 100% oxygen. CO(2) reactivity was calculated as percentage change of CBFV (NCR), as absolute change in the concentrations of HbO(2), Hb, and HbT (micromol/l), and as change in TOI (%) per 1% increase in end-tidal CO(2). RESULTS: One hundred percent oxygen inhalation lead to a decrease in CBFV (mean +/- SD: left -8.0 +/- 7.0%, p = 0.000; right 9.6 +/- 7.6%, p = 0.000), an increase in HbO(2) (0.99 +/- 1.07 micromol/l), Hbdiff (2.23 +/- 1.72 micromol/l), and TOI (3.1 +/- 1.5%), and a decrease in Hb ( 1.22 +/- 0.74 micromol/l), significant from baseline values (p = 0.0000). CO(2) reactivity was: NCR left 25.4 +/- 14.7%; NCR right 25.9 +/- 13.4%; HbO(2) 1.99 +/ 0.97 micromol/l; Hb -1.24 +/- 0.81 micromol/l; HbT 0.81 +/- 1.0 micromol/l, and TOI 3.7 +/- 2.2%. O(2) reactivity in TCD did not correlate with NIRS reactivities (Pearson p > 0.05), but NCR did correlate with changes in HbO(2), Hb, and TOI (Pearson p < 0.01). TOI was closely related to indices derived from the Beer Lambert law (Pearson p < 0.03), but not with mean arterial blood pressure or skin blood flow during vasomotor reactivity tests. CONCLUSION: Spatially resolved spectroscopy provides an encouraging, noninvasive new tool to study cerebral tissue oxygenation during vasomotor reactivity tests consistent with physiological changes. PMID- 12766361 TI - Contrast media effect on cerebral blood flow regulation after performance of cerebral or coronary angiography. AB - OBJECTIVE: The immediate hemodynamic effects of contrast agents are well documented. Less is known about their longer-lasting effects. We investigated the later effect of iopromide on the neurovascular coupling mechanism after the performance of cerebral or coronary angiography. Neurovascular coupling is a fine tuned and reliable mechanism adapting cerebral blood flow to cortical activity. When performing a visual stimulation and measuring the resultant flow velocity change in the posterior cerebral artery transcranial Doppler has been used to determine vascular integrity. METHODS: The visually evoked blood flow velocity response in the posterior cerebral artery was measured with transcranial Doppler before, 1-2 h after and the day after angiography. The overshoot of the flow velocity response and the stable flow velocity level at the end of the stimulation phase were used to compare the different conditions. RESULTS: A significantly diminished overshoot was found 1-2 h after angiography only in patients undergoing cerebral angiography. The stable blood flow velocity levels at the end of the stimulation phase remained unchanged in all cases. CONCLUSIONS: Our finding demonstrates effects of contrast media on dynamic blood flow regulation of the cerebral vasculature 1-2 h after application of direct contrast agent suggesting a possible concentration effect of the contrast agent. However, since the relative blood flow velocity increase under stable blood flow conditions remained constant, this dynamic alteration does not cause a reduction in cerebral blood flow. PMID- 12766362 TI - Effect of heart rate on regulative features of the cortical activity-flow coupling. AB - OBJECTIVE: Heart rate plays an important role in compensatory conditions of arterial pressure changes. Very little information, however, exists on its role in the dynamic adjustment of stimulated organ perfusion. We studied the influence of heart rate on the activity-flow coupling mechanism which adapts local cerebral blood flow in accordance with cortical activity. Since it does not affect heart rate or arterial blood pressure by itself, the commonly observed heart rate variability in test conditions was used to compare the flow response between different heart rate groups. For evaluation under stable heart rate conditions we performed a short test paradigm with a transcranial Doppler technique with the necessary high time resolution. METHODS: 168 healthy young volunteers (24 +/- 3 years of age) were grouped according to their heart rate in decade steps from 60 to 100 beats/min (mean: 82 +/- 9 beats/min). The visually evoked flow velocity responses in the posterior cerebral artery were evaluated according to a control system approach. Peak systolic and end diastolic data were evaluated separately. RESULTS: A correlation analysis between heart rate and baseline flow velocity as well as each of the control system parameters, i.e. gain, attenuation, rate time and natural frequency, revealed no significance. The flow responses did not differ among the heart rate groups as concluded from an ANOVA test. DISCUSSION: The increase in heart rate and the possible stress factors responsible for this seem to be of no relevance in regulative features of the activity-flow coupling. The almost identical time course of flow velocity responses among the groups showing a heart rate difference of up to 50% indicates an integrative principle in flow regulation supporting simplified concepts of flow adaptation. PMID- 12766363 TI - Doppler investigation of within-session reproducibility in a visual stimulation task to assess the volunteer-dependent variation. AB - OBJECTIVE: In functional magnetic resonance imaging (MRI) studies in humans the tightness of activity-flow coupling was questioned due to a notable within session variability of the signals obtained. To differentiate between moments of volunteer-dependent or technique-inherent variation in hemodynamic MRI measurements a method with a low technique-inherent variation is needed. Therefore, and because temporal features might also be of some relevance, we used the transcranial Doppler method. METHODS: In 22 healthy volunteers the flow velocity response to a visual stimulation task was evaluated. Conditions of 20 s of eye closure were altered with 40 s of silent reading, repeated 10 times. Averaged individual flow-velocity responses were evaluated from conditions of 40 s of stimulation and a 5-second time interval prior to the beginning of stimulation. The averaged data were subtracted from each single recording to calculate the mean squared differences (MSD). Statistical evaluation was performed according to a two-way ANOVA for repeated measures. To obtain additional qualitative data of the flow response, the individual flow curves were evaluated according to a control system approach specifying a second-order linear model. RESULTS: The coefficient of variation (COV) of the within-session MSD was 7% for the stimulation phase and 2.9% for the resting phase, whereas the respective COVs were 15 and 8.1% for the interindividual MSDs. The interindividual COVs of control system parameters ranged from 3.6 to 7.6%. The ANOVA resulted in a significant (p < 0.003) difference for the MSD values between stimulation conditions, whereas the test repetition (p = 0.55) and the test for an interaction of repetition and stimulation condition (p = 0.54) revealed no significance. DISCUSSION: Confirming the notion of a tight coupling between cortical activity and cerebral blood flow and being indicative of a low technique inherent variation, we found a high within-session reproducibility in a functional Doppler test. PMID- 12766364 TI - Cervical and intracranial atherosclerosis and silent brain infarction in Japanese patients with coronary artery disease. AB - PURPOSE: To investigate the prevalence of cervical and intracranial atherosclerosis and silent brain infarction in patients with coronary artery disease (CAD). METHODS: Cervical and intracranial atherosclerotic lesions on magnetic resonance angiography (MRA) and silent brain infarctions on magnetic resonance imaging (MRI) were investigated in comparison with the findings of coronary angiography in 133 consecutive patients with CAD. RESULTS: The mean severity scores of cervical and intracranial MRA lesions were significantly higher in the three-vessel CAD (0.40 and 0.53, respectively) than in the zero vessel CAD group (0.04 and 0.11). The mean scores of the maximal size and multiplicity of MRI lesion were also significantly greater in the two-vessel (1.00 and 1.44) and three-vessel CAD (0.94 and 1.26) than in the zero-vessel CAD group (0.27 and 0.50). The incidence of MRA lesion was markedly higher in patients with brain MRI lesion than in those without (51.1 vs. 6.5%). CONCLUSIONS: Serious coronary artery lesions were commonly accompanied by latent atherosclerotic lesions in the cervical and intracranial arteries besides silent brain infarction in patients with CAD. PMID- 12766366 TI - Low triglyceride, not low cholesterol concentration, independently predicts poor outcome following acute stroke. AB - BACKGROUND: Recent data have shown an unexpected association between poor outcome after acute stroke and lower serum cholesterol. Triglyceride concentration has been linked to coronary heart disease and stroke; however, there are currently no data on the relationship between triglyceride and stroke outcome. Such information may yield further mechanistic information on the relationship between lipids and stroke outcome. METHODS: We studied 1,310 nondiabetic patients presenting to our acute stroke unit with computed tomography-confirmed acute stroke. Fasting blood samples were drawn within 24 h of admission for glucose, lipids and a standard battery of biochemistry and hematological tests. Information on age, stroke type, admission blood pressure, smoking status, presence of atrial fibrillation, resolution time of symptoms and Oxfordshire Community Stroke Project clinical classification was collated. Stepwise proportional hazards regression was used to estimate the effect of the above variables on survival following stroke. RESULTS: Increased age (p < 0.0001), presence of atrial fibrillation (p = 0.009), hyperglycemia (p = 0.0005) and lower triglyceride level (p < 0.0001) independently predicted higher mortality; early resolution of symptoms (p = 0.005) independently predicted lower mortality. Although serum cholesterol level predicted outcome after adjusting for other prognostic factors, it did not remain significant when triglyceride level was entered into the model. The relative hazard per additional quartile of triglyceride was 0.84 (95% confidence interval 0.77-0.91). CONCLUSIONS: Low triglyceride concentration strongly predicts higher mortality following stroke, whereas serum cholesterol level is not an independent predictor. Outcome following stroke is thus related more strongly to triglyceride-rich than to cholesterol-rich lipoprotein concentrations. The mechanisms explaining this require further investigation. PMID- 12766365 TI - Serial changes in static and dynamic cerebral autoregulation after acute ischaemic stroke. AB - BACKGROUND: The longitudinal changes in static and dynamic cerebral autoregulation (CA) following acute ischaemic stroke are unknown and were assessed in this study. METHODS: Fifty-four ischaemic stroke patients were studied within 96 h of ischaemic stroke and again 7-14 days later, using transcranial Doppler techniques to measure CA. Results were compared to an age-, sex- and blood pressure (BP)-matched control group. Static BP pressor stimulus was produced by thigh cuff inflation and dynamic stimulus by rapid thigh cuff release. RESULTS: Dynamic, but not static, CA was globally impaired at initial (mean dynamic CA index 3.9 +/- 3.1 vs. 6.2 +/- 2.3, p < 0.005) and follow-up studies (dynamic CA 3.9 +/- 2.8 vs. 6.2 +/- 2.3, p < 0.01) in stroke patients compared to controls. Static CA was similar in stroke patients and controls and was unchanged during follow-up. CONCLUSIONS: Dynamic, but not static, CA is impaired after acute ischaemic stroke and remains abnormal for at least 1-2 weeks post ictus. These changes are present in both the affected and non-affected hemispheres and are unrelated to previous antihypertensive treatment, baseline BP levels or BP changes after stroke, age or stroke type. PMID- 12766367 TI - Prothrombin G20210A mutation, but not factor V Leiden, is a risk factor in patients with persistent foramen ovale and otherwise unexplained cerebral ischemia. AB - BACKGROUND: Paradoxical embolism via persistent foramen ovale (PFO) is suspected to be a frequent cause of stroke in younger patients. We investigated whether the prevalence of the risk factors for venous thrombosis factor V Leiden (FVL) and prothrombin G20210A mutation (PT G20210A) is increased in this group of patients. METHODS: We examined FVL and PT G20210A mutation in 220 patients (group 1) with cerebral ischemia associated with a PFO and without other etiology, in 196 patients with cerebral ischemia of an etiology other than PFO (group 2), and in 362 healthy subjects (group 3) from the same region in Germany. RESULTS: Heterozygosity for the PT G20210A mutation was more common in group 1 (5.0%) than in group 3 (1.4%; sex- and age-adjusted odds ratio 3.66; 95% CI 1.25-10.75; p = 0.01). By contrast, the mutation was not more common in group 2 (2.6%; odds ratio 1.50; 95% CI 0.42-5.41; p = 0.5). Prevalences of FVL were not different between groups. CONCLUSIONS: We identified PT G20210A but not FVL - the strongest genetic risk factor for deep venous thrombosis - to be significantly associated with stroke attributed to PFO. These findings rise doubts about the concept of paradoxical brain embolism as the dominating mechanism in stroke associated with PFO. PMID- 12766368 TI - The role of blood pressure in lacunar strokes preceded by TIAs. AB - Lacunar strokes (LS) are often preceded by repetitive transitory ischaemic attacks (TIAs) known as 'capsular warning syndrome'. The treatment of these symptoms remains controversial. Anticoagulants are often used in this situation, most of the time, however, with no or little benefit. Since it is not rare that these TIAs lead to a definite stroke, the therapeutic approach in the acute phase is very important. We report six patients presenting LS preceded by TIAs. All patients showed intermittent decrease of blood pressure (BP) coinciding with clinical worsening and leading to definite stroke in four of them. All were treated with full-dose intravenous heparin, which did not prevent further clinical deterioration. One patient received noradrenalin, which allowed stabilisation of the blood pressure values and complete resolution of the neurological symptoms. This observation suggests that in lacunar strokes preceded by TIAs monitoring and, if necessary, pharmacological increase of BP may prevent some patients from developing a definite stroke. Thus the 'capsular warning syndrome' could reflect a haemodynamic failure rather than repeated thrombo embolism within the lumen of a single perforating arteriole. PMID- 12766369 TI - No improvement of sustained attention after carotid endarterectomy as analysed by dark adaptometry. PMID- 12766370 TI - Midbrain ataxia: possible role of the pedunculopontine nucleus in human locomotion. PMID- 12766371 TI - Transient rotational vertigo as the initial symptom of a middle cerebral artery territory infarct involving the insula. PMID- 12766372 TI - Covered stent deployment and follow-up of a case of internal carotid artery pseudoaneurysm. PMID- 12766373 TI - Unilateral moyamoya disease in two brothers in the Netherlands. PMID- 12766374 TI - Achard-Levi syndrome: pupil-sparing oculomotor nerve palsy due to midbrain stroke. PMID- 12766376 TI - The role of therapeutic alliance in network therapy: a family and peer support based treatment for cocaine abuse. AB - The therapeutic alliance is a well-studied construct factor that is important to outcome in many forms of individual therapy. Therapeutic alliance has been rarely studied in group therapy and results in addiction treatment have been mixed. In this paper, we studied the presence of a therapeutic alliance in Network Therapy: an approach that uses peer and family support in addiction treatment. Twenty-one participants undergoing Network Therapy for cocaine addiction were observed on videotape, and were rated on therapeutic alliance using the Working Alliance Inventory and the Penn Helping Alliance Rating Scale. Results showed a significant positive correlation between therapeutic alliance and outcome as measured by the percentage of cocaine-free urine toxicology screens and by eight consecutive cocaine-free urines. PMID- 12766375 TI - Sexual identity and substance use among undergraduate students. AB - This study examined the association between sexual identity and use of alcohol and other drugs (AOD) among college undergraduate students. A survey regarding AOD use was administered to a random sample of 3607 undergraduate students. The sample included 65 self-identified lesbian or bisexual (LB) women and 54 self identified gay or bisexual (GB) men. Multivariate logistic regression indicated that while alcohol use did not differ for LB and heterosexual women, LB women were significantly more likely to experience certain AOD-related consequences, smoke cigarettes, and use marijuana, ecstasy, and other drugs. GB men were significantly less likely than heterosexual men to drink heavily but were more likely to use some drugs. These findings provide evidence that sexual identity is an important predictor of AOD use among undergraduate students. These findings support the need for continued research and intervention efforts that target LGB collegians. PMID- 12766377 TI - Smoking cessation, smoking reduction, and delayed quitting among smokers given nicotine patches and a self-help pamphlet. AB - Over-the-counter nicotine replacement raises questions regarding its "real world" efficacy. This was an open-label, prospective study of 223 smokers who received 42 free nicotine patches and a self-help booklet via shopping mall distribution. The overall quit rate 6 months following distribution of the nicotine patches was 22% (50/223), almost the same quit rate found 6 weeks following patch distribution (21%, 47/223). Twelve percent (27/223) were abstinent at both 6 weeks and 6 months. Among the 83 participants who did not quit, cigarettes smoked per day dropped from 28 to 18. A substantial subgroup of quitters (14%) who, although still smoking at 6 weeks, were smoke free at 6 months, and it appears they had purposefully delayed a serious quit attempt. These results support the usefulness of nicotine patches in helping smokers quit, even with only minimal intervention such as a self-help manual. PMID- 12766378 TI - Substance abuse and relationship violence among men court-referred to batterers' intervention programs. AB - There is extensive theoretical and empirical evidence linking substance abuse and marital violence. However in batterer populations, researchers have not compared hazardous and nonhazardous drinkers' substance use characteristics, marital aggression, extramarital violence (i.e., general violence), depressive symptomatology, marital satisfaction, and other relevant variables. In addition, much of the previous research on substance use and abuse in batterer populations employed a single, nonstandardized measure of substance use, and no previous research has examined substance use in court-mandated batterers' relationship partners. We recruited 150 men who were arrested for violence and court-referred to batterer intervention programs. We administered multiple measures of substance use and abuse and assessed the batterers' marital aggression, relationship satisfaction, depressive symptomatology, use of general violence, and their relationship partners' substance use. We also divided the sample into groups of Hazardous Drinkers (HD) and Nonhazardous Drinkers (NHD). Across the entire sample, half of the batterers had an alcohol-related diagnosis and approximately one third reported symptoms consistent with a drug-related diagnosis. Over one third of the total sample reported that their relationship partners were hazardous drinkers. Relative to the NHD group, the HD group scored significantly higher on measures of general violence, depressive symptomatology, alcohol use, alcohol problems, and drug problems. The HD group also reported significantly higher partner alcohol and drug use and abuse scores, relative to the NHD group. The results of the study suggest that substance use and abuse should routinely be assessed as part of batterer interventions and that batterer programs would be improved by offering adjunct or integrated alcohol treatment. PMID- 12766379 TI - Alcohol relapses associated with September 11, 2001: a case report. AB - The timing of the terrible events of September 11, 2001 (9-11), and an ongoing randomized clinical trial of case monitoring have allowed a prospective examination of the effects of trauma upon the relapse rates of a group of clients following alcohol detoxification. The clients studied in this report were enrolled in case monitoring prior to 9-11. Case monitoring consists of telephone contacts on a tapering schedule designed to help clients avoid relapses, reduce the severity of relapses that do occur, and get clients back into treatment, at less intense levels, than would occur without case monitoring. For those clients completing a telephone contact before and a telephone contact after 9-11, none of the clients drank between detox discharge and 9-11, while 42% drank by the first telephone contact after 9-11. Data from another study were analyzed and results counter the rival hypothesis that the case monitoring study results reflect an annual seasonal effect. Results suggest that terrorist events may lead to a greater likelihood of relapse for those in alcohol recovery. These effects may be ameliorated by public education and outreach. PMID- 12766380 TI - Gabapentin for the treatment of ethanol withdrawal. AB - Benzodiazepines (BZDs) are the drug of choice for the suppression of alcohol withdrawal symptoms. Gabapentin, a drug approved for use as adjunctive therapy in the treatment of partial seizures, has none of the BZD-type difficulties (drug interactions, abuse potential). We retrospectively report on the use of gabapentin for ethanol withdrawal in 49 patients. Thirty-one patients were treated in the outpatient program and 18 in the general inpatient psychiatric unit. Positive outcomes as evidenced by completion of gabapentin therapy were achieved in 25 out of 31 outpatients and 17 out of 18 inpatients. Statistical significance was reached regarding the positive relationship between prior ethanol use and inpatient "as needed" benzodiazepine use. Both sets of data suggest that gabapentin works well for the mild to moderate alcohol withdrawal patient. PMID- 12766381 TI - Specific drug delivery to the kidney. AB - The mesangial cells of the glomerulus, the proximal tubular cells and the interstitial fibroblasts are the first choice targets for renal drug delivery since they play a pivotal role in many disease processes in the kidney. In the present review, only targeting to the proximal tubular cell is addressed because only this has been studied extensively. Two approaches of drug delivery to the proximal tubular cell have been studied up to now, the prodrug/softdrug and low molecular-weight protein (LMPWP) approach. Most research on tubular specific drug delivery has focused on the development of amino-acid prodrugs that, after delivery, require activation by more or less kidney-selective enzymes. Large differences in renal selectivity are found. For some prodrugs, a rapid removal of the released drug from the kidney explained the low renal selectivity whereas for others, cleavage in non-target tissue and insufficient transport across the cell to the enzyme site seemed mainly responsible. The LMWP approach is based on drug attachment to a protein (<30 kD) that is freely filtered through the glomerulus and after accumulation is selectively catabolized in the lysosomes of the proximal tubular cell. Using LMWPs as drug carriers, a higher renal selectivity can be attained and a broader range of drugs can be attached while the rate of drug release can also be manipulated. The studies with captopril-lysozyme and naproxen-lysozyme clearly showed that targeting resulted in a higher renal selectivity and that drugs delivered into and regenerated in the proximal tubular cell exert renal selective pharmacological activity. Further testing will provide more definite data on the added value of this delivery technology. PMID- 12766382 TI - Cardiovascular therapy in patients with renal insufficiency. AB - Chronic renal failure and arterial hypertension run in parallel. New goal blood pressure levels have been established in 130/85 mmHg and 125/75 mmHg depending on the level of proteinuria being below or above 1 g/day. New and lower threshold BP (>130/85 mmHg) to initiate pharmacologic therapy are required in the presence of renal failure in order to facilitate the strict BP control that is required. Renal insufficiency is accompanied since its initial stages by a marked increase in cardiovascular risk and serum creatinine, its estimated clearance and the presence of proteinuria are very powerful predictors of a bad cardiovascular outcome. Hence, the need to consider that both renal and cardiovascular protection are obtained with such a strict BP control which, otherwise seems to require blockade of angiotensin II effects when proteinuria above 1g/day is present. Prevention of renal failure related to elevated blood pressure requires of strict blood pressure control, usually obtained with combination of two or more antihypertensive agents, one of them capable of blocking angiotensin II. Besides this, strict control of associated cardiovascular risk factors is also required. PMID- 12766383 TI - Treatment of hypertension in patients with renal disease. AB - Management of hypertension in people with kidney disease is challenging and generally requires at least three different and complementary acting antihypertensive agents to achieve the recommended blood pressure goal by the JNC VI and WHO guidelines of <130/85 mmHg. This is also true for the recent blood pressure goal for diabetes of <130/80 mmHg recommended by both the National Kidney Foundation and American Diabetes Association for reduction of cardiovascular risk and preservation of kidney function. Commonly used combinations include an ACE inhibitor, which has compelling indications for use in people with kidney disease with a diuretic, generally a thiazide type agent. Angiotensin receptor blockers have clearly shown effectiveness for slowing nephropathy progression in Type 2 diabetes and clearly have a role as first-line agents in that disease. If additional therapy is required, either a beta blocker or calcium antagonist may be added to this antihypertensive 'cocktail'. Beta blockers are particularly effective in people with a high sympathetic drive, i.e. high pulse rates, to lower pressure and reduce cardiovascular risk. Moreover, in recent studies their benefits on kidney function both by reducing proteinuria and slowing decline of kidney function make them good agents to add in the appropriate clinical setting. Given recent data from an analysis of the NHANES III database showing only 11% of people being treated for hypertension with diabetic kidney disease have achieved the blood pressure goal of <130/85 mmHg, it's no wonder the incidence of people starting dialysis continues to climb. Physicians need to work harder and educate patients on the importance of achieving these lower blood pressure guidelines. PMID- 12766384 TI - How to use nitrates. AB - Nitrates are old drugs, available in many formulations, that are effective in relieving symptoms in various CAD syndromes as well as congestive heart failure. Nitrate tolerance is the major problem limiting nitrate efficacy and use by physicians. Currently, an intermittent dosing strategy is the only practical way to limit the development of tolerance or attenuation of nitrate actions. PMID- 12766385 TI - Antioxidant effects and anti-elastase activity of the calcium antagonist nicardipine on activated human and rabbit neutrophils--a potential antiatherosclerotic property of calcium antagonists? AB - Activated neutrophils which produce certain proteases, such as elastase and reactive oxygen species (ROS) are involved in oxidative stress and inflammation. In the present study, we have shown that nicardipine, a calcium channel blocker, affects the release of elastase and superoxide anion radicals (O(2-)) in vitro during human and rabbit neutrophil respiratory bursts. The drug inhibited the release of elastase and O(2-) by fMLP (N-formyl-methionylleucin-phenylalaninin), calcium ionophore (A23187) and PMA (phorbol-myristate-acetate)-stimulated human and rabbit neutrophils. Besides the release of elastase, strongly inhibited in the fMLP and A23187 stimulated systems, nicardipine affected elastase and O(2-) in a dose-dependent manner. The corresponding 50% inhibitory concentration (IC(50)) of nicardipine for elastase, released in PMA-stimulated human and rabbit neutrophils, was 15.95 +/- 0.17 microM and 18.06 +/- 0.08 microM, respectively, whereas for O(2-), the IC(50) of nicardipine in PMA, fMLP and A23187-stimulated human and rabbit neutrophils was 55.41 +/- 0.09 microM and 58.43 +/- 0.03 microM, 45.21 +/- 0.13 microM and 37.19 +/- 0.53 microM, 33.54 +/- 0.09 microM and 30.54 +/- 0.29, respectively. The mechanisms underlying the inhibition of elastase and superoxide anion radicals by nicardipine appear related to an inhibiting effect on the mobilisation of cytosolic calcium and on activation of protein kinase C (PKC). These antioxidant and anti-elastasic activities contribute to the properties of nicardipine, as positive side effects of its antihypertensive activity and may be useful to prevent inflammatory disorders (tissue damage, oxidative injury) involved in the pathogenesis of hypertension. PMID- 12766386 TI - In vitro ultrasound augmented clot dissolution--what is the optimal timing of ultrasound application? AB - The mechanism of ultrasound augmentation of pharmacological thrombolysis is yet unknown. The goal of this study is to find the best timing regimen for in-vitro ultrasound augmented clot dissolution by streptokinase, heparin and their combination. Blood clots from 4 donors were cut into 200-400 mg sections and randomized to no treatment with ultrasound; pre-treatment with ultrasound (before immersion); early treatment with ultrasound; or late treatment with ultrasound. Clots were placed in tubes containing either saline; heparin; streptokinase or streptokinase +heparin. All groups showed significant weight reduction (p < 0.001). Using the one way ANOVA test, we showed that ultrasound application resulted in a significantly higher rate of clots dissolution (p < 0.05) than without ultrasound in all of the solutions tested. We found no statistically significant difference between the three ultrasound regimens tested. In conclusion, in our in-vitro model, no single ultrasound timing schedule was found to provide better clot dissolution than the other schedules. This finding may suggest an additive effect between the ultrasound and the different solutions rather than a synergistic effect. PMID- 12766387 TI - Efficacy of percutaneous intramyocardial injections using a nonfluoroscopic 3-D mapping based catheter system. AB - BACKGROUND: Percutaneous transendomyocardial injection with an injection catheter is a new drug delivery method for e.g. therapeutic angiogenesis. Little is known about the efficacy of this drug delivery technique. We studied efficiency and retention of transendomyocardial injections with a NOGA guided injection catheter system by using scintigraphy with radio-labeled model drugs. METHODS: Ten non ischemic landrace pigs were used. In each animal 2-3 transendomyocardial injections were performed using a 3-D mapping based catheter system called NOGA. As a model for proteins like angiogenic growth factors we used (99m)Tc labeled albumin and as a model for small particles like microspheres or adenovirus we used (99m)Tc labeled colloid albumin. Efficiency of the injections and retention of the transendomyocardial deposited substance were evaluated by a gamma camera during and after injection of 0.1 or 0.2 ml. RESULTS: All 29 injections showed scintigraphic proof of intramyocardial deposition. The average injection efficiency of all 29 injections was 26 +/- 23%. The average injection efficiency of 0.1 and 0.2 ml injections were 33 +/- 30% (n = 8) and 24 +/- 20% (n = 21), respectively (p = 0.33). Intramyocardial retention curves of albumin showed a rapid wash-out within the first 2 hours of the injection, whereas the retention of colloid albumin showed no decrease. In conclusion, transendomyocardial delivery of proteins or particles with an injection catheter show favorable efficiency rates, however the retention time of intramyocardial deposited small proteins like albumin is short. This may indicate the need for sustained release systems of angiogenic growth factors for intramyocardal injection in therapeutic angiogenesis. PMID- 12766388 TI - Efficacy of transdermal nitroglycerin patches evaluated by dipyridamole-induced myocardial ischemia in patients with coronary artery disease. Comparison between continuous and intermittent schedule. AB - Nitrate tolerance remains a problem despite an enormous number of studies on this phenomenon. The aim of the present study was to compare the efficacy of nitroglycerin transdermal patches intermittently or continuously administered to patients with myocardial ischemia evaluated by the echocardiography dipyridamole stress test. We prospectively studied 34 coronary patients with stable myocardial ischemia. A double-blind, randomized, crossover study technique was used. After a run-in period (1 week) they underwent a dipyridamole stress test to evaluate nitrate responsiveness and then were randomized to 1 week of transdermal nitroglycerine 20 mg/24 h (two patches of 10 mg/24) administered either intermittently or continuously for 1 week. During the following week all patients were given placebo. In the final week, therapy was crossed over the alternate regimen. No significant changes in heart rate, systolic and diastolic blood pressure and rate-pressure product were observed at basal conditions and at peak of dipyridamole infusion among patients after placebo run-in period, after acute, 1 week of intermittent and 1 week of continuous nitroglycerine administration. At peak of dipyridamole infusion after acute administration of nitrate we observed a significant decrease in wall motion score index with respect to placebo. This pattern was similar during intermittent, but not continuous, patch therapy (p < 0.001). Our results suggest that transdermal nitroglycerin patches are an effective anti-ischemic medication, in reducing transient myocardial ischemia induced by dipyridamole. This anti-ischemic effect is lost when an overnight nitrate free dose interval is not used. Moreover the dipyridamole echocardiographic stress test, besides evaluating myocardial ischemia-induced wall motion abnormalities, is adequate to assess both the efficacy and the tolerance of transdermal nitrate therapy. PMID- 12766389 TI - Clinical profile of eprosartan. AB - Angiotensin II (AII) receptor blockers offer an alternative means of blocking the renin-angiotensin-aldosterone system (RAAS) to angiotensin converting enzyme (ACE) inhibitors. Being highly selective for the AII receptor subtype AT(1), AII receptor blockers may avoid side-effects associated with ACE inhibitor treatment, such as cough. Eprosartan is a non-biphenyl, non-tetrazole competitive blocker that is chemically distinct from other AII receptor blockers, which may account for differences in its pharmacological properties. It induces dual blockade of AT(1) receptors both presynaptically and postsynaptically, reducing sympathetic nerve activity to a significantly greater degree than other AT(1) receptor blockers. At the recommended dose of 600 mg once daily, eprosartan effectively lowers blood pressure (BP) in hypertensive patients to a similar degree as seen with other AII receptor blockers and ACE inhibitors. However, a greater proportion of patients achieved adequate BP control compared with enalapril. When eprosartan is given in combination with hydrochlorothiazide (HCTZ), it provides a significantly greater BP reduction compared with eprosartan alone. Eprosartan has a side-effect profile that is similar to placebo and to other AII receptor blockers, but is better than that of enalapril because it lacks the propensity to cause dry cough. Eprosartan is not metabolized by the cytochrome P450 enzyme system, and so has no interaction with drugs that affect this system. Eprosartan completely reverses renal vasoconstriction induced by AII and may, therefore, have further applications in situations where stimulation of the RAAS is a problem. In summary, eprosartan, alone or in combination with HCTZ, provides an effective and well-tolerated approach to lowering BP in patients with all grades of hypertension. Further development of eprosartan may offer therapeutic opportunities that go far beyond the current recommendations. PMID- 12766390 TI - Correlated motions of successive amide N-H bonds in proteins. AB - New nuclear magnetic resonance (NMR) methods are described for the measurement of cross-correlation rates of zero- and double-quantum coherences involving two nitrogen nuclei belonging to successive amino acids in proteins and peptides. Rates due to the concerted fluctuations of two NH(N) dipole-dipole interactions and to the correlated modulations of two nitrogen chemical shift anisotropies have been obtained in a sample of doubly labeled Ubiquitin. Ambiguities in the determination of dihedral angles can be lifted by comparison of different rates. By defining a heuristic order parameter, experimental rates can be compared with those expected for a rigid molecule. The cross-correlation order parameter that can be derived from a model-free approach can be separated into structural and dynamic contributions. PMID- 12766391 TI - Chemical shift-dependent apparent scalar couplings: an alternative concept of chemical shift monitoring in multi-dimensional NMR experiments. AB - The paper presents an alternative technique for chemical shift monitoring in a multi-dimensional NMR experiment. The monitored chemical shift is coded in the line-shape of a cross-peak through an apparent residual scalar coupling active during an established evolution period or acquisition. The size of the apparent scalar coupling is manipulated with an off-resonance radio-frequency pulse in order to correlate the size of the coupling with the position of the additional chemical shift. The strength of this concept is that chemical shift information is added without an additional evolution period and accompanying polarization transfer periods. This concept was incorporated into the three-dimensional triple resonance experiment HNCA, adding the information of (1)H(alpha) chemical shifts. The experiment is called HNCA(coded)HA, since the chemical shift of (1)H(alpha) is coded in the line-shape of the cross-peak along the (13)C(alpha) dimension. PMID- 12766392 TI - A strategy to obtain backbone resonance assignments of deuterated proteins in the presence of incomplete amide 2H/1H back-exchange. AB - Replacement of non-exchangeable protons by deuterons has become a standard tool in structural studies of proteins on the order of 30-40 kDa to overcome problems arising from rapid (1)H and (13)C transverse relaxation. However, (1)H nuclei are required at exchangeable sites to maintain the benefits of proton detection. Protein expression in D(2)O-based media containing deuterated carbon sources yields protein deuterated in all positions. Subsequent D/H-exchange is commonly used to reintroduce protons in labile positions. Since this strategy may fail for large proteins with strongly inhibited exchange we propose to express the protein in fully deuterated algal lysate medium in 100% H(2)O. As a side-effect partial C(alpha) protonation occurs in a residue-type dependent manner. Samples obtained by this protocol are suitable for complementary (1)H(N)- and (1)H(alpha)-based triple resonance experiments allowing complete backbone resonance assignments in cases where back-exchange of amide protons is very slow after expression in D(2)O and refolding of chemically denatured protein is not feasible. This approach is explored using a 35-kDa protein as a test case. The degree of C(alpha) protonation of individual amino acids is determined quantitatively and transverse relaxation properties of (1)H(N) and (15)N nuclei of the partially deuterated protein are investigated and compared to the fully protonated and perdeuterated species. Based on the deviations of assigned chemical shifts from random coil values its solution secondary structure can be established. PMID- 12766393 TI - Preparation, characterization, and NMR spectroscopy of encapsulated proteins dissolved in low viscosity fluids. AB - Encapsulating a protein in a reverse micelle and dissolving it in a low-viscosity solvent can lower the rotational correlation time of a protein and thereby provides a novel strategy for studying proteins in a variety of contexts. The preparation of the sample is a key element in this approach and is guided by a number of competing parameters. Here we examine the applicability of several strategies for the preparation and characterization of encapsulated proteins dissolved in low viscosity fluids that are suitable for high performance NMR spectroscopy. Ubiquitin is used as a model system to explore various issues such as the homogeneity of the encapsulation, characterization of the hydrodynamic performance of reverse micelles containing protein molecules, and the effective pH of the water environment of the reverse micelle. PMID- 12766394 TI - Model selection for the interpretation of protein side chain methyl dynamics. AB - A number of different dynamics models are considered for fitting (13)C and (2)H side chain methyl relaxation rates. It is shown that in cases where nanosecond time scale dynamics are present the extended Lipari-Szabo model which is explicitly parameterized to include the effects of slow motions can produce wide distributions of fitting parameters even in cases where the errors are relatively small and large numbers of relaxation rates are considered. In contrast, fits of (15)N backbone dynamics using this model are far more robust. The origin of this difference is analyzed and can be explained by the different functional forms of the spectral density in these two cases. The utility of a number of models for the analysis of methyl side chain dynamics is presented. PMID- 12766396 TI - Fast multi-dimensional NMR of proteins. AB - Three-dimensional HNCO and HNCA subspectra from a small protein (agitoxin, 4 kDa, enriched in carbon-13 and nitrogen-15), have been obtained by direct frequency domain excitation of selected carbon and nitrogen sites. This new technique applies an array of several simultaneous soft radiofrequency spin-inversion pulses, encoded (on or off) according to nested Hadamard matrices, and the resulting responses are extracted by reference to the same matrices. This not only simplifies the spectra but, by avoiding extensive sampling in the traditional evolution dimensions, affords a speed advantage of more than two orders of magnitude. PMID- 12766395 TI - A novel medium for expression of proteins selectively labeled with 15N-amino acids in Spodoptera frugiperda (Sf9) insect cells. AB - Whereas bacterial expression systems are widely used for production of uniformly or selectively (15)N-labeled proteins the usage of the baculovirus expression system for labeling is limited to very few examples in the literature. Here we present the complete formulations of the two insect media, IML406 and 455, for the high-yield production of selectively (15)N-labeled proteins in insect cells. The quantities of (15)N-amino acids utilized in the production of labeled GST were similar in the case of bacterial and viral expression. For the most studied amino acids essential for insect cells the (15)N-HSQC spectra, recorded with GST labeled in insect cells, showed no cross labeling and provided therefore spectra of better quality compared to NMR spectra of GST expressed in E. coli. Also in the case of amino acids not essential for Sf9 cells we were able to label a defined number of amino acid species. Therefore the selective labeling using the baculovirus expression vector system represents a complement or even an alternative to the bacterial expression system. Based on these findings we can provide a first simple overview of the network of the amino acid metabolism in E. coli and insect cells focused on nitrogen. For some amino acids the expression of labeled proteins in insect cells can replace the cell-free protein expression. PMID- 12766397 TI - NMR assignment and structural characterization of the fatty acid binding protein from the flight muscle of Locusta migratoria. PMID- 12766398 TI - Biosynthesis of isotopically labeled gramicidins and tyrocidins by Bacillus brevis. AB - The three-dimensional structure of bilayer-associated gramicidin A is available from a structural data base. This and related peptides are, therefore, ideal model compounds to use during the implementation and development of new NMR techniques for the structural investigations of membrane proteins. As these methods rely on the isotopic labelling of single, selected or all sites, we have, investigated and optimised biochemical protocols using different strains of the Gram-positive bacterium Bacillus brevis. With newly developed schemes for isotopic labelling large amounts of gramicidin and tyrocidin enriched with stable isotopes such as (15)N or (15)N/(13)C have been obtained at low cost. A variety of analytical and spectroscopic techniques, including HPLC, mass spectrometry and NMR spectroscopy are used to characterise the resulting products. PMID- 12766399 TI - Broadband RFDR with adiabatic inversion pulses. AB - With a view to obtain (13)C chemical shift correlation spectra of uniformly labelled peptides/proteins at high magnetic fields and high magic angle spinning frequencies (omega(r)/2pi or =4 kcal mol(-1). At -74 degrees C, the slow, large amplitude motion was best characterized as a two-site jump with a correlation time on the millisecond time scale, whereas at -60 degrees C it was diffusive on the microsecond time scale. The slow, large-amplitude motions of the C2'-H2' bond are most likely from interconversions between C2'-endo and C3'-endo by way of the O4'-endo conformation, whereas the fast, small-amplitude motions are probably librations of the C2'-H2' bond within the C2'-endo and C3'-endo potential energy minima. PMID- 12766420 TI - Cross-correlation suppressed T1 and NOE experiments for protein side-chain 13CH2 groups. AB - Relaxation measurements of side-chain 13CH2-groups of uniformly 13C labeled human ubiquitin were performed at 600 MHz and 800 MHz magnetic field strength at 30 degrees C. Dipole-dipole cross-correlated relaxation effects in T1 experiments were suppressed by the combination of radio-frequency pulses and pulsed field gradients during the relaxation delay leading to monoexponential relaxation decays that allow a more accurate extraction of the 13C T1 relaxation times. Heteronuclear [1H]-13C NOEs obtained by using different proton saturation schemes indicate that the influence of cross-correlation is small. The experimental T1 and NOE data were interpreted in a model-free way in terms of a generalized order parameter and an internal correlation time. PMID- 12766419 TI - Rapid and accurate calculation of protein 1H, 13C and 15N chemical shifts. AB - A computer program (SHIFTX) is described which rapidly and accurately calculates the diamagnetic 1H, 13C and 15N chemical shifts of both backbone and sidechain atoms in proteins. The program uses a hybrid predictive approach that employs pre calculated, empirically derived chemical shift hypersurfaces in combination with classical or semi-classical equations (for ring current, electric field, hydrogen bond and solvent effects) to calculate 1H, 13C and 15N chemical shifts from atomic coordinates. The chemical shift hypersurfaces capture dihedral angle, sidechain orientation, secondary structure and nearest neighbor effects that cannot easily be translated to analytical formulae or predicted via classical means. The chemical shift hypersurfaces were generated using a database of IUPAC referenced protein chemical shifts--RefDB (Zhang et al., 2003), and a corresponding set of high resolution (<2.1 A) X-ray structures. Data mining techniques were used to extract the largest pairwise contributors (from a list of approximately 20 derived geometric, sequential and structural parameters) to generate the necessary hypersurfaces. SHIFTX is rapid (<1 CPU second for a complete shift calculation of 100 residues) and accurate. Overall, the program was able to attain a correlation coefficient (r) between observed and calculated shifts of 0.911 (1Halpha), 0.980 (13Calpha), 0.996 (13Cbeta), 0.863 (13CO), 0.909 (15N), 0.741 (1HN), and 0.907 (sidechain 1H) with RMS errors of 0.23, 0.98, 1.10, 1.16, 2.43, 0.49, and 0.30 ppm, respectively on test data sets. We further show that the agreement between observed and SHIFTX calculated chemical shifts can be an extremely sensitive measure of the quality of protein structures. Our results suggest that if NMR-derived structures could be refined using heteronuclear chemical shifts calculated by SHIFTX, their precision could approach that of the highest resolution X-ray structures. SHIFTX is freely available as a web server at http://redpoll.pharmacy.ualberta.ca. PMID- 12766424 TI - 1H, 13C and 15N resonance assignments of a viral SET domain histone lysine methyltransferase. PMID- 12766423 TI - 1H, 13C and 15N backbone resonance assignments of the N-terminal domain of Drosophila GCM protein. PMID- 12766422 TI - Similarities between intra- and intermolecular hydrogen bonds in RNA kissing complexes found by means of cross-correlated relaxation. AB - The bond lengths and dynamics of intra- and intermolecular hydrogen bonds in an RNA kissing complex have been characterized by determining the NMR relaxation rates of various double- and triple-quantum coherences that involve an imino proton and two neighboring nitrogen-15 nuclei belonging to opposite bases. New experiments allow one to determine the chemical shift anisotropy of the imino protons. The bond lengths derived from dipolar relaxation and the lack of modulations of the nitrogen chemical shifts indicate that the intermolecular hydrogen bonds which hold the kissing complex together are very similar to the intramolecular hydrogen bonds in the double-stranded stem of the RNA. PMID- 12766425 TI - 1H, 15N and 13C backbone assignment of the green fluorescent protein (GFP). PMID- 12766427 TI - Simulations of voltage clamping poorly space-clamped voltage-dependent conductances in a uniform cylindrical neurite. AB - Significant error is made by using a point voltage clamp to measure active ionic current properties in poorly space-clamped cells. This can even occur when there are no obvious signs of poor spatial control. We evaluated this error for experiments that employ an isochronal I(V) approach to analyzing clamp currents. Simulated voltage clamp experiments were run on a model neuron having a uniform distribution of a single voltage-gated inactivating ionic current channel along an elongate, but electrotonically compact, process. Isochronal Boltzmann I(V) and kinetic parameter values obtained by fitting the Hodgkin-Huxley equations to the clamp currents were compared with the values originally set in the model. Good fits were obtained for both inward and outward currents for moderate channel densities. Most parameter errors increased with conductance density. The activation rate parameters were more sensitive to poor space clamp than the I(V) parameters. Large errors can occur despite "normal"-looking clamp curves. PMID- 12766428 TI - Computational constraints that may have favoured the lamination of sensory cortex. AB - At the transition from early reptilian ancestors to primordial mammals, the areas of sensory cortex that process topographic modalities acquire the laminar structure of isocortex. A prominent step in lamination is granulation, whereby the formerly unique principal layer of pyramidal cells is split by the insertion of a new layer of excitatory, but intrinsic, granule cells, layer IV. I consider the hypothesis that granulation, and the differentiation between supra- and infra granular pyramidal layers, may be advantageous to support fine topography in their sensory maps. Fine topography implies a generic distinction between "where" information, explicitly mapped on the cortical sheet, and "what" information, represented in a distributed fashion as a distinct firing pattern across neurons. These patterns can be stored on recurrent collaterals in the cortex, and such memory can help substantially in the analysis of current sensory input. The simulation of a simplified network model demonstrates that a non-laminated patch of cortex must compromise between transmitting "where" information or retrieving "what" information. The simulation of a modified model including differentiation of a granular layer shows a modest but significant quantitative advantage, expressed as a less severe trade-off between "what" and "where". The further connectivity differentiation between infra-granular and supra-granular pyramidal layers is shown to match the mix of "what" and "where" information optimal for their respective target structures. PMID- 12766426 TI - Tuning neocortical pyramidal neurons between integrators and coincidence detectors. AB - Do cortical neurons operate as integrators or as coincidence detectors? Despite the importance of this question, no definite answer has been given yet, because each of these two views can find its own experimental support. Here we investigated this question using models of morphologically-reconstructed neocortical pyramidal neurons under in vivo like conditions. In agreement with experiments we find that the cell is capable of operating in a continuum between coincidence detection and temporal integration, depending on the characteristics of the synaptic inputs. Moreover, the presence of synaptic background activity at a level comparable to intracellular measurements in vivo can modulate the operating mode of the cell, and act as a switch between temporal integration and coincidence detection. These results suggest that background activity can be viewed as an important determinant of the integrative mode of pyramidal neurons. Thus, background activity not only sharpens cortical responses but it can also be used to tune an entire network between integration and coincidence detection modes. PMID- 12766429 TI - Dynamics of spiking neurons connected by both inhibitory and electrical coupling. AB - We study the dynamics of a pair of intrinsically oscillating leaky integrate-and fire neurons (identical and noise-free) connected by combinations of electrical and inhibitory coupling. We use the theory of weakly coupled oscillators to examine how synchronization patterns are influenced by cellular properties (intrinsic frequency and the strength of spikes) and coupling parameters (speed of synapses and coupling strengths). We find that, when inhibitory synapses are fast and the electrotonic effect of the suprathreshold portion of the spike is large, increasing the strength of weak electrical coupling promotes synchrony. Conversely, when inhibitory synapses are slow and the electrotonic effect of the suprathreshold portion of the spike is small, increasing the strength of weak electrical coupling promotes antisynchrony (see Fig. 10). Furthermore, our results indicate that, given a fixed total coupling strength, either electrical coupling alone or inhibition alone is better at enhancing neural synchrony than a combination of electrical and inhibitory coupling. We also show that these results extend to moderate coupling strengths. PMID- 12766431 TI - Type I burst excitability. AB - We introduce the concept of "type I burst excitability", which is a generalization of the "normal" excitability that is well-known in cardiac and neural systems. We demonstrate this type of burst excitability in a specific model system, a pyramidal cell from the electrosensory lateral line lobe of the weakly electric fish Apteronotus leptorhynchus. As depolarizing current is increased, a saddle-node bifurcation of periodic orbits occurs, which separates tonic and burst activity. This bifurcation is responsible for the excitable nature of the system, and is the basis for the "type I" designation. We verify the existence of this transition from in vitro recordings of a number of actual pyramidal cells. A scaling relationship between the magnitude and duration of a current pulse required to induce a burst is derived. We also observe this type of burst excitability and the scaling relationships in a multicompartmental model that is driven by realistic stochastic synaptic inputs mimicking sensory input. We conclude by discussing the relevance of burst excitability to communication between weakly electric fish. PMID- 12766432 TI - 20 Questions of Gamblers Anonymous: A Psychometric Study with Population of Spain. AB - The authors present a psychometric study of the questionnaire proposed by the Gamblers Anonymous organization for the self-assessment of problem gambling. The study was carried out in Spain with two samples: one of 127 problem gamblers (mean age: 41.67; sex: 78.7% men; mean duration of the problem: 5 years) and the other of 142 social gamblers (mean age: 38.46; sex: 60% men). The questionnaire was self-applied, in the presence of one of the investigators in case any doubts arose. The results indicate that it is a good screening instrument, with high reliability (Cronbach's alpha = 0.94), good covergent validity (the correlation with the SOGS questionnaire is high (r = 0.94; p < 0.001)), good factorial validity (the questionnaire's structure is unidimensional and coherent (just one factor explains more than 50% of the variance)), and its discriminative power is high (diagnostic efficacy = 98.88%). The study offers a comparative analysis of this instrument with other measures of problem gambling, emphasizing its extraordinary performance. PMID- 12766430 TI - No parallel fiber volleys in the cerebellar cortex: evidence from cross correlation analysis between Purkinje cells in a computer model and in recordings from anesthetized rats. AB - Purkinje cells aligned on the medio-lateral axis share a large proportion of their approximately 175,000 parallel fiber inputs. This arrangement has led to the hypothesis that movement timing is coded in the cerebellum by beams of synchronously active parallel fibers. In computer simulations I show that such synchronous activation leads to a narrow spike cross-correlation between pairs of Purkinje cells. This peak was completely absent when shared parallel fiber input was active in an asynchronous mode. To determine the presence of synchronous parallel fiber beams in vivo I recorded from pairs of Purkinje cells in crus IIa of anesthetized rats. I found a complete absence of precise spike synchronization, even when both cells were strongly modulated in their spike rate by trains of air-puff stimuli to the face. These results indicate that Purkinje cell spiking is not controlled by volleys of synchronous parallel fiber inputs in the conditions examined. Instead, the data support a model by which granule cells primarily control Purkinje cell spiking via dynamic population rate changes. PMID- 12766434 TI - Personality Disorders Among Pathological Gamblers. AB - The objective of this study was to investigate the prevalence of DSM-III-R diagnostic categories of personality disorders in pathological gamblers and to highlight the possible association between such disorders, psychological distress and selected forms of gambling. The Personality Disorders Questionnaire-Revised and a battery of psychometric measures were administered to a sample of 82 consecutive admissions to a behavioral treatment program for gambling problems at an impulse control disorders research unit in Sydney, Australia. Seventy-three percent of subjects were male. The total sample reported having gambled a mean of 15 years of which, on average, the last 6.4 years were associated with problems. Results indicated that the majority of subjects met diagnostic criteria for at least one Personality Disorder (93%), with an average of 4.6 personality disorders per subject. The majority of gamblers evidenced personality disorders from the Cluster B grouping with particularly high rates of borderline, histrionic, and narcissistic personality disorders which were found to be associated with high levels of impulsivity and affective instability. Antisocial personality disorder and narcissistic personality disorder were both found to be possible mediators of the severity of the problem gambling behaviours. PMID- 12766433 TI - An Empirical Examinination of Jacobs' General Theory of Addictions: Do Adolescent Gamblers Fit the Theory? AB - The present study examines 817 adolescent High School students' gambling behavior. Participants completed the DSM-IV-J, a gambling screen examining severity of gambling problems along with the High School Personality Questionnaire (HSPQ), the Reynolds Adolescent Depression Scale (RADS) and a questionnaire devised by the authors inquiring about gambling behavior, substance abuse, alcohol consumption, and cigarette smoking. The study seeks to test the Jacobs' (1986) General Theory of Addictions, using problem and pathological adolescent gamblers as the prototype to test the model. The results obtained through path analysis and logistic regression suggest that Jacobs' theory is a plausible and likely explanation for the development of a gambling addiction amongst adolescents. The clinical implications are addressed. PMID- 12766435 TI - Response to Shaffer (1996): The Case for a 'Complex Systems' Conceptualisation of Addiction. AB - This paper responds to some of the issues raised by Shaffer's (1996) paper, "Understanding the Means and Objects of Addiction." The authors agree with, and in many respects support, the assertions made by Shaffer with respect to the conceptualisation of the problem of addiction. In particular, that it is the relationship of the addicted person with the object of their behaviour that defines addiction. However, it is argued that in addition to providing an operational definition for addiction, it is important to provide an explicit framework for the development of insight into the 'synergistic nature' of the multiple factors affecting the addiction process. That is, it is necessary to account for both the commonalities and the differences found across the spectrum of addictive behaviours. It is proposed that this might be provided by a 'complex systems' approach to the field of addiction. PMID- 12766436 TI - Parents' Attitudes and Knowledge Regarding Gambling Among Youths. AB - This study evaluates attitudes and knowledge of parents regarding gambling behaviors among youths, aged 5 to 17 years. Telephone interviews were conducted among 279 randomly selected parents (32% fathers and 68% mothers) in the Quebec City region. Results indicate that parents overestimate the age of children's first wagers and underestimate the probability that their own child has already gambled. Most parents (86%) believe that the availability of gambling for youths should be reduced and that schools should include prevention programs concerning problem gambling. Results also show that parents fail to associate excessive gambling with poor grades or with alcohol and drug use. Finally, 84% of the parents reported that they would accept buying lottery tickets for their child. These results suggest that prevention programs for excessive gambling among children should include information for parents. PMID- 12766438 TI - The Cognitive Psychology of Lottery Gambling: A Theoretical Review. AB - Despite the current popularity of the UK National, psychologists have tended to neglect lottery play. This review provides a summary of current research findings and outlines the main cognitive theories of gambling as related to non pathological lottery play. A discussion of various biases and irrational thinking patterns typically found in lottery gambling will be given. These will include the misunderstanding of lottery odds, a susceptibility to the gambler's fallacy and cognitive entrapment, a belief in hot and cold numbers, unrealistic optimism, a belief in personal luck, superstitious thinking, the illusion of control, the erroneous perception of near misses, a susceptibility to prize size and rollover effects, the framing of gambling outcomes and finally, the influence of social factors on lottery play. It is concluded that the psychology of lottery play needs a more unified theory which whilst largely cognitive in emphasis, should also incorporate social motivations such as those inherent in syndicate based lottery play. PMID- 12766439 TI - An Analysis of the Relationship of Alcohol to Casino Gambling Among College Students. AB - Research has found significant overlap in the problem drinker and pathological gambler populations. This finding leads to the question of whether the pairing of drinking and gambling at lower levels of intensity is similarly related to a variety of negative consequences. The data for the present study were gathered in Memphis, TN, and Reno, NV, from questionnaires completed by 835 students in two universities. The data indicate that about one-fourth of students who gamble in casinos frequendy or always drink while gambling. Drinking when gambling is significantly related for males, but not for females, to size of bet, obtaining additional money while at the casino, and losing more than one can afford. The analysis suggests that an increased effort should be made to inform even casual drinkers and casual gamblers of the dangers of pairing these behaviors. PMID- 12766437 TI - A Case Series of 44 Completed Gambling-Related Suicides. AB - This paper presents an analysis of 44 case records of suicide occurring between 1990 and 1997 in the State of Victoria, Australia, in which the State Coroner identified the presence of a putative gambling problem. Analysis of demographic data revealed that the majority of suicidal gamblers were male with a mean age of 40 years with 84% of the sample being either unemployed or from lower socioeconomic backgrounds. The most common method of suicide was carbon monoxide poisoning. A quarter of the victims left a suicide note. Evidence was found indicating that almost a third (31.8%) of cases had previously attempted suicide and one in four had sought some form of mental health assistance for their gambling problem. A number of putative risk factors were identified including comorbid depression, large financial debts and relationship difficulties. The relationship between crime, suicide and gambling and gender differences among suicidal gamblers was also examined. The authors conclude that further research is required to substantiate these initial findings which are based on retrospective accounts and secondary sources of evidence. PMID- 12766440 TI - Deregulation and Cross-Border Substitution in Iowa's Riverboat Gambling Industry. AB - When riverboat casinos began operating in Iowa in April 1991 a strict regulatory structure, consisting of mandated sailing, wagering and loss limits, and casino space restrictions, was imposed. The restrictions were removed in May 1994 after Illinois introduced riverboat gambling without space, betting, or loss limits. Using data on casino win and admissions in Iowa and Illinois, this study analyzes the impact that deregulation had on casino win, win per admission, and total admissions. The findings reveal that deregulation significantly increased casino win, win per admission, and total admissions in Iowa. Moreover, when focusing on the "Quad Cities" market, which encompasses both Iowa and Illinois casinos, deregulation is found to result in significant cross-border substitution. In addition, there is evidence that deregulation has partly resulted in existing gamblers losing more money as opposed to increasing the number of gamblers. PMID- 12766441 TI - Outcome Expectancies for Gambling: Empirical Modeling of a Memory Network in Federal Prison Inmates. AB - Outcome expectancies for gambling were explored in a group of 316 male medium security federal inmates with the multidimensional scaling statistical technique. Sixty-six possible outcomes for gambling were reduced to 16 after a factor analysis determined that 75% of the total variance was accounted for by the first 16 factors (eigenvalues > 0.75). The highest loading item for each factor was utilized in all subsequent analyses. Multidimensional scaling of these 16 items revealed the presence of two primary dimensions (positive-negative and arousing sedating), corroborating previous research on alcohol outcome expectancies (Rather et al., 1992) and general affective response (Russell, 1980). Significant group (non-gambling, non-problem gambling, possible problem gambling, probable pathological gambling) differences were observed on 12 of the 16 items and on 3 of 4 composite scales (positive expectancies, negative expectancies, arousing expectancies, sedating expectancies). PMID- 12766443 TI - Analysis of the National Gambling Impact Study Commission Act. AB - The following article by Keith Whyte represents the first of occasionally reprinted publications that are relevant to important historical events in the field of gambling studies. This article describes the act that led to the formation of the National Gambling Impact Study Commission. This Commission released its findings to the President and Congress of the United States during June of 1999. PMID- 12766442 TI - The Effects of Modeling and Experience on Young Children's Persistence at a Gambling Game. AB - Gambling is common among children and adolescents, but Hide is known about factors initiating or maintaining this behavior. Fifty-one male and 51 female kindergarten and first grade children were invited to play a game involving repeated opportunities to select colored chips from a cup while blindfolded. Children playing for tangible incentives elected to play longer than those who were not (p <.001). Seeing a videotaped model win or fail to win a large prize had no effect on persistence with the game. Playing again one week later, children playing for incentives exhibited a more successful strategy, quitting sooner (p <.04) and with more winnings (p <.03). The parameters of experiential versus observational learning are discussed, with implications for educating children about risk-taking. PMID- 12766445 TI - First Year Impacts of Casino Gambling in a Community. AB - Gambling has become both a major pastime for Canadians and a multibillion dollar industry providing provincial governments with an increasing proportion of their annual revenues. The continuing trend toward the legalization of gambling has made research on the public health impacts of gambling increasingly important to citizens and decision-makers. This article presents first year results of a multi year project to measure the impact of the opening of Casino Windsor on gambling behaviour in Windsor, Ontario, Canada. A random telephone survey of gambling behaviour was conducted with 2,682 adult residents of metropolitan Windsor prior to the opening of Casino Windsor, and was repeated with 2,581 residents one year later. There were no statistically significant changes in the rates of problem and pathological gambling among men, women, or the general population one year following the opening of the casino. Although there was some evidence of higher spending gamblers within the post-casino sample, no statistically significant differences were found between pre- and post-casino per capita gambling expenditures. Implications of these results for the future measurement and treatment of problem and pathological gambling are discussed. PMID- 12766444 TI - Adolescent Gambling Behavior: A Prevalence Study and Examination of the Correlates Associated with Problem Gambling. AB - Eight-hundred and seventeen adolescent high school students in the Montreal region completed the DSM-IV-J gambling screen along with a questionnaire devised by the authors inquiring about their gambling behavior, including items assessing the types of activities in which they engage, frequency of involvement, reasons for gambling, and their cognitive perceptions of gambling activities. The results indicate that, in general, 80.2% of students reported having gambled during the previous year, with 35.1% gambling a minimum of once per week. Adolescents reported participating in gambling behavior more often than any other addictive behavior, including cigarette smoking, alcohol consumption, and illicit drug use. The mean age of onset of gambling behavior for the sample was 11.5 years. The rate of pathological gambling was 4.7% as measured by the DSM-IV-J. Pathological gamblers were more likely to have parents with gambling problems and to be engaging in illegal activities than non-pathological gamblers. Gender differences were evident, with males engaging in gambling activities more than females. Differences in game preferences were found, with males more attracted to sports lottery tickets and sports pool betting and females more attracted to lottery tickets and bingo. Gambling awareness and prevention issues are addressed. PMID- 12766447 TI - Sexual Harassment in Casinos: Effects on Employee Attitudes and Behaviors. AB - This study focuses on sexual harassment and job satisfaction, organizational commitment, and employee turnover among casino employees. It is the first study investigating sexual harassment in the gaming industry. Based on sex-role spillover theory it was expected that sexual harassment has less of an impact on casino employees than on employees in other industries. Six Reno, Nevada casinos participated in the study and 330 responses were generated from casino employees. The study results show that sexual harassment of and by casino employees is perceived to occur at about the same rate as in other industries. Sexually harassed employees were compared to employees who indicated that they had not been sexually harassed. Sexually harassed employees were less satisfied with their jobs and less committed to the organization. However, they were not more likely to quit their jobs. Sexually harassed employees tended to be younger, Caucasian, and in dealer positions. Hence, in addition to the well-publicized cost of sexual harassment lawsuits, the study shows that sexual harassment in casinos may well be the source of hidden costs important to human resources managers. A result of major interest was that employees who had been harassed held management responsible for not ensuring a work environment that is free of sexual harassment. Implications for casino management are discussed. PMID- 12766446 TI - Problem Gambling Within a Chinese Speaking Community. AB - The purpose of this study was to obtain preliminary data on pathological gambling rates within a metropolitan Chinese community to determine if more costly extensive epidemiological surveys were justified. Two thousand survey questionnaires were distributed to parents through children attending a local Chinese speaking school. A response rate of 27.4% was obtained. Over a quarter of respondents were born overseas in either Mainland China or Hong Kong. Results indicated that gambling was not a popular activity with 60.0% of respondents stating that they never gambled. Of those who gambled, a third of respondents identified lotto as the preferred form. Using a Chinese translation version of the SOGS and a cut-off score of 10, a prevalence estimate of 2.9% for pathological gambling was found with males showing a higher rate (4.3%) as compared to females (1.6%). Respondents reporting a prior history of gambling in their country of origin were more likely to be classified as probable pathological gamblers. Despite the use of a sample of convenience, changes to the wording of two items in the Chinese translation of the SOGS and the possibility of false positive cases in the present study, it is concluded that further research into problem gambling in this community should be undertaken. PMID- 12766449 TI - Doubling vs. Constant Bets as Strategies for Gambling. AB - Some gamblers use a doubling strategy as a way of improving their chances of coming home a winner. This paper reports on the results of a computer simulation study of the doubling strategy and compares the short term and long term results of doubling to gambling with a constant sized bet. In the short term players using a doubling strategy were more likely to win, then lose, however in the long term, the losses suffered by doublers were much greater than that suffered by constant bettors. It is argued that the use of a doubling strategy is related to an incomplete conceptualization of random events sometimes known as the 'law of averages.' A second simulation examined the fate of doubling in an ideal world in which the 'law of averages' was actually true. In this ideal world, doublers were much better off than constant bettors. The relationship of the results to a naive conceptualization of random events is discussed. PMID- 12766448 TI - A Case Study on the Informational Efficiency of Markets: The Market for Horse Racing in Australia. AB - This paper describes a study of the informational efficiency of the thoroughbred horse racing market in Australia. It is based on the theory of stock market efficiency which explains the process by which information becomes reflected in share prices. In this paper, the theory is applied to the thoroughbred horse racing market to determine the predictive accuracy of alternative informative sources. The results obtained from the study are consistent with the underlying theory:(i) aggregated information (as reflected in a consensus of opinions) is a more accurate prediction of success than less information (as reflected in individual opinions), and;(ii) the most recent information (as reflected in race time betting odds, known as starting prices) has greater predictive ability than less recent information (as reflected in an earlier consensus of opinions).The study examines predictive accuracy in a gambling context, but does not consider the profitability of alternative prediction processes. PMID- 12766450 TI - Student-Athletes and Gambling: An Analysis of Attitudes Towards Risk-Taking. AB - Gambling scandals at Arizona State University, Boston College, and Northwestern University have made gambling prevention a point of emphasis in the NCAA and throughout colleges and universities across the nation. Despite this emphasis, there is minimal research at any level regarding gambling and student-athletes. This research examines attitudinal differences towards risk-taking among student athletes who gamble on college sports and those who indicate no such gambling activity. Our findings indicate that student-athletes who gambled were more likely to have attitudes supportive of risk taking behavior than their student athlete peers who did not gamble. PMID- 12766451 TI - Alcohol Consumption and Self-Control of Gambling Behaviour. AB - This paper reports the results of a preliminary investigation into whether the drinking of alcohol contributes to impaired control of gambling behaviour. The sampling method consisted of a two-phase survey design, collecting data both prospectively 'within session' and retrospectively via a take-home questionnaire. One hundred sixteen people were interviewed while in a gaming venue playing on an electronic gaming machine (egm) of whom 34 men and 11 women also returned take home surveys. Comparisons of the sample to previous studies suggested that the sample was representative of the population of egm players. Results indicated a consistent theme of alcohol use contributing significantly to impaired control of gambling behaviour, with level of involvement (Corless & Dickerson, 1989) contributing the most significant variance in the independent variables. Limitations of the study are discussed, but the case is argued that this type of 'process' research is essential in better understanding how these two types of popular leisure activities may interact, possibly leading to the previously recorded chronic, excessive alcohol intake and problematic gambling (e.g. McCormick, Russo, Ramirez & Taber, 1984). PMID- 12766453 TI - Criminal Behavior Associated with Pathological Gambling. AB - The influence of addictive gambling behavior on criminal behavior was examined in this study. A sample of pathological gamblers from in- and outpatient treatment centers and self-help groups (n = 300) and a sample of high and low frequency gamblers from the general population and army (n = 274) completed a comprehensive questionnaire which assessed social attachment, personality, pathological gambling and criminal behavior variables. The causal analysis of a Lisrel Model leads to the following results: addictive gambling behavior is an important criminogenic factor. This predisposing factor alone cannot sufficiently explain criminal behavior associated with pathological gambling. Personality variables also directly influence the intensity of criminal behavior. Social attachment variables have only an indirect effect. As far as property offenses are concerned, it was found that the direct causal effect of addiction behavior is greater than that of personality. PMID- 12766452 TI - Narrowing of Attention and Dissociation in Pathological Video Lottery Gamblers. AB - The phenomena of attention and dissociation were investigated in pathological and occasional video lottery (VLT) gamblers. Twelve problem VLT gamblers [mean age 41.3; 6 male, 6 female; South Oaks Gambling Screen (SOGS; Lesieur & Blume, 1987) scores >/= 5] were compared to a group of 11 occasional VLT gamblers, (mean age 31.9; 4 male, 7 female; SOGS scores < 5). Pathological gamblers were slower than occasional gamblers in reacting to irrelevant external light stimuli while playing on a demonstration video lottery terminal. They were significantly more likely to report more symptoms of general dissociation as measured by the Dissociative Experiences Scale (Bernstein & Putnam, 1986). The results suggest that pathological gamblers may experience a greater narrowing of attention than occasional gamblers when engaged in VLT play. PMID- 12766454 TI - Checkered Expectations: Predictors of Approval of Opening a Casino in the Niagara Community. AB - This paper reports on the findings of a survey prior to the opening of a casino in Niagara Falls, Ontario (N= 1002 adults) on approval of the casino, expectations regarding the impact of the casino, attitudes toward gambling, gambling behaviour, and demographic information. The respondents generally had a positive attitude towards gambling. The expectations of community impact clustered into 3 factors: negative social consequences (crimes, addiction), negative environmental consequences (litter, noise, traffic), and positive economic consequences (jobs, stores, income). The majority of respondents expected economic benefits from the casino as well as a decrease in the environmental quality of the city. Expectations regarding social problems were mixed with a majority expecting an increase in serious crimes, but only a minority expecting an increase in people on welfare. Covariance structure modelling revealed that a positive attitude towards gambling and expecting economic benefits were positively related to approval of the casino, and expecting social problems was negatively related to approval. Given that more than seven in ten respondents supported the opening of the casino, the expected economic benefits coupled with a generally positive attitude towards gambling, apparently outweighed concerns about problems associated with gambling. PMID- 12766455 TI - A Comparison of Hispanic and Anglo Calls to a Gambling Help Hotline. AB - Prevalence studies have found that problem gamblers tend to be non-Caucasians. Nevertheless, information on non-Caucasian gambling patterns and problems is virtually non-existent. Data collected during years 1992-1998 on Hispanic (N = 209) and Anglo (N = 5311) problem gamblers calling the Florida Council on Compulsive Gambling Hotline for help is examined to provide information on one such non-Caucasian group: Hispanics. A sharp difference in the number of Hispanic and Anglo callers was found during this six year period (3.8% Hispanic and 96.2% Anglo). Differences were also found in the likelihood of Hispanics calling about their own problems, having gone for previous help, and types of gambling activities. Similarities were found between the two groups regarding age, marital status, and the three most cited problems caused by gambling: problems with family, inability to pay bills and going into debt. Anglos were significantly more likely to "engage in illegal activities for gambling money" and "problems with job." Group differences caution against using Anglo based prevention and treatment programs with Hispanics populations. PMID- 12766456 TI - The UK National Telephone Gambling Helpline-Results on the First Year of Operation. AB - This study outlines the results of the UK's national gambling helpline run by GamCare. The results outlined here cover the period of the first 12 months of operation (November 1997 to October 1998). The helpline received a total of 1729 calls. Of these, 51% were from problem gamblers themselves (90% male; 10% female) and a further 26% of calls were from relatives of problem gamblers. The remaining calls came from other professionals handling problem gambling cases (13%), attempted calls, e.g., people calling and then putting the phone down due to being scared of talking (4%), information requests (3%) and the media (3%). Fruit machine gambling appeared to be most problematic for the callers as a whole and for particular sub-groups such as adolescents (82%) and women (52%). PMID- 12766458 TI - A Comment on Walker and Barnett's "The Social Costs of Gambling: An Economic Perspective" PMID- 12766457 TI - The Social Costs of Gambling: An Economic Perspective. AB - Much of the opposition to legalized gambling is based on analyses of the social costs that occur as a result of pathological gambling. It is our contention that many, if not most, authors who have contributed to this literature are either unclear or misguided in what they define as social costs. Instead of starting with a clear definition of what constitutes a social cost, these authors have adopted an ad hoc approach-using "common sense" to determine what constitutes losses to society and then attempting to quantify the impact of those activities. We believe this is not, as some suggest, simply a matter of semantics. Rather, it is a serious problem in the gambling literature. How do we differentiate between a consequence of pathological gambling that is a "social cost" and one that is not? Which of the consequences of addictive behaviors that are associated with gambling arise when gambling is legal, and which will be manifest in some form whether or not gambling is legal? In this article we explain the economic perspective on social costs. An understanding of this paradigm removes the subjectivity in the classification of pathological gambling's social costs. The paper has three major components. First, we introduce the economic notion of social costs. Using this paradigm, we differentiate between the "true" social costs related to pathological gambling, and other negative consequences that cannot legitimately be classified as social costs. Second, we evaluate a recent social cost study using the economics social cost paradigm. Third, we discuss two types of social costs that have been largely overlooked in the gambling literature. One is caused by gambling prohibition. The other occurs as a result of "rent seeking" that is related to the political process surrounding the legalization of gambling. PMID- 12766459 TI - Response to McGowan's Comment on "The Social Costs of Gambling: An Economic Perspective" PMID- 12766460 TI - An Epidemiologic Note on Verification Bias: Implications for Estimation of Rates. AB - Two-stage designs offer several advantages for purposes of test validation and prevalence estimation. These include enhanced precision and increased cost efficiency. Cost efficiency is obtained when the best available verification criterion is too expensive to employ in a large-scale epidemiologic study. The use of two-stage (or double-sampling) designs permit the same inferences from application to only a subset of those who were screened (with the less expensive criterion) during stage one. The retesting of only some of the first stage respondents introduces a bias, however, if these are sampled on the basis of first stage screening results. The form of this bias is described and solutions for correcting estimates are provided. These solutions are applied to the data reported by Abbott and Volberg (1996) in their study of the New Zealand general population. Corrected estimates of the sensitivity and specificity of the South Oaks Gambling Screen are obtained and used to adjust the reported lifetime and current prevalence estimates. The value of multi-stage designs for validity assessment and prevalence estimation are briefly described. PMID- 12766461 TI - A Reply to Gambino's "An Epidemiologic Note on Verification Bias: Implications for Estimation of Rates" AB - While two-stage designs in problem gambling research are expensive and relatively difficult to execute, they have the potential to yield more accurate prevalence estimates as well as valuable additional information about gambling and problem gambling in the community. However, gains in precision are heavily dependent on the accuracy of the screening methods used and the sample sizes involved. Sound practice requires reliable information about the variances and confidence levels associated with different screens. In the spirit of contributing to the ongoing dialog about ways to improve the measurement of problem gambling, we examine several reasons to question whether the revised estimates offered by Gambino are in fact an improvement. PMID- 12766462 TI - Estimating Confidence Intervals and Sampling Proportions in Two-Stage Prevalence Designs. AB - In response to Abbott and Volberg's (in press) rejoinder to my epidemiologic note on verification bias and estimation of prevalence rates (Gambino, in press), I provide the formulas for computing confidence intervals for the results of second stage verification. In addition, I provide the appropriate equation for determining confidence intervals when prevalence is near zero or one. Finally, we present formulas for determining the most efficient sample sizes needed to minimize second-stage variance estimates. These allow the investigator working under a fixed budget to determine the relative value of sampling negative screens to test for false negatives. We close with an observation on the interpretability of evidence. PMID- 12766463 TI - A Case Study of a Casino Campaign: Testing the Dombrink-Thompson Model. PMID- 12766464 TI - Gambling Technologies: Prospects for Problem Gambling. AB - Technology his always played a role in the development of gambling practices and will continue to play a critical role in the development of increased gambling opportunities (e.g., internet gambling). Although technological advance his long been associated with improved gambling opportunities, there is little written in the literature explicitly pointing out this link and its implications for problem gamblers. This paper therefore reviews this situation and examines the technological implications of situational and structural characteristics paying particular attention to slot machine gambling as there has been more empirical work on this type of gambling than any other technological form. The impact of technology on the sociability of gambling is also examined followed by a more speculative evolution of internet gambling as an area of potential concern. PMID- 12766465 TI - Gambling: Achieving the Right Balance. PMID- 12766466 TI - Gambling and the Health of the Public: Adopting a Public Health Perspective. AB - During the last decade there has been an unprecedented expansion of legalized gambling throughout North America. Three primary forces appear to be motivating this growth: (1) the desire of governments to identify new sources of revenue without invoking new or higher taxes; (2) tourism entrepreneurs developing new destinations for entertainment and leisure; and (3) the rise of new technologies and forms of gambling (e.g., video lottery terminals, powerball mega-lotteries, and computer offshore gambling). Associated with this phenomenon, there has been an increase in the prevalence of problem and pathological gambling among the general adult population, as well as a sustained high level of gambling-related problems among youth. To date there has been little dialogue within the public health sector in particular, or among health care practitioners in general, about the potential health impact of gambling or gambling-related problems. This article encourages the adoption of a public health perspective towards gambling. More specifically, this discussion has four primary objectives:1. Create awareness among health professionals about gambling, its rapid expansion and its relationship with the health care system;2. Place gambling within a public health framework by examining it from several perspectives, including population health, human ecology and addictive behaviors;3. Outline the major public health issues about how gambling can affect individuals, families and communities;4. Propose an agenda for strengthening policy, prevention and treatment practices through greater public health involvement, using the framework of The Ottawa Charter for Health Promotion as a guide.By understanding gambling and its potential impacts on the public's health, policy makers and health practitioners can minimize gambling's negative impacts and appreciate its potential benefits. PMID- 12766467 TI - The social costs and benefits of gambling: an introduction to the economic issues. AB - This paper surveys the range of economic issues that need to be addressed in attempting to evaluate the social costs and benefits of gambling. It considers, inter alia, the nature of private and social costs and benefits and their policy significance, the important distinction between real and pecuniary costs, the incidence of gambling costs and benefits and the sources of gambling costs and benefits. Similarities and differences between the analysis of gambling and alcohol are considered. The paper concludes with a consideration of why various estimates of the social costs of gambling have produced such divergent results. PMID- 12766468 TI - Methodological issues in the social cost of gambling studies. AB - The appropriate way to classify and measure the "social costs" of gambling is a very important, unresolved methodological issue that has been addressed by Collins and Lapsley (2000); Thompson, Gazel, and Rickman (1999); and Walker and Barnett (1999), among others. What should be included and excluded from social cost studies continues to be a controversial issue, as illustrated in the literature and recent conferences. This paper is an attempt to explain the "economics" conception of social costs in accessible language. By using a simple economic model and everyday examples, it shows that the economics methodology is better than the other methodologies currently available. There are four specific goals of the paper: (1) Discuss the importance of the social cost methodological debate and the state of research in the area; (2) Explain the Walker-Barnett definition of social cost in the context of a simple production possibilities frontier and indifference curve model; (3) Use simple illustrative examples to show why many of the alleged social costs should not be classified as such; and (4) Suggest a new method for analyzing the social costs and effects attributable to pathological gambling. PMID- 12766469 TI - Measuring costs from permitted gaming: concepts and categories in evaluating gambling's consequences. AB - This paper addresses the methodology of cost-benefit analysis as applied to policy alternatives regarding legally sanctioned gambling in its various forms. Existing economic studies regarding the social costs of gambling are reviewed and critiqued. Distinctions are made between definitions of social costs that are defined as actions which result in negative changes in aggregate social wealth (the "narrow" definition), and those which also include internal nonmarket costs that are borne by individual gamblers and their immediate families and acquaintances (the "broader" definition). This distinction is important because of its bearing on economic policies that are primarily concerned with economic efficiency versus policies that are more paternalistic, which attempt to protect individuals from self-damage or self-destruction by restricting their ranges of choice. Whether societies choose to prohibit or severely restrict permitted gambling, or allocate substantial resources to mitigate its negative side effects, rests largely on which of these perspectives regarding social costs they find more appropriate. Finally, the issue of social protection through restrictions on the availability of gambling to the entire population, versus a strategy tailored toward identifiable "problem" gamblers, is discussed. PMID- 12766470 TI - Estimating the costs of substance abuse: implications to the estimation of the costs and benefits of gambling. AB - This presentation describes a recently developed set of guidelines for estimating the economic costs of substance abuse, summarizes the findings from a Canadian study that utilized these guidelines, and discusses the implications to the potential development of guidelines for estimating the costs and benefits of gambling. The guidelines for estimating the costs of substance abuse present a general framework of costs to be included and discuss methodological issues such as the definition of abuse; determination of causality; comparison of the demographic and human capital approaches; the treatment of private costs; the treatment of nonworkforce mortality and morbidity; the treatment of research, education, law enforcement costs, the estimation of avoidable costs and budgetary impact of substance abuse, and the significance of intangible costs. Utilizing these guidelines, a study was undertaken to estimate the economic costs of alcohol, tobacco, and illicit drugs to Canadian society in 1992. Based on this experience, it is argued that cost/benefit research should be conducted by multidisciplinary teams, that the real value of such work lies more in the detailed findings than in the bottom line results, and that focus should be placed on developing an ongoing process for developing consensus on how to conduct studies of the costs and benefits of gambling, rather than attempting to find a precise methodology that everyone agrees upon. PMID- 12766471 TI - Framing public policy towards a public health paradigm for gambling. AB - This paper examines the public policy value of looking at gambling from a public health perspective. The manner in which social issues are framed will either expand or curtail public policy debates. The existing and traditional frames for gambling (e.g. gambling as a matter of individual freedom, gambling as a form of recreation) fail to consider research on the social and economic impacts of gambling. Because a public health frame offers a broad viewpoint of society, it encompasses a number of social and economic impacts not considered in traditional frames. However, the existing gambling frames enjoy varying degrees of cultural, economic, and political support and, as a result, creating a higher profile for a public health framework will encounter a number of barriers. Research can play a decisive role in overcoming these barriers, as it has in a number of related fields (e.g., tobacco use, addiction and product liability, the epidemiology of AIDS). The paper concludes that research that identifies and quantifies the public health factors of gambling will substantially contribute to a public shift toward a public health frame. PMID- 12766472 TI - The mitochondrial membrane potential (deltapsi(m)) in apoptosis; an update. AB - Mitochondrial dysfunction has been shown to participate in the induction of apoptosis and has even been suggested to be central to the apoptotic pathway. Indeed, opening of the mitochondrial permeability transition pore has been demonstrated to induce depolarization of the transmembrane potential (deltapsi(m)), release of apoptogenic factors and loss of oxidative phosphorylation. In some apoptotic systems, loss of deltapsi(m) may be an early event in the apoptotic process. However, there are emerging data suggesting that, depending on the model of apoptosis, the loss of deltapsi(m) may not be an early requirement for apoptosis, but on the contrary may be a consequence of the apoptotic-signaling pathway. Furthermore, to add to these conflicting data, loss of deltapsi(m) has been demonstrated to not be required for cytochrome c release, whereas release of apoptosis inducing factor AIF is dependent upon disruption of deltapsi(m) early in the apoptotic pathway. Together, the existing literature suggests that depending on the cell system under investigation and the apoptotic stimuli used, dissipation of deltapsi(m) may or may not be an early event in the apoptotic pathway. Discrepancies in this area of apoptosis research may be attributed to the fluorochromes used to detect deltapsi(m). Differential degrees of sensitivity of these fluorochromes exist, and there are also important factors that contribute to their ability to accurately discriminate changes in deltapsi(m). PMID- 12766473 TI - Drosophila morgue and the intersection between protein ubiquitination and programmed cell death. AB - In Drosophila, cell survival decisions are mediated by the integrated functions of the Grim-Reaper death activators and Inhibitor-of-Apoptosis-Proteins (IAPs), such as DIAP1, to regulate caspase activities. We recently identified a gene that enhances the actions of the Grim-Reaper proteins and negatively regulates the levels of DIAP1 protein. This gene, morgue, encodes a novel protein that contains both an F box and a ubiquitin conjugase domain. Interestingly, the Morgue conjugase domain lacks the active site cysteine required for covalent linkage to ubiquitin. Morgue could target IAPs and other proteins for ubiquitination and proteasome-dependent turnover by acting either in an SCF ubiquitin E3 ligase complex, or as a ubiquitin E2 conjugase enzyme variant (UEV) in conjunction with a catalytically active E2 conjugase. Morgue is evolutionarily conserved, as a Morgue ortholog was identified from the mosquito, Anopheles gambiae. Elucidation of morgue function should provide novel insights into the mechanisms of ubiquitination and programmed cell death. PMID- 12766475 TI - Lack of FasL-mediated killing leads to in vivo tumor promotion in mouse Lewis lung cancer. AB - Lewis lung carcinoma (3LL) cells were constitutively resistant to Fas-mediated apoptosis, but overexpression of Fas on 3LL cells allowed Fas-mediated apoptosis after crosslinking with agonist anti-Fas antibody (Jo2) in vitro. Surprisingly, Fas-overexpressing 3LL cells showed enhanced in vivo tumor progression, whereas no promotion of in vivo tumor growth was observed for dominant negative (DN) Fas overexpressing 3LL transfectants in which the cytoplasmic death domain was deleted. In addition, the promotion of in vivo tumor growth by Fas-overexpression was reduced in gld (FasL-mutation) mice compared to normal mice. These data indicate that intact Fas/FasL cell signaling is required for the promotion of in vivo tumor growth by Fas overexpression in 3LL cells. In contrast to the efficient Fas-mediated killing induced in vitro by crosslinking with anti-Fas antibody, Fas-overexpressing 3LL cells were resistant in vitro to Fas-mediated apoptosis by activated T cells or transient FasL transfection. These data suggest that agonist anti-Fas antibody and natural FasL can transmit qualitatively different signals, and crosslinking of Fas with natural FasL on 3LL cells does not deliver the expected death signal. Thus, our results demonstrate that in some cases overexpression of Fas can result in a survival advantage for tumor cells in vivo. PMID- 12766474 TI - Reovirus-induced apoptosis: A minireview. AB - Reoviruses infect a variety of mammalian hosts and serve as an important experimental system for studying the mechanisms of virus-induced injury. Reovirus infection induces apoptosis in cultured cells in vitro and in target tissues in vivo, including the heart and central nervous system (CNS). In epithelial cells, reovirus-induced apoptosis involves the release of tumor necrosis factor (TNF) related apoptosis-inducing ligand (TRAIL) from infected cells and the activation of TRAIL-associated death receptors (DRs) DR4 and DR5. DR activation is followed by activation of caspase 8, cleavage of Bid, and the subsequent release of pro apoptotic mitochondrial factors. By contrast, in neurons, reovirus-induced apoptosis involves a wider array of DRs, including TNFR and Fas, and the mitochondria appear to play a less critical role. These results show that reoviruses induce apoptotic pathways in a cell and tissue specific manner. In vivo there is an excellent correlation between the location of viral infection, the presence of tissue injury and apoptosis, indicating that apoptosis is a critical mechanism by which disease is triggered in the host. These studies suggest that inhibition of apoptosis may provide a novel strategy for limiting virus-induced tissue damage following infection. PMID- 12766476 TI - Induction of apoptosis by a cachectic-factor in murine myotubes and inhibition by eicosapentaenoic acid. AB - Treatment of C(2)C(12) myotubes with a tumour-derived proteolysis-inducing factor (PIF) at concentrations between 1 and 10 nM was shown to stimulate the activity of the apoptotic initiator caspases-8 and -9 and the apoptotic effector caspases 2, -3 and -6. This increased caspase activity was attenuated in myotubes pretreated with 50 microM eicosapentaenoic acid (EPA). At least part of the increase in caspase activity may be related to the increased proteasome proteolytic activity, since a caspase-3 inhibitor completely attenuated the PIF induced increase in 'chymotrypsin-like' enzyme activity, the predominant proteolytic activity of the proteasome. However, Western blot analysis showed that PIF induced an increase in expression of the active form of caspase-3, which was also attenuated by EPA. Further Western blot analysis showed PIF increased the cytosolic content of cytochrome c, as well as expression of the pro-apoptotic protein bax but not the anti-apoptotic protein bcl-2, which were both attenuated by 50 microM EPA. Induction of apoptosis by PIF in murine myotubes was confirmed by an increase in free nucleasomes formation and increased DNA fragmentation evidenced by a nucleasomal ladder typical of apoptotic cells. This process was again inhibited by pre-incubation with EPA. These results suggest that in addition to activating the proteasome, PIF induces apoptosis in C(2)C(12) myotubes, possibly through the common intermediate arachidonic acid. Both of these processes would contribute to the loss of skeletal muscle in cancer cachexia. PMID- 12766477 TI - Glucocorticoid-induced apoptosis in lymphoid organs is associated with a delayed increase in circulating deoxyribonucleic acid. AB - Pathological processes like cancer, chronic inflammation and autoimmune phenomena, all of which involve massive cell death, are associated with significant increases in circulating DNA. In order to clarify whether massive apoptosis occurring under physiological circumstances also causes DNA release into the circulation, we correlated the time-course of dexamethasone-induced intra thymic cell apoptosis with plasma DNA dynamics in rats. Animals were given 10 mg/l dexamethasone in their drinking water for up to 7 days. Sequential plasma samples were obtained during the treatment and DNA was quantitated by a micro fluorometric assay. Thymus and spleen weight as well as apoptotic cell levels were assessed at different times. Seven days of glucocorticoid treatment reduced thymic and spleen mass by 82 and 31%, respectively. Intra thymic apoptosis was maximal 24 h after the beginning of glucocorticoid treatment, declining markedly by 48 h. Very little apoptosis was observed in the spleen. Plasma DNA increased steadily during the first 4 days of glucocorticoid treatment (11.8 +/- 1.2 microg/ml on day 0; 24.2 +/- 1.6 microg/ml on day 4) beginning to decline afterward. Thymectomy but not splenectomy, drastically reduced the glucocorticoid induced increase in plasma DNA. It is concluded that hormone-induced massive intra thymic cell death is followed by a delayed release of nucleosomal DNA into the circulation. PMID- 12766478 TI - Potential role of the EPEC translocated intimin receptor (Tir) in host apoptotic events. AB - Apoptosis, or programmed cell death, is a well-ordered process that allows damaged or diseased cells to be removed from an organism without severe inflammatory reactions. Multiple factors, including microbial infection, can induce programmed death and trigger reactions in both host and microbial cellular pathways. Whereas an ultimate outcome is host cell death, these apoptotic triggering mechanisms may also facilitate microbial spread and prolong infection. To gain a better understanding of the complex events of host cell response to microbial infection, we investigated the molecular role of the microorganism Enteropathogenic Escherichia coli (EPEC) in programmed cell death. We report that wild type strain of EPEC, E2348/69, induced apoptosis in cultured PtK2 and Caco-2 cells, and in contrast, infections by the intracellularly localized Listeria monocytogenes did not. Fractionation and concentration of EPEC-secreted proteins demonstrated that soluble protein factors expressed by the bacteria were capable of inducing the apoptotic events in the absence of organism attachment, suggesting adherence is not required to induce host cell death. Among the known EPEC proteins secreted via the Type III secretion (TTS) system, we identified the translocated intimin receptor (Tir) in the apoptosis-inducing protein sample. In addition, host cell ectopic expression of an EPEC GFP-Tir showed mitochondrial localization of the protein and produced apoptotic effects in transfected cells. Taken together, these results suggest a potential EPEC Tir-mediated role in the apoptotic signaling cascade of infected host cells. PMID- 12766479 TI - Apoptosis staining in cultured pseudoachondroplasia chondrocytes. AB - Pseudoachondroplasia (PSACH) is a skeletal dysplasia caused by a mutation in cartilage oligomeric matrix protein (COMP), a glycoprotein of normal cartilage matrix. PSACH chondrocytes have a distinctive phenotype with enlarged rER cisternae containing COMP, aggrecan, type IX collagen, and chaperone proteins. Ultrastructural studies suggested that this accumulation compromises cell function, hastening cell death, and consequently reducing the number of cells in the growth plate contributing to linear bone growth. Using the alginate bead system, we cultured control and PSACH chondrocytes for twenty weeks and one year to determine the effect of the mutation on size and number of cartilage nodules; and the presence of apoptotic cell death (TUNEL assay). At 20 weeks, beads containing PSACH or control chondrocytes did not differ in size and number of cartilage nodules or number of TUNEL-positive cells. After one year, nodule number, size and percent cartilage per bead were significantly less in PSACH nodules, and the number of cells staining positive for apoptosis was significantly greater than in controls (71.8% vs. 44.6%). The increase in apoptosis in PSACH nodules correlates with a decrease in growth of cartilage, supporting our hypothesis that death of damaged cells contributes to the growth plate defects in PSACH. PMID- 12766480 TI - [3H]-thymidine labelling of DNA triggers apoptosis potentiated by E1A-adenoviral protein. AB - [(3)H]-thymidine is commonly used to analyze the accumulation of [(3)H]-labeled chromatin fragments in cells undergoing apoptosis. This study shows that [(3)H] thymidine incorporation within DNA is sufficient per se to inhibit growth and to induce apoptosis in canine kidney epithelial cells and porcine aorta endothelial cells. Despite high-level [(3)H]-thymidine-DNA labeling, rat vascular smooth muscle cells (VSMC) showed only modest inhibition of growth and induction of apoptosis compared to other cell types. Similarly to serum deprivation, apoptosis triggered by [(3)H]-thymidine labeling was sharply potentiated by VSMC transfection with a functional analogue of c-myc, E1A-adenoviral protein (VSMC E1A), and was suppressed by stimulation of cAMP signaling with forskolin as well as by and Na/K pump inhibition with ouabain. Both apoptosis induction and growth suppression seen in [(3)H]-thymidine-treated VSMC-E1A were reduced by the pan caspase inhibitor z-VAD.fmk. Thus, our results show that the differential efficiency of the apoptotic machinery determines cell type-specific attenuation of growth in cells with [(3)H]-thymidine-labeled DNA. They also demonstrate that [(3)H]-thymidine-treated and serum-deprived VSMC employ common intermediates of the apoptotic machinery, including steps that are potentiated by E1A-adenoviral protein and inhibited by activation of cAMP signaling as well as by inversion of the intracellular [Na(+)](i)/[K(+)](i) ratio. PMID- 12766481 TI - A genetic selection method for expression products that induce apoptosis in adherent mammalian cell lines. AB - The process of apoptosis is carefully controlled in cells, and different cell types display different sensitivities to pro-apoptotic stimuli. The prospect of exploiting such differences for treatment of diseases such as cancer, via novel therapeutic agents, is extremely attractive. Therefore, genetic selections for novel expression products that kill cells may have considerable value. However, such selections are difficult to devise and perform because the selected cells do not grow. We developed a selection scheme designed to enrich for genetic agents that kill cells. The selection is based on detachment of apoptotic cultured mammalian cells from adherent monolayers. We characterized the properties of these detached cells (floating cells), and various aspects of the selection process. This selection method is potentially applicable to many mammalian cell lines. PMID- 12766482 TI - The role of ATF/CREB family members in cell growth, survival and apoptosis. AB - Programmed cell death is an integral part of the mechanisms regulating tissue homeostasis. Defects in the apoptotic signaling pathway are often associated with uncontrolled cell proliferation, high mutation rate and malignant transformation. Transcription factors, such as the mammalian ATF/CREB family of transcriptional regulators, have diverse functions in controlling cell proliferation and apoptosis. One particular ATF/CREB family member, ATFx, is an anti-apoptotic factor that plays an essential role in cell survival. Current observations indicate that one mechanism by which ATFx inhibits cell death and promotes cell survival is by disrupting signal transmission from activated "death receptors" to initiator caspases. A better understanding of ATFx function should provide new insight into the processes that control apoptotic cascades. PMID- 12766483 TI - BNips: a group of pro-apoptotic proteins in the Bcl-2 family. AB - BNip (formerly known as Nip) proteins, including homologues isolated from human, mouse and Caenorhabditis. elegans, are a relatively new subgroup of the Bcl-2 family. These proteins are classified into this family based on limited sequence homology with the Bcl-2 homology domain 3 and carboxyl terminal transmembrane domain. BNip proteins were first discovered based on their interaction with the adenovirus E1B 19 kDa/Bcl-2 family protein and since then, their roles in cell death pathways have been actively studied. However, the precise mechanisms by which the BNip proteins induce apoptosis and/or necrosis remain to be determined. To advance our knowledge, we have provided a summary and review of current literature regarding BNip proteins including comparative sequence analysis, mutational mapping of the functional domains, and cell death mechanisms involving disruption of mitochondrial homeostasis. Since BNip proteins are expressed at high levels in the heart as compared to other organs, their roles in cardiomyocyte injury during hypoxia or viral infection is a focus of this review. Finally, we discuss potential directions for further study on this increasingly important group of pro-apoptotic proteins. PMID- 12766484 TI - Apoptosis and interferons: role of interferon-stimulated genes as mediators of apoptosis. AB - IFNs are a family of cytokines with pleiotropic biological effects mediated by scores of responsive genes. IFNs were the first human proteins to be effective in cancer therapy and were among the first recombinant DNA products to be used clinically. Both quality and quantity of life has been improved in response to IFNs in various malignancies. Despite its beneficial effects, unraveling the mechanisms of the anti-tumor effects of IFN has proven to be a complex task. IFNs may mediate anti-tumor effects either indirectly by modulating immunomodulatory and anti-angiogenic responses or by directly affecting proliferation or cellular differentiation of tumor cells. Both direct or indirect effects of IFNs result from induction of a subset of genes, called IFN stimulated genes (ISGs). In addition to the ISGs implicated in anti-viral, anti-angiogenic, immunomodulatory and cell cycle inhibitory effects, oligonucleotide microarray studies have identified ISGs with apoptotic functions. These include TNF-alpha related apoptosis inducing ligand (TRAIL/Apo2L), Fas/FasL, XIAP associated factor-1 (XAF 1), caspase-4, caspase-8, dsRNA activated protein kinase (PKR), 2'5'A oligoadenylate synthetase (OAS), death activating protein kinases (DAP kinase), phospholipid scramblase, galectin 9, IFN regulatory factors (IRFs), promyelocytic leukemia gene (PML) and regulators of IFN induced death (RIDs). In vitro IFN alpha, IFN-beta and IFN-gamma induced apoptosis in multiple cell lines of varied histologies. This review will emphasize possible mechanisms and the role of ISGs involved in mediating apoptotic function of IFNs. PMID- 12766485 TI - Mechanisms of vanilloid-induced apoptosis. AB - Chemical compounds that contain the vanillyl moiety (4-hydroxy-3-methoxybenzyl) are collectively classified as vanilloids. Vanilloid phytochemicals can be found in a variety of sources, some of which are routinely consumed by humans throughout the world. The dietary and/or medicinal use of vanilloids may be effective in inhibiting or reversing carcinogenesis, which has sparked a considerable interest in these compounds as potential chemopreventive or chemotherapeutic agents. Certain vanilloids are also valuable as pharmacological tools for investigating neurobiology, and have been proven effective in alleviating neurogenic pain and inflammation. Recently several vanilloids have demonstrated the ability to induce apoptosis in various cell types. Vanilloids can interact with proteins and membranes to initiate pleiotropic effects, some of which are potentially cytotoxic. Certain vanilloids bind to cation channels on nociceptive sensory neurons to regulate Ca(2+) uptake, which can promote neurotoxicity resulting in apoptosis and necrosis. Furthermore, some vanilloids appear to interfere with enzymatic processes in the plasma membrane and the mitochondria by functioning as coenzyme Q antagonist. This can promote reactive oxygen species production and/or the disruption of redox homeostasis resulting in apoptosis. This review will examine the cellular targets, cytotoxic effects, and the downstream effector mechanisms associated with vanilloid-induced apoptosis. PMID- 12766486 TI - A novel assay for discovery and characterization of pro-apoptotic drugs and for monitoring apoptosis in patient sera. AB - We have developed an apoptosis assay based on measurement of a neoepitope of cytokeratin-18 (CK18-Asp396) exposed after caspase-cleavage and detected by the monoclonal antibody M30. The total amount of caspase-cleaved CK18 which has accumulated in cells and tissue culture media during apoptosis is measured by ELISA. The sensitivity is sufficient for use in the 96-well format to allow high through-put screening of drug libraries. We here describe strategies allowing classification of pro-apoptotic compounds according to their profiles of induction of apoptosis in the presence of pharmacological inhibitors. The time course of induction of CK18 cleavage can furthermore be used to distinguish structurally similar compounds. We propose that compounds that induce rapid CK18 cleavage have mechanisms of actions distinct from conventional genotoxic and microtubuli-targeting agents, and we present one example of an agent that induces almost immediate mitochondrial depolarization and cytochrome c release. Finally, CK18-Asp396 cleavage products are released from cells in tissue culture, and presumably from tumor cells in vivo. These products can be measured in sera from cancer patients. We present evidence suggesting that it will be possible to use the M30-ELISA assay for measuring chemotherapy-induced apoptosis in patient sera, opening possibilities for monitoring therapy. PMID- 12766488 TI - Helix 6 of tBid is necessary but not sufficient for mitochondrial binding activity. AB - The apoptosis effector Bid regulates cell death at the level of mitochondrial cytochrome c efflux. Bid consists of 8 alpha-helices (designated H1 through H8, respectively) and is a soluble cytosolic protein in its native state. Proteolysis of the N-terminus (encompassing H1 and H2) of Bid yields activated "tBid" (truncated Bid), which translocates to the mitochondria and induces the efflux of cytochrome c. Here, we demonstrate that helix H6 of tBid is necessary, albeit not sufficient, for mitochondrial binding. In particular, a 33 amino acid long domain, which encompassed H6 and H7, behaved as the minimum domain in tBid that was sufficient for mitochondrial binding. Unexpectedly, the hydrophobic surface of these helices could be mutated without altering the binding activity of the domain, implying that the secondary structure of the helices may be the key determinant of binding. These experiments expand our mechanistic understanding of the apoptotic regulator, tBid. PMID- 12766489 TI - Sustained release of Smac/DIABLO from mitochondria commits to undergo UVB-induced apoptosis. AB - Apoptotic response of keratinocytes to UVB irradiation has physiological significance on photocarcinogenesis. Here, we show that the sustained release of Smac/DIABLO from mitochondria is an important event for the onset of apoptosis in keratinocytes exposed to UVB irradiation. In human keratinocyte HaCaT cells, UVB irradiation at 500 J/m(2), but not at 150 J/m(2), induces apoptosis. Significant activations of caspases-9 and -3, and slight activation of caspase-7 were observed only in 500 J/m(2) UVB irradiated HaCaT cells. Correspondingly, the cleavage of PARP, a substrate of caspases-3 and -7, was detected in cells irradiated at 500 J/m(2) UVB, but not at 150 J/m(2). However, with both 150 and 500 J/m(2) UVB irradiation, cytochrome c, an activator of caspase-9 via the formation of apoptosome, was released from mitochondria to the cytosol at the same extent. In contrast, significant amounts of Smac/DIABLO are released from mitochondria to the cytosol only with 500 J/m(2) UVB irradiation, and that the level of XIAP is decreased. These results suggest that the extent of Smac/DIABLO efflux from mitochondria is a determinant whether a cell will undergo apoptosis or survival. PMID- 12766487 TI - Rapid induction of apoptosis in human keratinocytes with the photosensitizer QLT0074 via a direct mitochondrial action. AB - QLT0074 is a newly introduced, porphyrin-derivative for use in photodynamic therapy (PDT). In the current study, the intracellular distribution of QLT0074 and the mode of cell death induced by photosensitization with this compound in vitro were assessed for transformed human HaCaT keratinocytes. Fluorescence microscopy studies indicated a distribution of the drug to the cytoplasm, nuclear membrane and mitochondria of these cells. In the absence of light, QLT0074 produced no evidence of apoptosis-related biochemical changes or affected cell viability. When combined with blue light exposure, cytotoxicity was exerted in a QLT0074- and light-dose-related manner. Appearance of the mitochondrial protein cytochrome c in the cytosolic fraction and expression of the apoptosis-associated mitochondrial 7A6 antigen were demonstrable following photosensitization at nano molar levels of QLT0074. Evidence of processing of the apoptosis-effector molecules caspase-3, -6, -7, -8 and -9 as well as cleavage of the caspase-3 substrate poly (ADP-ribose) polymerase (PARP) were demonstrable subsequent to cytochrome c release after PDT. Treatment with the anti-oxidant pyrrolidine dithiocarbamate (PDTC) inhibited cytochrome c release, caspase-3 activation and PARP cleavage associated with PDT thereby supporting the contention that QLT0074 induces apoptosis through the generation of reactive oxygen species upon light activation. QLT0074 is a potent photosensitizer with the capacity to directly initiate apoptosis by acting upon mitochondria. PMID- 12766490 TI - Lactacystin augments the sulindac-induced apoptosis in HT-29 cells. AB - The present study was conducted to explore the potential role of proteasome pathway in NSAIDs-induced apoptosis. We employed sulindac as a NSAID, and chose the lactacystin for inhibition of proteasome activity. Assessment of apoptosis and proteasome activity assay were undertaken. We demonstrated that sulindac treatment resulted in a decrease of proteasome activity, and that the co treatment of a proteasome inhibitor lactacystin potentiated the extent of sulindac-induced apoptosis in HT-29 cells by augmentation of the decrease in proteasome activity. Elucidation of the mechanism underlying the regression of colon cancers by combinations of sulindac and lactacystin seems to be an immediate challenge for the near future. PMID- 12766491 TI - Mechanisms of cell death and disease: advances in therapeutic intervention. PMID- 12766494 TI - Myocardial stunning in man. AB - Myocardial stunning refers to the contractile dysfunction that occurs following an episode of acute ischaemia, despite the return of normal blood flow. The phenomenon was initially identified in animal models, where it has been very well characterised, and there was initial doubt about whether a similar syndrome occurred in humans, and if it did, whether it was of any clinical relevance. This article outlines the conditions that must be met to diagnose myocardial stunning and why it has been difficult to confirm its presence in humans. The clinical scenarios where it has now been clearly identified and those others where it may also occur and be of clinical importance are also reviewed. PMID- 12766492 TI - Effects of brief ischemia and reperfusion on the myocardium and the role of nitric oxide. AB - Brief myocardial ischemia/reperfusion has complex effects on the myocardium. In the short term the myocardium may be stunned with temporarily reduced contractile function, though this may also be accompanied by the modification and de novo synthesis of proteins that protect the heart against subsequent early or delayed insults. Repeated episodes of non-lethal ischemia, which are common in the clinical setting, combine all of these phenomena and may ultimately result in chronic contractile dysfunction. Nitric oxide is intimately linked to many of these alterations in cellular function and defense. This article examines data predominantly from in vivo large animal studies that relate to these ischemia induced changes, the evidence for the proposed mechanisms behind both myocardial stunning and preconditioning while concentrating on the role of nitric oxide in these conditions. PMID- 12766495 TI - Evidence that stunning can be cumulative in man. AB - Myocardial stunning and hibernation are two entities that have become increasingly recognised as clinically important causes of reversible left ventricular (LV) dysfunction. Their occurrence is important as resting myocardial dysfunction, which was once thought to be irreversible, may recover if ischaemia is lessened or abolished. Recent evidence has suggested that cumulative stunning can occur in man and may in fact be responsible for the phenomenon of hibernation. In this chapter we will review the evidence supporting the occurrence of cumulative stunning in man. PMID- 12766493 TI - Novel mechanisms mediating stunned myocardium. AB - Myocardial stunning is defined as the prolonged contractile dysfunction following an ischemic episode that does not result in necrosis, which also occurs in patients with coronary artery disease. There is also evidence to consider myocardial stunning as a fundamental component of hibernating myocardium. Various experimental approaches (from a brief episode to prolonged partial ischemia) and animal models (from rodents to large mammals) have been developed to investigate the pathogenesis of myocardial stunning. Three hypotheses to explain the mechanism, i.e. oxygen radical, Troponin I degradation, and Ca(2+), have been proposed. The first was tested primarily using large mammalian models, whereas the others were tested primarily using rodent models. Recently, the Ca(2+) handling hyothesis has been tested in a large mammalian swine model of myocardial stunning, in which both Ca(2+) and transients and L-type Ca(2+) current density were decreased. Relaxation function and phospholamban phosphorylation are also radically different in large mammalian and rodent models. In addition, troponin I degradation, which was identified as the mechanism of stunning in rodent models, was not found in stunned swine myocardium. Interestingly, the large mammalian model demonstrates that stunning elicits broad changes in gene and protein regulation, some of which have not been observed in the heart previously. The overall genomic adaptation upregulates the expression of survival genes that prevent irreversible damage. Pursuing these new concepts derived from large mammalian models of ischemia/reperfusion will provide more comprehensive mechanistic information underlying myocardial stunning and will serve to devise new therapeutic modalities for patients. PMID- 12766496 TI - Hibernation and congestive heart failure. AB - The most common cause of heart failure is coronary artery disease, and whilst intensive treatment of acute coronary syndromes and myocardial infarction continue to reduce the mortality associated with these conditions, many survivors develop heart failure. In general, heart failure secondary to ischaemic heart disease results from: (i) irreversible myocyte loss due to infarction with scar formation; (ii) chronic left ventricular dysfunction which may recover after revascularisation (hibernating myocardium); (iii) changes in remote myocardium (adverse remodelling). A number of studies suggest that patients with post ischaemic heart failure may derive symptomatic and prognostic benefit from coronary revascularisation and most of this benefit is thought to derive from functional improvement of hibernating myocardium. Although the mechanisms of hibernation remain poorly understood, studies with positron emission tomography have shown that blood flow to hibernating myocardium is usually within or only slightly below the normal range whilst the coronary vasodilator reserve is always severely reduced and the concept that stunning and hibernation may be causally related has gained support in recent years. There is increasing consensus amongst clinicians regarding the importance of identifying and treating hibernating myocardium in patients with coronary artery disease and heart failure, and a randomised study comparing optimum medical treatment to optimum medical treatment with complete revascularisation has just commenced in the United Kingdom (HEART UK) and will provide guidance regarding diagnosis and treatment of these patients. PMID- 12766497 TI - Coronary artery surgery for ischaemic heart failure: the surgeon's view. AB - Recent advances in medical therapy have improved outcomes for patients with severe heart failure. However, overall survival remains poor. Transplantation is an established therapy for these patients but is limited by the large mismatch between demand and donor organ availability. Recently it has been recognised that not all ventricular dysfunction secondary to coronary artery disease is irreversible. Revascularisation in certain patients would appear to improve ventricular function. These patients are said to demonstrate myocardial "hibernation". Revascularisation in these patients may provide a further treatment option in the treatment of heart failure. PMID- 12766498 TI - The role of echocardiography in the diagnosis and management of heart failure. AB - Heart failure has emerged as one of the most pressing health care issues in the United States. It is estimated that 4.8 million people have chronic heart failure, and approximately 400,000 new cases are diagnosed each year. Since the incidence of heart failure increases significantly with age, its prevalence is likely to increase as the population grows older. The American Heart Association and American College of Cardiology developed clinical practice heart failure guidelines to assist physicians in the diagnosis and management of patients with heart failure, in the hopes of reducing hospitalizations and mortality. These guidelines emphasize the importance of echocardiography in the management of acute and chronic heart failure. However, the guidelines do not elaborate on all the potential applications of echocardiography for this condition. This review was undertaken to examine in detail, the role of echocardiography in the initial management and long-term follow-up of patients with heart failure. PMID- 12766499 TI - Mechanisms of monomorphic ventricular tachycardia in coronary artery disease. PMID- 12766500 TI - Slow:fast and slow:slow AV nodal reentry in the rabbit resulting from longitudinal dissociation within the posterior AV nodal input. AB - OBJECTIVE: The anatomic and electrophysiologic bases for multiple forms of sustained AV nodal tachycardia were determined in the rabbit. METHODS: Intracellular microelectrode recordings were used to identify antegrade and retrograde conduction limbs of sustained tachycardias observed in 23 of 152 superfused rabbit AV junctions. RESULTS: Slow:slow tachycardias (196 +/- 12 msec cycle length) with nearly equal AH and HA intervals (99 +/- 12; 97 +/- 11 msec, respectively) and early atrial activation near the coronary sinus os were observed in 14 preparations and slow:fast tachycardias (189 +/- 11 msec cycle length) with an AH > HA interval (141 +/- 12; 48 +/- 10 msec, respectively) and early atrial activation along the anterior limbus of the fossa ovalis were observed in 11 preparations. Both tachycardias were associated with longitudinal dissociation and localized reentry within the triangle of Koch. Slow:fast and slow:slow tachycardias exhibited counterclockwise and clockwise reentry circuits, respectively. Both circuits were present in two preparations. Slow:fast AV nodal reentrant tachycardias could be reset with stimuli introduced near the coronary sinus os and the anterior AV nodal input. Slow:slow tachycardias could be reset only by stimuli introduced near the coronary sinus os. The fraction of the tachycardia cycle length contained within the compact AV node was greater for slow:fast (0.35 +/- 0.07) than slow:slow reentry (0.15 +/- 0.05, p = 0.026), suggesting a longer lower common pathway for slow:fast tachycardia. CONCLUSIONS: Longitudinal dissociation within the posterior AV nodal input incorporating the AV node can provide the reentrant substrate for two different clinical forms of sustained AV nodal tachycardias. PMID- 12766502 TI - Utility of patient-activated cardiac event recorders in the detection of cardiac arrhythmias. AB - BACKGROUND: Patient-activated event recorders are useful for the diagnosis of arrhythmia in patients with palpitation and presyncope. However, the utility of event recorders in patients suspected of arrhythmia but presenting with other symptoms is not clear. Furthermore, the factors influencing their utility have not been evaluated. METHODS: Event recorder reports of six hundred and sixty consecutive patients referred due to clinical suspicion of cardiac arrhythmia were reviewed. We divided symptoms into four groups: palpitation, presyncope, chest tightness, and dyspnea. We calculated the diagnostic yield according to patients' symptoms reported on the record, and analyzed the factors affecting the utility of event recorders. RESULTS: The overall diagnostic yield was 64%, and those of palpitation, presyncope, chest tightness, and dyspnea were 66%, 57%, 51% and 60%, respectively. The most common five findings in our patients were sinus rhythm (36%), sinus tachycardia (30%), atrial premature complex (14%), ventricular premature complex (12%), and atrial fibrillation/atrial flutter (7%). The recording duration (8 +/- 4 days) was short in the present study, but the overall diagnostic yield was similar to those of previous ones. Women had lower diagnostic yields than men, especially in atrial flutter-fibrillation. The diagnostic yield was not influenced by age, ordered doctors, or history of cardiovascular disease. CONCLUSIONS; Patient-activated event recorders provided a good diagnostic yield in patients with different presentations of cardiac arrhythmia, and women had lower diagnostic yield in atrial flutter-fibrillation. PMID- 12766501 TI - The results of pacing trials for the prevention and termination of atrial tachyarrhythmias: is there any evidence of therapeutic breakthrough? AB - Atrial fibrillation (AF) is now recognized as the most prevalent sustained cardiac arrhythmia and it is associated with considerable mortality and morbidity. The demand for effective therapeutic strategies for AF has always been high and is anticipated to further increase. Anticoagulation and pharmacologic antiarrhythmic therapy or radiofrequency catheter ablation remain the mainstay of treatment for AF. Among the wide range of nonpharmacologic options which are presently being investigated, only ablation in or around the pulmonary veins and the surgical maze procedure have been shown to accomplish the aim of the curative treatment of the arrhythmia. Preventative atrial pacing and antitachycardia pacing may offer an attractive alternative option for the management of AF by either eliminating the triggers and/or by modifying the substrate of the arrhythmia. The results of several recent trials have shown a significant increase in the time to first AF recurrence, a decrease in atrial conduction time, and a trend towards reduction in AF burden, suggesting that atrial pacing may prevent AF due to improved synchronization of atrial depolarization. The recognition of potential triggers of AF, such as atrial premature complexes, short-long sequence, and bradycardia, has encouraged the development of novel atrial pacing algorithms designed to prevent the initiation of the arrhythmia on an individual basis. Observations of AF often starting with regular atrial activity consistent with atrial tachycardia have supported the hypothesis that early antitachycardia pacing may prevent progression to AF. The concept of "hybrid therapy" based on the combination of several different therapeutic strategies suggests that antitachycardia pacing therapy, integrated with an atrial defibrillator and preventative atrial pacing modes, may act synergistically to prevent AF. Dual chamber cardioverter-defibrillators with capacity to prevent and interrupt AF may offer more comprehensive and successful treatment for patients with advanced heart disease, frequent paroxysms of AF, and the risk of proarrhythmia. PMID- 12766503 TI - Conduction velocity around the tricuspid valve annulus during type 1 atrial flutter: defining the location of areas of slow conduction by three-dimensional electroanatomical mapping. AB - BACKGROUND: Conduction velocity (CV) around the tricuspid valve annulus (TVA) during type 1 atrial flutter (AFL) has been shown to be slowest in the tricuspid valve-inferior vena cava (TV-IVC) isthmus, compared to the septal or free wall segments of the TVA. However, fiber orientation in the triangle-of-Koch suggests that the inferior septum and medial TV-IVC isthmus should be the most slowly conducting segments around the TVA. METHODS: To test this hypothesis we evaluated CV around the TVA during type 1 atrial flutter in 11 patients, using an electro anatomical mapping system (Carto). CV was first calculated in 4 segments around the TVA including the TV-IVC isthmus, lateral free wall, superior free wall and septum, and then calculated in 8 segments around the TVA including medial (MI) and lateral isthmus (LI), inferior (IL) and superior lateral (SL) free wall, lateral (LS) and medial superior (MS) free wall, and superior (SS) and inferior septum (IS). Statistical comparison of CV from these multiple segments was made by one-way analysis of variance. RESULTS: Measured in 4 segments around the TVA, mean CV (m/sec) in the TV-IVC isthmus (0.81 +/- 0.23) and the septum (0.93 +/- 0.18) was significantly slower than CV in the lateral free wall (1.16 +/- 0.23) and superior free wall (1.10 +/- 0.20), and CV in the TV-IVC isthmus was significantly slower than in the septum (p < 0.05). However, when analyzed in 8 segments, mean CV in the MI (0.56 +/- 0.16) and IS (0.59 +/- 0.24) was significantly (p < 0.05) slower than in all other segments including the LI (1.06 +/- 0.46), IL (1.17 +/- 0.40), SL (1.15 +/- 0.40), LS (1.04 +/- 0.25), MS (1.15 +/- 0.28), and SS (1.26 +/- 0.36) segments. CONCLUSIONS: Consistent with previous reports, CV around the TVA during type 1 AFL was slowest in the TV-IVC isthmus, compared to the septum, superior and lateral free wall regions. However, when the TVA was further subdivided into 8 segments, CV in the MI and IS segments was significantly slower than in all other segments around the TVA. These observations more precisely define the regions of slow conduction in human type 1 AFL, and are consistent with the known anisotropy and slow conduction in the Triangle of Koch. PMID- 12766504 TI - Directions of atrial excitation wavefront influenced antegrade conduction of manifest accessory atrioventricular pathway: a case report. AB - We report a case of left accessory atrioventricular pathway (AP) which antegrade conduction was manifest during pacing from the anterior site of the atrial insertion, but absent during pacing from the posterior site. Infusion of adenosine triphosphate during pacing from the posterior site induced a conduction block of atrioventricular node without affecting the persistent antegrade conduction block of the AP. These findings suggested that the different response of the AP according to the directional change of atrial conduction was not due to a "linking" phenomenon, but may relate to the impedance mismatch because of an oblique fashion of the pathway. PMID- 12766505 TI - Acute and chronic effects of cryoablation of the pulmonary veins in the dog as a potential treatment for focal atrial fibrillation. AB - BACKGROUND: Atrial fibrillation (AF) may be triggered by premature atrial depolarizations originating in the pulmonary veins (PV). Radiofrequency catheter ablation of PV foci may prevent recurrence of AF, but may cause PV stenosis. Therefore, a safer method for ablation of PV foci is needed. This study evaluated the acute and chronic effects of PV ablation using a cryocatheter ablation (CCA) system, which may be less likely to cause PV stenosis. METHODS: CCA was performed by freezing for 5 minutes or more in one or more PVs in 10 anesthetized dogs. Pacing threshold and vessel diameter were measured before and after PV cryoablation. All dogs were restudied at 4.0 +/- 1.64 months (range 2-7) in a manner identical to baseline. RESULTS: CCA was performed in 27 PVs (range 1 4/dog), with a mean freeze time of 8.62 +/- 5.42 minutes per vein (range 5.23 22.06). Mean temperature for all freezes was -65 +/- 5.3 degrees C. Mean PV diameter was 6.49 +/- 1.73 vs 6.24 +/- 1.83 mm (p = NS) and mean pacing threshold 1.32 +/- 0.75 vs 9.36 +/- 5.93 mA (p <.01), before vs. acutely after ablation. At followup, at the ablation sites PV diameter (7.02 +/- 1.88 mm) was unchanged from baseline, whereas pacing threshold remained elevated (2.54 +/- 1.44 mA, p <.05 vs baseline). There were no acute or long-term complications. CONCLUSIONS; (1) CCA of PVs produced a significant rise in acute and chronic pacing threshold indicating loss of atrial conductivity. (2) CCA of PVs did not cause PV stenosis or other complications. (3) The data suggest that CCA of PVs may be a safe and effective method for treating focal AF. PMID- 12766506 TI - Initial experience with a novel focused ultrasound ablation system for ring ablation outside the pulmonary vein. AB - INTRODUCTION: Atrial fibrillation has been shown to initiate from triggers within pulmonary veins. Several studies have documented that electrical isolation of those triggers can lead to maintenance of sinus rhythm. The complication of pulmonary vein stenosis has limited the utility of delivering ablation energy within the pulmonary vein. We utilize a focused ultrasound catheter ablation system for delivery of transmural ablation lines proximal to the pulmonary vein ostium. METHODS: Nine dogs (weight 30-39 kg) were anesthetized and ventilated. Through a transseptal approach, pulmonary veins were engaged with the focused balloon ultrasound catheter. Ultrasound power was delivered at 40 acoustic watts outside the pulmonary vein ostium, focused 2 mm off the balloon surface, with a depth of approximately 6 mm, for 30-120 seconds. Following ablation, lesions were histopathologically analyzed. RESULTS: Of nine animals studied, fourteen pulmonary veins were ablated. We found successful delivery of near circumferential and transmural ablation lines in 6/14 pulmonary veins. In each of the six circumferential ablations, successful alignment of the ultrasound transducer along the longitudinal axis of the parabolic balloon occurred. The final four ablations were conducted with an enhanced catheter design that assured axial alignment. Of these ablations, all four were circumferential. The remaining 8 pulmonary veins had incomplete delivery of lesions. In each of these veins the ultrasound transducer was misaligned with the balloon axis when therapy was delivered. CONCLUSION: Focused ultrasound ablation is a new means of performing pulmonary vein isolation. This method provides delivery of lesions outside the vein, limiting the risk of pulmonary vein stenosis for the treatment of atrial fibrillation. PMID- 12766508 TI - Ivan Mahaim (1897-1965). PMID- 12766507 TI - Large tip electrodes for successful elimination of atrial flutter resistant to conventional catheter ablation. AB - The most widely accepted criterion for successful radiofrequency catheter (RFC) ablation of typical atrial flutter is the development of bi-directional isthmus block. In a subset of patients, conventional RFC ablation fails to achieve this endpoint because deeper and wider lesions are required. We investigated the efficacy of a long 8-mm tip catheter in these cases. One hundred and seventy-four consecutive patients (137 male; 61 +/- 9 years) with recurrent typical atrial flutter underwent conventional RFC ablation first with a standard 4 mm tip catheter. In resistant cases (n = 52), ablation was continued using a large tip 8 mm catheter when the 4-mm tip catheter failed. Resistant atrial flutter was identified when 21 RFC pulses failed to reach the selected endpoint of bi directional isthmus block or in cases of transient bi- directional block (at least 3 episodes). In 122 of the 174 patients (70%) conventional atrial flutter ablation was successfully performed with 13 +/- 5 RFC applications. In the remaining 52 subjects (30%), the ablation procedure was completed using the large tip electrode catheter. In 30 of these 52 patients (58%), the catheter was changed because of persistent intra-atrial conduction after 21 RFC pulses and in 22 (42%) because of intermittent conduction block after 11 +/- 5 applications. Using the large tip electrode catheter, the selected endpoint was achieved in all patients of both groups with 3 +/- 2 RFC pulses (power output of 50-60 W, pulse duration of 60 sec). No post-procedure complications were observed. After 15 +/- 5 months of follow-up, 16 patients (9%) had recurrence of atrial flutter. Five of the patients had been in the resistant group. In patients with atrial flutter resistant to conventional ablation therapy, the long tip (8-mm) catheter appears to be a safe and effective alternative to use of the conventional 4-mm tip catheter. PMID- 12766509 TI - Worldwide utilization of implantable cardioverter/defibrillators now and in the future. AB - Despite the demonstrated efficacy of implantable cardioverter defibrillators (ICDs) in reducing sudden and total mortality in selected patients, their implantation rates vary greatly between countries. In the United States, implantation rate is 228 implants per million inhabitants compared to only 45 implants per million in Western Europe. The differences in the ICD use may be explained by following factors: manner in which sudden cardiac death is perceived by politicians and physicians (sudden cardiac death is perceived as a "nice way of dying"); differences in indications; physicians' information; prevalence of coronary artery disease; sudden cardiac death survival rates; perceived reliability of alternative treatment (namely antiarrhythmics including amiodarone); economic backgrounds; and health care politics. Furthermore, the cost of this treatment strategy must be considered. This issue has been raised because generalization of ICD use in patients matching clinical characteristics of patients enrolled in the primary prevention trials may represent a significant economic burden to be added to the already overloaded health care system. This low acceptance may not be entirely related to budget constraint but also to the perceived efficacy of ICDs by physicians and health authorities. PMID- 12766510 TI - Utilization of implantable defibrillators in Africa. AB - Sub-Saharan Africa is dominated by diseases of poverty. HIV/AIDS affects 28.5 out of a total of 600 million in the region. South Africa is the only country in sub Saharan Africa in which implantable cardiovertor defibrillators (ICDs) are implanted (0.8/million in 2001). Only 3 of the 35 new ICDs were implanted in state-funded public hospitals. The pacemaker implantation rate for South Africa was 41/million in 2001. Approximately 20% of the population consume 56% of the health care expenditure, mainly funded by Medical Insurance. A tax-funded state health care system serves the rest of the population, but is concentrated on improving sanitation and primary health care. Diversion of funds from academic tertiary hospitals has reduced specialised services, particularly cardiology and cardiac surgery, and has resulted in an exodus of skilled personnel to the private sector. In the rest of sub-Saharan Africa, tertiary health care is mainly privately funded. Cardiology and cardiac surgery is not widely available. Many countries are crippled by debt and chronic local conflicts. Only one state hospital (Groote Schuur, Cape Town) provides an electrophysiology (EP) service including catheter ablation and ICD implantation, and training in EP, by two electrophysiologists. EP services are available privately in 3 centres. No EP service exists in the rest of sub-Saharan Africa. PMID- 12766511 TI - The promise of resynchronization therapy. Who (and how many) will benefit? AB - It has been estimated that about 320,000 to 400,000 patients in the USA alone are possible candidates to cardiac resynchronization therapy according to the recently published AHA/ACC/NASPE guidelines for pacing and the results of the COMPANION trial. The selection of the most suitable candidate for CRT/CRTD is a crucial issue, but still a matter of debate. A large variety of clinical, invasive and non-invasive criteria have been proposed for appropriately selecting candidates for CRT. However, in all the studies the parameters have been retrospectively identified and none has reported their results in the form of a multivariate regression model. We have now well characterized the patients in sinus rhythm who most likely benefit from this non-pharmacological approach. The fact that the COMPANION trial was able to single out a specific subgroup of heart failure patients that can be treated better than what was very short time ago best medical therapy validates the large body of research that investigators worldwide have created about this therapy. Finally, the concept that any patients that require ventricular pacing, who have heart failure class II/III or IV may benefit from receiving biventricular rather than right ventricular pacing as much as the other patients with more classical indication for CRT is still open to discussion and needs to be tested in a randomized multicenter trial. PMID- 12766512 TI - Resynchronization therapy: implications for pacemaker implantation in Europe. AB - Cardiac resynchronization has proved a valuable adjunctive therapy in heart failure patients with ventricular conduction disturbances. It improves the patient's clinical status and quality of life, increases functional capacity and causes left ventricular reverse remodeling. The application of this new therapeutic method is expected to affect the pacemaker implantation rate, although the exact degree cannot yet be calculated. One of the main determining factors is the number of patients who are candidates for resynchronization therapy. Although this cannot be estimated precisely, it seems that it does not exceed 10-14% of heart failure patients. Additional factors that will influence the implantation rate, at least during the next few years, are the familiarization of cardiologists with the new implantation technique and follow up, the availability of cardiac laboratory time and the cost of this new therapy. Although the latter seems high, the financial burden will be minimal when it is taken into consideration that it will lead to a decrease in the number of hospital admissions and a significant reduction in the number of hospitalization days, the greatest determining factors in the current high management cost of heart failure patients. Further to the above, taking the differences between the implantation rates of conventional pacing systems into account in different European countries, the same could be expected of biventricular systems. Thus, although the incidence of heart failure is constantly increasing and conduction disturbances are frequent in such patients, we believe that the implantation rate of biventricular systems in Europe will not exceed an average of 10 per million population, fluctuating according to the country. In conclusion, resynchronization therapy has been established in special subsets of heart failure patients. Due to the epidemiological characteristics of heart failure, the implantation rate is expected to rise. Possible education and/or training issues that appear will be addressed respectively, enabling all suitable patients to enjoy the benefits this new therapeutic technique ensures. PMID- 12766513 TI - Impact of expanding indications on the safety of pacemakers and defibrillators. AB - The number of patients living with a pacemaker or implantable cardioverter defibrillator (ICD) increased markedly during the 1990's. Expanding device indications and increasing device sophistication has important implications for patient and device safety. While the FDA is responsible for assessing the safety and efficacy of medical devices, manufacturers, physicians, patients, medical professional societies, and legislators will all play a role in ensuring that medical device patients continue to receive safe, quality medical care. PMID- 12766514 TI - Legal implications of pacemaker and defibrillator guidelines. PMID- 12766515 TI - Malpractice liability implications of pacemaker and defibrillator guidelines in Canada. AB - Different guidelines for implantable pacemaker and cardioverter defibrillator therapy pose problems for legal liability in Canada. Medical practices in the United States provide measures for comparative assessment, but the Canadian health care system is operated and publicly funded on West European principles. Provincial governments accept the responsibility to provide their residents with reasonable access to medically necessary services, including devices, on conditions of partial reimbursement by federal government funds. However, some provinces may fund services for which they will not be reimbursed. Practitioners may practise according to governmental funding arrangements. The law recognizes that practitioners in a respected minority in their specialty can depart from generally approved professional guidelines, by retaining earlier practices that are not discredited, or by adopting newer, but non-mainstream practices. They can similarly follow professional guidelines different from those of their colleagues. Practitioners who depart from guidelines they profess to follow risk legal liability, but compliance does not invariably ensure judicial approval. Judges preserve the right to set standards of professional proficiency, since they are a matter of law, but usually endorse professional guidelines. Patients' choices must be adequately informed, according to well-established criteria of disclosure that must be applied to evolving professional views of best and acceptable practice, which are usually influenced by practice guidelines. Inadequate governmental provision of devices or services to ensure patients' care according to practice guidelines can be challenged in legal proceedings. Success may require governments to improve supplies, but judges may decline to give directions with serious budgetary implications. However, unsuccessful litigants against, for instance, governments or physicians, are liable to be required to pay the successful defendants' legal costs, and usually their own. PMID- 12766516 TI - New device indications: impact on the individual electrophysiology (EP) practitioner in a predominantly managed care environment. AB - The high penetration of managed care as a payment source in cardiology practice is a fact of life in many areas of the country. Understanding the system of reimbursement used by managed care is helpful in order to maximize physician payment for work done, to minimize practice overhead, and to reduce physician and managerial frustration. The key ingredient for success in any cardiology practice environment is high quality procedural outcomes and good communication with patients and referring physicians. This is true in both a managed care and fee for service environment. This article describes the relationship between an indicated electrophysiology procedure (e.g., an implantable cardioverter defibrillator in a low ejection fraction coronary patient) at one end and its reimbursement by a local managed care payer to an electrophysiologist at the other end. PMID- 12766517 TI - Expanding indications for defibrillators after myocardial infarction: risk stratification and cost effectiveness. AB - Practice guidelines have expanded to include a new Class IIa recommendation for implantable cardiac defibrillator (ICD) use in patients post-myocardial infarction (MI) with a left ventricular ejection fraction (LVEF) 0.12 seconds or LVEF 80% of all samples, whereas ethyl laurate and palmitate were detected in >50% of the samples. Ethyl myristate was the FAEE most commonly detected (>80%). All six meconium samples with confirmed maternal drinking histories tested positive for FAEE at significantly higher levels (mean, 11.08 nmol/g). The use of 2 nmol total FAEE/g meconium as the positive cutoff, when lauric and myristic acid ethyl esters were excluded, yielded the greatest sensitivity (100%) and specificity (98.4%). The authors conclude that certain FAEE are present at measurable levels in the meconium of neonates not exposed to maternal drinking, and correction is needed to allow high specificity. PMID- 12766553 TI - Population pharmacokinetics of carbamazepine in Chinese epilepsy patients. AB - AIM: To investigate the pharmacokinetic profile of carbamazepine (CBZ) in Chinese epilepsy patients. MATERIALS AND METHODS: Serum samples through concentrations at steady state (n = 687) were collected prospectively from 585 patients during routine clinical care. Data were analyzed by the non-linear mixed-effect modeling (NONMEM) technique with a one-compartment model of first-order absorption and elimination. RESULTS: The important determinants of clearance (CL) were total body weight (TBW); dose; patient age over 65 years (E); and comedication with phenytoin (PHT), phenobarbital (PB), or valproic acid (VPA) when VPA daily dose was greater than 18 mg/kg. The final pharmacokinetic model for relative CL and apparent distribution volume (V) were: Equation CONCLUSION: A population pharmacokinetic model was proposed to estimate the individual CL for Chinese patients receiving CBZ in terms of patient's dose, TBW, and comedications to establish a priori dosage regimens. PMID- 12766554 TI - Effects of various CYP2D6 genotypes on the steady-state plasma concentrations of risperidone and its active metabolite, 9-hydroxyrisperidone, in Japanese patients with schizophrenia. AB - The effects of various CYP2D6 genotypes on the steady-state plasma concentrations (Css) of risperidone and its active metabolite, 9-hydroxyrisperidone, were studied in 85 Japanese schizophrenic patients (27 men and 58 women) treated with 6 mg/d risperidone for at least 2 weeks. Plasma concentrations of risperidone and 9-hydroxyrisperidone were measured using liquid chromatography-tandem mass spectrometry. The patients had the following CYP2D6 genotypes: wild-type (wt)/wt (40 patients), CYP2D6*10 (*10)/wt ( 28), CYP2D6*5 (*5)/wt ( 8), *10/*10 ( 5), *5/*10 ( 3), and CYP2D6*4/CYP2D6*14 ( 1), respectively. The Css values of risperidone and 9-hydroxyrisperidone were corrected to the median body weight of 58 kg. The medians (ranges) of the Css of risperidone in the aforementioned genotype groups were 2.2 (0.37-35.7), 6.4 (2.1-26.5), 12.3 (4.7-39.5), 19.4 (13.4 26.4), 64.0 (41.6-68.8), and 91.8 nmol/L. Those values for risperidone-to-9 hydroxyrisperidone ratio were 0.03 (0.01-0.33), 0.06 (0.03-0.19), 0.14 (0.07 0.29), 0.28 (0.25-0.38), 0.48 (0.38-0.58), and 2.35, respectively. The Css of risperidone was significantly (P < 0.05 or P < 0.001) different among the four genotype groups (wt/wt, *10/wt, *5/wt, and *10/*10), except between the *5/wt and *10/*10 groups. Also, the risperidone-to-9-hydroxyrisperidone ratio significantly (P < 0.005 or P < 0.001) differed among these genotype groups. No significant differences were found in the Css of 9-hydroxyrisperidone and the active moiety (the Css of risperidone plus 9-hydroxyrisperidone) among these genotype groups. This study confirms previous findings that the CYP2D6 status affects the Css of risperidone via its strong regulation of 9-hydroxylation of risperidone. However, similar active moiety of risperidone among different genotype groups suggests that the determination of the CYP2D6 genotype has little importance for clinical situations. PMID- 12766556 TI - Adverse herbal interactions causing hypotension. AB - A 57-year-old man consulted an herbalist for epigastric discomfort. Four hours after he drank a decoction made from 14 herbs, he developed nausea, epigastric pain, and dizziness. He also had two loose bowel movements. On arrival at the hospital 4 hours later, his blood pressure was 77/46 mm Hg, and his pulse was 60 beats/min. He was given intravenous fluids. In the next 3 hours, his blood pressure gradually returned to his usual level of 100/65 mm Hg. His other gastrointestinal symptoms gradually subsided during the next 24 hours. His white cell count was 17.8 x 109/L but was normal on recheck. Complete cell counts, renal function and liver function tests, and electrocardiogram were otherwise normal. He was discharged home on day 2. Seven of the 14 herbs taken by this patient are known to have vasodilatory or blood pressure-lowering effects, and 3 of these herbs are used to manage hypertension. In traditional Chinese medicine, practitioners often use a combination of herbs in an attempt to improve the efficacy but reduce the adverse effects of treatment. The risk of adverse herbal interactions will also be higher. PMID- 12766555 TI - Hepatic venous stenosis-related reduction of cyclosporin clearance in a pediatric patient after liver transplantation. AB - A 20-month-old girl with Alagille syndrome developed stenosis of hepatic venous anastomosis after living related-donor liver transplantation. Cyclosporin (a microemulsion formulation) was given orally at a dose of 50 mg twice daily, and the trough blood concentration was relatively stable. Before balloon angioplasty of the hepatic vein, trough cyclosporin blood concentrations became elevated and consequently, the dosage was reduced to 15 mg twice daily. On the day of angioplasty, the calculated elimination rate constant of cyclosporin was 0.036 hr(-1), while its apparent basal value was 0.078 hr(-1). The cyclosporin trough concentration to dosage (C/D) ratio gradually increased reaching the maximum on the day after angioplasty. Thereafter, the C/D ratio promptly decreased. Thus, it is speculated that the increase in cyclosporine C/D ratio was mainly dependent on reduction of hepatic clearance of cyclosporin due to hepatic congestion caused by the stenosis of the hepatic venous anastomosis. PMID- 12766557 TI - Evaluation of tricyclic antidepressant false positivity in a pediatric case of cyproheptadine (periactin) overdose. AB - CASE REPORT: The authors report a case of pediatric cyproheptadine toxicity, initially misdiagnosed as tricyclic toxicity based on the results of a preliminary rapid toxicological serum screen. Although such cross-reactivity has been reported, the chemical basis of this observation has not yet been evaluated. By GC/MS methods and HPLC assays adapted for the detection of tricyclic compounds, the authors confirmed that cyproheptadine was indeed responsible for this patient's toxicity. In addition, the authors identified the presence of a cyproheptadine metabolite in the patient's serum. Further testing in an immunoassay-based toxicologic screen demonstrated some cross-reactivity exhibited by the patient's serum, but not the parent compound. These findings showed that the cross-reactivity correlated with the presence of the cyproheptadine metabolite, highlighting the value of confirmatory toxicologic testing of routine rapid toxicologic screens. PMID- 12766558 TI - Association of the CYP3A4*1B 5'-flanking region polymorphism with cyclosporine pharmacokinetics in healthy subjects. AB - To determine the relationship between CYP3A4*1B polymorphism and cyclosporine pharmacokinetic parameters among healthy volunteers, the oral cyclosporine pharmacokinetic study was performed in 14 healthy subjects. Blood cyclosporine concentrations were measured by a high performance liquid chromatography. Concentration-time data were analyzed by a non-compartmental method using WinNonLin, and the blood samples were genotyped for the CYP3A4*1B 5'-promotor region using the polymerase chain reaction and a restriction digest. Each cyclosporine pharmacokinetic parameter was compared using the one-way ANOVA test according to his or her CYP3A4*1B genotype. There were four (4) homozygous A/A (wild-type), four (4) homozygous G/G (variant) and six (6) heterozygous A/G genotypes for CYP3A4*1B in these 14 healthy volunteers. The mean AUC/D (ng.hr/mL/mg) of CsA were 21.5 +/- 6.0 (A/A), 11.7 +/- 3.2 (G/G) and 19.2 +/- 2.3 (A/G), P = 0.0103 and the mean CL/F (L/hr) were 49.4 +/- 13.9 (A/A), 83.5 +/- 16.0 (G/G), and 52.5 +/- 5.6 (A/G), P = 0.0024. All other parameters were not significantly different among the three genotypes. PMID- 12766559 TI - The effect of ethanol and pH on the adsorption of drugs from simulated gastric fluid onto activated charcoal. AB - The effect of ethanol and pH on the adsorption of acetaminophen (ACET), phenobarbital (PHB), phenytoin (PHY), salicylic acid (SA), and theophylline (THEO) from simulated gastric fluid onto activated charcoal was studied. For the ethanol study, each drug was prepared at a concentration of 10 g/L in ethanol; in hydrochloric acid (HCl), 1.2 mol/L; and in HCl, 1.2 mol/L, containing 75% ethanol, 50% ethanol, and 25% ethanol (v/v), respectively. For the pH study, each drug was prepared at a concentration of 10 g/L in HCl, 1.2 mol/L, pH 1.0, and in buffers of pH 1.7, 3.0, 4.0, 4.8, 5.8, 6.5, 7.4, and 9.4. After the addition of 1 g of activated charcoal to 10 mL of each solution, it was incubated for one hour at 37 degrees C. For comparison, in each experiment a blank consisting of the solution without charcoal was also incubated. With increasing concentrations of ethanol, there were substantial decreases in the adsorption of ACET, PHB, and PHY to charcoal. Ethanol-induced decreases in the adsorption of SA and THEO were less pronounced. Changes in pH did not affect the adsorption of ACET, PHB, PHY, or THEO. However, the adsorption of SA was decreased slightly at pH 1.0 and 3.0. PMID- 12766560 TI - Analysis of topiramate and its metabolites in plasma and urine of healthy subjects and patients with epilepsy by use of a novel liquid chromatography-mass spectrometry assay. AB - A novel liquid chromatography-mass spectrometry (LC-MS) method was developed and validated for quantification of topiramate (TPM) and its metabolites 10-hydroxy topiramate (10-OH-TPM), 9-hydroxy topiramate (9-OH-TPM), and 4,5-O desisopropylidene topiramate (4,5-diol-TPM) in plasma and urine. The method uses 0.5 mL of plasma or 1 mL of urine that is extracted with diethyl ether and analyzed by LC-MS. Positive ion mode detection enables tandem mass spectrometric (MS/MS) identification of the aforementioned four compounds. Calibration curves of TPM, 4,5-diol-TPM, 9-OH-TPM, and 10-OH-TPM in plasma and urine were prepared and validated over the concentration range of 0.625 to 40 microg/mL using TPM d(12) as an internal standard. Calibration curves were linear over this concentration range for TPM and its metabolites. Accuracy and precision ranged in urine from 83% to 114% and 4% to 13% (%CV), respectively, and in plasma from 82% to 108% and 6% to 13%, respectively. The applicability of the assay was evaluated by analyzing plasma samples from a healthy subject who received a single oral dose of TPM (200 mg) and urine samples from 11 patients with epilepsy treated with TPM (daily dose between 100 to 600 mg) alone or with other antiepileptic drugs. Only TPM was detected and quantified in the plasma samples, and its concentration ranged between 0.7 and 4.3 microg/mL. The concentrations of TPM and 10-OH TPM were quantifiable in all urine samples and ranged from 20 to 300 microg/mL for TPM and from 1 to 50 microg/mL for 10-OH-TPM. The metabolites 4,5 diol-TPM and 9-OH-TPM were also detected in all urine samples, but their concentrations were quantifiable only in 4 patients. An unidentified peak in the chromatograms obtained from patients' urine was attributed to 2,3-O desisopropylidene topiramate (2,3-diol-TPM). Due to a lack of reference material of 2,3-diol TPM and the similar MS/MS spectrum with 4,5-diol-TPM, the calibration curves of 4,5-diol-TPM were used for the quantification of its isomer 2,3-diol TPM. Based on these determinations, the apparent 2,3-diol-TPM-to-TPM concentration ratio in patients' urine ranged from 0.05 to 0.51 and the 10-OH-TPM to-TPM ratio ranged from 0.02 to 0.17. In conclusion, a novel LC-MS method for the assay of TPM and four of its metabolites in plasma and urine was developed. Its utilization for analysis of urine samples from patients with epilepsy showed that the method was suitable for analysis of TPM and its metabolites in clinical samples. Two quantitatively significant TPM metabolites (10-OH-TPM and 2,3-diol TPM) and two quantitatively minor metabolites (9-OH-TPM and 4,5-diol-TPM) were detected and quantified in urine samples from patients with epilepsy. PMID- 12766561 TI - Activated charcoal is effective but equilibrium dialysis is ineffective in removing oleander leaf extract and oleandrin from human serum: monitoring the effect by measuring apparent digoxin concentration. AB - Accidental poisoning from oleander leaf or oleander tea can be life threatening. The authors studied the effectiveness of activated charcoal and equilibrium dialysis in removing oleander leaf extract and commercially available oleandrin as well as oleandrigenin, the active components of oleander plant, from human serum. Oleander leaf extract was prepared in distilled water and drug-free serum was supplemented with the extract. Then serum was treated with activated charcoal at room temperature and an aliquot was removed at 0 minutes, 10 minutes, 20 minutes, and finally 30 minutes to study the presence of oleander extract by measuring the apparent digoxin concentration using the FPIA for digoxin. The authors observed effective removal of oleander extract by activated charcoal. When the authors supplemented other drug-free serum pools with pure oleandrin or oleandrigenin and then subsequently treated them with activated charcoal, the authors observed complete removal of digoxin-like immunoreactivity at the end of 30 minutes' treatment. When drug-free serum pool supplemented with either oleander leaf extract, oleandrin, or oleandrigenin was passed through a small column packed with activated charcoal, the authors observed almost no apparent digoxin concentration following the passage through the column indicating that activated charcoal is very effective in removing oleander from human serum in vitro. In contrast, when serum pools containing either oleander leaf extract or oleandrin were subjected to equilibrium dialysis against phosphate buffer at pH 7.4, the authors observed no significant reduction in apparent digoxin concentration even after 24 hours. The authors conclude that activated charcoal is effective but equilibrium dialysis is ineffective in removing oleander leaf extract from human serum. PMID- 12766562 TI - Doxorubicin-resistant, MRP1-expressing U-1285 cells are sensitive to idarubicin. AB - A doxorubicin-resistant subline (U-1285dox(900)) was derived from the human small cell lung carcinoma cell line U-1285. U-1285dox(900) was exposed to a wide range of anticancer agents to determine its resistance profile. In contrast to U-1285 cells, the resistant subline U-1285dox(900) expressed elevated MRP1 mRNA detected by reversed transcriptase-polymerase chain reaction (RT-PCR) and MRP1 protein analyzed with Western blot. Neither MDR1 mRNA nor P-glycoprotein could be detected in the parental cell line or resistant subline. U-1285dox(900) exhibited high resistance to doxorubicin, epirubicin, daunorubicin, and vincristine, an intermediate resistance to mitoxantrone, and a low resistance to etoposide. A collateral sensitivity to cytosine arabinoside, chlorodeoxyadenosine, and melphalan was observed. The resistance could be reversed by buthionine sulphoximine and verapamil for all tested drugs. Compared with daunorubicin, resistance to idarubicin was very low, 14-fold and 2.6-fold, respectively. This was associated with a higher accumulation due to a slower transport of idarubicin out of U-1285dox(900) cells. PMID- 12766563 TI - Reverse phase high-performance liquid chromatography method for the analysis of amprenavir, efavirenz, indinavir, lopinavir, nelfinavir and its active metabolite (M8), ritonavir, and saquinavir in heparinized human plasma. AB - The increasing interest in applying therapeutic drug monitoring (TDM) to antiretroviral therapy is related to the observed interindividual variation in antiretroviral pharmacokinetics that results in a wide range of drug exposure from fixed-dosing regimens and the rapid evolution in the availability of phenotypic assays that generate a target 50% inhibitory concentration (e.g., IC(50)) as a basis for adjusting individual antiretroviral dosages. To facilitate the application of TDM, a method for the simultaneous determination of eight species has been developed. This method is used to quantitate efavirenz and the following protease inhibitors: amprenavir, indinavir, lopinavir, nelfinavir and its active metabolite (M8), ritonavir, and saquinavir. The method using reversed phase high-performance liquid chromatography (RP-HPLC) was validated. Detection is effected using a photodiode-array detector (PDA) scanning at four different wavelengths. This method allows for detection of all analytes to a lower limit of quantitation of 0.1 to 0.2 microg/mL with an interday variation in CV ranging from 3.5% to 10.4%. The method is being applied to a TDM program that is currently being implemented in the authors' laboratory. PMID- 12766564 TI - Therapeutic drug monitoring of the newer antiepileptic drugs. AB - The aim of the present review is to discuss the potential value of therapeutic drug monitoring (TDM) of the newer antiepileptic drugs (AEDs) felbamate, gabapentin, lamotrigine, levetiracetam, oxcarbazepine, tiagabine, topiramate, vigabatrin, and zonisamide. Studies of the relationship between serum concentrations and clinical efficacy of these drugs are reviewed, and the potential value of TDM of the drugs is discussed based on their pharmacokinetic properties and mode of action. Analytical methods for the determination of the serum concentrations of these drugs are also briefly described. There are only some prospective data on the serum concentration-effect relationships, and few studies have been designed primarily to study these relationships. As TDM is not widely practiced for the newer AEDs, there are no generally accepted target ranges for any of these drugs, and for most a wide range in serum concentration is associated with clinical efficacy. Furthermore, a considerable overlap in drug concentrations related to toxicity and nonresponse is reported. Nevertheless, the current tentative target ranges for felbamate, gabapentin, lamotrigine, levetiracetam, oxcarbazepine (10-hydroxy-carbazepine metabolite), tiagabine, topiramate, vigabatrin, and zonisamide are 125 to 250 micromol/L, 70 to 120 micromol/L, 10 to 60 micromol/L, 35 to 120 micromol/L, 50 to 140 micomol/L, 50 to 250 nmol/L, 15 to 60 micromol/L, 6 to 278 micromol/L, and 45 to 180 micromol/L, respectively. Further systematic studies designed specifically to evaluate concentration-effect relationships of the new AEDs are urgently needed. Although routine monitoring in general cannot be recommended at present, measurements of some of the drugs is undoubtedly of help with individualization of treatment in selected cases in a particular clinical setting. PMID- 12766565 TI - Monitoring lithium in breast milk: an individualized approach for breast-feeding mothers. AB - Lithium is a drug of choice for the management of bipolar disorder, a disease frequently affecting women in their childbearing years. Unfortunately, this drug has typically been contraindicated in nursing women. Data in humans are limited with respect to the use of this drug in lactating women, and early reports suggest high excretion into milk. The purpose of this report was to verify the excretion of lithium into human milk and to assess infant safety after breast feeding. The authors found wide interpatient variability in lithium dose offered to the infant through breast milk (from 0% to 30% of maternal weight-adjusted dose), indicating that therapeutic drug monitoring of lithium in milk and/or in infant's blood, coupled with close monitoring of adverse effects, is a rational approach. Since therapeutic drug monitoring of lithium is routine, physicians caring for these women and infants should be encouraged to individualize their recommendations. PMID- 12766566 TI - Subtherapeutic antiretroviral plasma concentrations in routine clinical outpatient HIV care. AB - The objective of this study was to evaluate plasma concentrations of nonnucleoside reverse transcriptase inhibitors (NNRTIs) and protease inhibitors (PIs) within several dosing schemes in a cohort of HIV-infected patients in routine clinical practice and to find possible explanations for subtherapeutic plasma concentrations. Patients were included if a PI or NNRTI was part of their antiretroviral regimen, at least one plasma concentration was obtained, and a complete medication overview from community pharmacy records was available. The study period was from January 1998 to September 2001. Each plasma concentration was related to median plasma concentrations of a pharmacokinetic reference curve, yielding a concentration ratio (CR). A cutoff CR was defined for each antiretroviral drug per specific regimen, discriminating between >or=therapeutic and subtherapeutic concentrations. For the patients with subtherapeutic concentrations, it was sorted out whether drug interactions, adverse events and self-reported symptoms, or nonadherence could be the cause of the lower than expected plasma concentration. Ninety-seven HIV-infected patients fulfilled the criteria. During the defined period, 1145 plasma concentrations were available (median, 11; interquartile range, 8-14). Three hundred fourteen (27.4%) plasma concentrations were classified subtherapeutic. Drug interactions (2; 0.6%), adverse events and self-reported symptoms (67; 21.3%), and nonadherence (14; 4.5%) could only partly explain the subtherapeutic drug levels. Consequently, a large number of the subtherapeutic plasma concentrations (73.6%) remained inexplicable. A high number of subtherapeutic plasma concentrations were observed. No clear causes were found; thus, corrective measures will be difficult to employ. Therefore, therapeutic drug monitoring (TDM) must maintain its crucial place in routine clinical care to be able to identify patients who need extra attention so that therapeutic plasma concentrations are achieved. PMID- 12766567 TI - Simultaneous isocratic HPLC determination of vigabatrin and gabapentin in human plasma by dansyl derivatization. AB - A rapid and low-cost assay for simultaneous vigabatrin (VGA) and gabapentin (GBP) determination is described that can be performed with simple HPLC instrumentation. The method involves derivatization of the primary amine group of VGA and GBP with dansyl chloride followed by isocratic separation (column: microBondapak C-18, 10 microm, 300 x 3.9 mm; mobile phase: 50 mmol/L NaH(2)PO(4) in 40% acetonitrile) at 50 degrees C and fluorometric detection (excitation and emission wavelength: 318 and 510 nm, respectively) of the fluorescent product, which is stable for at least 7 days. Correlation coefficients of the calibration curves are >0.999 with a lower limit of detection of 0.3 microg/mL. Between- and within-run coefficients of variation are below 4.5%, and assay time is 15 minutes. This method may be used for therapeutic drug monitoring in the case of GBP and to control patient compliance in the case of VGA. PMID- 12766568 TI - Unbound valproate fraction in plasma and subcutaneous microdialysate in steady state and after a single dose in humans. AB - The aim of the current study was to characterize the observed discrepancy between unbound plasma valproate (VPA) in single dose and steady state in humans. Unbound and total plasma VPA and subcutaneous microdialysate VPA concentrations were estimated in single dose (6 subjects, n = 33) and steady state (11 subjects, n = 110). Trough plasma samples from 14 patients with total VPA concentrations of 300 micromol/L and 14 patients with VPA concentrations ranging from 600 to 700 micromol/L were analyzed for the unbound VPA fraction and compared with the unbound VPA fraction in spiked plasma samples from healthy subjects containing similar total VPA concentrations. The unbound plasma VPA fraction was significantly higher (P < 0.001) in the steady-state group compared with the single-dose group. The unbound VPA fraction was significantly higher in steady state compared with spiked plasma samples at high and low total VPA concentrations (P < 0.001). The difference between microdialysate and unbound plasma VPA concentrations was significant in the steady-state group (P < 0.001), while no difference was observed in the single-dose group. The mean (+/- SD) subcutaneous microdialysate-to-unbound plasma ratio in the single-dose and steady state groups was 1.08 (+/- 0.401) and 0.74 (+/- 0.123), respectively. The ratio difference between the groups was significant (P < 0.001). The results of the current study show that unbound plasma fractions of VPA are consistently higher in steady state compared with single dose. Together with the finding of higher unbound VPA fraction in steady state compared with spiked plasma samples, these results provide indirect evidence of displacement of VPA from plasma proteins by product(s) of VPA biotransformation. In addition, subcutaneous microdialysate VPA levels were consistently lower than unbound plasma levels in steady state but not after single dose. The mechanisms underlying this observation need to be studied further. PMID- 12766569 TI - How suitable are currently used carbamazepine immunoassays for quantifying carbamazepine-10,11-epoxide in serum samples? AB - The pharmacologic activity of the metabolite carbamazepine-10,11-epoxide (CBZ-E) is similar to that of carbamazepine (CBZ). As a result, determining the CBZ + CBZ E concentration would offer a better correlation with therapeutic or toxic effects than measurement of the parent drug alone. However, the most upto-date CBZ immunoassays are not able to quantitatively measure CBZ-E. Trough serum concentrations of CBZ were measured in 116 patients either in monotherapy (n = 66) or in polytherapy with other antiepileptic drugs (n = 50) using the Dade Dimension and Roche Cobas Integra immunoassays. The results were compared with those obtained for CBZ + CBZ-E by high-performance liquid chromatography (HPLC). The Dade Dimension immunoassay gave a concentration-dependent CBZ-E cross reactivity for the metabolite levels present in the samples studied (mean, 1.41 mg/L; range, 0.0-7.6 mg/L), whereas the Roche Cobas Integra gave a negligible cross-reactivity. The results provided by the Dade Dimension immunoassay only presented a clinically significant difference with regard to the CBZ + CBZ-E value in the group of patients comedicated with valproic acid. The results of this immunoassay revealed a high correlation and small standard error of the estimate for the total group of patients with the composite results of parent drug + metabolite obtained by HPLC (r = 0.993, ma68 = 0.22 mg/L), with a regression line CBZ + CBZ-E = 1.09 CBZ(Dade Dimension) + 0.07. By using the Dade Dimension immunoassay, it is possible to make a clinically valid estimate of the CBZ + CBZ-E concentrations in patients treated with CBZ in monotherapy and polytherapy. However, other immunoassays with a lower CBZ-E cross-reactivity are not suitable for this estimate to be made at increased metabolite levels. PMID- 12766570 TI - A simple method to calculate cyclosporine dosage to obtain a target C2 drug level. AB - C(2) Cyclosporine (CsA) level, as a surrogate of area under the time concentration curve (AUC) 0-4 hours, is a good predictor of drug absorption and clinical outcome after kidney transplantation. It has been difficult to define the optimal C(2) level in the individual case and given the broad range of C(2) due to interindividual absorption variability it has been troublesome to determine the drug dose needed to obtain an expected C(2)-CsA concentration. In this study data of 16 stable renal and renal/pancreas recipients treated with prednisone, azathioprine, and CsA (Neoral) managed by C(0) level was examined. CsA concentrations at time 0 (basal), 2, 6, and 12 hours post CsA (Neoral) intake were determined the day of the study. A significant linear regression level was established between C(2) (but not C(0), C(6) and C(12)) and the dosage expressed as mg/kg/d (P = 0.0113, correlation coefficient r = 0.573018). Subsequently, another 27 renal transplant recipients were studied retrospectively and divided into three groups according to posttransplant period: 1 to 6, 7 to 12, and beyond 12 months after transplant. Equations derived from the relationship between C(2) and dose (mg/kg/d) were similar between the three groups and when compared with the first study. A formula obtained from the 27 patients in the whole posttransplant period (mg/kg/d = C(2) x 0.0010208 + 1.86125) was applied to patients of the first study obtaining a regression coefficient between actual and calculated CsA dose of 0.6145 (P = 0.01). A more accurate equation (P = 0.0001, r = 0.5925) was obtained by analyzing 145 C(2) determinations covering a period from 1 month to 8 years following transplant which gave a linear regression line defined by the equation C(2) x 0.001473 + 1.6673. This equation would permit the calculation mg/kg/d of CsA (Neoral) dose to obtain an expected C(2) level. The derived equation shown in this paper has a predictive value of 50% to 60% only, but can help to find adequate dosage in the presence of an inappropriate C(2) level. PMID- 12766571 TI - Simultaneous determination of the HIV drugs indinavir, amprenavir, saquinavir, ritonavir, lopinavir, nelfinavir, the nelfinavir hydroxymetabolite M8, and nevirapine in human plasma by reversed-phase high-performance liquid chromatography. AB - A reversed-phase high-performance liquid chromatography method for the simultaneous quantitative determination of the currently available HIV protease inhibitors amprenavir, indinavir, lopinavir, nelfinavir, ritonavir, saquinavir, the active nelfinavir metabolite M8, and the nonnucleoside reverse transcriptase inhibitor nevirapine in human plasma is described. The method involved liquid liquid extraction from plasma, followed by high-performance liquid chromatography with an OmniSpher 5 C18 column and ultraviolet detection set at a wavelength of 215 nm for the protease inhibitors and 280 nm for nevirapine. The runtime was 25 minutes. The assay has been validated over the concentration range of 0.05 to 30 mg/L for indinavir, nelfinavir, ritonavir, and saquinavir, 0.07 to 30 mg/L for amprenavir and lopinavir, and 0.05 to 15 mg/L for M8 and nevirapine. This method proved to be simple, accurate, and precise and is useful for the therapeutic drug monitoring of protease inhibitors and the nonnucleoside reverse transcriptase inhibitor nevirapine on a routine basis. PMID- 12766572 TI - On-line derivatization utilizing solid-phase microextraction (SPME) for determination of busulphan in plasma using gas chromatography-mass spectrometry (GC-MS). AB - Busulphan (Bu) is an alkylating agent used in preparative regimen before stem cell transplantation (SCT). Bu has a narrow therapeutic window, and underdosing or overdosing may have a fatal outcome for the patient. Therapeutic drug monitoring (TDM) combined with dose adjustment is currently used to optimize and individualize therapy with Bu. However, this approach is limited to centers with laboratory facilities. An automated and easy method for measurement of Bu plasma concentrations may facilitate TDM for Bu and thus improve the clinical outcome. A solid-phase microextraction (SPME) on line with gas chromatography (GC) and mass spectrometric detection to quantify Bu in human plasma samples was developed using in-vial derivatization. Bu was mixed with reagent in a 2-mL vial and shaken for 15 minutes at 80 degrees C; subsequently, the SPME fiber was immersed into the vial for 15 minutes. The fiber was washed in water for 10 seconds before injection. Several parameters influencing the extraction and recovery were studied, such as absorption and desorption times, the effects of the temperature on the reaction, and the shaking time on the derivatization yield. Carbowax divinylbenzene, polyacrylate, and polydimethylsiloxane fibers were tested. The carbowax-divinylbenzene fiber resulted in the highest recovery in plasma samples. The validation of the method showed a high chromatographic selectivity and a good sensitivity (LOQ = 20 ng/mL). Coefficient of variation for SPME was less than 15%. The results showed good correlation between Bu concentrations and response within the range of 40 to 2500 ng/mL (R2 = 0.999). The accuracy ranged from 94% to 106%. This is well in line with the international criteria for validation. The present method was applied to patient plasma. The obtained results were comparable with the results obtained from GC with electron capture detection. The authors conclude that this method has shortened the analysis time considerably and is fully automated, which benefits TDM of Bu in SCT patients. PMID- 12766573 TI - Automated determination of free mycophenolic acid and its glucuronide in plasma from renal allograft recipients. AB - Mycophenolic acid, the active moiety of mycophenolate mofetil, inhibits the enzyme inosine monophosphate dehydrogenase. The main metabolite, mycophenolic acid glucuronide, has no immunosuppressive effect. Reported protein bindings are 97% for mycophenolic acid and 82% for mycophenolic acid glucuronide. Considerable intraindividual and interindividual variability in mycophenolic acid pharmacokinetics has been observed. Data on the variability of mycophenolic acid free fraction in plasma are sparse but may be relevant when discussing whether therapeutic drug monitoring of this drug is warranted. The authors describe a fully automated method for the determination of free concentrations by dialysis across a membrane followed by concentration of the dialysate on a trace enrichment column and liquid chromatography. Total concentrations are measured by protein precipitation and direct injection on the trace enrichment column. Plasma concentrations as low as 6 ng/mL free mycophenolic acid and 1 microg/mL free mycophenolic acid glucuronide can be measured with between-day coefficient of variation less than 15% and 6%, respectively. Stability testing confirmed that plasma samples could be stored for 14 days at 4 degrees C or -20 degrees C and at room temperature for approximately 12 hours without significant changes in free concentrations. Predose total and free concentrations of mycophenolic acid and mycophenolic acid glucuronide were determined in 27 samples from stable renal allograft recipients treated with mycophenolate mofetil, cyclosporin, and steroids. Total concentrations ranged from 0.57 to 16.2 microg/mL mycophenolic acid and 36 to 199 microg/mL mycophenolic acid glucuronide. Free concentrations ranged from 13 to 210 ng/mL mycophenolic acid and 8 to 58 microg/mL mycophenolic acid glucuronide. The method presented here has been successfully applied to measure free mycophenolic acid and free mycophenolic acid glucuronide in clinical samples. Further investigations may provide important data to support the identification of principles and target ranges for the monitoring of mycophenolic acid in the immunosuppressive therapy of organ transplant recipients. PMID- 12766574 TI - Epithelioid angiosarcoma of the bone: a series of 10 cases. AB - The clinical and pathologic features of 10 epithelioid angiosarcomas of bone were analyzed. There were eight males and two females who ranged in age from 26 to 83 years (mean 62 years). Four tumors were solitary and six were multifocal. In two consultation cases, the submitted diagnosis was metastatic carcinoma. Microscopically, the tumor cells were arranged in solid and infiltrative sheets, and in most cases vascular channels or cystically dilated spaces were present. The neoplastic cells had abundant eosinophilic cytoplasm and large nuclei with open chromatin and prominent eosinophilic nucleoli. Intratumoral hemorrhage, neutrophilic infiltrates, and intracytoplasmic lumina were frequently present. All 10 tumors stained positive for one or more endothelial markers, with CD31 being the most sensitive marker. Seven cases stained positive for cytokeratin. Ultrastructural examination in three tumors confirmed their endothelial differentiation. In the absence of obvious vascular differentiation, abundant intratumoral hemorrhage and intratumoral neutrophils are useful ancillary morphologic features that may suggest a vascular origin. Six patients are dead of disease, one is alive with metastasis, and two patients are currently disease free. Epithelioid angiosarcoma of bone should be included in the differential diagnosis of epithelioid neoplasms of bone, and endothelial markers should be a part of their immunohistochemical analysis to avoid the misdiagnosis of a metastatic carcinoma because of the significant differences in the treatment and clinical outcomes of these entities. PMID- 12766575 TI - Relevance of vertical growth pattern in thin level II cutaneous superficial spreading melanomas. AB - Thin (10% of tumor cells) in 55 of 72 cases. However, two distinct patterns were recognized among those positive: diffusely dense (>75%) and sporadic (<75%). The former was further characterized by a consistency of Bcl-6+ tumor cell density at any given area, resembling the staining pattern of the GC or follicular lymphoma (FL) (GC/FL pattern), whereas the latter was, besides less dense population, by variable density from area to area. The GC/FL pattern was observed in 36% and 38% of gastric and intestinal DLBCLs de novo, respectively, but in none of the gastric H-ML. CD10 was positive in 12 of 71 cases (17%), all coexpressing Bcl-6. CD10+ tumors were more frequent in the intestinal (33%) than in gastric DLBCLs ( approximately 15%). Significantly, CD10 expression was observed in three gastric H-MLs, including one that displayed a distinct lymphoepithelial lesion. IN CONCLUSION: 1). tumors showing a diffusely dense pattern of Bcl-6 expression should be distinguished from those showing a sporadic pattern; for the former most likely represents the tumor of GC B-cell derivation, and the latter non-GC, including MALT lymphoma; 2). tumor of GC B-cell origin thus defined accounted for about one third of gastric as well as intestinal DLBCLs de novo but none of the gastric H-ML; and 3). CD10 expression can be seen in MALT lymphomas and should not be used as the marker for GC B cells. PMID- 12766584 TI - Metastatic tumors of the vulva: a clinicopathologic study of 66 cases. AB - Metastatic tumors involving the vulva are rare, with only a few series and case reports published in the English literature to date. In this study, we present the clinicopathologic features of 66 cases of metastatic tumors of the vulva seen at the University of Texas M. D. Anderson Cancer Center from 1944 to 2001. The patients' age ranged from 18 to 84 years (mean 54.8 years). The most common presentations were vulvar nodules or a mass (39 cases), pain (7 cases), and ulceration (5 cases). In 46.9% of cases, the primary tumor was of gynecologic origin, whereas in 43.9% of cases the primary tumor was of nongynecologic origin. The remainder had unknown primaries. The site most frequently involved by metastasis was the labium majus (44 cases: 18 on the right, 13 on the left, 6 bilateral, and 7 unspecified side). Thirty percent of the patients received chemotherapy as treatment for the metastasis, 27% received radiotherapy, and the rest received some combination of chemotherapy, radiotherapy, and surgery. Of the 60 patients with available follow-up, 52 died of disease within 1-81 months (median 7.5 months) from diagnosis of the metastasis. Metastatic tumors of the vulva are rare; however, the diagnosis of these tumors is facilitated by the knowledge of a preexistent malignancy and the lack of a mucocutaneous intraepithelial lesion. PMID- 12766585 TI - International variation in histologic grading is large, and persistent feedback does not improve reproducibility. AB - Histologic grading systems are used to guide diagnosis, therapy, and audit on an international basis. The reproducibility of grading systems is usually tested within small groups of pathologists who have previously worked or trained together. This may underestimate the international variation of scoring systems. We therefore evaluated the reproducibility of an established system, the Banff classification of renal allograft pathology, throughout Europe. We also sought to improve reproducibility by providing individual feedback after each of 14 small groups of cases. Kappa values for all features studied were lower than any previously published, confirming that international variation is greater than interobserver variation as previously assessed. A prolonged attempt to improve reproducibility, using numeric or graphical feedback, failed to produce any detectable improvement. We then asked participants to grade selected photographs, to eliminate variation induced by pathologists viewing different areas of the slide. This produced improved kappa values only for some features. Improvement was influenced by the nature of the grade definitions. Definitions based on "area affected" by a process were not improved. The results indicate the danger of basing decisions on grading systems that may be applied very differently in different institutions. PMID- 12766586 TI - Solitary versus multiple carcinoid tumors of the ileum: a clinical and pathologic review of 68 cases. AB - It is well known that small intestinal carcinoid tumors may occur as solitary or multiple lesions. However, the biologic significance of multiple carcinoid tumors has not been clearly defined. The purpose of this study was to compare the clinical and pathologic features and prognosis of patients with solitary versus multiple carcinoid tumors of the ileum. Sixty-eight patients, including 50 with solitary and 18 with multiple carcinoid tumors, were included in the study. Hematoxylin and eosin-stained slides from routinely processed tumor resection specimens of the ileum were evaluated for a variety of histologic features such as tumor size, depth of invasion, tumor stage, and venous, perineural, and lymphovascular invasion. Follow-up and clinical data, such as patient age, gender, presenting complaints, presence of synchronous or metachronous malignancies, and presence of the carcinoid syndrome, were obtained and the results were compared between the two patient groups. Fifty patients with solitary carcinoid tumors (male/female ratio, 27:23) and 18 patients with multiple tumors (male/female ratio, 7:11) were identified. Patients with multiple carcinoid tumors were significantly younger than patients with solitary tumors at the time of diagnosis (55 years vs 63 years, p = 0.006). There was a high association between multiple carcinoid tumors and the carcinoid syndrome (4 of 18 vs 1 of 50, p = 0.004) as compared with patients with solitary carcinoid tumors. There was also an association between tumor multiplicity and venous invasion, but this relationship was not statistically significant (p = 0.07). The follow-up period was similar for both groups (mean 36 months, median 26 months, range 1-139 months). A significantly higher proportion of patients with multiple carcinoid tumors were either alive with disease or died of disease (56%) compared with those with solitary carcinoid tumors (18%, p = 0.002), and this relationship persisted in multivariate analysis (p = 0.02). Overall, no significant differences were observed between these two patient groups with respect to other clinicopathologic features such as tumor size, depth of invasion, presence of distant metastases, lymphatic or perineural invasion, or presence of an associated malignancy (p >0.05). In conclusion, we found that patients with multiple carcinoid tumors are younger, have a significantly greater risk of developing the carcinoid syndrome, and have a poorer prognosis than patients with solitary tumors. PMID- 12766587 TI - Burkitt lymphoma arising in organ transplant recipients: a clinicopathologic study of five cases. AB - We report five cases of Burkitt lymphoma arising in organ transplant recipients. There were four men and one woman with a mean age of 35 years. All were solid organ recipients with three renal, one liver, and one double lung transplantation. The time interval between organ transplantation and lymphoma averaged 4.5 years. Patients typically presented with high-stage disease with generalized lymphadenopathy and bone marrow involvement. Histology showed classic Burkitt lymphoma or atypical variant/Burkitt-like morphology. C-MYC rearrangement, including three cases with immunoglobulin heavy chain and two cases with lambda light chain, and Epstein-Barr virus were detected in all the cases. Additional chromosomal abnormalities were present in two of three cases and p53 mutation was found in one of three cases. Aberrant genotype and phenotype were frequently encountered, including minor monoclonal or oligoclonal T-cell populations and undetectable surface immunoglobulin light chain expression. Four patients received antilymphoma regimens, with combination chemotherapy (three patients) and/or Rituximab (three patients), in addition to reduction of immunosuppression. All four patients achieved complete remission. We conclude that posttransplant Burkitt lymphoma represents a characteristic clinicopathologic entity and occurs later after transplantation. Genotypic and phenotypic aberrations are often present. Rituximab may be an effective alternative to conventional combination chemotherapy in the treatment of a posttransplant Burkitt lymphoma. PMID- 12766588 TI - Ductal carcinoma in situ of the breast: impact of pathology on therapeutic decisions. AB - The therapy of ductal carcinoma in situ (DCIS) is controversial but is being increasingly decided by pathologic evidence. Studies of the natural history of DCIS demonstrate that DCIS is very heterogeneous in its clinical behavior. As detailed in several reviews, studies that followed patients after biopsy alone indicate a great difference between the small noncomedo examples of DCIS and the larger comedo DCIS lesions. The currently available evidence from cases that have been treated by planned surgical excision without radiation therapy would indicate that noncomedo examples of DCIS have a low incidence of recurrence and may be adequately treated by this technique. In contrast, comedo DCIS lesions have a high propensity for recurrence despite excision and radiotherapy. This presentation will review the histopathology of DCIS and highlight the idea that we are currently in a state of transition in our understanding of DCIS. Studies supporting the stratification of DCIS by histologic pattern plus cytology and size will be contrasted with the rapidly disappearing classic posture that all DCIS is biologically similar and treatment options need not be stratified by the different subtypes or varieties of DCIS. PMID- 12766589 TI - Special types of invasive breast cancer, with clinical implications. AB - Analysis of invasive breast cancers by histopathology has generally revolved around the analysis of individual characteristics such as nuclear pleomorphism and/or gland formation (grading) or categorization by specific histologic patterns (special type carcinomas). Recognition of special types of invasive breast carcinoma allows for the identification of women with an extremely good prognosis often approaching or equaling that of the general population of the same age (with unique problems for medullary). The distinguishing features of special types of breast cancer are present in complete or classic form in the pure examples of these cases, and the features recognizing these types may be present in lesser degrees in many carcinomas of no special type or not otherwise specified in which they often have important clinical correlates other than strong indicators of survival, such as recognizing the diffusely invasive feature of lobular and lobular-like cancers. Many other features have been noted as contenders for types of breast cancer (such as squamous, cystic, clear cell, histiocytoid, and lipid rich). These terms are not discussed here because they have no special clinical correlates relevant to diagnosis or prognosis. It should be reemphasized here that combined histologic grade (Nottingham system favored) with nuclear, glandular, and mitotic evaluation is expected by most authorities for clarification of diagnosis in all but rare, pure medullary cancers. (5, 13, 14, 27) The Nottingham grade 11 interacts with nodal status to produce a very strong prognostic index. The mitotic count may be the most important part of the grading system, certainly for prognosis within 5 years of diagnosis. (22, 26) PMID- 12766590 TI - Benign breast disease and breast cancer risk: morphology and beyond. AB - Evidence from clinical follow-up studies has indicated that there is a relationship between the presence of histologically proven benign breast disease and breast cancer risk and that the level of risk varies according to the histologic category of benign breast disease. In particular, proliferative lesions without atypia are associated with a 1.5- to 2-fold increase in risk, whereas atypical hyperplasias are associated with a fourfold to fivefold increase in breast cancer risk. A number of clinical factors appear to modify the risk associated with these lesions, including the time since biopsy, menopausal status, and family history of breast cancer. Recent studies have begun to evaluate the potential role of biologic, molecular, and genetic markers in assessing breast cancer risk in patients with benign breast disease. Additional information derived from clinicopathologic follow-up studies, epidemiologic studies, and molecular and genetic studies will provide new insights into benign breast disease and breast cancer risk. PMID- 12766591 TI - Sentinel lymph nodes and breast carcinoma: which micrometastases are clinically significant? AB - Sentinel lymph node biopsy is changing surgical management of breast cancer and pathologic evaluation of lymph nodes. Although it has long been known that lymph nodes contain occult metastases, pathologists have not generally pursued their identification. Compared with level I-II axillary dissection, the reduced number of sentinel lymph nodes has made additional evaluation more attractive; however, the consequences of increased detection of micrometastases has not been fully explored or appreciated. National data suggest that the composition of traditional TNM stage groupings is changing, with a recent increase in node positive, stage II breast cancer, most likely the result of increased pathologic scrutiny. Clinical management of this new group of stage II patients is complicated by the lack of a historic prognostic comparison group because many of these patients would have been classified as stage I, node-negative in the past. Early outcome data in sentinel lymph node biopsy suggest no adverse outcome for patients with metastases no larger than 2.0 mm, a finding aligned with the current definition of micrometastasis. When sentinel lymph nodes are sliced at 2.0-mm intervals and totally embedded, the probability of identifying all metastases >2.0 mm is high. Using reasonable sampling strategies, minute metastases have a nearly equal chance of being missed or detected. New staging guidelines have established a lower limit for micrometastases and defined metastases no larger than 0.2 mm as isolated tumor cells or tumor cell clusters; nodes with isolated tumor cells will be classified as node negative (pN0) for stage grouping. Rigorous strategies designed to reliably detect single cells or small cell clusters in sentinel nodes remain time-intensive and cost prohibitive. PMID- 12766592 TI - The Nobel Prize and pathology. PMID- 12766593 TI - Schwannoma with rhabdomyoblastic differentiation: a unique variant of malignant triton tumor. AB - A 54-year-old woman presented with intractable perianal, bilateral buttock, and radiating thigh/calf pain. An MRI scan showed an intradural, contrast-enhancing, ovoid mass in the cauda equina region at L1-L2. At laminectomy, the ovoid mass arose from a nerve root and, intact, was gross totally resected. Histologically, the dominant pattern was that of schwannoma. One year thereafter, the symptoms recurred. An MRI scan demonstrated an irregular, heterogeneously enhancing tumor recurrence. A repeat laminectomy disclosed a large fleshy tumor involving multiple nerve roots. The lesion was subtotally resected and showed pluridirectional differentiation toward embryonal rhabdomyosarcoma, primitive neuroectodermal tumor, and rare malignant epithelial cells. Review of the original tumor disclosed only foci of embryonal rhabdomyosarcoma and primitive neuroectodermal tumor. Based upon available data regarding divergent differentiation in peripheral nerve sheath tumors, this is a unique, previously undescribed tumor demonstrating rhabdomyosarcomatous, primitive neuroectodermal tumor and scant epithelial differentiation in a schwannoma. In essence, it is a variant of malignant Triton tumor because of its origin in a tumor consisting of well-differentiated Schwann cells. It supports the contention that the Schwann cell is the source of a variety of heterologous elements in nerve sheath tumors. PMID- 12766594 TI - A malignant ovarian tumor with osteoclast-like giant cells. AB - Neoplasms that are composed focally, predominantly, or exclusively of osteoclast like giant cells admixed with variably pleomorphic mononuclear cells have been described in a wide variety of organs. In this report, we describe the case of a 76-year-old woman with an 8-cm tumor that appeared to be localized to the ovary, that was composed predominantly of diffusely distributed, bland-appearing osteoclast-like giant cells admixed with pleomorphic mononuclear cells, and that was not associated with an ovarian cystic neoplasm. Hemorrhage, large zones of necrosis, and a high mitotic index were the other characteristics of the tumor. Immunohistochemically, the mononuclear cells were strongly positive for vimentin and proliferating cell nuclear antigen and were negative for keratin AE 1/3, CAM 5.2, cytokeratin 7, epithelial membrane antigen, beta-human chorionic gonadotropin, desmin, smooth muscle actin, p53, leukocyte common antigen, S-100, inhibin, alpha-1-antichymotrypsin, and CD68. The osteoclast-like giant cells displayed immunoreactivity for CD68, vimentin, alpha-1-antichymotrypsin, and leukocyte common antigen only. Ultrastructurally, rare intercellular junctions were present between mononuclear cells, suggestive of an epithelial histogenesis. Less than a dozen ovarian lesions with the "giant cell" designation have been described, and most of these cases are thought to be analogous to the "sarcoma like" nodules or other such lesions that have a well-known association with ovarian cystic neoplasms. Our case, in contrast, did not have an easily identifiable epithelial component and demonstrated both an infiltrative border and vascular invasion. This is, to the authors' knowledge, the first detailed clinicopathologic description of such a case as an ovarian lesion. PMID- 12766595 TI - 18F-FDG imaging of malignant pleural mesothelioma: scientiam impendere vero... PMID- 12766596 TI - Applications of fluorodeoxyglucose positron emission tomography in the diagnosis of infection. AB - The purpose of this study was to assess the accuracy of fluorodeoxyglucose positron emission tomography (FDG PET) in diagnosing infection in a large population of patients and in a variety of clinical circumstances where the performance of conventional imaging modalities has been questioned. We retrospectively analysed 167 FDG PET scans obtained to evaluate 175 anatomical sites for the presence of infection. The major indications for the scans were (1) complicated orthopaedic hardware (n=97), (2) chronic osteomyelitis (n=56), and (3) other (n=14: six fever of unknown origin, three vascular grafts, and five soft tissue). We assessed the overall diagnostic accuracy of FDG PET for each of these indications. In addition, we further analysed this modality's effectiveness by grouping the scans into specific clinical situations. A final diagnosis was made on the basis of surgical pathology and clinical follow-up for a minimum of 6 months. The overall accuracy of FDG PET in evaluating orthopaedic hardware was 96.2% for hip prosthesis, 81% for knee prosthesis, and 100% in 15 patients with other orthopaedic devices. Among the patients in our sample suspected of having chronic osteomyelitis, the accuracy was 91.2%. FDG PET was inaccurate in three cases of fever of unknown origin and accurate in all vascular graft and soft tissue infections. In 49 patients with a clinically apparent soft-tissue infection, FDG PET was able to detect or exclude underlying osteomyelitis with an accuracy of 92.3%. Among the 23 patients who had recent orthopaedic procedures, FDG PET imaging was accurate in 87% of cases. It is concluded that FDG PET is a highly effective imaging modality in the assessment of patients with suspected infection. PMID- 12766597 TI - Prediction of functional recovery after coronary bypass surgery using quantitative gated myocardial perfusion SPECT. AB - Previous studies have demonstrated that myocardial perfusion imaging using 99mTc tetrofosmin at rest allows viability assessment similar to that obtained with 201Tl imaging and 18F-fluorodeoxyglucose positron emission tomography (18F-FDG PET). The simultaneous assessment of perfusion and regional function is now available by quantitative gated myocardial perfusion single-photon emission computed tomography (SPECT). This study was designed to evaluate the utility of quantitative values of wall motion and wall thickening, calculated by quantitative gated myocardial perfusion SPECT, for the prediction of functional recovery after coronary bypass grafting (CABG). Fifty-six patients with coronary artery disease scheduled for CABG were included prospectively. All patients underwent 99mTc-tetrofosmin gated SPECT imaging at rest preoperatively and 3 months after CABG. The myocardium was divided into nine segments and the average quantitative values of regional perfusion (percentage uptake) (%), wall motion (mm) and wall thickening (%) were determined automatically using quantitative gated SPECT (QGS) software. The wall motion score was defined visually using a four-point scale (0, normal; 3, akinesis), and segments with severe asynergy (score of 2 or 3) with patent grafts were assessed. Of 77 segments with severe asynergy, 56 segments showed improved wall motion and 21 segments did not improve after CABG. The area under the receiver operating characteristic curve of wall thickening for the prediction of functional recovery was significantly higher (0.92) than that of the percentage uptake (0.77, P<0.017) or wall motion (0.60, P<0.0001). When each analysis used the optimal threshold, the wall thickening analysis (>or=10%) had a sensitivity of 95% and a specificity of 81%. These values tended to be higher than those of the percentage uptake (sensitivity, 86%; specificity, 67%). The wall motion analysis (>or=1.5 mm) had a significantly lower sensitivity of 75% and specificity of 43% than the wall thickening analysis (P=0.0038 and P=0.011, respectively). The results indicate that wall thickening, calculated by QGS software, may be more useful than regional perfusion or wall motion analysis for the prediction of functional recovery after CABG. The areas of asynergy with relatively preserved wall thickening may have the potential for improved function despite severely decreased perfusion. PMID- 12766598 TI - Evaluation of the response to therapy of head and neck squamous cell carcinoma by using 3-[123I]iodo-L-alpha-methyl tyrosine and single photon emission tomography. AB - Early detection of residual tumour tissue offers the possibility for rapid administration of adjuvant treatment. Single photon emission tomography (SPET) with 3-[123I]iodo-L-alpha-methyl tyrosine (IMT) offers the ability to detect recurrence. The aim of this study was to carry out a prospective evaluation of sequential IMT SPET before and after primary therapy and to determine the best timing for scanning in order to establish the response to treatment. Sixteen consecutive patients with histologically proven head and neck cancer (HNC), who underwent IMT SPET before therapy, within 1 week of therapy, and 1 and 3 months after completion of primary therapy were included. Images were classified, according to clinical evaluation, as indicating a high likelihood (HL), intermediate likelihood (IL) and low likelihood (LL) that residual tumoural tissue was present. The definitive clinicopathological diagnosis and follow-up was considered as the 'gold standard'. Based on the definitive clinicopathological outcome, 10 of 16 patients were diagnosed with evidence of local tumour and six without. Nine of 10 patients with evidence of local tumour presented with an HL IMT SPET image after 3 months, seven of whom were from within the first week. In this group, 1/10 patients was considered clinically HS the first week and eventually 4/10 patients became HL, of which there were three at 3 months. Of the six patients diagnosed without local evidence of tumour, with an average follow-up of 15 months, 6/6 were clinically LL in the first week. Three of six had a consistently LL IMT SPET from within the first week. The three other patients had an HL scan the first week, of which one became IL. It is concluded that IMT SPET assessed the response to primary therapy most accurately 3 months after completion of therapy. An IMT SPET image that indicates a high likelihood of residual tumoural tissue may allow earlier stratification of the patients for secondary treatment. If negative, an IMT SPET can exclude residual tumoural tissue from within the first week after completion of therapy. PMID- 12766600 TI - Lymphoscintigraphic details of sentinel lymph node detection in 82 patients with squamous cell carcinoma of the oral cavity and oropharynx. AB - Lymphoscintigraphy for sentinel node (SN) detection has been studied extensively in melanoma and breast cancer. In head and neck squamous cell carcinoma (HNSCC), however, experience in this field is relatively meagre. The purpose of this study was to document and evaluate lymphoscintigraphic findings in HNSCC patients. Eighty-two patients with clinical T1-T4 N0 SCC of the oral cavity or oropharynx received peritumoral injections of 25-75 MBq 99mTc-colloidal albumin (CA). Dynamic lymphoscintigraphy was performed in lateral projection for 20 min, followed by 2 min static imaging in anterior projection. In 26 patients, additional static images were obtained 2-6 h after injection of the tracer. In four of 82 patients, both early and late imaging revealed no tracer transport. In 78 of 82 patients, one (60), two (14) or three (4) SNs could be visualized, either by dynamic scintigraphy (73) or delayed static imaging (5). In 48 of 78 (62%) patients, the SN was visualized within the first minute of dynamic imaging. In particular, SNs of tumours of the mobile tongue were visualized within the first minute. No effect of T-stage or 99mTc-CA dose on the transport time of the tracer towards the SN was seen. The distribution of the SNs in the various levels of the neck relative to the primary tumour sites within the oral cavity was in concordance with the patterns of lymph node metastases reported traditionally for patients with SCC in the oral cavity. This study demonstrates the different variables affecting SN identification with lymphoscintigraphy using 99mTc-CA in HNSCC patients. PMID- 12766599 TI - Persistent vegetative state: evaluation of brain metabolism and brain perfusion with PET and SPECT. AB - Patients in persistent vegetative state (PVS) after severe head trauma were investigated with 99mTc-ECD SPECT and 18F-FDG PET to further characterize the degree of brain damage and to obtain insight into changes of brain perfusion and glucose metabolism. 18F-FDG PET and 99mTc-ECD SPECT were performed in 16 patients in PVS. Quantitative PET data were compared with that obtained from seven normal controls. After spatial normalization into Talairach space, global grey matter values and regional data using predefined ROI sets were derived. For comparison of PET and SPECT, regional data were normalized to their individual mean grey matter values. Patients in PVS showed significantly lower values of cerebral glucose metabolism than did the controls. The mean reduction of grey matter values in cortical and subcortical structures was 58%, except in the vermis cerebelli, where only a reduction of 16% was found compared to the controls. Comparing the glucose metabolism and perfusion within the patient group, the pattern of both modalities was similar in the neocortex and internal ganglia. In the cerebellar hemispheres a relatively higher perfusion than glucose metabolism was found. The overall reduction of 58% of glucose metabolism in grey matter structures is in accordance with other PET studies investigating PVS patients with different disease histories. The relative preserved activity of vermis cerebelli seems to be an uncommon finding not described by other authors up to now. PMID- 12766601 TI - The limitations of gallium scintigraphy in the differentiation between benign and malignant salivary gland mass lesions. AB - The aim of this study was to evaluate the ability of gallium scintigraphy to differentiate between benign and malignant salivary gland mass lesions and to identify what types of lesions surpass its diagnostic utility. By considering the uptake of 67Ga, 193 salivary gland masses were graded visually as negative, weakly positive, moderately positive or strongly positive in comparison to the uptake in the nasal cavity and the liver. The uptake was compared with histopathological findings. Among 39 malignant tumours, uptake was positive in 31 (79%) (strongly positive in 18, moderately positive in seven and weakly positive in six) and uptake was negative in eight (21%). Adenoid cystic carcinoma was the most common malignant tumour in our study (11/39), and uptake was negative in five (45%) of these tumours. Malignant tumours did not differ significantly in size despite differences in uptake. Among 154 benign lesions, uptake was negative in 101 (66%) and positive in 53 (34%) (strongly positive in 12, moderately positive in 19 and weakly positive in 22). Out of 88 pleomorphic adenomas, 41 (47%) showed positive uptake. Sensitivity, specificity and accuracy for gallium study were 80%, 66% and 68%, respectively, when the malignancy criterion was weakly positive uptake. Accuracy was greatest (83%) when the criterion was strongly positive uptake, but this criterion failed to detect more than a half of malignant tumours (46% sensitivity). In conclusion, gallium scintigraphy had limitations in differentiating between benign and malignant salivary gland mass lesions. Adenoid cystic carcinomas and pleomorphic adenomas were the principal sources of false negative and false positive results, respectively. PMID- 12766602 TI - Sentinel node detection using 99mTc-rhenium sulphide colloid in breast cancer patients: evaluation of 1 day and 2 day protocols, and a dose-finding study. AB - Sentinel node (SN) biopsy is a promising replacement for standard axillary lymph node dissection for the staging of early breast cancer, and various techniques have been studied to identify SNs with dye or radioactive colloid. This study assesses the effect of the dose of radioactivity and the time before biopsy in order to set standards for the use of 99mTc-rhenium sulphide for the detection of SNs in breast cancer patients. Sixty patients with stage T1-2 N0 M0 breast cancer underwent SN biopsy, which was immediately followed by standard axillary dissection to confirm the SN results. For SN biopsy, 99mTc-rhenium colloid was injected peritumorally. A 1 day (morning injection and afternoon surgery) or 2 day (day before afternoon injection and morning surgery) protocol was applied. A dose-finding study was performed simultaneously using 7.4-37 MBq for the 1 day protocol and 37-74 MBq for the 2 day protocol. A scintigram was taken at 2 h for the 1 day protocol and 16 h for the 2 day protocol. After the injection of blue dye, SN biopsy was performed with a gamma probe, followed by standard axillary node dissection. The radiation exposure received by the surgical team during the operation was monitored. Histopathological comparison between SNs and axillary nodes was performed. Patient characteristics that might affect the radiocolloid uptake by SNs were assessed. SNs were identified in all patients regardless of the dose or administration protocol used. Two patients showed false negative pathological SN results, and the negative predictive value was 96% and the positive predictive value was 100%. In addition, radiation exposure to the surgical team and the amount of radioactive surgical waste were low, especially at lower doses. Two groups of patient characteristics were related to SN uptake. One was the body mass index (BMI) and the other was the age or menopausal status. Patients with a larger BMI tended to take up a smaller amount of 99mTc colloid. Older or post-menopausal patients showed lower SN uptake. 99mTc-rhenium sulphide colloid is an efficient radiopharmaceutical for SN detection. Both 1 day and 2 day protocols have equally good efficacy, and the recommended dose is 7.4 MBq for the 1 day protocol and 37 MBq for the 2 day protocol. Patients with larger BMI and older or post-menopausal patients tend to take up less 99mTc colloid. PMID- 12766603 TI - Safe radiation exposure of medical personnel by using simple methods of radioprotection while administering 131I-lipiodol therapy for hepatocellular carcinoma. AB - The intra-arterial administration of 131I-lipiodol is a therapeutic approach increasingly used for the treatment of inoperable hepatocellular carcinomas. This technique has even become the reference treatment for hepatocellular carcinomas with portal thrombosis and is the only effective treatment to reduce the risk of recurrence among patients who could benefit from surgical operation. Currently, few data have been published concerning the levels of exposure for personnel carrying out this type of treatment. We undertook a dosimetric study targeted mainly on the exposure of the person performing the injection of 131I-lipiodol to show that this treatment can be carried out with an exposure at the extremities distinctly lower than the regulatory annual threshold by using simple means of radioprotection. The point of puncture was carried out at the level of left femoral artery, the preparation and injection of the therapeutic dose was carried out extemporaneously by the nuclear medicine specialist using a 10 ml syringe (for an injected volume of 4 ml) fitted with an adapted syringe protector. The injection was carried out as rapidly as possible under scopic control while avoiding reflux, with compression carried out by the radiologist. This study comprises 52 intra-arterial injections of 131I-lipiodol (2016+/-92 MBq). For the nuclear medicine specialists, 52 measurements were carried out at the level of the thorax and 41 on the fingers. For the radiologists, 22 measurements were carried out at the level of the thorax and six on their index fingers; nine measurements were carried out at the level of the thorax for the technologist and four at the level of the thorax for the stretcher bearer. For the nuclear medicine specialists, the average dose received at the level of the fingers varies between 140 and 443 microSv (according to the fingers) and the average dose at the thorax is 17 microSv. For the radiologists, the average dose received is 215 microSv at the level of the fingers and 15 microSv at the thorax. These results show that the administration of high therapeutic activities of 131I lipiodol can be carried out for the exposed personnel with a dose at the level of the fingers much lower than the European regulatory limit of 500 mSv. PMID- 12766604 TI - Evaluation of renal graft haemodynamia by 51Cr-EDTA and o-[131I]iodohippurate: its use in the early diagnosis of glomerular hyperfiltration. AB - Chronic rejection is the most important cause of renal graft dysfunction. Non immunological mechanisms have been suggested as a probable origin of chronic graft rejection, provoking a decrease in renal mass function, followed by glomerular hyperfiltration in the remnant nephrons, which could cause progressive glomerulosclerosis and functional loss. Early, or preclinical, identification of patients with glomerular hyperfiltration, defined as an increase in glomerular filtration fraction (GFF) and in glomerular capillary pressure (GCP), could prolong graft life. The objective of this study was to evaluate, non-invasively, stable renal graft haemodynamia and early glomerular hyperfiltration. We studied 116 renal transplant patients with stable renal function and five healthy living kidney donors with normal renal function. Glomerular filtration rate (GFR) and effective renal plasma flow (ERPF) were determined using 51Cr-EDTA and o [131I]iodohippurate, respectively. GFF was obtained from the relation between GFR and ERPF, and GCP from a mathematical model (Hall-Gomez' formula). A simultaneous analysis of renal function was performed. In transplant patients, the GFR and ERPF were significantly lower than in healthy, living, kidney donors (P<0.02). The same trend was observed for GCP (P<0.01), while GFF was not significantly different. Twelve patients (10.3%) had criteria of glomerular hyperfiltration. In patients without criteria of glomerular hyperfiltration, plasma level and clearance of creatinine were 128+/-33 micromol.l-1 and 56+/-15 ml.min-1, respectively; and in those patients with glomerular hyperfiltration criteria were 108+/-18 micromol.l-1 (P=NS) and 83+/-24 ml.min-1 (P=0.002) respectively. It is concluded that determinations of GFR, ERPF, GFF and GCP allow non-invasive evaluation of renal graft haemodynamia and can be useful in the early detection of glomerular hyperfiltration. PMID- 12766605 TI - Non-invasive adrenal imaging in primary aldosteronism. Sensitivity and positive predictive value of radiocholesterol scintigraphy, CT scan and MRI. AB - The aim of this study was to evaluate the sensitivity and positive predictive value (PPV) of dexamethasone-suppression norcholesterol scintigraphy (NCS), computed tomography (CT) scanning and magnetic resonance imaging (MRI) in patients with primary aldosteronism (PA) who had undergone unilateral adrenalectomy. A series of 49 patients with confirmed PA was reviewed. There were 18 (36.7%) men and 31 (63.3%) women, with a median age of 47 years (range, 23-66 years). NCS was performed in all patients, and 46 (93.9%) and 31 (63.2%) underwent CT scan and MRI, respectively. Final pathology showed an aldosterone producing adenoma in 45 (91.8%) patients, unilateral nodular cortical hyperplasia (NCH) in two (4.1%) and unilateral microscopic cortical hyperplasia (MCH) in two (4.1%). No aldosterone-producing carcinoma or bilateral adenomas were found. The greatest diameter of the removed adrenal tumour was in the range 8-40 mm (median, 14 mm). The PPV of adrenal imaging was 97.6% for NCS, 85.0% for CT scan (P=0.04) and 83.3% for MRI (P=0.03), and the sensitivity was 85.4%, 85.0% and 74.1%, respectively (P=NS). The age and the main biochemical parameters did not differ significantly (P=NS) between patients with true positive and false negative results of the imaging procedures. NCS accurately depicted all patients with NCH and MCH, whilst CT scan and MRI failed to diagnose such unilateral adrenal gland hyperfunction in two and three patients, respectively. The overall sensitivity of combined NCS and CT scan was 100%. In conclusion, in patients with PA, both NCS and CT scan are necessary to confirm the exclusive unilateral adrenal hyperfunction and, subsequently, to establish the appropriate treatment. PMID- 12766606 TI - 188Re-tin-colloid as a new therapeutic agent for rheumatoid arthritis. AB - Radiation synovectomy is a useful treatment modality in patients with refractory synovitis. We have developed a 188Re-tin-colloid as a new radiopharmaceutical agent and investigated its efficacy and safety in patients with rheumatoid arthritis. Radiation synovectomy was performed using 188Re-tin-colloid in 22 knees from 21 rheumatoid arthritis patients refractory to intra-articular corticosteroid injection. The efficacy and safety of administration of 370-1110 MBq of 188Re-tin-colloid were evaluated after 1, 3, 6, 9 and 12 months. Pain intensity on a visual analogue scale decreased significantly 12 months after therapy (mean+/-SD: 68.0+/-26.1 mm vs. 25.1+/-23.4 mm; P=0.0001 by the paired t test). Pain decreased in 19 cases (86.3%), joint tenderness improved in 14 cases (63.6%) and joint swelling was reduced in all cases (100%). 188Re-tin-colloid was safe. The residual activity of 188Re in the blood was 0.077%+/-0.25% of the injected dose. The radioactivity of 188Re in the urine was 0.14%+/-0.13% of the injected dose. Transient reactive synovitis was observed in 18 cases (81.8%). No clinical side-effects or abnormalities in leucocyte count, platelet count, liver function tests or urine analysis were observed in any patient. In conclusion, in this first study of radiation synovectomy using 188Re-tin-colloid for patients with rheumatoid arthritis, the treatment resulted in the improvement of arthritis and was well tolerated. PMID- 12766608 TI - Does diastolic function evaluated with radionuclide ventriculography predict mortality, hospitalization and the development of new onset heart failure? AB - Left ventricular systolic dysfunction (LVSD) in asymptomatic patients is associated with the development of heart failure (HF) and the degree of LVSD predicts prognosis. Whether left ventricular diastolic dysfunction (LVDD) predicts the development of HF or mortality is not known. Our objective was to investigate the predictive value of LVDD evaluated by radionuclide ventriculography (RN). All patients referred for RN during a 12 month period were included. Medical records were reviewed to determine characteristics of the patients at the time of RN and events occurring during a 5 year follow-up. Data from 195 patients were analysed. During the follow-up period 49 patients (25.1%) died, 41 (21.0%) were admitted to hospital and 25 (12.3%) developed HF. An ejection fraction (EF) <40% was associated with mortality (relative risk (RR), 2.04; P=0.001) and hospital admissions (RR, 1.33; P=0.002). Patients who developed subsequent HF had, on average, lower EF at baseline. In a multivariate analysis the lower the EF the more likely patients were to develop new onset HF (odds ratio, 0.92; 95% CI 0.88-0.97; P=0.003). LVDD evaluated with peak filling rate and time to peak filling rate was not associated with any of the outcomes. However, a higher proportion of patients with pre-existing HF at the time of the RN had abnormal LVDD than patients with no HF. LVDD evaluated by RN is associated with symptoms of HF at the time of assessment but is not a good predictor of mortality, hospitalization or new onset HF. EF remains a better predictor of outcomes. PMID- 12766607 TI - Investigations of 99mTc morpholino pretargeting in mice. AB - This laboratory is exploring the use of morpholinos (MORFs), synthetic DNA analogues, for nuclear medicine applications, including pretargeting. The anti CEA antibody MN14 was conjugated with an 18 mer MORF and with diethylenetriaminepentaacetic acid (DTPA) for 111In labelling. In a dual label pretargeting study, tumour-bearing nude mice received different doses of (MN14 DTPA-111In+MN14-MORF) followed, at various times after i.v. injection, by 0.15 microg complementary MORF (cMORF) radiolabelled with 99mTc via MAG(3). Animals were killed 3 h thereafter and tissues were counted for both radionuclides. The 99mTc-cMORF was also administered to tumour bearing mice that, 2 days previously, had received different doses of unlabelled MN14-MORF IgG or, as control, unlabelled Sandoglobulin IgG-MORF (Sandoz-MORF). Tumour uptake was higher at all time points for the labelled antibody itself versus labelled cMORF (8-10 vs 1.3 2.3%ID/g, respectively) in part due to the rapid clearance of cMORF through the kidneys. However, target to non-target ratios were superior for pretargeting at all time points and in all tissue except blood and kidneys. By pretargeting alone, these ratios were highest in all tissues for 15 microg compared to higher MN14-MORF dosages and in all cases were superior to that of the Sandoz-MORF control. The superior target to non-target ratios for pretargeting can be partially explained through calculations based on both radiolabels: after 24 h, only 0-6% of MORF on MN14 was bound by 99mTc-cMORF in liver and spleen suggesting that the antibody is sequestered in these organs and 'invisible' to labelled MORF. Fortunately, this was not the case in tumours in which 50-60% was bound. It is concluded that pretargeting using MORFs provided encouraging results in one mouse model/anti-tumour antibody system. The advantages of pretargeting in this model were evident in the superior target to non-target ratios obtained over conventional imaging. PMID- 12766609 TI - The interpretation of 131I scans in the evaluation of thyroid cancer, with an emphasis on false positive findings. AB - Radioiodine has aided the management of differentiated thyroid cancer for several decades. Most thyroid cancers retain the ability to trap iodine, and radionuclides of iodine can be used both diagnostically and therapeutically. The availability of sensitive diagnostic tests, coupled with the ability to deliver targeted therapy, gives physicians the ability to manage thyroid cancer better than with any other type of cancer. The correct interpretation of radioiodine scans is critical in the appropriate management of patients with thyroid cancer. False positive findings do occur. A radioiodine scan showing abnormal uptake outside the thyroid bed must be studied carefully and alternative reasons for the finding must be considered. The scan should be analysed systematically. Is there residual thyroid? If so, what is the 48 or 72 h neck uptake? Radioiodine uptake in the salivary glands, stomach, gastrointestinal and urinary tracts should be acknowledged as physiological. Diffuse uptake is seen in the liver in most patients with functioning thyroid at the time of their post-therapy scan. When there is uptake of the radioiodine outside these regions, contamination must be considered. A variety of cases illustrating true positive, true negative, and false positive findings is presented in this review, and the causes and consequences of misinterpretation of radioiodine scans are discussed. PMID- 12766610 TI - Molecular differentiation of high- and moderate-grade human prostate cancer by cDNA microarray analysis. AB - The prognosis of men with moderate-grade prostate cancer is uncertain. At present, there are few if any reliable molecular markers that can distinguish moderate-grade tumors from those that behave more aggressively. To better understand the molecular basis of human prostate cancer and potentially provide information toward more accurate prognosis, we measured and analyzed gene expression profiles of 13 high- and moderate-grade human prostate tumors using cDNA microarrays. The expression of 136 genes was observed to differ significantly (P < 0.001) between normal prostate and tumors using one-sample t testing and Wilcoxon ranking. Hierarchical clustering of genes demonstrated a relatively similar pattern of differential expression across the tumors. However, importantly, permutation t tests (two-tailed P < 0.001) revealed 21 genes whose expression profiles segregated moderate- and high-grade tumors from each other, which was significantly (P < 0.03) greater than what was expected by chance. These results were compared in silico with prostate cancer profiling efforts performed by other groups, including a meta-analysis of four data sets, which validated many of the dysregulated genes. We suggest that these data provide insight into the molecular nature of clinically aggressive prostate cancer. PMID- 12766611 TI - PTEN as a molecular marker to distinguish metastatic from primary synchronous endometrioid carcinomas of the ovary and uterus. AB - The distinction between two primary carcinomas on the one hand and a metastatic disease on the other hand in patients suffering from synchronous endometrioid carcinomas of the uterus and ovary is difficult. Exclusive histopathologic analysis appears to be insufficient and sometimes misleading. The tumor suppressor PTEN was found to be important in early neoplastic transformation in endometrioid carcinomas of the uterus. In this study, we screened synchronous endometrioid carcinomas of the uterus and ovary of 10 patients for loss of heterozygosity using seven different microsatellite markers at 10q23.3 and for mutations in the entire coding region of PTEN. Point mutations or microdeletions/insertions were found in six patients. Allelic loss at 10q23.3 was detected in eight patients. Based on conventional histology, a metastatic disease was diagnosed in seven patients and a concomitant uterine and ovarian carcinoma in three patients. After molecular analysis, the histopathologic diagnosis of three patients had to be revised. Histopathology represents the standard method to process tumor specimens from these patients. Nevertheless, mutation screen for PTEN and LOH analysis at 10q23.3 provide helpful genetic tools to establish a correct final diagnosis, which is important in view of prognosis and therapeutic implications. PMID- 12766612 TI - Laboratory strategies for efficient handling of paraffin-embedded tissues for molecular detection of clonality in non-hodgkin lymphomas. AB - We herein present a technical strategy to optimize DNA isolation from paraffin embedded tissue (PET). This includes the choice of adequate buffers for proteinase K digestion and multiplex PCR amplifications for assessing the appropriateness of DNA extracts for subsequent PCR assays for detecting clonality. We found that the association of proteinase K digestion in nonionic buffer and subsequent extract dilutions accounted for 79% of successful amplifications. A final efficiency of 88% was achieved by additional organic extractions and/or re-extractions. Comparisons were carried out with control DNA extracts from fresh samples to assess the efficiency of each clonality assay. Immunoglobulin CDRIII rearranged region amplification was more efficient for pregerminal center B-cell lymphomas in contrast to CDRII rearrangement detection, which was more effective for germinal and postgerminal lymphomas. T-cell clonality detection by TCRgamma PCR was less efficient in PET samples than in fresh tissues showing that DNA integrity is more critical for TCR than for IGH amplification. Two inconclusive cases without phenotypic markers and two other atypical lymphoproliferations masked by reactive T cells were diagnosed as plasmablastic lymphomas and as monoclonal B-proliferations, respectively, due to IGH rearrangements. PMID- 12766613 TI - Gastric cancer cell detection in peritoneal washing: cytology versus RT-PCR for CEA transcripts. AB - This study investigates the sensitivity and specificity of cytology, qualitative, and real-time RT-PCR methods in free cancer cell detection of peritoneal washing from gastric cancer patients. Peritoneal washings were collected from 65 gastric cancer patients for routine cytology and total RNA extraction for qualitative and real-time RT-PCR for CEA. The sensitivity and false-positive rate was 51.1%, 0% for cytology, 48.9% and 5% for qualitative RT-PCR for CEA, and 42.5% and 5% for real-time RT-PCR for CEA. The qualitative and real time RT-PCR results show high concordance rate (89.7%). The highest sensitivity was obtained by the combination of cytology with qualitative RT-PCR for CEA (70.2%). RT-PCR results were positive in 63.6% of cytologic "atypia" cases. Combination of cytology and either of the RT-PCR methods resulted in significantly higher sensitivity than any one of the three methods alone (P < 0.05). There was no definite advantage of the real-time RT-PCR over the conventional RT-PCR. PMID- 12766614 TI - Microsatellite instability and p53 expression in gallbladder carcinomas. AB - We studied the MSI (microsatellite instability) status and p53 expression in a series of 71 gallbladder cancers (GCs) of different histologic type. All neoplasms were examined combining a microsatellite analysis at mononucleotide locus BAT-26 and an immunohistochemical study for hMSH2, hMLH1, and p53 proteins and markers of gastric and intestinal differentiation. All the 71 GCs were MSS (microsatellite stable). The p53 protein was found in 100% of undifferentiated GCs, 67% of conventional gallbladder adenocarcinomas, 50% of mucinous adenocarcinomas, and 20% GCs with squamous differentiation. All 71 MSS tumors showed presence of immunohistochemical expression of both hMLH1 and hMSH2 gene products. We concluded that microsatellite instability does not play a role in the developing of GC while p53 seems to be the most important alteration found in a large proportion of these cancers, with the only exception of mucinous and squamous gallbladder carcinomas. PMID- 12766615 TI - Rapid diagnosis of smallpox infection and differentiation from its mimics. AB - The potential for a bioterrorism-induced smallpox outbreak has been much discussed of late. The literature of the late 1960s stressed that the distinction between smallpox and the other viral-induced vesicle-forming diseases, namely varicella zoster and disseminated herpes simplex, was difficult to make. Given that the cutaneous manifestations of smallpox would be among the initial symptoms, we reviewed 2 cases of smallpox diagnosed in South America in the 1970s in conjunction with 9 cases of multiple skin vesicles diagnosed as either disseminated herpes simplex or varicella-zoster. These were examined by routine hematoxylin and eosin stain (H&E) as well as by in situ hybridization. A blind review of the cases demonstrated that each showed striking intraepithelial vesicles containing multinucleated squamous cells exhibiting a ground glass appearance of the nuclear chromatin. Thus, as expected, routine H&E examination could not differentiate the 2 smallpox cases from the other 9 samples. In situ hybridization easily distinguished the 2 cases of smallpox from the other 9 samples, 5 of which contained varicella-zoster (two had been misdiagnosed as herpes) and the other 4 were disseminated herpes simplex. The in situ test, readily accomplished in any histology-based molecular laboratory in 4 hours, allows for the rapid and specific identification of smallpox infection and, importantly, its distinction from its mimics. Formalin fixation, which is optimal for in situ hybridization, guarantees the inactivation of the smallpox virus. PMID- 12766616 TI - Laser capture microdissection and real-time PCR for analysis of glomerular endothelin-1 gene expression in mesangiolysis of rat anti-Thy 1.1 and murine Habu Snake Venom glomerulonephritis. AB - Molecular analysis of pathologic changes in glomeruli requires methods allowing rapid and exact detection of alterations in gene expression. Here, we analyzed endothelin-1 (ET-1) mRNA expression in mesangiolytic glomeruli during the course of a rat and murine model of mesangioproliferative glomerulonephritis (GN). A novel method combining laser capture microdissection (LCM), which permits the precise removal of selected mesangiolytic glomeruli, with a highly sensitive real time RT-PCR technique was used. Anti-Thy 1.1. GN was introduced in male Sprague Dawley rats (1.0 mg/kg body weight of OX-7 IV) and Habu Snake Venom GN was introduced in C57BL6 mice (habu snake venom toxin 6 mg/kg body weight IV). The degree of mesangiolysis during both GNs was analyzed using a semiquantitative scoring system. Mesangiolytic glomeruli were microdissected at different days of the diseases (day 2, 6, and 12 in anti-Thy 1.1 GN and days 1, 3, 7, and 14 in Habu Snake Venom GN) and from normal control animals. After RNA extraction and cDNA synthesis, ET-1 gene expression was measured by real-time RT-PCR. In parallel, in anti-Thy 1.1. GN ET-1 mRNA expression was analyzed using semiquantitative nonradioactive in situ hybridization; ET-1 protein expression was investigated by immunohistochemistry. Mesangiolysis peaked at day 6 in anti Thy1.1 GN and at day 1 in Habu Snake Venom GN. Mesangiolytic glomeruli were easily microdissected on cryostat sections in both models; quantification of mRNA with RT-PCR was reliable and reproducible. Glomerular ET-1 mRNA expression increased during the course of anti-Thy 1.1 GN and Habu Snake Venom GN peaked when mesangiolysis was most pronounced. This was seen by RT-PCR after glomerular LCM and by in situ hybridization; in parallel, glomerular ET-1 protein expression was increased. Combination of LCM and RT-PCR is a reliable method for quantification of localized gene expression in isolated renal structures. The above data argue for an important role of ET-1 in pathogenesis and/or repair of mesangiolysis in experimental mesangioproliferative GN. PMID- 12766617 TI - A polymorphism in the alpha4 nicotinic receptor gene (Chrna4) modulates enhancement of nicotinic receptor function by ethanol. AB - BACKGROUND: Several studies indicate that ethanol enhances the activity of alpha4beta2 nicotinic acetylcholine receptors (nAChR). Our laboratory has identified a polymorphism in the alpha4 gene that results in the substitution of an alanine (A) for threonine (T) at amino acid position 529 in the second intracellular loop of the alpha4 protein. Mouse strains expressing the A variant have, in general, greater nAChR-mediated 86Rb+ efflux in response to nicotine than strains with the T variant. However, the possibility of the polymorphism modulating the effects of ethanol on the 86Rb+ efflux response has not been investigated. METHODS: We have used the 86Rb+ efflux method to study the acute effects of ethanol on the function of the alpha4beta2 nAChR in the thalamus in six different mouse strains. Experiments were also performed on tissue samples taken from F2 intercross animals. The F2 animals were derived from A/J mice crossed with a substrain of C57BL/6J mice that carried a null mutation for the gene encoding the beta2 nAChR subunit. RESULTS: In strains carrying the A polymorphism (A/J, AKR/J, C3H/Ibg), coapplication of ethanol (10-100 mM) with nicotine (0.03-300 microM) increased maximal ion flux when compared with nicotine alone with no effect on agonist potency. In contrast, ethanol had little effect on the nicotine concentration-response curve in tissue prepared from strains carrying the T polymorphism (Balb/Ibg, C57BL/6J, C58/J). Experiments with the F2 hybrids demonstrated that one copy of the A polymorphism was sufficient to produce a significant enhancement of nAChR function by ethanol (50 mM) in animals that were also beta2 +/+. Ethanol had no effect on nicotine concentration response curves in T/T beta2 +/+ animals. CONCLUSIONS: The results suggest that the A/T polymorphism influences the initial sensitivity of the alpha4beta2 nAChR to ethanol. PMID- 12766618 TI - Ethanol potentiation of glycine receptors expressed in Xenopus oocytes antagonized by increased atmospheric pressure. AB - BACKGROUND: Behavioral and biochemical studies indicate that exposure to 12 times normal atmospheric pressure (12 ATA) of helium-oxygen gas (heliox) is a direct, selective ethanol antagonist. The current study begins to test the hypothesis that ethanol acts by a common mechanism on ligand-gated ion channels by expanding previous hyperbaric investigations on gamma-aminobutyric acid type A (GABA(A)) receptors (GABA(A)Rs) at the biochemical level to alpha(1)glycine (GlyRs) expressed in Xenopus oocytes. METHODS: Oocytes expressing wild-type alpha(1) homomeric GlyRs were voltage-clamped (-70 mV) and tested in the presence of glycine (EC(2)) +/- ethanol (50-200 mM) under 1 ATA control and 3 to 12 ATA heliox conditions. Glycine concentration response curves, strychnine/glycine interactions, and zinc (Zn2+) modulation of GlyR function was also tested. RESULTS: Pressure reversibly antagonized the action of ethanol. The degree of antagonism increased as pressure increased. Pressure did not significantly alter the effects of glycine, strychnine, or Zn2+, indicating that ethanol antagonism by pressure cannot be attributed to alterations by pressure of normal GlyR function. The antagonism did not reflect tolerance to ethanol, receptor desensitization, or receptor rundown. CONCLUSION: This is the first use of hyperbarics to investigate the mechanism of action of ethanol in recombinant receptors. The findings indicate that pressure directly and selectively antagonizes ethanol potentiation of alpha(1)GlyR function in a reversible and concentration- and pressure-dependent manner. The sensitivity of ethanol potentiation of GlyR function to pressure antagonism indicates that ethanol acts by a common, pressure-antagonism-sensitive mechanism in GlyRs and GABA(A)Rs. The findings also support the hypothesis that ethanol potentiation of GlyR function plays a role in mediating the sedative-hypnotic effects of ethanol. PMID- 12766619 TI - Genetic analyses of ethanol-induced hyperglycemia. AB - BACKGROUND: Genetic sensitivity to ethanol-induced hyperglycemia was hypothesized to be related to sensitivity to ethanol-induced hypothermia and conditioned taste aversion. These hypotheses were explored by characterizing blood glucose changes after ethanol exposure in BXD recombinant inbred mice. METHODS: Adult male and female BXD recombinant inbred mice were acutely exposed to 4 g/kg of ethanol or saline with the order of exposure counterbalanced, and separated by a 1-week interval. Tail blood samples and rectal temperatures were determined immediately before ethanol/saline exposure and 2 hr after exposure. RESULTS: Substantial strain differences in ethanol-induced hyperglycemia and hypothermia were noted. In addition, sex also determined sensitivity to ethanol-induced hyperglycemia and interacted with strain. Correlational analyses using strain means indicated that ethanol-induced hyperglycemia was genetically independent from ethanol-induced hypothermia or conditioned taste aversion. Quantitative trait locus (QTL) analyses indicated provisional QTL for ethanol-induced hyperglycemia on chromosomes 1, 3, 4, 6, 7, 9, 12, 14, and 18, which, in part, were sex specific. CONCLUSIONS: These findings indicate important sex differences in the glycemic response to ethanol. In addition, multiple genes likely control this response, independent from genes that are important for the thermic or aversive effects of ethanol. PMID- 12766620 TI - EEG low-voltage alpha and alpha power in African American young adults: relation to family history of alcoholism. AB - BACKGROUND: Several studies support an association between having a low-voltage EEG and alcohol dependence; however, it is not clear whether this measure represents a risk marker or is a result of years of heavy drinking. The present study's aims were to investigate the prevalence of low-voltage alpha EEG variants in African American young adults who have not yet developed alcohol dependence and to test for associations between low-voltage alpha (LVA) EEG, alpha power, and a family history of alcohol dependence. METHODS: Clinical ratings and spectral characteristics of the EEG, collected using bipolar recordings, were investigated in 81 African American young adult men and women (18-25 years old) who had no personal history of alcohol dependence. Information on psychiatric diagnoses, personality features, personal drinking and drug use history, and family history (FH) of alcoholism was obtained. RESULTS: Thirty-two percent (n = 26) of the participants had an LVA EEG variant, and an additional 22% (n = 18) had borderline LVA. The presence of an LVA variant was not associated with drinking status, a family history of alcoholism, or a personal history of anxiety disorders but was associated with significantly higher extroversion scores. Participants who had an FH of alcoholism had significantly higher spectral power in the slow alpha frequencies (7.5-9 Hz). FH was not associated with any significant differences in any other EEG frequency band. CONCLUSIONS: These findings suggest that considerable ethic variation may exist in the prevalence of LVA EEG variants. In addition, like findings in other populations of non-African descent, having an FH of alcohol dependence may be associated with significantly higher voltage in the alpha frequency ranges. PMID- 12766621 TI - Alcohol affects executive cognitive functioning differentially on the ascending versus descending limb of the blood alcohol concentration curve. AB - BACKGROUND: Executive cognitive functioning (ECF), a construct that includes cognitive abilities such as planning, abstract reasoning, and the capacity to govern self-directed behavior, has been recently researched as an antecedent to many forms of psychopathology and has been implicated in alcohol-related aggression. This study was designed to examine whether differential ECF impairments can be noted on the ascending versus the descending limbs of the blood alcohol concentration curve. METHODS: Forty-one male university students participated in this study. Twenty-one subjects were given 1.32 ml of 95% alcohol per kilogram of body weight, mixed with orange juice, and the remaining 20 were given a placebo. Participants were randomly assigned to either an ascending or descending blood alcohol group and were tested on six tests of ECF on their assigned limb. Subjective mood data were also collected. RESULTS: Intoxicated participants on both limbs demonstrated ECF impairment; the descending-limb group showed greater impairment than their ascending-limb counterparts. Intoxicated subjects were significantly more anxious at baseline than placebo subjects. The introduction of this covariate nullified any significant differences in subjective mood found on either limb of the blood alcohol concentration curve, but ECF impairments remained robust. CONCLUSIONS: Our results support the conclusion that alcohol negatively affects cognitive performance and has a differential effect on the descending versus the ascending limb of the blood alcohol concentration curve. The latter finding may have important ramifications relating to the detrimental consequences of alcohol intoxication. PMID- 12766622 TI - Mecamylamine attenuates the subjective stimulant-like effects of alcohol in social drinkers. AB - BACKGROUND: Recent studies have implicated central nicotinic cholinergic receptor systems in the reinforcing properties of alcohol. In laboratory animals, mecamylamine, a central nicotinic receptor antagonist, reduces the consumption of and preference for alcohol. This study investigated the effect of mecamylamine on the subjective responses to alcohol in humans. It was hypothesized that mecamylamine (7.5 and 15 mg) would attenuate the stimulant-like subjective effects of alcohol (0.8 g/kg) and decrease the self-reported desire to consume additional alcohol beverages. METHODS: Fourteen male and 13 female nonsmokers participated in 6 laboratory sessions. During each session, subjects received, in randomized order under double-blinded conditions, a capsule containing mecamylamine (7.5 or 15 mg) or placebo followed by a beverage containing alcohol (0.8 g/kg) or placebo. Physiologic and subjective-effect measures were taken at 30-min intervals for 2 hr after beverage consumption. RESULTS: Mecamylamine attenuated the stimulant and euphoric effects of alcohol and reduced the self reported desire to consume additional alcohol beverages. This effect was most pronounced in men, even though women exhibited greater physiologic reactions to mecamylamine. CONCLUSIONS: These findings suggest that nicotinic cholinergic receptors are involved in mediating some of the stimulant-like effects of alcohol. PMID- 12766623 TI - Neuropeptide Y reduces oral ethanol intake in alcohol-preferring (P) rats following a period of imposed ethanol abstinence. AB - BACKGROUND: Intracerebroventricular infusion of NPY has been shown to reduce ethanol intake in alcohol-preferring (P) rats in a limited access procedure. The purpose of the present investigation was to extend this finding to a two-bottle free-choice continuous access procedure in groups of rats that either did or did not undergo a period of imposed ethanol abstinence and ethanol reinstatement. METHODS: In experiment 1, female P rats were given 6 weeks of continuous access to ethanol (8% w/v) and water. Ethanol was removed for a period of 2 weeks during which the rats were surgically implanted with a cannula into the lateral ventricle. Following the ethanol abstinence period and immediately before ethanol reinstatement, rats received a single infusion of either artificial cerebrospinal fluid or NPY (10 microg). Ethanol and water intake was measured at both 4 hr and 24 hr after infusion, and 24-hr intake measures were taken daily for 13 postinfusion days. Experiment 2 was run in parallel with experiment 1, with the exception that rats did not undergo a period of imposed ethanol abstinence. Also, food intake was measured 4 and 24 hr after infusion. RESULTS: Following 2 weeks of imposed ethanol abstinence (experiment 1), NPY suppressed ethanol intake through postinfusion day 2. After uninterrupted continuous access to ethanol (experiment 2), NPY suppressed ethanol intake to a lesser extent and this effect lasted only 24 hr. NPY increased food intake at the 4-hr but not the 24-hr measure. CONCLUSIONS: Previous findings that central administration of NPY suppresses ethanol intake in P rats are extended by this study to a continuous access procedure, and the effect is amplified following a period of imposed ethanol abstinence. This effect of NPY compares favorably to results obtained with other treatments tested in similar animal models and provides support for a role of NPY in an allostasis model of addiction. PMID- 12766624 TI - Subjective intoxication in response to alcohol challenge: heritability and covariation with personality, breath alcohol level, and drinking history. AB - BACKGROUND: Numerous studies have identified differences in subjective response to alcohol in subjects differentiated by family history of alcoholism. Results suggest that genetic influences on individual variation in subjective response to alcohol may be a mechanism for genetic effects on alcohol problems. However, direct evidence for genetic effects on subjective response to alcohol is very limited. METHODS: In a sample of 99 adult twin pairs, we studied genetic influences on subjective intoxication after alcohol challenge. The twins ingested a standard dose of ethanol (0.70 g/kg for men/0.65 g/kg for women), and two measures of subjective response to alcohol were assessed. RESULTS: Genetic effects on variation in subjective intoxication reported 1 hr after drinking were significant and substantial: heritability was 0.60 for a 22-item scale and 0.48 for a brief 2-item measure. Self-report measures of neuroticism, psychasthenia, hostility, and family problems shared significant genetic covariation with subjective intoxication. Achieved breath alcohol level, rate of change in breath alcohol on the descending limb, and individual drinking history all shared familial variation with subjective intoxication. No significant genetic effects for subjective intoxication were found 2 hr after drinking, but familial influences remained present, and many of the same personality, drinking history, and breath alcohol variables were predictive of intoxication. CONCLUSIONS: Subjective response to alcohol is heritable, and genetic effects on subjective intoxication are partly shared with genetic effects on personality. PMID- 12766625 TI - Alcohol choice and amphetamine effects in light and moderate drinkers. AB - BACKGROUND: The results of previously published reports suggest that light and moderate drinkers respond differently to the effects of commonly abused sedatives (e.g., diazepam or ethanol). The purpose of this experiment was to determine whether light and moderate drinkers respond differentially to the effects of ethanol and d-amphetamine. METHODS: In the first phase of this experiment, volunteers (eight light drinkers and eight moderate drinkers) randomly sampled 0.5 g/kg of ethanol and placebo across two separate sessions. In the second phase, volunteers completed three sessions in which they chose either ethanol or placebo. In the third phase, volunteers received 0, 5, 10, and 15 mg of d amphetamine. Each dose was tested twice. After drug administration in each phase, volunteers completed a battery of subject-rated, performance, and physiologic measures periodically for 5 hr. RESULTS: Ethanol produced prototypical subject rated effects (e.g., increased ratings on the Alcohol Sensation Scale), but it was chosen over placebo infrequently. Light and moderate drinkers did not differ in terms of the self-reported or reinforcing effects of ethanol. d-Amphetamine produced prototypical subject-rated stimulant-like effects (e.g., dose-dependent increases in ratings of High and Rush). Moderate drinkers reported significantly greater drug effects than light drinkers. Responses to ethanol reliably predicted subsequent responses to d-amphetamine on several measures. CONCLUSIONS: The results of this experiment suggest that even moderate ethanol use may increase an individual's vulnerability to abuse drugs. Future studies should determine whether light and moderate drinkers respond differentially to other commonly abused drugs (e.g., opioids) and whether behavioral responses to ethanol also predict responses to these compounds. PMID- 12766626 TI - Serotonin transporter gene variation is associated with alcohol sensitivity in rhesus macaques exposed to early-life stress. AB - BACKGROUND: Decreased sensitivity to alcohol has been demonstrated to be a predictor of alcoholism in humans, and variation in the gene-linked polymorphic region of the serotonin transporter (5-HTTLPR) is associated with the response to the motor-impairing effects of alcohol. In a nonhuman primate model of excessive alcohol intake, we have shown that decreased serotonin turnover is associated with both lower initial sensitivity to alcohol and higher prospective alcohol consumption using rhesus macaques. In addition, we have demonstrated that macaques separated from their mothers and reared in peer-only groups are more likely to consume alcohol as adults. METHOD: To examine the relationship between serotonin transporter genotype, early rearing experience, and initial sensitivity to alcohol, peer- and mother-reared, adolescent, alcohol-naive rhesus macaques (n = 123) were rated for intoxication after intravenous administration of ethanol (2.2 g/kg and 2.0 g/kg for males and females, respectively) during two testing periods. Serotonin transporter (rh5-HTTLPR) genotype was determined using polymerase chain reaction followed by gel electrophoresis, and data were analyzed using ANOVA and the Mann-Whitney U test. RESULTS: Our analyses demonstrate an effect of serotonin transporter gene variation on ethanol sensitivity, such that animals homozygous for the l allele exhibited decreased sensitivity to the ataxic and sedating effects of alcohol. This effect remained after correction for blood ethanol concentrations and birth cohort. When animals were segregated according to rearing condition, serotonin transporter gene variation predicted intoxication scores among peer-reared animals. CONCLUSIONS: As in some human reports, this study demonstrates a diminution in the response to alcohol in animals homozygous for the l rh5-HTTLPR allele. The phenotypic expression of this genotype in l/s animals, however, is environmentally dependent. PMID- 12766627 TI - A 5-year prospective evaluation of DSM-IV alcohol dependence with and without a physiological component. AB - BACKGROUND: The DSM-III-R removed tolerance and withdrawal as required elements for a diagnosis of alcohol dependence. Although this practice was continued in DSM-IV, the more recent manual asked clinicians to note whether physiological aspects of withdrawal (tolerance and withdrawal) had ever been experienced. Few studies have determined the prognostic meaning of a history of a physiological component to DSM-IV alcohol dependence. METHODS: Face-to-face structured interviews were used to evaluate the course of alcohol, drug, and psychiatric problems during the subsequent 5 years for 1094 alcohol-dependent men and women. These subjects had been classified into subgroups at the time of initial interview regarding evidence of tolerance or withdrawal, and all evaluations were based on DSM-IV criteria. At baseline, the application of DSM-IV diagnostic guidelines resulted in 649 (59.3%) individuals having a history of an alcohol withdrawal syndrome, with or without tolerance (group 1); 391 (35.7%) with histories of tolerance but not withdrawal (group 2); and 54 (4.9%) with no lifetime histories of tolerance or withdrawal (group 3). RESULTS: During the 5 year follow-up, both the broad (group 1 plus 2 versus group 3) and narrow (group 1 versus group 2 plus group 3) definitions of physiological dependence were associated with more alcohol and drug problems. However, for most items, this differential primarily reflected differences between groups 1 and 3, with a less impressive effect by group 2. Although no group differences were noted for the rate of independent major depressive episodes, substance-induced depressions did differentiate among groups, a finding also most closely related to the distinction between groups 1 and 3. CONCLUSIONS: These data support the prognostic importance of noting the presence of a physiological component in alcohol dependence and indicate the potential relevance of limiting the definition of a physiological component to withdrawal. PMID- 12766629 TI - Alcohol availability as a predictor of youth drinking and driving: a hierarchical analysis of survey and archival data. AB - BACKGROUND: Much attention has recently been directed toward developing preventive interventions to reduce drinking and driving through efforts to limit the numbers and locations of alcohol outlets at the community level. Although evaluations of these efforts have suggested linkages between alcohol outlets and problem outcomes, they have not addressed the linkage between outlets and drinking and driving among youth. The analysis reported here investigates the relationship between alcohol outlet densities and underage drinking and driving as self-reported on two telephone surveys conducted in California. METHODS: These analyses were based on data obtained from two telephone surveys conducted by the Prevention Research Center and archival data collected by the California Department of Alcoholic Beverage Control and the US Census Bureau. The sample for the first survey consisted of 15- to 20-year-old adolescents and young adults contacted by telephone, using a random digit dialing of exchanges in the greater San Francisco Bay Area. A second set of survey data was similarly collected by a random sample of households throughout California, and the Bay Area subset was also used for this analysis. RESULTS: At the individual level, older respondents were more likely to report drinking and driving and riding with drinking drivers, whereas females and Asians were less so. At the aggregate or city-level, alcohol outlet density, as measured by the number of on- and off-premises establishments licensed to sell alcohol, was associated with both drinking and driving and riding with drinking drivers. These effects were moderated by a number of individual level effects, with younger respondents and females more likely to be affected by outlet densities. CONCLUSIONS: The findings here provide support for the implementation of policies targeting alcohol outlet density reductions. Areas with large numbers of such outlets provide ample opportunities to youth for alcohol purchases. PMID- 12766628 TI - The reliability and validity of the self-reported drinking measures in the Army's Health Risk Appraisal survey. AB - BACKGROUND: The reliability and validity of self-reported drinking behaviors from the Army Health Risk Appraisal (HRA) survey are unknown. METHODS: We compared demographics and health experiences of those who completed the HRA with those who did not (1991-1998). We also evaluated the reliability and validity of eight HRA alcohol-related items, including the CAGE, weekly drinking quantity, and drinking and driving measures. We used Cohen's kappa and Pearson's r to assess reliability and convergent validity. To assess criterion (predictive) validity, we used proportional hazards and logistical regression models predicting alcohol-related hospitalizations and alcohol-related separations from the Army, respectively. RESULTS: A total of 404,966 soldiers completed an HRA. No particular demographic group seems to be over- or underrepresented. Although few respondents skipped alcohol items, those who did tended to be older and of minority race. The alcohol items demonstrate a reasonable degree of reliability, with Cronbach's alpha = 0.69 and test-retest reliability associations in the 0.75-0.80 range for most items over 2- to 30-day interims between surveys. The alcohol measures showed good criterion-related validity: those consuming more than 21 drinks per week were at 6 times the risk for subsequent alcohol-related hospitalization versus those who abstained from drinking (hazard ratio, 6.36; 95% confidence interval=5.79, 6.99). Those who said their friends worried about their drinking were almost 5 times more likely to be discharged due to alcoholism (risk ratio, 4.9; 95% confidence interval=4.00, 6.04) and 6 times more likely to experience an alcohol-related hospitalization (hazard ratio, 6.24; 95% confidence interval=5.74, 6.77). CONCLUSIONS: The Army's HRA alcohol items seem to elicit reliable and valid responses. Because HRAs contain identifiers, alcohol use can be linked with subsequent health and occupational outcomes, making the HRA a useful epidemiological research tool. Associations between perceived peer opinions of drinking and subsequent problems deserve further exploration. PMID- 12766630 TI - Age-dependent changes in electroencephalographic responses to alcohol consumption in subjects with aldehyde dehydrogenase-2 genetic variations. AB - BACKGROUND: We have recently reported that alcohol consumption resulted in a significant increase in alpha power of the EEGs in aldehyde dehydrogenase-2 (ALDH2)-normal (NN) subjects but not in ALDH2-deficient heterozygote (ND) subjects. The purpose of the present study was to investigate interactive effects of individual factors such as age and ALDH2 genotype on alcohol-induced EEG changes. METHODS: We examined EEG power spectral changes induced by 0.4 ml/kg of alcohol ingestion in 53 NN and 21 ND subjects of two different age groups: younger and older groups. Blood ethanol and acetaldehyde levels were also determined in 17 NN and 13 ND subjects in separate studies. RESULTS: Alcohol consumption markedly increased EEG power in the NN subjects of the older group, especially in theta and slow alpha power, whereas only slight increases were noted in fast alpha and beta power in the NN subjects of the younger group. However, no such differences between the two age groups were observed in the ND subjects. It should be noted that there were no differences in blood ethanol and acetaldehyde level at 30 min after alcohol ingestion between the different age groups in both genotypes. However, there was a significant increase in frequency of alcohol intake in the older group of both genotype groups. The multiple regression analysis indicated that both alcohol use habits and genotype, as well as aging, significantly modulated EEG changes after alcohol ingestion. CONCLUSIONS: The results suggest that both ALDH2 genotype and age as well as alcohol use habits modify alcohol sensitivity in the central nervous system, resulting in greater increases in EEG energy in response to alcohol intake in the older group of the NN subjects. PMID- 12766631 TI - Persistent alterations of vasopressin and N-terminal proatrial natriuretic peptide plasma levels in long-term abstinent alcoholics. AB - BACKGROUND: During alcohol withdrawal and early abstinence, severe alterations of electrolyte and water homeostasis and their regulating hormones are well recognized. Almost nothing is known about regeneration of these functions with long-term abstinence. This cohort study was designed to monitor determinants of electrolyte and water balance over 280 days of abstinence in alcohol-dependent men compared with healthy controls. METHODS: Vasopressin (AVP), N-terminal proatrial natriuretic peptide, aldosterone, angiotensin II, and electrolytes, together with major parameters of kidney and liver function, were monitored in 35 male alcoholics aged 44 +/- 8 years. Of these, 21 could be followed up to 280 days of strictly controlled abstinence due to their participation in the Outpatient Long-Term Intensive Therapy for Alcoholics. The control group comprised 20 healthy male volunteers aged 39 +/- 7 years. RESULTS: Basal AVP levels were found to be suppressed over the whole study period. In contrast, N terminal proatrial natriuretic peptide remained increased over all 280 days. No persistent alterations were found for aldosterone or angiotensin II. Sodium and potassium in plasma and urine returned to normal within a few weeks. Creatinine clearance, urea nitrogen in plasma and urine, urinary osmolality, hematocrit, and hemoglobin remained low as compared with controls over the entire study. CONCLUSIONS: Chronic alcohol abuse causes severe and persistent alterations in the hormonal regulatory systems of electrolyte and water balance. The suppressed basal secretion of AVP may reflect a dysregulation in the brain that influences the hypothalamic-pituitary-adrenal axis function, mood, memory, addiction behavior, and craving during alcohol abstinence. These findings may provide a ground for future therapeutic approaches to stable abstinence. PMID- 12766632 TI - Alcohol consumption and HIV disease progression: are they related? AB - BACKGROUND: The relationship between alcohol consumption and HIV disease progression has received limited attention in the era of highly active antiretroviral therapy (HAART). METHODS: We assessed CD4 cell count, HIV RNA levels, and alcohol consumption in the past month, defined as none, moderate, and at risk, in 349 HIV-infected people with a history of alcohol problems. We investigated the relationship between alcohol consumption and HIV disease markers CD4 cell count and HIV RNA level, stratified by HAART use, using multivariable regression. RESULTS: No significant differences in CD4 cell count or HIV RNA level were found across the categories of alcohol consumption for patients who were not receiving HAART. However, among patients who were receiving HAART, log HIV RNA levels were significantly higher in those with moderate (2.17 copies/ml) and at-risk (2.73 copies/ml) alcohol use compared with none (1.73 copies/ml; p = 0.006). CD4 cell counts in those with moderate (368 cells/microl) and at-risk (360 cells/microl) alcohol use were lower than for subjects who reported none (426 cells/microl; p = 0.07). CONCLUSION: Among patients who have a history of alcohol problems and are receiving antiretroviral treatment, alcohol consumption was associated with higher HIV RNA levels and lower CD4 counts. No comparable association was found for similar patients who were not receiving HAART. Addressing alcohol use in HIV-infected patients, especially those who are receiving HAART, may have a substantial impact on HIV disease progression. PMID- 12766633 TI - Candidate genes for alcohol dependence: a review of genetic evidence from human studies. PMID- 12766634 TI - Candidate genes for alcohol dependence: animal studies. PMID- 12766636 TI - Opioids: more to learn, improvements to be made. PMID- 12766637 TI - Individual differences in pain sensitivity: implications for treatment decisions. PMID- 12766638 TI - Preconditioning by sevoflurane decreases biochemical markers for myocardial and renal dysfunction in coronary artery bypass graft surgery: a double-blinded, placebo-controlled, multicenter study. AB - BACKGROUND: Preconditioning by volatile anesthetics is a promising therapeutic strategy to render myocardial tissue resistant to perioperative ischemia. It was hypothesized that sevoflurane preconditioning would decrease postoperative release of brain natriuretic peptide, a biochemical marker for myocardial dysfunction. In addition, several variables associated with the protective effects of preconditioning were evaluated. METHODS: Seventy-two patients scheduled for coronary artery bypass graft surgery under cardioplegic arrest were randomly assigned to preconditioning during the first 10 min of complete cardiopulmonary bypass with either placebo (oxygen-air mixture only) or sevoflurane 4 vol% (2 minimum alveolar concentration). No other volatile anesthetics were administered at any time during the study. Treatment was strictly blinded to anesthesiologists, perfusionists, and surgeons. Biochemical markers of myocardial dysfunction and injury (brain natriuretic peptide, creatine kinase-MB activity, and cardiac troponin T), and renal dysfunction (cystatin C) were determined. Results of Holter electrocardiography were recorded perioperatively. Translocation of protein kinase C was assessed by immunohistochemical analysis of atrial samples. RESULTS: Sevoflurane preconditioning significantly decreased postoperative release of brain natriuretic peptide, a sensitive biochemical marker of myocardial contractile dysfunction. Pronounced protein kinase C delta and epsilon translocation was observed in sevoflurane-preconditioned myocardium. In addition, postoperative plasma cystatin C concentrations increased significantly less in sevoflurane preconditioned patients. No differences between groups were found for perioperative ST-segment changes, arrhythmias, or creatine kinase-MB and cardiac troponin T release. CONCLUSIONS: Sevoflurane preconditioning preserves myocardial and renal function as assessed by biochemical markers in patients undergoing coronary artery bypass graft surgery under cardioplegic arrest. This study demonstrated for the first time translocation of protein kinase C isoforms delta and epsilon in human myocardium in response to sevoflurane. PMID- 12766639 TI - Auricular acupressure as a treatment for anxiety in prehospital transport settings. AB - BACKGROUND: Auricular acupuncture at the relaxation point has been previously shown to be an effective treatment for anxiety in the preoperative setting. The purpose of this prospective, randomized, blinded study was to determine whether auricular acupressure can reduce stress and anxiety during ambulance transport. METHODS: Patients who required ambulance transport secondary to medical conditions were randomized to receive auricular acupressure at the relaxation point (n = 17) or at a sham point (n = 19). A visual analog scale was used to assess state anxiety as well as patient anticipation of hospital medical treatment (estimated waiting period for treatment, anticipated pain during treatment, attitude toward the physicians, and treatment outcomes). These variables were assessed at baseline and on arrival to the hospital. RESULTS: Patients in the relaxation group reported significantly less anxiety than patients in the sham group on arrival to the hospital (visual analog scale mean +/- SD: 37.6 +/- 20.6 to 12.4 +/- 7.8 mm vs. 42.5 +/- 29.9 to 46.7 +/- 25.9 mm, respectively; P = 0.002). Similarly, patient perception of pain during treatment (mean visual analog scale +/- SD: 32.7 +/- 27.7 to 14.5 +/- 8.1 mm vs. 17.2 +/- 26.1 to 28.8 +/- 21.9 mm, respectively; P = 0.006) and treatment outcomes of their illnesses (mean visual analog scale +/- SD: 46.7 +/- 29.4 to 19.1 +/- 10.4 mm vs. 35.0 +/- 25.7 to 31.5 +/- 20.5 mm, respectively; P = 0.014) were significantly more positive in the relaxation group than in the sham group. No differences were found in the other variables assessed. CONCLUSION: It was concluded that auricular acupressure is an effective treatment for anxiety in prehospital emergency settings. PMID- 12766640 TI - Accelerometry of adductor pollicis muscle predicts recovery of respiratory function from neuromuscular blockade. AB - BACKGROUND: Residual paralysis increases the risk of pulmonary complications but is difficult to detect. To test the hypothesis that accelerometry predicts effects of residual paralysis on pulmonary and upper airway function, the authors related tests of pulmonary and pharyngeal function to accelerometry of adductor pollicis muscle in 12 partially paralyzed volunteers. METHODS: Rocuronium (0.01 mg/kg + 2-10 microg x kg-1 x min-1) was administered to maintain train-of-four (TOF) ratios (assessed every 15 s) of approximately 0.5 and 0.8 over a period of more than 5 min. The authors evaluated pharyngeal and facial muscle functions during steady state relaxation and performed spirometric measurements every 5 min until recovery. Upper airway obstruction was defined as a mean ratio of expiratory and inspiratory flow at 50% of vital capacity of greater than 1. The TOF ratio associated with "acceptable" pulmonary recovery (forced vital capacity and forced inspiratory volume in 1 s of > or =90% of baseline) was calculated using a linear regression model. RESULTS: At peak blockade (TOF ratio 0.5 +/- 0.16), forced inspiratory flow was impaired (53 +/- 19%) to a greater degree than forced expiratory flow (75 +/- 20%) with a mean ratio of expiratory and inspiratory flow at 50% of vital capacity of 1.18 +/- 0.6. Upper airway obstruction, observed in 8 of 12 volunteers, paralleled an impaired ability to swallow reported by 10 of 12 volunteers. In contrast, all volunteers except one could sustain a head lift for more than 5 s. The authors calculated that a mean TOF ratio of 0.56 (95% confidence interval, 0.22-0.71) predicts "acceptable" recovery of forced vital capacity, whereas forced inspiratory volume in 1 s was impaired until a TOF ratio of 0.95 (0.82-1.18) was reached. A 100% recovery of TOF ratio predicts an acceptable recovery of forced vital capacity, forced inspiratory volume in 1 s, and mean ratio of expiratory and inspiratory flow at 50% of vital capacity in 93%, 73%, and 88% of measurements (calculated negative predictive values), respectively. CONCLUSION: Impaired inspiratory flow and upper airway obstruction frequently occur during minimal neuromuscular blockade (TOF ratio 0.8), and extubation may put the patient at risk. Although a TOF ratio of unity predicts a high probability of adequate recovery from neuromuscular blockade, respiratory function can still be impaired. PMID- 12766641 TI - Terlipressin versus norepinephrine to correct refractory arterial hypotension after general anesthesia in patients chronically treated with renin-angiotensin system inhibitors. AB - BACKGROUND: Terlipressin, a precursor that is metabolized to lysine-vasopressin, has been proposed as a drug for treatment of intraoperative arterial hypotension refractory to ephedrine in patients who have received long-term treatment with renin-angiotensin system inhibitors. The authors compared the effectiveness of terlipressin and norepinephrine to correct hypotension in these patients. METHODS: Among 42 patients scheduled for elective carotid endarterectomy, 20 had arterial hypotension following general anesthesia that was refractory to ephedrine. These patients were the basis of the study. After randomization, they received either 1 mg intravenous terlipressin (n = 10) or norepinephrine infusion (n = 10). Beat-by-beat recordings of systolic arterial blood pressure and heart rate were stored on a computer. The intraoperative maximum and minimum values of blood pressure and heart rate, and the time spent with systolic arterial blood pressure below 90 mmHg and above 160 mmHg, were used as indices of hemodynamic stability. Data are expressed as median (95% confidence interval). RESULTS: Terlipressin and norepinephrine corrected arterial hypotension in all cases. However, time spent with systolic arterial blood pressure below 90 mmHg was less in the terlipressin group (0 s [0-120 s] vs. 510 s [120-1011 s]; P < 0.001). Nonresponse to treatment (defined as three boluses of terlipressin or three changes in norepinephrine infusion) occurred in zero and eight cases (P < 0.05), respectively. CONCLUSIONS: In patients who received long-term treatment with renin-angiotensin system inhibitors, intraoperative refractory arterial hypotension was corrected with both terlipressin and norepinephrine. However, terlipressin was more rapidly effective for maintaining normal systolic arterial blood pressure during general anesthesia. PMID- 12766642 TI - Simulation study of rested versus sleep-deprived anesthesiologists. AB - BACKGROUND: Sleep deprivation causes physiologic and subjective sleepiness. Studies of fatigue effects on anesthesiologist performance have given equivocal results. The authors used a realistic simulation environment to study the effects of sleep deprivation on psychomotor and clinical performance, subjective and objective sleepiness, and mood. METHODS: Twelve anesthesia residents performed a 4-h anesthetic on a simulated patient the morning after two conditions of prior sleep: sleep-extended (EXT), in which subjects were allowed to arrive at work at 10:00 AM for 4 consecutive days, thus allowing an increase in nocturnal sleep time, and total sleep deprivation (DEP), in which subjects were awake at least 25 h. Psychomotor testing was performed at specified periods throughout the night in the DEP condition and at matched times during the simulation session in both conditions. Three types of vigilance probes were presented to subjects at random intervals as well as two clinical events. Task analysis and scoring of alertness were performed retrospectively from videotape. RESULTS: In the EXT condition, subjects increased their sleep by more than 2 h from baseline (P = 0.0001). Psychomotor tests revealed progressive impairment of alertness, mood, and performance in the DEP condition over the course of the night and when compared with EXT during the experimental day. DEP subjects showed longer response latency to vigilance probes, although this was statistically significant for only one probe type. Task analysis showed no difference between conditions except that subjects "slept" more in the DEP condition. There was no significant difference in the cases' clinical management between sleep conditions. Subjects in the DEP condition had lower alertness scores (P = 0.02), and subjects in the EXT condition showed little video evidence of sleepiness. CONCLUSIONS: Psychomotor performance and mood were impaired while subjective sleepiness and sleepy behaviors increased during simulated patient care in the DEP condition. Clinical performance between conditions was similar. PMID- 12766643 TI - Effects of antidepressants on function and viability of human neutrophils. AB - BACKGROUND: Antidepressants are frequently used in chronic pain therapy and are under investigation as long-acting local anesthetics. Because of the structural similarities between antidepressants and local anesthetics, the authors hypothesized that these compounds act similarly, and they investigated the effects of nortriptyline, amitriptyline, imipramine, and fluoxetine on priming and activation of human polymorphonuclear neutrophils (hPMNs). METHODS: Effects of 30-, 120-, and 240-min preincubation with nortriptyline (10(-7)-10(-4) M), amitriptyline (10(-6)-10(-3) M), imipramine (10(-6)-10(-3) M), or fluoxetine (10( 7)-10(-4) M) on O(2)- generation of platelet activating factor-primed (10-6 M) and/or formyl-methionyl-leucyl-phenylalanine-activated (10(-6) M) isolated hPMNs were determined. All data are reported as mean +/- SD (statistics: t test, P < 0.05). RESULTS: Brief incubation in low concentrations of nortriptyline, amitriptyline, or fluoxetine (all at 10(-5) M) did inhibit priming but not activation of hPMNs. Imipramine (10(-5) M) affected neither priming nor activation. Prolonged incubation in lower concentrations of all antidepressants influenced neither priming nor activation. However, at higher concentrations, all four compounds exerted cytotoxic effects: virtually all hPMNs were killed by amitriptyline and imipramine (both at 10(-3) M) or nortriptyline and fluoxetine (both at 10(-4) M). CONCLUSION: Antidepressants, in low concentrations, inhibited priming but not activation of hPMNs. However, at concentrations similar to those attained after local injection, and in marked contrast to local anesthetics, antidepressants are profoundly toxic to hPMNs. PMID- 12766645 TI - Temporal summation governs part of the minimum alveolar concentration of isoflurane anesthesia. AB - BACKGROUND: General anesthesia may delay the onset of movement in response to noxious stimulation. The authors hypothesized that the production of immobility could involve depression of time-related processes involved in the generation of movement. METHODS: The delays (latencies) between onset of tail clamp (n = 16) or 50-Hz continuous electrical stimulation (n = 8) and movement were measured in rats equilibrated at 0.1-0.2% increasing steps of isoflurane. In other rats (n = 8), the isoflurane concentrations just permitting and preventing movement (crossover concentrations) in response to trains of 0.5-ms 50-V square-wave pulses of interstimulus intervals of 10, 3, 1, 0.3, or 0.1 s during the step increases were measured. These measures were again made during administration of intravenous MK801, an N-methyl-D-aspartate receptor antagonist that can block temporal summation (n = 6). Temporal summation refers to the cumulative effect of repeated stimuli. Crossover concentrations to 10- and 0.1-s interstimulus interval pulses ranging in voltage from 0.25-50 V were also measured (n = 4). RESULTS: The increase in concentrations from 0.6 to nearly 1.0 minimum alveolar concentration progressively increased latency from less than 1 s to 58 s. Shortening the interstimulus interval (50 V) pulses from 10 to 0.1 s progressively increased crossover concentrations from 0.6 to 1.0 minimum alveolar concentration. In contrast, during MK801 administration shortening interstimulus intervals did not change crossover concentrations, producing a flat response to change in the interstimulus interval. Increasing the voltage of interstimulus interval pulses increased the crossover concentrations but did not change the response to change in interstimulus intervals for pulses greater than 1 V. CONCLUSIONS: Increasing the duration or frequency (interstimulus interval) of stimulation increases the concentration of isoflurane required to suppress movement by a 0.4 minimum alveolar concentration MK801 blocks this effect, a finding consistent with temporal summation (which requires intact N-methyl-D aspartate receptor activity) at concentrations of up to 1 minimum alveolar concentration isoflurane. PMID- 12766644 TI - Propofol increases phosphorylation of troponin I and myosin light chain 2 via protein kinase C activation in cardiomyocytes. AB - BACKGROUND: Troponin I (TnI) and myosin light chain 2 (MLC2) are important myofibrillar proteins involved in the regulation of myofilament calcium (Ca2+) sensitivity and cardiac inotropy. The objectives of this study were to determine the role of protein kinase C (PKC) in mediating propofol-induced changes in actomyosin adenosine triphosphatase activity in cardiac myofibrils and to examine the extent to which propofol alters the phosphorylation of TnI and MLC2 in cardiomyocytes. METHODS: Freshly isolated adult rat ventricular myocytes were used for the study. Cardiac myofibrils were extracted for assessment of actomyosin adenosine triphosphatase activity and phosphorylation of TnI and MLC2. Western blot analysis for PKC-alpha was performed on cardiomyocyte subcellular fractions. Simultaneous measurement of intracellular free Ca2+ concentration ([Ca2+](i)) and myocyte shortening was assessed using fura-2 and video edge detection, respectively. RESULTS: Propofol (30 microM) reduced the Ca2+ concentration required for activation of actomyosin adenosine triphosphatase activity, and this effect was abolished by bisindolylmaleimide I. In addition, propofol stimulated dose-dependent phosphorylation of TnI and MLC2. PKC activation with phorbol myristic acetate also stimulated an increase in phosphorylation of TnI and MLC2. The actions of propofol and phorbol myristic acetate together on phosphorylation of TnI and MLC2 were not additive. PKC inhibition with bisindolylmaleimide I attenuated phorbol myristic acetate- and propofol-induced phosphorylation of TnI and MLC2. Propofol stimulated translocation of PKC-alpha from cytosolic to membrane fraction. Propofol caused a shift in the extracellular Ca2+-shortening relationship, and this effect was abolished by bisindolylmaleimide I. CONCLUSIONS: These results suggest that propofol increases myofilament Ca2+ sensitivity via a PKC-dependent pathway involving the phosphorylation of MLC2. PMID- 12766646 TI - Cyclooxygenase-2-dependent superoxide generation contributes to age-dependent impairment of G protein-mediated cerebrovasodilation. AB - BACKGROUND: Previous studies have observed that activation of cyclooxygenase-2 contributes to generation of superoxide anion after fluid percussion brain injury (FPI). This study was designed to characterize the effects of FPI on the vascular activity of two activators of a pertussis toxin-sensitive G protein, mastoparan and mastoparan-7, and the role of cyclooxygenase-2-dependent superoxide anion generation in such effects as a function of age. METHODS: Lateral FPI was induced in anesthetized newborn (1-5-day-old) and juvenile (3-4-week-old) pigs equipped with a closed cranial window. RESULTS: Mastoparan (10(-8), 10(-6) M) elicited pial artery dilation that was blunted more in newborn versus juvenile pigs (9 +/- 1 and 16 +/- 1 vs. 3 +/- 1 and 5 +/- 1%, newborn; 9 +/- 1 and 15 +/- 1 vs. 6 +/- 1 and 9 +/- 1%, juveniles). Similar results were observed for mastoparan-7 but the inactive analog mastoparan-17 had no effect on pial artery diameter. Indomethacin (a cyclooxygenase-1 and cyclooxygenase-2 inhibitor), NS398 (a cyclooxygenase-2 inhibitor), and polyethylene glycol superoxide dismutase and catalase (free radical scavengers) partially restored impaired mastoparan dilation after FPI in the newborn in a roughly equivalent manner but not in the juvenile (3 +/- 1 and 5 +/- 1 vs. 8 +/- 1 and 13 +/- 1% newborn, 6 +/- 1 and 9 +/ 1 vs. 7 +/- 1 and 10 +/- 1% juvenile for NS398 pretreatment). CONCLUSIONS: These data show that G protein activation elicits cerebrovasodilation that is blunted following FPI in an age-dependent manner, and suggest that cyclooxygenase-2 dependent superoxide anion generation contributes to G protein activation-induced dilator impairment after the insult in an age-dependent manner. PMID- 12766647 TI - N-acetylcysteine restores isoflurane-induced preconditioning against myocardial infarction during hyperglycemia. AB - BACKGROUND: Hyperglycemia generates reactive oxygen species and prevents isoflurane-induced preconditioning. The authors tested the hypothesis that scavenging reactive oxygen species with N-acetylcysteine will restore protection against myocardial infarction produced by isoflurane in vivo. METHODS: Barbiturate-anesthetized dogs (n = 45) were instrumented for measurement of systemic hemodynamics. Myocardial infarct size and coronary collateral blood flow were measured with triphenyltetrazolium staining and radioactive microspheres, respectively. All dogs were subjected to a 60-min left anterior descending coronary artery occlusion followed by 3 h of reperfusion. Dogs were randomly assigned to receive an infusion of 0.9% saline or 15% dextrose in water to increase blood glucose concentrations to 600 mg/dl (hyperglycemia) in the absence or presence of isoflurane (1.0 minimum alveolar concentration) with or without pretreatment with N-acetylcysteine (150 mg/kg i.v.) in six experimental groups. Isoflurane was discontinued, and blood glucose concentrations were allowed to return to baseline values before left anterior descending coronary artery occlusion. RESULTS: Myocardial infarct size was 27 +/- 2% (n = 8) of the left ventricular area at risk in control experiments. Isoflurane significantly (P < 0.05) decreased infarct size (13 +/- 2%; n = 7). Hyperglycemia alone did not alter infarct size (29 +/- 3%; n = 7) but abolished the protective effect of isoflurane (25 +/- 2%; n = 8). N-Acetylcysteine alone did not affect infarct size (28 +/- 2%; n = 8) but restored isoflurane-induced cardioprotection during hyperglycemia (10 +/- 1%; n = 7). CONCLUSIONS: Acute hyperglycemia abolishes reductions in myocardial infarct size produced by isoflurane, but N acetylcysteine restores these beneficial effects. The results suggest that excessive quantities of reactive oxygen species generated during hyperglycemia impair isoflurane-induced preconditioning in dogs. PMID- 12766648 TI - Perflubron emulsion in prolonged hemorrhagic shock: influence on hepatocellular energy metabolism and oxygen-dependent gene expression. AB - BACKGROUND: Liver dysfunction as a result of impaired oxygen availability frequently occurs following hemorrhage and contributes to delayed mortality. Artificial oxygen carriers may improve oxygen supply to vital organs while avoiding the need for allogeneic transfusion. METHODS: Rats were subjected to hemorrhagic hypotension (mean arterial pressure = 35-40 mmHg for 120 min) and were subsequently resuscitated with (1) stored whole rat blood, (2) pentastarch, or (3) pentastarch combined with perflubron emulsion (PFE; 2.7 or 5.4 g/kg body weight), a second-generation artificial oxygen carrier. Recovery of liver adenosine triphosphate, hepatocellular injury, and expression of glutamine synthetase 1, a gene that is induced by exposure of hepatocytes to low partial pressure of oxygen, were studied at 4 h of resuscitation. RESULTS: Stored whole blood or pentastarch failed to restore liver adenosine triphosphate concentrations after prolonged shock as compared to sham controls and resulted in increased gene expression of glutamine synthetase 1. Addition of 2.7 g PFE/kg restored liver adenosine triphosphate to control, whereas 5.4 g PFE/kg resulted in adenosine triphosphate concentrations significantly above control. Improved hepatocellular oxygen supply was also confirmed by restoration of the physiologic expression pattern of glutamine synthetase 1. Serum enzyme concentrations were highest after resuscitation with stored blood, whereas addition of PFE failed to further decrease enzyme concentrations as compared to pentastarch alone. CONCLUSIONS: Resuscitation with PFE is superior to stored blood or asanguineous resuscitation with respect to restoration of hepatocellular energy metabolism. The improved hepatocellular oxygen availability is reflected in normalization of oxygen-dependent gene expression. However, improved oxygen availability failed to affect early hepatocellular injury. PMID- 12766649 TI - Fetal plasma concentrations after intraamniotic sufentanil in chronically instrumented pregnant sheep. AB - BACKGROUND: Rapid progress is being made in fetal surgery. Because the fetus is capable of pain perception after the 26th week of gestation, adequate postoperative fetal pain management is essential. The preferred approach would provide fetal analgesia without affecting the mother. Intraamniotically administered sufentanil may be an interesting option if it achieves therapeutic plasma concentrations (PCs) in the fetus but not the mother. METHODS: After approval of the study, 25 or 50 microg sufentanil was administered intraamniotically in 10 chronically instrumented pregnant ewes. Maternal and fetal vital signs, arterial blood gases, and uterine blood flow were recorded over 120 min. Sufentanil PCs were determined before and 1, 3, 5, 10, 15, 30, 45, 60, 90, and 120 min after injection. Statistical analysis was performed using one or two-way analysis of variance followed by Dunnett or Tukey test, as appropriate (P < 0.05; data presented as median [95% confidence interval]). RESULTS: After 25 microg sufentanil, fetal PC stabilized at 134 +/- 89 pg/ml (after 10 min), and maternal PCs stabilized at 44 +/- 11 pg/ml (after 15 min). After 50 microg sufentanil, fetal PCs stabilized at 134 +/- 35 pg/ml (after 15 min), and maternal PCs reached 80 +/- 25 pg/ml (at 30 min). Injection of 25 microg sufentanil intraamniotically did not affect maternal or fetal hemodynamics, uterine blood flow, or arterial blood gases. Fetal heart rate increased after administration of 50 microg sufentanil (maximum change at 10 min: +16 +/- 12%). CONCLUSION: The sheep fetus absorbs sufentanil after intraamniotic instillation. Significantly greater PCs were obtained in the fetal lamb as compared with the ewe. This suggests that investigation of intraamniotic opioids for fetal analgesia might be worthwhile. PMID- 12766650 TI - Olprinone, a phosphodiesterase III inhibitor, reduces gut mucosal injury and portal endotoxin level during acute hypoxia in rabbits. AB - BACKGROUND: Preservation of gut integrity has become a therapeutic goal to obviate bacterial translocation in the critically ill. The authors examined whether olprinone, a phosphodiesterase III inhibitor, protected functional and structural integrity of gut mucosa against acute progressive hypoxia. METHODS: Thirty-two animals were randomly allocated to a control group (n = 12), a low dose group (0.2 microg x kg-1 x min-1 olprinone; n = 10), or a high-dose group (0.6 microg x kg-1 x min-1 olprinone; n = 10) after preparatory surgery. Ascending aortic and portal blood flow, intramural pH of the ileum, and portal endotoxin levels were measured at normoxia and through three stages of progressive hypoxia (fraction of inspired oxygen = 0.17, 0.13, and 0.10). RESULTS: At normoxia, ascending aortic flow in the high-dose group was approximately 20% higher than in the control and low-dose groups. During progressive hypoxia, both ascending aortic and portal flow in the control group were depressed, whereas olprinone infusion attenuated such alterations and redistributed blood to the splanchnic area in a dose-dependent manner. On the contrary, the reduction of intramural pH of the ileum and the elevation of portal endotoxin levels observed in the control group were significantly minimized in both the low- and high-dose groups to a similar extent during acute hypoxia. Histopathologic alterations of gut mucosa observed in the control group were minimized by olprinone infusion dose-independently, accompanied by reduction of mortality rate of the animals. CONCLUSIONS: Olprinone slows progression of intestinal mucosal acidosis and gut barrier dysfunction, concurrently with preservation of microscopic structures, through both flow-dependent and independent mechanisms under acute hypoxia. Such properties of olprinone may serve to protect the host under insult. PMID- 12766651 TI - Relationships between measurement of pain using visual analog score and morphine requirements during postoperative intravenous morphine titration. AB - BACKGROUND: Although intravenous morphine titration is widely used to obtain rapid and complete postoperative pain relief, the relationship between measurement of pain and morphine requirements varies, and the evolution of pain during titration is poorly understood. METHODS: Intravenous morphine titration was administered as a bolus of 2 (body weight < or = 60 kg) or 3 mg (body weight > 60 kg) during the immediate postoperative period in the PACU. The interval between each bolus was 5 min. The visual analog scale (VAS) score threshold required to administer morphine was 30, and pain relief was defined as a VAS score of 30 or less. RESULTS: Data from 3,045 patients were analyzed. The mean initial VAS score was 73 +/- 19 (mean +/- SD), and the mean morphine dose required to obtain pain relief was 0.17 +/- 0.10 mg/kg, i.e., a median of four boluses (range, 1-20). When patients were grouped according to several classes of initial VAS score (31-39, 40-49, 50-59, 60-69, 70-79, 80-89, 90-100), it seemed that the relationship between VAS score and morphine requirements was a sigmoid curve. A VAS score of 70 or greater predicted the need for a high (>0.15 mg/kg) morphine dose (sensitivity, 0.77; specificity, 0.54). During the pain relief process, the relationship between VAS score and time was depicted by a sigmoid curve. CONCLUSION: A VAS score of 70 or greater should be considered indicative of severe pain. The relationship between the initial VAS score and morphine requirements is not linear, and the evolution of the VAS score during the pain relief process is described by a sigmoid curve. PMID- 12766652 TI - Postcesarean section pain prediction by preoperative experimental pain assessment. AB - BACKGROUND: Postcesarean section pain is a common cause of acute pain in obstetrics, yet pain relief and patient satisfaction are still inadequate in many cases. The present study was conducted to determine whether preoperative assessment of experimental pain perception by quantitative sensory tests could predict the level of postcesarean section pain. METHODS: Fifty-eight women who were scheduled for elective cesarean section were enrolled in the study. Heat pain threshold and magnitude estimation of suprathreshold pain stimuli at 44 degrees-48 degrees C were assessed for both algosity (the sensory dimension of pain intensity) and unpleasantness 1 or 2 days before surgery. The day after the operation, the women reported the level of pain at the surgical wound on a visual analog scale at rest and during activity. Regression analysis was performed to evaluate the usefulness of preoperative scores in predicting postcesarean section pain. RESULTS: Postoperative visual analog scale scores at rest and during activity significantly correlated with preoperative suprathreshold pain scores at 44 degrees-48 degrees C (r = 0.31-0.58 for algosity and r = 0.33-0.74 for unpleasantness). The stimulus of 48 degrees C was found to be the best predictor of postoperative pain for both conditions (r = 0.434-0.527; P < 0.01). In contrast to suprathreshold pain stimuli, pain threshold was not correlated with postoperative pain. CONCLUSIONS: The results show that a simple and quick preoperative test is useful in identifying those women who will experience greater pain after a cesarean section. This test may be suggested for caregivers to tailor the postoperative treatment to specific patient needs and to improve postoperative outcome and patient satisfaction. PMID- 12766653 TI - Bilateral intravenous regional anesthesia: a new method to test additives to local anesthetic solutions. AB - BACKGROUND: Ketorolac, when added to lidocaine, has been shown to reduce early tourniquet pain during intravenous regional anesthesia (i.v.RA) in patients. Although the effectiveness of ropivacaine 0.2% for i.v.RA is equal to that of lidocaine 0.5% but significantly reduces central nervous system side effects after release of the tourniquet, it provides no advantage with regard to tourniquet tolerance times. Simultaneous bilateral i.v.RA with ropivacaine 0.2% was used to test the hypothesis that ketorolac modifies tourniquet tolerance and to test whether drug combinations can be evaluated in one study session. METHODS: Ten healthy, unsedated volunteers received 30 ml of ropivacaine 0.2% in each upper arm with 2 ml of normal saline in one arm and 30 mg of ketorolac in the contralateral arm for i.v.RA. Both proximal tourniquets remained inflated for 30 min, followed by inflation of the distal tourniquets and release of the proximal ones. Verbal numeric scores for tourniquet pain were recorded for both extremities. Central nervous system side effects were graded after release of each distal tourniquet. RESULTS: There was no difference between the two upper extremities with regard to surgical anesthesia and tourniquet tolerance. Total tourniquet tolerance was a median of 58.5 min (range, 45-90 min) and 60.5 min (39 79 min) in the normal saline and ketorolac groups, respectively. After release of the distal tourniquets, 5 of 10 volunteers experienced mild dizziness. CONCLUSIONS: The addition of ketorolac to ropivacaine does not improve tourniquet tolerance. Minimal central nervous system side effects after tourniquet release suggest that a total of 60 ml ropivacaine 0.2% for bilateral i.v.RA is a useful model for comparison of i.v.RA drug combinations. PMID- 12766654 TI - A multicenter, randomized, controlled trial comparing bupivacaine with ropivacaine for labor analgesia. AB - BACKGROUND: A meta-analysis of studies comparing high doses of bupivacaine with ropivacaine for labor pain found a higher incidence of forceps deliveries, motor block, and poorer neonatal outcome with bupivacaine. The purpose of this study was to determine if there is a difference in these outcomes when a low concentration of patient-controlled epidural bupivacaine combined with fentanyl is compared with ropivacaine combined with fentanyl. METHODS: This was a multicenter, randomized, controlled trial, including term, nulliparous women undergoing induction of labor. For the initiation of analgesia, patients were randomized to receive either 15 ml bupivacaine, 0.1%, or 15 ml ropivacaine, 0.1%, each with 5 microg/ml fentanyl. Analgesia was maintained with patient-controlled analgesia with either local anesthetic, 0.08%, with 2 microg/ml fentanyl. The primary outcome was the incidence of operative delivery. We also examined other obstetric, neonatal, and analgesic outcomes. RESULTS: There was no difference in the incidence of operative delivery between the two groups (148 of 276 bupivacaine recipients vs. 135 of 279 ropivacaine recipients; P = 0.25) or any obstetric or neonatal outcome. The incidence of motor block was significantly increased in the bupivacaine group compared with the ropivacaine group at 6 h (47 of 93 vs. 29 of 93, respectively; P = 0.006) and 10 h (29 of 47 vs. 16 of 41, respectively; P = 0.03) after injection. Satisfaction with mobility was higher with ropivacaine than with bupivacaine (mean +/- SD: 76 +/- 23 vs. 72 +/- 23, respectively; P = 0.013). Satisfaction for analgesia at delivery was higher for bupivacaine than for ropivacaine (mean +/- SD: 71 +/- 25 vs. 66 +/- 26, respectively; P = 0.037). CONCLUSIONS: There was no difference in the incidence of operative delivery or neonatal outcome among nulliparous patients who received low concentrations of bupivacaine or ropivacaine for labor analgesia. PMID- 12766655 TI - Comparison of the parasacral approach and the posterior approach, with single- and double-injection techniques, to block the sciatic nerve. AB - BACKGROUND: The purpose of this study was to compare parasacral and Winnie's single- or double-injection approaches for sciatic nerve block. METHODS: One hundred fifty adults scheduled to undergo lower limb surgery were randomized to receive on the sciatic nerve 20 ml ropivacaine, 0.75%: single bolus for parasacral and Winnie's single injection. For Winnie's double injection, the peroneal and tibial nerves received separately 10 ml plus 10 ml. Blocks were performed with the use of nerve stimulator (intensity < 0.5 mA, 1 Hz). For the parasacral method, a line was drawn between the posterior superior iliac spine and the ischial tuberosity; needle entry was at 6 cm inferior to the posterior superior iliac spine. RESULTS: The groups were similar. Time to perform the block was 2 (1-5) min for the parasacral method, with no difference from Winnie's single injection (3 [1-10] min), but was shorter with double injection (5.5 [2 15] min) (P = 0.0001). Onset of sensory block was similar in the parasacral (25 [7.5-50] min) and Winnie single-injection groups (25 [5-50] min) but significantly longer in the double-injection group (15 [5-50] min). Success rates for complete block were similar in the parasacral (66%) and Winnie's double injection groups (68%) after 30 min but higher in the Winnie's single-injection group for tibial sensory and motor block (48%) (P < 0.017). CONCLUSION: Time to perform a parasacral block was short, and the parasacral approach had a high success rate and a short onset time. Therefore, this block might be a useful alternative to Winnie's modification for sciatic nerve block. PMID- 12766656 TI - Piriformis syndrome: anatomic considerations, a new injection technique, and a review of the literature. AB - BACKGROUND: Piriformis syndrome can be caused by anatomic abnormalities. The treatments of piriformis syndrome include the injection of steroid into the piriformis muscle and near the area of the sciatic nerve. These techniques use either fluoroscopy and muscle electromyography to identify the piriformis muscle or a nerve stimulator to stimulate the sciatic nerve. METHODS: The authors performed a cadaver study and noted anatomic variations of the piriformis muscle and sciatic nerve. To standardize their technique of injection, they also noted the distance from the lower border of the sacroiliac joint (SIJ) to the sciatic nerve. They retrospectively reviewed the charts of 19 patients who had received piriformis muscle injections, noting the site of needle insertion in terms of the distance from the lower border of the SIJ and the depth of needle insertion at which the motor response of the foot was elicited. The authors tabulated the response of the patients to the injection, any associated diagnoses, and previous treatments that these patients had before the injection. Finally, they reviewed the literature on piriformis syndrome, a rare cause of buttock pain and sciatica. RESULTS: In the cadavers, the distance from the lower border of the SIJ to the sciatic nerve was 2.9 +/- 0.6 (1.8-3.7) cm laterally and 0.7 +/- 0.7 (0.0-2.5) cm caudally. In 65 specimens, the sciatic nerve passed anterior and inferior to the piriformis. In one specimen, the muscle was bipartite and the two components of the sciatic nerve were separate, with the tibial nerve passing below the piriformis and the peroneal nerve passing between the two components of the muscle. In the patients who received the injections, the site of needle insertion was 1.5 +/- 0.8 (0.4-3.0) cm lateral and 1.2 +/- 0.6 (0.5-2.0) cm caudal to the lower border of the SIJ as seen on fluoroscopy. The needle was inserted at a depth of 9.2 +/- 1.5 (7.5-13.0) cm to stimulate the sciatic nerve. Patients had comorbid etiologies including herniated disc, failed back surgery syndrome, spinal stenosis, facet syndrome, SIJ dysfunction, and complex regional pain syndrome. Sixteen of the 19 patients responded to the injection, their improvements ranged from a few hours to 3 months. CONCLUSIONS: Anatomic abnormalities causing piriformis syndrome are rare. The technique used in the current study was successful in injecting the medications near the area of the sciatic nerve and into the piriformis muscle. PMID- 12766657 TI - Postoperative morphine use and hyperalgesia are reduced by preoperative but not intraoperative epidural analgesia: implications for preemptive analgesia and the prevention of central sensitization. AB - BACKGROUND: The aim of this study was to evaluate the postoperative morphine sparing effects and reduction in pain and secondary mechanical hyperalgesia after preincisional or postincisional epidural administration of a local anesthetic and an opioid compared with a sham epidural control. METHODS: Patients undergoing major gynecologic surgery by laparotomy were randomly assigned to three groups and studied in a double-blinded manner. Group 1 received epidural lidocaine and fentanyl before incision and epidural saline 40 min after incision. Group 2 received epidural saline before incision and epidural lidocaine and fentanyl 40 min after incision. Group 3 received a sham epidural control (with saline injected into a catheter taped to the back) before and 40 min after incision. All patients underwent surgery with general anesthesia. RESULTS: One hundred forty one patients completed the study (group 1, n = 45; group 2, n = 49; group 3, n = 47). Cumulative patient-controlled analgesia morphine consumption at 48 h was significantly lower (P = 0.04) in group 1 (89.8 +/- 43.3 mg) than group 3 (112.5 +/- 71.5 mg) but not group 2 (95.4 +/- 60.2 mg), although the hourly rate of morphine consumption between 24 and 48 h after surgery was significantly lower (P < 0.0009) in group 1 (1.25 +/- 0.02 mg/h) than group 2 (1.41 +/- 0.02 mg/h). Twenty-four hours after surgery, the visual analog scale pain score on movement was significantly less intense (P = 0.005) in group 1 (4.9 +/- 2.2 cm) than group 3 (6.0 +/- 2.6 cm) but not group 2 (5.3 +/- 2.5 cm), and the von Frey pain threshold near the wound was significantly higher (P = 0.03) in group 1 (6.4 +/- 0.6 log mg) than in group 3 (6.1 +/- 0.8 log mg) but not group 2 (6.2 +/- 0.7 log mg). CONCLUSIONS: Preincisional administration of epidural lidocaine and fentanyl was associated with a significantly lower rate of morphine use, lower cumulative morphine consumption, and reduced hyperalgesia compared with a sham epidural condition. These results highlight the importance of including a standard treatment control group to avoid the problems of interpretation that arise when two-group studies of preemptive analgesia (preincisional vs. postsurgery) fail to find the anticipated effects. PMID- 12766658 TI - Intraspinal adenosine induces spinal cord norepinephrine release in spinal nerve ligated rats but not in normal or sham controls. AB - BACKGROUND: Intrathecal adenosine is antinociceptive under conditions of central sensitization, but not in response to acute stimuli in normals. The reasons for this selective circumstance of action remain unclear, but some evidence links adenosine's antinociceptive effects to release of norepinephrine by terminals in the spinal cord. The purpose of this study was to test whether spinal adenosine induces norepinephrine release selectively in settings of hypersensitivity. METHODS: Rats randomly assigned to spinal nerve ligation, sham operation, or no operation were anesthetized. A microdialysis fiber was implanted in the spinal cord dorsal horn at the L5-L6 level and perfused with artificial cerebrospinal fluid. After washout and a baseline sample period, adenosine at various concentrations was infused through the fiber for 150 min, and samples were collected every 15 min. RESULTS: In ligated, but not in sham or normal animals, adenosine perfusion increased norepinephrine in spinal cord microdialysates in a concentration-dependent manner. The effects of adenosine plateaued after 75 min and remained stable until the end of the experiment. Intravenous injection of selective adenosine A1 and A2 receptor antagonists revealed that adenosine's effect on spinal norepinephrine release was A1 receptor mediated. CONCLUSIONS: This is the first study to provide direct evidence that adenosine is able to release norepinephrine in spinal cord dorsal horns in living animals. However, this effect was only seen in animals after spinal nerve ligation. These data are consistent with behavioral studies demonstrating that adenosine's antinociceptive effects in rats after spinal nerve ligation is totally dependent on intact spinal noradrenergic terminals and can be blocked by spinal alpha 2-adrenergic receptor antagonists. PMID- 12766659 TI - Intrathecal morphine reduces the visceromotor response to acute uterine cervical distension in an estrogen-independent manner. AB - BACKGROUND: Acute uterine cervical distension (UCD) forms the basis for obstetric and some gynecologic pain. Systemic morphine inhibits the visceromotor response to UCD in rats by an action in the central nervous system, but the effect of morphine is blocked by exposure to estrogen. The purpose of the present study was to determine whether this estrogen blockade of the action of morphine reflects a spinal mechanism. METHODS: Virgin Sprague-Dawley rats received estrogen or placebo treatment for 1 week after ovariectomy. Rats were then anesthetized, and the electromyographic response in the rectus abdominis muscle to UCD was recorded in the absence and presence of cumulative dosing with intrathecal morphine. RESULTS: Estrogen treatment did not alter the stimulus- response relationship between UCD and reflex muscle contraction. Intrathecal morphine reduced the visceromotor reflex response to UCD in a dose-dependent manner that was unaffected by estrogen treatment. CONCLUSIONS: These data suggest that intrathecal morphine is effective in reducing the visceromotor response to UCD and that the reduction in efficacy of systemic morphine in this model is unlikely to reflect a reduction of the efficacy of morphine at the spinal level. These data agree with clinical studies that indicate that systemic morphine, in doses that reduce acute postoperative pain, have minimal to no effect in women in labor, yet intrathecal injection of opioids provides rapid, complete analgesia. PMID- 12766660 TI - Magnesium chloride and ruthenium red attenuate the antiallodynic effect of intrathecal gabapentin in a rat model of postoperative pain. AB - BACKGROUND: Gabapentin, a gamma-aminobutyric acid analog anticonvulsant, has been shown to possess antinociceptive effects in animal models and clinical trials. An endogenous binding site of [3H]gabapentin has been revealed to be the alpha(2)delta subunit of voltage-dependent Ca2+ channels. Magnesium chloride, ruthenium red, and spermine have been shown to modulate [3H]gabapentin binding to this binding site in vitro. In this study, the authors examined whether intrathecal magnesium chloride, ruthenium red, or spermine could affect the antiallodynic effect of intrathecal gabapentin in a rat model of postoperative pain. METHODS: Under isoflurane anesthesia, male Sprague-Dawley rats received an incision over the plantar surface of the right hind paw to produce punctate mechanical allodynia. Withdrawal thresholds to von Frey filament stimulation near the incision site were measured before incision, 2 h after incision, and every 30 min after intrathecal coadministration of gabapentin with normal saline or different doses of magnesium chloride, ruthenium red, or spermine for 2 h. RESULTS: Intrathecal gabapentin (30, 100, 200 microg) dose-dependently reduced incision-induced allodynia. Hexahydrated magnesium chloride (5, 10, 20 microg) and ruthenium red (0.2, 2, 20 ng) noncompetitively inhibited the antiallodynic effect of gabapentin. Spermine at doses not inducing motor weakness (30, 60 microg) did not affect the antiallodynic effect of gabapentin. The antiallodynic effect of intrathecal morphine (1.5 microg) was not affected by hexahydrated magnesium chloride (20 microg), ruthenium red (20 ng), or spermine (60 microg). CONCLUSIONS: These results provide behavioral evidence to support that the alpha(2)delta subunit of Ca2+ channels may be involved in the antiallodynic action of intrathecal gabapentin in the postoperative pain model. PMID- 12766661 TI - Antihyperalgesic and side effects of intrathecal clonidine and tizanidine in a rat model of neuropathic pain. AB - BACKGROUND: Although intrathecal clonidine produces pronounced analgesia, antinociceptive doses of intrathecal clonidine produce several side effects, including hypotension, bradycardia, and sedation. Intrathecal tizanidine, another alpha(2)-adrenergic agonist, has provided antinociception without producing pronounced hemodynamic changes in animal studies. However, it has been unclear whether antihyperalgesic doses of intrathecal clonidine and tizanidine produce hypotension and bradycardia in a neuropathic pain state. This study was designed to evaluate the antihyperalgesic effects and side effects of intrathecal clonidine and tizanidine in a rat model of neuropathic pain. METHODS: Male Sprague-Dawley rats were chronically implanted with lumbar intrathecal catheters, and the sciatic nerve was loosely ligated. After 21-28 days after surgery, the rats received intrathecal clonidine (0.3, 1.0, and 3.0 microg) and tizanidine (1.0, 2.0, and 5.0 microg), and the antihyperalgesic effects of thermal and mechanical stimuli were examined. In addition, the changes in blood pressure and heart rate, sedation level, and other side effects after intrathecal administration of drugs were recorded. RESULTS: The administration of 3.0 microg intrathecal clonidine or 5.0 microg tizanidine significantly reversed both thermal and mechanical hyperalgesia. The administration of 3.0 microg intrathecal clonidine, but not 5.0 microg tizanidine, significantly decreased mean blood pressure and heart rate and produced urinary voiding. A greater sedative effect was produced by 3.0 microg intrathecal clonidine than by 5.0 microg tizanidine. CONCLUSION: The antihyperalgesic dose of intrathecal clonidine and the antinociceptive doses produced several side effects. Intrathecal tizanidine at the dose that reversed hyperalgesia would be preferable for neuropathic pain management because of absence of hypotension and bradycardia and lower incidence of sedation. PMID- 12766662 TI - Assessment of differential blockade by amitriptyline and its N-methyl derivative in different species by different routes. AB - BACKGROUND: Increasing the duration of local anesthesia and/or creating greater differential blockade (i.e., selective block of pain-transmitting nerve fibers) has been attempted by modifying currently available agents. Most drugs show a different profile depending on the model or species studied. This study was designed to investigate the differential nerve-blocking properties of amitriptyline and its quaternary ammonium derivative in rats and sheep. METHODS: The Na+ channel-blocking properties of N-methyl amitriptyline were determined with the patch clamp technique in cultured GH(3) cells. Various functions (motor, nociception, proprioception-ataxia) were compared in rats (spinal and sciatic nerve blockade) and sheep (spinal blockade) with amitriptyline, N-methyl amitriptyline, lidocaine, and bupivacaine (partially from historical data). RESULTS: In vitro testing revealed N-methyl amitriptyline to be a potent Na+ channel blocker similar to amitriptyline but with a much longer duration of action. All drug concentrations tested in both the sciatic nerve model and the spinal block model produced no significant differential blockade in rats. Three of six rats in the 20-mM N-methyl amitriptyline group showed residual blockade 4 days after sciatic nerve injection. However, in the sheep spinal model, amitriptyline and in particular N-methyl amitriptyline displayed significant differential blockade at most time points. Sheep data for lidocaine and bupivacaine seemed to be more comparable to the clinical experience in humans than did rat data. CONCLUSIONS: Amitriptyline and N-methyl amitriptyline are potent Na+ channel blockers and show greater differential blockade in sheep than in rats. This differential blockade in sheep is greater than that produced by lidocaine or bupivacaine. PMID- 12766663 TI - Variability in surgical caseload and access to intensive care services. AB - BACKGROUND: Variability in the demand for any service is a significant barrier to efficient distribution of limited resources. In health care, demand is often highly variable and access may be limited when peaks cannot be accommodated in a downsized care delivery system. Intensive care units may frequently present bottlenecks to patient flow, and saturation of these services limits a hospital's responsiveness to new emergencies. METHODS: Over a 1-yr period, information was collected prospectively on all requests for admission to the intensive care unit of a large, urban children's hospital. Data included the nature of each request, as well as each patient's final disposition. The daily variability of requests was then analyzed and related to the unit's ability to accommodate new admissions. RESULTS: Day-to-day demand for intensive care services was extremely variable. This variability was particularly high among patients undergoing scheduled surgical procedures, with variability of scheduled admissions exceeding that of emergencies. Peaks of demand were associated with diversion of patients both within the hospital (to off-service care sites) and to other institutions (ambulance diversions). Although emergency requests for admission outnumbered scheduled requests, diversion from the intensive care unit was better correlated with scheduled caseload (r = 0.542, P < 0.001) than with unscheduled volume (r = 0.255, P < 0.001). During the busiest periods, nearly 70% of all diversions were associated with variability in the scheduled caseload. CONCLUSIONS: Variability in scheduled surgical caseload represents a potentially reducible source of stress on intensive care units in hospitals and throughout the healthcare delivery system generally. When uncontrolled, variability limits access to care and impairs overall responsiveness to emergencies. PMID- 12766664 TI - Effects of postoperative, nonsteroidal, antiinflammatory drugs on bleeding risk after tonsillectomy: meta-analysis of randomized, controlled trials. PMID- 12766665 TI - History of the development and evolution of local anesthesia since the coca leaf. PMID- 12766666 TI - Skin injury with the use of a water warming device. PMID- 12766667 TI - Probable meperidine-induced serotonin syndrome in a patient with a history of fluoxetine use. PMID- 12766668 TI - Treatment of excessive bleeding in Jehovah's Witness patients after cardiac surgery with recombinant factor VIIa (NovoSeven). PMID- 12766669 TI - The neuropsychologic deterioration seen in the placebo group may have been a result of rigorous exclusion criteria. PMID- 12766670 TI - Neuroprotective and antiepileptic activities of ketamine in nerve agent poisoning. PMID- 12766671 TI - Possible dangers of discontinuing statins perioperatively. PMID- 12766672 TI - Acute normovolemic hemodilution: the subgroup of patients likely to benefit remains uncertain. PMID- 12766673 TI - Is attenuation of extracellular dopamine increase in the nucleus accumbens the major mechanism by which dexmedetomidine increases the cocaine seizure threshold in rats? PMID- 12766674 TI - Gabapentin: the first preemptive anti-hyperalgesic for opioid withdrawal hyperalgesia? PMID- 12766675 TI - Systemic effects of epidural medications. PMID- 12766676 TI - Need for additional control in studies of epidural outcome. PMID- 12766677 TI - Can't blame bupivacaine. PMID- 12766678 TI - Postoperative sore throat: due to intubation or reflux disease? PMID- 12766679 TI - Can choice of vasopressor therapy affect rostral spread of spinal anaesthetic? PMID- 12766680 TI - Magnetoencephalographic validation parameters for clinical evaluation of interictal epileptic activity. AB - The authors demonstrate that the confidence volume (the spatial volume that encompasses the 95% probability of source localization) of the single equivalent current dipole is helpful in validating magnetoencephalographic epileptic spike mapping. Such mapping involves distinguishing spikes from other neuronal events. The usual criteria for validating dipole fit reliability involve four parameters correlation coefficient (R > or =0.98), goodness of fit (> or =0.95), root mean square magnetic field value (>400 fT), and dipole moment (Q value > 200 nAm)-but other parameters (direction of dipole moment, location of dipole, and confidence volume) can be considered. In 21 patients with epilepsy, the average correlation coefficient for 608 epileptic spikes was 0.99; average goodness of fit, 0.98; average root mean square, 1,198 fT; and the average Q value, 370 nAm. The mean average confidence volume was 0.30 +/- 0.27 cm3. Correlation coefficient values for quiet brain activity were less than 0.90; goodness of fit values, less than 0.85; and confidence volumes were large (>5 cm3); and for noise runs (no subject) they were even larger (>100 cm3), although correlation coefficient values were more than 0.80 and goodness of fit values were more than 0.85. Confidence volumes for noise data are large-for background brain activity even larger-but confidence volumes for epileptic spikes are small. Confidence volume, in conjunction with other parameters, may be a robust parameter for spike selection. PMID- 12766681 TI - Cortical neuromagnetic fields evoked by voluntary and passive hand movements in healthy adults. AB - Neuromagnetic fields were recorded from the left cerebral hemisphere of six healthy right-handed subjects under three different conditions: (1) externally triggered rapid voluntary extension and flexion of the right hand, (2) passive extension and flexion of the right hand, and (3) stimulation of the skin of the right index finger by means of air pressure. Location analysis using the current density analysis did not reveal any differences between motor evoked field I (MEF I) in active and passive movements, and met the maximum of cerebral activation in the contralateral precentral region. In contrast, the sensory evoked field was located clearly in the contralateral postcentral region. Additionally, a significantly shorter latency of MEF I (with respect to movement onset) was observed in flexion compared with extension in both passive and active movements. These results support the assumption that MEF I is generated by cortical activation resulting from proprioceptive, probably muscle spindle, input. The current density analysis has proved to be an appropriate method for investigating movement-related fields. Furthermore, the described method seems to be appropriate for evaluating the processes of cortical reorganization and the influence of neurorehabilitation within longitudinal studies in patients with lesions in motor centers of the brain. PMID- 12766682 TI - Transient inability to distinguish between faces: electrophysiologic studies. AB - It is not known with certainty at which level of face processing by the cortex the distinction between a familiar and an unfamiliar face is made. Subdural electrodes were implanted under the fusiform gyrus of the right temporal lobe in a patient who developed an unusual inability to distinguish differences between faces as part of the epileptic aura ("all faces looked the same"). A cortical region located posterior to the epileptic focus was identified that exhibited a maximum evoked response to the presentation of facial images (N165), but not to objects, scenes, or character strings. Evoked potentials elicited by a variety of visual images indicated that any perturbation away from novel whole-face stimuli produced submaximal responses from this region of the right temporal lobe. Electrical stimulation of this region resulted in an impairment of face discrimination. It was found that presentation of familiar faces (grandmother, treating physician) produced a different response from that observed for novel faces. These observations demonstrate that within 165 msec of face presentation, and before the conscious precept of face familiarity has formed, this cortical region has already begun to distinguish between a familiar and an unfamiliar face. PMID- 12766683 TI - A case of area-specific stimulus-sensitive postanoxic myoclonus. AB - The authors report a case of area-specific stimulus-sensitive postanoxic myoclonus and discuss possible pathophysiology. A 71-year-old man sustained cardiorespiratory arrest that lasted 10 minutes and remained unresponsive. On the first EEG obtained 8 hours after the arrest there was no cerebral electrical activity before stimulation of the trigeminal-innervated areas. Periorbital stimulation was associated with bursts of spike-wave activity and generalized myoclonic jerks, whereas other types of stimulation did not elicit any response. A second EEG obtained 32 hours later showed a nonreactive alpha coma pattern. The patient died 7 days after the arrest. Area-specific stimulus-sensitive postanoxic myoclonus is very rare. The regularity of generalized bursts of spike-wave activity (cortical response) in response to stimulation of trigeminal-innervated areas suggests that the resting EEG electrocerebral silence may have been a result of cortical suppression with disinhibition of stimulus-sensitive brainstem generated myoclonus. PMID- 12766684 TI - Periodic lateralized epileptiform discharges in multiple sclerosis: a case report. AB - The occurrence of epileptiform abnormalities on the EEG in patients with multiple sclerosis (MS) is rare. The following case correlates the clinical, EEG, MRI, and single photon emission computed tomographic (SPECT) findings in a patient with a long history of MS and acute onset of focal motor seizures and confusion. Two routine EEGs, brain MRI, and brain SPECT were performed. The patient was a 44 year-old woman with a long history of clinically definite MS of the relapsing remitting and secondary progressive form with three events of focal motor seizures followed by generalized tonic-clonic seizures and postictal confusion. The first EEG done during admission showed periodic lateralized epileptiform discharges in the right temporal region. Brain MRI done several weeks later showed scattered T2 hyperintensities in several locations, including the periventricular and subcortical white matter bilaterally. Brain SPECT using Tc99 Neurolite demonstrated decreased perfusion on the right parietal and temporal lobes. This case suggests that focal motor seizures and a transient state of altered consciousness can be the result of an exacerbation of MS. The neurophysiologic expression of these clinical manifestations may present as periodic lateralized epileptiform discharges on the EEG and decreased regional perfusion on brain SPECT. PMID- 12766685 TI - Impact of somatosensory evoked potential monitoring on cervical surgery. AB - Controversy still exists about the necessity of somatosensory evoked potential (SSEP) monitoring during cervical surgery. The purpose of this prospective study is to determine the impact of SSEP monitoring on anterior cervical surgery. Intraoperative SSEP monitoring was performed in 100 patients treated by an anterior cervical approach. The patients were divided into three groups according to their preoperative clinical condition. Somatosensory evoked potential monitoring was performed during five stages of the procedure: M1, after the induction of anesthesia; M2, during positioning; M3, during distraction of the intervertebral space; M4, throughout decompression; and M5, during graft placement. Normal SSEPs were obtained during M1 from all the patients in group 2. Pathologic SSEPs were recorded at M1 in 45 patients from group 1. No SSEPs were recorded at M1 in six patients in group 3. A deterioration of the SSEPs was observed in 35 patients during M2. Deteriorated SSEPs were observed during M3 in 14 patients. No deterioration of the SSEPs was recorded during M4. Intraoperative SSEP monitoring is easy to perform and helps to increase safety during anterior cervical surgery. Critical phases of the surgical procedure were identified and the surgical strategy was modified as a result of this study. PMID- 12766686 TI - Does preoperative paresis influence intraoperative monitoring of the motor cortex? AB - Intraoperative monitoring of motor function by means of motor evoked potentials (MEPs) is a new method. The current study examines the influence of preoperative paresis on the feasibility and reliability of this method. Intraoperative monitoring of MEPs was performed in 58 patients during surgery in the central region. The patients were divided into three groups according to their preoperative strength (group I, muscle strength less than or equal to grade 4 according to the British Medical Research Council grading system [n = 17]; group II, normal strength (n = 36); and group III, muscle strength less than grade 5 but not worse than grade 4 [n = 5]). The motor cortex was stimulated directly with a high-frequency monopolar anodal train. In groups II and III, MEPs were elicited in all patients on cortical stimulation, whereas in group I a response was obtained in only 88% of patients. The MEP parameters in all groups had a broad interindividual range of variation. A correlation between individual intraoperative potential changes and surgical maneuvers was observed in seven patients in group II and in four patients in group I. No MEP changes were recorded in group III. Irreversible MEP changes (groups I and II) resulted in postoperative clinical deterioration. No postoperative deterioration of motor function was observed in patients with reversible MEP changes. Preoperative paresis reduces the feasibility of the method; however, it has no influence on the intraoperative pattern and reaction of the MEPs. PMID- 12766687 TI - Test-retest reliability of chemosensory evoked potentials. AB - This study investigated the test-retest reliability of chemosensory event-related potentials in humans. Olfactory event-related potentials and chemosomatosensory event-related potentials were evaluated in 20 healthy, normosmic subjects. Phenyl ethyl alcohol (PEA, 40% v/v) and H(2)S (4 ppm) served as olfactory stimuli whereas CO(2) (60% v/v) was the chemosomatosensory stimulus. Fifteen stimuli of each compound were applied to each nostril. Identical stimulation sequences were used during three test sessions. Sessions 1 and 2 were separated by a mean of 6.8 days; sessions 2 and 3 by 12.5 days. Electroencephalographic recordings were made from Fz, Cz, Pz, C3, and C4. Amplitudes and latencies of P1, N1, P2, N2, and P3 were measured. Pearson's correlation coefficient was calculated to test the test retest reliability, and the general linear model examined the differences. Most correlations ranged between 0.4 < r < 0.75. Latencies correlated significantly better (P = 0.008) between sessions than amplitudes, even though with CO(2) stimulus amplitudes correlated significantly better than with PEA (P = 0.006) or H(2)S (P = 0.003). No differences arose between measurements from different nostrils for any stimulus. Chemosensory event-related potentials show good test retest reliability. Carbon dioxide amplitudes exhibit better signal-to-noise ratios than PEA or H(2)S amplitudes. Chemosensory event-related potentials are a clinically valuable objective and are reproducible. PMID- 12766688 TI - Single-unit analysis of the spinal dorsal horn in patients with neuropathic pain. AB - Despite the key role played by the dorsal horn of the spinal cord in pain modulation, single-unit recordings have only been performed very rarely in this structure in humans. The authors report the results of a statistical analysis of 64 unit recordings from the human dorsal horn. The recordings were done in three groups of patients: patients with deafferentation pain resulting from brachial plexus avulsion, patients with neuropathic pain resulting from peripheral nerve injury, and patients with pain resulting from disabling spasticity. The patterns of neuronal activities were compared among these three groups. Nineteen neurons were recorded in the dorsal horns of five patients undergoing DREZotomy for a persistent pain syndrome resulting from peripheral nerve injury (i.e., nondeafferented dorsal horns), 31 dorsal horn neurons were recorded in nine patients undergoing DREZotomy for a persistent pain syndrome resulting from brachial plexus avulsion (i.e., deafferented dorsal horns), and 14 neurons were recorded in eight patients undergoing DREZotomy for disabling spasticity. These groups were compared in terms of mean frequency, coefficient of variation of the discharge, other properties of the neuronal discharge studied by the nonparametric test of Wald-Wolfowitz, and the possible presence of bursts. The coefficient of variation tended to be higher in the deafferented dorsal horn group than in the other two groups. Two neurons displaying burst activity could be recorded, both of which belonged to the deafferented dorsal horn group. A significant difference was found in term of neuronal behavior between the peripheral nerve trauma group and the other groups: The brachial plexus avulsion and disabling spasticity groups were very similar, including various types of neuronal behavior, whereas the peripheral nerve lesion group included mostly neurons with "nonrandom" patterns of discharge (i.e., with serial dependency of interspike intervals). PMID- 12766689 TI - Contralateral reinnervation of midline muscles in nonidiopathic facial palsy. AB - The purpose of this study was to analyze contralateral reinnervation of the facial nerve in eight patients with complete facial palsy after surgery or trauma and seven healthy volunteers. All patients had contralateral reinnervation of facial muscles as demonstrated by electrical nerve stimulation versus none of the control subjects. Four patients had facial muscle movements at the site of the damaged nerve. In one patient this was entirely the result of contralateral reinnervation, whereas the other three patients had innervation both ipsilaterally and contralaterally. This implies that renewed facial muscle activity should be examined considering the origin of the reinnervation, either contralateral or ipsilateral. Contralateral reinnervation is a common phenomenon after total facial palsy and can occur alongside ipsilateral reinnervation. It can be mistaken for adequate reinnervation of the damaged nerve, causing postponement of dynamic reconstruction therapy. PMID- 12766691 TI - Cognitive-behavioral therapy for panic disorder: a review of treatment elements, strategies, and outcomes. AB - This article provides an overview of cognitive-behavioral therapy (CBT) for panic disorder. CBT is currently considered a first-line treatment for panic disorder. It offers benefit after short-term intervention, typically consisting of 12-15 sessions conducted in either an individual or a group format. The treatment focuses on the elimination of the patterns that underlie and perpetuate the disorder. Through CBT, patients learn about the nature of the disorder and acquire a set of strategies that counter the fears of panic attacks themselves, and break the recurring cycle of anticipatory anxiety, panic, and agoraphobic avoidance. The collaborative format of treatment, and a focus on elimination of core fears may be factors in enhancing longer-term outcome. In this article, we review the efficacy of CBT as a first-line treatment, a strategy for medication nonresponders, a replacement strategy for patients who wish to discontinue pharmacotherapy, and a potential preventive strategy for at-risk individuals. We also discuss some of the complex issues involved with combination-treatment strategies. PMID- 12766690 TI - How well does cognitive-behavioral therapy treat symptoms of complex PTSD? An examination of child sexual abuse survivors within a clinical trial. AB - Are brief cognitive-behavioral treatments for posttraumatic stress disorder (PTSD) also effective for the wider range of symptoms conceptualized as complex PTSD? Female rape victims, most of whom had extensive histories of trauma, were randomly assigned to cognitive-processing therapy, prolonged exposure, or a delayed-treatment waiting-list condition. After determining that both types of treatment were equally effective for treating complex PTSD symptoms, we divided the sample of 121 participants into two groups depending upon whether they had a history of child sexual abuse. Both groups improved significantly over the course of treatment with regard to PTSD, depression, and the symptoms of complex PTSD as measured by the Trauma Symptom Inventory. Improvements were maintained for at least 9 months. Although there were group main effects on the Self and Trauma factors, there were no differences between the two groups at posttreatment once pretreatment scores were covaried. These findings indicate that cognitive behavioral therapies are effective for patients with complex trauma histories and symptoms patterns. PMID- 12766692 TI - Long-term efficacy of exposure and ritual prevention therapy and serotonergic medications for obsessive-compulsive disorder. AB - What is the long-term outcome of patients with obsessive-compulsive disorder (OCD) who are treated with exposure and response (ritual) prevention (EX/RP) alone, serotonergic medications alone, or their combination? How is the long-term outcome of these patients affected by the discontinuation? Follow-up assessments were conducted with 62 patients treated for OCD an average of 17 months posttreatment (range: 6-43 months). Patients received one of three treatments: serotonergic medications (fluvoxamine or clomipramine), intensive behavior therapy involving EX/RP, or intensive EX/RP with concurrent antidepressant medication. At follow-up, no differences in OCD symptom severity were found among the three treatment groups. However, when current medication use was taken into consideration, differences among the three treatment groups emerged. Among patients who were medication-free at the time of follow-up assessment (n=37), those in the EX/RP-alone and EX/RP-with-medication groups had lower symptom severity ratings than those in the medication-only group on 4 out of 6 measures. There were no differences in OCD severity ratings among patients taking medications at follow-up (n=25). Although these findings are interpreted with caution due to the uncontrolled nature of the study, results suggested that long term outcome may be superior following EX/RP than following serotonergic medications, after discontinuation. For patients who remain on medications, the treatment produced benefits equivalent to EX/RP. PMID- 12766693 TI - Cognitive-behavioral therapy for social anxiety disorder: supporting evidence and future directions. AB - The present paper examines the role of cognitive-behavioral therapy (CBT) in the treatment of social anxiety disorder (SAD). A cognitive-behavioral model of SAD is first presented. Different modalities of CBT for SAD are then described, including exposure, cognitive restructuring, relaxation training, and social skills training, and evidence supporting their efficacy is reviewed. The comparative and combined impact of CBT and pharmacotherapeutic interventions is also explored. CBT appears to be an efficacious treatment for SAD. However, the overall efficacy CBT may be increased by closer examination of the active ingredients of treatment. Such analyses may also enable more successful integration of the different CBT techniques and of CBT and pharmacotherapy in the treatment of SAD. PMID- 12766694 TI - Cognitive-behavioral therapy for generalized anxiety disorder with integrations from interpersonal and experiential therapies. AB - After providing background information on the definition and nature of generalized anxiety disorder, this article describes cognitive-behavioral therapy (CBT) methods that have been empirically supported in the treatment of this disorder. Subsequent to this description, relevant outcome literature is briefly reviewed, along with evidence that the addition of other techniques beyond traditional CBT methods may be necessary to maximize clinical outcome. A description is then provided of an integrated interpersonal/emotional processing therapy that the authors have recently added to their CBT protocol. CBT with and without this integrated treatment is currently being evaluated in an experimental trial. PMID- 12766695 TI - Neurocognitive Manifestations of Human Immunodeficiency Virus. AB - Is human immunodeficiency virus still a terminal condition? Recent advances in treatment have significantly reduced both mortality and morbidity associated with HIV, but these treatments have not been successful in eradicating the virus itself. As such, HIV has evolved into a chronic condition that is complicated by neurocognitive factors. Cognitive difficulties associated with HIV are characterized by a subcortical pattern with primary deficits in information processing speed and psychomotor speed. These deficits interfere with the ability of patients to complete important instrumental activities of daily living even in the absence of dementia. Treatment of HIV improves neurocognitive functioning, but the regimens are complex and patient adherence is critical. Cognitive factors can negatively impact treatment adherence, which in turn results in poorer immunological, cognitive, and psychiatric outcome. This cycle emphasizes the important interrelationships between symptom expression and treatment outcome in patients with HIV. The nature of these relationships will change with further developments in treatment regimens such as once-daily dosing. Less complex treatment approaches should improve health outcome as well as provide additional opportunities to further understand the impact of HIV on brain function. PMID- 12766696 TI - Substance P and Human Immunodeficiency Virus Infection: Psychoneuroimmunology. AB - Effects on the immune system caused by changes in behavioral state or brain activity are mediated, at least in part, through neuroendocrine-immune pathways. Life stress and depression may be associated with altered blood levels of central nervous system-released neuropeptides, including substance P (SP). SP acts as a neuroregulator or neurotransmitter in the conduction of nociceptive stimuli, and is a modulator of neuroimmunoregulation. This review summarizes current knowledge regarding the role of the neuropeptide, SP, in psychoneuroimmunology, in particular as it relates to human immunodeficiency virus infection and acquired immunodeficiency disease syndrome. The association between depression, anxiety, and stress in HIV-disease progression suggests that neurobiologic and neurophysiologic factors play a role in modulating HIV infection and responses to antiretroviral therapy. Individuals with HIV or AIDS may experience stressful life circumstances that can result in increased symptoms of anxiety, stress, and/or depression. Furthermore, psychological and psychiatric symptoms, which occur in individuals with HIV and AIDS, may be related to the progression of AIDS disease. This review presents evidence from the literature, as well as findings from basic investigations conducted in the authors' laboratories, demonstrating that SP may play an important role in HIV pathophysiology. SP can impact the susceptibility of immune cells to HIV infection and modulate immune cell functions in ways that may affect the course of HIV in infected individuals. Moreover, modulation of SP activity and SP receptor is being explored for its potential as a novel therapeutic approach to the treatment of some psychological and psychiatric disorders and to the design of new anti-HIV therapy. PMID- 12766697 TI - Literary Factitious Epilepsy Syndromes. AB - Several factitious epileptic syndromes have been associated with famous literary characters. While these syndromes include symptoms other than pseudoseizures, and while pseudoseizures can occur in other syndromes, a review of these disorders provides insights into factitious seizures and epilepsy. PMID- 12766698 TI - An Unusual Case of Motor and Vocal Tics With Obsessive-Compulsive Symptoms in a Young Adult With Behcet's Syndrome. AB - Behcet's disease is a chronic relapsing-remitting, multi-system inflammatory disorder of unknown etiology, characterized by recurrent oral and genital ulcers, uveitis, and skin lesions. The most common central nervous system findings are demyelization and perivascular cell infiltration; meningoencephalitis, cerebral atrophy, and vascular thrombosis may also occur. Neuropsychiatric symptoms may include: paroxysmal dystonia, parkinsonism, delirium, hallucinations, and behavioral disturbances. We report an unusual case of explosive onset of motor and vocal tics with obsessive-compulsive symptoms in a 22-year-old male with Behcet's disease. The putative roles of infection and autoimmune mechanisms in the pathogenesis of tics and obsessive-compulsive symptoms are explored. PMID- 12766699 TI - Global Benefit-risk Evaluation of Antidepressant Action: Comparison of Pooled Data for Venlafaxine, SSRIs, and Placebo. AB - Do antidepressants have an equivalent risk-benefit ratio? Venlafaxine, a serotonin-norepinephrine reuptake inhibitor, is an effective antidepressant for treating major depression. The results of some clinical studies have suggested that venlafaxine may have more potent efficacy in sustaining remission in patients with major depression. Comparative clinical studies, however, lack suitable power to discern treatment differences in safety and also lack a quantitative basis for comparing risk and benefit. A global benefit-risk analysis of pooled data from eight randomized, double-blind, clinical trials of the safety and efficacy of venlafaxine and selective serotonin reuptake inhibitors (SSRIs) was performed. By using the ratio measure of risk-benefit, patients treated with venlafaxine (n=851) for 6-8 weeks experienced a relative gain of 1.57 compared with SSRI-treated patients (n=743) and a relative gain of 2.27 compared with placebo-treated patients (n=439). Subgroup analyses showed a relative gain of 1.35 for venlafaxine-treated patients (n=538) compared with fluoxetine-treated patients (n=549) and a relative gain of 2.53 compared with placebo-treated patients (n=357). A dose-response relationship was apparent between low (<75 mg/day), medium (75-150 mg/day), and high (>150 mg/day) dosages of venlafaxine; r values were 0.758, 0.822, and 1.181, respectively (P=.023, high dosage versus placebo; P=.030, medium dosage versus placebo). Important differences in risk and benefit exist between venlafaxine and SSRIs as a group compared with fluoxetine alone. A significant gain in benefit-risk in the treatment of major depression was observed with an increase in venlafaxine dosage from 75->150 mg/day. PMID- 12766700 TI - Insect repellents. PMID- 12766701 TI - Ophthalmic cyclosporine (Restasis) for dry eye disease. PMID- 12766702 TI - Xanax XR for panic disorder. PMID- 12766703 TI - [SIAARTI recommendations for the admission and discharge from Intensive Care and for the limitation of treatments in Intensive Care]. PMID- 12766704 TI - Clinical decision making during resuscitation and guidelines for termination of the resuscitation effort. AB - Cardiopulmonary resuscitation (CPR) is the appropriate intervention to sustain respiratory and circulatory functions in patients with diseases that are potentially reversible; while CPR is unequivocally inappropriate for patients with terminal diseases when there is a consensus of medical opinion that is no reasonable likelihood of meaningful survival. In practical application patients and their families seek guidance based on understanding. CPR when employed without expectations of benefit, will be likely to further compromise neurological function, produce iatrogenic injury, and add discomfort to the dying patient. The decision not to resuscitate remains a reasoned medical judgement. In some condition, transferring the patient with terminal diseases to an alternate physician or institution is ultimately respectful both for the patient and of physician. PMID- 12766705 TI - When the ICU patient refuses vital supports. AB - The aim of this paper is to discuss the problem of caring for ICU patients who request forgoing of intensive supports; in particular, evaluating their competence and building effective relationships among the patients themselves, their relatives and the health care team. The histories of 2 adult competent ICU patients 2 asked that vital intervention be forgone are presented, together with the discussion of problems posed by such a request and of possible solutions which respect patients' autonomy and bystanders needs. These patients were hospitalized in an Italian general 14 bed ICU. The patients' request were weighted against their prognosis and their view of life, in order to evaluate their reliability. Also the refusal of either treatment or outcome was evaluated, in order to effectively safeguard the patients' interests. The request of one patient was satisfied and he was allowed to die, after all the involved people had understood and accepted the final decision. The second patient, who was somehow forced to undergo intensive treatment, is alive and satisfied with having been cured. In Italy too, autonomy is an increasingly applied principle in end-of life decisions in ICUs. It is usually tempered by a consideration about the patients' best interest as perceived by involved bystanders. PMID- 12766706 TI - [Actions to undertake after an accident]. PMID- 12766707 TI - How to recruit the injured lung. AB - Alveolar recruitment represents a challenging issue in ALI/ARDS patients. Multiple techniques have been compared: intermittent sighs, sustained application of high pressure in single or multiple episodes, use of progressive higher PEEP and lower tidal volumes (VT), with a fixed upper limit, and increase of PEEP, without modifying VT. Encouraging results emerge also from the use of prone position, that allows a better distribution of transalveolar forces, thus reducing ventilator induced lung injury. Moreover the use of spontaneous breathing, such as Bi-PAP mode, enhances re-expansion of dorsal lung regions and intriguing, but still uncertain results derive from biological variability of ventilatory pattern. Finally, a pressure-volume (P-V) curve of respiratory system can be employed to set appropriate PEEP level, to prevent collapse of new recruited alveoli. To monitor alveolar recruitment we can use P-V curves, continuous intra-arterial gas analysis, electrical impedence tomography. It is worth noting that different recruiting techniques are characterised by different efficacy and adverse hemodynamic effects. In conclusion, The "Open lung" approach should not be applied to every patient; it should be reserved to restore lung volume if deterioration occurs, by means of adequate PEEP level and lowest acceptable FiO(2). PMID- 12766708 TI - Arterial blood pressure monitoring. PMID- 12766709 TI - [Fever in intensive care: an open problem]. AB - Fever is one of the most aspecific marker of disease, it is considered a protective host defense response and it is the result of a reset of the hypothalamic thermostat. Fever is a common problem in ICU patients and it is associated with infective and non infective causes. Fever presenting in ICU should always be a source of concern and the first and immediate priority is to determine its clinical significance. PMID- 12766710 TI - Treatment of fever in neurosurgical patients. AB - Even moderate temperature elevations soon acute cerebral damage may markedly worsen initial brain injury. These effects may justify aggressive antipyretic treatment in neurosurgical intensive care unit (NICU). On the basis of a literature survey, it is observed that fever is extraordinarily common in the neurosurgical intensive care unit during the acute phase of subarachnoid hemorrhage, stroke, and traumatic brain injury. Several clinical studies also suggest worsened neurologic outcome in patients who are febrile compared to those who are not. Pyrexia is more frequent in infected than noninfected patients. Infections (mainly in the respiratory tract) are usually diagnosed in the majority of febrile NICU patients. Laboratory investigations are quite clear regarding the adverse effects of fever in terms not only of functional outcomes, but also histological and neurochemical injury. Even though fever may cause diagnostic confusion (central fever vs infectious), the potentially devastating effects of pyrexia in patients with cerebral diseases may proceed to treat in any case. An attempt to correct fever appears warranted in all patients with acute cerebral damage in order to obtain a better functional recovery and to limit maximally any further insult to the brain. Some of the more common and innovative methods to control body temperature in order to mitigate the detrimental effects of pyrexia following acute neurological injury are explored. Maintenance of normothermia appears to be a desirable therapeutic goal in managing the patients with damaged or at-risk brain tissue. However, it has not been established conclusively that the benefits of antipyretic therapy outweigh its risks and that despite a sound physiologic argument for controlling fever in the brain-injured patient, there is no evidence that doing so will improve their outcome. PMID- 12766711 TI - [Neuro-link, an Italian traumatic coma data bank: what did we learn from the first 1000 patients and how can we do better? ]. AB - To understand the complex physiopathology of post-traumatic brain damage is important to have data on epidemiology, clinical course, monitoring, effect of therapy and outcomes. In 1997 3 neuro-intensive care units in the Milan metropolitan area developed a computer assisted database named Neuro-Link (NL) for collection of information on head injury. All head injured patients requiring intensive care during the first 24 hours post-trauma were eligible. The data collection form was designed for use with a computer interface to cover: 1) general, previous and admission data; 2) secondary insults and complication; 3) CT scan and monitoring data; 4) outcome data. Two different data collections were performed: 1) NL domestic (data from 3 centres from 1997); 2) NL 18 centres (3 month survey of Italian centres with interest in neurotrauma care). An audit of the data was performed. NL domestic included 1 085 patients from 1997 to 2002. NL 18 centres included 282 patients in the 3 month period. Audit is performed on 35 000 data per year.A large number of good quality information on head injury patients is now available. The database is useful for: 1) production of information; 2) base for prospective studies. PMID- 12766713 TI - [How to quantify the severity of brain injury during intensive care after adult head trauma]. AB - Adequate early assessment of brain damage is essential. Location, extension and severity of structural damage affect brain function and ultimately determine the outcome. The extent of functional impairment, and the morphology of intracranial lesions, require specific treatment, often a combination of medical and surgical interventions. Brain damage usually evolves over time, and repeated assessments are necessary. Clinical evaluation is often biased by concomitant sedation and/or anesthesia, but remains necessary. A revision of the literature is presented. Brain damage is assessed combining clinical and instrumental data. Clinical examination is performed assessing the 3 components of the Glasgow Coma Scale. Spontaneous or stimulated (pain stimulus) eye opening, verbal and motor responses are observed after hemodynamic and respiratory stabilisation. Unfortunately a significant proportion of patients can not be properly examined for several reasons: eye opening can be altered by palpebral and facial injuries, verbal response can be impaired by maxillo-facial injuries or by endotracheal intubation, and motor response remains the most consistent parameter. Sedation, analgesia and myorelaxants, however, can profoundly diminish or abolish the motor response to maximal stimulation, so that examination should be performed after clearance of drugs. Often alcohol or other substances can further impair the neurological performances. Pupils diameter and reactivity to light should be observed, excluding pharmacologic effects (as dilation due to catecholamines) and direct ocular or orbital damage. The CT scan is necessary for disclosing surgical masses and for identifying the extent of diffuse damage and the location of focal lesions. These data should be combined with additional functional exploration, as provided by cerebral extraction of oxygen and electrophysiologic data. Early estimation of cerebral damage is complex and prone to mistakes. Accurate, repeated evaluations, based on the combination of clinical observation and imaging, are necessary. PMID- 12766712 TI - [Brain vulnerability and its modulation]. AB - Clinical and experimental studies revealed that the injured brain is highly vulnerable to a subsequent insult. Surfery of the literature pertinent to clinical and experimental traumatic brain injury (TBI) is made. Increased vulnerability of the traumatically injured brain to an additional sub lethal ischemic, hypoxic, excitotoxic, or mechanical insult has been clearly demonstrated. Compared to traumatic brain injury alone, the double insult paradigm dramatically increases the brain damage. Brain vulnerability following TBI can be explained by a reduced ability to compensate for a reduction of cerebral blood flow (CBF) and oxygen (O(2)) delivery to the brain or inability to meet an increased metabolic demand. In addition, there is a specific increased sensitivity to delayed insults induced by the first injury. Potential mechanisms of the increased sensitivity to a second insult might be related to post traumatic gene expression alterations leading to changes in neurotransmitters release, density of receptors and reduced thresholds for activation of pathways leading to delayed cell death. The brain is vulnerable to repetitive injuries. Derangements of compensatory mechanisms are responsible, in part, for this vulnerability. Additional work is needed to better understand the molecular pathways leading to secondary damage and to find novel therapeutic strategies to modulate the brain response to TBI. PMID- 12766714 TI - Targets for resuscitation from shock. AB - Resuscitation from circulatory and respiratory failure represent mainstays of emergency and critical care management. Importantly, no amount of resuscitative effort will be successful in promoting patient survival if the primary reason for the shock state is not identified and treated, independent of resuscitation. Having said that, aggressive resuscitation to normal functional levels of blood flow and organ perfusion pressure during the first 6 hours following the development of shock improves outcome both in patients with trauma or sepsis. However, clinical studies have demonstrated that restoration of total blood flow to supranormal levels in subjects with established shock that has been present for over 6 hours does not improve survival. Still, some defined clinical targets are essential in these patients as well to prevent further organ injury due to ischemia and its associated inflammatory response. Thus, the rapid restoration of normal hemodynamics by conventional means, including fluid resuscitation and surgical repair, results in a better log term outcome than inadequate or delayed resuscitative efforts. Clear initial targets for resuscitation are a mean arterial pressure > 60 mm Hg, and a cardiac output and O(2) transport to the body adequate enough to prevent tissue hypoperfusion. The level of cardiac output needed to achieve this goal is probably different among subjects and within subjects over time. Indirect signposts of adequate perfusion, such as venous O2 saturation, mentation, urine output and local measures of tissue blood flow are useful in monitoring this response. PMID- 12766715 TI - Genetic variation and risk of sepsis. AB - Sepsis is the leading cause of death in non-coronary intensive care unit patients. Sepsis is caused by the immune response to infection and is manifest by pain, fever and edema as the result of the activation of coagulation and inflammatory responses. In severe cases, sepsis leads to organ dysfunction and failure. Sepsis affects more than 750,000 people each year in the US alone, with a mortality rate of over 35 percent making it one of the leading causes of death in developed countries. In addition many patients that die of other diseases have their hospital courses complicated by sepsis. Most patients with infection do not develop severe sepsis and septic shock and yet those that do have a significantly increased risk of death. Genetic and environmental variables may influence why one patient with infection gets sicker than the next. For example, people may be programmed to respond to infection in different ways; some with aggressive immune responses that may be able to wipe out infection before it manifests itself in physical symptoms, while others may have less aggressive immune systems that allow them to get sick more often. The discovery of various common genetic polymorphisms in genes that control the inflammatory response (e.g. tumor necrosis factor) has lent credence to this hypothesis. Yet discovery of the actual relationship between risks of infection / severe sepsis and individual genotypes will require larger, more rigorously designed studies. PMID- 12766716 TI - Sepsis clinical knowledge: a role of steroid treatment. AB - After a period of initial enthusiasm, several trials cast serious doubts on the usefulness of corticosteroids for the treatment of patients with severe sepsis. Short course with high doses of steroides should not be given in patients with severe sepsis. The attention is now addressed to low-dose of corticosteroides. The rational for a replacement therapy with hydrocortisone in patients with cathecolamines-dependent septic shock is based on the concept that this may be complicated by an occult adrenal insufficiency and a glucocorticoid peripheral resistance syndrome. Low doses of hydrocortisone has been shown to reproduce the normal effects of cortisol: anti-inflammatory properties and an increased in the vasoconstrictor response to cathecolamines. There is no concordance in literature about the role of replacement therapy with hydrocortisone on survival in patients with septic shock. Waiting for the results of the European confirmatory phase III trial, and based on the results of the French phase III trial, one may recommended to treat septic shock patients who have a cortisol increment after ACTH of less than 9 micro g/dl with 50 mg of hydrocortisone every 6 hours for seven days combined with 50 micro g of fludrocortisone once a day for seven days. PMID- 12766717 TI - Levosimendan compared with dobutamine in low output patients. AB - There are 2 studies which have investigated the hemodynamic efficacy of levosimendan compared to dobutamine in congestive heart failure patients. The first is a dose finding comparative 24-h infusion trial which included 95 NYHA II III patients to different doses of levosimendan and 20 patients to dobutamine administered as a continuous, open-label infusion of 6 microg/kg/min. Efficacy and safety of levosimendan in severe low-output heart failure a randomized, double-blind comparison to dobutamine study compared the short- and long-term efficacy and safety of a single 24-hour infusion of levosimendan (n=103) with dobutamine (n=100) in hospitalised patients in acute heart failure. This double blind, parallel-group, randomised trial showed that, irrespective of the aetiology of the heart failure, levosimendan produced significantly greater improvement in major determinants of cardiac function in heart failure patients compared to dobutamine. Levosimendan produced significantly greater haemodynamic improvements than dobutamine by significantly reduced mortality at 31 days compared with dobutamine; this reduction was maintained at 180 days. Levosimendan significantly increased the number of days alive and out of hospital, compared with dobutamine. It was better tolerated than dobutamine and fewer patients receiving levosimendan experienced arrhythmias and myocardial ischaemia, compared with dobutamine. Levosimendan produced haemodynamic responses that were unaffected by concomitant use of beta blockers. PMID- 12766718 TI - Nutritional and pharmacological support of the metabolic response to injury. AB - Severe burn incites metabolic disturbances which last up to one year post injury. Persistent profound catabolism after severe burn hampers rehabilitative efforts delaying meaningful return of individuals to society. The simplest effective anabolic strategies for severe burn injuries are early excision and grafting of the burn wound, prompt treament of sepsis, maintenance of environmental temperature at 30-32 inverted exclamation mark C, continuous enteral feeding of a high carbohydrate, high protein diet, early institution of vigorous resistive and aerobic resistive exercise programs. To further minimize erosion of lean body mass administration of recombinant human growth hormone, insulin, oxandrolone or propranolol are all reasonable approaches. Exogenous continuous low dose insulin infusion, beta blockade with propranolol and the use of the synthetic testosterone analog, oxandrolone are the most cost effective and least toxic pharmaco therapies to date. PMID- 12766719 TI - [Nutritional therapy in surgical patients: an update]. AB - Patients undergoing major gastrointestinal surgery, often require an adequate artificial nutritional (AN) support for a pre-existing state of malnutrition and/or to overcome forced periods of postoperative starvation and/or for complications that alter the host metabolic response. When an indication to AN is given, enteral feeding should be preferred to parenteral nutrition because more physiological and less expensive. Moreover, recent data showed that patients fed enterally, rather than parenterally, in the postoperative period, have a significant better outcome with a reduction of morbidity and hospitalisation. The supplementation of standard feeds with key nutrients having immunomodulatory properties, such as arginine, omega-3 fatty acids and glutamine (pharmaconutrients), allows to control effectively the surgery-induced immunosuppression and hyperinflammation. An analysis on the principles of evidence-based medicine, supports the hypothesis that the pre-perioperative use of formulas enriched with pharmaconutrients, significantly reduces the rate of infectious complications and saves health care resources. PMID- 12766720 TI - Lactic acidosis. AB - The detection of tissue hypoxia and its correction is one of the aim of the hemodynamic monitoring. Classical hemodynamic variable often fail to achieve this goal. Lactate measurements may be a good indicator of tissue hypoxia. Selected review of the articles on lactate in critically ill patients. Tissue hypoxia is associated with an increase in blood lactate levels. However lactate can also be produced in aerobic conditions, in inflammated tissues, and lactate clearance is often decreased in critically ill patients. Whatever its origin, blood lactate levels have a strong predictive value. The interpretation of blood lactate levels is difficult. Nevertheless, monitoring blood lactate levels can be useful to detect tissue hypoxia and to monitor the effects of therapy. PMID- 12766721 TI - Stroke volume variations. AB - During the cyclic changes in intrathoracic pressure, changes in stroke volume characterize the state at which both ventricles are preload dependant. Determining stoke volume variations may thus help to predict fluid responsiveness in mechanically ventilated patients. Selected review of the articles having investigated the stroke volume variations in critically ill patients. Stroke volume variations can effectively predict preload dependency in mechanically ventilated patients. Stroke volume can either be measured directly or estimated using the analysis of the arterial pressure waveform. Alternatively, pulse pressure variations can be calculated. However, these index are sensitive to tidal volume. During mechanical ventilation with low tidal volumes, the ventilatory-induced changes in preload may be too small to generate changes in stroke volume, even in preload dependant patients. Stroke volume variations can be useful to detect fluid responsiveness in mechanically ventilated patients. PMID- 12766722 TI - [Vital signs of hemodynamic monitoring]. AB - The aim of hemodynamic monitoring in intensive care is to recognize derangements in physiologic variables, which herald the progression toward organ failure. Traditionally the term "vital signs" refers to heart rate, arterial pressure, respiratory rate and body temperature monitoring. Continuous monitoring of vital signs, is advocated, since trends are more significant than single measurements, and is still a cornerstone, in the hemodynamic evaluation of a critically ill patient. Nevertheless, the spectrum of hemodynamic derangements that can arise during intensive care unit stay is very large and often additional information, beside the vital signs, are needed to evaluate correctly the individual patient. PMID- 12766724 TI - [Infection prevention and control in intravascular devices]. AB - Intravascular devices (IVD) are indispensable in the care of the critical patient; even so, their use can be complicated by infection, which is generally associated with longer hospital stay and ensuing higher hospital costs. It is therefore imperative that guidelines are applied that constitute a basis of information upon which the individual facility can develop its own strategy. The strategy can be outlined under the following points: a) staff training, b) surveillance of IVD-associated infections, c) hand washing, d) barrier measures during catheter introduction and management, e) insertion site management and medication systems for the insertion site, f) choice and replacement of the IVD, g) replacement of intravenous administration devices and liquids, h) antimicrobial prophylaxis. In the management of central venous catheters (CVC), recommendations call for: 1) the use of a single lumen CVC, unless multiple accesses are needed; 2) the peripheral placement of CVCs, both in the use of tunneled catheters and/or implantable vascular devices in patients over 4 years of age in which long-term vascular access (> 30 days) is planned; 3) the use of completely implantable devices in pediatric patients less than 4 years of age requiring long-term vascular access; 4) the use of the subclavian artery as the site of CVC insertion unless clinically contraindicated (e.g. coagulopathy, anatomic alterations); 5) the application of barrier precautions during CVC introduction and in the management of the catheter and the insertion site. PMID- 12766723 TI - [A rationale basis for airways conditioning: too wet or not too wet? ]. AB - Medical gases conditioning during mechanical invasive ventilation is nowadays a problem. In fact, in spite of conditioning guidelines, absolute humidity (AH) into 25-35 mg/l, clinical evaluation of the optimal level of airway humidification has not yet been established with certainty. Physiologically, during spontaneous respiration the airway hydric balance, inspiratory AH expiratory AH, is negative of 27 mg/l about. Usually the patients on mechanical ventilation have an expiratory AH of 32-33 mg/l. An overhumidification of inspired gases, positive airway hydric balance, gives anatomic-physiological alterations of airways and lung parenchyma. During invasive mechanical ventilation, the practice of active hot humidifiers has a positive or level airway hydric balance. We think that inspired AH must be equal to expired AH to maintain an airway hydric balance at least level. At last, the temperature of inspired gases, with active hot humidifiers, shouldn't exceed 32-34 degrees C. PMID- 12766725 TI - [Infection control and prevention in intensive care: environmental aspects]. AB - Surveillance is understood as the monitoring over time of the onset of preventable infection and/or microbial colonization in patients in intensive care units (ICU), where the prevalence of nosocomial infections, particularly those caused by microorganisms multiresistant to antibiotics, is alarmingly high. Surveillance deals with: 1) pathogens responsible for infection and their patterns of antibiotic resistance; 2) epidemics; 3) endemic infections selected according to predictability; 4) frequency of use of specific invasive procedures, where the greater the use, the higher the frequency of nosocomial infections associated with them; 5) use of antibiotics. Surveillance is operated through: 1) the microbiology laboratory, which is responsible for accurate surveillance of hospital infections; 2) the active surveillance on the wards, which concerns: a) identification of infections either on admission or during hospitalization or both, b) use of invasive procedures, control of parameters, and data collection, c) type of patients treated, identification of infected patients and data analysis and interpretation. Infection prevention complementary to surveillance starts with the adoption of strategies and interventions based on the application of universal precautions and/or on the route of transmission. A crucial point is the sterilization and disinfection of medical devices. In recent years there has been a steady rise in the number of infections caused by medical devices, most likely because of the increasing use of sophisticated and complex equipment that is often difficult to decontaminate and because of the rise in the number of immunocompromised patients or those susceptible to infections. PMID- 12766726 TI - [Ventilator associated pneumonia]. AB - Ventilator associated pneumonia (VAP) is a nosocomial lower respiratory tract infection that ensues in critically ill patients undergoing mechanical ventilation. The reported incidence of VAP varies between 9% and 68% with a mortality ranging between 33% and 71%. Two key factors are implicated in the pathogenesis of VAP: bacterial colonization of the upper digestive-respiratory tract and aspiration of oral secretions into the trachea. Preventive measurements are advocated to reduce the incidence of VAP, such as selective decontamination of the digestive tract (SDD), supraglottic aspiration and positioning. Prompt recognition and treatment of established VAP has also been demostrated to affect outcome. Therefore, the knowledge of risk factors associated with the development of VAP and the implementation of strategies to prevent, diagnose and treat VAP are mainstems in the nursing of mechanically ventilated patients. PMID- 12766727 TI - [Role of nurses in the evaluation of septic patients]. AB - ICU nurses hold an important role in the management of septic patients underlining with their ability to recognize SIRS that is the first step in the proinflammatory and procoagulant cascade following an infection. Early and timely approach to organ dysfunction can indeed modify the damages due to hypoperfusion. The ability to recognize organ dysfunction using different monitoring devices available should be part of the nursing attitudes. PMID- 12766728 TI - Long-term survival after resuscitation from cardiac arrest: cause for optimism and continued efforts. PMID- 12766729 TI - Diagnosing and managing acute pulmonary embolism: role of cardiac troponins. PMID- 12766730 TI - What did we learn from the OPTIMAAL trial? What can we expect from VALIANT? PMID- 12766731 TI - Radionuclide viability testing: should it affect treatment strategy in patients with cardiomyopathy and significant coronary artery disease? AB - BACKGROUND: Ischemic heart failure is a significant source of morbidity and mortality, yet it has an unclear treatment strategy. The assessment of viable myocardium by nuclear imaging studies has shown promise in predicting improvements in ejection fraction and symptoms. However, the relationship of viability to long-term mortality has not been fully established. METHODS: A number of studies have addressed long-term mortality with nuclear viability imaging in patients with impaired left ventricular function and significant coronary artery disease. These studies were analyzed to determine differences in design, results, trends, and limitations. They were then evaluated by use of qualitative criteria established for prognostic studies. RESULTS: Fourteen studies met our criteria. Although the conclusions differed, it appears that patients with viability who undergo revascularization have the highest survival rate, whereas patients with viability who are treated medically have a much lower survival rate. Patients without viability have an intermediate survival rate, regardless of treatment. Several limitations were identified, including a lack of randomization, small sample size, inadequate follow-up, and extensive study protocol and design differences. CONCLUSIONS: The use of viability testing in patients with heart failure and significant coronary artery disease has shown promise in predicting the long-term mortality rate with treatment allocation. However, there is a need for further study involving larger cohorts with a randomized design, longer periods of follow-up, improved study designs, and identification of referral bias and viability prevalence. PMID- 12766732 TI - Brugada syndrome and "Brugada sign": clinical spectrum with a guide for the clinician. AB - BACKGROUND: Patients with the manifest Brugada syndrome have an inordinate risk of sudden death and are candidates for implantation of a defibrillator. The Brugada type electrocardiogram (ECG) abnormality (the "Brugada sign"), however, is known to be associated with a wide range of conditions, many of which may not pose such a threat. Clinicians need guidance in choosing a rational approach for the evaluation and treatment of patients with a finding of the Brugada sign. METHODS: A systematic literature search was performed to identify publications on the Brugada syndrome and the Brugada-type ECG abnormality, with special emphasis on analyzing outcomes data. In addition, the ECG database of our institution was reviewed for tracings consistent with the Brugada sign, and, when possible, clinical correlations were made. RESULTS: Patients with the Brugada sign and a family history of sudden death or a personal history of syncope are at a high risk of sudden death and therefore should be strongly considered for implantation of a defibrillator. In patients who are hospitalized and critically ill, the Brugada sign is frequently the result of severe hyperkalemia, drug toxicity, or right ventricular injury. In most individuals with no symptoms and without a family history of sudden death, the Brugada sign is likely a normal variant. CONCLUSIONS: Most patients with the Brugada sign can be risk-stratified with simple clinical tools. Specific testing for the Brugada syndrome should be reserved for questionable cases and for the research setting. A provisional diagnostic-therapeutic algorithm is offered as a means of assisting the clinician in the evaluation and treatment of patients with the Brugada sign. PMID- 12766733 TI - Circadian rhythm in the cardiovascular system: chronocardiology. AB - BACKGROUND: We reviewed recent progress in the study of the chronobiological aspects of the cardiovascular system. METHODS: Medline was used as the main search tool, and the full texts of selected papers were obtained. RESULTS: More than 300 references were found, and 52 of them, representing the major findings in this field, were included in the reference list. Results of these studies confirm that most cardiovascular physiological parameters (such as heart rate, blood pressure, electrocardiogram indices) and pathophysiological events (myocardial ischemia/infarction, sudden cardiac death) show circadian rhythms. Results also suggest that consideration of these rhythms is important for the diagnosis and treatment of cardiovascular disorders and that restoration of normal circadian rhythms may be associated with clinical improvement. CONCLUSION: The study of circadian rhythms in the cardiovascular system is emerging as an important area of investigation because of its potential implications for patient management. PMID- 12766734 TI - Atrial stunning: determinants and cellular mechanisms. AB - BACKGROUND: Atrial stunning is a transient depression of atrial and atrial appendage mechanical function after successful cardioversion of atrial fibrillation compared with its precardioversion state. METHOD: Atrial stunning associated with different methods of cardioversion of atrial fibrillation and the determinants and cellular mechanisms of atrial stunning were elaborated by thoroughly examining the studies on the subject identified through a comprehensive literature search. RESULTS AND CONCLUSION: Atrial stunning has been reported with all methods of cardioversion of atrial fibrillation, including transthoracic electrical, low-energy internal electrical, pharmacological, and spontaneous. It is a function of the underlying atrial fibrillation becoming apparent at the restoration of sinus rhythm, regardless of the method used for conversion. Unsuccessful cardioversion does not result in atrial stunning. The duration of the preceding atrial fibrillation, atrial size, and underlying structural heart disease are the determinants of atrial stunning. A shorter duration of atrial fibrillation and smaller atrial diameters are associated with a relatively less severe stunning, lasting for a shorter duration. Atrial stunning after cardioversion of atrial fibrillation of <1 week usually resolves within 24 hours, and atrial stunning after cardioversion of chronic atrial fibrillation usually resolves within 4 weeks. Tachycardia-induced atrial cardiomyopathy, atrial cytosolic calcium alterations with down-regulation of the L-type Ca2+ channels and up-regulation of the Na+/Ca2+ exchanger, atrial hibernation with myocyte dedifferentiation and myolysis, and atrial fibrosis are the suggested mechanisms underlying atrial stunning. Atrial stunning determines the risk of postcardioversion thrombus formation in atria and atrial appendages, the duration of postcardioversion anticoagulation therapy, the recovery of the atrial contribution to the ventricular function, and the functional recovery of the patients after successful cardioversion of atrial fibrillation. PMID- 12766735 TI - Automated three-dimensional assessment of coronary artery anatomy with intravascular ultrasound scanning. AB - BACKGROUND: Angiography allows the definition of advanced, severe stages of coronary artery disease, but early atherosclerotic lesions, which do not lead to luminal stenosis, are not identified reliably. In contrast, intravascular ultrasound scanning allows the precise characterization and quantification of a wide range of atherosclerotic lesions, independent of the severity of luminal stenosis. METHODS: Three-dimensional (3-D) reconstruction of entire coronary segments is possible with the integration of sequential 2-dimensional tomographic images and allows volumetric analysis of coronary arteries. RESULTS: Automated systems able to recognize lumen and vessel borders and to display 3-D images are becoming available. CONCLUSION: These systems have the potential for on-line 3-D image reconstruction for clinical decision-making and fast routine volumetric analysis in research studies. This review describes 3-D intravascular ultrasound scanning acquisition, analysis, and processing, and the associated technical challenges. PMID- 12766736 TI - Use of complementary and alternative medical therapies in patients with cardiovascular disease. AB - BACKGROUND: Complementary and alternative medical (CAM) therapies are becoming increasingly popular, yet little information is available about the prevalence and patterns of CAM therapy use by patients with cardiovascular disease (CVD). METHODS: Interviewers administered telephone questionnaires to 107 patients randomly selected from a stratified cohort of 2487 eligible patients participating in a registry of patients with CVD. RESULTS: The current use of CAM therapies was reported by 64% of the patients surveyed. Nutritional supplements (40%) and megadose vitamins (35%) were the most frequently used preparations. Most CAM therapy users (65%) cited their underlying cardiac condition as the reason for taking such therapy. The most common sources of information about CAM were a friend or relative (43%) or the respondent's usual physician. However, although 80% of respondents claimed that they had discussed their use of CAM therapies with their physician, 58% of respondents taking a potentially toxic cardiovascular medication (digoxin, warfarin, sotalol, or amiodarone) were simultaneously taking an oral supplement. CONCLUSION: The use of CAM therapies was high in the cohort of patients surveyed. Physicians caring for patients with CVD need to inquire about CAM therapy use. Further scientific study should be performed to evaluate the potential benefits and risks of CAM therapies in this patient population. PMID- 12766737 TI - Plaque blush, branch location, and calcification are angiographic predictors of progression of mild to moderate coronary stenoses. AB - BACKGROUND: Angiographic predictors of plaque progression are weak and few: length, irregular surface, turbulence, low shear, and (in some studies) eccentricity and calcification. Having noted plaques that briefly retained dye after angiography, we interpreted these as plaques with a fissured surface or neovascularization and hypothesized that progression would be predicted by "plaque blush." METHODS: Plaques (<50% diameter stenosis) in 68 pairs of angiograms, 5.6 +/- 4.8 months apart, were reviewed by 2 blinded observers. The presence of plaque blush, calcification, clot (mobile defect), eccentricity, and a branch point location were compared between progressing (> or =20% stenosis increase) and nonprogressing plaques. RESULTS: Sixteen lesions in 15 patients progressed from 29% +/- 13% to 68% +/- 14% over a period of 8.1 +/- 7.9 months. Patients with and without progression were similar in sex, age, congestive heart disease risk factors, medications, interval between angiograms, clinical presentation, and initial stenosis severity. By logistic regression, plaque blush (BL) (P =.002), calcification (CA) (P =.024), and a branch (BR) point location (P =.001) predicted plaque progression. The odds ratio for plaque progression (ORp) was calculated as ORp = e(2.5 x BL + 1.8 x CA + 2.6 x BR). Using an ORp of 1/3, the model has 81% sensitivity and 77% specificity. A second analysis in which each progressive lesion was compared with proximal and distal lesions and with one in a different coronary artery yielded similar results. CONCLUSIONS: In mild to moderate coronary stenoses, studied retrospectively, plaque blush (a new sign) and a branch point location were strong predictors of plaque progression, whereas calcification was a weak predictor of progression. PMID- 12766738 TI - Clinical usefulness and prognostic value of elevated cardiac troponin I levels in acute pulmonary embolism. AB - BACKGROUND: Right ventricular myocardial ischemia and injury contribute to right ventricular dysfunction and failure during acute pulmonary embolism. The objective of this study was to evaluate the clinical usefulness of cardiac troponin I (cTnI) in the assessment of right ventricular involvement and short term prognosis in acute pulmonary embolism METHODS: Thirty-eight patients with acute pulmonary embolism were included in the study. Clinical characteristics, right ventricular involvement, and clinical outcome were compared in patients with elevated levels of serum cTnI versus patients with normal levels of serum cTnI. RESULTS: Among the study population (n = 38 patients), 18 patients (47%) had elevated cTnI levels (mean +/- SD 1.6 +/- 0.7 ng/mL, range 0.7-3.7 ng/mL, median, 1.4 ng/mL), and comprised the cTnI-positive group. In the other 20 patients, the serum cTnI levels were normal (< or =0.4 ng/mL), and they comprised the cTnI-negative group. In the cTnI-positive group (n = 18 patients), 12 patients (67%) had right ventricular dilatation/hypokinesia, compared with 3 patients (15%) in the cTnI-negative group (n = 20 patients, P =.004). Right ventricular systolic pressure was significantly higher in the cTnI-positive group (51 +/- 8 mm Hg vs 40 +/- 9 mm Hg, P =.002). Cardiogenic shock developed in a significantly higher number of patients with elevated serum cTnI levels (33% vs 5%, P =.01). In patients with elevated cTnI levels, the odds ratio for development of cardiogenic shock was 8.8 (95% CI 2.5-21). CONCLUSIONS: Patients with acute pulmonary embolism with elevated serum cTnI levels are at a higher risk for the development of right ventricular dysfunction and cardiogenic shock. Serum cTnI has a role in risk stratification and short-term prognostication in patients with acute pulmonary embolism. PMID- 12766739 TI - Time trends in long-term mortality after out-of-hospital cardiac arrest, 1980 to 1998, and predictors for death. AB - BACKGROUND: We studied time trends in long-term survival after out-of-hospital cardiac arrest (OHCA) for patient characteristics and described predictors for death after discharge. Because long-term prognosis among patients with coronary heart disease has improved in the last decades, we hypothesized that the prognosis after OHCA would improve with time. METHODS: We analyzed data that were prospectively collected from all patients discharged from the hospital after OHCA in the community of Goteborg, Sweden, from 1980 to 1998 and divided the data into 2 time periods, 1980 to 1991 and 1991 to1998, with an equal number of patients. RESULTS: A total of 430 patients were included in the survey. Age, sex proportions, cardiovascular comorbidity, resuscitation factors, and inhospital complications did not change with time. A diagnosis of a precipitating myocardial infarction was more common during period 1 (66% vs 54%). The prescription of aspirin (22% vs 52%), angiotensin-converting enzyme inhibitors (7% vs 29%), anticoagulants (13% vs 27%), and lipid-lowering agents (0% vs 6%) at discharge increased during period 2. Long-term survival did not improve with time; the 5 year mortality rates were 53% in period 1 and 52% in period 2. Independent predictors of an increased risk of death included age (risk ratio [RR] 1.06, 95% CI 1.05-1.08), history of myocardial infarction (RR 2.02, 95% CI 1.51-2.72), history of smoking (RR 1.77, 95% CI 1.29-2.44), and worse cerebral performance at discharge (RR 1.71, 95% CI 1.44-2.02). The prescription of beta-blockers at discharge was independently predictive of decreased risk of death (RR 0.63, 95% CI 0.46-0.85). CONCLUSION: The long-term survival rate after OHCA did not change. Baseline characteristics remained generally unchanged, but the drugs prescribed at discharge changed in several aspects. Age, a history of myocardial infarction, a history of smoking, cerebral performance category at discharge, and the prescription of beta-blockers were independent predictors of outcome. PMID- 12766740 TI - Insulin sensitivity in young women with vasovagal syncope. AB - BACKGROUND: Insulin, in addition to its known metabolic effects, has sympatho excitatory and vasodilatory actions on muscular blood vessels. The goal of this study was to evaluate insulin sensitivity in young women with vasovagal syncope and positive tilt test results (HUT+) and to compare it with that in patients with negative tilt test results (HUT-) and in control subjects without a history of syncope. METHODS: Different indices of insulin sensitivity were obtained by an oral glucose tolerance test (OGTT) in 13 young women with syncope and HUT+ (age 26.8 +/- 9.1 years, body mass index 20.4 +/- 2.1), 8 patients with HUT- (age 26 +/- 5.6 years, body mass index 21.9 +/- 2.4), and 13 control subjects without syncope and HUT- (age 28.9 +/- 8.8 years, body mass index 23.1 +/- 1.7). The following parameters were assessed: fasting glucose and insulin levels (G(0), I(0)); G(0)/I(0) ratio; G(0) x I(0); areas under the curve for glucose and insulin; homeostatic model assessment (HOMA); quantitative insulin sensitivity check index (QUICKI); and composite whole-body insulin sensitivity index (ISI). RESULTS: G(0) and I(0) values were significantly lower in patients with HUT+ than in control subjects (G(0) 4.9 vs 81.9, P <.05, I(0) 4.7 vs 9.1, P <.005). All the fasting values-based indices (ie, HOMA 0.9 vs 1.9, P <.005) and the ISI (12.8 vs 7.1, P =.01) differed significantly in both groups. None of the parameters showed significant differences between patients with HUT- and control subjects. Sixty one percent of patients with HUT+ had a vasovagal reaction during OGTT. CONCLUSIONS: Young women with vasovagal syncope and HUT+ have a greater insulin sensitivity. They have a propensity to reproduce symptoms during the OGTT. This hypersensitivity could be one of the predisposing factors for vasovagal episodes. PMID- 12766741 TI - Effect of an implantable cardioverter defibrillator with atrial detection and shock therapies on patient-perceived, health-related quality of life. AB - BACKGROUND: This study used a device (DDD implantable cardioverter defibrillator [ICD]) capable of delivering pacing and shock therapies to restore normal sinus rhythm in patients with atrial tachycardias or atrial fibrillation (AF). The purpose of this study was to assess the effect of the device on patient perceived, health-related quality of life (QOL). METHODS: The DDD ICD was implanted in 267 patients with drug refractory, symptomatic AF from 45 centers across Europe, the United States, and Canada. Patients completed self-reported, validated QOL assessments at baseline and at 3- and 6-month follow-up visits (The Medical Outcomes Short Form 36 [SF-36] and the Symptom Checklist [SCL]). RESULTS: The mean age of the study group was 62 +/- 12 years, and 73% of the patients were male. A total of 150 patients completed SF-36 assessments, and 138 patients completed SCL assessments at all 3 times. Baseline scores were more impaired (P <.05) on most SF-36 scales compared with norms for a general population, but were similar to a comparison group of patients with AF who were referred to tertiary care centers. The role-physical, physical functioning, vitality, mental health, and social functioning scales all improved significantly with time (all P <.04). Similarly, symptom frequency and severity (SCL) also improved significantly from baseline to 6 months (both P <.01). Shock therapy was delivered in 86 of the 150 patients (57%) with complete SF-36 evaluations. There was no evidence that receiving shocks decreased the relative improvement in QOL associated with implantation of the device. CONCLUSIONS: In a 6-month period, QOL improves after implantation of a DDD ICD with atrial shock and pacing therapies. These improvements were not attenuated by receipt of shocks. PMID- 12766742 TI - Comparison of acute changes in left ventricular volume, systolic and diastolic functions, and intraventricular synchronicity after biventricular and right ventricular pacing for heart failure. AB - BACKGROUND: Biventricular pacing (BiV) therapy has recently been shown to improve systolic function and cause reverse remodeling in patients with advanced heart failure with electromechanical delay. In these patients, the benefit of right ventricular (RV)-based pacing was controversial. We compared the acute changes in systolic and diastolic function, left ventricular (LV) volume, and intraventricular synchronicity in BiV pacing, RV pacing, and without pacing (No) by means of echocardiography and tissue Doppler imaging (TDI). METHODS: TDI was performed in 33 patients with heart failure after undergoing pacemaker implantation, when the device was randomized to BiV, RV, and no pacing modes. RESULTS: Systolic function was only improved during BiV pacing, but not during RV pacing. This included ejection fraction (No vs RV vs BiV = 24% +/- 12% vs 25% +/- 10% vs 30% +/- 14%, P =.02 vs No), +dp/dt (P =.01), myocardial performance index (P =.01), and isovolumic contraction time (P =.03). Mitral regurgitation was only reduced during BiV pacing (P =.02). LV early diastolic function was depressed in both RV and BiV pacing, as detected by transmitral flow (97 +/- 34 vs 80 +/- 34 vs 82 +/- 32 cm/s, both P < or =.005) and TDI (mean myocardial early diastolic velocity of 6 basal segments, 3.3 +/- 1.7 vs 2.6 +/- 1.0 vs 2.6 +/- 1.0 cm/s, both P =.01). The LV end-diastolic (187 +/- 86 vs 177 +/- 84 vs 166 +/- 79, P =.003) and end-systolic (146 +/- 77 vs 138 +/- 79 vs 122 +/- 69, P =.003) volumes were only decreased during BiV pacing. For systolic synchronicity, a significant delay in peak systolic contraction in the lateral over the septal wall (171 +/- 37 vs 217 +/- 46 ms, P =.004) was revealed by TDI when there was no pacing. This was abolished by BiV pacing, in which septal contraction was delayed (195 +/- 38 vs 201 +/- 53 ms, P = not significant). However, RV pacing restored the lateral wall delay, and systolic asynchrony reappeared (190 +/- 40 vs 227 +/- 56 ms, P =.01). Diastolic asynchrony between the septal and lateral walls was not evident in these patients and was not affected by either pacing mode. CONCLUSION: Only BiV pacing, but not RV pacing, improves systolic function, and reduces mitral regurgitation and LV volumes in patients with heart failure and electromechanical delay. This is attributed to the improvement of systolic synchronicity. Diastolic synchronicity was unaffected, whereas early diastolic function could be jeopardized, by either pacing mode. PMID- 12766743 TI - Comparison of rotational atherectomy with conventional balloon angioplasty in the prevention of restenosis of small coronary arteries: results of the Dilatation vs Ablation Revascularization Trial Targeting Restenosis (DART). AB - BACKGROUND: The optimum treatment of obstructive coronary disease in small (<3.0 mm diameter) arteries remains unknown. Rotational atherectomy is an approved treatment that might reduce the vascular injury during percutaneous coronary intervention compared with angioplasty. We report on a multicenter, randomized, blinded end point trial comparing rotational atherectomy with balloon angioplasty in the prevention of restenosis of obstructed small coronary arteries. METHODS: A total of 446 patients with myocardial ischemia associated with an angiographic stenosis in a native coronary artery 2 to 3 mm in diameter and < or =20 mm in length without severe calcification were randomly assigned to receive rotational atherectomy (n = 227) or balloon angioplasty (n = 219). The primary end point was target vessel failure at 12 months (defined as the composite of death, Q-wave myocardial infarction, and clinically driven repeat revascularization of the target vessel). RESULTS: The mean reference vessel diameter was 2.46 +/- 0.40 mm, the mean lesion length was 9.97 +/- 5.59 mm, and the prevalence of diabetes mellitus was 32%. Acute procedural success (91.6% for rotational atherectomy, 94.1% for balloon angioplasty, P =.36) and target vessel failure at 12 months were not significantly different (30.5% vs 31.2%, P =.98). At 8 months, there were no significant differences in minimum lumen diameter (1.28 +/- 0.63 mm vs 1.19 +/- 0.54 mm, P =.26), percent diameter stenosis (28% +/- 12% vs 29% +/- 15%, P =.59), binary restenosis rate (50.5% vs 50.5%, P = 1.0), or late loss index (0.57 vs 0.62, P =.7). No Q-wave myocardial infarctions occurred in either arm of the study, and non-Q-wave myocardial infarctions (defined as creatine kinase level >2 times normal with an elevated creatine kinase-myocardial band isoenzyme level) occurred in 2.2% and 1.4% of the patients in the rotational atherectomy and balloon angioplasty groups, respectively (P =.72). CONCLUSION: Rotational atherectomy was found to be safe in the treatment of obstructed small arteries, but lower rates of target vessel failure were not achieved compared with balloon angioplasty. Because the acute gain and loss index ratios of the 2 treatments were similar, there was no evident beneficial antirestenosis mechanism seen for rotational atherectomy. PMID- 12766744 TI - Outcomes of primary coronary angioplasty and angioplasty after initial thrombolysis in the treatment of 374 consecutive patients with acute myocardial infarction. AB - BACKGROUND: In patients with acute myocardial infarction (MI), the efficacy of thrombolysis is low. Angioplasty after failed thrombolysis (rescue percutaneous coronary angioplasty [PTCA]) has been associated with an increase in the incidence of inhospital complications. It has been proposed that these complications result from the procedure itself. Thus, the aim of this study was to compare the efficacy, inhospital complications, and mortality rate of patients with MI who are treated with primary PTCA and PTCA after initial thrombolysis (rescue or immediate rescue) in an experienced clinical center specializing in percutaneous coronary interventions. METHODS AND RESULTS: The study group consisted of consecutive patients with MI treated with primary PTCA (n = 195) or PTCA after initial thrombolysis (n = 179). The study was performed in a referral center with a 24-hour catheter-laboratory service. The success rate of the procedure was 90.5% and 88.2% in the PTCA after initial thrombolysis group and primary PTCA group, respectively. The groups did not differ in the frequency of reocclusion, emergency surgical revascularization (coronary artery bypass grafting), or stroke. In patients without cardiogenic shock, the inhospital mortality rates were 3.2% and 0.6% in the rescue and immediate rescue group and primary PTCA group, respectively (not significant). In a subgroup of patients with cardiogenic shock, the mortality rate was 36.0% in the initial thrombolysis PTCA group and 30.8% in the primary PTCA group. However, after successful PTCA in this subgroup, the mortality rate dropped to 18% and 10%, respectively. CONCLUSIONS: After initial thrombolysis, PTCA is safe, effective, and likely to restore grade 3 Thrombolysis In Myocardial Infarction flow in about 90% of patients. When available, immediate rescue PTCA should be performed in all patients, including patients with cardiogenic shock. PMID- 12766745 TI - Primary angioplasty with routine stenting compared with thrombolytic therapy in elderly patients with acute myocardial infarction. AB - BACKGROUND: Prior studies have yielded conflicting data on the advantage of primary angioplasty compared with thrombolysis in elderly patients with acute myocardial infarction (AMI). These studies, however, were performed before the contemporary widespread use of intracoronary stents and glycoprotien IIb/IIIa antagonists. METHODS: We prospectively compared the outcome of 130 consecutive elderly patients (aged > or =70 years) with ST-elevation AMI who were admitted to 2 similar neighboring medical centers. Patients were assigned to receive either thrombolytic therapy with accelerated tissue-type plasminogen activator (center I) or primary angioplasty with routine stenting (center II). RESULTS: Of the patients assigned to receive primary angioplasty, 91% underwent stenting. At 6 months, patients treated with primary angioplasty, compared with those treated with thrombolytic therapy, had a lower incidence of reinfarction (2% vs 14%, P =.053) and revascularization for recurrent ischemia (9% vs 61%, P <.001) and a significant reduction in the prespecified combined end point of death, reinfarction, or revascularization for recurrent ischemia (29% vs 93%, P <.01). Primary angioplasty remained an independent predictor of the triple combined end point after controlling for potential covariables (relative risk 0.63, 95% CI 0.38-0.84). Major bleeding complications were also significantly reduced in the primary angioplasty group (0% vs 17%, P =.03). CONCLUSIONS: Compared with thrombolysis, primary angioplasty with routine stenting in elderly patients with AMI is associated with better clinical outcomes and a lower risk of bleeding complications. PMID- 12766746 TI - Clinical and angiographic efficacy of a self-expanding nitinol stent in saphenous vein graft atherosclerotic disease: the Stent Comparative Restenosis (SCORES) Saphenous Vein Graft Registry. AB - BACKGROUND: The Stent Comparative Restenosis (SCORES) Saphenous Vein Graft (SVG) Registry was a multicenter, prospective registry designed to evaluate the safety and efficacy of a self-expanding, nickel-titanium (nitinol) stent for de novo SVG lesions. METHODS: In all, 159 patients with de novo vein graft lesions > or =2.75 and < or =4.25 mm in diameter and <30 mm in length underwent stenting with the Radius self-expanding stent. The primary end point was target vessel failure (TVF) at 9 months, which was defined as a composite of procedural failure, death, myocardial infarction, or target vessel revascularization. RESULTS: Procedural success was achieved in 96.8% of patients, and the 30-day incidence of major adverse cardiac events was 2.5%. The binary rate of restenosis at 6 months was 28.6%. By 9 months, the rate of TVF was 24.5%, and the rate of major adverse cardiac events was 23.1%. The 9-month Kaplan-Meier survival rates for freedom from TVF and target lesion revascularization were 76.0% and 87.9%, respectively. No clinical or angiographic characteristic was predictive of restenosis. CONCLUSIONS: In de novo atherosclerotic SVG disease, the use of a self-expanding, nitinol stent was associated with high initial procedural success and favorable early and intermediate outcomes. Because few studies have examined the influence of stent composition and design in SVG disease, these findings not only show the safety and efficacy of this self-expanding stent in de novo SVG disease, but also merit further comparison with balloon-expandable stents. PMID- 12766747 TI - Beta-blockers reduce the incidence of clinical restenosis: prospective study of 4840 patients undergoing percutaneous coronary revascularization. AB - BACKGROUND: Restenosis after percutaneous transluminal coronary intervention (PCI) remains a serious complication in the treatment of coronary artery disease. Although beta-adrenergic receptor blockers (BBs) effectively reduce many cardiac events, no large prospective studies have examined the association of BBs with restenosis. METHODS: We prospectively evaluated the association of BBs (prescribed at hospital discharge) with clinical restenosis in 4840 patients who underwent stent placement (60%), balloon angioplasty (32%), or rotational atherectomy (8%). Clinical restenosis was defined as repeat target lesion revascularization or coronary artery bypass grafting within 6 months of PCI. Other end points included 9-month clinical restenosis, repeat target lesion PCI (only), long-term (5-year) target lesion repeat-PCI, and major adverse cardiac events (MACE). Multivariable regression adjusted the effect of BBs on clinical restenosis for 15 covariables. RESULTS: The average patient age was 63 years, 75% were men, and 37% received a BB prescription. The incidence of clinical restenosis was 12% overall and was lower among those prescribed a BB (10.0% for BB, 13.5% for none, adjusted odds ratio [OR] 0.76, P =.004). Other predictors of decreased restenosis included stent use, age, and smoking; predictors of increased restenosis included diabetes, atherectomy, and number of treated vessels. BBs also reduced 9-month clinical restenosis (10.3% vs 13.5%, OR 0.75, P =.004), MACE (16.5% vs 20.9%, OR 0.75, P <.001), 6-month target lesion restenosis (7.8% vs 10.2%, OR 0.75, P =.006), and 5-year target lesion restenosis (12.0% vs 14.0%, OR 0.83, P =.046). CONCLUSIONS: beta-Adrenergic receptor blockers prescribed after PCI reduced the risk of clinical restenosis, target lesion restenosis, and MACE in this cohort of 4840 patients. The mechanism by which beta blockers conferred a protective effect against restenosis remains to be determined. PMID- 12766748 TI - Cost-effectiveness of measuring fractional flow reserve to guide coronary interventions. AB - BACKGROUND: Most patients come to the catheterization laboratory without prior functional tests, which makes the cost-effective treatment of patients with intermediate coronary lesions a practical challenge. METHODS: We developed a decision model to compare the long-term costs and benefits of 3 strategies for treating patients with an intermediate coronary lesion and no prior functional study: 1) deferring the decision for percutaneous coronary intervention (PCI) to obtain a nuclear stress imaging study (NUC strategy); 2) measuring fractional flow reserve (FFR) at the time of angiography to help guide the decision for PCI (FFR strategy); and 3) stenting all intermediate lesions (STENT strategy). On the basis of the literature, we estimated that 40% of intermediate lesions would produce ischemia, 70% of patients treated with PCI and 30% of patients treated medically would be free of angina after 4 years, and the quality-of-life adjustment for living with angina was 0.9 (1.0 = perfect health). We estimated the cost of FFR to be 761 dollars, the cost of nuclear stress imaging to be 1093 dollars, and the cost of medical treatment for angina to be 1775 dollars per year. The extra cost of splitting the angiogram and PCI as dictated by the NUC strategy was 3886 dollars by use of hospital cost-accounting data. Sensitivity and threshold analyses were performed to determine which variables affected our results. RESULTS: The FFR strategy saved 1795 dollars per patient compared with the NUC strategy and 3830 dollars compared with the STENT strategy. Quality adjusted life expectancy was similar among the 3 strategies (NUC-FFR = 0.8 quality-adjusted days, FFR-STENT = 6 quality-adjusted life days). Compared with the FFR strategy, the NUC strategy was expensive (>800,000 dollars per quality adjusted life year gained). Both screening strategies were superior to (less cost, better outcomes) the STENT strategy. Sensitivity analysis indicated that the NUC strategy would only become attractive (<50,000 dollars/quality-adjusted life years compared with FFR) if the specificity of nuclear stress imaging was >25% better than FFR. Our results were not altered significantly by changing the other assumptions. CONCLUSION: In patients with an intermediate coronary lesion and no prior functional study, measuring FFR to guide the decision to perform PCI may lead to significant cost savings compared with performing nuclear stress imaging or with simply stenting lesions in all patients. PMID- 12766749 TI - Preventing heart disease by controlling hypertension: impact of hypertensive subtype, stage, age, and sex. AB - BACKGROUND: Hypertension is related to significant morbidity and mortality rates from coronary heart disease (CHD). This report examines the relative and absolute impact on risk for CHD by controlling hypertension to high normal and optimal levels. METHODS: Among all subjects with untreated or inadequately treated hypertension in the National Health and Nutrition Examination Survey (NHANES) III who were 30 to 74 years of age and without prior CHD, the 10-year risk of CHD was calculated. With the use of sampling weights, the number of CHD events by age group, hypertension subtype (isolated diastolic hypertension [IDH], systolic diastolic hypertension [SDH], and isolated systolic hypertension [ISH]), and stage of hypertension was estimated. Risk was recalculated and the number of events reestimated, assuming a reduction in blood pressure (BP) to high normal and optimal levels. The number and proportion (population-attributable risk, or PAR%) of events that could be prevented were determined from the differences in events and risk between uncontrolled and controlled BP levels. Derived from this was the number of persons needing treatment per CHD event prevented. RESULTS: Control of hypertension to high normal levels could prevent approximately one fifth (PAR = 19%) of CHD events in men and one third (PAR = 31%) of CHD events in women, whereas control to optimal levels may prevent 37% and 56% of CHD events, respectively (P <.01 for differences between men and women). Of CHD events that could be prevented, the greatest proportion occurred from controlling BP among older persons, men, and those with stage 1 hypertension (vs stages 2 and 3) or with ISH (vs IDH or SDH). The number of persons with hypertension needing treatment to prevent one CHD event ranged from 20.5 in men to 38.6 in women when controlled to high normal BP and 10.7 in men and 21.3 in women when controlled to optimal BP. CONCLUSIONS: The greatest impact from control of hypertension occurs in older persons, men, and those with ISH, whereas the greatest PAR% occurred in women. Optimal control of BP could prevent more than one third of CHD events in men and more than half of events in women. Greater efforts to control hypertension in these populations may have a substantial impact in preventing CHD events. PMID- 12766750 TI - Imaging and diagnostic testing: diastolic dysfunction after coronary artery bypass grafting: a frequent finding of clinical significance not influenced by intravenous calcium. AB - BACKGROUND: Diastolic dysfunction is common immediately after coronary artery bypass surgery (CABG). The duration of this phenomenon is unknown. Intravenous calcium is frequently administered during separation from cardiopulmonary bypass (CPB). We sought to determine whether intravenous calcium influences perioperative diastolic function and whether diastolic dysfunction persists into the postoperative period. METHODS AND RESULTS: Patients undergoing first-time elective CABG (n = 29) were randomly assigned to receive intravenous calcium chloride (n = 13) or placebo (n = 16) during separation from CPB. Diastolic function was assessed by the pressure-area relation with transesophageal echocardiography and pulmonary capillary wedge pressure (PCWP) measured simultaneously. Left ventricular end-diastolic area (LVEDA) and Doppler indexes were measured at comparable PCWP (within 2 mm Hg) at baseline, after separation from CPB, after sternal closure, and 3 hours after surgery. After CABG, both groups had a significant decrease in LVEDA and mitral E-wave deceleration time that persisted at 3 hours. Because there were no significant differences between the calcium and control groups at any time point, the data for the entire study cohort was analyzed. The LVEDA decreased (stiffness increased) progressively from 16.9 +/- 3.4 cm2 at baseline to 15.8 +/- 2.9 cm2 after CPB, 14.9 +/- 2.5 cm2 after sternal closure, and 14.3 +/- 3.1 cm2 at 3 hours after surgery (P <.0001). The mitral E-wave deceleration time measured at the same time points was 168 +/- 47 ms, 136 +/- 25 ms, 137 +/- 36 ms, and 111 +/- 44 ms (P =.0001). CONCLUSIONS: An increase in left ventricular diastolic chamber stiffness is nearly universal after CABG, and it persists for at least 3 hours after surgery. An intravenous bolus of calcium chloride given during separation from CPB has no measurable negative effect on diastolic function. In the setting of increased chamber stiffness, the PCWP alone does not adequately reflect the volume status and effective preload of the left ventricle. PMID- 12766751 TI - Effect of dietary intervention and lipid-lowering treatment on brachial vasoreactivity in patients with ischemic heart disease and hypercholesterolemia. AB - BACKGROUND: The "Mediterranean" diet and statin treatment have both independently been shown to improve survival and reduce the risk of cardiovascular events in patients with ischemic heart disease (IHD), but no studies have evaluated the effect of this combination on endothelial function. We therefore sought to evaluate the effect of the combination dietary intervention and lipid-lowering treatment on brachial vasoreactivity. METHODS: A total of 131 consecutive patients with documented IHD and a serum cholesterol level > or =5 mmol/L (193 mg/dL) were randomized to receive Mediterranean dietary advice (n = 68) or no specific dietary advice (n = 63). Endothelial function was assessed at baseline and after 12 months with noninvasive ultrasound scanning vessel-wall tracking of brachial artery flow-mediated vasodilatation (FMD). All patients started statin treatment with Fluvastatin (40 mg once daily) at baseline. RESULTS: A total of 115 patients completed the study. At baseline, FMD was 4.30% +/- 4.89% in the control group versus 4.32% +/- 6.15% in the intervention group (P = not significant). After 12 months of follow-up, FMD was significantly higher in the intervention group (control group 5.72% +/- 4.87% vs intervention group 8.62% +/- 6.60%, P <.01). This was accompanied by a larger intake of fatty fish and a significant decrease in triglyceride levels. In multivariate analysis, randomization status was a significant predictor of FMD after adjustment for classic cardiovascular risk factors and vessel size (P =.02; beta = -2.66 [-4.91; -0.41]). CONCLUSION: Dietary intervention with the Mediterranean diet and statin treatment improve FMD in the brachial artery in patients with IHD and hypercholesterolemia to a greater degree than statin treatment alone. PMID- 12766752 TI - Mechanisms of exercise training in patients with heart failure. AB - BACKGROUND: The reduction of exercise capacity because of fatigue and dyspnea in patients with heart failure can be improved with exercise training. We sought to examine the mechanisms of exercise training as an adjunctive treatment strategy for patients with heart failure. METHODS: We reviewed the published data on the possible mechanisms of effect of exercise training in heart failure. RESULTS: Symptoms of heart failure may be explained on the basis of abnormal skeletal muscle perfusion and structure and endothelial function. Exercise training has been shown to engender changes in muscle structure and biochemistry and vascular function, although effects on cardiac function have not been detected uniformly and may require longer training periods. CONCLUSIONS: A suitable, long-term program of exercise training may reverse unfavorable interactions among the heart, vessels, and skeletal muscles. These improvements may be preserved with an ongoing maintenance program. PMID- 12766753 TI - Differential effects of exercise training in men and women with chronic heart failure. AB - BACKGROUND: Abnormalities of myosin heavy chain (MHC) isoforms, enzyme activity, and capillarity contribute to the exercise intolerance that is characteristic of patients with heart failure. To what extent these changes can be reversed with exercise training and whether differences exist in the responses of men and women remains uncertain. We described and compared the effects of exercise training on exercise capacity and skeletal muscle histochemistry in men and women with chronic heart failure. METHODS: Fifteen patients (10 male) undergoing standard medical therapy completed a 14- to 24-week exercise training program. Peak oxygen consumption, MHC isoforms, capillary density, and selected metabolic enzymes were assessed before and after training. RESULTS: Peak oxygen consumption was improved 14% (P <.05); however, this increase was mostly because of the improvement observed in men versus women (+20% versus +2%, respectively, P <.01). At baseline, MHC I content was lower in men than in women (33% +/- 3% vs 49.6% +/- 5.5%, P <.05). MHC I improved with training in men, to 45.6% +/- 4.5% (+38%, P <.05), versus women (-3%, P =.82), and the increase in men tended (P =.12) to be significant when compared with that in women. There were no significant changes in capillary density or muscle enzyme activity with training in the group as a whole or in men and women separately. CONCLUSION: Among patients with chronic heart failure, improvements in peak exercise capacity may be more pronounced in men than in women. This difference in response of functional capacity to training paralleled differences observed between men and women for changes in MHC I isoforms. PMID- 12766754 TI - Antibiotics in primary prevention of myocardial infarction among elderly patients with hypertension. AB - BACKGROUND: Given the premise that certain bacteria (such as Chlamydia pneumoniae) may play a role in the etiology of atherosclerosis, subjects treated with antibiotics that have antibacterial activity against C pneumoniae may be at lower risk for the development of an acute myocardial infarction (MI) than untreated subjects. METHODS: A case-control design, nested within a cohort of 29,937 elderly subjects in whom antihypertensive therapy was initiated (1982 1995) was used, in which each subject who was hospitalized with a primary discharge diagnosis of MI between 1987 and 1995 (n = 1047) was matched on calendar time to 5 randomly selected control subjects for exposure contrasts. Conditional logistic regression analyses were conducted to adjust for predisposing factors for MI. RESULTS: Although no clear consistent effect of antibiotics use was found in relation to MI, a trend was observed for a decreased risk of acute MI in patients receiving a prescription for antichlamydial antibiotics in the preceding 3 months (odds ratio 0.68, 95% CI 0.46-1.00). Antibiotics without antichlamydial activity showed no benefit in MI risk. CONCLUSION: The beneficial effect of certain antichlamydial antibiotics in reducing the risk of MI cannot be excluded on the basis of this representative cohort of elderly patients in a routine clinical care setting. Larger prospective studies are required to confirm the usefulness of antibiotics in the primary prevention of MI. PMID- 12766755 TI - Relation of QT interval and QT dispersion to regression of echocardiographic and electrocardiographic left ventricular hypertrophy in hypertensive patients: the Losartan Intervention For Endpoint Reduction (LIFE) study. AB - BACKGROUND: In hypertensive patients, both echocardiographic and electrocardiographic left ventricular hypertrophy (LVH) increase the risk of sudden death, possibly in part because of LVH-induced proarrhythmic repolarization changes. Experimentally, regression of LVH normalizes ventricular electrophysiology. METHODS: To assess the relation of regression of LVH to changes in electrocardiographic measures of ventricular repolarization, we studied 317 hypertensive (61.2% men, mean age 65 +/- 7 years) participants in the Losartan Intervention For Endpoint Reduction (LIFE) study with electrocardiographic evidence of LVH, at study baseline, and after 1 year of blood pressure-lowering treatment with losartan or atenolol and hydrochlorothiatzide as the first adjunct therapy if needed to reach target blood pressure of 140/90 mm Hg. As indexes of LVH, we used echocardiographically determined LV mass as well as the Sokolow-Lyon and Cornell voltages from the electrocardiogram. QT interval duration and QT dispersion from the 12-lead electrocardiogram were used as ventricular repolarization measures. RESULTS: By using tertiles of LV mass change and adjusting for the difference in treatment (losartan or atenolol), shortening of the rate-adjusted QT intervals as well as reduction in QT(apex) dispersion were observed in the tertile showing the greatest decrease in LV mass but not in the tertile without substantial changes in LV mass despite a significant reduction in blood pressure. Similar results were obtained with the use of Sokolow-Lyon and Cornell voltage change tertiles. CONCLUSIONS: In hypertensive patients with electrocardiographic evidence of LVH, regression of echocardiographically determined LV mass and electrocardiographic indexes of LVH may partially reverse the LVH-induced proarrhythmic repolarization changes. This may have a beneficial impact on the increased incidence of sudden death in these patients. PMID- 12766756 TI - Prognostic importance of the oxidized product of catecholamines, adrenolutin, in patients with severe heart failure. AB - OBJECTIVES: The purpose of this study was to assess whether adrenolutin, the inert product of the highly reactive molecules aminochromes, is increased in severe chronic heart failure and whether it is associated with a poor prognosis. BACKGROUND: Experimental evidence suggests that oxidative products of catecholamines, aminochromes, are more cardiotoxic than unoxidized catecholamines and may be increased in heart failure. METHODS: Adrenolutin was measured at baseline and at 1 and 3 months in 263 patients with chronic New York Heart Association class III or IV heart failure and a left ventricular ejection fraction of 22% +/- 7%. Adrenolutin levels were compared with normal levels, and their relation to prognosis was evaluated. RESULTS: Baseline adrenolutin was increased (55 +/- 90 pg/mL vs 8.4 +/- 9.1 pg/mL for control, P <.02) and remained increased at 1 month (49 +/- 65 pg/mL). During a mean follow-up of 309 +/- 148 days (22-609 days), 57 patients died. Baseline adrenolutin levels correlated with mortality rates by univariate and multivariate analyses (relative risk 1.06, 95% CI 1.01-1.10 for each 17.9-pg/mL rise, P =.032). Left ventricular ejection fraction (P =.013) and New York Heart Association class (P =.009) were the only other variables associated with survival. Age, sex, plasma creatinine, plasma N terminal atrial natriuretic peptide, and plasma norepinephrine levels were not retained in our model. Adrenolutin levels 1 month after random assignment were not significantly correlated with total mortality rate (P =.061) but were correlated with mortality rate from low output (relative risk 1.14, 95% CI 1.06 1.22, P =.002). CONCLUSIONS: Plasma adrenolutin is increased in patients with heart failure and correlates with a poor prognosis independent of other important predictors of survival. This finding has potentially important pathophysiologic, prognostic, and therapeutic implications. PMID- 12766757 TI - A conformational switch between transcriptional repression and replication initiation in the RepA dimerization domain. AB - Plasmids are natural vectors for gene transfer. In Gram-negative bacteria, plasmid DNA replication is triggered when monomers of an initiator protein (Rep) bind to direct repeats at the origin sequence. Rep dimers, which are inactive as initiators, bind to an inverse repeat operator, repressing transcription of the rep gene. Rep proteins are composed of N-terminal dimerization and C-terminal DNA binding domains. Activation of Rep is coupled to dimer dissociation, converting the dimerization domain into a second origin-binding module. Although the structure of the monomeric F plasmid initiator (mRepE) has been determined, the molecular nature of Rep activation remains unknown. Here we report the crystal structure of the dimeric N-terminal domain of the pPS10 plasmid initiator (dRepA). dRepA has a winged-helix fold, as does its homologous domain in mRepE. However, dimerization transforms an interdomain loop and beta-strand (monomeric RepE) into an alpha-helix (dimeric RepA). dRepA resemble the C terminus of eukaryotic and archaeal Cdc6, giving clues to the phylogeny of DNA replication initiators. PMID- 12766758 TI - The ABCs of granule-mediated cytotoxicity: new weapons in the arsenal. AB - Granule exocytosis is the main pathway for the immune elimination of virus infected cells and tumour cells by cytotoxic T lymphocytes and natural killer cells. After target-cell recognition, release of the cytotoxic granule contents into the immunological synapse formed between the killer cell and its target induces apoptosis. The granules contain two membrane-perturbing proteins, perforin and granulysin, and a family of serine proteases known as granzymes, complexed with the proteoglycan serglycin. In this review, I discuss recent insights into the mechanisms of granule-mediated cytotoxicity, focusing on how granzymes A, B and C and granulysin activate cell death through caspase independent pathways. PMID- 12766759 TI - NODs: intracellular proteins involved in inflammation and apoptosis. AB - NOD (nucleotide-binding oligomerization domain) proteins are members of a family that includes the apoptosis regulator APAF1 (apoptotic protease activating factor 1), mammalian NOD-LRR (leucine-rich repeat) proteins and plant disease-resistance gene products. Several NOD proteins have been implicated in the induction of nuclear factor-kappaB (NF-kappaB) activity and in the activation of caspases. Two members of the NOD family, NOD1 and NOD2, mediate the recognition of specific bacterial components. Notably, genetic variation in the genes encoding the NOD proteins NOD2, cryopyrin and CIITA (MHC class II transactivator) in humans and Naip5 (neuronal apoptosis inhibitory protein 5) in mice is associated with inflammatory disease or increased susceptibility to bacterial infections. Mammalian NOD proteins seem to function as cytosolic sensors for the induction of apoptosis, as well as for innate recognition of microorganisms and regulation of inflammatory responses. PMID- 12766760 TI - Negative selection--clearing out the bad apples from the T-cell repertoire. AB - Dead cells are a prominent feature of the thymic landscape as only 5% of developing thymocytes are exported as mature T cells. The remaining thymocytes die by one of two mechanisms; most thymocytes die because they are not positively selected and do not receive a survival signal, whereas a minority of thymocytes undergo T-cell receptor (TCR)-mediated apoptosis, a process known as negative selection. Negative selection is extremely important for establishing a functional immune system, as it provides an efficient mechanism for ridding the T cell repertoire of self-reactive and potentially autoimmune lymphocytes. This review discusses several cellular and molecular aspects of negative selection. PMID- 12766761 TI - Apoptosis as an HIV strategy to escape immune attack. AB - Viruses have evolved numerous mechanisms to evade the host immune system and one of the strategies developed by HIV is to activate apoptotic programmes that destroy immune effectors. Not only does the HIV genome encode pro-apoptotic proteins, which kill both infected and uninfected lymphocytes through either members of the tumour-necrosis factor family or the mitochondrial pathway, but it also creates a state of chronic immune activation that is responsible for the exacerbation of physiological mechanisms of clonal deletion. This review discusses the molecular mechanisms by which HIV manipulates the apoptotic machinery to its advantage, assesses the functional consequences of this process and evaluates how new therapeutics might counteract this strategy. PMID- 12766762 TI - To respond or not to respond: T cells in allergic asthma. AB - The incidence of allergic asthma has almost doubled in the past two decades. Numerous epidemiological studies have linked the recent surge in atopic disease with decreased exposure to infections in early childhood as a result of a more westernized lifestyle. However, a clear mechanistic explanation for how this might occur is still lacking. An answer might lie in the presently unfolding story of various regulatory T-cell populations that can limit adaptive immune responses, including T helper 2 (T(H)2)-cell-mediated allergic airway disease. PMID- 12766763 TI - What does it take to make a natural killer? AB - We know how B and T cells develop, what they 'see' and the receptors they 'see with'. By contrast, and despite an unprecedented increase in the number of receptors and ligands known to regulate the activity of natural killer (NK) cells, we still have many questions regarding how these cells develop. Nevertheless, we are beginning to understand the transcriptional programmes of NK cell maturation and the role of the effector functions of NK cells in the regulation of immune responses. An improved knowledge of NK-cell development in mice and humans might be useful to harness the power of these natural killers in the clinic to fight autoimmune diseases, infection and cancer. PMID- 12766764 TI - Re-evaluating the role of heat-shock protein-peptide interactions in tumour immunity. AB - Early investigations into the immune surveillance of chemically-induced sarcomas led to two important concepts in tumour immunobiology: one, tumour rejection can be elicited by immune recognition of tumour antigens; and two, tumours express unique sets of antigens, which are known as tumour-specific antigens. The pioneering studies of Srivastava and colleagues led to the proposal that heat shock proteins (HSPs) function as ubiquitous tumour-specific antigens, with the specificity residing in a population of bound peptides that identify the tissue of origin of the HSP. However, recent findings, including new data on the cell biology of peptide generation and trafficking, have called into question the specificity of tumour rejection that is induced by HSPs. PMID- 12766765 TI - Enhanced T-cell immunogenicity of plasmid DNA vaccines boosted by recombinant modified vaccinia virus Ankara in humans. AB - In animals, effective immune responses against malignancies and against several infectious pathogens, including malaria, are mediated by T cells. Here we show that a heterologous prime-boost vaccination regime of DNA either intramuscularly or epidermally, followed by intradermal recombinant modified vaccinia virus Ankara (MVA), induces high frequencies of interferon (IFN)-gamma-secreting, antigen-specific T-cell responses in humans to a pre-erythrocytic malaria antigen, thrombospondin-related adhesion protein (TRAP). These responses are five to tenfold higher than the T-cell responses induced by the DNA vaccine or recombinant MVA vaccine alone, and produce partial protection manifest as delayed parasitemia after sporozoite challenge with a different strain of Plasmodium falciparum. Such heterologous prime-boost immunization approaches may provide a basis for preventative and therapeutic vaccination in humans. PMID- 12766767 TI - Hematopoietic stem cells engraft in mice with absolute efficiency. AB - The engraftment of murine hematopoietic stem cells (HSCs) into irradiated mice is thought to be an inefficient process, but has yet to be measured directly. We used two independent strategies to test their engraftment efficiency: one measured competition of unpurified donor bone marrow cells with recipient cells in murine hosts and the other tracked the engraftment of one highly purified stem cell injected per recipient. The results showed that stem cells engrafted with near absolute efficiency. Thus, inefficient engraftment cannot explain the low frequency of permanent reconstitutions observed with pure HSC fractions and instead suggests most initially engrafted cells fail to sustain self-renewal. PMID- 12766766 TI - The inhibitory function of B7 costimulators in T cell responses to foreign and self-antigens. AB - When antigen-presenting cells (APCs) encounter inflammatory stimuli, they up regulate their expression of B7. A small amount of B7 is also constitutively expressed on resting APCs, but its function is unclear. Here we show that initiation of T cell responses requires the expression of B7 on immunizing APCs, but the responses are much greater in the absence of basal B7 expression. Transfer of antigen-specific CD4+CD25+ cells reverses the increased responsiveness, and tolerance to a self-protein is broken by immunization in the absence of basal B7, thereby inducing autoimmunity. Similar loss of self tolerance is seen on depletion of CD25+ cells. Thus, constitutively expressed B7 costimulators function to suppress T cell activation and maintain self-tolerance, principally by sustaining a population of regulatory T cells. PMID- 12766768 TI - CpG directly induces T-bet expression and inhibits IgG1 and IgE switching in B cells. AB - CpG DNA has immunomodulatory effects, such as the suppression of allergic responses mediated by type II T cell help (T(H)2). Here we report that CpG, but not lipopolysaccharide (LPS), rapidly induces expression of T-bet mRNA in purified B cells. Up-regulation of T-bet by CpG is abrogated in mice deficient in Toll-like receptor 9 (TLR9) and MyD88, but remains intact in B cells deficient in STAT1 (signal transducer and activator of transcription 1). Interleukin 12 (IL 12) alone does not up-regulate T-bet mRNA, but greatly enhances CpG-induced T-bet expression. Furthermore, CpG inhibits immunoglobulin G1 (IgG1) and IgE switching induced by IL-4 and CD40 signaling in purified B cells, and this effect correlates with up-regulation of T-bet. Thus, CpG triggers anti-allergic immune responses by directly regulating T-bet expression via a signaling pathway in B cells that is dependent upon TLR9, independent of interferon-gamma (IFN-gamma) STAT1 and synergistic with IL-12. PMID- 12766770 TI - Fras1 deficiency results in cryptophthalmos, renal agenesis and blebbed phenotype in mice. AB - Loss of tight association between epidermis and dermis underlies several blistering disorders and is frequently caused by impaired function of extracellular matrix (ECM) proteins. Here we describe a new protein in mouse, Fras1, that is specifically detected in a linear fashion underlying the epidermis and the basal surface of other epithelia in embryos. Loss of Fras1 function results in the formation of subepidermal hemorrhagic blisters as well as unilateral or bilateral renal agenesis during mouse embryogenesis. Postnatally, homozygous Fras1 mutants have fusion of the eyelids and digits and unilateral renal agenesis or dysplasia. The defects observed in Fras1-/- mice phenocopy those of the existing bl (blebbed) mouse mutants, which have been considered a model for the human genetic disorder Fraser syndrome. We show that bl/bl homozygous embryos are devoid of Fras1 protein, consistent with the finding that Fras1 is mutated in these mice. In sum, our data suggest that perturbations in the composition of the extracellular space underlying epithelia could account for the onset of the blebbed phenotype in mouse and Fraser syndrome manifestation in human. PMID- 12766769 TI - Fraser syndrome and mouse blebbed phenotype caused by mutations in FRAS1/Fras1 encoding a putative extracellular matrix protein. AB - Fraser syndrome (OMIM 219000) is a multisystem malformation usually comprising cryptophthalmos, syndactyly and renal defects. Here we report autozygosity mapping and show that the locus FS1 at chromosome 4q21 is associated with Fraser syndrome, although the condition is genetically heterogeneous. Mutation analysis identified five frameshift mutations in FRAS1, which encodes one member of a family of novel proteins related to an extracellular matrix (ECM) blastocoelar protein found in sea urchin. The FRAS1 protein contains a series of N-terminal cysteine-rich repeat motifs previously implicated in BMP metabolism, suggesting that it has a role in both structure and signal propagation in the ECM. It has been speculated that Fraser syndrome is a human equivalent of the blebbed phenotype in the mouse, which has been associated with mutations in at least five loci including bl. As mapping data were consistent with homology of FRAS1 and bl, we screened DNA from bl/bl mice and identified a premature termination of mouse Fras1. Thus, the bl mouse is a model for Fraser syndrome in humans, a disorder caused by disrupted epithelial integrity in utero. PMID- 12766771 TI - Specification of dorsal telencephalic character by sequential Wnt and FGF signaling. AB - Dorsoventral patterning of the telencephalon is established early in forebrain development and underlies many of the regional subdivisions that are critical to the later organization of neural circuits in the cerebral cortex and basal ganglia. Sonic hedgehog (Shh) is involved in the generation of the ventral-most telencephalic cells, but the identity of the extrinsic signal(s) that induce dorsal character in telencephalic cells is not known. Here we show in chick embryos that sequential Wnt and fibroblast growth factor (FGF) signaling specifies cells of dorsal telencephalic character. PMID- 12766772 TI - MKP3 mediates the cellular response to FGF8 signalling in the vertebrate limb. AB - The mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) and phosphatidylinositol-3-OH kinase (PI3K)/Akt pathways are involved in the regulatory mechanisms of several cellular processes including proliferation, differentiation and apoptosis. Here we show that during chick, mouse and zebrafish limb/fin development, a known MAPK/ERK regulator, Mkp3, is induced in the mesenchyme by fibroblast growth factor 8 (FGF8) signalling, through the PI3K/Akt pathway. This correlates with a high level of phosphorylated ERK in the apical ectodermal ridge (AER), where Mkp3 expression is excluded. Conversely, phosphorylated Akt is detected only in the mesenchyme. Constitutively active Mek1, as well as the downregulation of Mkp3 by small interfering RNA (siRNA), induced apoptosis in the mesenchyme. This suggests that MKP3 has a key role in mediating the proliferative, anti-apoptotic signalling of AER-derived FGF8. PMID- 12766773 TI - Centrosome number is controlled by a centrosome-intrinsic block to reduplication. AB - The centrosome duplicates once in S phase. To determine whether there is a block in centrosome reduplication, we used a cell fusion assay to compare the duplication potential of unduplicated G1 centrosomes and recently duplicated G2 centrosomes. By fusing cells in different cell cycle stages, we found that G2 centrosomes were unable to reduplicate in a cellular environment that supports centrosome duplication. Furthermore, G2 cytoplasm did not inhibit centrosome duplication in fused cells, indicating that the block to reduplication is intrinsic to the centrosomes rather than the cytoplasm. To test the underlying mechanism, we created mononucleate G1 cells with two centrosomes by fusing cells with enucleated cytoplasts. Both centrosomes duplicated, indicating that the block is not controlled by centrosome:nucleus ratio. We also found that human primary cells have tight control over centrosome number during prolonged S-phase arrest and that this control is partially abrogated in transformed cells. This suggests a link between the control of centrosome duplication and maintenance of genomic stability. PMID- 12766774 TI - Dual phosphorylation controls Cdc25 phosphatases and mitotic entry. AB - Negative regulation of the Cdc25C protein phosphatase by phosphorylation on Ser 216, the 14-3-3-binding site, is an important regulatory mechanism used by cells to block mitotic entry under normal conditions and after DNA damage. During mitosis, Cdc25C is not phosphorylated on Ser 216 and ionizing radiation (IR) does not induce either phosphorylation of Ser 216, or binding to 14-3-3. Here, we show that Cdc25C is phosphorylated on Ser 214 during mitosis, which in turn prevents phosphorylation of Ser 216. Mutation of Ser 214 to Ala reconstitutes Ser 216 phosphorylation and 14-3-3 binding during mitosis. Introduction of exogenous Cdc25C(S214A) into HeLa cells depleted of endogenous Cdc25C results in a substantial delay to mitotic entry. This effect was fully reversed in a S214A/S216A double-mutant, implying that the inhibitory effect of S214A mutant was entirely dependent on Ser 216 phosphorylation. A similar regulatory mechanism may also apply to another mitotic phosphatase, Cdc25B, as well as mitotic phosphatases of other species, including Xenopus laevis. We propose that this pathway ensures that Cdc2 remains active once mitosis is initiated and is a key control mechanism for maintaining the proper order of cell-cycle transitions. PMID- 12766775 TI - Rheb is an essential regulator of S6K in controlling cell growth in Drosophila. AB - Understanding the mechanisms through which multicellular organisms regulate cell, organ and body growth is of relevance to developmental biology and to research on growth-related diseases such as cancer. Here we describe a new effector in growth control, the small GTPase Rheb (Ras homologue enriched in brain). Mutations in the Drosophila melanogaster Rheb gene were isolated as growth-inhibitors, whereas overexpression of Rheb promoted cell growth. Our genetic and biochemical analyses suggest that Rheb functions downstream of the tumour suppressors Tsc1 (tuberous sclerosis 1)-Tsc2 in the TOR (target of rapamycin) signalling pathway to control growth, and that a major effector of Rheb function is ribosomal S6 kinase (S6K). PMID- 12766776 TI - Rheb promotes cell growth as a component of the insulin/TOR signalling network. AB - Insulin signalling is a potent stimulator of cell growth and has been proposed to function, at least in part, through the conserved protein kinase TOR (target of rapamycin) [corrected]. Recent studies suggest that the tuberous sclerosis complex Tsc1-Tsc2 may couple insulin signalling to Tor activity [corrected]. However, the regulatory mechanism involved remains unclear, and additional components are most probably involved. In a screen for novel regulators of growth, we identified Rheb (Ras homologue enriched in brain), a member of the Ras superfamily of GTP-binding proteins. Increased levels of Rheb in Drosophila melanogaster promote cell growth and alter cell cycle kinetics in multiple tissues. In mitotic tissues, overexpression of Rheb accelerates passage through G1-S phase without affecting rates of cell division, whereas in endoreplicating tissues, Rheb increases DNA ploidy. Mutation of Rheb suspends larval growth and prevents progression from first to second instar. Genetic and biochemical tests indicate that Rheb functions in the insulin signalling pathway downstream of Tsc1 Tsc2 and upstream of TOR. Levels of rheb mRNA are rapidly induced in response to protein starvation, and overexpressed Rheb can drive cell growth in starved animals, suggesting a role for Rheb in the nutritional control of cell growth. PMID- 12766777 TI - Colocalization of multiple DNA double-strand breaks at a single Rad52 repair centre. AB - DNA double-strand break repair (DSBR) is an essential process for preserving genomic integrity in all organisms. To investigate this process at the cellular level, we engineered a system of fluorescently marked DNA double-strand breaks (DSBs) in the yeast Saccharomyces cerevisiae to visualize in vivo DSBR in single cells. Using this system, we demonstrate for the first time that Rad52 DNA repair foci and DSBs colocalize. Time-lapse microscopy reveals that the relocalization of Rad52 protein into a focal assembly is a rapid and reversible process. In addition, analysis of DNA damage checkpoint-deficient cells provides direct evidence for coordination between DNA repair and subsequent release from checkpoint arrest. Finally, analyses of cells experiencing multiple DSBs demonstrate that Rad52 foci are centres of DNA repair capable of simultaneously recruiting more than one DSB. PMID- 12766778 TI - Differential regulation of E2F1 apoptotic target genes in response to DNA damage. AB - E2F1, a member of the E2F family of transcription factors, in addition to its established proliferative effect, has also been implicated in the induction of apoptosis through p53-dependent and p53-independent pathways. Several genes involved in the activation or execution of the apoptotic programme have recently been shown to be upregulated at the transcriptional level by E2F1 overexpression, including the genes encoding INK4a/ARF, Apaf-1, caspase 7 and p73 (refs 3-5). E2F1 is stabilized in response to DNA damage but it has not been established how this translates into the activation of specific subsets of E2F target genes. Here, we applied a chromatin immunoprecipitation approach to show that, in response to DNA damage, E2F1 is directed from cell cycle progression to apoptotic E2F target genes. We identify p73 as an important E2F1 apoptotic target gene in DNA damage response and we show that acetylation is required for E2F1 recruitment on the P1p73 promoter and for its transcriptional activation. PMID- 12766779 TI - Stroke services high priority, but neither coordinated nor timely. PMID- 12766780 TI - Asthma in New Zealand--time to get control. PMID- 12766782 TI - Asthma morbidity, control and treatment in New Zealand: results of the Patient Outcomes Management Survey (POMS), 2001. AB - AIMS: To determine the magnitude of morbidity from asthma within the New Zealand population, the degree of satisfaction of patients with their asthma control, and the level of asthma control achieved in relation to treatment. METHODS: Participants were randomly selected from 29 randomly chosen general practices throughout New Zealand. Information was collected from demographic and clinical questionnaires and from lung function tests. Criteria based on GINA guidelines were developed to define the level of asthma control for each participant, their opinion of their level of control, and to define which participants were under treated. RESULTS: A total of 445 patients (327 adults, age 16-68; 118 children, age 7-15) took part in the study. Ninety three per cent of adults had asthma that was sub-optimally controlled, 71% had asthma that was not well controlled, and 19% had asthma that was markedly out of control. For children, these figures were 90%, 42% and 4% respectively. These results were consistent regardless of asthma severity. In adults and children whose asthma was not well controlled, 49% and 71% respectively were under-treated. For those whose asthma was markedly out of control, 89% and 75% of adults and children respectively were under-treated. CONCLUSIONS: A significant proportion of patients have asthma that is not well controlled or that is markedly out of control, and the majority are under treated. PMID- 12766781 TI - Stroke rehabilitation services in New Zealand. AB - AIMS: To obtain an overall picture of the organisation of stroke rehabilitation services throughout New Zealand and to see if this is consistent with recommendations in evidence-based guidelines. METHODS: A questionnaire was sent to all hospitals in New Zealand. This included questions about access to organised stroke rehabilitation, guidelines for the management of common problems after stroke, and the use of audit. RESULTS: All 48 hospitals surveyed responded, with 37 providing inpatient stroke rehabilitation services. Only one hospital (serving 9% of the population) provided a dedicated, inpatient stroke rehabilitation facility. In the other 36 hospitals, stroke rehabilitation was performed in assessment, treatment and rehabilitation units (25 hospitals, 84%) or general medical wards (8 hospitals, 7%). Only 57% of the population had access to hospitals with a nominated lead clinician for stroke rehabilitation services. Thirty per cent were served by hospitals without a multidisciplinary therapy team expert in stroke care. Guidelines for the management of common problems following stroke were used in most hospitals. Only 8 hospitals (28%) had audited their stroke rehabilitation services. CONCLUSIONS: The organisation and type of rehabilitation services available for people with stroke are not consistent with best practice or accepted guidelines. The development of an organised approach to stroke rehabilitation services in New Zealand must be seen as a priority. PMID- 12766783 TI - The epidemiology of colorectal cancer: what can we learn from the New Zealand Cancer Registry? AB - AIM: Colorectal cancer (CRC) is a major cause of cancer death in New Zealand. Incidence and mortality rates are regularly reported but more detailed information concerning the epidemiology of the disease is not readily available. The aim of this study is to describe the epidemiology of colorectal cancer, with particular reference to gender; subsite distribution; stage at diagnosis; tumour grade; the contribution of recognisable aetiological factors; and the resection rate and type of surgery performed for rectal cancer. METHODS: All 2272 pathology reports of colorectal cancer submitted to the National Cancer Registry for the year 2000 were reviewed and restaged according to the classical Dukes system. Age, gender, stage, subsite distribution, differentiation, tumour type, associated pathology and type of rectal cancer specimen were recorded. RESULTS: The gender ratio for all cases was equal. One third of resected cancers were in the rectum, one third in the left colon and one third were proximal to the splenic flexure. Men had more rectal cancer (M:F 1.6:1) and women more colon cancer (0.9:1). Right-sided cancers were of significantly higher grade than left sided cancers. The proportion of right-sided cancers increased with advancing age in both men and women. Three per cent of patients had synchronous tumours and 0.2% of patients had phenotypic features of familial adenomatous polyposis (FAP). Fourteen per cent of patients were Dukes stage A at diagnosis, 43% stage B and 43% stage C. Rectal cancers were more likely to be stage A than colon cancers (odds ratio 1.6). Stage and grade of CRC were significantly associated. Ninety one per cent of cancers were resected; 29% of patients with rectal cancer had an abdominoperineal resection and 6% a local excision. CONCLUSIONS: The current stage distribution of CRC is in keeping with a predominantly unscreened population. The subsite distribution of cancer and its relationship to age and gender are similar to other high-risk countries; however, its relationship to grade of tumour has not previously been reported. An abdominoperineal resection rate of 29% reflects the mix of specialised colorectal and general surgical units treating rectal cancer in New Zealand and is in line with or better than recent overseas data that reflect national or regional practice. PMID- 12766784 TI - An audit of anaesthetic fresh-gas flow rates and volatile anaesthetic use in a teaching hospital. AB - AIM: The large number of anaesthetics administered means that the total cost to a hospital of inhalational anaesthetic agent such as isoflurane or sevoflurane can be considerable. The total anaesthetic gas flow is a major determinant of the use of these agents. Modern anaesthetic machines and monitoring facilitate reduced gas flows, which can significantly reduce wastage of these anaesthetic agents. The purpose of this study was to audit gas flow rates and volatile anaesthetic use. METHODS: We audited gas flows and choice of anaesthetic agent over two one month periods in one theatre at Christchurch Hospital. Data were collected directly from the anaesthetic machine using a computer. The second study period was clearly advised and followed widespread discussion of results from the first study period. RESULTS: Average fresh-gas flow was approximately 2 l/min (Month 1 = 2.0 l/min, Month 2 = 2.1 l/min). Use of the more expensive agent, sevoflurane, increased but gas flows with this agent decreased. CONCLUSIONS: Given the low flows used, the small difference between study periods was not surprising. The gas flows recorded represent responsible use of anaesthetic agents and are at least as good as flows achieved in previous studies that employed various methods to encourage their reduction. PMID- 12766785 TI - Controversies, new risk factors and the neuroendocrine approach to fibromyalgia: can therapeutic progress be made at last? PMID- 12766786 TI - Botulinum toxin as a preventive treatment for migraine and chronic tension headache. PMID- 12766788 TI - Death of pharmacy or safer dispensing? PMID- 12766787 TI - Primary hyperparathyroidism due to giant adenoma after treatment with radioactive iodine. PMID- 12766789 TI - Body composition in the elderly: the influence of nutritional factors and physical activity. AB - BACKGROUND: Controversy exists regarding the relative contribution of diet and exercise to body composition. Few studies have examined these associations in the elderly, where changes occur in the body fat to muscle ratio. OBJECTIVE: The primary objective of this paper is to determine whether energy intake or physical activity are associated with body composition. Secondly, to investigate whether specific macronutrients are associated with fat or lean tissue. DESIGN: Data (n= 1404) for this cross-sectional analysis were collected from a population-based sub-sample of elderly enrollees in the Cardiovascular Health Study (CHS). Dietary intake and physical activity were assessed by questionnaires. Body composition was measured by Dual Energy X-ray Absorptiometry (DEXA). Linear regression models were used to assess the associations of diet and activity with body composition. RESULTS: Total energy intake was not associated with any of the body composition measures. Higher dietary saturated fat was associated with higher percent body mass as fat and trunk fat in both sexes (p<0.01), and in men other dietary fats were associated with body fat. In women, distance walked was inversely associated with fat masses even after adjustment for pace of walking. In both sexes, faster pace of walking was associated with lower body and fat mass (p<0.01). Lean muscle mass was not associated with physical activity or dietary intakes. CONCLUSION: Physical activity and dietary fat intake in this the elderly population were more closely associated with body fat mass than was total energy intake. PMID- 12766790 TI - Nutritional status using the Mini Nutritional Assessment questionnaire and its relationship with bone quality in a population of institutionalized elderly women. AB - Malnutrition, a risk factor for osteoporotic fractures, is frequent in elderly people and, is underdiagnosed and undertreated. There are only few studies on the nutritional status of elderly people in Europe. The Mini Nutritional Assessment (MNA) is a non invasive and validated questionnaire to evaluate nutritional status in elderly people, classified in three groups: 1 degree score < 17: malnourished, 2 degrees score >17 and < 24: at risk of malnutrition, 3 degrees score >24: well-nourished, with a maximum of 30 points. Quantitative ultrasound of bone (QUS) is a method for assessing quality of bone which can be easily performed in nursing homes. Therefore, these two tests allowed to study the relationships between nutritional status and ultrasonic parameters of bone in 78 institutionalized women aged 86 +/- 6 years, living in 11 nursing homes around Lausanne (Switzerland). All were assessed by the MNA, had a measurement of the tricipital skin fold and of the grip strength. Functional status was evaluated by the scale "Activity of Daily Living" (ADL), and serum albumin level was measured when permitted. All had QUS of the calcaneus (with an Achilles, GE Lunar). The measured parameters are the Broadband Ultrasound Attenuation (BUA), attenuation of a band of ultrasonic frequencies through the medium, expressed in dB/MHz, and the Speed of Sound (SOS), speed of the ultrasounds through the medium, expressed in m/s. A third parameter, the stiffness index (SI), expressed as a percentage of the values obtained by the manufacturer in a young population and derived from BUA and SOS, was calculated automatically : SI = (0.67xBUA) + (0.28xSOS) - 420, expressed in percent compared to a young adult population (%YA). Fifteen percent of the women were undernourished and 58% were at risk of malnutrition. As expected, compared with the well-nourished minority, undernourished subjects had significant lower body mass index (BMI), tricipital skin fold (TSF), ADL score and albumin level (p < 0,01). The subjects "at risk of malnutrition" had significant lower BMI, ADL score (p < 0.01), tricipital skin fold and serum albumin (p < 0.05). Ultrasound parameters were low independently of the nutritional status. MNA score correlated significantly with tricipital skin fold (r = 0.508, p < 0.01), ADL (r = 0.538, p < 0.01) and albumin serum level (r = 0.409, p = 0.01). There was a trend for a correlation between the MNA and the ultrasound parameter BUA (r = 0.207, p = 0.07), whereas no correlation was found with SOS and SI. A multivariate analysis showed that tricipital skin fold and ADL explained 61% of the variance of the MNA. In conclusion, using simple and non invasive methods, this study showed that malnutrition and osteoporosis are frequent in institutionalized elderly persons in our country, and the ultrasound parameters are influenced by many others factors in addition to nutrition, especially at this age and in elderly residents of nursing homes. PMID- 12766792 TI - Homocysteine: a marker for cognitive performance? A longitudinal follow-up study. AB - The present prospective study investigated whether elevated total serum homocysteine concentration is a risk factor for cognitive decline. The outcomes were compared to the possible relation between cognition and vitamin B12 or folic acid. Cognitive performance of 144 normal aging individuals (aged 30-80 years) was tested at baseline and after six years of follow-up. Domains of cognitive function addressed were cognitive speed (Letter-Digit Coding test), attention and information processing (Stroop test) and verbal learning and memory (Word Learning Test Total; Delayed Recall). Serum concentrations of homocysteine, folic acid and vitamin B12 were determined. Serum concentrations of homocysteine correlated negatively with cognitive performance on the Word Learning tests at baseline, independent of age, sex, education level or folic acid concentration. Homocysteine concentration at baseline correlated negatively with cognitive performance on the Stroop and Word Learning tests during the whole six-year follow-up period. The folic acid concentration correlated to the Delayed Recall test at baseline only and no correlations were observed for vitamin B12. Thus, while a relation between vitamin B12 or folic acid and cognition was almost absent, elevated homocysteine concentrations were associated with prolonged lower cognitive performance in this normal aging population. PMID- 12766793 TI - Plasma total homocysteine levels, dietary vitamin B6 and folate intake in AD and healthy aging. AB - PURPOSE: To study the association between Alzheimer s disease (AD) and plasma total homocysteine (tHcy), dietary folate and vitamin B6. METHODS: 64 AD patients were matched by gender, age, and smoking status to 64 healthy controls. tHcy was determined using an automated immunoassay. Dietary patterns for three age periods (20-39, 40-59, and 60 + yrs) were assessed using a questionnaire adapted from the Block Health Habits and History Questionnaire. Respondents (cases by proxy) reported food frequencies, which were translated into estimated daily nutrient intakes. APOE genotype, cognitive performance (CDR, MMSE), blood lipids, and albumin were obtained for patients and controls. RESULTS: tHcy did not differ significantly between controls (11.5 +/- 3.7 mmol/L) and AD patients (12.3 +/- 4.3 mmol/L)(p=0.25). tHcy levels were not related in AD patients or controls to education, CDR, MMSE, blood lipids, albumin or ApoE genotype (p>0.15). There was a negative correlation between plasma tHcy and triglyceride levels in AD patients (p=0.023), but not in controls. AD patients consumed significantly less dietary vitamin B6 (p=0.05) and folate (p=0.001) after age 60 than controls. CONCLUSIONS: Although plasma tHcy levels were higher in cases than controls, this difference was not significant. tHcy levels were not related to cognitive status. Plasma tHcy was inversely correlated with triglyceride levels in AD patients but not in controls. PMID- 12766794 TI - Influence of antidepressant therapies on weight and appetite in the elderly. AB - OBJECTIVE: Determine the nutritional impact of antidepressant drugs in elderly. METHODS: The study included 139 independently mobile out-patients managed by the Poitiers geriatric day hospital for problems of loss of home support, rehabilitation and medical or psychological care, with a stable treatment for 3 months at the time of inclusion and over the 3 months of the study. The study addressed the time course of nutritional parameters from baseline to 3 months post-inclusion. The physical examination included monthly weighing of the patients, 3-monthly evaluation of nutritional status using the Mini Nutritional Assessment (MNA) instrument and serum albumin. RESULTS: 52 men (76.3 +/- 6.7 years [62-87]) and 87 women (81.0 +/- 7.2 years [65-100]) were included. Seventy nine patients presented with dementia. Seventy-four patients were receiving an antidepressant, of which 54 serotonin reuptake inhibitors. The patients free from dementia and not receiving antidepressants had nutritional indices that did not vary over the study period. The dementia-free patients receiving antidepressants gained weight (1.44 kg) - 1.87 kg on serotonin reuptake inhibitors - showed an improvement in MNA of 0.76/30 and showed a significant improvement in serum albumin of 1.78 g/L. The demented patients not receiving antidepressants lost weight (-1.01 kg), MNA score fell and serum albumin significantly decreased. On antidepressants, the demented patients significantly gained weight (0.73 kg) while no deterioration in the other parameters reflecting undernutrition was observed. Conclusions. In the middle term, in elderly subjects, antidepressants do not induce undernutrition or weight loss. Irrespective of antidepressant type, those agents seem to prevent weight loss in elderly subjects presenting with dementia. PMID- 12766795 TI - Longitudinal study of fasting serum glucose concentrations in healthy elderly. The New Mexico Aging Process Study. AB - BACKGROUND: The American Diabetes Association s Expert Committee on the Diagnosis and Classification of Diabetes Mellitus has made the recommendation that all individuals over the age of 45 years should be screened for diabetes every 3 years. OBJECTIVE: This study was designed to determine the necessity for screening healthy elderly (> 65 years) this frequently using fasting serum glucose (FSG) determinations. DESIGN: This is a longitudinal study of initially healthy, upper middle class, community-based volunteers, mostly age 65 years and older at entry into the study. Participants were followed longitudinally with annual FSG concentrations and body mass indices (BMI) for periods up to 18 years (mean 12.4 years). RESULTS: Only 4 of 299 individuals with entry FSG < 126 mg/dl (mean + S.D. age at entry 71.6 + 4.8 years) and 6 or more annual visits have subsequently met the Expert Committee criteria for the diagnosis of diabetes (two consecutive FSGs > 126 mg/dl unless under treatment). When one examines the slopes of FSGs plotted over time (years) for each individual, more participants had a negative slope (220) than positive slope (79), i.e., their FSGs tended to decrease with age. None of the 68 individuals entered age > 75 years subsequently developed diabetes or a significantly positive slope. CONCLUSIONS: It does not appear necessary to screen non-obese elders (excluding minorities) age >65 years with a FSG < 100 mg/dl, or those age >75 years every 3 years as recommended. PMID- 12766796 TI - Evaluation of the nutritional status of patients over 60-year admitted in a hematology department using the mininutritional assessment (MNA). A single centre study of 120 cases. AB - BACKGROUND: The nutritional status of older persons suffering from various medical or surgical conditions has been well reported. However, studies focusing on hematology patients are very rare. OBJECTIVE: Our aim was to obtain a photographic picture of the nutritional profile of a sample population treated for miscellaneous blood disorders, to compare our results with those obtained in other types of diseases and to isolate risk-factors for undernutrition. DESIGN: One hundred-twenty free living hematology patients aged over 60-year were prospectively and randomly tested the day of their admission to either our out- or inpatient clinic (department of clinical hematology from a French general hospital) using the mininutritional assessment (MNA). We compared our data with those from the literature and the role of various risk-factors was evaluated using the chi-square or the Fisher exact test. RESULTS: The sex ratio was 1 and the median age 74 (range: 60-97). The majority of this population suffered from malignant disorders (101 cases, 84%) and fifty-three of them received chemotherapy. Eighty individuals (66%) were tested at the outpatient unit. The mean MNA score was 22.8 (range: 7.5-30). Sixteen patients (13%) were categorized as in poor nutritional status (MNA< 17). In this subgroup, more than 3 drugs intake (p < 0.01) and recent weight loss (p = 0.015) were the most important MNA parameters predicting malnutrition (age, sex, disease type or duration being no significant risk-factors). Our MNA results compared favorably with those obtained in other medical or surgical specialties. CONCLUSION: Undernutrition does not appear more prevalent in the elderly hematology population in comparison with patients suffering from other diseases. However, because nutritional status may influence the outcome, especially in case of blood neoplasms, MNA (or other more sophisticated biological tools) should be included in the evaluation of this population. PMID- 12766798 TI - A high-resolution genetic and physical map of a mouse coat abnormality locus (Uncv). AB - More than a hundred of loci (genes) affect the development of mouse and human hair. A locus of Uncv (uncovered) has been confirmed to be involved in hairlessness for homozygote and sparse hair for heterozygote. Except hairlessness(or called uncovered coat), the homozygote was also accompanied by growth retard and puberty delay. Identification of the mutation in the gene will be important for understanding the related diseases in human. Although the uncovered locus (Uncv) has been mapped to the mouse distal chromosome 11(Chr11), the high-resolution genetic map and physical map of the locus has not been created. In this study, 2074 F2 mouse populations from backcross [BALB/c (Uncv/Uncv) x C3H (+/+)]x BALB/c (Uncv/Uncv) and [BALB/c (Uncv/Uncv) x C57BL/6 (+/+)] x BALB/c(Uncv/ Uncv) were genotyped using 16 polymorphic markers with an approximately 20 cM interval on mouse distal Chr11. By genetic linkage analysis, Uncv locus was mapped to an approximately 1.4 cM interval between markers D11Mit337 and D11Mit338 with the following order: proximal D11Mit338-D11Mit203 (Uncv)-D11Mit103 -D11Mit337 distal on mouse Chr11. And then, a contig of 35 BACs representing the Uncv-containing region was constructed. The contig covered 800 1000 kb region flanked by 189K10-SP6 and D11Mit103. Together, we have constructed the high-resolution genetic map and detailed physical map of the Uncv region. This will facilitate the identification of the Uncv loci. PMID- 12766799 TI - Functional expression of guinea pig growth hormone receptor and its mutants in mammalian cells. AB - The cDNA of guinea pig (Cavia porcellus) growth hormone receptor (gpGHR) was cloned using RT-PCR in our laboratory. By sequence alignment, substitutions of amino acids conserved in other mammalian GHRs were found. For example, histidine 168 and tyrosine-332 equivalent to positions 170 and 333 in other mammalian GHRs, which were considered to be necessary for the dimerization of GHR and the specific GH-stimulated functions respectively, were replaced by tyrosine and serine in gpGHR. Here, we report the functional expression of gpGHR and its mutants, gpGHRY168H and gpGHRS332Y, in COS-7 cells and/or Chinese hamster ovary (CHO) cells. It was shown that the COS-7 cells transfected with pcDNA3-gpGHR possessed high affinity to bovine GH [K(a) = 1.3 x10(9) (mol/L)(-1)] and a protein band with molecular weight around 92 kD was detected by anti-mouse GHR monoclonal antibody (mAb263). When CHO cells were transfected with the expression vectors, pcDNA3-gpGHR, pcDNA3-gpGHRY168H and pcDNA3-gpGHRS332Y, the gpGHR and its mutants were expressed and the ligand binding, phosphorylation of JAK2, protein synthesis, and lipogenesis were studied. It was found that the mutation of serine to tyrosine at position 332 greatly increased the GH-stimulated protein synthesis and the phosphorylation of JAK2, while the mutation of tyrosine to histidine at position 168 increased the protein synthesis and decreased the phosphorylation of JAK2 only weakly. However, both mutations decreased the GH-stimulated lipogenesis. Thus, our study provides the experimental evidence that gpGHR may mediate the metabolic actions of GH and the substitutions of some conserved amino acids in gpGHR result in the changes of post-binding signaling. PMID- 12766800 TI - Identification of important amino acid residues for human IL-18 function by mutant construction. AB - To study the structure-function relationship of IL-18, two IL-18 mutants, N- and C-terminal mutant (Delta NC) and IL-1 signature-like sequence mutant S(154)A/Y(156)F/E(157)P/C(163)T (S), were constructed by PCR. The wild type and mutant recombinant human interleukin-18 (rhIL-18) were expressed in E.coli, purified by Sephadex G-75 chromatography and renatured by stepwise dilution. The purity of the recombinant proteins was over 95%. The activities of wild type and mutant rhIL-18s were defined as the ability to induce interferon-gamma (IFN gamma) production and NF-kappa B activation from human peripheral blood mononuclear cells (PBMC). Our results showed that the two mutants induced significantly less amount of IFN-gamma from PBMC (13%, 48% of wild type rhIL-18 for Delta NC, S, respectively), and the activation of NF-kappa B also lower than wild type rhIL-18(69.7%, 89.8% of wild type rhIL-18 respectively), indicating that the deleted or mutated amino acids might be important for IL-18 function. PMID- 12766801 TI - Purification and characterization of Alcaligenes faecalis penicillin G acylase expressed in Bacillus subtilis. AB - The Alcaligenes faecalis PGA gene encoding heterodimeric penicillin G acylase (PGA) was cloned and successfully expressed in Escherichia coli and Bacillus subtilis respectively. In contrast to E.coli hosts where the enzymes were retained in the periplasm, B. subtilis cell secreted the recombinant enzyme into the medium. Contrary to limited expression yield of E. coli (pETAPGA), PGA extracellularly expressed by B. subtilis (pBAPGA) and B. subtilis (pMAPGA) reached the highest yield of 653 u/L. This yield increased 109-fold higher than the native expression of A. faecalis CICC AS1.767. The enzyme was fractionated with (NH(4))(2)SO(4) and purified by DEAE-Sepharose CL-6B with a yield of 81%. The purified enzyme had a specific activity of 1.469 u/mg. Furthermore, some enzyme characteristics, such as the pH and temperature optimum, the stability against organic solvent and the ratio of cepholexin synthesis to hydrolysis were determined. By overexpressing A. faecalis PGA in B. subtilis and purifying secreted enzyme from culture medium one could readily obtain a large amount of an alternative source of PGA. PMID- 12766802 TI - Induced expression of the gene for NADP-malic enzyme in leaves of Aloe vera L. under salt stress. AB - A cDNA fragment for NADP-malic enzyme, catalyzing the reversible oxidative decarboxylation of L-malate to produce CO(2), pyruvate and NADPH, was isolated from the leaves of a 2-month-old Aloe vera L., The level of expression of NADP-ME mRNA and accumulation of NADP-ME (AvME) protein under salt stress conditions were compared between a tolerant aloe, Aloe vera L. and a sensitive aloe, Aloe saponarea Haw. The results suggested that both the expression of the gene and the accumulation of the protein were induced in the two kinds of aloe, and the strength was related to the degree of salt tolerance. Northern blot analysis revealed that the gene for NADP-malic enzyme in Aloe vera L.( AvME) was induced by high salt, dehydration, and exogenous abscisic acid (ABA), but not by cold treatment. To further confirm whether the synthesis of AvME protein was induced with hours of treatment, Western blot analysis of the samples was conducted. The results indicated that the induction of AvME protein expression was obvious after 48 h at high salt and the level was increased with the hours of treatment. PMID- 12766803 TI - Effects of CD/5-FC suicide gene therapy system on human malignant glioma cells in vitro. AB - The purpose of this paper is to investigate the antitumor effects of cytosine deaminase/5-fluorocytosine (CD/5-FC) suicide gene therapy system on human malignant glioma cells in vitro. The pCMVCD plasmid was constructed through the CD gene insertion in the multicloning site of eukaryotic expression vector pcDNA3.0, and confirmed by restriction endonuclease digestion/gene sequencing. The construct was subsequently transfected into the U251 human malignant glioma cells by using LipofectAMINE2000-mediated method. Resistant clones (named U251/CD cells) were isolated by screening with G418 presence. U251/CD cells were incubated with 5-FC in different concentrations to determine viability ratios (or cytotoxicity assay), measured by the 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) assay. The concentrations of 5-fluorouracil (5 FU) in the media were measured by high-performance liquid chromatography (HPLC) detector. Our results suggested that the untreated U251 cells were insensitive to 5-FC, with the IC(50) about 6500 micromol/L. After transfection, the IC(50) was dramatically reduced to about 10 micromol/L. Therefore, gene transfection made G418-resistant clones (U251/CD cells) be highly sensitive to 5-FC. HPLC analysis showed that 5-FU was detected in U251/CD cell medium. Study on U251 cells genetically modified by CD gene in vitro will play an essential role in glioma gene therapy in vivo. In conclusion, our results indicated that the CD/5-FC system was feasible to treat glioma. PMID- 12766804 TI - Conditional gene activation in cultured hepatocytes using a ligand-dependent chimeric Cre recombinase. AB - By combining liver-specific promoter and a chimeric Cre recombinase, conditional gene activation could be finely achieved in hepatocytes at selected time points. To this end, the expression vector of Cre-ERt under the control of the mouse albumin gene promoter/enhancer, alb-Cre-ERt, was constructed, and transfected into engineering BRL (Rat hepatocytes) and BRK (Rat kidney) reporter cells which carries a chromosomally integrated 'floxed' beta geo gene, which is inserted between the promoter and the human alkaline phosphatase( hAP) reporter gene, thereby preventing hAP reporter gene transcription, respectively. After treatment with 1 micromol/L 4-hydroxytamoxifen(4-OHT), a proportion of hAP staining positive cells were detected by hAP staining. It was further confirmed that 'floxed' beta geo cassette was removed by Cre excision by using PCR analysis of cellular DNA. No background recombinase activity could be detected in the absence of 4-OHT. Moreover, no hAP-positive cells could be detected in BHK cells untreated or treated with 4-OHT. These data suggested that alb-Cre-ERt expression vector was constructed successfully, and 4-OHT could induce Cre-mediated recombination only in hepatocytes expressing Cre-ERt, thereby activating a stably integrated hAP reporter gene. This provides a further foundation for producing transgenic mice expressing such an 4-OHT inducible Cre recombinase specifically in mouse liver. PMID- 12766805 TI - [Optimization of T7-based RNA amplification system for cDNA microarray]. AB - cDNA microarrays are powerful parallel tools for gene expression profiling analysis, which help us to understand the molecular mechanism of diseases and to identify potential targets for therapeutic intervention. However, their broader application are hampered by the large amount of RNA required: up to 200 microg of total RNA or 5 microg of mRNA for one chip, making analysis of small samples difficult. In this work, combined with a template switching effect, the T7 RNA linear amplification procedure was optimized, providing multiple copies of anti sense RNA with reduced sample inputs: no more than 3 microg total RNA for one chip. Using the same RNA sample in all cases, the anti-sense RNA labeling method was compared with standard total RNA and mRNA methods by two sets of self comparison experiments. Furthermore, the three methods in profiling analysis were compared with the same pair RNA samples. All the results indicated that the fidelity, reproducibility, and reliability showed no significant difference with conventional total RNA or mRNA microarrays. PMID- 12766806 TI - [Reconstruction of the three-dimensional structure of senile plaques in Alzheimer disease]. AB - Senile plaques are one of the major neuropathological features of Alzheimer's disease (AD). Aggregation of Amyloid beta-protein(A beta) in senile plaques is related to the pathogenesis of AD, so it is very important for the studies on the prevention of A beta deposition and on trying to disaggregate the senile plaques, aimed at treatment and prevention of the AD. In this report, the three-dimension structures of senile plaques are reconstructed by AVS software by using immunohistochemical method and MATLAB software. The diffuse plaques and classical plaques in hippocampus all have complicated porous structures; no core structures were present in diffuse plaques, and the deposition density of A beta in this kind of plaque diminishes from interior region to the microenvironment. The deposition density of A beta was highest in the central region of classical plaques, while was lowest in the region around central, and was intermediate in the extreme region of this kind of plaques. The method of reconstruction of three dimension structure of senile plaques helps studying on dynamics of senile plaques, and it is also important for the studies on the pathogenesis of AD. PMID- 12766807 TI - [Secretory expression of an antifungal protein in the mold Trichoderma viride]. AB - AFP (antifungal protein) secreted by the filamentous fungi Aspergillus giganteus MDH18894 is a strong basic protein (pI>10.65). Its mRNA has an 430 bp open reading frame encoding the 94 amino acids' precursor of AFP. The precursor of AFP is processed to form the mature AFP of 51 amino acids by a putative two-step processing. A 285 bp AFP-like gene (i.e. AFP cDNA) lacking introns was reported to exist in the genome of Trichoderma viride, another species of filamentous fungi, but the gene was silent. In order to investigate whether the genomic AFP gene from A. giganteus could be expressed in T. viride, an expression plasmid with the open reading frame of AFP gene fused between the promoter and terminator of trpC gene was constructed and was transformed into T. viride. SDS-PAGE and Western blot analysis showed that AFP was expressed and secreted into the culture supernatant of the transformant of T. viride. Our study provides a basis for further secretory expression of heterologous eukaryotic genes of important value in the system of T. viride. PMID- 12766808 TI - [Analysis of protein phosphorylation by combination of IMAC, phosphatase with biological mass spectrometry]. AB - Protein phosphorylation is the most important reversible post-translational modification in cells. Analysis of phosphorylated proteins and identification of their phosphorylation sites is helpful for understanding their biological functions. MALDI-TOF-MS and ESI-Q-TOF-MS play important roles in protein phosphorylation analysis. In this work, immobilized metal affinity chromatography (IMAC) was used to selectively enrich phosphopeptides from protein digest mixtures, and treatment of phosphopeptides with alkaline phosphatase was used to confirm the phosphorylation. Finally, the phosphorylation sites were determined by tandem mass spectrometry analysis and database searching. PMID- 12766809 TI - [The preparation of an artificial red blood cell substitute by W/O/W double emulsion methods]. AB - A method of preparing methoxypoly (ethylene glycol)-poly-DL-lactide (PELA) microcapsule containing bovine hemoglobin(BHb) by using W/O/W double emulsion solvent diffusion process for use as artificial red cells was developed. The results suggested that preparation conditions such as solidification method, stirring rate, type of organic solvent had significant influence on the encapsulation efficiency and the activity of bovine hemoglobin. When ethyl acetate was used as the organic solvent and double emulsion-solvent diffusion process was used, there was no significant influence on the activity of hemoglobin if stirring rate was lower than 9000 r/min. High stirring rate, 12,000 r/min, decreased the P(50) and Hill coefficient of the hemoglobin encapsulated. Increasing the volume of solidification solution had an effect of improving the activity of hemoglobin to carry oxygen. PMID- 12766810 TI - [Purification and characterization of alginate lyase from marine bacterium Vibrio sp. QY101]. AB - Extracellular alginate lyase secreted by Vibrio sp. QY101, which was isolated from brown algae, was purified to homogeneity by a combination of ammonium sulfate precipitation, DEAE-Sepharose Fast Flow anion-exchange chromatography and Superdex 75 gel filtration chromatography. Its molecular mass was 39 kD as determined by SDS-PAGE analysis. The enzyme had an optimal temperature of 30 degrees for its activity, and was most active at pH 7.5. The thermal and pH stability were 0-30 degrees, and pH 6.5-8.5, respectively. The enzyme activity was stimulated by 0.5 mol/L NaCl, 1.0 mmol/L Ca(2+) or 5.0 mmol/L (Mn(2+), and inhibited by 5.0 mmol/L Ni(2+), 1.0 mmol/L Fe2+) or 1.0 mmol/L EDTA. Preliminary analysis on substrate specificity showed that this alginate lyase had activity on both poly-alpha 1,4-L-guluronate and poly-beta1,4-D-mannuronate substrates. PMID- 12766811 TI - [Screening of proteins interacting with tTG in HeLa cells]. AB - Tissue transglutaminase(tTG) belongs to a class of transglutaminase family which is up-regulated in almost all cells apoptosis and is thought to be closely related to cell apoptosis. To investigate the mechanism of tTG in cell apoptosis, yeast two hybrid system was used to screen HeLa cDNA library. One of the 17 positive clones we have obtained encoded the glutamine-rich carboxyl terminus of TIAR, and this interaction between tTG and TIAR, which was finely regulated by Ca(2+), was proved in vitro by GST pull-down. These findings suggest that tTG might affect the function of TIAR by a calcium-dependent posttranslational modification and the interaction might possibly be involved in the regulation of cell apoptosis. PMID- 12766812 TI - [Expression and purification of the N-domain of human canstatin and its bioactivity]. AB - Total RNA was extracted from placenta umbilical tissue. The canstatin cDNA was amplified from total RNA by net-RT-PCR technique and cloned into pSP72, and the resulted plasmid pSP72C was used as template to amplify its N-domain. The amplified 1-89 aa N-domain was then cloned into pET-3c. The resulted plasmid pET CN was transformed into E. coli BL21(DE3). The N-domain was efficiently expressed after IPTG induction as a 10 kD band on SDS-PAGE. The expressed product accounted for approximately 35.3% of the total bacterial proteins, as estimated by densitometry and existed mainly as inclusion body. The inclusion bodies were washed, lysed and the reactivated proteins were purified by the Sephadex G-100 gel filtration to a purity of 92.6%. CAM assay showed that N-domain effectively inhibited the angiogenesis of chicken embryo microcapillary vessel. PMID- 12766817 TI - [Evulsion of the optic nerve following blunt bulbar trauma. Case report and review of the literature]. AB - BACKGROUND: Evulsion of the optic nerve is a rare form of traumatic optic neuropathy. It is a rupture of the optic nerve at the disc without damage of its sheaths occurring in association with a blunt skull trauma or a blunt bulbar trauma. METHOD: We report on a 13-year old girl with complete evulsion of the optic nerve after blunt bulbar trauma and give a literature review. RESULTS: While swinging on a liana the patient had crashed into a tree. The initial examination 3 hours after the accident revealed at the right eye an amaurosis without any afferent reaction of the pupil to light. Ophthalmoscopy displayed a vitreous haemorrhage at the optic disc, peripapillar white retina and interruption of the blood flow of all visible retinal vessels. Fluorescein angiogram showed no retinal perfusion and a detatched retina above the disc. Echographic examination revealed a vitreous haemorrhage at the disc shaped like a mushroom. From the eighth day, examination of the retina became impossible due to disseminating vitreous haemorrhage. After 8 months vitreous haemorrhage had disappeared revealing a white fibrosis at the area of the optic disc. Since first histological description of an optic nerve evulsion by His in 1856, 42 patients with complete and 23 with partial evulsion of the optic nerve were reported in the literature. In our review different mechanisms of injury are discussed. CONCLUSION: Evulsion of the optic nerve is rarely reported. It is possible that it may occur more frequently, because it cannot be seen in blunt or also penetrating bulbar traumas with severe intraocular cloudiness. PMID- 12766818 TI - [First steps with the Zeiss IOLMaster: A comparison between acoustic contact biometry and non-contact optical biometry]. AB - BACKGROUND: After having received an IOL-Master (Zeiss, Jena), we compared our actual standard method with this new device. PATIENTS, MATERIALS AND METHODS: From March to June 2002, we examined 175 consecutive cataractous eyes with the IOL-Master as well as with the acoustic biometry (Tomey AL-1000) combined to the Javal-Keratometer (Haag-Streit, Bern). The results were compared and analysed statistically. In all eyes, the intraocular lens to be implanted was chosen by means of the SRK/T formula, based on the measurements conducted with our standard method. The achieved postoperative refraction of 153 eyes obtained, at least six weeks after surgery, by the treating ophthalmologists, was communicated to us. RESULTS: Comparison of eye lengths as well as of the keratometric measurements showed good correspondence between the obtained measurements by both methods, acoustic biometry yielding significantly (p < 0.001) shorter axial lengths than the IOL-Master, and the Javal yielding significantly (p < 0.001) higher mean corneal refraction power than the IOL-Master. For both measurements, regression lines showed good coherence of the results over the refraction sample of our patients. Surprisingly, the accuracy of the refraction obtained postoperatively compared to the preoperative aim was better with our standard method compared to the IOL-Master. CONCLUSIONS: The predicted systemic differences in measurement results could be verified. No improvement in accuracy of our postoperative refraction prediction was achieved so far. The current advantage of the IOL Master in our clinic is the substantial gain in time, as well as the fact that performance of the measurements may be delegated. PMID- 12766819 TI - [Diagnostic and therapeutic problems in chronic progressive external ophthalmoplegia (CPEO)]. AB - BACKGROUND: Strictly speaking, CPEO is defined as paralysis of the external ocular muscles combined with ptosis. In Kearns-Sayre syndrome additional neurological or muscular deficits can be observed, sometimes even decades after the onset of ocular symptoms. METHOD: Three patients with classical CPEO and one patient with Kearns-Sayre syndrome where included in the present study. The clinical examinations included electromyography, various serological parameters and histological biopsies. RESULTS: The onset of ocular symptoms with divergent strabism and ptosis could be dated back to the age of 7, 27, 38 and 44 years starting on one eye and progressing slowly over years to both eyes. In the Kearns Sayre syndrome patient a general decrease in cardio-respiratory fitness and occasional collapses were noted. In three patients ptosis and strabism surgery led to the relief of the ocular symptoms. CONCLUSIONS: In CPEO respectively Kearns-Sayre syndrome the onset of the disease is variable. The final correct diagnosis is often delayed due to the mild beginning of the symptoms and the slow progression of the paralyses. Early EMG and muscle biopsy examination may facilitate the diagnosis. PMID- 12766820 TI - [Refractive benefit and incremental costs of LASIK: results of a cost benefit study in two universities' LASIK departments]. AB - BACKGROUND: No data is available to date on the cost effectiveness of laser in situ keratomileusis (LASIK) with regard to the German-speaking health care system. Hence we compared the clinical outcome and cost data of two German LASIK centres in respect of procedure cost effectiveness from the patient perspective. MATERIAL AND METHODS: A retrospective cohort study was implemented at the University Hospitals of Mainz and Mannheim. Data on refractive outcome were obtained from the LASIK centres' patient documentation, cost data on the surgery as well as on eyeglasses etc. before and after the surgery were obtained from patient interviews. The primary endpoint of this investigation was the individual incremental cost effectiveness ratio linking direct cost with the LASIK associated refractive gain. RESULTS: Clinical and economical data of 178 patients (Mainz: 45, Mannheim 133) was evaluated. After biplanar simultaneous LASIK 71% (Mainz) and 73% (Mannheim) reported, that they did not need eyeglasses etc. any longer after surgery; 31% (Mainz) versus 13% (Mannheim) had to undergo LASIK re treatment. Median direct cost for the surgery was reported to be 3000 euro; (Mainz) versus 3910 euro; (Mannheim); the corresponding LASIK associated refractive gain was +5.6 D (median Mainz) versus +4.2 D (median Mannheim), resulting in median incremental cost effectiveness ratios of 667 euro; (Mainz) versus 831 euro; (Mannheim) per gained refractive unit. This investment has to be related to median annual cost of 250 euro; for eyeglasses etc. before surgery and 0 euro; afterwards. CONCLUSION: Depending on the performing surgical centre, German LASIK patients have to face a median investment of 667-831 euro; per gained refractive benefit unit. This incremental cost benefit ratio can be regarded as a surrogate parameter for comparison with alternative surgical or conservative treatment strategies in myopia. PMID- 12766821 TI - [Cataract and keratoplasty--simultaneous or sequential surgery?]. AB - BACKGROUND AND PURPOSE: Since the introduction of the triple procedure (simultaneous penetrating keratoplasty [PK], extracapsular cataract extraction [CE] and implantation of a posterior chamber intraocular lens [PCIOL]) in the mid seventies, there is an ongoing discussion among corneal surgeons concerning the best approach for combined corneal disease and cataract. METHODS: Besides the classical triple procedure (1), two alternative microsurgical approaches are feasible: (2) CE + PCIOL prior to PK and (3) CE + PCIOL after PK. For the refractive results after TRIPLE some intraoperative details are crucial: Trephination of recipient and donor from the epithelial side without major oversize (Guided Trephine System or Nonmechanical Excimer Laser Trephination) should preserve the preoperative corneal curvature. Graft and the PCIOL placed in the bag after continuous curvilinear capsulorhexis should be centered along the optical axis. If possible, performing the capsulorhexis under controlled intraocular pressure conditions prior to trephination may help to minimise the risk of capsular ruptures. RESULTS: The major advantage of the TRIPLE is the faster visual rehabilitation and less efforts for the mostly elderly patients. However, two intraocular interventions with approach (2) and (3) bear an increased risk of infection and suprachoroidal haemorrhage. Approach (2) requires a cornea that is still transparent enough to perform cataract surgery, and the risk of intraocular pressure rise after PK seems to be increased. Approach (3) has the potential of a simultaneous reduction of astigmatism during CE (appropriate location of the incision, simultaneous refractive keratotomies or implantation of a toric PCIOL). Disadvantages may include the loss of graft endothelial cells and the theoretically increased risk of immunological allograft reactions. After TRIPLE, major deviations from target refraction have been reported. However, individual multiple regression analysis may help to minimise this problem with appropriate methods of trephination. Since suture removal after PK may result in major individual changes of the corneal curvature, IOL power calculation for approach (3) requires all sutures to be removed at the time of CE. However, even after complete suture removal the abnormal proportions between anterior and posterior curvatures and/or the irregular topographies after PK may be responsible for marked IOL power miscalculations in the individual case. CONCLUSIONS: The postulated better refractive outcome and better uncorrected visual acuity after the sequential approach is opposed by a markedly delayed visual rehabilitation. For this reason, we consider the TRIPLE procedure including CE via open sky in general anesthesia as the method of choice for combined lens and corneal opacities. Because of the often rapidly progressive nuclear cataracts after PK, we recommend the simultaneous approach in elderly patients with Fuchs' dystrophy even with incipient lens opacities. PMID- 12766822 TI - [Therapy of normal tension glaucoma: effect of brinzolamide on ocular haemodynamics]. AB - BACKGROUND: Altered ocular perfusion plays a role in the pathophysiology of normal tension glaucoma. Dorzolamide, a locally applied inhibitor of carbonic anhydrase, is thought to increase ocular blood flow. Less data are available regarding the influence exercised on ocular perfusion by brinzolamide, another and different, locally administered, inhibitor of carbonic anhydrase. PATIENTS AND METHODS: n = 15 eyes of 8 normal tension glaucoma patients were subjected to colour Doppler imaging and Langham-OBF (LOBF) before and during a therapy for 3 - 5 weeks with brinzolamide. RESULTS: Brinzolamide reduces intraocular pressure from 15.8 +/- 0.9 to 12.6 +/- 0.9 mm Hg (n = 15; P < 0.05). Systolic as well as diastolic blood flow velocities, resistive (RI) and pulsatility index (PI), measured by CDI, remained unchanged in the presence of brinzolamide. LOBF values are also not influenced by brinzolamide (1014 + 115 before vs. 1113 +/- 178 microl under therapy; n = 15; n. s.). DISCUSSION: Brinzolamide does not exercise any impact on ocular haemodynamics. This is different from the properties of dorzolamide that had been reported previously. PMID- 12766823 TI - [Anatomical differences in optic nerve, chiasma and tractus opticus in human albinism as demonstrated by standardised clinical and MRI evaluation]. AB - BACKGROUND: There has been long-standing clinical and electrophysiological evidence that in patients with albinism the visual pathways cross atypically: most fibres from one eye cross to the contralateral visual cortex. PURPOSE OF THE STUDY: to determine whether the size and configuration of the optic chiasm in human albinos is different from normally pigmented controls. PATIENTS AND METHODS: 17 patients (11 female, mean age 35.8 years) with oculocutaneous albinism underwent a standardised graded morphological and functional evaluation. Magnetic resonance images were reformatted to the region of the optic chiasm and analysed using observer-independent morphometry. In addition, fMRI of the visual cortex was performed during VEP analysis (1.5 Tesla Siemens Vision). Morphological and fMRT results were compared to an age-correlated group of n = 16 normally pigmented healthy volunteers with normal visual acuity and stereopsis. RESULTS: 65 % of the patients (n = 10) showed signs of dysplasia of the optic nerve head. Statistical morphometry showed distinct differences in chiasmal morphology between albinos and normally pigmented probands (smaller optic nerves, different angles of optical entry into the chiasm and of the beginning of the Tractus optici leaving the chiasm, overall chiasmal width and height). CONCLUSIONS: Optic nerve head anomalies are frequent in albinism and influence visual outcome. Size and configuration of the optic chiasm in human albinos is distinctly different from normally pigmented control persons and reflects the atypical crossing of optic fibres. PMID- 12766824 TI - [Development of ocular motility following modified 3-wall decompression of the orbita in endocrine orbitopathy for functional and rehabilitative indication]. AB - INTRODUCTION: Permanent visual damage due to an increase in volume of the orbital contents may be the result of the failure of conservative therapeutic concepts in the treatment of endocrine orbitopathy. Considerable progress has been achieved in developing successful orbital decompression techniques with regard to functional and cosmetic outcome. Decompression techniques with resection of the bony orbital walls are adequate tools in restoring visual acuity and reducing exophthalmus. A considerable degree of deterioration of motility disorders has been described in the literature depending on the techniques being used. PURPOSE: The purpose of this study was to investigate whether a modified technique of 3 wall orbital decompression with preservation of a medial part of the periorbital tissue to support the medial rectus muscle, is able to reduce the postoperative risk of diplopia. MATERIAL AND METHODS: A modified technique of orbital 3-wall decompression with resection of the medial orbital wall, the medial orbital floor and the floor of the frontal sinus has been used in patients with compressive optic neuropathy (n = 20) and for cosmetic reasons (n = 7) in cases of uni- or bilateral proptosis. Analysis of the results was performed concerning visual outcome, exophthalmus reduction and development of horizontal and vertical motility changes. RESULTS: In all cases of optic neuropathy improvement of visual function at an average of 4.63 +/- 4.5 lines could be achieved. Exophthalmus reduction was 3.2 +/- 2.4 mm in the functional group and 3.9 +/- 1.7 mm in the rehabilitative group. In this group motility of the medial rectus muscle remained unaffected except in one eye. In the functional group motility deterioration was observed in 62 %. CONCLUSION: The modified 3-wall decompression technique with preservation of a medial periorbital tissue strip is an adequate alternative technique in the therapy of optic neuropathy and exophthalmus reduction in endocrine orbitopathy with a low risk of postoperative motility disorders. PMID- 12766825 TI - [Plastic reconstructive correction of persistent orofacial swelling and swelling of the eyelids in Melkersson-Rosenthal syndrome]. AB - BACKGROUND: The Melkersson-Rosenthal syndrome complex is characterised by asymmetric orofacial oedema, facial palsy, furrowed tongue (lingua plicata) and additional neurological symptoms. PATIENT: A 21-year old man presented with a bilateral asymmetric oedema of the eyelids which increased during the follow-up period of 12 years. Intermittent swelling of the auditory channel occurred and resulted in hearing loss. Furthermore, the patient complained unspecific neurological symptoms as headache, tinnitus and subjective visual defects. The lid oedema did not only result in cosmetic problems but seriously restricted visual fields. RESULTS: Our patient showed the typical symptoms of the Melkersson Rosenthal complex, lacking however facial palsy which is often caused by mechanical compression along the course of the facial nerve. The differential diagnosis of recurrent swelling of the lids was discussed on an interdisciplinary plane. The inflammatory activity could be reduced and the frequency of swelling episodes could be diminished by a treatment with hydroxychloroquine. The initially relapsing, then persistent swelling had led to severe restriction of the visual field and resulted in a severe social stigmatisation and psychic stress for the patient. The surgical resection of the granulomatous tissue was carried out under steroid treatment to prevent recurrence. Histochemical analysis showed inflammation of the lymphoid plasmacellular type with mucipolysaccharidoid deposition typical of Melkersson-Rosenthal syndrome. Blepharoplasty resulted in an improved cosmetic appearance. The patient remained symptom-free with an excellent functional result. CONCLUSIONS: After persistence and constancy of the findings of a granulomatous blepharitis, surgical correction is a good opportunity to minimise the subjective complaints and neurological symptoms. PMID- 12766826 TI - [Long-term follow-up of bilateral endogenous Klebsiella endophthalmitis]. AB - BACKGROUND: Endophthalmitis subsequent to Klebsiella sepsis leads to functional blindness in most cases and is very difficult to treat. Every successful therapeutic modality can therefore help in creating an optimal therapeutic plan. CASE REPORT: A 69-year old diabetic patient exhibited bilateral Klebsiella endophthalmitis with sepsis after a pneumonia. Two intravenous antibiotics were used: aminoglycosides (Gentamycin) and cephalosporins (Cefotaxim or Cefuroxim) with local parabulbar injections of Prednisolon. The long-term follow-up of four years provided some overview of morphological aspects of the development of endophthalmitis. Characteristic greyish hypopyon was seen in both eyes, which was more pronounced in the left eye than in the right. The left eye became phthisic. After resorption of the hypopyon in the right eye and prolonged resorption of the subretinal abscess for 9 months a useful visual acuity at 0.2 was achieved. Two years after the endophthalmitis a cataract surgery with implantation of a posterior chamber silicon lens was performed and good visual acuity (0.6) was achieved. After four years, the subretinal abscess left an extremely large, sharp bordered, unpigmented scar up to the sclera. CONCLUSION: An early diagnosis and adequate long-time antibiotic therapy under the co-operative supervision of an ophthalmologist with internist appears to be most important for the therapeutic success in Klebsiella endophthalmitis. PMID- 12766827 TI - [VIII: Statistics in medical publications: check lists for authors]. AB - The "Material and Methods" section of clinical research papers should reference major aspects concerning statistical planning and evaluation of the study design and the resulting clinical data. Particular focus is laid on the listing of methods for description and significance evaluation of the trial data, as well as on design-associated study determinants (randomisation and masking strategy, response rate evaluation, primary endpoints, power and sample size determination, multiple testing, software and methods for statistical analysis). Suggestions for the minimum content to be mentioned in a trial publication are listed in author check lists and are illustrated by means of the study synopsis of a randomised clinical trial on the rotation stability of two lens haptics. PMID- 12766828 TI - Review of common therapeutic options in the United States for the treatment of pediculosis capitis. AB - Numerous therapies are available in both over-the-counter and prescription formulations for the treatment of head lice infestation. We summarize treatment recommendations from published literature and from a recent meta-analysis from the Cochrane Database of Systematic Reviews that describe the efficacy, safety, and resistance patterns of monotherapies available in the United States. If treatment with pyrethrin or permethrin fails to eradicate the infestation, the treatment of choice is malathion. However, because of malathion's flammability, it should be a second-line therapy. Orally administered ivermectin has been reported to be efficacious, but it is not currently a US Food and Drug Administration-approved pediculicide. Alternative therapies are also discussed, in addition to indications for prophylaxis, resistance reporting, and the social impact of infestation. PMID- 12766829 TI - Effectiveness of estriol-containing vaginal pessaries and nitrofurantoin macrocrystal therapy in the prevention of recurrent urinary tract infection in postmenopausal women. AB - We compared the efficacy and safety of estriol-containing vaginal pessary use with those of oral nitrofurantoin macrocrystal (NM) therapy for preventing urinary tract infection (UTI) in postmenopausal women with recurrent UTI. Over a period of 9 months, 86 women received an estriol-containing vaginal pessary (0.5 mg estriol) twice weekly, and 85 women received NM (100 mg) once daily. We recorded 124 episodes of UTI in women who received estriol-releasing pessaries and 48 episodes of UTI in women treated with NM (P=.0003). Twenty-eight women (32.6%) who received estriol had no episodes of UTI versus 41 women (48.2%) in the NM group. There was a significant increase in the number of superficial cells in women who received estriol, whereas in the NM group, no such changes occurred. However, there was no change in the extent of Lactobacillus colonization and in the vaginal pH in women who received estriol. Use of an estriol-containing pessary is less effective than oral NM therapy in the prevention of bacteriuria in postmenopausal women because of its failure to restore the population of lactobacilli and to reduce the vaginal pH in these women. PMID- 12766830 TI - Outbreak of joint and soft-tissue infections associated with injections from a multiple-dose medication vial. AB - An outbreak of Staphylococcus aureus joint and soft-tissue infections occurred after therapeutic injections in an outpatient setting. A physician performed intra-articular or soft-tissue injections on 17 patients in August 2001, and 5 (29%) were subsequently hospitalized for infections at the site. S. aureus was isolated from 4 patients, and all 4 isolates were indistinguishable by pulsed field gel electrophoresis. Of 10 patients injected with lidocaine and triamcinolone, 5 (50%) developed infections, compared with 0 of 7 patients injected with triamcinolone only (P=.04). A multiple-dose vial (MDV) of lidocaine was likely contaminated with S. aureus. A possible contributing factor was refrigeration after the use of MDVs of lidocaine; the manufacturer recommends storage at room temperature. An in vitro study of S. aureus in MDVs of lidocaine revealed prolonged survival at refrigerator temperatures. This outbreak highlights the importance of strict attention to aseptic procedures and carefully following manufacturers' instructions when using MDVs. PMID- 12766832 TI - Impact of ring wearing on hand contamination and comparison of hand hygiene agents in a hospital. AB - We determined risk factors for hand contamination and compared the efficacy of 3 randomly allocated hand hygiene agents in a group of surgical intensive care unit nurses. We cultured samples of one of the subjects' hands before and samples of the other hand after hand hygiene was performed. Ring wearing was associated with 10-fold higher median skin organism counts; contamination with Staphylococcus aureus, gram-negative bacilli, or Candida species; and a stepwise increased risk of contamination with any transient organism as the number of rings worn increased (odds ratio [OR] for 1 ring worn, 2.6; OR for >1 ring worn, 4.6). Compared with use of plain soap and water, hand contamination with any transient organism was significantly less likely after use of an alcohol-based hand rub (OR, 0.3; 95% confidence interval [CI], 0.1-0.8) but not after use of a medicated hand wipe (OR, 0.9; 95% CI, 0.5-1.6). Ring wearing increased the frequency of hand contamination with potential nosocomial pathogens. Use of an alcohol-based hand rub resulted in significantly less frequent hand contamination. PMID- 12766831 TI - Failure of a chloroquine chemoprophylaxis program to adequately prevent malaria during pregnancy in Koupela District, Burkina Faso. AB - In West Africa, administration of chloroquine chemoprophylaxis during pregnancy is common, but little is known about its impact on Plasmodium falciparum infection during pregnancy. Therefore, cross-sectional studies in antenatal care clinics (ANCs) and delivery units (DUs) were conducted in Koupela District, Burkina Faso. Chloroquine chemoprophylaxis was reported by 69% of 597 pregnant women at ANCs and by 93% of 853 women in DUs. P. falciparum peripheral parasitemia was identified in 29% of women at both ANCs and DUs. Placental parasitemia was identified in 22% of delivering women and was strongly associated with low birth weight (LBW) (risk ratio [RR], 1.7; 95% confidence interval [CI], 1.2-2.4) and prematurity (RR, 2.9; 95% CI, 1.6-5.4). In multivariate analysis, use of chemoprophylaxis was not associated with a reduction in the prevalence of placental parasitemia, LBW, or prematurity. Despite the high reported chloroquine chemoprophylaxis coverage, peripheral and placental malaria rates remain high and are associated with known adverse outcomes during pregnancy, including maternal anemia, prematurity, and LBW. Alternative prevention strategies, such as use of insecticide-treated mosquito nets and intermittent preventive treatment with more effective antimalarials, are needed. PMID- 12766833 TI - Molecular typing of Bordetella pertussis isolates recovered from Belgian children and their household members. AB - Recently, a moderate increase in the prevalence of pertussis, possibly contracted from adults, has been observed among unvaccinated children. During a 3-year period, we prospectively enrolled 93 index patients with a polymerase chain reaction (PCR) and/or culture result positive for Bordetella pertussis. Among 63 household contacts of 28 index patients, PCR and culture for B. pertussis identified 25 B. pertussis-positive persons. Nineteen of 25 B. pertussis-positive household contacts were asymptomatic. Isolates were available from 10 families of both index patients and household contacts for molecular typing by pulsed-field gel electrophoresis (PFGE) and for genotyping of pertactin and pertussis toxin by sequence-specific PCR and sequencing. PFGE demonstrated homogeneity among the isolates recovered from within each family but heterogeneity among the isolates recovered from different families. B. pertussis isolates recovered from index patients and their household contacts were indistinguishable by molecular typing, demonstrating that identical strains can cause full pertussis disease in children and asymptomatic infection in adults and adolescents. PMID- 12766834 TI - The health and economic burden of genital warts in a set of private health plans in the United States. AB - We estimated the prevalence of and costs associated with genital warts among privately insured individuals from the perspective of a private health plan in the United States. Health care claims data were derived from a sample of 3,664,686 privately insured individuals. The database was limited to cases of disease for which an insurance claim was generated, with costs reflecting inpatient, outpatient, and pharmacy payments from all sources. We identified 5095 cases of genital warts (1.7 cases per 1000 person-years) billed through the health plans during 2000. The prevalences of and health plan costs associated with genital warts were highest among women aged 20-24 years (6.2 cases and $1692 in costs per 1000 person-years) and men aged 25-29 years (5.0 cases and $1717 in costs per 1000 person-years). On average, individual episodes of care for genital warts involved 3.1 physician visits and incurred costs of $436. These are the first age- and sex-specific estimates of the prevalence and cost of genital warts for a US health plan. PMID- 12766835 TI - Fluoroquinolone-associated tendinopathy: a critical review of the literature. AB - With the expanded use of fluoroquinolones for the treatment of community-acquired respiratory infections and reports of tendon injury linked to the use of these agents, we reviewed the literature to investigate the frequency and strength of this association. Ninety-eight case reports were available for review. The incidence of tendon injury associated with fluoroquinolone use is low in a healthy population but increases in patients who have renal dysfunction, who are undergoing hemodialysis, or who have received renal transplants. Pefloxacin and ciprofloxacin were most frequently implicated, but tendon injury was reported with most fluoroquinolones. The median duration of fluoroquinolone treatment before the onset of tendon injury was 8 days, although symptoms occurred as early as 2 hours after the first dose and as late as 6 months after treatment was stopped. Up to one-half of patients experienced tendon rupture, and almost one third received long-term corticosteroid therapy. Tendon injury associated with fluoroquinolone use is significant, and risk factors such as renal disease or concurrent corticosteroid use must be considered when these agents are prescribed. PMID- 12766836 TI - African tick bite fever in travelers to rural sub-Equatorial Africa. AB - To estimate the incidence of, identify risk factors for, and describe the clinical presentation of travel-associated African tick bite fever (ATBF), a rapidly emerging disease in travel medicine, we prospectively studied a cohort of 940 travelers to rural sub-Equatorial Africa. Diagnosis was based on suicide polymerase chain reaction and the detection of specific antibodies to Rickettia africae in serum samples by multiple-antigen microimmunofluorescence assay, Western blotting, and cross-adsorption assays. Thirty-eight travelers, 4.0% of the cohort and 26.6% of those reporting flulike symptoms, had ATBF diagnosed. More than 80% of the patients had fever, headache, and/or myalgia, whereas specific clinical features such as inoculation eschars, lymphadenitis, cutaneous rash, and aphthous stomatitis were seen in < or = 50% of patients. Game hunting, travel to southern Africa, and travel during November through April were found to be independent risk factors. Our study suggests that ATBF is not uncommon in travelers to rural sub-Saharan Africa and that many cases have a nonspecific presentation. PMID- 12766837 TI - Outcomes analysis of delayed antibiotic treatment for hospital-acquired Staphylococcus aureus bacteremia. AB - The objective of this study was to determine the effect of delayed therapy on morbidity and mortality associated with nosocomial Staphylococcus aureus bacteremia. The study included all episodes of S. aureus bacteremia that developed >2 days after hospital admission during 1999 to 2001. Classification and regression tree analysis (CART) was used to select the mortality breakpoint between early and delayed treatment. During the 25-month study period, 167 patients met the inclusion criteria. The breakpoint between delayed and early treatment derived using CART was 44.75 hours. On multivariate analysis, delayed treatment was found to be an independent predictor of infection-related mortality (odds ratio, 3.8; 95% confidence interval, 1.3-11.0; P=.01) and was associated with a longer hospital stay than was early treatment (20.2 days versus 14.3 days; P=.05). These findings support the notion that delay of therapy has deleterious effects on clinical outcomes, and efforts should be made to ensure that appropriate therapy is initiated promptly. PMID- 12766838 TI - The use of economic modeling to determine the hospital costs associated with nosocomial infections. AB - Hospital-associated infection is well recognized as a patient safety concern requiring preventive interventions. However, hospitals are closely monitoring expenditures and need accurate estimates of potential cost savings from such prevention programs. We used a retrospective cohort design and economic modeling to determine the excess cost from the hospital perspective for hospital associated infection in a random sample of adult medical patients. Study patients were classified as being not infected (n=139), having suspected infection (n=8), or having confirmed infection (n=17). Severity of illness and intensive unit care use were both independently associated with increased cost. After controlling for these confounding effects, we found an excess cost of $6767 for suspected infection and $15,275 for confirmed hospital-acquired infection. The economic model explained 56% of the total variability in cost among patients. Hospitals can use these data when evaluating potential cost savings from effective infection-control measures. PMID- 12766839 TI - The impact of antimicrobial resistance on health and economic outcomes. AB - Despite an increasing prevalence of antimicrobial-resistant pathogens, the health and economic impact of colonization and infection with these organisms has not been fully elucidated. We explore how antimicrobial resistance can affect patient outcomes by enhancing virulence, causing a delay in the administration of appropriate therapy, and limiting available therapy. Next, we examine the different perspectives held by hospitals, third-party payers, patients, and society on the impact of resistance. Finally, we review methodological issues in designing and assessing studies that address the clinical outcomes for patients infected or colonized with resistant pathogens, including adjustment for important confounding variables, control group selection, and the quantification of economic outcomes. PMID- 12766840 TI - Routine cycling of antimicrobial agents as an infection-control measure. AB - Antimicrobial cycling is the deliberate, scheduled removal and substitution of specific antimicrobials or classes of antimicrobials within an institutional environment (either hospital-wide or confined to specific units) to avoid or reverse the development of antimicrobial resistance. True antimicrobial cycling requires a return to the antimicrobial(s) that were first used. Testing of the hypothesis that cycling will result in a lower prevalence of resistance is ongoing, mostly occurs within intensive care units, and largely involves cycling regimens targeted for treatment of suspected gram-negative bacterial infections. Unfortunately, there has been insufficient study to determine whether any meaningful impact on resistance has occurred as a result of a cycling program. Mathematical models question the usefulness of cycling as an infection-control method. Published studies demonstrate that cycling may be one way to change prescribing practices by clinicians without sacrificing patient safety. However, optimizing antimicrobial use through traditional and novel methods (e.g., computer decision support) should not be abandoned. PMID- 12766841 TI - Caspofungin. AB - Caspofungin, the first inhibitor of fungal beta-1,3 glucan synthesis to receive approval by the United States Food and Drug Administration, is effective for the treatment of mucosal and invasive candidiasis and invasive aspergillosis. It is also active in vitro and in animal models against a number of other filamentous and dimorphic endemic fungi and in animal models of Pneumocystis carinii infection. In vitro studies and some animal studies almost always indicate an absence of antagonism when caspofungin is combined with azole or polyene antifungal agents. Caspofungin has an excellent safety profile. Caspofungin may prove to be useful in empirical therapy for suspected invasive fungal infections. Additional clinical trial data that expand our knowledge of the usefulness of caspofungin for these and other mycoses, including its administration in combination with other antifungal agents, is anticipated. Caspofungin is an important addition to the antifungal pharmacopoeia. PMID- 12766842 TI - Bioterrorism web site resources for infectious disease clinicians and epidemiologists. AB - Finding bioterrorism-related information on the World Wide Web can be laborious. We hope to help readers find such information more easily by summarizing essential information in a consistent framework. A panel of 7 Centers for Disease Control and Prevention reviewers identified Web sites and evaluated them for sponsorship, mission, content usefulness, online ease of use, and adherence to commonly accepted quality criteria. Of >100 potential sites identified, 81 were chosen for target content of interest, and 43 were selected for inclusion. The results were classified into general purpose/portal sites; biological agent information; laboratory, infection control, epidemiology, and mental health information; and emergency contact sources, news and updates, event preparedness resources, information for first-responder settings, clinical and public education materials, and research resources. Agents covered included anthrax, smallpox, plague, botulism, tularemia, and viral hemorrhagic fever. PMID- 12766843 TI - Pharmacokinetics and pharmacodynamics of nelfinavir administered twice or thrice daily to human immunodeficiency virus type 1-infected children. AB - We studied the pharmacokinetics and pharmacodynamics of nelfinavir administered 2 or 3 times per day to human immunodeficiency virus type 1 (HIV-1)-infected children receiving highly active antiretroviral therapy containing nelfinavir. The geometric mean trough concentrations of nelfinavir for the thrice- and twice daily regimens were 1.55 mg/L and 1.11 mg/L, respectively (P=not significant). Nelfinavir concentrations did not correlate with total daily dose, dose per kilogram of weight, age, weight, previous protease inhibitor (PI) experience, or CD4(+) cell percentage. In the 25 PI-naive children, the virus load reductions at 24 weeks of treatment with the twice- and thrice-daily regimens were comparable. A significantly higher percentage of children in the twice-daily group had a trough concentration of nelfinavir of less than the inhibitory concentration of 95% (P=.042). The decrease in the virus load at 24 weeks of treatment was not correlated with the trough concentration of nelfinavir. The variability of trough concentrations was extremely high, particularly among recipients of the twice daily regimen, resulting in a higher number of patients with subinhibitory concentrations of nelfinavir in this group. PMID- 12766844 TI - Management of severely immunocompromised human immunodeficiency virus type 1 infected African orphans with structured treatment interruption: another kind of salvage therapy. AB - Without an effective therapeutic human immunodeficiency virus (HIV)-1 vaccine, providing cost-effective, minimally toxic antiretroviral therapy to the many individuals already infected with HIV-1 is a global priority. Implementation of a structured treatment interruption (STI) regimen may be a promising intervention for HIV-1 infection. We adopted this approach out of necessity for 3 severely immunocompromised African children naive to therapy, and increases in the T lymphocyte count and improved quality and sustainability of life 2 years after initiation of the STI regimen were noted. PMID- 12766845 TI - Mucormycosis (zygomycosis) in a heart-kidney transplant recipient: recovery after posaconazole therapy. AB - We describe the case of a diabetic patient who developed a severe invasive fungal infection due to Rhizopus species postoperatively after a dual heart/kidney transplantation with subsequent intensive immunosuppressive therapy. No improvement was noted with amphotericin B (deoxycholate) therapy, but salvage treatment with the new azole antifungal posaconazole (200 mg orally 4 times daily) resulted in dramatic clinical improvement as early as 1 week after the initiation of therapy that continued through 23 weeks of treatment, with marked clinical, mycological, and radiological improvements and no adverse events. PMID- 12766846 TI - Clearance of 14-3-3 protein from cerebrospinal fluid heralds the resolution of bacterial meningitis. AB - The 14-3-3 protein, a cerebrospinal fluid (CSF) marker of neuronal damage that was recently adopted for the diagnosis of Creutzfeldt-Jakob disease, is also found in the CSF of patients with a variety of neurological disorders. We prospectively studied 12 consecutive patients with purulent bacterial meningitis and found that 14-3-3 protein was detected in all patients at admission to the hospital. All patients who recovered cleared 14-3-3 protein from the CSF before discharge from the hospital (this was the first CSF marker to clear), whereas those who died never cleared the protein. PMID- 12766847 TI - Breakthrough fungemia caused by azole-resistant Candida albicans in neutropenic patients with acute leukemia. PMID- 12766848 TI - Campylobacter fetus bacteremia in a patient with adult T cell leukemia. PMID- 12766849 TI - Cholelithiasis possibly induced by protease inhibitors in 3 patients. PMID- 12766850 TI - Differences in pharmacokinetics of amphotericin B lipid formulations despite clinical equivalence. PMID- 12766851 TI - Extending treatment for HCV infection to HIV-HCV coinfected individuals with psychiatric illness and drug dependence. PMID- 12766853 TI - Importance of antifungal prophylaxis in patients who received a nonmyeloablative allogeneic PBSC transplant. PMID- 12766855 TI - Vascular thrombosis and acute cytomegalovirus infection in immunocompetent patients: report of 2 cases and literature review. AB - Acute cytomegalovirus (CMV) infection in immunocompetent patients is common worldwide, with seroprevalence rates of 40%-100%, depending on the country, socioeconomic conditions, and the patient's age. Infection is most often asymptomatic, but acute cytomegalovirus infection is occasionally revealed by prolonged fever, cervical lymphadenitis, and arthralgia, and it is more rarely revealed by pneumonia, myocarditis, pericarditis, colitis, and hemolytic anemia. Here, we report 2 cases of acute CMV infection in nonimmunocompromised adults that were complicated by venous thrombosis with pulmonary embolism. We also review previously reported cases of vascular thrombosis and discuss the propensity of CMV to induce vascular damage with associated thrombosis. PMID- 12766856 TI - Natural and nosocomial infection in a patient with West Nile encephalitis and extrapyramidal movement disorders. AB - Since its first recognition in North America in 1999, West Nile virus (WNV) has spread rapidly across the continent, but in many communities, rapid diagnostic tests for detection of WNV infection are not fully available. We describe a patient with extrapyramidal movement disorders and changes in the basal ganglia noted on magnetic resonance images that are characteristic of other flavivirus encephalitides and may help in the recognition of patients with West Nile encephalitis. Detailed molecular analysis suggested that, although our patient received a blood transfusion infected with WNV, the virus that caused his initial infection and encephalitis was probably acquired naturally from a mosquito. PMID- 12766857 TI - Linezolid-resistant, vancomycin-resistant Enterococcus faecium infection in patients without prior exposure to linezolid. AB - We describe 2 patients without prior exposure to linezolid who were infected with closely related strains of linezolid- and vancomycin-resistant Enterococcus faecium (LRVREF) that may have been hospital acquired. Polymerase chain reaction amplification of the domain V region of the 23S ribosomal RNA gene demonstrated the presence of the G2576U mutation previously reported to be associated with linezolid resistance. Nosocomial transmission of LRVREF is an ominous sign and underscores the importance of meticulous infection-control measures. PMID- 12766858 TI - Serum sickness-like reaction possibly associated with meropenem use. AB - Serum sickness-like reactions most commonly occur secondary to drug administration. We describe a serum sickness-like reaction that was possibly associated with meropenem therapy. PMID- 12766859 TI - [Bone mineral density in patients with multiple sclerosis: the effects of interferon]. AB - INTRODUCTION: Patients with multiple sclerosis (MS) are at greater risk of suffering from osteoporosis and pathological fractures, and the use of corticoids together with immobilisation and vitamin D deficiency is one of the causes of low bone mass (BM). AIMS. Our aim was to evaluate the effect of interferon, a drug that has only recently been introduced in the treatment of the disease, on bone mineral density (BMD) and bone remodelling markers. PATIENTS AND METHODS: A total of 30 females and 18 males with MS were studied. A standardised case history report was examined, and determinations of ionic calcium, vitamin D, osteocalcin, iPTH and urinary deoxypyridinoline, together with calcaneus densimetry measurements using a DEXA densimeter were also performed. RESULTS: The females treated with interferon had a BMD similar to that of those who had only received corticoids. Yet the males treated with interferon had a BMD that was lower than that of those who had not been treated with this drug (0.484 0.104 g/cm2 compared to 0.631 0.143 g/cm2, p= 0.032) and the control group (0.484 0.104 g/cm2 compared to 0.581 0.102 g/cm2, p= 0.015). No differences were found in the bone remodelling parameters. CONCLUSIONS: Males treated with interferon present a decrease in BM, and results are paradoxical because interferon plays a part in regulating bone metabolism and inhibits the development of osteoclasts, the cells responsible for bone resorption. PMID- 12766860 TI - [Diagnostic value of the criteria of the International Headache Society in the differential diagnosis of primary headaches]. AB - INTRODUCTION: In this research we study primary headaches, which are currently considered to be a public health problem. We also examine the fact that an ever increasing number of people are affected and that in some cases such headaches can eventually be disabling because they bring about different degrees of functional deterioration, which leads to negative repercussions on occupational capacity and on the quality of life of the people who suffer them. AIMS. Our aim is to determine the diagnostic value of the criteria of the International Headache Society (IHS) in the differential diagnosis of tension type headaches (TH) and migraine (M) in patients who attend the Neurology outpatients department at the Dr. Luis Guada Lacau clinic. PATIENTS AND METHODS: 100 consecutive patients, over the period between March and August 2001, were studied and, through an interview, we applied the IHS criteria to determine the type of primary headache they suffered from; blind comparisons of results were conducted against the diagnosis provided by an expert. RESULTS: Findings reveal a high degree of sensitivity (84, 85, 92 and 89%) for migraine with aura (MA), migraine without aura (MWA), TH, M + TH, respectively, and a specificity of 92, 95, 95 and 96% for MA, MWA, TH, M + TH, respectively, which correlates with the values obtained for the positive and negative predictive values, and the positive and negative probability coefficient. CONCLUSIONS: The IHS criteria are of great value in establishing HT, MA or MWA diagnoses and in those in which TH and M are presented together, and can therefore be used without risk in medical practice, if they are applied correctly. We recommend the use of these criteria in the General medicine and the Neurology clinic to detect cases of primary headaches and classify them correctly, thus enabling a correct diagnosis and effective therapy to be established. PMID- 12766861 TI - [Effectiveness of flunarizine in the prophylaxis of migraine: a meta-analytical review of the literature]. AB - BACKGROUND: Migraine is a common and disabling health problem among young and middle aged adults. Flunarizine have been used as a prophylactic medication in its management for more than two decades. OBJECTIVE: The aim of the study is to systematically review the evidence obtained from randomized controlled trials about the efficacy and security of flunarizine versus placebo for the prevention of migraine in adults. MATERIALS AND METHODS: Electronic search were performed using the databases MEDLINE, EMBASE, Biosis, the Cochrane Library, Lilacs and others. Reference lists of retrieved studies, reviews and conference abstracts were used to found another articles. Additionally, the authors performed a handsearched in recognized journals related to migraine and neurological topics. Randomized, placebo controlled, double blind trials assessing the efficacy of flunarizine for preventing migraine were included. We evaluated the frequency of the attacks by comparing the mean frequency before and after the intervention for each group of treatment and then the two groups were compared. To evaluate the security of flunarizine we used included and excluded studies and open trials. RESULTS: Nine studies were obtained but only four met the inclusion criteria. The fixed effects model reported a reduction in the frequency of attacks by using flunarizine. The monthly difference was 0.55 attacks (CI 95%= 0.215 0.895; p= 0.002). Somnolence was the most frequent adverse effect in 20.5% of the subjects (n= 1,987). CONCLUSIONS: Based on a small number of trials, flunarizine, at a daily dose of 10 mg lightly reduces the frequency of migraine attacks. PMID- 12766862 TI - [Screening for congenital hypothyroidism, phenylketonuria, galactosemia and biotinidase deficiency in a sample of mentally retarded patients in the city of Havana]. AB - INTRODUCTION: Congenital hypothyroidism (CH), phenylketonuria (PKU), galactosemia (GAL) and biotinidase deficiency (BD) are innate errors in metabolism that share varying degrees of mental retardation (MR) as a common characteristic. AIMS. The aim of our study was to screen individuals with MR of unknown origin for CH, PKU, GAL and BD. PATIENTS AND METHODS: Venous blood samples were collected on SS 903 specimen collection paper from 55 individuals with MR of unspecific origin born within the period 1977 1997. CH diagnosis was performed through determination of total thyroxine (T4) and thyroid stimulating hormone (TSH), using the UMELISA T4 and neonatal TSH reagent kits, respectively, and the detection of PKU, GAL and BD was conducted by determining phenylalanine (Phe), total galactose (Gal) and biotinidase enzyme activity (Biot) using UMTEST PKU, GAL and BIOTINIDASA. RESULTS: The mean values obtained for the analytes that were evaluated were: 0.8 mUI of TSH/L of total blood (EEM: 0.2), 113.1 nmol of T4/L of serum (EEM: 5.4), 67.7 mol of Phe/L of total blood (EEM: 0.1), 0.1 mmol of Gal/L of total blood (EEM: 0.01), and Biot activity was normal in all cases. CONCLUSIONS: This study enabled us to determine the T4, TSH, Phe and Gal levels in a sample from the Cuban population with MR of unknown causation. In addition, slightly higher levels of T4 were found in children who had hyperkinesis PMID- 12766863 TI - [Monitoring dermatomal somatosensory evoked potentials at the ERB point, the cervical spinal cord and the cerebral cortex in the diagnosis of cervical radiculopathy]. AB - INTRODUCTION: Recording at various levels of the somatosensory pathway is often used in somatosensory evoked potentials to mixed nerve stimulation (SEP), but not in dermatomal somatosensory evoked potentials (DSEP) in which only the cortical potential is usually recorded. The aim of our study was to compare the recordings of upper limb DSEP at Erb point, cervical cord, and subcortical and cortical levels with SEP recordings in healthy subjects and patients with cervical radiculopathy. PATIENTS AND METHODS: 17 patients with clinical history, MRI and electromyography consistent with cervical radiculopathy and 17 healthy subjects were included. Median and ulnar nerves were stimulated at the wrist; and C6, C7 and C8 dermatomes at the 1st, 3rd and 5th fingers respectively. All the potentials obtained with SEP and DSEP were compared between controls and patients by t test for independent samples. We also used Pearson s correlation for height/latencies, weight/amplitude and age/peripheral nerve conduction velocity (PNCV). RESULTS: DSEP potentials were of similar morphology of those observed in SEP but had longer latencies and smaller amplitudes. We found a positive correlation between height and latencies, and a negative association of weight with amplitude of peripheral potential, and age/PNCV. No difference between controls and the neurological intact segments of patients was found. 13 patient had DSEP altered while only 5 of them had altered SEP recorded. The most common finding was prolongation of the conduction time of the segment N9 N13 on DSEP recordings. CONCLUSION: We found that it is possible to record and to identify all the potentials in DSEP as observed in the SEP. On cervical radiculopathy, DSEP with the present technique increase the sensitivity and give some additional and useful information regarding the extension and localization of the pathology. Besides, DSEP recording is a non invasive technique, non traumatic and well tolerated for our patients. PMID- 12766864 TI - [Evaluation of diffuse cerebral atrophy in patients with a history of traumatic brain injury and its relation to cognitive deterioration]. AB - INTRODUCTION: Diffuse damage secondary to traumatic brain injury (TBI) can be studied through volumetric analysis of several structures that are sensible to this kind of injury, such as corpus callosum, ventricular system, hippocampus, basal ganglia and the volume of cerebrospinal fluid spaces. OBJECTIVE: Our aim is to describe how closed head injury (CHI) occurred in early years produce diffuse damage, and how this damage affects general cognitive functioning at long term. PATIENTS AND METHODS: Initially the group of subjects was composed of 27 head injured children and adolescents following paediatric moderate to severe TBI. From this initial group we selected 15 patients without focal lesion, or in case of having suffered focal lesion, this was smaller than 2,600 mm3. These subjects were assessed by means of volumetric analysis of cerebrospinal fluid spaces, corpus callosum, hippocampus and caudate nucleus, comparing the results with a matched control group. We calculated the degree of general cognitive ability of these subjects through tests of intellectual, memory, frontal lobe and motor speed functioning. RESULTS: This study demonstrates that early CHI produce a volume decrease in all measured structures. Corpus callosum atrophy is the factor that better explains general cognitive impairment. CONCLUSIONS: Diffuse damage secondary to moderate to severe peadiatric TBI has long term effects on several cerebral structures and on cognitive performance. Corpus callosum atrophy is the best predictor for general cognitive impairment, compared with other affected structures. PMID- 12766865 TI - [Ischemic stroke secondary to post varicella angiopathy in a 3 year old male]. AB - INTRODUCTION: Ischemic cerebrovascular disease is an infrequent problem in the paediatric age, but one which we now understand better thanks to modern neuroimaging techniques. We know little about its aetiopathogenesis, which is very varied, and it has been reported as being associated with viral infections such as chicken pox. CASE REPORT: A male, aged 3 years, who presented sudden hemiparesis, facial paralysis and dysarthria, without any other accompanying neurological symptoms. The patient had suffered a bout of chicken pox 6 weeks earlier. Of the abnormal complementary explorations, the most notable were varicella positive serological tests and MRI and MR angiography that pointed to ischemic involvement of the cortex. The patient progressed very well and the clinical features had completely reverted at five weeks. CONCLUSION: Post varicella angiopathy is one of the acquired risk factors for an ischemic stroke, and has been claimed to account for up to a third of all strokes in infants. Neuroimaging techniques allow the topographic determination of the ischemic repercussions. Patients usually progress well and in most cases the clinical features completely revert within a short time. We conclude that chicken pox should be included in the vaccination schedule as soon as possible in order to prevent complications that, as in the case we have reported, can be very serious. PMID- 12766866 TI - [Transient paraparesis as a manifestation of left carotid stenosis]. AB - INTRODUCTION: Paraparesis caused by a cerebrovascular disorder is infrequent, although it is seen in infraction of both anterior cerebral arteries (ACA), in vertebrobasilar insufficiency syndromes or in infarction of the border zones of the anterior circulation. CASE REPORT: Male aged 52 years, right handed, with a history of high blood pressure, who visited because of two episodes of transient paraparesis that lasted 5 minutes and 15 hours. During the last episode the presence of paraparesis and left Babinski sign were observed. Complementary explorations conducted to test for medullar pathology were negative. Cranial MRI showed only bilateral lacunar infarctions in deep territories. Four months later, the patient presented an episode of motor aphasia and paresthesia of the right lower limb, which remitted spontaneously in 10 minutes. Echo Doppler exploration of the supra aortic trunks showed significant stenosis of the left internal carotid artery (LICA) and occlusion of the right artery (RICA). Arteriography of the supra aortic trunks revealed a 99% stenosis of the RICA and 95% of the LICA, with vascularisation of both ACA dependent on the LICA. A left carotid endarterectomy was performed and the patient has remained asymptomatic to date. CONCLUSION: In our patient, both ACA depended on the flow from the LICA. We therefore consider that the symptoms of transient paraparesis were secondary to the left carotid stenosis, either due to a haemodynamic or an artery to artery embolic mechanism. PMID- 12766867 TI - [Deferred Horner syndrome following thoracoplasty]. AB - INTRODUCTION: Horner syndrome (HS) involves an injury affecting the ocular sympathetic nerve, which gives rise to myosis, palpebral ptosis and enophthalmos, and is accompanied by hemifacial anhidrosis in its complete forms. Its extension means that its involvement can occur in different structures and as a result of different medical and surgical processes. CASE REPORT: We describe the case of two patients who developed a subacute form of HS without involvement of the sweating process and which was not accompanied by any other clinical features affecting the orbit, neck, brain, spinal cord or of a radicular nature. Both of them had been submitted to thoracoplasty as therapy for tuberculosis over 30 years earlier. The complementary studies that were conducted did not reveal involvement of the ocular sympathetic nerve anywhere other than in the pleura. CONCLUSIONS: The lesion would have been produced in the endothoracic fascia, where the cervical sympathetic chain is closely related to the apical pleura, and the physiopathological mechanism would be fibrosis of the aforementioned structures. Many reports have been published that describe the onset of HS as an acute complication following thoracic surgery, but its late development is infrequent. PMID- 12766868 TI - [Congenital bilateral opercular syndrome. Presentation of three cases]. AB - OBJECTIVE: To present three children with bilateral opercular syndrome and the associated pathology. CASE REPORTS: We describe three patients, two of them are foster children, who have language problems, of chewing, of swallowing as well as continuous dribbling. Moreover, two case showed seizures that started at early age and were resisting to the antiepileptic medication. Motor deficit was disclosed in other two cases. The three patients were studied of the clinic, EEG and image point of view. The opercular bilateral syndrome diagnostic was made by the three dimensional magnetic resonance (3DMR) findings in the three cases. The images revealed open and large sylvian fissures and thickened insular cortex as well as polymicrogyria. CONCLUSION: The opercular syndrome shows peculiar clinical symptoms, but the diagnosis is made by the 3DMR alterations. PMID- 12766870 TI - [Combination of new electrophysiological and imaging techniques in the study of primary visual cortex function]. AB - INTRODUCTION: This work summarizes current research focused on explaining orientation selectivity of primary visual cortex (V1), and describes the electrophysiological and imaging techniques than are being used. DEVELOPMENT: The study of orientation selectivity in V1 is key to understanding the cortical mechanisms implicated in the processing of sensory information, but this enterprise has proved more challenging than previously thought and there is no consensus about the best model to explain V1 neurons activity. Ongoing research is focused on determining the importance of the different inputs that a cortical cell receives (thalamic and lateral cortical inputs), and their link to cortical architecture. To achieve that, current research is combining optical imaging techniques with intracellular recordings of V1 neurons. Recent findings have found differences in the synaptic integration performed by neurons located in the iso orientation domains vs orientation centers of the functional V1 map. CONCLUSIONS: Data describing synaptic activity combined with the cortical functional structure are yielding new clues about V1 computation, suggesting that there is more than one mechanism capable of generating orientation selectivity. PMID- 12766869 TI - [When should a brain scan with HMPAO be performed to diagnose brain death?]. AB - INTRODUCTION: Current Spanish law defines the criteria to be met in the case of brain death (BD) and the complementary tests to confirm it, although they do not always need to be carried out. In Spain one of the most frequently used methods is the electroencephalogram (EEG), which has its shortcomings in patients who are under sedation with barbiturates. Transcranial Doppler (TCD) offers a number of advantages in the diagnosis of BD, but it also has certain limitations. In these cases, a cerebral perfusion scan (CPS) with HMPAO can be very useful. CASE REPORTS: We describe the case of two patients who were admitted to the Intensive Care Unit with serious traumatic brain injuries. Progress was unfavourable and the findings of the clinical exploration were compatible with BD. Since we were dealing with patients under barbiturate therapy, the EEGs were complex and difficult to interpret. TCD was difficult to perform in both cases because of loss of brain matter and displacement of intracranial structures, respectively. CPS, however, allowed us to confirm the diagnosis of BD. CONCLUSION: Although it has obvious advantages, TCD cannot always be performed to confirm BD diagnosis. In these cases, CPS can be an extremely valuable tool PMID- 12766871 TI - [Neurological syndromes associated with the ingestion of plants and fungi with a toxic component (II). Hallucinogenic fungi and plants, mycotoxins and medicinal herbs]. AB - INTRODUCTION: A wide range of fungi and medicinal herbs, rich in hallucinogenic substances and widely used for mystic and medicinal purposes, can give rise to neurotoxic symptoms. DEVELOPMENT: We review the toxic syndromes that can arise from the ingestion of hallucinogenic fungi, cacti and plants, together with descriptions of cases of acute poisoning resulting from the use of medicinal herbs and from foodstuffs that are contaminated by mycotoxins. A series of different psychedelic fungi belonging to the Psilocybe, Panaeolus and Stropharia genera contain hallucinogenic alkaloids such as psilocybin. Some of the most notable plants displaying hallucinogenic and sedative properties are Papaver somniferum, Erytroxylum sp. and Cannabis sativa. Infusions of ayahuasca are obtained from the lianas and roots of different plants with psychoactive properties, such as Banisteriopsis caapi and Psychotria viridis, which contain alkaloids derived from tryptamine and from the beta carboline harmala. Peyote, a cactus rich in mescaline, and Claviceps purpurea (a fungus rich in LSD) are strong hallucinogens. We also examine ergotism and mycotoxicosis from Arthrinium sp. Poisoning from mycotoxin containing moulds on sugar cane can give rise to encephalopathy and late dystonia. Some of the more noteworthy medicinal plants for which neurological toxicity has been reported are Hypericum perforatum, kava kava (Piper methysticum), Aconitum sp. and Callilepis laureola. CONCLUSIONS: Because of the increasingly more widespread consumption of herbs and fungi and their potentially neurotoxic effects, in clinical practice there is a need to be aware of the neurological syndromes deriving from their use. PMID- 12766872 TI - [Data base for patients with balance disorders]. AB - INTRODUCTION: With the aim of integrating clinical thinking and obtaining epidemiological and biostatistical results in long series of patients who present balance disorders, we have developed a neuro otological protocol and its corresponding database. This offers answers to the questions of how to assess the patient, how to conduct a clinical interview correctly, and which complementary studies should be requested and how. RESULTS: The information from the protocol is stored and analysed in our database, which was designed in the Epi Info software application, produced by CDC (NIH, USA), in collaboration with the WHO Global Program on AIDS. The application, which we call ENO LK, has already been used to store data concerning 1,100 patients whose average age is 54.5 years old and 62.8% of which were females (SD 18, range 4 93). 69.1% were diagnosed as suffering from vertigo, 12.7% displayed instability, 1.9% syncope and 16.3% had other causes (37% psychogenic and 28% disorders affecting the central integrator). Of the 760 patients with vertigo, in 55% it was positional (60% of these were idiopathic benign), 6.3% were sustained (peripheral causes accounted for 74% and a vascular aetiology was predominant in the central causes), 26.6% were recurring and 12.1% otolithic (in this series the vertigos all had a central aetiology). CONCLUSIONS: With this application the user has the possibility of obtaining epidemiological and diagnostic conclusions efficiently and effectively, as well as aiding to follow up all patients who present balance disorders. PMID- 12766873 TI - [Verbal communication disorders and language disorders in cortical dementias (I). The current state of the art in research]. AB - INTRODUCTION: This is the first part of a research work into primary and secondary language disorders (LD) in the stages of minimum to mild pathological deterioration in the degenerative process of cortical dementias. AIMS. In this first part, in the light of recent models of symbolic computation of language processing, we review the most significant publications on the subject matter. DEVELOPMENT: We begin with an introduction to the historical development of the approaches used to deal with the subject. Following that, we briefly describe the components of the language processing system (LPS) in the light of symbolic computation models. We then analyse the publications dealing with those models. Our aim was to determine whether the LD reported in those publications are the result of a primary involvement of any of the LPS components, or whether they are only the result of disorders affecting other components of the cognitive system. In this case, the supposed LD would be secondary and thus a communication, but not language, disorder. CONCLUSIONS: Despite the important variations from one individual to another that have been systematically observed by different authors, it seems that only LD belonging to so called primary progressive aphasia are actually cases of this disorder. In all the other dementias in the above mentioned stages of the development process these disorders are secondary to the involvement of other subsystems of the LPS. PMID- 12766874 TI - [Interferon beta in multiple sclerosis]. AB - INTRODUCTION AND METHOD: The mechanism of action of interferons We do not know in the multiple sclerosis but, although exist discrepancies in relation to their degree of effectiveness, fundamentally the dose to use and the administration route, the three available ones have proven to be effective in the reduction of the number of the bouts. We reviewed the more important clinical tests made with three interferons in the remitent recurrent (RR) forms, the isolated neurologic syndromes and the secondary progressive (SP) forms of the disease and reviewed the comparative studies between interferons. CONCLUSIONS: The three interferons are well tolerated although the intramuscular form produces less skin effects; in the RR forms the number of bouts are reduced in significant form. Only the study with 1a intramuscular was demonstrated effectiveness in the SP forms and with the subcutaneous IFN b 1b exists discrepancies between the European and the American studies. The early treatment with subcutaneous IFN b 1a slow down the evolution of the disease. Except in relation to the side effects, there are not evidence that the dose and the administration route are important in the effectiveness in the long term. PMID- 12766875 TI - [Peripheral facial paralysis. The value of clinical neurophysiology]. AB - AIMS: The aim of this study was to review the value of neurophysiological exploration in peripheral facial paralysis in the literature and in our own experience. METHOD: Peripheral facial paralysis is a frequently occurring cranial neuropathy with an acute presentation. Its striking clinical expression is usually in strong contrast to its benign prognosis, except for a small percentage of cases in which the existence of acute injury to the nerve (axonotmesis) will cause it to progress unfavourably, either due to the absence of nerve regeneration or because this is inadequate or insufficient. A neurophysiological study aids diagnosis and the functional and prognostic assessment. CONCLUSIONS: We recommend the utilisation of standardised studies involving a comprehensive electromyographic and electroneurographic evaluation of the facial territory, including reflexology (blink reflex). Use of a suitable methodology in the neurophysiological study of this neuropathy will allow us to complete the diagnosis and to evaluate progress, even from very early days (early surgical approach prior to the onset of nerve degeneration), all of which is decisive in the prognosis of injury throughout the process. PMID- 12766876 TI - [Eagle's syndrome: findings from computerised tomography scans]. PMID- 12766877 TI - [Pneumocephalus following epidural anaesthesia]. PMID- 12766878 TI - Antigen-specific immune function after hematopoietic stem cell transplantation. AB - Hematopoietic stem cell recipients are characterized by an immunodeficiency of varying severity and duration. The present review focuses on the antigen-specific function of recipients with the hypothesis that the acquisition of antigen specific function is predictive of the recipient's capacity to resist lethal infection with environmental pathogens. PMID- 12766879 TI - Role of tumor necrosis factor-alpha in graft-versus-host disease and graft-versus leukemia responses. AB - Tumor necrosis factor-alpha (TNF-alpha) antagonist therapy has proven effective in inflammatory conditions such as rheumatoid arthritis and Crohn's disease. There is substantial evidence that TNF-alpha also plays a role in the development of graft-versus-host disease (GVHD) after allogeneic hematopoietic cell transplantation, which along with leukemia relapse remains one of the 2 major impediments to success of the approach. Using a recently developed potent rat/mouse chimeric monoclonal antibody directed against murine TNF-alpha (CNTO2213), the authors investigated the effect of TNF-alpha blockade on GVHD mediated by either CD4(+) or CD8(+) donor T cells. The results indicated that the treatment had only a moderate effect on both a CD8(+) T cell-mediated major histocompatibility complex-matched GVHD model involving multiple minor histocompatibility antigens and a p-->F(1) acute GVHD model directed against a haplo-mismatched major histocompatibility complex barrier involving both CD4(+) and CD8(+) T cells. In contrast, treatment with the anti-TNF-alpha antibody had a highly significant effect (100% survival rate) on the CD4(+) T cell-mediated component of this latter model. Importantly, anti-TNF-alpha antibody did not block the development of a graft-versus-leukemia effect against a murine myeloid leukemia challenge in either a syngeneic or allogeneic p-->F(1) setting. This suggests that the inhibition of TNF-alpha during allogeneic hematopoietic cell transplantation may be able to diminish the inflammatory GVHD reaction without hindering effective graft-versus-leukemia responses. PMID- 12766880 TI - Posttransplant day significantly influences pharmacokinetics of cyclosporine after hematopoietic stem cell transplantation. AB - Cyclosporine-based immunosuppression is common after allogeneic hematopoietic stem cell transplantation (HSCT). Elevated cyclosporine concentrations are associated with significant toxicity and often result in the temporary cessation or discontinuation of cyclosporine. Low blood concentrations also result in significant immunologic risks, primarily graft-versus-host disease and loss of stem cell graft. The pharmacokinetics of cyclosporine are highly complex, and maintaining therapeutic and safe cyclosporine concentrations are challenging. Several clinical factors are known to independently influence in vivo cyclosporine pharmacokinetic behavior. However, in the critically ill patient, several of these clinical factors are generally present simultaneously. Unfortunately, there are no studies that have evaluated the combined effects of these clinical factors on cyclosporine disposition in HSCT. The objective of our study is to determine the population pharmacokinetic parameters of intravenous and oral cyclosporine, evaluate the effects of clinical covariates on cyclosporine pharmacokinetics, and develop a model that estimates clearance (Cl) and dose requirements for an individual HSCT patient with these clinical covariates. The authors analyzed 740 cyclosporine steady-state whole blood concentrations in 129 adult patients obtained between day 0 and discharge or 60 days posttransplant, whichever came first. Patients received intravenous cyclosporine at 2.5 mg/kg every 12 hours if body weight was greater than 50 kg, 2.5 mg/kg every 8 hours if less than 50 kg, or 5 to 7.5 mg/kg/d given as a continuous infusion, beginning on day-3. Patients were converted to oral therapy as tolerated. The influence of clinical covariates on the Cl of cyclosporine was tested with a nonlinear mixed effects model (NONMEM). The tested clinical covariates were age, height, body weight on admission, body surface area, sex, type of hematologic malignancy, transplant type, preparative regimen, baseline serum creatinine, T-cell depletion of graft, number of methotrexate doses, day of onset, and maximum grade of acute graft-versus-host disease. The route and frequency of cyclosporine administration, day posttransplant, total bilirubin level, serum creatinine level, actual body weight, presence of concurrent CYP450 enzyme inhibitors and inducers, or nephrotoxins on the day of the cyclosporine blood measurement were also evaluated. Cyclosporine Cl significantly decreased each week posttransplant. The authors found no significant effect of any of the other tested covariates including total bilirubin on Cl. The final regression model for the estimation of Cl is: Cl (L/hr) = ([body weight in kg - 70] * 0.183 + 22.3) * (day posttransplant factor). The corresponding day posttransplant factor estimates are 1.46, 1.32, 1.20, and 1.0 during days 0 to 7, 8 to 14, 15 to 21 and greater than 21 posttransplant, respectively. The interindividual variability in Cl was 27.7%. The dose of intravenous or oral cyclosporine can be calculated using the estimated Cl. Understanding cyclosporine pharmacokinetics and the clinical events that lead to alterations in Cl and exposure is critical in optimizing immunosuppressive therapy. The authors found that cyclosporine Cl significantly decreased posttransplant until day 21. A pharmacokinetics model was developed that incorporates the day posttransplant to predict cyclosporine Cl. Cyclosporine dose requirements in an individual HSCT patient to achieve the desired therapeutic blood target can be estimated using this model. PMID- 12766881 TI - Long-term follow-up of T cell-depleted allogeneic bone marrow transplantation in refractory multiple myeloma: importance of allogeneic T cells. AB - Multiple myeloma may be cured by myeloablative conditioning and allogeneic blood or marrow transplantation (alloBMT), but this occurs at the expense of high transplant-related mortality. In an endeavor to reduce procedure-related toxicity, this study retrospectively evaluated the safety, tolerability, and efficacy of T cell depletion by counterflow centrifugal elutriation before alloBMT. Fifty-one patients with stage II (6) or III (45) multiple myeloma received alloBMTs using T cell depletion by elutriation. Fifty-three percent (27 of 51) of patients had primary refractory disease at the time of transplantation, 10% (5 of 51) had relapsed disease, and 4% (2 of 51) had refractory relapsed disease. The median age was 49 (range, 32 to 62) years, and the median time from diagnosis to transplantation was 9 (range, 4 to 58) months. Patients had received a median of 1 (range, 1 to 3) regimen and 4 (range, 2 to 16) cycles of chemotherapy. In this population, transplant-related mortality rate was 24% (12 of 51) with 2 patients dying of graft-versus-host disease (GVHD). Thirty-one of 39 evaluable patients have experienced relapse, and the probability of progression-free survival 5 years after alloBMT alone is 16%. Sixteen patients were given donor lymphocyte infusions (DLI) at the time of relapse (n = 11) or for persistent disease 1 year after transplantation (n = 5). Acute or chronic GVHD was seen in 63% (10 of 16) of patients given DLI. Responses were seen in 8 of 16 patients (6 complete response [CR], 2 partial response [PR]) with 6 of 8 responding patients having GVHD. Five recipients of DLI remain in a continuous CR, ranging from 3 to 64 months in duration. Thus, like chronic myelogenous leukemia, allogeneic T cells appear to have potent antimyeloma activity that is critical for achieving a cure. DLI-induced remissions of multiple myeloma can be durable, even in patients with refractory multiple myeloma. Unlike chronic myelogenous leukemia, the antimyeloma effect of allogeneic T cells rarely occurs in the absence of clinically significant GVHD. PMID- 12766882 TI - Impact of prophylactic donor leukocyte infusions on mixed chimerism, graft-versus host disease, and antitumor response in patients with advanced hematologic malignancies treated with nonmyeloablative conditioning and allogeneic bone marrow transplantation. AB - In an attempt to capture graft-versus-tumor effects without graft-versus-host disease (GVHD), the authors initiated a trial of nonmyeloablative allogeneic bone marrow transplantation (BMT) in patients with advanced hematologic malignancies, with the majority of patients having chemotherapy-refractory disease. Forty-two patients received an HLA-matched related donor BMT after a cyclophosphamide and antithymocyte globulin-based conditioning that also included thymic irradiation for patients who had not received prior mediastinal radiotherapy. Prophylactic donor leukocyte infusion (pDLI) at a dose of 1 x 10(7) CD3(+) cells per kilogram were given beginning 5 weeks post-BMT to 16 patients with mixed chimerism (MC) but without GVHD, whereas 26 patients did not receive pDLI, either because of GVHD or early relapse. Twelve of 16 patients (75%) receiving pDLI had T cell chimerism at the time of pDLI >/=40%. These patients, by day 100 post-BMT, either converted to full donor chimerism (FDC) (n = 10) or had an increase in or stable donor chimerism (n = 2) after pDLI. Four of 4 patients whose T cell chimerism was /=grade II acute GVHD, including 12 after BMT and 7 after pDLI. Approximately one third of patients, after having initial MC, eventually lost their donor graft. The authors conclude that (1) pDLI has the potential to convert MC to FDC; (2) sustained remissions can be achieved in patients with chemorefractory hematologic malignancies who receive a pDLI, albeit with a significant risk of acute GVHD; and (3) the degree of donor T cell chimerism at the time of pDLI is predictive of the fate of MC, ie, donor T cell chimerism >/=40% or /=50% reduction in the Swenerton score during SDCT (odds ratio [OR] 3.32, P <.01) and soft-tissue disease (OR 4.08, P <.01). The presence of multiple metastatic sites predicted decreased probability of conversion to CR (OR 0.34, P <.01). The Swenerton score provides a thorough estimate of disease extent, and reduction of Swenerton score by SDCT is potentially useful for selecting the optimal candidates for HDCT trials who may achieve long-term disease control. PMID- 12766884 TI - Adenovirus infection in hematopoietic stem cell transplantation: effect of ganciclovir and impact on survival. AB - Adenoviruses (ADV) are emerging as important causes of morbidity and mortality in patients undergoing hematopoietic stem cell transplantation (HSCT). In mainly non T-cell depleted HSCT recipients, we analyzed the incidence of ADV infection, risk factors for infection, the effect of ganciclovir administered for prevention of cytomegalovirus (CMV), and the impact of ADV infection on survival. The overall incidence of ADV, irrespective of the method of detection, was 8.5% (450/5233) and 12.3% (43/348) after the first or second allogeneic HSCT, and 6.3% (78/1219) and 6.5% (5/77) after the first or second autologous HSCT, respectively. The most frequent sites of infection and disease were stool and gastrointestinal tract, respectively. Statistically significant risk factors associated with ADV infections among allogeneic recipients included younger age, grade II to IV graft versus-host disease, year of transplantation, and a second allogeneic HSCT. Furthermore, allogeneic patients seronegative for CMV at transplantation and seropositive allogeneic patients who did not receive ganciclovir, either at engraftment or as pre-emptive therapy on CMV reactivation, were at higher risk of developing ADV infections compared with seropositive patients who received ganciclovir (odds ratio=1.8, 95% confidence interval (CI) 1.2 to 2.8, P=.005 and odds ratio=3.4, 95% CI 2.1 to 5.55, P<.0001, respectively). The hazard of overall mortality was higher in patients who contracted ADV compared with those who did not (hazard ratio 1.5, 95% CI 1.3 to 1.7, P<.0001). This study shows that ADV infections are associated with poor transplantation outcome in T-cell repleted HSCT recipients. Ganciclovir, given for CMV prevention, may have a protective effect. Controlled treatment and prevention studies are warranted. PMID- 12766890 TI - A multi-center 1H MRS study of the AIDS dementia complex: validation and preliminary analysis. AB - PURPOSE: To demonstrate the technical feasibility and reliability of a multi center study characterizing regional levels of the brain metabolite ratios choline (Cho)/creatine (Cr) and myoinositol (MI)/Cr, markers of glial cell activity, and N-acetyl aspartate (NAA)/Cr, a marker of mature neurons, in subjects with AIDS dementia complex (ADC). MATERIALS AND METHODS: Using an automated protocol (GE PROBE-P), short echo time spectra (TE = 35 msec) were obtained at eight sites from uniformly prepared phantoms and from three brain regions (frontal white matter, basal ganglia, and parietal cortex) of normal volunteers and ADC and HIV-negative subjects. RESULTS: A random-effects model of the phantom and volunteer data showed no significant inter-site differences. Feasibility of a multi-center study was further validated by detection of significant differences between the metabolite ratios of ADC subjects and HIV negative controls. ADC subjects exhibited significantly higher Cho/Cr and MI/Cr in the basal ganglia and significantly reduced NAA/Cr and significantly higher MI/Cr in the frontal white matter. These results are consistent with the predominantly subcortical distribution of the pathologic abnormalities associated with ADC. CONCLUSION: This is the first study to ascertain and validate the reliability and reproducibility of a short echo time (1)H-MRS acquisition sequence from multiple brain regions in a multi-center setting. It should now be possible to examine the regional effects of HIV infection in the brain in a large number of subjects and to study the metabolic effects of new therapies for the treatment of ADC in a clinical trial setting. PMID- 12766891 TI - Proton spectroscopic metabolite signal relaxation times in preterm infants: a prerequisite for quantitative spectroscopy in infant brain. AB - PURPOSE: To determine relaxation times of metabolite signals in proton magnetic resonance (MR) spectra of immature brain, which allow a correction of relaxation that is necessary for a quantitative evaluation of spectra acquired with long TE. Proton MR spectra acquired with long TE allow a better definition of metabolites as N-acetyl aspartate (NAA) and lactate especially in children. MATERIALS AND METHODS: Relaxation times were determined in the basal ganglia of 84 prematurely born infants at a postconceptional age of 37.8 +/- 2.2 (mean +/- SD) weeks. Metabolite resonances were investigated using the double-spin-echo volume selection method (PRESS) at 1.5 T. T1 was determined from intensity ratios of signals obtained with TRs of 1884 and 6000 msec, measured at 3 TEs (25 msec, 136 msec, 272 msec). T2 was determined from signal intensity ratios obtained with TEs of 136 msec and 272 msec, measured at 2 TR. Taking only long TEs reduced baseline distortions by macromolecules and lipids. For myo-inositol (MI), an apparent T2 for short TE was determined from the ratio of signals obtained with TE = 25 msec and 136 msec. Intensities were determined by fitting a Lorentzian to the resonance, and by integration. RESULTS: Relaxation times were as follows: trimethylamine-containing compounds (Cho): T1 = 1217 msec/T2 = 273 msec; total creatine (Cr) at 3.9 ppm: 1010 msec/111 msec; Cr at 3.0 ppm: 1388 msec/224 msec; NAA: 1171 msec/499 msec; Lac: 1820 msec/1022 msec; MI: 1336 msec/173 msec; apparent T2 at short TE: 68 msec. CONCLUSION: T1 and T2 in the basal ganglia of premature infants do not differ much from previously published data from basal ganglia of older children and adults. T2 of Cho was lower than previous values. T2 of Cr at 3.9 ppm and Lac have been measured under different conditions before, and present values differ from these data. PMID- 12766892 TI - Regional dynamics of the fMRI-BOLD signal response to hypoxia-hypercapnia in the rat brain. AB - PURPOSE: To examine the regional blood oxygenation level-dependent (BOLD) signal response to rapid changes in arterial oxygen tension. MATERIALS AND METHODS: Functional MR imaging (fMRI) was carried out in five male Sprague-Dawley rats anesthetized with Sodium Pentobarbital. Rats were subjected to different durations of apnea as a rapid, graded, and reversible hypoxic-hypercapnic stimulus. Dynamics of the BOLD signal response were studied on a pixel-by-pixel basis in the cerebral cortex, hippocampus, third ventricle, and thalamus in the rat brain. RESULTS: Apnea induced a BOLD signal drop in all the brain regions studied, the magnitude of which increased with longer durations of the stimulus. The signal recovered to preapnic baseline levels after resumption of normal ventilation. Regional variation in the BOLD signal dynamics was observed with the magnitude of the BOLD signal change in the hippocampus being the least, followed by a relatively larger change in the thalamus, cerebral cortex, and third ventricle. The time (t(0)) for the signal change after the onset of the stimulus was estimated for every pixel. Time delay maps generated show the highest onset time values in the hippocampus followed by the thalamus, cerebral cortex, and third ventricle. CONCLUSION: The regional dynamics of the BOLD signal in the brain in response to apnea may vary depending on the rate of oxygen metabolism in addition to cerebral blood flow (CBF). PMID- 12766893 TI - Double inversion black-blood fast spin-echo imaging of the human heart: a comparison between 1.5T and 3.0T. AB - PURPOSE: To evaluate the effectiveness of blood suppression and the quality of black-blood cardiac images acquired at 3.0 Tesla using a double-inversion recovery fast spin-echo sequence by comparing data acquired at 3.0T to data acquired at 1.5T. MATERIALS AND METHODS: Black-blood T2-weighted fast spin-echo images of the heart were acquired from five normal volunteers at 1.5T and five normal volunteers at 3.0T. Region-of-interest signal intensity measurements were performed at several locations in the suppressed blood regions of the left and right ventricles and around the left ventricle walls to assess the effectiveness and uniformity of the blood suppression, the myocardial signal-to-noise ratio (SNR), and the signal uniformity at both field strengths. B1 field maps were produced in phantoms and in subjects at both field strengths. RESULTS: Blood suppression performance is equivalent at 1.5T and 3.0T. The improvement in SNR at 3.0T compared with 1.5T is less than has been predicted in previous studies. The signal uniformity is significantly poorer at 3.0T than at 1.5T due to dielectric effects and shorter radio frequency wavelengths (P < 0.005). CONCLUSION: Spin echo and spin-echo echo-train sequences that perform well at 1.5T will produce large signal variations in the chest cavity at 3.0T without modifications. B1 insensitive methods must be explored and implemented for spin-echo sequences to fully realize the advantages of using these sequences for high-field MRI. PMID- 12766894 TI - Improvement of image quality of non-invasive coronary artery imaging with magnetic resonance by the use of the intravascular contrast agent Clariscan (NC100150 injection) in patients with coronary artery disease. AB - PURPOSE: To assess the feasibility of Clariscan, an intravascular contrast agent, for free breathing, navigator assisted, high resolution, three-dimensional magnetic resonance coronary angiography (MRCA) in patients, as extracellular contrast agents are unfavorable for the improvement of image quality. MATERIALS AND METHODS: MRCA was performed in 10 patients with known coronary artery disease (CAD) with (1-5 mg Fe/kg body weight) and without contrast agent. RESULTS: Compared to unenhanced images, Clariscan did not improve signal-to-noise (SNR) or contrast-to-noise ratios (CNR) compared to fat or myocardium in the proximal parts of the coronary arteries. However, when analyzing the peripheral parts (>4 cm from origin), CNR(fat) and CNR(myo) improved up to a factor of 1.81 and 5.85, respectively, at a dose of 3 mg Fe/kg body weight, while SNR did not reach statistical significance. The visible length of the coronary arteries was improved from 49 +/- 18 mm to 73 +/- 33 mm. The proximal diameter was reduced from 3.6 +/- 0.8mm to 3.2 +/- 0.8 mm, representing more closely the diameter of 3.1 +/- 0.7 mm measured by quantitative coronary angiography. Of 11 significant stenoses (>50%), eight were identified in the enhanced compared to six in the unenhanced images. CONCLUSION: The use of Clariscan at a dose of 2-3 mg Fe/kg body weight improves image quality of three-dimensional-MRCA, especially in the peripheral segments, and, thus, the diagnostic accuracy for the detection of CAD. PMID- 12766895 TI - Laser-induced thermotherapy (LITT) of liver metastases: MR-guided percutaneous insertion of an MRI-compatible irrigated microcatheter system using a closed high field unit. AB - PURPOSE: To evaluate the efficacy and safety of a new MRI-compatible irrigated laser microcatheter system for thermal ablation of liver metastases. MATERIAL AND METHODS: The new microcatheter system consists of a titanium needle with a diameter of 1.5 mm and a surrounding Teflon catheter with an outer diameter of 1.8 mm (5.5 F). In vitro laser-induced coagulation of bovine liver tissue was performed to determine the optimal perfusion rate of cooling saline flow, maximum laser energy, and ablation time. Laser-induced thermotherapy using the new microcatheter system, an Nd:YAG laser (Dornier), and a flexible laser light guide (Somatex GmbH, Berlin, Germany) was performed in 28 patients with liver metastases. Percutaneous insertion and positioning of multiple microcatheters in the lesion and monitoring of therapy was performed with a closed high-field MRI scanner using T1-weighted gradient-echo sequences during breath-hold. RESULTS: A perfusion rate of 0.75 mL/minute, a laser energy of 15 W, and an ablation time of 20 minutes were found suitable to achieve safe and sufficient ablation of metastatic tissue. The mean volume of induced coagulation in vitro was 23.9 mL. Ablation of liver metastases in patients was technically and clinically successful. CONCLUSION: The new microcatheter system allows for both catheter placement and monitoring of therapy using a single imaging modality. This shortens the procedure and enables more precise puncture and therapy of liver metastases. Due to the miniaturized design of the catheter and the real-time monitoring, the procedure is minimally invasive and very well tolerated by patients. This new technique seems to be a safe and feasible alternative in treating liver metastases. PMID- 12766897 TI - Visualization and quantification of renal R2* changes during water diuresis. AB - PURPOSE: To refine and evaluate methods for analysis of renal blood oxygenation level dependent (BOLD) MRI data. MATERIALS AND METHODS: Color R2* maps and regions-of-interest (ROIs) on the borderline between cortex and medulla were applied to renal BOLD MRI data of a group of 13 young female subjects. RESULTS: The distribution of R2* within the kidneys was heterogeneous and the response of human kidneys to water diuresis was patchy. R2* values at the cortico-medullary border region have a smaller variation than in wider cortical or medullary regions and are sensitive to physiological changes produced by water diuresis. CONCLUSION: These methods provide improved visualization of the regional distribution of R2* and its variations and more precise quantification of the changes in renal R2* produced by water diuresis. PMID- 12766896 TI - Kidneys in hypertensive rats show reduced response to nitric oxide synthase inhibition as evaluated by BOLD MRI. AB - PURPOSE: To examine whether the noninvasive technique of blood oxygenation level dependent magnetic resonance imaging (BOLD MRI) can detect changes in renal medullary oxygenation following administration of a nitric oxide (NO) synthase inhibitor, N(G)-nitro-L-arginine methyl ester (L-NAME). Hypertension is associated with endothelial dysfunction and is characterized by a lack of response to endothelial-dependent vasoactive substances, including nitric oxide synthase inhibitors. We hypothesized that the magnitude of the change would be reduced in the kidneys of hypertensive subjects relative to normal controls. MATERIALS AND METHODS: To test this hypothesis, data were obtained in spontaneously hypertensive rats (SHR, n = 6). Wistar-Kyoto rats (WKY, n = 7) were used as normotensive controls. RESULTS: As expected, WKY rats showed a significant response to L-NAME (R(2)* increasing from 23.6+/-1.5 Hz to 32.5+/-2.2 Hz, P < 0.05), while SHR exhibited a minimal change in medullary oxygenation (R(2)* measuring 31.9+/-2.8 Hz pre- and 35.5+/-2.2 Hz post-L-NAME). The baseline R(2)* in SHR is found to be comparable to post-L-NAME values in WKY rats, suggesting a basal deficiency of nitric oxide in SHR. CONCLUSION: Based on the differential effect of NO synthase inhibition on medullary oxygenation, BOLD MRI can distinguish hypertensive from normal kidney. Our results are consistent with previously reported observations using invasive methods. PMID- 12766898 TI - Functional evaluation of normothermic ischemia and reperfusion injury in dog kidney by combining MR diffusion-weighted imaging and Gd-DTPA enhanced first-pass perfusion. AB - PURPOSE: To evaluate functional alterations of renal ischemia and reperfusion injury using MR diffusion-weighted imaging and dynamic perfusion imaging. MATERIALS AND METHODS: Twelve dogs were randomly divided into four groups. Animal renal ischemia was respectively induced for 30 (group 1), 60 (group 2), 90 (group 3), and 120 (group 4) minutes by left renal artery ligation under anesthesia. Using a 1.5 T MR system, true-FISP, TSE, EPI, and DWI sequences were acquired in five different periods; specifically, pre-ischemia, onset-ischemia, late ischemia, onset-reperfusion, and post-reperfusion. Moreover, a turbo-FLASH sequence (TR/TE/TI/FA = 5.8/3.2/400 msec/10 degrees ) with a temporal resolution of 1.16 seconds was acquired. Signal intensity (SI) was measured in the cortex, outer medulla, and inner medulla of kidney. Apparent diffusion coefficient (ADC) values were calculated, and SI was plotted as a function of time. RESULTS: In all animals, significant SI changes of the left kidney on T2/T2*WI were detected following ischemia-reperfusion insult compared to corresponding values of the right kidney. Following ligation, the ADC values decreased in all layers of the left kidney. Immediately after the release of ligation, ADC values in both outer and inner medulla of the left kidney remained lower than those of the right kidney in those animals which were induced with renal ischemia for 60, 90, and 120 minutes. In all groups, a uniphasic enhancement pattern was observed in the outer and inner medulla of the left kidney, accompanied by a decrease of the area under the curve. CONCLUSION: Our results suggest that MR diffusion-weighted imaging and dynamic perfusion imaging are useful in identifying renal dysfunction following normothermic ischemia and reperfusion injury. PMID- 12766900 TI - Accuracy of cartilage and subchondral bone spatial thickness distribution from MRI. AB - PURPOSE: To assess three-dimensional measurement accuracy of articular cartilage (AC) and subchondral bone (SB) thickness from MRI. MATERIALS AND METHODS: A computer program was used to calculate AC and SB thickness from MRI (three dimensional spoiled gradient echo (SPGR),.31-mm resolution, 1-mm slice thickness) of six adult femoral heads. Specimens were imaged in five anatomical planes ranging between +30 degrees to -30 degrees from neutral and cut into 2-mm thick sections along the five anatomical planes. Faxitron x-ray was used to produce microradiographic (.05-mm resolution) images of the sections. RESULTS: In-plane measurement accuracy was.165 +/-.108 mm for AC thickness and.387 +/-.174 mm for SB thickness. Taking into account chemical-shift misregistration in SB thickness, accuracy of measurements improved to.213 +/- 128 mm. Out-of-plane (three dimensional) thickness accuracy of the model, assessed by numerical simulation, was.015 mm. However, three-dimensional thickness errors in specimens were.319 +/ .256 mm for AC and.253 +/-.183 mm for SB thickness. CONCLUSION: Errors in three dimensional AC thickness were attributed to volume-averaging effects caused by oblique intersection of the image plane with the joint surface. Errors in three dimensional SB thickness were attributed to chemical-shift artifact. We conclude that accuracy of AC thickness is within clinically acceptable standards but that more sophisticated pulse sequences are needed to improve the measurement of SB thickness. PMID- 12766899 TI - Contrast-enhanced MR angiography of the run-off vasculature: intraindividual comparison of gadobenate dimeglumine with gadopentetate dimeglumine. AB - PURPOSE: To compare intraindividually gadobenate dimeglumine (Gd-BOPTA) with gadopentetate dimeglumine (Gd-DTPA) for multi-station MR Angiography of the run off vessels. MATERIALS AND METHODS: Twenty-one randomized healthy volunteers received either Gd-BOPTA or Gd-DTPA as a first injection and then the other agent as a second injection after a minimum interval of 6 days. Each agent was administered at a dose of 0.1 mmol/kg bodyweight followed by a 25-mL saline flush at a single constant flow rate of 0.8 mL/second. Images were acquired sequentially at the level of the pelvis, thigh, and calf using a fast three dimensional (3D) gradient echo sequence. Source, subtracted source, maximum intensity projection (MIP), and subtracted MIP image sets from each examination were evaluated quantitatively and qualitatively on a segmental basis involving nine vascular segments. RESULTS: Significantly (P < 0.05) higher signal-to-noise and contrast-to-noise ratios were noted for Gd-BOPTA compared to Gd-DTPA, with the more pronounced differences evident in the more distal vessels. Qualitative assessmentrevealed no differences in the abdominal vasculature, a preference for Gd-BOPTA in the pelvic vasculature, and markedly better performance for Gd-BOPTA in the femoral and tibial vasculature. Summation of individual diagnostic quality scores for each segment revealed a significantly (P = 0.0001) better performance for Gd-BOPTA compared to Gd-DTPA. CONCLUSION: Greater vascular enhancement of the run-off vasculature is obtained after Gd-BOPTA, particularly in the smaller more distal vessels. Enhancement differences are not merely dose dependent, but may be due to different vascular enhancement characteristics of the agents. PMID- 12766901 TI - Simple anatomical measurements do not correlate significantly to individual peripheral nerve stimulation thresholds as measured in MRI gradient coils. AB - PURPOSE: To examine peripheral nerve stimulation (PNS) thresholds for normal human subjects in magnetic resonance imaging (MRI) gradient coils, and determine if observed thresholds could be predicted based on gross physiologic measurements. MATERIALS AND METHODS: PNS thresholds for 21 healthy normal subjects were measured using a whole-body gradient coil. Subjects were exposed to a trapezoidal echo-planar imaging (EPI) gradient waveform and the total change in gradient strength (DeltaG) required to cause PNS as a function of the duration of the gradient switching time (tau) were measured. Correlation coefficients and corresponding P values were calculated for the PNS threshold measurements against simple physiologic measurements taken of the subjects, including weight, height, girth, and average body fat percentage, in order to determine if there were any easily observable dependencies. RESULTS: No convincing correlations between threshold parameters and gross physiologic measurements were observed. CONCLUSION: These results suggest it is unlikely that a simple physiologic measurement of subject anatomy can be used to guide the operation of MRI scanners in a subject-specific manner in order to increase gradient system performance while avoiding PNS. PMID- 12766902 TI - Magnetic resonance imaging and proton MR spectroscopy of the brain in a patient with carbohydrate-deficient glycoprotein syndrome type I. AB - This is a report on a case of carbohydrate-deficient glycoprotein syndrome (CDGS) with neurological deficits. Magnetic resonance (MR) images showed remarkable atrophy of the cerebellum and brainstem, and hypointensity was seen in the pallidum on diffusion-weighted images (DWI), suggesting deposits of metal substances. In the cerebellum, proton MR spectroscopy (MRS) showed decreased concentrations of N-acetylaspartate and a complex of glutamine and glutamate (Glx) while the concentration of myo-inositol was increased, indicating neuronal impairment and gliosis. In the parietal lobe, there was an increased concentration of Glx, possibly reflecting dysfunction caused by liver injury. PMID- 12766903 TI - Evaluation of sodium T1 relaxation times in human heart. AB - PURPOSE: To evaluate the sodium longitudinal relaxation (T(1)) characteristics for myocardium and blood in humans. MATERIALS AND METHODS: Eleven healthy volunteers were examined by using a (23)Na heart surface coil at a 1.5 T clinical scanner equipped with a broadband spectroscopy option. (23)Na MR measurements were performed by using a three-dimensional spoiled gradient echo sequence (in plane resolution, 3.5 mm x 7 mm; slice thickness, 24 mm; TE, 3.1 msec; bandwidth, 65 Hz/pixel; TR, 21 to 150 msec). RESULTS: Longitudinal T(1) relaxation time components were 31.6+/-7.0 msec and 31.1+/-7.5 msec for myocardium and blood, respectively. CONCLUSION: (23)Na T(1) relaxation times of myocardium and blood can be determined in humans. The results are in agreement with values obtained from animal studies. PMID- 12766904 TI - Three-dimensional T1rho-weighted MRI at 1.5 Tesla. AB - PURPOSE: To design and implement a magnetic resonance imaging (MRI) pulse sequence capable of performing three-dimensional T(1rho)-weighted MRI on a 1.5-T clinical scanner, and determine the optimal sequence parameters, both theoretically and experimentally, so that the energy deposition by the radiofrequency pulses in the sequence, measured as the specific absorption rate (SAR), does not exceed safety guidelines for imaging human subjects. MATERIALS AND METHODS: A three-pulse cluster was pre-encoded to a three-dimensional gradient-echo imaging sequence to create a three-dimensional, T(1rho)-weighted MRI pulse sequence. Imaging experiments were performed on a GE clinical scanner with a custom-built knee-coil. We validated the performance of this sequence by imaging articular cartilage of a bovine patella and comparing T(1rho) values measured by this sequence to those obtained with a previously tested two dimensional imaging sequence. Using a previously developed model for SAR calculation, the imaging parameters were adjusted such that the energy deposition by the radiofrequency pulses in the sequence did not exceed safety guidelines for imaging human subjects. The actual temperature increase due to the sequence was measured in a phantom by a MRI-based temperature mapping technique. Following these experiments, the performance of this sequence was demonstrated in vivo by obtaining T(1rho)-weighted images of the knee joint of a healthy individual. RESULTS: Calculated T(1rho) of articular cartilage in the specimen was similar for both and three-dimensional and two-dimensional methods (84 +/- 2 msec and 80 +/- 3 msec, respectively). The temperature increase in the phantom resulting from the sequence was 0.015 degrees C, which is well below the established safety guidelines. Images of the human knee joint in vivo demonstrate a clear delineation of cartilage from surrounding tissues. CONCLUSION: We developed and implemented a three-dimensional T(1rho)-weighted pulse sequence on a 1.5-T clinical scanner. PMID- 12766905 TI - Mutation analysis of CDP, TP53, and KRAS in uterine leiomyomas. AB - Leiomyomas are the most common gynecologic tumors in women, but very little is known about their molecular pathology. We used single-stranded conformational polymorphism/heteroduplex analysis to analyze 42 unselected uterine leiomyomas for somatic mutations in all coding exons of the gene encoding CCAAT displacement protein (CDP), as well as exons 5-8 of TP53 and codons 1-36 and 38-80 of KRAS. No somatic mutations were identified in either TP53 or KRAS, indicating that disregulation of these genes is not required for leiomyomas development. Aberrant band shifts were identified in CDP, but these were all germline nonpathogenic variants that have been reported previously. There is good functional and genetic evidence indicating that CDP is a leiomyoma suppressor, but our data suggested that somatic mutations in this gene were rare in unselected uterine leiomyomas. It is possible that CDP belongs to a class of tumor suppressor in which loss of only one copy of the gene, either by genetic or epigenetic mechanisms, is sufficient to allow tumor growth. PMID- 12766906 TI - Identification of distinct and common gene expression changes after oxidative stress and gamma and ultraviolet radiation. AB - The human genome is exposed to many different kinds of DNA-damaging agents. While most damage is detected and repaired through complex damage recognition and repair machineries, some damage has the potential to escape these mechanisms. Unrepaired DNA damage can give rise to alterations and mutations in the genome in an individual cell, which can result in malignant transformation, especially when critical genes are deregulated. In this study, we investigated gene expression changes in response to oxidative stress, gamma (gamma) radiation, and ultraviolet (UV) radiation and their potential implications in cancer development. Doses were selected for each of the three treatments, based on their ability to cause a similar G(1) checkpoint induction and slow down in early S-phase progression, as reflected by a comparable reduction in cyclin E-associated kinase activity of at least 75% in logarithmically growing human dermal diploid fibroblasts. To investigate gene expression changes, logarithmically growing dermal diploid fibroblasts were exposed to either gamma radiation (5 Gy), oxidative stress (75 microM of tert-butyl hydroperoxide (t-butyl-OOH)), or UV radiation (UVC) (7.5 J/m(2)) and RNA was harvested 6 h after treatment. Gene expression was analyzed using the NIEHS Human ToxChip 2.0 with approximately 1901 cDNA clones representing known genes and expressed sequence tags (ESTs). We were able to identify common and distinct responses in dermal diploid fibroblasts to the three different stimuli used. Within our analysis, gene expression profiles in response to gamma radiation and oxidative stress appeared to be more similar than profiles expressed after UV radiation. Interestingly, equivalent cyclin E-associated kinase activity reduction with all the three treatments was associated with greater transcriptional changes after UV radiation than after gamma radiation and oxidative stress. While samples treated with UV radiation displayed modulations of their mitogen activated protein kinase (MAPK) pathway, gamma radiation had its major influence on cell-cycle progression in S-phase and mitosis. In addition, cell cultures from different individuals displayed significant differences in their gene expression responses to DNA damage. PMID- 12766907 TI - Modulation of the angiogenesis response through Ha-ras control, placenta growth factor, and angiopoietin expression in mouse skin carcinogenesis. AB - Tumor angiogenesis is governed by a complex balance of positive and negative angiogenic factors. Development of chemically-induced mouse skin tumors appears to be highly dependent on an early burst of neovascularization. We have previously shown that Ha-ras-driven vascular endothelial growth factor (VEGF) expression plays a pivotal role in this process. However, the status of other critical positive and negative angiogenic factors throughout skin tumorigenesis has not been studied to the same extent. In the present study, we show that another VEGF family member, placenta growth factor (PlGF), was highly upregulated at all tumor stages in a ras-dependent manner. The study of angiopoietin-1 (Ang 1) and angiopoietin-2 (Ang-2), ligands of receptor tyrosine kinase 2 (Tie-2), showed that while stroma-derived Ang-2 was increased, epidermal Ang-1 expression was completely abolished at early papilloma formation. Studies using epidermal tumor cell lines suggest that the disappearance of Ang-1 also depends on ras activation, extending the plethora of events controlled by this oncogene in mouse skin carcinogenesis. Our results indicated that tumor development occurred in a strong angiogenesis-prone scenario in which PlGF and Ang-2 acted cooperatively with VEGF, whereas the negative or stabilizing effect of Ang-1 was abrogated. A time-course sequence of expression of angiogenic factors expressed throughout tumor growth, as well as the identification of key signaling molecules triggering the angiogenic response, may contribute to the development and testing of antiangiogenic therapeutic strategies with this in vivo tumor model. PMID- 12766908 TI - Inactivation of helicase-like transcription factor by promoter hypermethylation in human gastric cancer. AB - Helicase-like transcription factor (HLTF), a member of the SWI/SNF (mating type switching/sucrose nonfermenting) chromatin-remodeling complex, is recently found to be inactivated by promoter hypermethylation in human colorectal cancer. However, the role of this putative tumor suppressor gene in other tumors has not been determined. We evaluated the role of HLTF promoter hypermethylation in gastric cancer. Expression of HLTF was examined by reverse-transcription (RT) polymerase chain reaction (PCR), and promoter hypermethylation in HLTF was determined by methylation-specific PCR. Bisulfite DNA sequencing was performed to determine the detailed methylation profiles of the promoter region. HLTF expression was lost in two of five gastric cell lines and in 13 (28%) of 46 primary gastric cancers. Accordingly, promoter hypermethylation was detected in the two cell lines and in nine of 13 gastric cancer samples. Of the ten normal gastric specimens and ten paired adjacent nonneoplastic tissues, methylation was detected in only one adjacent nonneoplastic tissue. Bisulfite DNA sequencing of the promoter region of HLTF showed that the CpG island was densely methylated in cell lines and cancer samples; this also appeared to correlate with expression level. Treatment of gastric cell lines that lacked HLTF expression with the demethylating agent 5-azacytidine (5-azaDC) restored HLTF expression. These results suggest that HLTF promoter hypermethylation is frequently demonstrated in human gastric cancer, and inactivation of HLTF or the chromatin-remodeling complex may play a crucial role in gastric carcinogenesis. PMID- 12766909 TI - Regulation of cell morphology and adhesion by the tuberous sclerosis complex (TSC1/2) gene products in human kidney epithelial cells through increased E cadherin/beta-catenin activity. AB - We investigated the effects of overexpression of the tuberous sclerosis-1 and -2 (TSC1/2) gene products (hamartin and tuberin, respectively) in the human kidney epithelial cell line 293 with an inducible expression system. As we had observed previously in fibroblasts, 293 cells overexpressing hamartin and/or tuberin grew more slowly in vitro. However, here we also observed that the 293 overexpressing cells underwent a dramatic morphological change in which groups of cells formed compact clusters. The overexpressing cells also displayed decreased dissociation and increased reaggregation in vitro. These changes were found to be associated with an increased level of E-cadherin, which is known to regulate cell-cell interactions in epithelial cells, and of its binding partner beta-catenin. Consistent with the role of E-cadherin in these effects, we found that the observed changes in 293 cell morphology, dissociation, and adhesion were calcium dependent, and were reproduced by overexpression of E-cadherin. In contrast, overexpression of TSC1 in rat embryo fibroblasts, which lack E-cadherin, failed to elicit the same changes as in 293 cells. We conclude that the hamartin/tuberin complex exerted a direct effect on the morphology and adhesive properties of 293 cells through regulation of the level and/or activity of cellular E-cadherin/beta catenin. PMID- 12766910 TI - Formation of an active form of the interleukin-2/15 receptor beta-chain by insertion of the intracisternal A particle in a radiation-induced mouse thymic lymphoma and its role in tumorigenesis. AB - Although many reports suggest that aberrant regulation of cytokine signaling pathways via the interleukin-2 receptor (IL-2R) induces tumorigenic transformation, constitutively active IL-2R in tumors has not been reported. We searched for genomic alteration of the IL-2/15R beta-subunit gene (IL-2/15R beta) in cytokine-independent cell lines established from radiation-induced mouse thymic lymphomas. In the TL34 cell line and its primary tumor, one of the IL 2/15R beta alleles was rearranged by the insertion of an intracisternal A particle (IAP) retrotransposon. The IAP-IL2/15R beta chimeric gene expressed chimeric mRNA in which IAP-coding Gag-Pol mRNA was fused to IL-2/15R beta mRNA and coded for Gag-Pol-IL-2/15R beta chimeric protein. Forced expression of the Gag-Pol-IL-2/15R beta chimeric cDNA in a mouse cytotoxic T-cell line (CTLL-2) converted IL-2-dependent cell growth to IL-2-independent growth, suggesting that the chimeric protein activates some of the IL-2 signaling pathways necessary for cell proliferation. Downregulation of the expression of the Gag-Pol-IL-2/15R beta chimeric protein in TL34 by antisense RNA inhibited cell growth, and concomitantly reduced the level of c-myc protein. These results suggest that the Gag-Pol-IL-2/15R beta is a constitutively active form that transmits proliferative signals by expressing downstream target genes, including c-myc. Thus, we demonstrated that the chimeric receptor gene produced by the insertion of an IAP functions as an oncogene by providing IL-2-independent autonomous growth potential. PMID- 12766911 TI - The identification of trends in the utilisation of mental health services by elderly: a Dutch case register study. AB - OBJECTIVE: In view of the rapid ageing of the population any changes in the use of mental health services by the elderly became increasingly important for policy development. This study aimed at the supply of information about trends in the numbers of elderly clients, the services they used and the volume and pattern of service utilisation. METHODS: Details of elderly users and their use of community and hospital-based services between 1990 and 1999 were retrieved from the Groningen case register. Developments in population size and age distribution in the register area were taken into account, as were the unit costs of mental health services. RESULTS: Large age specific changes were found that caused only the expenditures on the oldest elderly to increase due to a shift from outpatient clinics to prolonged psychogeriatric day treatment and inpatient care. Comparatively young elderly used fewer inpatient services and more community care. The number of new elderly clients declined progressively. In some age groups treated prevalence also decreased, but to a lesser extent, because of a prolonged use of mental health services. CONCLUSIONS: Study results seemed well in accordance with mental health policy as to deinstitutionalization and active ageing. Research on the effect of mental health care on life expectancy and the time lag between the intake of mental health providers and treated prevalence was proposed in order to improve the prediction of future service use by elderly. PMID- 12766912 TI - Validation of the modified telephone interview for cognitive status (TICS-m) in Hebrew. AB - INTRODUCTION: The validity of the Hebrew version of the Telephone Interview for Cognitive Status-Modified (TICS-m) for Mild Cognitive Impairment (MCI), for dementia, and for cognitive impairment (either MCI or dementia) was investigated. METHODS: Of the 10 059 who took part of the Israel Ischemic Heart Disease Cohort, 1902 of the 2901 survivors in 1999 had TICS-m interviews. Those with a score of 27 or below and a random sample with a score of 28 or 29 were invited to have a physician's examination for the diagnosis of dementia. The analysis was performed on the 576 who agreed. RESULTS: Based on physician's diagnosis, 269 were diagnosed as suffering from dementia, 128 as suffering from MCI, and 179 were diagnosed with no cognitive impairment. The TICS-m Hebrew version's internal consistency was very high (Cronbach's alpha = 0.98) and showed a strong convergent validity with the MMSE (r = 0.82; p < 0.0005). The sensitivity was 100% for each of the conditions. Finally, after controlling for age, education and hearing impairment, TICS-m was a strong predictor of dementia, MCI and cognitive impairment. CONCLUSION: At a cut-off of 27/50 the Hebrew version of the TICS-m is a useful screening instrument to identify subjects suffering from mild cognitive impairment, dementia and cognitive impairment (MCI or dementia). PMID- 12766913 TI - Visuospatial impairment in dementia with Lewy bodies and Alzheimer's disease: a process analysis approach. AB - BACKGROUND: Reports of differential impairments on visual-construction tasks in dementia with Lewy bodies (DLB) and Alzheimer's disease (AD) are sometimes controversial, whereas visual-perceptual data are lacking. The existence of different clinical sub-groups of DLB has been hypothesized to explain the discrepancies among the cognitive results. The goal of this study was to compare the visual-perceptual performance of subjects with DLB with predominant psychosis, DLB with predominant parkinsonian features and AD. METHODS: This is a cross-sectional neuropsychological study with between diagnostic group comparisons. The Benton Judgement Line Orientation (BJLO) test was administered to four DLB patients with predominant psychosis (DLB-psy), four DLB subjects with predominant parkinsonian features (DLB-PD), and 13 patients with AD. An analysis of error types was applied to the results of the BJLO with QO1, QO2, QO3, QO4 (visual attention) errors, as well as VH, IQO, IQOV, and IQOH (visual-spatial perception) errors. RESULTS: A MANOVA showed significant differences between the DLB, and AD groups on the number of VH (F = 6.049, df = 1,19, p = 0.024), IQOH (F = 4.645, df = 1,19, p = 0.044) and QO1 (F = 4.491, df = 1,19, p = 0.047) errors, but no difference on the total score of the BJLO. Another MANOVA and post hoc Student-Newman-Keuls analyses demonstrated that the DLB-psy sub-group made significantly more VH and IQOH errors than AD and the DLB-PD subjects. CONCLUSIONS: Subjects with DLB and psychosis have more severe visual-perception (VH errors) impairments than subjects with DLB and predominant parkinsonian features, and AD subjects. PMID- 12766915 TI - The early stage of vascular dementia: significance of a complete therapeutic program. AB - THEORY: Mild manifestations of vascular dementia are relatively widespread among the old-age population. Drug therapy for this disorder is insufficiently effective at present, making it necessary to improve the treatment of such patients by means of an adequate psychotherapeutic and rehabilitation program. AIMS: To assess the role of psychotherapeutic and rehabilitation measures in improving outcome in the early stage of vascular dementia. METHODS: The test and control cohorts (each n = 95) were matched for characteristics found to be significant for outcome of treatment (the relevant predictors were found using Multiple Regression Analysis). The cohorts differed in the amount of the psychotherapy and rehabilitation received. To assess the effects of treatment the SCAG scale and a special scale for evaluating the level of activity in the household were used. RESULTS: In the test cohort the patients' condition improved significantly compared with the controls. Even in case of only slight therapeutic reduction of psychopathologic symptoms the systematic use of psychotherapeutic and rehabilitation methods favoured preserving a certain level of activity in the household and for some patients even improving it. CONCLUSIONS: The results of therapy at the early stage of vascular dementia to a great extent depend on carrying out a full-scale psychotherapeutic and rehabilitation program. The positive significance of the latter is connected not only with improvement of clinical indices but also with direct optimizing the every day life activity level. PMID- 12766914 TI - Service provision for elderly depressed persons and political and professional awareness for this subject: a comparison of six European countries. AB - OBJECTIVE: Under-treatment of depression in late-life is a subject of rising public health concern throughout Europe. This study investigates and compares the availability of services for depressed elderly persons in Denmark, France, Germany, Sweden, Switzerland and the UK. Additionally, it explores factors that might contribute to an adequate services supply for depressed elderly people. METHOD: Review of the literature and guide supported expert interviews. Analysis of the practice of care provision for depressed elderly persons and of indicators for political and professional awareness, such as university chairs, certification processes and political programmes in gerontopsychiatry. RESULTS: Only Switzerland and the UK offer countrywide community-oriented services for depressed elderly persons. Clinical experience in treating depression in late life is not regularly acquired in the vocational training of the concerned professionals. Indicators suggest that the 'medical society' and health politics in Switzerland and the UK regard psychiatric disease in the elderly more importantly than it is the case in the other investigated countries. CONCLUSIONS: Service provision for depressed elderly persons seems to be more elaborated and better available in countries where gerontopsychiatry is institutionalised to a greater extend in the 'medical society' and health politics. PMID- 12766916 TI - CSF phosphorylated tau--does it constitute an accurate biological test for Alzheimer's disease? AB - INTRODUCTION: There is considerable interest in developing a diagnostic test which could differentiate between early Alzheimer's disease (AD) and other causes of memory impairment with more than 80% sensitivity and 80% specificity. OBJECTIVE: To review the studies that have examined CSF phosphorylated tau as diagnostic test of AD vs clinically representative comparison groups. METHOD: A critical review of the literature using Embase, Web of Science, Medline and Psychinfo databases supplemented by handsearching and contact with experts in the field. RESULTS: CSF phosphorylated tau is a marker of AD that improves upon the utility of CSF total tau and clinical examination alone. Studies have found high levels of tau phosphorylated at Threonine 231 and/or Serine 199 in AD but not in other causes of dementia, in depression or in healthy elderly controls. Of particular interest, the test appears equally valid in cases of early AD as in moderate or late stages and may also be of use in predicting future decline in subjects with mild cognitive impairment. CONCLUSION: CSF phosphorylated tau is a promising diagnostic test for AD but this requires replication using pathologically confirmed cases. PMID- 12766917 TI - Prevalence of sub-threshold depression in elderly patients with chronic obstructive pulmonary disease. AB - OBJECTIVES: We hypothesized that COPD patients with sub-threshold depression would have levels of disability and impaired quality of life approaching that for major depression and significantly greater than for non-depressed COPD patients. SETTING: A university teaching hospital METHOD: 137 outpatients (69 men), with a mean age of 73 years (range 60-89 years) with symptomatic irreversible, moderate to severe COPD were recruited. Subjects were interviewed using the Geriatric Mental State Schedule (GMS), a structured psychiatric interview schedule, along with its diagnostic algorithm AGECAT. A GMS/AGECAT score of 3 or more is indicative of a case-level of depression, a GMS/AGECAT score of 1-2 indicates sub threshold depression and GMS/AGECAT of 0, no depression. Physical disability was measured by the Manchester Respiratory Activities of Daily Living questionnaire (MRADL) and quality of life was assessed by the Breathing Problems Questionnaire (BPQ). RESULTS: Mean (SD) one second forced expiratory volume was 0.89 (0.33) litres. The prevalence of GMS/AGECAT case-level depression (>or= 3) was 57 cases (42%); of GMS/AGECAT sub-threshold depression (1-2) 34 (25%); and GMS/AGECAT non depression (0) 46 (33%). Comparison of MRADL score in the three groups (mean, 95% confidence intervals) revealed [GMS >or= 3 = 9.9 (8.4 to 11.3) vs GMS = 1-2, 12.9 (11.2 to 14.4) vs GMS = 0, 15.6 (14 to 16.6) p < 0.0001]. BPQ scores (mean, 95% confidence intervals) showed [GMS >or= 3 = 54 (50 to 57) vs GMS = 1-2, 40 (36.3 to 44) GMS = 0, 33 (30.6 to 36.7) p < 0.0001]. There was no significant difference in FEV(1) between the three groups. CONCLUSION: Sub-threshold depression accounted for 25% of the sample. In this study disability associated with sub-threshold depression in patients with COPD was intermediate to that associated with case-level depression and no with depression and significantly worse than in the latter group. Sub-threshold depression is associated with substantial morbidity in COPD. PMID- 12766918 TI - Phantom boarder symptom in dementia. AB - OBJECTIVE: Phantom boarder symptom (PBS), which has been labelled a misidentification and is commonly observed in cases of Alzheimer's disease, is the perceptual dysfunction where it is imagined that there are people in the home. It may also be encountered in cases of vascular dementia and other variants of this cognitive dysfunction. This study was undertaken to investigate PBS frequency and characteristics for a sample of dementia inpatients, and to determine the strength of the association with other psychotic symptoms. METHODS: The sample population consisted of 240 dementia patients admitted to our geropsychiatric ward. Patients with and without PBS were compared in terms of general characteristics and psychotic symptoms. RESULTS: Of the 240 dementia patients, PBS was identified in 56 (23.3%). There were no significant gender differences between the PBS and non-PBS groups. Means for age and onset-age were higher for the PBS group, and the prevalence of hallucinations and other misidentifications was greater compared to the non-PBS analog. Occurrence of PBS was not significantly associated with delusions or physical aggression. CONCLUSIONS: PBS is a common symptom in dementia of various etiologies. Our association findings suggest that it may be more productive to classify PBS as a type of misidentification, instead of as a delusion. PMID- 12766919 TI - Antidepressant-associated mania in late life. AB - BACKGROUND: Elderly patients can present with mania for the first time late in life, and some elders treated with antidepressants can present with mania. Clinical characteristics of antidepressant-associated mania (AAM) in late life have not been examined. OBJECTIVES: The aims of the study were to identify elders with AAM and to compare selected clinical characteristics to those of manic elders who had not been treated with an antidepressant. We hypothesized that AAM patients would have later age at presentation of bipolar disorder. METHODS: We retrospectively reviewed inpatients with manic disorder who were aged >or=60 years. The sample was selected from admissions prior to 1990. RESULTS: AAM patients (n = 11) were more often experiencing first manic episode, and they had later age at onset of first manic episode, compared to non-AAM patients (n = 46). Most of the AAM patients had been treated with tricyclic agents. CONCLUSIONS: These preliminary findings invite further investigation. Related studies may contribute to risk-benefit analyses for the use of particular antidepressants in the elderly. Also, first episode mania in late life may prove to be a useful model of vulnerability to AAM. PMID- 12766920 TI - Does aspirin affect outcome in vascular dementia? A retrospective case-notes analysis. AB - BACKGROUND: Ischaemic vascular dementia shares risk factors with stroke. There is evidence that control of these risk factors may prevent or alter the course of vascular dementia. OBJECTIVE: To assess the effect of regular low-dose aspirin on outcomes for patients with vascular dementia. DESIGN: Retrospective analysis of hospital case-notes with further outcome information from telephone calls to general practitioners, social services and institutions. Comparison of outcomes for aspirin-treated and untreated patients. SETTING: One North London NHS Trust. PATIENTS: Seventy-eight patients with clinician's diagnosis of ischaemic vascular dementia, discharged from acute inpatient units between 1 January 1995 and 31 December 1997; 38 on aspirin. MAIN OUTCOME MEASURES: Survival times from dementia onset to institutionalization and death. RESULTS: Median survival time to institutionalization was 28 months and to death was 52 months. There was no overall difference between aspirin and non-aspirin groups for these outcomes. When data were stratified for social status, i.e. living alone or with carer when last at home, differences emerged for those living with carer. Aspirin was associated with a trend towards increased time to institutionalization (39 vs 22 months, p < 0.09) and a significant advantage in time to death (71 vs 27 months, p = 0.02). These effects were non-significant after statistical adjustment for confounding variables. CONCLUSIONS: The results support but do not prove a role for regular, low-dose aspirin in improving both life expectancy and survival at home for patients with vascular dementia. Compliance may be better in those living with a carer. Larger, prospective studies should be performed to confirm these findings. Cognitive and behavioural outcomes should also be studied. PMID- 12766922 TI - The effect of Qigong on general and psychosocial health of elderly with chronic physical illnesses: a randomized clinical trial. AB - OBJECTIVES: Based on the model by Tsang et al. (2002) which summarized the etiological factors and consequences of depression in elderly with chronic physical illnesses, a randomized clinical trial of a special form of Qigong (The Eight Section Brocades) was conducted to assess if it improved the biopsychosocial health of participants. DESIGN: 50 geriatric patients in sub acute stage of chronic physical illnesses were recruited and randomly assigned into the intervention and control group. The intervention group was given a 12 week period of Qigong practice while the control group was given traditional remedial rehabilitation activities. RESULTS: The intervention group participants expressed improvement in physical health, ADL, psychological health, social relationship, and health in general as reflected by scores of the Perceived Benefit Questionnaire and informal feedback. CONCLUSION: Although results are not significant in the generalization measures, it may be due to small effect size, small sample size, and short intervention period. Although not all of the hypotheses are supported, this report shows that Qigong (the Eight Section Brocades) is promising as an alternative intervention for elderly with chronic physical illness to improve their biopsychosocial health. More systematic evaluation with larger sample size and longer period of intervention is now underway in Hong Kong. Results will be reported once available. PMID- 12766921 TI - The impact upon extra-pyramidal side effects, clinical symptoms and quality of life of a switch from conventional to atypical antipsychotics (risperidone or olanzapine) in elderly patients with schizophrenia. AB - BACKGROUND: Atypical antipsychotics are commonly used in the management of schizophrenia in late life with evidence suggesting they induce lower rates of motor disturbance, but have similar efficacy to conventional antipsychotics. Trials in the elderly have been either retrospective, small, of short duration or of a single-arm design. AIMS: To demonstrate the effects upon motor side-effects, efficacy, safety and quality of life (QOL) of switching elderly patients with schizophrenia from conventional antipsychotics to olanzapine or risperidone. METHODS: Elderly patients with schizophrenia were randomly allocated to olanzapine or risperidone and followed through an open-label crossover period. Between and within group intention to treat analyses were conducted. RESULTS: 66 patients were randomised (mean age 69.6 [SD +/- 6.2]). Four (11.8%) patients on olanzapine and 8 (26.7%) patients on risperidone failed to complete the crossover because of treatment failure [Odds Ratio (OR) = 2.73[0.73-10.2] p = 0.14]. The mean doses upon completion of switching in each arm were 9.9 mg (SD = 4.2) and 1.7 mg (SD = 1.2) for olanzapine and risperidone respectively. In both arms there was improvement in Parkinsonism, though only olanzapine was associated with a reduction in dyskinetic symptoms. The Brief Psychiatric Rating Scale, Scale for the assessment of Negative Symptoms and Montgomery and Asberg Depression Rating Scale scores all improved through the crossover period in both arms with no between group differences. Treatment with olanzapine was associated with a better response over risperidone on the psychological domain of the World Health Organisation-Quality Of Life [Brief] (WHO-QOL-BREF) scale ( p = 0.02). Patients in the olanzapine arm also demonstrated improvement from baseline in the WHO-QOL BREF physical, psychological and health satisfaction domains, but risperidone had no effect on any Quality of Life (QOL) measure. CONCLUSIONS: After switching from a conventional antipsychotic, olanzapine and risperidone were associated with improvement in core symptoms of schizophrenia and motor side effects. Subjects switched to olanzapine were more likely to complete the switching process and show an improvement in psychological QOL. PMID- 12766923 TI - A population based study on the intra and inter-rater reliability of the clock drawing test in Brazil: the Bambui Health and Ageing Study. AB - BACKGROUND: Reliability should be considered when selecting a scoring system since it influences validity. CDT reliability has rarely been assessed in population based studies and in developing countries. The aim of the present study was to determine intra and inter-rater reliabilities of the CDT scored by the Shulman (2000) method, in elderly with very low formal educational level from Brazil. METHODS: CDTs performed by a random sample of 202 subjects of a population-based cohort of elderly were scored on two occasions by the same rater and by two independent raters. Reliability was measured using the kappa statistic, weighted kappa and the intraclass correlation coefficient. Data were stratified according to gender, age and schooling level. RESULTS: Intra and inter rater reliabilities were excellent when CDTs were classified as 'normal' (scores 4 or 5) or 'abnormal' (scores 0 to 3) (kappa = 0.99 and 0.94, respectively) and were in the good to excellent range when scored from 0 to 5 (kappa = 0.88 and 0.74, respectively). Difficulties in distinguishing between scores 4 and 5, and a low proportion of score 1 tests were found. CONCLUSIONS: The CDT scored by the Shulman (2000) method appears to have good to excellent reliability in an elderly population with very low formal educational level. However, difficulties in distinguishing between scores 4 and 5, and a low proportion of score 1 tests suggest these scores may not be totally adequate for this population. Further studies are necessary to determine the consistency of our results in similar populations. PMID- 12766924 TI - Lowest serum cholesterol values are associated with depressive symptoms but not with mood disorders. PMID- 12766925 TI - Cholinesterase inhibitors in the treatment of dementia. PMID- 12766926 TI - Re: Bell-McGinty et al standard measures of executive function in predicting instrumental activities of daily living in older adults. PMID- 12766928 TI - Animal models of depression: are there any? AB - Simple tests for antidepressant-like activity, such as 5-HTP-induced syndrome or reserpine-induced hypomotility, are often mechanism-based tests, pharmacologically specific for certain known classes of therapeutically successful antidepressant agents. Many of these behavioural assays have been superseded by neurochemical techniques such as in vivo microdialysis. In contrast to these mechanistic-based models, investigators have also endeavoured to reproduce in the laboratory, factors that are believed to precipitate depression in people. It is a strong assumption in this approach that depression is a response to stress. This strategy profiles the consequences of chronic stress particularly psychosocial stress or early life events, in order to reproduce in animals the behavioural signs and pathologies associated with depression. The advances in the social psychological, clinical pathological and new areas such as neuroimaging research offer the possibility of establishing more sophisticated models for depression in animals with a broader range of biomarkers from the immunological and endocrinological to neurochemical and behavioural. Combining these novel insights with more traditional tests of depression may not only increase our understanding of the neurobiology of depression but also afford more precise and predictive preclinical models of depression. The responsiveness of different strains or genetically modified animals to stress is likely to be a key area of study. Furthermore we must look to individual differences in subjects, even within the same strain, to more fully understand why some individuals show pathological responses to stress whereas others appear unaffected. Conversely in validating our models using currently available treatments we must include the concept of non-responders so as not to disregard models that may extend therapeutic possibilities in these patients. PMID- 12766929 TI - Milnacipran plasma levels and antidepressant response in Japanese major depressive patients. AB - The relationship between antidepressant effects and plasma levels of milnacipran was studied in 49 cases of major depression without psychotic features during 6 weeks of milnacipran treatment. The daily dose of milnacipran was 50 mg/day for the first week, and up to 100 mg/day thereafter. Depressive symptoms were evaluated by the Montgomery and Asberg depression rating scale (MADRS) before treatment and at 1, 2, 4 and 6 weeks after the beginning of this study. Thirty four patients (69.4%) were responders (defined as a 50% or greater decrease in the baseline MADRS score). Significant differences of MADRS scores were seen from 1 week after the beginning of this study (p=0.004, unpaired t-test) between responders and nonresponders. The mean plasma milnacipran level of responders, 82.0 +/- 29.4 ng/ml, was similar to that of non-responders, 78.6+/-23.1 ng/ml; there was no significant difference between responders and nonresponders. Neither a significant linear nor a curvilinear relationship was obtained between the final MADRS score and the plasma levels of milnacipran. Although there was no significant relationship between the plasma levels of milnacipran and the antidepressant response, milnacipran should be considered an efficacious agent in the treatment of major depressive patients. PMID- 12766930 TI - A placebo controlled investigation into the effects of paroxetine and mirtazapine on measures related to car driving performance. AB - OBJECTIVE: To assess the effects of paroxetine and mirtazapine on psychometric performance related to car driving, including an on-the-road test of BRT. METHOD: In a 4-way, double blind randomised crossover study, 12 healthy volunteers received paroxetine 20mg mane, mirtazapine 15mg/30mg nocte (comparator), mirtazapine 15mg mane/15 mg b.i.d.(verum) and placebo over a 5 day period with a washout period of 7 days between treatments. Psychometric assessments included 'on-the-road' BRT (BRT), CFF (CFF), CRT (CRT) and subjective measures of sedation and sleep parameters. RESULTS: Paroxetine had no significant effect on BRT compared with placebo. Although subjective ratings of sleep quality and sedation were impaired, there were significant improvements in both CFF and the recognition reaction component of CRT with paroxetine. Mirtazapine 15mg/30mg nocte impaired laboratory performance and some subjective tests. Mirtazapine 15mg mane/15mg b.i.d. improved sleep, but significantly impaired all other measures. CONCLUSION: Paroxetine 20 mg/day has no psychomotor or behavioural toxicity and has no negative impact on BRT. Further research into the chronic and sub-chronic effects of mirtazapine is needed to establish the clinical significance of these results. PMID- 12766931 TI - Efficacy of nefazodone in the treatment of neuroleptic induced extrapyramidal side effects: a double-blind randomised parallel group placebo-controlled trial. AB - Many atypical antipsychotics show antagonism at both serotonergic and dopaminergic neurones and show fewer extrapyramidal side effects (EPS). Nefazodone blocks postsynaptic 5HT2A receptors and weakly inhibits serotonin reuptake. This study aimed to elucidate the role of nefazodone in the treatment of antipsychotic-induced EPS. The trial was a double-blind, randomised, placebo controlled trial of patients requiring antipsychotic treatment with haloperidol 10mg daily; from which a subgroup of patients who developed EPS were selected for the study. Patients were randomised to add-on therapy with either placebo (n=24) or nefazodone (n=25) 100mg bd. EPS were measured on days 0, 3 and 7 using the Simpson Angus, Barnes akathisia, abnormal involuntary movement and Chouinard scales. Nefazodone significantly reduced EPS as measured by both the Simpson Angus scale and CGI (p=0.007 and 0.0247, respectively). Akathisia and tardive dyskinesia did not differ between the two groups (p=0.601; p=0.507, respectively). These results suggest the role of 5HT2 antagonism in the mechanism of action of atypical antipsychotics with respect to lowering rates of drug induced EPS. In addition, a therapeutic role for nefazodone is suggested in the treatment of antipsychotic-induced EPS. PMID- 12766932 TI - Switching to tianeptine in patients with antidepressant-induced sexual dysfunction. AB - Sexual side effects are frequent and are recently being considered as effects of antidepressant treatment. One method to improve the sexual dysfunction associated with the use of antidepressants is to change to another antidepressant. In the present work, the consequences of switching to tianeptine in patients with antidepressant-induced sexual dysfunction were studied. The study group comprised 23 patients with major depressive disorder who experienced antidepressant-induced sexual dysfunction. These antidepressants were stopped and switched to tianeptine (12.5mg x 3/day). All patients were screened by using the clinical global impression-improvement scale (CGI-I), the Hamilton depression rating scale (HAM D) and the Arizona sexual experience scale (ASEX) at the beginning of the study, and at weeks 4 and 8. No patient failed to tolerate 37.5mg of tianeptine or to complete the study except for one patient becoming pregnant. Paired t-tests revealed a significant difference between baseline and week 4 or week 8 in scores on both the HAM-D and ASEX. At 8 weeks, six patients were rated as very much improved (CGI-I=1) and ten patients were rated as much improved (CGI-I=2). Thus, with a CGI-I score of 2 or less used to indicate a positive response, 72.7% of the patients were responders. The results suggest that switching to tianeptine appears to be useful for alleviating sexual dysfunction caused by other antidepressants. PMID- 12766933 TI - Specific memory deficits in ecstasy users? The results of a meta-analysis. AB - A meta-analysis was carried out on the possible functional neurotoxic effects of ecstasy use in humans on verbal short-term memory (STM), verbal long-term memory (LTM), processing speed (RT) and % errors (attention). To that end studies were found on the effect of ecstasy that fulfilled the criteria for a meta-analysis (number of subjects, means and standard deviations of the dependent variables). Ten studies were included on STM, ten on LTM, eight on RT and eight on % errors (attention). In addition meta-regression analyses were carried out on the effect sizes with total lifetime ecstasy consumption (TLEC) as predictor. It was found that in all four meta-analyses the mean effect size (ES) was significant: ecstasy users had lower verbal STM and LTM scores, reacted slower and made more errors. The meta-regression coefficients were not significant, indicating no support for a linear relationship between the mean ES values and TLEC, leaving open the possibility for a stepwise relationship. Additional meta-analyses on ecstasy groups that did not differ in lifetime cannabis consumption showed that only the ES for LTM became insignificant. This suggests that ecstasy use does not decrease LTM, but the number of studies on which this conclusion is based was very low. PMID- 12766934 TI - Relationship of plasma amphetamine levels to physiological, subjective, cognitive and biochemical measures in healthy volunteers. AB - Acute administration of the stimulant dextro-amphetamine produces multiple physiological, subjective cognitive and biochemical changes. These effects are similar to those seen in mania, and may be a useful model for mania. The aim of the present study was more fully to determine the multiple effects of dextro amphetamine and to relate these to changes in plasma levels of the drug. In a double-blind, placebo-controlled crossover study in 25 healthy volunteers (ages 18-45), the effects of 25 mg of oral dextro-amphetamine were examined. Physiological, subjective, cognitive changes, concentrations of amino acids and metabolites of biogenic amines period were related to changes in plasma amphetamine concentrations over 500 min. Peak concentrations of dextro amphetamine occurred at 2.5-3.5 h post-administration and levels decreased to 75% of peak value after 500 min. The results from the present study indicate that the subjective psychological, cognitive and blood pressure changes frequently did not mirror the time course of plasma levels of the drug. Thus, there was no clear-cut relationship between plasma levels and effects. In addition, dextro-amphetamine caused no significant changes in amino acids or amino metabolite concentrations. In conclusion, while dextro-amphetamine administration definitely causes several changes which are seen in mania, there remain some physiological and metabolic differences between these two conditions. PMID- 12766935 TI - Olanzapine induced neuroleptic malignant syndrome--a case review. AB - Neuroleptic malignant syndrome is the rarest and the most serious of the neuroleptic induced movement disorders. Although potent neuroleptics are more frequently associated with NMS, atypical antipsychotic drugs may also be a cause of NMS. Three databases were searched using the terms 'olanzapine' and 'neuroleptic' 'malignant syndrome'. Case reports were selected and reviewed from among all articles that fulfilled the search criteria. Twenty six cases were reviewed. Twenty cases fulfilled the criteria published by Sachdev et al. Olanzapine was the most probable cause of NMS in 16 cases. The absence of rigidity was described in only two of 16 highly probable olanzapine induced NMS cases, which is not as often as it is reported in clozapine associated NMS (36%). It was found that prior NMS is an important risk factor in NMS. PMID- 12766936 TI - Treatment of cervical dystonia by olanzapine. PMID- 12766937 TI - Repetitive transcranial magnetic stimulation is effective following repeated courses in the treatment of major depressive disorder--a case report. AB - Repetitive transcranial magnetic stimulation (rTMS) is a relatively new treatment modality for psychiatric patients. rTMS was demonstrated to be effective in the treatment of depression. However, longitudinal outcome studies have not yet been published. Relapse rates are higher in depressed patients and most of them do not respond to the same treatment with similar success. In this report we present a patient, who experienced relapse with the various conventional drug treatments, but responded well to rTMS at three different points in time. PMID- 12766938 TI - Stereotypic and self-injurious behavior in rhesus macaques: a survey and retrospective analysis of environment and early experience. AB - Abnormal behavior in captive rhesus monkeys can range from active whole-body and self-directed stereotypies to self-injurious behavior (SIB). Although abnormal behaviors are common in singly-housed rhesus monkeys, the type and frequency of these behaviors are highly variable across individual animals, and the factors influencing them are equally varied. The purpose of this investigation was to survey abnormal behavior in a large population of rhesus macaques, to characterize the relationship between stereotypies and self-injury, and to identify potential risk factors for these aberrant behaviors. Behavioral assessments of 362 individually housed rhesus monkeys were collected at the New England Regional Primate Research Center (NERPRC) and combined with colony records. Of the 362 animals surveyed, 321 exhibited at least one abnormal behavior (mean: 2.3, range: 1-8). The most common behavior was pacing. Sex differences were apparent, with males showing more abnormal behavior than females. SIB was also associated with stereotypies. Animals with a veterinary record of self-injury exhibited a greater number of self-directed stereotypies than those that did not self-injure. Housing and protocol conditions, such as individual housing at an early age, longer time housed individually, greater number of blood draws, and nursery rearing, were shown to be risk factors for abnormal behavior. Thus, many factors may influence the development and maintenance of abnormal behavior in captive primates. Some of these factors are intrinsic to the individual (e.g., sex effects), whereas others are related to colony management practices, rearing conditions, and research protocols. PMID- 12766939 TI - Influence of estradiol on cortisol secretion in ovariectomized cynomolgus macaques (Macaca fascicularis). AB - In an investigation of cortisol secretion in fully mature, ovariectomized cynomolgus monkeys (Macaca fascicularis), we compared monkeys that were given either placebo (OVX, n = 26) or 17beta estradiol (E(2 )) (EST, n = 26) in a daily oral dose. Serum cortisol concentrations were measured prior to the experimental manipulation and 3, 6, 9, and 12 months following initiation of treatment. Pretreatment cortisol values did not differ between groups. Assessment of the treatment period values revealed that cortisol concentrations were significantly higher ( approximately 10%) in the EST than in the OVX monkeys. Cortisol also varied significantly across periods of sampling. This time-dependent variation was attributable to elevations in months 6 and 9 (when daylight was generally long), relative to months 3 and 12 (when daylight was relatively short). The modest stimulatory effect of estrogen on corticosteroid production observed in this study is consistent with what has been seen in women, and contrasts with the more robust effects observed in New World monkeys. The possible relationship between season and cortisol secretion observed here has not been previously described in monkeys. PMID- 12766940 TI - Four-year study of controlled timed breeding of rhesus monkeys (Macaca mulatta). AB - As part of the timed breeding colony at Tulane National Primate Research Center, exogenous progesterone administration (5 mg/day for 10 days) has been used to select conception dates by inducing artificial luteal phases in female rhesus monkeys. A retrospective analysis of data obtained during four breeding seasons (1998-2001) revealed that conceptions occurred an average of 18 days after the last administration of progesterone. The age of the female to be bred, previous pregnancy history, and timing of breeding during the breeding season were determined to be critical factors in the success of the procedure. The benefit of this method of timed breeding is that it does not require tracking of menstrual cycles, which can be labor-intensive and requires that animals be monitored several months in advance of breeding to determine each female's individual cycle length. It also provided an efficient use of breeding-age males. PMID- 12766941 TI - The Ciona intestinalis genome: when the constraints are off. AB - The recent genome sequencing of a non-vertebrate deuterostome, the ascidian tunicate Ciona intestinalis, makes a substantial contribution to the fields of evolutionary and developmental biology.1 Tunicates have some of the smallest bilaterian genomes, embryos with relatively few cells, fixed lineages and early determination of cell fates. Initial analyses of the C. intestinalis genome indicate that it has been evolving rapidly. Comparisons with other bilaterians show that C. intestinalis has lost a number of genes, and that many genes linked together in most other bilaterians have become uncoupled. In addition, a number of independent, lineage-specific gene duplications have been detected. These new results, although interesting in themselves, will take on a deeper significance once the genomes of additional invertebrate deuterostomes (e.g. echinoderms, hemichordates and amphioxus) have been sequenced. With such a broadened database, comparative genomics can begin to ask pointed questions about the relationship between the evolution of genomes and the evolution of body plans. PMID- 12766942 TI - Liberating genetic variance through sex. AB - Genetic variation in fitness is the fundamental prerequisite for adaptive evolutionary change. If there is no variation in survival and reproduction or if this variation has no genetic basis, then the composition of a population will not evolve over time. Consequently, the factors influencing genetic variation in fitness have received close attention from evolutionary biologists. One key factor is the mode of reproduction. Indeed, it has long been thought that sex enhances fitness variation and that this explains the ubiquity of sexual reproduction among eukaryotes. Nevertheless, theoretical studies have demonstrated that sex need not always increase genetic variation in fitness. In particular, if fitness interactions among beneficial alleles (epistasis) are positive, sex can reduce genetic variance in fitness. Empirical data have been sorely needed to settle the issue of whether sex does enhance fitness variation. A recent flurry of studies[1-4] has demonstrated that sex and recombination do dramatically increase genetic variation in fitness and consequently the rate of adaptive evolution. Interpreted in light of evolutionary theory, these studies rule out positive in these experiments epistasis as a major source of genetic associations. Further studies are needed, however, to tease apart other possible sources. PMID- 12766943 TI - Mitochondrial biogenesis: which part of "NO" do we understand? AB - A recent paper by Nisoli et al. [1] provides the first evidence that elevated levels of nitric oxide (NO) stimulate mitochondrial biogenesis in a number of cell lines via a soluble guanylate-cyclase-dependent signaling pathway that activates PGC1alpha (peroxisome proliferator-activated receptor gamma coactivator 1alpha), a master regulator of mitochondrial content. These results raise intriguing possibilities for a role of NO in modulating mitochondrial content in response to physiological stimuli such as exercise or cold exposure. However, whether this signaling cascade represents a widespread mechanism by which mammalian tissues regulate mitochondrial content, and how it might integrate with other pathways that control PGC1alpha expression, remain unclear. PMID- 12766944 TI - Dlg, Scribble and Lgl in cell polarity, cell proliferation and cancer. AB - Dlg (Discs large), Scrib (Scribble) and Lgl (Lethal giant larvae) are evolutionarily conserved components of a common genetic pathway that link the seemingly disparate functions of cell polarity and cell proliferation in epithelial cells. dlg, scrib and lgl have been identified as tumour suppressor genes in Drosophila, mutations of which cause similar phenotypes, involving disruption of cell polarity and neoplastic overgrowth of tissues. The molecular mechanisms by which Dlg, Scrib and Lgl proteins regulate cell proliferation are not clear, but there is some evidence that epithelial polarisation is required for this regulation. Dlg, Scrib and Lgl are highly conserved between human and Drosophila, and we discuss evidence that these proteins also play a role in cancer progression in humans. PMID- 12766945 TI - About face: signals and genes controlling jaw patterning and identity in vertebrates. AB - The embryonic vertebrate face is composed of similarly sized buds of neural crest derived mesenchyme encased in epithelium. These buds or facial prominences grow and fuse together to give the postnatal morphology characteristic of each species. Here we review the role of neural crest cells and foregut endoderm in differentiating facial features. We relate the developing facial prominences to the skeletal structure of the face and review the signals and genes that have been shown to play an important role in facial morphogenesis. We also examine two experiments one at the genetic level and one at the signal level in which transformation of facial prominences and subsequent change of jaw identity was induced. We propose that signals such as retinoids and BMPs and downstream transcription factors such as Distal-less related genes specify jaw identity. PMID- 12766946 TI - The architecture of polarized cell growth: the unique status of elongating plant cells. AB - Polarity is an inherent feature of almost all prokaryotic and eukaryotic cells. In most eukaryotic cells, growth polarity is due to the assembly of actin-based growing domains at particular locations on the cell periphery. A contrasting scenario is that growth polarity results from the establishment of non-growing domains, which are actively maintained at opposite end-poles of the cell. This latter mode of growth is common in rod-shaped bacteria and, surprisingly, also in the majority of plant cells, which elongate along the apical-basal axes of plant organs. The available data indicate that the non-growing end-pole domains of plant cells are sites of intense endocytosis and recycling. These actin-enriched end-poles serve also as signaling platforms, allowing bidirectional exchange of diverse signals along the supracellular domains of longitudinal cell files. It is proposed that these actively remodeled end-poles of elongating plant cells remotely resemble neuronal synapses. PMID- 12766947 TI - Imprinting evolution and the price of silence. AB - In contrast to the biallelic expression of most genes, expression of genes subject to genomic imprinting is monoallelic and based on the sex of the transmitting parent. Possession of only a single active allele can lead to deleterious health consequences in humans. Aberrant expression of imprinted genes, through either genetic or epigenetic alterations, can result in developmental failures, neurodevelopmental and neurobehavioral disorders and cancer. The evolutionary emergence of imprinting occurred in a common ancestor to viviparous mammals after divergence from the egg-laying monotremes. Current evidence indicates that imprinting regulation in metatherian mammals differs from that in eutherian mammals. This suggests that imprinting mechanisms are evolving from those that were established 150 million years ago. Therefore, comparing genomic sequence of imprinted domains from marsupials and eutherians with those of orthologous regions in monotremes offers a potentially powerful bioinformatics approach for identifying novel imprinted genes and their regulatory elements. Such comparative studies will also further our understanding of the molecular evolution and phylogenetic distribution of imprinted genes. PMID- 12766948 TI - Fanconi anaemia proteins: major roles in cell protection against oxidative damage. AB - Fanconi anaemia (FA) is a cancer-prone genetic disorder that is characterised by cytogenetic instability and redox abnormalities. Although rare subtypes of FA (B, D1 and D2) have been implicated in DNA repair through links with BRCA1 and BRCA2, such a role has yet to be demonstrated for gene products of the common subtypes. Instead, these products have been strongly implicated in xenobiotic metabolism and redox homeostasis through interactions of FANCC with cytochrome P-450 reductase and with glutathione S-transferase, and of FANCG with cytochrome P-450 2E1, as well as redox-dependent signalling through an interaction between FANCA and Akt kinase. We hypothesise that FA proteins act directly (via FANCC and FANCG) and indirectly (via FANCA, BRCA2 and FANCD2) with the machinery of cellular defence to modulate oxidative stress. The latter interactions may co ordinate the link between the response to DNA damage and oxidative stress parameters (3, 6-12). PMID- 12766949 TI - Species as family resemblance concepts: the (dis-)solution of the species problem? AB - The so-called "species problem" has plagued evolutionary biology since before Darwin's publication of the aptly titled Origin of Species. Many biologists think the problem is just a matter of semantics; others complain that it will not be solved until we have more empirical data. Yet, we don't seem to be able to escape discussing it and teaching seminars about it. In this paper, I briefly examine the main themes of the biological and philosophical literatures on the species problem, focusing on identifying common threads as well as relevant differences. I then argue two fundamental points. First, the species problem is not primarily an empirical one, but it is rather fraught with philosophical questions that require-but cannot be settled by-empirical evidence. Second, the (dis-)solution lies in explicitly adopting Wittgenstein's idea of "family resemblance" or cluster concepts, and to consider species as an example of such concepts. This solution has several attractive features, including bringing together apparently diverging themes of discussion among biologists and philosophers. The current proposal is conceptually independent (though not incompatible) with the pluralist approach to the species problem advocated by Mishler, Donoghue, Kitcher and Dupre, which implies that distinct aspects of the species question need to be emphasized depending on the goals of the researcher. From the biological literature, the concept of species that most closely matches the philosophical discussion presented here is Templeton's cohesion idea. PMID- 12766950 TI - The thioredoxin-like fold: hidden domains in protein disulfide isomerases and other chaperone proteins. AB - Although protein disulphide isomerase (PDI) has been known for nearly 40 years, several new PDIs have recently been described that reveal a remarkable diversity in both structure and function. This article reviews our current knowledge of the PDI family members and identifies four novel PDIs in the human genome. These include human transmembrane proteins that have C. elegans or Drosophila orthologues for which a developmental role has been proven. Their role in development, together with other functional roles for PDIs such as conferring resistance to apoptosis under hypoxia and a potential role in the oxygen-sensing apparatus are discussed. PMID- 12766951 TI - Biological activities of the shrub Salsola tuberculatiformis Botsch.: contraceptive or stress alleviator? AB - Plants belonging to the genus Salsola (Family: Chenopodiaceae) are common in the arid and semiarid regions of our planet with no less than 69 different Salsola species found in Namibia and the Republic of South Africa. This genus is used as a traditional medicine and aqueous extracts of Salsola have been used by Bushmen women as an oral contraceptive. Ingestion of the Namibian shrub Salsola tuberculatiformis Botsch. by pregnant Karakul sheep leads to prolonged gestation and fetal post-maturity and, as a result, the pelts of the new-born karakul lambs are worthless. This initiated an investigation into the active agents in the plant, using the terminal enzyme in adrenal corticosteroidogenesis, cytochrome P450-dependent 11beta-hydroxylase (P450c11), as a bioassay. Although the active fraction, S2, was extremely labile, partial structure determination suggested the presence of synephrine and a highly reactive aziridine. Therefore a more stable analogue, 2-(4-acetoxyphenyl)2-chloro-N-methylethylammonium-chloride (compound A), was synthesised, which, like the active plant extracts, inhibited adrenal steroidogenesis and acted as a contraceptive. In addition, compound A was stabilised by interaction with steroid-binding globulins in plasma thus enhancing biological activity in vivo. These findings provided explanations for the complex biological effects of the shrub as well as a new insight into the mode of action of chemically labile plant products in vivo. PMID- 12766952 TI - Creationism and the wheel. PMID- 12766953 TI - Treatment of Bowen's disease with carbon dioxide laser. PMID- 12766954 TI - Transurethral endoscopic treatment of upper urinary tract tumors using a holmium:YAG laser. AB - BACKGROUND AND OBJECTIVES: The feasibility of treating upper urinary tract tumors with a holmium:YAG (Ho:YAG) laser in transurethral endoscopy was examined. STUDY DESIGN/MATERIALS AND METHODS: Thirty-three treatments were performed on 30 patients with renal pelvic and ureteral carcinomas. After vaporization and coagulation eliminated the tumors, the surrounding mucosa was fully coagulated. Postoperative follow-up consisted of urinary cytology once a month, cystoscopy every 3 months, excretory pyelography every 6 months, and uretero-pyeloscopy every 6-12 months. RESULTS: The recurrence rate after the first treatment was 86% in the imperative indication group. The tumor-free rate (median follow-up, 37 months) in the imperative indication group was 57%. In the elective indication group, those values were 20 and 95% (median follow-up, 33 months), respectively. CONCLUSIONS: Transurethral endoscopic treatment of upper urinary tract tumors using Ho:YAG laser can be a useful method on limited cases identified into specific treatments groups combined with a strict follow-up. PMID- 12766955 TI - Preliminary study of laser welding for aortic dissection in a porcine model using a diode laser with indocyanine green. AB - BACKGROUND AND OBJECTIVES: The objective of this study was to determine whether a dissected aorta could be welded by a diode laser with a solder using an in vitro porcine aortic dissection model. STUDY DESIGN/MATERIALS AND METHODS: Porcine aortic strips were dissected into two flaps and the dissected faces were immersed in a solution of indocyanine green. The two flaps were pressed at 0.2 kg/cm2 with contact between the two immersed faces. The pressed flaps were irradiated with a diode laser (810 nm) at intensities of 170-425 W/cm2 for 8 seconds. The welded flaps were studied by light microscopy and the adhesive strengths were measured. RESULTS: The irradiated flaps were successfully welded. The breaking stress, the maximum stress recorded in a stress-strain curve, increased with increase in irradiation intensity up to 396 W/cm2 (2.7 x 10(2) mmHg) and decreased when the intensity reached 425 W/cm2. In the specimen irradiated at 396 W/cm2, the welded faces showed continuous fusion of elastin layers, while some voids were seen between the welded faces in the specimen irradiated at 425 W/cm2. CONCLUSIONS: The dissected porcine aortas were successfully welded using a laser with solder. The results suggest that the welded aorta can bear physiological blood pressure. PMID- 12766956 TI - A preliminary study of laser tissue soldering as arterial wall reinforcement in an acute experimental aneurysm model. AB - BACKGROUND AND OBJECTIVES: Aneurysm formation results from destruction of structural arterial wall connective tissue, leading to wall weakening and rupture. The purpose of this study was to demonstrate that reinforcement of the arterial wall using laser tissue soldering contributes to arterial wall stabilization and rupture prevention in an acute experimental model. STUDY DESIGN/MATERIALS AND METHODS: Elastase (10 U/mg protein, Sigma-Aldrich Co., St. Louis, MO) was applied with a fine paint brush on femoral artery segments to cause fusiform aneurysm formation. After aneurysms formed (approximately 45 minutes after treatment), elastase was rinsed out and indocyanine green (ICG) and albumin soldering mixture (2.5 mg/ml ICG in 50% albumin) was delivered to the arterial segment, followed by laser irradiation at 830 nm, (15mW output for 20 minutes). In situ pressure burst measurements were then performed. RESULTS: In situ burst pressures were > 503 mmHg for normal arteries and 181 +/- 26.0 mmHg, for Elastase treated segments. (P < 0.0001) Treatment of experimental aneurysms laser tissue soldering returned burst strengths to > 503 mmHg. CONCLUSIONS: These results indicate laser tissue soldering reinforcement of weak arterial walls, is possible and may reduce the likelihood of acute rupture. Further development of this technique for aneurysm management is warranted. PMID- 12766957 TI - Wavelength and average power density dependency of the surface modification of root dentin using an MIR-FEL. AB - BACKGROUND AND OBJECTIVES: Surface modification of root dentin by mid-infrared (MIR) pulsed-laser irradiation is one of the candidates for a novel, non-invasive treatment to prevent root surface caries. To modify root dentin effectively and non-invasively it is essential to estimate quantitatively and qualitatively the laser parameters, such as the wavelength and power density, required for surface modification. The key aspect is to bring about effective surface modification of the root dentin while minimizing the unwanted removal of the underlying dentin. STUDY DESIGN/MATERIALS AND METHODS: Using a tunable, MIR Free Electron Laser with lambda = 8.8-10.6 microm, we have investigated macroscopically the extent of the surface modification (morphological and chemical changes) of root dentin. We have obtained experimental results related to the ablation depth, the MIR absorption spectrum, and the elemental chemical composition. RESULTS: The observations showed that the surface modification of root dentin was inclined toward well recrystallized HAp-like material, leading to an increase in the acid resistance and dentinal tubule sealing. The laser parameters, at which efficient surface modification without enhanced ablation occurred, were estimated to be approximately in the wavelength region around lambda = approximately 9.0 or approximately 9.7 microm and in the average power density region of approximately 10-20 W/cm2 (resulting in total energy density and peak power density regions of approximately 1-2 kJ/cm2 and approximately 0.67-1.2 kW/cm2). CONCLUSIONS: The surface modification of root dentin strongly depends on the laser parameters applied. We conclude that the optimum wavelengths for laser treatment of root surface caries are lambda = approximately 9.0 or approximately 9.7 microm, corresponding to the absorption peak due to P-O stretching. PMID- 12766958 TI - In vivo and in vitro effects of an Er:YAG laser, a GaAlAs diode laser, and scaling and root planing on periodontally diseased root surfaces: a comparative histologic study. AB - BACKGROUND AND OBJECTIVES: The aim of the present histologic study was to compare the in vivo and in vitro effects of an erbium: yttrium, aluminum, and garnet (Er:YAG) laser (ERL), combined with a fluorescent calculus detection system, a diode laser (DL) and scaling and root planing (SRP) on periodontally diseased root surfaces. STUDY DESIGN/MATERIALS AND METHODS: Twenty-four single rooted teeth, considered for extraction due to severe periodontal destruction, were included in the study. Prior to extraction all mesial root surfaces were randomly assigned to the following treatment groups: (1) ERL combined with a calculus detection system with fluorescence induced by 655 nm InGaAsP DL radiation (160 mJ/pulse and 10 pulses/second under water irrigation) (ERL), or (2) GaAlAs DL (1.8 W, pulse/pause relation 1:10), or (3) SRP using hand instruments. Immediately after extraction, all distal root surfaces were treated with the same instruments under standardized conditions. For light microscopic investigation, a plastic embedding technique was used to cut the undecalcified roots into 30 microm thick crossections. The following parameters were recorded by on blind examiner: remaining debris, root surface morphology, and thermal side effects. RESULTS: Root surfaces instrumented with both, ERL in vivo and DL in vitro exhibited no detectable surface alterations. In contrast, ERL scaling in vitro and SRP in vivo/in vitro produced superficial microchanges in root cementum. However, irradiation with DL in vivo caused severe damages to the root surface (i.e., crater formation). There were no signs of thermal side effects in all laser treated groups. ERL provided subgingival calculus removal on a level equivalent to that provided by SRP. DL was unsuitable for calculus removal, since macroscopic inspection revealed the presence of large amounts of subgingival calculus. CONCLUSIONS: The present in vivo results showed that (i) ERL, combined with a fluorescent calculus detection system, provided a selective subgingival calculus removal on a level equivalent to that provided by SRP, and (ii) DL, using this power output, was unsuitable for calculus removal and altered the root surface in an undesirable manner. PMID- 12766959 TI - Autofluorescence characteristics of healthy oral mucosa at different anatomical sites. AB - BACKGROUND AND OBJECTIVES: Autofluorescence spectroscopy is a promising tool for oral cancer detection. Its reliability might be improved by using a reference database of spectra from healthy mucosa. We investigated the influence of anatomical location on healthy mucosa autofluorescence. STUDY DESIGN/MATERIALS AND METHODS: Spectra were recorded from 97 volunteers using seven excitation wavelengths (350-450 nm), 455-867 nm emission. We studied intensity and applied principal component analysis (PCA) with classification algorithms. Class overlap estimates were calculated. RESULTS: We observed differences in fluorescence intensity between locations. These were significant but small compared to standard deviations (SD). Normalized spectra looked similar for locations, except for the dorsal side of the tongue (DST) and the vermilion border (VB). Porphyrin like fluorescence was observed frequently, especially at DST. PCA and classification confirmed VB and DST to be spectrally distinct. The remaining locations showed large class overlaps. CONCLUSIONS: No relevant systematic spectral differences have been observed between most locations, allowing the use of one large reference database. For DST and VB separate databases are required. PMID- 12766960 TI - Measurement of the elastic modulus of rabbit nasal septal cartilage during Nd:YAG (lambda = 1.32 microm) laser irradiation. AB - BACKGROUND AND OBJECTIVES: The objective of this study was to quantitatively measure changes in the elastic moduli of rabbit nasal septal cartilage during laser heating. While the efficacy of laser cartilage reshaping has been established for use in nasal surgery, few studies have investigated the temperature-dependent viscoelastic behavior of cartilage. STUDY DESIGN/MATERIALS AND METHODS: Cyclic force versus displacement curves were generated during the Nd:YAG laser (lambda = 1.32 microm, 10 second exposure time, 21.22 W/cm2) irradiation of cartilage specimens secured in cantilevered geometry. Samples were irradiated three times with 30 second cooling intervals between each laser exposure. Measurements were recorded before, during, and after laser irradiation, and then following complete rehydration in normal saline (NS) for 1 hour at 25 degrees C. Elastic modulus was calculated assuming linear viscoelastic behavior. RESULTS: The elastic modulus in native tissue decreased during and after successive laser exposures from about 6 to 3.5 MPa. After rehydration, the modulus returned to near-baseline value. Surface temperature reached a maximum of 65 degrees C. CONCLUSIONS: The laser irradiation of cartilage using parameters similar to those used in reshaping does not produce significant irreversible changes in the mechanical properties of the tissue. Measurement of the elastic modulus is an effective means of characterizing alterations in cartilage mechanical behavior during and after laser heating. PMID- 12766961 TI - Value of endotracheal tube safety in laryngeal laser surgery. AB - BACKGROUND AND OBJECTIVES: Due to the increased popularity of laryngeal laser therapy, surgeons and anesthesiologists are inevitably confronted with questions concerning the choice of the most efficient endotracheal tube (ETT) for laryngeal laser surgery, especially with regard to possible endolaryngeal tube fires, or combustions. The purpose of this study was to determine the current practice in endolaryngeal laser surgery in Germany. STUDY DESIGN/MATERIALS AND METHODS: A questionnaire was sent to 152 ENT Departments in Germany, care was taken that the responders could send back the questionnaire anonymously. Among other questions the participants were asked for the number of lasersurgical treatments of the larynx performed in the past, the usual type of ETTs in use, whether other safety precautions were taken during CO2 laser surgery of the larynx and for intraoperative complications like tube ignition, fires or combustions. RESULTS: Eighty six of the 152 addressed ENT departments replied. In laryngeal laser surgery, 59/86 departments regularly use special laser tubes in daily routine (74.5%). In about 20,000 lasersurgical procedures, 15 incidents of ETT fire have been reported. In six of the reported 15 cases a tube fire occurred despite the fact that special laser tubes had been utilized. CONCLUSIONS: The present study could demonstrate that the use of special laser tubes does not necessarily protect against ETT fire. Thus, even when using special laser tubes other safety measures should be taken. In view of the maximum safety for the patient it has to be stated, that the safety during surgery correlates definitely with the experience of the surgeon. The weakest point of ETTs is usually situated in the cuff region. PMID- 12766962 TI - Use of Q-switched ruby laser in the treatment of nevus of ota in different age groups. AB - BACKGROUND AND OBJECTIVES: Nevus of Ota is a form of dermal melanocytic hamartoma that appears as a bluish discoloration in the trigeminal region. Although Q switched lasers provide effective treatment, the appropriate age at which to start that treatment is not known. Our aim is to compare the clinical efficacy and safety of Q-switched ruby laser (QSRL) in the treatment of nevus of Ota in different age groups. STUDY DESIGN/MATERIALS AND METHODS: Our study included 46 children and 107 adults with nevus of Ota, which had been treated with QSRL that achieved excellent (75% or more) to complete response. The laser parameters were 694-nm wavelength, 30 nanoseconds pulse duration, 4-mm spot size, and 5-7 J/cm2 fluence at 3-4 month intervals. We assessed the mean number of treatment sessions and the degree of complications in the two age groups of patients. RESULTS: The mean number of treatment sessions to achieve significance to complete clearing was 3.5 for the younger age group and 5.9 for the older age group (P = 0.0001). The complication rate for the younger age group was 4.8% as compared to 22.4% for the older age group. CONCLUSIONS: The use of QSRL for the treatment of nevus of Ota in children can achieve an excellent result in fewer sessions and at a lower complication rate than later treatment. The risk of recurrence is a concern, however, and further long-term study is necessary to address this issue. PMID- 12766963 TI - Leg ulceration after pulsed dye laser treatment of a vascular malformation. AB - BACKGROUND AND OBJECTIVES: The pulse dye laser (PDL) is a well-established treatment for port wine stains (PWSs) and is generally accepted to be safe, effective, and results in fading in the majority of patients. Significant side effects such as scarring are extremely rare. To date ulceration has not been reported. STUDY DESIGN/PATIENTS AND METHODS: We report a 22-year-old male who presented with a large vascular malformation on his lower leg with both superficial capillary and deeper venous components, which on venography and ultrasound showed abnormal architecture and reflux into the superficial collecting system. He underwent a series of test areas with the PDL on the lower leg. RESULTS: Ulceration developed in multiple sites after treatment as well as significant symptoms and signs of varicose eczema, which took at least 2 months to heal despite appropriate topical treatments and compression. CONCLUSIONS: Patients with vascular malformations of the lower leg with deeper venous components should be warned about the development of ulceration and treated with caution. PMID- 12766964 TI - Hair removal with long pulsed diode lasers: a comparison between two systems with different pulse structures. AB - BACKGROUND AND OBJECTIVES: The aim was to study hair removal efficacy, and possible side effects of two commercially available long pulsed diode lasers. The radiant exposure was selected to a value of 35 J/cm2, which is frequently used in the clinic in accordance with manufacturer's recommendations. STUDY DESIGN/MATERIALS AND METHODS: A prospective clinical study was performed on twenty-nine patients with hair color ranging from light brown to black on the upper lip. One half of the upper lip was randomly selected for treatment with the MedioStar laser; the contralateral half of the lip was treated with the LightSheer laser. Three treatments were performed at 6-8 week intervals. Percent hair reduction and acute- and long-term side effects were evaluated after treatment. RESULTS: The average hair reductions 6 months after the first treatment were 49% with the MedioStar laser and 48% with the LightSheer laser. No scarring or pigmentary change of the skin was observed after any of the treatments with either laser. However, differences in acute side effects such as degree of erythema and burned hairs were observed. CONCLUSIONS: No statistically significant differences in hair removal efficacy were observed. These results agree with mathematical modeling, which also offers a method to estimate hair removal efficacy and adverse effects for a range of hair characteristics and laser parameters. PMID- 12766965 TI - Facial rhytides--subsurfacing or resurfacing? A review. AB - STUDY DESIGN/BACKGROUND AND OBJECTIVES: Currently, ablative laser therapy (with CO2/Er:YAG lasers) is considered an effective and promising method of skin rejuvenation. The induction of collagen synthesis was observed after treatments with the CO2 laser and with the long-pulsed Er:Yag laser. In past years, the undesirable side effects and risks of these methods have led to intensified research efforts in the fields of non-ablative facial rejuvenation as well as subsurfacing by means of non-ablative laser systems and intense pulsed light systems. The objective is to achieve selective, heat-induced denaturalization of dermal collagen that leads to subsequent reactive synthesis of neocollagen but does not damage the epidermis. This article reviews the use of different types of lasers and intense pulsed light sources for the non-ablative treatment of facial rhytides. RESULTS: The results of numerous clinical and histological investigations have recently indicated that these new technologies are successful. Some studies demonstrated remarkable effects with non-ablative systems; others, however, showed only limited cosmetic improvement or none at all. CONCLUSIONS: After critical review and assessment of current literature on the treatment of rhytides, we have found that non-ablative methods do not appear to be a comparable alternative to ablative skin resurfacing in terms of their efficacy and side effects. PMID- 12766966 TI - Temperature controlled burn generation system based on a CO2 laser and a silver halide fiber optic radiometer. AB - BACKGROUND AND OBJECTIVES: Experimental animal study of burns is dependent on a reliable burn generation system. Most of the experimental systems used today are unable to produce precise partial thickness burns. This limits the ability to study minor changes associated with burn care. The aim of the study was to develop a method for generating burns with a fixed depth using a CO2 laser burn generation system. MATERIALS AND METHODS: The burn generation system was composed of two components: a burn generation device and a temperature sensing and control system. These components were designed to operate together in order to keep a constant, predetermined skin surface temperature during prolonged burn generation. One hundred thirty-eight spot burns were generated on the back of five shaved 450 g male Wistar rats. The rat skin was exposed to a 70 degrees C for 5-60 seconds. The burned areas were excised and underwent evaluation by hematoxylin-eosin-stained slide microscopy. RESULTS: A linear correlation was found between the duration of exposure and the average burn depth (r = 0.93). This correlation is represented by the equation: burn depth in millimeters = 0.012x (duration in seconds of skin exposure at 70 degrees C). CONCLUSIONS: The fiber-optic-controlled laser burn generation system studied is a reliable tool for creating partial thickness as well as full thickness skin burns in rats. PMID- 12766967 TI - Effects of linearly polarized 0.6-1.6 microM irradiation on stellate ganglion function in normal subjects and people with complex regional pain (CRPS I). AB - BACKGROUND AND OBJECTIVES: Stellate ganglion blocks are an effective but invasive treatment of upper extremity pain. Linearly polarized red and near-infrared (IR) light is promoted as a safe alternative to this procedure, but its effects are poorly established. This study was designed to assess the physiological effects of this latter approach and to quantitate its benefits in people with upper extremity pain due to Complex Regional Pain Syndrome I (CRPS I, RSD). STUDY DESIGN/MATERIALS AND METHODS: This was a two-part study. In the first phase, six adults (ages 18-60) with normal neurological examinations underwent transcutaneous irradiation of their right stellate ganglion with linearly polarized 0.6-1.6 microm light (0.92 W, 88.3 J). Phase two consisted of a double blinded evaluation of active and placebo radiation in 12 subjects (ages 18-72) of which 6 had upper extremity CRPS I and 6 served as "normal" controls. Skin temperature, heart rate (HR), sudomotor function, and vasomotor tone were monitored before, during, and for 30 minutes following irradiation. Analgesic and sensory effects were assessed over the same period as well as 1 and 2 weeks later. RESULTS: Three of six subjects with CRPS I and no control subjects experienced a sensation of warmth following active irradiation (P = 0.025). Two of the CRPS I subjects reported a >50% pain reduction. However, four noted minimal or no change and improvement did not reach statistical significance for the group as a whole. No statistically significant changes in autonomic function were noted. There were no adverse consequences. CONCLUSIONS: Irradiation is well tolerated. There is a suggestion in this small study that treatment is beneficial and that its benefits are not dependent on changes in sympathetic tone. Further evaluation is warranted. PMID- 12766968 TI - Analysis of P-glycoprotein-mediated membrane transport in human peripheral blood lymphocytes using the UIC2 shift assay. AB - BACKGROUND: During transport-associated adenosine triphosphate hydrolysis, P glycoprotein (Pgp) undergoes conformation transitions detected by UIC2, a functional anti-Pgp monoclonal antibody. A newly developed UIC2 shift assay is based on increased UIC2 reactivity in the presence of Pgp substrates. All peripheral blood leukocytes express low Pgp levels. The existing antibody-based detection methods are limited in their sensitivity and require additional techniques to simultaneously analyze Pgp expression and efflux, making it difficult to ascertain the physiologic role of Pgp-mediated transport. METHODS: We validated the UIC2 shift assay against UIC2 immunostaining and DiOC(2) efflux. The UIC2 shift assay was then used to characterize Pgp functional expression and its physiologic substrates in peripheral blood leukocytes. RESULTS: A strong correlation was observed between the UIC2 shift assay versus immunostaining and dye efflux tests. The UIC2 shift assay showed improved sensitivity (compared with conventional UIC2 staining) and allowed for simultaneous detection of Pgp expression and function. Using this assay, we identified several new Pgp substrates, including monensin and retinol, and confirmed that interleukin-2 and interferon-gamma can be transported by Pgp. CONCLUSIONS: Our findings validate the use of the UIC2 shift assay in MDR1 detection and support the idea that Pgp plays a physiologic role in immunoregulation. PMID- 12766969 TI - New flow cytometric method to quantify the inhibition of enteropathogenic Escherichia coli adhesion by anti-adhesin antibodies. AB - BACKGROUND: Pathogenesis of enteropathogenic Escherichia coli (EPEC) infections can be divided in three stages. The first one is the intestinal colonization mediated by bacterial adhesins. The second and third stages are characterized by an intimate attachment of bacteria to the enterocytes. Little information is available on the specific immune response against EPEC. Here, we describe and validate a new approach to quantify the function of anti-EPEC adhesin antibodies (Abs). METHODS: We developed a new method to quantify the function of anti adhesin Abs by flow cytometry. We used pEGFP-E22 (a rabbit EPEC E22 strain expressing the GFP protein) and HeLa cells. The adhesion of E22 bacteria to HeLa cells is mediated by AF/R2, the specific E22 adhesin. We performed short-time interaction (30 min) between pEGFP-E22 and HeLa cells. After extensive washes, 10,000 HeLa cells were acquired by flow cytometry and bacterial adhesion was quantified. Different sera were used to inhibit bacterial adhesion and recombinant MPB-Afr2G (Afr2G is the main AF/R2 subunit) was also tested in this system. RESULTS: We first verified that GFP expression by E22 did not modify bacterial adhesion. We then showed that this flow cytometry approach allowed easy quantification of bacterial adhesion and inhibition mediated by a specific anti AF/R2 serum. Moreover, recombinant AF/R2 protein reversed the effect of the anti AF/R2 serum. Finally, we validated our method using sera from E22 orally infected rabbits. We detected and quantified with this method functional specific anti AF/R2 Abs in their sera. In addition, we correlated our results with an anti AF/R2 enzyme-linked immunosorbent assay. CONCLUSIONS: We have developed a new method to detect and quantify specific anti-EPEC adhesin Abs by flow cytometry. This method is easy to use and highly reproducible. Its development could be extended to the search of specific anti-adhesin Abs in human EPEC infections. PMID- 12766970 TI - Serodiagnosis of human plague by a combination of immunomagnetic separation and flow cytometry. AB - BACKGROUND: Plague is a severe, highly communicable bacterial disease caused by Yersinia pestis. It is still endemic in more than 20 countries worldwide. Although known as a devastating disease for centuries, laboratory confirmation of clinical suspected cases is still problematic. No standardized and internationally approved test system is commercially available. The aim of this study was the introduction and evaluation of a combination of immunomagnetic separation and flow cytometry for the serodiagnosis of human plague. METHODS: Paramagnetic polystyrene beads were coated with purified F1 capsular antigen (F1 CA) and reacted with sera from plague patients, from 26 laboratory personnel vaccinated against plague and from 102 healthy blood donors (HBD). After incubation with fluorescein isothiocyanate-conjugated anti-human rabbit IgG, particle-associated fluorescence was detected by flow cytometry. RESULTS: Anti-F1 CA antibodies could be demonstrated in all patients with bacteriologically confirmed plague and in 22 sera (84.6%) from vaccinees. Only one serum in the HBD group showed a weakly positive reaction. The total assay time was less than 2 h. CONCLUSIONS: Compared with a recently published combination of an anti-F1 CA enzyme-linked immunosorbent assay (ELISA) and immunoblot, the new assay showed the same sensitivity as the ELISA and almost the same specificity (99.0 versus 100%) as the immunoblot. Allowing a rapid, reliable, and quantitative analysis, immunomagnetic separation combined with flow cytometry might replace both conventional immunoassays. PMID- 12766971 TI - Human peripheral B cells: a different cytometric point of view. AB - BACKGROUND: Human peripheral B lymphocytes, analyzed by current flow cytometers, frequently show complex patterns of morphological and fluorescence signals. However, fluorescence intensity values are commonly reported without any correlation to the cell surface area. We propose a different approach, based on the evaluation of the ratio of phenotype fluorescence intensity to forward scatter intensity, to determine the apparent fluorescence density of surface molecules. METHODS: Starting from list mode acquired data, and after logical gating of live B cells, the analytical procedure suggests a serial scanning of the FSC versus SSC plot to obtain apparent fluorescence density of progressively larger cells. RESULTS: This method, applied to normal human peripheral B lymphocytes, was able to detect the presence of steady and modulated (with respect to cell size) fluorescence densities for a variety of surface molecules. B cells from patients with B cell disorders displayed interesting alterations of the phenotype density values and distributions. CONCLUSIONS: Our preliminary data show that, in human B cell cytometry, the apparent fluorescence density based method allows one to recognize variations in fluorescence intensities solely due to cell size differences and to disclose patterns of expression not detectable by the conventional intensity based approach. PMID- 12766972 TI - Effects of resolution reduction on data analysis. AB - BACKGROUND: There is often a need in flow cytometry to display and analyze histograms at resolutions lower than those native to the data. It is common, for example, to analyze DNA histograms at 256-channel resolution, even though the data were acquired at 1,024 channels or more. The most common method for reducing resolution, referred to as the consecutive summation (CS) method, can introduce distortions into the shape of histograms. Peaks that were symmetric in the original data can become skewed in the reduced-resolution histogram. Data analysis can be negatively affected by the distortions produced by reducing the histogram resolution. An alternative technique for reducing histogram resolution, the unbiased summation (US) method, minimizes shape distortion. This paper describes the US method and examines the benefits it provides in the analysis of DNA histograms. METHODS: Reduced chi-square (RCS) was used to measure the response to three experimental variables in the least-squares analysis of simulated DNA histograms. For each variable (the percentage of coefficient of variation [%CV], number of events, and mean position of the G1 distribution), a test data set of 1,000 histograms was generated at 1,024-channel resolution. Histogram resolutions were reduced with each method and then analyzed with ModFit LT cell-cycle analysis software (Verity Software House, Topsham, ME). S-phase error and processor computation time of each method also were evaluated. A Monte Carlo experiment was performed to compare CS and US methods to theoretically correct reductions. RESULTS: CS method analysis results were negatively affected by changes in %CV, number of events, and G1 peak position. The US method produced consistently lower RCS values (more accurate results) within the tested ranges. The US method eliminated bias in S-phase error and had negligible impact on analysis processing speed. It improved RCS values 44.50% on average (P < 0.0002) with actual DNA histograms. Whereas the CS method became less accurate (chi square test) as the amount of reduction increased, the US method was unaffected, producing consistently better results. CONCLUSIONS: The US method is recommended for reducing histogram resolution in modeling applications such as DNA cell-cycle analysis. It may have implications in other areas of flow cytometric data analysis. PMID- 12766975 TI - Autonomic dysfunction in peripheral nerve disease. AB - Autonomic neuropathies are inherited or acquired neuropathies in which autonomic nerve fibers are selectively or disproportionately affected. Generally, sympathetic and parasympathetic fibers are both affected but there are exceptions. Acquired cases can be autoimmune; due to diabetes, amyloidosis, drugs, or toxins; or idiopathic. Autoimmune autonomic neuropathy is often subacute, sometimes associated with a neoplasm, and associated with high titers of antibody to ganglionic nicotinic acetylcholine receptor in about half of the severe cases. The molecular basis of inherited autonomic neuropathies is better known, including recent identification of the loci and genes of hereditary sensory and autonomic neuropathies types I, III, and IV. The inherited amyloid neuropathies are due to mutations of three proteins: transthyretin, apolipoprotein A1, and gelsolin. Non-invasive autonomic testing complements clinical and electrophysiological characterization of the autonomic neuropathies. PMID- 12766974 TI - Desminopathies: good stuff lost, garbage gained, or the trashman misdirected? PMID- 12766976 TI - Skeletal muscle disorders associated with selenium deficiency in humans. AB - Skeletal muscle disorders manifested by muscle pain, fatigue, proximal weakness, and serum creatine kinase (CK) elevation have been reported in patients with selenium deficiency. The object of this report was to review the conditions in which selenium deficiency is associated with human skeletal muscle disorders and to evaluate the importance of mitochondrial alterations in these disorders. A systematic literature review using the Medline database and Cochrane Library provided 38 relevant articles. The main conditions associated with selenium deficiency fell into three categories: (1) insufficient selenium intake in low soil-selenium areas; (2) parenteral or enteral nutrition, or malabsorption; and (3) chronic conditions associated with oxidative stress, such as chronic alcohol abuse and human immunodeficiency virus (HIV) infection. In low soil-selenium areas, reversibility of muscle symptoms was similar after selenium supplementation and placebo administration, suggesting a role for other factors in the development of disease. In parenteral or enteral nutrition, or malabsorption, muscle symptoms improved after selenium supplementation in 18 of 19 patients (median delay: 4 weeks). The reason that only a minority of selenium deficient patients present with skeletal muscle disorders is unclear and is possibly related to cofactors, such as viral infections and drugs. Prospective studies of selenium-deficient myopathies would be useful in critically ill patients, alcohol abusers, and HIV-infected patients. PMID- 12766977 TI - Respiratory insufficiency in desminopathy patients caused by introduction of proline residues in desmin c-terminal alpha-helical segment. AB - Mutations in desmin gene have been identified in patients with cardiac and skeletal myopathy characterized by intracytoplasmic accumulation of desmin reactive deposits and electron-dense granular aggregates. We characterized two new desminopathy families with unusual features of adult-onset, slowly progressive, diffuse skeletal myopathy and respiratory insufficiency. Progressive reduction of respiratory muscle strength became clinically detectable between the 3rd and the 8th years of illness and led to recurrent chest infections and death in one of the patients. Novel mutations, A357P and L370P, predicted to introduce proline residue into a highly conserved alpha-helical region of desmin, were identified. Proline is known to disrupt the alpha-helix. In addition, the A357P mutation distorts a unique stutter sequence that is considered to be critically important for proper filament assembly. Functional assessment in two cell-lines, one of which does and the other of which does not constitutively produce type III intermediate filaments, demonstrated the inability of mutant desmin carrying either the A357P or the L370P mutation to polymerize and form an intracellular filamentous network. The results of this study indicate that respiratory insufficiency is an intrinsic feature of disease associated with specific desmin mutations; in some patients, respiratory weakness may present as a dominant clinical manifestation and a major cause of disability and death. PMID- 12766978 TI - Motor unit number estimation in the evaluation of focal conduction block. AB - Temporal dispersion and phase cancellation limit the utility of amplitude reduction in compound muscle action potential (CMAP) as a measure of focal conduction block but may not affect motor unit number estimation (MUNE). Hence, MUNE offers the potential of a specific measure of conduction block. We investigated the role of MUNE in 11 patients with ulnar neuropathy and conduction block at the elbow and also in 8 normal subjects. MUNE failed to detect motor unit dropout in the patient group because reduced values for surface-recorded motor unit potentials (SMUPs) were obtained at proximal locations, suggesting that focal compression selectively damages larger motor axons, an hypothesis that has support from animal studies. We conclude that, because MUNE is affected by the physiological characteristics of functional axons surviving the underlying pathological process, the utility of MUNE is limited to diseases in which the expected pathology affects motor axons uniformly. PMID- 12766979 TI - Measurement of muscle contraction with ultrasound imaging. AB - To investigate the ability of ultrasonography to estimate muscle activity, we measured architectural parameters (pennation angles, fascicle lengths, and muscle thickness) of several human muscles (tibialis anterior, biceps brachii, brachialis, transversus abdominis, obliquus internus abdominis, and obliquus externus abdominis) during isometric contractions of from 0 to 100% maximal voluntary contraction (MVC). Concurrently, electromyographic (EMG) activity was measured with surface (tibialis anterior only) or fine-wire electrodes. Most architectural parameters changed markedly with contractions up to 30% MVC but changed little at higher levels of contraction. Thus, ultrasound imaging can be used to detect low levels of muscle activity but cannot discriminate between moderate and strong contractions. Ultrasound measures could reliably detect changes in EMG of as little as 4% MVC (biceps muscle thickness), 5% MVC (brachialis muscle thickness), or 9% MVC (tibialis anterior pennation angle). They were generally less sensitive to changes in abdominal muscle activity, but it was possible to reliably detect contractions of 12% MVC in transversus abdominis (muscle length) and 22% MVC in obliquus internus (muscle thickness). Obliquus externus abdominis thickness did not change consistently with muscle contraction, so ultrasound measures of thickness cannot be used to detect activity of this muscle. Ultrasound imaging can thus provide a noninvasive method of detecting isometric muscle contractions of certain individual muscles. PMID- 12766980 TI - Quantitative ultrasonography of skeletal muscles in children: normal values. AB - The purpose of this study was to establish normal values of muscle thickness, ratio of muscle thickness to subcutaneous fat thickness, and muscle echo intensity in children between 11 weeks and 16 years of age. Transverse scans of four muscles were made by standardized real-time ultrasound examination. The scans were digitized, and mean echo intensity was measured using gray-scale analysis. A multiple regression equation was used to study which independent parameter (age, height, weight, or sex) influenced the variables for each muscle. Muscle thickness depended on the child's weight. The other parameters did not significantly influence muscle thickness after correction for weight. The ratio of muscle thickness to subcutaneous fat thickness depended on age. Echo intensity showed no correlation with either of the variables. As a result, all normal values, including the equation to calculate them, are described. These normal data may help to determine the diagnostic value of muscle ultrasound in children with suspected neuromuscular disease. PMID- 12766981 TI - Quantitative skeletal muscle ultrasonography in children with suspected neuromuscular disease. AB - We determined prospectively the diagnostic value of quantitative ultrasonography in detecting neuromuscular disorders in children. Ultrasonographic scans of four muscles were made in 36 children with symptoms or signs suggestive of neuromuscular disease, such as muscle weakness and hypotonia. The muscle thickness, ratio of muscle thickness to subcutaneous fat thickness, and echo intensity were determined in each muscle. The echo intensity was measured using computer-assisted gray-scale analysis. Thirteen of the 36 patients had a neuromuscular disorder (6 a myopathy and 7 a neuropathy). Differentiation between neuromuscular diseases and nonneuromuscular diseases could be made on the basis of echo intensities with a sensitivity of 92%, a specificity of 90%, a positive predictive value of 86%, and a negative predictive value of 95%. We conclude that computer-assisted quantitative analysis of muscle echo intensity is a reliable method to discriminate between neuromuscular and nonneuromuscular diseases in children. PMID- 12766982 TI - Ankle kinematics to evaluate functional recovery in crushed rat sciatic nerve. AB - Peripheral nerve researchers frequently use the rat sciatic nerve crush model in order to test different therapeutic approaches. The purpose of this study was to determine the sequence of changes after an axonotmetic injury by means of a biomechanical model of the foot and ankle, and compare them with walking track analysis, over a fixed period of time. A kinematic analysis program was used to acquire ankle motion data for further analysis. Although repeated measures analysis of variance showed significant cumulative changes induced by the crush lesion for both ankle kinematic parameters and sciatic functional index, post-hoc multiple comparisons by the Student-Neuman-Keuls test revealed significant differences between week 0 and week 8 only for ankle kinematics. These results are of importance in showing the superiority of ankle kinematics in detecting small biomechanical deficits related to hyperexcitability of the plantarflexor muscles, in contrast with walking track analysis, which showed full motor functional recovery 8 weeks after the crush lesion. PMID- 12766983 TI - Maximal voluntary ventilation in myasthenia gravis. AB - Assessment of respiratory muscle weakness is important at all stages of myasthenia gravis. The maximal voluntary ventilation (MVV) is an objective dynamic method for measuring the working capacity of respiratory muscles. The clinical value of this method was studied in 24 newly diagnosed patients with myasthenia gravis, classified according to Osserman criteria (grades I, IIa, and IIb). The MVV values were normal in group I, whereas a characteristic "myasthenic pattern" of decremental respiratory volumes was demonstrated during MVV in group IIa and IIb patients, with or without dyspnea. Despite some limitations and lack of specificity, MVV may be a valuable tool in the assessment of respiratory dysfunction in patients with myasthenia gravis. Muscle Nerve, 27: 715-719, 2003 PMID- 12766984 TI - Motoneuron excitability and the F wave. AB - For three motoneuron pools that differ in excitability to Ia inputs [tibialis anterior (TA), abductor pollicis brevis (APB), and soleus], F-wave parameters were measured at rest, during voluntary contraction, and following prolonged vibration. There was an inverse relationship between F-wave chronodispersion and F-wave persistence at rest, and this appeared to be related to the ease of recording the H reflex for the motoneuron pool. During a steady voluntary contraction, overall F-wave activity increased in amplitude but decreased in duration for TA and APB. Following vibration of the test muscle at 50 HZ for 10 min there was a long-lasting depression of the H reflexes of TA and APB, but no significant change in F-wave measurements. These findings are consistent with the view that reflex discharges can prevent F waves in low-threshold motor units, and that chronodispersion is affected by the extent of reflex activity; that is, chronodispersion and related F-wave measurements do not measure motor properties exclusively. The findings also suggest that F waves provide a flawed measure of the excitability of the motoneuron pool. PMID- 12766985 TI - Breakdown of adenine nucleotide pool in fatiguing skeletal muscle in McArdle's disease: a noninvasive 31P-MRS and EMG study. AB - Energy metabolism and electrical muscle activity were studied in the calf muscles of 19 patients with proven McArdle's disease and in 25 healthy subjects. Phosphorus magnetic resonance spectroscopy and surface electromyography (S-EMG) were performed during two isometric muscle contractions of 3 min at 30% maximum voluntary contraction, one performed during normal perfusion and the other during applied ischemia. After about 1 min of ischemic muscle contraction in diseased muscle a significant acceleration in phosphocreatine breakdown was observed, along with a significant decrease in adenosine triphosphate. During both contractions the absence of glycolysis was shown by a significant alkalinization. Furthermore, in patients we observed a greater increase in the S-EMG amplitude than in control subjects. We conclude that early on during moderate exercise, a small number of muscle fibers reach metabolic depletion, indicated by a reduction in the adenine nucleotide pool. An increasing number of motor units, which are still in a high-energy state, are continuously recruited to compensate for muscle fatigue. This functional compartmentation may contribute to the pathophysiology of exercise intolerance in McArdle's disease. PMID- 12766986 TI - Ambulatory foot temperature measurement: a new technique in polyneuropathy evaluation. AB - Complaints of abnormal foot temperature are common among patients with polyneuropathy. However, there is no published method of ambulatory foot temperature measurement to identify possible thermoregulatory disturbances in these patients. We configured a digital electronic thermometer and thermocouple to measure and record distal foot and ambient temperatures simultaneously every minute for 24 to 48 h. Sixteen patients with polyneuropathy and 5 normal subjects were studied; 12 patients with polyneuropathy and 4 normal subjects had at least 24 h of successful recording. The data obtained from these patients were consistent and easily summarized by standard statistical methods. In the others, technical difficulties produced nonphysiological readings. In the patients with polyneuropathy, changes in foot temperature mirrored ambient temperature fluctuations more closely than in normal subjects. This technique shows promise in studying temperature regulation in the feet and may provide new insights into neuropathy-associated pain and the pathogenesis of polyneuropathy. PMID- 12766987 TI - Morphological changes in muscle tissue of patients with infantile Pompe's disease receiving enzyme replacement therapy. AB - Pompe's disease (glycogen storage disease type II) is an autosomal recessive myopathy caused by lysosomal alpha-glucosidase deficiency. Enzyme replacement therapy (ERT) is currently under development for this disease. We evaluated the morphological changes in muscle tissue of four children with infantile Pompe's disease who received recombinant human alpha-glucosidase from rabbit milk for 72 weeks. The patients were 2.5-8 months of age at entry. Prior to treatment, all patients showed lysosomal glycogen storage in skeletal and smooth muscle cells, vascular endothelium, Schwann cells, and perineurium. The first response to treatment was noticed in vascular endothelium and in peripheral nerves after 12 weeks of treatment at an enzyme dose of 15-20 mg/kg. Increasing the dose to 40 mg/kg led, after 72 weeks of treatment, to a reduction of glycogen storage and substantial improvement of muscle architecture in the least affected patient. Not all patients responded equally well, possibly due to differences in degree of glycogen storage and concomitant muscle pathology at the start of treatment. We conclude that intravenous administration of recombinant human alpha-glucosidase from rabbit milk can improve muscle morphology in classic infantile Pompe's disease when treatment is started before irreversible damage has occurred. PMID- 12766988 TI - Needle EMG of abductor hallucis and peroneus tertius in normal subjects. AB - Normal values for standard concentric-needle electromyography (EMG) of the abductor hallucis (AH) and peroneus tertius (PT) muscles have not been established to date, yet both are potentially useful muscles in the diagnosis of length-dependent peripheral nerve disorders. Forty-three normal asymptomatic subjects stratified by age underwent a standard set of nerve conduction studies and concentric-needle examination to exclude asymptomatic disease and develop normal values for the AH and the PT for various EMG parameters. Fibrillation potentials were observed in the AH in 21% of normal subjects (10% < 60 years old, 30% > 60 years old); 19% of subjects were unable to voluntarily activate the AH. All subjects could activate the PT regardless of age. No fibrillation potentials were observed in subjects < 60 years of age, whereas 9% of those older than 60 years had mild fibrillation potentials in the PT. The PT is a useful muscle in the evaluation of length-dependent peripheral neuropathy and other peripheral nerve disorders of the lower extremities, on the basis of ease of motor unit potential activation and analysis, and relative infrequency of fibrillation potentials in normal subjects. PMID- 12766989 TI - Myokymia, neuromyotonia, dermatomyositis, and voltage-gated K+ channel antibodies. AB - A young woman presented with facial myokymia in association with dermatomyositis. There was no evidence of peripheral neuropathy. Needle electromyography showed prominent myokymic discharges and brief neuromyotonic discharges in addition to many small-amplitude, short-duration motor unit potentials. Myokymia and dermatomyositis both responded to immunosuppressive treatment. The presence of antibodies to voltage-gated potassium channels and the association with dermatomyositis indicated an autoimmune cause for myokymia, which may have been due to reversible peripheral nerve hyperexcitability. PMID- 12766990 TI - Toxoplasmic myositis as a presenting manifestation of idiopathic CD4 lymphocytopenia. AB - Toxoplasma gondii encysts in skeletal muscle. Although only rarely found at muscle biopsy, this parasite has previously been regarded as a possible cause of polymyositis. We report a case of biopsy-proven toxoplasmic myositis in a non-HIV infected patient that led to recognition of idiopathic CD4 lymphocytopenia (ICL), a rare condition typically associated with opportunistic infections. Interestingly, the CD25(+) subset that corresponds to the CD4(+) regulatory T cells controlling autoimmune processes was lacking. Steroid and antiprotozoal therapy led to recovery. PMID- 12766991 TI - Time course of serum CK immediately before and after a single muscle cramp. PMID- 12766994 TI - Performance of two commercially available sequence-based HIV-1 genotyping systems for the detection of drug resistance against HIV type 1 group M subtypes. AB - The use of genotyping assays for the detection and evaluation of drug resistance mutations within the polymerase gene of human immunodeficiency virus type 1 (HIV 1) is becoming increasingly relevant in the clinical management of HIV-1 infection. However, genotypic resistance assays available currently have been optimised for genetic subtype B strains of the virus and many clinical centres are presented with strains from subtypes A, C, and D. In the present report, we compare the performance of two sequence-based commercially available kits, the ViroSeq Genotyping System (Applied Biosystems, Foster City, CA) and the TruGene HIV-1 Genotyping Kit (Visible Genetics, Toronto, Ontario) against a panel of 35 virus isolates from HIV-1 Group M (subtypes A-J). Full-length consensus sequences were generated by the ViroSeq genotyping system for 26 of 31 (83.8%) of the isolates tested, in contrast to the TruGene genotyping system, which generated 16 of 30 (53%) usable sequences overall. Overall, subtype B isolates were sequenced with a greater degree of success than non-subtype B isolates. Discrepancies were found between the consensus sequences reported by each system for each sample (mean difference 1.0%; range 0.0-3.2%), but these appeared to be random and did not affect interpretation of the major resistance codons. In addition, both systems were able to amplify template RNA from low copy viral load plasma samples (10(2)-10(3) RNA copies/ml) taken from a random selection of patient samples encompassing subtypes A-C. While the availability of these genotyping systems should facilitate studies of HIV-1 drug resistance in countries in which these subtypes are prevalent, the performance against subtypes other than B needs to be improved. PMID- 12766995 TI - Genetic analysis of HAV strains recovered from patients with acute hepatitis from Southern Italy. AB - Southern Italy is an endemic area for HAV infection contributing to the majority of Italian hepatitis A cases. Using molecular analysis, HAV strains have been classified in distinct genotypes and subgenotypes. To characterize HAV wild-type strains circulating in Southern Italy, sequence analysis of VP3-VP1 and VP1/2A junction regions of HAV isolates recovered from 25 patients with acute hepatitis during 2000 and 2001 was carried out. HAV isolates showed a degree of identity, after pairwise comparison with one another, ranging from 91.9-100% in the VP3-VP1 junction region and 89.9-100% in the VP1/2A junction region. All strains belonged to genotype I, with 84% (21/25) of samples clustering in subgenotype IA and 16% (4/25) in subgenotype IB. Cocirculation of subgenotypes IA and IB was observed among isolates from 2000, whereas all strains from 2001 were subgenotype IA. In addition, the subgenotype IA strains formed different clusters, one of which was related closely to some Cuban strains, showing a percent similarity of 98.8% in the 168-base pair segment encompassing the VP1/2A junction and the same amino acid substitution. The latter finding suggests that this subgenotype variant circulates also in the Mediterranean area. The results of the phylogenetic analysis confirm the genetic heterogeneity among HAV strains in Western Europe. PMID- 12766996 TI - Difference in prognosis between patients infected with hepatitis B virus with genotype B and those with genotype C in the Okinawa Islands: a prospective study. AB - The factors contributing to the prognosis of hepatitis B virus (HBV)- related chronic liver disease were assessed prospectively in 72 patients with chronic hepatitis B confirmed clinically and pathologically. A comparative study was undertaken between patients infected with genotype B and those with genotype C. During the follow-up period, 13 (81.3%) of 16 patients with genotype B who were initially hepatitis B e antigen (HBeAg) positive became HBeAg negative and 14 (51.9%) of 27 with genotype C became HBeAg negative. HBeAg had been cleared in 8 (61.5%) of 13 patients with genotype B within the first 2 years of the follow-up, but in only one (7.1%) of 14 with genotype C (P < 0.05). Four (11.4%) of 35 patients with genotype B had progressed to cirrhosis, whereas, 12 (32.4%) of 37 patients with genotype C progressed to cirrhosis, including two patients with hepatocellular carcinoma. Multivariate analysis showed that difference in HBV genotype influenced significantly either the clearance of HBeAg or the development of cirrhosis. In conclusion, HBeAg was cleared from sera more frequently and earlier in patients with genotype B compared with those with genotype C, and development of cirrhosis occurred less frequently in patients with genotype B compared with those with genotype C. Thus, HBV genotypes may influence the prognosis of HBV-related chronic liver disease. PMID- 12766997 TI - HBe antigen loss during lamivudine therapy is not caused by mutations in precore and core promoter genes in patients with chronic hepatitis B. AB - HBe antigen (HBeAg) loss or seroconversion can occur during lamivudine therapy. The purpose of this study was to analyze nucleotide sequences in precore and core promoter regions, and examine the influence of mutations in these regions on the disappearance of HBeAg during lamivudine therapy. Serial serum samples were obtained from 51 patients (HBeAg loss in 26 patients) at commencement of therapy (baseline) and after 1 year of lamivudine therapy. Serum samples were amplified with PCR and nucleotide sequences of HBV were analyzed. At baseline, a precore stop codon mutation (A1896) was identified in 8 of 26 HBeAg loss patients and in 8 of 25 HBeAg non-loss patients. At 1 year, precore mutation was observed in 4 of 14 patients analyzed who showed HBeAg loss. At 1 year, however, a precore mutation was observed also in 3 of 9 analyzed patients who showed no HBeAg loss. Core promoter mutations were noted in 21 of 26 HBeAg loss patients and in 20 of 25 HBeAg non-loss patients. At 1 year, core promoter mutations were noted in 11 of 14 HBeAg loss patients and in 8 of 9 HBeAg non-loss patients. Our data suggested that during lamivudine therapy, core promoter and precore mutations do not influence HBeAg loss or seroconversion but may reduce the viral level upon HBeAg loss or seroconversion. PMID- 12766998 TI - Characterisation of the differences between hepatitis C virus genotype 3 and 1 glycoproteins. AB - Sequence variation in the envelope E1 and E2 glycoproteins of hepatitis C virus (HCV) could account for differences in disease pathogenesis in patients infected with different genotypes. A cDNA encoding the structural region of the hepatitis C polyprotein was constructed to match the majority sequence of viral RNA extracted from a patient infected with genotype 3a (designated strain HCV3a-Gla). The principal differences predicted between E2 of HCV3a-Gla and the corresponding H77c genotype 1a protein were that the former contained six more amino acids (361 vs. 355), but it had one fewer glycosylation site. Expression studies showed that, in common with the H77c glycoproteins, E1 and E2 from HCV3a-Gla localised to the endoplasmic reticulum (ER) membrane in both Huh-7 and BHK tissue culture cells and interacted to form native complexes. Analysis of the cross-reactivity of antibodies raised against glycoproteins of genotype 1a strains showed that three of five monoclonal antibodies that recognise linear epitopes were able to detect E2 from strain HCV3a-Gla. However, neither conformational E2 antibodies nor antibodies raised against E1 were able to detect the HCV3a-Gla glycoproteins. In receptor binding assays, E2 of HCV3a-Gla consistently failed to bind CD81, a putative cell receptor for HCV. Absence of binding to CD81 and lack of recognition by most antibodies raised to genotype 1a glycoproteins indicate important differences between these glycoproteins representative of genotypes 3a and 1a. These may be pertinent to the differences in response to interferon therapy and the prevalence of steatosis reported in patients infected with these genotypes. PMID- 12766999 TI - Clinical features and progression of perinatally acquired hepatitis C virus infection. AB - The purpose of this prospective-retrospective study was to provide information about the clinical features and progression of hepatitis C virus (HCV) infection transmitted perinatally. Seventy children born to HCV infected woman were enrolled consecutively in five European centers between 1990 and 1999, provided they had HCV RNA in the serum during the first year of life and/or were still anti-HCV positive at 18 months. Sixty-two infants were followed up to 24 months of age or more (range, 24 months-11 years; average, 4.8 +/- 2.3 years). A wide range of ALT elevation was observed in 93% of the infants in the first year of life. During the follow-up, a sustained ALT normalization with loss of HCV RNA was seen in 12/62 (19%) of the children within 30 months of life; 66% of the infants had developed an ALT peak greater than 5x normal at onset (vs. 28% of children with persistent viremia; P < 0.05), and 50% had HCV genotype 3 (vs. 17% of viremic children). Conversely the cumulative probability of chronic progression was 81%. Chronic infection was asymptomatic and liver disease was mild in all 11 children who underwent a biopsy. In conclusion the early stage of acquired perinatally HCV infection is characterized by a wide range of ALT abnormalities, suggesting the interaction of multiple host and virus factors. The chronic progression rate of infection is high, but the associated liver disease is usually mild. High ALT levels at onset seem to offer greater opportunity of biochemical remission and loss of viremia during follow-up. PMID- 12767000 TI - Natural histories of hepatitis C virus infection in men and women simulated by the Markov model. AB - The Markov model was introduced to simulate natural histories of hepatitis C virus (HCV) infection in men and women. The data set was constructed on 942 HCV carriers who were examined at least once a year without receiving antiviral therapies. Based on 2,251 patient-year data, the probabilities of transition between any two of the four clinical states, i.e., asymptomatic carrier state, chronic hepatitis, liver cirrhosis and hepatocellular carcinoma in 1 year were calculated. Hepatocellular carcinoma was defined as the absorbing state from where no transitions occur. Probability matrices thus obtained on six each subsets of HCV infection (asymptomatic carrier state, chronic hepatitis and liver cirrhosis in men and women) in their forties, fifties, and sixties, were used to simulate long-term outcomes of HCV infection. Male asymptomatic carriers aged 40 years were expected to retain the asymptomatic carrier state in 2.6%, evolve into chronic hepatitis in 48.4%, liver cirrhosis in 14.6% and hepatocellular carcinoma in 34.4% after 30 years when they reached 70 years of age, in contrast to 1.9%, 45.3%, 32.8% and 20.0%, respectively, of female asymptomatic carriers. Likewise, male patients with chronic hepatitis aged 40 years were expected to remain with chronic hepatitis in 43.8%, evolve into liver cirrhosis in 15.0% and hepatocellular carcinoma in 41.1%, contrasting with 38.9%, 32.7% and 22.0%, respectively, of female patients during 30 years. The Markov model could simulate the outcomes of 153 HCV carriers identified among blood donors after 5 years. The Markov simulation would help in assessing the long-term outcome of HCV infection and making decisions in the management of HCV carriers toward prevention of hepatocellular carcinoma. PMID- 12767001 TI - Hepatitis C virus infection among noninjecting drug users in New York City. AB - The prevalence of hepatitis C virus (HCV) infection among noninjecting drug users has been reported to be higher than in the general population, but the reasons for this observation remain unclear. Noninjecting drug users aged 15-40 years and who used drugs for no longer than 10 years were enrolled in the study. The participants were interviewed about risk behaviors and had specimens drawn for serological testing. Of 276 enrolled, 4.7% were infected with HCV. Drug users who had ever sniffed or snorted heroin in combination with cocaine were significantly more likely to be infected with HCV compared with those who never sniffed or snorted heroin with cocaine. No other drug use or sexual risk behaviors were found to be associated with HCV infection. These findings suggest that sniffing or snorting heroin with cocaine may explain the increase frequently found in HCV infection among noninjectors, but further studies are necessary. PMID- 12767002 TI - Sialic acid receptor specificity on erythrocytes affects detection of antibody to avian influenza haemagglutinin. AB - Haemagglutination-inhibition tests (HI) are used to detect increases in influenza antibody in serum. However, they are relatively insensitive for the detection of human antibody responses to avian haemagglutinin, even in the presence of high titres of neutralising antibody after confirmed infection or vaccination. Human influenza viruses bind preferentially sialic acid containing N-acetylneuraminic acid alpha2,6-galactose (SAalpha2,6Gal) linkages while avian and equine viruses bind preferentially those containing N-acetylneuraminic acid alpha2,3-galactose (SAalpha2,3Gal) linkages. Increasing the proportion of SAalpha2,3Gal linkages on the erythrocytes used, by enzymatic modification or change of species, improves the ability of erythrocytes to bind to avian influenza strains and thereby improves the sensitivity of detection of antibody to avian and equine HA in a range of mammalian and human sera using HI tests. PMID- 12767003 TI - Reconstitution of lymphocyte populations and cytomegalovirus viremia or disease after allogeneic peripheral blood stem cell transplantation. AB - Early reconstitution of lymphoid populations was monitored by immunophenotyping in 57 allogeneic peripheral blood stem cell (allo-PBSC) transplant patients either with or without cytomegalovirus (CMV) viremia or disease. Cell counts for total lymphocytes and CD4(+) T cells above the percentile 60th at day 14 postransplant were associated significantly with CMV viremia-free survival within 120 days after transplant. Recovery of total lymphocyte, CD3(+), and CD8(+) T cell counts proceeded at a more rapid rate in CMV viremic patients than in nonviremic patients, irrespective of whether preemptive treatment with ganciclovir had been prescribed. Significant expansion of CD8(+) and CD8(+) CD57(+) T-cell subsets was associated with recovery from viremia and no progression to CMV disease. Immunophenotyping may provide useful information for the clinical management of CMV infection in allo-PBSC transplant recipients. PMID- 12767004 TI - Molecular epidemiological study of dengue virus type 1 in Taiwan. AB - Taiwan has experienced several major outbreaks of dengue (DEN) virus since 1981. The predominant virus type involved has been dengue virus type one (DEN-1), which first appeared in 1987. To understand the molecular epidemiology of this virus, 15 strains of DEN-1 isolated during 1987-1991 and 1994-1995, including 11 epidemic strains, two sporadic strains, and two imported strains have been studied. Fragments of 490 nucleotides (nt) from the E/NS1 junction were amplified by reverse transcription-polymerase chain reaction and the nt sequences were determined. Of the 490 nt of the E/NS1 junction, 240 nt (nt 2282-2521) were aligned and compared. Nucleotide substitutions were found at 54 positions among 15 isolates. Most nt changes were synonymous substitutions, and only three amino acid changes were found. A total of 61 strains isolated worldwide were analyzed by the Neighbor-joining method, and separated phylogenetically into three distinct genotypes, I-III. Genotype I comprised isolates from Japan and Hawaii collected in the 1940s. Genotype II included most strains isolated from Asia in 1977-1995. Genotype III consisted of isolates from three continents in 1964-1995: Asia, the Americas, and Africa. Genotype III was divided further into two subgenotypes, IIIA and IIIB. Most recent isolates from Taiwan, except for the sporadic strain isolated in 1995, were similar genetically and have been classified as Genotype II. PMID- 12767005 TI - Latency pattern of Epstein-Barr virus and methylation status in Epstein-Barr virus-associated hemophagocytic syndrome. AB - Expression of different panels of latent gene transcripts is controlled by usage of three distinct Epstein-Barr virus (EBV) nuclear antigen (EBNA) promoters (Wp, Cp, and Qp). EBV-associated hemophagocytic syndrome, which is often a fatal disease and generally occurs after primary EBV infection, is characterized by monoclonal or oligoclonal proliferation of EBV-infected T cells. The latency pattern and EBNA promoter (Wp, Cp, and Qp) usage in EBV-infected cells from three patients with EBV-associated hemophagocytic syndrome were examined by reverse transcription-polymerase chain reaction (PCR). Three samples from the patients expressed EBER, EBNA1, EBNA2, latent membrane protein (LMP)1, and LMP2A transcripts. The transcripts of EBNA1 were initiated from not only Wp/Cp but also Qp. Lytic cycle Fp-initiated EBNA1 and EBV lytic gene BZLF1 transcripts were not detected. The methylation statuses of three EBNA promoters in three patients with EBV-associated hemophagocytic syndrome and in two patients with infectious mononucleosis were also analyzed using bisulfite PCR analysis. Wp was hypermethylated, and Qp was unmethylated in both diseases. Cp was highly methylated in EBV-associated hemophagocytic syndrome, however, whereas Cp was almost unmethylated in infectious mononucleosis. These results suggest that there may be distinct EBV-infected cell populations in EBV-associated hemophagocytic syndrome, which exhibit different patterns of EBV latent gene expression. The methylation status in Cp and phenotype of EBV-infected cells may be critical differences in EBV-associated hemophagocytic syndrome and infectious mononucleosis. PMID- 12767007 TI - Enterovirus infection and activation of human umbilical vein endothelial cells. AB - Gastrointestinal tract associated lymphoid tissue is considered to be the main replication site for enteroviruses. In order to invade tissues to reach pancreatic islets, cardiac muscles, and other secondary replication sites, the virus has to survive circulation in the blood and find a way to get through endothelial cells. In the present study, the susceptibility of human endothelial cells to infections caused by human parechovirus 1 and several prototype strains of enteroviruses, representing different species (human poliovirus, human enterovirus B and C), and acting through different receptor families was examined. Primary endothelial cells isolated from human umbilical vein by collagenase perfusion and also an established human endothelial cell line, HUVEC, were used. Primary endothelial cells were highly susceptible to several serotypes of enteroviruses (coxsackievirus A13, echoviruses 6, 7, 11, 30, and poliovirus 1). However, coxsackievirus A 9 and echovirus 1 infected only a few individual cells while human parechovirus 1 and coxsackie B viruses did not show evidence of replication in primary endothelial cells. In general, primary endothelial cells were more sensitive to infection-induced cytolytic effect than HUVEC. Activation of endothelial cells by interleukin-1beta did not change the pattern of enterovirus infection. Immunofluorescence stainings of infected primary endothelial cells showed that expression of activation markers, E-selectin, and intercellular adhesion molecule-1, was clearly increased by several virus infections and the former molecule also by exposing cells to UV-light inactivated coxsackieviruses. In contrast, human leukocyte antigen-DR expression was not increased by virus infection. PMID- 12767006 TI - Molecular typing and epidemiology of enteroviruses identified from an outbreak of aseptic meningitis in Belgium during the summer of 2000. AB - Non-polio enteroviruses are the most common cause of aseptic meningitis worldwide. From May to September 2000, a major outbreak of aseptic meningitis occurred in Belgium. Cerebrospinal fluid samples (CSF) of 122 patients were found to contain enterovirus RNA using diagnostic RT-PCR that targeted a 231-bp gene fragment in the 5' noncoding region. In addition, a molecular typing method was developed based on RT-nested PCR and sequencing directly from CSF(a) 358-bp fragment in the aminoterminal part of the VP1 capsid protein. To identify the enterovirus type, nucleotide sequences of the VP1 amplicons were compared to all the enterovirus VP1 sequences available in GenBank. Echovirus 30 (31.2%), echovirus 13 (23.8%), and echovirus 6 (20.5%) were identified most frequently during the epidemic. Coxsackievirus B5 was present in 15.6% of the samples, and could be subdivided in two distinct epidemic clusters, coxsackievirus B5a (10.7%) and B5b (4.9%). Other enteroviruses encountered were echovirus 16 (5.7%), echovirus 18 (1.6%), coxsackievirus B4 (0.8%) and echovirus 7 (0.8%). The high prevalence of echovirus 13, considered previously a rare serotype, indicates it is an emerging epidemic type. To verify the typing results and to explore further the intratypical genetic variation, phylogenetic analysis was carried out. Geographical clustering of most of the strains within each type and subtype could be observed. The RT-nested PCR strategy, carried out directly on clinical samples, is a simple and rapid method for adequate molecular typing of the Group B enteroviruses causing aseptic meningitis. PMID- 12767008 TI - Human herpesvirus 7-associated meningitis and optic neuritis in a patient after allogeneic stem cell transplantation. AB - A 9-year-old boy who received allogeneic stem cell transplantation began to vomit from day 10 after transplantation. In addition to vomiting, the patient had a fever (from day 26) and severe headache (from day 34). His cerebrospinal fluid (CSF) (day 41) demonstrated pleocytosis with an absence of leukemic cells. Although the patient's symptoms were resolved with further supportive care, abrupt onset of bilateral decreased vision occurred at day 54. He was diagnosed with bilateral optic neuritis, due to the presence of disc edema and redness. Concomitant with the occurrence of aseptic meningitis, the human herpesvirus 7 (HHV-7) antibody titer increased significantly in this patient. Although neither HHV-6 nor cytomegalovirus (CMV) DNA was detected in CSF collected at day 41, HHV 7 DNA was detected in the sample. Viral DNA was not detected in CSF collected at day 93. PMID- 12767009 TI - Interaction of human herpesvirus 6 with human CD34 positive cells. AB - We reported previously that human herpesvirus 6 (HHV-6) suppresses hematopoietic colony formation of erythroid (BFU-E), granulocyte/macrophage (CFU-GM), and megakaryocyte (CFU-Meg) lineages in vitro. Here we describe the interaction between HHV-6 and human CD34+ cells, which are a major source of hematopoietic progenitor cells. CD34+ cells were immunomagnetically isolated from cord blood mononuclear cells using anti-CD34+ antibodies coated onto either Dynabeads trade mark or MACS beads. The CD34+ population selected with Dynabeads showed a broad range of fluorescence. The population selected with MACS beads showed a narrow range of fluorescence. After infection with HHV-6, two transcripts of the immediate early genes were detected with both cell populations. HHV-6 suppressed colony formation of BFU-E, CFU-GM, and CFU-Meg. HHV-6 suppressed cell growth after 3 to 7 days culture in the presence of thrombopoietin (TPO). More differentiated CD34+ cells were more susceptible to the effects of HHV-6. These data indicate that the targets for hematopoietic suppression by HHV-6 are the differentiated cells. PMID- 12767010 TI - Human herpesvirus-6 modulates RANTES production in primary human endothelial cell cultures. AB - Human herpesvirus 6 (HHV6) is a beta-herpesvirus capable of infecting several cell types from different origins. HHV6 is the etiological agent of exantem subitum and has been associated with several diseases, all characterized by an inflammatory response triggered by chemokines. We show that strain U1102 of HHV6 is able to infect persistently human endothelial cells obtained from umbilical veins, adult aorta and adult heart microvessels, without apparent cytopathic effect. Analysis by in situ PCR showed that HHV6 sequences were present in 20% of HUVEC, 10% of aortic, and 1% of heart microvascular endothelial cells. Regardless of endothelial cell origin, HHV6 infection induced de novo synthesis of the RANTES CC-chemokine. It was found, however, that microvascular endothelial cells, despite their lower susceptibility to HHV6 infection, showed the highest RANTES expression. Chemokine production occurred also in the absence of viral DNA synthesis. Furthermore, RANTES synthesis required an active viral genome, as UV inactivated HHV6 infection of endothelial cells did not lead to chemokine production. We investigated the expression of HHV6 U51 gene, which encodes a chemokine receptor that is already known to sequester and down modulate RANTES in epithelial cells. HHV6-infected endothelial cells co-expressed RANTES and U51 mRNAs starting from 12 hr up to 48 hr post-infection. Then, RANTES transcripts disappeared whereas U51 messages continued to be expressed. In conclusion, this study highlights the major role of HHV6 in endothelial cell biology and the development of inflammatory processes. PMID- 12767011 TI - Virology of infantile chronic recurrent parotitis in Santiago de Chile. AB - Infantile chronic recurrent parotitis (ICRP) has been attributed to multiple causes, including viral infections, and therefore its treatment remains empirical. Our aim was to evaluate the involvement of respiratory and oropharyngeal viruses in acute episodes of ICRP. Seventy children were studied, 50 patients and 20 age-matched controls, in a 2-year follow-up study. Saliva samples were taken from the parotid duct and analyzed by viral isolation and immunofluorescence for adenovirus (Ad), respiratory sincitial virus (RSV), parainfluenza virus (PI), influenza virus (Flu), Cytomegalovirus (CMV), and herpes simplex virus (HSV). Paired sera samples were tested by ELISA for anti Epstein-Barr virus (EBV) IgG and anti-mumps IgM and IgG. Viral infections were detected in 7/50 (14%) cases of the ICRP group: one CMV; 2 Enteroviruses isolated in human embryonic lung fibroblast cells; 1 Flu A; and 3 mumps virus. No EBV seroconversions were detected. In the control group, 2 out of the 20 children had an asymptomatic mumps positive IgM titer. Our data indicate that the main respiratory and oropharyngeal viruses are not the cause of acute episodes of ICRP in Chilean children. PMID- 12767012 TI - Molecular characterization of influenza B viruses circulating in northern Italy during the 2001-2002 epidemic season. AB - During the 2001-2002 influenza season, virological surveillance highlighted the predominant circulation of B viruses (86% of isolates) in Italy, in contrast to many other countries in Europe and North America where AH3N2 viruses were isolated most frequently, and in contrast to the infrequent isolation of B viruses in Italy during the previous two years. Associated with this predominance of influenza B was the re-emergence of B/Victoria/2/87-lineage viruses, closely related to B viruses prevalent during the 1980s, which are distinct antigenically and genetically from circulating B/Sichuan/379/99-like viruses of the B/Yamagata/16/88 lineage, which predominated in most parts of the world during the last 10 years. Ninety-four viruses isolated in two regions of northern Italy were characterized, 50 by direct sequencing of haemagglutinin (HA). Viruses of both Victoria and Yamagata lineages co-circulated throughout the 12 weeks of the influenza season. The HAs of the Yamagata-lineage viruses were heterogeneous and comprised two sublineages, represented by B/Sichuan/379/99 and B/Harbin/7/94, whereas the Victoria-lineage viruses were more homogeneous and closely related to B/Hong Kong/330/01, the current prototype vaccine strain. The antigenic and genetic characteristics of the viruses correlated with certain epidemiological features. In particular, the low age (<14 years) of individuals infected with B/Hong Kong/330/01-like viruses is likely to reflect the greater susceptibility of the youngest cohort, due to lack of previous exposure to Victoria-lineage viruses, and is consistent with the conclusion that vaccination with a B/Sichuan/379/99-like virus would give poor protection against infection with B/Hong Kong/330/01-like (Victoria-lineage) viruses. PMID- 12767013 TI - Manganese superoxide dismutase induction during measles virus infection. AB - Measles virus infection of B-cells results in marked alterations in proliferation and immunoglobulin production. Very little is known about the changes of gene expression, if any, during acute measles virus infection. To elucidate cellular genes that are induced during measles virus infection, we carried out a subtraction technique, representational differential analysis. The mitochondrial protein, manganese superoxide dismutase (MnSOD), was upregulated in B-cells during measles virus infection. Although measles virus-infected B-cells did not secrete MnSOD into the environment, it was found, using an MnSOD mimetic, that intracellular MnSOD did inhibit proliferation of the B-cells. MnSOD also decreases the titer of virus produced from infected cells. Therefore, MnSOD seems to play a role in the alteration of immune function seen upon infection of B cells with measles virus. PMID- 12767014 TI - Ex vivo cytokine responses against parvovirus B19 antigens in previously infected pregnant women. AB - Parvovirus B19 infection is a significant cause of fetal death. The aim of this study was to investigate the role of maternal immune status in modulating susceptibility to fetal B19 infection. Peripheral blood was obtained from pregnant women (n = 199) with no clinical evidence of recent B19 infection. Evaluation of ex vivo T cell responses from 149/199 individuals showed significantly higher interferon-gamma levels for seropositive individuals following VP1 (268 +/- 36 versus 103 +/- 19 pg/ml; P = 0.003) and VP2 (242 +/- 42 versus 91 +/- 16 pg/ml; P = 0.01) antigen stimulation. Significantly higher levels of interleukin-2 were also observed in seropositive individuals following both VP1 (P = 0.0003) and VP2 (P = 0.0005) stimulation. The observed Th1 cellular response is lower than that documented previously for non-pregnant individuals and strongly suggests that diminution of the maternal anti-viral immune response may increase susceptibility to fetal B19 infection. PMID- 12767015 TI - Detection of human respiratory syncytial virus sequences in peripheral blood mononuclear cells. AB - Peripheral blood mononuclear cells (PBMC) obtained from patients with lower respiratory infections were examined for the detection of human respiratory syncytial virus (RSV) sequences in the N region using the reverse transcription polymerase chain reaction (RT-PCR). RSV infection was confirmed by at least one method, i.e., virus isolation, enzyme immunoassay for viral antigen, and RT-PCR of nasopharyngeal secretions (NPS) samples. The detection rate for RSV RNA in PBMC obtained from RSV-infected patients was 40% (38/94), compared to 5% (1/20) in controls (P = 0.002). Between the groups positive (38) and negative (56) for RSV RNA in PBMC, there was no significant difference in clinical parameters. Seven patients had eight episodes of reinfection and RSV RNA was detected in 50% (4/8) during consecutive infections. Sequences of their PBMC samples were distinct from those of prototype strains of subgroup A and B. However, they were not always consistent with those of paired NPS samples. The findings suggested that RSV RNA could be detected in PBMC even during reinfection and as might have the possibility of quasispecies dynamics, reflecting the nature of RNA viruses. PMID- 12767016 TI - Studies of neutralising antibodies to SV40 in human sera. AB - It has been suggested that the low levels of antibody to the simian polyoma virus SV40 found in human sera may be linked to the use of polio vaccines. Panels of sera from areas of the world with different vaccination histories were examined to see if consistent differences could be identified. In a total of 2,054 sera from the United Kingdom, 692 from Africa and 923 from Poland taken between 1985 and 1997, the seroprevalence was generally between 3 and 5%, although exceptionally one collection from Morocco had a prevalence of 100%, and one from Poland of 0.4%. The seroprevalence showed no obvious age-dependent increase and titres were low compared to post infection animal sera. The results are consistent with previous studies and reveal no general geographically based differences related to possible differences in vaccination history, but the origin of the SV40 antibody in human sera remains to be established. PMID- 12767017 TI - Value of contrast-enhanced power Doppler sonography using a microbubble echo enhancing agent in evaluation of small breast lesions. AB - PURPOSE: The purpose of this study was to prospectively evaluate the usefulness of contrast-enhanced power Doppler sonography (PDUS) using a microbubble echo enhancing agent in differentiating between malignant and benign small breast lesions. PATIENTS AND METHODS: Between July 1, 2000, and September 30, 2001, we performed gray-scale sonographic examination of patients in whom diagnostic sonography or screening mammography had revealed solid breast lesions measuring less than 2 cm in the largest dimension. The patients were then examined on PDUS before and after injection of a microbubble contrast agent. The sonographic findings for all 3 techniques, as well as the morphologic features of the Doppler signals for each patient before and after injection of the contrast agent on PDUS, were independently assessed. Each lesion was classified as "benign" or "malignant" on the basis of specific criteria for sonographic interpretation. A hemodynamic study was performed in which time-transit profiles of the Doppler signals on contrast-enhanced PDUS were generated using a computer-assisted program, and the results for each patient were compared with the findings of a histopathologic examination of surgical specimens. RESULTS: Thirty-six patients (35 women and 1 man) with a mean age of 43.5 years (range, 18-69 years) were evaluated. The tumors ranged from 4 to 19 mm in the largest dimension. Histopathologic examination revealed that 19 tumors were benign and 17 were malignant. For morphologic diagnosis of the malignant lesions, the sensitivity of gray-scale sonography was 100%, compared with 29% for PDUS without contrast enhancement. The specificity of gray-scale sonography was 47%, compared with 74% for PDUS without contrast enhancement. Contrast-enhanced PDUS had a sensitivity of 71% and a specificity of 58%. The diagnostic accuracy was 72% for gray-scale sonography, 53% for PDUS without contrast enhancement, and 64% for contrast enhanced PDUS. The time-transit profiles of the hemodynamic study did not reveal a statistically significant difference in the accuracy rates of contrast-enhanced PDUS between benign and malignant breast lesions. CONCLUSIONS: Compared with PDUS without contrast enhancement, contrast-enhanced PDUS provides better visualization of the morphology of vascular Doppler signals that is characteristic of malignancy and therefore has a higher sensitivity and diagnostic accuracy, albeit a lower specificity. In differentiating between benign and malignant small breast lesions, contrast-enhanced PDUS can be helpful when used with gray-scale sonography and PDUS without contrast enhancement. PMID- 12767018 TI - Iliotibial band thickness: sonographic measurements in asymptomatic volunteers. AB - PURPOSE: The aims of this study were to measure the thickness of the iliotibial band (ITB) in asymptomatic adult volunteers and to determine whether there was any correlation between these measurements and the subjects' age, weight, or height. METHODS: Sonography was used to measure the ITB thickness in 31 asymptomatic volunteers (13 men and 18 women) ranging from 25 to 68 years old. Two radiologists obtained the sonographic measurements of each volunteer at the levels of the femoral condyle and the tibial condyle of both knees. Findings of fluid or bursae adjacent to the ITB were recorded. RESULTS: The mean ITB thickness was 1.95 mm +/- 0.3 mm (+/- standard deviation) at the level of the femoral condyle and 3.4 mm +/- 0.5 mm at the level of the tibial condyle. There was a statistically significant negative correlation between ITB thickness and subject age. There was no significant correlation between ITB thickness and subject weight or height. Joint fluid was present in the lateral recess of both knees in 29 volunteers (93.5%) and 1 knee joint in 2 volunteers (6.5%). Bursae were present in 3 volunteers (9.7%). CONCLUSIONS: The mean sonographic values of normal ITB thickness established using sonography may be helpful in diagnosing ITB pathology. There is no significant correlation between ITB thickness and subject weight or height, but there is a negative correlation between ITB thickness and subject age. Joint fluid in the lateral recess is present in most asymptomatic individuals, but bursae are rare. PMID- 12767019 TI - Endoscopic sonographic evaluation of the thickened gallbladder wall in patients with acute hepatitis. AB - PURPOSE: Thickening of the gallbladder wall is often observed during abdominal sonographic examination in patients with acute hepatitis. However, there is rarely an opportunity for a histopathologic analysis of these structural changes. Endoscopic sonography (EUS) can accurately delineate the structure of the gallbladder wall and therefore may be useful for visualizing changes in the gallbladder wall in patients with acute hepatitis. Hence, we prospectively studied the ability of EUS to detect specific structural changes in the gallbladder wall in patients with acute hepatitis and examined the effect of high elevation of serum liver enzyme levels on the gallbladder wall. METHODS: A study group of patients diagnosed with acute hepatitis who had gallbladder wall thickening and a control group of patients without acute hepatitis or gallbladder disease underwent EUS between May 1, 1999, and June 1, 2002. EUS was used to measure the thickness of the gallbladder wall and to visualize each of its layers. Serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels of the patients with acute hepatitis were measured at the time of the EUS examination. Statistically significant differences were determined using an independent t test and the chi-squared test. A p value of less than 0.05 was considered statistically significant. RESULTS: The acute hepatitis group comprised 28 men and 24 women with a mean age of 40.8 years. The control group comprised 25 men and 25 women with a mean age of 45.1 years. The mean gallbladder wall thickness +/- standard deviation in the acute hepatitis group (6.3 +/- 2.6 mm) was significantly greater than that in the control group (1.6 +/- 0.4 mm; p < 0.01). The mean thickness of the gallbladder wall for patients in whom both the AST and the ALT levels were 500 U/l or higher (7.0 +/- 2.6 mm) was significantly greater than that for patients with levels below 500 U/l (5.4 +/- 2.3 mm; p < 0.05). In the acute hepatitis group, EUS showed thickened, well-defined muscular and serosal layers of the gallbladder wall in 24 of the patients and a diffusely thickened gallbladder wall, in which each layer was ill defined, in the other 28 patients. The mean thickness of the gallbladder wall for patients with the pattern of ill-defined layers was significantly greater than that for the patients with the pattern of well-defined layers (p < 0.05). The pattern of ill defined layers was more common among patients in whom the serum AST and ALT levels were at least 500 U/l than among patients with levels below 500 U/l (p < 0.05). CONCLUSIONS: We propose that gallbladder wall thickening in patients with acute hepatitis is associated with prominent changes in the muscular and serosal layers. Patients with highly elevated serum liver enzyme levels are more likely to have gallbladder wall thickening and disruption of planes between the muscular and serosal layers than are patients with normal liver enzyme levels. PMID- 12767020 TI - Doppler measurement of blood flow velocities in extraocular orbital vessels in patients with obstructive sleep apnea syndrome. AB - PURPOSE: We used color Doppler sonography to determine blood flow velocities in the extraocular orbital vessels of patients with obstructive sleep apnea syndrome (OSAS) and compared the results with those of healthy control subjects without OSAS. METHODS: Patients with OSAS were classified according to the apnea-hypopnea index (AHI) as having mild OSAS (AHI < 20) or severe OSAS (AHI > or = 20). The peak systolic velocity (PSV), end-diastolic velocity (EDV), and resistance index were measured in the ophthalmic artery (OA), central retinal artery (CRA), lateral short posterior ciliary artery, and medial short posterior ciliary artery using color Doppler sonography. Only 1 eye was measured in each study participant, and right and left eyes were chosen randomly. The blood flow velocities of patients with OSAS and those of control subjects were compared with the Kruskal-Wallis test and Wilcoxon's rank-sum test. RESULTS: The study comprised 30 patients (15 with mild and 15 with severe OSAS) and 20 healthy control subjects. Blood flow velocities were higher in most measured vessels in patients with OSAS than they were in the control subjects. Among patients with mild OSAS, the PSVs and EDVs in the posterior ciliary arteries were statistically significantly higher than those of the control group (p < 0.05), but those in the OA and CRA did not differ significantly between the mild OSAS group and the control group (p > 0.05). However, as the severity of OSAS increased, the PSVs and EDVs of the OA and CRA were also affected (p < 0.05). CONCLUSIONS: Color Doppler sonographic measurements of blood flow parameters in the orbital vessels may differ significantly between patients with OSAS and those without the syndrome. Therefore, OSAS should be considered in addition to other conditions when interpreting the results of color Doppler sonography of the extraocular orbital vessels if the clinical history points toward such a diagnosis. PMID- 12767021 TI - Color Doppler imaging of orbital vessels: personal experience and literature review. AB - Many Doppler imaging studies have been performed in recent years in a large number of ocular disorders because of improvements in the Doppler equipment used for detecting and measuring the low blood-flow velocities that are a requisite for the quantitative evaluation of blood flow in the orbital vessels. The ophthalmic artery, central retinal artery and vein, posterior ciliary arteries, and the superior ophthalmic vein can be easily identified using color Doppler sonography. The changes in local blood flow in these vessels assessed by spectral analysis pulsed Doppler sonography have been used to characterize and to obtain new insights into different nontumoral vascular disorders including carotid artery stenosis, central retinal vein occlusion, giant cell arteritis, glaucoma, diabetes, fistulas, and tumoral processes of the eye and orbit. Our experience has confirmed the important role of Doppler sonography in the assessment of subclinical changes in the vascular bed, in the understanding of different processes, for following up after specific treatments, and for determining the long-term prognosis of these various conditions. PMID- 12767022 TI - Intraoperative transesophageal sonographic monitoring of tumor thrombus in the inferior vena cava during radical nephrectomy and thrombectomy for renal cell carcinoma. AB - We report the case of a 58-year-old woman with renal cell carcinoma in whom real time transesophageal sonographic monitoring of the tumor thrombus in the inferior vena cava provided dynamic information, allowing us to determine the appropriate operative procedure to use. Observation of the thrombus throughout the operation showed that mobilization of the liver resulted in compression of the inferior vena cava against the spine, increasing the risk of migration of the tumor thrombus and reinforcing the need to maintain adequate positioning of the liver to prevent such compression. The surgery was completed successfully, and the patient's postoperative course was uneventful. We recommend the use of real-time transesophageal sonographic monitoring of the tumor thrombus during such surgical procedures. PMID- 12767023 TI - Pseudoaneurysm of the gastroduodenal artery ruptured into the superior mesenteric vein in a patient with chronic pancreatitis. AB - We report the case of a patient with chronic pancreatitis that was complicated by the rare occurrence of a pseudoaneurysm of the gastroduodenal artery that ruptured into the superior mesenteric vein. The patient, a 65-year-old alcoholic man, suddenly experienced hematemesis. Gastroesophagoscopy revealed bleeding from esophageal varices; the hemorrhaging was controlled with sclerotherapy. Sonography identified a 2-cm round anechoic mass at the pancreatic head, and color Doppler imaging revealed turbulent arterial flow within the mass, leading us to the diagnosis of the pseudoaneurysm. CT and angiographic findings generally corresponded with those of sonography and confirmed our diagnosis. The pseudoaneurysm was treated successfully with embolization, and the patient was discharged 10 days after therapy. Follow-up sonography performed 2 months later confirmed the absence of blood flow within the lesion. Color Doppler sonography was very useful for diagnosing the pseudoaneurysm and planning its treatment, and we recommend its routine use in patients with chronic pancreatitis to avoid delays in diagnosing and treating such vascular complications. PMID- 12767024 TI - Pseudoaneurysm of the dorsalis pedis artery: color Doppler sonographic and angiographic findings. AB - Pseudoaneurysm of the dorsalis pedis artery is an uncommon condition that is usually caused by a traumatic injury or an iatrogenic intervention. The patient usually complains of an enlarging painless, pulsatile mass. A tentative diagnosis may be made by palpation of the pulsatile mass and detection of an associated systolic bruit. Color Doppler sonographic and arteriographic examinations can be used to confirm the diagnosis. We report the case of a 17-year-old patient with a posttraumatic pseudoaneurysm of the dorsalis pedis artery. Sonographic examinations revealed pulsatile flow into and out of a cystic structure surrounded by a thick hypoechoic wall and a "to-and-fro" pattern in the neck of the vascular mass; these findings were consistent with the diagnosis of a pseudoaneurysm. Angiography confirmed the diagnosis. The patient was treated with ligation of the artery and resection of the pseudoaneurysm. He recovered well after surgery and remained free of symptoms 3 months postoperatively. We believe that color Doppler sonography should be the procedure of choice for use in diagnosing pseudoaneurysms; arteriography can then be used to evaluate the alternative blood supply before surgery is undertaken. PMID- 12767025 TI - Superior mesenteric thrombophlebitis subsequent to acute appendicitis in an adult. PMID- 12767026 TI - Sonographic diagnosis of an unusual retained intraocular foreign body. PMID- 12767027 TI - Inguinal lymph node metastasis from nonpalpable testicular carcinoma. PMID- 12767028 TI - Blocking sensory inputs to identified antennal glomeruli selectively modifies odorant perception in Drosophila. AB - Neural coding of sensory input is a major unsolved issue in neuroscience. Current experimental methods rely on neural activity recording or visualization following sensory stimulation. Most of them, however, do not include behavioral correlates on the actual perception by the animal. We present a novel approach to address olfaction and coding in adult Drosophila. Sensory input was selectively blocked in two subsets of sensory neurons that project to different, albeit overlapping, groups of central targets, by means of tetanus toxin expressed under the control of the yeast transcription factor Gal4. Glomeruli DL1, DL2, VM1, and VM4 were tested following stimulation with benzaldehyde, ethyl acetate, propionic acid, butanol, or acetone at various concentrations. The behavioral response was found to be modified in an odorant-specific and a concentration-dependent manner. Sensory input to DL2 and, to a minor extent, VM1 and/or VM4, appear to be required for benzaldehyde perception, while acetone is processed through DL1. None of these glomeruli, however, seem necessary for butanol perception. In addition, sexual differences were observed for some stimuli. These results demonstrate the behavioral relevance of odor representation as maps of glomerular activity generated in the antennal lobes following specific sensory input. The strategy used here should be useful to characterize olfactory coding, as new and selective Gal4 lines become available. PMID- 12767029 TI - Structural and functional changes in the olfactory pathway of adult Drosophila take place at a critical age. AB - The olfactory system of several holometabolous insect species undergoes anatomical changes after eclosion of the imago, following those occurring during metamorphosis. In parallel, odor experience and learning performance also evolve with age. Here, we analyze the case of adult Drosophila females. Synaptogenesis in the antennal lobe (AL) starts in late pupa and continues during the first days of adult life, at the same time as the behavioral response to odors matures. Individual olfactory glomeruli (DM6, DM2, and V) display specific growth patterns between days 1 and 12 of adult life. Experience can modify the olfactory pathway both structurally and functionally as shown by adaptation experiments. The modifications associated with this form of nonassociative learning seem to take place at a critical age. Exposure to benzaldehyde at days 2-5 of adult life, but not at 8-11, causes behavioral adaptation as well as structural changes in DM2 and V glomeruli. Altered levels in intracellular cAMP, caused by dunce and rutabaga mutants, do not affect the normal changes in glomerular size, at least at day 6 of development, but they prevent those elicited by experience, establishing a molecular difference between glomerular changes of intrinsic versus environmental origin. Taken together, these data demonstrate an imprinting like phenomenon in the olfactory pathway of young Drosophila adults, and illustrate its glomerulus-specific dynamics. PMID- 12767030 TI - A diffusible signal attracts olfactory sensory axons toward their target in the developing brain of the moth. AB - The signals that olfactory receptor axons use to navigate to their target in the CNS are still not well understood. In the moth Manduca sexta, the primary olfactory pathway develops postembryonically, and the receptor axons navigate from an experimentally accessible sensory epithelium to the brain along a pathway long enough for detailed study of regions in which axon behavior changes. The current experiments ask whether diffusible factors contribute to receptor axon guidance. Explants were made from the antennal receptor epithelium and co cultured in a collagen gel matrix with slices of various regions of the brain. Receptor axons were attracted toward the central regions of the brain, including the protocerebrum and antennal lobe. Receptor axons growing into a slice of the most proximal region of the antennal nerve, where axon sorting normally occurs, showed no directional preference. When the antennal lobe was included in the slice, the receptor axons entering the sorting region grew directly toward the antennal lobe. Taken together with the previous in vivo experiments, the current results suggest that an attractive diffusible factor can serve as one cue to direct misrouted olfactory receptor axons toward the medial regions of the brain, where local cues guide them to the antennal lobe. They also suggest that under normal circumstances, in which the receptor axons follow a pre-existing pupal nerve to the antennal lobe, the diffusible factor emanating from the lobe acts in parallel and at short range to maintain the fidelity of the path into the antennal lobe. PMID- 12767031 TI - In vivo dynamics of CNS sensory arbor formation: a time-lapse study in the embryonic leech. AB - In the embryo of the leech Hirudo medicinalis, afferent projections of peripheral sensory neurons travel along common nerve tracts to the CNS, where they defasciculate, branch, and arborize into separate, modality-specific synaptic laminae. Previous studies have shown that this process requires, at least in part, the constitutive and then modality-specific glycosylations of tractin, a leech L1 homologue. We report here on the dynamics of growth of these projections as obtained by examining the morphology of single growing dye-filled sensory afferents as a function of time. Using 2-photon laser-scanning microscopy of the intact developing embryo, we obtained images of individual sensory projections at 3 to 30 min intervals, over several hours of growth, and at different stages of development. The time-lapse series of images revealed a highly dynamic and maturation-state-dependent pattern of growth. Upon entering the CNS, the growth cone-tipped primary axon sprouted numerous long filopodial processes, many of which appeared to undergo repeated cycles of extension and retraction. The growth cone was transformed into a sensory arbor through the formation of secondary branches that extended within the ganglionic neuropil along the anterior posterior axis of the CNS. Numerous tertiary and quaternary processes grew from these branches and also displayed cycles of extension and retraction. The motility of these higher-order branches changed with age, with younger afferents displaying higher densities and greater motility than older, more mature sensory arbors. Finally, coincident with a reduction in higher order projections was the appearance of concavolar structures on the secondary processes. Rows of these indentations suggest the formation of presynaptic en-passant specializations accompanying the developmental onset of synapse formation. PMID- 12767032 TI - Sex differences and hormone influences on tyrosine hydroxylase immunoreactive cells in the leopard frog. AB - We examined sex differences in tyrosine hydroxylase immunoreactive (TH-ir) cell populations in the preoptic area (POA), suprachiasmatic nucleus (SCN), posterior tuberculum (TP), and caudal hypothalamus (Hy) in the leopard frog (Rana pipiens), in addition to the effects of natural variation in sex steroid hormones on these same populations in both sexes. All four of these populations have been shown to be dopaminergic. Gonadal sex, androgens, and estrogen all influenced TH-ir cell numbers, but in a complicated pattern of interactions. After factoring out the effects of sex steroids by multiple regression, TH-ir cell numbers in all four areas differed between the sexes, with males having a greater number of TH-ir cells. The influence of androgens and estrogen differed by region and sex of the animals. Androgens were the main influence on TH-ir cell numbers in the POA and SCN. Plasma androgen concentrations were positively correlated with TH-ir cell numbers in both areas in males. In females, androgen concentration was negatively correlated with TH-ir cell numbers in the POA; there was no significant relationship in the SCN in females. In the more caudal populations, estrogen (E2) levels were positively correlated with TH-ir cell numbers in the TP of both males and females. In the caudal hypothalamus, E2 levels were positively correlated with TH-ir cell numbers in females, but there was no significant correlation in males. The results indicate that gonadal sex imposes a baseline sex difference in the four TH-ir (dopamine) populations, resulting in a higher number of such cells in males. Individual and sex-linked differences in gonadal steroid hormones lead to variation around this baseline condition, with androgens having a greater influence on rostral populations and estrogen on caudal populations. Last, an individual's gonadal sex determines the effect that androgens and estrogen have on each population. PMID- 12767033 TI - Estrogen alters excitability but not morphology of a sexually dimorphic neuromuscular system in adult rats. AB - In rats, motoneurons of the spinal nucleus of the bulbocavernosus (SNB) innervate the bulbocavernosus (BC) muscle, which surrounds the base of the penis. The SNB/BC is a sexually dimorphic, steroid-sensitive neuromuscular system, which is critically important in male reproductive behavior. Androgens are necessary for the development, morphology, and function of the SNB/BC system. However, estradiol (E) is also necessary for the development of the SNB/BC system, and E is capable of maintaining BC EMG activity in adulthood. In this study, we used electrophysiological and anatomical methods to examine estrogenic effects on BC EMG activity. We used a modified H-reflex testing method to investigate polysynaptic reflex characteristics in intact males, castrates, and castrates treated short term with estradiol benzoate (EB). Measures of EMG activity, response latency, and spike count were altered in castrates, but maintained in EB treated castrates to the levels of intact males. Furthermore, estrogenic effects were found in EMG activity that could be isolated to the periphery of the SNB/BC system. BC NMJ size and muscle fiber area have been demonstrated to be hormone sensitive, and we examined these for possible correlates of E's effects on BC EMG activity. BC muscles of intact males, castrates, and short-term EB-treated castrates were fixed and stained with zinc iodide and osmium tetroxide. NMJ size and muscle fiber area did not differ between groups. Together, these data suggest that E treatment results in changes in the neuromuscular periphery that maintain BC EMG activity, but this effect cannot be accounted for by changes in NMJ size or muscle fiber area. PMID- 12767034 TI - Endogenous neurotrophic factors enhance neurite growth by bag cell neurons of Aplysia. AB - Mechanisms that regulate neurite outgrowth are phylogenetically conserved, including the signaling molecules involved. Here, we describe neurotrophic effects on isolated bag cell neurons (BCNs) of substrate-bound growth factors endogenous to the sea slug Aplysia californica. Sheath cells dissociated from the pleural-visceral connectives of the Aplysia CNS and arterial cells dissociated from the anterior aorta enhance neurite outgrowth when compared to controls, i.e., BCNs grown in defined medium alone. In addition, the substrate remaining after sheath cells or arterial cells are killed significantly enhances growth, relative to all other conditions tested. For instance, primary neurites are more numerous and greater in length for BCNs cultured on substrate produced by arterial cells. These results suggest that sheath and arterial cells produce growth-promoting factors, some of which are found in the substrates produced by these cell types. Using matrix-assisted laser desorption/ionization time-of flight mass spectrometry (MALDI-TOF MS), we found that Aplysia collagen-like peptides are produced by dissociated arterial cells, and therefore likely contribute to the observed growth effects. Collagen-like peptides and other factors produced by sheath and arterial cells likely influence neurite growth in the Aplysia CNS during development, learning and memory, and regeneration after injury. PMID- 12767035 TI - Cellular physiology: challenges and opportunities for insight into biological control. PMID- 12767036 TI - Hypoxia is a major stimulator of osteoclast formation and bone resorption. AB - Hypoxia is known to act as a general stimulator of cells derived from marrow precursors. We investigated the effect of oxygen tension on the formation and function of osteoclasts, the cells responsible for bore resorption, which are of promonocytic origin. Using 7- and 13-day cultures of mouse marrow cells on ivory discs, we found that reducing oxygen tension from the ambient atmospheric level of 20% by increasing the proportion of nitrogen caused progressive increases in the formation of multinucleated osteoclasts and resorption pits. Peak effects occurred in 2% oxygen, where stimulations of resorption up to 21-fold were measured. Significant stimulations of osteoclast formation and resorption were observed even in severely hypoxic cultures gassed with 0.2% oxygen. Short-term cultures of cells disaggregated from rat bones indicated that hypoxia did not alter the resorptive activity of mature osteoclasts, but reduced their survival or adherence. In 3-day organ cultures of mouse calvarial bones, exposure to 2% oxygen resulted in maximal, fivefold stimulation of osteoclast-mediated calcium release, an effect equivalent to that of prostaglandin E(2) (PGE(2)), a reference osteolytic agent. Hypoxia also caused a moderate acidosis in calvarial cultures, presumably as a result of increased anaerobic metabolism; this observation is significant because osteoclast activation is dependent on extracellular acidification. Our experiments reveal a previously-overlooked mechanism of considerable potential importance for the regulation of bone destruction. These findings may help explain the bone loss associated with a wide range of pathological states involving local or systemic hypoxia, and emphasize the importance of the vasculature in bone. PMID- 12767037 TI - Overcoming resistance of cancer cells to apoptosis. AB - Discovery of the B cell lymphoma gene 2 (Bcl-2 gene) led to the concept that development of cancers required the simultaneous acquisition, not only of deregulated cell division, but also of resistance to programmed cell death or apoptosis. Apoptosis is arguably the common pathway to cell death resulting from a range of therapeutic initiatives, so that understanding the basis for the resistance of cancer cells to apoptosis may hold the key to development of new treatment initiatives. Much has already been learnt about the apoptotic pathways in cancer cells and proteins regulating these pathways. In most cells, apoptosis is dependent on the mitochondrial dependent pathway. This pathway is regulated by pro- and anti-apoptotic members of the Bcl-2 family, and manipulation of these proteins offers scope for a number of treatment initiatives. Effector caspases activated by the mitochondrial pathway or from death receptor signaling are under the control of the inhibitor of apoptosis protein (IAP) family. Certain proteins from mitochondrial can, however, competitively inhibit their binding to effector caspases. Information about the structure of these proteins has led to initiatives to develop therapeutic agents to block the IAP family. In addition to development of selective agents based on these two (Bcl-2 and IAP) protein families, much has been learnt about signal pathways that may regulate their activity. These in turn might provide additional approaches based on selective regulators of the signal pathways. PMID- 12767038 TI - BRCA1 gene in breast cancer. AB - The BRCA1 gene was identified and cloned in 1994 based its linkage to early onset breast cancer and breast-ovarian cancer syndromes in women. While inherited mutations of BRCA1 are responsible for about 40-45% of hereditary breast cancers, these mutations account for only 2-3% of all breast cancers, since the BRCA1 gene is rarely mutated in sporadic breast cancers. However, BRCA1 expression is frequently reduced or absent in sporadic cancers, suggesting a much wider role in mammary carcinogenesis. Since BRCA1 was cloned in 1994, its molecular function has been the subject of intense investigation. These studies have revealed multiple functions of the BRCA1 that may contribute to its tumor suppressor activity, including roles in: cell cycle progression, several highly specialized DNA repair processes, DNA damage-responsive cell cycle check-points, regulation of a set of specific transcriptional pathways, and apoptosis. Many of these functions are linked to protein:protein interactions involving different portions of the 1,863 amino acid (aa) BRCA1 protein. BRCA1 functions in cell cycle progression and the DNA damage response appear to be regulated by distinct and specific phosphorylation events, but the molecular pathways activated by these phosphorylations are only beginning to be unraveled. In addition, the reason that BRCA1 mutation carriers develop specific tumor types (breast and ovarian cancers in women and possibly prostate cancers in men) is not clearly understood. Elucidation of the precise molecular functions of the BRCA1 gene product will greatly enhance our understanding of the pathogenesis of hereditary as well as sporadic mammary carcinogenesis. PMID- 12767039 TI - Biphasic role of TGF-beta1 in signal transduction and crosstalk. AB - TGF-beta1 induces cell cycle activation in mouse embryonic fibroblasts by down regulation of p27(Kip1) but it can also induce delay of EGF-induced cell cycle activation in these cells under similar conditions. In an attempt to determine the basis for these responses, the study of early TGF-beta1-induced signal transduction pathways in the presence and absence of EGF was undertaken. It is proposed that a likely target for the inhibition by TGF-beta1 of the early EGF induced p42/p44 MAPK is at the c-Raf locus. The finding that the catalytic subunits of PKA are associated with Raf-1 within minutes following application of TGF-beta1 but not EGF in fibroblasts arrested in early G1 is suggestive of a role of PKA-Raf-1 interaction in TGF-beta1 induced delay of EGF-induced cell cycle kinetics. A model for TGF-beta1 induced translocation to the plasma membrane associated Raf-1 is proposed. Reports that Rho-like GTPase activity is critical for the activation of TGF-beta1 downstream pathways raises the question as to whether Rho proteins are involved in these observed TGF-beta1-induced responses. Post-receptor signaling mechanisms for TGF-beta1 and cross-talk with PKA-mediated pathways are examined in an effort to explain the modulation by TGF-beta1 of mitogen-induced cell proliferation in mesenchymal cells. PMID- 12767040 TI - Expression of connective tissue growth factor in bone: its role in osteoblast proliferation and differentiation in vitro and bone formation in vivo. AB - Connective tissue growth factor (CTGF) is a secreted, extracellular matrix associated signaling protein that regulates diverse cellular functions. In vivo, CTGF is expressed in many tissues with highest levels in the kidney and brain. The purpose of this study was twofold; first, to localize CTGF in normal bone in vivo during growth and repair, and second, to examine CTGF expression and function in primary osteoblast cultures in vitro and test its effect on bone formation in vivo. Northern and Western blot analyses confirmed that CTGF is expressed in normal long bones during the period of growth or modeling. In situ hybridization and immunohistochemical analysis demonstrated intense staining for CTGF mRNA and protein in osteoblasts lining metaphyseal trabeculae. Examination of CTGF expression in the fracture callus demonstrated that it was primarily localized in osteoblasts lining active, osteogenic surfaces. In primary osteoblast cultures, CTGF mRNA levels demonstrated a bimodal pattern of expression, being high during the peak of the proliferative period, abating as the cells became confluent, and increasing to peak levels and remaining high during mineralization. This pattern suggests that CTGF may play a role in osteoblast proliferation and differentiation as previously demonstrated for fibroblasts and chondrocytes. Treatment of primary osteoblast cultures with anti CTGF neutralizing antibody caused a dose-dependent inhibition of nodule formation and mineralization. Treatment of primary osteoblast cultures with recombinant CTGF (rCTGF) caused an increase in cell proliferation, alkaline phosphatase activity, and calcium deposition, thereby establishing a functional connection between CTGF and osteoblast differentiation. In vivo delivery of rCTGF into the femoral marrow cavity induced osteogenesis that was associated with increased angiogenesis. This study clearly shows that CTGF is important for osteoblast development and function both in vitro and in vivo. PMID- 12767041 TI - Codominance of cisplatin resistance in somatic cell hybrids. AB - Intrinsic or acquired resistance to cisplatin in cancer cells remains a major obstacle to successful chemotherapy. The clinically relevant genetic and molecular mechanisms of resistance have not yet been identified. Cisplatin resistant (CP-r) human KB epidermoid carcinoma cell lines (HeLa) resistant to varying levels of cisplatin after single and multiple selection steps are cross resistant to other platinum compounds and to methotrexate. Intraspecies hybrids of the sensitive and KB CP-r cells were fused with HeLa D98(OR) CP-s, hypoxanthine-aminopterin-thymidine (HAT) sensitive, ouabain resistant, to determine whether cisplatin resistance is dominant or recessive. Cell-cell hybridization between the sensitive cells and single-step or two-step KB CP-r cells both indicated codominance of cisplatin resistance compared to hybrids between sensitive cell lines (D98(OR)xKB). The hybrids between sensitive cell lines (D98xKB) and a single-step CP-r KB cell line (D98xKB-CP.5) also were cross resistant to carboplatin and methotrexate. In addition, the relatively slower growth rate of CP-r cells appears to be dominant. In the two-step CP-r KB cell line, KB-CP1, resistance is no more dominant than in the single-step CP-r KB cell line, KB-CP.5, suggesting that one of the two steps of resistance in KB-CP1 may not be dominant. These dominance data suggest that it might be possible to identify one or more genes responsible for cisplatin resistance by gene transfer from a resistant cell line to a sensitive cell line. PMID- 12767042 TI - Characterization of proliferating human skeletal muscle-derived cells in vitro: differential modulation of myoblast markers by TGF-beta2. AB - Adult human skeletal muscle-derived cells (HuSkMC) propagated in vitro are under investigation as a cell-based therapy for the treatment of myocardial infarction. We have characterized HuSkMC with respect to cell identity and state of differentiation as a prerequisite to their clinical use. Flow cytometric analysis of propagated HuSkMC revealed a population of cells that expressed the myoblast markers CD56 and desmin. The presence of myoblasts in these cultures was further confirmed by their capacity to form myotubes and increase creatine kinase activity when cultured in low serum conditions. The non-myoblast fraction of these propagated cells expressed TE7, a marker associated with the fibroblast phenotype. Spontaneous differentiation of myoblasts occurred during serial propagation of HuSkMC, as judged by myotube formation, thereby reducing the myoblast representative fraction with continued cell expansion. We examined transforming growth factor beta2 (TGF-beta2) for its utility in controlling this spontaneous differentiation of adult human myoblasts in vitro. Propagation of HuSkMC in the presence of 1 ng/ml TGF-beta2 for 5 days decreased desmin expression within the myoblast population and caused a parallel reduction of creatine kinase activity. CD56 expression was unaffected, indicating a differential regulation of these myoblast markers. The reduction in desmin expression and creatine kinase activity was, however, reversible upon the removal of TGF-beta. These data collectively indicate that TGF-beta2 restrained differentiation of adult human skeletal myoblasts during propagation without causing irreversible loss of the myoblast phenotype, demonstrating the potential utility of using TGF-beta2 during cultivation and expansion of HuSkMC intended for therapeutic implantation. PMID- 12767043 TI - Threonine 308 phosphorylated form of Akt translocates to the nucleus of PC12 cells under nerve growth factor stimulation and associates with the nuclear matrix protein nucleolin. AB - We have examined the issue of whether or not in PC12 cells it may be observed a nerve growth factor (NGF) nuclear translocation of an active (phosphorylated) Akt. Western blot analysis with antibodies to either total or phosphorylated Akt showed a maximal nuclear translocation after 15 min of NGF stimulation. NGF increased rapidly and transiently the enzymatic activity of immunoprecipitable nuclear Akt and after 45 min the values returned to a level close to the basal one. Enzyme translocation was blocked by the selective phosphoinositide 3-kinase inhibitor, LY294002. Confocal microscopy of samples stained with antibody to Akt showed an evident increase in immunostaining intensity in the nuclear interior after NGF treatment. Treatment of cells with inhibitors of protein phosphatase PP2A, calyculin A, or okadaic acid, maintained the phosphorylation levels of nuclear Akt. Immunoprecipitation experiments revealed an association between Akt and PP2A that was maximal when nuclear Akt activity was decreased. Both total and active Akt associated with the nuclear matrix and, in particular, with the protein nucleolin, with which Akt co-immunoprecipitated. These findings strongly suggest that the intranuclear translocation of active Akt is an important step in the signaling pathways elicited by the neurotrophin NGF and that the intranuclear control of Akt is achieved through the action of PP2A. PMID- 12767045 TI - Adhesion-dependent control of Akt/protein kinase B occurs at multiple levels. AB - The protein kinase Akt, also known as Protein Kinase B, has been implicated in the survival of several cell types challenged with various apoptotic stimuli. In CCL39 lung fibroblasts, apoptosis is induced by anchorage and mitogen removal. Mitogen-induced activation of Akt is highly anchorage dependent in these cells and removal of adhesion is accompanied by a rapid loss in responsiveness to soluble agonists followed by a significant decrease in Akt abundance. Loss of the protein appears to be independent of kinase activation since the expression of a constitutively active form, gag-Akt, is also dependent upon cell adhesion. Although the disappearance of Akt is coincident with the induction of programmed cell death, it cannot be fully prevented by treatment of cells with the caspase inhibitor ZVAD or by sustained activation of the anti-apoptotic Raf/ERK pathway, in cells expressing an inducible DeltaRaf-1:ER construct. In addition, a previously unrecognized decrease in Akt mRNA levels following anchorage removal occurs suggesting that anchorage-dependent transcriptional and/or post transcriptional mechanisms contribute to the adhesion-dependent regulation of Akt expression. PMID- 12767044 TI - Transgenic mice with dominant negative PKC-theta in skeletal muscle: a new model of insulin resistance and obesity. AB - Protein kinase C theta (PKC-theta) is the PKC isoform predominantly expressed in skeletal muscle, and it is supposed to mediate many signals necessary for muscle histogenesis and homeostasis, such as TGFbeta, nerve-dependent signals and insulin. To study the role of PKC-theta in these mechanisms we generated transgenic mice expressing a "kinase dead" mutant form of PKC-theta (PKC thetaK/R), working as "dominant negative," specifically in skeletal muscle. These mice are viable and fertile, however, by the 6-7 months of age, they gain weight, mainly due to visceral fat deposition. Before the onset of obesity (4 months of age), they already show increased fasting and fed insulin levels and reduced insulin-sensitivity, as measured by ipITT, but normal glucose tolerance, as measured by ipGTT. After the 6-7 months of age, transgenic mice develop hyperinsulinemia in the fasting and fed state. The ipGTT revealed in the transgenic mice both hyperglycemia and hyperinsulinemia. At the molecular level, impaired activation of the IR/IRS/PI3K pathway and a significant decrease both in the levels and in insulin-stimulated activation of the serine/threonine kinase Akt were observed. Taken together these data demonstrate that over-expression of dominant negative PKC-theta in skeletal muscle causes obesity associated to insulin resistance, as demonstrated by defective receptor and post-receptorial activation of signaling cascade. PMID- 12767046 TI - Thrombin downregulates muscle acetylcholine receptors via an IP3 signaling pathway by activating its G-protein-coupled protease-activated receptor-1. AB - Regulation of thrombin activity may be required during skeletal muscle differentiation since the thrombin tissue inhibitor protease nexin-1 appears at the myotube stage before being localized at the neuromuscular synapse. Here, we have used a model of rat fetal myotube primary cultures to study the effect of thrombin on acetylcholine receptor (AChR) expression, which is enhanced at the myotube stage. Our results show that thrombin decreases both the number of surface AChRs (AChRn) and AChR alpha-subunit gene expression. Using the agonist peptide SFLLRN, we establish that the AChRn decrease is mediated by the G protein coupled thrombin receptor "protease-activated receptor-1" (PAR-1). Moreover, the specific thrombin inhibitor hirudin increases AChRn by inhibiting the thrombin intrinsically present in the cultures. We further demonstrate that the activation of PAR-1 by thrombin induces intracellular calcium movements that are blocked by 2-APB, an inhibitor of inositol 1,4,5-triphosphate (IP3)-induced calcium release. These calcium signals are more intense in nuclei than in the cytoplasm and are consistent with the intracellular distribution of IP3 receptor that we find in the cytoplasm in a cross-striated pattern and at a high level in the nuclear envelope zone. Finally, we show that the blockade of these IP3-induced calcium signals by 2-APB prevents the AChRn decrease induced by thrombin. Our results thus demonstrate that thrombin downregulates AChR expression by activating PAR-1 and that this effect is mediated via an IP3 signaling pathway. PMID- 12767047 TI - EGF receptor-mediated, c-Src-dependent, activation of Stat5b is downregulated in mitogenically responsive hepatocytes. AB - Signal transducer and activator of transcription (STAT) proteins may be activated by epidermal growth factor (EGF), but their role in EGF receptor-mediated mitogenic signaling is not clear. We previously showed that Stat5b was activated by EGF in rat hepatocytes in primary monolayer culture. In the present study, we found that EGF induced tyrosine phosphorylation of Stat5b both on Tyr-699, which correlated with specific DNA binding activity, and also on other tyrosine residues. The Src tyrosine kinase inhibitor CGP77675 blocked the EGF-induced activation of Stat5b, but did not affect the Stat5b activation by growth hormone (GH) or prolactin (PRL). The Stat5b response to EGF was most pronounced soon (3 h) after plating (early G(1)) and at high cell density (50,000 hepatocytes per cm(2)). However, at this cell density EGF did not stimulate DNA synthesis. In hepatocytes at 24 h of culturing (mid/late G(1)) with 20,000 cells per cm(2), EGF induced strong phosphorylation of the EGF receptor, as well as Shc and ERK, and stimulated DNA synthesis, but did not activate Stat5b, although the Stat5b response to GH or PRL was retained. A strong GH-induced Stat5b activation neither influenced the DNA synthesis alone nor enhanced the mitogenic effect of EGF. The results show that EGF induces tyrosine phosphorylation and DNA-binding activity of Stat5b in a manner different from GH and PRL, apparently by a Src-dependent mechanism. The data also provide further evidence that Stat5b is not required for mitogenic signaling from the EGF receptor. PMID- 12767048 TI - Post-transcriptional expression of DMT1 in the heart of rat. AB - Non-transferrin-bound iron (NTBI) overtaken by heart cells might be a key cause leading to iron-mediated injury in heart disorders. NTBI uptake by heart cells might be mediated by divalent metal transporter 1 (DMT1). The understanding of the role of DMT1 in heart iron metabolism is fundamental for elucidating the cause resulting in excessive iron in the heart. The study was to evaluate effects of age and dietary iron on DMT1 mRNA expression and protein synthesis in rat heart. DMT1 mRNA expression was determined by RT-PCR and sequence analysis, and DMT1 protein by Western blot analysis. DMT1 mRNAs with or without iron-responsive element (IRE) both were found in rat heart. Expression of two forms of DMT1 mRNAs was the lowest at the age of post-natal day (PND) 7, and then increased with the age, reaching the highest at PND196 (non-IRE form) and PND63 (IRE form), respectively. During different ages, the levels of DMT1 (IRE) mRNA were higher than those of DMT1 (non-IRE) mRNA and were significantly correlated with the non heme iron contents in the heart. After fed a high iron for 6 weeks, the rats had a sixfold elevation in heart iron and 22% (non-IRE from) and 40% (IRE from) reduction in DMT1 protein compared to the controls. A low iron diet for 6-weeks caused cardiac hypertrophy and heart iron deficiency and also an increase in levels of two forms of DMT1 proteins. However, iron status had no significant effect on DMT1 (IRE) and DMT1 (non-IRE) mRNAs expression in the heart, although it can significantly influence heart transferrin receptor (TfR) mRNA expression. The results demonstrated that DMT1 mRNAs expression in the heart is age-dependent and that two forms of DMT1 mRNAs both are regulated by iron on the post transcriptional level only. PMID- 12767049 TI - Primary culture model of peroxisome proliferator-activated receptor gamma activity in prostate cancer cells. AB - BRL 49653 (rosiglitazone) is a thiazolidinedione anti-diabetic drug that activates the nuclear receptor, peroxisome proliferator-activated receptor gamma (PPARgamma). Pilot clinical trials have shown evidence of therapeutic activity of PPARgamma agonists against prostate cancer. To more effectively use PPARgamma ligands to treat this common and generally chemo-resistant type of cancer, it will be necessary to better understand the nature of PPARgamma activity in prostate cancer cells. Tumor suppressor effects of activation of PPARgamma may include suppression of growth and/or induction of differentiation or apoptosis. We investigated responses of primary cultures of human prostatic cancer cells to BRL 49653. PPARgamma was expressed in all of the cell strains examined. BRL 49653 caused dose- and time-dependent growth inhibition that was associated with increased expression of the transcription repressor, transforming growth factor beta-stimulated clone 22 (TSC-22), and markedly increased expression of the secretory differentiation-associated gene adipophilin. Adipocyte-type fatty acid binding protein (aFABP), neutrophil gelatinase-associated lipocalin (NGAL), glycerol kinase (GyK), and beta-catenin, which are regulated by PPARgamma ligands in certain other types of cells, were not regulated by BRL 49653 in prostate cells. Upregulation of adipophilin coincided with morphological changes and the appearance of cytoplasmic vacuoles with ultrastructural features of secondary lysosomes. These results extend previous studies with established cancer cell lines and show that PPARgamma agonists can inhibit proliferation and modulate expression of secretory-associated genes in primary cultures of prostate cancer cells, further warranting consideration of these agents as pro-differentiating chemotherapeutic or chemoprevention agents for the treatment of prostate cancer. PMID- 12767050 TI - Functional activity of serotoninergic and melatoninergic systems expressed in the skin. AB - We tested the expression of genes coding receptors of a cutaneous serotoninergic/melatoninergic system in whole human skin and in normal and pathologic cultured skin cells. Evaluation of serotonin (5HT), melatonin (MT), and melatonin-related receptors (MRR) showed expression of the isoforms 5HT2B, 5HT7, and MT1 genes in almost all the tested samples. Expression of other isoforms was less prevalent; 5HT2C, MRR, and MT2 were rarely detected. We also found novel isoforms for MT2, MRR, and 5HT2B and documented the process of RNA editing for 5HT2C. Testing for functional activity of these receptors with serotonin and melatonin (10(-14) to 10(-10) M) showed variable effects depending on cell type and culture conditions. Thus, serotonin stimulated proliferation of melanocytes in medium deprived of growth factors, while inhibiting cell growth in the presence of growth factors. Melatonin inhibited both apoptosis of HaCaT keratinocytes incubated in serum-free media, and proliferation of cells cultured in medium supplemented with serum. Melatonin also increased the numbers of viable fibroblasts incubated in serum free medium. N-acetylserotonin (NAS) and 5 methoxytryptamine (5MTT) were generally without effect on cell proliferation, with the exception of an inhibition of melanocyte proliferation at the higher 5MTT concentration of 10(-10) M. Thus, skin cells represent a true target for the products of the serotoninergic/melatoninergic cutaneous pathway with their actions modulating cell proliferation or viability. PMID- 12767051 TI - Induction of leukotriene C4 synthase after the differentiation of rat basophilic leukemia cells with retinoic acid and a low dose of actinomycin D and its suppression with methylprednisolone. AB - Leukotriene C(4) synthase (LTC(4) S) is a pivotal enzyme for generation of cysteinyl-leukotrienes (cysLTs). LTC(4) S activity in rat basophilic leukemia-1 (RBL-1) cells increased after culture in the presence of retinoic acid (RA) analogues, which was inhibited by cycloheximide or actinomycin D (ACD). Unexpectedly, the co-addition of a low dose of ACD with RA further potentiated the upregulation of the LTC(4) S activity. Daunorubicin and mitomycin C also had a similar effect. When stimulated with calcium ionophore A23187, control cells did not produce cysLTs, but RA-treated cells generated cysLTs and the co-addition of ACD further increased. While LTC(4) S mRNA and protein increased in the cells treated with RA, the co-addition of ACD further potentiated both in proportion to the LTC(4) S activity. The effect of ACD was considered to enhance the transcription rate of LTC(4) S gene, but not the mRNA-stability. The addition of methylprednisolone (MP) inhibited generation of cysLTs from the cells with A23187 stimulation and also did LTC(4) S activity, but did not inhibit 5-lipoxygenase (5 LOX). The suppression of LTC(4) S with MP showed a dependent manner on the time point and duration of MP-treatment after RA-addition which was correlated with reduction in LTC(4) S mRNA and protein. The cells cultured with RA plus ACD contained more histamine, chymase activity, and granules in the cytoplasm than the control cells, suggesting differentiation to mature mast cells. In consideration of RA-differentiation therapy, it may be of pathophysiological relevance that the antineoplastic agents potentiate RA-induced, steroid sensitive, induction of LTC(4) S in RBL-1 cells. PMID- 12767052 TI - Modulation of human fibroblast gap junction intercellular communication by hyaluronan. AB - The composition of the extracellular matrix changes during dermal repair. Initially, hyaluronan (HA) concentration is high, however, by day 3, HA is eliminated. HA optimizes collagen organization within granulation tissue. One possible mechanism of HA modulation of collagen packing is through the promotion of gap junction intercellular communication (GJIC). Gap junctions are gated channels that allow rapid intercellular communication and synchronization of coupled cell activities. The gap junction channel is composed of connexin (Cx) proteins that form a gated channel between coupled cells. HA is reported to enhance Cx43 expression in transformed fibroblasts. GJIC was quantified by the scrape loading technique and reported as a coupling index. The coupling index for human dermal fibroblasts was 4.6 +/- 0.2, while the coupling index for fibroblasts treated with HA more than doubled to 10.6 +/- 0.7. By Western blot analysis no differences were appreciated in the protein levels of Cx43 or beta catenin, a protein involved in the translocation of Cx to the cell surface. By immuno-histology Cx43 and beta-catenin were evenly distributed throughout the cell in controls, but in cells treated with HA these proteins were co-localized to the cell surface. Coupled fibroblasts are reported to enhance the organization of collagen fibrils. It is proposed that HA increases the accumulation of Cx43 and beta-catenin on the cell surface, leading to greater GJIC and enhanced collagen organization. PMID- 12767053 TI - Modulation of metabolic control by angiotensin converting enzyme (ACE) inhibition. AB - Angiotensin converting enzyme (ACE) inhibitors are a widely used intervention for blood pressure control, and are particularly beneficial in hypertensive type 2 diabetic subjects with insulin resistance. The hemodynamic effects of ACE inhibitors are associated with enhanced levels of the vasodilator bradykinin and decreased production of the vasoconstrictor and growth factor angiotensin II (ATII). In insulin-resistant conditions, ACE inhibitors can also enhance whole body glucose disposal and glucose transport activity in skeletal muscle. This review will focus on the metabolic consequences of ACE inhibition in insulin resistance. At the cellular level, ACE inhibitors acutely enhance glucose uptake in insulin-resistant skeletal muscle via two mechanisms. One mechanism involves the action of bradykinin, acting through bradykinin B(2) receptors, to increase nitric oxide (NO) production and ultimately enhance glucose transport. A second mechanism involves diminution of the inhibitory effects of ATII, acting through AT(1) receptors, on the skeletal muscle glucose transport system. The acute actions of ACE inhibitors on skeletal muscle glucose transport are associated with upregulation of insulin signaling, including enhanced IRS-1 tyrosine phosphorylation and phosphatidylinositol-3-kinase activity, and ultimately with increased cell-surface GLUT-4 glucose transporter protein. Chronic administration of ACE inhibitors or AT(1) antagonists to insulin-resistant rodents can increase protein expression of GLUT-4 in skeletal muscle and myocardium. These data support the concept that ACE inhibitors can beneficially modulate glucose control in insulin-resistant states, possibly through a NO-dependent effect of bradykinin and/or antagonism of ATII action on skeletal muscle. PMID- 12767054 TI - Effect of high phosphate concentration on osteoclast differentiation as well as bone-resorbing activity. AB - Although high inorganic phosphate (Pi) concentration in culture media directly inhibits generation of new osteoclasts and also inhibits bone resorption by mature osteoclasts, its precise mechanism and the physiological role have not been elucidated. The present study was performed to investigate these issues. Increase in extracellular Pi concentration ([Pi](e)) (2.5-4 mM) concentration dependently inhibited 1,25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3)] or parathyroid hormone (PTH)-(1-34)-induced osteoclast-like cell formation from unfractionated bone cells in the presence of stromal cells. Increase in [Pi](e) (2.5-4 mM) concentration dependently inhibited 1,25(OH)(2)D(3)-, PTH-(1-34)-, or receptor activator of NF-kappaB ligand (RANKL) and macrophage colony-stimulating factor (M CSF)-induced osteoclast-like cell formation from hemopoietic blast cells in the absence of stromal cells. Increase in [Pi](e) (2.5-4 mM) dose dependently stimulated the expression of osteoprotegerin (OPG) mRNA and increased the expression of OPG mRNA suppressed by PTH-(1-34) or 1,25(OH)(2)D(3) in unfractionated bone cells, while it did not affect RANKL mRNA. Increase in [Pi](e) (2.5-4 mM) concentration dependently inhibited the bone-resorbing activity of isolated rabbit osteoclasts. Increase in [Pi](e) (4 mM) induced the apoptosis of isolated rabbit osteoclasts while it did not affect the apoptosis of osteoclast precursor cells and mouse macrophage-like cell line C7 cells that can differentiate into osteoclasts in the presence of RANKL and M-CSF. These results indicate that increase in [Pi](e) inhibits osteoclast differentiation both by up regulating OPG expression and by direct action on osteoclast precursor cells. It is also indicated that increase in [Pi](e) inhibits osteoclastic activity at least in part by the direct induction of apoptosis of osteoclasts. PMID- 12767055 TI - 1-40 Beta-amyloid protein fragment modulates the expression of CD44 and CD71 on the astrocytoma cell line in the presence of IL1beta and TNFalpha. AB - The modulation of CD44, VCAM-1 and CD71 expression was analysed by flow cytometry in the 1321N1 astrocytoma cell line in the presence of interleukin-1beta (IL1beta), tumour necrosis factor-alpha (TNFalpha) and 1-40 or 25-35 beta-amyloid (Abeta) fragments. The percentage of 1321N1 astrocytoma cell line expressing these markers increased significantly after treatment with TNFalpha or IL1beta. The presence of Abeta 1-40 fragment, alone or in combination with IL1beta, induced an increase in the percentage of cells expressing CD44, but not VCAM-1. However, the concomitant presence of Abeta 1-40 fragment and of IL1beta or TNFalpha caused an increase in the percentage of CD71 positive cells. In contrast, the shorter Abeta 25-35 fragment was always inactive. These results indicates that Abeta 1-40 fragment, in association with cytokines, can activate this astrocyte-derived cell line and add further elements in favour of the hypothesis that beta-amyloid can act as immunological mediator. PMID- 12767056 TI - Cdc42 and RhoA are differentially regulated during arachidonate-mediated HeLa cell adhesion. AB - Cell adhesion to extracellular matrix requires stimulation of an eicosanoid signaling pathway through the metabolism of arachidonate by 5-lipoxygenase to leukotrienes and cyclooxygenase-1/2 to prostaglandins, as well as activation of the small GTPase signaling pathway involving Cdc42 and Rho. These signaling pathways direct remodeling of the actin cytoskeleton during the adhesion process, specifically the polymerization of actin during cell spreading and the bundling of actin filaments when cells migrate. However, few studies linking these signaling pathways have been described in the literature. We have previously shown that HeLa cell adhesion to collagen requires oxidation of arachidonic acid (AA) by lipoxygenase for actin polymerization and cell spreading, and cyclooxygenase for bundling actin filaments during cell migration. We demonstrate that small GTPase activity is required for HeLa cell spreading upon gelatin, and that Cdc42 is activated while Rho is downregulated during the spreading process. Using constitutively active and dominant negative expression studies, we show that Cdc42 is required for HeLa cell spreading and migration, while activated RhoA is antagonistic towards spreading. Constitutively active RhoA promotes cell migration and increases the degree of actin bundling in HeLa cells. Further, we demonstrate that activation of either the AA oxidation pathway or the small GTPase pathway cannot rescue inhibition of spreading when the alternate pathway is blocked. Our results suggest (1) both the eicosanoid signaling pathway and small GTPase activation are required during HeLa cell adhesion, and (2) these signaling pathways converge to properly direct remodeling of the actin cytoskeleton during HeLa cell spreading and migration upon collagen. PMID- 12767058 TI - Nonsteroidal anti-inflammatory drug-activated gene (NAG-1) is induced by genistein through the expression of p53 in colorectal cancer cells. AB - Genistein is an isoflavenoid found in soy that has anti-tumorigenic activities. Treatment of colorectal carcinoma HCT-116 cells with 50 microM genistein results in a 50% reduction in cell proliferation and a 6-fold increase in apoptosis. Genistein induces nonsteroidal anti-inflammatory drug-activated gene 1 (NAG-1), a protein with antitumorigenic activities, in a time- and concentration-dependent manner in HCT-116 cells. In addition, p53 and p21 are induced in HCT-116 cells. The induction of p53 (3 hr) precedes the induction of NAG-1 (12 hr), suggesting that genistein-induced NAG-1 expression is mediated by p53. In contrast, NAG-1 is not induced by genistein in the p53-negative colorectal carcinoma cell line HCT 15. Luciferase reporter constructs of the NAG-1 promoter containing 2 p53 sites showed that the p53 sites within the NAG-1 promoter are critical to genistein induced NAG-1 expression in p53-positive U2OS cells. The expression of p53 was critical for NAG-1 promoter activity since no promoter activity was observed with genistein treatment in HCT-15 cells. However, genistein-induced promoter activity was restored in HCT-15 cells by transfection with wild-type p53. Together our data suggest a relationship between genistein, p53 and NAG-1 forming a novel pathway responsible for the antitumorigenic activity of genistein. PMID- 12767057 TI - Mitogenic and antiapoptotic role of constitutive NF-kappaB/Rel activity in pancreatic cancer. AB - The transcription factor NF-kappaB/Rel was found to be constitutively activated in human pancreatic cancer. RelA is present in the nucleus in primary human pancreatic cancer samples as well as in pancreatic cancer cell lines. NF kappaB/Rel-binding activity consists of NF-kappaB1(p50) and RelA(p65). Constitutive NF-kappaB/Rel activity correlates with IkappaB kinase (IKK) activity and can be blocked by dominant negative mutants of IKKbeta and to a lesser extent by IKKalpha. Constitutive NF-kappaB/Rel activity and the transactivation potential of RelA(p65) can be inhibited by dominant negative mutant Ras, the PI3 kinase inhibitor LY294002, or dominant negative mutant Akt kinase. Transfection of a dominant negative mutant epidermal growth factor receptor (EGF-R), EGF-R kinase inhibitor Tyrphostin and LY 294002 blocked IKK activity and NF-kappaB dependent transcription. Inhibition of constitutive IKK or NF-kappaB/Rel activity increased the number of apoptotic cells. Stably expressing a nondegradable form of IkappaBalpha inhibited anchorage-dependent and -independent proliferation in MiaPaCa2 and Panc1 cells. Our data demonstrate that an EGF-R/Ras/PI3 kinase/Akt/IKK-dependent pathway contributes to constitutive NF-kappaB/Rel activity in pancreatic cancer. Inhibition of NF-kappaB/Rel activity reveals a mitogenic and antiapoptotic role for NF-kappaB/Rel in pancreatic cancer. PMID- 12767059 TI - Expression of ADAM-9 mRNA and protein in human breast cancer. AB - The ADAMs (a disintegrin and metalloprotease) are membrane proteins containing both protease and adhesion domains and thus may be potentially important in cancer invasion and metastasis. The aim of our study was to investigate the distribution and potential clinical significance of ADAM-9 in breast cancer. ADAM 9 expression was measured using both reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting. ADAM-9 mRNA was expressed more frequently in both breast carcinomas (72/110, 66%) and fibroadenomas (21/38, 55%) compared to normal breast tissue (6/25, 24%) (p = 0.0004, p = 0.028, respectively). Multiple forms of ADAM-9 protein were detected by Western blotting, i.e., at 124, 84 and 48 kDa under reducing conditions and at 115, 76, 55, 52 and 46 kDa under nonreducing conditions. The 84 and 55 kDa forms were detected more frequently in the primary cancers compared to normal breast tissue (p < 0.0001, p = 0.0002, respectively). In addition, relative levels of the 84 kDa mature form were significantly higher in the primary cancers than in the fibroadenomas (p = 0.003), while the reverse was found for the 124 kDa precursor form (p = 0.026). In the carcinomas, the 84 kDa form of ADAM-9 protein was expressed at higher levels in node-positive than node-negative cancers (p = 0.05) and correlated positively with HER-2/neu protein levels (r = 0.313, p = 0.016). This is the first report to describe expression of any ADAM in a large number of human carcinomas. PMID- 12767061 TI - Transforming growth factor beta-1 and amphiregulin act in synergy to increase the production of urokinase-type plasminogen activator in transformed breast epithelial cells. AB - Amphireguline (AR) is an epidermal growth factor (EGF)-related peptide that seems to play an important role in breast cancer progression. We have demonstrated recently that suppression of AR expression in transformed breast epithelial cells considerably reduced both size and neovascularization of tumors developed in nude mice. We show that the reduction of AR expression allowed to an important decrease of the levels of urokinase-type plasminogen activator (uPA) and transforming growth factor-beta1 (TGFbeta1). According to these data, exogenous AR (10(-10) M-10(-8) M) stimulated the production of uPA and TGFbeta1 in AR antisense-transfected A2-15 and A2-P17F25 cells. The addition of 2 x 10(-10) M TGFbeta1 into culture medium increased the level of uPA produced by AR-expressing parental cells but not by A2-15 and A2-P17F25 cell clones. Whereas AR alone stimulated uPA production to 200% of control, combined AR and TGFbeta1 treatment increased protease level in A2-15 and A2-P17F25 cells to 500-600% of control, demonstrating a synergism between TGFbeta1 and AR. This was accompanied by an important augmentation of the number of tumoral cells that invaded matrigel in vitro. The synergistic induction of uPA protein resulted of an early and transient augmentation of steady state mRNA level and was blocked in the presence of the MAP kinase kinase inhibitor PD098059, strongly suggesting that synergistic effect of AR and TGFbeta1 on uPA expression required MAPK pathway. This data demonstrates concerted action between AR and TGFbeta1 that may have profound effect on protease production and consequently on breast cancer progression. PMID- 12767060 TI - Overexpression of RbAp46 facilitates stress-induced apoptosis and suppresses tumorigenicity of neoplastigenic breast epithelial cells. AB - We have found previously that the retinoblastoma (Rb) suppressor associated protein 46 (RbAp46) is a gene upregulated by the Wilms' tumor suppressor, WT1, and functions as a potent growth inhibitor. To investigate the effect of RbAp46 overexpression on early development of breast cancer, we established stable cell lines from neoplastigenic breast epithelial cells, MCF10AT3B, a cell line derived from a model of human proliferative disease, to constitutively express exogenous RbAp46. We have found that expression of RbAp46 suppressed colony formation of MCF10AT3B cells in soft-agar, and inhibited tumor formation of these cells in nude mice. Expression of RbAp46 sensitized MCF10AT3B cells to apoptosis induced by serum deprivation and hydrocortisone withdrawal. Furthermore, we have found that the c-Jun NH2-terminal kinase (JNK) pathway and GADD45, a growth arrest- and DNA damage-inducible gene, are constitutively activated in RbAp46-expressing cells. Our data suggested that high levels of RbAp46 expression inhibit the tumorigenicity of neoplastigenic breast epithelial cells by facilitating JNK dependent apoptotic cell death. Our data also suggested that dysregulation of RbAp46 gene may be involved in the early development of breast cancer. PMID- 12767062 TI - Increased adhesion potency of ovarian carcinoma cells to mesothelial cells by overexpression of dipeptidyl peptidase IV. AB - Dipeptidyl peptidase IV (DPPIV), cell surface peptidase, works as an adhesion molecule as well as an enzyme. To investigate the role of DPPIV as an adhesion molecule of ovarian carcinoma cells to mesothelial cells, ovarian carcinoma (SKOV3 and NOS4) cells were transfected with a sense and an antisense cDNA coding human DPPIV. The adhesion potency of these transfected cells to fibronectin- and collagen-coated plates and mesothelial cells were examined with or without fibronectin. Flow cytometry and immunohistochemistry demonstrated DPPIV in ovarian carcinoma cells. The adhesion rate of DPPIV-transfected SKOV3 (SKDPIV) cells to fibronectin-coated plates was significantly higher than SKOV3 cells, while there was no difference in the adhesion rate to non-coated plates between SKDPIV and SKOV3 cells. The adhesion rates of vector-transfected SKOV3 (SKpcDNA) cells to coating and noncoating plates were similar to those of SKOV3 cells. SKDPIV cells showed twofold adhesion potency to mesothelial cells compared to SKOV3 cells. Furthermore, an addition of soluble fibronectin resulted in a dose dependent increase in the adhesion rate of SKDPPIV cells, but not in either SKOV3 or SKpcDNA cells. Antisense-transfected NOS4 cells decreased the adhesion potency compared to NOS4 and vector-transfected NOS4 cells. In conclusion, ovarian carcinoma cells express DPPIV and adhere to human mesothelial cells in part by DPPIV. This adhesion mechanism of DPPIV is mediated by immobilized and soluble fibronectin. PMID- 12767063 TI - Novel plant triterpenoid drug amooranin overcomes multidrug resistance in human leukemia and colon carcinoma cell lines. AB - Amooranin (AMR), a plant terpenoid, isolated from Amoora rohituka, was investigated for its ability to overcome multidrug resistance in human leukemia and colon carcinoma cell lines. AMR IC(50) values of multidrug-resistant leukemia (CEM/VLB) and colon carcinoma (SW620/Ad-300) cell lines were higher (1.9- and 6 fold) than parental sensitive cell lines (CEM and SW620). AMR induced G(2)+M phase-arrest during cell cycle traverse in leukemia and colon carcinoma cell lines and the percentage of cells in G(2)+M phase increased in a dose-dependent manner. Coincubation of tumor cells with both DOX and AMR reversed DOX resistance in 104-fold DOX-resistant CEM/VLB and 111-fold DOX-resistant SW620/Ad-300 cell lines with a dose modification factor of 50.9 and 99.6, respectively. Flow cytometric assay showed that AMR causes enhanced cellular DOX accumulation in a dose-dependent manner. AMR inhibits photolabeling of P-glycoprotein (P-gp) with [(3)H]-azidopine and the blocking effect enhanced with increasing concentrations of AMR. Our results show that AMR competitively inhibits P-gp-mediated DOX efflux, suggestive of a mechanism underlying the enhanced DOX accumulation and reversal of multidrug resistance by AMR. PMID- 12767064 TI - Upregulation of MMPs by soluble E-cadherin in human lung tumor cells. AB - Loss of E-cadherin/catenin mediated cell-cell adhesion and overexpression of matrix metalloproteinases (MMPs) are largely involved in tumor invasion. It has been recently shown that high levels of a soluble 80 kDa fragment of E-cadherin, resulting from a cleavage by MMPs, are found in serum and in urine from cancer patients. Additionally, this soluble E-cadherin (sE-CAD) promotes cell invasion into chick heart and into collagen type I gels. The aim of our study was to examine the mechanism of sE-CAD-induced cell invasion. Since MMPs play a crucial role in invasion, we looked for induction of MMPs by sE-CAD in noninvasive human lung tumor cells 16HBE. An induction of MMP-2, MMP-9 and MT1-MMP expression was observed both at the mRNA and at the protein level in the presence of sE-CAD (in conditioned medium form or in E-cadherin HAV peptide form). No induction of MMP 1, -3 and -7 or variation of the levels of their inhibitors, TIMP-1 and TIMP-2, were detected. The biologic relevance of the sE-CAD-induced MMP upregulation was tested by demonstrating that sE-CAD promotes in vitro cell invasion in a modified Boyden chamber assay. These data provide new insight into mechanisms of tumor invasion by ectodomain shedding of the cell-cell adhesion molecule E-cadherin. PMID- 12767065 TI - HER 2/neu expression and gene amplification in colon cancer. AB - HER 2/neu is an important oncogene in breast cancer, but the prevalence and significance of HER 2/neu gene amplification in colon cancer have been poorly documented. We have evaluated HER 2/neu gene amplification and protein overexpression in a series of colon cancers to assess the frequency, concordance and clinical significance of these events. HER 2/neu gene copy number was measured in 154 primary colon tumors, 15 liver metastases and matched normal tissues using a quantitative PCR/ligase detection reaction (LDR) technique developed and validated in our laboratory. HER 2/neu copy number was confirmed by fluorescent in situ hybridization (FISH) in all tumors found to have gene amplification. In an independent and blinded fashion, HER 2/neu expression was assessed in paraffin sections from 139 of the tumor specimens using the HercepTest kit. HER 2/neu gene amplification was observed in 4 (2.4%) of the 169 tumor specimens and in none of the normal tissues. There was no apparent association with stage of disease, tumor grade or patient survival. Among 139 cases evaluated by immunohistochemistry (IHC), HER 2/neu overexpression was seen in 5 cases (3.6%). There was extremely high concordance (kappa = 0.852) between gene amplification and protein overexpression. The low prevalence of HER 2/neu gene amplification and protein overexpression suggests that this oncogene plays an infrequent role in the development and progression of colon cancer. These data indicate that the primary mechanism of dysregulated HER 2/neu expression in colon cancer, as in breast cancer, is gene amplification. PMID- 12767066 TI - Downregulation of Cap43 gene by von Hippel-Lindau tumor suppressor protein in human renal cancer cells. AB - We previously identified 9 genes (i.e., thymosin beta4, secreted protein acidic and rich in cysteine, Cap43, ceruloplasmin, serum amyloid A, heat shock protein 90, LOT1, osteopontin and casein kinase Igamma) that are more highly expressed in cancerous regions than in noncancerous regions in human renal cancers. In our study, we considered the possibility that the von Hippel-Lindau (VHL) tumor suppressor gene might be able to affect the expression of these 9 genes in renal cancer cells. We first established 2 VHL-positive cell lines, 786/VHL-1 and 786/VHL-2, after the introduction of wild-type VHL into VHL-negative renal cancer 786-O cells. Of these 9 genes, expression of the Cap43 gene was specifically downregulated by VHL. Expression of Cap43 was also much lower in 4 other VHL positive renal cancer cell lines than in VHL-negative 786-O cells. Cap43 promoter assays with several deletion or mutation constructs demonstrated that the Sp1 site in the element from -286 base pairs (bp) to -62 bp was partly responsible for VHL-induced suppression of the Cap43 gene. Immunostaining analysis with human specimens of renal cancers demonstrated that the Cap43 protein was expressed in most cancer cells and macrophages. We also observed a marked and specific increase of Cap43 mRNA levels in response to hypoxia or nickel in all VHL positive cell lines. Cellular expression of Cap43 mRNA in response to hypoxia or nickel thus is closely associated with VHL gene expression in renal cancer cells. Although the function of the Cap43 protein remains unclear, the expression of Cap43 protein could be a molecular marker closely associated with VHL in renal cancer. PMID- 12767067 TI - Recombinant E. coli efficiently delivers antigen and maturation signals to human dendritic cells: presentation of MART1 to CD8+ T cells. AB - The generation of tumour-specific cytotoxic T-lymphocyte (CTL) responses is the primary focus in the design of immunotherapeutic cancer vaccines. We have recently demonstrated generation of ovalbumin (OVA)-specific CTLs and tumour protection in a murine tumour model using vaccination with dendritic cells (DCs) pulsed with E. coli expressing listeriolysin O (LLO) and OVA as a model antigen. In this system paraformaldehyde fixation of E. coli/LLO provided an additional safety feature without compromising vaccine efficacy. We therefore reasoned that paraformaldehyde-fixed recombinant E. coli expressing LLO would be an efficient vehicle for the delivery of human tumour antigens to human DCs. In the present study, we demonstrate that fixed E. coli expressing LLO are taken up efficiently by human monocyte-derived DCs (MoDCs) with minimal toxicity. As a consequence of the interaction with bacteria, human DCs undergo marked phenotypic and functional maturation. Furthermore, we show that fixed E. coli/LLO expressing the well characterised human melanoma antigen, MART1, efficiently deliver the HLA-A2 restricted MART1(27-35) epitope for processing and presentation on human MoDCs, suggesting the potential of this system as a novel strategy for human tumour immunotherapy. PMID- 12767068 TI - Induction of systemic immunity by expression of interleukin-23 in murine colon carcinoma cells. AB - Interleukin-23 (IL-23), a novel cytokine composed of a newly identified p19 molecule and the p40 subunit of IL-12, can stimulate the proliferation in vitro of memory T cells. We examined whether Colon 26 murine colon carcinoma cells that were retrovirally transduced with the p19-linked p40 gene (Colon 26/IL-23) could produce antitumor effects in inoculated mice. The growth of Colon 26/IL-23 tumors developed in immunocompetent mice was significantly retarded and the tumors disappeared thereafter. Spleen cells from the mice that received Colon 26/IL-23 cells produced significant amounts of interferon-gamma, when they were cultured with irradiated Colon 26 but not irrelevant cells. Depletion of CD8(+) T cells suppressed the production of interferon-gamma. The mice that had rejected Colon 26/IL-23 tumors were resistant to subsequent challenge of parent but not irrelevant tumor cells. Colon 26/IL-23 tumors were not rejected in nude mice but the growth was retarded compared to parent tumors. Treatment of nude mice with anti-asialo GM(1) antibody did not influence the growth of Colon 26/IL-23 tumors. These data suggest that expression of IL-23 in tumors produces T cell-dependent antitumor effects and induces systemic immunity. PMID- 12767069 TI - A national case-control study of Ewing's sarcoma family of tumours in Australia. AB - Limited population-based epidemiologic information is available on Ewing's sarcoma family of tumours (ESFT), a rare group of neoplasms. Several associations have been noted on a few studies but results were not consistent, except for exposure to farming among cases and their parents. Here we present the non-farm findings of a nationwide case-control study of ESFT in children and young adults in Australia. The analysis included 106 persons with confirmed ESFT and 344 population-based controls selected randomly via telephone. Information was collected by interview (84% face to face). We found a strong and significant association of ESFT with hernias, in particular hernia repaired in hospital (OR = 5.6, 95% CI 1.3-6.4). Among other factors, there was a near doubling of risk for males, and male cases had their pubertal signs earlier (started shaving earlier) than male controls. There was also an increased risk of ESFT at higher levels of self-assessed exercise, but no other factor really stood out. For pregnancy related factors, there was a tripling of risk for glandular fever, a doubling of risk for urinary tract infection and a near doubling of risk for X-rays during or just before pregnancy, but these estimates were not significant. In addition, there was a large number of inverse associations with medical conditions (specifically bone disorders), case exposure to medications, vaccinations and X rays, with ultrasound during the pregnancy having the most certain effects. We conclude that, although the aetiology of ESFT remains obscure, overall there is strong evidence of an association with inguinal hernia; this can now be added to the farm-associated risk reported by others and us. The other associations reported here await replication and refinement in future studies. PMID- 12767070 TI - Physical inactivity, energy intake, obesity and the risk of rectal cancer in Canada. AB - We conducted a population-based case-control study of 1,447 incident rectal cancer cases and 3,106 population controls aged 20-76 years to assess the effect of recreational physical activity, energy intake and obesity on rectal cancer risk in 7 of 10 Canadian provinces in 1994-97. After adjustment for the effect of various potential confounding factors, total recreational physical activity in the highest quartile was associated with an odds ratio (OR) for rectal cancer risk of 0.88 (95% confidence interval [CI] = 0.64-1.20) in women and 1.15 (95% CI = 0.88-1.49) in men. Women and men in the highest quartile of caloric intake (> = 56,741 and > = 63,143 kJ/week) had ORs of 1.50 (95% CI = 1.00-2.25) and 1.61 (95% CI = 1.13-2.28), respectively. Total dietary fat intake was not associated with a risk of rectal cancer after adjustment for caloric intake. Obesity (BMI > = 30 kg/m(2)) was associated with an OR of 1.44 (95% CI = 1.06-1.95) for women and 1.78 (95% CI = 1.36-2.34) for men. Men and women with lifetime maximum body mass index (BMI) > = 30 kg/m(2) had respective ORs of 1.70 (95% CI = 1.30-2.23) and 1.26 (95% CI = 0.96-1.66). The greatest increase in rectal cancer risk was observed in men and women with simultaneous high energy intake, high BMI and low physical activity. Our study provides evidence that physical inactivity, high energy intake and obesity are associated with the risk of rectal cancer, and there is a probable synergic effect among the 3 risk factors. PMID- 12767071 TI - Physical activity and epithelial ovarian cancer risk: a case-control study in China. AB - A case-control study was conducted in China during 1999-2000 to investigate the effects of intensity and duration of physical activity on the risk of epithelial ovarian cancer. Cases were 254 patients with histologically confirmed epithelial ovarian cancer. The 652 controls comprised 340 hospital visitors, 261 non neoplasm hospital outpatients and 51 women recruited from the community. Physical activity was measured by a validated questionnaire. The risks of ovarian cancer were assessed using multivariate logistic regression analysis accounting for age, demographic, lifestyle and familial factors, hormonal status, family ovarian cancer history and total energy intake. The study found that increasing total physical activity was associated with a lower ovarian cancer risk among Chinese women. The odds ratio was 0.54 (95% CI 0.34-0.87) for high vs. low levels of total weekly metabolic equivalent tasks. Ovarian cancer risk tended to decline with increasing duration of strenuous sports and frequency of activity-induced sweating among pre-menopausal women, with adjusted OR 0.13 (95% CI 0.03-0.64) and 0.45 (95% CI 0.24-0.85), respectively. Increasing duration of moderate activity in post-menopausal women also appeared to be protective against ovarian cancer, with adjusted OR 0.36 (95% CI 0.18-0.73). PMID- 12767072 TI - Breast cancer and specific types of oral contraceptives: a large Norwegian cohort study. AB - The aim of our study was to examine the risk of breast cancer according to specific types of estrogens and progestagens in oral contraceptives (OCs) based on the prospective Norwegian Women and Cancer study (NOWAC). Between 1991-97 women aged 30-70 years were drawn at random from the central person register and mailed an invitation and a questionnaire. Women (102,443) were enrolled with follow-up information collected throughout 1999 by linkage with national registries of cancer, mortality and emigration based on the unique national identification number. Among the 96,362 women included in the present analysis 851 invasive breast cancer were diagnosed. The adjusted risk of breast cancer increased with 25% for ever use of OCs and the risk increased with increasing duration of use (test for trend: p = 0.007). No association between time since last use and breast cancer risk was found after stratification on duration of use. Positive trend was still found for total duration of use among women who used OCs more than 5 years ago. Second generation of OCs had an increased risk with increasing duration of use. Classifying progestagens according to chemical groups, the relative risk increased significantly with increasing cumulative dose of levonorgestrel progestagen. It was difficult to conclude for the other groups due to lack of power. In a multivariate analysis the cumulative dose for all progestagen groups were non-significant, although we observed a significant increased risk with increasing milligram-months of estrogen exposure (p = 0.002). In conclusion, the increased risk of breast cancer related with OC formulations could be due mostly to estrogen component. PMID- 12767073 TI - Evaluation of hypermethylated tumor suppressor genes as tumor markers in mouth and throat rinsing fluid, nasopharyngeal swab and peripheral blood of nasopharygeal carcinoma patient. AB - The purpose of our study was to evaluate the frequency of hypermethylated tumor suppressor genes (TSGs) in peripheral blood, mouth and throat (M&T) rinsing fluid and nasopharyngeal (NP) swabs of nasopharyngeal carcinoma (NPC) patients. Six normal NP tissues, 43 M&T rinsing fluid, 37 NP swabs and 43 peripheral blood from healthy non-smokers and non-drinkers without a family history of NPC, and 30 NPC tumors and their matched body fluid were analyzed for the presence of hypermethylated p15, p16, Ras association domain family 1 (RASSF1A), E-cadherin, and death-associated protein kinase (DAPK) by methylation-specific PCR. Sequencing analysis was carried out on selected NPC tumors and body fluid samples. Twenty-nine (97%) tumors displayed methylation in at least 1 of the 5 genes. The methylation frequencies were 80% for p15, 77% for DAPK, 67% for RASSF1A, 53% for E-cadherin and 33% for p16. The frequency range of aberrant methylated genes in the body fluids were NP swabs (17-63%) and M&T rinsing fluid (17-50%). Methylation was found in <20% of peripheral blood for each respective gene. Methylation was, however, detected in 1 M&T rinsing fluid in which the primary tumor showed methylation free for RASSF1A. Five healthy individuals exhibited methylation for DAPK, or RASSF1A, or p15 in their body fluid samples. All body fluid samples of healthy controls showed methylation free for E-cadherin and p16. Epigenetic change is found frequently in NPC and the high detection rate in body fluids suggest its potential application in non-invasive screening of NPC or detection of residual carcinoma after treatment. PMID- 12767074 TI - Growth inhibitory retinoid effects after recruitment of retinoid X receptor beta to the retinoic acid receptor beta promoter. AB - Nuclear retinoid receptors mediate retinoid effects through tissue-specific, ligand-receptor interactions and subsequent transcriptional regulation of secondary target genes. Retinoic acid receptor beta (RARbeta) is itself a retinoid target gene with a retinoic acid response element (betaRARE) in the 5' untranslated region of the RARbeta2 gene. Altered transcriptional regulation of RARbeta may play a role in human carcinogenesis and the retinoid-responsiveness of malignant cells. Here we used retinoid X receptor-specific antibodies in electrophoretic mobility shift assays to show that the retinoid X receptor beta (RXRbeta) protein was recruited to the betaRARE, after retinoid treatment of retinoid-sensitive neuroblastoma (NB), lung and breast cancer cell lines, but not retinoid-resistant lung and breast cancer cell lines. RXRbeta selectively enhanced retinoid-induced transcriptional activation of the betaRARE. Stable overexpression of RXRalpha and RXRbeta in NB cells resulted in marked growth inhibition and cell death, which increased after retinoid treatment. However, only proteins from the RXRbeta transfectants exhibited specific RXRbeta binding to the betaRARE in vitro and in vivo, enhanced histone acetylation and increased endogenous RARbeta expression. These data indicate that recruitment of RXRbeta to the betaRARE, and consequent induction of endogenous RARbeta expression, is an important component in the retinoid anticancer signal. RXRalpha may also participate in the retinoid signal, but through mechanisms that do not involve RARbeta. PMID- 12767076 TI - Biodistribution and pharmacokinetics of 125I-labeled monoclonal antibody M75 specific for carbonic anhydrase IX, an intrinsic marker of hypoxia, in nude mice xenografted with human colorectal carcinoma. AB - Carbonic anhydrase IX (CA IX) is frequently expressed in human carcinomas and absent from the corresponding normal tissues. Strong induction by tumor hypoxia predisposes CA IX to serve as a target for cancer diagnostics and therapy. Here we evaluated targeting properties and pharmacokinetics of CA IX-specific monoclonal antibody (MAb) M75. Binding parameters of (125)I-labeled M75, including equilibrium dissociation constant, hypoxia-related binding to various cell lines and internalization, were analyzed in vitro. Biodistribution of (125)I M75 in nude mice bearing HT-29 human colorectal carcinoma xenografts with hypoxic pattern of CA IX expression was studied by measurements of radioactivity in dissected tissues and macroautoradiography of tissue sections. Pharmacokinetics of intravenously administered (125)I-M75 was described using a 2-compartment model. Blood clearance showed a distribution phase t(1/2)(alpha) = 3.4 hr and an elimination phase t(1/2)(beta) = 55.3 hr postinjection. Despite predominant CA IX localization in less accessible perinecrotic regions, (125)I-M75 exhibited specific accumulation in xenograft, with a mean uptake of 15.3 +/- 3.6% of injected dose per gram of tumor tissue at 48 hr postadministration. Specificity of M75 localization was confirmed by low tumor uptake of control antibody. Altogether, our data demonstrate that M75 MAb is a promising tool for selective immunotargeting of hypoxic human tumors that express CA IX. PMID- 12767075 TI - Increased toxicity and lack of efficacy of Rofecoxib in combination with chemotherapy for treatment of metastatic colorectal cancer: A phase II study. AB - Preclinical and clinical models have demonstrated that cyclooxygenase-2 (COX-2) is overexpressed in primary and metastatic colorectal tumors. In preclinical models, there appears to be additive or synergistic effects when combining 5 Fluorouracil (5-FU) with nonsteroidal anti-inflammatory agents (NSAIDs) for the treatment of colorectal neoplasms. This data raised the question as to whether adding a COX-2 inhibitor to 5-FU-based regimens would increase the response rates with an acceptable toxicity profile in patients with metastatic colon cancer. In the current study, patients with metastatic colorectal cancer, who were either untreated or previously treated (more than 1 year ago) with adjuvant 5-FU and Leucovorin (LV) received 5-FU and LV (Mayo regimen) in addition to Rofecoxib. Tumor samples from all patients exhibited evidence of moderate COX-2 over expression. 4 patients entered on the study developed upper gastrointestinal bleeding (grade III). Other toxicities included grade II stomatitis (3 patients), grade II thrombocytopenia (1 patient), grade II diarrhea (2 patients) and grade I nausea (1 patient). There were no partial or complete responses in the first 10 patients entered on the study so the study was terminated (probability of success < 0.3 with type 1 error of 0.05 and power of 0.8). Thus, Rofecoxib did not appear to increase antitumor activity and resulted in increased gastrointestinal toxicity when combined with 5-FU/LV. Future studies will need to consider the added gastrointestinal toxicity of Rofecoxib when combined with chemotherapy for the treatment of patients with colorectal cancer. PMID- 12767081 TI - Reflections of a cancer survivor/research scientist. AB - For more than 50 years, Cancer has been devoted to exploring every facet of malignant neoplasia, from morphology to therapy, from epidemiology to biology, from historical perspective to basic science. One aspect, however, needs emphasis, namely, the human being who bears the burden of the cancer. For this reason and with the encouragement of the Editor, we have undertaken to remedy this situation by directing attention to the fears and frustrations, hopes and expectations, and feelings and thoughts of all kinds of the person who has cancer, who is worried about it, and who is dependent on physicians specialized in its diagnosis and management. For doctors to be understanding of the very deep needs of patients with cancer, it is requisite that they be privy to authentic, but often unspoken, expressions of a patient's anxieties about physical and emotional pain, loss of control over personal destiny, and plain dread of dying and of the end of cherished relationships. PMID- 12767082 TI - Matrix metalloproteinase inhibitor reversion-inducing cysteine-rich protein with Kazal motifs: a prognostic marker for good clinical outcome in human breast carcinoma. AB - BACKGROUND: The recently described reversion-inducing cysteine-rich protein with Kazal motifs (RECK) inhibits membrane Type 1 matrix metalloproteinase (MMP-14), MMP-2, and MMP-9 secretion and enzymatic activity. Its expression is essential for normal vasculogenesis. Down-regulation of RECK has been implicated in tumor angiogenesis and progression. METHODS: The authors assessed the prognostic value of RECK expression in tumor tissue specimens from 278 breast carcinoma patients with a median follow-up time of 75 months (range, 2-169 months). RECK mRNA levels were measured by real-time quantitative reverse transcriptase-polymerase chain reaction. RESULTS: Expression levels of RECK were lower in tumor tissue specimens than in adjacent normal breast tissue specimens from 10 patients (P = 0.028). No relevant associations of RECK with established clinicopathologic factors or treatment regimens were found. RECK expression predicted a longer recurrence-free survival time (RFS; P = 0.037) at the optimal cutoff value (hazard ratio, 0.66; 95% confidence interval, 0.44-0.98). The 100 patients whose tumors exhibited low levels of RECK had a mean RFS time of 80.4 months and a 61.8% 5-year RFS rate, whereas the 178 patients with tumors with high RECK expression had a mean RFS time of 91.2 months and a 73.0% 5-year RFS rate. Multivariate Cox regression analysis showed that RECK expression maintained a significant independent prognostic value for RFS time (P = 0.047). CONCLUSIONS: These results are in agreement with the notion of RECK being an important tumor-suppressor gene. Therefore, the possibility of applying RECK, a pharmaceutical mimetic, or drugs activating endogenous RECK expression, as possible therapeutic or preventive agents for breast carcinoma should be explored. PMID- 12767083 TI - A phase III trial of sequential adjuvant chemotherapy for operable breast carcinoma: final analysis with 10-year follow-up. AB - BACKGROUND: The current study was performed to assess whether sequential potentially noncross-resistant chemotherapy prolongs disease-free survival (DFS) and overall survival (OS) in patients with operable breast carcinoma. METHODS: Seven hundred eighty-nine patients were registered and followed for a median of 10 years. They were treated in two groups. In Group 1, patients age < 50 years or age > 50 years but with either negative or unknown estrogen receptor (ER) status were randomized to receive 6 cycles of 5-fluorouracil, doxorubicin, and cyclophosphamide (FAC) alone or followed by 4 cycles of methotrexate and vinblastine (MV). In Group 2, patients age > or = 50 years with ER-positive disease were randomized to receive either tamoxifen or combination chemotherapy (FAC + MV) for 10 cycles. Analysis was performed according to allocated treatment (intention to treat), with all randomized patients included. RESULTS: In Group 1 there were no significant differences with regard to DFS or OS between the two treatment arms. The DFS at 5 years was 0.70 and 0.76, respectively, for FAC compared with FAC+MV (P = 0.26). The OS was similar for both groups (0.84 vs. 0.83). It is interesting to note that there was a statistically nonsignificant trend for improved DFS in the FAC + MV arm for patients who were ER-positive. In Group 2, tamoxifen alone led to more prolonged DFS compared to FAC+MV (0.78 vs. 0.66, respectively) but this did not reach statistical significance (P = 0.28). OS also was associated with a trend (P = 0.86) toward prolonged survival for the tamoxifen arm compared with the FAC+MV arm (0.85 vs. 0.74, respectively). CONCLUSIONS: The results of the current trial concerning sequential adjuvant chemotherapy for operable breast carcinoma, which to our knowledge contains the longest follow-up presented to date, failed to demonstrate any additional benefit from the addition of 4 cycles of MV to 6 cycles of FAC chemotherapy. PMID- 12767084 TI - Tailored messages, external barriers, and women's utilization of professional breast cancer screening over time. AB - BACKGROUND: The efficacy of tailored messages on women's utilization of professional breast screening over time has not been examined often or in combination with external barriers (e.g., costs and access). The objectives of this study were to test the effects of alternative messages on mammography and clinical breast examination (CBE) utilization over time and to examine the combined effects of such messages and external barriers. METHODS: In a randomized, controlled trial, three message conditions were compared: no message (control group), recommendations about screening, or recommendations plus tailored discussion (on beliefs, feelings, costs, and access). Messages were delivered by advanced practice nurses over the telephone. Participants were women ages 51-80 years who had not had mammography in the prior 13 months. Outcome measures were women's mammography and CBE utilization 3-6 months postintervention (short-term follow-up) and 13-16 months after short-term follow-up (long-term follow-up). RESULTS: Both messages promoted mammography and CBE utilization at short-term follow-up. Utilization increased over time in all groups. Mammography utilization was greater for the tailored-message group compared with the recommendations-only group at long-term follow-up. Messages and external barriers had combined effects (Ps < 0.01). Among participants with high external barriers, participants in the message conditions-especially the tailored message-had the highest screening rates; among participants with low barriers, screening rates were similar across conditions. CONCLUSIONS: Clinicians can tailor discussions on beliefs, feelings, cost, and access about screening to promote professional breast screening over time, especially with women who have not had mammograms as recommended and who have external barriers to screening. PMID- 12767085 TI - Long-term results in elderly patients with differentiated thyroid carcinoma. AB - BACKGROUND: The authors assessed the biologic behavior of differentiated thyroid carcinoma in patients age 70 years or older and evaluated factors that influenced long-term survival. METHODS: Among 1448 patients with differentiated thyroid carcinoma who were treated at the authors' institution over the past 60 years, 111 patients were identified who were age 70 years or older at the time of their initial diagnosis (range, 70-93 years). The authors conducted a retrospective analysis of the outcome of these 111 patients, who had a median follow-up of 9 years (range, 2-9 years). RESULTS: There were 83 female patients and 28 male patients (female to male ratio, 3:1). Fifty-eight tumors were papillary, 46 tumors were follicular, and 7 tumors were Hurthle cell carcinoma. Eighty percent of patients presented with a thyroid mass, and 70% of tumors were pathologic stage T3 (pT3) or pT4. Lymph node disease was evident at presentation in 44% of patients, and distant metastases were documented at diagnosis in 23% of patients. Forty-six patients underwent total thyroidectomy, and the remaining patients underwent subtotal thyroidectomy or biopsy only. Radioiodine was administered to 80 patients, and external beam radiotherapy was administered to 19 patients. The cause specific survival rates were 75%, 50%, and 50% at 5 years, 10 years, and 15 years, respectively. Multivariate analysis showed that the presence of metastases was the most important independent prognostic factor for survival. External beam radiotherapy was associated with a poorer prognosis, in that it was reserved for patients with either inoperable disease or residual disease after surgery and patients with no uptake of radioiodine. CONCLUSIONS: A large proportion of thyroid tumors showed extrathyroid spread and distant metastases, which frequently were not iodine-avid. The prevalent histologic type was papillary, often with features of poor differentiation. Thyroid carcinoma in the elderly appears to behave more aggressively, and they have a less favorable prognosis compared with younger adults. PMID- 12767086 TI - Phase II trial of gemcitabine and docetaxel in patients with advanced carcinoma of the urothelium: a trial of the Eastern Cooperative Oncology Group. AB - BACKGROUND: Gemcitabine and docetaxel are active agents in advanced urothelial carcinoma. A Phase II trial of this combination was performed to determine the activity and toxicity of these agents in a multiinstitutional setting in patients previously treated with one prior chemotherapy regimen. METHODS: Twenty-nine eligible patients with advanced urothelial carcinoma were treated with docetaxel at a dose of 40 mg/m(2) over 1 hour followed by gemcitabine, 800 mg/m(2), over 30 minutes, both intravenously (i.v.) on Days 1 and 8. Cycles were repeated every 21 days until disease progression or a maximum of 6 cycles. RESULTS: Five patients obtained an objective response for an overall response rate of 17% (90% confidence interval, 7-33%). One patient achieved a complete clinical response. The median overall survival of the group was 7.7 months. Toxicity was moderate with granulocytopenia, anorexia, and fatigue being the most commonly noted side effects. CONCLUSIONS: Gemcitabine and docetaxel is an active second-line combination in patients with advanced urothelial carcinoma. Responses in visceral, lymph node, and soft tissues sites were observed. Granulocytopenia without fever, fatigue, and anorexia was common. Thromboembolic symptoms were reported and are of concern. The combination of gemcitabine and docetaxel has the potential to palliate a subset of previously treated patients with an adequate performance status. PMID- 12767087 TI - High-dose sequential chemotherapy and peripheral blood progenitor cell autografting in patients with refractory and/or recurrent Hodgkin lymphoma: a multicenter study of the intergruppo Italiano Linfomi showing prolonged disease free survival in patients treated at first recurrence. AB - BACKGROUND: The objective of the current study was to evaluate in a multicenter setting the feasibility and efficacy of a high-dose sequential (HDS) chemotherapy regimen that combined intensive debulking and high-dose therapy (HDT) with peripheral blood progenitor cell (PBPC) autografting in patients with refractory or recurrent Hodgkin lymphoma (HL). METHODS: Data were collected from 102 patients with HL who were treated with the HDS regimen at 14 centers associated with the Intergruppo Italiano Linfomi. Twenty-four patients had primary refractory HL, 59 patients had their first recurrence of HL (within 1 year in 32 patients and > 1 year in 27 patients), and 19 patients had multiple disease recurrences. The HDS regimen included the sequential delivery of high-dose (hd) cyclophosphamide with PBPC harvesting, methotrexate, etoposide, then HDT (usually hd mitoxantrone plus L-phenylalanine mustard) with PBPC autografting. In addition, radiotherapy was delivered to 36 patients at sites of bulky or persistent disease. RESULTS: Ninety-two patients (90%) completed the HDS program. There were five toxic deaths (treatment-related mortality rate, 4.9%) and six secondary malignan cies (five patients developed myelodysplastic syndrome/acute myelogenous leukemia, and one patient developed colorectal carcinoma). At a median follow-up of 5 years, the 5-year overall survival (OS) and event-free survival (EFS) projections were 64% (95% confidence interval [95% CI], 54-74%) and 53% (95% CI, 43-63%), respectively. Patients with their first recurrence had the most favorable outcome, with 5-year OS and EFS projections of 77% (95% CI, 66 88%) and 63% (95% CI, 50-76%), respectively. There were no significant differences between patients with early first recurrence and late first recurrence. The poorest outcome was observed in patients with refractory HL, with 5-year OS and EFS projections of 36% (95% CI, 16-55%) and 33% (95% CI, 14-52%), respectively. Patients who received HDS chemotherapy after multiple recurrences had an intermediate outcome. Multivariate analysis showed that refractory disease and systemic symptoms at the time of initial presentation were associated significantly associated with poor OS and EFS. CONCLUSIONS: The use of HDS chemotherapy for patients with refractory and/or recurrent HL is feasible at the multicenter level. The combination of intensive debulking and HDT with PBPC autografting offers a good chance of prolonged disease free survival for patients with their first recurrence of HL. PMID- 12767088 TI - Results of imatinib mesylate therapy in patients with refractory or recurrent acute myeloid leukemia, high-risk myelodysplastic syndrome, and myeloproliferative disorders. AB - BACKGROUND: Imatinib mesylate is a selective tyrosine kinase inhibitor of c-abl, bcr/abl, c-kit, and platelet-derived growth factor-receptor (PDGF-R). c-kit is expressed in most patients with acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS) and PDGF has been implicated in the pathogenesis of myeloproliferative disorders (MPD). METHODS: The authors investigated the efficacy of imatinib in patients with these disorders. Forty-eight patients with AML (n = 10), MDS (n = 8), myelofibrosis (n = 18), atypical chronic myeloid leukemia (CML; n = 7), chronic myelomonocytic leukemia (CMML; n = 3), or polycythemia vera (n = 2) were treated with imatinib 400 mg daily. RESULTS: None of the patients with AML or MDS responded. Among patients with myelofibrosis, 10 of 14 patients with splenomegaly (71%) had a 30% or greater reduction in spleen size, 1 patient had trilineage hematologic improvement, 2 had erythroid hematologic improvement, and 1 had improvement in platelet count. One patient with atypical CML had erythroid hematologic improvement. Both patients with polycythemia vera needed fewer phlebotomies (from 2-3 per year to none during the 8 months of therapy and from 3-6 per year to 1 during 9 months of therapy). None of the three patients with CMML responded. Treatment was well tolerated. The side effects were similar to those observed in patients with CML. CONCLUSIONS: Within these small subgroups of disease types, single-agent imatinib did not achieve a significant clinical response among patients with AML, MDS, atypical CML, or CMML without PDGF-R fusion genes. Preliminary data on polycythemia vera are promising and deserve further investigation. Responses among myelofibrosis patients were minor. Therefore, a combination treatment regimen including imatinib may be more effective. PMID- 12767089 TI - Elevated serum endostatin is associated with poor outcome in patients with non Hodgkin lymphoma. AB - BACKGROUND: Endostatin is a cleaved fragment of collagen Type XVIII and has antiangiogenic activity. The clinical significance of circulating, soluble endostatin (S-endostatin) is not known. METHODS: Pretreatment S-endostatin and serum vascular endothelial growth factor (S-VEGF) levels were measured in 143 patients with non-Hodgkin lymphoma (NHL) using competitive enzyme immunoassays and were compared with the levels from a control group (n = 24 participants). RESULTS: S-endostatin levels varied widely from 4.5 ng/mL to 116 ng/mL (median, 29.6 ng/mL), and the median level was higher in patients with NHL compared with patients in the control group (16.4 ng/mL; P = 0.05). High S-endostatin levels were associated with advanced disease stage (P < 0.0001) and high serum VEGF levels at diagnosis (P = 0.017). The median 5-year survival rate for patients who had S-endostatin concentrations within the highest tertile (> 36.0 ng/mL) was only 34% compared with 57% in patients who had lower S-endostatin levels (P = 0.019). A high S-endostatin level also was associated with a poor outcome in patients with large cell diffuse and immunoblastic lymphoma, which was the largest subgroup within the series (n = 60 patients). Patients who had high pretreatment levels of both S-VEGF and S-endostatin had particularly poor outcomes. High S-endostatin levels had an independent, adverse influence on survival it was entered as a factor into a multivariate analysis together with the factors included in the International Prognostic Index (relative risk, 1.80; 95% confidence interval, 1.08-2.98; P = 0.0024). CONCLUSIONS: High pretreatment levels of S-endostatin are associated with high serum VEGF levels and poor survival in patients with NHL. Prospective studies are warranted to establish the clinical value of longitudinal S-endostatin measurements. PMID- 12767090 TI - A pilot study of transcatheter arterial interferon embolization for patients with hepatocellular carcinoma. AB - BACKGROUND: Systemic, high-dose interferon-alpha treatment given three times per week subcutaneously induces tumor regression in approximately 30% of patients with inoperable hepatocellular carcinoma (HCC). The objective of the current study was to determine the efficacy and safety of transcatheter arterial interferon embolization for the treatment of patients with inoperable HCC. METHODS: Eighteen patients with inoperable HCC were recruited to receive 3 different doses of interferon-alpha-2b (10 megaunits [MU]/m(2), 30 MU/m(2), or 50 MU/m(2)) at intervals of 8-12 weeks. Their tumor response, adverse events, and survival were monitored. RESULTS: In 14 patients with nondiffuse HCC, complete responses and partial responses (> 50% tumor reduction) were observed in 28.6% and 35.7% of patients, respectively. One of four patients with diffuse HCC had a partial response. Thirty-eight percent of patients had normalization of their alpha-fetoprotein level. The median ferritin level at the last follow-up was reduced significantly (765 pmol/L; range, 457-2720 pmol/L) compared with the baseline level (1980 pmol/L; range, 1100-3300 pmol/L; P = 0.011). The median survival was 15.9 months. Transient fever and rigor were the most common side effects observed. Five patients (27.8%) developed hypothyroidism. No significant liver decompensation was observed. CONCLUSIONS: This pilot study showed that transcatheter arterial interferon embolization was an effective method for the treatment of patients with inoperable HCC without significant hepatic toxicity. PMID- 12767091 TI - Neoadjuvant photodynamic therapy as a new approach to treating hilar cholangiocarcinoma: a phase II pilot study. AB - BACKGROUND: Only 20-30% of patients with hilar cholangiocarcinomas (CC) are candidates for potentially curative resection. However, even after curative (R0) resection, these patients have a disease recurrence rate of up to 76%. The current prospective Phase II study investigated photodynamic therapy (PDT) as a neoadjuvant treatment for CC. METHODS: Seven patients with advanced proximal bile duct carcinoma were evaluated. Patients were treated with PDT at the area of tumor infiltration and 2 cm beyond and underwent surgery after a median period of 6 weeks (range, 3-44 weeks). RESULTS: One patient had a Bismuth-Corlette Type II tumor, two patients had Type IIIa, one patient had Type IIIb, and three patients had Type IV. Cholestasis parameters after PDT decreased significantly. No relevant adverse events from PDT occurred except for minor intraoperative phototoxicity in one patient. Three patients underwent right-sided liver resections, two patients underwent left-sided liver resections, and one patient received a combined hilar resection with partial pancreatoduodenectomy (PD) due to tumor extension into the distal bile duct. Liver transplantation and PD were performed in another patient. In all patients, R0 resection was achieved. Four patients developed minor surgical complications, even though the bilioenteric anastomoses were sewn to PDT-pretreated bile ducts. No viable tumor cells were found in the inner 4 mm layer of the surgical specimens. The PDT-pretreated epithelium of the tumor-free proximal resection margins exhibited only minimal inflammatory infiltration. Tumors recurred in 2 patients 6 and 19 months after surgery. The 1-year recurrence free survival rate was 83%. CONCLUSIONS: Neoadjuvant PDT for hilar CC is a low-risk procedure with efficient selective destruction of the superficial 4 mm layer of bile duct tumor without complications exceeding series without neoadjuvant PDT. Neoadjuvant PDT should be evaluated prospectively to determine whether it reduces the rate of local disease recurrence after potentially curative resection. PMID- 12767092 TI - Gemcitabine combined with carboplatin in patients with malignant pleural mesothelioma: a multicentric phase II study. AB - BACKGROUND: Malignant pleural mesothelioma (MPM) is increasing rapidly worldwide. Currently, pemetrexed plus cisplatin chemotherapy showed a survival advantage versus cisplatin alone. No impact on patient survival of surgery, radiotherapy, or their combination has been demonstrated. METHODS: Eight centers in northeastern Italy participated in a Phase II multicenter study. Chemotherapy was comprised of carboplatin area under the concentration-time curve 5 on Day 1 and gemcitabine 1000 mg/m(2) on Days 1, 8, and 15. This cycle was repeated every 4 weeks. RESULTS: Between July 1996 and September 2000, 50 patients were treated. Of the sample, 68% were males, 88% had a Eastern Cooperative Oncology Group performance status score of 0-1, 56% had Stage I-II disease, 68% had epithelioid histology, and 62% had no previous treatments. The delivered dose intensity of gemcitabine was 617 mg/m(2) per week, which was 82% of the planned dose (750 mg/m(2) per week). For carboplatin, the delivered dose intensity was 80 mg/m(2) per week. Overall, 44% of 15th day doses were omitted or reduced. Twenty-six percent of the patients had partial responses (95% confidence interval: 15-40%) and 24% had disease progression. None of the patients had complete responses. The median response duration was 55 weeks (range, 13-113 weeks). Patients had good clinical benefit. For example, 46% had improved dyspnea, 40% improved in weight, and 26% experienced pain reduction. Patients developed Grade 3-4 leukopenia during 18 cycles (11%) of chemotherapy. Grade 3-4 thrombocytopenia occurred more frequently, i.e., there were 24 episodes (15%) among 17 patients. Grade 3 anemia developed among patients during eight cycles (5%). None of the patients developed Grade 3-4 nonhematologic toxicity. The median survival of this sample of patients was 66 weeks with 53%, 30%, and 20% of patients alive at 1, 2, and 3 years, respectively. The median progression-free survival period was 40 weeks. CONCLUSIONS: The gemcitabine/carboplatin combination is a valid option in the treatment of MPM due to its acceptable toxicity profile, the good response rate, and the clinical benefit to patients. Minor adjustments in schedule (3-week cycles instead of 4-week cycles) would permit a more optimal treatment administration. PMID- 12767093 TI - cDNA microarray analysis of gene expression in pathologic Stage IA nonsmall cell lung carcinomas. AB - BACKGROUND: The relationship between altered gene expression and tumor progression in lung carcinoma has yet to be characterized. Gene expression in pathologic Stage IA nonsmall cell lung carcinoma specimens was analyzed using a cDNA microarray. METHODS: Surgical specimens were used for the current study. The pathologic stage was IA (AJCC) in five tumors, IB in two, IIA in one, IIIA in one, and IIIB in one. Seven tumor specimens were adenocarcinomas and three were squamous cell carcinomas. Paired mRNAs from carcinoma cells and normal lung tissue specimens from the same lobe were labeled with different fluorochromes during cDNA probe synthesis in a reverse-transcription reaction. Both synthesized, labeled cDNA probes were mixed and hybridized to the microarray. The signal intensity of each spot was measured by laser scanner and gene expression was quantified as the tumor-to-normal fluorescence ratio (T:N ratio). The gene was overexpressed when the T:N ratio was greater than 2.0 and underexpressed when the ratio was less than 0.5. RESULTS: Overall, 40 (9.4%) of the 425 genes evaluated were overexpressed, and 74 genes (17.4%) were underexpressed. In the 5 Stage IA tumor specimens, 31 (7.3%) genes were overexpressed and 76 (17.9%) were underexpressed. For 30 genes (7.1%), expression was different in Stage IA tumor specimens compared with more advanced tumor specimens. CONCLUSIONS: The cDNA microarray system showed that numerous alterations of gene expression were present in early-stage nonsmall cell lung carcinoma specimens. PMID- 12767095 TI - Cyclooxygenase-2 expression in human pituitary tumors. AB - BACKGROUND: Cyclooxygenase-2 (COX-2) plays a role in progression of colon, breast, pancreas, and lung carcinomas. The authors investigated COX-2 expression in pituitary tumors. METHODS: Expression of COX-2 was evaluated in 164 surgically removed human pituitary tumors. Correlation of COX-2 with MIB-1, a cell proliferation marker, as well as angiogenesis, patient age, gender, tumor type, size, invasiveness, and metastatic potential was investigated. RESULTS: Cyclooxygenase-2 immunoreactivity was confined to the cytoplasm of tumor cells, whereas the nuclei were unlabeled. Few normal peritumoral adenohypophysial cells showed slight COX-2 cytoplasmic immunoreactivity. The staining intensity and the percentage of immunopositive cells were higher in tumors. Most pituitary tumors (96%) were COX-2-immunopositive. Expression was strong in 60 (44%), moderate in 39 (28%), and weak in 32 (24%). Male gonadotroph adenomas and null cell adenomas showed a high level of COX-2 expression. Growth hormone-producing adenomas, prolactin-producing adenomas, thyrotropic hormone-producing adenomas, female gonadotroph adenomas, silent adrenocorticotropic hormone-producing adenomas, and silent subtype 3 adenomas had a low level of COX-2 expression. Significant correlation was demonstrated with patient age, but not with tumor size, invasiveness, and MIB-1 labeling indices. Expression was medium to high in 76% of macroadenomas and in only 45% of microadenomas. Strong correlations were noted with angiogenesis markers, such as microvessel density and surface density. CONCLUSIONS: Correlation with angiogenesis suggests that COX-2 may be involved in the regulation of angiogenesis in pituitary tumors. Phamacologic inhibition of COX-2 activity might suppress angiogenesis in pituitary tumors and may provide a novel approach for medical therapy. PMID- 12767096 TI - Therapeutic effectiveness of novel 5-fluorouracil-loaded poly(methylidene malonate 2.1.2)-based microspheres on F98 glioma-bearing rats. AB - BACKGROUND: Drug delivery to the central nervous system (CNS) remains a real challenge for neurosurgeons and neurologists, because many molecules cannot cross the blood-brain barrier (BBB). In recent years, solid polymeric materials have been implanted into animal and human brains either by surgery or using stereotactic methods to assure the controlled release of a drug over a determined period, thus circumventing the difficulties posed by the BBB. Poly(methylidene malonate 2.1.2) (PMM 2.1.2) is a new polymer that was described a few years ago and that allows the fabrication of novel, 5-fluorouracil (5-FU)-loaded PMM 2.1.2 microspheres. The objective of the current study was to assess the therapeutic effectiveness of those particles in a rat brain tumor model, the F98 glioma. METHODS: Forty-three rats were used in this study. First, a histologic evaluation of the F98 tumor model was performed on Fischer female rats. Thereafter, different groups of rats were injected and were treated with 5-FU microspheres in 2 different suspension media: carboxymethylcellulose (CMC) aqueous solution with or without 5-FU. RESULTS: The tumor was confirmed as extremely aggressive and invasive, even in early development. The 5-FU-loaded microspheres improved rat median survival significantly compared with untreated animals, CMC-treated animals, and 5-FU solution-treated animals when injected in CMC without 5-FU, demonstrating the interest of a sustained release and the efficacy of intratumoral chemotherapy against an established tumor. CONCLUSIONS: PMM 2.1.2 microspheres appeared to be a promising system, because their degradation rate in vivo was longer compared with many polymers, and they may be capable of long-term delivery. PMID- 12767094 TI - Serum S100beta: a noninvasive marker of blood-brain barrier function and brain lesions. AB - BACKGROUND: S100beta protein is expressed constitutively by brain astrocytes. Elevated S100beta levels in cerebrospinal fluid and serum reported after head trauma, subarachnoid hemorrhage, and stroke were correlated with the extent of brain damage. Because elevated serum S100beta also was shown to indicate blood brain barrier (BBB) dysfunction in the absence of apparent brain injury, it remains unclear whether elevation of serum levels of S100beta reflect BBB dysfunction, parenchymal damage, or both. METHODS: The authors conducted a prospective study of serum S100beta levels in six patients who underwent hyperosmotic BBB disruption (BBBD) with intraarterial chemotherapy for primary central nervous system lymphoma. In addition, 53 serum S100beta samples were measured in 51 patients who had a variety of primary or metastatic brain lesions at the time of neuroimaging. RESULTS: S100beta was correlated directly with the degree of clinical and radiologic signs of BBBD in patients who were enrolled in the hyperosmotic study. In patients with neoplastic brain lesions, gadolinium enhancement on a magnetic resonance image was correlated with elevated S100beta levels (n = 45 patients; 0.16 +/- 0.1 microg/L; mean +/- standard error of the mean) versus nonenhancing scans (n = 8 patients; 0.069 +/- 0.04 microg/L). Primary brain tumors (n = 8 patients; 0.12 +/- 0.08) or central nervous system metastases also presented with elevated serum S100beta levels (n = 27 patients; 0.14 +/- 0.34). Tumor volume was correlated with serum S100beta levels only in patients with vestibular schwannoma (n = 6 patients; 0.13 +/- 0.10 microg/L) but not in patients with other brain lesions. CONCLUSIONS: S100beta was correlated directly with the extent and temporal sequence of hyperosmotic BBBD, further suggesting that S100beta is a marker of BBB function. Elevated S100beta levels may indicate the presence of radiologically detectable BBB leakage. Larger prospective studies may better determine the true specificity of S100beta as a marker for BBB function and as an early detection or follow-up marker of brain tumors. PMID- 12767097 TI - Role of adjuvant cryosurgery in intralesional treatment of sacral tumors. AB - BACKGROUND: Cryosurgery is an adjuvant surgical technique for the treatment of benign aggressive, low-grade malignant and metastatic tumors of long bones. It has been used rarely to treat sacral tumors, mainly because of potential damage to nerves, blood vessels, and intrapelvic organs. The authors described their experience with this procedure and provided medium and long-term follow-up results. METHODS: Fifteen procedures of cryosurgery of the sacrum were performed in 14 patients to improve the therapeutic outcome of a variety of tumors. The patient group included 7 males and 7 females with a mean age of 42 +/- 24 years. Three patients were younger than 20 years of age. The procedures were performed at the Tel Aviv Sourasky Medical Center between January 1991 and January 1999. There were seven benign aggressive lesions (four giant cell tumors and three aneurysmal bone cysts), one benign schwannoma, one low-grade chondrosarcoma, five metastatic carcinomas, and one high-grade Ewing sarcoma, all localized at level S(2) or higher. Eight of the bone tumors also involved significant anterior or posterior soft tissue. All patients had severe preoperative pain radiating to the buttocks, perineum, and lower limbs and 9 (64%) patients had bladder and/or rectal dysfunction. Invasive diagnostic procedures and radiation (if warranted) preceded surgery. Sacral posterior fenestration and burr drilling were followed by two-cycle cryosurgery using the open pour technique or the argon-helium-based heat-freeze system. RESULTS: All interventions were performed under combined general and regional anesthesia and concluded uneventfully with moderate blood loss. Thirteen patients were discharged home after 8 +/- 5 days (one patient remained hospitalized for 30 days). Only two patients experienced local disease recurrence during a 3-11-year follow-up period: one was retreated successfully by cryosurgery and the other underwent sacrectomy and radiotherapy elsewhere, with a subsequent loss of visceral functions. No patient suffered chronic pain, deep wound infections, or significant neurologic deficits and all were satisfied with the esthetic outcome. CONCLUSIONS: Cryosurgery is a conservative, feasible, and safe adjuvant technique in the treatment of sacral tumors. It is associated with minimal permanent neurologic and vascular injury compared with sacrectomy. PMID- 12767098 TI - Malignant fibrous histiocytoma and other fibrohistiocytic tumors in pediatric patients: the St. Jude Children's Research Hospital experience. AB - BACKGROUND: Malignant fibrous histiocytoma (MFH) is a controversial entity. In the current study, the authors reviewed their institutional experience with these tumors to characterize their clinical features in pediatric patients and assess the impact of surgical resection on outcome. METHODS: The records of the 28 patients who were diagnosed with MFH or MFH variants of soft tissue between January 1971 and December 2000 were reviewed and the tumors were reclassified according to the World Health Organization guidelines. RESULTS: Seventeen patients had MFH; 10 patients had angiomatoid fibrous histiocytoma (FH), and 1 patient had a plexiform fibrohistiocytic tumor. The median age of patients at the time of diagnosis was 7.3 years. The most common primary tumor site was the extremity (n = 14). Metastatic disease (to the lung) was present in only three patients, each of whom had MFH. Of the 17 MFH tumors, 13 were high grade, 8 were invasive, and 6 measured > 5 cm. All angiomatoid FH tumors and the plexiform fibrohistiocytic tumor were noninvasive, and 10 measured < or = 5 cm. Surgical treatment was comprised of wide local excision with clear margins (n = 18), amputation (n = 3), excision with positive or indeterminate surgical margins (n = 4), partial resection (n = 2), or biopsy only (n = 1). Primary reexcision was performed for 21 patients. The 5-year survival and event-free survival (EFS) estimates for patients with MFH were 76.5% +/- 11.2% and 70.6% +/- 12.1%, respectively; the 5-year survival and EFS estimates were 100% +/- 0% for patients with angiomatoid FH or plexiform fibrohistiocytic tumor. Compared with partial resection or excision, wide local excision or amputation was found to have a positive impact on the probability of EFS in patients with localized disease (P = 0.008). All four patients with metastatic or unresectable MFH had died by the time of last follow-up. CONCLUSIONS: MFH should be distinguished from angiomatoid FH and plexiform fibrohistiocytic tumors, both of which are less aggressive. Wide local excision is the treatment of choice, regardless of the histology or grade of the tumor. Patients with metastatic or unresectable MFH appear to have a poor outcome and would benefit from more effective therapies. PMID- 12767099 TI - Results of a 2-arm Phase II study of 9-nitrocamptothecin in patients with advanced soft-tissue sarcomas. AB - BACKGROUND: The authors conducted a 2-arm Phase II trial of 9-nitrocamptothecin (9-NC), an oral topoisomerase I inhibitor, to define response rates in patients with gastrointestinal (GI) leiomyosarcomas and other soft-tissue sarcomas (STS). METHODS: Patients were required to provide informed consent and have measurable disease, Zubrod performance status < or = 2, and adequate organ function. 9-NC was administered orally at 1.5 mg/m(2) per day x 5 days every week. Response evaluation was performed at 8 weeks, and those with stable or responding disease continued treatment until maximal response was achieved. A total of 56 patients (30 females and 26 males) with a median age of 55 years (range, 19-79 years) were enrolled on the study. Seventeen patients were enrolled on the GI leiomyosarcoma arm; only 1 minor response, lasting < 8 weeks in a patient with liver metastases, was noted, and so this arm was terminated. Thirty-nine patients were entered on the other STS arm. RESULTS: Three patients achieved a partial response (response rate, 8%) for durations of 4, 6, and 13 months, respectively. Fourteen patients had stable disease for a median of 4 months (range, 2-8 months). Two patients died of disease during the first 2 months. Four other patients required hospitalization for nausea, vomiting, and dehydration. Other toxicities included diarrhea (36 patients, 5 with Grade 3 toxicity); fatigue (42 patients, 11 with Grade 3 toxicity); anorexia (32 patients, 1 with Grade 3 toxicity); nausea (37 patients, 2 with Grade 3 toxicity); vomiting (24 patients, 3 with Grade 3 toxicity); neutropenia (14 patients, 5 with Grade 3 toxicity); and thrombocytopenia (16 patients, 5 with Grade 3 or 4 toxicity). CONCLUSIONS: 9-NC is well tolerated but inactive in GI leiomyosarcomas and has minimal activity in previously treated patients with STS. PMID- 12767100 TI - Racial disparities in breast carcinoma survival rates: seperating factors that affect diagnosis from factors that affect treatment. AB - BACKGROUND: Black females have lower breast carcinoma survival rates compared with white females. One possible reason is that black females have more advanced stage breast disease. Another factor may be racial differences in the utilization of cancer treatments. METHODS: The authors determined racial differences in 6 year stage specific survival rates, adjusting for age and treatments (using estrogen receptor [ER] status), to determine whether there were racial differences in treatment. Racial differences in the stage distributions of breast disease were used to examine the impact of racial factors on breast carcinoma diagnosis. RESULTS: For all breast carcinoma cases, the stage specific 6-year survival rates, in general, were significantly lower for black females for all stages combined and for Stages I-III in every age group. However, examination by different treatments, as measured by ER status, revealed some different results. Only black women younger than age 50 years with ER-positive tumors and women younger than age 65 years with ER-negative tumors had significantly lower stage specific survival rates. In addition, the stage distribution analyses showed that black females of every age group had less Stage I breast disease. CONCLUSIONS: For younger black women (younger than age 50 years), there was evidence of racial differences in treatment for both women with ER-positive tumors and women with ER negative tumors, as indicated by their lower stage-specific survival rates. In contrast, for black females age 65 years or older with ER-positive or ER-negative tumors, the lack of a significant difference in the stage-specific survival rate suggests that Medicare may help to alleviate racial disparities in cancer treatment. Furthermore, racial differences in the stage distributions indicated the need for earlier diagnosis for black females of every age. PMID- 12767101 TI - Perceptions and attitudes of clinical oncologists on complementary and alternative medicine: a nationwide survey in Japan. AB - BACKGROUND: The prevalence of complementary and alternative medicine (CAM) is increasing worldwide because of the growing public interest in natural or holistic therapies and because of the flow of information through the Internet. However, there is a lack of communication between cancer patients and their physicians on topics relating to CAM. The authors performed a cross-sectional survey to evaluate the perceptions and attitudes of Japanese clinical oncologists toward cancer CAM. METHODS: The CAM questionnaires were sent to 2118 clinical oncologists. The questionnaires gathered data on background (age, gender, years in practice, specialty, and knowledge of cancer CAM), perception (effectiveness/ineffectiveness, scientific evidence, and drug interactions), and attitude (experience with and response to CAM users). Questions about oncologists' perceptions and attitudes to CAM were limited to herbs and other natural products that were sold over the counter. RESULTS: One hundred sixty-six questionnaires were returned as undeliverable. Of the remaining questionnaires, 751 were returned (a response rate of 39%). Two-thirds of the responders were surgical oncologists and most of the remaining responders were medical oncologists. The majority of oncologists (82%) believed that CAM products were ineffective against cancer. The main reason for this belief was a lack of reliable information (as cited by 85% of oncologists). Only 13% of oncologists had experienced CAM-associated disease improvement in their cancer patients. Of all the oncologists, 84% considered the possibility of drug interactions between anticancer drugs and CAM products. The majority of oncologists (80%) replied that they could neither promote the use of CAM products nor recommend quitting the products, when they were asked about the use of CAM products by cancer patients. CONCLUSIONS: Negative perceptions of CAM products persist among clinical oncologists. A lack of proven effectiveness of CAM products and concerns about drug interactions with anticancer treatment suggest a need for both accurate information on CAM products and clinical trials. PMID- 12767102 TI - Congestive heart failure in patients treated with doxorubicin: a retrospective analysis of three trials. AB - BACKGROUND: Doxorubicin is a highly effective and widely used cytotoxic agent with application that is limited by cardiotoxicity related to the cumulative dose of the drug. A large-scale study that retrospectively evaluated the cardiotoxicity of doxorubicin reported that an estimated 7% of patients developed doxorubicin-related congestive heart failure (CHF) after a cumulative dose of 550 mg/m(2). To assess whether this estimate is reflective of the incidence in the broader clinical oncology setting, the authors evaluated data from three prospective studies to determine both the incidence of doxorubicin-related CHF and the accumulated dose of doxorubicin at which CHF occurs. METHODS: A group of 630 patients who were randomized to a doxorubicin-plus-placebo arm of three Phase III studies, two studies in patients with breast carcinoma and one study in patients with small cell lung carcinoma, were included in the analysis. RESULTS: Thirty-two of 630 patients had a diagnosis of CHF. Analysis indicated that an estimated cumulative 26% of patients would experience doxorubicin-related CHF at a cumulative dose of 550 mg/m(2). Age appeared to be an important risk factor for doxorubicin-related CHF after a cumulative dose of 400 mg/m(2), with older patients (age > 65 years) showing a greater incidence of CHF compared with younger patients (age < or = 65 years). In addition, > 50% of the patients who experienced doxorubicin-related CHF had a reduction < 30% in left ventricular ejection fraction (LVEF) while they were on study. CONCLUSIONS: Doxorubicin related CHF occurs with greater frequency and at a lower cumulative dose than previously reported. These findings further indicate that LVEF is not an accurate predictor of CHF in patients who receive doxorubicin. PMID- 12767103 TI - Nausea and emesis remain significant problems of chemotherapy despite prophylaxis with 5-hydroxytryptamine-3 antiemetics: a University of Rochester James P. Wilmot Cancer Center Community Clinical Oncology Program Study of 360 cancer patients treated in the community. AB - BACKGROUND: Clinical reports suggest that nausea remains a side effect of chemotherapy despite widespread use of serotonin receptor antagonists. This study summarized the frequency, timing, and intensity of postchemotherapy nausea for patients receiving doxorubicin, cisplatin, or carboplatin. METHODS: Three hundred sixty chemotherapy-naive patients (73% female) were enrolled in a study testing the ability of an information intervention to reduce nausea. Of these, 322 subjects completed the Morrow Assessment of Nausea and Emesis (MANE), as well as a 5-day self-report diary, at Cycle 1 (300 subjects completed the MANE and self report diary at Cycle 2). All patients received a 5-hydroxytryptamine-3 receptor antagonist (ondansetron) with dexamethasone on the day of treatment. RESULTS: Seventy-six percent of the patients developed nausea during the 5-day period, beginning with the Cycle 1 infusion, and 73% of patients reported delayed nausea (DN) during Days 2-5. The proportions were similar during Cycle 2. Fifty-five percent of patients described their DN as being of moderate or greater intensity compared with 28% of patients who described acute nausea. Carboplatin was less likely to cause DN than either of the other agents (56% of 106 patients compared with 75% of 47 receiving cisplatin and 83% of 169 taking doxorubicin). The mean peak DN severity was 4.34 (range, 1-7) for doxorubicin, which was significantly higher than the mean peak value for carboplatin (3.66) but was not significantly different from the mean peak value for cisplatin (4.26). Eighteen percent of patients did not experience nausea until Day 3 or later. CONCLUSIONS: Despite prophylaxis with ondansetron, the majority of patients receiving one of these common chemotherapy agents experienced nausea. The frequency of DN was nearly twice that of acute nausea. Results show the need for continued development of antiemetics that are effective against DN. PMID- 12767104 TI - Detection of micrometastasis of neuroblastoma to bone marrow and tumor dissemination to hematopoietic autografts using flow cytometry and reverse transcriptase-polymerase chain reaction. AB - BACKGROUND: The identification of neuroblastoma metastases to bone marrow (BM) is requisite in staging disease for risk-adopted therapy. However, micrometastases were not elucidated fully. METHODS: Flow cytometry (FCM) with CD45/CD56/CD81 and reverse transcriptase-polymerase chain reactions (RT-PCR) for tyrosine hydroxylase (TH) transcripts were used to evaluate neuroblastoma in bilateral BM aspirates at diagnosis, BM autografts, peripheral blood stem cell (PBSC) collections, and CD34(+) cell products of 27 children. RESULTS: TH transcripts were amplified in histology-negative (H(-)) BM specimens from seven patients (four patients with Stage 3 disease, two with Stage 4 disease, and one with Stage 4S disease), revealing a prevalence of submicroscopic metastasis. The median number of CD45(-)CD81(+)CD56(+) cells in four H(-) TH(-) BM samples from two patients with Stage 1 and Stage 2 disease, respectively, was comparable to that encountered in 10 normal BM controls (0.003% [range, 0.002-0.004%] vs. 0.004% [0 0.008%], P = 0.724). In six H(-) TH(+) BM specimens from three patients whom were otherwise diagnosed with neuroblastoma Stage 3, 0.031% (0.009-0.06%) CD45( )CD81(+)CD56(+) cells were detected. Besides, 1.474% (0.088-3.009%) CD45( )CD81(+)CD56(+) cells were identified in four H(-) TH(+) BM specimens from two patients at Stage 4. TH transcripts were evident in four of five BM autografts and in 22 of 45 (48.9%) PBSC specimens. FCM demonstrated 0.018% and 0.049% CD45( )CD81(+)CD56(+) cells in two TH(+) BM autografts, respectively. The number of CD45(-)CD81(+)CD56(+) cells was higher in 19 TH(+) PBSC specimens than in 20 TH( ) PBSC specimens (0.026% [0.006-1.128%] vs. 0% [0-0.009%], P < 0.0001). CD34(+) cell selection achieved 2.9 (2.1-3.5) log depletion of CD45(-)CD81(+)CD56(+) cells in four manipulated products, rendering six of seven PBSC autografts TH free. CONCLUSIONS: FCM in combination with RT-PCR evaluated neuroblastoma micrometastasis and assessed the purity of hematopoietic autografts for transplant. However, the clinical relevance remains to be elucidated. PMID- 12767105 TI - Dexamethasone-associated toxicity during induction chemotherapy for childhood acute lymphoblastic leukemia is augmented by concurrent use of daunomycin. AB - BACKGROUND: The goals of the current study were to examine the incidence and severity of toxicity resulting from dexamethasone and prednisone during induction therapy for children with precursor B-cell acute lymphoblastic leukemia (ALL) and to determine whether the addition of daunomycin affected toxicity. METHODS: Medical records of patients with precursor B-cell ALL from January 1996 through June 2000 were reviewed retrospectively for toxicity during the 4-week induction phase and the 2 weeks after the induction phase. RESULTS: One hundred seventy-six patients age < 14 years were diagnosed with precursor B-cell ALL from January 1996 through June 2000. Of the 156 evaluable patients, 106 were treated with prednisone and 50 with dexamethasone. Fifty-two patients received steroids, L asparaginase, and vincristine, whereas 104 high-risk patients received daunomycin in addition to these 3 agents. The incidence of gastritis was significantly higher among patients receiving dexamethasone (P = 0.01); incidence rates of hyperglycemia, hypertension, and myopathy were similar for all treatment groups. Dexamethasone led to more weight gain than did prednisone (+11.9% vs. +5.4%; P = 0.002). Serious infections were observed in 27 (25.5%) and 18 (36%) patients receiving prednisone and dexamethasone, respectively (P < or = 0.2). Five patients, four of whom received prednisone and one of whom received dexamethasone, died of infection. The addition of daunomycin to treatment regimens increased overall toxicity (P < 0.01). When daunomycin was included in treatment regimens, toxicity was greater among patients receiving dexamethasone; in contrast, when daunomycin was not included, toxicity was equal for both treatment groups. Regardless of daunomycin use, there was no difference in the incidence of serious infection between the two groups. ALL treatment was not compromised by steroid-related toxicity in either group. CONCLUSIONS: The addition of daunomycin led to a much larger increase in dexamethasone-related toxicity compared with the increase in prednisone-related toxicity. Although the use of daunomycin enhanced dexamethasone-related toxicity, this enhancement did not result in a higher mortality rate or the alteration of planned ALL therapy. PMID- 12767106 TI - Disclosure of the right of research participants to receive research results: an analysis of consent forms in the Children's Oncology Group. AB - BACKGROUND: The offer of return of research results to study participants has many potential benefits. The current study examined the offer of return of research results by analyzing consent forms from 2 acute lymphoblastic leukemia studies of the 235 institutional members of the Children's Oncology Group. METHODS: Institutional review board (IRB)-approved consent forms from 2 standard risk acute lymphoblastic leukemia studies (Children's Cancer Group [CCG] 1991 and Pediatric Oncology Group [POG] 9407) were analyzed independently by 2 reviewers. RESULTS: The authors received replies from 202 of the 235 institutions that were contacted (85%). One hundred eighty-one institutions had CCG 1991 (n = 96) or POG 9905 (n = 85) protocols that were approved by an IRB. Most institutions provided contact information for the principal investigator (n = 175; 97%) and a member of the institution's research services office (n = 154; 85%). Only 5 (2.8%) institutions provided an indication of a participant's right to receive a summary of research results; most of these institutions provided details on how (n = 5) or when (n = 5) this was to occur. All of these institutions (n = 162; 89.5%) provided a specific statement offering new information that might affect a participant's decision to continue to participate in a study. Only 2 institutional consent forms offered participants the option to receive research results, and only 10 (5.5%) consent forms contained an unambiguous, specific statement offering to provide new information after the study was closed. CONCLUSIONS: Few institutional review board-approved consent forms explicitly indicate the right of research recipients to receive a summary of the results of the research in which they have participated. PMID- 12767107 TI - The memorial anxiety scale for prostate cancer: validation of a new scale to measure anxiety in men with with prostate cancer. AB - BACKGROUND: The psychological difficulties facing men with prostate cancer are acknowledged widely, yet identifying men who may benefit from mental health treatment has proven to be a challenging task. The authors developed the Memorial Anxiety Scale for Prostate Cancer (MAX-PC) to facilitate the identification and assessment of men with prostate cancer-related anxiety. This scale consists of three subscales that measure general prostate cancer anxiety, anxiety related to prostate specific antigen (PSA) levels in particular, and fear of recurrence. METHODS: Ambulatory men with prostate cancer (n = 385 patients) were recruited from clinics throughout the United States. Prior to routine PSA tests, participants completed a baseline assessment packet that included the Hospital Anxiety and Depression Scale; the Distress Thermometer; the Functional Assessment of Cancer Therapy Scale, Prostate Module; and measures of role functioning, sleep, and urinary functioning. PSA values from the last three tests also were collected. Follow-up evaluation was completed within 2 weeks after patients learned of their PSA test result using a subset of these scales. RESULTS: Analysis of the MAX-PC revealed a high degree of internal consistency and test retest reliability for the total score and for the three subscales, although reliability was somewhat weaker for the PSA Anxiety Scale. Concurrent validity was demonstrated by correlations between the MAX-PC and measures of anxiety. Overall changes in PSA levels were correlated only modestly with changes in MAX PC scores (correlation coefficient, 0.13; P = 0.02). CONCLUSIONS: The MAX-PC appears to be a valid and reliable measure of anxiety in men with prostate cancer receiving ambulatory care. PMID- 12767108 TI - Are the symptoms of cancer and cancer treatment due to a shared biologic mechanism? A cytokine-immunologic model of cancer symptoms. AB - BACKGROUND: Cancers and cancer treatments produce multiple symptoms that collectively cause a symptom burden for patients. These symptoms include pain, wasting, fatigue, cognitive impairment, anxiety, and depression, many of which co occur. There is growing recognition that at least some of these symptoms may share common biologic mechanisms. METHODS: In November 2001, basic and clinical scientists met to consider evidence for a cytokine-immunologic model of symptom expression along with directions for future research. RESULTS: The characteristics of cytokine-induced sickness behavior in animal models have much in common with those of symptomatic cancer patients. Sickness behavior refers to a set of physiologic and behavioral responses observed in animals after the administration of infectious or inflammatory agents or certain proinflammatory cytokines. In some cases, these responses can be prevented by cytokine antagonists. A combination of animal and human research suggests that several cancer-related symptoms may involve the actions of proinflammatory cytokines. CONCLUSIONS: Based on the similarities between cancer symptoms and sickness behavior, the authors discussed approaches to further test the implications of the relationship between inflammatory cytokines and symptoms for both symptom treatment and symptom prevention. PMID- 12767109 TI - Absence of c-Kit and members of the epidermal growth factor receptor family in refractory germ cell cancer. PMID- 12767112 TI - Dmt and opioid peptides: a potent alliance. AB - The introduction of the Dmt (2',6'-dimethyl-L-tyrosine)-Tic pharmacophore into the design of opioid ligands produced an extraordinary family of potent delta opioid receptor antagonists and heralded a new phase in opioid research. First reviewed extensively in 1998, the incorporation of Dmt into a diverse group of opioid molecules stimulated the opioid field leading to the development of unique analogues with remarkable properties. This overview will document the crucial role played by this residue in the proliferation of opioid peptides with high receptor affinity (K(i) equal to or less than 1 nM) and potent bioactivity. The discussion will include the metamorphosis between delta-opioid receptor antagonists to delta-agonists based solely on subtle structural changes at the C terminal region of the Dmt-Tic pharmacophore as well as their behavior in vivo. Dmt may be considered promiscuous due to the acquisition of potent mu-agonism by dermorphin and endomorphin derivatives as well as by a unique class of opioidmimetics containing two Dmt residues separated by alkyl or pyrazinone linkers. Structural studies on the Dmt-Tic compounds were enhanced tremendously by x-ray diffraction data for three potent and biologically diverse Dmt-Tic opioidmimetics that led to the development of pharmacophores for both delta opioid receptor agonists and antagonists. Molecular modeling studies of other unique Dmt opioid analogues illuminated structural differences between delta- and mu-receptor ligand interactions. The future of these compounds as therapeutic applications for various medical syndromes including the control of cancer associated pain is only a matter of time and perseverance. PMID- 12767113 TI - Successful identification of novel agents to control infectious diseases from screening mixture-based peptide combinatorial libraries in complex cell-based bioassays. AB - Mixture-based peptide synthetic combinatorial libraries (SCLs) represent a valuable source for the development of novel agents to control infectious diseases. Indeed, a number of studies have now proven the ability of identifying active peptides from libraries composed of thousands to millions of peptides in cell-based biosystems of varying complexity. Furthermore, progressing knowledge on the importance of endogenous peptides in various immune responses lead to a regain in importance for peptides as potential therapeutic agents. This article is aimed at providing recent studies in our laboratory for the development of antimicrobial or antiviral peptides derived from mixture-based SCLs using cell based assays, as well as a short review of the importance of such peptides in the control of infectious diseases. Furthermore, the use of positional scanning (PS) SCL-based biometrical analyses for the identification of native optimal epitopes specific to HIV-1 proteins is also presented. PMID- 12767115 TI - Grb2 SH2 domain-binding peptide analogs as potential anticancer agents. AB - The growth factor receptor-bound protein 2 (Grb2) plays an important role in the Ras signaling pathway. Several proteins were found to be overexpressed by oncogenes in the Ras signaling pathway, rendering Grb2 a potential target for the design of antitumor agents. Blocking the interaction between the phosphotyrosine containing activated receptor and the Src-homology 2 (SH2) domain of Grb2 thus constitutes an important strategy for the development of potential anticancer agents. X-ray, NMR structural investigations, and molecular modeling studies have provided the target structure of Grb2 SH2 domain-alone or complexed with a phosphotyrosine-containing peptide-which is useful for the structure-based design of peptides or peptidomimetics with high affinity for the Grb2 SH2 domain. We review here the variety of approaches to Grb2 SH2 pepide inhibitors developed with the aim of interrupting Grb2 recognition. Inhibitory effects of peptide analogs on the Grb2 SH2 domain and their binding affinities for Grb2 SH2 were determined by ELISA, cell-based assays, or Surface Plasman Resonance (SPR) technology. Results of theses studies provide important information for further modifications of lead peptides, and should lead to the discovery of potent peptides as anticancer agents. PMID- 12767114 TI - Oligopeptide inhibition of class I ribonucleotide reductases. AB - Class I ribonucleotide reductases (RRs), which are well-recognized targets for cancer chemotherapeutic and antiviral agents, are composed of two different subunits, R1 and R2, and are inhibited by oligopeptides corresponding to the C terminus of R2, which compete with R2 for binding to R1. These peptides specifically inhibit the RRs from which they are derived, and closely homologous RRs, but do not inhibit less homologous RRs. Here we review results obtained for oligopeptide inhibition of RRs from several sources, including related x-ray, NMR, and modeling results. The most extensive studies have been performed on herpes simplex virus-RR (HSV-RR) and mammalian-RR (mRR). A common model fits the data obtained for both enzymes, in which the C-terminal residue of the oligopeptide (Leu for HSV-RR, Phe for mRR) binds with high specificity to a narrow and deep hydrophobic subsite, and two or more hydrophobic groups at the N terminal portion of the peptide bind to a broad and shallow second hydrophobic subsite. The studies have led to the development of highly potent and specific inhibitors of HSV-RR and promising inhibitors of mRR, and indicate possible directions for the development of inhibitors of bacterial and fungal RRs. PMID- 12767116 TI - Synthetic peptide vaccine and antibody therapeutic development: prevention and treatment of Pseudomonas aeruginosa. AB - Pseudomonas aeruginosa and Pseudomonas maltophilia account for 80% of opportunistic infections by pseudomonads. Pseudomonas aeruginosa is an opportunistic pathogen that causes urinary tract infections, respiratory system infections, dermatitis, soft tissue infections, bacteremia, and a variety of systemic infections, particularly in patients with severe burns, and in cancer and AIDS patients who are immunosuppressed. Pseudomonas aeruginosa is notable for its resistance to antibiotics, and is therefore a particularly dangerous pathogen. Only a few antibiotics are effective against Pseudomonas, including fluoroquinolones, gentamicin, and imipenem, and even these antibiotics are not effective against all strains. The difficulty treating Pseudomonas infections with antibiotics is most dramatically illustrated in cystic fibrosis patients, virtually all of whom eventually become infected with a strain that is so resistant that it cannot be treated. Since antibiotic therapy has proved so ineffective as a treatment, we embarked on a research program to investigate the development of a synthetic peptide consensus sequence vaccine for this pathogen. In this review article we will describe our work over the last 15 years to develop a synthetic peptide consensus sequence anti-adhesin vaccine and a related therapeutic monoclonal antibody (cross-reactive to multiple strains) to be used in the prevention and treatment of P. aeruginosa infections. Further, we describe the identification and isolation of a small peptide structural element found in P. aeruginosa strain K (PAK) bacterial pili, which has been proven to function as a host epithelial cell-surface receptor binding domain. Heterologous peptides are found in the pili of all strains of P. aeruginosa that have been sequenced to date. Several of these peptide sequences have been used in the development of an consensus sequence anti-adhesin vaccine targeted at the prevention of host cell attachment and further for the generation of a monoclonal antibody capable of prevention and treatment of existing infections. PMID- 12767117 TI - Homology model of the CB1 cannabinoid receptor: sites critical for nonclassical cannabinoid agonist interaction. AB - Association of cannabimimetic compounds such as cannabinoids, aminoalkylindoles (AAIs), and arachidonylethanolamide (anandamide) with the brain cannabinoid (CB(1)) receptor activates G-proteins and relays signals to regulate neuronal functions. A CB(1) receptor homology model was constructed using the published x ray crystal structure of bovine rhodopsin (Palczewski et al., Science, 2000, Vol. 289, pp. 739-745) in the conformation most likely to represent the "high affinity" state for agonist binding to G-protein coupled receptors (GPCRs). A molecular docking approach that combined Monte Carlo and molecular dynamics simulations was used to identify the putative binding conformations of nonclassical cannabinoid agonists, including AC-bicyclic CP47497 and CP55940, and ACD-tricyclic CP55244. Placement of these ligands was based upon the assumption of a critical hydrogen bond between the A-ring OH and the side chain N of Lys192 in transmembrane helix 3. We evaluated two alternative binding conformations, C3 in and C3-out, denoting the directionality of the ligand C3 side chain within the receptor with respect to the inside or the outside of the cell. Assuming both the C3-in or C3-out conformation, the calculated ligand-receptor binding energy (DeltaE(bind)) was correlated with the experimentally observed binding affinity (K(i)) for a series of nonclassical cannabinoid agonists. The C3-in conformation was marginally better than the alternative C3-out conformation in predicting the rank order of the tested nonclassical cannabinoid analogs. Adopting the C3-in conformation due to the greater number of receptor interactions with known pharmacophoric elements of the ligand, key residues were identified comprising the presumed hydrophobic pocket that interacts with the C3 side chain of cannabinoid agonists. Key hydrogen bonds would form between both K3.28(192) and E(258) and the A-ring OH, and between Q(261) and the C-ring C-12 hydroxypropyl. In summary, the present study represents one of the first attempts to construct a homology model of the CB(1) cannabinoid receptor based upon the published bovine rhodopsin x-ray crystal structure and to elucidate the putative ligand binding site for nonclassical cannabinoid agonists. We postulated sites of the CB(1) receptor critical for the ligand interaction, including the hydrophobic pocket interacting with the key pharmacophoric moiety, the C3 side chain. More work is needed to delineate between two alternative (and possibly other) binding conformations of the nonclassical cannabinoid ligands within the CB(1) receptor. The present study provides a consistent framework for further investigation of the CB(1) receptor-ligand interaction and for the study of CB(1) receptor activation. PMID- 12767118 TI - An autocatalytic reaction as a model for the kinetics of the aggregation of beta amyloid. AB - Alzheimer's disease is the commonest form of senile dementia, affecting almost 20 million people worldwide. This neurodegenerative disorder is characterized by amyloid deposition in senile plaques, composed primarily of fibrils of an aggregated peptide, beta-amyloid. Fibrillation of beta-amyloid is a nucleation dependent polymerization process, which is controlled by two kinetics parameters: the nucleation rate and the elongation or growth rate. As the kinetics of fibrillation is strongly dependent on the presence of trace amounts of fibrils, we suggest that the aggregation of beta-amyloid is a model of autocatalytic reaction. A mathematical analysis, permitting quantitative monitoring of the kinetics of fibrillogenesis of beta-amyloid, nucleation, and elongation constants, is presented. The model was checked by applying it to the aggregation of the fragment 1-40 of the beta-amyloid. Understanding of these rate constants may facilitate the study of the effect of substances used for controlling fibril creation and growth. The disaggregating effect of dodecyl trimethylammonium bromide, a cationic surfactant, was easily quantified by means of the model. PMID- 12767119 TI - Completely different amyloidogenic potential of nearly identical peptide fragments. PMID- 12767120 TI - An improved model for analyzing the small angle x-ray scattering of starch granules. AB - The structure of starch was studied using small-angle x-ray scattering (SAXS). The scattering data was modeled by considering a finite stack of alternating lamellae that are allowed to fluctuate both along the layer repeat direction and along the transverse layer direction. Analysis in this way of the SAXS data from starch allowed fresh insights into the native structure of several starch species, particularly potato starch. The novel model presented in this work was able to capture the experimentally observed SAXS patterns much better than previous models, which did not incorporate transverse layer fluctuations. PMID- 12767121 TI - Acid-induced denaturation of Escherichia coli ribonuclease HI analyzed by CD and NMR spectroscopies. AB - Acid-induced denaturation of the ribonuclease HI protein from Escherichia coli was analyzed by CD and NMR spectroscopies. The CD measurement revealed that the acid denaturation at 10 degrees C proceeds from the native state (N-state) to a molten globule-like state (A-state), through an apparently more unfolded state (U(A)-state). In (1)H-(15)N heteronuclear single-quantum coherence (HSQC) spectra, cross peaks from the N-state and those from the other two states are distinctively observed, while the U(A)-state and A-state are not distinguished from each other. Cross peaks from the U(A)/A-states showed a small pH dependence, which suggests a similarity in the backbone structure between the two states. The direct hydrogen-deuterium (H-D) exchange measurement at pH with the largest population of U(A)-state revealed that at least alpha-helix I is highly protected in the structure of the U(A)-state. A pH-jump H-D exchange analysis showed that the protection of alpha-helix I is highest also in the A-state. The profile of hydrogen-bond protection indicated that the structure of the A-state is closely related to that of the kinetic folding intermediate. PMID- 12767122 TI - Local and long-range structural effects caused by the removal of the N-terminal polypeptide fragment from immunoglobulin L chain lambda. AB - The role of the N-terminal polypeptide fragment of the immunoglobulin l-chain in V domain packing stability, and the flexibility of the whole chain was approached by molecular dynamics simulation. The observations were supported by experimental analysis. The N-terminal polypeptide fragment appeared to be the low-stability packing element in the V domain. At moderately elevated temperature it may be replaced at its packing locus by Congo red and then removed by proteolysis. After removal of Congo red by adsorption to (diethylamino)ethyl (DEAE) cellulose, the stability of complete L chain and of L chain devoid of the N-terminal polypeptide fragment were compared. The results indicated that the N-terminal polypeptide fragment plays an essential role in the stability of the V domain. Its removal makes the domain accessible for ANS and Congo red dye binding without heating. The decreased domain stability was registered in particular as increased root mean square (RMS) fluctuation and higher susceptibility to proteolytic attack. The long-range effect was most clearly manifested at 340 K as independent V and C domain fluctuation in the l-chain devoid of the N-terminal polypeptide fragment. This is likely due to the lack of direct connections between the N- and C-termini of the V domain polypeptide. In a complete V domain the connection involves residues 8-12 and 106-110 in particular. Partial or complete disruption of this connection increases the freedom of V domain rotation, while its increased cohesion strengthens the coupling of the V and C domains, making the whole L chain less flexible. PMID- 12767123 TI - The NMR-derived conformation of neuropeptide AF, an orphan G-protein coupled receptor peptide. AB - The tertiary structure of the pain modulating and anti-opiate neuropeptide, human neuropeptide AF (NPAF) (the sequence is AGEGLNSQFWSLAAPQRF-NH(2)), was determined by (1)H-NMR. The structure of NPAF was determined in two solvent systems, namely 50%/50% trifluoroethanol-d(3)/H(2)O (TFE/H(2)O) and in the cell membrane mimetic micelle, sodium dodecylsulfate-d(25) (SDS). The receptor for NPAF is an orphan G protein coupled receptor, and the micellar SDS solvent system was used to emulate the cell membrane surface in line with the Cell Membrane Compartments Theory proposed by R. Schwyzer (Biopolymers, 1995, Vol. 37, pp. 5-16). In both solvent systems, NPAF was found to be primarily alpha-helical within the central portion of the molecule, from Asn(6) to Ala(14). The N-terminus was random in both solvent systems. In the SDS solution, the C-terminal tetrapeptide was structured and formed a type I beta-turn, whereas in TFE/H(2)O it was unstructured, showing the importance of the C-terminal tetrapeptide in receptor recognition. NPAF was found to associate with SDS, and was shown to be near the surface of the micelle by spin label studies with 5-doxyl-stearic acid. PMID- 12767124 TI - Sequence-dependent solution structure and motions of 13 TATA/TBP (TATA-box binding protein) complexes. AB - The TATA element is a well-known example of a DNA promoter sequence recognized by the TATA box binding protein (TBP) through its intrinsic motion and deformability. Although TBP recognizes the TATA element octamer unusually (through the minor groove, which lacks the distinctive features of the major groove), single base-pair replacements alter transcriptional activity. Recent crystallographic experiments have suggested that TATA/TBP complexes differing by a single base pair retain substantial structural similarity despite their functional differences in activating transcription. To investigate the subtle role of sequence-dependent motion within the TATA element and certain aspects of its effect on assembly of the transcriptional complex, we examine 5-ns dynamics trajectories of 13 variant TATA/TBP complexes differing from each other by a single base pair. They include the wild-type (WT) adenovirus 2 major late promoter (AdMLP) TATA element, TATAAAAG (the octamer specifies positions -31 to 24 with respect to the transcription initiation site), and the variants A31 (i.e., AATAAAAG), T30, A29, C29, G28, T28, T27, G26, T26, C25, T25, and T24. Our simulated TATA/TBP complexes develop sequence-dependent structure and motion trends that may lead to favorable orientations for high-activity variants (with respect to binding TFIIA, TFIIB, and other transcription factors), while conversely, accelerate dissociation of low-activity TATA/TBP complexes. The motions that promote favorable geometries for preinitiation complexes include small rotations between TBP's N- and C-terminal domains, sense strand DNA backbone "slithering," and rotations in TBP's H2 and H2' helices. Low-activity variants tend to translate the H1 and H1' helices and withdraw the intercalating phenylalanines. These cumulative DNA and protein motions lead to a spatial spread of complex orientations up to 4 A; this is associated with an overall bend of the variant TATA/TBP complexes that spans 93 degrees to 110 degrees (107 degrees for the crystal reference). Taken together, our analyses imply larger differences when these local structural and bending changes are extended to longer DNA (upstream and downstream) and suggest that specific local TATA/TBP motions (e.g., shifts in TBP helices and TATA bases and backbone) play a role in modulating the formation and maintenance of the transcription initiation complex. PMID- 12767125 TI - Zinc binding in peptide models of angiotensin-I converting enzyme active sites studied through 1H-NMR and chemical shift perturbation mapping. AB - We report the design and synthesis through solid phase 9-flourenylmethoxycarbonyl (Fmoc) chemistry of the two angiotensin-I converting enzyme active sites possessing the general sequence HEMGHX(23)EAIGDX(3). Their zinc-binding properties were monitored in solution through high-resolution (1)H-NMR. The obtained data were analyzed in terms of chemical shift differences. The results indicate that zinc binds to the HEMGH and the EAIGD characteristic motifs, and suggest possible coordination modes of zinc in the native enzyme. PMID- 12767126 TI - Structural characterization and artificial fiber formation of Bombyx mori silk fibroin in hexafluoro-iso-propanol solvent system. AB - High-resolution solution (13)C-NMR and CD studies of Bombyx mori silk fibroin revealed the presence of an ordered secondary structure 3(10)-helix, in hexafluoro-iso-propanol (HFIP). The solid-state structure of the silk fibroin film prepared by drying it gently from the HFIP solution still keep the structure, 3(10)-helix, which was studied with high-resolution solid state (13)C NMR. The structural transition from the 3(10)-helix to silk II structure, heterogeneous structure including antiparallel beta-sheet, occurred during the artificial spinning from the HFIP solution. The wide-angle x-ray diffraction and differential scanning calorimetry thermograms of the artificial spinning fiber after postspinning treatments were observed together with the stress-strain curves. The results emphasize that the molecular structures, controlled morphology, and mechanical properties of the protein-based synthetic polymers can be modulated for enhancing biocompatibility. PMID- 12767127 TI - Molecular dynamics simulation studies of cytochrome b5 from outer mitochondrial and microsomal membrane. AB - Two forms of cytochrome b(5) have been identified, associated with the outer membrane of liver mitochondria (OM cyt b(5)) and with the membrane of the endoplasmic reticulum (microsomal, Mc cyt b(5)). These proteins have very similar structures, but differ significantly in physical properties, with the OM cyt b(5) exhibiting a more negative reduction potential, higher stability, and stronger interactions with the heme. We perform molecular dynamics simulations to probe the structures and fluctuations of the two proteins in solution, to help explain the observed physical differences. We find that the structures of the two proteins, highly similar in the crystal, differ in position of a surface loop involving residues 49-51 in solution. Hydrophobic residues Ala-18, Ile-32, Leu 36, and Leu-47 tend to cluster together on the surface of rat OM cyt b(5), blocking water access to the protein interior. In bovine Mc cyt b(5), two of these positions, Ser-18 and Arg-47, are occupied by hydrophilic residues. This leads to breaking the hydrophobic cluster and allowing the protein to occupy a more open conformation. A measure of this structural transition is the opening of a cleft on the protein surface, which is 5 A wider in the OM cyt b(5) simulation compared to the Mc form. The OM protein also appears to have a more compact hydrophobic core in its beta-sheet region. These effects may be used to explain observed stability differences between the two proteins. PMID- 12767128 TI - NMR identification of left-handed polyproline type II helices. AB - NMR characteristics of a model left-handed 3(1)-helical peptide are reported in this study. With temperature and sequence corrections on the predicted random coil (15)N chemical shifts, a significant (15)N chemical shift deviation is observed for the model 3(1) peptide. The (15)N chemical shift differences also correlate well with the molar ellipticities (at 220 nm) of the CD spectra at different temperatures, indicating that the (15)N chemical shift is a sensitive probe for 3(1)-helices. The average (3)J(HNalpha) and (1)J(CalphaHalpha) values of the model peptide are determined to be 6.5 and 142.6 Hz, respectively, which are consistent with the values calculated from the geometry of 3(1)-helices. With careful measurements of amide (15)N chemical shifts and incorporating temperature and sequence effect corrections, the (15)N chemical shifts can be used together with (3)J(HNalpha) and (1)J(CalphaHalpha) to differentiate 3(1)-helices from random coils with high confidence. Based on the observed NMR characteristics, a strategy is developed for probing left-handed 3(1)-helical structures from other secondary structures. PMID- 12767129 TI - The signature molecular descriptor. 1. Using extended valence sequences in QSAR and QSPR studies. AB - We present a new descriptor named signature based on extended valence sequence. The signature of an atom is a canonical representation of the atom's environment up to a predefined height h. The signature of a molecule is a vector of occurrence numbers of atomic signatures. Two QSAR and QSPR models based on signature are compared with models obtained using popular molecular 2D descriptors taken from a commercially available software (Molconn-Z). One set contains the inhibition concentration at 50% for 121 HIV-1 protease inhibitors, while the second set contains 12865 octanol/water partitioning coefficients (Log P). For both data sets, the models created by signature performed comparable to those from the commercially available descriptors in both correlating the data and in predicting test set values not used in the parametrization. While probing signature's QSAR and QSPR performances, we demonstrates that for any given molecule of diameter D, there is a molecular signature of height h 8.68 A). The dc magnetic susceptibility of 1 indicates a ferromagnetically coupled S = 4 ground state best fit to the parameters g = 2.23, J = +4.3 cm(-1), and D(Ni) = +8.8 cm(-1) for the Hamiltonian H = -2J [(S(Fe(1)) + S(Fe(2))).(S(Ni(1)) + S(Ni(2)) + S(Ni(3)))] + D[S(Ni(1))(z)(2) + S(Ni(2))(z)(2) + S(Ni(3))(z)(2)]. The extended square molecule [Ni(bpy)(2)(H(2)O)][[Ni(bpy)(2)](2)[Fe(CN)(6)](2)].12H(2)O, (2).12H(2)O (bpy = 2,2'-bipyridine) crystallizes in the space group P1 (No. 2) with unit cell parameters a = 13.264(3) A, b = 17.607(4) A, c = 18.057(4) A, alpha = 94.58(3) degrees, beta = 103.29(3) degrees, gamma = 95.18(3) degrees, V = 4065(2) A(3), and Z = 2. The pi-pi interactions of 3.29 A between the bpy ligands are the closest intermolecular contacts, and the intermolecular M...M separations are greater than 7.76 A. The dc magnetic susceptibility data for 2 are also in accord with an S = 4 ground state arising from intramolecular ferromagnetic coupling. The data were best fit to the parameters g = 2.25, J = J' = +3.3 cm(-1), and D(Ni) = +5.8 cm(-1) for the Hamiltonian H = -2J[(S(Fe(1)) + S(Fe(2))).(S(Ni(1)) + S(Ni(2)))] - 2J'[(S(Fe(2)).S(Ni(3)))] + D[S(Ni(1))(z)(2) + S(Ni(2))(z)(2) + S(Ni(3))(z)(2)]. No evidence for long-range magnetic ordering was observed for crystalline samples of 1 or 2. PMID- 12767177 TI - Examination of novel zinc-binding groups for use in matrix metalloproteinase inhibitors. AB - The tetrahedral zinc complex [(Tp(Ph,Me))ZnOH] (Tp(Ph,Me) = hydrotris(3,5 phenylmethylpyrazolyl)borate) was combined with 1-hydroxy-2(1H)-pyridinone, 3 hydroxy-2(1H)-pyridinone, 3-hydroxy-1-methyl-2(1H)-pyridinone, 3-hydroxy-1,2 dimethyl-4(1H)-pyridinone, 1-hydroxy-2(1H)-pyridinethione, and 3-hydroxy-2-methyl 4-pyrone to generate the complexes [(Tp(Ph,Me))Zn(ZBG)] (ZBG = zinc-binding group). These complexes were synthesized to explore the coordination geometry of potential novel zinc-binding groups for use in matrix metalloproteinase (MMP) inhibitors. The solid-state structures of all six metal complexes were determined by X-ray crystallography. These structures combined with IR and (1)H NMR data demonstrate that these ZBGs bind in a strong, bidentate fashion to the zinc(II) ion. Modeling studies indicate that these ZBGs can easily fit into the MMP active site. In an effort to develop more effective inhibitors of MMPs, this work has revealed molecular-level interactions for six potential new ZBGs. PMID- 12767178 TI - Gallium and indium hydrazides. Molecular and electronic structure of In[N(SiMe3)NMe2]3 and related compounds. AB - Gallium and indium hydrazides, Ga[N(SiMe(3))NMe(2)](3) (1) and In[N(SiMe(3))NMe(2)](3) (2), were synthesized from the reactions of metal chlorides and Li[N(SiMe(3))NMe(2)]. Single crystal X-ray crystallographic analysis revealed that compound 2 was monomeric with trigonal planar geometries on the indium and the indium-bonded nitrogen atoms. The average In[bond]N distance of 2.078(3) A and the N[bond]In[bond]N[bond]N dihedral angles did not provide clear structural evidence of In[bond]N pi-bonding. The electronic absorption spectra of the indium hydrazido complex revealed transitions at significantly lower energies compared to those observed in the tris(amido) compounds, In[N(SiMe(3))(2)](3) (3) and In[N((t)Bu)(SiMe(3))](3) (4). The absorptions of the indium and gallium compounds were attributed to ligand-metal charge transfer transitions. Trends in the electronic transitions for compounds 2 and 3 calculated at the time-dependent density functional and configuration interaction including single excitations levels, both using a minimal basis set, were consistent with the experimental data, and Mulliken charge analyses support the assignment to ligand-to-metal charge transfer transitions. These calculations also demonstrated the presence of pi-bonding between the In and N p-orbitals, and an analogy is drawn to the frontier molecular orbitals of trimethylenemethane. The low-lying spectroscopic transition in 2, and thus its yellow color, results from mixing of the lone pair electrons on the beta-nitrogens of the hydrazido ligands with the HOMO of the InN(3) core. PMID- 12767179 TI - Iminophosphorane mediated imine metathesis. AB - The iminophosphorane Cl(3)P[double bond]NAr (1a, Ar = 2-fluorophenyl) reacts metathetically with imines at 80 degrees C to produce [double bond]NR exchange products. Compound 1a effectively catalyzes imine/imine and imine/carbodiimide cross-metathesis. The observation of [double bond]NR exchange products as well as spectroscopic evidence for the existence of diazaphosphetidine type intermediates suggests that a [2 + 2] addition/elimination mechanism is the primary pathway for substrates with N-alkyl substituents and a secondary pathway for N-aryl imines. In contrast to previously studied carbodiimide systems, the resting state of the catalyst is the iminophosphorane and not the diazaphosphetidine. For N-aryl imines, Lewis-acid catalysis appears to be the dominant mechanism, not addition/elimination. For N-alkyl imines, a decomposition pathway, involving HCl elimination from a phosphorus intermediate, is competitive in some cases. PMID- 12767181 TI - Iron and cobalt ethylene polymerization catalysts: variations on the central donor. AB - Three classes of ligands, designed to explore the effect of variations on the central pyridine donor core in bis(imino)pyridine iron and cobalt ethylene polymerization catalysts of the general formula [LMCl(2)] (M = Fe or Co), have been prepared. The first class comprises six-membered N-heterocycles (pyrimidine and triazine) and the second class five-membered heterocycles (furan and thiophene) as the central donor core. In the third class of ligands, the imine donor arm has been extended by one carbon to give anionic tridentate ligands based on carbazole and neutral analogues based on dibenzofuran and dibenzothiophene. The coordination behavior of these ligands upon reaction with FeCl(2) or CoCl(2) has been investigated, whereby only in the case of the neutral pyrimidine or the anionic carbazolide unit as the central donor core have stable complexes been obtained. Ethylene polymerization results are compared with the parent bis(imino)pyridine iron and cobalt catalyst systems. PMID- 12767180 TI - Highly sensitive luminescent metal-complex receptors for anions through charge assisted amide hydrogen bonding. AB - Two structurally simple and easily synthesized luminescent anion receptors featured with an amide-type anion binding site and rhenium(I) tricarbonyl pyridine signaling units have been developed, and they display outstanding sensitivity and selectivity toward a variety of anionic species. These complexes are highly emissive in solution. Upon anion binding, the emission intensity was significantly quenched. The sensitivities of these complexes are so high that the emission intensity can be effectively quenched by as much as 10% even in the presence of only 10(-8) M cyanide or fluoride anions. The ability of formation of intramolecular hydrogen bonding between the amide protons and central pyridine is believed to be responsible for the observed high selectivity. PMID- 12767182 TI - Coordination properties of a diarylaza crown ether appended with a luminescent [Ru(bipy)3]2+ unit. AB - The [Ru(bipy)(2)(1)](PF(6))(2) (bipy refers to 2,2'-bipyridine) complex, comprising a ruthenium(II) tris(2,2'-bipyridine) luminophore covalently linked to a di[(o-triethyleneglycoxy)phenyl]amine crown ether 1, has been synthesized and fully characterized. The photophysical properties of this metal complex have been examined in solution at ambient temperature. Luminescence from the metal complex is enhanced significantly in the presence of various adventitious cations, including protons. In particular, Li(+) cations bind to the crown ether, as evidenced by (1)H NMR and luminescence spectroscopy. Cation binding serves to decrease the rate of reductive quenching of the triplet state of the metal complex, thereby increasing the extent of luminescence. The solution-phase conformation of [Ru(bipy)(2)(1)](PF(6))(2), with and without encapsulated Li(+), has been examined by 2-D NMR and by molecular dynamics simulations. PMID- 12767184 TI - Eilatin as a bridging ligand in ruthenium(II) complexes: synthesis, crystal structures, absorption spectra, and electrochemical properties. AB - The potential of the heptacyclic aromatic alkaloid eilatin (1), that features two nonequivalent binding sites, to serve as a bridging ligand is reported. The nonequivalency of the binding sites allowed the selective synthesis of both mono- and dinuclear complexes. The mononuclear Ru(II) complexes [Ru(dmbpy)(2)(eilatin)](2+) (2) and [Ru(tmbpy)(2)(eilatin)](2+) (3) in which eilatin selectively binds "head-on" were synthesized and employed as building blocks in the synthesis of the dinuclear complexes [[Ru(dmbpy)(2)](2)(mu eilatin)](4+) (4) and [[Ru(tmbpy)(2)](2)(mu-eilatin)](4+) (5). Complete structure elucidation of the complexes in solution was accomplished by 1D and 2D NMR techniques. The X-ray structures of the mononuclear complex 3 and of the two dinuclear complexes 4 and 5 were solved, and absorption spectra and electrochemical properties of the complexes were explored. Both dinuclear complexes formed as racemic mixtures in a 3:1 diastereoisomeric ratio, the major isomer being the heterochiral one (Delta Lambda/Lambda Delta) as revealed by crystallography. The mononuclear complexes feature an exceptionally low energy MLCT band around 600 nm that shifted to over 700 nm upon the binding of the second Ru(II) center. The mononuclear complexes show one reversible oxidation and several reversible reduction waves, the first two reductions being substantially anodically shifted in comparison with [Ru(bpy)(3)](2+), attributed to the reduction of eilatin, and consistent with its low lying pi* orbital. The dinuclear complexes follow the same reduction trend, exhibiting several reversible reduction waves, and two reversible well-resolved metal centered oxidations due to the nonequivalent binding sites and to a significant metal metal interaction mediated by the bridging eilatin. PMID- 12767183 TI - Acid-base and metal ion binding properties of guanylyl(3'-->5')guanosine (GpG-) and 2'-deoxyguanylyl(3'-->5')-2'-deoxyguanosine [d(GpG)-] in aqueous solution. AB - The acidity constants of guanylyl(3'-->5')guanosine (GpG(-)) and 2' deoxyguanylyl(3'-->5')-2'-deoxyguanosine [d(GpG)(-)] for the deprotonation of their (N1)H sites were measured by potentiometric pH titrations in aqueous solution (25 degrees C; I = 0.1 M, NaNO(3)). The same method was used for the determination of the stability constants of the 1:1 complexes formed between Mg(2+), Ni(2+), or Cd(2+) (= M(2+)) and (GG-H)(2-), and in the case of Mg(2+) also of (GG-2H)(3-), where GG(-) = GpG(-) or d(GpG)(-). The stability constants of the M(GG)(+) complexes were estimated. The acidity constants of the H(dGuo)(+) and dGuo species (dGuo = 2'-deoxyguanosine) and the stability constants of the corresponding M(dGuo)(2+) and M(dGuo-H)(+) complexes were also measured. Comparison of these and related data allows the conclusion that N7 of the 5'G unit in GG(-) is somewhat more basic than the one in the 3'G moiety; the same holds for the (N1)(-) sites. On the basis of comparisons with the stability constants measured for the dGuo complexes, it is concluded that M(2+) binding of the GG dinucleoside monophosphates occurs predominantly in a mono-site fashion, meaning that macrochelate formation is not very pronounced. Indeed, it was a surprise to find that the stabilities of the complexes of dGuo or (dGuo-H)(-) and the corresponding ones derived from GG(-) are so similar. Consequently, it is suggested that in the M(GG)(+) and M(GG-H) complexes the metal ion is mainly located at N7 of the 5'G unit since this is the more basic site allowing also an outer-sphere interaction with the C6 carbonyl oxygen and because this coordination mode is also favorable for an electrostatic interaction with the negatively charged phosphodiester bridge. It is further suggested that Mg(2+) binding (which is rather weak compared to that of Ni(2+) and Cd(2+)) occurs mainly in an outer-sphere mode, and on the basis of the so-called Stability Ruler it is concluded that the binding properties of Zn(2+) to the GG species are similar to those of Ni(2+) and Cd(2+). PMID- 12767185 TI - Antiferromagnetic three-dimensional order induced by carboxylate bridges in a two dimensional network of [Cu3(dcp)2(H2O)4] trimers. AB - A new Cu(II) complex, [Cu(3)(dcp)(2)(H(2)O)(4)](n), with the ligand 3,5 pyrazoledicarboxylic acid monohydrate (H(3)dcp) has been prepared by hydrothermal synthesis, and it crystallizes in the monoclinic space group P2(1)/c with a = 11.633(2) A, b = 9.6005(14) A, c = 6.9230(17) A, beta = 106.01(2) degrees, and Z = 2. In the solid state structure of [Cu(3)(dcp)(2)(H(2)O)(4)](n), trinuclear [Cu(3)(dcp)(2)(H(2)O)(4)] repeating units in which two dcp(3-) ligands chelate the three Cu(II) ions with the central Cu(II) ion, Cu(1) (on an inversion center), link to form infinite 2D sheets via syn-anti equatorial-equatorial carboxylate bridges between Cu(2) atoms in adjacent trimers. These layers are further linked by syn-anti axial-equatorial carboxylate bridging between Cu(1) atoms in adjacent sheets resulting in the formation of a crystallographic 3D network. A detailed analysis of the magnetic properties of [Cu(3)(dcp)(2)(H(2)O)(4)](n) reveals that the dcp(3-) ligand acts to link Cu(II) centers in three different ways with coupling constants orders of magnitude apart in value. In the high temperature region above 50 K, the dominant interaction is strongly antiferromagnetic (J/k(B) = -32 K) within the trimer units mediated by the pyrazolate bridges. Below 20 K, the trimer motif can be modeled as an S = 1/2 unit. These units are coupled to their neighbors by a ferromagnetic interaction mediated by the syn-anti equatorial-equatorial carboxylate bridge. This interaction has been estimated at J(2D)/k(B) = +2.8 K on the basis of a 2D square lattice Heisenberg model. Finally, below 3.2 K a weak antiferromagnetic coupling (J(3D)/k(B) = -0.1 K) which is mediated by the syn-anti axial-equatorial carboxylate bridges between the 2D layers becomes relevant to describe the magnetic (T, H) phase diagram of this material. PMID- 12767186 TI - Self-assembly of a series of novel metal-organic compounds containing ferrocenecarboxylate components. AB - Using FcCOONa (Fc = (eta(5)-C(5)H(5))Fe(eta(5)-C(5)H(4))) as starting material, we obtained an unprecedented metal-organic coordination polymer containing ferrocenecarboxylate components [[Pb(2)(FcCOO)(eta(2)-FcCOO)(mu(2)-eta(2) FcCOO)(mu(3)-eta(2)-FcCOO)(CH(3)OH)].1.5CH(3)OH.H(2)O](n) (1), tetramer [Zn(4)(mu(2)-FcCOO)(6)(mu(4)-O)] (2), and coordination polymers [Pb(FcCOO)(mu(2) FcCOO)(bpe)](n) (3) (bpe = 1,2-bis(4-pyridyl)ethene), [[Zn(FcCOO)(2)(bpt)].2.5H(2)O](n) (4) (bpt = N,N'-bis(3-pyridylmethyl)thiourea), and [Zn(FcCOO)(eta(2)-FcCOO)(bbp)](n) (5) (bbp = 4,4'-trimethylene-dipyridine). Compounds 1 and 2 are formed by ferrocenecarboxylate units coordinating with Pb(II) or Zn(II). In polymer 1, ferrocenecarboxylate units have four kinds of coordinate modes; just these novel coordinate modes lead to the unprecedented one dimensional polymer where two kinds of rhomboids are arranged alternatively along the chain. Compound 2 is a tetramer, in which a distinct connectivity of the six ferrocene units is established through the four Zn atoms. Compounds 3-5 are obtained by organic ligands bridging Pb(II) or Zn(II), leading to a new type of metal-organic coordination polymer. PMID- 12767187 TI - Structure and magnetic properties of the two-dimensional ferrimagnet (NEt4)[[Mn(salen)]2Fe(CN)6]: investigation of magnetic anisotropy on a single crystal. AB - The title compound, (NEt(4))[[Mn(salen)](2)Fe(CN)(6)] (1), was synthesized via a 1:1 reaction of [Mn(salen)(H(2)O)]ClO(4) with (NEt(4))(3)[Fe(CN)(6)] in a methanol/ethanol medium (NEt(4)(+) = tetraethylammonium cation, salen(2)(-) = N,N'-ethylenebis(salicylidene)iminate). The two-dimensional layered structure of 1 was revealed by X-ray crystallographic analysis: 1 crystallizes in monoclinic space group P2(1)/c with cell dimensions of a = 12.3660(8) A, b = 15.311(1) A, c = 12.918(1) A, beta = 110.971(4) degrees, Z = 2 and is isostructural to the previously synthesized compound, (NEt(4))[[Mn(5-Clsalen)](2)Fe(CN)(6)] (5 Clsalen(2-) = N,N'-ethylenebis(5-chlorosalicylidene)iminate; Miyasaka, H.; Matsumoto, N.; Re, N.; Gallo, E.; Floriani, C. Inorg. Chem. 1997, 36, 670). The Mn ion is surrounded by an equatorial salen quadridentate ligand and two axial nitrogen atoms from the [Fe(CN)(6)](3-) unit, the four Fe[bond]CN groups of which coordinate to the Mn ions of [Mn(salen)](+) units, forming a two-dimensional network having [[bond]Mn[bond]NC[bond]Fe[bond]CN[bond]](4) cyclic repeating units. The network is spread over the bc-plane of the unit cell, and the layers are stacked along the a-axis. The countercation NEt(4)(+) is located between the layers. Compound 1 is a ferrimagnet with T(c) = 7.7 K and exhibits hysteresis with a remnant magnetization of 13.44 cm(3).mol(-1) (M/N mu(B) = 2.4) at zero field and a coercivity of 1000 Oe when the powder sample was measured at 1.9 K. Magnetic measurements of a direction-arranged single crystal were also carried out. The orientation of the crystallographic axes of a selected single crystal was determined by X-ray analysis, and magnetization was measured when an external field was applied in the a*, b, and c directions. The magnetization in the a* direction increased more easily than those in the b and c directions below the critical temperature. No hysteresis was observed only for the measurement in the a* direction, indicating the presence of strong structural anisotropy with potential anisotropy on Mn(III) ions. PMID- 12767189 TI - Donor-functionalized lanthanide terphenyl complexes: Synthesis and structural characterization of 2,6-di(o-anisol)phenyl compounds of ytterbium, yttrium, and samarium. AB - The syntheses and molecular structures of a number of 2,6-di(o-anisol)phenyl ([double bond]Danip-) -based bis(amide) and bis(alkoxide) compounds of ytterbium, yttrium, and samarium are reported. Additionally, NMR spectroscopic data are reported for the analogous diamagnetic yttrium compounds. Salt metathesis reaction of equimolar amounts of DanipLi and YbCl(3) in tetrahydrofuran at room temperature followed by addition of 2 equiv of KN(SiMe(3))(2) or KN(SiHMe(2))(2) produces DanipYb[N(SiMe(3))(2)](2) (1) and DanipYb[N(SiHMe(2))(2)](2) (2), respectively. The analogous reaction using SmCl(3) and KN(SiHMe(2))(2) produces DanipSm[N(SiHMe(2))(2)](2) (3). Reaction of DanipLi and YbCl(3) in tetrahydrofuran at room temperature followed by addition of 2 equiv of KO(2,6 diisopropylphenyl) produces DanipYb[O(2,6-diisopropylphenyl)](2) (4). Our attempts to also prepare the yttrium analogue of complex 4 yielded single crystalline material of the tetrahydrofuran adduct DanipY(THF)[O(2,6 diisopropylphenyl)](2) (5). The molecular structures of the complexes 1-4 feature five-coordinate metal atoms and coordination polyhedra which can be described as distorted square-pyramidal rather than trigonal-bipyramidal, with the ipso carbon atom occupying the apical position. On the other hand, the molecular structure of the tetrahydrofuran-solvated yttrium Danip arylalkoxide compound 5 features a six coordinate metal atom in a distorted trigonal-prismatic coordination environment. In all cases the Danip ligand system adopts the chiral (racemic) d,l form. PMID- 12767188 TI - Reactivity of the organometallic fac-[(CO)3ReI(H2O)3]+ aquaion. Kinetic and thermodynamic properties of H2O substitution. AB - The water exchange process on [(CO)(3)Re(H(2)O)(3)](+) (1) was kinetically investigated by (17)O NMR. The acidity dependence of the observed rate constant k(obs) was analyzed with a two pathways model in which k(ex) (k(ex)(298) = (6.3 +/- 0.1) x 10(-3) s(-1)) and k(OH) (k(OH)(298)= 27 +/- 1 s(-1)) denote the water exchange rate constants on 1 and on the monohydroxo species [(CO)(3)Re(I)(H(2)O)(2)(OH)], respectively. The kinetic contribution of the basic form was proved to be significant only at [H(+)] < 3 x 10(-3) M. Above this limiting [H(+)] concentration, kinetic investigations can be unambiguously conducted on the triaqua cation (1). The variable temperature study has led to the determination of the activation parameters Delta H(++)(ex) = 90 +/- 3 kJ mol( 1), Delta S(++)(ex) = +14 +/- 10 J K(-1) mol(-1), the latter being indicative of a dissociative activation mode for the water exchange process. To support this assumption, water substitution reaction on 1 has been followed by (17)O/(1)H/(13)C/(19)F NMR with ligands of various nucleophilicities (TFA, Br(-), CH(3)CN, Hbipy(+), Hphen(+), DMS, TU). With unidentate ligands, except Br(-), the mono-, bi-, and tricomplexes were formed by water substitution. With bidentate ligands, bipy and phen, the chelate complexes [(CO)(3)Re(H(2)O)(bipy)]CF(3)SO(3) (2) and [(CO)(3)Re(H(2)O)(phen)](NO(3))(0.5)(CF(3)SO(3))(0.5).H(2)O (3) were isolated and X-ray characterized. For each ligand, the calculated interchange rate constants k'(i) (2.9 x 10(-3) (TFA) < k'(I) < 41.5 x 10(-3) (TU) s(-1)) were found in the same order as the water exchange rate constant k(ex), the S-donor ligands being slightly more reactive. This result is indicative of I(d) mechanism for water exchange and complex formation, since larger variations of k'(i) are expected for an associatively activated mechanism. PMID- 12767190 TI - Additional steps toward molecular scale wires: Further study of Ni(5)(10/11+) chains embraced by polypyridylamide ligands. AB - This paper presents two advances in the development of the chemistry of extended metal atom chains (EMACs) that employ di(2-pyridyl)amide (dpa) and its higher homologues (loosely called polypyridylamides). As EMACs employing these ligands are extended to greater lengths, low solubility becomes an increasingly difficult problem. Also, increased stability would be desirable. We have employed a method, which is designed to be applicable to chains of any length, to introduce stabilizing substituents (ethyl groups) on some of the pyridyl rings. We illustrate this here by the synthesis and characterization of the pentanickel complexes Ni(5)(etpda)(4)Cl(2).6CHCl(3) and [Ni(5)(etpda)(4)](PF(6))(3).4Me(2)CO, etpda = the anion of N,N'-bis(4-ethylpyridyl)-2,6-diaminopyridine. As we had previously predicted, on the basis of the behavior of Ni(3)(dpa)(4)Cl(2) and [Ni(3)(dpa)(4)](PF(6))(3), oxidation causes marked changes in structure and magnetic behavior indicative of a change of electronic structure that would cause an insulator-conductor transformation. We now demonstrate that this is what occurs not only in the previously known Ni(5) compounds but in the new ethyl substituted ones. PMID- 12767191 TI - Aggregate manganese Schiff base moieties by terephthalate or acetate: dinuclear manganese and trinuclear mixed metal Mn2/Na complexes. AB - A reaction system consisting of terephthalic acid, NaOH, inorganic Mn(II) or Mn(III) salt, and salicylidene alkylimine resulted in dinuclear manganese complexes (salpn)(2)Mn(2)(mu-phth)(CH(3)OH)(2) (1, salpn = N,N'-1,3-propylene bis(salicylideneiminato); phth = terephthalate dianion), (salen)(2)Mn(2)(mu phth)(CH(3)OH)(2) (2, salen = N,N'-ethylene-bis(salicylideneiminato)), (salen)(2)Mn(2)(mu-phth)(CH(3)OH)(H(2)O) (3), and (salen)(2)Mn(2)(mu-phth) (4), while the absence of NaOH in the reaction led to a mononuclear Mn complex (salph)Mn(CH(3)OH)(NO(3)) (5, salph = N,N'-1,2-phenylene bis(salicylideneiminato)). In addition, a trinuclear mixed metal complex H[Mn(2)Na(salpn)(2)(mu-OAc)(2)(H(2)O)(2)](OAc)(2) (6) was obtained from the reaction system by using maleic acid instead of terephthalic acid. Five coordinate Mn ions were found in 4 giving rise to an intermolecular interaction and constructing a one-dimensional linear structure. Antiferromagnetic exchange interactions were observed for 1-3, and a total ferromagnetic exchange of 4 was considered to stem from intermolecular magnetic coupling. (1)H NMR signals of phenolate ring and alkylene (or phenylene) backbone of the diamine are similar to those reported in the literature, and the phth protons are at -2.3 to -10.1 ppm. Studies on structure, bond valence sum analysis, and magnetic properties indicate the oxidation states of the Mn ions in 6 to be +3, which are also indicated by ESR spectra in dual mode. Ferromagnetic exchange interaction between the Mn(III) sites was observed with J = 1.74 cm(-1). A quasireversible redox pair at -0.29V/ 0.12V has been assigned to the redox of Mn(2)(III)/Mn(III)Mn(II), implying the intactness of the complex backbone in solution. PMID- 12767192 TI - Synthesis, structural investigation, and solid-state properties of iodine-doped zirconium diphthalocyanine, [ZrPc2]I3.I2. AB - Crystals of iodine-doped zirconium(IV) diphthalocyanine, [ZrPc(2)]I(3).I(2) (where Pc = C(32)H(16)N(8)), were grown directly in the reaction of pure zirconium powder with phthalonitrile under a stream of iodine at 260 degrees C. [ZrPc(2)]I(3).I(2) crystallizes in the space group P2(1)/m (No. 11) of the monoclinic system with lattice parameters of a = 6.735(1), b = 25.023(5), and c = 17.440(3) A, beta = 99.43(3) degrees, and Z = 2. The crystals of [ZrPc(2)]I(3).I(2) are built up from two pseudo-monodimensional aggregates: one electron-oxidized [ZrPc(2)](+) units; weak interacting triiodide I(3)(-) ions with neutral diiodine molecules. The I(3)(-) ions and neutral I(2) molecules in the crystal of [ZrPc(2)]I(3).I(2) have been also detected by Raman spectroscopy. The [ZrPc(2)](+) units form stacks along the a axis, while the polymeric...I(3)( )...I(2)...I(3)(-)...I(2)(-)... zigzag chains are located in the crystal along the b axis, so both pseudo-monodimensional aggregates are perpendicular to each other. This arrangement is different from that found in the tetragonal crystals of [ZrPc(2)](I(3))(2/3) in which both monodimensional aggregates, i.e., the stacks of partially oxidized [ZrPc(2)](2/3+) units and chains of symmetric triiodide ions, are parallel. EPR experiment together with the X-ray single crystal analysis clearly shown that oxidation of the diamagnetic ZrPc(2) complex by iodine is ligand centered and homogeneously affecting both phthalocyaninato rings of ZrPc(2); thus, the formal oxidation state of both Pc rings in [ZrPc(2)]I(3).I(2) is nonintegral (-1.5). The UV-vis spectrum of [ZrPc(2)] I(3).I(2) is very similar to the spectrum of unoxidized ZrPc(2) complex in the B Soret and Q spectral region. However, in the spectrum of [ZrPc(2)] I(3).I(2) one additional band at approximately 502 nm is observed, which indicates the existence of the one-electron-oxidized phthalocyaninato(-) radical ligand and is assigned to the electronic transition from a deeper level to the half-occupied HOMO level. The single-crystal electrical conductivity data show anisotropy and nonmetallic character in conductivity (d sigma/dT > 0). The charge transport mainly proceeds along the pseudo-monodimensional stacks of [ZrPc(2)](+) units. The relatively high conductivity along the stacks of one-electron-oxidized [ZrPc(2)](+) units results from the staggering orientation of Pc rings (rotation angle 45.0(2) degrees ) that leads to the short inter-ring C(alpha)(pyrrole)[bond]C(alpha)(pyrrole) contacts (2.839(3)-3.024(3) A). These C(alpha)-pyrrole atoms make appreciable contribution to the partially occupied pi molecular orbital of Pc macrocycle and the greatest overlap of the HOMO orbitals that form the conduction band of partially oxidized molecular crystals. PMID- 12767193 TI - Formation and photoinduced electron ejection of amminedicyanocuprate(I) and cyano bridged trinuclear copper(I) complexes in aqueous ammonia solution. AB - UV absorption spectral evidence confirms that a mixed-ligand complex, Cu(CN)(2)(NH(3))(-), is formed in an aqueous solution of KCu(CN)(2) and ammonia. The stepwise stability constant for the reaction, Cu(CN)(2)(-) + NH(3) = Cu(CN)(2)(NH(3))(-), is 2.80 +/- 0.40 in 1 M ionic strength, NaClO(4) medium at 25 degrees C. This amminedicyanocuprate(I) ion readily combines in aqueous solution in a 1:2 and 2:1 molar ratios with Cu(NH(3))(2)(+) to form two trinuclear ionic species, presumably with cyano bridges, with the suggested formulas of Cu(3)(CN)(2)(NH(3))(5)(+) and Cu(3)(CN)(4)(NH(3))(3)(-). The resolved UV absorption spectra of the monomer and two trimers have been determined and exhibit strong bands, presumably metal-ligand charge transfer in nature, in the 200-250-nm region. When solutions of all three complexes absorb a pulse of 266-nm laser light, they photoeject hydrated electrons monophotonically, with quantum yields of 0.41 +/- 0.02, 0.53 +/- 0.02, and 0.50 +/- 0.01 for the monomer, cationic trimer, and anionic trimer, respectively, suggesting that absorption in the charge-transfer-to-solvent bands of these complexes results in an efficient electron ejection process that is enhanced by the existence of a polynuclear structure with cyano bridges. No room-temperature luminescence is observed for these complexes. PMID- 12767194 TI - A resonance Raman, surface-enhanced resonance Raman, IR, and ab initio vibrational spectroscopic study of nickel(II) tetraazaannulene complexes. AB - The IR and resonance Raman spectra of the nickel(II) complexes of dibenzo[b,i][1,4,8,11]tetraaza[14]annulene (TAA) and 5,7,12,14 tetramethyldibenzo[b,i][1,4,8,11]tetraaza[14]annulene (TMTAA) have been measured and compared with ab initio calculations of the vibrational wavenumbers at the B3 LYP level using the LanL2DZ basis set. An excellent fit is found between the experimental and calculated data, enabling precise vibrational assignments to be made. Surface-enhanced resonance Raman spectra were obtained following adsorption on Ag electrodes, with potentials in the range -0.1 to -1.1 V vs Ag/AgCl. There is evidence for contributions from both the electromagnetic and charge transfer (CT) surface enhancement mechanisms. The data indicate that variations in band intensities with electrode potential can be interpreted in terms of the CT mechanism. PMID- 12767195 TI - Electronic effects in 12-metallacrown-3 complexes. A theoretical and experimental study. AB - The Mulliken charges of the 12-metallacrown-3 complex [(C(6)H(6))Ru(C(5)H(3)NO(2))](3) were determined by a single point analysis at the HF level. For comparison, a Mulliken population analysis was carried out for the organic analogue 12-crown-3. The partial negative charges on the O-donor atoms of the metallamacrocycle were found to be larger than those on the O-donor atoms of 12-crown-3. The 12-metallacrown-3 complex [(cymene)Ru(C(5)H(2)ClNO(2))](3) with chloro-substituents in position 5 of the pyridonate ligand was synthesized to determine the effect of electron withdrawing groups on the structure and the host-guest properties of the receptor. The chloro substituents were found to have only a small influence on the structures, but they reduce the binding affinity for LiCl and NaCl by approximately 2 orders of magnitude. PMID- 12767197 TI - Diphenyldichlorophosphonium trichloride-chlorine solvate 1:1, [PPh2Cl2]+Cl(3)( ).Cl2: an ionic form of diphenyltrichlorophosphorane. Crystal structures of [PPh2Cl2]+Cl(3)(-).Cl2 and [(PPh2Cl2)+]2[InCl5](2-). AB - An ionic form of diphenyltrichlorophosphorane, namely, diphenyldichlorophosphonium trichloride isolated as a dichlorine solvate (1), was obtained by treating PPh(2)Cl(3) with excess chlorine. The identity of this species was established by single-crystal X-ray analysis and (31)P, (1)H, and (35)Cl NMR and Raman spectra. Bis(diphenyldichlorophosphonium) pentachloroindate (2) was obtained by the reaction of diphenyltrichlorophosphorane with indium trichloride in dichloromethane for comparison purposes. Its identity was determined by (31)P NMR spectra and single-crystal X-ray analysis. PMID- 12767196 TI - Investigations into the interactions between DNA and conformationally constrained pyridylamineplatinum(II) analogues of AMD473. AB - The syntheses of [PtCl(2)(amp)] (amp = 2-pyridylmethylamine) and enantiomerically pure [PtCl(2)(R-pea)] and [PtCl(2)(S-pea)] (pea = 1-(2-pyridyl)ethylamine) and the crystal structure of [PtCl(2)(R-pea)] are reported. The reactions of [PtCl(2)(amp)] and of the enantiomers of [PtCl(2)(pea)] with d(GpG) and with a 52 base-pair oligonucleotide were investigated. Each of the reactions with d(GpG) resulted in the formation of three platinated bifunctional d(GpG) species in a ratio of 1:2:1. These species were shown to be a pair of isomers, one of which exists as a pair of slowly interconverting rotamers that can be separated by HPLC but reequilibrate after 5 days at 37 degrees C. The pyridyl moieties of the pyridylalkylamine ligands are constrained to lie in the coordination plane, and as a consequence, the rotation about the Pt-N7 bond of the adjacent guanine is highly restricted. 2D NMR investigations were carried out on the isomer of [Ptd(GpG)(amp)] that did not form separable rotamers and identified it as the isomer having the pyridine adjacent to the 5'-guanine of the d(GpG). The reaction of each of the three [PtCl(2)(py-R)] complexes (py-R = amp or pea) with a 52-base pair oligonucleotide resulted in the formation of the same three bifunctional d(GpG) adducts in approximately the same ratios as the reactions with d(GpG), indicating that negligible stereoselectivity results from interactions between the complexes and duplex DNA. PMID- 12767198 TI - Enhancing the stability of trinickel molecular wires and switches: Ni(3)(6+)/Ni(3)(7+). AB - This paper describes in detail four new compounds that contain extended metal atom chains (EMACs) of three nickel atoms wrapped by either di(2-pyridyl)amide (dpa) or the new homologous ligand with an ethyl group at the para position of each pyridyl group, depa, and compares them to the precursor Ni(3)(dpa)(4)Cl(2) (1) and the oxidized and rather unstable Ni(3)(dpa)(4)(PF(6))(3) (2). The new molecules are Ni(3)(depa)(4)Cl(2) (3), Ni(3)(depa)(4)(PF(6))(3) (4), [Ni(3)(dpa)(4)(CH(3)CN)(2)](PF(6))(2) (5), and [Ni(3)(depa)(4)(CH(3)CN)(2)](PF(6))(2) (6). These compounds are fully described as to preparation, elemental composition, structure, infrared spectra, (1)H NMR spectra (where possible), electrochemistry, magnetic susceptibility, and an EPR spectrum for 4. The effects of (a) introducing the ethyl substituents on the ligands, (b) replacing axial anions by neutral axial ligands, and (c) oxidizing the Ni(3) chains are reported and discussed. The point of major interest is how oxidation profoundly alters the electronic structure of the EMAC. PMID- 12767199 TI - Pop-the-cork strategy in synthetic utilization of imines: stabilization by complexation and activation via liberation of the ligated species. AB - Treatment of trans-[PtCl(4)(RCN)(2)] (R = Me, Et) with ethanol allowed the isolation of trans-[PtCl(4)[E-NH[double bond]C(R)OEt](2)]. The latter were reduced selectively, by the ylide Ph(3)P[double bond]CHCO(2)Me, to trans [PtCl(2)[E-NH[double bond]C(R)OEt](2)]. The complexed imino esters NH[double bond]C(R)OEt were liberated from the platinum(II) complexes by reaction with 2 equiv of 1,2-bis(diphenylphosphino)ethane (dppe) in chloroform; the cationic complex [Pt(dppe)(2)]Cl(2) precipitates almost quantitatively from the reaction mixture and can be easily separated by filtration to give a solution of NH[double bond]C(R)OEt with a known concentration of the imino ester. The imino esters efficiently couple with the coordinated nitriles in trans-[PtCl(4)(EtCN)(2)] to give, as the dominant product, [PtCl(4)[NH[double bond]C(Et)N[double bond]C(R)OEt](2)] containing a previously unknown linkage, i.e., ligated N-(1 imino-propyl)-alkylimidic acid ethyl esters. In addition to [PtCl(4)[NH[double bond]C(Et)N[double bond]C(Et)OEt](2)], another compound was generated as the minor product, i.e., [PtCl(4)(EtCN)[NH[double bond]C(Et)N[double bond]C(Et)OEt]], which was reduced to [PtCl(2)(EtCN)[NH[double bond]C(Et)N[double bond]C(Et)OEt]], and this complex was characterized by X-ray single-crystal diffraction. The platinum(IV) complexes [PtCl(4)[NH[double bond]C(Et)N[double bond]C(R)OEt](2)] are unstable toward hydrolysis and give EtOH and the acylamidine complexes trans [PtCl(4)[Z-NH[double bond]C(Et)NHC(R)[double bond]O](2)], where the coordination to the Pt center results in the predominant stabilization of the imino tautomer NH[double bond]C(Et)NHC(R)[double bond]O over the other form, i.e., NH(2)C(Et)[double bond]NC(R)[double bond]O, which is the major one for free acylamidines. The structures of trans-[PtCl(4)[Z-NH[double bond]C(Et)NHC(R)[double bond]O](2)] (R = Me, Et) were determined by X-ray studies. The complexes [PtCl(4)[NH[double bond]C(Et)N[double bond]C(R)OEt](2)] were reduced to the appropriate platinum(II) compounds [PtCl(2)[NH[double bond]C(Et)N[double bond]C(R)OEt](2)], which, similarly to the appropriate Pt(IV) compounds, rapidly hydrolyze to yield the acylamidine complexes [PtCl(2)[NH[double bond]C(Et)NHC(R)[double bond]O](2)] and EtOH. The latter acylamidine compounds were also prepared by an alternative route upon reduction of the corresponding platinum(IV) complexes. Besides the first observation of the platinum(IV)-mediated nitrile-imine ester integration, this work demonstrates that the application of metal complexes gives new opportunities for the generation of a great variety of imines (sometimes unreachable in pure organic chemistry) in metal-mediated conversions of organonitriles, the "storage" of imino species in the complexed form, and their synthetic utilization after liberation. PMID- 12767200 TI - A monovacant heteropolytungstate thioderivative: Synthesis and characterization of [(PW11O39)2(H4Mo4S4O6)](10-) and related isomers. AB - [(PW(11)O(39))(2)(Mo(4)S(4)O(4)(OH(2))(2))](10-) anions were obtained through the stereospecific addition of the [Mo(2)S(2)O(2)](2+) oxothiocation to the monovacant alpha-[PW(11)O(39)](7-) anion. K(10)[(PW(11)O(39))(2)(Mo(4)S(4)O(4)(OH(2))(2))].25H(2)O has been isolated as crystals and characterized by X-ray diffraction. The structure revealed a "sandwich-like" dimer of two alpha-[PW(11)O(39)](7-) subunits assembled by the noteworthy central cluster [H(4)Mo(4)S(4)O(6)]. The crystallization of the crude product produces an isomerically pure compound, which was characterized by (31)P and (183)W NMR. IR data were also supplied. In solution, the compound isomerizes, giving a second diastereoisomer. A kinetic experiment, carried out by (31)P NMR, allowed the conditions of the thermodynamic equilibrium to be determined. A structural relationship between the two isomers is proposed, fully consistent with NMR data. Cisoid and transoid isomers result in the relative disposition of each [PW(11)O(39)](7-) subunit, either staggered or eclipsed. An investigation of the formation of the [Mo(2)O(2)S(2)](2+) unit from the polycondensed cyclic precursor [Mo(10)S(10)O(10)(OH)(10)(H(2)O)(5)] and the aggregation process resulting in the oxothio [(PW(11)O(39))(2)(Mo(4)S(4)O(4)(OH(2))(2))](10-) compound has been undertaken. The studies were monitored by (31)P NMR and UV-vis spectroscopies. The reaction is quantitative in nearly stoichiometric conditions. PMID- 12767201 TI - Metal-bis[poly(pyrazolyl)borate] complexes. Electrochemical, magnetic, and spectroscopic properties and coupled electron-transfer and spin-exchange reactions. AB - Electrochemical, magnetic, and spectroscopic properties are reported for homoleptic divalent (M = Mn, Fe, Co, Ni, Ru) and trivalent (M = Cr, Mn, Fe, Co) metal-bis[poly(pyrazolyl)borate] complexes, [M(pzb)(2)](+/0), where pzb(-) = hydrotris(pyrazolyl)borate (Tp), hydrotris(3,5-dimethylpyrazolyl)borate (Tp), or tetrakis(pyrazolyl)borate (pzTp). Ligand field strengths in metal-pzb complexes increase as Tp < Tp < pzTp, which reflects the importance of steric rather than electronic effects on spectroscopic properties. However, metal-centered redox potentials become more negative as pzTp < Tp < Tp, which follows the electron donating ability of the ligands. Co(III)/Co(II) and Mn(III)/Mn(II) electrode reactions are accompanied by a change in metal atom spin-state; i.e., (S = 0) [Co(pzb)(2)](+) + e(-) <==> (S = 3/2) [Co(pzb)(2)] and (S = 1) [Mn(pzb)(2)](+) + e(-) <==> (S = 5/2) [Mn(pzb)(2)]. Apparent heterogeneous electron-transfer rate constants derived from sweep-rate dependent cyclic voltammetric peak potential separations in 1,2-dichloroethane are small and decrease as pzTp > Tp > Tp for the Co(III)/Co(II) couples. Slow electron transfer is characteristic of coupled electron transfer and spin exchange. [M(Tp)(2)](+/0) redox potentials relative to values for other homoleptic MN(6)(3+/2+) couples change as M varies from Cr to Ni. For early members of the series, [M(Tp)(2)](+/0) potentials nearly equal those of complexes with aliphatic N-donor ligands (e.g., triazacyclononane, sarcophagine). However, [M(Tp)(2)](+/0) potentials approach those of [M(bpy)(3)](3+/2+) for later members of the series. The variation suggests a change in the nature of the metal-pzb interaction upon crossing the first transition row. PMID- 12767202 TI - Bis(trifluoromethyl)phosphane and bis(pentafluorophenyl)phosphane and their pentacarbonyl tungsten complexes: improved synthesis and an experimental and density functional study. AB - The use of Bu(3)SnH and Me(3)SnH in the synthesis of HP(CF(3))(2) and HP(C(6)F(5))(2) from the corresponding bromides leads to a high-yield synthesis, which additionally provides these compounds in large quantities. The pentacarbonyl tungsten complexes [W(CO)(5)PH(CF(3))(2)] and [W(CO)(5)PH(C(6)F(5))(2)] were synthesized reacting the corresponding phosphanes with [W(CO)(5)THF] and characterized by X-ray and elemental analysis as well as multinuclear NMR and mass spectroscopy. The vibrational analyses of HP(CF(3))(2) and HP(C(6)F(5))(2) and their tungsten pentacarbonyl complexes were achieved in combination with hybrid DFT calculations. The optimized structures of [W(CO)(5)PH(CF(3))(2)] and [W(CO)(5)PH(C(6)F(5))(2)] at the B3PW91 level of theory using a LanL2DZ basis and ECP at the tungsten atom and a 6-311G(3d,p) and 6-311G(d,p) basis set for the nonmetal atoms, respectively, yield an impressively good agreement between experimental and theoretical geometric parameters. An increased pi-acidity of HP(CF(3))(2) in comparison with HP(C(6)F(5))(2) and HPPh(2) is discussed in the context of vibrational analysis, X-ray structural investigations, and theoretical calculations. PMID- 12767203 TI - Stabilization of the P(CF3)(2)(-) and P(C6F5)(2)(-) ions by coordination to pentacarbonyl tungsten: structures of [18-crown-6-K]P(CF3)2, [18-crown-6 K][W[P(CF3)2](CO)5], and [18-crown-6-K][[W(CO)5]2[mu-P(C6F5)2]].THF. AB - The stabilization of the P(CF(3))(2)(-) ion by intermediary coordination to the very weak Lewis acid acetone gives access to single crystals of [18-crown-6 K]P(CF(3))(2). The X-ray single crystal analysis exhibits nearly isolated P(CF(3))(2)(-) ions with an unusually short P-C distance of 184(1) pm, which can be explained by negative hyperconjugation and is also found by quantum chemical hybrid DFT calculation. Coordination of the P(CF(3))(2)(-) ion to pentacarbonyl tungsten has only a minor effect on electronic and geometric properties of the P(CF(3))(2) moiety, while a strong increase in thermal stability of the dissolved species is achieved. The hitherto unknown P(C(6)F(5))(2)(-) ion is stabilized by coordination to pentacarbonyl tungsten and isolated as a stable 18-crown-6 potassium salt, [18-crown-6-K][W[P(C(6)F(5))(2)](CO)(5)], which is fully characterized. The tungstate, [W[P(C(6)F(5))(2)](CO)(5)](-), decomposes slowly in solution, while coordination of the phosphorus atom to a second pentacarbonyl tungsten moiety results in an enhanced thermal stability in solution. The single crystal X-ray analysis of [18-crown-6-K][[W(CO)(5)](2)[mu-P(C(6)F(5))(2)]].THF exhibits a very tight arrangement of the two C(6)F(5) and two W(CO)(5) groups around the central phosphorus atom. NMR spectroscopic investigations of the [[W(CO)(5)](2)[mu-P(C(6)F(5))(2)]](-) ion exhibit a hindered rotation of both the C(6)F(5) and W(CO)(5) groups in solution. PMID- 12767204 TI - New one-dimensional azido-bridged manganese(II) coordination polymers exhibiting alternating ferromagnetic-antiferromagnetic interactions: structural and magnetic studies. AB - Five Mn(II)[bond]azido coordination polymers of formula [Mn(L)(N(3))(2)](n) have been synthesized and crystallographically characterized, and their magnetic properties studied, where L's are the bidentate Schiff bases obtained from the condensation of pyridine-2-carbaldehyde with aniline (1) and its derivatives p toluidine (2), m-toluidine (3), p-chloroaniline (4), and m-chloroaniline (5). All the complexes consist of the zigzag Mn(II)[bond]azido chains in which the Mn(II) ions are alternately bridged by two end-to-end (EE) and two end-on (EO) azido ligands, the cis-octahedral coordination being completed by the two nitrogen atoms of the Schiff base ligands. Compound 2 is unique in that the Mn[bond](EE N(3))(2)[bond]Mn ring adopts an unusual twist conformation with the two linear azido bridges crossing each other. By contrast, the rings in the other compounds take the usual chair conformation with the two azido bridges parallel. The double EO bridging fragments in the complexes are similar with the bridging angles (Mn[bond]N[bond]Mn) ranging from 99.6 degrees to 104.0 degrees. Magnetic analyses reveal that alternating ferro- and antiferromagnetic interactions are mediated through the alternating EO and EE azido bridges with the J(F) and J(AF) parameters in the ranges of 4.1-8.0 and -11.8 to -15.4 cm(-1), respectively. Finally, the magnetostructural correlations are investigated. The present complexes follow the general trend that the ferromagnetic interaction through the double EO bridge increases with the Mn[bond]N[bond]Mn bridging angle, while the antiferromagnetic interaction through the double EE bridge is dependent on the distortion of the Mn[bond](N(3))(2)[bond]Mn ring from planarity toward the chair conformation and the Mn[bond]N[bond]N angle. PMID- 12767205 TI - Insertion reactions into Pd[bond]O and Pd[bond]N bonds: preparation of alkoxycarbonyl, carbonato, carbamato, thiocarbamate, and thioureide complexes of palladium(II). AB - Mononuclear palladium hydroxo complexes of the type [Pd(N[bond]N)(C(6)F(5))(OH)] [(N[bond]N = 2,2'-bipyridine (bipy), 4,4'-dimethyl-2,2'-bipyridine (Me(2)bipy), 1,10-phenanthroline (phen), or N,N,N',N'-tetramethylethylenediamine (tmeda)] have been prepared by reaction of [Pd(N[bond]N)(C(6)F(5))(acetone)]ClO(4) with KOH in methanol. These hydroxo complexes react, in methanol, with CO (1 atm, room temperature) to yield the corresponding methoxycarbonyl complexes [Pd(N[bond]N)(C(6)F(5))(CO(2)Me)]. Similar alkoxycarbonyl complexes [Pd(N[bond]N)(C(6)F(5))(CO(2)R)] (N[bond]N = bis(3,5-dimethylpyrazol-1 yl)methane); R = Me, Et, or (i)Pr) are obtained when [Pd(N[bond]N)(C(6)F(5))Cl] is treated with KOH in the corresponding alcohol ROH and CO is bubbled through the solution. The reactions of [Pd(N[bond]N)(C(6)F(5))(OH)] (N[bond]N = bipy or Me(2)bipy) with CO(2), in tetrahydrofuran, lead to the formation of the binuclear carbonate complexes [(N[bond]N)(C(6)F(5))Pd(mu-eta(2) CO(3))Pd(C(6)F(5))(N[bond]N)]. Complexes [Pd(N[bond]N)(C(6)F(5))(OH)] react in alcohol with PhNCS to yield the corresponding N-phenyl-O-alkylthiocarbamate complexes [Pd(N[bond]N)(C(6)F(5))[SC(OR)NPh]]. Similarly, the reaction of [Pd(bipy)(C(6)F(5))(OH)] with PhNCO in methanol gives the N-phenyl-O methylcarbamate complex [Pd(bipy)(C(6)F(5))[NPhC(O)OR]]. The reactions of [(N[bond]N)Pd(C(6)F(5))(OH)] with PhNCS in the presence of Et(2)NH yield the corresponding thioureidometal complexes [Pd(N[bond]N)(C(6)F(5))[NPhCSNR(2)]]. The crystal structures of [Pd(tmeda)(C(6)F(5))(CO(2)Me)], [Pd(2)(Me(2)bipy)(2)(C(6)F(5))(2)(mu-eta(2)-CO(3))].2CH(2)Cl(2), and [Pd(tmeda)(C(6)F(5))[SC(OMe)NPh]] have been determined. PMID- 12767206 TI - 1H NMR studies of ligand and H/D exchange reactions of cis- and trans ([14]aneN4)(H2O)RhH2+ in aqueous solutions. AB - Substitution and exchange reactions of cis- and trans-L(1)(H(2)O)RhH(2+) (L(1) = 1,4,8,11-tetraazacyclotetradecane = [14]aneN(4)) were studied in aqueous solutions by UV-vis and (1)H NMR spectroscopies. At pH 1 and 25 degrees C, the substitution of SCN(-) for the coordinated molecule of water is rapid and thermodynamically favorable. Spectrophotometric determinations yielded the equilibrium constants K = 1.49 x 10(3) M(-1) (cis) and 1.44 x 10(3) (trans). (1)H NMR studies in D(2)O revealed a rapid dynamic process, interpreted as the exchange between coordinated water and X(-) (X = Cl, Br, or I). On the other hand, no line broadening was observed for the strongly bound ligands CN(-) and SCN(-). The complex trans-L(1)(D(2)O)RhH(2+) undergoes a base-catalyzed H/D exchange of the hydride in D(2)O with a rate constant of (1.45 +/- 0.02) x 10(3) M(-1) s(-1). The exchange in the cis isomer is very slow under similar conditions. The complex cis-[L(1)ClRhH](ClO(4)) crystallizes in the centrosymmetric Ponemacr; space group, unit cell dimensions a = 8.9805(11) A, b = 9.1598(11) A, c = 10.4081(13) A, alpha = 81.091(2) degrees, beta = 81.978(2) degrees, gamma = 88.850(2) degrees. The rhodium atom resides in a slightly distorted octahedral environment consisting of the four N atoms of the cyclam, a stereochemically active hydrogen, and a chlorine atom. PMID- 12767207 TI - Modified polyoxometalates: Hydrothermal syntheses and crystal structures of three novel reduced and capped Keggin derivatives decorated by transition metal complexes. AB - Three novel polyoxometalate derivatives decorated by transition metal complexes have been hydrothermally synthesized. Compound 1 consists of [PMo(VI)(6)Mo(V)(2)V(IV)(8)O(44)[Co (2,2'-bipy)(2)(H(2)O)](4)](3+) polyoxocations and [PMo(VI)(4-)Mo(V)(4)V(IV)(8)O(44)[Co(2,2'-bipy)(2)(H(2)O)](2)](3-) polyoxoanions, which are both built on mixed-metal tetracapped [PMo(8)V(8)O(44)] subunits covalently bonded to four or two [Co(2,2'-bpy)(2)(H(2)O)](2+) clusters via terminal oxo groups of the capping V atoms. Compound 2 is built on [PMo(VI)(8)V(IV)(6)O(42)[Cu(I)(phen)](2)](5-) clusters constructed from mixed metal bicapped [PMo(VI)(8)V(IV)(6)O(42)](7-) subunits covalently bonded to two [Cu(phen)](+) fragments in the similar way to 1. The structure of 3 is composed of [PMo(VI)(9)Mo(V)(3)O(40)](6-) units capped by two divalent Ni atoms via four bridging oxo groups. The crystal data for these are the following: C(120)H(126)Co(6)Mo(16)N(24)O(103)P(2)V(16) (1), triclinic P1, a = 15.6727(2) A, b = 17.3155(3) A, c = 19.5445(2) A, alpha = 86.1520(1) degrees, beta = 81.2010(1) degrees, gamma = 63.5970(1) degrees, Z = 1; C(120)H(85)Cu(6-)Mo(8)N(20)O(44)PV(6) (2), triclinic P1, a = 14.565(4) A, b = 15.899(3) A, c = 16.246(4) A, alpha = 116.289(2) degrees, beta = 103.084(2) degrees, gamma = 94.796(2) degrees, Z = 1; C(60)H(40)Mo(12)N(10)Ni(3)O(40)P (3), monoclinic P2(1)/c, a = 14.804(3) A, b = 22.137(4) A, c = 25.162(5) A, alpha = 90 degrees, beta = 98.59(3) degrees, gamma = 90 degrees, Z = 4. PMID- 12767208 TI - Organometallic[bond]polyoxometalate hybrid compounds: metallosalen compounds modified by Keggin type polyoxometalates. AB - Hybrid compounds with two functional centers consisting of a metallosalen moiety (M[bond]salen; M = Mn, Co, Ni, and Pd) connected by an alkylene bridging group to a lacunary Keggin type polyoxometalate were synthesized and characterized. In these metallosalen-polyoxometalate compounds (M[bond]salen[bond]POM) it was shown by the use of a combination of UV[bond]vis, (1)H NMR, EPR, XPS, and cyclic voltammetry measurements that the polyoxometalate exerts a significant intramolecular electronic effect on the metallosalen moiety leading to formation of an oxidized metallosalen moiety. For the Mn[bond]salen[bond]POM, the metallosalen center is best described as a metal[bond]salen cation radical species; that is, a localized "hole" is formed on the salen ligand. For the other M[bond]salen[bond]POM compounds, the metallosalen moiety can be described as a hybrid of a metal[bond]salen cation radical species and an oxidized metal[bond]salen species, that is, a delocalized "hole" is formed at the metallosalen center. It is proposed that these oxidized metallosalen centers are best characterized as stabilized charge transfer (metallosalen donor[bond]polyoxometalate acceptor) complexes despite the relatively large distance between the two functional centers. PMID- 12767209 TI - Complexation of uranium(VI) and samarium(III) with oxydiacetic acid: temperature effect and coordination modes. AB - The complexation of uranium(VI) and samarium(III) with oxydiacetate (ODA) in 1.05 mol kg(-1) NaClO(4) is studied at variable temperatures (25-70 degrees C). Three U(VI)/ODA complexes (UO(2)L, UO(2)L(2)(2-), and UO(2)HL(2)(-)) and three Sm(III)/ODA complexes (SmL(j)((3-2)(j)+) with j = 1, 2, 3) are identified in this temperature range. The formation constants and the molar enthalpies of complexation are determined by potentiometry and calorimetry. The complexation of uranium(VI) and samarium(III) with oxydiacetate becomes more endothermic at higher temperatures. However, the complexes become stronger due to increasingly more positive entropy of complexation at higher temperatures that exceeds the increase in the enthalpy of complexation. The values of the heat capacity of complexation (Delta C(p) degrees in J K(-1) mol(-1)) are 95 +/- 6, 297 +/- 14, and 162 +/- 19 for UO(2)L, UO(2)L(2)(2-), and UO(2)HL(2)(-), and 142 +/- 6, 198 +/- 14, and 157 +/- 19 for SmL(+), SmL(2)(-), and SmL(3)(3-), respectively. The thermodynamic parameters, in conjunction with the structural information from spectroscopy, help to identify the coordination modes in the uranium oxydiacetate complexes. The effect of temperature on the thermodynamics of the complexation is discussed in terms of the electrostatic model and the change in the solvent structure. PMID- 12767210 TI - Uranyl complexes with diamide ligands: a quantum mechanics study of chelating properties in the gas phase. AB - We report a quantum mechanical study on the complexes of UO(2)(2+) with diamide ligands L of malonamide and succinamide type, respectively, forming 6- and 7 chelate rings in their bidentate coordination to uranium. The main aims are to (i) assess how strong the chelate effect is (i.e., the preference for bi- versus monodentate binding modes of L), (ii) compare these ligands as a function of the chelate ring size, and (iii) assess the role of neutralizing counterions. For this purpose, we consider UO(2)L(2+), UO(2)L(2)(2+), UO(2)L(3)(2+), and UO(2)X(2)L type complexes with X(-) = Cl(-) versus NO(3)(-). Hartree-Fock and DFT calculations lead to similar trends and reveal the importance of saturation and steric repulsions ("strain") in the first coordination sphere. In the unsaturated UO(2)L(2+), UO(2)L(2)(2+), and UO(2)Cl(2)L complexes, the 7-ring chelate is preferred over the 6-ring chelate, and bidentate coordination is preferred over the monodentate one. However, in the saturated UO(2)(NO(3))(2)L complexes, the 6- and 7-chelating ligands have similar binding energies, and for a given ligand, the mono- and bidentate binding modes are quasi-isoenergetic. These conclusions are confirmed by the calculations of free energies of complexation in the gas phase. In condensed phases, the monodentate form of UO(2)X(2)L complexes should be further stabilized by coordination of additional ligands, as well as by interactions (e.g., hydrogen bonding) of the "free" carbonyl oxygen, leading to an enthalpic preference for this form, compared to the bidentate one. We also considered an isodesmic reaction exchanging one bidentate ligand L with two monoamide analogues, which reveals that the latter are clearly preferred (by 23 14 kcal/mol at the HF level and 24-12 kcal/mol at the DFT level). Thus, in the gas phase, the studied bidentate ligands are enthalpically disfavored, compared to bis-monodentate analogues. The contrast with trends observed in solution hints at the importance of "long range" forces (e.g., second shell interactions) and entropy effects on the chelate effect in condensed phases. PMID- 12767213 TI - Structure, topology, and dynamics of myristoylated recoverin bound to phospholipid bilayers. AB - Recoverin, a member of the EF-hand protein superfamily, serves as a calcium sensor in retinal rod cells. A myristoyl group covalently attached to the N terminus of recoverin facilitates its binding to retinal disk membranes by a mechanism known as the Ca(2+)-myristoyl switch. Samples of (15)N-labeled Ca(2+) bound myristoylated recoverin bind anisotropically to phospholipid membranes as judged by analysis of (15)N and (31)P chemical shifts observed in solid-state NMR spectra. On the basis of a (2)H NMR order parameter analysis performed on recoverin containing a fully deuterated myristoyl group, the N-terminal myristoyl group appears to be located within the lipid bilayer. Two-dimensional solid-state NMR ((1)H-(15)N PISEMA) spectra of uniformly and selectively (15)N-labeled recoverin show that the Ca(2+)-bound protein is positioned on the membrane surface such that its long molecular axis is oriented approximately 45 degrees with respect to the membrane normal. The N-terminal region of recoverin points toward the membrane surface, with close contacts formed by basic residues K5, K11, K22, K37, R43, and K84. This orientation of the membrane-bound protein allows an exposed hydrophobic crevice, near the membrane surface, to serve as a potential binding site for the target protein, rhodopsin kinase. Close agreement between experimental and calculated solid-state NMR spectra of recoverin suggests that membrane-bound recoverin retains the same overall three-dimensional structure that it has in solution. These results demonstrate that membrane binding by recoverin is achieved primarily by insertion of the myristoyl group inside the bilayer with apparently little rearrangement of the protein structure. PMID- 12767214 TI - The structure of a functional unit from the wall of a gastropod hemocyanin offers a possible mechanism for cooperativity. AB - Structure-function relationships in a molluscan hemocyanin have been investigated by determining the crystal structure of the Rapana thomasiana (gastropod) hemocyanin functional unit RtH2e in deoxygenated form at 3.38 A resolution. This is the first X-ray structure of an unit from the wall of the molluscan hemocyanin cylinder. The crystal structure of RtH2e demonstrates molecular self-assembly of six identical molecules forming a regular hexameric cylinder. This suggests how the functional units are ordered in the wall of the native molluscan hemocyanins. The molecular arrangement is stabilized by specific protomer-to-protomer interactions, which are probably typical for the functional units building the wall of the cylinders. A molecular mechanism for cooperative dioxygen binding in molluscan hemocyanins is proposed on the basis of the molecular interactions between the protomers. In particular, the deoxygenated RtH2e structure reveals a tunnel leading from two opposite sides of the molecule to the active site. The tunnel represents a possible entrance pathway for dioxygen molecules. No such tunnels have been observed in the crystal structure of the oxy-Odg, a functional unit from the Octopus dofleini (cephalopod) hemocyanin in oxygenated form. PMID- 12767215 TI - An improved solution structure for psi-conotoxin PiiiE. AB - A revised, high-resolution structure of psi-conotoxin Piiie (psi-Piiie), a noncompetitive inhibitor of the nicotinic acetylcholine receptor (nAChR), produced through the use of NMR and molecular modeling calculations is presented. The original structures of psi-Piiie had a relatively high degree of disorder, particularly in the conformation of the disulfide bridges. Our studies utilized (13)C-labeling of selected cysteine residues allowing the resolution of all problems of resonance overlap for the cysteine residues. The improved data were used to produce a new set of structures by a molecular modeling process incorporating relaxation matrix methods for the determination of interproton distance restraints and a combination of distance geometry and simulated annealing for structure generation. The structures produced are very well converged with the RMSD of backbone atom positions of the main body of the peptide improving from 0.73 to 0.13 A. Other indicators of correlation with the experimental data and quality of covalent geometry showed significant improvement in the new structures. The overall conformation of the peptide backbone is similar between the two determinations with the exception of the N-terminus. This difference leads to a significant effect on the predicted distribution of positive charge within psi-Piiie, a property likely to influence interpretation of future mutational studies. PMID- 12767216 TI - Characterization and three-dimensional structure determination of psi-conotoxin Piiif, a novel noncompetitive antagonist of nicotinic acetylcholine receptors. AB - A novel inhibitor of nicotinic acetylcholine receptors (nAChRs), psi-conotoxin Piiif, was isolated from the venom of Conus purpurascens and found to have the sequence GOOCCLYGSCROFOGCYNALCCRK-NH2. The sequence is highly homologous to that of psi-conotoxin Piiie, a previously identified noncompetitive inhibitor of Torpedo electroplax nAChR, also isolated from C. purpurascens. Both psi conotoxins block Torpedo and mouse nicotinic acetylcholine receptors (nAChRs), but psi-Piiif is less potent by a factor of 10(1)-10(2). A high-resolution structure of psi-Piiif was determined by NMR and molecular modeling calculations. Psi-Piiif analogues containing [(13)C]-labeled cysteine at selected positions were synthesized to resolve spectral overlap of Cys side chain proton signals. The structures are well-converged, with backbone atom position RMSDs of 0.21 A for the main body of the peptide between residues 4 and 22 and 0.47 A for all residues. The overall backbone conformation is closely similar to psi-Piiie, the main difference being in the degree of conformational disorder at the two termini. Psi-Piiie and psi-Piiif have similar locations of positive charge density, although psi-Piiif has a lower overall charge. One disulfide bridge of psi-Piiif appears to undergo dynamic conformational fluctuations based on both the model and on experimental observation. Chimeras in which the three intercysteine loops were swapped between psi-Piiie and psi-Piiif were tested for inhibitory activity against Torpedo nAChRs. The third loop, which contains no charged residues in either peptide, is the prime determinant of potency in these psi-conotoxins. PMID- 12767217 TI - Sulfaphenazole derivatives as tools for comparing cytochrome P450 2C5 and human cytochromes P450 2Cs: identification of a new high affinity substrate common to those enzymes. AB - The inhibitory effects of a series of sulfaphenazole (SPA) derivatives were studied on two modified forms of rabbit liver cytochrome P450 2C5 (CYP2C5), CYP2C5dH, and structurally characterized CYP2C5/3LVdH and compared to the previously described effects of these compounds on human CYP2C8, 2C9, 2C18, and 2C19. SPA and other negatively charged compounds that potently inhibit CYP2C9 had very little effect on CYP2C5dH, whereas neutral, N-alkylated derivatives exhibited IC50 values between 8 and 22 microM. One of the studied compounds, 4, that derives from SPA by replacement of its NH(2) substituent with a methyl group and by N-methylation of its sulfonamide moiety, acted as a good substrate for all CYP2Cs used in this study. Hydroxylation of the benzylic methyl of 4 is the major reaction catalyzed by all of these CYP2C proteins, whereas hydroxylation of the N phenyl group of 4 was observed as a minor reaction. CYP2C5dH, 2C5/3LVdH, 2C9, 2C18, and 2C19 are efficient catalysts for the benzylic hydroxylation of 4, with K(m) values between 5 and 13 microM and k(cat) values between 16 and 90 min(-1). The regioselectivity observed for oxidation of 4 by CYP2C5/3LVdH was easily interpreted on the basis of the existence of two different binding modes of 4 characterized in the experimentally determined structure of the complexes of CYP2C5/3LVdH with 4 described in the following paper [Wester, M. R. et al. (2003) Biochemistry 42, 6370-6379]. PMID- 12767218 TI - Structure of a substrate complex of mammalian cytochrome P450 2C5 at 2.3 A resolution: evidence for multiple substrate binding modes. AB - The structure of rabbit microsomal cytochrome P450 2C5/3LVdH complexed with a substrate, 4-methyl-N-methyl-N-(2-phenyl-2H-pyrazol-3-yl)benzenesulfonamide (DMZ), was determined by X-ray crystallography to 2.3 A resolution. Substrate docking studies and electron density maps indicate that DMZ binds to the enzyme in two antiparallel orientations of the long axis of the substrate. One orientation places the principal site of hydroxylation, the 4-methyl group, 4.4 A from the heme Fe, whereas the alternate conformation positions the second, infrequent site of hydroxylation at >5.9 A from the heme Fe. Comparison of this structure to that obtained previously for the enzyme indicates that the protein closes around the substrate and prevents open access of water from bulk solvent to the heme Fe. This reflects a approximately 1.5 A movement of the F and G helices relative to helix I. The present structure provides a complete model for the protein from residues 27-488 and defines two new helices F' and G'. The G' helix is likely to contribute to interactions of the enzyme with membranes. The relatively large active site, as compared to the volume occupied by the substrate, and the flexibility of the enzyme are likely to underlie the capacity of drug-metabolizing enzymes to metabolize structurally diverse substrates of different sizes. PMID- 12767219 TI - Two conformational states of Turkey ovomucoid third domain at low pH: three dimensional structures, internal dynamics, and interconversion kinetics and thermodynamics. AB - Turkey ovomucoid third domain (OMTKY3) is shown to exist at low pH as two distinctly folded, interconverting conformations. Activation parameters were determined for the transition, and these were of the type reported previously for cis/trans isomerizations of prolyl peptide bonds. Multidimensional, multinuclear NMR spectroscopy was used to determine the three-dimensional structure of each of the two states of P(5)-Pro(14)Asp OMTKY3 at pH 2.5 and 25 degrees C, under conditions where the two states have equal populations with interchange rates of 0.25 s(-1). The results showed that the two states differ by cis/trans isomerization of the P(8)-Tyr(11)-P(7)-Pro(12) peptide bond, which is cis in the conformer dominant at neutral pH and trans in the conformer appearing at low pH. The major structural differences were found to be in the region of the reactive site loop. The core of the protein, including the antiparallel beta-sheet and a alpha-helix, is preserved in both structures. The state with the cis peptide bond is similar to previously reported structures of OMTKY3 determined by NMR spectroscopy and X-ray crystallography. The cis-to-trans transition results in the relocation of the aromatic ring of P(8)-Tyr(11), disrupts many interactions between the alpha-helix and the reactive-site loop, and leads to more open spacing between this loop and the alpha-helix. In addition, the configurations of two of the three disulfide bonds, P(11)-Cys(8)- P(20)'-Cys(38), and P(3)-Cys(16)- P(17)'-Cys(35), are altered such that the C(alpha)-C(alpha) distances for each disulfide bridge are longer by approximately 1 A in the trans state than in the cis. Mutations at P(1)-Leu(18), P(6)-Lys(13), and P(5)-Pro(14) influence the position of the cis <= => trans equilibrium. In P(1)-Leu(18)Xxx OMTKY3 mutants, the trans state is more favored by P(1)-Gly(18) than by Ala(18) or Leu(18); in P(6)-Lys(13)Xxx OMTKY3 mutants, the trans state is more favored by P(6)-Glu(13) and P(6)-Asp(13) than Lys(13) or His(13). Stabilization of the trans state in P(5)-Pro(14)Xxx OMTKY3 mutants follows the series Xxx = Gly > Asp > Glu > Ala approximately equal His > Pro. In comparing the state with the trans peptide bond to that with the cis, the pK(a) values of P(12)-Asp(7) and P(1)'-Glu(19) are higher and those of P(9)-Glu(10) and P(25)'-Glu(43) are lower. The pK(a) values of other titrating groups in the molecule are similar in both conformational states. These pK(a) changes underlie the pH dependence of the conformational equilibrium and can be explained in part by observed structural differences. (15)N transverse relaxation results indicate that residues P(6)-Lys(13)-P(3) Cys(16) in the trans state undergo a dynamic process on the microsecond millisecond time scale not present in the cis state. PMID- 12767220 TI - Crystal structure of the human GGA1 GAT domain. AB - GGAs are a family of vesicle-coating regulatory proteins that function in intracellular protein transport. A GGA molecule contains four domains, each mediating interaction with other proteins in carrying out intracellular transport. The GAT domain of GGAs has been identified as the structural entity that binds membrane-bound ARF, a molecular switch regulating vesicle-coat assembly. It also directly interacts with rabaptin5, an essential component of endosome fusion. A 2.8 A resolution crystal structure of the human GGA1 GAT domain is reported here. The GAT domain contains four helices and has an elongated shape with the longest dimension exceeding 80 A. Its longest helix is involved in two structural motifs: an N-terminal helix-loop-helix motif and a C terminal three-helix bundle. The N-terminal motif harbors the most conservative amino acid sequence in the GGA GAT domains. Within this conserved region, a cluster of residues previously implicated in ARF binding forms a hydrophobic surface patch, which is likely to be the ARF-binding site. In addition, a structure-based mutagenesis-biochemical analysis demonstrates that the C-terminal three-helix bundle of this GAT domain is responsible for the rabaptin5 binding. These structural characteristics are consistent with a model supporting multiple functional roles for the GAT domain. PMID- 12767221 TI - Position-dependence of stabilizing polar interactions of asparagine in transmembrane helical bundles. AB - Recent studies with model peptides and statistical analyses of the crystal structures of membrane proteins have shown that buried polar interactions contribute significantly to the stabilization of the three-dimensional structures of membrane proteins. Here, we probe how the location of these polar groups along the transmembrane helices affect their free energies of interaction. Asn residues were placed singly and in pairs at three positions within a model transmembrane helix, which had previously been shown to support the formation of trimers in micelles. The model helix was designed to form a transmembrane coiled coil, with Val side chains at the "a" positions of the heptad repeat. Variants of this peptide were prepared in which an Asn residue was introduced at one or more of the "a" positions, and their free energies of association were determined by analytical ultracentrifugation. When placed near the middle of the transmembrane helix, the formation of trimers was stabilized by at least -2.0 kcal/mol per Asn side chain. When the Asn was placed at the interface between the hydrophobic and polar regions of the peptide, the substitution was neither stabilizing nor destabilizing (0.0 +/- 0.5 kcal/mol of monomer). Finally, it has previously been shown that a Val-for-Asn mutation in a water-soluble coiled coil destabilizes the structure by approximately 1.5 kcal/mol of monomer [Acharya, A., et al. (2002) Biochemistry 41, 14122-14131]. Thus, the headgroup region of a micelle appears to have a conformational impact intermediate between that of bulk water and the apolar region of micelle. A similarly large dependence on the location of the polar residues was found in a statistical survey of helical transmembrane proteins. The tendency of different types of residues to be buried in the interiors versus being exposed to lipids was analyzed. Asn and Gln show a very strong tendency to be buried when they are located near the middle of a transmembrane helix. However, when placed near the ends of transmembrane helices, they show little preference for the surface versus the interior of the protein. These data show that Asn side chains within the apolar region of the transmembrane helix provide a significantly larger driving force for association than Asn residues near the apolar/polar interface. Thus, although polar interactions are able to strongly stabilize the folding of membrane proteins, the energetics of association depend on their location within the hydrophobic region of a transmembrane helix. PMID- 12767223 TI - Effect of temperature and the F27W mutation on the Ca2+ activated structural transition of trout cardiac troponin C. AB - The Ca(2+) sensitivity of cardiac contractile element is reduced at lower temperatures, in contrast to that in fast skeletal muscle. Cardiac troponin C (cTnC) replacement in mammalian skinned fibers showed that TnC plays a critical role in this phenomenon (Harrison and Bers, (1990), Am. J. Physiol. 258, C282-8). Understanding the differences in affinity and structure between cTnCs from cold adapted ectothermic species and mammals may bring new insights into how the different isoforms provide different resistances to cold. We followed the Ca(2+) titration to the regulatory domain of rainbow trout cTnC by NMR (wild type at 7 and 30 degrees C and F27W mutant at 30 degrees C) and fluorescence (F27W mutant, at 7 and 30 degrees C) spectroscopies. Using NMR spectroscopy, we detected Ca(2+) binding to site I of trout cTnC at high concentrations. This places trout cTnC between mammalian cTnC, in which site I is completely inactive, and skeletal TnC, in which site I binds Ca(2+) during muscle activation, and which is not as much affected by lower temperatures. This binding was seen both at 7 and at 30 degrees C. Despite the low Ca(2+) affinity, trout TnC site I may increase the likelihood of an opening of the regulatory domain, thus increasing the affinity for TnI. This way, it may be responsible for trout cTnC's capacity to function at lower temperatures. PMID- 12767222 TI - Role of myristylation in HIV-1 Gag assembly. AB - Assembly of the human immunodeficiency virus type 1 (HIV-1) first occurs on the plasma membrane of host cells where binding is driven by strong electrostatic interactions between the N-terminal matrix (MA) domain of the structural precursor polyprotein, Gag, and the membrane. MA is also myristylated, but the exact role this modification plays is not clear. In this study, we compared the protein oligomerization and membrane binding properties of Myr(+) and Myr(-) Gag(MA) expressed in COS-1 cells. Sedimentation studies in solution showed that both the myristylated Gag precursor and the mature MA product were detected in larger complexes than their unmyristylated counterparts, and the myristylated MA protein bound liposomes with approximately 3-fold greater affinity than unmyristylated MA. Aromatic residues near the N-terminal region of the MA protein were more accessible to chymotrypsin in the unmyristylated form and, consistent with this, an epitope in the N-terminal region was more exposed. Moreover, the cyclophilin binding site in the CA domain downstream of MA was more accessible in the unmyristylated Gag protein, while the Tsg101 binding site in the C-terminal region was equally available in the unmyristylated and myristylated Gag proteins. Taken together, our results suggest that myristylation promotes assembly by inducing conformational changes and facilitating MA multimerization. This observation offers a novel role for myristylation. PMID- 12767224 TI - A triple mutant of the Drosophila ERR confers ligand-induced suppression of activity. AB - The steroid hormone (NR3) subfamily of nuclear receptors was until recently believed to be restricted to deuterostomes. However, a novel nuclear receptor belonging to the NR3 subfamily was recently identified in the Drosophila melanogaster genome, indicating the existence of an ancestor before the evolutionary split of deuterostomes and protostomes. This receptor, termed the Drosophila estrogen-related receptor (dERR), most closely resembles the human and mouse estrogen-related receptors (ERRs) in both the DNA binding domain (DBD) (approximately 85% identical) and the ligand binding domain (LBD) (approximately 35% identical). Here we describe the functional analysis and rational design of ligand responsive dERR mutants created by protein engineering of the LBD. On the basis of homology modeling, three amino acid residues in the LBD were identified and mutated to enable ligand-dependent suppression of transcriptional activity. Our results show that the Y295A/T333I/Y365L triple mutant is significantly suppressed by the known ERR inverse agonists 4-hydroxytamoxifen (OHT) and diethylstilbestrol (DES), in comparison to the wild-type dERR receptor, which was inefficiently suppressed by these substances. The coactivator mGRIP-1 (mouse glucocorticoid receptor interacting protein 1) was shown to significantly increase the activity of the triple mutant in transfection experiments, and the addition of OHT resulted in an efficient suppression of the activity. Accordingly, the ability to functionally interact with a coactivator is still maintained by the Y295A/T333I/Y365L mutant. These findings demonstrate the potential of using rational design and engineering of the LBD to study the function of a nuclear receptor lacking identified ligands. PMID- 12767225 TI - Characterization of two independent amino acid substitutions that disrupt the DNA repair functions of the yeast Apn1. AB - The members of the Endo IV family of DNA repair enzymes, including Saccharomyces cerevisiae Apn1 and Escherichia coli endonuclease IV, possess the capacity to cleave abasic sites and to remove 3'-blocking groups at single-strand breaks via apurinic/apyrimidinic (AP) endonuclease and 3'-diesterase activities, respectively. In addition, Endo IV family members are able to recognize and incise oxidative base damages on the 5'-side of such lesions. We previously identified eight amino acid substitutions that prevent E. coli endonuclease IV from repairing damaged DNA in vivo. Two of these substitutions were glycine replacements of Glu145 and Asp179. Both Glu145 and Asp179 are among nine amino acid residues within the active site pocket of endonuclease IV that coordinate the position of a trinuclear Zn cluster required for efficient phosphodiester bond cleavage. We now report the first structure-function analysis of the eukaryotic counterpart of endonuclease IV, yeast Apn1. We show that glycine substitutions at the corresponding conserved amino acid residues of yeast Apn1, i.e., Glu158 and Asp192, abolish the biological function of this enzyme. However, these Apn1 variants do not exhibit the same characteristics as the corresponding E. coli mutants. Indeed, the Apn1 Glu158Gly mutant, but not the E. coli endonuclease IV Glu145Gly mutant, is able to bind DNA. Moreover, Apn1 Asp192Gly completely lacks enzymatic activity, while the activity of the E. coli counterpart Asp179Gly is reduced by approximately 40-fold. The data suggest that although yeast Apn1 and E. coli endonuclease IV exhibit a high degree of structural and functional similarity, differences exist within the active site pockets of these two enzymes. PMID- 12767226 TI - Eight amino acids form the ATP recognition site of Na(+)/K(+)-ATPase. AB - Point mutations of a part of the H(4)-H(5) loop (Leu(354)-Ile(604)) of Na(+)/K(+) ATPase have been used to study the ATP and TNP-ATP binding affinities. Besides the previously reported amino acid residues Lys(480), Lys(501), Gly(502), and Cys(549), we have found four more amino acid residues, viz., Glu(446), Phe(475), Gln(482), and Phe(548), completing the ATP-binding pocket of Na(+)/K(+)-ATPase. Moreover, mutation of Arg(423) has also resulted in a large decrease in the extent of ATP binding. This residue, localized outside the binding pocket, seems to play a key role in supporting the proper structure and shape of the binding site, probably due to formation of a hydrogen bond with Glu(472). On the other hand, only some minor effects were caused by mutations of Ile(417), Asn(422), Ser(445), and Glu(505). PMID- 12767227 TI - Identification of substrate contact residues important for the allosteric regulation of phosphofructokinase from Eschericia coli. AB - The side chains of Escherichia coli phosphofructokinase (EcPFK) that interact with bound substrate, fructose 6-phosphate (Fru-6-P), are examined for their potential roles in allosteric regulation. Mutations that severely decrease Fru-6 P affinity and/or k(cat)/K(m) were created at each contact residue, with the exception of the catalytic base, D127. Even though Fru-6-P affinity was greatly decreased for R162E, M169A, E222A/H223A, and R243E, the mutated proteins retained the ability to be activated by MgADP and inhibited by phosphoenolpyruvate (PEP). R252E did not show an allosteric response to either MgADP or PEP. The H249E mutation retained MgADP activation but did not respond to PEP. R72E, T125A, and R171E maintained allosteric inhibition by PEP. Both R72E and T125A displayed a MgADP-dependent decrease in k(cat) but no MgADP-dependent K-type effects. R171E maintained MgADP-dependent K-type activation but also displayed a MgADP-dependent decrease in k(cat). Localization of mutations that alter MgADP activation near the transferred phosphate group indicates the importance of the 1-methoxy region of Fru-6-P in allosteric regulation by MgADP. A region near the 6'-phosphate may be similarly important for PEP inhibition. R252 is uniquely positioned between the 1'- and 6'-phosphates of bound Fru-1,6-BP, and the mutation at this position may alter both allosterically responsive regions. The differential functions of specific regions in the Fru-6-P contact residues support different mechanisms for allosteric activation and inhibition. In addition, the lack of correlation between mutations that decrease Fru-6-P affinity and those that abolish allosteric communications supports the independence of affinity and allosteric coupling. PMID- 12767228 TI - Testing of the additivity-based protein sequence to reactivity algorithm. AB - The standard free energies of association (or equilibrium constants) are predicted for 11 multiple variants of the turkey ovomucoid third domain, a member of the Kazal family of protein inhibitors, each interacting with six selected enzymes. The equilibrium constants for 38 of 66 possible interactions are strong enough to measure, and for these, the predicted and measured free energies are compared, thus providing an additional test of the additivity-based sequence to reactivity algorithm. The test appears to be unbiased as the 11 variants were designed a decade ago to study furin inhibition and the specificity of furin differs greatly from the specificities of our six target enzymes. As the contact regions of these inhibitors are highly positive, nonadditivity was expected. Of the 11 variants, one does not satisfy the restriction that either P(2) Thr or P(1)' Glu should be present and all three measurable results on it, as expected, are nonadditive. For the remaining 35 measurements, 22 are additive, 12 are partially additive, and only one is (slightly) nonadditive. These results are comparable to those obtained for a set of 398 equilibrium constants for natural variants of ovomucoid third domains. The expectation that clustering of charges would be nonadditive is modified to the expectation that major nonadditivity will be observed only if the combining sites in both associating proteins involve large charge clusters of the opposite sign. It is also shown here that an analysis of a small variant set can be accomplished with a smaller subset, in this case 13 variants, rather than the whole set of 191 members used for the complete algorithm. PMID- 12767230 TI - Discovery and characterization of cooperative ligand binding in the adaptive region of interleukin-2. AB - The cytokine hormone interleukin-2 (IL-2) contains a highly adaptive region that binds small, druglike molecules. The binding properties of this adaptive region have been explored using a "tethering" method that relies on the formation of a disulfide bond between the protein and small-molecule ligands. Using tethering, surface plasmon resonance (SPR), and X-ray crystallography, we have discovered that the IL-2 adaptive region contains at least two cooperative binding sites where the binding of a first ligand to one site promotes or antagonizes the binding of a second ligand to the second site. Cooperative energies of interaction of -2 kcal/mol are observed. The observation that the adaptive region contains two adjacent sites may lead to the development of tight-binding antagonists of a protein-protein interaction. Cooperative ligand binding in the adaptive region of IL-2 underscores the importance of protein dynamics in molecular recognition. The tethering approach provides a novel and general strategy for discovering such cooperative binding interactions in specific, flexible regions of protein structure. PMID- 12767229 TI - Ligand specificity in the CRAL-TRIO protein family. AB - Intracellular trafficking of hydrophobic ligands is often mediated by specific binding proteins. The CRAL-TRIO motif is common to several lipid binding proteins including the cellular retinaldehyde binding protein (CRALBP), the alpha tocopherol transfer protein (alpha-TTP), yeast phosphatidylinositol transfer protein (Sec14p), and supernatant protein factor (SPF). To examine the ligand specificity of these proteins, we measured their affinity toward a variety of hydrophobic ligands using a competitive [(3)H]-RRR-alpha-tocopherol binding assay. Alpha-TTP preferentially bound RRR-alpha-tocopherol over all other tocols assayed, exhibiting a K(d) of 25 nM. Binding affinities of other tocols for alphaTTP closely paralleled their ability to inhibit in vitro intermembrane transfer and their potency in biological assays. All other homologous proteins studied bound alpha-tocopherol but with pronouncedly weaker (> 10-fold) affinities than alpha-TTP. Sec14p demonstrated a K(d) of 373 nM for alpha tocopherol, similar to that for its native ligand, phosphatidylinositol (381 nM). Human SPF had the highest affinity for phosphatidylinositol (216 nM) and gamma tocopherol (268 nM) and significantly weaker affinity for alpha-tocopherol (K(d) 615 nM). SPF bound [(3)H]-squalene more weakly (879 nM) than the other ligands. Our data suggest that of all known CRAL-TRIO proteins, only alphaTTP is likely to serve as the physiological mediator of alpha-tocopherol's biological activity. Further, ligand promiscuity observed within this family suggests that caution should be exercised when suggesting protein function(s) from measurements utilizing a single ligand. PMID- 12767231 TI - The role of the protein core in the inhibitory power of the classic serine protease inhibitor, chymotrypsin inhibitor 2. AB - A synthetic cyclic peptide, reported to be a tight-binding inhibitor of serine proteases, is instead found to be a good substrate, as is the linear peptide of the same sequence. Both of the peptides, designed to mimic the binding loop of chymotrypsin inhibitor 2 (CI2), were cleaved by subtilisin primarily at the CI2 reactive-site Met-59-Glu-60 bond, revealing that the sequence, in the absence of the structural context of the inhibitor, provides sufficient specificity for hydrolysis of this bond. Insights from the crystal structure of the CI2/subtilisin complex, together with biochemical analysis of a CI2 Gly-83 deletion mutant, have allowed us to identify key features that make CI2 an effective inhibitor, while the cyclic and linear peptides are substrates. PMID- 12767232 TI - Architecture of the Qo site of the cytochrome bc1 complex probed by superoxide production. AB - Although several X-ray structures have been determined for the mitochondrial cytochrome (cyt) bc(1) complex, none yet shows the position of the substrate, ubiquinol, in the quinol oxidase (Q(o)) site. In this study, the interaction of molecular oxygen with the reactive intermediate Q(o) semiquinone is used to probe the Q(o) site. It has been known for some time that partial turnover of the cyt bc(1) complex in the presence of antimycin A, a Q(i) site inhibitor, results in accumulation of a semiquinone at the Q(o) site, which can reduce O(2) to superoxide (O(2)(*)(-)). It was more recently shown that myxothiazol, which binds close to the cyt b(L) heme in the proximal Q(o) niche, also induces O(2)(*)(-) production. In this work, it is shown that, in addition to myxothiazol, a number of other proximal Q(o) inhibitors [including (E)-beta-methoxyacrylate-stilbene, mucidin, and famoxadone] also induce O(2)(*)(-) production in the isolated yeast cyt bc(1) complex, at approximately 50% of the V(max) observed in the presence of antimycin A. It is proposed that proximal Q(o) site inhibitors induce O(2)(*)(-) production because they allow formation, but not oxidation, of the semiquinone at the distal niche of the Q(o) site pocket. The apparent K(m) for ubiquinol at the Q(o) site in the presence of proximal Q(o) site inhibitors suggests that the "distal niche" of the Q(o) pocket can act as a fully independent quinol binding and oxidation site. Together with the X-ray structures, these results suggest substrate ubiquinol binds in a fashion similar to that of stigmatellin with H bonds between H161 of the Rieske iron-sulfur protein and E272 of the cyt b protein. When modeled in this way, mucidin and ubiquinol can bind simultaneously to the Q(o) site with virtually no steric hindrance, whereas progressively bulkier inhibitors exhibit increasing overlap. The fact that partial turnover of the Q(o) site is possible even with bound proximal Q(o) site inhibitors is consistent with the participation of two separate functional Q(o) binding niches, occupied simultaneously or sequentially. PMID- 12767233 TI - Role of the interdomain linker probed by kinetics of CO ligation to an endothelial nitric oxide synthase mutant lacking the calmodulin binding peptide (residues 503-517 in bovine). AB - Oxygenase and reductase domains in nitric oxide synthase are linked by a peptide region that binds calmodulin. Here we study the effects of modifying the length of the interdomain linker in a deletion mutant lacking 15 amino acids (residues 503-517) in bovine eNOS. The kinetics of CO ligation with the mutant were determined in the presence and absence of tetrahydrobiopterin and arginine and compared with the CO binding kinetics of wild-type eNOS and the eNOS oxygenase domain. In the mutant, electron flow is interrupted. The association kinetics of CO with both mutant and wild-type eNOS can be approximated with two kinetic phases, but the relative proportions change in the mutant. Both the abrogation of electron flow in the mutant and the differences in CO binding may be explained by an alteration in the docking of the FMN domain to the heme domain. We propose that the calmodulin binding residues form a helix that is critical for the proper alignment of the adjacent reductase and oxygenase domains within the active eNOS dimer in achieving proper electron transfer between them. PMID- 12767234 TI - Stability of intramolecular DNA quadruplexes: comparison with DNA duplexes. AB - We have determined the stability of intramolecular quadruplexes that are formed by a variety of G-rich sequences, using oligonucleotides containing appropriately placed fluorophores and quenchers. The stability of these quadruplexes is compared with that of the DNA duplexes that are formed on addition of complementary C-rich oligonucleotides. We find that the linkers joining the G tracts are not essential for folding and can be replaced with nonnucleosidic moieties, though their sequence composition profoundly affects quadruplex stability. Although the human telomere repeat sequence d[G(3)(TTAG(3))(3)] folds into a quadruplex structure, this forms a duplex in the presence of the complementary C-rich strand at physiological conditions. The Tetrahymena sequence d[G(4)(T(2)G(4))(3)], the sequence d[G(3)(T(2)G(3))(3)], and sequences related to regions of the c-myc promoter d(G(4)AG(4)T)(2) and d(G(4)AG(3)T)(2) preferentially adopt the quadruplex form in potassium-containing buffers, even in the presence of a 50-fold excess of their complementary C-rich strands, though the duplex predominates in the presence of sodium. The HIV integrase inhibitor d[G(3)(TG(3))(3)] forms an extremely stable quadruplex which is not affected by addition of a 50-fold excess of the complementary C-rich strand in both potassium and sodium-containing buffers. Replacing the TTA loops of the human telomeric repeat with AAA causes a large decrease in quadruplex stability, though a sequence with AAA in the first loop and TTT in the second and third loops is slightly more stable. PMID- 12767235 TI - Novel processing of beta-amyloid precursor protein catalyzed by membrane type 1 matrix metalloproteinase releases a fragment lacking the inhibitor domain against gelatinase A. AB - In various mammalian cell lines, beta-amyloid precursor protein (APP) is proteolytically processed to release its NH(2)-terminal extracellular domain as a soluble APP (sAPP) that contains the inhibitor domain against gelatinase A. To investigate roles of sAPP in the regulation of gelatinase A activity, we examined the correlation between the activation of progelatinase A and processing of APP. We found that stimulation of HT1080 fibrosarcoma cells with concanavalin A led to an activation of endogenous progelatinase A and to a novel processing of APP, which releases a COOH-terminally truncated form of sAPP (sAPPtrc) into the culture medium. Reverse zymographic analysis showed that sAPPtrc lacked an inhibitory activity against gelatinase A. Analyses of production of sAPPtrc in the presence of various metalloproteinase inhibitors showed that membrane type 1 matrix metalloproteinase (MT1-MMP), an activator of progelatinase A, is most likely responsible for the production of sAPPtrc. When the concanavalin A stimulated HT1080 cells were cultured in the condition that inhibited MT1-MMP activity, sAPP and APP were associated with the extracellular matrix deposited by the cells, whereas these gelatinase A inhibitors in the matrix were displaced by sAPPtrc after exertion of MT1-MMP activity. Taken together, these data support a model in which MT1-MMP-catalyzed release of sAPPtrc leads to reduction of the extracellular matrix-associated gelatinase A inhibitor, sAPP, thus making it feasible for gelatinase A to exert proteolytic activity only near its activator, MT1-MMP. PMID- 12767236 TI - Ligand binding dynamics to the heme domain of the oxygen sensor Dos from Escherichia coli. AB - In the heme-based oxygen sensor Dos from Escherichia coli, one of the axial ligands (Met 95) of a six-coordinate heme can be replaced by external ligands such as O(2), NO, and CO, which causes a switch in phosphodiesterase activity. To gain insight into the bidirectional switching mechanism, we have studied the interaction of ligands with the sensor domain DosH by flash photolysis experiments with femtosecond time resolution. The internal ligand can be photodissociated from the ferrous heme and recombines with time constants of 7 and 35 ps. This is somewhat slower than recombination of the external ligands NO, with which picosecond rebinding occurs with unprecedented efficiency (>99%) with a predominant phase of approximately 5 ps, and O(2) (97% in 5 ps, Liebl, U., Bouzhir-Sima, L., Negrerie, M., Martin, J.-L., and Vos, M. H. (2002) Proc. Natl. Acad. Sci. U.S.A. 99, 12771-12776). Dissociated CO displays geminate rebinding in 1.5 ns with a very high yield (60%). Together these results indicate that the heme environment provides a very tight pocket for external ligands, presumably preventing frequent switching events. Additional CO dissociation and rebinding experiments on a longer time scale reveal that (a) Met 95 binding, in 100 micros, occurs in competition with bimolecular CO binding, and (b) subsequent replacement of Met 95 by CO on the millisecond time scale occurs faster than in rapid-mixing experiments, suggesting a slow further relaxation. A minimal ligand binding model is proposed that suggests that Met 95 displacement from the heme is facilitated by the presence of an external ligand in the heme environment. Furthermore, the orders of magnitude difference between Met 95 binding after dissociation of internal and external ligands, as well as the spectral characteristics of photodissociation intermediates, indicate substantial rearrangement of the heme environment associated with ligand sensing. Further remarkable observations include evidence for stable (>4 ns) photooxidation of six-coordinate ferrous heme, with a quantum yield of 4-8%. PMID- 12767237 TI - Autophosphorylation of threonine 485 in the activation loop is essential for attaining eIF2alpha kinase activity of HRI. AB - In heme deficiency, protein synthesis is inhibited by the activation of the heme regulated eIF2alpha kinase (HRI) through its multiple autophosphorylation. Autophosphorylation sites in HRI were identified in order to investigate their functions. We found that there were eight major tryptic phosphopeptides of HRI activated in heme deficiency. In this report we focused on the role of autophosphorylation at Thr483 and Thr485 in the activation loop of HRI. Disruption of the autophosphorylation of Thr485, but not Thr483, resulted in a lower autokinase activity and locked Thr485Ala HRI in a hypophosphorylated state. Most importantly, autophosphorylation of Thr485, but not Thr483, was essential for attaining eIF2alpha kinase activity of HRI. In addition, autophosphorylation of Thr485 was necessary for arsenite-induced activation of the eIF2alpha kinase activity of HRI, while autophosphorylation at Thr483 was not required for activation by arsenite. The function of Thr490, another conserved Thr residue in the activation loop of HRI, was also investigated. Mutations of Thr490 to either Ala or Asp resulted in reduced autokinase activity and loss of eIF2alpha kinase activity in heme deficiency or upon arsenite treatment. Since Thr490 was not identified as an autophosphorylated site, it is likely that Thr490 itself might be critical for the catalytic activity of HRI. Importantly, Thr485 was very poorly phosphorylated in Thr490 mutant HRI. Collectively, our results demonstrate that autophosphorylation of Thr485 is essential for the hyperphosphorylation and activation of HRI and is required for the acquisition of the eIF2alpha kinase activity. PMID- 12767238 TI - Mechanism of lipid bilayer disruption by the human antimicrobial peptide, LL-37. AB - LL-37 is an amphipathic, alpha-helical, antimicrobial peptide. (15)N chemical shift and (15)N dipolar-shift spectroscopy of site-specifically labeled LL-37 in oriented lipid bilayers indicate that the amphipathic helix is oriented parallel to the surface of the bilayer. This surface orientation is maintained in both anionic and zwitterionic bilayers and at different temperatures and peptide concentrations, ruling out a barrel-stave mechanism for bilayer disruption by LL 37. In contrast, electrostatic factors, the type of lipid, and the presence of cholesterol do affect the extent to which LL-37 perturbs the lipids in the bilayer as observed with (31)P NMR. The (31)P spectra also show that micelles or other small, rapidly tumbling membrane fragments are not formed in the presence of LL-37, excluding a detergent-like mechanism. LL-37 does increase the lamellar to inverted hexagonal phase transition temperature of both PE model lipid systems and Escherichia coli lipids, demonstrating that it induces positive curvature strain in these environments. These results support a toroidal pore mechanism of lipid bilayer disruption by LL-37. PMID- 12767239 TI - Identification of the internal axial ligand of HO2-cobalt(III)-bleomycin: 1H[15N] HSQC NMR investigation of bleomycin, deglycobleomycin, and their hydroperoxide cobalt(III) complexes. AB - The identity of the axial ligand contributed by the drug in hydroperoxide-Co(III) bleomycin and hydroperoxide-Co(III)-deglycobleomycin has been in doubt. With each structure, a combination of (1)H[(15)N] HSQC and HMBC and (1)H COSY and NOESY NMR spectroscopy was used to observe and completely assign the nonaromatic (15)N chemical shifts of natural abundance bleomycin in the two hydroperoxide-Co(III) structures. Together with the (15)N assignments from a published 1D (15)N spectrum, the results permitted the assignment of the primary amine nitrogen to an axial ligand position in both structures. PMID- 12767240 TI - Kinetic mechanism of the GCN5-related chromosomal aminoglycoside acetyltransferase AAC(6')-Ii from Enterococcus faecium: evidence of dimer subunit cooperativity. AB - The aminoglycoside 6'-N-acetyltransferase AAC(6')-Ii from Enterococcus faecium is an important microbial resistance determinant and a member of the GCN5-related N acetyltransferase (GNAT) superfamily. We report here the further characterization of this enzyme in terms of the kinetic mechanism of acetyl transfer and identification of rate-contributing step(s) in catalysis, as well as investigations into the binding of both acetyl-CoA and aminoglycoside substrates to the AAC(6')-Ii dimer. Product and dead-end inhibition studies revealed that AAC(6')-Ii follows an ordered bi-bi ternary complex mechanism with acetyl-CoA binding first followed by antibiotic. Solvent viscosity studies demonstrated that aminoglycoside binding and product release govern the rate of acetyl transfer, as evidenced by changes in both the k(cat)/K(b) for aminoglycoside and k(cat), respectively, with increasing solvent viscosity. Solvent isotope effects were consistent with our viscosity studies that diffusion-controlled processes and not the chemical step were rate-limiting in drug modification. The patterns of partial and mixed inhibition observed during our mechanistic studies were followed up by investigating the possibility of subunit cooperativity in the AAC(6')-Ii dimer. Through the use of AAC-Trp(164) --> Ala, an active mutant which exists as a monomer in solution, the partial nature of the competitive inhibition observed in wild-type dead-end inhibition studies was alleviated. Isothermal titration calorimetry studies also indicated two nonequivalent antibiotic binding sites for the AAC(6')-Ii dimer but only one binding site for the Trp(164) --> Ala mutant. Taken together, these results demonstrate subunit cooperativity in the AAC(6')-Ii dimer, with possible relevance to other oligomeric members of the GNAT superfamily. PMID- 12767241 TI - Roles of individual amino acids in helix 14 of the membrane domain of proton translocating transhydrogenase from Escherichia coli as deduced from cysteine mutagenesis. AB - Proton-translocating nicotinamide nucleotide transhydrogenase is a membrane-bound protein composed of three domains: the hydrophilic NAD(H)-binding domain, the hydrophilic NADP(H)-binding domain, and the hydrophobic membrane domain. The latter harbors the proton channel. In Escherichia coli transhydrogenase, the membrane domain is composed of 13 transmembrane alpha helices, of which especially helices 13 and 14 contain conserved residues. To characterize the roles of the individual residues betaLeu240 to betaSer260 in helix 14, these were mutated as single mutants to cysteines in the cysteine-free background, and in the case of betaGly245, betaGly249, and betaGly252, also to leucines. In addition to the residues forming the helix, residues betaAsn238 and betaAsp239 were also mutated. Except for betaI242C, all mutants were normally expressed, purified, and characterized with respect to, e.g., catalytic activities and proton pumping. The results show that mutation of the conserved glycines betaGly245, betaGly249, and betaGly252, located on the same face of the helix, led to a general inhibition of all activities, especially in the case of betaGly252, suggesting a role of these glycines in helix-helix interactions. In contrast, mutation of the conserved serines betaSer250, betaSer251, and betaSer256 led to enhanced activities of all reactions, including the cyclic reaction which was mediated by bound NADP(H). Mutation of the remaining residues resulted in intermediate inhibitory effects. The results strongly support an important regulatory role of the membrane domain on the NADP(H)-binding site. PMID- 12767242 TI - Proton transport by proteorhodopsin requires that the retinal Schiff base counterion Asp-97 be anionic. AB - At pH >7, proteorhodopsin functions as an outward-directed proton pump in cell membranes, and Asp-97 and Glu-108, the homologues of the Asp-85 and Asp-96 in bacteriorhodopsin, are the proton acceptor and donor to the retinal Schiff base, respectively. It was reported, however [Friedrich, T. et al. (2002) J. Mol. Biol., 321, 821-838], that proteorhodopsin transports protons also at pH <7 where Asp-97 is protonated and in the direction reverse from that at higher pH. To explore the roles of Asp-97 and Glu-108 in the proposed pumping with variable vectoriality, we compared the photocycles of D97N and E108Q mutants, and the effects of azide on the photocycle of the E108Q mutant, at low and high pH. Unlike at high pH, at a pH low enough to protonate Asp-97 neither the mutations nor the effects of azide revealed evidence for the participation of the acidic residues in proton transfer, and as in the photocycle of the wild-type protein, no intermediate with unprotonated Schiff base accumulated. In view of these findings, and the doubts raised by absence of charge transfer after flash excitation at low pH, we revisited the question whether transport occurs at all under these conditions. In both oriented membrane fragments and liposomes reconstituted with proteorhodopsin, we found transport at high pH but not at low pH. Instead, proton transport activity followed the titration curve for Asp-97, with an apparent pK(a) of 7.1, and became zero at the pH where Asp-97 is fully protonated. PMID- 12767243 TI - Ketosynthases in the initiation and elongation modules of aromatic polyketide synthases have orthogonal acyl carrier protein specificity. AB - Many bacterial aromatic polyketides are synthesized by type II polyketide synthases (PKSs) which minimally consist of a ketosynthase-chain length factor (KS-CLF) heterodimer, an acyl carrier protein (ACP), and a malonyl-CoA:ACP transacylase (MAT). This minimal PKS initiates polyketide biosynthesis by decarboxylation of malonyl-ACP, which is catalyzed by the KS-CLF complex and leads to incorporation of an acetate starter unit. In non-acetate-primed PKSs, such as the frenolicin (fren) PKS and the R1128 PKS, decarboxylative priming is suppressed in favor of chain initiation with alternative acyl groups. Elucidation of these unusual priming pathways could lead to the engineered biosynthesis of polyketides containing novel starter units. Unique to some non-acetate-primed PKSs is a second catalytic module comprised of a dedicated homodimeric KS, an additional ACP, and a MAT. This initiation module is responsible for starter-unit selection and catalysis of the first chain elongation step. To elucidate the protein-protein recognition features of this dissociated multimodular PKS system, we expressed and purified two priming and two elongation KSs, a set of six ACPs from diverse sources, and a MAT. In the presence of the MAT, each ACP was labeled with malonyl-CoA rapidly. In the presence of a KS-CLF and MAT, all ACPs from minimal PKSs supported polyketide synthesis at comparable rates (k(cat) between 0.17 and 0.37 min(-1)), whereas PKS activity was attenuated by at least 50-fold in the presence of an ACP from an initiation module. In contrast, the opposite specificity pattern was observed with priming KSs: while ACPs from initiation modules were good substrates, ACPs from minimal PKSs were significantly poorer substrates. Our results show that KS-CLF and KSIII recognize orthogonal sets of ACPs, and the additional ACP is indispensable for the incorporation of non acetate primer units. Sequence alignments of the two classes of ACPs identified a tyrosine residue that is unique to priming ACPs. Site-directed mutagenesis of this amino acid in the initiation and elongation module ACPs of the R1128 PKS confirmed the importance of this residue in modulating interactions between KSs and ACPs. Our study provides new biochemical insights into unusual chain initiation mechanisms of bacterial aromatic PKSs. PMID- 12767244 TI - Characterization of the interaction of wheat HMGa with linear and four-way junction DNAs. AB - Wheat HMGa protein is a typical member of the plant HMGA family. It has four AT hooks and a histone H1-like region. A panel of deletion mutants of HMGa was generated to study the role of different regions of HMGa in its binding to 4H (a synthetic DNA that mimics the in vivo structure of intermediates of homologous recombination and DNA repair) and linear DNAs. Although the histone H1-like region of HMGa does not bind to 4H or linear DNAs, it does enhance the binding. Mutants with any two adjacent AT hooks show specific binding to both 4H and linear P268 (and P31) with different binding affinities, which is partly due to the flanking regions between AT hooks. Conformational studies indicate that the alpha-helical content of HMGa increases significantly when it binds to 4H compared to that after binding to P31, linear DNA. In contrast, linear DNA, but not 4H, undergoes substantial conformational change when it binds to HMGa, indicating that linear DNA is relatively more flexible than 4H. A more significant difference in the affinities of binding of the mutants of HMGa to 4H was observed compared to their affinities of binding to linear DNA, P31. These differences could be due to the rigidity of the DNA and the characters of the AT hook regions in the mutants. PMID- 12767245 TI - Structure of the ceramide moiety of GM1 ganglioside determines its occurrence in different detergent-resistant membrane domains in HL-60 cells. AB - To investigate the effect of the ceramide moiety of GM1 ganglioside on its association with detergent resistant membrane domains (DRMs) in human leukemia HL 60 cells, [(3)H] labeled GM1 molecular species (GM1s) with ceramides consisting of C18 sphingosine acetylated or acylated with C(8), C(12), C(14), C(16), C(18), C(22), C(24), C(18:1), C(22:1), or C(24:1) fatty acids (FAs), or C20 sphingosine acetylated or acylated with C(8) or C(18) FA were prepared and added to culture media. GM1s uptake by HL-60 cells was affected by the structure of their ceramides. Resistance to removal with trypsin and the stoichiometry of [(125)I] cholera toxin (CT) binding indicated that the added GM1s were incorporated into the membranes of the cells used for the isolation of DRMs in a manner resembling endogenous gangliosides. The ceramide moieties of the GM1s determined their occurrence in DRMs and the dependence of their recovery in this membrane fraction on the amount of Triton X-100 (TX) used for extraction as well as on cholesterol depletion. The GM1s with sphingosine acylated with C(14), C(16), C(18) C(22), or C(24) FAs were similarly abundant in DRMs. GM1s acylated with C(18:1), C(22:1), or C(24:1) were less abundant than those acylated with saturated FA of the same length. GM1s acetylated or acylated with C(8) FA were detected in DRMs in the lowest proportion. Depletion of 73% of cell cholesterol with methyl-beta cyclodextrin significantly affected the recovery in DRMs of GM1s acetylated or acylated with C(8) or unsaturated FAs but not of GM1 acylated with C(18), C(22), or C(24) FAs. After cross-linking with CT B subunit, all GM1s were recovered in DRMs in a similarly high proportion irrespective of their ceramide structure or cholesterol depletion. DRMs prepared with low TX concentration at the TX/cell protein ratio of 0.3:1 were separated by multistep sucrose density gradient centrifugation into two fractions. The GM1s with sphingosine acetylated or acylated with C(18) or C(18:1) FAs occurred in these fractions in different proportions. PMID- 12767246 TI - Comparative analysis of two different amide-to-ester bond mutations in the beta sheet of 4-oxalocrotonate tautomerase. AB - Here we describe the total chemical synthesis and biophysical characterization of two backbone-modified, ester bond-containing analogues of the homohexameric enzyme 4-oxalocrotonate tautomerase (4OT). The amide-to-ester bond mutations in the two analogues in this study, (OI2)4OT and (OI7)4OT, were designed to effectively delete specific backbone-backbone hydrogen bonds in the beta-sheet region of the native 4OT hexamer. The (OI2)4OT and (OI7)4OT analogues each contained one ester bond per monomer that effectively deleted 12 backbone backbone hydrogen bonds per hexamer. The structural properties of each analogue were characterized by size-exclusion chromatography (SEC), far-UV CD spectroscopy, and catalytic activity measurements, and they were found to be very similar to the structural properties of the wild-type enzyme. The results of equilibrium unfolding studies revealed that the (OI2)4OT and (OI7)4OT analogues were stabilized by 47.7 +/- 2.5 and 45.0 +/- 2.5 kcal/mol, respectively, under standard state conditions (1 M hexamer) as compared to a value of 69.6 +/- 3.3 kcal/mol for the wild-type control. Our results suggest that the two different, but structurally similar, backbone-backbone hydrogen bonds deleted in (OI2)4OT and (OI7)4OT make nearly equivalent contributions to the thermodynamic stability of the 4OT hexamer. PMID- 12767247 TI - Assuring quality in optometric practice. PMID- 12767248 TI - Practice standards for optometry and optometric practice accreditation. AB - BACKGROUND: To assist optometrists to deliver care more efficiently and effectively, in 1995 Optometrists Association Australia decided to develop standards that would assist optometrists in better managing their practices. Existing practice management standards for health professionals were thought to be either not specific enough for optometric practice or to have shortcomings in the context of optometric practice in Australia. METHODS: Following a literature search, material previously developed by Optometrists Association Australia to assist practitioners with management of their practices and standards from other professions were used to assist with the development of a draft set of standards for optometric practices in Australia. Successive drafts were circulated for comment to optometrists in practice, non-optometrists with experience in the development of practice standards for other health professions and to Australian General Practice Accreditation Limited. The comments were used to refine the standards and the accreditation guidelines to their final form. RESULTS: Optometric Practice Standards suitable for use in a practice accreditation program were developed. The standards comprise seven sections--Practice administration, Quality assurance, Rights and needs of the patient, Practice services, Practice facilities, Communication and Patient records. These sections are divided into criteria that provide the detail of the requirements of the standard. Indicators describing how criteria can be assessed accompany the criteria. PMID- 12767249 TI - Letter contrast sensitivity changes in early diabetic retinopathy. AB - PURPOSE: To investigate the discriminative ability of letter contrast sensitivity (CS) and visual acuity (VA) in detecting functional losses in participants with type 2 diabetes. METHODS: LogMAR VA and letter CS were measured on 20 type 2 diabetic and 24 age-matched control participants. The diabetic participants were sub-grouped according to the level of retinopathy present. RESULTS: Letter CS was able to distinguish those participants with and without macular oedema and those with no or minimal diabetic retinopathy from the control group, whereas VA was not. CONCLUSION: Letter CS may be used as an effective screening tool to assess damage to the visual pathway of diabetic participants both with and without clinically detectable signs. PMID- 12767250 TI - Image analysis and glare sensitivity in human age-related cataracts. AB - BACKGROUND: A new glare disability test and a diagnostic instrument, the Halometer, were proposed for measuring intraocular light scattering in the presence of human cataracts. The objectives of this work were to introduce new methods of computer-generated analysis of lens images and a glare disability test to validate the use of two new tests to measure the severity of cataract and to document and quantify changes in lens clarity at diagnosis in a group of patients with age-related cataract with minimal to advanced opacities. METHODS: The authors followed 28 patients (46 eyes) with an average age of 66.9 +/- 6 years. Tests included ophthalmoscopy, visual acuity, the halometer glare disability test, stereocinematographic slit-image and retro-illumination photography with subsequent interactive digital image analysis and three-dimensional (3-D) computer graphics of the areas of lens light scattering/absorbing. RESULTS: The intra-reader reproducibility of measuring techniques for cataractous changes was good. The Halometer instrument measures the angular distance from the glare source from which a specific target can be recognised. The source and the target are in the same vertical and tangential planes and to measure the angular distribution of the glare light, it is necessary to measure the incident light angle between the source and the target, by measuring the distance between the source and the target. The clinical Halometry readings assessed the index of the forward scattered light that reaches the retina and produces a veiling luminance (glare). The glare sensitivity score was determined from the angle of the glare source using both red and green optotypes, so that the effects of light absorption were separated from those of light scatter. DISCUSSION: The interactive and automated computerised system discriminated the photographic image features of the back light-scattering/absorb centres within different grey thresholds and documented objective lens findings such as optical density, areas of equidensities, geometric surfaces, opacity grading. Digital image analysis gave a topographic and 3-D assembling and visualisation for nuclear, cortical and posterior subcapsular opacities in human lenses revealed from the optical scanning tomographic study of the anterior eye segment. The results indicate on agreement between clinical findings and applied quantitative techniques of analysis. This system has the potential to be a useful objective clinical assessment of cataract. PMID- 12767251 TI - Repeatability of the C-100 colour vision test. AB - BACKGROUND: The C-100 colour vision test has been shown to have a high validity for diagnosing the type of red-green colour vision defect, however, there is little information on the repeatability of the test. This study examines the repeatability of the C-100 in classifying the colour vision defect as either protan or deutan. METHODS: The C-100 was administered on two occasions to 58 subjects with congenital red-green colour vision defects. The sessions were separated by a minimum period of 10 days. RESULTS: The repeatability of the C-100 was high with a kappa coefficient of agreement for diagnosis of 0.96. The few discrepancies were misclassifying protans as deutans. CONCLUSION: The C-100 is a highly repeatable test in terms of separating protans from deutans. However, if a discrepancy occurs, it is more likely to be a protan misclassified as a deutan rather than vice versa. PMID- 12767252 TI - The effect of observation time and contrast on visual acuity. AB - BACKGROUND: Field studies of the duration of fixation during daytime driving find that it is about 0.2 seconds on average. Thus, the time available to read traffic signs is often limited. There are other circumstances where time to read alphanumeric messages is limited. Data relating time of observation are needed for purposes of ergonomic design as the data available in the literature are limited and uncertain. METHOD: Visual acuity was measured using Landolt rings in negative contrast, which were presented on a computer screen with a background luminance of 32 cd/m(2) for durations that varied between two and 0.02 seconds at three different contrasts. RESULTS: Visual acuity depends on the log of the light dose reduced by a factor that may be related to absorption of the light by the eye media. For young observers, it was found that: VA = 0.57 (log C.t) + 1.705, where C is the contrast and is expressed as DL/L, where DL is the difference between target and background luminances in cd/m(2) and t is the observation time in seconds. CONCLUSION: This formula or the functions fitted to the data obtained can be used to calculate the increase in target size needed to allow for short observation times. PMID- 12767253 TI - The fundus in oculodermal melanocytosis. Is this a new observation? AB - An unusual case of bilateral oculodermal melanocytosis or the naevus of Ota is reviewed. In this condition, there is hyperpigmentation of ocular structures and the skin of the face. In this case, a pigmentary mottling of the fundus was observed using long wavelength light. PMID- 12767254 TI - Retinal angioma associated with von Hippel-Lindau disease. AB - Von Hippel-Lindau disease is a multi-system disorder that can produce hamartomas (benign tumour-like nodules) of the eyes, skin and nervous system. Retinal capillary angioma is a common ocular association of this congenital phakomatosis that may result in blinding sequelae, if not managed appropriately. We present a case of retinal angioma associated with von Hippel-Lindau disease and discuss the ocular and systemic signs, diagnosis and management. The optometrist is of particular importance in screening for this disorder, as it is often first detected in a routine eye examination. PMID- 12767255 TI - Characterization of the bipartite nuclear localization signal of protein LANA2 from Kaposi's sarcoma-associated herpesvirus. AB - LANA2 is a nuclear latent protein detected exclusively in Kaposi's sarcoma associated herpesvirus-infected B cells. The protein inhibits p53-dependent transactivation and apoptosis, suggesting an important role in the transforming activity of the virus. To explore the molecular mechanisms of its nuclear localization, fusion proteins of green fluorescent protein (EGFP) and deletion constructs of LANA2 were expressed in HeLa cells. Only the fragment comprising amino acid residues 355-440 of LANA2 localized in the cell nucleus. This fragment contains two closely located basic domains and forms a putative bipartite nuclear localization signal (NLS). The putative LANA2 NLS was able to target EGFP to the nucleus consistently. Site-directed mutation analyses demonstrated that LANA2 contains a functional bipartite NLS between amino acid positions 367 and 384. In addition, analysis of cells transfected with a cytoplasmic LANA2 mutant revealed that an appropriate subcellular localization may be crucial to regulate p53 activity. PMID- 12767256 TI - Heart failure: a hemodynamic disorder complicated by maladaptive proliferative responses. AB - Heart failure has traditionally been viewed as a hemodynamic syndrome characterized by fluid retention, high venous pressure, and low cardiac output. Over the past decade, however, it has become clear that because of deterioration and progressive dilatation (remodeling) of the diseased heart, this is also a rapidly fatal syndrome. The importance of prognosis came to be appreciated when clinical trials showed that therapy which initially improves such functional abnormalities, as high venous pressure and low cardiac output, often fail to improve survival, and that some drugs which improve hemodynamics worsen long-term prognosis. The latter is true for most vasodilators which, in spite of alleviating the adverse short-term consequences of high afterload, shorten survival. Notable exceptions are ACE inhibitors, whose vasodilator effects do not explain their ability to prolong survival; instead, these drugs slow both deterioration and remodeling of the failing heart. Inotropic agents, while providing immediate relief of symptoms, generally shorten long-term survival, whereas beta-blockers slow deterioration and remodeling, and reduce mortality. Aldosterone antagonists exert beneficial effects on prognosis that are not easily explained by their diuretic effects, but instead can be explained by their ability to inhibit signaling pathways that stimulate maladaptive hypertrophy, remodeling, apoptosis and other deleterious responses that cause deterioration of the failing heart. These and other findings demonstrate that heart failure is more than a hemodynamic disorder; these patients suffer from maladaptive proliferative responses that cause cardiac cell death and progressive dilatation that play a key role in determining the poor prognosis in this syndrome. PMID- 12767257 TI - Tetrathiomolybdate anticopper therapy for Wilson's disease inhibits angiogenesis, fibrosis and inflammation. AB - The need for agents to lower body copper in Wilson's disease, a disease which results from copper toxicity has been the driving force for the development of the effective anticopper drugs penicillamine, trientine, zinc, and now tetrathiomolybdate (TM). Because of its rapid action, potency, and safety, TM is proving to be a very effective drug for initial treatment of acutely ill Wilson's disease patients. Beyond this, TM has antiangiogenic effects, because many proangiogenic cytokines require normal levels of copper. This has led to use of TM in cancer, where it is generally effective in animal tumor models, and has shown efficacy in preliminary clinical studies. Most recently, it has been found that TM has antifibrotic and antiinflammatory effects through inhibition of profibrotic and proinflammatory cytokines. PMID- 12767258 TI - Physiological and pathological roles of Apaf1 and the apoptosome. AB - Different cellular pathways can lead to apoptosis. Apaf1 is the molecular core of the apoptosome, a multiproteic complex mediating the so-called mitochondrial pathway of cell death. The importance of this pathway during development has been clearly demonstrated by knocking out key genes. Also, the relevance of Apaf1 dosage during development has been recently underlined. Moreover, a growing body of evidences seems to point out a possible role of the mitochondria-dependent apoptosis in different pathologies. In particular, we discuss here some recent evidences regarding the putative role of the apoptosome in neurodegeneration and cancer. PMID- 12767259 TI - RNA-binding ability of PIPPin requires the entire protein. AB - Post-transcriptional fate of eukaryotic mRNAs depends on association with different classes of RNA-binding proteins (RBPs). Among these proteins, the cold shock domain (CSD)-containing proteins, also called Y-box proteins, play a key role in controlling the recruitment of mRNA to the translational machinery, in response to environmental cues, both in development and in differentiated cells. We recently cloned a rat cDNA encoding a new CSD-protein that we called PIPPin. This protein also contains two putative double-stranded RNA-binding motifs (PIP(1) and PIP(2)) flanking the central CSD, and is able to bind mRNAs encoding H1 degrees and H3.3 histone variants. In order to clarify the role of each domain in the RNA-binding activity of PIPPin, we constructed a number of different recombinant vectors, encoding different regions of the protein. Here we report that only recombinant proteins that contain all the putative PIPPin domains show RNA-binding ability. PMID- 12767260 TI - Screening for microdeletions in human Y chromosome--AZF candidate genes and male infertility. AB - About 30% of couple infertilities are of male origin, some of them caused by genetic abnormalities of the Y chromosome. Deletions in AZF region can cause severe spermatogenic defects ranging from non-obstructive azoospermia to oligospermia. The intracytoplasmatic sperm injection technique (ICSI) is rapidly becoming a versatile procedure for human assisted reproduction in case of male infertility. The use of ICSI allows Y chromosome defects to be passed from father. The goal of our study is to evaluate the frequency of microdeletions in the long arm of Y chromosome, within the AZF regions, in these cases of infertilities, using molecular genetics techniques. Thirty infertile men with azoospermia or oligozoospermia, determined by spermogram, were studied after exclusion of patients with endocrine or obstructive causes of infertility. Peripheral blood DNA was extracted from each patient, then amplified by multiplex PCR with STS genomic markers from the Y chromosome AZF zones. Each case was checked by multiplex PCR through coamplification with the SRY marker. Three men with microdeletions of the long arm of the Y chromosome were diagnosed among the 30 patients, corresponding to a proportion of 10%. The relatively high proportion of microdeletions found in our population suggest the need for strict patient selection to avoid unnecessary screening for long arm Y chromosome microdeletions. The molecular diagnostics was performed according to the current European Academy of Andrology laboratory guidelines for molecular diagnosis of Y chromosomal microdeletions. PMID- 12767261 TI - Glucose deprivation induces mitochondrial dysfunction and oxidative stress in PC12 cell line. AB - Glucose metabolism plays a pivotal role in many physiological and pathological conditions. To investigate the effect of hypoglycemia (obtained by glucose deprivation) on PC12 cell line, we analyzed the cell viability, mitochondrial function (assessed by MTT reduction, cellular ATP level, mitochondrial transmembrane potential), and the level of reactive oxygen species (ROS) after glucose deprivation (GD). Upon exposure to GD, ROS level increased and MTT reduction decreased immediately, intracellular ATP level increased in the first 3 hours, followed by progressive decrease till the end of GD treatment, and the mitochondrial transmembrane potential (deltapsi(m)) dropped after 6 hours. Both necrosis and apoptosis occurred apparently after 24 hours which was determined by nuclei staining with propidium iodide(PI) and Hoechst 33342. These data suggested that cytotoxicity of GD is mainly due to ROS accumulation and ATP depletion in PC12 cells. PMID- 12767262 TI - Phospholipase A2 modulates respiratory burst developed by neutrophils in patients with rheumatoid arthritis. AB - Activated by bacterial peptides, phorbol esters, calcium ionophores and other agonists, neutrophils (PMNs) release the proinflammatory mediator, arachidonic acid (AA) via the intervention of phospholipase A(2) (PLA(2)). AA may play an essential role in activation of NADPH-oxidase, which is involved in the generation of superoxide anion by neutrophils. The present study is focused on the involvement of PLA(2) in the respiratory burst developed by PMNs isolated from patients with rheumatoid arthritis (RA). PLA(2) exists in very high levels in diseases such as rheumatoid arthritis and may cause acute inflammatory and proliferative changes in synovial structures. The respiratory burst was evaluated as superoxide anion release, using an amplified chemiluminescence method. The assays were performed using PMNs untreated or treated with different doses of stimulatory reagents (phorbol 12-myristate-13-acetate (PMA), calcium ionophore (A23187)). Our data suggested that PMA stimulated the production of superoxide anion in a dose-response manner, as compared with A23187, which did not induce a significant release of superoxide anion in PMNs-RA. The exogenous addition of AA significantly amplified the superoxide anion release by PMNs-RA stimulated with PMA and to a lesser extent, by PMNs stimulated with A23187. AA has also reversed the inhibitory effect of arachidonyl-trifluorometylketone and E-6 (bromomethylene)tetrahydro-3-(1-naphthalenyl)2H-pyran-2-one (BEL) on the superoxide anion release by PMNs-RA. In conclusion, the differential responses to these two agents suggested that different isoforms of PLA(2) were activated by A23187 or PMA, and support the idea that activation of these different PLA(2) served distinct functions of PMNs. Therefore, the inhibition of PLA(2) enzymes might be of great importance in the immunotherapy of rheumatoid arthritis. PMID- 12767263 TI - Expression and significance of new tumor suppressor gene PTEN in primary liver cancer. AB - OBJECTIVE: To investigate expression and significance of PTEN gene in primary hepatocellular carcinoma (HCC). METHODS: Immunohistochemical peroxidase conjugated streptavidin (SP) method was used to detect expression of PTEN gene in 120 cases of primary HCC and its adjacent tissue 10 cases of normal liver tissue. The relationship between expression of tumor suppressor gene of PTEN and the percentage of lymph node metastasis of HCC was analyzed. RESULTS: It was shown that PTEN gene was expressed in all 10 cases of normal liver tissues and paracancerous liver tissues. The staining was localized mainly in cytoplasm. Expression of PTEN in 120 cases of HCC were as follows: 12.5% were negative, 17.5% were weak positive, and 70% were strong positive. At time of diagnosis, 33/120 (27.5%) presented lymph node metastasis. Lymph node metastases were present in 80% (12 out of 15) PTEN negative HCC, 57.14% (12 out of 21) PTEN weak positive HCC and only 10.71% (9 out of 84) PTEN intense positive HCC, (P<0.05). Therefore, PTEN tumor suppressor gene malfunction seems to be involved in metastasing capacity of HCC. CONCLUSION: This study suggests that PTEN gene was deleted or weakly expressed in primary hepatocellular carcinoma, which is probably related to its tumorigenesis. PMID- 12767264 TI - Plasma membrane potential interferes with the respiratory burst of peripheral granulocytes. AB - Membrane potential is involved in the regulation of several immune functions developed by granulocytes. The Na(+)/K(+) gradient across the plasma membrane, mainly generated by the Na(+)/K(+) pump, plays a key role in the maintenance of membrane potential. This study is focused on the correlation between plasma membrane potential and the in vitro receptor - triggered respiratory burst of normal human peripheral granulocytes. The respiratory burst was measured as superoxide anion release by the cytochrome c reduction test and plasma membrane potential was modulated by experimental changes of the extracellular potassium concentration. Results show a differentiated cellular response, depending on the in vivo activation state and on the signals received in vitro by granulocytes via CR3 or FcgammaR. Alteration of the membrane potassium gradient modulates the respiratory burst of unstimulated and CR3-activated cells, whilst it does not seem to significantly interfere with the signals delivered by FcgammaR. PMID- 12767265 TI - Imbalance of peripheral B lymphocytes and NK cells in rheumatoid arthritis. AB - The study was focused on several cellular immune disorders correlated with the imbalance between peripheral blood B lymphocytes and NK cells in severe rheumatoid arthritis. By flow cytometry we calculated the proportions of T, T helper, T cytotoxic/suppressor, B lymphocytes and natural killer cells in peripheral blood. The mitogen-induced proliferation of peripheral lymphocytes was measured by tritium-labeled uridine incorporation. Experimental data highlight a connection between annomal values of the B to natural killer cells ratio and disorders of the peripheral mononuclear cells concentration. We also showed that the polyclonal proliferation capacity of peripheral lymphocytes in rheumatoid arthritis is solely related to the B to natural killer cells ratio or to the natural killer cells proportion. The study reveals a potential role of the imbalance between proportions of peripheral B lymphocytes and natural killer cells in the immune pathogenesis of rheumatoid arthritis, thus pointing out an interrelation between the adaptive and innate immune systems. PMID- 12767266 TI - Plasma 249Ser p53 mutation in patients with hepatocellular carcinoma residing in a high risk area. AB - OBJECTIVE: To investigate plasma p53 mutation in hepatocellular carcinoma (HCC) patients from Qidong and to define its significance. METHODS: Blood samples from 25 hepatocellular carcinoma patients, 20 cirrhotic patients and 30 healthy controls in Qidong area. DNA was extracted and purified from 200 microl of plasma from each sample. The 249(Ser) p53 mutation was detected by restriction digestion analysis and by direct sequencing of exon-7 PCR products. RESULTS: G-->T transversion at the third base of 249 codon resulting in 249(Arg)-->249(Ser) mutation in exon 7 of p53 gene were found in 11/25(44%) hepatocellular carcinoma cases, 4/20 (20%) cirrhotics, and 2/30 (7%) healthy controls (p<0.01). CONCLUSIONS: These data show that the 249(Ser) p53 mutation in plasma is strongly associated with hepatocellular carcinoma patients in Qidong area and the mutation should be screened as a new early diagnostic marker for HCC. PMID- 12767267 TI - From fetal neuroblastic nests to adult neuronal glomeruli in human cervical sympathetic ganglia. PMID- 12767274 TI - Secretion of endogenous kallikreins 2 and 3 by androgen receptor-transfected PC-3 prostate cancer cells. AB - Androgen independent PC-3 cells lack androgen receptor (AR) expression and do not produce kallikrein 2 (hK2) or 3 (prostate-specific antigen, PSA). In this paper, we examined the ability of androgens to stimulate PSA and hK2 production in AR transfected PC-3 cells (PC-3(AR)) and compared this to LNCaP cells. PSA and hK2 were measured in the culture medium and cell lysates using an ELISA-based immunofluorometric assay. Only androgens were able to induce PSA and hK2 secretion in PC-3(AR) cells in a dose- and time-dependent manner depending on the level of AR present. The level of androgen-induced PSA and hK2 secretion in PC 3(AR) cells was approximately 1.5 and 0.9% that induced in LNCaP cells, respectively. Insulin-like growth factor-I (IGF-I), which has been shown to activate AR in the absence of ligand, did not activate PSA secretion in the absence of androgen, but further increased the dihydrotestosterone-induced PSA secretion in PC-3(AR) cells. The lack of PSA and hK2 production in parental PC-3 cells is thus a result of their lack of AR expression. PSA and/or hK2 production in PC-3(AR) cells can thus serve as an endogenous reporter system to investigate AR action or to screen putative endocrine disrupters. PMID- 12767275 TI - Pharmacological profile of MEN 11066, a novel potent and selective aromatase inhibitor. AB - MEN 11066 is a new non-steroidal compound which potently inhibits human placenta (K(i)=0.5 nM) and rat ovarian (K(i)=0.2 nM) aromatase in vitro. In vivo, a single oral dose of 0.3 mgkg(-1) significantly decreased uterus weight in immature rats after stimulation of uterus growth by androstenedione. MEN 11066 reduced in a dose-dependent manner plasma estradiol levels in adult female rats treated with pregnant mare serum gonadotropin (PMSG). After 2 weeks of repeated daily treatment in adult rats, a significant decrease in uterine weight was observed together with a 65% decrease in plasma estradiol, whereas plasma levels of testosterone, progesterone, aldosterone, corticosterone, cholesterol, LH and FSH were not affected. The lack of any effect by MEN 11066 on adrenal steroids was confirmed by the unchanged plasma corticosterone and aldosterone levels in immature rats and also in adult rats when the repeated treatment with MEN 11066 (15 days) was followed by the administration of a synthetic ACTH analogue. No change in 11beta-hydroxylase or 21-hydroxylase activities was produced in vitro by the addition of 10 microM MEN 11066. Fifteen-day treatment with MEN 11066 did not produce changes in several rat hepatic enzymatic activities involved in the metabolism of xenobiotics. These results demonstrated that MEN 11066 is a potent inhibitor of aromatase which does not interfere with the cytochrome P450 involved in the synthesis of other steroids or in the metabolism of xenobiotics. PMID- 12767276 TI - Endometrial effects of selective estrogen receptor modulators (SERMs) on estradiol-responsive gene expression are gene and cell-specific. AB - Three selective estrogen receptor modulator (SERM) drugs which included 4-OH tamoxifen (Tam), EM-800 (EM) and GW 5638 (GW) were investigated to determine their ability to inhibit estradiol-responsive gene expression in sheep endometrium. The uteri of ovariectomized ewes (10 ewes per SERM group) were infused with 10(-7)M SERMs for 24h prior to hysterectomy. Five ewes from each group received 50 microg 17beta-estradiol (E2) and the remaining five ewes received vehicle 18 h prior to hysterectomy. Northern blot analyses and in situ hybridization demonstrated that E2 treatment increased estrogen receptor (ER), progesterone receptor (PR), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), and cyclophilin (CYC) mRNA levels in most endometrial cells examined. Tam and GW exhibited characteristics similar to E2 by increasing ER gene expression, but they antagonized the E2-induced increases in PR and CYC mRNA levels. EM acted as an E2-agonist of GAPDH gene expression, but antagonized the E2 up-regulation of ER, PR and CYC gene expression in most endometrial cells. Immunohistochemistry determined that EM decreased ER protein levels in the glandular epithelium, and the SERMs investigated antagonized increases in PR protein levels in endometrium. In conclusion, GW and EM exhibit fewer agonist effects than Tam on endometrial gene expression. EM demonstrated the greatest antagonism of E2-enhanced levels of ER, PR and CYC, likely due to the inhibition of ER gene expression at both mRNA and protein levels. PMID- 12767277 TI - Myometrial effects of selective estrogen receptor modulators on estradiol responsive gene expression are gene and cell-specific. AB - We examined in vivo effects of selective estrogen receptor modulators (SERMs) 4 OH-tamoxifen (Tam), GW 5638 (GW) and EM-800 (EM) on myometrial gene expression. The uteri of ovariectomized ewes were infused with 10(-7)M of one SERM via indwelling catheters for 24h preceding hysterectomy. Half of the ewes in each SERM group received an intramuscular injection of 50 microg 17beta-estradiol (E2) 18 h prior to hysterectomy. Northern blot analysis and in situ hybridization demonstrated that E2 increased estrogen receptor (ER), progesterone receptor (PR) and cyclophilin (CYC) gene expression in the cells of both inner layer of myometrium (IM) and outer layer of myometrium (OM) as well as glyceraldehyde 3 phosphate dehydrogenase (GAPDH) gene expression in OM. Tam also increased ER mRNA levels in OM. EM appeared to increase ER gene expression, but antagonized E2's up regulation of PR and CYC gene expression in both IM and OM. Tam and GW also antagonized E2 up-regulation of PR gene expression in OM but not IM. No SERM affected GAPDH gene expression with or without E2. Immunohistochemistry indicated that E2 increased nuclear ER and PR protein levels in both IM and OM. EM was unique in up-regulating ER protein levels, opposite to its effects in endometrial cells. All SERMs tested antagonized this increase in PR immunostaining preferentially in OM compared to the IM layer. These results illustrate gene and cell layer-specific effects of SERMs in sheep myometrium. PMID- 12767278 TI - In vitro and in vivo models for the evaluation of potent inhibitors of male rat 17alpha-hydroxylase/C17,20-lyase. AB - The C(17,20)-lyase is a key enzyme in the biosynthesis of androgens by both the testes and adrenals. A complete inhibition of this enzyme would provide an alternative means of androgen suppression for the treatment of prostatic cancers. In the present study, the inhibitory effects of new non-steroidal compounds were tested in vitro on rat C(17,20)-lyase versus abiraterone, a reference steroidal inhibitor. Their activities were also evaluated in vivo on plasma testosterone (T) and luteinizing hormone (LH) levels and on testes, adrenals, seminal vesicles (SV) and ventral prostate (VP) weights after 3 days of oral treatment to adult male rats (50mg/kg per day p.o.). Inhibition in the nanomolar range was obtained with TX 977, the lead racemate product in this series, and optimization is ongoing based on a slight dissociation observed between its two diastereoisomers, TX 1196-11 (S) and TX 1197-11 (R). These non-steroidal compounds (including YM 55208, a reference competitor) proved to be more active in vivo than abiraterone acetate in this model, but the observed impact on adrenal weight suggests that the specificity of lyase inhibition versus corticosteroid biosynthesis deserves further investigations with this new class of potentially useful agents for the treatment of androgen-dependent prostate cancer. PMID- 12767279 TI - Transcriptional inhibition of the human insulin receptor gene by aldosterone. AB - In earlier studies, we reported reduced human insulin receptor (hIR) mRNA levels, insulin binding and insulin responsiveness in U-937 human promonocytic cells treated with aldosterone. The mechanism for this inhibition could be diminished IR gene transcription, since aldosterone did not affect hIR mRNA stability. All the effects were mediated by a downregulation of the mineralocorticoid receptor (MR, NR3C2) expressed at both the RNA and protein levels, suggesting that MR could act as a transcription factor that binds to hormone response elements in the hIR gene promoter. Indeed, MR has been shown to bind glucocorticoid response elements (GREs) in target genes. Given that five GREs have been characterized in the hIR promoter, we decided to test whether these elements could mediate the aldosterone-elicited inhibition of hIR expression detected by us in U-937 cells. In the present report, we demonstrate that aldosterone inhibits the activity of the hIR wild-type promoter by 23%, and causes 23 and 31% reductions in the activity of progressive deletions of this promoter comprised of fragments up to 1473 and -876bp, respectively. This indicates that the -876 to -271bp region of the hIR promoter may be sufficient for this transcriptional inhibition by aldosterone. We also provide evidence for direct MR interaction with some of the GREs of this promoter region, specifically with the cGRE1 and cGRE3, presumably as MR-MR homodimers, and with pGRE as a MR-GR heterodimer. This heterodimer may play the most relevant role and participate in the cross-talk between mineralocorticoids, glucocorticoids and insulin signalling in U-937 cells. PMID- 12767280 TI - Effects of novel 17alpha-hydroxylase/C17, 20-lyase (P450 17, CYP 17) inhibitors on androgen biosynthesis in vitro and in vivo. AB - Aiming at the development of new drugs for the treatment of prostate cancer, the effects of steroidal compounds and one non-steroidal substance on androgen biosynthesis were evaluated in vitro and in vivo. Sa 40 [17-(5-pyrimidyl)androsta 5,16-diene-3beta-ol], its 3-acetyl derivate Sa 41 and BW 19 [3,4-dihydro-2-(4 imidazolylmethyl)-6-methoxy-1-methyl-naphthalene] are compounds from our group, which have been developed as inhibitors of CYP 17 (17alpha-hydroxylase-C17, 20 lyase, the key enzyme in androgen biosynthesis). They have been compared with CB 7598 [abiraterone: 17-(3-pyridyl)androsta-5,16-diene-3beta-ol], its 3-acetyl compound CB 7630 and ketoconazole, compounds which already have been used clinically. The most potent compound toward human CYP 17 (testicular microsomes) was Sa 40 (IC(50) value of 24 nM), followed by Sa 41, CB 7598, BW 19, CB 7630 and ketoconazole. Sa 40 shows a type II difference spectrum and a non-competitive type of inhibition (K(i) value of 16 nM). No recovery of enzyme activity was observed after preincubation of CYP 17 with Sa 40 and subsequent charcoal treatment. In Escherichia coli cells coexpressing human CYP 17 and NADPH-P450 reductase, Sa 40 was more active than CB 7598 and BW 19, whereas the acetyl compounds were not active. The latter three compounds were equally active towards rat CYP 17. Male Sprague-Dawley (SD) rats were administered daily for 14 days BW 19 and the acetyl derivatives Sa 41 and CB 7630 as prodrugs (0.1 mmol/kg intraperitoneally). The test compounds strongly reduced plasma testosterone concentration, as well as prostate and seminal vesicles weights. They showed moderate inhibitory effects on the weights of levator ani, bulbocavernosus and testes, whereas they led to an increase in adrenal and pituitary weights. The only exception was BW 19 which did not change pituitary weights. Based on its superiority on the human enzyme, it was concluded that Sa 40 in its 3beta-acetate form (Sa 41) could be a promising candidate for clinical evaluation. PMID- 12767281 TI - Plasma 11beta-hydroxy-4-androstene-3,17-dione: comparison of a time-resolved fluoroimmunoassay using a biotinylated tracer with a radioimmunoassay using a tritiated tracer. AB - The plasma concentration of 11beta-hydroxy-4-androstene-3,17-dione (11beta) is very high in 21-hydroxylase deficiency, Cushing's syndrome, and hyperandrogenism of adrenal origin, and very low in congenital 11-hydroxylase deficiency and adrenal insufficiency. Thus, when plasma 4-androstenedione is elevated, it is useful to measure the plasma 11beta level in order to determine the adrenal or ovarian origin of the hyperandrogenism. To eliminate disadvantages related to the 11beta radioimmunoassay (RIA), which uses a tritiated tracer, as well as the high cost associated with scintillation proximity assay (SPA), we developed a non isotopic 11beta assay that utilizes an 11beta-biotin conjugate synthesized in our laboratory to measure time-resolved fluorescence after addition of streptavidin europium to microtitration wells. The analytical qualities of this assay are very similar to those of the radioimmunoassay using a tritiated tracer, and an extraction step followed by celite chromatography (which separates 11beta from interfering plasma steroids) prior to a final radioimmuno-competition step. The correlation coefficient between 11beta levels measured by time-resolved plasma 11beta fluoroimmunoassay (TR-FIA) and RIA was 0.965.Finally, the TR-FIA technique was more sensitive and of greater precision than the RIA method. PMID- 12767282 TI - Dithioerythritol (DTE) prevents inhibitory effects of triphenyltin (TPT) on the key enzymes of the human sex steroid hormone metabolism. AB - Organotins are known to induce imposex (pseudohermaphroditism) in marine neogastropods and are suggested to act as specific endocrine disruptors, inhibiting the enzyme-mediated conversion of steroid hormones. Therefore, we investigated the in vitro effects of triphenyltin (TPT) on human 5alpha-reductase type 2 (5alpha-Re 2), cytochrome P450 aromatase (P450arom), 17beta-hydroxysteroid dehydrogenase type 3 (17beta-HSD 3), 3beta-HSD type 2 and 17beta-HSD type 1 activity. First, the present study demonstrates that significant amounts of TPT occurred in the blood of eight human volunteers (0.17-0.67 microg organotin cation/l, i.e. 0.49-1.92 nmolcation/l). Second, TPT showed variable inhibitory effects on all the enzymes investigated. The mean IC(50) values were 0.95 microM for 5alpha-Re 2 (mean of n=4 experiments), 1.5 microM for P450arom (n=5), 4.0 microM for 3beta-HSD 2 (n=1), 4.2 microM for 17beta-HSD 3 (n=3) and 10.5 microM for 17beta-HSD 1 (n=3). To exclude the possibility that the impacts of TPT are mediated by oxidizing essential thiol residues of the enzymes, the putative compensatory effects of the reducing agent dithioerythritol (DTE) were investigated. Co-incubation with DTE (n=3) resulted in dose-response prevention of the inhibitory effects of 100 microM deleterious TPT concentrations on 17beta HSD 3 (EC(50) value of 12.9 mM; mean of n=3 experiments), 3beta-HSD 2 (0.90 mM; n=3), P450 arom (0.91 mM; n=3) and 17beta-HSD 1 (0.21 mM; n=3) activity. With these enzymes, the use of 10mM DTE resulted in an at least 80% antagonistic effect, whereas, the effect of TPT on 5alpha-Re 2 was not compensated. In conclusion, the present study shows that TPT acts as an unspecific, but significant inhibitor of human sex steroid hormone metabolism and suggests that the inhibitory effects are mediated by the interaction of TPT with critical cysteine residues of the enzymes. PMID- 12767283 TI - Time-resolved fluoroimmunoassay for equol in plasma and urine. AB - We present a method for the determination of the isoflavan equol in plasma and urine. This estrogenic isoflavan, which is formed by the action of the intestinal microflora, may have higher biological activity than its precursor daidzein. High urinary excretion of equol has been suggested to be associated with a reduction in breast cancer risk. The method is based on time-resolved fluoroimmunoassay, using a europium chelate as a label. After synthesis of 4'-O-carboxymethylequol the compound is coupled to bovine serum albumin (BSA), then used as antigen to immunize rabbits. The tracer with the europium chelate is synthesized using the same 4'-O-derivative of equol. After enzymatic hydrolysis (urine) or enzymatic hydrolysis and ether extraction (plasma) the immunoassay is carried out. The antiserum cross-reacted to variable extent with some isoflavonoids. For the plasma method the cross-reactivity does not seem to influence the results, which were highly specific. The overestimation of the values using the urine method (164%) compared to the results obtained by a gas chromatography-mass spectrometry (GC-MS) method is probably due to some influence of the matrix on the signal, and interference of structurally related compounds. It is suggested that plasma assays are used but if urine samples are measured a formula has to be used to correct the values making them comparable to the GC-MS results. The correlation coefficients between the time-resolved fluoroimmunoassay (TR-FIA) methods and GC MS methods were high; r-values for the plasma and urine method, were 0.98 and 0.91, respectively. The intra-assay coefficient of variation (CV%) for the TR-FIA plasma and urine results at three different concentrations vary between 5.5-6.5 and 3.4-6.9, respectively. The inter-assay CV% varies between 5.4-9.7 and 7.4 7.7, respectively. The working ranges of the plasma and urine assay are 1.27-512 and 1.9-512nmol/l, respectively. PMID- 12767284 TI - Pollutant removal from municipal sewage lagoon effluents with a free-surface wetland. AB - This research project was initiated to refine the knowledge available on the treatment of rural municipal wastewater by constructed wetlands. To determine the treatment capacity of a constructed wetland system receiving municipal lagoon effluents, the wetland was monitored over one treatment season, from May 19 to November 3, 2000. The wetland system consisted of a three-cell free-surface wetland, phosphorus adsorption slag filters and a vegetated filter strip. Bimonthly water samples at the inlet and outlet of each component of the wetland system were analysed for biochemical oxygen demand, nitrate and nitrite, ammonia and ammonium, total Kjeldahl nitrogen (TKN), total suspended solids (TSS), total phosphorus (TP), ortho-phosphate (ortho-PO(4)), fecal coliforms (FCs) and Escherichia coli. The free-surface wetland cells treating the lagoon effluents achieved removals as follows: biochemical oxygen demand (34%), ammonia and ammonium (52%), TKN (37%), TSS (93%), TP (90%), ortho-PO(4) (82%), FCs (52%) and E. coli (58%). The wetland cells reduced total nitrogen, TP and biochemical oxygen demand to levels below the maximum permissible levels required for direct discharge to nearby receiving waters (TN<3.0 mg x L(-1), TP<0.3 mg x L(-1), BOD(5)<3.0 mg x L(-1)). The vegetated filter strip treating the effluents from the wetland cells achieved removals as follows: biochemical oxygen demand (18%), ammonia and ammonium (28%), TKN (11%), TSS (22%), TP (5%), FCs (28%) and E. coli (22%). It may therefore serve as an additional treatment stage further reducing the concentrations of these mentioned parameters. The slag filters reduced TP in the lagoon effluents by up to 99%, and, in this study, were concluded to be effective phosphorus adsorbers. PMID- 12767285 TI - Removal of resin acids and sterols from pulp mill effluents by activated sludge treatment. AB - The wastewater treatment plant of an elemental chlorine free bleaching kraft pulp mill located in eastern Finland was sampled in order to study the fate of wood extractives and the toxicity to luminescence bacteria (Vibrio fischeri) in different parts of the plant. Resin acids and sterols were analyzed from water, particles and sludge samples during three different runs. Waters before biotreatment and primary sludge were found to be toxic; but in the activated sludge treatment toxicity was removed. During wastewater treatment, concentrations of wood extractives were reduced over 97%. In activated sludge treatment, over 94% of the resin acids and over 41% of the sterols were degraded or transformed to other compounds. Furthermore, in general, less than 5% of the resin acids and over 31% of the sterols were removed in biosludge to the sludge thickener. Most of the extractives were discharged attached to particles. Although some disturbing factors increased the load of wood extractives during samplings, these factors did not affect the operational efficiency of the secondary treatment system. PMID- 12767286 TI - A method for assessing the effect of water quality changes on plumbosolvency using random daytime sampling. AB - The Mann-Whitney U-test is used to demonstrate the impact of phosphate on lead concentrations measured at customer properties. This test is statistically robust and particularly efficient for the type of distributions encountered in lead random daytime sampling. This non-parametric technique is developed to provide a best estimate of the lead reduction that results from a change in plumbosolvency conditions. The method is illustrated with compliance data collected before and after the introduction of phosphate at customer properties in the north west of England. Limitations due to operational factors are highlighted. Using a Monte Carlo simulation of the variability of lead random daytime samples it is shown that the method might be practical when assessing the impact of incremental changes in phosphate concentration on plumbosolvency. PMID- 12767287 TI - A model of microbial activity in lake sediments in response to periodic water column mixing. AB - Under stagnant conditions, the mass transport of a soluble substrate from a lake's water column to the sediment/water interface is limited by molecular diffusion. Stagnant conditions coupled with a continuing sediment biological demand create a substrate depletion zone above the sediment/water interface. The frequency at which the substrate depletion zone is destroyed by internal seiches and other intermittent flow phenomena influences the time-averaged substrate concentration at the sediment/water interface. A more frequent mixing results in a greater time-averaged interface concentration and consequently affects the amount of microbial biomass that can be supported in the lake sediments and the flux of the substrate into the sediment. A one-dimensional, two-substrate model is used to examine the impact of mixing frequency on the activity of sulfate reducing bacteria (SRB) in lake sediments. In the model, sulfate is supplied from the water column, while acetate is generated within the sediments. Mass transport to and within the sediments is by molecular diffusion except for instantaneous mixing events. Between mixing events, sulfate concentration gradients form above the sediment/water interface in the diffusive boundary layer. Sulfate depletion zones can be centimeters thick. When typical biological rate and diffusion coefficients for sulfate and acetate are used as inputs, the model indicates that a more frequent water-column mixing results in greater SRB concentrations. For an assumed bulk water-column sulfate concentration of 4.8 mg x l(-1), the sediment SRB concentrations for the modeled hourly, 6-hourly, daily, and weekly mixing frequencies were 175, 136, 91, and 30 mg x m(-2), respectively. The model also predicts higher time-averaged sulfate flux rates at more frequent water-column mixing. The time-averaged sulfate flux rates for the hourly, 6-hourly, daily, and weekly mixing frequencies were 1.26, 1.13, 0.78, and 0.30 mg x m(-2)h(-1), respectively. Thus, mixing frequency can significantly impact microbial activity in lake sediments. PMID- 12767288 TI - Characterisation of bed sediments and suspension of the river Po (Italy) during normal and high flow conditions. AB - Grain-size distribution, major elements, nutrients and trace metals were determined in bed sediments and suspension collected at 10 representative sites along the river Po under normal and high flow conditions. Grain-size distribution and major element composition of suspension highlighted the presence of two distinct particle populations in the upper-middle Po (coarser particles, lower carbonate content) and in the lower Po (finer particles, higher carbonate content). This change partly reflects the geological differences between the two parts of the basin, and also the presence of a hydroelectric power plant at Isola Serafini (Piacenza). With respect to environmental quality issues, bed sediments and suspension provide similar results. A moderate nutrient pollution is found in all but the uppermost parts of the river basin, while the most significant inputs of trace metals appear to originate from the urban areas of Turin and Milan. Calculation of sediment enrichment factors identifies Cd, Cu, Hg and Zn as the most impacted elements by human activities. On the other hand, the high levels of Ni and Cr throughout the river seem to derive mainly from the presence of basic rocks in the upper and middle parts of the basin. Both nutrient and trace metal particulate concentrations substantially decrease under high flow conditions possibly due to "flushing" of contaminated bed sediments and resuspension of coarser material. Under normal flow conditions, water hydrochemistry and concentrations of some elements (As, Ca, Cr, Cu, K, Mg, Mn, Na, Ni, and Pb) in the dissolved phase (<0.45 microm) were also determined. Calculation of trace metals partition coefficients shows that the relative importance of the particulate and water phases varies in response to water hydrochemistry and suspended solid content, but that most elements achieve a conditional equilibrium in the lower stretches of the river Po. These results are the first of this kind reported for the whole river course and highlight the factors and mechanisms controlling the origin, mobility and fate of nutrients and trace metals in the river Po. PMID- 12767289 TI - Simultaneous effects of environmental factors on motile Aeromonas dynamics in an urban effluent and in the natural seawater. AB - Seasonal dynamics of motile Aeromonas in a treated urban effluent and in natural seawater along the Sfax coast (Mediterranean sea, Tunisia) were measured over a year concurrently with seven environmental factors, and compared with those of faecal coliforms. Counts for Aeromonas from a standard plate count method, ranged from 1.48 x 10(5)CFU.100 ml(-1) to 2.2 x 10(8)CFU.100 ml(-1) in the effluent and from 7.9 x 10(3)CFU.100 ml(-1) to undetectable level in the surface marine waters. Contrary to faecal coliforms, the Aeromonas dynamics exhibited a seasonal distribution in seawater which was inverse of the seasonal distribution in the sewage: From the end of November 1998 to April 1999 (cold period), Aeromonas counts increased in the treated effluent, while it decreased very rapidly in seawater. From May to October (warm period), Aeromonas abundance decreased in the effluent but showed an increasing fluctuating trend in the marine waters with a maximum in late summer/early autumn when the temperatures were around 22-23 degrees C. Multiple correlation and regression analyses suggest, by the coefficient of determination (R(2)), that 42% of variance in Aeromonas number changes in the treated effluent, may be explained by only turbidity, radiation and Aeromonas density in the previous sample, while 37% of variance in marine ecosystem were explained by radiance and conductivity. Furthermore, the t statistics and their p values and the coefficient of partial determination (r(2)) indicated that radiance contributed the most (r(2)=0.3184, t=-3.2, p=0.0041) to the dynamics of motile Aeromonas in seawater, when combined with conductivity. The models relevant for changes in faecal coliforms abundance incorporated turbidity, radiance in the effluent and conductivity, pH, radiance, turbidity in coastal marine environment. These models explain 66% and 73% of the observed cell number fluctuation, with turbidity (r(2)=0.529, t=5.08, p=0.0001) and conductivity (r(2)=0.5407, t=4.97, p=0.0001) as dominant factors in the multivariate model proposed, respectively, for the two sampling sites. The results presented here suggest that the combination of negative effects of sunlight and conductivity in natural seawater mainly affects the colony-forming capacity and make the motile Aeromonas nonrecoverable during cold months. PMID- 12767290 TI - Kinetics of sorptive removal of chromium(VI) from aqueous solutions by calcined Mg-Al-CO(3) hydrotalcite. AB - The sorptive removal of hexavalent chromium from aqueous solutions by calcined Mg Al-CO(3) hydrotalcite was investigated in a batch mode. The influence of agitation speed, solution pH, initial chromium concentration, sorbent concentration and temperature has been tested in kinetic runs. Three kinetic models have been evaluated to fit the experimental data: the pseudo-first order, the modified-second order and the Elovich equation. It was shown that the first order model could best describe the sorption kinetics. A Freundlich isotherm was used to fit equilibrium experiments. Hydrotalcite presents a sorption capacity of approximately 120 mg Cr/g, under the investigated experimental conditions. The calculated activation energy for process studied was around 40 kJ/mol. PMID- 12767291 TI - Rate constants of reactions of bromine with phenols in aqueous solution. AB - The kinetics of bromination of six ortho- and para-substituted phenols was investigated between pH 5 and pH 12 in aqueous solution. Kinetics was followed with a continuous-flow reactor previously validated by studying the fast reaction between chlorine and ammonia. The overall reaction rate between bromine and phenols is controlled by the reaction of HOBr with the phenoxide ion between pH 6 and pH 10. The reaction of HOBr with the undissociated phenols and the reaction of BrO(-) with the phenoxide ions become only significant for pH<6 and pH>10, respectively. The second-order rate constants for the reaction of HOBr with phenoxide ions vary between 1.4(+/-0.1)x10(3) and 2.1(+/-0.5)x10(8)M(-1)s(-1) for 2,4,6-trichlorophenol and 4-methylphenol, respectively. Hammett-type correlation was obtained for the reaction of HOBr with the phenoxide ions (log(k)=8.0-3.33 x Sigmasigma) and was compared with Hammett-type correlations of HOCl and HOI. The reaction rate of bromine with phenol-like organic compounds was estimated to be about 10(3)-fold higher than with chlorine and 10(3)-fold lower than with ozone in drinking water treatment conditions. PMID- 12767292 TI - A simplified method to assess structurally identifiable parameters in Monod-based activated sludge models. AB - The first step in the estimation of parameters of models applied for data interpretation should always be an investigation of the identifiability of the model parameters. In this study the structural identifiability of the model parameters of Monod-based activated sludge models (ASM) was studied. In an illustrative example it was assumed that respirometric (dissolved oxygen or oxygen uptake rates) and titrimetric (cumulative proton production) measurements were available for the characterisation of nitrification. Two model structures, including the presence and absence of significant growth for description of long- and short-term experiments, respectively, were considered. The structural identifiability was studied via the series expansion methods. It was proven that the autotrophic yield becomes uniquely identifiable when combined respirometric and titrimetric data are assumed for the characterisation of nitrification. The most remarkable result of the study was, however, that the identifiability results could be generalised by applying a set of ASM1 matrix based generalisation rules. It appeared that the identifiable parameter combinations could be predicted directly based on the knowledge of the process model under study (in ASM1-like matrix representation), the measured variables and the biodegradable substrate considered. This generalisation reduces the time consuming task of deriving the structurally identifiable model parameters significantly and helps the user to obtain these directly without the necessity to go too deeply into the mathematical background of structural identifiability. PMID- 12767293 TI - Evaluation of polyethylene hollow-fiber membranes for hydrogen delivery to support reductive dechlorination in a soil column. AB - Engineered systems are often needed to supply an electron donor, such as hydrogen (H(2)), to the subsurface to stimulate the biological dehalogenation of perchloroethene (PCE) to ethene. A column study was performed to evaluate the ability of gas permeable hollow-fiber membranes to supply H(2) directly to PCE contaminated groundwater to facilitate bioremediation. Two glass columns were packed with soil obtained from a trichloroethene-contaminated site at Cape Canaveral, Florida, and were fed a minimal medium spiked with PCE (7 microM) for 391 days. The columns were operated in parallel, with one column receiving H(2) via polyethylene hollow-fiber membranes (lumen H(2) pressure of approximately 1atm) and a control column receiving no H(2). PCE was initially dechlorinated at a similar rate and to a similar extent in both columns, likely due to the presence of soil organic matter that was able to support dechlorination. After 265 days of operation, dechlorination performance declined in the control column and the benefits of membrane-supplied H(2) became evident. Although the membrane supplied H(2) effectively stimulated PCE dechlorination at the end of the experiment (days 359-391), the system was inefficient in that only 5% of the supplied H(2) was used for dechlorination. Most of the remainder was used to support methanogenesis (94%). Despite the dominance of methanogens, nearly complete dechlorination of PCE to ethene was observed in the H(2)-fed column. In addition to the inefficient use of H(2), operational problems included excessive foulant accumulation on the outside of the membrane fibers and water condensation inside the fibers. Use of alternative membrane materials and changes to the operating approach (e.g. pulsing or supplying H(2) at low partial pressures) may help to overcome these problems so that this technology can provide effective and stable remediation of aquifers contaminated with chlorinated ethenes. PMID- 12767294 TI - Application of gas-inducing reactor to obtain high oxygen dissolution in aeration process. AB - The application of gas-inducing reactor to obtain high oxygen dissolution has been investigated at various operation conditions including agitation speed, temperature (20-40 degrees C), pressure (1.0-1.2 atm) and working liquid levels. Correlations regarding onset speed, agitation power consumption, gas holdup and oxygen mass transfer coefficient were established from experimental data. Onset speed can be accurately predicted with modified Froude number. The agitation power consumptions before and after onset speed is a function of Froude number and working liquid level. Gas holdup is an important factor influencing the mass transfer of oxygen after onset speed. In the study of mass transfer of oxygen (T=20 degrees C, P=1 atm), the highest dissolved oxygen concentration is as high as 39.34-39.92 mg x L(-1). The value of k(L)a is within 0.511-1.792 min(-1). The k(L)a is not affected by the oxygen gas pressure (1.0-1.2 atm). However, k(L)a increases with increasing temperature. The gas-inducing reactor of this study has higher k(L)a than the other type of gas-inducing reactor under the same unit volume power consumption. The pure oxygen utilization rate of this system can be as high as 100%. PMID- 12767295 TI - Bacterial immobilization and oxidation of arsenic in acid mine drainage (Carnoules creek, France). AB - The acid waters (pH=2.73-3.37) originating from the Carnoules mine tailings contain high dissolved concentrations of arsenic (1-3.5 mmol l(-1)) and iron (20 40 mmol l(-1)). At the outlet, arsenite predominates. During the first 30 m of downflow, 20-60% is removed by coprecipitation with Fe(III). This process results from bacterially mediated As- and Fe-oxidation. The precipitation rates in the creek depend on the oxygen concentration in spring water and are lower during the dry summer period when the anoxic character of the spring water inhibits the activity of oxidizing bacteria. Ex situ experiments show that the presence of bacteria-rich precipitates increases the As- and Fe-removal rates. Three strains of bacteria promoting the oxidation of As have been isolated, and two of them have the characteristics of Thiomonas ynys1. The third strain, which is not identified yet, also catalyzes the oxidation of Fe. PMID- 12767296 TI - Effects of substrate concentrations on the growth of heterotrophic bacteria and algae in secondary facultative ponds. AB - This paper presents the effect of substrate concentration on the growth of a mixed culture of algae and heterotrophic bacteria in secondary facultative ponds (SFPs) utilizing settled domestic sewage as a sole source of organic carbon. The growth of the mixed culture was studied at the concentrations ranging between 200 and 800 mg COD/l in a series of batch chemostat reactors. From the laboratory data, the specific growth rate (micro) was determined using the modified Gompertz model. The maximum specific growth rate ( micro(max)) and half saturation coefficients (K(s)) were calculated using the Monod kinetic equation. The maximum observed growth rate ( micro(max)) for heterotrophic bacteria was 3.8 day(-1) with K(s) of 200 mg COD/l. The micro(max) for algal biomass based on suspended volatile solids was 2.7 day(-1) with K(s) of 110 mg COD/l. The micro(max) of algae based on the chlorophyll-a was 3.5 day(-1) at K(s) of 50mg COD/l. The observed specific substrate removal by heterotrophic bacteria varied between the concentrations of substrate used and the average value was 0.82 (mg COD/mg biomass). The specific substrate utilization rate in the bioreactors was direct proportional to the specific growth rate. Hence, the determined Monod kinetic parameters are useful for the definition of the operation of SFPs. PMID- 12767297 TI - The characteristics of the bacterial community structure and population dynamics for phosphorus removal in SBR activated sludge processes. AB - The characteristics of the bacterial community structure and population dynamics for phosphorus removal in activated sludge were investigated using laboratory scale sequencing batch reactor (SBR) activated sludge processes fed with four different carbon sources. Phosphorus removal activity, quinone profile of the activated sludge and isolated bacterial strains were monitored. An enhanced phosphorus removal activity was accompanied by the increase in ubiquinone-9 (Q 9). The relationships between phosphorus removal activity and Q-9 contents of the isolated bacterial strains were dependent neither on the organic substrates nor on the species of isolated bacterial strains. The enhanced phosphorus removal capability of activated sludge seemed to be due to the increase in the populations of bacteria with activity of phosphorus removal, i.e. species succession of bacteria in activated sludge ecosystems, rather than the increase in the phosphorus removal capability of phosphorus removal bacteria. Not only Acinetobacter but also Pseudomonas identified from isolated strains were regarded as representative polyphosphates-accumulating organisms and predominant species to the whole of bacterial population in activated sludge for phosphorus removal. PMID- 12767298 TI - Continuous biotransformation and removal of nitrophenols under denitrifying conditions. AB - The effect of COD/NO(3)(-)-N ratio on the biotransformation and removal of 2 nitrophenol (2-NP), 4-nitrophenol (4-NP), and 2,4-dinitrophenol (2,4-DNP) was studied in bench scale upflow anaerobic sludge blanket (UASB) reactors. Sodium acetate and sodium nitrate were used as electron donor (substrate) and electron acceptor, respectively. Nitrate nitrogen loading was increased from 0.098 to 0.6 kg/m(3)d in order to keep COD/NO(3)(-)-N ratio as 20.8, 14.3, 9.8, 5.0, 4.0 and 3.33. Throughout the study, input nitrophenolic concentration and hydraulic retention time (HRT) were kept constant as 30 mg/l and 24h, respectively. 2 Aminophenol (2-AP), 4-aminophenol (4-AP) and 2-amino,4-nitrophenol (2-A,4-NP) were found as the major intermediate metabolite of 2-NP, 4-NP and 2,4-DNP, respectively. Removal of all the three nitrophenols increased with lowering of COD/NO(3)(-)-N ratio. However, nitrophenols removal got adversely affected when COD/NO(3)(-)-N ratio was reduced below 5. Maximum removal achieved were 91.63%, 90.17% and 86.10% for 2-NP, 4-NP and 2,4-DNP, respectively at a COD/NO(3)(-)-N ratio of 5. Simultaneous denitrification and methanogenesis was observed in all the reactors throughout the study. PMID- 12767299 TI - Short- and long-term sorption/desorption of polycyclic aromatic hydrocarbons onto artificial solids: effects of particle and pore sizes and organic matters. AB - In order to elucidate the effect of nanopores and organic matters on sequestration of contaminants, short- and long-term sorption and desorption of naphthalene and pyrene in seven artificial solid-water systems were studied. Fast sorption occurred in every case; steady states were reached within 1-5h. Sorption constants varied drastically among the seven absorbents, ranging from 1.19 to 5.29 x 10(3) for naphthalene, and from 24.3 to 6.52 x 10(4) for pyrene. Slow sorption continued to take place in some cases, especially on absorbents containing humic matter. Desorption usually took place in two stages, fast and slow, on both unaged and aged absorbents. Irreversibility of desorption occurred for every absorbent except for silica particles modified with octadecyl silyl. Aging led to a reduction of fast desorption fraction due to entrapment of the chemicals into nanopores and partitioning of the chemicals into condensed areas of humic matters, and showed no effect on slow desorption and irreversibility of desorption. On the whole, entrapment into nanopores and partitioning into humic matters are considered to be important for sequestration of contaminants. Irreversibility of desorption is considered to be a more influencing factor than percentage of desorption for describing the extent of sequestration. PMID- 12767300 TI - Long-term trends in water quality and their impact on macroinvertebrate assemblages in eutrophic lowland rivers. AB - Long-term trends in water quality in eutrophic lowland rivers in eastern England were investigated and their impact on macroinvertebrate assemblages studied. Dissolved oxygen (DO) declined significantly in eight rivers in Essex and Suffolk over 40 years to 1998. Chloride concentrations significantly increased during the same period in most rivers. Total oxidized nitrogen and soluble reactive phosphorus increased until the 1980s, then began to decline. Biotic scores (Lincoln Quality Index) generally increased over 14 years to 1998 and there were significant positive relationships between biotic scores and several nutrients. Invertebrate families and environmental variables sampled over the eight rivers in a dry year (1997) and a wet year (1998) were subjected to multivariate analysis. River stretches were grouped according to substrate requirements of indicator invertebrates. In the dry year, those river stretches behind mills or immediately downstream of sewage treatment works (STW) were grouped. In the wet year, there was only one separate group, comprising sites downstream of STWs. Nutrients, DO and low flows have a much greater influence on water quality, and hence invertebrate assemblages, during drought years than during wet years. PMID- 12767301 TI - Effect of initial microbial density on inactivation of Giardia muris by ozone. AB - Inactivation of microorganisms by disinfectants frequently shows non-linear behavior on a semilogarithmic plot of log survival ratio versus time. A number of models have been developed to depict these deviations from Chick's Law. Some of the models predict that the log survival ratio (at a particular disinfectant dose and contact time, even in absence of demand) would be a function of the initial concentration of microorganisms (N(0)), while other models do not predict such an effect. The effect of N(0) on the survival ratio has not been deliberately tested. This work examined the inactivation of Giardia muris by ozone in batch systems, deliberately varying the disinfectant dose and N(0). It was found that the models predicting a dependency of survival on N(0) gave a better description to the data than models that did not predict such a dependency. Hence there is an apparent decrease in disinfection efficiency of ozone against Giardia muris (at pH 8 and 15 degrees C) as the initial microorganism concentration decreases. This phenomena should be taken into account by both disinfection researchers and by process design engineers. PMID- 12767302 TI - Fouling characteristics of NF and RO operated for removal of dissolved matter from groundwater. AB - A pilot study had been performed for about 6 months in order to investigate the removal efficiency of dissolved matter and its fouling potential during nanofiltration (NF) and reverse osmosis (RO) of local groundwater that was pretreated with an ultrafiltration (UF) membrane system. After pilot plant operation, autopsy tests were performed to identify the characteristics of foulants that were attached to the membrane surface. In the autopsy tests, the flux recovery for each specific cleaning scheme (hydraulic washing, acid cleaning, and alkaline cleaning) was also measured using a dead-end filtration cell unit. The washing solution used in each chemical cleaning was analyzed to identify major components of the foulants, and the membrane surface was observed using the scanning electron microscopy (SEM). Among three kinds of membranes tested, one NF and two RO membranes, the NF and RO1 membranes showed a rapid flux decline after 100 days of operation. Especially, the RO1 membrane showed the more serious flux decline than the NF membrane. The RO2 membrane, with the lowest recovery rate, demonstrated a gradual flux decline. The removal efficiency of dissolved inorganic matter (as conductivity) for each NF, RO1 and RO2 membrane was 76.3%, 88.2% and 95.3%, respectively. The removal of dissolved organic matter (as total organic carbon) was found to be about 80% for both NF and RO membranes used in this study. During the membrane autopsy tests, five sections of the fouled membrane were cut along each NF and RO membrane module from the feed inlet side to the concentrate outlet side, the specific flux for each membrane section was measured before and after each cleaning step. As expected, the degree of fouling was intensified along the membrane surface as the feed flow approached the outlet. Based on the analysis results of wash water used in each cleaning step, the major foulants attached to the membrane surface appeared to be Ca bound with inorganic matter and Si bound with organic matter. Fe seemed to be a great contributor to irreversible fouling. The SEM analysis indicated that the organic matter was forming the first fouling layer close to the membrane and that the inorganic matter was layered top of the organic fouling layer in a tetragonal shape. Any evidence of biofouling was not observed in this study because most of microorganisms had been already removed by the UF pretreatment. PMID- 12767303 TI - Removal of viable and inactivated Cryptosporidium by dual- and tri-media filtration. AB - The limited efficacy of disinfectants, other than ultraviolet irradiation and ozonation, as a barrier against Cryptosporidium parvum in drinking water treatment has underscored the increased importance of oocyst removal by filtration. Currently, no reliable surrogates have been identified for C. parvum removal by filtration. As a result, evaluations of the Cryptosporidium removal by treatment operations have been performed using oocysts. It has typically been assumed that chemically inactivated oocysts are suitable surrogates for viable oocysts. Measurements of electrophoretic mobility, however, have shown that chemical inactivation changes the surface charge of Cryptosporidium oocysts. The present bench-scale research indicated that formalin-inactivated oocysts are reliable surrogates for viable oocysts during both stable filter operation and periods where filtration processes are challenged, such as coagulation failure. This finding is important because of the practical difficulties associated with using viable oocysts in filtration investigations. Poor coagulation conditions severely compromised removal of viable and inactivated oocysts by dual- and tri media filters compared to stable operating conditions and filter ripening, emphasizing the importance of optimized chemical pre-treatment (coagulation) for the successful removal of oocysts during filtration. The treatment optimization experiments also indicated that tri-media filters offered only marginally higher oocyst removals than dual-media filters. PMID- 12767304 TI - The effect of residual ammonia concentration under aerobic conditions on the growth of Microthrix parvicella in biological nutrient removal plants. AB - It is demonstrated with two parallel single reactor intermittently aerated nitrification denitrification systems fed municipal wastewater as influent, that Microthrix parvicella bulking can be stimulated and cured by manipulating the ammonia concentration in the aerobic period (by inhibiting the nitrifiers) to high and low values respectively. The proliferation or not of M. parvicella is hypothesized to be due to their requirement for ammonia as a nitrogen source for growth. In terms of this hypothesis, if nitrification is rapid and complete, ammonia is not freely available and will limit M. parvicella growth. If nitrification is not complete for whatever reason, ammonia is available for the growth of the slow growing M. parvicella, enabling their proliferation to cause a bulking sludge. This hypothesis does not overturn or replace the anoxic-aerobic (AA, or low Food/Microorganism, F/M, ratio) filament bulking hypothesis of Casey et al. (Water SA 25(4) (1999) 425) but appears to be additional to it. Future research will focus on determining how elements of both hypotheses superimpose on the conditions in BNR systems, to produce an AA filament bulking sludge or not. PMID- 12767306 TI - Kinetic analysis for the removal of a reactive dye from aqueous solution onto hydrotalcite by adsorption. AB - The removal of a reactive color, Cibacron Yellow LS-R, from aqueous solutions by adsorption onto hydrotalcite particles is investigated using batch rate experiments. Measurements are performed at various initial color concentrations, solid loads, pH values and ionic backgrounds (dissolved NaCl). The speed of agitation and the temperature inside the batch adsorber are also varied within a practical range of values. It is shown that the sorption capacity is relatively high for most experimental conditions so hydrotalcite may be considered as a suitable sorbent for this application. The probable mechanism of the process is investigated by a number of homogeneous and heterogeneous reaction kinetic models as well as diffusion kinetic models. It is found that no single kinetic model can fully describe the sorption process at all times. At least three independent rate controlling mechanisms appear to compete each other and dominate the different stages of sorption. PMID- 12767305 TI - Remediation of alpha-methylnaphthalene-contaminated sediments using peroxy acid. AB - Laboratory experiments have been conducted to elucidate the degradation of alpha methylnaphthalene in lake sediments using an advanced oxidation process (AOP) employing peroxy-acids as oxidizing agents. Abiotic degradation of alpha methylnaphthalene was observed in sediment samples exposed to a 1:1:1 v/v/v mixture of water/organic acid/hydrogen peroxide solution. Sediment samples were collected from Lake Macatawa (Holland, MI) and ranged in total organic carbon content from 2.1% (sandy sediment sample III) to 12.8% (silty-clay sediment sample I) and surface area, which ranged from 3.2m(2)/g (sandy sediment sample III) to 22.0m(2)/g (silty-clay sediment sample I). Analytical measurements performed by gas chromatography revealed varied rates of degradation, depending on the type of acid and the characteristics of the sediment sample. Within 24h, alpha-methylnaphthalene was reduced to 70% and 100% of its original concentration when propionic acid and acetic acid were used as the organic acids, respectively. The formation of products was explored using a gas chromatograph equipped with a mass spectrometer and indicated mainly the formation of lower molecular weight compounds, such as alkyl chains. This AOP method of PAH degradation in sediment/liquid slurry was fast acting and products were most likely biodegradable. PMID- 12767307 TI - Comparison of nutrient and contaminant fluxes in two areas with different hydrological regimes (Emporda Wetlands, NE Spain). AB - Nutrient (N and P), heavy metal (Ni, Cd, Cr, Cu and Pb) and pesticide (DDT, DDD, DDE, lindane, aldrin, endrin, dieldrin, permethrin, atrazine and simazine) concentrations in water and sediment were analysed in the Emporda Wetlands, a Mediterranean wetland area in NE Spain. Mean nutrient and contaminant concentrations and input and output loads via tributaries were compared in two marshes with different water turnover: a freshwater marsh (FWM), with a high water turnover rate due to continuous surface water inputs and outputs, and a brackish water marsh (BWM), with lower turnover and no continuous surface output, where water remains confined during dry periods. Mean concentrations of most heavy metals exceeded the maximum permissible concentration (MPC) in BWM, whilst only some pesticides reached MPC in FWM. The confined waters of BWM showed higher sensitivity to contaminant input loadings than FWM due to the lack of continuous water outputs. Non-point source pollution inputs during runoff (mainly in FWM) and concentration during confinement (mainly in BWM) showed as the main environmental problems related to nutrients and contaminants in these ecosystems. Thus, the importance of confinement and its effect on pollutant concentrations must be borne in mind to achieve correct management of Mediterranean wetlands. PMID- 12767308 TI - Product formation from thiophene by a mixed bacterial culture. Influence of benzene as growth substrate. AB - The influence of benzene as a growth substrate on the cometabolic conversion of thiophene was investigated in batch systems with microorganisms originating from an creosote contaminated site. Benzene was shown to stimulate the conversion of thiophene with a first-order rate, during the initial phase of transformation. The microorganisms were able to transform thiophene in the absence of benzene at a zero-order rate. Thiophene was converted to five oxidation products, regardless of the presence of benzene. Benzene had no influence on the distribution of these oxidation products. The main oxidation product, a thiophene sulphoxide dimer, represented 78+/-12% of the transformed thiophene, while the second most important product, also a thiophene sulphoxide dimer, represented 20+/-2% of the converted thiophene. PMID- 12767310 TI - Pupil responses associated with coloured afterimages are mediated by the magno cellular pathway. AB - Sustained fixation of a bright coloured stimulus will, on extinction of the stimulus and continued steady fixation, induce an afterimage whose colour is complementary to that of the initial stimulus; an effect thought to be caused by fatigue of cones and/or of cone-opponent processes to different colours. However, to date, very little is known about the specific pathway that causes the coloured afterimage. Using isoluminant coloured stimuli recent studies have shown that pupil constriction is induced by onset and offset of the stimulus, the latter being attributed specifically to the subsequent emergence of the coloured afterimage. The aim of the study was to investigate how the offset pupillary constriction is generated in terms of input signals from discrete functional elements of the magno- and/or parvo-cellular pathways, which are known principally to convey, respectively, luminance and colour signals. Changes in pupil size were monitored continuously by digital analysis of an infra-red image of the pupil while observers viewed isoluminant green pulsed, ramped or luminance masked stimuli presented on a computer monitor. It was found that the amplitude of the offset pupillary constriction decreases when a pulsed stimulus is replaced by a temporally ramped stimulus and is eliminated by a luminance mask. These findings indicate for the first time that pupillary constriction associated with a coloured afterimage is mediated by the magno-cellular pathway. PMID- 12767309 TI - Chromatic induction and the layout of colours within a complex scene. AB - A target's apparent colour is influenced by the colours in its surrounding. If the surrounding consists of a single coloured surface, the influence is a shift 'away' from the surface's colour. If the surface is more than 1 degrees from the target area the shift is very small. If there are many surfaces, then not only the average luminance and chromaticity of the surfaces matters, but also the chromatic variability. It is not yet clear whether it makes any difference where the chromatic variability is within the scene, so we constructed stimuli in which the chromatic variability was restricted to certain regions. We found that it made very little difference where the chromatic variability was located. The extent to which the average colour of nearby surfaces influences the apparent colour of the target seems to depend on the average chromatic variability of the whole scene. PMID- 12767311 TI - Dual nonlinearities regulate contrast sensitivity in pattern discrimination tasks. AB - Many current psychophysical models propose that visual processing in cortex is hierarchical, with nonlinearities sandwiched between linear stages of processing. In earlier publications, we proposed a model of this type to account for masking effects found with spatial frequency and orientation discriminations. Our model includes two nonlinear mechanisms that regulate contrast sensitivity in early cortical mechanisms. The first is a local within-pathway nonlinearity that accelerates at low contrasts but is compressive at high. The second is a pooled nonlinear gain control process that operates over a broad range of neurons with different tuning characteristics. Here, we test predictions of the model for spatial frequency discriminations. The model predicts that at low contrasts, adding a grating mask oriented parallel to test gratings will improve discrimination performance via operation of the within-pathway nonlinearity, analogous to the "dipper effect" found with contrast discriminations. Adding an orthogonally oriented mask is predicted to have no effect at low contrasts, where pooled gain control processes contribute little to performance. At high contrasts, the model predicts that performance will asymptote and become independent of contrast with either parallel or orthogonal masks. The results confirm model predictions. PMID- 12767312 TI - The overlay interference task and object-selective visual attention. AB - A series of experiments was carried out to examine object-based visual attention in an interference task. Observers were presented with two transparently overlapping equilateral triangles forming a "Star of David". One of these triangles was darker than the background, the other was lighter than the background. The observers were required to make a speeded choice response to the orientation of the darker triangle. The presence of the light triangle produced a robust interference effect that manifested as a slower response time. This effect was strongly modulated by the relative contrast of the target and distractor triangles. It was reduced when the light distractor triangle was separated in depth from the target triangle. Since the configuration rules out the possibility of 2-D spatial selection, it is concluded that object-based selection occurs in interference tasks and that the effectiveness of this selection is modulated by visual attributes that are not directly relevant to the task. PMID- 12767313 TI - Visual search is facilitated by scene and sequence familiarity in rhesus monkeys. AB - Whether searching for targets in a familiar scene leads to improved performance was tested in monkeys. We found that search performance improved for a familiar scene when target locations were always randomized. However, when target locations repeatedly followed a predictable sequence, performance improvement for a familiar scene was manifested only for targets presented in a familiar sequence, suggesting that scene memory might be masked by the learning of target sequences. These results suggest that information about a visual scene can facilitate the performance of visual search, and that this memory is coupled to the learned sequence of target locations. PMID- 12767314 TI - Effect of sleep deprivation and driving duration on the useful visual field in younger and older subjects during simulator driving. AB - Nine older subjects (40-51 years) and 10 younger subjects (18-30 years) took part in two one-hour driving sessions. They performed a very monotonous task during which they had to follow a vehicle either after a complete night of sleep or after one night of sleep deprivation. While driving their useful visual field was assessed by introducing signals that would appear on the whole road scene. The analysis of the data indicates that the ability to process peripheral signals deteriorates with age, driving duration and sleep deprivation. However, the effects of these three variables on the peripheral visual ability are not similar in a dual task. The driver's useful visual field changes with age and prolongation of the monotonous driving activity according to a tunnel vision phenomenon. On the other hand, a sleep debt deteriorates the useful visual field according to a general interference phenomenon. These results are discussed in terms of decrease in the level of arousal and increase of fatigue. PMID- 12767315 TI - Centric-minded templates for self-motion perception. AB - We propose a two-layer neuromorphic architecture by which motion field pattern, generated during locomotion, are processed by template detectors specialized for gaze-directed self-motion (expansion and rotation). The templates provide a gaze centered computation for analyzing motion field in terms of how it is related to the fixation point (i.e., the fovea). The analysis is performed by relating the vectorial components of the act of motion to variations (i.e., asymmetries) of the local structure of the motion field. Notwithstanding their limited extension in space, such centric-minded templates extract, as a whole, global information from the input flow field, being sensitive to different local instances of the same global property of the vector field with respect to the fixation point; a quantitative analysis, in terms of vectorial operators, evidences this property as tuning curves for heading direction. Model performances, evaluated in several situations characterized by conditions of absence and presence of pursuit eye movements, validate the approach. We observe that the gaze-centered model provides an explicit testable hypothesis that can guide further explorations of visual motion processing in extrastriate cortical areas. PMID- 12767316 TI - Hyperglycemia affects flicker-induced vasodilation in the retina of healthy subjects. AB - Flickering light stimulation of retinal photoreceptors induces retinal vessel dilation in humans. In the present study the effect of high blood glucose levels on this neuro-vascular mechanism was investigated in 12 healthy young male subjects. Blood glucose levels were consecutively increased during 30 min to 100, 200 and 300 mg/dl and kept at the respective level for the following 30 min using hyperglycemic insulin clamps. Eight Hertz flickering light was applied to the fundus at the end of each glucose plateau during continuous retinal vessel diameter measurements with the Zeiss retinal vessel analyser (RVA). During normoglycemia (100 mg/dl) flickering light induced a significant vasodilation of retinal arteries (+2.8+/-0.4%, p<0.0001) and veins (+2.6+/-0.4%, p<0.0001). At 300 mg/dl blood glucose the flicker response in retinal veins was significantly decreased by 55% (p=0.015 versus 100 mg/dl). The modified RVA employed in the present study provides high sensitivity and is capable of studying flicker induced retinal vasodilation. Using this technique the present study confirms that flickering light stimulation of the human retina induces vasodilation in retinal vessels in healthy subjects. In addition, our data indicate that the retinal vessel response to flickering light stimulation is significantly reduced during hyperglycemia in humans. The relevance of this finding for diabetes related eye disease remains to be shown. PMID- 12767317 TI - Continuous annular chromatography. AB - The principle of continuous annular chromatography (CAC) has been known for several decades. CAC is a continuous chromatographic mode, which lends itself to the separation of multi-component mixtures as well as of bi-component ones. In CAC, the mobile and stationary phases move in a crosscurrent fashion, which allows transformation of the typical one-dimensional batch column separation into a continuous two-dimensional one. With the exception of linear gradient elution, all chromatographic modes have at present been applied in CAC. This review focuses on the capacity of CAC for preparative bioseparation. The historical developments and the predecessors of modern CAC are briefly summarized. The state of-the-art in the theoretical prediction and simulation of CAC separations is discussed, followed by an overview of current CAC instrumentation and example applications, especially for the isolation of proteins and other bio(macro)molecules. In this context, issues of scale up as well as method development and transfer from batch to continuous CAC columns are discussed using recent bioseparation efforts as pertinent examples. PMID- 12767318 TI - Large-scale ion-exchange column chromatography of proteins. Comparison of different formats. AB - This review describes the performance of various column designs available to process-scale users of low-pressure chromatography for protein purification. By carrying out a range of ion-exchange separations using Whatman microgranular ion exchange celluloses we are able to compare and contrast the practical performance issues associated with several designs of axial and radial flow columns. PMID- 12767319 TI - Comparison of protein A affinity sorbents. AB - Protein A is a popular generic ligand for purification of monoclonal and recombinant antibodies. The performance of 15 commercially available protein A media was studied. Equilibrium and dynamic binding capacity for human IgG was determined and the capture of IgG from a crude feed-stock was investigated. For initial screening the dynamic binding capacity was determined at small scale. Media with good performance were further tested with increased column height. Comparing the data from the two different column heights it could be shown that the dynamic capacity strongly depends on the residence time. Agarose based media exhibited higher binding capacity at residence times longer than 3 min whereas polymeric media or media based on porous glass showed a lesser dependence on the flow velocity and the residence time. A quantitative description of this behavior was derived by determination of the adsorption isotherms and fitting the breakthrough profiles with the Thomas solution. Agarose based media exhibited higher maximum equilibrium binding capacities and the dissociation constants derived from adsorption isotherms were smaller. The other media exhibited higher apparent rate constants, indicating a faster mass transfer. This can be explained by the smaller particle diameter of these media and it can be assumed that constant pattern conditions are thereby obtained more quickly. Selectivity was tested by performing antibody purification under standardized conditions. Polyclonal human IgG in cell culture supernatant containing 2.5% fetal calf serum was used as a representative feed-stock. Under the applied conditions several sorbents showed very tight binding of IgG and in some cases most of the sample remained on the sorbent. The study can be useful as a guide for optimization of large-scale purification processes. PMID- 12767320 TI - Isolation of antigens and antibodies by affinity chromatography. AB - Antibody-antigen binding constants are commonly strong enough for an effective affinity purification of antibodies (by immobilized antigens) or antigens (by immobilized antibodies) to work out a straightforward purification method. A drawback is that antibodies are large protein molecules and subject to denaturation under conditions required for the elution from the complex. Structures of antigens can vary but usually antigens are also equally subject to similar problems. The lability of the components can sometimes make the procedure sophisticated, but usually in all cases it is possible to find a satisfactory approach. In certain cases, specific interactions of the Fc part of antibodies are more facile to exploit for their purification. PMID- 12767321 TI - Design and selection of ligands for affinity chromatography. AB - Affinity chromatography is potentially the most selective method for protein purification. The technique has the purification power to eliminate steps, increase yields and thereby improve process economics. However, it suffers from problems regarding ligand stability and cost. Some of the most recent advances in this area have explored the power of rational and combinatorial approaches for designing highly selective and stable synthetic affinity ligands. Rational molecular design techniques, which are based on the ability to combine knowledge of protein structures with defined chemical synthesis and advanced computational tools, have made rational ligand design feasible and faster. Combinatorial approaches based on peptide and nucleic acid libraries have permitted the rapid synthesis of new synthetic affinity ligands of potential use in affinity chromatography. The versatility of these approaches suggests that, in the near future, they will become the dominant methods for designing and selection of novel affinity ligands with scale-up potential. PMID- 12767322 TI - Protein purification by affinity precipitation. AB - Developing the most efficient strategy for the purification of a (recombinant) protein especially at large scale remains a challenge. A typical problem of the downstream process of mammalian cell products is, for instance, the early capture of the highly diluted product from the complex process stream. Affinity precipitation has been suggested in this context. The technique is known for over 20 years, but has recently received more attention due to the development of new materials for its implementation, but also because it seems ideally suited to specific product capture at large scale. The present review gives a comprehensive overview over this technique. Besides an introduction to the basic principle and a brief summary of the historical development, the main focus is on the current state-of-art of the technique, the available materials, important recent applications, as well as process design strategies and operating procedures. Special consideration is given to affinity precipitation for product recovery at large scale. PMID- 12767323 TI - Role of mass spectrometry in the purification of peptides and proteins. AB - Experiments were carried out to evaluate the fractionation of proteins and peptides according to mass. Model mixtures were separated by either reversed phase or ion-exchange chromatography with mass spectrometry-compatible mobile phase additives. Fraction collection was triggered by the mass/charge ratio of each one of the components of the mixture. Chromatography was additionally monitored with a UV-Vis detector in order to compare the new technique with generally accepted in separations. The results indicated that adequate purification is achieved by this new technique. Fraction collection triggered by changes in the mass/charge ratio reduces sample handling and analysis time. This study demonstrates the utility of mass-directed fractionation of peptides and proteins when mass spectrometry-compatible mobile phase additives are used. PMID- 12767324 TI - Hydrophobic interaction chromatography of proteins. II. Binding capacity, recovery and mass transfer properties. AB - Hydrophobic interaction chromatography media suited for large scale separations were compared regarding dynamic binding capacity, recovery and mass transfer properties. In all cases, pore diffusion was the rate limiting step. Reduced heights equivalent to a theoretical plate for bovine serum albumin derived from breakthrough curves at reduced velocities between 60 and 1500 ranged from 10 to 700. Pore diffusion coefficients were derived from pulse response experiments for the model proteins alpha-lactalbumin, lysozyme, beta-lactoglobulin, bovine serum albumin and immunoglobulin G. Diffusivity of lysozyme did not follow the trend of decreasing diffusivity with increasing molecular mass, as observed for the rest of the proteins. In general, mass transfer coefficients were smaller compared to ion-exchange chromatography. Dynamic binding capacities for the model protein bovine serum albumin varied within a broad range. However, sorbents based on polymethacrylate showed a lower dynamic capacity than media based on Sepharose. Some sorbents could be clustered regarding binding capacity affected by salt. These sorbents exhibited a disproportional increase of binding capacity with increasing ammonium sulfate concentration. Recovery of proteins above 75% could be observed for all sorbents. Several sorbents showed a recovery close to 100%. PMID- 12767325 TI - Direct observation of intraparticle equilibration and the rate-limiting step in adsorption of proteins in chromatographic adsorbents with confocal laser scanning microscopy. AB - The adsorption of different proteins in a single biospecific and hydrophobic adsorbent particle for preparative protein chromatography has been observed directly by confocal laser scanning microscopy as a function of time at a constant bulk concentration c(b). The bulk concentration was in the non-linear part of the adsorption isotherm. At all times the concentration of free protein at the particle surface was almost equal to the bulk content indicating that external mass transfer resistance is not rate limiting for the adsorption under these conditions. Inside the particles a distinct maximum in adsorbed and free protein concentration that moved inside to a distance of approximately 0.2 R (R particle radius) from the particle surface, was observed. This is due to a decreasing solid-phase density and adsorptive capacity in the particle between 0.8 R and R indicating that the fraction of macropores (or void space) is larger in the outer than in the inner part of the adsorbent particles. By increasing the bulk concentration by a factor of 10 the equilibration time was reduced by about the same magnitude. This is in agreement with the concentration dependence of the effective pore diffusion coefficient D(p,eff)=D(p)/[epsilon(p)[1+nK/(K +c)(2)]] derived from the mass conservation relations describing the adsorption process. The time dependence protein adsorption up to approximately 90% of the equilibration value q* could be described by a bilinear free driving force model. The rapid equilibration in the outer part of the particle with a half-life time of approximately 100 s in the studied systems accounted for 0.3-0.4 q*. The slower equilibration with a up to ten times longer half-life time, was the adsorption in the inner part of the particle that outside 0.5 R accounts for 0.5 0.6 q*. These data were compared with literature data for batch adsorption of proteins in biospecific, hydrophobic and ion-exchange adsorbents. They could also be described by a bilinear free driving force model, with about the same quantitative results as obtained for similar conditions in the single particle experiments. The static adsorption parameters, maximum binding site concentration n, and dissociation constant for the protein binding to a binding site K, were determined from Scatchard plots. For the same protein-adsorbent system the plots changed from linear to non-linear with increasing n. This change occurred when the average distance between adjacent binding sites become of the same order of magnitude as the size of the binding site or adsorbed protein. This causes a shielding of free binding sites increasing with n and the concentration of adsorbed protein, yielding a concentration dependence in K. These results show that for a high throughput and rapid adsorption in preparative chromatography, the adsorption step should be carried out in the non-linear part of the adsorption isotherm with concentrations up to c(b) where q*/c(b)>/=10 to obtain high protein recoveries. To avoid tailing due to the flow of adsorbed proteins in the inner part of the particles further into the particles at the start of the desorption, and to speed up desorption rates, protein adsorption in the particle within 0.5 R from the particle center should be avoided. This requires the further development of suitable pellicular particles for preparative protein chromatography that meet this requirement. PMID- 12767326 TI - Protein binding to polymer brush, based on ion-exchange, hydrophobic, and affinity interactions. AB - The major limitations associated with conventional packed bed chromatography for protein separation and purification can be overcome by using adsorptive microporous membranes as chromatographic media. Microporous membranes have advantages as support matrices in comparison to conventional bead supports because they are not compressible and they eliminate diffusion limitations. As a result, higher throughput and shorter processing times are possible using these membrane systems. In this paper, we review the current state of development in the area of attaching functionalized polymer brushes onto a microporous membrane to form a novel chromatographic medium for protein separation and purification. The functionalized polymer brushes were appended onto the pore surface of a microporous hollow-fiber membrane uniformly across the membrane thickness by radiation-induced graft polymerization and subsequent chemical modifications. We review various applications of this adsorptive membrane chromatography by focusing on polymer brushes bearing ion-exchange, hydrophobic and affinity groups. Proteins were captured in multilayers by the ion-exchange group containing polymer brushes due to the formation of a three-dimensional space for protein binding via the electrostatic repulsion of the polymer brushes. In contrast, proteins were captured in a monolayer at most by the polymer brushes containing hydrophobic or affinity ligands. By permeating a protein solution through the pores rimmed by the polymer brushes, an ideal capturing rate of the proteins with a negligible diffusional mass-transfer resistance was achieved by the functionalized polymer brushes, based on ion-exchange, hydrophobic, and affinity interactions. PMID- 12767327 TI - Characterization and optimization of a chromatographic process based on ethylenediamine-N,N,N',N'-tetra(methylphosphonic) acid-modified zirconia particles. AB - The primary objective of work was to characterize, optimize and model a chromatographic process based on ethylenediamine-N,N,N',N' tetra(methylphosphonic) acid (EDTPA)-modified zirconia particles. Zirconia particles were produced by spray-drying colloidal zirconia. Zirconia spheres produced were further classified, calcined and modified with EDTPA to yield a solid-phase support for use in bio-chromatography (r_PEZ). Specifically, the ability of r_PEZ to selectively bind and enrich IgG, IgA, and IgM from biological fluids was evaluated and demonstrated. To better understand the force of interaction between the IgG and the r_PEZ, the equilibrium disassociation constant (K(d)) was determined by static binding isotherms, as a function of temperature and by frontal analysis at different linear velocities. The maximum static binding capacity (Q(max)) was found to be in the range 55-65 mg IgG per ml of beads, and unaffected by temperature. The maximum dynamic binding capacity (Q(x)) was found to be in the range 20-12 mg IgG per ml of beads. The adsorption rate constant (k(a)) was determined by a split-peak approach to be between 982 and 3242 l mol(-1) s(-1) depending on the linear velocity. The standard enthalpy and entropy values were estimated for this interaction of IgG with this novel support. PMID- 12767328 TI - Evaluation of chromatographic recycling for imidazole used in the chromatographic purification of His-tag recombinant proteins. AB - The aim of this work was to test a recycling method for imidazole used in immobilized metal affinity chromatography (IMAC) as eluent for recombinant histidine-tag (His-tag) protein. After evaluating two supports, the method was optimized with a mixture of bovine serum albumin, sodium chloride and imidazole. Recycling was performed with an eluate fraction from IMAC of His-tag enhanced green fluorescent protein produced in our laboratory and pure imidazole was recovered in water and was analyzed after being freeze-dried. The imidazole was then reused as eluent in IMAC without any modification in its structure or behavior. This procedure can be used for large-scale chromatography. PMID- 12767330 TI - Application of semi-industrial monolithic columns for downstream processing of clotting factor IX. AB - It has been shown in a previous study that monolithic columns can be used for downstream processing of different concentrates of clotting factor IX [K. Branovic et al., J. Chromatogr. A 903 (2000) 21]. This paper demonstrates that such supports are useful tools also at an early stage of the purification process of factor IX from human plasma. Starting with the eluate after solid-phase extraction with DEAE-Sephadex, the use of monolithic columns has allowed much better purification than that achieved with conventional anion-exchange supports. The period of time required for separation is also much reduced. In up-scaling experiments, separations are carried out with 8, 80 and 500 ml columns. A volume of 1830 ml of DEAE-Sephadex eluate, containing a total of 27.6 g of protein and 48500 IU of factor IX is applied to the 500 ml monolithic column. This corresponds to a separation on a pilot scale. The results of this separation after up-scaling are comparable to those obtained with the 8 ml column on a laboratory scale. PMID- 12767331 TI - Preparation of vitamin K-dependent proteins, such as clotting factors II, VII, IX and X and clotting inhibitor protein C. AB - A review is given of preparative methods for the isolation of the vitamin K dependent clotting factors II, VII, IX, X and clotting inhibitor protein C, all derived from human plasma. Factor II, activated factor VII and activated protein C are also obtained from recombinant animal cells. The methods for their purification are described. The problem of difference in posttranslational modifications between plasma derived and recombinant protein is discussed with regard to therapeutic proteins. PMID- 12767332 TI - Chromatographic purification and properties of a therapeutic human protein C concentrate. AB - Protein C deficiency (inherited and acquired) has a relatively high incidence rate in the general population worldwide. For many years, protein C deficient patients have been treated with fresh frozen plasma, prothrombin complex concentrates, heparin or oral anticoagulants, which all have clinical drawbacks. We report the production process of a highly purified human protein C concentrate from 1500 l of cryo-poor plasma by a four-step chromatographic procedure. After DEAE-Sephadex adsorption, protein C was separated from clotting factors II, VII and IX by DEAE-Sepharose FF and further purified, using a new strategy, by an on line chromatographic system combining DMAE-Fractogel and heparin-Sepharose CL-6B. In addition, the product was treated against viral risks by solvent-detergent and nanofiltration on 15-nm membranes. The protein C concentrate was essentially free of other vitamin K-dependent proteins. Proteolytic activity was undetectable. Neither activated protein C, prekallikrein activator, nor activated vitamin K dependent clotting factors were found resulting in good stability of the protein C activity. In vitro and in vivo animal tests did not reveal any sign of potential thrombogenicity. The final freeze-dried product had a mean protein C concentration of 58 IU/ml and a mean specific activity of 215 IU/mg protein, corresponding to over 12000-fold purification from plasma. Therefore, this concentrate appears to be of potential benefit for the treatment of protein C deficiency. PMID- 12767329 TI - Whey proteins as a model system for chromatographic separation of proteins. AB - Although chromatographic separation of whey proteins has been considered too expensive, whey may serve as an excellent model mixture to investigate and validate the use of simulation tools in the development and optimization of chromatographic separations and the outcome could easily be utilized since the model system has an intrinsic value. Besides, milk from transgenic animals could be an attractive source of pharmaceuticals which must be separated from the other proteins in the milk. Several whey proteins are of interest especially, alpha lactalbumin, beta-lactoglobulins, immunoglobulins, lactoperoxidase, and lactoferrin. The scope of the project is to develop a consistent set of chromatographic data for whey proteins including isotherms, transport properties and scale-up studies and to develop the appropriate models for the anion exchangers Q-Sepharose XL, Source 30Q, Ceramic Q-HyperD F, and Merck Fractogel EMD TMAE 650 (S). In this work we have determined and correlated gradient and isocratic retention volumes in the linear range of the isotherm for alpha lactalbumin, beta-lactoglobulin A and B, and bovine serum albumin at a pH from 6 to 9 at various NaCl concentrations. PMID- 12767333 TI - Purification of human plasma haptoglobin by hemoglobin-affinity column chromatography. AB - Haptoglobin (Hp) is an acute-phase protein; its plasma levels increase consistently in response to infection and inflammation. The concentration of human plasma Hp is ranged between 1 and 1.5 mg/ml. Similar to blood type, individual human Hp is classified as Hp 1-1, 2-1, or 2-2. The structural and functional analysis of the Hp, however, has not been studied in detail due to its difficult isolation procedure. Previously, we reported a single step for the purification of porcine Hp. In this study, we established a purification method using a high capacity hemoglobin-affinity column. Briefly, DEAE-purified human hemoglobin was first coupled to Sepharose 4B to prepare an affinity column in a 15-ml bed volume. Following a flow through of human plasma and an extensive wash, the bound material was eluted with a solution of 0.15 M NaCl, pH 11 (adjusted by ammonium), to remove low-affinity bound proteins. The high-affinity bound Hp was then eluted with 0.15 M NaCl containing 5 M urea, pH 11, and collected in tubes containing 100 microl of 1 M Tris buffer, pH 7.0. The biological activity of dialyzed Hp was retained as it formed a complex with hemoglobin on a sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Using this procedure, approximately 10 mg of Hp 1-1, with homogeneity greater than 96%, was obtained from 15 ml of human plasma. Affinity purified Hp 2-1 or 2-2, however, contained trace amounts of apoA-I with the similar approach. The Hp could be further purified by HPLC using a Superose 12 gel-permeation chromatography, if desired, to achieve 100% purity. All the phenotypes of purified Hp consisted of alpha and beta chains on SDS-PAGE in the presence of a reducing reagent, further confirmed by a Western blot analysis. We conclude that human hemoglobin-affinity column was most suitable for the isolation of Hp 1-1 in large quantities. Whereas, one additional step using a gel-permeation was necessary for that of Hp 2-1 and 2-2. PMID- 12767334 TI - Purification of an oxidation-sensitive enzyme, pI258 arsenate reductase from Staphylococcus aureus. AB - Arsenate reductase (ArsC) from Staphylococcus aureus pI258 is extremely sensitive to oxidative inactivation. The presence of oxidized ArsC forms was not that critical for NMR, but kinetics and crystallization required an extra reversed phase purification to increase sample homogeneity. The salt ions observed in the X-ray electron density of ArsC were investigated. Carbonate was found to have the lowest dissociation constant for activation (K(a)=1.1 mM) and potassium was stabilizing ArsC (DeltaT(m)=+6.2 degrees C). Also due to the use of these salt ions, the final yield of the purification had improved with a factor of four, i.e. 73 mg/l culture. PMID- 12767335 TI - Fractionation and purification of the enzymes stored in the latex of Carica papaya. AB - The latex of the tropical species Carica papaya is well known for being a rich source of the four cysteine endopeptidases papain, chymopapain, glycyl endopeptidase and caricain. Altogether, these enzymes are present in the laticifers at a concentration higher than 1 mM. The proteinases are synthesized as inactive precursors that convert into mature enzymes within 2 min after wounding the plant when the latex is abruptly expelled. Papaya latex also contains other enzymes as minor constituents. Several of these enzymes namely a class-II and a class-III chitinase, an inhibitor of serine proteinases and a glutaminyl cyclotransferase have already been purified up to apparent homogeneity and characterized. The presence of a beta-1,3-glucanase and of a cystatin is also suspected but they have not yet been isolated. Purification of these papaya enzymes calls on the use of ion-exchange supports (such as SP-Sepharose Fast Flow) and hydrophobic supports [such as Fractogel TSK Butyl 650(M), Fractogel EMD Propyl 650(S) or Thiophilic gels]. The use of covalent or affinity gels is recommended to provide preparations of cysteine endopeptidases with a high free thiol content (ideally 1 mol of essential free thiol function per mol of enzyme). The selective grafting of activated methoxypoly(ethylene glycol) chains (with M(r) of 5000) on the free thiol functions of the proteinases provides an interesting alternative to the use of covalent and affinity chromatographies especially in the case of enzymes such as chymopapain that contains, in its native state, two thiol functions. PMID- 12767336 TI - Simple and unique purification by size-exclusion chromatography for an oligomeric enzyme, rat liver cytosolic acetyl-coenzyme A hydrolase. AB - An overview of the purification of an oligomeric enzyme, an extramitochondrial acetyl-coenzyme A hydrolase from rat liver, is presented. The enzyme has been purified to homogeneity using two successive size-exclusion chromatography runs, first for the monomeric and second for the oligomeric form of the enzyme. The sequential gel-filtration steps efficiently removed the contaminants of any molecular size, first of different size from that of the monomeric form of the enzyme (K(av)=0.47 on Superdex 200) and second of different size from that of the oligomeric form (K(av)=0.33), allowing us to purify the enzyme in high purity. This strategy provides an excellent model for purifying many other oligomeric proteins including key enzymes or allosteric enzymes regulating metabolism. PMID- 12767337 TI - Preparative procedures and purity assessment of collagen proteins. AB - Collagens represent a large family (25 members identified so far) of closely related proteins. While the preparative procedures for the members that are ubiquitous and present in tissues in large quantities (typically fibre and network forming collagens types I, II, III, IV and V) are well established, the procedures for more recently discovered minor collagen types, namely those possessing large non-collagenous domain(s) in their molecule, are mostly micropreparative and for some collagenous proteins even do not exist. The reason is that the proof of their existence is based on immunochemical staining of tissue slices and nucleic database searching. Methods of preparation and identification of constituting alpha-polypeptide chains as well as collagenous and non-collagenous domains are also reviewed. Methods for revealing non enzymatic posttranslational modifications (particularly of the fibre forming collagen types) are briefly described as well. PMID- 12767338 TI - Simple and rapid chromatographic purification of type V collagen from a pepsin digest of porcine intestinal connective tissue, an unmanageable starting material for conventional column chromatography. AB - A chromatographic method is described for purification of type V collagen, a minor constituent in extracellular matrix, from a pepsin digest of porcine intestinal connective tissue. The starting material was a viscous and turbid solution even after centrifugation. Direct application of the sample to a commercially available DEAE-cellulose column resulted in clogging. On the other hand, type V collagen, [alpha1(V)](2)alpha2(V) form, was successfully captured by a filter paper-based DEAE-cellulose column chromatography and purified by a subsequent commercially available cation-exchange medium without clogging. This is a vast improvement over previously described salt fractionation methods. PMID- 12767339 TI - Synthesis and chromatographic purification of recombinant human pituitary hormones. AB - Recombinant DNA-derived proteins and, in particular, human pituitary hormones, are increasingly used for research, diagnostic and therapeutic purposes. This trend has demanded new synthetic approaches and improved purification techniques. The type and sequence of the purification steps have to be selected in accordance with the cloning and protein expression strategy, the host organism and cellular localization of the protein of interest, with a view to producing the desired product at a required purity, biological activity and acceptable cost. This review article describes and analyzes the main synthetic and purification strategies that have been used for the production of recombinant human growth hormone, prolactin, thyrotropin, luteinizing hormone and follicle-stimulating hormone, giving special consideration to the few published downstream processes utilized by the biotechnology industry. Practically all types of prokaryotic and eukaryotic organisms utilized for this purpose are also reviewed. PMID- 12767340 TI - Preparative purification of B-phycoerythrin from the microalga Porphyridium cruentum by expanded-bed adsorption chromatography. AB - B-Phycoerythrin (B-PE) is a major light-harvesting pigment of microalgae. Due to its high fluorescence efficiency and its intense and unique pink color, it is widely used as a fluorescent probe and analytical reagent as well as being employed as a natural dye in foods and cosmetics. Tedious methodologies for B-PE purification have been published. In this work we present a new, fast, preparative and scaleable two-step chromatographic method for B-PE purification from the red microalga Porphyridium cruentum. Initially, phycobiliproteins were released from the microalga cells by osmotic shock and captured by applying the centrifuged cell suspension to a column containing 74 ml Streamline-DEAE equilibrated with 50 mM acetic acid-sodium acetate buffer, pH 5.5, using expanded bed adsorption chromatography at an upward flow of 200 cm h(-1). After adsorption, washing was carried out in the expanded-bed mode. Having removed unbound proteins and cellular debris, the bed was allowed to sediment and a B-PE rich solution was eluted with a downward flow of the same 250 mM buffer. In order to obtain pure B-PE, we utilized conventional ion-exchange chromatography with a column of DEAE-cellulose loaded directly with the eluate from Streamline-DEAE and developed using a discontinuous gradient of acetic acid-sodium acetate buffer, pH 5.5. With this new methodology, 66% of B-PE contained in the biomass of the microalgae was recovered, a value significantly higher than those obtained following other methodologies. The B-PE purity was tested using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and spectroscopic characterization. PMID- 12767341 TI - Endostatin capture from Pichia pastoris culture in a fluidized bed. From on-chip process optimization to application. AB - One of the characteristics of the methylothrophic yeast Pichia pastoris is its ability to grow to a very high cell density. Biomass concentrations of 300-400 g wet mass/l are common. It is therefore obvious that the recovery processes of extracellular proteins from this microorganism should take into account the effect of high biomass content. Separation by filtration and/or centrifugation is possible but these steps are cumbersome and can affect the protein recovery. The use of fluidized beds is attractive proteins capture option since it eliminates the biomass while capturing the desired protein. Zirconia-based resins possess unique properties which make them appropriate for processing high biomass concentrations in an expanded bed mode. The beads are particularly heavy (density is 3.2 g/ml) and small (75 microm) and therefore can accommodate high fluidization velocity and high mass transport. Specific operating conditions for effective capture of expressed protein have to be determined. This determination is generally time consuming and requires relatively large amount of feedstock for the lab trials. To avoid multiple chromatographic trials in columns, optimal conditions of adsorption and elution were determined by ProteinChip technology coupled with mass spectrometry. This technology involves flat chip surfaces functionalized as chromatographic beads where it is possible to adsorb and desorb proteins. Four different functional groups (strong anion-exchange, weak cation exchange, hydrophobic and metal chelate) were tested and the retained proteins were analyzed directly by mass spectrometry. The weak cation-exchange group was chosen for further work. The Zirconia-based weak cation-exchange sorbent (CM HyperZ) was evaluated for binding capacity in a packed column and then for capturing endostatin from crude feed stock. Based on the previously determined conditions; 45 l of culture containing approximately 15 kg of biomass (wet mass) and 3 g endostatin were applied on an expanded bed at a flow-rate of 535 cm/h, yielding 80% of the endostatin and removing approximately 80% of foreign proteins. PMID- 12767343 TI - Purification of a full-length recombinant glucocorticoid receptor. AB - We described a novel purification method for a recombinant glucocorticoid receptor (GR) in detail. The purification procedure consists of sequential chromatographies using common ion-exchange columns (Mono Q and Mono S). This procedure is based upon a new finding that the activated GR binds both to a Mono Q column and to a Mono S column at the same pH. The entire chromatographies took about 3 h and GR represented 97% of the purified protein sample. This purification protocol will be applicable to the purification of native GR, point mutated recombinant GR and other nuclear receptors. PMID- 12767344 TI - Enrichment of serum amyloid proteins by hydrophobic interaction chromatography combined with two-dimensional electrophoresis with immobilised pH gradients. AB - Serum amyloid A protein was subjected to one-step octyl-Sepharose extraction in three different dimensions. Elution was performed partly without UV recording, and with urea or guanidine-based buffers. The eluent was applied directly to denaturing two-dimensional electrophoresis with immobilised pH gradient, or octyl Sepharose extracted fractions were pooled and lyophilised before application. Proteins were characterised by N-terminal analysis or mass spectrometry. In most of the species that were studied, previously undescribed serum amyloid proteins were detected. Compared to conventional strategies, the presented techniques are more rational and yield more comprehensive information. The presented data also provide a basis for novel perspectives regarding certain inflammatory conditions. PMID- 12767342 TI - High-level expression of the mycobacterial porin MspA in Escherichia coli and purification of the recombinant protein. AB - MspA is the prototype of a new family of tetrameric porins and provides the main general diffusion pathway for hydrophilic compounds through the outer membrane of Mycobacterium smegmatis. Structural analysis was hampered by the scarce amount of pure protein. After replacement of the GC-rich codons of the mspA gene by codons optimal for high-level expression in Escherichia coli, the mature MspA protein was overproduced in E. coli. The recombinant MspA (rMspA) monomer (M(r) 20000) was purified by anion exchange and hydrophobic interaction chromatography yielding 2.6 mg pure protein per liter of culture. This exceeded the yield of the native protein 10-fold. Circular dichroism revealed that rMspA is folded in a native-like structure. rMspA assembled partially to the channel-forming tetramer both during expression in E. coli and after purification in vitro. Thus, overexpression in E. coli and chromatographic purification are key steps towards a high resolution structure of MspA. PMID- 12767345 TI - Preparative purification of soybean agglutinin by affinity chromatography and its immobilization for polysaccharide isolation. AB - Optimized procedures for the affinity purification of soybean agglutinin (SBA) from soybean flour, and its further immobilization, were developed. Lectin purification on galactosyl-Sepharose yielded 44.5+/-3.5 mg of pure SBA/50 g of flour. To prepare SBA adsorbents, the lectin was immobilized onto 1-cyano-4 (dimethylamino)pyridinium tetrafluoroborate (CDAP) activated Sepharose with high yields (77%). Feasibility of the use of this improved SBA adsorbent for affinity purification of Streptococcus pneumoniae capsular polysaccharides from strain 14 (CPS-14) at laboratory scale was demonstrated. Using SBA-Sepharose adsorbent (7.0 mg lectin per ml), amounts of 6.3 mg of pure CPS-14 per cycle were produced, the adsorbent being reused up to four times without loss of capacity. PMID- 12767346 TI - The mammalian XRCC genes: their roles in DNA repair and genetic stability. AB - Analysis of the XRCC genes has played an important part in understanding mammalian DNA repair processes, especially those involved in double-strand break (DSB) repair. Most of these genes were identified through their ability to correct DNA damage hypersensitivity in rodent cell lines, and they represent components of several different repair pathways including base-excision repair, non-homologous end joining, and homologous recombination. We document the phenotypic effects of mutation of the XRCC genes, and the current state of our knowledge of their functions. In addition to their continuing importance in discovering mechanisms of DNA repair, analysis of the XRCC genes is making a substantial contribution to the understanding of specific human disorders, including cancer. PMID- 12767347 TI - Decline of nuclear and mitochondrial oxidative base excision repair activity in late passage human diploid fibroblasts. AB - There are numerous studies documenting the increase of oxidative DNA damage in the nuclei and mitochondria of senescing cells as well as in tissues of aging animals. Here, we show that in IMR 90 human diploid fibroblasts, DNA repair activity is robust in both nuclear and mitochondrial extracts, however, the levels of activity differed against the three substrates tested. In extracts, cleavage of the 8-oxoguanine substrate, and to a lesser extent the dihydrouracil containing substrate, occurred in a concerted reaction between the DNA glycosylases and the second enzyme in the reaction, hAPE. Cleavage of both the furan and the dihydrouracil-containing substrates was unchanged when nuclear extracts from early and late passage cells were compared. However, cleavage of the 8-oxoguanine substrate was substantially reduced in the nuclear extracts from late passage cells and significantly reduced transcription from the hOGG1 gene was observed. When mitochondrial extracts were examined, activity on all three substrates was significantly reduced, with the reduction in hAPE activity being the most marked. The reduction in cleavage of the furan substrate was not simply due to inactive mitochondrial AP endonuclease but a substantially reduced amount of hAPE protein; transcription from the hAPE gene was also reduced. Confocal microscopic analysis confirmed that hAPE was present in the mitochondria of early passage cells but greatly reduced in the mitochondria of late passage cells. Cytoplasmic extracts from late passage fibroblasts also showed reduced activity with all three substrates suggesting that the residual hAPE, and activities that recognized dihydrouracil, were preferentially targeted to the nuclei. Taken together the data support the concept that the increase in oxidative damage in the mitochondrial DNA of senescing cells and tissues from aging animals is due to reduced base excision repair activity. PMID- 12767348 TI - The mutation frequency of 8-oxo-7,8-dihydroguanine (8-oxodG) situated in a multiply damaged site: comparison of a single and two closely opposed 8-oxodG in Escherichia coli. AB - A multiply damaged site (MDS) is defined as > or =2 lesions within a distance of 10-15 base pairs (bp). MDS generated by ionizing radiation contain oxidative base damage, and in vitro studies have indicated that if the base damage is <3bp apart, repair of one lesion is inhibited until repair of the lesion in the opposite strand is completed. Inhibition of repair could result in an increase in the mutation frequency of the base damage. We have designed an assay to determine whether a closely opposed lesion causes an increase in adenine insertion opposite an 8-oxodG in bacteria. We have positioned the MDS (an 8-oxodG in the transcribed strand and a second 8-oxodG immediately 5' to this lesion in the non-transcribed strand) within the firefly luciferase coding region. During two rounds of replication, insertion of adenine opposite the 8-oxodG in the transcribed (T) or non-transcribed (NT) strand results in a translation termination codon at position 444 or 445, respectively. The truncated luciferase protein is inactive. We have generated double-stranded oligonucleotides that contain no damage, each single 8-oxodG or the MDS. Each double-stranded molecule was ligated into the reporter vector and the ligation products transformed into wild-type or Mut Y deficient bacteria. The plasmid DNA was isolated and sequenced from colonies that did not express luciferase activity. In wild-type bacteria, we detected a translation stop at a frequency of 0.15% (codon 444) and 0.09% (codon 445) with a single 8-oxodG in the T or NT strand, respectively. This was enhanced approximately 3-fold when single lesions were replicated in Mut Y-deficient bacteria. Positioning an 8-oxodG in the T strand within the MDS enhanced the mutation frequency by approximately 2-fold in wild-type bacteria and 8-fold in Mut Y-deficient bacteria, while the mutation frequency of the 8-oxodG in the NT strand increased by 6-fold in Mut Y-deficient bacteria. This enhancement of mutation frequency supports the in vitro MDS studies, which demonstrated the inability of base excision repair to completely repair closely opposed lesions. PMID- 12767349 TI - Comparative analysis of 8-oxoG:C, 8-oxoG:A, A:C and C:C DNA repair in extracts from wild type or 8-oxoG DNA glycosylase deficient mammalian and bacterial cells. AB - We have investigated repair of DNA containing 8-oxoguanine and certain mismatches in cell-free extracts from mouse embryonic fibroblasts (MEFs) using a plasmid substrate with a single lesion at a defined position. Repair synthesis was monitored in a small restriction fragment with different size single strands in order to follow the fate of repair reactions in both strands at the same time. An important part of the study was to assess the role of OGG1 in various repair reactions and the experiments were carried out with extracts from mouse embryonic fibroblasts diploid for a mogg1 deletion (Ogg1(-/-)) as well as wild type. In wild type, DNA containing 8-oxoG:C was repaired in the expected fashion predominantly through short-patch repair. Overall repair was reduced to 20% in the Ogg1(-/-) extracts and to 40% if only long-patch repair was considered. The 8 oxoG:A pair was processed similarly in wild type and Ogg1(-/-) extracts and repair synthesis at A as well as at 8-oxoG could be demonstrated, however, to the same extent in Ogg1(-/-) and wild type for both strands. Extracts from Ogg1(-/-) behaved normally in the correction of A:C and C:C mismatches, with a strong bias for correction of A for A:C and no significant strand discrimination for C:C. Similar experiments with extracts from Escherichia coli showed a 50% reduction in the repair of 8-oxoG:C in fpg extracts and an increase to 50% above wild type in mutY. These results show that the mouse OGG1 is the major enzyme for 8-oxoG repair in the MEF cells and does not participate in mismatch repair of A:C or C:C. Furthermore, 8-oxoG opposite A appears to be repaired by a two-step repair pathway with sequential removal of A and 8-oxoG mediated by enzymes different from OGG1. PMID- 12767350 TI - The nature of DNA plays a role in chromosome segregation: endoreduplication in halogen-substituted chromosomes. AB - AA8 Chinese hamster ovary cells were treated with halogenated nucleosides analogues of thymidine, namely CldU, 5-iodo-2'-deoxyuridine (IdU), and 5-bromo-2' deoxyuridine (BrdU), following different experimental protocols. The purpose was to see whether incorporation of exogenous pyrimidine analogues into DNA could interfere with normal chromosome segregation. The endpoint chosen was endoreduplication, that arises after aberrant mitosis when daughter chromatids segregation fails. Treatment with any of the halogenated nucleosides for two consecutive cell cycles resulted in endoreduplication, with a highest yield for CldU, intermediate for IdU, and lowest for BrdU. The frequency of endoreduplicated cells paralleled in all cases the level of analogue substitution into DNA. Our results seem to support that thymidine analogue substitution into DNA is responsible for the triggering of endoreduplication. Besides, the lack of any effect on endoreduplication when CldU was present for only one S-period strongly suggest that it is the nature of template, and not nascent DNA, that plays a major role in chromosome segregation. Taking into account that topoisomerase II cleaves DNA at preferred sequences within its recognition/binding sites, the likely involvement of the enzyme is discussed. PMID- 12767351 TI - Participation of DNA polymerase II in the increased precise excision of Tn10. AB - In this work the involvement of polymerase II (Pol II) in the precise excision of Tn10 stimulated by a dnaB252 thermosensitive (Ts) mutant at the permissive temperature, by a uvrD mutant, or by mitomycin C (MMC) or ultraviolet (UV) light treatment, was investigated. A deltapolB::kan mutant showed a significant decrease in the excision of Tn10 induced by the dnaB mutation, or by MMC or UV treatment, indicating the participation of Pol II in this type of deletion process. However, no effect of Pol II was evidenced in the excision of Tn10 stimulated by the uvrD mutation. The effect of the polB mutation on Tn10 precise excision induced by all these treatments was compared to that of mutations in repair-recombination genes recF and recA. The results reveal that the degree of participation of these genes varies depending on the agent that stimulates the deletion event. PMID- 12767352 TI - The alga Chlamydomonas reinhardtii UVS11 gene is responsible for cell division delay and temporal decrease in histone H1 kinase activity caused by UV irradiation. AB - The aim of the present work was to study the possible role of the UVS11 gene of the alga Chlamydomonas reinhardtii, in regulation of the cell cycle. To characterize the defect of a uvs11 mutant in respect to DNA damage-dependent cell cycle arrest, we examined first the influence of the tubulin-destabilizing drug methyl benzimidazole-2-yl-carbamate (MBC) on inhibition of mitosis in response to UV 254nm. Then the growth and reproductive processes and activity of cyclin dependent kinases (CDK)-like kinases during the cell cycle of C. reinhardtii were investigated. In both, the wild type and the uvs11 mutant strain were compared under standard conditions and after DNA damage caused by UV 254nm. We assume the green alga C. reinhardtii possesses control mechanisms allowing to stop the cell cycle progression before mitosis in response to DNA damage. The results indicate that the uvs11 mutant is not able to stop the cell cycle after UV irradiation. We suggest that a product of the UVS11 gene affects cell response to DNA damage and influences a decrease in histone H1 kinase activity. PMID- 12767353 TI - Molecular cloning of the Drosophila Fanconi anaemia gene FANCD2 cDNA. AB - Fanconi anaemia (FA) is a rare disease characterized by chromosome instability and cancer susceptibility. With the exception of FANCD2, none of the Fanconi anaemia genes are conserved in evolution, limiting the study of the Fanconi anaemia pathway in genetically tractable models. Here we report the cloning and sequencing of a Drosophila full length cDNA homologous to human FANCD2 (dmFANCD2) as a first step in using Drosophila in Fanconi anaemia research. dmFANCD2 is composed of 14 exons coding for a protein of 1478 aminoacids. Southern blot and in situ hybridization analysis indicated that dmFANCD2 is present at single copy in the Drosophila genome and maps at the chromosomal band 92-F3. Sequence and structural biocomputational analysis indicated that, although the aminoacidic sequence, and specially the N-terminus region, is not highly conserved between humans and flies (23% identity and 43% similarity), both proteins are of the same size, globular and compact, with several transmembrane helixes and related to nuclear membrane proteins. Interestingly, the human ATM phosphorylation site at S222 and the complex-dependent monoubiquitination site at K561 are highly conserved in Drosophila at positions S267 and K595, respectively. The same is true for other putative ATM sites and their aminoacidic environment and for two out of three aminoacid mutations associated with human pathology. These results suggest that the key FANCD2 features have been conserved during over 500 million years of divergent evolution, highlighting their biological importance. PMID- 12767354 TI - DNA repair investigations using siRNA. AB - Small interfering RNA (siRNA) is a revolutionary tool for the experimental modulation of gene expression, in many cases making redundant the need for specific gene mutations and allowing examination of the effect of modulating essential genes. It has now been shown that siRNA phenotypes resulting from stable transfection with short hairpin RNA (shRNA) can be transmitted through the mouse germ line and Rosenquist and his colleagues have used shRNA, which is processed in vivo to siRNA, to create germline transgenic mice in which a target DNA repair gene has been silenced. Here, Holly Miller and Arthur P. Grollman give the background of these discoveries, provide an overview of current uses, and look at future applications of this research. PMID- 12767355 TI - 32nd annual meeting of European Environmental Mutagen Society. DNA damage and repair fundamental aspects and contribution to human disorders. PMID- 12767357 TI - Classic papers in Urology. AB - OBJECTIVE: Most clinicians are aware of the importance of using "evidence based medicine" to support their clinical practice, but where does the evidence come from? The aim of this study was to examine the concordance between papers deemed "important" by urologists compared with those selected by the more objective criteria of the citation index. METHOD: To achieve this, two approaches were used; firstly "experts" in various fields of urology were asked to select what they felt were classic papers and secondly urology journals were searched to find the 100 most cited papers for 1982-1997 and 1935-1997. The results of both of these "league tables" were then combined. RESULTS: The most cited papers varied depending on the time period studied. When the experts' selections were combined with those obtained via citation index it was found that the experts had chosen papers with a high citation index from non-urological as well as urological journals. CONCLUSION: It is possible to collate the classic papers within urology and the most effective means of doing so is to combine objective selection with expert choice. This is an exercise that can be repeated within any specialty. PMID- 12767358 TI - Artificial neural networks for decision-making in urologic oncology. AB - The authors are presenting a thorough introduction in Artificial Neural Networks (ANNs) and their contribution to modern Urologic Oncology. The article covers a description of Artificial Neural Network methodology and points out the differences of Artificial Intelligence to traditional statistic models in terms of serving patients and clinicians, in a different way than current statistical analysis. Since Artificial Intelligence is not yet fully understood by many practicing clinicians, the authors have reviewed a careful selection of articles in order to explore the clinical benefit of Artificial Intelligence applications in modern Urology questions and decision-making. The data are from real patients and reflect attempts to achieve more accurate diagnosis and prognosis, especially in prostate cancer that stands as a good example of difficult decision-making in everyday practice. Experience from current use of Artificial Intelligence is also being discussed, and the authors address future developments as well as potential problems such as medical record quality, precautions in using ANNs or resistance to system use, in an attempt to point out future demands and the need for common standards. The authors conclude that both methods should continue to be used in a complementary manner. ANNs still do not prove always better as to replace standard statistical analysis as the method of choice in interpreting medical data. PMID- 12767359 TI - Frequent use of complementary medicine by prostate cancer patients. AB - OBJECTIVES: To assess the use of complementary and alternative medicine (CAM) by prostate cancer patients and to characterize CAM-users in this population. METHODS: Prostate cancer patients who underwent routine follow-up at urological practices completed an anonymous, self-administered questionnaire on lower urinary tract function, sexuality, quality of life and use of CAM. All patients with biopsy proven cancer irrespective of age, stage and therapy were eligible. RESULTS: 822 men (69+/-8.5 years; range: 45-95) entered this survey. 245 men (29.8%) used at least one form of CAM, most frequent forms were fat-reduced diet (13.3%), selenium (10.8%) and vitamin E (9.3%). CAM-users did not differ regarding age (68.6 vs. 69.2 years), time interval between diagnosis and current evaluation (4.4 vs. 3.8 years) and PSA at diagnosis (24.9 vs. 18.5 ng/ml) from non-users. CAM-use was dependent on primary therapy (radical prostatectomy: 23.3%, radiotherapy: 31.6%, endocrine therapy: 38.8%) and disease progression. CAM-users were generally less satisfied with their treatment and rated health status and quality of life lower than those not using CAM (all p<0.001). CONCLUSIONS: CAM is used by almost 1/3 of prostate cancer patients in Austria, particularly by those with disease progression, impaired quality of life and treatment satisfaction. Urologists have to be aware of this fact and require a detailed knowledge on this issue to provide objective and evidence-based information. PMID- 12767360 TI - The role of the complexed-to-total prostate-specific antigen ratio in predicting the final pathological stage of clinically localized prostate cancer. AB - OBJECTIVES: To examine the association between the complexed-to-total (C:T) prostate-specific antigen (PSA) ratio and prostate cancer pathological stage to assess whether the C:T PSA ratio may predict the final pathological stage in patients with clinically localized prostate cancer. PATIENTS AND METHODS: In a prospective study, 101 men with clinically localized prostate cancer underwent a staging pelvic lymphadenectomy and radical prostatectomy. Total PSA (tPSA) and PSA complexed to alpha(1)-antichymotrypsin (cPSA) were measured from preoperative plasma and were correlated with the clinical and pathological stage, and with surgical margin status. The pathological stage was determined as organ-confined (n=59) and extracapsular extension (n=42). RESULTS: The distributions of tPSA and cPSA were significantly different in men with locally confined and those with locally extended disease. This finding was not observed for the C:T PSA ratio. The area under the receiver operating characteristic (ROC) curve to predict the final pathological stage was significantly greater for tPSA (0.684) and cPSA (0.677) than for the C:T PSA ratio (p<0.032). TPSA (0.685) and cPSA (0.670) also showed areas under the ROC curve greater than that of the C:T PSA ratio (0.542) (p<0.05) for prediction of positive surgical margins. CONCLUSIONS: Our results show that the C:T PSA ratio does not improve the performance of total PSA for predicting the final pathological stage in patients with clinically localized prostate cancer. PMID- 12767361 TI - Association between the clinical presentation and epidemiological features of familial prostate cancer in patients selected for radical prostatectomy. AB - OBJECTIVE: We evaluated if epidemiological features of familial prostate cancer are associated with certain clinical or histopathological characteristics of the disease. METHODS: 463 German patients with familial prostate cancer who underwent radical prostatectomy were stratified according to several epidemiological criteria: (1). the apparent mode of disease transmission, (2). the average age of onset and (3). number of affected relatives/family, (4). whether or not they met the Johns Hopkins criteria of hereditary prostate cancer. The variables analysed included the Prostate Specific Antigen (PSA) and the digital rectal examination at diagnosis, histopathological characteristics of the prostatectomy specimen and relapse free 5-year survival rates. These characteristics were compared within the subsets of familial patients and compared to 492 control patients with sporadic prostate cancer. RESULTS: Age of onset was the only clinical parameter differentiating familial and sporadic prostate cancer. Otherwise there was no association between epidemiological features of familial predisposition and the clinical presentation or outcome of the disease. CONCLUSIONS: Familial and sporadic prostate cancer seem to be the same disease. Alternatively it may be concluded that the common epidemiological features of familial prostate cancer are not useful to tell tumours that are based on inherited susceptibility apart from those that are not. Whether hereditary prostate cancer is clinically distinct from sporadic forms cannot be determined before the underlying genetic alterations are identified. PMID- 12767362 TI - Positive margins after laparoscopic radical prostatectomy: a prospective study of 100 cases performed by 4 different surgeons. AB - OBJECTIVE: Laparoscopic radical prostatectomy (LRP) has been refined by experienced surgeons into a competitive treatment alternative for localized prostate cancer. Less is known, however, about the outcomes of "learning curve" cases from newly trained surgeons. We prospectively studied 100 cases of LRP performed by 2 senior and 2 junior surgeons and addressed the rates of positive margins-an important early endpoint of oncologic efficacy. METHODS: 100 consecutive cases of LRP were performed by two senior (n=62) and two junior surgeons (n=38) by a 5-port transperitoneal route. Whole-mount step-section prostate specimens were examined by Stanford protocol. RESULTS: Positive margins occured in 25% of cases: 18% for pT2a (2/11), 18% for pT2b (11/61), 45% for pT3a (10/22), and 50% for pT3b (2/4) (p=0.002 pT2 vs. pT3). By surgeon experience, the rates were 19% (12/62) for senior and 34% (13/38) for junior (p=0.04). However, in a multiple logistic regression analysis, only pathologic stage (p=0.083) and Gleason sum (p=0.0133) reached statistical significance, while surgeon experience did not (p=0.0992). CONCLUSION: Positive margin rates after laparoscopic radical prostatectomy are significantly influenced by pathologic stage and Gleason score, and are within the range reported from open series. The higher positive margin rate from junior surgeons, although not statistically significant, suggests the need for further study and continued mentoring during surgery and/or video review of cases to improve oncologic results. PMID- 12767363 TI - Prognostic factors and survival in node-positive (N1) prostate cancer-a prospective study based on data from a Swedish population-based cohort. AB - OBJECTIVE: At presentation of prostate cancer, patients with proven lymph node metastasis (N1) are comparatively rare. It is difficult to give prognostic information based on the present literature. The aim of this study was to evaluate the impact of known risk factors in patients with pelvic node involvement and without distant metastasis. METHODS: From the population-based, prospective prostate cancer tumour registry of the South-East Region in Sweden, we collected data on all 181 patients with N1, M0 prostate cancer diagnosed from January 1987 to October 2000 with a follow-up to December 2001. Mean follow-up was 62 months. Pre-operative risk factors as age, T-category, serum PSA, tumour grade and also primary treatment given was correlated to the outcome. RESULTS: Median age at diagnosis was 65 years. Cancer-specific survival was highly variable with 5-year survival of 72%, a median of 8 years and the projected 13 year figure was 31%. T-category, age, PSA or treatment did not affect the outcome while poorly differentiated tumours had a tendency towards lower cancer-specific survival (p=0.0523) when compared to well and moderately differentiated tumours. CONCLUSIONS: This population-based cohort of prostate cancer patients with pelvic node involvement treated principally with non-curative intent had a median cancer specific survival of 8 years. Preoperatively known risk factors seem to have but a modest impact on the prognosis for patients in this stage of the disease. PMID- 12767364 TI - How important is urinary cytology in the diagnosis of urological malignancies? AB - OBJECTIVE: To audit clinical usefulness of urine cytology examination in a subspecialised urological unit setting. PATIENTS AND METHODS: Data from the hospital information support system on urinary cytology examinations carried out at one centre was audited over a period of 15 months. Source of urine cytology specimens, clinical profile of patients and the findings of urinary cytology were analysed and collated. RESULTS: A total of 1400 urinary cytology specimen on 900 patients were requested during 15 months study period. Urologists requested 1092 (78%) and non-urologists (general practitioners, physician or general surgeons) requested 318 (22%) specimens. The majority of specimens, 1115 (80%) did not show any cytological evidence of malignancy. 83 specimens (6%) showed cytological evidence of malignant cells consistent with origin from a urothelial malignancy. Among this group 87% (72) were more than 50 years of age and 60 (72%) had history of gross heamaturia. 159 (11.35) cases were reported as being suspicious of malignancy or showing atypical cells requiring further evidence. A total of 43 (3.04%) specimens were poorly preserved or insufficient for diagnosis. The positivity rate amongst urologist and non-urologists request was 56% and 6% respectively (p=0.00001 value). The source in 37 (86%) specimens reported, as poorly preserved or insufficient for diagnosis was non-urologists compared to 6 (14%) from urologists with significant p value (0.00001). CONCLUSIONS: Urinary cytology for malignant cells is a contributory investigation in the diagnosis of urological malignancy. It should be only ordered in the proper clinical situation. PMID- 12767366 TI - Sexuality preserving cystectomy and neobladder (SPCN): functional results of a neobladder anastomosed to the prostate. AB - OBJECTIVES: In order to preserve the sexual functions in patients in need of a cystectomy, a feasibility study has been performed. METHODS: In 24 male patients the seminal vesicles and the prostate were left in situ and a Studer type neobladder was anastomosed to the lateral edge of the prostate. RESULTS: Storage and voiding strongly resembled the patterns reported in neobladder patients with the anastomosis directly to the urethra. Four of the 24 males needed to perform clean intermittent catheterisation (CIC). All but one patients had daytime continence. Three patients needed a pad at night. Five patients had erectile dysfunction, of whom two responded well to sildenafil treatment, one had good rigiscan measured nightly erectile function and one had poor erections prior to the operation. Half of the patients had antegrade ejaculation, two patients reported sometimes antegrade and sometimes retrograde ejaculation. CONCLUSION: This feasibility trial showed that in the majority of our patients the remaining prostate does not interfere with micturition and the sexual functions were preserved. PMID- 12767365 TI - Alternating mitomycin C and BCG instillations versus BCG alone in treatment of carcinoma in situ of the urinary bladder: a nordic study. AB - OBJECTIVES: To evaluate whether, in patients with carcinoma in situ (CIS) of the urinary bladder, alternating instillation therapy with mitomycin C (MMC) and bacillus Calmette-Guerin (BCG) was more effective and less toxic than conventional BCG monotherapy. METHODS: Patients were stratified prospectively for primary, secondary, and concomitant CIS and randomized to one of two regimens. Patients in the alternating group received six weekly intravesical instillations of MMC 40 mg, followed by alternating monthly instillations of BCG 120 mg and MMC for one year. In the monotherapy group, only BCG was instilled on the same schedule. RESULTS: Of 323 enrolled patients, 304 were eligible for analysis. After an overall median follow-up of 56 months, the Kaplan-Meier disease-free estimate for BCG monotherapy was significantly better than that for alternating therapy (p=0.03; log rank test). Risk for progression appeared lower in the BCG monotherapy group (p=0.07), but no differences existed in survival. Besides the regimen, CIS category also predicted outcome to some extent. BCG monotherapy caused significantly more local side-effects and premature cessation of instillation treatment than did the alternating therapy. However, no differences were observed in the number of serious side-effects. CONCLUSION: One-year BCG monotherapy was more effective than the alternating therapy for reducing recurrence and compared well with the best regimens reported from substantially smaller series. The alternating therapy was better tolerated. PMID- 12767367 TI - Relative importance of sources of symptom-induced distress in urinary bladder cancer survivors. AB - OBJECTIVE: The influence of specific symptoms on emotions and social activities in the individual patient varies. Little is known about this variation in urinary bladder cancer survivors (in other words, about the relative importance of sources of symptom-induced distress). METHODS: We attempted to enroll 404 surgical patients treated with cystectomy and a conduit or reservoir in four Swedish towns (Stockholm, Orebro, Jonkoping, Linkoping), 101 surgical patients treated with cystectomy and orthotopic neobladder at the Herlev Hospital in Copenhagen, Denmark, and 71 patients treated with radical radiotherapy for bladder cancer, as well as 581 men and women controls in Stockholm and Copenhagen. An anonymous postal questionnaire was used to collect the information. RESULTS: A total of 503 out of 576 (87%) treated patients and 422 out of 581 (73%) controls participated but 59 patients were excluded. The primary source of self-assessed distress among cystectomised patients was compromised sexual function; reduced intercourse frequency caused great distress in 19% of the conduit patients, 20% of the reservoir patients and 19% of the bladder substitute patients. The primary source of self-assessed distress in patients treated with radical radiotherapy was symptoms from the bowel; 17% reported great distress due to diarrhoea, 16% due to abdominal pain, 14% due to defecation urgency and 14% due to faecal leakage. The highest proportion of subjects being distressed was 93% (substantial: 43%, moderate: 29% and little: 21%) for treated upper or lower urinary retention (indwelling catheter or nephrostomy). CONCLUSION: The distress caused by a specific symptom varies considerably and the prevalence of symptoms causing great distress differs between treatments in bladder cancer survivors. It is possible that patient care and clinical research can be made more effective by focusing on important sources of symptom-induced distress. PMID- 12767368 TI - Incidental detection beyond pathological factors as prognostic predictor of renal cell carcinoma. AB - PURPOSE: To evaluate the prognostic significance of different detection modalities of renal cell carcinoma (RCC) in a large cohort of patients who had been previously submitted to surgery in two teaching hospitals in Italy. MATERIALS AND METHODS: We reviewed the clinical records of 1446 patients who had been submitted to surgical treatment for RCC at the Departments of Urology of Padua (n=747) and Verona (n=699) from 1976 to 2000. Patients were classified into two groups according to the detection mode: symptomatic and incidental. The cancer-specific survival probability was estimated according to the Kaplan-Meier method. In order to compare the survival curves the log rank test was used. The predictive independent value of the variables was examined using the Cox proportional hazards model. RESULTS: Six hundred and thirty patients (43.6%) were treated for incidental RCC and 816 (56.4%) for symptomatic RCC. In the incidental group, the size (p<0.001), the pathological stage (p<0.001) and the nuclear grading (p<0.001) of tumors were lower than those causing symptoms. The 5-year and 10-year cancer-specific survival probability were 84% and 75% in the incidental group, and 66% and 54.5% in the symptomatic group (p<0.0001), respectively. At a multivariate analysis, the mode of detection was an independent predictive variable (H.R. 1.559), as well as pathological stage (H.R. 1.809), nuclear grading (H.R. 1.411), size 95%). The complexes [(188)Re][Re(N)(L)(2)] were characterized by chromatographic methods, and by comparison with the corresponding complexes prepared at macroscopic level starting from a non-radioactive rhenium precursor. Biodistribution studies were carried out in rats. Results showed that the complexes [(188)Re][Re(N)(L)(2)] exhibited the same biological behavior of the analogous Tc-99m complexes reported previously. The easy application of the new synthetic procedure indicates that it could be conveniently employed for preparing a large class of new Re-188 complexes having potential utilization in nuclear medicine as therapeutic agents. PMID- 12767396 TI - Decomposition of an aqueous solution of [11C]Ro 15-4513: implication of hydrated electrons in the radiolysis of [11C]Ro 15-4513. AB - An aqueous solution of [(11)C]Ro 15-4513 underwent decomposition to give a few radioactive degradation products. The major degradation product corresponded to that of authentic Ro 15-4513 with (60)Co radiolysis, which showed a molecular weight of 301 (M+H). The structure of this degradation product was estimated to have an amine group instead of an azido group on Ro 15-4513. Radiolysis of Ro 15 4513 was not suppressed by the addition of selective hydroxyl radical scavenger (MeOH, HCOO(-)), in contrast, it was suppressed effectively in the presence of a selective hydrated electron scavenger (NaNO(3)), suggesting that hydrated electrons would play an important role in the radiolysis of [(11)C]Ro 15-4513. PMID- 12767397 TI - Synthesis and biological evaluation of a fluorine-18 labeled estrogen receptor alpha selective ligand: [18F] propyl pyrazole triol. AB - The two estrogen receptor subtypes, ERalpha and ERbeta, play important roles in breast cancer. To develop an ERalpha imaging agent, we synthesized fluoropropyl pyrazole triol (FPPT, 2), an analog of our ERalpha-selective ligand PPT. FPPT retains the high ERalpha binding selectivity of its parent PPT. We prepared [(18)F]FPPT ((18)F-2) in high specific activity, but estrogen target tissue uptake in female rats was minimal and was not displaceable by unlabeled estradiol, probably because of the lipophilicity and triphenolic nature of FPPT. PMID- 12767398 TI - Synthesis and preliminary biological evaluation of radiolabeled O6-benzylguanine derivatives, new potential PET imaging agents for the DNA repair protein O6 alkylguanine-DNA alkyltransferase in breast cancer. AB - Novel radiolabeled O(6)-benzylguanine (O(6)-BG) derivatives, 2-amino-6-O-[(11)C] [(methoxymethyl)benzyloxy]-9-methyl purines ([(11)C]p-O(6)-AMMP, 1a; [(11)C]m O(6)-AMMP, 1b; [(11)C]o-O(6)-AMMP, 1c), 2-amino-6-O-benzyloxy-9-[(11)C] [(methoxycarbonyl)methyl]purine ([(11)C]ABMMP, 2), and 2-amino-6-O-benzyloxy-9 [(11)C]-[(4'-methoxycarbonyl)benzyl]purine ([(11)C]ABMBP, 3), have been synthesized for evaluation as new potential positron emission tomography (PET) imaging agents for the DNA repair protein O(6)-alkylguanine-DNA alkyltransferase (AGT) in breast cancer. The appropriate precursors for radiolabeling were obtained in two to three steps from starting material 2-amino-6-chloropurine with moderate to excellent chemical yields. Tracers were prepared by O [(11)C]methylation of hydroxymethyl or acid precursors using [(11)C]methyl triflate. Pure target compounds were isolated by solid-phase extraction (SPE) purification procedure in 45-65% radiochemical yields (decay corrected to end of bombardment), and a synthesis time of 20-25 min. The activity of unlabeled standard samples of 1-3 was evaluated via an in vitro AGT oligonucleotide assay. Preliminary findings from biological assay indicate the synthesized analogs have similar strong inhibitory effectiveness on AGT in comparison with the parent compound O(6)-BG. The results warrant further evaluation of these radiotracers as new potential PET imaging agents for the DNA repair protein AGT in breast cancer in vivo. PMID- 12767399 TI - Targeting of renal carcinoma with 67/64Cu-labeled anti-L1-CAM antibody chCE7: selection of copper ligands and PET imaging. AB - In order to optimize radiocopper labeling of anti-L1-CAM antibody chCE7, five bifunctional copper chelators were synthesized and characterized (CPTA-N hydoxysuccinimide, DO3A-L-p-isothiocyanato-phenylalanine, DOTA-PA-L-p-isocyanato phenylalanine, DOTA-glycyl-L-p-isocyanato-phenylalanine and DOTA-triglycyl-L-p isocyanato-phenylalanine). Substitution with more than 11 chelators per antibody molecule was found to influence immunoreactivity and biodistributions of (67)Cu MAb chCE7 significantly. CPTA-labeled antibody achieved the best tumor to normal tissue ratios when biodistributions of the different (67)Cu-chCE7 conjugates were assessed in tumor-bearing mice. High resolution PET imaging with (64)Cu-CPTA labeled MAb chCE7 showed uptake in lymph nodes and heterogeneous distribution in tumor xenografts. PMID- 12767400 TI - Sigma1 and dopamine D2 receptor occupancy in the mouse brain after a single administration of haloperidol and two dopamine D2-like receptor ligands. AB - We investigated sigma(1) and dopamine D(2) receptor occupancy in mouse brain after a single injection of haloperidol, nemonapride, or spiperone using [(11)C]SA4503 and [(11)C]raclopride, respectively. Co-injection of the three compounds significantly blocked the uptake of each radioligand. Six hours later, only haloperidol blocked [(11)C]SA4503 uptake, while all three reduced [(11)C]raclopride uptake. Sigma(1) receptor occupancy by haloperidol was reduced to 19% at day 2 when D(2) receptor occupancy disappeared. [(11)C]SA4503 would be applicable to the investigation of sigma(1) receptor occupancy of antispychotic drugs using PET. PMID- 12767401 TI - Synthesis and in vivo evaluation of [11C]zolpidem, an imidazopyridine with agonist properties at central benzodiazepine receptors. AB - The synthesis and evaluation of [(11)C]zolpidem, an imidazopyridine with agonist properties at central benzodiazepine receptors, is reported herein. The reaction of desmethylzolpidem with [(11)C] methyl iodide afforded the title compound [(11)C]zolpidem in a yield of 19.19 +/- 3.23% in 41 +/- 2 min in specific activities of 0.995-1.19 Ci/micromol (1.115 +/- 0.105 Ci/micromol) (n = 3; decay corrected, EOB). The amount of radioactivity in the brain after tail vein injection in male Wistar rats was low, and the regional distribution was homogeneous and not consistent with the known distribution of the central benzodiazepine receptors. The frontal cortex/cerebellum ratio was not significantly greater than one (1.007 +/- 0.266 at 5 min) and did not increase from 5 to 40 min post-injection. A PET brain imaging study in one baboon confirmed the results obtained in rats. Therefore, it can be concluded that [(11)C]zolpidem is not a suitable tracer for in vivo visualization of central benzodiazepine receptors. PMID- 12767402 TI - Kinetic modelling of [123I]CNS 1261--a potential SPET tracer for the NMDA receptor. AB - N-(1-napthyl)-N'-(3-[(123)I]-iodophenyl)-N-methylguanidine ([(123)I]CNS 1261) is a novel SPET ligand developed for imaging the NMDA receptor intra-channel MK 801/PCP/ketamine site. Data was acquired in 7 healthy volunteers after bolus injection of [(123)I]CNS 1261. Kinetic modeling showed reversible tracer binding. Arterial and venous time-activity curves overlapped after 90 min. The rank order of binding was: Thalamus > striatum > cortical regions > white matter. This distribution concurs with [(11)C]-ketamine and [(18)F]-memantine PET studies. These data provide a methodological basis for further direct in vivo challenge studies. PMID- 12767403 TI - Comment regarding Bergmann et al.'s "Assessment of the in vitro and in vivo properties of a 99mTc-labeled inhibitor of the multidrug resistant gene product of P-glycoprotein". PMID- 12767404 TI - Neyman's bias re-visited. AB - In case-control studies using prevalent cases, an apparent association may be spurious if the risk factor affects survival. In his description of this potential bias, Neyman disregarded competing risks. We use a compartment model to illustrate Neyman's bias and show that it can explain the apparent association only if the risk factor influences mortality from the disease being studied. Any effect of the risk factor on mortality from other causes is not relevant. The apparent protective effect of smoking in relation to Alzheimer disease is used as an example, but the result holds for any irreversible noncommunicable disease and for any dichotomous risk factor. PMID- 12767405 TI - Meta-analyses of molecular association studies: methodologic lessons for genetic epidemiology. AB - Meta-analyses of population-based molecular association studies have become increasingly common over the last 10 years, but little attention has been paid to methodology. In addition to the traditional considerations pertinent to any meta analysis, there are genetic issues particular to molecular association studies: checking Hardy-Weinberg equilibrium, handling data from more than two groups while avoiding multiple comparisons, and pooling data in a way that is sensitive to genetic models. We systematically reviewed all meta-analyses of molecular association studies identified via MEDLINE. Of a total of 37 studies, eight (22%) described the search terms. Nineteen (51%) did not state inclusion or exclusion criteria. Heterogeneity was assessed in 28 (76%), but only 7 of 37 (19%) studies checked for publication bias. Nine (24%) studies assessed the goodness-of-fit of Hardy-Weinberg equilibrium, and eight (22%) gave any biological rationale to justify the choice of genetic model used for pooling. There is a need for greater communication between epidemiologists and geneticists to develop methods appropriate to this area. PMID- 12767406 TI - Influence of race, clinical, and other socio-demographic features on trial participation. AB - We assessed whether sociodemographic and clinical characteristics were determinants of clinical trial participation in the Women's Estrogen for Stroke Trial (WEST) by examining data collected on women with a WEST consent visit completed after June 1996. We found no differences in consent rates by education or ethnic group (32% of white women compared with 34% of black women). Conditions associated with the intervention, such as a history of hysterectomy or previous estrogen replacement use, were strongly associated with a woman's decision to participate in this clinical trial. In multivariate models, features independently associated with consent to participate were age (odds ratio [OR] 0.96, 95% confidence interval [CI] 0.94-0.97), hysterectomy (OR 1.78, 95% CI 1.27 2.50), no cognitive impairment (OR 1.70, 95% CI 1.10-2.63), history of volunteerism (OR 1.62, 95% CI 1.02-2.63), and previous estrogen use (RR 1.58, 95% CI 1.08-2.30). PMID- 12767407 TI - Subject's Global Assessment of Relief: an appropriate method to assess the impact of treatment on irritable bowel syndrome-related symptoms in clinical trials. AB - The lack of validated outcome measures is a key limitation for the evaluation of drug efficacy in the treatment of irritable bowel syndrome (IBS). In clinical trials with tegaserod, the Subject's Global Assessment (SGA) of Relief (a global measure that includes overall wellbeing, abdominal pain/discomfort, and bowel function) was used to identify responders. A total of 1680 patients with IBS with constipation were included in two clinical studies comparing tegaserod with placebo. The SGA of Relief was obtained weekly by a single, self-administered question with five possible answers. Responders for the SGA of Relief reported statistically significant (P<.001) and clinically relevant improvements in multiple IBS-related symptoms compared with nonresponders. Response was also associated with a significant improvement in quality of life. The SGA of Relief is reliable as a new outcome measure for assessing response to therapy in IBS patients and has demonstrated responsiveness and reproducibility. PMID- 12767408 TI - A comparative review of four preference-weighted measures of health-related quality of life. AB - Four generic, preference-weighted, health-related quality of life (HRQL) questionnaires (the Quality of Well-Being, the Health Utilities Index, the EQ-5D, and the SF-6D) are reviewed. Although all of these questionnaires are designed to measure the same concept, each uses a different model of health, a different method of deriving preferences, and a different scoring formula. Head-to-head comparisons of preference-weighted questionnaires are limited. By considering several hypothetical health states, we found that utility scores for equivalent states can vary substantially, depending on the measure used. Clinicians and researchers applying preference-weighted HRQL questionnaires should be aware of such differences and exercise caution when interpreting the results. PMID- 12767409 TI - Convergence of health ratings across nephrologists, nurses, and patients with end stage renal disease. AB - We evaluated a health rating for renal failure patients that was completed by patients, nurses, and nephrologists. The study was a prospective inception-cohort follow-up design. Measurements were taken before initiating dialysis (n=206) or at the initiation of dialysis (n=200) and at 18 (n=225), 30 (n=181), 42 (n=162), 54 (n=137), and 66 (n=112) months after initiating dialysis. Patients, nurses, and nephrologists independently rated patients' health at each measurement occasion. Objective measures of health status, abstracted from the medical record, included emergency and non-emergency admissions, smoking, diabetes mellitus, pulmonary edema, history and number of myocardial infarctions (MI), basal rales, comorbid illnesses, and uremic symptoms. Simultaneous multiple regression analyses examined the correspondence between objective measures of health status and subjective health ratings separately for each rater and measurement occasion. Health ratings were averaged and submitted to the same analyses. Raters showed good agreement (average Pearson r=.43 overall), although agreement was higher between nephrologists and nurses (average r=.64) than between health professional and patients (average r=.34 and .31, respectively). All three ratings and the combined rating corresponded significantly to objective measures of health status. Uremic symptoms, emergency hospital admissions, diabetes mellitus, and recent MI correlated uniquely and most consistently with subjective health ratings. Despite overall convergence, objective measures of health status related to the groups' ratings in a complementary fashion. The health rating is reliable and relates to the current status of the patient. Performance was superior for the combined score that incorporated ratings by patients, nurses, and nephrologists. PMID- 12767411 TI - A comparison between traditional methods and multilevel regression for the analysis of multicenter intervention studies. AB - This article reviews three traditional methods for the analysis of multicenter trials with persons nested within clusters, i.e., centers, namely naive regression (persons as units of analysis), fixed effects regression, and the use of summary measures (clusters as units of analysis), and compares these methods with multilevel regression. The comparison is made for continuous (quantitative) outcomes, and is based on the estimator of the treatment effect and its standard error, because these usually are of main interest in intervention studies. When the results of the experiment have to be valid for some larger population of centers, the centers in the intervention study have to present a random sample from this population and multilevel regression may be used. It is shown that the treatment effect and especially its standard error, are generally incorrectly estimated by the traditional methods, which should, therefore, not in general be used as an alternative to multilevel regression. PMID- 12767410 TI - Some methodologic issues in analyzing data from a randomized adolescent tobacco and alcohol use prevention trial. AB - Three issues concerning the design and analysis of randomized behavioral intervention studies are illustrated and discussed within the framework of a tobacco and alcohol prevention trial among migrant Latino adolescents. The first issue arises when subjects are randomized in clusters rather than individually. Because subject observations cannot be assumed to be independent, information pertaining to the degree of clustering must be reported, and analyses must take the clustering into account. The second issue concerns the impact of compliance to the intervention and the importance of measuring compliance in the experimental and attention-control groups. A compliance analysis should control for participant contact with study personnel. Investigators must consider ways of constructing a compliance measure that is common to both conditions. Third, because outcomes are measured repeatedly over time, we illustrate the importance of assessing the impact of missing-data patterns on outcomes and the extent to which the patterns may modify the treatment effect. PMID- 12767412 TI - Improving efficiency of stroke research: the Brain Attack Surveillance in Corpus Christi study. AB - We studied whether a computer algorithm or abstractor could diagnose stroke as well as a fellowship-trained stroke neurologist. As part of an ongoing prospective, community-based stroke surveillance project, a diagnostic algorithm was developed, and patients' neurologic signs and symptoms were collected in a computerized database. The abstractors were blinded to the results of this algorithm and were asked to verify whether the patient had a stroke. The separate results of the computer and abstractor were compared with the final diagnosis given by the blinded neurologist. From 1 January through 31 July 2000, 3418 cases were screened. The abstractors yielded sensitivity 91%, specificity 97%, positive predictive value (PPV) 85%, and negative predictive value (NPV) 99%. Three computer algorithms were evaluated. The sensitivities ranged from 83% to 96%, specificity ranged from 88% to 97%, PPV ranged from 54% to 81%, and NPV ranged from 97% to 99%. The use of computer verification or abstractors may obviate the need for physician stroke verification and may greatly improve study efficiency. PMID- 12767413 TI - Improving screening accuracy for dementia in a community sample by augmenting cognitive testing with informant report. AB - This study sought to determine whether the augmentation of cognitive testing with an informant report questionnaire could improve accuracy in screening for dementia in a community setting. The sample consisted of 646 subjects aged 70-93 years. Cognitive state was assessed using the Mini-Mental State Examination (MMSE). Informants completed the 16-item short form of the Informant Questionnaire for Cognitive Decline in the Elderly (IQCODE). Dementia was diagnosed according to DSM-III-R criteria. Combination of the IQCODE with the MMSE resulted in more accurate prediction of caseness than either test alone. Receiver Operating Characteristic analysis demonstrated the superior screening performance of a logical "or" rule and a weighted sum of scores on the two tests over other methods of combination, replicating previous clinically based research. The findings also suggest that the appropriate combination of existing tests may be a fruitful method of improving screening accuracy in a variety of situations. PMID- 12767414 TI - Ginkgo for elderly people with dementia and age-associated memory impairment: a randomized clinical trial. AB - Preparations based on special extracts of the Ginkgo biloba tree are popular in various European countries. Previous studies have suggested the clinical efficacy of Ginkgo in patients with dementia, cerebral insufficiency, or related cognitive decline. However, most of these studies did not fulfill the current methodologic requirements. We assessed the efficacy of the G. biloba special extract EGb 761 in patients with dementia and age-associated memory impairment in relation to dose and duration of treatment. Our study was a 24-week, randomized, double blind, placebo-controlled, parallel-group, multicenter trial. Study participants were elderly patients with dementia (Alzheimer disease or vascular dementia) or age-associated memory impairment (AAMI). A total of 214 participants, recruited from 39 homes for the elderly in the Netherlands, were randomly allocated to Ginkgo (either 240 mg/d or 160 mg/d) or placebo (0 mg/d). After 12 weeks, the subjects in the two Ginkgo groups were randomized to continued Ginkgo treatment or placebo treatment. Primary outcome measures in this study were the Syndrome Kurz Test (SKT; psychometric functioning), the Clinical Global Impression of change (CGI-2; psychopathology, assessed by nursing staff), and the Nuremberg Gerontopsychological Rating Scale for Activities of Daily Living (NAI-NAA; behavioral functioning). One hundred twenty-three patients received Ginkgo (n=79, 240 and 160 mg/d combined) or placebo (n=44) during the 24-week intervention period. We found no statistically significant differences in mean change of scores between Ginkgo and placebo. The differences were SKT: +0.4 (90% confidence interval [CI] -0.9-1.7); CGI-2: +0.1 (90% CI -0.3-0.4), and NAI-NAA: -0.4 (90% CI -1.9-1.2). A positive difference is in favor of Ginkgo. Neither the dementia subgroup (n=36) nor the AAMI subgroup (n=87) experienced a significant effect of Ginkgo treatment. There was no dose-effect relationship and no effect of prolonged Ginkgo treatment. The trial results do not support the view that Ginkgo is beneficial for patients with dementia or age-associated memory impairment. PMID- 12767415 TI - The clinical diagnosis of acute bacterial rhinosinusitis in general practice and its therapeutic consequences. AB - In a randomized double-blind trial 251 adults with sinusitis-like symptoms were given amoxicillin/clavulanate or placebo for 6 days. Seven diagnostic indicators for acute bacterial rhinosinusitis are compared by their accuracy assuming a latent class model and by the treatment effect that they would have had if used to select a subset of patients for antibiotic treatment. Under a latent class model, radiography is a more efficient indicator then C reactive protein (CRP), which is, in turn, more efficient than other clinical signs and symptoms. However, a history of purulent nasal discharge, and signs of pus in the nasal cavity and throat, are better criteria than radiography or CRP for selecting those patients who will benefit from antibiotic treatment. These contradictory results are a salutary reminder that diagnostic indicators need to be evaluated in terms of therapeutic consequences for the patient. PMID- 12767416 TI - What proportion of rhinitis symptoms is attributable to atopy? AB - The aim of this study was to assess what proportion of patients with rhinitis symptoms (unrelated to infection) in population-based studies is attributable to atopy. We assessed the relevance of atopy in rhinitis in different countries and populations and between different age groups by performing a Medline search from 1980 onward on population-based studies. In the 22 identified population-based studies found, the overall proportion of rhinitis cases that were atopic was 61%, the proportion of noncases that were atopic was 20%, and the proportion of rhinitis cases that were attributable to atopy was 53%. There is variation between studies but little evidence of systematic geographic variation. For future research on the prevalence and possible increase of atopic rhinitis, it seems to be of major importance to differentiate between atopic and other forms of rhinitis. PMID- 12767417 TI - Effect measures in biostatistics. PMID- 12767419 TI - Comparison of usefulness of dipyridamole stress myocardial contrast echocardiography to technetium-99m sestamibi single-photon emission computed tomography for detection of coronary artery disease (PB127 Multicenter Phase 2 Trial results). AB - We hypothesized that assessment of hyperemic myocardial blood flow (MBF) velocity using myocardial contrast echocardiography (MCE) can detect coronary artery disease (CAD). We also postulated that only a single MCE study during stress is required for the detection of CAD in patients with normal function at rest. Patients with known or suspected CAD referred for dipyridamole stress technetium 99m sestamibi single-photon emission computed tomographic (SPECT) studies were enrolled. MCE was performed concurrently with SPECT using continuous infusions of PB127 during intermittent harmonic power Doppler imaging at multiple pulsing intervals. MCE and SPECT were compared in 43 of 54 patients who had adequate studies using both techniques. In 15 of the 43 patients, coronary angiography was performed within 30 days of the MCE/SPECT tests. Overall concordance for classification of patients as normal versus abnormal was 84% (kappa = 0.63) between the 2 tests. When false-negative SPECT scans were corrected for results of angiography, concordance increased to 93% (kappa = 0.82). For territorial analysis, concordance between MCE and SPECT for location of perfusion defects was 65% (kappa = 0.41) and 74% (kappa = 0.61) after SPECT was corrected by angiography. In patients with normal function at rest, a single stress MCE perfusion study allowed identification of CAD with the same concordance as rest/stress perfusion studies. In conclusion, visual assessment of regional differences in MBF velocity using PB127 allows detection of CAD with good concordance compared with technetium-99m sestamibi SPECT. In patients with normal left ventricular function at rest, a single stress PB127 MCE perfusion study is adequate for the detection of CAD. PMID- 12767420 TI - Guideline chaos: conflicting recommendations for preoperative cardiac assessment. AB - The American College of Cardiology/American Heart Association (ACC/AHA) and the American College of Physicians (ACP) have disseminated guidelines to assess preoperative cardiac risks before noncardiac surgery. The objectives of this study were to determine if these guidelines differ in preoperative recommendations for a group of patients, and whether these recommendations differ from actual provider recommendations. In this retrospective cohort study, patient characteristics and physician recommendations were abstracted from electronic medical records of consecutive patients attending a Veteran Affairs medical preoperative evaluation clinic from January 1 to April 1, 1998. Patient characteristics were used to determine what preoperative cardiac testing should have been ordered if each guideline was followed. Possible recommendations included operation without testing (OWT), noninvasive stress testing (NST), cardiac catheterization (CC), or cancel or delay surgery (OTHER). Recommendations were compared using statistical tests for agreement. Of the 138 patients identified, most underwent moderate-risk surgeries. Recommendations for preoperative testing were discordant between guidelines for 17% of patients (kappa = 0.38). Guidelines never agreed on the need for NST. Extreme differences in recommendations (i.e., one recommends OWT, the other CC) occurred in 9 patients (7%). Physicians ordered NST more often (n = 27) than either guideline. In this subgroup of patients where providers ordered a NST, the 2 guidelines significantly differed (kappa = 0.26). When applied to real patients being evaluated for surgery, ACC/AHA and ACP guidelines significantly differed in recommendations for preoperative cardiac testing. Results have implications for implementation, management, and practitioner adherence to published guidelines. PMID- 12767421 TI - Beneficial effects of rosuvastatin alone and in combination with extended-release niacin in patients with a combined hyperlipidemia and low high-density lipoprotein cholesterol levels. AB - Patients with combined hyperlipidemia and low high-density lipoprotein (HDL) cholesterol levels may benefit from combination therapy with a statin and niacin; therefore, we assessed the efficacy and safety of rosuvastatin and extended release (ER) niacin alone and in combination in 270 patients with this atherogenic dyslipidemia. Men and women > or =18 years with fasting total cholesterol levels > or =200 mg/dl, triglycerides 200 to 800 mg/dl, apolipoprotein B > or cf=110 mg/dl, and HDL cholesterol <45 mg/dl were randomized to 1 of 4 treatments in this 24-week, open-label, multicenter trial: rosuvastatin 10 to 40 mg; ER niacin 0.5 to 2 g; rosuvastatin 40 mg/ER niacin 0.5 to 1 g; or rosuvastatin 10 mg/ER niacin 0.5 to 2 g. Percent changes from baseline in low density lipoprotein (LDL) cholesterol, non-HDL cholesterol, and other lipid measurements at week 24 were determined by analysis of variance, with statistical testing performed separately between the rosuvastatin monotherapy group and each remaining treatment group. Daily doses of rosuvastatin 40 mg reduced LDL and non HDL cholesterol significantly more than either ER niacin 2 g or rosuvastatin 10 mg/ER niacin 2 g (-48% vs -0.1% and -36% for LDL cholesterol and -49% vs -11% and -38% for non-HDL cholesterol, respectively; p <0.01 for all comparisons); no additional reduction in LDL or non-HDL cholesterol was observed with the combination of rosuvastatin 40 mg/ER niacin 1.0 g (-42% and -47%; p = NS). Triglyceride reductions ranged from -21% (ER niacin monotherapy) to -39% (rosuvastatin 40 mg/ER niacin 1 g), but no observed differences were statistically significant. Compared with rosuvastatin alone, rosuvastatin 10 mg/ER niacin 2 g produced significantly greater increases in HDL cholesterol (11% vs 24%, p <0.001) and apolipoprotein A-I (5% vs 11%, p <0.017). Similar increases in HDL cholesterol and apolipoprotein A-I were noted between the monotherapy groups. Over 24 weeks, rosuvastatin alone was better tolerated than either ER niacin alone or the combinations of rosuvastatin and ER niacin. PMID- 12767422 TI - Sex and time differences in the associations of non-high-density lipoprotein cholesterol versus other lipid and lipoprotein factors in the prediction of cardiovascular death (The Rancho Bernardo Study). AB - Non-high-density lipoprotein (HDL) cholesterol (total cholesterol [TC] minus HDL cholesterol) has been suggested as the preferred lipid fraction to predict cardiovascular disease. We compared the ability of lipids, lipoproteins, the ratio of total to HDL cholesterol (TC/HDL), and non-HDL cholesterol to predict fatal coronary heart disease (CHD) and cardiovascular disease in 1,386 women and 1,094 men (mean age 69 years). After 10 years, there were more deaths in men (n = 310) than women (n = 268), but the proportions of deaths attributed to CHD (23% and 25%, respectively) and cardiovascular disease (48% and 47%) were similar. In men, age-adjusted values for non-HDL cholesterol, TC/HDL ratio, and triglycerides each predicted a significantly increased risk of CHD and cardiovascular disease; none of these associations was independent of pack-years of smoking, systolic blood pressure, fasting plasma glucose, body mass index, and physical activity. In women, age-adjusted non-HDL cholesterol levels did not predict CHD or cardiovascular disease events before or after adjusting for these covariates and for estrogen replacement therapy. In women, only the ratio of TC to HDL cholesterol predicted CHD and cardiovascular disease deaths independent of estrogen use and other risk factors. Observed associations were sensitive to time, being evident in women at 3 and 5 years, and lost thereafter, but not apparent before 10 years in men. Thus, non-HDL cholesterol is not superior to individual lipids, lipoproteins, or their ratios in the prediction of cardiovascular death in older adults. PMID- 12767423 TI - Improvement in medical risk factors and quality of life in women and men with coronary artery disease in the Multicenter Lifestyle Demonstration Project. AB - This study examined medical and psychosocial characteristics of 440 patients (mean age 58 years, 21% women) with coronary artery disease at baseline and at 3 month and 12-month follow-ups. All patients were participants in the Multicenter Lifestyle Demonstration Project, aimed at improving diet (low fat, whole foods, plant-based), exercise, stress management, and social support. Spousal participation was encouraged. Both genders evidenced significant improvements in their diet, exercise, and stress management practices, which they maintained over the course of the study. Both women and men also showed significant medical (e.g., plasma lipids, blood pressure, body weight, exercise capacity) and psychosocial (e.g., quality of life) improvement. Despite their worse medical, psychosocial, and sociodemographic status at baseline, women's improvement was similar to that of men's. These results demonstrate that a multi-component lifestyle change program focusing on diet, exercise, stress management, and social support can be successfully implemented at hospitals in diverse regions of the United States. Furthermore, this program may be particularly beneficial for women with coronary artery disease who generally have higher mortality and morbidity than men after a heart attack, angioplasty, or bypass surgery. PMID- 12767424 TI - Performance of an automatic external cardioverter-defibrillator algorithm in the discrimination of supraventricular from ventricular tachycardia. AB - An automatic external cardioverter-defibrillator (AECD) with a programmable supraventricular-ventricular tachycardia (VT) zone underwent evaluation of arrhythmia discrimination performance in the electrophysiologic laboratory during induced supraventricular tachycardia (SVT) and unipolar and bipolar atrial pacing. The AECD SVT zone was programmed so that the induced SVT rate would fall within this zone. Atrial pacing was also performed at a rate within this zone. The ability of the AECD to accurately discriminate between VT and SVT and to recommend shock delivery was assessed. A total of 98 patients underwent conventional diagnostic electrophysiologic studies (49 men, age 59 +/- 19 years) with a total of 55 inducible sustained SVTs. High right atrial pacing was performed in 56 patients in unipolar and in 82 patients in bipolar fashion. In response to induced sustained SVT, the AECD correctly classified 47 episodes as nonshockable, 4 incorrectly as shockable, and 4 episodes correctly as shockable with a resultant sensitivity of 100% and specificity of 92%. Bipolar high right atrial pacing was correctly identified as nonshockable in 75 episodes, incorrectly identified as shockable in 5 episodes, and correctly identified as shockable in 2 episodes with a resultant sensitivity of 100% and specificity of 94%. The Powerheart AECD accurately discriminates SVT from VT and is expected to correctly deliver automatic external shocks rapidly in the presence of spontaneous life-threatening tachycardia and appropriately withhold therapy during SVT. PMID- 12767425 TI - Assessment of left ventricular diastolic function by radionuclide ventriculography at rest and exercise in subclinical hypothyroidism, and its response to L-thyroxine therapy. AB - Hypothyroidism is associated with intrinsic myocardial changes reflected by alterations in contractility and relaxation. Diastolic function, however, rather than systolic cardiac function, seems to be mostly impaired by thyroid hormone deprivation. Our aim was to evaluate diastolic function at rest and during maximal exercise by means of radionuclide ventriculography in subclinical hypothyroidism before and after restoration of euthyroidism. Ten subclinical hypothyroid patients (50 +/- 8.7 years) (thyroid-stimulating hormone 11 +/- 4.2 microUI/ml) without cardiac disease were studied before and 6 months after levothyroxine (L-T(4)) replacement (thyroid-stimulating hormone 1.9 +/- 1.1 microUI/ml). We compared the basal and post-therapy cardiac parameters with a control group of 14 euthyroid patients (52.5 +/- 10 years) (thyroid-stimulating hormone 2.5 +/- 1.2 microUI/ml). Multigated equilibrium radionuclide ventriculography was performed to assess systolic and diastolic ventricular function. Student's t and paired Student's t tests were applied for statistical analysis. We found a significant difference between the time to peak filling rate (TPFR) at rest before (0.241 +/- 0.002 ms) and after (0.190 +/- 0.012 ms) treatment with L-T(4). A significant difference that disappeared after restoration of euthyroidism was also observed between the basal TPFR values of the subclinical hypothyroid patients and the control group (0.189 +/- 0.01 ms). The same pattern was observed at maximal exercise. Thus, TPFR, a parameter of left ventricular (LV) diastolic function measured by radionuclide ventriculography, is impaired in subclinical hypothyroid patients both at rest and during exercise and returns to normal values after L-T(4) therapy. PMID- 12767426 TI - Thyroid hormone and cardiac contractility. PMID- 12767427 TI - Multiple versus single coronary plaque ruptures detected by intravascular ultrasound in stable and unstable angina pectoris and in acute myocardial infarction. PMID- 12767428 TI - Determinants of angiographically silent stenoses in patients with coronary artery disease. PMID- 12767429 TI - Comparison of white blood cell counts in acute myocardial infarction patients with significant versus insignificant coronary artery disease. PMID- 12767430 TI - One-year results of percutaneous myocardial revascularization for refractory angina pectoris. PMID- 12767431 TI - Comparison of effects of abciximab versus eptifibatide on C-reactive protein, interleukin-6, and interleukin-1 receptor antagonist after coronary arterial stenting. PMID- 12767432 TI - Effects of coadministration of aspirin or clopidogrel on exercise testing in patients with heart failure receiving angiotensin-converting enzyme inhibitors. PMID- 12767433 TI - Comparison of characteristics of patients with coronary heart disease receiving lipid-lowering therapy versus those not receiving such therapy. PMID- 12767434 TI - Effect of atorvastatin (80 mg) on recurrent ischemia in unstable angina pectoris or non-ST-elevation acute myocardial infarction. PMID- 12767435 TI - Relation of electrocardiographic abnormalities to levels of serum C-reactive protein. PMID- 12767436 TI - Relation of high lipoprotein(a) to other traditional atherosclerotic risk factors in patients with coronary heart disease. PMID- 12767437 TI - Effect of systemic immunosuppression on coronary in-stent intimal hyperplasia in renal transplant patients. PMID- 12767438 TI - Prolonged postischemic regional left ventricular delayed relaxation or diastolic asynchrony detected by color kinesis following coronary vasospasm. PMID- 12767439 TI - Comparison of the effects of dobutamine and nesiritide (B-type natriuretic peptide) on ventricular ectopy in acutely decompensated ischemic versus nonischemic cardiomyopathy. PMID- 12767440 TI - Causes of impaired consciousness while driving a motorized vehicle. PMID- 12767441 TI - Etiologies of left bundle branch block and correlations with hemodynamic and angiographic findings. PMID- 12767442 TI - Prevalence and correlates of rheumatic heart disease in American Indians (the Strong Heart Study). PMID- 12767443 TI - Cardiac risk in pregnant women with rheumatic mitral stenosis. PMID- 12767444 TI - Early and intermediate-term outcomes of pregnancy with congenital aortic stenosis. PMID- 12767445 TI - Frequency of development of idiopathic dilated cardiomyopathy among relatives of patients with idiopathic dilated cardiomyopathy. PMID- 12767446 TI - Comparison of the effects of pentaerythritol tetranitrate and nitroglycerin on endothelium-dependent vasorelaxation in male volunteers. PMID- 12767447 TI - Prenatal diagnosis and in utero treatment of torsades de pointes associated with congenital long QT syndrome. PMID- 12767448 TI - Liver dysfunction and heart failure. PMID- 12767450 TI - Childhood disintegrative disorder. AB - In 1908 a Viennese remedial educator Theodor Heller described six children under the name of dementia infantilis who had insidiously developed a severe mental regression between the 3rd and 4th years of life after normal mental development. Neuropathological and other medical conditions are sometimes associated with this disorder, but contrary to earlier belief this is not typical. Interest in childhood disintegrative disorder has increased markedly in recent years and in this review attention is given to more recently published cases based on ICD-9, ICD-10 and DSM-IV diagnostic systems. Information is provided related to nosology, epidemiological data, differential diagnosis, aetiology, treatment and outcome. PMID- 12767451 TI - Multiple sclerosis in children. AB - Multiple sclerosis (MS) is traditionally the domain of adult neurologists due to its characteristic presentation during early adult life. Although descriptions of infants with MS appeared in the beginning of the last century and the first autopsy was described even earlier, it was not until 1980 that childhood onset MS was recognized and subsequently well characterized. In spite of this, the awareness of pediatricians and pediatric neurologists to the occurrence of MS especially in infants and young children is still unsatisfactory. It is not infrequent that a meticulous, time consuming and costly search for metabolic and degenerative disorders other that MS is initiated before the diagnosis of MS is considered. This leads to a significant diagnostic and therapeutic delay in many young patients. Moreover, when the presentation is acute and characterized by confusion, seizures, CSF pleocytosis following a viral infection, a diagnosis of meningoencephalitis will be frequently reached. In this review, updated data on frequency, epidemiology, some special clinical and radiological features of childhood onset MS, outcome and treatment will be briefly discussed with the purpose of alerting physicians to the possibility of the occurrence of MS even in infants and young children. PMID- 12767452 TI - Childhood and juvenile onset multiple sclerosis: clinical and paraclinical features. AB - It is well known that multiple sclerosis (MS) is a demyelinating disease of the central nervous system that mostly starts in the second to third decade. In the present study, we reviewed our own observations of the clinical and paraclinical features in the 36 of 890 (4.04%) MS patients whose symptoms started before 16 years of age. The average age at onset of the disease in these 36 patients was 12.9 years. In 18 patients, the disease onset was monosymptomatic. Diplopia and sensory disturbances were the most common initial manifestations and occurred in 27.7% of cases. Twenty-one patients (59%) had a relapsing and 11 patients (30.5%) had a secondary progressive course. On the last evaluation, the EDSS score was above 5 in 11 patients and it was below 5 in 21 patients. As a result of this study, we concluded that childhood onset MS does not significantly differ from that it has been typically seen in adults in terms of major clinical manifestations and course of disease. PMID- 12767453 TI - Idiopathic stabbing headache: clinical characteristics of children and adolescents. AB - Very few cases of short-lasting headaches have been reported in children and adolescents. The aim of this study is to describe a group of patients in the pediatric age group with short attacks of head pain and to demonstrate that they are not easily classified according to existing criteria for adults. We describe 23 subjects in retrospect, 12 males and 11 females, with brief attacks of headache, stabbing in nature, self-limited, lasting from a few seconds to 15 min. This sample was taken from a total population of 548 children and adolescents referred to our Pediatric Headache Unit during the years 1995-1999. Mean age at onset of symptomatology was 9 years. The localization of the headache was bilateral in 60% of the patients and unilateral in 40%. The pain was either unifocal at the orbit or temple (60%) or multifocal (40%). In 24% of the children, the interictal awake EEG showed infrequent posterior slow-waves. In 12 patients we used paracetamol with a good response. Although these painful episodes shared some aspects with the adult form of idiopathic stabbing headache, they had different durations of attack, and other primary headache syndromes did not accompany them either at the time of presentation or during the following 2 years. Short attacks of headache are present in the pediatric age group and are not easily classified according to the International Headache Society criteria. Their nature and correlation with migraine remains unclear. We propose follow-up of these subjects to obtain a better description of the natural history of these forms of headache in the pediatric age group. PMID- 12767454 TI - An MEG study of P300 activity during a color discrimination task 2: source localization study. AB - In this study, we described developmental changes in an event-related magnetic source of P300 activity during a visual oddball paradigm using magnetoencephalography. The maximum and minimum points of each peak of target P300 activity were scattered in a more anterior temporal area in children than in adults. The single dipole source analysis successfully estimated a dipole in the vicinity of the thalamus and the cingulate gyrus in adults. The location differed in children, although the dipole had overall greater moment in children than in adults. Our findings suggest that the thalamus plays an important role in generating P300 activity specific to color discrimination processing, and that in children the source is not consistent during development. PMID- 12767455 TI - Leigh syndrome associated with West syndrome. AB - Leigh syndrome (LS) (sub-acute necrotizing encephalomyelopathy) is characterized by symmetric brain lesions occurring mainly in the basal ganglia and associated with variable clinical manifestations such as hypotonia, psychomotor retardation, and feeding difficulties. Patients with LS may develop seizures. Only three patients with LS have been identified in the literature as having West syndrome (WS). We have seen 12 children with LS in the past 20 years, and noticed that as many as five of them developed WS. This report discusses five LS children with WS, comparing them with seven LS children without WS. In all five patients, infantile spasms developed after LS had become evident, in addition to other type(s) of seizures. The onset of LS in all the patients with WS was before 10 months of age. Although not statistically proven, early onset of LS, spasticity, nystagmus, apnea, poor feeding, and cardiac problems seemed to be associated with the development of WS. We were not able to conclude that certain types of symptoms or examination results of patients with LS indicated the development of WS. The association of LS with WS did not markedly influence the prognoses of the children. WS may not be a rare complication of LS, especially in infants under 12 months of age. This report is the first review of LS associated with WS. PMID- 12767457 TI - Visual agnosia in a child with non-lesional occipito-temporal CSWS. AB - In this paper we describe a case of severe visual agnosia in a child with an electrophysiological pattern of continuous spike-wave discharges in slow sleep (CSWS) in the occipito-temporal regions. The neuropsychological spectrum related to this phenomenon is discussed. Published paediatric reports associate visual agnosia (i.e. an inability to recognize objects without impairment of visual acuity) mainly with symptomatic occipito-temporal aetiology (e.g. cortical dysplasia, vascular insults) and other neurological symptoms (e.g. autism). We describe a detailed 2 year electrophysiological and neuropsychological follow-up of an 8-year-old boy with sporadic seizures, occipito-temporal CSWS and visual agnosia. The growth and neurological development of the child had been considered as normal, neurological examination did not reveal any focal signs, visual acuity was intact and MRI was normal. First EEG and six consecutive 24 h video EEG recordings during the follow-up of 22 months showed continuous spike-and-wave activity covering over 85% of the non-REM sleep. According to structured neuropsychological tests (Wechsler Intelligence Scale for Children--Third Edition, A Developmental Neuropsychological Assessment (NEPSY), Test of Visual Perceptual Skills, Corsi block, Hooper Visual Organization Test) the boy had normal verbal intelligence but major deficits in visual perception, especially in object recognition, impaired shape discrimination and detection, and poor copying skills. Attention and executive functions were intact. There were no difficulties in short- or long-term memory. Verbal cues and naming the objects improved visual memory. Tracing the objects with a finger or by moving the head improved object recognition. Currently the boy attends a special school with a rehabilitation plan including neuropsychological and occupational therapies. This case adds a new facet to the spectrum of neuropsychological deficits in children with CSWS. Sleep EEG should be included in the etiological studies of children with specific neuropsychological problems and detailed neuropsychological assessment is needed for diagnostic and rehabilitation purposes. PMID- 12767456 TI - Can memory be improved? A discussion on the role of ras, GABA, acetylcholine, NO, insulin, TNF-alpha, and long-chain polyunsaturated fatty acids in memory formation and consolidation. AB - It is proposed that long-chain polyunsaturated fatty acids when given from the perinatal period will ensure proper development and growth of the brain and maintain the activity and/or concentrations of ras, nitric oxide, insulin, and various neurotransmitters and cytokines at physiological level and thus, improve memory and prevent learning deficits. PMID- 12767458 TI - Visual evoked potentials in children with occipital epilepsies. AB - The objectives of this study are to see if any visual evoked potential (VEP) differences are present in two forms of occipital epilepsy, childhood epilepsy with occipital paroxysms (CEOP) and symptomatic occipital epilepsy (SOE) with respect to etiology, as CEOP is a benign age- and localization-related idiopathic epilepsy while SOE is a symptomatic form. Nineteen patients with CEOP and 13 patients with SOE were included in the study and P100 potential latency and amplitude values obtained from these patients were compared with the values recorded from normal controls. The amplitude values recorded from the patients with CEOP were significantly high (P=0.033). P100 potential latency values recorded in patients with SOE were significantly long (P=0.028). High amplitude VEP responses were mostly attributed to hyperexcitability of the occipital cortical structures whereas prolonged latency P100 responses were attributed to occipital structural changes. PMID- 12767459 TI - Serum neuron-specific enolase in children with febrile seizures: time profile and prognostic implications. AB - Compared to publications of elevated levels of neuron-specific enolase (NSE) in adult patients with single seizures or epilepsy, data in children are rare. We studied serial NSE serum concentrations in children after febrile convulsions (FC). In addition, the predictive value of NSE levels in serum for recurrence of FC or further development of epilepsy was determined. Serum NSE levels were determined at (1) 0-2 h, (2) 6-8 h and (3) 20-24 h after a first or second FC in children aged 4 months to 6 years. Eighty-two patients (35 female, 47 male) aged four months to 5.7 years were included. Seventy-one children had generalized, and seven focal FC. The seizures in the remaining four patients could not be properly classified. During the follow-up of 14-28 months 13 patients had at least one more FC and in five epilepsy due to recurrent afebrile seizures was diagnosed. There was no statistically significant elevation of NSE concentration in the group of children with FC or the group with recurrent FC or epilepsy. The comparison of the NSE values at different times after FC did not show any significant differences either. It seems from our results that NSE activity cannot be used as a predictor for possible brain damage caused by FC and that it is not of predictive value considering further FC or development of epilepsy. We cannot confirm the published results of the elevation of NSE serum levels in adults with single seizures or status epilepticus. PMID- 12767460 TI - Buccal midazolam for treatment of prolonged seizures in children. AB - Midazolam is a relatively new anticonvulsive agent in the benzodiazepine group. It has a short onset of duration and is practical for use, providing several alternatives such as intravenous, intramuscular, and intranasal routes. The buccal route could be an alternative choice for seizure control in an emergency setting. However, no sufficient reports are available on buccal midazolam administration. The present study was designated to examine the efficacy of buccal midazolam in children at different ages with seizures of more than 5 min duration. Nineteen previously unreported children, aged from 1 month to 15 years, were treated with a 0.3 mg/kg dose of buccal midazolam; 13 had prolonged seizures, and six had status epilepticus, with a duration of 5-45 min (mean 22 min). Sixteen of 19 seizures (84.2%) stopped within 10 min of buccal midazolam being given. The drug efficacy in patients with status epilepticus was 50%. However, all patients with convulsions shorter than 30 min showed a perfect response (100%). Convulsion episodes stopped within 3.89+/-2.22 min (median time 3 min). Seizure duration was correlated with cessation of seizure (r=0.76, P<0.001). No clinically important side effects were seen in any patient. On the basis of this experience, we concluded that a 0.3 mg/kg dose of buccal administration of midazolam might offer an effective treatment in all ages of children. PMID- 12767461 TI - A case of subacute sclerosing panencephalitis preceded by epileptic seizures: evolutional EEG changes. AB - We reported a patient with subacute sclerosing panencephalitis (SSPE) in whom EEG had been serially performed before the onset. She was referred to our hospital due to epileptic seizures at 2 years of age. Focal spikes were seen on EEG at the time of her first seizure (2 years 4 months). At the time of the second unprovoked seizure (9 years 8 months), EEG showed poorly organized background activity associated with focal spikes and a few diffuse spike-and-waves. Then, diffuse paroxysms became predominant, followed by periodic synchronous discharges. In our case, EEG abnormalities were recognized before mental deterioration. Unexpected EEG changes in a patient with epilepsy could be a clue as to the diagnosis of SSPE. PMID- 12767462 TI - A study of EEG and epilepsy profile in Wolf-Hirschhorn syndrome and considerations regarding its correlation with other chromosomal disorders. AB - Wolf-Hirschhorn syndrome (WHS) is a genetic disorder caused by a deletion of the short arm of chromosome 4. Sgro et al. described an electroclinical profile for WHS, but data regarding this issue are scarce. We report an 8-year-old girl presenting the classic phenotype for WHS, confirmed by FISH test. Epilepsy started during infancy with myoclonic seizures. Later, she presented atypical absences, which gradually increased in frequency, and at the age of 2.5 years, she presented a non-convulsive status epilepticus. Epilepsy was controlled with valproate at the age of 6 years. Serial EEGs were performed and showed unusual bursts of generalized, high amplitude delta waves with superimposed low-moderate amplitude sharp waves. A literature review was performed and our case was compared to others, where EEG and/or epilepsy were addressed. Our case and previously published data show that WHS presents a stereotyped epilepsy profile and EEG patterns. A discussion concerning similarities between these findings and those observed in Angelman syndrome has been performed, since in both syndromes, GABA genes are involved and may play a role in the pathogenesis. Although fascinating, this theory is simplistic, since patients with Angelman syndrome without GABA deletion may present epilepsy and EEG abnormalities. Another issue is the striking overlap regarding these features, between WHS and Pitt-Rogers Danks syndrome, which may be a key in showing that these disorders could be a spectral variation of the same entity. PMID- 12767463 TI - Familial moyamoya disease in a Greek family. AB - Moyamoya disease (M-M) is characterized by progressive obstruction of the supraclinoid portion of internal carotid arteries and the proximal middle, anterior and posterior cerebral arteries, associated with the formation of a characteristic net of collateral vessels in the basal ganglia region. Clinical manifestations in childhood include transient ischaemic attacks, seizures and multiple infarcts. Approximately 7% of M-M cases are familial. We report two affected Greek siblings with typical clinical and neuroradiological findings of M M. Linkage analysis of the whole family was consistent with linkage to the region 3p24-26, as previously reported in other familial Japanese M-M cases. PMID- 12767464 TI - Neuropsychological developmental change in a case with Noonan syndrome: longitudinal assessment. AB - The authors describe the neuropsychological development of a 10-year-old boy with Noonan syndrome. The subject's IQ showed normal intelligence, although there was a discrepancy between verbal and performance IQs. Visual perception was delayed, with clumsiness and inattention affecting his performance. Both visual perception and hyperactivity improved with the subject's general development, but his inattention seemed to increase after the age of 9. The behavioral features and cognitive profile of our case resembled those of attention-deficit/hyperactivity disorder. We recommend that clinicians should evaluate cautiously the specific profile of cognitive development in Noonan syndrome with particular focus on controlling the patient's inattention. PMID- 12767465 TI - Schinzel-Giedion syndrome: a further cause of West syndrome. AB - Schinzel-Giedion syndrome (SGS) is a rare disorder with a likely autosomal recessive pattern of inheritance which is characterized by several facial dysmorphisms, midface hypoplasia, multiple skeletal anomalies including short and sclerotic skull base, short neck, and post-axial polydactyly. Cardiac and urogenital malformations are also present. Thirty-three cases have been described so far. We report on a boy affected by SGS in whom a long-term EEG follow-up showed a progressive deterioration of the background bioelectric activity ending, at the age of 19 months, with a hypsarrhythmic pattern clinically correlated with severe and refractory infantile spasms. EEG deterioration and neuroradiological findings, which showed progressive brain atrophy, confirm the neurodegenerative nature of SGS. We also re-evaluated all the published cases and found that 33% of patients with SGS experienced neonatal seizures and another 25% developed West syndrome in the following months. The seizures appeared extremely refractory to several anticonvulsive treatments. In conclusion, we believe that SGS should be included among the causes of secondary West syndrome. PMID- 12767466 TI - Mitochondrial dysfunction and nucleoside reverse transcriptase inhibitor therapy: experimental clarifications and persistent clinical questions. AB - Nucleoside reverse transcriptase inhibitors (NRTIs) in combination with other antiretrovirals (HAART) are critical in current AIDS therapy, but mitochondrial side effects have come to light with the increased use of these compounds. Clinical experience, pharmacological, cell and molecular biological evidence links altered mitochondrial (mt-) DNA replication to the toxicity of NRTIs in many tissues, and conversely, mtDNA replication defects and mtDNA depletion in specific target tissues are observed. The shared features of mtDNA depletion and energy depletion became key observations and related the clinical and in vivo experimental findings to inhibition of mtDNA replication by NRTI triphosphates in vitro. Subsequent to those findings, other observations suggested that mitochondrial energy deprivation is concomitant with or the result of mitochondrial oxidative stress in AIDS (from HIV, for example) or from NRTI therapy itself. With increased use of NRTIs, mtDNA mutations may become increasingly important pathophysiologically. One important future goal is to prevent or attenuate the side effects so that improved efficacy is achieved. PMID- 12767467 TI - An in vitro system combined with an in-house quantitation assay for screening hepatitis C virus inhibitors. AB - Hepatitis C virus (HCV) infection is a serious global health problem. Interferon alpha (IFN-alpha) and ribavirin have demonstrated efficacy in the treatment of HCV infection; however, these therapies display many side effects. To screen the anti-HCV compounds from plants, we established an in vitro model for inoculation of HCV by centrifugation-facilitated method. The HCV RNA molecules were then quantitated by nested competitive reverse transcription-polymerase chain reaction (cRT-PCR) using fluorescein-labeled primers, and analyzed by ABI Prism 310. The positive and negative strands of HCV RNA were detectable in Vero cells on Day 7 post-infection, suggesting that the HCV RNA was present in the cell model system. The cell culture system was further used to screen the anti-HCV activities of 4 Chinese herbal formulas and 15 formula components. IFN-alpha showed an antiviral effect. The formulas exhibited no anti-HCV activities, while Arnebia euchroma, Thlaspi arvense, and Poncirus trifoliata displayed anti-HCV activities. Therefore, these results pointed out the possibility by using the cell culture system established in this study to screen the herb extracts for their anti-HCV activities. PMID- 12767468 TI - Lactoferrin and lactoferricin inhibit Herpes simplex 1 and 2 infection and exhibit synergy when combined with acyclovir. AB - Lactoferrin (LF) is a multifunctional glycoprotein, which plays an important role in immune regulation and defense mechanisms against bacteria, fungi, and viruses. Upon peptic digestion of LF, a peptide called lactoferricin (Lfcin) is generated. Lfcin corresponds to the N-terminal part of the protein. In this study we investigated the antiviral activity of bovine and human Lfcin against Herpes simplex virus (HSV)-1 and HSV-2. The 50% effective concentrations (EC(50)) for LF and Lfcin against several clinical isolates of HSV-1 and HSV-2, including acyclovir (ACV)-resistant strains, were determined. We further evaluated the effect of the combination of either LF or Lfcin with ACV against HSV-1 and HSV-2. Synergy was observed between both LF or Lfcin in combination with ACV against the HSV laboratory strains. The 50% effective concentration (EC(50)) for ACV and LF or Lfcin, when combined with ACV, could be reduced by two- to sevenfold compared to the EC(50) when the drugs were used alone. PMID- 12767469 TI - Evaluation of mitochondrial DNA content and enzyme levels in tenofovir DF-treated rats, rhesus monkeys and woodchucks. AB - The antiviral compound tenofovir DF (Gilead Sciences) was evaluated for possible mitochondrial toxicity in rats, rhesus monkeys and woodchucks. Animals were treated by oral gavage with tenofovir DF, and the levels of mitochondrial enzymes cytochrome c oxidase and citrate synthase were assayed. In rats (6/group) treated daily for 28 days with 300 mg/kg tenofovir DF the enzyme levels were unchanged versus control in liver, kidney, and skeletal muscle. In a parallel study, rats (6/group) were treated with 40 mg/kg of the antiviral adefovir dipivoxil (Gilead Sciences) and enzyme levels were also unchanged versus control. In rhesus monkeys (6/group) treated daily with 30 mg/kg or 250 mg/kg tenofovir DF for 56 days, and in woodchucks (6/group) treated daily with 15 mg/kg or 50mg/kg tenofovir DF for 90 days, the enzyme levels were unchanged in liver, kidney, skeletal muscle and cardiac muscle. Mitochondrial DNA (mtDNA) content was determined in tissue from treated versus control animals by utilizing a quantitative real-time PCR (QPCR) technique, where the relative ratios of mitochondrial cytochrome b gene to the genomic actin gene were measured. The relative mtDNA content from rats, rhesus monkeys and woodchucks were unchanged in the various treatment groups. Variations in mtDNA content between animals in the same treatment group were noted. The actual species-dependent mitochondria/genomic ratios were estimated from the QPCR assay. In summary, treatment with tenofovir DF, or with adefovir dipivoxil, did not affect mtDNA content or level of mitochondrial enzymes, and no liver, muscle or renal microscopic abnormalities were observed in tenofovir-treated animals. PMID- 12767470 TI - Metabolic transformation of AZTp4A by Ap4A hydrolase regenerates AZT triphosphate. AB - The reverse transcriptase (RT) of HIV which has been inhibited by the incorporation of AZT into the primer strand is subject to a deblocking reaction by cellular ATP. This reaction yields unblocked primer plus the dinucleoside tetraphosphate, AZTp(4)A. In the present study, we report that AZTp(4)A is an excellent substrate for the enzyme Ap(4)A hydrolase (asymmetrical dinucleoside tetraphosphatase, EC 3.6.1.17), an enzyme that is widely distributed in many cell types. Progress of the reaction has been monitored by 31P NMR, and it was found that hydrolysis results in the production of AZTTP:ATP in a 7:1 ratio. The AZTp(4)A was also hydrolyzed at a rate 1.8-fold more rapidly than Ap(4)A. Spectrophotometric assays yielded Michaelis constants of 2.35 and 0.71 microM for Ap(4)A and AZTp(4)A, respectively. It, therefore, appears that Ap(4)A hydrolase can play a useful role in the regeneration of the AZTTP, the active form of AZT, for the inhibition of HIV RT. PMID- 12767471 TI - Elucidation of HIV-1 protease resistance by characterization of interaction kinetics between inhibitors and enzyme variants. AB - The kinetics of the interaction between drug-resistant variants of HIV-1 protease (G48V, V82A, L90M, I84V/L90M, and G48V/V82A/I84V/L90M) and clinically used inhibitors (amprenavir, indinavir, nelfinavir, ritonavir, and saquinavir) were determined using biosensor technology. The enzyme variants were immobilized on a biosensor chip and the association and dissociation rate constants (k(on) and k(off)) and affinities (K(D)) for interactions with inhibitors were determined. A unique interaction kinetic profile was observed for each variant/inhibitor combination. Substitution of single amino acids in the protease primarily resulted in reduced affinity through increased k(off) for the inhibitors. For inhibitors characterized by fast association rates to wild-type protease (ritonavir, amprenavir, and indinavir), additional substitutions resulted in a further reduction of affinity by a combination of decreased k(on) and increased k(off). For inhibitors characterized by slow dissociation rates to wild-type enzyme (saquinavir and nelfinavir), the decrease of affinity conferred by additional mutations was attributed to increased k(off) values. Development of resistance thus appears to be associated with a change of the distinctive kinetic parameter contributing to high affinity. Further inhibitor design should focus on improving the "weak point" of the lead compound, that being either k(on) or k(off). PMID- 12767472 TI - Canonical 3'-deoxyribonucleotides as a chain terminator for HCV NS5B RNA dependent RNA polymerase. AB - Nucleoside chain terminators represent one of the most promising classes of antiviral drug for DNA viruses and retroviral infection; however, they have not been fully explored against RNA viral polymerases. In this report, we investigate the notion of employing canonical 3'-deoxyribonucleoside triphosphates (3'-dNTPs) as a chain terminator for hepatitis C virus (HCV) NS5B RNA-dependent RNA polymerase (RdRp). Using a HCV RNA transcript-dependent RNA elongating assay, we found that they inhibit NS5B RdRp with K(i) ranged from 0.7 to 23 microM. Additional structure-activity relationship studies showed that removal of 2' hydroxyl group, elimination of ribose's 2',3'-carbon-carbon bond, or addition of 5-methyl group to a pyrimidine base is detrimental to 3'-dNTP's potency. Direct evidence was obtained that all four canonical 3'-dNTP are incorporated into elongating RNA chains and the incorporation terminates NS5B RdRp-catalyzed RNA synthesis. The K(i) values for each of 3'-dNTPs were determined in the single nucleotide incorporation experiments. The nucleoside form of 3'-dNTPs was further evaluated in a cell culture-based HCV subgenomic replicon assay. The discrepancy between the potent in vitro activity and the weak cellular activity of these chain terminators was discussed in the context of nucleoside metabolism. This proof of concept study demonstrates that canonical 3'-dNTPs can function as an effective chain terminator for HCV NS5B RdRp with cytidine as the preferred nucleoside scaffold. Our results further sheds light on the potential hurdles that need to be overcome for successful development of active nucleoside chain terminators in vivo for a viral RNA polymerase, especially the HCV NS5B RdRp. PMID- 12767473 TI - Aspirin-like molecules that inhibit human immunodeficiency virus 1 replication. AB - Some anti-inflammatory molecules are also known to possess anti-human immunodeficiency virus (HIV) activity. We found that o-(acetoxyphenyl)hept-2-ynyl sulfide (APHS), a recently synthesized non-steroidal anti-inflammatory molecule can inhibit HIV-1 replication. The aim of this study was to clarify the mechanism of action of APHS. When administered during the first steps of the infection, APHS was capable of inhibiting the replication of several HIV-1 strains (macrophage-tropic and/or lymphocytotropic) in a dose-dependent manner in both peripheral blood mononuclear cells (PBMC), monocyte-derived macrophages and peripheral blood lymphocytes with 50% inhibitory concentration values of approximately 10 microM. The 50% toxic concentration of APHS varied between 100 and 200 microM in the different primary cells tested. APHS did not affect HIV-1 replication once the provirus was already inserted into the cellular genome. APHS also did not inhibit HIV-1 entry into the host cells as determined by quantification of gag RNA inside PBMC 2h after infection. However, APHS did inhibit gag DNA synthesis during reverse transcription in primary cells, which indicates that APHS may target the reverse transcription process. PMID- 12767474 TI - Broad spectrum anti-RNA virus activities of titanium and vanadium substituted polyoxotungstates. AB - Seven polyoxotungstates substituted with vanadium or titanium atoms were examined for their activity against Flaviviridae (Dengue fever virus, DFV), Orthomyxoviridae (influenza virus type A, fluV-A), Paramyxoviridae (respiratory syncytial virus, RSV, parainfluenza virus type 2, PfluV-2 and canine distemper virus, CDV) and Lentiviridae (human immunodeficiency virus type 1, HIV-1) families. Among the seven polyoxotungstates examined, PM-43 [K(5)[SiVW(11)O(40)]], PM-47 [K(7)[BVW(11)O(40)]], and PM-1001 [K(10)Na(VO)(3)(SbW(9)O(33))(2)]26H(2)O contained vanadium. PM-1002 had the same core structure of (VO)(3)(SbW(9)O(33))(2) as PM-1001; however, three V atoms of PM-1001 consisted of two V(IV) and one V(V) and those of PM-1002 consisted of three V(IV). On the other hand, PM-518 [[Et(2)NH(2)](7)[PTi(2)W(10)O(40)]], PM 520 [Pri(2)NH(2)](5)[PTiW(11)O(40)] and PM-523 [PriNH(3)](6)H[PTi(2)W(10)O(38)(O(2))(2)]H(2)O all contained titanium. All compounds showed broad spectrum antiviral activity against all viruses examined except for PMs-43, -518 and -523 which did not exhibit inhibitory activity at >/=50 microM against PfluV-2, CDV and DFV, respectively. All compounds were inhibitory against HIV replication at an EC(50) of less than 2.0 microM. Among them, PMs-1001 and -1002 showed the most potent inhibition. The compounds were not toxic for MDCK, HEp-2 and Vero cells at a concentration of 200 microM. For the exponentially growing MT-4 cells, the vanadium containing polyoxometalates (PMs-43, 47, 1001, 1002) showed toxicity at concentrations between 41 and 47 microM. On the other hand, titanium containing polyoxometalates (PMs-518, -520, 523) were not toxic at 100 microM. The mechanism of anti-HIV action of PM-1001 was analyzed: it affected the binding of HIV to the cell membrane and syncytium formation between HIV-infected and uninfected cells. PMID- 12767475 TI - AMPA receptor trafficking and synaptic plasticity: major unanswered questions. AB - Dynamic movements of AMPA receptors in and out of the postsynaptic membrane account for, at least in part, the expression of NMDA receptor-dependent changes in synaptic efficacy such as long-term potentiation and long-term depression. Recently some of key molecules and subunit rules involved in AMPA receptor trafficking have been identified. In this update article, we try to highlight what we believe to be the major conceptual problems and unanswered questions in this rapidly moving field of neuroscience. PMID- 12767476 TI - Regenerative process of the facial nerve: rate of regeneration of fibers and their bifurcations. AB - After the main trunk of the mouse facial nerve was injured by crushing, a fiber tracing method was used to quantify the facial motor neurons that extended regenerating nerve fibers to the specific site of the facial nerve branch. The total number of motor neurons retrogradely labeled with a fluorescent tracer, Fluoro-Gold (FG), were 0 on postsurgical days (PSDs) 1 and 2, 75+/-25 on PSD3, 264+/-21 on PSD4, 378+/-19 on PSD6, 428+/-19 on PSD8, 491+/-13 on PSD12 and 532+/ 15 on PSD16. Assuming that the FG-positive neurons (535+/-11) of the control mice represent 100%, the FG-labeled neurons accounted for 0, 14, 49, 71, 80, 92 and 99% on the corresponding days. Two different fluorescent tracers were applied to the different facial nerve branches 16 days after facial nerve injuries. Double labeled neurons were consistently found in the nerve-crushed facial nucleus (3.2%), and their number increased in the nerve-transected facial nucleus (12.2%). The present study indicates that the regenerating facial nerve consists of heterogeneous nerve fibers with varying growth rates and that excessive axonal branching occurs more frequently in the nerve-transected than in the nerve crushed injuries. PMID- 12767477 TI - Time course of tonal frequency-response-area of primary auditory cortex neurons in alert cats. AB - Cells in the A1 auditory cortex of alert animals show various response time courses during pure-tone stimuli: tonic, phasic-tonic, and phasic. Previously the time course of the spike firing rates was examined at a characteristic frequency (CF) or in a range of frequencies including CF. We investigated time-course of the frequency-response-area (FRA) during pure-tone stimuli in A1 cells of alert cats. The short rise/fall time (0.1-2 ms) and long stimulus duration (0.5 s) was used for investigation of the time course. FRA changed with time drastically in the phasic cells, mildly in the phasic-tonic cells, but not in the tonic cells. The best-response frequency (BF) within FRA was constant throughout the stimulus duration in the tonic and phasic-tonic cells, but was difficult to define in the phasic cells. The phasic firing properties of the phasic cells were preserved even during the bandnoise stimuli at various bandwidth and spectral locations. The variability of FRA time-course between cell types may play a role for analyzing auditory spectral cues that vary with a wide range of time constant. PMID- 12767478 TI - The effects of dentate granule cell destruction on behavioral activity and Fos protein expression induced by systemic MDMA in rats. AB - In this study, we examined the effect of the s.c. administration of (+/-) 3,4 methylenedioxymethamphetamine (MDMA) or saline on locomotor activity and Fos expression following the bilateral destruction of hippocampal dentate granule cells by colchicine in rats. The lesioned animals, when administered s.c. saline, showed a significantly greater increase in locomotor activity compared to the intact animals, and revealed a marginally significant level of increased locomotor activity compared to the sham-lesioned animals. In addition, when the lesioned animals were given s.c. saline or MDMA, there was a significant increase in Fos expression in the nucleus accumbens core, but not in the medial prefrontal cortex, dorsolateral prefrontal cortex, anterior cingulate cortex, piriform cortex, dorsal striatum, or nucleus accumbens shell, compared to the intact and sham-lesioned animals. Overall, these results suggest that the nucleus accumbens core may be involved in the enhancement of locomotor activity induced by the injection of saline alone (stress loading) or MDMA following bilateral destruction of hippocampal dentate granule cells by colchicine. PMID- 12767479 TI - Neurites undergoing Wallerian degeneration show an apoptotic-like process with Annexin V positive staining and loss of mitochondrial membrane potential. AB - Wallerian degeneration, the disintegration of the distal part of an injured axon, is an important event in many neurodegenerative diseases. We studied Wallerian degeneration in dorsal root ganglion (DRG) explants in culture by separating neurites from their cell bodies with a scalpel. The severed neurites showed Annexin V positive staining, that spreads distally with a rate comparable to that of slow axonal transport in intact neurons in vivo. Moreover, the injured neurites showed loss of mitochondrial membrane potential. These features resemble those seen when cells undergo apoptosis. These data contribute to a new understanding of the mechanism of axonal degeneration, have implications for the response of stromal cells in central nervous system (CNS) and raise the prospect of new pharmacological treatments for those neurodegenerative pathologies where the protection of the cell body alone does not alleviate the disease. PMID- 12767480 TI - Anisotropic functional connections between the auditory cortex and area 18a in rat cerebral slices. AB - We developed a new method to visualize the myeloarchitecture in fresh slices, and investigated the properties of the functional neural connections around the boundary between the primary auditory cortex (area 41) and area 18a in rat cerebral slices. A fresh slice illuminated by near-vertical light was observed with a CCD camera. The translucent images of the slice showed contrast patterns very similar to myeloarchitecture. The boundary between these areas was identified by the well-developed layer IV/V in area 41 but not in area 18a. Antidromic/presynaptic components of the field potentials stimulated and recorded across the areal boundary showed symmetric distribution, while the postsynaptic field potentials in the direction from area 41 to 18a were more prominent than those in the opposite direction in layer II/III. In contrast, the dominant direction of propagation of postsynaptic potentials was from area 18a to 41 in layer V. In the presence of 1 microM bicuculline, an inhibitor of GABA(A) receptors, the polysynaptic activities propagating from area 18a into 41 via layer V were elicited by stimulation of area 18a. The propagation measured by Ca(2+) imaging or field potential recordings was potentiated after both areas 18a and 41 were alternately stimulated several times. PMID- 12767481 TI - The nucleus accumbens unit activities related to the emotional significance of complex environmental stimuli in freely moving cats. AB - To establish the functional role of the nucleus accumbens (ACC) in emotionally motivated behaviors, extracellular single or multiple units were recorded from the ACC under an unanesthetized, freely moving non-operant condition in 43 cats. Then neuronal and behavioral responses to stimuli such as tones, live small animals, a human, air puffs and so on were analyzed using a videotape monitoring method. A total of 98 units were recorded from the ACC, and 34 (34.7%) of them responded to some of the stimuli. Of the 34 responded units, 5 (15%) units responded to quiet approach-provoking stimuli (appetitive stimuli), and 29 (85%) units responded to escape- or defensive attack-provoking stimuli (aversive stimuli). Although most of the units responded in an individual item-specific manner, there was no unit reacted to both the appetitive and aversive stimuli. The neuronal responses were not related to any movement or locomotion. The results were compared with those with the amygdala, and their neuronal responsiveness were shown to be quite different from each other. Although the ACC appears to be involved in the central processing of emotional behavior in some way, the role was suggested to be entirely different from that of the amygdala. PMID- 12767482 TI - Antioxidant NAC and AMPA/KA receptor antagonist DNQX inhibited JNK3 activation following global ischemia in rat hippocampus. AB - c-Jun N-terminal kinase-3 (JNK3), the only neural-specific isoform, may play an important role in excitotoxicity and neuronal injury. To analyze the variation of JNK3 activation, levels of phospho-JNK3 were measured at various time points of ischemia and selected time points of reperfusion, respectively. Our study illustrated that JNK3 was rapidly activated and translocated from cytosol to nucleus during ischemia. During reperfusion, two peaks of JNK3 activation occurred at 30 min and 3 days, respectively. To further define the mechanism of JNK3 activation, antioxidant N-acetylcysteine (NAC), alpha-amino-3-hydroxyl-5 methyl-4-isoxazolepropionic acid (AMPA)/kainate (KA) receptor antagonist 6,7 dinitro-quinoxaline-2,3(1H,4H)-dione (DNQX), N-methyl-D-aspartate (NMDA) receptor antagonist ketamine and L-type voltage-gated Ca(2+) channel (L-VGCC) antagonist nifedipine were given to the rats 20 min prior to ischemia. The results showed that NAC obviously inhibited JNK3 activation during the early reperfusion, whereas DNQX preferably attenuated JNK3 activation during the latter reperfusion. Ketamine and nifedipine had no significant effects on JNK3 activation during reperfusion. Consequently, reactive oxygen species (ROS) and AMPA/KA receptor were closely associated with JNK3 activation following global ischemia. PMID- 12767483 TI - Unaltered pain-related behavior in mice lacking NMDA receptor GluRepsilon 1 subunit. AB - Noxious afferent input following tissue damage and inflammation triggers a state of neuronal hyperexcitability-a phenomenon of central sensitization-which manifests behaviorally as allodynia and hyperalgesia. At the molecular level, maintenance of central sensitization is largely dependent on the N-methyl-D aspartate receptor (NMDAR) activation. NMDARs are composed of GluRzeta1 (NR1) and one of four GluRepsilon (NR2) subunits, which determine the functional properties of native NMDARs. Although there is accumulating evidence to implicate GluRepsilon 2-containing NMDARs in pain mechanisms, the functional significance of GluRepsilon 1-containing NMDARs in this setting has not been examined in detail. Here, we used hind paw injection of formalin, complete Freund's adjuvant and a nerve injury model to investigate the effects of GluRepsilon 1 subunit gene deletion on pain-related behavior in mice. In all of the models tested, GluRepsilon 1-deficient mice exhibited responses similar to wild-type controls. These results suggest that GluRepsilon 1 disruption does not result in altered nociceptive behavior in mice. Although the contribution of other nociceptive pathways cannot be ruled out, we speculate that the preserved function of GluRepsilon 2-containing NMDARs could explain unaltered nociceptive behavior in mutant mice. PMID- 12767484 TI - Gonadal and adrenal effects on the glucocorticoid receptor in the rat hippocampus, with special reference to regulation by estrogen from an immunohistochemical view-point. AB - Focusing on the hippocampal CA1 region, effects of peripheral gonadal and adrenal steroids on the glucocorticoid receptor (GR) were immunohistochemically evaluated in male and female adult rat brains after adrenalectomy (ADX), gonadectomy (GDX), and administration of estradiol (E2) and/or corticosterone (CS). In ADXed male rats, the hippocampal nuclear GR decreased and turned back to the cytoplasm, whereas in females, nuclear localization persisted even after ADX. In GDX+ADXed female rats, the GR was dispersedly translocated from the nucleus to the cytoplasm as well as in GDX+ADXed males. The dispersed cytoplasmic GR was again translocated into the nucleus by administration of CS. In addition, administration of a small dose of E2 for 4-13 days was found to sufficiently recover the nuclear location of GR in GDX+ADXed rat brains, whereas medium-to large doses could not do this. Also, a longer administration more strongly enhances the nuclear GR location and expression. The present study provided strong immunohistochemical evidence that the sexually dimorphic effects of ADX on hippocampal GR are attributable to gonadal hormones, and that E2 is implicated in the effects in inversely-dose- and directly-duration-dependent manner. Taken together, intriguing gonadal and adrenal crosstalk is considered to play some important role in regulating hippocampal GR morphology and to have a possibly crucial influence on stress-related disorders such as depression. PMID- 12767485 TI - Reproducibility of callosal effects of transcranial magnetic stimulation (TMS) with interhemispheric paired pulses. AB - Transcranial magnetic stimulation (TMS) of the motor cortex of one hemisphere (conditioning stimulus (CS)) inhibits EMG responses evoked in distal hand muscles by a later magnetic stimulus given at an appropriate interval, over the opposite hemisphere (test stimulus (TS)). This effect is commonly attributed to an inhibition produced at cortical level via a transcallosal route. The present study assessed the reproducibility of the transcallosal inhibition effects in different sessions in healthy subjects. Within- and between-subject variability, relating to interhemispheric differences was also evaluated. A magnetic CS on one hemisphere effectively inhibited EMG responses of the abductor digiti minimi stimulated by a TS delivered over the opposite hemisphere in a range of intervals centered at 12 ms. Even though group effects were reproduced in separate sessions, the high between- and within-subject variability yielded low test retest correlations. This differentiation forces the definition of reproducibility (or repeatability), as the replication of the same mean curves of EMG reduction, and of reliability, as the between- or within-subject correlations between values of specific EMG measures. PMID- 12767486 TI - Rhythmic phrenic, intercostal and sympathetic activity in relation to limb and trunk motor activity in spinal cats. AB - During L-DOPA-induced fictive spinal locomotion rhythmic activities in nerves to internal intercostal and external oblique abdominal muscles and in phrenic and sympathetic nerves were observed which were always coordinated with locomotor activity in forelimb and hindlimb muscle nerves. A periodicity with longer lasting tonic phases could be induced by cutaneous nerve stimulation or asphyxia. This activity was observed in limb motor nerves as well as in respiratory motor and sympathetic nerves. A slow independent activity of the phrenic and intercostal nerves or the sympathetic nerves, which could be related to a normal respiratory rhythm or independent sympathetic rhythms was not observed. The findings indicate that during fictive spinal locomotion the activity of spinal rhythm generators for locomotion also projects onto respiratory and sympathetic spinal neurones. PMID- 12767487 TI - Astrocyte-derived factors instruct differentiation of embryonic stem cells into neurons. AB - Pluripotent embryonic stem (ES) cells may differentiate into neurons in vitro. This is valuable in the study of neurogenesis and in the generation of donor cells for neuronal transplantation. Here we show that astrocyte-derived factors instruct mouse and primate ES cells to differentiate into neurons. Cultured in astrocyte-conditioned medium (ACM) under free-floating conditions, within 4 days, colonies of undifferentiated mouse ES cells give rise to floating spheres of concentric stratiform structure with a periphery of neural stem cells, which are termed Neural Stem Spheres. Culturing the spheres on an adhesive substrate in ACM promotes neurogenesis, and cells in the spheres differentiate into neurons within 5 days, including dopaminergic neurons. In contrast, neither astrocytes nor oligodendrocytes are formed. The procedure developed for mouse ES cells can be applied to monkey ES cells. This neurogenesis pathway provides a new insight into mechanisms of specification of cell fates in early development and also provides a simple procedure for fast and efficient generation of a vast number of neural stem cells and neurons. PMID- 12767488 TI - Insensitivity of the microdialysis zero-net-flux method to nonlinear uptake and release processes. AB - In the microdialysis zero-net-flux (ZNF) method the extraction efficiency is conventionally obtained by linear regression. The linear analysis may become invalid for certain analytes that have nonlinear uptake/release processes in the tissue. To examine this hypothesis, a nonlinear model was used to numerically investigate the nonlinearity of the ZNF plot caused by nonlinear uptake and release processes. Three findings from this analysis are: (i) the ZNF method is markedly insensitive to the nonlinear active processes; (ii) a slow infusion rate or a long probe membrane can suppress the nonlinearity; (iii) the release under autoreceptor control does not affect the slope and linearity of the concentration difference plot. It is concluded that in the nM infusion range, the ZNF method is unable to distinguish whether or not the tissue clearance process is nonlinear. During electrical stimulation, neurotransmitter overflow may cause the microdialysis ZNF method to exhibit nonlinearity. PMID- 12767489 TI - Transcranial magnetic stimulation over the cerebellum evokes late potential in the soleus muscle. AB - Transcranial magnetic stimulation (TMS) with a double cone coil placed over the left lateral side of the basal occiput was able to elicit late electromyographic (EMG) responses at the bilateral soleus muscles (SOL) averaged over 30 stimulation events, with a mean latency of approximately 100 ms. These EMG responses were detected using a low frequency bandpass filter with 0.05 Hz magnetic stimulation on ten healthy subjects in standing posture. As magnetic stimulation over the left basal occiput with a double cone coil can stimulate cerebellar structure, this late response seems to be conducted from the cerebellar structure to the SOL via an as yet unknown descending pathway. Here, we report new late EMG responses in relation to cerebellum or cerebellum related structures. PMID- 12767490 TI - MRI cytoarchitectonics: the next level? PMID- 12767491 TI - Effects of repetitive transcranial magnetic stimulation on behavioral and neurochemical changes in rats during an elevated plus-maze test. AB - In transcranial magnetic stimulation (TMS) the regional electrical activity in the brain is influenced by a pulsed magnetic field. The rapidly changed magnetic field produces electrical currents that activate neurons. Repetitive TMS (rTMS) treatment can cause functional changes in the cortex. The present study clarified the effects of rTMS treatment on behavioral changes in rats, focusing on anxiety by using an elevated plus-maze (plus-maze) test. The effects of rTMS treatment on neurochemical changes during the plus-maze test were investigated by determining the extracellular levels of serotonin (5-HT) and dopamine (DA) in the prefrontal cortex by using in vivo microdialysis. Each rat received rTMS of the frontal brain for 3 days, during which 125 stimuli from five trains in a day were applied at 25 Hz for 1 s with 2-min intervals between trains. Three-day series of (chronic) rTMS treatment caused significant increases in the time spent in open arms and the number of entries into open arms of the plus-maze compared with non treated and sham-treated rats, which were not observed in 1-day series of (acute) rTMS treatment. Chronic rTMS treatment suppressed the increases in 5-HT levels induced by the plus-maze test, but did not influence the elicited DA levels. These data suggest that chronic rTMS treatment of the frontal brain has anxiolytic effects in rats, which are related to the 5-HTergic neuronal system. PMID- 12767492 TI - Brain aging in normal Egyptians: cognition, education, personality, genetic and immunological study. AB - Studying the cognitive and immunological changes that occur in old age as well as genetic function have been considered an important subject to differentiate between normal brain aging and early dementia especially Alzheimer's disease. The aim of this study is to stress on age-related neuropsychological and electrophysiological (P(300)) changes in normal Egyptian subjects, to throw light on the value of genetic (Apo-E(4) genotype) and immunological markers [interleukin-6 (IL-6) and intercellular adhesion molecules (ICAM-1) in the serum] as tools used in early detection of cognitive decline in cerebral aging. Ninety four normal Egyptian subjects (below and above 60 years) were submitted to the following: (1) neuropsychological tests for testing memory, perception, psychomotor performance and attention, (2) Eysenck Personality Questionnaire (EPQ) for personality traits, (3) event-related potential study (P(300), latency and amplitude), (4) genetic test for detection of Apolipoprotein E genotype and (5) immunological studies including detection of the level of IL-6 and ICAM-1 in serum. There was a significant impairment of memory, psychomotor performance and perception in elderly subjects particularly males and subjects with low level of education. Regarding personality, significantly high scores were obtained in neuroticism scale of EPQ in elderly subjects. Apo-E(3)/E(3) was the most common genotype encountered in Egyptian subjects (49.1%). It was found that subjects with Apo-E(4) genotype did significantly worse in scores of intentional memory test (sensory memory) when compared with other genotypes. Statistically significant impairment in attention and sensory memory was found in subjects with high IL-6 level. This could not be detected in subjects with high ICAM-1 level. In conclusion, advancing age and lower levels of education are considered risk factors for cognitive decline in normal brain aging. Neuropsychological tests remain as the highly sensitive tools for detection of early cognitive impairment. Neurotic traits are more encountered in old age. Apo-E(4) genotype is associated with significant sensory (intentional) memory impairment. High IL-6 level in the serum is accompanied by significant impairment in attention and sensory (intentional) memory. PMID- 12767493 TI - Protein and gene analyses of dysferlinopathy in a large group of Japanese muscular dystrophy patients. AB - Mutations in the dysferlin gene cause muscular dystrophies called dysferlinopathy, which include limb-girdle muscular dystrophy type 2B (LGMD2B) and Miyoshi myopathy (MM). To clarify the frequency, clinicopathological and genetic features of dysferlinopathy in Japan, we performed protein and gene analyses of dysferlin. We examined a total of 107 unrelated Japanese patients, including 53 unclassified LGMD, 28 MM and 26 other neuromuscular disorders (ONMD). Expression of dysferlin protein was observed using immunohistochemistry (IHC) and mini-multiplex Western blotting (MMW), and mutation analysis was performed. We found a deficiency of dysferlin protein by both IHC and MMW in 19% of LGMD and 75% of MM patients, and mutations in the dysferlin gene were identified in this group alone. 19% of dysferlin-deficient patients had 3370G-->T missense mutation and 16% had 1939C-->G nonsense mutation. The patients with homozygous 3370G-->T mutation showed milder clinical phenotypes. Twenty-five percent of MM muscles had normal dysferlin protein contents that suggested the genetic heterogeneity of this disease. Altered immunolocalization of dysferlin was observed in not only primary dysferlinopathy, but also in the several diseased muscles with normal protein contents. This result implies the necessity of other protein(s) for proper membrane localization of dysferlin, or some roles of dysferlin in the cytoplasmic region. PMID- 12767494 TI - Developmental stages of cortical Lewy bodies and their relation to axonal transport blockage in brains of patients with dementia with Lewy bodies. AB - We investigated 10 dementia with Lewy bodies (DLB) brains showing various degrees of Lewy pathology using alpha-synuclein-immunohistochemistry, and morphologically divided cortical Lewy bodies (LB) into six developmental stages. Further, we demonstrated the ultrastructure of each stage of cortical LB using alpha synuclein-immunoelectron microscopy. In the initial stage, alpha-synuclein accumulated in part of the neuronal cytoplasm without filamentous components, then formed LB and LB-related neurites composed of granulo-filamentous components. Finally, LB degraded to extracellular LB composed of loose filamentous components with involved astroglial processes. In addition, we immunohistochemically investigated the accumulation of axonal transported substances in cortical LB, and showed that APP, chromogranin-A, synphilin-1 and synaptophysin accumulated in cortical LB from stages 1, 2, 3 and 4, respectively. These findings suggest that chronic axonal transport blockage is implicated in the development of cortical LB in DLB brains. PMID- 12767495 TI - Treatment of status epilepticus: a survey of neurologists. AB - BACKGROUND: New antiepileptic drugs (AEDs) have provided alternatives to traditional treatment paradigms for status epilepticus (SE). METHODS: To determine current treatment preferences for generalized convulsive status epilepticus (GCSE), we surveyed 106 members of the Critical Care or Epilepsy sections of the American Academy of Neurology. RESULTS: Most respondents initially treat patients with intravenous (IV) lorazepam (76%), followed by phenytoin or fosphenytoin (95%) if first-line therapy fails. Preferences for GCSE refractory to two AEDs (RSE) varied: 43% would give phenobarbital, 19% would give one of three continuous-infusion (cIV) AEDs (pentobarbital, midazolam, propofol), and 16% would give IV valproic acid. About half indicated "burst suppression" (56%) and half indicated "elimination of seizures" (41%) as the titration goal for cIV-AED therapy. About half (42%) would add a new cIV-AED, and the other half (41%) would not add another agent to treat electrographic SE refractory to four AEDs. DISCUSSION: In accordance with published trials and general guidelines, neurologists most often use lorazepam followed by phenytoin or fosphenytoin as first-line and second-line therapies for GCSE. There is no consensus for third line or fourth-line treatment for RSE. The treatment of RSE needs to be studied in a large, prospective, randomized, multicenter trial. PMID- 12767496 TI - Long-lasting effect of transcutaneous electrical nerve stimulation on the thermal hyperalgesia in the rat model of peripheral neuropathy. AB - We demonstrate here unexpectedly long-lasting effect of transcutaneous electrical nerve stimulation (TENS) to alleviate thermal hyperalgesia in rats with peripheral neuropathy produced by constriction of sciatic nerve. For TENS groups, electrical stimulation for 16.7 min (1 Hz, paired current, 12 mA, 5-ms interval, 0.2-ms duration, 999 pairs), once a day, was delivered for 5 consecutive days, under halothane anesthesia (Hal-TENS group) or pentobarbital anesthesia (Pent TENS group). For non-TENS groups, only the anesthesia was delivered (Hal-no TENS group, Pent-no TENS group). For the control group, neither anesthetics nor TENS was delivered. To evaluate hyperalgesia, paw withdrawal latency (PWL) to radiant heat was measured before nerve constriction and five times after the constriction; just before TENS and at 1, 3, 7, and 14 days after the completion of TENS. Compared to the non-TENS groups, rats in the TENS groups showed significantly reduced thermal hyperalgesia at least for 3 days (Pent-TENS group) or for 7 days (Hal-TENS group) after TENS. These results indicate a possible long lasting therapeutic effect of TENS applied under general anesthesia. PMID- 12767497 TI - The prevalence and clinical characteristics of MS in northern Japan. AB - In Japan, there is a low prevalence rate (PR) of multiple sclerosis (MS; 0.8 4.0/100,000) but a relatively high frequency of "optic-spinal form" MS (OS-MS). There have been no intensive epidemiologic frequency studies, however, in over 30 years. We performed a province-wide prevalence study of MS in the Tokachi province of Hokkaido, the northernmost island of Japan, and compared the observed clinical features with other populations in Japan and Western countries. Prevalence was determined on March 31, 2001. The primary sources for the case ascertainment were 13 hospitals that treated patients with neurologic diseases including MS in Tokachi. Patients were classified according to Poser's criteria. The prevalence rate of clinically definite or laboratory-supported definite MS (LSDMS) was 8.57 per 100,000 [31/361,726; male/female ratio=1:2.9, and age at onset=29.1+/-14.2 (mean+/-SD) years]. Out of the 31 patients, 5 (16%) were classified as OS-MS. The prevalence rate of MS in the Tokachi province was the highest reported in Orientals to date, although still low in comparison with Western communities at a similar latitude. In contrast to the previous reports in Japan, there was a relatively low frequency of OS-MS in Hokkaido. PMID- 12767498 TI - Health-related quality of life and depression in an Italian sample of multiple sclerosis patients. AB - Only few publications have been reported on Health-related Quality of Life (HRQoL) in patients with multiple sclerosis (MS). EDSS is the most common outcome measure for either impairment or disability of MS, but it is not able to catch other aspects of MS impact on HRQoL. The authors performed a cross-sectional study on the group of all patients with MS who were diagnosed at least 4 years before 1998 in Catania (South Italy). One hundred and eighty patients out of 308 were enrolled in the study. SF-36 was used to catch the HRQoL of MS patients. EDSS, Beck Depression Inventory (BDI) and time since diagnosis were investigated as variables affecting the HRQoL of MS patients. The patients showed significant lower mean scores for all SF-36 health dimensions compared with sex- and age adjusted scores in a general healthy Italian population (p<0.001). EDSS scores correlated only with physical functioning (r=-0.76 p<0.001). As expected, the more severe was the disease, the longer its duration and the lower the patients' skillness on HRQoL. BDI showed high partial correlations with all SF-36 health domains with r=-0.38 to -0.65 (p<0.001). This study showed that SF-36 is able to assess the HRQoL of MS patients. Depression strongly influenced the HRQoL of MS patients. EDSS and time since diagnosis also affected the HRQoL of MS patients. Our results are comparable with other European studies. PMID- 12767499 TI - Clinical progression of mitochondrial myopathy is associated with the random accumulation of cytochrome c oxidase negative skeletal muscle fibres. AB - We studied the accumulation of cytochrome c oxidase (COX)-negative skeletal muscle fibres in six patients with a myopathy due to a mitochondrial DNA (mtDNA) defect. Each patient was biopsied on two or more occasions over a period of 3-15 years. Progressive proximal weakness was associated with an increase in the proportion of COX-negative fibres. These fibres were arranged randomly, indicating that each fibre became COX negative independently of the status of neighbouring fibres. The clinical progression of mtDNA myopathy is therefore a consequence of a biochemical defect that develops independently within individual muscle fibres. It is likely that this is due to the clonal expansion of mutant mtDNA. PMID- 12767500 TI - Antibodies against myelin oligodendrocyte glycoprotein in the cerebrospinal fluid of multiple sclerosis patients. AB - Antibodies against myelin oligodendrocyte glycoprotein (MOG) mediate demyelination in experimental autoimmune encephalomyelitis (EAE) in different animal species and are implicated in the immunopathogenesis of multiple sclerosis (MS). In order to evaluate the anti-MOG response, we have analyzed the cerebrospinal fluids (CSFs) from 44 MS patients and 51 controls, 11 with other inflammatory neurological disorders (OIND) and 40 with non-inflammatory neurological disorders (NIND). The frequency of anti-MOG antibodies positive patients in the MS group (30%) was significantly higher compared to the NIND (8%, p=0.02), but not compared to the OIND group (55%, p=0.228). Interestingly, all six patients with neurosarcoidosis had MOG-specific antibodies in their CSF. Frequency of anti-MOG antibodies was similar in patients with clinically active and stable MS (32% and 26%, respectively; p=0.921). However, in clinically active MS patients, antibody titers were higher in comparison with patients with stable disease, although the difference did not reach the level of statistical significance (p=0.06). These results further support the potential role of anti MOG antibodies in the immunopathology of MS in the subset of patients with this disease. Furthermore, our findings suggest for the first time that anti-MOG antibodies could be an accessory diagnostic tool in neurosarcoidosis. PMID- 12767501 TI - Magnetic resonance imaging identifies cytoarchitectonic subtypes of the normal human cerebral cortex. AB - BACKGROUND: Magnetic Resonance Imaging (MRI) allows a detailed "in vivo" macroscopic study of the human brain; previously, it has been demonstrated that Fluid Attenuated Inversion Recovery (FLAIR) sequence shows higher signal intensity of cortices belonging to limbic structures. PURPOSE: To measure and compare signal intensities (SI) of cytoarchitectonically different cortical regions. METHODS: In 22 adult subjects, without psychiatric or neurological diseases, FLAIR sequence was performed in coronal slices, perpendicular to the main hippocampal axis. Signal intensity was measured, with a region-of-interest (ROI) function, in 12 different cortical regions. We compared these values and grouped the cortices into five groups: (1) limbic cortices, (2) paralimbic agranular cortices, (3) paralimbic granular cortices, (4) parietal-type neopallium, (5) frontal-type neopallium. A t-test for comparison of paired samples was performed, considering pA polymorphism in the promoter region of E-cadherin has been shown to decrease gene transcription. This allelic variation may be a potential genetic marker that can help identify those individuals at higher risk for invasive/metastatic disease. We studied the effect of E-cadherin gene polymorphism on urothelial cancer susceptibility in a case control study of 314 urothelial cancer patients and 314 age-sex matched controls, to determine whether this polymorphism is a biomarker for the risk and how aggressive the disease is. The frequency with which the subjects carried E cadherin A/A genotype was significantly higher in the urothelial cancer patients than in the healthy control subjects (OR=2.32, 95% CI 1.03-5.22). Subdividing urothelial cancer according to tumor differentiation and stage, we found no association between E-cadherin polymorphism and poorly-differentiation and invasiveness of urothelial cancer. Furthermore, no significant association between E-cadherin polymorphism and recurrence rate of urothelial cancer patients was found. The present study demonstrates for the first time that E-cadherin A/A genotype may be associated with susceptibility to urothelial cancer, but not with the progression of disease. PMID- 12767512 TI - Allelic loss of DNA locus of the RET proto-oncogene in small cell lung cancer. AB - We analyzed all 21 exons of the RET proto-oncogene of paired genomic DNA from tumors and normal tissues in 12 small cell lung cancer (SCLC) patients for the presence of genetic alteration. Polymerase chain reaction single-strand conformation polymorphism analysis and direct sequencing revealed that heterozygosity of the RET proto-oncogene was lost in the tumor tissues of six patients out of eight informative SCLC patients, although point mutation was not evident in any tumors. These results suggest that a deletion of the chromosomal region including the RET proto-oncogene is involved in the pathogenesis of SCLC. PMID- 12767513 TI - Connexin 37 gene is not mutated in lung carcinomas 3LL and CMT. AB - Our previous studies have shown that intercellular communication mediated by gap junctions is impaired in most tumors as well as in cancer cell lines. However, connexin genes that encode gap junction proteins are only rarely mutated in cancer cells. On the other hand, it was reported that mutated Connexin 37 (Cx37) is the origin of shared tumor-associated antigenic octa-peptides (MUT 1 and MUT 2) of two independently derived lung carcinomas 3LL and CMT 64 of mouse origin. Two Cx37 mutations have been implicated: a Cys-54-Gln substitution in FEQNTAQP (MUT 1) and FEQNTAQA (MUT 2); an additional Pro-59-Ala substitution has been proposed in MUT 2. A Cys-54-Gln mutation in both tumors requires three base changes (TGT-to-CAG) to have occurred twice in independently derived tumors. Another complication stems from the fact that Cys 54, which is located in the extra-cellular domain is conserved in all connexins. Due to the important implications that these findings may have regarding the role of gap junctional communication in lung carcinomas as well as in the origin of tumor-associated antigens, we decided to re-examine these mutations. Thus, we PCR-amplified genomic DNA from 3LL and CMT and sequenced the coding region of Cx37 encompassing codon 54. We then analyzed the PCR products by digestion with the restriction enzyme MaeIII, to discern the presence of the putative mutation. Here we have unambiguously demonstrated that clones K(b)39.5 (39.5) and D122 of 3LL, and C6 and E9 of CMT 64, previously employed, have only normal Cx37 sequences, including those of codon 54. Therefore, we concluded that Cx37 is not mutated in 3LL and CMT 64 carcinomas. PMID- 12767514 TI - pH effects on the cellular uptake of four photosensitizing drugs evaluated for use in photodynamic therapy of cancer. AB - The difference in extracellular pH in malignant as compared to normal healthy tissues has been proposed to contribute to selective uptake of photosensitizers in tumors. Hematoporphyrin IX (HpIX), disulfonated meso-tetraphenylporphine (TPPS(2a)), meso-tetra(3-hydroxyphenyl)porphine (mTHPP) and meso-tetra(3 hydroxyphenyl)chlorin (mTHPC) were chosen to examine the pH dependence of their cellular drug uptake. The study was performed in the pH range 6.5-8.0 and showed that significantly higher amounts of the drug are taken up by T-47D cells at low pH values only in the case of HpIX. The pH value of the incubation medium did not influence the cellular uptake of mTHPP, mTHPC and TPPS(2a) significantly. The present work indicates that tumor selectivity of dyes, which get more lipophilic with decreasing pH value, may be related to the low extracellular pH value. PMID- 12767515 TI - Association of CYP17 genetic polymorphism with intra-tumoral estradiol concentrations but not with CYP17 messenger RNA levels in breast cancer tissue. AB - The variant allele(1931C) of CYP17 (1931C/T), which is one of the key enzymes involved in estrogens synthesis, has been shown to be associated with breast cancer risk. Since this variant allele creates an additional putative Sp-1 binding site (CCACC) in the promoter region, it is speculated that it enhances the transcription of CYP17, leading to the enhanced estrogens synthesis in breast tumors. CYP17 messenger RNA (mRNA) expression could be detected in all the normal breast (n=51) and tumor tissues (n=67) by a real-time polymerase chain reaction but CYP17 mRNA expression was not significantly different between the variant allele carriers and non-carriers. In addition, no significant correlation was observed between CYP17 mRNA and E2 levels in tumors, indicating an unimportant role of CYP17 in in situ synthesis of E2. On the other hand, intra-tumoral E2 levels were significantly (P=0.025) higher in the variant allele carriers (127.2+/-11.0 pg/g) than non-carriers (88.2+/-8.5 pg/g). Since it has been previously reported that serum E2 levels are higher in variant allele carriers than non-carriers, it is speculated that the higher intra-tumoral E2 levels in the variant allele carriers might be ascribed to the higher serum E2 levels. PMID- 12767516 TI - Mutations of the BAK gene are infrequent in advanced gastric adenocarcinomas in Koreans. AB - The bcl-2 homologue antagonist/killer (BAK) is a potently apoptosis-inducing gene and plays an important role in modulating apoptosis in epithelial cells. We have analyzed the mutation of the entire coding region of BAK gene in 107 Korean advanced gastric adenocarcinomas by polymerase chain reaction-single strand conformation polymorphism and sequencing. Homozygous deletions were not found in these samples. Only three cases of 107 gastric adenocarcinomas (2.8%) exhibited the BAK mutations. Two of them exhibited missense mutations and the remaining one had a silent mutation. All of these mutations were exclusively detected in exon 2. Mutations in the BAK gene were observed only in advanced gastric adenocarcinomas with extensive metastases of regional lymph nodes. The data presented here suggest that the mutations of BAK gene rarely occurred in advanced gastric adenocarcinomas. PMID- 12767517 TI - Serum MUC5AC mucin as a potential marker for cholangiocarcinoma. AB - Aberrant expression of MUC5AC mucin is obvious in cholangiocarcinoma tissues, however, this mucin has never been detected in the serum. Using immunoblotting marked with antibody vs. MUC5AC core protein, we could detect MUC5AC mucin in the serum of 112 from 179 cholangiocarcinoma patients (62.6% sensitivity), two of the 62 with benign hepatobiliary diseases, six of the 60 with hepato-gastrointestinal cancer, and none in either the 60 active opisthorchiasis or 74 healthy persons. Detection of serum mucin in the serum of cholangiocarcinoma patients corresponded well to the MUC5AC expressed in individual tissues. Serum MUC5AC may be used to enhance the diagnostic accuracy of cholangiocarcinoma. PMID- 12767518 TI - Cytokine-suppressive anti-inflammatory drugs (CSAIDs) inhibit invasion and MMP-1 production of ovarian carcinoma cells. AB - The development of therapeutic strategies for inhibition of peritoneal dissemination and invasion would be central for the treatment of ovarian carcinoma. In the microenvironment of ovarian carcinomas, various inflammatory cytokines like tumor necrosis factor alpha (TNF-alpha) are present. In this study we investigated the role of inflammatory cytokines in the regulation of invasion of SKOV-3 ovarian carcinoma cells in-vitro. Treatment of cells with TNF-alpha or interleukin 1beta (IL-1beta) lead to increased phosphorylation of the stress activated p38 mitogen-activated protein kinase (p38MAPK). Furthermore, TNF-alpha as well as IL-1beta stimulated matrigel invasion of tumor cells. An inhibitor of stress-activated protein kinase pathways, the cytokine-suppressive anti inflammatory drug (CSAID) SB203580 inhibited invasion of cytokine-stimulated SKOV 3 cells. The MEK-1 inhibitor PD98059 similarly inhibited invasion of cytokine stimulated cells, but to a lesser extent. Expression of mRNA and protein levels of matrix metalloproteinase-1 (MMP-1) by SKOV-3 cells could be stimulated by inflammatory cytokines and inhibited by SB203580, and partially also by PD98059. Our results show that CSAIDs reduce invasion and MMP expression of ovarian carcinoma cells. Further studies are required to investigate whether inhibition of cytokine-induced signal transduction may be of value in therapy of ovarian carcinomas in-vivo. PMID- 12767519 TI - Malignant transformation of thyroid follicular cells by galectin-3. AB - Galectin-3, a beta-galactoside binding lectin, is highly expressed in thyroid carcinomas of follicular cell origin, whereas neither benign thyroid adenomas nor normal thyroid tissues express galectin-3. We previously showed that antisense inhibition of galectin-3 expression markedly reduced the malignant phenotype of thyroid papillary carcinoma cells. In the present study we transfected galectin-3 cDNA into TAD-2 normal thyroid follicular cells. Stable transfectants expressing galectin-3 acquired the phenotype of serum-independent growth, clonogenicity in soft agar, as well as loss of contact inhibition. We also compared the gene expression profile of the galectin-3 transfectants to that of the vehicle control, which revealed that a series of genes were differentially expressed between the two. They include proliferating cell nuclear antigen, replication factor C, and retinoblastoma genes that participate in G1-S transition. These results indicate the transformation of thyroid follicular cells by galectin-3 and possible involvement of galectin-3 in cell cycle. PMID- 12767520 TI - The insulin-like growth factor system and cancer. AB - The insulin-like growth factor (IGF) family of ligands, binding proteins and receptors is an important growth factor system involved in both the development of the organism and the maintenance of normal function of many cells of the body. The system also has powerful anti-apoptotic effects. More recently, evidence has accrued to demonstrate that the IGFs play an important role in cancer. Individuals with serum IGF-II levels in the upper quartile of the normal range (and IGF binding protein-3 levels in the lower quartiles) have a relative risk for developing breast, prostate, colon and lung cancer. IGF-II is commonly expressed by tumor cells and may act as an autocrine growth factor; occasionally even reaching target tissues and causing tumor-induced hypoglycemia. The IGF-I receptor is commonly (though not always) overexpressed in many cancers, and many recent studies have identified new signaling pathways emanating from the IGF-I receptor that affect cancer cell proliferation, adhesion, migration and cell death; functions that are critical for cancer cell survival and metastases. In this review, many aspects of the IGF system and its relationship to cancer will be discussed. PMID- 12767521 TI - Effect of dietary genistein on cell replication indices in C57BL6 mice. AB - The phytoestrogen and isoflavone, genistein, inhibited the activity of the DNA synthesis-related enzyme, topoisomerase-II (topo-II), altered cell-cycle traverse and produced cell death in cell culture models. In order to examine the potential effects of genistein on cell replication and cell death in an animal model, 8 week-old C57BL6 mice were fed either a control diet or one containing one of five doses (100-2000 ppm) of genistein for 28 days. At the end of the feeding period, both male and female mice were sacrificed and the serum isoflavone and aglycone levels determined by liquid chromatography with electrospray tandem mass spectrometry (LC-ES/MS/MS). Immunohistochemistry was utilized to measure the cell replication and cell death rates in the small intestine. Total isoflavone concentration increased from below the limits of detection (0.001 microM) in control animals to 0.28 microM in male and 0.54 microM in female mice fed the 2000 ppm diet. A decrease in the percentage of cells in G(0) and an increase in the percentage of cells in S-phase, consistent with topo-II-induced S-phase arrest, was found in the duodenum and jejunum of the small intestine. Thus, genistein appears to accumulate to a sufficient level to affect topo-II activity in the small intestine. PMID- 12767522 TI - No involvement of beta-catenin gene mutation in gastric carcinomas induced by N methyl-N-nitrosourea in male F344 rats. AB - The agent N-methyl-N-nitrosourea (MNU) is a direct acting carcinogen and induces well-differentiated adenocarcinoma on the rat gastric mucosa. In this study, 27 histopathologically verified gastric carcinomas induced in male F344 rats were analyzed for mutations in the N-terminal phosphorylation sites (codons 1-51) of the beta-catenin gene by using polymerase chain reaction (PCR)-single strand conformation polymorphism (SSCP) assays. In parallel studies, the specific localization of the beta-catenin protein was also examined by immunohistochemical analysis. No mutations in the beta-catenin gene were found in any of 27 gastric carcinomas induced by MNU. Immunohistochemical analysis resulted in the beta catenin protein to be localized in the plasma membrane but cytoplasmic and/or nuclear accumulation of beta-catenin was not identified in any of these carcinomas. These results suggest that mutations in the beta-catenin gene are less contributory to the development of rat gastric carcinomas induced by MNU. This animal model may provide a system for evaluating the mechanism of human gastric carcinogenesis that is not associated with beta-catenin gene mutations. PMID- 12767523 TI - In vitro sensitivity of hematopoietic progenitors to tiazofurin in refractory acute myeloid leukemia and in the blast crisis of chronic myeloid leukemia. AB - The effect of Tiazofurin (TR) on the in vitro growth of bone marrow (BM) and peripheral blood (PB) leukemic progenitors was investigated in 29 patients. Nineteen of the patients were suffering the blast crisis of chronic myeloid leukemia (bcCML) and ten patients refractory acute myeloid leukemia (AML). PB and BM mononuclear cells were cultured in methylcellulose alone or with concentrations of TR ranging between 10 and 200 microM. TR produced a dose dependent inhibition of colony forming unit (CFU)-blast growth in all the samples tested from BM and PB. The most effective concentrations of TR used were 150 and 200 microM, while concentrations of less than 50 microM TR were not adequate for 50% inhibition of cell growth (IC50). Differences were found in the response of CFU-blasts to TR related to the type of underlying leukemia. Inhibition of CFU blast growth was more pronounced in bcCML than in AML in both the BM and PB samples. The concentration of TR required to induce IC50 in bcCML was 50 microM, while the same effect in AML required a concentration of 150 microM. Analysis of the control samples also revealed that CFU-blasts from bcCML produced smaller numbers of colonies, though these differences were not statistically significant. It has therefore been demonstrated that TR has strong in vitro anti-leukemic activity, more pronounced in bcCML than in refractory AML. We thus feel this study gives further rationale for the clinical application of TR, and would strongly support this. PMID- 12767524 TI - Effects of FK228, a novel histone deacetylase inhibitor, on tumor growth and expression of p21 and c-myc genes in vivo. AB - In this study, we examined the effects of FK228 (FR901228, depsipeptide) on tumor growth and expression of p21 and c-myc genes in vivo. FK228 induced the expression of p21 mRNA and decreased c-myc mRNA in tumor xenograft sensitive to FK228. However, FK228 did not sufficiently modulate the expression of p21 mRNA and increased the expression of c-myc in tumor xenograft less sensitive to FK228. The modulation of p21 and/or c-myc genes may be critical for the marked antitumor activity of FK228 in vivo. PMID- 12767525 TI - Prognostic potential of glutathione S-transferase M1 and T1 null genotypes for gastric cancer progression. AB - To improve understanding of glutathione S-transferase (GST) behavior in terms of a development and prognostic factor for gastric adenocarcinoma, we investigated the association between the GSTM1 and GSTT1 null genotypes and gastric cancer risk or the prognostic value of the GSTM1 and GSTT1 null genotypes was evaluated. Using a polymerase chain reaction-based method, the frequencies of GSTM1 and GSTT1 genotypes and prognostic factors, such as staging, differentiation, and histologic type (intestinal vs. diffuse), were evaluated in 80 patients with curatively resected primary gastric adenocarcinoma. The frequencies of GSTM1 and GSTT1 null individuals were higher in the gastric cancer group, but the differences were not statistically significant (for GSTM1 null odds ratio (OR)=0.86; 95% confidence interval (CI)=0.49-1.51 and for GSTT1, OR=0.97; 95% CI=0.55-1.71). Since the GSTM1 and GSTT1 null genotypes are potential indicators of gastric adenocarcinoma, we examined the relationship between the GSTM1 and GSTT1 genotypes and prognostic factors. In terms of the histologically diffuse type of cancer, GSTM1 indicated an approximately 3.24-fold increase (OR=3.24; 95% CI=1.05-10.17). With respect to gastric cancer differentiation, the frequency of the GSTM1 null genotype was linked with a statistically significant increase in risk (3.42-fold) for the high-grade type (OR=3.42; CI=1.02-13.24). Our results indicate that there is no obvious relationship between GSTM1 and GSTT1 polymorphisms and the development of gastric cancer. However, in Korean gastric adenocarcinoma patients the GSTM1 null genotype appears to be associated with a poorer prognosis. PMID- 12767526 TI - Association of the genetic polymorphism in cytochrome P450 (CYP) 1A1 with risk of familial prostate cancer in a Japanese population: a case-control study. AB - Association between genetic polymorphisms of CYP1A1 and familial prostate cancer risk was examined by a case-control study of 185 individuals. Although the individual analysis of m1 or m2 genotype of CYP1A1 showed no significant association with prostate cancer risk, the presence of any mutated alleles significantly increased prostate cancer risk in comparison with wild-type genotypes by combination analysis (odds ratio [OR]=2.38; 95% confidence interval [CI]=1.72-3.29; P=0.0069). Furthermore, metastatic cancer had a significant association with mutated alleles of m1 and m2. These finding suggested that CYP1A1 polymorphisms has an association with prostate cancer risk, especially with progression of prostate cancer. PMID- 12767527 TI - A well-defined in vitro three-dimensional culture of human endometrium and its applicability to endometrial cancer invasion. AB - A three-dimensional (3-D) endometrium culture was established, in which human endometrial stromal cells embedded in a mixture of collagen I, a major component of extracellular matrix, and matrigel, a basement membrane material, supports the epithelial cells seeded on top of the collagen/matrigel matrix. The biological growth and differentiation of the epithelial cells were studied microscopically and immunohistochemically. Transmission electron microscopy showed a polarized columnar epithelium in monolayer with basally positioned nuclei. Scanning electron microscopy revealed a confluent epithelium with an abundance of microvilli and cilia as well as pinopodes on the apical surface. An immunohistochemical staining showed that integrin alpha1, alpha4, and beta3 were co-localized with cytokeratin, confirming the epithelial origin of the cells. In contrast, immunoreactivity against cyclooxygenase-1 or -2 was positive in both epithelial and stromal cells. When epithelial cells were replaced by KLE cells, an endometrial cancer cell of epithelial origin, invasion of KLE cells into the stromal fraction was observed. The invasion was closely correlated to expression of matrix metalloproteinases and their tissue inhibitors of metalloproteinases in a manner consistent with paracrine fashion. The present 3-D culture imitates the normal endometrium physiologically as well as morphologically, thus provides an excellent in vitro tissue suitable for reproducing in vivo physiological processes, including endometrial cancer invasion. PMID- 12767528 TI - Enhanced MTT-reducing activity under growth inhibition by resveratrol in CEM-C7H2 lymphocytic leukemia cells. AB - Inhibition of proliferation by resveratrol of CEM-C7H2 lymphocytic leukemia cells was paradoxically associated with an enhanced cellular 3-[4,5-dimethylthiazol-2 yl]-2,5-diphenyltetrazolium bromide (MTT)-reducing activity. This phenomenon was most pronounced at the sub-apoptotic concentration range of 5-20 microM resveratrol. The results of our study show that the MTT-reducing activity can be increased by the polyphenolic antioxidant resveratrol without a corresponding increase in the number of living cells and that this occurs at a concentration range of the antioxidant which is not sufficient to induce apoptosis but suffices to slow down cell growth. This phenomenon appears to be restricted to proliferation inhibitors with antioxidant properties and is cell type-specific. Thus, in determining the effects of flavonoids and polyphenols on proliferation, in certain cell types this might represent a pitfall in the MTT proliferation assay. PMID- 12767529 TI - LIGHT sensitizes IFNgamma-mediated apoptosis of MDA-MB-231 breast cancer cells leading to down-regulation of anti-apoptosis Bcl-2 family members. AB - LIGHT is a new member of the tumor necrosis factor superfamily, which binds to lymphotoxin beta receptor, herpes virus entry mediator, or TR6. This work was carried out to elucidate the molecular mechanism of LIGHT-sensitized, interferon gamma (IFNgamma)-mediated apoptosis of MDA-MB-231 cells. It was revealed that LIGHT treatment resulted in down-regulation of anti-apoptosis Bcl-2 family member: Bcl-2, Bcl-X(L), Bag-1, and Mcl-1; up-regulation of pro-apoptosis Bcl-2 family member: Bak and Ser (112)-phosphor-Bad; down-regulation of pro-apoptosis Bcl-2 member Bax; the other pro-apoptosis member Bid remains unaltered. LIGHT treatment also resulted in activation of caspase-3, caspase-6, caspase-7, caspase 8, caspase-9, DFF45, and PARP. However, caspase activation and caspase activity, especially caspase-3 activity, is not required for LIGHT-induced apoptosis of MDA MB-231 cells, since caspase-3 inhibitor, benzyloxycarbonyl-Asp-Glu-Val-Asp fluoromethylketone, and a broad range caspase inhibitor, benzyloxycarbonyl-val ala-asp-fluoromethylketone failed to block the apoptosis induced by LIGHT and IFNgamma in MDA-MB-231 cells. In summary, LIGHT-sensitized IFNgamma-mediated apoptosis of MDA-MB-231 cells is probably through down-regulation of anti apoptosis Bcl-2 family members; it could be caspase (especially caspase-3) independent, even though extensive caspase activation was observed. PMID- 12767531 TI - Gene expression profile of human cells irradiated in G1 and G2 phases of cell cycle. AB - The response of cells to ionizing radiation has long been known to result in alterations in gene expression. Here we examined the expression patterns of synchronized HeLa cells in G1 or G2, after exposure to 2Gy of X-rays on microarrays allowing evaluation of over 12,000 human genes. The analysis showed induction of at least twofold for 257 different genes, after irradiation of cells in G1 and down-regulation of 42. The altered transcription patterns involved genes belonging to cell cycle pathways, DNA repair, oncogenes, histones, heat shock genes, mitochondrial and ribosomal proteins, transcription and translational regulators and genes encoding cytoskeleton components. HeLa cells are deficient in p53 and none of the induced genes have previously been identified as regulated by p53. One hundred and seventy-seven genes were up regulated and 26 genes were down-regulated after irradiation in G2. Surprisingly most of the up-regulated genes in both phases of the cell cycle encode ribosomal proteins. These studies have revealed cell cycle dependence on radiation-induced gene induction and have permitted the identification of previously known as well as unknown genes associated with radiation-induced altered expression. PMID- 12767530 TI - Nuclear delivery of p53 C-terminal peptides into cancer cells using scFv fragments of a monoclonal antibody that penetrates living cells. AB - scFv fragments of a monoclonal antibody that penetrates living cells and localizes in nuclei were designed as fusion proteins with C-terminal p53 peptides and tested for restoring p53 function in p53 mutant cancer cells. scFv fragments transported a 30-mer C-terminal peptide of p53 into cancer cells and induced cellular cytotoxicity in contrast to scFv fragments alone and other scFv-p53 fusion peptides. Cellular toxicity was not observed with scFv fragments containing a single mutation in VH that prevented antibody penetration. Our results demonstrate the potential efficacy of antibody scFv fragments as a nuclear delivery system in cancer cells. PMID- 12767532 TI - Orthotopic implantation of a colon cancer xenograft induces high expression of cyclooxygenase-2. AB - The aim of the present study is to investigate the relation of cyclooxygenase (COX)-2 with liver metastasis. Pieces of human colon cancer xenograft were implanted orthotopically (CI), as well as intraperitoneally, and subcutaneously. Liver metastasis developed most frequently, and the COX-2 expression of both mRNA and protein of the tumors was the most dominant in the CI group. In contrast, the expression of COX-1 or vascular endothelial cell growth factor mRNAs had no significant differences among the groups. The intensity of COX-2 mRNA was negatively correlated with the apoptotic index. In conclusion, COX-2 plays an important role in organ-specific metastasis of the colon cancer. PMID- 12767534 TI - Current concepts of cardiovascular diseases in diabetes mellitus. AB - The incidence of diabetes has reached epidemic proportions across the world. In patients with diabetes, there is a two to four times increased risk of developing coronary artery disease (CAD). Diabetes seems to eliminate the protective benefits of hormones in women against CAD. Patients with type II diabetes also have hypertension, dyslipidemia, obesity, endothelial dysfunction and prothrombotic factors, called 'the metabolic syndrome'. Not only the incidence of CAD is higher in diabetes, the mortality of the diabetic patients after a cardiac event is significantly increased as compared to non-diabetics, including sudden death. Although in the past 35 years there has been a decline in the rate of death due to CAD in the general population, this has not been seen among patients with diabetes. Primary prevention can play an important role in decreasing the incidence of CAD in diabetic patients. Aggressive treatment of hyperlipidemia and hypertension is essential. Recent knowledge about the protective effects of aspirin, statins, angiotension converting enzyme inhibitors, and glitazones in the diabetic patients, if used appropriately will go a long way in primary and secondary prevention of CAD in patients with diabetes. PMID- 12767535 TI - Percutaneous coronary revascularization improves the prognosis of patients with cardiogenic shock in acute coronary syndrome: a chronological study. AB - BACKGROUND: Cardiogenic shock complicating acute coronary syndrome (ACS) implies grim prognosis with conventional management. Previous studies of coronary intervention yielded controversial results and were rarely analyzed chronologically. This study was to determine the impact of percutaneous coronary revascularization on outcome by studying two time periods 5 years apart in which the revascularization was more frequent and techniques more refined in the later period. METHODS AND MATERIALS: All patients admitted to the intensive or coronary care unit for ACS in two 1.5-year study periods (Period I: Jan 1994-Jun 1995, Period II: Oct 1999-Apr 2000) were retrospectively screened. Patients who met strict criteria of cardiogenic shock within 24 h of ACS were enrolled. The demographics, management and in-hospital/3-month outcomes were analyzed. RESULTS: Thirty-seven patients (33M/4F, aged 65+/-8 years) were enrolled in Period I and 32 patients (25M/7F, aged 68+/-13 years) in Period II. The incidence of cardiogenic shock was 11.8 and 9.3%, respectively. The demographics were similar except patients in Period II were older. Significantly more coronary angiography and interventions were done in the later period. The in-hospital (68 vs. 44%, P=0.047) and 3-month mortalities (70 vs. 44%, P=0.03) were significantly reduced in Period II. The in-hospital survivors in two study periods differed only in use of coronary angiography (94 vs. 50%, P=0.005) and interventions (83 vs. 33%, P=0.005) but not others. CONCLUSIONS: Percutaneous coronary revascularization does improve the clinical outcome of cardiogenic shock when analyzed chronologically. This treatment is warranted in every such patient in the interventional era. PMID- 12767533 TI - Effect of H(2)O(2) on cell cycle and survival in DNA mismatch repair-deficient and -proficient cell lines. AB - Patients who develop tumors with Lynch syndrome, which is caused by mutational inactivation of the DNA mismatch repair (MMR) system, have a relatively favorable prognosis compared to patients who develop sporadic tumors. Paradoxically, DNA MMR-deficient cells are resistant to many chemotherapeutic agents, and are capable of bypassing the G2/M checkpoint in vitro. Colon cancers that develop in the setting of Lynch syndrome show an abundant recruitment of immune cells into tumor tissues, which might be expected to increase oxyradical formation, and make the tumor cells more vulnerable to cell death. We examined the chemosensitivity and cell cycle response to oxidative stress in several MMR-deficient (HCT116, SW48, and DLD1) and -proficient (CaCo2, SW480, and HT29) colorectal cancer cell lines. H(2)O(2) induced a G2/M cell cycle arrest in both MMR deficient and proficient cell lines, however MMR-deficient cell lines were more sensitive to H(2)O(2) toxicity, and the response was more prolonged in MMR-deficient cells. Interestingly, human MutL-homologue (hMLH1-)defective HCT116 and hMLH1-restored HCT116+ch3 cell lines responded to H(2)O(2) with the same degree of G2/M arrest. The survival response of HCT116+ch3 was nearly identical to that of hMLH1 defective HCT116+ch2, although better than the response observed in HCT116 cells. In conclusion, greater cellular sensitivity and G2/M arrest in response to oxidative stress in MMR-deficient colorectal cancer cells could be one of the reasons for the more favorable prognosis seen in patients with Lynch syndrome. However, this sensitivity appears not to be a direct result of a deficient MMR function, but is more likely attributable to spectrum of target gene mutations that occurs in MMR-deficient tumors. PMID- 12767536 TI - Prognosis of patients with unstable angina and low-risk exercise test: significance of ST-segment depression on the admission ECG. AB - The presence of ST-segment depression on the admission electrocardiogram (ECG) is an important predictor of poor outcome in patients with unstable angina. On the other hand, patients with unstable angina who undergo a low-risk exercise test are supposed to have a favorable prognosis. The objective of the study was to determine the prognostic significance of ST-segment depression on the admission ECG in patients with unstable angina who undergo an exercise test that indicates a low risk of events. An interpretable exercise test was performed in 257 patients with primary unstable angina. A low-risk exercise test was completed by 156 (60%) patients and medical therapy was planned for all these patients. A multivariate analysis was performed in order to determine the independent predictors of events (cardiac death, nonfatal acute myocardial infarction, or admission for unstable angina) during a 12-month follow-up. Among patients with a low-risk exercise test, there were no significant differences between patients with and without ST-segment depression on the presenting ECG with regard to event rate (34 vs. 29%, P=NS). In multivariate analysis, ST-segment depression was not related to a higher incidence of events. Our findings appear to indicate that the presence of ST-segment depression on the admission ECG loses its prognostic significance in patients with primary unstable angina if they complete a low-risk exercise test. PMID- 12767537 TI - Antiarrhythmic effect of magnesium sulfate after open heart surgery: effect of blood levels. AB - BACKGROUND: Arrhythmias following cardiac surgery is still a difficult complication to treat. Magnesium sulfate is an effective antiarrhythmic agent with negligible side effects. In this study, effects of magnesium sulfate as a first line antiarrhythmic agent was compared with results of two different well accepted antiarrhythmic agents. METHODS: One hundred patients with arrhythmia were prospectively randomized to a study and a control group. Lidocaine and amiodarone were accepted as standard antiarrhythmic agents. Patients in study group were received magnesium sulfate routinely as a first line antiarrhythmic agent. Unresponsive arrhythmias were treated with standard antiarrhythmic agents. Control group patients received only standard antiarrhythmics. RESULTS: Magnesium sulfate alone was effective in 56% of the study group whereas 74% of the control group were responsive to standard antiarrhythmics (P=n.s.). In study group, a subgroup analysis according to blood levels of Mg2+ revealed that magnesium sulfate was more effective in patients with low Mg2+ levels (63% for low Mg2+ levels, 55% for normal Mg2+ levels, 36% for high Mg2+ levels) and ventricular arrhythmias (60% for ventricular and 40% for supraventricular arrhythmias), without statistical significance. CONCLUSIONS: Magnesium sulfate is an effective and safe antiarrhythmic agent for arrhythmias developed after open-heart surgery. Its antiarrhythmic effect may relate to its pharmacological properties and unrelated to normalization of the circulating magnesium concentrations. We recommend its use as a first line antiarrhythmic agent without routine measurement of blood levels. PMID- 12767538 TI - Restoration of nocturnal dip in blood pressure is associated with improvement in left ventricular ejection fraction. A 1-year clinical study comparing the effects of amlodipine and nifedipine retard on ambulatory blood pressure and left ventricular systolic function in Chinese hypertensive type 2 diabetic patients. AB - We assessed the effects of amlodipine and nifedipine retard on 24-h blood pressure (BP) control, nocturnal fall in BP and their significance on left ventricular systolic functions in 54 Chinese hypertensive type 2 diabetic patients. Patients being recruited were openly randomised to amlodipine or nifedipine retard. Ambulatory 24-h BP and echocardiogram (in 42 patients) were measured before and 1 year after treatment. At the end of study, there was 17% reduction in systolic BP; 17% reduction in diastolic BP and 12% reduction in mean arterial pressure (MAP) (no difference between amlodipine and nifedipine). Of the 42 subjects underwent echocardiograms, eight became 'new-dippers' at the end of study (non-dippers before treatment and restored nocturnal fall of MAP> or =10% after treatment). The other 34 patients were either non-dippers before and after treatment (n=27); dippers before and after treatment (n=3) or dippers before treatment and non-dippers after treatment (n=4). The eight 'new-dippers' had improved ejection fraction (69.6+/-7.2 to 75.8+/-7.4%, P<0.05) and increased left ventricular diastolic diameter (43.7+/-7.9 to 47.9+/-8.8 mm, P<0.05) after the 1 year treatment of calcium antagonist. Compared to the other 34 subjects, the eight 'new-dippers' showed significant improvement in ejection fraction (9.4+/ 10.9 vs. -1.2+/-11.8%, P<0.05). In conclusion, both amlodipine and nifedipine retard are effective in controlling the 24-h BP in Chinese hypertensive type 2 diabetic patients. For those who have restored nocturnal dip in BP have significantly increased left ventricular systolic ejection fraction after 1-year treatment of long acting calcium antagonists. The clinical significance and underlying mechanisms require further studies. PMID- 12767539 TI - Multislice spiral computed tomography for the detection of coronary stent restenosis and patency. AB - BACKGROUND: Multislice spiral computed tomography (MSCT) has evolved as a new promising method for non-invasive visualization of the coronary arteries and detection of native coronary artery stenosis. We determined the value of MSCT to non-invasively detect significant in-stent restenosis after coronary artery stenting. METHODS: Twenty patients (age 56.3+/-8.6 years) were investigated by MSCT (4x1 mm cross-sections, 500 ms tube rotation, table feed 1.5 mm/rotation, intravenous contrast agent, retrospectively ECG-gated image reconstruction) at a mean interval of 9.6+/-4.2 months after coronary stent implantation. Results were compared with conventional quantitative coronary angiography (QCA). A total number of 32 stents were studied, four different stent types were evaluated. RESULTS: QCA showed in-stent restenosis >50% diameter stenosis in five (16%) stents. Using MSCT it was impossible in all stents, irrespective of stent type or diameter, to directly visualize the stent lumen due to partial volume effects and beam hardening. MSCT allowed the visualization of the coronary vessel proximal and distal to the stent. This allowed confirmation of stent patency in 18/18 cases and correct identification of total stent occlusion in two patients. CONCLUSIONS: MSCT allows no direct visualization of coronary in-stent restenosis, but it correctly differentiates between stent patency and stent occlusion. The reasons are mainly partial volume effects and beam hardening, which are induced by the stent material. PMID- 12767540 TI - Prospective evaluation of hydroperoxide plasma levels and stable nitric oxide end products in patients subjected to angioplasty for coronary artery disease. AB - BACKGROUND: Oxidative stress appears to be involved in several processes that contribute to atherogenesis and restenosis following vascular intervention. METHODS: The aim of our study was to evaluate prospectively the plasma concentrations of a hydroperoxide (ROOH) and nitric oxide end product (NO(x)) in patients subjected to coronary angioplasty (PTCA) and routine control angiography 6 months after the initial procedure. We prospectively studied 48 consecutive patients (39 men, nine women, mean age 52 years) with stable angina who underwent successful elective angioplasty. A vascular segment was considered successfully treated when the residual luminal narrowing in the dilated segment immediately after angioplasty was <50%. Angiographic follow-up was obtained in all of the patients. Plasma samples were drawn at baseline (before angioplasty) and serially after angioplasty (1, 3 and 6 months afterwards). Hydroperoxides were determined by the FOX II assay (ferrous oxidation in xylenol orange, Pierce Rockford, IL). Nitrate was converted in the presence of NO3 reductase. The Griess reagent was used for the measurement of NO2. RESULTS: The overall angiographic restenosis rate was 35%. There were no significant differences in clinical variables between the patients with or without restenosis. The baseline levels (0.8+/-0.09 vs. 0.6+/-0.2 micromol/l) as well as the concentrations of authentic lipid hydroperoxide in plasma after 1 month (0.7+/-0.09 vs. 1.0+/-0.2 micromol/l) and 6 months (0.8+/-0.1 vs. 1.0+/-0.2 micromol/l) were similar in both groups. Three months after the angioplasty a significant increase in the ROOH level was noticed in the patients with restenosis (0.9+/-0.1 vs. 1.4+/-0.2, P=0.04). Plasma levels of NO(x) were similar in both groups at baseline (23.6+/-2.1 vs. 22.7+/-2.6 micromol/l) and 1 month after procedure (24.4+/-2.2 vs. 23.4+/-3.3 micromol/l). However, in patients with restenosis significant decreases in stable NO end products were observed 3 and 6 months after PTCA (18.1+/-1.5 vs. 13.3+/-1.7, P=0.04; 14.2+/-1.0 vs. 8.7+/-1.3, P=0.02, respectively). CONCLUSIONS: In patients with angiographic restenosis a significant increase in lipid peroxidation accompanied by a reduction in the stable end products of nitric oxide in plasma is observed several months after PTCA. PMID- 12767541 TI - Does a prolonged QT peak identify left ventricular hypertrophy in hypertension? AB - BACKGROUND: Left ventricular hypertrophy is a powerful predictor of death. Hypertensive subjects with left ventricular hypertrophy can have increased QT (end) dispersion, which is associated with cardiac death. Despite its prognostic value, QT (end) dispersion is not widely used. QTp (i.e. start of QRS to peak of T wave) is easier to measure. Therefore, we tested the hypothesis that long QT peak was associated with left ventricular hypertrophy and assessed its cost effectiveness at diagnosing left ventricular hypertrophy. METHODS: ECGs and echocardiograms were recorded in 47 hypertensive patients. The onset of the QRS complex and peak of T wave of lead I of each subject's ECGs were digitised by one observer blind to results of the echocardiogram. Receiver-operator characteristics curves were plotted to determine the sensitivity and specificity of different cut-off values of QT peak at predicting left ventricular hypertrophy (defined as left ventricular mass index> or =134 g/m2 in male, > or =110 g/m2 in female). RESULTS: The heart-rate corrected QT peak of lead I correlated with left ventricular mass index (r=0.45, P=0.002). If all patients with a prolonged QT peak (> or =300 ms) had an echocardiogram, then no cases of left ventricular hypertrophy would be missed (100% sensitive). This novel ECG criterion not only had better positive and negative predictive values than the Sokolow-Lyon voltage criteria, but also resulted in more cost-effective resource use (< pound 370 vs. pound 1750/case of left ventricular hypertrophy detected). CONCLUSION: If the results of this small pilot study are confirmed in larger studies, then measuring QT peak of lead I may become a cost-effective way of identifying hypertensives who are likely to have echocardiographic left ventricular hypertrophy. PMID- 12767542 TI - Homogenous distribution of fast response action potentials in canine pulmonary vein sleeves: a contradictory report. AB - Pulmonary veins may serve as source of ectopic focus (or foci) in initiating atrial tachyarrhythmias in human beings. However, the animal model for such focal atrial fibrillation is still lacking and cellular mechanism for arrhythmias remains to be studied. Recently, a series of reports of cellular electrophysiological characterization of pulmonary vein sleeves demonstrated an extremely high incidence of automaticity (varied from 40 to 76%) and triggered activity (from 0 to 44%) in normal healthy control dogs and rabbits. The present study was therefore designed to re-investigate the cellular electrophysiological properties of canine pulmonary veins. Intracellular action potentials were characterized in pulmonary vein sleeves in 50 normal healthy dogs. Conventional glass microelectrode recording technique was used. Experiments were focused on the incidence of automaticity and triggered activity in pulmonary vein sleeve tissues. Surprisingly, our results showed that all pulmonary vein sleeves tissues in these dogs displayed fast-response action potentials under the well-controlled experimental condition (100%, n=50). No spontaneous pacemaking activities, early or delayed afterdepolarisations were observed (0%, n=50). No high-frequency spikes or irregular rhythm could be recorded in all experiments (0%, n=50). Both the frequency response and membrane responsiveness of the pulmonary vein action potentials were characterized. No electrophysiological inhomogeneity between the distal and the proximal part of the sleeves was observed. In conclusion, canine pulmonary vein sleeves do not display arrhythmogenic activities under normal physiological conditions. The possible explanations for the controversy in pulmonary veins electrophysiology were discussed. PMID- 12767543 TI - N-terminal proatrial natriuretic peptide in primary care: relation to echocardiographic indices of cardiac function in mild to moderate cardiac disease. AB - AIMS: In patients with chronic heart failure plasma levels of N-terminal proatrial natriuretic peptide (Nt-proANP) correlate to cardiac filling pressures. The aim of the present study was to examine the relation between Nt-proANP plasma levels and echocardiographic indices of cardiac dysfunction in patients recruited from primary care. METHODS AND RESULTS: After clinical examination by the primary care physician, the patients were referred to one of two centres for echocardiography and blood sampling. In patients with mild to moderate symptoms of heart failure (n=52) and in asymptomatic patients with long-standing hypertension (n=46) or previous myocardial infarction (n=97), peptide levels were most closely related to parameters of left atrial wall stress. Patients who according to echocardiographic predefined criteria had diastolic or systolic dysfunction had two- and three-fold higher Nt-proANP than controls. According to receiver operating curve (ROC) analysis, Nt-proANP measurements were helpful in ruling out left ventricular systolic dysfunction, but not diastolic dysfunction. CONCLUSION: In patients with mild to moderate cardiac disease, Nt-proANP plasma concentration was related to increased atrial wall stress. Peptide measurement could assist in ruling out the presence of LV systolic dysfunction, but was otherwise of limited value when used for diagnostic subgrouping into echocardiographically determined function categories. PMID- 12767544 TI - Impact of early changes in left ventricular filling pattern on long-term outcome after acute myocardial infarction. AB - BACKGROUND: In patients with heart failure due to chronic ischemic heart disease improvement of diastolic function indicates improved survival and a reduced morbidity, but whether this is also the case after acute myocardial infarction is not known. METHODS: To assess the prognostic importance of changes in left ventricular filling pattern, assessed with mitral deceleration time and colour M mode flow propagation velocity, on cardiac death and readmission due to heart failure serial Doppler echocardiography was carried out in 103 patients with a first myocardial infarction. Based on echocardiography on hospital admission and after 1 month, patients were divided into three groups: group A (n=29) comprised patients with normal filling at either examination, group B (n=29) comprised patients with improvement of initially abnormal filling, and group C (n=45) patients with deterioration or no change of an abnormal filling pattern. RESULTS: One-year survival free of cardiac death or hospitalisation for heart failure was 97% in group A, 86% in group B and 64% in group C (P<0.0001). In Cox analysis persistence of abnormal filling or deterioration of left ventricular filling was still a predictor of the combined endpoint (risk ratio 4.4, 95% CI 1.8-12.0, P=0.003) after adjustment of LV filling on admission, left ventricular systolic function and clinical variables. Serial analyses of left ventricular systolic function demonstrated a significant improvement after 1 year in ejection fraction in groups A and B, whereas ejection fraction remained unchanged in group C. CONCLUSION: Patients with a persistently abnormal or a deterioration of left ventricular filling pattern as opposed to improved or normal filling are at increased risk of cardiac death and readmission due to heart failure after acute myocardial infarction. PMID- 12767545 TI - Long-term outcome of acute myocarditis is independent of cardiac enzyme release. AB - BACKGROUND: There are few data concerning prognostic markers of acute myocarditis. The purpose of this study was to assess the prognostic value of initial measurements of creatine kinase (CK), cardiac troponin I (cTnI) and myoglobin as regards late recovery of the left ventricular ejection fraction on follow-up. METHODS: A total of 22 patients (53+/-15 years old, 11 female) with acute myocarditis were followed up in a prospective observational study. Of these, 11 (50%) showed a history of acute infection prior to hospitalisation and seven (32%) had pericardial effusion. The median ejection fraction during the acute phase was 47+/-17%; after a mean follow-up of 119+/-163 days it improved to 60+/-9% (P<0.001). Considering maximal CK-rise values of 641+/-961 U/l (P=0.38), cTnI-rise values of 3.7+/-8.6 microg/l (P=0.16) and myoglobin values of 7.4+/-12 nmol/l (P=0.69), there was no correlation between initial cardiac enzyme levels and the initial and late left ventricular ejection fraction. CONCLUSION: After acute myocarditis, there is late recovery of left ventricular ejection fraction, which is independent of the initial myocardial damage measured by cardiac enzyme release. PMID- 12767546 TI - Long-term results after partial left ventriculectomy. AB - BACKGROUND: Long-term clinical, echocardiographic and hemodynamic effects after partial left ventriculectomy (PLV) and predictors of outcome have been determined. METHODS: Between January 1995 and July 1999, PLV was performed in 39 patients. In 15 patients the etiology of heart failure was idiopathic dilated cardiomyopathy (DCMP), 19 patients had ischemic cardiomyopathy (ICMP) and five patients had valvular cardiomyopathy. Concomitant procedures included coronary artery bypass grafting in 16 patients, mitral valve repair in 33 patients and aortic valve replacement in five patients. All patients belonged in New York Heart Association (NYHA) functional class III or IV. Mean follow-up was 663+/-514 days. Clinical, echocardiographic and hemodynamic assessments and metabolic stress testing were performed preoperatively, within 30 days postoperatively and 6, 12 and 24 months after the operation. RESULTS: Actuarial survival was 64% after 1 year, 55% after 2 years and 44% 3 years after the operation. In patients with ICMP as well as in patients with DCMP actuarial 1 year survival was 60%. At 2-year follow-up NYHA functional class was improved significantly (P<0.05), but LV ejection fraction, LV end-diastolic diameter, cardiac index and peak oxygen consumption did not differ significantly from preoperative values. Analysis of factors influencing postoperative outcome indicated that decreased left ventricular wall thickness and a failure to increase the stroke volume index as a response to preoperative dobutamine administration were associated with postoperative mortality. CONCLUSIONS: PLV is associated with considerable postoperative mortality and lacking long-term improvement of cardiac performance. PMID- 12767547 TI - Electrocardiographic prediction of the development and site of acute myocardial infarction in patients with unstable angina. AB - BACKGROUND: ST-T changes on 12-lead electrocardiograms (ECGs) in patients with unstable angina (UA) have limited values for prediction of subsequent acute myocardial infarction (AMI). The aim of the present study is to obtain more useful ECG signs during UA in predicting the risk and the site of AMI. METHODS: ECGs were recorded from 238 consecutive patients with UA; 149 developed AMI, whereas 89 did not in the following 60 days after the UA episodes. P, ST-T and U wave changes in these AMI and non-AMI patients were analyzed retrospectively. Three groups of ECG leads were referred to reflect ischemic changes of anterior (V1-V5), lateral (I, aVL and V6) and inferior (II, III, and aVF) left ventricular walls. To explore the site-dependent predictors, the 149 AMI patients were divided into two groups; group A/L with anterior, antero-septal, apical or lateral AMI, versus group I/P with inferior or posterior AMI. RESULTS: ST depression > or =1 mm and abnormal T wave or U wave changes and P wave abnormalities were observed more frequently in AMI patients than non-AMI patients. On multivariate analysis, an independent ECG finding of the development of AMI was a biphasic U wave (odds ratio (OR) 5.4, 95% confidence interval (CI), 1.9-15.6, P=0.002) in the anterior leads. An inverted T wave (OR 5.1, 95%CI, 1.7 15.5, P=0.0036) and a biphasic U wave (OR 6.0, 95%CI, 2.2-16.1, P=0.0004) in the anterior leads were independent predictors of AMI in group A/L. There was no independent ECG predictor of group I/P. CONCLUSIONS: Biphasic U wave in anterior leads during UA is a useful ECG observation in the risk stratification of subsequent AMI. The independent ECG predictors of antero-lateral MI are inverted T wave and biphasic U wave. PMID- 12767548 TI - Amiodarone for pharmacological cardioversion of recent-onset atrial fibrillation. AB - The efficacy and safety of amiodarone for pharmacological cardioversion of recent onset atrial fibrillation was examined by reviewing the trials on the subject identified through a comprehensive literature search. Amiodarone has been used both intravenously (i.v.) and orally for the pharmacological cardioversion of recent-onset atrial fibrillation. Intravenous amiodarone has been used as a bolus only or as a bolus followed by a continuous i.v. infusion until conversion or up to 24 h. The dose of i.v. bolus given ranged from 3 to 7 mg/kg body weight and that of infusion from 900 to 3000 mg/day. The efficacy reported is 34-69% with the bolus only regimens, and 55-95% with the bolus followed by infusion regimens. Only the higher dose (>1500 mg/day) amiodarone is superior to placebo in converting recent-onset atrial fibrillation to sinus rhythm. The highest 24-h conversion rates have been reported with the i.v. regimen of 125 mg/h until conversion or a maximum of 3 g and the oral regimen of 25-30 mg/kg body weight administered as a single loading-dose (>90% and >85%, respectively). Most of the conversions occur after 6-8 h of the initiation of therapy. Predictors of successful conversion are shorter duration of atrial fibrillation, smaller left atrial size, and higher amiodarone dose. Amiodarone is not superior to the other antiarrhythmic drugs conventionally used for the pharmacological cardioversion of recent-onset atrial fibrillation but is relatively safe in patients with structural heart disease and in those with depressed left ventricle function. Therefore, amiodarone could be used particularly in patients with structural heart disease and in those with left ventricular systolic dysfunction as the use of class IC drugs, propafenone and flecainide, for cardioversion of atrial fibrillation is contraindicated in such patients. PMID- 12767549 TI - Subclinical skeletal muscle abnormalities in patients with hypertrophic cardiomyopathy and their relation to clinical characteristics. AB - BACKGROUND: Some mutations of cardiac sarcomeric proteins causing hypertrophic cardiomyopathy (beta-myosin heavy chain) are associated with skeletal muscle fiber dysfunction, while subclinical skeletal myopathy can be diagnosed by electromyography (EMG) in a substantial proportion of hypertrophic cardiomyopathy patients. METHODS: In 49 consecutive, unrelated patients with hypertrophic cardiomyopathy, conventional EMG of deltoid, vastus lateralis, tibialis anterior and soleus muscles was performed. No patient had clinically detectable muscle weakness. We compared the clinical and echocardiographic characteristics between patients with normal and patients with myopathic EMG. RESULTS: Myopathic EMG findings were demonstrated in 13 patients (26.5%), 26 patients (53.1%) had normal findings and 10 patients (20.4%) had indeterminate recordings. There was no significant difference in mean age, maximum wall thickness, left ventricular fraction shortening, NYHA class, the existence of left ventricular outflow tract obstruction, syncope, or the occurrence of nonsustained ventricular tachycardia in the Holter recording among the three groups. Comparison between the myopathic and the normal group revealed that nine patients from the latter (34.6%) had a positive history of sudden death in the family, whereas no patient had such a history in the former group (P=0.015). CONCLUSION: The higher prevalence of a family history of sudden death in patients with normal EMG, although not thoroughly explained by our data, may reflect differences in the genetic substrate produced by the higher prevalence of high-risk mutations that are not expressed in skeletal muscle (e.g. troponin T). Further evaluation in genotyped patients is warranted. PMID- 12767550 TI - Lipid profiles and endothelial function with low-dose hormone replacement therapy in postmenopausal women at risk for coronary artery disease: a randomized trial. AB - AIMS: To compare the effect of low (0.3 mg) and commonly prescribed (0.625 mg) doses of conjugated equine estrogens (CEE) on brachial artery flow-mediated dilation and lipid profiles. METHODS AND RESULTS: Twenty-five postmenopausal women (mean age, 65+/-6 years) at risk for coronary artery disease (CAD) (> or =2 established risk factors) entered a double-blind crossover study. Brachial artery endothelial function was evaluated by means of high-resolution vascular echography. Both CEE doses significantly decreased total cholesterol (-13%, 0.3 mg; -15%, 0.625 mg), low-density lipoprotein-cholesterol (LDL-C) (-15%, 0.3 mg; 16%, 0.625 mg), and lipoprotein(a) (-28%, 0.3 mg; -39%, 0.625 mg) values from baseline levels. Both treatments increased high-density lipoprotein-cholesterol (HDL-C) (5%, 0.3 mg; 7%, 0.625 mg) and triglycerides (3%, 0.3 mg; 8%, 0.625 mg). There was no dose effect for changes in the LDL-C/HDL-C ratio (-21%, 0.3 mg; 23%, 0.625 mg). Both doses improved brachial artery dilation during reactive hyperemia by 63% over baseline. CONCLUSION: In women at risk for CAD, low-dose hormone replacement treatment (HRT) improves lipid profiles and brachial artery endothelial function comparably to the most commonly prescribed dose. The benefit:risk ratio of low-dose HRT provides an attractive option for postmenopausal women at risk for CAD. PMID- 12767551 TI - Association of two angiotensinogen gene polymorphisms, M235T and G(-6)A, with chronic heart failure. AB - The aim of the study was to focus on the relationship between the angiotensinogen (AGT) gene polymorphisms, M235T and promoter G(-6)A, and chronic heart failure in the Czech population. A total of 158 patients with chronic heart failure (functional class NYHA II-IV, ejection fraction <40%, cardiothoracic index >50%) were compared with a control group of 200 subjects of similar age and sex distribution, without any personal history of cardiovascular diseases. The AGT gene polymorphisms were detected by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) methods. No significant differences in distributions of AGT genotypes between patients with chronic heart failure (CHF) and controls were found. The differences in distributions of alleles in AGT M235T (P(a)=0.02) and genotypes in AGT G(-6)A (P(g)=0.017) were found within women groups. Within CHF patients the distribution of AGT G(-6)A genotypes was not consistent with Hardy-Weinberg equilibrium (P=0.0001). We found significant relative risk of CHF in the GGMT genotype, OR=2.63 with 95% CI 1.39 4.95, P(corr)=0.01 (in the male group OR=1.83, 95% CI 0.92-3.66, P(corr)=0.3; in the female group OR=15.5, 95% CI 1.86-129.42, P(corr)=0.008). We provide evidence of increased risk in subjects with the GGMT variant of associated genotype of AGT gene for CHF, especially of fifteen-fold risk of this variant in women. PMID- 12767552 TI - Prevalence of Chlamydia pneumoniae in the atherosclerotic plaque of patients with unstable angina and its relation with serology. AB - BACKGROUND: Chlamydia pneumoniae has been associated with coronary artery disease by both seroepidemiological studies, and by direct detection of the micro organism in atherosclerotic lesions. This bacteria could play a potential role in the development of acute coronary events. We examined coronary arteries from patients with unstable angina in order to verify an endovascular presence of C. pneumoniae, and to determine if there is any relationship between serology of acute infection by this pathogen and its presence inside the atherosclerotic plaque of these patients. METHODS: We analysed a total of 76 atherosclerotic plaques obtained from 45 patients who underwent coronary artery bypass surgery. In all patients unstable angina was present within the prior 3 weeks. The presence of C. pneumoniae in the plaque was determined by nested polymerase chain reaction (PCR). Antichlamydial immunoglobulin G (IgG), A (IgA) and M (IgM) was examined by microimmunofluorescence and compared to the PCR result. FINDINGS: DNA of C. pneumoniae was detected in 57 (75%) of 76 atherosclerotic lesions. In most cases (74/76: 97%) a positive IgA, IgM or IgG result was seen. Seven (12%) and 54 (94%) of the 57 PCR positive plaques came from patients with a positive IgM and IgA result, respectively. There was no statistical significant difference between PCR positive and PCR negative plaques in patients with a positive or negative serological result. Clinical characteristics were similarly distributed in patients with and without infected lesions. INTERPRETATION: C. pneumoniae organisms are frequently found in the atherosclerotic lesions of patients undergoing coronary surgery for unstable angina. Neither serological results of acute or recent infection by C. pneumoniae nor clinical characteristics are useful in predicting the individual risk of harbouring C. pneumoniae in the coronary lesions of patients with unstable angina. PMID- 12767553 TI - Cardiac troponin I in aortic valve disease. AB - BACKGROUND: Plasma cardiac troponin I levels may be higher than normal in conditions other than ischemic heart disease. We aimed at measuring troponin I levels in aortic valve patients, in which increased values for left ventricular dimensions and pressure are frequently found. METHODS: Plasma levels of troponin I, creatine kinase (CK) and the MB fraction of the same enzyme were measured in a group of 25 clinically stable aortic valve patients. Echocardiographic study was performed in all patients; hemodynamic and coronary angiographic study was performed in 19 patients. Troponin I was also measured in a control population (n=305). RESULTS: The mean value for troponin I was found to be higher in aortic valve patients (0.07+/-0.02 ng/ml), when compared to controls (0.01+/-0.02 ng/ml; P<0.05). Significant correlations were found between troponin I and both creatine kinase and its MB fraction. When the 25 patients were divided into two groups, with lower (up to 0.04 ng/ml; 12 patients) and higher (0.05 ng/ml or greater; 13 patients) values for troponin I, patients with higher values were found to have greater mean left ventricular wall thickness (9.9+/-0.3 mm, n=11, vs. 12.1+/-0.3 mm, n=13) and pulmonary artery systolic pressures (36.6+/-2.5 mmHg, n=7, vs. 53.7+/-3.4 mmHg, n=9). CONCLUSIONS: We conclude that slightly raised plasma levels of cardiac troponin I are relatively common in aortic valve patients with no evidence of ischemia. Higher left ventricular wall thickness and pulmonary artery systolic pressure may be related to slightly raised troponin I plasma levels. PMID- 12767554 TI - Markers of risk in young offspring with paternal history of myocardial infarction. AB - Coronary heart disease clusters within families, but there may be several reasons for this phenomenon to occur. A possible way to elucidate this is to study biological relatives of affected individuals. The aim of our study was thus to compare a number of clinical, metabolic, clotting and immunologic factors between offspring with paternal history of premature myocardial infarction and controls and to propose a model which could safely allow to identify the high risk subgroup among them. Sixty-nine offspring of both sexes mean age 18.1 years old (cases) and thirty-two frequency matched relative to age and gender controls were studied. Cases compared to controls had significantly increased diastolic blood pressure levels (74.0+/-9.9 vs. 67.4+/-8.3 mmHg, P=0.002), leptin plasma levels (11.8+/-10.8 vs. 6.8+/-3 ng/ml, P=0.046) and fibrinogen, plasminogen, fibrin degradation products and plasminogen activator inhibitor-1 plasma levels (306.6+/ 52.5 vs. 280.6+/-28.9 mg%, P=0.03, 97.4+/-23.5 vs. 83.6+/-15 mg%, P=0.0007, 292.0+/-148.5 vs. 219.2+/-69.4 ng/ml, P=0.036, 14.7+/-5.3 vs. 8.7+/-3.1 I.U./ml, P=0.0001, respectively), while cases had significantly decreased HDL-cholesterol serum levels (45.9+/-12.5 vs. 50.5+/-8.8 mg%, P=0.03) and protein S plasma levels (89.9+/-17.5 vs. 101.3+/-13.7%, P=0.001). Our findings suggest that offspring of affected individuals may be considered as a high risk group for cardiovascular disease. PMID- 12767555 TI - Myocardial bridging of the ramus intermedius coronary artery. PMID- 12767556 TI - Reversible complete heart block due to Wegener's granulomatosis. PMID- 12767557 TI - Coronary stenting after rotational atherectomy in diffuse lesions of the small coronary artery: comparison with balloon angioplasty prior to stenting. PMID- 12767559 TI - No more unexplained trial acronyms please. Trial Acronyms: Better Obvious than Obscure (TABOO). PMID- 12767558 TI - Multivessel coronary spasm during coronary angiography: coronary vasospastic disease. PMID- 12767560 TI - Systemic embolism from a large descending aortic thrombus. AB - With the advent of transesophageal echocardiography (TEE), thrombi of the aorta are becoming increasingly recognized as possible sources of systemic emboli. This report describes a 58-year-old woman with multiple unexplained peripheral emboli. A giant thrombus of the descending aorta has been identified as the source of systemic thromboembolism. The patient refused surgery and was treated successfully with long-term anticoagulation. PMID- 12767561 TI - Accessory mitral valve tissue manifesting cerebrovascular thromboembolic event in a 34-year-old woman. AB - Accessory mitral valve tissue is an extremely rare congenital cardiac anomaly. Most of the cases reported in the medical literature were associated with left ventricular outflow tract obstruction. The majority of cases of accessory mitral valve tissue, causing left ventricular outflow tract obstruction, occur in association with other congenital cardiac anomalies. In this reported case, a patient with accessory mitral valve tissue complicated with thromboembolic cerebrovascular event is presented. The patient also had an associated idiopathic hypertrophic subaortic stenosis. PMID- 12767562 TI - Hidden hyperthyroidism in a young male patient. PMID- 12767563 TI - Left intraventricular pressure gradient induced by cardiac tamponade after thrombolysis for acute myocardial infarction. PMID- 12767564 TI - Location of the appendix in the gravid patient: a re-evaluation of the established concept. AB - OBJECTIVES: To determine the anatomical location of the appendix, as it may change during pregnancy. METHODS: In this prospective study conducted from October 1995 to March 1999, 291 women of reproductive age were evaluated for appendix location. They were divided into three groups: group A, 165 women between 37 and 40 weeks of pregnancy who underwent elective cesarean delivery; group B, 26 women between 19 and 39 weeks of gestation with acute appendicitis who underwent appendectomy; and group C (the control group), 100 non-pregnant women with acute appendicitis who underwent appendectomy. Appendix location was considered normal within 2 cm of the McBurney point; otherwise, it was considered to be a position change. RESULTS: In group A, 2 (15%) of 165 women and in group B, 6 (23%) of 26 women had a change in the position of the appendix. In the control group, 17% had change in position. There were no significant differences between group A and B compared with group C. In group B, no relation between height of fundus and point of tenderness was seen. CONCLUSION: Our study did not show that pregnancy changes the location of the appendix. PMID- 12767565 TI - Labor induction post-term with 25 micrograms vs. 50 micrograms of intravaginal misoprostol. AB - OBJECTIVES: To compare the effectiveness of 25 microg vs. 50 microg of intravaginal misoprostol for cervical ripening and labor induction beyond 41 weeks' gestation. METHODS: The study population consisted of 120 women not in active labor with a gestational age >41 weeks, singleton pregnancy with vertex presentation, reactive fetal heart rate tracing, amniotic fluid index >/=5, and Bishop score <5. Women were randomized to receive either 25 microg (n=60) or 50 microg (n=60) of intravaginal misoprostol. The dose was repeated every 4 h (maximum number of doses limited to six) until the patient exhibited three contractions in 10 min. The main outcome measure was the induction-vaginal delivery interval. RESULTS: There was no significant difference between the two groups with regard to the induction-vaginal delivery interval (685+/-201 min in the 25 microg group vs. 627+/-177 min in the 50 microg group, P=0.09). The proportion of women delivering vaginally with one dose of vaginal misoprostol was significantly greater in the 50 microg group (0/49 vs. 41/47, P<0.001). There were no differences in the rates of cesarean and operative vaginal delivery rates, or in the incidences of tachysystole and hyperstimulation syndrome in the two treatment groups. Neonatal outcomes were also similar. CONCLUSIONS: Intravaginal administration of 25 microg of misoprostol appears to be as effective as 50 microg for cervical ripening and labor induction beyond 41 weeks' gestation. PMID- 12767566 TI - Effect of lycopene on pre-eclampsia and intra-uterine growth retardation in primigravidas. AB - OBJECTIVES: To observe the effect of the antioxidant lycopene on the occurrence of pre-eclampsia and intrauterine growth retardation in primigravida women. METHODS: A total of 251 primigravida women were enrolled in this prospective, randomized controlled study in the second trimester. A total of 116 women were given oral lycopene (Group I) in a dose of 2 mg twice daily while 135 women were given a placebo (Group II) in the same dose until delivery. The criteria for recruitment included gestational age of 16-20 weeks, singleton pregnancy, absence of any medical complication and willingness on the part of the women to participate in the study. The women were followed-up until delivery for development of pre-eclampsia, mode of delivery and fetal outcome. RESULTS: The two groups were comparable in their maternal characteristics. Pre-eclampsia developed in significantly less women in the lycopene group than in the placebo group (8.6% vs. 17.7%, P=0.043 by chi-square test). Mean diastolic blood pressure was significantly higher in the placebo group (92.2+/-5.98 mmHg vs. 86.7+/-3.80 mmHg, P=0.012). Mean fetal weight was significantly higher in the lycopene group (2751.17+/-315.76 g vs. 2657+/-444.30 g, P=0.049). The incidence of intrauterine growth retardation was significantly lower in the lycopene group than in the placebo group (12% vs. 23.7%, P=0.033). CONCLUSIONS: The results of the present study suggest that the antioxidant lycopene reduces the development of pre eclampsia and intrauterine growth retardation in primigravida women. PMID- 12767567 TI - Fetal loss in threatened abortion after embryonic/fetal heart activity. AB - OBJECTIVES: To study the incidence of fetal loss in threatened abortion after detection of embryonic/fetal heart activity. METHODS: A prospective study was performed on pregnant women with clinically diagnosed threatened abortion between 6 and 14 weeks of gestation. All had a good menstrual history and the calculated gestational age using crown-rump length in the first trimester ultrasound was in agreement. Embryonic/fetal heart rate measurements were obtained by a 5 MHz vaginal probe using M-mode and real-time B mode imaging. All cases were followed up with respect to pregnancy outcomes. The data were analyzed using the SPSS computer program. RESULTS: Eighty-seven pregnant women were included in the study. There were three pregnancies (3.4%) which resulted in fetal loss before 20 weeks of gestation. In viable pregnancies, the mean embryonic/fetal heart rate increased with advancing gestational age. The individual values of embryonic/fetal heart rate for fetal losses were within the reference range. CONCLUSIONS: The incidence of fetal loss in threatened abortion after detection of embryonic/fetal heart activity was 3.4%. There was no evident pattern of bradycardia or tachycardia that signaled the incipient of viability. PMID- 12767568 TI - Prediction of perinatal asphyxia with nucleated red blood cells in cord blood of newborns. AB - OBJECTIVE: To determine normal level of nucleated red blood cells (NRBC) per 100 white blood cells (WBC) in cord blood of term non-asphyxiated newborns and to investigate variations in NRBC counts in perinatal asphyxia. METHODS: A total of 75 cases were studied. Levels of NRBC per 100 WBC in umbilical venous blood were compared between 26 asphyxiated newborns (group I) and 49 non-asphyxiated newborns (group II). Correlation with neonatal outcome was also evaluated. RESULTS: The mean (+/-S.D.) NRBC per 100 WBC level in umbilical blood of newborns in group I was 16.5+/-6.4, range 3-25; whereas that in group II was 8.6+/-7.01, range 1-26. This difference was statistically significant (P<0.001). A statistically significant negative correlation existed between NRBC level and markers of acute intrapartum asphyxia, Apgar score and umbilical arterial pH (r= 0.50, P<0.001 and r=-0.48, P<0.001, respectively). Positive correlation was demonstrated with evidence of chronic antepartum asphyxia, presence of pregnancy induced hypertension and intrauterine growth restriction (r=2.66, P=0.02). A high NRBC count in umbilical blood correlated with poor early neonatal outcome. CONCLUSIONS: The level of NRBC per 100 WBC correlates both with acute as well as chronic antepartum asphyxia. Further, it can be used as a reliable index of early neonatal outcome. PMID- 12767569 TI - Expression of insulin-like growth factor-II and insulin-like growth factor binding protein-1 in the placental basal plate from pre-eclamptic pregnancies. AB - OBJECTIVES: To determine whether expressions of insulin-like growth factor-II (IGF-II) and insulin-like growth factor binding protein-1 (IGFBP-1) are altered in pre-eclamptic placenta and to elucidate the possible relationship between their expressions and a mechanism for inadequate trophoblast invasion in pre eclampsia. METHODS: Placental tissues were obtained at cesarean delivery from five normotensive, nine mild pre-eclamptic and five severe pre-eclamptic women at 33-39 completed weeks of gestation. After total ribonucleic acid was extracted, reverse transcriptase-polymerase chain reaction was performed to determine IGF-II and IGFBP-1 mRNA expression. Product bands were quantitated by scanning densitometry and results were expressed as ratio of cytokines/beta-actin. Western blot analysis was also done to determine IGF-II and IGFBP-1 protein expression. Statistical analysis was determined by Kruskal-Wallis analysis of variance with the Scheffe multiple post-hoc test. RESULTS: The IGF-II mRNA levels of mild and severe pre-eclamptic placenta were significantly lower than that of uncomplicated placenta (P<0.005, P<0.001, respectively), with the level of severe pre-eclamptic placenta being significantly lower than that of mild pre-eclamptic placenta (P<0.05). As for the IGF-II protein expression, a significant decrease was found among the three groups (P<0.001), correlating with the IGF-II mRNA results. However, the mean IGFBP-1 mRNA levels of mild and severe pre-eclamptic placenta were significantly higher than that of uncomplicated placenta (P<0.05, P<0.005, respectively), with the level of severe pre-eclamptic placenta being significantly raised compared with that of mild pre-eclamptic placenta (P<0.05). Finally, a significant increase of IGFBP-1 protein expression was noted among the three groups (P<0.001), correlating with the IGFBP-1 mRNA results. CONCLUSIONS: This study suggests that IGF-II and IGFBP-1 might be associated with the impaired trophoblastic invasion that may lead to pathogenesis of pre-eclampsia. PMID- 12767570 TI - Retained second twins in Enugu, Nigeria. AB - OBJECTIVES: To review our experience with retained second twin in Enugu, Nigeria, over a 10-year period. METHODS: The second twin is retained when a delay of 30 min occurs after the delivery of the first twin. The case notes and records of all twin deliveries at the University of Nigeria Teaching hospital Enugu between January 1991 and December 2000 were retrieved and analyzed. RESULTS: Retained second twin occurred in 1/6 twin deliveries with a resultant perinatal mortality rate of 288.5 per 1000 deliveries which was 1.7 times that of first twin. Malpresentation (54%) and uterine atony (39%) were the principal causes of retained second twins. The place of delivery of the first twin, prolonged birth intervals, oxytocin augmentation of labor and breech extraction affected perinatal outcome. CONCLUSIONS: Early recourse to vacuum extraction and cesarean deliveries of retained second twin, effective health education and adequate supervision of primary health care facilities are advocated to reduce this preventable condition. PMID- 12767571 TI - Follicular growth, endometrial thickness, and serum estradiol levels in spontaneous and clomiphene citrate-induced cycles. AB - OBJECTIVES: To evaluate follicular growth, endometrial thickness, and serum estradiol levels in spontaneous cycles and cycles induced by clomiphene citrate (CC). METHODS: A 2-year prospective study of spontaneous cycle followed by a CC induced cycle to compare paired data. Twenty couples with unexplained infertility were recruited from the Infertility Clinic of the Jawaharlal Institute of Postgraduate Medical Education and Research. An oral dose of 150 mg of CC was administered on days 5 through 9 of the CC-induced cycle. Each woman was evaluated using transvaginal sonography and serum estradiol level estimation from day 10 of the menstrual cycle until ovulation. Results were analyzed using the paired t-test. RESULTS: The leading follicular diameter was significantly larger, endometrial thickness was significantly decreased, and serum estradiol levels were significantly higher in the follicular phase of CC-induced cycles. CONCLUSIONS: Clomiphene citrate-induced cycles showed different follicular, endometrial growth patterns, and serum estradiol levels compared with spontaneous cycles. PMID- 12767573 TI - Subcapsular hepatic hematoma. PMID- 12767572 TI - Menopause rather than estrogen modifies plasma homocysteine levels. AB - OBJECTIVES: To investigate the effects of transdermal estrogen replacement therapy (ERT) on plasma homocysteine levels in postmenopausal women who underwent total hysterectomy with bilateral oophorectomy. METHODS: In two-phase open longitudinal prospective study we compared 28 premenopausal women and 35 healthy postmenopausal patients to evaluate the effect of transdermal estrogen treatment (TTS 50 twice-weekly) on plasma homocysteine levels after 6 and 12 months of therapy. RESULTS: The study showed statistically relevant differences (P<0.05) in baseline plasma homocysteine concentration between the patients in premenopausal and in postmenopausal status. No difference in the plasma homocysteine levels was observed after 6 and 12 months of ERT on postmenopausal patients. CONCLUSIONS: Surgically postmenopausal women have higher plasma homocysteine concentrations than premenopausal women, but transdermal estrogen treatment for 12 months in postmenopausal women does not modify homocysteine levels. PMID- 12767575 TI - Consumer/provider communication research: directions for development. AB - Communication between health care providers and consumers is a critical part of effective health care delivery. Yet, the strategic use of interpersonal communication in health care delivery is most complex, multifaceted, and often problematic, necessitating careful study of the communication process in health care to increase understanding and help improve health communication practices. The National Cancer Institute (NCI) sponsored an expert symposium on consumer/provider communication research to examine progress and identify gaps in the research literature on doctor/patient communication. This paper and this special issue of Patient Education and Counseling reviews several of the key perspectives and suggestions presented at the symposium for directing future research and applications concerning consumer/provider communication. PMID- 12767576 TI - Research traditions in provider-consumer interaction: implications for cancer care. AB - The increasing importance of communication in cancer research should play a role in shaping the research agendas of those interested in investigations involving providers and consumers of cancer care. The goal of this article is to identify existing programs of research specific to provider-patient relationships and infer implications for how these research traditions can lead to important research avenues in the cancer context. Research programs reviewed in this article include (a) relational control messages and (b) patient preferences for communication style. Each section briefly reviews some noteworthy trends emanating from that research tradition and concludes with implications for future research. In the final section, two promising directions of research are proposed for consumer-provider communication. PMID- 12767577 TI - Some observations on provider-patient communication research. AB - This article presents reflections on, and suggestions for, research on provider patient communication. These include: (1) the need for more precise causal analysis, (2) considerations when doing moderator analysis, (3) the need to study values concordance between providers and patients, (4) the relative neglect of studying provider attitudes, emotions, and characteristics, (5) the potential value of studying providers' sensitivity to their patients, and (6) the importance of studying patients' impact on providers. Finally, the field is challenged to ask and answer the questions: why do we do this research and how is it being used? PMID- 12767578 TI - At the intersection of micro and macro: opportunities and challenges for physician-patient communication research. AB - The health care relationship model is undergoing dramatic change. Micro-level communication patterns yield health care relationship models (e.g. paternalism, mutual participation, consumerism). At the same time, macro-level systems appear increasingly likely to influence the nature of micro-level interaction. The intersections of health care communication micro-level and macro-level phenomena provide important venues for research and interventions. This essay identifies theoretical premises regarding the relationships between communication and health related behavior; explores three prominent and growing macro-level phenomena that observers argue likely influence the physical-patient relationship and communication therein: complementary and alternative medicine, the Internet, and direct-to-consumer advertising of prescription drugs; and offers a research agenda for exploring macro-level influences on micro-level physician-patient communication. PMID- 12767579 TI - Observations on methodological and measurement challenges in the assessment of communication during medical exchanges. AB - Reviews of medically related communication research clearly demonstrate that physicians have been studied far longer and more frequently than any other type of health care provider; nevertheless, much of what is known is limited to what is said by primarily male, White, primary care physicians during the delivery of outpatient care. Furthermore, the majority of communication studies assess medical communication largely as a physician monologue with only occasional attention to what an individual patient may say back. Limitations of the literature are obvious. Little is known about the role of a patient companion during medical visits or the consequence of patient and provider race and gender concordance (or discordance) on interpersonal dynamics. We do not know very much about relationships under stress or how patients and providers respond in those rare, but critically important events when observational techniques in the natural setting are logistically impossible or otherwise inappropriate. While it is difficult to argue with the basic tenets of ecological validity and the value of authentic settings for observational studies, the contribution of simulations and analogue studies to our understanding of medical dynamics is worthy of consideration. This essay explores methodological and design challenges related to broadening the study of medical communication through more creative and thoughtful study designs. PMID- 12767580 TI - Future directions in research on consumer-provider communication and adherence to cancer prevention and treatment. AB - The goal of this paper is to examine emerging issues in consumer-provider communication and patient adherence to cancer prevention, screening, diagnosis, treatment, and coping with survivorship. Many factors that have been shown to affect adherence can be supported or hindered by provider-patient communication, including the provider-patient relationship, patients' beliefs, social and cultural norms, family and social support, mood, and behavioral management. Six research questions are posed, and substantive and methodological recommendations are offered for empirical research on the measurement and achievement of patient adherence. PMID- 12767581 TI - Communication in context: new directions in communication research. AB - By focusing attention almost exclusively on a single encounter, researchers have adopted a rather restricted view on studying communication in health care. After all, communication does not take place in a vacuum but is influenced by the context in which it takes place. We would therefore strongly recommend to broaden the perspective of communication research. In this respect, four lines of investigation are proposed, each guided by different theories. In the first, context is determined by the goals or targets aimed at by both parties in the medical encounter. The second concerns the context of time, referring to the influence of previous and future medical encounters. The third is set up around the organizational context in which an interaction takes place and the last defines context by looking at a medical encounter as a meeting between two multifaceted parties. Studying a medical encounter in its broader context is expected to provide answers to intriguing questions such as why health care professionals do not always act in conformity with the general approved standards of high quality communication and how the factor time span can be used more effectively in the medical encounter. Eventually, a broader context view will bridge the existing gap between theory and practice. PMID- 12767582 TI - Manufactured but not imported: new directions for research in shared decision making support and skills. AB - Significant conceptual work on shared decision making has taken place but there are still significant challenges in achieving it in routine clinical practice. This paper outlines what research has identified to date that may promote shared decision making, and the further research that is required to enable continuing progress. Greater understanding of the models of decision making and instruments to identify them in practice are still required. Specifying consumer competences, developing instruments to assess these and interventions to enhance them may also be important. Clarifying all these aspects may enable those charged with training professionals to improve the content of professional development programmes. This may be particularly important in the field of cancer treatments where the stakes are high-patients usually desire much information but their desire for involvement in decision making is more variable. The consequences of getting this balance right or wrong are significant with much to be gained or lost. Continued development and evaluation of decision aids and decision explorers that use interactive technology will also be important in identifying how to progress with consumer involvement. If we can learn these lessons, then wider implementation of shared decision making or consumer involvement may become a nearer prospect. PMID- 12767583 TI - Communication and consumer decision making about cancer clinical trials. AB - Communication between patients and physicians likely mediates traditional patient and physician predispositions in determining patient outcomes, including perceptions and decision making. However, the extent to which a mediating effect occurs is unclear. The purpose of this essay is to outline the need for conceptualizing more holistic models of consumer-provider interaction that demonstrate the role of the therapeutic relationship in treatment outcomes. We focus on an important communicative context for exploring this question: the situation where patients, with the help of oncologists, are faced with making treatment choices, particularly whether to enroll in a clinical trial in response to their life-threatening cancer diagnosis. We explore the question from the perspectives of the medical provider, the patient, and the accompanying family member, in order to better frame the complex interactional dynamics occurring during the interaction. PMID- 12767584 TI - Patient centered decision making in palliative cancer treatment: a world of paradoxes. AB - Patient centered palliative cancer care would imply, first, the introduction of psychosocial endpoints when evaluating treatment and making decisions. Second, patient control would have to be enhanced by information giving and increased decision involvement. We have indicated that paradoxes exist when a patient centered approach is advocated in the context of palliative cancer care. So called patient oriented outcomes, like quality of life, once introduced seem to be disregarded by many patients themselves and survival is given a more important weight. Likewise, physicians seem to be inclined to treat patients aggressively for little benefit rather than providing supportive care. Both parties seem to prefer to do something actively to maintain a semblance of control over the disease process. Giving treatment, even if aggressive, is a way to avoid the confrontation with the little efficacy that the physician has to offer to incurable cancer patients. This mechanism is reflected in the content of conversations in palliative care. Patient centered care would imply that patient control and autonomy are enhanced. However, again paradoxically, many patients seem to want to avoid information and leave the decisions to be made by their doctors. Physicians, then, follow such wishes while paying more attention to aggressive therapy than to the notion of watchful waiting. This may help to avoid the painful confrontation with bad news. Dilemmas then remain. Patients wishing to maintain hope and avoid emotional impact of a full understanding of their prognosis may rather not be informed brusquely about prognosis or the aims of supportive therapy and forced to make an informed decision. However, by giving more aggressive, maybe even futile, treatment, and withholding supportive care patients may receive less than 'quality end-of-life care'. Therefore, information about less intrusive strategies should still be given in a cautious manner, while regarding the patient's defenses respectfully. PMID- 12767585 TI - Consumer-provider communication research with special populations. AB - As the US population becomes increasingly diverse, understanding consumer provider communication among special populations becomes of paramount importance. Cultural competence is a key element in improving communication between non minority providers and minority patients. This includes overcoming sociocultural and linguistic barriers that hinder access to care and diminish quality consumer provider communication. Of special concern is the lack of cancer prevention communication between providers and their special population patients. More research is needed to understand cancer communication needs and barriers among special populations, and to direct effective interventions to improve consumer provider communication for special populations. To this end, interactive training to improve communication skills among oncologists and medical students, increasing the availability of trained medical translators, increasing the number of health professionals from special populations, and increasing the number of special population participants in communication research are recommended. Furthermore, research should focus on identifying and overcoming cultural factors that negatively impact consumer-provider interactions. PMID- 12767586 TI - Physician-older patient communication about cancer. AB - Cancer is frequently a disease of older individuals. Communication between physicians and older patients about cancer prevention, screening, diagnosis, treatment and care is complicated by a variety of factors including patients' beliefs, perceptions, and knowledge about cancer. In addition, other older patient factors such as possible sensory deficits, cognitive impairment, functional limitations and accompaniment by significant others to the medical encounter influence communication. Physicians' attitudes about aging may also affect recommendations for cancer screening, treatment regiments and care of older cancer patients. To understand communication as a complex, multidimensional human enterprise requires knowledge of older patients' lived experience of cancer and their need for honest and compassionate care. Research findings on physician older patient communication about cancer need to be translated into medical education, training and practice to improve the care of the older cancer patient. PMID- 12767587 TI - Centering families in cancer communication research: acknowledging the impact of support, culture and process on client/provider communication in cancer management. AB - Cancer management occurs within a system of relationships. Advancing a context based view of client provider communication, we first posit that family is central to client provider communication in cancer management. This premise is followed by three additional premises that emphasize the importance of culture, disease stage and communication styles, within this family-centered perspective. Supporting research questions are offered for each premise and implications of this approach for client, family, and provider communication and for client outcomes are offered. PMID- 12767588 TI - Patient proactivity enhancing doctor-patient-family communication in cancer prevention and care among the aged. AB - This paper presents a comprehensive conceptual model of health care communication involving three key health care partners: patients, physicians, and significant family members (health significant other, HSOs). A unique feature of this model is its focus on proactive roles played by elderly patients in information gathering and communication with health care partners regarding both cancer prevention and cancer care. We outline how proactive initiatives by health care consumers and involvement of their HSOs can enhance patient outcomes (satisfaction with physician, adherence to preventive and corrective practice recommendations, and quality of life). Finally, we also note primary antecedents of health care partner communication in terms of both medical care context and patient characteristics. We hope that this testable causal model will inform future research in the field of health communication. PMID- 12767589 TI - Concordance in cancer medication management. AB - This article explores how the concept of concordance can help to identify gaps and opportunities for research on consumer-provider communication related to cancer medication management. The relationship of concordance, patient-centered care and shared decision making is examined. Research on unmet patient agendas, quality of life issues related to symptom management and tools to assist communication about patient somatic experience are discussed. The need for research on patient communication with pharmacists, nurses and other health team members beyond physicians is noted. Research implications for longitudinal, descriptive and intervention studies are offered. PMID- 12767590 TI - The interplay between education and research about patient-provider communication. AB - Attention to providers' communication skills is likely to increase, given the confluence of forces that have highlighted the importance of communication in healthcare. In the United States, interpersonal and communication skills have been explicitly identified as a priority throughout the continuum of medical education and practice. Ideally, theory and research inform teaching and assessment efforts by suggesting how communication behavior affects outcomes and by providing a conceptual framework for learning skills. This article illustrates the interplay between education and research by discussing examples of useful concepts (models of communication, issues of perceived control, and patterns of non-verbal communication) and understudied topics (physician verbalizations during patients' initial narratives, the mundane aspects of communication in healthcare, conceptual and operational definitions of empathy, and the effect of patient narratives on both patients and providers). Given the breadth and depth of experience, from screening and prevention to treatment and support, the context of cancer offers a promising laboratory for enhancing both education and research about provider-patient communication. PMID- 12767591 TI - Training practitioners to communicate effectively in cancer care: it is the relationship that counts. AB - The motivation to learn new skills that improve patient care comes from practical experience. Once motivated, trainees and practitioners alike require excellent content and process to modify approaches that improve outcomes. This paper defines content areas the authors believe are needed to improve communication between cancer patients and their practitioners. Perhaps more importantly, the educational process to achieve improved outcomes is discussed and the importance of the context in which that education occurs is stressed. The linkage between administrative behavior and practitioner behavior is described. Synchronicity between the expectations for practitioner practice and the practice environment is needed for practitioners to successfully incorporate the patient-centered practices patients are demanding. Finally, a research agenda is outlined that encourages evaluation of the model proposed. PMID- 12767592 TI - Patient communication skills training: a review with implications for cancer patients. AB - Although considerable attention has been given to physicians' communication, relatively little research has been directed to patients' communicative contributions to the medical interview. The research in patient communication skills training within the context of primary care is reviewed and implications are discussed for applying skills training in a cancer environment. PMID- 12767593 TI - Care not cure: dialogues at the transition. AB - Physicians and patients find it hard to communicate when treatment fails to cure or control cancer. Communication barriers include fear of "giving up," losing the medical team, and discussing death. The quality of physician-patient communication affects important outcomes including patient distress, coping, and quality of life, and physician burnout. Communication skills that can be taught, learned, and maintained for physicians at all levels of training, and effective educational programs have been described. Research on communication skills training should focus on the best method of delivery, the "dose-response" effect, and how to measure success of training in complex health care environments. PMID- 12767594 TI - Mediated consumer-provider communication in cancer care: the empowering potential of new technologies. AB - New communication technologies represent a potentially valuable resource for cancer care and education. With the Internet and multimedia programs (e.g. CD ROMs), health care consumers have access to a wealth of information about cancer and its treatment, can participate in online support groups, and can interact with medical experts across the globe. To be most effective, these interventions must be designed, developed, implemented, and evaluated using a sound conceptual framework that connects factors affecting utilization, the user's experience within the media environment, and post interaction outcomes. This essay presents two health communication frameworks, an expanded model of health care consumer provider communication and a three-stage model of health promotion using interactive media, to help guide future research and development of innovative technologies for cancer care and education. PMID- 12767595 TI - The association of vitamins C and K3 kills cancer cells mainly by autoschizis, a novel form of cell death. Basis for their potential use as coadjuvants in anticancer therapy. AB - Deficiency of alkaline and acid DNase is a hallmark in all non-necrotic cancer cells in animals and humans. These enzymes are reactivated at early stages of cancer cell death by vitamin C (acid DNase) and vitamin K(3) (alkaline DNase). Moreover, the coadministration of these vitamins (in a ratio of 100:1, for C and K(3), respectively) produced selective cancer cell death. Detailed morphological studies indicated that cell death is produced mainly by autoschizis, a new type of cancer cell death. Several mechanisms are involved in such a cell death induced by CK(3), they included: formation of H(2)O(2) during vitamins redox cycling, oxidative stress, DNA fragmentation, no caspase-3 activation, and cell membrane injury with progressive loss of organelle-free cytoplasm. Changes in the phosphorylation level of some critical proteins leading to inactivation of NF kappaB appear as main intracellular signal transduction pathways. The increase knowledge in the mechanisms underlying cancer cells death by CK(3) may ameliorate the techniques of their in vivo administration. The aim is to prepare the introduction of the association of vitamins C and K(3) into human clinics as a new, non-toxic adjuvant cancer therapy. PMID- 12767596 TI - Cardioselective anti-ischemic ATP-sensitive potassium channel (KATP) openers: benzopyranyl indoline and indole analogues. AB - This paper describes the design, syntheses, and biological evaluations of novel ATP-sensitive potassium channel (K(ATP)) openers, benzopyranyl indoline and indole derivatives. Among those, two enantiomers of indoline-2-carboxylic ethyl esters (14, 18) showed the best cardioprotective activities both in vitro and in vivo, while their vasorelaxation potencies were very low (concentration for 50% inhibition of vasorelaxation >30 microM). The cardioprotective effect of 14 was completely reversed by 5-hydroxydecanoate, a selective mitochondrial K(ATP) blocker, indicating its provable protective mechanism through the mitochondrial K(ATP) opening. In addition, we performed conformational analyses using 2D-NMR, X ray crystallography and molecular modeling to study the structure-activity relationships in this series of compounds. PMID- 12767597 TI - Synthesis, cytotoxic activity on MCF-7 cell line and mutagenic activity of platinum(II) complexes with 2-substituted benzimidazole ligands. AB - Four Pt(II) complexes with 2-H/or-methyl/or-aminomethylbenzimidazole or 1,2 dimethylbenzimidazole ligands as "non-leaving groups" were synthesized and their antiproliferative properties were tested against the human MCF-7 breast cancer cell line. The mutagenic potentials of the complexes were tested in Salmonella typhimurium strains TA 98 and TA 100 in the absence of S9 rat liver fraction. In general, Pt(II) complexes tested which were found to be less active than cisplatin, exhibited moderate in vitro cytotoxic activity on MCF-7 cell line. Among the complexes tested, Pt(II) complex with 2-aminomethylbenzimidazole ligand was found to be highly mutagenic in S. typhimurium TA 98 and low mutagenic in S. typhimurium TA 100. Pt(II) complex with 1,2-dimethylbenzimidazole was mutagenic only in S. typhimurium TA 98. The other two complexes were found to be non mutagen in both strains. PMID- 12767598 TI - Design, synthesis, and biological evaluation of non-peptidic ligands at the Xenopus laevis skin-melanocortin receptor. AB - Taking the tripeptide D-Trp-Arg-Leu-NH(2) as a lead for a Xenopus laevis skin melanocortin (MC) receptor antagonist, thirteen non-peptidic compounds were synthesized and biologically evaluated at Xenopus laevis melanophores. Six competitive antagonists (shown by Schild analysis) and one partial agonist were identified with moderate activity (IC(50): 5-10 microM). Tryptophanamides with aliphatic side chains were inactive whereas basic residues restored activity. Introducing an imidazole residue yielded partial agonist activity (EC50: 32 microM). Interestingly, constraining the inactive S-tryptophan-isoamylamide to a beta-carboline ring yielded an MC receptor antagonist (42). The specificity for MC receptors was tested at various G-protein coupled receptors. In conclusion, the synthesis of non-peptidic MC receptor antagonists is described which may serve as lead compounds for further studies. PMID- 12767599 TI - Structure-activity relationships of first bishydroxymethyl-substituted cage dimeric 4-aryl-1,4-dihydropyridines as HIV-1 protease inhibitors. AB - A first series of novel bishydroxymethyl-substituted cage dimeric 4-aryl-1,4 dihydropyridines 5-8 has been synthesised and evaluated as HIV-1 protease and HIV inhibitors in vitro assays. Moderate activity data of protease inhibition have been found for of the N-Boc substituted compound 8. Reduced activity for compound 6 and almost no residual activity of 5 and 7 emphasise the importance of the tert. butyl substituent for protease inhibitory activity thus supporting a discussed probable binding of the N-acyloxy substituent to the S2/S2' regions of protease. PMID- 12767600 TI - Opposite vascular activity of (R)-apomorphine and its oxidised derivatives. Endothelium-dependent vasoconstriction induced by the auto-oxidation metabolite. AB - We have synthetised a series of oxidised apomorphine derivatives (orto and para quinones 2-5), in order to analyse their vascular activity. We have performed radioligand binding assays on rat cortical membranes and functional studies on rat aortic rings. Instead the relaxant activity exhibited by (R)-apomorphine, o quinones 2, 4, show contractile activity dependent on endothelium in rat aortic rings. Compound 2, the main metabolite of (R)-apomorphine auto-oxidation, was the product which showed enhanced contractile activity by a complex mechanism related to activation of Ca(2+) channels through release and/or inhibition of endothelial factors. Moreover, this compound disrupts the endothelial function as shows the lack of response to acetylcholine observed in vessels pretreated with it. PMID- 12767601 TI - Synthesis of ibuprofen heterocyclic amides and investigation of their analgesic and toxicological properties. AB - A series of amides of ibuprofen with heteroaromatic amines was synthesized and assayed in vivo for their analgesic properties by means of writhing test in rats. When compared to parent ibuprofen some of the new amides exhibited a comparable or improved analgesic activity and a lower ulcerogenic effect. PMID- 12767602 TI - Preparation and antimicrobial behaviour of gemini fluorosurfactants. AB - The introduction of perfluorinated chains in the molecular structure of quaternary ammonium gemini surfactants have led to particularly active antimicrobial agents evaluated in this work. Connectors and spacers were studied in relation with antimicrobial activity in order to determine which molecular parameters are "critical" for biological activity. PMID- 12767604 TI - Hypervalent iodine mediated synthesis of 1-aryl/hetryl-1,2,4-triazolo[4,3-a] pyridines and 1-aryl/hetryl 5-methyl-1,2,4-triazolo[4,3-a]quinolines as antibacterial agents. AB - Oxidation of 2-pyridyl and 2-quinylhydrazones with iodobenzene diacetate (IBD) in dichloromethane yield 1-aryl/hetryl-1,2,4-trizolo-[4,3-a] pyridines (3a-f) and 1 aryl/hetryl-5-methyl-1,2,4-triazolo[4,3-a] quinolines (6a-f). Seven compounds were tested in vitro for their antibacterial activity. 1-(5'-Nitro-2-furyl)-5 methyl-1,2,4-triazolo[4,3-a]quinoline (6e) was associated with substantially higher antibacterial activity than some commercial antibiotics against Salmonella typhi at MIC i.e. 10 microg mL(-1). PMID- 12767603 TI - Synthesis and biological evaluations of pyrazolo[3,4-d]pyrimidines as cyclin dependent kinase 2 inhibitors. AB - A series of 1,4,6-trisubstituted pyrazolo[3,4-d]pyrimidines 15-19, 30-38 capable of selectively inhibiting CDK2 activity were synthesized by derivatization at C 4, C-6 and N-1 with various amines and lower alkyl groups. For above synthetic compounds, biological evaluation was carried out and structure-activity relationship was examined. In our series, 4-anilino compounds exhibited better CDK2 inhibitory activity and antitumor activity compared to 4-benzyl compounds. The compounds 33a,b having a 3-fluoroaniline group at C-4 showed comparable or superior CDK2 inhibitory activity to those of olomoucine and roscovitine as reference compounds. In general, the unsubstituted compounds (30a,b, 33a,b, 36a,b) at N-1 possessed higher potency than the substituted compounds (32a,b, 34a,b) for the CDK2 inhibitory activity. As for EGFR inhibitory activity, most compounds didnot have a significant activity. The compounds 32a,b exhibited potent cell growth inhibitory activity against human cancer cell lines, but their CDK2 inhibitory activities were slightly poorer than olomoucine. PMID- 12767605 TI - Structural requirement of isoflavonones for the inhibitory activity of interleukin-5. AB - Sophoricoside isolated from Sophora japonica is a glycoside of isoflavonone as an inhibitor of interleukin (IL)-5. To identify structural requirements of this isoflavonone for its inhibitory activity against IL-5, isoflavonones, isoflavanones, and their glycosides were prepared and their inhibitory activity was tested against IL-5. Among them, 5-benzyloxy-3-(4-hydroxyphenyl)chromen-4-one (4b, 87.9% inhibition at 50 microM, IC(50)=15.3 microM) shows the most potent activity, comparable with that of sophoricoside. The important structural requirements of these isoflavonone analogs exhibiting the inhibitory activity against IL-5 were recognized as (1) planarity of chromen-4-one ring, (2) existence of phenolic hydroxyl at 4-position of B ring, and (3) introduction of benzyloxy at 5-position, which may act as a bulky group for occupying hydrophobic pocket in putative binding site. However the glucopyranosyl moiety of sophoricoside is not an essential motif for the activity. PMID- 12767644 TI - Brugada syndrome: 1992-2002: a historical perspective. AB - An intriguing new clinical entity characterized by ST-segment elevation in the right precordial electrocardiographic leads and a high incidence of sudden death in individuals with structurally normal hearts was described by Pedro and Josep Brugada in 1992. The past decade has witnessed an exponential rise in the number of reported cases and a dramatic proliferation of papers serving to define the clinical, genetic, cellular, ionic, and molecular aspects of this disease. The purpose of this brief review is to chronicle the historical highlights that have brought us to our present understanding of Brugada syndrome. PMID- 12767645 TI - Immediate results and one-year clinical outcome after percutaneous coronary interventions in chronic total occlusions: data from a multicenter, prospective, observational study (TOAST-GISE). AB - OBJECTIVES: We sought to investigate the success rate and the acute and 12-month clinical outcome of percutaneous coronary intervention (PCI) for chronic total occlusion (CTO) in the contemporary era. BACKGROUND: The technique of PCI involving CTO has improved over time. However, limited data on acute and follow up results in patients treated with PCI on CTO in recent years are available. METHODS: Four hundred nineteen consecutive patients scheduled for PCI of CTO of > or =30 days of duration were enrolled in 29 centers; 390 CTOs were confirmed in 376 patients in an independent core laboratory. The end points were technical and procedural success, in-hospital and 12-month major adverse cardiac events (MACE) occurrence, and 12-month symptomatic status. RESULTS: Technical and procedural success was obtained in 77.2% and 73.3% of lesions, respectively. In-hospital major adverse cardiac events occurred in 5.1% of patients. Multivariate analysis identified CTO length >15 mm or not measurable, moderate to severe calcifications, duration > or =180 days, and multivessel disease as significant predictors of PCI failure. At 12 months, patients with a successful procedure experienced a lower incidence of cardiac deaths or myocardial infarction (1.05% vs. 7.23%, p = 0.005), a reduced need for coronary artery bypass surgery (2.45% vs. 15.7%, p < 0.0001), and were more frequently free of angina (88.7% vs. 75.0%, p = 0.008) compared with patients who had an unsuccessful procedure. CONCLUSIONS: Successful PCI was achieved in a high percentage of CTOs with a low incidence of complications. At one-year follow-up, patients with successful PCI of a CTO had a significantly better clinical outcome than those whose PCI was unsuccessful. PMID- 12767646 TI - Treatment of intermittent claudication with beraprost sodium, an orally active prostaglandin I2 analogue: a double-blinded, randomized, controlled trial. AB - OBJECTIVES: In the current study, we hypothesized that beraprost would: 1) improve treadmill exercise performance and quality of life; and 2) decrease rates of ischemic events in patients with intermittent claudication. BACKGROUND: Previous trials with beraprost sodium, an orally active prostaglandin I(2) analogue, in the treatment of claudication in patients with peripheral arterial disease (PAD) have been inconsistent. METHODS: Patients with intermittent claudication (n = 897) were randomized to receive either 40 microg three times a day of beraprost with meals (n = 385) or placebo (n = 377) in a double-blinded manner for one year. The primary efficacy parameter was treadmill-measured maximum walking distance, as assessed at three and six months after randomization. Secondary efficacy parameters included treadmill-measured pain free walking distance and change in quality of life. RESULTS: There was no significant improvement in maximum walking distance in the beraprost group (16.7%) as compared with the placebo group (14.6%, p = NS). Administration of beraprost did not improve the pain-free walking distance (p = NS between treatment groups), and there was no improvement in the quality-of-life measures between the treatment groups. The incidence of critical cardiovascular events was 7.3% in the beraprost group and 11.4% in the placebo group (p = NS). There was a significant reduction in the combination of cardiovascular death and myocardial infarction in the beraprost group (p = 0.01). CONCLUSIONS: Despite previous investigations suggesting efficacy, these results indicate that beraprost is not an effective treatment to improve symptoms of intermittent claudication in patients with PAD. The potential benefit of beraprost on critical cardiovascular events would require confirmation in a larger prospective investigation. PMID- 12767647 TI - Beraprost for the treatment of intermittent claudication. PMID- 12767648 TI - Randomized trial of rate-control versus rhythm-control in persistent atrial fibrillation: the Strategies of Treatment of Atrial Fibrillation (STAF) study. AB - OBJECTIVES: This study was designed to compare two treatment strategies in patients with atrial fibrillation(AF): rhythm-control (restoration and maintenance of sinus rhythm) and rate-control (pharmacologic or invasive rate control and anticoagulation). BACKGROUND: Atrial fibrillation is the most common arrhythmia. It is unclear whether a strategy of rhythm- or rate-control is better in terms of mortality, morbidity, and quality of life. METHODS: The Strategies of Treatment of Atrial Fibrillation (STAF) multicenter pilot trial randomized 200 patients (100 per group) with persistent AF to rhythm- or rate-control. The combined primary end point was a combination of death, cardiopulmonary resuscitation, cerebrovascular event, and systemic embolism. RESULTS: After 19.6 +/- 8.9 months (range 0 to 36 months) there was no difference in the primary end point between rhythm-control (9/100; 5.54%/year) and rate-control (10/100; 6.09%/year; p = 0.99). The percentage of patients in sinus rhythm in the rhythm control group after up to four cardioversions during the follow-up period (rate control group) was 23% (0%) at 36 months. Eighteen primary end points occurred in atrial fibrillation; only one occurred in sinus rhythm (p = 0.049). CONCLUSIONS: The STAF pilot study showed no differences between the two treatment strategies in all end points except hospitalizations. These data suggest that there was no benefit in attempting rhythm-control in these patients with a high risk of arrhythmia recurrence. It remains unclear whether the results in the rhythm control group would have been better if sinus rhythm had been maintained in a higher proportion of patients, as all but one end point occurred during AF. PMID- 12767649 TI - The Australian Intervention Randomized Control of Rate in Atrial Fibrillation Trial (AIRCRAFT). AB - OBJECTIVES: The Australian Intervention Randomized Control of Rate in Atrial Fibrillation Trial was a multicenter trial of atrioventricular junction ablation and pacing (AVJAP) compared with pharmacologic ventricular rate control (medication [MED]) in patients with mild to moderately symptomatic permanent atrial fibrillation (AF). BACKGROUND: There have been very few prospective randomized trials, undertaken in highly symptomatic patients, comparing AVJAP with pharmacologic methods of ventricular rate control for patients with permanent AF. METHODS: There were 99 patients (70 men, mean age 68 +/- 8.6 years) at five centers. Forty-nine patients were randomized to AVJAP while 50 patients were randomized to pharmacologic control. The primary end point was cardiac function measured by echocardiography and exercise tolerance. The secondary end points were ventricular rate control, evaluated by 24-h ambulatory electrocardiographic monitoring, and quality of life. Data were collected at randomization and then at one month, six months, and 12 months post randomization. RESULTS: At 12 months follow-up there was no significant difference in left ventricular ejection fraction (AVJAP: 54 +/- 17%; MED: 61 +/- 13% [p = ns]) or exercise duration on treadmill testing (AVJAP: 4.1 +/- 2 min; MED: 4.6 +/- 2 min [p = ns]); however, the peak ventricular rate was lower in the AVJAP group during exercise (112 +/- 17 beats/min vs. 153 +/- 36 beats/min, p < 0.05) and activities of daily life (117 +/- 16 beats/min vs. 152 +/- 37 beats/min, p < 0.05). The CAST quality-of-life questionnaire revealed that patients in the AVJAP group had fewer symptoms at six months (p = 0.003) and at 12 months (p = 0.004). The observed relative risk reduction in symptoms at 12 months was 18%. Global subjective semiquantitative measurement of quality of life using the "ladder of life" revealed that the AVJAP group reported a 6% better quality of life at six months (p = 0.011). CONCLUSIONS: In this trial, AVJAP for patients with mild to moderately symptomatic permanent AF did not worsen cardiac function during long-term follow-up, and quality of life was improved. PMID- 12767650 TI - Rhythm management in atrial fibrillation: less is more. PMID- 12767651 TI - Amiodarone versus implantable cardioverter-defibrillator:randomized trial in patients with nonischemic dilated cardiomyopathy and asymptomatic nonsustained ventricular tachycardia--AMIOVIRT. AB - OBJECTIVES: The purpose of this multicenter randomized trial was to compare total mortality during therapy with amiodarone or an implantable cardioverter defibrillator (ICD) in patients with nonischemic dilated cardiomyopathy (NIDCM) and nonsustained ventricular tachycardia (NSVT). BACKGROUND: Whether an ICD reduces mortality more than amiodarone in patients with NIDCM and NSVT is unknown. METHODS: One hundred three patients with NIDCM, left ventricular ejection fraction < or =0.35, and asymptomatic NSVT were randomized to receive either amiodarone or an ICD. The primary end point was total mortality. Secondary end points included arrhythmia-free survival, quality of life, and costs. RESULTS: The study was stopped when the prospective stopping rule for futility was reached. The percent of patients surviving at one year (90% vs. 96%) and three years (88% vs. 87%) in the amiodarone and ICD groups, respectively, were not statistically different (p = 0.8). Quality of life was also similar with each therapy (p = NS). There was a trend with amiodarone, as compared to the ICD, towards improved arrhythmia-free survival (p = 0.1) and lower costs during the first year of therapy ($8,879 US dollars vs. $22,039 US dollars, p = 0.1). CONCLUSIONS: Mortality and quality of life in patients with NIDCM and NSVT treated with amiodarone or an ICD are not statistically different. There is a trend towards a more beneficial cost profile and improved arrhythmia-free survival with amiodarone therapy. PMID- 12767652 TI - Say no to primary prophylaxis with implantable cardioverter-defibrillators in asymptomatic nonischemic dilated cardiomyopathy? PMID- 12767653 TI - Concomitant recovery of atrial mechanical and endocrine function after cardioversion in patients with persistent atrial fibrillation. AB - OBJECTIVES: The purpose of this study was to evaluate left atrial mechanical function recovery and plasma atrial natriuretic peptide (ANP) release following successful cardioversion of persistent atrial fibrillation (AF). BACKGROUND: Atrial fibrillation is characterized by functional deterioration, loss of atrial contraction, and elevation of plasma ANP levels. The response of ANP release toward atrial mechanical function after cardioversion of AF has not been fully examined. METHODS: We examined 29 patients with successfully cardioverted persistent AF in whom sinus rhythm was maintained for at least 30 days after cardioversion. We assessed mechanical function of the left atrium at 24 h and 7 and 30 days after cardioversion and evaluated plasma ANP level at the same time. Atrial mechanical function was assessed during echocardiographic examination by means of the peak velocity of the transmitral A-wave, early transmitral to atrial flow velocity ratio, and atrial filling fraction (AFF). The plasma ANP level was determined by the radioimmunoassay method. RESULTS: Plasma ANP levels were significantly reduced from 59.4 +/- 16.6 pg/ml to 31.1 +/- 9.2 pg/ml at 24 h after successful cardioversion. Within 30 days, we noted progressive improvement of atrial systolic function (increase in AFF from 21% to 31%, p < 0.05). At the same time, plasma ANP levels gradually increased from 31.1 +/- 9.2 pg/ml at 24 h to 36.9 +/- 12.8 pg/ml on day 30 following cardioversion (p < 0.05). CONCLUSIONS: Plasma ANP levels significantly decreased in patients with persistent AF after successful cardioversion. In the 30 days after cardioversion, gradual elevation of plasma ANP concentration was observed concomitantly with an increase of AFF. Plasma ANP release after successful cardioversion of persistent AF might be due to recovery of atrial mechanical function. PMID- 12767654 TI - Catheter-based autologous bone marrow myocardial injection in no-option patients with advanced coronary artery disease: a feasibility study. AB - OBJECTIVES: We conducted a pilot study to evaluate the feasibility of transendocardial delivery of autologous bone marrow (ABM) strategy in patients with severe symptomatic chronic myocardial ischemia not amenable to conventional revascularization. BACKGROUND: Transendocardial injection of ABM cells appears to enhance perfusion of ischemic porcine myocardium. METHODS: Ten patients underwent transendocardial injection of freshly aspirated and filtered unfractionated ABM using left ventricular electromechanical guidance. Twelve injections of 0.2 ml each were successfully delivered into ischemic noninfarcted myocardium pre identified by single-photon emission computed tomography perfusion imaging. RESULTS: Autologous bone marrow injection was successful in all patients and was associated with no serious adverse effects; in particular, there was no arrhythmia, evidence of infection, myocardial inflammation, or increased scar formation. Two patients were readmitted for recurrent chest pain. At three months, Canadian Cardiovascular Society angina score significantly improved (3.1 +/- 0.3 vs. 2.0 +/- 0.94, p = 0.001), as well as stress-induced ischemia occurring within the injected territories (2.1 +/- 0.8 vs. 1.6 +/- 0.8, p < 0.001). Treadmill exercise duration, available in nine patients, increased, but the change was not significant (391 +/- 155 vs. 485 +/- 198, p = 0.11). CONCLUSIONS: This study provides preliminary clinical data indicating feasibility of catheter-based transendocardial delivery of ABM to ischemic myocardium. PMID- 12767655 TI - Intravascular brachytherapy for native coronary ostial in-stent restenotic lesions. AB - OBJECTIVES: We analyzed the effects of vascular brachytherapy (VBT) on ostial in stent restenosis (ISR). BACKGROUND: In-stent restenosis has a high recurrence rate after percutaneous reintervention. The recurrence rate of ostial ISR lesions and the impact of VBT remain unknown. METHODS: We evaluated 133 patients with native coronary ostial ISR from a pooled database of 990 patients enrolled in randomized VBT trials. Independent quantitative angiography was performed at baseline and follow-up in 45 gamma, 27 beta, and 61 placebo patients. RESULTS: Binary restenosis was significantly higher in placebo than radiated patients (75.4% vs. 17.8% in gamma vs. 22.2% in beta, p < 0.0001). The treatment effect of both gamma (odds ratio [OR] 0.06; 95% confidence interval [CI] 0.02 to 0.17) and beta VBT (OR 0.10; 95% CI 0.03 to 0.31) was maintained after controlling for differences in baseline lesion length. Proximal and distal radiation edge restenosis rates were similar among the groups. Vascular brachytherapy of true aorto-ostial lesions (n = 34) was similarly beneficial: restenosis rates of placebo versus gamma or beta patients of 83.3% versus 6.7% versus 28.6%, p = 0.0002. CONCLUSIONS: Conventional treatment of ostial ISR is associated with a recurrence rate of over 75%. Vascular brachytherapy with either gamma or beta sources results in significant and similar reductions in restenosis compared with placebo. Similar benefits after VBT prevail in true aorto-ostial lesions. PMID- 12767656 TI - Health-related quality of life after percutaneous coronary intervention versus coronary bypass surgery in high-risk patients with medically refractory ischemia. AB - OBJECTIVES: We compared six-month health-related quality of life (HRQL) for high risk patients with medically refractory ischemia randomized to percutaneous coronary intervention (PCI) versus coronary artery bypass graft (CABG) surgery. BACKGROUND: Mortality rates after PCI and CABG surgery are similar. Therefore, differences in HRQL outcomes may help in the selection of a revascularization procedure. METHODS: Patients were enrolled in a Veterans Affairs multicenter randomized trial comparing PCI versus CABG for patients with medically refractory ischemia and one or more risk factors for adverse outcome; 389 of 423 patients (92%) alive six months after randomization completed an Short Form-36 (SF-36) health status survey. Primary outcomes were the Physical Component Summary (PCS) and Mental Component Summary (MCS) scores from the SF-36. Multivariable analyses were used to evaluate whether PCI or CABG surgery was associated with better PCS or MCS scores after adjusting for over 20 baseline variables. RESULTS: There were no significant differences in either PCS scores (38.7 vs. 37.3 for PCI and CABG, respectively; p = 0.23) or MCS scores (45.5 vs. 46.1, p = 0.58) between the treatment arms. In multivariable models, there remained no difference in HRQL for post-PCI versus post-CABG patients (for PCS, absolute difference = 0.56 +/- standard error of 1.14, p = 0.63; for MCS, absolute difference = -1.23 +/- 1.12, p = 0.27). We had 97% power to detect a four-point difference in scores, where four to seven points is a clinically important difference. CONCLUSIONS: High-risk patients with medically refractory ischemia randomized to PCI versus CABG surgery have equivalent six-month HRQL. Therefore, HRQL concerns should not drive decision-making regarding selection of a revascularization procedure for these patients. PMID- 12767657 TI - Repeated intravascular ultrasound imaging in cardiac transplant recipients does not accelerate transplant coronary artery disease. AB - OBJECTIVES: This study was designed to examine the impact of repeated intravascular ultrasound (IVUS) examinations on transplant coronary artery disease (CAD). BACKGROUND: Serial IVUS is the most accurate method for early detection and surveillance of transplant CAD. However, the long-term safety of serial IVUS exams is not well described. Accordingly, we examined the impact of repeated IVUS examinations on transplant CAD. METHODS: We examined 226 transplant recipients who underwent one or more IVUS examinations and coronary angiography at least one year after the last IVUS exam. The coronary angiograms were analyzed using quantitative coronary angiography. Vessel diameters, frequency, and severity of stenoses in IVUS-imaged and nonimaged coronary arteries were compared. In a subgroup analysis of 31 patients, angiographic lumen diameters were measured at baseline (within eight weeks of transplantation) and during follow-up (after two, three, or four IVUS studies). RESULTS: In the 226 patients, 548 coronary arteries were previously imaged by IVUS and 130 arteries were not imaged by IVUS. On subsequent angiograms, stenoses were observed in 16.2% (21/130) of nonimaged arteries and 19.5% (107/548) of imaged arteries (p = 0.38). The arterial diameters of nonimaged and imaged arteries were not significantly different (p = 0.07), regardless of the number of IVUS exams and duration of follow-up. Subgroup analysis revealed a significant decrease in vessel lumen diameter over time in nonimaged as well as imaged arteries. The magnitude of the diameter decrease was not significantly different between the two groups. CONCLUSIONS: Repeated IVUS examinations following heart transplantation do not result in angiographically evident acceleration of transplant CAD. Therefore, serial IVUS imaging is a safe method for the detection and surveillance of transplant CAD. PMID- 12767658 TI - Effect of supplemental phytonutrients on impairment of the flow-mediated brachial artery vasoactivity after a single high-fat meal. AB - OBJECTIVES: Our objective was to determine if long-term daily administration of phytonutrient supplements can prevent the immediate adverse impact of a high-fat meal and increase the production of nitric oxide. BACKGROUND: Ingestion of a high fat meal impairs flow-mediated vasodilation of the brachial artery for at least 4 h; however, co-ingestion of vitamin antioxidants or a green salad has been shown to prevent this effect. METHODS: Flow-mediated brachial artery reactivity test (BART) both before and 3 h after a 900 calorie 50 g fat meal was evaluated in 38 healthy volunteers (age 36.4 +/- 10.1 years). Subjects were randomized to four weeks of daily supplementation with a powdered fruit vegetable juice concentrate (Juice Plus [JP]) along with a complex supplement providing nutritional antioxidants and various herbal extracts (Vineyard [V]), JP alone, or a matching placebo. At three and four weeks, BART was repeated both before and after the high-fat meal. Serum nitrate/nitrite concentrations were measured at baseline and at four weeks. RESULTS: Four weeks of the JP-V combination blunted the detrimental effect of the high-fat meal (-47.5 +/- 23.4% at baseline vs. -1.7 +/- 9.7% at four weeks [p < 0.05]). Four weeks of JP alone had a similar beneficial effect (-45.1 +/- 19.7% at baseline vs. -16.6 +/- 10.3% at four weeks [p < 0.05]), whereas there was no substantial effect of the placebo. In the subjects treated with supplements, concentrations of serum nitrate/nitrite increased from 78 +/- 39 to 114 +/- 62 microm/l (p < 0.02). CONCLUSIONS: Daily ingestion of modest amounts of a fruit/vegetable juice concentrate with or without adjunctive phytonutrient supplementation can reduce the immediate adverse impact of high-fat meals on flow-mediated vasoactivity and increase nitrate/nitrite blood concentration. PMID- 12767659 TI - High-fat diets and cardiovascular disease: are nutritional supplements useful? PMID- 12767660 TI - Pulse pressure and endothelial dysfunction in never-treated hypertensive patients. AB - OBJECTIVES: This study sought to investigate whether pulse pressure (PP) is associated with endothelium-dependent vasodilation in a group of never-treated hypertensives. BACKGROUND: Pulse pressure represents a well-established independent predictor for cardiovascular morbidity and mortality. Forearm endothelial dysfunction, defined as impaired vasodilating response to acetylcholine (ACh), may be associated with several cardiovascular risk factors. Recently, the prognostic value of coronary and forearm endothelial dysfunction has been demonstrated. METHODS: All patients underwent measurement of blood pressure (BP) both clinically and in an ambulatory setting. Endothelium-dependent and -independent vasodilation was investigated by strain-gauge plethysmography in 262 hypertensive patients (age 30 to 55 years) during intra-arterial infusion of increasing doses of ACh and sodium nitroprusside. RESULTS: We observed that systolic BP rather than diastolic BP significantly induces the PP increase. Linear regression analysis revealed a significant inverse correlation between ACh stimulated forearm blood flow (FBF) and age, body mass index, clinic and monitored systolic BP, and clinic and monitored PP. However, stepwise multivariate analysis showed that monitored PP was the strongest independent predictor of ACh-stimulated FBF, accounting for 33.6% of the variation. After adjustment for other covariates, ACh-stimulated FBF decreases by 8.7% for each mm Hg increment in monitored PP. CONCLUSIONS: Our data indicate that monitored PP is inversely correlated with ACh-stimulated vasodilation. It is possible to hypothesize that elevation in PP reduces FBF by increasing oxidative stress and reducing production of nitric oxide caused by reduced shear stress. In addition, the present findings demonstrate the accuracy of ambulatory BP as a prognostic predictor of hypertension-associated endothelial dysfunction. PMID- 12767661 TI - Endothelial function under pressure. PMID- 12767662 TI - Enhanced external counterpulsation improves endothelial function in patients with symptomatic coronary artery disease. AB - OBJECTIVES: The goal of this study was to examine the effect of enhanced external counterpulsation (EECP) on endothelial function. BACKGROUND: Enhanced external counterpulsation improves symptoms and exercise tolerance in patients with symptomatic coronary artery disease (CAD). However, the exact mechanisms by which this technique exerts its clinical benefit are unclear. METHODS: Reactive hyperemia-peripheral arterial tonometry (RH-PAT), a noninvasive method to assess peripheral endothelial function by measuring reactive hyperemic response in the finger, was performed in 23 patients with refractory angina undergoing a 35-h course of EECP. In each patient RH-PAT measurements were performed before and after the first, at midcourse, and the last EECP session. In addition, RH-PAT response was assessed one month after completion of EECP therapy; RH-PAT index, a measure of reactive hyperemia, was calculated as the ratio of the digital pulse volume during reactive hyperemia divided by that at rest. RESULTS: Enhanced external counterpulsation led to symptomatic improvement (>/=1 Canadian Cardiovascular Society class) in 17 (74%) patients; EECP was associated with a significant immediate increase in average RH-PAT index after each treatment (p < 0.05). In addition, average RH-PAT index at one-month follow-up was significantly higher than that before EECP therapy (p < 0.05). When patients were divided by their clinical response, RH-PAT index at one-month follow-up increased only in those patients who experienced clinical benefit. CONCLUSIONS: Enhanced external counterpulsation enhances peripheral endothelial function with beneficial effects persisting at one-month follow-up in patients with a positive clinical response. This suggests that improvement in endothelial function may contribute to the clinical benefit of EECP in patients with symptomatic CAD. PMID- 12767663 TI - Predictive value of noninvasively determined endothelial dysfunction for long term cardiovascular events in patients with peripheral vascular disease. AB - OBJECTIVES: The goal of this study was to prospectively examine the long-term predictive value of brachial-artery endothelial dysfunction for future cardiovascular events. BACKGROUND: Brachial-artery endothelial function is impaired in individuals with atherosclerosis and coronary risk factors. The prospective relation between endothelial function determined by brachial-artery ultrasound and long-term cardiovascular risk is unknown. METHODS: We examined brachial-artery endothelial function using ultrasound in 199 patients with peripheral arterial disease before elective vascular surgery. Patients were prospectively followed with an average follow-up of 1.2 years after surgery. RESULTS: Thirty-five patients had an event during follow-up, including cardiac death (5 patients), myocardial infarction (17 patients), unstable angina (10 patients), or stroke (3 patients). Preoperative endothelium-dependent flow mediated dilation (FMD) was significantly lower in patients with an event (4.4 +/ 2.8%) compared with those without an event (7.0 +/- 4.9%, p < 0.001), whereas endothelium-independent vasodilation to nitroglycerin was similar in both groups. In a Cox proportional-hazards model, independent predictors of events included age (p = 0.003), more invasive surgery (surgery other than carotid endarterectomy, p = 0.02), and impaired brachial-artery endothelial function (p = 0.002). Risk was approximately nine-fold higher in patients with FMD <8.1% (lower two tertiles) compared with those in the upper tertile (odds ratio 9.5; 95% confidence interval 2.3 to 40). CONCLUSIONS: Impaired brachial-artery endothelial function independently predicts long-term cardiovascular events in patients with peripheral arterial disease. The findings suggest that noninvasive assessment of endothelial function using brachial-artery FMD may serve as a surrogate end point for cardiovascular risk. PMID- 12767664 TI - Hypertrophic cardiomyopathy due to sarcomeric gene mutations is characterized by impaired energy metabolism irrespective of the degree of hypertrophy. AB - OBJECTIVES: We investigated cardiac energetics in subjects with mutations in three different familial hypertrophic cardiomyopathy (HCM) disease genes, some of whom were nonpenetrant carriers without hypertrophy, using phosphorus-31 magnetic resonance spectroscopy. BACKGROUND: Familial hypertrophic cardiomyopathy is caused by mutations in sarcomeric protein genes. The mechanism by which these mutant proteins cause disease is uncertain. A defect of myocyte contractility had been proposed, but in vitro studies of force generation have subsequently shown opposing results in different classes of mutation. An alternative hypothesis of "energy compromise" resulting from inefficient utilization of adenosine triphosphate (ATP) has been suggested, but in vivo data in humans with genotyped HCM are lacking. METHODS: The cardiac phosphocreatine (PCr) to ATP ratio was determined at rest in 31 patients harboring mutations in the genes for either beta-myosin heavy chain, cardiac troponin T, or myosin-binding protein C, and in 24 controls. Transthoracic echocardiography was used to measure left ventricular (LV) dimensions and maximal wall thickness. RESULTS: The PCr/ATP was reduced in the HCM subjects by 30% relative to controls (1.70 +/- 0.43 vs. 2.44 +/- 0.30; p < 0.001), and the reduction was of a similar magnitude in all three disease-gene groups. The PCr/ATP was equally reduced in subjects with (n = 24) and without (n = 7) LV hypertrophy. CONCLUSIONS: Our data provide evidence of a bioenergetic deficit in genotype-confirmed HCM, which is present to a similar degree in three disease-gene groups. The presence of energetic abnormalities, even in those without hypertrophy, supports a proposed link between altered cardiac energetics and development of the disease phenotype. PMID- 12767665 TI - Can an energy-deficient heart grow bigger and stronger? PMID- 12767666 TI - A homoplasmic mitochondrial transfer ribonucleic acid mutation as a cause of maternally inherited hypertrophic cardiomyopathy. AB - OBJECTIVES: The purpose of this study was to understand the clinical and molecular features of familial hypertrophic cardiomyopathy (HCM) in which a mitochondrial abnormality was strongly suspected. BACKGROUND: Defects of the mitochondrial genome are responsible for a heterogeneous group of clinical disorders, including cardiomyopathy. The majority of pathogenic mutations are heteroplasmic, with mutated and wild-type mitochondrial deoxyribonucleic acid (mtDNA) coexisting within the same cell. Homoplasmic mutations (present in every copy of the genome within the cell) present a difficult challenge in terms of diagnosis and assigning pathogenicity, as human mtDNA is highly polymorphic. METHODS: A detailed clinical, histochemical, biochemical, and molecular genetic analysis was performed on two families with HCM to investigate the underlying mitochondrial defect. RESULTS: Cardiac tissue from an affected child in the presenting family exhibited severe deficiencies of mitochondrial respiratory chain enzymes, whereas histochemical and biochemical studies of the skeletal muscle were normal. Mitochondrial DNA sequencing revealed an A4300G transition in the mitochondrial transfer ribonucleic acid (tRNA)(Ile) gene, which was shown to be homoplasmic by polymerase chain reaction/restriction fragment length polymorphism analysis in all samples from affected individuals and other maternal relatives. In a second family, previously reported as heteroplasmic for this base substitution, the mutation has subsequently been shown to be homoplasmic. The pathogenic role for this mutation was confirmed by high-resolution Northern blot analysis of heart tissue from both families, revealing very low steady-state levels of the mature mitochondrial tRNA(Ile). CONCLUSIONS: This report documents, for the first time, that a homoplasmic mitochondrial tRNA mutation may cause maternally inherited HCM. It highlights the significant contribution that homoplasmic mitochondrial tRNA substitutions may play in the development of cardiac disease. A restriction of the biochemical defect to the affected tissue has important implications for the screening of patients with cardiomyopathy for mitochondrial disease. PMID- 12767667 TI - Clinical assessment identifies hemodynamic profiles that predict outcomes in patients admitted with heart failure. AB - OBJECTIVES: This study was designed to determine the relevance of a proposed classification for advanced heart failure (HF). Profiles based on clinical assessment of congestion and perfusion at the time of hospitalization were compared with subsequent outcomes. BACKGROUND: Optimal design of therapy and trials for advanced HF remains limited by the lack of simple clinical profiles to characterize patients. METHODS: Prospective analysis was performed for 452 patients admitted to the cardiomyopathy service at the Brigham and Women's Hospital with a diagnosis of HF. Patients were classified by clinical assessment into four profiles: profile A, patients with no evidence of congestion or hypoperfusion (dry-warm, n = 123); profile B, congestion with adequate perfusion (wet-warm, n = 222); profile C, congestion and hypoperfusion (wet-cold, n = 91); and profile L, hypoperfusion without congestion (dry-cold, n = 16). Other standard predictors of outcome were included and patients were followed for the end points of death (n = 117) and death or urgent transplantation (n = 137) at one year. RESULTS: Survival analysis showed that clinical profiles predict outcomes in HF. Profiles B and C increase the risk of death plus urgent transplantation by univariate (hazard ratio [HR] 1.83, p = 0.02) and multivariate analyses (HR 2.48, p = 0.003). Moreover, clinical profiles add prognostic information even when limited to patients with New York Heart Association (NYHA) class III/IV symptoms (profile B: HR 2.23, p = 0.026; profile C: HR 2.73, p = 0.009). CONCLUSIONS: Simple clinical assessment can be used to define profiles in patients admitted with HF. These profiles predict outcomes and may be used to guide therapy and identify populations for future investigation. PMID- 12767668 TI - Serum levels of carbohydrate antigen 125 in patients with chronic heart failure: relation to clinical severity, hemodynamic and Doppler echocardiographic abnormalities, and short-term prognosis. AB - OBJECTIVES: The aim of this study was to evaluate the serum levels of carbohydrate antigen 125 (CA125) in patients with congestive heart failure (CHF). BACKGROUND: CA125 is a glycoprotein produced by serosal epithelium, found to be increased in ovarian cancer. METHODS: Serum levels of CA125 were obtained in 286 patients (122 males and 164 females; age 69 +/- 13 years) with CHF (left ventricular ejection fraction 30 +/- 11%). A non-invasive evaluation was obtained by Doppler echocardiography; right heart catheterization was performed in 88 patients. An attempt to adjust medical therapy to maximally tolerated doses was done, and CA125 was repeated after 18 days (range 7 to 40) in 80 patients. The mean follow-up duration was 6 +/- 3 months in 240 patients. RESULTS: The mean value of CA125 was 68 +/- 83 U/ml (range 3 to 537): 71 +/- 85 in men and 56 +/- 64 U/ml in women (p = NS). CA125 above the normal value (<35 U/ml) was found in 152 (53%) of 286 patients; it was higher in patients with advanced New York Heart Association (NYHA) functional class (n = 140 in class I/II: 15 +/- 9 U/ml; n = 63 in class III: 57 +/- 18 U/ml; n = 83 in class IV: 167 +/- 94 U/ml; p < 0.005). CA125 was related to the deceleration time of early filling on transmitral Doppler (r = -0.63, p < 0.05) and to pulmonary artery wedge pressure (r = 0.66, p < 0.05) and right atrial pressure (r = 0.69, p < 0.05). During 6 +/- 3 months of follow-up, a combined end point of mortality and CHF hospitalization was observed in 16 of 127 patients with CA125 <35 U/ml, compared with 70 of 113 patients with CA125 >35 U/ml (p < 0.01). After medical treatment optimization, NYHA class decreased by more than one grade in 56 of 80 patients and was unchanged or increased in 24 patients: CA125 decreased from 125 +/- 98 to 53 +/- 61 U/ml (p < 0.001) in the former and changed from 130 +/- 81 to 153 +/- 61 U/ml (p = NS) in the latter. CONCLUSIONS: Our data suggest that CA125 is related to CHF severity and short-term prognosis. Furthermore, fluctuations of CA125 serum levels over time may reflect changes induced by therapy. Therefore, measurements of CA 125 serum levels might be proposed for the serial assessment of CHF patients. PMID- 12767670 TI - Cyclooxygenase-2 inhibitors and atherosclerosis. PMID- 12767669 TI - Effects of MF-tricyclic, a selective cyclooxygenase-2 inhibitor, on atherosclerosis progression and susceptibility to cytomegalovirus replication in apolipoprotein-E knockout mice. AB - OBJECTIVES: We examined whether selective cyclooxygenase-2 (COX-2) inhibition in apolipoprotein-E (apoE) deficient mice reduces cytomegalovirus (CMV) replication, and determined whether COX-2 anti-inflammatory activity leads to decreased atherosclerosis. BACKGROUND: Evidence suggests that CMV infection contributes to atherosclerosis and that this occurs in part through inflammatory mechanisms. Cyclooxygenase-2 inhibitors are potent anti-inflammatory agents. They also inhibit CMV replication in vitro. METHODS: The apoE deficient mice were either treated or not treated with a selective COX-2 inhibitor, and either infected or not infected with CMV. Viral deoxyribonucleic acid load in salivary glands was determined by quantitative polymerase chain reaction. Atherosclerotic lesion analysis was performed by standard methods. RESULTS: In vivo COX-2 inhibition, unexpectedly increased viral load: in the CMV-infected animals viral load was 2.58 +/- 1.0 in the nontreated group, 4.74 +/- 1.38 in the group treated with 12 mg/kg/day MF-tricyclic, and 6.51 +/- 1.64 in the group treated with 24 mg/kg/day MF-tricyclic (p trend = 0.050). This increased viral load was paralleled by increased anti-CMV antibody titers. Most surprisingly, COX-2 inhibition significantly increased early atherosclerotic lesion area, independent of viral infection. CONCLUSIONS: Our study demonstrates that selective inhibition of COX-2 in vivo increases viral load. The finding that inhibition of COX-2 increases atherosclerosis development in apoE deficient mice suggests, unexpectedly, that this enzyme exerts antiatherosclerosis activity, at least in this model. PMID- 12767671 TI - Assessment of right ventricular perfusion after right coronary artery occlusion by myocardial contrast echocardiography. AB - OBJECTIVES: The purpose of this study was to examine the ability of myocardial contrast echocardiography (MCE) to assess right ventricular (RV) perfusion. BACKGROUND: Although MCE can readily assess left ventricular perfusion abnormalities, there are no data regarding the ability to assess RV perfusion abnormalities. METHODS: The right coronary artery (RCA) was occluded in 10 open chest dogs. Myocardial contrast echocardiography was performed with 0.27 g/min Levovist infusion by harmonic power Doppler with electrocardiographically gated intermittent triggered imaging at pulsing intervals ranging from 1:1 to 1:20 at baseline and 90 min after RCA occlusion. Video-intensity of the RV wall was plotted against pulsing intervals and was fitted to an exponential function: y = A(1-exp(-bt)), where A is the plateau video-intensity and b is the rate of video intensity rise. Myocardial contrast echocardiography and microsphere-derived myocardial blood flow (MBF) measurements were performed at baseline and 90 min after RCA occlusion. RESULTS: Because the severity of RV perfusion abnormalities assessed by MBF varied during RCA occlusion, diverse grades of patchy opacification defects were observed by MCE. The RV wall thickness decreased, and the RV dimension increased, after RCA occlusion in each dog. The correlation of occlusion to baseline MBF ratios in the RV wall was closer to the ratio of b (r = 0.897, p = 0.0004) than A (r = 0.767, p = 0.0097) and was the closest to the ratio of Axb (r = 0.935, p < 0.0001). CONCLUSIONS: The RCA occlusion is manifested by RV wall thinning and dilation as well as by perfusion abnormalities consisting of patchy opacification defects by MCE. Myocardial contrast echocardiography-derived refilling parameters can be applied to assess RV perfusion abnormalities produced by RCA occlusion. PMID- 12767672 TI - Potent antifibrillatory effects of intrapericardial nitroglycerin in the ischemic porcine heart. AB - OBJECTIVES: We investigated the antiarrhythmic effects of intrapericardial nitroglycerin (NTG) during acute myocardial ischemia in the porcine heart. BACKGROUND: Nitroglycerin is a nitric oxide donor that exerts potent effects on the cardiovascular system. Intrapericardial administration allows investigation of pharmacologic actions on cardiac tissue in an in vivo system while minimizing the confounding influences of systemic effects. METHODS: In 29 closed-chest pigs, myocardial ischemia was induced by intraluminal balloon occlusion of the left anterior descending coronary artery. Arrhythmia incidence was monitored during 5 min balloon inflations performed without drug and at 15, 45, 75, and 105 min after NTG (4,000 microg bolus) administered by percutaneous transatrial access into the pericardial space. Electrocardiograms were monitored for ischemia induced T-wave alternans (TWA), a marker of electrical instability. The antiadrenergic potential of NTG was investigated by examining the drug's suppression of dobutamine-induced increase in myocardial contractility. RESULTS: Control coronary artery occlusion provoked ventricular fibrillation (VF) in all animals. Intrapericardial NTG suppressed VF at 45 min in all six pigs (p < 0.05) and reduced TWA across a parallel time course (from 459.1 +/- 144.4 microV before drug to 42.22 +/- 13.96 microV at 45 min, p = 0.047). The antifibrillatory effect occurred as early as 15 min and persisted for up to 75 min. Augmentation of maximum of the first time derivative of left ventricular pressure by dobutamine was blunted by intrapericardial NTG (from 3,999 +/- 196 mm Hg/s before NTG to 3,543 +/- 220 mm Hg/s at 15 min, p = 0.012). CONCLUSIONS: Intrapericardial NTG exerts a robust antifibrillatory action. Potential mechanisms include reduction in electrical instability and blunting of adrenergic effects. PMID- 12767673 TI - Will the nation need more cardiologists in the future than are being trained now? AB - This article supports the view that in the future our nation will need more general cardiologists who perform no high-tech procedures than we are currently training. It also addresses the reason graduates of American medical schools, especially women who make up 40% to 50% of the graduating class, are not choosing cardiology fellowship programs as they were previously. A remedy for this serious problem is offered. PMID- 12767675 TI - Hypertension and the prothrombotic state. PMID- 12767674 TI - Magnetic resonance imaging of targeted catheter-based implantation of myogenic precursor cells into infarcted left ventricular myocardium. AB - OBJECTIVES: This study was designed to test the hypothesis that myocardial implantation of myogenic precursor cells (MPC) loaded with iron oxide can be reliably detected in vivo by cardiac magnetic resonance imaging (MRI). BACKGROUND: In vivo imaging of targeted catheter-based implantation of MPC into infarcted left ventricular (LV) myocardium is unavailable. METHODS: The study was conducted in seven farm pigs (four with anterior myocardial infarction), in which autologous MPC were injected through a percutaneous catheter allowing for LV electromechanical mapping and guided micro-injections into normal and infarcted myocardium. Cardiac MRI was used to detect implanted MPC previously loaded with iron oxide nanoparticles. RESULTS: Magnetic resonance imaging data were compared with LV electromechanical mapping and cross-registered pathology. All 9 injections into normal and 12 injections into locally damaged myocardium were detected on T2-weighted spin echo and inversion-recovery true-fisp MRI (low signal areas) with good anatomical concordance with sites of implantation on electromechanical maps. All sites of injection were confirmed on pathology that showed in all infarct animals iron-loaded MPC at the center and periphery of the infarct as expected from MRI. CONCLUSIONS: Targeted catheter-based implantation of iron-loaded MPC into locally infarcted LV myocardium is accurate and can be reliably demonstrated in vivo by cardiac MRI. The ability to identify noninvasively intramyocardial cell implantation may be determinant for future experimental studies designed to analyze subsequent effects of such therapy on detailed segmental LV function. PMID- 12767677 TI - Questions remain regarding patients with aortic stenosis and severe pulmonary hypertension. PMID- 12767679 TI - Coronary atherosclerosis and body iron stores. PMID- 12767681 TI - Cost-effectiveness of a heart failure management program from the societal perspective? PMID- 12767683 TI - Use of spironolactone in heart failure patients receiving angiotensin-converting enzyme inhibitors and beta-blockers. PMID- 12767684 TI - Bisphenol A induces reactive oxygen species generation in the liver of male rats. AB - Bisphenol A, an environmental contaminant, widely used as a monomer in polycarbonate plastics, has been shown to cause abnormalities in liver of rats and mice. The nature and mechanism of action of bisphenol A on liver is not clear. The aim of the present study was to investigate if bisphenol A induces oxidative stress in the liver of rats and if co-administration of vitamin C, an antioxidant, can prevent oxidative stress. Bisphenol A (0.2, 2.0 and 20 micro g/kg body weight per day) and bisphenol A+vitamin C (0.2, 2.0, 20 micro g+40 mg/kg body weight per day) was orally administered to rats for 30 days. After 24 h of the last treatment, rats were killed using overdose of anesthetic ether. Body weights of the animals and the weights of liver showed no significant changes. The activities of antioxidant enzymes, superoxide dismutase, catalase, glutathione reductase and glutathione peroxidase were decreased in mitochondrial and microsome-rich fractions of liver. The levels of hydrogen peroxide and lipid peroxidation increased in the treated rats when compared with the corresponding group of control animals. Activity of alanine transaminase, a marker enzyme of hepatic injury remained unchanged in the treated rats as compared with the corresponding control rats. Co-administration of bisphenol A and vitamin C showed no changes in the activities of superoxide dismutase, catalase, glutathione reductase and glutathione peroxidase and in the levels of hydrogen peroxide and lipid peroxidation as compared with the corresponding control groups. The results indicated that bisphenol A induces oxidative stress in the liver of rats by decreasing the antioxidant enzymes. Co-administration of vitamin C reversed the effects of bisphenol A-induced oxidative stress in the liver of rats. PMID- 12767685 TI - Neurobehavioral evaluations of mixtures of trichloroethylene, heptachlor, and di(2-ethylhexyl)phthalate in a full-factorial design. AB - One approach to the toxicological evaluation of chemical mixtures is to construct full dose-response curves for each compound in the presence of a range of doses of each of the other compounds, i.e., a factorial design. This study was undertaken as part of an interdisciplinary project to evaluate a mixture of three environmental pollutants. A full-factorial design was undertaken to determine the neurobehavioral consequences of short-term repeated exposure to five dose levels each of three chemicals, in order to characterize potential two- and three-way interactions. Adult female F344 rats received (p.o.) for 10 days either one of five doses of trichloroethylene, di(2-ethylhexyl)phthalate, or heptachlor, or else one of all possible chemical combinations. Neurobehavioral evaluations were conducted using motor activity and an abbreviated functional observational battery. Response-surface analysis was applied to each of the endpoints. Hypotheses were tested based on the estimated model parameters; of primary interest was the overall test for interaction among the three chemicals. In addition, an abbreviated design was created by fitting only a subset of the data to the model. In general, significant overall interactions that deviated from response additivity were detected for most endpoints (11 of 14). All of the interactions on the neurobehavioral endpoints showed either antagonism, or else an interaction that could not be fully characterized. Often the results of the abbreviated dataset analysis were not the same as for the full-factorial design. This study was extremely intensive, in terms of the number of rats and time required for conduct of the study as well as the data analysis. These results underscore the need for more economical approaches to evaluate the toxic effects of mixtures of chemicals. PMID- 12767686 TI - Enhancement of carcinogen-induced malignant cell transformation by prostaglandin F(2 alpha). AB - The enhancement of carcinogen-induced malignant transformation of C3H/M2 mouse fibroblasts by the tumor promoters 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and 12-O-tetradecanoylphorbol-13-acetate (TPA) is associated with the induction of cyclooxygenase expression and the stimulation of prostaglandin (PG) formation. Therefore, the potential of PGs, i.e., PGF(2alpha) and PGE(2), for tumor promotion was studied in the two-step C3H/M2 cell transformation assay, a model of the multi-step process of carcinogenesis. The transformation of fibroblasts was clearly enhanced by the addition of PGF(2alpha) in the promotion phase after pretreatment with a subthreshold dose of a carcinogen (3-methylcholanthrene or N methyl-N'-nitro-N-nitrosoguanidine). No enhancement of cell transformation was observed in cells without carcinogen-pretreatment, i.e., PGF(2alpha) had no tumor initiating potential. The promotional effect was dose-dependent with a maximum at 16 nM PGF(2alpha). PGE(2) had no significant effect in this assay. Furthermore, PGF(2alpha) (but not PGE(2)) clearly reduced the inhibition of TPA-induced promotion by NS-398, an isozyme-specific inhibitor of cyclooxygenase-2. The inhibition of TPA- or TCDD-induced promotion by the non-specific cyclooxygenase inhibitor indomethacin was not affected by co-treatment with PGF(2alpha) and PGE(2). Our data suggest that PGF(2alpha) acts as an endogenous promoter of cell transformation implying that it may also be critically involved in tumor promoter induced signalling transfer cascades ultimately triggering the process of carcinogenesis. PMID- 12767687 TI - Cytotoxic mechanisms of hydrosulfide anion and cyanide anion in primary rat hepatocyte cultures. AB - Hydrogen sulfide and hydrogen cyanide are known to compromise mitochondrial respiration through inhibition of cytochrome c oxidase and this is generally considered to be their primary mechanism of toxicity. Experimental studies and the efficiency of current treatment protocols suggest that H(2)S may exert adverse physiological effects through additional mechanisms. To evaluate the role of alternative mechanisms in H(2)S toxicity, the relative contributions of electron transport inhibition, uncoupling of mitochondrial respiration, and opening of the mitochondrial permeability transition pore (MPTP) to hydrosulfide and cyanide anion cytotoxicity in primary hepatocyte cultures were examined. Supplementation of hepatocytes with the glycolytic substrate, fructose, rescued hepatocytes from cyanide anion induced toxicity, whereas fructose supplementation increased hydrosulfide anion toxicity suggesting that hydrosulfide anion may compromise glycolysis in hepatocytes. Although inhibitors of the MPTP opening were protective for hydrosulfide anion, they had no effect on cyanide anion toxicity, consistent with an involvement of the permeability transition pore in hydrosulfide anion toxicity but not cyanide anion toxicity. Exposure of isolated rat liver mitochondria to hydrosulfide did not result in large amplitude swelling suggesting that if H(2)S induces the permeability transition it does so indirectly through a mechanism requiring other cellular components. Hydrosulfide anion did not appear to be an uncoupler of mitochondrial respiration in hepatocytes based upon the inability of oligomycin and fructose to protect hepatocytes from hydrosulfide anion toxicity. These findings support mechanisms additional to inhibition of cytochrome c oxidase in hydrogen sulfide toxicity. Further investigations are required to assess the role of the permeability transition in H(2)S toxicity, determine whether similar affects occur in other cell types or in vivo and evaluate whether this may provide a basis for the design of more effective therapeutic measures for hydrogen sulfide intoxication. PMID- 12767688 TI - Metabolic interactions between acetylsalicylic acid and benzene. AB - The aim of the study was to evaluate cytochrome P-450 dependent hepatic monooxygenases system and urinary excretions of phenol and muconic acid in animals subjected to acetylsalicylic acid (ASA) orally and benzene by inhalations. ASA increased urinary excretion of muconic acid although it did not affect the urinary level of phenol. Benzene decreased concentrations of P-450 and b(5) cytochromes and the activities of NADPH-cytochrome P-450 and NADH-cytochrome b(5) reductases. In rats exposed to ASA and benzene simultaneously the concentration of both cytochromes and the activity of the cytochrome dependent reductases was higher than in the rats exposed only to benzene and sometimes exceeded the control group values. PMID- 12767689 TI - Influence of recipient gender on cytochrome P450 isoforms expression in intrasplenic fetal liver tissue transplants in rats. AB - Rat livers display a sex-specific cytochrome P450 (P450) isoforms expression pattern which is regulated by a differential profile of growth hormone (GH) secretion. The aim of the present study was to elucidate whether liver cell transplants at an ectopic site are also subject to this influence. Fetal liver tissue suspensions of mixed gender were transplanted into the spleen of adult male or female syngenic recipients. Four months after grafting transplant recipients and age-matched controls were treated with beta-naphthoflavone (BNF), phenobarbital (PB), dexamethasone (DEX) or the solvents and sacrificed 24 or 48 h thereafter. Livers and intrasplenic transplants were evaluated for the expression of the P450 subtypes 1A1, 2B1, 2E1, 3A2 and 4A1 by means of immunohistochemistry. The livers of both male and female rats displayed nearly no P450 1A1, but a distinct P450 2B1, 2E1, 3A2 and 4A1 expression. Whereas no sex differences were seen in the P450 1A1 expression, the immunostaining for P450 2B1, 3A2 and 4A1 was stronger in males and that for P450 2E1 in females. Similarly, in the intrasplenic liver cell transplants almost no P450 1A1, but a noticeable P450 2B1, 2E1, 3A2 and 4A1 expression was observed. Like in the respective livers, the immunostaining for P450 2B1, 3A2 and 4A1 was stronger in the transplants hosted by male than by female rats, whereas the opposite was the case for the P450 2E1 expression. Both in livers and transplants with some sex-specific differences P450 1A1 and 2E1 expression was induced by BNF, that of P450 2B1 by BNF and PB, and that of P450 3A2 by PB and DEX. These results indicate that the P450 system of ectopically transplanted liver cells is influenced by the gender of the recipient organism like that of the orthotopic livers. PMID- 12767690 TI - Primary DNA damage in chrome-plating workers. AB - In order to evaluate the primary DNA damage due to occupational exposure to chromium (VI), DNA strand-breaks and apoptosis in peripheral lymphocytes were measured in a group of 19 chrome-plating workers. DNA strand-breaks was assessed by alkaline (pH>13) single-cell microgel electrophoresis ('comet') assay, while apoptosis was measured by flow-cytometry after propidium iodide staining of the cells. Concentrations of chromium in urine, erythrocytes and lymphocytes were investigated as biological indicators of exposure. A group of 18 hospital workers (control group I) and another 20 university personnel (control group II) without exposure to chromium were also studied as controls. The results of the study show that chrome-plating workers have higher levels of chromium in urine, erythrocytes and lymphocytes than unexposed workers. Comet tail moment values, assumed as index of DNA damage, are increased in chromium-exposed workers and results are significantly correlated to chromium lymphocyte concentrations. No difference emerged in the percentage of apoptotic nuclei in exposed and unexposed workers. The study confirms that measurements of chromium in erythrocytes and lymphocytes may provide useful information about recent and past exposure to hexavalent chromium at the workplace. The increase in DNA strand-breaks measured by comet assay suggests this test is valid for the biological monitoring of workers exposed to genotoxic compounds such as chromium (VI). PMID- 12767691 TI - Comparative efficacy of diazepam and avizafone against sarin-induced neuropathology and respiratory failure in guinea pigs: influence of atropine dose. AB - This investigation compared the efficacy of diazepam and the water-soluble prodiazepam-avizafone-in sarin poisoning therapy. Guinea pigs, pretreated with pyridostigmine 0.1 mg/kg, were intoxicated with 4LD(50) of sarin (s.c. route) and 1 min after intoxication treated by intramuscular injection of atropine (3 or 33.8 mg/kg), pralidoxime (32 mg/kg) and either diazepam (2 mg/kg) or avizafone (3.5 mg/kg). EEG and pneumo-physiological parameters were simultaneously recorded. When atropine was administered at a dose of 3 mg/kg, seizures were observed in 87.5% of the cases; if an anticonvulsant was added (diazepam (2 mg/kg) or avizafone (3.5 mg/kg)), seizure was prevented but respiratory disorders were observed. At 33.8 mg/kg, atropine markedly increased the seizure threshold and prevented early respiratory distress induced by sarin. When diazepam was administered together with atropine, seizures were not observed but 62.5% of the animals displayed respiratory difficulties. These symptoms were not observed when using avizafone. The pharmacokinetic data showed marked variation of the plasma levels of atropine and diazepam in different antidote combination groups, where groups receiving diazepam exhibited the lowest concentration of atropine in plasma. Taken together, the results indicate that avizafone is suitable in therapy against sarin when an anticonvulsant is judged necessary. PMID- 12767692 TI - Inhibition of glycyrrhizic acid on aflatoxin B1-induced cytotoxicity in hepatoma cells. AB - Aflatoxin B(1) (AFB(1)) causes oxidative stress and ROS formation via metabolic activation of AFB(1). Glycyrrhizic acid (GA) has been reported to have antioxidative properties. The present study was to investigate the effect of GA, a major component of licorice on AFB(1)-induced cytotoxicity in human hepatoma cell line (HepG2). GA displayed protective effects against AFB(1) treatment. Both CYP1A1, and glutathione S-transferase (GST) activities were increased in cells after treatment with the GA, while CYP1A2 did not seem to be affected by GA. For cells without GA pre-treatment, cell injury was implicated as indicated by the decrease in cell viability. The time-course study of GA showed pretreatment of cells with GA for 24 h was effective. The treatment of cells with GA and AFB(1) enhanced the detoxifying enzyme activity. The pre-treatment of cells with GA provides protective effects in terms of the enzyme activity and increase in cell viability. The results suggest that GA protects against aflatoxin-induced oxidative stress. This may contribute to its anticarcinogenic capability. The protective effect is likely due to its capacity to inhibit the metabolic activation of hepato-toxin, a critical factor in the pathogenesis of chemical induced carcinogenicity. PMID- 12767693 TI - Characterization of the in vitro kinetic interaction of chlorpyrifos-oxon with rat salivary cholinesterase: a potential biomonitoring matrix. AB - The primary mechanism of action for organophosphorus (OP) insecticides such as chlorpyrifos (CPF) involves the inhibition of acetylcholinesterase (AChE) by their active oxon metabolites resulting in a wide range of neurotoxic effects. These oxons also inhibit other cholinesterases (ChE) such as butyrylcholinesterase (BuChE), which represents a detoxification mechanism and a potential biomarker for OP insecticide exposure/response. Salivary biomonitoring has recently been explored as a practical method for examination of chemical exposure, however, there are few studies exploring the use of saliva for OP insecticides. To evaluate the use of salivary ChE as a biological monitor for OP insecticide exposure, a modified Ellman assay in conjunction with a pharmacodynamic model was used to characterize salivary ChE in adult male Sprague Dawley rats. Comparison of rat saliva, brain, and plasma ChE activity in the presence of selective inhibitors of AChE and BuChE (BW284C51 and iso-OMPA, respectively) with different ChE substrates indicated that rat salivary ChE activity is primarily associated with BuChE (>95%). Further characterization of rat salivary BuChE kinetics yielded an average total BuChE active site concentration of 1.20+/-0.13 fmol ml(-1) saliva, an average reactivation rate constant (Kr) of 0.070+/-0.008 h(-1), and an inhibitory rate constant (Ki) of approximately 9 nM(-1) h(-1). The pharmacodynamic model successfully described the in vitro BuChE activity profile as well as the kinetic parameters. These results support the potential utility of saliva as a biomonitoring matrix for evaluating occupational and environmental exposure to CPF and other OP insecticides. PMID- 12767694 TI - Drinking water exposure to cadmium, an environmental contaminant, results in the exacerbation of autoimmune disease in the murine model. AB - Cadmium is a pervasive environmental contaminant. The primary route of exposure to the general population occurs via contaminated drinking water or food supplies. Our hypothesis was that cadmium could be a trigger for inducing autoimmune disease (AD) in genetically predisposed populations. Therefore, New Zealand Black/White F1 (NZBW) mice were exposed to cadmium via drinking water. Mice were exposed to: 0, 3, 30, 3000 or 10000 parts per billion (ppb) of cadmium in tap water for 2, 4, 28, or 31 weeks. After 4 weeks of exposure, in the group of mice exposed to 10000 ppb cadmium, there was an increased incidence of antinuclear antibodies (ANA). There was also deposition of immune complexes in all groups after 4 weeks of exposure. After 31 weeks, there were increases in IgG2a in mice exposed to low doses of cadmium. In an attempt to establish the progression from an autoimmune reaction to the development of AD, the biological marker for AD, proteinuria, was assessed. Onset of proteinuria was exacerbated by 11 weeks in mice exposed to cadmium. This data suggests that short-term exposure may result in a type of autoimmune reaction since the mice are beginning to produce ANA after only 4 weeks of exposure and there is immune-complex deposition in the kidney. Long-term exposure to cadmium appears to result in the exacerbation of AD as indicated by the development of proteinuria and continued presence of immune complexes in the kidney. The mechanism may involve the increased production of IgG2a, which is capable of forming immune complexes and causing autoimmune glomerulonephritis. PMID- 12767695 TI - Effect of lead exposure on the immune response of some occupationally exposed individuals. AB - Lead is a ubiquitous pollutant in the industrial environment, which poses serious threats to human health. In the past 20 years increasing attention has been paid to the effects of lead exposure on health. This toxic metal alters the immune response of animals as well as humans. To study the immunological effects of occupational exposure to lead, we examined lymphocyte proliferation, natural killer (NK) cell cytotoxicity and interferon-gamma production with peripheral blood mononuclear cells (PBMCs) of individuals occupationally exposed to lead. We selected three different groups of individuals exposed to lead: three-wheeler drivers (30), battery workers (34) and silver jewelery makers (20); and unexposed healthy volunteers (30) as control for comparison. Our results indicate that though lymphocyte proliferation to phytohaemagglutinin (PHA) is inhibited in lead exposed individuals as compared with unexposed volunteers, there is no correlation between inhibition of lymphocyte proliferation and blood lead level. NK cell cytotoxicity remains unaffected in individuals exposed to lead as compared with controls. On the other hand, we observed that interferon-gamma (IFN gamma) was significantly elevated in T cell mitogen, PHA, stimulated PBMCs culture supernatant of lead exposed individuals. We found significant positive correlation between blood lead levels and IFN-gamma produced in culture supernatant on stimulation with PHA. In brief, this study demonstrates that lead can affect the immune response of the occupationally exposed individuals such as three-wheeler drivers, battery reconditioning workers and silver jewelery makers. PMID- 12767696 TI - Comparative cytotoxicity of dimethylamide-crotonin in the promyelocytic leukemia cell line (HL60) and human peripheral blood mononuclear cells. AB - Dehydrocrotonin (DHC) is a diterpene lactone obtained from Croton cajucara (Sacaca). Dimethylamide-crotonin (DCR), a DHC derivative, has a similar inhibitory effect on leukemic HL60 cells than its parent compound evaluated by different endpoints of cytotoxicity. No cytotoxicity or morphological alterations associated with apoptosis were detected in human peripheral blood mononuclear cells (PBMC) after treatment with up to 400 micro M DCR in presence of phytohemaglutinin (5 micro g/ml). Based on morphological changes and the pattern of DNA fragmentation, DHC and DCR were found to induce apoptosis and terminal differentiation (assessed by nitro blue tetrazolium reduction) in HL60 cells, but these compounds did not show any toxic effect in PBMC. Thus, DCR and DHC inhibit HL60 cell growth in vitro partly by inducing apoptosis and cell differentiation, but does not cause serious damage to immune cells according to our experimental conditions. PMID- 12767697 TI - Dose-threshold for thyroid tumor-promoting effects of xylazine in rats. AB - To clarify the threshold dose of thyroid tumor-promoting effects of xylazine hydrochloride (XZ), male F344 rats received pulverized basal diet containing 0, 250, 500, or 1000 ppm XZ for 26 weeks with or without initiation of 2400 mg/kg N bis(2-hydroxypropyl)nitrosamine (DHPN). Thyroid weights significantly increased in the groups with or without DHPN initiation that were given 500 ppm XZ or more. The serum thyroxine (T4) and triiodothyronine (T3) levels decreased significantly in the XZ 250 and XZ 1000 ppm groups, respectively, although there were no remarkable changes in the serum thyroid-stimulating hormone (TSH) levels. Histopathologically, follicular cell hyperplasias and adenomas were induced in the DHPN-alone and DHPN+XZ groups, and the incidences and multiplicities of these lesions in the DHPN groups treated with 500 ppm XZ or more were significantly higher than those in the DHPN alone group. These results suggest that the threshold dose of rat thyroid tumor-promoting effects of XZ is between 250 and 500 ppm under the present experimental condition. PMID- 12767698 TI - Comparative toxicity evaluation of cyanobacterial cyclic peptide toxin microcystin variants (LR, RR, YR) in mice. AB - The cyclic peptide toxins microcystins and nodularins are the most common and abundant cyanotoxins present in diverse water systems. They have been the cause of human and animal health hazards and even death. Over 60 microcystin variants have been reported so far. We report here the results of our study on comparative toxicity evaluation of three most predominant microcystins, MC-LR, MC-RR and MC YR in mice. The mice were administered one LD(50) dose of MC-LR, RR and YR (43, 235.4 and 110.6 micro g/kg body weight, respectively), and biochemical and histological variables were determined at 30 min post-treatment and mean time to death (MTD). Significant increase in liver body weight index was induced by all three variants. There was marginal increase in serum levels of hepatic enzymes viz. AST, ALT and gamma-GT at 30 min post-treatment but 3-4 fold increase was observed at MTD. In contrast, enhanced LDH leakage, DNA fragmentation and depletion of hepatic glutathione was observed at 30 min post treatment in all three variants. There was no change in levels of serum protein, albumin and albumin/globulin ratio. Liver histology showed time dependent severe pathological lesions like congestion, haemorrhage, portal mononuclear cell infiltration and obliteration of chromatin material. Lung lesions were predominantly in bronchi and parenchyma. Though qualitatively lesions were identical in all three microcystin variants, degree of liver and lung lesions varied quantitatively with the toxin. The breathing pattern and respiratory frequency of the mice after i.p. administration of the toxin showed uniform pattern for 90 min followed by abrupt change in the respiratory pattern and instantaneous death. Based on biochemical and histological studies, MC-LR was found to be the most potent toxin followed by MC-YR and MC-RR. PMID- 12767699 TI - Safety assessment for genetically modified sweet pepper and tomato. AB - The coat protein (CP) gene of cucumber mosaic virus (CMV) was cloned from a Chinese CMV isolate, the CaMV promoter and NOS terminator added and the gene construct was transformed into both sweet pepper and tomato plants to confer resistance to CMV. Safety assessments of these genetically modified (GM) plants were conducted. It was found that these two GM products showed no genotoxicity either in vitro or in vivo by the micronucleus test, sperm aberration test and Ames test. Animal feeding studies showed no significant differences in growth, body weight gain, food consumption, hematology, blood biochemical indices, organ weights and histopathology between rats or mice of either sex fed with either GM sweet pepper or tomato diets compared with those with non-GM diets. These results demonstrate that the CMV-resistant sweet pepper and tomato are comparable to the non-GM counterparts in terms of food safety. PMID- 12767700 TI - The effect of endotoxin on the production of IgE, IgG1 and IgG2a antibodies against the cat allergen Fel d 1 in mice. AB - BACKGROUND: Endotoxin/LPS is ubiquitous in our environment. The question whether lipopolysaccharide (LPS) is beneficial or disease-promoting in relation to asthma and allergy has been raised in several recent studies. Some have reported a positive correlation between the level of LPS in house dust and the symptoms of asthmatic children. Others have found that exposure to LPS appears to protect against the development of atopic disease in children. OBJECTIVES: We performed a study in mice to examine the antibody response after subcutaneous immunization with LPS and the cat allergen Fel d 1. We asked whether LPS would increase the response and direct the antibody production towards an allergic (IgE), or non allergic (IgG2a) antibody profile. In rodents both IgE and IgG1 are antibodies produced under Th2-dependence and IgG2a antibodies under Th1-dependence. Also, when LPS and Fel d 1 are introduced to the immune system, we asked whether the timing of the two agents relative to each other is crucial. METHODS: The mice were injected subcutaneously with LPS and/or Fel d 1 four times in various orders. IgE, IgG1 and IgG2a antibodies specific to Fel d 1 were measured in serum using ELISA. RESULTS: A strong antibody response, both for IgE, IgG1 and IgG2a, was observed only when Fel d 1 and LPS were injected simultaneously, and in particular after repeated injections. CONCLUSION: A strong specific antibody response was observed, both for IgE, IgG1 and IgG2a, only when LPS was introduced to the immune system together with the cat allergen Fel d 1. No such adjuvant effect was observed when LPS was introduced alone prior to or subsequent to the allergen. The resulting antibody response was not polarized in terms of Th1- or Th2-dependence. PMID- 12767701 TI - Inhibition of LPS-induced splenocyte proliferation by ortho-substituted polychlorinated biphenyl congeners. AB - Polychlorinated biphenyls (PCBs) are persistent environmental contaminants, and their ubiquitous nature has prompted studies of their potential health hazards. As a result of their lipophilic nature, PCBs accumulate in breast milk and subsequently affect the health of offspring of exposed individuals. Biological effects of PCBs in animals have mostly been attributed to coplanar congeners, although effects of ortho congeners also have been demonstrated. To investigate the relationship of immunotoxicity and chlorine substitution pattern, the effects of PCB congeners and mixtures of ortho and non-ortho-substituted constituents of Aroclor 1242 on splenocytes from C57B1/6 mice were examined. The immunotoxic endpoints investigated included splenocyte viability, lipopolysaccharide (LPS) induced splenocyte proliferation, and LPS-induced antibody secretion. Congeners with multiple ortho chlorines preferentially inhibited splenocyte proliferation as compared with non- or mono-ortho-substituted congeners. However, mixtures of non- and mono-ortho-substituted congeners and multi-ortho-substituted congeners inhibited LPS-induced splenocyte proliferation and antibody secretion at similar concentrations. Exposure of splenocytes to these mixtures did not activate the aryl hydrocarbon receptor (AhR) signal transduction pathway. These results suggest individual multi-ortho-substituted congeners preferentially inhibit LPS induced splenocyte proliferation, while congeners not exhibiting an effect individually may have additive effects in a mixture to produce an immunotoxic response through an AhR-independent pathway. PMID- 12767703 TI - Controlled drug delivery from swellable hydroxypropylmethylcellulose matrices: model-based analysis of observed radial front movements. AB - This work is related to the on-going development of mathematical models describing transient drug delivery from hydroxypropylmethylcellulose (HPMC) matrices. Recently, experimental data providing a detailed mapping of radial swelling, diffusion, and erosion front movements in a high-viscosity HPMC matrix were published [J. Controlled Release 70 (2001) 383]. Using these and other data for verification of simulations, a detailed mathematical model, taking into account water-induced swelling, drug dissolution, and external and internal mass transport resistances of dissolved drug, has been developed. In contrast to earlier models, explicit equations for the rate of movement of the swelling, diffusion and erosion fronts, with the relevant physical properties of drug and HPMC matrix contained in the equations, were derived. Simulations have been compared to transient experimental data for three drugs of very different water solubility and a good agreement was found, taking into account the uncertainty of key input parameters. Furthermore, the model predicts the presence of the drug particle translocation phenomenon observed experimentally. However, continued swelling of the matrix, subsequent to the disappearance of the swelling front, could not be described by the present model. The front-tracking approach illustrated is of relevance in the development of detailed and accurate models of drug delivery from swellable cylindrical matrices involving both axial and radial diffusion. PMID- 12767704 TI - Modeling and analysis of dispersed-drug release into a finite medium from sphere ensembles with a boundary layer. AB - Mathematical models were developed and analytical solutions were derived for describing kinetics of dispersed-drug release into a finite external medium from multi-particulate systems, such as ensembles of matrix spheres and microcapsules with a diffusion boundary layer. The solutions can be used to compute profiles of the moving boundary of a dispersed drug and the amount of drug released for multiparticulate ensembles with various ratios of initial drug loading (C(0)) to drug solubility (C(s)) in a finite to infinite medium. They are also applicable to a single sphere without a boundary layer in a perfect sink. The determinants of release kinetics, such as the liquid volume, the initial drug loading, the boundary layer thickness, and the number of spheres in a population, were analyzed using the derived solutions. The effect of coating thickness and material on the release profiles of microcapsules was studied as well. Criteria were established for finding the conditions when drug release would stall due to saturation of the medium, which can be used to determine suitable liquid volume and time for refreshing the medium. PMID- 12767706 TI - Combined chemical and electrical enhancement modulates stratum corneum structure. AB - In a previous in vitro study it has been shown that pretreatment with a water based surfactant formulation results in a two-fold increase in transdermal iontophoretic transport of R-apomorphine compared to iontophoresis only. The aim of the study presented in this paper was to unravel the mechanisms involved in the increased iontophoretic delivery. Freeze fracture electron microscopy and cryo-scanning electron microscopy were used to visualise the ultrastucture of human stratum corneum after (i) application of the surfactant formulation, (ii) iontophoresis and (iii) application of the surfactant formulation followed by iontophoresis. Non-occlusive application of the surfactant formulation did not exert any detectable changes in the ultrastructure of the stratum corneum, except for swelling of the outermost corneocyte layers. Application of a current density of 0.5 mA/cm(2) for 9 h induced a swelling of the corneocytes and the formation of water pools that were occasionally present in the intercellular regions. Application of the surfactant formulation followed by iontophoresis resulted in a further swelling of the corneocytes and a frequent presence of water pools in the intercellular regions throughout the whole stratum corneum. The observed changes in the ultrastructure of the stratum corneum can explain the increased R apomorphine transport during iontophoresis. PMID- 12767705 TI - Relationships between the hydrophilic-lipophilic balance values of pharmaceutical excipients and their multidrug resistance modulating effect in Caco-2 cells and rat intestines. AB - The effects of a series of pharmaceutical excipients, including Span 80, Brij 30, Tween 20, Tween 80, Myrj 52, and sodium lauryl sulfate (with increasing hydrophilic-lipophilic balance (HLB) values) on the intracellular accumulation, transport kinetics, and intestinal absorption of epirubicin were investigated in both the human colon adenocarcinoma (Caco-2) cell line and the everted gut sacs of rat jejunum and ileum. The possible use of these excipients as multidrug resistance (MDR) reversing agents also was examined. Epirubicin uptake experiments using a flow cytometer showed that these selected excipients markedly enhanced the intracellular accumulation of epirubicin in Caco-2 cells in a dose dependent manner. The optimal effect on the epirubicin uptake was characteristic of excipients with intermediate HLB values ranging from 10 to 17. Moreover, the optimal net efficacy was observed for excipients with polyoxyethylene chains and intermediate chain length of fatty acid and fatty alcohol (monolaurate for Tween 20, monooleate for Tween 80, monostearate for Myrj 52, and lauryl alcohol for Brij 30). These excipients significantly increased apical to basolateral absorption and substantially reduced basolateral to apical efflux of epirubicin across Caco-2 monolayers. Furthermore, the addition of Tween 20, Tween 80, Myrj 52, and Brij 30 markedly enhanced mucosal to serosal absorption of epirubicin in the rat jejunum and ileum. This study suggests that inhibition of intestinal P glycoprotein (P-gp), multidrug resistance associated protein family (MRPs), or other transporter proteins by pharmaceutical excipients may improve oral absorption of drugs in MDR spectrum. The optimal HLB values of surfactant systems with suitable hydrocarbon chains and polar groups are an important factor in designing promising epirubicin formulations for reversing MDR. In conclusion, therapeutic efficacy of epirubicin may be enhanced by the use of such low toxicity excipients as absorption enhancers and MDR modulators in formulations. This provides a potential strategy for improving bioavailability in the optimization of formulations for drugs performing intestinal absorption and secretion. PMID- 12767707 TI - Investigation of the release behavior of a covered-rod-type formulation using silicone. AB - The purpose of this study was to investigate the effects of the properties of a drug on its release behavior in a cylindrical sustained-release formulation having a two-component structure, with a silicone matrix containing drug powder as the inner layer component, and with its lateral side covered with an silicone outer layer (CR silicone formulation). In this study, the release profile of a drug from "the lateral side covered with silicone" and from "the cross-sections where the inner layer is exposed to the surface" was examined using a newly designed bi-directional elution cell. The relationships between the release profile and solubility of the drug and its permeability through silicone were also studied. Bovine serum albumin (BSA), antipyrine (ANP), indometacin (IDM) and ketoprofen (KP) were used as model drugs. Each CR silicone formulation containing drug powder consisting of a drug and sucrose (SUC) was investigated, and a satisfactory relationship was observed between drug release from the cross sections and drug solubility, and between drug release from the lateral side and permeability of the drug through a silicone membrane. For CR silicone formulations containing IDM, the addition of deoxycholate sodium (DOC) improved the solubility of IDM; however, release from the lateral side of the formulation remained unchanged, and IDM release from the cross-sections of the formulation increased. In this study it was found that, for controlled release of a drug from CR silicone formulations, control of drug solubility is effective. PMID- 12767708 TI - In vitro characteristics of liposomes and double liposomes prepared using a novel glass beads method. AB - A novel preparative method for liposomes and double liposomes (DL) using glass beads was superior to a glass-filter method developed previously. Lipid dissolved in chloroform was poured into a kjeldahl flask with glass beads (BZ-04, 0.350 0.500 mm phi; BZ-3, 2.794-3.962 mm phi; or BZ-6, 5.613-6.680 mm phi), and the organic solvent was evaporated. The lipid layer that formed on the glass beads was hydrated with 1.5 ml of the suspension of inner liposomes at a temperature above the phase transition temperature of the lipids employed, and was agitated vigorously. Erythrosine (ER) was used as a model drug. The size of liposomes prepared by the glass beads method depended on the size of the glass beads. The size of the liposomes became smaller as glass beads with a smaller size were used. A high encapsulation efficiency was observed when glass bead blends consisting of two different sizes were used. Large sizes (BZ-3/BZ-6) had a tendency to show high encapsulation efficiency and size also played an important role in the formation of liposomes. DL formation inhibited the release of ER and DL formative efficiency was markedly improved by means of the glass beads method. These findings suggested that the glass beads method developed in this study conferred a high drug loading and a high DL formation on liposomes compared with ordinary methods. PMID- 12767709 TI - Characterization of DNA-hyaluronan matrix for sustained gene transfer. AB - DNA-Hyaluronan (DNA-HA) matrix formulations intended for use as gene delivery systems have been developed and their potential for delivering DNA encoding a model therapeutic cytokine, platelet-derived growth factor (PDGF), has been evaluated. The results of enzyme-mediated release kinetics studies suggested that the rate of DNA release from the DNA-HA matrices could be modulated by changing the DNA loading or the degree of crosslinking. SEM imaging of the DNA-HA matrix showed that it was gradually eroded by enzymatic action. The results of gel electrophoresis suggested that there was some degree of interaction between DNA and native HA and that portions of the DNA released from the DNA-HA matrices were associated with crosslinked HA fragments. Only fractions of the DNA released from the DNA-HA matrices were free and the rest was entrapped by HA fragments, which could serve as a mechanism for DNA protection. The results from cell transfection studies using DNA samples collected during the course of release studies confirmed this hypothesis. The PDGF produced by transfection of the DNA released from DNA-HA matrices induced human dermal fibroblast cells to proliferate. PMID- 12767710 TI - A general approach to describe the antimicrobial agent release from highly swellable films intended for food packaging applications. AB - A mathematical model able to describe the release kinetics of antimicrobial agents from crosslinked polyvinylalcohol (PVOH) into water is presented. The model was developed by taking into account the diffusion of water molecules into the polymeric film, the counter-diffusion of the incorporated antimicrobial agent from the film into water, and the polymeric matrix swelling kinetic. To validate the model the water sorption kinetics as well as the release kinetics of three antimicrobial agents (i.e., lysozyme, nisin and sodium benzoate, all approved to be used in contact with food) were determined at ambient temperature (25 degrees C). The three investigated active agents were entrapped in four films of PVOH with a different degree of crosslink. The model was successfully used to fit all the above sets of data, corroborating the validity of the hypothesis made to derive it. PMID- 12767711 TI - Development of a new dissolution test method for an oral controlled release preparation, the PVA swelling controlled release system (SCRS). AB - In vitro dissolution tests of novel controlled release tablets, the poly(vinyl alcohol) (PVA) swelling controlled release system (SCRS), were performed by various methods under different conditions in the sinking state in water. The in vitro release profiles of various tests were almost the same and faster than the in vivo absorption profiles calculated from the plasma drug concentrations of humans. A novel dissolution test method was developed considering the gastrointestinal tract (GI) conditions. PVA particles were used as the filler in a flow-through cell. PVA particles swelled with water were put in the flow through cell and the tablet was buried in PVA. The test medium was dropped from the top of the cell, and the dissolution medium that dripped from the bottom of the cell was collected and assayed. The in vitro dissolution profile determined by this method was similar to the in vivo absorption profile against other in vitro methods in the sinking state in water. Furthermore, a good correlation between in vitro and in vivo for the two different release rate preparations was obtained using a flow pattern imitating the GI transition. PMID- 12767713 TI - Cell microencapsulation technology for biomedical purposes: novel insights and challenges. AB - The aim of cell microencapsulation technology is to treat multiple diseases in the absence of immunosuppression. Using this technique, cells are immobilized within carefully designed capsules that allow the long-term function of the graft. Although the potential impact of this field is likely to be wide-ranging, the past few years have seen several 'firsts' that have brought the whole technology much closer to a realistic clinical application. PMID- 12767712 TI - Complex dispersions of multilamellar vesicles: a promising new carrier for controlled release and protection of encapsulated molecules. AB - Multilamellar vesicles called Spherulites have recently been discovered and are being developed for encapsulation applications. In this study, we present new systems of Spherulites called complex dispersions. These are prepared by dispersing Spherulites within an oily medium, and then emulsifying this oily dispersion of Spherulites within an aqueous solvent. The ability of complex dispersions to reduce the release of encapsulated ions under variable osmotic dilutions was evaluated and compared with Spherulites directly dispersible in an aqueous medium, and with multiple emulsions. An advantage of complex dispersions over Spherulites is to present an additional oily barrier. Indeed, this barrier retarded the release of encapsulated ions. Complex dispersions also proved to be less sensitive to osmotic pressure than multiple emulsions. It appeared that the dilution of a complex dispersion formulated with no external aqueous phase containing a hydrophilic surfactant provided the slowest release of encapsulated ions. Furthermore, this formulation maintained a difference of pH between the internal and external aqueous phases for a few hours. In conclusion, these new systems of Spherulites known as complex dispersions show great potential for pharmaceutical applications such as controlled release and protection of encapsulated substances. PMID- 12767714 TI - Can a proteomics strategy be used to identify the anti-malarial activity of chloroquine? AB - Interest in the mechanism of action of chloroquine is intense partly because of the emergence of drug-resistant parasites. Chloroquine resistance has been genetically linked to mutations in a parasite protein (PfCRT) that might confer resistance by inhibiting chloroquine accumulation in infected erythrocytes. Now chloroquine-binding proteins in malaria-infected erythrocytes, surprisingly, have been identified as human aldehyde dehydrogenase 1 and quinone reductase 2, raising the interesting possibility that the target of the anti-malarial activity of chloroquine might be a host enzyme. PMID- 12767716 TI - Protein transduction domains: are they delivering? AB - Early studies with protein transduction domains (PTDs), such as those derived from Tat and Drosophila Antennapedia, showed rapid, receptor- and endosomal independent uptake of conjugated biological tools into all cell types. However, recent mechanistic studies suggest that these observations were artefacts of the positively charged nature of PTDs and that uptake is instead via electrostatic binding to the plasma membrane and subsequent endocytosis. Given these observations, we assess the future utility of PTDs for in vitro and in vivo cellular delivery. PMID- 12767717 TI - Protein transduction technology offers novel therapeutic approach for brain ischemia. AB - Transient or permanent reduction in cerebral blood flow following ischemia can lead to severe and irreversible tissue damage to the brain. Emerging biochemical evidence suggests a role for apoptosis in neuronal death following cerebral ischemia. Despite the abundance of studies on the subject, therapeutic interventions for ischemia-related cell injury have so far proved disappointing in clinical trials. Recently, four new, exciting studies reported the use of protein transduction technology to deliver anti-apoptotic molecules to protect neuronal cells following ischemic stroke in vivo. These studies offer new avenues for the treatment and prevention of cell death following brain injuries. PMID- 12767718 TI - Herbal medicine, cancer prevention and cyclooxygenase 2 inhibition. PMID- 12767719 TI - Investigator bias and false positive findings in medical research. PMID- 12767720 TI - Novel interactions between K+ channels and scorpion toxins. AB - K(+) channels are macromolecules embedded in biological membranes, where they play a key role in cellular excitability and signal transduction pathways. Knowledge of their structure should help improve our understanding of their function and lead to the design of therapeutic compounds. Most pharmacological and structural characteristics of these channels have been elucidated by using high-affinity channel blockers isolated from scorpion venoms. Recent data on the three-dimensional structures of K(+) channels and novel scorpion toxins suggest a variety of novel interacting modes of these channels and toxins, which should help increase our understanding of the K(+) channel structure-function relationship. PMID- 12767721 TI - Nicotinamide: necessary nutrient emerges as a novel cytoprotectant for the brain. AB - Although usually identified as an essential cellular nutrient for cellular growth and maintenance, nicotinamide is under development as a novel cytoprotectant for acute and chronic neurodegenerative disorders. Here, we outline support for the premise that nicotinamide both prevents and reverses neuronal and vascular cell injury. Nicotinamide fosters DNA integrity and maintains phosphatidylserine membrane asymmetry to prevent cellular inflammation, cellular phagocytosis and vascular thrombosis. The downstream cellular and molecular cascades are considered vital for the cytoprotection offered by nicotinamide. These pathways encompass the modulation of Akt, the forkhead transcription factor FKHRL1, mitochondrial membrane potential, caspase activities and cellular energy metabolism, but remain independent of intracellular pH and mitogen-activated protein kinases. As both a therapeutic agent and an investigational tool, nicotinamide offers new therapeutic strategies for degenerative disorders of the CNS. PMID- 12767722 TI - The Wnt pathway, cell-cycle activation and beta-amyloid: novel therapeutic strategies in Alzheimer's disease? AB - Beta-amyloid protein (betaAP) is thought to cause neuronal loss in Alzheimer's disease (AD). Applied to neurons in culture, betaAP induces neuronal death and hyperphosphorylation of tau protein, which forms neurofibrillary tangles (NFTs) in AD brains. Neurons also undergo rapid apoptotic death following reactivation of a mitotic cycle. However, the molecular events that determine the fate of neurons challenged with betaAP (apoptotic death, formation of NFTs and survival) are unclear. We discuss a scenario for the pathogenesis of AD. This links betaAP induced changes to the Wnt signaling pathway that promotes proliferation of progenitor cells and directs cells into a neuronal phenotype during brain development. We propose that betaAP-mediated facilitation of mitogenic Wnt signaling activates unscheduled mitosis in differentiated neurons. Furthermore, late downregulation of Wnt signaling by betaAP might lead to NFT formation. We propose that drugs that both inhibit the cell cycle and rescue Wnt activity could provide novel AD therapeutics. PMID- 12767723 TI - A central role of EGF receptor transactivation in angiotensin II -induced cardiac hypertrophy. AB - In addition to their physiological roles in the cardiovascular system (CVS), G protein-coupled receptor (GPCR) agonists such as noradrenaline, endothelin-1 and angiotensin II (Ang II) are known to be involved in the development of cardiac hypertrophy. Recent studies using targeted overexpression of the angiotensin AT(1) receptor in cardiomyocytes suggest that Ang II can directly promote the growth of cardiomyocytes via transactivation of the epidermal growth factor (EGF) receptor and subsequent activation of mitogen-activated protein kinases (MAPKs). This process is mediated by the production of heparin-binding EGF (HB-EGF) by metalloproteases. Blockade of the generation of HB-EGF by metalloprotease inhibitors, or abrogation of EGF receptor kinase activity by selective pharmacological inhibitors or antisense oligonucleotides, protects against Ang II mediated cardiac hypertrophy. These approaches offer a potential therapeutic strategy to prevent cardiac remodeling and hypertrophy, and possibly prevent progression to heart failure. PMID- 12767724 TI - Prevention of thrombosis and vascular inflammation: benefits and limitations of selective or combined COX-1, COX-2 and 5-LOX inhibitors. AB - Anti-thrombotic therapy with aspirin, which at low doses acts as a selective inhibitor of platelet cyclooxygenase 1 (COX-1) activity, is well established. However, a major limitation of aspirin treatment is its gastrointestinal toxicity, which is thought to be linked to the suppression of COX-1-mediated production of cytoprotective prostaglandins. Selective COX-2 inhibitors are effective anti-inflammatory agents with lower gastrointestinal toxicity than aspirin. These inhibitors might also downregulate vascular and leukocyte inflammatory components that play a major part in atherothrombotic disease. However, some selective COX-2 inhibitors appear to increase cardiovascular risk. Newly developed dual COX-5-lipoxygenase (5-LOX) inhibitors share the anti inflammatory effect and gastric safety of COX-2 inhibitors, but also inhibit COX 1-mediated platelet function and 5-LOX-mediated synthesis of gastrotoxic leukotrienes. Dual inhibitors might thus be beneficial in the treatment of atherosclerosis, where platelet-leukocyte interaction dominates the underlying inflammatory process. PMID- 12767725 TI - The importance of being varied in steroid receptor transactivation. AB - A major response of steroid receptors to steroid hormones is the induction of gene transcription. Two relevant, albeit less studied, properties of these receptors are the EC(50) values of the receptor-agonist complexes and the partial agonist activity of the receptor-antagonist complexes. Contrary to earlier expectations, neither the EC(50) value nor the partial agonist activity is constant for a given receptor-steroid complex. This variation is, however, beneficial to cells and organisms because it provides a mechanism both for differential control of gene expression by a single concentration of circulating hormone and for limiting side-effects during endocrine therapies. In this article, the factors and proposed mechanisms for the modulation of the EC(50) value and partial agonist activity of receptor-steroid complexes are discussed. PMID- 12767726 TI - A new vision for human security. PMID- 12767727 TI - Topiramate for the treatment of alcohol dependence: initiating abstinence. PMID- 12767728 TI - Should pregnant women avoid eating fish? Lessons from the Seychelles. PMID- 12767729 TI - Hyperhomocysteinaemia and vascular disease--a role for DNA hypomethylation? PMID- 12767730 TI - Tobacco and the Commonwealth: a call to action. PMID- 12767731 TI - Effect of genotype on phenotype and mortality in cystic fibrosis: a retrospective cohort study. AB - BACKGROUND: Over 1000 mutations of the cystic fibrosis transmembrane conductance regulator gene (CFTR) that cause cystic fibrosis have been identified. We examined the effect of CFTR genotype on mortality and disease phenotype. METHODS: Using the US Cystic Fibrosis Foundation National Registry, we did a retrospective cohort study to compare standardised mortality rates for the 11 most common genotypes heterozygous for DeltaF508 with those homozygous for DeltaF508. Of the 28455 patients enrolled in the registry at the time of our analysis, 17853 (63%) were genotyped. We also compared the clinical phenotype, including lung function, age at diagnosis, and nutritional measures, of 22 DeltaF508 heterozygous genotypes. Mortality rates and clinical phenotype were also compared between genotypes classified into six classes on the basis of their functional effect on CFTR production. FINDINGS: Between 1991 and 1999, genetic and clinical data were available for 17853 patients with cystic fibrosis, which was 63% of the total cohort. There were 1547 deaths during the 9 years of follow-up. In the analysis of the 11 most common genotypes, DeltaF508/R117H, DeltaF508/DeltaI507, DeltaF508/3849+10kbC-->T, and DeltaF508/2789+5G-->A had a significantly lower mortality rate (4.7, 8.0, 11.9, and 4.4, respectively) than the genotype homozygous for DeltaF508 (21.8, p=0.0060). DeltaF508/R117H, DeltaF508/DeltaI507, DeltaF508/ 3849+10 kbC-->T, DeltaF508/2789+5G-->A, and DeltaF508/A455E have a milder clinical phenotype. Outcomes for all functional classes were compared with that of class II (containing DeltaF508 homozygotes) and classes IV and V had a significantly lower mortality rate and milder clinical phenotype. INTERPRETATION: Patients with cystic fibrosis have distinct genetic subgroups that are associated with mild clinical manifestations and low mortality. These differences in phenotype are also related to the functional classification of CFTR genotype. PMID- 12767732 TI - Aortic tumour in primary anti-phospholipid syndrome. PMID- 12767733 TI - Oral topiramate for treatment of alcohol dependence: a randomised controlled trial. AB - BACKGROUND: Topiramate, a sulphamate fructopyranose derivative, might antagonise alcohol's rewarding effects associated with abuse liability by inhibiting mesocorticolimbic dopamine release via the contemporaneous facilitation of gamma amino-butyric acid activity and inhibition of glutamate function. We aimed to see whether topiramate was more effective than placebo as a treatment for alcohol dependence. METHODS: We did a double-blind randomised controlled 12-week clinical trial comparing oral topiramate and placebo for treatment of 150 individuals with alcohol dependence. Of these 150 individuals, 75 were assigned to receive topiramate (escalating dose of 25-300 mg per day) and 75 had placebo as an adjunct to weekly standardised medication compliance management. Primary efficacy variables were: self-reported drinking (drinks per day, drinks per drinking day, percentage of heavy drinking days, percentage of days abstinent) and plasma gamma glutamyl transferase, an objective index of alcohol consumption. The secondary efficacy variable was self-reported craving. FINDINGS: At study end, participants on topiramate, compared with those on placebo, had 2.88 (95% CI -4.50 to -1.27) fewer drinks per day (p=0.0006), 3.10 (-4.88 to -1.31) fewer drinks per drinking day (p=0.0009), 27.6% fewer heavy drinking days (p=0.0003), 26.2% more days abstinent (p=0.0003), and a log plasma gamma-glutamyl transferase ratio of 0.07 ( 0.11 to -0.02) less (p=0.0046). Topiramate-induced differences in craving were also significantly greater than those of placebo, of similar magnitude to the self-reported drinking changes, and highly correlated with them. INTERPRETATION: Topiramate (up to 300 mg per day) is more efficacious than placebo as an adjunct to standardised medication compliance management in treatment of alcohol dependence. PMID- 12767734 TI - Prenatal methylmercury exposure from ocean fish consumption in the Seychelles child development study. AB - INTRODUCTION: Exposure to methylmercury (MeHg) before birth can adversely affect children's neurodevelopment. The most common form of prenatal exposure is maternal fish consumption, but whether such exposure harms the fetus is unknown. We aimed to identify adverse neurodevelopmental effects in a fish-consuming population. METHODS: We investigated 779 mother-infant pairs residing in the Republic of Seychelles. Mothers reported consuming fish on average 12 meals per week. Fish in Seychelles contain much the same concentrations of MeHg as commercial ocean fish elsewhere. Prenatal MeHg exposure was determined from maternal hair growing during pregnancy. We assessed neurocognitive, language, memory, motor, perceptual-motor, and behavioural functions in children at age 9 years. The association between prenatal MeHg exposure and the primary endpoints was investigated with multiple linear regression with adjustment for covariates that affect child development. FINDINGS: Mean prenatal MeHg exposure was 6.9 parts per million (SD 4.5 ppm). Only two endpoints were associated with prenatal MeHg exposure. Increased exposure was associated with decreased performance in the grooved pegboard using the non-dominant hand in males and improved scores in the hyperactivity index of the Conner's teacher rating scale. Covariates affecting child development were appropriately associated with endpoints. INTERPRETATION: These data do not support the hypothesis that there is a neurodevelopmental risk from prenatal MeHg exposure resulting solely from ocean fish consumption. PMID- 12767735 TI - Folate treatment and unbalanced methylation and changes of allelic expression induced by hyperhomocysteinaemia in patients with uraemia. AB - BACKGROUND: Hyperhomocysteinaemia occurs in several genetically determined and acquired disorders and is highly prevalent in patients with uraemia. In these disorders, homocysteine precursor S-adenosylhomocysteine, a powerful competitive inhibitor of S-adenosylmethionine-dependent methyltransferases, is increased, suggesting unbalanced methylation. We aimed to investigate whether DNA hypomethylation is present in patients with uraemia who also have hyperhomocysteinaemia and whether regulation of specific classes of genes, dependent on DNA methylation, is compromised. METHODS: We selected men with hyperhomocysteinaemia and uraemia who were having standard haemodialysis treatment, and compared them with healthy male controls. We measured the homocysteine concentration from plasma samples and obtained DNA and RNA samples from peripheral mononuclear cells. DNA methylation was assessed by cytosine extension assay and by Southern blotting. Allelic expression of pseudoautosomal and imprinted genes was investigated by analysis of suitable restriction fragment length polymorphisms. FINDINGS: Total DNA hypomethylation was higher in patients than in controls (z score -4.593, p=0.0006) and allelic expression was changed in both sex-linked and imprinted genes. The shift from monoallelic to biallelic expression was dependent on homocysteine concentrations. Folate therapy, a common method to reduce hyperhomocysteinaemia, restored DNA methylation to normal levels and corrected the patterns of gene expression. INTERPRETATION: Our results suggest that hyperhomocysteinaemia affects epigenetic control of gene expression, which can be reverted by folate treatment. Our data support the hypothesis that the toxic action of homocysteine can be mediated by macromolecule hypomethylation. PMID- 12767736 TI - Pain in the neck. PMID- 12767737 TI - Clinical presentations and outcome of severe acute respiratory syndrome in children. AB - Hong Kong has been severely affected by severe acute respiratory syndrome (SARS). Contact in households and health-care settings is thought to be important for transmission, putting children at particular risk. Most data so far, however, have been for adults. We prospectively followed up the first ten children with SARS managed during the early phase of the epidemic in Hong Kong. All the children had been in close contact with infected adults. Persistent fever, cough, progressive radiographic changes of chest and lymphopenia were noted in all patients. The children were treated with high-dose ribavirin, oral prednisolone, or intravenous methylprednisolone, with no short-term adverse effects. Four teenagers required oxygen therapy and two needed assisted ventilation. None of the younger children required oxygen supplementation. Compared with adults and teenagers, SARS seems to have a less aggressive clinical course in younger children. PMID- 12767738 TI - Effect of gabapentin on nausea induced by chemotherapy in patients with breast cancer. AB - In an anecdotal report, complete resolution of chemotherapy-induced nausea was seen in a patient with breast cancer, after she was placed on the anticonvulsant gabapentin. On this basis, we did an open-label study in which oral gabapentin 300 mg thrice daily was given for every other chemotherapy treatment in nine patients with breast cancer. Six of the nine reported at least a three-point improvement in peak delayed nausea (on an eight-point nausea scale), and three patients had complete resolution of nausea when taking gabapentin. This preliminary evidence shows that gabapentin might have a role in treatment of chemotherapy-induced nausea. PMID- 12767739 TI - Forecasting, warning, and detection of malaria epidemics: a case study. AB - Our aim was to assess whether a combination of seasonal climate forecasts, monitoring of meteorological conditions, and early detection of cases could have helped to prevent the 2002 malaria emergency in the highlands of western Kenya. Seasonal climate forecasts did not anticipate the heavy rainfall. Rainfall data gave timely and reliable early warnings; but monthly surveillance of malaria out patients gave no effective alarm, though it did help to confirm that normal rainfall conditions in Kisii Central and Gucha led to typical resurgent outbreaks whereas exceptional rainfall in Nandi and Kericho led to true malaria epidemics. Management of malaria in the highlands, including improved planning for the annual resurgent outbreak, augmented by simple central nationwide early warning, represents a feasible strategy for increasing epidemic preparedness in Kenya. PMID- 12767740 TI - Brundtland meets food and drink leaders but declines Coke cocktail. PMID- 12767741 TI - China takes drastic action over SARS threat. PMID- 12767742 TI - Australia shaken by complementary medicines recall. PMID- 12767744 TI - HIV prevention efforts reach fewer than one in five at risk. PMID- 12767746 TI - Therapy found for Parkinson's disease dyskinesia. PMID- 12767748 TI - AIDS fund short of money. PMID- 12767749 TI - Brazil faces worst outbreak of conjunctivitis in 20 years. PMID- 12767750 TI - Melioidosis. AB - Melioidosis, which is infection with the gram-negative bacterium Burkholderia pseudomallei, is an important cause of sepsis in east Asia and northern Australia. In northeastern Thailand, melioidosis accounts for 20% of all community-acquired septicaemias, and causes death in 40% of treated patients. B pseudomallei is an environmental saprophyte found in wet soils. It mostly infects adults with an underlying predisposing condition, mainly diabetes mellitus. Melioidosis is characterised by formation of abscesses, especially in the lungs, liver, spleen, skeletal muscle, and prostate. In a third of paediatric cases in southeast Asia, the disease presents as parotid abscess. In northern Australia, 4% of patients present with brain stem encephalitis. Ceftazidime is the treatment of choice for severe melioidosis, but response to high dose parenteral treatment is slow (median time to abatement of fever 9 days). Maintenance antibiotic treatment is with a four-drug regimen of chloramphenicol, doxycycline, and trimethoprim-sulfamethoxazole, or with amoxicillin-clavulanate in children and pregnant women. However, even with 20 weeks' antibiotic treatment, 10% of patients relapse. With improvements in health care and diagnostic microbiology in endemic areas of Asia, and increased travel, melioidosis will probably be recognised increasingly during the next decade. PMID- 12767751 TI - 25 years of the WHO essential medicines lists: progress and challenges. AB - The first WHO essential drugs list, published in 1977, was described as a peaceful revolution in international public health. The list helped to establish the principle that some medicines were more useful than others and that essential medicines were often inaccessible to many populations. Since then, the essential medicines list (EML) has increased in size; defining an essential medicine has moved from an experience to an evidence-based process, including criteria such as public-health relevance, efficacy, safety, and cost-effectiveness. High priced medicines such as antiretrovirals are now included. Differences exist between the WHO model EML and national EMLs since countries face varying challenges relating to costs, drug effectiveness, morbidity patterns, and rationality of prescribing. Ensuring equitable access to and rational use of essential medicines has been promoted through WHO's revised drug strategy. This approach has required an engagement by WHO on issues such as the effect of international trade agreements on access to essential medicines and research and development to ensure availability of new essential medicines. PMID- 12767752 TI - A multicentre collaboration to investigate the cause of severe acute respiratory syndrome. AB - Severe acute respiratory syndrome is a new disease in human beings, first recognised in late February, 2003, in Hanoi, Vietnam. The severity of the disease, combined with its rapid spread along international air-travel routes, prompted WHO to set up a network of scientists from 11 laboratories around the world to try to identify the causal agent and develop a diagnostic test. The network unites laboratories with different methods and capacities to rapidly fulfil all postulates for establishing a virus as the cause of a disease. Results are shared in real time via a secure website, on which microscopy pictures, protocols for testing, and PCR primer sequences are also posted. Findings are discussed in daily teleconferences. Progress is further facilitated through sharing between laboratories of samples and test materials. The network has identified a new coronavirus, consistently detected in samples of SARS patients from several countries, and conclusively named it as the causative agent of SARS; the strain is unlike any other known member of the genus Coronavirus. Three diagnostic tests are now available, but all have limitations. PMID- 12767753 TI - UK Biobank: a project in search of a protocol? AB - Before it has even begun recruiting participants, the UK Biobank project has raised extraordinary passions among scientists. Some scientists are broadly supportive of the project, and feel that it is a scientifically valid, potentially valuable resource, although they still have reservations over the details of the current protocol. Others see it as an ill-conceived, politically motivated project, in which consultations have only been done to give an appearance of legitimacy and in which the scientific case has not been made for its design. There has been resentment over how the protocol has evolved, and the secrecy and legal constraints that surrounded the process for groups bidding to be involved has had the appearance of attempting to stifle debate. For this article even supporters of the project have been unwilling to be quoted on the record. The only point that everyone seems to agree on is that sufficient debate about the project has not taken place. PMID- 12767754 TI - SARS, lay epidemiology, and fear. PMID- 12767757 TI - Admission cardiotocography. PMID- 12767755 TI - Atypical presentations of SARS. PMID- 12767758 TI - Admission cardiotocography. PMID- 12767759 TI - Hepatic artery chemotherapy and colorectal liver metastases. PMID- 12767760 TI - Hepatic artery chemotherapy and colorectal liver mestastases. PMID- 12767761 TI - Hepatic artery chemotherapy and colorectal liver mestastases. PMID- 12767763 TI - Does VaxGen hide the breakthrough infections? PMID- 12767764 TI - Inequities among the very poor: effect of young age on care-seeking. PMID- 12767765 TI - Recombinant activated factor VII and perioperative blood loss. PMID- 12767766 TI - Recombinant activated factor VII and perioperative blood loss. PMID- 12767768 TI - Churg-Strauss syndrome. PMID- 12767769 TI - Cancer patients' anxiety. PMID- 12767770 TI - Mortality, morbidity, and injury of children living with single parents. PMID- 12767771 TI - Mortality in parents after death of a child. PMID- 12767772 TI - Mortality, morbidity, and injury of children living with single parents. PMID- 12767774 TI - Staining for cell death. PMID- 12767776 TI - Private medical education takes off in India. PMID- 12767777 TI - Primary-care research is not a lost cause. PMID- 12767778 TI - Randomised controlled trials and quality of journals. PMID- 12767780 TI - Importance of language. PMID- 12767781 TI - Importance of language. PMID- 12767782 TI - Evidence-based programme design for CME. PMID- 12767784 TI - Neurasthenia. PMID- 12767787 TI - Current concepts of the inflammatory response after major trauma: an update. AB - During the past century explosive developments have taken place in the field of molecular medicine and genetics, vastly expanding our understanding of the normal physiological response to injury. We have been able to characterise specific molecular and cell biological processes and apply some of this knowledge to the treatment of multiply injured patients. Despite the significant steps we have made, there still remains much work to be done in this area. This review article highlights the current concepts of post-traumatic immunological changes and their impact in the management of trauma patients. PMID- 12767788 TI - Thoracolumbar fracture in blunt trauma patients: guidelines for diagnosis and imaging. AB - OBJECTIVE: The aim of the study was to determine a clinical diagnostic pathway for the imaging of the thoracolumbar spine in blunt trauma patients. METHOD: A literature review was undertaken to determine the factors important in the detection of thoracolumbar injury and also to produce a trial protocol, which could be tested retrospectively. Two hundred patients admitted between 1998 and 2000 were reviewed retrospectively to form a database of the relevant clinical features. The diagnostic pathway protocol was tested retrospectively in two groups; one hundred with known thoracolumbar fractures and one hundred undifferentiated multi-trauma patients. Sensitivity and specificity indices were determined using the chi-squared test of association. RESULTS: Falls and motor vehicle crashes were the most common mechanism of injury causing thoracolumbar fractures. No significant association could be found between mechanism of injury (MOI) and presence of thoracolumbar fracture. Clinical signs found to be associated with thoracolumbar fracture include: (1) back pain/midline tenderness (Sens 62.1% Spec 91.5%), (2) palpable midline step (Sens 13.8% Spec 100%), (3) back bruising (Sens 6.9% Spec 98.6%), (4) abnormal neurological signs (Sens 41.4% Spec 95.8%). Cervical level fractures were also found to be associated with an increased incidence of thoracolumbar fractures. Factors found to influence the detection of back pain/midline tenderness include: (1) GCS<15, (2) ETOH/drug intoxication, (3) major distracting injury. The proposed diagnostic pathway would have led to 92% of patients receiving TL imaging in group 2 (multi-trauma) with a sensitivity of 100%, specificity of 11.3% and a negative predictive value of 100%. CONCLUSION: From the literature and the results of the study, we propose that imaging of the thoracolumbar spine is required in those patients suffering from a high force mechanism of injury if any of the following are present: (1) back pain/midline tenderness, (2) local signs of thoracolumbar injury, (3) abnormal neurological signs, (4) cervical spine fracture, (5) GCS<15, (6) major distracting injury, (7) ETOH/ drug intoxication. An evidence based diagnostic pathway has been shown to be highly sensitive in determining the presence of TL fracture in a retrospective study of blunt trauma patients. PMID- 12767789 TI - Physical and psychiatric predictors of late whiplash syndrome. AB - Approximately 40% of people who sustain whiplash injuries, complain of chronic physical and psychiatric symptoms. Associations have been found between pre accident physical symptoms and physical outcome and between pre-accident psychiatric variables and both physical and psychiatric outcome. There are no reported investigations of the association between pre-accident physical symptoms and psychiatric outcome. In this study, 33 consecutive cases of whiplash injury met inclusion criteria from a series of psychiatric reports used in civil litigation. Outcome was measured for each patient and correlated with a range of pre-accident physical and psychiatric variables using multivariate regression. There was no association between pre-accident psychiatric factors and overall outcome. Older age and a pre-accident history of musculoskeletal complaints correlated with the physical and psychiatric outcome. In whiplash injury, pre accident psychiatric factors may have little bearing on long-term prognosis. Physical and psychiatric outcome of late whiplash syndrome is probably worse in older individuals and in patients with a pre-accident history of musculoskeletal complaints. PMID- 12767790 TI - Acromioclavicular joint sprains: the post-injury recovery interval. AB - Acromioclavicular joint (ACJ) sprains occur after injuries to the shoulder girdle. Current practice is to treat such injuries conservatively. This study determines the recovery interval of ACJ sprains treated conservatively. Forty seven patients with grades I or II ACJ sprains were evaluated in a shoulder clinic and treated conservatively. The average age was 37 years, with the injury being caused by a fall, contact sport or road-traffic accident. At an interval of 12 months or greater these patients were assessed by means of a questionnaire. Six months post-injury, pain was described as significant by 14 (40%) patients, decreasing to 5 patients (14%) at final follow-up. A restricted range of movement was reported by seven patients (20%) 6 months post-injury. There is a positive correlation between patients symptomatic at 6 months and those whose symptoms persist beyond 1 year (r=0.6, 95% CI=0.28-0.76, P<0.01). Three patients (9%) found that symptoms affected their ability to perform activities of daily living and two patients (6%) had to change sporting activities. Patients may continue to experience adverse symptoms beyond 6 months and should be advised accordingly on appropriate treatment should symptoms persist. PMID- 12767791 TI - Least possible fixation of fractures of the proximal humerus. AB - The purpose of this study was to assess the clinical outcome of an unreported technique of the least possible fixation for valgus-impacted fractures. Using an open technique without any sharp deep soft tissue dissection, the split between the tuberosities is opened to allow elevation of the impacted humeral head fragment. The tuberosities are reduced and repaired with absorbable sutures between the rotator cuff insertions. No bone grafting, Kirschner wire or other fixation device was used. For 11 patients, with an average age of 55 years, who were operated between 1989 and 1998, the mean follow-up period was 69 months. The patients were assessed radiologically and clinically using the Constant-Murley score. As a percentage of the normal side, the Constant-Murley scores ranged from 55 to 100% (mean 86%); there were eight patients with scores >80%, two patients with scores between 70 and 79% and one patient with score <70%. One patient (9%) showed avascular necrosis (AVN). This fixation allows preservation of the soft tissues in an open approach and avoids the dangers of other fixation methods. There is a low risk of AVN and it allows a good functional result. PMID- 12767792 TI - Posterior sternoclavicular joint dislocation: the value of intra-operative ultrasound. AB - The sternoclavicular joint dislocates posteriorly if the costoclavicular ligaments, posterior capsule, and anterior capsule have all been disrupted. Diagnosis of a posteriorly dislocated sternoclavicular joint is difficult on clinical examination and with plain X-rays. The treatment objective is to obtain a closed reduction, and failing this an open reduction is indicated. In the intra operative setting the evaluation of the reduction when performed through closed means can be somewhat difficult, and intra-operative plain films do not give clear answers. Ultrasound as an imaging modality for these injuries was shown to demonstrate the state of the joint in terms of reduction, more clearly than X rays (P<0.001). It was accurately interpreted by the vast majority of orthopaedic surgeons in an investigation, and is of great value in the intra-operative setting to confirm whether a closed reduction has been successful or not. A case is reported illustrating its use. PMID- 12767793 TI - Calcaneal fractures in adolescents. CT classification and results of operative treatment. AB - The morphology of calcaneal fractures in 9 adolescents (mean age 13.4 years) with 10 fractures were classified using plain films and computed tomography scans. The patterns were found to be similar to those in adults. All except one of the fractures (which was not significantly displaced) were treated with open reduction and internal fixation. In all cases it was possible to achieve anatomic reduction and rigid internal fixation. Seven patients had 'excellent' long-term clinical results. One patient with pending litigation scored 'good', and one patient with an ipsilateral fracture of the talar neck scored 'fair'. This patient had mild limitation of ankle movement, all others had full ankle movement. Five had unrestricted subtalar movement, in two it was mildly limited and in three it was moderately limited (50-80%). There was no evidence of abnormality of the physes on follow up X-rays. We conclude that operative treatment of this fracture yields good results. PMID- 12767794 TI - Selected Abstracts from the Southern Trauma Symposium and Australian Trauma Society Meeting, held on the 8 November 2002. PMID- 12767795 TI - Sternoclavicular hyperostosis with pathological fracture of the clavicle--a case report. PMID- 12767796 TI - Unusual fracture dislocation of the distal end of clavicle in an adolescent. PMID- 12767797 TI - Orthopaedic variant of Munchausen's syndrome: shoulder instability. PMID- 12767798 TI - Fracture-dislocation of the first metatarsophalangeal joint Clark et al. [Injury 31 (2000) 465-466]. Is this dislocation plantar, Jahss IV dorsal or pathological? PMID- 12767799 TI - The operative management of displaced intra-articular fractures of the calcaneum: a two-centre study using a defined protocol. PMID- 12767800 TI - The use of subtalar arthroscopy in open reduction and internal fixation of intra articular calcaneal fractures. PMID- 12767802 TI - Production of the bullous pemphigoid antigen 230 (BP230) by Saccharomyces cerevisiae and Pichia pastoris. AB - BP230 is a cytoskeletal linker protein of 2649 amino acids originally identified as the target autoantigen in bullous pemphigoid, a potentially devastating autoimmune skin blistering disorder. To better define its function, we sought to generate recombinant forms of BP230 in both Saccharomyces cerevisiae and Pichia pastoris after cloning its entire cDNA. By immunoblot analysis, full-length BP230 was not found in extracts of P. pastoris, whereas minor amounts of degraded BP230 were detected in extracts of S. cerevisiae. In contrast, both S. cerevisiae and P. pastoris were able to produce the 770-amino acid COOH-terminal domain of BP230. Furthermore, the production level of the recombinant BP230 tail in S. cerevisiae was significantly higher than that observed in P. pastoris and that of endogenous BP230 in cultured human keratinocytes. Finally, 12 of 17 (71%) BP sera recognized the recombinant BP230 protein in yeast extracts. Our results indicate that S. cerevisiae occasionally constitutes a better tool for recombinant protein production than P. pastoris. Although both its large size and poor solubility limit production of BP230, the developed yeast system provides cellular fractions enriched in BP230 recombinant proteins that constitute useful tools for the diagnosis of bullous pemphigoid. PMID- 12767803 TI - Expression, purification, and biochemical characterization of Chk, a soluble protein tyrosine kinase. AB - CSK family contains two protein tyrosine kinases: Csk (C-terminal Src kinase) and Chk (Csk homologous kinase). They are responsible for phosphorylating Src family protein tyrosine kinases on a C-terminal Tyr (Tyr527) and negatively regulating their activities. However, Chk and Csk have different expression patterns, mechanisms of regulation, and different biological functions, and appear to play different roles in the development of breast cancer. To obtain pure human Chk for biochemical characterization, its coding region was amplified by polymerase chain reaction and expressed as a fusion protein with glutathione S-transferase in Escherichia coli. The enzyme was highly expressed but unusually prone to proteolytic degradation during purification. Expression of the enzyme as a dual fusion protein with glutathione S-transferase on N-terminus and streptag, a 10 amino acid peptide, on C-terminus allowed purification of the full-length fusion protein. The purified enzyme was able to phosphorylate and inactivate Src. Chk (no inhibition up to 18.5 microM) and Csk (IC(50)= 1 microM) were differentially inhibited by PP2, probably due to the size difference of one residue (Thr265 in Csk versus Met304 in Chk) in the ATP-binding domain. The expression, purification, and initial characterizations of Chk provided an important step toward full characterization of Chk and Csk, two important enzymes in cellular regulation. PMID- 12767804 TI - Isolation of a novel N-acetylglucosamine-specific lectin from fresh sclerotia of the edible mushroom Pleurotus tuber-regium. AB - An N-acetylglucosamine-binding lectin with a molecular mass of 32kDa was isolated from fresh sclerotia of the edible mushroom Pleurotus tuber-regium. Its N terminal sequence exhibited some similarity to that of Agaricus bisporus lectin. The isolation procedure was simple, involving (NH(4))(2)SO(4) precipitation, ion exchange chromatography on DEAE-cellulose, affinity chromatography on N-acetyl-D glucosamine-agarose, and gel filtration by fast protein liquid chromatography on Superdex 75. The lectin exhibited hemagglutinating activity toward trypsinized rabbit erythrocytes but not toward untrypsinized rabbit erythrocytes. PMID- 12767805 TI - Overexpression of a small medicinal peptide from ginseng in the yeast Pichia pastoris. AB - A medicinal peptide, Gsp, which was initially extracted from the traditional medicinal herb ginseng, has potential use as a drug against diabetes. Gsp is a low molecular weight protein that we have secreted in a recombinant form from the yeast Pichia pastoris. A DNA fragment encoding four copies of the Gsp protein each separated by a basic amino acid was synthesized and inserted into the P. pastoris expression vector plasmid pPIC9. After electroporation of the resulting vector, pPIC9-Gsp, into the yeast, transformants were selected. Recombinant pre Gsp secreted from P. pastoris had a molecular weight of 5.9 kDa and mature recombinant Gsp had a primary structure indistinguishable from native Gsp. After optimization of the culturing process, the yield of pre-Gsp reached 800 mg/L in the clarified broth. A continuous batch fermentation process was developed that allowed the same population of cells to be reutilized five times without loss of expression level. This continuous culturing process resulted in a substantial saving of both time and cost in pharmaceutical production and should be applicable to the production of other recombinant proteins in P. pastoris. PMID- 12767806 TI - Construction of a modified vector for efficient purification of recombinant Mycobacterium tuberculosis proteins expressed in Escherichia coli. AB - A major problem in assessing the vaccine and diagnostic potential of various proteins encoded by Mycobacterium tuberculosis genome is the inability to produce large quantities of these proteins, even when Escherichia coli or other heterologous systems are employed for recombinant protein production. To overcome these barriers, we have constructed a modified expression vector, using pGEX-4T-1 vector as the backbone. In addition to the features offered by the pGEX-4T vectors, the new vector allowed easy purification of recombinant proteins on the highly versatile Ni-NTA-agarose affinity matrix. The utility of the new vector was demonstrated by expressing and purifying, to near homogeneity, two M. tuberculosis proteins, i.e., Rv3872 (a member of the multi-gene PE subfamily) and Rv3873 (a member of the multi-gene PPE subfamily), which are encoded by the RD1 region of M. tuberculosis. The proteins encoded by rv3872 and rv3873 were expressed at high levels as fusion proteins with glutathione-S-transferase in E. coli. The recombinant Rv3872 and Rv3873 proteins were purified and isolated free of the fusion partner (GST) by affinity purification on glutathione-Sepharose and/or Ni-NTA-agarose affinity matrix and cleavage of the purified fusion proteins by thrombin protease. The recombinant Rv3872 protein was nearly homogeneous (more than 95% pure) while Rv3873 preparation was more than 90% pure. The recombinant Rv3872 and Rv3873 proteins were immunologically active and reacted with antibodies in sera from TB patients. Our results demonstrate the utility of the newly constructed expression vector with two affinity tags for efficient expression and purification of recombinant M. tuberculosis proteins expressed in E. coli, which could be used for further diagnostic and immunological studies. PMID- 12767807 TI - Production and characterization of the Talaromyces stipitatus feruloyl esterase FAEC in Pichia pastoris: identification of the nucleophilic serine. AB - Feruloyl esterases constitute an interesting group of enzymes that have the potential for use over a broad range of applications in the agri-food industries. We report the over-expression and characterization of a novel feruloyl esterase exhibiting broad substrate specificity from Talaromyces stipitatus (FAEC) in Pichia pastoris. Using various gene constructions, we have investigated the use of alternative signal peptides to produce an authentic feruloyl esterase featuring the N-terminal sequence determined for the native enzyme. We demonstrate that additional amino acids at the N-terminus of the FAEC sequence do not influence the catalytic capacity of the enzyme, and that the nature of the signal sequence has a limited effect on the yield of the secreted enzyme, with the T. stipitatus FAEC signal sequence producing 297 mgL(-1), the Neurospora crassa Fae-1 260 mgL(-1), and the Saccharomyces cerevisiae alpha-factor secretion signal 214 mgL(-1). Mature FAEC contains two internal peptide sequences that correspond with the consensus motif G-X-S-X-G that contains the catalytic serine nucleophile, which is conserved in the esterase enzyme superfamily. The serine residues at the center of these peptide motifs have been independently mutated and the corresponding enzymes have been over-expressed in P. pastoris to identify the candidate nucleophilic residue responsible for catalyzing the enzymatic reaction. Purified recombinant FAEC containing S465A retained the esterase activity and appeared unaffected by the amino acid modification. In contrast, FAEC activity containing S166A was below the HPLC detection limit, suggesting that serine 166 constitutes the nucleophile. PMID- 12767808 TI - The first semi-synthetic serine protease made by native chemical ligation. AB - Selective incorporation of non-natural amino acid residues into proteins is a powerful approach to delineate structure-function relationships. Although many methodologies are available for chemistry-based protein engineering, more facile methods are needed to make this approach suitable for routine laboratory practice. Here, we describe a new strategy and provide a proof of concept for engineering semi-synthetic proteins. We chose a serine protease Streptomyces griseus trypsin (SGT) for this study to show that it is possible to efficiently couple a synthetic peptide containing a catalytically critical residue to a recombinant fragment containing the other active site residues. The 223-residue hybrid SGT molecule was prepared by fusing a chemically synthesized N-terminal peptide to a large C-terminal fragment of recombinant origin using native chemical ligation. This C-terminal polypeptide was produced from full-length SGT by cyanogen bromide cleavage at a genetically engineered Met57 position. This semi-synthetic hybrid trypsin is fully active, showing kinetics identical to the wild-type enzyme. Thus, we believe that it is an ideal model enzyme for studying the catalytic mechanisms of serine proteases by providing a straightforward approach to incorporate non-natural amino acids in the N-terminal region of the protein. In particular, this strategy will allow for replacement of the catalytic His57 residue and the buried N-terminus, which is thought to help align the active site, with synthetic analogs. Our approach relies on readily available recombinant proteins and small synthetic peptides, thus having general applications in chemical engineering of large proteins where the N-terminal region is the focal interest. PMID- 12767809 TI - Expression of a Brassic napus glutamate 1-semialdehyde aminotransferase in Escherichia coli and characterization of the recombinant protein. AB - Glutamate 1-semialdehyde aminotransferase (GSA-AT) is a key regulatory enzyme, which converts glutamate 1-semialdehyde (GSA) to 5-aminolevulinic acid (ALA) in chlorophyll biosynthesis. ALA is the universal precursor for the synthesis of chlorophyll, heme, and other tetrapyrroles. To study the regulation of chlorophyll biosynthesis in Brassica napus, two cDNA clones of GSA-AT were isolated for genetic manipulation. A SalI-XbaI fragment from one of the two cDNA clones of GSA-AT was used for recombinant protein expression by inserting it at the 3' end of a calmodulin-binding-peptide (CBP) tag of the pCaln vector. The CBP tagged recombinant protein, expressed in Escherichia coli, was purified to apparent homogeneity in a one step purification process using a calmodulin affinity column. The purified CBP tagged GSA-AT is biologically active and has a specific activity of 16.6 nmol/min/mg. Cleavage of the CBP tag from the recombinant protein with thrombin resulted in 9.2% loss of specific activity. However, removal of the cleaved CBP tag from the recombinant protein solution resulted in 60% loss of specific activity, suggesting possible interactions between the recombinant protein and the CBP tag. The enzyme activity of the CBP tagless recombinant protein, referred as TR-GSA-AT hereafter, was not affected by the addition of pyridoxamine 5' phosphate (PMP). Addition of glutamate and pyridoxal 5' phosphate (PLP) to the TR-GSA-AT enhanced the enzyme activity by 3 fold and 3.6-fold, respectively. Addition of both glutamate and PLP increased the enzyme activity by 4.6-fold. Similar to the GSA-AT of B. napus, the active TR-GSA AT is a dimeric protein of 88 kDa with 45.5 kDa subunits. As the SalI-XbaI fragment encodes a biologically active GSA-AT that has the same molecular mass as the native GSA-AT, it is concluded that the SalI-XbaI fragment is the coding sequence of GSA-AT. The highly active polyclonal antibodies generated from TR-GSA AT were used for the detection of GSA-AT of B. napus. PMID- 12767810 TI - Isolation of alpha-lactalbumin, beta-lactoglobulin, and bovine serum albumin from cow's milk using gel filtration and anion-exchange chromatography including evaluation of their antigenicity. AB - The aim of this study was to introduce a simple, reproducible, and less expensive method for isolation of alpha-lactalbumin, beta-lactoglobulin, and bovine serum albumin from cow's milk while retaining their antigenicity. Whey (lactoserum) was obtained by isolating casein from defatted milk using hydrochloric acid. Globulins were then precipitated from whey by half-saturated ammonium sulfate and beta-lactoglobulin was purified further using Sephadex G-50 gel filtration. The proteins in the supernatant were also fractionated using diethylaminoethyl cellulose chromatography in which beta-lactoglobulin was separated from alpha lactalbumin and bovine serum albumin. The latter two proteins that co-eluted in anion-exchange chromatography were then gently isolated from each other by Sephadex G-50 gel filtration. Pure beta-lactoglobulin was also obtained by anion exchange chromatography of the ammonium sulfate-precipitated globulins. Using enzyme-linked immunosorbent assay (ELISA), Western blotting, and ELISA inhibition assay, antigenicity of the purified proteins was evaluated. Our results showed high purity and well-preserved antigenicity of alpha-lactalbumin, beta lactoglobulin, and bovine serum albumin thus purified. PMID- 12767811 TI - Purification and folding of recombinant bovine oxoglutarate/malate carrier by immobilized metal-ion affinity chromatography. AB - A major obstacle to investigating the structure of membrane proteins is the difficulty in obtaining sufficient amounts of functional protein. The oxoglutarate carrier, an intrinsic membrane-transport protein of the inner membranes of bovine-heart mitochondria, has been cloned as a fusion protein containing a C-terminal hexa-histidine tag. This fusion protein has been expressed at an abundant level in a mutant strain of Escherichia coli BL21 (DE3) called C41 (DE3). The protein accumulated as inclusion bodies and none was detected in the bacterial inner membrane. The denatured protein was refolded to reconstitute functional properties similar to the native protein. Solubilized inclusion body protein was immobilized using nickel-chelating affinity chromatography, and purified and refolded in a single step. The protein eluted as a monomer which was stable in mild detergent, at a yield equivalent to 15 mg active protein/liter bacterial culture. The reconstituted fusion protein displayed the same transport characteristics as the wild-type, demonstrating that the tag does not perturb the structure of the protein. The oxoglutarate carrier is one member of an extensive family of mitochondrial transport proteins. These proteins transport many different metabolites across the inner mitochondrial membrane and share a common mechanism of action. Therefore, it is likely that this folding protocol can be applied successfully to other mitochondrial transport proteins, thus providing sufficient protein for extensive crystallization trials with a wide range of family members. PMID- 12767812 TI - Purification of Her-2 extracellular domain and identification of its cleavage site. AB - The EGF family of receptors belongs to the tyrosine kinase receptor (TKR) family and plays an important role during embryonic and postnatal development and also in the progression of tumors. Her-2/neu/c-erbB-2, a member of the epidermal growth factor receptor family, can be cleaved into a soluble extra cellular domain (ECD) and a membrane-bound stub fragment. Her-2 ECD from a breast cancer cell line SKBR3 was immunopurified and analyzed with matrix-assisted laser desorption ionization (MALDI) and carboxyl terminal amino acid sequencing. A sequence within the juxtamembrane region (only 11 amino acid residues) PAEQR ASP was identified most likely as a primary site of cleavage, PA EQRASP as a minor site, that generate the ECD. The sites of cleavage are within the signature motif P/GX(5-7)P/G highly conserved in the EGF receptor family. PMID- 12767813 TI - High-level expression, secretion, and purification of the thermostable aqualysin I from Thermus aquaticus YT-1 in Pichia pastoris. AB - Aqualysin I is a heat-stable subtilisin-type serine protease which is secreted into the culture medium by Thermus aquaticus YT-1, an extreme thermophile. We report the high-level expression of an aqualysin I protein using its native signal sequence for secretion in the methylotrophic yeast, Pichia pastoris. The expression of aqualysin I in P. pastoris was carried out using the glyceraldehyde 3-phosphate dehydrogenase (GAP) promoter. Pro-aqualysin I (38kDa) as inactive protein was secreted into the medium of shake-flask cultures at a concentration of 1g/L. It was isolated from the culture supernatant by an ammonium sulfate precipitation and one-step anion exchange chromatography in a nearly pure form and was autoproteolytically activiated by heat treatment. A proteolytic activity test indicated that the purified recombinant aqualysin I was properly folded with a specific activity similar to that of the native enzyme. We also explored the possibility of secreting the GAP expressed aqualysin I in P. pastoris by in-frame fusion of the Saccharomyces cerevisiae alpha-factor secretion signal. However, the levels of secreted pro-aqualysin I particles were approximately 10 times lower, possibly as a consequence of membrane association or to the influence of the alpha-factor secretion signal sequences on the transcription or secretion of aqualysin I. When considering further optimization of the downstream process and culture conditions for high-level production of recombinant aqualysin I by P. pastoris, this expression system is promising for development as an industrial process. PMID- 12767814 TI - Expression, purification, and characterization of arginine kinase from the sea cucumber Stichopus japonicus. AB - The arginine kinase gene of sea cucumber Stichopus japonicus was cloned and inserted into the prokaryotic expression plasmid pET-21b. The protein was expressed in a soluble and functional form in Escherichia coli and purified by Blue Sepharose CL-6B, DEAE-32, and Sephadex G-100 chromotography with a final yield of 83 mgL(-1) of LB medium. The specific activity, electrophoretic mobility, and isoelectric focusing were all identical with those of arginine kinase that was purified from sea cucumber muscle. The fluorescence emission spectrum of arginine kinase had a maximum fluorescence at a wavelength of 330 nm upon excitation at 295 nm. These results are the first report of this purified protein. PMID- 12767815 TI - Expression and in vitro activation of Manduca sexta prophenoloxidase-activating proteinase-2 precursor (proPAP-2) from baculovirus-infected insect cells. AB - Prophenoloxidase activation is a component of the immune system in insects and crustaceans. We recently purified and cloned a new prophenoloxidase-activating proteinase (PAP-2) from hemolymph of the tobacco hornworm Manduca sexta [J. Biol. Chem. 278, 3552-3561]. As the terminal component of a putative serine proteinase cascade, this enzyme activates prophenoloxidase (proPO) via limited proteolysis. To purify and study the activating proteinase for PAP-2 from this insect, we expressed the zymogen of PAP-2 (proPAP-2) in insect cells infected by a recombinant baculovirus that harbors the cDNA. To facilitate the purification of proPAP-2, we modified a commercial vector (pFastBac1) by inserting a synthetic DNA fragment encoding a hexahistidine sequence, allowing fusion of the affinity tag to the carboxyl terminus of a protein. After Spodoptera frugiperda Sf21 cells were infected by the virus, recombinant proPAP-2 was efficiently secreted into the media at a concentration of 5.9 microg/ml under the optimal conditions. After ammonium sulfate precipitation, the proenzyme was purified to near homogeneity by affinity chromatography on Ni(2+)-NTA agarose. Western blot analysis indicated that the recombinant proPAP-2 has a mobility slightly lower than that of the zymogen from M. sexta hemolymph. The molecular mass and isoelectric point of proPAP-2 were determined to be 47,573+/-11Da and 6.6, respectively. After the purified proenzyme was added to hemolymph from induced M. sexta larvae, it was rapidly activated by an unknown proteinase in the presence of peptidoglycan. PMID- 12767816 TI - Expression of Pseudomonas stutzeri Zobell cytochrome c-551 and its H47A variant in Escherichia coli. AB - The nirM gene encoding cytochrome c-551 from Pseudomonas stutzeri Zobell (PZ) has been expressed in Escherichia coli at levels higher than those previously reported but only under strict anaerobic growth conditions. Expression yields for wild-type cytochrome in this study typically reached 0.6 micromol per liter of saturated E. coli culture (5.5mg/L). Culture conditions investigated are compared to obtained c-551 expression levels; the results may lead to a greater understanding of the challenges encountered when expressing c-type hemoproteins in E. coli. The nirM gene was mutated to produce a histidine-47-alanine mutation of c-551 that been heterologously expressed in E. coli using optimum culture conditions and had its physiochemical properties compared to those of the wild type protein. In PZ, the histidine-47 residue is part of a conserved hydrogen bonding network located at the bottom of the heme crevice that also involves tryptophan-56 and a heme propionate. Ionization events within this network are experimentally demonstrated to modulate c-551 oxidation-reduction potential and its observed dependence on pH around neutrality. The redox potential of the mutant cytochrome still displays pH-dependence; however, the midpoint potential is approximately 25mV lower with respect to wild-type c-551 at neutral pH while the pK at which the heme propionate (HP-17) ionizes is lowered by 1.3 pH units. Temperature and chemical denaturant studies also show that loss of the hydrogen bond-donating imidazole leads to a large decrease in c-551 tertiary stability. PMID- 12767817 TI - Overexpression and purification of isotopically labeled Escherichia coli MutH for NMR studies. AB - MutH is one of the enzymes involved in the methyl directed -GATC-based DNA repair system. We report a significantly optimized protocol to prepare isotopically (15N and/or 13C) labeled MutH in minimal medium with high yields for NMR studies. Under the various conditions that we have standardized for the affinity purification of His(6) MutH, the yield of the purified MutH has been estimated to be 35-40 mg of protein from 1liter of M9 minimal media. The yield, thus, obtained by this method is significantly higher than those of previously reported methods. Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectroscopy analysis revealed that the protein was pure and existed essentially in a monomeric form. Uniformly 15N-labeled protein, thus, produced has been characterized by recording a sensitivity enhanced 2D [15N]-[1H] HSQC spectrum. The dispersion seen in 15N-1H cross-peaks indicates the presence of a well ordered structure for MutH and proper folding of the purified protein. The spectrum confirms further the existence of MutH as a monomer. PMID- 12767819 TI - Recombinant expression of Mus musculus myoglobin. AB - The cDNA encoding for Mus musculus myoglobin (Mb) was amplified using standard RT PCR techniques and cloned in an appropriate bacterial expression vector. For the first time, mouse Mb was recombinantly expressed in Escherichia coli cells, BL21(DE3), and purified in sufficient amounts to carry out a preliminary characterization. As shown by mass spectrometry, the protein is found in complex with glutathione, which binds the Cys residue in the topological position E9, in the proximity of the heme group. In recombinant murine Mb, azide affinities are only slightly dependent on the Cys(E9) oxidation state. This suggests that Cys(E9) does not provide a relevant contribution for the stabilization of ligands bound to the heme iron atom. Recombinant expression of M. musculus Mb might have an important role in order to investigate the eventual involvement of Cys(E9) in the new physiological roles proposed for Mb. PMID- 12767818 TI - DNA gyrase and DNA topoisomerase of Bacillus subtilis: expression and characterization of recombinant enzymes encoded by the gyrA, gyrB and parC, parE genes. AB - Bacillus subtilis Bs gyrA and gyrB genes specifying the DNA gyrase subunits, and parC and parE genes specifying the DNA topoisomerase IV subunits, have been separately cloned and expressed in Escherichia coli as hexahistidine (his6) tagged recombinant proteins. Purification of the gyrA and gyrB subunits together resulted in predominantly two bands at molecular weights of 94 and 73kDa; purification of the parC and parE subunits together resulted in predominantly two bands at molecular weights of 93 and 75kDa, as predicted by their respective sequences. The ability of the subunits to complement their partner was tested in an ATP-dependent decatenation/supercoiling assay system. The results demonstrated that the DNA gyrase and the topoisomerase IV subunits produce the expected supercoiled DNA and relaxed DNA products, respectively. Additionally, inhibition of these two enzymes by fluoroquinolones has been shown to be comparable to those of the DNA gyrases and topoisomerases of other bacterial strains. In sum, the biological and enzymatic properties of these products are consistent with their authenticity as DNA gyrase and DNA topoisomerase IV enzymes from B. subtilis. PMID- 12767820 TI - Expression of Sonic hedgehog-Fc fusion protein in Pichia pastoris. Identification and control of post-translational, chemical, and proteolytic modifications. AB - We have investigated the suitability of Pichia pastoris as an expression system for the candidate therapeutic protein, Sonic hedgehog fused to an immunoglobulin Fc domain (Shh-Fc). Sonic hedgehog is a morphogen protein involved in the patterning of a wide range of tissues during animal embryogenesis. The presence of Sonic hedgehog and its receptor, Patched, in adult nervous tissue suggests possible applications for the protein in the treatment of neurodegenerative disease and injury. We have engineered the Shh-Fc fusion protein in order to improve binding affinity and increase systemic exposure in animals. N-terminal sequencing, peptide mapping, mass spectrometry, and other biochemical and biological methods were used to characterize the purified protein. These analyses revealed several unanticipated problems, including thiaproline modification of the N-terminal cysteine, cleavage by a Kex2-like protease at a site near the N terminus, proteolysis at sites near the hinge, addition of a hexose in the CH3 domain of the Fc region, and several sites of methionine oxidation. Sequence modifications to the protein and changes in fermentation conditions resulted in increased potency and greater consistency of the product. The final product was shown to be biologically active in animal studies. PMID- 12767821 TI - Expression of caltrin in the baculovirus system and its purification in high yield and purity by cobalt (II) affinity chromatography. AB - Direct protein extraction from animals is the only approach available to obtain caltrin, calcium transport inhibitor. Here we report the expression and purification of caltrin, previously shown to hinder the influx of calcium into epididymal spermatozoa. Cloning of the caltrin gene into the pCDNA3.1 V5/His-TOPO vector and the subsequent ligation of the caltrin-His sequence into the transfer vector pBacPAK9 allowed the expression of recombinant caltrin using the baculovirus expression vector system (BEVS). Recombinant His-tagged caltrin was purified utilising both nickel (II)-nitrilotriacetic acid (Ni(2+)-NTA) and cobalt (II)-carboxymethylaspartate (Co(2+)-CmAsp) immobilised metal affinity chromatography (IMAC). Using the BEVS, caltrin-His was identified in the supernatant and in the cell lysate, suggesting that caltrin is a secreted protein. Based on sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS PAGE) and Western blot results, purified recombinant caltrin-His was ascertained to be approximately 14.5kDa. Purification under the Co(2+) system yielded significantly purer protein samples when compared to the Ni(2+) system. Furthermore, Co(2+) was observed to bind the recombinant caltrin-His protein with higher efficiency and specificity and to yield a higher total protein concentration. Collectively, our results indicate that the Co(2+) system would be a better approach for purifying caltrin-His proteins than the Ni(2+). PMID- 12767823 TI - Purification of glucose 6-phosphate dehydrogenase from Buffalo (Bubalus bubalis) erythrocytes and investigation of some kinetic properties. AB - Glucose 6-phosphate dehydrogenase (G6PD) was purified from buffalo (Bubalus bubalis) erythrocytes and some characteristics of the enzyme were investigated. The purification procedure was composed of two steps: hemolysate preparation and 2('),5(')-ADP-Sepharose 4B affinity gel chromatography. Thanks to the two consecutive procedures, the enzyme, having a specific activity of 69.7EU/mg proteins, was purified 650-fold with a yield of 31%. Optimal pH, stable pH, optimal temperature, molecular weight, and K(M) and V(max) values for NADP(+) and glucose 6-phosphate (G6-P) substrates were also determined for the enzyme. In addition, K(i) values and the type of inhibition were determined by means of Lineweaver-Burk graphs obtained for such inhibitors as ATP, ADP, NADPH, and NADH. PMID- 12767822 TI - Expression, purification, and isotope labeling of the Fv of the human HIV-1 neutralizing antibody 447-52D for NMR studies. AB - The Fv is the smallest antigen binding fragment of the antibody and is made of the variable domains of the light and heavy chains, V(L) and V(H), respectively. The 26-kDa Fv is amenable for structure determination in solution using multi dimensional hetero-nuclear NMR spectroscopy. The human monoclonal antibody 447 52D neutralizes a broad spectrum of HIV-1 isolates. This anti-HIV-1 antibody elicited in an infected patient is directed against the third variable loop (V3) of the envelope glycoprotein (gp120) of the virus. The V3 loop is an immunodominant neutralizing epitope of HIV-1. To obtain the 447-52D Fv for NMR studies, an Escherichia coli bicistronic expression vector for the heterodimeric 447-52D Fv and vectors for single chain Fv and individually expressed V(H) and V(L) were constructed. A pelB signal peptide was linked to the antibody genes to enable secretion of the expressed polypeptides into the periplasm. For easy cloning of any antibody gene without potential modification of the antibody sequence, restriction sites were introduced in the pelB sequence and following the termination codon. A set of oligonucleotides that prime the leader peptide genes of all potential antibody human antibodies were designed as backward primers. The forward primers for the V(L) and V(H) were based on constant region sequences. The 447-52D Fv could not be expressed either by a bicistronic vector or as single chain Fv, probably due to its toxicity to Escherichia coli. High level of expression was obtained by individual expression of the V(H) and the V(L) chains, which were then purified and recombined to generate a soluble and active 447-52D Fv fragment. The V(L) of mAb 447-52D was uniformly labeled with 13C and 15N nuclei (U-13C/15N). Preliminary NMR spectra demonstrate that structure determination of the recombinant 447-52D Fv and its complex with V3 peptides is feasible. PMID- 12767824 TI - A less laborious approach to the high-throughput production of recombinant proteins in Escherichia coli using 2-liter plastic bottles. AB - Contemporary approaches to biology often call for the high-throughput production of large amounts of numerous proteins for structural or functional studies. Even with the highly efficient protein expression systems developed in Escherichia coli, production of these proteins is laborious and time-consuming. We have simplified established protocols by the use of disposable culture vessels: common 2-liter polyethylene terephthalate beverage bottles. The bottles are inexpensive, fit conveniently in commonly available flask holders, and, because they are notched, provide sufficient aeration to support the growth of high-density cultures. The use of antibiotics and freshly prepared media alleviates the need for sterilization of media and significantly reduces the labor involved. Uninoculated controls exhibited no growth during the time required for protein expression in experimental cultures. The yield, solubility, activity, and pattern of crystallization of proteins expressed in bottles were comparable to those obtained under conventional culture conditions. After use, the bottles are discarded, reducing the risk of cross-contamination of subsequent cultures. The approach appears to be suitable for high-throughput production of proteins for structural or functional studies. PMID- 12767825 TI - Crystal structure of a family 45 endoglucanase from Melanocarpus albomyces: mechanistic implications based on the free and cellobiose-bound forms. AB - Cellulose, a polysaccharide of beta-1,4-linked D-glucosyl units, is the major component of plant cell walls and one of the most abundant biopolymers in nature. Cellulases (cellobiohydrolases and endoglucanases) are enzymes that catalyse the hydrolysis of cellulose to smaller oligosaccharides, a process of paramount importance in biotechnology. The thermophilic fungus Melanocarpus albomyces produces a 20 kDa endoglucanase known as 20K-cellulase that has been found particularly useful in the textile industry. The crystal structures of free 20K cellulase and its complex with cellobiose have been determined at 2.0 A resolution. The enzyme, classified into the glycoside hydrolase family 45, exhibits the characteristic six-stranded beta-barrel found before in Humicola insolens endoglucanase V structure. However, the active site in the 20K-cellulase shows a closing of approximately 2.5-3.5A while a mobile loop identified previously in Humicola insolens endoglucanase V and implicated in the catalytic mechanism is well-defined in 20K-cellulase. In addition, the crystal structure of the cellobiose complex shows a shift in the cellobiose position at the substrate binding cleft. It is therefore proposed that these alterations may reflect differences in the binding mechanism and catalytic action of the enzyme. PMID- 12767826 TI - Analysis of NCp7-dependent activation of HIV-1 cDNA integration and its conservation among retroviral nucleocapsid proteins. AB - HIV-1 nucleocapsid protein NCp7 is a small basic protein with two zinc fingers, found in the virion core where several hundred molecules coat the genomic RNA. NCp7 has nucleic acid chaperone properties that guide reverse transcriptase (RT) to synthesize the proviral DNA flanked by the long terminal repeats (LTR). In vitro, NCp7 can strongly activate magnesium-dependent LTR-DNA strand transfer by integrase (IN). Here we show that IN activation relies on both the basic residues and the zinc fingers of NCp7. NCp7 lacking the zinc fingers binds DNA but moderately stimulates strand transfer by IN. The NCp7 zinc-finger domain binds DNA poorly and does not efficiently stimulate IN activity. However, the NC zinc finger domain can complement DNA binding to restore full activation of strand transfer by IN. We propose that the basic residues and the zinc fingers function together to stabilize IN at the LTR ends and promote the formation of a nucleoprotein complex competent for integration. We also show that these properties of HIV-1 NCp7 are remarkably conserved among nucleocapsid proteins of retrotransposon and retrovirus origins. PMID- 12767827 TI - Dynamics and DNA substrate recognition by the catalytic domain of lambda integrase. AB - Bacteriophage lambda integrase (lambda-Int) is the prototypical member of a large family of enzymes that catalyze site-specific DNA recombination via the formation of a Holliday junction intermediate. DNA strand cleavage by lambda-Int is mediated by nucleophilic attack on the scissile phosphate by a conserved tyrosine residue, forming an intermediate with the enzyme covalently attached to the 3' end of the cleaved strand via a phosphotyrosine linkage. The crystal structure of the catalytic domain of lambda-Int (C170) obtained in the absence of DNA revealed the tyrosine nucleophile at the protein's C terminus to be located on a beta hairpin far from the other conserved catalytic residues and adjacent to a disordered loop. This observation suggested that a conformational change in the C terminus of the protein was required to generate the active site in cis, or alternatively, that the active site could be completed in trans by donation of the tyrosine nucleophile from a neighboring molecule in the recombining synapse. We used NMR spectroscopy together with limited proteolysis to examine the dynamics of the lambda-Int catalytic domain in the presence and absence of DNA half-site substrates with the goal of characterizing the expected conformational change. Although the C terminus is indeed flexible in the absence of DNA, we find that conformational changes in the tyrosine-containing beta-hairpin are not coupled to DNA binding. To gain structural insights into C170/DNA complexes, we took advantage of mechanistic conservation with Cre and Flp recombinases to model C170 in half-site and tetrameric Holliday junction complexes. Although the models do not reveal the nature of the conformational change required for cis cleavage, they are consistent with much of the available experimental data and provide new insights into the how trans complementation could be accommodated. PMID- 12767828 TI - Interactions of the Escherichia coli DnaB helicase hexamer with the replication factor the DnaC protein. Effect of nucleotide cofactors and the ssDNA on protein protein interactions and the topology of the complex. AB - Quantitative studies of interactions between the Escherichia coli replication factor DnaC protein and the DnaB helicase have been performed using sedimentation velocity and fluorescence energy transfer techniques. The applied novel analysis of the sedimentation data allows us to construct thermodynamic rigorous binding isotherms without any assumption as to the relationship between the observed molecular property of the complexes formed, the average sedimentation coefficient, or the degree of binding. Experiments have been performed with the fluorescein-modified DnaB helicase, which allows an exclusive monitoring of the DnaB-DnaC complex formation. The DnaC binding to the unmodified helicase has been characterized in competition experiments. The data establish that, in the presence of the ATP analog AMP-PNP, or ADP, a maximum of six DnaC monomers bind cooperatively to the DnaB hexamer. The positive cooperative interactions are limited to the two neighboring DnaC molecules. Analyses using a statistical thermodynamic hexagon model indicate that, under the solution conditions examined, the affinity is characterized by the intrinsic binding constant K=1.4(+/-0.5)x10(5)M(-1) and cooperativity parameter sigma=21+/-5. These data suggest strongly that the DnaC-DnaB complex exists in vivo as a mixture of complexes with a different number of bound DnaC molecules, although the complex with six DnaC molecules bound dominates the distribution. The DnaC nucleotide binding site is not involved in the stabilization of the complex. Moreover, the hydrolysis of NTP bound to the helicase or the DnaC is not required for the release of the DnaC protein from the complex. The single-stranded DNA (ssDNA) bound to the helicase does not affect the DnaC protein binding. However, in the presence of the DNA, there is a significant difference in the energetics and structure of the ternary complex, DnaC-DnaB-ssDNA, formed in the presence of AMP PNP as compared to ADP. The topology of the ternary complex DnaC-DnaB-ssDNA has been determined using the fluorescence energy transfer method. In solution, the DnaC protein-binding site is located on the large 33 kDa domain of the DnaB helicase. The significance of the results in the functioning of the DnaB helicase DnaC protein complex is discussed. PMID- 12767829 TI - The multi-layered structure of Dps with a novel di-nuclear ferroxidase center. AB - The crystallization of cellular components represents a unique survival strategy for bacterial cells under stressed conditions. A highly ordered, layered structure is often formed in such a process, which may involve one or more than one type of bio-macromolecules. The main advantage of biocrystallization has been attributed to the fact that it is a physical process and thus is independent of energy consumption. Dps is a protein that crystallizes to form a multi-layered structure in starved cells in order to protect DNA against oxidative damage and other detrimental factors. The multi-layered crystal structure of a Dps protein from Bacillus brevis has been revealed for the first time at atomic resolution in the absence of DNA. Inspection of the structure provides the first direct evidence for the existence of a di-nuclear ferroxidase center, which possesses unique features among all the di-iron proteins identified so far. It constitutes the structural basis for the ferroxidase activity of Dps in the crystalline state as well as in solution. This finding proves that the enzymatic process of detoxification of metal ions, which may cause severe oxidative damage to DNA, is the other important aspect of the defense mechanism performed by Dps. In the multi-layered structure, Dps dodecamers are organized in a highly ordered manner. They adopt the classic form of hexagonal packing in each layer of the structure. Such arrangement results in reinforced structural features that would facilitate the attraction and absorption of metal ions from the environment. The highly ordered layered structure may provide an ideal basis for the accommodation of DNA between the layers so that it can be isolated and protected from harmful factors under stress conditions. PMID- 12767831 TI - Spontaneous subunit exchange and biochemical evidence for trans autophosphorylation in a dimer of Escherichia coli histidine kinase (EnvZ). AB - The EnvZ/OmpR histidyl-aspartyl phosphorelay (HAP) system in Escherichia coli regulates the expression of ompF and ompC, the major outer membrane porin genes, in response to environmental osmolarity changes. Here, we report that dimers of EnvZc, the cytoplasmic domain of EnvZ, undergo spontaneous subunit exchange in solution. By introducing a cysteine substitution (S260C) in the dimerization domain of EnvZc, we were able to crosslink the two subunits in a dimer and trap the heterodimer formed between two different mutant EnvZc. By using a complementing system with two autophosphorylation-defective EnvZc mutants, one containing the H243V mutation at the autophosphorylation site and the other containing the G405A mutation in the ATP-binding domain, we demonstrated that an EnvZc(G405A) subunit can be phosphorylated by an EnvZc(H243V) subunit only when a heterodimer is formed. The rate of subunit exchange is concentration-dependent, with higher rates at higher concentrations of protein. The disulfide-crosslinked EnvZc(G405A) homodimer could not be phosphorylated by EnvZc(H243V), since the heterodimer formation between the two mutant proteins was blocked, indicating that autophosphorylation cannot occur by dimer-dimer interaction. By using MBP deltaL-EnvZc(S260C) fusion protein (deltaL: the linker region, spanning residues 180-222, was deleted), it was found that in the disulfide-crosslinked MBP-deltaL EnvZc(S260C)/deltaL-EnvZc(S260C/G405A) heterodimer, only the deltaL EnvZc(S260C/G405A) subunit was phosphorylated but not the MBP-deltaL-EnvZc(S260C) subunit. Together, the present results provide biochemical evidence that EnvZ autophosphorylation occurs in trans and only within an EnvZ dimer. PMID- 12767830 TI - Isolation of proteins that interact with the signal transduction molecule Dof and identification of a functional domain conserved between Dof and vertebrate BCAP. AB - Dof is a large molecule essential for signal transduction by the two FGF receptors in Drosophila. It contains two ankyrin repeats and a coiled-coil region, but has no other recognisable structural motif. Dof shares these features with its closest vertebrate relatives, the B-cell signalling molecules BCAP and BANK. In addition, this family of proteins shares a region of homology upstream of the ankyrin repeats, which we call the Dof/BCAP/BANK (DBB) motif. We have identified 44 proteins that interact with Dof in a yeast two-hybrid screen. These include the Drosophila FGF-receptor Heartless and Dof itself. We show that the integrity of the DBB motif is required both for Dof and for BCAP to form dimers. Analysis of the interactions between a set of deletion constructs of Dof and the panel of interactors suggests that Dof may adopt different conformations, with a folded conformation stabilized by interactions between the DBB motif and the C terminal part of the protein. PMID- 12767832 TI - Structural insights into the activation of P. vivax plasmepsin. AB - The malarial aspartic proteinases (plasmepsins) have been discovered in several species of Plasmodium, including all four of the human malarial pathogens. In P.falciparum, plasmepsins I, II, IV and HAP have been directly implicated in hemoglobin degradation during malaria infection, and are now considered targets for anti-malarial drug design. The plasmepsins are produced from inactive zymogens, proplasmepsins, having unusually long N-terminal prosegments of more than 120 amino acids. Structural and biochemical evidence suggests that the conversion process of proplasmepsins to plasmepsins differs substantially from the gastric and plant aspartic proteinases. Instead of blocking substrate access to a pre-formed active site, the prosegment enforces a conformation in which proplasmepsin cannot form a functional active site. We have determined crystal structures of plasmepsin and proplasmepsin from P.vivax. The three-dimensional structure of P.vivax plasmepsin is typical of the monomeric aspartic proteinases, and the structure of P.vivax proplasmepsin is similar to that of P.falciparum proplasmepsin II. A dramatic refolding of the mature N terminus and a large (18 degrees ) reorientation of the N-domain between P.vivax proplasmepsin and plasmepsin results in a severe distortion of the active site region of the zymogen relative to that of the mature enzyme. The present structures confirm that the mode of inactivation observed originally in P.falciparum proplasmepsin II, i.e. an incompletely formed active site, is a true structural feature and likely represents the general mode of inactivation of the related proplasmepsins. PMID- 12767833 TI - The extended multidomain solution structures of the complement protein Crry and its chimeric conjugate Crry-Ig by scattering, analytical ultracentrifugation and constrained modelling: implications for function and therapy. AB - Complement receptor-related gene/protein y (Crry) is a cell membrane-bound regulator of complement activation found in mouse and rat. Crry contains only short complement/consensus repeat (SCR) domains. X-ray and neutron scattering was performed on recombinant rat Crry containing the first five SCR domains (rCrry) and mouse Crry with five SCR domains conjugated to the Fc fragment of mouse IgG1 (mCrry-Ig) in order to determine their solution structures at medium resolution. The radius of gyration R(G) of rCrry was determined to be 4.9-5.0 nm, and the R(G) of the cross-section was 1.2-1.5 nm as determined by X-ray and neutron scattering. The R(G) of mCrry-Ig was 6.6-6.7 nm, and the R(G) of the cross section were 2.3-2.4 nm and 1.3 nm. The maximum dimension of rCrry was 18 nm and that for mCrry-Ig was 26 nm. The neutron data indicated that rCrry and mCrry-Ig have molecular mass values of 45,000 Da and 140,000 Da, respectively, in agreement with their sequences, and sedimentation equilibrium data supported these determinations. Time-derivative velocity experiments gave sedimentation coefficients of 2.4S for rCrry and 5.4S for mCrry-Ig. A medium-resolution model of rCrry was determined using homology models that were constructed for the first five SCR domains of Crry from known crystal and NMR structures, and linked by randomly generated linker peptide conformations. These trial-and-error calculations revealed a small family of extended rCrry structures that best accounted for the scattering and ultracentrifugation data. These were shorter than the most extended rCrry models as the result of minor bends in the inter-SCR orientations. The mCrry-Ig solution data were modelled starting from a fixed structure for rCrry and the crystal structure of mouse IgG1, and was based on conformational searches of the hinge peptide joining the mCrry and Fc fragments. The best-fit models showed that the two mCrry antennae in mCrry-Ig were extended from the Fc fragment. No preferred orientation of the antennae was identified, and this indicated that the accessibility of the antennae for the molecular targets C4b and C3b was not affected by the covalent link to Fc. A structural comparison between Crry and complement receptor type 1 indicated that the domain arrangement of Crry SCR 1-3 is as extended as that of the CR1 SCR 15-17 NMR structure. PMID- 12767834 TI - The effects of mutations on motions of side-chains in protein L studied by 2H NMR dynamics and scalar couplings. AB - Recently developed 2H spin relaxation experiments are applied to study the dynamics of methyl-containing side-chains in the B1 domain of protein L and in a pair of point mutants of the domain, F22L and A20V. X-ray and NMR studies of the three variants of protein L studied here establish that their structures are very similar, despite the fact that the F22L mutant is 3.2kcal/mol less stable. Measurements of methyl 2H spin relaxation rates, which probe dynamics on a picosecond-nanosecond time scale, and three-bond 3J(Cgamma-CO), 3J(Cgamma-N) and 3J(Calpha-Cdelta) scalar coupling constants, which are sensitive to motion spanning a wide range of time-scales, reveal changes in the magnitude of side chain dynamics in response to mutation. Observed differences in the time-scale of motions between the variants have been related to changes in energetic barriers. Of interest, several of the residues with different motional properties across the variants are far from the site of mutation, suggesting the presence of long range interactions within the protein that can be probed through studies of dynamics. PMID- 12767835 TI - The sequence dependence of fiber organization. A comparative molecular dynamics study of the islet amyloid polypeptide segments 22-27 and 22-29. AB - Amyloid fiber formation and the possible polymorphism of molecular arrangements depend on the polypeptide length and composition. Here, we seek the chemical clues underlying these processes. Our starting point is based on the experimental observation that some short peptide segments are able to develop fibers that are very similar to those of their original parent proteins. We focus our study on the NFGAILSS peptide, derived from the human islet amyloid polypeptide (residues 22-29). This peptide turned out to be a perfect example, illustrating the fact that the amyloid microscopic organization is highly complex, rather than simply involving hydrogen bond formation. Furthermore, obtaining a reliable molecular model has allowed us to analyze the differences between the amyloid structure we have obtained for this peptide and that obtained for the previously studied, two residues shorter, segment (residues 22-27, NFGAIL). This comparative study yields some clues about chemical events that govern the aggregation of proteins into oriented fibers, such as molecular packing between sheets and the degree of interaction specificity. We characterize the important role played by the hydrophobic and aromatic residues in the inter-sheet association and present new approaches toward the understanding of the nature of events that are likely to take place during fibril formation. These include analysis of interaction patterns derived from specific sheet-associated packing. PMID- 12767836 TI - Folding rates and low-entropy-loss routes of two-state proteins. AB - We develop a simple model for computing the rates and routes of folding of two state proteins from the contact maps of their native structures. The model is based on the graph-theoretical concept of effective contact order (ECO). The model predicts that proteins fold by "zipping up" in a sequence of small-loop closure events, depending on the native chain fold. Using a simple equation, with a few physical rate parameters, we obtain a good correlation with the folding rates of 24 two-state folding proteins. The model rationalizes data from Phi value analysis that have been interpreted in terms of delocalized or polarized transition states. This model indicates how much of protein folding may take place in parallel, not along a single reaction coordinate or with a single transition state. PMID- 12767837 TI - The folding mechanism of a two-domain protein: folding kinetics and domain docking of the gene-3 protein of phage fd. AB - The gene-3 protein (G3P) of filamentous phages is essential for the infection of Escherichia coli. The carboxy-terminal domain anchors this protein in the phage coat, whereas the two amino-terminal domains N1 and N2 protrude from the phage surface. We analyzed the folding mechanism of the two-domain fragment N1-N2 of G3P (G3P(*)) and the interplay between folding and domain assembly. For this analysis, a variant of G3P(*) was used that contained four stabilizing mutations (IIHY-G3P(*)). The observed refolding kinetics extend from 10 ms to several hours. Domain N1 refolds very rapidly (with a time constant of 9.4 ms at 0.5 M guanidinium chloride, 25 degrees C) both as a part of IIHY-G3P(*) and as an isolated protein fragment. The refolding of domain N2 is slower and involves two reactions with time constants of seven seconds and 42 seconds. These folding reactions of the individual domains are followed by a very slow, spectroscopically silent docking process, which shows a time constant of 6200 seconds. This reaction was detected by a kinetic unfolding assay for native molecules. Before docking, N1 and N2 unfold fast and independently, after docking they unfold slowly in a correlated fashion. A high energy barrier is thus created by domain docking, which protects G3P kinetically against unfolding. The slow domain docking is possibly important for the infection of E.coli by the phage. Upon binding to the F pilus, the N2 domain separates from N1 and the binding site for TolA on domain N1 is exposed. Since domain reassembly is so slow, this binding site remains accessible until pilus retraction has brought N1 close to TolA on the bacterial surface. PMID- 12767838 TI - Prudent modeling of core polar residues in computational protein design. AB - Hydrogen bond interactions were surveyed in a set of protein structures. Compared to surface positions, polar side-chains at core positions form a greater number of intra-molecular hydrogen bonds. Furthermore, the majority of polar side-chains at core positions form at least one hydrogen bond to main-chain atoms that are not involved in hydrogen bonds to other main-chain atoms. Based on this structural survey, hydrogen bond rules were generated for each polar amino acid for use in protein core design. In the context of protein core design, these prudent polar rules were used to eliminate from consideration polar amino acid rotamers that do not form a minimum number of hydrogen bonds. As an initial test, the core of Escherichia coli thioredoxin was selected as a design target. For this target, the prudent polar strategy resulted in a minor increase in computational complexity compared to a strategy that did not allow polar residues. Dead-end elimination was used to identify global minimum energy conformations for the prudent polar and no polar strategies. The prudent polar strategy identified a protein sequence that was thermodynamically stabilized by 2.5 kcal/mol relative to wild-type thioredoxin and 2.2 kcal/mol relative to a thioredoxin variant whose core was designed without polar residues. PMID- 12767840 TI - Lack of influence of body exhaust gowns on aerobic bacterial surface counts in a mixed-ventilation operating theatre. A study of 62 hip arthroplasties. AB - Aerobic bacterial surface contamination was studied with and without the use of body exhaust gowns in an operating room equipped with mixed/turbulent ventilation and separate operating and anaesthetic areas during 62 hip joint arthroplasties. In 31 operations conventional gowns were used, and 31 were performed with body exhaust gowns. Bacterial surface contamination was monitored in the operating and anaesthetic area using 9 cm diameter settle plates (1+1) and nitrocellulose membranes (2+2) transferred after sampling to nutrient pads. Compared with conventional clothing, the use of body exhaust gowns did not significantly reduce the microbial contamination (P=0.1-0.7). On the settle plates 1 m from the patient 279+/-326 cfu/m(2)/h were observed with conventional clothing compared with 142+/-227 cfu/m(2)/h with body exhaust gowns. The first membrane located on the patient in the sterile area detected 250+/-590 cfu/m(2)/h with conventional clothing and 210+/-320 cfu/m(2)/h with exhaust gowns. For the second membrane on the floor, the counts were 1790+/-2700 and 1590+/-1590 cfu/m(2)/h. For all operations the settle plates yielded 210+/-287 cfu/m(2)/h in the operating area and 720+/-564 cfu/m(2)/h in the anaesthetic area (P=0.01). Compared with the membrane placed on the anaesthetic equipment the counts on the membrane placed on patient were also significantly lower (P=0.01) while the membranes placed on the floor in each area showed no difference in counts. In conclusion, compared with conventional clothing, the use of body exhaust gowns could not be proven to provide more protection against microbial contamination. The low number of colony forming units found in the operating area was similar to that expected from an ultraclean laminar airflow unit, although achieved with a cheaper and more energy saving system. PMID- 12767841 TI - Effect of central venous catheter type on infections: a prospective clinical trial. AB - This study reports on a block clinical trial of two types of central venous catheters (CVCS): antiseptic-impregnated catheters (AIC) and non-impregnated catheters (non-AIC), on catheter tip colonization and bacteraemia. In total, 500 catheters were inserted in 390 patients over the 18 month study period, 260 (52.0%) AIC and 240 (48.0%) non-AIC. Of these, 460 (92.0%) tips (237 AIC and 223 non-AIC) were collected. While significantly fewer AIC, 14 (5.9%), than non-AIC, 30 (13.5%), catheters were colonized (P<0.01), there was no difference in the rates of bacteraemias in the two groups (0.8% vs. 2.7%, respectively, P=0.16). There were 6.87 (95% CI 3.38-14.26) and 16.92 (95% CI 10.61-27.12) colonized AIC and non-AIC catheters, respectively, per 1000 catheter days, a difference that was significant (P<0.01). However, no difference emerged between bacteraemias in AIC and non-AIC catheters per 1000 catheter days measured at 0.98 (95% CI 0.24 5.54) and 3.38 (95% CI 1.29-9.34), respectively (P=0.10). Of the 444 CVCs that were sited in the subclavian or jugular veins and had tips collected, significantly more catheters were colonized in the jugular group, 19 (20%), compared with the subclavian group, 24 (6.9%; P< or =0.01). Overall, the low rates of colonization and bacteraemia may be explained by the population studied, the policies used and the employment of a clinical nurse dedicated to CVC management. PMID- 12767842 TI - Management and outcome of bloodstream infections due to Candida species in England and Wales. AB - This two-year prospective hospital population-based study of candidaemia is the first to be conducted in the UK. It was carried out on behalf on the British Society for Medical Mycology (BSMM) as part of the European Confederation of Medical Mycology (ECMM) epidemiological survey of candidaemia. Six hospitals in England and Wales acted as sentinel hospitals. Main outcome measures were hospital population-based incidence and 30-day mortality. There were 18.7 episodes of candidaemia per 100,000 finished consultant episodes or 3.0/100,000 bed days and 45.4% cases occurred in intensive care unit (ICU) patients. Candida albicans was isolated in 64.7% of confirmed cases. The majority of isolates were sensitive to standard antifungal agents, including fluconazole. The overall 30 day mortality was 26.4% and removal of the central venous catheter was associated with a significant reduction in mortality. In conclusion, the incidence of candidaemia in England and Wales is similar to that of the USA, the majority of isolates remain sensitive to commonly used antifungal agents and mortality associated with this infection appears to be falling. PMID- 12767843 TI - A seven-year survey of Klebsiella pneumoniae producing TEM-24 extended-spectrum beta-lactamase in Nice University Hospital (1994-2000). AB - The aim of this study was to investigate TEM-24-producing isolates of Klebsiella pneumoniae, and their clonal dissemination in Nice University Hospital. During the 1994-2000 period, a total of 263 non-repetitive isolates of ESBL-producing K. pneumoniae were collected. Most of these isolates were highly resistant in vitro to ceftazidime, cefotaxime and aztreonam, but susceptible to cefoxitin and imipenem. Resistance profile analysis revealed seven predominant antibiotypes (P1 to P7). Isoelectric focusing evidenced beta-lactamase activity, with a chromosomal penicillinase (pl 7.7), and one or two additional enzymes with pls ranging from 5.4 to 8.2 identified as presumed TEM-1 pl 5.4, TEM-3 pl 6.3, TEM-24 pl 6.5, SHV-3 pl 7.0, SHV-4 pl 7.8, SHV-5 pl 8.2, or other unidentified beta lactamases. Among these K. pneumoniae, 130 isolates produced TEM-24, and 115 of them were highly resistant in vitro to quinolones (antibiotype P1). This phenotype was responsible for an outbreak in a medical intensive care unit from March to September 2000. Four isolates submitted were genetical sequenced, and shared 99.9% homology with tem-24 (GenBank no. X 65253). Pulsed-field gel electrophoresis and enterobacterial repetitive intergenic consensus polymerase chain reaction (PCR) (ERIC2-PCR) applied to 28 non-epidemic and six epidemic isolates yielded concordant results. Molecular typing revealed the persistence and dissemination of a single clone of TEM-24 producing K. pneumoniae in Nice Hospital during the seven-year study period. PMID- 12767845 TI - Bacteriological, clinical and epidemiological characteristics of hospital acquired Acinetobacter baumannii infection in a teaching hospital. AB - Over an 18 month period, the bacteriological, clinical and epidemiological characteristics of nosocomial Acinetobacter baumannii infections in a teaching hospital were studied. Typing studies were performed on 38 strains isolated from 36 patients. Twenty-two of the strains were isolated during the three outbreaks. Surgery, catheterization, mechanical ventilation, and antibiotic therapy for adult patients and respiratory distress syndrome, mechanical ventilation, and prematurity for paediatric patients were the main risk factors identified. All isolates were resistant to penicillins (except ampicillin-sulbactam), cephalosporins, gentamicin, and aztreonam but susceptible to carbapenems and colistin. Resistance to tobramycin, ciprofloxacin, ampicillin-sulbactam, trimethoprim-sulfamethoxazole, and amikacin was variable. Antibiotyping, arbitrarily-primed polymerase chain reaction (AP-PCR) and the pulse-field gel electrophoresis (PFGE) indicated the epidemiological relationship. The outbreak strains, demonstrated genetic distinction between our three outbreaks and isolates from specific areas in the hospital. PMID- 12767844 TI - Nosocomial multi-drug resistant Acinetobacter baumannii bloodstream infection: risk factors and outcome with ampicillin-sulbactam treatment. AB - The emergence of multidrug-resistant (MDR) Acinetobacter baumannii poses a therapeutic problem. The aim of this study was to assess the risk factors for nosocomial MDR-A. baumannii bloodstream infection (BSI) and the efficacy of ampicillin-sulbactam (A/S) in its treatment. Of 94 nosocomial A. baumannii BSI during the year 2000, 54% involved MDR strains, 81% of which were genetically related. Various risk factors for MDR-A. baumannii were found, of which intensive care unit admission and prior aminoglycoside therapy were independently associated with MDR-A. baumannii acquisition on multivariate analysis. Of MDR-A. baumannii BSI cases, 65% received A/S and 35% inadequate antibiotic therapy, whereas of 43 non-MDR cases, 86% were treated according to susceptibility and 14% inappropriately with antibiotics to which these organisms were resistant. Crude mortality was comparable in the adequately treated groups. Respective mortalities among patients treated adequately and inadequately were 41.4 and 91.7% (p<0.001). Among severely ill patients, A/S therapy significantly decreased the risk of death (P=0.02 OR=7.64). MDR-A. baumannii has become highly endemic in our institution. A/S appears to be one of the last effective and safe empirical resorts for treatment of MDR A. baumannii BSI. PMID- 12767846 TI - In-use evaluation of Perasafe compared with Cidex in fibreoptic bronchoscope disinfection. AB - The mycobactericidal activity of Perasafe (0.26% peracetic acid) was compared with that of Cidex (2% alkaline glutaraldehyde) by an in-use test. Fibreoptic bronchoscopes were artificially contaminated with Mycobacterium tuberculosis or Mycobacterium avium-intracellulare in sputum and, after manual pre-cleaning with a neutral soap, 10 and 20 min disinfection periods were tested. Perasafe was as effective as Cidex, thus requiring a 10 min disinfection period against M. tuberculosis and 20 min against M. avium-intracellulare. The results demonstrate that Perasafe is an effective disinfectant for use in reprocessing fibreoptic bronchoscopes. PMID- 12767847 TI - In-vitro evaluation of Perasafe compared with 2% alkaline glutaraldehyde against Mycobacterium spp. AB - Quantitative suspension and carrier tests were used to compare the activity of Perasafe and Cidex against Mycobacterium tuberculosis, Mycobacterium avium intracellulare, Mycobacterium fortuitum, and Mycobacterium chelonae. The interference of an organic load, and of hard water was also considered. Both agents achieved reductions exceeding 10(5)-fold within 20 and 30 min for all the strains tested. Perasafe is thus mycobactericidal and a viable alternative to Cidex for intermediate or high-level disinfection. PMID- 12767848 TI - Modelling the usefulness of a dedicated cohort facility to prevent the dissemination of MRSA. AB - The aim of this retrospective study was to determine whether or not a surgical dedicated cohort facility, mainly dedicated to the care of orthopaedic patients, can control the risk of infection caused by methicillin-resistant Staphylococcus aureus (MRSA). We tested this hypothesis on the orthopaedic surgery ward of a university-affiliated public hospital with 1228 beds by determining whether there was a significant correlation between the colonization pressure exerted by MRSA and the number of cases of acquired MRSA. This was then used as a tool to predict the number of patients contaminated with MRSA in hospitals with and without dedicated cohort facilities. We found that the relative risk of MRSA acquisition increased with the colonization pressure exerted by MRSA imported cases. This statistical model enabled us to predict that the risk of MRSA acquisition would increase by 160% per year in the absence of a dedicated cohort facility. We conclude that these units are useful to control the spread of MRSA in hospitals. PMID- 12767849 TI - Evaluation of the bactericidal effect of five products for surgical hand disinfection according to prEN 12054 and prEN 12791. AB - Surgical hand disinfection (with an alcohol-based hand rub) and surgical handwash (with an antiseptic-based liquid soap) are accepted measures to reduce the risk for surgical site infections. The new European Standards allow a comparison of their antimicrobial efficacy. The bactericidal activity of surgical hand rubs [Sterillium and Softaman, (active ingredient=alcohols)] and handwashes [Derman plus (triclosan), Hibiscrub (chlorhexidine) and Betadine (PVP-iodine)] was tested according to the prEN 12054 suspension test using Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Enterococcus hirae, and to prEN 12791 for the effect on resident skin flora in comparison with 1-propanol, 60% (v/v). All five products achieved a reduction of test bacteria within 3 min of >10(5)-fold so fulfilling prEN 12054. However, only Hibiscrub, Sterillium and Softa Man met the requirements of prEN 12791, giving a mean reduction of resident micro organisms (immediate and sustained effect) which was not significantly lower than the reference alcohol (P>0.1; Wilcoxon matched-pairs signed-rank test). Sterillium was significantly more effective than the reference alcohol (immediate and sustained affect). Products for surgical hand disinfection may have equal antimicrobial activity in suspension tests but show large differences under practical conditions. Healthcare workers should not rely on results from suspension tests when deciding on a product for surgical hand disinfection. PMID- 12767850 TI - A survey of doctors' and nurses' knowledge, attitudes and compliance with infection control guidelines in Birmingham teaching hospitals. AB - This study investigated knowledge about infection control amongst doctors and nurses through a cross-sectional survey conducted between March and May 2001 in three Birmingham, UK teaching hospitals. Seventy-five doctors and 143 nurses, representing 7% and 4%, respectively, of potential respondents, participated in the study measuring knowledge of, attitudes towards, and compliance with universal precautions. Overall knowledge of risks of blood-borne virus (BBV) transmission from an infected patient after needlestick injury was low [44.0% for hepatitis B virus (HBV), 38.1% for hepatitis C virus (HCV), 54.6% for human immunodeficiency virus (HIV)]. There were significant differences between doctors and nurses concerning the estimations of HBV (e-antigen +) (P=0.006) and HIV (P<0.001) transmission risks. Eighty-six percent of nurses stated that they treat each patient as if they are carrying a BBV compared with 41% of doctors. Doctors and nurses differed significantly in their attitudes about and reported compliance with washing hands before and after patient contact and with wearing gloves when taking blood (P<0.001 for all). Doctors consistently de-emphasized the importance of, and reported poor compliance with, these procedures. Doctors were also more likely to state that they re-sheath used needles manually than were nurses (P<0.001). Thirty-seven percent of respondents reported that they had suffered a needlestick injury with a used needle, with doctors more likely to be injured than nurses (P=0.005). Twenty-eight percent of these doctors and 2% of the nurses did not report their needlestick injuries (P=0.004). Education, monitoring, improved availability of resources, and disciplinary measures for poor compliance are necessary to improve infection control in hospitals, especially amongst doctors. PMID- 12767851 TI - Evidence to support the existence of subgroups within the UK epidemic Clostridium difficile strain (PCR ribotype 1). AB - We used three different DNA fingerprinting techniques and clindamycin susceptibility testing to confirm that Clostridium difficile PCR ribotype I isolates can be divided into two subclones. This observation may permit a better understanding of the epidemiology and pathogenicity of C. difficile infection. PMID- 12767852 TI - Static electric charge may contribute to infections in bone marrow transplant wards. PMID- 12767853 TI - The dangers of wall-mounted hand gel dispensers. PMID- 12767854 TI - Endoscopic decontamination: an audit and practical review. PMID- 12767856 TI - Re: Mereghetti, L., Tayoro, J., Watt, S., et al., Genetic relationship between Escherichia coli strains isolated from the intestinal flora and those responsible for infectious diseases among patients hospitalized in intensive care units. J Hosp Infect, 52 (2002) 43-51. PMID- 12767858 TI - Environmental benefits and economic costs of manure incorporation on dairy waste application fields. AB - Model simulations performed representing dairies in a 93000 ha watershed in north central Texas suggest that manure incorporation results in reduced phosphorus (P) losses at relatively small to moderate cost to producers. Simulated manure incorporation with a tandem disk on fields double-cropped with sorghum/winter wheat resulted in up to 33, 45, and 37% reductions in per hectare sediment-bound, soluble, and total P losses in edge-of-field runoff, relative to simulated surface manure applications. The effects of incorporation were evaluated at three different manure application rates. On aggregate across all three manure application rates, significant declines in P losses were obtained with incorporation except for sediment-bound P losses under the N-based manure application rate scenario. We found that the practice of incorporating manure shortly after it has been broadcast on the soil surface could help reduce P losses in such situations where P-based rates alone prove inadequate. The cost the producer incurs when manure is incorporated is on average about 1% of net returns when manure is applied at the N rate and 2-3% when it is applied at alternative P-based rates. In practice the costs could be lower because producers may substitute the manure incorporation operation for a tandem disk operation performed prior to manure application. As more and more dairy producers switch to the use of sorghum and corn silage in dairy rations and consequent on-farm production of these forages, the practice of manure incorporation may help to reduce phosphorus losses resulting from dairy manure applications to fields with these forage crops. PMID- 12767859 TI - Dealing with electronic waste: modeling the costs and environmental benefits of computer monitor disposal. AB - The importance of information technology to the world economy has brought about a surge in demand for electronic equipment. With rapid technological change, a growing fraction of the increasing stock of many types of electronics becomes obsolete each year. We model the costs and benefits of policies to manage 'e waste' by focusing on a large component of the electronic waste stream-computer monitors-and the environmental concerns associated with disposal of the lead embodied in cathode ray tubes (CRTs) used in most monitors. We find that the benefits of avoiding health effects associated with CRT disposal appear far outweighed by the costs for a wide range of policies. For the stock of monitors disposed of in the United States in 1998, we find that policies restricting or banning some popular disposal options would increase disposal costs from about US dollar 1 per monitor to between US dollars 3 and US dollars 20 per monitor. Policies to promote a modest amount of recycling of monitor parts, including lead, can be less expensive. In all cases, however, the costs of the policies exceed the value of the avoided health effects of CRT disposal. PMID- 12767860 TI - Economic growth, biodiversity loss and conservation effort. AB - This paper investigates the relationship between economic growth, biodiversity loss and efforts to conserve biodiversity using a combination of panel and cross section data. If economic growth is a cause of biodiversity loss through habitat transformation and other means, then we would expect an inverse relationship. But if higher levels of income are associated with increasing real demand for biodiversity conservation, then investment to protect remaining diversity should grow and the rate of biodiversity loss should slow with growth. Initially, economic growth and biodiversity loss are examined within the framework of the environmental Kuznets hypothesis. Biodiversity is represented by predicted species richness, generated for tropical terrestrial biodiversity using a species area relationship. The environmental Kuznets hypothesis is investigated with reference to comparison of fixed and random effects models to allow the relationship to vary for each country. It is concluded that an environmental Kuznets curve between income and rates of loss of habitat and species does not exist in this case. The role of conservation effort in addressing environmental problems is examined through state protection of land and the regulation of trade in endangered species, two important means of biodiversity conservation. This analysis shows that the extent of government environmental policy increases with economic development. We argue that, although the data are problematic, the implications of these models is that conservation effort can only ever result in a partial deceleration of biodiversity decline partly because protected areas serve multiple functions and are not necessarily designated to protect biodiversity. Nevertheless institutional and policy response components of the income biodiversity relationship are important but are not well captured through cross-country regression analysis. PMID- 12767861 TI - Modelling the impacts of strategic tree plantings on salt loads and flows in the Macquarie river catchment, NSW, Australia. AB - In Australia, problems of dryland and stream salinity have recently become the focus of a National Action Plan. In many river catchments, preliminary stream salt load and salinity targets have been set to define maximum permissible export levels in 2015. Afforestation has been proposed as a strategy for meeting these targets, although several studies suggest that widespread commercial tree plantations are likely to deliver net dis-benefits. However, the impacts on stream salt loads of more localised tree plantings in high salt yielding areas have not been quantified. In this paper we use a simple empirical model to predict the effects of various strategic and non-strategic tree planting scenarios on flows and salt loads in the mid-Macquarie catchment, New South Wales. A simple salt routing model is then used to estimate the effect of these changes on salt loads at the end-of-valley monitoring site for the Macquarie catchment. Results suggest that widespread land management interventions will be required to meet the preliminary salt load targets for this catchment. On their own, small-scale, strategic tree planting in high salt export areas of the mid Macquarie area will not have a significant impact on salt loads at the end-of valley monitoring site. While widespread tree plantings may reduce salt loads in the longer term, they are likely to cause streamflow losses in the shorter term. Thus, stream salinities are expected to rise initially, due to the different response times of groundwater and surface water systems to land use change. PMID- 12767862 TI - Diversion of water flow from a floodplain wetland stream: an analysis of geomorphological setting and hydrological and ecological consequences. AB - Diversion of water has been ongoing in the Mkuze Wetland for several decades. Two canals form the focus of this study; the Mpempe-Demazane Canal and the Tshanetshe Canal. The former involved an ambitious excavation over a distance of 13.5 km in the lower part of the wetland, while the latter was a minor excavation over a distance of approximately 100 m in the upper part of the wetland. Although ambitious and costly, the Mpempe-Demazane Canal resulted in little downward or headward erosion, and there was minor diversion of flow. However, the minor excavation of the Tshanetshe Canal resulted in erosion downstream of the excavation (the Tshanetshe Stream), downward and lateral erosion of the excavated section, and headward erosion that has propagated almost 4 km upstream along the Mkuze River. Most of the flow of the Mkuze River has been captured by the Tshanetshe Canal and Stream. The impact of canalisation on floodplain wetlands is thus more dependent on the location than the scale of activity. The avulsion of the Mkuze River into the Tshanetshe Canal and Stream is due to a large difference in elevation between the Mkuze River and floodplain into which it was diverted, and the fact that in this region the river typically has high discharges. This avulsion may have been inevitable as a result of natural processes of sedimentation. In contrast, the difference in elevation between the Mkuze River and the basin into which it was diverted via the Mpempe Canal was small as is discharge of the Mkuze River in this part of the wetland. Thus, the diversion was unsuccessful. The presence of hippos that create hydraulically efficient pathways that are oriented parallel to the regional hydraulic slope, may accelerate avulsion in large African wetlands. Overall, it is argued that the environmental consequences of excavation need to be viewed against the background that wetlands are dynamic features within the landscape. PMID- 12767863 TI - The effect of heating technologies on CO(2) and energy efficiency of Dutch greenhouse firms. AB - This paper uses Data Envelopment Analysis to compute measures of the efficiency (relative to a frontier) in terms of the use of all inputs as well as for single inputs like CO(2) and energy for a sample of greenhouse firms in the Netherlands over the period 1991-1995. These efficiency measures are generated for different firms specialised in production of vegetables, flowers, and potplants and with different heating technologies. The empirical results indicate that firms use energy quite efficiently and are less efficient in terms of CO(2) emissions. Firms using conventional heating are overall less efficiently using energy and CO(2) than firms using more advanced heating technologies. Most differences in efficiency between firm types and firms using different heating technologies are statistically significant. Scale adjustments can provide an important contribution to further efficiency improvements. PMID- 12767864 TI - Measuring principals' values for environmental budget management: an exploratory study. PMID- 12767865 TI - Adjusting irrigation abstraction to minimise the impact on stream flow in the East of Scotland. AB - Abstractions of surface and groundwater for irrigation in Scotland are currently subject to control in only two small catchments. Under the terms of the EU Water Framework Directive, it will be necessary to introduce new legislation to control abstractions elsewhere. To help in the development of appropriate policy for Scotland a study has been carried out to examine the significance of irrigation and the effectiveness of different types of control strategies in terms of the economics of potato cropping and stream hydrology in Scotland. This paper presents the findings of the hydrological study and highlights some of the spatial and temporal issues that need to be considered in the selection of control mechanisms, if they are to be successful in achieving objectives for environmental improvement. The study was focussed on two catchments in the east of Scotland, the Tyne and West Peffer. The effectiveness of several different abstraction control strategies was examined to see how stream flows in the catchment would be modified by their implementation. The results of the study demonstrated that the West Peffer catchment in particular is significantly affected by irrigation abstractions. Control mechanisms based on allowable monthly abstraction volumes and flow-based abstraction bans would be of considerable help in restoring stream flows to their natural levels, but would modify the hydrological regime in slightly different ways. A spatial analysis of stream flows demonstrated that implementation of controls based on a single monitoring point may be ineffective at maintaining acceptable levels of flow throughout the catchment and that this may require a tighter control at the monitoring point. PMID- 12767868 TI - Prognostic relevance of molecular markers of oral cancer--a review. AB - The aim of the present article was to review the current knowledge on the prognostic value of tumour marker in the treatment of oral squamous cell carcinoma (OSCC). The literature of the past 5 years (1997-August 2002) was screened. One hundred and sixty-nine articles were included in this review, twenty-nine molecular markers of relevance were identified. Tumour markers were allocated to four groups according to their function: (i) Enhancement of Tumour Growth: Cell cycle acceleration and proliferation, (ii) Tumour Suppression and Anti-Tumour Defence: Immune response and apoptosis, (iii) Angiogenesis, (iv) Tumour Invasion and Metastatic Potential: Adhesion molecules and matrix degradation. Data showed that the prognostic relevance of most tumour markers is still not quite clear. Only 12 of 23 reports on the prognostic relevance of markers for cell cycle acceleration and proliferation indicated a significant association with prognosis while 20 of 29 studies on markers for tumour suppression and anti-tumour response showed prognostic relevance. Markers of angiogenesis exhibited only minor importance for the prognosis and treatment of OSCC. Results on markers of tumour invasion and metastatic potential appeared to be too premature for a statement regarding their prognostic value. In general, the location of markers within the tumour and not their quantitative assessment as such is emphasized. Particularly, the analysis of the invasive front of the tumour with regard to the occurrence of molecular markers is supposed to be of great importance for prognostication. PMID- 12767869 TI - Comparison between direct clinical and digital photogrammetric measurements in patients with 22q11 microdeletion. AB - Thanks to recent advances in imaging and computing technology, photoanthropometry has become an increasingly helpful adjunct to obtain objective clinical evidence of morphologic abnormalities in patients with dysmorphic syndromes. The aim of this study was to evaluate measurements made using a new two-dimensional digital photogrammetry technique by comparison with direct clinical measurements. A total of 14 patients with 22q11 microdeletion were included in this study. There were seven females and seven males between 5 and 38 years of age. Sixteen direct clinical measurements were performed using a graduated anthropometric sliding caliper and angle meter. Photogrammetric measurements were made on digital photographs using a commercially available software package. After calibration to one in situ reference on frontal and profile views, photogrammetric measurements were compared to a total of 14 direct clinical measurements made in the same patients. Findings showed that photogrammetric measurements calibrated to an in situ reference were reliable especially on profile views. No statistical difference was found between 10 of the 14 measurements (P> 0.05) including eight of the nine measurements on profile views and two of the five measurements on frontal views. This study indicates that digital photogrammetry is a useful tool but there is still no reliable standard photographic measurement technique. PMID- 12767870 TI - Retromandibular approach to the mandibular condyle: a clinical and cadaveric study. AB - This prospective study was carried out to assess the morbidity of the retromandibular approach in the management of condylar fractures. Twenty consecutive patients underwent open reduction and internal fixation of their condylar fractures using this technique. Branches of the facial nerve were encountered in six cases (30%). Temporary weakness of the facial nerve occurred in six patients (30%), but this resolved in all cases within 3 months and there were no cases of permanent nerve injury. Two patients had a temporary deficit of the great auricular nerve and one patient developed a sialocoele that resolved with aspiration. A cadaveric study using 30 facial halves (15 fresh cadavers) was also conducted. Branches of the facial nerve were encountered in 12 dissections (40%). The literature regarding facial nerve morbidity in relation to the management of condylar fractures is reviewed. PMID- 12767871 TI - Follow-up study of treatment of orbital floor fractures: relation of clinical data and software-based CT-analysis. AB - This retrospective study quantifies isolated orbital floor fractures using software-based CT-analysis and compares the clinical outcome across surgical and non-surgical treatment groups. Depending on the surgeon's interpretation of the clinical and radiological appearance, 10 fractures were treated non-surgically and 20 fractures surgically, either with antral balloon catheter alone or in combination with an orbital implant. Ophthalmologic findings were evaluated until 12 weeks after injury. Fracture area, and volume of displaced tissue (VDT) were assessed by software-based CT-analysis. VDT was marginally significantly smaller in non-surgically than in surgically-treated patients (P=0.08). Ophthalmologic findings improved in all groups during follow-up and no statistical difference was found between the groups. Diplopia remained moderate in three patients with balloon catheter alone, and minimal in four patients in both surgical groups. In one patient with non-surgical treatment, diplopia remained minimal after 12 weeks. Although CT-analysis revealed no significant difference between both surgical groups, patients treated with balloon catheter alone presented more diplopia after 12 weeks. Using balloon catheters for fracture repair a combined approach should be performed when large fractures involve the orbital floor to achieve sufficient reduction of orbital content and placement of an orbital implant. Software-based CT-analysis is helpful for objective interpretation in managing of orbital fractures. PMID- 12767872 TI - Five-year retrospective evaluation of temporomandibular joint arthrocentesis. AB - This retrospective study is aimed to evaluate the long-term outcome of arthrocentesis for the treatment of temporomandibular joint (TMJ) internal derangements, and to document the impact of patient, time and symptom-related factors on the outcome of the procedure. Thirty-four patients (48 joints) who underwent TMJ arthrocentesis were included in this study. The scores for preoperative maximal mouth opening, and VAS scores for pain and dysfunction were compared with the follow-up scores obtained by the questionnaire and clinical examination. Several factors that may affect the long-term outcome of arthrocentesis are further evaluated. Statistical evaluation of the baseline and follow-up data was made by linear regression analysis and paired t-test. The mean follow-up period was 22 months. There was a significant (P< 0.001) increase in the maximal mouth opening (MMO) postoperatively that held during the longer term follow-up period. The pain and dysfunction levels at the follow-up were significantly (P< 0.001) lower than the preoperative values. Twenty-six per cent of patients were pain free and also showed a total relief in dysfunction at the follow-up. Although both preoperative and follow-up pain scores were higher in patients with bruxism, there was not a significant difference in the outcome when compared with non-bruxers. However, there was a greater reduction in dysfunction with improvement in MMO in non-bruxers. The duration of symptoms before arthrocentesis has not been found to affect the outcome. Also, there were no significant differences between the results of follow-up when comparing the shorter follow-up time results (<20 months) and longer term results. Arthrocentesis for the treatment of TMJ internal derangements offers favourable long-term stable results with regard to increasing maximal mouth opening, and reducing pain and dysfunction. PMID- 12767873 TI - TMJ Concepts/Techmedica custom-made TMJ total joint prosthesis: 5-year follow-up study. AB - This prospective study evaluated the 5 to 8 year subjective and objective results of 42 consecutive patients who had TMJ reconstruction using the TMJ Concepts/Techmedica custom made total joint prosthesis. Criteria for use of the prosthesis included the following TMJ conditions: (1) multiply operated, (2) previous alloplastic implants, (3) osteoarthritis, (4) inflammatory or resorptive arthritis, (5) connective tissue or autoimmune disease, (6) ankylosis, and (7) absent or deformed structures. Thirty-eight of 42 patients (90%) with 69 TMJs reconstructed using the TMJ Concepts/Techmedica total joint prosthesis had appropriate data for inclusion in the study. The average age at surgery was 36 years and average follow-up was 73.5 months. The entire group and three subgroups were objectively evaluated for incisal opening, lateral excursions, and occlusal stability, while subjectively assessed for pain and jaw function. Paired t-test and comparison analyses were used to assess outcomes. For the group of 38 patients, there was statistically significant improvement in incisal opening (P=0.001), jaw function (P=0.001), and pain level (P=0.0001). Lateral excursion movements significantly decreased (P=0.04). The occlusion remained stable in all cases. Complications occurred in six patients. Comparison analysis of the three groups demonstrated significantly better outcomes for patients with fewer previous TMJ surgeries and without exposure to Proplast-Teflon or Silastic TMJ implants. This study demonstrated that the TMJ Concepts/Techmedica total joint prosthesis is a viable technique for TMJ reconstruction as a primary procedure and for patients with previous multiple TMJ surgeries and mutilated anatomy of the TMJ. PMID- 12767874 TI - Evaluation of respiratory status and mandibular movement after total temporomandibular joint replacement in patients with rheumatoid arthritis. AB - We performed total TMJ replacement to improve respiratory status and correct occlusion in six patients with destruction of the temporomandibular joint (TMJ) caused by rheumatoid arthritis. Morphological changes were evaluated on lateral cephalograms before and after surgery. Respiratory function and mandibular movement were assessed with the use of an apnea-monitor and an LED mandibular tracking device, respectively. After surgery, symptoms such as snoring and daytime sleepiness improved, and solid food could be masticated. Postoperative cephalograms showed that both the posterior airway space and ramal height were significantly improved by surgery. Postoperative records of mandibular movement indicated stability of the occlusion and improvement of mandibular movement, as compared with the preoperative records. Mean oxygen saturation significantly improved 1 month after surgery, whereas apnea and apnea-hypopnea indices did not change significantly. PMID- 12767875 TI - Treatment of verrucous hyperplasia and verrucous carcinoma by shave excision and simple cryosurgery. AB - Wide excision and skin or mucosa grafting has been the primary form of therapy for verrucous hyperplasia and verrucous carcinoma. But those with wide involvement often make the procedures complicated. A simple management, shave excision followed by simple cryosurgery, is presented. The treatment modality described is simple, and less traumatic. It can be performed under local anaesthesia, and requires no hospitalization. It has been easily used to treat 26 lesions in 20 patients with satisfactory results. PMID- 12767876 TI - An analysis of cervical lymph nodes metastasis in oral squamous cell carcinoma. Relationship between grade of histopathological malignancy and lymph nodes metastasis. AB - Cervical lymph nodes metastasis is well known to be an indicator of poor prognosis in patients with oral cancer. This study was considered 38 patients of oral squamous cell carcinoma who were treated at our department from 1993 to 1997. Histological malignancy of initial biopsy in the pretreatment period was evaluated by Anneroth's classification. The relationship of degree of histological malignancy with cervical lymph nodes metastasis were analysed in this study. Metastasis in the cervical lymph nodes was histologically confirmed in 17 out of 38 cases (44.7%). The metastasis located predominantly in the submandibular nodes and superior internal jugular nodes. According to T classification, the rate of metastasis in the cervical lymph nodes in T1 and T2 cases was around 30%, while the rate in T3 and T4 cases tended to be as high as around 60%. However, there was no significant difference. A significant relation was evident between the degree of histological malignancy and metastasis in the cervical lymph nodes (P< 0.01), indicating that the histological malignancy could be served as a predictor for metastasis in the cervical lymph nodes. Among the six parameters, a significant difference was observed only in the degree of keratinization and the mode of invasion (P< 0.05). When the sum of the degree of histological malignancy exceeds 15, metastasis in the cervical lymph nodes should be considered. PMID- 12767877 TI - Osteoradionecrosis of the jaws as a side effect of radiotherapy of head and neck tumour patients--a report of a thirty year retrospective review. AB - This retrospective study aimed to demonstrate the incidence and the aetiological factors involved in osteoradionecrosis (ORN) in a group of 830 head and neck tumour patients who received radiotherapy between 1969 and 1999. The data showed an over all incidence of 8.2% and a 3-fold higher incidence for men than for women. Osteoradionecrosis was most commonly located in the body of the mandible. Concerning the risk factors, a negative influence was shown for advanced tumours, segmental resections of the mandible and pre-/post-radiation tooth extractions. Tooth extractions were found to be responsible for 50% of all cases. The osteoradionecroses were observed significantly earlier in patients who received pre-surgical radiotherapy than those who received post-surgical radiotherapy. Combined pre-surgical radio- and chemotherapy significantly hastened the appearance of osteoradionecrosis compared to pre-surgical radiotherapy alone. Only 40% of patients with osteoradionecrosis could be healed completely by means of surgery and antibiotic medication. Hyperbaric oxygenation (HBO) therapy was performed only in individual cases. The data suggest that osteoradionecrosis has a multifactorial aetiology. Therefore, a very close follow-up of tumour patients and a strict prophylactic management are required. PMID- 12767878 TI - Correlation of biomechanical stiffness with plain radiographic and ultrasound data in an experimental mandibular distraction wound. AB - The aim of this study was to determine the stiffness of a porcine mandibular distraction wound at the end of neutral fixation.Twenty-four Yucatan minipigs underwent unilateral mandibular distraction: zero-day latency, rates of 1, 2, and 4 mm per day, distraction gap of 12 mm. Radiographic and ultrasound bone fill scores at 0, 8, 16 and 24 days of neutral fixation were used to evaluate interval healing. At sacrifice, biomechanical stiffness was determined using an Instron machine to produce a downward force of 2 mm/min at the premolar occlusal surface. Force (kilo-Newton, kN) and displacement (mm) were recorded at a rate of 10,000 points/second. Stiffness across the distraction wound, plain radiographic and ultrasound bone fill scores all increased with duration of neutral fixation and were highest for the 1 mm per day group at all time points. At the end of fixation, even in the most clinically stable group, stiffness across the distraction wound was only 25.5% of control.Despite clinical stability and high bone fill scores by X-ray and ultrasound, the distraction wound did not achieve normal stiffness at the end of neutral fixation in this model. This may have implications for the management of patients who undergo large movements by distraction. PMID- 12767879 TI - Influence of donor age and culture conditions on tissue engineering of mucosa autografts. AB - In oral surgery the transplantation of tissue engineered mucosa is used more frequently. The conventional single cell suspension culture method (SCSM) involves murine feeder cells and foetal calf serum. The explant technique (ET) has been used as alternative culture procedure. Aim was to study the efficacy of the ET and the SCSM without feeder cells to grow primary cultures and to test the effects of donor age, of extracellular matrix proteins (ECMP), and of autogenous serum on cell growth in explant cultures. These factors were assessed in cultures of 58 patients overall. In 48 cultures of 12 patients primary cell growth was compared between the ET and the SCSM. Eighteen of 24 cultures were established with the ET whereas only 3 of 24 were established with the SCSM. To test the influence of donor age on cell multiplication, the proliferation rate (DNA synthesis measured by bromodeoxyuridine uptake) and the overall growth (DNA content) was determined in cultures of five young and five old donors. In cultures from old donors (mean age 56 years) proliferation was lower but more sustained relative to the cultures from the young donors (mean age 25 years). In old donors overall in vitro cell growth was only 2/3 of that in young donors. In cultures of 20 donors the influence on cell adhesion and growth of the ECMP fibronectin and laminin was assessed by planimetry. While ECMP augmented explant adhesion, these substances did not enhance keratinocyte growth significantly. Comparing the influence of autogenous and foetal calf serum on cell growth no differences were observed in all cultures of the six donors. In conclusion, the ET without additional ECMP coating and with autogenous instead of foetal calf serum are now used to culture gingival keratinocytes for tissue engineering mucosa grafts. Consequently xenogenous components are avoided, being a considerable advantage. PMID- 12767880 TI - Preclinical study of recombinant human bone morphogenetic protein-2: application of hyperbaric oxygenation during bone formation under unfavourable condition. AB - The objective of this study was to evaluate the effect of hyperbaric oxygenation on bone formation by recombinant human bone morphogenetic protein-2 (rhBMP-2) under unfavourable conditions. The calf muscles of 10 rats with low-blood supply were prepared by ligating and cutting the right femoral artery, and 10 micro of rhBMP-2 was implanted in the calf muscle. Five rats each were randomly assigned to the hyperbaric oxygenation group and the control group (untreated). The rats in the hyperbaric oxygenation group were treated with hyperbaric oxygenation at 2.0 atmospheres absolute for 3 weeks. In the histologic evaluation, the number of osteoblasts in the hyperbaric oxygenation group was greater than that in control group. The area of the trabecular bone induced in the hyperbaric oxygenation group was significantly larger than that in the control group. The values of alkaline phosphatase activity and calcium contents in the hyperbaric oxygenation group were significantly higher than those in the control group. The results of the present study suggest that hyperbaric oxygenation increases the partial oxygen pressure in low blood supply tissue and accelerates the activity and rate of osteoinduction by rhBMP-2. Hyperbaric oxygenation therapy may increase the clinical application of rhBMP-2 to unfavourable condition. PMID- 12767882 TI - An anatomical study of the cutaneous branches of the mental nerve. AB - Minor surgical procedures to the inner (mucosal) aspect of the lower lip may occasionally cause numbness of the overlying skin. This study was designed to find, by means of dissection and computerized three-dimensional reconstruction, why surgical interference with nerve fibres in the deep aspect of the lip can cause neurological deficit in the superficial layers. Thirteen cadaveric lips were examined by dissection under a surgical microscope (9 lips) or serial sectioning and computerized three-dimensional reconstruction (4 lips). Muscle mass, minor labial salivary glands and nerve fibres were identified and traced. Three patterns of mental nerve distribution were seen on dissection and two on computerized reconstruction; these latter corresponded to two of the patterns seen on dissection. Fibres passing close to the labial minor salivary gland mass were seen to travel towards the superficial aspect of the lip, terminating in the dermis. It is clear that there is no safe anatomical space for minor surgical procedures to the inner (mucosal) aspect of the lower lip if avoidance of cutaneous numbness is an important consideration. However, we describe a technique that may minimize the possibility of cutaneous numbness. PMID- 12767881 TI - Response of Schwann cells in the inferior alveolar nerve to distraction osteogenesis: an ultrastructural and immunohistochemical study. AB - The biological mechanisms of nerve adaptation to distraction osteogenesis have not yet been elucidated. This study observed response of Schwann cells in the inferior alveolar nerve (IAN) following mandibular lengthening by electron microscopy and immunohistochemistry of S-100 protein, a specific marker of Schwann cells. Unilateral mandibular distraction (10mm elongation) was performed in nine young adult goats. Three animals were sacrificed at 7, 14 and 28 days after completion of distraction, respectively. The distracted IAN specimens and control nerves (from the contralateral sides) were harvested and processed for histological, ultrastructural and immunohistochemical examinations. Wallerian degeneration was observed in the distracted IAN, and Signs of axonal regeneration, as well as many activated Schwann cells were seen in the lengthened nerves. The expression of S-100 protein increased significantly at early stage of distraction osteogenesis, but almost returned to the normal level at 28 days after distraction. This study suggests that Wallerian degeneration caused by mechanical stretching may stimulate Schwann cells to enter a proliferated and activated state. Schwann cells and S-100 protein appear to play crucial roles in axonal regeneration that contributes to nerve adaptation to gradual distraction. Therefore, the IAN injury caused by mandibular gradual distraction was not serious; it seems to recover totally through a complicated repair mechanism. PMID- 12767883 TI - Patient-centred outcome measures for oral mucosal disease are sensitive to treatment. AB - The aim of this study was to evaluate the sensitivity of two patient-centred outcome measures to the topical application of a corticosteroid (betamethasone) in the treatment of oral lichen planus (OLP). Forty-eight patients with clinical and histological features of OLP were recruited to take part in a 6-week study of the effectiveness of topical betamethasone for the treatment of symptomatic OLP. Participants completed a questionnaire incorporating the 16-item UK Oral Health Related Quality Of Life measure (OHQOL) and the 14-item Oral Health Impact Profile (OHIP-14), rated their pain on 'global' and visual analogue scales (VAS) and underwent an oral examination, at the start and end of the trial. Four (8?%) patients failed to complete the study. The clinical signs of OLP had improved for half (22) of the patients following treatment. Twenty-nine (66%) reported that their oral pain had reduced ('global' scale). More objectively, there were significant differences in VAS ratings of pain (P =0.005), OHIP-14 scores (P =0.036) and OHQOL scores (P =0.003) between the start and end of the trial. In conclusion, both OHQOL and OHIP-14, patient-centred outcome measures are sensitive to the clinical effects of topical betamethasone in the treatment of oral lichen planus. PMID- 12767884 TI - Use of a nasogastric catheter to prevent soft tissue entanglement of the externally ported distractor arm. AB - A simple method for the protection of the surrounding soft tissue during activation of a percutaneously ported distractor arm is presented. This reduces tissue damage, enhances patient compliance and could improve the overall success rate of the technique. PMID- 12767885 TI - Malignant myoepithelioma. Clinicopathological and immunohistochemical characteristics. AB - A malignant myoepithelioma arising in the submucosal accessory glands of the oral cavity of a 23-year-old woman is reported. The patient was very young compared to the cases in the literature, and the tumour had an unusual vestibular location. A false diagnosis of pleomorphic adenoma had been made by fine needle aspiration biopsy. The surgery comprised a wide tumour excision, but neck dissection was not indicated. The final histologic diagnosis was malignant myoepithelioma. Immunohistochemically putative myoepithelial markers were highly expressed. PMID- 12767886 TI - Extraskeletal myxoid chondrosarcoma of the infratemporal fossa: a case report. AB - Extraskeletal myxoid chondrosarcoma is a rare tumour affecting the head and neck. We present a new case located in the midfacial region. Clinical, pathological and therapeutical features are reviewed. PMID- 12767887 TI - Measurement of the dissociation constant of Fluo-3 for Ca2+ in isolated rabbit cardiomyocytes using Ca2+ wave characteristics. AB - The Ca(2+) dissociation constant (K(d)) of Fluo-3 was determined using confocal fluorescence microscopy in two different situations: (i) within the cytosol of a permeabilised cardiomyocyte; and (ii) in an intact cardiomyocyte after incubation with the acetoxymethyl ester form of Fluo-3 (AM). Measurements were made on isolated rabbit ventricular cardiomyocytes after permeabilisation by a brief treatment with beta-escin (0.1mg/ml) and equilibration with 10 microM Fluo-3. The K(d) of Fluo-3 within the cytosol was not significantly different from that in free solution (558 +/- 15 nM, n=6). Over a range of cytoplasmic [Ca(2+)], the minimum [Ca(2+)] values between Ca(2+) waves was relatively constant despite changes in wave frequency. After loading intact cardiomyocytes with Fluo-3 by incubation with the -AM, spontaneous Ca(2+) waves were produced by incubation with strophanthidin (10 microM). By assuming a common minimum [Ca(2+)] in permeabilised and intact cells, the intracellular K(d) of Fluo-3 in intact myocytes was estimated to be 898 +/-64 nM (n=6). Application of this K(d) to fluorescence records shows that Ca(2+) waves in intact cells have similar amplitudes to those in permeabilised cells. Stimulation of cardiac myocytes at 0.5 Hz in the absence of strophanthidin (room temperature) resulted in a Ca(2+) transient with a maximum and minimum [Ca(2+)] of 1190 +/- 200 and 158 +/- 30 nM (n=11), respectively. PMID- 12767888 TI - Ca2+ sparks induced by Na/Ca exchange. AB - Whether Ca(2+) influx on the Na/Ca exchanger (NCX) can trigger elementary sarcoplasmic reticulum (SR) Ca(2+) release events (Ca(2+) sparks) is controversial. We imaged [Ca(2+)](i) (Nipkow confocal microscope and fluo-3) in left ventricular myocytes isolated from wild type (WT) and transgenic (TG) mice overexpressing NCX 2.5-fold. Sudden activation of Ca(2+) influx via NCX induced by abrupt exposure to "0" [Na(+)](o)/normal [Ca(2+)](o) solution by means of a rapid solution switcher-induced Ca(2+) sparks in NCX TG myocytes in 425+/-17 ms, n=21. The diameter and amplitude (F/F(0)) of these sparks (2.74+/-0.14 microm, F/F(0)=2.16+/-0.06, n=18) were similar to those induced by field stimulation of myocytes in the presence of 20 microM nifedipine (2.70+/-0.10 microm, F/F(0)=1.98+/-0.08, n=17). In WT myocytes no Ca(2+) sparks were observed within the first 600 ms after abrupt removal of extracellular Na. In parallel experiments, voltage clamp current measurements (-80 mV) showed that the Na/Ca exchange current (I(NCX)) began within 60 ms of activation of the switcher, and peaked at 312+/-57 pA in TG myocytes within 300-500 ms. I(Ca,L) in 20 microM nifedipine was 10.3+/-4.3 pA, n=7. These results indicate that Ca(2+) entering the myocyte via NCX can cause Ca(2+) sparks which are similar to those elicited by electrical stimulation. However, Ca(2+) influx on NCX is much less efficient in inducing Ca(2+) sparks than Ca(2+) influx via I(Ca,L). PMID- 12767889 TI - Cardiac excitation-contraction coupling in the absence of Na(+) - Ca2+ exchange. AB - We investigate cardiac excitation-contraction coupling in the absence of sarcolemmal Na(+) - Ca(2+) exchange using NCX1 knock out mice. Knock out of NCX1 is embryonic lethal, and we measure Ca(2+) transients and contractions in heart tubes from embryos at day 9.5 post coitum. Immunoblot and electron microscopy both indicate that sarcoplasmic reticular membranes are diminished in the knock out (NCX(-/-)) heart tubes. Both Ni(2+) and nifedipine block excitation contraction coupling in NCX-containing (NCX+) and NCX(-/-) heart tubes indicating an essential role for the L-type Ca(2+) current. Under basal conditions (1Hz stimulation), the NCX(-/-) heart tubes have normal Ca(2+) transients but are unable to maintain homeostasis when Ca(2+) fluxes are increased by various interventions (increased stimulation frequency, caffeine, isoproterenol). In each case, the NCX(-/-) heart tubes respond to the intervention in a more deleterious manner (increased diastolic Ca(2+), decreased Ca(2+) transient) than the NCX+ heart tubes. Expression of the sarcolemmal Ca(2+) pump was not upregulated. The sarcolemmal Ca(2+) pump, however, was able to compensate surprisingly well for the absence of Na(+) - Ca(2+) exchange under basal conditions. PMID- 12767890 TI - A new technique for simultaneous and in situ measurements of Ca2+ signals in arteriolar smooth muscle and endothelial cells. AB - We report here the first local and global Ca(2+) measurements made from in situ terminal arterioles. The advantages of the method are that there is minimal disturbance to the vessels, which retain their relationship to the tissue they are supplying (rat ureter) and the small size of vessel that can be studied. Good loading with the Ca(2+) indicator, Fluo-4 was obtained, and confocal sectioning through the tissue enabled vascular smooth muscle and endothelial cells to be clearly seen, along with red blood cells, nerve endings and the ureteric smooth muscle cells. We find the terminal arterioles to be extremely active, both spontaneously and in response to nor-adrenaline stimulation, with Ca(2+) sparks occurring in the vascular myocytes and Ca(2+) puffs in the endothelial cells. Even under resting conditions, endothelial cells produced oscillations and waves, which could pass from cell to cell, whereas the vascular myocytes only produced waves in response to agonist stimulation, and with no increase in the frequency of Ca(2+) sparks, and no spread from cell to cell. We compare our data to those obtained in dissected intact vessels and single cells. We conclude that this approach is a convenient and useful method for studying inter- and intracellular Ca(2+) signalling events and communication between cell types, particularly in very small vessels. PMID- 12767891 TI - Ca(2+)-dependent and Ca2+ release-dependent excitation in leech photoreceptors: evidence from a novel "inside-out" cell model. AB - We have developed a novel, electrophysiologically intact and light-sensitive "inside-out" cell model (IOCM) of microvillar photoreceptors of the leech Hirudo medicinalis. Light responses recorded from the IOCM with sharp microelectrodes are depolarizations with amplitudes of up to 50-60 mV. In darkness, graded elevations of the free Ca(2+) concentration in the "intracellular medium" (ICM) reversibly increase the conductance of the microvillar membrane leading to Ca(2+) induced graded voltage changes up to approximately 50 mV. The threshold for Ca(2+)-induced voltage changes is approximately 0.06 microM, EC(50) is approximately 1.2 microM, and saturation occurs at approximately 20 microM free Ca(2+). Small Ca(2+) elevations (<0.6 microM) produce discrete waves of depolarization resembling quantum bumps. Stimulating IOCMs with short (20-ms) and long (5-s) light stimuli produces transient light responses (repolarization within ca. 200 ms) in an ICM containing only 10nM free Ca(2+). At 0.44 microM free Ca(2+) in the ICM, the microvillar membrane depolarizes by 10-20 mV and responses to 5-s light steps have an initial transient component and a plateau component, similar to responses in intact cells. Generation of the plateau component in IOCMs is suppressed by heparin and cyclopiazonic acid (CPA), agents that block inositol 1,4,5-trisphosphate (Ins(1,4,5)P(3))-induced Ca(2+) release from and Ca(2+) uptake into the endoplasmic reticulum (ER). These results indicate that there is a Ca(2+)-dependent conductance in the microvillar membrane and that the light-induced Ins(1,4,5)P(3)- and Ca(2+) release-mediated intracellular Ca(2+) elevation in leech photoreceptors contributes to the generation of the receptor potential, particularly the plateau component of responses to long steps of light. PMID- 12767892 TI - Synchronization of pancreatic beta-cell rhythmicity after glucagon induction of Ca2+ transients. AB - Pancreatic beta-cells are biological oscillators requiring a coupling force for the synchronization of the cytoplasmic Ca(2+) oscillations responsible for pulsatile insulin release. Testing the idea that transients, superimposed on the oscillations, are important for this synchronization, the concentration of cytoplasmic Ca(2+) ([Ca(2+)](i)) was measured with ratiometric fura-2 technique in single beta-cells and small aggregates prepared from islets isolated from ob/ob-mice. Image analyses revealed asynchronous [Ca(2+)](i) oscillations in adjacent beta-cells lacking physical contact. The addition of glucagon stimulated the firing of [Ca(2+)](i) transients, which appeared in synchrony in adjacent beta-cells. Moreover, the presence of glucagon promoted synchronization of the [Ca(2+)](i) oscillations in beta-cells separated by a distance <100 microm but not in those >200 microm apart. The results support the proposal that the repolarizing effect of [Ca(2+)](i) transients provides a coupling force for co ordinating the pulses of insulin release generated by pancreatic beta-cells. PMID- 12767893 TI - Cytoplasmic calcium measurement in rotavirus enterotoxin-enhanced green fluorescent protein (NSP4-EGFP) expressing cells loaded with Fura-2. AB - The green fluorescent protein (GFP) and its analogs are standard markers of protein expression and intracellular localization of proteins. The fluorescent properties of GFP complicate accurate measurement of intracellular calcium using calcium sensitive fluorophores, which show a great degree of spectral overlap with GFP, or their K(d) values are too high for accurate measurement of subtle changes in cytoplasmic calcium concentrations. Here we describe a simple modification of the standard microscope-based Fura-2 calcium-imaging technique which permits the quantitative measurement of intracellular calcium levels in cells expressing enhanced green fluorescent protein (EGFP) fusion proteins. Longpass emission filtering of the Fura-2 signal in cells expressing an EGFP fusion protein is sufficient to eliminate the EGFP-Fura-2 emission spectra overlap and allows quantitative calibration of intracellular calcium. To validate this technique, we investigated the ability of rotavirus enterotoxin NSP4-EGFP to elevate intracellular calcium levels in mammalian HEK 293 cells. We show here that inducible intracellular expression of NSP4-EGFP fusion protein elevates basal intracellular calcium more than two-fold by a phospholipase C (PLC) independent mechanism. PMID- 12767894 TI - Effects of peroxynitrite on pig coronary artery smooth muscle. AB - We examined the effects of peroxynitrite pretreatment of pig coronary arteries on their sarcoplasmic reticulum (SR) Ca(2+) pump function. Pretreating rings from de endothelialized arteries with peroxynitrite, followed by a wash to remove this agent, led to a decrease in the force of contraction produced in response to the SR Ca(2+) pump inhibitor cyclopiazonic acid (CPA, IC(50) = 87 +/- 6 microM). Inclusion of catalase and superoxide dismutase with the peroxynitrite did not alter its effect indicating that the inhibition was produced by peroxynitrite. Contractions produced by 30 mM KCl were not affected by up to 250 microM peroxynitrite. Smooth muscle cells cultured from this artery gave a transient increase in cytosolic Ca(2+) in response to CPA. Treating the cells with peroxynitrite inhibited this increase. Treating the SR-enriched isolated subcellular membrane fraction with peroxynitrite produced an inhibition of the ATP-dependent azide-insensitive oxalate-stimulated Ca(2+) uptake. Thus, peroxynitrite damages the SR Ca(2+)pump in the coronary artery, and this inhibition appears to lead to an inability of the arteries to respond to CPA. Thus, peroxynitrite produced from superoxide and NO in the arteries may compromise regulation of coronary tone which requires mobilization of Ca(2+) from the SR. PMID- 12767895 TI - Mechanisms of ATP-induced calcium signaling and growth arrest in human prostate cancer cells. AB - This study investigates the calcium mechanisms involved in growth arrest induced by extracellular ATP in DU-145 androgen-independent human prostate cancer cells. Exposure of DU-145 cells to 100 microM ATP produced an increase in cytoplasmic calcium concentration ([Ca(2+)](i)), due to a mobilization of calcium from the endoplasmic reticulum stores and to subsequent capacitative calcium entry (CCE). We have shown that this [Ca(2+)](i) increase occurs after stimulation by ATP of the phospholipase C (PLC) pathway. For the first time, we have identified the inositol 1,4,5-trisphosphate receptor (IP(3)R) isoforms expressed in this cell line and have demonstrated a participation of protein kinase C in CCE. Using fluorescence imaging, we have shown that a long-term treatment with ATP leads to a decrease in the intraluminal endoplasmic reticulum calcium concentration as well as in the amount of releasable Ca(2+). Modulating extracellular free calcium concentrations indicated that variations in [Ca(2+)](i) did not affect the ATP induced growth arrest of DU-145 cells. However, treating cells with 1 nM thapsigargin (TG) to deplete intracellular calcium pools prevented the growth arrest induced by ATP. Altogether, these results indicate that growth arrest induced in DU-145 cells by extracellular ATP is not correlated with an increase in [Ca(2+)](i) but rather with a decrease in intracellular calcium pool content. PMID- 12767896 TI - Refilling of cortical calcium stores in Paramecium cells: in situ analysis in correlation with store-operated calcium influx. AB - This is the first thorough study of refilling of a cortical calcium store in a secretory cell after stimulation in which we combined widely different methodologies. Stimulation of dense-core vesicle ("trichocysts") exocytosis in Paramecium involves a Ca(2+) -influx" superimposed to Ca(2+) -release from cortical stores ("alveolar sacs" (ASs)). In quenched-flow experiments, membrane fusion frequency rose with increasing [Ca(2+)](o) in the medium, from approximately 20-25% at [Ca(2+)](o) < or = 0.25 microM to 100% at [Ca(2+)](o) between 2 and 10 microM, i.e. close to the range of estimated local intracellular [Ca(2+)] during membrane fusion. Next, we analyzed Ca(2+)-specific fluorochrome signals during stimulation under different conditions. Treatment with actin reactive drugs had no effect on Ca(2+) -signaling. In double trigger experiments, with BAPTA in the second secretagogue application (BAPTA only for stimulation and analysis), the cortical Ca(2+) -signal (due solely to Ca(2+) released from cortical stores) recovered with t(1/2) approximately 65 min. When ASs were analyzed in situ by X-ray microanalysis after different trigger times (+Ca(2+)(o)), t(1/2) for store refilling was similar, approximately 60 min. These values are similar to previously measured 45Ca(2+) -uptake by isolated ASs. In sum we find, (i) exogenous Ca(2+) increases exocytosis/membrane fusion performance with EC(50)=0.7 microM, (ii) Ca(2+) -signaling in this system is not sensitive to actin-reactive drugs, and (iii) refilling of these cortical calcium stores goes on over hours and thus is much slower than expected. PMID- 12767897 TI - 2-Aminoethoxydiphenyl borate (2-APB) antagonises inositol 1,4,5-trisphosphate induced calcium release, inhibits calcium pumps and has a use-dependent and slowly reversible action on store-operated calcium entry channels. AB - The action of 2-aminoethoxydiphenyl borate (2-APB) on Ca(2+) signalling in HeLa cells and cardiac myocytes was investigated. Consistent with other studies, we found that superfusion of cells with 2-APB rapidly inhibited inositol 1,4,5 trisphosphate (InsP(3))-mediated Ca(2+) release and store-operated Ca(2+) entry (SOC). In addition to abrogating hormone-evoked Ca(2+) responses, 2-APB could antagonise Ca(2+) signals evoked by a membrane permeant InsP(3) ester. 2-APB also slowed the recovery of intracellular Ca(2+) signals consistent with an effect on Ca(2+) ATPases. The inhibitory action of 2-APB on InsP(3) receptors (InsP(3)Rs), SOC channels and Ca(2+) pumps persisted for several minutes after washout of the compound. Application of 2-APB to unstimulated cells had no effect on subsequent Ca(2+) responses suggesting that it has a use-dependent action. Mitochondria in cells treated with 2-APB showed a rapid and slowly reversible swelling. 2-APB did not cause the mitochondria to depolarise, but it reduced the extent of mitochondrial calcium uptake. Although 2-APB has been demonstrated not to affect voltage-operated Ca(2+) channels or ryanodine receptors, we found that it gave a concentration-dependent long-lasting inhibition of Ca(2+) signalling in electrically-stimulated cardiac myocytes, where InsP(3)Rs and SOC channels do not play a significant role. Our data suggest that 2-APB has multiple cellular targets, a use-dependent action, is difficult to reverse and may affect Ca(2+) signalling in cell types where InsP(3) and SOC are not active. PMID- 12767898 TI - Hidden intermediates and levinthal paradox in the folding of small proteins. AB - It has long been suggested that existence of partially folded intermediates may be essential for proteins to fold in a biologically meaningful time scale. Although partially folded intermediates have been commonly observed in larger proteins, they are generally not detectable in the kinetic folding of smaller proteins (approximately 100 amino acids or less). Recent native-state hydrogen exchange studies suggest that partially folded intermediates may exist behind the rate-limiting transition state in small proteins and evade detection by conventional kinetic methods. PMID- 12767899 TI - TNF-related weak inducer of apoptosis receptor, a TNF receptor superfamily member, activates NF-kappa B through TNF receptor-associated factors. AB - TNF-related weak inducer of apoptosis (TWEAK) is a member of the TNF ligand family that induces angiogenesis in vivo. The TWEAK receptor (TweakR) is a recently identified member of the TNF receptor (TNFR) superfamily and is expressed on smooth muscle cells (SMCs) and endothelial cells (ECs). In this report we identify the TNF receptor-associated factor (TRAF) family of signal transducers as important components of TweakR-mediated NF-kappa B activation. Coimmunoprecipitation experiments suggested potential interactions between the cytoplasmic tail of TweakR with TRAFs 1, 2, 3, and 5. Dominant negative forms of TRAF2 and TRAF5 substantially inhibited TweakR-mediated NF-kappa B activation, suggesting a role of TRAFs in regulating smooth muscle and endothelial cell function. Using alanine-scanning analysis, we defined a TRAF-binding motif, PIEET, in TweakR that mediates TRAF binding and NF-kappa B activation. Furthermore, TweakR mutations within the TRAF-binding motif abolished TweakR stimulated SMC migration, revealing a role for TRAFs in TweakR-induced activation events. PMID- 12767901 TI - Cdc42 plays a critical role in assembly of sarcomere units in series of cardiac myocytes. AB - Cardiomyocyte hypertrophy is observed in various cardiovascular diseases and causes heart failure. We here examined the role of small GTP-binding proteins of Rho family in phenylephrine (PE)-or leukocyte inhibitory factor (LIF)-induced hypertrophic morphogenesis of cultured neonatal rat cardiomyocytes. Both LIF and PE increased cell size of cardiomyocytes. LIF induced an increase in the length/width ratio of cardiomyocytes, while PE did not change the ratio. Adenoviral gene transfer of constitutively active mutants of Cdc42 increased the length/width ratio of cardiomyocytes and dominant negative mutants of Cdc42 conversely inhibited LIF-induced cell-elongation, while mutants of RhoA and Rac1 did not affect the length/width ratio of cardiomyocytes. These results suggest that Cdc42, but not RhoA and Rac1, is involved in LIF-induced sarcomere assembly in series in cardiomyocytes. PMID- 12767900 TI - Inhibition of interferon-gamma-activated nuclear factor-kappa B by cyclosporin A: A possible mechanism for synergistic induction of apoptosis by interferon-gamma and cyclosporin A in gastric carcinoma cells. AB - We previously reported synergistic induction of apoptosis by IFN-gamma plus either cyclosporin A (CsA) or tacrolimus (FK506) in gastric carcinoma cells. In this study, we aimed to elucidate the mechanism for this synergistic induction of apoptosis. IFN-gamma plus CsA synergistically induced caspase-3 mediated apoptosis in gastric carcinoma cells. Although IFN-gamma induced activation of signal transducer and activator of transcription1 (STAT1) and expression of interferon regulatory factor-1 (IRF-1) mRNA, IFN-gamma alone was not able to induce caspase-3 activation and apoptosis. When gastric carcinoma cells were treated with cyclohexamide, a protein synthesis inhibitor, following IFN-gamma pretreatment, caspase-3 was activated, and apoptosis was markedly induced. These findings suggest the existence of IFN-gamma-induced anti-apoptotic pathway and we evaluated the effect of IFN-gamma and CsA on calcium-sensitive nuclear factor kappa B (NF-kappa B) activation. IFN-gamma increased intracellular calcium ion concentration ([Ca(2+)](i)) consisting of a spike and a sustained phase, and the latter was completely abrogated by CsA. Activation of NF-kappa B occurred in response to IFN-gamma, and which was markedly inhibited by either CsA or FK506. NF-kappa B decoy also enhanced the cytotoxic effect of IFN-gamma. These results suggest that IFN-gamma may simultaneously induce the STAT1-mediated apoptotic pathway and the anti-apoptotic pathway through calcium-activated NF-kappa B and that inhibition of the latter by CsA may result in dominance of the apoptosis inducing pathway. PMID- 12767902 TI - C-chain-bound glycogenin is released from proteoglycogen by isoamylase and is able to autoglucosylate. AB - Proteoglycogen glycogenin is linked to the glucose residue of the C-chain reducing end of glycogen. We describe for the first time the release by isoamylase and isolation of C-chain-bound glycogenin (C-glycogenin) from proteoglycogen. The treatment of proteoglycogen with alpha-amylase releases monoglucosylated and diglucosylated glycogenin (a-glycogenin) which is able to autoglucosylate. It had been described that isoamylase splits the glucose glycogenin linkage of fully autoglucosylated glycogenin previously digested with trypsin, releasing the maltosaccharide moiety. It was also described that carbohydrate-free apo-glycogenin shows higher mobility in SDS-PAGE and twice the autoglucosylation capacity of partly glucosylated glycogenin. On the contrary, we found that the C-glycogenin released from proteoglycogen by isoamylolysis shows lower mobility in SDS-PAGE and about half the autoglucosylation acceptor capacity of the partly glucosylated a-glycogenin. This behavior is consistent with the release of maltosaccharide-bound glycogenin instead of apo-glycogenin. No label was split from auto-[14C]glucosylated C-glycogenin or fully auto [14C]glucosylated a-glycogenin subjected to isoamylolysis without previous trypsinolysis, thus proving no hydrolysis of the maltosaccharide-tyrosine linkage. The ability of C-glycogenin for autoglucosylation would indicate that the size of the C-chain is lower than the average length of the other glycogen chains. PMID- 12767903 TI - Blocking of alpha 5 integrin stimulates production of TGF-beta and PAI-1 by human mesangial cells. AB - Expression of integrin, which mediates cell-matrix interaction, is affected by several cytokines, in particular by transforming growth factor-beta (TGF-beta). However, it is unknown whether, in an opposite way, a specific integrin is involved in cytokine synthesis. We tested this hypothesis. Function-blocking anti alpha 5 integrin (fibronectin receptor) antibody increased TGF-beta secretion in growth-arrested human mesangial cells (2.3-fold) compared with control IgG or anti-alpha v beta 3 integrin (receptor for several matrix proteins) antibody. It also increased the secretion of plasminogen activator inhibitor-1 (PAI-1), a protein associated with matrix increase, by 3.2-fold. The increase in PAI-1 secretion induced by anti-alpha 5 integrin antibody was not abrogated by anti-TGF beta neutralizing antibody. These results indicate that function-blocking of anti alpha 5 integrin stimulates TGF-beta as well as PAI-1 production, suggesting that alpha 5 integrin is involved in fibrotic process. Function-modulation of a specific integrin thus appears to play a role in glomerular remodeling. PMID- 12767904 TI - Synthesis and secretion of transferrin by isolated ciliary epithelium of rabbit. AB - It has been shown that the vitreous contains several intrinsic glycoproteins whose origin remains to be clarified. Isolated ciliary epithelium (CE) was assayed to verify its role in the synthesis and secretion of transferrin for the vitreous body. It was cultured in the presence of [35S]methionine and the incubation medium was processed for immunoprecipitation. Total RNA from CE was processed for RT-PCR and the amplification products were sequenced. Also, whole preparations of isolated CE were processed for immunolocalization of transferrin. From the incubation assays, a labeled peptide of about 80 kDa was immunopurified that is the expected size of transferrin. The RT-PCR and sequencing experiments detected the presence of transferrin mRNA. Both layers of the CE exhibited transferrin reactivity, following immunohistochemical processing. Taken altogether, these results indicate the CE as one of the possible sources of vitreous intrinsic transferrin. PMID- 12767905 TI - Overexpression of glycogen synthase in mouse muscle results in less branched glycogen. AB - Glycogen, a branched polymer of glucose, serves as an energy reserve in many organisms. The degree of branching likely reflects the balance between the activities of glycogen synthase and branching enzyme. Mice overexpressing constitutively active glycogen synthase in skeletal muscle (GSL30) have elevated muscle glycogen. To test whether excess glycogen synthase activity affected glycogen branching, we examined the glycogen from skeletal muscle of GSL30 mice. The absorption spectrum of muscle glycogen determined in the presence of iodine was shifted to higher wavelengths in the GSL30 animals, consistent with a decrease in the degree of branching. As judged by Western blotting, the levels of glycogenin and the branching enzyme were also elevated. Branching enzyme activity also increased approximately threefold. However, this compared with an increase in glycogen synthase of some 50-fold, so that the increase in branching enzyme in response to overexpression of glycogen synthase was insufficient to synthesize normally branched glycogen. PMID- 12767906 TI - Insulin-like growth factor-1 downregulates nuclear factor kappa B activation and upregulates interleukin-8 gene expression induced by tumor necrosis factor alpha. AB - Pretreatment of HT29-D4 epithelial adenocarcinoma colic cells with des-IGF-1 upregulated TNF alpha-mediated activation of IL-8 expression at different levels (protein, mRNA, and hnRNA). RNA transcription but not RNA stabilization was found to be involved. In this cell line, cooperation of NF-kappa B with other factors appeared essential for IL-8 expression. Indeed, TNF alpha-induced NF-kappa B translocation was not sufficient to support enhancement of the transcription and des-IGF-1 did not promote but partly inhibited both the TNF alpha-induced NF kappa B activation and I kappa B alpha degradation through a PI-3K-dependent pathway. A CCAAT/enhancer binding protein (C/EBP) site located on the IL-8 gene enhancer cooperated with a NF-kappa B binding site and led to the upregulation of IL-8 expression. Binding of C/EBP alpha to this sequence disappeared in IGF-1 treated cells. This event may be important for the cross-talk between IGF-1- and TNF alpha-mediated pathways leading to the control of inflammatory processes and the decision concerning apoptosis or cell survival. PMID- 12767907 TI - House fly cytochrome b5 exhibits kinetically trapped hemin and selectivity in hemin binding. AB - We report that cytochrome b(5) (cyt b(5)) from Musca domestica (house fly) is more thermally stable than all other microsomal (Mc) cytochromes b(5) that have been examined to date. It also exhibits a much higher barrier to equilibration of the two isomeric forms of the protein, which differ by a 180 degrees rotation about the alpha-gamma-meso axis of hemin (ferric heme). In fact, hemin is kinetically trapped in a nearly statistical 1.2:1 ratio of rotational forms in freshly expressed protein. The equilibrium ratio (5.5:1) is established only upon incubation at temperatures above 37 degrees C. House fly Mc cyt b(5) is only the second b-hemoprotein that has been shown to exhibit kinetically trapped hemin at room temperature or above, the first being cyt b(5) from the outer membrane of rat liver mitochondria (rat OM cyt b(5)). Finally, we show that the small excess of one orientational isomer over the other in freshly expressed protein results from selective binding of hemin by the apoprotein, a phenomenon that has not heretofore been established for any apocyt b(5). PMID- 12767908 TI - Posttranscriptional regulation of TSC-22 (TGF-beta-stimulated clone-22) gene by TGF-beta 1. AB - TSC-22 gene was composed of three exons and its length was approximately 5.5 kb including 2.9 kb promoter region. The transcription starting site was located at 7 and 29 bp downstream from TATA box. Promoter analysis revealed that 2146 bp of TSC-22 promoter was activated by several differentiation inducing drugs. Although originally TSC-22 was isolated as a TGF-beta-inducible gene, TSC-22 promoter was not activated by the enhanced TGF-beta signaling. We found 3 copies of the Shaw Kamens sequence (AUUUA) in the human TSC-22 mRNA 3'-UTR and identified three proteins (40, 20, and 15 kDa) which bound to this. Only the 40 kDa protein-RNA complex was decreased by treatment with TGF-beta 1. Moreover, the TSC-22 mRNA 3' UTR destabilized the heterologous luciferase mRNA, but the destabilization was recovered with TGF-beta 1. These observations suggest that up-regulation of TSC 22 mRNA by TGF-beta 1 is achieved by mRNA stabilization, but not by transcriptional activation. PMID- 12767909 TI - Characterization of a cAMP-dependent protein kinase catalytic subunit from rainbow trout spermatozoa. AB - The cyclic AMP-dependent phosphorylation of proteins is essential for the initiation of sperm motility in salmonid fishes. This study isolated cDNA for the catalytic subunit of a cAMP-dependent protein kinase (PKA-C) from rainbow trout testis. The deduced amino acid sequence shows 75-80% identity to sequences previously reported in other organisms. However, the N-terminal regions of PKA-C from the testis as well as ovary in the trout appear slightly shorter than those from other tissues, suggesting that small PKA-C might be specific to germ cells. An immunofluorescence study using polyclonal antibody against trout testis PKA-C shows that it localizes along sperm flagellum. Furthermore, immunoelectron microscopy revealed that PKA-C is anchored to the outer arm dynein of flagellar axonemes. These results suggest that PKA-C is involved in regulating the flagellar motility of sperm via phosphorylation of a subunit of the outer arm dynein. PMID- 12767910 TI - Oxygen-evolving enhancer protein 2 is phosphorylated by glycine-rich protein 3/wall-associated kinase 1 in Arabidopsis. AB - The Arabidopsis wall-associated receptor kinase, WAK1, is a member of WAK family that links the plasma membrane to the extracellular matrix. A glycine-rich secreted protein, AtGRP-3, was previously shown to regulate WAK1 functions through binding to the extracellular domain of WAK1. In this study, we sought to determine the downstream molecules of the AtGRP-3/WAK1 signaling pathway, by using two-dimensional gel electrophoresis combined with Edman sequencing and matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS). We report here that a chloroplast protein, oxygen-evolving enhancer protein 2 (OEE2), specifically interacts with the cytoplasmic kinase domain of WAK1 and becomes phosphorylated in an AtGRP-3-dependent manner. The phosphorylation of OEE2 is also induced in Arabidopsis by treatment with avirulent Pseudomonas syringae. Taken together, these results suggest that OEE2 activity is regulated by AtGRP-3/WAK1. PMID- 12767911 TI - Cloning of a salivary gland metalloprotease and characterization of gelatinase and fibrin(ogen)lytic activities in the saliva of the Lyme disease tick vector Ixodes scapularis. AB - The full-length sequence of tick salivary gland cDNA coding for a protein similar to metalloproteases (MP) of the reprolysin family is reported. The Ixodes scapularis MP is a 488 amino acid (aa) protein containing pre- and pro-enzyme domains, the zinc-binding motif HExxHxxGxxH common to metalloproteases, and a cysteine-rich region. In addition, the predicted amino-terminal sequences of I. scapularis MPs were found by Edman degradation of PVDF-transferred SDS/PAGE separated tick saliva proteins, indicating that these putative enzymes are secreted. Furthermore, saliva has a metal-dependent proteolytic activity towards gelatin, fibrin(ogen), and fibronectin, but not collagen or laminin. Accordingly, I. scapularis saliva has a rather specific metalloprotease similar to the hemorrhagic proteases of snake venoms. This is the first description of such activity in tick saliva and its role in tick feeding and Borrelia transmission is discussed. PMID- 12767912 TI - Heme oxygenase-1 fused to a TAT peptide transduces and protects pancreatic beta cells. AB - Transplantation of islets is becoming an established method for treating type 1 diabetes. However, viability of islets is greatly affected by necrosis/apoptosis induced by oxidative stress and other insults during isolation and subsequent in vitro culture. Expression of cytoprotective proteins, such as heme oxygenase-1 (HO-1), reduces the deleterious effects of oxidative stress in transplantable islets. We have generated a fusion protein composed of HO-1 and TAT protein transduction domain (TAT/PTD), an 11-aa cell penetrating peptide from the human immunodeficiency virus TAT protein. Transduction of TAT/PTD-HO-1 to insulin producing cells protects against TNF-alpha-mediated cytotoxicity. TAT/PTD-HO-1 transduction to islets does not impair islet physiology, as assessed by reversion of chemically induced diabetes in immunodeficient mice. Finally, we report that transduction of HO-1 fusion protein into islets improves islet viability in culture. This approach might have a positive impact on the availability of islets for transplantation. PMID- 12767913 TI - In vitro bone formation using muscle-derived cells: a new paradigm for bone tissue engineering using polymer-bone morphogenetic protein matrices. AB - Over 800,000 bone grafting procedures are performed in the United States annually, creating a demand for viable alternatives to autogenous bone, the grafting standard in osseous repair. The objective of this study was to examine the efficacy of a BMP-polymer matrix in inducing the expression of the osteoblastic phenotype and in vitro bone formation by muscle-derived cells. Specifically, we evaluated the ability of bone morphogenetic protein-7 (BMP-7), delivered from a poly(lactide-co-glycolide) (PLAGA) matrix, to induce the differentiation of cells derived from rabbit skeletal muscle into osteoblast-like cells and subsequently form mineralized tissue. Results confirmed that muscle derived cells attached and proliferated on the PLAGA substrates. BMP-7 released from PLAGA induced the muscle-derived cells to increase bone marker expression and form mineralized cultures. These results demonstrate the efficacy of a BMP polymer matrix in inducing the expression of the osteoblastic phenotype by muscle derived cells and present a new paradigm for bone tissue engineering. PMID- 12767914 TI - Ceramide inhibits the potassium channel Kv1.3 by the formation of membrane platforms. AB - Previous studies suggested a central role of sphingomyelin- and cholesterol enriched membrane rafts in the initiation of signaling via many receptors. Here, we investigated the role of membrane rafts for the function of the voltage-gated potassium channel Kv1.3. We demonstrate that Kv1.3 localizes in the cell membrane to pre-existing small, sphingolipid- and cholesterol-enriched membrane rafts. Transformation of these small rafts to large ceramide-enriched membrane platforms was achieved by stimulation of the endogenous acid sphingomyelinase, addition of exogenous sphingomyelinase or treatment of the cells with C(16)-ceramide and resulted in clustering of Kv1.3 within ceramide-enriched membrane platforms and inhibition of the channel's activity. Likewise, disruption of pre-existing small rafts inhibited Kv1.3 activity. This indicates that intact small membrane rafts are required for Kv1.3 activity and an alteration of the lipid environment of rafts inhibits Kv1.3. These data, thus, may suggest a novel concept for the regulation of ion channels by the cell membrane composition. PMID- 12767915 TI - Effects of endothelin-1 on mitochondrial function during the protection against myocardial cell apoptosis. AB - Endothelin-1 is a potent survival factor against myocardial cell apoptosis. While apoptotic stimuli often perturb mitochondrial function by decreasing the membrane potential as well as oxygen consumption, it is unknown whether ET-1 can rescue such perturbation by apoptotic stimuli. Administration of endothelin-1 inhibited the H(2)O(2)-induced release of cytochrome c from mitochondria to the cytosol in cardiac myocytes, indicating the involvement of the mitochondria-dependent pathway in the anti-apoptotic effect of endothelin-1. We showed here by cytofluorimetric analysis that endothelin-1 prevented the H(2)O(2)-induced decrease of membrane potential. However, endothelin-1 was unable to reverse the H(2)O(2)-mediated decrease in oxygen consumption and electron transport in the mitochondria of cardiac myocytes. Endothelin-1 was unable to rescue cardiac myocytes from apoptosis when administered after the decrease in mitochondrial membrane potential. These data suggest that endothelin-1 does not target the mitochondrial respiratory chain, but rather stabilizes the mitochondrial membrane during the protection against apoptosis. PMID- 12767916 TI - Establishment of lacZ-transgenic rats: a tool for regenerative research in myocardium. AB - Animals transgenic (Tg) for reporter genes would be useful to following a given cell lineage during differentiation and regeneration processes. Here, we established a beta-galactosidase (lacZ) Tg rat to use as a tool for regenerative research. Strong lacZ expression was observed in the skeletal muscles, myocardium, pancreas, and skin obtained from these lacZ-Tg rats, and moderate lacZ expression was observed in the liver, spleen, kidney, and cartilage. In contrast, brain, vessels, lung, adrenal gland, small intestine, blood leukocytes, bone marrow (BM) cells, and peripheral blood cells showed no lacZ expression. To test whether this lacZ-Tg rat could be used for regenerative research in myocardium, we induced myocardial injury after a lacZ-Tg BM transplant (BMT) into wild-type rats. The results show that lacZ-positive cardiomyocytes were found in the peri-infarct and uninjured myocardium in the BMT recipient rats. These findings suggest that lacZ-Tg rats are useful tool for regenerative research in the myocardium. PMID- 12767917 TI - Inhibition of carbonic anhydrase II by steroidal and non-steroidal sulphamates. AB - Carbonic anhydrases (CAs) are expressed by many solid tumours where they may act to confer a growth advantage on malignant tissues. In this study we have examined the ability of a series of steroidal and non-steroidal sulphamates (originally developed as steroid sulphatase inhibitors) and related compounds to inhibit human CAII (hCAII) activity in vitro. Using a 96-well plate assay, oestrone-3-O sulphamate (EMATE) and two coumarin-based sulphamate drugs (667 COUMATE and STX 118) were found to have IC(50) values of 25-59 nM for the inhibition of hCAII activity. These compounds therefore have a similar CAII inhibitory potency to that of acetazolamide (IC(50)=25 nM), a known hCAII inhibitor. Docking studies have been performed with selected compounds to the crystal structure of hCAII and excellent correlation of scores with biological activity was observed. This agrees with our recent observations when we were the first to report the inhibition of hCAII by STS inhibitors. These studies and initial results with docking to the crystal structure of the extracellular domain of hCAXII indicate that the STS sulphamate ester inhibitors should also be interesting candidates to pursue as inhibitors of CA isozymes that are over-expressed in human tumours. PMID- 12767918 TI - The differentially expressed C21orf5 gene in the medial temporal-lobe system could play a role in mental retardation in Down syndrome and transgenic mice. AB - Mental retardation represents the more invalidating pathological aspect of Down syndrome, DS, and has a hard impact in public health. Modifications in DS brain, concerning abnormal size, neuronal differentiation, and cell density, cause changes in the neurophysiology and behavior of DS patients, and could be determined by dosage imbalance of genes localized in the DS critical region, DCR. Among these genes, C21orf5 showed high homology with Caenorhabditis elegans Pad1 involved in cellular differentiation and patterning. To shed light on C21orf5 role in DS, we performed molecular characterization of human and mouse orthologs, their spatio-temporal expression during development and in adult, and overexpression in DS and transgenic mice. C21orf5 was widely expressed early in embryogenesis in the nervous system. Later, its expression became differential and increased in mesencephalon and rhomboencephalon. This developmental expression profile evolves selectively in adult brain with higher signals in hippocampus, cerebellum, perirhinal, and entorhinal cortex, compared to the other cortical regions. Cellular specificity was detected in hippocampus with higher C21orf5 mRNA level in CA3 cells. Our findings appoint C21orf5 as candidate gene for mental retardation: Its overexpression in DS cells may contribute to gene imbalance in DS.Its specific expression in normal and its mirroring pattern in transgenic mice correspond to abnormal regions in DS patients and to neurological phenotype of transgenic mice. Altered cortical lamination in transgenic mice and the Pad1 ortholog function suggest a potential role of C21orf5 in cell differentiation. Its patterned differential expression in the medial temporal lobe system, including hippocampal formation and perirhinal cortex involved in memory storage, and learning and memory defects in the transgenic mice suggest a specialized role for C21orf5 in cognitive processes. These evidences suggest that C21orf5 is an attractive candidate gene contributing to neurological alterations responsible for mental retardation in DS patients. PMID- 12767919 TI - A novel murine long-chain acyl-CoA synthetase expressed in brain participates in neuronal cell proliferation. AB - Refsum disease (RfD) is an autosomal recessive neurologic disorder of the lipid metabolism. We have identified a novel murine long-chain acyl-CoA synthetase (mLACS) associated with the RfD gene using yeast two-hybrid assay. Northern blot analyses revealed that mLACS was expressed mainly in the brain and testis. mLACS was highly expressed in the brain at 2 weeks after birth and maintained through adult life. Expressions of the brain-specific LACS family increased in the PC12 cells undergoing neurite outgrowth by nerve growth factor. mLACS preferentially catalyzed the formation of arachidonoyl-CoA more than palmitoyl-CoA or oleoyl-CoA in PC12 cells. Triacsin C, an inhibitor of LACS, suppressed the cell proliferation and decreased mLACS expression in parent PC12 cells, but not in stably anti-sense mLACS cDNA-transfected cells. Our results indicate that mLACS participates in neuronal cell proliferation and differentiation, and interaction of the RfD gene with brain-selective mLACS may be involved in the pathogenesis of RfD. PMID- 12767920 TI - The ras-binding domain of ral GDS-like protein-2 as a ras inhibitor in smooth muscle cells. AB - This study was undertaken to determine whether the response of smooth muscle cells to mitogens can be inhibited by inactivating ras with the ral GDS like protein-2 ras-binding domain (RGL2-RBD). RGL2 is a member of the ral GDS family of proteins that contains a carboxy terminal ras-binding domain which binds the GTP ligated form of ras and rap and a CDC25 homology domain with the structural features of a guanine nucleotide exchange factor. The effect of ras signaling on the smooth muscle cell growth factor response was studied using rat aortic A10 smooth muscle cells transfected with a plasmid that encoded the RGL2-RBD. RGL2 RBD transfection resulted in a 12-fold reduction in the number of clonal colonies that were obtained after selection, and dramatically slowed cell cycle progression. RGBL2-RBD reduced DNA synthesis and inhibited platelet derived growth factor (PDGF)-mediated activation of the MAPK pathway. These findings indicated that interfering with ras signaling inhibits smooth muscle cell proliferation and raise the possibility that ras signaling inhibition might be used therapeutically to control smooth muscle proliferation after vascular injury. PMID- 12767921 TI - Direct observation of substrate induction of resistance mechanism in Pseudomonas aeruginosa using single live cell imaging. AB - The resistance mechanism in three strains of Pseudomonas aeruginosa, Delta ABM (devoid of MexAB-OprM), WT, and nalB-1 (overexpression of MexAB-OprM), was investigated using real-time single live cell imaging and fluorescence spectroscopy. Time courses of fluorescence intensity of these three strains in ethidium bromide (EtBr) showed that accumulation kinetics and extrusion machinery were highly dependent upon pump substrate (EtBr) concentration. At high substrate concentration (100 microM), the accumulation kinetic profiles in the cells at earlier incubation times were similar to those observed in low concentration. As EtBr accumulated in the cells reached a critical concentration, the fluorescence intensity of Delta ABM decreased below the fluorescence intensity of EtBr in buffer solution. This result suggested an inductive mechanism in the development of substrate resistance in P. aeruginosa. Substrates appeared to trigger the degradation of EtBr in Delta ABM. Unlike bulk measurements, single live cell imaging overcame the ensemble measurement of bulk analysis and showed that efflux machinery and resistance mechanism in individual cells were not synchronized. PMID- 12767922 TI - P21 response to DNA damage induced by genistein and etoposide in human lung cancer cells. AB - The p21(WAF1/Cip1) gene plays a central role in cell cycle regulation. Here we show that topoisomerase II inhibitors, genistein and etoposide, induce p21(WAF1/Cip1) expression mainly in a p53-dependent manner in human lung cancer cell line A549. However, although p53 accumulated, p21(WAF1/Cip1) expression did not depend on the level of Ser15 phosphorylation of p53. Caffeine, an ataxia telangiectasia-mutated (ATM), and ATM- and Rad3-related kinase (ATR) inhibitor, abrogated genistein-induced p21(WAF1/Cip1) and largely blocked etoposide-induced p21(WAF1/Cip1) expression. Wortmannin, an ATM- and DNA-dependent protein kinase inhibitor, partially inhibited p21(WAF1/Cip1) expression induced by genistein and etoposide, whereas UCN-01, a Chk1 inhibitor, partially blocked etoposide, but not genistein-induced p21(WAF1/Cip1) expression. These data suggest that both genistein and etoposide induce p21(WAF1/Cip1) expression in a p53-dependent manner. Genistein appears to stimulate p21(WAF1/Cip1) expression through p53 via ATM, whereas etoposide may activate both ATM and ATR pathways. Our results suggest different mechanisms participate in genistein and etoposide induced p21(WAF1/Cip1) expression. PMID- 12767923 TI - An apparent decrease in cholesterol biosynthesis in peroxisomal-defective Chinese hamster ovary cells is related to impaired mitochondrial oxidation. AB - Recent data suggest that impaired mitochondrial activities in Zellweger fibroblasts are related to defective peroxisome biogenesis and vice versa. To investigate the contribution of functional mitochondria to cholesterol biosynthesis, radioactive precursor molecules that form acetyl-CoA via beta oxidation-independent (pyruvate) or -dependent (palmitate and octanoate) pathways were used. Production of both 14C-labeled cholesterol and 14C-labeled CO(2) from these radioactive tracers was significantly impaired in peroxisomal-defective ZR 82 Chinese hamster ovary cells in comparison to controls. In contrast, cholesterol synthesis from acetate--a tracer directly converted to acetyl-CoA without the involvement of mitochondrial activities--was threefold higher in ZR 82 cells than in controls. Pathways further contributing to cellular cholesterol homeostasis, i.e., receptor-mediated binding of exogenous lipoprotein-associated cholesterol as well as intracellular mobilization of cholesteryl ester deposits were similar in ZR-82 and controls. From these findings, we propose that peroxisomal dysfunction in ZR-82 cells is tightly coupled to impaired mitochondrial activities, e.g., defective mitochondrial beta-oxidation and formation of acetyl-CoA from short chain fatty acids resulting in a decreased rate of CO(2) production, and an apparent decrease in cholesterol biosynthesis. Actually, cholesterol biosynthesis from acetate is increased in the peroxisomal defective cells. This explains previous conflicting conclusions. PMID- 12767924 TI - Odorant response assays for a heterologously expressed olfactory receptor. AB - Odorant responsiveness of a mouse olfactory receptor, mOR-EG, was investigated in various heterologous cells using a variety of detection methods. Odorant-induced Ca(2+) response was observed in HEK293 cells that coexpressed mOR-EG and the promiscuous G protein, G alpha 15. Without G alpha 15, a robust increase in cAMP level was observed upon odorant-stimulation in various mammalian cells. A luciferase reporter gene assay using zif268 promoter was adopted to amplify the cAMP signals. In Xenopus laevis oocytes, odorant-stimulated currents were recorded when mOR-EG cRNA was co-injected with either G alpha 15 or cAMP dependent channel. These results suggest that odorant responsiveness can be monitored via a signaling pathway mediated by endogenous G alphas or transfected G alpha 15 in heterologous cell systems. Various functional assays for a heterologously expressed olfactory receptor reported in this study, are potentially useful for high-throughput ligand screening and functional analyses of hundreds of olfactory receptors. PMID- 12767925 TI - DNA microarray analysis of human gingival fibroblasts from healthy and inflammatory gingival tissues. AB - In the inflammatory gingival tissues of patients with periodontitis, cytokines such as interleukin (IL)-1 alpha, IL-1 beta, IL-6, IL-8, and tumor necrosis factor (TNF)-alpha have been detected. Gingival fibroblasts are the major constituents of gingival tissue. We recently demonstrated that lipopolysaccharide (LPS) from periodontopathic bacteria induces inflammatory reactions in various tissues via CD14 and/or Toll-like receptors (TLRs) in gingival tissues [Biochem. Biophys. Res. Commun. 273 (2000) 1161]. To confirm this, we examined the expression of IL-1 alpha, IL-1 beta, IL-6, IL-8, TNF-alpha, CD14, TLR2, and TLR4 in human gingival fibroblasts (HGFs) obtained from patients with healthy or inflammatory gingiva using DNA microarray analysis. We also studied the expression levels of these proteins by flow cytometric analysis (FACS). The expression levels of all eight genes in the HGFs of the Inflammatory group were significantly higher than those in the Healthy group on DNA microarray analysis. FACS revealed that the expression levels of all eight proteins on the HGFs of the Inflammatory group were higher than those on the Healthy group. Our data indicated that these eight proteins in HGFs are involved in inflammatory conditions in the gingiva, including periodontal disease. Our results suggested that these eight proteins, in turn, act directly or indirectly on the immune response by activating host cells involved in inflammatory processes. PMID- 12767927 TI - Synleurin, a novel leucine-rich repeat protein that increases the intensity of pleiotropic cytokine responses. AB - We have identified and characterized a novel single span transmembrane leucine rich repeat protein, synleurin, that renders cells highly sensitive to the activation by cytokines and lipopolysaccharide (LPS). The major part of the extracellular domain consists of a leucine-rich repeats (LRR) cassette. The LRR central core has 12 analogous LRR repeating modules arranged in a seamless tandem array. The LRRs are most homologous to that of chondroadherin, insulin-like growth factor binding proteins, platelet glycoprotein V, slits, and toll-like receptors. Synleurin expression was detected at low levels in many tissues, including smooth muscle, brain, uterus, pancreas, cartilage, adipose, spleen, and testis. When synleurin is ecotopically expressed in transfected cells, the cells exhibit amplified responses to bFGF, EGF, PDGF-B, IGF-1, IGF-2, and LPS. Synleurin gene (slrn) maps to human chromosome at 5q12. The name synleurin reflects its synergistic effect on cytokine stimulation and its prominent leucine rich repeats. PMID- 12767926 TI - Cdk2 activity is associated with depolarization of mitochondrial membrane potential during apoptosis. AB - In this study we show that panaxadiol, a ginseng saponin with a dammarane skeleton, induces apoptotic cell death by depolarization of mitochondrial membrane potential in human hepatoma SK-HEP-1 cells. Sequential activation of caspases-9, -3, and -7, but not of caspase-8, occurs after mitochondrial membrane depolarization and cytochrome c release from the mitochondria of panaxadiol treated cells. Moreover, Cdk2 kinase activity, but not Cdc2 kinase activity, is markedly upregulated in the early stages of apoptosis. Olomoucine or roscovitine, specific Cdks inhibitors, effectively prevent mitochondrial membrane depolarization as well as apoptotic cell death in panaxadiol-treated cells. Thus, panaxadiol-treatment induces cell death-dependent activation of Cdk2 kinase activity, which is functionally associated with depolarization of mitochondrial membrane potential and subsequent cytochrome c release. PMID- 12767928 TI - Identification of the protein-protein contact site and interaction mode of human VDAC1 with Bcl-2 family proteins. AB - Bcl-2 family of proteins plays differential roles in regulation of mitochondria mediated apoptosis, by either promoting or inhibiting the release of apoptogenic molecules from mitochondria to cytosol. Bcl-2 family proteins modulate the mitochondrial permeability through interaction with adenine nucleotide translocator (ANT), voltage-dependent anion channel (VDAC), ADP/ATP exchange, or oxidative phosphorylation during apoptosis. Although the mitochondrial homeostasis is affected by the relative ratio of pro- and anti-apoptotic Bcl-2 family members, the molecular mechanism underlying the release of mitochondrial intermembrane proteins remains elusive. Here we reported the biochemical evidence that both pro-apoptotic Bax and anti-apoptotic Bcl-X(L) might simultaneously contact the putative loop regions of human VDAC1, and the existence of VDAC1-Bax Bcl-X(L) tertiary complex in vitro suggested that VDAC1 channel conformation and mitochondrial permeability could be determined by the delicate balance between Bax and Bcl-X(L). PMID- 12767929 TI - Crystallization and characterization of polyphosphate kinase from Escherichia coli. AB - Linear polyphosphate chains have been found to play a key role in bacterial responses to stresses and nutritional depletion, and are necessary for host infection of various pathogens. Polyphosphate kinase (PPK) is a critical enzyme responsible for polyphosphate synthesis in bacteria. PPK knockout mutations in several Gram-negative pathogens identify PPK as an ideal drug target for the development of a new class of antibacterial drugs. To reveal the catalytic mechanism and provide a structural basis for drug discovery, we have purified and crystallized full-length Escherichia coli PPK and its complex with AMP-PNP. The crystals diffract to a resolution of 2.5A and belong to the space group P4(2)2(1)2 with unit-cell parameters a=152.0, b=152.0, and c=150.0 A. Crystal structure of PPK is being determined by the Se-Met MAD experiment. PMID- 12767930 TI - Mice lacking Smad3 are protected against streptozotocin-induced diabetic glomerulopathy. AB - Transforming growth factor-beta (TGF-beta) has been implicated in the development of diabetic glomerulopathy. In order to evaluate a role of Smad3, one of the major signaling molecules downstream of TGF-beta, in the pathogenesis of diabetic glomerulopathy, Smad3-null mice were made diabetic with streptozotocin injection and analyzed 4 weeks after induction of diabetes. Electron microscopy revealed that the thickness of glomerular basement membrane (GBM) in wild-type diabetic mice was significantly higher than that in non-diabetic mice, whereas no appreciable GBM thickening was found in Smad3-null diabetic mice. Urinary albumin excretion was dramatically increased in wild-type diabetic mice, whereas Smad3 null diabetic mice did not show any overt albuminuria. Northern blotting revealed that mRNA levels of fibronectin and alpha 3 chain of type IV collagen (alpha 3Col4) in renal cortex of wild-type diabetic mice were approximately twice as much as those of non-diabetic mice, whereas their mRNA levels were not increased in Smad3-null diabetic mice. Real-time polymerase chain reaction (PCR) also confirmed diabetes-induced upregulation of fibronectin and alpha 3Col4 in glomeruli of wild-type mice. Glomerular expression of TGF-beta 1, as assessed by real-time PCR, was enhanced to a similar degree in wild-type and smad3-null diabetic mice, indicating that the observed differences between wild-type and Smad3-null mice are not attributable to difference in the expression of TGF-beta 1. These data clearly demonstrate a critical role of Smad3 in the early phase of diabetic glomerulopathy. This may be due at least partly to the present findings that diabetes-induced upregulation of fibronectin and alpha 3Col4 is dependent on Smad3 function. PMID- 12767931 TI - BRCA1 associates with human papillomavirus type 18 E2 and stimulates E2-dependent transcription. AB - BRCA1 is a breast and ovarian cancer-related tumor suppressor and has a role in transcriptional activation. We show here that BRCA1 enhances human papillomavirus type 18 (HPV-18) E2-dependent transcription in vivo. Using biochemical approaches, we discovered that BRCA1 interacts with the carboxyl-terminus of HPV 18 E2 protein in vivo and in vitro. Point mutations in the C-terminus make BRCA1 defective in transcriptional activation in E2-dependent transcription. This finding suggests that the C-terminus of BRCA1 is important for E2-dependent transcription. A chromatin immunoprecipitation assay indicated that BRCA1 is recruited onto the E2-dependent promoter region though E2 tethered to E2 binding sites in vivo. These results implicate that BRCA1 plays a role in E2-dependent transcription. PMID- 12767932 TI - Hyaluronan content in experimental carcinoma is not correlated to interstitial fluid pressure. AB - Mechanism(s) for generation of the high tumor interstitial fluid pressure (TIFP) that is characteristic of carcinoma is not known. We investigated the role of hyaluronan, the major water-binding polysaccharide of the extracellular matrix, for the generation of a high TIFP. A human anaplastic thyroid carcinoma (KAT-4) xenografted to athymic mice and a syngeneic rat colon carcinoma (PROb) were used. Neither KAT-4 nor PROb cells produced hyaluronan (HA) in culture, however, both cell lines produced factors that stimulated HA-synthesis by cultured fibroblasts. Modulating hyaluronan levels by transfection of PROb carcinoma cells with hyaluronan synthase-2 revealed no correlation between hyaluronan content and TIFP. Furthermore, lowering of TIFP by treating KAT-4 tumors with a specific inhibitor of TGF-beta 1 and -beta 3 did not change the concentration of hyaluronan in the tumors. In summary, our results suggest that a modulation of hyaluronan content is not a major pathogenetic mechanism for the generation of the characteristically high TIFP in malignant carcinomas. PMID- 12767933 TI - Permeability and gating properties of human connexins 26 and 30 expressed in HeLa cells. AB - Human connexins 26 and 30 were expressed either through the bicistronic pIRES EGFP expression vector or as EYFP-tagged chimeras. When transiently transfected in communication-incompetent HeLa cells, hCx26-pIRES transfectants were permeable to dyes up to 622 Da, but were significantly less permeable to 759 Da molecules. Under the same conditions, permeability of hCx26-EYFP fusion products was comparable to that of hCx26-pIRES, but with significant increase in diffusion at 759 Da, possibly as a consequence of having selected large fluorescent junctional plaques. Dye transfer was limited to 457 Da in hCx30-EYFP transfectants. When reconstructed from confocal serial sections, fluorescent plaques formed by hCx26 EYFP and hCx30-EYFP appeared irregular, often with long protrusions or deep invagination. Similar plaques were observed following immunostaining both in cells transfected with hCx26-pIRES and in HeLa cells stably transfected with mouse Cx26. Tissue conductance (Tg(j)) displayed significantly smaller values (28.8+/-1.8 nS) for stably transfected mCx26 than transiently transfected hCx26 (43.5+/-3.3 nS). These differences reflected in distinct functional dependence of normalized junctional conductance (G(j)) on transjunctional voltage (V(j)). The half-activation voltage for G(j) was close to +/-95 and +/-58 mV in mCx26 and hCx26, respectively. The corresponding parameters for hCx30 transfectants were Tg(j)= 45.2 +/- 3.5 nS and V(0)= +/- 34 mV. These results highlight unexpected differences between mCx26 and hCx26 in this expression system, reinforce the concept that channel permeability may be related to Cx level expression, and indicate that fusion of hCx30 to GFP colour mutants produces channels that are suitable for permeability and gating studies. PMID- 12767934 TI - Stimulation of cellular prion protein expression by TSH in human thyrocytes. AB - The cellular isoform of prion protein (PrP(C)) is a cell-surface glycosyl phosphatidylinositol-anchored protein which is ubiquitously expressed on the cell membrane. It may function as a cell receptor or as a cell adhesion molecule. Thyroid follicles, obtained from patients with Graves' disease at thyroidectomy, were cultured in F-12/RPMI-1640 medium supplemented with 0.5% fetal bovine serum and bovine thyroid stimulating hormone (bTSH). Northern blot analyses revealed that bTSH increased the steady-state expression levels of PrP mRNA in a time- and dose-dependent manner. This increase was reproduced by dibutyryl-cAMP and 12 decanoylphorbol-13-acetate. The mRNA expression was greater in thyroid follicles in suspension culture than in thyrocytes cultured in a monolayer. These findings suggest that TSH stimulates PrP mRNA expression in thyrocytes through the protein kinase A and C pathways. The greater mRNA expression in thyroid follicles than in monolayer cells suggests that PrP(C) may be involved in structure formation or maintenance of thyroid follicles. PMID- 12767935 TI - Sonic hedgehog and FGF8 collaborate to induce dopaminergic phenotypes in the Nurr1-overexpressing neural stem cell. AB - Neural stem cells are self-renewing cells capable of differentiating into all neural lineage cells in vivo and in vitro. In the present study, coordinated induction of midbrain dopaminergic phenotypes in an immortalized multipotent neural stem cell line can be achieved by both overexpression of nuclear receptor Nurr1, and fibroblast growth factor-8 (FGF-8), and sonic hedgehog (Shh) signals. Nurr1 overexpression induces neuronal differentiation and confers competence to respond to extrinsic signals such as Shh and FGF-8 that induce dopaminergic fate in a mouse neural stem cell line. Our findings suggest that immortalized NSCs can serve as an excellent model for understanding mechanisms that regulate specification of ventral midbrain DA neurons and as an unlimited source of DA progenitors for treating Parkinson disease patients by cell replacement. PMID- 12767936 TI - Association of hepatitis B virus polymerase with promyelocytic leukemia nuclear bodies mediated by the S100 family protein p11. AB - Hepatitis B virus (HBV) polymerase (Pol) interacts with cellular chaperone proteins and thereby performs multiple functions necessary for viral replication. Yeast two-hybrid analysis was applied to identify additional cellular targets required for HBV Pol function. HBV Pol interacted with S100A10 (p11), a Ca(2+) modulated protein previously shown to bind to annexin II. The interaction between HBV Pol and p11 was confirmed by co-immunoprecipitation of the two proteins synthesized either in vitro or in transfected cells and by inhibition of the DNA polymerase activity of HBV Pol by p11. Immunofluorescence analysis of transfected human cell lines revealed that, although most HBV Pol and p11 was restricted to the cytoplasm, a small proportion of each protein colocalized as nuclear speckles; HBV Pol was not detected in the nucleus in the absence of p11. The HBV Pol-p11 nuclear speckles coincided with nuclear bodies containing the promyelocytic leukemia protein PML. Furthermore, the association of HBV Pol-p11 with PML was increased by exposure of cells to EGTA and inhibited by valinomycin. These results suggest a role for p11 in modulation of HBV Pol function and implicate PML nuclear bodies and intracellular Ca(2+) in viral replication. PMID- 12767937 TI - Role of Shiga toxin 2 (Stx2)-binding protein, human serum amyloid P component (HuSAP), in Shiga toxin-producing Escherichia coli infections: assumption from in vitro and in vivo study using HuSAP and anti-Stx2 humanized monoclonal antibody TMA-15. AB - Shiga toxin 2 (Stx2) is a major pathogenic factor in Shiga toxin-producing Escherichia coli (STEC) infections. Some factor that neutralizes Stx2 in vitro had been shown to be specifically present in human serum and we recently identified it as human serum amyloid P component (HuSAP). Here, we report the role of HuSAP in STEC infections. HuSAP could not rescue Stx2-challenged mice from death, and it instead reduced the efficacy of the Stx2-neutralizing humanized monoclonal antibody TMA-15 when a lower dose of TMA-15 was injected to the mice. By contrast, the efficacy of TMA-15 at a higher dose was uninfluenced by the presence of HuSAP. These findings suggest that HuSAP acts as a carrier protein of Stx2 rather than as a Stx2-neutralizing factor in the human circulation and that passive immune therapy with Stx2-neutralizing antibodies such as TMA-15 is useful to prevent severe complications associated with STEC infections even in the presence of HuSAP. PMID- 12767938 TI - Selection of peptides with affinity for the N-terminal domain of GATA-1: Identification of a potential interacting protein. AB - As most transcription factors, GATA-1 activities are mediated by interactions with multiple proteins. Those identified so far associate with the zinc-finger domain and/or surrounding sequences. In contrast, no proteins interacting with the N-terminal domain have been identified although several evidences suggest its involvement in the control of hematopoiesis. In an attempt to identify proteins that interact with the N-terminal transactivation domain of GATA-1, a random phage peptide library was screened with recombinant GATA-1 protein and the sequence of a selected peptide was used for database protein sequence retrieval. We selected a set of peptides sharing the core sequence phi-B((2-3))-nu((2-4)) (where phi, B, and nu represent hydrophobic, basic, and neutral residues, respectively). Using the sequence of the most represented peptide (pep5) as query, we retrieved the HIV accessory protein Nef. We show that Nef binds GATA-1 and GATA-3 in vitro in virtue of its sequence homology with pep5. PMID- 12767939 TI - Mutational and sequence analysis of transmembrane segment 6 orientation in TetA proteins. AB - The packing orientations of the 8 transmembrane (TM) segments that line the central, aqueous transport channel within tetracycline resistance proteins (TetA) have been established. However, the orientations of the remaining 4 segments, TMs 3, 6, 9, and 12, located at the periphery, and away from the transport channel, have not yet been determined. In this study, the packing orientation of TM6 within the class C TetA protein encoded by plasmid pBR322 was evaluated by substitution mutagenesis and analysis of sequence conservation and amphipathicity. The combined data support a model in which the conserved and polar face of the TM6 alpha-helix containing Asn170 and Asn173 orients towards channel-lining TM segments, and the relatively non-conserved and hydrophobic face of TM6 points towards membrane lipids. PMID- 12767940 TI - Mouse MTH2 protein which prevents mutations caused by 8-oxoguanine nucleotides. AB - MutT-related proteins degrade 8-oxo-7,8-dihydrodeoxyguanosine triphosphate (8-oxo dGTP), a mutagenic substrate for DNA synthesis, in the nucleotide pool, thereby preventing DNA replication errors. During a search of GenBank EST database, we found a new member of MutT-related protein, MTH2, which possesses the 23-amino acid MutT module. The cloned mouse MTH2 (mMTH2) cDNA was expressed in Escherichia coli mutT(-) cells and the protein was purified. mMTH2 protein hydrolyzes 8-oxo dGTP to 8-oxo-dGMP, with Km of 32 microM. Expression of cDNA for mMTH2 reduced significantly the elevated level of spontaneous mutation frequency of E. coli mutT(-) cells. Thus, MTH2 has a potential to protect the genetic material from the untoward effects of endogenous oxygen radicals. MTH2 could act as an MTH1 redundancy factor. PMID- 12767941 TI - Role of context in the relationship between form and function: structural plasticity of some PROSITE patterns. AB - True positive hits of PROSITE sequence pattern are expected to have a characteristic three-dimensional structure. The combined sequence-structure attributes of PROSITE patterns can be used for function prediction of an uncharacterized protein with known primary and 3D structure, a situation that might arise in structural genomics projects. We have found specific examples of true hits of PROSITE patterns displaying structural plasticity by assuming significantly different local conformation, depending upon the context. Our work highlights the importance of taking into account all the known distinct conformations of PROSITE patterns, while creating a sensitive 3D template for the pattern, for use in functional annotation. PMID- 12767942 TI - Ultrastructural evidences of HCV infection in hepatocytes of chronically HCV infected patients. AB - In this study, 13 samples of liver biopsies from patients with chronic hepatitis C were studied by transmission electron microscopy (EM) and immunoelectron microscopy (IEM). The 13 biopsies showed ultrastructural cell damage typical of acute viral hepatitis. In four of the 13 liver biopsies enveloped virus-like particles (VLPs) inside cytoplasmic vesicles and in the cytoplasm of hepatocytes were observed. We also detected the presence of unenveloped VLPs mainly in the cytoplasm and in the endoplasmic reticulum. IEM using anti-core, E1 and E2 monoclonal antibodies (mAbs) confirmed the specific localization of these proteins, in vivo, inside cytoplasm and endoplasmic reticulum. Thus, this work provided evidence for hepatocellular injury related to HCV infection. It also suggested the presence of HCV-related replicating structures in the cytoplasm of hepatocytes and raised the possibility of hepatitis C virion morphogenesis in intracellular vesicles. PMID- 12767943 TI - Citraconylation--a simple method for high protein sequence coverage in MALDI-TOF mass spectrometry. AB - Lysine epsilon -amino group reacts with citraconic anhydride forming a derivative, which is stable on terms for trypsin cleavage. This modification changes the spectrum of peptides formed by the trypsin action; as the number of trypsin-sensitive sites is reduced, the peptides with higher molecular mass can survive in the digest. The various studies of proteins by MALDI-TOF mass spectrometry are often complicated by the low sequence coverage of the peptide chain. This paper demonstrates that the modification of proteins by citraconylation before trypsin cleavage represents a simple experimental technique, which allows a significant increase of sequence coverage in MALDI-TOF mass spectrometry. This improvement is caused both by change of trypsin fragmentation pattern and by disturbance of the protein's native tertiary structure. PMID- 12767944 TI - Phosphorylation of NF-kappa B by calmodulin-dependent kinase IV activates anti apoptotic gene expression. AB - We previously presented that calmodulin-dependent kinase IV (CaMKIV) mutually interacts with NF-kappa B and phosphorylates it directly, inducing the increased transcriptional regulation dependent on NF-kappa B target genes [J. Biol. Chem. 276 (2001) 20005]. Here, we show that Ser(535) residue is phosphorylated by CaMKIV. S535A mutant of p65 was specifically defective in transactivation of NF kappa B target gene expression induced by CaMKIV. While coexpression of active CaMKIV with wild-type p65 led to a recovery from etoposide-induced apoptosis and an increase of Bcl-2 protein in cells, cells expressing S535A mutant did not. Taken together these results suggest that phosphorylated NF-kappa B p65 on Ser(535) by CaMKIV increases NF-kappa B target gene expression, including anti apoptotic gene, hence leading to inhibition of apoptosis. PMID- 12767945 TI - The adenoviral E1A induces p21WAF1/CIP1 expression in cancer cells. AB - The adenovirus-5 E1A gene encodes two main proteins of 289 and 243 amino acid residues from 13S and 12S mRNA, respectively. The E1A gene products function as transcriptional regulators and have anti-tumor activities. Despite the fact that E1A gene therapy has been tested in clinical trials, the molecular mechanism by which it suppresses tumor cell growth is still not completely understood. Here, we show that E1A increases the expression of the cyclin-dependent kinase (CDK) inhibitor p21(WAF1/CIP1), which inhibits cell growth. We further show that 13S E1A, but not 12S E1A, can transactivate the p21 promoter through Sp1 sites. Interestingly, the E1A-induced transactivation occurs only in cancer cells, not in normal cells. This study provides new insight into the links between E1A and the CDK inhibitor and may have important clinical implications. PMID- 12767946 TI - The CAG repeats in exon 1 of the androgen receptor gene are significantly longer in endometrial cancer patients. AB - Human androgen receptor (AR) gene contains polymorphic CAG repeats in the N terminal domain that influence transcription efficiency. The CAG repeats encode a poly-glutamine tract, which has a length that is inversely and linearly related to AR activity. It has been observed that longer CAG repeats impose a lower transactivation activity on AR and have a decreased binding affinity for androgens. Androgens have an anti-proliferative effect on endometrial cells. We hypothesize that the length of CAG repeats on the AR gene can predict higher incidence of endometrial cancer. To test this hypothesis, the genetic distributions of CAG repeats on AR gene polymorphisms were investigated in endometrial cancer patients and healthy controls. Genotyping and a chi(2) test revealed that the distribution of CAG repeats was significantly different between the endometrial cancer patients and normal healthy controls (P<0.001). The endometrial cancer patients had longer alleles than normal healthy controls. The longer CAG repeats in the AR gene may cause a decrease of transactivation function in the receptor, weaken an anti-proliferative effect on uterine endometrial cells, and promote carcinogenesis of the uterine endometrial cells. The findings suggest that the CAG repeats in the AR gene may be important in the carcinogenesis of uterine endometrial cells. PMID- 12767947 TI - Differential UVC-induced gadd45 gene expression in xeroderma pigmentosum cells. AB - Xeroderma pigmentosum C (XPC) is a DNA repair factor essential for global genome repair (GGR) in nucleotide excision repair (NER). In the present study we screened for factors regulated by XPC after DNA damage. Ultraviolet C (UVC) irradiation-induced stress response factors were analyzed by a cDNA microarray chip system in HeLa and XP4PA-SV xpc mutant cell lines. The principal component analysis (PCA) method was employed to identify groups of genes with similar expression patterns over time after UVC irradiation. The growth arrest and DNA damage-inducible gene gadd45, as well as a small group of other genes, was found to exhibit an inducible expression pattern after 30min of incubation in xpc mutants but not in HeLa cells. Subsequent studies showed that gadd45 gene expression post-UVC irradiation was also present in the GGR mutant cells xpa and xpd, but not in TCR mutant csb cells. This evidence indicates that gadd45 plays a regulatory role in GGR of NER. PMID- 12767948 TI - Two collaborating artists produce a work of art: the medical interview. PMID- 12767949 TI - "Building" a history rather than "taking" one: a perspective on information sharing during the medical interview. AB - Patients and physicians enter the medical encounter with unique perspectives on the illness experience. These perspectives influence the way that information is shared during the initial phase of the interview. Previous research has demonstrated that patients who are able to fully share their perspective often achieve better outcomes. However, studies of patient-physician communication have shown that the patient's perspective is often lost. Researchers and educators have responded with calls for practitioners to adopt a "narrative-based medicine" approach to the medical interview. In this article, we review the literature on narrative-based medicine with an emphasis on information sharing during the medical interview. We suggest a framework of skills and attitudes that can act as a foundation for future work in educating practitioners and researching the medical interview. PMID- 12767950 TI - Aspirin and clopidogrel in acute coronary syndromes: therapeutic insights from the CURE study. AB - Platelet adhesion, activation, and aggregation are central to thrombus formation, which follows atherosclerotic plaque disruption and causes acute coronary syndromes. Aspirin and clopidogrel exert their antiplatelet effects by inhibiting thromboxane A2 production and adenosine diphosphate-induced platelet aggregation pathways, respectively. Aspirin has proven benefits in primary and secondary prevention of coronary artery disease. Clopidogrel, an alternative antiplatelet agent used in patients with aspirin intolerance, is especially useful in combination with aspirin after coronary stent procedures. The CURE (Clopidogrel in Unstable Angina to Prevent Recurrent Events) study demonstrates for the first time the benefit of adding clopidogrel to aspirin rather than using aspirin alone in patients having acute coronary syndromes without ST-segment elevation myocardial infarction. Patients who are resistant to aspirin (up to 10%) have higher rates of cardiovascular events and may derive special benefit from the combination therapy. Aspirin resistance can be assessed through platelet aggregometry testing, measurement of urinary thromboxane metabolites, and, possibly, genomic testing in the future. PMID- 12767951 TI - Renin-angiotensin system and atherothrombotic disease: from genes to treatment. AB - The renin-angiotensin system plays a central role in the pathogenesis of cardiovascular disease. At the molecular and cellular levels, angiotensin II, the main effector peptide of the system, stimulates key components of atherosclerosis. Trials in animals and humans indicate that blocking renin angiotensin system pathways decreases atherosclerotic plaque progression and ischemic events. This review provides a broad overview of the entire role of the renin-angiotensin system in atherothrombotic disease, ranging from molecular pathways to human genetics to the latest clinical trials. PMID- 12767952 TI - A randomized controlled trial of test-and-treat strategy for Helicobacter pylori: clinical outcomes and health care costs in a managed care population receiving long-term acid suppression therapy for physician-diagnosed peptic ulcer disease. AB - BACKGROUND: Guidelines recommend Helicobacter pylori (HP) testing and treatment for patients with a history of peptic ulcer disease (PUD), assuming that PUD has been documented and that successful HP eradication would eliminate the need for further therapy and medical utilization. METHODS: An open-label, randomized controlled trial in a managed care setting evaluated the clinical outcome and costs of an HP test-and-treat (T & T) strategy in 650 patients receiving long term acid suppression therapy for physician-diagnosed PUD. Patients were randomized to T & T for HP (n = 321) or to usual care (n = 329). Outcome measures included presence and severity of PUD symptoms, use of acid-reducing medication, and acid-peptic-related health care costs during 12-month follow-up. RESULTS: Only 17% of study participants had PUD confirmed by radiography or endoscopy; only 38% of the T & T group tested positive for HP. At 12 months, patients in the T & T group were less likely to report ulcerlike dyspepsia or use of acid reducing medication; however, 75% of the T & T group used acid-reducing medication during the second half of the 12-month follow-up. In the 12 months after randomization, the T & T group had higher total acid-peptic-related costs than the usual care group. CONCLUSIONS: Most patients receiving long-term acid suppression therapy for physician-diagnosed PUD in community practice settings are likely to have HP-negative, uninvestigated dyspepsia. Routine testing and treating for HP will not reduce acid-peptic-related costs and have only a modest (though statistically significant) effect in reducing clinical symptoms and use of acid-reducing medications. PMID- 12767953 TI - Relationship between periodontal disease and C-reactive protein among adults in the Atherosclerosis Risk in Communities study. AB - BACKGROUND: Moderately elevated serum C-reactive protein (CRP) concentration is a systemic marker of inflammation and a documented risk factor for cardiovascular disease in otherwise healthy persons. Unrecognized infections, such as periodontal disease, may induce an acute-phase response, elevating CRP levels. We evaluated the association between periodontal disease and CRP levels in adults in the Atherosclerosis Risk in Communities study. METHODS: Oral examinations were conducted between January 1, 1996, and December 31, 1998, on 5552 ARIC participants (aged 52-74 years) from 4 US communities. Periodontal disease was quantified as the percentage of periodontal sites with pocket depth of 4 mm or more. Serum CRP concentration was quantified in milligrams per liter using an enzyme-linked immunosorbent assay. RESULTS: Mean (SE) CRP level was 7.6 (0.6) mg/L among people with extensive periodontal pockets (>30% of sites with pocket depth > or =4 mm), approximately one-third greater than that for people with less extensive periodontal pockets (5.7 [0.1] mg/L). In a multivariable linear regression model that controlled for age, sex, diabetes mellitus, cigarette use, and nonsteroidal anti-inflammatory drug use, the association of extensive periodontal pockets with CRP concentration was modified by body mass index (BMI; calculated as weight in kilograms divided by the square of height in meters). For people with a BMI of 20, the model predicted a 2-fold difference in mean CRP concentration between periodontal pocket groups (7.5 vs 3.6 mg/L), but the difference decreased with increasing BMI and was negligible when BMI equaled 35. CONCLUSIONS: Extensive periodontal disease and BMI are jointly associated with increased CRP levels in otherwise healthy, middle-aged adults, suggesting the need for medical and dental diagnoses when evaluating sources of acute-phase response in some patients. PMID- 12767954 TI - In-hospital mortality following acute exacerbations of chronic obstructive pulmonary disease. AB - BACKGROUND: Acute exacerbations of chronic obstructive pulmonary disease (COPD) are a frequent cause of hospitalization in the United States. Previous studies of selected populations of patients with COPD have estimated in-hospital mortality to range from 4% to 30%. Our objective was to obtain a generalizable estimate of in-hospital mortality from acute exacerbation of COPD in the United States and to identify predictors of in-hospital mortality using administrative data. METHODS: We performed a cross-sectional study utilizing the 1996 Nationwide Inpatient Sample, a data set of all hospitalizations from a 20% sample of nonfederal US hospitals. The study population included 71 130 patients aged 40 years or older with an acute exacerbation of COPD at hospital discharge. The primary outcome assessed was in-hospital mortality. RESULTS: In-hospital mortality for patients with an acute exacerbation of COPD was 2.5%. Multivariable analyses identified older age, male sex, higher income, nonroutine admission sources, and more comorbid conditions as independent risk factors for in-hospital mortality. CONCLUSIONS: Mortality during hospitalization in this nationwide sample of patients with acute exacerbations of COPD was lower than that of previous studies of select populations. This estimate should provide optimism to both clinicians and patients regarding prognoses from COPD exacerbations requiring hospitalization. Our results indicate that the use of administrative data can help to identify subsets of patients with acute exacerbations of COPD that are at higher risk of in-hospital mortality. PMID- 12767955 TI - Alcohol consumption and mortality in men with preexisting cerebrovascular disease. AB - BACKGROUND: In counseling patients with a history of stroke, clinicians have limited information regarding the risks and benefits of alcohol consumption. OBJECTIVE: To examine the relationship between alcohol intake and risks of total and cardiovascular mortality in men with a history of stroke. METHODS: The study population consisted of 112 528 men from the enrollment cohort of the Physicians' Health Study, 1320 of whom reported a baseline history of stroke. Men provided self-reported data on alcohol consumption, which was classified into 1 of 4 categories: rarely or never drink, very light (<1 drink per week), light (1-6 drinks per week), or moderate (> or =1 drink per day). Cox proportional hazards models were used to assess the relative risks of mortality associated with alcohol consumption, after adjustment for major coronary risk factors. RESULTS: During a mean follow-up of 4(1/2) years, 369 men died, 267 of whom died of cardiovascular disease. Compared with men with a history of stroke who drank rarely or never, those with a very light to moderate alcohol intake had multivariate relative risks for total mortality of 0.88 (95% confidence interval [CI], 0.60-1.28), 0.64 (95% CI, 0.48-0.85), and 0.71 (95% CI, 0.54-0.94), respectively (P =.03 for trend); and relative risks for cardiovascular mortality of 0.89 (95% CI, 0.58-1.36), 0.56 (95% CI, 0.40-0.79), and 0.64 (95% CI, 0.46 0.88) P =.008 for trend). Compared with age-adjusted models, adjustment for major coronary risk factors did not significantly change risk estimates for total or cardiovascular mortality. CONCLUSIONS: These data indicate a possible inverse association between light to moderate alcohol intake and risks of total and cardiovascular mortality in men with a history of stroke. More data are needed to confirm or refute these results. PMID- 12767956 TI - The additional diagnostic value of ultrasonography in the diagnosis of breast cancer. AB - BACKGROUND: The use of ultrasonography (US) in diagnostic breast imaging is increasing. Restricting US to subgroups of patients who benefit most would result in a more efficient and effective application. This study assessed the diagnostic value of US as an adjunct to mammography (MAM) and a clinical examination (CE) in the diagnosis of breast cancer and the feasibility of selecting subgroups of patients who benefit the most. METHODS: Between October 1, 1999, and August 1, 2000, all consecutive patients referred for breast imaging underwent additional US after MAM and a CE. Results were scored on a 5-point grading scale of increasing suggestion of malignancy. Pathologic results during 12 months of follow-up were used as the criterion standard. Receiver-operating characteristic curve analysis assessed the diagnostic value of US in the whole population and in subgroups of patients according to indication for referral and age. RESULTS: A total of 3835 breasts were examined in 2020 patients, with a 6.3% prevalence of breast cancer. Breast US detected 8 extra malignancies and correctly downgraded 332 cases from a positive to a negative diagnosis (ie, from a suggested malignancy to no malignancy). Receiver-operating characteristic curves showed a significant improvement in diagnostic value by adding US to MAM and a CE (area under the curve for CE + MAM + US vs CE + MAM, 0.99 vs 0.95; P =.002). The diagnostic yield improved significantly in patients referred for palpable breast lumps (P =.004) or referred from the National Breast Cancer Screening Program (P =.05). Less pronounced was the value in patients referred for other symptoms or for follow-up of a prior breast malignancy. When breast imaging of the contralateral breast or of asymptomatic patients referred for reassurance or follow-up of a prior benign lesion was performed, the value of additional US remained undefined because of the few malignancies found. CONCLUSIONS: The systematic application of breast US improved the overall diagnostic yield. The diagnostic value increased most in patients with palpable breast lumps and in patients referred with abnormal screening MAM results. PMID- 12767957 TI - Physical activity, coronary heart disease, and inflammatory response. AB - BACKGROUND: We sought to estimate the risk for coronary heart disease (CHD) associated with leisure time physical activity (LTPA) and work-related physical strain (WRPS) after careful adjustment for other established risk factors and to elucidate the association of physical activity with various hemostatic and inflammatory markers. METHODS: Case-control study including 312 patients aged 40 to 68 years with stable CHD (angiographically confirmed) and 479 age- and sex matched controls. Main outcome measures were odds ratio for CHD associated with LTPA and WRPS and associations of physical activity with inflammatory and other biochemical markers after adjustment for covariates. RESULTS: LTPA showed a clear inverse association with risk of CHD. Compared with subjects who reported no summer LTPA, the odds ratio for CHD was 0.85 (95% confidence interval [CI], 0.47 1.53) in the category <1 h/wk; 0.60 (95% CI, 0.38-0.95) in the category 1-2 h/wk; and 0.39 (95% CI, 0.26-0.59) in the category >2 h/wk, after full adjustment for covariates. Similar results were obtained for winter LTPA. By contrast, there was a strong positive association between WRPS and risk of CHD. Furthermore, levels of C-reactive protein, serum amyloid A, interleukin 6, and intercellular adhesion molecule 1 were inversely and independently associated with LTPA, but not with WRPS. CONCLUSIONS: This study provides further evidence that LTPA, but not WRPS, is associated with a decreased risk of CHD, effective at even moderate levels. It further demonstrates that LTPA is associated with beneficial effects on the inflammatory response. This may represent one mechanism to explain the benefits of LTPA on coronary risk. PMID- 12767958 TI - Acute pyelonephritis in adults: prediction of mortality and failure of treatment. AB - BACKGROUND: To formulate a classification tool for early recognition of patients admitted with acute pyelonephritis (AP) who are at high risk for failure of treatment or for death. METHODS: A retrospective chart review of 225 patients (102 men) admitted with AP. We considered 13 potential risk factors in a multivariate analysis. RESULTS: Recent hospitalization, previous use of antibiotics, and immunosuppression were found to be independent correlates of the prevalence of resistant pathogens in both sexes. Additional predictors included nephrolithiasis in women and a history of recurrent AP in men. Prolonged hospitalization should be expected for a man with diabetes and long-term catheterization who is older than 65 years or for a woman of any age with the same characteristics, when the initial treatment was changed according to the results of urine culture. For mortality prediction, we derived an integer-based scoring system with 6 points for shock, 4 for bedridden status, 4 for age greater than 65 years, and 3 for previous antibiotic treatment for men and 6 points for shock, 4 for bedridden status, 4 for age greater than 65 years, and 3 for immunosuppression for women. Among patients with at least 11 points, the risk for in-hospital death was 100% for men and 91% for women. CONCLUSIONS: Simple variables available at presentation can be used for risk stratification of patients with AP. The additional identification of certain risk factors by means of a carefully obtained history could contribute to early recognition of patients infected by resistant bacteria and optimize the selection of antimicrobial agents. PMID- 12767959 TI - Tracking the uptake of evidence: two decades of hospital practice trends for diagnosing deep vein thrombosis and pulmonary embolism. AB - BACKGROUND: Advances in clinical research methods have led to prospective randomized controlled (level 1) clinical studies evaluating diagnostic modalities resulting in a paradigm shift in the literature for diagnosing deep vein thrombosis (DVT) and pulmonary embolism (PE). To assess whether these advances correlate with clinical practice, we analyzed 21-year trends in diagnostic testing for patients with venous thromboembolism. METHODS: We used discharge data from the National Hospital Discharge Survey (1979-1999) to determine DVT and PE cases annually. Procedure fields were screened to determine patients who had DVT or PE or who underwent venography, arteriography of the pulmonary arteries, pulmonary scintigraphy, or DVT ultrasonic scanning. Searching EMBASE, MEDLINE, and the American Thoracic Society guidelines, a literature-based time line of level 1 studies was derived and juxtaposed against trends and procedure use. RESULTS: Improved diagnostic tests resulted in diagnostic changes in patients with suspected venous thromboembolism. These observed changes correlated over time in subsequent years with level 1 studies. Diagnostic DVT approaches showed an initial marked increased use of venography followed by a rapid decline that coincided with increased use of Doppler ultrasonography. Diagnostic approaches to PE were characterized by initial marked increases in lung scanning followed by a rapid decline as use of ultrasonography considerably increased and pulmonary angiography modestly increased. CONCLUSIONS: Diagnostic approaches to DVT and PE have changed markedly during the past 2 decades, in temporal harmony with the evolving literature. Change in clinical practice occurs over years, and long-term follow-up is required to capture this change. PMID- 12767961 TI - Clinical studies of sudden upper airway obstruction in patients with hereditary angioedema due to C1 esterase inhibitor deficiency. AB - BACKGROUND: Hereditary angioedema due to C1 esterase inhibitor deficiency is clinically characterized by recurrent and self-limiting skin, intestinal, and laryngeal edema. Asphyxiation by laryngeal edema is the main cause of death among patients who die of hereditary angioedema. This study describes the age at which laryngeal edema first occurs, the time between onset and full development, and the effectiveness of therapy and prophylaxis. METHODS: Information on 123 patients with hereditary angioedema was obtained from medical histories and reports by the general practitioners, emergency physicians, and hospitals involved. RESULTS: Sixty-one patients (49.6%) experienced a total of 596 laryngeal edema episodes. The ratio of laryngeal edema episodes to skin swellings and abdominal pain attacks was approximately 1:70:54 in patients who had laryngeal edema. The mean (SD) age at the first laryngeal edema was 26.2 (15.3) years. Nearly 80% of the laryngeal edemas occurred between the ages of 11 and 45 years. The mean interval between onset and maximum development of laryngeal edema was 8.3 hours. A total of 342 laryngeal edemas cleared spontaneously without treatment, and 208 laryngeal edemas were successfully treated with C1 esterase inhibitor concentrate. Despite long-term prophylactic treatment with danazol, 6 patients developed subsequent laryngeal edemas. CONCLUSIONS: Laryngeal edema may occur at any age, although young adults are at greatest risk. In adults, the interval between onset of symptoms and acute risk of asphyxiation is usually long enough to allow for use of appropriate emergency procedures. To prevent a fatal outcome, it is essential to instruct patients and their relatives about the first signs of laryngeal edemas and the necessary procedures to follow. PMID- 12767960 TI - A prospective trial of structured treatment interruptions in human immunodeficiency virus infection. AB - BACKGROUND: According to the "autovaccination hypothesis," reexposure to human immunodeficiency virus (HIV) during treatment interruptions may stimulate the HIV specific immune response and lead to low viremia after withdrawal of highly active antiretroviral treatment (HAART). Many patients who started HAART earlier in their disease course than is currently recommended would like to discontinue, but it is unknown whether it is safe to do so. OBJECTIVES: To determine whether repeated treatment interruptions of HAART (1) stimulated the cytotoxic HIV specific immune response and whether such stimulation correlated with low viremia off treatment, and (2) were safe with respect to clinical complications, development of viral resistance, and decline in CD4 cell counts. DESIGN: Interventional study with before-after comparison. SETTING: Outpatient clinics of university hospitals in Switzerland and Spain. PATIENTS: A total of 133 patients receiving HAART, with a median CD4 cell count of 740/ microL, and whose viral load had been undetectable for a median of 21 months. INTERVENTIONS: HAART was interrupted for 2 weeks, restarted, and continued for 8 weeks. After 4 such cycles, treatment was indefinitely suspended 40 weeks after study entry. MAIN OUTCOME MEASURES: HIV-specific cytotoxic T-cell responses were evaluated by interferon gamma enzyme-linked immunospot analysis. The proportion of "responders" (viral load <5000 copies/mL) was measured at weeks 52 and 96. HIV related diseases and CD4 cell counts were recorded. RESULTS: Seventeen percent of patients (95% confidence interval, 11%-25%) were responders at week 52, and 8% at week 96. Low pre-HAART viral load and lack of rebound during weeks 0 to 40 predicted response. HIV-specific CD8+ T cells increased between week 0 (median, 343 spot-forming cells per million peripheral blood lymphocytes [SFC/106 PBL]) and week 52 (median, 1930 SFC/106 PBL), but there was an inverse correlation between response and the number of spot-forming cells. Eighty-five (64%) of 133 patients stopped therapy for at least 12 weeks, and 55 (41%) for at least 56 weeks. The median CD4 cell count decreased from 792/ microL to 615/ microL during the first 12 weeks without treatment, but stabilized thereafter. One patient (0.75%) developed drug resistance necessitating salvage treatment. There were no AIDS-related clinical complications. CONCLUSIONS: Results of this study do not favor the autovaccination hypothesis. Treatment interruptions did not provoke clinical complications, and there was little drug resistance. Comparative trials will have to show what benefit, if any, is associated with intermittent, as opposed to continuous treatment. PMID- 12767962 TI - Undertreatment of osteoporosis in men who have had a hip fracture. PMID- 12767963 TI - Hypogonadism and osteoporosis in men. PMID- 12767964 TI - Thrombolysis in the treatment of submassive pulmonary embolism. PMID- 12767965 TI - Cat-associated zoonosis: don't forget rabies and leishmaniasis. PMID- 12767966 TI - Pasteurella multocida respiratory infection: an important cat-associated zoonosis. PMID- 12767967 TI - Is orthostatic hypotension a consistent finding in the acute geriatric ward? PMID- 12767968 TI - Difference in breast cancer stage, treatment, and survival. PMID- 12767969 TI - New is not always better. PMID- 12767970 TI - Anticoagulated patients and oral surgery. PMID- 12767971 TI - International normalized ratio and anticoagulation. PMID- 12767972 TI - Apparent warfarin potency inconsistencies compromise effectiveness. PMID- 12767973 TI - Is low-molecular-weight heparin safer than warfarin for secondary prevention of venous thromboembolism in cancer patients? PMID- 12767974 TI - Lysosomal and proteasomal degradation play distinct roles in the life cycle of Cx43 in gap junctional intercellular communication-deficient and -competent breast tumor cells. AB - The present study was designed to determine the specific roles played by lysosomes and proteasomes in the degradation of Cx43 in both gap junctional intercellular communication-deficient MDA-MB-231 and -competent BICR-M1Rk cells. In MDA-MB-231 cells, immunolocalization and brefeldin A protein transport blocking studies revealed that there was a propensity for newly synthesized Cx43 to be transported to lysosomes. On the other hand, light and electron microscopic analysis of BICR-M1Rk cells showed that Cx43 gap junctions were prevalent with a subpopulation of intracellular Cx43 localized to lysosomes. In both cell types, Western blots revealed a notable increase in total cellular Cx43 in response to lysosome inhibitors. Interestingly, lactacystin inhibition of proteosomal degradation in MDA-MB-231 cells resulted in a marked increase in phosphorylated Cx43 at the expense of non-phosphorylated Cx43, and this change corresponded with an increase in "oversized" gap junction plaques. In BICR-M1Rk cells, lactacystin treatment partially prevented the BFA-induced loss of gap junctions. Together, our data suggests that lysosomes play a key role in not only degrading internalized gap junction in BICR-M1Rk cells but also in degrading Cx43 delivered from early secretory compartments to lysosomes in MDA-MB-231 cells. Overall proteasomal degradation regulates the stability of phosphorylated Cx43 and appears to promote the internalization of Cx43 from the cell surface. PMID- 12767975 TI - Inhibitory effects of basic or neutral phospholipid on acidic phospholipid mediated dissociation of adenine nucleotide bound to DnaA protein, the initiator of chromosomal DNA replication. AB - DnaA protein activity, the initiator of chromosomal DNA replication in bacteria, is regulated by acidic phospholipids such as phosphatidylglycerol (PG) or cardiolipin (CL) via facilitation of the exchange reaction of bound adenine nucleotide. Total lipid isolated from exponentially growing Staphylococcus aureus cells facilitated the release of ATP bound to S. aureus DnaA protein, whereas that from stationary phase cells was inert. Fractionation of total lipid from stationary phase cells revealed that the basic phospholipid, lysylphosphatidylglycerol (LPG), inhibited PG- or CL-facilitated release of ATP from DnaA protein. There was an increase in LPG concentration during the stationary phase. A fraction of the total lipid from stationary phase cells of an integrational deletion mprF mutant, in which LPG was lost, facilitated the release of ATP from DnaA protein. A zwitterionic phospholipid, phosphatidylethanolamine, also inhibited PG-facilitated ATP release. These results indicate that interaction of DnaA protein with acidic phospholipids might be regulated by changes in the phospholipid composition of the cell membrane at different growth stages. In addition, the mprF mutant exhibited an increased amount of origin per cell in vivo, suggesting that LPG is involved in regulating the cell cycle event(s). PMID- 12767976 TI - Oxidative stress contributes to arsenic-induced telomere attrition, chromosome instability, and apoptosis. AB - The environmental contaminant arsenic causes cancer, developmental retardation, and other degenerative diseases and, thus, is a serious health concern worldwide. Paradoxically, arsenic also may serve as an anti-tumor therapy, although the mechanisms of its antineoplastic effects remain unclear. Arsenic exerts its toxicity in part by generating reactive oxygen species. We show that arsenic induced oxidative stress promotes telomere attrition, chromosome end-to-end fusions, and apoptotic cell death. An antioxidant, N-acetylcysteine, effectively prevents arsenic-induced oxidative stress, telomere erosion, chromosome instability, and apoptosis, suggesting that increasing the intracellular antioxidant level may have preventive or therapeutic effects in arsenic-induced chromosome instability and genotoxicity. Embryos with shortened telomeres from late generation telomerase-deficient mice exhibit increased sensitivity to arsenic-induced oxidative damage, suggesting that telomere attrition mediates arsenic-induced apoptosis. Unexpectedly, arsenite did not cause chromosome end-to end fusions in telomerase RNA knockout mouse embryos despite progressively damaged telomeres and disrupting embryo viability. Together, these findings may explain why arsenic can initiate oxidative stress and telomere erosion, leading to apoptosis and anti-tumor therapy on the one hand and chromosome instability and carcinogenesis on the other. PMID- 12767977 TI - The closed structure of the MscS mechanosensitive channel. Cross-linking of single cysteine mutants. AB - Mechanosensitive channels must make a large conformational change during the transition from the closed to the open state. The crystal structure of the open form of the Escherichia coli MscS channel was recently solved and depicts a homoheptamer (1). In this study, cross-linking of site-specific cysteine substitutions demonstrates that residues up to 10-33 A apart in the crystal structure readily form disulfide bridges in the closed form and can also be cross linked by a 10-A linker. Cross-linking between adjacent subunits stabilizes the heptameric form of the channel providing biochemical evidence to support the crystal structure. The data are consistent with the published model (1) in that the membrane domain is highly flexible and that the closed to open transition may involve a significant displacement of transmembrane helices 1 and 2, possibly by as much as 30 A. The data are also consistent with significant flexibility of the cytoplasmic domain. PMID- 12767978 TI - Nucleosome sliding induced by the xMi-2 complex does not occur exclusively via a simple twist-diffusion mechanism. AB - ATP-dependent chromatin remodeling complexes can induce the translocation (sliding) of nucleosomes in cis along DNA, but the mechanism by which sliding occurs is not well defined. We previously presented evidence that sliding induced by the human SWI/SNF complex does not occur solely via a proposed "twist diffusion" mechanism whereby the DNA rotates about its helical axis without displacement from the surface of the nucleosome (Aoyagi, S., and Hayes, J. J. (2002) Mol. Cell. Biol. 22, 7484-7490). Here we examined whether the Xenopus Mi-2 nucleosome remodeling complex induces nucleosome sliding via a twist-diffusion mechanism with nucleosomes assembled onto DNA templates containing branched DNA structures expected to sterically hinder rotation of the DNA helix on the nucleosome surface. We find that the branched DNA-containing nucleosomes undergo xMi-2-catalyzed sliding at a rate and extent identical to that of nucleosomes assembled on native DNA fragments. These results indicate that both the hSWI/SNF and xMi-2 complexes induce nucleosome sliding via a mechanism(s) other than simple twist diffusion and are consistent with models in which the DNA largely maintains its rotational orientation with respect to the histone surface. PMID- 12767979 TI - Structural basis for distinctions between substrate and inhibitor specificities for feline immunodeficiency virus and human immunodeficiency virus proteases. AB - We used feline immunodeficiency virus (FIV) protease (PR) as a mutational framework to define determinants for the observed substrate and inhibitor specificity distinctions between FIV and human immunodeficiency virus (HIV) PRs. Multiple-substitution mutants were constructed by replacing the residues in and around the active site of FIV PR with the structurally equivalent residues of HIV 1 PR. Mutants included combinations of three critical regions (FIV numbering, with equivalent HIV numbering in superscript): I37(32)V in the active core region; N55(46)M, M56(47)I, and V59(50)I in the flap region; and L97(80)T, I98(81)P, Q99(82)V, P100(83)N, and L101(84)I in the 90s loop region. Significant alterations in specificity were observed, consistent with the involvement of these residues in determining the substrate-inhibitor specificity distinctions between FIV and HIV PRs. Two previously identified residues, I35 and I57 of FIV PR, were intolerant to substitution and yielded inactive PRs. Therefore, we attempted to recover the activity by introducing secondary mutations. The addition of G62(53)F and K63(54)I, located at the top of the flap and outside the active site, compensated for the activity lost in the I57(48)G substitution mutants. An additional two substitutions, D105(88)N and N88(74)T, facilitated recovery of activity in mutants that included the I35(30)D substitution. Determination of K(i) values of potent HIV-1 PR inhibitors against these mutants showed that inhibitor specificity paralleled that of HIV-1 PR. The findings indicate that maintenance of both substrate and inhibitor specificity is a function of interactions between residues both inside and outside the active site. Thus, mutations apparently peripheral to the active site can have a dramatic influence on inhibitor efficacy. PMID- 12767980 TI - Genome replication, virion secretion, and e antigen expression of naturally occurring hepatitis B virus core promoter mutants. AB - The core promoter mutants of hepatitis B virus (HBV) emerge as the dominant viral population at the late HBeAg and the anti-HBe stages of HBV infection, with the A1762T/G1764A substitutions as the hotspot mutations. The double core promoter mutations were found by many investigators to moderately enhance viral genome replication and reduce hepatitis B e antigen (HBeAg) expression. A much higher replication capacity was reported for a naturally occurring core promoter mutant implicated in the outbreak of fulminant hepatitis, which was caused by the neighboring C1766T/T1768A mutations instead. To systemically study the biological properties of naturally occurring core promoter mutants, we amplified full-length HBV genomes by PCR from sera of HBeAg(+) individuals infected with genotype A. All 12 HBV genomes derived from highly viremic sera (5 x 10(9) to 5.7 x 10(9) copies of viral genome/ml) harbored wild-type core promoter sequence, whereas 37 of 43 clones from low-viremia samples (0.2 x 10(7) to 4.6 x 10(7) copies/ml) were core promoter mutants. Of the 11 wild-type genomes and 14 core promoter mutants analyzed by transfection experiments in human hepatoma cell lines, 6 core promoter mutants but none of the wild-type genomes replicated at high levels. All had 1762/1764 mutations and an additional substitution at position 1753 (T to C), at position 1766 (C to T), or both. Moreover, these HBV clones varied greatly in their ability to secrete enveloped viral particles irrespective of the presence of core promoter mutations. High-replication clones with 1762/1764/1766 or 1753/1762/1764/1766 mutations expressed very low levels of HBeAg, whereas high replication clones with 1753/1762/1764 triple mutations expressed high levels of HBeAg. Experiments with site-directed mutants revealed that both 1762/1764/1766 and 1753/1762/1764/1766 mutations conferred significantly higher viral replication and lower HBeAg expression than 1762/1764 mutations alone, whereas the 1753/1762/1764 triple mutant displayed only mild reduction in HBeAg expression similar to the 1762/1764 mutant. Thus, core promoter mutations other than those at positions 1762 and 1764 can have major impact on viral DNA replication and HBeAg expression. PMID- 12767981 TI - Transplacental transfer and subsequent neonate utilization of herpes simplex virus-specific immunity are resilient to acute maternal stress. AB - Neonates are severely compromised in the ability to generate an immune response to pathogens and thus rely heavily on maternally derived immunity that is acquired by transplacental and transmammary means. The passive transfer of maternal herpes simplex virus (HSV)-specific antibody is critical in determining the outcome of neonatal HSV infection. In adults, psychological stress alters immune responsiveness via the increased level of corticosterone that is produced as a result of hypothalamic-pituitary-adrenal axis activation. Although the behavioral and neuroendocrine effects of pre- and postnatal stress-induced increases in corticosterone are well documented, the effects of maternal stress on the efficacy of prenatally transferred and neonatally developed viral immunity has yet to be addressed. By using a well-established prenatal restraint-and-light stress mouse model, we investigated the effects of increased maternal corticosterone on the passive transfer of total and HSV-specific immunoglobulin G (IgG) antibody and subsequent neonatal susceptibility to HSV infection. Serum corticosterone levels in pregnant mice were significantly increased in response to restraint-and-light stress, and fetuses derived from these stressed mice had significantly elevated levels of corticosterone. Despite the increases in corticosterone, the passive transfer of total and HSV-specific IgG antibody persisted and, in turn, protected the neonate from systemic viral spread. Therefore, prenatal stress did not increase the susceptibility of neonates to HSV type 2-associated mortality. These findings demonstrate the resiliency of the passive transfer of protective HSV-specific immunity under conditions of acute psychological stress. PMID- 12767982 TI - Complete sequence and genomic analysis of rhesus cytomegalovirus. AB - The complete DNA sequence of rhesus cytomegalovirus (RhCMV) strain 68-1 was determined with the whole-genome shotgun approach on virion DNA. The RhCMV genome is 221,459 bp in length and possesses a 49% G+C base composition. The genome contains 230 potential open reading frames (ORFs) of 100 or more codons that are arranged colinearly with counterparts of previously sequenced betaherpesviruses such as human cytomegalovirus (HCMV). Of the 230 RhCMV ORFs, 138 (60%) are homologous to known HCMV proteins. The conserved ORFs include the structural, replicative, and transcriptional regulatory proteins, immune evasion elements, G protein-coupled receptors, and immunoglobulin homologues. Interestingly, the RhCMV genome also contains sequences with homology to cyclooxygenase-2, an enzyme associated with inflammatory processes. Closer examination identified a series of candidate exons with the capacity to encode a full-length cyclooxygenase-2 protein. Counterparts of cyclooxygenase-2 have not been found in other sequenced herpesviruses. The availability of the complete RhCMV sequence along with the ability to grow RhCMV in vitro will facilitate the construction of recombinant viral strains for identifying viral determinants of CMV pathogenicity in the experimentally infected rhesus macaque and to the development of CMV as a vaccine vector. PMID- 12767983 TI - Human immunodeficiency virus type 1 entry inhibitors selected on living cells from a library of phage chemokines. AB - The chemokine receptors CCR5 and CXCR4 are promising non-virus-encoded targets for human immunodeficiency virus (HIV) therapy. We describe a selection procedure to isolate mutant forms of RANTES (CCL5) with antiviral activity considerably in excess of that of the native chemokine. The phage-displayed library of randomly mutated and N-terminally extended variants was screened by using live CCR5 expressing cells, and two of the selected mutants, P1 and P2, were further characterized. Both were significantly more potent HIV inhibitors than RANTES, with P2 being the most active (50% inhibitory concentration of 600 pM in a viral coat-mediated cell fusion assay, complete protection of target cells against primary HIV type 1 strains at a concentration of 10 nM). P2 resembles AOP-RANTES in that it is a superagonist of CCR5 and potently induces receptor sequestration. P1, while less potent than P2, has the advantage of significantly reduced signaling activity via CCR5 (30% of that of RANTES). Additionally, both P1 and P2 exhibit not only significantly increased affinity for CCR5 but also enhanced receptor selectivity, retaining only trace levels of signaling activity via CCR1 and CCR3. The phage chemokine approach that was successfully applied here could be adapted to other chemokine-chemokine receptor systems and used to further improve the first-generation mutants reported in this paper. PMID- 12767984 TI - Cytolysis by CCR5-using human immunodeficiency virus type 1 envelope glycoproteins is dependent on membrane fusion and can be inhibited by high levels of CD4 expression. AB - T-tropic (X4) and dualtropic (R5X4) human immunodeficiency virus type 1 (HIV-1) envelope glycoproteins kill primary and immortalized CD4(+) CXCR4(+) T cells by mechanisms involving membrane fusion. However, because much of HIV-1 infection in vivo is mediated by M-tropic (R5) viruses whose envelope glycoproteins use CCR5 as a coreceptor, we tested a panel of R5 and R5X4 envelope glycoproteins for their ability to lyse CCR5(+) target cells. As is the case for CXCR4(+) target cells, HIV-1 envelope glycoproteins expressed by single-round HIV-1 vectors killed transduced CD4(+) CCR5(+) cells in a membrane fusion-dependent manner. Furthermore, a CD4-independent R5 HIV-1 envelope glycoprotein was able to kill CD4-negative target cells expressing CCR5, demonstrating that CD4 is not intrinsically required for the induction of death. Interestingly, high levels of CD4 expression protected cells from lysis and syncytium formation mediated by the HIV-1 envelope glycoproteins. Immunoprecipitation experiments showed that high levels of CD4 coexpression inhibited proteolytic processing of the HIV-1 envelope glycoprotein precursor gp160. This inhibition could be overcome by decreasing the CD4 binding ability of gp120. Studies were also undertaken to investigate the ability of virion-bound HIV-1 envelope glycoproteins to kill primary CD4(+) T cells. However, neither X4 nor R5X4 envelope glycoproteins on noninfectious virions caused death in primary CD4(+) T cells. These results demonstrate that the interaction of CCR5 with R5 HIV-1 envelope glycoproteins capable of inducing membrane fusion leads to cell lysis; overexpression of CD4 can inhibit cell killing by limiting envelope glycoprotein processing. PMID- 12767985 TI - Varicella-zoster virus gene 66 transcription and translation in latently infected human Ganglia. AB - Latent infection with varicella-zoster virus (VZV) is characterized by restricted virus gene expression and the absence of virus production. Of the approximately 70 predicted VZV genes, only five (genes 4, 21, 29, 62, and 63) have been shown by multiple techniques to be transcribed during latency. IE62, the protein product of VZV gene 62, is the major immediate-early (IE) virus-encoded transactivator of viral gene transcription and plays a pivotal role in transactivating viral genes during lytic infection. The protein kinase (66-pk) encoded by VZV gene 66 phosphorylates IE62, resulting in cytoplasmic accumulation of IE62 that mitigates nuclear IE62-induced gene activation. Analysis of latently infected human trigeminal ganglia for 66-pk expression by reverse transcriptase dependent nested PCR, including DNA sequence analysis, in situ hybridization, and immunohistochemistry, revealed VZV open reading frame 66 to be a previously unrecognized latently expressed virus gene and suggests that prevention of IE62 import to the nucleus by VZV 66-pk phosphorylation is one possible mechanism by which VZV latency is maintained. PMID- 12767986 TI - Role of the human cytomegalovirus major immediate-early promoter's 19-base-pair repeat cyclic AMP-response element in acutely infected cells. AB - Prior studies have suggested a role of the five copies of the 19-bp-repeat cyclic AMP (cAMP)-response element (CRE) in major immediate-early (MIE) promoter activation, the rate-limiting step in human cytomegalovirus (HCMV) replication. We used two different HCMV genome modification strategies to test this hypothesis in acutely infected cells. We report the following: (i) the CREs do not govern basal levels of MIE promoter activity at a high or low multiplicity of infection (MOI) in human foreskin fibroblast (HFF)- or NTera2-derived neuronal cells; (ii) serum and virion components markedly increase MIE promoter-dependent transcription at a low multiplicity of infection (MOI), but this increase is not mediated by the CREs; (iii) forskolin stimulation of the cAMP signaling pathway induces a two- to threefold increase in MIE RNA levels in a CRE-specific manner at a low MOI in both HFF- and NTera2-derived neuronal cells; and (iv) the CREs do not regulate basal levels of HCMV DNA replication at a high or low MOI in HFF. Their presence does impart a forskolin-induced increase in viral DNA replication at a low MOI but only when basal levels of MIE promoter activity are experimentally diminished. In conclusion, the 19-bp-repeat CREs add to the robust MIE promoter activity that occurs in the acutely infected stimulated cells, although the CREs' greater role may be in other settings. PMID- 12767987 TI - Lectin-mediated retention of p62 facilitates p62-E1 heterodimerization in endoplasmic reticulum of Semliki Forest virus-infected cells. AB - The Semliki Forest virus (SFV) spike subunits p62 and E1 are made from a common coding unit in the order p62-E1. The proteins are separated by the host signal peptidase upon translocation into the endoplasmic reticulum (ER). Shortly thereafter, p62 and E1 form heterodimers. Heterodimerization preferentially occurs between subunits derived from the same translation product, so-called cis heterodimerization. As the p62 protein has the capacity to leave the ER in the absence of E1, it has been postulated that there exists a retention mechanism for the p62 protein, putatively at or near the translocon, in the ER in order to promote cis heterodimerization (B. U. Barth and H. Garoff, J. Virol. 71:7857 7865, 1997). Here we show that there exists such a mechanism, that it is at least in part mediated by the ER chaperones calnexin and calreticulin, and that the retention is important for efficient cis heterodimerization. PMID- 12767988 TI - Replicating parvoviruses that target colon cancer cells. AB - The wnt signaling pathway is constitutively activated in colon tumors by mutations in the adenomatous polyposis coli and beta-catenin genes. We have modified the minute virus of mice (MVM) P4 promoter to make it responsive to wnt signaling by inserting binding sites for the heterodimeric beta-catenin/Tcf transcription factor. In luciferase assays we can see up to 20-fold selectivity of Tcf mutant P4 promoters for cells with activated wnt signaling. Hybrid MVM/H-1 viruses containing Tcf mutant promoters were tested for NS1 expression, viral DNA replication, virus replication, and cytopathic effect on colon, lung, kidney, and cervical cancer cell lines. Activation of the wnt pathway by expression of Delta N-beta-catenin increased NS1 expression and viral burst size in 293T and H1299 lung cancer cells, showing that the Tcf mutant P4 promoter can respond to wnt signals in the context of the virus. Compared to the parental virus, the burst size of the Tcf mutant viruses was reduced at least 1,000-fold in H1299, 293T, NB324K, and HeLa cells, which have inactive wnt signaling pathways. The burst size and cytopathic effect of the Tcf viruses was near wild-type levels in SW480 and Isreco1 colon cancer cell lines, which have high Tcf activity. The high specificity of these viruses should permit the development of H-1 virus-based vectors which combine high safety and greater efficacy in cancer therapy. PMID- 12767989 TI - Protection against recurrent ocular herpes simplex virus type 1 disease after therapeutic vaccination of latently infected mice. AB - The potential of therapeutic vaccination of animals latently infected with herpes simplex virus type 1 (HSV-1) to enhance protective immunity to the virus and thereby reduce the incidence and severity of recurrent ocular disease was assessed in a mouse model. Mice latently infected with HSV-1 were vaccinated intranasally with a mixture of HSV-1 glycoproteins and recombinant Escherichia coli heat-labile enterotoxin B subunit (rEtxB) as an adjuvant. The systemic immune response induced was characterized by high levels of virus-specific immunoglobulin G1 (IgG1) in serum and very low levels of IgG2a. Mucosal immunity was demonstrated by high levels of IgA in eye and vaginal secretions. Proliferating T cells from lymph nodes of vaccinated animals produced higher levels of interleukin-10 (IL-10) than were produced by such cells from mock vaccinated animals. This profile suggests that vaccination of latently infected mice modulates the Th1-dominated proinflammatory response usually induced upon infection. After reactivation of latent virus by UV irradiation, vaccinated mice showed reduced viral shedding in tears as well as a reduction in the incidence of recurrent herpetic corneal epithelial disease and stromal disease compared with mock-vaccinated mice. Moreover, vaccinated mice developing recurrent ocular disease showed less severe signs and a quicker recovery rate. Spread of virus to other areas close to the eye, such as the eyelid, was also significantly reduced. Encephalitis occurred in a small percentage (11%) of mock-vaccinated mice, but vaccinated animals were completely protected from such disease. The possible immune mechanisms involved in protection against recurrent ocular herpetic disease in therapeutically vaccinated animals are discussed. PMID- 12767990 TI - Monocytes treated with human immunodeficiency virus Tat kill uninfected CD4(+) cells by a tumor necrosis factor-related apoptosis-induced ligand-mediated mechanism. AB - The human immunodeficiency virus (HIV) Tat protein has a critical role in viral transcription, but this study focuses on its additional role as an extracellular effector of lymphocyte cell death. It is well known that Tat induces tumor necrosis factor-related apoptosis-induced ligand (TRAIL) in peripheral blood mononuclear cells (PBMC), and we show that the majority of TRAIL is produced by the monocyte subset of PBMC. Human monocytes and U937 monoblastoid cells did not take up soluble HIV Tat-86, as T cells did, yet produced more TRAIL than did T cells. TRAIL secretion was induced by Tat and by a cysteine-rich peptide of Tat but not by sulfhydryl-modified Tat toxoid. Although there was only a slight increase in cell surface expression of TRAIL on monocytes, sufficient TRAIL was secreted to be toxic for T cells. The cytotoxicity of Tat-stimulated monocyte medium could be blocked by a TRAIL-neutralizing antibody. T cells treated with Tat did not secrete enough TRAIL to mediate cell death in our assay. Remarkably, uninfected T cells are more susceptible to TRAIL than are HIV-infected T cells. The production of TRAIL by Tat-stimulated monocytes provides a mechanism by which HIV infection can destroy uninfected bystander cells. PMID- 12767991 TI - Mechanisms of avian retroviral host range extension. AB - Alpharetroviruses provide a useful system for the study of the molecular mechanisms of host range and receptor interaction. These viruses can be divided into subgroups based on diverse receptor usage due to variability within the two host range determining regions, hr1 and hr2, in their envelope glycoprotein SU (gp85). In previous work, our laboratory described selection from a subgroup B avian sarcoma-leukosis virus of an extended-host-range variant (LT/SI) with two adjacent amino acid substitutions in hr1. This virus retains its ability to use the subgroup BD receptor but can also infect QT6/BD cells, which bear a related subgroup E receptor (R. A. Taplitz and J. M. Coffin, J. Virol 71:7814-7819, 1997). Here, we report further analysis of this unusual variant. First, one (L154S) of the two substitutions is sufficient for host range extension, while the other (T155I) does not alter host range. Second, these mutations extend host range to non-avian cell types, including human, dog, cat, mouse, rat, and hamster. Third, interference experiments imply that the mutants interact efficiently with the subgroup BD receptor and possibly the related subgroup E receptor, but they have another means of entry that is not dependent on these interactions. Fourth, binding studies indicate that the mutant SU proteins retain the ability to interact as monomers with subgroup BD and BDE receptors but only bind the subgroup E receptor in the context of an Env trimer. Further, the mutant SU proteins bind well to chicken cells but do not bind any better than wild-type subgroup B to QT6 or human cells, even though the corresponding viruses are capable of infecting these cells. PMID- 12767992 TI - Stem-loop III in the 5' untranslated region is a cis-acting element in bovine coronavirus defective interfering RNA replication. AB - Higher-order structures in the 5' untranslated region (UTR) of plus-strand RNA viruses are known in many cases to function as cis-acting elements in RNA translation, replication, or transcription. Here we describe evidence supporting the structure and a cis-acting function in defective interfering (DI) RNA replication of stem-loop III, the third of four predicted higher-order structures mapping within the 210-nucleotide (nt) 5' UTR of the 32-kb bovine coronavirus (BCoV) genome. Stem-loop III maps at nt 97 through 116, has a calculated free energy of -9.1 kcal/mol in the positive strand and -3.0 kcal/mol in the negative strand, and has associated with it beginning at nt 100 an open reading frame (ORF) potentially encoding an 8-amino-acid peptide. Stem-loop III is presumed to function in the positive strand, but its strand of action has not been established. Stem-loop III (i) shows phylogenetic conservation among group 2 coronaviruses and appears to have a homolog in coronavirus groups 1 and 3, (ii) has in all coronaviruses for which sequence is known a closely associated short, AUG-initiated intra-5' UTR ORF, (iii) is supported by enzyme structure-probing evidence in BCoV RNA, (iv) must maintain stem integrity for DI RNA replication in BCoV DI RNA, and (v) shows a positive correlation between maintenance of the short ORF and maximal DI RNA accumulation in BCoV DI RNA. These results indicate that stem-loop III in the BCoV 5' UTR is a cis-acting element for DI RNA replication and that its associated intra-5' UTR ORF may function to enhance replication. It is postulated that these two elements function similarly in the virus genome. PMID- 12767994 TI - Dual pressure from antiretroviral therapy and cell-mediated immune response on the human immunodeficiency virus type 1 protease gene. AB - Human immunodeficiency virus (HIV)-specific CD8(+) T-lymphocyte pressure can lead to the development of viral escape mutants, with consequent loss of immune control. Antiretroviral drugs also exert selection pressures on HIV, leading to the emergence of drug resistance mutations and increased levels of viral replication. We have determined a minimal epitope of HIV protease, amino acids 76 to 84, towards which a CD8(+) T-lymphocyte response is directed. This epitope, which is HLA-A2 restricted, includes two amino acids that commonly mutate (V82A and I84V) in the face of protease inhibitor therapy. Among 29 HIV-infected patients who were treated with protease inhibitors and who had developed resistance to these drugs, we show that the wild-type PR82V(76-84) epitope is commonly recognized by cytotoxic T lymphocytes (CTL) in HLA-A2-positive patients and that the CTL directed to this epitope are of high avidity. In contrast, the mutant PR82A(76-84) epitope is generally not recognized by wild-type-specific CTL, or when recognized it is of low to moderate avidity, suggesting that the protease inhibitor-selected V82A mutation acts both as a CTL and protease inhibitor escape mutant. Paradoxically, the absence of a mutation at position 82 was associated with the presence of a high-avidity CD8(+) T-cell response to the wild-type virus sequence. Our results indicate that both HIV type 1-specific CD8(+) T cells and antiretroviral drugs provide complex pressures on the same amino acid sequence of the HIV protease gene and, thus, can influence viral sequence evolution. PMID- 12767993 TI - Structure-function analysis of herpes simplex virus type 1 gD and gH-gL: clues from gDgH chimeras. AB - In alphaherpesviruses, glycoprotein B (gB), gD, gH, and gL are essential for virus entry. A replication-competent gL-null pseudorabies virus (PrV) (B. G. Klupp and T. C. Mettenleiter, J. Virol. 73:3014-3022, 1999) was shown to express a gDgH hybrid protein that could replace gD, gH, and gL in cell-cell fusion and null virus complementation assays. To study this phenomenon in herpes simplex virus type 1 (HSV-1), we constructed four gDgH chimeras, joining the first 308 gD amino acids to various gH N-terminal truncations. The chimeras were named for the first amino acid of gH at which each was truncated: 22, 259, 388, and 432. All chimeras were immunoprecipitated with both gD and gH antibodies to conformational epitopes. Normally, transport of gH to the cell surface requires gH-gL complex formation. Chimera 22 contains full-length gH fused to gD308. Unlike PrV gDgH, chimera 22 required gL for transport to the surface of transfected Vero cells. Interestingly, although chimera 259 failed to reach the cell surface, chimeras 388 and 432 exhibited gL-independent transport. To examine gD and gH domain function, each chimera was tested in cell-cell fusion and null virus complementation assays. Unlike PrV gDgH, none of the HSV-1 chimeras substituted for gL for fusion. Only chimera 22 was able to replace gH for fusion and could also replace either gH or gD in the complementation assay. Surprisingly, this chimera performed very poorly as a substitute for gD in the fusion assay despite its ability to complement gD-null virus and bind HSV entry receptors (HveA and nectin-1). Chimeras 388 and 432, which contain the same portion of gD as that in chimera 22, substituted for gD for fusion at 25 to 50% of wild-type levels. However, these chimeras functioned poorly in gD-null virus complementation assays. The results highlight the fact that these two functional assays are measuring two related but distinct processes. PMID- 12767995 TI - Specific inhibition of bovine viral diarrhea virus replicase. AB - Compound-1453 was identified and characterized as a specific inhibitor of bovine viral diarrhea virus (BVDV). The concentration of compound-1453 which results in 50% protection from virus-induced cytopathic effect is approximately 2.2 microM, with a therapeutic index of 60, and it is not active against a panel of RNA and DNA viruses. A time-of-addition experiment suggested that compound-1453 targets a stage of the viral life cycle after viral entry. To determine the target of compound-1453, resistant virus was generated. Resistant variants grew efficiently in the presence or absence of 33 micro M compound-1453 and exhibited replication efficiency in the presence of compound-1453 approximately 1,000-fold higher than that of the wild-type (wt) virus. Functional mapping and sequence analysis of resistant cDNAs revealed a single amino acid substitution (Glu to Gly) at residue 291 in the NS5B polymerase in all eight independently generated cDNA clones. Recombinant virus containing this single mutation retained the resistance phenotype and a replication efficiency similar to that of the original isolated resistant virus. Since compound-1453 did not inhibit BVDV polymerase activity in vitro (50% inhibitory concentration > 300 microM), we developed a membrane-based assay that consisted of a BVDV RNA replicase complex isolated from virus-infected cells. Compound-1453 inhibited the activity of the wt, but not the drug resistant, replicase in the membrane assay at concentrations similar to those observed in the viral infection assay. This work presents a novel inhibitor of a viral RNA-dependent RNA replicase. PMID- 12767996 TI - Kaposi's sarcoma-associated herpesvirus (human herpesvirus 8) contains hypoxia response elements: relevance to lytic induction by hypoxia. AB - Kaposi's sarcoma (KS)-associated herpesvirus (KSHV), also known as human herpesvirus 8, is an etiologic agent of KS, primary effusion lymphoma (PEL), and multicentric Castleman's disease. We recently demonstrated that hypoxia can induce lytic replication of KSHV in PEL cell lines. Hypoxia induces the accumulation of hypoxia-inducible factors (HIF), and we hypothesized that the KSHV genome may respond to hypoxia through functional hypoxia response elements (HREs). Here, we demonstrate the presence of at least two promoters within the KSHV genome that are activated by hypoxia or hypoxia mimics. One is in the promoter region of the gene for Rta, the main lytic switch gene, and the other is within the promoter region of ORF34, a lytic gene of unknown function. The ORF34 promoter contains three putative consensus HREs oriented in the direction of the gene. Dissection and site-directed mutagenesis studies confirmed that one of the HREs of the ORF34 promoter is functional. Under conditions of hypoxia, the ORF34 promoter was strongly upregulated by HIF-1 alpha and HIF-2 alpha. By contrast, the promoter of the gene for Rta appeared to be preferentially upregulated by HIF 2 alpha. Reverse transcription-PCR analysis revealed that specific messages for ORF34 and ORF50 are upregulated in BCBL-1 cells exposed to hypoxia. An HIF-1 binding and competition assay demonstrated that the HRE sequence from the ORF34 promoter can compete for HIF-1 alpha binding to an erythropoietin HRE oligonucleotide while a mutant sequence cannot. Thus, we demonstrated that a viral gene can be activated by hypoxia through activation of a functional viral HRE. To our knowledge, this is the first example of a functional HRE in a viral promoter. PMID- 12767997 TI - A transcriptionally active subgenomic promoter supports homologous crossovers in a plus-strand RNA virus. AB - Genetic RNA recombination plays an important role in viral evolution, but its molecular mechanism is not well understood. In this work we describe homologous RNA recombination activity that is supported by a subgenomic promoter (sgp) region in the RNA3 segment of brome mosaic bromovirus (BMV), a tripartite plus strand RNA virus. The crossover frequencies were determined by coinoculations with pairs of BMV RNA3 variants that carried a duplicated sgp region flanked by marker restriction sites. A region composed of the sgp core, a poly(A) tract, and an upstream enhancer supported homologous exchanges in 25% of the analyzed RNA3 progeny. However, mutations in the sgp core stopped both the transcription of the sgp RNA and homologous recombination. These data provide evidence for an association of RNA recombination with transcription. PMID- 12767998 TI - CAF-mediated human immunodeficiency virus (HIV) type 1 transcriptional inhibition is distinct from alpha-defensin-1 HIV inhibition. AB - CD8(+) T lymphocytes can inhibit human immunodeficiency virus type 1 (HIV-1) replication by secreting a soluble factor(s) known as CD8(+) T-lymphocyte antiviral factor (CAF). One site of CAF action is inhibition of HIV-1 RNA transcription, particularly at the step of long terminal repeat (LTR)-driven gene expression. The inhibitory effect of CAF on HIV-1 LTR activation is mediated through STAT1 activation. A recent study reports that alpha-defensins 1 to 3 account for CAF activity against HIV-1. Here, we address whether alpha-defensins, particularly alpha-defensin-1, contribute to CAF-mediated inhibition of HIV-1 transcription. Both recombinant alpha-defensin-1 and CAF derived from herpesvirus saimiri (HVS)-transformed CD8(+) cells inhibited HIV-1 infection and gene expression. For both factors, the inhibition of HIV-1 infection did not occur at the level of viral entry. Pretreatment of cells with alpha-defensin-1 followed by a washing out prior to infection blocked infection by HIV-1, indicating that direct inactivation of virions was not required for its inhibitory effect. In contrast to CAF, alpha-defensin-1 did not inhibit phorbol myristate acetate- or Tat-mediated HIV-1 LTR activation in a transient transfection system, nor did it activate STAT1 tyrosine phosphorylation. Furthermore, alpha-defensins 1 to 3 were below the level of detection in a panel of HVS-transformed CD8(+) cells with potent HIV-1 inhibitory activity and a neutralizing antibody against alpha defensins 1 to 3 did not reverse the inhibitory effect of CAF on HIV-1 gene expression in infected cells and on HIV-1 LTR activation in transfected cells. Taken together, our results suggest that alpha-defensin-1 inhibits HIV-1 infection following viral entry but that alpha-defensins 1 to 3 are not responsible for the HIV-1 transcriptional inhibition by CAF. PMID- 12767999 TI - Tomato yellow leaf curl Sardinia virus rep-derived resistance to homologous and heterologous geminiviruses occurs by different mechanisms and is overcome if virus-mediated transgene silencing is activated. AB - The replication-associated protein (Rep) of geminiviruses is involved in several biological processes brought about by the presence of distinct functional domains. Recently, we have exploited the multifunctional character of the Tomato yellow leaf curl Sardinia virus (TYLCSV) Rep to develop a molecular interference strategy to impair TYLCSV infection. We showed that transgenic expression of its N-terminal 210 amino acids (Rep-210) confers resistance to the homologous virus by inhibiting viral transcription and replication. We have now used biochemical and transgenic approaches to carry out a fuller investigation of the molecular resistance mechanisms in transgenic plants expressing Rep-210. We show that Rep 210 confers resistance through two distinct molecular mechanisms, depending on the challenging virus. Resistance to the homologous virus is achieved by the ability of Rep-210 to tightly inhibit C1 gene transcription, while that to heterologous virus is due to the interacting property of the Rep-210 oligomerization domain. Furthermore, we present evidence that in Rep-210 expressing plants, the duration of resistance is related to the ability of the challenging virus to shut off transgene expression by a posttranscriptional homology-dependent gene silencing mechanism. A model of Rep-210-mediated geminivirus resistance that takes transgene- and virus-mediated mechanisms into account is proposed. PMID- 12768000 TI - Cloning of an avian adeno-associated virus (AAAV) and generation of recombinant AAAV particles. AB - Recent studies have proposed that adeno-associated viruses (AAVs) are not evolutionarily linked to other mammalian autonomous parvoviruses but are more closely linked to the autonomous parvoviruses of birds. To better understand the relationship between primate and avian AAVs (AAAVs), we cloned and sequenced the genome of an AAAV (ATCC VR-865) and generated recombinant AAAV particles. The genome of AAAV is 4,694 nucleotides in length and has organization similar to that of other AAVs. The entire genome of AAAV displays 56 to 65% identity at the nucleotide level with the other known AAVs. The AAAV genome has inverted terminal repeats of 142 nucleotides, with the first 122 forming the characteristic T shaped palindromic structure. The putative Rep-binding element consists of a tandem (GAGY)(4) repeat, and the putative terminal resolution site (trs), CCGGT/CG, contains a single nucleotide substitution relative to the AAV(2) trs. The Rep open reading frame of AAAV displays 50 to 54% identity at the amino acid level with the other AAVs, with most of the diversity clustered at the carboxyl and amino termini. Comparison of the capsid proteins of AAAV and the primate dependoviruses indicate that divergent regions are localized to surface-exposed loops. Despite these sequence differences, we were able to produce recombinant AAAV particles carrying a lacZ reporter gene by cotransfection in 293T cells and were able to examine transduction efficiency in both chicken primary cells and several cell lines. Our findings indicate that AAAV is the most divergent AAV described to date but maintains all the characteristics unique to the genera of dependovirus. PMID- 12768002 TI - Determinants of increased replicative capacity of serially passaged simian immunodeficiency virus with nef deleted in rhesus monkeys. AB - Most rhesus macaques infected with simian immunodeficiency virus SIVmac239 with nef deleted (either Delta nef or Delta nef Delta vpr Delta US [Delta 3]) control viral replication and do not progress to AIDS. Some monkeys, however, develop moderate viral load set points and progress to AIDS. When simian immunodeficiency viruses (SIVs) recovered from two such animals (one Delta nef and the other Delta 3) were serially passaged in rhesus monkeys, the SIVs derived from both lineages were found to consistently induce moderate viral loads and disease progression. Analysis of viral sequences in the serially passaged derivatives revealed interesting changes in three regions: (i) an unusually high number of predicted amino acid changes (12 to 14) in the cytoplasmic domain of gp41, most of which were in regions that are usually conserved; these changes were observed in both lineages; (ii) an extreme shortening of nef sequences in the region of overlap with U3; these changes were observed in both lineages; and (iii) duplication of the NF-kappa B binding site in one lineage only. Neither the polymorphic gp41 changes alone nor the U3 deletion alone appeared to be responsible for increased replicative capacity because recombinant SIVmac239 Delta nef, engineered to contain either of these changes, induced moderate viral loads in only one of six monkeys. However, five of six monkeys infected with recombinant SIVmac239 Delta nef containing both TM and U3 changes did develop persisting moderate viral loads. These genetic changes did not increase lymphoid cell-activating properties in the monkey interleukin-2-dependent T-cell line 221, but the gp41 changes did increase the fusogenic activity of the SIV envelope two- to threefold. These results delineate sequence changes in SIV that can compensate for the loss of the nef gene to partially restore replicative and pathogenic potential in rhesus monkeys. PMID- 12768001 TI - Turnover of env variable region 1 and 2 genotypes in subjects with late-stage human immunodeficiency virus type 1 infection. AB - The env gene of human immunodeficiency virus type 1 (HIV-1) includes some of the most genetically diverse regions of the viral genome, which are called variable regions 1 through 5 (V1 through V5). We have developed a heteroduplex tracking assay to detect changes in variable regions 1 and 2 of env (V1/V2-HTA). Using sequences from two molecular clones as probes, we have studied the nature of longitudinal virus population changes in a cohort of HIV-1-infected subjects. Viral sequences present in 21 subjects with late-stage HIV-1 infection were initially screened for stability of the virus population by V1/V2-HTA. The virus populations at entry comprised an average of five coexisting V1/V2 genotypic variants (as identified by HTA). Eight of the 21 subjects were examined in detail because of the dynamic behavior of their env variants over an approximately 9 month period. In each of these cases we detected a single discrete transition of V1/V2 genotypes based on monthly sampling. The major V1/V2 genotypes (those present at >10% abundance) from the eight subjects were cloned and sequenced to define the nature of V1/V2 variability associated with a discrete transition. Based on a comparison of V1/V2 genotypic variants present at entry with the newly emerged variants we categorized the newly emerged variants into two groups: variants without length differences and variants with length differences. Variants without length differences had fewer nucleotide substitutions, with the changes biased to either V1 or V2, suggestive of recent evolutionary events. Variants with length differences included ones with larger numbers of changes that were distributed, suggestive of recall of older genotypes. Most length differences were located in domains where the codon motif AVT (V = A, G, C) had become enriched and fixed. Finally, recombination events were detected in two subjects, one of which resulted in the reassortment of V1 and V2 regions. We suggest that turnover in V1/V2 populations was largely driven by selection on either V1 or V2 and that escape was accomplished either through changes focused in the region under selection or by the appearance of a highly divergent variant. PMID- 12768003 TI - Herpes simplex virus glycoprotein K, but not its syncytial allele, inhibits cell cell fusion mediated by the four fusogenic glycoproteins, gD, gB, gH, and gL. AB - A Myc epitope was inserted at residue 283 of herpes simplex virus type 1 (HSV-1) glycoprotein K (gK), a position previously shown not to interfere with gK activity. The Myc-tagged gK localized predominantly to the endoplasmic reticulum, both in uninfected and in HSV-infected cells. gK, coexpressed with the four HSV fusogenic glycoproteins, gD, gB, gH, and gL, inhibited cell-cell fusion. The effect was partially dose dependent and was observed both in baby hamster kidney (BHK) and in Vero cells, indicating that the antifusion activity of gK may be cell line independent. The antifusion activity of gK did not require viral proteins other than the four fusogenic glycoproteins. A syncytial (syn) allele of gK (syn-gK) carrying the A40V substitution present in HSV-1(MP) did not block fusion to the extent seen with the wild-type (wt) gK, indicating that the syn mutation ablated, at least in part, the antifusogenic activity of wt gK. We conclude that gK is part of the mechanism whereby HSV negatively regulates its own fusion activity. Its effect accounts for the notion that cells infected with wt HSV do not fuse with adjacent, uninfected cells into multinucleated giant cells or syncytia. gK may also function to preclude fusion between virion envelope and the virion-encasing vesicles during virus transport to the extracellular compartment, thus preventing nucleocapsid de-envelopment in the cytoplasm. PMID- 12768004 TI - Follicular dendritic cell dedifferentiation by treatment with an inhibitor of the lymphotoxin pathway dramatically reduces scrapie susceptibility. AB - Transmissible spongiform encephalopathies (TSEs) may be acquired peripherally, in which case infectivity usually accumulates in lymphoid tissues before dissemination to the nervous system. Studies of mouse scrapie models have shown that mature follicular dendritic cells (FDCs), expressing the host prion protein (PrP(c)), are critical for replication of infection in lymphoid tissues and subsequent neuroinvasion. Since FDCs require lymphotoxin signals from B lymphocytes to maintain their differentiated state, blockade of this stimulation with a lymphotoxin beta receptor-immunoglobulin fusion protein (LT beta R-Ig) leads to their temporary dedifferentiation. Here, a single treatment with LT beta R-Ig before intraperitoneal scrapie inoculation blocked the early accumulation of infectivity and disease-specific PrP (PrP(Sc)) within the spleen and substantially reduced disease susceptibility. These effects coincided with an absence of FDCs in the spleen for ca. 28 days after treatment. Although the period of FDC dedifferentiation was extended to at least 49 days by consecutive LT beta R-Ig treatments, this had little added protective benefit after injection with a moderate dose of scrapie. We also demonstrate that mature FDCs are critical for the transmission of scrapie from the gastrointestinal tract. Treatment with LT beta R-Ig before oral scrapie inoculation blocked PrP(Sc) accumulation in Peyer's patches and mesenteric lymph nodes and prevented neuroinvasion. However, treatment 14 days after oral inoculation did not affect survival time or susceptibility, suggesting that infectivity may have already spread to the peripheral nervous system. Although manipulation of FDCs may offer a potential approach for early intervention in peripherally acquired TSEs, these data suggest that the duration of the treatment window may vary widely depending on the route of exposure. PMID- 12768005 TI - Expression of simian immunodeficiency virus (SIV) nef in astrocytes during acute and terminal infection and requirement of nef for optimal replication of neurovirulent SIV in vitro. AB - As the most numerous cells in the brain, astrocytes play a critical role in maintaining central nervous system homeostasis, and therefore, infection of astrocytes by human immunodeficiency virus (HIV) or simian immunodeficiency virus (SIV) in vivo could have important consequences for the development of HIV encephalitis. In this study, we establish that astrocytes are infected in macaques during acute SIV infection (10 days postinoculation) and during terminal infection when there is evidence of SIV-induced encephalitis. Additionally, with primary adult rhesus macaque astrocytes in vitro, we demonstrate that the macrophage-tropic, neurovirulent viruses SIV/17E-Br and SIV/17E-Fr replicate efficiently in astrocytes, while the lymphocyte-tropic, nonneurovirulent virus SIV(mac)239 open-nef does not establish productive infection. Furthermore, aminoxypentane-RANTES abolishes virus replication, suggesting that these SIV strains utilize the chemokine receptor CCR5 for entry into astrocytes. Importantly, we show that SIV Nef is required for optimal replication in primary rhesus macaque astrocytes and that normalizing input virus by particle number rather than by infectivity reveals a disparity between the ability of a Nef deficient virus and a virus encoding a nonmyristoylated form of Nef to replicate in these central nervous system cells. Since the myristoylated form of Nef has been implicated in functions such as CD4 and major histocompatibility complex I downregulation, kinase association, and enhancement of virion infectivity, these data suggest that an as yet unidentified function of Nef may exist to facilitate SIV replication in astrocytes that may have important implications for in vivo pathogenesis. PMID- 12768006 TI - Comprehensive analysis of human immunodeficiency virus type 1 (HIV-1)-specific gamma interferon-secreting CD8+ T cells in primary HIV-1 infection. AB - Human immunodeficiency virus type 1 (HIV-1)-specific CD8(+) T cells provide an important defense in controlling HIV-1 replication, particularly following acquisition of infection. To delineate the breadth and potency of these responses in patients upon initial presentation and before treatment, we determined the fine specificities and frequencies of gamma interferon (IFN-gamma)-secreting CD8(+) T cells recognizing all HIV-1 proteins in patients with primary infection. In these subjects, the earliest detected responses were directed predominantly against Nef, Tat, Vpr, and Env. Tat- and Vpr-specific CD8(+) T cells accounted for the greatest frequencies of mean IFN-gamma spot-forming cells (SFC). Nef specific responses (10 of 21) were more commonly detected. A mean of 2.3 epitopes were recognized with various avidities per subject, and the number increased with the duration of infection (R = 0.47, P = 0.031). The mean frequency of CD8(+) T cells (985 SFC/10(6) peripheral blood mononuclear cells) correlated with the number of epitopes recognized (R = 0.84, P < 0.0001) and the number of HLA restricting alleles (R = 0.79, P < 0.0001). Neither the total SFC frequencies nor the number of epitopes recognized correlated with the concurrent plasma viral load. Seventeen novel epitopes were identified, four of which were restricted to HLA alleles (A23 and B72) that are common among African descendents. Thus, primary HIV-1 infection induces strong CD8(+)-T-cell immunity whose specificities broaden over time, but their frequencies and breadth do not correlate with HIV-1 containment when examined concurrently. Many novel epitopes, particularly directed to Nef, Tat, and Env, and frequently with unique HLA restrictions, merit further consideration in vaccine design. PMID- 12768007 TI - Characterization of a novel simian immunodeficiency virus (SIVmonNG1) genome sequence from a mona monkey (Cercopithecus mona). AB - A novel simian immunodeficiency virus (SIV) sequence has been recovered from RNA extracted from the serum of a mona monkey (Cercopithecus mona) wild born in Nigeria. The sequence was obtained by using novel generic (degenerate) PCR primers and spans from two-thirds into the gag gene to the 3' poly(A) tail of the SIVmonNG1 RNA genome. Analysis of the open reading frames revealed that the SIVmonNG1 genome codes for a Vpu protein, in addition to Gag, Pol, Vif, Vpr, Tat, Rev, Env, and Nef proteins. Previously, only lentiviruses infecting humans (human immunodeficiency virus type 1 [HIV-1]) and chimpanzees (SIVcpz) were known to have a vpu gene; more recently, this has also been found in SIVgsn from Cercopithecus nictitans. Overall, SIVmonNG1 most closely resembles SIVgsn: the env gene sequence groups with HIV-1/SIVcpz env sequences, whereas the pol gene sequence clusters closely with the pol sequence of SIVsyk from Cercopithecus albogaris. By bootscanning and similarity plotting, the first half of pol resembles SIVsyk, whereas the latter part is closer to SIVcol from Colobus guereza. The similarities between the complex mosaic genomes of SIVmonNG1 and SIVgsn are consistent with a shared or common lineage. These data further highlight the intricate nature of the relationships between the SIVs from different primate species and will be helpful for unraveling these associations. PMID- 12768008 TI - The differential ability of HLA B*5701+ long-term nonprogressors and progressors to restrict human immunodeficiency virus replication is not caused by loss of recognition of autologous viral gag sequences. AB - Although the HLA B(*)5701 class I allele is highly overrepresented among human immunodeficiency virus (HIV)-infected long-term nonprogressors (LTNPs), it is also present at the expected frequency (11%) in patients with progressive HIV infection. Whether B57(+) progressors lack restriction of viral replication because of escape from recognition of highly immunodominant B57-restricted gag epitopes by CD8(+) T cells remains unknown. In this report, we investigate the association between restriction of virus replication and recognition of autologous virus sequences in 27 B(*)57(+) patients (10 LTNPs and 17 progressors). Amplification and direct sequencing of single molecules of viral cDNA or proviral DNA revealed low frequencies of genetic variations in these regions of gag. Furthermore, CD8(+) T-cell recognition of autologous viral variants was preserved in most cases. In two patients, responses to autologous viral variants were not demonstrable at one epitope. By using a novel technique to isolate primary CD4(+) T cells expressing autologous viral gene products, it was found that 1 to 13% of CD8(+) T cells were able to respond to these cells by gamma interferon production. In conclusion, escape-conferring mutations occur infrequently within immunodominant B57-restricted gag epitopes and are not the primary mechanism of virus evasion from immune control in B(*)5701(+) HIV infected patients. Qualitative features of the virus-specific CD8(+) T-cell response not measured by current assays remain the most likely determinants of the differential abilities of HLA B(*)5701(+) LTNPs and progressors to restrict virus replication. PMID- 12768009 TI - Human immunodeficiency virus type 1 envelope-mediated neuronal death: uncoupling of viral replication and neurotoxicity. AB - Although brain tissue from patients with human immunodeficiency virus (HIV) and/or AIDS is consistently infected by HIV type 1 (HIV-1), only 20 to 30% of patients exhibit clinical or neuropathological evidence of brain injury. Extensive HIV-1 sequence diversity is present in the brain, which may account in part for the variability in the occurrence of HIV-induced brain disease. Neurological injury caused by HIV-1 is mediated directly by neurotoxic viral proteins or indirectly through excess production of host molecules by infected or activated glial cells. To elucidate the relationship between HIV-1 infection and neuronal death, we examined the neurotoxic effects of supernatants from human 293T cells or macrophages expressing recombinant HIV-1 virions or gp120 proteins containing the V1V3 or C2V3 envelope region from non-clade B, brain-derived HIV-1 sequences. Neurotoxicity was measured separately as apoptosis or total neuronal death, with apoptosis representing 30 to 80% of the total neuron death observed, depending on the individual virus. In addition, neurotoxicity was dependent on expression of HIV-1 gp120 and could be blocked by anti-gp120 antibodies, as well as by antibodies to the human CCR5 and CXCR4 chemokine receptors. Despite extensive sequence diversity in the recombinant envelope region (V1V3 or C2V3), there was limited variation in the neurotoxicity induced by supernatants from transfected 293T cells. Conversely, supernatants from infected macrophages caused a broader range of neurotoxicity levels that depended on each virus and was independent of the replicative ability of the virus. These findings underscore the importance of HIV-1 envelope protein expression in neurotoxic pathways associated with HIV-induced brain disease and highlight the envelope as a target for neuroprotective therapeutic interventions. PMID- 12768010 TI - Fusion deficiency induced by mutations at the dimer interface in the Newcastle disease virus hemagglutinin-neuraminidase is due to a temperature-dependent defect in receptor binding. AB - The tetrameric paramyxovirus hemagglutinin-neuraminidase (HN) protein mediates attachment to sialic acid-containing receptors as well as cleavage of the same moiety via its neuraminidase (NA) activity. The X-ray crystallographic structure of an HN dimer from Newcastle disease virus (NDV) suggests that a single site in two different conformations mediates both of these activities. This conformational change is predicted to involve an alteration in the association between monomers in each HN dimer and to be part of a series of changes in the structure of HN that link its recognition of receptors to the activation of the other viral surface glycoprotein, the fusion protein. To explore the importance of the dimer interface to HN function, we performed a site-directed mutational analysis of residues in a domain defined by residues 218 to 226 at the most membrane-proximal part of the dimer interface in the globular head. Proteins carrying substitutions for residues F220, S222, and L224 in this domain were fusion deficient. However, this fusion deficiency was not due to a direct effect of the mutations on fusion. Rather, the fusion defect was due to a severely impaired ability to mediate receptor recognition at 37 degrees C, a phenotype that is not attributable to a change in NA activity. Since each of these mutated proteins efficiently mediated attachment in the cold, it was also not due to an inherent inability of the mutated proteins to recognize receptors. Instead, the interface mutations acted by weakening the interaction between HN and its receptor(s). The phenotype of these mutants correlates with the disruption of intermonomer subunit interactions. PMID- 12768011 TI - Sequence and structure of human rhinoviruses reveal the basis of receptor discrimination. AB - The sequences of the capsid protein VP1 of all minor receptor group human rhinoviruses were determined. A phylogenetic analysis revealed that minor group HRVs were not more related to each other than to the nine major group HRVs whose sequences are known. Examination of the surface exposed amino acid residues of HRV1A and HRV2, whose X-ray structures are available, and that of three dimensional models computed for the remaining eight minor group HRVs indicated a pattern of positively charged residues within the region, which, in HRV2, was shown to be the binding site of the very-low-density lipoprotein (VLDL) receptor. A lysine in the HI loop of VP1 (K224 in HRV2) is strictly conserved within the minor group. It lies in the middle of the footprint of a single repeat of the VLDL receptor on HRV2. Major group virus serotypes exhibit mostly negative charges at the corresponding positions and do not bind the negatively charged VLDL receptor, presumably because of charge repulsion. PMID- 12768012 TI - Targeting of the human immunodeficiency virus type 1 envelope to the trans-Golgi network through binding to TIP47 is required for env incorporation into virions and infectivity. AB - Here, we report that human immunodeficiency virus type 1 (HIV-1) Env glycoprotein is located mainly in the trans-Golgi network (TGN) due to determinants present in the cytoplasmic domain of the transmembrane gp41 glycoprotein (TMgp41). Internalization assays demonstrated that Env present at the cell surface returns to the TGN. We found that the cytoplasmic domain of TMgp41 binds to TIP47, a protein required for the transport of mannose-6-phosphate receptors from endosomes to the TGN. Overexpression of a mutant of TIP47 affected the transport of Env from endosomes to the TGN. Retrograde transport of Env to the TGN requires a Y(802)W(803) diaromatic motif present in the TMgp41 cytoplasmic domain. Mutation of this motif abolished both targeting to the TGN as well as interaction with TIP47. These data support the view that binding of TIP47 to HIV-1 Env facilitates its delivery to the TGN. Lastly, we show that virus mutated in the Y(802)W(803) motif is poorly infectious and presents a defect in Env incorporation, supporting a model in which retrograde transport of Env is implicated in the optimization of fully infectious HIV-1 production. PMID- 12768014 TI - The PDZ ligand domain of the human papillomavirus type 16 E6 protein is required for E6's induction of epithelial hyperplasia in vivo. AB - Human papillomaviruses (HPVs) are the causative agent of warts. Infections with high-risk HPVs are associated with anogenital and head and neck cancers. One of the viral genes responsible for HPV's oncogenic activity is E6. Mice expressing the HPV-16 E6 protein in their epidermis (K14E6(WT)) develop epithelial hyperplasia and squamous carcinomas. Numerous cellular proteins interact with E6, some of which can be grouped based on common amino acid motifs in their E6 binding domains. One such group, the PDZ partners, including hDLG, hSCRIBBLE, MUPP1, and MAGI, bind to the carboxy-terminal four amino acids of E6 through their PDZ domains. E6's interaction with the PDZ partners leads to their degradation. Additionally, E6's binding to PDZ proteins has been correlated with its ability to transform baby rat kidney cells in tissue culture and to confer tumorigenicity onto cells in xenograft experiments. To address whether the ability of E6 to bind PDZ domain partners is necessary for E6 to confer epithelial hyperproliferation in vivo, we generated transgenic mice that express in stratified squamous epithelia a mutant of E6 lacking the last six amino acids at its carboxyl terminus, E6(Delta 146-151), from the human keratin 14 (K14) promoter. The K14E6(Delta 146-151) mice exhibit a radiation response similar to that of the K14E6(WT) mice, demonstrating that this protein, as predicted, retains an ability to inactivate p53. However, the K14E6(Delta 146-151) mice fail to display epithelial hyperplasia. These results indicate that an interaction of E6 with PDZ partners is necessary for its induction of epithelial hyperplasia. PMID- 12768013 TI - EBNA1 partitions Epstein-Barr virus plasmids in yeast cells by attaching to human EBNA1-binding protein 2 on mitotic chromosomes. AB - Epstein-Barr virus (EBV) episomal genomes are stably maintained in human cells and are partitioned during cell division by mitotic chromosome attachment. Partitioning is mediated by the viral EBNA1 protein, which binds both the EBV segregation element (FR) and a mitotic chromosomal component. We previously showed that the segregation of EBV-based plasmids can be reconstituted in Saccharomyces cerevisiae and is absolutely dependent on EBNA1, the EBV FR sequence, and the human EBNA1-binding protein 2 (EBP2). We have now used this yeast system to elucidate the functional contribution of human EBP2 to EBNA1 mediated plasmid partitioning. Human EBP2 was found to attach to yeast mitotic chromosomes in a cell cycle-dependent manner and cause EBNA1 to associate with the mitotic chromosomes. The domain of human EBP2 that binds both yeast and human chromosomes was mapped and shown to be functionally distinct from the EBNA1 binding domain. The functionality and localization of human EBP2 mutants and fusion proteins indicated that the attachment of EBNA1 to mitotic chromosomes is crucial for EBV plasmid segregation in S. cerevisiae, as it is in humans, and that this is the contribution of human EBP2. The results also indicate that plasmid segregation in S. cerevisiae can occur through chromosome attachment. PMID- 12768016 TI - Varicella-zoster virus DNA in cells isolated from human trigeminal ganglia. AB - To determine the type of cell(s) that contain latent varicella-zoster virus (VZV) DNA, we prepared pure populations of neurons and satellite cells from trigeminal ganglia of 18 humans who had previously had a VZV infection. VZV DNA was present in 34 of 2,226 neurons (1.5%) and in none of 20,700 satellite cells. There was an average of 4.7 (range of 2 to 9) copies of VZV DNA per latently infected neuron. Latent VZV DNA was primarily present in large neurons, whereas the size distribution of herpes simplex virus DNA was markedly different. PMID- 12768015 TI - A novel human antibody against human immunodeficiency virus type 1 gp120 is V1, V2, and V3 loop dependent and helps delimit the epitope of the broadly neutralizing antibody immunoglobulin G1 b12. AB - The V1/V2 and V3 loops are proximal to the CD4 binding site (CD4bs) of human immunodeficiency virus type 1 (HIV-1) gp120 and undergo conformational change upon CD4 receptor engagement by the HIV-1 envelope spike. Nearly all of the reported monoclonal antibodies (MAbs) against the CD4bs exhibit a very limited capacity to neutralize HIV-1. However, one such human MAb, immunoglobulin G1 (IgG1) b12, is uniquely able to neutralize primary isolates across subtypes with considerable potency. The molecular basis for the anti-HIV-1 activity of b12 is not fully understood but is relevant to vaccine design. Here we describe a novel human MAb, 4KG5, whose binding to monomeric gp120 is moderately enhanced by IgG1 b12. In sharp contrast, 4KG5 binding to gp120 is inhibited by soluble CD4 (sCD4) and by all other (n = 14) anti-CD4bs MAbs tested. 4KG5 is unable to recognize gp120 in which either V1, V2, or V3 has been deleted, and MAbs against the V2 or V3 loops inhibit the binding of 4KG5 to gp120. Moreover, 4KG5 is able to inhibit the binding of the CD4-induced MAbs 17b and X5 in the absence of sCD4, whereas 17b and X5 only weakly inhibit the binding of 4KG5 to gp120. Mutagenesis of gp120 provides further evidence of a discontinuous epitope of 4KG5 that is formed by the V1/V2 loop, the V3 loop, and a portion of the bridging sheet (C4). 4KG5 was isolated as a single-chain Fv from a phage display library constructed from the bone marrow of an HIV-1-seropositive subject (FDA2) whose serum neutralizes HIV-1 across subtypes. Despite its source, we observed no significant neutralization with 4KG5 against the autologous (R2) virus and several other strains of HIV-1. The results suggest a model in which antibody access to the CD4bs on the envelope spike of HIV-1 is restricted by the orientation and/or dynamics of the V1/V2 and V3 loops, and b12 avoids these restrictions. PMID- 12768017 TI - Characterization of H9 subtype influenza viruses from the ducks of southern China: a candidate for the next influenza pandemic in humans? AB - A current view of the emergence of pandemic influenza viruses envisages a gene flow from the aquatic avian reservoir to humans via reassortment in pigs, the hypothetical "mixing vessel." Understanding arising from recent H5N1 influenza outbreaks in Hong Kong since 1997 and the isolation of avian H9N2 virus from humans raises alternative options for the emergence of a new pandemic virus. Here we report that H9N2 influenza viruses established in terrestrial poultry in southern China are transmitted back to domestic ducks, in which the viruses generate multiple reassortants. These novel H9N2 viruses are double or even triple reassortants that have amino acid signatures in their hemagglutinin, indicating their potential to directly infect humans. Some of them contain gene segments that are closely related to those of A/Hong Kong/156/97 (H5N1/97, H5N1) or A/Quail/Hong Kong/G1/97 (G1-like, H9N2). More importantly, some of their internal genes are closely related to those of novel H5N1 viruses isolated during the outbreak in Hong Kong in 2001. This study reveals a two-way transmission of influenza virus between terrestrial and aquatic birds that facilitates the generation of novel reassortant H9N2 influenza viruses. Such reassortants may directly or indirectly play a role in the emergence of the next pandemic virus. PMID- 12768019 TI - Human cytomegalovirus infection inhibits tumor necrosis factor alpha (TNF-alpha) signaling by targeting the 55-kilodalton TNF-alpha receptor. AB - Infection with human cytomegalovirus (HCMV) results in complex interactions between viral and cellular factors which perturb many cellular functions. HCMV is known to target the cell cycle, cellular transcription, and immunoregulation, and it is believed that this optimizes the cellular environment for viral DNA replication during productive infection or during carriage in the latently infected host. Here, we show that HCMV infection also prevents external signaling to the cell by disrupting the function of TNFRI, the 55-kDa receptor for tumor necrosis factor alpha (TNF-alpha), one of the receptors for a potent cytokine involved in eliciting a wide spectrum of cellular responses, including antiviral responses. HCMV infection of fully permissive differentiated monocytic cell lines and U373 cells resulted in a reduction in cell surface expression of TNFRI. The reduction appeared to be due to relocalization of TNFRI from the cell surface and was reflected in the elimination of TNF-alpha-induced Jun kinase activity. Analysis of specific phases of infection suggested that viral early gene products were responsible for this relocalization. However, a mutant HCMV in which all viral gene products known to be involved in down-regulation of major histocompatibility complex (MHC) class I were deleted still resulted in relocalization of TNFRI. Consequently, TNFRI relocalization by HCMV appears to be mediated by a novel viral early function not involved in down-regulation of cell surface MHC class I expression. We suggest that upon infection, HCMV isolates the cell from host-mediated signals, forcing the cell to respond only to virus specific signals which optimize the cell for virus production and effect proviral responses from bystander cells. PMID- 12768018 TI - Identification of amino acid residues in the capsid proteins of adeno-associated virus type 2 that contribute to heparan sulfate proteoglycan binding. AB - The adeno-associated virus type 2 (AAV2) uses heparan sulfate proteoglycan (HSPG) as its primary cellular receptor. In order to identify amino acids within the capsid of AAV2 that contribute to HSPG association, we used biochemical information about heparin and heparin sulfate, AAV serotype protein sequence alignments, and data from previous capsid studies to select residues for mutagenesis. Charged-to-alanine substitution mutagenesis was performed on individual residues and combinations of basic residues for the production and purification of recombinant viruses that contained a green fluorescent protein (GFP) reporter gene cassette. Intact capsids were assayed for their ability to bind to heparin-agarose in vitro, and virions that packaged DNA were assayed for their ability to transduce normally permissive cell lines. We found that mutation of arginine residues at position 585 or 588 eliminated binding to heparin agarose. Mutation of residues R484, R487, and K532 showed partial binding to heparin-agarose. We observed a general correlation between heparin-agarose binding and infectivity as measured by GFP transduction; however, a subset of mutants that partially bound heparin-agarose (R484A and K532A) were completely noninfectious, suggesting that they had additional blocks to infectivity that were unrelated to heparin binding. Conservative mutation of positions R585 and R588 to lysine slightly reduced heparin-agarose binding and had comparable effects on infectivity. Substitution of AAV2 residues 585 through 590 into a location predicted to be structurally equivalent in AAV5 generated a hybrid virus that bound to heparin-agarose efficiently and was able to package DNA but was noninfectious. Taken together, our results suggest that residues R585 and R588 are primarily responsible for heparin sulfate binding and that mutation of these residues has little effect on other aspects of the viral life cycle. Interactive computer graphics examination of the AAV2 VP3 atomic coordinates revealed that residues which contribute to heparin binding formed a cluster of five basic amino acids that presented toward the icosahedral threefold axis from the surrounding spike protrusion. Three other kinds of mutants were identified. Mutants R459A, H509A, and H526A/K527A bound heparin at levels comparable to that of wild-type virus but were defective for transduction. Another mutant, H358A, was defective for capsid assembly. Finally, an R459A mutant produced significantly lower levels of full capsids, suggesting a packaging defect. PMID- 12768020 TI - Generation of transduction-competent retroviral vectors by infection with a single hybrid vaccinia virus. AB - Recombinant vaccinia viruses that express defective retroviral vectors upon a single infection event in normal host cells were constructed. The gag-pol and envelope genes and a retroviral vector unit were inserted as vaccinia virus promoter-controlled transcription units at three separate loci. The triple recombinant virus was used to infect such diverse cell types as monkey and rabbit kidney, human lung, and primary chicken cells, resulting in the production of transduction-competent defective retroviral vectors. Infection of Chinese hamster ovary cells, which are nonpermissive for vaccinia virus replication, also resulted in production of retroviral vectors and secondary permanent transduction of the host cells. Since vaccinia virus supports the expression of cytotoxic proteins, the vesicular stomatitis virus G glycoprotein could be chosen as the envelope allowing a broad host range of transduction. Functionality of particles was monitored by expression of the green fluorescent protein in transduced 3T3 cell clones. This is the first description of a single chimeric virus encoding and releasing functional retroviral vectors, providing proof of principle of the new concept. No replication-competent retrovirus was detectable by sensitive reverse transcriptase assays. Since vaccinia virus has a broad host range, is extremely robust, and can be obtained at high titers and safe nonreplicating vaccinia virus strains are available, the hybrid system may open new perspectives for gene delivery. PMID- 12768021 TI - Functional characterization of the nuclear localization signal for a suppressor of posttranscriptional gene silencing. AB - The nucleus-localized C2 protein of Tomato yellow leaf curl virus-China (TYLCV-C) is an active suppressor of posttranscriptional gene silencing (PTGS). Consistently, infection with TYLCV-C resulted in PTGS arrest in plants. The C2 protein possesses a functional, arginine-rich nuclear localization signal within the basic amino acid-rich region (17)KVQHRIAKKTTRRRR(31). When expressed from potato virus X, C2-RRRR(31)DVGG (in which the four consecutive arginine residues (28)RRRR(31) were replaced with DVGG) that had been tagged with a green fluorescent protein (GFP) failed to transport GFP into nuclei and was dysfunctional in inducing necrosis and suppressing PTGS in plants. Amino acid substitution mutants C2-K(17)D-GFP, C2-HR(21)DV-GFP, and C2-KK(25)DI-GFP localized to nuclei and produced necrosis, but only C2-K(17)D-GFP suppressed PTGS. The N-terminal portions C2(1-31) and C2(17-31) fused in frame to GFP were capable of targeting GFP to nuclei, but neither caused necrosis nor affected PTGS. Our data establish that nuclear localization is likely required for C2 protein to function in C2-mediated induction of necrosis and suppression of PTGS, which may follow diverse pathways in plants. Possible mechanisms of how the C2 protein involves these biological functions are discussed. PMID- 12768023 TI - Interactions of rotavirus VP4 spike protein with the endosomal protein Rab5 and the prenylated Rab acceptor PRA1. AB - Rotavirus spike protein VP4 is implicated in several important functions, such as cell attachment, penetration, hemagglutination, neutralization, virulence, and host range. It is present at the plasma membrane and colocalizes with the cytoskeleton in infected cells. We looked for cellular partners responsible for the localization of VP4 by two-hybrid screening of a monkey CV1 cell cDNA library. In the screen we isolated repeatedly three cDNAs encoding either two isoforms (a and c) of Rab5 protein or the prenylated Rab acceptor (PRA1). The small GTPase Rab5 is a molecule regulating the vesicular traffic and the motility of early endosomes along microtubules. Rab5 interacts with a large number of effectors, in particular with PRA1. Interactions of VP4 with both partners, Rab5 and PRA1, were confirmed by coimmunoprecipitation from infected- or transfected cell lysates. Interaction of Rab5 and PRA1 was restricted to free VP4, since neither triple-layered particles nor NSP4-VP4-VP7 heterotrimeric complexes could be coprecipitated. Site-directed and deletion mutants of VP4 were used to map a VP4 domain(s) interacting with Rab5 or PRA1. Of the 10 mutants tested, 2 interacted exclusively with a single partner. In contrast, the domain extending from amino acids 560 to 722 of VP4 is essential for both interactions. These results suggest that Rab5 and PRA1 may be involved in the localization and trafficking of VP4 in infected cells. PMID- 12768022 TI - Intraventricular brain injection of adeno-associated virus type 1 (AAV1) in neonatal mice results in complementary patterns of neuronal transduction to AAV2 and total long-term correction of storage lesions in the brains of beta glucuronidase-deficient mice. AB - Inherited metabolic disorders that affect the central nervous system typically result in pathology throughout the brain; thus, gene therapy strategies need to achieve widespread delivery. We previously found that although intraventricular injection of the neonatal mouse brain with adeno-associated virus serotype 2 (AAV2) results in dispersed gene delivery, many brain structures were poorly transduced. This limitation may be overcome by using different AAV serotypes because the capsid proteins use different cellular receptors for entry, which may allow enhanced global targeting of the brain. We tested this with AAV1 and AAV5 vectors. AAV5 showed very limited brain transduction after neonatal injection, even though it has different transduction patterns than AAV2 in adult brain injections. In contrast, AAV1 vectors, which have not been tested in the brain, showed robust widespread transduction. Complementary patterns of transduction between AAV1 and AAV2 were established and maintained in the adult brain after neonatal injection. In the majority of structures, AAV1 transduced many more cells than AAV2. Both vectors transduced mostly neurons, indicating that differential expression of receptors on the surfaces of neurons occurs in the developing brain. The number of cells positive for a vector-encoded secreted enzyme (beta-glucuronidase) was notably greater and more widespread in AAV1 injected brains. A comprehensive analysis of AAV1-treated brains from beta glucuronidase-deficient mice (mucopolysaccharidosis type VII) showed complete reversal of pathology in all areas of the brain for at least 1 year, demonstrating that the combination of this serotype and experimental strategy is therapeutically effective for treating global neurometabolic disorders. PMID- 12768024 TI - Prime-boost immunization schedules based on influenza virus and vaccinia virus vectors potentiate cellular immune responses against human immunodeficiency virus Env protein systemically and in the genitorectal draining lymph nodes. AB - Vaccines that elicit systemic and mucosal immune responses should be the choice to control human immunodeficiency virus (HIV) infections. We have previously shown that prime-boost immunizations with influenza virus Env and vaccinia virus (VV) WR Env recombinants induced an enhanced systemic CD8(+) T-cell response against HIV-1 Env antigen. In this report, we analyzed in BALB/c mice after priming with influenza virus Env the ability of two VV recombinants expressing HIV-1 Env B (VV WR Env and the highly attenuated modified VV Ankara [MVA] Env) to boost cellular immune responses in the spleen and in the lymph nodes draining the genital and rectal tracts. Groups of mice were primed by the intranasal route with 10(4) PFU of influenza virus Env and boosted 14 days later by the intraperitoneal or intranasal route with 10(7) PFU of MVA Env or VV WR Env, while the control group received two immunizations with influenza virus Env. We found that the combined immunization (Flu/VV) increased more than 60 times the number of gamma interferon-specific CD8(+) T cells compared to the Flu/Flu scheme. Significantly, boosting with MVA Env by the intraperitoneal route induced a response 1.25 or 2.5 times (spleen or genital lymph nodes) higher with respect to that found after the boost with VV WR Env. Mice with an enhanced CD8(+) T-cell response also had an increased Th1/Th2 ratio, evaluated by the cytokine pattern secreted following in vitro restimulation with gp160 protein and by the specific immunoglobulin G2a (IgG2a)/IgG1 ratio in serum. By the intranasal route recombinant WR Env booster gave a more efficient immune response (10 and 1.3 times in spleen and genital lymph nodes, respectively) than recombinant MVA Env. However, the scheme influenza virus Env/MVA Env increased four times the response in the spleen, giving a low but significant response in the genital lymph nodes compared with a single intranasal immunization with MVA Env. These results demonstrate that the combination Flu/MVA in prime-booster immunization regimens is an effective vaccination approach to generate cellular immune responses to HIV antigens at sites critical for protective responses. PMID- 12768025 TI - Charged assembly helix motif in murine leukemia virus capsid: an important region for virus assembly and particle size determination. AB - We have identified a region near the C terminus of capsid (CA) of murine leukemia virus (MLV) that contains many charged residues. This motif is conserved in various lengths in most MLV-like viruses. One exception is that spleen necrosis virus (SNV) does not contain a well-defined domain of charged residues. When 33 amino acids of the MLV motif were deleted to mimic SNV CA, the resulting mutant produced drastically reduced amounts of virions and the virions were noninfectious. Furthermore, these viruses had abnormal sizes, often contained punctate structures resembling those in the cell cytoplasm, and packaged both ribosomal and viral RNA. When 11 or 15 amino acids were deleted to modify the MLV CA to resemble those from other gammaretroviruses, the deletion mutants produced virions at levels comparable to those of the wild-type virus and were able to complete one round of virus replication without detectable defects. We generated 10 more mutants that displayed either the wild-type or mutant phenotype. The distribution of the wild-type or mutant phenotype did not directly correlate with the number of amino acids deleted, suggesting that the function of the motif is determined not simply by its length but also by its structure. Structural modeling of the wild-type and mutant proteins suggested that this region forms alpha-helices; thus, we termed this motif the "charged assembly helix." This is the first description of the charged assembly helix motif in MLV CA and demonstration of its role in virus budding and assembly. PMID- 12768026 TI - Progressive truncations C terminal to the membrane-spanning domain of simian immunodeficiency virus Env reduce fusogenicity and increase concentration dependence of Env for fusion. AB - The simian immunodeficiency virus (SIV) transmembrane (TM) protein, gp41, has multiple functions, which include anchoring the glycoprotein complex in the lipid envelope of the virus and mediating fusion of the virus and host cell membranes. Recently, a series of mutants of the SIVmac239 TM protein that have truncations at the carboxyl terminus of the membrane-spanning domain (MSD) have been characterized (J. T. West, P. Johnston, S. R. Dubay, and E. Hunter, J. Virol. 75:9601-9612, 2001). These mutants retained membrane anchorage but demonstrated reduced fusogenicity and infectivity as the MSD length was shortened. We have established a novel three-color fluorescence assay, which allows qualitative confocal and quantitative flow cytometric analyses, to further characterize the nature of the fusion defect in five of the MSD mutants: TM185, TM186, TM187, TM188, and TM189. Our analysis showed that each mutant could mediate complete lipid and aqueous dye transfer at early time points after effector and target cell mixing. No hemifusion with only lipid dye flux was detected. However, another intermediate fusion stage, which appears to involve small-fusion-pore formation that allowed small aqueous dye transfer but prevented the exchange of large cytoplasmic components, was identified infrequently in mutant-Env expressing cell and target cell mixtures. Quantitative flow cytometric analysis of these mutants demonstrated that the TM187, TM188, and TM189 mutants were significantly more fusogenic than TM185 and TM186 but remained significantly impaired compared to the wild type. Moreover, fusion efficiency showed an increased dependence on the expression level of glycoproteins, suggesting that, for these mutants, formation of an active fusion complex was an increasingly stochastic event. PMID- 12768027 TI - Basic residues of the helix six domain of influenza virus M1 involved in nuclear translocation of M1 can be replaced by PTAP and YPDL late assembly domain motifs. AB - Influenza type A virus matrix (M1) protein possesses multiple functional motifs in the helix 6 (H6) domain (amino acids 91 to 105), including nuclear localization signal (NLS) (101-RKLKR-105) involved in translocating M1 from the cytoplasm into the nucleus. To determine the role of the NLS motif in the influenza virus life cycle, we mutated these and the neighboring sequences by site-directed mutagenesis, and influenza virus mutants were generated by reverse genetics. Our results show that infectious viruses were rescued by reverse genetics from all single alanine mutations of amino acids in the H6 domain and the neighboring region except in three positions (K104A and R105A within the NLS motif and E106A in loop 6 outside the NLS motif). Among the rescued mutant viruses, R101A and R105K exhibited reduced growth and small-plaque morphology, and all other mutant viruses showed the wild-type phenotype. On the other hand, three single mutations (K104A, K105A, and E106A) and three double mutations (R101A/K102A, K104A/K105A, and K102A/R105A) failed to generate infectious virus. Deletion (Delta YRKL) or mutation (4A) of YRKL also abolished generation of infectious virus. However, replacement of the YRKL motif with PTAP or YPDL as well as insertion of PTAP after 4A mutation yielded infectious viruses with the wild-type phenotype. Furthermore, mutant M1 proteins (R101A/K102A, Delta YRKL, 4A, PTAP, 4A+PTAP, and YPDL) when expressed alone from cloned cDNAs were only cytoplasmic, whereas the wild-type M1 expressed alone was both nuclear and cytoplasmic as expected. These results show that the nuclear translocation function provided by the positively charged residues within the NLS motif does not play a critical role in influenza virus replication. Furthermore, these sequences of H6 domain can be replaced by late (L) domain motifs and therefore may provide a function similar to that of the L domains of other negative-strand RNA and retroviruses. PMID- 12768028 TI - A Domain in the C-terminal region of latency-associated nuclear antigen 1 of Kaposi's sarcoma-associated Herpesvirus affects transcriptional activation and binding to nuclear heterochromatin. AB - The latency-associated nuclear antigen 1 (LANA-1) of Kaposi's sarcoma-associated herpesvirus (KSHV) is required for the maintenance and replication of viral episomal DNA. The binding sites for nuclear heterochromatin and transcriptional repressor complexes are located in an amino-terminal region of LANA-1, whereas those for viral episomal DNA, p53, pRB, and members of the BRD/fsh family of nuclear proteins are located in its carboxy-terminal domain. LANA-1 activates or represses several cellular and viral promoters. In this report we show that a domain of 15 amino acids (amino acids 1129 to 1143), located close to the carboxy terminal end of LANA-1, is required for the interaction of LANA-1 with nuclear heterochromatin or nuclear matrix, and for the ability of LANA-1 to activate the Epstein-Barr virus Cp promoter. LANA-1 proteins that are tightly associated with nuclear heterochromatin or matrix differ in molecular weight from LANA-1 proteins that can be dissociated from the nuclear matrix by high-salt buffers, suggesting that posttranslational modifications may determine the association of LANA-1 with nuclear heterochromatin or matrix. PMID- 12768029 TI - Promyelocytic leukemia protein mediates interferon-based anti-herpes simplex virus 1 effects. AB - Herpes simplex virus (HSV) 1 disaggregates the nuclear domain 10 (ND10) nuclear structures and disperses its organizing promyelocytic leukemia protein (PML). An earlier report showed that ectopic overexpression of PML precludes the disaggregation of ND10 but has no effect on viral replication. PML has been reported to mediate the effects of interferon (IFN) and viral mutants lacking ICP0 (Delta alpha 0 mutants). To test the hypothesis that HSV disaggregates ND10 structures and disperses PML to preclude IFN-mediated antiviral effects, we tested the accumulation of viral proteins and virus yields from murine PML(+/+) and PML(-/-) cells mock treated or exposed to IFN-alpha, IFN-gamma, or both and infected with the wild-type or Delta alpha 0 mutant virus. We report the following results. (i) The levels of growth of wild-type and mutant viruses and of accumulation of viral proteins were not significantly different in untreated PML(+/+) and PML(-/-) cells. (ii) Major effects of IFN-alpha and -gamma were observed in PML(+/+) cells infected with the Delta alpha 0 mutant virus, and more minor effects were observed in cells infected with the wild-type virus. The effects of the IFNs on either wild-type or the mutant virus in PML(-/-) cells were minimal. (iii) The mixture of IFN-alpha and -gamma was more effective than either IFN alone, but again, the effect was more drastic in PML(+/+) cells than in PML(-/-) cells. We concluded that the anti-HSV state induced by exogenous IFN is mediated by PML and that the virus targets the ND10 structures and disseminates PML in order to preclude the establishment of the antiviral state induced by IFNs. PMID- 12768031 TI - Impairment of interferon-induced IRF-7 gene expression due to inhibition of ISGF3 formation by trichostatin A. AB - Two members of the signal transducer and activator of transcription family, STAT1 and STAT2, form, together with interferon regulatory factor 9 (IRF-9), the ISGF3 complex that activates the expression of the interferon-stimulated genes (ISG). The ISGF3 complex also participates in the virus-induced alpha/beta interferon (IFN-alpha/beta) gene amplification cascade by up-regulating IRF-7 gene expression. Here, we show that treatment of cells with trichostatin A (TSA), a deacetylase inhibitor, inhibits the virus-induced activation of IFN-alpha/beta promoters and dramatically reduces the ability of different ISG promoters to respond to IFN stimulation. Impairment of IFN-alpha/beta and ISG expression by TSA in infected cells is due to the blockage of interferon-stimulated ISGF3 complex formation, which leads to the abolition of IRF-7 gene expression. We also show that the TSA-dependent inhibition of ISGF3 is related to impaired nuclear accumulation of STAT2. Our data suggest that an acetylation/deacetylation mechanism participates in the regulation of cellular distribution and function of STAT2 in IFN-alpha/beta signaling. PMID- 12768032 TI - Comparison of human immunodeficiency virus type 1 viral loads in Kenyan women, men, and infants during primary and early infection. AB - Steady-state levels of human immunodeficiency virus type 1 (HIV-1) RNA in plasma reached at approximately 4 months postinfection are highly predictive of disease progression. Several studies have investigated viral levels in adults or infants during primary and early infection. However, no studies have directly compared these groups. We compared differences in peak and set point plasma HIV-1 RNA viral loads among antiretrovirus-naive Kenyan infants and adults for whom the timing of infection was well defined. Peak and set point viral loads were significantly higher in infants than in adults. We did not observe any gender specific differences in viral set point in either adults or infants. However, infants who acquired HIV-1 in the first 2 months of life, either in utero, intrapartum, or through early breast milk transmission, had significantly higher set point HIV-1 RNA levels than infants who were infected after 2 months of age through late breast milk transmission or adults who were infected through heterosexual transmission. PMID- 12768030 TI - Activated Notch1 inhibits p53-induced apoptosis and sustains transformation by human papillomavirus type 16 E6 and E7 oncogenes through a PI3K-PKB/Akt-dependent pathway. AB - Activated Notch1 (AcN1) alleles cooperate with oncogenes from DNA tumor viruses in transformation of epithelial cells. AcN1 signaling has pleiotropic effects, and suggested oncogenic roles include driving proliferation through cyclin D1 or the generation of resistance to apoptosis on matrix withdrawal through a phosphatidylinositol 3-kinase (PI3K)-PKB/Akt-dependent pathway. Here, we extend the antiapoptotic role for AcN1 by showing inhibition of p53-induced apoptosis and transactivation. Chemical inhibitors of the PI3K pathway block AcN1-induced inhibition of p53-dependent apoptosis and nuclear localization of Hdm2. We show that expression of wild-type p53 does not inhibit synergistic transformation by AcN1 and human papillomavirus E6 and E7 oncogenes. We suggest that activation of Notch signaling may serve as an additional mechanism to inhibit wild-type p53 function in papillomavirus-associated neoplasia. PMID- 12768033 TI - Simian virus 5 is a poor inducer of chemokine secretion from human lung epithelial cells: identification of viral mutants that activate interleukin-8 secretion by distinct mechanisms. AB - We have compared chemokine secretion from human lung A549 cells infected with simian virus 5 (SV5) with other members of the Rubulavirus genus of paramyxoviruses. High levels of the chemokines interleukin-8 (IL-8) and macrophage chemoattractant protein-1 (MCP-1) were secreted from A549 cells infected with Human parainfluenza virus type 2 (HPIV-2) but not from cells infected with wild-type (WT) SV5. The lack of IL-8 secretion from SV5-infected cells was not due to a global block in all signal transduction pathways leading to IL-8 secretion, since SV5-infected A549 cells secreted IL-8 after stimulation with exogenously added tumor necrosis factor alpha or by coinfection with HPIV-2. A previously described, recombinant SV5 containing substitutions in the shared region of the P/V gene (rSV5-P/V-CPI-) induced IL-8 secretion by a mechanism that was dependent on viral gene expression. By contrast, an SV5 variant isolated from persistently infected cells (Wake Forest strain of Canine parainfluenza virus) induced IL-8 secretion by a mechanism that was largely not affected by inhibitors of viral gene expression. Together, these data demonstrate that SV5 is unusual compared to other closely related paramyxoviruses, since SV5 is a very poor inducer of the cytokines IL-8 and MCP-1. The isolation of two recombinant SV5 mutants that are defective in preventing chemokine induction will allow an identification of mechanisms utilized by WT SV5 to avoid activation of host cell innate immune responses to infection. PMID- 12768035 TI - Cross-reactivity of HLA-B*1801-restricted T-lymphocyte clones with target cells expressing variants of the human cytomegalovirus 72kDa-IE1 protein. AB - The impact of natural polymorphism in a cytomegalovirus-dominant HLA-B(*)1801 restricted epitope, IE1(199-206), on the specific responses of T-cell clones was assessed by measuring their cytolytic activity against target cells expressing mutated recombinant IE1 proteins. Our results suggest an in vivo selection of T lymphocytes that cross-react with multiple IE1 variants. PMID- 12768034 TI - Mutagenesis versus inhibition in the efficiency of extinction of foot-and-mouth disease virus. AB - RNA viruses replicate near the error threshold for maintenance of genetic information, and an increase in mutation frequency during replication may drive RNA viruses to extinction in a process termed lethal mutagenesis. This report addresses the efficiency of extinction (versus escape from extinction) of foot and-mouth disease virus (FMDV) by combinations of the mutagenic base analog 5 fluorouracil (FU) and the antiviral inhibitors guanidine hydrochloride (G) and heparin (H). Selection of G- or H-resistant, extinction-escape mutants occurred with low-fitness virus only in the absence of FU and with high-fitness virus with some mutagen-inhibitor combinations tested. The combination of FU, G, and H prevented selection of extinction-escape mutants in all cases examined, and extinction of high-fitness FMDV could not be achieved by equivalent inhibitory activity exerted by the nonmutagenic agents. The G-resistant phenotype was mapped in nonstructural protein 2C by introducing the relevant mutations in infectious cDNA clones. Decreases in FMDV infectivity were accompanied by modest decreases in the intracellular and extracellular levels of FMDV RNA, maximal intracellular concentrations of FU triphosphate, and a decrease in the intracellular concentrations of UTP. In addition to indicating a key participation of mutagenesis in virus extinction, the results suggest that picornaviruses provide versatile experimental systems to approach the problem of extinction failure associated with inhibitor-escape mutants during treatments based on enhanced mutagenesis. PMID- 12768036 TI - Flavivirus capsid is a dimeric alpha-helical protein. AB - The capsid proteins of two flaviviruses, yellow fever virus and dengue virus, were expressed in Escherichia coli and purified to near homogeneity suitable for biochemical characterization and structure determination by nuclear magnetic resonance. The oligomeric properties of the capsid protein in solution were investigated. In the absence of nucleic acid, both proteins were predominantly dimeric in solution. Further analysis of both proteins with far-UV circular dichroism spectroscopy indicated that they were largely alpha-helical. The secondary structure elements of the dengue virus capsid were determined by chemical shift indexing of the sequence-specific backbone resonance assignments. The dengue virus capsid protein devoid of its C-terminal signal sequence was found to be composed of four alpha helices. The longest alpha helix, 20 residues, is located at the C terminus and has an amphipathic character. In contrast, the N terminus was found to be unstructured and could be removed without disrupting the structural integrity of the protein. PMID- 12768038 TI - Genetic interaction between Dobrava and Saaremaa hantaviruses: now or millions of years ago? PMID- 12768037 TI - Trans activity of the norovirus Camberwell proteinase and cleavage of the N terminal protein encoded by ORF1. AB - The virus-encoded proteinase of Camberwell virus, a genogroup 2 norovirus, was synthesized in Escherichia coli. The purified proteinase had correct N and C termini and showed trans activity in cell-free assays. trans activity was also demonstrated in COS cells transfected with constructs encoding either the proteinase or a proteinase-polymerase fusion. The N-terminal protein of ORF1 was cleaved in COS cells, possibly at the site E(194)/S. PMID- 12768039 TI - Chronic arthritis: current perspectives. PMID- 12768040 TI - Spectrum of congenital heart defects associated with Down Syndrome in high consanguineous Omani population. AB - OBJECTIVE: To identify the pattern of congenital heart defects (CHD) in children with Down Syndrome (DS) in an Omani population, and compare it with CHD in children without DS and with historical cohorts from populations with low prevalence of consanguinity. SETTING: Open-access Paediatric Echocardiography Clinic at Sultan Qaboos University Hospital, Muscat, Oman. DESIGN: Prospective study. METHODS: All children with DS referred to the Clinic from 1995-1998 formed the subjects (Group I). Children without DS or other known associations of CHD seen at the clinic during the same period served as controls (Group II). Two dimensional echo-Doppler studies were performed on both groups and the results compared. RESULTS: CHD were detected in 54/90 (60%) children in Group I, compared to 698/2122 (32.9%) in Group II (P<0.001). The common CHD in Group I included secundum atrial septal defect (ASD; 18/54), atrioventricular septal defect (AVSD; 15/54) and ventricular septal defect (VSD; 14/54), and in Group II included ASD (175/698), VSD (175/698), patent ductus arteriosus (123/698), pulmonary stenosis (PS; 76/698) and AVSD (35/698). AVSD was more common (P <0.001) and PS less common (P = 0.03) in Group I. Aortic stenosis, coarctation of aorta, transposition of great arteries and complex heart diseases were not detected. Compared to several studies from populations with low prevalence of consanguineous marriages, our study showed a higher frequency of CHD in DS (P <0.05). CONCLUSION: A high frequency of CHD was documented in DS-children from a population with widely prevalent consanguinity. AVSD was most frequent in DS. An interesting observation was the relative rarity of some CHD in the DS population studied. PMID- 12768041 TI - Tongue-tie: an update. PMID- 12768042 TI - Assessment of iodine deficiency disorders in urban areas of Udaipur District, Rajasthan. AB - The present study was conducted on 6-12 years school going children of urban areas of Udaipur to assess the prevalence of iodine deficiency disorders. The study revealed a goiter prevalence of 8.4 percent and biochemical deficiency of nearly 8 per cent but the median iodine levels of the study population was 20 mcg/dL. Iodometric titration of salt samples collected from the beneficiaries revealed that more than 85 per cent of the salt samples had within or more than the stipulated levels of iodine 15 ppm. It appears that the population is in the transition phase from iodine deficiency to iodine sufficiency due to successful implementation of salt iodisation program. PMID- 12768043 TI - Urinary N-acetyl-beta-D-glucosaminidase activity in type I diabetes mellitus. AB - We measured urinary albumin excretion rate (AER) and N-acetyl-beta-D glucosaminidase (NAG) activity in relation to disease duration, acetylated hemoglobin (HbA1c), hypertension and puberty in 44 children and adolescents with type 1 diabetes mellitus. AER and Urinary NAG activity were significantly higher in the patients compared to controls (AER 19.4 +/-; 35.8 vs 4.7 +/- 4.4, NAG activity 5.6 +/- 0.6U vs. 1.6 +/- 0.2U). Microalbuminuria was present in seven patients (15.9%), all of whom were pubertal. There was no correlation between AER and urinary NAG activity. There was a significant direct correlation between AER and disease duration (P <0.05), HbA1c (P < 0.05), diastolic blood pressure (P <0.05) and puberty (P <0.05). None of the microalbuminuria related variables was significantly correlated with urinary NAG activity. Puberty was an independent factor for elevated urinary NAG activity. This study shows that urinary NAG is elevated in children and adolescents with type 1 diabetes mellitus, but is not associated with AER related factors except for puberty. Urinary NAG activity does not appear to be a useful marker for early detection of diabetic nephropathy in children and adolescents with type 1 diabetes mellitus. PMID- 12768044 TI - Preserved umbilical cord facilitates antenatal diagnosis of spinal muscular atrophy. AB - Spinal Muscular atrophy (SMA) Type I is a fatal autosomal recessive disease caused by homozygous deletion of telometric region of exon 7/8 of the SMN gene. Prenatal diagnosis is feasible and desirable by most families. We report on prenatal diagnosis of SMAI in a family where dried umbilical cord stump from the deceased affected baby was used to confirm the diagnosis. Prenatal diagnosis was provided in the subsequent pregnancy. We emphasize the need for storing DNA from individuals affected with suspected single gene disorders. PMID- 12768045 TI - Fowler-like syndrome with extreme oligohydramnios, growth restriction and without muscular hypoplasia. AB - We report the seventh family of Fowler like syndrome (proliferative vasculopathy and hydrocephaly-hydrencephaly syndrome) and first case from Indian subcontinent. A 35 weeks extremely growth retarded male baby showed enlarged ventricles, thinned out cerebral cortex, diffuse intra-cerebral as well as peri-ventricular calcification, cerebral and corneal vasculopathy, unilateral micro-ophthalmia along with corneal opacity and depressed pulsatile anterior fontanel. This case was different from others concerning association with extreme oligohydramnios (in contrast to polyhydramnios), extreme growth restriction (in contrast to normal growth) and absence of gross muscle hypoplasia. No causative factors like TORCH infection, chromosomal abnormality or positive family history was noted in this case. PMID- 12768046 TI - Infected esophageal duplication cyst simulating empyema. AB - Duplications of the esophagus are the second most common duplication of the gastrointestinal tract. The children with esophageal duplication cyst usually present with respiratory distress or as asymptomatic thoracic mass found on incidental chest x-ray. We report a case of infected esophageal duplication cyst initially confused with empyema in a two years old boy. PMID- 12768047 TI - Tachycardiac storm in infant with WPW syndrome: "rescue" radiofrequency ablation. AB - A two-month-old child having WPW syndrome and orthodromic tachycardia was on treatment with digoxin, flecainide and amiodarone. Despite this, he continued to have severe, very frequent episodes of tachycardia. The left-sided accessory pathway was hence ablated via a patent foramen ovale. PMID- 12768048 TI - Noonan syndrome. PMID- 12768049 TI - Pityriasis rubra pilaris. PMID- 12768050 TI - Juvenile dermatomyositis. PMID- 12768051 TI - New Lact-aid technique. PMID- 12768053 TI - New Lact-aid technique: queries. PMID- 12768055 TI - Parents Evaluation of Developmental Status (PEDS). PMID- 12768057 TI - Polio eradication in India: what is the future? PMID- 12768058 TI - Current concepts in management of acute diarrhea. PMID- 12768059 TI - Micronutrient profile of Indian children and women: summary of available data for iron and vitamin A. PMID- 12768060 TI - The fetal and early life origins of adult disease. PMID- 12768062 TI - The risk of pregnancy in patients with lupus nephritis. AB - Since many women with systemic lupus erythematosus (SLE) are of child-bearing age and have normal fertility, the clinician must often face many problems relating to pregnancy in patients with lupus nephritis. These include the influence of lupus nephritis on fetal outcome, obstetric complications, and the influence of pregnancy on SLE. Studies published in the 1960s underlined the increased fetal and maternal risk and recommended against pregnancy in lupus patients. The prognosis for non-pregnant SLE active patients was also poor, making it difficult to know whether pregnancy altered the prognosis of the disease. Recent prospective studies indicate that the majority of lupus mothers can sustain pregnancy without detrimental effects, providing that the pregnancy is planned during the inactive phase of the disease. Nevertheless the fetal risk, although progressively reduced during the last 40 years, continues to be higher than in pregnancies of healthy women particularly in patients with antiphospholipid antibodies. PMID- 12768063 TI - Angiopoietins: microvascular modulators with potential roles in glomerular pathophysiology. AB - Angiopoietins are a recently discovered family of growth factors which act on endothelial cells via Tie receptors. They are widely expressed and have essential roles in regulating vascular growth, development, maturation and permeability. Disturbances in microvascular regulation play an important part in a number of diseases prominent in the developed world including diabetes, ischemic heart disease and cancer. It is the interplay between angiopoietins and other factors including vascular endothelial growth factor (VEGF) which determines endothelial behavior both in health and in these diseases. Angiopoietin-1 is unique in its ability to reduce endothelial permeability and it antagonises the effects of VEGF in its permeability and angiogenesis-inducing actions. The renal glomerulus constitutes a highly specialized microcirculation in which the permeability characteristics of the capillary wall allow its unique filtration function. Disturbance of this function may cause a reduction in glomerular filtration rate or proteinuria. Understanding of the regulation of the filtration barrier is incomplete but the expression of angiopoietins in the glomerulus suggests a mechanism for maintenance of the glomerular endothelium and modulation of the actions of glomerular VEGF. As has been clearly shown for VEGF, angiopoietins are likely to be involved in glomerular disease and recovery from it. Manipulation of angiopoietins has a wide range of potential therapeutic applications from inhibition of diabetic retinal neovascularisation to promotion of glomerular repair. PMID- 12768064 TI - The bone morphogenetic proteins (BMPs). Their role in renal fibrosis and renal function. AB - Recently, a protein has emerged as a potential renotrophic factor: bone morphogenetic protein-7 (BMP-7). In a preventive protocol, BMP-7 treatment was found to significantly decrease renal injury in a rat model of ureteral obstruction (UUO), when treatment was initiated at the time of injury. Subsequent studies suggested that BMP-7 treatment also attenuated renal fibrosis when administered after renal fibrosis had begun. This treatment protocol was also found to increase significantly renal function from the levels measured in the vehicle-treated group. BMP-7 also partially reversed the diabetic nephropathy induced in rats by a single dose of streptozotocin. It restored glomerular filtration rate (GFR), decreased the excretion of protein, and restored histology towards normal. These studies also highlight the value of histological parameters as indicators of renal function and the potential of renal homeostatic factors in the treatment of kidney disease. PMID- 12768065 TI - Mechanisms of repair after kidney injury. AB - Kidney injury is repaired by inflammatory and non-inflammatory mechanisms, with the extent of recovery based on severity of the insult. Critical to the assessment of kidney repair is the ability to differentiate functional recovery from structural repair: compensatory increases in the function of intact residual nephrons often mask the inability of the kidney to heal or replace damaged structures. The mechanisms of repair reflect three levels of injury, which are handled differently by the kidney. First, DNA damage is countered by proof reading DNA polymerases, backed by other controls for sequence misalignment/nucleotide replacement. If DNA cannot be repaired, cells harboring mutation(s) are lost through apoptosis, which is also critical to the disposal of kidney cells and infiltrating leukocytes in both acute and chronic ischemic, immunological, or chemical damage. This leaves room for a second mechanism of repair, i.e., cellular proliferation. At least 5 types of reparative proliferation are known to occur, some of which involve stem cell differentiation and perhaps immigration from distant reservoirs. The final type of repair is referred to as structural repair, actually quite limited by lack of postnatal nephrogenesis in the human kidney. Certain forms of recovery after acute tubular necrosis involve extensive remodeling of the proximal tubule, where integrity of the basement membrane is required for successful repair. Contrary to the long held belief that only acute injury can be repaired, while ongoing chronic damage leads to progressive nephron loss, evidence is emerging that some degree of renal remodeling occurs even in the presence of persistent structural changes. PMID- 12768066 TI - A re-evaluation of the renal ablation model of progressive renal disease in rats. AB - BACKGROUND: The remnant kidney model, usually involving sudden removal or ablation of 1- 1 / (2) to 1-5 / (6) of renal mass, results in compensatory hypertrophy followed by hypertension, proteinuria and declining glomerular filtration rate (GFR) associated with focal (FSG) and then global glomerulosclerosis (GS) and tubulointerstitial injury (TI). Since most renal diseases involve much more gradual injury, we asked whether slow ablation (SA) produced a different natural history than fast ablation (FA). METHODS: Male Munich-Wistar rats underwent heminephrectomy, 3 weeks later a second, and 3 weeks later a third heminephrectomy (SA). They were compared to littermates undergoing simultaneous removal of 1- 1 / (2) kidneys (FA) and sham operated controls (C). RESULTS: Three weeks after the second heminephrectomy, the SA rats had no FSG and glomerular volume (GV) was similar to that of FA rat renal tissue removed at that time. Eight weeks following the final surgical procedure (FSP), the SA and FA groups had similar blood pressures (BP) but higher than C. Albumin excretion rates (AER) were higher in SA and FA vs. C at 1 month after the FSP and, throughout most of the subsequent 5 months, greater in the SA vs. FA groups. At 24 weeks, cortical interstitial fractional volume was double C values in both the SA and FA groups. Percentage of glomeruli with FSG and size (score) of FSG lesions was much higher in SA and FA than C. Moreover, the percentage of FSG in SA (61.2+/-16%) and FSG score (1.7+/-0.7) was greater than in FA animals (35.6+/ 11.9% and 0.9+/-0.4, p<0.01 for each comparison). Mean GV, increased at 24 weeks in both groups over C (1.4+/-0.2 X 10(6) micro m(3)) was greater in SA (3.4+/-0.7 X 10(6) micro m(3)) than FA rats (2.1+/-0.4 X 10(6) micro m(3); p<0.005). CONCLUSIONS: The gradual uninephrectomy in the SA group, insufficient per se to produce significant renal damage, preconditioned the residual kidney, upon further removal of another 1 / (2) kidney, to more albuminuria and FSG lesions than occurred following sudden 1- 1 / (2) nephrectomy, despite similarly elevated BP. Perhaps more time for glomerular enlargement in the SA group preconditioned the remnant kidney to accelerated injury. PMID- 12768067 TI - Heparin reduces glomerular infiltration and TGF-beta protein expression by macrophages in puromycin glomerulosclerosis. AB - BACKGROUND: In a number of experimental models of nephropathy, heparin is renoprotective because it inhibits mesangial matrix synthesis and cell proliferation; in most of these models, glomerular macrophage infiltration has a pathogenic role. We investigated the hypothesis that heparin might also be renoprotective by modulating the macrophages in various ways in the chronic puromycin glomerulosclerosis model. METHODS: We studied the effect of a 3 month course, two different dosages of a non-anticoagulant heparin by immunohistochemical evaluation of the number of macrophages (ED-1 positive cells) in glomeruli, as well as the expression and deposition of TGF-beta and latent TGF beta binding protein in foam cells and mesangial matrix. RESULTS: The renoprotective effect of heparin in this model was confirmed by our observation of lower urine protein and albumin excretion, and a reduced glomerular sclerosis score. These effects were associated with the prevention of macrophage glomerular infiltration, and the inhibition of the TGF-beta axes in foam cells as shown by the reduction in cytoplasm immunostaining for TGF-beta and LTBP-1; heparin also reduced peri-macrophagic collagen IV deposition. CONCLUSIONS: The inhibition of TGF-beta in macrophages seems to be part of heparin general activities. The inhibitory effect of heparin on macrophage infiltration and TGF-beta synthesis in this renal disease model supports the notion that heparin and derived molecules constitute potentially useful therapeutic agents in nephropathies. PMID- 12768068 TI - Protective effect of quercetin on renal ischemia/reperfusion injury in rats. AB - BACKGROUND: There is increasing evidence to suggest that toxic oxygen radicals play a role in the pathogenesis of ischemia/reperfusion (I/R) injury in the kidney. The aim of this study was to evaluate whether quercetin, an oxygen free radical scavenger, protects kidney tissue. METHODS: A renal I/R injury was induced by a left renal pedicle occlusion by ischemia for 45 min, followed by 60 mins of reperfusion with contralateral nephrectomy in rats. The rats were pretreated intraperitoneally with a quercetin suspension (50 mg/kg) 60 min before the ischemia induction. Thiobarbituric acid reactive substances (TBARS), protein carbonyl content, tumor necrosis factor alpha (TNF-alpha), reduced glutathione (GSH) levels, myeloperoxidase (MPO) catalase (CAT) and superoxide dismutase (SOD) activities were determined in renal tissue. RESULTS: There were 3 groups of rats, the control group, the I/R group and the I/R+Q group. Our results indicate that TBARS, TNF-alpha levels, MPO activity and protein carbonyl content were significantly higher in the I/R group than those in the control group (p<0.05, p<0.01, p<0.01 and p<0.01, respectively). Quercetin administration significantly decreased these parameters (p<0.05, p<0.01, p<0.05 and p<0.01, respectively). GSH levels, SOD, and CAT activities significantly decreased after I/R injury when compared to the control group (p<0.01, p<0.05 and p<0.01, respectively). Quercetin treatment significantly increased GSH levels and activities of these enzymes when compared to the I/R group (p<0.05, p<0.01, p<0.05, respectively). CONCLUSIONS: These results suggest that quercetin reduces renal oxidative injury and facilitates repair. Quercetin can have a role in a renoprotective therapeutic regimen. PMID- 12768069 TI - The effect of oxidative stress inhibition with trimetazidine on the peritoneal alterations induced by hypertonic peritoneal dialysis solution. AB - BACKGROUND: Chronic peritoneal dialysis (PD) can result in several peritoneal alterations of varying degree, which lead to progressive reduction in dialytic efficacy. Although its pathogenesis has not been clarified yet, it has been proposed that high glucose induced oxidative stress generation within the peritoneal membrane plays an important role in leading to membrane alterations. The aim of this study was to investigate the effect of oxidative stress inhibition on peritoneal alterations induced by hypertonic PD solutions in rats. METHODS: The rats were divided into three groups receiving no treatment (the control group), hypertonic PD solution intraperitoneally (ip) only (the hypertonic dextrose group) and hypertonic PD solution ip plus trimetazidine (TMZ) orally (TMZ group). After 4 weeks, a one-hour peritoneal equilibration test (PET) was performed. Dialysate-to-plasma urea ratio (D/P urea), glucose reabsorption (D(1)/D(0) glucose), ultrafiltration volume (UF) and the level of dialysate protein were determined. The levels of malondialdehyde (MDA), vascular endothelial growth factor (VEGF) and the activity of glutathione peroxidase (GPx) were investigated in the peritoneal tissue lysates. The peritoneal membrane was evaluated histologically by light microscopy. RESULTS: Compared to the control group, peritoneal function tests (UF: 3.6 +/- 0.4 vs. 1.2 +/- 0.6 mL, D/P urea: 0.57 +/- 0.03 vs. 0.76 +/- 0.04, D(1)/D(0) glucose: 0.46 +/- 0.02 vs. 0.33 +/- 0.05) and morphology (thickness: 4.4 +/- 0.5 vs. 61 +/- 14 micro m and neovascularisation: 0.2 +/- 0.4 vs 2.4 +/- 0.8 number/field) were dramatically altered in the hypertonic PD solution-treated rats. Likewise, higher levels of VEGF, MDA and decreased activity of GPx were determined in the hypertonic PD solution-treated rats. Although peritoneal thickness (37 +/- 17 micro m) was not completely decreased, peritoneal functions were protected in the TMZ group (UF: 4.0 +/- 0.4 mL, D/P urea: 0.62 +/- 0.06, D(1)/D(0) glucose: 0.43 +/- 0.02). In the TMZ group, MDA and VEGF levels and neoangiogenesis were significantly less than those of the hypertonic dextrose group. In addition, GPx activity significantly increased in the TMZ group. CONCLUSIONS: These data demonstrated that not only generating oxidative stress but also attenuating antioxidative system and high glucose concentration can cause structural and functional alterations within the peritoneal membrane. TMZ can preserve these alterations by inhibiting the oxidative stress within the peritoneal membrane. PMID- 12768070 TI - Lipids, blood pressure and bone metabolism after growth hormone therapy in elderly hemodialysis patients. AB - BACKGROUND: Previously we have demonstrated anabolic effects and improved functional status after growth hormone (GH)-therapy in elderly patients in chronic hemodialysis. The aim of this study is to elucidate the effects of GH therapy on lipid profiles, blood pressure and bone metabolism. METHODS: Twenty patients, mean age 73 years, were randomized into two groups i), growth hormone (rHuGH) therapy at a dose of 0.2 IU/kg/BW, or ii) placebo subcutaneously after each dialysis session in a scheme of 3 dialysis per week during 6 months. Two patients in the GH group died (92 and 79 years old) and 1 patient was transplanted. Ten placebo treated patients and 7 GH treated patients were evaluable. RESULTS: The uremic lipid profile with increased triglycerides (TG), low high density lipoproteins, normal lipo-protein Apo-B and relatively low Apo-E values was changed after GH therapy. An unexpected decrease of TG and an indication of decrease of Apo-E values was noted. This differs from GH-treatment to non-uremic adults. Ambulatory 24-hr blood pressures showed a normal circadian rhythm in all patients (GH:n=7, placebo:n=7) at the start and the end of the study. Bone metabolism was increased in the GH group reflected in significant increases of the osteocalcin and telopeptide of type I collagen values. An indication of increased values of propeptide of type I procollagen did not reach statistical significance. CONCLUSIONS: Our study of GH-therapy to elderly patients on hemodialysis demonstrated decreased triglyceride levels, no effect on 24-hr blood pressure and increased bone metabolism. PMID- 12768071 TI - Atorvastatin and simvastatin in patients on hemodialysis: effects on lipoproteins, C-reactive protein and in vivo oxidized LDL. AB - BACKGROUND: Cardiovascular disease as a result of accelerated atherogenesis is common in patients with end-stage renal disease (ESRD). Dyslipidemia may be a major contributor in this process and can be influenced by lipid-lowering drugs (statins). Moreover, statins may exhibit additional inhibitory effects on the atherogenesis, such as a modulation of the immune system as triggered by oxidatively modified LDL and a reduction of the inflammatory marker C-reactive protein (CRP). METHODS: We evaluated in a single-blind randomized trial of 28 ESRD patients on hemodialysis, the dose-depending effects of both atorvastatin and simvastatin on lipids, lipoproteins, LDL particle heterogeneity, high sensitive-CRP, and markers of in vivo LDL oxidation. RESULTS: Both statin therapies significantly lowered total plasma cholesterol and LDL-cholesterol concentrations to the same extent, whereas reduction in the concentrations of triglyceride-rich particles was less pronounced. Furthermore, statin therapy reduced LDL cholesterol in all LDL subfractions, without altering the overall LDL particle density. After both statins plasma hs-CRP concentrations were not significantly reduced; parameters of in vivo LDL oxidation (plasma ox-LDL concentration and the oxidation level of isolated LDL), were significantly decreased. Autoantibodies against ox-LDL, however, did not change during this trial period. CONCLUSIONS: These results show that atorvastatin and simvastatin exhibit comparable favourable effects on lipid profiles in ESRD. Moreover, the reduction of in vivo oxidatively modified LDL as shown in this ESRD population, may indicate that these statins exhibit favourable effects on oxidative stress in vivo. PMID- 12768073 TI - Assessment of health status in chronic hemodialysis patients. AB - BACKGROUND: We compared three ways of assessing health status in chronic hemodialysis patients: generic questionnaire compared with population norms, disease-specific questionnaire, and open questions. METHODS: Hemodialysis patients (n=83) treated in Geneva canton, Switzerland, answered the Kidney Disease Quality of Life (KDQOL-SF) questionnaire, which combines 12 disease specific scales with the generic Short-Form 36 (SF36) health survey, and open questions about the most disturbing and most positive aspects of having end-stage renal disease. SF36 scores were compared with those of the general population, and generic health scales were correlated with dialysis-specific scales. RESULTS: Hemodialysis patients had significantly lower scores than general population controls on 7 of 8 SF36 scales, especially physical functioning (-1.2 standard deviation (SD) units, p<0.001) and general health (-1.2 SD, p<0.001), but their mental health was similar (-0.2 SD, p=0.13). All 12 KDQOL dialysis-specific scores correlated significantly with the SF36 mental summary score, but only 6 correlated significantly with the SF36 physical summary score. Open comments suggested that dialysis itself is the chief problem confronting dialysis patients, but also that the predicament of end-stage renal disease may have a positive impact on the lives of some patients. CONCLUSIONS: While physical problems are the biggest difference between dialysis patients and controls, disease-specific scales and open comments highlighted the importance of psychosocial and treatment-related problems among hemodialysis patients. Generic and disease-specific questionnaires, and open comments provide different information about the health status of dialysis patients. PMID- 12768072 TI - Inflammation, infection and cardiovascular events in chronic hemodialysis patients: a prospective study. AB - BACKGROUND: Cardiovascular (CV) disease is the leading cause of morbidity and mortality in chronic hemodialysis (HD) patients. Inflammation is a potent risk factor for CV disease in the general population. Recent evidence suggests infection, particularly with agents such as Chlamydia pneumoniae (C.pneumoniae) and Helicobacter pylori (H.pylori), as a source of sustained inflammation. Our study tested the hypothesis that C-reactive protein (CRP) and positive serology for antibodies to C.pneumoniae and H.pylori can be associated with the occurrence of new CV events in chronic HD patients. METHODS: We evaluated 76 chronic HD patients (33 women and 43 men, aged 60.5+/-17.3 years) by measuring baseline CRP levels as well as the titres of antibodies (IgG and IgA) to C.pneumoniae and(IgG) to H.pylori. In addition, risk factors such as hypertension, smoking, diabetes, cholesterol levels and albumin were assessed at baseline. The incidence of new CV events (myocardial infarction and ischemic stroke) was recorded during a 36-month follow-up period. The effect of prognostic factors was evaluated by logistic regression analysis. RESULTS: The incidence of CV events was significantly higher in patients seropositive for C.pneumoniae antibodies than in those seronegative (16.1 vs. 4.3 events/100 patient-years, p=0.017, risk ratio 3.76), whereas it did not differ for H.pylori (12.2 vs. 11.7 events/100 patient-years,p=0.91, risk ratio 1.04). Logistic regression analysis showed C.pneumoniae seropositivity (odds ratio 10.11, p=0.04) and CRP levels (odds ratio 1.78, p=0.03) to be independent predictors of the occurrence of CV events. CONCLUSIONS: CRP levels and C.pneumoniae antibodies, but not H.pylori antibodies, were predictors of CV morbidity in the chronic HD patients studied. PMID- 12768074 TI - Bone mass loss in calcium stone disease: focus on hypercalciuria and metabolic factors. AB - BACKGROUND: Several authors have observed that idiopathic calcium stone formers show a bone mass reduction, which is more evident in those with idiopathic hypercalciuria. The aim of this work was the evaluation of osteopenia and osteoporosis rate in a group of idiopathic calcium stone formers. The influence of hypercalciuria, nutritional factors and anthropometric parameters on bone mass was evaluated in these patients as well. METHODS: We enrolled 196 idiopathic calcium stone formers; 102 males, and 94 females. All subjects underwent a metabolic study. BMC and BMD were evaluated as well as QUS. RESULTS: Males showed greater weight, height, BMI, densitometric values and plasma creatinine, uric acid, urea, sodium, magnesium, GFR and urinary osmolarity than females. Moreover males excreted more uric acid, urea, creatinine, sulphate, phosphate, oxalate and citrate than females. The prevalence of osteopenia and osteoporosis, according to T-score, was 54% and 14% respectively. Hypercalciuria was demonstrated in 21.7% of the patients. Hypercalciuric men showed a higher excretion of urea, phosphate, sulphate and magnesium. CONCLUSIONS: Our results confirm the importance of QUS in the evaluation of stone formers' bone mass. Anthropometric characteristics and dietary habits seem to play a role in bone loss. We did not demonstrate any influence of hypercalciuria on bone mass. Although the pathogenesis of bone loss in stone formers still remains unclear, it can be hypothesized that a slight degree of metabolic acidosis, probably of alimentary origin, may be involved in the reduction of bone mass. PMID- 12768075 TI - Spontaneous bilateral perirenal hematomas in two patients with eclampsia. AB - The purpose of this report is to provide two illustrative cases of spontaneous and bilateral perirenal hematomas that occurred during a pregnancy complicated by eclampsia. The sonographic and computed tomographic findings included bilateral perirenal hematoma with no evidence of an underlying malignancy. Since renal hematomas that occur in association with preeclampsia and eclampsia syndrome are extremely rare, but potentially life-threatening complications, prompt laboratory and radiologic evaluations are essential and can reduce the associated morbidity and mortality. PMID- 12768076 TI - Azathioprine hypersensitivity: report of two cases and review of the literature. AB - Azathioprine (AZA) is a widely-used drug in the treatment of different diseases such as vasculitis, systemic lupus erythematosus, rheumatoid arthritis, inflammatory bowel diseases and in renal transplantation. Side effects of AZA can be classified as toxic, mainly dose related (myelosuppression and hepatotoxicity) and idiosyncratic, mainly dose independent. While the toxic effects are common and well documented, the hypersensitivity reactions are rare and it is not often easy to distinguish them from systemic sepsis or disease recurrence. We report two cases of AZA hypersensitivity occurring in patients with anti-neutrophil cytoplasmic antibodies (ANCA)-associated vasculitis each mimicking a vasculitis relapse or a septic complication of immunosuppression, as well as a review of the literature. PMID- 12768077 TI - Hemodialysis vascular access dysfunction: from clinical stenosis to specific cell types. PMID- 12768078 TI - Nephrogenesis regulated by Wnt signaling. AB - Since the discovery that nephron development depends on inductive signaling, the aim has been to characterize the signals involved. These have remained elusive, but some have been identified in recent years, indicating an important role for the members of the Wnt family of secreted growth and differentiation factors. Wnt 4, which is expressed in the developing nephron during the morphogenetic period, is necessary to form nephrons and Wnt signaling is also sufficient to trigger tubule development experimentally in the separated, uninduced kidney mesenchyme. The expression of several other Wnts during kidney development has also been characterized, establishing these as candidates for involvement in the mediation of mesenchymal-epithelial signaling and ureteric bud growth. The varied role of Wnt signaling in the developing kidney is discussed. PMID- 12768079 TI - Renal disease and mitochondrial genetics. AB - Respiratory chain (RC) deficiencies have long been regarded as neuromuscular diseases mainly originating from mutations in the mitochondrial DNA. Oxidative phosphorylation, i.e. adenosine triphosphate (ATP) synthesis-coupled electron transfer from substrate to oxygen through the RC, does not occur only in the neuromuscular system. Therefore, a RC deficiency can theoretically give rise to any symptom, in any organ or tissue, at any age and with any mode of inheritance, owing to the dual genetic origin of RC enzymes (nuclear DNA and mitochondrial DNA). Mitochondrial diseases can give rise to various syndromes or association, namely, neurologic and neuromuscular diseases, cardiac, renal, hepatic, hematological and endocrin or dermatological presentations. The most frequent renal symptom is proximal tubular dysfunction with a more or less complete de Toni-Debre-Fanconi Syndrome. A few patients have been reported with tubular acidosis, Bartter Syndrome, chronic tubulointerstitial nephritis or nephrotic syndrome. The diagnosis of a RC deficiency is difficult when only renal symptoms are present, but should be easier when another, seemingly unrelated symptom is observed. Metabolic screening for abnormal oxidoreduction status in plasma, including lactate/pyruvate and ketone body molar ratios, can help to identify patients for further investigations. These include the measurement of oxygen consumption by mitochondria and the assessment of mitochondrial respiratory enzyme activities by spectrophotometric studies. Any mode of inheritance can be observed: sporadic, autosomal dominant or recessive, or maternal inheritance. PMID- 12768081 TI - Primary hyperoxaluria: genotype-phenotype correlation. AB - Primary hyperoxaluria type 1 (PH1) is an autosomal recessive disorder caused by a deficiency of alanine-glyoxylate aminotransferase (AGT), which is encoded by a single copy gene (AGXT). Molecular diagnosis was used in conjunction with clinical, biochemical and enzymological data to evaluate genotype-phenotype correlation. Patients can present a severe form of PH1, an adult form and a mild to moderate decrease in renal function. Biochemical diagnosis is made by plasma, urine and dialyzate oxalate and glycolate assays, and by liver AGT activity and pyridoxine responsitivity. Molecular genetic diagnosis can be made using different techniques, for example, the single strand conformation polymorphism technique (SSCP), followed by the sequencing of the 11 AGXT exons. The disease is clinically and genetically classified as highly heterogeneous. Mutant alleles can be recognised in 80- 90% of chromosomes, depending on the techniques used. Mutations in exons 1, 2, 4 and 10 are more frequent in Italian patients. Normalized AGT activity seems to be lower in the severe form than in the adult form. Double heterozygous patients present a lower age at disease onset and they were more frequent in the more severe than in mild severe disease. The 444T>C mutation was more frequent in the severe form, while the opposite was observed for 630G>A. 630G>A mutation homozygotes had a higher AGT residual activity. The presence of allelic heterogeneity of the AGXT could be responsible, to some extent, for the phenotypic heterogeneity in PH1. Homozygous genotypes were more frequent than expected and were associated with a less severe form of the disease. PMID- 12768080 TI - Genetic renal disorders with hypomagnesemia and hypocalciuria. AB - The combination of hypomagnesemia and hypocalciuria is the phenotypic signature of two distinct genetic renal tubular transport disorders: Gitelman's syndrome and autosomal dominant isolated renal magnesium wasting. In the past 5 years the genetic defects underlying these disorders have been elucidated through positional candidate cloning approaches. The defective proteins involved in both diseases are located within the distal convoluted tubule (DCT), a segment of the nephron known to play an important role in active magnesium reabsorption in the nephron. The introduction outlines the magnesium handling in the body in general and, in particular, in the kidney, followed by a detailed discussion of Gitelman's syndrome and isolated renal magnesium wasting, including the clinical and biochemical symptoms, genetic aspects and pathophysiology. PMID- 12768082 TI - Type-IV collagen related diseases. AB - Alport syndrome (ATS) is a progressive inherited glomerulonephritis accounting for 1-2% of all patients who start renal replacement therapy, with an estimated gene frequency of approximately 1 in 5000. ATS is a genetically heterogeneous disease, commonly inherited as an X-linked semi-dominant trait, caused by mutations in COL4A5, on the X-chromosome, and only rarely (less than 10% of cases) caused by the COL4A3 or the COL4A4 gene on chromosome 2q. In the X-linked form females are generally less affected than males, microhematuria being the only sign present throughout life, although approximately 30% can progress to end stage renal disease. It became evident in recent years that mutations in the COL4A3 or the COL4A4 gene can give rise not only to autosomal recessive ATS syndrome, in which males and females are severely affected, but also to an autosomal dominant form, where the clinical progression towards impaired renal function can be very slow and also to benign familial hematuria (BFH) in which renal function is preserved. PMID- 12768083 TI - Inherited forms of IgA nephropathy. AB - Simplex and multiplex families with IgA nephropathy (IgAN) have been reported from several ethnic backgrounds, providing the strongest evidence of a role for genetic factors in pathogenesis of IgAN. From a phenotypic point of view, familial and sporadic IgAN cannot be differentiated, and the main clinical and histological features are similar. Traditionally, the case-control study design was employed to identify associations between particular candidate genes, for example, HLA antigens the uteroglobin gene and IgAN, giving conflicting results. Recently, a different approach, using linkage analysis, was undertaken by geneticists at Yale University. A 10-cM genome-wide screen was performed in 30 multiplex IgAN pedigrees, and one locus was mapped (IGAN-1) on chromosome 6q22 23. Future study will be focused on the identification of the gene underlying IGAN-1. This will enable us to understand the molecular pathogenetic basis of IgAN. PMID- 12768084 TI - [Surgical strategies in severe abdominal infections] AB - Initial treatment of peritonitis is largely standardised (elimination source of infection, debridement and intraoperative lavage) but a major problem is the management policy of patients who are at high risk of further infective complications after the initial operation. Existing prognostic scores based on physiological variables, age and chronic disease (APACHE II), and scores that include intraoperative information about the infection (MPI) unfortunately do not seem to be useful in identifying these patients. Management of severe intra abdominal infections is founded on three main principles: 1) supportive care of patient, 2 timely and appropriate antimicrobial therapy and 3) an operative treatment to aim at control the source of infection (evacuate pus, treat abdominal compartment syndrome) and prevent or treat persistant and recurrent infections. In the patients with severe intrabdominal infection there is a great variance in surgical strategies but four may be distingued: continous postoperative peritoneal lavage, relaparotomy on demand ("wait and see" policy), open drainage (laparostomy) and planned relaparotomy. The continous postoperative peritoneal lavage and relaparotomy on demand do not seem to prevent residual o recurrent intrabdominal infections and are associated with a high mortality. The planned relaparotomy seem to decrease the rate of residual peritoneal infection but has a high complication rate. It may be concluded that the ideal operative approach for patients with severe intra-abdominal infection has not been established yet. However, these techniques to be beneficial must be performed in well-selected patients and performed by a team of dedicated surgeons. PMID- 12768085 TI - [Cryptosporidium infection in immunocompetent and immunocompromised host] AB - Prior to 1980 infections with Cryptosporidium species were considered extremely rare in humans. During the eighties, evidence cumulates that this intracellular protozoan was often responsible of self-limiting diarrheal illness in immunocompetent patients and of a prolonged, life-threatening disease in immunocompromised hosts, especially patients with AIDS. The aim of this paper is to review the present knowledge on Cryptosporidium biology, epidemiology, pathogenesis, diagnosis, therapy and highlight recent studies on the clinical aspects of this infection. PMID- 12768086 TI - [Observations about the treatment of Cytomegalovirus retinitis in patients with HIV infection using Ganciclovir and Foscarnet] AB - Objective: The Authors report their experience of Cytomegalovirus (CMV) retinitis therapy in HIV patients, using Ganciclovir and Foscarnet in monotherapy. They also evaluate the reliability of the Polymerase Chain Reaction (PCR) through the qualitative technique as an index of active disease. Methods: 18 patients suffering from CMV retinitis were treated: Ganciclovir was administered at a dosage of 10 mg/kg b.w./day and Foscarnet at 180 mg/kg b.w./day, both of them for 21 days during the induction phase. During the mantainance phase the former was administered at 5 mg/kg b.w./day and the latter at 90 mg/kg b.w./day for 5 days a week. Results: Both the drugs induced the stabilization or regression of the lesions. There was however a relapse with both therapies. We did not observe a significant difference either in the entity and the duration of the stabilization or in the survival from diagnosis time. Finally the PCR method was not helpful in the diagnosis of CMV retinitis. PMID- 12768087 TI - [The behaviour of plasma triglycerides and cholesterol in HIV positive haemophiliacs] AB - Hypertriglyceridemia has frequently been found both in subjects with AIDS and in asymptomatic HIV-positive ones. In order to evaluate the importance of hyperlipemia as an index of the clinical evolution of HIV infection, the levels of triglycerides, total cholesterol and CD4 lymphocytes were determined over a period of 2 years in 8 haemophiliacs with AIDS, 13 asymptomatic HIV-positive and 45 HIV-negative haemophiliacs attending the Operative Unit of Coagulation Disorders of the University of Pisa. The mean concentration of triglycerides and incidence of hypertriglyceridemia were significantly higher in haemophiliacs with AIDS, compared with HIV-negative subjects (p<0.0001), while the triglycerides values of asymptomatic HIV-positives fell between those of the other groups. Cholesterol levels were lower in HIV-positive haemophiliacs and in those with AIDS compared with HIV-negatives. No correlation was found between triglyceride levels and those of CD4 lymphocytes. PMID- 12768088 TI - [Human and animal brucellosis in Caltagirone (CT) area: epidemiological features] AB - The Authors report an epidemiological investigation about human and animal brucellosis, during 1993-97, in Caltagirone (CT) area. A remarkable increase of the cases of human brucellosis related to the increase of ovine-caprine brucellosis cases because of particular climatic conditions during 1996 (morbility index 61 x 100.000 residents) is reported. Despite the great endeavour of the Veterinary Service of District, the results of prophilaxis of ovine caprine brucellosis are modest. Looking forward to the eradication of the animal brucellosis, it is necessary to intervene on the market of the milk and its products to decrease the human brucellosis. PMID- 12768089 TI - [Association of visceral Leishmaniosis and pulmonary tuberculosis: description of a patient] AB - Leishmaniosis, whether localised or disseminated, is mainly correlated to cell mediated immunodeficiency. Immunodeficient patients are also particularly prone to diseases due to Mycobacterium tuberculosis, in whom either the disseminated form or a localisation of the lungs prevails. We report a rather uncommon association of both pathologies successufully treated with N-methylglucamine antimonium followed by an association of rifampycin, isoniazid and ethambutol. The ethiopathogenetic mechanisms, are described. PMID- 12768090 TI - [The Costantin Von Economo's lethargic encefalitis] AB - Starting from the Constantin Von Economo's description of a "new" disease, the lethargic encephalitis, the author delineates the history of an infectious sleeping sickness that caused epidemics in Europe from 1917 to 1928 and led to create, in Italy, the "Institutes for Encephalitis". PMID- 12768091 TI - [Streptogramins: Educational Network] AB - Abstract not avilable PMID- 12768092 TI - Fluid resuscitation of combat casualties. Conference proceedings. June 2001 and October 2001. PMID- 12768093 TI - Tactical combat casualty care: combining good medicine with good tactics. PMID- 12768094 TI - Passing the torch. PMID- 12768095 TI - Combat fluid resuscitation: introduction and overview of conferences. AB - Fluid resuscitation of injured combatants in a tactical setting has special challenges which standard civilian protocols do not address. Over a period of a year, four conferences on combat fluid resuscitation were held. The purpose of these conferences was to develop a consensus regarding contemporary practice and to identify and energize a research agenda. The conferees made specific recommendations for institution of, and endpoints for, resuscitation in the combat setting. They recommended that combat fluid resuscitation in these settings should be initiated with low volume colloid (for U.S. Forces) and hypertonic saline dextran (for those not constrained by U.S. Food and Drug Administration regulations). A specific research agenda was defined. Since that time, these recommendations have been implemented in many North Atlantic Treaty Organization (NATO) forces and a variety of other activities have served to provide expert support and research focus for the special needs of injured combatants. PMID- 12768096 TI - A profile of combat injury. AB - Traumatic combat injuries differ from those encountered in the civilian setting in terms of epidemiology, mechanism of wounding, pathophysiologic trajectory after injury, and outcome. Except for a few notable exceptions, data sources for combat injuries have historically been inadequate. Although the pathophysiologic process of dying is the same (i.e., dominated by exsanguination and central nervous system injury) in both the civilian and military arenas, combat trauma has unique considerations with regard to acute resuscitation, including (1) the high energy and high lethality of wounding agents; (2) multiple causes of wounding; (3) preponderance of penetrating injury; (4) persistence of threat in tactical settings; (5) austere, resource-constrained environment; and (5) delayed access to definitive care. Recognition of these differences can help bring focus to resuscitation research for combat settings and can serve to foster greater civilian-military collaboration in both basic and transitional research. PMID- 12768097 TI - "Don't just do something, stand there!": to teach or not to teach, that is the question--intravenous fluid resuscitation training for Combat Lifesavers. AB - Good Level I scientific evidence supporting the efficacy (decreased morbidity and mortality) of prehospital fluid administration by civilian Emergency Medical Services personnel is lacking. The efficacy of this procedure in the hands of army Combat Lifesavers is even less well substantiated. The purpose of this article is to look critically at the skill of intravenous fluid administration that is taught to army Combat Lifesavers and to consider whether or not the application of that skill is actually beneficial to the majority of patients who are recipients of this procedure. A method is described to assist medical educators in making decisions as to which skills should be taught to health care providers, and this method is loosely applied in the following discussion about whether Combat Lifesavers should receive training to start and administer intravenous fluids. Good scientific studies, based on valid data, need to be performed to determine the efficacy of intravenous fluid administration and other combat medical skills. PMID- 12768098 TI - Recent developments in European colloid solutions. AB - Coagulopathy resulting from hydroxyethyl starch (HES) administration is well documented for high-molecular-weight, highly substituted HES solutions. These are the only starch-based synthetic colloids approved for volume replacement in the United States. Recent developments in European colloid solutions revolve around designing new starch molecules. By increasing the C2:C6 ratio and decreasing the molar substitution to 0.4, a new HES with a molecular weight of 130,000 was introduced. This solution is without significant influence on coagulation. Actual experimental work is concentrating on modulating the inflammatory response of monocytes and endothelial cells after infusion of colloid solutions. This may help prevent or even correct capillary leak, and the intravascular persistence of infused colloidal might be prolonged. PMID- 12768099 TI - Fluid resuscitation: the target from an analysis of trauma systems and patient survival. AB - Much can be learned from studying the deaths that occur in trauma systems as they have developed. Understanding these deaths and the potential effect of trauma systems on reducing death has major implications for designing clinical trials in fluid resuscitation. The availability of new, exciting information regarding fluid composition and physiologic effects argues for new, better-designed clinical trials. By agreeing on the form of resuscitation trials in the future, we will increase our ability to see clinically significant differences in outcome as we move from animal data to clinical efficacy. PMID- 12768100 TI - Fluid resuscitation in the Canadian Forces. AB - In general, the Canadian Forces follow widely accepted principles of fluid resuscitation. These are simply guidelines for fluid resuscitation, and the Canadian Forces currently do not have an absolute doctrine that the clinician in the field must follow. In addition, these principles are adapted to our field environment and the tactical situation that may present itself. In general, Advanced Trauma Life Support guidelines are followed not only for fluid resuscitation but also for the overall resuscitation of the patient. Ringer's lactate is the primary resuscitation fluid that is used. Emphasis is placed on attempting to control ongoing hemorrhage specifically either with direct pressure, surgical control, or splinting of long bone or pelvic fractures at the earliest possible stage. PMID- 12768101 TI - Fluid resuscitation strategies in the Israeli army. AB - Medical training in the Israel Defense Forces (IDF) is currently based on the principles of the Advanced Trauma Life Support course of the American College of Surgeons termed Military Trauma Life Support. The Advanced Trauma Life Support guidelines provide a systematic standardized approach to the treatment of trauma casualties that has been very successful in civilian trauma. On the battlefield, however, these guidelines have been modified according to the combat environment. The factors that influence these changes are tactical considerations, availability and level of training of medical personal, direct enemy fire, medical equipment limitations, means of transportation of casualties, and the variable transportation time from the front line to the first medical echelon. The basic strategy of the IDF is to bring the military physician or paramedic and airlifted surgical units as close as possible to the front line, to minimize evacuation time. Also, evacuation helicopters that land in the combat zone close to the front line, sometimes under direct fire, usually have a military physician on board. The dilemma of "scoop and run" or "stay and stabilize" in hemorrhagic shock has been solved in the IDF toward early rapid evacuation of casualties to a surgical unit. Immediately after airway and breathing have been secured, if evacuation time is less than 1 hour, the intravenous line and fluid resuscitation is started en route to the medical facility. When evacuation time is longer than 1 hour, an intravenous line is always started before evacuation. In controlled hemorrhagic shock, where the source of bleeding has been controlled and evacuation time is less than 1 hour, fluid resuscitation with lactated Ringer's solution or normal saline is started, to achieve normalization of hemodynamic parameters. When evacuation time exceeds 60 minutes, colloids such as Hemaccel or hydroxyethyl starch are added. In uncontrolled hemorrhagic shock, where internal bleeding has temporarily stopped because of hypotension, vasoconstriction, and thrombus formation, aggressive fluid resuscitation with lactated Ringer's solution to achieve normal hemodynamic parameters is prohibited, because it may induce internal rebleeding and hemodynamic decompensation. When evacuation time exceeds 60 minutes, the use of crystalloids and colloids is indicated. If brain injury is suspected, fluid resuscitation should be aimed toward normalization of hemodynamic parameters. PMID- 12768102 TI - Small-volume fluid resuscitation for the far-forward combat environment: current concepts. AB - Hemorrhage remains the primary cause of death on the battlefield in conventional warfare. With modern combat operations leading to the likelihood of significant time delays in air evacuation of casualties and long transport times, the immediate goals of the Army's Science and Technology Objectives in Resuscitation are to develop limited- or small-volume fluid resuscitation strategies, including permissive hypotension, for the treatment of severe hemorrhage to improve battlefield survival and prevent early and late deleterious sequelae. As an example, the U.S. Army has invested much effort in the evaluation of hypertonic saline dextran (HSD) as a plasma volume expander, at one tenth to one twelfth the volume of conventional crystalloids, in numerous animal models of hemorrhage. These studies have identified HSD as a potentially useful field resuscitation fluid. In addition, preliminary studies have used HSD under hypotensive resuscitation conditions, and it has been administered through intraosseous infusion devices for vascular access. This research suggests that many of the difficulties and concerns associated with fluid resuscitation for treating significant hemorrhage in the field can be overcome. For the military, such observations have important implications toward the development of optimal fluid resuscitation strategies under austere battlefield conditions for stabilization of the combat casualty. PMID- 12768103 TI - Fluid resuscitation in modern combat casualty care: lessons learned from Somalia. AB - The medical issues faced by military medics in the combat environment frequently represent a significant variation from their training and civilian experience. The differences between care delivered by military medics under fire and care rendered by civilian medics are profound. The lessons assimilated from extensive discussion and focused conferences form the basis for the proposed changes in combat prehospital care. These differences revolve around a lack of basic monitoring capability, significant logistical constraints, and prolonged evacuation times. The resuscitation algorithm presented in this article represents a consensus of military and civilian trauma experts. PMID- 12768104 TI - Searching for the optimal resuscitation method: recommendations for the initial fluid resuscitation of combat casualties. AB - Resuscitation can exacerbate cellular injury caused by hemorrhagic shock, and the type of fluid used for resuscitation may play an important role in this injury. Unlike some factors in the treatment of combat casualty, the method of resuscitation is under our control. The prevention of cellular injury through wiser resuscitation strategies would be more advantageous than attempting complex immunomodulation after the damage has already occurred. This article summarizes data from a number of studies to illustrate the differential effects of commonly used resuscitation fluids on cellular injury. Our findings show that resuscitation with hypotonic/isotonic crystalloids, including lactated Ringer's (LR) solution, and artificial colloid solutions, elicit severe immune activation and an up-regulation of cellular injury markers. This effect is not seen with plasma, natural colloids (albumin), and fresh whole blood. Hypertonic fluids cause suppression of neutrophil activation and a milder increase in the expression of cell injury markers compared with isotonic fluids. The effect of various resuscitation fluids on core cellular functions such as gene regulation is also summarized in this article. Finally, because of the uniqueness of combat care, a set of new recommendations for initial fluid resuscitation of combat casualties is proposed. PMID- 12768105 TI - Endpoints for fluid resuscitation in hemorrhagic shock. AB - Vigorous intravenous fluid resuscitation has become widely accepted as the optimum management of hemorrhagic shock in trauma. There is now, however, sufficient evidence for this position to be reviewed. Hypotensive or delayed resuscitation has been postulated as a means by which the mortality associated with treatment can be reduced. It has been suggested that overresuscitation with intravenous fluids may worsen hemorrhage. This article discusses the possible adverse effects of "conventional" resuscitation and examines the evidence to support alternative treatment modalities. PMID- 12768106 TI - A review of studies on the effects of hemorrhagic shock and resuscitation on the coagulation profile. AB - BACKGROUND: This study reports on the results of hemorrhagic shock (HS) plus resuscitation on the coagulation profile in severely injured patients and on the role of fresh frozen plasma (FFP) supplementation in a canine HS model. CLINICAL STUDIES: Primary hemostasis (platelet plug), secondary hemostasis (fibrin clot), and fibrinolysis were assayed in 22 injured patients who received an average of 21 units of packed red blood cells (PRBCs), 16.5 L crystalloid solution, and 1.25 L FFP during operation for control of bleeding. Measurements were obtained during operation and postoperatively at 5, 15, 36, and 84 hours and at 26 days. The platelet count during operation was 113,000/mm3, decreased to a nadir of 76,000/mm3 at 48 hours, and then increased to 563,000/mm2 by convalescence. Platelets were not given with resuscitation. The bleeding times and aggregation studies mirrored platelet levels until convalescence. HS led to significant reductions in fibrinogen (factor I), factor V, and factor VIII that normalized by day 1. The fibrinogen increased to supranormal levels by day 4 through day 26. Clotting times mirrored fibrinogen, factor V, and factor VIII levels. Fibrin split products were normal during operation and increased postoperatively through day 26. The increase in fibrin split products paralleled the increase in fibrinogen. EXPERIMENTAL STUDIES: The role of FFP supplementation in HS resuscitation was tested in two separate studies. After HS, the animals were resuscitated with shed blood and crystalloid; half the animals also received FFP. FFP did not improve the coagulation factors, fibrinogen and factors II, V, VII, and VIII. Thus, routine FFP supplementation for HS in humans was abandoned. This led to unexpected postoperative bleeding that in one instance caused death. Therefore, a second controlled study was used in which the FFP was given not only during blood volume restoration but also for an additional hour during continued ongoing controlled hemorrhage without shock. The second study demonstrated that the FFP prevented the reduction in coagulation factors compared with animals not receiving FFP. Clotting times paralleled coagulation protein levels. FFP is now used to supplement HS resuscitation in humans. CONCLUSION: Resuscitation from hemorrhagic shock can be successfully implemented by restoration of blood loss with blood, crystalloid, and FFP added to maintain coagulation proteins. PMID- 12768107 TI - Physiology of colloid-supplemented resuscitation from shock. PMID- 12768108 TI - Crystalloids and colloids in trauma resuscitation: a brief overview of the current debate. AB - BACKGROUND: Controversy regarding crystalloids or colloids for resuscitation has existed for over five decades, and large numbers of clinical trials have failed to resolve the controversy. In fact, the limitations of these studies have intensified the debate. This overview aims to revisit the debate of fluid resuscitation in trauma patients by critically appraising the meta-analyses on the subject. METHODS: This study was a critical analysis of six meta-analyses found by MEDLINE search. RESULTS: Overall, the choice of fluid may have a small or no effect on mortality. In trauma, the use of colloids is associated with a trend toward increased mortality. CONCLUSION: There is an urgent need for well designed clinical trials. Because of many limitations, meta-analysis should be interpreted with caution, possibly as hypothesis generating. However, even considering all weaknesses and nuances of interpretation, the meta-analyses reviewed suggest that trauma patients should continue to be resuscitated with crystalloids. PMID- 12768109 TI - Hypertonic resuscitation: physiologic mechanisms and recommendations for trauma care. AB - Hypertonic saline solutions, with or without added colloid, have received extensive evaluation as volume expanders in both animal studies and clinical trials. Most studies have used 7.5% NaCl/6% dextran 70 (HSD). HSD's primary mechanism of action is rapid osmotic mobilization of cellular water into the blood volume. The measured volume expansion efficiency of HSD is equal to 10 times that of lactated Ringer's solution. Part of HSD's effectiveness is because of the hyperosmotic vasodilation of both systemic and pulmonary vessels. Increased cardiac effectiveness occurs because of the combination of increased preload (venous return) and reduced afterload (vasodilation). Increased cardiac contractility also has been reported in several studies and may play a role, but other studies refute a direct effect on contractility. HSD has been shown to be effective and safe with preexisting dehydration. Animal studies of immune function suggest that increased osmolarity prevents T-cell depression and decreases neutrophil activation. Several perioperative and eight randomized, blinded trauma trials have shown safety and reduced volume needs and suggest increased survival, particularly in head- and penetrating-injury patients. Infusion rates for HSD of 10 to 20 minutes may be recommended for the initial resuscitation of hypotensive trauma. PMID- 12768110 TI - Animal models for hemorrhage and resuscitation research. AB - BACKGROUND: This report summarizes recent workshop discussions on animal models for hemorrhage and resuscitation research, and it is supplemented with relevant current literature. The emphasis is on models for casualty care on the battlefield. RESULTS: Current animal models of hemorrhage and resuscitation vary substantially from one laboratory to another, and are not based on clinical experience. Hemorrhage and resuscitation protocols are arbitrarily determined, and there is no consensus on predictive endpoints. The use of anesthetics in most animal models obscures crucial hemodynamic responses. Environmental variables that influence outcomes by modulating the stress state of the animal are not controlled. A standardized predictive preclinical animal model that addresses these issues is needed. CONCLUSIONS: Recently conscious animal models have been developed that can minimize anesthesia artifacts, and prognostic endpoints have been defined in the clinic. The time may be right to define a predictive animal model for hemorrhage and resuscitation research that will allow new trauma therapies to advance. PMID- 12768111 TI - Blood substitutes and experimental models of trauma. PMID- 12768112 TI - Blood pressure at which rebleeding occurs after resuscitation in swine with aortic injury. AB - BACKGROUND: The appropriateness of vigorous fluid resuscitation to normal blood pressure following hemorrhage in uncontrolled bleeding has recently been questioned due to the possibility of dislodging clots and exacerbating hemorrhage. To develop a rational blood pressure target that maximizes the metabolic benefits of resuscitation without causing increased blood loss, it was first necessary to determine whether there is a reproducible mean arterial pressure (MAP) at which rebleeding occurs. The purpose of this study was to explore the relationship between the rate and time of resuscitation after injury and the rebleeding MAP in an uncontrolled hemorrhage model. METHODS: Sixty-two anesthetized pigs were instrumented with catheters and splenectomized, and suction tubes were placed in the lateral peritoneal recesses to continuously capture shed blood. With the abdomen open, an aortotomy was made in the infrarenal aorta. At either 5, 15, or 30 minutes after the end of the initial hemorrhage, resuscitation with warmed lactated Ringer's solution was begun at either 100 or 300 mL/min. The rebleeding MAP was determined at the moment blood appeared in the suction tubes. RESULTS: The average pressure at the rebleeding point for all animals was MAP = 64 +/- 2, Systolic = 94 +/- 3, and Diastolic = 45 +/- 2 mm Hg. The pressure at which rebleeding occurred in this aortotomy model was not affected by either time of resuscitation (5-30 min), nor was the rebleeding pressure affected by the rate (100 vs. 300 mL/min) of resuscitation. CONCLUSIONS: There was a reproducible pressure at which rebleeding occurred in this model of uncontrolled hemorrhage. The optimal endpoint of resuscitation in patients without definitive hemorrhage control would then be below this rebleeding pressure. PMID- 12768113 TI - Endocrine targets in experimental shock. AB - Modified resuscitation regimens and cytokine blockade/receptor antagonism after trauma have not been successful in decreasing the mortality rates from sepsis in trauma patients; therefore, an alternative approach using endocrine targets as modulators or inhibitors may be useful. Information regarding the influence of gender and hormones on immune and cardiovascular responses after nonthermal trauma-hemorrhagic shock is, on the one hand, considerable but, on the other hand, disappointingly incomplete. Trauma-hemorrhagic shock produces gender dimorphic immune and cardiovascular responses; men exhibit cardiovascular depression and are immunosuppressed, whereas proestrus women do not show cardiovascular or immunologic depression under those conditions. Furthermore, experimental studies have demonstrated the use of hormones, hormone antagonists, sex steroids, and receptor antagonists as salutary adjuncts, without any adverse effects on gastrointestinal, hepatic, and renal functions, for restoring the depressed immune and cardiovascular responses after trauma-hemorrhage. Thus, flutamide, dehydroepiandrosterone, metoclopramide, and 17beta-estradiol, which are readily availably clinically and do not produce any adverse hemodynamic effects, appear to be safe and novel agents/hormones for the treatment of immune and cardiovascular depression after severe blood loss in male and female trauma victims. PMID- 12768114 TI - Endogenous opioid analgesia in hemorrhagic shock. AB - BACKGROUND: Hemorrhagic shock produces an immediate activation of the autonomic nervous system and endogenous opioid pathways. Our studies have demonstrated that endogenous opioid activation aggravates the hemodynamic and inflammatory responses to shock. However, it is unclear whether endogenous opioid activation is triggered by noxious stimuli and furthermore whether it produces analgesia. METHODS: Experiments were conducted in chronically catheterized, conscious, unrestrained, nonheparinized, male, Sprague-Dawley rats subjected to fixed pressure hemorrhage. Blood samples were obtained for determinations of circulating beta-endorphin and substance P. Analgesia was measured using the tail flick response to a noxious stimulus before and during hemorrhage. The contribution of sensory neurons to eliciting the neuroendocrine, opioid, and inflammatory responses to hemorrhage was investigated in capsaicin-treated animals. RESULTS: Hemorrhagic shock produced marked naltrexone-sensitive analgesia without significant modulation of substance P. Peripheral sensory denervation did not alter the hemodynamic, neuroendocrine, or inflammatory responses to shock. CONCLUSION: Endogenous opioid activation during shock produces analgesia. Sensory neuron activation appears to have limited effect on shock-induced hemodynamic and proinflammatory responses. Furthermore, these results suggest that the activation of neuroendocrine and opioid pathways during shock is not likely to be a response to noxious stimuli. PMID- 12768115 TI - Hypertonic saline and the microcirculation. AB - The systemic inflammation that occurs in shock states is believed to promote overexuberant microcirculatory activation, the release of toxic proteases and oxygen radicals causing microvascular damage, and subsequent tissue and organ injury. Although shock-associated microvascular failure is often unresolved after standard resuscitation, hypertonic saline (HTS) appears to reduce microvascular collapse, restoring vital nutritional blood flow. In addition, hypertonic fluids tend to blunt the up-regulation of leukocyte and endothelial adhesion molecules that occurs with isotonic resuscitation of shock. Recently, direct evaluation by intravital microscopy has shown that HTS resuscitation dampens the interactions between leukocytes, platelets, and endothelium found with Ringer's lactate resuscitation. Furthermore, fewer cellular interactions have been correlated with attenuation in microvascular wall permeability after resuscitation with HTS. Better characterization of microcirculatory effects by hypertonic saline may provide mechanisms for improved morbidity and mortality associated with hypertonic resuscitation. PMID- 12768116 TI - Ringer's ethyl pyruvate solution: a novel resuscitation fluid for the treatment of hemorrhagic shock and sepsis. AB - Reactive species of oxygen have been implicated as being important mediators in a variety of pathologic conditions, including burns, various forms of ischemia/reperfusion injury, and hemorrhagic shock. Pyruvate, an intermediate in the metabolism of glucose, is a potent reactive species of oxygen scavenger. Pyruvate, however, is unstable in aqueous solutions, and has not been developed as a therapeutic agent. Ethyl pyruvate, a simple derivative of the parent compound, is thought to be more stable in solution. Ringer's ethyl pyruvate solution (REPS) has been evaluated in a number of preclinical studies using animal models of mesenteric ischemia/reperfusion injury, hemorrhagic shock, and acute endotoxemia. Treatment with REPS, when compared with treatment with Ringer's lactate solution, has been shown to be able to improve survival and decrease expression of proinflammatory mediators. REPS warrants further evaluation for the resuscitation of patients with hemorrhagic shock. PMID- 12768118 TI - Vasopressin deficiency in the syndrome of irreversible shock. PMID- 12768117 TI - Efficacy of hypertonic saline dextran fluid resuscitation for patients with hypotension from penetrating trauma. AB - BACKGROUND: The purpose of this study was to assess whether the administration of hypertonic saline dextran (HSD) was detrimental when administered to patients who were hypotensive because of penetrating injuries to the torso. The administration of HSD causes an immediate and sustained increase in blood pressure that could contribute to an increase in bleeding in the presence of uncontrolled hemorrhage. We prospectively designed a series of questions to be addressed by a meta analysis of individual patient data using a computerized data file and case report forms from a multicenter study of HSD. METHODS: The investigators were "blind" as to the treatment the patient received. Patients (n = 230) with penetrating injuries to the torso were studied as to survival until discharge. The patients were administered 250 mL of HSD or normal saline (standard of care [SOC]) as the initial fluid therapy. RESULTS: Of the 120 patients treated with HSD, 82.5% survived compared with 75.5% for 110 SOC patients (p = 0.19). Sixty eight percent (n = 157) of these patients required surgery. HSD treatment (n = 84) in this population improved survival, 84.5% compared with 67.1% with SOC (n = 73) (p = 0.01). HSD resulted in an increase in blood pressure and a reduction in hematocrit, with no differences noted in fluid requirements or indices of clotting. CONCLUSION: For patients with penetrating injuries to the torso that result in hypotension, initial fluid resuscitation with HSD is beneficial in improving survival, especially if surgery is subsequently required. PMID- 12768119 TI - The use of volume kinetics to optimize fluid therapy. PMID- 12768120 TI - Effects of crystalloid and colloid resuscitation on hemorrhage-induced vascular hyporesponsiveness to norepinephrine in the rat. AB - BACKGROUND: We have shown previously that hemorrhagic hypotension is associated with a progressive development of vascular hyporeactivity to norepinephrine (NE). The present study investigated whether select crystalloid or colloid resuscitation fluids would ameliorate this effect. METHOD: Anesthetized male rats were hemorrhaged to and maintained at a mean arterial pressure (MAP) of 50 mm Hg for 60 minutes. Rats (n = 7 per group) were then resuscitated with lactated Ringer's (LR), 7.5% hypertonic saline (HS) for 1 hour followed by LR (HS-LR), Hespan, or Hextend to restore and maintain MAP to 70 mm Hg over 4 hours. Additional hemorrhaged groups were resuscitated with LR to the baseline MAP (LR BL) or received no resuscitation. A sham hemorrhage group served as controls. The responses of MAP and the blood flow of the superior mesenteric, celiac, left renal, and left femoral arteries to NE (3 microg/kg administered intravenously) were measured at BL (prehemorrhage); at the end of the hypotensive period; and at 1, 2, and 4 hours after resuscitation. RESULTS: Hemorrhagic hypotension significantly (p < 0.01) reduced the NE-induced pressor response in MAP and significantly reduced the contractile responses (reflected by the reduction of blood flow after NE administration) of the four arteries to NE. Hespan and Hextend infusion improved the NE response of MAP and the contractile responses of the observed arteries to NE significantly better than LR, HS-LR, or LR-BL. The colloids improved the vascular contractile responses to NE in the superior mesenteric and left femoral arteries and the pressor response of MAP to NE, to 80% to 90% of their basal response level compared with 40% to 60% with the crystalloid fluids (p < 0.05). LR-BL infusion resulted in hemodilution, with no added benefit to vascular responsiveness. CONCLUSION: These data suggest that hypotensive resuscitation to 70 mm Hg with colloids was better than crystalloids in improving vascular responsiveness to the pressor effects of NE and required smaller volumes. Normotensive resuscitation with LR was not better than hypotensive resuscitation. Not all vasculatures improved equally after fluid resuscitation. PMID- 12768121 TI - Cellular energetics in hemorrhagic shock: restoring adenosine triphosphate to the cells. AB - BACKGROUND: This is a review of studies with two agents, glutamine and crocetin, which have been found to enhance recovery of cellular adenosine triphosphate (ATP) and adenosine diphosphate after hemorrhagic shock. METHODS: The studies used a sublethal (30 minutes) reservoir shock model in 300- to 350-g, male, Sprague-Dawley rats, using either ketamine-xylazine or isoflurane anesthesia. Glutamine was given as a 3% (21 mmol/L) solution in Ringer's lactate (630 mg/kg). Crocetin was given as a 500 nmol/L solution in Ringer's lactate (2 mg/kg). RESULTS: Both glutamine and crocetin caused recovery of ATP to baseline levels (9.0 micromol/g) within 60 to 120 minutes after resuscitation. Xanthine levels returned more rapidly to baseline (0.1 micromol/g). Both agents prevented the elevation in apoptosis seen in controls at 24 and 48 hours. CONCLUSION: Glutamine is a metabolic substrate and a precursor of ATP synthesis. Crocetin enhances oxygen diffusivity in plasma. Both agents restore cellular energy stores to normal after hemorrhagic shock and produce a marked diminution in the extent of apoptosis postshock. Their mechanism of action probably involves prevention of mitochondrial damage. PMID- 12768122 TI - Resuscitating hypothermic dogs after 2 hours of circulatory arrest below 6 degrees C. AB - BACKGROUND: Ultraprofound hypothermia may have a place in trauma rescue and resuscitation. We describe resuscitation of dogs after asanguineous perfusion and circulatory arrest of 2 hours at 2 degrees to 4 degrees C. METHODS: Nine dogs were cooled using a bypass apparatus and their circulating blood replaced with bicarbonated Hextend (Abbott, North Chicago, IL). Perfusion was continued to 2 degrees to 4 degrees C, and 60 mL of 2 mol/L KCl and 20 mL of 50% MgSO(4).7H(2)O were infused intra-arterially, and circulation was arrested for 2 hours. The dogs were then rewarmed, transfused, defibrillated, weaned from bypass, and allowed to awaken. Preoperative and postoperative biochemistry and hematology were compared. RESULTS: Six dogs recovered fully. One of these dogs died of an infection 2 weeks later. Three other dogs never recovered because of technical or procedural difficulties. Biochemical and hematologic parameters were normal by 3 weeks. CONCLUSION: Hypothermic blood substitution with Hextend allows resuscitation after 2 hours of ice-cold circulatory arrest in dogs. PMID- 12768123 TI - Near-infrared spectroscopy-guided closed-loop resuscitation of hemorrhage. AB - BACKGROUND: Endpoint resuscitation has been suggested as a better means to resuscitate penetrating injury. We performed computer-controlled closed-loop resuscitation using invasive cardiac output (CO) or noninvasive skeletal muscle oxygen saturation (SkMusSO(2)) via near infrared spectroscopy (NIRS). METHODS: Conscious sheep received a 4.0-mm aortotomy and uncontrolled hemorrhage at t = 0 min (T0) while resuscitation started at T20 using lactated Ringer's solution. RESULTS: The aortotomy rapidly decreased the mean arterial pressure (MAP) to approximately 30 mm Hg and CO to 20% to 30% of baseline. The SkMusSO(2) endpoint group required only half as much fluid through 4 hours of resuscitation as the CO endpoint group (34.9 +/- 8.4 mL/kg vs. 63.1 +/- 9.4 mL/kg). CO and MAP were lower in the SkMusSO(2) group after T60. Mean infusion volumes were 180% and 100% of the bled volume collected at autopsy in the CO and SkMusSO(2) groups. Brain and muscle oxygenation and base excess were as high or higher in the CO endpoint group. CONCLUSION: Closed-loop resuscitation with either CO or SkMusSO(2) endpoints effectively performs fluid resuscitation of severe uncontrolled hemorrhagic shock. Limited resuscitation may achieve favorable clinical results with volumes less than recommended by Advanced Trauma Life Support guidelines. PMID- 12768124 TI - Oxygen therapeutics in trauma and surgery. AB - A number of oxygen therapeutics have completed safety trials and are now undergoing efficacy evaluation in multicenter phase III trials in North America and Europe. There are numerous potential advantages of these solutions when compared with packed red blood cells. They are readily available and have a long shelf life; do not require type and cross-matching; are virtually free of viral or bacterial contamination; have a much lower viscosity than blood; and may lack the immunosuppressive effects of blood. These products may also deliver more oxygen per unit mass than an equivalent amount of hemoglobin from red blood cells, providing the potential to sustain life in certain clinical situations. A few problems remain, including short biologic half-life, which may limit the application to times when the patient is most acutely anemic (i.e., in the intraoperative or immediate perioperative phase) or for emergent use. Nevertheless, a safe, effective alternative therapy providing oxygen delivery characteristics comparable to red blood cells may soon be available that could have significant impact on the way that we resuscitate trauma patients. PMID- 12768125 TI - Near-infrared spectroscopy in resuscitation. PMID- 12768126 TI - Modeling the two-hit hypothesis for evaluating strategies to prevent organ injury after shock/resuscitation. AB - The major cause of late death in patients sustaining major trauma relates to the development of progressive organ failure. Recent studies suggest that trauma victims are rendered susceptible to the development of organ failure because antecedent shock/resuscitation renders them susceptible to an exaggerated immune response to late inflammatory stimuli, the so-called two-hit hypothesis. In this article, the rationale underlying the use of a "two-hit" model in trauma research is reviewed and a specific example is cited. Importantly, these models permit investigation aimed at understanding the cellular and molecular mechanisms of disease after shock/resuscitation. Interventions such as antioxidant therapy and hypertonic saline resuscitation have a rational basis for use and have been shown to be effective in a rodent two-hit lung injury model. These studies suggest potential use in the critically ill trauma patient population. PMID- 12768127 TI - Burns: military options and tactical solutions. AB - Burn injury remains a constant source of morbidity and mortality in the military environment. The logistic constraints of combat casualty care can make it impossible to provide the large volumes of crystalloid typically used for burn resuscitation. Unlike penetrating trauma, the immediate and sustained fluid requirements necessary for resuscitation of thermal injury preclude the use of limited or hypotensive resuscitation. We examine the physiology, traditional resuscitation strategies, and rationales for the use of novel regimens in the resuscitation of thermal injury. Although strategies such as early use of colloids or hypertonic saline may not reduce morbidity or mortality when compared with large-volume infusions of lactated Ringer's, they can be volume sparing for some hours and sustain life until more definitive therapy is initiated. An intriguing hypothesis is that oral resuscitation can effectively restore plasma volume after thermal injury. We present data from recent experiments of gastric and intestinal infusions of an oral rehydration solution in a porcine burn model that demonstrates restoration of plasma volumes and improvement in hemodynamic parameters associated with significant gastric emptying and intestinal absorption. PMID- 12768128 TI - The tourniquet controversy. PMID- 12768129 TI - Tourniquets for hemorrhage control on the battlefield: a 4-year accumulated experience. AB - BACKGROUND: Tourniquet application is a known means for bleeding prevention in the military prehospital setting. METHODS: This study was a 4-year retrospective analysis of silicone and improvised tourniquet applications by Israeli Defense Force soldiers. RESULTS: Of 550 soldiers who were treated in the prehospital setting, tourniquets were applied to 91 (16%) patients and in less than 15 minutes in 88% of the cases with almost no complications. Penetrating trauma was the main mechanism of injury. The indication was situational and nonmedical in 58 (53%) of the cases. The patients' ischemic time was 83 +/- 52 minutes (range, 1 305 minutes). A total of 78% of applications were effective, with higher success rates for medical staff compared with fellow soldiers and for upper limbs (94%) compared with lower limbs (71%, p < 0.01). CONCLUSION: Tourniquet application is an effective and easily applied (by medical and nonmedical personnel) method for prevention of exsanguination in the military prehospital setting. PMID- 12768130 TI - Crush injury and crush syndrome: a review. AB - Crush injuries and the subsequent systemic manifestations are well described, but there is limited evidence on which to base protocols for optimal management. Given that in most environments such in-juries are relatively rare, there is, however, an undoubted need for such guidelines. This article reviews the currently available evidence and summarizes the treatment options available. PMID- 12768131 TI - Trauma fluid resuscitation in 2010. PMID- 12768132 TI - Malcolm H. Pope: the Fourth ISSLS Wiltse Lifetime Achievement Award sponsored by Stryker recipient. PMID- 12768133 TI - Effects of neutralizing antibodies to tumor necrosis factor-alpha on nucleus pulposus-induced abnormal nociresponses in rat dorsal horn neurons. AB - STUDY DESIGN: The effect of an anti-tumor necrosis factor alpha (anti-TNFalpha) antibody on abnormal discharges caused by application of nucleus pulposus to the nerve root was investigated in an electrophysiologic study. OBJECTIVES: To assess whether inhibition of TNFalpha can reduce nucleus pulposus-induced abnormal discharges. SUMMARY OF BACKGROUND DATA: It has been shown that TNFalpha, a proinflammatory cytokine, is a key pathogenic factor in the development of nucleus pulposus-induced abnormal discharges as a pain sensation. However, the electrophysiologic mechanisms involved in sciatica after disc herniation still have not been elucidated. METHODS: Extracellular activities of wide-dynamic-range neurons were assessed in 21 rats. Autologous nucleus pulposus harvested from the tail was applied to the L5 nerve root. The animals were simultaneously treated with antibodies to TNFalpha (anti-TNF + nucleus pulposus group) and with phosphate-buffered saline (nucleus pulposus group). As a control (control group), a similar volume of muscle was applied to the nerve root with phosphate-buffered saline. Responses of wide-dynamic-range neurons to noxious and innocuous stimuli were examined for 2 hours. RESULTS: Discharges evoked during noxious stimulation and discharges after withdrawal of stimulation in the nucleus pulposus group were significantly higher than those in the control group (P < 0.05). In the anti-TNF + nucleus pulposus group, discharges after withdrawal of stimulation were remarkably inhibited, as compared with those of the nucleus pulposus group (P < 0.05). However, evoked discharges during stimulation apparently were not inhibited. Responses to innocuous stimulation did not change throughout the measurements. CONCLUSIONS: These data indicate that application of TNFalpha antibodies to the nerve root partially prevents the nucleus pulposus-induced abnormal nociresponses. Therefore, anti-TNFalpha treatment may have a therapeutic effect on sciatica after lumbar disc herniation. PMID- 12768134 TI - Effects of immobilization and dynamic compression on intervertebral disc cell gene expression in vivo. AB - STUDY DESIGN: An in vivo analysis of the intervertebral disc's cellular response to dynamic compression and immobilization was performed using a rat-tail model. OBJECTIVE: To assess the effects of immobilization and short-term dynamic compression on intervertebral disc cell expression of anabolic and catabolic genes. SUMMARY OF BACKGROUND DATA: Static compressive loads applied in vivo alter the composition of the disc matrix and cell viability in a dose-dependent manner. The effects of in vivo dynamic compression, which is a more physiologic load, and reported risk factor for low back pain have not been investigated. METHODS: An Ilizarov-type device was implanted on the rat tail and used to determine the effects from 72 hours of immobilization (n = 6), 2 hours of dynamic compression (1 MPa/0.2 Hz) (n = 8), and the coupled effect of immobilization followed by compression (n = 8). Real-time reverse transcription-polymerase chain reaction was used to measure changes in anabolic and catabolic gene levels relative to both internal control subjects and a sham-operated group (n = 7). RESULTS: Immobilization and dynamic compression affect anabolic and catabolic genes, with an overall downregulation of types 1 and 2 collagen and upregulation of aggrecanase, collagenase, and stromelysin in the anulus. The effects of immobilization and compression appear to be additive for collagen types 1 and 2 in the anulus, but not in the nucleus, and not for catabolic genes. CONCLUSIONS: Short-duration dynamic compression and immobilization alter gene expression in the rat disc. In studying the response of the disc to loading, it is necessary to look at both anabolic and catabolic pathways, and to consider strain history. PMID- 12768135 TI - The origin of chondrocytes in the nucleus pulposus and histologic findings associated with the transition of a notochordal nucleus pulposus to a fibrocartilaginous nucleus pulposus in intact rabbit intervertebral discs. AB - STUDY DESIGN: Intact rabbit lumbar intervertebral discs were examined histologically. OBJECTIVES: To demonstrate the origin of chondrocytes in the nucleus pulposus, and to document histologic findings associated with the transition of a notochordal to a fibrocartilaginous nucleus pulposus. SUMMARY OF BACKGROUND DATA: A human nucleus pulposus undergoes a chronological transition from a notochordal to a fibrocartilaginous nucleus pulposus. However, the origin of chondrocytes forming fibrocartilage in the nucleus pulposus and the mechanisms of transition remain unknown. METHODS: Hematoxylin-eosin- and safranin O-stained slides obtained from 125 intact rabbit intervertebral discs were observed with light and polarized light microscopy. RESULTS: Of the 125 intervertebral discs examined, 58 had a notochordal nucleus pulposus. The remaining intervertebral discs had a nucleus pulposus with fibrocartilage lamellas or fibrocartilage fibers. All forms of fibrocartilage lamellas and fibers found in the nucleus pulposus were formed by chondrocytes that had originated and migrated from the cartilage endplate. The origin of chondrocytes proceeded in a centripetal direction from the periphery toward the center of the cartilage endplate. The newly formed fibrocartilage lamellas and fibers, therefore, initially involved replacement of the peripheral regions of the nucleus pulposus, followed by replacement of the central region. This centripetal sequential replacement mechanism decreased the size of the notochordal tissue while increasing the lamellar structure of the intervertebral disc. CONCLUSIONS: Chondrocytes in the intact rabbit nucleus pulposus originated and migrated from the cartilage endplate. The chondrocytes changed notochordal nucleus pulposus into fibrocartilaginous nucleus pulposus by depositing fibrocartilage lamellas and fibers in a centripetal direction. PMID- 12768136 TI - The effect of cement augmentation on the load transfer in an osteoporotic functional spinal unit: finite-element analysis. AB - STUDY DESIGN: The effect of cement augmentation on an osteoporotic lumbar functional spinal unit was investigated using finite-element analysis. OBJECTIVE: To evaluate the influence of cement augmentation on load transfer, stresses, and strains. SUMMARY OF BACKGROUND DATA: Osteoporosis is the most frequent skeletal disease of the elderly, leading to weakness of the bony structures. Cement injection into vertebral bodies has been used to treat osteoporotic compression fractures of the spine. The clinical results are encouraging. Experimental biomechanical studies showed significant increases in stiffness and strength of treated bodies. However, little is known about the consequences for the adjacent, nontreated levels. METHODS: Three-dimensional finite-element models of L2-L3 were developed and the material properties adapted to simulate osteoporosis. The influence of augmentation level as well as uni- and bipedicular filling with polymethylmethacrylate were investigated. Compression, flexion, and lateral bending were simulated. RESULTS: Augmentation increased the pressure in the nucleus pulposus and the deflection of the adjacent endplate. The stresses and strains in the vertebrae next to an augmentation were increased, and their distribution was changed. Larger areas were subjected to higher stresses and strains. The treatment clearly altered the load transfer. Changes to the overall stress and strain distribution were less pronounced for unipedicular augmentation. CONCLUSIONS: Cement augmentation restores the strength of treated vertebrae, but leads to increased endplate bulge and an altered load transfer in adjacent vertebrae. This supports the hypothesis that rigid cement augmentation may facilitate the subsequent collapse of adjacent vertebrae. Further study is required to determine the optimal reinforcement material and filling volume to minimize this effect. PMID- 12768137 TI - Radiologic assessment of interbody fusion using carbon fiber cages. AB - STUDY DESIGN: A comparative study investigated the use of plain static radiographs, flexion-extension radiographs, and thin-section helical computed tomography (CT) scanning in the assessment of anterior lumbar interbody fusion using carbon fiber cages. OBJECTIVE: To compare plain static radiographs, flexion extension radiographs, and thin-section helical computed tomography scans in the assessment of lumbar interbody fusion using carbon fiber cages. SUMMARY OF BACKGROUND DATA: Lumbar interbody fusion has become a popular procedure for the treatment of discogenic back pain. However, there currently is no universally accepted radiologic assessment tool for determining fusion, and the definitive criteria for diagnosing a successful interbody fusion in the lumbar spine remains controversial. METHODS: Plain static radiographs, flexion-extension radiographs, and helical computed tomography scans were performed on 32 patients (49 levels) 5 years after anterior lumbar interbody fusion using carbon fiber cages and autologous bone. A radiologist assessed fusion using the Hutter method to detect movement, whereas a spinal surgeon measured movement in degrees using the Simmons method. Helical computed tomography scans were assessed for the presence of bridging trabecular bone. RESULTS: The fusion rate was 86% on plain radiographs and 84% with the Hutter method. The fusion rate was 74% with the 2 degrees cutoff, and 96% with the 5 degrees cutoff prescribed by the Food and Drug Administration. Fusion on helical computed tomography scans was observed in 65% of the patients. CONCLUSIONS: In the radiologic assessment of interbody fusion using carbon fiber cages, the use of plain radiographs and flexion-extension radiographs produced much higher fusion rates than assessment with thin-section helical computed tomography scans. The thin-section helical computed tomography studies clearly demonstrated the radiographic presence or absence of bridging bone, a property that was not seen with plain static radiographs or flexion extension radiographs. PMID- 12768138 TI - Low back pain after traffic collisions: a population-based cohort study. AB - STUDY DESIGN: A population-based, incidence cohort study was conducted. OBJECTIVE: To measure the incidence and prognosis for collision-related low back pain before and after a change in the insurance compensation system. SUMMARY OF BACKGROUND DATA: Low back pain is a common and costly occupational injury. It also occurs after traffic collisions, but less is known about its frequency and recovery in this setting. METHODS: An incidence cohort of 4473 low back pain injury claims was formed between July 1, 1994 and December 31, 1995 in Saskatchewan. On January 1, 1995 the public insurance system changed from a tort system to a no-fault system, eliminating compensation for pain and suffering. The incidence of claims and the time to claim closure were calculated before and after this change. Prognostic models were built using baseline and follow-up data. RESULTS: The 6-month incidence of claims decreased from 256 to 176 per 100,000 after the insurance change. The median time to claim closure dropped from 505 days for tort claims to 210 days and 216 days for claims made during the first and second 6 months of the no-fault period. Improvements in bodily pain and physical functioning and the absence of depressive symptoms were associated with faster claim closure. High pain intensity, female gender, full-time employment, concentration problems, and lawyer involvement early in the claim process delayed claim closure. CONCLUSIONS: Low back pain is a common traffic injury with a prolonged recovery. Its incidence and prognosis are affected by multiple factors, including the type of compensation system. Our study suggests that biopsychosocial factors are important in determining prognosis. PMID- 12768139 TI - Comparative biomechanical analysis of a cervical cage made of an unsintered hydroxyapatite particle and poly-L-lactide composite in a cadaver model. AB - STUDY DESIGN: A new cage made from a forged composite of unsintered hydroxyapatite particles and poly-L-lactide (F-u-HA/PLLA) is compared biomechanically with the Ray threaded fusion cage. OBJECTIVES: To compare the stability imparted to the human cadaveric spine by two different threaded cervical cages and the effect of cyclic loading on construct stability. SUMMARY OF BACKGROUND DATA: Threaded cages have been developed for use in anterior cervical interbody fusions to provide initial stability during the fusion process. However, metallic instrumentation has several limitations. Recently, totally bioresorbable bone fixation devices made of F-u-HA/PLLA have been developed, including a cage for spinal interbody fusion. However, no biomechanical study has compared the F-u-HA/poly-L-lactide (PLLA) cage with metallic cages. METHODS: For this study, 12 fresh ligamentous human cervical spines (C4-C7) were used. After anterior discectomy across C5-C6, stabilization was achieved with the F-u-HA/PLLA cage in six spines and with the Ray threaded fusion cage in the remaining six spines. Biomechanical testing of the spines was performed with six degrees of freedom before and after stabilization, and after cyclic loading of the stabilized spines (5000 cycles of flexion-extension at 0.5 Nm). RESULTS: The specimens stabilized with either the F-u-HA/PLLA cage or the Ray cage were significantly more stable than the discectomy case in all directions except in extension. In extension, both groups were stiffer, although not at a significant level (P > 0.05). After fatigue, the stiffness, as compared with that in the prefatigue case, decreased in both groups, although not at a significant level. The Ray cage group exhibited better stability than the F-u HA/PLLA cage group in all directions, although a significant difference was found only in right axial rotation. CONCLUSIONS: The F-u-HA/PLLA cage has the possibility to supplant the use of metallic devices in interbody fusions of the cervical spine. PMID- 12768141 TI - Graft migration or displacement after multilevel cervical corpectomy and strut grafting. AB - STUDY DESIGN: A retrospective review of consecutive patients with graft migration or displacement after anterior cervical corpectomy surgery was performed. OBJECTIVES: To examine the associated risk factors and results of treatment among patients who sustained graft displacement or migration after anterior cervical corpectomy surgery. SUMMARY OF BACKGROUND DATA: Graft migration or displacement after anterior cervical corpectomy is a potential complication that may require revision surgery, but because of the low incidence, the factors associated with graft movement and the results of treatment are not well defined. METHODS: All patients who had undergone a cervical corpectomy were examined for graft migration or displacement. None of the patients had a previous cervical laminectomy or prior posterior cervical surgery. All the patients were treated with autogenous strut grafting after decompression. RESULTS: Over a 25-year period, 249 consecutive patients underwent one- to five-level anterior cervical corpectomies and strut grafting. All the patients were fused using autogenous bone grafts (iliac crest or fibula). During the postoperative period, 16 of the patients (10 women and 6 men; average age, 61.4 years) experienced migration of their grafts. The average follow-up period was 4.7 years (range, 2-12 years). The graft migration rates increased with more levels of fusion (odds ratio of 1.65 for having a displaced graft with each additional level): 4 of 95 single-level grafts, 4 of 76 two-level grafts, 7 of 71 three-level grafts, and 1 of 6 for four level grafts. Of the 16 patients with graft migration, 14 had procedures involving a corpectomy of C6 with a fusion inferiorly extending to the C7 vertebral body (P = 0.001, statistically significant difference). Of these 16 patients, 5 underwent revision surgeries acutely for displacement and associated fracture of the inferior graft and vertebral body junction. None of the patients sustained a neurologic or respiratory complication as a result of graft migration ordisplacement. All of the patients went on to successfulfusion. CONCLUSIONS: This study demonstrated that a greater number of vertebral bodies removed and a longer graft are directly related to an increased frequency of graft displacement. Graft displacement may require revision surgery, but no patient in this study experienced a permanent adverse result from this complication. Corpectomies involving a fusion ending at the C7 vertebral body were associated with a higher rate of graft migration. PMID- 12768143 TI - Retrograde ejaculation after anterior lumbar interbody fusion: transperitoneal versus retroperitoneal exposure. AB - STUDY DESIGN: In this multicenter, prospective, 2-year study, 146 male patients underwent a single-level anterior lumbar interbody fusion with a tapered threaded titanium fusion device. All the patients were advised before surgery of the risk for retrograde ejaculation. After surgery, any case of retrograde ejaculation was recorded as an adverse event, and the patient was observed up for the remainder of the study. OBJECTIVE: To determine the incidence of retrograde ejaculation in male patients treated for single-level degenerative lumbar disc disease at L4-L5 or L5-S1 with a stand-alone anterior interbody fusion using tapered, threaded titanium fusion cages. SUMMARY OF BACKGROUND DATA: The incidence of retrograde ejaculation in men after anterior lumbosacral spinal surgery has been reported to range from 0.42% to 5.9%. Various risk factors that increase the chance of retrograde ejaculation have been proposed. METHODS: In this prospective study, 146 male patients underwent an open surgical exposure of the lumbosacral junction and a single-level interbody fusion at either L4-L5 or L5-S1. Assessment of a patient's clinical outcome was based on written questionnaires at 6 weeks and then 3, 6, 12, and 24 months after surgery. Patients were questioned about adverse events at each of these assessments, and any case of retrograde ejaculation was recorded and followed. RESULTS: Retrograde ejaculation developed in 6 of the 146 men after open anterior lumbar interbody fusion surgery. Two cases (1.7%; 2/116) involved patients who underwent a retroperitoneal surgical exposure. Four cases (13.3%; 4/30) involved patients who had a transperitoneal surgical exposure. This difference is statistically significant according to Fisher's exact test (P = 0.017). At 12 months after surgery, 2 patients had resolution of their symptoms: 1 in the retroperitoneal approach group and 1 in the transperitoneal group. At the final 2-year follow-up, no changes in symptoms were reported. One patient in the retroperitoneal approach group (0.86%) and three patients in the transperitoneal group (10%) reported permanent retrograde ejaculation (P = 0.027). CONCLUSIONS: A transperitoneal approach to the lumbar spine at L4-L5 and L5-S1 has a 10 times greater chance of causing retrograde ejaculation in men than a retroperitoneal approach. PMID- 12768144 TI - The natural history of spondylolysis and spondylolisthesis: 45-year follow-up evaluation. AB - STUDY DESIGN: A prospective study of spondylolysis and spondylolisthesis was initiated in 1955 with a radiographic and clinical study of 500 first-grade children. OBJECTIVE: To determine the natural history of spondylolysis and spondylolisthesis. SUMMARY OF BACKGROUND DATA: Most studies on the natural history of spondylolysis and spondylolisthesis are based on patient populations presenting with pain. Critical to any natural history investigation is the study of a population of affected individuals, whether symptomatic or not, from onset of the condition through their lives. METHODS: By study of a population from the age of 6 years to adulthood, 30 individuals were identified to have pars lesions. Data collection at a 45-year follow-up assessment included magnetic resonance imaging, a back pain questionnaire, and the SF-36 Survey. RESULTS: No subject with a pars defect was lost to follow-up evaluation once a lesion was identified. Subjects with unilateral defects never experienced slippage over the course of the study. Progression of spondylolisthesis slowed with each decade. There was no association of slip progression and low back pain. There was no statistically significant difference between the study population SF-36 scores and those of the general population the same age. CONCLUSIONS: This report is the only prospective study to document the natural history of spondylolysis and spondylolisthesis from onset through more than 45 years of life in a population unselected for pain. Subjects with pars defects follow a clinical course similar to that of the general population. There appears to be a marked slowing of slip progression with each decade, and no subject has reached a 40% slip. PMID- 12768146 TI - Sagittal alignment after anterior debridement and fusion with or without additional posterior instrumentation in the treatment of pyogenic and tuberculous spondylodiscitis. AB - STUDY DESIGN: A retrospective clinical study investigated patients undergoing surgery for destructive pyogenic and tuberculous spondylodiscitis. OBJECTIVE: To compare anterior debridement and bone grafting with a combined anterior and posterior procedure in terms of the physiologic alignment of the segmental sagittal spinal profile. SUMMARY OF BACKGROUND DATA: There is considerable agreement in the literature on the indications for surgical treatment of destructive spondylodiscitis. An anterior approach usually is recommended for debridement and bone grafting. Additional posterior instrumentation is applied to reduce kyphotic deformities and to prevent a correction loss. No comparison has been made so far in the literature between repositioning results obtained after surgery for destructive spondylodiscitis and physiologic segmental sagittal angles. METHODS: The surgical results of 49 patients treated by anterior debridement and bone grafting were compared with those of 22 patients who received additional posterior instrumentation. A comparison between the segmental kyphotic angles obtained and the standard values reported in the literature enabled an assessment of the segmental spinal alignment in the sagittal plane. Data were obtained from medical record review, imaging procedures, and patient follow-up examinations. RESULTS: All the subgroups submitted to a combined procedure had a greater preoperative segmental kyphosis angle than those undergoing anterior fusion alone. In marked segmental kyphotic false positioning, good postoperative repositioning was achieved by the combined procedure, and an increase in segmental kyphosis was permanently prevented. CONCLUSIONS: In single level spondylodiscitis with no major substance loss, anterior debridement and bone grafting alone seem to be adequate, especially in the lumbar spine. Additional posterior instrumentation is indicated in multiple-level spondylodiscitis, extensive kyphotic deformity, or both. PMID- 12768147 TI - Transcranial electrical motor-evoked potential monitoring during surgery for spinal deformity: a study of 145 patients. AB - STUDY DESIGN: A descriptive historic cohort study was conducted. OBJECTIVES: To determine intraoperative response amplitude criteria for transcranial electrical motor-evoked potential monitoring that warn of neurologic damage, and to determine the additional value of monitoring six instead of two muscle sites. SUMMARY OF BACKGROUND DATA: Transcranial electrical motor-evoked potential monitoring provides immediate and reliable information about the integrity of the motor pathways during spine surgery. Although this monitoring technique is more frequently used, criteria for interpretation of the amplitude responses have not been defined. METHODS: The intraoperative monitoring outcomes were compared with the patient's clinical outcomes. The sensitivity, specificity, positive predictive value, and negative predictive value were determined for four different monitoring criteria. RESULTS: Transcranial electrical motor-evoked potential monitoring was possible 142 of 145 patients undergoing corrective surgery. In this study, 16 patients had a neurologic event, and 11 patients showed recovery of response amplitude after a second surgical maneuver, whereas the remaining 5 patients had permanent partial neurologic damage. The criterion that at least one of six recordings must show an amplitude decrease of more than 80% was sufficiently strict to achieve a sensitivity of 1.0 and a specificity of 0.91. Less strict criteria, including recording at two instead of six sites, resulted in lower sensitivity, with the lower bound of the 95% confidence interval at 0.62. CONCLUSIONS: Transcranial electrical motor-evoked potential monitoring allowed successful intraoperative monitoring. The criterion of one recording showing a response amplitude decrease of more than 80% during a surgical action can be considered a valuable warning criterion for neurologic damage. The authors also consider that monitoring at six instead of two muscles improves the value of neuromonitoring. PMID- 12768148 TI - The million visual analog scale: its utility for predicting tertiary rehabilitation outcomes. AB - STUDY DESIGN: A longitudinal cohort study involving 1749 patients with chronically disabling spine disorder (CDSD) who underwent tertiary rehabilitation investigated the relation between the Million Visual Analog Scale (MVAS) score and treatment outcome. OBJECTIVES: To determine whether the pretreatment MVAS rating of disability severity is associated with the ability to complete functional restoration rehabilitation, and to determine whether pre- or posttreatment MVAS disability perception is associated with 1-year posttreatment socioeconomic outcomes. The relation of the MVAS to pre- and posttreatment psychosocial measures and physical performance levels also was evaluated. SUMMARY OF BACKGROUND DATA: The MVAS yields a total functional disability score ranging from 0 to 150. Like other "disability inventories," the MVAS differs from a "pain inventory" in that the focus is on disability and function, as opposed to self reported pain. The MVAS may currently be the most powerful functional rating scale because all its questions relate to the patient's ability to perform activities of daily living. It also has the advantage of a visual analog format, which typically is considered more effective than other commonly used self-report formats. METHODS: A large cohort of 1749 patients with CDSD who underwent tertiary rehabilitation was divided into groups by their severity of disability, rated on the MVAS, both at pre- and posttreatment assessment. The patients were divided into groups ranging from "no reported disability" (MVAS = 0) to "extreme disability" (MVAS = 131-150). The distribution into the six groups was assessed on both pre- and posttreatment MVAS ratings. The patients underwent a 3-week functional restoration program consisting of daily quantitatively directed exercise progression and multimodal disability management. Physical capacity and psychosocial assessments, performed before and after treatment, were correlated with the MVAS scores. A 1-year posttreatment clinical interview obtained information on socioeconomic outcomes, which also were correlated with the MVAS ratings. RESULTS: Mantel-Haenszel linear analyses showed a number of relations between demographic variables and both pre- and posttreatment MVAS scores. Most importantly, the findings showed that severe pretreatment MVAS scores were associated with a lower program completion rate (94% vs 89%; P < 0.001) and a higher rate of postrehabilitation health care use from a new provider (12% vs 41%; P < 0.001). Prerehabilitation scores also were linearly related to lower levels of pretreatment physical performance and higher rates of pretreatment depression. More severe posttreatment MVAS scores were associated linearly with a drop in the work return rate from 93% to 63%, a drop in the work retention rate 1 year after rehabilitation from 86% to 44%, and a drop in the financial settlement rate from 94% to 79% (P < 0.001). A linear trend also was found in the rate of postrehabilitation surgeries, with the percentages rising from 0% in the group with no reported disabilities to 12% in the group with extreme disabilities (P < 0.001). CONCLUSIONS: The current study represents the first large-scale examination of the relation between MVAS ratings and treatment outcomes in a CDSD population. These results demonstrate the effectiveness of a simple disability rating scale, such as the MVAS, for systematic disability assessment in potentially predicting treatment outcomes in patients with CDSD. Despite the popularity of other questionnaires, the MVAS is the first disability inventory with demonstrated effectiveness for this purpose in a large CDSD population. PMID- 12768149 TI - Comparison of the Roland-Morris Disability Questionnaire and generic health status measures: a population-based study of workers' compensation back injury claimants. AB - STUDY DESIGN: Workers' compensation back injury claimants completed baseline and follow-up telephone interviews in a prospective population-based cohort study. OBJECTIVE: To compare the Roland-Morris Disability Questionnaire (RDQ) to widely used generic health status measures in a sample of workers with recent work related back injuries in terms of validity, reliability, responsiveness to change, and floor and ceiling effects. SUMMARY OF BACKGROUND DATA: Little research has directly compared the validity and responsiveness of the RDQ to that of the Short-Form 12 or Short-Form 36 health status measures among individuals with back pain. Furthermore, there is little information concerning the validity, reliability, and responsiveness of the RDQ as a measure of functional outcomes for workers with back injuries. METHODS: Approximately 8 weeks (median) after filing low back injury claims, 309 workers completed the RDQ, Short-Form 12, and Short-Form 36 scales and gave information about their work status in computer assisted telephone interviews. An average of 5 months later, 284 workers (91.9%) completed the measures again. RESULTS: The RDQ demonstrated excellent internal consistency and validity through correlations with other measures of physical functioning, ability to discriminate between those working and those not working, and much more responsiveness to change than the Short-Form 12 and Short-Form 36 scales. However, 15% of the sample did not answer one or more RDQ items. CONCLUSIONS: The RDQ is a valid measure of physical disability among workers with back injuries. Its greater responsiveness to change suggests its superiority to the Short-Form 12 and Short-Form 36 as an outcome measure in this population. PMID- 12768151 TI - Trunk muscle strength in flexion, extension, and axial rotation in patients managed with lumbar disc herniation surgery and in healthy control subjects. AB - STUDY DESIGN: A cross-sectional study was conducted. OBJECTIVE: To compare maximal flexion, extension, and rotation strength as well as force-time characteristics of trunk muscles in patients who undergo lumbar disc herniation with those in healthy control subjects 2 months after surgery. SUMMARY OF BACKGROUND DATA: Insufficient attention has been paid to muscle strength characteristics after lumbar disc herniation surgery. METHODS: For this study, 30 postoperative patients with lumbar disc herniation and 30 healthy control subjects volunteered to participate. Isometric trunk flexion, extension in the standing position, and seated rotation strength were tested bilaterally in a neutral posture and at 30 degrees axial prerotation. The area under the curve was calculated to analyze explosive force capacity. Dynamic endurance strength was measured by calculating the repetition maximum. Pain during the strength measurements was assessed by a visual analog scale. RESULTS: The healthy control subjects showed 44% and 36% higher isometric trunk flexion (P < 0.001) and extension (P < 0.001) forces, respectively, than the patients. The respective values for the area under the curve were 41% and 37% higher for the trunk flexors (P < 0.001) and extensors (P < 0.001) in the healthy control subjects than in the patients. The differences in trunk rotation force between the groups were statistically significant when the lower body was rotated 30 degrees to the right (P = 0.023) or to the left (P = 0.043) and the upper body was rotated in the opposite direction. Furthermore, in the dynamic endurance strength test, the healthy control subjects performed 70% more repetitions both for trunk flexors and extensors than did the patients. Some of the patients reported mild pain during the strength measurements, but the level of pain did not correlate with the strength values. CONCLUSIONS: The recovery of maximal endurance and explosive type strength characteristics is incomplete in patients with lumbar disc herniation 2 months after surgery. Active strength training is recommended to restore muscle function in these patients. PMID- 12768152 TI - Lumbar repositioning deficit in a specific low back pain population. AB - STUDY DESIGN: A cross-sectional observational design study was conducted to determine lumbar repositioning error in 15 subjects who had chronic low back pain with a clinical diagnosis of lumbar segmental instability and 15 asymptomatic participants. OBJECTIVE: To determine whether individuals with lumbar segmental instability have a decreased ability to reposition their lumbar spine into a neutral spinal position. SUMMARY OF BACKGROUND DATA: Proprioception of the lumbar spine has been investigated in individuals who have low back pain with variable results. The testing procedure's lack of sensitivity and the nonhomogeneity of groups may be responsible for the conflicting findings. METHODS: Repositioning accuracy of the lumbar spine was assessed using the 3Space Fastrak to determine error in 15 participants with lumbar segmental instability and 15 asymptomatic subjects. The participants were assisted into a neutral spinal sitting posture and then asked to reproduce this position independently over five trials separated by periods of relaxed full lumbar flexion. RESULTS: Lumbosacral repositioning error was significantly greater in participants with lumbar segmental instability than in the asymptomatic group (t[28] = 2.48; P = 0.02. There also was a significant difference between the groups at each individual sensor. CONCLUSIONS: The results of this study indicate that individuals with a clinical diagnosis of lumbar segmental instability demonstrate an inability to reposition the lumbar spine accurately into a neutral spinal posture while seated. This finding provides evidence of a deficiency in lumbar proprioceptive awareness among this population. PMID- 12768155 TI - The case of James A. Garfield: a historical perspective. AB - In 1881, President James A. Garfield was shot in the back and died 79 days later. During this time, many controversies arose that had repercussions for years to come. Who was to manage the President's care? A prominent local physician took on the case, but after Garfield's death, he was highly criticized for inappropriate care and for excluding more highly qualified surgeons. Where was the bullet? Multiple opinions were given including that of Alexander Graham Bell. The correct suggestion turned out to be that of a young, unknown assistant demonstrator of anatomy. What was the proper treatment? Local wound care, removal of the bullet, and laparotomy all were considered. Many have felt that the choice of treatment may have proved to be worse than the injury itself. What did the autopsy show? Even this was controversial, with different observers claiming different results. This historical perspective reviews the case as well the controversies that surrounded it. PMID- 12768153 TI - Reduction of radiculopathy and pain with Oxiplex/SP gel after laminectomy, laminotomy, and discectomy: a pilot clinical study. AB - STUDY DESIGN: Safety using Oxiplex/SP Gel during single-level discectomy for reduction of symptoms associated with unilateral herniation of the lumbar disc was investigated by self-assessment questionnaire and magnetic resonance imaging. OBJECTIVE: To evaluate the safety and assess the efficacy parameters of Oxiplex/SP Gel. SUMMARY OF BACKGROUND DATA: Animal studies demonstrated that Oxiplex/SP Gel (CMC/PEO) reduced epidural fibrosis after lumbar surgery. METHODS: Surgeons examined spine and lower extremities of patients scheduled for discectomy to assess neurologic function and pain. Treated patients received sufficient Oxiplex/SP Gel (1-3 mL) to coat the nerve root and fill the epidural space. The control condition was surgery alone. At baseline, then 30 days, 90 days, and 6 months after surgery, patients completed self-assessment questionnaires concerning leg pain, lower extremity weakness, functional disability, daily living activities, symptoms, and radiculopathy. Magnetic resonance imaging was performed at baseline and 90 days after surgery. At 30 and 90 days after surgery, patients underwent physical examination, wound inspection, and laboratory tests. RESULTS: The surgical procedures were well tolerated by the 23 patients treated with Oxiplex/SP Gel and the 11 control patients. There were no unanticipated adverse events, no clinically significant laboratory results, and no significant differences detected by magnetic resonance imaging. Treated patients had greater reduction in outcome measures at 30 days. The differences in scores were attenuated at 90 days and 6 months. A subgroup, the patients with significant leg pain and weakness at baseline (11 patients treated with Oxiplex/SP Gel and 7 control patients), had greater reduction in outcome measures than the control patients throughout the study. CONCLUSIONS: Oxiplex/SP Gel was easy to use and safe for patients undergoing unilateral discectomy. Greater benefit in clinical outcome measures was seen in gel-treated patients, especially those with severe leg pain and weakness at baseline. PMID- 12768156 TI - Kyphectomy using a surgical threadwire (T-saw) for kyphotic deformity in a child with myelomeningocele. AB - STUDY DESIGN: A 9-year-old boy with severe myelomeningocele kyphosis was treated by kyphectomy using a surgical threadwire. OBJECTIVE: To describe a new method of kyphectomy for severe kyphotic deformity in a child with myelomeningocele using a surgical threadwire. SUMMARY OF BACKGROUND DATA: Although several methods of kyphectomy for severe kyphotic deformity in children with myelomeningocele have been reported, few of these methods allow preservation of the nonfunctioning dural sac and cerebrospinal fluid flow, with the aim of reducing complications. METHODS: The preoperative kyphotic angle was 113 degrees. There was repeated skin ulceration over the apex of the kyphos. Kyphectomy at the Th12 to L3 vertebral levels was performed using a surgical threadwire (T-saw, developed by Tomita and colleagues in 1996), preserving the entire dural sac. RESULTS: The T-saw allowed anterior dissection of the dural sac over the length of the planned resection, thus preserving cerebrospinal fluid flow throughout the entire subarachnoid space. The kyphotic angle was decreased to 10 degrees after the operation, and the postoperative clinical course was uneventful. At the 2-year follow-up assessment, the kyphotic angle was 10 degrees according to plain radiograph. At this writing, the boy is able to maintain a sitting position without any difficulty. CONCLUSIONS: For this child with myelomeningocele, kyphectomy using a surgical threadwire (T-saw) provided a satisfactory result without any major complication. PMID- 12768157 TI - Isolated paramedullary hemangioblastoma originating from the first cervical nerve root: case report. AB - STUDY DESIGN: A retrospective case of an isolated paramedullary hemangioblastoma originating from the first cervical root is reported. OBJECTIVE: To describe an uncommon type of spinal hemangioblastoma and its operative treatment. SUMMARY OF BACKGROUND DATA: Spinal hemangioblastoma, rare finding accounting for approximately 1.5% to 2.5% of all spinal cord tumors, may have an intramedullary, extramedullary, or extradural location. Cervical hemangioblastomas occur in approximately 45% of the cases and are intramedullary in about 83% of the cases. METHODS: A 59-year-old man presented with acute subarachnoid hemorrhage in the basal cisterns. Four-vessel angiography showed a highly vascular small tumor at the dorsolateral side of the cervicomedullary junction fed by a branch of the vertebral artery. The lesion was surgically removed. RESULTS: Total removal of the lesion was achieved after identification of both the arterial feeder and the draining vein with the aid of microvascular Doppler sonography. There were no complications, and the patient did well after surgery. CONCLUSIONS: Although hemangioblastomas occurring in the cervicomedullary area usually may cause progressive neural compression, occasionally they also can present clinically as acute subarachnoid hemorrhage. This situation requires urgent and adequate treatment as in the reported case. PMID- 12768158 TI - Full-thickness thoracic laminar erosion after posterior spinal fusion associated with late-presenting infection. AB - STUDY DESIGN: A case report describes unilateral complete laminar erosion of the caudal thoracic spine and late-presenting infection in a patient 10 years after anteroposterior reconstruction for scoliosis. OBJECTIVE: To present an unusual but significant complication that may occur after implantation of spinal instrumentation. SUMMARY OF BACKGROUND DATA: The reported patient presented with a deep infection and persistent back pain 10 years after successful anteroposterior reconstruction for adult idiopathic scoliosis. Delayed onset infections after implantation of spinal instrumentation are infrequent, yet when present, often require hardware removal. METHODS: The case of a 51-year-old woman who underwent irrigation and debridement for a late-presenting infection and removal of posterior hardware 10 years after her index procedure is presented. Interoperatively, it was noted that full-thickness laminar erosion was present from T4 to T12. RESULTS: The patient was taken to the operating room for wound irrigation, debridement, and hardware removal. It was discovered that a Cotrel Dubousset rod placed on the convexity of the curve had completely eroded through the lamina of T7-T12. Infectious material was found along the entire length of both the convex and concave Cotrel-Dubousset rods. Intraoperative cultures grew Staphylococcus epidermidis and Propionibacterium acnes. Intravenous and oral antibiotics were administered, resulting in resolution of the infection and preoperative pain. CONCLUSIONS: The exact role of late-presenting infection with regard to the laminar erosion and rod migration seen in this case remains to be elucidated. However, the authors believe the primary cause of bony erosion was mechanical in origin. Regardless, most spine surgeons will treat many patients who have had posterior spinal implants and will perform hardware removal on a significant number of these patients during their careers. A full-thickness laminar erosion exposes the spinal cord to traumatic injury during hardware removal and debridement. This case is presented as a cautionary note to help surgeons become cognizant of a potentially devastating complication. PMID- 12768159 TI - Septic arthritis of bilateral lumbar facet joints: report of a case with MRI findings in the early stage. AB - STUDY DESIGN: A case is reported. OBJECTIVE: To report and discuss a case of septic arthritis of bilateral lumbar facet joints with contiguous extension into the paraspinal musculature without infection of the vertebral body or disc space. SUMMARY OF BACKGROUND DATA: Septic arthritis of a lumbar facet joint is a rare cause of low back pain, and only a few cases have been described. If the condition is not associated with epidural or paraspinal abscess formation and self-limiting, the correct diagnosis may be overlooked. To the author's knowledge, septic arthritis of bilateral facet joints has not been previously described in the literature. METHODS: A 72-year-old woman without any systemic illness was hospitalized because of acute severe back pain and high fever. Examination showed local tenderness over the lower lumbar region. Magnetic resonance imaging showed widened facet joints, abscess in the joints, and paraspinal involvement, which aided diagnosis and preoperative planning. RESULTS: The infection did not respond to intravenous antibiotics therapy, so a thorough debridement of the bilateral facet joints was performed. At the latest follow-up assessment, the patient remained afebrile and had a marked reduction of low back pain. CONCLUSIONS: Septic arthritis of the lumbar facet joint is a rare cause of low back pain. It is important to ascertain the diagnosis at the earliest possible stage and to start intravenous antibiotics therapy as soon as possible. Magnetic resonance imaging is a quite sensitive method for identifying infection of the lumbar facet joint. Surgical debridement may be necessary if antibiotic therapy fails. PMID- 12768160 TI - Perioperative drug therapy: a neglected aspect of our practice? PMID- 12768161 TI - Mechanical and infectious complications of central venous catheters. AB - Central venous catheters (CVC) are an important tool in the operation room and intensive care unit. The application of CVC is associated with both mechanical and infectious complications. Knowledge and recognition of risk factors and implementation of strict guidelines will help to reduce the number and severity of complications. Catheter-related factors, patient-related factors, selection of the site of puncture and catheter use and care related factors, all play a contributive role in the risk on complications. PMID- 12768162 TI - Ventilating the patient with severe asthma: nonconventional therapy. AB - Conventional pharmacotherapy of severe asthma and status asthmaticus includes beta2-sympathomimetics, theophylline, corticosteroids and occasionally topical anticholinergics (ipratropium bromide). Since hypoxemia is the most severe phenomenon in status asthmaticus the administration of oxygen is mandatory. However, if the bronchodilating therapy fails and hypoxemia continues, usually respiratory failure develops due to progressive respiratory muscle failure. An increasing PaCO(2) and respiratory acidosis are indications for mechanical ventilatory support to unload the failing respiratory pump. Nowadays, there is increasing consensus that ventilatory support should be administered primarily as non-invasive ventilation (NIV) via a face mask1. However, in a significant number of patients with severe asthma NIV is either contraindicated or insufficient. In this case usually the patient must be endotracheally intubated and mechanically ventilated "invasively". Intubation and ventilation of patients with severe asthma or status asthmaticus is associated with a high incidence of complications compared to patients ventilated for other causes of respiratory failure2,3. Therefore the risks of invasive mechanical ventilation have to be weighted carefully to ongoing conservative therapy and NIV. Cardiopulmonary arrest and severe hypoxemia in spite of O2 supplement and NIV are absolute criteria for intubation and ventilation. Mostly deterioration in mental status and exhaustion are the clinical findings leading to mechanical ventilation. Decision is guided rather by the course of the deterioration (how fast the patient's condition is worsening) than by pathological values alone. An increased PaCO(2) with moderate respiratory acidosis alone is not per se an indication for mechanical ventilation. However, a continuously rising PaCO(2) or the development of a severe metabolic acidosis after 1 hour of NIV is a strong argument for invasive mechanical ventilation. Other criteria are evidence of cardiac failure with fall in pulse volume and dysrhythmias, pneumomediastinum or pneumothorax (which has to be drained before mechanical ventilation!). PMID- 12768163 TI - Acute asthma attack: severity criteria. AB - The authors briefly review the main risk factors for fatal asthma, near-fatal asthma, rapid onset asthma and slow onset asthma. The most recent data in the international literature are also shortly reviewed and discussed. PMID- 12768164 TI - New approaches for the prevention of airway infection in ventilated patients. Lessons learned from laboratory animal studies at the National Institutes of Health. AB - Despite early diagnosis and appropriate antibiotic therapy, ventilator-associated pneumonia (VAP) remains the leading cause of death from hospital-acquired infection in ventilator-dependent patients. Strategies to prevent bacterial colonization of the trachea and lungs are the key to decrease mortality, hospital length of stay, and cost. It is well established that the VAP can result from entry of infected oropharyngeal/gastric secretions into the lower airways. Aspiration may occur during 1) intubation, 2) mechanical ventilation through leakage around the tracheal tube cuff, 3) suctioning of the tracheal tube when bacteria can detach from the biofilm within the tube, or 4) areosolization of bacterial biofilm during mechanical ventilation through the tracheal tube or the ventilator circuit biofilm. From experimental studies in sheep, we drew 3 relevant conclusions: 1) The tracheal tube and neck should be oriented horizontal/below horizontal to prevent aspiration of colonized secretions and subsequent bacterial colonization of the lower respiratory tract. 2) Continuous aspiration of subglottic secretions (CASS) can lower bacterial colonization of the respiratory tract, but at the price of severe tracheal mucosal damage at the level of the suction port. 3) Coating the interior of the tracheal tube with bactericidal agents can prevent bacterial colonization of the tube surface and of the entire respiratory circuit, during 24 hours of mechanical ventilation. PMID- 12768165 TI - Nasal vs oral intubation. AB - Both nasal and oral route for intubation have advantages and disadvantages. Oral intubation is easier to perform, faster and less painful than nasal intubation under direct laryngoscopy, while blind nasal intubation represents a good alternative in conscious patient, without sedation. In trauma patient, oral route should be preferred, with cervical immobilisation. By the contrary, nasal intubation can cause bleeding, retro-pharyngeal and turbinate bones injury, but it seems preferable in preventing laryngeal complications. Moreover nasal intubation seem to increase risk for sinusitis while, there is no clear advantage for any of the two routes, concerning nosocomial pneumonia, bacteriemia and otitis. Nevertheless nasal route increases comfort for the patient and decreases injury and necrosis of tongue and lips; tube fastening is simpler thus reducing accidental extubation. PMID- 12768166 TI - Results of early defibrillation program in Piacenza. AB - Defibrillation as soon as possible is the mainstay of modern emergency system in the treatment of sudden cardiac death. The emergency medical system (EMS) should be integrated with first responders in the community trained to use the semiautomatic external defibrillators (AED). Piacenza Progetto Vita is a European project of early defibrillation through lay first responders integrated within the EMS. After 22 months of the project 1 285 first responders were trained to the use of AED. Survival from sudden cardiac arrest significantly increased (from 3.3% to 10.5%, p<0.01). In particular in the group of patients treated by first responders survival from ventricular fibrillation was 44.1% vs 21.2% of EMS treated group (p < 0.05). A simple training for the use of AED without cardiopulmonary resuscitation training increased survival and created a group of competent AED operator integrated within the EMS. PMID- 12768167 TI - Thrombolytic therapy during or after cardiopulmonary resuscitation. Efficacy and safety of a new therapeutic approach. AB - Acute myocardial infarction (MI) and massive pulmonary embolism (PE) are the underlying causes of cardiac arrest in more than 70% of patients. Thrombolysis is an effective therapy for patients presenting with acute MI or massive PE and has experimentally shown to have beneficial effects on the microcirculatory reperfusion after cardiac arrest, but this treatment has been widely withheld up to now mainly because of the fear of severe bleeding complications. To assess the efficacy and safety of thrombolysis after and during cardiopulmonary resuscitation (CPR), we reviewed the currently available clinical studies on thrombolysis after and during CPR. From these data, there is increasing evidence that thrombolytic therapy during or shortly after CPR can contribute significantly to a restoration of spontaneous circulation in patients suffering from cardiac arrest. Although the use of thrombolytic agents is associated with a higher incidence of bleeding complications, currently available data do not suggest an increase of complications if thrombolysis is combined with CPR. Considering the poor outcome of patients suffering from cardiac arrest and the lack of effective and causal treatment options, the potential risks of thrombolysis after or during CPR probably do not outweigh the benefits of this treatment option. PMID- 12768168 TI - [Some remarks on the management of liver donor]. AB - We evaluated 481 liver donors in order to assess the incidence of positive cultures on samples obtained before harvesting, at harvesting and on preservation fluid; to determine factors related to positive cultures in the donor; to analyse the bacterial and fungal transmission from donor to recipient; to verify the influence of donor culture positivity on graft and patient survival. Cultures were positive in 232 of 481 (48%) donors. Bacteremia was present in 101 of 481 (20%) donors. Intensive care length of stay was significantly longer in culture positive donors. A Gram-negative bacteria transmission from the infected donor to the graft recipient was proven in 1 case. No differences in 1-year survival and retransplantation rates were found between patients receiving livers from culture positive or negative donors. In conclusion, even if rare, donor to host infection transmission is proven. Extended criteria for organ procurement may explain the high number of culture-positive donors we report. Careful microbiological surveillance and treatment can reduce the clinical negative impact on recipient outcome. PMID- 12768169 TI - [Antithrombin: prospects in clinical practice. Sespsi: anticoagulant or anti inflammatory agents?]. AB - Sepsis and septic shock represent a frequent cause of mortality in Intensive Care Units, despite of the progress in antibiotic therapy and in the hemodynamic and respiratory support. The most frequent cause of death is the Multi Organ Dysfunction Syndrome (MODS), which is the clinical manifestation of the irreversibile damage of the microvascular bed. During sepsis and septic shock both activation of coagulation /fibrinolysis and release of mediators of inflammation contribute to the pathogenesis of disseminated intravascular coagulation (DIC); in particular the formation of fibrin in the microvascular bed is the pathological substrate of the clinical development of MODS. The rationale for employing antithrombin (AT) concentrates in the treatment of DIC associated to sepsis is based on the consideration that AT plasma levels are always decreased in patients with sepsis or septic shock; furthermore, the degree of the decrease is directly proportional to the severity of the disease and the prognosis and low AT plasma activities correlate with high mortality. AT has a double function: anticoagulant and anti-inflammatory. The most important mechanism responsible of the anti-inflammatory properties of AT is the binding to the glycosaminoglycans of the endothelial cells and the consequent release of prostacyclin. During sepsis and septic shock, treatment with AT was able, especially in animal models but also in clinical studies, to decrease plasma levels of mediators of inflammation and in some case to preserve organ failure and to reduce mortality. PMID- 12768170 TI - [Antithrombin III in burned children]. AB - We studied haemostatic parameter and Antithrombin III (ATIII) level in 50 consecutive admissions for severe burns between 1990 to1994 to the Centro Grandi Ustionati Ospedale S. Eugenio, Rome. The criteria for inclusion in the study were age <16 years and the presence of major burns (extent of the burns >30 and second and third degree burns). Treatment consisted of initial fluid resuscitation and early excision and grafting of burn wounds and critical care support by a multidisciplinary team. Burn-related variables haemostatic values and the conditions of microcirculation were proposed and analysed with the help of Fisher exact test, chi square test, logistic regression and discriminant analysis. By the discriminant analysis, the coefficients of the standardized functions and the percentage of correctly classified individuals were calculated. The analysis showed total burned surface turned out to be the best predictor of survival. High discrimination efficiency was observed for age, weight, burn type (flame, scald contact chemical electrical). By the logistic regression, the total burned surface confirmed to be the best predictor of survival and between the haemostatic variables ATIII give a significant value (p=0.0244). Moreover, it is noteworthy that ATIII level at the onset of the disease gives a significant correlation with mortality (p=0.0005). The Fisher test showed a significant association between ATIII level and death (p=0.0005). This was confirmed by the chi square test (p=0.00027). Considering the AT deficiency in patients with thermal injuries, we conducted a pilot study to assess AT concentrate infusions for safety and efficacy in thermal injury. The patients received AT concentrate infusions every 8 hours to raise the plasma level to 100 % in the first 72 hours after injury and were compared with control patients with burns. Day4 levels were 105%+/-20% in patients treated with AT patients versus 50%+/-14% in the control patients (P < 0.001). In the group treated with AT the time to microcirculation recovery was shorter (P<0.02). PMID- 12768171 TI - [Emergencies in cardiac surgery: prophylaxis or correction?]. AB - Management of coagulation during cardiac surgery is always challenging for the anesthesiologist, even in elective operations. The strict linkage between coagulation and inflammation is amplified during cardiopulmonary bypass due to the contact of the blood with the foreign surfaces. In emergencies, coagulative derangement could be worse but the cardiocirculatory instability and parenchimal failure often overcome the attention to this problem. PMID- 12768173 TI - [Prevention of hypotension in spinal anaesthesia carried out for caesarean section]. AB - After describing the most commonly applied obstetric indications for caesarean section and the respective percentages reported in countries that are comparable with Italy in terms of health care standards, the clinical reasons and requirements on the basis of which it is considered that spinal anaesthesia is first choice compared to general anaesthesia in obstetrical surgery are outlined. This evidence is confirmed by the spinal anaesthesia/general anaesthesia ratio encountered in the major national and international Obstetric Hospitals. Maternal hypotension remains the most frequent and clinically important complication consequent on spinal anaesthesia in pregnant women at term. The aetiology, frequency and clinical consequences for mother and foetus of severe maternal hypotension and the need to implement effective preventive therapeutic strategies are therefore discussed. The procedures for preventing maternal hypotension are described and for each procedure its validity and standardisation are assessed. It is pointed out that certain procedures have become part of standard practice but their effectiveness has not yet been confirmed while others are not only ineffective but also expose mother and foetus to potential complications. For others again the jury is still out on their real effectiveness. Finally, the techniques that are currently considered to be effective and shared by the majority of authors are described and these must therefore be included in the procedural protocols regarding spinal anaesthesia for caesarean section. PMID- 12768172 TI - OPS techniques. AB - Microcirculatory alterations have been widely described in experimental models of sepsis, however the microcirculation have long been neglected in septic patients as traditional techniques do not allow the visualisation of the microcirculation. The Orthogonal Polarization Spectral (OPS) imaging technique allows the direct visualisation of the microcirculation at the bedside. A selected review of the articles on the microcirculation in patients with sepsis using the OPS imaging technique, is made. Patients severe sepsis present a decrease in capillary density and a decrease in the proportion of the perfused capillaries. The severity of these alterations is more pronounced in non survivors than in survivors, and is related with the development of multiple organ failure. These alterations can be reversed by vasodilators, either topically applied or administered intravenously. Microvascular blood flow alterations are frequently observed in patients with sepsis and can have major pathophysiological implications. PMID- 12768174 TI - Gastric reflux and pulmonary aspiration in anaesthesia. AB - Although aspiration is a relatively rare event during anaesthesia, it represent an important cause of anaesthesia related mortality and also of ventilator associated pneumonia in intensive care unit. The incidence of aspiration is markedly increased after trauma owing to the risk of recent ingestion of food, depression of consciousness and airways reflexes, and gastric stasis induced by raised sympathoadrenal tone. The factors which contribute to the likelihood of aspiration include the urgency of surgery, airways problems, inadequate depth of anaesthetic, use of the lithotomy position, gastrointestinal problems, depressed consciousness, increased severity of illness and obesity. Factors that predispose to aspiration pneumonia are: a gastric content with a pH less than 2.5 and a gastric volume of 0.4 ml kg-1; a reduction in lower oesophageal sphincter tone; a reduction of upper oesophageal sphincter tone and a not coordination between the pharyngeal muscle and the upper oesophageal sphincter tone during swallowing; and a depression of protective airway reflexes. Methods to minimize regurgitation and aspiration involve control of gastric contents (preoperative starvation is the method universal accepted), application of cricoid pressure and control of the airways. PMID- 12768175 TI - Guidelines on anticoagulants and the use of locoregional anesthesia. AB - Growing numbers of patients are treated with hemostasis altering drugs, as thromboembolic diseases are a major cause of mortality in our western society. The insertion of epidural or subarachnoidal needles and/or catheters in such patients carries the inherent risk of the development of a compressing vertebral canal hematoma. This is especially true in patients treated with thrombolytic agents or oral anticoagulants. Extreme caution is also warranted in patients treated with newer compounds as the thienopyridines, glycoprotein IIb/IIIa receptor antagonists, heparinoids, selective factor Xa inhibitors, and direct thrombin inhibitors. The available data do not allow making firm recommendations on the safe use of major neuraxial blocks. In contrast, the isolated use of acetyl-salicylic acid or non-steroidal anti-inflammatory drugs is no longer considered contraindicated, but their combination with of heparin remains controversial. Intraoperative heparinization, perioperative thromboprophylactic use of unfractionated heparin or low molecular weight heparins are possible if: 1) a minimum time interval between the regional anesthetic block and the administration of the previous or next dose of anticoagulant is respected and; 2) the specified dose limitations of the heparin compound used are not exceeded; and 3) indwelling catheters are removed only after the disappearance of any remaining anticoagulant effect. PMID- 12768176 TI - Lowering cardiac risk by preoperative interventions. AB - Postoperative myocardial infarction still represents a serious complication in patients with coronary artery disease, after the first 48 hours following non cardiac surgery (NCS). To reduce the incidence, patient's status should be optimised, including CABG, PTCA and stenting, before the NCS. Indications for CABG, PTCA and stenting suggested by ACC/AHA are in general applicable also for NCS. Patients undergoing low and intermediate risk procedures would not benefit from extensive testing and CABG, while a potential benefit exists for high risk surgery in severe CAD patients. Revascularisation by means of PTCA seems to decrease the risk only if it is performed more than 90 days before the NCS. By the contrary, the combination of PTCA and stenting reduces incidence of coronary re-stenosis, but it increases bleeding, thrombosis and distal embolisation, especially if not supported by an anti-platelet or combined pharmacological schedule; therefore, perioperative stenting is not recommended to increase safety. Moreover, if NCS is urgent, despite a recent stenting, the patient should be included in a high risk group. PMID- 12768177 TI - Anaesthesia for non obstetric surgery in pregnant patients. AB - Female sex constitutes a great part of population and most women are young in childbearing age and expected to be submitted to emergency or urgent obstetric surgery following traumas or diseases which require immediate treatment. Anesthetic considerations for non obstetric surgery during pregnancy include concern for the safety of 2 patients, the mother and fetus, which will be discussed together with the prevention of preterm labor. PMID- 12768179 TI - Cardiovascular and pulmonary effects of epidural anaesthesia. AB - The cardiovascular effects observed with epidural anaesthesia are complex and are predominantly related to the extent of sympathetic denervation, autonomous balance, baseline blood volume and cardiovascular function of the patient. With epidural anaesthesia the pharmacological effect of systemically absorbed local anaesthetic agents and inclusion of adrenaline to the local anaesthetic solution may play a role. Individual cardiovascular response to different levels of sympathetic blockade varies widely, depending on the degree of sympathetic tone prior to the block. Epidural anaesthesia per se has little effect on respiration in patients with pre-existing lung disease. Thoracic epidural anaesthesia appears to reverse the diaphragmatic dysfunction which is a major determinant of the decrease in lung volumes observed after upper abdominal surgery. This article summarizes cardiovascular and pulmonary responses to epidural anaesthesia. PMID- 12768178 TI - [Prevention of cardiovascular accidents during locoregional anesthesia]. AB - Cardiovascular complications can be divided into 2 big categories involving the cardiovascular system in locoregional anaesthesia: those of local anaesthetics with direct effect on sympathetic fibres, which regulate the cardiovascular activity and those who derives from alteration of the normal cardiac function due to the toxic effect of the drugs. While the first are referred to the extension of a central block, the second considers the overdose caused by accidental intravenous injection. This is more frequent in peripheral blocks then in central blocks. The use of catheters in both central and peripheral blocks introduces another factor of risk due to a prolonged administration of local anaesthetics. The prevention of those complications should foresee through an accurate anamnesis the subjective conditions of risk: so we have to choose the best individual technique and dose of anaesthetics; the use of qualitative correct material, the ENS as a support to identify nerve structures and the application of more recent and safe drugs represented by the compound of S(-) enantiomers, Ropivacaine and Levobupivacaine. Which are described to be less cardiotoxic but with the same characteristics as Bupivacaine. Finally don't forget respect the classical rules of security during locoregional anaesthesia. PMID- 12768180 TI - Postanaesthetic shivering. Epidemiology, pathophysiology and approaches to prevention and management. AB - Postanaesthetic shivering is one of the leading causes of discomfort for patients recovering from general anesthesia. During EMG records, the distinguishing factor from shivering in fully awake patients is the existence of clonus similar to that recorded in patients with spinal cord transection. They coexist with the classic waxing and waning signals associated with cutaneous vasoconstriction (thermoregulatory shivering). The causes responsible for their appearance primarily include hypothermia, which sets in due to thermoregulation inhibition by anesthetics. However, we also note the existence of shivering associated with cutaneous vasodilatation (non-thermoregulatory shivering) one of the origins of which is postoperative pain. Apart from the discomfort and aggravated pain, postanaesthetic shivering raises metabolic demand proportionally to the solicited muscle mass and the patient's cardiac capacities. No link has been demonstrated between their occurrence and an increase in cardiac morbidity but it is preferable to avoid postanaesthetic shivering since it is oxygen draining. Prevention mainly entails preventing hypothermia by actively rewarming the patient. Postoperative skin surface rewarming is a way of obtaining the threshold shivering temperature while raising the skin temperature and improving the patient's comfort. However, it is less efficient than certain drugs such as meperidine, nefopam or tramadol, which act by reducing the shivering threshold temperature. PMID- 12768181 TI - Post-operative epidural versus intravenous patient-controlled analgesia. AB - Patient-controlled analgesia techniques have opened a new dimension to individualize patient's need for analgesia, in the treatment of acute post operative pain. These techniques can be used intravenously, in the epidural space, and into peripheral nerve sheets. There is a common consensus that intravenous patient-controlled analgesia should not have a continuous infusion while epidural patient-controlled analgesia (PCEA) should be programmed with a continuous infusion. The drugs used for epidural analgesia are: opioids, local anaesthetics or the combination of both. The combinations seem to provide better pain relief and less side effects. The continuous epidural infusion of opioids has the advantages of fewer fluctuations in cerebrospinal fluid concentrations of drug, but it is necessary to administer a loading bolus, to overcome the fact that it takes several hours to provide adequate analgesia. The advantages of epidural versus intravenous patient-controlled analgesia are represented by better analgesia and a reduced opioids requirement, while the advantages when compared to epidural continuous infusion are: increased efficiency, self adjustment by the patient, higher patient satisfaction, less sedation, and lower opioids dosage. The clinical advantages of PCEA may outweigh the greater cost and invasiveness of this technique. PMID- 12768182 TI - Hypoxia following surgery - an unnecessary cause of morbidity and mortality? PMID- 12768183 TI - Some open questions in pediatric regional anesthesia. AB - The aim of the paper is to review the literature concerning 4 unanswered or debatable questions concerning the practice of regional anesthesia in pediatric patients. The published material concerning the 4 selected topics is reviewed, namely importance of ropivacaine, preoperative coagulation screening tests, hemodynamic stability following neuraxial blocks and prevention/treatment of post dural puncture headache. Of the 4 questions considered in this article, 3 can be reasonably answered in a consensual way. Ropivacaine has limitations for single shot procedures in infants but its advantages for continuous infusions are significant in comparison with those of bupivacaine. Preoperative coagulation screening tests are not necessary, even not useful in children when clinical history is not suggestive of coagulation disorders, with the notable exception of neonates and prematurely born infants less than 45 weeks of post-conceptual age. The long established hemodynamic stability following neuraxial blocks results from well equilibrated compensatory mechanisms which may not be functional in children with preoperative hemodynamic instability or anomalies of the regional blood flow distribution. Finally, even though the post-dural puncture headache is not frequent in children, its management still remains difficult and no definitive recommendation can be currently made in case of inadvertent dural puncture during an attempted epidural anesthesia in children. PMID- 12768184 TI - Upper respiratory tract infections and pediatric anesthesia. AB - Anesthesia for the child with an upper respiratory infection (URI) presents a challenge for the pediatric anesthesiologist. Differences in study design have made interpretation and comparison very difficult. The general lack of evidence based research has led to disparities in the manner in which children which URI have been traditionally managed. Many studies have described associations between URIs and adverse events and we must decide whether to proceed or postpone the procedure and how long to postpone it. More recent research, however, suggests that children with uncomplicated infections can undergo elective procedures without significant increase in adverse anesthetic outcomes. This presentation summarizes the evolving literature about cancellation of surgery for the child with an upper respiratory infection, perioperative outcomes and anesthetic management. PMID- 12768185 TI - [Anesthesia for non-cardiac surgery in children with congenital heart diseases]. AB - The incidence of congenital heart diseases accounts for 8-10 over 1000 liveborn. In Italy about 4000-4500 babies each year are born with congenital heart diseases; 50% of those babies (2000-2200) need cardiac surgery shortly after birth or within the first few months of life. Of the remaining 50%, half undergoes cardiac surgery later on in life and half does not necessitate any surgery; 30% of all cardiac operations consist of palliative procedures and the remaining 70% consist of one-stage corrective procedures. Improvements achieved both in surgical and anesthesiologic techniques, and in cardiopulmonary bypass and myocardial protection, have led to better results in pediatric cardiac surgery, with excellent long term survival rate, even for the more complex variants of congenital heart malformations. Therefore anesthesiologists are now more often required to deal with patients affected by congenital heart defects, for other than cardiac problems. Accurate investigation of patient's clinical history is strongly suggested. Moreover knowledge and familiarity with the modifications of the physiology, occurring in congenital heart disease patients, are mandatory for the choice of the more appropriate anesthesiologic strategy for each patient, in order to optimise the risk-benefits ratio and achieve a less traumatic impact on the cardio-circulatory and respiratory equilibrium. With the aim of achieving better results, interaction between anesthesiologist, cardiologist, pediatrician, surgeon and sometime neonatologist and cardiac surgeon, is strongly recommended in the evaluation of risks, and in decision making of strategies and timing of treatment. PMID- 12768186 TI - Ketamine: a new look to an old drug. AB - Ketamine is an NMDA receptors antagonist, with a potent anaesthetic effect. NMDA receptors are involved in nociceptive modulation, in the wind-up phenomenon, in peripheral receptive fields expansion, in primary and secondary hyperalgesia, in neuronal plasticity. Ketamine effects are well-known: it produces a state of "dissociative anaesthesia", amnesia, and, at the same time, it mantains the respiratory drive effective and supports the sistemic arterial blood pressure. Anaesthesiologists are also familiar with its side-effects, like the increase of salivar and bronchial secretions, the possible increase of intracranial and pulmonary pressures and the dysphoric effect that may produce vivid and sometimes unpleasant dreams. Reviewing scientific data and studies about the use of ketamine in children, many considerations come out: at first they considered the effects of the racemic ketamine, then they evaluated the S-enantiomer. Many surveys studied the effects (analgesia, sedation, side-effects) of different doses or different routes of administration. Other studies were designed to compare ketamine to clonidine or opioids as adjuvants in paediatric regional anaesthesia with local anesthetic drugs, in order to prolong analgesia. In our Children's Hospital, we use ketamine in the operating room, in intensive care unit and for any procedure in hospital wards. The suggested doses are: Epidural or caudal route (as an ajuvant for local anaesthetic agents, in the treatment of postoperative pain): 0.5 mg/kg. Sedative/analgesic effect (for algesic procedures): 1-2 mg/kg i.v. Continuous infusion (intensive care unit): 0.5 mg/kg/h, with a range from 20-30 microg/kg/min to 80 microg/kg/min, depending on the age of the patient. PMID- 12768187 TI - [Anesthesia for neurosurgery in children: techniques and monitoring]. AB - Pediatric neuroanesthesia can be seen as a specific branch of anesthesia half way in between pediatric anesthesia and neuroanesthesia. As a matter of fact, we must keep well in mind the peculiarities of the pediatric patient and the different pharmadynamic and pharmacochinetic properties of the anesthetic drugs, particularly in neonates and infants. Other relevant problems are: 1) high complexity of surgical procedures implying a difficult anesthesiological management; 2) complex blood loss management either if we want to apply a blood sparing technique strategy or if we consider the problems related to diagnosis and treatment of coagulative disorders caused by intraoperative massive blood loss; 3) management of patients with latex allergy for the high incidence, in pediatric neuroanesthesia, of patients belonging to high risk groups; 4) need of repeated radiological examinations implying several anesthesiological procedures. In this article aspects related to the anesthesiological techniques and to the hemodynamic and neurophysiological monitoring of pediatric neurosurgical patients were also discussed. PMID- 12768188 TI - Gene therapy for hepatocellular carcinoma using non-viral vectors composed of bis guanidinium-tren-cholesterol and plasmids encoding the tissue inhibitors of metalloproteinases TIMP-2 and TIMP-3. AB - Metalloproteinases (MMPs) and their natural inhibitors (TIMPs) contribute to the regulation of tumor microenvironment. Their expressions are deregulated in almost all human cancers. We report a novel approach to gene therapy of hepatocellular carcinoma (HCC), using repeated injections of DNA plasmids encoding the tissue inhibitors of metalloproteinases (TIMPs) TIMP-2 or TIMP-3, and a novel competent formulation of gene transfer based on nontoxic cationic cholesterol derivatives. The new gene delivery system was efficient in demonstrating the antitumor efficiency of TIMP-2 or TIMP-3 in inhibiting tumor growth of human HuH7 HCC cells xenografted into nude mice. We show, for the first time, an in vivo effect of TIMP-3 in delaying HCC tumor growth. No treatment-related toxicity was noted. An inhibition of angiogenesis and tumor necrosis accompanied the inhibitory effects of TIMP-2 or TIMP-3 on tumor expansion and invasion. We also report a bystander effect produced by transfected HuH7 tumor cells mixed with untransfected cells in 1:1 ratio in culture that resulted in killing 98% of cells within 96 h. In addition, the soluble forms of TIMP-2 and TIMP-3 expressed by transfected cells exerted a cytotoxic effect on untransfected HuH7 cell cultures. Taken together, these results demonstrate the potential efficacy of repeated treatment of secreted TIMP-2 and TIMP-3 for the design of nonviral gene therapy for hepatocarcinoma. PMID- 12768189 TI - Methioninase gene therapy with selenomethionine induces apoptosis in bcl-2 overproducing lung cancer cells. AB - We have previously shown that the toxic pro-oxidant methylselenol is released from selenomethionine (SeMET) by cancer cells transformed with the adenoviral methionine alpha,gamma-lyase (methioninase, MET) gene cloned from Pseudomonas putida. Methylselenol damaged the mitochondria via oxidative stress, and caused cytochrome c release into the cytosol thereby activating caspase enzymes and thereby apoptosis. However, gene therapy strategies are less effective if tumor cells overexpress the antiapoptotic mitochondrial protein bcl-2. In this study, we investigated whether rAdMET/SeMET was effective against bcl-2-overproducing A549 lung cancer cells. We established two clones of the human lung cancer A549 cell line that show moderate and high expression levels of bcl-2, respectively, compared to the parent cell line, which has very low bcl-2 expression. Staurosporine-induced apoptosis was inhibited in the bcl-2-overproducing clones as well as in the parental cell line. In contrast to staurosporine, apoptosis was induced in the bcl-2-overproducing clones as well as the parental cell line by AdMET/SeMET. Apoptosis in the rAdMET-SeMET-treated cells was determined by fragmentation of nuclei, and release of cytochrome c from mitochondria to the cytosol. A strong bystander effect of AdMET/SeMET was observed on A549 cells as well as the bcl-2-overproducing clones. rAdMET/SeMET prodrug gene therapy is therefore a promising novel strategy effective against bcl-2 overexpression, which has blocked other gene therapy strategies. PMID- 12768190 TI - T-cell-dependent antitumor effects produced by CD40 ligand expressed on mouse lung carcinoma cells are linked with the maturation of dendritic cells and secretion of a variety of cytokines. AB - CD40/CD40 ligand (CD40L) interaction plays an essential role in cell-mediated immune responses. We examined whether expression of CD40L in murine lung carcinoma (A11) cells could produce antitumor effects. The proliferation rate in vitro of A11 cells transfected with the murine CD40L gene (A11/CD40L) was not different from that of parent cells; however, half of the immunocompetent mice inoculated with A11/CD40L cells did not form tumors and the growth of A11/CD40L tumors developed in the rest of mice was significantly retarded compared with that of parent tumors. Protective immunity was also induced in the mice that had rejected A11/CD40L cells. In T-cell-defective nude mice, these antitumor effects were not observed. Bone-marrow-derived dendritic cells (DCs), when cultured with A11/CD40L cells, formed clusters with the tumors and showed upregulated CD86 expression. Expression of the interleukin-23 (IL-23) p19, IL-12p35, IL-18, interferon-gamma (IFN-gamma) and Mig (monokine induced by IFN-gamma) genes was induced in the DCs that were cultured with A11/CD40L but not with A11 cells, and P40, the subunit of both IL-12 and IL-23, was secreted from the cocultured DCs. These data directly showed that the expression of CD40L in tumors facilitated the interaction between DCs and the tumors, enhanced the maturation of DCs, induced secretion of cytokines, and consequently produced T-cell-dependent systemic immunity. PMID- 12768191 TI - Endogenous p21WAF1/CIP1 status predicts the response of human tumor cells to wild type p53 and p21WAF1/CIP1 overexpression. AB - Expression of exogenous wild-type (wt) p53 protein can suppress the growth and/or induce apoptosis in different tumor cells. The effect of exogenous p21(WAF1/CIP1) expression is more controversial: while it can induce apoptosis in some cells, it can protect against p53-mediated apoptosis in others. We used adenoviral vectors to introduce p53 and p21(WAF1/CIP1) genes into human tumor cell lines with different p53 and/or p21(WAF1/CIP1) status. The cell growth inhibition and the induction of apoptosis were measured. Overexpression of wt p53 induced more efficient growth inhibition and apoptosis in SW 620 (mutant p53) and HeLa (inactivated p53 protein) than in MCF-7 (wt p53) and CaCo-2 cell line, which was the most resistant to p53 overexpression despite the p53 mutation. Unlike HeLa and SW 620 cells, the basal p21 protein level was readily detected in CaCo-2 and MCF-7 cells. Overexpression of p21(WAF1/CIP1) gene induced somewhat less pronounced growth inhibition of all cell lines tested, but it also induced apoptosis in HeLa and SW 620 cells. These results suggest that the basal, but not the inducible, levels of p21(WAF1/CIP1) protein in tumor cells could protect from p53-mediated apoptosis. On the other hand, overexpression of p21(WAF1/CIP1) gene itself can induce apoptosis in cells with no basal p21(WAF1/CIP1) protein level. Possible mechanisms of the differential response to these genes are discussed. PMID- 12768192 TI - Requirements for ICAM-1 immunogene therapy of lymphoma. AB - Intercellular cell adhesion molecule-1 (ICAM-1) is a cell-surface glycoprotein capable of eliciting bidirectional signals that activate signalling pathways in leukocytes, endothelial, and smooth muscle cells. Gene transfer of xenogeneic ICAM-1 into EL-4 lymphomas causes complete tumor rejection; however, it is unknown whether the mechanism responsible involves the "foreignness" of the ICAM 1 transgene, bidirectional signalling events, ICAM-1-receptor interaction, or a combination of the latter. To begin to address this question, we constructed four different therapeutic expression vectors encoding full-length ICAM-1, and forms in which the N-terminal ligand-binding domains and cytoplasmic tail had been deleted. Mouse EL-4 tumors (0.5 cm in diameter), which actively suppress the immune response, were significantly inhibited in their growth following injection of expression plasmids encoding either full-length xenogenic (human) ICAM-1, or a functional cytoplasmic domain-deficient form that retains ligand-binding activity. Efficacy of ICAM-1-mediated antitumor immunity was significantly augmented by administration of the antivascular drug 5,6-dimethylxanthenone-4 acetic acid (DMXAA), which suppressed blood supply to the tumor, leading to enhanced leukocyte infiltration, and complete tumor eradication in a gene dosage and CD8(+) T cell and NK cell-dependent fashion. Generation of potent cytotoxic T cell (CTL)-mediated antitumor immunity was reflected by ICAM-1-facilitated apoptosis of tumor cells in situ. In contrast, nonfunctional ICAM-1 lacking the N terminal ligand-binding Ig domain failed to generate antitumor immunity, even in the presence of DMXAA. These studies demonstrate that ICAM-1-stimulated antitumor immunity can overcome tumor-mediated immunosuppression, particularly when employed in combination with an attack on the tumor vasculature. The ligand binding domain of ICAM-1 is essential for generating antitumor immunity, whereas the cytoplasmic domain and bidirectional activation of tumor signalling pathways are not essential. PMID- 12768193 TI - Oncolytic parvovirus H1 induces release of heat-shock protein HSP72 in susceptible human tumor cells but may not affect primary immune cells. AB - Certain autonomous parvoviruses preferentially replicate in and kill in vitro transformed cells and may reduce the incidence of spontaneous and implanted tumors in animals. Hence, these viruses and their derivatives are currently under evaluation as antitumor vectors. However, the mechanisms underlying their tumor suppressing properties are not yet understood. We asked whether the lytic parvovirus H1 may enhance the immunogenicity of infected tumor cells. Out of human melanoma and gastrointestinal tumor cells, we selected the cell line SK29 Mel-1 being very susceptible to H1-induced apoptotic killing. Here, no upregulation of HLA class I and costimulatory molecules could be observed following H1 infection. However, a strong release of the immunogenic signal-the inducible heat-shock protein HSP72, but not constitutive HSP73-was observed after H1 infection. The HSP72 release was higher and of longer duration than a conventional heat-shock treatment. We also explored H1 replication and cytotoxicity in human immune cells, as such cells may constitute targets for H1 virus replication. Long-term cultured lymphocytes, monocytes, immature and mature dendritic cells were not susceptible to H1 virus. Altogether, parvovirus-mediated cell killing may in vivo enhance tumor immunogenicity by HSP72 release and thus contribute to the antitumor effect of parvoviruses. PMID- 12768194 TI - Regression of AK7 malignant mesothelioma established in immunocompetent mice following intratumoral gene transfer of interferon gamma. AB - Malignant mesothelioma (MM) is a lethal tumor linked with a prior exposure to asbestos in which limited progress has been made so far using conventional therapies. MM is an example of a "nonimmunogenic" tumor characterized by a fibrous stroma and an absence of infiltrating T lymphocytes. High levels of transforming growth factor-beta (TGF-beta) produced by mesothelioma cells have been related to the immune tolerance towards the tumor. In order to evaluate the effect of local delivery of cytokines such as interferon gamma (IFN-gamma) by gene transfer, we characterized and used a murine model, AK7, which appeared very similar to human mesothelioma. AK7 cells expressed low levels of major histocompatibility class I and class II antigens and secreted high levels of latent TGF-beta. The TGF-beta pathway in AK7 cells is operative but inefficient because endogenous TGF-beta is predominantly inactive. Treatment of pre established AK7 tumors by direct intratumoral injection of an adenovirus vector expressing murine IFN-gamma, Ad.mIFN-gamma, led to significant tumor regression. Peripheral tumor infiltration by CD4+ and CD8+ T lymphocytes in the treated tumors appeared to be because of the induction of an immune response. Tumor relapse was observed, which could be due to local TGF-beta secretion by remaining tumor cells. PMID- 12768195 TI - Intramuscular gene transfer of soluble B7.1/IgG(1) fusion cDNA induces potent antitumor immunity as an adjuvant for DNA vaccination. AB - Soluble B7.1/IgG Fc fusion protein, which has costimulatory effects, is an effective molecular adjuvant in tumor immune therapy. Here, we describe a nonviral intramuscular (i.m.) gene transfer method to deliver this therapeutic protein. Gene transfer was greatly enhanced by electroporation and highly efficient production of this protein was achieved. Serum levels reached up to 1 microg/ml with considerable length of expression and without apparent systemic adverse effects. Lymphocytes from mice coinjected with soluble B7.1/IgG(1) and carcinoembryonic antigen (CEA)-encoding plasmids showed significantly elevated CEA-stimulated proliferation, cytokine production, and cytotoxic T-lymphocyte (CTL) activity. These mice gained significant protection against a CEA-positive transplanted tumor, in terms of reduced tumor incidence and growth. The effects were superior when soluble B7.1/IgG(1) was expressed as compared to membrane bound wild-type B7.1. Notably, expression of soluble B7.1/IgG(1) alone did not induce any protection against tumor, confirming its primary role as a costimulatory molecule rather than a direct antitumor agent. The plasmid encoding B7.1/IgG(1) did not have to be injected at the same site as the antigen-encoding plasmid to exert its adjuvant effect, indicating that circulating protein was sufficient. Muscle histopathology revealed minimal damage to DNA-injected muscles. Importantly, we show that, after gene transfer, muscle tissue can produce this protein in large quantity to exert its immune costimulatory effect for cancer therapy and it would be otherwise difficult and expensive to maintain this high a level of recombinant protein. PMID- 12768196 TI - Ion transport: tracing the path. PMID- 12768199 TI - Genetic code expansion. PMID- 12768200 TI - Visualizing an mRNA destruction line. PMID- 12768201 TI - SNARE function revisited. PMID- 12768202 TI - More than one way to skin a cat: translational autoregulation by ribosomal protein S15. PMID- 12768203 TI - A charged view of voltage-gated ion channels. PMID- 12768205 TI - Insights into IgA-mediated immune responses from the crystal structures of human FcalphaRI and its complex with IgA1-Fc. AB - Immunoglobulin-alpha (IgA)-bound antigens induce immune effector responses by activating the IgA-specific receptor FcalphaRI (CD89) on immune cells. Here we present crystal structures of human FcalphaRI alone and in a complex with the Fc region of IgA1 (Fcalpha). FcalphaRI has two immunoglobulin-like domains that are oriented at approximately right angles to each other. Fcalpha resembles the Fcs of immunoglobulins IgG and IgE, but has differently located interchain disulphide bonds and external rather than interdomain N-linked carbohydrates. Unlike 1:1 FcgammaRIII:IgG and Fc epsilon RI:IgE complexes, two FcalphaRI molecules bind each Fcalpha dimer, one at each Calpha2-Calpha3 junction. The FcalphaRI-binding site on IgA1 overlaps the reported polymeric immunoglobulin receptor (pIgR) binding site, which might explain why secretory IgA cannot initiate phagocytosis or bind to FcalphaRI-expressing cells in the absence of an integrin co-receptor. PMID- 12768206 TI - POT1 as a terminal transducer of TRF1 telomere length control. AB - Human telomere maintenance is essential for the protection of chromosome ends, and changes in telomere length have been implicated in ageing and cancer. Human telomere length is regulated by the TTAGGG-repeat-binding protein TRF1 and its interacting partners tankyrase 1, TIN2 and PINX1 (refs 5-9). As the TRF1 complex binds to the duplex DNA of the telomere, it is unclear how it can affect telomerase, which acts on the single-stranded 3' telomeric overhang. Here we show that the TRF1 complex interacts with a single-stranded telomeric DNA-binding protein--protection of telomeres 1 (POT1)--and that human POT1 controls telomerase-mediated telomere elongation. The presence of POT1 on telomeres was diminished when the amount of single-stranded DNA was reduced. Furthermore, POT1 binding was regulated by the TRF1 complex in response to telomere length. A mutant form of POT1 lacking the DNA-binding domain abrogated TRF1-mediated control of telomere length, and induced rapid and extensive telomere elongation. We propose that the interaction between the TRF1 complex and POT1 affects the loading of POT1 on the single-stranded telomeric DNA, thus transmitting information about telomere length to the telomere terminus, where telomerase is regulated. PMID- 12768207 TI - Functional proteomic identification of DNA replication proteins by induced proteolysis in vivo. AB - Evolutionarily diverse eukaryotic cells share many conserved proteins of unknown function. Some are essential for cell viability, emphasising their importance for fundamental processes of cell biology but complicating their analysis. We have developed an approach to the large-scale characterization of such proteins, based on conditional and rapid degradation of the target protein in vivo, so that the immediate consequences of bulk protein depletion can be examined. Budding yeast strains have been constructed in which essential proteins of unknown function have been fused to a 'heat-inducible-degron' cassette that targets the protein for proteolysis at 37 degrees C (ref. 4). By screening the collection for defects in cell-cycle progression, here we identify three DNA replication factors that interact with each other and that have uncharacterized homologues in human cells. We have used the degron strains to show that these proteins are required for the establishment and normal progression of DNA replication forks. The degron collection could also be used to identify other, essential, proteins with roles in many other processes of eukaryotic cell biology. PMID- 12768208 TI - [Helicobacter pylori in Peru]. PMID- 12768209 TI - [Hepatitis C: national studies]. PMID- 12768210 TI - [Percutaneous drainage of hepatic pyogenic abscess: management efficacy]. AB - OBJECTIVE: To show the success of percutaneous drainage combined with an antibiotic therapy in the management of hepatic pyogenic abscess. EQUIPMENT AND METHODS: Health histories of 24 patients diagnosed with hepatic pyogenic abscess were evaluated in the Unit of Vascular and Intervention Radiology (URVI) of the Eduardo Rebagliatti Martins Hospital and were checked, during the time period beginning in January 2001 and ending in June 2002. 23 patients underwent percutaneous drainage, guided by echography. RESULTS: A total of 36 abscesses were found, with an average diameter of 6,78 cm (3-18cm); the most common location was on the right (78%). In 37,5% of the patients, the cause of the abscess could not be determined; in 33,3%, the cause was determined after surgical intervention, primarily cholecystectomy (12,5%). Pseudomona (12,5%) was the species most found in cultivation. Only 28 abscesses were drained percutaneously. On average, drainage lasted 15,8 days, and there was an average of 3,6 controls per patient. There was 89,30% overall success for the procedure with three documented errors. CONCLUSION: Percutaneous drainage in conjunction with proper antibiotic coverage is efficient in the management of hepatic pyogenic collections, and its use must be generalized. PMID- 12768211 TI - [Hepatitis C virus infection in blood bank donors at the Hospital Nacional Cayetano Heredia (1998 - 2002)]. AB - Hepatitis C virus infection (HCV) is the main cause for chronic hepatic disease around the world. Nowadays, there are highly sensitive and specialized tests such as third generation ELISA, used to detect anti-HCV antibodies and to prevent its transmission. Since 1998, this test has been applied to blood donors in our hospital. The objectives of this study were to determine the predominance and some epidemiological characteristics of the HCV infection in blood donors at the Cayetano Heredia National Hospital (C.H.N.H.). This is a descriptive and retrospective study; the epidemiological facts and general sieve results were obtained from the Blood Bank register from 1998 to June 2002. The third generation ELISA test was used. During the time period evaluated, 15,009 blood donors were examined and 122 positive cases were found (0.813% prevalence). The yearly prevalence in 1998 was 14 cases in 2960 donors (0,47%); in 1999, 25 cases in 3136 donors (0,79%); in 2000, 46 cases in 3311 donors (1,38%); in 2001, 21 cases in 3730 donors (0,56%); and in 2002, 16 cases in 1872 donors (0,85%). Likewise, 97 cases (79,5%) were males and 25 cases (20,5%) were females. Seroprevalence predominated in the age group from 21 to 30 (36,06%), followed by the age group from 31 to 40 (27,86%). 47 cases (38,5%) came from the Districts of Lima, Comas and San Martin de Porres. With respect to risky conducts, 79 cases (64,5%) reported having sexual relations without protection, 15 cases (12,29%) reported sexual relations with prostitutes, and only 4 cases (3,27%) reported having received blood transfusions before. In conclusion, the prevalence of the HCV infection in blood donors at the C.H.N.H. is 0,813% from 1998 to June 2002, and sexual relations without protection is the most frequent characteristic. PMID- 12768212 TI - [Human blastocystosis: prospective study symptomatology and associated epidemiological factors]. AB - An attempt has been made to contribute to the understanding of the symptoms and factors associated with the Blastocystis Hominis infection, as seen in persons seeking outside consultation from the Dermatological and Transmissible Diseases Department (DTDD) at the C.H.N.H. This is a case-control study carried out in people between the ages of 5 and 80 in a period from January to March 1999. The cases tested positive in parasitological tests for Blastocystis Hominis and were absent of other enteropathogens. The controls tested negative in parasitological tests for Blastocystis Hominis and were absent of other enteropatoghens. A clinical chart was used to register details of symptomatology and factors associated with the Blastocystis Hominis infection. 74 cases and 70 controls were studied, matched by sex and age. A statistical correlation was obtained (p<0.05) among symptomatic persons and presence of Blastocystis Hominis (91,9%). The symptomatology associated with the Blastocystis Hominis infection by order of statistical significance (p<0.05) was: Abdominal pain (OR=3) 1.47nicotine (22%) approximately morphine (19%) approximately caffeine (19%) approximately methylphenidate (22%) >tomoxetine (9%). Tomoxetine (noradrenergic preferring) was the only compound that did not produce a significant improvement in accuracy. CONCLUSIONS: These results provide pharmacological support for the concept that attentional mechanisms may play an important role in the "environmental" associative aspects of drug seeking behavior, and as such they may provide the basis for treatment strategies aimed at preventing relapse in detoxified addicts. PMID- 12768268 TI - Locomotor and discriminative-stimulus effects of cocaine in dopamine D5 receptor knockout mice. AB - RATIONALE: Dopamine D1-like antagonists block several effects of cocaine, including its locomotor-stimulant and discriminative-stimulus effects. Because these compounds generally lack selectivity among the dopamine D1 and D5 receptors, the specific roles of the subtypes have not been determined. OBJECTIVES: Dopamine D5 receptor knockout (DA D5R KO), heterozygous (HET) and wild-type (WT) mice were used to study the role of D5 dopamine receptors in the effects of cocaine. In addition, effects of the D1-like antagonist, SCH 39166 were also studied to further clarify the roles of D1 and D5 dopamine receptors in the discriminative-stimulus effects of cocaine. METHODS: DA D5R KO, HET and WT mice were treated with cocaine (3-30 mg/kg) or vehicle and their horizontal locomotor activity was assessed. The mice were also trained to discriminate IP injections of saline from cocaine (10 mg/kg) using a two-lever food-reinforcement (FR10) procedure. Doses of cocaine (1.0-10 mg/kg) were administered 5 min before 15-min test-sessions. RESULTS: Cocaine dose-dependently stimulated activity in each genotype, with the highest level of activity induced in the DA D5R WT mice. Both DA D5R KO and HET mice showed reduced levels of horizontal activity compared to WT mice. All three genotypes acquired the discrimination of 10 mg/kg cocaine; doses of 1.0-10.0 mg/kg produced dose-related increases in the number of cocaine appropriate responses. SCH 39166, at inactive to fully active doses (0.01-0.1 mg/kg) produced predominately saline-appropriate responding. SCH 39166 produced a dose-dependent rightward shift in the cocaine dose-effect curve in all genotypes, with similar apparent affinities. CONCLUSIONS: The present data suggest an involvement of DA D5R in the locomotor stimulant effects of cocaine. In addition, the data indicate that there is little involvement of the DA D5R in the discriminative-stimulus effects of cocaine. In addition, the antagonism data suggest a role of the D1 receptor in the behavioral effects of cocaine. PMID- 12768269 TI - Long-lasting effects of chronic stress on DOI-induced hyperthermia in male rats. AB - RATIONALE: Exposure to chronic stress can affect the serotoninergic (5-HT) system and behavioral measures associated with 5-HT. Repeated stress increases 5-HT receptor subtype 2 (5-HT2) mediated behaviors in rodents, such as wet dog shakes and head twitch. OBJECTIVES: The current study investigated whether exposure to chronic unpredictable stress would augment 5-HT(2A/C) receptor-mediated hyperthermia. Furthermore, the persistence of these hyperthermic effects was investigated by testing rats up to 60 days after the stress procedure terminated. METHODS: For 2 or 10 days, rats were either not stressed (controls) or exposed to chronic unpredictable stress, i.e. two stressors per day of the following: cage rotation, cold exposure, swim, restraint, light cycle manipulations, single housing, and food and water deprivation. After the termination of stress (day 3 or 11), the 5-HT(2A/C) receptor agonist DOI (1.5 mg/kg) or saline, was injected and the rectal temperature of the rats was monitored. In a separate experiment, the 5-HT2 receptor antagonist, LY-53,587, was injected 30 min prior to the injection of DOI or saline. Finally, DOI was injected into rats 8, 30 or 60 days after the 10-day stress procedure ended. RESULTS: Rats exposed to 10 days, but not 2 days, of unpredictable stress exhibited higher rectal temperatures following DOI than non-stressed rats. The DOI-induced hyperthermia was attenuated by LY-53,587. The augmentation of DOI-induced hyperthermia in stressed rats persisted when examined 8, 30 and 60 days following the stress procedure. CONCLUSIONS: The enhancement of 5-HT receptor function by chronic stress persists even after the environmental stressor is removed. This lasting increase in 5-HT receptor function may have implications for clinical disorders associated with stress, such as depression or post-traumatic stress disorder. PMID- 12768270 TI - Curricular approaches to research ethics training for psychiatric investigators. AB - RATIONALE: Training in research ethics is crucial for psychiatric investigators. Addressing ethical dimensions of human subjects research requires knowledge about the rules and norms governing research; sensitivity to ethical implications of actions; and skills in ethics problem solving. Investigators in training who are physicians have the additional challenge of developing identities as investigators that sometimes conflict with their identities as physicians. OBJECTIVES: To propose a curriculum for psychiatric research ethics training. METHODS: Review of literature on ethics education and presentation of a curricular approach to research ethics training for psychiatric investigators. RESULTS: Research ethics can be learned and should be taught. Involvement of active investigators in teaching research ethics is important. While core topics of psychiatric research ethics training have not yet been identified, there are available models from which to draw. Research ethics should be introduced early and integrated throughout the research training period. Lack of resources and institutional support can be obstacles to development of comprehensive research ethics curricula. Small-group, case-based discussion is best for teaching ethics problem-solving skills. Examples of teaching module ideas and a fully developed sample teaching module are presented. CONCLUSIONS: There is opportunity for creative models for teaching psychiatric research ethics. Work is needed to identify core topics, target pedagogical strategies to trainees at different levels, and develop evaluation methods. PMID- 12768271 TI - Problem areas in the understanding of informed consent for research: study of middle-aged and older patients with psychotic disorders. AB - RATIONALE: Optimizing the abilities of individuals with psychiatric disorders to provide meaningful informed consent for research has become a heightened concern for psychiatric researchers. OBJECTIVES: We examined a post-consent test of comprehension given to older patients with psychotic disorders to identify problem areas in the understanding of informed consent for research. METHODS: One hundred and two middle-aged and older outpatients with schizophrenia or related psychotic disorders and twenty normal comparison subjects were administered a 20 item questionnaire to assess their comprehension of consent for a low-risk research protocol after receiving either a routine (paper-based) or an enhanced (computerized, structured slide show incorporating greater review) consent procedure. Data on individual questions were analyzed. RESULTS: Patients had more difficulty than normal comparison subjects on open-ended questions, including those asking about study procedures, time involved, and potential risks and benefits. Among patients, the enhanced procedure was associated with better performance on questions about potential risks and time required than the routine procedure. CONCLUSIONS: Problem areas in the understanding of informed consent, such as study procedures and potential risks and benefits, should be the focus of attempts to improve the consent process for patient participants with severe mental illness. Research also should be done to clarify how best to assess understanding of consent, since the wording of questions likely affects the responses. PMID- 12768272 TI - Involvement of noradrenergic system within the central nucleus of the amygdala in naloxone-precipitated morphine withdrawal-induced conditioned place aversion in rats. AB - RATIONALE: We previously reported that the central nucleus of the amygdala (CeA) plays a crucial role in the negative affective, rather than somatic, component of morphine withdrawal. However, numerous studies have reported that the central ascending noradrenergic system is implicated in morphine withdrawal syndrome, although the roles of the noradrenergic system within the CeA in the negative affective component remains less clear. OBJECTIVES: The possible role of the noradrenergic system within the CeA in the negative affective component of morphine withdrawal was investigated. METHODS: The extracellular noradrenaline level within the CeA during naloxone-precipitated morphine withdrawal was measured using an in vivo microdialysis experiment on unanesthetized and freely moving rats. The effects of microinjection of beta-adrenoceptor antagonists into the bilateral CeA on the naloxone-precipitated morphine withdrawal-induced conditioned place aversion (CPA) and somatic signs were examined. RESULTS: The extracellular noradrenaline level within the CeA was transiently elevated during morphine withdrawal. Intra-CeA injections of beta-adrenoceptor antagonists propranolol (30 nmol per side) and timolol (10 nmol per side) significantly attenuated the morphine withdrawal-induced CPA. Similarly, beta(1)-antagonist atenolol (30 nmol per side) or beta(2)-antagonist butoxamine (30 nmol per side) significantly attenuated the CPA. In contrast, they did not affect morphine withdrawal-induced somatic signs, except for propranolol. CONCLUSION: These results suggest that the activation of the noradrenergic system within the CeA contributes to naloxone-precipitated morphine withdrawal-induced CPA, rather than somatic signs, through beta(1)- and beta(2)-adrenoceptors. PMID- 12768273 TI - A collaborative model for research on decisional capacity and informed consent in older patients with schizophrenia: bioethics unit of a geriatric psychiatry intervention research center. AB - RATIONALE: The numbers of older persons with psychiatric disorders are expected to rise rapidly in coming decades, yet most studies of the safety and efficacy of treatments for such disorders have focused on younger adults. A substantial expansion in research involving older patients is needed to meet the treatment needs of this fast growing group. A critical issue in intervention research is ensuring a patient's decision-making capacity. Considerable heterogeneity exists in this regard even within diagnostic groups. Cognitive changes as well as increased complexity of medication regimens in elderly patients may make it particularly difficult for some older persons to fully understand, appreciate, and/or reason about the risks and benefits of participating in any particular study. OBJECTIVES: Empirical research into assessing and possibly improving decisional capacity is warranted in older people with severe mental illness. Such research may be accomplished through collaborations among specialists from various related disciplines and, importantly, with active involvement of community partners. METHODS: We present one model of this type of collaboration, the Bioethics Unit of an Intervention Research Center, comprising a multi disciplinary team along with a Community Advisory Board. RESULTS: Preliminary studies in our Center suggest that older individuals with psychotic disorders vary considerably in their decisional capacity, and many subjects appear to be fully capable for consenting to research projects. Furthermore, the patients' level of understanding of the consent material can be improved significantly through repetition and clarification of key elements in the consent form. CONCLUSIONS: The decisional capacity for a given research protocol is not necessarily an unmodifiable trait, but can be enhanced with improvements in consenting procedures, even in older persons with psychotic disorders. PMID- 12768275 TI - cAMP-dependent protein kinase and reward-related learning: intra-accumbens Rp cAMPS blocks amphetamine-produced place conditioning in rats. AB - RATIONALE: Dopamine may produce reward-related learning by activating D(1)-like receptors in the nucleus accumbens (NAc) and stimulating the formation of cyclic adenosine monophosphate (cAMP) and the activation of cAMP-dependent protein kinase (PKA). OBJECTIVES: This hypothesis was tested using the conditioned place preference (CPP) based on NAc injections of amphetamine (amph) and evaluating the effects of PKA inhibition with Rp-cAMPS. METHODS: The CPP procedure consisted of three phases: pre-exposure (three 15-min sessions in a chamber consisting of two distinct compartments connected by a tunnel), conditioning (four 30-min placements into one compartment with the tunnel blocked following drug injection into the NAc alternating with four similar placements into the other side following NAc injection of saline), and test (one 15-min session with the tunnel open). A CPP was defined as an increase in time spent on the drug-paired side from mean pre-exposure to test. RESULTS: Dose-response experiments showed that 15.0 or 20.0 but not 5.0 or 10.0 micro g/0.5 micro l per side of amph produced a CPP. The amph (20.0 micro g) CPP was blocked by Rp-cAMPS co-injections of 25.0 and 250 but not 2.5 ng/0.5 micro l per side. Rp-cAMPS or the PKA activator Sp cAMPS (50.0, 250, 500, 600 ng/0.5 micro l per side) alone had no effect on side preference. Co-injection of 10.0 micro g amph+Sp-cAMPS (25.0, 50.0, 250, 500 ng) did not result in a CPP but co-injection of 20.0 micro g amph+Sp-cAMPS (250 ng) led to a loss of the CPP normally seen with that dose of amph. Doses of Rp-cAMPS that blocked CPP did not block the locomotor stimulatory effect of amph during conditioning sessions. CONCLUSIONS: Results supported the hypothesis that PKA activation in NAc is necessary for reward-related learning. PMID- 12768274 TI - On the role of noradrenaline in psychostimulant-induced psychomotor activity and sensitization. AB - RATIONALE: Psychostimulant drugs exert their behavioral effects primarily through enhancement of monoaminergic neurotransmission. Augmented dopamine activity is thought to play a critical role in the psychomotor stimulant effects of amphetamine and cocaine, as well as in the development of long-term behavioral sensitization evoked by repeated exposure to amphetamine. However, despite the fact that brain dopamine and noradrenaline systems are closely interconnected, the extent to which noradrenergic transmission contributes to these behavioral effects of psychostimulants is a relatively unexplored issue. OBJECTIVES: By inhibiting noradrenergic neurotransmission with the alpha2-adrenoceptor agonist clonidine, the alpha1-antagonist prazosin and the beta-antagonist propranolol, we investigated the involvement of noradrenaline neurotransmission in the psychomotor stimulant and long-term sensitizing effects of d-amphetamine and cocaine in rats. METHODS: Clonidine (0.003-0.1 mg/kg), prazosin (0.1-3.0 mg/kg) and propranolol (1.0-3.0 mg/kg) were administered prior to d-amphetamine (1.0 mg/kg), cocaine (15 mg/kg) or apomorphine (1.0 mg/kg) and psychomotor activity was measured. In separate studies, clonidine (0.03 mg/kg), prazosin (1.0 mg/kg) or propranolol (3.0 mg/kg) were co-administered with d-amphetamine (2.5 mg/kg) or cocaine (30 mg/kg) for 5 days, and locomotor sensitization was assessed 3 weeks post-treatment. RESULTS: The psychomotor stimulant effect of d-amphetamine, but not that of cocaine or apomorphine, was dose-dependently inhibited by clonidine and prazosin, and enhanced by propranolol. Clonidine, prazosin, and propranolol did not influence the induction of sensitization by amphetamine or cocaine. CONCLUSIONS: Enhancement of synaptic noradrenaline concentrations contributes to the psychomotor stimulant effect of d-amphetamine, but not cocaine or apomorphine. In addition, noradrenergic neurotransmission is not critically involved in the induction of psychostimulant sensitization. PMID- 12768277 TI - Treatment improves serotonin transporter binding and reduces binge eating. AB - RATIONALE: Serotonin (5-HT) is involved in the control of eating behaviour by inhibiting food intake. Obese women with binge-eating disorder (OB-BED) were recently found to have reduced 5-HT transporter binding. OBJECTIVES: The aim of this study was to investigate the effect of a successful treatment on 5-HT transporters in OB-BED. METHODS: The 5-HT transporter binding of seven OB-BED was measured by single-photon emission computed tomography (SPECT), by using iodine 123-labelled nor-beta-CIT as a tracer, before treatment and after successful treatment, when the OB-BED were asymptomatic. Treatment consisted of group psychotherapy and fluoxetine medication. The control subjects, six obese women without eating disorders, were also studied twice by using SPECT. RESULTS: The 5 HT transporter binding of the symptomatically recovered OB-BED increased significantly (24+/-22%) after treatment, whereas in the control group, binding remained unchanged. CONCLUSIONS: The results tentatively suggest that 5-HT transporter binding in OB-BED is an adaptive mechanism, which can be affected by treatment. Furthermore, there seems to be a link between improved 5-HT transporter binding and reduced binge eating. PMID- 12768276 TI - The 5-HT1A receptor agonist MKC-242 reverses isolation rearing-induced deficits of prepulse inhibition in mice. AB - RATIONALE: Prepulse inhibition (PPI) of startle provides an operational measure of sensorimotor gating in which a weak stimulus presented prior to a startling stimulus reduces the startle response. PPI deficits observed in schizophrenia patients can be modeled in rats by individual housing from weaning until adulthood. The deficits in PPI produced by isolation rearing can be reversed by antipsychotics. We previously found that (S)-5-[3-[(1,4-benzodioxan-2 ylmethyl)amino]propoxy]-1,3-benzodioxole HCl (MKC-242), a highly potent 5-HT(1A) receptor agonist, reduced aggressive behavior selectively in isolation-reared mice. OBJECTIVE: This study examines whether isolation rearing of mice produces PPI deficits and whether PPI deficits are attenuated by 5-HT(1A) receptor activation. METHODS: Male ddY mice, 4 weeks old, were housed for more than 6 weeks singly or in groups of five or six. The PPI of the acoustic startle response was measured using SR-LAB systems. RESULTS: The PPI was less in isolation-reared mice than in group-reared mice. Oral administration of MKC-242 at 0.1-0.3 mg/kg reversed PPI deficits in isolation-reared mice, although it did not affect PPI in group-reared mice. MKC-242 did not affect MK-801-induced and apomorphine-induced PPI deficits in group-reared mice. The reversal by MKC-242 of isolation-induced PPI deficits was antagonized by the 5-HT(1A) receptor antagonist WAY100635 at low doses. CONCLUSION: These results suggest that isolation rearing produces deficits in sensorimotor gating in mice that are reversible by activation of 5-HT(1A) receptors, probably somatodendritic 5-HT(1A) autoreceptors. PMID- 12768278 TI - Bone mineral density may be related to atherosclerosis in hemodialysis patients. AB - Biological interactions between the bone and the blood vessels are gradually being clarified. To investigate the relationship between bone mineral density and atherosclerosis in hemodialysis patients, we examined the bone mineral density and the intima-media thickness of the carotid artery in 83 dialysis patients with non-diabetic nephropathy (44 men and 39 women) aged from 23 to 83 years. The duration of hemodialysis ranged from 2 to 344 months. The bone mineral density of the radius was measured by dual-energy X-ray adsorptiometry, and the ratio of this value to the standard value for the same age and gender was calculated ( Z score). As an index of atherosclerosis, the intima-media thickness of the carotid artery was measured by high resolution B-mode ultrasonography. Then the relationship between the Z-score and various factors was examined using Spearman's rank correlation analysis and multiple regression analysis. The Z score showed a negative correlation with the duration of hemodialysis, the carotid intima-media thickness, and the levels of alkaline phosphatase, intact parathyroid hormone, and low-density lipoprotein cholesterol by Spearman's rank correlation analysis. In addition, the Z-score showed a positive correlation with the lipoprotein (a) level and a negative correlation with the duration of hemodialysis, intima-media thickness, intact parathyroid hormone, and low-density lipoprotein cholesterol by multiple regression analysis. These findings suggest that the decrease of bone mineral density in hemodialysis patients is correlated with secondary hyperparathyroidism and hyperlipidemia, which are factors known to promote atherosclerosis, and thus bone density changes might be related to the progression of atherosclerosis, or vice versa. PMID- 12768279 TI - Basic fibroblast growth factor forms new trabeculae that physically connect with pre-existing trabeculae, and this new bone is maintained with an anti-resorptive agent and enhanced with an anabolic agent in an osteopenic rat model. AB - Osteoporosis is a disease of excess bone fragility that results from both the loss of bone mass and trabecular bone microarchitecture, thereby creating a very fragile skeleton. The purpose of this study was to determine whether treatment of ovariectomized (OVX) osteopenic rats with basic fibroblast growth factor (bFGF) would stimulate the production of new trabeculae, and whether the newly formed trabeculae would make physical connections with the pre-existing trabeculae after prolonged estrogen deficiency. Six-month-old Sprague Dawley rats were OVXed or sham-operated and were left untreated until day 60 post-OVX. A high resolution microscopic scan (XTM) of the right proximal tibia was performed on groups 1 and 2 on day 1 post-OVX, and was repeated in all animals on day 60 post-OVX. At day 60 groups 1 and 2 were treated with vehicle and groups 3 to 6 were injected with bFGF 200 microg/kg/d intravenously for 15 days. At day 82, all animals obtained another in vivo XTM scan of the right tibia; then group 4 were treated with 17B estradiol 10 microg/kg/3x a week, group 5 were treated with hPTH (1-34) at 80 microg/kg/d for 35 days, group 6 were sacrificed, and groups 1 and 2 were treated with vehicle injections for 35 days. At day 110, all remaining animals were sacrificed, and repeat ex vivo XTM scans of the right proximal tibia were performed. Trabecular bone structural variables-including trabecular bone volume, connectivity, number, and thickness-were obtained from all XTM scans. Biochemical markers of bone turnover were also obtained 24 hours before each XTM scan (osteocalcin and deoxypyridinoline), and analyzed by ELISA. Animals OVXed and treated with vehicle had decreased trabecular bone volume, connectivity and number compared to sham-operated animals at both day 60 and day 110. Animals treated with bFGF from day 60-75 post-OVX had evidence of new trabeculae that physically connected with pre-existing trabeculae and also of increased trabecular bone volume seven days after the injections were discontinued. Biochemical markers of bone formation had a small and insignificant increase over baseline levels during the bFGF injections. Bone resorption markers were significantly reduced during the injection period, but returned to baseline levels after the injections were stopped. In addition, we also demonstrated that these newly formed trabecular connections could be maintained or added to with either estrogen or hPTH (1-34) treatments. Thirty-five days after ending the bFGF treatment, trabecular bone volume and connectivity was 25-80% higher in the estrogen and hPTH (1-34) treated animals compared to the untreated animals ( p<0.01). These results support continued development of bFGF as a potential treatment for severely osteoporotic individuals. PMID- 12768280 TI - Discrepancies between hip BMD and fracture rate in Turkey: a familiar problem. PMID- 12768281 TI - Occupational exposure to n-hexane in Italy--analysis of a registry of biological monitoring. AB - OBJECTIVE: This study assesses 2,5-hexanedione (2,5-HD) in the urine of subjects exposed to n-hexane solvent between 1991 and 1998, from details obtained from the Registry of Biological Monitoring (BM) at the Florence Local Health Unit, and its development over time. METHODS: The Registry contains 15,925 samples from 6,650 subjects occupationally exposed to n-hexane, especially in leather (9,099 samples; 3,607 subjects) and shoe (3,865 samples; 1,938 subjects) production. RESULTS: Over the time span studied there was a total reduction of 31.9% in urinary 2,5-HD level. The yearly decrease over the entire period was 5.4%. Dividing the 8 years into three periods: before the introduction of the new legislation for health protection in the workplace (1991-1993), during its transition (1994-1996) and after its complete enforcement (1997-1998), respectively, we observed a marked decrease in the last period. Women and young people (under 30 years) experienced significantly higher absorption levels (respectively, 7.1% and 24.4%). CONCLUSION: The data suggest that monitoring was more frequent in subjects with higher starting values, and the greatest decrease was reported in this group. Reduction may be due to less n-hexane in the products used, better structural conditions in the factories, and the effectiveness of inspections carried out by the authority for hygiene and safety in the workplace. The results confirm the usefulness of the reporting of risk levels of exposure to industrial toxicants by routine biological monitoring. PMID- 12768282 TI - A 2-year follow-up survey of health and life style in Japanese unemployed persons. AB - OBJECTIVES: To clarify how unemployment affects health and life style in Japanese unemployed persons. METHODS: A group of 768 workers over 40 years of age in a large shoemaking company were dismissed because of sudden bankruptcy. Every 6 months after they had left the company, a survey with questionnaires was performed on 473 of them, who had organized an association. The questionnaires contained queries about somatic and mental symptoms and life style, including sleep, diet, exercise, smoking and drinking habits. The data from 263 of 473 subjects responding to four consecutive surveys every 6 months were analyzed in detail. RESULTS: The subjects kept their health and daily life style in good condition. This might have been due to the financial support of unemployment benefit. When the right to unemployment benefit expired, mood disorders occurred. In daily life, the prevalence rates of smoking and drinking consumption hardly changed in all subjects, and physical exercise was also habitually undertaken. The motivation for re-employment was remarkably enhanced by family economic crisis. CONCLUSION: Health, except for mental status, and life style of the unemployed persons were maintained in good condition. Mood disorders may result from prolonged jobless situations and cessation of the right to unemployment benefit. PMID- 12768283 TI - Exhaled NO level and number of eosinophils in nasal lavage as markers of pollen induced upper and lower airway inflammation in children sensitive to grass pollen. AB - OBJECTIVES: This study investigates the upper and lower inflammatory response induced by natural exposure to grass pollen in atopic and non-atopic children. METHODS: After children's atopic profile had been assessed, their nasal lavage fluid (NAL) and exhaled air was sampled once before and once during the pollen season. Level of nitric oxide (NO) was determined in exhaled air, and the following mediators were measured in NAL: ECP, IL-6, IL-8, albumin, uric acid, and urea. The number of eosinophils in NAL was determined after Giemsa staining. During the experiment ozone and pollen levels were measured continuously. RESULTS: During the pollen season the level of grass pollen was 95 pollen grains per cubic metre. At baseline, 8.0% and 5.4% of total cells in NAL of children sensitive to, respectively, house dust mite (HDM) and pollen + HDM were eosinophils, whereas virtually no eosinophils were observed in NAL of non-atopic children. In contrast to the non-atopic and HDM groups, in children sensitive only to grass pollen, grass pollen induced a threefold increase in the percentage of NAL eosinophils and a 2.5-fold increase in the NAL level of ECP ( P<0.05). In all groups, the NAL levels of albumin, uric acid, urea, IL-6 and IL-8 were not significantly increased by pollen exposure. At baseline, children sensitive to HDM showed significantly higher exhaled nitric oxide (eNO) values than non-atopic subjects and children sensitive only to pollen (79 to 141% increase). During pollen exposure eNO of children sensitive only to pollen increased from 35.8 to 64.5 ppb ( P<0.05), whereas no increase in eNO was observed in the other children. CONCLUSION: Pollen-sensitive children show a season-dependent upper and lower airway inflammatory response, resembling the continuous inflammation in HDM sensitive children. PMID- 12768284 TI - Neopterin as a new biomarker for the evaluation of occupational exposure to silica. AB - OBJECTIVES: Silica is one of the most documented workplace contaminants. Long term occupational exposure to silica is associated with an increased risk for respiratory diseases such as silicosis, tuberculosis, chronic bronchitis, chronic obstructive pulmonary disease and lung cancer. Furthermore, a variety of immune dysfunction-related diseases has been reported in silicotic individuals. Preliminary studies indicating enhanced levels of autoantibodies and several cytokines reflect an involvement of the immune system in the pathogenesis of silicosis and resulting complications. As an early and valuable biomarker of cellular immunity, neopterin is a low-molecular-mass compound belonging to the class of pteridines. It is produced by guanosine triphosphate via interferon gamma, following the activation of T cells. The aim of the present study was to observe the alteration of neopterin in silica exposure, and also to show whether screening of neopterin levels may be of use for assessment of occupational exposure to silica. METHODS: In this study, serum and urinary neopterin levels, both in silica-exposed workers ( n=22) and healthy volunteers ( n=20), were investigated by ELISA, spectrophotometry and HPLC techniques. RESULTS: Serum neopterin levels of control and exposed groups were measured as 5.98+ or -0.44 and 7.86+ or -1.97 nmol/l, respectively ( P<0.05). Urinary neopterin levels were also increased in the exposed group: 97.60+ or -41.42 micromol/mol creatinine for controls and 165.59+/-78.20 micromol/mol creatinine for workers ( P<0.05). At the same time, the correlation between urinary neopterin levels, serum neopterin concentration and working years, smoking status, some complaints, and silica status in the working atmosphere were evaluated. CONCLUSIONS: Our findings suggest that the following up of neopterin levels may have diagnostic value in silica-related diseases such as silicosis. Moreover, its biological monitoring should be performed in workplaces for clinical diagnosis and prognosis. PMID- 12768285 TI - Immunolocalization of the acid-sensing ion channel 2a in the rat cerebellum. AB - The acid-sensing ion channels (ASICs) are members of the DEG/ENaC superfamily of Na+ channels. Acid-gated cation currents have been detected in neurons from multiple regions of the brain including the cerebellum, but little is known about their molecular identity and function. Recently, one of ASICs (ASIC1a) was implicated in synaptic plasticity. In this study we examined the subcellular distribution of ASIC2a in rat cerebellum by immunostaining and confocal microscopy. Monoclonal antibodies for labeling of defined brain structures, for example, astroglia, Purkinje cell dendrites, nuclei, and presynaptic terminals were used for colocalization analyses. In the gray matter, the anti-ASIC2a antibody intensively stained dendrite branches of Purkinje cells evenly distributed throughout the entire molecular layer (ML). In the granule cell layer (GL), anti-ASIC2a antibody stained synaptic glomeruli. Neuronal localization of ASIC2a was confirmed by lack of co-staining with glial fibrillary acidic protein. Anti-ASIC2a staining in the ML colocalized with metabotropic glutamate receptor 1alpha (mGluR1alpha) in Purkinje cell dendrites and dendritic spines. Both proteins, mGluR1alpha and ASIC2a, were enriched in a crude synaptic membrane fraction prepared from cerebellum, suggesting synaptic expression of these proteins. Dual staining with anti-syntaxin 1A and anti-ASIC2a antibodies demonstrates characteristic complementary distribution of two proteins in both ML and GL. Because syntaxin 1A localized in presynaptic membranes and synaptic vesicles, complementary distribution with ASIC2a suggests postsynaptic localization of ASIC2a in these structures. This study shows specific localization of ASIC2a in both Purkinje and granule cell dendrites of the cerebellum and enrichment of ASIC2a in a crude cerebellar synaptic membrane fraction. The study is the first report of synaptic localization of ASIC2a in the CNS. The synaptic localization of ASIC2a in the cerebellum makes this channel a candidate for a role in motor coordination and learning. PMID- 12768287 TI - Villin is a possible marker of receptor cells in frog taste organs. AB - We investigated lingual taste organs of four frog species mainly by means of fluorescence immunohistochemistry for villin, calbindin, and serotonin. Cells immunoreactive for villin appeared in the taste organs of all the species used. These villin-immunostained cells were basoapically elongated in shape and extended up to the apical surface. They were also immunoreactive for calbindin. On the other hand, serotonin-immunoreactive cells, identified as Merkel-like basal cells, were immunonegative for villin. Considering the present results combined with those of studies by other research groups, the villin-immunostained cells were postulated to function as taste receptors. PMID- 12768286 TI - Effect of intestinal inflammation on capsaicin-sensitive afferents in the ileum of Schistosoma mansoni-infected mice. AB - In the present study, the effect of intestinal schistosomiasis on the extrinsic sensory innervation of the murine ileum was investigated. Immunocytochemical techniques to localize calcitonin gene-related peptide (CGRP), substance P (SP), and vanilloid receptor 1 (VR1) were combined with retrograde tracing techniques and capsaicin treatment. Neurochemical characterization of extrinsic primary afferent neurons (EPANs) in normal and capsaicin-treated mice, revealed that CGRP and VR1, but not SP, were expressed in extrinsic afferents. Immunocytochemical analysis using the above-mentioned antibodies yielded three different populations of neurons in both dorsal root and nodose ganglia, namely CGRP/--, SP/--, and CGRP/SP-expressing neurons. Retrograde tracing revealed that only CGRP/- expressing neurons projected to the ileum. Intestinal schistosomiasis resulted in an upregulation of the number of CGRP-immunoreactive (ir) nerve fibers in the lamina propria of the villi, coinciding with an increase in mucosal mast cells in acutely and chronically infected animals. In infected animals, mucosal mast cells were found closely associated with a dense mucosal CGRP-ir fiber network. Neonatal capsaicin treatment led to a 70% reduction in the number of mucosal mast cells. In conclusion, the present study provides evidence that CGRP is a valid marker for EPANs in the mouse ileum, which are involved in the recruitment of mucosal mast cells. Morphological evidence is provided of a neuroimmune interaction between mucosal mast cells and EPANs in schistosoma-infected mice. PMID- 12768288 TI - The change of etiological agents and clinical signs of epidemic viral conjunctivitis over an 18-year period in southern Taiwan. AB - BACKGROUND: Epidemic viral conjunctivitis is a highly contagious eye disease that occurs worldwide and is caused mainly by adenoviruses and enteroviruses. An 18 year analysis of the changes of pathogens and clinical signs in a subtropical and densely populated island presents certain special features. METHODS: We retrospectively analyzed the clinical information and laboratory records of the conjunctivitis patients with positive conjunctival swabs from 1980 to 1997. RESULTS: The positive rate of laboratory diagnosis of epidemic conjunctivitis was 50.0% (1,233/2,467). From 1980 to 1994, the predominant causative agent of adenoviral keratoconjunctivitis was adenovirus type 8 (Ad8), with six genotypes being evolved. Three of the new Ad8 genotypes each caused a new epidemic. After 1995 the predominant adenoviral pathogens shifted to Ad37 and Ad19, and no more Ad8 was isolated. Enterovirus type 70 (EV70) was isolated from four outbreaks of acute hemorrhagic conjunctivitis (AHC) from 1980 to 1984, but rarely in later years. Coxsackievirus A type 24 variant (CA24v), which first appeared in 1985, appeared later as the causes of four major epidemics of AHC from 1985 to 1994. The overall clinical symptoms of viral conjunctivitis were more severe in the 1990s than in the 1980s. CONCLUSION: In southern Taiwan, outbreaks of adenoviral keratoconjunctivitis caused by new genomic variants could be associated with the long-term endemic co-circulation of Ad8, Ad19, and Ad37, while epidemics of CA24v AHC were caused mainly by introduction of new viral strains from neighboring countries. The aggravation of host symptoms in the 1990s needs further investigation and close follow-up. PMID- 12768289 TI - Effect of linoleic acid and gamma-linolenic acid on tear production, tear clearance and on the ocular surface after photorefractive keratectomy. AB - BACKGROUND: The aim of this study was to evaluate the effect of linoleic acid (LA) and gamma-linolenic acid (GLA), both precursors of PGE(1), on tear production, tear fluorescein clearance and on the ocular surface after photorefractive keratectomy (PRK). METHODS: Sixty subjects (age 25+/-10 years; refractive error -3+/-2 diopters (spherical equivalent), mean +/- standard deviation) undergoing PRK were enrolled. The inclusion criteria were: Schirmer 1 test >10 mm/5 min, no corneal fluorescein staining, low irritation symptoms (questionnaire score <5), standardised visual scale (to evaluate tear fluorescein clearance) score <3. Patients were randomly divided into two groups. One group of 31 subjects was treated once daily orally with tablets containing LA (28.5 mg) and GLA (15.1 mg) (from 3 days before PRK to 1 month after PRK). The control group (29 subjects) underwent PRK and received no treatment with LA and GLA. A symptoms questionnaire, Schirmer 1 test and fluorescein clearance test (FCT) using the standardised visual scale were performed before starting therapy (T(0)) and 30 days after PRK (T(1)). RESULTS: All patients completed the study. The Schirmer 1 test varied from 16.3+/-6.9 (T(0)) to 17.6+/-7.2 (T(1)) for the treated group and from 18.3+/-6.2 (T(0)) to 15.7+/-7.4 (T(1)) for the untreated group ( P<0.0001, two-tailed unpaired t-test). FCT was 1.9+/-0.6 at T(0) and 1.6+/-0.8 at T(1) for the treated group and 1.7+/-0.7 at T(0) and 2.0+/-0.9 at T(1) for the untreated group ( P<0.0001). The symptoms score was 4.7+/-1.9 at T(0) and 7.6+/-7.2 at T(1) for the treated group and 4.2+/-2.0 at T(0) and 10.1+/ 7.6 at T(1) for the untreated group ( P<0.05). CONCLUSION: Reduced corneal sensitivity has already been proved after PRK. This could be the main reason for a decrease in tear production and for a reduced blinking rate leading to delayed tear clearance. The oral precursors of PGE(1), LA and GLA, could be helpful in increasing tear production and clearance after PRK. PMID- 12768290 TI - Extremely low frequency magnetic fields affect transposition activity in Escherichia coli. AB - The aim of this study was to verify whether extremely low frequency (ELF) magnetic fields (MF) could affect transposition activity like some environmental stress factors such as heat shock or UV irradiation. Using an Escherichia coli Lac Z(-) strain transformed with a plasmid containing a Tn 10 derivative element expressing beta-galactosidase only after transposition, it was possible to determine the events of transposition evaluating the rate at which the colonies developed dark coloured papillae (Lac Z(+)). We found that those bacteria that had been exposed for a long time (58 h) to a 50 Hz low intensity MF (0.1-1 mT) gave colonies with significantly lower transposition activity compared to sham exposed bacteria. Such reduction in transposition activity was positively correlated to the intensity of the MF, in a dose-effect manner. This phenomenon was not affected by bacterial cell proliferation, since no significant differences were observed in number, diameter and perimeter between sham-exposed and MF-exposed colonies. PMID- 12768291 TI - Changes in bone volume after irradiation with carbon ions. AB - The effects of carbon ions on bone volume were studied by histological and morphometrical methods. Anesthetized rats irradiated with three different single doses (15, 22.5, or 30 Gy) of either carbon ions or gamma rays were sacrificed (5 rats/group) at 10 time points together with non-irradiated controls. Carbon ion induced bone responses were qualitatively and quantitatively different from those induced by gamma irradiation at the same dose level. Irradiation with carbon ions resulted in a dose-dependent increase in bone volume, while gamma irradiation induced loss of bone volume, which was found to be independent of dose in the range studied over the same time course. After irradiation with carbon ions, bones showed thickening of the trabeculae, and the bone marrow became fibrous with fewer vessels. Carbon ion irradiation showed a greater effect on the bone absorbing capability of osteoclasts than gamma irradiation. These observations suggest that irradiation with carbon ions may produce differential modulation of radiation-induced growth factor expression. PMID- 12768292 TI - Lung cancer during pregnancy involving the products of conception and a review of the literature. AB - Approximately 1 per 1,000-1,500 pregnancies is complicated by maternal malignancies. Metastatic involvement of the products of conception is a rare event. There have been 62 cases of placental and/or fetal metastatic involvement originating from maternal cancer reported since 1866. Only 14 cases of lung cancer associated with pregnancy have been documented. We report on an additional case involving the products of conception, and the management of lung cancer in pregnancy is discussed based on an extensive review of the literature. The case of a 29-year-old woman presenting during the 31(st) week of gestation with metastatic non small-cell lung cancer to the placenta, liver and bone is described. The mother was delivered by caesarean section of a healthy baby girl during her 32(nd) week of gestation. The mother's postpartum course was complicated by disseminated pulmonary and bony metastases and malignant pericardial and pleural effusions causing the patient's death within 1 month after lung cancer was diagnosed. Malignancies spreading to the products of conception are melanoma (32%), leukemia and lymphomas (15%), breast cancer (13%), lung cancer (11%), sarcoma (8%), gastric cancer (3%) and gynecologic cancers (3%), reflecting malignancies with a high incidence in women of reproductive age. All lung cancers were diagnosed with widely disseminated, inoperable neoplastic disease, including distant metastases in 46%. The mean age was 35.1 years (range, 30-45 years) and 60% of patients had a history of tobacco use. The mean survival was 7.5 months (range: 1-42 months). Placenta involvement was present in 7 out of 15 cases. Fetal involvement was reported in only one case. Because there is no evidence of a direct adverse effect of pregnancy on the course of lung cancer, we recommend delivery at a time when enough fetal maturity can be assumed and the subsequent treatment of the mother. PMID- 12768293 TI - Expectations for success in fertility treatment involving assisted reproduction. AB - We carried out a prospective investigation recording how 115 couples estimated their chances for successful in-vitro fertilization. The couples were asked to predict the success of in-vitro fertilization on a scale ranging from 0% to 100%. The somatic findings and the social background variables (age, education, length of time of trying to get pregnant, causes of infertility and experience of treatment) were also recorded. Of these couples, 57% of the women and 61% of the men hoped for success. The expectations for success bore no relation to age, duration of infertility, education, experience with treatment or the causes of infertility. PMID- 12768294 TI - Frequency of glove perforation and the protective effect of double gloves in gynecological surgery. AB - The purposes of this prospective study were to verify the frequency of glove perforation during gynecological operations and to evaluate the efficacy of double gloving in preventing damage to the inner glove. From May 2000 to May 2001, three house staff and 12 residents were asked to place their used gloves in bags labeled with the following information: procedure performed, presence of a recognized glove perforation, and role in operating team (surgeon, first or second assistant, and instrumentalist). All glove sets were tested using the method of water pression. Damaged gloves were excluded from that analysis. In all, 35 and 51 operations were utilized with single and double gloves, respectively. There were 240 single gloves and 792 double gloves tested. Perforation occurred in 10.4% of the single gloves and 9.8% of the outer double gloves. There were no cases of perforation in the inner double gloves. In cases of operating time that lasted more than 2 h, 56% of the surgeries that used single gloves had perforation vs 58.5% of the double gloves. The first assistant had the major risk for glove perforation with the use of single or double gloves. The indicator finger of the non-dominant hand was the major risk for perforation. In conclusion, we recommend double gloving in all gynecological surgery to reduce the risk of contracting blood-borne diseases. PMID- 12768295 TI - Effects of short-time (3 months) tibolone treatment on bone turnover in postmenopausal women. AB - This study was planned to elucidate the effects of tibolone on bone biochemistry parameters in postmenopausal women at 3 month intervals. There were 56 healthy postmenopausal women enrolled in the study. The women had not received hormone replacement therapy (HRT) previously. Tibolone (2.5 mg/day) was prescribed for 3 months. Serum osteocalcin, calcium, phosphorus, alkaline phosphatase, creatinine and urine calcium, phosphorus creatinine, deoxypyridinoline were measured, and physical examinations were performed at the onset and at the end of the study. The mean serum osteocalcin level and deoxypyridinoline/creatinine (DPD/cr) ratio both decreased significantly (50.3% and 22.9%; P=0.012 and P=0.001, respectively). The slight decreases in serum alkaline phosphatase (4.5%) and urine calcium (13.6%) levels were not statistically significant. There was a positive correlation between DPD/cr and urine calcium ( r=0.66, P=0.001). We conclude that bone formation may be increased early by tibolone after short-term administration. PMID- 12768296 TI - The gynecologist's role in mammography screening in absence of a public health program. AB - This study examined the role of the gynecologist in a woman's adoption of mammography screening in a population that does not benefit from an organized program of breast cancer screening. We surveyed 486 women ages 50-69 years, randomly selected from the population of Geneva (Switzerland) about recent mammography use, attitudes toward mammography screening (decisional balance), whether they received a recommendation to undergo mammography from their gynecologist and/or family practitioner, visits to gynecologist and family practitioner, and sociodemographic variables. In multivariate analysis, on schedule mammography was associated with a recent visit to a gynecologist (odds ratio (OR) 4.8, 95% confidence interval (CI) 2.1-10.9), a gynecologist's recommendation for mammography (OR 15.8, 95% CI 6.5-38.6), a positive attitude toward mammography screening (positive score of decisional balance, OR 1.7, 95% CI 1.0-3.0), and being married (OR 2.0, 95% CI 1.2-3.3). Higher decisional balance scores were associated with a recent visit to a gynecologist ( P=0.001), a gynecologist's recommendation for mammography (P=0.001), and income ( P=0.01). Both a recent visit to a gynecologist and a gynecologist's recommendation for mammography were more frequent in women of higher income. This study suggests that gynecologists--but not family practitioners--play a key role both in referring women for mammography and in fashioning women's perceptions of mammography screening. Our data also indicate that a lack of access to a gynecologist and not receiving a gynecologist's recommendation for mammography may explain why low-income women are less likely to get a screening mammogram. PMID- 12768297 TI - The evaluation of cardiac biometry in major cardiac defects detected in early pregnancy. AB - The objective was to evaluate early cardiac biometry in fetuses with structural cardiac defects between 10 and 17 weeks of gestation using our normative data about fetal heart biometry. A retrospective case series, patients were selected from all cases with congenital heart disease diagnosed between 10 and 17 weeks of gestation in our prenatal unit between 1999 and 2000. A schematic sonographic examination, including nuchal translucency (NT) thickness measurements, was performed and was followed by fetal Doppler echocardiography. The transversal heart diameter, both ventricular dimensions, heart area, heart circumference, thoracic diameter, thoracic circumference, thoracic area, pulmonary trunk diameter and aortic diameter were measured and the cardiothoracic ratios were calculated. Doppler evaluation of the umbilical arteries, ductus venosus and umbilical vein was performed. Fetal karyotyping was obtained by amniocentesis or chorionic villous sampling. During the study period, 31 cases of congenital heart disease between 10 and 17 weeks of gestation were diagnosed. Of these, two fetuses presented with ectopia cordis and six with insufficient cardiac biometric measurements. In the remaining 23 fetuses, different complex abnormalities with a high rate of chromosomal abnormalities (91%) were present. Fetal heart biometry was normal in 22% and abnormal in 78%. NT thickness measurements were performed before 14 weeks of gestation and ten of 12 fetuses (83%) presented with an increased NT. Both fetuses with normal NT showed an abnormal fetal heart biometry. Venous Doppler evaluation was performed in 22 cases and 12 fetuses (55%) demonstrated an abnormal venous Doppler. There were ten fetuses (45%) with normal venous Doppler; in seven of these cases, fetal heart biometry was partly abnormal. This study shows the feasibility of first and early second trimesters' fetal echocardiography and the applicability of cardiac biometry in these instances. In this context, early fetal heart biometry and NT thickness measurements may be complementary methods for the prenatal diagnosis of some major congenital heart defects. In early pregnancy, some cardiac defects like tricuspid valve dysplasia, coarctation of the aorta, aortic stenosis, tetralogy of Fallot or pulmonary stenosis may already show similar changes in the relation of the diameters of the fetal heart and great arteries, as seen in the second trimester. Therefore, evaluating the different cardiac ratios may have a high diagnostic value in early pregnancy. PMID- 12768298 TI - Successful cerclage in two patients with advanced cervical dilatation in the second trimester. AB - Emergency cerclage in the second trimester is a recognized treatment for a dilatated cervix. Cervical cerclage is usually not attempted with advanced degrees of cervical dilatation because of the poor chances of survival. We had two rare cases of successful cerclage at advanced cervical dilatation in the second trimester and both delivered healthy babies at full term. PMID- 12768299 TI - Etidronate and hormone replacement therapy (HRT) for postmenopausal women with osteoporosis despite HRT. AB - Hormone replacement therapy (HRT) is effective treatment for postmenopausal bone loss and osteoporosis. However, postmenopausal women with low bone mass fail to respond after a few years of HRT and some postmenopausal women show no bone response to HRT. In these cases, additive therapy is necessary. Bisphosphonates are highly effective in postmenopausal osteoporosis. We investigated the effects of adding etidronate, a bisphosphonate, to HRT. The addition of etidronate to HRT improved bone density (4.40+/-1.12% increase per year) in patients showing no increase or even a decrease after at least 1 year of HRT (5.39+/-2.40% decrease per year). PMID- 12768301 TI - Detection of endothelial dysfunction in preeclamptic patients by using color Doppler sonography. AB - Our goal in this study was to detect endothelial function in normal and preeclamptic patients by using color Doppler sonography and plasma fibronectin levels. The increased ratio of the brachial artery diameter during shear stress, and plasma fibronectin levels were measured in 15 preeclamptic and 11 normal, gestational-age matched pregnant patients. The test was repeated at the postpartum second and sixth weeks in the preeclamptic group. In addition, the plasma fibronectin levels of all patients were measured. The increased ratios were 4.26+/-0.69% vs 12.18+/-1.97% in the preeclamptic and normal patients, respectively ( P=0.003). At the second and sixth postpartum weeks, the ratios were 6.67+/-0.89% and 9.27+/-1.16% in the preeclamptic group, revealing a significant improvement in the sixth week ( P=0.001). Fibronectin levels were 0.80+/-0.11 g/L vs 0.45+/-0.06 g/L in preeclamptic and normal patients ( P=0.01). The correlation coefficient between the fibronectin levels and increase rate was r=-0.38 and P=0.05. We conclude that endothelial dysfunction, which is fundamental to preeclampsia, can be detected by using color Doppler sonography. PMID- 12768300 TI - The comparison of clinical and hormonal parameters in PCOS patients treated with metformin and GnRH analogue. AB - In this study, our aim was to compare the effects of metformin and gonadotropin releasing hormone (GnRH) analogues in clinical and hormonal parameters in women with polycystic ovary syndrome (PCOS). There were 50 women with PCOS who were included in our study and they were divided into two groups. In this randomized trial, metformin (850 mg, two times per day) was administered to the first group and GnRH analogue (goserelin 3.6 mg, every 28 days) was given to the second group for 3 months. Because of the previous treatments, PCOS patients served as their own controls. The results of 42 women who completed the study were evaluated. Insulin resistance was not ascertained in patients. Metformin treatment resulted in a significant decline in mean body mass index, body weight, circumferences of waist and hip and total hirsutismus score. There was a significant decrease in luteinizing hormone (LH) levels and a significant increase in follicle stimulating hormone (FSH), progesterone, and sex hormone binding globulin (SHBG) concentrations. No changes in fasting glucose and insulin levels were observed. The GnRH analogue resulted in a significant increase in FSH and SHBG levels and a significant decrease in LH, total testosterone, dehydroepiandrosterone sulfate (DHEAS) levels and LH to FSH ratio and an improvement in hirsutism scores. Metformin and GnRH analogues had effects upon the abnormal steroid-gonadotropin metabolism and the clinical findings of hyperandrogenism with different mechanisms. PMID- 12768302 TI - Does consanguinity affect the severity of pre-eclampsia? AB - To determine whether consanguinity is more likely to be associated with severe forms of pre-eclampsia/eclampsia. Presuming a pure genetic contribution, we speculated that consanguineous marriages would increase the occurrence of severe forms of pre-eclampsia/eclampsia, through an expected increased chance for homozygosity to the putative gene. The study is a clinical case series on pre eclamptic/eclamptic primiparae delivered at Princess Badea Teaching Hospital, which is a tertiary referral center. The internationally accepted definitions for hypertension, proteinuria, mild and severe pre-eclampsia were adopted. The study included 77 primiparae. The incidence of consanguinity in the studied sample was 38%. Of them, 28 (36.4%) had mild pre-eclampsia, 45 (58.4%) had severe pre eclampsia and four (5.2%) had eclampsia. There was no statistically significant difference in the occurrence of severe pre-eclampsia/eclampsia between primiparae married to a first cousin or a relative other than a first cousin and primiparae married to a non-relative, odds ratio 1.1 (95% CI 0.33-3.87), P value 0.94, odds ratio 2.6 (95% CI 0.45-27.6), P value 0.30, respectively. Also, there was no statistically significant difference in occurrence of severe pre eclampsia/eclampsia between primiparae whose parents are first cousins or relatives other than a first cousin and primiparae whose parents are non relatives, odds ratio 1.3 (95% CI 0.36-4.72), P value 0.81, odds ratio 1.61 (95% CI 0.23-18.4), P value 0.70, respectively. Our study did not support a causal relationship between consanguinity and the occurrence of severe pre eclampsia/eclampsia. The role of more complex genetic, immunologic, metabolic, hemostatic or, possibly yet, other unknown factors have to be explored. PMID- 12768303 TI - Advantages of and applications for extended field-of-view ultrasound in obstetrics. AB - We evaluated the use of extended field-of-view (EFOV) ultrasound imaging in obstetrics and the possible advantages and applications. From July 2000 to May 2001, 57 randomly selected pregnancies and two woman postdelivery referred for routine and targeted ultrasound examination were studied by conventional B-mode and EFOV ultrasound. We subjectively compared conventional B-mode and EFOV by judging the general visualization as perceived by the operator and by the referring doctor. EFOV imaging provides better visualization of structures too large for a conventional sector image by creating one image of the entire structure and showing the relationship to or within the uterus. The EFOV images were easier to interpret by the referring or another nonexamining doctor. Fetal movements interfere with EFOV imaging, but not with the surrounding organs. A list of obstetrical applications for the use of EFOV is proposed. The use of EFOV imaging appears to be a reliable method to visualize the uterus in its entirety as well as genital tract tumors, the placental implantation and fetal structures throughout pregnancy. EFOV imaging of the fetus is limited by fetal movements. PMID- 12768304 TI - The primary varicella infection near term: a case report. AB - A 33-year-old pregnant woman developed skin rash at the 40 weeks' gestation. She was diagnosed as primary varicella zoster infection. Upon hospitalization, she was administered acyclovir and also a tocolytic agent (ritodorine hydrochloride) to postpone labor. The patient delivered a female baby 7 days after she developed skin lesions. Serological tests conducted after delivery showed that the mother was positive for IgG and IgM against varicella zoster. The baby was observed in a neonatal intensive care unit and found to be free of varicella zoster symptoms. PMID- 12768305 TI - Three-dimensional power Doppler ultrasound of the subendometrial blood flow under the administration of a contrast agent (Levovist). AB - METHODS: Based on the results from 42 IVF patients on the day of oocyte aspiration, our study tested the hypothesis that 3-D Doppler ultrasound of the endometrium supported by a contrast agent possesses advantages over a conventional 3-D Doppler ultrasound examination. We also determined whether the subendometrial blood flow indices measured under continuous infusion of Levovist (Schering, Berlin, Germany) were an indicator of a successful implantation. RESULTS: It was demonstrated that, after the administration of Levovist, all subendometrial blood flow parameters (e.g., vascularization index [VI: 8.82+/ 16.64], flow index [FI: 35.67+/-9.33], and vascularization flow index [VFI: 3.99+/-8.80]) were significantly higher than without the contrast agent ( p<0.001; VI: 3.09+/-7.11; FI: 29.13+/-14.26; VFI: 1.35+/-3.31). However, no significant differences between blood flow with contrast agent and pregnancy rate were found. CONCLUSION: In summary, quantitative 3-D Doppler ultrasound employing Levovist (Schering, Berlin, Germany) is significantly more sensitive when compared to conventional 3-D Doppler examination. For the first time this was demonstrated on 42 IVF patients on the day of oocyte aspiration. However, no correlation with outcome of the IVF-treatment could be found. PMID- 12768306 TI - Ovarian metastases caused by cholangiocarcinoma: a rare Krukenberg's tumour simulating a primary neoplasm of the ovary: a two-case study. AB - INTRODUCTION: Ovarian metastases represent 10% of all ovarian malignancies. They are characterized by their ability to occasionally reproduce mimicking the clinical and morphological appearance of primary tumours, making diagnosis difficult. Most of these tumours originate in the digestive tract, the best known being Krukenberg's tumour in the stomach. Cholangiocarcinomas are rare neoplasias that very rarely affect the ovary. Their retroperitoneal location and low histological specificity in the ovary may lead to a misdiagnosis. This makes it necessary to accurately assess all clinical, radiological and morphological symptoms so as to avoid inappropriate treatments in affected patients. CASE REPORT: Two cases of biliary cholangiocarcinomas simulating an ovarian neoplasia are reported. PMID- 12768307 TI - Proliferation-differentiation relationships in the expression of heparin-binding epidermal growth factor-related factors and erbB receptors by normal and psoriatic human keratinocytes. AB - The topological relationships between erbB receptors and ligands of the epidermal growth factor family were characterized by immunocytochemistry in normal and psoriatic epidermis and in proliferating and differentiating human keratinocytes in culture. Spatial colocalization of receptors and ligands was assessed by dual immunostaining. Expression of epidermal growth factor receptor (EGFr), erbB2, and erbB3, but not erbB4, was detected throughout the epidermis, although labeling for erbB2 and erbB3 was accentuated in the upper spinous layers, and EGFr was more strongly labeled in basal cells. Of the tested growth factors, heparin binding epidermal growth factor (HB-EGF) was diffusely expressed throughout normal and psoriatic epidermis and sparsely colocalized with EGFr in all viable epidermal layers, with increased colocalization in psoriatic epidermis. In contrast, betacellulin and heregulin/neu differentiation factor (NDF) alpha were largely restricted in their distribution to the upper spinous and granular layers. Betacellulin was downregulated in psoriatic keratinocytes. Although heregulin/NDF-beta was undetectable in normal epidermis, it was upregulated in psoriasis. Betacellulin and heregulin/NDF-alpha strikingly colocalized with EGFr and erbB3 receptors in the granular layer and in a declining gradient from the granular zone to the basal layer, respectively. Similar patterns were observed in cultured keratinocytes under proliferative conditions and upon differentiation in high-calcium medium. These morphological data collectively suggest divergent functions for members of the growth factor family, and in particular, we propose that betacellulin and heregulin/NDF-alpha are involved in epidermal morphogenesis and/or in maintenance of the differentiated phenotype. PMID- 12768308 TI - Posttraumatic contracture of elbow treated with intraarticular technique. AB - BACKGROUND: Posttraumatic contracture of the elbow (either flexion or extension) is sometimes very disabling. However, an absolutely convincing surgical technique has not yet been defined in the literature. We developed an intraarticular technique to concomitantly treat both intraarticular and extraarticular lesions with one posterior incision. METHODS: Twenty consecutive adult patients were treated. After the olecranon was osteotomized, all intraarticular pathologies and the anteroposterior capsule were corrected completely. The olecranon was then stabilized with the modified tension band wiring technique. Immediately postoperatively, continuous passive movement was performed, and range-of-motion exercise of the elbow was encouraged continuously. RESULTS: All 20 patients were followed up for a median of 3.8 (range 2.1-6.6) years. The satisfactory rate was 95% (19 of 20, p<0.001). The flexion contracture improved from an average of 42 to 13 deg (p<0.001), and the maximal flexion improved from an average of 89 to 131 deg (p<0.001). The arc of motion improved from an average of 47 to 118 deg (p<0.001). The sole unsatisfactory patient still had 20-110 deg of arc of motion. There were no evident complications noted. CONCLUSION: Compared with other techniques, we recommend this one due to its high satisfactory rate and low complication rate. PMID- 12768309 TI - The Artek cup for total hip replacement of dysplastic hip joints. AB - BACKGROUND: One specific difficulty in total hip replacement for developmental dysplasia of the hip is a shallow acetabulum with a short anteroposterior diameter. In a prospective cohort study we investigated the short-term results of the Artek cup, designed with a shallow outline and a height-reduced metal inlay for a large size 38 mm metal head, in a group of female patients with mild dysplasia of the hip. METHODS: Fourteen consecutive patients with 17 dysplastic hip joints were included in the study. Their average age was 42 years. Fourteen hips had dysplasia Crowe grade I; 3 hips had dysplasia Crowe grade II. The femoral head centre was localized according to Pagnano in zone 1 in 3 cases, in zone 3 in 6 cases, and in zone 4 in 8 cases. The Harris hip score was 49 points (range 30-68 points). RESULTS: Mean follow-up was 20 months (range 14-24 months). The Harris hip score improved to a mean of 91 points (range 78-98 points). The postoperative radiographic measurements showed an inclination angle of 52.1 degrees (range 43-66 degrees ). The femoral head centre was in Pagnano zone 1 in 13 cases, in zone 2 in 1 case, in zone 3 in 1 case, and in zone 4 in 2 cases. The mean postoperative deviation from the approximative rotational centre was 0.17 mm (range -15 to 13 cm) in the superior direction and 5.3 mm (range -5 to 17 mm) in the lateral direction. CONCLUSION: In a small series of 17 implantations, stable press-fit fixation without screws or bone grafting was achieved despite acetabular defects. The complication rates were high. Inclination angles up to 54 degrees were not correlated with complications. Inclination angles of 60 degrees and more led to subtotal or complete dislocation. Mid-term results are still to follow concerning wear, dislocation rate, loosening and revision. PMID- 12768311 TI - Three-dimensional image reconstruction of an anorectal malformation with multidetector-row helical computed tomography technology. AB - The presentation of the surgical anatomy of anorectal malformation by standard anatomical figures is not suitable for individual anorectoplasty. It is essential to understand the anatomy of the pelvic muscle (striated muscle complex: SMC) including the external anal sphincter and their three-dimensional (3D) configuration in each patient. Thus, we studied the SMC three-dimensionally with multidetector-row helical computed tomography (MRH-CT) preoperatively, and evaluated its usefulness. Fourteen patients with anorectal malformations before anorectoplasty (types: high n=6, intermediate n=2, low n=6) and two patients without anorectal malformations as controls (total: male n=8, female n=8) were investigated. An image of pelvic region was prepared with a slice thickness of 0.5 mm and a reconstruction pitch of 0.5 mm. A 3D reconstruction on a conventional personal computer (PC) was made with a volume rendering method, and assisted by our own software. The SMC was analyzed with three modified modes of 3D reconstruction corresponding to the surrounding tissues. A length of the parasagittal muscle, and both the sagittal and transverse width of the vertical fibers in the SMC at the connection to the parasagittal muscle were measured on a 3D image and then compared among three different types and controls. To eliminate variations in age, a length index was used to allow comparison. The 3D configuration of the SMC was different in every case. The arranged image mode, which displayed the SMC and the pelvic bones simultaneously, enabled to use conventional knowledge in cysto-urethrography. The length of the parasagittal muscle was longest in the high type but the width of the vertical fibers was smallest. Anatomical figures of the SMC including the external anal sphincter were clearly demonstrated on a PC in every anorectal malformation by our program. A 3D reconstruction image provides positional information on the SMC for the body surface and pelvic bone at the same time. Both a 3D image and positional information with MRH-CT offers the surgeon a simulated operative profile of the SMC superior to CT or magnetic resonance imaging slices alone. PMID- 12768312 TI - Experience in 100 cases with the Nuss procedure using a technique for stabilization of the pectus bar. AB - The Nuss procedure is a new, minimally invasive technique for the repair of pectus excavatum. We describe our experience with this operation, in a relatively large series in a single institution, and introduce our technical modification for stabilizing the pectus bar. In 107 patients (75 male and 32 female) with pectus excavatum who underwent the Nuss procedure, age at operation ranged from 3 to 23 years old (mean: 7.5+/-4.1) and Haller's CT index ranged from 33 to 2.6 (mean: 6.1+/-3.5). To stabilize the pectus bars, we tied the rib and both edges of the bars with surgical steel wire, 0.8 mm in diameter, using a Duchenne needle to guide the wire behind the rib. Average operating time with this technique (n=100) was 48+/-20 min. The stability of pectus bars after the operation was assessed by lateral chest X-ray films. The position of the center of the pectus bars, facing the sternum at right angles, was classified as excellent. A minimal bar displacement of less than 45 degrees was classified as incomplete. The position of the bars that had rotated 90 degrees was classified as poor. The post operative course was uneventful in all cases except for three patients who showed wound infection, hemothorax or pneumothorax, independently. Two patients required wire removal due to skin irritation as a late complication. Five patients underwent re-operations due to bar displacement. Among them, only two cases were included in the group of wire fixation. Only five teen-aged patients required a lateral stabilizing bar. Lateral chest X-ray films of 100 patients showed that the position of the pectus bar was excellent in 86 cases, incomplete in 12 cases and poor in two cases. In 70 patients aged less than 10 years, the position in 91% was excellent and in 9% was incomplete. In 30 patients aged 10 years and older, 14 required two pectus bars and the results were excellent in all cases. The other 16 patients with a single bar showed excellent results in eight (50%) cases, incomplete in six (37.5%) and poor in two (12.5%). The wire-fixation technique was safe, effective and time efficient to perform in young cases. Teen aged patients, however, were at high risk of bar displacement. Therefore, the use of two bars and lateral stabilizing bars are recommended for these high-risk patients. PMID- 12768313 TI - Drainage of subcutaneous lymphatic fluid for the management of respiratory distress in a case of generalized lymphangiectasia in an infant. AB - A 10-month-old girl was referred to our hospital because of congenital and persistent bilateral chylothorax and generalized lymphedema as well as long standing respiratory disturbance. Radiological studies showed a diffuse network of superficial lymphatic vessels without major trunks throughout her entire body as well as the lung. She was diagnosed with systemic lymphangiomatosis complicated with pulmonary lymphangiectasia. Percutaneous puncture in the lower leg was performed to discharge the lymphatic fluid and proved to be effective for the respiratory disturbance. This procedure is safe and easy and effectively improves the quality of life of the patient and the family in case of such a persistent disease. PMID- 12768314 TI - Analysis of treatment outcome for children with recurrent or metastatic hepatoblastoma. AB - For better total survival rate of children with hepatoblastoma, the therapeutic strategy for recurrent or metastatic hepatoblastoma should be improved. From 1991 to 1999, 134 cases of hepatoblastoma were treated by surgery and combination chemotherapy of cisplatin (CDDP) and THP-Adriamycin (THP-ADR) based on the JPLT-1 protocol. In 114 non-metastatic cases, 90 primary liver tumors were resected completely by partial hepatectomy, but 12 recurrences were observed in the liver (4 cases) and the lungs (8 cases). Distant metastases on the diagnosis were observed in 20 cases. The treatment outcome of these 12 recurrent and 20 metastatic tumors was analyzed. In four recurrent liver tumors, surgical resection was performed in all four cases, and all the patients were alive and well. In eight recurrent lung tumors, surgical resection was performed completely in six cases with unilateral lung disease, and five of the six patients were alive and well. In stage IV tumors, the survival rate of the patients having primary tumors within two hepatic sections was significantly higher than that of the patients having primary tumors over three hepatic sections. Active surgical intervention to lung metastases and a more intensive chemotherapy to facilitate complete resection of primary hepatic tumor could improve survival rate of children with refractory hepatoblastoma. PMID- 12768315 TI - A case of ischemic jejunal stricture after surgical reduction of intussusception. AB - A 15-year-old girl presented with small bowel obstruction due to ischemic jejunal stricture which developed three weeks after successful surgical reduction of an intussusception with a Peutz-Jeghers-type polyp as a lead point. The reduced jejunum had no macroscopic injury, and the stricture caused complete obstruction requiring jejunal resection. PMID- 12768316 TI - A case of neonatal cerebellar abscess. AB - CASE REPORT: A 1-month-old boy presented with repeated vomiting without signs of infection and was eventually found to have staphylococcal cerebellar abscess. The initial diagnosis was hypertrophic pyloric stenosis. RESULT: . He was successfully treated by drainage surgery combined with intravenous antibiotic therapy. CONCLUSIONS: . Neonatal brain abscess, particularly cerebellar abscess, is seldom encountered, but may cause disability in later years even after successful treatment. Early diagnosis and appropriate treatment are mandatory for these patients. PMID- 12768318 TI - Experience with different questionnaires in the management of patients with CP/CPPS: GPSS, IPSS and NIH-CPSI. AB - A number of different self assessment questionnaires have been developed in order to evaluate specific symptoms of chronic prostatitis. The most popular indices in Germany are the Giessen Prostatitis Symptom Score(GPSS), the International Prostate Symptom Score (IPSS) and the Chronic Prostatitis Symptom Index of the National Institutes of Health (NIH-CPSI). The major aim of our study was the evaluation of these questionnaires in patients with chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS). In addition, we analysed questionable differences in symptomatology between the subgroups NIH III A and B. A total of 203 native speaking German men with symptoms of CP/CPPS attending the Giessen prostatitis outpatient department were included in our protocol. According to the strict criteria of the Giessen examination protocol, 84 men were classified as CP/CPPS type NIH III B and 54 men as NIH III A. The psychometric and descriptive results were analysed with SPSS software. The Cronbach alpha, as a parameter for the internal consistency for each index, showed acceptable values. The correlation coefficient for each index was also sufficient. The median total scores of the GPSS and NIH-CPSI were significantly higher in patients with CP/CPPS type NIH III B. The IPSS results were the same. All symptom indices demonstrated acceptable to good values for psychometric validation. Similar symptomatic findings were available using GPSS and NIH-CPSI. PMID- 12768317 TI - Craniocerebral gunshot injuries in children. AB - INTRODUCTION: Despite the worldwide increase in the incidence of gunshot injuries, there are few large published series on craniocerebral gunshot injuries in children. MATERIALS AND METHODS: The records of 30 consecutive children who were treated for craniocerebral gunshot injuries at the Red Cross War Memorial Children's Hospital from 1989 to 2002 were reviewed retrospectively. The circumstances of the injury, clinical status, CT findings, complications, and outcome were assessed. RESULTS: The median age was 7 years. Seventy-seven percent of the victims were boys. The majority of the children were injured in the crossfire of civilian violence. The initial management consisted of debridement under local anesthesia in 16 children and neurosurgical procedures under general anesthesia were performed in 14. Sixteen children sustained transhemispheric injuries, 5 bihemispheric injuries, 5 tangential injuries, and 4 transventricular injuries. All 3 children with a GCS <4 died within 72 h of admission. Three of the 7 children with GCS 4-7 died but there were no deaths in those children whose GCS was >7 post-resuscitation. Motor deficits, cranial nerve palsies, and visual field defects were very common. Early post-traumatic seizures were the commonest complication (18%). CONCLUSION: Children with higher post-resuscitation GCSs fared better than adults in terms of mortality but not necessarily morbidity. As in the case with adults, the GCS after resuscitation is a very good prognostic indicator of mortality. PMID- 12768319 TI - Modelling of ftorafur and 5-fluorouracil pharmacokinetics following oral UFT administration. A population study in 30 patients with advanced breast cancer. AB - PURPOSE: The pharmacokinetics of ftorafur, 5-fluorouracil (5FU) and uracil were investigated in order to built a population pharmacokinetic model for the anticancer drug UFT, administered with leucovorin and vinorelbine. METHODS: A total of 31 patients with metastatic breast cancer were treated with escalating oral doses of UFT (300 to 500 mg per day) plus leucovorin (90 mg per day) in combination with intravenous vinorelbine (15 to 25 mg/m(2)). Concentration-time data were obtained on days 1, 8, 15 and 21 of cycle 1. RESULTS: Of the 31 patients treated, 30 were available for the pharmacokinetic analysis. Ftorafur, 5FU and uracil appeared rapidly in plasma and showed large interpatient variations. Ftorafur concentrations were higher than those of 5FU and uracil. AUC significantly increased between day 1, and days 8, 15 and 21. Ftorafur C(max) and AUC values were proportional to UFT dose, whereas C(max) and AUC values of 5FU and uracil were not linearly related to UFT dose. The pharmacokinetics of ftorafur were ascribed to a two-compartment open model in which 5FU was produced from the central compartment. The absorption and exponential distribution rate constants were assumed equal. The effect of uracil on 5FU elimination was straightforward, since no reasonable curve-fitting could be obtained for 5FU data when this covariate was not taken into account. The uracil concentration inducing a 50% reduction in 5FU elimination was 2.67 micro mol.l(-1). This result confirms the important role played by uracil as a competitive inhibitor of 5FU catabolism. CONCLUSION: A pharmacokinetic model for ftorafur and 5FU was developed and should be useful to further study drug interactions and establish dosing guidelines. PMID- 12768320 TI - A phase I trial of weekly paclitaxel, 13- cis-retinoic acid, and interferon alpha in patients with prostate cancer and other advanced malignancies. AB - PURPOSE: Based on prior studies demonstrating the effect of 13- cis-retinoic acid and interferon alpha (CRA/IFN) in decreasing the expression of the antiapoptotic protein bcl-2, our prior clinical study of CRA/IFN with paclitaxel (TAX) administered every 3 weeks, and data demonstrating increased activity of weekly TAX against prostate cancer, we designed a phase I study of weekly TAX in combination with CRA/IFN in patients with prostate cancer and other advanced malignancies. To develop a marker of drug effect, we assessed bcl-2 downregulation in patient peripheral blood mononuclear cells (PBMC). METHODS: Enrolled in the study were 14 patients with prostate cancer or other advanced malignancies, and 13 were treated with 1 mg/kg CRA on days 1 and 2, 6 MU/m(2) IFN subcutaneously on days 1 and 2, and TAX at increasing doses on day 2 each week for 6 weeks out of an 8-week cycle. The effect of CRA/IFN on bcl-2 expression was assessed in PBMCs by immunoblotting. RESULTS: The combination of CRA/IFN and TAX was well tolerated. Dose-limiting toxicities (DLT) in the first cycle of therapy included one patient with fever and neutropenia, and one patient with grade 4 hypertriglyceridemia. The recommended phase II dose of TAX in this combination was 80 mg/m(2). Of 13 patients assessable by tumor markers or scans, 5 had stable disease and 2 had a biochemical partial response including a patient with a decrease in PSA of >50% while on study. The assessment of patient PBMC bcl-2 was feasible in ten patients. CONCLUSIONS: This is the first study in which the safety and clinical activity of weekly TAX combined with CRA/IFN has been demonstrated. The assessment of PBMC bcl-2 is feasible in this weekly chemotherapy schedule PMID- 12768321 TI - Quantitative analysis of mixed chimerism following allogeneic bone marrow transplantation using telomere flow-FISH. AB - Assessment of mixed chimerism is of particular interest for allogeneic bone marrow or peripheral blood stem cell transplantation with reduced-intensity conditioning in order to study contribution of donor-type and host-type lymphohematopoiesis. Because the length of telomere repeat sequences is frequently shorter in leukemic compared to normal hematopoietic cells, this telomere repeat polymorphism might be a useful marker to analyze mixed chimerism in selected patients with short telomeres. Recently, fluorescence in situ hybridization and flow cytometry (flow-FISH) have been shown to be valuable tools to analyze the mean telomere length in hematopoietic cells. Here, we demonstrate in a case study on a patient with chronic lymphocytic leukemia (CLL) that telomere flow-FISH can in principle be exploited to quantitate the amount of donor- and host-type cells for chimerism analysis based on distinct histogram distributions which reflect cell populations with different telomere length. PMID- 12768322 TI - Primary extramedullary myeloid tumor of the breast: a case report and review of the literature. AB - Primary extramedullary myeloid tumors (PEMMT) are extramedullary proliferations of myeloid cells occurring in the absence of an antecedent myeloproliferative disorder. They have predilection for the skin, lymph nodes, central nervous system, and small intestine. Breast is an uncommon location for PEMMT and only a few cases have been reported so far in the medical literature. Many of these cases have been initially misdiagnosed. We reviewed all the reported cases of PEMMT of the breast in the English-language literature since 1965. In addition, we present a new case of PEMMT of the breast who presented with a mass in her right breast mimicking a breast malignancy and was initially misdiagnosed as non Hodgkin's lymphoma. A careful histologic examination with immunohistochemical studies revealed the presence of PEMMT of the breast. Treatment with systemic chemotherapy and local radiotherapy rendered a complete remission. Seventeen cases of PEMMT of the breast were reported in the English literature from 1965 to 2003. Most of the cases were misdiagnosed initially as lymphomas, breast carcinomas, or malignant melanomas. PEMMT of the breast is a poorly recognized entity whose diagnosis frequently challenges both the pathologist and the oncologist. Given the small number of patients reported no optimal treatment has been defined, but systemic chemotherapy similar to that given for acute myeloid leukemia with or without local radiotherapy may result in long remissions and avoid the progression to overt acute myeloid leukemia. PMID- 12768323 TI - Prenatal diagnosis in hemophilia A using factor VIII gene polymorphism--Indian experience. AB - The heterogeneous nature of the mutations, the size, and the complexity of the factor VIII gene makes direct mutation analysis in hemophilia A families in India an option that is not very feasible and practical. Thus, carrier screening and prenatal diagnosis of hemophilia A often depends on haplotype analysis using restriction fragment length polymorphisms (RFLP) and short tandem repeat (STR) markers to track the defective factor VIII gene within a family. The main objective of this present study was to assess the utility of using polymerase chain reaction (PCR)-based five polymorphic markers: four intragenic Hind III, Bcl I, intron 13, and intron 22 STRs and one extragenic marker St14 in prenatal diagnosis. Forty-one chorionic villus samples (CVS) were studied from 41 families with a history of hemophilia A. PCR and RFLP were used for screening. Intron 22 STR showed the highest informativeness (60.9%), followed by Hind III (51.2%), Bcl I (46.3%), & intron 13 STR (51.2%); the extragenic marker St14 (VNTR) was informative in 46.3% of families. Linkage analysis, with the combined use of these five PCR-based polymorphic markers, gives good informativeness of 87.8% in the Indian population. Of the 41 CVS tested, 21 were found to be male fetuses and of these 13 were found likely to be affected with hemophilia A. Only in 12.2% of the families were none of the markers informative. PMID- 12768324 TI - Primary aleukemic myeloid leukemia cutis treated successfully with combination chemotherapy: report of a case and review of the literature. AB - Aleukemic myeloid leukemia cutis is extremely rare and is usually associated with early marrow relapse and poor treatment outcome. We report a 39-year-old man presenting with generalized cutaneous nodules. The initial diagnosis was cutaneous malignant lymphoma. New skin lesions and a nasopharyngeal mass developed during phototherapy. Biopsy of the cutaneous and nasopharyngeal lesions revealed monotonous blast cell infiltration. Cytochemical stain and immunophenotypic analysis of the fresh cell suspension made from another skin biopsy specimen identified that the neoplastic cells belonged to the monocytic lineage. A diagnosis of primary aleukemic leukemia cutis was established. The leukemic cells expressed CD56 but did not carry AML-1/ETO, CBFbeta/MYH11, or common MLL fusion transcripts. He received standard induction therapy for acute myeloid leukemia, followed by high-dose postremission chemotherapy and has been disease-free for more than 30 months. To the best of our knowledge, the current case has the longest disease-free survival among those reported. PMID- 12768325 TI - A very rare anatomic variation of the left brachiocephalic vein: left retro aortic brachiocephalic vein with tetralogy of Fallot. AB - An anomalous course of the left brachiocephalic vein behind the aortic arch was identified in a patient with tetralogy of Fallot. A 6-year-old male patient had been admitted to the hospital with a diagnosis of tetralogy of Fallot and patent foramen ovale. At the beginning of the operation the left brachiocephalic vein could not be found. The pericardium was opened longitudinally and there was no persistent left superior vena cava. After careful examination, the left brachiocephalic vein was found behind the aorta. Especially during open heart surgery this pathology is very important. We conclude that when the left brachiocephalic vein can not be found, the possibility of persistent left superior vena cava and retro-aortic left brachiocephalic vein should be borne in mind. PMID- 12768326 TI - Stabilization of the ulna stump after Darrach's procedure at the wrist. AB - We studied the outcome of patients with chronic distal radioulnar joint symptoms who were treated with excision of the distal ulna and reconstruction using the flexor carpi ulnaris tendon. Twelve patients with 14 wrists were assessed. Ten patients were posttraumatic, and two had bilateral surgery for rheumatoid arthritis. Average age was 37 years at time of operation. Follow-up averaged 20 months (9 months to 4 years). All patients except one reported improvement in pain symptoms. Grip strength of the operated hand as measured by dynamometer readings was 67% of the strength of the normal hand with five wrists achieving an excellent result in grip strength. Eleven patients reported a subjective improvement in functional activities. An improved range of motion was obtained in all patients. We discuss the importance and basis for ligamentous reconstruction following excision of the distal ulna and review the literature for other previously described procedures. PMID- 12768328 TI - Intralesional lipid-complexed cytokine/superantigen immunogene therapy for spontaneous canine tumors. AB - These studies sought to determine the gene expression and short-term effects of intralesional lipid-complexed immunogene therapy with constructs encoding Staphylococcus aureus enterotoxin A and canine interleukin-2 (L-SEA/cIL-2) in dogs with tumors of various histotypes, and then to assess the safety and efficacy of repeated L-SEA/cIL-2 injections in dogs with spontaneous soft tissue sarcomas (STS). In the first study, pet dogs with a variety of tumors received a single intralesional injection of L-SEA/cIL-2, and surgical excision was performed 48 h later. In the second study, dogs with histologically confirmed STS were treated weekly for a maximum of 12 weeks with escalating doses of L-SEA/cIL 2. Tumors were then surgically excised and assessed histologically and immunohistochemically. Overall, treatments were well tolerated, with no dose limiting toxicities encountered. At 48 h, in the single injection study, plasmid DNA was detected in 14 of 16 tumor samples, and plasmid-specific mRNA was detected in 3 of 14. In the multiple injection study, the overall response rate in dogs with STS was 25%, consisting of 3 complete responses (CR) and 1 partial response (PR). Diffuse lymphoplasmacytic inflammation was observed in all tumors from patients experiencing CR or PR, whereas these changes were not evident in tumors from nonresponders. The infiltrate was composed primarily of CD3(+) cells at 48 h from the single-injection study, and was composed of both CD3(+) and CD79a(+) cells at 12 weeks in responding dogs from the multiple-injection study. In conclusion, these studies suggests that intralesional L-SEA/cIL-2 immunotherapy is well tolerated, results in detectable transgene expression in canine tumors, and has antitumor activity in dogs with spontaneous STS. PMID- 12768327 TI - Targeted cytotoxic effect of anti-JL1 immunotoxin against a human leukemic cell line and its clinical implications. AB - We have previously reported the identification of a unique thymocyte-specific surface molecule, JL1, which was detected using the monoclonal antibody (mAb), anti-JL1. Interestingly, JL1 was shown to be expressed in most leukemias, irrespective of their immunophenotype, and subpopulations of normal bone marrow (BM) mononuclear cells (MNCs). Here we investigated the potential usefulness of the anti-JL1 mAb as a therapeutic tool for leukemia. We demonstrated that the proliferation of cultured human leukemia cells was dramatically inhibited in vitro by anti-JL1 mAb conjugated with the polypeptide toxin, gelonin, but not by gelonin alone. We then systematically investigated the reactivity of the anti-JL1 mAb against normal human tissues to evaluate possible side effects along with various hematopoietic and nonhematopoietic tumor cell lines. All of 33 types of normal tissues except thymus and subpopulation of BM MNCs were clearly devoid of JL1 expression. Among tumor cell lines, all the nonhematopoietic cell lines tested were negative for JL1 expression, while some hematopoietic cell lines contained JL1 antigen. Collectively, the results showed the cytotoxic effects of anti-JL1-based immunotoxin against JL1-positive leukemic cells, sparing most normal tissues other than thymocytes and some BM MNCs. Therefore, we strongly suggest that gelonin-conjugated anti-JL1 mAb immunotoxin could be developed as a potential immunotherapeutic agent in the treatment of various types of JL1 positive acute leukemias. PMID- 12768330 TI - Improved kinetic stability of DTPA- dGlu as compared with conventional monofunctional DTPA in chelating indium and yttrium: preclinical and initial clinical evaluation of radiometal labelled minigastrin derivatives. AB - The development of monofunctional DTPA derivatives has been a major breakthrough in the labelling of proteins or peptides with a variety of radiometals. Although this methodology is simple and useful for indium-111 labelling, the stability of these conjugates is too low for most therapeutic nuclides. Cyclic chelators, such as DOTA, have shown excellent kinetic stability with a variety of radiometals, but the labelling procedure is more difficult, requiring ultra-pure reagents and a heating step that sometimes endangers the biomolecule's integrity. The aim of this work was twofold: (a) to develop a novel, open chain chelator which can be easily labelled with various radiometals, displaying higher kinetic stability than monofunctional DTPA, and (b) to evaluate this chelator in vitro and in vivo when conjugated to a CCK-B receptor ligand as a detection modality for receptor (over-)expressing tumours. DTPA derivatives of Leu(1)- and dGlu(1)-minigastrin were synthesised. All conjugates could be labelled with (111)In or (88/90)Y at high specific activities (8.5-44.4 GBq/micro mol) and with high radiochemical purity. Serum stability testing was performed, and the labelled conjugates were compared concerning their stability against DTPA challenge. The biodistribution of the radiolabelled Leu(1)- and dGlu(1)-minigastrin derivatives was studied in tumour-bearing nude mice, in one healthy human volunteer and in three patients with metastatic medullary thyroid carcinoma. The transchelation of all tested radiometals to serum proteins was significantly slower with the DTPA-Glu conjugates as compared with their Leu analogues (e.g. transchelation t(1/2) of DTPA- dGlu(1)-minigastrin vs its Leu(1) analogue at 37 degrees C in human serum for (111)In: 239 h vs 91 h; for (90)Y: 130 h vs 53 h). In animals, all labelled CCK-B receptor ligands showed fast and specific uptake in CCK-B-receptor-positive tissues, such as the stomach and tumour, as well as a fast renal clearance pattern. However, DTPA-Leu(1)-minigastrin showed higher background activity in the whole body and those organs known to accumulate the respective free radiometal (e.g. (88)Y-DTPA-Leu(1)-minigastrin had bone uptake of 22%ID/g as compared to only 1.2%ID/g with its dGlu(1) analogue). In humans, fast tumour and stomach uptake was observed for both (111)In-labelled compounds, but DTPA- dGlu(1)-minigastrin lacked the liver, spleen and bone marrow uptake observed with its Leu(1) analogue. In conclusion, anionic amino acid derivatives of DTPA may display improved metabolic stability as compared with monofunctional DTPA conjugates. DTPA- dGlu(1)-minigastrin is preferred to "monofunctional" DTPA Leu(1)-minigastrin for diagnostic application with (111)In for the in vivo detection of CCK-B receptor-expressing tissues. PMID- 12768331 TI - Improvement of signal-to-noise ratio using iterative reconstruction in a 99m Tc ECD split-dose injection protocol. AB - Split-dose injection using technetium-99m ethyl cysteinate dimer ((99m)Tc-ECD) and consecutive SPET measurements performed before and after acetazolamide (ACZ) loading was used to estimate the cerebral perfusion reserve. The disadvantage of the split-dose method is that the signal-to-noise ratio (S/N) of ACZ-loaded images is decreased by subtraction of the 1st SPET data (rest) from the 2nd SPET data (ACZ loaded). To improve the S/N of reconstructed images, we implemented an iterative reconstruction algorithm including the term of remaining radioactivity in the brain from the 1st injection. It was expected that this method (the "addition method") would improve the S/N of rest and ACZ images compared with the conventional subtraction method owing to exclusion of the subtraction process. To evaluate the effect of statistical noise, we estimated the percentage coefficient of variation (%COV) as a function of total photon counts (from 1.35 to 86.5 Mcounts/slice) by Monte Carlo simulation with equal-volume split-dose injection. The %COV of the 2nd SPET study was higher than that of the 1st (e.g. 50.3% for the 1st and 80.5% for the 2nd at a total count of 2.70 Mcounts/slice) when using the conventional subtraction method. By contrast, the %COV of the 1st and 2nd SPET studies was almost equivalent (e.g. 43.1% for the 1st and 41.4% for the 2nd at a total count of 2.70 Mcounts/slice) when using the addition method. We also determined the optimal injection dose ratio of the 2nd to the 1st SPET study, which provides the equivalent %COV value between the 1st and 2nd images. With the subtraction method, the optimal injection dose ratio of the 2nd to the 1st SPET study was approximately 2.0, while with the addition method it was approximately 1.0. The absolute value of %COV at the optimal injection dose was about 54% and 43% with the subtraction method and the addition method, respectively. The addition method gave a lower %COV value than the subtraction method even at the optimal injection dose ratio. In a clinical study, the addition method provided better quality images than the subtraction method. The ROI values of rest images estimated by the subtraction method were close to the results obtained with the addition method (ROI(sub)=1.01 ROI(add)-0.312, r=0.999). The ROI values of the ACZ images estimated by the subtraction method also agreed with the results obtained using the addition method, but the correlation was slightly worse (ROI(sub)=1.03 ROI(add)-2.23, r=0.995). Quantitative ROI values were quite similar between the methods. Our results demonstrated that the quality of reconstructed rest and ACZ-loaded images were significantly better with the addition method than with the conventional subtraction method. We conclude that the proposed method will be useful as a practical reconstruction algorithm to improve the S/N in an equal-volume split-dose injection protocol using (99m)Tc ECD. PMID- 12768332 TI - Stabilised 111In-labelled DTPA- and DOTA-conjugated neurotensin analogues for imaging and therapy of exocrine pancreatic cancer. AB - Neurotensin (NT) receptors are overexpressed in exocrine pancreatic cancer and Ewing's sarcoma. The potential utility of native NT in cancer diagnosis and therapy is, however, limited by its rapid degradation in vivo. Therefore, NT analogues were synthesised with modified lysine and arginine derivatives to enhance stability and coupled either to DTPA, to enable high specific activity labelling with indium-111 for imaging, or to DOTA, to enable high specific activity labelling with beta-emitting radionuclides, such as lutetium-177 and yttrium-90. Based on serum stability (4 h incubation at 37 degrees C in human serum) and receptor binding affinity, the five most promising analogues were selected and further evaluated in in vitro internalisation studies in human colorectal adenocarcinoma HT29 cells, which overexpress NT receptors. All five NT analogues bound with high affinity to NT receptors on human exocrine pancreatic tumour sections. The analogues could be labelled with (111)In to a high specific activity. The (111)In-labelled compounds were found to be very stable in serum. Incubation of HT29 cells with the (111)In-labelled analogues at 37 degrees C showed rapid receptor-mediated uptake and internalisation. The most promising analogue, peptide 2530 [DTPA-(Pip)Gly-Pro-(PipAm)Gly-Arg-Pro-Tyr-tBuGly-Leu-OH] was further tested in vivo in a biodistribution study using HT29 tumour-bearing nude mice. The results of this study showed low percentages of injected dose per gram tissue of this (111)In-labelled 2530 analogue in receptor-negative organs like blood, spleen, pancreas, liver, muscle and femur. Good uptake was found in the receptor-positive HT29 tumour and high uptake was present in the kidneys. Co injection of excess unlabelled NT significantly reduced tumour uptake, showing that tumour uptake is a receptor-mediated process. With their enhanced stability, maintained high receptor affinity and rapid receptor-mediated internalisation, the (111)In-labelled DTPA- and DOTA-conjugated NT analogues are excellent candidates for imaging and therapy of exocrine pancreatic cancer, peptide 2530 being the most promising analogue. PMID- 12768333 TI - 3-O-methyl-6-[18F]fluoro-L-DOPA and its evaluation in brain tumour imaging. AB - 3-O-Methyl-6-[(18)F]fluoro-L-DOPA (OMFD) is a major metabolite of 6-[(18)F]fluoro L-DOPA. Although synthesis of OFMD was primarily established to study the dopaminergic system, as it is an amino acid analogue, uptake in experimental tumours has been found. The aim of this study was to evaluate the applicability of OMFD for brain tumour imaging and to obtain initial estimates of whole-body biodistribution and radiation dosimetry in humans. Nineteen patients with suspected or confirmed brain tumours were investigated with OMFD and dynamic brain PET, complemented by whole-body PET in seven patients. Tracer kinetics were compared for normal brain and intracerebral lesions. Tissue accumulation was quantified with standardised uptake values (SUVs). Whole-body distribution in combination with tracer kinetics from animal experiments was used for the calculation of radiation dosimetry data. On the basis of OMFD PET, viable brain tumour was suspected in 16 patients with SUVs of 3.0+/-0.8 and a tumour to non tumour ratio of 1.9+/-0.5. Highest tumour and normal brain uptake occurred between 15 and 30 min, with a subsequent slow decrease. Late whole-body tracer distribution was uniform without specific organ accumulation. Elimination occurred via urine. The mean radiation dose to the whole body was estimated at 0.016 mSv/MBq, with the kidneys as dose-critical organ (0.033 mGy/MBq). In conclusion, OMFD enables the visualisation of brain tumours with SUVs similar to other fluorinated amino acids. The whole-body radiation exposure from OMFD is comparable to that from FDG imaging. PMID- 12768335 TI - Visualisation of the normal adrenals at SPET examination with 111In pentetreotide. AB - On single-photon emission tomography (SPET) examinations with indium-111 pentetreotide, we have often observed a small "hot" spot close to the upper portion of the left kidney. While it was initially believed that this represented a tumour in the tail of the pancreas or in the left adrenal, we now understand that it represents normal adrenal uptake. Since the adrenals have not been described as normally visualised, this uptake may interfere with the interpretation of the examination. We have studied how often the normal adrenals are visualised at such examinations. One hundred consecutive clinical (111)In pentetreotide examinations in adults including SPET of the abdomen and with normal findings were studied. All examinations with a hot spot attributable to the adrenals were, when available, compared with computed tomography (CT) or magnetic resonance imaging (MRI) of the abdomen performed on clinical grounds within 3 months before or at any time after the scintigraphy. In 11 patients both adrenals were visualised. In 27 patients only the left and in one only the right adrenal was visualised. CT or MRI examinations were available for comparison in 25 of these 39 patients. Two of these showed a pathological finding in the left adrenal. A conservative interpretation is that the normal adrenal is visualised on one or both sides in at least one-quarter of adults at SPET examination with (111)In-pentetreotide. PMID- 12768334 TI - 111In-DOTA- dPhe1-Tyr3-octreotide, 111In-DOTA-lanreotide and 67Ga citrate scintigraphy for visualisation of extranodal marginal zone B-cell lymphoma of the MALT type: a comparative study. AB - Somatostatin receptor (SSTR) scintigraphy and gallium-67 citrate ((67)Ga) scintigraphy have been used for visualisation of Hodgkin's lymphoma and non Hodgkin's lymphoma. However, experience with B-cell lymphoma of the mucosa associated lymphoid tissue (MALT) type is very limited. The aim of this study was to prospectively compare the (67)Ga scintigraphy results with those obtained by (111)In-DOTA- dPhe(1)-Tyr(3)-octreotide ((111)In-DOTA-TOCT) and (111)In-DOTA lanreotide ((111)In-DOTA-LAN) scintigraphy in patients with proven MALT-type lymphoma. Comparative scintigraphic examinations using (67)Ga, (111)In-DOTA-TOCT and (111)In-DOTA-LAN were performed in 18 patients (11 female and 7 male, median age 64+/-15 years) with histologically verified MALT-type lymphomas of various origin. Planar and single-photon emission tomography imaging acquisitions were performed after injection of a mean dose of 185+/-26 MBq (67)Ga and 165+/-20 MBq (111)In-DOTA-TOCT or (111)In-DOTA-LAN. All scintigraphic results were correlated with other conventional examinations including gastroscopy, colonoscopy, endosonoscopy, ophthalmologic investigation, CT of the thorax and abdomen and bone marrow biopsy. This comparative study showed that (67)Ga scintigraphy found abnormalities in 10 of 16 patients (63%) and detected 18 of 31 clinically involved sites (58%), but was false positive in three patients. (111)In-DOTA-TOCT found abnormalities in 9 of 15 patients (60%) and detected 15 of 27 clinical lesions (56%); it was false positive in two patients. (111)In-DOTA-LAN scintigraphy showed abnormalities in 7 of 11 patients (64%) and found 12 of 22 clinical lesions (55%). False-positive (111)In-DOTA-LAN scan results were found in two patients. For supra-diaphragmatic lesions, (67)Ga scintigraphy detected 12 of 16 sites (75%). (111)In-DOTA-TOCT scintigraphy revealed 7 of 15 lesions (47%). (111)In-DOTA-LAN showed 6 of 12 positive sites (50%). For infra-diaphragmatic involvement, the sensitivities of (67)Ga, (111)In-DOTA-TOCT and (111)In-DOTA-LAN were 40%, 67% and 60%, respectively. It is concluded that MALT-type lymphoma can be visualised by (67)Ga, (111)In-DOTA-TOCT and (111)In-DOTA-LAN scintigraphy. Although there were no statistically significant differences in patient-related and site-related sensitivities when using (67)Ga compared with (111)In-DOTA-TOCT and (111)In-DOTA-LAN, the sensitivity of (67)Ga tended to be superior to that of (111)In-DOTA-TOCT and (111)In-DOTA-LAN for supra-diaphragmatic lesions but inferior for infra-diaphragmatic involvement. In selected cases, the combination of (67)Ga and (111)In-DOTA-LAN or (111)In-DOTA-TOCT may increase the diagnostic efficiency in patients with MALT-type lymphoma. PMID- 12768336 TI - A hypothesis for the minimal overall structure of the mammalian plasma membrane redox system. AB - After a long period of frustration, many components of the mammalian plasma membrane redox system are now being identified at the molecular level. Some are apparently ubiquitous but are necessary only for a subset of electron donors or acceptors; some are present only in certain cell types; some appear to be associated with proton extrusion; some appear to be capable of superoxide production. The volume and variety of data now available have begun to allow the formulation of tentative models for the overall network of interactions of enzymes and substrates that together make up the plasma membrane redox system. Such a model is presented here. The structure discussed here is of the mammalian system, though parts of it may apply more or less accurately to fungal and plant cells too. Judging from the history of mitochondrial oxidative phosphorylation, it may be hoped that the development of models of the whole system - even if they undergo substantial revision thereafter - will markedly accelerate the pace of research in plasma membrane redox, by providing a coherent basis for the design of future experiments. PMID- 12768337 TI - Effect of dietary coenzyme Q and fatty acids on the antioxidant status of rat tissues. AB - Wistar rats were fed with different diets with or without supplement coenzyme Q(10) (CoQ(10)) and with oil of different sources (sunflower or virgin olive oil) for six or twelve months. Ubiquinone contents (CoQ(9) and CoQ(10)) were quantified in homogenates of livers and brains from rats fed with the four diets. In the brain, younger rats showed a 3-fold higher amount of ubiquinone than older ones for all diets. In the liver, however, CoQ(10) supplementation increased the amount of CoQ(9) and CoQ(10) in both total homogenates and plasma membranes. Rats fed with sunflower oil as fat source showed higher amounts of ubiquinone content than those fed with olive oil, in total liver homogenates, but the total ubiquinone content in plasma membranes was similar with both fat sources. Older rats showed a higher amount of ubiquinone after diets supplemented with CoQ(10). Two ubiquinone-dependent antioxidant enzyme activities were measured. NADH ferricyanide reductase activity in hepatocyte plasma membranes was unaltered by ubiquinone accumulation, but this activity increased slightly with age. Both cytosolic and membrane-bound dicumarol-sensitive NAD(P)H:(quinone acceptor) oxidoreductase (DT-diaphorase, EC 1.6.99.2) activities were decreased by diets supplemented with CoQ(10). Animals fed with olive oil presented lower DT diaphorase activity than those fed with sunflower oil, suggesting that the CoQ(10) antioxidant protection is strengthened by olive oil as fat source. PMID- 12768338 TI - A major integral protein of the plant plasma membrane binds flavin. AB - Abundant flavin binding sites have been found in membranes of plants and fungi. With flavin mononucleotide-agarose affinity columns, riboflavin-binding activity from microsomes of Cucurbita pepoL. hypocotyls was purified and identified as a specific PIP1-homologous protein of the aquaporin family. Sequences such as gi|2149955 in Phaseolus vulgaris, PIP1b of Arabidopsis thaliana, and NtAQP1 of tobacco are closely related. The identification as a riboflavin-binding protein was confirmed by binding tests with an extract of Escherichia coli cells expressing the tobacco NtAQP1 as well as leaves of transgenic tobacco plants that overexpress NtAQP1 or were inhibited in PIP1 expression by antisense constructs. When binding was assayed in the presence of dithionite, the reduced flavin formed a relatively stable association with the protein. Upon dilution under oxidizing conditions, the adduct was resolved, and free flavin reappeared with a half time of about 30 min. Such an association can also be induced photochemically, with oxidized flavin by blue light at 450 nm, in the presence of an electron donor. Several criteria, localization in the plasma membrane, high abundance, affinity to roseoflavin, and photochemistry, argue for a role of the riboflavin-binding protein PIP1 as a photoreceptor. PMID- 12768339 TI - Structure prediction for the di-heme cytochrome b561 protein family. AB - Atomic models possessing the common structural features identified for the cytochrome b(561) (cyt b(561)) protein family are presented. A detailed and extensive sequence analysis was performed in order to identify and characterize protein sequences in this family of transmembrane electron transport proteins. According to transmembrane helix predictions, all sequences contain 6 transmembrane helices of which 2-6 are located closely in the same regions of the 26 sequences in the alignment. A mammalian ( Homo sapiens) and a plant ( Arabidopsis thaliana) sequence were selected to build 3-dimensional structures at atomic detail using molecular modeling tools. The main structural constraints included the 2 pairs of heme-ligating His residues that are fully conserved in the family and the lipid-facing sides of the helices, which were also very well conserved. The current paper proposes 3-dimensional structures which to our knowledge are the first ones for any protein in the cyt b(561) family. The highly conserved His residues anchoring the two hemes on the cytoplasmic side and noncytoplasmic side of the membrane are in all proteins located in the transmembrane helices 2, 4 and 3, 5, respectively. Several highly conserved amino acids with aromatic side chain are identified between the two heme ligation sites. These residues may constitute a putative transmembrane electron transport pathway. The present study demonstrates that the structural features in the cyt b(561) family are well conserved at both the sequence and the protein level. The central 4-helix core represents a transmembrane electron transfer architecture that is highly conserved in eukaryotic species. PMID- 12768340 TI - Cytochrome b561 is not fatty acylated but acetylated at amino terminus in chromaffin vesicle membranes: an approach for the identification of posttranslational modification of transmembrane proteins. AB - We examined the nature of the posttranslational modification of bovine cytochrome b(561), a membrane-spanning protein and an essential component of neuroendocrine secretory vesicles. Matrix-assisted laser desorption and ionization time-of flight mass spectrometry (MALDI-TOF-MS) showed two populations in the partially digested fragments of cytochrome b(561), which were obtained by controlled treatment of cytochrome b(561)-proteoliposomes with trypsin. One population, containing the posttranslationally modified amino-terminal region, showed molecular masses which were by about 40 Da larger than the theoretical molecular masses. The other population, without the modified amino-terminal region, showed a reasonable matching with the theoretical masses. This result suggested that the posttranslational modification occurred only in the amino-terminal region. The amino-terminal peptide was isolated by tryptic peptide mapping followed by treatment with acylamino-acid-releasing enzyme. Amino acid sequence and MALDI-TOF MS analyses of the amino-terminal peptide showed that the initial Met residue was acetylated. There was no other posttranslational modification in the amino terminal region, such as covalent fatty acylation through an ester linkage to Ser or Thr residues. PMID- 12768341 TI - Partial purification and characterization of an ascorbate-reducible b-type cytochrome from the plasma membrane of Arabidopsis thaliana leaves. AB - The plant plasma membrane (PM) contains more than one b-type cytochrome. One of these proteins has a rather high redox potential (can be fully reduced by ascorbate) and is capable of transporting electrons through the PM. Four genes encoding proteins with considerable homology to the sequences of cytochrome b(561) proteins in the animal chromaffin granule membrane have recently been identified in the genome of Arabidopsis thaliana. In order to characterize the cytochrome b(561) located in the Arabidopsis PM, first PM vesicles were purified by aqueous polymer two-phase partitioning from the leaves of 9-week-old A. thaliana. PM proteins were solubilized by nonionic detergent, and the fully ascorbate-reducible b-type cytochrome was partially purified by anion-exchange chromatography steps. Potentiometric redox titration of the fraction, containing the fully ascorbate-reducible b-type cytochrome after the first anion-exchange chromatography step, revealed the presence of two hemes with redox potentials of 135 mV and 180 mV, respectively. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the fractions containing the fully ascorbate-reducible b-type cytochrome after the second anion-exchange chromatography step revealed the presence of a single polypeptide band at about 120 kDa. However, heat treatment (15 min, 90 degrees C) before electrophoresis was able to split the 120 kDa band into two bands with molecular masses of about 23 and 28 kDa. These values are lower than the apparent molecular mass for the fully ascorbate-reducible b-type cytochrome purified from Phaseolus vulgarishypocotyls (about 52 kDa) but are in good agreement with those characteristic for the cytochrome b(561) proteins purified from chromaffin granule membranes (about 28 kDa) and the four polypeptides predicted from the Arabidopsis genome (24-31 kDa). PMID- 12768342 TI - Differential distribution of ascorbic acid, peroxidase activity, and hydrogen peroxide along the root axis in Allium cepa L. and its possible relationship with cell growth and differentiation. AB - In this paper we show an asymmetrical distribution of apoplastic and symplastic ascorbic acid content, peroxidase activities and hydrogen peroxide along the root axis in Allium cepa L. For most of these metabolites, a marked gradient from the root apex to the onion base was observed and was different for apoplastic and symplastic compartments. In total homogenates, ascorbic acid content was higher in the zones closer to the apex and decreased towards the root base. However, an opposite pattern was observed in the apoplastic fraction. Peroxidase activities with guaiacol, ferulic acid, ascorbic acid, and coniferyl alcohol were also different depending on the evaluated zone and the fraction used (apoplastic or symplastic). In general, each activity had a specific and unique pattern. Immunodetection of peroxidase proteins in Western blots using anti-horseradish peroxidase and anti-ascorbate peroxidase antibodies revealed different bands at the different zones of the root. Hydrogen peroxide was detected by electron microscopy and was mainly found in cell walls of epidermis (or rhizodermis), meristem, and elongating cells. The number of cell walls showing hydrogen peroxide decreased dramatically towards the root base. The results suggest that the different zones of the root show specific requirements for ascorbic acid and hydrogen peroxide. Also, each fragment of the root seems to express specific peroxidase proteins. Different processes that take place at every part of the root, as cell proliferation and elongation near the root apex and gradual lignification and differentiation towards the root base are the key to explain the results. PMID- 12768343 TI - Salicylic acid changes the properties of extracellular peroxidase activity secreted from wounded wheat (Triticum aestivum L.) roots. AB - Wheat ( Triticum aestivum L.) roots released proteins showing peroxidase activity in the apoplastic solution in response to wound stress. Preincubation of excised roots with 1 mM salicylic acid at pH 7.0 enhanced the guaiacol peroxidase activity of the extracellular solution (so-called extracellular peroxidase). The soluble enzymes were partially purified by precipitation with ammonium sulfate followed by size exclusion and ion exchange chromatography. Despite an increase in the total activity of secreted peroxidase induced by pretreatment of excised roots with salicylic acid, the specific activity of the partially purified protein was significantly lower compared to that of the control. Purification of the corresponding proteins by ion exchange chromatography indicates that several isoforms of peroxidase occurred in both control and salicylic acid-treated samples. The activities of the extracellular peroxidases secreted by the salicylic acid-treated roots responded differently to calcium and lectins compared with those from untreated roots. Taken together, our data suggest that salicylic acid changes the isoforms of peroxidase secreted by wounded wheat roots. PMID- 12768344 TI - Superoxide synthase and dismutase activity of plasma membranes from maize roots. AB - Superoxide synthase and superoxide dismutase activity have been monitored in isolated maize ( Zea mays) root plasma membranes spectrophotometrically by determination of nitro-blue tetrazolium and cytochrome c reduction, respectively. Superoxide production was induced by NADH and NADPH, with similar kinetics and approaching saturation at 0.06 mM in the case of NADPH and 0.1 mM in the case of NADH, with rates of 18.6 +/- 5.0 and 21.8 +/- 7.2 nmol/min. mg of protein, respectively. These activities exhibited a broad pH optimum between pH 6.5 and 7.5. Diphenylene iodonium inhibited about 25% (10 microM DPI) and 40% (100 microM DPI) of this activity, imidazole inhibited about 20%, while KCN, a peroxidase inhibitor, did not show any significant inhibition. Superoxide-dismutating activity was shown to occur in the same isolates and depended on the quantity of plasma membrane protein present. Growth of plants on salicylic acid prior to membrane isolation induced a rise in the activity of both of the enzymes by 20 35%, suggesting their coordinated action. PMID- 12768345 TI - Redox-related peroxidative responses evoked by methyl-jasmonate in axenically cultured aeroponic sunflower (Helianthus annuus L.) seedling roots. AB - Methyl-jasmonate (MeJA) has been proposed to be involved in the evocation of defense reactions, as the oxidative burst in plants, substituting the elicitors or enhancing their effect. 48 h dark- and sterilely cultured (axenic) aeroponic sunflower seedling roots excised and treated with different concentrations of MeJA showed a strong and quick depression of the H(+) efflux rate, 1.80 microM MeJA totally stopping it for approximately 90 min and then reinitiating it again at a lower rate than controls. These results were wholly similar to those obtained with nonsterilely cultured roots and have been interpreted as mainly based on H(+) consumption for O(2)(*-) dismutation to H(2)O(2). Also K(+) influx was strongly depressed by MeJA, even transitorily reverting to K(+) efflux. These results were consistent with those associated to the oxidative burst in plants. MeJA induced massive H(2)O(2) accumulation in the middle lamella and intercellular spaces of both the root cap cells and the inside tissues of the roots. The native acidic extracellular peroxidase activity of the intact (nonexcised) seedling roots showed a sudden enhancement (by about 52%) after 5 min of MeJA addition, maintained for approximately 15 min and then decaying again to control rates. O(2) uptake by roots gave similar results. These and other results for additions of H(2)O(2) or horseradish peroxidase, diphenylene iodonium, and sodium diethyldithiocarbamate trihydrate to the reaction mixture with roots were all consistent with the hypothesis that MeJA induced an oxidative burst, with the generation of H(2)O(2) being necessary for peroxidase activity. Results with peroxidase activity of the apoplastic fluid were in accordance with those of the whole root. Finally, MeJA enhanced NADH oxidation and inhibited hexacyanoferrate(III) reduction by axenic roots, and diphenylene iodonium cancelled out these effects. Redox activities by CN(-)- preincubated roots were also studied. All these results are consistent with the hypothesis that MeJA enhanced the NAD(P)H oxidase of a redox chain linked to the oxidative burst, so enhancing the generation of O(2)(*-) and H(2)O(2), O(2) uptake, and peroxidase activity by roots. PMID- 12768346 TI - Copper-mediated oxidative burst in Nicotiana tabacum L. cv. Bright Yellow 2 cell suspension cultures. AB - In cell suspension cultures of Nicotiana tabacum L. cv. Bright Yellow 2 (BY-2) a rapid and concentration-dependent accumulation of H(2)O(2) is induced by excess concentrations of copper (up to 100 microM). This specific and early response towards copper stress was shown to be extracellular. Addition of 300 U of catalase per ml decreased the level of H(2)O(2). Superoxide dismutase (5 U/ml) induced an increase in H(2)O(2) production by 22.2%. This indicates that at least part of the H(2)O(2) is produced by dismutation of superoxide. Pretreatment of the cell cultures with the NAD(P)H oxidase inhibitors diphenylene iodonium (2 and 10 microM) and quinacrine (1 and 5 mM) prevented the generation of H(2)O(2) under copper stress for 90%. The influence of the pH on the H(2)O(2) production revealed the possible involvement of cell-wall-dependent peroxidases in the generation of reactive oxygen species after copper stress. PMID- 12768347 TI - Proton conduction through full-length gp91phox requires histidine 115. AB - The NADPH oxidase of neutrophils is a transmembrane electron transfer complex, containing a flavin adenine dinucleotide and two hemes, all of which are suggested to be contained within gp91 (phox), one of four subunits of the enzyme. The transfer of electrons through the NADPH oxidase is associated with an efflux of protons. gp91 (phox) has previously been demonstrated to function as the proton conduction pathway. The mutation of histidines 111, 115, and 119 to leucines and of histidine 115 to leucine within the N-terminal 230-amino-acid fragment of gp91 (phox) has previously been demonstrated to result in the loss of proton conduction through this N-terminal fragment. In this paper we have investigated the role of these histidines in proton conduction by the full-length gp91 (phox). Stable CHO cell lines were established which expressed full-length gp91 (phox) in which histidines 111, 115, and 119 had been mutated to leucines (CHO91H111/115/119) and in which histidine 115 had been mutated to leucine (CHO91H115L). The expression of gp91 (phox) and its cellular localisation in these cell lines were comparable between wild-type and the mutant gp91 (phox). The mutation of histidines 111, 115, and 119 to leucines or just histidine 115 to leucine resulted in an almost total loss of both the arachidonate-activated influx and efflux of protons, in comparison with that observed for wild-type gp91 (phox). Therefore, histidine 115 is required for proton conduction by both full length gp91 (phox) and the N-terminal 230-amino-acid fragment of gp91 (phox). Histidine 115 has recently been proposed to act as a coordinating ligand for the outer heme iron of the NADPH oxidase. On the basis of observations for cytochrome c oxidase, we propose a model for this dual role of histidine 115. PMID- 12768348 TI - Inhibition of neutral Mg2+-dependent sphingomyelinase by ubiquinol-mediated plasma membrane electron transport. AB - Sphingomyelin is an abundant constituent of the plasma membranes of mammalian cells. Ceramide, its primary catabolic intermediate, has emerged as an important lipid signaling molecule. Previous work carried out by our group has documented that plasma membrane Mg(2+)-dependent neutral sphingomyelinase can be effectively inhibited by exogenous ubiquinol. In this work, we have tested whether or not plasma-membrane-associated electron transport can also achieve this inhibition through endogenous ubiquinol. Our results have shown that Mg(2+)-dependent neutral sphingomyelinase in isolated plasma membranes was inhibited by NAD(P)H under conditions where ubiquinone is reduced to ubiquinol. This inhibition was potentiated in the presence of an extra amount of NAD(P)H:(quinone acceptor) oxidoreductase 1 (EC 1.6.99.2). Depletion of plasma membranes from lipophilic antioxidants by solvent extraction abolished the inhibition by reduced pyridine nucleotides without affecting the sensitivity of the neutral sphingomyelinase to exogenous ubiquinol. Reconstitution of plasma membranes with ubiquinone restored the ability of NAD(P)H to inhibit the enzyme. Our results support that the reduction of endogenous ubiquinone to ubiquinol by NAD(P)H-driven electron transport may regulate the activity of the plasma membrane neutral sphingomyelinase. PMID- 12768349 TI - Redox signaling of NF-kappaB by membrane NAD(P)H oxidases in normal and malignant cells. AB - Evidence is rapidly accumulating that low-activity NAD(P)H oxidases homologous to that in phagocytic cells generate reactive oxygen species as signaling intermediates. In this review we discuss evidence that signaling NAD(P)H oxidases in part influence normal and malignant cell division by activating the redox regulated transcription factor nuclear factor kappaB. The roles of growth regulatory NAD(P)H oxidases in human airway smooth muscle and malignant melanoma are used as examples. PMID- 12768350 TI - Regeneration of lipophilic antioxidants by NAD(P)H:quinone oxidoreductase 1. AB - The aim of this work was to study the activity of NAD(P)H:(quinone acceptor) oxidoreductase 1 (EC 1.6.99.2) in the regeneration of lipophilic antioxidants, alpha-tocopherol, and reduced-coenzyme Q analogs. First, we tested whether or not two isoforms of the NAD(P)H:(quinone acceptor) oxidoreductase 1 designated as "hydrophilic" and "hydrophobic" (H. J. Prochaska and P. Talalay, Journal of Biological Chemistry 261: 1372-1378, 1986) show differential enzyme activities towards hydrophilic or hydrophobic ubiquinone homologs. By chromatography on phenyl Sepharose, we purified the two isoforms from pig liver cytosol and measured their reduction of several ubiquinone homologs of different side chain length. We also studied by electron paramagnetic resonance the effect of NAD(P)H:(quinone acceptor) oxidoreductase 1 on steady-state levels of chromanoxyl radicals generated by linoleic acid and lipooxygenase and confirmed the enzyme's ability to protect alpha-tocopherol against oxidation induced with H(2)O(2) Fe(2+). Our results demonstrated that the different hydrophobicities of the isoforms do not reflect different reactivities towards ubiquinones of different side chain length. In addition, electron paramagnetic resonance studies showed that in systems containing the reductase plus NADH, levels of chromanoxyl radicals were dramatically reduced. Morever, in the presence of oxidants, alpha tocopherol was preserved by NAD(P)H:(quinone acceptor) oxidoreductase 1, supporting our hypothesis that regeneration of alpha-tocopherol may be one of the physiologic functions of this enzyme. PMID- 12768351 TI - Role of reactive oxygen species in the response of barley to necrotrophic pathogens. AB - The interactions between Hordeum vulgare(barley) and two fungal necrotrophs, Rhynchosporium secalis and Pyrenophora teres (causal agents of barley leaf scald and net blotch), were investigated in a detached-leaf system. An early oxidative burst specific to epidermal cells was observed in both the susceptible and resistant responses to R. secalis, and later on, a second susceptible-specific burst was observed. Time points of the first and the second burst correlated closely with pathogen contact to the plasma membrane and subsequent cell death, respectively. HO(2)(*)/O(2)(-) levels in resistant and susceptible responses to P. teres were limited in comparison. During later stages, HO(2)(*)/O(2)(-) was only detected in 2 to 3 epidermal cells immediately adjacent to phenolic browning and cell death observed during the susceptible response. However, H(2)O(2) was detected in the majority of mesophyll cells adjacent to the observed lesion caused by P. teres. In contrast to observations during challenge with R. secalis, no direct contact between P. teres and the plasma membrane at sites of reactive oxygen species production was evident. Preinfiltration of leaves with antioxidants prior to challenge with either pathogen had no effect on resistance responses but did limit the growth of the pathogens and inhibit the extent of cell death during susceptible responses. These results suggest a possible role for reactive oxygen species in the induction of cell death during the challenge of a susceptible plant cell with a necrotrophic fungal leaf pathogen. PMID- 12768352 TI - Biochemical entities involved in reactive oxygen species generation by human spermatozoa. AB - Spermatozoa were the first cell type suggested to generate reactive oxygen species. However, a lack of standardization in sperm preparation techniques and the obfuscating impact of contaminating cell types in human ejaculates have made it difficult to confirm that mammalian germ cells do, in fact, make such reactive metabolites. By identifying, on a molecular level, those entities involved in reactive oxygen species generation and demonstrating their presence in spermatozoa, the role of redox chemistry in the control of sperm function can be elucidated. Two major proteins have apparently been identified in this context, namely, NOX5, a calcium-activated NADPH oxidase, and nitric oxide synthase. Understanding the involvement of these enzymes in sperm physiology is essential if we are to understand the causes of oxidative stress in the male germ line. PMID- 12768353 TI - Reduction of potassium tellurite to elemental tellurium and its effect on the plasma membrane redox components of the facultative phototroph Rhodobacter capsulatus. AB - Anaerobically light-grown cells of Rhodobacter capsulatus B100 are highly resistant to the toxic oxyanion tellurite (TeO(3)(2-); minimal inhibitory concentration, 250 microg/ml). This study examines, for the first time, some structural and biochemical features of cells and plasma membrane fragments of this facultative phototroph grown in the presence of 50 microg of K(2)TeO(3) per ml. Through the use of transmission microscopy and X-ray microanalysis we show that several "needlelike" shaped granules of elemental tellurium are accumulated into the cytosol near the intracytoplasmic membrane system. Flash-spectroscopy, oxygen consumption measurements, and difference spectra analysis indicated that membrane vesicles (chromatophores) isolated from tellurite-grown cells are able to catalyze both photosynthetic and respiratory electron transport activities, although they are characterized by a low c-type cytochrome content (mostly soluble cytochrome c(2)). This feature is paralleled by a low cytochrome c oxidase activity and with an NADH-dependent respiration which is catalyzed by a pathway leading to a quinol oxidase (Qox) inhibited by high (millimolar) concentrations of cyanide (CN(-)). Conversely, membranes from R. capsulatus B100 cells grown in the absence of tellurite are characterized by a branched respiratory chain in which the cytochrome c oxidase pathway (blocked by CN(-) in the micromolar range) accounts for 35-40% of the total NADH-dependent oxygen consumption, while the remaining activity is catalyzed by the quinol oxidase pathway. These data have been interpreted to show that tellurite resistance of R. capsulatus B100 is characterized by the presence of a modified plasma-membrane associated electron transport system. PMID- 12768354 TI - Effects of 5-HT(6) receptor blockade on the neurochemical outcome of antidepressant treatment in the frontal cortex of the rat. AB - Using in vivo microdialysis in the freely moving rat we have examined the effects of 5-HT(6) receptor antagonism on the neurochemical outcome of antidepressant treatment. Acute administration of both desipramine (10 mg/kg s.c.) and venlafaxine (10 mg/kg s.c.) produced a 2 fold increase in extracellular noradrenaline (NA) but no change in frontal cortex dopamine (DaA), 5-HT or glutamate. Fluoxetine (20 mg/kg s.c.) produced no change in extracellular levels of any of the neurotransmitters examined. SB-271046 produced a 3 fold increase in extracellular glutamate. Combination treatment of SB-271046 with each antidepressant produced no change in the antidepressant-induced changes in NA, DA or 5-HT. In contrast, both fluoxetine and venlafaxine attenuated the SB-271046 induced increase in extracellular glutamate, suggesting that 5-HT and possibly NA may be having an inhibitory action on the excitatory pathways enhanced by 5-HT(6) receptor blockade. Furthermore, these data indicate that the neurochemical effects induced by NA and/or 5-HT reuptake inhibitors are not enhanced by 5-HT(6) receptor blockade indicating that 5-HT(6) receptor antagonists are unlikely to augment the therapeutic efficacy of these types of antidepressants. PMID- 12768355 TI - Extrastriatal dopamine D(2) receptors in Parkinson's disease: a longitudinal study. AB - Most antiparkinsonian drugs are known to act through central dopamine D(2) receptor agonism. A previous longitudinal positron emission tomography (PET) study has indicated that, in the striatum of Parkinson's disease (PD) patients, dopamine D(2) receptor binding declines at a relatively fast annual rate of 2-4% (compared to the rate of <1%/year in healthy individuals). In the present study, the examination of longitudinal changes in D(2) receptors was extended to extrastriatal brain regions in PD. Eight early PD patients were examined twice with PET, approximately 3 years apart, using a high-affinity extrastriatal D(2)/D(3) receptor tracer, [(11)C]FLB 457. Both the MRI-referenced region-of interest method and the voxel-based statistical analysis method were used independently in the analysis. Regional D(2)-like availabilities (binding potentials) in the left dorsolateral prefrontal cortex, the left temporal cortex and the left and right medial thalami were significantly decreased at the second examination by 20-37% (corresponding to an annual decline of 6-11%). Thus, the annual loss of extrastriatal D(2) availability in PD is up to three times faster than the rate previously reported in the putamen. Our longitudinal study shows first evidence concerning cortical D(2) receptor loss in the progression of PD, although it is not possible to distinguish between the effects of the therapy and the disease. PMID- 12768356 TI - Repeated rating improves value of diagnostic dopaminergic challenge tests in Parkinson's disease. AB - Clinicians use acute challenges with levodopa (LD) and/or apomorphine (A) for diagnostic dopaminergic response tests in Parkinson's disease (PD) patients. We consecutively compared the value of both drugs with performance of repeated ratings and adverse effect recording. Oral administration of 200 mg LD was superior to subcutaneous injection of 4 mg A in terms of tolerability and onset of temporary UPDRS motor score decline ([previously untreated PD patients] LD: 4.02 [mean] +/- 2.45 [SD] [significant decrease: p = 1.42 E-07] vs. A: 1.58 +/- 3.38 [not significant decrease: p = 0.14], p = 0.0009; [treated PD patients] LD: 7.71 +/- 4.35 [significant decrease: p = 2.48 E-06] vs. A: 5.19 +/- 4.32 [significant decrease: p = 7.83 E-05], p = 0.07). We suggest diagnostic acute challenge test performance with LD as first- and A as second choice due to better tolerability and valuation in combination with repeated scoring procedures to improve sensitivity and specifity. PMID- 12768357 TI - Acute double-blind, placebo-controlled sleep laboratory and clinical follow-up studies with a combination treatment of rr-L-dopa and sr-L-dopa in restless legs syndrome. AB - In a double-blind, placebo-controlled randomized crossover trial, the acute efficacy of a combination treatment of 100 mg regular-release (rr) and 100 mg sustained-release (sr) L-dopa/benserazide in RLS was investigated by means of sleep laboratory methods, with a subsequent open clinical follow-up for 4 weeks. 21 RLS patients classified according to ICSD and IRLSSG criteria were included; 18 completed the study. Objective sleep quality was determined by polysomnography (PSG) in 3 subsequent nights (adaptation/screening, placebo and drug night), subjective sleep and awakening quality was evaluated by rating scales, objective awakening quality by psychometric tests. Clinical follow-up consisted of daily ratings of subjective sleep and awakening quality (SSA) and VAS for RLS symptomatology ratings, completion of the RLS (IRLSSG) Scale weekly and the Zung Depression (SDS) and Anxiety (SAS) Scale, Quality of Life Index, Pittsburgh Sleep Quality Index and Epworth Sleepiness Scale before and after therapy. Acute L dopa/benserazide significantly (p < 0.001) and markedly (75%) decreased the target variable PLM/h of sleep as well as all other RLS/PLM variables, but failed to improve objective sleep efficiency and subjective sleep quality in comparison to placebo. After 4 weeks of therapy, however, subjective sleep and awakening quality also improved significantly. While RLS/PLM measures showed an immediate significant and marked response to the combination therapy subjective sleep quality only improved after chronic treatment. PMID- 12768358 TI - Nerve growth factor response to excitotoxic lesion of the cholinergic basal forebrain is slightly impaired in aged rats. AB - Nerve growth factor (NGF) promotes survival and function of basal forebrain cholinergic neurons. We studied NGF and choline acetyltransferase (ChAT) activity after partial quisqualic acid induced lesions of the basal forebrain in 3 and 27 months-old rats, in order to investigate whether NGF-related regeneration is disturbed in old age. 2 weeks post lesion, ChAT activity decreased by 25 to 32% in adult and old rats. 3 months post lesion, the ChAT deficit receded in adult rats, but remained unchanged in old rats. 2 weeks post lesion, NGF levels were reduced by 36 to 44%, but there was no significant difference between adult and old rats. 3 months post lesion, we found increased NGF levels by 44% in the posterior cortex of adult rats. These results indicate that the compensatory NGF increase in the posterior cortex after partial cholinergic lesion of the basal forebrain is slightly impaired in old age. PMID- 12768359 TI - Brain region-dependent increases in beta-amyloid and apolipoprotein E levels in hypercholesterolemic rabbits. AB - Recent studies indicate a possible link between serum cholesterol level, beta amyloid (Abeta) peptide concentrations, and the incidence of Alzheimer's disease (AD). In the present report, the effects of dietary cholesterol on Abeta and apolipoprotein E (APOE) levels in several brain regions were examined using diet induced hypercholesterolemic rabbits as the animal model. Increased dietary cholesterol levels increased Abeta concentrations in temporal cortex (p = 0.02). A similar trend was observed in the frontal cortex (p = 0.06), yet not in the cerebellum. Interestingly, the regional levels of Abeta in the hypercholesterolemic rabbit paralleled the amyloid pathology observed in AD brain. Elevated APOE levels were also noticed in temporal (p < 0.01) and frontal (p < 0.01) cortices, but not in cerebellum, in the rabbit fed with cholesterol abundant diet. These results suggest that high serum cholesterol levels could induce the elevation of brain APOE, which may play a role in aggravating the Abeta accumulation. PMID- 12768360 TI - Possible increased risk for Alzheimer's disease associated with neprilysin gene. AB - Neprilysin has recently been reported to be the major physiological Abeta degradating enzyme. In this study we describe a new biallelic polymorphism in the 3'UTR of the neprilysin gene in a representative population sample. The (*)159C/C genotype was found to be associated with an increased risk for Alzheimer's disease in an age-dependent manner. Adjusting for sex and APOE status, an odds ratio of 2.74 (p < 0.05) was observed among patients under 75 years old. PMID- 12768361 TI - The clinical use of P300 event related potentials for the evaluation of cholinesterase inhibitors treatment in demented patients. AB - Centrally acting cholinesterase inhibitors (ChEIs) improve cognitive functions in Alzheimer's disease (AD) and other forms of dementia. Evaluation of treatment efficacy is based mainly on subjective assessment methods such as standardized neuropsychological tests. Therefore, an additional objective tool for the evaluation of drug response would be most helpful.Thirty-two patients suffering from dementia of several etiologies were treated with ChEIs (tacrine 19, donepezil 5, rivastigmine 8). Cognitive response was assessed pre ChEIs initiation (baseline) and after 26 weeks, as optimal tolerated doses were achieved and maintained (endpoint). Evaluation included repeated measurements of Mini Mental State Examination (MMSE), Alzheimer's disease assessment scale cognitive part (ADAS-cog) and P300. For statistical analysis we used ANOVA with repeated measures and Pearson correlation coefficient. Results demonstrated improvement of mean ADAS-cog by 2.0 points (from 29.4, n = 31 to 27.4, n = 29; p = 0.08) while MMSE remained almost unchanged (20.1, n = 29 to 19.8, n = 28). Mean P300 latency reduced significantly by 24 ms (from 383 +/- 7.9 msec, n = 32 to 359 +/- 7 msec, n = 32; p = 0.0001). However mean amplitudes did not change significantly from baseline to endpoint (13.5 +/- 6.2, n = 31 to 12.8 +/- 6.1, n = 31). Significant correlations were found between mean ADAS-cog and mean P300 latency at baseline and end-point (R = 0.485 p = 0.019, R = 0.626 p = 0.001 respectively, n = 23) and between mean MMSE and P300 latency at baseline and endpoint (R = -0.420 p = 0.046, R = -0.703 p < 0.001 respectively, n = 23). Our data suggests that P300 is a reliable instrument for assessment of cognitive response to ChEIs in demented patients. PMID- 12768363 TI - Rett Syndrome -- an update. AB - Rett syndrome is a progressive, usually sporadic and rarely familial, disabling neurodevelopmental disorder with onset in early childhood presenting clinically with mental retardation, behavioral changes, late movement disturbances, loss of speech and hand skills, ataxia, apraxia, irregular breathing with hyperventilation while awake, and frequent seizures. It occurs almost exclusively in females with an estimated prevalence of 1 in 10-22000 births and is considered a manifestation of defective brain maturation caused by dominant mutation of the MeCP2 gene encoding the transcriptional repressor methyl-CpG-binding protein 2 related to the Xq28 locus. Although many different mutations of this protein are being studied in humans and in mice, the molecular pathogenesis of this disorder remains unclear. Electroencephalography is abnormal in the final stages of the syndrome. Neuroimaging showing brain atrophy may be required for differential diagnosis that includes neurodegenerative and metabolic disorders. Neuropathology shows decreased brain growth and reduced size of individual neurons, with thinned dendrites in some cortical layers and abnormalities in substantia nigra (decreased neuromelanin content), suggestive of deficient synaptogenic development, probably starting before birth. Neurometabolic changes include reduced levels of dopamine, serotonin, noradrenalin, choline acetyltransferase (ChAT), nerve growth factors, endorphines, glutamate, and other amino acids and their receptor levels in brain. Current treatment includes symptomatic, anticonvulsive and physiotherapy. PMID- 12768362 TI - Reduced phosphorylation of cyclic AMP-responsive element binding protein in the postmortem orbitofrontal cortex of patients with major depressive disorder. AB - In this study, we examined the amounts of cAMP-responsive element binding protein (CREB) and its phosphorylated form in homogenate preparations from postmortem orbitofrontal cortices of antidepressant drug-free patients with major depressive disorder and age-matched controls by immunoblotting. Immunoreactivies of both CREB and phosphorylated CREB were significantly decreased in depressive subjects compared to controls. The immunoreactivity of phosphorylated CREB was diminished to a greater extent than that of CREB in depressive patients. It has been indicated from animal studies that a transcription factor likely mediates neural plasticity in the mammalian brain and neural tissues. Our results suggest that alterations in the cAMP signaling system, especially in CREB, may be involved in the pathophysiology of depression and be potential targets for antidepressant treatment. PMID- 12768364 TI - Clinicopathologic and technical factors associated with the uptake of radiocolloid by sentinel nodes in patients with breast cancer. AB - PURPOSE: Sentinel-node biopsy is becoming widely accepted in breast cancer treatment. Using the radioisotope technique, a lower risk of identification failure is related to the amount of radiocolloid in the sentinel nodes. The aim of this study was to identify the factors associated with the colloidal uptake of the sentinel nodes. METHODS: Technetium-labeled colloid was injected peritumorally, with or without subdermal injection. According to the maximum radioactivity of the sentinel nodes, patients were divided into high (>/=100 counts/s) or low (<100 counts/s) uptake groups. The uptake was compared in relation to the clinicopathologic and technical features. RESULTS: The sentinel node was identified in 183 of 186 patients (98.4%), with 60 and 123 patients in the low- and high-uptake groups (mean: 39 and 1003 counts/s), respectively. Multivariate analysis showed that an age of 65 years or older and a sentinel-node size of 8 mm or more were significantly more predominant in the low-uptake group. CONCLUSION: Care must be taken when performing sentinel-node biopsy, especially for aged patients and for those with large sentinel nodes. The optimal technique should be determined on the basis of these results. PMID- 12768365 TI - Intratumoral heterogeneity of genetic changes in primary colorectal carcinomas with metastasis. AB - PURPOSE: To elucidate the intratumoral distribution of genetic changes, surgically resected colorectal carcinomas were investigated regarding their loss of heterozygosity in the regions of tumor suppressor genes and the mutation of ras genes. METHODS: Full-thickness fresh tumor slices from 23 colorectal carcinomas were removed and divided into multiple specimens, which were then submitted separately for DNA and histopathological analyses. The loss of heterozygosity was analyzed in 23 primary carcinomas and 9 metastasized carcinomas by the use of restriction fragment-length polymorphism markers on chromosome 1p, 5q, 17p, 18q, and 22q. RESULTS: Intratumoral heterogeneity was identified in 11 of 23 primary carcinomas (47.8%) and we could successfully map the regional genetic variation. In both stages I and II, 1 of 5 cases (20%); in stage III, 3 of 6 cases (50%); and in stage IV, 6 of 7 cases (85.7%) were heterogeneous. With respect to venous invasion positive primary carcinomas, hepatic metastasis occurred in 75% (6/8) of the heterogeneous carcinomas, whereas hepatic metastasis occurred in only 12.5% (1/8) of homogeneous carcinomas. CONCLUSION: The present results suggest that the existence of intratumoral heterogeneity, which may reflect genetic instability, may thus play a role in the enhancement of aggressive progression and the metastasis of colorectal carcimomas. PMID- 12768366 TI - Minilaparotomy approach to colon cancer. AB - PURPOSE: Many studies have proved the feasibility and safety of a laparoscopic colectomy in comparison to a conventional laparotomy. However, a laparoscopic colectomy requires a minilaparotomy incision to perform the operative procedure. We have introduced a minilaparotomy technique which can perform all the operative procedures through incisions measuring from 3 to 7 cm in length. METHODS: A retrospective comparison of the outcome after a potentially curative resection of colon cancers via minilaparotomy (July 2000 to May 2002) and by conventional laparotomy (May 1997 to June 2000) is reported. RESULTS: The patient cohort consisted of 27 minilaparotomy cases and 24 conventional laparotomy cases. The patients' characteristics were similar in the two groups. The oncological clearance, in terms of the length of resected specimens, was similar in the two groups, whereas the number of lymph nodes removed was significantly higher in the minilaparotomy group. In addition, the mean operation time, blood loss, length of the laparotomy incision, postoperative time to walking, starting oral intake, and postoperative hospitalization were significantly smaller in the minilaparotomy group. CONCLUSION: Our minilaparotomy approach maintained the same curative resection for colon cancers as a conventional laparotomy, but it was less invasive and allowed for an earlier recovery and hospital discharge than conventional laparotomy. The minilaparotomy approach is thus considered to be an attractive alternative to conventional colon surgery. PMID- 12768367 TI - The influence of viral genotypes and rejection episodes on the recurrence of hepatitis C after liver transplantation. AB - PURPOSE: The aim of this study was to report the influence of hepatitis C virus (HCV) genotype and rejection episodes on the outcome of orthotopic liver transplantation (OLT), hepatitis recurrence, and progression to graft cirrhosis after OLT. METHODS: Fifty-three patients who all had undergone OLT for end-stage liver cirrhosis were selected for this study. Hepatitis C genotype was determined. Recurrent hepatitis and rejection were diagnosed based on elevated liver function tests and a liver biopsy. RESULTS: The patients were followed up for a mean of 51.9 +/- 34.3 months. The cumulative survival rate was no different in OLT for hepatitis C and OLT for all other liver diseases. After OLT, serum HCV RNA was detected in 93%. Histological recurrence occurred in 85% of all patients. The 1-, 3-, and 5-year recurrence rates were 48%, 77%, and 85%, respectively. Of the 41 patients with recurrent hepatitis C, 4 (10%) had cirrhosis, 18 (44%) had hepatitis with fibrosis, and 91 (46%) had hepatitis without fibrosis at the end of follow-up. A total of 32% of the patients were infected by HCV genotype 1b and 68% by other HCV genotypes. The recurrence rates were significantly higher in patients infected with genotype 1b than in those with other genotypes ( p = 0.04). Twenty of 48 patients (42%) experienced acute rejection. There was a strong association between the number of rejection episodes and the incidence of HCV-related cirrhosis ( p < 0.01). CONCLUSION: Our findings showed the genotype 1b to result in a higher recurrence rate after OLT. On the other hand, rejection episodes were associated with a more rapid progression to graft cirrhosis. PMID- 12768368 TI - Doxycycline treatment in a model of early abdominal aortic aneurysm. AB - PURPOSE: To evaluate the effects doxycycline (Dox) in animal models of early abdominal aortic aneurysm. METHODS: Of 43 male Wistar rats, 33 underwent intraluminal perfusion of the abdominal aorta with thioglycolate plus plasmin to reproduce early aortic aneurysm. These rats then were treated for 7 days with subcutaneous injections of Dox or saline. The 10 remaining rats underwent intra aortic perfusion with saline and were injected subcutaneously with saline. On day 7, the rats were killed after abdominal aortic diameters were measured. Some aortic specimens were examined microscopically after elastica van Gieson (EVG) and hematoxylin-eosin (H&E) staining. In other specimens, the matrix metalloproteinase (MMP) activity in tissue extracts was evaluated by gelatin zymography. RESULTS: Among the thioglycolate plus plasmin-perfused rats, the degree of aortic dilation was less in Dox-treated than in saline-treated rats. EVG staining indicated that Dox maintained a nearly normal pattern of elastic lamellae and normal medial elastin thickness. The aortic inflammatory response was not suppressed by Dox in H&E staining. In gelatin zymography, Dox reduced the MMP-9 activity, but did not significantly change either MMP-2 or the percentage of activated MMP-2. CONCLUSIONS: Dox inhibited experimental aneurysmal dilation by preserving medial elastin. This effect involved the suppression of MMP-9 but not of the MMP-2 activity. PMID- 12768369 TI - Identification of differentially expressed mRNAs of the autologous vein graft after an arterial bypass in mongrel dogs. AB - PURPOSE: Intimal hyperplasia (IH) is characterized by vascular smooth muscle cell (VSMC) proliferation in the intima and subsequent accumulation of extracellular matrix. A variety of factors that might be considered as possible VSMC mitogens induce the specific gene expression of VSMC. This study was designed to identify differentially expressed mRNA by method using an mRNA differential display. METHODS: A bilateral femoropopliteal reverse saphenous vein bypass was performed on both hind limbs of mongrel dogs. At 4 and 8 weeks after the implantation, the vein bypass grafts were harvested. The total RNAs were extracted from each specimen and transcribed into the cDNAs. Amplified cDNAs using 16 sets of primers were separated on DNA sequencing gel. Differentially displayed bands were excised from the gel, cloned, and then sequenced. The sequences were compared with the National Center of Biotechnology Information nonredundant sequence database using the Basic Local Alignment Search Tool (BLAST) program. RESULTS: Forty-three bands were cloned and sequenced. A computer search revealed that 11 of them had similarities to known genes while 32 cDNAs had no similarities to any registered DNA sequence. CONCLUSIONS: We found several genes to be related to IH. Some of them had already had their function and sequence identified whereas some of them have yet to be identified. Further studies are necessary to determine the precise relationship between these genes and IH. PMID- 12768370 TI - Coexisting aortocolic and aortovesical fistulae in an abdominal aortic aneurysm: report of a case. AB - Aortoenteric fistula is a rare complication in abdominal aortic aneurysms (AAA), and an aortovesical fistula is even more rare. This is a report of a case of aortocolic and aortovesical fistulae in a 64-year-old man with AAA who was treated with an aortobiiliac bypass, a primary repair of the fistulated bladder, and a sigmoid colostomy. PMID- 12768371 TI - Basaloid-squamous carcinoma of the esophagus treated by preoperative chemotherapy: report of two cases. AB - Basaloid squamous carcinoma (BSC) of the esophagus has been associated with a poor outcome after surgery. We herein report two patients with esophageal BSC treated by preoperative chemotherapy. Patient 1 was a 55-year-old man who presented with a tumor of the middle esophagus diagnosed as BSC. He was treated by chemotherapy using a combination of 5-fluorouracil (5-FU: 750 mg/m(2), 1st-5th day, 24-h continuous infusion) and cisplatin (CDDP: 75 mg/m(2), 1st day, drip infusion per 2 h) before surgery, because of lymph node metastases of the mediastinum and around the left gastric artery. Even though the metastatic nodes were reduced and an esophagectomy was performed, the patient died of recurrence 12 months after chemotherapy. Patient 2 was a 57-year-old man who demonstrated BSC of the esophagus with direct invasion to the discending aorta, who was treated by preoperative chemotherapy using the same regimen as that of patient 1. The esophageal tumor was reduced, and a curative esophagectomy was performed. The patient is now alive without recurrence 38 months after chemotherapy. In conclusion, preoperative chemotherapy using a combination of 5-FU and CDDP may thus be an effective treatment for patients with advanced BSC of the esophagus. PMID- 12768372 TI - Long-term survival following treatment with antineoplastons for colon cancer with unresectable multiple liver metastases: report of a case. AB - We report a case of survival for nearly 8 years after treatment of unresectable multiple liver metastases from colon cancer, using microwave ablation and the nontoxic antitumor agent, the antineoplastons. A 72-year-old man diagnosed with adenocarcinoma of the ascending colon and 14 bilateral liver metastases underwent a right hemicolectomy combined with microwave ablation of six metastatic liver tumors. We also decided to give antineoplastons to inhibit metastatic tumor growth and recurrence. Antineoplaston A10 was given intravenously, followed by oral antineoplaston AS2-1. Computed tomography scans done 1 and 4 years after the initial diagnosis showed recurrent tumors in S(4) and S(7), respectively. The patient underwent a second and a third microwave ablation of the recurrent tumors, and has survived for nearly 8 years without suffering any serious adverse effects. He is currently free from cancer. This case report demonstrates the potential effectiveness of the nontoxic antitumor agent, the antineoplastons, for controlling liver metastases from colon cancer. PMID- 12768373 TI - Rectal malignant melanoma diagnosed by N-isopropyl- p-123I-iodoamphetamine single photon emission computed tomography and 5-S-cysteinyl dopa: report of a case. AB - Malignant melanoma in the anorectal region is a rare disease associated with a very poor prognosis. Taking a biopsy of malignant melanoma is generally contraindicated because of the high risk of inducing metastasis. Although clinical examination and imaging findings are important for the preoperative diagnosis, conventional imaging techniques sometimes fail to provide information from which an accurate diagnosis can be made. We recently treated an 84-year-old woman with rectal malignant melanoma, in whom magnetic resonance imaging showed atypical findings. On the other hand, N-isopropyl- p-(123)I-iodoamphetamine single photon emission computed tomography and 5- S-cysteinyl dopa in blood serum, as a tumor marker of malignant melanoma, proved very effective for establishing the preoperative diagnosis. Despite radical abdominoperineal resection, the patient died of multiple liver and lung metastases about 5 months after surgery. PMID- 12768374 TI - Hemoperitoneum from a spontaneous rupture of a giant hemangioma of the liver: report of a case. AB - Hemangioma is the most common benign tumor of the liver and it is often asymptomatic. Spontaneous or traumatic rupture, intratumoral bleeding, consumption coagulopathy, and rapid growth are mandatory surgical indications. We report a case of giant hemangioma of hepatic segments II and III, which presented as hemoperitoneum, and were treated successfully with preoperative transcatheter arterial embolization (TAE) and hepatic bisegmentectomy. A PubMed Medline search has identified up to now 32 cases of spontaneous rupture of hepatic hemangioma in adults (age >14 years) without a history of trauma, including the present case. Twenty-seven out of these were reviewed. Sixteen (84.2%) of 19 tumors of known size were giant hemangiomas (mean diameter 14.8 cm; range 6-25). Twenty-two (95.7%) patients underwent surgery. Thirteen patients (59.1%) had a resection, 5 (22.8%) were sutured, and 4 (18.1%) underwent tamponade. Three (23%) out of the 13 resected patients died. Four patients (30.8%) underwent TAE prior to elective hepatic resection without any operative mortality. Among the 5 sutured patients, 2 (40%) died as well as 3 (75%) out of 4 patients who underwent tamponade. The mortality rate of all surgery patients was 36.4% (8/22). PMID- 12768375 TI - Primary rhabdoid tumor of the gallbladder: report of a case. AB - A 46-year-old man was operated on for cholelithiasis with chronic cholecystitis. On gross inspection of the resected gallbladder, a slight thickening in the body wall, in an area measuring about 1 x 0.5 cm, was noted. On light microscopic and immunohistochemical examinations, the lesion was diagnosed to be a rhabdoid tumor. After a thorough review of the literature we failed to find any reference to such a lesion in the gallbladder. This is the first known case report of a rhabdoid tumor of the gallbladder. PMID- 12768376 TI - Signet-ring cell carcinoma of the ampulla of Vater: report of a case. AB - An 83-year-old man was admitted to our hospital complaining of general fatigue, fever, and obstructive jaundice. Percutaneous transhepatic bile duct drainage was performed. Gastroduodenal fiberscopy revealed carcinoma of the ampulla of Vater, and early gastric cancer was suspected. A pancreatoduodenectomy with lymph node dissection was performed. Although a biopsy specimen from the gastric lesion was suspected to be well-differentiated adenocarcinoma, no cancerous lesion was found in a specimen resected from the stomach. The histopathologic findings of the ampullary lesion were compatible with a diagnosis of signet-ring cell carcinoma. This is a rare lesion, and a review of the literature revealed only three previous similar cases. PMID- 12768377 TI - Clear cell endocrine tumor of the pancreas which is not associated with von Hippel-Lindau disease: report of a case. AB - We describe the case of a pancreatic endocrine tumor with marked clear cell changes. The patient, a 56-year-old woman, presented with appetite loss and nausea but no evidence of either von Hippel-Lindau disease, or a family history of the disease. A radiological examination revealed a hypervascular tumor measuring 5.2 cm in diameter located in the pancreatic head. The patient underwent a pylorus-preserving pancreatoduodenectomy. The cut surface of the tumor was yellowish and well demarcated. Histologically, the tumor was composed of cuboidal cells with clear cytoplasm, thus forming trabecular and insular nests. Immunohistochemically, the tumor cells were strongly positive for chromogranin. An electron microscopic examination revealed the tumor cells to contain neuroendocrine secretory granules, abundant lipid droplets, and some crystalloid bodies in the cytoplasm. We conclusively diagnosed it to be a pancreatic clear cell endocrine tumor. Such tumors not associated with von Hippel Lindau disease are extremely rare. PMID- 12768378 TI - Spontaneous hematoma of the lateral abdominal wall caused by a rupture of a deep circumflex iliac artery: report of two cases. AB - Expanding hematoma of the abdominal wall is a rare example of acute abdominal disease. We report two cases of lateral abdominal wall hematoma caused by the rupture of a deep circumflex iliac artery, which is a rare cause of an abdominal wall hematoma. Both patients experienced severe abdominal pain after sneezing or coughing. In both cases, computed tomography (CT) findings suggested that active bleeding was continuing. Emergent angiography was therefore performed, and the hematoma was embolized using Spongel or Microcoils. Ultrasound examinations were repeatedly used to monitor the size of the hematoma. The size of the hematoma and patient's pain gradually decreased after embolization. Ultrasound and CT examinations provided useful information for the differential diagnosis of this disease. We conclude that emergent angiography should be performed to control bleeding and avoid any unnecessary surgical procedures in patients with hematoma of the abdominal wall. PMID- 12768379 TI - Malignant hemangiopericytoma in the pelvic cavity successfully treated by combined-modality therapy: report of a case. AB - A 55-year-old Japanese woman underwent extirpation of a malignant hemangiopericytoma in the pelvic cavity, followed by postoperative irradiation. An abdominal computed tomography scan 3 years later revealed a local recurrent tumor, 12 cm in diameter, in the pelvic cavity, for which transarterial embolization was done, followed by excision of the tumor employing Hartmann's procedure. Although an unresectable part of the recurrent tumor remained, postoperative irradiation reduced its size remarkably. The patient is still alive 7 years 2 months after her first operation, but with more recurrent tumors in the abdominal wall and around the bilateral iliac arteries. Because hemangiopericytoma often recurs or metastasizes after a prolonged disease-free interval, close long-term follow-up is necessary after the operation. Combined modality therapy against the recurrent or unresectable disease may result in a good prognosis. PMID- 12768380 TI - Anterior lumbar interbody fusion: does stable anterior fixation matter? AB - The purpose of this study was to compare the outcome of anterior lumbar interbody fusion without instrumentation (uninstrumented ALIF) against that with stable anterior cage fixation using Hartshill horseshoe instrumentation (ALIF-HH) for similar severity of disc disease. Between April 1994 and June 1998 the senior author N.R.B. performed 29 instrumented ALIF procedures with a Hartshill horseshoe cage (ALIF-HH). Between 1990 and 1998, the other senior author (J.M.H.), together with another senior consultant orthopaedic surgeon, performed 27 noninstrumented ALIF procedures using corticocancellous iliac crest autograft. All the patients in both groups had single-level fusion. An independent assessor (S.M.) performed the entire review. The mean follow-up was 4.7 years (2.3-7.9 years) in the uninstrumented ALIF group and 3.0 years (2.1-4.4 years) in the ALIF HH group. There was subsidence of graft in four patients in the uninstrumented ALIF group. It is reasonable to assume that there was no pseudarthrosis in the ALIF-HH group. This difference was statistically significant (two-sided P-value =0.0425). On subjective score assessment, there was a satisfactory outcome (score0.05). On classification by the Oswestry Index into four categories, we found no difference in outcome between the two groups: 83.3% ( n=20) had a satisfactory outcome (defined as Excellent or Better) with ALIF and 77.8% ( n=21) had a satisfactory outcome with ALIF-HH using the Oswestry Disability Index for post-operative assessment ( P>0.05). The results of this study indicate that the Hartshill horseshoe cage does improve the fusion rate, but does not affect clinical outcome. PMID- 12768381 TI - Spinal stenosis surgery in Sweden 1987-1999. AB - Despite being recognised for many years as a clinical diagnosis, no exact definition of spinal stenosis has yet been agreed, leading to difficulties in interpreting and comparing studies of the incidence, prevalence and treatment. This study presents the first analysis of national data to be reported. It is a retrospective population-based national register study, aimed at analyzing surgical interventions in patients with lumbar spinal stenosis, patient characteristics, subsequent development, and case fatality rate, based on Swedish national data for 1987-1999. Complete follow-up data were obtained of incidence and type of spinal stenosis surgery, rate of multiple operations, mortality, underlying causes of death, length of hospital stay, and case fatality rate by linkage of the National Inpatient Register and Swedish Death Register. The study cohort consisted of 10,494 patients. Laminectomy was performed in 89%, and additional fusion in 11%. The mean annual rate of operations was 9.7 per 100,000 inhabitants, the annual number of operations performed increased from 4.7 to 13.2 per 100,000 inhabitants per year. The case fatality rate within 30 days after surgery was 3.5 per 1000 operations. Cardiovascular disease was the most common cause of death (46%). Relative risk of dying within 30 days of admission was doubled in men, and for fusion surgery, and increased four fold in patients older than 80 years. The relative risk of dying decreased during the study period. The results show that spinal stenosis surgery in Sweden has increased, and is associated with a low risk. Within an ageing group of patients, mortality has declined. PMID- 12768382 TI - Diversity of arbuscular mycorrhizal fungi colonising roots of the grass species Agrostis capillaris and Lolium perenne in a field experiment. AB - Analysis of arbuscular mycorrhizal (AM) fungal diversity through morphological characters of spores and intraradicular hyphae has suggested previously that preferential associations occur between plants and AM fungi. A field experiment was established to investigate whether AM fungal diversity is affected by different host plants in upland grasslands. Indigenous vegetation from plots in an unimproved pasture was replaced with monocultures of either Agrostis capillaris or Lolium perenne. Modification of the diversity of AM fungi in these plots was evaluated by analysis of partial sequences in the large subunit (LSU) ribosomal RNA (rDNA) genes. General primers for AM fungi were designed for the PCR amplification of partial sequences using DNA extracted from root tissues of A. capillaris and L. perenne. PCR products were used to construct LSU rDNA libraries. Sequencing of randomly selected clones indicated that plant roots were colonised by AM fungi belonging to the genera Glomus, Acaulospora and Scutellospora. There was a difference in the diversity of AM fungi colonising roots of A. capillaris and L. perenne that was confirmed by PCR using primers specific for each sequence group. These molecular data suggest the existence of a selection pressure of plants on AM fungal communities. PMID- 12768383 TI - Pathophysiological significance of neuronal nitric oxide synthase in the gastrointestinal tract. AB - It has been demonstrated that nitric oxide (NO) is a major inhibitory nonadrenergic, noncholinergic (NANC) neurotransmitter in the gastrointestinal (GI) tract. NO released in response to nerve stimulation of the myenteric plexus causes relaxation of the smooth muscle. NO is synthesized by the activation of neuronal NO synthase (nNOS) in the myenteric plexus. Released NO plays an important physiological role in various parts of the GI tract. NO regulates the muscle tone of the sphincter in the lower esophagus, pylorus, sphincter of Oddi, and anus. NO also regulates the accommodation reflex of the fundus and the peristaltic reflex of the intestine. Previous studies have shown that NOS inhibitors delay gastric emptying and colonic transit. The reduction of nNOS expression, associated with impaired local production of NO, may be responsible for motility disorders in the GI tract. There is accumulated evidence that dysfunction of NO neurons in the myenteric plexus may cause various GI diseases. These reports are reviewed and possible mechanisms of altered nNOS expression are discussed in this article. In particular, impaired nNOS synthesis of the myenteric plexus seems to be an important contributing factor to the pathogenesis of achalasia, diabetic gastroparesis, infantile hypertrophic pyloric stenosis, Hirschsprung's disease, and Chagas' disease. Reduced NO release and/or nNOS expression are suspicious in a subset of patients with functional dyspepsia. Although the etiology of intestinal pseudo-obstruction remains unknown, it is conceivable that extrinsic denervation may upregulate nNOS expression, resulting in enhanced muscular relaxation and disturbed peristalsis. An animal model of colitis showed impaired nNOS expression in the colonic myenteric plexus. Antecedent infection may be associated with the impaired NO pathways observed in functional dyspepsia, colitis, and Chagas' disease. PMID- 12768384 TI - Expression of Tie-2 and angiopoietin-1 and -2 in early phase of ulcer healing. AB - BACKGROUND: Angiogenesis is an important process in tissue development and wound healing. The Tie-2 receptor tyrosine kinases and ligands, angiopoietin (Ang)-1 and -2 have been postulated to play key roles in vascular development. The purpose of this study was to evaluate the expression of Tie-2 and Ang-1 and -2 in an acetic acid-induced gastric ulcer healing process in rats. METHODS: Gastric specimens were obtained at 0 (control), 1, 3, 5, 7, and 14 days after ulcer induction for reverse transcription polymerase chain reaction (RT-PCR), Western blot analysis, and immunohistochemical analysis. RESULTS: Expression of Tie-2 and Ang-1 and -2 mRNAs was detected in normal gastric tissue and ulcerative tissues by RT-PCR. Western blot analysis revealed that Tie-2 expression reached a maximum on the third to fifth days. Expression of Ang-1 and -2 peaked on the first day. Ang-1 expression gradually became weaker in 2 weeks, whereas Ang-2 expression returned to normal in a few days. Immunohistochemically, Tie-2 was expressed constitutively in the endothelial cells of pre-existing vessels of the gastric wall, and Tie-2 expression was increased in the new capillaries of the ulcer base. CONCLUSIONS: These findings suggest that Tie-2 and Ang-1 and -2 play an important role in angiogenesis in the early phase of ulcer healing. PMID- 12768385 TI - Changing pattern of antimicrobial resistance of Helicobacter pylori in Korean patients with peptic ulcer diseases. AB - BACKGROUND: Antibiotic resistance of Helicobacter pylori is problematic because it reduces the efficacy of eradication therapy. It has been suggested that the incidence of resistance is rising. In Korea, information on the antimicrobial resistance of H. pylori is rare. The aim of this study was to assess the prevalence of H. pylori antibiotic resistance at a single center in Korea, and the changes in its antimicrobial resistance, and to detect the mutation foci of clarithromycin-resistant strains. METHODS: H. pylori isolates obtained from 224 patients with peptic ulcer disease in Korea between June 1996 and March 2000 were tested for antimicrobial resistance. The minimum inhibitory concentration (MIC) for metronidazole and clarithromycin was determined by the broth microdilution method. Isolates were considered resistant when the MIC was more than 8 microg/ml for metronidazole and more than 1 microg/ml for clarithromycin. To detect H. pylori 23S rRNA mutations, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was performed. Sequencing was performed on the two strands of the nonrestricted amplicons. RESULTS: Overall, resistance to metronidazole and clarithromycin was detected in 41.9% and 5.4% of patients, respectively. There was no significant difference in metronidazole and clarithromycin resistance according to age group and sex. Six strains were resistant to both metronidazole and clarithromycin. Six of nine clarithromycin resistant isolates possessed the A2144G mutation in the gene encoding 23S rRNA. Sequencing of the three non-restricted clarithromycin-resistant strains revealed a T-to-C mutation at position 2182. CONCLUSIONS: In Korea, there was no significant increase in the prevalence of metronidazole resistance, but clarithromycin-resistant H. pylori strains had increased relatively over the 5 year period. There was an increasing tendency for the emergence of strains with dual resistance to metronidazole and clarithromycin. Many of the clarithromycin resistant strains possessed the A2144G mutation. PMID- 12768386 TI - Reorganized small intestine from fetal mouse as an in vitro wound healing model. AB - BACKGROUND: We developed a method for reorganizing the mouse small intestine. In the present study, we investigated whether the reorganized small intestine was morphologically and histochemically differentiated. We also evaluated the reorganized small intestine as an in vitro wound healing model. METHODS: Fetal mouse small intestines were dispersed into single cells, which were then cultured to a high density. Newly formed small intestine-like organs on a membrane filter were observed by light and electron microscopy. Alkaline phosphatase (ALPase) activity of the epithelium was analyzed. To evaluate the reorganized small intestine as an in vitro wound healing model, a scalpel was used to cut the reorganized intestine on a membrane, and the healing process was morphologically and immunohistochemically examined. RESULTS: After 6 days in culture, the surface was almost completely coveed with epithelial cells, and villus-like structures were observed. These epithelial cells formed microvilli, and in parallel with this development, ALPase activity of the microvilli increased (from day 4). Twenty-four hours after the cutting, the wound surface was almost completely covered with undifferentiated epithelial cells. The number of acetylated low density lipoprotein labeled with 1,1,dioctadecyll,3,3,3,3, tetramethyl indocarbocyanine perchlorate (DiI-Ac-LDL)-positive macrophages increased after cutting. Platelet-derived growth factor (PDGF)-, basic fibroblast growth factor (bFGF)-, matrix metalloproteinase-1 (MMP-1)-positive cells were detected by immunohistochemical staining. CONCLUSIONS: The reorganized small intestine had a morphologically and histochemically differentiated organoid structure, and was useful as an in vitro model for investigating the process of wound healing. PMID- 12768387 TI - A novel diamino-pyridine derivative (IS-741) attenuates rat ileitis induced by trinitrobenzene sulfonic acid. AB - BACKGROUND: The etiology and pathogenesis of inflammatory bowel disease remain unknown. However, neutrophil infiltration into the inflammatory lesion is an important process in inflammatory bowel disease. In this study, we used rat trinitrobenzene sulfonic acid (TNBS) ileitis as a Crohn's disease model, and investigated the effects of oral IS-741 (which inhibits the expression of Mac-1, a cell adhesion molecule) on leukocyte-endothelial interactions. METHODS: Rat ileitis was induced by the intraluminal injection of a TNBS solution (160 mg/kg in 50% ethanol) at a site 10 cm proximal to the ileocecal valve. The rats then received oral IS-741 (50 mg/kg) or saline for 7 days. On the day 8 after the initial administration of IS-741 or saline, we determined the visible damage score, and assessed myeloperoxidase (MPO) activity. Concentrations of cytokines in the ileum, such as interleukin-8 (IL-8) and tumor necrosis factor-alpha (TNF alpha) were assayed by enzyme-linked immunosorbent assay (ELISA). We also investigated the infiltration of polymorphonuclear cells and Mac-1 positive cells by histological examinations. RESULTS: The administration of IS-741 resulted in a significant reduction of the visible damage score, myeloperoxidase (MPO) activity, and mucosal IL-8 levels in the ileum as compared with the saline administration. IS-741 also dramatically reduced the infiltration of polymorphonuclear cells and Mac-1 positive cells into the inflamed lesions. CONCLUSIONS: These results indicate that the oral administration of IS-741 inhibits neutrophil infiltration into inflamed lesions, and is effective for attenuating rat TNBS ileitis. This new anti-inflammatory agent may be beneficial for the treatment of inflammatory bowel disease. PMID- 12768388 TI - Relaxation effects of adrenomedullin in carbachol-precontracted rabbit internal anal sphincter. AB - BACKGROUND: The internal anal sphincter is a specialized continuation of the circular smooth muscle layer of the rectum. It has been found that internal anal sphincter tonus is significantly increased in the majority of patients with anal fissure. Adrenomedullin has a relaxant effect on smooth muscle in different kinds of tissue. In this in vitro study, we aimed to investigate whether adrenomedullin had a relaxant effect on isolated rabbit internal anal sphincter, and the possible involvement of nitric oxide, prostaglandins, and K(+) channels. METHODS: Internal anal sphincter smooth muscle strips obtained from New Zealand rabbits were studied in vitro. The effects of adrenomedullin were investigated in isolated strips of rabbit internal anal sphincter smooth muscle precontracted with carbachol alone, and in the presence of indomethacin (a cyclooxygenase inhibitor), N(omega)-nitro- l-arginine methyl ester ( l-NAME; a nitric oxide synthetase inhibitor), and K(+) channel blockers. RESULTS: Adrenomedullin caused relaxation of the isolated precontracted rabbit internal anal sphincter strips in a concentration-dependent manner. The response of the internal anal sphincter was not affected by l-NAME, indomethacin, and the K(+) channel blockers. CONCLUSIONS: This relaxant effect of adrenomedullin might lead to novel clinical applications of adrenomedullin for anorectal disorders with increased internal anal sphincter tonus, such as anal fissure. PMID- 12768389 TI - Acute exacerbation during interferon alfa treatment of chronic hepatitis B: frequency and relation to serum beta-2 microglobulin levels. AB - BACKGROUND: We aimed to determine the frequency of alanine aminotransferase (ALT) elevation during interferon-alpha treatment, the so-called "flare", its relation to serum beta-2 microglobulin levels, and its impact on the outcome of treatment in chronic hepatitis B. METHODS: The files of 53 treatment-naive patients with chronic hepatitis B (17 hepatitis B e antigen (HBeAg) +ve, 36 HBeAg -ve) who had been treated with 10 MU interferon-alpha 2b three times per week for 24 weeks were reviewed. We analyzed the fluctuations in serum ALT, beta(2)-microglobulin, and HBV-DNA levels before, during, and after flare. RESULTS: We detected flare in 4/17 (24%) of the HBeAg +ve and 7/34 (21%) of the HBeAg -ve patients. ALT level peaked between weeks 2 and 16 (mean, week 8). After flare, HBV-DNA disappeared in 5/7 (71%) HBeAg -ve vs 3/4 (75%) HBeAg +ve patients (all seroconverted to anti HBe). The overall sustained response rate was 41%: 55% in the patients with flare, and 38% in those without ( P > 0.05). Basal serum beta(2)-microglobulin levels were significantly higher in responders vs nonresponders (2.19 +/- 0.32 vs 1.78 +/- 0.34 mg/l, mean +/- SD; P < 0.005). In addition, during treatment, serum beta(2)-microglobulin levels increased significantly only in responders, and the degree of increase was significantly higher in responders with flare vs responders without flare (3 +/- 0.33 vs 2.34 +/- 0.35 mg/l; P < 0.001). CONCLUSIONS: This study, with a limited sample size, showed that, in chronic hepatitis B, there is a trend for a higher response in patients with exacerbation of hepatitis B with interferon-alpha treatment. However, the difference does not reach statistical significance to be of predictive value. On the other hand, serum beta(2)-microglobulin levels before and during treatment may be useful in predicting the outcome. PMID- 12768390 TI - Serum leptin level can be a negative marker of hepatocyte damage in nonalcoholic fatty liver. AB - BACKGROUND: The aim of this study was to determine whether leptin and insulin resistance (IR) showed differences between steatotic patients with and without elevated serum transaminases. METHODS: The study included 32 patients with fatty liver and high serum transaminase level (group I), 31 patients with fatty liver and normal serum transaminase level (group II), and 8 nonobese and nonsteatotic controls. The presence of steatosis was demonstrated by ultrasonography. Due to the effect of body mass index (BMI) on leptin levels, groups I and II were divided to form four subgroups for analysis (group IA, BMI 30; group IIA, BMI 30. RESULTS: The serum leptin levels in group IIB were significantly higher than the levels in group IB ( P = 0.017). Serum leptin was also higher in group IIA than in group IA, but this difference was not statistically significant ( P = 0.097). Logistic regression analysis revealed a significant negative correlation between serum leptin level and the presence of a high transaminase level (odds ratio, 0.97; 95% confidence interval, 0.95-0.99). The levels of IR in the four patient groups were comparable, but the controls had significantly lower IR levels than group IIA. CONCLUSIONS: Elevated serum leptin seems to be a feature of steatotic patients with normal transaminase levels, and the level of serum leptin seems to decrease as the hepatocyte injury develops. IR is a common feature of fatty liver disease, irrespective of the presence of hepatocellular necrosis. PMID- 12768391 TI - Esophageal tuberculosis presenting with an appearance similar to that of carcinoma of the esophagus. AB - A case of esophageal tuberculosis presenting with an appearance similar to that of esophageal cancer is reported. The patient was an 82-year-old man with progressive dysphagia. Barium swallow and esophagoscopy revealed an elevated lesion with deep ulceration in the middle thoracic esophagus. Esophageal carcinoma, in particular, an undermining type of undifferentiated carcinoma, was suspected fluoroscopically and endoscopically. Histological examination of biopsy specimens revealed no malignancy, but there were epithelioid granulomas and a few Langhans' type multinucleated giant cells. Endoscopic ultrasonography clearly demonstrated an extramural lesion with calcification and direct infiltration of enlarged subcarinal lymph nodes into the esophageal wall. Ultrasonographic and histological findings indicated the possibility of esophageal tuberculosis. Although no bacteriological evidence was obtained, a therapeutic trial for tuberculosis, using antituberculous drugs, was started. After 2 weeks, the enlarged subcarinal lymph nodes were markedly reduced in size. The patient's symptoms improved gradually and had disappeared 8 weeks after he started treatment, when tubercle bacilli were isolated from sputum. A connection between the esophageal wall and its adjacent structures was clearly demonstrated by endoscopic ultrasonography. For patients with findings indicative of esophageal tuberculosis on endoscopic ultrasonography, a therapeutic trial for tuberculosis should be considered, even if polymerase chain reaction assay or culture is negative. PMID- 12768392 TI - Concomitant colitis associated with primary sclerosing cholangitis. AB - Between 1985 and 2001, seven Japanese patients (four males and three females) were diagnosed as having primary sclerosing cholangitis (PSC) in our hospital. All seven patients received total colonoscopy with distal ileoscopy. All four male patients were diagnosed as having colitis by colonoscopy, while none of the three female patients had colitis. The four patients with colitis did not have any symptoms attributable to colitis, such as diarrhea or hematochezia. In three of the four patients, unclassified colitis was the most suitable diagnosis, because there were no typical findings of ulcerative colitis or Crohn's disease. The remaining patient was diagnosed as having eosinophilic colitis. By colonoscopic visualization, the right-sided colon, including the terminal ileum, was mainly involved, but the lesions were not severe. The main findings were redness, erosion, stenosis, and insufficiency of haustral formation. Histologically, these lesions were nonspecific inflammatory changes in the three patients with unclassified colitis. In the patient with eosinophilic colitis, remarkable infiltration of eosinophils was observed. Thus, unclassified colitis appeared to be the main complication in these patients with PSC. Males predominated in regard to concomitant colitis, and they had no symptoms of the colitis. Colonoscopic examination revealed that the lesions were not severe. The main lesions were found in the right-sided colon, with nonspecific inflammatory changes. These results suggest that colonoscopic surveillance of patients with PSC should be performed even if they do not have any colitis symptoms. PMID- 12768393 TI - A 25-year clinical history of portopulmonary hypertension associated with latent myeloproliferative disorder. AB - Pulmonary hypertension associated with increased pulmonary vascular resistance occurring in the setting of portal hypertension, referred to as "portopulmonary hypertension", is a complication of chronic liver disease, and occurs in 2% to 3% of patients with portal hypertension. Portal hypertension is a relatively common finding in patients with chronic myeloproliferative disorder (CMPD). Pulmonary hypertension is also an occasional finding in CMPD patients. Latent myeloproliferative disorder, on the other hand does not fulfill the diagnostic criteria of classical CMPD and is characterized by younger age of onset, slow disease progression, a high risk of thrombosis, platelet dysfunction, and normal or increased platelet count in spite of the presence of splenomegaly. We report findings in a 50-year-old woman with portal hypertension for which there were three major etiological findings-increased splenic blood flow, infiltration of hematopoietic cells in the liver, and thrombosis in the portal or hepatic vein over a 25-year clinical course, during which there was also reversible stenosis of the portal vein. Twenty-three years after her first admission, her condition was diagnosed as latent myeloproliferative disorder, and she developed pulmonary hypertension. Her clinical history and data indicated that the portopulmonary hypertension was due to the latent myeloproliferative disorder. PMID- 12768394 TI - Type C-chronic hepatitis patients who had autoimmune phenomenon and developed jaundice during interferon therapy. AB - Of a total of 2342 patients with type-C chronic hepatitis treated with interferon (IFN) at this hospital, 3 patients developed jaundice during the course of IFN therapy, but all 3 of them exhibited negative conversion of hepatitis C virus (HCV)-RNA following readministration of IFN. All 3 patients were assessed as having "probable autoimmune hepatitis (AIH)" in accordance with the AIH scoring system, indicating association with an "autoimmune phenomenon". Readministration of IFN at a low-dose induced activation of the autoimmune phenomenon, leading to fulminant hepatocellular impairment. As a result, HCV-RNA content dropped dramatically, possibly contributing to the negative conversion of HCV-RNA noted following the readministration of IFN. At present, no adequate therapy has been established for type-C chronic hepatitis with autoimmune manifestations. However, in conclusion, our findings suggested that: IFN should be a frontline regiment: (1). if autoantibody titers are low or patients are rated as having "probable AIH" or lower in accordance with the AIH scoring system, indicating a strong link to chronic hepatitis or (2). if IFN is expected to be efficacious on the basis of genotype of HCV-RNA level; even if there is acute exacerbation of type-C chronic hepatitis, IFN should be re-administered in the HCV-RNA level has dropped subsequently. PMID- 12768395 TI - Serous cystadenoma of the pancreas associated with obstructive jaundice. AB - We herein report a case of pancreatic serous cystadenoma in a patient who presented with jaundice, and we provide a review of the literature. A 53-year-old man was admitted with complaints of jaundice and weight loss. With a preoperative diagnosis of pancreatic serous cystadenoma with obstructive jaundice, he underwent pylorus-preserving pancreatoduodenectomy. A cystic tumor partially protruding into the bile duct was observed in the pancreatic head. Histology verified serous cystadenoma of the pancreas. Histologically, no atypia was proven in the epithelium. There have been only eight case reports dealing with serous cystadenoma of the pancreas with obstructive jaundice. Although serous cystadenoma of the pancreas has essentially a benign nature, pylorus-preserving pancreatoduodenectomy is the treatment of choice when available to avoid the recurrence of obstructive jaundice. PMID- 12768396 TI - Hemobilia caused by liver abscess due to intrahepatic duct stones. AB - Hemobilia occurs when injury or disease causes communication between intrahepatic blood vessels and the biliary tract. Causes of hemobilia include trauma; gallstones; inflammatory diseases; and vascular disorders such as aneurysm, tumor, and coagulopathy. Recently, with the increasing use of invasive diagnostic and therapeutic procedures involving the hepatobiliary tract, an increasing proportion of the causes of hemobilia have been of iatrogenic origin. Hemobilia may also be associated with liver abscess, but this condition is very rare. Our review of the English-language literature disclosed few cases of liver abscess associated with hemobilia. Here, we present a case of hemobilia caused by liver abscess due to intrahepatic duct stones. Liver abscess should be considered in the causes of hemobilia, especially in areas where hepatobiliary parasitic infection is endemic. PMID- 12768397 TI - A case of primary follicular lymphoma of the duodenum with BCL-2 gene rearrangement. PMID- 12768398 TI - Prevalence of reflux esophagitis in patients with duodenal ulcer and gastric ulcer. PMID- 12768399 TI - Hepatocellular adenoma presenting as a giant multicystic tumor of the liver. PMID- 12768400 TI - Reality and false premises of antimicrobial susceptibility testing for Helicobacter pylori. PMID- 12768401 TI - "Insufficient" leptin production for the fat mass: a risk factor for nonalcoholic steatohepatitis in obese patients? PMID- 12768402 TI - Pseudomembranous and neutropenic enterocolitis in pediatric oncology patients. AB - Neutropenic enterocolitis in oncological patients represents a wide spectrum of clinicopathological pictures each with its own entity. Early diagnosis of enterocolitis can lead to improved supportive care and therefore better outcome. We present two cases--patient A, a child with pseudomembranous colitis caused by Clostridium difficile, and patient B, a child with neutropenic enterocolitis, where no organism was found. By allowing an insight into the pathology, immunology and culture results, we demonstrated that early diagnosis leads to improved management and therefore improved outcome. PMID- 12768403 TI - Scalp hypothermia to prevent chemotherapy-induced alopecia is effective and safe: a pilot study of a new digitized scalp-cooling system used in 74 patients. AB - GOALS: The aim of this study was to examine the efficacy and safety of a new digitized, controlled, scalp-cooling system to prevent chemotherapy-induced alopecia. METHOD: Seventy-four female cancer patients who received 13 varying chemotherapy regimens were included in a nonrandomized pilot study. The Digni 2-3 with Dignicap system consists of a refrigerator unit and a control unit integrated into a mobile cabinet and connected to a tight-fitting cooling cap. This system maintains a constant scalp temperature of +5 degrees C for many hours. In this study, 60 patients were treated for ovarian cancer with either taxane or epirubicin combination chemotherapy. Eight patients with Hodgkin's lymphoma, three with breast cancer, two with endometrial cancer, and one with sarcoma were also included. Photo documentation and patient assessment of hair loss and discomfort were performed. RESULTS: In anthracycline-treated patients, total prevention of hair loss was observed, whereas hair loss in paclitaxel/docetaxel-treated patients was minimal to none. The combination of anthracycline and taxane resulted in more hair loss, but only three of six patients used a wig. Scalp cooling was generally very well tolerated; only two of 74 patients discontinued use of the cold cap due to discomfort. No scalp metastases occurred over a median follow-up period of 15 months. CONCLUSIONS: The digitized, controlled, scalp-cooling system represents an effective and safe device that should be clinically evaluated in a randomized trial and in studies using other chemotherapy regimens to determine optimal temperatures and durations of cooling for maximal efficacy. PMID- 12768404 TI - Uncoupling of omnivore-mediated positive and negative effects on periphyton mats. AB - The riverine grass shrimp (Palaemonetes paludosus) and eastern mosquitofish (Gambusia holbrooki) consume periphyton and small invertebrates, potentially affecting periphyton through negative effects (i.e., consumption) and/or positive effects such as nutrient regeneration, physical stimulation, and trophic cascades. We performed field experiments in the Everglades in which omnivores and periphyton were maintained in cages, with a fraction of the periphyton held in omnivore-exclusion bags that allowed passage of nutrients but prevented its consumption or physical disturbance. In some instances, periphyton growth rate increased with increasing omnivore biomass. Omnivores probably stimulated periphyton growth through nutrient regeneration, possibly subsidizing periphyton with nutrients derived from ingested animal prey. The net balance of omnivore mediated negative and positive effects varied among experiments because of seasonal and spatial differences in periphyton characteristics. Consumption of periphyton mats might have been reduced by the arrangement of palatable algae (green algae and diatoms) within a matrix of unpalatable ones (CaCO(3)-encrusting filamentous cyanobacteria). In a laboratory feeding experiment, mosquitofish consumed more green algae and diatoms in treatments with disrupted mat structure than in those with intact mats. No difference in diet was observed for shrimp. Our study underscores the complexity of consumer-periphyton interactions in which periphyton edibility affects herbivory and consumers influence periphyton through multiple routes that cannot be fully appreciated in experiments that only investigate net effects. PMID- 12768405 TI - Positive and negative effects of leaf shelters on herbivorous insects: linking multiple herbivore species on a willow. AB - We experimentally examined the effects on other herbivorous insects of leaf shelters constructed by lepidopteran larvae on a willow, Salix miyabeana. Several insect species occupied the vacant leaf shelters. Our experiment using artificial leaf shelters showed that the number of aphids increased with the number of artificial leaf shelters on a shoot, as did the numbers of three ant species ( Camponotus japonicus, Lasius hayashi, and Myrmica jessensis) that entered leaf shelters to collect aphid honeydew. To determine the ant-mediated effect of leaf shelters on herbivorous insects that do not use leaf shelters, we transferred newly hatched larvae of a common leaf beetle, Plagiodera versicolora, to the leaves of shoots with and without artificial leaf shelters. One day after the transfer, larval survival rate was significantly lower on shoots with shelters than on those without shelters, and shoots with shelters had significantly more ants than did shoots without shelters. Our field experiments demonstrated clearly that shelter-making lepidopteran larvae increased the abundance of both aphids and ants and decreased the survival rate of leaf beetle larvae, probably because the larvae were removed by ants that were attracted to the leaf shelters by the aphid colonies. PMID- 12768406 TI - Sociality and individual fitness in yellow-bellied marmots: insights from a long term study (1962-2001). AB - Theoretical and empirical studies suggest that the age of first reproduction (the age at which reproduction begins) can have a substantial influence on population dynamics and individual fitness. Using complete survival and reproductive histories of 428 female yellow-bellied marmots (Marmota flaviventris) from a 40 year study (1962-2001), we investigated causes and fitness consequences of delayed maturity. Most females (86%) died without reproducing. The age of first reproduction of females that survived to reproduce at least once (n=60) ranged from 2 to 6 years. Females maturing later did not have a larger lifetime number of successful reproductive events or offspring production, nor did they experience improved survival. Females reproducing earlier had a higher fitness than those that delayed maturity. These results suggest that the net cost of early maturity was less than fitness benefits associated with early onset of reproduction, and that age of first reproduction in our study population is under substantial directional selection favoring early maturity. We conclude that female yellow-bellied marmots delay onset of reproduction not because of fitness benefits of foregoing reproduction at an earlier age, but due to the social suppression of reproduction by older, reproductive females, which enhances their own fitness to the detriment of the fitness of young females. Our results indicate that female yellow-bellied marmots that survive to reproduce may act to increase their own direct fitness, and that social suppression of reproduction of young females is a part of that strategy. PMID- 12768408 TI - Distribution and chemical coding of cocaine- and amphetamine-regulated transcript peptide (CART)-immunoreactive neurons in the guinea pig bowel. AB - Cocaine- and amphetamine-regulated transcript-immunoreactive (CART-IR) neurons and nerve fibers were abundant in the submucosal and myenteric plexuses of the guinea pig duodenum, ileum, cecum, proximal and distal colon. CART immunoreactivity was also observed in cell bodies and nerve fibers in the extrahepatic biliary tract. In the myenteric plexus, similar proportions (~20 25%) of neurons were CART-IR in all regions, with the exception of the cecum, where only 13% were CART-IR. In the submucosal plexus, CART-IR was detected in 35 50% of the neurons along the bowel with the exception of the proximal colon (~10%). Multiple label immunohistochemistry in the myenteric plexus of the ileum demonstrated that CART-IR neurons were also immunoreactive for choline acetyltransferase (83%), tachykinins (77%), calbindin (50%), nitric oxide synthase (20%), and/or vasoactive intestinal peptide (23%). In triple label studies, we found that ~8% of the CART-IR neurons were also immunoreactive for both choline acetyltransferase and nitric oxide synthase. CART immunoreactivity was not colocalized with calretinin, somatostatin, or serotonin. These results, combined with previous studies of chemical coding and projection patterns in the guinea pig ileum, indicate that at least four different classes of gut neurons in the myenteric plexus express CART peptide, including excitatory and inhibitory motor neurons projecting to the circular muscle, intrinsic primary afferent neurons, and a subset of descending interneurons. Because all CART-IR neurons in the submucosal plexus were also vasoactive intestinal peptide-IR, they are likely to include secretomotor/vasodilator neurons. PMID- 12768407 TI - The blood-brain barrier of the chick glycogen body (corpus gelatinosum) and its functional implications. AB - Among recent vertebrates only birds possess a glycogen body (corpus gelatinosum), located in the rhomboidal sinus of the lumbosacral region of the spinal cord and separated from the neural tissue proper. Because of the specific topographical situation of this circumventricular organ, the structure of its vascular system is of special interest with respect to the still unsolved functional problems. The existence of a blood-brain barrier is demonstrated by the exclusion of intravascularly injected tracer (horseradish peroxidase), and immunocytochemical demonstration of glucose transporter-1 as a functional marker and of neurothelin, occludin and ZO-1 as structural markers. Alkaline phosphatase and gamma glutamyltransferase activities, two enzyme reactions frequently used for demonstration of an established blood-brain barrier in vitro, were localized histochemically on the plasmalemma of glycogen body cells and were absent from the endothelium. In addition, local enlargements of the intercellular space were observed by transmission and scanning electron microscopy. In accordance with the concept of a third circulation the cerebrospinal fluid may be the vehicle for distributing substances originating in the glycogen body to the CNS, while the vascular endothelium maintains the internal milieu by virtue of its dynamic barrier functions. PMID- 12768409 TI - Impact of public health strategies on the birth prevalence of cystic fibrosis in Brittany, France. AB - Taking into account the situation of Brittany, a region of western France where cystic fibrosis (CF) is common and where a neonatal screening program was set up 14 years ago, the aim of this study was to determine the way in which the birth prevalence of CF has been influenced by the various public health strategies implemented in the region (neonatal screening, prenatal diagnosis, ultrasound examination and family testing). This study used the results of the neonatal screening program, which enabled a precise measure of the prevalence of CF at birth to be obtained. Over the same period, we collected data from prenatal diagnoses carried out in the region, first in families related to a CF child and also those made following the detection of an echogenic bowel upon routine ultrasound examination performed during pregnancy. The prevalence of CF at birth was estimated to be 1/2838 in the region over a 10-year period (1992-2001). By including the 54 CF-affected pregnancies that were terminated during these 10 years, the corrected birth prevalence of CF was 1/1972. Prenatal diagnosis was therefore responsible for a global decrease in CF prevalence at birth of 30.5%. This work constitutes the first study able to provide a precise measure of CF birth prevalence and of its evolution through the combined effects of neonatal screening, prenatal diagnosis, ultrasound examination and family testing. PMID- 12768410 TI - Comparison of the utility of barley retrotransposon families for genetic analysis by molecular marker techniques. AB - The Sequence-Specific Amplification Polymorphism (S-SAP) method, and the related molecular marker techniques IRAP (inter-retrotransposon amplified polymorphism) and REMAP (retrotransposon-microsatellite amplified polymorphism), are based on retrotransposon activity, and are increasingly widely used. However, there have been no systematic analyses of the parameters of these methods or of the utility of different retrotransposon families in producing polymorphic, scorable fingerprints. We have generated S-SAP, IRAP, and REMAP data for three barley (Hordeum vulgare L.) varieties using primers based on sequences from six retrotransposon families (BARE-1, BAGY-1, BAGY-2, Sabrina, Nikita and Sukkula). The effect of the number of selective bases on the S-SAP profiles has been examined and the profiles obtained with eight MseI+3 selective primers compared for all the elements. Polymorphisms detected in the insertion pattern of all the families show that each can be used for S-SAP. The uniqueness of each transposition event and differences in the historic activity of each family suggest that the use of multiple retrotransposon families for genetic analysis will find applications in mapping, fingerprinting, and marker-assisted selection and evolutionary studies, not only in barley and other Hordeum species and related taxa, but also more generally. PMID- 12768411 TI - Differential modulation of Bordetella pertussis virulence genes as evidenced by DNA microarray analysis. AB - The production of most factors involved in Bordetella pertussis virulence is controlled by a two-component regulatory system termed BvgA/S. In the Bvg+ phase virulence-activated genes (vags) are expressed, and virulence-repressed genes (vrgs) are down-regulated. The expression of these genes can also be modulated by MgSO(4) or nicotinic acid. In this study we used microarrays to analyse the influence of BvgA/S or modulation on the expression of nearly 200 selected genes. With the exception of one vrg, all previously known vags and vrgs were correctly assigned as such, and the microarray analyses identified several new vags and vrgs, including genes coding for putative autotransporters, two-component systems, extracellular sigma factors, the adenylate cyclase accessory genes cyaBDE, and two genes coding for components of a type III secretion system. For most of the new vrgs and vags the results of the microarray analyses were confirmed by RT-PCR analysis and/or lacZfusions. The degree of regulation and modulation varied between genes, and showed a continuum from strongly BvgA/S activated genes to strongly BvgA/S-repressed genes. The microarray analyses also led to the identification of a subset of vags and vrgs that are differentially regulated and modulated by MgSO(4) or nicotinic acid, indicating that these genes may be targets for multiple regulatory circuits. For example, the expression of bilA, a gene predicted to encode an intimin-like protein, was found to be activated by BvgA/S and up-modulated by nicotinic acid. Furthermore, surprisingly, in the strain analysed here, which produces only type 2 fimbriae, the fim3 gene was identified as a vrg, while fim2 was confirmed to be a vag. PMID- 12768413 TI - Mutations arise independently of transcription in non-dividing bacteria. AB - It has been proposed that transcription introduces a bias into the random process of mutation. Although this hypothesis is supported by experimental data for mutations arising during active bacterial growth, the role of transcription in mutagenesis in non-dividing bacteria is entirely hypothetical. In the present study, we tested the hypothesis of a possible role of transcription in a non dividing E. coli K12 strain. In this strain (BD010), a mutated trpB allele (trpB9578), placed under stringent transcriptional control, was tested for the appearance of prototrophic revertants on synthetic medium lacking tryptophan. The number of phenotypic revertants which appeared in the absence of trp transcription was compared to that observed when the mutated gene was continuously transcribed. Our results showed that transcription of trpB is not mutagenic under conditions of tryptophan starvation, and that the frequency of TrpB+ reversion is solely a function of the duration of starvation. PMID- 12768412 TI - ABC transporters of the wheat pathogen Mycosphaerella graminicola function as protectants against biotic and xenobiotic toxic compounds. AB - We have studied the role of five ABC transporter genes (MgAtr to MgAtr5) from the wheat pathogen Mycosphaerella graminicola in multidrug resistance (MDR). Complementation of Saccharomyces cerevisiae mutants with the ABC transporter genes from M. graminicola showed that all the genes tested encode proteins that provide protection against chemically unrelated compounds, indicating that their products function as multidrug transporters with distinct but overlapping substrate specificities. Their substrate range in yeast includes fungicides, plant metabolites, antibiotics, and a mycotoxin derived from Fusarium graminearum (diacetoxyscirpenol). Transformants of M. graminicola in which individual ABC transporter genes were deleted or disrupted did not exhibit clear-cut phenotypes, probably due to the functional redundancy of transporters with overlapping substrate specificity. Independently generated MgAtr5 deletion mutants of M. graminicola showed an increase in sensitivity to the putative wheat defence compound resorcinol and to the grape phytoalexin resveratrol, suggesting a role for this transporter in protecting the fungus against plant defence compounds. Bioassays with antagonistic bacteria indicated that MgAtr2 provides protection against metabolites produced by Pseudomonas fluorescens and Burkholderia cepacia. In summary, our results show that ABC transporters from M. graminicola play a role in protection against toxic compounds of natural and artificial origin. PMID- 12768415 TI - Investigations on the biology, epidemiology, pathology and control of Tunga penetrans in Brazil. II. Prevalence, parasite load and topographic distribution of lesions in the population of a traditional fishing village. AB - Tungiasis is hyperendemic in many poor communities in Brazil and is associated with considerable morbidity. In order to understand the factors that determine the distribution of this ectoparasitosis in a rural community, an epidemiological study was carried out in a traditional fishing village in Ceara State, north eastern Brazil. Based on active case detection and voluntary participation, 91% of the population (belonging to 95% of all families) was examined. Embedded fleas were looked for over all parts of the body, counted, and the lesions were staged. The overall prevalence of infestation was 51.3% (95% CI: 47.0-55.5). More males than females were infested (54.8% vs 48.3%); however, this difference was not statistically significant. Age-specific prevalence rates followed an S-shaped curve with peaks in children aged 5-9 years and people elder than 60 years. The parasite burden was high (range 1-145 lesions; arithmetic mean: 8.9) and particularly elevated in males, children <15 years and the elderly. The distribution of the parasite burden was uneven within the population with the majority of the lesions in a few individuals: the 23 subjects (8% of all infested) with severe infestation (>30 lesions) accounted for 1,366 of the 2,493 lesions (54.8%) documented. The study shows that tungiasis is a highly prevalent ectoparasitosis in this deprived community with a peculiar distribution of prevalence and parasite burden. PMID- 12768414 TI - An Arabidopsis homologue of bacterial RecA that complements an E. coli recA deletion is targeted to plant mitochondria. AB - Homologous recombination results in the exchange and rearrangement of DNA, and thus generates genetic variation in living organisms. RecA is known to function in all bacteria as the central enzyme catalyzing strand transfer and has functional homologues in eukaryotes. Most of our knowledge of homologous recombination in eukaryotes is limited to processes in the nucleus. The mitochondrial genomes of higher plants contain repeated sequences that are known to undergo frequent rearrangements and recombination events. However, very little is known about the proteins involved or the biochemical mechanisms of DNA recombination in plant mitochondria. We provide here the first report of an Arabidopsis thaliana homologue of Escherichia coli RecA that is targeted to mitochondria. The mt recA gene has a putative mitochondrial presequence identified from the A. thaliana genome database. This nuclear gene encodes a predicted product that shows highest sequence homology to chloroplast RecA and RecA proteins from proteobacteria. When fused to the GFP coding sequence, the predicted presequence was able to target the fusion protein to isolated mitochondria but not to chloroplasts. The mitochondrion-specific localization of the mt recA gene product was confirmed by Western analysis using polyclonal antibodies raised against a synthetic peptide from a unique region of the mature mtRecA. The Arabidopsis mt recA gene partially complemented a recA deletion in E. coli, enhancing survival after exposure to DNA-damaging agents. These results suggest a possible role for mt recA in homologous recombination and/or repair in Arabidopsis mitochondria. PMID- 12768421 TI - Mandibular coronoid process in parathyroid hormone-related protein-deficient mice shows ectopic cartilage formation accompanied by abnormal bone modeling. AB - Parathyroid hormone-related protein (PTHrP) null mutant mice were analyzed to investigate an additional role for PTHrP in cell differentiation. We found ectopic cartilage formation in the mandibular coronoid process in newborn mice. While many previous studies involving PTHrP gene knockout mouse have shown that the cartilage in various regions becomes smaller, this is the first report showing an "increase" of cartilage volume. Investigations of mandibular growth using normal mice indicated that coronoid secondary cartilage never formed from E 15 to d 4, but small amount of cartilage temporally formed at d 7, and this also applies to PTHrP-wild type mice. Therefore, PTHrP deficiency consequently advanced the secondary cartilage formation, which is a novel role of PTHrP in chondrocyte differentiation. In situ hybridization of matrix proteins showed that this coronoid cartilage had characteristics of the lower hypertrophic cell zone usually present at the site of endochondral bone formation and/or "chondroid bone" occasionally found in distraction osteogenesis. In addition, the coronoid process in the PTHrP-deficient mouse also showed abnormal expansion of bone marrow and an increase in the number of multinucleated osteoclasts, an indication of abnormal bone modeling. These results indicate that PTHrP is involved in bone modeling as well as in chondrocyte differentiation. In situ hybridization of matrix protein mRNAs in the abnormal mandibular condylar cartilage revealed that this cartilage was proportionally smaller, supporting previous immunohistochemical results. PMID- 12768416 TI - Respiratory failure requiring extracorporeal membrane oxygenation after sodium phosphate enema intoxication. AB - A variety of adverse effects are associated with the use of hypertonic sodium phosphate enemas and laxatives in children. We describe an unusual case of phosphate enema toxicity in a child that resulted in cardiopulmonary failure necessitating the use of extracorporeal membrane oxygenation. The toxicity associated with hypertonic sodium phosphate enemas is reviewed. CONCLUSION: Hypertonic sodium phosphate enemas can cause cardiopulmonary failure. PMID- 12768422 TI - Three different fates of cells migrating from somites into the limb bud. AB - Cells from the ventrolateral dermomyotomal lips at limb levels undergo epithelio mesenchymal transition and migrate as individual and undifferentiated cells into the limb buds. The cells give rise to myocytes and blood vascular endothelial cells (BECs) in the limb. Using vascular endothelial growth factor receptor-3 (VEGFR-3) as a marker, it has also been shown that the somites contribute to endothelial cells of lymphatic vessels in the limbs, but it is unknown where the lymphangiogenic precursors are located within the somite. In this study we used the transcription factor Prox1 as a lymphatic marker and investigated whether cells in the dorso-lateral quarter of the somite differentiate into lymphatic endothelial cells (LECs) of the limbs. To label the migrating cells, the dorso lateral part of an epithelial brachial somite was grafted homotopically from quail into chick embryos at HH stages 13-14. The chick hosts were incubated until day 10-11 of development. The quail cell nuclei were identified with QCPN (anti quail) antibodies. Cell differentiation was analysed by immunohistochemical staining with QH1, anti-desmin and anti-Prox1 antibodies, and by in situ hybridisation with Prox1 probes. Our results confirm that quail cell nuclei are incorporated into the myotubes of the limb muscles. Quail cells are found in the endothelium of limb blood vessels and lymphatics, predominantly the dermal lymphatics. This indicates that superficial lymphatics develop independently from the deep ones and shows that cells migrating from the lateral somitic edge into the limb buds differentiate into three cell populations: myocytes, BECs and LECs. PMID- 12768423 TI - Preferential expression of the G90 gene in post-mitotic cells during mouse embryonic development. AB - G90 is a novel mouse gene that does not belong to any known gene family. It has previously been shown that this gene is expressed exclusively in post-mitotic cells of the adult mouse intestine and testis, therefore suggesting a role in the control of proliferation and/or differentiation. Here we report the detailed spatio-temporal expression pattern of G90 during mouse embryonic development. We found G90 expression in specific structures of the developing head, namely the brain, inner and middle ear, olfactory epithelium, vomeronasal organ, nasopharynx, oropharynx, papillae of the tongue and oral cavity, pituitary gland and epiglottis. Interestingly, there was a clear correlation between G90 expression and absence of proliferation in most of the cells showing expression of this gene during embryonic development; this finding supported our functional hypothesis. PMID- 12768424 TI - Comparative analysis of leg and antenna development in wild-type and homeotic Drosophila melanogaster. AB - The insect leg and antenna are thought to be homologous structures, evolved from a common ancestral appendage. The homeotic transformations of antenna to leg in Drosophila produced by mutation of the Hox gene Antennapedia are position specific, such that every particular antenna structure is transformed into a specific leg counterpart. This has been taken to suggest that the developmental programmes of these two appendages are still similar. In particular, the mechanisms for the specification of a cell's position within the appendage would be identical, only their interpretation would be different and subject to homeotic gene control. Here we explore the degree of conservation between the developmental programmes of leg and antenna in Drosophila and other dipterans, in wild-type and homeotic conditions. Most of the appendage pattern-forming genes are active in both appendages, and their expression domains are partially conserved. However, the regulatory relationships and interactions between these genes are different, and in fact cells change their expression while undergoing homeotic transformation. Thus, the positional information, and the mechanisms which generate it, are not strictly conserved between leg and antenna; and homeotic genes alter the establishment of positional clues, not only their interpretation. The partial conservation of pattern-forming genes in both appendages ensures a predictable re-specification of positional clues, producing the observed positional specificity of homeotic transformations. PMID- 12768425 TI - Seasonal variation in transcript accumulation in wood-forming tissues of maritime pine (Pinus pinaster Ait.) with emphasis on a cell wall glycine-rich protein. AB - Wood formation is being increasingly studied at cellular and biochemical levels; however, gene expression and regulation during wood formation remain poorly understood. Up to six types of wood can be studied within the same tree (early wood, late wood, juvenile wood, mature wood, reaction wood and opposite wood). These six types are characterized by different chemical, physical and anatomical properties. Using the cDNA-amplified fragment length polymorphism (AFLP) technique, we screened several thousand cDNA fragments from differentiating xylem of maritime pine (Pinus pinaster Ait.) comparing early wood vs. late wood and compression wood vs. opposite wood after 8 or 120 days of bending. About 100 transcript-derived fragments (TDFs) showed qualitative or quantitative variations between these different samples. The relative abundance of these TDFs was subsequently analyzed by reverse Northern using RNA derived from early and late wood. Analysis of variance (ANOVA) was used to identify differentially expressed TDFs ( P<0.01) and reverse transcription-polymerase chain reaction to confirm the differential expression of some TDFs. Among the genes with a known function, transcript expression and nucleotide sequence variation analysis showed a cell wall glycine-rich protein to be a strong candidate gene for wood properties. PMID- 12768426 TI - Relationship between shot put performance and triceps brachii fiber type composition and power production. AB - It is commonly accepted that shot put performance is mainly determined by the ability of the lower body to produce power. The purpose of the present study was to investigate the relationship between shot put performance and triceps brachii muscle fiber type composition and strength capacity. Thirteen male physical education students were selected to participate in the study based upon their shot put performance after 5 weeks of shot put technique instruction. At the completion of this technique-instruction period, they performed the following tests: shot put with a 6-kg shot, isokinetic torque measurements of the elbow extensors at 0, 0.52, 1.04, 1.57, 2.09, 3.14, and 4.19 rad.s(-1), maximal strength (1 RM) and explosive-throwing bench-press tests, one-arm seated shot put with 1-, 2-, 3-, 4-, 5- and 6-kg shot. Whole-body and dominant upper-arm bioimpedance measurements were used to estimate whole-body and upper-arm muscle mass. Muscle biopsy samples from the long head of the dominant triceps brachii were obtained and analyzed for fiber type composition with ATPase histochemistry. Shot put performance was significantly correlated with type II fiber area ( r=0.70, P<0.01), one-arm seated shot put (range r=0.60 to r=0.79, P<0.05), elbow extensors' isokinetic torque (range r=0.65 to r=0.78, P<0.05), bench-press tests ( r>0.86, P<0.01) and estimated arm muscle cross-sectional area ( r=0.68, P<0.05). These results suggest that fiber type composition and the functional capacity of triceps brachii muscle (e.g., isokinetic torque) explain a part of shot put performance. The magnitude of the correlation coefficients between shot put and the upper-body power tests suggests that other body parts (e.g., lower extremities) may play a significant role in this event. PMID- 12768427 TI - Problems with trials and intervention studies on barrier creams and emollients at the workplace. AB - The potential effect of barrier creams or emollients in the prevention of work related hand eczema has mostly been documented in a laboratory setting on experimentally damaged skin. Publications on real intervention studies of barrier creams or emollients in a workplace setting are scarce: only four such studies could be identified. The relatively short follow-up of most studies, and the small numbers of subjects enrolled, may reflect the organisational complexity of such studies. The observed effect is a combination of the intervention effect to be measured, and a number of disturbing variables. These disturbing variables are not just measurement error, but also phenomena such as the course of the disease, confounding and effect modification. In hand-eczema studies, the outcome parameters that should form the basis of the intervention effect are ill defined. Moreover, the study itself (i.e. the investigators) influences the results because there is rarely a single intervention: unknowingly, there are improvements in glove wearing, hand-washing habits or occupational hygiene in general. The performance of a blind study is often difficult or impossible. Large numbers of subjects have to be enrolled if a meaningful reduction in incidence of hand eczema is to be detected. One can demonstrate this by calculating the required enrollment in a study among employees in a nursing home. PMID- 12768428 TI - Skin cleansers for occupational use: testing the skin compatibility of different formulations. AB - OBJECTIVES: Skin-compatible hand cleansing is crucial for the prevention of occupational dermatitis. In this article we give an overview of skin-cleansing products and their ingredients and investigate whether solvents or abrasives could be avoided in cleansers for severely soiled skin. METHODS: Comparison of skin compatibility, cleansing power and effectiveness of different formulations of skin cleansers was performed via Duhring chamber tests [visual score, trans epidermal water loss (TEWL)] and standard hand-wash tests. RESULTS: The results show that, dependent on the grade of contamination, a range of skin cleansers that allow adequate skin cleansing, is necessary. Solvents could be avoided, by the use of powerful cleansing systems based on tenside/oil mixtures. Only for special contaminations may solvents be useful. Powerful cleansing activity could be performed without abrasives. CONCLUSIONS: The worker must be able to choose the skin cleanser that is adequate for the contamination that is present. Therefore, information and education of the workers are crucial for skin compatible skin cleansing. PMID- 12768432 TI - Use of PCR to detect infection of differentially susceptible maize cultivars using Ustilago maydis strains of variable virulence. AB - Ustilago maydis was specifically detected in infected maize plants by means of the polymerase chain reaction (PCR) using oligonucleotides corresponding to a specific region downstream of the homeodomain of the bE genes of the pathogen. The reaction gave rise to amplification of a ca. 500-bp product when tested with U. maydis DNA, but no amplification was detected with DNA from fungi not related to U. maydis. Using these primers, U. maydis was detected in infected maize plants from differentially susceptible cultivars as early as 4 days after inoculation with strains of variable degrees of virulence. Detection of U. maydis at early stages of infection, or in asymptomatic infected plants should assist in studies on plant-pathogen interactions. PMID- 12768431 TI - Unruptured large intracranial aneurysms in patients with transient cerebral ischemic episodes. AB - This report analyzes the treatment of six patients with transient cerebral ischemic episodes in combination with large unruptured aneurysm. Priority treatment of the symptomatic lesions had to be delayed in five cases because the large aneurysms were ipsilateral to stenoses of the internal carotid artery. The hemodynamic effect of surgery on the compromised cerebral circulation was pre examined with digital subtraction angiography and technetium-99m ((99m)Tc) single photon emission computed tomography. The data were correlated to aneurysm location and anatomical territory of the transient ischemic episodes. Only one patient showing a severe carotid stenosis contralateral to the large aneurysm, with decreased technetium uptake in the symptomatic hemisphere, required surgical correction of the stenosis first. In five patients, areas of reduced (99m)Tc uptake were mostly found within vascular boundary zones with angiographically verified effective collateral circulation. Clipping of the large aneurysms in these patients ipsilaterally to the stenotic lesion or on the anterior communicating artery as first option did not substantially increase the risk of subsequent stroke. PMID- 12768433 TI - Occurrence of MRSA endocarditis during linezolid treatment. PMID- 12768434 TI - Variants in the ALS2 gene are not associated with sporadic amyotrophic lateral sclerosis. PMID- 12768435 TI - Identification of mutations in the Bruton's tyrosine kinase gene, including a novel genomic rearrangements resulting in large deletion, in Korean X-linked agammaglobulinemia patients. AB - Mutations in the Bruton's tyrosine kinase ( BTK) gene are responsible for X linked agammaglobulinemia (XLA). We identified BTK mutations in six patients with presumed XLA from unrelated Korean families. Four out of six mutations were novel: two missense mutations (P565T, C154Y), a point mutation in a splicing donor site (IVS11+1G>A), and a large deletion (a 6.1-kb deletion including BTK exons 11-18). The large deletion, identified by long-distance PCR, revealed Alu Alu mediated recombination extended from an Alu sequence in intron 10 to another Alu sequence in intron 18, spanning a distance of 6.1 kb. The two known mutations consisted of one missense (G462D) mutation, and a point mutation in a splicing acceptor site (IVS7-9A>G). This study suggests that large genomic rearrangements involving Alu repeats are few but an important component of the spectrum of BTK mutations. PMID- 12768436 TI - Characterization of 458 single nucleotide polymorphisms of disease candidate genes in the Korean population. AB - Single nucleotide polymorphisms (SNPs) are considered as very promising genetic markers for complex disease gene hunting. However, it has been demonstrated that there are significant ethnic differences in genetic variations. In order to investigate the genetic variations in the Korean population and their ethnic differences, a large number of SNPs of 161 disease candidate genes were collected from a publicly available SNP database and then tested for the distribution of allele frequency in the Korean population. Of all 458 SNPs tested, approximately 43.9% were polymorphic in the Korean population, whereas 44.5% were monomorphic. The remaining 11.6% were failed in the test. Significant differences have been observed when SNP allele frequency pattern of Koreans was compared with those of Caucasians and Africans, whereas this pattern was highly similar between Korean and Japanese populations. Our data indicate that although many of the SNPs available in publicly available database, especially coding-region SNPs (cSNPs), can be used as informative genetic markers for disease association studies, an extensive verification of public SNPs in a particular population studied should be undertaken prior to their association studies. PMID- 12768438 TI - Identification of cartilage oligomeric matrix protein (COMP) gene mutations in patients with pseudoachondroplasia and multiple epiphyseal dysplasia. AB - Mutations in the cartilage oligomeric matrix protein (COMP) gene are responsible for two dominantly inherited skeletal dysplasias, pseudoachondroplasia (PSACH) and multiple epiphyseal dysplasia (MED). Mutation analysis of the COMP gene in Korean patients with PSACH and MED was performed. All nine patients with PSACH had mutations in the COMP gene, while three of the five patients with MED had detectable COMP mutations. Eight mutations, including three novel mutations, were identified in the COMP gene in the patients with PSACH and MED. Six mutations were found within the calmodulin-like repeats (CLRs) domain, especially in the seventh CLR and the other two mutations were in exon 16 outside of CLRs, which encode the C-terminal globular domain. Among the three novel mutations, two were missense mutations (Asp473Tyr, Asp482His) and one was a consecutive two-codon deletion, delAspAsp(469-473) in the five consecutive aspartic acid residues. All three novel mutations produced the PSACH phenotype. PMID- 12768437 TI - Molecular cloning and characterization of a novel human J-domain protein gene (HDJ3) from the fetal brain. AB - The J-domain is believed to be part of a chaperone involved in protein folding. From a fetal brain cDNA library, we isolated a cDNA of 3249 bp encoding a novel human J-domain protein, which was named as HDJ3. The expression pattern of HDJ3 was examined by reverse transcription/polymerase chain reaction, which suggested that the transcripts were highly expressed in human pancreas and selectively expressed in human brain, lung, liver, skeletal muscle and kidney. The results also showed that a probable splice variant of HDJ3 gene might exist. The HDJ3 gene was located on human chromosome 12q13.1-12q13.2 and consisted of seven exons spanning 8593 bp of the human genome. PSORT analysis indicated that the HDJ3 gene contained a transmembrane domain. The putative protein of the HDJ3 gene was highly homologous to rat dopamine-receptor-interacting protein, suggesting that it was a novel member of the molecular chaperone family and functionally related to dopamine signal transduction. PMID- 12768439 TI - Genetic variability of the extraneuronal monoamine transporter EMT (SLC22A3). AB - The extraneuronal monoamine transporter EMT (HGNC Nomenclature SLC22A3) is the molecular correlate of the classical uptake(2) system responsible for the non neuronal inactivation of circulating and centrally released catecholamines. Because of its functional profile and expression pattern, EMT is regarded as a candidate gene for diseases related to the sympathetic nervous system and neuropsychiatric disorders. We describe the first investigation of the genetic variability of the EMT gene in human. Six single-nucleotide substitutions and one deletion were detected within the assumed core promoter, the exonic and flanking intronic sequences and the 3'-untranslated region in 100 Caucasian individuals. No amino acid changes were found and Tajima's D was positive (D=2.91; P<0.01). However, the synonymous nucleotide substitution 1233G-->A might serve as a cryptic splice acceptor site. Analysis of linkage disequilibrium between polymorphisms yielded 12 possible haplotypes accounting for more than 90% of all haplotypes. Knowledge of the sequence variation and frequency of the underlying polymorphisms in this member of the amphiphilic solute facilitator family of transporters provides the basis for subsequent association studies and candidate gene approaches. PMID- 12768440 TI - The medaka fish Tol2 transposable element can undergo excision in human and mouse cells. AB - Tol2 is an active DNA-based transposable element identified in the medaka fish, Oryzias latipes. Originating from a vertebrate and belonging to the hAT ( hobo/ Activator/ Tam3) transposable element family, featuring a wide distribution among organisms, Tol2 would be expected to be active if introduced into mammals. We, therefore, examined if excision, one part of the transposition reaction, can occur in human and mouse culture cells. A Tol2 clone was introduced into cells and, after incubation, recovered. PCR and sequencing analysis provided evidence for precise and near precise excision in these cells. Tol2 can thus be expected to serve as a material for developing a gene transfer vector and other genetic tools applicable to mammals. It was also suggested that an intact Tol2 element could retain autonomy as a transposable element in mammalian cells. PMID- 12768441 TI - Association study of autoimmune thyroid disease at 5q23-q33 in Japanese patients. AB - As part of a genome scan to locate familial Graves' disease (GD) and Hashimoto's thyroiditis (HT) genes, an autoimmune thyroid disease (AITD) susceptibility locus has recently been identified at 5q31-q33 in a Japanese population. We performed an association study using six microsatellite markers located at this locus in a set of 440 unrelated Japanese AITD patients and 218 Japanese controls. We found significant allelic association between AITD and three markers located in 5q23 q33. GD demonstrated significant associations with two of these markers, while HT did not show significant associations with any markers. Further, when patients with GD were stratified according to clinical manifestations, the association was significantly different from the other subgroup of each category. These findings suggest the presence of susceptible genes of AITD, especially distinct subgroups of GD, in or near 5q23-q33. PMID- 12768443 TI - LMNA is mutated in Hutchinson-Gilford progeria (MIM 176670) but not in Wiedemann Rautenstrauch progeroid syndrome (MIM 264090). AB - Hutchinson-Gilford progeria syndrome (HGPS; MIM 176670) is an extremely rare disease that is characterized by accelerated aging and early death, frequently from coronary artery disease. Wiedemann-Rautenstrauch syndrome (WRS; MIM 264090) is another extremely rare disease that is characterized by progeroid features from birth with multiple somatic anomalies and paucity of subcutaneous fat. Because mutations in LMNA, encoding nuclear lamin A/C, cause other lipodystrophy syndromes, we sequenced LMNA (MIM 150330) from the genomic DNAs of seven unrelated HGPS probands and two unrelated WRS probands. We found four novel LMNA coding sequence variants among the HGPS probands, namely R471C, R527C, G608S and c.2036C>T. All seven HGPS probands had at least one LMNA variant, which were found in none of the genomes of 100 normal subjects ( P<4 x 10(-11)). In contrast, neither of the WRS proband genomes had any LMNA sequence abnormality. The strong association of rare LMNA coding sequence mutations with HGPS implicates this syndrome as a laminopathy, while WRS is most probably due to mutations in another gene. PMID- 12768442 TI - Association of interleukin-6 promoter variant with bone mineral density in pre menopausal women. AB - Interleukin-6 (IL6) has many roles essential to the regulation of the immune response, hematopoiesis, and bone resorption. Three single-nucleotide polymorphisms (SNP) in the IL6 promoter region were genotyped by the single-base extension method. The frequencies of each SNP were 0.002 ( IL6-597 G--> A), 0.27 ( IL6-572 G--> C), and 0.002 ( IL6-174 G--> C) in a Korean population ( n=1,082). IL6-597 G--> A and IL6-174 G--> C were totally linked together ( d(2)=1) and showed very low allele frequencies (0.002), which are common in Caucasians. On the other hand, the frequency of the IL6-572 G--> C*C allele was much higher (0.27) than that in Caucasian populations (<0.07). One of the IL6 promoter SNPs, viz., IL6-572 G--> C, showed significant associations with bone mineral density (BMD), i.e., the C allele was associated with increased BMD ( P=0.02, co-dominant model; P=0.007, dominant model). The mean BMD was highest in homozygous C individuals (0.67+/-0.15), lowest in homozygous G individuals (0.58+/-0.19), and intermediate in heterozygotes (0.64+/-0.21). In the present study, we describe a variant in the IL6 promoter region that shows positive association with higher BMD in a gene-dose-dependent manner in pre-menopausal women. PMID- 12768444 TI - Two cases of glucose-6-phosphate dehydrogenase-deficient Nepalese belonging to the G6PD Mediterranean-type, not India-Pakistan sub-type but Mediterranean-Middle East sub-type. AB - In Nepal, we tested 300 males for glucose-6-phosphate dehydrogenase (G6PD) activity. Two subjects were G6PD deficient (0.67%). Compared with normal controls, G6PD activity was 12% and 26%, respectively. The hemoglobin concentration of these two subjects was normal. We extracted genomic DNA from whole blood and read all sequences of G6PD. Both subjects had the same replacement of 563C>T, which was classified as G6PD Mediterranean. The amino acid might change from Ser to Phe at codon 188. These subjects also had a replacement of 1311C>T, which caused no replacement of an amino acid. A similar replacement pattern of G6PD Mediterranean is described from persons living in Mediterranean countries and Middle East countries. However, G6PD Mediterranean found in India and Pakistan has no replacement at nucleotide 1311. Thus, these two subjects in Kathmandu, Nepal, would be closer to people in Middle East countries than people in India. This is the first study of molecular analysis for G6PD deficiency in Nepal. PMID- 12768445 TI - Identification of novel polymorphisms in the Adam33 gene. AB - Adam33 is a member of a family of genes that encode membrane-anchored proteins with a disintegrin and a metalloprotease domain and that are primarily expressed in lung fibroblasts and bronchial smooth muscle. The human Adam33 gene is located on chromosome 20p13, a region that has been linked to asthma and bronchial hyperresponsiveness. Recently, the polymorphisms in Adam33 have been found to be associated with asthma. In this study, we performed polymorphism scanning of the entire genomic region, including the promoter region of Adam33, by direct sequencing. We identified 16 novel polymorphisms in the Adam33 gene. Among these novel polymorphisms, three polymorphisms (-2154G-->A, -753T-->A, and -330C-->T) were found to be in the promoter region and one polymorphism (13491 G-->A) was located in 3' untranslated region of the Adam33 gene. PMID- 12768446 TI - Quantitative structure-diastereoselectivity relationships for arylsulfoxide derivatives in radical chemistry. AB - Quantitative structure-diastereoselectivity relationships were studied for the intermolecular radical addition of deuterium and allyltributyltin to chiral arylsulfoxides by means of multiple linear regression and artificial neural networks (ANN). The values of diastereoselectivity (% syn) of the compounds studied were well correlated with the descriptors encoding the chemical structure. Using the pertinent descriptors revealed by the regression analysis, a square correlation coefficient of 0.9577 ( s=5.3825) for the training set was obtained for the ANN model in a 2-4-1 configuration. The results obtained from this study indicate that the diastereoselectivity of arylsulfoxide derivatives is strongly dependent on the shape of the R and X groups. FIGURE General structure of alpha-sulfinyl radicals PMID- 12768447 TI - Mutational analysis of Pyrococcus furiosus replication factor C based on the three-dimensional structure. AB - In eukaryotic DNA replication, replication factor C (RFC) acts as a "clamp loader" that loads PCNA onto a primed DNA template in an ATP-dependent manner. Proteins with functions essentially identical to that of RFC exist in Archaea. We have determined the crystal structure of the small subunit (RFCS) of Pyrococcus furiosus RFC at 2.8-A resolution. Using the information from the determined tertiary structure, we prepared several mutations in RFCS and biochemically characterized them. Truncation of the C-terminal alpha-helix (alpha16) causes a failure in RFCS oligomerization and a loss of the stimulating activity for the PCNA-dependent DNA synthesis by DNA polymerases. The site-directed reduction of the negative charges at the center part of the RFCS complex affected the stability of the RFC-PCNA interaction and reduced the clamp-loading activity. These results contribute to our general understanding of the structure-function relationship of the RFC molecule for the clamp-loading event. PMID- 12768449 TI - Cold-adapted yeasts as producers of cold-active polygalacturonases. AB - Eight cold-adapted, polygalacturonase-producing yeasts belonging to four species were isolated from frozen environmental samples in Iceland. They were identified as Cystofilobasidium lari-marini, Cystofilobasidium capitatum, Cryptococcus macerans and Cryptococcus aquaticus species by sequence analysis of rDNA regions. Growth behavior of the isolates was investigated. All strains could grow at 2 degrees C. Addition of glucose to pectin-containing culture medium had a repressive effect on enzyme production except for C. aquaticus, which showed increased polygalacturonase activity. Optimal temperature for enzyme production for the Cystofilobasidium strains was 14 degrees C, while that for the Cryptococcus strains was lower. Among the isolates, C. lari-marini S3B produced highest levels of enzyme activity at pH 3.2. Preliminary characterization of the polygalacturonases in the culture supernatant showed the enzyme from Cystofilobasidium strains to be optimally active at 40 degrees C and pH 5, and that from the Cryptococcus strains at 50 degrees C and pH 4. The polygalacturonase from C. macerans started to lose activity after 1 h of incubation at 40 degrees C, while that from the other strains had already lost activity at 30 degrees C. All the strains except C. aquaticus produced isoenzymes of polyglacturonase. In addition to polygalacturonase, the Cystofilobasidium strains produced pectin lyase, C. aquaticus pectin esterase, and C. macerans pectin lyase, pectate lyase and pectin esterase. PMID- 12768448 TI - Molecular identification of bacteria and Eukarya inhabiting an Antarctic cryoconite hole. AB - Inhabitants of a cryoconite hole formed in the Canada Glacier in the McMurdo Dry Valley region of Antarctica have been isolated and identified by small subunit (16S/18S) rDNA amplification, cloning, and sequencing. The sequences obtained revealed the presence of members of eight bacterial lineages (Acidobacterium, Actinobacteria, Cyanobacteria, Cytophagales, Gemmimonas, Planctomycetes, Proteobacteria, and Verrucomicrobia) and metazoan (nematode, tardigrade, and rotifer), truffle (Choiromyces), ciliate (Spathidium), and green algal (Pleurastrium) Eukarya. Bacterial recovery was approximately 20-fold higher at 4 degrees C and 15 degrees C than at 22 degrees C, and obligately psychrophilic bacteria were identified and isolated. Several of the rDNA molecules amplified from isolates and directly from cryoconite DNA preparations had sequences similar to rDNA molecules of species present in adjacent lake ice and microbial mat environments. This cryoconite hole community was therefore most likely seeded by particulates from these local environments. Cryoconite holes may serve as biological refuges that, on glacial melting, can repopulate the local environments. PMID- 12768450 TI - Growth physiology and competitive interaction of obligately chemolithoautotrophic, haloalkaliphilic, sulfur-oxidizing bacteria from soda lakes. AB - Two different groups of haloalkaliphilic, obligately autotrophic, sulfur oxidizing bacteria belonging to the genera Thioalkalimicrobium and Thioalkalivibrio have recently been discovered in highly alkaline and saline soda lakes. To understand response to their extreme environment and different occurrence in soda lakes, the growth kinetics and competitive behavior of several representatives have been characterized in detail using batch and pH-controlled continuous cultivation. The bacteria belong to the true alkaliphiles, growing within the pH range 7.5-10.6 with maximum growth rate and maximum growth yield at pH 9.5-10. On the basis of their response to salt content, three groups can be identified. All the Thioalkalimicrobium strains and some of the Thioalkalivibrio strains belonged to the moderate halophiles. Some of the Thioalkalivibrio strains from hypersaline soda lakes were extremely salt-tolerant and capable of growth in saturated soda brines. The Thioalkalimicrobium strains demonstrated relatively high specific growth rates, low growth yield, high maintenance, and extremely high rates of thiosulfate and sulfide oxidation. In contrast, the Thioalkalivibrio strains, in general, were slow-growing, high-yield organisms with lower maintenance and much lower rates of oxidation of sulfide and thiosulfate. Moreover, the latter survived starvation much better than Thioalkalimicrobium. Different growth characteristics and salt resistance appear to determine the outcome of the enrichment cultures from different soda lakes: Thioalkalimicrobium dominated in the enrichments with freshly obtained samples from diluted soda lakes at low-medium salinity, while Thioalkalivibrio was the predominant organism in enrichments from aged samples and at hypersaline conditions. In mixed thiosulfate-limited chemostat cultures at low salinity, Thioalkalimicrobium strains (mu(max)=0.33 h(-1)) out-competed Thioalkalivibrio strains (mu(max)=0.15 h(-1)) at D>0.02 h(-1). The overall results suggest that Thioalkalimicrobium and Thioalkalivibrio represent two different ecological strategies. PMID- 12768451 TI - Identification and characterization of the carAB genes responsible for encoding carbamoylphosphate synthetase in Halomonas eurihalina. AB - Halomonas eurihalina is a moderately halophilic bacterium which produces exopolysaccharides potentially of great use in many fields of industry and ecology. Strain F2-7 of H. eurihalina synthesizes an anionic exopolysaccharide known as polymer V2-7, which not only has emulsifying activity but also becomes viscous under acidic conditions, and therefore we consider it worthwhile making a detailed study of the genetics of this strain. By insertional mutagenesis using the mini-Tn 5 Km2 transposon we isolated and characterized a mutant strain, S36 K, which requires both arginine and uracil for growth and does not excrete EPS. S36 K carries a mutation within the carB gene that encodes the synthesis of the large subunit of the carbamoylphosphate synthetase enzyme, which in turn catalyzes the synthesis of carbamoylphosphate, an important precursor of arginine and pyrimidines. We describe here the cloning and characterization of the carAB genes, which encode carbamoylphosphate synthetase in Halomonas eurihalina, and discuss this enzyme's possible role in the pathways for the synthesis of exopolysaccharides in strain F2-7. PMID- 12768452 TI - Anoxynatronum sibiricum gen.nov., sp.nov. alkaliphilic saccharolytic anaerobe from cellulolytic community of Nizhnee Beloe (Transbaikal region). AB - New alkaliphilic anaerobic fermentative bacteria, strains Z-7981 and Z-7981', with Gram-positive cell walls, were isolated from the cellulolytic community from the soda lake Nizhnee Beloye, south-east of Baikal. Cells were motile rods, which differed in dimensions but, according to 98% DNA/DNA homology, belonged to the same species. Strain Z-7981 was chosen as the type and studied in detail. It did not produce spores and its cells were non-thermoresistant. It was a true alkaliphile with a growth range from pH 7.1 to pH 10.1 and optimal pH for growth at pH 9.1. It was obligately dependent on Na(+) and carbonate ions but not on Cl( ). Growth occurred in media with total sodium content from 0.076 M to 1.27 M Na(+ )with a broad optimum from 0.25 to 0.86 M Na(+). Growth showed an optimum at 35 degrees C, with absence of growth above 46 degrees C. The organism was aerotolerant and was capable of fermentation in non-reducing medium at less than 4.75% O(2) in the gas phase. Strain Z-7981 fermented mono- and disaccharides, sugar alcohols, but only glutamate and cysteine among the amino acids, and the proteinaceous substrates, chitin and dried Spirulina biomass. Fermentation products were acetate and ethanol. Fe(3+) was reduced in a process that yielded no energy. Phylogenetically the new organism belonged to cluster XI of the Gram positive bacteria with low G+C content and its closest neighboring taxon was Tindallia magadiensis. However, according to its phenotypic and genotypic characters it did not belong to any known genus from this group. We suggest a new genus and species with the name Anoxynatronum sibiricum and strain Z-7981 as its type (=DSM15060). PMID- 12768453 TI - Screening and characterization of the protease CP1 produced by the moderately halophilic bacterium Pseudoalteromonas sp. strain CP76. AB - A total of 26 proteolytic moderate halophiles were isolated and characterized. Most isolates were members of the genus Salinivibrio (16 strains), while others were identified as Bacillus (4 strains), Salinicoccus (2 strains), or members of the gamma-Proteobacteria (4 strains). Strain CP76 was selected as the best producer of an extracellular protease, designated CP1, and was used for further studies. Sequence analysis of the 16S rRNA gene in addition to phenotypic tests led to the placement of this organism in the genus Pseudoalteromonas. Maximal protease production was detected at the end of the exponential growth phase. This CP1 protease was purified and biochemically characterized, showing optimal activity at 55 degrees C, pH 8.5, and high tolerance to a wide range of NaCl concentrations (0 to 4 M NaCl). The most interesting features of this enzyme are its moderate thermoactivity, its activity at a range of pH values (6-10), and, especially, its salt tolerance (optimal activity at 7.5% total salts). The purified protease has a molecular mass of 38 kDa, and the N-terminal amino acid sequence determined showed similarity to metalloproteases previously described. The protease activity was strongly inhibited by EDTA, PMSF, and Pefabloc. No significant inhibition was detected with E-64, bestatin, chymostatin, or leupeptin. These results suggest that Pseudoalteromonas sp. strain CP76 produces an extracellular metalloprotease moderately thermotolerant and stable at high salt concentrations. PMID- 12768454 TI - Distribution of 16S rRNA introns among the family Thermoproteaceae and their evolutionary implications. AB - Novel 16S rRNA introns were detected in four new strains within the family Thermoproteaceae. Pyrobaculum oguniense TE7(T) and Thermoproteus sp. IC-062 housed introns of 32 and 665-668 bp after positions 1205 and 1213 ( Escherichia coli numbering system), respectively. Caldivirga maquilingensis IC-167(T) had two introns of 37 and 140 bp after positions 901 and 908, respectively. Vulcanisaeta distributa IC-065 had a 691-bp intron after position 1391. All the introns larger than 650 bp encoded the LAGLI-DADG type proteins. The intron-encoded proteins of P. oguniense TE7(T) and Thermoproteus sp. IC-062 are cognate with the proteins encoded by introns inserted at the same position in other Pyrobaculum/ Thermoproteus strains and phylotypes. The intron-encoded protein of V. distributa IC-065 is partially related to that of a Pyrobaculum phylotype. A large-scale deletion in the second intron of Caldivirga maquilingensis IC-167(T) is suspected. Based on these newly found introns and hitherto known 16S rRNA introns, the evolutionary movements of the 16S rRNA introns and the encoded LAGLI DADG type proteins are discussed. PMID- 12768456 TI - Tissue culture of the deep-sea eel Simenchelys parasiticus collected at 1,162 m. AB - We successfully cultivated fin cells of the deep-sea eel Simenchelys parasiticus (collected at 1,162 m) in L-15 medium supplemented with fetal bovine serum (FBS) and additional NaCl. We found that the pectoral fin cells proliferated in L-15 medium enriched with 4 g/l of NaCl salt (pH 7.3) containing 10% FBS at 10 degrees C and 15 degrees C. No cells were attached to the plastic culture plates when Dulbecco's modified Eagle's medium (pH 7.8) or 0-2 g/l of NaCl was added to the medium or when incubation was carried out at 4 degrees C. The majority of the explant outgrowth cells were detached when temperature increased to higher than 15 degrees C. The rate of proliferation of the fin cells was extremely slow and was dependent on the FBS concentration. Cell growth was enhanced by approximately 2.2-fold, and doubling time decreased from 170 h to 77 h when the FBS concentration was increased from 10% to 20% (v/v). Our established deep-sea eel cells were passaged 16 times over a 1-year period under atmospheric pressure conditions. PMID- 12768455 TI - Characterization of the precursor of tetraether lipid biosynthesis in the thermoacidophilic archaeon Thermoplasma acidophilum. AB - Polar lipid biosynthesis in the thermoacidophilic archaeon Thermoplasma acidophilum was analyzed using terbinafine, an inhibitor of tetraether lipid biosynthesis. Cells of T. acidophilum were labeled with [(14)C]mevalonic acid, and their lipids were extracted and analyzed by two-dimensional thin-layer chromatography. Lipids labeled with [(14)C]mevalonic acid, [(14)C]glycerol, and [(32)P]orthophosphoric acid were extracted and hydrolyzed under different conditions to determine the structure of polar lipids. The polar lipids were estimated to be archaetidylglycerol, glycerophosphatidylcaldarchaetidylglycerol, caldarchaetidylglycerol, and beta- l-gulopyranosylcaldarchaetidylglycerol, the main polar lipid of T. acidophilum. Pulse and chase experiments with terbinafine revealed that one tetraether lipid molecule is synthesized by head-to-head condensation of two molecules of archaetidylglycerol and that a sugar group of tetraether phosphoglycolipid is expected to attach to the tetraether lipid core after head-to-head condensation in T. acidophilum. A precursor accumulated in the presence of terbinafine with a fast-atom-bombardment mass spectrometry peak m/z 806 was compatible with archaetidylglycerol. The relative height of the peak m/z 806 decreased after removal of the inhibitor. The results suggest that most of the precursor, archaetidylglycerol, is in fully saturated form. PMID- 12768458 TI - Indications for and use of antidepressants in child and adolescent psychiatry--a cross-sectional survey in Denmark. AB - The prescription of antidepressants for children and adolescents is a controversial subject, and it has been documented that the practice has increased in the past decade in Denmark, the UK, and the USA. The aim of this study was to survey the indications for and use of antidepressants in child and adolescent psychiatry. Questionnaires were sent to all Danish child and adolescent psychiatric hospitals, out-patient clinics and privately practising psychiatrists treating children and adolescents under the age of 19 years (31 units in all). A 93.5 % response rate for the total of 382 questionnaires in the survey. The antidepressant serotonin selective re-uptake inhibitors (SSRIs) were the most prominently used agents in treating children and adolescents. The extent of their use represents 8 % of the total sample of individuals under the age of 19 years receiving any kind of psychiatric treatment - 0.03 % of the reference population in Denmark. It is only a surprisingly minor group of children and adolescents that are being treated with antidepressants despite the fact that 10 % of youth under the age of 19 are afflicted with diseases like depression, OCD, anxiety disorder and eating disorders. PMID- 12768457 TI - Isolation of a complete A1AO ATP synthase comprising nine subunits from the hyperthermophile Methanococcus jannaschii. AB - Archaeal A(1)A(O) ATP synthase/ATPase operons are highly conserved among species and comprise at least nine genes encoding structural proteins. However, all A(1)A(O) ATPase preparations reported to date contained only three to six subunits and, therefore, the study of this unique class of secondary energy converters is still in its infancy. To improve the quality of A(1)A(O) ATPase preparations, we chose the hyperthermophilic, methanogenic archaeon Methanococcus jannaschii as a model organism. Individual subunits of the A(1)A(O) ATPase from M. jannaschii were produced in E. coli, purified, and antibodies were raised. The antibodies enabled the development of a protocol ensuring purification of the entire nine-subunit A(1)A(O) ATPase. The ATPase was solubilized from membranes of M. jannaschii by Triton X-100 and purified to apparent homogeneity by sucrose density gradient centrifugation, ion exchange chromatography, and gel filtration. Electron micrographs revealed the A(1) and A(O) domains and the central stalk, but also additional masses which could represent a second stalk. Inhibitor studies were used to demonstrate that the A(1) and A(O) domains are functionally coupled. This is the first description of an A(1)A(O) ATPase preparation in which the two domains (A(1) and A(O)) are fully conserved and functionally coupled. PMID- 12768459 TI - Relationship between age of recognition of first disturbances and severity in young children with autism. AB - Autism is now thought to be present right from birth. Although usually not officially diagnosed until after the child's second birthday, parents often report disturbances before then. The age of detection of disturbances varies and may be linked to differences in the severity of the autism and its associated retardation. This study evaluates the developmental characteristics of 193 children with pervasive developmental disorder, using the same standard procedures for all subjects. Our goal was to determine the relationship between age of parental recognition of disturbances and disorder severity. The results indicated mainly a link between early abnormalities, associated medical condition and severity measured on cognitive tests. They suggest systematic screening for signs of autism in very young children. PMID- 12768460 TI - Perceptions of family relationships in adolescents with anorexia nervosa and their unaffected sisters. AB - The family relationships of patients with anorexia nervosa (AN) have been extensively studied over recent years. However, using case-control designs with unrelated controls is subject to various cultural and familial biases. Studying subjective differential perceptions of family relationships in sister-pairs discordant for the disorder may overcome some of these limitations. The aim of the present study was therefore, to investigate subjective perceptions of family environments in a clinically ill sample of female adolescent patients with acute AN and in their healthy sisters using the Subjective Family Image Test. We found significantly lower perceived individual autonomy and higher perceived cohesion in patients compared with their sisters but no difference in perceived emotional connectedness. Lower perceived individual autonomy of the ill children resulted mainly from their relationships with mothers but also in part from their relationships with fathers. This observed pattern might contribute to the maintenance of the disorder and should be addressed in individual and family interventions. PMID- 12768461 TI - Subscales scores of the Lithuanian version of CBCL--preliminary data on the emotional and behavioural problems in childhood and adolescence. AB - This study set out to investigate some psychometric properties of the Lithuanian version of CBCL for providing the preliminary data on the emotional and behavioural problems in Lithuanian children taking factors such as gender, age, SES and family composition into account. The CBCL was completed by parents of 7- to 11- and 12- to 14-year-old school children (N = 1296) drawn from the urban and suburban population. Younger boys scored higher than girls on externalizing problems and total problems scores, and older girls scored higher on internalizing problems. Younger children scored higher than the older children on total problems and externalizing problems scores. The comparisons with the US sample indicated that the levels of problem behaviour in 7- to 14-year-old children are similar to those found in the US. Lithuanian children had higher total problem scores; however, the effect sizes for other scales, except the attention problems scale and the somatic complaints scale, were very small. Similar to other studies, we found significant SES effects on total problem scores. Problem scores were highest for the lowest SES children. In general, this study supports previous findings concerning CBCL: similarities in emotional and behavioural problems outweigh differences in cross-cultural studies based on CBCL. PMID- 12768462 TI - Acamprosate and its efficacy in treating alcohol dependent adolescents. AB - BACKGROUND: About 50 % of adult alcoholic patients relapse within 3 months of treatment. Previous studies have suggested that acamprosate may help to prevent such relapse. The aim of our study was to assess the efficacy and safety of long term acamprosate treatment in alcohol dependence of adolescents. METHODS: In this, double-blind, placebo-controlled study, we recruited 26 patients, aged 16 19 years, with chronic or episodic alcohol dependence. Patients were randomly allocated treatment with acamprosate (1332 mg daily) or placebo for 90 days. Patients were assessed on the day treatment started and on days 30, and 90 by interview, self report, questionnaire, and laboratory screening. FINDINGS: 13 acamprosate-treated and 13 placebo-treated patients completed the treatment phase: of those withdrawn, 11 (1 vs 6) relapsed, 5 (3 vs 2) refused to continue treatment, 3 (1 vs 2) had concurrent illness, and 2 (1 vs 1) had adverse side effects. At the end of treatment, 7 acamprosate treated and 2 placebo-treated patients had been continuously abstinent (p = 0.0076). Mean cumulative abstinence duration was significantly greater in the acamprosate group than in the placebo group (79.8 [SD 37.5] vs 32.8 [19.0] days; p = 0.012). INTERPRETATION: Acamprosate is an effective and well-tolerated pharmacological adjunct to psychosocial treatment programmes. PMID- 12768463 TI - Longevity of direct resin composite restorations in posterior teeth. AB - This review is a survey of prospective studies on the clinical performance of posterior resin composites published between 1996 and 2002. Material, patient- and operator-specific data, observation periods, isolation methods of the operative field, and failure rates are detailed in tables. The data were evaluated statistically in order to assess the role of materials (filler size, bonding system, base materials [e.g. glass ionomer cements], and lining materials), study design, and personnel on failure rates. The primary reasons for composite failure were secondary caries, restoration fracture, and marginal defects. The influence of different commercial material brands on failure rates was not evaluated due to the great variety of test substances and the lack of material-specific documentation. Effects of the isolation method of the operative field (rubber dam or cotton rolls) and the professional status of operators (university or general dentist) on composite failure rates were not found to be significant. Observation periods varied from 1 to 17 years, and failure rates ranged between 0% and 45%. A linear correlation between failure rate and observation period was found (P<0.0001). Thirteen of 24 studies were terminated after 3 years, while seven studies continued for more than 10 years, indicating that favourable results for composite materials are frequently based on short term results, despite higher dropout rates in longer studies. To determine accurately the risk for patients, long-term, randomised, controlled clinical trials of treatment outcomes with composites used in posterior teeth are clearly needed. PMID- 12768464 TI - Dental treatment prior to stem cell transplantation and its influence on the posttransplantation outcome. AB - Patients who receive allogeneic stem cell transplantation (SCT) for hematological malignancies are at increased risk of developing oral complications. To reduce morbidity pretransplantation dental evaluation and treatment of all sources of potential infection have become standard of care for these patients. This study examined the effect of dental foci on the posttransplantation (post-SCT) outcome in two groups of patients who underwent allogeneic or autologous SCT: those who had no dental foci or completed dental treatment preoperatively (n=36) and those who underwent SCT without dental interventions (n=22). Statistical analysis showed no significant correlations between dental foci and infections, mucositis, and survival rate post-SCT. We therefore do not recommend a radical dental treatment pre-SCT. PMID- 12768465 TI - Stress fractures in athletes: review of 196 cases. AB - The purpose of this study was to investigate the association of stress fractures with age, sex, sport level, sporting activity, and skeletal site in athletes seen at our sports medicine clinic between September 1991 and May 2001. During these 10 years, 10 726 patients (6415 males, 3861 females) visited our clinic because of sport-related injuries, and 196 patients [125 males (1.9%), 71 females (1.8%)] sustained stress fractures. The average age of the patients with stress fractures was 20.1 years (range 10-46 years); 84 patients (42.6%) were 15-19 years of age, and 68 (34.7%) were 20-24 years of age. Altogether, 74 patients (37.8%) were active at the high recreational level and 122 (62.2%) at the competitive level. The sites of the stress fractures varied from sport to sport. The ulnar olecranon was the most common stress fracture site among baseball athletes and the rib among the rowing athletes. Classical ballet, aerobics, tennis, and volleyball athletes predominantly sustained stress fractures of the tibial shaft. Basketball athletes predominantly sustained stress fractures of the tibial shaft and medial malleolus and the metatarsal bone, whereas track and field and soccer athletes predominantly sustained stress fractures of the tibial shaft and pubic bone. Our results show that stress fractures are seen even in high-level adolescent athletes, with similar proportions for males and females, and that particular sports are associated with specific sites for stress fractures. PMID- 12768466 TI - Systematic review for evaluation of tolerability of nonsteroidal antiinflammatory drugs in osteoarthritis patients in Japan. AB - To evaluate the gastrointestinal tolerability of nonsteroidal antiinflammatory drugs (NSAIDs) in osteoarthritis patients in Japan, a systematic review of Japanese randomized controlled trials was performed. This study consisted of double-blind, randomized, controlled clinical trials with 4-week NSAID treatment of osteoarthritis patients in Japan. The analysis included 4725 patients from 25 trials. On average the cumulative incidences of patients who had experienced any adverse reaction and any adverse digestive reaction were 14.3% [95% confidence interval (CI) 13.3%-15.3%] and 10.4% (95% CI 9.4%-11.4%), respectively. The cumulative incidence for the upper gastrointestinal (GI) symptoms such as abdominal pain, nausea/vomiting, and dyspepsia was estimated to be approximately 10.9%. When the risk of upper GI symptoms was compared between males and females, the summary odds ratio was 1.71 (95% CI 1.11-2.65). Comparing the risk of upper GI symptoms between patients 59 years of age and younger and those 60+ years old, the summary odds ratio was 1.07 (95% CI 0.75-1.52). Despite the incidence of adverse reactions varying across the drugs being used, there was an obvious increased risk of GI symptoms. PMID- 12768467 TI - Validation of a Japanese patient-derived outcome scale for assessing total knee arthroplasty: comparison with Western Ontario and McMaster Universities osteoarthritis index (WOMAC). AB - We have developed a Japanese self-administered questionnaire based on an English version of the Western Ontario and McMaster Universities osteoarthritis index (WOMAC) to measure subjective function and pain status of patients who undergo a total knee arthroplasty procedure. Using multiple international cohorts, the performance of the developed Japanese scale was compared to the results of the WOMAC in the United Kingdom, United States, Canada, and Australia. The developed scale showed a comparable level of internal consistency and construct/criterion validity. The responsiveness of the scale was superior to the concurrently measured MOS Short Form 36 Physical Function scale. These results suggest that the developed scale is reliable, valid, and responsive for assessing the effectiveness of total knee arthroplasty in the Japanese context despite the cultural life style differences from Western countries. PMID- 12768468 TI - Fibroblast growth factor 2 in synovial fluid from an osteoarthritic knee with cartilage regeneration. AB - The levels of fibroblast growth factor 2 (FGF2) in synovial fluid of osteoarthritic knees were measured. The correlation between FGF2 and the severity of cartilage degeneration in varus-deformed knees with medial compartmental osteoarthritis or the articular cartilage regeneration that occurs after high tibial osteotomy (HTO) were investigated. Knees that underwent total knee arthroplasty (TKA) were categorized as either mildly or severely degenerated according to a modified Outerbridge's grading system for degeneration of articular cartilage. Regeneration of articular cartilage was observed in a biopsy specimen from the medial femoral condyle removed with the patient's consent during hardware removal approximately 2 years after HTO. The joint fluid FGF2 level was measured at that time using an enzyme-linked immunosorbent assay. Cartilage regeneration was classified as immature or mature according to the staging for regeneration of articular cartilage. The histological findings were analyzed using Pineda's evaluation method for cartilage regeneration. The mean concentration of FGF2 was 57.4 +/- 17.6 pg/ml in the joint fluid from knees with severely degenerated cartilage. This was higher than the FGF2 concentration found in the mildly degenerated group. Approximately 2 years after HTO the FGF2 level in synovial fluid was lower in knees with mature regenerated cartilage than in those with immature regeneration. Osteoarthritic knees at a more mature regeneration stage had a lower Pineda's histological score. This result suggested that the FGF2 concentration in knees with osteoarthritis was influenced by articular cartilage degeneration and regeneration, and it correlated with the histological evaluation. PMID- 12768469 TI - Basic and clinical studies of percutaneous radiofrequency ablation of osteoid osteoma using a standard electrosurgical generator. AB - For percutaneous radiofrequency ablation of osteoid osteoma for pain management, we used a standard electrosurgical generator instead of the radiofrequency generator system. First, we used the standard electrosurgical generator to determine the diameter of the coagulated area of normal femurs and humeruses of dogs under general anesthesia and to detect damage to normal tissue around the bone. We then used a standard electrosurgical generator to perform percutaneous radiofrequency ablation of the osteoid osteoma. All three patients were almost pain-free within the first 24 h after the procedure, and they were discharged the day after the operation, being hospitalized for only 2 days. We thus confirmed that percutaneous radiofrequency ablation using a standard electrosurgical generator produced results similar to those achieved with the radiofrequency generator system. PMID- 12768470 TI - Correction of tibial deformity in adults. AB - This report from five hospitals in Japan describes the results of correcting adult tibial deformities using external fixation. There were 49 patients with 59 lower limb deformities, with trauma being the most common cause of the deformity. Varus angulation was the most common deformity, and the most common magnitude was 11 degrees -30 degrees. Twenty-two patients had a leg-length discrepancy. The aim of the correction was to normalize both the mechanical axis and the inclination of the knee and ankle joints. In 63% of the patients corrections were performed gradually during bone lengthening or acutely after bone lengthening. Altogether, 71% of the patients were completely corrected, and no leg-length discrepancies remained after correction in 47%. Complications were encountered in 22 patients, about half of which were pin tract infections, 28% refractures, and the remainder delayed consolidation or fixator failure. There were no neurological or circulatory complications. The average fixation duration was 9 months. The average hospital charges were 3,740,000 yen in bilateral correction patients and 1,940,000 yen in unilateral correction patients. External fixation can correct not only the mechanical axis and joint inclination but also leg-length discrepancy simultaneously. PMID- 12768471 TI - Intramedullary nailing of multiple long-bone fractures of the lower extremity at the same surgery: a single-center experience. AB - We evaluated the outcome of multiple intramedullary nailing at the same surgery for traumatic long-bone fractures. All but 2 of 27 patients (average age 40) had been involved in motor vehicle accidents, and 17 of them presented with other associated nonorthopedic injuries. Ten patients had open fractures. Sixty-one nails were used. The average time from accident to fixation was 8.16 h, and the average time in the operating room was 5.4 h. No patients died; all required blood transfusion; and eight patients were admitted for at least 24 h to the intensive care unit. The average hospital stay was 27 days, the mean time to full weight-bearing was 4 months, and the average time to radiographic healing was 7.5 months. Three patients required dynamization to achieve fracture union; three underwent exchange nailing for nonunion and one for nail breakage. Thus, early fixation of multiple long-bone fractures by intramedullary nailing in polytrauma patients is an acceptable treatment method. PMID- 12768472 TI - Scaphotrapeziotrapezoid fusion: long-term follow-up study. AB - The long-term effectiveness of scaphotrapeziotrapezoid (STT) fusion is still controversial. We evaluated our clinical and radiological results of 30 STT fusions in 30 patients (average age 41 years; 23 males, 7 females). The follow-up period averaged 84 months. The indications were Kienbock's disease ( n= 23), isolated STT arthrosis ( n = 6), and dislocation of the trapezium ( n = 1). A total of 26 simultaneous associated procedures were performed. Excisional arthroplasty of the lunate was done in all 23 patients with Kienbock's disease. Of the 30 total patients, 5 had postoperative pain ranging from mild to severe; 4 patients were dissatisfied with the results. Postoperative ranges of motion of the wrist were 84% of extension and 91% of flexion compared with preoperative motion. The postoperative grip strength improved to 27 kgf from the preoperative value of 18 kgf. Altogether, 26 patients returned to their previous activities. All patients obtained uneventful STT fusion in an average of 11.2 weeks. Eight patients (27%) experienced postoperative complications; seven had radioscaphoid arthroses, five had trapeziometacarpal arthrodeses, and one had a flexor pollicis longus tendon rupture. Although radioscaphoid arthrosis occurred in 23% of this series after STT fusion, the results suggest that it is an effective procedure for Kienbock's disease and isolated STT arthrosis. PMID- 12768473 TI - Vascular anatomy for rotational acetabular osteotomy: cadaveric study. AB - Rotational acetabular osteotomies are performed to correct dysplastic hips in young adults. However, there is a potential risk of intrapelvic vascular injury. To define the relation of these vascular structures to the bone around the acetabulum, we measured the distance and direction from the anteroinferior iliac spine to the external iliac artery and from the base of the superior pubic ramus to the obturator artery in 34 cadaveric hemipelves (17 male, 17 female; 19 left, 15 right). The distance to the external iliac artery was significantly shorter in females (average 31.7 mm) than in males (average 38.2 mm); and the distance to the intrapelvic entry portal of the obturator canal, through which the obturator artery passes, was significantly shorter in females (average 27.2 mm) than in males (average 33.4 mm). In addition, the external iliac artery was located significantly more ventral and closer to the anteroinferior iliac spine in right hemipelves than in left hemipelves. The intrapelvic entry portal of the obturator canal was located more caudodorsal to the base of the superior pubic ramus in females than in males. Care should thus be taken during surgery in light of our findings. PMID- 12768474 TI - Genetic analysis of steroid-induced osteonecrosis of the femoral head. AB - With advanced organ transplantation technology, steroid-induced osteonecrosis of the femoral head (ONF) is one of the most troublesome complications. Steroid sensitivity varies among individuals, and the involvement of polymorphism of various genes relating to steroid metabolism is suggested. The present study investigated the relation between single nucleotide polymorphism (SNP) on the DNA sequence of cytochrome p(450) and ONF development. The subjects were 80 renal transplant patients. Genome DNA was obtained from the peripheral blood, and SNP analyses for CYP3A4, CYP2D6, and CYP2C19 were conducted using various methods: direct sequencing, polymerase chain reaction restriction fragment length polymorphism, and DNA Chip. The relation between ONF development and SNP was statistically analyzed. It is useful if the generating risk judged according to SNPs to prevention of steroid-induced ONF is possible. This time, SNPs, which are clearly related to ONF, were not accepted, although the possibility that SNP related to steroid metabolism is involved in ONF development is important. We think that it is necessary to examine this area more closely. PMID- 12768475 TI - Anteversion of the femoral neck in Indian dry femora. AB - The purpose of this study was to estimate the average angle of femoral neck anteversion in an Indian population. A total of 300 dry femora were classified by standard anatomical norms into male and female types and left-side and right-side types. They were evaluated by the Kingsley Olmsted and parallelograph methods, and the data were statistically analyzed. The average angles of anteversion obtained were 8.1 degrees and 8.3 degrees by the Kingsley Olmsted and parallelograph methods, respectively. The average female-type bone showed about 3 degrees more anteversion than the male-type bone. The average left-sided bone showed about 1.6 degrees more anteversion than the right-sided bone. A statistically significant difference existed between the sexes and the sides. The Kingsley Olmsted and parallelograph methods were found to be comparable with a correlation coefficient of 0.99. Altogether, 42.33%, 57.66%, and 79.00% of bones had readings of 5 degrees -10 degrees, 0 degrees -10 degrees, and 0 degrees -15 degrees, respectively. Thus, femoral neck anteversion has been found to be less in the Indian population than in Western populations. PMID- 12768477 TI - Synergistic induction of apoptosis of rheumatoid arthritis synovial cells by H(2)O(2) and N-acetyl-leucyl-leucyl-norleucinal. AB - The effects of proteolysis inhibitors on hydrogen peroxide (H(2)O(2))-induced apoptosis were examined in cultured human synovial cells of rheumatoid arthritis (RA) patients. RA synovial cells were resistant to apoptosis induced by H(2)O(2). In the presence of 100 microM N-acetyl-leucyl-leucyl-norleucinal (ALLN, known as calpain inhibitor 1 and also a proteasome inhibitor), but not N-acetyl-leucyl leucyl-methioninal (ALLM), apoptotic cell death was elicited by 400 microM H(2)O(2) at a concentration that alone never induced cell death. ALLN induced the expression of tumor suppressor p53 protein and p21(WAF-1) protein, probably through inhibition of proteasome. H(2)O(2) further potentiated ALLN-induced p53 expression. H(2)O(2) appeared to activate c-Jun N-terminal kinase (JNK) as well as extracellular signal-regulated kinase (ERK) and AKT. After administration of H(2)O(2) and p53 induction by ALLN, we found that either one alone is insufficient to induce apoptosis of RA synovial cells but their combination synergistically does so. These results suggest that induction of p53 by ALLN may be potentially important for triggering H(2)O(2)-induced apoptosis processes in RA synovial cells. PMID- 12768476 TI - Effect of dexamethasone on growth inhibition and chondrogenic maturation of human chondrosarcoma. AB - The effect of dexamethasone, a synthetic glucocorticoid, on in vitro and in vivo growth and differentiation of the human chondrosarcoma cell line (OUMS-27) was studied. Cells were treated with various doses of dexamethasone, and increasing doses produced an inhibitory effect on OUMS-27 tumor cell proliferation and induced maturation. Cell counts for OUMS-27 on day 9 ranged from 59% of the control at 10(-8) M to 45% of the control at 10(-5) M dexamethasone. Northern blot analysis revealed that the type II collagen mRNA level in cells given dexamethasone was lower than that in the controls, and the type X collagen mRNA level was higher than that in the controls. Phase-contrast microscopy revealed that cells grown in control medium formed monolayers consisting of small, polygonal cells, whereas dexamethasone-treated cells became larger and more irregular in shape. In the in vivo study the growth rate of masses in nude mice induced by inoculating OUMS-27 cells was also reduced in a dose-dependent manner with dexamethasone administration. These results suggest that dexamethasone caused growth inhibition and induced chondrogenic maturation of human chondrosarcoma cells. PMID- 12768478 TI - Comparison of the fit and fill between the Anatomic Hip femoral component and the VerSys Taper femoral component using virtual implantation on the ORTHODOC workstation. AB - The purpose of this study was to evaluate differences in fit and fill between an anatomic femoral component and a straight tapered femoral component, both of which were designed for proximal fit and fill using the preoperative planning workstation of the ROBODOC system (ORTHODOC). Anatomic Hip (Zimmer) and VerSys Taper (Zimmer) femoral components were each virtually implanted into 50 femora (25 dysplastic femora and 25 anatomically normal femora) using the ORTHODOC workstation. The fit and fill of the femoral components were measured on cross sectional images. The VerSys Taper femoral components showed significantly better fit and fill than the Anatomic Hip femoral components at the lower corner of the femoral neck cut and the middle of the femoral component in both the dysplastic femora and the anatomically normal femora. The Anatomic Hip femoral components showed significantly better fit and fill than the VerSys Taper femoral components 1 cm proximal from the femoral component tip in both dysplastic femora and anatomically normal femora. There was no significant difference in fit and fill between the two types of femoral component at the center of the lesser trochanter or 1 cm distal from the center of the lesser trochanter in either dysplastic femora or anatomically normal femora. Overall, VerSys Taper femoral components appear to provide better proximal fit and fill than Anatomic Hip femoral components in both dysplastic and anatomically normal femora. PMID- 12768479 TI - Difference of osteopontin gene regulation between bone and kidney. AB - Osteopontin is a sialoprotein that is expressed in various cells. It plays a variety of important roles in cell adhesion, migration, signaling, calcification, and immunity. Its diverse functions indicate that the regulation of osteopontin may also vary extensively among tissues. Although osteopontin promoter has been studied in vitro, in vivo analyses may be more appropriate for elucidating osteopontin's functions. In an attempt to investigate osteopontin gene expression, we generated transgenic mice in which the bacterial beta galactosidase reporter gene was conjugated downstream of osteopontin promoter. The osteopontin promoter was a mouse -910 bp upstream fragment, which we had previously found functional in 3T3 cells. Among 34 transgenic founders, 13 mice were transgenic, as determined with the polymerase chain reaction. Osteopontin and beta-galactosidase signals were evaluated with in situ hybridization. Among the 13 transgenic mice, 3 were beta-galactosidase-positive. In these transgenic mice, osteopontin signals were observed in bones and kidneys, whereas beta galactosidase message was detected only in bones. This suggests that the -910 bp osteopontin promoter is active in bones but not in kidneys. These data imply that the promoter region required for osteopontin expression in kidneys may differ from that in bones. PMID- 12768480 TI - Influence of spinal cord compression on the C3-C4 propriospinal neurons in cats. AB - The C3-C4 propriospinal neurons (PNs) transmit motor signals to forelimb motoneurons. Stem axons of the PNs bifurcate in the same segments and send ascending axons to the lateral reticular nucleus (LRN) as well as descending axons to the cervical motor nucleus. After laminectomy of decerebrated cats, ascending axons were stimulated by tungsten electrodes placed in the LRN. The N1 descending incoming volleys via axon reflexes with a latency of approximately 1.5 ms (N1 potential) were recorded in the forelimb motoneuron pools at C8 segment. We compressed the spinal cord at C3 or C6 segment by 6 x 5 mm plastic material and analyzed amplitudes of the N1 potentials at C8 motoneuron pools. After spinal compression step by step until 3000 microm from the surface for 30 min at the C3 segment, the amplitude of N1 potentials decreased to approximately 39.7% of the control. On the other hand, the amplitude of N1 potentials changed little after the same compression of the C6 segment. The N1 potentials during compression of the C3 segment did not recover to the initial amplitude of 63.5% after decompression. It is concluded that activities of axons at C3 segment are more sensitive than those at C6 to compression of the spinal cord. We discuss the properties of the bifurcation of the stem axons and the central spinal cord injury. PMID- 12768481 TI - Effects of liquid nitrogen treatment on the proliferation of osteosarcoma and the biomechanical properties of normal bone. AB - To overcome problems of autografts for reconstruction in the presence of malignant bone and soft tissue tumors, we devised a method for treating autografts that utilizes the hypothermic effect of liquid nitrogen. We measured temperature changes inside the bone at each condition and established a one-cycle liquid nitrogen protocol that included 20 min in liquid nitrogen, 15 min in room air, and 15 min in physiological saline. The proliferation potential of the tumor cells treated with the liquid nitrogen method was examined by means of bromodeoxyuridine (BrdU) immunostaining. Tumor proliferation potential in vivo was examined in nude mice. Based on the results we concluded that the tumor cells died out as a result of the liquid nitrogen method. Regarding compression strength there was no significant difference between intact bone and liquid nitrogen-treated bone, whereas the strength of the autoclaved bone was decreased. Scanning electron microscopic examination of the fracture surface of the autoclaved bone after the compression test showed an irregular, uneven surface, whereas that of the liquid nitrogen-treated bone was smooth and fine-grained. This might be one of the reasons for the discrepancy in compression strength. PMID- 12768482 TI - Correlation between femoral neck version and strain on the femur after insertion of femoral prosthesis. AB - Stress analysis on the femur after insertion of a femoral prosthesis with a modular interchangeable neck with adjustable neck version was performed using in vitro experimental procedures and cemented strain gauges. Although strain on the anterior and posterior aspects was less than on the medial and lateral aspects, it was more markedly affected by adjusting the interchangeable neck from straight to anteverted (or retroverted) than strain on the medial and lateral aspects, particularly around the stem tip. There was a highly significant correlation between compressive strain on the side toward which the prosthetic neck was oriented and the extent of neck version, whether anteverted or retroverted. Insertion of a femoral implant in excessive anteversion (or retroversion) may cause noticeably increased stress on the anterior and posterior aspects of the proximal femoral shaft. PMID- 12768483 TI - Induction of apoptosis of articular chondrocytes and suppression of articular cartilage proteoglycan synthesis by heat shock. AB - We investigated cellular and matrix responses of articular cartilage to heat shock. Rat articular cartilage was pretreated at 37 degrees C for 24 h before being exposed to 48 degrees C for 10 min and subsequently incubated at 37 degrees C for 1, 2, 4, 7, 10, and 14 days. Following heat shock, a terminal deoxynucleotidyl transferase nick end labeling assay showed that articular chondrocyte apoptosis appeared at day 1, peaked at day 7, and declined by day 14. Analysis by transmission electron microscopy confirmed that the chondrocytes had characteristic morphological features of apoptosis; immunohistochemical staining revealed that caspase-3 activity in chondrocytes increased, 3-B-3-positive articular chondrocytes decreased in number, and the expression of 3-B-3 native epitope in articular chondrocytes was reduced. Safranin-O staining revealed that depletion of proteoglycans in the matrix was not found in any group. Morphological and biochemical evidence from this study suggested that heat shock at 48 degrees C induced articular chondrocyte apoptosis and suppressed proteoglycan synthesis of articular cartilage in vitro. This study thus provides evidence of the onset of osteoarthritis induced by heat shock and a basis for choosing a temperature at which malignant bone tumor cells can be killed with minimal damage to articular cartilage. PMID- 12768484 TI - Effect of heat stimulation on viability and proteoglycan metabolism of cultured chondrocytes: preliminary report. AB - Thermotherapy has been applied to various joint diseases and injuries, but its direct effects on articular cartilage have remained unclear. The present study examined the effects on cell viability and metabolism by using the chondrocyte like cell line HCS-2/8. The temperatures and durations of heat stimulation were 39 degrees, 41 degrees, 43 degrees, and 45 degrees C for 15 or 30 min. After heat stimulation of 41 degrees C or lower for 15 or 30 min, cell viability increased and proteoglycan metabolism was accelerated, whereas after stimulation at 43 degrees C or higher for 30 min the viability and metabolism decreased. These results indicate that appropriate heat stimulation positively affects cell viability and the proteoglycan metabolism of articular cartilage, whereas too much heat stimulation produces negative effects. Clinical efficacy is therefore determined by the overall thermal dose. When the appropriate combination of temperature and duration is found, thermotherapy for diseases and injury of articular cartilage can be highly useful in clinical practice. PMID- 12768485 TI - Immunohistochemical study of the proliferation modality of synovium in rat adjuvant arthritis. AB - To investigate the proliferative potencies of the two types of synovial lining cells (types A and B), we used immunohistochemical techniques under light and electron microscopy to survey the complete process of arthritis in a rat model system. Complete Freund's adjuvant (0.1 ml) containing 5 mg of heat-inactivated Mycobacterium butyricum was administered intradermally into the right hind paws of 25 male Sprague-Dawley rats. Five animals were killed at weeks 1, 2, 3, and 4 after the immunological challenge, respectively, and the infrapatellar adipose synovium was removed from each animal. Using proliferative cell nuclear antigen (PCNA) as a cell proliferation marker, we quantified the locations of proliferative cells under light microscopy and then identified the cell type with immunoelectron microscopy. The number of PCNA-positive cells was high in the deep layer at week 2, and most of them were determined to be B cells. At week 3, the number of PCNA-positive cells increased in the superficial layer, and most were identified as A cells. Our results support the idea that cells of the synovial lining proliferate in situ during the course of arthritis. PMID- 12768486 TI - Enhancement of pedicle screw stability using calcium phosphate cement in osteoporotic vertebrae: in vivo biomechanical study. AB - We conducted an experimental study using female beagles with and without ovariectomy-induced osteoporosis to determine the effect of calcium phosphate cement (CPC) on the mechanical stability of inserted pedicle screws. A drill hole was created from the base of the transverse process to the vertebral body; CPC was injected into the hole, and then a screw was inserted into the same hole. In the presence of osteoporosis evidenced by dual X-ray absorptiometry, the stability of the inserted screw augmented by CPC against pull-out and cephalocaudal forces were significantly greater by 28% and 54% at 1 week after operation, 48% and 71% at 2 weeks, and 56% and 68% at 4 weeks compared with those without CPC. The pull-out strength increased progressively with time after surgery, probably reflecting new-bone growth from the surrounding cancellous bone, which was in direct contact with the CPC, as shown in the histologic study. At each time point the cephalocaudal rigidity was similar and the pull-out strength greater than that for the screws inserted without CPC in nonporotic dogs. These findings suggest that CPC augments the stability of the inserted pedicle screws and increases the stiffness of fixed osteoporotic motion segments using instrumentation. PMID- 12768487 TI - Effects of taurine administration in rat skeletal muscles on exercise. AB - To investigate the effects of taurine administration on exercise, we studied taurine concentrations in rat skeletal muscles after endurance running and the duration of running time to exhaustion, with and without taurine administration. For study 1 we divided 40 male SD rats into two groups: endurance exercise group ( n = 20) and sedentary control group ( n = 20). Each was further divided into two groups; one received distilled water ( n = 10) and the other taurine solution in water 0.5 g/kg/day orally ( n = 10) for 2 weeks. The exercise group performed treadmill running (60 min) once only after their nursing period. For study 2, we divided 10 male SD rats into two groups; one ( n = 5) received taurine 0.5 g/kg/day, and the other ( n = 5) received no taurine for 2 weeks; the two groups then performed treadmill running to exhaustion. In study 1, taurine administration increased taurine concentrations in leg skeletal muscles, whereas the concentrations were significantly lower in the exercised groups without taurine administration. Taurine administration reduced the decrease in taurine concentration in skeletal muscles on exercise. In study 2, the duration of running time to exhaustion was significantly increased by taurine administration. We concluded that peroral administration of taurine maintains the taurine concentration in skeletal muscle on exercise and up-regulates physical endurance. PMID- 12768488 TI - Salter Harris type II injury of the proximal phalanx of the fifth toe: case report. AB - Fractures of the growth plate are unique to childhood. The risk of growth disturbances depends on the type of fracture, its location, the age of the patient, the vascularization to the epiphysis, the state of the surrounding soft tissues, and whether the injury is open or closed. Epiphyseal fractures of the foot are uncommon, and only rarely are the phalanges involved. We report a Salter Harris type II injury in the fifth toe. The patient, a 3-year-old girl, was successfully managed conservatively with no sequelae. PMID- 12768489 TI - Combined fractures of the odontoid process and upper thoracic spine: two case reports. AB - We describe two cases of combined fractures of the odontoid process and upper thoracic spine that have not been previously reported. The first patient, a 21 year-old man, sustained an odontoid process fracture with posterior displacement and a fracture-dislocation injury at T4/5 in a motorcycle accident. The second patient, a 66-year-old woman, fell from a cliff and sustained an odontoid process fracture with posterior displacement and a bursting fracture at T3 and T4 with rotation. The first patient exhibited complete paraplegia below the T5 level of the spinal cord. The second patient escaped neurological deficit. Both underwent anterior screw fixation of the odontoid process and posterior fusion of the upper thoracic spine. In both cases the cervical spine seemed to be in hyperextension, and the upper thoracic spine experienced sudden flexion and rotation forces. PMID- 12768490 TI - Dumbbell-type solitary fibrous tumor in the cervical spine. AB - We report a dumbbell-type solitary fibrous tumor in the cervical spine. Eight spinal solitary fibrous tumors have been reported previously, but the cervical location is rare and this may be the first report of a dumbbell-type tumor. Histopathological examination showed that the tumor was composed of spindle cells in a collagen-rich matrix, although regional variability was noted. Diffuse immunostaining for CD34 and vimentin was noted in the cytoplasm of the tumor cells. This rare tumor should be recognized in the differential diagnosis of cervical spinal tumors. PMID- 12768491 TI - Mathematical model for the viscoelastic properties of dura mater. PMID- 12768492 TI - FDG-PET for evaluating musculoskeletal tumors: a review. AB - Positron-emission tomography (PET) can provide an in vivo method for evaluating metabolism and physiology in normal and diseased tissues. Clinical trials with [(18)F]2-deoxy-2-fluoro- d-glucose (FDG), the most commonly used radiolabeled tracer for PET imaging, have demonstrated increased accumulation of FDG in several cancer tissues. In this article, we introduce the basic principles of FDG PET and review current knowledge about FDG-PET for evaluating musculoskeletal tumors. Recent reports and our own experience suggest that FDG-PET cannot be a screening method for differential diagnosis between benign and malignant musculoskeletal lesions, including many neoplasms originating from different tissues altogether. FDG-PET might not accurately reflect the malignant potential of musculoskeletal tumors, but rather might implicate cellular components included in the lesions. A high accumulation of FDG can be observed in histiocytic, fibroblastic, and some neurogenic lesions, regardless of whether they are benign or malignant. More specific uses of FDG-PET, such as grading, staging, and monitoring of musculoskeletal sarcomas, should be considered for each tumor of a different histologic subtype. PMID- 12768493 TI - Computer-assisted orthopedic surgery. AB - Computer-assisted surgery (CAS) utilizing robotic or image-guided technologies has been introduced into various orthopedic fields. Navigation and robotic systems are the most advanced parts of CAS, and their range of functions and applications is increasing. Surgical navigation is a visualization system that gives positional information about surgical tools or implants relative to a target organ (bone) on a computer display. There are three types of surgical planning that involve navigation systems. One makes use of volumetric images, such as computed tomography, magnetic resonance imaging, or ultrasound echograms. Another makes use of intraoperative fluoroscopic images. The last type makes use of kinetic information about joints or morphometric information about the target bones obtained intraoperatively. Systems that involve these planning methods are called volumetric image-based navigation, fluoroscopic navigation, and imageless navigation, respectively. To overcome the inaccuracy of hand-controlled positioning of surgical tools, three robotic systems have been developed. One type directs a cutting guide block or a drilling guide sleeve, with surgeons sliding a bone saw or a drill bit through the guide instrument to execute a surgical action. Another type constrains the range of movement of a surgical tool held by a robot arm such as ACROBOT. The last type is an active system, such as ROBODOC or CASPAR, which directs a milling device automatically according to preoperative planning. These CAS systems, their potential, and their limitations are reviewed here. Future technologies and future directions of CAS that will help provide improved patient outcomes in a cost-effective manner are also discussed. PMID- 12768494 TI - Relevance of pathology and basic research to the surgeon treating rotator cuff disease. AB - For any physician expecting a successful outcome of a treatment regimen a thorough understanding of the underlying pathogenetic mechanism and the pathology of the disease process is an absolute prerequisite. In addition, the surgeon, obviously wishing to obtain a positive outcome of the procedure, must know the reaction of the body to his or her surgical actions. In particular, he or she must be familiar with the factors guaranteeing an uneventful healing process. For example, with rotator cuff disease it is important to realize that the site of degeneration leading eventually to tearing does not lie in the tendon itself but at its insertion into bone. Moreover, the cells and vessels needed for healing after surgical repair do not originate from the torn tendinous stumps. An important source of cells and vessels is the subacromial bursa overlying the site of tearing. Consequently, the bursa must be preserved at all cost. The subchondral bone trough into which the medial tendon stump is usually anchored during repair represents the other source of healing tissue. Whereas surgeons understandably concentrate their attention on the site of tearing, the fate of the muscle in the torn bone-tendon-muscle unit must not be neglected. In experimental studies we were able to measure muscle atrophy and fat accumulation and could quantify their evolution over time. Finding no reversal of these two parameters after successful repair was disturbing. Shoulder surgeons will benefit from this comprehensive review of updated concepts. PMID- 12768495 TI - Molecular cytogenetic analysis of solid tumors. AB - Molecular cytogenetic methods including fluorescence in situ hybridization (FISH) and comparative genomic hybridization (CGH) can be used for surgically removed solid tumors to obtain information valuable for both biomedical research and clinical oncology. FISH allows cytogenetic analysis even of cells in interphase. In addition, because CGH analysis permits comprehensive analysis of alterations in DNA copy number in a single experiment, it is possible to estimate not only the genetic pathways of carcinogenesis but also the biological characteristics, such as metastatic potential and patient prognosis at the time of diagnosing the solid tumor. The number of DNA copy number aberrations increases with tumor progression, leading to the concept of genetic staging of malignant tumors. Molecular cytogenetic analysis aids in realizing individualized, tailored medicine in cancer patients; therapeutic strategies are constructed for individual patients based on specific genetic alterations. PMID- 12768497 TI - Neuronal transmembrane chloride electrochemical gradient: a key player in GABA A receptor activation physiological effect. AB - It has long been accepted that GABA is the main inhibitory neurotransmitter in the mammalian brain, acting via GABA(A) or GABA(B) receptors. However, new evidences have shown that it may work as an excitatory transmitter, especially in the brain of newly-born animals and acting via GABA(A) receptors. The difference in the end results of GABA(A) receptors activation in the two cases is not due to the receptor associated channels, which in both cases are chloride channels. The different physiological effect in the two cases is due to different electrochemical gradients for chloride. When GABA acting via GABA(A) receptors is inhibitory, either there is no transmembrane electrochemical gradient for chloride or there is one forcing such negative ions into the nerve cell, once chloride channels are open. Viceversa, GABA is excitatory when the electrochemical gradient is such to make chloride ions flow outside the cell, upon opening of the GABA activated chloride channels.In this review this concept is discussed in details and evidence in the scientific literature for the existence of different types of chloride pumps (either internalizing or extruding chloride) is compiled. PMID- 12768498 TI - Regulation of renal amino acid (AA) transport by hormones, drugs and xenobiotics a review. AB - Major advances have recently been made in our understanding of the mechanisms and functions of amino acid transport in mammalian cells: - from the whole organism to transporter molecular structure. In this article, we present a brief overview of current knowledge concerning amino acid transporters, followed by a detailed discussion of the relevance of this new information to our broader understanding of the physiological regulation of amino acid handling in the kidney. We focus especially on the influence of hormones and xenobiotics on renal amino acid transport systems. In a growing number of cases, it now seems possible to correlate the effects of hormones, drugs, and xenobiotics with the capacity of renal amino acid transporters. This topic is of clinical relevance for the treatment of many amino acid reabsorption disorders. For example, under suitable conditions glucocorticoids and thyroid hormones stimulate renal reabsorption of amino acids and might therefore be of benefit in the treatment of different kinds of aminoaciduria. Hormonal regulation also underlies the postnatal development of renal amino acid reabsorption capacity, which can be stimulated to mature earlier after exogenous administration of e.g. glucocorticoids. In contrast, many compounds (e.g. heavy metal complexes) selectively damage renal amino acid transporters resulting in urinary amino acid loss. These types of phenomena (stimulation or inhibition of amino acid transporters in the kidney) are reviewed from the perspectives of our new molecular understanding of transport processes and of clinical relevance. PMID- 12768499 TI - The low nanomolar levels of N G-monomethylarginine in serum and urine of patients with chronic renal insufficiency are not significantly different from control levels. AB - There are no reliable mean values of N(G)-monomethylarginine (NMMA) in blood and urine of patients with renal insufficiency available in the literature. Therefore we investigate whether the NMMA levels are changed in blood and urinary excretion of nondialysed and dialysed patients with chronic renal insufficiency to evaluate whether NMMA may reach sufficiently increased concentrations in blood of the patients to exert toxic biological activity. In nondialysed as well as in dialysed patients we find no significant difference in serum concentration of NMMA between patients and controls. In nondialysed patients (all with a residual creatinine clearance lower than 15 ml/min), we find 94.5 +/- 26.1 nM (mean +/- SD) versus 94.6 +/- 19.5 nM in controls. Similar levels are found in serum of haemodialysed patients (each with serum creatinine levels >700 micro M): 83.0 +/- 20.2 nM. The urinary excretion of NMMA in nondialysed patients is also not significantly different from the excretion of controls: 123 +/- 110 in patients versus 157 +/- 117 nmol/24 hrs in controls. Furthermore, the clearance of NMMA is much lower compared to the clearance of the dimethylarginine derivatives. Based on the literature, the low nanomolar levels of NMMA found in blood of patients with renal insufficiency do not support the statement that NMMA proper may act as a uremic toxin. PMID- 12768500 TI - Effects of phenylalanine and phenylpyruvate on ATP-ADP hydrolysis by rat blood serum. AB - The nucleotide (ATP-ADP)/nucleoside (adenosine) ratio in the circulation can modulate the processes of vasoconstriction, vasodilatation and platelet aggregation. The main objective of the present study with rat blood serum was to evaluate the possibility of changes in nucleotide hydrolysis by phenylalanine (Phe) and phenylpyruvate (PP), the levels of which could increase in the circulation of individuals with phenylketonuria. Results demonstrated that Phe in the range 1.0-5.0 mM inhibited the ADP hydrolysis by rat serum. The effect of inhibition by Phe on ATP hydrolysis appeared only at a concentration of 5.0 mM. PP had no significant effect upon nucleotide hydrolysis. Kinetic analysis indicated that the inhibition of ADP and ATP hydrolysis by Phe in rat blood serum is uncompetitive. Conversely, Phe and PP did not affect the hydrolysis of p nitrophenyl-5'-TMP by rat serum. PMID- 12768501 TI - L-homoarginine suppresses exocrine pancreas in rats. AB - Previously, we found that guanidinated casein, a L-homoarginine-containing protein, was a more potent stimulator of pancreatic enzyme secretion than intact casein in rats. In this study, we examined secretory response and adaptation of the exocrine pancreas to the administration of free L-homoarginine in normal and bile-pancreatic juice (BPJ)-diverted rats. An intraperitoneal injection of L homoarginine (10 mg/rats) produced immediate and transient reduction in pancreatic secretion in BPJ-diverted rats, but not in normal rats. The BPJ diverted rats were fed with either a 25% casein, 45% casein, or 45% casein diet supplemented with L-homoarginine (19 g/kg diet) for 4 days. Feeding of a diet containing L-homoarginine inhibited the pancreatic adaptation induced by the high protein diet. These results indicate that L-homoarginine has an inhibitory effect on the secretion and production of exocrine pancreatic enzyme in BPJ-diverted rats, and L-homoarginine may have an antagonistic effect on CCK receptors. PMID- 12768502 TI - The role of ionotropic receptors of glutaminic acid in cardiovascular system. A. The influence of ionotropic receptor NMDA agonist - 1R,3R-ACPD and antagonist - DL-AP7 on the systemic pressure in rats. AB - The aim of our study was to estimate the involvement of the peripheral N-methyl-D aspartate receptors in regulation of cardiovascular function. For this purpose we examined the effects of intravenous injection of the agonists - NMDA (0.025; 0.05 and 1.0 mg/kg iv) and 1R-3R-ACPD (0.025; 0.05 and 1.0 mg/kg iv) - and antagonist of NMDA receptors DL-AP7 (0.02; 0.07 and 0.2 mg/kg iv). To determine if the effects of NMDA come from central or peripheral action we observed the effect during blockade of autonomic ganglion by using the nicotinic receptor antagonist chlorisondamine (1.25 mg/kg iv). Administration of NMDA in three doses evoked slight hypotension after injection of the medium dose, 0.05 mg/kg. In the condition of pretreatment with 1.25 mg/kg chlorisondamine the hypotensive effect of NMDA was markedly reduced, what might suggest that NMDA-induced hypotension raised from the action within the brain. The competetive NMDA receptor antagonist DL-AP7 slightly increased the blood pressure. None of the injected drug had an influence on the heart rate in our in vivo study. It is concluded that the peripherally localized NMDA receptors may take a part in regulation of cardiovascular system, since their stimulation or blockade evoked the changes of systemic pressure. PMID- 12768503 TI - Optimized conventional synthesis of "RGD" and "RGDS" peptides and their sarcosine mimics as integrin GP IIb/IIIa antagonists. AB - Synthetic arginyl-glycyl- alpha-aspartyl "RGD" and arginyl-glycyl- alpha-aspartyl serinyl "RGDS" peptide sequences, which are originally located in matrix proteins, are confirmed to be as versatile integrin GP IIb/IIIa antagonists. Since integrins, as cell surface glycoprotein receptors are implicated in several physiological mechanisms, these peptides are recently specially considered in the design of new therapeutics. Replacing glycine by sarcosine, as its more lipophilic isomer, in RGD peptides seemed, accordingly, interesting in revealing some structural/biological activity relationships. To render "RGD" peptides more conveniently available, an ameliorated quasi-gram yield conventional synthesis in solution of the parent "RGD" and "RGDS" [8, Scheme 1A & 15, Scheme 1B] and their sarcosine analogues, [8', Scheme 1A & 15', Scheme 1B] respectively, is herein described.A compilation of the mild hydrogenolysis removable Z and NO(2) groups and/or the acidiolytic removable Boc group were manipulated for the amino temporary protecting steps. Both the DCCI/HOBt and MA methodologies served well as peptide coupling methods. PMID- 12768504 TI - Serum amino acid concentrations in aging men and women. AB - The age and gender related differences in serum amino acid concentrations have been assessed in 72 (23-92 years) medically screened healthy men and women who were divided into three male and three female groups according to age. Free-time physical activity and food intake were analysed from the 5-day diaries. The subjects were instructed to eat according to their normal dietary habits and to avoid any clinical complementary nutritional products or other products that could increase protein or energy intake. The blood samples (5 ml) taken from the antecubital vein after an over-night fast were analysed for their amino acid contents by chromatography. In total nutrient intake of energy (P < 0.001), protein (P < 0.001), alcohol (P < 0.05), water (P < 0.01), sodium (P < 0.001) and fiber P < 0.001) decreased significantly with age. The concentration of total amino acids (P < 0.01), essential amino acids (P < 0.001), non-essential amino acids (P < 0.05) and branched-chain amino acids (P < 0.05) decreased, whereas citrulline (P < 0.001) and cysteine (P < 0.001) were the only amino acids, which increased with aging. In addition, men had significantly higher concentrations than women of essential amino acids (P < 0.001), branched-chain amino acids (P < 0.001), and 10 of the 22 individual amino acids assayed (P < 0.01). Women had significantly higher concentrations of aspartate (P < 0.05), glycine (P < 0.01), serine (P < 0.001) and taurine (P < 0.01) than men. It is concluded that the decrease in serum total amino acid concentration is associated with decreased energy and protein intake with aging and men have higher essential amino acid concentration in serum than women. PMID- 12768505 TI - Synthesis of new alpha-heterocyclic alpha-aminoesters. AB - alpha-Heterocyclic alpha-aminoesters were obtained in good yields by reaction of a glycine cation equivalent and different heterocyclic nucleophiles; diastereoselectivity using a carbohydrate (galactopyranose) as N-protecting group was modest. PMID- 12768507 TI - Metabolic interactions between restraint stress and L-lysine: the effect on urea cycle components. AB - We studied the effects of L-lysine on wrap-restraint stress-induced changes in ureagenesis. An exposure to wrap-restraint stress did not affect the plasma concentration of L-lysine, but did decrease plasma urea and arginine. Oral L lysine (1 g/kg) blocked the effect of stress on ureagenesis, and enhanced the effect of stress on L-arginine. No influence of L-lysine were found in controls. The results imply a stress-specific, ureagenesis-stimulating effect of L-lysine, and suggest an increased requirement for L-arginine during the above conditions. PMID- 12768506 TI - Phosphate-dependent glutaminase in enterocyte mitochondria and its regulation by ammonium and other ions. AB - The effects of ammonium and other ions on phosphate dependent glutaminase (PDG) activity in intact rat enterocyte mitochondria were investigated. Sulphate and bicarbonate activated the enzyme in absence and presence of added phosphate. In presence of 10 mM phosphate, ammonium at concentrations <1 mM inhibited the enzyme. This inhibition was reversed by increased concentration of phosphate or sulphate. The inhibition of PDG by ammonium in presence of 10 mM phosphate was biphasic with respect to glutamine concentration, its effect being through a lowering of V(max) at glutamine concentration of value of 0.01 was considered significant. Dropouts due to side effects were compared using the Fisher's exact X (2) test for two tails, a p>value of 0.05 was considered statistically significant. RESULTS: Propranolol as well as clonidine (p> = 0.005) proved to be statistically efficacious (p> = 0.0005). Propranolol was not more efficacious than clonidine (p> = 0.4). Dropouts were significantly higher in the clonidine group, 22 patients (p> = 0.006) and the most common undesirable side effect in this group was mouth dryness (in 8 patients). CONCLUSIONS: Clonidine was useful; when both drugs were compared propranolol was not shown to be superior to clonidine. Based on the results of this study, clonidine appears to be a good option for the treatment of mild to moderate essential tremor. PMID- 12768511 TI - [Memantine]. AB - Alzheimer's disease (AD) is the most common form of dementia in Western countries. The benefits presently observed with the approved treatments are mainly symptomatic without clear evidence of neuroprotection. N-methyl-D aspartate (NMDA) receptor antagonists have very extensive therapeutic potential in several central nervous system disorders and can be used as neuroprotective treatment in chronic neurodegenerative diseases and as symptomatic treatment in other neurologic diseases as epilepsy. Memantine, an antagonist of the glutamatergic NMDA receptor, has been recently approved for the treatment of advanced AD. Due to its action mechanism, memantine is considered a neuroprotective drug, whose utility has been demonstrated in preclinical studies, and a useful symptomatic treatment for AD and vascular dementia. We will review both aspects as well as the basic mechanisms mediating glutamatergic neurodegeneration and the implication of glutamate in cognition. PMID- 12768512 TI - [Extrapyramidal effects of antipsychotic drugs]. AB - Conventional antipsychotics have been used for many years to treat psychiatric and neurological disorders. Nevertheless, their effectiveness is often associated with a high incidence of side effects, especially extrapyramidal ones (EPS). With the entrance of the socalled atypical neuroleptics, this risk has been considerably, although not completely, reduced. Thus, clinicians should have good knowledge about the pharmacological characteristics of the newer anti-psychotics and their associated risk of worsening or developing motor disturbances. This article reviews these drugs, their pharmacological characteristics as well as the phenomenology of the extrapyramidal syndromes ans treatment of the EPS recommended. PMID- 12768513 TI - [Pneumoencephaly as a late complication of cranioencephalic trauma]. PMID- 12768514 TI - [Anterior opercular syndrome in childhood due to Moyamoya disease]. PMID- 12768515 TI - [Hemorrhagic stroke associated to neurocysticercosis]. AB - A well-known complication of neurocysticercosis is cerebral arteritis, which is usually manifested by cerebral ischemia. Only anecdotal cases of hemorrhagic stroke associated to this parasitosis have been described. Previously there are only two reported cases of this association. One of these cases had an intracystic hemorrhage confirmed by autopsy without cerebrovascular risk factors. Autopsy revealed an inflammatory arteriopathy adjacent to the cyst intracystic hemorrhage. The second case had a subarachnoidal hemorrhage secondary to the rupture of an aneurysm in the right anteroinferior cerebellar artery. At surgery, the aneurysm was found to be surrounded by a thickened-leptomeninges, which histologically showed the presence of cysticercous with dense inflammation. Our first patient was a 32 year-old female developed a lenticulo-capsular hemorrhage around a cysticercotic lesion. The second patient was a 34 year-old male developed an intracystic hemorrhage. As cerebral angiography was normal in both patients, cerebral hemorrhages were considered to be related to cysticercotic arteritis of small penetrating vessels. We conclude that cysticercosis is associated with differenttypes of intracranial hemorrhage, as documented the present cases. In neurocysticercosis endemic areas, cysticercotic arteritis should be added to the list of causes of intracranial hemorrhage in young people. PMID- 12768516 TI - [Importance of the clinical-radiological complementarity in superficial cerebral siderosis]. AB - We report two cases of superficial siderosis of central nervous system. In one case it was idiopathic and in the other secondary to an unidentified subarachnoid hemorrhage. The symptoms that characterized the clinical picture of both were gait disturbance and hypoacusis. The MRI study showed a superficial rim of hypointensity that covered the cerebellum and brainstem, and extended along the cranial nerves and the brain surface. The findings were clearer in the T2 spin echo series. Due to the high sensitivity for hemosiderin deposits, MRI made it possible to make the final diagnosis of this rare disease. PMID- 12768517 TI - [Treatment of sialorrhoea in neurological diseases with trans-dermic injections of botulinum toxin type A in the parotid glands]. AB - INTRODUCTION: Sialorrhoea is a disabling problem in many patients affected by neurological disorders. Anticholinergic drugs often produce side effects in these patients. Botulinum toxin A (TBA) blocks liberation of acetil-choline in motor and autonomic nerve terminals. METHODS: We report preliminary data about three patients, affected by motor neuron disease, Parkinson's disease and pons infarction respectively. All of them underwent treatment with TBA in both parotid glands to treat sialorrhoea, and were prospectively evaluated. Severity and Frequency Sialorrhoea Scale was used pre-treatment and at 6 weeks. TBA was administered in two points in each gland; a new injection was done at 2 weeks, with a dose ranging between 10 and 20 units in each point. RESULTS: Three males, with a men age of 63.6 years are reported. Sialorrhoea pre-treatment severity score was 5 while pre-treatment frequency score was 4 (continuous sialorrhoea all the days). Six weeks after TBA injection, two patients scored 2 in severity and frequency (mild sialorrhoea) and third patient had a score of 3 (moderate sialorrhoea). Side effects were observed neither after TBA injections nor at 6 week follow-up. CONCLUSIONS: Trans-dermic injection of TBA is safe and effective technique to treat sialorrhoea in neurological diseases when used by neurologists with experience, with few side effects. PMID- 12768518 TI - [Spontaneous spinal epidural hematoma]. PMID- 12768519 TI - ["Pseudo-stroke" hemiplegic type multiple sclerosis, a rare form of debut]. PMID- 12768520 TI - [Brave new world: cannabis for all? Some psychiatric remarks]. PMID- 12768521 TI - [Attention-deficit/hyperactivity disorder and substance abuse]. AB - Recent investigations of possible associations between attention deficit hyperactivity disorder (ADHD) and substance-use disorders are reviewed. ADHD seems to represent a possible risk factor for developing a substance abuse, up to 50% of patients with on continuing ADHD symptoms from childhood were found to develop a substance-use disorder. Based on possible pathophysiological similarities, especially cocaine and nicotine dependence are of focused interest in adolescents and adults with ADHD. The presence of ADHD may influence adolescent and adult substance-use disorders in different ways: earlier age of onset, higher frequency, longer duration of substance abuse and transition from alcohol-abuse to other substance-use disorders. Possible pharmacological and non pharmacological therapeutic strategies in patients with ADHD and substance abuse are discussed. PMID- 12768522 TI - [Emotional reactions of staff-members managing highly aggressive mentally ill patients]. AB - Highly aggressive behaviour of psychiatric in-patients often provokes intense emotional reactions and countertransference-feelings in staff especially when it is necessary to control aggression in psychiatric emergency units. We investigated the emotional reactions immediately after the manifestation of a patient's aggressive behaviour in a group of 53 staff members. The amount of aggression was measured with the help of the "Modified Overt Aggression Scale" (MOAS). The type and intensity of the staff-emotions can be adequately described by using a special self-rating questionnaire: "Eigenschaftsworterliste" (EWL 60 S). Using principal component analysis, three factors were differentiated: experience of stress, experience of sovereignty and experience of calmness. Several independent variables were identified as selectively increasing the probability of priority of each emotional factor mentioned above. In addition, a subgroup of staff members who have a raised risk of stress experience compared to the other staff members was compiled by cluster analysis. Based on an aggressiveness sequence model, useful strategies were discussed which directly aim to improve the coping abilities of staff members dealing with their emotional over involvement. PMID- 12768523 TI - [Comparison of ward atmosphere between three specialized wards for patients with alcoholism, schizophrenia, and depression and a diagnostic heterogeneous satellite ward]. AB - OBJECTIVE: Does the ward atmosphere differ between three specialized wards for patients with alcoholism, schizophrenia, and depression and a diagnostic heterogeneous satellite ward? METHODS: The ward atmosphere was assessed by patients and staff with the German version of the ward atmosphere scale (WAS). RESULTS: The data of 49 co-workers and 41 patients were evaluated. The satellite ward was rated significantly better than the specialized wards by both staff and patients (p < 0.01). CONCLUSIONS: If it is not possible to prove empirically, in future, that specialized wards have treatment advantages, our study suggests for atmospheric reasons that diagnostic heterogeneous wards should be recommended as standard care. PMID- 12768524 TI - [Semantic dementia--a variant of frontotemporal dementia]. AB - Relevant historical and clinical aspects of a case of semantic dementia are presented together with its neuropsychological and MRI findings. Differential diagnostic problems are studied in greater detail, in particular the differentiation to Alzheimer's disease. PMID- 12768525 TI - [The German version of the Community-Attitudes-Toward-the-Mentally-Ill (CAMI) inventory]. AB - OBJECTIVE: The German version of the Community Attitudes toward the Mentally Ill (CAMI) inventory developed by Taylor and Dear (1981) is presented. METHOD: The questionnaire was applied in a mail survey which was conducted in 3 small towns in Germany. RESULTS: At the item level surprisingly positive attitudes toward people with mental illness were found. The factor analysis with the 40 Likert scaled items yielded 4 dimensions (exclusion, integration, benevolence, social control) which account for 46 % of the total variance. As in previous studies, age had a negative effect and the level of education a positive effect on public attitudes, while the gender of the respondents had only a small effect. Familiarity with mental illness did not have an effect as important as in previous studies. CONCLUSION: There is a close correspondence between the German version and the original version of the CAMI-inventory. PMID- 12768526 TI - [The social situation of patients with late-life depression]. AB - The social situation of patients with late-onset late-life depression admitted to a gerontopsychiatric day-clinic was assessed with respect to activities, social contacts, living conditions and finances. The patients were compared with an age- and sex-matched control group. Relationships of the social situation to life satisfaction and severity of depression were investigated. In the patients, there were significant deficits in activities and contacts, especially with respect to those features which directly depend on the patients' level of activity (e. g. frequency of activities outside the household). Social infrastructure (e. g. frequency of attendance) and living conditions were positively correlated with life satisfaction, but were not significantly impaired in the depressed patients, neither were there differences in the financial situation between patients and controls. Thus, the social situation of elderly depressed patients is mainly characterised by a decreased level of activities, to which special therapeutic attention should be given. PMID- 12768527 TI - [Paraplegia after suicidal attempt]. AB - For the first time in Switzerland, patients were followed up who had become paraplegic after a suicidal attempt, and who had to spend a certain time in a center for paraplegics (corresponding to 3 % of all traumatically induced cases of myeloparalysis). Records and catamnestic data of patients with paraplegia after suicidal attempt during the years 1982 - 1996 (n = 38)--all being or having been patients of the centers for paraplegics f Basel and Nottwil (canton of Lucerne)--were evaluated. Catamnestic investigations performed from one month to 14 years after the suicidal attempt, based on a structured dialogue with a standardized, computerized questionnaire, led, among others, to the following conclusions: The average age of these patients (20 males and 18 females) was 38 years. The most frequently encountered suicidal method, leading to the paraplegic lesion, was a fall from a window of a building (89,6 %). In 55 % of the cases, psychiatric disease, especially depression, alcohol and drug dependence, had been diagnosed prior to the suicidal attempt. 37 % of patients hat attempted suicide at least once before the attempt leading to the paraplegic condition. 34 % had been hospitalized for psychiatric reasons on an earlier occasion. In 26 %, a farewell letter was written. As far as the level of the paraplegic lesions is concerned, patients were equally distributed (50 % each) between thoracal/cervical and lumbar lesions. The associated sexual impairment was experienced by each one of the 38 patients as a most disturbing problem. Despite the limited number of cases and the partially short time interval between the suicidal attempt and the follow-up investigation, results seem to indicate that such patients are not likely to commit suicide on a later occasion. One single patient had tried to commit suicide several times after the suicidal attempt event under discussion. PMID- 12768528 TI - [Pharmacological treatment of kleptomania--review]. AB - According to most diagnostic manuals such as ICD-10 and DSM-IV, kleptomania is classified as an impulse-control disorder. With regard to comorbidity, kleptomania is related to the obsessive- compulsive disorder spectrum and to the broader spectrum of affective disorders. Accordingly a psychopharmacological intervention with anti-depressant drugs or mood stabilizers may be possible, even though there are to date no known results from controlled therapy studies. Nevertheless the successful administration of such medication has been reported in several cases. Assuming a disturbed central serotonin reuptake, the use of selective serotonin reuptake inhibitors (SSRI) seems to be indicated. But also naltrexone could be of use in kleptomania. PMID- 12768529 TI - [Therapy resistant major depression: improvement of symptomatology after combining antidepressants with Tianeptine (Stablon)]. AB - OBJECTIVES: In common, depressive disorders are treated with various antidepressants, the number of which is enormously increasing. Nevertheless, in some cases therapy resistance is observed. In these cases, neuroleptics are used as an alternative. Recently, Tianeptin (Stablon) has also been used for the treatment of depressive disorders. METHOD: We report a 32-year old "therapy resistant" depressive female. Monotherapy with antidepressants and Tianeptin (Stablon) was ineffective. RESULTS: Finally, a combination of antidepressants and Tianeptin (Stablon) was effective and lowered the HAMD-Score significantly. DISCUSSION: A combination of antidepressants and Tianeptin (Stablon) seems to be an effective new Option for the treatment of "therapy resistant" major depressions. PMID- 12768530 TI - [Hawaiian baby rose wood: case report of an argyreia nervosa induced toxic psychosis]. AB - An increase of intoxications in persons using psychoactive plants and mushrooms can be observed. Mainly adolescents are experimenting with this group of plants for its hallucinogenic effects and easy and cheap availability. A case of a toxic psychosis with hallucinations, disturbances of orientation and psychomotoric agitation and anxiety after the intake of Hawaiian baby wood rose seed, an exotic ergotalkaloid containing plant often misused for its hallucinogenic effect is reported. Other drug-induced psychosis (e.g. LSD-psychosis), schizophrenic episodes or anticholinergic drug-induced intoxications have to be excluded. Regarding the increasing spreading of the misuse of psychoactive plants in Europe and North America intoxications with severe fatal sequelae have to be expected for the future. The knowledge of the clinical picture is important for correct diagnosis and treatment. PMID- 12768531 TI - Nonlinear synchronization in EEG and whole-head MEG recordings of healthy subjects. AB - According to Friston, brain dynamics can be modelled as a large ensemble of coupled nonlinear dynamical subsystems with unstable and transient dynamics. In the present study, two predictions from this model (the existence of nonlinear synchronization between macroscopic field potentials and itinerant nonlinear dynamics) were investigated. The dependence of nonlinearity on the method of measuring brain activity (EEG vs. MEG) was also investigated. Dataset I consisted of 10 MEG recordings in 10 healthy subjects. Dataset II consisted of simultaneously recorded MEG (126 channels) and EEG (19 channels) in 5 healthy subjects. Nonlinear coupling was assessed with the synchronization likelihood S and dynamic itinerancy with the synchronization entropy Hs. Significance was assessed with a bootstrap procedure ("surrogate data testing"), comparing S and Hs with their distribution under the null hypothesis of stationary, linear dynamics. Significant nonlinear synchronization was detected in 14 of 15 subjects. The nonlinear dynamics were associated with a high index of itinerant behaviour. Nonlinear interdependence was significantly more apparent in MEG data than EEG. Synchronous oscillations in MEG and EEG recordings contain a significant nonlinear component that exhibits characteristics of unstable and itinerant behaviour. These findings are in line with Friston's proposal that the brain can be conceived as a large ensemble of coupled nonlinear dynamical subsystems with labile and unstable dynamics. The spatial scale and physical properties of MEG acquisition may increase the sensitivity of the data to underlying nonlinear structure. PMID- 12768532 TI - Asymmetry of the brain surface from deformation field analysis. AB - The detection of asymmetry of exposed brain surfaces is examined, and a new method, deformation-based asymmetry (DBA), is introduced. DBA is based on analysis of two high-resolution magnetic resonance brain images, each with features representative of the subject group from which they were derived. Warping of individual brain images to their group representative image using octree spatial normalization provides sets of displacement vectors that are used in estimating deformation variance. For DBA group-representative left and right hemisphere images are compared. Representative hemisphere images are warped to each other and asymmetry analyzed using standardized d-values calculated as the ratio of displacement vector magnitude to the estimated component of variance in the direction of the displacement vector for each surface voxel. D-values were calculated within hemispheres by dividing subjects into two equal groups and comparing left-to-left and right-to-right. D-values from this ipsilateral hemisphere grouping were pooled. D-values from contralateral hemispheres were compared with the pooled ipsilateral hemisphere data. The proportion of d-values above a fixed level was used to test for difference between the two groups. High resolution magnetic resonance (MR) images from 20 young, right-handed males were studied using DBA. No significant differences were seen between sub-grouped ipsilateral d-values (P > 0.10). Highly significant asymmetries (P < 0.0001) were found between hemispheres, and in each lobe. Common right frontal and left occipital petalias were seen. The DBA method can theoretically be applied to any two groups of globally similar structures where analysis of dissimilarity of regional features is sought. PMID- 12768533 TI - More "mapping" in brain mapping: statistical comparison of effects. AB - The term "mapping" in the context of brain imaging conveys to most the concept of localization; that is, a brain map is meant to reveal a relationship between some condition or parameter and specific sites within the brain. However, in reality, conventional voxel-based maps of brain function, or for that matter of brain structure, are generally constructed using analyses that yield no basis for inferences regarding the spatial nonuniformity of the effects. In the normal analysis path for functional images, for example, there is nowhere a statistical comparison of the observed effect in any voxel relative to that in any other voxel. Under these circumstances, strictly speaking, the presence of significant activation serves as a legitimate basis only for inferences about the brain as a unit. In their discussion of results, investigators rarely are content to confirm the brain's role, and instead generally prefer to interpret the spatial patterns they have observed. Since "pattern" implies nonuniform effects over the map, this is equivalent to interpreting results without bothering to test their significance, a practice most of the experimentally-trained would eschew in other contexts. In this review, we appeal to investigators to adopt a new standard of data presentation that facilitates comparison of effects across the map. Evidence for sufficient effect size difference between the effects in structures of interest should be a prerequisite to the interpretation of spatial patterns of activation. PMID- 12768534 TI - A wavelet-based approximation of surface coil sensitivity profiles for correction of image intensity inhomogeneity and parallel imaging reconstruction. AB - We evaluate a wavelet-based algorithm to estimate the coil sensitivity modulation from surface coils. This information is used to improve the image homogeneity of magnetic resonance imaging when a surface coil is used for reception, and to increase image encoding speed by reconstructing images from under-sampled (aliased) acquisitions using parallel magnetic resonance imaging (MRI) methods for higher spatiotemporal image resolutions. The proposed algorithm estimates the spatial sensitivity profile of surface coils from the original anatomical images directly without using the body coil for additional reference scans or using coil position markers for electromagnetic model-based calculations. No prior knowledge about the anatomy is required for the application of the algorithm. The estimation of the coil sensitivity profile based on the wavelet transform of the original image data was found to provide a robust method for removing the slowly varying spatial sensitivity pattern of the surface coil image and recovering full FOV images from two-fold acceleration in 8-channel parallel MRI. The results, using bi-orthogonal Daubechies 97 wavelets and other members in this family, are evaluated for T1-weighted and T2-weighted brain imaging. PMID- 12768535 TI - Effects of age and speed of processing on rCBF correlates of syntactic processing in sentence comprehension. AB - Positron emission tomography (PET) was used to determine the effect of age on regional cerebral blood flow (rCBF) during syntactic processing in sentence comprehension. PET activity associated with making plausibility judgments about syntactically more complex subject object (SO) sentences (e.g., The juice that the child spilled stained the rug) was compared to that associated with making judgments about synonymous syntactically simpler object subject (OS) sentences (e.g., The child spilled the juice that stained the rug). In the first study, 13 elderly (70-80-year-old) subjects showed increased rCBF in the left inferior parietal lobe. This result contrasted with previous studies, which have shown activation in Broca's area in this task in young subjects. Elderly subjects were noted to have longer reaction times than young subjects previously tested. A second study found that young subjects whose reaction times were as long as those of the elderly subjects tested in Experiment 1 activated left superior parietal, and not left inferior frontal, structures. A third experiment found that elderly subjects with reaction times as fast as previously tested young subjects activated left inferior frontal structures. The results suggest that the speed of syntactic processing, but not age per se is related to the neural location where one aspect of syntactic processing is carried out. PMID- 12768537 TI - Adrenocortical zonation and ACTH. AB - The clear morphological distinction between the cells of the different adrenocortical zones has attracted speculation and experiment to interpret their functions and the ways in which they are regulated. Considerable data have been produced in recent years that has benefited a fuller understanding of the processes of steroidogenesis and of cell proliferation at the molecular level. This now enables the reexamination of earlier concepts. It is evident that there is considerable species variation, and this article, dealing mainly with the rat, reaches conclusions that do not necessarily apply to other mammals. In the rat adrenal, however, the evidence suggests that the greatest differences between the functions of the zones are between the glomerulosa and the fasciculata. Here the sometimes all-or-nothing demarcation in their complement of components associated with steroidogenesis or with cell proliferation suggests a stark division of labor. In this model the fasciculata is the main engine of steroid hormone output and the glomerulosa is the site of cell proliferation, recruitment, and differentiation. Regulating these functions are angiotensin II and other paracrine components that modulate and maintain the glomerulosa, and ACTH, that maintains the fasciculata, and recruits new fasciculata cells by transformation of proliferating glomerulosa cells. Grafted onto this mostly vegetative function of the glomerulosa is CYP11B2, limited to just a fraction of the outer glomerulosa in rats on a normal laboratory diet and generating aldosterone (and 18-hydroxycorticosterone) from precursors whose origin is not, from the evidence summarized here, very clear, but may include the fasciculata, directly or indirectly. The biosynthesis of aldosterone in the rat certainly requires reinterpretation. PMID- 12768538 TI - ACTH and adrenocortical gap junctions. AB - Since the initial identification of gap junctions in the adrenal gland, it has been proposed that a system involving direct cell-cell communication might be involved in adrenal cortical functions. Gap junction channels do, in fact, provide pathways for direct intercellular exchange of small molecules (<1,000 Da), many of which have the potential to influence a wide range of cellular activities. Gap junctions are composed of proteins called connexin which, in the adrenal cortex, have proven to be remarkably consistent in both type and zonal distribution with connexin 43 (Cx43) as the predominant component in mammalian adrenal glands thus far evaluated. Only the inner two zones of the cortex (zonae fasciculata and reticularis) exhibit significant amounts of Cx43 and functional coupling. Adrenocorticotropin (ACTH) has been shown to increase Cx43 protein in vivo and in vitro, and a strong correlation has been noted between the presence of gap junctions and certain adrenal cortical functions, especially steroidogenic capacity and cell proliferation. This review summarizes evidence of the Cx43 expression in adrenal cortical cells and the likely role of Cx43 in steroidogenesis and cell proliferation. It is concluded that control of gap junction expression in the adrenal gland is hormonally dependent and is functionally linked to adrenal gland zonation. PMID- 12768539 TI - Role of adrenocorticotropic hormone in the development and maintenance of the adrenal cortical vasculature. AB - The adrenal cortex is a highly vascularized endocrine tissue. A dense network of blood capillaries centripetally irrigates the adrenal gland, allowing every endocrine cell to be in contact with an endothelial cell. The pituitary hormone ACTH controls the coordinated development of the vasculature and the endocrine tissue mass. This suggests that paracrine secretions between steroidogenic adrenocytes and capillary endothelial cells participate in the control of adrenocortical homeostasis. Besides its effect on the vascular tone of arteries, ACTH induces the expression of the angiogenic cytokine VEGF-A (vascular endothelial growth factor-A) in primary cultures of adrenocortical cells. This growth factor is a specific mitogen for endothelial cells and is likely to mediate the hormonal control of adrenocortical vascularization through a paracrine mechanism. The newly discovered angiogenic factor EG-VEGF (endocrine gland-derived vascular endothelial growth factor), the expression of which is restricted to endocrine glands and which is preferentially mitogenic for endocrine tissue-derived endothelial cells, is another candidate mediator of great potential interest. PMID- 12768540 TI - Capsaicin-sensitive nerve fibers: a potential extra-ACTH mechanism participating in adrenal regeneration in rats. AB - Pituitary-derived factors, including ACTH, have been widely implicated in initiating adrenal regeneration. However, recent work has demonstrated that adrenal regeneration is also modulated by adrenal nerves that extensively reinnervate the regenerating adrenal. Moreover, transection of the splanchnic nerve removes sensory calcitonin gene-related peptide (CGRP) and preganglionic sympathetic vesicular acetylcholine transporter (VAChT)-positive fibers from the regenerating gland and delays regeneration. However, it is not known whether the splanchnic nerve effects on adrenal regeneration are mediated by the CGRP positive or VAChT-positive innervation. The present studies use the drug capsaicin, a neurotoxin selective for a subset of primary afferent neurons, to specifically remove CGRP-positive fibers from the adrenal gland and assess subsequent effects on the recovery of adrenal mass and function after surgical enucleation. Male, Sprague-Dawley rats were anesthetized and treated with capsaicin (vs. vehicle) periaxonally to the thoracic splanchnic nerve (33 mM, 15 minutes) or systemically (30-100 mg/kg for 4 days, s.c.). After 7-12 days of recovery, rats received right adrenalectomy and left adrenal enucleation. At 14 and 21 days postenucleation, prestress and poststress plasma and adrenals glands were collected; adrenals were weighed and fixed for immunolabeling of CGRP positive nerve fibers. Periaxonal capsaicin treatment decreased adrenal CGRP content prior to surgical enucleation; however, reinnervation by CGRP-positive fibers was not prevented and regeneration was not affected. Systemic capsaicin treatment attenuated the reinnervation by CGRP-positive fibers and increased the rate, but not extent, of adrenal regeneration. These results support the hypothesis that adrenal innervation represents an extra-ACTH mechanism to modulate the rate of adrenal regeneration. PMID- 12768541 TI - Adrenal neuropeptides: regulation and interaction with ACTH and other adrenal regulators. AB - It is now well accepted that both the cortex and medulla of the mammalian adrenal gland receive a rich innervation. Many different transmitter substances have been identified in nerves supplying both cortex and medulla and, as well as catecholamines, a wide range of neuropeptides has been found in the adrenal gland. There have been several studies on the affects of age, sodium intake, stress, ACTH, and splanchnic nerve activity on the regulation of adrenal neuropeptide content. There is evidence that the abundance of each of these peptides is actively regulated. Although there have been many studies addressing the individual actions of various neurotransmitters on steroid secretion, adrenal blood flow, and adrenal growth, few have attempted to determine the nature of any interaction between neurotransmitters and the classical adrenal stimulants. There are, however, some significant interactions, particularly in the regulation of zona glomerulosa function. This review necessarily focuses on vasoactive intestinal peptide (VIP) and neuropeptide Y (NPY), as these are the most abundant transmitter peptides in the adrenal gland and the majority of studies have investigated their regulation and actions. However, substance P, calcitonin gene related peptide (CGRP), neurotensin, and the enkephalins are included where appropriate. Finally, it has been suggested that certain neurotransmitters, particularly VIP, may interact with classical hormone receptors in the adrenal, notably the ACTH receptor. This review attempts to evaluate our current state of knowledge in each of these areas. PMID- 12768542 TI - Deconstructing the molecular mechanisms of cell cycle control in a mouse adrenocortical cell line: roles of ACTH. AB - This is a progress report of an attempt to deconstruct the signaling network underlying cell cycle control in the mouse Y1 adrenocortical cell line, aiming to uncover ACTH growth regulatory pathways. Y1 adrenocortical tumor cells possess amplified and overexpressed c-Ki-ras proto-oncogene. Despite this oncogenic lesion, Y1 cells retain tight regulatory mechanisms of cell cycle control typified by the sequential events comprising the mitogenic response triggered by FGF2 in G0/G1-arrested Y1 cells: 1) activation of ERK1/2 and PI3K, by 5 minutes; 2) induction of c-Fos and c-Myc proteins by 2 hours; 3) induction of cyclin D1 protein by 5 hours; 4) phosphorylation of Rb protein between 6 and 8 hours; 5) onset of DNA synthesis by 8-9 hours. In this cell line, ACTH-receptor (ACTH-R) activates contradictory pathways of growth regulation. First, ACTH coordinately induces fos and jun gene families via activation of both ERK1/2 and cAMP/PKA pathways, resembling a mitogen. Second, ACTH-R triggers cAMP/PKA-mediated antimitogenic mechanisms comprised of Akt/PKB dephosphorylation/deactivation, c Myc protein degradation, and p27(Kip1) protein induction. Induction of cyclin D1 depends on activation of both ERK1/2 and PI3K, but is not affected by ACTH action. As a consequence, ACTH antagonizes FGF2 mitogenic activity but ectopic expression of the c-Myc protein (via MycER fusion protein) is sufficient to abrogate this ACTH antagonistic effect over FGF2 mitogenic activity. Ectopic expression of both c-Myc and cyclin D1 is not sufficient to drive G0/G1-arrested Y1 cells into S phase, but when the sustained expression of these two proteins is complemented by ACTH treatment it promotes G1 phase progression and DNA synthesis initiation. In conclusion, ACTH-receptor lacks signaling potential sufficient to initiate a mitogenic response in Y1 adrenocortical cells and, therefore, cannot substitute for bona fide mitogens like FGF2. PMID- 12768543 TI - Mechanism of action of ACTH: beyond cAMP. AB - ACTH is the major regulator of adrenal cortex function, having acute and chronic effects on steroid synthesis and secretion. The precise molecular mechanisms by which ACTH stimulates steroid synthesis and secretion, as well as cell hypertrophy, survival, and migration are still poorly understood. Several studies have shown that ACTH action is mediated not only by cyclic adenosine monophosphate (cAMP), but also by calcium (Ca(2+)), both interacting closely through positive feedback loops to enhance steroid secretion. However, in spite of the evidence that ACTH could stimulate other signaling pathways, such as inositol phosphates and diacylglycerol or mitogenic-activated protein kinase pathway (MAPK), none is as potent as cAMP. Recent data indicate that duration and potency of the cAMP production could be modulated by several isoforms of adenylyl cyclases and phosphodiesterases. In addition, calcium is probably not a first second messenger per se; rather, there are several arguments indicating that its increase occurs following cAMP production. Finally, in addition to steroid secretion, ACTH, through cAMP, is a survival factor, protecting cells against apoptosis. All of the effects of ACTH are dependent on cytoskeleton integrity. In summary, after 30 years of intensive research in this field, cAMP remains the first obligatory second messenger of ACTH action. However, recent work emphasizes that cell environment (matrix and cytoskeleton) probably interacts with cAMP to coordinate functions other than steroid secretion. PMID- 12768544 TI - Adrenocorticotropin regulation of steroidogenic acute regulatory protein. AB - We studied the effect of the adrenocorticotropin hormone (ACTH) on the expression of the steroidogenic acute regulatory protein (StAR) in vivo in rat and hamster adrenals and also in transfection experiments using COS-1 cells. In vivo, ACTH increased the level of StAR mRNA within 30-60 minutes and also increased the quantity of StAR, but with a 2-3-hour delay. ACTH induced the formation of many acidic StAR species as analyzed by two-dimensional gel electrophoresis and immunoblotting. In the transfection experiments, (Bu)(2)-cAMP also induced the formation of many acidic species for the hamster StAR; in COS-1 cells, StAR is phosphorylated mainly on serine (S) residue(s). When alanine (A) was substituted for serine, S13A, S185A, and S194A mutants had decreased StAR activity compared to wildtype, thus determining the importance of these amino acid residues in StAR action. The full-length WT, N46-truncated StAR lacking its mitochondrial import sequence, and N46-S194A had similar activities, whereas N46-S185A had completely lost its activity. Our results suggest that S194, but not S185, functions in association with the mitochondrial import sequence for the initiation of StAR activation. Further studies showed that S185 is implicated in salt bridge stability, not in StAR phosphorylation, suggesting its importance for StAR folding. Thermodynamic calculations of the hamster StAR homology model based on MLN64 show that StAR can partially unfold to bind cholesterol and serve as a rapid transfer mechanism for eventual translocation into mitochondria. This is supportive of a StAR functioning either outside the mitochondria or in the mitochondrial intermembrane space. PMID- 12768545 TI - ACTH modulation of transcription factors responsible for steroid hydroxylase gene expression in the adrenal cortex. AB - Steroid hormone biosynthesis in the adrenal cortex and gonads involves the coordinated transcription of the genes encoding the steroid hydroxylases, 3beta hydroxysteroid dehydrogenase (3betaHSD), the steroidogenic acute regulatory protein (StAR), and adrenodoxin (Adx). Transcriptional regulation of steroidogenic genes is multifactorial, entailing developmental, tissue-specific, constitutive, and cAMP-dependent mechanisms. Optimal steroidogenic capacity is achieved by the actions of ACTH which exerts transcriptional pressure on all steroidogenic genes. The actions of ACTH in the adrenal cortex have been studied in great detail and is mediated by cAMP and protein kinase A (PKA) via two temporally distinct pathways. The acute response leads to mobilization of cholesterol, the initial substrate for all steroidogenic pathways, from cellular stores to the inner mitochondrial membrane where cholesterol sidechain cleavage cytochrome P450 (P45011A1) resides. The slower, chronic response of ACTH in the adrenal cortex directs transcription of the genes encoding the steroidogenic enzymes. Although steroidogenic gene transcription in response to ACTH is cAMP dependent, the consensus cAMP response pathway (CRE/CREB) is not involved. Instead, each steroidogenic gene utilizes unique cAMP-responsive sequences (CRS) found in the promoters of each gene, which bind a diverse array of transcription factors. Moreover, once specific transcription factors are bound to the promoters of the steroidogenic genes, increased gene expression requires posttranslational modification (phosphorylation/dephosphorylation) of the transcription factors and binding of coactivator proteins. This review provides a general view (with emphasis on the human) of the important factors involved in regulating steroidogenic gene expression and ultimately steroid hormone biosynthesis. PMID- 12768546 TI - Ectopic adrenocorticotropin (ACTH) and corticotropin-releasing hormone (CRH) production in the adrenal gland: basic and clinical aspects. AB - The hypothalamic-pituitary-adrenal (HPA) axis is integrated in the human stress system and controls the metabolism of many cell systems in the body. Therefore, hypofunction or hyperfunction of the HPA axis potentially threatens the life of the whole organism. Noncontrolled overproduction of its key regulators, CRH and ACTH, causes dysfunction of the stress system. Ectopic secretion of these compounds may be part of extraadrenal paraneoplastic syndromes caused by various benign or malignant tumors. However, ectopic ACTH and CRH may originate from the adrenal itself. A local CRH/ACTH system exists in the normal human adrenal medulla. Overproduction of CRH and ACTH has been documented in pheochromocytomas causing Cushing's syndrome. Finally, ectopic production of ACTH causing Cushing's syndrome has also been demonstrated in adrenocortical cells. This suggests a marked plasticity within the HPA axis and the neuroendocrine cell system. PMID- 12768547 TI - Immunohistochemical study of heat shock proteins 27, 60 and 70 in the normal human adrenal and in adrenal tumors with suppressed ACTH production. AB - Heat shock proteins (HSPs) are known to protect cells against various aggressions and to assist in the correct folding of nascent proteins as well as in the recovery of denatured ones. HSP70 increases its levels in the cell in response to any stress and is induced by ACTH in the adrenal gland. HSP60 is located in the mitochondria and assists in the folding of mitochondrial peptides. HSP27 is the only small HSP that is stress-induced. HSP27 and HSP70 are known to protect cells against apoptosis while, on the contrary, HSP60 is proapoptotic, increasing caspases maturation. We studied the expression of these HSPs in human adrenal tissue both in the normal glands (12 cases) and in tumoral tissue from cortisol producing adrenal adenomas (6 cases). Besides being neoplastic, these cells live in a particular ambience of lack of ACTH due to the suppression of the hypothalamic-pituitary ACTH secretion induced by the elevated levels of cortisol. HSP27 is highly expressed in the normal adrenal and shows a marked reduction of expression in Cushing's adrenal tissue. Although with overall lower levels of expression in the normal adrenal, HSP70 exhibited a similar pattern of reduction in tumoral tissue. HSP60, on the other hand, increased significantly and consistently in adrenal Cushing tumors. Besides the possible consequences of incorrect folding of nascent peptides, the alterations observed in tumoral tissue seem to act in an apoptotic direction. The only factor that we observed that could be contributing to these changes was the lack of plasma ACTH. PMID- 12768548 TI - Umbilical artery Doppler revisited: pathophysiology of changes in intrauterine growth restriction revealed. PMID- 12768549 TI - Color and pulsed Doppler ultrasonography of the fetal coronary arteries: has the time come for its clinical application? PMID- 12768550 TI - Coronary artery blood flow velocities in various fetal conditions. AB - OBJECTIVE: To determine fetal coronary artery peak blood flow velocities in normal and high-risk pregnancies. METHODS: Coronary artery peak systolic and diastolic blood flow velocities were measured by pulsed-wave Doppler velocimetry after identification of the coronary arteries by color Doppler imaging. Peak blood velocities obtained from normal pregnancies were related to gestational age using linear regression analysis. Blood flow velocities in normal fetuses were compared to measurements obtained in various fetal conditions. RESULTS: In normal fetuses coronary artery blood flow was visualized at a median gestational age of 33 + 6 weeks; median systolic and diastolic peak blood flow velocities were 0.21 and 0.43 m/s, respectively, and showed no significant change with gestational age. In growth-restricted fetuses coronary artery blood flow was visualized significantly earlier in gestation (median 28 + 2 weeks); systolic and diastolic peak blood flow velocities were higher at 0.25 and 0.48 m/s, respectively (P < 0.05). The highest coronary blood flow velocities were observed with fetal anemia. Coronary artery blood flow was also measured in fetuses with ductus arteriosus constriction due to indomethacin. Velocities did not differ from normal fetuses. Both in fetal anemia and ductus arteriosus constriction coronary artery blood flow could no longer be visualized with resolution of the underlying condition. CONCLUSION: Examination of coronary artery blood flow dynamics in the human fetus demonstrates acute increases in diastolic velocities in severe anemia and ductus arteriosus constriction based on the severity of the condition. In fetuses with growth restriction increased coronary blood flow velocities can be appreciated throughout the cardiac cycle. Clinical correlation in the interpretation of coronary blood flow dynamics in the human fetus is essential. PMID- 12768551 TI - Intrapartum assessment of fetal head engagement: comparison between transvaginal digital and transabdominal ultrasound determinations. AB - OBJECTIVE: To test the null hypothesis that no correlation exists between transvaginal digital examination and the gold standard technique of intrapartum transabdominal ultrasound determination of fetal head engagement, and secondarily to compare the performance of attending physicians and senior residents in depicting fetal head engagement by transvaginal digital examination. METHODS: Two hundred and twenty-two consecutive patients in labor > 37 weeks' gestation with normal singleton cephalic-presenting fetuses and with either ruptured or intact membranes were included. Of these, 119 were nulliparous and 103 were multiparous. Transvaginal digital examinations were performed by either attending physicians or senior residents and were followed immediately by transverse suprapubic sonographic assessments by a single sonographer. The fetal head was considered engaged on transvaginal digital examination if the leading part of the fetal head was positioned at least at maternal ischial spine station 0, and on transverse suprapubic ultrasound if the fetal biparietal diameter was below the maternal pelvic inlet. Examiners were blinded to each other's findings and the effect of examiner experience on the rate of agreement between the two techniques was assessed. The effect of several other independent variables upon the rate of agreement between the two modalities was also assessed. RESULTS: Overall, transvaginal digital examinations were consistent with ultrasound determinations with a raw percent agreement rate of 85.6% (95% confidence interval (CI), 80.8 90.3); kappa = 69.5% (95% CI, 59.4-73.9; P < 0.001). When stratified according to parity, the raw percent agreement rate for nulliparous patients was 81.5% (95% CI, 73.4-88.0); kappa = 60.7% (95% CI, 45.9-64.1; P < 0.001), and for multiparous patients it was 90.3% (95% CI, 84.1-95.9); kappa = 80.4% (95% CI, 63.0-87.5; P < 0.001). Maternal age, gravidity, maternal body mass index, gestational age, cervical dilatation, effacement, membrane status, ischial spine station of the fetal head, fetal head position at ultrasound assessment, birth weight and mode of delivery did not significantly affect rate of agreement. Parity did not affect examination consistency in multiparous patients, but in the whole group increasing parity increased the rate of agreement between modalities. Presence of combined spinal epidural anesthesia significantly increased the rate of agreement in the complete group and among multiparous but not among nulliparous patients. CONCLUSION: These data demonstrate a high rate of agreement (85.6%) between ultrasound determination and transvaginal digital assessment of fetal head engagement. Examiner experience had no effect. These data support the use of intrapartum transabdominal assessment of fetal head engagement. PMID- 12768552 TI - Comparison of transvaginal digital examination with intrapartum sonography to determine fetal head position before instrumental delivery. AB - OBJECTIVE: To investigate the accuracy of intrapartum transvaginal digital examination in defining the position of the fetal head before instrumental delivery. PATIENTS AND METHODS: In 64 singleton pregnancies undergoing instrumental delivery the fetal head position was determined by transvaginal digital examination by the attending obstetrician. Immediately after or before the clinical examination, the fetal head position was determined by transabdominal ultrasound by a trained sonographer who was not aware of the clinical findings. The digital examination was considered to be correct if the fetal head position was within +/- 45 degrees of the ultrasound finding. The accuracy of the digital examination was examined in relation to maternal and fetal characteristics. RESULTS: Digital examination failed to define the correct fetal head position in 17 (26.6%) cases. In 12 of 17 (70.6%) errors the difference was >/= 90 degrees and in five (29.4%) the difference was between 45 degrees and 90 degrees. The accuracy of vaginal digital examination was 83% for occiput-anterior and 54% for occiput-lateral + occiput-posterior positions. Logistic regression analysis demonstrated significant independent contributions in explaining the variance in the accuracy of vaginal examination for the station of the fetal head, the position of the fetal head and the experience of the examining obstetrician. CONCLUSIONS: Digital examination during instrumental delivery fails to identify the correct fetal head position in about one quarter of cases. PMID- 12768553 TI - Reference values for an index of fetal aortic isthmus blood flow during the second half of pregnancy. AB - OBJECTIVE: During fetal life, the parallel position of the two cardiac ventricles confers a special status to the aortic isthmus. Flow through the isthmus reflects the balance between the performances of the two ventricles and their respective peripheral impedances. This study proposes a fetal aortic isthmus flow velocity index and its reference values defined on the basis of gestational age (GA). METHODS: Video recordings of 111 normal fetuses from 18 to 39 weeks of gestation were retrospectively reviewed. An isthmus flow velocity index (IFI) was calculated as follows: IFI = (systolic + diastolic)/systolic velocity integrals. GA-specific reference ranges of IFI were constructed. RESULTS: An IFI of 1.33 +/- 0.03 was found at 18 weeks. This value decreased slightly but steadily with GA to reach 1.23 +/- 0.16 at 39 weeks. This change is mainly related to a decrease in diastolic velocity integrals. CONCLUSION: The proposed IFI provides information on the direction and, indirectly, on the volume of blood flow through the fetal aortic isthmus. PMID- 12768554 TI - Behavior of septum primum mobility in third-trimester fetuses with myocardial hypertrophy. AB - OBJECTIVES: The mobility of the septum primum (SP) in the fetus is a diastolic phenomenon and could be related to left atrial pressure. We studied the linear displacement of the SP in the left atrium in fetuses of diabetic mothers (FDM) with and without septal hypertrophy (SH) and in normal fetuses of normoglycemic mothers. In this study we set out to test the hypothesis that the linear displacement of the SP flap valve is less marked in fetuses with SH than in those without SH. METHODS: The ratio between the linear displacement of the flap valve and the left atrial diameter (excursion index (EI)) was compared in ten FDM with SH, eight FDM with normal septal thickness and eight normal fetuses of non diabetic mothers. Atrioventricular flow velocities were also compared in the three groups. RESULTS: Comparison of the three groups showed that in FDM with SH, the mean EI was 0.36 +/- 0.09, in FDM without SH it was 0.51 +/- 0.09 (P = 0.001) and in the control fetuses it was 0.49 +/- 0.12 (P = 0.03). There was a significant negative correlation between septal thickness and EI in FDM with SH. There was no correlation between septal thickness and atrioventricular flow velocities. CONCLUSION: Mobility of the SP in FDM with SH is reduced and there is an inverse correlation between the linear displacement of the SP and septal thickness. These findings may be related to changes in left ventricular diastolic function secondary to myocardial hypertrophy. PMID- 12768555 TI - The effect of metabolic control on fetal nuchal translucency in women with insulin-dependent diabetes: a preliminary study. AB - OBJECTIVE: Diabetes could potentially alter fetal nuchal translucency (NT) measurements mainly by increasing microvascular permeability. The aim of this study was to test this hypothesis. METHODS: Sixty-five women with pre-pregnancy insulin-dependent diabetes mellitus undergoing NT screening were evaluated. The main outcome measure was the NT measurement, expressed as both absolute measurements and delta values (number of standard deviations by which the observed value differed from the normal mean for the same gestation). The relationships between NT and years of diabetes, insulin dose, glycosylated hemoglobin, and glycemic capillary profiles ('tight' glycemic control) were studied. RESULTS: There was no significant correlation between NT and any of the studied parameters. There were no differences in NT according to the result of either glycosylated hemoglobin or 'tight' control. CONCLUSIONS: Metabolic control does not affect the result of ultrasound-determined fetal NT measurement which, when available, seems to be the best option for screening for Down syndrome in women with diabetes mellitus. PMID- 12768556 TI - Increased nuchal translucency and decreased fetomaternal transfusion after chorionic villus sampling. AB - OBJECTIVE: To investigate the relationship between nuchal translucency (NT) and fetomaternal transfusion (FMT) after chorionic villus sampling (CVS). METHODS: The level of FMT was determined in 272 viable, singleton pregnancies in which 10 14-week ultrasound scanning, NT measurement and CVS for fetal karyotyping had been performed. The pre-CVS NT was measured transvaginally, and the women were divided into two groups, i.e. those with NT < 2.5 mm (Group 1) or >or= 2.5 mm (Group 2). The level of FMT was determined via the maternal serum alpha fetoprotein levels before and after CVS. FMT was analyzed in relation to the pre CVS NT. RESULTS: Of the 272 pregnancies, 213 were in Group 1 and 59 in Group 2. The mean levels of FMT after CVS were 23.3 +/- 12.2 and 5.4 +/- 2.9 micro L in Groups 1 and 2, respectively (P < 0.01). An FMT > 100 micro L was found in 19 cases in Group 1, whereas the maximum in Group 2 was 67.2 micro L. Aneuploidies were diagnosed in 17 cases, 15 (88.2%) of them in Group 2. When the pregnancies with adverse outcome were excluded from the two groups, a higher level of FMT was observed in Subgroup 1 than in Subgroup 2 (P < 0.01). CONCLUSIONS: The mean level of FMT after CVS was significantly lower in pregnancies with an increased pre-CVS NT, a relationship observed in euploid pregnancies also. An increased pre-CVS NT seems to be inversely correlated with the FMT increase after CVS. Further studies are planned to investigate the background to this phenomenon. PMID- 12768557 TI - Clinical significance of normalization of uterine artery pulsatility index with maternal heart rate for the evaluation of uterine circulation in pregnancy induced hypertension. AB - OBJECTIVE: To investigate whether the use of normalized pulsatility index (PI) improves evaluation of the fetal prognosis in pregnancy-induced hypertension (PIH). METHOD: Eighty women with uncomplicated pregnancies and 46 women with PIH (33 with pre-eclampsia and 13 with gestational hypertension) were included in the study. Uterine artery PI and normalized PI were analyzed in relation to two neonatal parameters: incidence of small-for-gestational age infants and incidence of Cesarean section for non-reassuring fetal status. RESULTS: In normal pregnancy, the normalized uterine artery PIs (mean +/- SD) at 25, 31, and 39 weeks of gestation were 0.70 +/- 0.15, 0.71 +/- 0.16, and 0.65 +/- 0.13, respectively. Using non-normalized PI, 10 of 46 PIH cases had increased PI; however, with normalization, two pre-eclampsia cases were added to the group with elevated PI, giving a total of 12. The incidences of small-for-gestational age infants and Cesarean section due to non-reassuring fetal status were higher in the elevated normalized uterine artery PI group. The use of normalized PI appeared to reduce the false-negative results in pre-eclampsia and PIH. After normalization, sensitivity, positive predictive value and negative predictive value of PI for small-for-gestational age infants and Cesarean section due to non reassuring fetal status were improved in cases with pre-eclampsia and PIH. The incidence of these complications was low in gestational hypertension. CONCLUSIONS: Normalization of the PI may improve predictive values for small-for gestational age infants and delivery by cesarean section due to non-reassuring fetal status in PIH. PMID- 12768558 TI - Cervical length evaluation by transvaginal sonography in nongravid women with a history of preterm delivery. AB - OBJECTIVE: To evaluate cervical length in the nongravid period in women with a past history of preterm delivery. METHODS: The study population comprised 54 women who had delivered spontaneously before 34 weeks of gestation. Etiology of preterm delivery was suspected to be related to cervical incompetence, defined as a painless and progressive dilatation of the cervix in the absence of other causes. Patients with pre-eclampsia, intrauterine growth restriction, uterine anomalies, fetal anomalies, multiple gestation and similar complications were excluded. Shortening of the cervix was recorded during pregnancy in all study patients. One hundred and four women matched for age, parity and body mass index who had given birth at term served as the control group. Cervical length was evaluated in all women at least 12 weeks after delivery by transvaginal sonography in the mid-sagittal plane. RESULTS: The mean gestational age at delivery was 30.8 +/- 1.2 weeks in the study group and 38.8 +/- 2.1 weeks in the control group (P = 0.03). The mean cervical length in the two groups was 36 +/- 6 and 38 +/- 4 mm, respectively. This difference was not statistically significant (P = 0.9). CONCLUSIONS: In nongravid women with unexplained preterm delivery there is no difference in cervical length compared to patients who deliver at term. Shortening of the cervix is most probably a reversible phenomenon that occurs during pregnancy and represents a failure of the competence mechanism to adapt to pregnancy. PMID- 12768559 TI - Use of three-dimensional ultrasound imaging in the diagnosis of prenatal-onset skeletal dysplasias. AB - OBJECTIVE: Recognition of prenatal-onset skeletal dysplasias has improved with advances in ultrasound imaging. Skeletal abnormalities can be recognized by two dimensional (2D) ultrasound, but generating a precise diagnosis can be challenging. We aimed to determine whether three-dimensional (3D) imaging conferred any advantages over 2D imaging in these cases. METHODS: We studied five women with fetuses of 16-28 gestational weeks referred for abnormal ultrasound skeletal findings. First 2D and then 3D sonography was performed and the results compared. RESULTS: The pregnancies resulted in the following skeletal dysplasias: thanatophoric dysplasia, achondrogenesis II/hypochondrogenesis, achondroplasia, chondrodysplasia punctata (rhizomelic form) and Apert's syndrome. For all five fetuses, the correct diagnosis was made in the prenatal period by analysis of the 2D images. In each case the 3D images confirmed the preliminary diagnosis and for many findings it improved the visualization of the abnormalities. CONCLUSION: The 3D imaging had advantages over the 2D imaging when it came to evaluation of facial dysmorphism, relative proportion of the appendicular skeletal elements and the hands and feet. Most importantly, the patient and referring physician appreciated the 3D images of the abnormal findings more readily which aided in counseling and management of the pregnancy. PMID- 12768560 TI - Transrectal scanning: an alternative when transvaginal scanning is not feasible. AB - OBJECTIVE: In scanning the female pelvis the clear images of transvaginal sonography (TVS) result from placing the transducer close to the region of interest. The advantages of TVS over transabdominal sonography (TAS) and transperineal sonography are well documented. Transrectal scanning is proposed mostly for ultrasound guidance in draining a pelvic abscess. Our aim was to investigate the applicability of transrectal scanning (TRS) for cases in which TVS is impossible. METHODS: Forty-two patients with an absolute or a relative contraindication to TVS were scanned transabdominally and transrectally. The TRS was performed using a transvaginal probe, which was lubricated and slowly advanced into the rectum. The technique used was similar to that of TVS. Images were compared for resolution and quality. RESULTS: All scans were completed without significant patient discomfort or complaints. TRS was clearly superior to TAS in 31 cases. In nine cases TAS furnished some clinical information but TRS yielded better images. Only in one such case was TAS similar in quality to TRS. In four obese patients TAS did not reveal sufficient pelvic anatomy to generate a clinical diagnosis, whereas TRS revealed two sets of normal ovaries and two patients with ovarian cysts. In the two cases with vaginal agenesis TRS revealed the diagnosis of Rokitansky-Kuster syndrome. In three of the four patients with ruptured membranes the cervix could be measured precisely. CONCLUSION: Transrectal scanning should be used liberally after proper patient selection and counseling. The images obtained are superior to TAS and comparable to those obtained by TVS. PMID- 12768562 TI - Prenatal diagnosis of congenital leukemia in a fetus at 25 weeks' gestation with Down syndrome: case report and review of the literature. AB - Leukemia in a hydropic fetus with hepatosplenomegaly and Down syndrome was diagnosed at 25 weeks' gestation. Spontaneous demise occurred 10 days after the cordocentesis. A review of the literature of the seven cases described antenatally and 10 cases described in the immediate neonatal period or on examination of the stillborn baby is presented. PMID- 12768561 TI - Three-dimensional power Doppler imaging of ovarian stromal blood flow in women with endometriosis undergoing in vitro fertilization. AB - OBJECTIVE: The aims of this retrospective study were to investigate whether the quantification of ovarian stromal blood flow and/or leptin concentration are predictive of in vitro fertilization (IVF) outcomes in women after laparoscopic ovarian cystectomy for large endometriomas. METHODS: Twenty-two women undergoing IVF after laparoscopic surgery for ovarian endometriomas (> 6 cm) comprised the study group. Twenty-six women with tubal factor infertility constituted the control group. Ovarian stromal blood flow was evaluated by three-dimensional (3D) power Doppler ultrasound imaging using virtual organ computer-aided analysis (VOCAL( trade mark )). Serum and follicular fluid (FF) leptin concentrations were quantified using an enzyme-linked immunosorbent assay kit. RESULTS: There were significantly decreased ovarian stromal blood flow parameters (including vascularization index, flow index (FI), and vascularization flow index) in the endometriosis group without an evident difference in total ovarian volume on the day of human chorionic gonadotropin. The value of FF leptin demonstrated a negative correlation with ovarian stromal FI in the control group, but there was a loss of this effect in the endometriosis group. CONCLUSIONS: Quantification of ovarian stromal blood flow by 3D power Doppler ultrasound in women with endometriosis may provide an important prognostic indicator in those undergoing IVF. PMID- 12768563 TI - Prenatal diagnosis of hypoplastic left heart syndrome and trisomy 18 in a fetus with normal nuchal translucency and abnormal ductus venosus blood flow at 13 weeks of gestation. AB - We describe a case of early prenatal diagnosis of a major congenital heart anomaly and trisomy 18 in a low-risk pregnant woman. Nuchal translucency (NT) measurement at 13 weeks' gestation was 1.2 mm and Doppler evaluation of the ductus venosus detected a persistent reversed flow during atrial contraction. This finding prompted us to perform fetal echocardiography which showed hypoplastic left heart syndrome. Karyotyping following chorionic villus sampling diagnosed trisomy 18. Review of the recent literature suggests that the finding of an abnormal ductus venosus Doppler pattern in the late first trimester of pregnancy may be an early sign of either congenital cardiac or chromosomal abnormality, even in the presence of normal NT screening. PMID- 12768564 TI - Prenatal diagnosis of esophageal atresia with the pouch sign. AB - A 19-year-old primipara was referred to our center for a routine scan at 33 + 4 weeks' gestation. A visible but small stomach bubble (18 mm) was detected in the fetal abdomen, associated with a dilation of the esophagus with a tapering distal part. Direct visualization of filling and emptying of the proximal esophagus suggested the presence of obstruction. A cystic pouch in the region of the esophagus was observed to be full and empty in accordance with fetal swallowing. No other fetal anomalies were detected. Fetal biometric measurements were on the 10th percentile and polyhydramnios was detected. Esophageal atresia type I was suspected. Fetal karyotyping was declined by the parents, no therapy was given and the pregnancy continued until week 37 when a female baby was delivered following spontaneous labor. Esophageal atresia type I was radiographically confirmed. Bougienage was used for 8 weeks and esophageal anastomosis was successfully performed when the infant was 11 months old. No other anomalies were found and the child is currently in excellent health. The present case shows that the upper neck pouch sign may be a delayed manifestation present only in certain types of esophageal atresia. However, when it occurs it should prompt careful fetal examination as it is an important step in the diagnosis of esophageal atresia. PMID- 12768565 TI - New sonographic finding for the prenatal diagnosis of bladder exstrophy: a case report. AB - Bladder exstrophy is a very rare congenital malformation in which the anterior wall of the bladder is absent, and the posterior wall is exposed externally. Although sonographic findings of bladder exstrophy have been documented, we report a case presenting with a solid mass in the lower fetal abdomen with umbilical arteries running alongside the mass. The relationship between umbilical arteries and bladder exstrophy is a new sonographic finding which may be helpful in the prenatal diagnosis of this condition. PMID- 12768566 TI - Sonographic diagnosis of a uterine defect in a pregnancy at 6 weeks' gestation with a history of curettage. AB - We present the early diagnosis and successful surgical treatment of uterine perforation. This was a rare case of cystic change of a uterine perforation, which was diagnosed by sonography during the first trimester of pregnancy. Surgical closure of the uterine wall defect was successful. PMID- 12768567 TI - Placental site trophoblastic tumor masquerading as an ovarian ectopic pregnancy. AB - The vast majority of pregnancies of unknown location (PUL) will be failing pregnancies and early intrauterine or ectopic pregnancies (EPs) that are too early to visualize on transvaginal scan. Very rarely, a positive pregnancy test in the presence of a negative scan will reflect an underlying human chorionic gonadotropin (hCG)-secreting tumor. We report a case in which elevated serum hCG led to the initial diagnosis of an EP which was subsequently found to be a placental site tumor of the ovary. This case shows that a misinterpretation of signs of EP can result in a delay in accurate diagnosis. PMID- 12768568 TI - Maternal transcranial Doppler in pre-eclampsia and eclampsia. AB - Pre-eclampsia affects 3-7% of women and is associated with significant maternal and perinatal morbidity and mortality. Transcranial Doppler (TCD) has been used in pre-eclampsia/eclampsia to evaluate non-invasively the cerebrovascular hemodynamics in the maternal middle cerebral artery. TCD has demonstrated in pre eclamptic women maternal cerebral vasospasm, which does not correlate with mean arterial pressure assessed simultaneously. Estimated cerebral perfusion pressure, assessed using a modified formula, has been shown to be increased in women with severe and non-severe pre-eclampsia. However, in severe pre-eclampsia, elevated cerebral perfusion pressure is counterbalanced by increases in cerebrovascular resistance and cerebral blood flow is unaffected. In eclampsia a significant fall in cerebral vascular resistance occurs which, in the presence of increases in cerebral perfusion pressure, leads to hyperperfusion. Cerebral vascular changes to date have not been sensitive enough to predict the development of pre eclampsia or eclampsia. Longitudinal studies with the aim of predicting the onset of pre-eclampsia and to assess the effects of various drugs on the maternal cerebral circulation need to be designed. PMID- 12768569 TI - Light and scanning electron microscope study of nuchal translucency in a normal fetus. PMID- 12768571 TI - The mirror image artifact of early pregnancy. PMID- 12768572 TI - Use of color Doppler in the diagnosis of PPROM. PMID- 12768570 TI - Second-trimester sonographic demonstration of retrognathia and bilateral pyelectasis in a fetus with a duplication of chromosome 10q24.1-->qter. PMID- 12768573 TI - Changes in the proton T2 relaxation times of cerebral water and metabolites during forebrain ischemia in rat at 9.4 T. AB - Proton T(2) relaxation times of cerebral water and metabolites were measured before, during, and after transient forebrain ischemia in rat at 9.4 T using localized proton magnetic resonance spectroscopy ((1)H-MRS) with Hahn echoes formed at different echo times (TEs). It was found that the T(2) values of water and N-acetyl aspartate (NAA) methyl, but not total creatine (tCr) methyl, decrease significantly (approximately 10%) during ischemia, and this T(2) reduction is reversed by reperfusion. The T(2) reduction observed for NAA was most likely caused by the extravascular component of the blood oxygenation level dependent (BOLD) effect induced by a drastically increased deoxyhemoglobin content during ischemia. The absence of T(2) changes for tCr can probably be explained by the fact that the BOLD-related T(2) decrease was counterbalanced by the conversion of phosphocreatine (PCr) to creatine (Cr), which has a longer T(2) than PCr, during ischemia. The changes in T(2) should be taken into account for the quantification of metabolite concentrations during ischemia. PMID- 12768574 TI - Cluster analysis of BOLD fMRI time series in tumors to study the heterogeneity of hemodynamic response to treatment. AB - BOLD-contrast functional MRI (fMRI) has been used to assess the evolution of tumor oxygenation and blood flow after treatment. The aim of this study was to evaluate K-means-based cluster analysis as a exploratory, data-driven method. The advantage of this approach is that it can be used to extract information without the need for prior knowledge concerning the hemodynamic response function. Two data sets were acquired to illustrate different types of BOLD fMRI response inside tumors: the first set following a respiratory challenge with carbogen, and the second after pharmacological modulation of tumor blood flow using flunarizine. To improve the efficiency of the clustering, a power density spectrum analysis was first used to isolate voxels for which signal changes did not originate from noise or linear drift. The technique presented here can be used to assess hemodynamic response to treatment, and especially to display areas of the tumor with heterogeneous responses. PMID- 12768575 TI - Dynamic radial projection MRI of inhaled hyperpolarized 3He gas. AB - A radial projection sliding-window sequence has been developed for imaging the rapid flow of (3)He gas in human lungs. The short echo time (TE) of the radial sequence lends itself to fast repetition times, and thus allows a rapid update in the image when it is reconstructed with a sliding window. Oversampling in the radial direction combined with angular undersampling can further reduce the time needed to acquire a complete image data set, without significantly compromising spatial resolution. Controlled flow phantom experiments using hyperpolarized (3)He gas exemplify the temporal resolution of the method. In vivo studies on three healthy volunteers, one patient with chronic obstructive pulmonary disease (COPD), and one patient with hemiparalysis of the right diaphragm demonstrate that it is possible to accurately resolve the passage of gas down the trachea and bronchi and into the peripheral lung. PMID- 12768576 TI - Sensitive CEST agents based on nucleic acid imino proton exchange: detection of poly(rU) and of a dendrimer-poly(rU) model for nucleic acid delivery and pharmacology. AB - It is shown that the exchange properties of the imino and hydroxyl protons of polyuridilic acid (poly(rU)) allow use of this compound as a chemical-exchange saturation transfer (CEST) contrast agent. A proton/proton sensitivity enhancement factor of over 5000 per imino proton allowed the detection of a few micromolar of polymer (2000 uridine units; 644 kD) with a 50% change in the water signal. The enhancement factor would increase further at even lower concentrations, opening up the submicromolar range. When poly(rU) was complexed to a dendrimer carrying 250 positive charges, the stoichiometry was approximately one RNA for 10 dendrimers. The sensitivity enhancement was reduced but remained large (2300/imino proton), bringing enhanced CEST visibility to the dendrimers. The net charge of the complex was positive, suggesting that the complexed dendrimers would still interact with cell membranes, and that the RNA-dendrimer complex could provide a model for a gene delivery system with good CEST visibility. PMID- 12768578 TI - Method to determine in vivo the relaxation time T1 of hyperpolarized xenon in rat brain. AB - The magnetic polarization of the stable (129)Xe isotope may be enhanced dramatically by means of optical techniques and, in principle, hyperpolarized (129)Xe MRI should allow quantitative mapping of cerebral blood flow with better spatial resolution than scintigraphic techniques. A parameter necessary for this quantitation, and not previously known, is the longitudinal relaxation time (T(1) (tissue)) of (129)Xe in brain tissue in vivo: a method for determining this is reported. The time course of the MR signal in the brain during arterial injection of hyperpolarized (129)Xe in a lipid emulsion was analyzed using an extended two compartment model. The model uses experimentally determined values of the RF flip angle and the T(1) of (129)Xe in the lipid emulsion. Measurements on rats, in vivo, at 2.35 T gave T(1) (tissue) = 3.6 +/- 2.1 sec (+/-SD, n = 6). This method enables quantitative mapping of cerebral blood flow. PMID- 12768577 TI - Receptor-mediated endocytosis of iron-oxide particles provides efficient labeling of dendritic cells for in vivo MR imaging. AB - Dendritic cells (DCs) function as antigen presenting cells in vivo and play a fundamental role in numerous diseases. New methods are described for high efficiency intracellular labeling of DCs with superparamagnetic iron-oxide (SPIO) utilizing a receptor-mediated endocytosis (RME) mechanism. Bone marrow-derived DCs or a fetal skin-derived DC line were incubated with SPIO conjugated to anti CD11c monoclonal antibody (mAb) under conditions favoring RME. These cells exhibited approximately a 50-fold increase in uptake relative to DCs incubated with SPIO without the mAb. Flow cytometry studies assaying cell surface markers showed a down-modulation of CD11c, but no other changes in phenotype. Immunological function of the DCs was unmodified by the labeling, as determined by cytokine secretion assays. The RME mechanism was confirmed using electron microscopy, endocytosis inhibition assays, and incubation experiments with SPIO conjugated to mAbs against accessory molecules that are not expressed on DCs. Labeled DCs were injected into murine quadriceps and monitored in vivo for several days using MR microimaging at 11.7 T. DCs were observed to remain within the muscle for >24 hr. The use of RME is an efficient way to label immune cells for in vivo MRI and can be applied to a wide variety of cell types. PMID- 12768579 TI - Microvascular BOLD contribution at 4 and 7 T in the human brain: gradient-echo and spin-echo fMRI with suppression of blood effects. AB - The BOLD signal consists of an intravascular (IV) and an extravascular (EV) component from both small and large vessels. Their relative contributions are dependent on field strength, imaging technique, and echo time. The IV and EV contributions were investigated in the human visual cortex at 4 and 7 T using spin-echo and gradient-echo BOLD fMRI with and without suppression of blood effects. Spin-echo acquisition suppresses EV BOLD from large veins and reflects predominantly blood T(2) changes and EV BOLD signal from small blood vessels. At a short echo time (32 ms), diffusion gradient-based suppression of blood signals resulted in a 75% and 20% decrease in spin-echo BOLD changes at 4 T and 7 T, respectively. However, at echo times (55-65 ms) approximating tissue T(2) typically used for optimal BOLD contrast, these gradients had much smaller effects at both fields, consistent with the decreasing blood T(2) with increasing field strength. Gradient-echo BOLD percent changes, with relatively long echo times at both fields, were virtually unaffected by gradients that attenuated the blood contribution because the EV BOLD surrounding both large and small vessels dominated. These results suggest that spin-echo BOLD fMRI at 4 and 7 T, with TE approximating tissue T(2), significantly reduces nonspecific mapping signals from large vessels and significantly accentuates microvasculature contributions. PMID- 12768580 TI - Fast measurement of relaxation times by steady-state free precession of 129Xe in carrier agents for hyperpolarized noble gases. AB - Hyperpolarized gases ((129)Xe and (3)He) are being used increasingly in both MRI and NMR spectroscopy studies. However, it has been shown that carrier agents are required to preserve the long relaxation times of gases in biological fluids. Optimized gas transport can be achieved through controlled T(1) and T(2) measurements of (129)Xe gas at equilibrium, using the steady-state free precession method (SSFP). The accuracy of the method was proven with the use of CuSO(4)-doped water samples and xenon dissolved in chloroform. The following T(1) and T(2) values were measured for xenon dissolved in a 30% intralipid emulsion: T(1) = 29 +/- 3 s; T(2) = 1.0 +/- 0.1 s. The values obtained in the intralipid emulsion contrast significantly with those obtained in conventional gas NMR experiments, in which it is commonly assumed that T(1) = T(2). This highlights the importance of obtaining accurate relaxation time measurements for medical applications of hyperpolarized gases. PMID- 12768581 TI - Magnetic resonance imaging of gastric cancer in Tff1 knock-out mice. AB - In this work, the use of MRI to stage gastric neoplasia in a mouse model of spontaneous gastric cancer is demonstrated. The methodology involves 1) the use of deuterated water ((2)H(2)O) to distend the stomach, and to provide negative contrast between the stomach and the lumen without inducing susceptibility-based field shifts; 2) GlucaGen to minimize motion artifacts that arise from peristalsis; and 3) Gd-DTPA to enhance contrast between the dysplasia/tumor and the normal wall. Initial images are presented from a Tff1 -/- homozygous knock out model of gastric cancer and a heterozygous C57BL6/J control mouse. There are clear differences between the two types of animals in the MR appearance of the stomach. The distended stomach of the control mouse has a smooth perimeter and a thin wall, and an absence of nodules. The stomach of the Tff1 -/- mouse demonstrates a thickened wall; a jagged, irregular surface; and protruding nodules that are bright after injection of Gd-DTPA. PMID- 12768582 TI - In vivo assessment of macromolecular content in articular cartilage of the goat knee. AB - Loss of proteoglycans (PGs) from the extracellular matrix of cartilage is an early event of osteoarthritis. The capability of Gd(DTPA)(2-)-enhanced MRI to quantitatively assess PG content was explored in a goat model of cartilage degeneration. Partial to total PG depletion was induced by an intraarticular injection of papain 1 day prior to the MRI session. A close correlation was found between the extent of the PG loss and the Gd(DTPA)(2-)-induced T(1) decrease. Papain-induced PG depletion was confirmed by post-mortem histological and biochemical assessments. A 2-hr delay after Gd(DTPA)(2-) injection was found to be optimal for an accurate quantitation of the cartilage defect. A series of knee flexions were performed post-Gd(DTPA)(2-) injection to facilitate penetration of the contrast agent into cartilage. However, DeltaT(1)'s observed in cartilage of exercised goat knees were not affected by papain or IL1beta pretreatment. Therefore, as long as a preinjection T(1) map was obtained, the Gd(DTPA)(2-) enhanced MRI technique provided good sensitivity in detecting partial loss of PG in articular cartilage. This was true only when the animal was maintained in a resting state during diffusion of the Gd(DTPA)(2-). This approach is of particular interest for long-term evaluations of cartilage degeneration and regeneration. PMID- 12768583 TI - Macrophage infiltration into the rat knee detected by MRI in a model of antigen induced arthritis. AB - Three-dimensional (3D) MR images were obtained from the knees of rats in a model of antigen-induced arthritis, elicited by the intraarticular administration of methylated bovine serum albumin (mBSA) to previously immunized rats. Superparamagnetic particles of iron oxide (SPIO) were administered i.v. 24 hr before each imaging session. Starting 4 days postantigen injection, images from arthritic knees exhibited distinctive signal attenuation in the synovium. This signal attenuation was significantly smaller in knees from animals treated with dexamethasone, a glucocorticosteroid, and completely absent in contralateral knees that had been challenged with vehicle. A significant negative correlation was found between the MRI signal intensity in the synovium and the histologically determined iron content in macrophages located in the same region. These results suggest the feasibility of detecting macrophage infiltration into the knee synovium in this model of antigen-induced arthritis by labeling the cells with SPIO. This readout could provide an early marker of disease progression, before more aggressive changes like cartilage and bone erosion take place. Monitoring early changes associated with arthritis can have an impact in preclinical studies by shortening the duration of the experimental period and by facilitating the investigation of novel immunomodulatory therapies acting on macrophages. Also, the approach can be potentially adapted to clinical studies. PMID- 12768584 TI - Fast measurement of intracardiac pressure differences with 2D breath-hold phase contrast MRI. AB - Intracardiovascular blood pressure differences can be derived from velocity images acquired with phase-contrast (PC) MRI by evaluating the Navier-Stokes equations. Pressure differences within a slice of interest can be calculated using only the in-plane velocity components from that slice. This rapid exam is proposed as an alternative to the lengthy 3D velocity imaging exams. Despite their good spatial coverage, the 3D exams are prone to artifacts and errors from respiratory motion and insufficient temporal resolution, and are unattractive in the clinical setting due to their excessive scan times (>10 min of free breathing). The proposed single-slice approach requires only one or two breath holds of acquisition time, and the velocity data can be processed for the calculation of pressure differences online with immediate feedback. The impact of reducing the pressure difference calculation to two dimensions is quantified by comparison with 3D data sets for the case of blood flow within the cardiac chambers. The calculated pressure differences are validated using high-fidelity pressure transducers both in a pulsatile flow phantom and in vivo in a dog model. There was excellent agreement between the transducer and PC-MRI results in all of the studies. PMID- 12768585 TI - Measuring the arterial input function with gradient echo sequences. AB - The measurement of the arterial input function by use of gradient echo sequences was investigated by in vitro and in vivo experiments. First, calibration curves representing the influence of the concentration of Gd-DTPA on both the phase and the amplitude of the MR signal were measured in human blood by means of a slow infusion experiment. The results showed a linear increase in the phase velocity and a quadratic increase in DeltaR(*) (2) as a function of the Gd-DTPA concentration. Next, the resultant calibration curves were incorporated in a partial volume correction algorithm for the arterial input function determination. The algorithm was tested in a phantom experiment and was found to substantially improve the accuracy of the concentration measurement. Finally, the reproducibility of the arterial input function measurement was estimated in 16 patients by considering the input function of the left and the right sides as replicate measurements. This in vivo study showed that the reproducibility of the arterial input function determination using gradient echo sequences is improved by employing a partial volume correction algorithm based on the calibration curve for the contrast agent used. PMID- 12768586 TI - Circular spectrum mapping for intravoxel fiber structures based on high angular resolution apparent diffusion coefficients. AB - A method is presented for mapping intravoxel fiber structures using spectral decomposition onto a circular distribution of measured apparent diffusion coefficients (ADCs). The zeroth-, second-, and fourth-order harmonic components of the ADC distribution on the circle spanned by the major and median eigenvectors of the diffusion tensor can be used to provide quantitative indices for isotropic, linear, and fiber-crossing diffusion, respectively. A diffusion weighted MRI technique with 90 encoding orientations was implemented to estimate the circular ADC distribution and calculate the circular spectrum. A digital phantom was used to simulate various diffusion patterns. Comparisons were made between the circular spectrum and regular DTI-based index maps. The results indicated that the zeroth- and second-order circular spectrum maps exhibited a strong consistency with the DTI-based mean diffusivity and linear indices, respectively, and the fourth-order circular spectrum map was able to identify the fiber crossings. MRI experiments were performed on seven healthy human brains using a 3T scanner. The in vivo fourth-order maps showed significantly higher densities in several brain regions, including the corpus callosum, cingulum bundle, superior longitudinal fasciculus, corticospinal tract, and middle cerebellar peduncle, which indicated the existence of fiber crossings in these regions. PMID- 12768587 TI - Optimized spiral imaging for measurement of myocardial T2 relaxation. AB - Microcirculation oxygen levels and blood volumes should be reflected in measurements of myocardial T(2) relaxation. This work describes the optimization of a spiral imaging strategy for robust myocardial T(2) measurement to minimize the standard deviation of T(2) measurement (sigmaT(2)). Theoretical and experimental studies of blurring at muscle/blood interfaces enabled the derivation of parameter sets which reduce sigma T(2) to the level of 5%. T(2) relaxation mapping within healthy volunteers provided estimation of residual sigmaT(2) within the optimized technique. The standard deviation in T(2) measurement across regions of interest (ROIs) in different locations is about 9%. The standard deviation in T(2) measurement in an ROI across different time points is about 5%. PMID- 12768588 TI - Increased diffusion sensitivity with hyperechos. AB - It is shown that the introduction of a 180 degrees refocusing pulse into a standard diffusion weighted stimulated echo sequence is equivalent to the simplest hyperecho sequence with identical diffusion weighting but equal or greater signal-to-noise (SNR) and thus equal or greater diffusion contrast. For high b-value imaging, the hyperecho sequence thus possesses the high diffusion contrast in the presence of small T(1)/T(2) ratios characteristic of stimulated echo sequences but with less than the 50% loss in SNR that is associated with the stimulated echo. For low b-value imaging, the hyperecho signal converges to that of the standard spin echo. The advantages of the two-pulse diffusion weighted hyperecho sequence are demonstrated theoretically. Experimental results are shown in the application to high angular resolution diffusion encoding (HARD) in normal human brain. PMID- 12768589 TI - Extended field-of-view imaging with table translation and frequency sweeping. AB - A new method for MRI of an extended field of view (FOV) has been developed and validated. The method employs concurrent MR data acquisition and patient table motion. Table motion-induced image artifacts are minimized by sweeping the frequency of the receiver at a rate matching the table's speed. Multiple regional images are collected and combined to reconstruct the full FOV. The imaging parameters and table speed are chosen to ensure that each regional image of the subject is collected while the corresponding anatomy is in the useable imaging volume of the scanner. Additional strategies are applied to further reduce field inhomogeneity-induced artifacts, especially distortions due to gradient field nonlinearity. The method is robust and can be easily incorporated into most multislice 2D and volumetric 3D imaging pulse sequences. It is anticipated that this technique will be useful for a variety of applications, including angiographic runoffs, whole-body screening, and short-magnet imaging. PMID- 12768590 TI - Reduction of spectral ghost artifacts in high-resolution echo-planar spectroscopic imaging of water and fat resonances. AB - Echo-planar spectroscopic imaging (EPSI) can be used for fast spectroscopic imaging of water and fat resonances at high resolution to improve structural and functional imaging. Because of the use of oscillating gradients during the free induction decay (FID), spectra obtained with EPSI are often degraded by Nyquist ghost artifacts arising from the inconsistency between the odd and even echoes. The presence of the spectral ghost lines causes errors in the evaluation of the true spectral lines, and this degrades images derived from high-resolution EPSI data. A technique is described for reducing the spectral ghost artifacts in EPSI of water and fat resonances, using echo shift and zero-order phase corrections. These corrections are applied during the data postprocessing. This technique is demonstrated with EPSI data acquired from human brains and breasts at 1.5 Tesla and from a water phantom at 4.7 Tesla. Experimental results indicate that the present approach significantly reduces the intensities of spectral ghosts. This technique is most useful in conjunction with high-resolution EPSI of water and fat resonances, but is less applicable to EPSI of metabolites due to the complexity of the spectra. PMID- 12768591 TI - Error reduction and parameter optimization of the TAPIR method for fast T1 mapping. AB - A methodology is presented for the reduction of both systematic and random errors in T(1) determination using TAPIR, a Look-Locker-based fast T(1) mapping technique. The relations between various sequence parameters were carefully investigated in order to develop recipes for choosing optimal sequence parameters. Theoretical predictions for the optimal flip angle were verified experimentally. Inversion pulse imperfections were identified as the main source of systematic errors in T(1) determination with TAPIR. An effective remedy is demonstrated which includes extension of the measurement protocol to include a special sequence for mapping the inversion efficiency itself. PMID- 12768592 TI - Selective arterial spin labeling (SASL): perfusion territory mapping of selected feeding arteries tagged using two-dimensional radiofrequency pulses. AB - To date, most perfusion magnetic resonance imaging (MRI) methods using arterial spin labeling (ASL) have employed slab-selective inversion pulses or continuous labeling within a plane in order to obtain maps derived from all major blood vessels entering the brain. However, there is great potential for gaining additional information on the territories perfused by the major vessels if individual feeding arteries could be tagged. This study demonstrates noninvasive arterial perfusion territory maps obtained using two-dimensional (2D) selective inversion pulses. This method is designated "selective ASL" (SASL). The SASL method was used to tag the major arteries below the circle of Willis. A combination of 2D selective tagging and multislice readout allows perfusion territories to be clearly visualized, with likely applications to cerebrovascular disease and stroke. PMID- 12768593 TI - Anisotropic noise propagation in diffusion tensor MRI sampling schemes. AB - The subject of this study is the controversial choice of directions in diffusion tensor MRI (DT-MRI); specifically, the numerical algebra related to this choice. In DT-MRI, apparent diffusivities are sampled in six or more directions and a least-squares equation is solved to reconstruct the diffusion tensor. Numerical characteristics of the system are considered, in particular the condition number and normal matrix, and are shown to be dependent on the relative orientation of the tensor with respect to the laboratory frame. As a consequence, noise propagation can be anisotropic. However, the class of icosahedral direction schemes is an exception, and icosahedral directions have the same condition number and normal matrix for direction encoding as the ideal scheme with an infinite number of directions. This normal matrix and its condition number are rotationally invariant. Numerical simulations show that for icosahedral schemes with 30 directions the standard deviation of the fractional anisotropy is both low and nearly independent of fiber orientation. The recommended choice of directions for a DT-MRI experiment is therefore the icosahedral set of directions with the highest number of directions achievable in the available time. PMID- 12768594 TI - Novel ROC-type method for testing the efficiency of multivariate statistical methods in fMRI. AB - The receiver operating characteristic (ROC) method is a useful and popular tool for testing the efficiency of various diagnostic tests applicable to functional MRI (fMRI) data. Typically, the diagnostic tests are applied on simulated and pseudo-human fMRI data, and the area under the ROC curve is used as a measure of the efficiency of the diagnostic test. The effectiveness of such a method depends on how well the simulated data approximate the real data. For multivariate statistical methods, however, this technique is usually inadequate, as the spatial dependence among voxels is ignored for simulated data. In this work a modified ROC method using real fMRI data with a broader scope is proposed. This method can be applied to most fMRI postprocessing techniques, including multivariate analyses such as canonical correlation analysis (CCA). Also, the relationship of the modified ROC method with the conventional ROC method is discussed in detail. PMID- 12768595 TI - Image-based tracking of optically detunable parallel resonant circuits. AB - In this work strategies for the robust localization of parallel resonant circuits are investigated. These strategies are based on the subtraction of two images, which ideally differ in signal intensity at the positions of the devices only. To modulate their signal amplification, and thereby generate the local variations, the parallel resonant circuits are alternately detuned and retuned during the acquisition. The integration of photodiodes into the devices permits their fast optical switching. Radial and spiral imaging sequences are modified to provide the data for the two images in addition to those for a conventional image in the same acquisition time. The strategies were evaluated by phantom experiments with stationary and moving catheter-borne devices. In particular, rapid detuning and retuning during the sampling of single profiles is shown to lead to a robust localization. Moreover, this strategy eliminates most of the drawbacks usually associated with image-based tracking, such as low temporal resolution. Image based tracking may thus become a competitive (if not superior) alternative to projection-based tracking of parallel resonant circuits. PMID- 12768596 TI - Spectral fitting: the extraction of crucial information from a spectrum and a spectral image. AB - A highly accurate line-width simulation computer program is used that can account for both high amplitude and frequency of the Zeeman modulation in an electron paramagnetic resonance (EPR) experiment. This allows for the overmodulation of EPR lines to increase signal-to-noise ratio (SNR) in EPR spectra and spectroscopic images, without any sacrifice in the determination of the intrinsic line width (1/gamma. T(2e)). The technique was applied to continuous-wave EPR spectroscopic images of a narrow, single-line trityl spin probe wherein a full EPR spectrum was extracted from each 3D spatial voxel. Typical improvements are a three- to fivefold increase in SNR in the high-gradient projections in the image and a reduction in the standard deviation (SD), by a factor of 3, of the line widths in the low-gradient domain. This method is a general one that is also applicable to the analysis of conventional (14)N or (15)N nitroxide spin probes. PMID- 12768597 TI - Mapping ischemic risk region and necrosis in the isolated heart using EPR imaging. AB - Reperfusion of ischemic tissue is a common event in the treatment of heart attack and stroke. To study disease pathogenesis, methods are required to measure tissue perfusion and area at risk, as well as localized regions of injury. While histology can provide this information, its destructive nature precludes assessment of time course. Thus, there is a critical need for a noninvasive technique to obtain this information. To map myocardial redox state as a possible index of cellular ischemia and viability, electron paramagnetic resonance (EPR) imaging experiments were performed on isolated rat hearts before and after the onset of regional ischemia using nitroxide spin labels. With coronary artery occlusion, the EPR images clearly showed the risk region as a void of lower intensity that reversed upon reperfusion. The extent of risk region in the heart was similar in EPR imaging and histological measurements. The unique information obtained regarding the time course of changes in redox metabolism of the risk region and normal myocardium can provide important insights regarding the mechanisms of myocardial injury during and following ischemia. PMID- 12768599 TI - Improved preparation of chick embryonic samples for magnetic resonance microscopy. AB - Previous work demonstrated the power of three-dimensional (3D) magnetic resonance microscopy (MRM) to follow complicated morphologic development in the embryonic cardiovascular system. In this study we describe a new dual-contrast method for specimen preparation that combines perfusion fixation and immersion in fixative with macro- and small molecular gadolinium agents to provide enhanced definition of both the heart wall and chamber. MRM was performed at 9.4 T with image resolutions of 25, 31, and 50 microm isotropic voxels for three stages of chick embryos (day 4, day 5.5, and day 9), and compared to histological sections of the same embryos. The results show considerable improvement of image quality over previous efforts, with better signal-to-noise ratio (SNR) and contrast between the cardiac chamber and myocardial wall. Excellent correlation was shown between the MRM images and histological sections. Thus, 3D high-resolution MRM in combination with the dual-contrast technique is useful for acquiring quantitative 3D morphologic data regarding heart development. PMID- 12768598 TI - MRI-guided focused ultrasound surgery in the brain: tests in a primate model. AB - MRI-guided focused ultrasound was tested in the brains of rhesus monkeys. Locations up to 4.8 cm deep were targeted. Focal heating was observed in all cases with MRI-derived temperature imaging. Subthreshold heating was observed at the focus when the ultrasound beam was targeted with low power sonications, and in the ultrasound beam path during high-power exposures. Lethal temperature values and histologically confirmed tissue damage were confined to the focal zone (e.g., not in the ultrasound beam path), except when the focus was close to the bone. In that case, damage to the neighboring brain tissue was observed. Focal lesions were observed on histological examination and, in some cases, in MR images acquired immediately after the ultrasound exposures. The capabilities demonstrated in this study will be of benefit for clinical ultrasound therapies in the brain. PMID- 12768600 TI - B1AC-MAMBA: B1 array combined with multiple-acquisition micro B0 array parallel magnetic resonance imaging. AB - The combination of an in-plane B(1) sensitivity encoding (SENSE) technique with a simultaneous multiple-slice B(0) field step technique (multiple-acquisition micro B(0) array (MAMBA)) has produced high scan time reduction factors (R < or = 8). In this study, two slices were acquired simultaneously in combination with x2 and x4 SENSE in-plane encoding using a MAMBA stepped B(0) field coil inside a four channel phased-array coil system. Experiments were performed on a 1.5 T Infinion system (Philips Medical Systems, Cleveland, OH). The signal-to-noise ratio (SNR) was reduced with higher R factors, as was expected from the reduced number of acquisitions used to create the unaliased images. The combination of SENSE and MAMBA offers great promise for reducing scan times through parallel acquisition while at the same time reducing the number of RF channels required by a factor equal to the number of field steps employed. The B(1) array combined with MAMBA (B(1)AC-MAMBA) technique is applicable when the length of an object is much greater than its diameter, as in scanning limbs or in whole-body screening for disease. PMID- 12768602 TI - Protein-folding kinetics and mechanisms studied by pulse-labeling and mass spectrometry. AB - The "protein-folding problem" refers to the question of how and why a denatured polypeptide chain can spontaneously fold into a compact and highly ordered conformation. The classical description of this process in terms of reaction pathways has been complemented by models that describe folding as a biased conformational diffusion on a multidimensional energy landscape. The identification and characterization of short-lived intermediates provide important insights into the mechanism of folding. Pulsed hydrogen/deuterium exchange (HDX) methods are among the most powerful tools for studying the properties of kinetic intermediates. Analysis of pulse-labeled proteins by mass spectrometry (MS) provides information that is complementary to that obtained in nuclear magnetic resonance (NMR) studies; NMR data represent an average of entire protein ensembles, whereas MS can detect co-existing protein species. MS-based pulse-labeling experiments can distinguish between folding scenarios that involve parallel pathways, and those where folding is channeled through obligatory intermediates. The proteolytic digestion/MS technique provides spatially resolved information on the HDX pattern of folding intermediates. This method is especially important for proteins that are too large to be studied by NMR. Although traditional pulsed HDX protocols are based on quench-flow techniques, it is also possible to use electrospray (ESI) MS to analyze the reaction mixture on line and "quasi-instantaneously" after labeling. This approach allows short-lived protein conformations to be studied by their HDX level, their ESI charge-state distribution, and their ligand-binding state. Covalent labeling of free cysteinyl residues provides an alternative approach to pulsed HDX experiments. Another promising development is the use of synchrotron X-rays to induce oxidation at specific sites within a protein for studying their solvent accessibility during folding. PMID- 12768603 TI - Subcellular proteomics. AB - The step from the analysis of the genome to the analysis of the proteome is not just a matter of numerical complexity in terms of variants of gene products that can arise from a single gene. A significant further level of complexity is introduced by the supramolecular organization of gene products because of protein protein interactions or targeting of proteins to specific subcellular structures. There is currently no single proteome analysis strategy that can sufficiently address all levels of the organization of the proteome. To approach an appropriate analytical complement for the interrogation of the proteome at all of the levels at which it is organized, there emerges the need for a whole arsenal of proteomics strategies. The proteome analysis at the level of subcellular structures (that can be enriched by subcellular fractionation) represents an analytical strategy that combines classic biochemical fractionation methods and tools for the comprehensive identification of proteins. Among the key potentials of this strategy is the capability to screen not only for previously unknown gene products but also to assign them, along with other known, but poorly characterized gene products, to particular subcellular structures. Furthermore, the analysis at the subcellular level is a prerequisite for the detection of important regulatory events such as protein translocation in comparative studies. This review is meant to give an overview on recent key studies in the field of proteome analysis at the level of subcellular structures, and to highlight potentials and requirements. PMID- 12768604 TI - Reactions of polypeptide ions with electrons in the gas phase. AB - Reactions of electrons in the energy range below 70 eV with polypeptide cations and anions are reviewed, as well as their applications for the structural analysis of polypeptides. At very low energies (3)-beta-D-glucan, and a questionnaire survey were carried out. Blood data was restricted to MRFs 3, 6, and 9 (45 workers). Blood sampling investigated differential cell counts, erythrocyte sedimentation rate (ESR), and immunoglobulin (Ig)E. RESULTS: Workers exposed to higher amounts of endotoxin and (1-->3)-beta-D-glucan had an increased risk for respiratory symptoms as compared to those with lower exposure. Stomach problems was associated to higher (1-->3)-beta-D-glucan exposure. MRF 3 had a higher (1-->3)-beta-d-glucan exposure compared to 6 and 9, and respiratory symptoms, unusual tiredness, and vomiting were reported more often in MRF 3. Monocyte numbers and ESR were significantly decreased in MRF 3 compared to MRF 6 and 9, but all measured values were within normal ranges. CONCLUSIONS: The results suggest that MRF workers exposed to higher levels of endotoxin and (1- >3)-beta-D-glucan at their work sites exhibit various work-related symptoms, and that the longer a worker is in the MRF environment, the more likely he is to become affected by various respiratory and gastrointestinal symptoms. PMID- 12768609 TI - Beryllium-stimulated neopterin as a diagnostic adjunct in chronic beryllium disease. AB - BACKGROUND: The diagnosis of chronic beryllium disease (CBD) relies on the beryllium lymphocyte proliferation test (BeLPT) to demonstrate a Be specific immune response. This test has improved early diagnosis, but cannot discriminate beryllium sensitization (BeS) from CBD. We previously found high neopterin levels in CBD patients' serum and questioned whether Be-stimulated neopterin production by peripheral blood cells in vitro might be useful in the diagnosis of CBD. METHODS: CBD, BeS, Be exposed workers without disease (Be-exp) normal controls and sarcoidosis subjects were enrolled. Peripheral blood mononuclear cells (PBMN) were cultured in the presence and absence of beryllium sulfate. Neopterin levels were determined from cell supernatants by enzyme linked immunosorbent assay (ELISA). Clinical evaluation of CBD subjects included chest radiography, pulmonary function testing, exercise testing, and the BeLPT. RESULTS: CBD patients produced higher levels of neopterin in both unstimulated and Be stimulated conditions compared to all other subjects (P < 0.0001). Unstimulated neopterin mononuclear cell levels overlapped among groups, however, Be-stimulated neopterin levels in CBD showed little overlap. Using a neopterin concentration of 2.5 ng/ml as a cutoff, Be-stimulated neopterin had a sensitivity of 80% and specificity of 100% for CBD and was able to differentiate CBD from BeS. Be stimulated neopterin was inversely related to measures of pulmonary function, exercise capacity, and gas exchange. CONCLUSIONS: Neopterin may be a useful diagnostic adjunct in the non-invasive assessment of CBD, differentiating CBD from BeS. Further studies will be required to determine how it performs in workplace screening. PMID- 12768610 TI - Monitoring of DNA damage in foundry and pottery workers exposed to silica by the alkaline comet assay. AB - BACKGROUND: Workers in foundry and pottery are exposed to a mixture of chemicals and silica, which is suspected to cause genetic alterations. METHODS: To investigate the potential hazard associated with the occupational exposure to silica, DNA damage in the peripheral lymphocytes of 30 foundry and 22 pottery workers were examined by the alkaline single-cell gel electrophoresis or Comet Assay, and compared to 52 healthy subjects with no history of occupational silica or chemical exposure. RESULTS: The DNA damage observed in the lymphocytes of both foundry and pottery workers was significantly higher than that in their controls. Cigarette smoking was also related to DNA damage since the DNA damage observed in smoking silica-exposed workers compared with the non-smoking workers was significantly higher. CONCLUSIONS: Occupational exposure of silica from foundry and pottery workplaces has been associated with the increased DNA damage and smoking which represents an additional risk factor and must be avoided. PMID- 12768611 TI - Occupational asthma in the city of Sao Paulo, 1995-2000, with special reference to gender analysis. AB - BACKGROUND: Occupational asthma (OA) is the leading occupational respiratory disease in the area of Sao Paulo Municipality. Understanding its characteristics can provide useful information for better interventions. METHODS: From a joint registry of OA from five outpatient public clinics for occupational respiratory diseases, data on occupation, with job titles corrected by an expert evaluation, agents, exposure and symptom duration, and spirometry were analyzed by gender. Data on employment for Sao Paulo Municipality were used to calculate expected ratios for men/women in broad economical sectors. RESULTS: Three hundred and ninety four OA cases were reported with a ratio of men/women of 1.5. Women had significantly shorter exposure duration (5.6 +/- 5.2 vs. 8.9 +/- 9.0 years, P = 0.0005), shorter symptoms duration (2.6 +/- 3.7 vs. 3.2 +/- 3.5 years, P = 0.1270), and higher prevalence of previous atopy (27.0 vs. 18.4%, P = 0.0485). The main occupations related to OA cases were cleaning activities, working in plastics manufacture and in chemical and pharmaceutical plants. Women reported significantly more exposures to cleaning products, biologic agents, and textile fibers, whereas men reported more exposures to isocyanates, metal dusts and fumes, oil mists, wood dusts and anhydrides. Women presented an increased risk of OA in the service sector (odds ratio (OR) = 1.77, confidence interval (CI) = 1.61 1.96). CONCLUSIONS: Cleaning services was the main reported occupation and cleaning products the main reported agents. Women had significantly shorter exposure duration possibly due to difference in exposures, previous atopy, and behavior. There was an excess of cases of women in the service sector. PMID- 12768613 TI - Environmental risk factors and work-related lower respiratory symptoms in 80 office buildings: an exploratory analysis of NIOSH data. AB - BACKGROUND: We evaluated relationships between lower respiratory symptoms and risk factors for microbiological contamination in office buildings. METHODS: The National Institute for Occupational Safety and Health collected data from 80 office buildings during standardized indoor environmental health hazard evaluations. Present analyses included lower respiratory symptom-based outcome definitions and risk factors for potential microbiologic contamination. Multivariate logistic regression models for selected outcomes identified key risk factors. RESULTS: Adjusted odds ratios (95% confidence intervals) for "at least three of four work-related lower respiratory symptoms" were, for debris in ventilation air intake, 2.0 (1.0-3.9), and for poor drainage in air-conditioning drip pans, 2.6 (1.3-5.2). Adjusted associations with risk factors were consistently stronger for outcomes requiring both multiple symptoms and improvement away from work, and somewhat stronger among diagnosed asthmatics. CONCLUSIONS: Moisture and debris in ventilation systems, possibly by supporting microbiologic growth, may increase adverse respiratory effects, particularly among asthmatics. Data from more representative buildings are needed to confirm these findings. PMID- 12768612 TI - Be the fairest of them all: challenges and recommendations for the treatment of gender in occupational health research. AB - BACKGROUND: Both women's and men's occupational health problems merit scientific attention. Researchers need to consider the effect of gender on how occupational health issues are experienced, expressed, defined, and addressed. More serious consideration of gender-related factors will help identify risk factors for both women and men. METHODS: The authors, who come from a number of disciplines (ergonomics, epidemiology, public health, social medicine, community psychology, economics, sociology) pooled their critiques in order to arrive at the most common and significant problems faced by occupational health researchers who wish to consider gender appropriately. RESULTS: This paper describes some ways that gender can be and has been handled in studies of occupational health, as well as some of the consequences. The paper also suggests specific research practices that avoid errors. Obstacles to gender-sensitive practices are considered. CONCLUSIONS: Although gender-sensitive practices may be difficult to operationalize in some cases, they enrich the scientific quality of research and should lead to better data and ultimately to well-targeted prevention programs. PMID- 12768614 TI - Testicular cancer among Swedish pulp and paper workers. AB - BACKGROUND: The incidence of testicular cancer has increased in recent decades. The aims of the present study were to elucidate whether Swedish paper and pulp mill workers had an increased incidence of testicular cancer, and to investigate whether certain occupational groups within the pulp and paper mill workforce were at increased risk. METHODS: The study was based on the Swedish Cancer Environment Register, which links the incidence of cancer for the period 1971-1990 with 1960 and 1970 National Census data on specific industries and occupations for all employed subjects in Sweden. RESULTS: Among maintenance workers employed both in 1960 and in 1970 in paper mills there was an increased risk for testicular cancer (standardized incidence ratio (SIR) 7.4, 95% confidence intervals (CI) 1.5-22), especially for seminomas (SIR 10.1, 95% CI 2.1-29). Maintenance workers were also at increased risk when analyzing workers employed in 1960, 1970, or both years. CONCLUSIONS: This study indicates an increased risk for testicular cancer, especially seminomas, among maintenance workers, but not among process workers, in Swedish paper mills. PMID- 12768615 TI - Occupational dermatoses among Polish private farmers, 1991-1999. AB - BACKGROUND: Little is known about work-related skin diseases among Polish self employed farmers. In the National Register of Occupational Diseases, private farmers are placed in one category together with other agricultural workers, despite fundamental differences in compensation legislation and separate insurance institutions. The Agricultural Social Insurance Fund reports only on numbers of compensated cases. The aim of this study was to create reliable statistics on occupational dermatoses among private farmers. METHODS: All cases of work-related skin diseases diagnosed from 1991 to 1999 were included in the statistics. Compensation records of the Agricultural Social Insurance Fund were analyzed for diagnoses, causative factors, and health impairment of the skin. RESULTS: The first occupational dermatosis was registered in 1992. Until the end of 1999, there were 101 cases (63 women and 38 men). The incidence rose from 0.006/10000/year in 1992 to 0.189/10000/year in 1999. Allergic contact dermatitis was the most common diagnosis (86%), followed by infectious skin diseases (10%), irritant contact dermatitis (3%), and urticaria (2%). The most frequently identified causative factors were plant dusts (38%), animal allergens (36%), metals (29%), pesticides (18%), and rubber chemicals (15%). The median impairment due to skin disease was 20% (range 2-36%). CONCLUSIONS: Since the introduction of workers' compensation, the numbers of occupational dermatoses diagnosed in Polish private farmers have increased rapidly. However, compared to countries with a longer experience in this field, these figures remain low, probably due to low detection of these diseases. PMID- 12768616 TI - Cohort mortality study of employees at the Celanese plant in Pampa, Texas. AB - BACKGROUND: The mortality experience of chemical workers from the Pampa, Texas Celanese Ltd. plant through 1991 has been previously reported. This study updates that effort and presents an additional seven years of data and follow-up that provided 268 additional cohort members and 71 new deaths. METHODS: Mortality was determined through December 31, 1998, the period for which the National Death Index could provide information on deaths. The mortality experience of Celanese Ltd. employees was compared to that of the general population of the United States. All cause and cause-specific standardized mortality ratios (SMRs) were calculated. RESULTS: For the entire cohort, several causes of death including all causes (SMR = 65.9; 95% confidence interval (CI) = 56.9-76.0), heart disease (SMR = 69.1; 95% CI = 53.6-87.8), and all malignant neoplasms (SMR = 74.2; 95% CI = 56.3-95.9) were significantly less than expected. SMRs were also calculated separately for white men, non-white men, and women. Unlike the initial study, the SMR for prostate cancer among white males was not significantly elevated (SMR=176.1; 95% CI = 76.0-347.0) and was much lower than the prior SMR of 330.4. This suggests that the initial findings, which were based on a small number of deaths, were likely due to chance. CONCLUSIONS: Overall, Celanese Ltd. employees from the Pampa plant have significantly lower than expected SMRs for several causes of death, and no causes of death that are significantly elevated. PMID- 12768617 TI - Indirect lead exposure among children of radiator repair workers. AB - BACKGROUND: Secondary exposure to lead has been identified as a public health problem since the late 1940s; we investigate the risk of lead exposure among families of radiator repair workers. METHODS: A sample of the wives and children, aged 6 months to 6 years (exposed children) (n = 19), of radiator repair workers and a sample of children whose parents were not occupationally exposed to lead (non-exposed children) (n = 29) were matched for age and residence; their geometric mean blood lead levels are compared. Blood samples were obtained by the finger stick method and environmental dust samples by the wipe method; both were analyzed using a portable anodic stripping voltameter. RESULTS: Dust lead levels were significantly higher in the houses of exposed children (143.8 vs. 3.9 microg/g; P < 0.01). In crude analyses, the highest lead levels were observed among children whose fathers worked in home-based workshops (22.4 microg/dl)(n = 6). Children whose fathers worked in an external workshop (n = 13) also had high levels (14.2 microg/dl) (P < 0.01), while blood lead levels in non-exposed children were significantly lower (5.6 microg/dl)(P < 0.01). The observed differences remained significant after adjustment for age and gender. CONCLUSIONS: This study confirms that children of radiator repair workers are at increased risk of lead exposure and public health interventions are needed to protect them. PMID- 12768618 TI - Protein loading, elution, and resolution behavior in a novel device that integrates ultrafiltration and chromatographic separation. AB - Hollow fiber membranes and chromatographic resin beads are commonly employed in a variety of bioseparation processes. A new class of integrated separation devices is being studied in which the shell side of a hollow fiber device is filled with adsorbents/chromatographic resin beads. Such devices and the corresponding separation methods integrate feed broth clarification by the microfiltration/ultrafiltration membrane with bioproduct purification by the shell-side resin beads either as an adsorbent or as beads in elution chromatography. A mathematical model has been developed for the prediction of the chromatographic behavior of such an integrated device. Simulations have been done to study the effects of axial dispersion, feed flow rate, water permeation rate, fiber packing density, and void fraction. Numerical solutions were obtained by solving the governing equations. This model can reasonably describe the concentration profiles as well as the breakthrough and elution behaviors in the integrated device. PMID- 12768619 TI - Identification and comparison of aerobic and denitrifying polyphosphate accumulating organisms. AB - Two laboratory-scale sequencing batch reactors (SBRs) were operated for enhanced biological phosphorus removal (EBPR) in alternating anaerobic-aerobic or alternating anaerobic-anoxic modes, respectively. Polyphosphate-accumulating organisms (PAOs) were enriched in the anaerobic-aerobic SBR and denitrifying PAOs (DPAOs) were enriched in the anaerobic-aerobic SBR. Fluorescence in situ hybridization (FISH) demonstrated that the well-known PAO, "Candidatus Accumulibacter phosphatis" was abundant in both SBRs, and post-FISH chemical staining with 4,6-diamidino-2-phenylindol (DAPI) confirmed that they accumulated polyphosphate. When the anaerobic-anoxic SBR enriched for DPAOs was converted to anaerobic-aerobic operation, aerobic uptake of phosphorus by the resident microbial community occurred immediately. However, when the anaerobic-aerobic SBR enriched for PAOs was exposed to one cycle with anoxic rather than aerobic conditions, a 5-h lag period elapsed before phosphorus uptake proceeded. This anoxic phosphorus-uptake lag phase was not observed in the subsequent anaerobic aerobic cycle. These results demonstrate that the PAOs that dominated the anaerobic-aerobic SBR biomass were the same organisms as the DPAOs enriched under anaerobic-anoxic conditions. PMID- 12768620 TI - Biomass/adsorbent electrostatic interactions in expanded bed adsorption: a zeta potential study. AB - Expanded bed adsorption is an integrated technology that allows the introduction of particle-containing feedstock without the risk of blocking the bed. The biomass particles contained in the feedstock have to be treated as an integral part of the process and potential interactions between suspended biomass and the adsorbent must be excluded during process design. Because the electrostatic forces dominate the interactions between the biomass and adsorbent, the zeta potential has been studied as a tool to characterize biomass/adsorbent electrostatic interactions. The zeta potentials of four types of biomass (yeast intact cells, yeast homogenate, Escherichia coli intact cells, and E. coli homogenate) and two types of ion exchanger were measured systematically at varying process conditions. Using the cell transmission index from biomass pulse response experiments as a parameter, the relations between zeta potential and the biomass/adsorbent interaction were evaluated. Combining the influences from zeta potential of adsorbent (zeta(a)), zeta potential of biomass (zeta(b)), and biomass size (d), parameter (-zeta(a)zeta(b)d) was found to be an appropriate indicator of the biomass/adsorbent interactions in expanded beds under various liquid-phase conditions for different types of biomass. The threshold value of parameter (-zeta(a)zeta(b)d) can be defined as 120 mV(2) microm for cell transmission of >90%, which means that systems with (-zeta(a)zeta(b)d) < 120 may have a considerable probability of forming stable expanded beds in a biomass suspension under the particular experimental conditions. PMID- 12768621 TI - Cadmium tolerance and antioxidative defenses in hairy roots of the cadmium hyperaccumulator, Thlaspi caerulescens. AB - Plant species capable of hyperaccumulating heavy metals are of considerable interest for phytoremediation and phytomining. This work aims to identify the role of antioxidative metabolism in heavy metal tolerance in the Cd hyperaccumulator, Thlaspi caerulescens. Hairy roots of T. caerulescens and the non-hyperaccumulator, Nicotiana tabacum (tobacco), were used to test the effects of high Cd environments. In the absence of Cd, endogenous activities of catalase were two to three orders of magnitude higher in T. caerulescens than in N. tabacum. T. caerulescens roots also contained significantly higher endogenous superoxide dismutase activity and glutathione concentrations. Exposure to 20 ppm (178 microM) Cd prevented growth of N. tabacum roots and increased hydrogen peroxide (H(2)O(2)) levels by a factor of five relative to cultures without Cd. In contrast, growth was maintained in T. caerulescens, and H(2)O(2) concentrations were controlled to low, nontoxic levels in association with a strong catalase induction response. Treatment of roots with the glutathione synthesis inhibitor, buthionine sulfoximine (BSO), exacerbated H(2)O(2) accumulation in Cd-treated N. tabacum, but had a relatively minor effect on H(2)O(2) levels and did not reduce Cd tolerance in T. caerulescens. Lipid peroxidation was increased by Cd treatment in both the hyperaccumulator and non hyperaccumulator roots. This work demonstrates that metal-induced oxidative stress occurs in hyperaccumulator tissues even though growth is unaffected by the presence of heavy metals. It also suggests that superior antioxidative defenses, particularly catalase activity, may play an important role in the hyperaccumulator phenotype of T. caerulescens. PMID- 12768622 TI - Mechanism of bactericidal and fungicidal activities of textiles covalently modified with alkylated polyethylenimine. AB - Our previous studies have led to a novel "nonrelease" approach to making materials bactericidal by covalently attaching certain moderately hydrophobic polycations to their surfaces. In the present work, this strategy is extended beyond the heretofore-used nonporous materials to include common woven textiles (cotton, wool, nylon, and polyester). Pieces of such cloths derivatized with N hexylated+methylated high-molecular-weight polyethylenimine (PEI) are strongly bactericidal against several airborne Gram-positive and Gram-negative bacteria. In contrast, the immobilized and N-alkylated PEIs of low molecular weight have only a weak, if any, bactericidal activity. These findings support a mechanism of the antibacterial action whereby high-molecular-weight and hydrophobic polycationic chains penetrate bacterial cell membranes/walls and fatally damage them. The bactericidal textiles prepared herein are lethal not only to pathogenic bacteria but to fungi as well. PMID- 12768623 TI - Method for generation of homogeneous multicellular tumor spheroids applicable to a wide variety of cell types. AB - Multicellular tumor spheroids (MCTS) are used as organotypic models of normal and solid tumor tissue. Traditional techniques for generating MCTS, such as growth on nonadherent surfaces, in suspension, or on scaffolds, have a number of drawbacks, including the need for manual selection to achieve a homogeneous population and the use of nonphysiological matrix compounds. In this study we describe a mild method for the generation of MCTS, in which individual spheroids form in hanging drops suspended from a microtiter plate. The method has been successfully applied to a broad range of cell lines and shows nearly 100% efficiency (i.e., one spheroid per drop). Using the hepatoma cell line, HepG2, the hanging drop method generated well-rounded MCTS with a narrow size distribution (coefficient of variation [CV] 10% to 15%, compared with 40% to 60% for growth on nonadherent surfaces). Structural analysis of HepG2 and a mammary gland adenocarcinoma cell line, MCF-7, composed spheroids, revealed highly organized, three-dimensional, tissue-like structures with an extensive extracellular matrix. The hanging drop method represents an attractive alternative for MCTS production, because it is mild, can be applied to a wide variety of cell lines, and can produce spheroids of a homogeneous size without the need for sieving or manual selection. The method has applications for basic studies of physiology and metabolism, tumor biology, toxicology, cellular organization, and the development of bioartificial tissue. PMID- 12768624 TI - Constitutive versus thermoinducible expression of heterologous proteins in Escherichia coli based on strong PR,PL promoters from phage lambda. AB - Constitutive and thermoinducible expression plasmids based on strong P(R),P(L) promoters from phage lambda were compared for production of TNF-alpha and its analogs under various conditions. Much higher accumulation of TNF was obtained in a constitutive system, so the wider applicability of such systems was studied. In constitutive systems, proteolytically susceptible proteins can be produced easily at low cultivation temperatures and the addition of expensive or toxic chemical inducers is not required. On the other hand, toxic proteins cannot be produced and selection pressure must be strictly maintained to ensure segregational stability of plasmids. Accumulation of TNF-alpha and various analogs at levels up to 25% of total soluble protein was repeatedly achieved, which was 2-3-fold higher than in a thermoinducible system. The stable behavior of the constitutive system in laboratory fermentors was also confirmed. We propose the constitutive system described here as a general model for many currently used expression systems containing strong but not completely repressed promoters. Such systems may be considered as constitutive ones with reduced promoter strengths, but still exhibiting all the intrinsic properties of constitutive expression systems. Although all modern expression systems are inducible, wider use of a constitutive system is evidently possible. PMID- 12768625 TI - Fermentation of corn stover to carboxylic acids. AB - This article describes countercurrent fermentation to anaerobically convert corn stover and pig manure to mixed carboxylic acids using a mixed culture of mesophilic microorganisms. Corn stover was pretreated with lime to increase digestibility. The Continuum Particle Distribution Model (CPDM) was used to simulate continuous fermentors based on data collected from batch experiments. This model saves considerable time in determining optimum operating conditions. For 80% corn stover/20% pig manure, the highest total carboxylic acid productivity was 1.81 g/(L of liquid. d) at a concentration of 21.4 g total acid/L. The highest total acid selectivity, yield, and conversion were 0.714 g total acid/g volatile solids (VS) digested, 0.550 g total acid/g VS fed, and 0.770 g VS digested/g VS fed, respectively, at a concentration of 16.0 g total acid/L. CPDM predicted the acid concentration and conversion within 13.4 and 11.6%, respectively. PMID- 12768626 TI - Separation of alpha-lactalbumin and beta-lactoglobulin using membrane ultrafiltration. AB - There is considerable commercial interest in the preparation of individual whey proteins as high-value food additives, nutraceuticals, and therapeutics. This study examined the use of membrane filtration for the separation of alpha lactalbumin and beta-lactoglobulin. Stirred cell filtration experiments were performed using both cellulosic and polyethersulfone membranes to determine the optimal pH, ionic strength, and filtration conditions. Selectivities of greater than 55 could be achieved at pH 5.5 and 50 mM ionic strength using a 30-kD cellulose membrane. A diafiltration process was then designed for the protein separation. A 16-diavolume filtration yielded beta-lactoglobulin as the retentate product with a purification factor of 100 and recovery of 90%. The alpha lactalbumin was recovered in the filtrate with a purification factor of more than 10 and nearly 99% yield. Model calculations were in good agreement with the experimental data. PMID- 12768627 TI - Modulation of translation-initiation in CHO-K1 cells by rapamycin-induced heterodimerization of engineered eIF4G fusion proteins. AB - Translation-initiation is a predominant checkpoint in mammalian cells which controls protein synthesis and fine-tunes the flow of information from gene to protein. In eukaryotes, translation-initiation is typically initiated at a 7 methyl-guanylic acid cap posttranscriptionally linked to the 5' end of mRNAs. Alternative cap-independent translation-initiation involves 5' untranslated regions (UTR) known as internal ribosome entry sites, which adopt a particular secondary structure. Translation-initiating ribosome assembly at cap or IRES elements is mediated by a multiprotein complex of which the initiation factor 4F (eIF4F) consisting of eIF4A (helicase), eIF4E (cap-binding protein), and eIF4G is a major constituent. eIF4G is a key target of picornaviral protease 2A, which cleaves this initiation factor into eIF4G(Delta) and (Delta)eIF4G to redirect the cellular translation machinery exclusively to its own IRES-containing transcripts. We have designed a novel translation control system (TCS) for conditional as well as adjustable translation of cap- and IRES-dependent transgene mRNAs in mammalian cells. eIF4G(Delta) and (Delta)eIF4G were fused C- and N-terminally to the FK506-binding protein (FKBP) and the FKBP-rapamycin binding domain (FRB) of the human FKBP-rapamycin-associated protein (FRAP), respectively. Rapamycin-induced heterodimerization of eIF4G(Delta)-FKBP and FRB (Delta)eIF4G fusion proteins reconstituted a functional chimeric elongation factor 4G in a dose-dependent manner. Rigorous quantitative expression analysis of cap- and IRES-dependent SEAP- (human placental secreted alkaline phosphatase) and luc- (Photinus pyralis luciferase) encoding reporter constructs confirmed adjustable translation control and revealed increased production of desired proteins in response to dimerization-induced heterologous eIF4G in Chinese hamster ovary (CHO-K1) cells. PMID- 12768628 TI - Asymmetric synthesis of L-homophenylalanine by equilibrium-shift using recombinant aromatic L-amino acid transaminase. AB - L-Homophenylalanine (L-HPA) was asymmetrically synthesized from 2-oxo-4 phenylbutyric acid (2-OPBA) and L-aspartate using a recombinant aromatic amino acid transaminase (AroAT). To screen microorganisms having such an L-specific AroAT with a relaxed substrate inhibition in the asymmetric synthesis of unnatural amino acids, enrichment cultures were performed in a minimal media containing 50 mM L-HPA as a sole nitrogen source. To reduce the intracellular background synthetic activity by amino acid pools in the cells, a two-step screening method was used. The putative AroAT (i.e., AroATEs) from the screened Enterobacter sp. BK2K-1 was cloned, sequenced, and overexpressed in E. coli cells. The activity of the overexpressed AroATEs was 314-fold higher than that of the wild-type cell. The substrate specificities of the enzyme and homology search revealed that the cloned transaminase is true AroAT. The AroATEs showed a substrate inhibition by 2-OPBA from 40 mM in the asymmetric synthesis, which made it difficult to perform batch asymmetric synthesis of L-HPA at high concentrations of 2-OPBA. To avoid the substrate inhibition by 2-OPBA, intermittent addition of the solid-state substrate was attempted to obtain a high concentration of L-HPA. By using the cell extract (75 U) obtained from the recombinant E. coli harboring the AroATEs gene, the asymmetric synthesis of L-HPA at 840 mM of 2-OPBA resulted in >94% of conversion yield and >99% ee of L-HPA of optical purity. Due to the low solubility (<2 mM) of L-HPA in the reaction buffer, synthesized L-HPA was continuously precipitated in the reaction media, which drives the reaction equilibrium towards the product formation. After full completion of the reaction, L-HPA of high purity (>99% ee) was easily recovered by simple pH shift of the reaction media. This method can permit very efficient asymmetric synthesis of other unnatural amino acids using a single transaminase reaction. PMID- 12768629 TI - Vibrating membrane filtration for recovery and concentration of insect killing nematodes. AB - New experimental data are reported that demonstrate the use of a novel vibrating membrane filter (VMF) for the combined recovery and concentration of two species of nematodes, S. feltiae and P. hermaphrodita, from mature liquid fermentation cultures. The disk membrane module had a working surface area of 0.2 m(2) and was operated at a constant flow rate of 0.2 m(3) h(-1). The recovery of the viable nematodes from the spent media and nonviable nematodes was assisted by an independently imposed oscillatory motion of the disk assembly, which produced an intense shear field at the membrane surface with calculated mean values on the order of 10(4) s(-1). Adult (nonviable) nematodes in the fermentation culture were preferentially dissolved in a detergent (sodium dodecylsulfate) and successfully separated from the juveniles using the VMF equipment. Permeate fluxes on the order of 15 to 30 L/m(2/)h were achieved for an operating transmembrane pressure of 800 mbar. Industrial-scale liquid fermentation for the manufacture of nematodes as biopesticides produces the viable nematode life stages in low-concentration suspension containing large quantities of spent media and other waste material. The VMF equipment provided a flexible operation for separation, cleaning, and concentration of viable nematodes from the fermentation broths. PMID- 12768630 TI - Ex vivo monitoring of protein production in baculovirus-infected Trichoplusia ni larvae with a GFP-specific optical probe. AB - Trichoplusia ni larvae were infected with baculoviruses containing genes for the expression of ultraviolet optimized green fluorescent protein (GFPuv) and several product proteins. A GFP-specific optical probe was used to both excite the green fluorescent protein (lambda(ex) = 385 nm), and subsequently monitor fluorescence emission (lambda(em) = 514 nm) from outside the infected larvae. The probe's photodetector was connected to a voltmeter, which was used to quantify the amount of GFPuv expressed in infected larvae. Voltage readings were significantly higher for infected vs. uninfected larvae and, by Western analysis, linear with the amount of GFPuv produced. In addition, the probe sensitivity and range were sufficient to delineate infection efficiency and recombinant protein production for model proteins, chloramphenicol acetyltransferase and human interleukin-2. This work represents a critical step in developing an automated process for the production of recombinant proteins in insect larvae. PMID- 12768631 TI - The new era of cervical cancer prevention: the end of the Pap smear? PMID- 12768632 TI - Cytoplasmic vacuolation, intracytoplasmic lumina, and DPAS staining in ductal carcinoma of the breast. AB - A retrospective study was carried out on 30 cases of histologically proven invasive ductal carcinoma of the breast with a prior fine-needle aspiration (FNA) cytology. On evaluating the May-Grunwald-Giemsa (MGG) FNA smears, cytoplasmic vacuolation was observed in 70% cases. Positivity with periodic acid Schiff positive, diastase-resistant (DPAS) staining was observed in 90% of cases. The chi(2) value on a McNemar test was 4.16. Thus, DPAS staining was significantly superior to MGG staining for picking up cytoplasmic vacuoles (P < 0.05). In 56.67% cases, DPAS staining showed an improvement in score as compared to MGG smears. This was highly significant (P < 0.001) on Wilcoxon matched-pairs signed ranks test. Applying the strict criteria of thick-walled cytoplasmic vacuoles with a central darkly stained dot, none of our cases revealed true intracytoplasmic lumina. Larger studies are required to establish a role for DPAS staining in separating borderline, in situ, and invasive breast lesions, and to see if such positvity can be incorporated into the grading systems for breast carcinoma. PMID- 12768633 TI - Melanoma of the soft parts: diagnosis of metastatic and recurrent tumors by aspiration cytology. AB - Three cases of melanoma of the soft parts diagnosed by aspiration cytology and immunochemistry are described. Unusual clinicoradiological features, variable cytomorphology, absence of melanin, and lack of awareness due to rarity of the lesion caused diagnostic dilemma. Morphological similarity to alveolar soft part sarcoma, synovial and epithelioid sarcoma, cutaneous melanoma, poorly differentiated carcinoma, and plasmacytoma were noted. Consideration of clinical data such as age, site, and radiology was helpful in narrowing down the diagnosis. A presumptive cytologic diagnosis helped in selecting the antibody panel, which established the definitive diagnosis in all cases. This approach is valuable in preoperative management and obviates the need for an open biopsy in primary, metastatic, or recurrent tumor. Increasing experience in fine-needle aspiration of this rare entity and correlation with immunochemistry and histopathology are valuable in narrowing down the differential diagnosis, reducing diagnostic errors, and widening its clinicocytological spectrum. PMID- 12768635 TI - Atypical squamous cells of undetermined significance--rule out high-grade squamous intraepithelial lesion: cytopathologic characteristics and clinical correlates. AB - The significance and clinical management of atypical squamous cells of undetermined significance (ASCUS) on cervical cytologic smears has been an area of much controversy. This study compiled a list of criteria useful in identifying the subset of cases that would be categorized as atypical squamous cells of undetermined significance- rule out high-grade squamous intraepithelial lesion (ASC-H) in the new Bethesda System terminology, which eventuate in a diagnosis of cervical intraepithelial neoplasia (CIN). The cytopathology files at Johns Hopkins Hospital were searched for ASC-H cases from the 3-yr period 1996-1998, which had definitive clinicopathologic follow-up (colposcopy and cervical biopsies). The smears were reviewed, cytomorphologic features studied, and clinical correlations performed. ASC-H was diagnosed in 257 of 45,428 gynecologic smears (0.6%), 72 having had clinicopathologic follow-up. Of these 72 cases, 35 (49%) on follow-up had a negative/reactive diagnosis (NR), whereas 37 (51%) turned out to be CIN [CIN-I-18 (49%) and CIN II and III-19 (51%)]. The significant cytomorphologic differences in the ASC-H category with a CIN follow up (compared with an NR follow-up) were fewer atypical cells, more often discohesive or seen singly, more monomorphic, a higher nuclear-to-cytoplasmic (N/C) ratio, greater nuclear hyperchromasia, more coarse, unevenly dispersed chromatin, more prominent nuclear membrane irregularities, lack of nucleoli, chromocenters or nuclear grooves, and lack of an inflammatory background. Careful attention to subtle cytomorphologic characteristics may be helpful for a more definitive subdivision of ASC-H terminology into a NR and a CIN diagnosis. PMID- 12768634 TI - Evaluation of fluorescence in situ hybridization as an ancillary tool to urine cytology in diagnosing urothelial carcinoma. AB - Our purpose was to evaluate the feasibility of performing fluorescence in situ hybridization (FISH) on routine urine samples and to compare the relative sensitivities of urine cytology and FISH for detecting urothelial carcinoma. Light microscopy (LM) using cytologic evaluation and FISH were used to study 121 consecutive urine samples. A mixture of fluorescent probes to chromosomes 3, 7, 17, and the 9p21 locus were used for detection of numerical chromosomal abnormalities (UroVysion, Vysis/Abbott). Biopsy specimens from patients in the study were reviewed if available. FISH analysis was performed without knowledge of cytology or biopsy findings. The urine cytology of 121 samples was interpreted as 59 negative, 41 reactive, 16 atypical, 2 suspicious and 3 insufficient cells for diagnosis. 85 samples were successfully analyzed by FISH. Thirty-one of these showed chromosomal abnormalities and these samples were initially regarded on the original cytology reading as follows: 10 negative, 10 reactive, 9 atypical, and 2 suspicious. FISH demonstrated chromosomal abnormalities in a significant number of cases (67%) that were initially diagnosed as normal or reactive by LM. Twenty five patients were identified who had biopsy-proven TCC and successful FISH. Thirteen of the 25 patients (52%) were abnormal by FISH (cytology: 2 suspicious, 6 atypical, 4 reactive, 1 negative). One patient was atypical by cytology with normal FISH results but had TCC on biopsy. Hyperdiploidy for chromosomes 3 (77%) and 7 (67%) were seen consistently. Multiple chromosomal abnormalities were seen in 67% of these cases. We conclude that FISH has a greater sensitivity in detecting urothelial carcinoma when coupled with urine cytology. It is not entirely clear at this time whether a positive FISH may indicate frank neoplastic urothelial transformation or merely be an indicator of unstable urothelium capable of or primed for malignant transformation thus detecting patients at significant risk. The use of FISH in conjunction with urine cytology can potentially reduce urothelial carcinoma morbidity and mortality by diagnosing these tumors earlier. PMID- 12768636 TI - Clear cell sarcoma (malignant melanoma of soft parts): fine-needle aspiration cytology of a highly pigmented tumor. AB - Diagnosis of clear cell sarcoma by fine-needle aspiration is difficult, as melanin is usually absent. Here we describe a case of a primary clear cell carcinoma with abundant melanin, making diagnosis definitive. PMID- 12768638 TI - Fine-needle aspiration of rheumatoid nodule: a case report with review of diagnostic features and difficulties. AB - Patients with rheumatoid arthritis may develop extra-articular subcutaneous nodules as part of the systemic disease or as initial manifestation. These lesions may represent significant diagnostic dilemmas in patients with clinical suspicion of malignancy. In this setting, fine-needle aspiration (FNA) of the nodules may be the simplest and most appropriate diagnostic approach. In the literature, however, there are only sporadic reports describing FNA cytology of a rheumatoid nodule. In this report, we present a case of a 67-year-old male with a history of rheumatoid arthritis and squamous-cell carcinoma of the larynx who developed a subcutaneous neck nodule in the immediate proximity of the surgical scar. Clinically, because of the history of squamous-cell carcinoma and location of the lesion, the nodule was suspected to be metastatic cancer, but was proven by FNA biopsy to represent rheumatoid disease. Cytological criteria of rheumatoid nodule and diagnostic difficulties are discussed. PMID- 12768637 TI - Glomus tumor of the stomach: cytologic diagnosis by endoscopic ultrasound-guided fine-needle aspiration. AB - The glomus tumor is usually a benign solitary neoplasm that arises from modified smooth muscle cells of the glomus body, a type of neuromyoarterial receptor that plays a role in the regulation of arterial blood flow. We report a case of gastric glomus tumor diagnosed by endoscopic ultrasound-guided fine-needle aspiration. Smears exhibited groups of cohesive, uniform, small, round to polygonal cells with scant cytoplasm, indistinct cell borders, and round, hyperchromatic nuclei with homogeneous chromatin. The cell block contained fragments of cells with similar morphologic features and immunohistochemical staining exhibited positivity for smooth muscle actin and vimentin. The diagnosis was confirmed by the surgically resected specimen. Ultrastructural examination revealed prominent pinocytotic vesicles lining the plasmalemma. In this report, we discuss the differential diagnosis of gastric glomus tumor and compare the cytologic features of this case with two others reported in the literature. PMID- 12768639 TI - Amyloid tumor: a clinical and cytomorphologic study. AB - We describe the cytologic findings and clinical presentation of three unusual cases of amyloid tumor. Two of our patients had low-grade lymphoid malignancies and the third insulin-dependent diabetes mellitus. In no cases was amyloid suspected as the cause of mass lesion. Two of our cases presented with superficial soft tissue mass and the third with right breast masses and bilateral axillary lymph node enlargement. Air-dried slides from all aspirated cases were stained with Diff-Quik for specimen adequacy evaluation. The remaining fixed slides were stained with Papanicolaou stain. Amyloid appeared as dark-blue to purple clumps of acellular material on Diff-Quik stain, accompanied with chronic inflammatory cell infiltrates and multinucleated giant cells, simulating granulomatous inflammation. Papanicolaou stain demonstrated cyanophilic to orangophilic acellular material. Amyloid was suspected and subsequently confirmed by Congo red stain. PMID- 12768640 TI - Is core needle biopsy superior to fine-needle aspiration biopsy in the diagnosis of papillary breast lesions? AB - Since the 1980s core needle biopsy (CNB) has gained remarkable popularity and in many institutions it has replaced fine-needle aspiration biopsy (FNAB). However, similar to FNAB, limitation remains in the ability of this procedure to reliably diagnose a small, but prognostically significant, number of breast lesions. These include entities such as atypical ductal hyperplasia, fibro-epithelial tumors, radial scar, papillary lesions, and lobular neoplasia. To assess the diagnostic accuracy of CNB vs. FNAB in the same breast lesions, we reviewed our cases of papillary lesions of the breast. In a retrospective study, we identified 36 cases of FNAB and 11 cases of CNB diagnosed as papillary lesions and compared the results with their corresponding surgical specimen. Interpretation ranged from papillary vs. atypical papillary lesions favoring benign vs. malignant tumors, respectively. Occasionally, definitive diagnosis of papillary carcinoma was entertained. Immunohistochemical staining with smooth muscle actin was used to evaluate the presence or absence of a myoepithelial cell layer. FNAB had benign findings in 21 lesions, atypical in 10, and malignant in five. Of the five lesions yielding malignant features, four had invasive carcinoma and one had micropapillary ductal carcinoma in situ (DCIS). Surgery revealed invasive carcinoma in three of the cases interpreted as atypical papillary lesions and invasive cancer and micropapillary DCIS in three of the cases diagnosed as benign lesions. Similar results were obtained with CNB. DCIS was found in one out of six of the cases diagnosed as papilloma. Out of the four cases that were interpreted as atypical papillary lesion, surgery revealed invasive carcinoma in one case and one case had micropapillary DCIS. Diagnosis of malignancy was confirmed by histology in one case interpreted as papillary carcinoma by CNB. This study suggested that both FNAB and CNB share similar diagnostic challenges and a follow up surgical excision is indicated when diagnosis of a papillary lesion is entertained by both procedures. PMID- 12768641 TI - Diagnostic pitfalls of peritoneal washing cytology and the role of cell blocks in their diagnosis. AB - Mesothelial cell hyperplasia, collagen balls, endometriosis, and endosalpingiosis are diagnostic pitfalls on peritoneal washing cytology in women who present with gynecologic lesions. Over an 8-month period, the peritoneal washings from 10 patients undergoing gynecologic surgery for presumed malignancy showed unusual cytologic findings, several of which posed diagnostic difficulties. The washings from four patients with ovarian carcinomas were cellular and contained clusters and strips of cells with cytologic atypia mimicking malignancy. Confirmation of their benign mesothelial origin was confirmed on immunohistochemistry utilizing cell block preparations. In two cases of endometrial endometrioid carcinoma, the washings contained several clusters of cells surrounding and/or admixed with a globular substance. Due to their similarity to endometrial cells, immunohistochemistry was performed on cell block preparations. The cells were positive for cytokeratin and negative for carcinoembryonic antigen and B72.3, confirming their mesothelial origin. In one case, clinically presumed to be a malignant mass, the washings contained tight clusters of cells with mild cytologic atypia admixed with hemosiderin-laden macrophages. In conjunction with the cell block findings, a diagnosis of endometriosis was made. Extensive endometriosis was found on the surgically resected specimen. In two cases, strips of ciliated epithelial cells resembling tubal epithelium were present on the cytologic and cell block preparations, consistent with endosalpingiosis. The peritoneal washings in one case contained several clusters and balls of atypical cells surrounding microcalcifications on cell block preparation. Since calcification within groups of cells in peritoneal washings always raised the possibility of malignancy, a serous carcinoma of the ovary, particularly of borderline malignancy, would have to be excluded. Fortunately, the resected specimen was free of tumor and showed calcified endosalpingiosis on the ovarian surface. Preparation of cell blocks from peritoneal washings is of value in the work-up and management of patients who present with cytologic mimickers of malignancy on fluid cytology. PMID- 12768642 TI - A case of omental gastrointestinal stromal tumor and association with history of melanoma. PMID- 12768643 TI - Fine-needle aspiration cytology: its origin, development, and present status with special reference to a developing country, India. AB - Fine-needle aspiration cytology (FNAC) was performed on a large scale at Memorial Hospital, New York, during the 1930s, but during the ensuing years, it did not gain much encouragement in United States. The technique had a resurgence in Scandinavia during the 1950s and 1960s, where it flourished before spreading to other parts of the world. It had also a revival in the United States, which contributed enormously to this tool in each and every aspect. The status of FNA during 1966-2002 was assessed through review of MEDLINE search data on FNA and its correlation with World Bank website data on classification of countries. A total of 849 journals published 5,609 articles on FNA over a period of 37 years. Both the number of publishing journals and the number of published articles on FNA were low during the 1960s (3.5 +/- 0.58 and 4.0 +/- 0.82, respectively) and 1970s (20.3 +/- 14.72 and 25.0 +/- 20.54, respectively), but their number increased sharply from the 1980s onward (78.2 +/- 25.65 and 147.2 +/- 66.89, respectively, during the 1980s, 126.2 +/- 11.94 and 301.4 +/- 35.99, respectively, during the 1990s, and 113.3 +/- 36.46 and 287.3 +/- 85.93, respectively, during the 2000s). The difference between the decades of 1960s 2000s, with respect to the number of publishing journals and published articles, was highly significant (P < 0.0001). Only 90 (10.6%) of the journals were from the arena of pathology and its branches. The remaining journals belonged to various other disciplines of medicine; a small fraction were even from the veterinary sciences. Ten journals, including three in the field of cytopathology, published 2,448 (43.6%) of the total articles on FNA. During 1987-2002, 46 (29.7%) of the 155 developing nations published articles on FNA, whereas 28 (52.8%) of the developed (high-income economies) countries did so, the difference being highly significant (P = 0.0044). The total number of publications from high income economies was 3,124 (195.3 per year), as opposed to 772 (48.3 per year) from the developing world. The number of articles published from the developing nations (16.8 +/- 52.21) was significantly lower as compared with that from the high-income economies (111.6 +/- 242.03) (P = 0.005). Except for infectious diseases, the high-income economies had a definite edge over the developing nations in the absolute number of publications from each and every site/organ. However, when the frequency of publications on various organs/sites was compared between the two groups, it was found that the number of articles from high-income economies was significantly higher with respect to breast diseases (P < 0.001) and pancreatic lesions (P = 0.0158), whereas the developing nations published more frequently on small round cell tumors (excluding exclusive reports on lymphoma) and infectious diseases (P < 0.001). In India, FNAC was first introduced during the early 1970s and spread to different parts through formal teaching under the postgraduate curriculum in pathology and by conducting workshops and continuing medical education program for pathologists, surgeons/physicians, and radiologists. FNAC is now practiced in all medical colleges, in big public sector hospitals, and even in private clinics and laboratories. The number of centers practicing FNAC increased sharply during 1980s, as evident from the response of 69 laboratories in various parts of India to a questionnaire. As of 1998, 55.9% of the laboratories performed >/=1,000 cases of FNA per year. In 46% of the centers, pathologists alone performed the FNAC, whereas in 51%, they performed it in collaboration with radiologists and surgeons. Disposable syringes and needles were used in all the centers, but syringe holders were used in only in 61% of centers. In 86% of laboratories, two or more routine stains were used, and one or more ancillary techniques on fine needle aspirates were adopted in 72.5% centers. Of the 772 publications from the developing world during 1987-2002, India alone contributed to 374 (48.4%). During 1975-2002, 154 articles on FNA were published needles were used in all the centers, but syringe holders were used in only in 61% of centers. In 86% of laboratories, two or more routine stains were used, and one or more ancillary techniques on fine-needle aspirates were adopted in 72.5% centers. Of the 772 publications from the developing world during 1987-2002, India alone contributed to 374 (48.4%). During 1975-2002, 154 articles on FNA were published in 15 indexed Indian journals. PMID- 12768644 TI - Anxiety disorders in women: a developmental and lifecycle perspective. PMID- 12768645 TI - Brain mechanisms underlying emotional alterations in the peripartum period in rats. AB - In the period before and after parturition, i.e., in pregnancy and lactation, a variety of neuroendocrine alterations occur that are accompanied by marked behavioral changes, including emotional responsiveness to external challenging situations. On the one hand, activation of neuroendocrine systems (oxytocin, prolactin) ensures reproduction-related physiological processes, but in a synergistic manner also ensures accompanying behaviors necessary for the survival of the offspring. On the other hand, there is a dramatic reduction in the responsiveness of neuroendocrine systems to stimuli not relevant for reproduction, such as the hypothalamo-pituitary-adrenal (HPA) axis responses to physical or emotional stimuli in both pregnant and lactating rats. With CRH being the main regulator of the HPA axis, downregulation of the brain CRH system may result in various behavioral, in particular emotional, adaptations of the maternal organisms, including changes in anxiety-related behavior. In support of this, the lactating rat becomes less emotionally responsive to novel situations, demonstrating reduced anxiety, and shows a higher degree of aggressive behavior in the test for agonistic behavior as well as in the maternal defense test. These changes in emotionality are independent of the innate (pre-lactation) level of anxiety and are seen in both rats bred for high as well as low levels of anxiety. Both brain oxytocin and prolactin, highly activated at this time, play a significant role in these behavioral and possibly also neuroendocrine adaptations in the peripartum period. PMID- 12768646 TI - How does premenstrual dysphoric disorder relate to depression and anxiety disorders? AB - Premenstrual dysphoric disorder (PMDD) is a severe variant of premenstrual syndrome that afflicts approximately 5% of all women of fertile age. The hallmark of this condition is the surfacing of symptoms during the luteal phase of the menstrual cycle, and the disappearance of symptoms shortly after the onset of menstruation. Whereas many researchers have emphasized the similarities between PMDD and anxiety disorders, and in particular panic disorder, others have suggested that PMDD should be regarded as a variant of depression. Supporting both these notions, the treatment of choice for PMDD, the serotonin reuptake inhibitors (SRIs), is also first line of treatment for depression and for most anxiety disorders. In this review, the relationship between PMDD on the one hand, and anxiety and depression on the other, is being discussed. Our conclusion is that PMDD is neither a variant of depression nor an anxiety disorder, but a distinct diagnostic entity, with irritability and affect lability rather than depressed mood or anxiety as most characteristic features. The clinical profile of SRIs when used for PMDD, including a short onset of action, suggests that this effect is mediated by other serotonergic synapses than the antidepressant and anti-anxiety effects of these drugs. Although we hence suggest that PMDD should be regarded as a distinct entity, it should be emphasized that this disorder does display intriguing similarities with other conditions, and in particular with panic disorder, which should be the subject of further studies. Also, the possibility that there are subtypes of PMDD more closely related to depression, or anxiety disorders, than the most common form of the syndrome, should not be excluded. PMID- 12768647 TI - The weaker sex? Gender and post-traumatic stress disorder. AB - We discuss the gender-specific differences for traumatic events and Post Traumatic Stress Disorder (PTSD) as found in the epidemiological literature. Recent research literature consistently reports three interesting findings: 1) men experience traumatic events more often, 2) women and men differ in the type of traumatic experiences they experience, and 3) women more often develop PTSD after the experience of a traumatic event. In the second part of the present article we provide some explanations for these differences. The reported higher vulnerability of women for PTSD could be due to the methodology used, the higher prevalence of childhood sexual abuse and rape in women, the different coping styles of women and men, or the more limited socio-economic resources of women. PMID- 12768648 TI - Pregnancy anxiety and comorbid depression and anger: effects on the fetus and neonate. AB - One hundred sixty-six women were classified as experiencing high or low anxiety during the second trimester of pregnancy. The high anxiety women also had high scores on depression and anger scales. In a follow-up across pregnancy, the fetuses of the high anxiety women were noted to be more active and to experience growth delays. The high anxiety mothers' high prenatal norepinephrine and low dopamine levels were followed by their neonates having low dopamine and serotonin levels. The high anxiety mothers' newborns also had greater relative right frontal EEG activation and lower vagal tone. Finally, the newborns of high anxiety mothers spent more time in deep sleep and less time in quiet and active alert states and showed more state changes and less optimal performance on the Brazelton Neonatal Behavior Assessment Scale (motor maturity, autonomic stability and withdrawal). These data highlight the need for prenatal intervention for elevated anxiety symptoms during pregnancy. PMID- 12768649 TI - Depressive symptoms at two months after miscarriage: interpreting study findings from an epidemiological versus clinical perspective. AB - We identify two study designs used to investigate psychiatric symptoms following reproductive loss. One design examines psychiatric outcomes in a single cohort of bereaved women; the other compares psychiatric outcomes in bereaved and non bereaved cohorts. We employ the findings from one study of depressive symptoms following miscarriage to exemplify these designs and inferential differences in causal inferences that can be drawn. We interviewed a cohort of 114 women at 6-8 weeks after miscarriage and a cohort of 318 community women not recently pregnant. Depressive symptoms were assessed with the Center for Epidemiologic Studies Depression (CES-D) Scale. Among miscarrying women, the mean level of depressive symptoms was substantially elevated above that in the community cohort (adjusted difference in means between cohorts, 4.9, 95% confidence interval [CI] 2.3-7.4). Among miscarrying women, 20.2% were highly symptomatic (CES-D 30+), as compared with 10.1% among community women (adjusted odds ratio [OR] 2.8, 95% CI 1.4-5.6). However, in contrast to findings from the same study concerning depressive reactions at 2 weeks after loss, the impact of miscarriage on depressive symptoms at 6-8 weeks was markedly greater for younger women and possibly for women with a history of prior reproductive loss, but otherwise did not vary with number of living children, marital status, ethnicity or educational level. We conclude that miscarrying women first seen at their 6- to 8-week post miscarriage gynecologic visit are likely to be more depressed on average than otherwise comparable women who have not experienced a recent reproductive loss. Factors that moderate the impact of miscarriage may vary with time since loss. Enhanced recognition of the implications of study design for research inferences may help bring greater clarify and uniformity to findings from future investigations in this field. PMID- 12768650 TI - Are there gender differences in objective and subjective sleep measures? A study of insomniacs and healthy controls. AB - It is well known that insomnia is more frequent in women than in men throughout all age groups. In this respect insomnia resembles other psychiatric disorders that occur more frequently in women such as anxiety and depressive disorders. Since insomnia is frequently a symptom of anxiety and depression, it remains an open question whether the comorbidity with psychiatric disorders fully explains the gender differences in the prevalence of insomnia or whether gender influences sleep independently from psychiatric conditions. We analyzed sleep measures of patients diagnosed with a primary insomnia (n=86) and of an age- and sex-matched healthy control group (n=86) by polysomnography; additionally, subjective rating scales were available for 70 patients and 54 controls matched for mean age and sex ratio. Surprisingly, none of the sleep continuity measures (sleep duration, sleep efficiency, arousal index, and wake%), nor slow wave or REM sleep % showed significant gender differences in both insomniacs and healthy controls. Also, subjective estimates of sleep quality were comparable in both sexes. As expected, insomniacs strongly differed from the control group in all subjective measures of sleep. Polysomnography showed significantly reduced sleep duration and efficiency, increased arousal index, and slightly, but significantly, less REM sleep in the insomniacs as compared to the healthy controls. These studies indicate that gender seems to have, if any, relatively little influence on sleep per se. We hypothesize that the clear gender differences in the prevalence of insomnia are caused predominantly by gender differences in the prevalence of anxiety and depression. Primary insomnia may be, at least in a part of the cases, a subclinical or subthreshold form of anxiety or depression. PMID- 12768651 TI - Chronicity, relapse, and illness--course of panic disorder, social phobia, and generalized anxiety disorder: findings in men and women from 8 years of follow up. AB - Anxiety disorders are chronic illnesses that occur more often in women than men. Previously, we found a significant sex difference in the 5-year clinical course of uncomplicated panic disorder that was attributable to a doubling of the illness relapse rate in women compared to men. However, we have not detected a sex difference in the clinical course of panic with agoraphobia, generalized anxiety disorder (GAD), or social phobia (SP), which are conditions generally thought to be more chronic than uncomplicated panic disorder. Given that a longer follow-up period may be required to detect differences in clinical course for more enduring illnesses, we conducted further analyses on this same cohort after a more protracted interval of observation to determine whether sex differences would emerge or be sustained. Data were analyzed from the Harvard/Brown Anxiety Research Program (HARP), a naturalistic, longitudinal study that repeatedly assessed patients at 6 to 12 month intervals over the course of 8 years. Data regarding remission and relapse status were collected from 558 patients and treatment was observed but not prescribed. Cumulative remission rates were equivalent among men and women with all diagnoses. Patients who experienced remission were more likely to improve during the first 2 years of study. Women with GAD continued remitting late into the observation period and experienced fewer overall remission events by 8 years. However, the difference in course failed to reach statistical significance. Relapse rates for women were comparable to those for men who suffered from panic disorder with agoraphobia, GAD, and SP. Again, initial relapse events were more likely to occur within the first 2 years of observation. However, relapse events for uncomplicated panic in women were less restricted to the first 2 years of observation and by 8 years, the relapse rates for uncomplicated panic was 3-fold higher in women compared with men. Anxiety disorders are chronic in the majority of men and women, although uncomplicated panic is characterized by frequent remission and relapse events. Short interval follow-up shows sex differences in the remission and relapse rates for some but not all anxiety disorders. These findings suggest important differences in the clinical course among the various anxiety disorders and support nosological distinctions among the various types of anxiety. It may be that sex differences in the clinical course of anxiety disorders hold prognostic implications for patients with these illnesses. PMID- 12768652 TI - Stresses on women physicians: consequences and coping techniques. AB - We review current data on types of stressors acting on women physicians, the consequences of these stressors and methods of coping with them. We undertook a systematic review of original articles published in the last 15 years and registered mainly on Medline and on the internet websites focusing on these issues. In addition to the pressures acting on all physicians, women physicians face specific stressors related to discrimination, lack of role models and support, role strain, and overload. The depression rate in women physicians does not vary from that of the general public but the rates of successful suicide and divorce are much higher. Women in academic settings are promoted more slowly, have lower salaries, receive fewer resources, and suffer from a range of micro inequities. They often lack mentors to provide advice and guidance. They must cope with the pressures of choosing when to have a child and conflicts between being a wife and mother and having a career. Despite these pressures, they report a high degree of career satisfaction. Although women physicians suffer from a variety of stressors that can lead to career impediments, stress reactions, and psychiatric problems, generally they are satisfied with their careers. Personal coping techniques can help women deal with these stressors. Pressures will continue until attitudes and practices change in institutional settings. Some institutions are initiating changes to end discrimination against women faculty. PMID- 12768653 TI - Neural crest and cardiovascular development: a 20-year perspective. AB - BACKGROUND: Twenty years ago this year was the first publication describing a region of neural crest cells necessary for normal cardiovascular development. Ablation of this region in chick resulted in persistent truncus arteriosus, mispatterning of the great vessels, outflow malalignments, and hypoplasia or aplasia of the pharyngeal glands. METHODS: We begin with a historical perspective and then review the progress that has been made in the ensuing 20 years in determining the direct and indirect contributions of the neural crest cells, now termed cardiac neural crest cells, in cardiovascular and pharyngeal arch development. Many of the molecular pathways that are now known to influence the specification, migration, patterning and final targeting of the cardiac neural crest cells are also reviewed. RESULTS: Although much knowledge has been gained by using many genetic manipulations to understand the cardiac neural crest cells' role in cardiovascular development, most models fail to explain the phenotypes seen in syndromic and non-syndromic human congenital heart defects, such as the DiGeorge syndrome. CONCLUSIONS: We propose that the cardiac neural crest exists as part of a larger cardiocraniofacial morphogenetic field and describe several human syndromes that result from abnormal development of this field. PMID- 12768655 TI - T-box genes and cardiac development. AB - BACKGROUND: T-box genes play roles in vertebrate gastrulation and in later organogenesis. Their existence in all metazoans examined so far indicates that this is an evolutionarily ancient gene family. Drosophila melanogaster has eight T-box genes, whereas Caenorhabditis elegans has 22. Mammals appear to have at least 18 T-box genes, comprising five subfamilies. METHODS: A full range of cytological, developmental, molecular and genetic methodologies have recently been applied to the study of T-box genes. RESULTS: Over the last 5 years, mutations in TBX1 and TBX5 have been implicated in two human disorders with haplo insufficient cardiovascular phenotypes, DiGeorge/velocardiofacial syndrome and Holt-Oram ("heart-hand") syndrome. Interestingly, the number of T-box gene family members discovered to have cardiac or pharyngeal arch expression domains during vertebrate embryonic development has steadily grown. In addition, various Tbx5 loss-of-function models in organisms as distant as the mouse and zebrafish do indeed phenocopy Holt-Oram syndrome. Finally, the intriguing discovery earlier this year that a T-box gene is expressed in a subset of cardioblasts in D. melanogaster suggests that members of this gene family may have fundamental, conserved roles in cardiovascular pattern formation. CONCLUSIONS: These developments prompted us to review the current understanding of the contribution of T-box genes to cardiovascular morphogenesis. PMID- 12768654 TI - Regulation of heart morphology: current molecular and cellular perspectives on the coordinated emergence of cardiac form and function. AB - BACKGROUND: During early heart development, in addition to cells being induced to differentiate into cardiomyocytes, pathways are activated that lead to cardiac morphogenesis or the development of form. METHODS: Orchestration of organogenesis involves the incremental activation of regulatory pathways that lead to pivotal transition points, such as cardiac compartment delineation and looping. Each embryonic stage sets up the correct patterning of morphoregulatory molecules that will regulate the next process, until an organ is formed from the mesoderm layer after gastrulation. The current review provides an understanding of the morphoregulatory, cell adhesion and extracellular matrix-mediated, processes that coordinate development of heart form with that of function. The period reviewed encompasses the formation of a definitive cardiac compartment from the lateral plate mesoderm to the time-point in which the single, beating heart tube loops directionally to the right. Looping results in the correct spatial orientation for subsequent modeling of the four-chambered heart. Even subtle alterations in looping can form the basis upon which malformations of the inlet or the outlet regions of the heart, or both, are superimposed. RESULTS: In the future, DNA microarray data sets may allow modeling the specific sequence of gene regulatory dynamics leading to these transition points to discover the regulatory "modes" that the cells adopt during heart organogenesis. The regulatory genes, however, can only specify the proteins that will be present. CONCLUSIONS: To fully understand the timing and mechanisms underlying heart development, it is necessary to define the sequential synthesis, patterning, and interaction of the proteins, and of still other receptors, which eventually drive cells to organize into functioning organs. PMID- 12768656 TI - Sculpting the cardiac outflow tract. AB - The cardiac outflow tract is the site of anomalies that affect a substantial proportion of individuals with congenital heart defects. The morphogenesis of this site is complex, and requires coordinated development of many cell types and tissues. It is therefore not surprising that developmental mistakes arise here, and that the steps and mechanisms of morphogenesis are still controversial and poorly understood, despite advances in molecular techniques. Recent findings have provided new insight into mechanisms of outflow tract morphogenesis, including clarification of its origins and the fate of cardiomyocytes, as well as invading cell populations. Application of new and old techniques and a wide range of approaches to tackle the unanswered questions about the outflow tract calls for collaboration among investigators from different disciplines including anatomists, physiologists, and molecular biologists. PMID- 12768657 TI - Development of the cardiac pacemaking and conduction system. AB - The heartbeat is initiated and coordinated by a heterogeneous set of tissues, collectively referred to as the pacemaking and conduction system (PCS). While the structural and physiological properties of these specialized tissues has been studied for more than a century, distinct new insights have emerged in recent years. The tools of molecular biology and the lessons of modern embryology are beginning to uncover the mechanisms governing induction, patterning and developmental integration of the PCS. In particular, significant advances have been made in understanding the developmental biology of the fast conduction network in the ventricles--the His-Purkinje system. Although this progress has largely been made by using animal models such as the chick and mouse, the insights gained may help explain cardiac disease in humans, as well as lead to new treatment strategies. PMID- 12768658 TI - Early events in valvulogenesis: a signaling perspective. AB - The proper formation and function of the vertebrate heart requires a multitude of specific cell and tissue interactions. These interactions drive the early specification and assembly of components of the cardiovascular system that lead to a functioning system before the attainment of the definitive cardiac and vascular structures seen in the adult. Many of these adult structures are hypothesized to require both proper molecular and physical cues to form correctly. Unlike any other organ system in the embryo, the cardiovascular system requires concurrent function and formation for the embryo to survive. An example of this complex interaction between molecular and physical cues is the formation of the valves of the heart. Both molecular cues that regulate cell transformation, migration, and extracellular matrix deposition, and physical cues emanating from the beating heart, as well as hemodynamic forces, are required for valvulogenesis. This review will focus on molecules and emerging pathways that guide early events in valvulogenesis. PMID- 12768659 TI - Embryonic and adult vasculogenesis. AB - Two mechanisms account for the formation of blood vessels, vasculogenesis and angiogenesis. Unfortunately, the terms vasculogenesis and angiogenesis literally have the same meaning, i.e., the genesis of blood vessels, and thus do little to distinguish between the two processes. Despite the nomenclature, the two processes are clearly distinct. Vasculogenesis, the de novo formation of blood vessels from mesoderm, is driven by the recruitment of undifferentiated mesodermal cells to the endothelial lineage and the de novo assembly of such cells into blood vessels. Angiogenesis is the generation of new blood vessels from endothelial cells of existing blood vessels, a process driven by endothelial cell proliferation. Recent years have seen dramatic changes in our understanding of the process of vasculogenesis, expanding the scope of its occurrence beyond the earliest stages of development to include involvement in neovascular processes throughout development as well as in the adult. In this review, emphasis is placed on discussion of emerging perspectives on the process of vasculogenesis in both the embryo and the adult. PMID- 12768660 TI - Noninvasive phenotypic analysis of cardiovascular structure and function in fetal mice using ultrasound. AB - METHODS: We established methods for noninvasive mouse fetal heart imaging using an Acuson/Siemens Sequoia ultrasound scanner equipped with a single-pulse CHIRP Coded Excitation program, and a highfrequency linear array transducer. Mouse fetuses spanning gestation day 12.5 to 18.5 (E12.5-E18.5) were studied. RESULTS: Controlled anesthetic and constant body temperature were found to be essential for hemodynamic stability of the mother and fetuses. Fetal heart rates increased from 160 to 220 beats/min as development progressed. These heart rates were lower than those of newborn mice, so that frame rates above 100 Hz adequately resolved structural details in 2D without misregistration. Analysis of 2D images showed a doubling in crown-torump length (8-19 mm), and rapid growth of the heart from 1 to 3 mm in diameter as fetuses developed from E12.5 to E18.5. A cumulative increase in scanning modalities was achieved with increasing developmental age, with the optimal stage for scanning being E16.5. At E16.5 right and left could be distinguished, and it was possible to obtain diagnostic 2D color flow Doppler in the four-chamber, apical long axis 3/5-chamber and short axis views. In addition, M-mode images of high quality were obtainable from E15.5 to E18.5, whereas spectral Doppler signals could be obtained readily from E12.5 onwards. CONCLUSIONS: These studies show that ultrasound imaging can be used for structural and functional analysis of the developing mammalian heart, even at early stages of development. Such noninvasive cardiovascular ultrasonic evaluation should be ideally suited for high throughput screening of mutagenized mice. PMID- 12768661 TI - [The role of mtDNA deletion in the sensitivity to aminoglycoside antibiotic induced deafness]. AB - OBJECTIVE: To establish an animal model of mitochondrial DNA (mtDNA) deletion and investigate the possible role of mtDNA deletion in aminoglycoside antibiotic induced deafness. METHODS: Thirty wistar rats (4 months) were randomly divided into group A and B. Doxorubicin (DOX) was subcutaneously injected at doses of 2 mg/kg twice per week for 3 months in group A and then kanamycin (KM) was intraperitoneally injected 500 mg/kg per day for 10 consecutive days. The treatments of group B were identical to group A, except normal saline was substituted for DOX. The thresholds of auditory brainstem response (ABR) were measured before and after the drug administrations. The inner ear membranous labyrinthine tissue was harvested and mtDNA was amplified to identify 4,834 bp deletion by PCR technique. RESULTS: The elevation of the mean ABR thresholds in group A(67.08 +/- 8.59) dB peSPL was significantly higher than that in group B (12.71 +/- 4.42) dB peSPL after KM administration (P < 0.001). In group A, 9 of the 15 rats demonstrated 4,834 bp mtDNA deletion. However, mtDNA 4,834 bp deletion was negative in group B animals. CONCLUSION: DOX can induce mtDNA deletion in the inner ear tissue of the rat. mtDNA deletion in the inner ear may play an important role in the hypersensitivity to aminoglycoside antibiotic ototoxicity. PMID- 12768662 TI - [Audiological findings and mitochondrial DNA mutation in a large family with matrilineal sensorineural hearing loss]. AB - OBJECTIVE: To explore audiological features of matrilineal non-syndromic deafness and its molecular mechanism. METHODS: A large family with 41 members having inherited deafness was studied. Complete history and the data of general and otolaryngological examinations were collected. All subjects were screened for mitochondrial DNA A1555G mutation by molecular analysis. Audiological evaluation included puretone audiometry, auditory brainstem responses and transiently evoked otoacoustic emissions. RESULTS: All subjects were in good health generally. Molecular analysis showed that all maternal relatives with or without hearing loss harbored the A1555G mitochondrial mutation. No mutation was found among spouses and paternal relatives. Audiological results showed notable symmetric bilateral sensorineural hearing loss in 17 of 20 maternal relatives, in which 5 cases had a progressive hearing loss in the recent 11 years. The age of appearance of hearing loss ranged from 1 to 50 years. CONCLUSION: All hearing impaired subjects of this family had late-onset sensorineural hearing loss. Most of which were progressive. The A1555G mitochondrial mutation in the 12S rRNA gene is responsible for the disorder. Other factors, such as nuclear genes or environmental determinants, may influence the clinical expression of mutant mtDNA. PMID- 12768663 TI - [Examinations of distortion product otoacoustic emission in hereditary progressive non-syndromic hearing loss]. AB - OBJECTIVE: To evaluate the hearing function in patients with hereditary progressive non-syndromic hearing loss. METHODS: Distortion product otoacoustic emissions (DPOAE) and pure tone audiometry were carried out in 52 individuals from a family with non-syndromic hearing loss and 15 persons with normal hearing. RESULTS: 1. Sensorineural hearing loss (SNHL) was found in 34 individuals of the family. Among these individuals, DPOAE was totally absent in 15 cases (29 ears) with pure tone average > or = 40 dB and low amplitude or absent middle to high frequencies in 12 cases (23 ears) with high frequency hearing loss but pure tone average < or = 35 dB. 2. Among 21 individuals (42 ears) with normal audiograms, DPOAE presented lower amplitude or absent high and middle frequencies in 12 individuals. CONCLUSION: DPOAE can be used in identification of subclinical pathologic alterations in the cochlea. This would be of particular value in early diagnosis and genetic consultation. PMID- 12768664 TI - [The hereditary liability of otosclerosis and the strategies for treatment and prevention]. AB - OBJECTIVE: Otosclerosis is a disease of hereditary liability. It might be related to an autosomal dominant inheritance. The genetic penetrance is determined by multifactorial influences. This paper was designed to study the reasonable strategies for prevention and treatment of this disease. METHODS: A retrospective review derived from data of the management and follow-up of 14 cases in 6 families. According to the special pathological features of active phase of the otospongiosis, the suitable strategies for prevention and treatment of this disease were suggested. RESULTS: Diagnosis of 14 cases were confirmed by audiological, operative and/or pathological examinations. Eleven of fourteen cases underwent stapedectomy, in which 10 cases had the surgery on the ear with severer hearing loss, and one case on both sides. In all of the surgical cases, hearing levels improved significantly. Three cases have not yet received the stapedectomy, in which two cases scheduled for the surgery are currently treated by chondroitin sulfate and one case refused the surgical treatment due to financial shortage. CONCLUSION: Examination of the ear function at regular intervals (6-12 months) is recommended for the adults among the family members with high-incidence of otosclerosis. Audiological and radiological examination play an important role in early diagnosis and treatment. Attention should be paid to the medicine for management and prevention of this disease. Stapedectomy is an optimal choice for clinical otosclerosis not only due to its capacity to improve the hearing level but also to prevent the advance of hearing loss. Hearing aid is the suitable choice if stapedectomy is contraindicated. PMID- 12768665 TI - [Expression of heat shock protein 70 in immune response of the guinea pig inner ear]. AB - OBJECTIVE: To explore whether the immune response of the inner ear could induce heat shock protein (hsp) 70 in guinea pig cochlea. METHODS: A model of autoimmune inner ear disease (AIED) was established by systemically immunizing the guinea pig with the homologous crude inner ear antigen (CIEAg). The immunized cochleae and normal control cochleae were examined for the expression of hsp70 with techniques of immunohistochemistry and in situ hybridization. RESULTS: In the control animals, the expression of the hsp70-like protein appeared only in the spiral ganglion, whereas in the cochleae with CIEAg immunization, strong expression of the hsp70-like protein and its mRNA appeared in the spiral ganglion as well as in the stria vascularis and the spiral ligament. The hearing thresholds were significantly increased in 10 out of 28 cochleae (35.7%) with CIEAg immunization. CONCLUSION: The results suggest that the immune response of the inner ear can induce the expression of hsp70 in the guinea pig cochlea. PMID- 12768666 TI - [A novel method for screening anti-inner ear autoantibody in patients with autoimmune diseases]. AB - OBJECTIVE: To appraise the clinical value of a newly established method, rapid electric field immobilizing liquid phase molecule dot blot analysis (REILMD), for screening anti-inner ear autoantibody in patients with autoimmune diseases. METHODS: Seventy-one patients with 11 kinds of autoimmune diseases were chosen for the study. Both the general immunity and autoantibodies were tested. In the processes of detection of anti-inner ear autoantibody, REILMD was used for screening, and then the Western blot was used to define the molecular weight of inner ear antigen recognized by the positive autoantibody. RESULTS: Acceleration of erythrocyte sedimentation rate (ES), positive rheumatoid factor (RF), increases in C reactive protein (CRP), IgG and circulating immune complex (CIC) were found in most cases with rheumatoid arthritis (RA) and systemic lupus erythematodes (SLE). Some of these patients had increased IgA, IgM and C4. Two of 16 RA had anti-double-stranded DNA (dsDNA) and anti-mitochondria and 4/16 had anti-nucleus antibodies. Eleven of 16 SLE had antinucleus, 7/13 had anti ribonucleoprotein (RNP), anti-Sjogren syndrome A (SSA) and anti-dsDNA, 3/13 had anti-smooth muscle (Sm) and 1/13 had anti-DNA topoisomerase I (Scl)-70, striated muscle and stomach acid cell antibodies. No autoantibody was detected in AS. Anti inner ear autoantibody existed in 9 out of 71 patients (13%) with autoimmune diseases, in 2 of 21 patients (10%) with sudden deafness and only in 1 of 48 control subjects (2%, coronary heart disease). The anti-inner ear autoantibody was positive in 5 of 16 (31%) patients with SLE and 1 each in RA, AS, Behset's disease and streptococcus infection syndrome. In patients with positive anti inner ear antibody, 67% had anti-nucleus antibody, 50% had anti-RNP and dsDNA antibody. The molecules recognized by the positive anti-inner ear antibody were defined as 52,000, 36,000, 31,000 and 15,000 molecules of inner ear antigen. CONCLUSION: REILMD is a feasible and easy method for screening anti-inner ear autoantibody. Several autoimmune diseases, particularly SLE, may be implicated in damage to the inner ear. PMID- 12768667 TI - [Relationships between ankylosing spondylitis and ear disease]. AB - OBJECTIVE: To explore whether patients with ankylosing spondylitis (AS) have hearing loss and the pattern of hearing loss. METHODS: Questionnaire, physical examination of the ear, nose and throat, hearing test and sero-immunity examinations were conducted in 34 patients (68 ears) with AS. RESULTS: Among 34 patients, 11 cases reported decrease in the sound perception, and 24 cases (41 ears, 60.3%) had hearing loss as evidenced by the pure tone audiometry. In those patients with hearing loss, 2 ears with perforation of tympanic membrane showed conductive hearing loss and the remainder 39 ears showed sensorineural hearing loss in which 26 ears (66.7%) experienced high frequency hearing loss. Auditory brainstem responses were normally presented in 26 case, whereas abolished in 1 case (2 ears) with severe hearing loss. Examinations of sero-immunity showed a positive response in HLA-B27, but negative responses in the antinuclear antibody and rheumatoid factor. The percentage of the C-reactive protein increase was 81.8%. Autoantibodies anaginst the inner ear were positive in 9 cases (28.1%). In addition, all other immunological examinations revealed certain changes. CONCLUSION: More than half of the patients with AS had sensorineural hearing loss, particularly in the high frequency range. This hearing loss is paralleled by abnormal immunology and is a local expression of systemic autoimmune disease. Therefore, periodical hearing tests are necessary for these patients. PMID- 12768669 TI - [Clinical application of the multichannel auditory brainstem implant]. AB - OBJECTIVE: To investigate the surgical techniques and speech performance of multichannel auditory brainstem implant (ABI) in patients with bilateral acoustic neuromas (neurofibromatosis type 2). METHODS: The nucleus 21 channel auditory brainstem implant was implanted into the lateral recess of the fourth ventricle through the translabyrinthine approach in 7 patients after removal of the tumor. The accurate placement of electrode array was ensured by the electromyogram monitoring of the 7th and 9th nerves and the electrically evoked auditory brainstem responses (EABR). Initial switch-on occurred six weeks postoperatively. Speech evaluation was performed every 3 months for the first year and annually thereafter. RESULTS: During the surgery, the lateral recess could be found and the typical EABR could be recorded in 6 cases. They later reported a significant benefit from the device. Two of the cases have achieved functional open-set speech understanding. In contrast, one patient with no EABR because of difficulty of the anatomic location during the surgery had no sensations postoperatively. CONCLUSION: The multichannel ABI could effectively restore auditory sensations in patients deafened by bilateral acoustic neuromas. The accurate location of the cochlear nucleus complex during surgery was the key factor for the success of the operation. PMID- 12768668 TI - [The electron microscopic study of AchR and ultrastructure of neuromuscular junction in recurrent laryngeal nerve reinnervation]. AB - OBJECTIVE: The purpose of this study was to observe the distribution of Acetylcholine receptor (AchR) and ultrastructural changes of the neuromuscular junction after laryngeal reinnervation. METHODS: alpha-bungarotoxin-horseradish peroxidase (alpha-BT-HRP) conjugate electron microscopic immunocytochemical methods and computer analysis of intensity of immunocytochemical reaction were applied to observe the distribution and relative quantity of junctional N-AchR. RESULTS: The results showed that the synaptic nerve terminals and presynaptic membrane were segmentally disrupted and almost disappeared completely and were replaced by Schwann's cells at 3 weeks after neurorrhaphy. At 6 weeks, some of the regenerating axons grew to the neuromuscular junctions and all the muscle fibers were reinnervated at 12 weeks. But the original postsynaptic membranes were only partially in contact with the regenerated nerve terminals. These conditions persisted until 18 weeks after neurorrhaphy, when remyelination was found in the vicinity of the neuromuscular junctions. The N-AchR reaction product on the postsynaptic membrane were increased greatly about 1.28 times more than the normal at 3 weeks after neurorrhaphy. The reactive products decreased greatly which was only 0.47% of that in the normal at 6 weeks. It remained low until 12 weeks after neurorrhaphy, which was 0.72% of the normal side. It increased obviously to 0.83 times of normal sides at 18 weeks and didn't changed there after. CONCLUSION: The state of reinnervation determines the quantity of neuromuscular junctional N-AchR. PMID- 12768670 TI - [Long-term observation on two types of surgical operations for idiopathic hemifacial spasm]. AB - OBJECTIVE: To evaluate and compare the long-term curative effects of two types of surgical operations, microvascular decompression (MVD) and microneurovascular decompression neurocombing neurotraction draw (MVDCTD) for idiopathic hemifacial spasm (IHFS). METHODS: Five hundred and fifty-four patients with complete medical records and with at least 3-year followed-up had been collected since 1985. RESULTS: Of 148 MVD-treated cases with the longest follow-up of 13 years and 9 months, 117 cases were free of symptoms, giving a 79.05% cure rate. Thirty-one patients had recurrence of the symptoms, giving a 20.95% recurrence rate. Of 406 cases treated by MVDCTD with the longest follow-up of 12 years and 3 months, 374 were cured, giving a 92.12% cure rate, and 32 had recurrences of symptoms, giving a 7.88% recurrence rate. Most of them recurred within 2 years after the operation. Complications were sensorineural hearing loss in 31 patients (23 temporary, 8 permanent), temporary tinnitus in 22, temporary postoperative facial weakness in 78, and postoperative meningitis in 34 (3.13%, 33 cases were controlled with antibiotics and 1 patient died). CONCLUSION: The vascular compression at the root of the facial nerve is a main cause of IHFS, and the abnormal function of the facial nucleus is also one of the causes. MVDCTD for IHFS is characterized by its high curative rate, low recurrent rate and stable long-term effect, and is superior to the MVD. PMID- 12768671 TI - [Effects of intranasal budesonide on the infiltration and activation of eosinophils in nasal polyps]. AB - OBJECTIVE: To assess the effects of topical nasal steroid application on the infiltration and activation of eosinophils in nasal polyps. METHODS: Nasal polyps from intranasal budesonide treated patients (n = 16) and untreated patients (n = 16) were investigated. The samples were stained with histochemical and ABC immunohistochemical methods. RESULTS: The majority of eosinophils in nasal polyps were activated eosinophils. Compared with untreated polyps, the proportion of activated eosinophils (EG2+/eosinophil ratio) was significantly less in steroid treated polyps. Although the numbers of total eosinophils and actived eosinophils were lower in steroid-treated polyps, both of the differences were not statistically significant. CONCLUSIONS: It emphasized that the actived eosinophils play a key role in the pathogenesis of nasal polyps. The effects of intranasal steroid treatment on the number of eosinophils (total and activated) was more profound then on the proportion of activated eosinophils. PMID- 12768672 TI - [The lamina papyracea fracture reduction under nasal endoscope]. AB - OBJECTIVE: To evaluate the clinical results of blow-out laminapapyracea fracture reduction under nasal endoscopy. METHODS: All 6 cases had endophthalmos and blepharophimosis. Four cases also complained of diplopia. The horizontal and coronal CT of orbit showed the lamina papyracea fracture clearly. The degree and range of the lamina papyracea displacement was measured on CT films. The reduction under nasal endoscope was performed with local and topical anesthesia. We selected the Meeserklinger's route of entry. The uncinate process, ethmoidal bulla, and ethmoidal cells were removed successively. Then, with the help of a tonsil dissector, the lamina papyracea was pushed outward and replaced and was supported with iodoform gauze. After 2 to 3 weeks, the gauze was removed. RESULTS: The endophthalmos and diplopia disappeared in all cases. No postoperative complications occurred. With 0.5-2 years follow-up, the degree of the endophthalmos was stable within 2 mm. CONCLUSION: The reduction under nasal endoscope has many advantages such as short route of entry, simple performance, and no facial scar. It is a good operative option to correct the lamina papyracea fracture. PMID- 12768673 TI - [Horizontal-vertical hemilaryngectomy for advanced laryngeal cancer]. AB - OBJECTIVE: To evaluate the adequacy of horizontal-vertical partial laryngectomy in the treatment of advanced laryngeal cancer. METHODS: We retrospectively reviewed 57 patients with advanced laryngeal cancer (T3 or T4 on the primary site) treated by horizontal-vertical hemilaryngectomy. Fifty-six patients underwent neck dissections. RESULTS: The overall 3- and 5-year survival rates were 70% (40/57) and 64% (32/50), respectively. All patients had good phonation and resumed mouth-feeding after surgery. Among all the deaths, 4 died of recurrence of laryngeal cancer, 4 cervical metastasis, 4 distant metastasis, 2 intercurrent disease and 3 unknown. Of 36 patients with N0 neck, fifteen cases (15/36, 41.6%) had lymph node metastases pathologically. CONCLUSION: Horizontal vertical hemilaryngectomy is satisfactory for the treatment of selected advanced laryngeal cancers. It also preserves laryngeal functions in most cases. PMID- 12768674 TI - [Partial laryngectomy for T3 glottic carcinoma]. AB - OBJECTIVE: To explore the possibility of partial laryngectomy for T3 glottic carcinoma and selection of reconstruction methods. METHODS: From January 1986 to January 1994, 78 patients with T3 glottic carcinomas were treated by partial laryngectomy. The operative techniques and the reconstruction methods were retrospectively analyzed. RESULTS: The overall 3 and 5 years survival rates were 88.5% and 75.6% respectively. The local recurrence rate was 15.4%. The reconstruction methods for laryngeal defect repariring following partial laryngectomy were the use of pyriform sinus mucosa with thyroid perichondrium, epiglottic cartilage and bi-pedicaled and bi-muscular flap of strap muscle. The function of respiration, phonation and deglutition recovered well. CONCLUSION: The partial laryngectomy is suitable for T3 glottic carcinoma. The selection of reconstruction method is based on the size of laryngeal defect. PMID- 12768675 TI - [Descending cervical mediastinitis: report of 15 cases]. AB - OBJECTIVE: To analyze the cause, diagnosis and treatment of descending cervical mediastinitis. METHODS: Fifteen cases of descending cervical mediastinitis, which were treated from January 1985 to December 1997, were retrospectively reviewed. There were 10 males and 5 females, ranging in age from 2.5 to 82 years. RESULTS: The cause of descending cervical mediastinitis included odontogenic infection, suppurative tonsillitis, suppurative otitis media (cholesteatoma) complicating Mouret abscess, pharyngeal injury by foreign body and esophageal perforation. Different ways of drainage were adopted. Twelve cases were cured and three died. CONCLUSION: The key to successful management of descending cervical mediastinitis is early recognition, prompt and effective surgical drainage and appropriate antibiotics. PMID- 12768676 TI - [Laryngostroboscopy in the monitoring of precancerous lesion of vocal cord and in canceration tracing]. AB - OBJECTIVE: To evaluate the value of laryngostroboscopy in the monitoring of precancerous lesion of vocal cord and in the canceration tracing. METHODS: Fifty two cases of precancerous lesion of vocal cord were examined under Laryngostroboscope. The modifications of mucosal wave and amplitude of vocal cord vibration were observed. All cases were monitored for 1-6 years. RESULTS: The mucosal wave was weak in 45 of 52 cases (86.3%) and the amplitude of vocal cord vibration was weak in 4 of 52 cases (9.6%) with precancerous lesions. However, the mucosal wave disappeared and the amplitude of vocal cord vibration weakened in all 14 cases with early glottic cancer. CONCLUSION: Laryngostroboscopy can be used in diagnosis of glottic precancerous lesion and distinguishing it from the early glottic cancer. It is also an effective means of canceration tracing. PMID- 12768678 TI - [Upper airway occlusion sites of obstructive sleep apnea]. AB - OBJECTIVE: To locate the obstruction sites in patients with obstructive sleep apnea syndrome (OSAS) during sleep by MRI imaging and dynamic fiberoptic pharyngoscopy. METHODS: Fifteen OSAS patients confirmed by polysomnography (PSG) underwent ultrafast MR imaging during sleep. Their sequential midline sagittal and axial images were obtained and displayed in the cine mode. Then we examined the upper airway of the retropalate region (RP), retroglottal region (RG) and epiglottal region (EPG). We also did dynamic fiberoptic pharyngoscopy examination during sleep in 8 patients in another night. RESULTS: Among the 15 patients, 3 had only RP region obstruction, 2 had only RG region obstruction, and the other 10 had both RP & RG airway obstruction. In the 8 cases monitored with both MRI and fiberoptic pharyngoscopy, the results obtained from the 2 methods were identical. CONCLUSION: MRI and fiberoptic pharyngoscopy are useful in locating the pharyngeal airway obstruction sites of OSAS patients and would be helpful in selecting treatment measures. PMID- 12768677 TI - [Role of P53, nm23 proteins and vascular endothelial growth factor in angiogenesis and metastasis of laryngeal cancer]. AB - OBJECTIVE: To elucidate the role of P53, nm23 proteins and vascular endothelial growth factor (VEGF) on angiogenesis and metastasis of laryngeal cancer. METHODS: Specimens of 42 laryngeal carcinomas were studied by immunohistochemical staining for P53 protein, nm23 protein, VEGF and CD34 antibody. RESULTS: Positive expressions of P53 protein, nm23 protein and VEGF were found in 47.6%, 57.1% and 71.42% of laryngeal cancer specimens respectively. There was a positive relationship between P53 and VEGF expression. The microvessel density (MVD) in P53- or VEGF--positive tumors was significantly higher in patients with cervical lymph node metastases than those without metastases. No correlation was identified between nm23 and VEGF staining. The MVD in nm23-negative and VEGF- positive tumors was significantly high. These phenomena were more easily found in patients with cervical lymph node metastases than in patients without metastases. CONCLUSION: P53 gene play roles in the metastasis of laryngeal cancer through regulating VEGF expression and reacting MVD in tumor; and nm23 gene reacts through approaches other than regulating VEGF expression. PMID- 12768679 TI - [The role of heat shock protein 70 and its autoantibody in experimental autoimmune inner ear disease]. AB - OBJECTIVE: To examine the effect of antigen stimulation on induction of the heat shock protein 70(HSP70) in the guinea pig cochlea and to explore the role of HSP70 and its autoantibody in the pathogenesis and diagnosis of autoimmunize inner ear disease (AIED). METHODS: To establish the animal model of AIED, homologous crude inner ear antigen was used to immunize the guinea pig. The expression of HSP70 was detected by immunocytochemistry and the relative staining densities were quantified with a light microscope image analysis system. The autoantibodies against HSP71 in the plasma of animals were tested by Western blot using purified recombinant human HSP71 as antigen. RESULTS: HSP70 presented in the normal guinea pig cochlea at a lower level. After immunological challenge, the levels of HSP70 immunoreactivity in the immunized animals were significantly increased as compared with control animals. Optical densities of cochleae of immunized animals were significantly greater than those of animals in the control group (P < 0.01). Most of the immunized animals had developed autoantibodies against HSP71. The incidence of autoantibody in the experimental group was significantly different (P < 0.01) from the control group. CONCLUSION: Antigen stimulation could lead to an increase in the synthesis of HSP70 in the guinea pig cochlea. The detection of autoantibody against HSP71 might have significance in the diagnosis of AIED. PMID- 12768680 TI - [mRNA expression of hyaluronan synthase in the endolymphatic sac cells of guinea pigs]. AB - OBJECTIVE: To detect the mRNA expression of hyaluronan synthase in the endolymphatic sac cells of guinea pigs. METHODS: After consulting gene bank, we analyzed conservative sequence of hyaluronan synthases in different species, detected the mRNA expression of hyaluronan synthase in the endolymphatic sac cells of guinea pigs with oligonucleotide probe by hybridization in situ. RESULTS: mRNA of hyaluronan synthase was strongly expressed in some epithelial cells of endolymphatic sac, coupled with negative expression in negative control groups. CONCLUSION: It confirms that endolymphatic sac cells can synthesize hyaluronan. PMID- 12768681 TI - [Speech recognition and language development in prelingually deafened children with multichannel cochlear prosthesis]. AB - OBJECTIVE: To assess the development of hearing and language in prelingually deafened children with implanted multichannel cochlear prosthesis from the mandarin-speaking area, and to explore the basic pattern of speech recognition and language development as well as influencing factors. METHODS: Twenty-five prelingually deafened children and youths with cochlear implant participated in this study. Using "Evaluating method on hearing and speech rehabilitation in deafened children" as a test material. These materials consist of speech and environment sound discrimination, closed-set and open-set speech recognition and language development tests. RESULTS: All patients had the ability to hear sound and could discriminate different categorical sounds. Results indicated that the score of closed-set test exceeded the chance level. The correct recognition rate was escalating with prolonging the implanted time. Contrarily, the correct recognition rate was de-escalating as implanted age increased. Subjects having significant open-set speech recognition half year after the operation, indicated that the cochlear implant can promote prelingually deafened children to develop their speech level. CONCLUSION: Once the diagnosis of profoundly sensorineural hearing loss is made, the optimal treatment of choices is cochlear implant and speech rehabilitation. PMID- 12768682 TI - [Giant cell reparative granuloma of the skull]. AB - OBJECTIVE: To investigate the clinical course, histologic feature, diagnosis, differential diagnosis and treatment of giant cell reparative granuloma (GCRG). METHODS: Three cases of GCRG, which arose in the skull, were collected from 1987 to 1999 and analyzed clinicopathologically. RESULTS: Two patients had the lesion in the temporal bone and one in the orbit. One patient with GCRG had a recurrence after incomplete surgical resection. Histologically, all of three cases were characterized by fibroblastic proliferation with osteoclast-like giant cell rich areas, hemorrage, and newly formed bone or osteoid trabeculae. GCRG appeared as a typical bony lytic lesion on computed tomography (CT) and as low signal intensity on both T1 and T2 weighted images on magnetic resonance imaging (MRI). CONCLUSION: GCRG is an uncommon nonneoplastic lesion that typically arises in mandible and maxilla and its involvement in skull is rare. Although behaves benignly, GCRG may be locally aggresive. It is usually misdiagnosed for a giant cell tumor (GCT). Diagnosis of GCRG is based on typical histologic features as well as clinical information. Complete surgical resection is believed to be the most suitable treatment for GCRG and long-term follow-up by CT scanning is essential. PMID- 12768683 TI - [Nitric oxide and cytokines in otitis media with effusion]. AB - OBJECTIVE: To study the role of cytokines and nitric oxide (NO) in the pathogenesis of secretory otitis media (SOM). METHODS: The content of NO, interleukin-8 (IL-8) and tumor necrosis factor alpha (TNF-alpha) in blood plasma and middle ear effusion (MEE) were measured in 58 (80 ears) patients with secretory otitis media and in 23 normal subjects, respectively. RESULTS: NO, IL-8 and TNF-alpha were found in 100%, 82.2% and 71.4% of MEE respectively. NO, IL-8 or TNF-alpha of blood plasma in the SOM group was higher than those in the normal control group (P < 0.05). The concentration of NO, IL-8 or TNF-alpha in MEE was remarkably higher than those in blood plasma (P < 0.01). It revealed that the shorter the course, the higher the concentration of IL-8 in MEE (P < 0.01), and the longer the course, the higher the concentration of NO or TNF-alpha in MEE (P < 0.05). IL-8 concentration of MEE in serous fluid group was remarkably higher than that in the mucous fluid group (P < 0.01), and the concentration of NO or TNF-alpha of MEE in the mucous fluid group was higher than that in the serous fluid group (P < 0.05). NO level of MEE was positively correlated with TNF-alpha (r = 0.696 P < 0.01). CONCLUSION: NO IL-8 and TNF-alpha might be important mediators in MEE secretion in SOM. IL-8 might participate in the defensive reaction of organism during the early stages of SOM, and play a role in serous effusion secretion. NO and TNF-alpha might be closely related to the persistence of SOM, and might be a mediator of mucin secretion in SOM. PMID- 12768684 TI - [Application of extended high frequency audiometry in the early diagnosis of noise--induced hearing loss]. AB - OBJECTIVE: To assess the application of 10-20 kHz audiometry in early diagnosis of noise-induced hearing loss and to explore the relationship between hearing loss and subjects' age, the exposure time as well as the noise level. METHODS: One thousand workers with noise exposure history were examined by pure tone audiometry(0.5-20 kHz). One hundred and twenty normal subjects served as the control group. RESULTS: The noise levels at working sites ranged from 95 to 115dB(A). Compared with the control group, noise-exposed subjects showed minor changes in the thresholds from 0.5 to 6 KHz, whereas the thresholds were elevated in the high frequency range of 10-16 kHz (P < 0.001). The incidence of ears without any response to the maximum output at the frequency range of 14 to 20 kHz was much higher (P < 0.01) than that of the control group. In the animals exposed to noise, the thresholds of 10-18 kHz increased with the aging and with increase of working time (6-10 years), but there were no significant differences among subjects exposed to the noises at different levels(P > 0.05). CONCLUSION: In noise-exposed ears, the occurrence of threshold changes in the high frequency range (10 to 20 kHz) is earlier than that in the low frequency range (0.5 to 6 kHz). Aging and working time also affect the 10-20 kHz hearing thresholds. The lack of responses to maximum output and the changes in thresholds in the high frequency range (10-20 kHz) can be used as indices for the diagnosis of early noise-induced hearing loss and for the assessment of the susceptibility of the subject to noise damage. PMID- 12768685 TI - [The outcome of 1268 cases of endoscopic sinus surgery]. AB - OBJECTIVE: To evaluate the curative effect of endoscopic sinus surgery (ESS). METHODS: One thousand two hundred and sixty-eight cases with chronic sinusitis and nasal polyps undergone ESS from 1989 to 1997 were appraised with the standard criteria (Chinese ENT Association, 1997). There were 366 cases(28.9%) in type 1, 646 cases(50.9%) in type 2 and 256 cases(20.2%) in type 3. One thousand two hundred and sixty-two cases underwent ESS under intranasal local anesthesia and sedation; 182 cases underwent submucous resection of nasal septum or septoplasty. Long-term follow-up was performed at 9 to 42 months(mean = 15 months). RESULTS: The results showed that cure rate was 93.7% for type 1, 81.6% for type 2 and 68.4% for type 3. There was significant difference in cure rate among different types (P < 0.01). Complication of ESS in 1,268 cases occurred in 2.3%. CONCLUSION: ESS is efficacy and safe in the treatment of nasal polyps and chronic sinusitis. The cure result is directly related to the types of the disease. PMID- 12768687 TI - [Pedicled galeal flap in the reconstruction of head and neck tumor defects]. AB - OBJECTIVE: To evaluate the advantage and applications of pedicled galeal flap in head and neck region. METHODS: A consecutive series of 17 patients underwent surgical reconstruction with pedicled galeal flaps after head and neck tumor resection. The defects included nasopharynx, skull base, maxilla, orbital base, oropharynx and oral cavity and the size ranged from 5 cm x 5 cm to 10 cm x 10 cm. The technique for using this flap was described and application was illustrated with 3 case reports. RESULTS: Complete success of galeal flaps for the reconstruction of head and neck defects was achieved in 13 of the 17 cases (76.5%) and partial necrosis was observed in the remaining 4 cases (23.4%). Immediate wound complications occurred in four cases, which resolved spontaneously. Four delayed complications were observed in 4 of 9 survival cases that included trismus (3) and alopecia (1). CONCLUSION: Galeal flap is a thin, pliable and well vascularised reconstruction tissue and is highly reliable. The donor site morbidity is minor. We have found the flap to be useful in the reconstruction of a variety of defects in head and neck, especially in skull base, orbital base, nasopharynx and oropharynx. PMID- 12768686 TI - [Partial laryngectomy plus radiotherapy versus partial laryngectomy alone for laryngeal carcinoma]. AB - OBJECTIVE: To investigate the effect of radiotherapy in the combined treatment of laryngeal cancer with partial laryngectomy. METHODS: One hundred and seventy-one cases of laryngeal cancer were randomly separated into three groups of partial laryngectomy, partial laryngectomy with preoperative radiotherapy (dose 40-50Gy) and partial laryngectomy with postoperative radiotherapy(dose 51-70 Gy). The surgical procedures included cordectomy, vertical partial laryngectomy, horizontal partial laryngectomy and near total laryngectomy. RESULTS: The overall 5-year survival rate was 82% (123/150). The 5-year survival rates for surgery alone group (56 cases), surgery with preoperative radiotherapy group(65 cases) and surgery with postoperative radiotherapy group(50 cases) were 85.7%, 80.7% and 79.5% respectively. There was no significant difference among the groups(chi 2 = 0.703, P = 0.704). Analysis of survival rates revealed a significant difference among the different stages(chi 2 = 12.248, P = 0.007). All 171 cases except 3 that had near total laryngectomy achieved satisfactory phonation. CONCLUSION: Neither preoperative radiotherapy nor postoperative radiotherapy can improve the 5-year survival rate of laryngeal cancer with partial laryngectomy. All patients with partial laryngectomy got satisfactory phonation. PMID- 12768688 TI - [Supraglottic horizontal partial laryngectomy]. AB - OBJECTIVE: To evaluate the long-term result of supraglottic horizontal partial laryngectomy. METHODS: Fifty-five patients with supraglottic laryngeal carcinoma were treated surgically by supraglottic horizontal partial laryngectomy from 1986 to 1995. There were 32 males and 23 females with an average age of 56. Eleven cases were staged I, 26 staged II, 11 staged III and 7 staged IV. Closure of the surgical defect begins by covering the thyroid cartilage with mucosa of the medial wall of the pyriform sinus. The outer perichondrium of thyroid cartilage was approximated with the mucosa of laryngeal ventricle. The hyoid cartilage was excised. The base of tongue was sutured to the reserved thyroid cartilage to take the place of epiglottic cartilage. RESULTS: The 3- and 5-year survival rates were 85% (47/55) and 77% (37/48) respectively. The decannulation rate was 96%. Most cases had acquired good speech and swallow function after 8-30 days. Complications included granulation and polyp formation at the base of tongue or at the anterior commissure in 4 cases. CONCLUSIONS: Supraglottic horizontal partial laryngectomy is very effective both in eradicating disease and in preserving laryngeal function. PMID- 12768689 TI - [The application of epiglottis in laryngeal function reconstruction after partial laryngectomy]. AB - OBJECTIVE: To observe the appliance value of epiglottis in laryngeal function reconstruction after partial laryngectomy. METHODS: Thirty-two cases of glottic and superglottic carcinomas of larynx had conducted partial laryngectomy and the laryngeal function was reconstructed using epiglottis from December. 1992 to January 1996. RESULTS: Nineteen of the twenty-one glottic carcinomas and five of eleven supraglottic carcinomas were decannulated and recovered with laryngeal functions. All the patients rehabilitated deglutition in 14-18 days after surgery. None presented complication of aspiration. CONCLUSIONS: Epiglottis has many advantages in partial laryngectomy and the laryngeal reconstruction. It is easy to approach, apply and heal in short period. It also prevents aspiration and improves the decannulation rate as well as the quality of life in partial laryngectomy patients. PMID- 12768690 TI - [The clinical application of stereography in laryngology with "the three dimensional reconstruction system of medical faultage photograph"]. AB - OBJECTIVE: To study the value of the clinical application of stereography in laryngology. METHODS: Taken by Microtek E6 scanner, the CT faultage images were inputted in personal computer and reconstructed to a three-dimensional stereograph by "the three-dimensional reconstruction system of medical faultage photograph". The software was compiled using IDL5.2, a complete, integrated software environment for data analysis, visualization and application. RESULTS: The three-dimensional laryngeal graph could be incised in random, displayed in optional degree, and was rotatable, semitransparent and measurable. The tumor could identified be identified by a special color and revealed through translucent parenchyma and cartilage. Reconstruction stereographs of laryngeal and hypopharyngeal cancer of 26 patients were investigated. CONCLUSION: It is convenient for popularization because the three-reconstruction process is built on a personal computer without the need of any equipment for image acceleration. It is very helpful for ENT surgeon to define the range of the tumor, to fix on surgical pattern accurately and to anticipate the operational course. PMID- 12768691 TI - [Use of Hopkins rod-lens coupled with grasping forceps sheaths for extraction of tracheobronchial foreign bodies]. AB - OBJECTIVE: To study the clinical application of Hopkins rod-lens coupled with gasping forceps sheaths in the extraction of tracheobronchial foreign bodies. METHODS: Sixty-two patients with tracheobronchial foreign bodies were operated on with Hopkins rod-lens coupled with grasping forceps sheaths under direct telescopic view or TV watching. High frequency jet ventilation was used and BPs ECGs SaO2 were monitored in all cases. RESULTS: Sixty-two patients were all successfully operated on. The extraction was primarily successful in 90% of the cases; and secondarily successful in 7% of the cases. No death and serious complication occurred. CONCLUSION: Compared with conventional bronchoscopic foreign body extraction, this new instrument and technique make it possible for the operator to clamp and extract foreign body under direct telescopic view or TV watching. It has increased the accuracy and safety of bronchoscopic extraction of tracheobronchial foreign bodies. PMID- 12768692 TI - [The upper airway MRI of obstructive sleep apnea patients]. AB - OBJECTIVE: To investigate the mechanisms of obstructive sleep apnea syndrome(OSAS) through observing the upper airway caliber and its corresponding pharyngeal wall of OSAS patients by MRI. METHODS: The upper airway(UA) of 18 OSAS patients were examined by MRI. The cross-sectional area of the UA and lateral parapharyngeal fat pad (LPFP) were calculated. The thickness of lateral and posterior pharyngeal wall (LW & PW) and other related indices were also measured. Nineteen nonsnoring age-matched normal subjects were selected as the control group. RESULTS: 1. The retropalatal (RP) and retroglossal (RG) region UA cross sectional areas of OSAS patients were smaller than that of the control (P < 0.05). The epiglottal (EPC) region UA cross-sectional area was not significantly different between the 2 groups (P = 0.204). 2. The anterior-posterior/Lateral ratio(A-P/L) of UA in RP, RG, EPG regions was bigger in patients group than that of the control (P < 0.05). 3. The cross-sectional area of LPFP in patients group was larger than that of the control group (P = 1.76E-7). 4. The thickness of the PW in the RP, RG and EPG region was thicker in patients group than that of the control (P < 0.05). Although the thickness of the LW in the RP region was not statistically different between the 2 groups (P = 0.94), this index in RG and EPG region was larger in patients group than that of the control (P = < 0.05). 5. Sagittal MRI image showed that the length, thickness and cross-sectional area of the palate in patient group was larger than that of the control (P < 0.01). CONCLUSIONS: The study suggests that the pathogenesis of OSAS is related to the following factors: UA caliber and its AP/L ratio, the thickness of LW and PW, the size of LPFP in RP region, and the size and length of the palate. PMID- 12768693 TI - [The position of submaxillary transcutaneous electrical stimulation for obstructive sleep apnea syndrome]. AB - OBJECTIVE: To investigate the effective position of submaxillary transcutaneous electrical stimulation for obstructive sleep apnea syndrome (OSAS). METHODS: With fixed provocative locus of genioglossus and its controlling nerve, 11 normal healthy men and 9 patients with OSAS during awakening were studied to disclose the effect of submaxillary constant-current stimulation under different stimulative radius or intensity and thickness of submaxillary skin. The effects of electric stimulation in 9 OSAS patients with narrowing of the retrotongue-base pharynx level were compared with the control during sleep. RESULTS: The submental and internal side of mandibular angle was effective loci for stimulating genioglossus. Stimulative radius and intensity of both loci were not significantly different (P < 0.05) and no influence of the skin thickness was found. During sleep, indices of breath disorder (P < 0.05), oxygen saturation (P < 0.05) and clinical symptoms had significantly improved. CONCLUSION: Submaxillary electrical stimulation with fixed provocative locus is effective for the treatment of OSAS. The mechanism is perhaps stimulation of genioglossus pushes the tongue ahead and opens the pharyngeal cavity. PMID- 12768694 TI - [Acoustic analysis and characteristics of vocal range in Beijing Opera actors]. AB - OBJECTIVE: To get the objective acoustic parameters of the voice of Beijing Opera actors and set a foundation for the training and protection of the special professional voice. METHODS: Seventy-three (age 16-57 years) professional actors and students were asked to produce sustained comfortable vowels /a/ and /i/, and to sing two pieces of songs which were in the category of Xipi and Erhuang respectively. Dr. Speech for windows version 3.0 was used to get the acoustic parameters of the vowels and the songs. RESULTS: F0 of the vowels /a/ and /i/ of different Hangdangs were Chou (272.6 +/- 42.0) Hz (mean +/- s), (304.2 +/- 22.1) Hz; Xiaosheng (499.3 +/- 34.0) Hz, (485.4 +/- 18.7) Hz; Laosheng (335.6 +/- 60.0) Hz, (317.9 +/- 45.1) Hz; Hualian (319.0 +/- 61.3) Hz, (340.1 +/- 68.8) Hz; Laodan (427.6 +/- 47.2) Hz, (437.7 +/- 45.8) Hz; Huadan (535.8 +/- 48.8) Hz, (561.6 +/- 29.2) Hz; Qingyi (548.0 +/- 69.5) Hz, (543.5 +/- 79.3) Hz; these and other acoustic parameters of vowels such as Jitter, Shimmer and NNE were all within the normal range given by the software. The vocal range of Beijing Opera actors was from 1.7 to 2.8 oct, and most of the highest and the lowest pitches were higher than that of tenor or soprano. CONCLUSIONS: These findings may help to provide insight regarding the acoustic characteristics of the voice of Beijing Opera actors. PMID- 12768695 TI - [An epidemiological investigation of anotia and microtia in China during 1988 1992]. AB - OBJECTIVE: To investigate the epidemiological characteristics of cases with anotia and microtia in China. METHOD: The birth defect monitoring program was undertaken by a hospital-based monitoring method in 443-588 hospitals from 30 provinces, cities and autonomous regions across China. Data including intrauterine death and stillbirth from 28 weeks of gestation to a period of 7 days postpartum were collected between 1988 and 1992. RESULTS: A total of 3,246,408 births was monitored from 1988 to 1992, in which 453 cases had anotia and microtia. The average incidence was 1.40 per 10,000 during the five-year period. The decreased tendency of incidence was noted during the period (chi 2 = 5.5588, P < 0.05). The incidence in the urban area was significantly higher than that in the rural area. There was no sex difference in the incidence of anotia and microtia. The incidence varied among 30 provinces with highest incidence in Xinjiang province (2.08 per 10,000 births) and lowest in the Inner Mongolia Autonomous Region (0.33 per 10,000 births). In cases with the defects of anotia and microtia, 60.4% were accompanied with other congenital malformations. The proportion of multiple malformations with microtia and anotia was significantly higher than that of isolated case (chi 2 = 36.9277, P < 0.01). The highest incidence of concurrent malformations was anophthalmia or microphthalmia (13.1%), followed by facial cleft (12.6%), neural tube defects (10.0%), limb reduction defects (9.6%) and polydactyly (5.4%). There were also 3.5% cardiac defects. CONCLUSIONS: The prevalence of anotia and microtia varied among provinces across China. The high proportions of cases with anotia and microtia had multiple malformations. Therefore, careful examination of other malformations in patients with anotia and microtia is necessary. PMID- 12768696 TI - [Expression of substance P receptor mRNA in nasal mucosa of rat in allergic rhinitis model]. AB - OBJECTIVE: To study the expression of substance P receptor messenger RNA (SP-R mRNA) in nasal mucosa in allergic rhinitis (AR) rat model. METHODS: Twenty Wistar rats free of disease were randomly divided into two groups. AR model rats were established through repeated intraperitoneal shot of ovalbumin (OV) and consequently confirmed by local challenge with OV. SP-R mRNA in nasal mucosa, obtained from two groups, were used to do reverse transcriptive polymerase chain reaction (RT-PCR) and Southern blot. beta-actin was used as a standard control through out the whole process. RESULTS: The results showed definitely that there were positive expression of SP-R mRNA in normal nasal mucosa. This expressions increased significantly (P < 0.05) during nasal allergy. CONCLUSION: Increased expression SP-R mRNA in nasal mucosa in AR model might play roles in the development of AR. PMID- 12768697 TI - [The concentration and expression of IL-5 in human nasal polyp tissue]. AB - OBJECTIVE: To study the concentration and expression of IL-5 in nasal polyp tissues and explore its significance in the micro-environment differentiation of eosinophils accumulation and clarify the conception of nasal polyposis. METHODS: The concentration and expression of IL-5 in nasal polyp tissues of 40 patients were determined by ELISA and immunohistochemistry, and inferior turbinate mucosa from patients with nasal polyps and healthy volunteers was used as control. RESULTS: 1. IL-5 concentration in the polyp tissues was significantly higher than that in inferion turbinate mucosa(P < 0.05). There was no significant difference in inferion turbinate mucosa between the patients with nasal polyps and healthy volunteers (P > 0.05). IL-5 concentration in polyp tissues was markedly higher in patients with extensive polypoid change of nasal mucosa, history of previous polypectomy and allergic rhinitis compared with those without these features (P < 0.05). IL-5 concentration had no correlation with age and sex (P > 0.05). 2. 80.1% of the eosinophils were positive for IL-5 and 90.9% of IL-5 positive cells were eosinophils. Only 3.7% of the lymphocytes and neutrophils were IL-5 positive, and IL-5 was not detectable in epithelial cells. IL-5 expression in eosinophils of polyp tissues was remarkably stronger than that of the turbinate mucosa (P < 0.05). There was no significant difference in inferion turbinate mucosa between the patients with nasal polyps and healthy volunteers (P > 0.05). IL-5 expression of eosinophils in polyp tissues was significantly stronger in patients with extensive polypoid change of nasal mucosa, history of previous polypectomy and allergic rhinitis compared with those without these features (P < 0.05). There was no significant difference in IL-5 expression in lymphocytes and neutrophils between polyp tissues and inferior turbinate nasal mucosa (both P > 0.05). CONCLUSION: IL-5 is a key protein in eosinophilic pathologic mechanisms in nasal polyp tissues. PMID- 12768698 TI - [Significance and detection of interleukin-8 in human nasal polyps]. AB - OBJECTIVE: To investigate the distribution and content of immunoreaction interleukin-8(IL-8) antigen in the human nasal polyps. METHODS: Twenty-two cases of nasal polyp and 10 cases of normal inferior turbinate were studied with immunohistochemical SABC method and ELISA method. RESULTS: Immunohistochemical results demonstrated that IL-8 antigen staining occurred predominantly within inflammatory and epithelium cell plasm. The level of IL-8 in nasal polyp tissue was higher than in normal control specimens. The value in nasal polyp tissue was in 322.06-2091.41 pg/ml, but in normal with a range of 29.31-332.19 pg/ml. The results demonstrated a remarkable difference between the two groups (P < 0.001). CONCLUSION: These data demonstrated the presence of IL-8 antigen in nasal polyps. Thus IL-8 likely plays a significant role in the pathogenesis of this disease process. There is an important significance of finding a potential treatment method. PMID- 12768699 TI - [The secretion of chemokine RANTES in epithelial cells of nasal polyps and its significance]. AB - OBJECTIVE: To investigate the effect of RANTES(regulated upon activation, normal T cell expressed and secreted) on chemotaxis, migration and accumulation of eosinophils in nasal polyps during epithelial immune responses. METHODS: The epithelial cells were obtained from nasal polyps and inferior turbinates in patients with snoring, and cultured for 24 and 48 hours with IL-1 beta(25 micrograms/L, 50 micrograms/L). RANTES was measured in the culture supernatant by ELISA. RESULTS: 1. On incubation with IL-1 beta, the epithelial cells from inferior turbinate and nasal polyps released 1.5-10 fold and 10-20 fold greater amounts of RANTES respectively than unstimulated samples. Significant increase in levels of RANTES was found in both groups(P < 0.001); 2. There was no difference between nasal polyp and inferior turbinate before incubation with IL-1 beta(P > 0.05) in the level of RANTES. After IL-1 beta stimulation, the concentration of RANTES was higher in polyp than in inferior turbinate (P < 0.001); 3. The expression of RANTES in both groups was dose-dependent and time-dependent upon stimulation of IL-1 beta. CONCLUSION: The epithelial cells of nasal mucosa are active end-organs, capable of releasing chemokine RANTES. In nasal polyps, RANTES has chemotactic activity for eosinophils and other inflammatory cells. It could affect the chemotaxis and eosinophil function and may play an important role in polyp formation. PMID- 12768700 TI - [Diagnosis and nasal endoscopic surgery of rhinogenic optic function damage]. AB - OBJECTIVE: To explore the diagnosis and the surgical treatment of rhinogenic optic function damage through transnasal endoscopic approach. METHODS: The clinical characteristics and the results of transnasal endoscopic surgery of 32 cases(32 eyes) with rhinogenic optic function damage were analyzed. RESULTS: Eleven eyes were cured, 20 eyes were improved, 1 eye failed, No recurrence was found within 6 months to 3 years after operation. CONCLUSION: The sinus inflammatory disease might injure optic function. Early diagnosis as is important and the endoscopic sinus surgery combined with appropriate medical drug administration are in effective. PMID- 12768701 TI - [Sodium salicylate induced intracellular Ca2+ concentration change in guinea pig outer hair cells]. AB - OBJECTIVE: In order to explore the mechanism of sodium salicylate ototoxicity the effect of sodium salicylate on intracellular Ca2+ ([Ca2+]i) of the outer hair cells (OHCs) and the effects of calcium channel antagonist on sodium salicylate induced [Ca2+]i changes of the OHCs were examined. METHODS: The OHCs of guinea pig cochlear were isolated using an enzyme-machine methods and loaded with 10 mumol/L Fluo-3/AM for 30 min at 37 degrees C. Individual Fluo-3 loaded OHC was examined with a confocal microscope (ACAS Ultima, USA) using a 20 x objective lens and linear scan mean. The fluorescent images, collected each 5-sec for 300 sec, were stored in a computer. The fluorescent intensity of the OHCs were analyzed by the software cooperated with the confocal microscope, and a curve of fluorescent intensity changes trend against time was obtained. RESULTS: The [Ca2+]i of OHCs were steady under normal extracellular liquid perfusion. Salicylate increased [Ca2+]i in OHCs in calcium-free medium(10/10) and standard medium (8/10). 3 mmol/L lidocaine inhibited [Ca2+]i increase in OHCs induced by salicylate(0/10). The [Ca2+]i increase in OHCs induced by salicylate could be blocked by flunarizine(3/12). Nimodipin failed to block the [Ca2+]i increase of OHCs induced by salicylate (7/7). CONCLUSION: Sodium salicylate can result in [Ca2+]i increase in OHC markedly, which may be due to the Ca2+ release from intracellular calcium stores. The effects of salicylate on [Ca2+]i in OHCs might be one of the mechanisms of salicylate ototoxicity. PMID- 12768702 TI - [Amelioration of nerve growth factor against noise-induced threshold shift: a transmission electron microscope observation]. AB - OBJECTIVE: To study the protective effects of nerve growth factor(NGF) on noise induced hearing damage in guinea pigs. METHODS: NGF injected Guinea pigs were consecutively exposed to white noise of 115 dB(A) for 6 days continually (45 min.d-1). Auditory thresholds were measured using auditory cortex evoked response to tone bursts in different post-exposure intervals (1 h, 1 d, 2 d, 3 d and 6 d). The ultrastructural changes within hair cells were also observed by a transmission electron microscope(TEM). RESULTS: The auditory threshold shifts in test group A(NGF:1,000 U.kg-1.d-1, i.m.), B(NGF:2,000 U.kg-1.d-1, i.m.) and C(NGF:3,000 U.kg-1.d-1, i.m.) were significantly fewer than that in the control group(Saline: 1 ml.kg-1.d-1, i.m.). Threshold shifts almost recovered in test group B and C 3 days after the exposure; while a threshold shift of (16.43 +/- 6.91) dB was present 6 days after the exposure in the control group. TEM showed that all three rows of the outer hair cells(OHCs) of the basal turn in the control group displayed significant pathological changes. Depolymerization of actin filiaments within stereocilia, swelling of submembraneous cistern and the efferent nerve-ending and slight edema of hair cells were evident. In test group A, the hair cells display slight pathological changes, which are confined in the third row of OHCs in a local position of the basal turn. In group B and C hair cells have nearly normal appearance. CONCLUSION: NGF is able to reduce threshold shift, and promote the recovery of auditory threshold in acoustic trauma. This factor can, to some extent, protect against noise-induced hearing damage. PMID- 12768704 TI - [CT anatomic measurement of the optic canal and its clinical significance]. AB - OBJECTIVE: To probe into the normal anatomy of the optic canal and its clinical significance. METHODS: The optic canals of 200 normal subjects were examined by thin layer CT scan in axial nd coronal positions. The distance and anatomic relationship with circumferentral structure were measured. RESULTS: In average: (1) The transverse diameter of the optic canal was (3.57 +/- 0.61) mm, the longitudinal diameter was (4.82 +/- 0.38) mm, the length of the inner wall was (12.62 +/- 2.59) mm, the length of the outer wall was (10.18 +/- 2.19) mm, and the length of the optic canal was (11.46 +/- 2.35) mm. The angle between optic canal and the middle line of the skull was 34.34 degrees +/- 6.48 degrees, the angle between optic canal and the intraorbital optic was 10.69 degrees +/- 6.48 degrees. The thickness of the inner wall was (0.68 +/- 0.46) mm. It showed no significant difference between the two sides of the optic canal in diameters(P > 0.05). (2) The optic canal, the superior orbital fissure and the inferior orbital fissure were respectively shown as track, crack and gorge type in CT axial pictures, while in the coronal pictures, they were respectively shown as butterfly eyes, horizontal fissure and vertical fissure type. (3) The relationship of the posterior ethmoid sinus and the optic canal was: anterocanal(42.00%), semicanal(27.5%), whole-canal(18.75%), sella turcica(9.75%) and circumcanal(2.00%). (4) The relationship of the position of the sphenoidal sinus and the optic canal was: I degree sphenoidal sinus(20%), II degree (63%) and III degree (17%). (5) The incidence of the bony defect of the inner optic canal wall was 25.5%. The range of the defect was (5.07 +/- 2.52) mm. CONCLUSION: It's a reliable way to show the relation of the optic canal and circumcanal by CT, and it is of important value in operation. PMID- 12768703 TI - [Otogenic hypertrophic cranial pachymeningitis associated with edema of the temporal lobe and organic mental disorder-case report]. AB - OBJECTIVE: Hypertrophic cranial pachymeningitis(HCP) is a rare disease which might be misdiagnosed. One typical case of otogenic hypertrophic cranial pachymeningitis (HCP) associated with edema of the temporal lobe and organic mental disorder was reported. Literatures associated with HCP were reviewed for reference. METHODS: In July, 1997, a 26-year-old man was admitted for right-sided severe headache, dizziness, nausea, vomiting, facial and abducens nerve palsy. MRI and CT revealed inflammation in the temporal bone and locally thickened dura mater with obvious enhancement as a band in the petrous apex and nearby tentorium cerebellum. The patient then underwent operation including decompression of the facial nerve, labyrinthectomy, and eradication of the inflammatory cells in the mastoid and petrous portion. Postoperatively, the clinical signs disappeared except hearing loss. Seven months later, the patient suffered from mental disorder with interrupted excitement to hit and abuse the family members. He was then treated in a psychosis hospital but no improvement could be seen. On May 13, 1998, MRI and the enhanced MRI demonstrated diffuse thickened tentorium cerebellum and dura mater in the middle cranial fossa. The inferior portion of the temporal lobe was obvious edema. The internal carotid artery was partially occluded due to the thickened wall of cavernous sinus. RESULTS: Hence, the diagnosis of HCP associated with edema of temporal lobe and organic mental disorder was established. The patient was cured by high dosage of penicillin. CONCLUSION: MRI is an essential method for diagnosis of HCP. Antibiotic was an effective treatment. PMID- 12768705 TI - [The study about the characteristics of upper airway in obstructive sleep apnea syndrome]. AB - OBJECTIVE: To investigate the upper airway (UA) characteristics of the patient's with obstructive sleep apnea syndrome (OSAS) and analyses the value of computer assisting fiberoptic pharyngoscopy with muller's maneuver (CFPMM) in topodiagnosis of OSAS. METHODS: The pharyngxes of 30 cases with OSAS and 30 controls were examined by CFPMM. The cross-section area at velo- and tongue pharyngeal, pharyngeal collapsibility, sites of UA obstructive and their anatomical factors were measured and calculated and the correlation between the anatomical abnormalities of UA and outcomes of polysomnography (PSG) were analysed. RESULTS: 1. CFPMM can accurately measure and calculate the cross section area of the upper airway. 2. mean areas of velopharyngeal and tongue pharyngeal were significantly less than those in control subjects (all P < 0.05). OSAS patient's also showed relatively much larger collapsibility at sach site of the UA than controls. 3. At muller's maneuver, all the areas of OSAS patient's were below 40 mm2 and 60% cases appeared complete occlusion in pharyngeal. 4. All patient's had obstruction at velopharymx and airway collapses at multiple sites in 40% of cases. 5. At velopharyngeal the anatomical factors causing obstruction mainly were posterior displacement of soft palate, thicken and collapse of the pharyngeal wall. Besides the redundant lymph tissue at tongue base and posterior displacement of the tongue base, enlarged inferior pole of tonsil and collapse of pharyngeal wall played an important role at tongue-pharyngeal obstruction. 6. Diminished pharyngeal apertures and collapsibility were associated with increased rates of apnea and hypopnea index (P < 0.05). CONCLUSION: 1. CFPMM is able to measure and calculate UA's cross-section area and collapsibility, determine the site of obstruction, and helping the treatment. 2. Complete occlusion of UA at muller's maneuver is the criterion to determine site of UA obstruction in OSAS patient's during waking hours. 3. The anatomical abnormalities of UA of patients is reflects the severity of the OSAS. PMID- 12768706 TI - [Laryngeal function-sparing surgery for pyriform sinus cancer]. AB - OBJECTIVE: To evaluate the oncologic effect of laryngeal function-sparing surgery for the patients with pyriform sinus carcinoma. METHODS: A retrospective analysis was performed. Forty-four patients with pyriform sinus cancer who underwent operations were included. There were 10 patients in T1, 14 in T2, 18 in T3 and 2 in T4 categories. 42 patients received preoperative radiation and 2 cases, post operative radiation. Surgery for the local lesions: 36 patients underwent pyriform sinusectomy and 8, partial laryngectomy and pyriform sinusectomy. RESULTS: Five year survival rate is 50% for the whole series, 80% for stage I, 71.4% for stage II, 52.9% for stage III, 26.7% for stage IV and 45% for all T3 + T4 patients. Local control rate is 81.8% for all patients and 75% for T3, T4 patients. CONCLUSION: For selected pyriform sinus cancer patient a conservative laryngeal surgery is warranted with the combination of preoperative radiative therapy. PMID- 12768707 TI - [The study of human papilloma viruses infection in juvenile pharynlaryngeal papillomatosis by PCR and dot blot hybridization]. AB - OBJECTIVE: To study the relationship between infection of human papillomavirus and juvenile pharynlaryngeal papilloma. METHODS: Polymerase chain reaction (PCR) and dot blot hybridization were used to detect HPV6,11,16,18,33 DNA of 35 samples of pharynlaryngeal papilloma and 10 samples of vocal nodule. RESULTS: The positive rate of HPV in pharynlaryngeal papilloma was 91.4% (30/35). Among them the positive rates of HPV6 and HPV11 were 54.2%(19/35) and 25.7%(9/35), the positive rate of multiple types of HPV6 + 11 was 11.4%(4/35). The positive rate of HPV16,18,33 was negative. The positive rate of HPV in vocal nodule was negative and were significantly different from that of the pharynlaryngeal papilloma group. CONCLUSION: The results suggest that infection with HPV, especially with HPV6, is closely associated with the development of juvenile pharynlaryngeal papilloma in Wenzhou area. HPV-DNA detected by PCR and dot blot hybridization has highly specificity and sensitivity. PMID- 12768708 TI - [The study on the role of telomerase activity in human nasopharyngeal carcinoma]. AB - OBJECTIVE: To elucidate the role of telomerase activity in nasopharyngeal carcinoma (NPC). METHODS: Telomerase activity was detected using the telomere repeat amplification protocol(TRAP) combined with silver staining in 42 low differentiated squamous carcinoma, 10 normal nasopharyngeal mucosae and 8 nasopharyngeal fibrohemangioma. RESULTS: Telomerase activity was detectable in 20% normal nasopharyngeal mucosae and 88.1% in NPC. The activity of telomerase was not detectable in 8 nasopharyngeal fibrohemangioma tissues. Telomerase activity was present more frequently in NPC biopsies with positive lymph node metastasis than in those without lymph node metastasis (P = 0.035). Furthermore, telomerase activity was observed more frequently in advanced NPC than in early stage (P = 0.048). However, there was no evidence to show the relationship between telomerase activity and other clinical features, such as age, gender and distant metastasis (P > 0.05). Serum EBV marker(VCA-IgA) was observed more frequently in NPC than in normal nasopharyngeal mucosae patients(P < 0.001). CONCLUSION: Telomerase activity may play a critical role in the tumorigenesis of NPC. Detection of telomerase activity combined with serological EBV markers may contribute to the screening and early diagnosis of NPC. PMID- 12768709 TI - [The study of p16 gene mutation and methylation in laryngeal squamous cell carcinoma]. AB - OBJECTIVE: To investigate the development and mechanism of the p16 gene inactivation in laryngeal squamous cell carcinoma (LSCC). METHODS: With PCR-based methylation assay and PCR-SSCP, first and second exon mutation in p16 gene and methylation of endogenase in promoter in 32 cases of LSCC and paracancer tissue were studied. RESULTS: No mutation were found in first and second exon in the samples; methylation of Cfo I were detected in 6 cases in first exon, methylation of Sac II was found in 4 cases, and methylation of Hpa II in 4 cases. In second exon, methylation of Hpa II was detected in 5 cases and methylation of Cfo I in 6 cases. CONCLUSION: Methylation of promoter in p16 gene is one of the important mechanisms of the inactivation of the gene. The inactivation of p16 gene has a close relation with the development and progress of LSCC. PMID- 12768710 TI - [Laryngeal reconstruction of laryngectomee]. AB - OBJECTIVE: To restore the laryngeal function of laryngectomee. METHODS: From 1990 to 1999, laryngeal reconstructions were performed in 34 cases of laryngectomee. The patients were 33 male, and one female. The age range was 27 to 77 years, with a mean age of 56.6. Thirty-three cases were total laryngectomized, one case due to laryngeal injury. The duration of alaryngeal was 2-72 months, with a mean of 18 months. The characteristic of this operation is that a cervical myocutaneous flap was used to link the pharynx and trachea, to act as an epiglottis. RESULTS: Speech function in all patients was good. One case can breathe through the larynx after decannulation and 13 cases can breath through the larynx in daytime. CONCLUSION: Laryngeal reconstruction for laryngectomee is applicable in laryngeal operation. PMID- 12768711 TI - [Pharyngeal plexus neurectomy for voice restoration of alaryngeal]. AB - OBJECTIVE: To release the spasm of pharyngeal constrictor and raise the successful rate of voice rehabilitation in total laryngectomized patients wearing Blom-Singer prosthesis. METHODS: The main branch of the pharyngeal plexus nerves within the inferior contractor can be found at the posterior side of hyoid greater cornu, which is about 6.5 mm from the back of constrictor. The spasm of the inferior constrictor can be resolved by cut off the nerve. RESULTS: Voice restoration was achieved in 15 cases after pharyngeal plexus neurectomy, the successful rate was 93.75%. CONCLUSION: It is feasible to replace pharyngeal myotomy by nerve plexus neurectomy and raise the voice rehabilitation rate. PMID- 12768712 TI - [Level choice of the neck dissection in clinical N0 patients with supraglottic cancer]. AB - OBJECTIVE: To study the characteristics of the cervical lymph node metastasis in negative node with clinically supraglottic cancer patients and decide which level should be dissected reasonably. METHODS: 57 modified neck dissection (MND) cases in the cN0 with supraglottic cancer were performed. Each lymph node in the 57 (63 sides) MND samples was studied pathologically in order to define the level of the lymph node metastasis. RESULTS: 1,877 lymph nodes were obtained in these MND samples; an average of 29.8 lymph nodes was obtained in one side. Among them 43 lymph nodes were metastasis nodes and 41 lymph nodes were located in the II, III level, 95.4% of metastasis cases. 15 cases (17 sides) in the 57 patients were metastasis (26.3%) and 14 cases of them were in the level of II and III (93.3%). Ipsilateral cervical recurrence occurred in the 3 of 57 cases in the II, III and IV level respectively. The recurrent rate was 5.3%, and 5-year survival rate was 80.7%. CONCLUSION: II and III levels should be dissected in the supraglottic cancer with cN0 and IV level should be included when III level was involved. I and V levels should not be included when the evidence of metastasis is not enough. PMID- 12768713 TI - [Surgical management of cervical esophageal carcinoma]. AB - OBJECTIVE: To review the experience of different surgical procedures and corresponding one-stage construction methods for cervical esophageal carcinoma. METHODS: 1. Thirty-three patients with cervical esophageal carcinoma were retrospectively reviewed, including 31 males and 2 females. According to UICC 1997 criteria and pathologic findings showing squamous cell carcinomas, there were 2 patients in T1N0, 4 in T2N0, 1 in T2N1, 6 in T3N0, 6 in T3N1, 7 in T4N0 and 7 in T4N1 categoris. 2. Removal of cervical esophagus was conducted in 13 patients. The type of cervical esophageal reconstruction included direct suturing, pectoralis major musculo-cutaneous flap or combined with the split graft, laryngotracheal flap and colon interposition. Total esophagectomy without thoracotomy was carried out in 20 patients. Reconstruction of the esophagus consisted of 17 stomach pulling-up and 3 colon interposition. Twenty-four patients received radiotherapy postoperatively(dose 50-70 Gy). RESULTS: The resectability of cervical esophageal carcinoma was 100%. The overall follow-up was from 4 months to 10 years. Two and one patient in T1, 2 survived 3 and 5 years, respectively. Three and five-year survival rates for T3, 4 were 7/16 and 3/8, respectively. Twenty-six patients had laryngeal function preserved and acquired good speech. The decannulation rate was 80% (20/25). The incidence of complication was 33.3%, including pharyngeal fistulas in 6 cases, cervical stomal fistulas in 2, cervical stomal stenosis in 2 and splitting of abdominal wall in 2. CONCLUSION: Surgical resection of cervical esophageal carcinoma and resolvation of the extraesophageal invasion is possible. The continuity of the esophagus is restored by pectoralis major muculocutaneous flap, laryngotracheal flap, stomach transposition and colon interposition. Combined with radiotherapy, the resectability and survival rate of the cervical esophageal carcinoma can be improved. PMID- 12768714 TI - [Effect of microwave coagulation and surgery to treat the cancer of the floor of mouth]. AB - OBJECTIVE: The aim of this article was to clarify the value of microwave coagulation to treat the cancer of the floor of mouth by comparing the effect of microwave and surgery. METHODS: Retrospectively 87 cases of cancer of the floor of mouth who were treated by microwave coagulation and traditional surgery are analysed and the survival curve, the recurrent rate of the primary and secondary site, the oral functions, the appearance and the complications are compared. RESULTS: 1. Either in the early patients (T1 + T2) or in the advanced cases (T3 + T4), the differences of survival curve between the two groups were insignificant (P = 0.8247, P = 0.803 respectively); 2. The recurrent rate of the primary and secondary site were insignificant either (P = 0.988, P = 0.759 respectively); 3. There were more patients with a satisfied mouth function in the microwave coagulation group than that of in the surgery group, on the contrary, the cases with a worst function were few in the former(P = 0.000, P = 0.001 respectively); 4. The appearance was more satisfactory in the microwave group(chi 2 = 23.077, P = 0.000). CONCLUSIONS: To treat the cancer of the floor of mouth, microwave coagulation, as the traditional surgery, had good effects such as the same survival rate, the complication rate and local or region control rate, but the mouth function and the appearance was better than the surgery group. The microwave coagulation can be safely carried out in the practice. PMID- 12768715 TI - [Election for surgery access to skull base]. AB - OBJECTIVE: To seek for a better approach for the resection of skull base tumors. METHODS: A review of 84 skull base tumors resection employed unlike operative approach in the Cancer Hospital of Chinese Academy of Medical Sciences was presented. Surgical approach feature of invaded the different position of skull base, selects the best operative access for safely removes the tumor, protects the critical structure of skull base as fully as possible, and decreases the complication were discussed in this article. RESULTS: Fourteen cases of benign and seventy cases of malignant underwent surgery. The tumor in 38 cases implicated parapharyngeal space and skull base. The lesion synchronously involved the anterior and middle cranial fossa in 7 cases, middle and posterior cranial fossa in 13 cases, lateral cranial base in 26 cases. The survival rate at 3 years was 68.5% (27/40) for malignant tumor. There was no operative death and severe complication in this series. CONCLUSION: To select the best operation access should think over following factors: 1. Tumor position and the scope implicated. 2. Safely and fully removal of the tumor. 3. The limited destroy to normal function structure and receivable cosmetic effect. 4. It is convenient to reliable reconstruction. PMID- 12768716 TI - [Clinical and pathological study of chordoma in the skull base]. AB - OBJECTIVE: To study the clinical and pathological characteristics of chordoma in the skull base. METHOD: The clinical manifestation, radiological finding and pathological characteristics in 20 patients with skull base chordoma were reviewed. Among them, 7 were immunohistochemical stained for keratin, epithelial membrane antigen, vimentin, glial fibrillary acidic protein and desmen, 2 were studied by electron microscope. 19 patients received surgical resection(5 cases were assisted by nasal endoscope). Eleven patients received radiation therapy. All 19 patients were followed-up for 3-5 years after treatment. RESULTS: (1) Clinical type were divided into four types in this group according to symptoms, sign, imaging features, operation finding, the original places and the expansive direction of the tumor. Seller type 5 cases, clival type 9 cases, occipito temproral type 2 cases and extent type 4 cases. One case with multiple cranial never palsies died due to the pressure on brain stem by the neoplasm, was autopsied prove lungs metastasis. 5 years survival rates were 65% and 35% respectively. (2) Vacuolated physaliferous cell is the pathology characteristic of chordoma. (3) Positive rate of epithelial marker antigens are higher than other mesenchymal neoplasms. Distended rough endoplasmic reticulum and filament were found within the cytoplasm under transmission electron microscope. CONCLUSION: Chordoma is a kind of low malignancy which express epithelial and mesenchymal characteristics. Vacuolated physaliferous cell is the characteristic Pathologic appearance. Immunohistiochemical stain is helpful in differential diagnosis. Complete excision is still technically impossible. Chordoma maintain high recurrence rate after surgery, with metastasis. Combined modality therapy include surgical operation and radiation could prolonged survival period. PMID- 12768717 TI - [Anatomic investigation of the labyrinthine artery]. AB - OBJECTIVE: To observe the origin, the course and the location of the labyrinthine artery[LA], and to investigate the spacial relationship between LA and facial nerve and vestibulocochlear nerve (VCN). METHODS: The specimens including 49 sides cerebellopontine angle (CPA) and internal acousticmeatus (IAM) infused with red galatin into artery were dissected and observed under operating microscope. RESULTS: LA arised from anterior inferior cerebellar artery (AICA) (83.6%), or basilar artery (BA) (12.3%) or vertebral artery (VA) (4.1%). AICA formed a loop in cerebellopontine angle (CPA) or internal acoustic meatus (IAM). The loop was located in the extra-meatus of IAM (28.6%), or at the opening of the IAM (18.4%), or in intrameatus of IAM (36.7%). AICA passed over ventral side of VII and VIII cranial nerves (18.4%) or across between VII and VIII cranial nerve root (81.6%). LA was a mono-arterial (51.1%), or bi-arterial (40.8%), or tri-arterial (4.1%) vessel. The calibre of LA was (0.18 +/- 0.05) mm. In CPA and IAM, facial nerve located anteroinferior to VIII cranial nerve, but the location of VCN was posteriolaterally. LA most commonly coursed between VII and VIII. LA divided into anterior vestibular artery, vestibulocochlar artery and cochlear artery. On the base of IAM, the facial nerve situated in anterosuperior, but cochlear nerve in anteroinferior and vestibular nerve in posterosuperior. CONCLUSION: LA was a mainly artery supplied to facial nerve, VCN and vestibulocochlear organs. Once LA is damaged, facial weakness (or paralysis), vertigo or hearing loss would be developed. PMID- 12768718 TI - [Preliminary study on inner ear ischemic model induced by ferromagnetic embolism]. AB - OBJECTIVE: To set up an inner ear ischemic model in guinea pig with ferromagnetic embolism. METHODS: A magnet was fitted in the external auditory canal and carbonyl iron filings (1%, 1 ml/kg) was injected into jugular, then the inner ear vessels were obstructed by ferromagnetic spheres. Cochlear blood flow (CBF) and number of red blood cells in the stria vessels were used to detect the model's ischemia of cochleae. The slice of temple bone and basal membrane stained by silver nitrate were used for inner ear's histopathological observations. RESULTS: The iron spheres were amassed in the one and two-day-later's model of inner ear vessels, which resulted in embolism. The number of red blood cells in the stria vessels decreased and then recovered to normal level after 4 days, but the CBF decreased to 50% +/- 10% of basic level immediately and recovered to 99% +/- 41% 4 days later. Scattered lesion of out hair cell cilium could be seen in cochleae in eight-day-later's model, and degenerations in different degree were found in vascular stria. CONCLUSION: The methods of inner ear ischemic model with ferromagnetic embolism could be practical and the decrease of CBF was reversible, so it may be an ideal model for studying some ischemic inner ear diseases and evaluating the effects of therapeutic drugs. PMID- 12768719 TI - [Protective effect of adenoviral-mediated brain derived neurotrophic factor gene on spiral ganglion]. AB - OBJECTIVE: To observe the expression of brain derived neurotrophic factor (BDNF) in the cochlea of guinea pig, and assess the activity of BDNF in spiral ganglion cell following the damage of noise. METHODS: Twenty-seven guinea pigs were exposed to a 4 kHz narrow band noise at 135 dB SPL for 4 hours. At seven days after the noise exposure, twelve guinea pigs were inoculated with ad-BDNF, and twelve guinea pigs were inoculated with ad-LacZ, while three guinea pigs were inoculated with artificial perilymphatic fluid. The animals were sacrificed After 1 week, 4 weeks and 8 weeks, respectively. The cochlea was stained with immunohistochemisty (ABC method) to examine the expression of BDNF. The number of spiral ganglion cell was counted to assess the activity of BDNF. RESULTS: BDNF gene was expressed in whole cochlea, and the expression was obviously elevated 1 week after the adenovirus administration and reduced thereafter. The number of degenerated cells in ad-BDNF group in the eight weeks was lower than that in another two groups and the different between them was statistical significant (P < 0.01). CONCLUSION: Adenoviral-mediated brain derived neurotrophic factor can be expressed in a high level in the cochlea of guinea pig, and may prevent the cell of spiral ganglion from the damage of noise. This study lays the groundwork for alleviation of hearing loss using gene therapy. PMID- 12768720 TI - [Experimental study on laryngeal cartilage defects repair with differently preserved allogeneic cartilages transplantation]. AB - OBJECTIVE: To study the effect of laryngeal cartilage defect repair with differently preserved allogeneic cartilage grafts. METHODS: 16 New Zealand white rabbits were used and divided into two groups. A 6 mm x 3 mm x 1 mm whole thickness cartilage defect was made in each side of the thyroid cartilage of each rabbit. In group one, the tissue-cultured cartilage grafts, preserved in RPMI 1640 medium for 30 days, were implanted in the left defects and the 4% formaldehyde preserved cartilage grafts for 30 days were implanted in the right defects. Fresh autogenous and allogeneic grafts were seprately transplanted into the right and left thyroid cartilage defects of the other group. Thyroid cartilages were taken out at 7, 30, 180 and 360 days after implantation. Samples were observed by macroscopy and prepared for hematoxylin and eosin (H&E) staining and immunohistochemical examination. RESULTS: No marked changes in form and volume were found in the fresh allografts and RPMI-1640 cultured cartilage grafts. The same as the autogenous cartilages, the defects of thyroid cartilage were successfully repaired by the fresh allografts and RPMI-1640 cultured cartilages. Whereas 4% formaldehyde preserved cartilage grafts were completely absorbed and replaced by cicatricial tissues in the thyroid cartilage defects. Histological observation showed that severe inflammatory cellular infiltration in the formaldehyde preserved cartilages at 7 and 30 days. The cartilage matrixes were resorpted and the chondrocytes showed degenerative change. Finally they were replaced by fibrous connective tissue at 360 days. In the RPMI-1640 cultured and fresh allogeneic cartilages only a little inflammatory cellular infiltration was observed and the cartilage matrixes were almost normal from 7 to 360 days. CONCLUSION: It is clinically feasible to use allogeneic grafts stored in RPMI 1640 medium or fresh allogeneic cartilage grafts for repairing laryngeal cartilage defects. PMID- 12768721 TI - [Chronic stenosed otitis externa: a report of 14 cases]. AB - OBJECTIVE: To evaluate the clinical features of chronic stenosed otitis externa (CSOE) and its treatment. METHODS: Fourteen patients with CSOE from 1992 to 2000 were analyzed retrospectively. RESULTS: In etiologic study, otitis externa and eczema were found in nine cases, chronic suppurative otitis media in three, acetic acid burn in one, and no apparent precipitating causes in one. Ten cases were cured through single operation, 3 experienced 2 operations and one had had 3 operations. The restenosis of external auditory canal occurred in four ears. CONCLUSIONS: CSOE is a rare disease. Surgical treatment is the main effective method for CSOE. The important items for treatment are: removing all stenotic fibrous tissue, widening bony canal, covering all bare bone with skin, preventing wound infection and close postoperative follow-up. PMID- 12768722 TI - [Immunohistochemical study of the epithelial hyperproliferation in middle ear cholesteatoma]. AB - OBJECTIVE: To elucidate the role of epidermal growth factor (EGF) in the growth of cholesteatoma, and to investigate the biological behaviour of cholesteatoma. METHODS: The expression of EGF receptor (EGFR), proliferating cell nuclear antigen Ki-67, collagen IV protein and laminin in 18 specimens of cholesteatoma was stained immunohistochemically and compared with that in epidermis of 8 specimens of normal external auditory meatus skin. RESULTS: The immunostaining for EGFR showed two patterns: strong linear or granular cell surface staining and granular cytoplasmic staining. In epidermis of normal skin, EGFR appear to be highly expressed on the basal layer. Cholesteatoma epithelium showed increased positive reactions in the super basal layers. The Ki67 was expressed in nuclei, and a dot-like as well as a diffuse nuclear staining pattern was detected. The Ki67 expression was restricted in the basal layer of normal epithelium, and the positive of Ki67 in nuclei was rare. The expression of Ki67 in cholesteatoma extended to the suprabasal and upper layers of the epithelium with an increasing frequency of positive cells. Collagen IV and Laminin were similary localized in the basement membrane of normal epithelium with a continuously staining pattern and a similar staining pattern was also found in the cholesteatoma, and the vas is abundant under basement membrane in some cholesteatoma samples. CONCLUSION: The overexpression of EGFR suggest a hyperproliferative state in cholesteatoma and there is a close relationship between the EGF and hyperproliferation of cholesteatoma. The increased expression of Ki67 confirms the hyperproliferative nature of cholesteatoma. The results of expression of Collagen IV and Laminin in cholesteatoma indicate it is a benign disease. PMID- 12768723 TI - [Expression of transforming growth factor-beta 1 matrix metalloproteinase-1 and its inhibitor in human middle ear cholesteatoma]. AB - OBJECTIVE: To investigate the expression of matrix metalloproteinase-1 (MMP-1), tissue inhibitor of metalloproteinase-1 (TIMP-1) and transforming growth factor beta 1 (TGF-beta 1) in human middle ear cholesteatoma. METHODS: Immunohistochemical staining was used to detect the expression of TGF-beta 1, MMP 1 and TIMP-1 in the cholesteatoma (41 cases) and the skin of external auditory meatus (15 cases). RESULTS: The percent of positive expression of TGF-beta 1, MMP 1 and TIMP-1 in cholesteatoma was 95.12%, 85.37% and 9.76%, respectively. There was significant difference of the expression of MMP-1 and TGF-beta 1 between cholesteatoma and external auditory meatus skin (P < 0.05). But there was no statistic significance for TIMP-1 (P > 0.05). There was correlation between the expression of TGF-beta 1 and MMP-1, but no correlation between TGF-beta 1 and TIMP-1. CONCLUSION: The imbalance between the expression of MMP-1 and TIMP-1 in cholesteatoma may cause directly proteolytic activity involved in pathologic osteolytic mechanisms. PMID- 12768724 TI - [Analysis of sever and rare complications of transsphenoidal surgery for lesions of sella turcica]. AB - OBJECTIVE: To analysis the reason and prevention of sever and rare complications of transsphenoidal surgery for lesions of sella turcica. METHODS: The clinical data of 16 cases who occurred sever and rare complications during transsphenoidal surgery for 200 cases of lesions of sella turcica was analyzed. The reason and prophylaxis of complications was inquired. RESULTS: The sever and rare complications occurred in 16 cases. 4 cases died and 8 cases bleed severly and the version of 2 cases was deteriorated and 2 cases had mental disorder and 2 cases had epilepticism and 1 cases had sever postoperative headache and the brain was damaged in 2 cases among 16 cases. CONCLUSION: Although the transsphenoidal surgery comparing with transcranial surgery has mini-invasive, many complications can occur and can be decreased by selecting surgical accommodation strictly and similarity with anatomy of sella turcica and mastering transsphenoidal surgical technique skilled. PMID- 12768725 TI - [Effect of erythromycin on apoptosis of eosinophils in nasal polyps in vitro]. AB - OBJECTIVE: To study the effect of erythromycin on apoptosis of eosinophils in nasal polyps in vitro. METHODS: Fifteen nasal polyp biopsies were collected during nasal polypectomy. The specimens were cultured with erythromycin in RPMI1640, and cultured without erythromycin as control group. The explants in both groups were taken out at 1, 3, 5 day after culture. The apoptosis of cells was detected by terminal deoxynucleotidyl transferasemediated dUTP nick endlabeling(TUNEL), eosinophils were identified by morphology and serial sections stained with HE, then the apoptosis index (AI) of eosinophils was calculated. RESULTS: The AI of eosinophils in nasal polyps after cultured for 1,3,5 days with erythromycin were (31.77 +/- 9.52)%, (32.67 +/- 9.44)% and (50.34 +/- 8.78)% respectively. In control group, the AI of eosinophils were (29.18 +/- 7.31)%, (29.82 +/- 12.03)% and (42.25 +/- 8.08)% respectively. There were significant difference between two groups at 5 day after culture(P < 0.05). CONCLUSION: Erythromycin promoted apoptosis of eosinophils in nasal polyps in vitro. PMID- 12768726 TI - [Bleeding control in sinonasal microsurgery]. AB - OBJECTIVE: To explore the results of bleeding control in sinonasal surgery. METHODS: Between Jan 1998 and Jun 2000, 82 patients (144 sides) were observed and studied for the relationship between mental tension and bleeding. The effect of blood pressure control and operative time reduction was also observed. 58 sides were of type III, 71 sides of type II (among them, 56 sides belonged to stage II, 15 sides belonged to stage III), 15 sides were type I (among them, 4 sides belonged to stage I, 11 sides belonged to stage II). Of them, 60 patients had been operated for two times. All patients with sinusitis received micro intranasal ethmoidectomy and 140 sides had had polyps removed. Sixteen patients received correction of deviated septum at the same time. 31 patients received resection of the inferior turbinate bone. Twenty-one patients were given Dridol' 5 mg and Dolantin 50 mg intravenously to control hypertension. Bleeding amount was compared between 21 hypertensive patients who used intravenous drugs and 57 hypertensive patients who did not use intravenous drugs during operation. To reduce operation time, the operation was simplified. RESULTS: The bleeding was limited to 10 ml in 99 sides(69%), the mean bleeding amount was controlled to (18.7 +/- 24.4) ml in 78(54%) sides of patients with hypertension. In 66 sides (46%) of patients with normal blood pressure, the mean bleeding amount was (13.2 +/- 16.8) ml (P < 0.05). In 21 patients in whom the hypertension was controlled, the mean bleeding amount was (13.0 +/- 6.7) ml. In 10 patients, the bleeding amount was (30.5 +/- 21.8) ml at the first operation with hypertension uncontrolled, the bleeding amount was only (11.0 +/- 8.2) ml at second operation when the blood pressure was controlled to normal (P < 0.05). After 6 months follow-up, 104 sides (72%) were cured, 32 sides (22%) improved, and 8 sides (6%) without improvement. CONCLUSION: Mental tension in operation can induce hypertension and increase bleeding, suitable sedation is necessary. With correct use of topical anesthesia and reduced operation time, the bleeding amount can be controlled to less than 10 ml in 69% of patients. PMID- 12768728 TI - [Study on the directed inducing process of cartilage cells differentiated from human marrow mesenchymal stem cells]. AB - OBJECTIVE: To study the feasibility of constructing tissue engineered cartilage cells by differentiated human bone marrow mesenchymal stem cells(MSC) cultured in vitro. METHODS: The MSC were isolated from the rib by density gradient centrifuge, then induced into chondrogenic differentiation by adding dexamethasone, transforming growth factor-beta 1 (TGF-beta 1), ascorbic acid, insulin and bovine serum albumin. RESULTS: Some cells were degenerated and disappeared after cultured in vitro. But the residual cells were surviving and have secreted metachromatic matrix, which is positive with toluidine blue staining Collagen II antigens expression is positive, which is one of the cartilage cells characteristics. CONCLUSION: MSC can be induced to cartilage cells in vitro, MSC can be used as functional cells for constructing tissue engineered cartilage. PMID- 12768727 TI - [Localization of upper airway stricture in patients with obstructive sleep apnea syndrome by CT scan]. AB - OBJECTIVE: To evaluate the diagnostic significance of CT scan in the localization of the stricture in the upper airway in patients with obstructive sleep apnea syndrome (OSAS). METHODS: Fifty-four patients with OSAS were included in this study. CT scan evaluated the upper airway from the roof of nasopharynx to the glottis using a Phlips Tomoscan AV Expander E1 spiral scanner. The areas and the dimensions of palate, uvula, lingua and epiglottis region, as well as the thickness of retropharyngeal and lateral pharyngeal tissue was evaluated. The reference values had been set-up in 225 normal adult upper airways CT scan, some patients' results were compared with the CT scan results during apnea. RESULTS: There were no any upper airway stricture in 12 patients with OSAS, and there were one or more upper airway stricture sites in other 42 patients. Twenty-four patients had only one stricture site. Fourteen patients had 2 adjacent stricture sites. One patient had three stricture sites. Three patients had 4 upper airway strictures. There was a good concordance between the results of CT scans during awakening and sleeping in 14 patients. CONCLUSION: Most patients with OSAS had anatomic upper airway strictures, the CT scan can localize the stricture site very well. PMID- 12768729 TI - [Evaluating effects of some medicine on tinnitus with animal behavioral model in rats]. AB - OBJECTIVE: To develop a valid behavioral animal model of tinnitus and to determine the animals whether have the perception of tinnitus and to evaluate effects of nimodipine and Chinese herbs on tinnitus using the behavioral model. METHODS: Adult male albino rats (n = 60) were fell evenly and randomly into ten groups. Animals in Group I and II were administrated sodium salicylate 350 mg/kg/d (i.p.). Instead, animals in control groups (Group III and IV) were saline injected. Pure tone (8 kHz, 65 dBSPL) was given to rats in Group V and VI instead of sodium salicylate. Nimodipine (1 mg/kg) were administrated to animals in Group VII and VIII. Gushenshuerfang, one kind of Chinese herbs were administrated to animals in Group IX and X. Medicine or pure tone were being given to all animals in Group I, III, V, VII, IX before conditioned reflex. Animals in Group II, IV, VI, VIII, X began to have medicine or pure tone after conditioned reflex. Thirsty animals (deprived of water for several days) were breeded in sound-proofed room and listened a persistent white noise (55 dB SPL) and trained daily with our instrument system, according to the procedure based on Pavlovian conditioned suppression paradigm. Subjects were permitted to lick water at an automatic dispenser during daily training sessions and the number of licks was recorded automatically. Noise offset (silence) was the conditioned stimulus and the electrical shock was the unconditioned stimulus. After two or three days training, subjects learnt rapidly to associate noise offset with shock and then established a behavioral conditioned reflex. Because of shock fear they decrease or even stop licking with the presentations of conditioned stimulus. However, subjects gradually forgot or ignored the fear without any shock present in extinction test sessions. RESULTS: Animals' licking recovered to their pre training levels respectively at the following days: Group I--5 days, Group II--2 days, both Group III and IV--4 days, Group V--5 days, Group VI--2 days, Group VII -5 days, Group VIII--4 days, Group IX--5 days, Group X--4 days. There were much significant differences among all groups and all training days (F test, P < 0.001). Above results provided sufficient evidence for that tinnitus was induced by salicylate and tinnitus presented with noise offset. In group I, tinnitus became additional conditioned stimulus. Therefore, the fear of animals was much severe and the numbers of licking were much fewer. Extinction lasted longer. In group II, animals ignored the noise offset much easily and the extinction lasted much shorter. Moreover, results from group V and VI, pure tone instead of salicylate, indicated that sensation (tinnitus) sounded like pure tone. The groups VII, VIII, IX, X demonstrated that nimodipine and Gushenshuerfang, one kind of Chinese herbs can cure the tinnitus induced by salicylate. CONCLUSION: All behavioral evidence indicated that animals perceived tinnitus induced by salicylate and this animal model could be used in evaluating curative effects of some medicine on tinnitus. PMID- 12768730 TI - [Establishment of the animal model of triple semicircular canal occlusion]. AB - OBJECTIVE: To establish the animal model of triple semicircular canal occlusion(TCO) in guinea pigs. METHODS: TCO was performed on 20 guinea pigs. Electronystagmography, auditory brainstem responses (ABR) and distortion product otoacoustic emissions(DPOAE) were recorded pre- and post-operatively. The non operated ear was used as control. RESULTS: There was spontaneous nystagmus towards non-operated side on the 1st postoperative day and no nystagmus to stimuli of sinusoidal angular acceleration in operated side. On the 30th postoperative day, left and right nystagmus returned symmetrical. Pre-operation, ice-water stimulation induced normal symmetric nystagmus in both sides. On the 30th postoperative day, ice-water stimulation only induced normal nystagmus in non-operated ear and failed in operated one. ABR thresholds increased approximately 10 dB post-operation. There were no significant changes of the amplitudes of DPOAE before and after surgery. Histologic examination revealed complete occlusion in the three semicircular canals with newly formed bone and connective tissue. CONCLUSION: This study showed that the animal model of TCO was successfully established. These findings suggests that TCO might be a safe and simple procedure for the treatment of intractable peripheral vertigo. PMID- 12768731 TI - [Effects of microcirculatory disorders of inner ear on blood-labyrinth barrier permeability in guinea pigs]. AB - OBJECTIVE: To study the changes of blood-labyrinth barrier permeability during cochlea microcirculatory disorders in guinea pigs. METHODS: Using modified method of Evan's blue fluorescence, the changes of permeability of blood-labyrinth barrier were observed in the animal model of cochlea microcirculatory disorders which was induced by photochemical reaction. RESULTS: Amount of Evan's blue passing through the blood-labyrinth barrier was (1.709 +/- 0.769) microgram per guinea pig after 2 hours and (2.849 +/- 0.653) microgram per guinea pig after 4 hours when the cochlea was in microcirculatory disorder. CONCLUSION: The results indicated that the permeability of blood-labyrinth barrier increased when cochlea microcirculatory disorders occurred and the increase of blood-labyrinth barrier permeability may be one of the important mechanisms which causes the cochlear ischemic lesions. PMID- 12768732 TI - [Expression of simian virus 40 large T-antigen and p53 protein in chinchilla middle ear epithelial cell line and its significance]. AB - OBJECTIVE: To identify the expression of simian virus 40 large T-antigen (SV40TAg) and p53 protein in the cell nuclei of chinchilla middle ear epithelial cell line and explore the significance of the expression. METHODS: To examine the expression of SV40TAg and p53 protein in cell nuclei of chinchilla middle ear epithelial cell which had been infected and had not been infected with Ad 12-SV40 hybrid virus by immunocytochemistry method. RESULTS: All of the cell nuclei in 1 of 8 culturing dishes which infected with the Ad12-SV40 virus expressed the SV40TAg and p53 protein and had cultured over 46 passages. They have become a cell line. All of the cells in rest 7 culturing dishes and the control cells did not express the SV40TAg and p53 protein in their nuclei and died after culturing 6-8 passages. CONCLUSION: The SV40TAg and p53 protein play some important role in process of cell immortalization. The expression of SV40TAg and p53 protein in cell nuclei of CMEE-1 cell line has close interrelations with establishing the CMEE-1 cell line. PMID- 12768733 TI - [Expression of apoptotic gene and FGF on olfactory bulb of rat and its significance]. AB - OBJECTIVE: To study the expression and distribution of apoptotic gene Bcl-2 and Bax on olfactory of rat, and its relation with the FGF and to investigate the mechanism of senile dysosmia. METHODS: Ten young (3 months) and ten senile rats (24 months) were used in this study. After the removal of the heads of these rats, olfactory bulb were immediately fixed with neutralized formalin, followed with paraffin-embedding, serial sectioning, immunohistochemiscal staining and microscopic observing. RESULTS: The positive expression rate of Bcl-2 and FGF on olfactory bulb in young rats was significantly stronger than that in senile rats (P < 0.01). The positive expression rate of Bax in senile rats was a little stronger than that in young rats, but the differences were not statistically significant. (P > 0.05) CONCLUSION: The expression of FGF and anti-apoptotic gene Bcl-2 on olfactory bulb decrease with the increase of age. This suggests neuron apoptosis on olfactory bulb are regulated by neurotrophic factor, and directly related with senile dysosmia. The mitral cell apoptosis as a result of FGF decreased on olfactory bulb may play a key role in the senile dysosmia. PMID- 12768734 TI - [Jugulotympanic glomus tumors: a report of 11 cases]. AB - OBJECTIVE: To evaluate the surgical technique in removal of jugulotympanic glomus tumor. METHODS: Retrospective analysis of surgical techniques and effects of the surgical treated 11 cases (1982-1998) of jugulotympanic glomus tumors (tympanic type I, 1, II, 1, IV, 3; jugular type I, 1, III, 5). RESULTS: The tumors of all 11 patients were removed completely and the wounds healed smoothly. Only one case had postoperative laryngeal nerve paralysis. There was no recurrence after 1-8 years follow-up. CONCLUSION: Suitable surgical techniques are available for removal of jugulotympanic glomus tumors and are chosen according to the size and location of the tumor. Postauricular incision is suitable for Glomous tympanicum Type II, III, IV. Since postauricular incision with superior and inferior extension provides a good exposure of the neck and temporal bone, it is suitable for surgical removal of Glomus jugulare type I. In case of Glomus jugulare type III, postauricular large "C" incision may be chosen for resection of the mid- and infra-temporal fosse, neck and skull base tumor. PMID- 12768735 TI - [Expression of human papillomavirus DNA in cholesteatoma of the middle ear]. AB - OBJECTIVE: To reveal the role of HPV in the occurrence and development of cholesteatoma of the middle ear. METHOD: PCR and nuclear acid hybridization were applied in screening 44 cases of middle ear cholesteatoma for detection of HPV DNA, and a comparison analysis with the pathological results of 35 cases of middle ear cholesteatoma was made. RESULTS: Pathologically, the characteristics of HPV induced-lesions was found in 12 of 35 specimens (34.3%). 44 cases of middle ear cholesteatoma were examined with consensus primers PCR and digoxigenin labeled HPV general probes. HPV DNA positive rates were 29.5% (13/44) and 25.0% (11/44) respectively. HPV DNA positive rate was 58.3% (7/12) and 13.0% (3/23) respectively in 12 cases with pathological characteristics of HPV induced-lesions and in 23 cases without those characteristics. There was significant difference(chi 2 = 7.926, P < 0.005). CONCLUSIONS: HPV infection can arouse the cleavage and proliferation of cholesteatoma epithelium and may play certain role in the occurrence and development of middle ear cholesteatoma. Aggressive, papillomatous growth and koilocytes can be served as the initial proof of HPV infection in cholesteatoma of the middle ear. PMID- 12768736 TI - [A reappraisal of hyperbaric oxygenation effect and study on serum malondialdehyde and superoxide dismutase in patients with sudden deafness]. AB - OBJECTIVE: To reappraise the effect of HBO and to observe the changes of malondialdehyde (MDA) and superoxide dismutase (SOD) in serum of the patients with sudden deafness. METHODS: 60 patients were randomly divided into two groups. One was treated with HBO combined with medical treatment (treatment group). The other was only treated with HBO (control group). There were 30 healthy volunteers as health group. RESULTS: After treatment, there was a significant difference (chi 2 = 6.48, P < 0.05) in patient's groups in hearing examination. After treatment, SOD volume in treatment group had significant difference with health group and control group (t = 23.27, P < 0.01; t = 27.17, P < 0.01) respectively. There was a significant difference of SOD volume in treatment group itself before and after treatment course (t = 29.27, P < 0.01). There was no significant difference of SOD volume in control group itself before and after treatment(t = 1.33, P > 0.05). MDA: There was a significant difference between treatment group and control one after the treatment. There was a significant difference (t = 7.28, P < 0.01) in treatment group before and after treatment course. There was not a significant difference (t = 1.24, P > 0.05) between treatment and health group after the treatment course. There was a significant difference (t = 7.09, P < 0.01) between control group and health one after the treatment course. CONCLUSION: There was no satisfied curative effect on sudden deafness with HBO only. PMID- 12768737 TI - [Primary study of Na+ channels in the epithelial cells of nasal polyps and of the effect of IL-1 beta on Na+ channels]. AB - OBJECTIVE: To investigate the relationship between Na+ channels and nasal polyps and to study the effect of IL-1 beta on Na+ channels of epithelial cells of human normal nasal turbinate (NT) and nasal polyps (NP). METHODS: Serum-free cultured epithelial cells of NT from obstructive sleep apnea syndrome patients and NP were studied on collagen gel-coated membranes at an air-liquid interface using the patch-clamp technique in the cell-attached configuration. RESULTS: (1) The three Na+ absorption-related factors(the incidence of Na+ channels, the conductance of Na+ channels and the open probability of Na+ channels) were compared in NT cells and NP cells. There was no difference between NT and NP cells in the incidence(P > 0.05) and in the conductance(P > 0.05). However, the open probability was higher in NP cells compared to NT cells(P < 0.01). (2) Application of IL-1 beta increased significantly in the open probability of Na+ channels in NT and in NP cells (P < 0.01), the conductance of Na+ channels in NP cells was strongly increased by IL-1 beta(100 ng/ml)(P < 0.01). CONCLUSIONS: (1) The formation of NP is significantly correlated with increased Na+ absorption, the prolonged open time of Na+ channels is one of the most important factors. (2) IL-1 beta may upregulate Na+ permeability by prolonged open time of Na+ channels and increased conductance of Na+ channels, increased absorption of Na+ may lead to the growth and enlargement of NP. PMID- 12768738 TI - [T-lymphocyte subsets and inflammatory cytokines of interleukin-5 and interleukin 10 expression in human nasal polyp tissue]. AB - OBJECTIVE: To investigate the distribution of T-lymphocyte subsets (CD4+ and CD8+ cells) and the expression of IL-5 and IL-10 protein-positive cells in nasal polyps. METHODS: The specimens of thirty-two nasal polyps and six control subjects were studied by Immunohistochemical method to detect CD4+, CD8+, IL-5+ and IL-10+ cells. A mean numbers of positive cells per high power field (10 x 40) were counted. All data were analyzed with t-test and correl-test. RESULTS: There were significantly large number of CD4+ and CD8+ cells in nasal polyps (P < 0.001). A significant predominance of CD4+ cells over CD8+ cells was found in the two subgroups (subepithelia, arounding glands and blood vessels) of nasal polyp (P < 0.001). There were an abundant number of IL-5+ IL-10+ cells, whereas significantly difference higher concentrations than that in the controls (P < 0.001). IL-5+ cells were more than IL-10+ cells in nasal polyps (P < 0.05), and most of IL-5+ cells were plasmacytes and lymphocytes. There was a significant correlation between the counts of CD4+/CD8+ and IL-5+/IL-10+ in the subgroup of nasal polyps arounding glands and blood vessels (r = 0.78, P < 0.01). CONCLUSIONS: This study indicated that CD4+ T-lymphocytes and IL-5 high expression may play an important role in the nasal polyps. Migration of plasmacytes and lymphocytes in polyps may participate in the inflammatory reaction. The disturbance of cellular and humoral immunity and imbalance of cytokines distribution may play a crucial role in the pathogenesis of nasal polyps. PMID- 12768739 TI - [Endoscopic transnasal sphenoidal approach in hypophysectomy]. AB - OBJECTIVE: To report the anesthetization and surgical method of transnasal sphenoidal endoscopic approach in hypophysectomy. METHODS: Forty-two cases with pituitary tumor underwent tranasal-sphenoidal endoscopic surgery and 37 out of 42 accepted local anesthesia and sedation. RESULTS: No complications had occurred, the operations went successfully with less blood loss. Postoperative endoscopic follow-up period was performed at 6 to 35 months. The symptoms were improved in all patients. CONCLUSION: Our results show that transnasal-sphenoidal endoscopic approach under local anesthesia in the treatment of pituitary tumor seems to be a safe simple and valuable way. PMID- 12768740 TI - [Trannasal-transsphenoidal endoscopic surgery of the sphenoid sinus and the sella turcica]. AB - OBJECTIVE: The primary objective of this study is reporting our experience with transnasal endoscopic technique to management of the sphenoidal and the sellar lesions. METHOD: Forty one patients were management under endoscope who suffered from pituitary adenomas, craniopharyngioma and sphenoidal lesions, et al. RESULTS: In forty one cases of the sphenoidal and the sellar lesions, thirty two surgeries were successful. One patient died of bleeding, five developed the foramen of nasal septum, the adhesion of nasal cavity was complicated in three patients. CONCLUSION: The endoscope has the advantage of improved visualization, angled view, and a wide panoramic perspective. The transnasal-transspheniodal endoscopic technique is a effective, minimally invasive approach. But there are its disadvantages and occurred severe complication. Surgeons should be understood its feasibility, limits and indication. PMID- 12768741 TI - [The clinical study of tongue flaps repairing after resecting pharyngeal neoplasm and laryngeal neoplasm]. AB - OBJECTIVE: To study the repair methods of the defect after resecting pharyngeal neoplasm and laryngeal neoplasm. METHODS: Four kinds of tongue flap, such as 1/3 tongue flap, 1/2 tongue base flap, tongue base flap and transverse tongue flap were devised and applied in 15 patients with pharyngeal neoplasm and laryngeal neoplasm. RESULTS: These tongue flaps were alive in all patients. The wound of 13 patients healed in I stage. Two patients dehiscenced because of infection and healed after treatment. All the patients deglutited well. One-year, 3-year, disease-free survive rate were 92.9% (13/14), 72.7% (8/11) respectively. CONCLUSION: Tongue flaps are obtained easily, with enough blood flow, adapted to pharyngeal environment, easily alive and low complication in repairing the pharyngeal defect. We think that these tongue flaps should be applied in clinic. PMID- 12768742 TI - [Long-term follow-up result of partial laryngectomy]. AB - OBJECTIVE: To study the long-term follow-up result of partial laryngectomy and reservation of laryngeal function. METHODS: Three hundred and seventy-nine patients who underwent partial laryngectomy from 1986 to 1995 were summarized (male 290 cases, female 89 cases). Among them, 184 cases were supraglottic carcinomas (T1 8 cases, T2 115, T3 48, T4 13, according to UICC in 1992), 192 cases were glottic carcinomas (T1 115, T2 63, T3 13, T4 1), 3 cases were transglottic carcinomas (T2 1, T3 2). In common 8 kinds of operations were performed: 26 cases underwent cordectomy, 138 vertical laryngectomy, 7 frontolateral laryngectomy, 12 horizontal glottic laryngectomy (middle part of the larynx), 58 supraglottic laryngectomy, 95 horizontovertical (3/4) laryngectomy, 24 subtotal laryngectomy with cricoglossoepiglottic anastomosis, 19 near-total laryngectomy with cricoglossal anastomosis (with reservation of unilateral arytenoid cartilage). 193 cases underwent concurrent neck dissection(121 unilateral, 72 bilateral). RESULTS: All cases restored their phonation and overcame aspiration with removing nasal feeding from 7 to 23 days after operations. 362 cases were decannulated from 9 days to 3 months after operations. Another 8 cases were decannulated after a secondary plastic operation. Decannulation rate was 97.6%. The three, five and ten year survival rates were 86.8% (329/379), 81.3% (266/327) and 69.4% (120/173) respectively. CONCLUSION: Partial laryngectomy is a kind of radical operation with reservation of laryngeal function. Mastering indications strictly, correct operation choices, excellent surgical skills and perfect repairing technique are bases of improving life qualities and curative effect. PMID- 12768743 TI - [The clinical observation of microsurgery treatment in early glottic carcinomas]. AB - OBJECTIVE: To improve the life quality of the patients with T1 glottic carcinomas, and obtain good vocal effect. METHODS: Twenty-two patients under shoring laryngoscope and microscope were operated by surgery and laser. The phonetic parameters and vocal cord states of preoperative and postoperative patients were evaluated and analyzed objectively. RESULTS: The three-year relapse rate was 9.0% (2/22), and the survival rate of three-year, four-year, five-year were 100%, 94.7%, 93.3% respectively. The postoperative phonetic quality was improved. CONCLUSION: Patients with early glottis carcinomas could be treated by microsurgery. The damage of normal tissue might be reduced, and the patient's life quality has been improved. This method should be used widely in clinic. PMID- 12768744 TI - [Clinical characteristics of different cordectomy types with potassium titanium phosphate laser]. AB - OBJECTIVE: To study the clinical characteristics of different potassium titanium phosphate(KTP) laser types and surgical the recuperating course of cordectomy under suspension laryngoscope. METHOD: Follow-up observation were applied on 76 patients with cordectomy. Restoration of laryngeal structure and change of the quality of voice were observed and recorded. RESULTS: Type 1, normal laryngeal structure and function were restored postoperatively after 3 months; Type 2, a fold like vocal cord (new vocal cord) appeared 3 months later. Laryngeal structure was basically restored; Type 3, a mucosal fold appeared postoperatively after 6 months in most patients, but it lost motion function. Recurrence time was mainly found 2-4 months. Recurrence rate was 6.7%. Two and 5-year survival rate were 100%. CONCLUSIONS: Cordectomy with KTP laser can result in good effect and less injury. Type 2, 3 can retain thyroarytenoid muscle as much as possible on the basis of radical cure. PMID- 12768745 TI - [The relationship between tracheotomy and intra-tracheal papilloma progression in children]. AB - OBJECTIVE: To clarify the relationship between tracheotomy and intra-tracheal papilloma progression and introduce the experience of managing dyspnea in the hospital. METHODS: Children who were treated in ENT department of Tongren Hospital between 1992 and 1997 were reviewed in this essay. There were totally sixty-two children (thirty-two boys and thirty girls) with laryngeal papilloma and recurrent respiratory papillomatosis enroled in the series. RESULTS: Among 33 non-tracheotomied children the intra-tracheal lesions were found in only 2 cases (6.1%). There were 17 (80.9%) cases presented intra-tracheal lesions in 21 tracheotomied cases, and it showed great statistical difference (chi 2 = 27.4, P < 0.005). There were no difference of onset age, sex and the first sites of papilloma lesions between the non-tracheotomied and tracheotomied groups(P > 0.05). CONCLUSION: We found papilloma has a week potential to progress into trachea. Tracheotomy is the most important factor of intra-tracheal papilloma progression in many cases. The tracheotomy can be avoided. PMID- 12768746 TI - [Histological study of the local-regional invasion of pyriform sinus carcinoma]. AB - OBJECTIVE: In order to offer the basis for the operation of the pyriform sinus carcinoma, the local-regional invasion patterns of the pyriform sinus carcinoma were investigated. METHODS: Twenty-six surgical specimens obtained from patients who undergone laryngopharyngectomy were subjected to a whole organ section study. RESULTS: Tumors located in the lateral wall of the pyriform sinus spread mainly towards the lateral wall of the hypopharynx. Tumors located in the inner wall of the pyriform sinus spread mainly towards the larynx and opposite side pyriform sinus, the paraglottic space and thyroid cartilage were particularly frequent invaded. It was rare of the invading of the cricoid cartilage. The invasion of the epiglottis and preepiglottic space across the midline had not been found. There were two invading routines, the paraglottic space and preepiglottic space. The invasion of the paraglottic space was through the aryepiglottic fold and the inner surface of the thyroid cartilage. The invading of the preepiglottic space was through the aryepiglottic fold and the upper part of the inner surface of the thyroid cartilage. CONCLUSIONS: The invading of the preepiglottic space was not the contraindication of the partial laryngectomy. It is practicable to preserve the laryngeal function in most of the patients while tumors located on the lateral or inner wall of the pyriform sinus. There was a tendency to the invading to the contralateral side in the inner wall of the pyriform or postcricoidarytenoid region. Attention should be paid to the submucosal spreading in the postcricoidarytenoid region. PMID- 12768747 TI - [Precise map of allelic loss on chromosome 3p14 in nasopharyngeal carcinoma]. AB - OBJECTIVE: To determine the precise allelic loss on chromosome 3p14 and discuss the possible relations between loss of heterozygosity (LOH) and EBV infection, clinical stage and clinic-pathology of nasopharyngeal carcinoma (NPC). METHODS: Six high dense microsatellite marker on chromosome 3p14 were selected to examine LOH in 32 cases of NPC. RESULTS: 23 of 32 (71.9%) tumors were deleted for at least one locus of six loci. High frequencies of LOH (> 40%) were observed at loci D3S1300(50.0%), D3S1313(46.4%) and D3S1312(44.4%). 12 cases showed LOH in one contiguous and nonrandom region. The smallest common deletion region seems likely to lie between D3S1313 and D3S1312. Relations between LOH on 3p14 and clinical stage and EBV infection were observed. The frequency of LOH was 70.0% in 30 cases of poor-differentiated squamous cell carcinoma. 2 cases of vesicular nucleus cell carcinoma had LOH at two loci. CONCLUSION: The high deletion rate on 3p14 in NPC indicates that there might be a putative tumor suppressor gene related to the development and progression of NPC. PMID- 12768748 TI - [Whole genome-wide scanning for a large pedigree with matrilineal deafness]. AB - OBJECTIVE: To search responsible nuclear genes for matrilineal non-syndromic deafness. METHODS: Whole genome-wide scanning was performed to analyze the 365 short tandem repeats in a deaf pedigree with maternal inheritance using DNA pooling strategy. Frequencies of allele in one patient pool were compared with that in one unaffected relative pool as well as in one normal control pool. Linkage analysis was also conducted in some positive loci. RESULTS: Allele from 45 loci occurred more frequently in the patient pool than that in the unaffected relative pool and the control pool. No linkage was found from any candidate locus investigated. CONCLUSION: 45 positive loci obtained from this family may be as the candidate positions for matrilineal non-syndromic deafness. PMID- 12768749 TI - [Mitochondrial DNA4977 deletions associated with human presbycusis]. AB - OBJECTIVE: To determine whether or not the mtDNA4977 deletions contribute to human aging and involve in the development of presbycusis. METHODS: 67 sides of archival temporal bone sections, 20 temporal brains, 21 cochlear nucleus, 20 hearts and 22 livers were harvested and the total DNA was extracted. The presence of mtDNA4977 deletions was examined using nest polymerase chain reaction(PCR). RESULTS: Our results showed: (1) the highly conserved mitochondrially-encoded tRNA and ND1 segments were amplified from all tissues, as well as mtDNA4977 deletions related to aging. (2) The incidence of carrying mtDNA4977 deletions in all tissues in aged group was significant higher than that of young control group (P < 0.05). (3) The incidence of carrying mtDNA4977 deletions in temporal bone sections and cochlear nucleus with presbycusis patients was significant higher than that of aged normal hearing control group (P < 0.05). However, the incidence of carrying mtDNA4977 deletions in heart and liver were no significant difference between presbycusis patients and the control group. CONCLUSION: These findings indicated that mtDNA4977 deletions in human contribute to presbycusis, as well as aging. PMID- 12768750 TI - [Positron emission tomography of tinnitus-related brain areas]. AB - OBJECTIVE: To map the tinnitus specific central activity and foci site in brain and to investigate the effects of hearing loss, tinnitus side, right or left handed on foci site. METHODS: Glucose metabolic activity of brain was studied with positron emission tomography (PET) using 18F-FDG as radiotracer. Seventeen patients with severe tinnitus and fifteen control subjects participated in this study. All subjects were fell into four groups according to their hearing levels. There were 13 tinnitus patients with hearing loss in Group one, 4 tinnitus patients without hearing loss in Group two, 2 control subjects with hearing loss in Group three and 13 control subjects without hearing loss in Group four. Statistical parameters mapping (SPM) was used to analyze the PET data and to determine the Brodmann area (BA) according to Talairach coordinate system. RESULTS: Increase of neuronal activity caused by tinnitus occurred predominately in the left hemisphere with significant foci in the transverse temporal gyrus (BA41), superior temporal gyrus (BA42, 22), anterior middle temporal gyrus (BA38) and hippocampus. Bilateral hemispheres were concerned with the neuronal activity caused by hearing loss while foci site were located at the posterior superior temporal gyrus(BA42,22), medial portion of middle temporal gyrus(BA21), combined auditory area(BA39). Left middle frontal gyrus (BA8, 9) as well as left inferior frontal gyrus(BA45) were also link up with hearing loss. CONCLUSION: As a new path in research, PET has provided an objective evidence for tinnitus and may be used as a potential tool in objectively measuring tinnitus. Tinnitus-related brain areas and their influence factors were discussed. PMID- 12768751 TI - [The use of muscle autograft or nerve allograft denatured by microwave for repair of gaps in rat facial nerve]. AB - OBJECTIVE: Attempt to enhance recovery of rat facial nerve through different grafts. METHODS: Rats facial nerve mandible branch gap was repaired using different grafts, either muscle autograft or sural nerve allograft denatured by microwave. Axonal regeneration was studied in 10th week after insertion of the denatured muscle autografts or sural nerve allografts and compared with results found in autologous sural nerve grafts used as controls. Axonal regeneration, Schwann cell behavior and efficacy of nerve and muscle were quantified using CB HRP retrograde trace, HE staining, Flamming staining and electromyography. RESULTS: Denatured nerve allografts and muscle autografts supported the higher rates and volumes of axonal regeneration. Nerve allografts had the higher degree of myelin sheath developing. CONCLUSION: Nerve autograft and muscle autograft denatured by microwave are convenient, source sufficient, higher efficient grafts for repairing facial nerve gap and have potential clinical use. PMID- 12768752 TI - [Expression of cell proliferation and apoptosis gene associated protein on nasal polyps and its significance]. AB - OBJECTIVE: To further understand the pathogenic mechanism of nasal polyps, namely the cells proliferation and apoptosis in nasal polyps tissue. METHODS: The proliferation cell nuclear antigen (PCNA), apoptosis associated gene protein (Bcl 2, Bax) were determined in 26 tissue samples of nasal polyps and 14 controls from normal inferior turbinates respectively. RESULTS: (1) The positive expression e of PCNA, Bcl-2 and Bax were significantly higher in epithelium of nasal polyps than the controls, and the expression rate of Bcl-2/Bax, however, tended equipoise. (2) The expression of Bcl-2 were significantly stronger than Bax in glands, eosinophils of nasal polyps tissue, and there were not significant differences in inferior turbinates. CONCLUSION: There is strongly proliferation activity in the epithelium of nasal polyps, and expression imbalance of Bcl-2/Bax may be one of the important factors of eosinophilia in nasal polyps tissue. PMID- 12768753 TI - [Effects of nitric oxide on ciliary beat frequency in the human nasal mucosa]. AB - OBJECTIVE: To investigate the effect of nitric oxide on ciliary beat frequency in human nasal mucosa. METHOD: A primary culture model of human nasal mucosa epithelial cells was established and the effect of nitric oxide on ciliary beat frequency was observed by videomicroscopy method. The data were statistically analyzed using t-test. RESULT: L-arginine significantly increased ciliary beat frequency from (400 +/- 48) beats/min to (793 +/- 68) beats/min (n = 9, P < 0.001), D-arginine had no similar effect, while NOS blocker L-NAME inhibited the effect of L-arginine. CONCLUSION: Cells cultured by our method could synthesize NOS that made L-arginine produce NO that could increase ciliary beat frequency in the human nasal mucosa. PMID- 12768754 TI - [Investigation of the nasal T/NK cell lymphomas in 14 cases]. AB - OBJECTIVE: To study the biologic behavior, pathologic features as well as clinic manifestations of the nasal T/NK cell lymphomas, and to provide experiences for the diagnosis and treatment of the tumor. METHODS: Specimens of 23 cases lethal midline granulomas (LMG) were examined: (1) HE stain for immunohistochemical staining for TIA-1, CD43, CD20, CD3 epsilon, CD56 and in situ hybridization for EBV encoded small nuclear RNA(EBER1/2, Y017, DAK0). (2) The cases histories were reviewed. RESULTS: (1) 14 cases were confirmed by histological features. In the 14 cases, tumor cells expressed TIA-1, CD56, CD3 epsilon, CD43, EBER1/2 positive. (2) Among them 8 cases (57%) showed special clinic features of LMG, Nasopharynx, pharynx and laryngeal involvement were found in other 6 cases. One case revealed lymphoid leukemia. In other 2 cases, gastrointestinal tract were involved. Hemophagocytic syndrome (HPS) complication happened in 1 case. CONCLUSION: The tumor cells of nasal T/NK cell lymphomas expressed TIA-1, CD56, CD43, CD3 epsilon may suggest it originates from NK cell. The high association with EBV may suggest in situ hybridization. 57% patients showed that the special clinic features of the LMG, and gastrointestinal tract, bone marrow, peripheral blood were involved in some cases. One case complicated with hemophagocytic syndrome (HPS). PMID- 12768755 TI - [Cell proliferation in endolymphatic sac during the immune response of inner ear]. AB - OBJECTIVE: To study the cell proliferation in endolymphatic sac(ES) during the secondary immune response in inner ear. METHODS: Fifteen healthy, female SD rats were employed in the experiment. Sensitized systematically with keyhole limpet hemocyanin (KLH), the animals were inoculated with the same antigen through cochlea basal turn into the labyrinth. Administrated 5-bromo-2'-deoxyuridine (BrdUrd) intraperitoneally, the rats were sacrificed and the temporal bones were harvested at 3, 7, 14 day after labyrinth vaccination respectively. The frozen sections of the decalcified samples were dealt with H-E staining and immunohistochemical methods to investigate the cellular infiltration, BrdUrd and IgG positive cells in the ES. RESULTS: While the ES lumen and perisaccular region were infiltrated with mononuclear-phagocyte at the 3rd day post-vaccination, plasmocyte and lymphatic cells become the predominant infiltrating cells in the ES at the 7th day. The KLH in the ES lumen were phagocytized at the 3-7th day post-vaccination. S-phase cells and IgG positive cells in ES increased markedly in the 3rd day and 7-14 days post-vaccination respectively with similar localization. CONCLUSION: The activity of cell proliferation in the ES enhance and local proliferated cells may differentiate in situ into immunocompetent cells during the secondary immune response against TD antigen in the inner ear. This indicates that ES plays an important role in immune response of inner ear. PMID- 12768756 TI - [Central low frequency hearing loss]. AB - OBJECTIVE: To study the pathogenesis of the bilateral central low frequency hearing loss. METHODS: Audiologic findings [auditory brainstem response(ABR), evoked otoacoustic emission (EOAE), et al] of 101 cases with bilateral central low frequency hearing loss were studied. RESULTS: The typical clinical manifestations of bilateral central low frequency hearing loss were normal EOAE which cannot be suppressed by contralateral white noise, abnormal ABR, no acoustic reflex and--SP/AP > 0.44. CONCLUSIONS: The results suggest that cochlear outer hair cells are normal in function with dysfunction of the cochlear nuclei. The main lesion of the disease are neural pathways related to Acoustic reflex and medial olivo-cochlear efferent nervous system. The cochlear afferent nerve may also be involved. PMID- 12768757 TI - [Cholesteatoma of the cerebellopontine angle presented as trigeminal neuralgia]. AB - OBJECTIVE: To explore the effective methods of diagnosis and treatment for cholesteatoma of the cerebellopontine angle presented as trigeminal neuralgia. METHODS: The Clinical data of 13 patients were studied retrospectively. RESULTS: Only 1 case of Cholesteatoma in cerebellopontine and suprasellar cistern was detected with cranial CT scans among 13 patients. Prolonged latency of Wave V and interpeak peak latencies of I-V Wave were recorded in 1 case on auditory brainstem response(ABR). But 9 patients had abnormality in ENG (2 paralysis, 2 paresis). During the operations, we found that cholesteatoma tissue or its capsule wrapped the roots of trigeminal nerves. The cholesteatoma were removed through post-sigmoid sinus approach without partial amputation of trigeminal posterior root. All the patients were clinically cured by extraction of cholesteatoma with following up period of 3 years (10 cases) and half of a year(3 cases). Cholesteatoma extended above tentorium was completely resected with post sigmoid sinus approach in one case. CONCLUSIONS: ENG is more sensitive than cranial CT and ABR for diagnosis of cholesteatoma in the cerebellopontine angle. The post-sigmoid sinus approach is the best operative approach. It is unnecessary to have the partial amputation of trigeminal posterior root if cholesteatoma can be completely removed. When cholesteatoma extends above tentorium, it can be removed simultaneously by post-sigmoid sinus approach. PMID- 12768758 TI - [Treatment of traumatic cerebrospinal rhinorrhea in frontal sinus]. AB - OBJECTIVE: To evaluate the indication, operative technique and advantages of the treatment of traumatic cerebrospinal rhinorrhea in frontal sinus by intranasal endoscopic surgery. METHOD: Treatment of 6 cases of traumatic cerebrospinal rhinorrhea in frontal sinus by intranasal endoscopic surgery was reported. We adopt a procedure to open and enlarge the nasofrontal duct under direct vision by intranasal endoscopic surgery firstly and find the fistula, then repair it with its own smashed muscle, and support it with muscular membrane and gelfoam, iodoform Sponges. RESULTS: Four cases were repaired successively in one procedure with intranasal endoscopic surgery, 2 cases were solved by combined external and intranasal procedure. CONCLUSIONS: The advantages of treatment of traumatic cerebrospinal rhinorrhea in frontal sinus by intranasal endoscopic surgery included easily operation, easily confirmation of fistula and high success rate, combination of frontal sinus incision may deal with its demerit to the un-reach able site sometimes. PMID- 12768759 TI - [Determination of the immune function of the erythrocyte and leucocyte and the activity of IL-4 and IFN-gamma in patients with allergic rhinitis]. AB - OBJECTIVE: To investigate the immune function of erythrocyte (RBC) and leucocyte (WBC), and variety of the interleukin-4 (IL-4) and interferon-gamma (IFN-gamma) among patients with allergic rhinitis. METHODS: IL-4 and IFN-gamma secreted by peripheral blood mononuclear cells (PBMC) were measured and compared with immune adhesive and BA-ELISA method. RESULTS: Results showed the RBC and WBC immune adhesive function and IFN-gamma were significantly decreased than of the control, and IL-4 activity were significantly higher than the controls (P < 0.01) in patients with perennial allergic rhinitis. CONCLUSION: These results suggest that RBC and WBC the immune function and IL-4 and IFN-gamma may play an important role in allergic rhinitis. PMID- 12768760 TI - [Study on the relationship between the abnormal expression of c-myc and multidrug resistance in KB cell lines]. AB - OBJECTIVE: To investigate the relationship between expression of c-myc and the regulation of mdr1, and find out the best way for gene therapy of the proliferative diseases with c-myc overexpression and reverse the multidrug resistance phenotype. METHODS: The changes of c-Myc protein and P-gp in the KB cell lines were detected by using immunohistochemistry and flow cytometry methods before and after treatment with anti-mdr1- ribozyme. RESULTS: All KB cell lines could express c-Myc protein. The expression of c-myc in multidrug resistant cell KBv displayed higher than that in sensitive cell KB. However, after reversing the multidrug resistance phenotype by anti-mdr1-ribozyme, the level of c-Myc protein expression was lower in KBv/5mR3 and KB/5mR3. The level of P-gp expression was lower in KBv/5mR3 than that in KBv but no difference in KB/5mR3 and KB. CONCLUSION: There are a close relationship between Myc protein and MDR phenotype. The c-myc involves in regulating expression of mdr1. Myc protein can be used as a new assistant implement to monitor the formation of MDR. PMID- 12768762 TI - [The status of protein P16 and P53 in the surgical margins of laryngeal carcinoma and the association with local recurrence]. AB - OBJECTIVE: To assess the status of P53 and P16 of surgical margins in laryngeal carcinoma and the association with the local recurrence following surgical treatment METHODS: The status of P53 and P16 of surgical margins in laryngeal carcinoma was studied using immunohistochemistry (IHC) staining method in 100 cases of laryngeal carcinoma. Fifty-two of them had recurrent tumors while 48 cases were free of local recurrence after 5-year follow up. The original tumor sites and T stages, which bear important influence over the risk of local recurrence, were strictly matched between the two groups, and the ages and preoperative doses of radiation were also considered. RESULTS: The percentage of local recurrence was 85% in these patients who showed positive staining for P53 and simultaneous negative staining for P16 in both the cancer tissues and the surgical margins. No association was found between the expression of P53 and P16 and the clinical types or T stages. CONCLUSION: The status of P53 and P16 in combination demonstrated by IHC technique in surgical margins of laryngeal carcinoma may predict the possibility of local recurrence. PMID- 12768761 TI - [Basement membrane in squamous cell carcinomas of the larynx: an immunohistochemical study]. AB - OBJECTIVE: The study was to assess the distribution of basement membrane(BM) in laryngeal squamous cell carcinomas(LSCC). Correlation of BM and clinical parameters (TNM stage, histological grading, invasion mode, lymph node metastasis) was also examined. METHODS: The expression of BM around tumor cell was determined in 40 cases of LSCC by using monoclonal antibody against human type IV collagen. An intact continuous BM was found in 17 cases (42.5%), while partial or widespread loss of the BM was detected in the other 23 cases (57.5%). RESULTS: In cases with poor histological differentiation, the defect of BM was more severe than that in cases with high or middle histological differentiation (P < 0.05). Moreover, diffuse invasion carcinomas revealed lower type IV collagen expression comparing with cases with better tumor-host border (P < 0.05). There was a higher risk of regional lymph node metastasis among cases with poor BM expression (P < 0.01), but there was no association of clinical stage with BM defect (P > 0.05). CONCLUSION: These observations indicated that testing the distribution of BM seems to be useful to evaluate the histological grading of malignancy of laryngeal carcinoma and be helpful to prognosticate the frequency of regional lymph node metastasis. PMID- 12768763 TI - [Studies on c-myc gene expression and p16 gene inactivation in nasopharyngeal carcinoma]. AB - OBJECTIVE: The alterations of c-myc and p16 genes and their relationship with the development and progression of nasopharyngeal carcinoma were studied. METHODS: Sixty-nine biopsies of nasopharyngeal carcinoma were examined for the homozygous deletion methylation and reduced expression of p16, and the overexpression of myc family oncogenes using multiple PCR, restriction endonuclease coupled PCR, reverse transcriptase PCR and immunohistochemistry. RESULTS: The homozygous deletion and methylation of p16 gene were found in 7 and 11 cases respectively. The total inactivation rate was 26.1% (18/69). The negative expression of p16, as shown by immunohistochemical examination was found in 42 out of 69 cases (60.9%). The overexpression of myc family oncogenes was found in 51 cases (73.9%). CONCLUSION: It suggests that the inactivation of anti-oncogene p16 and the activation of myc family oncogene may play an important role in the development of nasopharyngeal carcinoma. PMID- 12768764 TI - [Clinical significance of elevated serum sICAM-1 in patients with nasopharyngeal carcinoma]. AB - OBJECTIVE: To study the expression of serum sICAM-1(soluble intercellular adhesion molecule-1) and tissue ICAM-1 in nasopharyngeal carcinoma and its clinical significance. METHODS: Expression of ICAM-1 in frozen sections of 20 normal nasopharyngeal mucosa and 54 nasopharyngeal carcinoma samples were studied with immunohistochemical technique, and levels of serum sICAM-1 in 54 patients with nasopharyngeal carcinoma (NPC group) and 32 healthy adults (normal group) by ELISA. RESULTS: Positive expression (45/54) of tissue ICAM-1 and levels (2,044.48 +/- 640.05) microgram/L of serum sICAM-1 in NPC group were significantly higher than that of 2/20 and (313.70 +/- 114.98) microgram/L in normal group (P < 0.01, respectively), and there was a positive relationship between the expression of tissue ICAM-1 and levels of serum sICAM-1 in nasopharyngeal carcinoma(P < 0.05). The levels (2,177.31 +/- 642.13) microgram/L of serum sICAM-1 in patients with III-IV stages were higher than those (1,847.25 +/- 308.15) microgram/L in I-II stages (P < 0.05), and those (1,678.89 +/- 685.01) microgram/L in patients with no lymph node metastasis lower than that (2,134.34 +/- 609.60) microgram/L in lymph node metastasis (P < 0.05). The levels of serum sICAM-1 decreased significantly from (2,211.42 +/- 717.59) microgram/L to (926.55 +/- 207.72) microgram/L in disappearance of tumor and metastatic lymph node after radiotherapy (P < 0.01). CONCLUSION: sICAM-1 might be involved in the cellular immunopathogenesis of nasopharyngeal carcinoma, and the measurement of the serum sICAM-1 may reflect ability of neoplasm cell in freedom from host immunological surveillance. Dynamic measurement of serum sICAM-1 may be a useful objective index to evaluate NPC prognosis and curative effects. PMID- 12768765 TI - [Infiltration of dendritic cells into cervical lymph nodes in laryngeal carcinoma]. AB - OBJECTIVE: Dendritic cells(DC) form a system of highly efficient antigen presenting cells. An immunohistochemical study was performed to prove their role in immune responses against cancer. METHODS: Paraffin blocks were prepared for staining with antibody against S-100 proteins in 157 lymph nodes obtained from elective cervical lymphadenectomy in 47 cases of laryngeal squamous cell carcinomas. RESULTS: The extent of infiltration by DC into the regional lymph nodes was evaluated in two groups according to negative and positive lymph node metastasis. Densities of DC ranged from 23.0 to 111.9, with an average of 57.2 +/ 16.0, in the negative metastasis nodes; and from 17.5 to 63.6, with an average of 38.3 +/- 14.9, in the positive metastasis nodes. The test revealed that the differences in population density between the two groups were significant (P < 0.001). Quantitative analysis showed that patients who survived longer than 5 years had significant higher numbers of DC in the lymph nodes than those who lived less than 5 years (P < 0.001). Densities of DC ranged from 32.0 to 111.9, with an average of 61.6 +/- 19.6, in the nodes of 20 cases of patients who survived longer than 5 years and from 17.5 to 65.8, with an average of 37.9 +/- 13.7, in the nodes of 27 cases of patients who survived less than 5 years. CONCLUSION: DC may be more directly involved in the host immune reaction against tumor by means of the most important antigen-presenting cells. DC in the cervical lymph nodes are important for establishing immunologic defense mechanisms in cases of laryngeal carcinomas, and the densities of DC can serve as a useful indicator in assessing prognosis of laryngeal carcinomas. PMID- 12768767 TI - [Surgery for tumors of the root of neck]. AB - OBJECTIVE: To investigate the clinical value of the surgery for tumors located in the root of the neck. METHODS: A retrospective study was made on 15 cases with tumors in the root of the neck. Seven patients had benign and 8 had malignant tumors. In the malignant group, 4 staged III and 4 staged IV. Patients with benign disease were followed up for more than 3 years, and patients with malignant tumors were followed up from 1 to 5 years (median 2 years). RESULTS: Resection via "L" incision in the root of the neck was employed in 13 cases (7 benign and 6 malignant, clavicle splitting in 1 case) and sternotomy in 2 cases. Only one case in the benign group had complication (chylous leakage). In contrast, 7 complications (2 subclavian vein tearing, 2 brachial plexus injury, 2 chylous leakage and 1 pneumothorax) occurred in 6 patients in the malignant group. The incidence of complication was significantly different between the two groups (P < 0.05). We observed no recurrence in both groups during the period of follow-up. CONCLUSIONS: This study showed that surgery of tumors located in the root of the neck is effective, although the incidence of surgical complication is high for the malignant ones. PMID- 12768766 TI - [Study of mechanisms of lightening aspiration and increasing decannulation rate in anastomosis of cricoid cartilage and base of tongue (epiglottis)]. AB - OBJECTIVE: To study laryngopharyngeal anastomosis and fixing methods of super cricoid laryngectomy with reconstruction of functions in lightening aspiration and increasing decannulation rate. METHODS: Recovering conditions of laryngeal functions in 66 patients who underwent supracricoid laryngectomy and anastomosis of cricoid cartilage and base of tongue (epiglottis) in recent eight years were summarized. Some relevant caliber distances in 21 residual larynges were measured. RESULTS: All cases restored their phonation. In 36 cases that underwent anastomosis of cricoid cartilage and base of tongue, 15 cases without aspiration, 18 with mild aspiration, 3 moderates. Decannulation rate is 94.4%. In 30 cases who underwent anastomosis of cricoid cartilage and base of tongue, 10 with mild aspiration, 17 moderate, 3 serious. All cases overcame aspiration within 3 weeks. 7 cases were cannulated. 3, 5, 10 year survival rates were 80.3%, 74.4%, and 3/7. The longitudinal and transverse calibers of epiglottis were 1.5-2.0 times longer than that of the entrance of cricoid cartilage. After anastomosis of cricoid cartilage and base of tongue (epiglottis), the epiglottis can exactly cover the entrance of cricoid cartilage to prevent aspiration fully and increase decannulation rate. Previously the cricoid cartilage was anatomized and fixed under the hyoid bone. Because some spaces exist between cricoid cartilage, base of tongue and epiglottis, aspiration is likely to occur. That hyoid bone covers the entrance of cricoid cartilage will bring constriction of the new laryngeal orifice and make decannulation difficult. Hyoidectomy and anastomosis of cricoid cartilage and base of tongue (epiglottis) overcame the two shortcomings and had good effects. CONCLUSION: Cricoid-hyoid-anastomosis was the main reason of severe aspiration and low decannulation rate. The ideal methods to lighten aspiration and increase decannulation rate are hyoidectomy and anastomosis of cricoid cartilage and base of tongue (epiglottis). PMID- 12768768 TI - [Treatment of cancer of the nasal cavity in 128 cases]. AB - OBJECTIVE: To investigate the credibility of combined radiotherapy and surgery for cancer of the nasal cavity and their prognostic factors. METHODS: One hundred and twenty-eight patients were treated from Mar. 1977 to Aug. 1993 receiving either radiotherapy alone (R group, 89), radiotherapy plus surgery (R + S group, 21), surgery plus radiotherapy (S + R group, 13), or surgery alone (S group, 5). RESULTS: The overall 5-year survival rate was 42.2%. The 5-year survival rates of R, R + S and S + R groups were 33.7%, 57.1% and 61.5%, respectively. The 5-year survival rate of combined therapy for squamous cell carcinoma and low differentiated cancer were higher than that of radiotherapy alone, and the effect of radiotherapy alone for undifferentiated carcinoma was better. In early stage, the 5-year survival rate of the radiotherapy alone and combined therapy did not differ significantly. For III and IV stages, the 5-year survival rate of R + S combined therapy was higher than those with radiotherapy alone (P < 0.01). CONCLUSIONS: For early lesions(Stage I and II), R alone or R + S give better results. For advanced lesions(Stage III and IV), combined radiotherapy and surgery is preferred. Radiotherapy alone is indicated for poorly differentiated cancers. Combined radiotherapy plus surgery is preferred for squamous cell carcinoma. PMID- 12768769 TI - [The clinical significance of poststyloid space extension in nasopharyngeal carcinoma]. AB - OBJECTIVE: To explore the clinical significance of poststyloid space extension in nasopharyngeal carcinoma. METHODS: One hundred and ninety-eight patients with newly treated nasopharyngeal carcinoma (stage I-IV a) were analyzed. These patients were divided into two groups depending on whether the poststyloid space was extended or normal. The 5-year survival rate, relapse and distant metastasis after the treatment were estimated using the Kaplan-Meier, Log-Rank methods. RESULTS: The rates of patients with oropharyngeal extension for these two groups were 72.1% and 16.8% respectively (P < 0.001). The rates of the cases with involvement of the 9, 10, 11, 12th cranial nerves were 20.9% and 6.2% respectively (P < 0.01). The rates of metastasis to cervical lymph node were 69.8% and 35.4% respectively (P < 0.01). The 5-year survival rates were 46.76% and 68.88% respectively (P = 0.0027). The local control rates were 63.7%, 78.7% respectively (P = 0.0862). The distant metastasis free survival rates were 71.7% and 84.2% respectively (P = 0.0468). These results showed that the survival rate, the local control rate were reduced and the distant metastasis was increased for patients with poststyloid space extension. CONCLUSIONS: The poststyloid space extension was an important prognostic factor with poorer treatment outcome. It is reasonable to classify such condition as stage T3 of nasopharyngeal carcinoma. PMID- 12768770 TI - [Lateral neck dissection of hypopharyngeal cancer with clinically regional metastasis]. AB - OBJECTIVE: To assess whether lateral neck dissection can control the hypopharyngeal cancer with clinically regional metastasis. METHODS: A retrospective review of medical chart from 1975 to 1992 was performed. Ninety three hypopharyngeal squamous cell carcinoma patients who had performed RND were included. The distribution of metastatic neck lymph node was analyzed. RESULTS: Submandibular triangle lymph node metastasis was occult in only 3 patients. Histological lymph node metastasis to the posterior triangle was found in 5.9% of patients with N0, 7.0% with N1, 37.5% with N2a and 36% with N2b-N3. Histological neck lymph node metastasis to the posterior triangle was found in 4.0% of patients without inferior jugular lymph node metastasis and 34.1% of patients with inferior jugular lymph node metastasis. CONCLUSIONS: Lateral neck dissection was recommended to treat hypopharyngeal cancers of N0 and N1. N2 and N3 should be treated with neck dissection including II-V group lymph nodes. After lateral neck dissection, frozen section of the inferior jugular lymph node should be performed. If the result of the frozen section is positive, V group dissection should be performed. PMID- 12768771 TI - [Supracricoid partial laryngectomy in laryngeal cancer]. AB - OBJECTIVE: To assess the effectiveness of supracricoid partial laryngectomy in the treatment of laryngeal cancer and the reconstructed laryngeal functions. METHODS: This study involved 29 patients operated on from 1978 to 1998 using this surgical procedure. In these cases, 18 were male and 11 were female with a mean age of 55.4 years (range, 38 to 70 years). The disease was classified according to the 1987 UICC system. There were 10 supraglottic cancers (3 T2N0, 1 T2N1, 3 T3N0, 2 T3N1, 1 T3N2), and 19 glottic cancers (12T2N0, 1T2N1, 4T3N0, 2T3N1). Two subtypes of supracricoid partial laryngectomy were performed, one with epiglottis preserved and reconstructed with cricohyoidepiglottopexy (CHEP), the other involved epiglottic resection and reconstructed with cricohyoidopexy (CHP). RESULTS: The voice was good in all cases. Twenty-seven cases were decannulated (93.1%). All cases had overcome the aspiration by practice. The overall 3-year and 5-year survival rates were 89.7% and 75.9%. CONCLUSION: Supracricoid partial laryngectomy was appropriate for selected supraglottic and glottic cancers. PMID- 12768772 TI - [Supracricoid subtotal laryngectomy for the treatment of advanced laryngeal cancers]. AB - OBJECTIVE: This procedure was designed to eradicate the advanced laryngeal carcinomas while preserving the laryngeal physiologic function. The indications and contraindications for the operation were also discussed and evaluated in this paper. METHODS: Twenty-one patients with advanced laryngeal cancers underwent this procedure during the period from 1988 through 1996 in our department. All of the patients were not amenable to conventional partial horizontal and vertical laryngectomies. In those patients, there were 9 supraglottic cancers, 10 glottic cancers and 2 transglottic cancers. Among them, 16 cases had stage T2 lesions and 5 cases had stage T3 lesions. In this procedure, the hyoid and thyroid cartilages were excised in toto, along with both true and false vocal cords, paraglottic space as well as the partial or total epiglottis and the whole preepiglottic space. The cricoid cartilage and at least one arytenoid cartilage were spared. The new larynx was reconstructed by suturing the cricoid and/or epiglottis to the base of the tongue. RESULTS: Of 21 patients, 95% were able to eat normally without aspiration, 90% were decannulated with near normal speech function. In two patients, the tracheotomy tube was reserved, one developed a stenosis, and another had recurrent aspiration pneumonia. No patients experienced local recurrence. Nineteen patients were followed up for at least 3 years, 13 patients achieved 5 years tumor-free survival. CONCLUSION: The term supracricoid subtotal laryngectomy best describes the surgical approach and the reconstruction model of the new larynx. As a useful alternative to total laryngectomy in selected advanced laryngeal cancers, this procedure may greatly improve the quality of life and prolong the life without recurrence. PMID- 12768773 TI - [Detection of bcl-2/JH gene rearrangement by semi-nested polymerase chain reaction from fresh tumor samples in patients with laryngeal squamous cell carcinomas]. AB - OBJECTIVE: To evaluate the clinical implications of bcl-2/JH gene rearrangement in laryngeal squamous cell carcinomas (LSCC). METHODS: Bcl-2/JH gene rearrangement analysis in fresh tumor samples was performed in 60 patients with LSCC by semi-nested polymerase chain reaction(PCR). RESULTS: The results showed that bcl-2/JH gene rearrangement was found in 37 out of 60 patients. The breakpoint was located within the major breakpoint region(mbr) in 33 of the 60 patients and the remaining patients had bcl-2 translocation within the minor cluster region(mcr). The results of the study showed that the rearrangement rate of bcl-2 gene was not related to the grade of differentiation, clinical stage, and neck lymph-node metastasis (P > 0.05), and it was related to heavy smoking (P < 0.05). CONCLUSION: Detection of bcl-2/JH gene rearrangement could reveal that bcl-2/JH fusion gene in LSCC is an important molecular biological marker and has a significant role in occurrence and development of LSCC. PMID- 12768774 TI - [Mutations in the connexin 26 gene in patients with nonsyndromic hearing impairment]. AB - OBJECTIVE: To determine the prevalence and characteristics of deafness-causing mutations in Connexin 26(Cx26, GJB2) gene in Chinese with nonsyndromic hearing impairment(NSHI). METHODS: Study subjects are all Chinese including 16 infants with sporadic congenital deaf-mutism, 39 patients with autosomal recessive hereditary hearing loss, 30 patients with autosomal dominant hereditary hearing loss and 100 normal adults. The subjects were screened for base variations by single-strand conformational polymorphism (SSCP) analysis of the amplified products of polymerase chain reaction (PCR). Those who were found have abnormal conformational band were sequenced. RESULTS: Five kinds of polymorphism were found in 15 cases of controls and six kinds of polymorphism in 10 patients. No mutation was found in Cx26 gene in Chinese with autosomal recessive NSHI. Heterozygous deletion AT at position 299-300 of Cx26 cDNA, which results in premature chain termination, was found in a pedigree with autosomal dominant hereditary nonsyndromic hearing loss. CONCLUSION: The prevalence of deafness causing mutations in Cx26 gene in Chinese with autosomal recessive NSHI maybe is lower than that of other ethnic groups. Heterozygous deletion AT at position 299 300 of Cx26 cDNA can lead to autosomal dominant hereditary hearing loss (DFNA3). PMID- 12768775 TI - [Thiol reagent thimerosal-induced Ca2+ mobilization in isolated guinea pig cochlear outer hair cells]. AB - OBJECTIVE: To understand mechanism of cochlear outer hair cells (OHCs) intracellular Ca2+ mobilization further. METHODS: Intracellular calcium of isolated guinea pig was investigated using thimerosal, a--SH group oxidizing agent, and fura-2 fluorescence ratio imaging microscopy. RESULTS: In the presence of thimerosal, intracellular Ca2+ concentrations ([Ca2+]i) of OHCs were elevated in a dose-dependent manner. Even in Ca(2+)-free medium, Ca2+ response was still induced. The effects of thimerosal on [Ca2+]i were completely blocked and reversed by (DTT). Neither 1-100 mumol/L ryanodine nor 5-20 mmol/L caffeine altered the effects of thimerosal. Pretreatment with pertussis toxin (PTX) for 30 min did not affect the thimerosal-induced increase in [Ca2+]i The increase in [Ca2+]i when Ca2+ was added during thimerosal application in Ca(2+)-free medium was almost completely blocked by 500 mol/L LaCl3, while nifedipine did not inhibit further increase in [Ca2+]i caused by thimerosal. CONCLUSION: Oxidation of the -SH group of the OHC membrane can induce a Ca2+ release from intracellular Ca2+ stores, which are ryanodine- and caffeine-insensitive, and Ca2+ influx through non-specific Ca2+ channels, but not the nifedipine-sensitive Ca2+ Channels. The possible oxidation of--SH group gated Ca2+ channels in OHCs are worthy of further study. PMID- 12768776 TI - [An observation on long-term influence of middle ear bacterial infection on inner ear function and systemic immune reaction]. AB - OBJECTIVE: To understand whether long-term inner ear heat shock response related to heat shock protein(HSP70) caused by middle ear bacterial infection and the potential influence on inner ear function. METHODS: Sixty BALB/c mice were randomly classified into 6 groups including Klebsiella pneumoniae (KP), Staphylococcus aureus, Bacillus pyocyaneus, Bacillus coli, Bacillus proteus and physiological saline control groups. On 135 days after injection, distortion product otoacoustic emissions(DPOAE) was tested and all the samples were collected, which were examined with light and electronic microscopes. HSP70 related molecule expression in inner ear, nuclear factor (NF) kappa Bp65 characterization in mononuclear cell, anti-KP antibody and anti-membranous labyrinth proteins (MLP) were examined. RESULTS: No nuclear transfer of NF kappa Bp65 was observed in any animal. Anti-KP antibody was detected in 30% (3/10) of Staphylococcus aureus group, 29% (2/7) of KP group, 33% (3/9) of Bacillus pyocyaneus group and 10% (1/10) of control group. Anti-MLP antibody was created in 20% (2/10) of Staphylococcus aureus group, 20% (2/10) of KP group, one each in Bacillus pyocyaneus group and control group respectively. Double positive antibody against KP and MLP were found in Staphylococcus aureus group and KP group. When analyzed with Western blot, all the positive bands were small molecules including strongest 26,000-30,000, medium degree 38,000-41,000 and weak 46,000-50,000 except for 68,000 in one case. There was only one significant DPOAE amplification decrease at 1,625 Hz (2f1-f2) in left ear of Bacillus pyocyaneus group and right ear of Bacillus proteus group. No abnormal phenomenon was found in inner ear both under light microscope and electronic microscope. No significant expression of HSP70 was observed in inner ears. CONCLUSION: No long term heat shock response related to HSP70 existed in the inner ear and the immune inner ear damage may be caused by multiplefactors. PMID- 12768777 TI - [Outward potassium currents in isolated Deiters' cells of guinea pig cochlea]. AB - OBJECTIVE: To study potassium currents in isolated Deiters' cells of guinea pig cochlea. METHODS: Potassium currents in Deiters' cells in normal external solution, in external solutions with different K+ concentrations effecting on reversal potentials and outward potassium currents, 4-aminopyridine (4-AP) and tetraethylammonium (TEA) blocking components of outward potassium currents and kinetics of activation and inactivation of potassium ion channels were studied using the whole cell variant of the patch clamp technique. RESULTS: Isolated Deiters' cells could possess voltage dependent, outwardly rectifying ion channel which was K+ selective. Existence of two separate types of K+ channels or their closed and open states was suggested by effects of 4-AP and TEA. Activation and inactivation of ion channels were described by Boltzmman equation. Voltage dependent inward Ca2+ current could not be detected. CONCLUSION: Outwardly rectifying potassium in Deiters' cells could buffer extracellular K+ in the small space between Deiters' cells and outer hair cells or neural fibers and alter the stiffness of Deiters' cells, which as a result might influence cochlear mechanics. PMID- 12768778 TI - [Three-dimensional reconstruction of internal auditory meatus and anatomical study of the inner structures]. AB - OBJECTIVE: To provide anatomical basis for surgical operation in internal auditory meatus (IAM). METHODS: Structures of cerebellopontine angle (CPA) and IAM in 25 temporal bone specimens (50 sides) fixed in formalin were measured. Three-dimensional reconstruction (3DR) of IAM in 13 normal adults (26 sides) was conducted using spiral CT scanning. RESULTS: IAM took a conic shape with larger inside and smaller outside. The average width of IAM was (4.16 +/- 1.23) mm(2.87 6.83 mm) in horizontal dimension, the average height was (3.14 +/- 1.01) mm(2.23 4.45 mm) in vertical dimension, the average length was (8.67 +/- 2.31) mm (6.77 11.22 mm), the average volume was (98.23 +/- 16.56) mm3(67.44-133.21 mm3). There were 1-4 internal auditory arteries branched from the anterior-inferior cerebellar artery, which formed a vascular ansa near the opening of IAM. The average length of the facial nerve in IAM was (0.98 +/- 0.03) mm (0.89-1.07 mm), the average diameter of vestibulocochlear nerve was (1.67 +/- 0.05) mm (1.42-1.97 mm), the average diameter of the intermediate nerve (0.23 +/- 0.02) mm (0.19-0.26 mm). CONCLUSION: It is necessary to pay attention to types of vascular ansa and relative nerve location in IAM during operation. PMID- 12768779 TI - [Neurofibromatosis type 2]. AB - OBJECTIVE: To investigate the clinical characteristics and management strategies of bilateral acoustic neuromas. METHODS: The data of 7 patients with bilateral acoustic neuromas collected between 1990 to 1998 were retrospectively analyzed. RESULTS: Altogether 122 patients with acoustic tumors were treated from 1990 to 1998, in which 7 cases (5.8%, 6 male and 1 female) had bilateral acoustic neuromas. The age at onset of symptoms ranged from 13 to 60 years (average 29.1 years). Progressive hearing loss and tinnitus were the initial symptoms in 4 cases. Either strabismus, ptosis, headache or dysequilibrium was presented in 4 cases. Six cases complicated tumors in the central nervous system and/or other sites. Five cases had cafe au lait spots. One case had posterior subcapsular lenticular opacity. Four cases fell into severe (Wishart) type and 2 into mild (Gardner) type. The tumors were unilaterally removed in 4 patients through the retrolabyrinthine approach (1 case) or the retrosigmoid approach (3 cases). In these 4 patients, one died of central respiratory failure after the operation; two had contralateral tumor removal through retrosigmoid approach 3 weeks after the first surgery. One of the patients died of encephaledema after the surgery. No hearing impairment and facial nerve paralysis occurred in one case operated on through the retrolabyrinthine approach, whereas in those through retrosigmoid approach, 4 ears had hearing loss and 3 sides had facial nerve paralysis. CONCLUSION: The clinical characteristics and treatment strategies for bilateral acoustic neuromas are different from those of unilateral acoustic neuroma. Individualization of management is a prerequisite for the success of the treatment. To avoid injury to the VII and VIII cranial nerves, monitoring the nerve functions during the surgery is important. PMID- 12768781 TI - [Value of preservation of uvula in uvulopalatopharyngoplasty]. AB - OBJECTIVE: To explore the effect of the modified Uvulopalatopharyngoplasty (UPPP) where uvula is preserved in the operation, and whether the postoperative complications are diminished. METHODS: Thirty consecutive patients with the obstructive sleep apnea syndrome (29 men, 1 woman) were analyzed by questionnaire, polysomnography and oral cavity measurement before and 6 months after uvulopalatopharyngoplasty to assess the surgical outcome and value of preservation of uvula in the operation. RESULTS: Eighty-seven percent of patients reported improvement of snoring and daytime sleepiness. Fifty-three percent of patients showed a decrease of at least 50% in the apnea and hypopnea indices. Among the mild and moderate obstructive sleep apnea (AHI < 50), 73.3% of them showed a decrease of at least 50% in the AHI. No velopalatal insufficiency occurred. The result of oral cavity measurement showed that the uvula, which was preserved completely during the operation, began to contract 2 weeks after the operation. Three to six month after operation, uvula contracted to the normal length and the anatomical shape of the pharyngeal cavity became normal. CONCLUSIONS: The results of the uvula preserving UPPP indicate that this approach not only improves the effect of the UPPP, but also avoids the postoperative complications. The uvula that preserved completely during the operation will contract to the normal length progressively. PMID- 12768780 TI - [Value of condensation and rarefaction click evoked action potential latency difference in the diagnosis of Meniere's disease]. AB - OBJECTIVE: To study the value of condensation and rarefaction clicks evoked action potential (AP) latency difference (LD) in diagnosis of Meniere's disease. METHODS: AP was recorded with ECochG in controls (50 ears) and patients with Meniere's disease(90 ears) and sensorineural hearing loss(SNHL) of other origins(60 ears). LD was calculated and analyzed. RESULTS: LD in patients with Meniere's disease was (0.30 +/- 0.15) ms, which was significantly larger than that of controls(0.18 +/- 0.07) ms and of patients with SNHL of other origins(0.20 +/- 0.10) ms (P < 0.01). In the group of Meniere's disease, LD in patients with the mild and moderate hearing impairment was larger than those with severe hearing loss(P < 0.01) and LD in patients with low tone or high tone auditory sensation curve was larger than those with flat auditory sensation curve(P < 0.01). Positive rate was 4/60(6.7%) in other SNHL patients and 58/90(64.0%) in Meniere's disease group respectively. CONCLUSION: The increase in condensation and rarefaction click evoked AP latency difference can be an objective parameter in diagnosis of Meniere's disease. PMID- 12768782 TI - [The experience with endoscopic natural ostiotomy]. AB - OBJECTIVE: To improve the operative technique of endoscopic middle metal antrostomy. METHODS: One hundred and thirty-five patients with chronic sinusitis were divided into two groups(first stage and second stage groups). All of them had been followed up for more than 6 months. RESULTS: Middle metal antrostomy was performed for 79 patients (158 sides) in the first group. The diameter of middle meatal window ranged from 1.0 cm (anterior-posterior diameter) to 1.5 cm(superior inferior diameter). Improved middle meatal antrostomy was performed for 56 patients (112 sides) in the second group. The anterior-posterior diameter of the middle meatal window was over 1.0 cm, and superior-inferior diameter was over 1.5 cm. In the first group, the patency rate in the first stage was 78.48% (124/158), and satisfactory patency rate was 43.0% (68/158). The patency rate was 85.4% (135/158), and satisfactory patency rate was 45.0%(71/158). In the second group, the patency rate was 92.9%(104/112) in the first stage, and 94.6%(106/112) in the second stage. The satisfactory patency rate in both stages was 79.5%(89/112). There was significant difference between two groups(P < 0.05). CONCLUSION: The patency rate and satisfactory patency rate of middle meatal antrostomy are increased by improved technique. PMID- 12768783 TI - [Complications and its prevention in 227 cases of childhood tracheotomy]. AB - OBJECTIVE: To explore the correlation between the occurrence and the prevention of complications in childhood tracheotomy. METHODS: A total of 227 cases of childhood tracheotomy were analyzed based on clinical data from March 1974 to July 1999. RESULTS: Of 227 cases, 189 were decannulated successfully. Twenty three cases discharged with tracheotomy tube. Fifteen cases died of primary disease. Fourteen of 227 cases had complications. Among them, 2 had intraoperative suffocation, 1 had trauma of esophagus, 5 complicated subcutaneous emphysema in cervical region and 1 had mediastinal emphysema. Five of 14 cases had postoperative drop out of the tube, 2 of them died of suffocation. CONCLUSION: The occurrence of complications is related to age, original disease, operative method and nursing. PMID- 12768784 TI - [A long-term observation of 48 cases of tracheoesophageal shunt phonation by the anastomosis of the membranous portion of the tracheal section with the anterior wall of esophagus after total laryngectomy]. AB - OBJECTIVE: To study the long-term efficacy of tracheoesophageal (TE) shunt phonation by the anastomosis of the membranous portion of the tracheal section with the anterior wall of esophagus after total laryngectomy. METHODS: A questionnaire was designed to follow 48 patients with rehabilitated speech and swallow functions by the above approach. The qualities of phonation and speech, and the degree of aspiration evaluated, together with the survival rate and complications statistically analyzed. RESULTS: The speech level of 35 laryngectomees was similar to the normal laryngeal speech level. Thirty-five cases had longer maximal phonation time and hearing distance and higher speech intelligibility. Other 5 laryngectomees had less effective phonation, but higher speech intelligibility. The total effective rate was 83.3% (40/48). The speech fluency in 40 laryngectomees was not as good as that in the normal people. Forty percent laryngectomees(16/40) had slight liquid food aspiration which did not influence normal eating. Eight patients(8/48) failed to speak and no liquid aspiration occurred after the operation. The effective rate was affected obviously by neck infection and pharyngeal fistula formation. The survival rate was similar to those with single total laryngectomy. CONCLUSIONS: The TE shunt phonation had the advantages of simple, one-stage operation and high success rate. Some laryngectomees had slight liquid food aspiration, but would not influence normal eating. So this TE shunt phonation operation may be useful during laryngeal surgery. PMID- 12768786 TI - [Quality of life following laryngectomy]. AB - OBJECTIVE: To assess the quality of life and relevant factors following laryngectomy. METHODS: Quality of life was evaluated in 42 laryngeal cancer and hypopharyngeal cancer patients following laryngectomy and in 38 controls. The assessment included General Quality of Life Inventory, University of Washington Quality of Life Head and Neck Questionnaire and a study specific questionnaire. RESULTS: The laryngectomized patients reported significant lower scores of quality of life than the controls in many aspects of physical health, psychological health and social function dimensions. For patients, the most important dimension affecting quality of life was psychological condition, and the others were physical function, social function, and economic condition in turn; while for the controls, they were of opposite order. The significant factors affecting quality of life in patients following total laryngectomy were their concern about prognosis, disfigurement, decannulation, employment, speech and breathing conditions. The most important factor affecting psychological condition was physical function, while psychological condition was the most important factor affecting social function. Items ranked in the top five included those related to physical function, economic condition, psychological condition, social situation and social communication. CONCLUSION: More attention should be paid to research about quality of life. The quality of life of patients undergone laryngectomy is affected by many factors. Through the integration of psychological, clinical and social interventions, the quality of life of patients may have an overall improvement. PMID- 12768785 TI - [Mechanism of esophageal speech after total laryngectomy]. AB - OBJECTIVE: To evaluate the characteristics of esophageal speech after total laryngectomy. METHODS: Esophageal speech evaluation of 40 cases of esophageal phonation included acoustic parameters, intraesophageal pressures during phonation, speech intelligibility, fluency, communication, respiratory sound and cognate distinctions between voiced and voiceless sounds. RESULTS: In 8 poor speakers, the neoglottis was spasmodic and difficult to vibrate during phonation. In 32 good speakers, the neoglottis was relaxed and easy to vibrate during phonation. The training age, speech intelligibility, fluency, communication, respiratory sound, cognate distinctions between voiced and voiceless sounds and intraesophageal pressures during phonation were significantly different between good and poor esophageal speakers. Vocal characteristics of good speakers differed significantly from those in the normal (P < 0.05 or P < 0.01). The upper intraesphageal pressure during esophageal phonation was higher than that of the normal, especially for poor esophageal phonation. The middle to lower intraesphageal pressures was highest during poor esophageal phonation. CONCLUSIONS: Esophageal phonation was completely alaryngeal, and its activators also differ completely, so phonation could not maintain much longer. Patients with poor esophageal phonation could not drive the air through the neoglottis freely. PMID- 12768787 TI - [Applied anatomy for the reinnervation of posterior cricoarytenoid muscle by phrenic nerve for bilateral vocal cord paralysis]. AB - OBJECTIVE: To study the anatomic basis for the anastomosis of phrenic nerve (PN) to the anterior branch of recurrent laryngeal nerve(RLN) for the treatment of the injured bilateral RLN. METHODS: The origin and the nutritive arteries and the adjacent tissue construction of PNs in 46 cases were studied. The longest utilizable length of PNs and the distance from the root of PN to cricothyroid joint were measured. The sectional area and the number of myelinated fibers of PNs and the anterior branch of RLNs were measured by computer image processing system. RESULTS: PNs coming from C4 comprised of 93.5%, 95.6% (44/46) of the nutritive arteries came from the ascending carotid artery and got into the cervical segment of PN from its root. The common trunk of PN was very deep, to the external of the common carotid artery and the vertebral vein, and deep to the internal jugular vein and thoracic duct (left), and in the superficies of the subclavian artery and in the deep of the subclavian vein when it was crossing the thoracic entrance. The distance from the root of PN to the level of the subclavian vein and to cricothyroid joint were (7.2 +/- 1.6) cm and (5.5 +/- 1.4) cm, respectively. The former was at least 1.5 cm longer than the latter. The average number of myelinated fibers and the sectional area of the PNs were 2.41 times and 2.15 times as many as those of the anterior branch of RLNs, respectively. The single-fasciculated PNs comprised of about 75.0% (18/24)). CONCLUSION: Clinically, it may be safe and available for cutting PN off at the level of the subclavian vein. The length of PN is enough for the anastomosis of PN to the anterior branch of RLN. PMID- 12768788 TI - [Lipopolysaccharide enhances DNA binding activity of nuclear factor-kappa B in the mouse cochlea]. AB - OBJECTIVE: To ascertain whether inoculation of lipopolysaccharide (LPS) into the mouse cochlea might induce enhancement of DNA binding activity of NK-kappa B and to establish a positive model of NF-kappa B activation in the cochlea. METHODS: Two groups of CBA mice were transtympanically injected either with 10 micrograms of LPS or an equal volume of saline. The nuclear proteins of the cochlea were extracted, quantitated and bound with 32P-radiolabeled oligonucleotide probe for NF-kappa B. The bound and free probes were separated by electrophoresis on a 4.5% native polyacrylamide gel. Supershift assays were carried out using the antibodies against NF-kappa B subunit P65 and P50. RESULTS: The radiolabeled probe bound differently to nuclear extracts from the LPS-treated and the saline treated mouse cochleae, showing that LPS inoculation significantly increased NF kappa B binding activity. Supershift analysis demonstrated that the enhancement of DNA binding activity was related to both P65 and P50 subunits. CONCLUSION: LPS could induce NF-kappa B activation in the cochlea. Moreover, this study implies that tissues from LPS-inoculated cochlea might serve as a useful positive control in the study on activation of NF-kappa B in cochlea. PMID- 12768789 TI - [A comparative experimental study between recombinant active gene 1-deficient mice and C57BL/6 mice model of acute bacterial rhinosinusitis]. AB - OBJECTIVE: To explore the infective course of acute bacterial rhinosinusitis in recombinent active gene 1 (Rag 1)-defecient mice (Rag1) and C57BL/6 mice (C57) and the difference between them after intranasal streptococcus pneumoniae inoculation. METHODS: Ten mice of each strain (Rag1 and C57) received Streptococcus pneumoniae strain T59, ATCC 49,619 suspended in trypticase soy broth, and controls (two mice for each strain) received trypticase soy broth alone. After 2, 5, 10 and 14 days, nasal lavage cultures were obtained and then the mice were killed. The heads were embedded with paraffin and serial sections were made for histological analysis. The percentage of sinus cavity occupied by neutrophil cluster (% cluster) and the number of polymorphonuclear leukocytes per square millimeter of sinus mucosa (PMN/mm2) were calculated by the use of a computer-aided microscope in conjunction with a reconstruction and image analysis system. RESULTS: % Cluster and PMN/mm2 in infected mice both of Rag1 and C57 appeared to peak on five and ten days separately, which were significantly heavier than those in controls(P < 0.05). The infection in C57 decreased by two weeks. But in contrast to C57, the infection in Rag1 had not been controlled and Streptococcus pneumoniae were still seen in the nasal lavage culture by two weeks. This difference between infected Rag1 and infected C57 was significant at P < 0.05. CONCLUSION: Acute bacterial rhinosinusitis in Rag1 and C57 mice were successfully induced by intranasal inoculation of streptococcus pneunoniae. This bacterial infection in C57 could be controlled completely and rapidly. In contrast, Rag1 failed to control rhinosinusitis and had a tendency to chronic inflammation, suggesting that T- and B-cell-dependent immunity was important for clearance of bacteria from rhinosinus and gene knockout mice was a convenient tool for investigation of the pathogenesis of experimental rhinosinusitis. PMID- 12768790 TI - [Significens of blood platelet CD62p in treating sudden deafness]. AB - OBJECTIVE: To compare different treatment methods for sudden deafness of different blood platelet CD62p levels. METHODS: Blood platelet was measured with flow cytometer. Patients with platelet CD62p over or less than 2% were treated with routine treatment (low molecular weight dextran, ATP, CoA and dexamethasone) or routine treatment plus batroxobin (thrombolysis) separately. RESULTS: In the group with platelet CD62p > or = 2%, the effective rates of routine treatment was 60.5%, and that of routine treatment plus thrombolysis was 84.6%. There was significant difference between them. In the group with platelet CD62p < 2%, the effective rates of two methods were 65.0% and 71.8% respectively(no significant difference between them). CONCLUSIONS: Detecting CD62p in sudden deafness patients makes sense for deciding treatment project. PMID- 12768792 TI - [Hypoxia effects on vascular endothelial growth factor derived epithelial cells of nasal polyps]. AB - OBJECTIVE: To understand the role of nasal mucous epithelial cells to hypoxia in early stage of nasal polyps(NP) formation. METHODS: Epithelial cells of NP and inferior turbinate (IT) were cultured without serum under normal oxygen and hypoxia, and stimulus of inflammatory cytokines. Erythropoietin (EPO) was regarded as hypoxia mark, and expression of vascular endothelial growth factor(VEGF) mRNA and protein derived from epithelial cells were detected respectively by in situ hybridization and ELISA. RESULTS: 1. Under hypoxia, EPO mRNA was expressed intensely in epithelial cells from NP and IT, and there was no significant difference between both of them. This result suggested that EPO might be regarded as a hypoxic mark. 2. The ability of producing VEGF mRNA increased with cytokines stimulation, especially under hypoxia. Protein level of VEGF from epithelial cells of NP and IT increased with cytokines stimulation, especially in hypoxia and was time-dependent. CONCLUSION: Epithelial cells actively produce vast VEGF under hypoxia. The VEGF induced by hypoxia of the mucosa in middle meatus is of importance in the formation of nasal polyps(NP) in early stage, which may be the major cause of NP formation in middle meatus. PMID- 12768791 TI - [The expression of matrix metalloproteinase-1 and interleukin-1-alpha messenger ribonucleic acid in human middle ear cholesteatoma]. AB - OBJECTIVE: To explore the expression of matrix metalloproteinase-1 (MMP1), tissue inhibitor of metalloproteinase-1(TIMP1) and interleukin-1-alpha messenger ribonucleic acid(IL-1 alpha mRNA) in acquired middle ear cholesteatoma, and to evaluate their roles in the molecular mechanisms of bone resorption. METHODS: Tissue specimens from 32 cases of acquired middle ear cholesteatoma and 14 cases of external ear skin were examined by immunohistochemical S-P method for MMP1 and TIMP1, and by in situ hybridization for IL-1 alpha mRNA. RESULTS: All 32 samples of cholesteatoma showed a stronger expression of MMP1 than the external ear skin. The integral absorbency of MMP1 in the two types of tissues were 2,018.26 +/- 174.89 and 1,428.35 +/- 123.39, respectively, with statistically significant difference between them. Neither of the two types of epithelial cells showed a remarkable expression of TIMP1. The cells hybridized for the antisence probes IL 1 alpha mRNA were only confined to the basal layer; in cholesteatoma, besides the basal cell layers, keratinocytes of the suprabasal cell layers were also found to contain specific hybridizations. The level of IL-1 alpha mRNA measured in cholesteatoma was significantly higher than that in the external ear skin. CONCLUSION: The overexpression of MMP1 and a derailment of the normally controlled MMPs-TIMPs system could play an active role in the molecular mechanisms of cholesteatoma invasion into the bone. The upregulation of IL-1 alpha might contribute to the increasing secretions of MMP1 in cholesteatoma. PMID- 12768794 TI - [Combined therapy of advanced tonsillar squamous cell carcinoma and one-stage repair of the defect]. AB - OBJECTIVE: To evaluate the treatment of advanced squamous cell carcinoma of the tonsil. METHODS: Eight patients with advanced tonsillar carcinoma underwent resections combining mandibular ramus and oral cavity approaches, neck dissection and one-stage repair of the defect with pectoralis major myocutaneous flap (PM). The managements were supplemented with preoperative chemotherapy and postoperative radiotherapy. RESULTS: Following up 3 to 9 years, 6 of 8 cases(75%) survived for more than 3 years. Two of 8 cases (25%) died of different causes postoperatively. One case died of serious hemorrhage resulted from radio mandibulomyelitis 3 months after operation, and the other case died of esophageal carcinoma 2 years after operation. Two cases complicated radio-mandibulomyelitis. Two cases suffered from temporary regurgitation of food to the nasal chamber. CONCLUSIONS: Through this combined approach, the advanced tonsillar carcinoma could be resected en bloc under direct visual field and keeping 1-1.5 cm safe margin to the tumor. Neck dissection, preoperative chemotherapy and postoperative radiotherapy were supplemental measures. Combined therapy was of great significance in reducing recurrence of the tumor. PM was usually survived and easily obtained, hence, suitable for repairing the pharyngeal defect. PMID- 12768795 TI - [Otorhinolaryngological neurofibromatosis]. AB - OBJECTIVE: To investigate the growth characteristic, clinical manifestation, diagnosis and treatment of neurofibromatosis (NF) in otorhinolaryngology. METHODS: Eleven cases of NF from 1989 to 2000 were analyzed retrospectively, among which, there were 5 and 2 cases occurring in nasal cavity and parapharynx (NF I), respectively, and 4 bilateral acoustic neuromas (NF II). RESULTS: NF I and NF II had long courses of 2-30 years, with common clinical features and unique characteristics. All patients with NF were operated on under general anesthesia. The NFs in the nasal cavity and sinus were resected through the paranasal approach and parapharyngeal NF through the lateral cervical approach. Unilateral acoustic neuromas were removed via the sigmoid sinus posterior suboccipital approach, one case bilateral acoustic neuromas was operated on in 2 stages. No death happened in this series. CONCLUSION: Surgical treatment is the best way for the treatment of NF. Radiotherapy can be considered in case of NF I with an extensive invasion, obscure border and recurrence. PMID- 12768793 TI - [Measurement of ostium area of maxillary sinus on three-dimensional imaging]. AB - OBJECTIVE: To measure the ostium area of normal maxillary sinus and maxillary sinus with inflammation on virtual endoscopy. METHODS: After scanning on coronal or axial position, data were transferred to workstation. Such anatomic structures as ostiomeatal complex (OMC) were viewed by virtual endoscopy (VE). Under the same condition, pixel/mm was worked out. RESULTS: Turbinates, meati and ostiomeatal complex were clearly viewed by virtual endoscopy(VE). The area of normal maxillary ostium was (16.45 +/- 1.21) mm2; the area of abnormal ostium was (18.28 +/- 2.12) mm2. The outside area of ethmoid infundibulum was (15.07 +/- 0.78) mm2 with normal maxillary sinus, and (17.19 +/- 1.59) mm2 maxillary sinus with inflamed. CONCLUSIONS: The areas of mormal and inflamed maxillary sinuses are equal, the outside areas of ethmoid infundibulum are unequal for normal maxillary sinus and maxillary sinus with inflammation. PMID- 12768796 TI - [Use of NiTi shape memory alloys in the eustachian tube to prevent and treat adhesive middle ear]. AB - OBJECTIVE: To explore and assess the use of NiTi shape memory alloys in the eustachian tube to prevent and treat adhesive middle ear. METHODS: All preliminary study cases were divided into three groups (A: 56 cases of adhesive otitis media, B: 27 cases of otitis media with effusion, C: 52 cases without ear disease as control). The eustachian orifice in all cases was observed by endoscopy. Autopsy of Eustachian tube was done in two specimens. The NiTi shape memory alloys were then installed into the Eustachian tube to prevent and treat adhesive middle ear in 4 cases. The installer of NiTi shape memory alloys was designed by ourselves. RESULTS: The Eustachian orifice can be classified into three types: wide, narrow and fissured. They comprised of 20, 31 and 5 cases in group A, 9, 13 and 5 cases in group B and 45, 7 and 0 cases in group C, respectively. The differences among three groups were statistically significant (A, B vs C, chi 2 = 29.66, 25.44 respectively, P < 0.001), but the difference between A and B was not statistically significant (chi 2 = 1.59, P > 0.05). Two cases with adhesive otitis media gained good ventilation with Valsalva method and the audiometric improvement of 15-30 dB after the NiTi shape memory alloys installation. One of the two cases with otitis media with effusion gained A-B gap closure over 8 months, and the other still improved with ventilative tube removed. In all 4 cases, there was no any side-effect or signs of patulous Eustachian tube. CONCLUSION: Adhesive otitis media can be prevented and treated by using NiTi shape memory alloys without any side-effect, but more study is necessary. PMID- 12768797 TI - [Analysis of randomized controlled trials on otorhinolaryngological diseases in China]. AB - OBJECTIVE: To evaluate the quality of randomized controlled trials (RCT) in otorhinolaryngology in China and offer evidence for the improvement of RCT. METHODS: Five kinds of Chinese journals of clinical otorhinolaryngology were searched, and RCTs were identified and analyzed according to the standards of Evidence Based Medicine. RESULTS: Two hundred and eighty seven issues were referred to, and eighty-one RCTs were finally identified and analyzed. Of these RCTs, 34.57% (28/81) had definite diagnostic standards, 38.27% (31/81) had including standards and 33.33% (27/81) had excluding standards; only 1.23% (1/81) got the approval of the participants; 40.74% (33/81) had moderate sample size, 3.70% (3/81) had large sample size and non of them mentioned sample size estimation; 81.48% (66/81) didn't report the method of randomization and 38.27% (31/81) had baseline comparison; 18.52% (15/81) didn't define the control interventions and 8.64% (7/81) even didn't explicate the experimental intervention; 32.10% (26/81) used blank comparison; 86.42% (70/81) didn't use blindness; 37.04% (30/81) didn't mention the adverse effects; 23.46% (19/81) used accredited standards to evaluate the outcomes; 11.11% (9/81) mentioned the loss of following-up and only 1.23% (1/81) treated the loss with statistics methods. CONCLUSION: The quantity and quality of otorhinolaryngological RCTs couldn't meet the clinical need. More high quality RCTs are required to improve the level of prevention and cure of otorhinolaryngologic diseases. PMID- 12768798 TI - [Vasodilators for sudden sensorineural hearing loss: a systematic review of randomized controlled trials]. AB - OBJECTIVE: To assess the effects and safety of vasodilators on sudden sensorineural hearing loss(SSHL). SEARCH STRATEGY: Electronic databases: MEDLINE from 1966, EMBASE from 1974, the Cochrane Controlled Trails Register, Chinese Bio medicine Database from 1989. Hand search: Five kinds of Chinese otolaryngology journals were selected. Literature references were checked intensively. SELECTION CRITERIA: Randomized controlled trials comparing vasodilators with placebo or other drugs in patients with SSHL. DATA COLLECTION AND ANALYSIS: Three reviewers independently accessed the quality of trials and extracted the data. RESULTS: Thirteen trials with 1,155 patients were eligible and covered in the systematic review. Ten of the trials were conducted in developed countries, and three in China. None of the four trials showed the effects of vasodilators on SSHL were better than that of placebo. None of the seven trials comparing different drugs showed the effects of one kind of vasodilators were better than that of the other vasodilators. Two trials showed that some other drugs, such as batroxobin and hypaque were probably better than the vasodilators (dextran, papaverine, 654-2, danshen). Eight trials showed side effects of vasodilators, such as pruritus, allergy, etc. CONCLUSIONS: Based on the systematic review of current eligible randomized controlled clinical trials, there is no evidence to prove that vasodilator therapy is more effective than placebo or other therapies for SSHL, or the effects of one kind of vasodilators to be better than that of the other vasodilators. So far we can't draw a reliable conclusion about the effects of vasodilators for SSHL. In addition, we must pay attention to their potential adverse reactions. PMID- 12768799 TI - [Ultrasonic prenatal diagnosis of congenital anomalies: the impact on medical management during pregnancy]. AB - Three women, aged 30, 18 and 37 years respectively, were diagnosed during the course of their pregnancies with congenital anomalies that carry a high risk of mortality and morbidity, namely a neural tube defect, gastroschisis and trisomy 22. All 3 women chose to continue their pregnancy and received various forms of counselling concerning the differences between the estimated prognoses. For the first patient, the emphasis was explaining the consequences of the disease for the unborn child and the parents and how the resulting handicaps could be minimised by medical treatment. For the second patient, the treatment plan was deliberately curative, and regrettably this failed. For the third patient, the attending physician guided the parents through their future loss and supported them during their period of mourning. PMID- 12768800 TI - [Evaluation of the Dutch Individual Health Care Professions Act]. AB - In 1997 the Dutch Individual Health Care Professions Act on the regulation of professions in health care came into force. This Act introduced a major reorganisation of the legal regulation of health-care professions. The evaluation of the Act, completed in 2002, indicates that the new legal structure has improved the quality of care. Yet several shortcomings have also been found in the Act, in terms of both the concept and the content. These are mainly due to discrepancies between the starting points and the concept of the Act on the one hand and the developments that have actually taken place in health care on the other hand, as well as a rather inactive system of inspection and control. The researchers who carried out the evaluation have formulated several proposals for improving the contents and effectiveness of the Act. PMID- 12768801 TI - [Youth health care: much prevention for little money]. AB - As part of government policy, the 'Youth healthcare' prevention programme is offered free of charge to all children aged 0 to 19 years who are resident in the Netherlands. It consists of a programme of primary prevention (including vaccinations, information and advice) and secondary prevention (screening, surveillance, early diagnosis) and individual prevention and care. Many elements from the programme package have been shown to have a favourable cost effectiveness relationship, in terms of health benefits and financially. Other elements have a social priority. The present government expenditure for the total youth healthcare package is about 380 million euros per year, that is 1900 euros per child. In terms of conditions prevented or years of life gained, this is cheaper than accepted prevention programmes for adults. The present approach can only be maintained and strengthened, if the expenditure is increased so that new programme elements can be investigated and--if found effective--implemented. PMID- 12768802 TI - [A welcome guide for evaluating medical research involving human subjects]. AB - In 1999, the Dutch Medical Research Involving Human Subjects Act came into force. The principal aim of this Act is to provide protection for human subjects who take part in medical research. Medical research involving human subjects may only be carried out after it has been approved by a recognised medical ethics committee. A central committee (Dutch acronym: CCMO) regulates the recognition of the local medical ethics committees and monitors their performance. For some types of research the protocol has to be reviewed by the CCMO itself. These are: non-therapeutic research involving children and mentally incompetent patients and research on gene therapy, xenotransplantation and embryos. The CCMO has recently published a guide for the local medical ethics committees. It presents an excellent overview of all the legal and practical aspects of the work of medical ethics committees. The guide is highly recommended. PMID- 12768803 TI - [Fetal surgery: for selected patients. The experiences of the Fetal Treatment Center in San Francisco]. AB - Foetal surgery, i.e. invasive treatment of the unborn child, has pros and cons. Foetal surgery is the therapy of choice in a selected group of patients. For obstructive uropathy, intrauterine urinary drainage is seldom indicated due to disappointing results. For congenital diaphragmatic hernia, the pressure in the respiratory tract is increased by tracheal occlusion therapy and so pulmonary hypoplasia is prevented. This probably has a beneficial effect for children with a poor prognosis. For congenital cystic adenomatoid malformation of the lung and foetal hydrops, resection of the lesion gives a survival rate of 70%. For sacrococcygeal teratoma and foetal hydrops, intrauterine resection of the tumour might save the life of the foetus. For high myelomeningocele, intrauterine covering of the defect has until now given no improvement in the neurological outcome. For twin transfusion syndrome, laser coagulation of the placental anastomoses is probably superior to serial amnioreduction. For women the mortality and morbidity associated with foetal surgery are low. PMID- 12768804 TI - [Surgical treatment of tumor metastases in the lungs, brain or liver]. AB - Metastases are generally an expression of widespread disease and therefore warrant systemic treatment. However, clinical observations have revealed that local surgical treatment might be beneficial in the case of organ-confined metastatic disease. Randomised studies have revealed that in the case of brain metastases, metastasectomy followed by radiotherapy, has a favourable outcome with respect to both the quality of life and overall survival. Retrospective non randomised studies in selected patient groups show prolonged post-treatment survival in the case of both lung and liver metastasectomy. The most important prognostic factors for metastasectomy are: disease control elsewhere in the body, tumour species, the patient's general condition, and the possibility of a total resection of the metastasis. These factors form the basis of the separate decision-making process for each individual patient. PMID- 12768805 TI - [New anticoagulants]. AB - A large number of newly developed platelet aggregation inhibitors and anticoagulants are currently being investigated in clinical studies. Most of these new agents are targeted to haemostatic pathways that have recently been shown to be of importance in vivo and usually have a higher efficacy than the currently available anticoagulants. The new platelet aggregation inhibitors can be divided into thienopyridine derivatives (ticlopidine, clopidogrel) and glycoprotein IIb/IIIa receptor antagonists (abciximab, eptifibatide, tirofiban). The new inhibitors of fibrin synthesis can be divided into direct thrombin inhibitors (hirudine, melagatran, ximelagatran), specific factor Xa inhibitors (pentasaccharides: fondaparinux, idraparinux) and inhibitors of the tissue thromboplastin factor VIIa complex (recombinant nematode anticoagulant protein c2, inactivated factor VIIa, recombinant tissue factor pathway inhibitor). In some cases this also results in a (relatively modest) increase in the risk of bleeding. The clinical use of the new compounds is often much more convenient than that of the presently available anticoagulants. PMID- 12768806 TI - [Diagnostic image (138). A girl with little bumps. Hand-foot-mouth disease]. AB - A 3-year-and-7-month-old girl with Down's syndrome had papules and vesicles on hands, feet and buttocks, and vesicular lesions in the oral cavity. A positive culture of enterovirus in vesicular fluid confirmed the clinical diagnosis 'hand foot-mouth disease'. PMID- 12768807 TI - [Mortality due to smoking in the Netherlands: 1.2 million tobacco-related deaths between 1950 and 2015]. AB - OBJECTIVE: To quantify the medical consequences of the tobacco use in the Netherlands for the past 50 years and the near future. DESIGN: Theoretical study based on the national death records and published risks by cause of death of tobacco use. METHOD: Observed lung cancer mortality (1950-1999) was related to birth cohort and age by a statistical model (according to Peto), and then projected into the near future. The smoking intensity was defined as the difference between the expected lung-cancer mortality if no one smoked and the observed lung-cancer mortality. Using this smoking intensity and published risks for other smoking-related causes of death, the model provided estimates of smoking-related mortality by age, sex and cause of death. RESULTS: In 1999, 18% (women) and 32% (men) of all mortality before the age of 70 was attributable to smoking. If no one had smoked, Dutch life expectancy for men and women would have been 3 years and 1 year higher, respectively. Between 1950 and 1999, 13% of all deaths were caused by smoking, the large majority (> 90%) of which occurred among men. Between 2000 and 2015, slightly more deaths are to be attributed to smoking (14%), 62% of which will be among men. In 2015, women will have caught up with men in terms of absolute numbers of lung-cancer mortality. CONCLUSION: Around one quarter of premature deaths were caused by smoking. In the near future, women of the baby-boom generation will have reached middle age and the highest (relative) smoking-related risks. It is important that clinical practice takes this increased risk of disease into account among middle-aged women who smoke. PMID- 12768809 TI - [Submental intubation: surgical and anesthesiological aspects]. PMID- 12768808 TI - [The efficacy of smoking cessation methods available in the Netherlands: a systematic review based on Cochrane data]. AB - OBJECTIVE: To obtain an overview of data from the Cochrane Library on smoking cessation methods and aids available in the Netherlands. DESIGN: Systematic literature review. METHOD: Common smoking-cessation methods in the Netherlands in 1999 and 2000 were selected from previous research. Data from relevant Cochrane reviews about these cessation methods were collected, after which the efficacy was calculated as a pooled odds ratio and the effectiveness as a percentage of 12 months' continuous abstinence. RESULTS: The following methods were found to be more efficacious than placebo: tailored written advice, individual counselling, telephonic counselling, group courses, all forms of nicotine-replacement therapy, bupropion and nortriptyline. Acupuncture was not superior to placebo. It was not possible to draw any unequivocal conclusions about hypnotherapy. No randomised studies were found with respect to the 'Allen Carr method'. Rates of 12 months' continuous abstinence were as follows for those methods with proven efficacy: tailored advice: 7%, individual counselling: 16%, telephonic counselling: 7.5%, nicotine gum: 17%, nicotine patch: 13%, nicotine inhaler: 17%, nicotine tablets: 20%, bupropion: 17%, and nortriptyline: 24%. The success rates for nicotine tablets and nortriptyline were based on only 2 and 1 study respectively. CONCLUSION: Several effective smoking-cessation methods are available in the Netherlands. In trials the long-term effectiveness of these methods was between 7 24%. PMID- 12768810 TI - [Overweight, obesity and underweigh in Switzerland: results of the 2000 Nutri Trend Study]. AB - The Nutri-Trend-Study 2000 conducted by Nestle Suisse SA and supported by the Swiss Federal Office of Public Health collected data on body weight and height of a random sample (n = 1004) of persons aged 18-54 years living in Switzerland. 3% of participants are underweight (BMI < 18.5 kg/m2), 26% overweight (BMI 25-29.9 kg/m2), 5% obese (BMI > or = 30 kg/m2) and 66% are of normal weight (BMI 18.5 24.9 kg/m2). Nutritional knowledge and attitudes of the BMI-groups do not differ substantially. All groups but especially participants with a BMI of 25+ should be better informed about the health consequences of an unbalanced diet. Furthermore they should be encouraged to adopt a physically active lifestyle and a healthy diet to remain within the BMI-range of normal weight or to reduce overweight. PMID- 12768811 TI - [Energy metabolism in long-term endurance sports: a case study]. AB - During the Swiss Citypower Gigathlon 2002, we determined energy turnover in one athlete. The average intake of calories was at 6500 calories per day and a deficit of 3500 calories per day resulted. This was due to the energy expenditure of about 10,000 calories per day. We concluded that it is extremely difficult or rather impossible to cover the total energy expenditure by nutrition during extreme events. PMID- 12768812 TI - [Blunt abdominal trauma in children. Traumatic intraperitoneal bladder rupture]. PMID- 12768813 TI - [Increased dyspnea. Mediastinal B-cell non-Hodgkin lymphoma with bilateral chylous pleural effusion]. PMID- 12768814 TI - [Anticoagulatin in atrial fibrillation--status of therapy in a network of physicians]. AB - Despite of sound evidence, that oral anticoagulation reduces substantially the incidence of stroke-events in patients with atrial fibrillation and concomitant risk factors, in daily practice only a part of these patients is in fact treated with anticoagulation. The aim of this cross-sectional study was to evaluate to which extent existing guidelines for the treatment of patients with atrial fibrillation are transformed in the "real world" context of a Swiss urban network of primary care providers. The greatest part (88.4%) of the included patients with atrial fibrillation had a high risk for thromboembolism and herewith the indication for anticoagulation was given. For a primary care collective the amount of patients with anticoagulation was high: 74.1% did receive this kind of preventive therapy. 89% had robust International normalized ratio (INR) values within the recommended range (INR 2.5-3.5). The most common reasons not to prescribe oral anticoagulation were: old age (17.5%; mostly in combination with other reasons), risk of falls (16.5%), medical indication not given (16.5%). Thus the recommendations of the guidelines have been transformed into practice for a bigger part of the included patients. Given these results a broadly implemented quality-improvement-project to bring up the anticoagulation rate even higher hardly seems to be efficacious and is presently not of top priority in this network of primary care providers. PMID- 12768815 TI - Antibiotic-associated diarrhea: therapeutic aspects and practical guidelines--an interdisciplinary approach to a common problem. AB - Antibiotic-associated diarrhea (AAD) is a common complication of antibiotic treatment, most often seen in non-hospitalised patients. In principle, such diarrhea can be triggered by any antibiotic. An interdisciplinary working group discussed the different aspects of AAD in view of its gastroenterological, microbiological, paediatric, general medical and pharmaceutical implications, also in consideration of the position of patients and health insurance funds. This paper implies therapeutic aspects and practical guidelines to raise awareness of these problems also in routine situations and to enable the persons and institutions involved on the various levels of the health-care system (patients, pharmacists, family doctors, specialists and hospitals) to handle the problem of AAD more easily in a standardised way as far as diagnostics, therapy and prevention are concerned. PMID- 12768816 TI - [Vertigo and arterial hypertension in younger patients. Hyperaldosteronism (Conn syndrome) in diastolic arterial hypertension, chronic hypokalemia, right adrenal gland adenoma]. PMID- 12768817 TI - [Non-specific influenza symptoms and microhematuria. IgA nephritis/Berger disease]. PMID- 12768818 TI - [The task remains]. PMID- 12768819 TI - [International Day of Nurses]. PMID- 12768820 TI - [The long struggle against the virus]. PMID- 12768821 TI - [Residential facilities for better quality of life]. PMID- 12768822 TI - [New structures as chance: the new clinic aat Ludwigsburg-Bietigheim. New problems at the ward management level]. PMID- 12768823 TI - [Priorities. Sitting is not always simple]. PMID- 12768824 TI - [Successful outcome by the German Nursing Society--follow up of quality management. The result of clinical management]. PMID- 12768825 TI - [Knowledge and action]. PMID- 12768826 TI - [Is "sister" still the appropriate term?]. PMID- 12768827 TI - [Kaposi's sarcoma]. PMID- 12768828 TI - [Characteristics of Kaposi's sarcoma. A retrospective study in a reference hospital]. AB - Kaposi's angiosarcoma (SK) is a vascular tumour that affects skin and other organs. Nowadays there is thought that immunosuppression is one of the factors related with its genesis. OBJECTIVES: Show the information corresponding to the distribution of the SK, proportion of subtypes, evolution and therapeutic used. MATERIAL AND METHODS: A 28 cases retrospective study. We obtained the personal background, habits, origin of the VIH when the SK was associated with AIDS, and treatment carry out. Type of SK and location, cutaneous presentation, complications and B symptoms. Treatment, response and adverse effects. Progression of the disease, average time of relapse and survival, and presence of the SK at death. RESULTS: 64% epidemic, 21% classic type and 14% associated with transplants. In 48% of the epidemic cases, the VIH assumed to homosexual habits and 18% to being ADVP. In all there was cutaneous affectation, 46% had affectation of mucouses, 11% adenopathies and 14% visceral extension. 71% received treatment: 40% chemotherapy, 35% radiotherapy and 18% cryotherapy. Progression: 11% became stable, in 50% there was local extension and systemic in 17%. The average survival was of 4.16 +/- 3 years. CONCLUSIONS: SK has a minor incidence in the homosexual population. The high proportion of classic and transplants associated affirm the role of immunosuppression as a predisposing factor. There were numerous local complications that were associated with B symptoms. The cryotherapy was used as the first option in the located forms and in the widespread ones was the chemotherapy. PMID- 12768829 TI - [GB virus C: lack of association with transaminases levels, CD4 and HIV viral load in aids patients]. AB - OBJECTIVE: To study the prevalence of GBV-C-RNA in sera of HIV-infected patients and determine whether differences in immunological condition and hepatic disease exist between GBV-C positive and negative patients. METHODS: The presence of GBV C-RNA was determined in sera of 222 HIV-positive patients by semi-automated RT PCR. A comparison of GBV-C-RNA positive and negative patients was made by studying a series of clinical and analytical parameters. This same comparison was made in particular between those coinfected with HCV and GBV-C and those who only presented GBV-C. RESULTS: Prevalence of GBV-C-RNA was 28.8%. The most frequent hepatotropic virus was HCV, appearing in 71.6% of cases. Coinfection with HCV and HGV was present in 17% and 8.6% only had GBV-C. Patients positive for GBV-C-RNA showed clinical and analytical characteristics similar to those found in GBV-C RNA negative patients. Among the HCV-GBV-C coinfected and those presenting HGV as the only virus it was observed that the coinfected group presented alterations in transaminases and predominance of parenteral transmission as a risk factor for HIV, whereas the GBV-C group presented normal transaminases and predominance of sexual transmission. No differences were perceived in mean CD4 and HIV-RNA values in both groups. CONCLUSIONS: Being positive for GBV-C in HIV-positive patients does not influence the presence of hepatic disease that in these patients is frequently accompanied by coinfection with other hepatotropic viruses. Moreover, it does not seem to influence the viremia of the HIV nor the CD4 cell counts. PMID- 12768830 TI - [Utility of anaerobic blood cultures in extra-hospitalary bacteremias]. AB - OBJECTIVE: To compare the rentability of anaerobic hemocultures with aerobic cultures in patients suffering bacteremia, trying to analyze the rentability of anaerobic cultures. METHODS: There were analyzed all the hemocultures taken by medical decision for one month. There were taken three extractions by patient in different venopunctures points. In each extraction was taken 10 ml (5 ml in aerobic bottle, and 5 ml in anaerobic bottle). There were compared microorganism isolated in aerobic bottles and anaerobic bottles. Three physician analyzed all the clinical dates of the patients with positive hemocultures and classified them as negatives, contaminants and significant. RESULTS: There were taken hemocultures of 180 patients. There were analyzed 152 hemocultures groups (3 extractions aerobic-anaerobic), without studying 28 owe to not following the study protocol 24 (13.3%) or because of not possibility of studying clinical story 4 (2.2%). Aerobic extractions: Negatives 91 (59.86%), contaminants 36 (23.68%), significant with or without contaminant 25 (16.44%). Anaerobic extractions: Negatives 127 (83.5%), contaminants 2 (1.31%); significant with or without contaminant 23 (15.13%). There are 6 (8.6%) significant positive isolated in anaerobic bottles, that were negative in aerobic bottles, and are: 3 E. coli, 1 K. pneumoniae, 1 Peptostreptococcus sp, 1 Bacteroides sp, and over the total significant isolated (31) are the 19.35%. Both anaerobic bacteremias were clinically suspected. Contaminant microorganism more frequently isolated in aerobic bottles was Staphylococcus sp, 25 cases (69.44% of all contaminants). Significant microorganism more frequently isolated was E. coli (in anaerobic bottles too) in 12 cases (38.70%) of all significant isolated, following by Streptococcus pneumoniae with 4 cases (12.9%). CONCLUSIONS: We recommend collection of one aerobic and one anaerobic blood culture bottle per blood culture set because the global rentability of hemocultures is increased significantly. PMID- 12768832 TI - [Osteonecrosis and HIV]. AB - An increase of the incidence of osteonecrosis in subjects infected with the HIV has been reported. It is unknown if osteonecrosis is an HIV infection consequence, or just the consequence of so many risk factors present among HIV infected subjects. We report five osteonecrosis cases found in our series of 534 HIV patients, and we review the related bibliography. All cases have some conventional risk factor different to the HIV or the antiviral treatment. We can't conclude antiviral treatments are not involved in osteonecrosis, but they have been the only risk factor in none of our patients. PMID- 12768831 TI - [Respiratory symptoms and lung function in a geriatric population of a Galician rural community: a pilot study]. AB - INTRODUCTION: Respiratory diseases are a frequent cause of health demands and have a large impact on morbidity and mortality of the Galician population, especially among the older one. Recent work shows that the diagnosis and treatment of these diseases is not optimal. This increases the utilisation of health care resources. MATERIAL AND METHODS: We studied 28 patients of the municipality of Val del Dubra (Northwest Spain) aged between 65 and 74 years. We performed a spirometric exploration and carried out a questionnaire interview on respiratory symptoms, life style, and occupational and health-related antecedents. RESULTS: Among men, 54% of were or are smokers. None of the women ever smoked. Respiratory symptoms were more frequent among women than among men (80% versus 54%). In the spirometric study, the largest volumes and flux are observed among non-smoking males who do not report dyspnea. DISCUSSION: Respiratory symptoms are frequent in the rural population aged between 65 and 74 years. Tobacco consumption is similar to other Spanish communities, but different from that seen in other countries. Male gender, non-smoking status and absence of respiratory symptoms are associated with higher spirometric figures. PMID- 12768833 TI - [Long-term treatment with low molecular weight heparin, of chronic disseminated intravascular coagulation]. AB - The term "disseminated intravascular coagulation" (DIC) defined a pathologic process which complicates the clinical course of many diseases; it is characterized by huge amounts of thrombin and plasmin within the circulation. There are a lot of causes of these intermediary mechanism of disease, among these, infections and neoplasia are the most frequent. Aortic aneurysm is a vascular disease than can be complicated with DIC. We report a case of a patient affected of chronic disseminated intravascular coagulation complicated with systemic hemorrhagic syndrome, of vascular origin (an aortic aneurysm). It was treated with a low molecular weight heparin (LMWH), but in the presence of an allergy disorder the drug was discontinued and substituted by another LMWH. The hemorrhagic complications were treated with antifibrinolytics associated to the LMWH. The drug was held up 30 months with an acceptable performance status and no significant secondary effects except osteoporosis. PMID- 12768834 TI - [Forearm malignant epithelioid schwannoma associated with melanoma]. AB - Tumours originating in peripheric nerves usually appear in patients with neurofibromatosis (NF) signs, presenting frequent combinations of tumours in nerves and cutaneous lesions. Nevertheless, this association is very rare in cases without NF. Therefore, the aim of the present article is to present a case of malignant melanoma and malignant schwannoma, without any described NF. A 69 year-old woman with antecedents of malignant melanoma diagnosed two years previously in the dorsum of the fifth finger of the left hand, treated by means of amputation of the finger. The patient presented a malignant epithelioid schwannoma adhered to the median nerve that required elbow amputation. One month later lung metastases appeared and the patient died. This case presented no known NF sign. However, a relationship must be searched for in the common origin of melanoma and schwannoma from the embryonic neural crest. PMID- 12768835 TI - [Iliopsoas abscess and systemic lupus erythematosus]. AB - Iliopsoas muscle abscess (IPA) is an uncommon condition, and it is usually associated with immunosuppression. Three out of a cohort of 552 patients diagnosed of systemic lupus erythematosus (SLE) developing an IPA, are reported herein. Patients showed fever and other symptoms related to SLE. They improved only partially under SLE therapy, and showed pain suggestive of IPA. It was confirmed by CT in all cases. S. aureus was isolated in one patient (primary IPA), and M. tuberculosis in the others. Specific antimicrobial therapy and surgical drainage were required. In summary, SLE might be considered as a risk condition for the development of IPA, due to the immunosuppression inherent in the disease and its treatment. PMID- 12768836 TI - [Regulation of bronchial tone in chronic obstructive pulmonary disease (COPD): role of muscarinic receptors]. AB - Anticholinergic agents have proved to be of particular value in the treatment of COPD, as vagal cholinergic tone appears to be the only reversible component of airway narrowing, opposite to what happens in asthma. Anticholinergics block muscarinic receptors on airway smooth muscle an submucosal gland cells. Three subtypes of muscarinic receptors have been demonstrated in human airways. M1 receptors in parasympathetic ganglia facilitate cholinergic neurotransmission. M3 receptors on airway smooth muscle cells and glands mediate bronchoconstriction and mucus secretion. M2 receptors at cholinergic nerve endings inhibit the release of acetylcholine and therefore act as feedback inhibitory receptors (autoreceptors). Ipratropium bromide is a non-selective muscarinic antagonist and therefore blocks M1, M2 and M3 receptors. Tiotropium is a novel, potent, and long lasting muscarinic antagonist that has a kinetic selectivity for M1 and M3 receptors because it dissociated very quickly from M2 receptors. Once-daily inhaled tiotropium is a safe and effective bronchodilator useful as a first-line maintenance therapy in COPD. PMID- 12768837 TI - [Psychiatric manifestations due to most common endocrine diseases]. PMID- 12768838 TI - [Genetics of iron overloads and hereditary hyperferritinemia cataract syndrome]. PMID- 12768839 TI - [Diffuse malignant pleural mesothelioma with unusual radiological presentation]. PMID- 12768840 TI - [Lower gastrointestinal bleeding by Dieulafoy colonic ulcer in patient with mitral valvulopathy and atrophic chronic gastritis]. PMID- 12768841 TI - [Pulmonary edema and renal failure in cocaine (crack) abuser patient]. PMID- 12768842 TI - [RS3PE syndrome as a first manifestation of prostatic carcinoma]. PMID- 12768843 TI - [Necrotic-haemorrhagic lesions in human extremities caused by meningococcus sepsis]. PMID- 12768845 TI - [Mesenteric thrombosis, hyperhomocysteinemia and oral contraceptive agents intake]. PMID- 12768844 TI - [Thoracic pain and ulcerous colitis]. PMID- 12768846 TI - [Lung adenocarcinoma, inflammatory bowel disease and azathioprine]. PMID- 12768847 TI - [Non-bloody pleural effusion: a rare complication of percutaneous liver biopsy]. PMID- 12768848 TI - Does the doctor still have the right to make mistakes in 2003? PMID- 12768849 TI - Patient's rights: a figment for patients, a threat to surgeons! PMID- 12768850 TI - The medical responsibility: current view from the Council of Physicians side. AB - The medical responsibility has been clearly defined in the Royal Decree no. 78 dated November 11, 1967 concerning the medical practice. Moreover, several articles from the Ethical Code (Code de Deontologie) have clarified some social and economical responsibilities in the medical practice (articles 99 to 103) and the quality of patient care (article 36). The National Council has also published at least 31 advises facing the daily reality and the growing insecurity. That atmosphere is coming from the jurisprudence, the increasing responsibility insurance fees, the obligation of results instead of means, and the project of patient rights law. That project is currently dissociated from other projects such as an update on the medical responsibility and/or the no fault indemnity. Therefore, there is a current need for developing written patient information and using informed consent forms for risky surgical procedures. Before recognizing the no fault concept with indemnity, it is necessary to review the coverage of the responsibility insurance, educate the medical doctors in the no fault concept, study the mode of compensation for therapeutic hazards and differentiate the objective and subjective parts of the patient's chart. PMID- 12768851 TI - [Medical responsibility: recent development of the jurisprudence]. PMID- 12768852 TI - [Medical responsibility: the new law relative to patients' rights]. PMID- 12768853 TI - [Medical Responsibility and no fault]. PMID- 12768854 TI - Surgical management of bile duct stricture following laparoscopic cholecystectomy. PMID- 12768855 TI - Prevalence of bile duct injury following cholecystectomy. PMID- 12768856 TI - Prevention of bile duct injury (BDI) during cholecystectomy. PMID- 12768857 TI - Bile duct injury during laparoscopic cholecystectomy: risk factors, mechanisms, type, severity and immediate detection. PMID- 12768858 TI - The role of intraoperative cholangiography in detecting and preventing bile duct injury during laparoscopic cholecystectomy. PMID- 12768859 TI - [Medical Responsibility and laparoscopic surgery]. PMID- 12768860 TI - National survey on cholecystectomy related bile duct injury--public health and financial aspects in Belgian hospitals--1997. AB - Public health and financial aspects of cholecystectomy related bile duct injury (BDI) are highlighted in a National Cholecystectomy Survey carried out through 'datamining' the Federal State Medical Records Summaries and Financial Summaries of all Belgian hospitals in 1997. All cancer diagnoses, children < or = 10 years, cholecystectomies performed as an abdominal co-procedure or patients having undergone other non-related surgery were excluded from the study. 10.595 laparoscopic (LC) and 1.033 open cholecystectomies (OC) as well as 137 secondary BDI treatments (LC/OC) were included in the survey (total 11.765). Both LC and OC groups turned out to be significantly different as to distribution of patient's age and APR-DRG severity classes. Composite criteria in terms of ICD-9-CM and billing codes were elaborated to classify: 1) primary, intra-operatively detected and treated BDI (N = 30), 2) primary delayed BDI treatments (N = 38), 3) secondary BDI treatments (N = 137), 4) non-BDI abdomino-surgical complications (N = 119), 4) uneventful laparoscopic (N = 7.476) and 5) uneventful open cholecystectomy (N = 681). Complication rates, community costs of LC and OC groups, incidence of preoperative ERCP and/or intra-operative cholangiography as well as interventions for complications were studied. Incidence of cholecystectomy related BDI was 0.37% in LC, 2.81% in OC and 0.58% overall. Average costs amounted to [symbol: see text] 1.721 for uneventful LC, [symbol: see text] 2.924 for uneventful OC, [symbol: see text] 7.250 for primary, intra operatively detected and immediately treated BDI [symbol: see text] 9.258 for primary delayed BDI treatments, [symbol: see text] 6.076 for secondary BDI treatments and [symbol: see text] 10.363 for non-BDI abdomino-surgical complications. In conclusion BDI with cholecystectomy reveals to be a serious complication increasing the overall average cost factor ninefold if not detected intra-operatively, in which case the raise is only fourfold. As a consequence BDI should be avoided by all means. In this respect 4 crucial surgical guidelines are emphasised. PMID- 12768861 TI - National Survey of Bile Duct Injury Associated with Cholecystectomy. Big Brother in the operating room versus another EBM tool for critical assessment of surgical innovation? PMID- 12768862 TI - Two decades of research on etiology and genetic factors in the abdominal aortic aneurysm (AAA)--with a glimpse into the 21st century. AB - Long gone are the days when the question of "etiology" of the abdominal aortic aneurysm (AAA) required a simplistic response. AAA is caused by an interplay of environmental and genetic factors, each of which may modify the expression of the other. A low penetrance, an insidious onset, and a wide distribution have forced scientists to rely on complex approaches to elucidate the pathophysiology of AAA. Hypotheses over the last twenty years have evaluated several components of connective tissue structure, function, and regulation. Although there has been considerable overlap in the many genetic approaches undertaken to explain aneurysm development, research recently has focused on reverse-engineering techniques. While earlier investigations using a "forward" or "candidate gene approach" have provided many insights in the understanding of AAA disease, advances in statistical modeling, new techniques in molecular biology, and gross computing power have now made more feasible the development of a reverse approach. New hope lies with the development of biochemical and computing tools which have paralleled information's vast dissemination in this rapidly widening field. Aortic inflammation and the upregulation of proteases and dysfunctional tissue-turnover are receiving more attention. No doubt, the interplay of all of these technologies will occupy investigators with an interest in AAA etiology for many years to come. PMID- 12768863 TI - What are the long-term results of conventional open surgical repair of abdominal aortic aneurysms? AB - PURPOSE: To determine the long-term results of conventional open surgical repair of abdominal aortic aneurysms (AAAs) and the prevalence of late arterial abnormalities. METHODS: CT scan follow-up was obtained between 8 and 9 years after elective AAA repair on a cohort of patients enrolled in the Canadian Aneurysm Study, a registry that originally consisted of 680 patients. A request for CT follow-up was sent to the responsible surgeon in 1994 when 251 patients were alive and available. Ninety-four of the 251 patients agreed to undergo an abdominal and thoracic CT scan, and each scan was interpreted independently by two vascular radiologists. RESULTS: The aorta was analysed in five defined segments, and an aneurysm was defined as > 50% enlargement from the expected normal value as defined in the Reporting Standards for Aneurysms. Using this strict definition, 64.9% of patients had an aneurysm, but the abnormality was considered a possible indication for surgical repair in 13.8%. Of the 39 patients who had an initial repair with a tube graft, 12 (30.8%) were found to have an iliac aneurysm and 6 (15.4%) were considered to be of possible surgical significance. The median graft size at the time of operation was 18 mm, which increased to a median size of 22 mm at follow-up. Fluid or thrombus around the graft was observed in 28%, and bowel was intimately associated with the graft in 7%. CONCLUSIONS: The longterm results of conventional open surgical repair is durable. CT scan follow-up between 8 and 9 years postoperatively often demonstrates aortic and iliac abnormalities, but the majority are not clinically significant. On the basis of these findings, a routine CT scan of the abdomen and chest is recommended after 5 years. This study provides a population based study for comparison with the longterm results of endovascular repair. PMID- 12768864 TI - Laparoscopic hand-assisted abdominal aortic surgery for aneurysmal and occlusive disease: a one-year clinical experience. AB - PURPOSE: To describe our technique and one year clinical experience with laparoscopic hand-assisted abdominal aortic surgery for aneurysmal and occlusive disease. PATIENTS AND METHODS: Between June 2001 and May 2002, 13 patients underwent a laparoscopic hand-assisted abdominal aortic operation. RESULTS: The median operating time and aortic cross-clamp time were 230 and 29 minutes respectively. The median intra-operative blood loss was 625cc. There were no peri operative (30 day) deaths. The conversion to open surgery ratio was 1/13. One major arterial and one systemic complication occurred. Median hospital stay was 6 days (range 4-42 days). CONCLUSION: Laparoscopic hand-assisted abdominal aortic surgery for aneurysmal and occlusive disease is technically feasible and safe. The use of this laparoscopic technique combines the advantage of minimally invasive surgery and the reliability of an established open procedure without the risk of endoleaks in aneurysm surgery. PMID- 12768865 TI - A comparison between minilaparotomy and standard median laparotomy for reconstruction of aorto-iliac occlusive disease. AB - OBJECTIVE: Minimally invasive surgical procedures have become increasingly used in all surgical branches. In this respect we compared the minilaparotomy (ML) technique with standard median laparotomy (SML) for the surgical treatment of aorto-iliac occlusive disease. METHODS: 120 patients were included in this prospective study, with 60 patients in the minilaparotomy group and 60 patients in standard median laparotomy group. The two approaches were used within the same time period. Both groups were compared in terms of operating and aortic cross clamp time, the amount of blood transfusion, length of stay in the intensive care unit and hospital, and time for normalization of bowel functions. RESULTS: There were no significant differences in terms of the amount of blood transfusion, or aortic cross clamp time. The minilaparotomy approach has the advantages of less operating time, earlier return to a general diet, decreased length of stay in the intensive care unit and hospital and reduced cost. CONCLUSIONS: As a conclusion, we believe that the minilaparotomy technique will be the standard treatment of choice for aorto-iliac occlusive disease in the future with reduced use of facilities and lower cost than the laparoscopic or the standard technique. PMID- 12768866 TI - Arm morbidity after complete axillary lymph node dissection for breast cancer. AB - The aim of the study was to clarify the factors causing and/or influence morbidity following axillary dissection in patients treated for breast cancer by either lumpectomy or mastectomy. The records of 106 women with invasive breast cancer treated between 1996 and 1997 were retrospectively reviewed. Objective assessment included measurement of lymphoedema, shoulder mobility and axillary sensation. A questionnaire was used for subjective assessment of arm morbidity and pain. Lymphoedema was present in 13% of patients, a restriction in shoulder function in 24%, while 93% of patients had an impaired sensation in the axillary region. Lymphoedema and restriction in shoulder function were common in patients after adjuvant axillary radiation. Morbidity following axillary lymph node dissection is high and confirms the potentially severe effects of a staging procedure on a relatively young population. Adjuvant radiotherapy increases morbidity significantly and therefore indications for adjuvant axillary radiotherapy should be revised with scrutiny for each patient individually, bearing in mind the disastrous consequences of the combination of radiotherapy and surgery on the axilla. PMID- 12768867 TI - Chylous leak: a rare complication after axillary lymph node dissection. AB - The authors present a case of a chylous leak after axillary lymph node dissection. Although this case has been treated conservatively, the authors discuss different treatment approaches. PMID- 12768868 TI - Peri-areolar skin-sparing mastectomy and immediate tram flap reconstruction. AB - The association of peri-areolar skin-sparing mastectomy with immediate autologous tissue breast reconstruction could be proposed to selected patients necessitating mastectomy to control their breast cancer. A prospective series of 14 consecutive patients were operated using this association and evaluated. Mean age at time of surgery was 52 years. Mean operating time for reconstruction was 163 minutes. There were two minor local complications, one needing secondary adaptation under local anaesthesia. Cosmetic result was judged good to excellent by all the patients with a mean follow-up of 13.2 months. PMID- 12768869 TI - Symptomatic adenomyomatosis of the gallbladder--report of a case. AB - Adenomyomatosis of the gallbladder is a benign and degenerative condition of the gallbladder, characterized by proliferation of the mucosa of the gallbladder wall, forming invaginations and diverticula, penetrating a thickened muscular layer: the so-called Rokitansky-Aschoff sinuses (RAS). Most of the patients with adenomyomatosis remain asymptomatic. Hence adenomyomatosis is usually an incidental finding, either on ultrasonography performed for the detection of stones or by histologic examination of surgical gallbladder specimens. Only occasionally does adenomyomatosis not associated with cholelithiasis cause right upper quadrant pain. We report a case of symptomatic adenomyomatosis of the gallbladder. Clinical findings, etiology, diagnosis and therapy are discussed. PMID- 12768870 TI - Emphysematous cholecystitis. AB - Emphysematous cholecystitis is a relatively rare variant of acute cholecystitis with infection by gas-producing organisms. Diagnosis involves the demonstration of gas within the lumen or wall of the gallbladder by ultrasound or CT scan. In contrast to acute cholecystitis, emphysematous cholecystitis occurs more commonly in elderly and diabetic patients, and is frequently associated with perforation and death. We report here a case of a 75-year old man who developed emphysematous cholecystitis. PMID- 12768871 TI - Metastatic carcinoma of the gallbladder after a renal cell carcinoma. AB - The gallbladder is rarely the site of distant metastases and in most cases malignant melanoma is the primary tumor. We report a case of a 64-year-old man with a gallbladder metastasis secondary to a renal cell carcinoma. Renal cell carcinoma has a tendency toward metastatic disease, the most notable features of this tumor being its unusual pattern of metastatic disease. Pre-operative imaging studies are often futile in the differentiation between primary and secondary tumors of the gallbladder. Since primary tumors of the gallbladder often coexist with gallstones, a polypoid lesion in an acalculous gallbladder is more consistent with metastasis than a primary tumour. If feasible, surgical resection of the gallbladder is mandatory because it could guarantee better chances of survival for patients with metastatic renal carcinoma. PMID- 12768872 TI - Congenital epulis of the newborn. PMID- 12768873 TI - How to perform a below-knee amputation. AB - Even though modern techniques have improved patient survival and limb salvage rates in patients with critical limb ischaemia and end-stage vascular disease, amputation is sometimes the only possible treatment. In younger patients with traumatic avulsion of a foot, infected gangrene of the foot or a peripheral tumour, amputation is out of discussion and commonly accepted. In older vascular patients, amputation should rather be considered as the starting point for revalidation and rehabilitation than as failure of a revascularization technique. The evolution in prostheses permits a rapid revalidation in most patients. However, an accurate amputation technique is still required to produce a good quality stump allowing early fitting of prosthetics. PMID- 12768874 TI - [Is it possible to have organized screening for ovarian cancer?]. PMID- 12768875 TI - Cytogenetic alterations in liver cell tumors as detected by comparative genomic hybridization. AB - Hepatocellular carcinoma (HCC) is one of the most frequent neoplasia worldwide. Environmental risk factors as hepatitis B virus (HBV) and hepatitis C virus (HCV) play a critical role in liver carcinogenesis. Liver carcinogenesis is probably a multistep process involving several genes, but morphological and molecular features of premalignant and malignant hepatic lesions are yet far from being fully elucidated. This study summarizes chromosomal imbalances of a wide variety of precancerous and cancerous lesions of the liver as detected by Comparative Genomic Hybridization (CGH). Preneoplastic nodules, classified as macroregenerative nodules (MRN), low-grade dysplastic nodules (LG-DN) and high grade dysplastic nodules (HG-DN), showed only few aberrations (mean 1.1/case), without any significant pattern. This finding is comparable to what happens in non-neoplastic tissue. On the contrary, in three of six HG-DN we found deletions of 8p and gains of 1q. LG-DN and MRN never showed these chromosomal imbalances. Furthermore some chromosomal changes appeared to be quite characteristic for some of the investigated tumor entities: HCC. 8p- might be relatively specific for HCC (44% with 8p) since it was hardly found in fibrolamellar carcinoma (FLC) or hepatoblastoma (HB). The same applies for 17q+ which was much more frequent in HCC (41%) than in FLC (0 of 5) or HB (9%). FLC. There were 4 alterations that were more frequent in FLC than in other tumor types. These changes included 4q+, 9p-, 16p- and Xq-. HB. Alterations that were typical for hepatoblastoma included Xp+ and Xq+, which were found both in the mesenchymal and in the epithelial parts of HBs. Other frequent changes in these tumor types included 1p-, 9p- and 2q+. PMID- 12768876 TI - [Atypical, aplastic, and unusual meningiomas. Morphology and incidence in 300 consecutive cases]. AB - We morphologically studied 300 consecutive and primitive meningiomas surgically treated between march 1997 and april 2002 in order to evaluate the incidence of atypical, anaplastic, and morphologically unusual meningiomas. Two hundred and fifty-five meningiomas (85%) were WHO I, 33 (11%) were WHO II, 9 (3%) were WHO III; the remaining 3 meningiomas (1%) showed clear and diffuse oncocytic differentiation without cytologic or architectural atypia (oncocytic meningiomas). Forty-five of 255 WHO I meningiomas (18%) were infrequent histological subtypes: 18 (7%) psammomatous, 9 (4%) metaplastic, 9 (4%) secretory, 6 (2%) angiomatous, and 3 (1%) microcystic. Thirty of 33 WHO II meningiomas (91%) were atypical, 2 (6%) were clear cell meningiomas, and 1 (3%) was chordoid meningioma. Seven of 9 WHO III meningiomas (78%) were anaplastic and 2 (22%) were papillary. We evidenced the high morphological variability and the discrete occurrence of WHO I and WHO II meningiomas. PMID- 12768877 TI - DNA ploidy in cervical lesions assessed by computed image analysis: relation to histopathology. AB - OBJECTIVE: To assess the different patterns of DNA nuclear content in cervical intraepithelial neoplasia. STUDY DESIGN: DNA nuclear content patterns (DNA ploidy) were assessed in 82 paraffin embedded samples diagnosed as chronic cervicitis CIN 1, CIN 2 and CIN 3 by image analysis using the CAS200 BD System. These results were compared to the consensus diagnosis of two pathologists. The chi-square test was used to evaluate the results of classificatory variables (CIN 1, CIN 2 and CIN 3). RESULTS: Aneuploidy was found in 86% of CIN 3; 59% CIN 2; 26%% CIN 1. All samples classified as chronic cervicitis were diploid. CONCLUSIONS: The DNA nuclear content was found to parallel the three grades of the histological WHO classification. Aneuploidy was strongly associated with the high-grade lesions; the detection of a subset aneuploidy of low grade lesions would suggest a indicate closer follow-up of these cases. PMID- 12768878 TI - [Limitations of colonic lavage in the cytopathological diagnosis of inflammatory and neoplastic lesions]. AB - The authors describe the personal experience on colonic lavage cytology in neoplastic or inflammatory diseases. Although the presence of false negative results in the lesions of right colon, the method could be useful in the diagnosis of selected cases. PMID- 12768879 TI - [Fractal analysis of liver fibrosis]. AB - This study realized by two different study groups use of Fractal geometry to quantify the complex collagen deposition during chronic liver disease. Thirty standard needle liver biopsy specimens were obtained from patients with chronic HCV-related disease. Three mu-thick sections were cut and stained by means of Picrosirius stain, in order to visualise collagen matrix. The degree of fibrosis was measured using a quantitative scoring system based on the computer-assisted evaluation of the fractal dimension of the deposited collagen surface. The obtained results by both study groups, show that the proposed method is reproducible, rapid and inexpensive. The complex distribution of its collagenous components can be quantified using a single numerical score. This study demonstrated that it is possible to quantify the collagen's irregularity in an objective manner, and that the study of the fractal properties of the collagen shapes is likely to reveal more about its structure and the complex behaviour of its development. PMID- 12768880 TI - [Aneurysmal bone cysts of the nasal cavity. Description of a case and review of the literature]. AB - INTRODUCTION: Aneurysmal bone cyst (ABC) are benign tumor-like lesions of the bone that occur at any age with a slight female predominance. The most common sites are long bones and vertebrae bodies. MATERIALS AND METHODS: We report a case of 54 year-old man with nasal respiratory difficulty and a previous history of nasal fracture. RMN and TC-scan showed a voluminous nasal lesion, which was surgically removed. Tissue sample was formalin fixed (10%) and routinely processed to paraffin after decalcifying treatment (Osteodec, Bioptica). Section were stained with haematoxilin-eosin. RESULTS: At histological level the lesion was characterized by multilocular cyst-like spaces filled with erythrocytes separated by connective septa lacking the smooth muscle walls and endothelial lining of normal vessels. Histological features are consistent with a diagnosis of ABC. DISCUSSION: We report a case of ABC of nasal cavity. The previous history of nasal fracture and the fact that pre-existing or concurrent were not found lead us to suppose a relationship between the fracture and ABC. Teleangiectatic osteosarcoma, fibrous dysplasia and giant-cell tumor were excluded according to clinical and morphological features. PMID- 12768881 TI - [Primary leiomyoma of the lung, with clear-cell features and admixed alveolar spaces: description of a case presenting with pneumothorax]. AB - OBJECTIVE: To report about a primary leiomyoma of the lung, with clear cell features and with admixed alveolar epithelium. RESULTS AND CONCLUSIONS: The patient, a 18-year-old female, non smoker, presented with a well-delimited pleuro pulmonary mass, 6,5 cm across, located in the right upper lobe. The mass was discovered 5 years previously, following an episode of spontaneous pneumothorax; initially interpreted as an intralobar pulmonary sequestration, it was subsequently removed due to a small dimensional increase. A gynaecological examination, a thoracic CT-scan obtained 2 years after operation, and a recent abdomino-pelvic echography were negative. The patient is alive and well 7 years after surgery. Microscopically, the lesion was well delimited and it was punctuated by multiple cysts of variable dimensions, covered by bland cubic cells. The solid part of the lesion was composed by oval, clear cells, merging with fusiform elements with densely eosinophilic cytoplasm. Significant atypia, mitoses and necrosis were absent. Immunohistochemically, both oval and spindle cells were strongly positive for vimentin, smooth muscle actin, desmin and h caldesmon, negative for cytokeratin, EMA, CD-34, CD-10, CD-117, HMB-45, TTF-1, BCL-2, GFAP, calretinin, chromogranin, estrogen and progesterone receptors. Cells covering the cystic spaces were positive for cytokeratin, TTF-1 and EMA, negative for all the other antibodies tested. Smooth muscle proliferations of the lung are briefly discussed, together with the most important problems in differential diagnosis. PMID- 12768882 TI - [Adenoid cystic carcinoma of the anterior segment of the tongue]. AB - The authors describe a rare case of adenoid-cystic carcinoma localized in the anterior tongue. The neoplasia is made up of epithelial and myo-epithelial cells, organized frequently in islands with cribriform aspect, was circumscribed by a thin fibrous capsule that was focally infiltrated in the proximity of the deep margin. At the physical examination no local regional lymph nodes were absent. The surgical excision and radiotherapy constituted the only therapeutic protocol. Absence of local relapse and/or distant metastasis after eight years from the operation confirm the low aggressiveness of the neoplasia when localized in the anterior part of the tongue. PMID- 12768883 TI - [Renal capsuloma: description of a case with predominantly muscular differentiation]. AB - The Authors report a case of Renal Capsuloma with a histologic biphasic pattern constituted of epithelioid and spindle cells. The lesion is observed in a nephrectomy specimen as incidental finding concurrent with a Papillary Transitional Cell Carcinoma of calices and renal pelvis. The lesion presents a distinct muscular immunophenotype (actin+, desmin+), with coexpression of the melanogenesis marker (HMB45+) in a lower cellular population. This combined immunophenotype is a characteristic feature of the Perivascular Epithelioid Cell's (PEC) lesions. Actually Renal Capsuloma is a PEC's lesion (Pecoma) closely related to Angiomyolipoma (AML). Particularly Capsuloma is thought as a variant of AML predominantly composed by muscular spindle cells (monotypic spindle cell leiomyoma like variant of AML). In the presented case the spindle cell component is associated with an epithelioid cellular population which also shows a distinct muscular differentiation. The Authors have not found in the literature the association between Renal Capsuloma and Papillary Transitional Cell Carcinoma of the renal pelvis. PMID- 12768884 TI - [Technique for macroscopic identification of lymph nodes for malignant neoplasia staging]. PMID- 12768885 TI - [Implant survival and success rates in partially edentulous patients--Part I]. AB - The use of osseointegrated implants is an accepted procedure for the treatment of complete or partially edentulous jaws. The purpose of this prospective long-term study was to evaluate the survival and success rates of endosseous dental implants (machined surface) in partially edentulous patients. A total of 76 patients (47 females, 29 males) with 214 implants (68 maxillary, 146 mandibulary) were evaluated. Their mean age was 45, ranging from 18 to 76 years. The implant length varied between 7 and 15 mm, the diameter between 3.4 and 6 mm. Nine implants were lost before abutment connection. Follow-up visits were scheduled at 3 and 6 months, 1 year, 2 and 3 years after insertion of the prostheses. Radiographic examination included intraoral periapical films to evaluate the horizontal and vertical bone loss. During the follow-up period, 18 implants in 10 patients were classified as failures. Fifteen implants were lost, and three implants were considered failures because of vertical bone loss greater than 0.2 mm per year after the first year of functional loading. The marginal bone loss averaged 2.1 mm in the maxillae and mandibles. According to the Kaplan-Meier estimator the probable implant survival and success is at 93% and 88% respectively after 63 months. Long implants (> 10 mm) have a significantly better chance of survival than shorter implants (< or = 10 mm). The results of this prospective study indicate that 3i endosseous dental implants provide an average foundation for implant-supported restorations. PMID- 12768886 TI - [Success rates of implant supported prostheses in partially edentulous patients- Part II]. AB - The use of dental implants is an established procedure for the treatment of partially edentulous patients. However, there is only little knowledge about the prosthetic outcome of implant-supported restorations. The purpose of this prospective long-term study was to evaluate success rates of implant-supported restorations in partially edentulous patients three years after loading. The study group consisted of 76 patients (47 females, 29 males) with 214 3i-implants (machined surface). 64 implants were used to support single crowns, 81 implants for 81 splinted crowns (36 units), 17 implants for seven bridges and 61 implants for 23 cantilevered bridges. For prosthesis construction, UCLA-Abutments were used. Screw connection was achieved by gold-alloy-coated square head screws and titanium hexagon head screws. Clinical follow-up visits were performed at the time of prosthesis insertion, three months, six months, one year, two years and three years after insertion of the prosthesis. The evaluation of the superstructures was performed according to the success criteria defined by Walton (1998). The success rate for the prosthetic reconstructions was estimated at 84.9% 36 months after loading. The success rate was 88.6% for fixed partial dentures, 86.4% for single crowns and 76% for cantilevered bridges. Technical problems included screw loosening, screw fractures, framework fractures, veneering material failures and failures because of implant loss and following remake or modification of the superstructures. Since mainly technical complications such as screw loosening and screw fractures were causing problems, the connection between implant and superstructure should be improved. PMID- 12768887 TI - Surgical removal of a dislocated lower third molar from the condyle region: case report. AB - INTRODUCTION: Migration of dislocated lower third molar into the condylar region is quite rare. Attention should be taken to avoid condyle fracture. METHODS: 49 year-old patient had an ectopic lower left third molal in the condylar region, suffered from a submandibular and masseter space abscess. Removal of the molar via intraoral approach was chosen avoiding facial nerve branches and unnecessary scar formation. Coronoid process is removed, the tooth is separated and removed. The defect is filled with iliac cancellous bone. The coronoid process was fixed as a cover with a resorbable plate and screws (BIONX). RESULTS: Removal via intraoral approach is possible. Hypesthesia existed postoperatively, became normal later. CONCLUSION: Annual observation is strongly recommended. Intraoral approach is superior to the extraoral approach. Removal of the coronoid process minimizes the masticator forces. Separation of the tooth is essential. Filling the defect with cancellous bone accelerates the healing. PMID- 12768888 TI - [An in vitro model for the simulation of approximal wear]. AB - Several methods have been developed to evaluate attritional wear of restorative materials. A distinction was made between attritional wear in occlusal contact areas (OCA) and wear in contact-free occlusal areas (CFOA). Only limited data dealing with approximal wear in vitro are available. Using a new configuration of the computer-controlled masticator, approximal wear of two composites (P-50, 3M and Tetric Ceram, Ivoclar-Vivadent) was assessed in a two-body wear test after thermo-mechanical loading. Wear was quantified using a profilometer. In addition, micromorphology was assessed by SEM. Wear showed a non-linear pattern, which was comparable to occlusal abrasion. After the first loading cycle, wear increased significantly (p < or = 0.05), and subsequently decreased. After a 5-year equivalent, the mean substance loss for composite specimens was 20.3 +/- 15.6 microns for P-50 und 17.5 +/- 3.1 microns for Tetric Ceram. Approximal wear between enamel surfaces was 3.9 +/- 4.3 microns. In another experiment, the influence of the approximal wear on an enamel bonding agent was assessed. After thermo-mechanical loading, specimens were immersed in a demineralisation gel. The protective potential of the sealant was assessed under a polarized light microscope. The results suggested that partial sealant loss could lead to caries like demineralisation in the affected areas. PMID- 12768889 TI - WHO Expert Committee on Specifications for Pharmaceutical Preparations. AB - This report presents the recommendations of an international group of experts convened by the World Health Organization to consider matters concerning the quality assurance of pharmaceuticals and specifications for drug substances and dosage forms. Of particular relevance to drug regulatory authorities and pharmaceutical manufacturers, the report discusses activities related to the development of The International Pharmacopoeia and basic tests for pharmaceutical substances and dosage forms, as well as quality control of reference materials, good manufacturing practices (GMP), stability studies, inspection, hazard analysis, procurement, storage and other aspects of quality assurance of pharmaceuticals, and regulatory issues. The report is complemented by a number of annexes, including recommendations on the risk of transmitting animal spongiform encephalopathy agents via medicinal products, guidelines on GMP for pharmaceutical products, a model certificate for GMP and guidance on a GMP inspection report. The final annexes provide guidance on the application of Hazard Analysis and Critical Control Point (HACCP) method to pharmaceuticals, good storage practices and a procedure for assessing acceptability of pharmaceutical products for purchase by United Nations agencies. PMID- 12768890 TI - Diet, nutrition and the prevention of chronic diseases. AB - Shifting dietary patterns, a decline in energy expenditure associated with a sedentary lifestyle, an ageing population--together with tobacco use and alcohol consumption--are major risk factors for noncommunicable diseases and pose an increasing challenge to public health. This report of a Joint WHO/FAO Expert Consultation reviews the evidence on the effects of diet and nutrition on chronic diseases and makes recommendations for public health policies and strategies that encompass societal, behavioural and ecological dimensions. Although the primary aim of the Consultation was to set targets related to diet and nutrition, the importance of physical activity was also emphasized. The Consultation considered diet in the context of the macro-economic implications of public health recommendations on agriculture and the global supply and demand for fresh and processed foodstuffs. In setting out ways to decrease the burden of chronic diseases such as obesity, type 2 diabetes, cardiovascular diseases (including hypertension and stroke), cancer, dental diseases and osteoporosis, this report proposes that nutrition should be placed at the forefront of public health policies and programmes. This report will be of interest to policy-makers and public health professionals alike, in a wide range of disciplines including nutrition, general medicine and gerontology. It shows how, at the population level, diet and exercise throughout the life course can reduce the threat of a global epidemic of chronic diseases. PMID- 12768891 TI - [Internet in medicine--development and perspectives]. AB - Internet is one of information technologies marking the transition from the second to the third millennium. The present role and expansion of Internet in medicine and healthcare is reviewed together with the perspective of further development. The beginning and initial expansion of the use of Internet in medicine are described. The World Wide Web (WWW or Web) is recognized as a major reason for this expansion, reaching a state described as a Web-pandemic. The rapid increase of the number of papers dealing with Internet in medical literature is presented as well as the appearance of several journals dedicated to Internet in medicine. First specialized symposia, among them MEDNET world conferences, are noted. First uses of Internet in medicine comprised databases, discussion groups, electronic newsletters, software archives and online public access catalogues. The appearance of the Web has led to a significant improvement of the Internet use in medicine, which is reflected in an exponential increase in the number of publications. It is noted that Internet allows "to do old things in new ways", but also "to do new things". It has become clear that the information revolution evoked by the internet shall leave a deep trace in medicine, as health information has become accessible to the public and ceased to be in exclusive control of health professionals. New medical fields--telemedicine and cybermedicine--appeared as the result of the development and global expansion of information and communication technologies, with cybermedicine dealing more specifically with the use of Internet. The advantages and disadvantages of cybermedicine are discussed, and major problems related to the quality of health information are highlighted. Several systems for quality criteria of health related Web-sites are described, indicating that Websites have to conform with the quality criteria such as transparency and honesty, accountability, privacy and data protection, currency, accountability and accessibility. A review of the development of the use of Internet in medicine in Croatia is given. It is ever more recognized worldwide that the Internet is capable to create conditions for partnership between patients and clinicians. In concluding remarks, the reasons are given for a sooner access to the Internet and its daily use in healthcare activity. PMID- 12768892 TI - [What the citizens of Croatia think about their health]. AB - AIM: Since self-perceived health represents an important indicator of health status, the aim of this study was to explore socioeconomic differences in self perceived health in Croatia. METHODS: The study was conducted during the 1997 1999 period in fourteen Croatian counties. A sample of 5,048 respondents aged 18 years and older were recruited by multistaged stratified sampling. Data were collected using an anonymous questionnaire which consisted of questions on the respondents' socioeconomic characteristics and self-perceived health. RESULTS: While 20.7% of the respondents evaluated their health as poor, 25.9% evaluated it as excellent or very good. Women, older respondents and those with lower educational level and lower income more frequently described their health as poor. When comparing their health to the health of age-matched persons, 23.6% of the respondents thought their health was better and 21.0% thought their health was worse than the health of persons of the same age. Women, older respondents and those with lower educational level and lower income more frequently thought their health was worse than the health of their coevals. As many as 53.7% of the respondents thought their health had not changed compared to the previous year, and 31.7% thought their health was worse than in the previous year. Older respondents, respondents with lower income and those with lower educational level more frequently thought that their health had worsened compared to the previous year. DISCUSSION: Results of the study revealed a complex relationship between socioeconomic factors and self-perceived health. Men and women differed significantly in self-perceived health. Women mostly described their health as being poor or satisfactory, while men described it as excellent or very good. These results, similar to those in a large body of literature, raise the question of gender differences in the perception of symptoms. In our research, self perceived health was poorer in older respondents. This also applies to the comparison of self-perceived health to the health of age-matched persons. An exception was the group of respondents aged > 76; their self-perceived health was described as either much worse or much better than the health of their coevals. Older persons are inevitably sicker than younger ones and poor self-perceived health would be expected. Nevertheless, the relation between self-perceived health and age is sometimes ambiguous in the literature. Income and education are strong markers of socioeconomic status. In our research, respondents with lower income as well as those with lower educational level had poorer self-perceived health. Education is a particularly strong marker of socioeconomic status, and it predicts self-perceived health better than occupation does. A study of self perceived health does not always give an accurate insight into the real self perceived health of the respondents. For example, if a person, otherwise in good health, suffered from an acute condition, he could describe his health as being poor at that moment. We tried to minimize this potential bias by adding two questions in self-perceived health assessment--"How do you perceive your health compared to your health a year ago?" and "How do you perceive your health compared to health of the persons of the same age?" Women, older respondents, respondents with lower educational level and lower income generally described their health as poor more often. Since self-perceived health is correlated with health care utilization, these results could be used to identify the specific population groups who are more likely to be potential users of health care services. CONCLUSION: Since education and income represent socioeconomic parameters, it could be concluded that self-perceived health of the respondents with lower socioeconomic status is poorer than self-perceived health of those with higher socioeconomic status. PMID- 12768893 TI - [Study of friction and loosening in hip endoprostheses]. AB - AIM: Like any other operative procedure, the implantation of hip prosthesis is associated with certain complications, which diminishes the value and purpose of such a procedure. One of the complications in artificial hip implantation is loosening of the alloplastic material. Therefore, the aim of this study was to examine the effect of lubrication on the torsional moment and its role in the loosening of the femoral component, using an experimental mechanical model. The following hypothesis was tested: the magnitude of torsional loading in the "bone endoprosthesis-bone cement system" is similar to any other known loading. METHODS: The testing device was constructed with the possibility of simulation of positions similar to original performances in the implanted hip prosthesis. It refers primarily to the possibilities of achieving definite forces and velocities. The intention was to point quantitatively to the role of friction moment between the acetabular and femoral endoprosthesis part. Trials were conducted by combining 7 types of loading and 4 kinds of lubrication: dry, water, plasma, and light oil. The testing joint (Ring's prosthesis) was connected through tensometric measuring shaft upon the working forepart oscillating mechanism. Graded by the changeable static loading by means of the pendulum and via lever mechanism the testing joint was loaded by force from 610 to 7137 N. As the cause of friction resistance in the moving joint, torque deformaties of the measuring shaft occurred. The testing joint enabled oscillating movement using a four-part mechanism. In this way, it was possible to define not only the maximum values of the frictional moment (or the coefficient of friction) during one movement cycle but also to examine its relation to the kind of lubrication. Change in the measuring torsional moment were computer recorded. Before each trial, the gauging of the complete outfit was performed. Thereafter, cleaning of the frictional surfaces of the whole outfit was done. RESULTS: The results obtained in combination with lubrication showed a slight increase in the values of the frictional moment. With dry lubrication and greater loading, an extremely progressive gradient of change was recorded. The course of change in the coefficient of friction was essentially different from the course of change in the frictional moment. It was noted particularly during trials with lubrication. In trials without lubrication, a constant increase of loading (force) resulted in a progressive increase in the coefficient of friction, similar to the friction moment. Such a character of the friction moment increase in the observed loading field was explained by the presence of boundary friction in cases with lubrication and by dry friction in cases without lubrication. In dry friction, scratching occurs relatively early, at a loading of F = 1854 N. It occurs with substances of approximately the same hardness like Ring's prosthesis, where the acetabular and femoral prosthesis parts are of metal characteristics. CONCLUSION: The increase in the frictional moment within the observed loading range can be explained by the presence of bordering friction in cases with lubrication, and of dry friction in cases without lubrication. Contrary to this, dry friction relatively early leads to "scratching", especially when sparing materials of similar hardness are combined. PMID- 12768894 TI - [Effect of viscoelasticity on corneal endothelial cell loss in phacoemulsification]. AB - One of the most important factors for successful intraocular surgery is deep anterior chamber which is maintained using viscoelastics that protect corneal endothelial cells. This single endothelial cell layer may sustain a varying degree of injury during surgery and is known to be incapable of regeneration. THE AIM: Of the study was to determine optimal visoelastics for the protection of corneal endothelial cells during phacoemulsification and intraocular lens implantation. PATIENTS AND METHODS: A series of 156 patients were included in this prospective study. Patients were randomly assigned to three groups based on the type of viscoelastics used. In the first group, 1.4% sodium hyaluronate, in the second group a combination of 4% sodium chondroitin sulfate and 3% sodium hyaluronate, and in the third group 3% hyaluronate was used. Endothelial cell count and morphology changes were determined centrally with a specular microscope on the day before, on day 7 and one month after surgery. Intraocular pressure was measured before surgery, and at 6 hours, 24 hours, 7 days and 30 days of the surgery. Pachymetry as a measure of "endothelial pump" function was evaluated before surgery and 7 and 30 days postoperatively. All surgeries were performed by two surgeons using the same technique. RESULTS: Endothelial cell loss was lowest (16.1%) in the eyes protected with a combination of 4% sodium chondroitin sulfate and 3% sodium hyaluronate. In the groups where 1.4% sodium hyaluronate and 3% hylunorate were used, the loss was 7.6%; and 7.9%, respectively. Although endothelial cell loss was not significantly different among the three groups, the lowest morphology change rate recorded in group 2 suggested that a combination of chondroitin and hyaluronate offered the best endothelial protection. There was no statistically significant difference in the postoperative intraocular pressure increase and pachymetry values among the study groups. PMID- 12768895 TI - [Renal biopsy in children with isolated microhematuria]. AB - Isolated microscopic hematuria (IMH) in children always raises the question whether, besides other examinations, there is a need of performing a renal biopsy. Many authors consider IMH to be a minor abnormality where pathologic glomerular changes are not likely to be found, however, general agreement has not yet been achieved. The aim of the study was to evaluate the contribution of renal biopsy to the diagnosis of the disease in IMH. PATIENTS AND METHODS: Renal biopsy was performed in 54 children with IMH (22 boys and 32 girls, mean age 8.2 and 8.5 years, respectively) in whom urologic abnormalities, hypercalciuria, systemic diseases, coagulopathy or overt family history of renal disease were excluded. The mean duration of IMH prior to biopsy was 2.8 years. Biopsy specimens were examined by light (LM), immunofluorescent (IF) and electron microscopy (EM). RESULTS: Glomerular abnormalities were found in 43 (79.6%) patients. On LM 18 patients had normal glomeruli (NG), 22 mesangial proliferative glomerulonephritis (MEPGN), 9 focal glomerulosclerosis (FGS), 3 focal glomerulonephritis (FGN) and 2 membranoproliferative glomerulonephritis (MPGN). IF revealed 2 cases of NG, 5 cases of MEPGN, and all 3 cases of FGN as IgA nephropathy. EM detected GBM changes consistent with Alport syndrome in 21 patients, 7 of them with NG, 9 with MEPGN and 5 with FGS on LM. Diffuse thinning of GBM was found in 10 children, 7 with NG and 3 with MEPGN on LM. In 5 cases subepithelial hump-like deposits, which were considered to be the sign of acute postinfectious glomerulonephritis in resolution, were found. One of 2 cases of MPGN showed to be type II (DDD). On follow-up, 6 of 21 children with changes consistent with Alport syndrome developed clinical signs of the syndrome. Further surveillance is needed to confirm the significance of EM findings in others. CONCLUSION: The authors concluded that in children with IMH renal biopsy is justified and should always be analyzed by light, immunofluorescent and electron microscopy. PMID- 12768896 TI - [Cystic cystitis in children]. AB - Cystic cystitis is a common urinary bladder disease with a histologic picture of nodular lesions of the bladder mucosa. Intralesional lymphoid proliferation due to recurrent urinary tract infection play a major role in the occurrence of cystic cystitis. It is often found together with vesicoureteral reflux and other urinary tract anomalies and urodynamic disturbances. Examination of 116 children with cystic cystitis revealed it to be associated with urinary tract anomalies, especially vesicoureteral reflux. The average age at diagnosis is 7 years. The disease often needs longterm prophylaxis for urinary tract infections, and has favorable prognosis. PMID- 12768897 TI - [Ultrasonography in acute pelvic pain]. AB - Acute pelvic pain may be the manifestation of various gynecologic and non gynecologic disorders from less alarming rupture of the follicular cyst to life threatening conditions such as rupture of ectopic pregnancy or perforation of inflamed appendix. In order to construct an algorithm for differential diagnosis we divide acute pelvic pain into gynecologic and non-gynecologic etiology, which is than subdivided into gastrointestinal and urinary causes. Appendicitis is the most common surgical emergency and should always be considered in differential diagnosis if appendix has not been removed. Apart of clinical examination and laboratory tests, an ultrasound examination is sensitive up to 90% and specific up to 95% if graded compression technique is used. Still it is user-depended and requires considerable experience in order to perform it reliably. Meckel's diverticulitis, acute terminal ileitis, mesenteric lymphadenitis and functional bowel disease are conditions that should be differentiated from other causes of low abdominal pain by clinical presentation, laboratory and imaging tests. Dilatation of renal pelvis and ureter are typical signs of obstructive uropathy and may be efficiently detected by ultrasound. Additional thinning of renal parenchyma suggests long-term obstructive uropathy. Ruptured ectopic pregnancy, salpingitis and hemorrhagic ovarian cysts are three most commonly diagnosed gynecologic conditions presenting as an acute abdomen. Degenerating leiomyomas and adnexal torsion occur less frequently. For better systematization, gynecologic causes of acute pelvic pain could be divided into conditions with negative pregnancy test and conditions with positive pregnancy test. Pelvic inflammatory disease may be ultrasonically presented with numerous signs such as thickening of the tubal wall, incomplete septa within the dilated tube, demonstration of hyperechoic mural nodules, free fluid in the "cul-de-sac" etc. Color Doppler ultrasound contributes to more accurate diagnosis of this entity since it enables differentiation between acute and chronic stages based on analysis of the vascular resistance. Hemorrhagic ovarian cysts may be presented by variety of ultrasound findings since intracystic echoes depend upon the quality and quantity of the blood clots. Color Doppler investigation demonstrates moderate to low vascular resistance typical of luteal flow. Leiomyomas undergoing degenerative changes are another cause of acute pelvic pain commonly present in patients of reproductive age. Color flow detects regularly separated vessels at the periphery of the leiomyoma, which exhibit moderate vascular resistance. Although the classic symptom of endometriosis is chronic pelvic pain, in some patients acute pelvic pain does occur. Most of these patients demonstrate an endometrioma or "chocolate" cyst containing diffuse carpet-like echoes. Sometimes, solid components may indicate even ovarian malignancy, but if color Doppler ultrasound is applied it is less likely to obtain false positive results. One should be aware that pericystic and/or hillar type of ovarian endometrioma vascularization facilitate correct recognition of this entity. Pelvic congestion syndrome is another condition that can cause an attack of acute pelvic pain. It is usually consequence of dilatation of venous plexuses, arteries or both systems. By switching color Doppler gynecologist can differentiate pelvic congestion syndrome from multilocular cysts, pelvic inflammatory disease or adenomyosis. Ovarian vein thrombosis is a potentially fatal disorder occurring most often in the early postpartal period. Hypercoagulability, infection and stasis are main etiologic factors, and transvaginal color Doppler ultrasound is an excellent diagnostic tool to diagnose it. Acute pelvic pain may occur even in normal intrauterine pregnancy. This may be explained by hormonal changes, rapid growth of the uterus and increased blood flow. Ultrasound is mandatory for distinguishing normal intrauterine pregnancy from threatened or spontaneous abortion, ectopic pregnancy and other complications that may occur in patients with positive pregnancy test. Incomplete abortion is visualized as thickened and irregular endometrial echo with certain amount of intracavitary fluid. If applied, color Doppler ultrasound reveals low vascular resistance signals in richly perfused intracavitary area. Transvaginal sonography has high sensitivity and specificity in visualization of uterine and adnexal signs of ectopic pregnancy. Color Doppler examination may aid in detection of the peritrophoblastic flow. Furthermore, it facilitates detection of ectopic living embryo, tubal ring or unspecific adnexal tumor. Corpus luteum cysts and leiomyomas are another cause of pelvic pain during pregnancy, which can be correctly diagnosed by ultrasound. Detection of uterine dehiscence and rupture in patients with history of prior surgical intervention on uterine wall relies exclusively on correct ultrasound diagnosis. In patients with placental abruption sonographer detects hypoechoic complex representing either retroplacental hematoma, subchorionic hematoma or subamniotic hemorrhage. In closing, ultrasound has already become important and easily available tool which can efficiently recognize patients with possibly threatening conditions of different origins. PMID- 12768898 TI - [Primary pulmonary hypertension in pregnancy: case report]. AB - Primary pulmonary hypertension (PPH) is a rare, progressive and currently incurable disease characterised by an increase in pulmonary artery pressure without a demonstrable cause. When associated with pregnancy, the maternal mortality ranges from 30 to 50%. In this report we present a 26-year old patient diagnosed with primary pulmonary hypertension during her twenty-first week of gestation. She was treated with a low-molecular-weight heparin and sedatives and underwent pregnancy termination without complications. Although cases have been reported with positive maternal and fetal outcome, the available evidence suggests that pregnancy with primary pulmonary hypertension should be avoided. Thus, in cases of pregnancy occurring, a therapeutic abortion is indicated. PMID- 12768899 TI - [Sterile peritonitis after administration of icodextrin]. AB - INTRODUCTION: Icodextrin is an 7.5% isoosmotic solution of the glucose polymer maltodextrin, recently frequently used in continuous ambulatory peritoneal dialysis (CAPD). It improves ultrafiltration and decreases glucose absorption. Among side effects of icodextrin, skin reactions and sterile peritonitis have been described. The authors present two cases of sterile peritonitis after icodextrin. CLINICAL PRESENTATION: In these two patients, the adverse events included peritoneal reaction to icodextrin solution. The reaction did not occur immediately after the initial administration of icodextrin, but after 70 (case 1) and 412 days (case 2) of regular use. This reaction to icodextrin solution resembled chemical peritonitis, however, the clinical picture was not absolutely typical of bacterial peritonitis either. Both patients had cloudy dialysate, elevated WBC in dialysate, no microorganisms were isolated, and no signs of allergic reaction could be detected. The clinical course was characterized by rapid remission upon isodextrin withdrawal. One patient received empiric antibiotic therapy, the other did not. Upon switching from icodextrin to glucose based dialysate, the two patients were not challenged to icodextrin, and were doing well, without any symptoms of peritonitis. DISCUSSION AND CONCLUSION: The icodextrin solution contains an isoosmolar glucose polymer which allows longterm stable ultrafiltration. Among side effects, cutaneous reactions have been described (exfoliative, in the form of vesicles, psoriatic plaque or generalized exanthema in the form of pustules), which may occur in as many as 15% of patients. These symptoms and signs disappear upon discontinuation of icodextrin. Icodextrin side effects also include mild abdominal pain, allergic and hypersensitivity reactions, and recently sterile peritonitis. The patients described developed a clinical picture suggestive of chemical peritonitis. From the effluent no microorganisms were isolated, and the clinical picture was not absolutely typical of bacterial peritonitis either. Discontinuation of icodextrin was followed by immediate clearing of the effluents and normalization of WBC count. PMID- 12768900 TI - [Views on health, disease and human life]. AB - We should not conceptualize health as a normal biological state. The WHO definition, equating health with physical, psychological, and social satisfaction is not adequate. Health should be defined as a life situation unaffected by pain, injuries, physical, and/or psychological dysfunctions. Most of the time we are only partially healthy, just as we are only partially free (J.J. Rousseau). Both health and illness should be comprehended as normal biological states, which are constantly interchanging. Individual variations are the consequences of a complex interaction of genetic and environmental factors. These variations are the source of our individuality, talents, and specific skills. Genetic mutations, as is well known, can lead to a range of diseases, which modern molecular medicine is trying to prevent or treat with success. The sense of pain is an important defense mechanism that has evolved in many species, including ours. It is an alarm system indicating problems and pathogenic developments. The sense of pain is of huge importance in maintaining health. There is yet another important phenomenon in human life--the addiction syndrome. As well researched, addiction is related to specific brain receptors. In some individuals these receptors are extremely sensitive to psychoactive substances, such as alcohol, nicotine, and recrational drugs, leading to addiction. In contemporary societies, the symptoms of addiction can also be related to the more extreme forms of professional and sports ambitions. Contemporary medical science has succeeded in prolonging human life. The costs, however, have also risen due to the aging population. This fact is not only an economic or a social, but also a political problem. Human aging should not be viewed only as a process of wearing the biological machine off, but primarily as a gradual loss of genetic control over life functions. PMID- 12768901 TI - [Intrathecal baclofen in spasticity]. PMID- 12768902 TI - [Intrathecal baclofen in the treatment of severe spastic tetraplegia and dystonia in children and adolescents]. AB - INTRODUCTION: Continuous intrathecal baclofen has been used over the past years especially in adult patients with spasticity of spinal origin. Children and young adults with severe spasticity and dystonia of cerebral origin are difficult to treat in spite of optimal systemic antispasmotic therapy with baclofen, tizanidine, dantrolene and/or diazepam. Intrathecal baclofen has therefore been applied in a group of young patients. MATERIAL AND METHODS: Eight children and young adults from East Denmark with spasticity and 12 with dystonia aged 3-18 years (median 10.9 years) were tested, operated and treated with continuous intrathecal baclofen for a period of 2-64 months (median 22.2 months). Registration of efficacy, fillings, adjustments of baclofen and other therapies were performed in an out patient setting since 1995. RESULTS: Spasticity in lower extremities was reduced from Ashworth score 3.5-4.5 (median 4.2) to Ashworth score 2.5-4.0 (median 2.9; p < 0.001) during infusion with baclofen 5-33 micrograms/kg/24 hours (median 19 micrograms/kg/24 hours). The infusion catheter tip was placed at levels Th1-Th12 (median Th7.5). Peroral baclofen was reduced from an average of 5.0 to 0.44 mg/kg/24 hours, tizanidine from 0.4 to 0.1 mg/kg/24 hours, and dantrolene from 4.0 to 0.4 mg/kg/24 hours. After initial adjustments successively increased dosages of average 0.46 microgram/kg/month were needed to maintain the same level of efficacy. In questionnaires parents or guardians rated less spasticity in lower extremities in 15 out of 19 patients, and less pain in 13 out of 19 patients. CONCLUSION: Continuous intrathecal baclofen was effective in treating severe spasticity and dystonia of cerebral origin with major effect on muscles of the lower extremities, pelvis, and back and in particular opisthotonus was relieved. Efficacy on upper extremities was far less pronounced. PMID- 12768903 TI - [Treatment of gastroduodenal obstruction with self-expanding stent]. AB - Self-expanding metal stents are used in some centres as non-surgical palliation for malignant gastric outlet obstruction as an alternative to surgery. No randomized studies exist comparing these two treatment modalities. Descriptive series have shown high success rates of 82-100% for stent placement, and 75-100% of the patients experience relief of the vomiting and ability to eat semisolid to solid food. Reobstruction was seen in about 15% of the patients requiring a second stenting. Serious complications to the stenting procedure have been reported in case studies. It is recommended to perform randomized studies of stenting versus open and laparoscopic surgical bypass for malignant gastric outlet obstruction. PMID- 12768904 TI - [Malignant hyperthermia]. AB - Malignant hyperthermia susceptibility is a rare pharmacogenetic muscular disorder that may lead to potentially fatal complications in routine anesthesia. Malignant hyperthermia is triggered by volatile anesthetics and succinylcholine. In this article the pathophysiology of malignant hyperthermia, its signs, symptoms and treatment as well as safe anesthetic agents for individuals susceptible to malignant hyperthermia are discussed. Although progress is made in our knowledge of the genetics behind malignant hyperthermia, muscle biopsy and in-vitro contracture testing is still the only way to diagnose malignant hyperthermia susceptibility in the vast majority of cases. Danish Malignant Hyperthermia Center at the Anesthetic Department of Herlev University Hospital, DK-2730 Herlev, takes care of all testing for malignant hyperthermia in Denmark and keeps a registry of all Danes with known or suspected susceptibility to malignant hyperthermia. PMID- 12768905 TI - [Webbing of the neck and nasal bone--prenatal screening for Down syndrome]. AB - The risk of giving birth to a child with Down syndrome increases with maternal age. If in Denmark the mother is older than 35-years she belongs to a high-risk group and will be offered amniocentesis or chorionic villus sampling. We describe different scenarios of risk calculation for Down syndrome based on combinations of nuchal translucency, nasal bone, and/or integrated hormone test. The combination of nuchal translucency and visualisation of the nasal bone between gestational age 11 weeks and 13 + 6 has turned out to be as good a predictive marker as nuchal translucency and integrated hormone test. PMID- 12768906 TI - [Monitoring of irrigating fluid absorption in transvesical resection of the myometrium]. AB - INTRODUCTION: Monitoring of the absorption of irrigating fluid during transcervical resection of the myometrium was studied. Irrigating fluid under pressure was used. Absorption can give symptoms as seen in transurethral resection syndrome. MATERIAL AND METHODS: A method using ethanol labelled irrigation fluid measuring ethanol in expired air was used. Serum natrium and absorbed irrigation fluid estimated by weight was measured. A total of 100 women were included. The investigation was part of the daily routine with varying anaesthetists and nurses. RESULTS: Four patients with an absorption of more than 1200 ml were identified. The end-tidal ethanol and serum natrium deficit were equally accurate in determination of this. In absorption of less than 1200 ml, the serum natrium deficit and end-tidal ethanol were not correlated to absorbed irrigation fluid judged by weighing. CONCLUSION: Determination of the end-tidal ethanol during transcervical resection is recommended as a supplement. The method is non-invasive and simple and can be repeated at any time without interruption of surgery. PMID- 12768908 TI - [Scapulae alatae--angels' wings. A study of 64 patients treated with braces and physical therapy at the Viberg's hospital]. AB - INTRODUCTION: Scapula winging is characterized by a paresis/paralysis of either m. serratus anterior innervated by n. thoracis longus or m. trapezius innervated by n. accesorius. This investigation reveals the results after bracing and muscle training in patients suffering from scapula winging. MATERIAL AND METHODS: A prospective, consecutive series consisting of 64 patients, 43 males and 21 females with a mean age of 37.8 years (15-75 years), were treated with bracing and muscle training in 1998-2001. Prior to the treatment of bracing all patients suffered from shoulder pain, shoulder fatigue and limited range of motion as well as cosmetic complaints, 28 patients had shoulder pain. RESULTS: At the time of the investigation all 64 patients had stopped using the brace after an average of 11 months (3-29 months). A total of 61 patients had returned to their previous work and sports activities. By the time of the one year follow-up control 18 (50%) patients had gained normal shoulder functions and had no complaints and no visible scapula winging. Ten (32%) had minor disability and two (6%) had major disability. Two patients had relapse of their scapula winging. (Both patients had formerly had normal ENG/EMG). DISCUSSION: We find that the combination of bracing and muscle training is successful in treating scapula winging. Electroneurography (ENG)/electromyography (EMG) was routinely performed but did not show any prognostic value for the treatment in our investigation. PMID- 12768907 TI - [Remifentanil and eye surgery. A randomized, clinical comparison of propofol/remifentanil anesthesia and propofol/fentanyl/alfentanil anesthesia]. AB - INTRODUCTION: The aim of this study was to investigate how many patients, after anesthesia with either propofol/remifentanil or propofol/fentanyl/alfentanil, within 20 minutes from the end of surgery could be transferred directly to the general ward. The number of undesired preoperative incidents, the anesthetists', the surgeons', and the patients' evaluations of the anesthesia were registered. An evaluation of the economic consequences of the two methods was also intended. MATERIAL AND METHODS: The study was clinically controlled, randomised, and partly blinded. A total of 80 patients undergoing eye surgery were recruited. The patients were scored 10, 15, and 20 minutes after the end of surgery according to a modified Aldrete score. With sufficient awakening score, the patients were transferred to the general ward. RESULTS: Thirty-six patients in each group underwent the examination. In the propofol/remifentanil-group 31 (86%) could be transferred to the general ward compared to 15 (42%) in the proponol/fentanyl/alfentanil-group. In the propofol/remifentanil-group there were less reactions to the start of surgery, more episodes with preoperative hypotension and postoperative shivering. Otherwise there were no differences between the groups. It was estimated that the additional expenses for medcine were by far outweighed by the lower costs postoperatively. DISCUSSION: With a propofol/remifentanil-anesthesia, the patients had a predictably short awakening time, so they could be transferred directly to the general ward. This may, especially in ambulatory surgery, mean cost savings and perhaps higher patient satisfaction. PMID- 12768909 TI - [Cervical teratoma]. AB - We describe the case of a 3-year-old girl with a cervical teratoma. Cervical teratomas are rare tumors in infancy and childhood, commonly presenting as large cervical masses and often diagnosed before birth. In the actual case the teratoma was present at birth but was very small and had a little pseudoduct, which was mistaken for a median cervical cyst. It is emphasized that knowledge of teratomas although often benign in infants and children is essential since they have a malignant potential. If detected early surgical removal is curative provided it is complete. PMID- 12768910 TI - [Picture of the month: skin irritation]. PMID- 12768911 TI - [Estrogen is also necessary for boys and men]. PMID- 12768913 TI - Inflammation and atherosclerosis: mechanisms underlying vulnerable plaque. PMID- 12768912 TI - Ethics and equipoise: rationale for a placebo-controlled study design of platelet glycoprotein IIb/IIIa inhibition in coronary intervention. AB - Rarely is it straightforward to specify the design and parameters of a clinical trial investigating an alternative therapy where effective therapies already exist. If existing therapeutic interventions are highly efficacious, safe, inexpensive, and firmly entrenched, an active-control design becomes the logical first choice. Short of this absolute condition, however, the merits and realities of the scientific, clinical, corporate, and regulatory environments need to be weighed before determining the appropriate approach. A state of clinical equipoise with regard to the use of glycoprotein (GP) IIb/IIIa therapy in percutaneous coronary intervention (PCI) provided the unique opportunity to address the complexities in selecting a placebo-controlled design for the Enhanced Suppression of the Platelet IIb/IIIa Receptor with Integrilin Trial (ESPRIT). ESPRIT investigators assessed whether a high dose of eptifibatide would improve the outcomes of patients undergoing coronary stenting. By using the example of the ESPRIT trial, we examine factors warranting the need for this trial and evaluate the process whereby the United States Food and Drug Administration (FDA) gave approval for a placebo-controlled design. Although the focus of this trial is GP IIb/IIIa inhibition therapy, the issues pertaining to the trial and how they were resolved are general enough to be applied to the design and conduct of clinical trials across a broad spectrum of illnesses and therapeutic modalities. PMID- 12768914 TI - Atherosclerotic plaques: how vulnerable is the definition of "the vulnerable plaque"? PMID- 12768915 TI - Noninvasive visualization of atherosclerotic plaque with electron beam and multislice spiral computed tomography. AB - As an alternative to intracoronary modalities, electron beam computed tomography (EBCT) and multislice spiral computed tomography (MSCT) are able to noninvasively image the coronary arteries. In addition to stenosis detection by imaging the vessel lumen, MSCT has the ability to visualize the coronary artery wall. By using computed tomography (CT), the various components of atherosclerotic plaque may be distinguished and characterized, which holds the promise of, eventually, identifying vulnerable plaque. PMID- 12768916 TI - Vascular MRI in the diagnosis and therapy of the high risk atherosclerotic plaque. AB - Disruption of a high risk plaque is known as the primary cause of cardiovascular events. Characterization of arterial wall components has become an essential adjunct in the identification of patients with plaques prone to rupture. Magnetic Resonance Imaging (MRI) has been revealed as one of the noninvasive tools possibly capable of identifying and characterizing high risk atherosclerotic plaque. MRI may facilitate diagnosis, and guide and serially monitor interventional and pharmacological treatment of atherosclerotic disease. In addition, it permits the simultaneous assessment of the anatomy, morphology, and hemodynamics for the study of flow-induced atherogenesis. It possibly will identify asymptomatic patients with subclinical atherosclerosis. This has potential significance for the improvement of strategies in primary and secondary prevention. PMID- 12768917 TI - Magnetic resonance imaging guided cardiovascular interventions in congenital heart diseases. AB - The purpose of this article is to present some recent applications of diagnostic and interventional MRI in congenital heart disease. To date x-ray-based techniques have been the norm for most diagnostic and therapeutic applications. With the advent of ultrafast MRI and the development of MRI-compatible catheters and guide wires, the goal of achieving real-time guidance by MRI for interventions in congenital heart diseases has proven feasible. We briefly review the latest advances in cardiovascular MRI, and the development of MR-compatible devices for diagnostic and therapeutic applications such as ASD closure and pulmonary artery dilation. PMID- 12768918 TI - A new biological matrix for septal occlusion. AB - The ideal septal occluder scaffold should promote the healthiest and most complete healing response possible while eventually facilitating the full resorption of the material, leaving "native" tissue behind. An excellent biocompatibility of the scaffold tissue is a prerequisite for quick, complete, and firm ingrowth of the device, optimizing outcomes and minimizing the potential for complications. Intestinal collagen layer (ICL) is a highly purified (acellular) bioengineered type-1 collagen derived from porcine submucosa. It is gradually resorbed by the host organism and subsequently replaced by the host tissue. CardioSEAL occluders were modified by substituting the conventional polyester fabric for an intestinal collagen layer (ICL). Percutaneous transcatheter closure of interventionally created atrial septal defects was performed in lambs using these modified occluders. A complete pathomorphological investigation including histology was carried out after 2, 4, and 12 weeks follow up. Standard CardioSEAL implants served as a control group. After 2 weeks in vivo the devices were already covered completely by neo-endothelium. Compared with the conventional synthetic scaffold, ICL devices showed a quicker endothelialization, decreased thrombogenicity, and superior biocompatibility with no significant cellular infiltration observed in the histology of explants with ICL fabrics. After 3 months in vivo the collagen layer remained mechanically intact, but began to show the first histological signs of mild disintegration, gradual resorption, and remodeling. In conclusion, short-term results from preliminary in vivo experiments using a bioengineered collagen matrix as the occluder tissue scaffold showed excellent biocompatibility. This resulted in superior overall results: quicker endothelialization, a decreased thrombogenicity, and decreased immunological host response. PMID- 12768919 TI - Percutaneous closure of mitral paravalvular leak. PMID- 12768920 TI - Advantages and disadvantages of coils for transcatheter closure of patent ductus arteriosus. AB - Different coils have been used to close the patient ductus arteriosus (PDA). In small- and moderate-sized PDA, coils are an adequate alternative to surgery and/or to other devices. The aim of the study is to review and discuss the advantages and disadvantages of using coils (excluding PFM coils PFM Medical, Germany) to close PDA. Cambier was the first to successfully close a PDA using a Gianturco coil. To date, thousands of patients worldwide have undergone transcatheter closure of PDA using this or other types of coils. The use of coils is analyzed with regard to costs in comparison with other therapeutic modalities; techniques--anterograde, retrograde approach, selection of coil size--in relation to the size of the PDA and the available sizes of coils; efficacy of the rate of complete occlusion and the need for reocclusion; and safety in relation to embolization rate, other complications including hemolysis, left pulmonary artery LPA stenosis and coarctation. It is concluded that coils are a cheap alternative for the occlusion of PDA in the small-to-moderate PDA. The technique can be learned quite quickly, it has a high rate of complete occlusion, and has an acceptable rate of safety. The disadvantages include a moderate rate of coil embolization and of hemolysis in patients with residual shunt after coil occlusion in large PDAs. When more than one coil is used, the potential for developing LPA stenosis is high. PMID- 12768921 TI - Extension of transcatheter coil occlusion to the treatment of complex coronary fistulas. AB - Coronary fistulas can be a source of myocardial Ischemia when they are large. In this case series, we describe four complex coronary fistulas treated by transcatheter occlusion. The first two with Gianturco coils and the other two with an Amplatzer PDA device. Transcatheter closure of coronary fistulas can be accomplished in most cases. PMID- 12768922 TI - Interventional strategies in the management of peripheral pulmonary artery stenosis. AB - Peripheral pulmonary artery stenosis challenges therapeutic algorithms for the management of congenital heart malformations. Surgical repair of the proximal pulmonary artery lesion remains with a high recurrence rate while the distal lesions are difficult to access. With the development of transcatheter interventional strategies in the early 1980s, a number of transcatheter treatment options became available. In this review, we summarize the current state of the art for interventional strategies in the management of peripheral pulmonary artery stenosis. PMID- 12768923 TI - Unusual stents in infants with severe congenital heart disease. AB - Two infants with severe congenital heart disease requiring intervention are described. Although both required conventional surgical therapy, mitigating circumstances led to catheter intervention for short-term palliation. The feasibility and advisability of such procedures is discussed. PMID- 12768924 TI - Antitrust implications in nonprofit hospital mergers. PMID- 12768925 TI - Dosage and site of entry influence growth and dissemination of Mycobacterium leprae in T900r mice. AB - The role of dosage of Mycobacterium leprae and the environment of the inoculated site, in producing leprosy lesions in immunologically-suppressed, highly susceptible T900r mice, was investigated. Various doses of M. leprae, i.e., 10(7), 10(6), 10(5), 10(4), were inoculated into both flanks and footpads of two different groups of mice. The sites of inoculation were biopsied for histopathological examination and for M. leprae counts at the end of 6, 8 and 12 months. M. leprae multiplied at the infected site and disseminated [figure: see text] to other parts of the body at all concentrations in the mice that were infected in the footpad with a temperature of 31 degrees C. In animals inoculated at the flanks with a temperature of 37 degrees C, multiplication was recorded only when the dosage of M. leprae was high and there was no dissemination of the organism in any of them. The temperature at the site of entry and the dose of infecting M. leprae may play an important role in the development of leprosy in susceptible individuals exposed to M. leprae. PMID- 12768926 TI - Profiles of the mRNA expression by macrophages infected with Mycobacterium leprae and Mycobacterium avium complex. AB - In the present study, we examined profiles of the interaction of Mycobacterium leprae and Mycobacterium avium complex (MAC) with murine peritoneal macrophages (M phi s) in terms of up-regulation of M phi expression of proinflammatory and immunosuppressing cytolines (CKs) after infection. First, the reverse transcription polymerase chain reaction (RT-PCR) assay revealed that both MAC and M. leprae infections up-regulated M phi mRNA expression IL-12, TNF-alpha, IL-10, and transformating growth factor-beta (TGF-beta), except that M. leprae-infected M phi s showed no increase in the IL-12 mRNA expression. Second, the RT-PCR assay also showed some differences between M. leprae- and MAC-infected M phi s with respect to the modes of IL-10 and inducible nitric oxide synthase (iNOS) mRNA expression. That is MAC, but not M. leprae, infection caused a prolonged increase in the expression of IL-10 and iNOS mRNAs. Third, a ribonuclease protection assay revealed that M phi s co-infected with MAC and M. leprae showed the Il-12, TNF alpha and IL-10 mRNA expression in an intermediate mode of those of M phi s infected with either M. leprae or MAC alone. This implies that the CK expression of M. leprae-infected M phi s may be modified by co-infection with MAC. These findings may suggest differential interactions of M. leprae and MAC organisms with murine peritoneal M phi s in terms of the activation of signal transduction pathways for expression of some kinds of immunoregulatory cytokines and immunoprotective enzymes. PMID- 12768927 TI - Gelatinase activity in Mycobacterium bovis protein extract. AB - Proteases are well-recognized as virulence factors in different pathologies, resulting in tissue damage potential. Despite efforts over the past few years to identify mycobacterial protein antigens, there is little information regarding the role of mycobacterial proteinase activities. In this study, by zymography techniques, we have detected and partially studied some biochemical properties of Mycobacterium bovis proteases, such as pH dependency of activity and susceptibility to classical proteinase inhibitors. We observed optimal proteolytic activity at pH 8. Some proteinases were inhibited by classic inhibitors of serine proteases, such as PMSF, AEBSF, and 3-4 DCI. In some AEBSF pre-treated preparations we observed residual gelatinase activity in Rf 0.32. This gelatinase was stimulated by Zn2+ and inhibited by OPA (1 mM). This last effect was reversed by exposure to equimolar quantitative OPA/Zn+2 (1 mM/1 mM). These results suggest the existence of serine proteinase and metalloproteinase types in protein extracts of Mycobacterium bovis. PMID- 12768928 TI - Saints protectors from leprosy: historical hints of suggestive therapy? AB - A short overview of several saints, venerated in Christian tradition as protectors from leprosy, is offered as an introduction to a more substantial debate on the possibility and sense of analyzing historical sources and the use of their interpretation for modern medical practice. A possible psychoneuroimmunological mechanism has been advanced to relate some of the healings of leprosy reported in historical materials. PMID- 12768929 TI - AP-1 is present in nuclear extracts of lymphocytes from lepromatous leprosy patients. PMID- 12768930 TI - Success of insecticide spraying in controlling malaria. PMID- 12768931 TI - Recurrent meningitis due to unrecognised skull fracture. PMID- 12768932 TI - Need and opportunities for training health professionals in medical genetics. PMID- 12768933 TI - Two mutations in the MTHFR gene associated with mild hyperhomocysteinaemia. PMID- 12768934 TI - SAMA fights 'funder-driven' medical care. PMID- 12768935 TI - PMTCT court order: Manana in contempt? PMID- 12768936 TI - Strikes loom over KZN's salary nightmare. PMID- 12768937 TI - A chance for SA to consolidate on HIV/AIDS. PMID- 12768938 TI - SAMA and TAC march for treatment. PMID- 12768939 TI - Toy gun injuries--more than meets the eye. PMID- 12768940 TI - Homocysteine and the vascular endothelium. PMID- 12768941 TI - Plasma homocysteine and arterial thromboembolic disease. PMID- 12768942 TI - Ethical issues in voluntary HIV testing in a high-prevalence area--the case of Malawi. PMID- 12768943 TI - 'Them and us': scientists and the media--attitudes and experiences. PMID- 12768944 TI - Broken windows of the soul--when is child safety going to grow up? PMID- 12768945 TI - Prescription patterns and utilisation of antihypertensive drugs in private practice in Umtata. PMID- 12768946 TI - Superficial siderosis--case report and review. PMID- 12768947 TI - Intramuscular penicillin is more effective than oral penicillin in secondary prevention of rheumatic fever--a systematic review. AB - BACKGROUND: People with a history of rheumatic fever (RF) are at high risk of recurrent attacks and of developing rheumatic heart disease following a streptococcal throat infection. Giving penicillin to these people can prevent recurrent attacks of RF and subsequent rheumatic heart disease. However, there is no agreement on the most effective method of giving penicillin. OBJECTIVES: To assess the effects of different penicillin regimens and formulations for preventing streptococcal infection and RF recurrence. SEARCH STRATEGY: We searched the Controlled Trials Register (Cochrane Library Issue 2, 2001), Medline (January 1966-July 2000), Embase (January 1985-July 2000), reference lists of articles, and contacted experts in the field. SELECTION CRITERIA: Randomised and quasi-randomised studies comparing: (i) oral with intramuscular penicillin; and (ii) 2- or 3-weekly with 4-weekly intramuscular penicillin in patients with previous RF. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality and extracted data. MAIN RESULTS: Six studies were included (1,707 patients). Data were not pooled because of clinical and methodological heterogeneity of the trials. Four trials (1,098 patients) compared intramuscular with oral penicillin and all showed that intramuscular penicillin was more effective in reducing RF recurrence and streptococcal throat infections than oral penicillin. One trial (360 patients) compared 2-weekly with 4-weekly intramuscular penicillin. Penicillin given every 2 weeks was better at reducing RF recurrence (relative risk (RR) 0.52, 95% confidence interval (CI): 0.33-0.83) and streptococcal throat infections (RR 0.60, 95% CI: 0.42-0.85). One trial (249 patients) showed that 3-weekly intramuscular penicillin injections reduced streptococcal throat infections (RR 0.67, 95% CI: 0.48-0.92) compared with 4 weekly intramuscular penicillin. CONCLUSIONS: Intramuscular penicillin seemed to be more effective than oral penicillin in preventing RF recurrence and streptococcal throat infections. Two-weekly or 3-weekly injections appeared to be more effective than 4-weekly injections. However, the evidence is based on poor quality trials and the use of outdated formulations of oral penicillin. PMID- 12768948 TI - Humalog Mix25 improves 24-hour plasma glucose profiles compared with the human insulin mixture 30/70 in patients with type 2 diabetes mellitus. AB - OBJECTIVE: To compare the effects of Humalog Mix25 (Humalog Mix75/25 in the USA) (Mix25) and human insulin 30/70 (30/70) on the 24-hour inpatient plasma glucose (PG) profile in patients with type 2 diabetes mellitus (T2DM). DESIGN: A randomised, open-label, 8-week crossover study. Study insulins were injected twice daily, 5 minutes before breakfast and dinner. SETTING: Four-week outpatient (dose-adjustment) treatment phase, and 3-day inpatient (test) phase. PATIENTS: Twenty-five insulin-treated patients with T2DM (ages 40-66 years), mean (+/- standard error of the mean) (SEM) HbA1c 7.7% +/- 0.23%, and body mass index (BMI) 29.3 +/- 0.83 kg/m2. OUTCOME MEASURES: 24-hour PG profiles, PG excursions after meals, PG area under the curve (AUC), and 30-day hypoglycaemia rate. RESULTS: The 2-hour PG excursions following breakfast (5.5 +/- 0.34 v. 7.2 +/- 0.34 mmol/l, p = 0.002) and dinner (2.4 +/- 0.27 v. 3.4 +/- 0.27 mmol/l, p = 0.018) were smaller with Mix25 than with 30/70. PG AUC between breakfast and lunch was smaller with Mix25 than with 30/70 (77.6 +/- 3.8 v. 89.5 +/- 4.3 mmol/h/ml, p = 0.001). PG AUC between lunch and dinner, dinner and bedtime, and bedtime and breakfast did not differ between treatments. Pre-meal and nocturnal PG were comparable. The postprandial insulin requirement for lunch meals was supplied equally by the two insulin treatments. The thirty-day hypoglycaemia rate was low (Mix25 0.049 +/- 0.018 v. 30/70 0.100 +/- 0.018 episodes/patient/30 days, p = 0.586) for both treatments. CONCLUSION: In patients with T2DM, Mix25 improved the 24-hour PG profile with lower postprandial PG excursions than with human insulin 30/70. PMID- 12768949 TI - Calcium supplementation to prevent pre-eclampsia--a systematic review. AB - BACKGROUND: Calcium supplementation during pregnancy may prevent high blood pressure and preterm labour. OBJECTIVE: To assess the effects of calcium supplementation during pregnancy on hypertensive disorders of pregnancy and related maternal and child adverse outcomes. DESIGN: A systematic review of randomised trials that compared supplementation with at least 1 g calcium daily during pregnancy with placebo. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register (October 2001) and the Cochrane Controlled Trials Register (Issue 3, 2001) were searched and study authors were contacted. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed. Data were extracted and analysed. MAIN RESULTS: There was a modest reduction in the risk of pre-eclampsia with calcium supplementation (relative risk (RR) 0.68, 95% confidence interval (CI): 0.57-0.81). The effect was greatest for women at high risk of hypertension (RR 0.21, 95% CI: 0.11-0.39) and those with low baseline calcium intake (RR 0.32, 95% CI: 0.21-0.49). There was no overall effect on the risk of preterm delivery, although there was a reduction in risk among women at high risk of hypertension (RR 0.42, 95% CI: 0.23-0.78). There was no evidence of any effect of calcium supplementation on stillbirth or death before discharge from hospital. There were fewer babies with birthweight < 2,500 g (RR 0.83, 95% CI: 0.71-0.98). In one study, childhood systolic blood pressure > 95th percentile was reduced (RR 0.59, 95% CI: 0.39-0.91). CONCLUSIONS: Calcium supplementation appears to be beneficial for women at high risk of gestational hypertension and in communities with low dietary calcium intake. These benefits were confined to several rather small trials, and were not found in the largest trial to date, conducted in a low-risk population. Further research is required. PMID- 12768950 TI - [Keynote lecture: Work seen through the clinical viewpoint: history and ideology]. AB - One of the problems harassing Italy during the period between the 19th and 20th centuries was the "health issue", that was typical of a country penalized by age old backwardness and shaken by the initial shocks of the industrial revolution. Diseases of poverty like pellagra were combined with diseases of progress like those associated with new techniques and relationships in manufacturing. In this regard, Milan developed an ethical and political leadership first in promoting reforms and also in providing the matrix for advanced organization, assistance and scientific models: among them was "work clinicalization" as achieved at the Clinic for Occupational Diseases, founded and directed by Luigi Devoto. PMID- 12768952 TI - [The scientific progress of the "Clinica del Lavoro Luigi Devoto" as seen through the contents of the journal "La Medicina del Lavoro". ]. AB - On occasion of the Centenary of the "Clinica del Lavoro Luigi Devoto", its researchers suggested that awareness of its historical identity and scientific development should constitute the basis for the future of the Institution. The directors of the Institute who have succeeded one to the other over the years have left a vivid report of the vast amount of work developed in several research fields and also of the original results achieved with significant impact on the scientific community and on society. Content analysis of "La Medicina del Lavoro" allowed us to shed light on strong bonds existing between manufacturing techniques, work organization and health, in the different political and social periods in which they developed. Many actions have been defined and several research subjects have been renewed (according to current demands and new methodological possibilities) between the past and the present, nevertheless many new approaches need to be taken in the fields of epidemiology, indoor air quality, ergophthalmology, muskuloskeletal disorders, environmental and molecular toxicology, and further research will be undertaken thanks to the potential of this discipline for the future. The aim of this report is also to express our thanks to all the medical and non-medical staff who have made their contribution over 100 years of engagement. PMID- 12768951 TI - [The centenary of the founding of the "Clinica del Lavoro Luigi Devoto" in Milan (1902-2002). The oldest health organization in the world for the study, treatment, and prevention of occupational diseases]. AB - The foundation of the Clinica del Lavoro by Luigi Devoto cannot be dated back to the specific date of an event or, worse, to the official inauguration as has happened so far. It can rather be considered as a process started on November 20, 1902 (date of the first resolution by the Milan Municipality Council), continuing with the national law n. 365 of July 9, 1905, then with the laying of the first stone on December 11, 1907 bearing the motto of the Milanese School "In aliis vivimus, movemur et sumus" and finally the official inauguration on March 20, 1910. The Founder Luigi Devoto was born in 1864 at Borzonasca (Genoa) and took a degree of medicine and surgery at the Genoa University in 1888. After a period in Prague and then in Genoa, he was appointed temporary teacher of medical pathology in Pavia. He published several scientific contributions in different fields of occupational health (lead intoxication, ankylostomiasis, strain cardiopathy, etc.) and in other branches (tuberculosis, pellagra, etc.) that he considered as indirectly work-related diseases. He died in Milan in 1936. Three issues characterize the ingenious intuition of Luigi Devoto and his thirty-year direction of the Clinica del Lavoro: 1. Full respect for the principles of discipline and scientific method suggested by Bernardino Ramazzini; 2. The extremely broad conception of the diseases that had to be considered as "workers' diseases"; 3. The constant social attention towards the application of scientific knowledge also through workers' information and updating of regulations to improve working conditions. Many of the conflicts and events associated with the foundation of the Clinica del Lavoro can be summarized in the name chosen by the Founder. Actually to those asking why "Clinic of Work" and not "Clinic of Workers", Devoto used to reply that it was work that was ill and therefore it was necessary to treat the work to prevent workers' illnesses. PMID- 12768953 TI - [The Occupational Medicine Post-Graduate School of Milan]. AB - From its very foundation, post-graduate work for medical doctors has been the main objective of the "Clinica del Lavoro". This completion of the doctor's training has, in the course of the past century, moved in time with the evolution of Occupational Medicine as a science: new developments in the field, the results of experimentation and research carried out in the Clinic, have immediately been passed on to young doctors. The professionals trained in the Clinic have always been considered the very best and have been sought after by the most important medical facilities in the country. Since the beginning of the last century, when Luigi Devoto began to evaluate the infectious, poisonous, and deficiency pathologies connected with the workplace, to the post-war years of rapid growth, when Enrico Vigliani studied and prepared our doctors to deal with dust-related pulmonary diseases, metal-based poisons, and many other problems connected with the transformation of an agricultural society to an industrial society, and up to the present years, which have been characterized by the post-industrial new economy, the "Clinica del Lavoro" has always taught and put into practice the motto of Luigi Devoto: "Occupational medicine is not a part of medicine, but rather a particular way to practice medicine". PMID- 12768954 TI - [The role of the "Clinica del Lavoro Luigi Devoto" in international research]. AB - During its long history, the Clinica del Lavoro Luigi Devoto has produced numerous internationally recognized scientific contributions in the specific field of occupational medicine and health, and in other broader areas as, for example, immunology, toxicology, cytogenetics. Pivotal in this activity was the figure of Enrico C. Vigliani, director of the Clinica for over 35 years. We mention here, in particular, the results of the study of "early" markers of lead intoxication; the research on the carcinogenicity of benzene and aromatic amines; the study of the pathogenesis of silicosis and byssinosis; the study of cytogenetic damage after exposure to industrial toxic chemicals; the technical research for dust abatement in mines. Such research also played a major role in the development of pulmonary medicine, toxicology and immunology in Italy, and conferred scientific dignity on the newborn "industrial hygiene" discipline thanks to the work of Nicola Zurlo. Today, the Clinica's research is maintained at international standards especially by the work of thematic Research Centers recently established within the Clinica, covering fields such as occupational and environmental toxicology, occupational oncology, ergonomics, respiratory disease and allergy, psycho-social risk factors. These Centers are also devoted to the development of innovative methods in epidemiology, toxicology, psychology, molecular biology, and ergonomic measurement for the identification and evaluation of health risks at work. PMID- 12768955 TI - [From the past to the future: lessons and expectations. Clinical research in the field of occupational diseases]. AB - The past provides us with important lessons on which to found expectations and future perspectives for our discipline. The scenario of our activities and workplaces is dramatically changing and as a consequence the associated health risks, as well as medical and scientific knowledge on possible effects and impairment and the possibility to document them, the method being however always the same, i.e. the clinical method. Clinical observation leads to the assumption of etiological link and the pathogenetic mechanism of effect, development of adequate diagnostic tools and implementation of prevention, treatment and rehabilitation interventions. Only by sticking to this method, will occupational health be able to continue its function of worker's health protection and continuing education and training of new occupational physicians. In this sense the present and future of clinical research in occupational health is in fact organic development of the lessons of the past which addresses today's work situations using the new potentialities provided by medical knowledge. PMID- 12768956 TI - [Clinical research in the field of occupational diseases (pneumonology aspects)]. AB - In the second half of the seventies, at the same time as the control of silicosis was achieved thanks to pathogenetic and preventive developments, the major efforts of the Clinica del Lavoro in the pneumological field were focused on studying dose-response relationship in asbestos-exposed subjects. Thus for the first time biological indicators (and not radiological ones) were mentioned for asbestos diseases. Bronchoalveolar lavage (BAL) allowed to compare an internal dose indicator (number of bodies/ml) and schlerogenous effect indicators (dose response). Our experience over nearly 20 years for exposure assessment in asbestos-related diseases, based on over 500 cases has produced the following results: parenchymial asbestosis is dose-dependent while pleural plaques are dose independent; epidemiologic data on existence of asbestos resistant subjects were confirmed; asbestos bodies proved to persist in alveolar cavities for several decades. At present, with the ban of asbestos and the increase in forensic medicine cases regarding past asbestos exposures, our Department demonstrated the importance of asbestos body counting in BAL as an objective indicator of asbestos exposure. Besides the asbestos issue, two less frequent work-related diseases were studied and in this regard bronchoalveolar lavage proved to be a substantial diagnostic tool, sometimes even decisive: these were hard metal disease and hypersensitive pneumonitis. In subjects exposed to hard metals, our experience showed the presence of a large lymphocytic alveolitis with CD4+/CD8+ reverse ratio for bronchial asthma and macrophagic-eosinophylic alveolitis characterized by several "bizarre" giant cells in the case of interstitial disease. In extrinsic allergic alveolitis, we noted the presence of a large lymphocytic alveolitis mostly with CD4+/CD8+ reverse ratio, persisting also during clinical remission of the disease. PMID- 12768957 TI - [From industrial hygiene and toxicology to environmental hygiene and toxicology: problems and prospects]. AB - BACKGROUND: Low-dose exposures to mixtures of substances have received increasing interest and they involve many different occupational and environmental situations. The presence in the population (working and general) of groups of susceptible individuals is an important public health issue that poses new challenges to science and society. OBJECTIVES: To discuss the evolution from traditional occupational hygiene and toxicology to the new environmental (general and occupational) hygiene and toxicology. RESULTS: Environmental hygiene and toxicology have remarkably improved analytical tools available to solve most of the analytical issues posed by the present exposure scenario. Biomarkers of low dose exposure, early effects and individual susceptibility are being intensively investigated. CONCLUSIONS: The challenge in this field for the coming years appears to be not the analytical but the medical and ethical implications. PMID- 12768958 TI - [Evolution of industrial toxicology toward vanishing doses and the human genome]. AB - BACKGROUND: This article aims to discuss the influence that the application of the recent discoveries in genomics will have on the theory and practice of industrial toxicology in developed post-industrial countries. It is stressed that the recent advances in toxicogenomics can be integrated into the existing wealth of knowledge on the toxic properties of industrial chemicals to improve the efficacy of prevention of toxicological risk. METHODS AND RESULTS: The understanding of the biochemical and physiological mechanisms underlying susceptibility or resistance to the toxic effects of industrial xenobiotics, and in particular to carcinogens, allows us to split the epidemiologically derived relationship linking the frequency of disease in the exposed population to the level of workplace contamination into a set of sequential sub-relationships linking: a) the exposure level to that of workplace contamination; b) the internal dose to the exposure level; c) the biological effect (e.g., measured through biochemical markers of early effect) to the internal dose; d) the frequency of disease to that of observation of early biochemical effects. Each of the cited relationships is affected by a degree of uncertainty due to the variability of biological response among the examined individuals, which in turn requires a definition of the statistical limits for the association functions between the variables. As a consequence, the possibility of investigating the individual biochemical and physiological steps in the causal mechanism that links toxic exposure to disease does not necessarily lead to an increase in the information potential of biological monitoring, since the uncertainty due to inter-individual variability is amplified through the sequence of causal relationships to the point that the data from biological monitoring become valueless with regard to the prediction of the frequency or probability of disease. This is particularly true when exposure to 'low doses' is investigated, as is now increasingly frequent in post-industrial developed countries, where workplace contamination is now greatly reduced to levels which may be borderline with those in the general environment. Thus at the low-dose end of the range of contamination and exposure values there is an area where, for statistical reasons consequent to the heterogeneity of examined populations, a quantitative prediction of internal exposure due to environmental contamination, of biological adverse effects due to exposure levels and of frequency of disease due to the extent or frequency of biological effects is no longer reliably possible. This in turn impairs the preventive efficacy of biological monitoring. CONCLUSIONS: A closer integration between industrial toxicology and state-of-the-art molecular genetics derived from the recent sequencing of the human genome is the way to overcome the limitations described. In particular, the individual subjects in the examined populations can be classified with regard to some genetically controlled characters relevant to the biotransformation of xenobiotics and to DNA repair and the statistical analysis of data can be performed on more homogeneous subpopulations, in order to decrease inter-individual variability of biochemical and physiological response. This in turn increases the predictive power of the biological markers, both of dose and effect, and improves the efficacy of prevention, e.g., by highlighting oversensitive subpopulations or lifestyles which can increase the risk of occupational and environmental disease. PMID- 12768959 TI - [The experience of the EPM (Ergonomics of Posture and Movement) Research Unit in risk analysis and the prevention of work-related musculo-skeletal diseases (WMSDs)]. AB - The twenty-year experience of the "Ergonomics of Posture and Movement (epm)" Research Unit for prevention of work-related musculoskeletal disorders (WMSDs) is briefly summarized. The epm research unit is the outcome of an agreement between Milan University (Clinica del Lavoro Luigi Devoto), Milan Polytechnic, Don Gnocchi Foundation IRCCS (Bioengineering Centre) and the Regional Health Service (CEMOC of ICP Hospital). Early activities of epm (1985-1993) are first outlined: they are characterized by a wide range of laboratory studies allowing the development of original methods and criteria for on-site analysis of fixed postures and awkward movements and for ad hoc clinical examination of the musculoskeletal apparatus in working populations. Epm contributions are reviewed for the analysis of working activity involving manual load handling (adapted NIOSH method) and hospital patients (MAPO method) as well as for standardization of health surveillance protocols of spinal diseases. Updated contributions are reported on analysis and prevention of upper limb MSDs connected with upper limb repetitive movements (OCRA method). Finally epm's main collaborations with national and international Authorities are summarized as well as the major technical (health promotion) publications addressed to operators and workers, in different working situations, for prevention of musculoskeletal disorders due to biomechanical overload. PMID- 12768960 TI - [Occupational epidemiology: from analysis of the apparent to investigation of the unknown]. AB - This paper, as a contribution for the centenary celebration of the establishment of the "Clinica del Lavoro Luigi Devoto" in Milan (Italy), presents a brief 30 year history of the activities of its Department of Occupational Epidemiology. Studies and methodological contributions that characterized the first decade of activity are presented and grouped under the heading of analysis of known health effects. The second decade was dominated by the studies and activities that originated from the Seveso accident (dioxin), with an initial interest towards molecular epidemiology, which became increasingly relevant during the third decade when we addressed topics like melanoma, lung cancer, and benzene, in addition to dioxin. More traditional occupational approaches were not dismissed and cohort mortality studies are currently under way (textile dyeing and finishing industry, sulfuric acid, tetrafluoroethylene). Pros and cons of the epidemiologic approach are discussed in the context of occupational health and the strength of its methodological apparatus is suggested as a fundamental tool for studying adverse occupational health effects. In contrast, it is stressed how occupational epidemiology has been poorly used in the application of law 626/94. Considering that contemporary epidemiology is much more inclined towards the discovery of new work-related risks (electromagnetic fields, air pollution) than the description of known health effects, the paper suggests that occupational epidemiology enlarge its interests: people and environment outside the factories might be good candidates for study. PMID- 12768962 TI - [Emerging aspects of occupational stress]. AB - Work stress is caused by excessive demands which foster individuals to give prompt cognitive and behavioral answers. When these solicitations exceed the possibilities of the subject to comply, non-physiological reactions may follow, including emotional, neurovegetative and behavioral changes. If adverse stimuli persist, transient alterations form syndromes such as depression, phobic syndromes, anxiety syndromes, hypertension, heart disease, eating disorders, drug addiction, and so on. The authors examine specific stressful working areas causing illnesses in workers such as the burnout syndrome and the mobbing-related adjustment disorder or the post-traumatic stress disorder. PMID- 12768961 TI - [Occupational ophthalmology and ergoophthalmology: a developing field]. AB - In 1979 it was decided to activate, within the Institute of Occupational Health of the Milan "Universita degli Studi", a new Unit of Ergophthalmology. This decision was taken owing to the progressive diffusion of "optical instruments", particularly computer-based equipment, in the world of work and to the relevance that this topic assumed among Italian occupational physicians. Since its beginning, one of the main characteristics of the Ergophthalmology Unit was an interdisciplinary approach, where occupational physicians, ophthalmologists, occupational hygienists and lighting engineers were collaborating on a daily basis. Research activities, which developed over many years and in several phases, were concerned with four main topics: 1. analysis and quantification of "near work load"; initially the investigations were mainly aimed at the evaluation of the observation distance in different tasks, followed by the development of a method which, by means of specifically designed electronic equipment, allows quantification of the accommodation and convergence required by the task; 2. development of a new method aimed at evaluating, by a photometric procedure, the luminance ratios in the "occupational visual field"; this research was carried out, both in the laboratory and during on-site investigations, in collaboration with the Dept. of Electrotechnics of Milan Polytechnic; 3. studies on the relationships between indoor microbial pollution and Computer Assisted Design work, considering that ocular conjunctiva and cornea, due to their external position, can be easily inoculated by micro-organisms present on periocular tissue and hands; this research showed that bacteria and fungi pathogenic for the eye (Staphylococcus aureus, Candida sp., Aspergillus sp.) are present on desk, keyboard and mouse, as well as in the conjunctival sac of the exposed operators; 4. studies on "blue light" and its effects on the Pigmented Retinal Epithelium (PRE) in workers exposed to halide lamps, like electricians, installers, maintenance staff, polymerization operators, etc.; the laboratory and on-site investigation that was carried out in cooperation with the Health Physics Dept. of the Milan ICP Hospital, showed that long-term exposure to low levels of irradiation could constitute an eye risk for these working populations. PMID- 12768963 TI - Malone urges DoH to protect nurses not test them for HIV. PMID- 12768964 TI - Do we need more qualified nurses in outpatients? PMID- 12768965 TI - Does screening save lives? PMID- 12768966 TI - An overview of education for patients with rheumatic diseases. AB - Patient education is central to enabling patients with rheumatic diseases to manage their lives and live as fully and independently as possible. This education can be provided in a range of formats including individual or group sessions, through an arthritis self-management programme and opportunity education, which is informally taught by health professionals. The format selected should be suitable to the individual patient at the particular time it is delivered, and the education should include both topics that health professionals know are relevant and those that patients have cited as being important to them. PMID- 12768967 TI - What you need to know about ... autism. PMID- 12768968 TI - Fluid balance. PMID- 12768969 TI - Care and treatment under the Mental Health Act 1983. AB - The provisions of the Mental Health Act 1983 (for England and Wales) are under review, with a draft bill now published, paving the way for a new act. The bill proposes a much simpler three-step process to replace the current orders for detention and/or treatment, which are discussed here to familiarize nurses with what takes place now. Society has demanded in recent years that changes in society should be reflected in mental health legislation although, to date, there have been many critics of the draft bill's proposals. PMID- 12768970 TI - Implementing a research project on the development of diabetes. AB - The Exeter Research Alliance for Diabetes is a research project recently implemented at the Royal Devon and Exeter Hospital. The project investigates the genetic and environmental influences on the development of diabetes and its associated complications. This article describes the process of instigating the study from an initial research idea through to the implementation of the project. PMID- 12768971 TI - 'This role is close to my heart'. PMID- 12768972 TI - 'A great step forward for nursing'. PMID- 12768973 TI - Pelvic floor muscle exercises for men. PMID- 12768974 TI - Toilet facilities in schools. PMID- 12768975 TI - Consensus document for the use of neuromuscular electrical stimulation. PMID- 12768976 TI - The principles of neuromuscular electrical stimulation. PMID- 12768977 TI - Multidisciplinary continence care. PMID- 12768978 TI - Auditing continence services. PMID- 12768979 TI - Providing integrated services for people with continence problems. PMID- 12768981 TI - [Fellows in Urology in the United States of America]. PMID- 12768980 TI - An evaluation of disposable pads for women with light incontinence. PMID- 12768982 TI - [Henry IV de Castilla( 1454-1474). An exceptional urologic patient. Morphological and personality portrait of Henry IV " The impotent" in contemporary chronicles and manuscripts (I)]. AB - OBJECTIVES: To make an update on Gregorio Maranon's work "Biological essay about Henry IV and his time" (Madrid 1930), reviewing the clinical diagnosis made by the famous spanish doctor-historian more than 70 years ago on the last Monarch of the Castilian Trastamara, because we consider that diseases suffered by Henry IV were urologic mainly. METHODS: We reviewed 10 chronicles, 5 contemporary manuscripts, and 25 works about the figure of Henry IV published up to year 2000. For comparisons between works we evaluated not only the chronicles and works used by Maranon, but also the more recent ones of medical character, which have resulted in a significant increase in the amount of data about the figure of the Monarch. Additionally, following our current knowledge we tried to adapt the clinical diagnosis from Maranon--eunucoid dysplasia with acromegalic reaction--to current nomenclature in order to define the kind of endocrinopathy the Monarch suffered. RESULTS: Following a review of contemporary chronicles and recent bibliography about Henry IV, the Monarch was a singular urologic patient. He suffered chronic renal lithiasis, impotence, a presumptive penile anomaly, and almost surely infertility, although all his uro-andrological diseases were part of the endocrinopathy the Monarch suffered, probably a hypophyseal tumor. Included in the work there are descriptions of his physiognomy and psychopathological characters of his personality from the chronicles, commenting on all clinical symptoms the Monarch suffered according to his biographers and present day doctors and historians. PMID- 12768983 TI - [Henry IV of Castilla (1454-1474). An exceptional urologic patient. An endocrinopathy causing the uro-andrological problems of the Monarch. Chronic renal lithiasis (II)]. AB - OBJECTIVES: To review all known data about the endocrinopathy and renal disease suffered by Henry IV of Castile according to contemporary chronicles and manuscripts, comparing the clinical diagnosis made by Gregorio Maranon 70 years ago in his work "Biological essay about Henry IV and his time" (Madrid 1930) with present concepts, because we consider, as other medical authors, that his endocrinopathy is not well determined. Regarding his chronic renal lithiasis, it could have played an important role as negative factor in the Monarch's quality of life, and have been related to his endocrinopathy, so that it should also be reviewed. METHODS: We reviewed a total of 10 chronicles and 5 contemporary manuscripts treating the look and diseases of Henry IV to obtain the most important data about both diseases and be able to characterize the kind of endocrine disease suffered by the Monarch, and all aspects regarding his lithiasis, and compare them with our current knowledge. RESULTS: From the review of those chronicles and manuscripts, it is my opinion that Henry IV suffered acromegaly secondary to a GH and prolactine producing hypophyseal tumor from childhood, which might justify the impotence he suffered from his youth and other symptoms clearly referred in the chronicles. Chronic renal lithiasis (flank pain, lumbar illness, and hematuria) finally led to acute obstructive uropathy, main cause of his death. This fact has not been emphasized by historians. The renal lithiasis benign part of a multiple endocrine neoplasia syndrome (MEN I) can not be ruled out. PMID- 12768984 TI - [Henry IV of Castilla (1454-1474). An exceptional urologic patient. An endocrinopathy causing the uro-andrological problems of the monarch. Impotence and penile malformation (III)]. AB - OBJECTIVES: To review, in accordance to contemporary and current chronicles and manuscripts, all data known about impotence and presumptive penile malformation suffered by Henry IV of Castile. Although currently the monarch's homosexuality is almost totally rejected, because according to many authors slandering, mainly of the sexual kind, was used as a political arm to discredit him by his enemies, it is yet to be demonstrated if his erectile dysfunction and presumptive genital anomaly were also slanderly disseminated by his detractors to avoid recognizing his daughter Juana "La Beltraneja" as successor to the throne. METHODS: We reviewed a total of 10 chronicles, 5 contemporary texts and manuscripts, and 25 books about the figure of Henry IV published before year 2000, analyzing medical works in detail, particularly those from Maranon, Eisenberg, and Irvine and Mackay, who studied both pathologies trying to differentiate if both diseases could be justified by the kind of endocrinopathy the monarch suffered. RESULTS: Although it is understandable that the study could be mistaken by document manipulation undertaken during Henry IV's life and posteriorly during the reign of the "Catholic Kings", and due to the lack of impartiality of some of his chroniclers, it is my opinion that the most probable cause of Enrique IV's erectile dysfunction was a GH and prolactine producing hypophyseal tumor, that he suffered from his youth producing complete impotence and acromegaly. It is not well documented if the Monarch also suffered a congenital penile anomaly. PMID- 12768985 TI - [Henry IV of Castilla (1454-1474). An exceptional urologic patient. An endocrinopathy causing the uro-andrological problems of the Monarch. Artificial insemination attempts (IV)]. AB - OBJECTIVES: To analyze, in accordance to contemporary and current chronicles and manuscripts, all known data about possible infertility or sterility suffered by Henry IV and attempts of artificial insemination undertook by his wife Juana of Portugal with the Monarch's semen due to his complete impotence. His possible infertility was unfairly used by his enemies to deny the right of succession to his daughter Juana, called "La Beltraneja", in a similar way than what has happened with his erectile dysfunction. In our work, we tried to demonstrate if Henry IV was or not able to conceive and paternity of his daughter Juana, based on our current knowledge and presumed endocrinopathies suffered by the Monarch in accordance to some medical writers. METHODS: We reviewed a total of 10 chronicles, 5 contemporary texts and manuscripts, and 25 books about the figure of Henry IV published before year 2000, analyzing medical works in detail, particularly those from Maranon, Eisenberg, and Suarez, who studied the facts referred to the presumed Monarch's infertility and his paternity trying to establish a relationship with current endocrinologic syndromes that cause infertility or sterility and with the chronological dates of conception and birth of Juana "La Beltraneja". RESULTS: As it has been said before, although the study may be falsified by document manipulation done by those chroniclers partisans of the Catholic Kings, retrospective analysis seems to show that if the Monarch suffered eunuchism (hypogonadism) or hypophyseal tumor he should have been sterile. In a similar way than Suarez, we don't dare to judge if the attempts of artificial insemination performed to his wife Juana were successful or not, so the historical controversy about "La Beltraneja" continues. PMID- 12768986 TI - [Testicular Lesions not requiring treatment]. AB - OBJECTIVES: To review a group of testicular lesions either clearly benign or that may simulate malignant neoplasia on testicular ultrasound. To describe those clinical and ultrasound findings allowing to make the right differential diagnosis, and proper clinical management. METHODS: We collect a numerous series of cases seen in the Urological Radiology Unit at the University Hospital "Marques de Valdecilla", Santander (Spain), and evaluated by means of a Logiq 500 GE ultrasound. For this study we used linear multifrequency probes (6-11 mHz). RESULTS: A series of patients with benign testicular lesions are presented. The most characteristic ultrasound findings in the grey scale, color-Doppler and Angio-Doppler are reviewed. CONCLUSION: Ultrasound is the diagnostic tests of choice in the study of scrotal pathology. This technique allows easy differentiation of lesions within the testicles from extra testicular ones. Such a simple difference is of vital value because it is demonstrated that most intra testicular masses are malignant, whereas extra testicular lesions are generally benign. However, experience has demonstrated us that orchyectomies resulting in a benign histological process were the few, although significant. We describe various benign testicular lesions with characteristic ultrasound features. These features in association with patient's clinical picture and negative tumor markers allow surgery avoidance in a given patient. Then, it is a question of knowing and identifying these lesions, and differentiating them from malignant neoplasia lesions with the purpose of avoiding unnecessary surgical interventions. PMID- 12768987 TI - [Comparative analysis of complications between different surgical techniques in the treatment of urinary stress incontinence at our centre]. AB - OBJECTIVES: Surgical repair is the most effective treatment for stress urinary incontinence (SUI) currently. Nevertheless, this method is not without complications. The objective of this work is to compare the incidence of complications between different groups of surgical techniques employed for the treatment of SUI in our patients. METHODS: We performed a retrospective analysis of patients diagnosed of SUI who underwent surgery at our department between January 1991 and December 1999, grouping patients in three categories depending on the procedure: abdominal technique, abdomino-vaginal techniques, and sling procedures. Complications were grouped into two categories: major and minor complications. Chi-square and Fisher's test were used for the association analysis of complications' frequencies. All confidence intervals and level of statistical significance were calculated for a p value < 0.05. RESULTS: 5.16% of the patients undergoing surgery for SUI suffered major complications. Globally, the most frequent complications were suprapubic pain (33%), wound seroma and/or infection (20%), postoperative acute urinary retention (AUR) (26%), and presence of significant post void residual (24%), being the sling procedure with a greater number of postoperative complications, as much in postoperative pain, as in AUR, as in presence of post void residual (p < 0.05). CONCLUSIONS: Sling procedures present a significantly higher percentage of complications when compared with abdominal and combined techniques. PMID- 12768988 TI - [Comparative study of P 53, Bcl-2, and C-erbB-2 expression in low-grade papillary bladder tumors]. AB - OBJECTIVE: To study the prognostic value of p53, bcl-2 and c-erbB-2 immunoexpression in predicting tumor relapses in low-grade papillary bladder neoplasms. METHODS: In all patients a complete transurethral resection of the lesion was performed. All the material was formalin-fixed and paraffin-embedded. At the immunohistochemical level, the following monoclonal antibodies were utilized: p53 (clone DO7), bcl-2 (clone 124) and c-erbB-2 (clone CB11). In order to predict tumor relapses during follow-up, a study of specificity, sensitivity and predictive positive value (PVP) and negative was designed. In univariate statistical studies, the following tests were utilized: Chi-square, Kaplan-Meier estimates and Cox logistic regression. RESULTS: Mean follow-up was 76.6 months (38 to 168). In recurrence prediction, p53 expression showed a high specificity (99%) as well as a high PPV (96%). Regarding bcl-2 and c-erbB-2 immunoexpression, both specificity (65% and 72%) and PPV (61% and 72%) were also high, although these percentages were lower than those obtained for p53 expression. The combined analysis of p53 and bcl-2 indicated that bcl-2 immunoexpression in non-basal cells of the urothelium could be independent of p53, although the number of cases showing this particular expression pattern is not high enough to perform an accurate statistical analysis. Otherwise, histologic grade demonstrated higher sensitivity (64%) and lower specificity (40%) than the immunohistochemical markers. In univariate studies, p53 showed an intense statistical correlation with relapse-free interval (RFI) and prediction of tumor recurrences during follow-up (p < 0.001), whereas the expression of bcl-2 (p = 0.065) was nearly correlated with RFI (p = 0.065). In contrast, expression of c-erbB-2 did not show statistical correlation (p = 0.112). CONCLUSIONS: In our study, individual and combined analysis of p53 and bcl-2 immunoexpression have demonstrated to be useful in predicting tumor recurrences and RFI in low-grade bladder lesions. PMID- 12768989 TI - [Laparoscopic radical prostatectomy. Review of our first year]. AB - OBJECTIVES: To present our series of laparoscopic radical prostatectomy, comparing to the published results up to date. METHODS: From november 2001 to november 2002, we have practiced 27 laparoscopic radical prostatectomies without lymphadenectomy. We have used the Montsouris technique with some modifications. All cases started by laparoscopy are included in the report. RESULTS: Laparoscopic approach was indicated in 27 patients; 17 cases were completed and 10 converted to open surgery. Among the last 17 consecutive cases only two were converted. Mean operative time was 296 minutes (190-480 minutes, MAY: SD 60). Global morbidity was 18.5%, without severe complications, reoperations or mortality. Transfusion rate was 11%. There was not any incontinent patient and three patients developed an anastomostic stenosis. 57% of previously potent patients have erections with a follow-up over six months. Positive surgical margins incidence is 33% overall and 17.6% in laparoscopic cases. Only one of the patients with a follow-up above six months has a biological (PSA) recurrence (7%). CONCLUSIONS: Laparoscopic radical prostatectomy is feasible, reproducible and within reach of interested urologists. It may substitute, once sufficient experience is acquired, conventional surgery. PMID- 12768990 TI - [Renal colic in a patient with crossed renal ectopia]. AB - OBJECTIVES: To report a new case of renal colic in a patient with crossed renal ectopia and L-shaped kidney, and to perform a bibliographic review. METHODS AND RESULTS: A 33-year-old male patient consulted for abdominal right pain. Ultrasound and CT showed a L-Shaped Kidney with left renal and ureteral stones. These stones produced obstructive left collecting system. This patient was managed with left pyelolithotomy and ureterolithotomy. The transabdominal approach was recommended to resolve this stones. CONCLUSIONS: The crossed renal ectopia with fusion is a rare condition. The abnormal kidney position and the anomalous kidney supply may impede drainage from the collecting system, creating a predisposition to urinary tract infection and calculus formation. The colic in this patient sometimes is misdiagnosed how abdominal disease. The usual method of detection was excretory urography but now ultrasonography and CT have showed more cases. The indications for open surgery in treatment the stones, are generally failure of extracorporeal shock wave lithotripsy, this is common in these cases for the anatomic abnormality, and the transabdominal approach is recommended to resolve its complications in this kidneys. PMID- 12768991 TI - [Papillary cystadenoma of the epididymis: Report of 2 new cases]. AB - OBJECTIVE: To report two cases of papillary cystadenoma of the epididymis. METHODS: Clinical and pathological study of 2 male patients 48 and 26-year-old respectively, presenting with a solid, painful mass in the head of the epididymis. The nodules were removed. RESULTS: Pathological analysis showed two papillary cystadenomas of the epididymis. CONCLUSIONS: Papillary cystadenoma of the epididymis is an epithelial tumor of unclear origin. It is associated with von Hippel-Lindau disease and infertility. They are uncommon and benign. PMID- 12768992 TI - [Urachal cysts during childhood]. AB - OBJECTIVE: To assess diagnosis and treatment of uracal cysts. METHODS: We reviewed the medical records and radiological studies of four patients who were treated for a urachal anomaly: two urachal cysts and two urachal abscesses. Diagnostic evaluation included, ultrasound, computerized tomography and magnetic resonance. RESULTS: Treatment of urachal abscesses involved drainage and secondary excision. The urachal cyst does not require surgical intervention. CONCLUSIONS: Ultrasound is an excellent diagnostic tool for patients with urachal cysts. Infected cysts may present with pain, tenderness and erythema in the infraumbilical region. Extraperitoneal excision is the treatment of choice, although large infected cysts may require an initial drainage procedure. PMID- 12768993 TI - [Juvenile gangrenous vasculitis of the scrotum]. AB - OBJECTIVE: To describe clinical and pathological findings of a new case of juvenile gangrenous vasculitis of the scrotum, a peculiar and unusual (only 7 cases, approximately, have been previously reported) type of scrotal gangrene described by Pinol in 1973. METHODS: A 24 year-old male presented at the emergency room with a right scrotal necrotising cutaneous lesion and fever. Physical examination, complete blood count, biochemical test, roentgenographic abdominal and thoracic findings, microbiological test and biopsy were performed. RESULTS: Clinical aspects (symptoms and sings) and results of complete blood count (leukocytosis), normal ultrasound exploration and biochemical and roentgenographic abdominothoracic findings, microbiological test (negative) and biopsy (necrotising vasculitis) are compatible with juvenile gangrenous vasculitis of the scrotum. CONCLUSIONS: Juvenile gangrenous vasculitis of the scrotum is a rare entity which should be diagnosed by strict clinicopathological diagnostic criteria because treatment and prognosis are distinct of other numerous ulcerative scrotal diseases. PMID- 12768994 TI - [Spontaneous pre-vesical haematoma in a patient under anticoagulant treatment]. AB - OBJECTIVES: To report a case of perivesical hematoma without previous trauma in a patient under anticoagulant treatment. METHODS: A 70-year-old female admitted with a transitory ischemic cerebrovascular accident was given sodium heparin. Four days later she presented with abdominal distention and hypogastric discomfort. Abdominal ultrasound showed a big perivesical hematoma. RESULTS: CT scan confirmed the presence of a big hematoma. The hematoma was drained five days later after an optimal coagulation profile. Patient was discharged with normal ultrasound after an uneventful postoperative course. CONCLUSIONS: Spontaneous bleeding in patients with coagulation abnormalities is a well-known event, although there are few bibliographic references about primary perivesical blood collections. We opted for deferred drainage of the hematoma to avoid perivesical fibrosis. PMID- 12768995 TI - [Bladder hemangioma after radiotherapy]. AB - OBJECTIVES: Bladder hemangiomas are exceptional benign tumours representing 0.6% bladder tumours. Most times they present as solitary, unique lesions; radiological signs are very similar to urothelial neoplasias making differential diagnosis or at least diagnostic suspicions very difficult due to its low incidence. We perform a bibliographic review and comment on its clinical and pathological characteristics, as well as on diagnostic methods and latest modifications in therapy. METHODS: We present a case of bladder haemangioma secondary to pelvic radiotherapy, being this presentation unique among approximately 100 cases described in the literature. RESULTS: From cystoscopy we suspected the lesion could be an endothelial neoplasia based on its vascular appearance. We performed transurethral resection and electrocoagulation of the lesion because it was small and unique. Pathology report confirmed it was a bladder hemangioma secondary to radiotherapy, being this an exceptional presentation. There has been no relapse up to date. CONCLUSION: We think that resection and electrocoagulation of unique, solitary bladder hemangiomas is a therapeutic option to be taken into consideration in the management of this kind of benign tumours. PMID- 12768996 TI - [Extra adrenal pheochromocytoma. Report of two cases]. AB - OBJECTIVES: To review two cases of extra adrenal pheochromocytoma. METHODS: We present two cases: a 75-year-old patient with history of high blood pressure and intravesical tumor compatible with urinary bladder paraganglioma, and a 59-year old patient with a retroperitoneal tumor arising from para-aortic ganglia. RESULTS: Diagnosis was made by CT scan and transurethral resection in the first case, all posterior extension studies were negative; in the second case retroperitoneal exeresis surgery was performed after CT scan. CONCLUSIONS: Extra adrenal pheochromocytomas are catecholamine producing tumours, which determines their clinical features; treatment should be surgical. A bibliographic review about this disease is performed. PMID- 12768997 TI - Recent data upon impotence, incontinence and quality of life issues concerning radical prostatectomy. AB - OBJECTIVE: To present recent data dealing with the two major post-radical prostatectomy complications incontinence and sexual dysfunction, and investigate if they affect the quality of life of the patients. METHODS: We evaluated recent data available on Medline, from different series examining the incidence, pathophysiology, evaluation and treatment of incontinence and impotence after radical prostatectomy. We also evaluated the effect upon quality of life of these complications. RESULTS: Many experienced surgeons observe low incidence rates of incontinence and sexual dysfunction after radical prostatectomy and it seems that meticulous surgical technique might prevent these devastating complications. However, there are many papers in the literature, which show a great variety of results with incontinence ranging from to 0.3 to 65.6% and potency rates from 11.6 to 86% among treated patients. Depending on the individual's opinion incontinence is a minor problem and does not have a great impact upon their quality of life. Impotence affects more frequently these men and seems to be the most disturbing complication after radical prostatectomy affecting the quality of life. CONCLUSIONS: Today avoidance of major complications after radical prostatectomy depends mostly upon high quality surgical technique. Post-radical prostatectomy incontinence does not seem to be a frequent event and when it occurs it does not dramatically affect patients' quality of life. Impotence although most annoying for the majority of the treated patients can be effectively managed by various methods when the neurovascular bundles are preserved. PMID- 12768998 TI - Stability of risk factors for cardiovascular diseases in Portuguese children and adolescents from the Porto area. AB - An important aspect of preventive medicine is to identify subjects at risk as soon as possible, so preventive strategies can be introduced at early ages. The justification for this strategy is twofold: firstly, the assumption that children maintain a particular high value of a risk factor for disease throughout life; and secondly, the assumption that lowering the level of the risk factor in early life will have a greater impact on the disease than will risk factor changes in later life. In epidemiology the analysis of such factors over time is referred to as tracking. Tracking analysis has been applied to risk factors for cardiovascular diseases (CVD) in pediatric years. The aims of this study were: I) to analyze the stability of biological risk factors [high blood pressure (BP), high percentage of fat mass (%FM) and high total cholesterol (TC)] and lifestyle risk factors [low physical activity index (PAI)] in isolation; and II) to analyze the stability of zero, one, two or three biological risk factors. There were two evaluations in 692 children and adolescents (325 boys and 367 girls), aged between 8 and 15 years. The quartiles, adjusted for age and gender, were the criterion used to identify subjects with biological risk factors (fourth quartile) and with lifestyle risk factors (first quartile) for CVD. The stability was calculated through the relative frequency of subjects who maintained or changed quartile between the two evaluations. There is stability for biological risk factors as well as for behavioral and/or lifestyle risk factors. However, the highest stability is seen in biological risk factors. PMID- 12768999 TI - Serum ferritin levels and other indicators of organic iron as risk factors or markers in coronary artery disease. AB - BACKGROUND: Several international studies have discussed whether serum ferritin is a risk marker in coronary heart disease. The objective of this study was to evaluate the importance of serum ferritin levels and other indicators of organic iron as possible risk factors or markers in coronary artery disease. Secondly, the classical factors were studied in order to identify possible associations with organic iron markers. METHODS AND RESULTS: In a medical institution, 1263 patients underwent cinecoronary arteriography from December 1997 to May 1998. A sample of 400 patients was separated, at random, to establish a comparative clinical study between two groups: group A, comprising 200 individuals with coronary atherosclerosis and group B, comprising 200 patients without coronary atherosclerosis, as confirmed by cinecoronary arteriography. From group A, 182 patients (130 males) and from group B, 157 (96 females) did not show any exclusion criteria and were considered eligible. All women were in the postmenopausal period. The blood samples were collected by a biologist, between 8.30 and 9.0 a.m., after a 12-hour fast and a 36-hour non-smoking period. In order to analyze all results, univariate analysis, the logistic regression technique and the interactive forward stepwise method were used in order to optimize the model and to predict the chances of coronary atherosclerosis. The results of the logistic regression with all the variables analyzed showed that male gender, age, smoking, triglycerides/VLDL interaction, increased serum LDL-C levels and decreased serum HDL-C levels are important to predict the chances of coronary atherogenesis. CONCLUSION: Serum levels of ferritin and of other organic iron indicators--transferrin saturation, total iron-binding capacity, hemoglobin and hematocrit--were neither risk factors nor risk markers for coronary atherosclerosis. Paradoxically, serum iron levels were higher in the group without atherosclerosis. In this study, variables classically considered as risk factors were similar to those in the literature. PMID- 12769000 TI - Prognostic impact of peak oxygen uptake and percentage of predicted value of peak oxygen uptake in patients with left ventricular dysfunction. AB - BACKGROUND: Comparing the prognostic value of peak oxygen uptake (VO2max) in absolute values and percentage of predicted VO2max (%PredVO2) in terms of prognostic stratification, contradictory references are found in the medical literature. In theory, the measurement of VO2max relative to a normal subject should improve its predictive capacity, because VO2max is influenced by gender, weight, physical activity and age. OBJECTIVES: To compare the prognostic value of VO2max and %PredVO2 (Jones equation) in an adult population (> or = 20 years) of patients with left ventricular systolic dysfunction and to study the influence of age on these parameters. POPULATION AND METHODS: We review data from the first cardiopulmonary stress test performed in 295 consecutive patients with left ventricular systolic dysfunction (ejection fraction < or = 40%); mean age was 52 +/- 10 years, 81% were male, and 76 aged > or = 60 years. In 52%, the etiology was ischemic. We evaluated demographic and clinical variables, the baseline ECG and the medication used before the test. We compared VO2max (ml/kg/min) and %PredVO2 in terms of prognostic stratification for the combined endpoint (death and/or heart transplant) for 2 years of follow-up. RESULTS: We detected 60 patients with events (42 deaths and 18 transplants). ROC curve analysis was used to determine the best cut-off value for VO2max and %PredVO2 to identify patients with events. The best cut-off for VO2max was < or = 19 ml/kg/min and < or = 49% for %PredVO2 in the total population, and in the event-free survival analysis, by log-rank test, we obtained p < 0.001 for both cut-offs. Considering the two extreme age groups (20-40 years and > or = 60 years), we found different cut-off values. In the youngest group we obtained VO2max of < or = 23 ml/kg/min and %PredVO2 of < or = 59%, and in the elderly < or = 12.5 ml/kg/min and < or = 64% respectively. For the intermediate group, the cut-off values were the same as for the total population. In the youngest patients, the new cut-offs did not offer an improvement in accuracy. In the elderly, for VO2max, we obtained an improvement in specificity from 28 to 86% and a reduction in sensitivity from 95 to 50%, with an improvement in accuracy from 46.8 to 75.9% (p = 0.05). With regard to %PredVO2, specificity fell from 86 to 61% and sensitivity increased from 50 to 82%, with similar accuracy (75.9 to 67%, p = 0.78). CONCLUSIONS: In the extreme age groups, we obtained different cut-off values, but only for the elderly (VO2max < or = 12.5 ml/kg/min and %PredVO2 < or = 64%). A benefit in terms of accuracy was only obtained in respect of VO2max, because for %PredVO2, there is already a correction for age through the Jones equation. PMID- 12769001 TI - Intracardiac thrombosis and mitral prosthesis dysfunction in systemic lupus erythematosus. A case report. AB - Systemic lupus erythematosus (SLE) is an inflammatory disease characterized by deposition of autoantibodies and pathogenic immune complexes in the cells and tissues of an organism, causing lesions. The heart is one of the most frequently involved organs, valvar involvement being common (27%) and of great clinical importance. Some studies suggest that valvar involvement is associated with the presence of a specific autoantibody, the antiphospholipid antibody (aPL). In these circumstances, major complications may occur such as arterial or venous thrombosis, thrombocytopenia, repeated spontaneous abortions and presence of lupus anticoagulant antibody. Among the semiologic methods for cardiac evaluation, echocardiography is the main instrument for investigating the anatomical and functional involvement of the heart in the great majority of systemic diseases like SLE. We report the case of a patient with SLE and aPL syndrome who underwent surgery for mitral valve replacement with a biological prosthesis and died nine months later due to massive intracardiac thrombosis and prosthesis dysfunction. PMID- 12769002 TI - Hyperhomocysteinemia--case report. AB - The authors present the case of a 25-year-old female patient, white, with mental retardation and proptosis, and a history of repeated cerebrovascular events. During investigation elevated levels of homocysteinemia and homocystinuria were demonstrated. The authors present a review of related literature. PMID- 12769003 TI - Mitochondria: role in ischemia, reperfusion and cell death. AB - Recent advances in the knowledge of the biochemical basis of myocardial ischemia have enabled a better understanding of the complex sequence of events occurring in ischemic cardiomyopathy, whatever its manifestations. This has clearly highlighted the important role played by cardiac mitochondria in these events. At first only associated with energy production, mitochondria have been clearly shown to have other important functions, like the maintenance of calcium homeostasis, as well as ischemic and non-ischemic preconditioning, and also modulation of cellular life and death. The aims of this review are twofold: firstly, to review the current knowledge on mitochondrial morphology and structure, and how these can be affected by ischemia and ischemia-reperfusion; and secondly, to summarize the role of cardiac mitochondria in cardioprotection and modulation of cell death mechanisms. PMID- 12769004 TI - Applicability of clinical trial results to the individual patient: practice guidelines. AB - Randomized controlled trials (RCTs) constitute the methodological gold standard to generate therapeutic or preventive evidence. Diagnostic or prognostic evidence is given by studies with different designs, rather than RCTs. Given the structural design that a RCT must present to be valid--discussed in previous issues of the Journal--one major question is the applicability of trial results to populations or individual patients that are different from those analyzed in that trial. In other words, how does the cardiologist know whether to apply a specific treatment--whose efficacy and risk profile have been defined in a good quality RCT, i.e. one with valid and important results--to his/her individual patient? The simplest solution would be found in the answer to the question: "Could my patient have been enrolled in the study by satisfying its inclusion and exclusion criteria?" If the answer is "yes", then it would be sound to apply the study results to the patient, so long as it is done carefully. If the answer is "no" (because the patient presents different demographic features or serious comorbidities not found in the trial subjects, for example), then transfer of the therapeutic results from the trial would be seriously compromised. The evaluation of these questions is complex and, due to its importance, deserves a systematic approach. In this article we will present some practical guidelines on whether to reject the applicability of RCT results to patients who are different from those included in the trial. PMID- 12769005 TI - Portuguese Association of Arrhythmology, Pacing and Electrophysiology (APAPE) 2001 national registers of interventional electrophysiology. PMID- 12769006 TI - An unusual case of aortic stenosis. PMID- 12769007 TI - Echocardiographic demonstration of total percutaneous closure of patent ductus arteriosus. PMID- 12769008 TI - Legg-Calve-Perthes disease: one century later. PMID- 12769009 TI - Incorporation of massive bone allografts: can we achieve better performance? PMID- 12769010 TI - Solitary enchondromas: is radiographic follow-up sufficient in patients with asymptomatic lesions? AB - Enchondromas are benign cartilaginous tumours and rarely transform into chondrosarcomas. Curettage is usually performed, and a low rate of complications is assumed. We analysed retrospectively data from 73 patients with enchondromas treated by curettage with respect to symptoms, therapy, complications and recurrences. All patients were treated by curettage of the tumour, in most cases followed by cancellous bone grafting. Twenty-three percent of the patients presented a complication. There were two recurrences but no malignant transformations in the follow-up period. During the same time period 29 patients with chondrosarcomas were treated, including two secondary chondrosarcomas. One had a recurrence of a benign enchondroma of the metacarpal and developed a pulmonary metastasis from a chondrosarcoma. The other was a patient with Ollier's disease who had a secondary chondrosarcoma of the radius. Malignant transformation of a solitary enchondroma to a chondrosarcoma is rare. On the other hand the complication rate of enchondroma curettage is considerable. Regular radiological follow-up of asymptomatic enchondromas may therefore be the better option. PMID- 12769011 TI - Superior efficacy in suppression of heterotopic bone formation using fractionated irradiation of 5 x 2 Gy compared to a single dose of 7 Gy. An experimental study in rats. AB - Postoperative irradiation of the operative field is an established method to prevent heterotopic ossification in total hip arthroplasty. In this study two theoretical dose-equivalent regimens of radiation therapy were compared. Allogenic bone matrix was implanted in both thighs of 50 adult male Wistar rats to induce heterotopic ossification. Immediately after operation the implants of 40 animals were irradiated using a single-dose of 7 Gy or 5 fractions of 2 Gy each. Ten rats served as a controlgroup and did not undergo irradiation. Radiation therapy with 5 x 2 Gy led to a highly significantly better suppression of heterotopic ossification than irradiation with 1 x 7 Gy (p < 0.001; paired-t test). Single-dose irradiation reduced the mean calcium contents to 138.87 +/- 22.84 micrograms Ca2+/mg implanted bone matrix; fractionated irradiation obtained a reduction to 63.35 +/- 21.16 micrograms Ca2+/mg implanted bone matrix. In thigh implants not exposed to irradiation the mean calcium content was 191.50 +/- 11.46 micrograms Ca2+/mg implanted bone matrix. Radiographically better suppression of bone formation could be documented after irradiation with 5 x 2 Gy compared to 1 x 7 Gy and non-irradiated implants. The histological aspect of the explanted specimens showed quantitatively more new bone formation in the non-irradiated controls than in both irradiation groups. In view of experimentally demonstrated better effects, as well as the reduced side effects, fractionated irradiation appears preferable. PMID- 12769013 TI - Shoulder and elbow function following Marchetti-Vicenzi humeral nail fixation. AB - Humeral nailing has been associated with reduction of shoulder and elbow function. We present the results of fixation of 15 diaphyseal humeral fractures with Marchetti-Vicenzi nails (B Braun Medical Aesculap). Shoulder and elbow functions were assessed by the Constant shoulder and Mayo elbow scoring systems. The average Constant Shoulder score was good (82.5/100) and the average Mayo Elbow Score was excellent (95.4/100). The indications for surgery included nonunion, pathological fractures and poor fracture position. The mean follow-up was 67 weeks with an average interval to surgery of 19 weeks. The mean time to union was 19 weeks. All the fractures united without any patient requiring any further procedure. There were two postoperative radial nerve palsies and one median nerve palsy, however two patients had complete recovery and one had partial recovery of the radial nerve. There was one deep infection requiring removal of the implant. There were no implant failures. Based on our experience, the Marchetti-Vicenzi humeral nail appears as a relatively safe implant and its use has been associated with preservation of good shoulder and elbow functions. PMID- 12769012 TI - Discriminating factors for functional outcome after shoulder arthroplasty. A critical review of the literature. PMID- 12769014 TI - Repeated grip strength at one month interval and detection of voluntary submaximal effort. AB - This study was set up to test our hypothesis that using the Jamar dynamometer a voluntary submaximal effort has a greater variability compared to a real maximal effort, especially when there is a long interval between different measurements. We tested 32 volunteers without a history of upper limb injury or operation in our hospital, with an interval of four to five weeks between tests. Each volunteer was asked to grip first right-handed then left-handed, first with a real maximal effort and then with a submaximal effort. This test was performed twice during the same session. The same individuals were seen again one month later and were studied using the same protocol. Sensitivities and specificities for the grip test were calculated using six different criteria to indicate a submaximal effort. Repeated grip testing with a time interval of one month cannot reliably detect voluntary submaximal effort in healthy volunteers. PMID- 12769015 TI - Hip fracture treatment: outcome and socio-economic aspects. A one-year survey in a Belgian University Hospital. AB - Between October 1, 1997 and September 30, 1998, 201 consecutive hip fractures in patients over the age of 50 were registered according to the SAHFE (Standardised Audit of Hip Fractures in Europe) protocol. The mean age was 81.3 years; 75% were females, more than 40% were admitted from an institution and fewer than 10% were completely fit. Almost 60% of the fractures occurred in the trochanteric region while less than 40% were intracapsular. All fractures but one were operated on, according to a standardised protocol. More than half the patients were treated with a dynamic hip screw, more than 30% with a cemented biarticulated hemiarthroplasty and fewer than 15% with cannulated screws. The mean admission time in the orthopedic department was 18.7 days and was poorly correlated with the type of surgery or with the place to which the patients were discharged. After hospitalisation, most patients admitted from an institution went back to that institution. More than one-third of the patients admitted from their home went back home but over 40% used rehabilitation facilities. After four months, 32 patients had died, 27 were lost to follow-up and six had been reoperated. Of the independent patients, at least 24% were institutionalised and more than 60% lived at home. Although hip fractures in the elderly are expensive and debilitating, adequate operative treatment and rehabilitation can reduce costs by limiting the hospital stay, lowering reoperation rates and by favouring reintegration into their prefracture surroundings. PMID- 12769016 TI - Teardrop-femoral head distance after shelf acetabuloplasty for Perthes' disease. AB - Shelf acetabuloplasty is an accepted treatment for Perthes' disease. Favourable outcomes have been reported. This study aimed at determining whether changes in femoral head subluxation after shelf acetabuloplasty occur and whether they predict outcome. We assumed that reduction of subluxation after shelf acetabuloplasty would improve the long-term outcome. During follow-up, growth presented as a source of error when comparing the amount of absolute subluxation. Correction for growth was achieved by calculating a subluxation index. This index showed a significant reduction during follow-up. There was no correlation between clinical and radiological outcome, but the fact that the subluxation index showed a decreasing trend could be promising regarding outcome. This might be one of the reasons why shelf acetabuloplasty produces its effect. Following shelf acetabuloplasty the enlarged acetabulum could possibly direct the growth of the femoral head to a better-contained joint. This could result in better joint congruity and lesser tendency to degenerative arthritis. PMID- 12769017 TI - Posterior cruciate ligament avulsion from the tibia: fixation by a posteromedial approach. AB - The authors present their experience with a posteromedial approach for fixation of posterior cruciate ligament avulsion from the tibia. The approach is easy, safe and demands no great technical prowess or instruments. Some minor modifications and technical tips for a safer exposure and a better fixation are highlighted. This is a reproducible method for achieving good stability in these avulsion fractures, where early intervention prevents significant late disability. Even in cases delayed up to 3 months we advocate fixation after gradual intra-operative traction and multiple longitudinal stab incidisons, if the ligament with the attached bony fragment has retracted proximally. A single 4 mm screw gives sufficient initial stabilisation to allow supervised mobilisation. A high index of suspicion should be maintained in all dash-board injuries presenting with femoral shaft fractures, especially when the patella is also fractured. The diagnosis may be missed in the acute setting if the bony avulsion is not adequately appreciated; routine MRI in this situation is a good option. PMID- 12769018 TI - Revision arthroscopic partial meniscectomy of the knee. A retrospective review. AB - The aim in this study was to find out if there were any revision operations in patients who underwent an arthroscopic partial meniscectomy and we attempted to identify the factor(s) that may be associated with the need for that revision. We reviewed 1,603 patients who underwent an arthroscopic partial meniscectomy, and 16 cases of revision were identified (rate 1%). Nine factors were analysed: age, sex, affected knee, affected meniscus, private or National Health Service (NHS) patient, symptoms prior to the most recent meniscectomy, type of tear, history of re-injury and the progression of Outerbridge changes to the articular surfaces. There was a significant risk for a revision meniscectomy in patients with horizontal/partial thickness tears compared to flap tears. No additional factor analysed was significantly associated with a revision procedure. PMID- 12769019 TI - The use of biodegradables in the treatment of osteochondritis dissecans of the knee: fiction or future? AB - The use of biodegradable fixation devices in the operative treatment of osteochondritis dissecans of the knee could avoid a second operation for removal of the hardware, but what are the disadvantages? Seven osteochondritis dissecans lesions, non-displaced in four adult knees and in one adolescent knee and displaced in two knees of adolescents, were treated by drilling and stabilisation with biodegradable pins, resulting in primary consolidation in the five non displaced lesions and failure in the two detached lesions. However, two detached fragments in adults, primarily fixed with one metallic compression screw and three biodegradable pins both consolidated. In another adult patient, the fixation with two compression screws failed. A study of the available literature and the results of our limited experience seem to indicate that the primary operative treatment of choice of a non-detached osteochondritis dissecans lesion is drilling and fixation with biodegradable pins. However, if this regimen fails or in patients with a detached lesion, one metallic screw and a few additional biodegradable pins appear to constitute the best method of fixation. The use of biodegradable screws is still hazardous, because of the long degradation time and subsequent risk of erosion of the opposite cartilage and tissue reaction. Other resurfacing options are available for failures or fragmented or non-vital lesions. PMID- 12769020 TI - Alveolar soft part sarcoma--reciprocal translocation between chromosome 17q25 and Xp11. Report of a case with metastases at presentation and review of the literature. AB - The molecular pathogenesis of alveolar soft part sarcoma, a rare tumor with uncertain histogenesis, was elucidated recently and was shown to be due to a translocation between chromosome 17q25 and Xp11 resulting in a fusion product between TFE3 (a transcription factor gene) at chromosome Xp11 and a novel gene designated as ASPL at chromosome 17q25. This results in the transcriptional dysregulation in the pathogenesis of this neoplasm. Of the 12 cases reported so far, the translocation was due to non-reciprocal translocation in 11 cases with only one case demonstrating a reciprocal translocation with respective fusion products. We report yet another case with reciprocal translocation between chromosomes 17q25 and Xp11 with TFE3/ASPL fusion product who presented with metastatic disease. A standard cytogenetic analysis of primary tumor cells with G banding revealed an abnormal karyotype: 46, X, t(X;17)(p11;q25)[15]/46,XX[5]. PCR analysis of the frozen tumor tissue revealed a type 1 fusion product as described in the literature. We demonstrate a rare cytogenetic abnormality in ASPS, namely reciprocal translocation between chromosomes 17q25 and Xp11 with demonstration of molecular fusion product between TFE3 and ASPL in a patient who initially presented with pulmonary metastases. PMID- 12769021 TI - Posterior sternoclavicular dislocation. Report of two cases. AB - Posterior sternoclavicular dislocation is a relatively uncommon lesion, but must always be considered in the event of trauma to the scapulothoracic area in which initial radiology does not show signs of fracture. Its diagnosis and treatment must be carried out promptly because of the possible serious complications that may occur through the clavicle compressing nearby structures. The authors report two cases, which were diagnosed by CT-scan. In the first case, treatment consisted in orthopedic reduction, while in the second case open reduction and fixation with Kirschner wires was required. The result was satisfactory in both cases, and the patients remain asymptomatic three and five years after trauma. PMID- 12769022 TI - Presumptive infection of a total hip prosthesis by Mycobacterium tuberculosis: a case report. AB - Infection of a total hip prosthesis by Mycobacterium tuberculosis (MT) is an uncommon complication. We report a patient with septic loosening of a total hip prosthesis with a presumptive diagnosis of MT infection. His previous history was negative to any form of tuberculosis, and there was no evidence of a primary focus on the radiological study of the chest. He was diagnosed initially as presenting aseptic loosening and revision surgery was performed. Six months after surgery, septic loosening was diagnosed. The prosthesis was then removed. The histological study showed caseum granulomas and acid-fast bacilli, and routine cultures of the synovial fluid were negative. Since only a positive Lowenstein culture would have confirmed the diagnosis with certainty, a presumptive diagnosis of MR infection was made based on the findings in the histological study. Tuberculostatic treatment was administered for 9 months. At six years follow-up no signs of infection are present, neither by clinical or radiological criteria. PMID- 12769023 TI - Anterior elbow dislocation with recurrent instability. AB - Anterior elbow dislocations are often associated with severe soft tissue lesions. The mechanism of dislocation is usually an anteriorly directed blow on the proximal ulna after a fall on a flexed elbow. Simple anterior elbow dislocations are traditionally treated by closed reduction followed by a short period of immobilisation. Fracture-dislocations are usually unstable after reduction and necessitate surgical management. The authors report a case of acute anterior dislocation with a concomitant radial head fracture that was complicated by recurrent instability. The literature is reviewed to discuss a treatment strategy. PMID- 12769024 TI - Bilateral simultaneous combined intra- and extracapsular femoral neck fracture secondary to nutritional osteomalacia: a case report. AB - A case of bilateral simultaneous combined extra- and intracapsular femoral neck fractures is presented in an adult Asian male patient suffering from nutritional osteomalacia. The radiological and biochemical findings were classical for osteomalacia with Looser's zones affecting the shoulder and the pelvic girdle. Treatment consisted of bed rest and calcium and vitamin-D replacement. The fractures healed without surgery. A high index of suspicion for nutritional osteomalacia among Asian immigrants especially among the low socio-economic classes is required in the Western community. Early diagnosis and treatment prevents chronic disability and skeletal deformity. PMID- 12769025 TI - Fracture of an elastic humeral nail. A case report. AB - The use of elastic intramedullary nails for the treatment of diaphyseal fractures of the humerus has been associated with few complications. We report a fracture of an elastic Marchetti-Vicenzi nail used to treat a pathological fracture of the humerus. The failure of the intramedullary nail followed low-energy trauma and was presumably the end result of metal fatigue, as the original fracture had only healed after 13 months. PMID- 12769026 TI - Overcoming technology mistakes. AB - You do not always need to purchase the very best technology for every situation. Dentists spend a great deal of time trying to find out from gurus and experts which technology is the best. I personally believe that it is far more important to acquire technologies that will integrate into your office properly, provide you with a solid return on investment, and make you productive and profitable rather than simply acquiring the best technologies available. The best method for achieving this objective is the development and implementation of a well-thought out and properly researched technology plan. PMID- 12769027 TI - Topically applied minocycline microspheres: why it works. AB - This article presents the results of a single-arm, open-label, multicenter clinical trial of the topical use of sustained-release minocycline hydrochloride (HCl) microspheres as an adjunct to scaling and root planing. The objective of this study was to evaluate the long-term safety and efficacy of the subgingival application of resorbable minocycline microspheres as an adjunct to scaling and root planing in the treatment of chronic periodontitis. The primary outcome measures were the reduction in probing pocket depth at 9- and 12-month evaluations, and the percent of bleeding upon probing. A total of 173 patients with moderate-to-severe chronic periodontitis were enrolled in this multicenter clinical trial. All patients received full-mouth scaling and root planing plus minocycline microspheres in all periodontal pockets that probed > or = 5 mm. All sites treated at baseline and any new sites > or = 5 mm again received minocycline microspheres at 3- and 6-month follow-up appointments with no further scaling and root planing. Significant improvements in all clinical parameters measured were found at all time points (1, 3, 6, 9 and 12 months). The product was found to be well-tolerated by patients, safe, and easy to deliver. Scaling and root planing with the topical application of minocycline microspheres appeared to give better results than would have been expected with scaling and root planing alone. PMID- 12769028 TI - Digital photography. PMID- 12769029 TI - Rehabilitation of endodontically treated teeth using the radiopaque fiber post. AB - Metallic posts fall short of satisfying contemporary guidelines for ideal post/core rehabilitation. Along with technological improvements in adhesive resins, cements, and composite restoratives, the evolution of fiber-reinforced posts allows the rehabilitation of endodontically treated teeth with greater esthetics and virtually no predisposition to root fracture. At least one fiber post system now complies with all of the ideal post characteristics described in the endodontic text. This article describes the potential for displacement of metal posts by low-modulus fiber posts, the differences between them, and the development and placement of a radiopaque, translucent, double-tapered fiber post. PMID- 12769030 TI - A new cement system for the general/cosmetic dental practice. PMID- 12769031 TI - The relationship between periodontic and endodontic diseases. PMID- 12769032 TI - Credibility: the future of dentistry depends on it. PMID- 12769033 TI - Diet and wellness: why the no-diet plan is your best plan. PMID- 12769034 TI - Bad news for dieters: a focus on ephedrine. PMID- 12769035 TI - Precepts of palliative care. PMID- 12769037 TI - [En bloc ligation of the thoracic duct in twelve dogs with idiopathic chylothorax]. AB - Twelve dogs with idiopathic chylothorax were treated by en bloc ligation of the thoracic duct. Six dogs recovered completely. Minimal follow-up in this group was 12 months. Five dogs were euthanized within 5 months of surgery because of persisting liquothorax: two with chylous and three with non-chylous effusions. One dog was euthanized 2 months after surgery because of a recurrence of dyspnoea. It is concluded that the results are comparable with those of a previously described surgical treatment; however, the technique described here is less complicated because it does not require mesenteric lymphangiography. PMID- 12769038 TI - [Alcohol poisoning in dogs]. AB - The clinical symptoms of ethanol intoxication in a Jack Russell terrier dog of 5 kg, that consumed 50 ml of whisky (40%) within 15 minutes on an empty stomach are described. The clinical case is completed with literature information on clinic, pathogeneses and therapy. PMID- 12769039 TI - [Communication in the case of a mouth affection...]. PMID- 12769040 TI - [Babesiosis]. PMID- 12769041 TI - [A new product in the vaccination gap: the vector vaccine. Merial introduced an innovative influenza-tetanus vaccine for horses]. PMID- 12769042 TI - [Animal euthanasia. Medical and ethical aspects]. PMID- 12769043 TI - Parental, family, and home characteristics associated with cigarette smoking among adolescents. AB - PURPOSE: This study examines the relationship between smoking-related parental, family, and home factors with adolescents' cigarette use. DESIGN: Cross-sectional surveys of adolescents, via a self-administered questionnaire in classrooms, and their parents, via a telephone interview, were conducted. SETTING: Fifteen suburban and rural communities within Minnesota. SUBJECTS: The study sample included 8th, 9th, and 10th grade public school students and their parents. MEASURES: The dependent variable was monthly tobacco use among students. The independent measures were parental, family, and home smoking-related characteristics. There were 1343 parent-child dyads with completed surveys. RESULTS: The final, multivariate logistic regression model found the following variables to be independently related to higher smoking rates among adolescents: child's grade (odds ratio [OR] = 3.03 for 10th vs. 8th), parents' permissiveness of adult smoking (OR = 1.80), parents' having higher normative estimates of how many people smoke (OR = 1.70), parents' decreased likelihood of punishing their teenager for smoking (OR = 1.65), smoking by an adult living in the home (OR = 1.99), and sibling smoking (OR = 8.95). Lack of communication about consequences for breaking family smoking rules was associated with lower smoking rates among adolescents (OR = .49). CONCLUSION: The results of this study highlight the importance of parental smoking norms and attitudes and smoking role models in the home. It is important that smoking prevention strategies target and include the entire family. Limitations of the study are its cross-sectional design and that the sample was primarily white. PMID- 12769044 TI - Impediment profiling for smoking cessation: results of a pilot study. AB - OBJECTIVE: To test the effectiveness of a smoking cessation program based on "impediment profiling," the elucidation of an individual participant's personal barriers, with provision of tailored interventions accordingly. METHODS: A literature search was conducted to identify established impediments to smoking cessation. A long impediment profiler (LIP) was developed from validated survey instruments and used as a screening tool to identify individuals' barriers to quitting. Once barriers were identified, participants were assigned to up to seven interventions. Self-reported smoking cessation was confirmed with measurements of carbon monoxide concentrations in expired air of < or = 10 ppm. RESULTS: Nineteen adults participated in the pilot program. At the year 1 mark, 63.2% of the study population was smoke-free. The mean number of impediments of the study population was 3.5 +/- 1.5. There was a negative association between subjects' quit status and the following impediments: stress (p = .0061), anxiety (p = .0445), and depression (p < .001). No single impediment was predictive of quit status. CONCLUSIONS: Impediment profiling as a basis for tailored smoking cessation intervention is associated with a high quit rate in this initial study, and it appears promising. Long-term follow-up is warranted, as is replication in a larger cohort with a concurrent control group. PMID- 12769045 TI - Applying the transtheoretical model to pregnancy and STD prevention: a review of the literature. AB - PURPOSE: To conduct a systematic review of the peer-reviewed literature on the Transtheoretical Model (TTM) and pregnancy and STD prevention. DATA SOURCES: Computer database search (Applied Social Science Index and Abstracts [ASSIA], Biological Abstracts, Criminal Justice Abstracts, CINAHL-Allied Health, Current Contents, Current Index to Journals in Education, Education Index, ERIC, Excerpta Medica, Family Index, Index Medicus, Medline, Multicultural Education Abstracts, PsychInfo, Psychological Abstracts, Research Alert, Social Science Citation Index, Social Work Abstracts, and Sociological Abstracts), and manual journal search. STUDY INCLUSION AND EXCLUSION CRITERIA: All English, peer-reviewed, original articles on the TTM as it relates to pregnancy and STD prevention published prior to December 31, 2001, were included. Editorials, commentaries, thesis/dissertations, unpublished studies, technical reports, and books were not included. DATA EXTRACTION METHODS: Articles were categorized as Intervention, Population (stage distribution), or Validation studies. Within each category, articles were subdivided into groups, summarized, and analyzed. DATA SYNTHESIS: The 32 articles reviewed included 9 intervention studies, 11 population studies, and 12 validation studies. Studies represented a variety of U.S. populations of a broad demographic range. Evidence both for and against criterion-related and construct validity of the TTM was found. MAJOR CONCLUSIONS: Age, partner type, gender, reasons for engaging in safer sex behaviors (i.e., pregnancy vs. disease prevention), self-efficacy, sexual assertiveness, and perceived advantages and disadvantages of condom use were related to stage of change. The use of the TTM to reduce risk of pregnancy and STDs is a relatively new, but important, area of research. However, because of the wide-ranging differences in methodologies and samples, no strong conclusions about its effectiveness can yet be made. PMID- 12769046 TI - Congruence of readiness to change, self-efficacy, and decisional balance for physical activity and dietary fat reduction. AB - PURPOSE: The current study examines the relations between decisional balance and self-efficacy variables on stage of change between the behaviors of avoiding dietary fat and increasing exercise. DESIGN: A cross-sectional design was used. SETTING: The current study took place in public primary care clinics from four sites across Louisiana. Clinics were associated with teaching hospitals and located in urban and rural areas. SUBJECTS: Subjects included 515 adult outpatients, 60% African-American, 81% women, and 43% married. The age ranged from 18 to 87 years old, and the mean age was 45 (SD = 14). Patients were predominantly low-income (mean household income of $490 per month) and uninsured (71%). MEASURES: Standard questionnaires were given to asses stage of change, decisional balance, and self-efficacy for exercise and dietary fat reduction. RESULTS: Although the chi 2 analysis revealed that dietary fat and exercise stage of change were significantly related, Pearson chi 2 (df = 16) = 74.30, p < .001, 35% of the sample was stage incongruent between behaviors (e.g., a significant percentage of exercise maintainers were precontemplators for reducing dietary fat). Only 27% of the sample was in the same stage for both behaviors. Correlations and multivariate analyses of variance (MANOVAs) indicated that relationships between behaviors were similar to those found previously within behaviors; however, the effect sizes were markedly attenuated. CONCLUSIONS: These results have implications for healthcare providers working with weight management. Accurate assessment of readiness for change for both exercise and dietary fat consumption is critical. For many patients, readiness for change differs dramatically between the two behaviors, and interventions may need to be tailored more precisely. Providers may need to use more active, behaviorally focused interventions for the more advanced behavior while simultaneously implementing more cognitively focused interventions for the less advanced one. PMID- 12769047 TI - Organizational health promotion: broadening the horizon of workplace health promotion. AB - This article argues that efforts to improve the health and well-being of the workforce should begin with the organization itself. The term organizational health promotion is introduced to expand the scope of worksite health promotion. Organizational health promotion delves into the basic structural and organizational fabric of the enterprise--to how work is organized. The core themes of healthy work organization are introduced, and the status of our ability to identify organizational risk factors is discussed. A conceptual model of healthy work organization is presented, along with a process for expanding the health promotive capacity of the organization. The final section addresses challenges related to adopting an organizational health promotion perspective. PMID- 12769048 TI - Millennial heresy. AB - Beginning in 1996, protease inhibitors and three-drug combination therapy revolutionized HIV treatment. At least in the developed world, a new highly aggressive treatment standard emerged almost overnight. Not surprisingly, four years of experience with the new therapies have shown a few cracks in that standard and it may be time to reconsider some of the common beliefs about therapy. PMID- 12769049 TI - Nelfinavir potency concerns. AB - Results from several recent studies may help clarify how to best use anti-HIV drugs as part of first line therapy. These results seem to suggest that nelfinavir (Viracept), the most widely used protease inhibitor, may not be as potent as other drugs in its class or some of the non-nucleoside RT inhibitors (NNRTIs). This article provides an overview of the studies. PMID- 12769050 TI - Anti-HIV drug update. AB - Slow but steady progress continues on a number of new anti-HIV drugs in the three drug classes. If approved, some of these represent true second-generation therapies that will offer some degree of advance over current drugs. Others are not that much different from today's drugs. The article below combines information from the most recently announced studies, including those presented at the 7th Conference on Retroviruses and Opportunistic Infections. PMID- 12769051 TI - Manufacturer end adefovir development. AB - Gilead Sciences has stopped development of its drug, adefovir, for use against HIV infection. This news came after the Food and Drug Administration (FDA) informed Gilead that adefovir would not be approved based on existing data and that additional studies would be necessary if it wants the FDA to reconsider the matter. The Antiviral Drugs Advisory Committee to the FDA had earlier recommended that the drug not be approved on the grounds that its potential serious side effects outweighed its potential benefits. PMID- 12769052 TI - New drugs in development. AB - In the last issue of PI Perspective (#28), we reviewed some new anti-HIV drugs in development. Since then, setbacks have occurred that will delay, and in some cases stop, the development of some of these therapies. However, other therapies show promising activity and continue through the development process quickly. PMID- 12769053 TI - A case of brain damage due to a high-energy proton beam. PMID- 12769054 TI - Antecedents and consequences of work-family conflict: a prospective cohort study. AB - This study examined both risk factors for the onset of work-family conflict and consequences in terms of need for recovery and prolonged fatigue for men and women separately. Two-year follow-up data from the Maastricht Cohort Study on "Fatigue at Work" (n = 12,095) were used. At baseline, the prevalence of work family conflict was 10.8% (9.0% in women; 11.1% in men), the cumulative incidence at 1 year follow-up was 5.1%. For men, several work-related demands, shift work, job insecurity, conflicts with coworkers or supervisor, having full responsibility for housekeeping, and having to care for a chronically ill child or other family member at home were risk factors for the onset of work-family conflict, whereas decision latitude and coworker and supervisor social support protected against work-family conflict. In women, physical demands, overtime work, commuting time to work, and having dependent children were risk factors for work-family conflict, whereas domestic help protected against work-family conflict at 1 year follow-up. Work-family conflict was further shown to be a strong risk factor for the onset of elevated need for recovery from work and fatigue. PMID- 12769055 TI - Implantation of a biventricular pacing system in a patient with a coronary sinus not communicating with the right atrium. AB - Intubation of the coronary sinus failed during implantation of a biventricular pacing system. An angiogram of the left coronary artery showed in the venous phase a coronary sinus not communicating with the right atrium. The coronary sinus was draining into a persistent left superior vena cava communicating with the left subclavian vein. The coronary sinus lead was successfully implanted through the persistent left superior vena cava, whereas the atrial and ventricular leads were implanted through the right superior vena cava in a conventional way. PMID- 12769056 TI - [On a day like no other: Ophthalmological Academy in Dusseldorf]. PMID- 12769058 TI - [Federal tax liability of medical procedures?]. PMID- 12769057 TI - [Introduction of a quality management system in the physician's practice]. PMID- 12769059 TI - Use and abuse of new technologies in colorectal surgery. PMID- 12769061 TI - Anal sphincter replacement: the solution to a problem? PMID- 12769060 TI - [Symbolic language of the eye in mythology and history]. PMID- 12769062 TI - Manometric variables in rectocele patients with symptomatic constipation. PMID- 12769064 TI - Proceedings of the symposium on mucus hypersecretion in respiratory disease. London, United Kingdom, 26-28 February 2002. PMID- 12769063 TI - Treatment of rectovaginal fistulas. PMID- 12769067 TI - [Reconstruction of the external ear after total amputation]. PMID- 12769068 TI - [We should pay more attention on clinical research]. PMID- 12769069 TI - [Diagnosis and treatment for subarachnoid hemorrhage patients]. PMID- 12769071 TI - Proceedings of the 54th International Symposium on Crop Protection. Gent, May 7, 2002. PMID- 12769070 TI - [Reconstruction of the ear or ear region in skin avulsion or burns]. PMID- 12769072 TI - Regulation of Carbon Metabolism: sensing, signalling and partitioning. Proceedings of a symposium. Swansea, United Kingdom, 8-12 April 2002. PMID- 12769073 TI - Application of Molecular Markers in Studies on Plants. September 25-29, 2002. Warsaw, Poland. Symposium proceedings. PMID- 12769074 TI - Endogenous microbial contamination of cultured autologous preparations in trials of cancer immunotherapy. AB - BACKGROUND: Standardization of the manufacturing and processing of cellular immunotherapy products is necessary to ensure patient safety, establish efficacy, and demonstrate potency. Recognition of the autologous donor as a likely source of microbial contamination of cellular immunotherapy products may improve patient care and reduce expense to the laboratory. METHODS: Data on 243 immunotherapy products manufactured in the authors' institution between December 1997 and June 2001 were retrospectively reviewed. Also reviewed were the case reports of four patients whose autologous immunotherapy products were contaminated. RESULTS: Twenty-five (10%) of the 243 immunotherapy products processed were positive on one or more tests for microbial contamination. In six (24%) of the products, the source of microbial contamination was the autologous donor. In 17 of the remaining 19 products, test results were judged to be false-positive. DISCUSSION: The unique processing techniques and stringent controls involved in the manufacture of cellular immunotherapy products may result in changes in the sources of microbial contamination routinely encountered. The identification of the autologous donor as a potential source of the microbial contamination of the product may assist the clinician and the laboratory in troubleshooting products with positive results on microbial sterility testing. Also, the number of false positive results in this study indicates that further research is needed to maximize the specificity of testing while maintaining the present high sensitivity. PMID- 12769076 TI - Proceedings of the 2nd International Symposium on Cytokines and Depression. Dinard, France, May 12-14, 2001. PMID- 12769075 TI - [Severe hip dislocations in patients with infantile cerebral palsy--is surgical reconstruction sensible?]. PMID- 12769077 TI - [Roentgen morphologic evaluation of tendinosis calcarea of the shoulder is interobserver judgment dependent]. PMID- 12769078 TI - [Progress on pathogen and onset mechanism of primary pulmonary hypertension]. PMID- 12769079 TI - [Social medicine consequences for total endoprosthesis patients. Postoperative loadbearing of hip and knee prostheses]. PMID- 12769080 TI - [Non-invasive quantification of cartilage thickness and cartilage damage with MRI]. PMID- 12769081 TI - [Quality management according to DIN ISO 9001:2000--key requirements and applications in rehabilitation]. PMID- 12769082 TI - [Proceedings of the 10th Conference of Psychiatric Treatment. Gdansk, Poland, June 2002]. PMID- 12769083 TI - [Disease, experiment and psychology in mid-19th century France]. AB - This article offers a contribution to the thesis that considers H. Taine and Th. Ribot the theorists of the "pathological method" in psychology, and thus the forefathers of French experimental psychology. At the same time it points out that the idea of equivalence between observation of the pathological and experimentation could already be found in th psychological medicine of the 1840s. The author examines the early years of the Annals medico-psychologiques (1843 1844), the constitution of the Societe medico-psychologique (1852), and the work of J. Moreau de Tours (1843) on hashish and mental alienation, with the intention of demonstrating that in the 1850s French psychiatry was actively engaged in the debate on the Science des rapports du physique et du moral, and that consequently the organicistic paradigm, proclaimed in those years in Germany, had not yet spread to France. PMID- 12769085 TI - [Proceedings of the Vienna Radiology Symposium 2002]. PMID- 12769084 TI - The state of the medigap market in Massachusetts. PMID- 12769086 TI - Altruism--measuring the non-existence? PMID- 12769087 TI - Structured interactive sessions: a cooperative-learning technique. PMID- 12769088 TI - Medicare in Massachusetts: the impact of the 1997 Balanced Budget agreement. PMID- 12769089 TI - Graduate entry to medical studies: thoughts fron 'down under'. PMID- 12769090 TI - Will alcohol be the next tobacco? PMID- 12769091 TI - The cost and availability of prescription drugs--what can we do in Massachusetts? PMID- 12769092 TI - Abstracts of the 2003 Research Society on Alcoholism and the 12th Congress of the International Society for Biomedical Research on Alcoholism. June 21-25, 2003. Fort Lauderdale, Florida, USA. PMID- 12769093 TI - [XXXVII Congress of the French College of Vascular Pathology. Paris, France, 11 14 March 2003. Abstracts]. PMID- 12769094 TI - WHO is worried that China is under-reporting SARS. PMID- 12769095 TI - Cultural competence and health care in Massachusetts: where are we? Where should we be? PMID- 12769097 TI - [The Neurological Surgery: table of contents for volume 1-30, 1973-2002]. PMID- 12769096 TI - Mental health care in Massachusetts. PMID- 12769099 TI - [The 46th annual meeting of the Japanese Society of Nephrology. Tokyo, Japan. May 22-24, 2003. Abstracts]. PMID- 12769098 TI - [Christian Gottfried Nees von Esenbeck: correspondence with Johann Wolfgang von Goethe]. PMID- 12769100 TI - [The 104th Congress of the Japanese Society of Otolaryngology. Tokyo, Japan. May 22-24, 2003. Abstracts]. PMID- 12769102 TI - Bovine tuberculosis: still a world health problem. PMID- 12769101 TI - [A new measurement of pharmaceutical prescriptions of the Ebers papyrus]. AB - Starting point of this article is a problem obvious to anybody who reads the medical papyri of ancient Egypt from the New Kingdom and the Late Period: the prescriptions contain two dyadic systems of notation for measures of capacity without specifying the unit of measurement, firstly the Horus eye fractions 1/2, 1/4, 1/8, 1/16, 1/32 and 1/64 which are unanimously assumed to refer to the single Heqat and secondly the, normal' dyadic fractions from 1/2 to 1/128, which have been interpreted in widely different ways, as fractions of the Henu-, the Ro , the single Heqat-, the 5-Ro- or the Dja-measure, respectively. Interpretations suggested in translations and commentaries have remained inconclusive and unsatisfactory. PMID- 12769104 TI - Genetic testing: policy issues for the new millennium. AB - The purpose of this article is to provide a brief overview of the policy considerations underlying the Organization for Economic Cooperation and Development (OECD) workshop, "Genetic Testing: Policy Issues for the New Millennium," which was organised and hosted by the governments of the United Kingdom (UK) and Austria in Vienna, 23-25 February 2000, with the support of the European Commission (EC). It reviews the various topics presented at the meeting and offers a brief summary of its main conclusions. PMID- 12769103 TI - [Primary biliary cirrhosis. Primary sclerosing cholangitis]. PMID- 12769105 TI - Pregnant embodiment and women's autonomy rights in law: an analysis of the language and politics of Winnipeg Child and Family Services v. D.F.G. PMID- 12769106 TI - On-line or on-call? Legal and ethical challenges emerging in cybermedicine. PMID- 12769107 TI - Worst choice: why we'd be better off without Roe. PMID- 12769110 TI - The right to die: death with dignity in America. PMID- 12769108 TI - Physician-assisted suicide: new protocol for a rightful death. PMID- 12769111 TI - Recognizing the need for uniform international regulation of developing biotechnology: a focus on genetic experimentation. PMID- 12769112 TI - Biotechnology and the new property regime in human bodies and body parts. PMID- 12769113 TI - Privacy on thin ice? Considering the California Court of Appeal decision in Johnson v. Superior Court. AB - In Johnson v. Superior Court, the California Court of Appeal determined that a provision of a contract limiting the discovery of the identity of a sperm donor was against public policy and that the privacy interest did not protect against disclosure of this information. Although the court's analysis of the public policy exception to the enforcement of contracts was unnecessary, the opinion properly balances California's and petitioners' interests against an anonymous donor's privacy right. PMID- 12769114 TI - Reconceptualizing informed consent in an era of health care cost containment. PMID- 12769115 TI - To patent or not to patent: the European Union's new biotech directive. PMID- 12769116 TI - Senior prescription drug coverage: the state of the medigap markets in Massachusetts and New Hampshire. PMID- 12769117 TI - Security interests in human materials. PMID- 12769118 TI - Heriberto Duarte Sr. v. Chino Community Hospital. PMID- 12769119 TI - Gestational surrogacy contract. PMID- 12769120 TI - The law of the sacred cow: sacrificing the First Amendment to defend abortion on demand. PMID- 12769121 TI - International protection of genetic information: the progression of the Human Genome Project and the current framework of human rights doctrines. PMID- 12769122 TI - Disputing over embryos: of contracts and consents. PMID- 12769123 TI - Female-selective abortion in Asia: patterns, policies, and debates. AB - Since the early 1980s, the use of sex-selective abortion increased in many Asian contexts. Estimates indicate that several million female fetuses were aborted in the last two decades of the twentieth century. This article takes a currently unusual approach for a cultural anthropologist in pursuing cross-national comparisons of trends in sex-selective abortion. The risks involved in such an approach are taken in the hope that it will yield insights not gained through localized analysis. After reviewing the available evidence on female-selective abortion, I discuss features of Asian culture that support strong son preference. Next I review the related issues of increased technological availability for prenatal sex selection and national policies about sex selection. Last, I consider several positions on female-selective abortion and how cultural anthropology may contribute to understanding the global context and consequences of prenatal gender discrimination. PMID- 12769124 TI - Preventive surgery: the impact of molecular biology in surgery. PMID- 12769125 TI - An analysis of initiative petition 99-4, "An Act to Protect the Rights of Patients and to Promote Access to Quality Health Care for All Residents of the Commonwealth". PMID- 12769126 TI - Current awareness on yeast. PMID- 12769127 TI - Rosuvastatin: a highly efficacious statin for the treatment of dyslipidaemia. AB - Rosuvastatin is a synthetic enantiomer that is hepatoselective, relatively hydrophilic and has minimal metabolism via the cytochrome P450 3A4 system (similar to pravastatin). Rosuvastatin, like atorvastatin, has a plasma half-life of about 20 h and is a potent inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase. The majority of HMG-CoA reductase inhibitory activity in plasma is associated with the parent rosuvastatin compound. In a Phase II study, rosuvastatin across a dose range of 1 - 80 mg lowered low-density lipoprotein cholesterol (LDL-C) by 34 - 65%. Phase III trials have demonstrated greater reductions in LDL-C for rosuvastatin compared to atorvastatin as well as greater increases in high-density lipoprotein cholesterol (HDL-C). The drug appears to be well tolerated at all doses up to 80 mg/day. A starting dose of 10 mg will reduce LDL-C by approximately 50%, which should adequately treat most patients to within the National Cholesterol Education Program (NCEP) Adult Treatment Panel III (ATP III) goals. PMID- 12769128 TI - The value of a Waters' projection. PMID- 12769129 TI - Sphenoid sinus mucocele. PMID- 12769130 TI - Reconsidering the 10(5) rule. PMID- 12769131 TI - Keeping their feet on the ground. PMID- 12769132 TI - [Diabetes, Conn syndrome and osteoporosis. What is new in diagnosis and therapy?]. PMID- 12769133 TI - Cognitive aspects of survey methodology and quality of life assessment. PMID- 12769134 TI - Cognitive psychology and self-reports: models and methods. AB - This article describes the models and methods that cognitive psychologists and survey researchers use to evaluate and experimentally test cognitive issues in questionnaire design and subsequently improve self-report instruments. These models and methods assess the cognitive processes underlying how respondents comprehend and generate answers to self-report questions. Cognitive processing models are briefly described. Non-experimental methods--expert cognitive review, cognitive task analysis, focus groups, and cognitive interviews--are described. Examples are provided of how these methods were effectively used to identify cognitive self-report issues. Experimental methods--cognitive laboratory experiments, field tests, and experiments embedded in field surveys--are described. Examples are provided of: (a) how laboratory experiments were designed to test the capability and accuracy of respondents in performing the cognitive tasks required to answer self-report questions, (b) how a field experiment was conducted in which a cognitively designed questionnaire was effectively tested against the original questionnaire, and (c) how a cognitive experiment embedded in a field survey was conducted to test cognitive predictions. PMID- 12769135 TI - Pretesting survey instruments: an overview of cognitive methods. AB - This article puts forward the case that survey questionnaires, which are a type of measuring instrument, can and should be tested to ensure they meet their purpose. Traditionally survey researchers have been pre-occupied with 'standardising' data collection instruments and procedures such as question wording and have assumed that experience in questionnaire design, coupled with pilot testing of questionnaires, will then ensure valid and reliable results. However, implicit in the notion of standardisation are the assumptions that respondents are able to understand the questions being asked, that questions are understood in the same way by all respondents, and that respondents are willing and able to answer such questions. The development of cognitive question testing methods has provided social researchers with a number of theories and tools to test these assumptions, and to develop better survey instruments and questionnaires. This paper describes some of these theories and tools, and argues that cognitive testing should be a standard part of the development process of any survey instrument. PMID- 12769136 TI - Hi! How are you? Response shift, implicit theories and differing epistemologies. AB - Measures of Health Related Quality of Life (HRQL) occupy a continuum, from highly standardized econometric methods such as the time tradeoff and standard gamble to individualized global measures. Each has its vocal adherents, each involves different assumptions about the nature and interpretation of HRQL, and each has potential advantages and disadvantages. In this paper, I begin by exploring two theories which attempt to explain how people make assessments of health over time: 'response shift' and the 'implicit theory of change' model. I show that the theories, which are based on different views of the underlying cognitive processes, make opposite predictions about the validity of prospective and retrospective judgments. I examine the broader issue of individualized vs. standardized questions, and discuss a fundamental epistemological difference which places the current discussion in a broader philosophical context. I propose that a partial resolution may arise from a more careful consideration of the goals of HRQL assessment in a particular situation. PMID- 12769137 TI - Cognitive approaches to summary measurement: its application to the measurement of diversity in health-related quality of life assessments. AB - This paper describes elements of a cognitive approach to summary measurement, as it is applied to health-related quality of life (HRQOL) assessment. A summary measure, by its very nature, represents, but does not reproduce, some body of information. As a consequence, how a summary measure is formulated becomes a critical determinant of its usefulness. In most cases, formal (arithmetic or investigator-based) methods are used. However, it was also noted that a person ordinarily summarizes the information they receive. Qualitative analytic techniques (e.g., cognitive interviewing methods) can provide insight into the cognitive processes underlying such summary formation. These cognitive processes, formulated as algorithms, would also be expected to vary as a function of different groups of respondents and settings. Finding a common denominator, a common algorithm, would provide a consensus summary measure amongst diverse groups whose HRQOL is being assessed. PMID- 12769138 TI - The potential synergy between cognitive models and modern psychometric models. AB - Analyses of cognitive aspects of survey methodology (CASM) and psychometric analysis are two methods that are able to complement each other. We use concrete examples to illustrate how psychometric analyses can test hypotheses from CASM. The psychometrics framework recognizes that survey responses are affected by other factors than the concept being assessed, for example by cognitive factors and processes. Such factors are subsumed under the concept of measurement error. Possible sources of measurement error can be tested, e.g. by randomized experiments. A standard way to reduce measurement error is to ask several questions about the same concept and combine the answers into a multi-item scale that is more precise than the individual items. Techniques like structural equation models use the item correlations to assess the magnitude of measurement error and to test the assumptions behind the multi-item scale, e.g. the effect of common response choices and item time frames. A central problem in modern psychometrics is how to model the mapping of the continuous latent variable onto the item response choice categories. This is achieved by threshold models (e.g. item response models and structural equation models for categorical data). These models can, for example, analyze the impact of mode of administration, test whether the items function in the same way for all people (measurement invariance/differential item functioning) and examine the consistency of responses from any single person. Such analyses provide new possibilities for combining psychometrics and cognitive methods. PMID- 12769139 TI - A theory-based method for the evaluation of individual quality of life: the SEIQoL. AB - Few reports about methods of evaluating quality of life (QoL) among the thousands published since medical interest in the subject slowly began nearly 40 years ago are based upon theory. This paper, prepared in response to a request to furnish an exception (Meadows KA. Introduction to an Advanced Seminar: Assessing Health Related Quality of Life. What can the Cognitive Sciences Contribute? Hull University, October 9, 2000) describes the origins of the Schedule for the Evaluation of Individual Quality of Life (SEIQoL). This derives its cognitive aspects from theoretical studies of perception by Egon Brunswik, their extension to Social Judgment Theory (SJT) by Kenneth Hammond and the application of these ideas to QoL by the present authors and their colleagues. PMID- 12769140 TI - Cognitive aspects of survey methodology and quality of life assessment: summary of meeting. PMID- 12769141 TI - Quality of life for adolescents: assessing measurement properties using structural equation modelling. AB - Assessments for quality of life (QOL) of the adolescent have received relatively little attention in the literature. Although there is no consensus on the definition of adolescent QOL and what aspects should be measured, it is generally accepted that QOL is a multidimensional construct. The objective of this study is to determine the measurement properties of the latent factors underlying adolescent QOL based on a second-order confirmatory factor analysis (CFA). A recursive structural equation model (SEM) is then proposed to determine the direction and magnitude of the interdependent effects among the latent factors. The questionnaire used was the Quality of Life Profile-Adolescent Version (QOLPAV). A sample of 363 adolescents was recruited from 20 secondary schools in Perth, Australia. The second-order CFA suggested that adolescent QOL may be measured by five underlying constructs namely social, environment, psychological, health, and opportunities for growth. The interdependent relations among these constructs identified the environment factor as primary, exerting both direct and indirect effects on the other four factors. PMID- 12769142 TI - Health-related quality of life in children assessed by their parents: evaluation of the psychometric properties of the CHQ-PF50 in two German clinical samples. AB - The aim of the study was to evaluate the psychometric properties of the German translation of the Child Health Questionnaire (CHQ). Parents of two clinical samples were asked to rate the quality of life of their children using the German version of the CHQ. Item internal consistency (item-scale correlation) and internal consistency of scales were tested; quartiles and factor analysis were conducted. The results of the German clinical samples were compared with US clinical samples (t-test). The two clinical German sub-groups were compared using multivariate analysis (MANOVA). The psychometric testing of the CHQ showed good results. Internal consistency of the hypothesized scales were all higher than 0.70. The results of the factor analysis confirmed the results of the US norm sample: As expected, the subscales loaded on two factors explaining 57.4% of the total variance. The group comparison supported the discriminative properties of the CHQ. Preliminary psychometric findings support use of the German version of the CHQ. The next step will be the testing of healthy German samples and development of norm scores. PMID- 12769143 TI - The Spanish version of the Child Health and Illness Profile-Adolescent Edition (CHIP-AE). AB - The aim of the study was to obtain a conceptually equivalent Spanish version of the Child Health and Illness Profile-Adolescent Edition (CHIP-AE), and to test its feasibility, reliability and preliminary construct validity. The methodology used for adaptation was forward-back translation, including two focus groups with adolescents and a panel of experts. Reliability and validity were assessed in healthy convenience samples from school settings (n = 417). Three different illness groups (n = 67) were used to examine differences in health status between healthy, acutely ill, chronically ill and mentally ill adolescents. Preliminary construct validity was examined by comparing mean scores for each of the subdomains to determine if they differed in predicted ways according to age, gender and illness group. The majority of items (154 out of 203) were conceptually equivalent to the original version. Some items (46) had to be modified to increase clarity and/or to adapt them for use in Spain and 3 items were considered not applicable. Single construct subdomains achieved alpha coefficients between 0.65 and 0.92, and intraclass correlation coefficients (ICC) between 0.57 and 0.93. The mentally ill group presented the worst scores in most domains. The Spanish CHIP-AE is acceptable for Spanish adolescents and shows adequate metric characteristics, which are similar to those reported in the US version. PMID- 12769144 TI - Sexual function and quality of life in genitourinary medicine (GUM) outpatients and preliminary validation of a self-report questionnaire measure. AB - A cross-sectional questionnaire survey of 216 men and 191 women attending a genitourinary medicine (GUM) clinic was undertaken to explore the relationship between sexual symptoms and quality of sexual life, and to test the psychometric validity of a pilot self-report measure of Sexual Function and Quality of Sexual Life (SFQoSL). Statistical comparisons were made with three reference groups: volunteers attending GUM for psychosexual counselling, outpatients at an Obstetrics and Gynaecology Department, and staff. Exploratory principal components analysis (with varimax rotation) of questionnaire item responses suggested an 11 (in women) and 13 (in men) factor solution, incorporating four multi-item scales. Internal consistency (Cronbach's alpha) of core items was 0.84 in 186 women (19 items) and 0.87 in 210 men (22 items). Construct validity was supported in comparisons with reference groups using one-way analysis of variance and post-hoc Scheffe testing. Overall, 116 (54%) male and 132 (69%) female GUM outpatients had scores indicating sexual dysfunction. Thirty-seven (17%) men reported erectile dysfunction; 54 (28%) women reported vaginal dryness affecting sex; 48 (25%) women reported genital changes affecting sex; 45 (21%) men and 64 (34%) women reported problems reaching orgasm. PMID- 12769145 TI - Quality of life impact and treatment seeking of Chinese women with urinary incontinence. AB - The objective of this study was to investigate the quality of life (QOL) impact of urinary incontinence (UI) and to examine its relationship with treatment seeking in adult Taiwanese women. We conducted a cross-section in-person questionnaire interview of 1608 adult women living in the Taipei area. The characteristics and incontinence status were recorded. A short form incontinence impact questionnaire (IIQ-7) was used to evaluate the QOL impact of UI. Multiple logistic regression analysis was used to assess the determinative factors for treatment seeking. The mean IIQ-7 score of the 205 (12.7%) women who reported urinary leakage more than once per month in the preceding 12 months was 5.0 (range: 0-19), which showed a significant correlation with the severity of incontinence (r = 0.59, p < 0.001). Women with mixed type UI had a higher IIQ-7 score compared to those with stress or urge UI. Fifty-five (26.8%) incontinent women had sought medical help. Treatment seeking was highly related to IIQ-7 scores as 75% of incontinent women with an IIQ-7 score > 10 in contrast to 5% of those with an IIQ-7 score < or = 3 (p < 0.001) had sought medical care. On multiple logistic regression analysis, perceiving UI as a disease and a higher IIQ-7 score were independent factors predicting treatment seeking. We concluded that UI is a common problem that brings substantial QOL impact to Taiwanese women. The IIQ-7 questionnaire may provide a useful measurement to quantitate the degree of QOL impact, which is largely affected by the severity and type of incontinence. Women who perceive UI as a disease and those with a higher degree of QOL impact are more likely to seek medical help. Furthermore, treatment seeking in Taiwanese women with significant UI may be more common than thought as a great majority of women with higher IIQ-7 scores had sought medical help. PMID- 12769146 TI - Sprain of the neck: quality of life and psychological functioning. A 4-year retrospective study. AB - Aim of the study was to analyse quality of life and psychological functioning in patients with sprain of the neck, to analyse the relationship between complaints, quality of life, psychological functioning and personality factors, and to analyse the profile of patients with whiplash associated disorders (WAD), 4 years after trauma. From the University Hospital Groningen 193 patients with the diagnose sprain of the neck filled out a questionnaire. Of this group 100 subjects did not have complaints before the accident and were therefore at risk for the development of complaints as a result of sprain of the neck. Quality of life and psychological functioning were assessed using the RAND-36 and the SCL 90, respectively. Personality was assessed by means of the Dutch Personality Questionnaire. Of the group at risk (56% women and 44% men, mean age: 33.9, SD: 14.6) quality of life was significantly worse in subjects with complaints (mean: 78.4, SD: 15.5) compared to subjects without complaints (mean: 87.5, SD: 8.7). Psychological functioning did not differ significantly between the group with complaints compared to group without complaints. Personality did not differ between the groups. Personality and complaints together were significantly related to quality of life (r: 0.77) and psychological functioning (r: 0.85). No specific profile of WAD patients was found. In conclusion, personality and complaints influence quality of life and psychological functioning to a considerable extent. PMID- 12769147 TI - A 1-year prospective clinical trial of menicon Z (tisilfocon A) rigid gas permeable contact lenses worn on a 30-day continuous wear schedule. AB - PURPOSE: To establish the safety and efficacy of clinical performance of a novel hyper-O2-transmitting rigid gas-permeable (RGP) lens (tisilfocon A) worn continuously for 30 days (29 nights) equivalent to a conventional control hydrogel lens (etafilcon A) worn for 7 days (6 nights). METHODS: The study was a prospective, open-label, 24-center, concurrent cohort-controlled clinical trial. OUTCOME MEASURES: Outcome measures included rates of adverse events, slitlamp findings, length of wear achieved, lens-corrected visual acuity, corneal refractive changes, dryness, and lens deposits. RESULTS: The RGP lens wear test was equivalent or superior to the hydrogel control performance in all categories. Two-thirds (66.5%) of patients wearing the RGP lens achieved continuous wear periods greater than 22 nights; and 60.4% achieved more than 28 nights of continuous use. There was a statistically significant lower rate of adverse events, considered definitely related to lens wear, for the RGP test lens versus the hydrogel control (none vs. 2.2%; P = 0.007, Fisher's exact two-sided test). CONCLUSIONS: Continuous wear of the tisilfocon A (Menicon Z, Menicon Co., Ltd., Nagoya, Japan) RGP lens for up to 30 nights is a safe and equivalent alternative to 7-day (6-night) hydrogel wear. PMID- 12769149 TI - Corneal endothelial morphology results in the Menicon Z 30-day continuous-wear contact lens clinical trial. AB - PURPOSE: This evaluation was conducted as part of a protocol entitled Evaluation of the Menicon Z Rigid Gas Permeable Contact Lens for up to 30 Days Extended Wear. The purpose of the protocol was to compare corneal endothelial morphology changes after wearing rigid gas-permeable (RGP) Menicon Z contact lenses, continuously for up to 30 days, with ACUVUE (Johnson & Johnson Vision Care) hydrogel contact lenses, worn for up to 6 nights of extended wear. METHODS: Sixty patients, who were adapted to RGP daily wear and soft contact lens daily wear, were recruited at two study sites. The thirty subjects who wore RGP daily wear lenses were fitted with the Menicon Z (tisilfocon A, oxygen permeablility [Dk] = 163) RGP contact lens comprised the test group. The control group subjects (n = 30 former users of daily wear soft contact lenses) were fitted with ACUVUE (etafilcon A, Dk = 28) hydrogel contact lenses. After a 2-week adaptation period of daily wear, subjects began extended wear. Endothelial imaging was performed at the two study sites in this multicenter study (University Hospitals of Cleveland/Case Western Reserve University Department of Ophthalmology [CWRU] and The Ohio State University [OSU]). The hydrogel lens group was instructed to wear their lenses for 7 days and 6 nights before discarding the lenses and to sleep with no lenses on the seventh night. The RGP group was permitted to wear the lenses for up to 30 days and 29 consecutive nights before removing the lenses for cleaning and overnight soaking. RESULTS: CWRU had 24 patients (12 soft contact lens and 12 rigid contact lens) and OSU had 21 patients (12 soft contact lens and 9 rigid contact lens) who completed the study and were included in the analyses. Patients who were withdrawn from the study at CWRU included one RGP patient dropped out because of pregnancy; one RGP patient developed vascularized limbal keratitis and discomfort; one could not be fitted with a bitoric RGP; two soft lens patients moved from the area, and one dropped out because of dry eyes. At OSU, four patients dropped out due to discomfort (two in each lens type);one moved from the study area; one decided not to participate soon after the consent visit; one had worries of reduced vision at 6 months; one subject's attitude changed prior to the 6 month visit; and one subject was withdrawn for reasons of poor study schedule compliance. CONCLUSION: The Menicon Z lens wearers in this study did not show significant endothelial cell morphology changes after 1 year of nearly continuous contact lens wear. This finding is particularly important considering the RGP wearers in this study were older by a average of 10 years (mean age approximately 40) and had worn their contact lenses an average of almost 10 years longer than the soft contact lens wearers. PMID- 12769148 TI - Corneal thickness results in the Menicon Z 30-day continuous wear and ACUVUE 7 day extended-wear contact lens clinical trial. AB - PURPOSE: This study was conducted to measure the corneal thickness changes associated with wearing 30-day tisilfocon A (Dk = 163) and 6-day etafilcon A (Dk = 28) contact lenses. The probable cause of the outcomes is discussed. METHODS: Thirty subjects were fitted with Menicon Z rigid gas-permeable (RGP) contact lenses and a control group (n = 30) was fitted with ACUVUE hydrogel lenses, at two study sites. After a 2-week period of daily wear, subjects began prolonged wear. Measurements were performed at the dispensing visit, and then at the 2-week daily wear, the 1-week, 6-month, and 12-month extended- and continuous-wear visits. RESULTS: Repeated measures analysis of covariance was performed on mean corneal thickness. There was a significant increase in corneal thickness in the hydrogel lens extended-wear group compared to the RGP group (P = 0.03). The mean corneal thickness in the ACUVUE group was 2.58% higher at the 2-week daily wear visit, 2.38% higher at the 1-week extended-wear visit, 2.96% higher at the 6 month, and 2.97% higher at the 1-year visit compared to the RGP continuous-wear patients. DISCUSSION: Patients wearing high-Dk RGP contact lenses for continuous wear may have thinner corneas (within the normal thickness range) probably because of minimal hypoxia-induced corneal stromal swelling; RGP-induced reversible corneal epithelial compression; deceased epithelial sloughing; or long term contact lens-induced keratocyte apoptosis. Patients wearing hydrogel contact lenses for extended wear may have thicker corneas because of hypoxia-induced corneal stromal swelling. Additional evidence of corneal hypoxic stress is seen in the higher prevalence of striae and epithelial microcysts in the hydrogel group and greater variability of corneal thickness measurements in the soft contact lens group in our results. PMID- 12769150 TI - The effect of variably tinted spectacle lenses on visual performance in cataract subjects. AB - PURPOSE: A body of clinical and laboratory evidence suggests that tinted spectacle lenses may have an effect on visual performance. The aim of this study was to quantify the effects of spectacle lens tint on the visual performance of 25 subjects with cataracts. METHODS: Cataracts were scored based on best corrected acuity and by comparison with the Lens Opacity Classification System (LOCS III) plates. Visual performance was assessed by measuring contrast sensitivity with and without glare (Morphonome software version 4.0). The effect of gray, brown, yellow, green and purple tinting was evaluated. RESULTS: All subjects demonstrated an increase in contrast thresholds under glare conditions regardless of lens tint. However, brown and yellow lens tints resulted in the least amount of contrast threshold increase. Gray lens tint resulted in the largest contrast threshold increase. CONCLUSIONS: Individuals with lenticular changes may benefit from brown or yellow spectacle lenses under glare conditions. PMID- 12769152 TI - Changes of corneal thickness in patients wearing frequent-replacement contact lenses. AB - PURPOSE: To evaluate changes in corneal thickness after 2, 4, and 6 weeks of frequent-replacement contact lens wearing. MATERIAL AND METHODS: Twenty-eight patients were fitted with ACUVUE (Johnson & Johnson) contact lenses. Central and peripheral corneal thickness was measured before contact lens fitting and after 2, 4, and 6 weeks of wearing. RESULTS: Analysis of variance shows that the period of contact lens wearing has no influence on central corneal thickness. After a small increase in the second week, central corneal thickness returns to the initial values in the fourth and sixth weeks of contact lens wearing. Peripheral corneal thickness was statistically related to the time of contact lens wearing (P < 0.01). A statistically significant difference was observed between the baseline and peripheral corneal thickness measurements in the second, fourth, and sixth week of contact lens wearing (P < 0.01). CONCLUSIONS: Frequent-replacement contact lenses (ACUVUE) cause a small amount of corneal edema, which is higher in the periphery than in the center. Central corneal swelling disappears after 6 weeks of wearing, but in the periphery it remains stable throughout the period of observation. PMID- 12769151 TI - An atypical presentation of Acanthamoeba keratitis in a noncontact lens wearer. AB - This article presents the case of a 49-year-old man who did not have a history of wearing contact lenses and who developed a rapidly progressive course of Acanthamoeba keratitis. The patient developed stromal keratitis that did not respond to herpes simplex virus therapies. Within 1 week after presentation, the patient progressed from mild anterior stromal haze and edema to a ring infiltrate, epithelial loss, and significant corneal edema. Corneal scrapings demonstrated cysts consistent with Acanthanmoeba keratitis. The patient was admitted to the hospital and placed on intensive medical therapy. He responded to therapy, and at 5 months showed central scarring in a quiet eye. This article presents a case of Acanthamoeba keratitis in a non-contact lens wearer, who was diagnosed clinically and histopathologically within 1 week of onset of symptoms. His case was atypical given his lack of contact lens wear or antecedent trauma and rapid progression to a ring infiltrate, usually seen as late findings. PMID- 12769153 TI - Effects of contact lens care solutions on surface exfoliation and bacterial binding to corneal epithelial cells. AB - PURPOSE: The purpose of this study is to assess the effects of commercially available contact lens wetting solutions on bacterial binding and cell exfoliation rates in human corneal epithelium. METHODS: The effects of four contact lens care solutions were tested: ReNu Multi Plus (Bausch & Lomb, Rochester, NY) multipurpose solution; OPTI-FREE Express (Alcon, Ft. Worth, TX) multipurpose solution; Complete Blink-N-Clean (Allergan, Irvine, CA) lens drops; and Lens Plus (Allergan) rewetting drops. STUDY DESIGN: Prospective, double masked, randomized crossover clinical trial (N = 20 subjects). OUTCOME MEASURES: Measures of outcome included binding of Pseudomonas aeruginosa (PA) to exfoliated corneal epithelial cells, and the rate of surface cell exfoliation. Cells were collected at the baseline (pretreatment) examination and 4 days later, after subjects used the assigned solution six times daily and once again immediately before cell collection (posttreatment). Following cell collection, patients underwent 1 week of recovery, during which no drops were used, and random cross over assignment to the next test solution. RESULTS: Use of test solutions increased PA binding, with a range of + 11.9% to + 58.2%. Analyzed together, PA binding increased significantly (+ 29%; P = 0.02, paired t-test); Lens Plus solution alone raised PA binding levels significantly (P = 0.022, 2-way ANOVA, Student-Newman-Keuls [SNK] test). Exfoliation rates were decreased from -7% to 52.7%. Analyzed together, cell exfoliation decreased significantly (P = 0.004; Wilcoxon signed rank test). Individual use of OPTI-FREE decreased exfoliation significantly (P = 0.019: 2-way ANOVA, SNK test). CONCLUSIONS: Topical application of common commercial contact lens care solutions increases PA binding and reduces corneal surface cell exfoliation. Similar effects have also been reported with contact lens wear. Taken together, the data suggest that the use of lens solution itself may play a role in increasing PA binding to corneal epithelial cells and, hence, might potentially contribute inadvertently to increased risk for lens-related microbial keratitis. PMID- 12769154 TI - Photorefractive keratectomy versus laser in situ keratomileusis for the treatment of spherical hyperopia. AB - OBJECTIVE: To compare photorefractive keratectomy (PRK) and laser in situ keratomileusis (LASIK) in treating spherical hyperopia using the VISX STAR S2 excimer laser. INTERVENTION: Fifteen consecutive eyes of 15 patients underwent PRK, and 16 consecutive eyes of 16 patients underwent LASIK (follow-up: 12 months). MAIN OUTCOME MEASURES: Postoperative pain, uncorrected visual acuity (UCVA), deviation from intended correction, and loss of best spectacle-corrected visual acuity (BSCVA). RESULTS: Mean preoperative spherical equivalent was + 2.18 diopter [D] for PRK and + 2.03D for LASIK. All PRK patients experienced significant postoperative pain that required systemic medication, whereas LASIK patients had minor and transient discomfort. Mean deviation from intended correction was -0.83D, + 0.01D, and + 0.18D at 1, 6, and 12 months after PRK, and + 0.22D, +0.30D, and + 0.40D at 1, 6, and 12 months after LASIK (P = 0.002 at 1 month). A higher proportion of LASIK eyes had a UCVA of 20/20 or better at all time points (P = 0.013 and 0.025 at 1 and 3 months, respectively). There was no statistically significant difference between both groups in BSCVA loss. CONCLUSIONS: LASIK and PRK are comparable in efficacy and safety. However, PRK was more painful, with an initial and temporary myopic over-correction that did not occur after LASIK. Stability was achieved between 3 and 6 months following PRK, and one month following LASIK. PMID- 12769155 TI - Preservative-free ocular hydrating agents in symptomatic contact lens wearers: saline versus PVP solution. AB - PURPOSE: To compare two preservative-free hydrating agents, in multidose (ABAK) bottles, in contact lens wearers experiencing symptoms of ocular dryness. METHODS: The endpoint of this 4-week multicenter, randomized, double-blind, parallel-group study comparing a 2% polyvinylpyrrolidone (PVP) solution with a 0.9% NaCl solution was to assess ocular discomfort using a visual analog scale. A biomicroscopic examination and a test of tolerability on instillation were also performed, and adverse events were recorded. RESULTS: Thirty-nine subjects were enrolled (23 PVP; 16 NaCl). The average age was 30; subjects were predominantly female, and mostly wore frequent-replacement contact lenses. They were all exposed to environmental factors such as routine use of video monitors or air conditioning. The two groups were similar at baseline (D0). Both PVP and NaCl improved the comfort of contact lens wear (P = 0.0003), with no difference between them (P = 0.25). The mean daily duration of lens wear and the daily number of instillations to relieve discomfort (4.2 +/- 2.0 for PVP ABAK; 4.6 +/- 1.9 for NaCl ABAK) were comparable. However, PVP use led to more favorable evolution of fluorescein-staining corneal punctuations (P = 0.028). Safety was good, with minimal adverse events considered unrelated to the products. Lens wettability was excellent, and there were no clinically relevant deposits. Most subjects also found the ABAK bottles convenient. CONCLUSIONS: Ocular hydration improves comfort in contact lens wearers. NaCl is an appropriate first-line treatment, but for subjects with fluorescein-staining punctuations, lubrication with PVP is preferable. PMID- 12769156 TI - RK4 (reverse-geometry) contact lens fitting after penetrating keratoplasty. AB - PURPOSE: We evaluated the performance of the RK4 (reverse-geometry) contact lens in patients with flat central and steep peripheral corneal topography after penetrating keratoplasty (PK). METHODS: A retrospective study was performed of post-PK patients fitted with RK4 lenses on the Cornea Service at Wills Eye Hospital, between March 2000 and February 2002. RESULTS: Fifteen eyes of 13 post PK patients were fitted with RK4 lenses and monitored for more than two months (range, 2.1-23.5 months; mean, 8.1 +/- 7.0 months). The average number of lens refits per eye was 1.0 +/- 1.2 (range, 0-4). Ninety-three percent (14/15) of the eyes that were fitted achieved at least 20/30 visual acuity, and 53% (8/15) of eyes achieved 20/20 visual acuity. A comparison of prefitted and post-fitted corrected visual acuity demonstrated a statistically significant improvement (P < 0.01). RK4 lenses were successfully worn full time or part time in all cases. CONCLUSIONS: The RK4 lens can provide well-tolerated visual correction in cases with central flat, peripheral steep corneal contour following PK. PMID- 12769157 TI - Do daily wear opaquely tinted hydrogel soft contact lenses affect contrast sensitivity function at one meter? AB - PURPOSE: To investigate the effects of daily wear opaquely tinted hydrogel soft contact lenses on contrast sensitivity functions, as measured using the Pelli Robson chart viewed at 1 meter. METHODS: Contrast sensitivity was measured in 48 healthy individuals before and 6 hours after soft contact lens wear. Twenty-four subjects used clear soft contact lenses and 24 used tinted soft contact lenses. RESULTS: The contrast sensitivity scores were significantly decreased monocularly and binocularly (P = 0.000 and P = 0.002, respectively) in the colored contact lens group whereas there were no significant changes in the clear contact lens group (P = 0.317 and P = 0.317, respectively). CONCLUSIONS: Color-tinted contact lenses were associated with a reduction of contrast sensitivity function. Therefore, those who wear colored contact lenses should be informed about the possible consequences of tinted contact lens wearing. PMID- 12769158 TI - Clinical results comparing high-oxygen and low-oxygen permeable soft contact lenses in France. AB - PURPOSE: This article reports clinical results comparing a high-oxygen permeable (HDk) silicone hydrogel soft contact lens with a low-oxygen permeable (LDk) soft hydroxyethyl methacrylate contact lens when worn on an extended-wear basis. METHODS: Six practitioners in France enrolled 134 subjects in this 4-month, open label, multicenter, prospective, randomized, cross-over study. Subjects were randomized to receive either an HDk lotrafilcon A or an LDk etafilcon A soft contact lens. Subjects crossed over to the alternative lens after either 3 months' experience with the HDk lens or 1 month's experience with the LDk lens. Clinical and subjective data were collected at scheduled follow-upvisits. RESULTS: The subjective data has been reported in a separate article. Both types of lenses gave similar performance for visual acuity. Biomicroscopic examination findings averaged below grade 1 (trace) for both lenses throughout the study. A statistically significant difference for limbal redness was found at 1 week (HDk = 0.11; LDk = 0.33; paired-t test, P < 0.05). Eight of 10 biomicroscopic signs improved within 1 week when subjects changed from the LDk lens to the HDk lens. Deposits for both lenses remained below grade 1, and wetting was nearly perfect for both lenses throughout the study. Over 90% of fittings for both lenses were acceptable or optimal. CONCLUSION: The improvements in biomicroscopy signs seen with the HDk lens may provide practitioners an alternative to LDk contact lenses for extended wear. PMID- 12769159 TI - Subjective experience with high-oxygen and low-oxygen permeable soft contact lenses in France. AB - PURPOSE: This article reports the subjective results of a study comparing extended-wear high-oxygen permeable (HDk) silicone hydrogel soft contact lenses with low-oxygen permeable (LDk) hydroxyethyl methacrylate lenses. METHODS: Six practitioners in France enrolled 134 subjects in this 4-month, open-label, multicenter, prospective, randomized, cross-over study. Subjects were randomized to receive prescriptions of either an HDk lotrafilcon A or an LDk etafilcon A soft contact lens. Subjects then crossed over to the alternative lens after either 3 months' experience with the HDk lens or 1 month's experience with the LDk lens. Clinical and subjective data were collected at scheduled follow-up visits. RESULTS: The clinical data has been reported in a separate article. After 1 month of wear. HDk lenses were rated better than LDk lenses for all aspects of comfort and overall average symptoms. Subjects were satisfied with both lenses, and lens features were comparable through 1 month. The preference was higher for HDk lenses after crossing over, either from or to LDk lenses. Eighty-six percent of HDk and 85% of LDk subjects reported wearing their lenses while sleeping for 7 nights per week during the study. Seventy-seven percent of HDk and 21% of LDk subjects reported wearing their lenses while sleeping for 22 to 31 nights during the study. CONCLUSIONS: Based on subjective results, therefore, the HDk lens offers another option to patients and practitioners for extended-wear or continuous-wear soft contact lenses. PMID- 12769160 TI - The need for tighter control of cardiovascular risk factors in diabetic patients. AB - Diabetic patients have a two- to four-fold increase in macrovascular disease compared with non-diabetic subjects, with coronary heart disease (CHD) and stroke being the most common causes of death in type 2 diabetes. Diabetic nephropathy has become the most common single cause of end-stage renal disease in industrialized countries. Risk factors, including hyperglycaemia, high blood lipids and high blood pressure (BP), often co-exist in diabetic subjects. One recent metaanalysis, including more than 90,000 patients with a 12.4-year follow up, has demonstrated a continuous increase in the relative risks of morbidity and mortality with increasing blood glucose concentration. Both the Multiple Risk Factor Intervention Trial (MRFIT) and the United Kingdom Prospective Diabetes Study (UKPDS) have confirmed in diabetes the close relationship between total cholesterol levels and elevated risk of cardiovascular events. For every 1 mmol/l increase in low-density lipoprotein cholesterol in type 2 diabetes, the relative risk of CHD increases by 1.57. Furthermore, about 40% of newly diagnosed diabetic patients are also hypertensive. Elevated BP is related to the presence of left ventricular hypertrophy (LVH) and, indeed, LVH is observed in more than 70% of diabetic patients with hypertension. Several studies in diabetes have proven treatment benefits when different risk factors are addressed. The need for tighter control of cardiovascular risk factors in diabetic patients is clear. This may include better control of raised BP, hyperlipidaemia and hyperglycaemia as well as closer monitoring for the appearance of LVH and microalbuminuria. There is a clear need to translate the results of clinical trials into everyday clinical practice. PMID- 12769161 TI - Microalbuminuria: a common, independent cardiovascular risk factor, especially but not exclusively in type 2 diabetes. AB - Microalbuminuria (defined as an albumin-creatinine ratio of 10-25 mg/mmol on the first-morning urine sample, or an albumin excretion rate of 20-200 microg/min on a timed collection) is present in 20-30% of all patients with type 2 diabetes, and is especially common in those with hypertension, endothelial dysfunction and other features of insulin resistance. Although microalbuminuria is predictive of worsening microvascular disease in the kidney (5-10% per year progress to overt diabetic nephropathy), an increased albumin excretion rate (AER) reflects a generalized abnormality of vascular function and is associated with 2-4-fold increases in cardiovascular and all-cause mortality. The extent to which microalbuminuria is a risk factor independent of other variables in type 2 diabetes, e.g. blood pressure and smoking, has been highlighted by recent cohort studies, e.g. the Heart Outcome Prevention Evaluation study and the Wisconsin Epidemiological Study of Diabetic Retinopathy. In the former study, for example, microalbuminuria at baseline increased the adjusted relative risks (RR) of a major cardiovascular event (RR 1.83), all-cause death (RR 2.09) and hospitalization for heart failure (RR 3.23) in both diabetic and non-diabetic subjects. These studies also highlighted that AER is a continuous risk factor, and that levels of AER below the arbitrary threshold for defining microalbuminuria are associated with relatively increased cardiovascular risk. Similarly, microalbuminuria affects 10-15% of middle-aged non-diabetics and is associated with coronary, peripheral and cerebral vascular complications. Detection of microalbuminuria, especially in type 2 diabetes, signifies the need to intensify blood pressure control as part of a multiple risk factor intervention strategy in a high-risk group. As hypertensive patients with type 2 diabetes are frequently treated by more than one antihypertensive agent, ACE inhibitors and low-dose diuretics are preferably recommended in order to provide sufficient blood pressure control and target organ protection. PMID- 12769162 TI - Clinical role of Natrilix SR in the treatment of at-risk hypertensive patients. AB - Although recent trials have shown that antihypertensive treatment can bring about a reduction in stroke, coronary heart disease, heart failure and renal disease, the situation is no longer improving. This is due to the fact that the percentage of hypertensive patients with satisfactory blood pressure is still very poor. International guidelines on hypertension indicate the importance of assessing the absolute risk of patients and the use of a lower dose of drugs to improve the efficacy-tolerability profile. Diuretics used at lower dosage than in the past are effective in reducing morbidity and mortality and continue to be drugs of first choice in the treatment of hypertension. Indapamide sustained release (Natrilix SR) 1.5 mg has an antihypertensive effect equivalent to indapamide immediate release 2.5 mg with a 50% reduction in incidence of serum potassium levels <3.4 mmol/l. Natrilix SR has proved to have a neutral effect both on lipid and glucose profiles and to reduce microalbuminuria in diabetic hypertensive patients. Recent multicentre European clinical trials have shown that Natrilix SR decreases diastolic blood pressure to <90 mmHg in about 75% of patients treated for 1 year. In elderly patients with isolated systolic hypertension, Natrilix SR has been proven to be as effective as amlodipine 5 mg and significantly more effective than hydrochlorothiazide 25 mg. Natrilix SR produces regression of left ventricular hypertrophy which, in the Left ventricular hypertrophy: Indapamide Versus Enalapril study was greater than that induced by enalapril. Natrilix SR represents an appropriate choice not only as a first-line drug in many hypertensive patients but also in at-risk patients like the elderly, subjects with other cardiovascular risk factors, target organ damage, diabetes, or impaired renal function. PMID- 12769163 TI - Effect of indapamide SR on microalbuminuria--the NESTOR study (Natrilix SR versus Enalapril Study in Type 2 diabetic hypertensives with micrOalbuminuRia)- rationale and protocol for the main trial. AB - In type 2 diabetic hypertensive patients, microalbuminuria can be due to hypertension and/or diabetic nephropathy. Angiotensin-converting enzyme (ACE) inhibitors act preferentially on microalbuminuria due to diabetic nephropathy. The objective is to demonstrate the efficacy of a thiazide-like diuretic, indapamide sustained release (SR), at reducing microalbuminuria in hypertensive type 2 diabetic patients in comparison with an ACE inhibitor, enalapril. The study is an international multicentre, 12-month, randomized, double-blind, controlled, two parallel group study of type 2 diabetic patients with hypertension (140 mmHg < or = systolic blood pressure <180 mmHg and diastolic blood pressure <110 mmHg) and microalbuminuria. Intervention is after a 4-week placebo period, patients with microalbuminuria > or = 20 and < or = 200 microg/min are randomized to indapamide SR 1.5 mg or to enalapril 10 mg once a day for a one-year treatment period. An additional label treatment by amlodipine 5-10 mg (1st step) and atenolol 50-100 mg (2nd step) a day is permitted after 6 weeks of treatment based upon blood pressure response. The main outcome measures are microalbuminuria expressed as urinary albumin to creatinine ratio, albumin fractional clearance, and albumin excretion rate evaluated on overnight urine collections. Secondary criteria are supine and standing systolic, diastolic and mean blood pressure; and biological and clinical safety. This study will complete the knowledge of the efficacy of indapamide SR in hypertension and target organ damage and will provide valuable information on the management of type 2 diabetic hypertensives with microalbuminuria. PMID- 12769164 TI - New treatment guidelines for a patient with diabetes and hypertension. AB - Guidelines for medical treatment are becoming increasingly popular and many guidelines have been produced by various societies in diabetes, hypertension, and renal disease as well as general medicine. By their nature, they are outdated considering the rapid and efficient publication of many papers related to the treatment of hypertension in diabetes. Increased blood glucose causes vascular damage and abnormal vascular structure all over the body, an abnormal structure that is especially vulnerable to high blood pressure, even within the so-called normal range. There is now more and more evidence, especially in diabetics, that blood pressure should be as low as possible. In this context, it is important to stress that the so-called J-shaped relationship between blood pressure and mortality may not be so relevant. Major epidemiological studies came from the Framingham and the Multiple Risk Factor Intervention Trial (MRFIT) Diabetic Cohort. The MRFIT Cohort showed that cardiovascular mortality was increased by a factor of 2-4 in diabetic patients, and there was a clear association between systolic blood pressure and complications without any threshold value. It could be suggested that since diabetes is an important cardiovascular risk factor, a lower value (130/85 mmHg) than for non-diabetics (140/90 mmHg) should be proposed. The tight blood pressure control arm of the United Kingdom Prospective Diabetes Study was <150/85 mmHg (achieved 144/82 mmHg) and the aim in the less tight control arm was <180/105 mmHg (achieved 154/87 mmHg). In the tight control group, 29% needed three or more antihypertensive drugs. In the Hypertension Optimal Treatment study, the frequency of major cardiovascular disease events in the group with target <80 mmHg (achieved 144/81 mmHg) was 11.9/1000 patients/year, which was significantly lower than the event rate (24.4/1000 patients/year) in the group with target <90 mmHg (achieved 148/85 mmHg). A reduction in the frequency of diabetic nephropathy by angiotensin-converting enzyme (ACE) inhibitor treatment in normotensive lean microalbuminuric type 2 diabetic patients has been shown. However, it is impossible from the present data to draw any conclusions with respect to effect on the main composite endpoint of ACE inhibition in microalbuminuric type 2 diabetic patients without previous cardiovascular events or without hypertension. Recent published studies have also demonstrated beneficial effects with angiotensin receptor blockers (ARBs) in hypertensive patients with type 2 diabetes and nephropathy. Diuretics form a very important basis for antihypertensive treatment, also often in combination with agents that inhibit the renin-angiotensin system. Several studies show that treatment with the diuretic indapamide reduces the level of microalbuminuria in patients with type 2 diabetes. Diuretics were used as an adjunctive to reduce blood pressure in all studies; it is therefore understandable that many guidelines suggest that diuretics form part of the treatment of hypertension in diabetics. Many studies of an epidemiological nature and follow-up studies in diabetic patients show that blood pressure control is of vital concern in the prevention of diabetic complications, and indeed the usual criteria for good blood pressure control may not be stringent enough in diabetic patients. Many classes of antihypertensives may be used, but it appears that diuretics, such as indapamide sustained release (SR), constitute an important proposal in all treatment strategies. PMID- 12769165 TI - The role of functional capacity evaluations in occupational health settings. PMID- 12769166 TI - The North Country on the Job Network: a unique role for occupational health nurses in a community coalition. AB - 1. Through a community based program, nurses were hired by a rural medical center to facilitate care for injured workers in the community. This placed the nurses outside the industrial and insurance arenas. 2. Rapid access to care and expedited return to work were made possible largely through improved communication, facilitated by the nurse case coordinators, among all involved parties (i.e., workers, employers, health care providers, insurance carriers). 3. The program provides access to occupational health nurses, or case managers, to a large number of rural workers--many of whom work for small employers and would not have these services otherwise. 4. The simple administrative model has resulted in increased numbers of workers returning to work with decreasing numbers of transitional or modified duty days. PMID- 12769167 TI - Lifting teams in health care facilities: a literature review. AB - 1. Manual lifting and transfer activities are job tasks frequently associated with back injuries in nursing personnel. One approach with potential to decrease these injuries is the lifting team. 2. In program evaluations completed to date, there have been numerous benefits and several limitations attributed to use of lifting teams in health care facilities. 3. Benefits of lifting teams include reductions in lost time workdays, restricted workdays, workers' compensation claims, and injuries to lifting team members; satisfaction of patients, staff, and lifting team members; and capacity of the lifting team to absorb the majority of high risk lifts and transfers on shifts in which they operate. 4. Lifting teams may not be appropriate for all settings, require infrastructure and lifting team equipment to support their use, and require careful consideration related to staffing. However, when their use is appropriate, efforts to overcome their limitations can be accomplished with careful evaluation of outcome measures and indicators. PMID- 12769168 TI - Cardiovascular disease risk factors and the perception of general health among male law enforcement officers: encouraging behavioral change. AB - The relationship among cardiovascular disease (CVD) morbidity, risk factors (including stress), and the perception of health among male law enforcement officers (LEOs) compared to men in the general population were examined in this study. Self reported prevalence of CVD and CVD risk factors among currently employed male LEOs from nine states (n = 2,818) were compared to those of other men in the same states (n = 9,650 for CVD risk factors, n = 3,147 for CVD prevalence). Perceived stress in LEOs was assessed to determine if it affected the relationship between CVD prevalence and CVD risk factors. Cross tabulated simple percentages showed CVD was less prevalent in the LEO group than among the general population. The best predictor variables for CVD were perceived stress, time in the profession, and hypertension. The LEO group had greater prevalence of hypercholesterolemia, overweight, and tobacco use than the general population. However, a greater percentage of LEOs perceived their health as "good to excellent" compared to men in the general population. Using multivariate analysis of variance (MANOVA) it was determined that perceived stress was associated with CVD in the LEO group and three CVD risk factors (i.e., cholesterol, hypertension, physical activity) were significantly affected by perceived stress. Among susceptible officers, stress may contribute to CVD development as well as potentiate several CVD risk factors. However, an apparent lack of association exists between perception of general health and CVD risk in LEOs. PMID- 12769169 TI - Preparing for smallpox: occupational health nursing update. AB - Smallpox has plagued humans for thousands of years. Although smallpox has been eliminated as an active disease in humans, there is a threat that terrorist organizations or nations could use smallpox as a weapon against the population of the United States. The U.S. government has recognized this threat and has asked the CDC to prepare for such a possibility. A plan to identify, treat, and vaccinate the population in any area of the country where an outbreak of smallpox might occur has been developed and is being implemented. The CDC plan calls for rapid response by local, state, and federal response teams. Preparation of these teams is currently underway. Parts of the CDC plan, including educating and screening individuals who may be part of the response teams, vaccinating personnel, and following up with the vaccinated personnel, are activities consistent with the occupational health nurse's role. PMID- 12769170 TI - Working with fibromyalgia. PMID- 12769171 TI - The neuronal structure of the red nucleus in newborn guinea pigs. AB - The preparations, stained according to the Nissl and Kluver-Barrera methods, were used to describe the topography and morphology of the red nucleus (RN) as well as the structure of the rubral perikarya in newborn (P0) guinea pigs. The Golgi impregnated preparations were used to distinguish types of neurons. RN is a uniform cell group and has the length from 740 to 860 microm. The Nissl stained perikarya were classified into three categories: big, medium-sized and small (A, B, C, respectively). The big perikarya, which contain a lot of tigroidal substance, were mainly observed at the caudal and ventral portions of RN. The small perikarya often have multiple nucleoli. The impregnated neurons were classified into 5 types: 1--large, aspiny, rich-arborised multipolar cells, 2- large and medium sized, spiny, rich-arborised fusiform or pear-shaped cells, 3- medium-sized, spiny, rich-arborised rounded cells, 4--medium-sized, spiny, rich arborised bipolar cells, 5--small and single medium-sized cells. The 5th type constitutes a heterogeneous population and also has neurons in different developmental stages. Intraspecies variations concerning both the length of RN and a number of the triangular perikarya of the red nucleus were observed in the examined guinea pigs. PMID- 12769172 TI - Postnatal development of NOS-ir neurons in the rat claustrum. AB - The morphological features of nitric oxide synthase (NOS)-containing neurons in the rat claustrum (Cl) were studied during the period of four months after birth. Forty-five animals divided into nine groups, according to survival period (P0, P4, P7, P10, P14, P21, P28, P60, P120) were used in the study. The immunocytochemical staining to neuronal NOS was performed and the material was studied both qualitatively and quantitatively using unbiased stereological methods. Our observations indicate that the process of maturation of NOS immunoreactive (ir) neurons in Cl takes place during the early postnatal period. We report the increase of numerical density of immunoreactive neurons, changes in neuronal size, expressed by the decrease of the percentage of small neurons with simultaneous increase of the participation of medium-sized neurons and large neurons. In the whole studied period the prevalence of oval and fusiform neurons is observed. However, the increase of the proportion of multipolar neurons takes place. Round neurons are most characteristic in the youngest groups of animals and later become dominated by the developing subpopulations of ir neurons of other shapes. In the anterior, central and posterior parts of Cl, a similar pattern of maturation of NOS-ir neurons is observed. No prevalence of characteristically shaped neurons is observed in any part of Cl. The adult-like pattern of morphological features in the NOS-ir neuronal population in Cl is reached in the third postnatal week. The maturation of NOS-ir neurons in the claustrum is a dynamic process which is not stabilised at the moment of birth. It may be assumed that characteristic changes of the NOS-ir population of neurons may be influential on the physiological processes observed in Cl. These may in particular have some importance for the processes of synaptogenesis and establishing as well as refining of numerous claustral connections with the other structures of the central nervous system. PMID- 12769173 TI - Immunohistochemical and morphometric study of the effect of maternal diabetes on rat foetal pancreatic islets. AB - The effect of maternal diabetes on the foetal endocrine pancreas has been the subject of extensive studies, but a detailed quantitative immunohistochemical investigation is not available. This study was therefore undertaken to investigate the effect of gestational diabetes on the morphology of foetal rat islets. Sections were stained with anti-insulin (B cells) antibodies and were used for morphometric analysis. The B cell volume density was significantly lower in the diabetic group, while islet volume density, islet diameter, islet volume and absolute islet cell numbers were significantly greater in the diabetic group. The B cell nuclear diameter and volume were not significantly different in the diabetic group. The results obtained from this investigation indicate that maternal diabetes induces foetal islet hypertrophy and causes an increase in the total islet cell number. PMID- 12769174 TI - An evaluation of the reproducibility of the measurement of the intima-media thickness of carotid arteries. AB - The intima-media thickness (IMT) of carotid arteries was demonstrated to be a reliable measure for early stages of atherosclerosis. B-mode ultrasound may be used to measure carotid IMT. The measurements of the IMT of the carotid artery (CA) conducted by different investigators can be comparable and enable the implementation of clinical trial successfully while maintaining a high reproducibility value. The objective of the study was to evaluate the reproducibility of the measurements made by the same investigator on two separate occasions (intraobserver variability) and the reproducibility of the off-line measurements between four sonographers in our laboratory (interobserver variability). The IMT of CA in 25 subjects (15 post stroke and 10 healthy persons) was investigated with the use of high-resolution ultrasonography. The CA subdivided into the common, bulbs and internal segments were scanned twice with a 3-week interval. Additionally three other readers with different levels of experience and skills in ultrasonography were asked to perform the same measurements in duplicate with at least a 3-week interval between. A high concurrence for intraobserver variability was detected with a correlation coefficient ranging from 0.92 to 0.95; p < 0.0001, and maximal bias 0.019 mm. Interobserver variability for all four readers also demonstrated a high correlation coefficient ranging from 0.72 to 0.83; p < 0.0001, and the maximal bias of measurements did not exceed 0.08 mm. The analogue measurements performed by the team demonstrate a reliable reproducibility in terms of the results of morphologic measurements. The differences obtained in the study were less than the error of the method (i.e. 0.1 mm) and should not influence clinical decision making. Additionally, this study demonstrated that interobserver concurrence increases with the increasing experience of the investigators. PMID- 12769175 TI - Sex dimorphism in development dynamics and in development progression of morphological features in human foetuses. AB - The study material comprised 3889 foetuses of both sexes, aged 20-42 weeks. t Student test has been applied to evaluate the existence of potential sex dependent differentiation of developmental trends as assessed by weekly measurements of selected somatic features and by the weight of internal organs. The regression coefficients of the analysed variables have been compared against the opposite sex. The rate of development of the analysed features in consecutive weeks has been found to be sex-related. Highest sex-related differences have been observed for the total body weight and for the weight of internal organs, with the exception of the adrenals, and the differences have been significant enough to justify the existence of contrasting, sex-dependent patterns of development of the analysed variables. The development of the analysed morphological features has been depicted by curvilinear regression. When described by various degree polynomials the development course of the analysed features displays sex-related differences. Only the change in the weight of the adrenals is similar for both sexes. The evaluation of the developmental advancement of the analysed features has revealed that they are usually at a more advanced development level in female foetuses. PMID- 12769176 TI - Developmental interdependence between selected somatic features and the weight of internal organs in human male and female foetuses. AB - The study material comprised 3889 foetuses of both sexes, aged 20-42 weeks and focused on developmental interdependence between the weight of the internal organs and their relation to selected somatic features during the prenatal period. The study also attempted to distinguish potential sex-related differences in the degree of statistical interdependence between the analysed variables. As a result, the analysis of linear and canonical correlation coefficients for consecutive weeks of foetal life has been carried out. High correlation coefficients have been obtained which indicate strong developmental interdependence between those organs and the somatic features. However, no significant sex-related differences in the developmental interdependence of the analysed variables have been observed. PMID- 12769177 TI - Correlation of heart external dimensions with body external dimensions of human foetuses. AB - In an evaluation of foetal hearts in II and III pregnancy semester, correlation with exterior foetal dimensions should be taken into consideration. It is interesting also if all exterior heart dimensions are correlated at the same level with overall body dimensions. The subject of examination was 204 foetuses of both sexes, including 106 males and 98 females, in 5th and 6th month of foetal life. Foetuses were fixed for minimum 3 months in 9% formalin solution. They were taken from natural abortions, without exterior features of developmental malformations. Examined material was evaluated in month groups of morphological age. The number of foetuses in different groups was various. Exterior heart dimensions were considered: height, width, heart depth, and also heart circumference in coronary sulcus (atrial-ventricular). Exterior dimensions of foetuses were: vertex-plantare, vertex-tubulare. All exterior dimensions of heart are closely and strongly correlated with both exterior dimensions of foetuses. PMID- 12769178 TI - Histomorphometric study of megakaryocytes in bone marrow in selected myeloproliferative and lymphoproliferative diseases. AB - The aim of the study was an estimation of the histomorphometry of megakaryocytes (MK) in bone marrow in selected myeloproliferative and lymphoproliferative diseases. Bone marrow specimens were obtained by trephine biopsies from 41 patients with polycythaemia rubra vera (PV), idiopathic myelofibrosis (MF), chronic lymphocytic leukaemia (CLL), hairy cell leukaemia (HCL) and diffuse large B-cell lymphoma (L). Morphometric evaluation was performed using a standard program set MicroImage (OLYMPUS). The greatest number of typical nucleated MK, "naked" nuclei, anucleated cytoplasmic fragments and the largest area were found in PV. The circular deviation factor of MK and their nuclei increased in all cases. The greatest number of clusters was observed in PV and HCL. A significant increase in the number of dysplastic and "naked" nuclei of MK was noted in all selected haematological diseases. The presence of neoplastic cells in bone marrow increased the morphological changes in MK. Quantitative and morphometrical significant differentiation of MK in separate microscopic field in the same slides confirms the necessity of performing trephine biopsies in each patient with haematological disorders. PMID- 12769179 TI - The examinations of thyroid gland isthmus topography in foetal period development --practical importance. AB - The studies aimed at following isthmus of thyroid position changes in particular months of its intrauterine development. Survey material consisted of 124 human foetus thyroids aged 4-7 months of intrauterine development. The basic examination method was classical anatomical preparation. Three aspects were considered in thyroid isthmus position assessment: holotopy, skeletotopy and syntopy. The gland holotopy analysis was carried out with the usage of isthmus position indices elaborated especially for the studies. The results were statistically analysed. Thyroid isthmus was observed to change its position. Although statistically significant differences in thyroid isthmus position were not proved, some tendencies of the one-position changes were detected, especially of larynx and trachea skeletons. The tendencies were based on a relatively more significant lowering of isthmus in comparison with gland lateral lobes. The basic reason for thyroid isthmus position changes in the development period is a disproportion between foetus gland and cervix growth rates. PMID- 12769180 TI - Dimension of circumflex branch according to type of heart vascularisation in foetal life of human. AB - Typological differentiation of coronary arteries binds to various areas of vascularisation, which could suggest essential differences between vasculatory dimensions. There are not many papers analysing the influence of heart vascularisation on epicardial vessel dimensions during postnatal human life, furthermore, there are hardly any papers dealing with the prenatal period. The subject of examination was 188 human foetuses from 4th to 7th month of prenatal life. Foetuses were fixed for minimum 3 months in 9% formalin solution. They were taken from natural abortions and did not characterise any external malformations. There was a different number of foetuses in a variety of morphological age groups. Adachi classification was used to describe all types: type I--classical, with equal coronary arteries, type II--predominance of right coronary artery, type III--predominance of left coronary artery. Speed of circumflex branch growth in different types of vascularisation is various. Analysis of the differences among dimensions of artery in various types showed there are statistically crucial ones, especially between: types III and I or types III and II. PMID- 12769181 TI - The topography of the subthebesian fossa in relation to neighbouring structures within the right atrium. AB - The majority of anatomical structures within the heart during typical atrial flutters' ablation, right sided accessory pathway ablation or slow pathway ablation are invisible or blurred. Therefore it is very important to know in details interior right atrial structures during such procedures. In the neighborhood of coronary sinus orifice small concavity is visible. This area, called subthebesian fossa, is placed between the os of coronary sinus, the orifice of vena cava inferior and tricuspid annulus. The fossa is on the way of typical atrial flutters' reentrant circuit and is placed next to the isthmus area, which has become a target site for ablative therapy. Regarding the facts mentioned above we decided to examine the topography of this concavity in relation to neighboring structures. Research was conducted on material consisting of 45 human hearts of both sexes, from 19 to 71 years of age. The hearts came from patients whose death was not cardiologic in origin. The topography of the fossa was examined in relation to coronary sinus orifice (diameter A), vena cava inferior orifice (diameter B) and the attachment of the posterior leaflet of the tricuspid valve (diameter C). Besides we measured two perpendicular sizes in the inlet plane of the fossa. There were the longest size (diameter D) and the shortest size of the fossa (diameter E). We also defined deepness of the fossa (diameter F). Diameter A was from to 2 to 7mm (avg. 4.9 +/- 1.4 mm), diameter +/- from 2 to 8mm (avg. 4.0 +/- 1.6 mm) and diameter C from 5 to 9 mm (avg. 7.0 +/- 1.5 mm). The longest size in inlet plane of the concavity (diameter D) was from 12 to 18 mm (avg. 14.1 +/- 1.7 mm) and shortest size (diameter E) was from 7 to 14 mm (avg. 9.0 +/- 1.7 mm). The deepness of the fossa (diameter F) was from 2 to 7 mm (avg. 4.8 +/- 1.2 mm). The subthebesian concavity is inconstant anatomical structure, occurring in all forty five examined hearts (100%). The shape and sizes of the subthebesian fossa were variable in examined group of hearts. Our data suggest that differences in diameters between subthebesian fossa and neighboring structures may have clinical importance during ablation procedure. PMID- 12769182 TI - Neurosurgical stereomorphometry in vivo--method description and error measurement. AB - Rules of geometry and stereomorphometry are often applied to narrow and deep neurosurgical approaches. Methods of research are based on the direct cadaver measurements, radiological analysis and intraoperative measurements. Newly developed devices allow direct morphometry to be performed in vivo, during the operation. We describe the use of the neuronavigation system Stealth Station by Medtronic for such stereomorphometric measurements and evaluate the precision of the described method. PMID- 12769183 TI - The changing pattern of HIV neuropathology in the HAART era. AB - Highly active antiretroviral treatment (HAART), which has been available for most AIDS patients in France since 1996, has resulted in a dramatic improvement of the progression of the disease. From the survey of our series of 343 brains with acquired immunodeficiency syndrome (AIDS) from patients who died between 1985 and 2002, we found both quantitative and qualitative changes in the pattern of human immunodeficiency virus (HIV) neuropathology. Quantitatively, despite a dramatic decrease in the number of autopsies, brain involvement remained a major cause of death. There was an overall decrease in incidence of cerebral toxoplasmosis, cytomegalovirus encephalitis (CMVE), and HIV encephalitis (HIVE), for which successful treatment is available. This contrasted with the unchanged incidence of progressive multifocal leukoencephalopathy (PML) and malignant non-Hodgkin lymphomas (MNHL). However, when looking closer at the 3 last years, the incidence of diseases affecting patients with severe immunodepression (CMVE, PML, and MNHL) decreased between 2000 and 2002, whereas infections occurring in patients with milder immunodeficiency, toxoplasmosis, varicella-zoster encephalitis (VZVE), or herpes simplex virus encephalitis (HSVE) became more frequent. In addition, we found uncommon types of brain infection such as BK virus encephalitis or general paresis. Finally, we described new variants of HIVE: severe leukoencephalopathy with intense perivascular macrophage and lymphocyte infiltration, possibly due to an exaggerated response from a newly reconstituted immune system, and chronic "burnt out" forms of HIVE as VZVE, toxoplasmosis, or PML, possibly associated with prolonged survival, in which neither inflammation nor organisms could be detected. These findings are compared with those reported in other neuropathological studies from different developed countries. PMID- 12769184 TI - Nitrosative and oxidative injury to premyelinating oligodendrocytes in periventricular leukomalacia. AB - Periventricular leukomalacia (PVL), the major substrate of cerebral palsy in survivors of prematurity, is defined as focal periventricular necrosis and diffuse gliosis in immature cerebral white matter. We propose that nitrosative and/or oxidative stress to premyelinating oligodendrocytes complicating cerebral ischemia in the sick premature infant is a key mechanism of injury interfering with maturation of these cells to myelin-producing oligodendrocytes and subsequent myelination. Using immunocytochemical markers in autopsy brain tissue from 17 PVL cases and 28 non-PVL controls, we found in the PVL cases: 1) selective regionalization of white matter injury, including preferential involvement of the deep compared to intragyral white matter; 2) prominent activation of microglia diffusely throughout the white matter; 3) protein nitration and lipid peroxidation in premyelinating oligodendrocytes in the diffuse component; 4) preferential death of premyelinating oligodendrocytes diffusely; and 5) virtual sparing of the overlying cerebral cortex, as demonstrated by markers of activated astrocytes and microglia. These data establish that PVL is primarily a white matter disease that involves injury to premyelinating oligodendrocytes, potentially through activation of microglia and release of reactive oxygen and nitrogen species. Agents that prevent nitrosative and oxidative stress may play a key role in ameliorating PVL in premature infants in the intensive care nursery. PMID- 12769185 TI - Lysosomal activation is a compensatory response against protein accumulation and associated synaptopathogenesis--an approach for slowing Alzheimer disease? AB - Previous reports suggest that age-related lysosomal disturbances contribute to Alzheimer-type accumulations of protein species, blockage of axonal/dendritic transport, and synaptic decline. Here, we tested the hypothesis that lysosomal enzymes are upregulated as a compensatory response to pathogenic protein accumulation. In the hippocampal slice model, tau deposits and amyloidogenic fragments induced by the lysosomal inhibitor chloroquine were accompanied by disrupted microtubule integrity and by corresponding declines in postsynaptic glutamate receptors and the presynaptic marker synaptophysin. In the same slices, cathepsins B, D, and L, beta-glucuronidase, and elastase were upregulated by 70% to 135%. To address whether this selective activation of the lysosomal system represents compensatory signaling, N-Cbz-L-phenylalanyl-L-alanyl diazomethylketone (PADK) was used to enhance the lysosome response, generating 4- to 8-fold increases in lysosomal enzymes. PADK-mediated lysosomal modulation was stable for weeks while synaptic components remained normal. When PADK and chloroquine were co-infused, chloroquine no longer increased cellular tau levels. To assess pre-existing pathology, chloroquine was applied for 6 days after which its removal resulted in continued degeneration. In contrast, enhancing lysosomal activation by replacing chloroquine after 6 days with PADK led to clearance of accumulated protein species and restored microtubule integrity. Transport processes lost during chloroquine exposure were consequently re-established, resulting in marked recovery of synaptic components. These data indicate that compensatory activation of lysosomes follows protein accumulation events, and that lysosomal modulation represents a novel approach for treating Alzheimer disease and other protein deposition diseases. PMID- 12769186 TI - Inflammatory myopathy with abundant macrophages (IMAM): a condition sharing similarities with cytophagic histiocytic panniculitis and distinct from macrophagic myofasciitis. AB - We describe the unreported pattern of inflammatory myopathy with abundant macrophages (IMAM) as a main differential diagnosis of postimmunization aluminum hydroxide-induced macrophagic myofasciitis (MMF). IMAM was mainly detected among patients with a dermatomyositis (DM)-like disease. Among 113 muscle biopsies from DM patients collected from 1974 to 2000, intensity of macrophage infiltration was highly variable: 41.5% (-/+); 34.5% (+); 17% (++): and 7% (+++). The 27 patients from groups (++) and (+++) had a similar pattern of macrophagic infiltration and were considered to have IMAM. They were compared to 40 MMF patients. In IMAM, macrophage infiltrates were diffuse and correlated positively with both T cell infiltrates and acute muscle fiber damage, and showed pictures of hemophagocytosis (21/27). Connective tissue structures were infiltrated by noncohesive, ribbon-forming collections of large basophilic macrophages containing no crystalline inclusions. In MMF, macrophage infiltrates were focal and formed compact well-delineated aggregates of granular PAS+ cells, loaded with crystalline aluminum hydroxide particles, in the absence of either hemophagocytosis or conspicuous muscle damage. Review of the literature indicates similarities between IMAM and "cytophagic histiocytic panniculitis" (CHP), a condition characterized by T cell-triggered macrophage hyperactivation. Both IMAM and CHP, but not MMF, may be associated with a life-threatening hemophagocytic syndrome. PMID- 12769187 TI - Expression of excitatory amino acid transporter-1 in brain macrophages and microglia of HIV-infected patients. A neuroprotective role for activated microglia? AB - Recent experimental studies showed that activated macrophages/microglia (AMM) express excitatory amino acid transporters (EAATs), suggesting that, in addition to their neurotoxic properties, they also have a neuroprotective role by clearing extracellular glutamate and producing antioxidant glutathione. To test this hypothesis in human, the brain of 12 HIV-positive patients and 3 controls were immunostained for EAAT-1. EAAT-1 was expressed by AMM in all HIV-infected cases but not in HIV-negative controls. Expression varied according to the disease stage. In 5 cases with active HIV-encephalitis (HIVE), AMM strongly expressed EAAT-1 in the white matter and basal ganglia, analogous to HLA-DR and CD68 expression. There was weaker expression in the cortex and perineuronal microglial cells were not involved. In a case with "burnt out" HIVE following highly active antiretroviral therapy (HAART), EAAT-1 expression was mild, identical to that of HLA-DR and CD68 in the white matter and cortex and involved perineuronal microglial cells. In 3 AIDS patients without HIVE and in 3 pre-AIDS cases, EAAT-1 expression in the white matter was weaker than HLA-DR and CD68 expression; there was stronger correlation in the gray matter where perineuronal microglial cells were stained predominantly. Our findings in humans tend to confirm that AMM, particularly perineuronal microglial cells, play a neuroprotective role in the early stages of HIV infection and, possibly, following treatment. This is in keeping with the early microglial activation seen in pre-AIDS cases, and the late occurrence of neuronal loss. It may also explain the reversible cognitive disorders following treatment in some cases. PMID- 12769188 TI - Neurotoxicity of acetaldehyde-derived advanced glycation end products for cultured cortical neurons. AB - The Maillard reaction that leads to the formation of advanced glycation end products (AGEs) plays an important role in the pathogenesis of angiopathy in diabetic patients, in aging, and in neurodegenerative processes. We hypothesize that acetaldehyde (AA), one of the main metabolites of alcohol, may be involved in alcohol-induced neurotoxicity in vivo by formation of AA-derived AGEs (AA AGEs) with brain proteins. Incubation of cortical neurons with AA-AGE produced a dose-dependent increase in neuronal cell-death, and the neurotoxicity of AA-AGE was neutralized by the addition of an anti-AA-AGE-specific antibody, but not by anti-N-ethyllysine (NEL) antibody. The AA-AGE epitope was detected in human brain of alcoholism. We propose that the structural epitope AA-AGE is an important toxic moiety for neuronal cells in alcoholism. PMID- 12769189 TI - Aberrant neuronal migration in the brainstem of fukuyama-type congenital muscular dystrophy. AB - We examined the brainstem of 10 patients with Fukuyama-type congenital muscular dystrophy (FCMD). In the midbrain we noted leptomeningeal glioneuronal heterotopia (LGH) (n = 9) and intramural "micropolygyria" (n = 1) in the tectum, as well as tyrosine hydroxylase-positive ectopic neurons/fibers ventral to the cerebral peduncle (n = 3). In the pontomedullary region, glial fibrillary acidic protein-positive subpial tissue intermingled with neurons and myelinated fibers was present in the ventrolateral pontine surface in all cases and extended over the lateral surface of the upper medulla oblongata. This subpial gliotic band was often contiguous with the extra-pial LGH tissues. The gliotic band protruded from the ventrolateral pontine surface in 3 cases and appeared to include ectopic neurons of the pontine nucleus. Disarrangement of the arcuate nuclei (n = 3) was also noted in the medulla oblongata. We hypothesize that both the radial and tangential neuronal migration systems are disrupted in the FCMD brainstem in addition to altered neuronal migration in the cerebral and cerebellar cortex. Fukutin protein may play a part in the morphogenesis of certain neuronal structures in the brainstem and the dysplastic structure termed "aberrant pyramidal tract" in previous reports may essentially result from an ectopic migration of pontine nucleus neurons. PMID- 12769190 TI - Differential lipid peroxidation, Mn superoxide, and bcl-2 expression contribute to the maturation-dependent vulnerability of oligodendrocytes to oxidative stress. AB - To understand the basis of oligodendrocyte (OL) susceptibility to oxidative injury, purified rat OL cultures at different stages of maturation were exposed to nitric oxide (NO) donors with fast or slow kinetics of release and to tert butyl-hydroperoxide, a membrane-permeant organic hydroperoxide. OL precursors (pre-OL) displayed the highest vulnerability to both oxygen or nitrogen reactive species, whereas mature OLs were uniquely vulnerable to long-lasting levels of NO. Cell death occurred by necrosis as well as apoptosis associated with increased caspase-3 activity and, only in the case of pre-OLs, with a decreased expression of the anti-apoptotic protein bcl-2. Pre-OLs were also more susceptible than mature OLs to lipid peroxidation, as measured by F2-isoprostane content in culture media. Finally, pre-OLs, but not mature OLs, expressed high levels of the mitochondrial scavenging enzyme Mn superoxide dismutase, suggesting that pre-OLs may efficiently convert anion superoxide into hydrogen peroxide and, paradoxically, be more predisposed than mature OLs to a toxic imbalance between hydrogen peroxide production and detoxification processes. These data suggest that susceptibility to lipid peroxidation, expression of the scavenging enzyme Mn superoxide dismutase and of the anti-apoptotic protein bcl-2, may contribute to the maturation-dependent vulnerability of OLs to oxidant injury. PMID- 12769191 TI - Lysophosphatidic acid promotes the proliferation of adult Schwann cells isolated from axotomized sciatic nerve. AB - We have previously found that adult Schwann cells express receptors for lysophosphatidic acid (EDG2, EDG7) and sphingosine-1-phosphate (EDG5) and that expression of these receptors is significantly upregulated in injured sciatic nerve coincident with postaxotomy Schwann cell proliferation. Based on these observations, we hypothesized that lysophosphatidic acid and/or sphingosine-1 phosphate promote Schwann cell mitogenesis in injured adult nerve. We found that both saturated and unsaturated forms of lysophosphatidic acid, but not sphingosine-1-phosphate, induce DNA synthesis in adult Schwann cells isolated from surgically transected sciatic nerve. Lysophosphatidic acid induces adult Schwann cell DNA synthesis in a dose-dependent manner, acting at 0.1- to 10 microM concentrations. Lysophosphatidic acid-mediated stimulation of adult Schwann cell DNA synthesis occurs via a signaling pathway involving a pertussis toxin-sensitive (G(i)/G(o)) G-protein. Activation of phosphatidylinositol-3 kinase, cAMP-dependent protein kinase A and mitogen-activated protein kinase kinase is also required for lysophosphatidic acid-induced Schwann cell mitogenesis. These findings demonstrate that lysophosphatidic acid promotes proliferation of adult Schwann cells isolated from injured nerve and are consistent with the hypothesis that lysophosphatidic acid promotes in vivo Schwann cell mitogenesis in regenerating peripheral nerve. PMID- 12769192 TI - Pediatric oligodendrogliomas: a study of molecular alterations on 1p and 19q using fluorescence in situ hybridization. AB - Oligodendrogliomas (OGs) are rare in children and have not been well characterized from a molecular viewpoint. In adults, losses on chromosomes 1p and/or 19q are common in "oligodendroglial" neoplasms and are highly associated with chemosensitivity and greater length of survival, especially in the anaplastic category. We have analyzed the 1p/19q status of pediatric OGs and compared it with similar alterations in adult OGs. Paraffin sections from 26 pediatric OGs (21 WHO Grade II OGs: 2 anaplastic oligodendrogliomas [AOGs]: and 3 mixed oligo-astrocytomas [MOA]) were retrieved. Fluorescence in situ hybridization (FISH) was performed using probes spanning the 1p32 and 19q13 regions. In tumors from children 0 to 9 years of age (n = 15), none had any deletions on 1p or 19q, but 2 had polysomies for 1p and/or 19q. All are alive and 4 have had recurrences. In tumors from children > 9 years, losses were identified on chromosomes 1p (5/11; 45%) and/or 19q (3/11; 27%), but to a much lesser extent than that observed in adult OGs. Tumors from 6 older patients also had polysomies for 1p and/or 19q. Although the majority of the older children are alive, 4 had recurrences. Curiously, 2 of the older children with AOGs had combined losses and polysomies on 1p and 19q, but responded poorly to treatment and died within a year. We conclude that alterations on 1p or 19q are infrequent in pediatric compared to adult OGs and are virtually absent in OGs presenting in the first decade of life. Compared to adults therefore, different genetic pathways are likely involved in the pathogenesis of most pediatric OGs. Genomic screening on a larger series is clearly indicated to delineate the unique molecular characteristics of these rare pediatric tumors. PMID- 12769193 TI - Anthony S. Fauci, MD, director, National Institute of Allergy and Infectious Diseases. PMID- 12769194 TI - Dr. Douglas Richman wins VA's Middleton Award. PMID- 12769195 TI - Intercellular adhesion molecule 1 is important for the development of severe experimental malaria but is not required for leukocyte adhesion in the brain. AB - Plasmodium berghei-infected mice, a well-recognized model of experimental cerebral malaria (ECM), exhibit a systemic inflammatory response. Most investigators hypothesize that leukocytes bind to endothelial cells via intercellular adhesion molecule 1 (ICAM-1), which causes endothelial damage, increased microvascular permeability, and, ultimately, death. ICAM-1-deficient mice on an ECM-susceptible C57BL/6 background were significantly (p = .04) protected from P. berghei mortality compared with ICAM-1 intact controls. ICAM-1 expression assessed by the dual radiolabeled monoclonal antibody technique was increased in the brain and lung in C57BL/6 mice on day 6 of P. berghei infection compared with uninfected controls (5.3-fold, p = .0003 for brain and 1.8-fold, p = .04 for lung). The increase in ICAM-1 expression coincided with significant (p < .05) increases in microvascular permeability in the brain and lung. In contrast to the hypothesized role for ICAM-1, in vivo analysis by intravital microscopy of leukocyte rolling and adhesion in brain microvasculature of mice revealed markedly increased levels of leukocyte rolling and adhesion in ICAM-1-deficient mice on day 6 of P. berghei infection compared with uninfected controls. In addition, ICAM-1 expression and microvascular permeability were increased in infected ECM-resistant BALB/c mice compared with uninfected BALB/c controls. These results collectively indicate that although ICAM-1 contributes to the mortality of experimental malaria, it is not sufficient for the development of severe experimental malaria. In addition, ICAM-1 expressed on the endothelium or on leukocytes is not required for leukocyte rolling or adhesion to the brain microvasculature of mice during P. berghei malaria. Leukocyte rolling and adhesion in the brain vasculature during P. berghei malaria use different ligands than observed during inflammation in other vascular beds. PMID- 12769196 TI - Patient education strategies to improve pneumococcal vaccination rates: randomized trial. AB - BACKGROUND: The pneumococcal vaccine is widely underused. Patient education is one mechanism not widely explored for increasing vaccination rates. OBJECTIVE: To evaluate the effects of a culturally appropriate patient education videotape on pneumococcal vaccination rates among the clinic population of an inner-city public hospital. METHODS: Randomized, controlled trial comparing (1) a videotape brochure group who both viewed the videotape and received a low-literacy brochure, (2) a videotape only group, and (3) a control group. RESULTS: Of 2,962 charts reviewed, 558 patients were randomized. The study population was 94% black, 73% female, and elderly (mean age 63.0 years) and 64% had less than a high school education. Patients in the videotape-brochure group were 2.5 (1.8, 3.5 95% CI) times more likely to discuss the vaccine with their physician (p < .001) and 3.5 (1.9, 6.5 95% CI) times more likely to receive the vaccine (p < .001) than the control group. The videotape-brochure group was 1.6 (1.2, 2.1 95% CI) times more likely to discuss the vaccine (p < .001) and 2.3 (1.4, 3.8 95% CI) times more likely to receive the vaccine (p = .002) than the video only group. Patients in the video only group were 1.6 (1.1, 2.3 95% CI) times more likely to discuss the vaccine with their physician than the control group (p = .041) but were not more likely to receive the vaccine. CONCLUSION: A culturally appropriate videotape along with a low-literacy brochure significantly increased pneumococcal vaccination rates and physician-patient discussion about the vaccine. These significant outcomes were not observed with use of videotape alone and were likely attributable to the effect of the brochure. We recommend that patient education initiatives to increase vaccination rates not focus solely on audiovisual media. PMID- 12769198 TI - The Genetic Sonogram Scoring Index. AB - This article describes the individual sonographic markers used in the Genetic Sonogram Scoring Index. The importance of the clustering of markers forms the basis of the scoring index, such that individual markers are assigned point values based on their sensitivity and specificity in the detection of Down syndrome. The points acquired by each fetus are tabulated into a final "score." The performance of the scoring index in the detection of aneuploidy is presented. The clinical application of the scoring index for the detection of chromosomally abnormal fetuses in patients both at high- and low-risk for aneuploidy is discussed. PMID- 12769197 TI - Impact of medical student research in the development of physician-scientists. AB - CONTEXT: A decline in the number of physician-scientists has been identified in the United States for at least two decades. Although many mechanisms have been proposed to reverse this trend, most of these have concentrated on MD/PhD programs, research in sub-specialty fellowships, and other approaches later in physician training. Few have emphasized early medical student research experiences as a contributing solution. OBJECTIVE: To determine the effect of a medical student research experience on career choices and attitudes about biomedical research. DESIGN, SETTING, AND PARTICIPANTS: We jointly report 25 years of experience with National Institutes of Health (NIH)-sponsored Medical Student Research Fellowship programs (MSRFs) at two colleges of medicine, the University of Tennessee Health Science Center and Vanderbilt University. In both programs, students work during the summer of their first or second year of medical school on a research project that is mentored by an established scientist and participate in a structured program (lectures, visiting professor). MAIN OUTCOME MEASURES: We gathered data using pre- and postresearch fellowship questionnaires to assess (a) quality of research experiences; (b) tabulation of productivity, that is, presentations, abstracts, publications, and awards; (c) long-term tracking of former program participants; (d) comparison of residency placements by medical student researchers; and (e) comments from former program participants on the effects of their students' research experiences on career choices. RESULTS: During this time, approximately 1,000 medical students participated in the two programs. Follow-up data (for short-term evaluations, 96 132 respondents with a response rate > 82%; for long-term evaluations, 88-118 respondents with a response rate > 29-33%) strongly suggest (a) interest in an academic career increased, (b) one-third to half of former student respondents considered themselves to be in academic medicine, (c) the vast majority of students conducted additional research after their medical student research experience, and (d) a large number of students were currently doing research or had published or presented their work at scientific meetings. CONCLUSIONS: Over two decades of experience with NIH-sponsored medical student research programs at two medical schools strongly support the ability of these programs to interest medical students in research and academic careers. MSRFs should be included in strategies to reverse the decline in the number of physician-scientists. PMID- 12769199 TI - Use of genetic sonography for adjusting the risk for fetal Down syndrome. AB - Systematic evaluation of ultrasound findings known to be associated with trisomy 21, at an appropriate gestational age, has been referred to as a genetic sonogram. A number of high-risk centers performing genetic sonography have reported detection of ultrasound abnormalities in the majority of fetuses with fetal Down syndrome. However, nonspecific markers are more commonly observed than structural abnormalities, which are detected in less than 20% of cases in a nonselected population. Also, the actual sensitivity of a genetic sonogram will depend on various factors including the markers sought, gestational age, reasons for referral, and of course the quality of the ultrasound. Appropriate use of a genetic sonogram can help to modify the risk of fetal Down syndrome by decreasing the risk when the ultrasound is normal, or increasing the risk when specific ultrasound markers are detected. The postultrasound risk can be estimated by applying specific likelihood ratios, reflecting the strength of individual markers, with the a priori risk based on maternal age alone, or combined with biochemical markers when known. We review this approach of age-adjusted ultrasound risk assessment for fetal Down syndrome and illustrate how the risk can be estimated. Individual sonographic markers are also discussed. PMID- 12769200 TI - Combined serum and ultrasound screening for detection of fetal aneuploidy. AB - Experience gathered over the last decade from high-risk centers provide strong evidence that mid-trimester sonographic markers are sensitive for Down syndrome prediction. More recent data indicate that combining mid trimester sonography with traditional serum markers significantly improves diagnostic accuracy over either group of markers by themselves. PMID- 12769201 TI - The use of genetic sonography to reduce the need for amniocentesis in women at high-risk for Down syndrome. AB - Much information has been published regarding the use of second-trimester genetic sonography for the prenatal detection of Down syndrome by examining multiple aneuploidy markers. Among high-risk mothers (advanced maternal age, abnormal triple screen, or both), while many undoubtedly will choose to have invasive testing as a first option, others will instead use the information derived from genetic sonography to obtain an adjusted risk for Down syndrome to guide their decision about genetic amniocentesis. Accordingly, it is imperative that these patients have accurate and detailed counseling regarding their degree of risk reduction when the genetic sonogram is normal. This article reviews the use of second trimester genetic sonography in reducing the need for amniocentesis in the high-risk patient. At our institution, in high-risk patients when the genetic ultrasound is normal, the amniocentesis rate has been only 3%. We have found that genetic sonography is a patient-driven service, and that the information obtained at the time of ultrasound is an important component of the patient's decision of whether or not to proceed with invasive testing. PMID- 12769202 TI - The role of fetal echocardiography in genetic sonography. AB - Genetic sonography identifies between 60% and 93% of fetuses with trisomy 21. One of the reasons for the variation in sensitivity is because of the under-detection of congenital heart defects. Although congenital heart defects are present in 56% of second trimester fetuses and 44% of newborns with trisomy 21, most studies evaluating second-trimester fetuses at risk for trisomy 21 detect less than 10% of heart malformations. This review discusses an approach that allows the fetal sonographer to incorporate fetal echocardiography, based upon the examiner's level of skill and experience, when evaluating the fetus at risk for trisomy 21. The cardiovascular examination consists of three levels. In the Level I examination only noncardiac markers are evaluated for a detection rate of 60% and false-positive rate of 5.9%. The Level II examination incorporates the four chamber view with non-cardiac markers. If the examiner can identify atrial and/or ventricular chamber disproportion, then the sensitivity is increased to 75%, with a false-positive rate of 6.4%. The Level III examination utilizes grayscale and color Doppler ultrasound to evaluate the fetal heart. If the examiner can identify ventricular septal defects, atrioventricular septal defects, pericardial effusion, tricuspid regurgitation, and chamber disproportion, then the sensitivity of genetic sonography increases to 91% with a false-positive rate of 14%. This review includes Likelihood Ratios for each of the ultrasound markers so that the examiner can compute the risk for trisomy 21 for an individual patient. PMID- 12769203 TI - Is genetic ultrasound cost-effective? AB - During the past 10 years, investigators have reported studies examining the potential of second-trimester genetic sonography to identify fetuses at risk for trisomy 21. The consensus among most investigators is that genetic sonography offers an alternative to universal amniocentesis in high-risk women and lowers the loss rate of normal fetuses subjected to amniocentesis because of risk factors associated with advanced maternal age or abnormal maternal-serum screening. Although there is now consensus that genetic sonography may be a useful screening tool, there has been a paucity of data regarding its cost effectiveness. In this review, 3 studies are examined and cost-effectiveness of genetic sonography evaluated. The first study compared genetic sonography and universal amniocentesis and found that genetic sonography was cost-effective if the sensitivity is 75% or higher, resulted in a savings to the healthcare system of 9%, and decreased the loss rate of normal fetuses following amniocentesis by 87%. The second study examined the use of genetic sonography in women less than 35 years of age who underwent maternal-serum triple-marker serum screening. Women who were screen negative but who were classified as moderate risk for trisomy 21 (risk 1:191 to 1:1,000) were offered genetic sonography. Amniocentesis was offered only if the genetic sonogram was abnormal. The study demonstrated that the use of genetic sonography in this group of patients increased the detection rate of trisomy 21, was cost effective, and was a safe procedure. The third study examined the use of genetic sonography in women 35 years of age and older who declined amniocentesis following second-trimester genetic counseling. Genetic sonography was offered to this group of patients followed by amniocentesis if an abnormal ultrasound finding was present. The data were analyzed for various acceptance rates of amniocentesis by the patient when informed of the ultrasound findings. Examination of the data demonstrated this approach increased the detection rate of trisomy 21, was cost-effective, and was a safe procedure. In conclusion, genetic sonography when applied in the above clinical settings is cost-effective, results in a higher detection rate of trisomy 21, and is safe procedure. PMID- 12769204 TI - Autoimmune pancreatitis presenting as a mass in the head of the pancreas: a diagnosis to differentiate from cancer. AB - We report a case of autoimmune pancreatitis presenting as a mass in the head of the pancreas that was successfully diagnosed without pancreaticoduodenectomy. The patient was a 64-year-old man who had no complaint. A routine physical checkup unexpectedly revealed mild diabetes and a low-echoic mass in the pancreatic head. The diagnosis was made by noting irregular narrowing of the main pancreatic duct, hypergammaglobulinemia, and increased immunoglobulin G levels. An open wedge biopsy of the mass was performed; this showed a marked fibrosis with lymphocyte- or macrophage-predominant inflammatory infiltrates. Immunohistochemical study revealed that the remnant acinar cells expressed Fas (CD95) ligand and not Fas. We review some of the literature and discuss various features and diagnostic clues of autoimmune pancreatitis. Awareness of this pathologic condition may prevent confusion with pancreatic malignancy and unnecessary surgery. PMID- 12769205 TI - Workforce patterns of rural surgeons in West Virginia. AB - Rural general surgeons perform a wide variety of procedures and have practices different from those of surgeons in larger communities. Because of this residents completing a classical general surgery training program may not be prepared for the rural setting. The 219 licensed physicians in West Virginia who list general surgery as their practice specialty with the State Board were surveyed to determine the nature of the rural surgery workforce and to examine the caseload of these surgeons. The majority of rural surgeons were satisfied with their current situation; however, 22 per cent stated that they would leave the practice of medicine if financially able. One-third of these surgeons regarded the rural setting as having an adverse impact on their practice. More than half of those surveyed stated that they would not encourage a young person to pursue a career in medicine. For one-third of rural surgeons general medicine was part of daily practice. The caseload varied by community size. Surgeons in communities of fewer than 10,000 people performed a lower percentage of general surgical procedures than surgeons in urban areas. They listed obstetric and gynecologic (9%), urologic (5%), otolaryngologic (9%), and orthopedic (4%) procedures as part of their regular cases. Endoscopic procedures comprise 17 to 24 per cent of total procedures regardless of community size. We conclude that resident surgeons planning to pursue a career in rural general surgery should broaden their operative and general medicine experiences to meet the needs of the communities in which they will serve. PMID- 12769206 TI - Prognostic indices in breast cancer are related to race. AB - African-American (AA) women have a higher mortality from breast cancer than Caucasians (C). This may be attributed to stage of disease at presentation, but specific prognostic factors are not well identified. We sought to identify prognostic factors in our database of early-stage (stage I and II) breast cancer from 1990 to 1999. There were 153 tumors in 150 AA women and 773 tumors in 760 C women. Prognostic factors are listed according to race with relative risk (RR) and 95 per cent confidence intervals. AA women presented significantly more often than C women under the age of 50 years (RR = 1.8) with palpable disease (RR = 1.3), higher-grade tumors (RR = 1.5), more estrogen receptor-negative disease (RR = 1.7), more progesterone receptor-negative disease (RR = 1.4), higher proliferation indices (RR = 1.9), and more lymph node-positive disease (RR = 1.6). Many of these adverse prognostic features persisted in "good" prognostic groups, i.e., those women over the age of 50 years with tumors <20 mm and having node-negative disease. We conclude that prognostic factors are related to race with AA women presenting at an earlier age and more often with palpable disease. More importantly AA women presented significantly more often with higher-grade tumors, hormone receptor-negative tumors, higher proliferation indices, and node positive disease. These findings may explain a higher breast cancer mortality in AA women. PMID- 12769207 TI - Laparoscopic adrenalectomy and splenectomy are safe and reduce hospital stay and charges. AB - The proposed benefits of laparoscopy for certain surgical procedures have been decreased post-operative pain and hospital stay balanced against the proposed deficits of increased costs. We have reviewed our data to evaluate factors associated with patient, procedure, and hospital charges for patients undergoing open versus laparoscopic adrenalectomy and splenectomy during the same time period. Eighty-seven patients underwent adrenalectomy (n = 47) or splenectomy (n = 40) from October 30, 1995 to June 6, 2001 and were retrospectively reviewed. Patient and operative factors were analyzed by intent to treat; the major endpoints were operating room (OR) time in minutes, blood loss in cm3, length of hospital stay in days, and charges broken down by anesthesia/operation [OR/recovery room (RR)] and total charges in dollars x 1000. Comparisons of means were analyzed by unpaired t test; data are presented as mean +/- SEM, and significance is defined as P < 0.05. Median age of the group was 47 years (range 20-77). Forty-five patients underwent a laparoscopic approach of which two were converted to open (4%) as compared with 42 undergoing an open operation; one patient from each group was excluded from outcome analyses because of prolonged hospitalization (>3 weeks). Operative mortality of the whole group was one per cent. There were no differences between the groups with respect to age, gender, or comorbidity. The laparoscopic group had significantly longer operative times and OR/RR charges. However, the length of hospital stay and the total charges for the patient undergoing a laparoscopic approach were significantly less (P < 0.05). We conclude that a laparoscopic approach for adrenalectomy or splenectomy can be accomplished in approximately 95 per cent of patients selected for this procedure. Despite prolonged OR time and increased OR/RR charges the laparoscopic procedures resulted in significantly decreased length of hospital stay and overall patient charges. Laparoscopy is a safe and cost-effective approach and should be strongly considered in patients requiring adrenalectomy or splenectomy. PMID- 12769208 TI - A prospective evaluation of a bedside technique for placement of inferior vena cava filters: accuracy and limitations of intravascular ultrasound. AB - Our objective was to evaluate the safety and accuracy of a bedside technique for placing vena cava filters with intravascular ultrasonography. We conducted a prospective case series of 36 patients requiring prophylactic vena cava filter placement. Intravascular ultrasound (IVUS) was used to assess the best location for inferior vena cava filter placement. Location of best filter placement was compared with a bedside technique using a fluoroscopic examination. Thirty-six patients underwent an IVUS examination. The 21 men and 15 women had a mean age of 51 years. Our bedside placement technique was successfully performed in 34 patients. Two patients did not undergo a bedside technique because of inadequate imaging. The difference between best filter placement and bedside technique was 1.22 +/- 1.24 cm. In three patients our bedside technique differed from best filter placement by more than 3 cm. Excluding these three patients the difference between best filter placement and bedside technique was 0.92 +/- 0.79 cm. This bedside IVUS technique for placement of inferior vena cava filters is established as safe and accurate; however, knowledge of the limitations involving the technique is important. PMID- 12769209 TI - Genetic testing for hereditary nonpolyposis colorectal cancer. AB - Approximately 80 per cent of patients with colorectal cancer have sporadic disease whereas the remaining 20 per cent seem to have a genetic component. Hereditary nonpolyposis colorectal cancer (HNPCC) is the most common autosomal dominant hereditary syndrome predisposing to colorectal cancer. Various methods have been described to screen for HNPCC and to directly test for mismatch repair gene mutations. This study evaluates the initial results of 1) microsatellite instability (MSI) and immunohistochemistry (IHC) staining of tumors and 2) genetic sequencing for mismatch repair gene mutations in patients suspected to have HNPCC. Appropriate patients for HNPCC testing were identified through a high risk colorectal cancer clinic. Of those patients screened only those who met Amsterdam criteria (AC) for HNPCC or were young age onset (YAO) (<40 years of age) were eligible for testing. The tumors underwent testing for MSI and had IHC performed in those patients with available tumor specimens. MSI was performed on the five markers approved by the NIH consensus conference. MSI-High (MSI-H) was defined as two or more markers being unstable. IHC was done with commercially available stains for MLH1 and MSH2. All patients had sequencing of the MLH1 and MSH2 genes performed to search for mutations by a commercial laboratory. Genetic counseling was provided and written informed consent was obtained. Fourteen patients were part of kindreds that met the AC. An additional 10 patients were <40 years of age at diagnosis of colorectal cancer but lacked any family history. Testing for MSI and IHC was performed on those available tissue blocks. Of the AC patients five had MSH2 mutations and two had MLH1 variants. Of the five with MSH2 mutations three of four had MSI-H tumors and all four had loss of expression of MSH2 on IHC. Of the MLH1 variants only one had MSI-H tumor and lacked expression of MLH-1 on IHC. Of those patients with no mutation identified three of six had MSI-H tumors. For those patients YAO no genetic mutations were identified. Two of the seven had MSI-H tumors. Genetic testing for HNPCC even in those patients fulfilling the rigid AC yielded mutations in only five of 14 patients with variants of unknown significance being found in an additional two patients. Only one MSH2 variant of unknown significance was identified in the 10 YAO patients, which would suggest that screening in this group of patients with MSI and/or IHC would be appropriate. PMID- 12769211 TI - Papillary carcinoma of the breast: characteristics and classification. AB - Papillary carcinoma is a rare but perplexing entity. The varied histopathological appearances make accurately diagnosing this tumor a difficult task. This review is a brief overview of the basic findings and a general classification of papillary carcinoma of the breast. Through an extensive literature review we have attempted to put forth a basic scheme of how papillary carcinoma is categorized with accepted treatment guidelines and protocol. A working knowledge of this tumor will facilitate a better understanding of its diagnosis and treatment with a reduction in overall morbidity. PMID- 12769210 TI - Impact of hypercalcemia and parathyroid hormone level on the sensitivity of preoperative sestamibi scanning for primary hyperparathyroidism. AB - Technetium 99m sestamibi scanning (MIBI) can direct unilateral parathyroidectomy. However, the clinical application remains variable with sensitivities ranging from 55 to 100 per cent. We examined whether patient factors including serum calcium (Ca) and parathyroid hormone (PTH) levels impact the sensitivity of MIBI. We completed a retrospective review of 102 patients with primary hyperparathyroidism and mild hypercalcemia who underwent preoperative MIBI. All patients underwent bilateral neck explorations with abnormalities confirmed by histopathology. MIBI sensitivity was correlated with preoperative Ca and PTH levels using univariate and logistic regression analysis. The mean preoperative Ca was 11.0 mg/dL and the mean PTH was 158 pg/mL. More than 95 per cent of patients with Ca greater than 11.3 mg/dL had a positive scan as compared with 60 per cent of those with lesser values (P = 0.0024). Similarly a serum PTH level greater than 160 pg/mL correlated with positive scans in 93 per cent as opposed to 57 per cent in those with lower levels (P = 0.006). Using a scan-directed approach 65 of 74 patients would have undergone unilateral exploration; this would yield a 7.7 per cent operative failure rate because of contralateral multigland disease. Lower Ca and PTH levels seem to correlate with reduced sensitivity of MIBI. Increasing acceptance of surgery for hyperparathyroidism with minimal hypercalcemia may make MIBI less attractive without ancillary diagnostic measures such as rapid parathormone assays. PMID- 12769212 TI - Annular pancreas in the adult: two case reports and review of more than a century of literature. AB - This paper brings the diagnosis and treatment of this interesting yet infrequent condition to contemporary standards through an assessment of past literature and a re-evaluation of the problem in view of technological advances. Two cases of annular pancreas are cited with a review of the last 183 years of literature. Only 100 cases, however, were reported in enough detail to examine outcomes in general and to arrive at a reasonable conclusion in terms of recommendations for diagnosis and operative intervention in adult annular pancreas. Two observations are brought to light. Despite all present diagnostic tools including endoscopic retrograde cholangiopancreatography diagnosis at best is made in only 60 per cent of patients preoperatively. Intraoperative expertise remains the best diagnostic modality available to date. Enteroenterostomy seems to be the intervention of choice for a multitude of anatomic and physiologic reasons and with a wide array of surgical options available when additional factors need to be addressed. PMID- 12769213 TI - Hemodynamic monitoring in the elderly undergoing elective colon resection for cancer. AB - Controversy surrounds the use of pulmonary artery catheters (PACs). We evaluated the influence of preoperative hemodynamic monitoring and optimization on the outcome in elderly patients undergoing elective resection for colon cancer. We performed a retrospective analysis of all elderly patients (age > 65 years) who had undergone elective colon resection during 1985 to 1995. Sixty patients had preoperative insertion of PAC; 217 patients were managed without PAC. Charts were reviewed for Goldman's cardiac risk index (CRI), preoperative risk factors, and hospital mortality. On the basis of CRI the patients were divided into two groups (< 10 and > or = 10). There was no significant difference between PAC or no-PAC patients for age, previous myocardial infarction, congestive heart failure, hypertension, chronic obstructive pulmonary disease, renal insufficiency, hemoglobin, and albumin. There were 12 deaths (4.3%). CRI, which was a significant predictor of mortality in the no-PAC group (2.2% mortality for CRI < 10 vs 15.8% for CRI > or = 10; P < 0.001), was insignificant in the PAC group (2.5% mortality for CRI < 10 vs 5% for CRI > or = 10, P = not significant). Although preoperative optimization using PAC was not beneficial in the low-CRI group it resulted in a threefold reduction in mortality (5% vs 15.8%) in the high CRI group. We conclude that preoperative optimization of cardiovascular function using a PAC is only beneficial in reducing mortality in high-risk (CRI > or = 10) elderly patients undergoing elective colon resection. PMID- 12769214 TI - Hepatic radiofrequency ablation of metastatic ovarian granulosa cell tumors. AB - Intrahepatic recurrences of a granulosa cell tumor of the ovary after primary resection has traditionally been considered a relative contraindication to surgical management. Improvements in ablative technologies such as radiofrequency ablation (RFA) offer the surgeon additional alternatives in the management of selected intrahepatic tumors. We present a case report of an intrahepatic recurrence of a metastatic ovarian granulosa cell tumor 6 months after primary resection. The patient received RFA of the intrahepatic lesions and the patient remains free of detectable disease 14 months later. A review of the literature is presented. This is the first known report of the use of RFA for intrahepatic recurrence of a metastatic ovarian granulosa cell tumor. In selected cases of metastatic ovarian granulosa cell tumors to the liver RFA may increase the percentage of patients considered surgically treatable. PMID- 12769215 TI - Effect of increasing the surface sampled by imprint cytology on the intraoperative assessment of axillary sentinel lymph nodes in breast cancer patients. AB - As axillary sentinel nodes predict the nodal status and may allow dissection of the axilla on a selective basis we assessed the effects of increasing the surface sampled during intraoperative imprint cytology. Sentinel nodes from 110 patients identified with Patent blue and/or the high radioactivity due to the uptake of 99m-Tc-labeled colloidal albumin were analyzed via hematoxylin and eosin-stained touch preparations. Imprint cytology was performed either on bisected nodes (Protocol One; n = 55) or on sentinel nodes sliced into multiple pieces at 2- to 3-mm intervals (Protocol Two; n = 55). The sentinel nodes were submitted in toto to permanent step sectioning and immunostaining for cytokeratins. There were equal numbers of patients with involved nodes in the two groups assessed. With Protocols One and Two the imprints had sensitivities of 52 and 61 per cent, negative predictive values of 74 and 78 per cent, and false negative rates of 47 and 39 per cent, respectively. No macrometastasis missed by Protocol Two was absent from the surface sampled. These data suggest that increasing the surface sampled improves the proportion of involved sentinel nodes detected intraoperatively by imprint cytology, but a number of metastatic nodes still remain undetected by this method. The sampling of multiple surfaces is encouraged for a more accurate intraoperative assessment. PMID- 12769216 TI - Use of porcine small intestinal submucosa in Ehlers-Danlos syndrome Type IV. AB - Ehlers-Danlos syndrome (EDS) Type IV results in inadequate production of type III collagen and can lead to devastating vascular, gastrointestinal, and uterine complications. A case is presented of spontaneous colonic perforation in a patient with EDS Type IV complicated by multiple intestinal fistulas managed with porcine small intestinal submucosa. Early success with this technique would suggest additional applications in a variety of complicated gastrointestinal problems. PMID- 12769217 TI - Laparoscopic adrenalectomy: a worthwhile procedure performed in a general surgery department. AB - A laparoscopic procedure is considered the treatment of choice for adrenalectomy. We report the experience of a nonreferring unit for adrenal pathology; we have evaluated its safety and feasibility in a series of 40 patients. From 1994 to 2001, forty consecutive patients underwent laparoscopic adrenalectomy, 37 with transperitoneal and 3 with retroperitoneal approach. The mean operative time was 129 +/- 51.7 minutes (range 60-300): 107 +/- 29 minutes (range 60-100) for the right-sided transperitoneal adrenalectomy and 144 +/- 62 minutes (range 90-300) for the left-sided transperitoneal adrenalectomy. The mean intraoperative blood loss was 90 mL (range 40-200). The procedure laparoscopic was converted to open in one case for the presence of a voluminous angiolipoma arising from the retroperitoneal fat strictly adherent to the adrenal gland. The postoperative morbidity rate was 5.1 per cent. Pain medication was required for a mean period of 1.6 +/- 0.6 days (range 1-3). The patients were able to resume solid food after an average time of 1.8 +/- 0.7 days (range 1-4). Postoperative hospital stay was 3 +/- 1.4 days (range 2-8). We believe that laparoscopic adrenalectomy is safe and effective in removing benign functioning or nonfunctioning adrenal masses and also in a general surgery department. PMID- 12769218 TI - Subcuticular closure versus Dermabond: a prospective randomized trial. AB - 2-Octylcyanoacrylate tissue adhesive (Dermabond, Ethicon, Inc, Somerville, NJ) is being used successfully for closure of minor lacerations. To date, however, there have been no studies evaluating its use in the operating room for surgical incisions. We conducted a prospective randomized trial to compare the closure of inguinal herniorrhaphy incisions using 2-octylcyanoacrylate tissue adhesive (Dermabond) with closures using 4-0 Monocryl (Ethicon, Inc) in a running subcuticular closure. A total of 46 incisions were randomized at the time of closure. Of these incisions 24 were randomized to Dermabond closure (TA) and 22 were randomized to subcuticular closure (SC). Performance measures included: time for closure, wound complications, and cosmesis. Cosmesis was evaluated by blinded evaluation of photographs of the incisions taken 4 weeks after surgery. Closure times for the TA group were faster than in the SC group (mean of 155 vs 286 seconds; P < 0.001). Wound complications were higher in the TA group (P = 0.045). Cosmesis was also felt to be better in the SC group with a score of 4.2 versus 3.88, but this did not reach statistical significance. Although the use of Dermabond did result in faster wound cultures it also resulted in an increase in wound complications. The difference in mean cosmetic score for each group was not statistically significant but trended toward better scores in the SC group. Based on these findings we do not feel Dermabond is an acceptable alternative to subcuticular suture closure in inguinal herniorrhaphy incisions. PMID- 12769219 TI - Training fourth-year medical students in critical invasive skills improves subsequent patient safety. AB - Complications after procedures performed by residents are thought to occur most often early in the first postgraduate year (PGY-1). We evaluated the number of pneumothoraces (PTXs) caused by central venous line insertion (CVLI) by two groups of PGY-1 residents in both the first 3 months of residency and the entire year from 1996 through 2000 to determine the impact of CVLI training on PTX. From 1996 through 1998 fourth-year medical students had no specific training in CVLI and learned on the job as residents. Starting with the Class of 1999 we replaced this approach with a structured program in CVLI. Didactic sessions detailing anatomy and technique were followed by skill performance in a fresh cadaver model. Students performed skills initially under the direct supervision of a faculty member, who provided immediate feedback. Videotapes of this performance were reviewed with the students by both surgeons and kinesiologists to correct deficits before repeat sessions. Skills were repeated until competence was attained. Graduating students have made up greater than 90 per cent of our two hospitals' PGY-1 residents since 1996. Because these residents are responsible for CVLI we are able to obtain performance follow-up in actual clinical settings. We obtained the number of PTXs caused by CVLI for the years 1996 through 2000 as well as for the first 3 months of each academic year (July through September) to determine the impact of our program on this serious complication. The number of PTXs during the first 3 months of 1996-1998 remained stable. After the introduction of our teaching program the number decreased significantly in the years 1999-2000 when compared with 1996-1998 (P = 0.004, t test). The overall yearly decrease for 1999 versus 1996-1998 approached significance (P = 0.06). The introduction of a structured teaching program of CVLI skills appears to have a positive impact in reducing morbidity of PTX. The greatest impact occurs within the first 3 months of the new PGY-1 academic year. PMID- 12769220 TI - The longest survivor and first potential cure of an advanced cholangiocarcinoma by ex vivo resection and autotransplantation: a case report and review of the literature. AB - Bismuth type IV hilar cholangiocarcinoma (CC) carries a poor prognosis; however, ex vivo liver resection and autotransplantation (Atx) is theoretically a treatment option. There are only five previously reported cases of this procedure for hilar CC in the English literature, and most of them died early in the postoperative period. The only reported survivor died of tumor recurrence at 13 months. We are reporting a patient who has survived for 17 months without any sign of tumor recurrence. This probably represents the world's first cure for CC using this technique. This patient is a 26-year-old woman with a Bismuth Type IV CC. Portal vein involvement at the confluence was shown on angiogram, and in situ resection was deemed impossible. Ex vivo resection of segments five, six, seven, eight, and part of segment four was performed followed by a partial liver Atx. The pathology specimen demonstrated CC with clear margins. MRI and CT examinations done over the subsequent 17 months showed no evidence of recurrence. In conclusion ex vivo liver resection and Atx can be a viable option for cure among highly selected patients with CC. PMID- 12769221 TI - Presentation and treatment of venomous snakebites at a northern academic medical center. AB - Poisonous snakebites are relatively rare in the United States. The incidence of venomous snakebites is comparatively high in the southern states compared with the northern states and reports of these accidents from northern states is particularly uncommon. We report the experience with treatment of venomous snakebites at the University of Michigan over a 25-year period from 1976 to 2001. Six cases were identified and are described in detail. All patients were male and all were bitten in the upper extremity by pit vipers. One patient suffered a moderate envenomation and was treated with antivenin. Four other cases of mild envenomation occurred and two of these cases required antivenin therapy. One case was considered to represent a "dry" bite and required only 24-hour observation. There were no severe envenomations and no mortalities. On short-term follow-up all patients recovered without sequelae. This report demonstrates that venomous snakebites can be treated effectively at low-volume centers. PMID- 12769222 TI - Re: The diagnostic dilemma of diaphragm injury. PMID- 12769223 TI - In vivo gene transfer to dissect neuronal mechanisms regulating cardiorespiratory function. AB - This lecture reviews recent information from our laboratory regarding brainstem mechanisms regulating the arterial baroreceptor reflex. Our long-term goal is to understand some of the mechanisms involved in the etiology of essential hypertension. Our hypothesis is that this problem may arise, in part, because of changes within brainstem circuits controlling arterial pressure, and in particular to occlusion of baroreceptive information at the level of the primary afferent relay within the brainstem. Although it is established that baroreceptors provide a mechanism for short-term regulation of arterial pressure, there is convincing evidence that they also play a role in its long-term control (see Thrasher 2002, for an example). It follows that dysfunction of this reflex circuit could contribute to high blood pressure levels. Here, we discuss the central actions of angiotensin II on the baroreceptor reflex circuitry within the nucleus of the solitary tract (NTS) for arterial pressure control. Our findings have led us to hypothesize a novel form of intercellular communication within the NTS, one of vascular-neuronal signaling. PMID- 12769224 TI - Development of the brain: a vital role for cerebrospinal fluid. AB - There has been considerable recent progress in understanding the processes involved in brain development. An analysis of a number of neurological conditions, together with our studies of the hydrocephalic Texas (H-Tx) rat, presents an important role for cerebrospinal fluid (CSF) in the developmental process. The fluid flow is essentially one-way and the location of the choroid plexuses in the lateral, third, and fourth ventricles allows for the possibility of new components being added to the fluid at these points. The role of the fourth ventricular CSF is particularly interesting since this is added to the fluid downstream of the cerebral hemisphere germinal epithelium (the main site of cortical cell proliferation and differentiation) and is destined for the basal cisterns and subarachnoid space suggesting different target cells to those within the ventricular system. Moreover, other sources of additions to the CSF exist, notably the subcommissural organ, which sits at the opening of the third ventricle into the cerebral aqueduct and is the source of Reisner's fibre, glycoproteins, and unknown soluble proteins. In this paper a model for the role of CSF is developed from studies of the development of the cortex of the H-Tx rat. We propose that CSF is vital in controlling development of the nervous system along the whole length of the neural tube and that the externalisation of CSF during development is essential for the formation of the layers of neurones in the cerebral cortex. PMID- 12769226 TI - Bidirectional communication between the pineal gland and the immune system. AB - The pineal gland is a vertebrate neuroendocrine organ converting environmental photoperiodic information into a biochemical message (melatonin) that subsequently regulates the activity of numerous target tissues after its release into the bloodstream. A phylogenetically conserved feature is increased melatonin synthesis during darkness, even though there are differences between mammals and birds in the regulation of rhythmic pinealocyte function. Membrane-bound melatonin receptors are found in many peripheral organs, including lymphoid glands and immune cells, from which melatonin receptor genes have been characterized and cloned. The expression of melatonin receptor genes within the immune system shows species and organ specificity. The pineal gland, via the rhythmical synthesis and release of melatonin, influences the development and function of the immune system, although the postreceptor signal transduction system is poorly understood. Circulating messages produced by activated immune cells are reciprocally perceived by the pineal gland and provide feedback for the regulation of pineal function. The pineal gland and the immune system are, therefore, reciprocally linked by bidirectional communication. PMID- 12769225 TI - Hormonally derived sex pheromones in fish: exogenous cues and signals from gonad to brain. AB - Living in a medium that can limit visual information but readily exposes the olfactory organ to hormonal compounds released by conspecifics, fish throughout their long evolutionary history have had both clear cause and ample opportunity to evolve olfactory responsiveness to these potentially important chemical cues (hormonal pheromones). Indeed, water-borne steroids, prostaglandins, and their metabolites are detected with great sensitivity and specificity by the olfactory organs of diverse fishes, and exert important effects on reproductive behavior and physiology in major taxa including carps (goldfish), catfishes, salmon, and gobies. Best understood are goldfish, where periovulatory females sequentially release a preovulatory steroid pheromone and a postovulatory prostaglandin pheromone that dramatically affect male behavior, physiology, and reproductive fitness. Although the diverse array of hormonal products released and detected by fish indicates clear potential for species-specific hormonal pheromones, olfactory recordings showing similar patterns of hormone detection among closely related species provide little evidence of selection for specificity. By demonstrating that the actions of sex hormones and related products are not limited to reproductive synchrony within the individual, the relatively recent discovery of hormonal pheromones has considerably expanded our understanding of fish reproductive function, while providing valuable model systems for future study of olfactory function and pheromone evolution. PMID- 12769227 TI - A neuroendocrine model for prolactin as the key mediator of seasonal breeding in birds under long- and short-day photoperiods. AB - Seasonal breeding is associated with sequential increases in plasma luteinizing hormone (LH) and prolactin in the short-day breeding emu, and in long-day breeding birds that terminate breeding by the development of reproductive photorefractoriness. A model of the avian neuroendocrine photoperiodic reproductive response is proposed, incorporating a role for prolactin, to account for neuroendocrine mechanisms controlling both long- and short-day breeding. The breeding season terminates after circulating concentrations of prolactin increase above a critical threshold to depress gonadotropin releasing hormone (GnRH) neuronal and gonadotrope (LH) activity. Subsequently, photorefractoriness develops for prolactin secretion and for LH secretion, independently of high plasma prolactin. The breeding season in the emu is advanced compared with long day breeders, because after photorefractiness for both LH and prolactin secretion is dissipated, plasma concentrations of both hormones increase to maximum values while days are still short. PMID- 12769228 TI - Sex steroid communication in the ring dove brain during courtship. AB - This review examines possible role of progesterone receptor (PR) and androgen receptor (AR) "cross-talk" in the expression of courtship behaviour in the ring dove (Streptopelia risoria). In doves, although androgen has been mostly associated with aggressive courtship behaviour and progesterone with the initiation of incubation, progesterone administration to courting birds terminates the aggressive component of courtship whilst having no effect on nesting behaviour. Recent results in doves have identified a high density of androgen receptor and progesterone receptor immunoreactivity (AR-ir and PR-ir) in the hypothalamus of both sexes in regions known to be directly involved in courtship and incubation behaviour. Nuclear AR-ir in courting birds is widespread throughout the brain. Nuclear PR-ir is only localized in discrete regions of the preoptic hypothalamus of both sexes. In the anterior and posterior hypothalamus of courting birds an increase number of AR-ir and PR-ir neurons colocalizes (70 90%) in the nucleus preopticus anterior (POA), nucleus preopticus medialis (POM), nucleus preopticus paraventricularis magnocellularis (PPM), nucleus hypothalami lateralis posterioris (PLH), and tuberal hypothalamus (Tu). A lower percentage of colocalization is seen in birds at other stages of the breeding cycle. The high percentage of AR-ir and PR-ir colocalization in the preoptic hypothalamus of courting doves supports previous reports involving progesterone acting in these brain regions to terminate the androgen-dependent aggressive courtship behaviour in male doves. The increase in PR-ir staining intensity in AR-ir neurons in courting birds suggests that this progesterone-dependent termination of aggressive courtship display in males occurs at the receptor level and may be orchestrated by central oestrogen. PMID- 12769229 TI - Growth hormone in the nervous system: autocrine or paracrine roles in retinal function? AB - Growth hormone (GH) is primarily produced in the pituitary gland, although GH gene expression also occurs in the central and autonomic nervous systems. GH immunoreactive proteins are abundant in the brain, spinal cord, and peripheral nerves. The appearance of GH in these tissues occurs prior to the ontogenic differentiation of the pituitary gland and prior to the presence of GH in systemic circulation. Neural GH is also present in neonates, juveniles, and adults and is independent of changes in pituitary GH secretion. Neural GH is therefore likely to have local roles in neural development or neural function, especially as GH receptors (GHRs) are widespread in the nervous system. In recent studies, GH mRNA and GH immunoreactive proteins have been identified in the neural retina of embryonic chicks. GH immunoreactivity is present in the optic cup of chick embryos at embryonic day (ED) 3 of the 21-d incubation period. It is widespread in the neural retina by ED 7 but also present in the nonpigmented retina, choroid, sclera, and cornea. This immunoreactivity is associated with proteins in the neural retina comparable in size with those in the adult pituitary gland, although it is primarily associated with 15-16 kDa moieties rather than with the full-length molecule of approximately 22 kDa. These small GH moieties may reflect proteolytic fragments of "monomer" GH and (or) the presence of different GH gene transcripts, since full-length and truncated GH cDNAs are present in retinal tissue extracts. The GH immunoreactivity in the retina persists throughout embryonic development but is not present in juvenile birds (after 6 weeks of age). This immunoreactivity is also associated with the presence of GH receptor (GHR) immunoreactivity and GHR mRNA in ocular tissues of chick embryos. The retina is thus an extrapituitary site of GH gene expression during early development and is probably an autocrine or paracrine site of GH action. The marked ontogenic pattern of GH immunoreactivity in the retina suggests hitherto unsuspected roles for GH in neurogenesis or ocular development. PMID- 12769230 TI - Evaluation of the antiulcerogenic activity of violacein and its modulation by the inclusion complexation with beta-cyclodextrin. AB - The effects of beta-cyclodextrin (betaCD) inclusion complexation on the ability of violacein to prevent gastric ulceration in mice were studied. Violacein-betaCD inclusion complexes were prepared in 1:1 and 1:2 molar ratios and analysed by differential scanning calorimetry and powder X-ray diffractometry. Violacein previously administered orally at 10 mg/kg significantly reduced indomethacin induced gastric lesions, as well as 100 mg/kg of cimetidine (positive control). However, betaCD complexation in both molar ratios significantly potentiated the protective action of violacein. In the HCl--ethanol-induced gastric ulcer model, violacein and the 1:2 inclusion complex (10 mg/kg, p.o.) inhibited gastric damage by almost 85%, whereas a 63% reduction was observed for the positive control, lansoprazole, at 30 mg/kg. In contrast, treatment with the 1:1 inclusion complex resulted in almost total disappearance of the antiulcer activity in this model. No significant changes in stress-induced gastric injury were found. In addition, the 1:2 inclusion complex improved the antilipoperoxidant activity of violacein in rat liver cells exposed to t-butyl hydroperoxide, whereas the 1:1 complex was less active than violacein. In summary, the 1:2 betaCD inclusion complex has gastroprotective properties similar to or higher than that of violacein. An increase in mucosal defensive mechanisms and protection against peroxidative damage might be involved. PMID- 12769231 TI - Effects of contrast media on erythrocyte aggregation during sedimentation. AB - To evaluate the effects of contrast media (CMs) on erythrocyte aggregation, we measured the erythrocyte sedimentation with Westergren method at 25 degrees C. CMs were diatrizoate (Urografin 76%) for ionic CM and iopamidol (Iopamiron 370) for nonionic CM. Swine red blood cells (RBCs) were suspended in autologous plasma containing diatrizoate (URO), iopamidol (IOP), and saline (SAL) at 6.7% w/w, as well as in plasma alone (PLA), at 40% of the hematocrit. Sigmoid sedimentation curves were fitted to the Puccini et al. (1977) equation, and the average number of RBCs per aggregate m was calculated by Stokes' law against the time t. According to the Murata-Secomb (1988) theory we estimated the collision rate K between two aggregates from dm/dt in the stationary phase during sedimentation. Corresponding to the maximal ESR, the dm/dt (in cells/s) was 0.52 in PLA, 0.09 in SAL, 0.06 in URO and 0.03 in IOP, so that K also decreased in proportion to dm/dt from 145 fL/s in PLA to 8 fL/s in IOP. Both the ionic and nonionic CMs tend to inhibit the RBC aggregation more than that in SAL; the latter iopamidol appears to be inhibitory more than the former diatrizoate in autologous plasma. PMID- 12769232 TI - Thyroid hormone-induced sulfate absorption in Aplysia californica gut is mediated by protein synthesis. AB - Mucosal membranes of foregut epithelia of Aplysia californica contain a sodium sulfate symporter that is stimulated by triiodothyronine. Actinomycin D, puromycin, or cycloheximide inhibited the triiodothyronine-stimulated sulfate absorption. It appears that thyroid hormone manifests its effects on sulfate absorption in the A. californica gut through protein synthesis. PMID- 12769233 TI - Phosphate absorption in Aplysia californica gut: glucocorticoid inhibition. AB - Apical membranes of Aplysia californica foregut epithelia contain a sodium phosphate symporter. Dexamethasone inhibited the absorptive activity of the sodium-phosphate symporter, whereas amiloride had no effect on the sodium phosphate symporter. It appears that glucocorticoids or their molluscan equivalent play a role in the overall regulation of phosphate homeostasis by the Aplysia californica gut. PMID- 12769234 TI - Censorship looming. PMID- 12769235 TI - Posttraumatic stress and functional impairment in Kenyan children following the 1998 American Embassy bombing. AB - This study examined a convenience sample of 562 Nairobi school children exposed to the 1998 bombing of the American Embassy in Nairobi, Kenya. Posttraumatic stress reactions to the bombing were related to posttraumatic stress reactions to other trauma and to peritraumatic reaction. Self-reported functional impairment was minimal. PMID- 12769236 TI - Gender, types of conflict, and individual differences in the use of violent and peaceful strategies among children who have and have not witnessed interparental violence. AB - Child witnesses' use of violent or peaceful strategies to resolve conflicts with peers were compared with strategies used by nonwitnesses. Stories depicting 5 types of peer conflicts were presented to each child, and the strategies used by the child to reach peaceful resolutions were scored. Findings include differences between witnesses and nonwitnesses, differences between genders, differences according to types of conflicts, and individual differences. PMID- 12769237 TI - Clinical algorithms as a tool for psychotherapy with Latino clients. AB - Clinical algorithms have the advantage of being able to integrate clinical, cultural, and environmental factors into a unified method of planning and implementing treatment. A model for practice is proposed that uses 3 algorithms as guides for conducting psychotherapy with Latino clients, the uses of which are illustrated in a single, ongoing case vignette. The algorithm format has the additional advantage of easily adapting itself for data gathering for research purposes. PMID- 12769238 TI - Intolerance and psychopathology: toward a general diagnosis for racism, sexism, and homophobia. AB - Racism, sexism, and homophobia do not fit into any current diagnostic category. The authors propose that those who engage in such behaviors display a form of psychopathology deserving of its own category. The common denominator seems to be intolerance. The authors explore the possibility of an intolerant personality disorder, outline likely symptoms, and suggest some possible treatment considerations. PMID- 12769239 TI - Community violence and sociomoral development: an African American cultural perspective. AB - This review considers the impact of exposure to community violence on sociomoral development within the African American community. Common sequelae of covictimization, as well as cultural experiences of the African American community, frame a discussion of the implications of covictimization for the development of moral reasoning. Recommendations for future research and intervention are provided. PMID- 12769240 TI - Intercountry adopted children as young adults--a Swedish cohort study. AB - In a national cohort study, the family and labor market situation, health problems, and education of 5,942 Swedish intercountry adoptees born between 1968 and 1975 were examined and compared with those of the general population, immigrants, and a siblings group--all age matched--in national registers from 1997 to 1999. Adoptees more often had psychiatric problems and were longtime recipients of social welfare. Level of education was on par with that of the general population but lower when adjusted for socioeconomic status. PMID- 12769241 TI - Symbolic, relational, and ideological signifiers of bias-motivated offenders: toward a strategy of assessment. AB - Developmental, ideological, and behavioral characteristics of 58 convicted hate crime offenders were examined. Ratings on the Psychopathy Checklist: Screening Version, HCR-20, and Bias Motivation Profile (BMP) were made via record review. Offense characteristics were rated on the Cornell Aggression Index and Cormier Lang Crime Index. Results indicated that offenders with higher BMP scores engaged in more instrumental (i.e., premeditated) aggression and targeted racial-ethnic minority victims. Significant within-group variation in the prominence of offender bias motivation on the BMP was observed. PMID- 12769242 TI - Clinical recovery in cognitive functioning and self-image among adolescents with major depressive disorder and conduct disorder during psychiatric inpatient care. AB - This prospective study examined psychosocial and cognitive functioning among adolescents with major depressive disorder and conduct disorder under comprehensive psychiatric inpatient care. Nonverbal and general cognitive performance, self-image, and overall psychosocial functioning improved in both groups. Nevertheless, more changes in self-image and family functioning were found among patients with major depressive disorder. Mental health services should still have the capability of offering long-term residential care for adolescents with the most severe mental disturbance. PMID- 12769243 TI - Cultural, self-esteem, and demographic correlates of perception of personal and group discrimination among East Asian immigrants. AB - Perception of personal and group discrimination and their cultural, self-esteem, and demographic correlates were examined in 170 East Asian immigrants. Many East Asian immigrants endorsed experiences of personal and group discrimination. Cultural, self-esteem, and demographic features were differentially associated with perception of personal discrimination versus perception of group discrimination. PMID- 12769244 TI - Quadricuspid aortic valves: a review. PMID- 12769245 TI - Are "paradoxical emboli' really paradoxical? PMID- 12769246 TI - Aldosterone antagonism and hypertension. PMID- 12769247 TI - Sustained use of nesiritide to aid in bridging to heart transplant. AB - Patients with end-stage heart failure awaiting heart transplant are often maintained on continuous intravenous inotropic therapy. However, this therapy alone is often inadequate for maintenance of appropriate pulmonary artery pressure and stable clinical course. Nesiritide, B-type natriuretic peptide, is a recently released intravenous vasodilator for short-term use in patients with decompensated heart failure. This report details experience in four patients in whom this agent was used to bridge to transplant for prolonged periods (11-35 days) with added clinical benefit and without obvious tolerance. This suggests that new strategies for pretransplant management may be needed. PMID- 12769248 TI - Pulsus paradoxus in cardiac tamponade: a pathophysiologic continuum. AB - Pulsus paradoxus has interested physicians for more than a century. Since McGregor's comprehensive New England Journal of Medicine article in 1979, there have been no updated reviews; accordingly, we review pulsus paradoxus based on the clinical and physiologic literature and personal experience. PMID- 12769249 TI - The prognostic value of QTc interval and QT dispersion following myocardial infarction in patients treated with or without dofetilide. AB - BACKGROUND: Acute myocardial infarction (MI) is associated with an increased risk of death, with a 1-year mortality close to 10% in patients discharged from hospital alive. During the first year following MI, close to 50% of deaths are assumed to be due to arrhythmic events. HYPOTHESIS: The study was undertaken to determine the interaction between dofetilide treatment and pretreatment QTc interval and QT dispersion regarding mortality in patients with left ventricular (LV) dysfunction and a recent MI. METHODS: The study population consisted of 894 patients with a recent MI and LV systolic dysfunction, who were randomized to receive dofetilide or placebo. The study was a substudy of the Danish Investigations of Arrhythmia and Mortality on Dofetilide-MI (DIAMOND-MI). RESULTS: During a minimum of 1-year follow-up, 261 (29%) patients died. Baseline QTc interval did not hold any prognostic value on mortality for placebo-treated patients. When pretreatment QTc interval was <429 ms, dofetilide resulted in a 45% reduction of mortality (hazard ratio 0.55, 95% confidence limits 0.34-0.88, p<0.02) compared with placebo. When QTc interval was >429 ms, dofetilide did not influence mortality significantly. This study revealed no statistically significant relation between QT dispersion, dofetilide treatment, and mortality. CONCLUSION: In patients with a recent MI, LV dysfunction, and a short baseline QTc interval, dofetilide is associated with significant survival benefit. This benefit is not seen with a longer QTc interval. QT dispersion is not a risk factor in this population. PMID- 12769251 TI - Opportunities for improvement in the diagnosis and treatment of heart failure. AB - BACKGROUND: Improved treatment of congestive heart failure (CHF) can slow disease progression, promote clinical stability, and prolong survival. HYPOTHESIS: Patterns in diagnostic test utilization and pharmacotherapy among patients with newly diagnosed heart failure may affect outcomes. METHODS: Claims data were analyzed from all diagnostic procedures and prescriptions from 1995 to 1998 in 3,353 patients with heart failure diagnosed within 1 year. Rates of diagnostic testing and categories of drugs prescribed were the main outcome measures. Demographic variables and type of provider were analyzed within a setting whose access to care was controlled. RESULTS: Rates of diagnostic testing with respect to basic, metabolic/endocrine, alternative diagnoses, underlying ischemia, and left ventricular function varied as a function of gender, age, race, and primary versus specialty care provider. Only 4.7% of patients underwent all diagnostics and treatments recommended in current guidelines. However, those patients (27.5%) who underwent an evaluation for ischemic heart disease and were prescribed vasodilators or beta blockers enjoyed the lowest crude mortality. CONCLUSIONS: There are multiple opportunities apparent to improve the initial diagnostic and therapeutic care of patients with heart failure. There appears to be an early survival benefit with respect to use of vasodilators and beta blockers within the first year of treatment. PMID- 12769250 TI - Significance of exercise-induced ST changes in leads aVR, V5, and V1. Discrimination of patients with single- or multivessel coronary artery disease. AB - BACKGROUND: It is known that exercise-induced ST-segment elevation in lead V1 (V1 E) detects left anterior descending (LAD) stenosis. It was also postulated that ST elevation in aVR and simultaneous ST depression in V5 (aVR-E + V5-D) is a marker of ischemia due to significant stenosis of the LAD in patients with single vessel disease. HYPOTHESIS: This study was undertaken to investigate the significance of the concomitant appearance of both electrocardiographic (ECG) ischemic markers, and of each of them alone during exercise, to detect either LAD stenosis as single-vessel coronary artery disease (CAD), or multivessel CAD involving LAD stenosis. METHODS: A total of 196 consecutive patients (152 men and 44 women, mean age 54 +/- 7 years) with at least one of these ECG markers, who underwent treadmill exercise testing with the Bruce protocol and coronary arteriography, were studied. RESULTS: Patients were divided into three groups. In Group A (83 patients with V1-E + aVR-E & V5-D), 93% of patients with single vessel disease had significant LAD stenosis (p<0.001), whereas 75% of patients with double-vessel disease had significant stenoses of the LAD and the left circumflex (LCx) coronary arteries (p<0.01). In Group B (97 patients with aVR-E & V5-D but without V1-E), 43% of patients with single-vessel disease had significant LAD stenosis (p<0.08), whereas 85% of patients with double-vessel disease had significant stenoses of the LAD and the right coronary artery (RCA) (p<0.01). In Group C (16 patients with only V1-E), 60% of patients with single vessel disease had significant LAD stenosis (p<0.05), whereas 75% of patients with double-vessel disease had significant LAD and LCx stenoses (p<0.05). CONCLUSIONS: The concomitant appearance of exercise-induced ST elevation in lead V1, ST elevation in lead aVR, and ST depression in lead V5, as well as the isolated appearance of ST elevation in lead V1 detect significant LAD stenosis as single-vessel disease, or significant stenoses of LAD and LCx arteries in patients with double-vessel disease, whereas the appearance of ST elevation in aVR & ST depression in V5 but without ST elevation in V1 correlates strongly with significant LAD and RCA stenoses and usually indicates double-vessel disease. PMID- 12769252 TI - Images in cardiology: Massive epicardial adipose tissue indicating severe visceral obesity. PMID- 12769253 TI - Isometric handgrip exercise during dobutamine-atropine stress echocardiography increases heart rate acceleration and decreases study duration and dobutamine and atropine dosage. AB - BACKGROUND: Dobutamine-atropine stress echocardiography (DASE) is an established test for the diagnosis and risk stratification of patients with coronary artery disease. Atropine use to attain target heart rate prolongs test time. HYPOTHESIS: The aim of this study was to assess the utility of isometric handgrip exercise (33% maximal voluntary contraction x 4 min) with DASE. METHODS: We prospectively evaluated 131 patients undergoing DASE randomized to handgrip exercise or no handgrip. Effect of handgrip exercise on endpoints: time to target heart rate (85% maximum predicted), recovery time, total test time, mean dobutamine and atropine dosage, and the number of ischemic responses were assessed. Effect of current beta-blocker medication use was also evaluated. RESULTS: Heart rate rose more quickly in the handgrip group. At 6-10 min (peak handgrip), mean heart rate rose 51 +/- 14 beats/min in the handgrip group compared with 38 +/- 18 beats/min in the no handgrip group (p < 0.0001). With handgrip, overall dobutamine study time was reduced by a mean of 4.3 min (16.4 +/- 6.9 vs. 20.7 +/- 8.4, p = 0.004) in all patients, and by a mean of 5.9 min in patients not on beta-blocker medication (p = 0.001). The handgrip group also had a lower mean dose of dobutamine (25.8 +/- 13.5 vs. 32.4 +/- 16.4 mg, p = 0.025). The mean atropine dose was also lower (0.2 +/- 0.4 vs. 0.4 +/- 0.5 mg, p = 0.04). Handgrip exercise, however, did not decrease endpoints in patients on beta-blocker medication. CONCLUSIONS: Use of isometric handgrip exercise with DASE decreases time to target heart rate, recovery time, overall study time, and mean dosage of dobutamine and atropine. In patients not on beta-blocker medication, handgrip exercise should be routinely incorporated into all DASE protocols. PMID- 12769255 TI - Images in cardiology: Infiltrating metastatic cardiac tumor. PMID- 12769254 TI - Guideline implementation in a multicenter study with an estimated 44% relative cardiovascular event risk reduction. AB - BACKGROUND: The extent of cardiovascular risk reduction by implementing coronary prevention guidelines needs to be documented in various population samples. HYPOTHESIS: This is a multicenter study to assess the impact of risk reduction in cardiovascular events upon implementation of coronary prevention guidelines in patients with or at high risk for coronary heart disease (CHD) in the setting of clinical practice. METHODS: Enrolled volunteers numbered 2,021. Inclusion criteria postulated a minimum of 20-40% cardiovascular event risk in the subsequent 10 years as estimated from the risk table of the European Society of Cardiology (ESC) Guidelines. The estimated CHD risk reduction was assessed in terms of the Framingham risk scores at baseline and at 12 months, computed from the data of each individual. Data of the compliant group (making up half of the initial participants) at the end of the study, along with absolute and relative risk reductions in the compliant group, were analyzed. RESULTS: Mean global risk burden was 25.9% at baseline, reduced through multilateral preventive measures in absolute terms by 9.4% at 6 months and by 11.7% at 12 months; the latter represents a relative risk reduction of 44%. Independent variables determining the (enhanced) reduction in risk level at the end of 12 months included (high) level of baseline risk, (high) degree of compliance with treatment, younger age, female gender, smoking, and (high) baseline triglyceride/high-density lipoprotein cholesterol (TC/HDL-C) ratio. While the relative reduction in patients with CHD amounted to 43%, a reduction of 46% (p<0.001) was obtained in the setting of primary prevention. Diabetes emerged as a factor modestly limiting the extent of risk reduction. While subjects without hypertension revealed a decline of coronary risk by merely 8.7%, those with hypertension showed a decline by 12.7% (p<0.001). Risk reductions were accompanied by a decrease of mean low-density lipoprotein cholesterol (LDL-C) level of 25.4%, a rise in mean HDL-C level of 5 mg/dl, a decrease in mean systolic blood pressure of 26 mmHg. Forty-five percent of smokers succeeded in discontinuing the habit. CONCLUSION: By implementing standard prevention guidelines in the Turkish population among 1,000 compliant high-risk men and women and among 1,000 patients with CHD, prevention of cardiovascular events could be expected in 117 persons in the subsequent 10 years. PMID- 12769256 TI - A. John Camm. PMID- 12769257 TI - Differential expression of connexin43 in foetal, adult and tumour-associated human brain endothelial cells. AB - Connexin43 (Cx43), the main protein constituting the gap junctions between astrocytes, has previously been demonstrated in endothelial cells of somatic vessels where the intercellular coupling that it provides plays a role in endothelial proliferation and migration. In this study, Cx43 expression was analysed in human brain microvascular endothelial cells of the cortical plate of 18-week foetal telencephalon, in adult cerebral cortex and glioma (astrocytomas). The study was carried out by immunocytochemistry utilizing a Cx43 monoclonal antibody and a polyclonal antibody anti-GLUT1 (glucose transporter isoform 1) to identify the endothelial cells and to localize Cx43. Endothelial Cx43 is differently expressed in the cortical plate, cerebral cortex and astrocytoma. Within the cortical plate and tumour, Cx43 is highly expressed in microvascular endothelial cells whereas it is virtually absent in the cerebral cortex microvessels. The high expression of the gap junction protein in developing brain, as well as in brain tumours, may be related to the growth status of the microvessels during brain and tumour angiogenesis. The lack of endothelial Cx43 in the cerebral cortex is in agreement with the characteristics of the mature brain endothelial cells that are sealed by tight junctions. In conclusion, the results indicate that endothelial Cx43 expression is developmentally regulated in the normal human brain and suggest that it is controlled by the microenvironment in both normal and tumour-related conditions. PMID- 12769258 TI - Subcellular distribution of cadmium and its cellular ligands in mussel digestive gland cells as revealed by combined autometallography and X-ray microprobe analysis. AB - Autometallography (AMG) and electron probe X-ray microanalysis (EPXMA) were applied in combination to determine the subcellular distribution of Cd and its subcellular ligands in the digestive gland cells of Cd-exposed mussels Mytilus galloprovincialis. Black silver deposits (BSD), which reveal the presence of metals when AMG is applied, were only localized in digestive cell lysosomes. Digestive cell cytoplasm and basophilic cells were devoid of BSD. EPXMA (static probe and X-ray mapping) indicated that Cd, S (possibly associated with metallothioneins or metallothionein-like proteins) and autometallographical Ag ions are co-localized within digestive cell lysosomes. In addition, Cd and S co occur in the absence of Ag in the cytosol of digestive cells. AMG does not reveal the presence of the Cd 'pool' strongly bound to cytosolic Cd-metallothionein complexes; only 'free' Cd or Cd supposedly loosely bound to (semi)digested metallothionein within lysosomes was revealed. The levels of lysosomal Cd were indirectly quantified by stereology as the volume density of BSD (V(v)BSD). Significantly higher values were recorded in Cd-exposed mussels compared with controls at all exposure times. However, V(v)BSD values were lower at days 7 and 21 than at day 1. This relative decrease in V(v)BSD reflected another (and confounding) response elicited by Cd-exposure in the digestive epithelium: the volume density of basophilic cells (V(v)BAS) increased significantly as exposure progressed. Due to this cell-type replacement, the net accumulative capacity of the digestive epithelium decreases at long exposure times. PMID- 12769259 TI - Heterogeneity in olfactory neurons in mouse revealed by differential expression of glycoconjugates. AB - Cell surface glycoconjugates have been implicated in the growth and guidance of subpopulations of primary olfactory axons. While subpopulations of primary olfactory neurons have been identified by differential expression of carbohydrates in the rat there are few reports of similar subpopulations in the mouse. We have examined the spatiotemporal expression pattern of glycoconjugates recognized by the lectin from Wisteria floribunda (WFA) in the mouse olfactory system. In the developing olfactory neuroepithelium lining the nasal cavity, WFA stained a subpopulation of primary olfactory neurons and the fascicles of axons projecting to the target tissue, the olfactory bulb. Within the developing olfactory bulb, WFA stained the synaptic neuropil of the glomerular and external plexiform layers. In adults, strong expression of WFA ligands was observed in second-order olfactory neurons as well as in neurons in several higher order olfactory processing centres in the brain. Similar, although distinct, staining of neurons in the olfactory pathway was detected with Dolichos biflorus agglutinin. These results demonstrate that unique subpopulations of olfactory neurons are chemically coded by the expression of glycoconjugates. The conserved expression of these carbohydrates across species suggests they play an important role in the functional organization of this region of the nervous system. PMID- 12769260 TI - Temporal and spatial expression of fibroblast growth factor receptor 4 isoforms in murine tissues. AB - Fibroblast growth factor receptors (FGFRs) undergo highly regulated spatial and temporal changes of expression during development. This study describes the use of quantitative reverse transcriptase-polymerase chain reaction and immunochemistry to assess the changes in expression of FGFR4 as compared to its FGFR4-17a and -17b isoforms in mouse tissues, from early embryogenesis through to adulthood. Compared to FGFR4, the expression of the isoforms is more restricted at all developmental stages tested. The reverse transcriptase-polymerase chain reaction demonstrated that FGFR4 is expressed in more tissue types than either of its isoforms: it was found predominantly in lung, liver, brain, skeletal muscle and kidney, whereas the FGFR4-17a form was detected in lung and skeletal muscle, and the FGFR4-17b form only in lung, liver, skeletal muscle and kidney. Immunohistochemistry confirmed strong FGFR4-17b expression in the postnatal lung. When combined, the results suggest that FGFR4 variants play important roles particularly in lung and skeletal muscle development. PMID- 12769261 TI - Pyronin Y as a fluorescent stain for paraffin sections. AB - Pyronin Y has long been used, in combination with other dyes such as Methyl Green, as a differential stain for nucleic acids in paraffin tissue sections. It also forms fluorescent complexes with double-stranded nucleic acids, especially RNA, enabling semi-quantitative analysis of cellular RNA in flow cytometry. However, the possibility of using pyronin Y as a fluorescent stain for paraffin tissue sections has rarely been investigated. We herein report that in sections stained with Methyl Green-pyronin Y, red blood cells, elastic fibre of blood vessels, zymogen granules of pancreatic acinar cells, surface membrane of heptocytes and kidney tubular cells showed strikingly strong green and/or red fluorescence, while the nuclei of cells appeared non-fluorescent. The use of confocal laser-scanning microscope greatly improved the resolution and selectivity of the fluorescent images. Staining with pyronin Y alone gave similar results in terms of fluorescence properties of the specimens. Pretreatment of paraffin sections with RNase significantly reduced cytoplasmic pyronin Y staining as judged by transmission light microscopy, but it had little effect on the fluorescence intensity of red blood cells, elastic fibres and zymogen granules. PMID- 12769262 TI - Small heat shock proteins expression in rat kidneys treated with cyclosporine A alone and combined with melatonin. AB - Small heat shock proteins (sHSPs) are cytoskeletal chaperones constitutively expressed in the normal kidney but enhanced with beneficial roles during adverse stimuli. Cyclosporine A is an immunosuppressive drug with major adverse side effect such as severe nephrotoxicity. Among possible mechanisms of cyclosporine A induced renal damage, oxidative stress and cytoskeletal damage have been suggested. Melatonin has been successfully used as antioxidant against many renal diseases. This in vivo study was performed to shed light on the protective effect of melatonin against cyclosporine A-induced renal alterations. We treated rats with cyclosporine A alone, or combined with melatonin, and with melatonin alone (as controls) for 40 days and analysed the renal abundance and distribution of two sHSPs, HSP25 and alpha B-crystallin. These data were correlated with the histopathological effects of the treatments. Cyclosporine A induced insoluble isoforms that moved to soluble fractions after melatonin coadministration as in controls. After cyclosporine A treatment, an intense signal for sHSPs was found within the glomeruli, nucleus and cytoplasm of cortical tubules, collecting ducts and vascular wall. After melatonin supply, the staining was faint, limited to the cytoplasm of cortical tubules, similar to controls. Both fibrosis and tubular alterations significantly decreased after melatonin coadministration. In conclusion, HSP25 and alpha B-crystallin are overexpressed in the rat kidney treated with cyclosporine A but are similar to controls after combined melatonin. This could be a consequence of the cytoprotective effect of melatonin in this nephrotoxic model so that a beneficial sHSPs response is unnecessary. PMID- 12769263 TI - Immunohistochemical localization of human kallikreins 6 and 10 in pancreatic islets. AB - Tissue kallikreins are thought to be present in the pancreatic islets of Langerhans and to aid in the conversion of proinsulin to insulin. In recent immunohistochemical studies, we observed strong staining of the newly identified human kallikreins 6 and 10 (hK6 and hK10) in the islets of Langerhans. Here, we examine hK6 and hK10 immunoexpression in different types of islet cells of the endocrine pancreas, in order to obtain clues for hK6 and hK10 function in these cells. Ten cases of normal pancreatic tissue, two cases of nesidioblastosis, five insulin-producing tumours and one case of multiple endocrine neoplasia 1 syndrome, containing an insulin-, a somatostatin- and several glucagon-producing tumours, as well as tiny foci of endocrine dysplasia with different predominance of the secreted hormones (mainly glucagon and pancreatic polypeptide) were included in the study. A streptavidin--biotin--peroxidase and an alkaline phosphatase protocol, as well as a sequential immunoenzymatic double staining method were performed, using specific antibodies against hK6, hK10, insulin, glucagon, somatostatin, pancreatic polypeptide, and serotonin. hK6 and hK10 immunoexpression was observed in the islets of Langerhans, including the pancreatic polypeptide-rich islets, in the normal pancreas. Scattered hK6 and hK10 positive cells were localized in relationship with pancreatic acinar cells. In the exocrine pancreas, a cytoplasmic and/or brush border hK6 and hK10 immunoexpression was observed in the median and small sized pancreatic ducts, while the acinar cells were negative. Foci of nesidioblastosis and endocrine dysplasia expressed both kallikreins. hK6 and hK10 were also strongly and diffusely expressed throughout all insulin-, glucagon- and somatostatin-producing tumours. The double staining method revealed co-localization of each hormone and hK6/hK10 respectively, in the same cellular population, in the normal as well as in the diseased pancreas. Our results support the view that hK6 and hK10 may be involved in insulin and other pancreatic hormone processing and/or secretion, as well as in physiological functions related to the endocrine pancreas. PMID- 12769264 TI - The beta1 integrin subunit is not a specific component of the costamere domain in human myocardial cells. AB - Studies on altered integrin receptor expression during cardiac hypertrophy and heart failure requires accurate knowledge of the distributional pattern of integrins in myocardial cells. At present the general consensus is that in cardiac muscle the beta1 integrin receptor is mainly localized to the same sarcolemmal domain as vinculin at Z-band levels ('costamere'). Since most previous studies have been focusing on myocardial integrin distribution in lower mammals, the myocardial localization of the beta1 integrin subunit was investigated in biopsies collected from the auricle of patients undergoing a coronary bypass operation. Non-invasive serial optical sectioning was carried out by immuno-laser scanning confocal microscopy. Double-labelling for vinculin/alpha actinin, and the cytoplasmic domain for the beta1 integrin subunit, showed that beta1 integrin is deposited throughout both the vinculin/alpha-actinin domains and the non-vinculin/alpha-actinin domains. These results were supported by a semi-quantitative analysis in extended focus images of the latter preparations. Higher magnification views at the electron microscopical levels of the large, extracellular domain of the beta1 integrin subunit disclosed a pronounced labelling in the form of a dense, irregular punctuate pattern that was distributed at Z-disc domains as well as along the entire sarcolemmal area between Z-discs. Our findings show that in human, myocardial cells, the beta1 integrin receptor does not only localize to the surface membrane at the Z-disc level ('costamere' in cardiac muscle), but has a widespread distribution along the sarcolemma. PMID- 12769266 TI - Heparin-binding EGF-like growth factor is seen on the extracellular surface of uterine epithelial cells only after the initial stages of blastocyst attachment. AB - The presence and distribution of heparin-binding epidermal growth factor in rat uterine epithelial cells was determined immunohistochemically and localized ultrastructurally. Rat uterine tissue was examined on days 1, 3, 6 and 8 of pregnancy and it was found that while presence of this growth factor was evident from day 1, spatial reorganization occurred by the time of blastocyst implantation. Strong apical staining was evident from day 6 to day 8, day 6 being the approximate time of blastocyst implantation. Electron microscopy further revealed that this growth factor while shown to be expressed very strongly apically from day 6, actually localized on the plasma membrane only after attachment of the blastocyst. This suggests that heparin-binding epidermal growth factor is not involved in the initial stages of implantation but is more likely involved in the post attachment stages of pregnancy. PMID- 12769265 TI - Expression of aquaporin 3 and its localization in normal skeletal myofibres. AB - The question whether aquaporin 3 (AQP3) is expressed in normal human skeletal muscle at mRNA and protein levels has been examined, since AQP3 has been reported to be coexpressed with AQP4 in various kinds of tissues other than skeletal muscle. The gel electrophoresis of the reverse transcription polymerase chain reaction (RT-PCR) product of total RNA samples extracted from normal human muscle specimens by using the oligonucleotide primers for AQP3 contained a band of 629 base pairs which corresponded to the base pair length between two primers of AQP3. The nucleotide sequence of this RT-PCR product coincided with that of AQP3. At the protein level, immunoblot, immunohistochemical and immunoelectron microscopical studies were done by using rabbit antibody against the synthetic peptide of the cytoplasmic domain of the human AQP3 molecule. Immunoblot analysis showed that rabbit antibody against the human AQP3 reacted with a protein of approximately 30 kDa molecular weight in extracts of normal human skeletal muscles. The immunoreaction for the anti-AQP3 antibody with normal human muscle was noted at the myofibre surface. Immunogold labelling electron microscopy revealed that the gold particles indicating the presence of AQP3 molecules were located mainly at the inside surface of muscle plasma membrane. PMID- 12769267 TI - The nitric oxide synthase-1 and nitric oxide synthase-3/nitric oxide signalling systems in the heart of wild type mice and mouse mutants. AB - Recently, we have shown that nitric oxide synthase-1 (NOS-1) and thus its product NO are present in the sarcolemma region of a subpopulation of atrial cardiomyocytes in the rat heart. In order to find out whether this newly discovered sarcolemma-associated NOS/NO system represents a general signalling mechanism in the murine rodent heart and whether its properties are comparable to those in skeletal muscle fibres, immunohistochemical and catalytic histochemical methods (including image analysis) were applied to the heart and extensor digitorum longus (EDL) and tongue muscles of wild type and mutant mice. In different strains of wild type mice and NOS-3 knockouts, urea-resistant (and therefore specific) NOS NADPH diaphorase histochemistry and NOS-1 immunohistochemistry revealed that NOS-1 activity and protein were present in the sarcolemma region of a subpopulation of atrial and ventricular working cardiomyocytes, but not in those of the impulse conducting system. Using image analysis, NOS-1 showed similar activities in the sarcolemma region of cardiomyocytes and in EDL type I myofibres. In mdx and NOS-1 knockout mice, NOS-1 was absent from the sarcolemma region of atrial and ventricular cardiomyocytes and of EDL and tongue muscle fibres, whereas NOS-1 was present in the hearts of NOS-3 knockouts. Atrial natriuretic peptide immunohistochemistry identified part of the atrial NOS-1-expressing cardiomyocytes as myoendocrine cells. In mdx mice as well as in NOS-1 - and NOS-3-deficient animals, the peptide was found in greater abundance than in wild type mice. These data suggest that NOS-1 is expressed in a subpopulation of working cardiomyocytes in the murine rodent heart, that the myoendocrine cells may be negatively modulated by NOS-1 - and NOS 3-produced NO, and that the anchoring mechanisms for NOS-1 in these cells (i.e. their confinement to the sarcolemma region) are comparable to those in skeletal muscle fibres. PMID- 12769268 TI - In-situ analysis of cellular poly(ADP-ribose) production in scrapie-infected mouse neuroblastoma cells. AB - Transmissible spongiform encephalopathies (TSEs), also called prion diseases, are characterized by formation of the disease-associated isoform of prion protein (PrP(Sc)), which arises from a normal isoform termed PrP(c) by a post translational conversion process occurring in an autocatalytic fashion. Oxidative stress has been proposed as a pathogenetic mechanism in TSEs and increased lipid peroxidation has recently been described in prion-infected cell cultures, suggesting an intrinsic link between the presence of prions and oxidative stress. We investigated if poly(ADP-ribose) formation can be detected in cultured cells upon prion infection, as this NAD+-consuming and DNA strand break-activated nuclear enzymatic reaction has the potential to cause rapid and lethal NAD+ depletion in cells under severe oxidative stress. Poly(ADP-ribose) production was analysed by immunofluorescence in freshly scrapie-infected Neuro2a-D11 mouse neuroblastoma cells, which had been confirmed by immunocytochemistry to produce PrP(Sc), and in uninfected controls. No spontaneous poly(ADP-ribose) specific signals were observed in infected or in uninfected cells, while both cell types readily reacted to H2O2 treatment with poly(ADP-ribose) synthesis in a dose dependent manner, with no obvious difference in staining intensity at any dose tested. In summary, our data reveal that replication of scrapie agent in neuroblastoma cells can proceed without detectable stimulation of the cellular poly(ADP-ribosyl)ation system. PMID- 12769269 TI - Proper nomenclature of formaldehyde and paraformaldehyde fixatives for histochemistry. PMID- 12769270 TI - Grease Cowboy Fever; or, the making of Johnny T. AB - Through a mix of theory, memoir and performance narrative, this chapter examines the making of drag persona Johnny T. as part of a king movement where the dominant cultural paradigm of gender is reconsidered and remastered. As seen in Grease, Saturday Night Fever and Urban Cowboy, pop culture icon John Travolta's particular blend of 50s greaser, faggy 70s disco, and 80s country masculinities are shown to be prime drag king conditions, particularly for a dyke who came of age during the 70s Travolta fever. While drawing from personal experience as a king, current trends in the king movement, and gender theory, this essay calls into question the lines between performing masculinity on and off the stage, inviting us to see both the work and play, the parody and realness, the struggle and liberation that make up the transgressive world of drag kinging and gender variance. Drawing upon gender theorists Judith Butler and Judith Halberstam, gender is exposed as a social construction both produced and performed, and as such, drag kinging is framed as an arena where gender is reconfigured. PMID- 12769271 TI - Me boy. AB - Neeve provides an overview of hir personal and political motivations for drag performance as Pat Riarch. Initially based in a discussion of hir youth's gender game, s/he proceeds to elucidate hir location of adult playmates in the gender bending/drag king community. PMID- 12769272 TI - "I Am the Man!" Performing gender and other incongruities. AB - The following essay details the genesis of the performance "I Am the Man" for the First International Drag King Extravaganza. In it the authors explain both their autobiographical performances and the relationship between their drag performances and transgender theory by linking Shakespeare to queer theory and by overlapping their personal narratives. PMID- 12769274 TI - Lesbian drag kings and the feminine embodiment of the masculine. AB - Part of an ongoing ethnography of an imperial sovereign court I am undertaking, this chapter explores the world of the lesbian drag king and the gendered performance she undertakes in this realm. Taking a relational, situational approach to understanding gender, the lesbian drag queen of the court is also examined in terms of how "her" image and actions give gendered meaning and confer import to the lesbian drag king. Note is also made of lesbian court members' often contradictory gendered relationships with the gay men in this setting: gay drag kings and gay drag kings. Although embodying a masculine persona in image and action has enabled some lesbian drag kings to successfully challenge the often sexist actions and reign of the gay men of the court, it has also resulted in some lesbian drag queens being subordinated in the process. Thus, as much as lesbian drag kings subvert existing gender hierarchies they also sometimes recreate them in the pursuit of situational power. PMID- 12769273 TI - Making Kings. AB - Due to a general lack of representation of female masculinities within the North American media, most urban drag king cultures have evolved in isolation from each other. As a result, drag kings tend to develop locally specific codes of dress, performance styles and forms of masculinity. This chapter describes the case of a group of Montreal drag king performers, The Mambo Drag Kings (MDKs). It is based on interviews that were conducted for Colleen Ayoup's film Kings (2001), a short documentary that explores kinging by examining the development of this specific group of performers. We present excerpts from our discussions regarding the relationships between everyday gender identities, lesbian sexuality, and performing as a king in a society that is fundamentally based on an asymmetrical sex-gender system. While the chapter does highlight these theoretical concerns, the focus is on how the participants of the group interpret and experience kinging and its impact on their own gendered and sexual identities. PMID- 12769275 TI - Capital Drag: kinging in Washington, DC. AB - Through individual interviews with three DC drag kings and detailed, first-person accounts of their performances, I examine the role the kings play within the lesbian community at Club Chaos in Dupont Circle. My interviews address how and why the kings started performing, how their drag characters relate to their everyday personalities and experiences as lesbian women, why performing in drag is important to them, why drag performances are important to the women who attend their shows, and how gay men and drag queens have responded to their performances. My descriptions of the kings' performances, the audience response, and the atmosphere they create at the club reflect my viewpoint as a lesbian audience participant who has much appreciation for drag queens and much curiosity about the burgeoning drag king scene. I conclude that drag kings provide a valuable service to lesbian communities by creating a safe, supportive environment in which lesbian performers and audience members can celebrate and explore their relationships to female masculinities and queer sexualities. PMID- 12769276 TI - Kinging in the heartland; or, the power of marginality. AB - Theorizations of the drag king phenomenon and definitions of "the Drag King" thus far have relied almost exclusively on Judith Halberstam's description of the drag king scenes in New York, London, and San Francisco. In order to expand the scope of the investigation of drag king culture and as an example of the range of drag kings and drag king acts that have developed across the U.S. and across the world, this article focuses on H.I.S. Kings, a group of women, who have been performing in Columbus, Ohio since 1996. By infusing drag with a dose of theory, which is always tempered by their outrageous sense of humor, H.I.S. Kings have developed what arguably is one of the most interesting varieties of kinging, an approach to the genre of the drag show that Halberstam's model cannot account for. Unlike kings in New York, for example, H.I.S. Kings frequently utilize the supposedly gay male form of camp in their performance of multiple masculinities and femininities, and they create innovative ensemble numbers that engage the racial and gender politics of drag. The example of H.I.S. Kings calls into question any theorization of the drag king phenomenon that ignores cultural developments in the heartland by focusing exclusively on cities traditionally considered the centers of queer culture. PMID- 12769277 TI - Walk like a man: enactments and embodiments of masculinity and the potential for multiple genders. AB - With this chapter, I explore the potential of drag king performance as a tool of deconstructing gender. I begin with a brief examination of the ways that gender is constructed as natural through the repetition of a set of norms, noting the pervasiveness of these norms and their location at the center of cultural imagination. I then turn to the idea of drag as a practice of subverting these norms through breaking their repetition. I argue that drag king performance, through its failure to approximate the "natural male," draws attention to the very constructedness of the category, and thus of all naturalized categories of gender. Using the budding Edmonton drag king scene as a case example, I discuss some of the ways that drag kings take queer theory and gender deconstruction out of the classroom and into practice. I speculate as to the effectiveness of such practices in the realm of queer theory, pointing to the ways that these performances may be read to reinforce rather than subvert gender norms. I also look at the complexities of "passing" gender performances, such as transsexual or transgendered practices, and the ways that these practices align or do not align with performances intending to fail. I propose that the emergence of these practices is in itself a beginning step toward a reevaluation of normative gender, and toward a radical reinvention of new gender possibilities. PMID- 12769278 TI - Drag kings in the new wave: gender performance and participation. AB - In an examination of Midwestern drag king performers and communities that have emerged since the study by Volcano and Halberstam of king cultures in London, New York, and San Francisco, this article considers traditional and alternative ways of "doing drag," both performative and participatory, as a means of interrogating the proximity of a "new wave" of king culture to academic theory. Tracing the evolution of drag king performance in the Twin Cities from the 1996 workshop by Diane Torr to the formation of two distinct king troupes in the late 1990s demonstrates a particular trajectory in kinging that reflects a new consciousness and enactment of gender theory through artistic praxis. Participation plays a key role in breaking down the distance between spectator and performer in venues such as the First International Drag King Extravaganza in Columbus, Ohio, and Melinda Hubman's art installation "Performing Masculinities: Take a Chance on Gender" in Minneapolis. By engaging the "audience" in drag, the Extravaganza "Science Fair" successfully referenced drag kings' shared history with early American freak shows in a clever and critical way. Moving beyond the contest framework of early king shows, new drag king troupes like Minneapolis' Dykes Do Drag are "mixing it up" in an attempt to complicate notions of butch/femme gender roles, sexuality, and drag stereotypes. PMID- 12769279 TI - A voice from the audience. PMID- 12769280 TI - One body, some genders: drag performances and technologies. AB - This chapter examines the ways in which one body becomes the site for multiple and varied gender performances through Alana Kumbier's performances as drag king Red Rider and drag queen Red Pearl. Using theoretical frameworks provided by Judith Halberstam, Ira Livingston, and Teresa de Lauretis the essay also calls for a consideration of material technologies and artifacts as technologies of gender. PMID- 12769281 TI - Drag king magic: performing/becoming the other. AB - This chapter seeks to theorize drag king practice through the lenses of alterity, liminality, and performance theory, while attempting to complicate and reinvigorate discussions of identity raised by drag. I examine the ways in which drag king performance plumbs the concept of "the Other," and forces confrontation with a complex field of desire. Contemporary "queergirl" existence negotiates a range of desirable and desiring Others, from the polarities (i.e., butch-femme) unique to queer structures of desire, to the desire of those on the cultural margins for the power of those at the center, and vice versa. I employ anthropological theories of performance, mimesis, and liminality to establish a framework through which drag kings may be viewed as crucibles of this desire and agents of this power exchange. By performing maleness, drag kings expand and redraw the definitional boundaries of the male, interfere with the cultural power of mainstream maleness, and simultaneously transfer some of this power to themselves as queer women. At the same time, drag king existence forces a renegotiation of queergirl desire to encompass a range of masculinities. By performing/becoming the Other, drag kings engage in a practice of magic which transforms both margin and center. PMID- 12769282 TI - Erotic arguments and persuasive acts: discourses of desire and the rhetoric of female-to-male drag. AB - This chapter analyzes the October 21, 2000 drag king performance by Mildred Gerestant, a.k.a. Dred. Arguing that her act appropriates, embodies, and manipulates certain ideological discourses of desire and identity, the article studies the shape and force of Dred's performative arguments and illuminates how a drag king act can not only tease and titillate the addressed audience (the actual people who watched the show) but also hail the invoked audience (the audience called upon, imagined, or made possible by the performance). Arguing that, as a rhetorical act, Dred's performance offers a purposeful discourse about gender, race, and power, this chapter ultimately explores how a rhetorical analysis of FTM drag elucidates the complex relationship between the rhetor, performance, and audience. More specifically, it shows how a rhetor's performance of non-normative identity and engagement with discourses of desire has the potential to unmask the hegemonic effects of language and power and to unsuture the seemingly natural connections between sex and gender. PMID- 12769283 TI - Seeing double, thinking twice: the Toronto drag kings and (re-) articulations of masculinity. AB - Through a close reading of the performances of masculinity by the Toronto drag kings, this chapter argues that drag king shows parody the hyper-masculine star at his most contradictory and dialogic. Given that drag king performances parody both the contradictions of masculinity on stage, and the productive technologies of the star, king performances are essentially both meta-theatrical (performances about performing where lights, music, body language, dance all make the man) and meta-performative (performances which are at once conditioned by the performative reiterations which enable a fiction of identity in the first place). Finally, I explore the rather abstracted question of what cultural work the category of "drag king" does. I argue that it is a term which articulates a series of productive but necessary slippages in and through the contradictory and dialogic practices of identification. The bottom line is this: drag kings are situated in and play with the ironic no man's land between "lesbian," "butch," "transman" and "bio-boy" where the sell evident is neither. PMID- 12769284 TI - Whose drag is it anyway? Drag kings and monarchy in the UK. AB - This chapter will show that the term "drag" in drag queen has a different meaning, history and value to the term "drag" in drag king. By exposing this basic, yet fundamental, difference this paper will expose the problems inherent in the assumption of parity between the two forms of drag. An exposition of how camp has been used to comprehend and theorise drag queens will facilitating an understanding of the parasitic interrelationship between camp and drag queen performances, while a critique of "Towards a Butch-Femme Aesthetic," by Sue Ellen Case, will point out the problematic assumptions made about camp when attributed to a cultural location different to the drag queen. By interrogating the historical, cultural and theoretical similarities and differences between drag kings, butches, drag queens and femmes this paper will expose the flawed assumption that camp can be attributed to all of the above without proviso, and hence expose why drag has a fundamentally different contextual meaning for kings and queens. This chapter will conclude by examining the work of both Judith Halberstam and Biddy Martin and the practical examples of drag king and queen performances provided at the UK drag contest held at The Fridge in Brixton, London on 23 June 1999. PMID- 12769285 TI - Drag kings "down under": an archive and introspective of a few Aussie blokes. AB - The mid 1990s saw an explosion of Drag Kings in many major and smaller cities throughout the world. While documentation of this has largely occurred through publications in the USA and UK, the Internet and smaller publications have demonstrated a phenomenon that has arguably re-ignited feminist debate. In Adelaide, Australia, Ben Dover and His Beautiful Boys set the annual lesbian and gay festival alight. This chapter describes this performance to set the stage for exploration of some of the workings of 'race' and ethnicity in the creation of persona, choice of name and naming that is brought to Drag King performance. Drawing on interview material the chapter suggests that just as Drag Kings and kinging has been a useful and provocative site for closer and deeper understandings of genders, bodies and sexualities, Drag Kings and Kinging may also provide a useful site for unraveling some of the minefield that is race and racism. PMID- 12769286 TI - Cytogenetic tools for Arabidopsis thaliana. AB - Although the first description of chromosomes of Arabidopsis dates as far back as 1907, little attention was paid to its cytogenetics for a long time. The spectacular interest in chromosome research for this species that now is the model plant species by excellence came with the introduction of molecular cytogenetical research including FISH technology, genome sequence data and immunodetection of chromatin proteins. In this paper, we present an overview of the most important cytogenetic tools that were developed for Arabidopsis in recent decades. It shows the power of meiosis for studying synaptic mutants and FISH technology, and the development of numerical and structural chromosome mutant series like trisomics, telotrisomics and translocations for assigning linkage groups to chromosomes. Its small genome and chromosome size and relatively simple organization of heterochromatin have been the key to a successful characterization of the molecular organization of repetitive and single copy sequences on the chromosomes, both in metaphase and pachytene complements, but also in interphase nuclei and extended DNA fibres. Finally, Arabidopsis is the first plant species in which a heterochromatin knob could be analysed in full detail and in which chromosome painting with BAC clones covering whole chromosome arms could be established. All these achievements are probably only the very first steps in a promising new era in plant cytogenetics and chromatin research yet to come. PMID- 12769287 TI - Recent progress in chromosome painting of Arabidopsis and related species. AB - This paper reports on state-of-the-art achievements of chromosome painting in Arabidopsis thaliana (2n = 10). Arabidopsis chromosomes 1, 2 and 4 were painted using chromosome-specific BAC contigs. We consider technical aspects of the painting approach and document major applications, such as the tracing of Arabidopsis chromosomes as interphase chromosome territories and during mitotic and meiotic cell cycles as well as comparative chromosome painting in related species. This is the first report of successful interspecific chromosome painting in plants. The evolutionary history of chromosomes homeologous to Arabidopsis chromosome 4 was reconstructed by hybridization of chromosome-4-specific painting probes to karyotypes of Brassicaceae species with x = 8 chromosomes. Future perspectives of chromosome painting in A. thaliana and its wild relatives are outlined. PMID- 12769288 TI - Meiotic chromosome synapsis and recombination in Arabidopsis thaliana; an integration of cytological and molecular approaches. AB - Arabidopsis has emerged as an important model for the analysis of meiosis in Angiosperm plants, creating an interesting and useful parallel to other model organisms. This development has been underpinned by advances in the molecular biology and genetics of Arabidopsis, especially the determination of its entire genome sequence. However, these advances alone would have been insufficient without the development of improved methods for cytological analysis and cytogenetic investigation of meiotic nuclei and chromosomes. A basic descriptive framework of meiosis in Arabidopsis has been established based on these procedures. In addition, molecular cytogenetic and immunocytological techniques have provided supplementary detailed information on some aspects of meiosis. Gene identification and characterization have proceeded in parallel with these developments based on both forward and reverse genetic procedures utilising the considerable range of Arabidopsis genetic and molecular resources, such as T-DNA and transposon tagged lines as well as the genomic DNA database, in combination with cytological analysis. A diverse range of meiotic genes have been identified and analysed by these procedures and in selected cases they have been subjected to detailed functional analysis. This review focuses on genes that are involved in the key meiotic events of chromosome synapsis and recombination. PMID- 12769289 TI - FISH analysis of meiosis in Arabidopsis allopolyploids. AB - Although allopolyploids are common in nature and in agriculture, knowledge of their origin, evolution and genomic regulation is limited. We study synthetic allotetraploids of Arabidopsis thaliana and Arabidopsis arenosa as well as the natural allotetraploid Arabidopsis suecica. To elucidate the composition and behavior of the allotetraploid genome, we used chromosome painting with probes from contiguous regions of chromosome 4 of A. thaliana and fluorescent in-situ hybridization with centromeric (CEN) probes specific for each parental genome. We documented the presence of 16 A. arenosa and 10 A. thaliana chromosomes and demonstrate that two different A. arenosa chromosomes are homeologous to chromosome 4 of A. thaliana. Although chromosome pairing in pollen mother cells was predominantly homologous, CENs of different parental origin coalesced at early prophase I, but resolved into proper pairs by metaphase. In addition, CENs of homologous chromosomes were not paired in tapetum cells and endopolyploidy without strict polyteny was evident by the large number of independent CENs. Thus, the Arabidopsis synthetic allopolyploids were capable of homologous pairing as early as three generations after their formation. This indicates that diploid like pairing is not the result of adaptive mutations in genes that regulate pairing nor the result of structural remodeling of the genomes: rather, it is likely that either the parents provided genes controlling pairing behavior or that features of the parental chromosomes hinder homeologous pairing. PMID- 12769290 TI - Heterochromatin in interphase nuclei of Arabidopsis thaliana. AB - The eukaryotic nucleus represents a complex arrangement of heterochromatic and euchromatic domains, each with their specific nuclear functions. Somatic cells of a multicellular organism are genetically identical, yet they may differ completely in nuclear organization and gene expression patterns. Stable changes in gene expression without modifying the sequence are the result of epigenetic changes and include covalent modifications in cytosine residues of DNA and in histone tails giving rise to altered chromatin protein complexes, remodeling of chromatin and changes in chromatin compaction. Large-scale differences in chromatin structure are visible at the microscopic level as euchromatin and heterochromatin. Arabidopsis thaliana chromosomes display a relatively simple distribution of euchromatic and heterochromatic segments overlapping with gene rich and repeat-rich regions, respectively. Recently, we have shown that Arabidopsis provides a well-defined system to study individual chromosomes and chromatin domains in interphase nuclei as well as the relationship between chromatin condensation and epigenetic mechanisms of gene silencing. This overview focuses on the organization and composition of heterochromatin in Arabidopsis nuclei. PMID- 12769291 TI - Tandemly repeated DNA sequences and centromeric chromosomal regions of Arabidopsis species. AB - Despite their common function, centromeric DNA sequences are not conserved between organisms. Most centromeres of animals and plants so far investigated have now been shown to consist of large blocks of tandemly repeated satellite sequences that are embedded in recombination-deficient heterochromatic regions. This central domain of satellite sequences that is postulated to mediate spindle attachment is surrounded by pericentromeric sequences incorporating various classes of repetitive sequences often including retroelements. The centromeric satellite DNA sequences are amongst the most rapidly evolving sequences and pose some fundamental problems of maintaining function. In this overview, we will discuss work on centromeric repetitive sequences in Arabidopsis thaliana and its relatives, and highlight some of the common features that are emerging when analysing closely related species. PMID- 12769292 TI - Using Arabidopsis to understand centromere function: progress and prospects. AB - Arabidopsis thaliana has emerged in recent years as a leading model for understanding the structure and function of higher eukaryotic centromeres. Arabidopsis centromeres, like those of virtually all higher eukaryotes, encompass large DNA domains consisting of a complex combination of unique, dispersed middle repetitive and highly repetitive DNA. For this reason, they have required creative analysis using molecular, genetic, cytological and genomic techniques. This synergy of approaches, reinforced by rapid progress in understanding how proteins interact with the centromere DNA to form a complete functional unit, has made Arabidopsis one the best understood centromere systems. Yet major problems remain to be solved: gaining a complete structural definition of the centromere has been surprisingly difficult, and developing synthetic mini-chromosomes in plants has been even more challenging. PMID- 12769295 TI - Changes in intestinal electrical activity during ischaemia correlate to pathology. AB - The gastrointestinal tract possesses an omnipresent electrical slow wave called the basic electrical rhythm (BER). It has been shown that the frequency of the BER falls during intestinal ischaemia. The correlation between changes in the BER and pathology that occur during acute ischaemia are not completely understood. To study this, the electrical activity of the ileum in 14 adult male rabbits was recorded during ischaemia. At baseline, 60, 120 and 210 minutes of ischaemia, segments of bowel were resected for histopathologic evaluation. The BER frequency was determined using the Fast Fourier Transformation (FFT) spectral analysis. The BER amplitude and FFT spectral power were also determined. The results showed significant decrease (p<0.05, Student's T-test) in the BER frequency, amplitude, and spectral power at all time points. Between 60 and 120 minutes, while there was a decrease in BER activity the pathologic grade remained the same (focal loss of surface epithelium). By 210 minutes of ischaemia when the BER could not be recorded, there was diffuse mucosal infarction. The results indicate that changes in the electrical activity of the bowel during acute mesenteric ischaemia occurred prior to the pathologic changes. The presence of electrical activity indicates that there was viable bowel. Thus it should be possible to use recordings of electrical activity to evaluate bowel viability during acute ischaemia. PMID- 12769296 TI - Perceived, desired, and normatively determined orthodontic treatment needs among orthodontically untreated Nigerian adolescents. AB - This study assessed perceived, desired, and normative need for orthondontic care in a randomly selected (n=567) Nigerian children aged 12-18 years (mean age, 14.6 +/- 1.5), in Ibadan city. Perceived and desired needs were collected using a pre tested questionnaire. Normative need was assessed on all participants by one orthodontist using the Dental Aesthetic Index. Results revealed 13.8% of the children having very severe or handicapping malocclusion with treatment considred mandatory; 9.7% with severe malocclusion and treatment highly desirable; 19.0% having definite malocclusions with treatment elective and 57.5% had normal or minor malocclusions with no treatment or slight treatment need. About 48.4% of the children desired orthodontic care and 81.7% perceived the need for orthodontic care. No statistically significant gender differences (P>0.05) were observed in perceived, desired and normatively determined orthodontic treatment needs as well as between socioeconomic backgrounds. Although considerable proportion of the adolescents perceived, desired and objectively needed orthodontic care, a discrepancy was observed as some who had near 'ideal occlusion' felt the need for treatment while some who had handicapping malocclusion felt otherwise. Therefore, in orthodontic counselling of Nigeria adolescents, attention should be paid to how the child perceives his/her dentition. PMID- 12769293 TI - Telomere structure, function and maintenance in Arabidopsis. AB - The stability of eukaryotic genomes is provided in part by the integrity of telomeres, the nucleoprotein caps on the ends of chromosome. Recent studies reveal that proper telomere architecture is required for long-term proliferation capacity. Here we describe molecular mechanisms that protect and maintain chromosome ends and discuss why Arabidopsis is emerging as a powerful new model for elucidating fundamental aspects of telomere biology. PMID- 12769297 TI - Cost effectiveness of autologous blood transfusion--a developing country hospital's perspective. AB - An autologous blood donation program was set up at National Orthopaedic Hospital, Igbobi Lagos in 1992 in response to the rising sero prevalence of HIV observed in our "relative replacement" donors. A retrospective batch analysis of patients who received autologous transfusion and those who received homologous blood in our hospital in 1997 was carried out. Based on hospital charges, the mean charge (from the day of operation and excluding the cost of surgery) was dollars 116 (+/ dollars 7), median dollars 102 for those who donated and used their own blood compared to the mean charge of dollars 259.7 (+/- 116.3), median dollars 224, for homologous blood recipients (P=008). This was found to be due to a significant difference in the means of length of hospital stay of 21 days for autologous blood recipients, 34 for homologous blood recipients (P=0.009). The rate of infection was 85.7% for homologous blood recipients and 14.3% for autologous blood recipients. There was no significant difference in the means hospital charges, length of hospital stay and rate of infection in the entire population of patients who received blood transfusion when analysed by ward and consultant. We conclude that homologous blood transfusion in this hospital is significantly more expensive than autologous transfusion mainly due to greater infective morbidity in homologous blood recipients. PMID- 12769298 TI - Treatment of ganglion using hypertonic saline as sclerosant. AB - Twenty-nine patients with ganglion of the wrist were treated in this hospital using hypertonic saline as sclerosant. All patients were treated as outpatients. Under aseptic conditions, the ganglia were aspirated using #18 hyperdermic needle. A mixture of 2 cc of hypertonic saline and 1 cc 1% xylocaine were injected into the empty cavity and crepe bandage applied for 24-48 hours. After a follow-up period of 24-36 months, there was only one recurrence which was believed to be accidental injection of the saline outside the empty cavity. This was treated by the same procedure. The most common complication was swelling of the wrist and dorsum of the hand which were seen in 50% of cases. This subsided spontaneously within 72 hrs of treatment. Severe pain necessitating ingestion of analgesics (Paracetamol) was reported in 6 patients (20%), which subsided within 48 hours. It is hoped that this new treatment which is cheap and less invasive may be a break through in the treatment of ganglia which hitherto was characterised by high recurrence rate of up to 23%. PMID- 12769299 TI - Prevalence of atopic diseases in Nigerian children with vernal kerato conjunctivitis. AB - A descriptive study of 117 cases of vernal kerato-conjunctivitis (VKC), in children below the age of 18 years, newly presenting to 2 hospitals in Ibadan, Nigeria. All the children had a structured questionnaire administered to them which inquired about demographic and clinical, especially history or presence of other atopic diseases amongst other things. Specifically inquired about were asthma, eczema, allergic rhinitis or hay fever, allergic skin rash e.g. scabies, reaction to drugs and others. The children were also examined to confirm or detect the presence of these atopic diseases. The overall prevalence of atopic conditions was 19.8% amongst cases of VKC. These comprised of 6% asthma, 5% allergic rhinitis and 4.3% eczema. (Total 15.5%). Other forms of atopic conditions like other non-specific allergic skin reactions (0.9%), reaction to certain drugs e.g. chloroguine (0.9%) and other unknown allergic conditions (2.6%). It is suggested that children with VKC should be subjecteed to detailed enquiry and examination with regards to the presence of other atopic conditions. This will go a long way in understanding the type(s) of allergic response involved and perhaps of management of VKC. PMID- 12769300 TI - Maternal haematocrit and pregnancy outcome in Nigerian women. AB - Anaemia is the most common medical disorder in pregnancy and a direct or indirect cause of maternal and perinatal mortality; therefore antenatal care should be concerned with early detection and management. The incidence of anaemia in 735 normal singleton pregnant patients at booking in the University College Hospital, UCH, Ibadan, between the 1st of June 2000 and 31 of May 2001 was 15%. Six hundred and thirty-three of the patients (86.1%) presented for delivery at this centre. Five hundred and sixty seven (89.6%) of the patients were not anaemic when seen in labour, reducing the incidence of anaemia to 10.4%. Forty-two (6.6%) and 24 (3.8%) patients had mild and moderate anaemia respectively. No patient had severe anaemia. Of those presenting in labour, 195 (30.8%) were primigravid, while 417 (65.9%) and 21 (3.3%) were multiparous and grandmultiparous respectively. Five hundred and twenty-eight (83.4%) were aged between 21 and 35 years. The mean gestational age at booking was 21.3 weeks and at delivery was 38.7 weeks. Spontaneous vertex delivery was achieved in 76.3% with 96.7% having live births. The perinatal mortality rate was 33 per 1,000 births. Stillbirths occurred more in the moderately anaemic patients. The higher the packed cell volume in labour, the greater the birth weight, better the Apgar scores but the more the blood loss at delivery. The babies of the patients with a normal packed cell volume had better Apgar scores at one minute, which was statistically significant (p value < 0.05), but the mildly anaemic patients had babies with better Apgar scores at 5 minutes. This was however not statistically significant. There was no maternal death. PMID- 12769301 TI - A review of typhoid perforation in a rural African hospital. AB - In Wesley Guild Hospital Ilesa in the South-West region of Nigeria a retrospective study of 105 consecutive cases operated upon for typhoid perforation between January 1988 and November 2001 was carried out. The ages of the patients were between 4 to 70 years with a mean age of 19.2 +/- 8.81. There were 84 males (80%) and 21 females (20%) giving a ratio of 4:1. Diagnoses were based on clinical and radiological findings. All the patients had laparotomy after resuscitations with intravenous fluids, electrolytes replacement, broad spectrum antibiotics, Nasogastric intubation/suctioning and urethral catherterisation. There were five negative laparotomies (4.8%). Eighty patients (76.2%) had a single perforation, while the rest 20 had multiple perforations. The perforations were located between 7 cm and 100cm from the ileo-Caecal junction. Apart from the patients who had resection and primary anastomosis, 95 (90.5%) had 2 layered closure of the perforation. The most common complications were wound infections (26.7%). Intra-abdominal abscesses (9.5%) and would dehiscence (7.6%). The mortality rate was 16.2% showing a remarkable improvement in the West African Subregions. PMID- 12769302 TI - Reviewing trends in the incidence of cancrum oris in Ibadan, Nigeria. AB - The objective of this clinic based retrospective study was to review the trends in the incidence of cancrum oris at the Dental clinic of the University College Hospital, Ibadan, Nigeria. Records of children aged 1 to 16 years diagnosed as having cancrum oris between 1st of January 1986 to 31st December, 2000 were reviewed. Of the six thousand three hundred and ninety (6390) children seen within the period of study, 45 had cancrum oris with the modal age been 3-5 years and the mean age was 4.2 +/- 2.7 years. There was a declining trend in the proportion of children presenting with the disease at five yearly interval within the period of study, although malnutrition was still a common factor in all the children. PMID- 12769294 TI - Chromatin dynamics and Arabidopsis development. AB - The plant life cycle involves a series of developmental phase transitions. These transitions require the regulation and highly co-ordinated expression of many genes. Epigenetic controls have now been shown to be a key element of this mechanism of regulation. In the model plant Arabidopsis, recent genetic and molecular studies on chromatin have begun to dissect the molecular basis of these epigenetic controls. Chromatin dynamics represent the emerging and exciting field of gene regulation notably involved in plant developmental transitions. By comparing plant and animal systems, new insights into the molecular complexes and mechanisms governing development can be delineated. We are now beginning to identify the components of chromatin complexes and their functions. PMID- 12769303 TI - Aetiological agents, clinical features and outcome of septicaemia in infants in Ibadan. AB - The present study sought to determine the prevalence, clinical features and bacterial aetiological agents of septicaemia in post-neonatal infants (age 1-12 months) presenting with fever at the University College Hospital, Ibadan, Nigeria. STUDY SETTING: It was carried out at the Otunba Tunwase Children Emergency Ward (OTCHEW) (a 40 bedded ward) of the Department of Paediatrics, University College Hospital, Ibadan, Nigeria. PATIENTS: All postneonatal infants aged 1-12 months presenting with fever and who had not had antibiotic treatment in the week prior to presentation during the period June to November 1998 were enrolled in the study. Each child had a full clinical evaluation followed by a blood culture. RESULTS: The infants comprised 56 (54.9%) males and 46 (45.1%) females. The mean age was 5.6 (SD 0.3) months. The prevalence of septicaemia in the infants studied was 38.2%. Clinical features associated with increased risk of septicaemia among these infants were age < or = 6 months, restlessness and a total white cell count > or = 15000/mm3. The organisms isolated in the infants studied were Escherichia coli (35.9% of positive cultures). Staphylococcus aureus (33.3%), Klebsiella species (10.3%), Streptococcus species (7.7%), Proteus species (5.1%), Pseudomonas species (5.1%) and Salmonella species (2.6%). Mortality was significantly higher in patients with septicaemia (25.7% compared with those without septicaemia (7.9%). CONCLUSIONS: Thirty eight percent of febrile infants presenting in this study had positive bacteria blood cultures, the most common organisms being Escherichia coli and Staphylococcus aureus. Few clinical features distinguished febrile infants with septicaemia from those without. It is recommended that febrile infants in our setting with clinical features associated with increased risk of septicaemia should be treated emprically with antibiotics since the probability of having septicaemia is significant. PMID- 12769304 TI - Encephalocele and associated skull defects. AB - Encephalocele is a common congenital problem in the practice of Neurosurgery worldwide, with varying sizes of the underlying skull defects. This study was carried out to determine the size of the problem; to assess whether the skull defects are being under-managed or not; and also to determine those patients that will benefit from cranioplasty. The case notes of the patients with encephalocele managed over a 5 year period were reviewed and the relevant data obtained. Seventy-six percent of the patients had occipital encephalocele. The average diameter of the skull defect was 1.8 cm. Only 2 (9.5%) of the patients had cranioplasty. Cosmesis was acceptable to all the patients. No recurrence was noted in the series studied. We therefore concluded that the skull defects are not being under-managed, however large anteriorly based lesions with wide skull defects (i.e >2.5 cm) will require cranioplasty. PMID- 12769305 TI - Pregnancy outcome in singleton term breeches from a referral hospital in Saudi Arabia. AB - OBJECTIVE: To compare the pregnancy outcome in women with singleton breech presentation at term delivered by caesarean section (CS) and vaginal breech delivery. DESIGN: A retrospective study. SETTING: Abha Maternity hospital, Saudi Arabia. MATERIAL AND METHODS: 573 women with singleton breech presentation at term who delivered between January 1994 and December 2000 formed the basis of this study. There were 166 patients (28.9%) who had assisted vaginal breech delivery (AVBD) and 407 patients (71.1%) who were delivered by CS. RESULTS: There were no statistically significant differences in the mean age and number of abortions between the two groups, (p>0.05) but statistically significant differences were found regarding the birth weight and parity respectively (p<0.05). One hundred and fourteen (19.7%) of patients had a previous CS, and of these 2 (1.75%) delivered vaginally. Caesarean section was carried out electively in 161 (39.5%) of the 407 who had caesarean delivery. There were no statistically significant differences in the perinatal mortality rates, congenital malformation rates and Apgar score of less than 7 at 5 minutes in babies born by AVBD and CS, (p>0.05). There was statistically significant difference in birth trauma (p<0.00001). CONCLUSION: Vaginal breech delivery is strongly associated with birth trauma in our community. It is recommended that attention should be given to trainee obstetrician in selective external cephalic version at term and also the procedure of AVBD so as to reduce the caesarean section rate and also neonatal morbidity in term breeches in our community. PMID- 12769306 TI - Childhood heart failure in Ibadan. AB - One hundred consecutive admissions of children with a diagnosis of congestive cardiac failure to the paediatric department of the University College Hospital, Ibadan were evaluated during a 10-month period. They were aged 8 days to 12 years. They constituted 5.8% of the total paediatric admissions during the study period. The most frequent underlying causes of heart failure were acute lower respiratory tract infections (36%), intrinsic heart disease (31%) and severe anaemia (28%). Other less common causes of heart failure were renal disorders (3%) and septiceamia (2%). Heart failure remains a common problem encountered in emergency paediatric practice in Nigeria, with the majority of cases resulting from preventable causes. PMID- 12769307 TI - Mandibular segmental reconstruction with iliac crest. AB - Twenty patients consisting of 14 males and 6 females with benign destructive lesions of the mandible were reconstructed using free nonvascularised iliac crest. Harvested bone was contoured and secured with 0.5 mm stainless steel wire and reinforced with maxillo-mandibular fixation. Five patients has hemimandibulectomy with immediate reconstruction. The other 15 patients had 1 to 3 segments of the mandible reconstructed. There was only one failure. Mouth opening and closure were centric except in the patients that had hemimandibulectomy without condylar reconstruction. Mastication and facial appearance were satisfactory. In conclusion, the iliac crest is recommended for reconstruction of hemimandible as well as long contiguous segments of the mandible. PMID- 12769308 TI - A survey of common pathogens in wound in patients at the University of Port Harcourt Teaching Hospital (U.P.T.H), Port Harcourt. AB - During a 30-month period, January 1996 to June 1998, a total of 2458 wound swab samples were collected, cultured and identified. In descending order of frequency, the organisms isolated include: Staphylococcus aureus (31.60%), Escherichia coli (25.97%), Pseudomonas aeruginosa (21.21%), Klebsiella species (10.82%), Proteus species (8.23%), non-haemolytic streptococci (1.29%), Beta haemolytic streptococcus (0.43%) and Candida albicans (0.43%). The variation in frequency of the isolates according to sex, age and ward is reported. The antibiotic sensitivity pattern of these isolates is also presented. PMID- 12769309 TI - Factors affecting the survival of the "at risk" newborn at Korle Bu Teaching Hospital, Accra, Ghana. AB - High risk pregnancies continue to be associated with high perinatal mortality and morbidity in developing countries. Korle Bu Teaching Hospital is no exception with a perinatal mortality rate of 98.7/1000 births. Multiple factors resulting in this include the high risk nature of the pregnancies resulting in increased incidence of premature deliveries and asphyxiated babies, the delay in transfer of the sick neonate as well as the inadequate mode of transfer. The type of delivery other than the spontaneous vaginal route also affects the outcome, though the relationship was not statistically significant. Logistic regression analysis showed that maturity, birthweight and time from birth to admission to NICU were the most significant factors associated with the survival of the neonate. Proper foetal surveillance both in the antenatal period and during labour cannot be over emphasized and the mere presence of a paediatrician at these high risk deliveries may make a difference. Also, increased vigilance in the special care offered will help reduce mortality. PMID- 12769310 TI - Histopathologic changes during mesenteric ischaemia and reperfusion. AB - The basic electrical rhythm (BER) of the intestine is known to decrease during mesenteric ischaemia. Some studies have reported the relationship between the BER and the pathologic changes that occur in the bowel during vascular injury. However, these changes have not been completely elucidated. This study describes the histopathologic pattern when the rabbit small intestine was subjected to ischaemia of varying time lengths (30-150 minutes) and subsequent reperfusion for six hours. Intestinal biopsies were taken at baseline, at the end of ischaemia, and at hourly intervals during reperfusion. Microscopic examination of the biopsies revealed evidence of progressive infarction of the mucosa during ischaemia. There was an acute worsening of the pathology during reperfusion, the severity being greater when reperfusion was preceded by longer periods of ischaemia. These changes were statistically significant. The observed pattern in this study shows clearly that reperfusion injury is reflected in the histopathologic response and that this is worse in severity than the response to ischaemia. Studies of longer duration should further clarify the picture during recovery in ischaemia/reperfusion injuries of the bowel. PMID- 12769311 TI - Comparing permanent tooth sizes of Nigerians and American Negroes. AB - A study of two hundred and fifty secondary school children (125 males and 125 females) in Ibadan metropolis was done, to determine mean tooth sizes for the population. The data obtained in Nigerians were then compared with a similar data obtained in American Negros by Richardson and Malhotra (1975). This was done on computer using the Student T-test and taking a probability value greater than 1% (P>0.01) as not significant. The results showed that there was no significant difference in tooth sizes of Nigerians and American Negroes. This could be genetically linked, signifying that Black Americans may have originated from Africa. PMID- 12769312 TI - Domiciliary treatment of femoral shaft fracture in children. AB - A total of 20 children presenting in Accident and Emergency (A&E) Department of University College Hospital, Ibadan, Nigeria with femoral shaft fracture treated with straight leg fixed traction in a domiciliary setting is presented. The male:female ratio is 3.2 with fall accounting for 65% of the cases. There was satisfactory correction of overriding and angular deformity following the application of the fixed traction in all the patients. There was good Callus formation at six weeks with no vascular, neurological or soft tissue complication. PMID- 12769313 TI - Nosocomial significant bacteriuria: prevalence and pattern of bacterial pathogens among children hospitalised for non-infective urinary tract disorders. AB - This study was undertaken to determine the prevalence of nosocomial significant bacteriuria (NSB) and pattern of bacterial pathogens among children hospitalised for non-infective urinary rtract disorders (NUTDs) namely, acute glomerulonephritis, nephrotic syndrome, renal failure and congenital urinary tract anormalies in our nephrology unit. Serial midstream or suprapubic puncture urine specimens were collected into sterile plastic universal bottles from the patients for microscopy, culture and sensitivity, using the standard laboratory methods. Seventeen of the 96 patients admitted for NUTDs were excluded from the study based on the set exclusion criteria; only 19 out of the 79 patients studied were found to have NSB giving a prevalence rate of 24.05%. The isolated pathogens included Klebsiella spp. (47.37%). Staphylococcus aureus (31.58%). Escherichia coli (10.53%), Pseudomonas spp. (5.26%) and Citobacter spp. (5.26%). While 78.95% and 69.23% of the isolates were sensitive to gentamicin and cefuroxime respectively, 57.9% were sensitive to ceftazidime and nitrofurantoin. Less than 50% of the isolates were sensitive to each of the other antibiotics tested. Five of the patients died giving a case fatality rate of 26.3%. In this study, NSB is evidently a very common health problem and a significant risk factor for mortality in patients with NUTDs. Weekly urine culture is recommended in any hospitalised child with NUTD in order to diagnose and manage NSB early before clinical deterioration sets in. PMID- 12769314 TI - Surgical audit in the developing countries. AB - Audit assures provision of good quality health service at affordable cost. To be complete therefore, surgical practice in the young developing countries, as elsewhere, must incorporate auditing. Peculiarities of the developing countries and insufficient understanding of auditing may be, however, responsible for its been little practised. This article, therefore, reviews the objectives, the commonly evaluated aspects, and the method of audit, and includes a simple model of audit cycle. It is hoped that it will kindle the idea of regular practice of quality assurance by surgeons working in the young developing nations and engender a sustainable interest. PMID- 12769315 TI - Residency training in the United States: what foreign medical graduates should know. AB - This communication provides useful information for Foreign Medical Graduates (FMGs) planning to pursue post-graduate residency training in the United States of America (USA). While the number of residency training positions is shrinking, and the number of United States graduates has steadily declined over the past decade, the number of FMGs has also steadily increased. Unfortunately, as a result of the terrorist attack of September 11, 2001, the problem of obtaining training visas and sponsorship by the Educational Commission for Foreign Medical Graduates (ECFMG) will remain major problems for FMGs. This present review is by no means exhaustive. It should serve as a guideline to more detailed and focused information. It is extremely important to network with friends, family, and professional colleagues regarding the subjective and objective aspects of pursuing graduate medical education (GME) in the USA. Though the process is very protracted and at times frustrating and depressing; the visa hurdle will be the ultimate barrier to overcome. However, majority of FMGs eventually achieve the ultimate goal of obtaining superb training and experience in the United States. We highly recommend the book by Kenneth V. Iserson entitled "Iserson's Getting Into A Residency: A Guide for Medical Students" published by Galen Press Limited, Tucson, Arizona (ref# 10). It is a very useful source of information for medical students and residents planning further residency training in the United States of America. Useful Internet addresses are listed in Appendix 1. PMID- 12769316 TI - Sudden infant death syndrome (SIDS) or cot death: a review. AB - Sudden death of an infant is not an uncommon event and when there is no adequate cause to explain the death it poses a diagnostic dilemma for the clinician, and makes discussion of the cause of death with parents quite difficult and unsatisfactory. Sudden infant death syndrome (SIDS) or cot death is a diagnostic entity which can be applied in such a case. It is one of the most common causes of infant deaths world-wide in the postneonatal period, from 1 month to 1 year of age. Features which may assist the clinician to arrive at such a diagnosis are discussed in this paper. Since the first sign of SIDS is death, leaving the clinician with no opportunity to intervene, attention has been focussed on whether or not the infants prone to suffer from this tragic demise can be identified and preventive measures instituted to save them. In this respect, intensive and extensive research efforts over the last two decades have led to what is now commonly referred to as the Apnea Hypothesis for SIDS, and the resultant current intervention method of home apnea monitoring. The apnea hypothesis and other aetiological factors that may help predict infants at risk are discussed. Finally, current management of infants at risk is discussed for completeness since the facilities for such management are not yet in common use in Nigeria and the West African sub-region, and the level of literacy may severely limit the size of the population that could benefit from such management even if the facilities were available. PMID- 12769317 TI - Cardiopulmonary bypass in Jehovah's Witnesses. AB - Jehovah's Witnesses do not accept heterologous blood transfusion for religious reasons. Autologous transfusions are also rejected if there is no continuous contact between the circulation and the autologous blood. There is, therefore, the need to adopt methods which will avoid transfusion of heterologous blood in elective cases as far as Jehovah's Witnesses are concerned. We report two cases where pre-operatively administration of nutritional supplements, haematenics, erythropoietin, antimalarials and the modification of the extra-corporeal circulation bypass circuit allowed successful open-heart surgery using cardiopulmonary bypass. PMID- 12769318 TI - Marfan's syndrome presenting with abdominal aortic aneurysm: a case for vigilance. AB - We present the case of a 16-year old student with Marfan's syndrome and abdominal aortic aneurysm who presented with a diagnostic conundrum. He presented with a three months history of progressive painful left upper abdominal mass and back pain. It became severe in the last two weeks before presentation and was associated with constipation. This mass was thought to be of splenic origin but the initial ultrasound suggested a pancreatic pseudocyst. Review of his previous hospital record revealed that he had been treated for severe myopia which started at infancy. Another opthalmic review at our centre revealed bilateral ectopia lentis. He had no cardiac signs and no family history of cardiovascular diseases. He is the 6th of 8 siblings, all the family members are alive and healthy except one sibling who died at 7 months. The diagnosis of abdominal aortic aneurysm was only made at laparotomy and confirmed by on-table aortogram. He had excision of the aneurysmal sac and replacement with on-lay dacron tube graft. He died on the 4th post-operative day. A diagnosis of abdominal aortic aneurysm was not made at initial presentation because of the rarity of this condition in our environment and incompetence of the ultrasonographer. Aortic aneurysm in Marfan's syndrome is commonly found in the thoracic part of the aorta, however in this case, it is abdominal. A high index of suspicion is necessary to avoid missing this pathology, therefore the need for vigilance. PMID- 12769319 TI - The diagnostic dilemma of intraoperative hyperpyrexia in a malaria endemic area. AB - Malignant hyperthermia (MH) is a serious and feared complication of general anaesthesia. The diagnosis of MH may pose a diagnostic dilemma, as its two most common signs tachycardia and hyperthermia are also common features of infections, transfusion and drug reactions. We report three cases of hyperthermia occurring during anaesthesia posing diagnostic dilemma in which two of the cases appear to be due to malaria. It is therefore suggested that all patients undergoing anaesthesia in a malaria endemic area should be investigated and treated for malaria if indicated preoperatively even when asymptomatic to prevent such diagnostic confusion. PMID- 12769320 TI - Beckwith-Wiedemann Syndrome (BWS): a case report and literature review. AB - Beckwith-Wiedemann Syndrome (BWS), also known as the EMG (Exomphalos, Macroglossia, Gigantism) syndrome was recognised independently by Beckwith in 1963 and Wiedemann in 1964 and is now a well established entity having been reported in more than two hundred individuals. It constitutes a wide spectrum of clinicopathologic entity with varied combinations of congenital and time dependent abnormalities that often make diagnosis and management tasking. There is paucity of report in the literature on this entity from the developing world. We present a case recently seen at the University College Hospital (UCH) Ibadan, Nigeria in order to create further awareness and highlight peculiarity of management as may be applicable in a setting as ours. PMID- 12769321 TI - Human immunodeficiency virus and invasive external otitis--a case report. AB - Acute invasive external otitis is an uncommon life-threatening infection of the external auditory canal (EAC), most often affecting the elderly diabetic patients. Although few reports have been made in HIV-positive/Aids patients among the caucasians. We present here a 25 year old nursing mother with a month history of fever, persistent otalgia with acutely inflammed EAC, gross facial cellulitis, mastoid abscess and facial paresis, following a minor left ear trauma with a matchstick. This unusual course of ear infection in an otherwise healthy young adult prompts a search for an immunodepressing factor which was confirmed to be Human Immunodeficiency Virus (HIV). This article highlights the clinical peculiarities and the management of invasive external otitis in an HIV-positive patient. PMID- 12769322 TI - . . . and who shall benefit? PMID- 12769323 TI - Competency is no reason for complacency. PMID- 12769324 TI - Use of nontraditional therapies--implications for older adults. AB - Using nontraditional therapies to replace traditional medical treatment is becoming more popular in the United States. Nurses in the twenty-first century are faced with the challenge of broadening their view of health care and discovering the benefits of nontraditional, often ancient, health care principles and practices. Health care professionals and patients need to understand, however, that although a nontraditional therapy may be natural or noninvasive, it is not necessarily safe and without consequences. This article discusses the increased use of nontraditional therapies among older adults with a focus on the use and perioperative implications of botanical, herbal, and nutritional supplements. PMID- 12769325 TI - Laparoscopic adjustable gastric banding for morbid obesity. AB - Morbid obesity, with its accompanying human and monetary costs, is a serious and growing health problem worldwide. Surgical treatments are recognized as being effective, but until recently, the available surgical options entailed invasive techniques that permanently altered the gastrointestinal tract. In June 2001, the US Food and Drug Administration approved an adjustable gastric banding system that can be implanted laparoscopically for the treatment of morbid obesity. This article discusses laparoscopic adjustable gastric banding, including patient selection and evaluation, perioperative care, and the importance of a team approach to patient management and follow-up care. PMID- 12769326 TI - What is new with renal transplantation? AB - Dialysis and transplantation bring new hope and life to thousands of patients with end-stage renal disease. Renal transplantation restores reasonably normal health to patients whose kidneys no longer function and frees them from the limitations imposed by dialysis. Improved graft survival rates have further enhanced the desirability of transplantation. Currently in the United States, more than 80,000 people are living with a functioning renal transplant. The introduction of laparoscopic and laparoscopy-assisted techniques has proven to be a major improvement to living donation. This less invasive method of donating a kidney has more than doubled the chance that a patient with kidney failure will receive a transplant from a friend or loved one. New immunosuppressant medications, improved success rates, and the proliferation of transplantation centers have made renal transplantation a viable choice for many patients. The future will be dependent upon a marked increase in organ donation, which in turn will bring about earlier transplantation for patients with end-stage organ failure. PMID- 12769327 TI - Legal and ethical issues related to nonheart beating organ donation. AB - Major breakthroughs in immunosuppressant medication and advances in surgical techniques have increased the number of successful transplantations, but the demand for organs far outweighs the supply. People continue to die while waiting for donor organs to become available. Nonheart beating organ donation is a means of increasing the supply of organs to be transplanted. It is based on cardiac cessation criteria. This article explores the ethical and legal considerations, as well as implications for perioperative nurses. PMID- 12769328 TI - Bipolar transurethral resection of the prostate--a new approach. AB - Benign prostatic hyperplasia (BPH) is a common problem among older men. Transurethral resection of the prostate (TURP) is a common surgical procedure for treatment of BPH. Typically, TURP is performed using monopolar electric current and a wire electrode; however, the irrigating solution used during this procedure has the potential to cause transurethral resection (TUR) syndrome. A new technology uses bipolar electrosurgery, which creates less risk of electrical shock, and saline irrigation, which eliminates the risk for TUR syndrome. PMID- 12769330 TI - Ingredients for a successful pediatric preoperative care process. PMID- 12769329 TI - The cost and efficacy of two wound treatments. AB - Using the physiology of moist wound healing as the framework, this nonexperimental, retrospective chart review compared the rate of wound healing and cost of wound care associated with wet-to-dry normal saline gauze dressings to the rate of wound healing and cost of wound care associated with amorphous hydrogel dressings for patients with infrainguinal arterial disease and diabetes. These patients were discharged from the hospital to home care for management of perioperative arterial surgical wound dehiscence and nonhealing ulcerations. The sample included 25 patients who used wet-to-dry normal saline gauze dressings and 25 patients who used amorphous hydrogel dressings. Repeated measures of analysis of covariance revealed a similar rate of wound healing in the two groups. The overall cost of wound care was significantly higher (P = .006) for patients in the normal saline group, with a higher number and cost of home nursing visits. The cost of supplies was not significantly different between groups, although amorphous hydrogel dressings cost an average of dollar 50 more than wet-to-dry normal saline gauze dressings. The two treatments are equally efficacious in promoting wound healing, but amorphous hydrogel dressings are significantly more cost effective and, thus, a better value for the home care dollar. PMID- 12769331 TI - Information at your fingertips. PMID- 12769332 TI - Mitochondria as a target for inducing death of malignant hematopoietic cells. AB - Mitochondria plays a central role in apoptotic cell death. The intermembrane space of mitochondria contains a number of soluble molecules whose release from the organelle to the cytosol or the nucleus induces cell death. Thus, molecules that directly trigger mitochondria membrane permeabilisation are efficient cytotoxic drugs. Mitochondria is one of the cellular targets for commonly used epipodophyllotoxins, adenine deoxynucleoside analogs and taxanes as well as recently developped agents such as the pentacyclic triterpene betulinic acid and the lymphotoxic agent FTY720. Most informations on anthracyclines point to the mitochondrial membrane as the main target of cardiotoxicity. Mitochondria is also a target for arsenite trioxide, an old cytotoxic agent recently used for treating acute promyelocytic leukemia, lonidamine, a dichlorinated derivative of indazole 3-carboxylic acid developped as a chemosensitizer, the retinoic acid receptor gamma activator CD437 and nitric oxide (NO). Recently, cytotoxic drugs have been specifically designed to directly affect the mitochondrial function. These include the positively charged alpha-helical peptides, which are attracted to and disrupt the negatively charged mitochondrial membrane, thus inducing mammalian cell apoptosis when targeted intracellularly. Various strategies have been proposed also to directly inhibit Bcl-2 and related anti-apoptotic proteins, including antisense oligonucleotides (e.g. Genasense, currently tested in phase III trials), small molecules that mimic the BH3 dimerization domain of these proteins and kinase inhibitors. Ligands of the mitochondrial benzodiazepine receptor such as the isoquinolone carboxamide derivative PK11195 also overcome the membrane-stabilizing effect of Bcl-2, whereas the adenosine nucleotide translocator (ANT) and the mitochondrial DNA are two other potential cellular targets for cytotoxic agents. Potentially, new compounds directly targeting the mitochondria may be useful in treating hematological malignancies. The challenge is now to selectively target these mitochondria permeabilizing agents to malignant cells. This review briefly summarizes the role of the mitochondria in cell death and describes these various strategies for targeting the mitochondria to induce apoptosis. PMID- 12769333 TI - The regulation of hematopoietic stem cell and progenitor mobilization by chemokine SDF-1. AB - The chemokine, stromal derived factor-1 (SDF1) has been shown to modulate the homing of stem cells to its site by mediating chemotaxis. We demontrated overexpression of SDF-1 induced mobilization of cells with stem cell potential (CFU-S, long-term reconstituting cells). Moreover, SDF-1 activated matrix metalloproteinase-9 induced in bone marrow (BM) cells, causing the release soluble Kit-ligand and permitting the transfer of hematopoietic stem cells (HSCs) from a quiescent to a proliferative niche. The role of SDF-1 in the process of HSC migration and the mechanism underlying this HSCs migration will be reviewed. These studies lay the foundation for using SDF-1 to induce mobilization of progenitor cells in a BM transplantation setting. PMID- 12769334 TI - Cellular immunotherapy after autologous hematopoietic stem cell transplantation: experimental strategies and clinical experiences. AB - Recurrence of disease after autologous hematopoietic stem cell transplantation is at least partly due to contamination of the reinfused transplant with tumor cells, thereby limiting the clinical outcome after transplantation. On the other hand, immunological effector cells are capable of purging bone marrow transplants in vitro and of destroying disseminated tumor cells in vivo. Cellular immunotherapy subsequent to autologous stem cell transplantation is therefore expected to have a major impact on recurrence rates of the disease. In this review, we present various strategies utilizing immunologic effector cells for elimination of disseminated tumor cells and discuss the advantages and limitations of cellular immunotherapy after autologous hematopoietic stem cell transplantation. PMID- 12769335 TI - Immunosuppressive treatments for myelodysplastic syndromes. AB - The myelodysplastic syndromes (MDS) comprise a heterogeneous group of clonal hematopoietic stem cell disorders, while, immunological abnormalities are frequently observed in patients with MDS. Several reports revealed that about 10% of MDS patients have clinical autoimmune disorders like skin vasculitis, rheumatic disease, or autoimmune hemolytic anemia. Furthermore, serological immunological abnormalities like hyper- or hypogammaglobulinemia, positivities of antinuclear antibody, positivities of direct Coombs test, or inverted CD4/8 ratios were found in 18-65% of patients with MDS. Recently immunosuppressive therapies including prednisolone, antithymocyte globulin, and cyclosporin A (CsA) are used to treat cytopenia in some patients with MDS. We examined the efficacy of CsA in 50 patients with MDS. Hematologic improvement was observed in 30 (60%) patients especially for erythroid lineage. There were significantly more responders with good karyotype or DRB1*1501 than with intermediate/poor karyotypes or with other HLA types. MDS with erythroid hypoplasia is a rare form of MDS, and has not yet been clearly defined. We reported four patients with MDS with erythroid hypoplasia who had morphological evidence of myelodysplasia and low percentage of erythroid precursors. Rearrangements of the TCR-beta and -gamma genes were seen in these patients using Southern blot and PCR analysis. Also they had skewed TCR usages using TCR repertoire analysis. Their anemia drastically improved with CsA therapy. We have to establish the clinical usefulness of immunosuppressive therapy in MDS patients and simple tools for revealing T-cell mediated myelosuppression in the individual patients for decision-making. PMID- 12769336 TI - New candidate virus in association with Hodgkin's disease. AB - Epidemiologic and molecular investigations of Hodgkin's disease (HD) suggest a strong infectious association. The Epstein-Barr virus (EBV), together with its viral proteins, is expressed in Hodgkin-Reed-Sternberg (HRS) cells in the lymph nodes involved by HD. EBV is more likely to be related to childhood and older adult cases of HD and is much less frequently expressed in young adult HD patients, the group most expected to be associated with an infectious agent. In addition, the "hit and run" theory of EBV infection remains speculative and no other lymphotropic viruses studied to date seem to satisfy the quest for a new candidate virus in young adults with HD. We have recently found preliminary evidence suggesting a possible association between the measles virus (MV) and HD. This evidence is the subject of the present review. PMID- 12769337 TI - The current status of therapy for adult T-cell leukaemia-lymphoma in Japan. AB - More than 25 years have passed since adult T-cell leukaemia-lymphoma (ATLL) was identified as a distinct clinical entity clustered in the southwestern part of Japan. During these years, its causative agent, Human T-cell leukaemia virus type I (HTLV-I), was found, and remarkable clinical variance of this disease was recognized afterward. Because of the regional occurrence of this disease in the world, young medical scientists whose community is not endemic for this disease may have little understanding of it and may think that it a T-lymphoblastic leukaemia-lymphoma of adult-onset. In Nagasaki prefecture in Japan, where HTLV-I carriers account for 3-5% of the population, the incidence of ATLL is almost the same as the incidence of B-cell lymphomas, and hence ATLL is a matter of major concern. In contrast to the vast accumulation of knowledge about the oncogenic role of HTLV-I and molecular biology of ATLL cells, improvement in the prognosis of patients has not been satisfactorily achieved except for some recent progression. Here, we review the current status of therapy for ATLL in Japan and discuss how best to manage this difficult disease at this point and what next step should be taken. PMID- 12769338 TI - Evaluation of imipenem 1.5 g daily in febrile patients with short duration neutropenia after chemotherapy for non-leukemic hematologic malignancies and solid tumors: personal experience and review of the literature. AB - Numerous studies have demonstrated efficacy of imipenem-cilastatin, 50 mg/kg/day, as first line therapy in febrile patients with neutropenia of short duration consecutive to cytostatic chemotherapy. However, only two studies used low dosage of this antibiotic as 1.5 g/day, in prospective, double blind, randomized clinical trials, in this indication. Efficacy and tolerability of imipenem cilastatin 0.5 g three times daily IV in 30-min infusions, as first-line empiric therapy, were retrospectively evaluated in our hematological unit. From January 1996 to September 2000, 30 neutropenic patients (12 females) with 45 febrile episodes were included. Median age was 57.5 years (31-75). Twenty-four of them had lymphomas, 4 solid tumors and 2 myelomas. There were 13 clinically documented infections, (CD, 28.8%), 16 microbiologically documented infections, (MD, 35.6%) and 16 febrile episodes corresponding to fever of unknown origin, (FUO, 35.6%). The median neutrophils count on nadir (n = 44), was 67/mm3 (8-369). The median duration of neutropenia was 5 days (3-15). Bacteremia was observed in 10 patients, urinary tract infection in 3 patients. The most frequently isolated microorganism was Escherichia coli. The overall success rate of the first line therapy was 66.7%. Adverse events were observed in 11.1% of the patients without necessity to stop treatment. The MD infections showed a lower rate of success compared with CD infections and FUO. These data were in accordance with the previous studies. The importance of number of microorganisms (p = 0.007) and of infected sites (p = 0.01) appeared as prognostic factors (univariate analysis). Although imipenem-cilastatin has been used in numerous studies as empiric broad spectrum antibiotic therapy in the treatment of febrile neutropenic cancer patients, the exact dosage of this antibiotic is still not standardized. However, utilization of this antibiotic in monotherapy at low dosage seems to us to be safe and effective as usual dosage in the antimicrobial treatment ofthe febrile patients with post chemotherapy neutropenia of short duration. PMID- 12769339 TI - Current status and future of relapsed primary central nervous system lymphoma (PCNSL). AB - The treatment of primary central nervous system lymphoma (PCNSL) has centered around high-dose methotrexate and radiotherapy (RT). Methotrexate administered intra-arterially (IA) with blood-brain barrier disruption (BBBD) and without RT, has been a highly effective treatment with a 5 year survival of 42% without cognitive loss. The purpose of this analysis is to determine responses for patients with relapsed PCNSL treated with second line IA carboplatin-based chemotherapy with BBBD. Between February 1991 and April 2000, 37 relapsed PCNSL patients, most who failed front line therapy with methotrexate based chemotherapy, were treated at Oregon Health & Science University (OHSU) and Hadassah Hebrew University Hospital (HHUH) with IA carboplatin-based chemotherapy with BBBD. Nine patients had prior RT. The mean age was 57.5 years, and all but 1 patient were treated within 8 months after relapse. The median time for survival from first IA carboplatin/BBBD treatment was 6.8 months;however, 7 out of 37 patients survived > or = 27 months. Nine patients had radiographic complete response (CR), 4 patients had radiographic partial response (PR), 12 had stable disease (SD), 10 had progressive disease (PD), and 2 were non-evaluable. The median time to failure for patients with CR and PR was 9.1 months. One long-term survivor is alive at 91.0 months from first carboplatin/BBBD treatment. In conclusion, we show that relapsed PCNSL has shown sensitivity to second line IA carboplatin-based chemotherapy with BBBD. We have developed a new protocol using i.v. rituximab prior to BBBD with IA carboplatin, i.v. cyclophosphamide and i.v. etoposide phosphate. The long-term program goal is to consolidate dose-intensive chemotherapy with monoclonal antibody directed radiation. Because patients with recurrent PCNSL commonly continue to relapse even after obtaining a complete response to enhanced chemotherapy treatment, patients w ho complete or fail the above carboplatin/BBBD treatment regimen will be offered consolidation with radioimmunotherapy using zevalin (Ibritumomab tiuxetan), IDEC-2B8 conjugated with yttrium-90 (90Y). PMID- 12769340 TI - A randomized trial comparing intensified CNOP vs. CHOP in patients with aggressive non-Hodgkin's lymphoma. AB - The standard CHOP regimen may cure 30-40% of patients with advanced aggressive non-Hodgkin's lymphoma (ANHL). Mitoxantrone is an anthracenedione, which is active in NHL and its toxicity profile may be more favorable than doxorubicin with respect to alopecia, mucositis and cardiotoxicity. This study was designed to compare the effectiveness of an escalated dose of mitoxantrone with that of standard doxorubicin, used in the CHOP regimen in patients with ANHL. One hundred and forty three eligible patients with ANHL were randomized to receive 6 cycles of either CHOP (n = 71) or intensified CNOP (iCNOP) (n = 72), with mitoxantrone 20 mg/m2, i.v., d.1 instead of doxorubicin. Complete responders (CR) were again randomized either to receive interferon-alpha (IFN-alpha) maintenance (3 MU t.i.w., s.c.) or not. The CR rate was 70 vs. 76% for iCNOP and CHOP (p = 0.45), and the overall response rate was 81 vs. 83%, respectively (p = 0.71). The 5-year failure free survival (FFS) was 48 and 50% in the iCNOP and CHOP arm, respectively (p = 0.45), and the 5-year overall survival (OS) was 61 vs. 64% (p = 0.56). IFN-alpha did not prolong relapse free survival (p = 0.91). iCNOP produced less alopecia (p = 0.001) but more febrile episodes (p = 0.04) than CHOP, while requiring more frequent G-CSF support (p = 0.01). Two cases of acute myelogenous leukemia (AML) were recorded, both in the iCNOP arm (p = 0.14). In conclusion, iCNOP was equally effective to CHOP in patients with ANHL, producing more leukopenia and febrile episodes, but less alopecia. The development of two cases of secondary AML in th e iCNOP arm is of concern. PMID- 12769341 TI - Immunoglobulin gene rearrangement investigations in the diagnosis of lymphoid malignancies from formaldehyde-fixed biopsies. AB - Determination of the biologic potential of lymphoid proliferations in biopsies can be difficult by standard histological or even immunohistochemical examination. Polymerase chain reaction (PCR) has been used with increasing frequency to detect clonal rearrangements of the immunoglobulin heavy chain (IgH) in formaldehyde fixed, paraffin wax embedded tissues. Sensitivity ranges between 50 and 80%, and therefore at least 20% of neoplasms remain undetected by these approaches. Few investigators have attempted to detect immunoglobulin light chain (IgL) gene rearrangements by PCR using paraffin wax embedded samples. We studied 29 cases of B-cell neoplasms, along with 21 cases with equivocal histology and 4 reactive biopsies, using degenerate oligoprimers to amplify Ig(kappa) and Ig(lambda) light chain genes, along with IgH (Fr 1, 2 and 3) gene rearrangement analysis. The combination of these methods detected clonality in 93% of cases (27/29) with histological diagnosis of B-NHL. Fr2 and Fr3 primers detected clonality in 79% (23/29) of cases. IgL chain rearrangements detected 4 cases (14%), negative for IgH rearrangements, improving sensitivity from 79 to 93%. Clonality was detected in 52% (11/21) of histologically equivocal lymphoid proliferations, including one case detected by IgL rearrangements which was negative for IgH rearrangements. Archival material from 4 cases with reactive histology produced polyclonal results. These results confirm that PCR based immunoglobulin gene rearrangement is a sensitive and specific method for demonstrating B-cell clonality in paraffin-wax embedded sections. The addition of IgL analysis to the IgH assay allows the detection of greater than 90% of B-cell lymphoproliferative disorders from routine histological specimens with poor preservation of genomic DNA. PMID- 12769342 TI - Vaccination against infections in chronic lymphocytic leukemia. AB - Chronic lymphocytic leukemia (CLL) is a well-defined mature B-cell neoplasm associated with increased susceptibility to infections. Two major options in prevention of infections in CLL, intravenous gammaglobulin treatment and antimicrobial chemoprophylaxis, have not resulted in satisfactory outcome. A third strategy, antimicrobial vaccination, is the topic of this minireview. We collected articles and their references concerning CLL vaccination from the Medline database starting from 1966 and thirteen relevant studies were found. Plain bacterial polysaccharide vaccines would seem to be ineffective in antibody formation in patients with CLL. However, protein and conjugate vaccines appear to be more immunogenic and their responses may be further enhanced with ranitidine adjuvant treatment. New well-designed investigations are needed to develop appropriate vaccination strategies and evaluate vaccination efficacy in infection morbidity and mortality in CLL. PMID- 12769343 TI - Clinico-hematologic features of myelodysplastic syndrome presenting as isolated thrombocytopenia: an entity with a relatively favorable prognosis. AB - The existence of isolated cytopenia in myelodysplastic syndrome (MDS) has been described, however, the exact clinico-hematologic features of such MDS patients are still obscure. The aim of this study was to provide additive clinico hematologic information on MDS patients with isolated thrombocytopenia in comparison with idiopathic thrombocytopenic purpura (ITP). We searched for MDS with isolated thrombocytopenia in 146 sequential patients with MDS and evaluated their clinical features at the time of MDS diagnosis. We found 13/146 (8.9%) patients with MDS showing isolated thrombocytopenia. These patients were male predominant (10:3) and were all diagnosed as refractory anemia after reassessment of marrow findings, however, two of them had an initial diagnosis of ITP. Leukemic transformation was rarely noted (1/13 patients), but 1 patient developed myelofibrosis. Cytogenetic study demonstrated that 3 patients had del(20q), 2 had t(1;7)(q10;p10), and 5 showed normal karyotypes. The most prominent morphologic feature in the megakaryocytes was the presence of micromegakaryocytes (5/13) and 8/13 had hypogranulated neutrophils, whereas pseudo-Pelger nuclear anomaly was rarely detectable. Of note is that 7/13 patients had an increased number of megakaryocytes in the marrow. Most patients survived for more than 2 years. Approximately 9% of MDS patients showed isolated thrombocytopenia and most of them had a favorable prognosis. Some MDS patients with isolated thrombocytopenia have been mistakenly diagnosed as having ITP, since approximately 50% of our MDS patients with isolated thrombocytopenia had an increased number of megakaryocytes with low grade dysplasia. Therefore, careful attention to differential diagnosis is recommended for these patients. PMID- 12769344 TI - Lineage-specific chimaerism quantification after T-cell depleted peripheral blood stem cell transplantation. AB - Patients that receive a T-cell depleted (TCD) hematopoietic stem cell transplantation (SCT) show higher risk of graft failure/rejection and of disease relapse than those that receive unmanipulated grafts. The purpose of the present investigation was to analyze the usefulness of chimaerism quantification in bone marrow (BM), peripheral blood (PB), and leukocyte lineages such as T lymphocytes (CD3+,both CD4+ and CD8+), B lymphocytes (CD19+) and myeloid cells (CD15+), for the early detection of graft failure/rejection episodes and disease relapse after TCD-PBSCT. Two of the ten (2/10) patients included in the study showed stable complete chimaerism (CC). The other 8/10 patients showed decreasing mixed chimaerism (MC) and 7 of them had either graft failure (n = 1)/rejection (n = 3) or disease relapse (n = 3). In two patients relapsed from chronic myeloid leukemia, MC was observed in BM and PB, with higher percentages of autologous cells in BM, as well as in leukocyte lineages, with higher percentages of recipient cells in the myeloid lineage than in lymphocytes. Combined analysis of chimaerism and minimal residual disease allowed early diagnosis of relapse and successful rescue therapy with donor leukocyte infusions (DLI), before the onset of hematological relapse. Chimaerism analysis allowed early diagnosis of incipient graft rejection in 3 patients. These patients showed MC both in BM and PB, with greater percentages of recipient cells in PB. Analysis of leukocyte lineages showed higher percentages of autologous cells in T lymphocytes (mainly CD8+) than in B or myeloid cells. Two of these patients were successfully treated with DLI and recovered normal PB counts and BM cellularity, as well as CC. The graft versus recipient hemopoiesis effect harbored by the donor immunocompetent cells infused seems useful forthe treatment of graft rejection, provided that an early diagnosis is made. PMID- 12769345 TI - Adjusted conditioning for allogeneic transplantation in a single center setting: mixed chimerism heralds relapse. AB - The role of mixed chimerism on subsequent relapse was prospectively evaluated in an allotransplant program. Sixty-six patients with median age of 54 and mainly high-risk hematologic disease and/or solid tumors had individually adjusted non myeloablative conditioning. Thirty-nine donors were siblings and 27 unrelated. Frequent chimerism analyses supported immune manipulation including donor lymphocyte infusions. The need for transfusions, i.v. fluids, and antibiotics, and weight loss was less than in a control cohort. Most patients had immediate full and consistent donor chimerism; one-third required immune manipulation. Eight of ten evaluable CML patients were BCR/ABL-negative at days 14-58 post transplant. Mixed chimerism frequently preceded relapse, and the relapse rate was 38% in 26 patients with mixed chimerism vs. 11% among 35 with consistent full donor chimerism (p = 0.015). The current transplant- and disease-related mortality were 11 and 9%, respectively, among 35 non-high-risk patients, and 35 and 10% for 29 high-risk patients with hematologic malignancy. With a median follow-up of 15 months the 2-year overall survival is 73% for non-high-risk, and 46% for high-risk patients. Adjusted conditioning reduces early toxicity and resource requirements without impairing tumor control, probably due to a rapid establishment of the graft-versus-cancer effect. Mixed chimerism heralded relapse, and tumor-related mortality is not greater with adjusted than with conventional conditioning. PMID- 12769346 TI - Specific destruction of hybridoma cells by antigen-toxin conjugates demonstrate an efficient strategy for targeted drug therapy in leukemias of the B cell lineage. AB - Many types of leukemia including multiple myeloma remain essentially incurable despite recent developments in immuno- and chemotherapy. The effectiveness of these therapies might be greatly enhanced by targeting cell surface proteins unique to the malignant clone, which for leukemias of the B cell lineage means clonotypic surface immunoglobulin (sIg). As this immunoglobulin (Ig) is necessarily epitope specific, we are developing ligand-toxin conjugates (LTCs) as a strategy for delivering toxins and other drugs to clonotypic tumor cells. Here we report in vitro studies that illustrate the effectiveness of this approach. LTC comprising the DNP hapten conjugated to ricin A toxin (DNP-RTA) were shown to specifically and effectively kill anti-DNP secreting murine hybridoma (U7.6) cells but not other hybridoma cells (1B12), a murine erythroleukemia cell line (Friend's Leukemia or) normal mouse spleen cells. In addition to direct toxicity, LTC treatment negatively affected the growth characteristics of the few surviving cells as reflected in decreased growth index and an increase in growth inhibition over 72 h post treatment. Interestingly, U7.6 cells that survived one or two LD90 dose(s) of LTC showed no alteration in their dose response to a subsequent attack of LTC indicating that this treatment strategy may not induce drug resistance. These data suggest that LTC therapy may be a new and effective strategy for specific destruction of tumor cells such as myeloma plasma cells and could be extended to other tumors where clonotypic receptors can be identified. PMID- 12769347 TI - Retinoic acid induction of CD38 antigen expression on normal and leukemic human myeloid cells: relationship with cell differentiation. AB - Differentiation in the hematopoietic system involves, among other changes, altered expression of antigens, including the CD34 and CD38 surface antigens. In normal hematopoiesis, the most immature stem cells have the CD34 + CD34 - phenotype. In acute myeloid leukemia (AML), although blasts from most patients are CD38 +, some are CD38 - . AML blasts are blocked at early stages of differentiation; in some leukemic cells this block can be overcome by a variety of agents, including retinoids, that induce maturation into macrophages and granulocytes both in vitro and in vivo. Retinoids can also induce CD38 expression. In the present study, we investigated the relationship between induction of CD38 expression and induction of myeloid differentiation by retinoic acid (RA) in normal and leukemic human hematopoietic cells. In the promyelocytic (PML) CD34 - cell lines, HL60 and CB-1, as well as in normal CD34 + CD34 - hematopietic progenitor cells RA induced both CD38 expression as well as morphological and functional myeloid differentiation that resulted in loss of self-renewal. In contrast, in the myeloblastic CD34 + leukemic cell lines, ML-1 and KG-1a, as well as in primary cultures of cells derived from CD34 + -AML (M0 and M1) patients, RA caused an increase in CD38 + that was not associated with significant differentiation. Yet, long exposure of ML-1, but not KG-1, cells to RA resulted in loss of self-renewal. The results suggest that while in normal hematopoietic cells and in PML CD34 - cells induction of CD38 antigen expression by RA results in terminal differentiation along the myeloid lineage, in early myeloblastic leukemic CD34 + cells, induction of CD38 and differentiation are not functionally related. Since, several lines of evidence suggest that the CD38 - cells are the targets of leukemic transformation, transition of these cellsinto CD38 + phenotype by RA or other drugs may have therapeutic effect, either alone or in conjunction with cytotoxic drugs, regardless the ability of the cells to undergo differentiation. PMID- 12769348 TI - Mutations in the retinoblastoma-related gene RB2/p130 in adult T-cell leukaemia/lymphoma. AB - The retinoblastoma (Rb) family consists of the tumor suppressor Rb/p105 and related proteins p107 and Rb2/p130. Although the involvement of the RB/p105 gene in Adult T-cell leukaemia/lymphoma (ATL) has been studied, no mutational data is reported regarding the RB2/p130 gene in ATL. We screened for mutations of the RB2/p130 gene. Mutation was detected in 1 of 41 primary ATL sample. This is the first report describing mutation of the RB2/p130 gene in ATL, suggesting that RB2/p130 may be involved in the development of ATL, and may behave as a tumor suppressor gene in T lymphocytes. PMID- 12769349 TI - Non-myeloablative hematopoietic stem cell transplantation for the treatment of adult T-cell lymphoma in a patient with advanced hepatic impairment. AB - A 59-year-old man with liver cirrhosis due to hepatitis B virus infection received non-myeloablative stem-cell transplantation (NST) for the treatment of adult T-cell lymphoma. The preparative regimen consisted of cyclophosphamide and fludarabine. While the pharmacokinetics of these drugs was altered in this patient, his clinical course was uneventful without the development of severe hepatic damage. Complete remission was achieved on day 56. Although he finally died of hemorrhage from esophageal varices on day 68, this case suggests that ATL may be a good candidate for allogeneic HSCT, and that NST may be feasible for patients with hepatic impairment. PMID- 12769350 TI - Acute myelogenous leukemia complicated by acute necrotizing ulcerative gingivitis due to Aspergillus terreus. AB - Infections caused by Aspergillus terreus are rare but have been associated with a poor outcome in immunocompromised patients due to frequent resistance to conventional antifungal therapy. This report describes a case of a woman who developed acute necrotizing ulcerative gingivitis (ANUG) due to A. terreus during induction chemotherapy for acute myelogenous leukemia. She initially failed to respond to treatment with amphotericin B but the infection resolved following the introduction of oral itraconazole. Opportunistic infections caused by A. terreus are an emerging problem and can be associated with a high mortality rate. Early microbiological diagnosis is critical since resistance to amphotericin B is likely and itraconazole appears to be an effective treatment for this infection. PMID- 12769351 TI - T cell lymphoma associated with myelofibrosis. AB - Myelofibrosis is most frequently associated with certain primary myeloproliferative disorders,but is rare in lymphoid neoplasms. We report the fourth case associated with T-cell lymphoma, involving bone marrow, lymph nodes and spleen. Marked extramedullary hematopoiesis was present. Myelofibrosis subsided completely with response to standard anti-lymphoma combination chemotherapy. Since lymphomatous splenic and bone marrow involvement was minimal in our patient, fibrotic bone marrows should be carefully evaluated for lymphoma. PMID- 12769352 TI - Asymptomatic pericardial effusion in chronic myelogenous leukemia. AB - In chronic myelogenous leukemia (CML), pericardial effusions are rarely seen. Pericardial tamponade may occur in many of the patients with pericardial effusion. Here in a 45-year-old male with newly diagnosed chronic phase of Philadelphia chromosome (the Ph)-positive CML, asymptomatic pericardial effusion was detected by echocardiography. The pericardial effusion disappeared after chemotherapy without pericardiocentesis, within six months. Pericardial effusion should always be suspected in patients with chronic phase of CML, even though they are asymptomatic. In our case the pericardial effusion was probably due to extramedullary haematopoiesis because it improved with chemotherapy. Follow-up with echocardiography was sufficient in this case. PMID- 12769354 TI - Remission of follicular non-Hodgkin's lymphoma with denileukin diftitox (ONTAK) after progression during rituximab, CHOP and fludarabine therapy. AB - Denileukin diftitox (ONTAK) is indicated for the treatment of patients with cutaneous T cell lymphoma. Clinical experience with this drug in other lymphomas is limited. This case report concern a patient with stage IV follicular lymphoma who, relapsing after autologous transplant and having failed multiply systemic therapies, including retuximab and CHOP, achieved a prolonged remission with denileukin diftitox. PMID- 12769353 TI - B-cell marker negative (CD7+, CD19-) Epstein-Barr virus-related pyothorax associated lymphoma with rearrangement in the JH gene. AB - Pyothorax-associated lymphoma (PAL) develops decades after receiving artificial pneumothorax for pulmonary tuberculosis. The lymphomas, develop in tissue affected by long-standing severe inflammatory process. Most cases demonstrate diffuse large B-cell lymphoma. We present a patient with T-cell phenotype positive and B-cell phenotype-negative (CD7+, CD43+, CD19-, and CD20-) PAL. Southern blot hybridization using immunglobulin heavy chain J region (IgH) gene probe revealed a monoclonal rearrangement, and hybridization using T-cell receptor beta chain (TCR) gene probe revealed a germline configuration. This indicates that the tumor origin was of B-lymphocytes. Chromosomal abnormality of the lymphoma was complicated. It suggested that many transformations occurred. In the transformation process, probably B-cell antigens were lost, and T-cell antigens were aberrantly expressed. PMID- 12769355 TI - Double Philadelphia chromosomes-positive acute lymphocytic leukemia. PMID- 12769356 TI - Bilateral primary breast lymphoma: a case of local recurrence. PMID- 12769357 TI - Primary adrenal lymphoma. PMID- 12769358 TI - Ion-transfer voltammetry at a polarized room-temperature molten salt/water interface. AB - Tetraoctylammonium cation forms a room-temperature molten salt (RTMS) with 2,4,6 trinitrophenolate anion. The RTMS is immiscible with water (W) and forms a stable RTMS/W interface. It has been shown that the RTMS/W interface can be electrochemically polarized. A well-defined voltammetric wave due to the transfer of thiocyanate ion across the RTMS/W interface was observed within the potential window. This is the first example of a polarized RTMS/W interface. PMID- 12769359 TI - Sensing phenothiazine drugs at a gold electrode co-modified with DNA and gold nanoparticles. AB - DNA and gold nanoparticles are co-immobilized at a gold electrode through elaborate self-assembly processes. This configuration has proven to be useful as a sensor for phenothiazine drugs, taking advantage of the well-known, relatively large surface area of gold nanoparticles and the strong intercalation between dsDNA and phenothiazine drugs. This modified electrode has demonstrated good sensitivity and stability towards the oxidation of two model phenothiazine drugs: promethazine and chlorpromazine. A linear dependence between the concentration of phenothiazine drugs and the peak current is observed, with a concentration range of 2.0 x 10(-5)-1.6 x 10(-4) M and 1.0 x 10(-5)-1.2 x 10(-4) M, and a detection limit of 1.0 x 10(-5) M and 7.0 x 10(-6) M, for promethazine and chlorpromazine, respectively. PMID- 12769360 TI - Spectrophotometric assay of yeast vitality using 2,3,5,6-tetramethyl-1,4 benzoquinone and tetrazolium salts. AB - A method for the spectrophotometric assay of yeast vitality was developed using 2,3,5,6-tetramethyl-1,4-benzoquinone and tetrazolium salts. The metabolic efficiency of 2,3,5,6-tetramethyl-1,4-benzoquinone by yeast cells was used as an index of yeast vitality. 2,3,5,6-Tetramethyl-1,4-benzoquinone was reduced to 2,3,5,6-tetramethyl-1,4-hydroquinone by yeast cells. Then, the superoxide anion radicals generated from O2 by reduction with 2,3,5,6-tetramethyl-1,4-hydroquinone under alkaline conditions reduced tetrazolium salts to formazan, which exhibited absorbance maxima at 440 nm. A linear relationship between the absorbance and viable cell density was obtained in the range of 1.0 x 10(5)-2.0 x 10(7) cells/ml for a sample solution. During the cultivation of yeast cells, the absorbance showed almost an anti-parallel change with that of glucose in yeast growth and fermentation, suggesting that the absorbance change reflected the vitality of yeast cells. PMID- 12769361 TI - Effects of fabrication parameters on the enzyme loading and sensor response of enzyme-carrying conductive polymer electrodes. AB - Hydrogen peroxide sensors where horseradish peroxidase (HRP) is incorporated into pyrrole/3-alkylsulfonate pyrrole copolymer films deposited on an SnO2 electrode (HRP/Py-PS electrode) were investigated with regard to the effects of the fabrication parameters (electropolymerization charge, deposition current density, and electrodeposition solution pH) on the amount of surface-immobilized enzyme and the sensor response. The amount of incorporated enzyme was determined with a method recently developed by ourselves. The results suggest that the amount of entrapped enzyme increases almost linearly with the total charge passed, and strongly depends on the polymer film growth rate and the electropolymerization pH. These findings open up a way to control the amount of enzyme and the resultant response of the biosensor by modifying the preparation conditions. PMID- 12769362 TI - Rapid method for the determination of trace fluoride and activation of ion selective electrode. AB - A method for activating ISE is proposed that can allow determination of the fluoride concentration at ng mL(-1) level with good precision and accuracy. Fluoride ISE is activated in 0.5 mol L(-1) HClO4 medium and then fluoride is determined in the same medium. The linear range for the determination of fluoride is between 1.00 x 10(-2)-1.00 x 10(-7) mol L(-1), and the detection limit of the method is 1.0 ng mL(-1). The advantage of this method is that it is free from the use of TISAB solution while being, time-saving and labor-saving. A mechanism study of the activation of FISE in HClO4 medium is explained. The method has been used for the determination of trace fluoride in milk and flour with satisfactory results. PMID- 12769363 TI - Novel ibuprofen potentiometric membrane sensors based on tetraphenylporphyrinato indium(III). AB - Two novel potentiometric membrane sensors responsive to the ibuprofen drug have been developed. These incorporate poly(vinyl chloride) and polyurethane matrix membranes containing 5,10,15,20-tetraphenylporphrinato (TPP) indium(II) ionophore plasticized with dibutylsebacate. The sensors show a near-Nernstian response with anionic slopes of -53 and -55 mV decade(-1), over the concentration range of 4.2 x 10(-6)-1.0 x 10(-2) and 3.3 x 10(-6)-1.0 x 10(-2) M ibuprofen within pH ranges of 4-9 and 5-9 for PVC and PU matrix membranes, respectively. A sensor based on a polyurethane membrane displays a lower detection limit and a wider linear working range, and a sensor based on a PVC membrane exhibits a better overall selectivity, especially in the presence of lipophilic organic anions. Both sensors are used for the quantification and quality-control assessment of ibuprofen in pharmaceutical preparations. The average recoveries are 99.1+/-0.3% and 99.3+/-0.3% for TPP In(III)-PVC and TPP In(III)-PU based membrane sensors, respectively. High selectivities towards ibuprofen in the presence of many anions, drug excipients and diluents are offered by both sensors, which exhibit a non-Hofmeister selectivity pattern. PMID- 12769364 TI - Voltammetric method based on an ion-pairing reaction for the determination of trace amount of iodide at carbon-paste electrodes. AB - Carbon-paste electrodes (CPEs) were studied to elucidate the cathodic stripping voltammetric determination of iodine. At an accumulation potential of 1.0 V (vs. SCE), iodide was preconcentrated on CPEs via an ion-pairing reaction, followed by oxidation to iodine. Then a linear scan voltammogram was recorded after 10 s of quiescent time by a cathodic potential scan from 0.7 V to 0.1 V at a scan rate of 100 mV s(-1). A cathodic peak current was obtained at about 0.38 V. Various experiment parameters such as the acidity, chloride concentration, accumulation potential, accumulation time, concentration of cetyltrimethylammonium bromide (CTAB) and scan rate, were optimized to analyze the iodide by employing linear scan stripping voltammetry. Under the optimal conditions, calibration curves were obtained over a wide concentration range of the iodide ion from 8 x 10(-9) mol L( 1) to 5 x 10(-6) mol L(-1) with a detection limit of 2 x 10(-9) mol L(-1) at an accumulation time of 3 min. The effect of interfering species was evaluated and the procedure was applied to an iodide analysis in table salt, with good results. PMID- 12769365 TI - A new lead(II)-selective PVC-coated graphite rod electrode based on a Schiff base complex. AB - A new lead(II)-selective electrode has been developed based on bis(acetylacetone) p-phenylenediamine-lead(II) [LPb(NO3)2]H2O complex ionophore as a sensing material, dioctylphthalate (DOP) as a solvent mediator and PVC as a matrix. This electrode exhibits a linear Nernstian response over the concentration range of 1 x 10(-5)-1 x 10(-1) mol l(-1) of Pb(II) cation, with a cationic calibration slope of 30.0 +/- 0.2 mV/concentration decade and a detection limit of 2 x 10(-6) mol l(-1) (0.40 ppm). It has a fast response time and can be used for a period of 2 months without any divergence in potentials. The proposed electrode reveals a good selectivity for Pb(II) over a wide variety of other tested cations and could be used in the pH range of 4-8. It was successfully used for direct determination of Pb(II) concentration in some samples. The obtained results show a good agreement with those obtained by an atomic absorption spectrometric method. The average recovery obtained is 96.5 +/- 0.5% with standard deviation of 1.2% (n = 8). PMID- 12769367 TI - Flow-injection analysis method for the determination of nitrite and nitrate in natural water samples using a chemiluminescence NOx monitor. AB - A flow-injection analysis method for the determination of nitrite and nitrate in natural water samples has been developed that consists of two systems based on their reduction to NO with hydrazine and/or ascorbic acid, followed by chemiluminescence detection. The procedure of sweeping the generated NO into an NOx monitor, by means of a gas-liquid separating coil consisting of microporous polytetrafluoroethylene (PTFE) tubing, offers practical advantages. The adjustment of the carrier gas-flow rates could yield the same calibration graphs for the two measurement systems, and the accumulation sweeping mode provides a higher sensitivity. Chemiluminescence detection allows a wide linear calibration range of 5 x 10(-8) to 5 x 10(-5) M. The detection limits for nitrate and nitrite, defined as three-times the standard deviation of measurement blanks, are 2 x 10(-8) M and 1 x 10(-8) M, respectively, and the average precision was 3.2% at ambient natural concentration levels. Recovery tests were between 94% and 106% for a variety of natural water samples. The method is relatively free from interferences from the substances normally found in natural water, and only ferric ion has an effect for the nitrite determination. PMID- 12769366 TI - Determination of dipyrone in pharmaceutical products by flow injection analysis with potentiometric detection. AB - This work describes an FIA potentiometric procedure for the quantification of dipyrone in pharmaceutical products. For the detector, a tubular electrode comprising a polymeric membrane containing tetraoctylammonium as an electroactive material (5% w/w), dibutylphtalate as a mediator solvent (65% w/w) and PVC (30% w/w) directly applied above a graphite conductor support was used. This unit was incorporated into a monochannel FI-system with a 0.1 mol/L phosphate buffer solution (pH = 5.2) as the carrier solution. The electrode showed a linear response from 8.0 x 10(-4) to 10(-1) mol/L dipyrone, a slope of 62.1 +/- 0.2 mV/dec in pH 5.2 units, an injection volume of 500 microL and a carrier flow-rate of 6 mL/min. This procedure was applied to the analysis of pharmaceutical formulations (oral and injectable) containing dipyrone; the obtained results gave a relative error of less than 3.9% and coefficients of variation less than 1% and 5%, respectively, for the FIA and classical iodometric methods. PMID- 12769368 TI - Analysis of acetamiprid in vegetables using gas chromatography-tandem mass spectrometry. AB - A new analytical method has been validated for determining the insecticide acetamiprid in vegetables using gas chromatography (OC) and different mass spectrometric detection techniques, such as full-scan mass spectrometry (MS), and tandem mass spectrometry (MS/MS). For this purpose, a previous extraction of the vegetable sample was carried out with ethyl acetate. In GC-MS/MS, the lowest detectable concentration was 0.001 mg kg(-1), the average recovery rates at various fortification levels (0.015 and 0.030 mg kg(-1)) ranged between 82.4 and 85.7% and the relative standard deviations were lower than 12.2% in all cases. PMID- 12769369 TI - Specific extraction of chromium as tetrabutylammonium-chromate and spectrophotometric determination by diphenylcarbazide: speciation of chromium in effluent streams. AB - A very specific, selective, simple, and inexpensive procedure was developed for the speciation of CrVI and CrIII. This method is based on the quantitative extraction of chromate and CrIII (previously oxidized to CrVI) as a tetrabutylammonium-chromate ion-pair in methyl isobutyl ketone (MIBK), and then back extraction and preconcentration with an acidic diphenylcarbazide (DPC) solution. Back extraction was applied to achieve further preconcentration by a final factor of 20. The CrVI-DPC complex was determined in back-extract by a spectrophotometer at 548 nm. Under these extraction conditions, most of the probable concomitant cations and anions remained in the first inorganic phase. The calibration curve was linear up to 0.14 microg L(-1) of CrVI with a detection limit of 2.22 ng L(-1). The developed procedure was found to be suitable for the determination of the CrVI and CrIII species in various natural water samples with a relative standard deviation of better than 1.6%. The method was successfully applied to the speciation of chromium in spiked natural water samples, and also samples of effluent from a leather treatment plant. PMID- 12769370 TI - Evaluation of beta-cyclodextrin bonded silica as a selective sorbent for the solid-phase extraction of 4-nitrophenol and 2,4-dinitrophenol. AB - A beta-cyclodextrin bonded silica was synthesized by using a convenient method, and was evaluated as a selective sorbent for the solid-phase extraction of 4 nitrophenol and 2,4-dinitrophenol. When double-distilled water was used as the sample matrix, the sorbent showed a strong capacity to adsorb 4-nitrophenol and 2,4-dinitrophenol; the recoveries were found to be 96% and 99%, respectively, with a 1 L water sample. The selectivity of the sorbent was investigated by using a washing step with methanol. Most of the phenols were washed out with 5 mL of methanol, while 4-nitrophenol and 2,4-dinitrophenol still gave recoveries of 94% and 90%. The solution for efficiently eluting the analytes was optimized and the effect of the inorganic salt on the extraction was examined. In order to investigate the potentiality of the sorbent in dealing with real water samples, water from Donghu lake (Wuhan, China) spiked with nine phenolic compounds at microgram per liter levels were preconcentrated on this cartridge. PMID- 12769371 TI - Preparation of uniformly sized molecularly imprinted polymers for phenolic compounds and their application to the assay of bisphenol A in river water. AB - Uniformly sized molecularly imprinted polymers, which can recognize bisphenol A (BPA), have been prepared by a multi-step swelling and polymerization method using BPA or a structurally related analogue of BPA [p-t-octylphenol (OP) or p-t butylphenol (BP)] as the template molecule, 4-vinylpyridine as the functional monomer, and ethylene glycol dimethacrylate as the cross-linker. The BP-imprinted polymer showed higher molecular recognition ability for BPA than the OP-imprinted polymer. The BPA- and BP-imprinted polymers were applied for the assay of a trace amount of BPA in river water using column-switching HPLC with fluorescence detection: A BPA-imprinted polymer was used for removal of BPA from the pretreatment eluent as the trap column, and a BP-imprinted polymer was used for selective pretreatment and enrichment of BPA in river water as the pretreatment column. The calibration graph, constructed from peak area data plotted versus BPA concentration, was linear with a correlation coefficient of >0.999 in the concentration ranges of 25-1000 ppt. The limit of quantitation was 25 ppt with a 5-ml injection. The column-switching HPLC system was successfully applied for the assay of BPA in river water. PMID- 12769372 TI - Ion-exchange properties of colloidal particle consisting of polyaniline and poly(vinyl alcohol) fixed on silica-gel powder. AB - A colloidal powder was prepared by fixing polyaniline (PANI, conducting polymer), poly(vinyl alcohol) (PVA, surfactant stabilizer) and a suitable dopant anion to silica-gel powder. This hydrophilic composite colloidal particle incorporates anions with the protonation of PANI in an acidic solution. The anion can be exchanged with other anions when the colloid is immersed in an acidic solution. Thus, the PANI colloid works as an ion exchanger. The ion-exchange properties on the composite colloidal powder were investigated. Anions were successfully and easily exchanged in the order Br- < Cl- < NO3- < ClO4- < SCN-. This ion-exchange selectivity corresponds largely to the ion-exchange equilibrium constants, which are based on a hydrophobic interaction between the anion and colloid. However, this ion-exchange selectivity does not agree simply with the lipophilic order, but is instead explainable by a gap in the effective ion-exchange capacity due to a size effect between the micropore on the colloidal particle formed by the dopant anion in polymerization and anion sizes in the hydrophobic environment. PMID- 12769373 TI - Serially coupled capillary columns supercritical fluid chromatography with a coupling restrictor. AB - A serially coupled column system for capillary supercritical fluid chromatography was constructed by connecting two capillary columns of different polarities using or without using a coupling restrictor. The influence of the column sequence, the size of the coupling restrictor and the column temperature on the polarity of the system were studied. The system without a coupling restrictor (the directly coupled system) provided an intermediate polarity between two columns, depending on the column geometry, such as the film thickness and column length, although it slightly shifted to that of the first column. In the system with a coupling restrictor, the contribution of the second column to the overall retention increased with its resistance. The use of a coupling restrictor allowed to control the polarity of the coupled system virtually over the whole range between two columns. The temperature was an additional effective parameter for tuning the polarity of the system. PMID- 12769374 TI - Catalytic flow-injection determination of sub-ppb copper(II) using the redox reaction of cysteine with iron(III) in the presence of 2,4,6-tris(2-pyridyl) 1,3,5-triazine. AB - A kinetic-catalytic spectrophotometric flow-injection method was developed for the rapid and sensitive determination of trace amounts of copper(II). The method is based on the catalytic effect of copper(II) on the redox reaction of cysteine with iron(III). Iron(II) produced by the catalytic reaction reacts with 2,4,6 tris(2-pyridyl)-1,3,5-triazine (TPTZ) to form the iron(II)-TPTZ complex (lambda(max) = 593 nm). By measuring an absorbance of the complex, one could determine 0.05-8 ppb copper(II) with the relative standard deviations (n = 10) of 1.6%, 1.3%, and 0.8% for 0.5 ppb, 1 ppb, and 2 ppb copper(II), respectively. The limit of detection (S/N = 3) was 0.005 ppb. The sample throughput was 30 h(-1). The proposed method was successfully applied to the determination of copper in natural water and serum samples. PMID- 12769375 TI - A sequential injection method for the determination of Tween-80 in natural water samples using a fluorescence enhancement of the dye Eosin-B. AB - A simple sequential injection (SI) method is reported for the determination of the non-ionic surfactant polyoxyethylene sorbitan monoleate (Tween-80). The proposed method is based on a fluorescence enhancement of the probe 4',5'-dibromo 2',7'-dinitro fluorescein, disodium salt (Eosin B) in the presence of a surfactant. The procedure is optimized using the univariate method of optimization. The optimum operating conditions are as follows: reagent concentration, 40 ppm; pH, 4.5; aspirated volumes of sample and reagent, 120 microL and 100 microL, respectively. With the optimum conditions described, linear calibration curves were obtained from 10-200 ppm. The detection limit was 1.7 ppm and the maximum relative standard deviation of the method was 2.7% (n = 5). The fluorescence excitation and emission were fixed at lambda(ex) = 545 mn and lambda(em) = 585 nm. The method was successfully applied to the determination of (TW-80) in natural water samples. PMID- 12769376 TI - Determination of mercury(II) ion by electrochemical cold vapor generation atomic absorption spectrometry. AB - A technique for determination of mercury is described; it is based on electrolytic reduction of Hg(II) ion on a graphite cathode, the trapping of mercury vapor and its volatilization into a quartz tube aligned in the optical path of an atomic absorption spectrometer. The electrochemical cell consisted of a graphite cathode and an anode operating with constant direct current for the production of mercury atoms. A pre-activated graphite rod was used as the cathode material. The optimum conditions for electrochemical generation of mercury cold vapor (the electrolysis time and current, the flow rate, the type of electrode and electrolyte) were investigated. The characteristic electrochemical data with chemical cold vapor using NaBH4-acid were compared. The presence of cadmium(II), arsenic(III), antimony(III), selenium(IV), bismuth(III), silver(I), lead(II), lithium(I), sodium(I) and potassium(I) showed interference effects which were eliminated by suitable separation techniques. The calibration curve is linear over the range of 5-90 ng ml(-1) mercury(II). The detection limit is 2 ng ml(-1) of Hg(II) and the RSD is 2.5% (n = 10) for 40 ng ml(-1). The accuracy and recovery of the method were investigated by analyzing spiked tap water and river water. PMID- 12769377 TI - Spectrophotometric methods for the determination of anti-emetic drugs in bulk and in pharmaceutical preparations. AB - Four rapid, simple, reproducible and sensitive methods (A-D) for assaying domperidone (I) and metoclopramide (II) in a bulk sample and in dosage forms were investigated. The first and second methods, A and B, are based on the oxidation of I and/or II by Fe3+ in the presence of o-phenanthroline (o-phen) or bipyridyl (bipy). The formation of tris-complex upon reactions with Fe3+-o-phen and/or Fe3+ bipy mixture in an acetate buffer solution of the optimum pH-values was demonstrated. Methods C and D involve the addition of excess Ce4+ and the determination of unreacted oxidant by a decrease of the red color of chromotrope 2R (C2R) at a suitable lambda(max) of 528 nm for method C, or a decrease in the orange pink color of Rhodamine 6G (Rh6G) at a suitable lambda(max) value of 525 nm for method D. A regression analysis of Beer-Lambert plots showed a good correlation in the concentration range of 0.2-5.8 microg ml(-1). The apparent molar absorptivity, Sandell sensitivity, detection and quantification limits were calculated. For a more accurate analysis, the Ringbom optimum concentration ranges are 0.35-5.6 microg ml(-1). The developed methods were successfully applied to the determination of domperidone and metoclopramide in bulk and pharmaceutical preparations without any interference from common excipients. PMID- 12769378 TI - An algorithm for three-way data analysis that alternatively minimizes coupled vector (COV) resolution error and PARAFAC error. AB - A novel algorithm, alternatively minimizing coupled vector (COV) resolution error and PARAFAC error algorithm, is proposed in this paper. This algorithm can overcome the problem of slow convergence and is insensitive to the estimation of component number, such problems are unavoidable while using the traditional parallel factors analysis (PARAFAC) algorithm. In other words, this algorithm is capable of improving the computing speed and providing accurate resolutions provided that the number of factors used in the computation is no less than that of the actual underlying ones. The characteristic performances were demonstrated with a novel fluorescence data array. PMID- 12769379 TI - Monitoring the stability of an emulsion by high-frequency spectroscopy. AB - High-frequency spectroscopy has been applied to monitoring the stability of oil in-water emulsions. It was found that stable emulsions showed absorption peaks at around 550 MHz; further, they shifted from lower to higher frequencies with time. The rate of increase of the frequencies was dependent on the amount of emulsifier added. These results show that the stability of emulsions can be monitored by the amount of the frequency shift. PMID- 12769380 TI - Electrochemical and phosphorescent properties of new mixed-ligand Ir(II) complexes coordinated with both terpyridine and various bipyridine derivatives. AB - Seven useful mixed-ligand complexes in the form of [Ir(terpy)(L)Cl]2+ were prepared and their spectroscopic and electrochemical properties were investigated. The ligands used were terpy = 2,2':6',2''-terpyridine, L = 2,2' bipyridine, 4,4'-dimethyl-2,2'-bipyridine, 4,4'-diphenyl-2,2'-bipyridine, 1,10 phenanthroline, 5-phenyl-1,10-phenanthroline, 4,7-diphenyl-1,10-phenanthroline, 2,3-bis(2-pyridyl)pyrazine. Synthetic methods were developed by a sequential ligand-replacement which occurred in the reaction vessel using a microwave oven. All complexes showed that LUMOs are based on the pi-system contribution of the terpyridine ligand for [Ir(terpy)(bpy)Cl]2+, [Ir(terpy)(dmbpy)Cl]2+, [Ir(terpy)(dpbpy)Cl]2+, [Ir(terpy)(phen)Cl]2+, [Ir(terpy)(dpphen)Cl]2+ and [Ir(terpy)(phphen)Cl]2+. On the other hand, the LUMO in the [Ir(terpy)(bppz)Cl]2+ complex is localized on the pi-system of the bppz ligand, whereas the HOMOs in the iridium complexes are localized on the terpyridine ligand. It was found that Ir(terpy)(L)Cl emits in a fluid solution at room temperature. The ancillary ligands, such as terpy and bpy, have been explored to extend the lifetime of the triplet 3(pi-pi') excited states of Ir(III) terpyridine complexes. Ir(III) terpyridine units with an electron donor (dmbpy) or electron acceptor substituents (terpy, dpbpy, phphen, dpphen and bppz) are found to decrease the energy of the 3LC states for use as photosensitizer molecular components in supramolecular devices. The spectroscopic and electrochemical details are also reported herein. PMID- 12769381 TI - Electroanalytical properties of aldehyde biosensors with a hybrid-membrane composed of an enzyme film and a redox Os-polymer film. AB - Aldehyde biosensors were constructed by cross-linking formaldehyde dehydrogenase (FDH) or aldehyde dehydrogenase (ADH) and bovine serum albumin on the surface of a redox Os-polymer-coated electrode. The prepared aldehyde biosensors responded rapidly (within 30 s) to aldehydes without the addition of a soluble mediator, because the inner redox Os-polymer film effectively mediated the electron transfer from NADH generated enzymatically into the outer enzyme film to a glassy carbon electrode. An FDH/Os-polymer electrode responded linearly over the concentration range of 2 x 10(-6)-5 x 10(-4) M for formaldehyde, while an ADH/Os polymer electrode, though responding similarly to long chain aldehydes, such as propionaldehyde and butylaldehyde, responded linearly over the concentration range of 4 x 10(-6)-2 x 10(-4) M for acetaldehyde. PMID- 12769382 TI - Comparative study of the response of membrane electrodes based on calix[6]arene and calix[8]arene derivatives to organic ammonium ions. AB - p-tert-Butylcalix[8]arene-octaacetic acid octaethyl ester and calix[8]arene octaacetic acid octaethyl ester well recognized 2-phenylethylamine and phenylalanine methyl ester compared with the corresponding calix[6]arene derivatives. Moreover, the calix[8]arene derivatives, especially one having tert butyl groups, gave better selectivity against biologically active amines having a complicated structure, such as norephedrine. We considered the interaction between calixarenes and organic ammonium ions from the viewpoint of molecular symmetries. PMID- 12769383 TI - A nonaqueous potentiometric titration study of the dissociation of t-butyl methacrylate-methacrylic acid copolymers. AB - The dissociation of t-butyl methacrylate-methacrylic acid copolymers in dimethyl sulfoxide was analyzed by a nonaqueous potentiometric titration technique. The negative logarithm of the dissociation constant of the monomer unit of a methacrylic acid (MAA) monotonously increased with the increasing degree of dissociation corresponding to the titrant/MAA amount ratio, and was highly influenced by the copolymerization ratio. The results are discussed in terms of the suppression of the dissociation of MAA by a neighboring charged methacrylate anion unit. PMID- 12769384 TI - Use of an oxidation reaction for the quantitative determination of albendazole with Chloramine-T and acid dyes. AB - One titrimetric and two spectrophotometric procedures have been reported for the determination of albendazole and its tablets. Using titrimetry, the drug was titrated directly with Chloramine-T under acidic conditions using a Methyl Orange indicator. The spectrophotometric procedures involve treating the sample solution with a measured excess of Chloramine-T in an acid medium, followed by an estimation of unreacted Chloramine-T by reacting with a fixed amount of either Methyl Orange or Indigo Carmine dye solution and measuring the absorbance at 510 nm or 610 nm. The stoichiometric ratio, which forms the basis for the calculations in titrimetry as well as the range of the applicability, are reported. The Beer's law range and sensitivity values for spectrophotometric procedures are included. The methods were applied to the determination of albendazole in tablets with satisfactory results. PMID- 12769385 TI - Direct determination of benzamides in serum by column-switching high-performance liquid chromatography. AB - A column-switching high-performance liquid chromatographic method with fluorescence detection was developed for the simultaneous determination of four benzamide-type anti-psychotic drugs: sulpiride, tiapride, sultopride and metoclopramide in human serum. In this method, a TSKgel Super-ODS column was used as an analytical column, and a TSKgel G 2000SW was prepared as a pretreatment column. Under the optimized analytical conditions, four benzamide-type anti psychotic drugs were eluted within 18 min. The detection limits (S/N = 3) for sulpiride, tiapride, sultopride and metoclopramide are 1 ng/ml, 4 ng/ml, 2 ng/ml and 0.5 ng/ml, respectively. Finally, the method was applied to the determination of sulpiride in human serum samples obtained after a single oral dose of sulpiride. PMID- 12769386 TI - Simultaneous determination of trace elements in minute samples by electrothermal vaporization/low-pressure helium-ICP-MS. AB - Electrothermal vaporization with two filaments was studied for low-pressure helium-ICP-MS. A 10-microl volume of sample was dried and vaporized, and then introduced to the ICP. With this technique, four elements were simultaneously determined with the acceptable precisions (RSD ca. 10%). The detection limit based on 3sigma of background signals approached the fractional ppb range. The proposed method was successfully applied to the analysis of the certified reference material for human hair. PMID- 12769387 TI - Absorption spectrometry using a diffractive optical element constructed by gel electrophoresis. AB - A novel diffractive optical element was constructed by means of gel electrophoresis. Five capillaries were filled with a methylene blue solution and inserted into a slab gel. An electrical potential was then applied to the slab gel. Methylene Blue in the capillaries underwent electrophoretic migration, forming clear blue bands in the gel. The bands functioned as a transmission grating, and diffracted a laser beam. The intensity of the diffracted light increased with increasing the concentration of methylene blue. PMID- 12769388 TI - Crystal structure of [[mu-bis(salicylidene)-1,3 propanediaminato]copper(II)]dibromozinc(II). AB - The title compound is a double oxygen-bridged dimeric heteronuclear metal complex. The coordination around the Cu atom is distorted square-planar involving two O and two N atoms from the bis(salicylidene)-1,3-propanediamine ligand. The Zn atom in the molecule has a distorted tetrahedral coordination sphere consisting of the two O atoms of the ligand and the two Br atoms. The bridging plane between the metal atoms is not planar. PMID- 12769389 TI - Crystal structure of 1beta-hydroxy-beta-homopipitzolone (9-hydroxy-2,6,9 trimethyltricyclo[6,3,1,0(1,6)]dodeca-5,10,11,12-tetraone). AB - C15H18O5 is orthorhombic, P2(1)2(1)2(1). The unit-cell dimensions at 293 K are a = 6.526(3), b = 10.247(4), c = 21.008(13) A, V = 1404.8(12) A3, Dx = 1.316 g/cm3, and Z = 4. The R value is 0.044 for 1288 observed reflections. The A, B and C rings adopt chair, twist, and half-chair conformations. The absolute configuration is inferred from the C6 stereochemistry of (+)-cedrol. The crystal structure is stabilized by an O-H...O hydrogen bond and van der Waals forces. PMID- 12769390 TI - Crystal structure of a binuclear polymeric self-assembled lead(II) complex. AB - A polymeric self-assembled complex [[Pb(pydc)(pydc-H2)(H2O)2]2]n is prepared from the complexation of a novel pyridine containing self-assembling system, LH2, [pyda-H2]2+[pydc]2- (pyda = 2,6-pyridindiamine and pydc-H2 = 2,6 pyridinedicarboxilic acid) and lead(II) nitrate in 84% yield. The characterization was performed using X-ray crystallography. The crystal system is triclinic with space group P1 and two molecules per unit cell. The unit cell dimensions are a = 6.913(2) A, b = 10.687(4) A and c = 11.182(4) A with alpha = 92.805(6) degrees, beta = 101.821(6) degrees and gamma = 95.688(6) degrees. The final R value is 0.0373 for 4633 reflections measured. This compound is a nine coordinate binuclear complex with two metal fragments linked via the central four membered Pb2O2 ring. The crystal also contains a neutral [pydc-H2] molecule, that form hydrogen and coordination bonds that dominate the crystal packing, by forming layers of molecules. PMID- 12769391 TI - Crystal structure of zwitterionic trimethylammoniopropane sulfonate. AB - Crystal structure of zwitterionic trimethylammoniopropane sulfonate (Z1-Methyl) was determined by single-crystal X-ray diffractometry. Z1-Methyl crystallized in the monoclinic system, space group P2(1), with a = 6.3639(2), b = 10.9237(4), c = 8.1505(4) A, beta = 111.078(2) degrees, V = 528.69(4) A3, Z = 2. An inter molecular ionic association was observed in the crystal. The shortest distance between the methyl protons of the trimethylammonium group and the oxygens of the sulfonate group was 2.51(2) A. PMID- 12769392 TI - Crystal structure of dissymmetrically trihydrated N,N',N''-tribenzoylmelamine (TBM). AB - In the crystal structure of TBM-trihydrate three waters of crystallization were found to locate dissymmetrically. Thus, one of them forms a chelated ring with the neighboring C=O and N-H of TBM. Two other ones bind each other through hydrogen bonding and two sets of which form a cyclic dimer by hydrogen bonding (namely, water-tetramer). A C=O group binds by hydrogen bonding to each of the tetrameric water. The spatial volume occupied by water-tetramer appears to be sufficient for complexation with organic molecules. PMID- 12769393 TI - Validation of the Strengths and Stressors Tracking Device with a child welfare population. AB - The Strengths and Stressors Tracking Device (SSTD) is a rapid assessment measure of family well-being that assesses the particular strengths and needs of families at intake to help guide case planning and evaluate the effectiveness of treatment. The device assesses families from an ecological perspective in the domains of environmental conditions, social support, caregiver skills, and child well-being, and may be used at multiple points during treatment to assess change. SSTD has high internal consistency in all domains, distinguishes between physical abuse and neglect, and is sensitive to specific changes made by families across the duration of treatment. PMID- 12769395 TI - How parental drug use and drug treatment compliance relate to family reunification. AB - This study uses Cox regression to assess the relationships among parental drug use, drug treatment compliance, and reunification from substitute care. The study finds that drug treatment compliance is associated with faster reunification, even when accounting for ongoing drug use and three parenting measures. The findings are consistent with a conceptual framework suggesting that certain client actions, such as drug treatment compliance, may serve as markers that substantially affect client outcomes. PMID- 12769394 TI - Prenatal child abuse risk assessment: a preliminary validation study. AB - Workers need an efficient prenatal screener that can identify mothers at greatest risk of child abuse. Existing risk assessment methods are often invasive and difficult to administer. This study assessed child abuse risk in a sample of 49 expectant mothers using the Brigid Collins Risk Screener (BCRS). At three months postpartum, high-risk mothers scored significantly lower on the quality of infants' physical, social, and emotional environments than moderate or low-risk mothers. BCRS appears to offer a noninvasive, efficient approach to assessing risk of child abuse. PMID- 12769396 TI - Foster care and Medicaid managed care. AB - Children in the foster care system are often dependent on Medicaid for health care. These children, however, have more complex health care needs than the typical child receiving Medicaid. States are implementing Medicaid managed care programs as a way to control escalating costs while providing necessary services. This article reviews the issues surrounding delivery of managed health care services to children in foster care and describes several solutions. PMID- 12769397 TI - Nature of the beast. PMID- 12769398 TI - Supernumerary nipples: an overview. AB - Supernumerary nipples (SNs) are relatively common, minor congenital anomalies. SNs usually arise within the embryonic milk lines, but they have been known to form elsewhere. The prevalence of SNs varies, depending on the population. SNs are normally benign entities, but they are susceptible to hormonal changes and disease processes and may signify internal disease. PMID- 12769399 TI - Botanical briefs: Tulips--Tulipa species L. PMID- 12769400 TI - What is your diagnosis? Exostoses. PMID- 12769401 TI - Lasers, light, and acne. AB - Acne vulgaris is a highly prevalent disease with significant potential for physical and emotional scarring. Acne lesions have long been noted to improve after exposure to sunlight. This improvement may be secondary to activation of endogenous porphyrins produced by Propionibacterium acnes. Recently, several investigators have presented studies in which light of particular wavelengths has been used to treat acne vulgaris. In this article, we review the results of these studies as we look to the future of light-based acne treatment. PMID- 12769402 TI - Treatment of telangiectasia macularis eruptiva perstans with total skin electron beam radiation. AB - We describe the treatment of a 60-year old man with severely symptomatic telangiectasia macularis eruptiva perstans (TMEP) with a poor response to several standard therapeutic strategies. At that time, the patient underwent total skin electron beam (TSEB) radiation, based on the theory that by decreasing cutaneous mast cell infiltration, his pruritus would be relieved. He received a total dose of 4000 cGys given in 40 fractionated treatments. The patient had complete resolution of both his cutaneous lesions and pruritus, which has continued through one year of follow-up. PMID- 12769403 TI - Topical therapy for actinic keratoses, I: 5-Fluorouracil and imiquimod. AB - Actinic keratoses (AKs) are evolving, malignant cutaneous neoplasms. AKs can be treated with physical or destructive methods and with topical therapies. This article is the first in a 2-part series that will review current topical therapeutic options for AKs. Several topical treatment options offer some significant benefit for the alleviation of these lesions. Therapies include 5 fluorouracil, imiquimod, diclofenac, colchicine, and retinoids. The first part of this review will focus on topical 5-fluorouracil and imiquimod. PMID- 12769404 TI - Topical therapy for actinic keratoses, II: Diclofenac, colchicine, and retinoids. AB - Actinic keratoses (AKs) are evolving, malignant cutaneous neoplasms. AKs can be treated with physical or destructive methods and by topical therapies. This article is the second in a 2-part series of current topical therapeutic options for AKs and discusses topical diclofenac, colchicine, and retinoids. The first part focused on topical 5-fluorouracil and imiquimod. PMID- 12769405 TI - Actinic keratoses--surgical and physical therapeutic modalities. AB - Actinic keratoses (AKs) are the most common epithelial premalignant lesions seen by dermatologists today. The vast therapeutic armamentarium for treating AKs can be roughly divided into 2 categories: topical and surgical/physical modalities. It is important for clinicians to be familiarized with the various therapeutic options for treating AKs and to deliver individualized treatments. This article will review the surgical and physical modalities available for the treatment of AKs. PMID- 12769406 TI - Opposing views of 2 academies about the nature of solar keratosis. PMID- 12769407 TI - High dose cetirizine: a case report. AB - We report the case of a 46-year-old man who tolerated 50 mg per day of cetirizine for the treatment of chronic idiopathic urticaria. The patient denied any sedation or somnolence and had no difficulty performing routine daily functions including driving. He had tried other antihistamines, including fexofenadine, loratadine, and hydroxyzine without improvement. PMID- 12769408 TI - Teledermatology: an intraobserver diagnostic correlation study, part I. AB - Many studies have been published recently on the effectiveness of teledermatology as a diagnostic tool; however, much of the data comes from live 2-way video teleconferencing consultations and very little comes from more readily available "store and forward" consultations. Moreover, most published studies compare the diagnoses of 2 different dermatologists (interobserver comparison). Given the lack of data on baseline interdermatologist diagnostic variability, the interpretation of currently available diagnostic correlation data is somewhat difficult. The objective of this study is to measure the degree of diagnostic concordance between a dermatologist seeing a patient via a teledermatology consult system and the same dermatologist seeing the same patient face-to-face in a dermatology clinic at a tertiary medical center. A random sample of 404 patients was selected from patients who had routine appointments at our dermatology clinic. PMID- 12769409 TI - Pigmented basal cell carcinoma simulating melanoma in a burn scar. AB - Pigmented basal cell carcinoma (BCC) can masquerade as malignant melanoma, especially in dark-skinned patients. We present a rare case of pigmented BCC located in an old burn scar in a 74-year-old Hispanic woman, and we demonstrate the potential value of dermatoscopy as a diagnostic tool in cases of pigmented lesions. Although clinically simulating melanoma, findings from the dermatoscopic examination of the lesion revealed characteristics of pigmented BCC, which was confirmed by histologic analysis of the biopsy specimen. PMID- 12769410 TI - A sun protection survey of New England fishermen. AB - Skin cancer is the most common form of cancer in the United States, with more than 1 million cases of nonmelanoma skin cancer diagnosed annually and an estimated 53,600 cases of melanoma and 7400 deaths expected during the year 2002. Excess exposure to UV radiation has long been linked to both melanoma and nonmelanoma skin cancers. Fishing is a popular sport in the United States, and one with excessive sun exposure. Results of a survey of recreational fishermen conducted at the 23rd Annual Eastern Fishing & Outdoor Exposition found that sun protection attitudes and practices were poor or inadequate. I recommend that a tailored sun protection intervention program should take into account age, gender, and skin type, as well as the unique obstacles faced by fishermen. PMID- 12769411 TI - Treatment of molluscum contagiosum with the pulsed dye laser over a 28-month period. AB - Although benign, molluscum contagiosum causes cosmetic concern, infection, or transmission to close contacts. Treating patients with multiple lesions, especially children, may be difficult. Complications of treatment include infection, scarring, and limited posttreatment activity. The pulsed dye laser (PDL) has proven safe and effective for the treatment of many skin lesions, but little data exist for its role in the treatment of molluscum contagiosum. The purpose of this study is to describe the use, over a 28-month period, of the 585 nm PDL for the treatment of molluscum contagiosum. The benefits of this treatment are discussed. The charts of 43 patients receiving PDL treatment for molluscum contagiosum between November 1997 and March 2000 were reviewed. Number and location of lesions at initial presentation and on follow-up visits were recorded. Attempts were made to contact patients who were lost to follow-up. All of the approximately 1250 lesions treated resolved, and 35% of patients (n = 15) had no new lesions after 2 treatments. No complications were associated with the procedure. The PDL is a reasonable alternative to traditional treatment modalities for molluscum contagiosum. Benefits to the patient may include prompt response, fewer treatments, and minimal morbidity. PMID- 12769412 TI - Into the hidden world behind evidence-based medicine. PMID- 12769413 TI - Evidence-based medicine and quality of care. AB - In this paper we set out to examine the arguments for and against the claim that Evidence-Based Medicine (EBM) will improve the quality of care. In particular, we examine the following issues: 1. Are there hidden ethical assumptions in the methodology of EBM? 2. Is there a tension between the duty of care and EBM? 3. How can patient preferences be incorporated into quality guidelines and effectiveness studies? 4. Is there a tension between the quality of a particular intervention and overall quality of care? 5. Are certain branches of medicine and patient groups innately or prima facie disadvantaged by a shift to EBM? In addition we consider a case study in the ethics of EBM, on a clinical trial concerning the collection of umbilical cord blood in utero and ex utero, during or after labour in childbirth. PMID- 12769414 TI - Evidence-based medicine as an instrument for rational health policy. AB - This article tries to present a broad view on the values and ethical issues that are at stake in efforts to rationalize health policy on the basis of economic evaluations (like cost-effectiveness analysis) and randomly controlled clinical trials. Though such a rationalization is generally seen as an objective and 'value free' process, moral values often play a hidden role, not only in the production of 'evidence', but also in the way this evidence is used in policy making. For example, the definition of effectiveness of medical treatment or health care service is heavily dependent on dominant individual or social views about the goals of the particular treatment or service. There is also a concern that a reliance on EBM in health policy will occur at the expense of widely shared social values like equity and solidarity. Moreover, there is a concern that when economic considerations and rational procedures become more influential, various 'outside' groups third parties like insurance companies and policy makers will get a stronger influence on medical practice which may lead to a change in the patient-provider relationship. The authors conclude that social values and patient preference should be explicitly addressed when health policy making is based on economic and other scientific evidence. PMID- 12769415 TI - Is there a tension between doctors' duty of care and evidence-based medicine? AB - The interaction between evidence-based medicine and doctors' duty of care to patients is complex. One the one hand, there is surely an obligation to take account of the best available evidence when offering health care to patients. On the other hand, it is equally important to be aware of important shortcomings in the processes and practices of evidence-based medicine. There are tensions between the population focus of evidence-based medicine and the duties that doctors have to individual patients. Implementing evidence-based medicine may have unpredictable consequences upon the overall quality of health care. Patients may have a range of reasons for preferring one form of treatment over another, not all of which are captured by current formulations of evidence. This paper examines these issues, using relevant examples from evidence-based medicine. PMID- 12769416 TI - Managing chronic disease: evidence-based medicine or patient centred medicine? AB - Chronic diseases are recognized as a leading cause of mortality, morbidity, health care utilization and cost. A constant tailoring of care to the actual needs of individual patients, complexity and long duration are the distinguishing features of chronic disease management. Given the rapid development and high use of services providing complex management, the number of controlled clinical trials in this field is limited. The information from the few available controlled clinical trials may be difficult to interpret, mainly due to a large variety in the interventions being studied, differences in 'control treatments' and a confined set of outcome measures that are used. The ethical issue with this observation is, that in the absence of randomised clinical trial information on clinical effectiveness and in consequence of the lack of additional data that are crucial for therapeutic decisions in the process of caring, specific patient groups, such as patients with chronic diseases, may become disadvantaged. The scarcity and incompleteness of controlled trial information can partly be explained by difficulties in conducting this type of research in the field of chronic disease management. To avoid that patients with chronic diseases become disadvantaged, the use of alternative designs such as observational studies to evaluate chronic disease management must be accepted and supported. Moreover, in chronic disease management the process of caring needs to emphasized and appraised appropriately. For that purpose, new measurement methods, focussing on concepts of caring that are not included in the majority of current clinical trials, need to be developed. PMID- 12769417 TI - Evidence-based medicine: excessive attraction to efficiency and certainty? AB - Advocates of EBM deserve much credit for their efforts to increase the use of scientific evidence and economic evaluation in medical decision making. But EBM advocates' rigid requirements of certainty in the estimation of intervention effects may run counter to society's interest in maximising the expected benefits from resource use in health care. Also, their dedication to efficiency may lead some to overlook societal concerns for fairness in resource allocation. PMID- 12769418 TI - The role of evidence in health policy making: a normative perspective. AB - Assessment of evidence is becoming a central part of health policy decisions--not least in limit setting decisions. Limit-setting decisions can be defined as the withholding of potentially beneficial health care. This article seeks to explore the value choices related to the use of evidence in limit-setting decisions at the political level. To better specify the important but restricted role of evidence in such decisions, the value choices of relevance are discussed explicitly. Four criteria are often considered when setting limits: 1. The severity of disease if untreated or treated by standard care 2. The effectiveness of the new technology 3. The cost-effectiveness of the new technology 4. The quality of evidence on (1)-(3) The production and assessment of evidence is important for each criterion, but several points are identified where the practice of evidence-based medicine could be further developed to capture a broader spectrum of ethical and political concerns that such decisions naturally evoke among citizens. PMID- 12769419 TI - Evidence-based medicine and power shifts in health care systems. AB - It is important and urgent to question the relationship between evidence-based medicine and power shifts in health care systems. Although definitions of EBM are phrased as a scientific approach to medicine, EBM is a normative concept: it aims to improve medicine and health care. Both proponents and opponents use a normative concept. More particularly, they provide particular views on positions, responsibilities, possibilities, norms and relationships between professionals, patient groups, governments and other parties in health care and society. From this perspective, we want to analyse the role of EBM in modern western societies. By using citizenship theory, we will argue that the role of EBM is not fixed but depends on the relation between state and society. We will first analyse the fundamental change in western societies during the past decades, from modern to post-modern societies. Then, we will elaborate a fourfold model of possible relationships between state and society, and discuss the issue of how EBM may fit in, by giving some examples of the practice of EBM in different European countries. On this basis, we conclude to consider EBM as a public forum where proponents and opponents of EBM discuss diverse and possibly conflicting ways of changing medicine, health care, and health policy. This requires the incorporation of the perspective of citizens and their social networks, professionals with practical and tacit knowledge, and diverse public views on what is regarded as 'a good life'. Inasmuch as EBM is expected to be practically relevant, it ought to be tied to rather than separated from the normative world of emancipated patients and diverse health care practices. Proponents and opponents of EBM should be prepared to defend the normative claims and power effects that are inherently tied to any presentation of evidence. PMID- 12769420 TI - Legalising science. AB - The legal view of science has changed through time, moving from a positivist and noncritical position of law towards science to a critical view of science- providing the potential for more objective knowledge, but value-laden as well- and of the role of society. This paper explores some judicial cases that illustrate these attitudes, suggesting that reference to science (particularly to EBM) can be rigorously and equitably made when it serves the cause of transparency and democratisation both in science and in law. PMID- 12769421 TI - Understanding the introvert preference. PMID- 12769422 TI - Teaching students to apply nursing theories and models: trying something new. PMID- 12769423 TI - Grade disputes: considerations for nursing faculty. AB - Nursing faculty take grading of assignments seriously. Faculty realize students are sensitive about their grade point averages and that students must maintain certain grades for progression and the retention of scholarships. Faculty further recognize that the grading of clinical performance is complex and subjective by nature. Some faculty are reluctant to assign a failing grade for poor clinical performance for fear of litigation. In this article, the authors discuss several landmark cases that have come before the courts, which have set precedence for grade disputes. The courts overwhelmingly have supported faculty decisions regarding grade assignment, as long as the grades were not arbitrary or capricious. Nursing instructors should not be fearful of failing a student solely on the basis of poor clinical performance. However, faculty must be prepared to explain how grades are assessed related to the program and course objectives. PMID- 12769424 TI - Patterns of peer tutoring in nursing. AB - Peer tutoring in higher education is an effective strategy for promoting academic gains. Within nursing, peer tutoring has been used in the clinical setting, but little information is available regarding its use across the nursing curriculum. A peer tutoring program was created at a regional Appalachian university to meet the needs of students with poor academic backgrounds and multiple risk factors for failure. As the program naturally evolved, students moved beyond the time honored one-on-one model. Many tutoring patterns developed including dyad, small group, large group, skill based, assignment based, and question based. Qualitative evaluation data from the program revealed that each pattern required different tutor skills, involved varied tutor-tutoree relationships, focused on different outcomes, and had certain advantages and disadvantages. All tutoring patterns contributed to improved academic skills and performance. PMID- 12769425 TI - Using gaming to help nursing students understand ethics. AB - The authors developed an ethics game that uses specially designed ethical situations for students to consider. Two students argue a course of action based on the scenario and defend that action using content discussed in class. Substantive issues include decision-making models, values as they pertain to the situation, professional responsibilities, ethical principles, social expectations, and legal requirements. Points are awarded based on how compelling each argument is. All students have an opportunity to participate. The benefits of using the game are that students gain confidence in their ability to defend an ethical decision, are able to see ethical situations from more than one perspective, and have an opportunity to clarify values. In addition, ethical principles and decision-making models are brought to life in a fun way. Difficulties involved in using the game include class size and limited time between the students learning course content and using it in the game. PMID- 12769426 TI - Establishing student competency in qualitative research: can undergraduate nursing students perform qualitative data analysis? AB - Many undergraduate nursing programs require some level of research competency of their graduates. However, the competency level may vary depending on program goals and often is focused on quantitative methodologies. The purpose of this research was to determine nursing students' ability to perform qualitative data analysis. Twenty-eight undergraduate students enrolled in a data analysis course in the junior year of their baccalaureate program participated. After selecting the research question, students collected interview data from each other and posted it using a Web-based program. The students and instructor then performed a content analysis. After reviewing the instructor's analysis, which revealed three core themes, students' written analyses were read to determine their congruence with the instructor-generated themes. There was 89% agreement on the first theme, 96% agreement on the second theme, and 61% agreement on the third theme. Because of the high agreement rates between the instructor and student analyses, the author concluded that undergraduate nursing students, when provided with appropriate instruction, are capable of performing qualitative data analysis. PMID- 12769427 TI - Reading statistics in nursing research: a self-study CD-ROM module. AB - A statistics CD-ROM tutorial program was developed to replace a classroom course with several self-study modules. For the CD-ROM delivery method, students were asked to complete a Likert-type questionnaire with response options similar to those used for end-of-course evaluations at the nursing school. Ratings ranged from 1 to 5, with 1 equaling outstanding and 5 equaling completely unsatisfactory. Ratings for the CD-ROM were compared with those of a Web-based course taught the prior year. Mean satisfaction ratings of Web-based delivery of content was low due to technical problems with the Internet courseware used. Overall satisfaction with the CD-ROM for students who used all the components was improved substantially, compared to the Web-based delivery method. PMID- 12769428 TI - Using a linked learning activity to foster nursing students' professional growth. AB - Linked learning assignments employ a learner-centered approach that views differences among students as educationally enriching, rather than problematic. This article describes a linked learning activity that systematically incorporated differences between first-year and fourth-year nursing students to enrich learning outcomes related to professional growth and the image of nursing. The process of developing, implementing, and evaluating this professional growth linked learning activity is described. Thematic analysis of students' and faculty's reflective writing about their experience of the learning activity generated three themes--experiencing collegiality, sharing insider perspectives, and re-envisioning nursing. Suggestions for using linked learning activities to actively engage the diversity present in nursing undergraduate programs are offered. The value of facilitating and providing supportive environments for dialogue among students at various stages of development to promote positive socialization in nursing is discussed. PMID- 12769429 TI - Initiation into qualitative data analysis. AB - Qualitative data analysis may be daunting to nursing graduate students and other novice qualitative researchers. This article describes a qualitative research project required in a graduate nursing research course that was desired to guide students through their first qualitative data analysis. Common pitfalls novice researchers may encounter, such as data shuffling, premature closure, and overly delayed closure, also are discussed. The project reported in this article not only decreased students' anxiety concerning qualitative data analysis but also helped students gain respect for the amount of time, effort, and creativity this process entails. PMID- 12769430 TI - Dynamical diseases of brain systems: different routes to epileptic seizures. AB - In this overview, we consider epilepsies as dynamical diseases of brain systems since they are manifestations of the property of neuronal networks to display multistable dynamics. To illustrate this concept we may assume that at least two states of the epileptic brain are possible: the interictal state characterized by a normal, apparently random, steady-state electroencephalography (EEG) ongoing activity, and the ictal state, that is characterized by paroxysmal occurrence of synchronous oscillations and is generally called, in neurology, a seizure. The transition between these two states can either occur: 1) as a continuous sequence of phases, like in some cases of mesial temporal lobe epilepsy (MTLE); or 2) as a sudden leap, like in most cases of absence seizures. In the mathematical terminology of nonlinear systems, we can say that in the first case the system's attractor gradually deforms from an interictal to an ictal attractor. The causes for such a deformation can be either endogenous or external. In this type of ictal transition, the seizure possibly may be anticipated in its early, preclinical phases. In the second case, where a sharp critical transition takes place, we can assume that the system has at least two simultaneous interictal and ictal attractors all the time. To which attractor the trajectories converge, depends on the initial conditions and the system's parameters. An essential question in this scenario is how the transition between the normal ongoing and the seizure activity takes place. Such a transition can occur either due to the influence of external or endogenous factors or due to a random perturbation and, thus, it will be unpredictable. These dynamical changes may not be detectable from the analysis of the ongoing EEG, but they may be observable only by measuring the system's response to externally administered stimuli. In the special cases of reflex epilepsy, the leap between the normal ongoing attractor and the ictal attractor is caused by a well-defined external perturbation. Examples from these different scenarios are presented and discussed. PMID- 12769431 TI - Epileptic seizure prediction and control. AB - Epileptic seizures are manifestations of epilepsy, a serious brain dynamical disorder second only to strokes. Of the world's approximately 50 million people with epilepsy, fully 1/3 have seizures that are not controlled by anti-convulsant medication. The field of seizure prediction, in which engineering technologies are used to decode brain signals and search for precursors of impending epileptic seizures, holds great promise to elucidate the dynamical mechanisms underlying the disorder, as well as to enable implantable devices to intervene in time to treat epilepsy. There is currently an explosion of interest in this field in academic centers and medical industry with clinical trials underway to test potential prediction and intervention methodology and devices for Food and Drug Administration (FDA) approval. This invited paper presents an overview of the application of signal processing methodologies based upon the theory of nonlinear dynamics to the problem of seizure prediction. Broader application of these developments to a variety of systems requiring monitoring, forecasting and control is a natural outgrowth of this field. PMID- 12769432 TI - Manipulating epileptiform bursting in the rat hippocampus using chaos control and adaptive techniques. AB - Epilepsy is a relatively common disease, afflicting 1%-2% of the population, yet many epileptic patients are not sufficiently helped by current pharmacological therapies. Recent reports have suggested that chaos control techniques may be useful for electrically manipulating epileptiform bursting behavior in vitro and could possibly lead to an alternative method for preventing seizures. We implemented chaos control of spontaneous bursting in the rat hippocampal slice using robust control techniques: stable manifold placement (SMP) and an adaptive tracking (AT) algorithm designed to overcome nonstationarity. We examined the effect of several factors, including control radius size and synaptic plasticity, on control efficacy. AT improved control efficacy over basic SMP control, but relatively frequent stimulation was still necessary and very tight control was only achieved for brief stretches. A novel technique was developed for validating period-1 orbit detection in noisy systems by forcing the system directly onto the period-1 orbit. This forcing analysis suggested that period-1 orbits were indeed present but that control would be difficult because of high noise levels and nonstationarity. Noise might actually be lower in vivo, where regulatory inputs to the hippocampus are still intact. Thus, it may still be feasible to use chaos control algorithms for preventing epileptic seizures. PMID- 12769433 TI - Spatio-temporal dynamics prior to neocortical seizures: amplitude versus phase couplings. AB - The mechanisms underlying the transition of brain activity toward epileptic seizures remain unclear. Based on nonlinear analysis of both intracranial and scalp electroencephalographic (EEG) recordings, different research groups have recently reported dynamical smooth changes in epileptic brain activity several minutes before seizure onset. Such preictal states have been detected in populations of patients with mesial temporal lobe epilepsy (MTLE) and, more recently, with different neocortical partial epilepsies (NPEs). In this paper, we are particularly interested in the spatio-temporal organization of epileptogenic networks prior to seizures in neocortical epilepsies. For this, we characterize the network of two patients with NPE by means of two nonlinear measures of interdependencies. Since the synchronization of neuronal activity is an essential feature of the generation and propagation of epileptic activity, we have analyzed changes in phase synchrony between EEG time series. In order to compare the phase and amplitude dynamics, we have also studied the degree of association between pairs of signals by means of a nonlinear correlation coefficient. Recent findings have suggested changes prior to seizures in a wideband frequency range. Instead, for the examples of this study, we report a significant decrease of synchrony in the focal area several minutes before seizures (>>30 min in both patients) in the frequency band of 10-25 Hz mainly. Furthermore, the spatio-temporal organization of this preictal activity seems to be specifically related to this frequency band. Measures of both amplitude and phase coupling yielded similar results in narrow-band analysis. These results may open new perspectives on the mechanisms of seizure emergence as well as the organization of neocortical epileptogenic networks. The possibility of forecasting the onset of seizures has important implications for a better understanding, diagnosis and a potential treatment of the epilepsy. PMID- 12769434 TI - Channel-consistent forewarning of epileptic events from scalp EEG. AB - Phase-space dissimilarity measures (PSDM) have been recently proposed to provide forewarning of impending epileptic events from scalp electroencephalographic (EEG) for eventual ambulatory settings. Despite high noise in scalp EEG, PSDM yield consistently superior performance over traditional nonlinear indicators, such as Kolmogorov entropy, Lyapunov exponents, and correlation dimension. However, blind application of PSDM may result in channel inconsistency, whereby multiple datasets from the same patient yield conflicting forewarning indications in the same channel. This paper presents a first attempt to solve this problem. PMID- 12769435 TI - Efficient estimation of a time-varying dimension parameter and its application to EEG analysis. AB - This paper considers the problem of estimating the dimension of nonstationary electroencephalogram (EEG) signals and describes the implementation of an efficient algorithm to calculate a time-varying dimension estimate. The algorithm allows the practical calculation of a dimension estimate and its statistical significance over large data sets with a high temporal resolution. The method is applied to EEG recordings from patients with temporal lobe epilepsy and in one case the results of the analysis are compared with those obtained from an existing method of computing the correlation density. PMID- 12769436 TI - Epileptic seizure prediction using hybrid feature selection over multiple intracranial EEG electrode contacts: a report of four patients. AB - Epileptic seizure prediction has steadily evolved from its conception in the 1970s, to proof-of-principle experiments in the late 1980s and 1990s, to its current place as an area of vigorous, clinical and laboratory investigation. As a step toward practical implementation of this technology in humans, we present an individualized method for selecting electroencephalogram (EEG) features and electrode locations for seizure prediction focused on precursors that occur within ten minutes of electrographic seizure onset. This method applies an intelligent genetic search process to EEG signals simultaneously collected from multiple intracranial electrode contacts and multiple quantitative features derived from these signals. The algorithm is trained on a series of baseline and preseizure records and then validated on other, previously unseen data using split sample validation techniques. The performance of this method is demonstrated on multiday recordings obtained from four patients implanted with intracranial electrodes during evaluation for epilepsy surgery. An average probability of prediction (or block sensitivity) of 62.5% was achieved in this group, with an average block false positive (FP) rate of 0.2775 FP predictions/h, corresponding to 90.47% specificity. These findings are presented as an example of a method for training, testing and validating a seizure prediction system on data from individual patients. Given the heterogeneity of epilepsy, it is likely that methods of this type will be required to configure intelligent devices for treating epilepsy to each individual's neurophysiology prior to clinical deployment. PMID- 12769437 TI - Adaptive epileptic seizure prediction system. AB - Current epileptic seizure "prediction" algorithms are generally based on the knowledge of seizure occurring time and analyze the electroencephalogram (EEG) recordings retrospectively. It is then obvious that, although these analyses provide evidence of brain activity changes prior to epileptic seizures, they cannot be applied to develop implantable devices for diagnostic and therapeutic purposes. In this paper, we describe an adaptive procedure to prospectively analyze continuous, long-term EEG recordings when only the occurring time of the first seizure is known. The algorithm is based on the convergence and divergence of short-term maximum Lyapunov exponents (STLmax) among critical electrode sites selected adaptively. A warning of an impending seizure is then issued. Global optimization techniques are applied for selecting the critical groups of electrode sites. The adaptive seizure prediction algorithm (ASPA) was tested in continuous 0.76 to 5.84 days intracranial EEG recordings from a group of five patients with refractory temporal lobe epilepsy. A fixed parameter setting applied to all cases predicted 82% of seizures with a false prediction rate of 0.16/h. Seizure warnings occurred an average of 71.7 min before ictal onset. Similar results were produced by dividing the available EEG recordings into half training and testing portions. Optimizing the parameters for individual patients improved sensitivity (84% overall) and reduced false prediction rate (0.12/h overall). These results indicate that ASPA can be applied to implantable devices for diagnostic and therapeutic purposes. PMID- 12769438 TI - Comparison of predictability of epileptic seizures by a linear and a nonlinear method. AB - The performance of traditional linear (variance based) methods for the identification and prediction of epileptic seizures are contrasted with "modern" methods from nonlinear time series analysis. We note several flaws of design in demonstrations claiming to establish the efficacy of nonlinear techniques; in particular, we examine published evidence for precursor identification. We perform null hypothesis tests using relevant surrogate data to demonstrate that decreases in the correlation density prior to and during seizure may simply reflect increases in the variance. PMID- 12769439 TI - Discerning nonstationarity from nonlinearity in seizure-free and preseizure EEG recordings from epilepsy patients. AB - A number of recent studies indicate that nonlinear electroencephalogram (EEG) analyses allow to define a state predictive of an impending epileptic seizure. In this paper, we combine a method for detecting nonlinear determinism with a novel test for stationarity to characterize EEG recordings from both the seizure-free interval and the preseizure phase. We discuss differences between these periods, particularly an increased occurrence of stationary, nonlinear segments prior to seizures. These differences seem most prominent for recording sites within the seizure-generating area and for EEG segments less than one minute's length. PMID- 12769440 TI - Prediction of PTZ-induced seizures using wavelet-based residual entropy of cortical and subcortical field potentials. AB - Our proposed algorithm for seizure prediction is based on the principle that seizure build-up is always preceded by constantly changing bursting levels. We use a novel measure of residual subband wavelet entropy (RSWE) to directly estimate the entropy of bursts, which is otherwise obscured by the ongoing background activity. Our results are obtained using a slow infusion anesthetized pentylenetetrazol (PTZ) rat model in which we record field potentials (FPs) from frontal cortex and two thalamic areas (anterior and posterior nuclei). In each frequency band, except for the theta-delta frequency bands, we observed a significant build-up of RSWE from the preictal period to the first ictal event (p < or = 0.05) in cortex. Significant differences were observed between cortical and thalamic RSWE (p < or = 0.05) subsequent to seizure development. A key observation is the twofold increase in mean cortical RSWE from the preictal to interictal period. Exploiting this increase, we develop a slope change detector to discern early acceleration of entropy and predict the approaching seizure. We use multiple observations through sequential detection of slope changes to enhance the sensitivity of our prediction. Using the proposed method applied to a cohort of four rats subjected to PTZ infusion, we were able to predict the first seizure episode 28 min prior to its occurrence. PMID- 12769441 TI - Role of NK1.1+ and AsGm-1+ cells in oral immunoregulation of experimental colitis. AB - NK1.1 and AsGm-1 expressing cells play a role in immunomodulation. Our purpose was to determine the role of NK1.1+ and AsGm-1+ expressing cells in the inflammatory/tolerance paradigm in experimental colitis. Oral tolerance towards colitis-extracted proteins had previously been shown to alleviate experimental colitis. Colitis was induced in C57/B6 mice by intracolonic instillation of trinitrobenzenesulfonic acid (TNBS). Oral tolerance was induced via five oral doses of proteins extracted from TNBS-colitis colonic wall. Clinical, macroscopic, and microscopic scores were used for colitis assessment. To evaluate the putative role of AsGm-1 in tolerance induction, depletion of AsGm-1 expressing cells was performed. To evaluate the mechanism of tolerance induction, liver-associated NKT lymphocytes were harvested 14 days following tolerance induction, and cultured with concanavalin A (con A) and colitis-extracted proteins. T cell subsets were measured by flow cytometry. Cytokine expression was measured by intracellular staining and enzyme-linked immunosorbent assay (ELISA). Orally tolerized mice exhibited significant alleviation of the clinical, macroscopic, and microscopic parameters of colitis, with increased CD4+ILA+/CD4+IFNgamma+ lymphocyte ratio, increased IL-4, and decreased IFNgamma and IL-12 serum levels. In contrast, orally fed mice that were AsGm-1 depleted showed evidence of severe colitis. These mice exhibited significant decreased CD4 +IL4+/CD4+IFNgamma+ ratios, and an increase in IFNgamma and IL-12, with decreased IL-4 levels. NKT cells harvested from tolerized mice secreted high levels of antiinflammatory cytokines. In contrast, in nontolerized mice, NKT cells mainly secreted proinflammatory cytokines. In a tolerized environment, both NK1.1 and AsGm-1 expressing cells are essential for disease alleviation. In contrast, in a nontolerized environment, AsGm-1 expressing cells support an antiinflammatory immune paradigm, while NKT lymphocytes support a proinflammatory shift. PMID- 12769442 TI - Lack of interleukin-10 leads to intestinal inflammation, independent of the time at which luminal microbial colonization occurs. AB - Previous studies have demonstrated that the resident bacteria harbored by interleukin (IL)-10 gene-deficient mice initiate an enterocolitis in the neonatal period. The associated intestinal injury is characterized by an increase in the secretion of interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha, and by a systemic response to endogenous bacterial antigens, supporting the hypothesis that a lack of tolerance may be the initiating cause. Whether bacterial initiation of this enterocolitis would occur in the adult intestine or whether it is only seen in the developing neonatal intestine was not known. Adult (9 weeks of age), axenic, luminally sterile IL-10 gene-deficient mice, which do not spontaneously develop enterocolitis, were inoculated with intestinal microbial flora. These mice rapidly developed intestinal injury and demonstrated elevated levels of IFN-gamma in cecal and colonic tissue. This response precedes a systemic spleen cell response to stimulation by bacterial antigens. Similarly, axenic, IL-10 gene-deficient mice exposed to microflora as neonates experience a comparable intestinal injury and IFN-gamma release before the appearance of IFN gamma-producing cells in the spleen. Microbial colonization in control mice leads to systemic IL-10 production, but not systemic IFN-gamma production, suggesting that an IL-10-mediated suppression regulates the response in normal control mice but is absent in IL-10 gene-deficient mice. Our results suggest that the point at which intestinal microbial colonization occurs does not significantly influence the severity or specificity of the inflammatory response in IL-10 gene-deficient mice. The lack of tolerance to bacterial antigens appears to result from the absence of IL-10 during bacterial exposure. PMID- 12769443 TI - Treatment of perianal fistulizing Crohn's disease with infliximab alone or as an adjunct to exam under anesthesia with seton placement. AB - Perianal fistulas occur in approximately 30% of patients with Crohn's disease (CD). Infliximab, a chimeric monoclonal antibody targeting human tumor necrosis factor alpha (TNF), is approved for the treatment of fistulizing CD. Although the initial response to infliximab is dramatic, the median duration of fistula closure is approximately 3 months, and repeated infusions are often required. An exam under anesthesia (EUA) by a surgeon allows for complete inspection of the fistula as well as incision and drainage of an abscess and placement of a seton. Our aim was to compare the rate of perianal fistula healing, relapse rate, and time to relapse in patients with fistulizing CD treated with infliximab alone or as an adjunct to surgical EUA with seton placement. Thirty-two consecutive patients with perianal fistulizing CD who completed at least 3 infusions with infliximab (5 mg/kg at 0, 2, 6 weeks) between October 1999 and October 2001 were analyzed. All patients had at least 3 months of follow-up after the third dose of infliximab. Response was defined as complete closure and cessation of drainage from the fistula. Patients with CD and perianal fistulas who had an EUA prior to infliximab infusions had a better initial response (100% vs. 82.6%, p = 0.014), lower recurrence rate (44% vs. 79%, p = 0.001), and longer time to recurrence (13.5 months vs. 3.6 months, p = 0.0001) compared with patients receiving infliximab alone. In conclusion, patients with fistulizing CD treated with infliximab are more likely to maintain fistula closure if treatment is preceded by EUA and seton placement. PMID- 12769444 TI - Pulmonary manifestations of inflammatory bowel disease. AB - Extraintestinal manifestations of both Crohn's disease and ulcerative colitis (UC) have been well described, although pulmonary findings are often overlooked. We summarize the experience of more than 400 cases of pulmonary manifestations of inflammatory bowel disease (IBD). These manifestations will be categorized by disease mechanism into drug-induced disease, anatomic disease, over-lap syndromes, autoimmune disease, physiologic consequences of IBD, pulmonary function test abnormalities, and nonspecific lung disease. We intend to provide the clinician with a practical working update on the spectrum of pulmonary dysfunction associated with IBD. PMID- 12769445 TI - Increasing fecal butyrate in ulcerative colitis patients by diet: controlled pilot study. AB - Topical butyrate has been shown to be effective in the treatment of ulcerative colitis (UC). Butyrate is derived from colonic fermentation of dietary fiber, and our aim was to study whether UC patients could safely increase the fecal butyrate level by dietary means. We enrolled 22 patients with quiescent UC (mean age, 44 years; 45% women; median time from last relapse, 1 year) in a controlled pilot trial lasting 3 months. The patients were instructed to add 60 g oat bran (corresponding to 20 g dietary fiber) to the daily diet, mainly as bread slices. Fecal short-chain fatty acids (SCFAs) including butyrate, disease activity, and gastrointestinal symptoms were recorded every 4 weeks. During the oat bran intervention the fecal butyrate concentration increased by 36% at 4 weeks (from 11 +/- 2 (mean +/- SEM) to 15 +/- 2 micromol/g feces) (p < 0.01). The mean butyrate concentration over the entire test period was 14 +/- 1 micromol/g feces (p < 0.05). Remaining fecal SCFA levels were unchanged. No patient showed signs of colitis relapse. Unlike controls, the patients showed no increase in gastrointestinal complaints during the trial. Yet patients reporting abdominal pain and reflux complaints at entry showed significant improvement at 12 weeks that returned to baseline 3 months later. This pilot study shows that patients with quiescent UC can safely take a diet rich in oat bran specifically to increase the fecal butyrate level. This may have clinical implications and warrants studies of the long-term benefits of using oat bran in the maintenance therapy in UC. PMID- 12769446 TI - Case report: magnetic resonance imaging in the diagnosis of epidural abscess complicating perirectal fistulizing Crohn's disease. AB - Epidural abscess is a rare complication of fistulizing Crohn's disease (CD), potentially appearing as neurologic symptoms or back and leg pain. We report a case of a large epidural abscess resulting from uncontrolled fistulizing CD, which was rapidly defined using gadolinium-enhanced magnetic resonance imaging (MRI). Whenever caudal neurologic symptoms, back pain, and fever arise in CD patients, diagnostic MRI of the pelvis in addition to conventional computerized tomography should be considered to identify perirectal fistulization to the spine. PMID- 12769447 TI - Gastroduodenal Crohn's disease: medical management. PMID- 12769448 TI - Severe gastroduodenal Crohn's disease: surgical treatment. PMID- 12769449 TI - Keeping the M in mind: recombinant human GMCSF and Crohn's disease. PMID- 12769450 TI - FGF-20: FGFs--they keep growing and growing and growing! PMID- 12769452 TI - Another new look at the small bowel: feasibility of three-dimensional magnetic resonance imaging to evaluate Crohn's disease. PMID- 12769451 TI - A map to the diagnosis of Crohn's disease. PMID- 12769453 TI - Competition asymmetry with taxon divergence. AB - Most organisms experience competition for resources, probably most of the time. As the structure and requirements of closely related species are generally liable to be more similar than in distantly linked species, Darwin suggested that the potential for competition was greater in the former. Since that time, studies have concentrated on interactions of either conspecifics or congeneric species. Shared critical resources, which organisms compete for, are generally mates, food and space (for access to the former). Whilst mates are valued only within species, in that the definition of a species requires it so, both food and space have the potential to be shared by very different organisms. It is now clear that vertebrates may compete with remotely related species: e.g. with squid for krill and with insects for nectar or seeds. Diamond suggested that (i) mutual aggression, (ii) displacement and (iii) evolutionary change in morphology would be increasingly asymmetric with competitor dissimilarity. Thus, with increasing taxonomic distance between two competitors (A and B), increasing aggression is exhibited between them and, increasingly, one consistently displaces the other. Here, Darwin's suggestion and Diamond's first two theories are tested across a taxonomic spectrum for the first time to the best of the author's knowledge. The proportion of spatial competitors in two different marine invertebrate groups demonstrating mutual aggression and displacement increases with taxon divergence (Nei's genetic identity). Congenerics were twice as likely to fight as conspecifics, and confamilial competitors were three times as likely to fight as conspecifics. This relationship seems robust to taxonomic and environmental variability. Competitors do not need to be as distant as birds and bees for complete asymmetry, a different family seems sufficient. PMID- 12769454 TI - Effects of Microphallus papillorobustus (Platyhelminthes: Trematoda) on serotonergic immunoreactivity and neuronal architecture in the brain of Gammarus insensibilis (Crustacea: Amphipoda). AB - The larval flatworm Microphallus papillorobustus encysts in the protocerebrum of its intermediate host, Gammarus insensibilis, and changes the gammarid's responses to mechanical and photic stimuli. The resulting aberrant escape behaviour renders infected gammarids more susceptible to predation by birds, the definitive hosts of the parasite. We used immunocytochemical methods to explore the mechanisms underlying these subtle behavioural modifications. Whole mounts of gammarid brains were labelled with fluorescent anti-serotonin and anti-synapsin antibodies and viewed using confocal microscopy. Two types of change were observed in infected brains: the intensity of the serotonergic label was altered in specific regions of the brain, and the architecture of some serotonergic tracts and neurons was affected. A morphometric analysis of the distribution of the label showed that serotonergic immunoreactivity was decreased significantly (by 62%) in the optic neuropils, but not in the olfactory lobes, in the presence of the parasite. In addition, the optic tracts and the tritocerebral giant neurons were stunted in parasitized individuals. Published evidence demonstrates changes in serotonin levels in hosts ranging from crustaceans to mammals infected by parasites as diverse as protozoans and helminths. The present study suggests that the degeneration of discrete sets of serotonergic neurons might underlie the serotonergic imbalance and thus contribute to host manipulation. PMID- 12769455 TI - Honeybee foraging in differentially structured landscapes. AB - Honeybees communicate the distance and location of resource patches by bee dances, but this spatial information has rarely been used to study their foraging ecology. We analysed, for the first time to the best of the authors' knowledge, foraging distances and dance activities of honeybees in relation to landscape structure, season and colony using a replicated experimental approach on a landscape scale. We compared three structurally simple landscapes characterized by a high proportion of arable land and large patches, with three complex landscapes with a high proportion of semi-natural perennial habitats and low mean patch size. Four observation hives were placed in the centre of the landscapes and switched at regular intervals between the six landscapes from the beginning of May to the end of July. A total of 1137 bee dances were observed and decoded. Overall mean foraging distance was 1526.1 +/- 37.2 m, the median 1181.5 m and range 62.1-10037.1 m. Mean foraging distances of all bees and foraging distances of nectar-collecting bees did not significantly differ between simple and complex landscapes, but varied between month and colonies. Foraging distances of pollen collecting bees were significantly larger in simple (1743 +/- 95.6 m) than in complex landscapes (1543.4 +/- 71 m) and highest in June when resources were scarce. Dancing activity, i.e. the number of observed bee dances per unit time, was significantly higher in complex than in simple landscapes, presumably because of larger spatial and temporal variability of resource patches in complex landscapes. The results facilitate an understanding of how human landscape modification may change the evolutionary significance of bee dances and ecological interactions, such as pollination and competition between honeybees and other bee species. PMID- 12769456 TI - Dancing for a decision: a matrix model for nest-site choice by honeybees. AB - A mathematical model is formulated for decision making by honeybees during nest site choice, using a population matrix model. This model explains how the observed dynamics of the nest-site scouts' dancing can reliably lead to a choice of the best nest site available. PMID- 12769457 TI - Global patterns in endemism explained by past climatic change. AB - I propose that global patterns in numbers of range-restricted endemic species are caused by variation in the amplitude of climatic change occurring on time-scales of 10-100 thousand years (Milankovitch oscillations). The smaller the climatic shifts, the more probable it is that palaeoendemics survive and that diverging gene pools persist without going extinct or merging, favouring the evolution of neoendemics. Using the change in mean annual temperature since the last glacial maximum, estimated from global circulation models, I show that the higher the temperature change in an area, the fewer endemic species of mammals, birds, reptiles, amphibians and vascular plants it harbours. This relationship was robust to variation in area (for areas greater than 10(4) km2), latitudinal position, extent of former glaciation and whether or not areas are oceanic islands. Past climatic change was a better predictor of endemism than annual temperature range in all phylads except amphibians, suggesting that Rapoport's rule (i.e. species range sizes increase with latitude) is best explained by the increase in the amplitude of climatic oscillations towards the poles. Globally, endemic-rich areas are predicted to warm less in response to greenhouse-gas emissions, but the predicted warming would cause many habitats to disappear regionally, leading to species extinctions. PMID- 12769458 TI - Genetic and plastic responses of a northern mammal to climate change. AB - Climate change is predicted to be most severe in northern regions and there has been much interest in to what extent organisms can cope with these changes through phenotypic plasticity or microevolutionary processes. A red squirrel population in the southwest Yukon, Canada, faced with increasing spring temperatures and food supply has advanced the timing of breeding by 18 days over the last 10 years (6 days per generation). Longitudinal analysis of females breeding in multiple years suggests that much of this change in parturition date can be explained by a plastic response to increased food abundance (3.7 days per generation). Significant changes in breeding values (0.8 days per generation), were in concordance with predictions from the breeder's equation (0.6 days per generation), and indicated that an evolutionary response to strong selection favouring earlier breeders also contributed to the observed advancement of this heritable trait. The timing of breeding in this population of squirrels, therefore, has advanced as a result of both phenotypic changes within generations, and genetic changes among generations in response to a rapidly changing environment. PMID- 12769460 TI - Rapid acquisition of an alarm response by a neotropical primate to a newly introduced avian predator. AB - Predation is an important selective pressure in natural ecosystems. Among non human primates, relatively little is known about how predators hunt primate prey and how primates acquire adaptive responses to counteract predation. In this study we took advantage of the recent reintroduction of radio-tagged harpy eagles (Harpia harpyja) to Barro Colorado Island (BCI), Panama to explore how mantled howler monkeys (Alouatta palliata), one of their primary prey, acquire anti predator defences. Based on the observation that harpies follow their prey prior to attack, and often call during this pursuit period, we broadcast harpy eagle calls to howlers on BCI as well as to a nearby control population with no harpy predation. Although harpies have been extinct from this area for 50-100 years, results indicate that BCI howlers rapidly acquired an adaptive anti-predator response to harpy calls, while showing no response to other avian vocalizations; howlers maintained this response several months after the removal of the eagles. These results not only show that non-human primates can rapidly acquire an alarm response to a newly introduced predator, but that they can detect and identify predators on the basis of acoustic cues alone. These findings have significant implications both for the role of learning mechanisms in the evolution of prey defence and for conservation strategies, suggesting that the use of 'probing' approaches, such as auditory playbacks, may highly enhance an a priori assessment of the impact of species reintroduction. PMID- 12769459 TI - Molecular phylogenetic evidence for ancient divergence of lizard taxa on either side of Wallace's Line. AB - Wallace's Line, separating the terrestrial faunas of South East Asia from the Australia-New Guinea region, is the most prominent and well-studied biogeographical division in the world. Phylogenetically distinct subgroups of major animal and plant groups have been documented on either side of Wallace's Line since it was first proposed in 1859. Despite its importance, the temporal history of fragmentation across this line is virtually unknown and the geological foundation has rarely been discussed. Using molecular phylogenetics and dating techniques, we show that the split between taxa in the South East Asian and the Australian-New Guinean geological regions occurred during the Late Jurassic to Early Cretaceous in two independent lizard clades. This estimate is compatible with the hypothesis of rifting Gondwanan continental fragments during the Mesozoic and strongly rejects the hypothetical origin of various members of the Australian-New Guinean herpetofauna as relatively recent invasions from South East Asia. Our finding suggests an ancient fragmentation of lizard taxa on either side of Wallace's Line and provides further evidence that the composition of modern global communities has been significantly affected by rifting and accretion of Gondwanan continental plates during the Middle to Late Mesozoic. PMID- 12769461 TI - Emergent Allee effects in top predators feeding on structured prey populations. AB - Top predators that forage in a purely exploitative manner on smaller stages of a size-structured prey population have been shown to exhibit an Allee effect. This Allee effect emerges from the changes that predators induce in the prey population size distribution and represents a feedback of predator density on its own performance, in which the feedback operates through and is modified by the life history of the prey. We demonstrate that these emergent Allee effects will occur only if the prey, in the absence of predators, is regulated by density dependence in development through one of its juvenile stages, as opposed to regulation through adult fecundity. In particular, for an emergent Allee effect to occur, over-compensation is required in the maturation rate out of the regulating juvenile stage, such that a decrease in juvenile density will increase the total maturation rate to larger/older stages. If this condition is satisfied, predators with negative size selection, which forage on small prey, exhibit an emergent Allee effect, as do predators with positive size selection, which forage on large adult prey. By contrast, predators that forage on juveniles in the regulating stage never exhibit emergent Allee effects. We conclude that the basic life-history characteristics of many species make them prone to exhibiting emergent Allee effects, resulting in an increased likelihood that communities possess alternative stable states or exhibit catastrophic shifts in structure and dynamics. PMID- 12769462 TI - Relatedness, body size and paternity in the alpine newt, Triturus alpestris. AB - Sexual selection has traditionally been investigated assuming that male quality is as skewed as patterns of male reproductive success can sometimes be. Recently, female choice has been investigated under the model of genetic compatibility, which assumes that each individual female has her own 'best' mate and there is no overall optimal choice for all females. We investigated female mate choice in the newt species Triturus alpestris, a member of a genus where female choice has been investigated only within the context of the optimal male (female choice for condition-dependent traits). We provided females with two males that differed in one condition-dependent trait (body size) and overall genetic composition. Both male body size and female body size did not influence paternity, but the degree of genetic relatedness between females and potential mates did. Two components of fitness (fecundity and hatching success) did not differ between singly and multiply sired clutches, indicating that females do not employ polyandry as a means of increasing offspring fitness through genetic bet-hedging. Instead, we hypothesize that females may mate initially for fertility assurance, but prefer less-related males as the most genetically compatible mates. PMID- 12769463 TI - Relative testis size and sperm morphometry across mammals: no evidence for an association between sperm competition and sperm length. AB - Understanding why there is extensive variation in sperm form and function across taxa has been a challenge because sperm are specialized cells operating at a microscopic level in a complex environment. This comparative study collates published data to determine whether the evolution of sperm morphometry (sperm total length and separate component dimensions) is associated with sperm competition (when different males' sperm mix and compete for a female's ova) across 83 mammalian species. We use relative testes mass as an indicator of the intensity of sperm competition across taxa: relative investment into testes is widely accepted to predict the level of sperm competition that a species or population endures. Although we found evidence for positive associations between relative testes mass (controlling for allometry) and sperm morphometry across 83 mammalian species, these relationships were phylogenetically dependent. When we appropriately controlled for phylogenetic association using multiple regression within a phylogenetic framework, there was no relationship between relative testes mass and sperm length across mammals. Furthermore, we found no evidence for associations between relative testes mass and sperm head, mid-piece or flagellar lengths, nor was there a relationship with mid-piece or mitochondrial volumes. Results, therefore, indicate that sperm competition does not select for longer or shorter sperm across mammals, and alternative forces selecting on sperm form and function are discussed. PMID- 12769464 TI - Overt and covert competition in a promiscuous mammal: the importance of weaponry and testes size to male reproductive success. AB - Male contests for access to receptive females are thought to have selected for the larger male body size and conspicuous weaponry frequently observed in mammalian species. However, when females copulate with multiple males within an oestrus, male reproductive success is a function of both pre- and postcopulatory strategies. The relative importance of these overt and covert forms of sexual competition has rarely been assessed in wild populations. The Soay sheep mating system is characterized by male contests for mating opportunities and high female promiscuity. We find that greater horn length, body size and good condition each independently influence a male's ability to monopolize receptive females. For males with large horns at least, this behavioural success translates into greater siring success. Consistent with sperm-competition theory, we also find that larger testes are independently associated with both higher copulation rates and increased siring success. This advantage of larger testes emerges, and strengthens, as the number of oestrous females increases, as dominant males can no longer control access to them all. Our results thus provide direct quantitative evidence that male reproductive success in wild populations of mammals is dependent upon the relative magnitude of both overt contest competition and covert sperm competition. PMID- 12769465 TI - Repeated inseminations required for natural fertility in a wild bird population. AB - In most bird species, pairs copulate many times before egg laying. The exact function of repeated inseminations (i.e. successful copulations) is unknown, but several suggestions have been made. We tested the hypothesis that repeated inseminations are required to ensure fertilization of eggs, by using an experimental method where free-ranging male collared flycatchers (Ficedula albicollis) were prevented from inseminating their mates. We show that egg fertility was lower when females had not copulated during the studied part of their fertile period. By counting sperm on the inner perivitelline layer of eggs, we estimated that a minimum of 86 sperm must reach the site of fertilization to ensure average fertility. Using the timing of inseminations and the numbers of sperm on successive eggs, we show that repeated copulations are necessary to achieve an average rate of fertilization of a single clutch. Our results thus provide evidence that repeated inseminations function to ensure fertilization success. We discuss possible constraints on sperm production and utilization that may have contributed to this pattern. PMID- 12769466 TI - A matter of taste: direct detection of female mating status in the bedbug. AB - Males of the bedbug, Cimex lectularius, traumatically inseminate females by inserting a needle-like intromittent organ (penis) through the female's abdominal wall after she has fed. We demonstrate that: (i) mating duration determines ejaculate size; (ii) a female's first copulation in a bout of copulations always lasts longer than subsequent copulations; (iii) the intromittent organ bears sensillae; (iv) males use their intromittent organ to 'taste' whether their current mate has recently copulated; and (v) the consequence of detecting female mating status is the reduction of copulation duration and ejaculate size. We discuss why male bedbugs might show this pattern of ejaculate-size adjustment. PMID- 12769467 TI - The evolution of mate choice and mating biases. AB - We review the current status of three well-established models (direct benefits, indirect benefits and sensory drive) and one newcomer (antagonistic chase-away) of the evolution of mate choice and the biases that are expressed during choice. We highlight the differences and commonalities in the underlying genetics and evolutionary dynamics of these models. We then argue that progress in understanding the evolution of mate choice is currently hampered by spurious distinctions among models and a misguided tendency to test the processes underlying each model as mutually exclusive alternatives. Finally, we suggest potentially fruitful directions for future theoretical and empirical research. PMID- 12769469 TI - IkappaB kinase and NF-kappaB signaling in response to pro-inflammatory cytokines. PMID- 12769471 TI - Adhesion molecule expression on cytokine-stimulated human endothelial cells. PMID- 12769470 TI - Screening for inhibitors of transcription factors using luciferase reporter gene expression in transfected cells. PMID- 12769472 TI - Phagocytosis by inflammatory phagocytes: experimental strategies for stimulation and quantification. PMID- 12769473 TI - Cytosolic Ca2+ measurement and imaging in inflammatory cells. PMID- 12769474 TI - Detection and visualization of oxidase activity in phagocytes. PMID- 12769475 TI - Measurement of complement activation. PMID- 12769476 TI - Measurement of matrix metalloproteinase activities in the medium of cultured synoviocytes using zymography. PMID- 12769477 TI - Measurement of aggrecanase-generated interglobular domain catabolites in the medium and extracts of cartilage explants using Western blot analysis. PMID- 12769478 TI - In vitro model of human articular cartilage degradation. PMID- 12769480 TI - Carrageenan-induced paw edema in the rat and mouse. PMID- 12769481 TI - Pleural models of inflammation: immune and nonimmune. PMID- 12769482 TI - Models of acute inflammation in the ear. PMID- 12769483 TI - Migration of specific leukocyte subsets in response to cytokine or chemokine application in vivo. PMID- 12769484 TI - Inflammatory joint disease: clinical, histological, and molecular parameters of acute and chronic inflammation and tissue destruction. PMID- 12769485 TI - The assessment of inflammation, cartilage matrix, and bone loss in experimental monoarticular arthritis of the rat. PMID- 12769486 TI - Collagen-induced arthritis. PMID- 12769487 TI - Air-pouch models of inflammation and modifications for the study of granuloma mediated cartilage degradation. PMID- 12769488 TI - Quantitative analysis of angiogenesis using the murine chronic granulomatous air pouch. PMID- 12769489 TI - Models of coronary artery occlusion and reperfusion for the discovery of novel antiischemic and antiinflammatory drugs for the heart. PMID- 12769490 TI - Assessment of anticolitic drugs in the trinitrobenzene sulfonic acid (TNBS) rat model of inflammatory bowel disease. PMID- 12769492 TI - In vivo models of inflammation: immune rejection and skin transplantation in vivo. PMID- 12769491 TI - An in vivo model of ischemia/reperfusion and inflammation of the kidneys of the rat. PMID- 12769493 TI - Wound healing: a model of dermal wound repair. PMID- 12769494 TI - An iconoclastic approach to pharmacodynamics in model systems: their relevance to humans. PMID- 12769495 TI - Quantifying inflammation in vivo using radiolabeled antibodies and leukocytes. PMID- 12769496 TI - Immunoperoxidase histochemistry for the detection of cellular adhesion molecule, cytokine, and chemokine expression in the arthritic synovium. PMID- 12769497 TI - Roles of nitric oxide and superoxide in inflammation. PMID- 12769498 TI - Analysis of nitrite and nitrate in the study of inflammation. PMID- 12769499 TI - In vivo assays for COX-2. PMID- 12769500 TI - Measurement of 8-epi-PGF2alpha as a marker of lipid peroxidation in vivo by immunoaffinity extraction and gas chromatography-mass spectrometry. PMID- 12769501 TI - Laboratory assessment of the acute phase response: using CRP as a model. PMID- 12769502 TI - Assays of matrix metalloproteinases (MMPs) and MMP inhibitors: bioassays and immunoassays applicable to cell culture medium, serum, and synovial fluid. PMID- 12769503 TI - Bendectin and birth defects: hopefully, the final chapter. PMID- 12769504 TI - Bendectin and birth defects. II: Ecological analyses. AB - BACKGROUND: Bendectin was the primary pharmaceutical treatment of nausea and vomiting of pregnancy (NVP) in the United States until the early 1980s. Its manufacture was then discontinued after public allegations that it was causing birth defects. Subsequently, meta-analyses of the many epidemiological cohort and case/control studies used to examine that hypothesis have demonstrated the absence of a detectable teratogenic effect. This study presents an ecological analysis of the same hypothesis that examines specific malformations. METHODS: Annual birth defect prevalence data for the 1970s to the 1990s have been obtained for specific birth defects from the Center for Disease Control's nationwide Birth Defect Monitoring Program. These data for the US have been compared graphically to the annual US Bendectin sales for the treatment of NVP. Data have also been obtained for annual US rates for hospitalization for NVP. The three data sets have been temporally compared in graphic analysis. RESULTS: The temporal trends in prevalence rates for specific birth defects examined from 1970 through 1992 did not show changes that reflected the cessation of Bendectin use over the 1980 84 period. Further, the NVP hospitalization rate doubled when Bendectin use ceased. CONCLUSIONS: The population results of the ecological analyses complement the person-specific results of the epidemiological analyses in finding no evidence of a teratogenic effect from the use of Bendectin. PMID- 12769505 TI - Teratogen-induced activation of the mitochondrial apoptotic pathway in the yolk sac of day 9 mouse embryos. AB - BACKGROUND: Using vital dyes, we have previously shown that while hyperthermia (HS), 4-hydroperoxycyclophosphamide (4CP), and staurosporine (ST) induce cell death within specific tissues (e.g., neuroepithelium) of day 9 mouse embryos, cells of the heart are resistant to the cell death-inducing potential of these teratogens. Subsequent work has shown that teratogen-induced cell death is associated with activation of the mitochondrial apoptotic pathway, i.e., release of cytochrome c from mitochondria, activation/cleavage of procaspase-9, -3, and 2, inactivation of poly(ADP-ribose) polymerase, and internucleosomal fragmentation of DNA, whereas resistance to teratogen-induced cell death in the heart is associated with a failure to activate this pathway. Teratogen-induced activation of the mitochondrial apoptotic pathway is initiated between 2.5 and 5 hr after teratogens are added to the culture medium. Because both the heart and the surrounding yolk sac are essential to successful development of mouse embryos during early postimplantation mouse development, we hypothesized that cells of the yolk sac are also resistant to teratogen-induced cell death. METHODS: To test our hypothesis, we cultured day 8.5 mouse conceptuses (embryo plus yolk sac) in whole embryo culture. On the morning of day 9, conceptuses were exposed to HS (43 degrees C for 15 min and then returned to 37 degrees C), 4CP (40 microM, 5-10 hr), or ST (0.5 microM 5-10 hr). At 5 and 10 hr after addition of teratogen, conceptuses were removed from culture and dissected into embryo and yolk sac. Activation of the mitochondrial apoptotic pathway was then assessed separately in embryos and yolk sacs using Western blot analysis to detect activation of procaspase-9, -3, and -2, enzyme assays to measure caspase-3-like activity, and immunohistochemistry to detect caspase-3 activation/cleavage in yolk sac cells. RESULTS: Although Western blot analysis revealed that procaspase-9, -3, and -2 were activated/cleaved in the embryo as early as the 5-hr time point, activation/cleavage of these caspases could not be detected in the yolk sac at either the 5- or 10-hr time point. Using an enzyme assay, we determined that caspase-3-like activity in the yolk sac was induced 1.7-fold by HS, 4.4-fold by 4CP, and 3.3-fold by ST. This compares to the embryo in which caspase-3-like activity was induced 45-fold by HS, 26-fold by 4CP, and 45-fold by ST. Using an antibody specific for the active p17 subunit of caspase-3 and immunohistochemistry, we were able to detect a small number of yolk sac cells showing caspase-3 activation. Thus, the low-level induction of caspase-3-like activity in the yolk sac is in part related to activation/cleavage of procaspase 3. CONCLUSIONS: Results presented indicate that cells of the extraembryonic yolk sac, like cells of the embryonic heart, are substantially more resistant to teratogen-induced activation of the mitochondrial apoptotic pathway and subsequent apoptosis compared to other embryonic tissues, particularly cells of the neuroepithelium. PMID- 12769506 TI - Exposure-disease continuum for 2-chloro-2'-deoxyadenosine, a prototype ocular teratogen. 3. Intervention with PK11195. AB - BACKGROUND: Treatment of pregnant mice with 2-chloro-2'-deoxyadenosine (2CdA) on Day 8 of gestation induces microphthalmia through a mechanism linked to the p53 tumor suppressor pathway. The present study defines the response of Day 8 mouse embryos through time with respect to pharmacologic intervention with PK11195, a ligand of the mitochondrial peripheral benzodiazepine receptor (Bzrp). METHODS: Pregnant CD-1 mice dosed with 2CdA with or without PK11195 on gestation Day 8 provided fetuses for teratologic evaluation on Day 14 and Day 17; HPLC measured pyridine nucleotides (NADH/NAD+) at 1.5 hr, RT-PCR measured mitochondrial 16S rRNA abundance at 3.0 hr, and p53 protein induction was assessed with immunostaining at 4.5 hr postexposure. RESULTS: The mean incidences of malformed fetuses were significantly higher in the 7.5 mg/kg 2CdA treatment group (50.2% malformed) vs. the 2CdA + 4.0 mg/kg PK11195 co-treatment group (4.4% malformed). Malformed fetuses displayed a range of ocular defects that included microphthalmia and keratolenticular dysgenesis (Peters anomaly). No malformations were observed in the control or PK11195 alone groups. PK11195 also protected litters from increased resorption rates and fetal weight reduction. It did not rescue early effects on NADH balance (1.5 hr) or 16S rRNA expression (3.0 hr); however, the p53 response (4.5 hr) was downgraded in 2CdA + PK11195 embryos vs. 2CdA alone. By delaying the administration of PK11195 in 1.5 hr intervals it was determined that the window for protection closed between 4.5 to 6.0 hr after 2CdA. CONCLUSIONS: The capacity of PK11195 to suppress the pathogenesis of microphthalmia implies a critical role for mitochondrial peripheral benzodiazepine receptors in the p53-dependent mode of action of 2CdA on ocular development. PMID- 12769507 TI - Pathogenic pathways in fluconazole-induced branchial arch malformations. AB - BACKGROUND: A widely-used antimycotic agent, bis-triazole fluconazole (FLUCO), is able to produce abnormalities to the branchial apparatus (hypoplasia, agenesis, and fusion) in postimplantation rodent embryos cultured in vitro. The branchial apparatus is a complex and transient structure in vertebrate embryos and is essential for the development of the face skeleton. Branchial arch mesenchyme is formed by two different cellular populations: paraxial mesenchyme and ectomesenchyme, which originate from rhombencephalic neural crest cell (NCC) migration. We investigated the possible pathogenic pathways involved in FLUCO related branchial arch abnormalities. Perturbations in physiological apoptosis, cell proliferation, NCC migration and branchial mesenchyme induction have been considered. METHODS: Rat embryos (9.5-day postcoitum; 1-3 somites) were exposed in vitro to 0 or 500 microM FLUCO. After 24, 36, or 48 hr of culture, embryos were examined for apoptosis (acridine orange method) and cell proliferation (BrdU incorporation and detection method). Rhombencephalic NCC migration was analyzed using immunostaining of NCC (using anti-CRABP antibodies) and the extracellular matrix (using anti-fibronectin antibodies). The differentiative capability of the branchial mesenchymes was investigated using anti-endothelin and anti-endothelin receptor antibodies. RESULTS: During the whole culture period, no alterations in physiological apoptosis, cell proliferation, and mesenchymal cell induction were observed in FLUCO-exposed embryos in comparison to controls. On the contrary, severe alterations in NCC migration pathways were observed in FLUCO-exposed embryos. CONCLUSIONS: The findings suggest that FLUCO produces teratogenic effects by interfering with the cellular and molecular mechanisms that control NCC migration. PMID- 12769508 TI - Birth prevalence and pattern of osteochondrodysplasias in an inbred high risk population. AB - BACKGROUND: Define the pattern and birth prevalence of the different types of osteochondrodysplasias in newborn infants in the United Arab Emirates (UAE) population, which is highly inbred and where termination of pregnancy is not accepted. METHODS: All infants with a birth weight of 500 gm and above in the three hospitals in Al Ain Medical District of the UAE were studied prospectively over a period of 5 years. For each live birth or stillbirth with suspected skeletal dysplasia, a detailed clinical and radiological examination was carried out. Pregnancy history and information regarding parental age, ethnic origin, family history, and level of consanguinity were obtained and a pedigree was constructed. RESULTS: Among the 38,048 births during the study period, 36 (9.46/10,000 births) had some type of skeletal dysplasia. Eighteen cases were attributed to autosomal recessive genes (4.7/10,000 births), 10 were due to apparent new dominant mutations (2.62/10,000), five were autosomal dominant type (1.3/10,000) and one was X-linked dominant type (0.26/10,000). In three cases, inheritance was unknown. The most common recessive type of skeletal dysplasia in our series was fibrochondrogenesis (1.05/10,000), followed by chondrodysplasia punctata (0.78/10,000). The birth prevalence rate of skeletal dysplasia doubled in the last 2 years of the 5-year observation period (6.74/10,000 in 1996 vs. 12.86/10,000 in 1999, and 13.45/10,000 in 2000). This increase involved cases caused by new dominant mutations, and occurred mainly in the first half of 1999. CONCLUSION: This prospective study has identified a high birth prevalence of skeletal dysplasia, and risk factors are postulated. These findings represent an accurate birthprevalence figure and a useful baseline for this group of birth defects in the UAE. PMID- 12769509 TI - Are there teratogenic risks associated with antidotes used in the acute management of poisoned pregnant women? AB - OBJECTIVE: We reviewed evidence suggesting teratogenic risk associated with the use of antidotes in the acute management of poisoned pregnant women. METHODS: Medline, Toxline, and DART/ETIC searches; references of retrieved articles, pertinent databases and textbooks were also searched. RESULTS: There are case reports or case series of women who received antidotes for poisoning during (*) or after (+) the period of organogenesis who showed no fetal adverse effects. Some antidotes, however, have no teratogenic risk: atropine (cohort/surveillance studies)+, calcium (oral supplement: cohort study)+ and pyridoxine (Bendectin studies). Also, ethanol+, methylene blue (intra-amniotic injection but not oral) and penicillamine* can be considered teratogens but their risks in the treatment of poisonings are unknown. There is no epidemiologic study evaluating the risk of the following antidotes during pregnancy: N-acetylcysteine(*+), BAL (dimercaprol)+, black widow spider antivenin+, calcium EDTA+, crotalidae antivenin, crotalidae polyvalent immune FAB, cyanide antidote kit (amyl and sodium nitrate, sodium thiosulfate), deferoxamine(*+), digoxin immune FAB+, DMSA+, flumazenil+, fomepizole, methylene blue (IV), naloxone, physostigmine, pralidoxime+, protamine+; and parenteral pharmacologic doses of calcium+, folinic acid*, glucagon+, hydroxycobalamin, phytonadione (vitamin K), and pyridoxine. CONCLUSIONS: Despite the limited evidence supporting the risk of antidote use during pregnancy, antidotes should be used when there is a clear maternal indication to decrease the morbidity or mortality associated with poisoning. The only exception may be penicillamine, which is a teratogen. Better antidotes exist for most poisonings that penicillamine could potentially treat. At this time, there is no known fetal indication for all antidotes. Reporting the use of an antidote during pregnancy should be encouraged, especially if used during the critical period of organogenesis. PMID- 12769510 TI - Narrating troubling experiences. AB - This article presents a process-oriented perspective that relates to the broad question of how self-related experience comes to be endowed with meaning. The approach highlights the implications of 'living by' particular culturally based understandings in specific contexts and centers on how jointly cultural, social, and cognitive processes offer potentialities for orienting the experiential self without determining self-related experiences. This process-oriented perspective revolves around the interplay between the range of historically contingent cultural resources available for endowing experience with meaning and the socially and structurally grounded processes through which individuals learn about, orient towards and traffic in interpretive plausibilities--a socially situated experientially based process. This perspective is informed by, and provides an entree for exploring, variability within a cultural setting. The narrative accounts examined are from individuals who grew up speaking either Ojibwa or Cree (both Algonkian languages) in First Nations communities in Manitoba, Canada. PMID- 12769511 TI - Acculturation and changing concepts of mental disorder: Brazilians in the USA. AB - Forty-three Brazilian citizens living in the USA judged whether a sample of conditions were mental disorders and rated them on proposed features of the concept of mental disorder. Judgments and ratings were correlated with measures of American acculturation and identification with Brazilian culture, and with years of American residence. Consistent with prediction, greater acculturation was associated with a concept of disturbio mental that was broader in reach and more intrapsychic in focus. However, greater acculturation was also associated with a stronger tendency to understand disorder as a violation of social expectations and to pathologize behavior in excess or 'acting out.' American acculturation yielded no convergence of disturbio mental with the concept of disorder embodied in DSM-IV. PMID- 12769512 TI - Are children with imaginary playmates and children said to remember previous lives cross-culturally comparable categories? AB - Some 15-66% of children in western countries talk about imaginary playmates; in India only 0.2% of children are said to remember a previous life. Both phenomena occur from the age of 30 months to 60-90 months. This article explores whether the two phenomena are cross-culturally comparable categories. The article describes a study of the psychological characteristics of a sample of 15 children said to remember a previous life in India, compared with a matched sample; and compares it with a sample of 15 children with imaginary playmates and a matched sample from Charlottesville, Virginia. No significant differences between the target group and the comparison group were found in either culture, suggesting that both phenomena are normal. One case of an American child with an imaginary playmate and one case of a child in India who is said to remember a previous life are described, using a video-recording of the child's dialogue. PMID- 12769513 TI - Pre-migration trauma and HIV-risk behavior. AB - This study examined the relationship between pre-migration trauma and HIV-risk behavior in refugees from sub-Saharan Africa. The sample comprised 122 persons who had emigrated from sub-Saharan Africa and were currently residing in Sweden. Qualitative methods including individual interviews, focus groups, and interviews with key informants addressed questions regarding trauma experience and HIV-risk behavior. A history of pre-migration trauma was found to be associated with HIV risk behavior. According to the participants, symptoms associated with post traumatic stress disorder, depression, adjustment disorder, and substance use mediated the relationship between pre-migration trauma and sexual risk behavior. In contrast, a minority of the participants who reported pre-migration trauma but not psychological sequelae, or experienced post-traumatic growth, reported safer sexual practices. It appears that for some individuals, pre-migration trauma resulted in psychiatric sequelae, which may increase an individual's risk to be infected with HIV. Interventions targeted at individuals at increased risk (i.e. pre-migration trauma with unresolved psychiatric symptomatology) may facilitate the prevention of HIV and other sexually transmitted diseases in this population. Integration of multiple psychosocial and health issues is recommended for comprehensive treatment and prevention programs. PMID- 12769514 TI - On crafting a cultural mind: a comparative assessment of some recent theories of 'internalization' in psychological anthropology. AB - This article reviews a number of recent publications in psychological anthropology that draw in varying degrees from psychoanalytic premises in order to theoretically address problems concerning the internalization of cultural meaning. The article begins with a discussion and critical comparison of Spiro's and Obeyesekere's perspectives on internalization that are in line with a number of classical formulations in anthropological and psychoanalytic theory, before turning to explore what appears to be an emerging new wave of perspectives in contemporary psychological anthropology that set out to discuss problems of internalization in the context of a complex modeling of psychological, social, and cultural processes. The article concludes with a brief discussion of where researchers may need to turn to further our understanding of 'internalization' in relation to those intrapsychic, interpsychic, and extrapsychic processes underpinning the crafting of cultural minds. PMID- 12769515 TI - Accurate determination of 2,4,6-trichloroanisole in wines at low parts per trillion by solid-phase microextraction followed by GC-ECD. AB - A headspace solid-phase microextraction (HS-SPME) procedure at 30 degrees C with a 100 microm PDMS fiber of a saturated NaCl solution stirred at 1100 rpm combined to GC-ECD for the 2,4,6-trichloroanisol (TCA) determination in wines has been developed. Due to the matrix complexity and ethanol absorption into the fiber, the internal standard selection was crucial to obtain unbiased results. Thus, matrix effects were observed when analyzing different types of Spanish wines (white, early, and vintage red wines) spiked with TCA at low concentration levels (i.e., <40 ng L(-)(1)). In contrast, the use of 2,4,6-tribromoanisole (TBA) as internal standard overcame these matrix effects, whereas the use of 2,4,6 trichlorophenyl ethyl ether led to inconsistent results. The developed HS-SPME-GC ECD methodology reaches a limit of quantitation for TCA in wine within 2.9-18 ng L(-)(1), with a relative standard deviation of 2.5-13.4%, depending on the TCA concentration level and wine characteristics. This analytical method is comparable to the existing methodologies based on HS-SPME followed by GC-MS in terms of accuracy, precision, length of determination, and length of quantification; however, analysis cost is reduced. PMID- 12769516 TI - Virgin olive oil quality classification combining neural network and MOS sensors. AB - A model based on neural networks has been designed to detect lampante virgin olive oils, a category of olive oil that cannot be consumed without a previous refining process according to the current regulation of the European Communities. The response of 7 metal oxide sensors analyzing 114 olive oil samples has been used in the design, training, and internal validation of the neural network with only 4.5% error in validation. The designed mathematical model, the equations of which are fully described, has been validated also with an external set of 13 samples of diverse varieties and geographical origins with 100% correct classification. PMID- 12769517 TI - Simultaneous identification of soyasaponins and isoflavones and quantification of soyasaponin Bb in soy products, using liquid chromatography/electrospray ionization-mass spectrometry. AB - A method has been developed for simultaneous identification of soyasaponins and soy isoflavones in soy products, based on liquid chromatography/electrospray ionization-mass spectrometry (LC/ESI-MS). Soy-based nutraceutical products were analyzed by LC/ESI-MS with detection of protonated and sodiated molecular ions, as well as characteristic fragment ions for these compounds. Soy isoflavones were characterized by a strong protonated molecular ion in addition to corresponding [aglycone + H](+) ions. Monitoring the soyasaponin-specific protonated aglycone and dehydrated aglycone ions throughout the chromatogram provided a unique fingerprint for soyasaponin content in the samples. This mass spectrometric fingerprint also allowed immediate classification of the soyasaponin analytes as group A or B soyasaponins, based on the unique masses of aglycone ions observed for each class. Quantification of soyasaponin B(b) in soy-derived materials, based on the use of a purified soyasaponin B(b) standard and a glycyrrhizin internal standard, has been accomplished. PMID- 12769518 TI - Soil transformation of prosulfuron. AB - The transformation of prosulfuron [1-(4-methoxy-6-methyltriazine-2-yl)-3-[2 (3,3,3-trifluropropyl)phenylsulfonyl]urea] in three soils at different pH values (sterilized and unsterilized) was studied, and it was shown that the rate of transformation was high in acidic soil. From the results obtained in sterile soils, it is shown that the mechanism of dissipation was mainly chemical in acidic soils. A new metabolite, 2-(3,3,3-trifluoropropyl)phenylsulfonic acid, was identified. PMID- 12769519 TI - Flavonoids detection by HPLC-DAD-MS-MS in lemon juices from Sicilian cultivars. AB - High-performance liquid chromatography-diode array detector (HPLC-DAD) coupled with electronspray mass spectrometry (ESI-MS) was used to detect the flavonoid profile in lemon juices obtained from the main Sicilian cultivars (Femminello comune, Monachello, and Interdonato). Significant amounts of an unusual constituent were found in the lemon juice of the above-mentioned cultivars together with eriocitrin, hesperidin, and diosmin. Following purification by preparative HPLC, the structure was assigned as 6,8-di-C-glucopyranosyldiosmetin by means of DAD-UV, ESI-MS-MS, and (1)H NMR analysis. Three other minor components were also detected. One of these presented a flavone nature, and spectral data and literature references both suggested a 6,8-di-C glucopyranosylapigenin structure. The different contents of eriocitrin, hesperidin, diosmin, and above minor components in the cultivars allow juices to be readily differentiated. PMID- 12769520 TI - Elucidation of lignin structure through degradative methods: comparison of modified DFRC and thioacidolysis. AB - Milled wood and milled wood lignin (MWL) samples were subjected to DFRC and thioacidolysis. Despite the fact that both methods selectively cleave aryl ether bonds, substantial differences in results were obtained. Lignin thioacidolysis gave total molar yields of degradation monomer products in the range of 3.5-7 mol % higher than DFRC. GPC analysis showed that the thioacidolysis-treated lignin was degraded to a lower average molecular weight than that treated by DFRC. Contrary to results reported for lignin model compounds, these results indicate that the DFRC method does not completely or efficiently degrade the lignin polymer. In fact, the DFRC-degraded lignin retained much of the characteristics of the original MWL. Elemental analysis revealed the presence of bromine in the DFRC-treated lignin, and two-dimensional (1)H-(13)C HMQC NMR spectroscopy showed the presence of beta-O-4 linkages in the DFRC-treated lignin. No beta-O-4 interunit linkages were detected in the thioacidolysis-treated lignin. These results are consistent with the lower monomer yields and the higher average molecular weight of the DFRC-treated lignin and indicate inefficiency in the chemistry of the method, probably due to steric constraints of the polymeric nature of lignin. PMID- 12769521 TI - Identification of triterpene hydroxycinnamates with in vitro antitumor activity from whole cranberry fruit (Vaccinium macrocarpon). AB - Bioactivity-guided fractionation of cranberry fruit was used to determine the identity of triterpenoid esters from Vaccinium macrocarpon, which inhibit tumor cell growth and may play a role in cancer prevention. In our previous study, a fraction from whole fruit exhibited tumor cell growth inhibition in vitro. The major components of this fraction were isolated by chromatographic separation of ethyl acetate extracts, purified by semipreparative HPLC, and identified by NMR as cis- (1) and trans- (2) isomers of 3-O-p-hydroxycinnamoyl ursolic acid. These triterpenoid esters have not been previously reported in Vaccinium fruit. Bioassay of the purified triterpene cinnamates in tumor cell lines in vitro showed slightly greater activity of compound 1 in most cell lines, with GI(50) values of approximately 20 microM in MCF-7 breast, ME180 cervical and PC3 prostate tumor cell lines. Quercetin was slightly less active than 1, while cyanidin-3-galactoside exhibited much lower cytotoxicity, with GI(50) greater than 250 microM in all cell lines. Phenylboronic acid (3) was also isolated from the fruit but showed insignificant antitumor activity. PMID- 12769522 TI - Profiling glucosinolates and phenolics in vegetative and reproductive tissues of the multi-purpose trees Moringa oleifera L. (horseradish tree) and Moringa stenopetala L. AB - Moringa species are important multi-purpose tropical crops, as human foods and for medicine and oil production. There has been no previous comprehensive analysis of the secondary metabolites in Moringa species. Tissues of M. oleifera from a wide variety of sources and M. stenopetala from a single source were analyzed for glucosinolates and phenolics (flavonoids, anthocyanins, proanthocyanidins, and cinnamates). M. oleifera and M. stenopetala seeds only contained 4-(alpha-l-rhamnopyranosyloxy)-benzylglucosinolate at high concentrations. Roots of M. oleifera and M. stenopetala had high concentrations of both 4-(alpha-l-rhamnopyranosyloxy)-benzylglucosinolate and benzyl glucosinolate. Leaves from both species contained 4-(alpha-l-rhamnopyranosyloxy) benzylglucosinolate and three monoacetyl isomers of this glucosinolate. Only 4 (alpha-l-rhamnopyranosyloxy)-benzylglucosinolate was detected in M. oleifera bark tissue. M. oleifera leaves contained quercetin-3-O-glucoside and quercetin-3-O (6' '-malonyl-glucoside), and lower amounts of kaempferol-3-O-glucoside and kaempferol-3-O-(6' '-malonyl-glucoside). M. oleifera leaves also contained 3 caffeoylquinic acid and 5-caffeoylquinic acid. Leaves of M. stenopetala contained quercetin 3-O-rhamnoglucoside (rutin) and 5-caffeoylquinic acid. Neither proanthocyanidins nor anthocyanins were detected in any of the tissues of either species. PMID- 12769523 TI - Analysis of isothiocyanate mercapturic acids in urine: a biomarker for cruciferous vegetable intake. AB - Cruciferous vegetables contain glucosinolates, which are degraded to isothiocyanates. These are easily absorbed, conjugated to glutathione, and excreted into the urine as their corresponding mercapturic acids. We have developed and validated a solid phase extraction-high-performance liquid chromatography-electrospray ionization mass spectrometry/mass spectrometry method for the specific analysis of individual isothiocyanate mercapturic acids in urine. The range of reliable analysis was 1.0-310 microM in urine. Urine samples fortified with three different levels of isothiocyanate mercapturic acids were measured on six different days by three independent technicians. The relative standard deviation (RSD) of repeatability was 12, 6, and 3%; the RSD of reproducibility was 19, 14, and 8%, and spike recoveries were 103, 104, and 103%, respectively, for 1.04, 10.5, and 313 microM levels. In 24 h urine collected from two volunteers after they consumed broccoli and cauliflower, clearly sulforaphane mercapturic acid (133 micromol) and allyl isothiocyanate mercapturic acid (4.7 micromol) were found. This procedure demonstrates a reliable and efficient method to study the intake and mode of action of isothiocyanates in animal studies and clinical trials. PMID- 12769524 TI - Stimulatory effect of cyanidin 3-glycosides on the regeneration of rhodopsin. AB - Anthocyanins have been suggested to improve visual functions. This study examined the effect of four anthocyanins in black currant fruits on the regeneration of rhodopsin using frog rod outer segment (ROS) membranes. Cyanidin 3-glycosides, glucoside and rutinoside, stimulated the regeneration, but the corresponding delphinidins showed no significant effect. The formation of a regeneration intermediate was suggested to be accelerated by cyanidin 3-rutinoside. Their effects on the cGMP-phosphodiesterase activity in the ROS membranes were also investigated but found to be negligible. It was concluded that the major effect of anthocyanins in rod photoreceptors is on the regeneration of rhodopsin. PMID- 12769525 TI - Antimutagenicity of supercritical CO2 extracts of Terminalia catappa leaves and cytotoxicity of the extracts to human hepatoma cells. AB - Natural antimutagens may prevent cancer and are therefore of great interest to oncologists and the public at large. Phytochemicals are potent antimutagen candidates. When the Ames test was applied to examine the antimutagenic potency of supercritical carbon dioxide (SC-CO(2)) extracts of Terminalia catappa leaves at a dose of 0.5 mg/plate, toxicity and mutagenicity were not detected. The antimutagenic activity of SC-CO(2) extracts increased with decreases of temperature (60, 50, and 40 degrees C) and pressure (4000, 3000, and 2000 psi) used for extraction. The most potent antimutagenicity was observed in extracts obtained at 40 degrees C and 2000 psi. At a dose of 0.5 mg of extract/plate, approximately 80% of the mutagenicity of benzo[a]pyrene (B[a]P, with S-9) and 46% of the mutagenicity of N-methyl-N '-nitroguanidine (MNNG, without S-9) were inhibited. Media supplemented with SC-CO(2) extracts at a range of 0-500 microg/mL were used to cultivate human hepatoma (Huh 7) and normal liver (Chang liver) cells. The viability of the cells was assayed by measuring cellular acid phosphatase activity. A dose-dependent growth inhibition of both types of cells was observed. The SC-CO(2) extracts were more cytotoxic to Huh 7 cells than to Chang liver cells. The observation that SC-CO(2) extracts of T. catappa leaves did not induce mutagenicity at the doses tested while exhibiting potent antimutagenicity and were more cytotoxic to human hepatoma cells than to normal liver cells is of merit and warrants further investigation. PMID- 12769526 TI - Enhanced resistance to Helicoverpa zea in tobacco expressing an activated form of maize ribosome- inactivating protein. AB - Progeny of two transgenic tobacco (Nicotiana tabacum L.) lines that expressed an activated form of maize (Zea mays L.) ribosome-inactivating protein (RIP) had varying resistance to the insect species tested. A subset of R(2) plants from the two lines appeared to be more resistant to larvae of the cigarette beetle, Lasioderma serricorne (F.), and the tobacco hornworm, Manduca sexta (L.) than the wild type plants. Progeny (R(3)) of the more resistant R(2) plants were tested more extensively for insect resistance. Resistance to the corn earworm, Helicoverpa zea (Boddie), was most consistent, with significantly decreased feeding often accompanied by increased mortality and reduced weights of survivors fed on leaf disks of the two transgenic lines compared to the wild type. The amount of damage by H. zea was significantly inversely correlated with levels of RIP. Resistance of RIP-producing plants to H. zea was greater than expected on the basis of prior in vitro results using diet-incorporated maize RIP. The R(3) transgenic plant leaf disks were also often more resistant to feeding by larvae of L. serricorne compared to wild type plants. Although reduced feeding by M. sexta was noted when they were fed leaf disks from transgenic compared to wild type plants the first day of exposure, differences were not significant. This information provides further support for maize RIP having a role in resistance to maize-feeding insects. PMID- 12769527 TI - Changes in the chemical composition of basil caused by different drying procedures. AB - Basil (Ocimum basilicum L.) leaves were dried using a microwave oven at atmospheric pressure or two traditional methods: air-drying at 50 degrees C and freeze-drying. The microwave-drying was carried out at different powers and times on raw basil leaves, while for air and freeze-drying techniques, both raw and blanched leaves were used. The raw and dried basil was analyzed for selected aroma compounds by gas chromatography/mass spectrometry-selected-ion-monitoring, the chlorophyll a and b by HPLC and the color by a reflected-light colorimeter. For dried samples microwaved for 1 min at 270, 2 min at 440, 1 min at 650, and 1 min at 1100 W, the percentage retentions of the characteristic volatile compounds (eucalyptol, linalool, eugenol, and methyl eugenol) were higher than in the samples dried by traditional methods, with the exception of freeze-dried unblenched basil. Microwave drying allowed a larger retention of chlorophyll pigments than air-drying and freeze-drying (with or without blanching) and preserved the color of the raw basil. Microwave drying requires a much shorter treatment and implied the simultaneous blanching of the material. PMID- 12769528 TI - The effects of frozen storage conditions on lycopene stability in watermelon tissue. AB - The purpose of this investigation was to evaluate the rate of deterioration of lycopene in watermelon tissue during frozen storage, because little is known about the stability of watermelon tissue lycopene under cold storage conditions. Heart tissue from each of nine individual watermelons was stored at -20 or -80 degrees C as either small chunks or puree and periodically sampled over a year's time. Initial freeze-thaw experiments indicated that a small percentage of lycopene, approximately 4-6%, degraded during an initial freeze-thaw. Analyses of the samples showed a loss of approximately 30-40% lycopene over a year's storage at -20 degrees C and a loss of approximately 5-10% over the same period at -80 degrees C. Lycopene was slightly more stable in pureed compared with diced watermelon tissue at -20 degrees C, but not at -80 degrees C. The kinetic data were best fitted by application of two simultaneous, first-order decay processes. HPLC analysis of the samples after a year's storage suggested that beta-carotene was more stable during storage at -20 degrees C than was lycopene. PMID- 12769529 TI - Comparison of isothiocyanate yield from wasabi rhizome tissues grown in soil or water. AB - The isothiocyanate (ITC) yield of wasabi, the Japanese horseradish (Wasabi japonica), was measured on its release from glucosinolates in the rhizomes of plants grown in two traditional ways. Mature plants of 18 months old were harvested from two different commercial farms located in the South Island of New Zealand. At one farm, the plants were grown in raised soil beds, while the plants at the other farm were grown in gravel irrigated by river water. Following harvest, the rhizomes from each growth medium were divided into five size groups based on the weight and length of the rhizomes. The different sized rhizomes were also subdivided into proximal, medial, and distal portions of the rhizomes and each portion was further subdivided into epidermis plus cortex, and vascular plus pith. The individual and total ITC contents of each portion (proximal, medial, and distal) of the rhizomes were measured using dichloromethane extraction followed by the GC-FPD. The total ITC content of the rhizomes grown in soil increased (13 times) linearly from 6 to 114 g of rhizome weight, while the mean ITC content of the water-grown wasabi increased (10 times) nonlinearly for similar sized rhizomes. Water-grown rhizomes in the weight range from 18 to 45 g gave significantly (P = 0.030) higher total ITC (1-2 times) than similarly sized soil-grown rhizomes. Analysis of the tissues showed that the total and the individual ITCs were found in significantly higher levels (73 and 64%, respectively) in the skin and cortex tissue compared to the vascular and pith tissues. Analysis of the ITC content of the different locations of the wasabi rhizome showed that the distal portion of the rhizome contained significantly higher levels of both total and individual ITCs compared to the medial and proximal portions of the rhizome. PMID- 12769530 TI - Characterization of a colorless anthocyanin-flavan-3-ol dimer containing both carbon-carbon and ether interflavanoid linkages by NMR and mass spectrometry. AB - Direct addition of anthocyanins and flavan-3-ols was investigated in a model system by incubating malvidin 3-glucoside and (-)-epicatechin in ethanol. Analysis of reaction products by high-performance liquid chromatography coupled to electrospray ionization mass spectrometry (HPLC/ESI-MS) before and after thiolysis showed the formation of colorless dimers detected at m/z 781 in the negative ion mode, with retention times and spectroscopic characteristics identical to those of compounds detected in wine, which contain one malvidin 3 glucoside unit and one flavanol unit. On the basis of their resistance to thiolysis, these compounds were postulated to be bicyclic dimers linked with both carbon-carbon and ether bonds as observed in the case of A type proanthocyanidins. The major dimer analyzed by NMR experiments was identified as malvidin 3-glucoside(C2-O-C7,C4-C8)epicatechin, confirming this hypothesis. A similar assay was performed with (+)-catechin instead of (-)-epicatechin, and the formation of bicyclic dimers was also observed. PMID- 12769531 TI - Metsulfuron methyl sorption-desorption in field-moist soils. AB - Pesticide sorption coefficients (K(d)) are generally obtained using batch slurry methods. As a consequence, the results may not adequately reflect sorption processes in field-moist or unsaturated soil. The objective of this study was to determine sorption of metsulfuron methyl, a weak acid, in field-moist soils. Experiments were performed using low density (i.e., 0.3 g mL(-)(1)) supercritical fluid carbon dioxide (SF-CO(2)) to convert anionic metsulfuron methyl to the molecular species and remove it from the soil water phase only, thus allowing calculation of sorption coefficients (K(d)) at low water contents. K(d) values for sorption of the metsulfuron methyl molecular species on sandy loam, silt loam, and clay loam soil at 11% water content were 120, 180, and 320 mL g(-)(1), respectively. Using neutral species K(d) values, the pK(a) of metsulfuron methyl, and the pH of the soil, we could successfully predict the K(d) values obtained using the batch slurry technique, which typically has a predominance of anionic species in solution during the sorption characterization. This application of supercritical fluid extraction to determine sorption coefficients, combined with sulfonylureas' pK(a) values and the soil pH, will provide an easy method to predict sorption in soil at different pH levels. PMID- 12769532 TI - Sorption-desorption of two "aged" sulfonylaminocarbonyltriazolinone herbicide metabolites in soil. AB - Aging (herbicide-soil contact time) has been shown to significantly affect the sorption-desorption characteristics of many herbicides, which in turn can affect the availability of the herbicide for transport, plant uptake, and microbial degradation. In contrast, very little work in this area has been done on herbicide metabolites in soil. The objective of this study was to characterize the sorption-desorption of sulfonylaminocarbonyltriazolinone herbicide metabolites incubated in soils at different soil moisture potentials. A benzenesulfonamide metabolite and a triazolinone metabolite from sulfonylaminocarbonyltriazolinone herbicides were incubated in clay loam and loamy sand soils for up to 12 weeks at -33 kPa and at water contents equivalent to 50 and 75% of that at -33 kPa. Chemicals were extracted sequentially with 0.01 N CaCl(2) and aqueous acetonitrile (solution and sorbed phase concentrations, respectively), and apparent sorption coefficients (K(d,app)) were calculated. Sufficient metabolite remained during the incubation (>55% of applied) to allow determination of the coefficients. The initial aging period (2 weeks after application) significantly increased sorption as indicated by increased K(d,app) values for the chemical remaining, after which they remained relatively constant. After 12 weeks of incubation at -33 kPa, K(d,app) values for benzenesulfonamide and triazolinone increased by a factor of 3.5 in the clay loam soil and by a factor of 5.9 in the loamy sand as compared to freshly treated soils. There was no effect of moisture potential on aged apparent K(d,app) values. These data show the importance of characterization of sorption-desorption in aged herbicide residues, including metabolites, in soil, particularly in the case of prediction of herbicide residue transport in soil. In this case, potential transport of sulfonylaminocarbonyltriazolinone herbicide metabolites would be overpredicted if freshly treated soil K(d) values were used to predict transport. PMID- 12769533 TI - Fate of selenate metabolized by Enterobacter taylorae isolated from rice straw. AB - Rice straw has been successfully tested as an effective organic source and a carrier of selenate [Se(VI)]-reducing bacteria to remove Se(VI) from agricultural drainage water. In this study, an Se(VI)-reducing bacterium identified as Enterobacter taylorae was isolated from rice straw and used to remove Se(VI) from a 0.5% tryptic soy broth (TSB) and high-salt (15.5 dS m(-)(1)) synthetic agricultural drainage water containing Se(VI) in a range of 500-5000 microg/L. Results showed that E. taylorae reduced 81-94% of the added Se(VI) to elemental Se [Se(0)] in the 0.5% TSB solution during a 5-day experiment. In the high-salt drainage water, Se(VI) reduction was rapid during a 9-day experiment. On the final day of the experiment, Se(0) [75%] and Se(-II) [19%] were the major forms of Se in the drainage water with small amounts of Se(VI), Se(IV), and volatile Se released. The pathway of Se(VI) reduction in the drainage water followed the order Se(VI) --> selenite [Se(IV)] --> Se(0) --> selenide [Se(-II)]. This study suggests that E. taylorae may be used to remediate high-salt Se(VI)-contaminated agricultural drainage water. PMID- 12769534 TI - Composition of the essential oil of Mentha microphylla from the Gennargentu Mountains (Sardinia, Italy). AB - The essential oil obtained by hydrodistillation of the fresh aerial parts of Mentha microphylla C. Kock (Lamiaceae) collected on the Gennargentu Mountains (Sardinia, Italy) has been investigated by gas chromatography (GC) and GC/mass spectrometry (MS). The main constituents that resulted were pulegone (34.1%), piperitenone oxide (32.9%), and piperitenone (11.3%). The presence of small amounts of compounds such as ethyl hexanoate, 1-octen-3-ol, nonanal, and ethyl 2 methylbutanoate could justify the particular odorous profile of the plant, resembling the aroma of milk and other dairy products such as mozzarella. PMID- 12769535 TI - Combinatorial synthesis and sensorial properties of 21 mercapto esters. AB - The aim of this work was to determine the chromatographic and sensorial properties of 21 synthetic mercapto acetates, among which two-3-mercaptohexyl acetate and 3-methyl-3-mercaptobutyl-acetate-have been previously detected in foods, especially passion fruit and wine. Combinatorial chemistry was used to synthesize them from alpha,beta-unsaturated ketones and aldehydes in a very short time. GC-SCD, GC-MS, and GC-olfactometry proved to be very helpful techniques for analyzing them individually in the reaction mixture obtained. Compared to other polyfunctional thiols, mercapto esters show relatively high BE-GC-LoADS values (best estimate-gas chromatography-lowest amount detected by sniffing), with very diverse descriptors. PMID- 12769536 TI - Combinatorial synthesis and sensorial properties of mercapto primary alcohols and analogues. AB - Combinatorial chemistry was used to extend the knowledge of beer thiols by synthesizing 13 mercapto primary alcohols and 6 other compounds (mercapto ketones and corresponding alcohols). Their respective mass spectra, retention indices on two columns, odor descriptors, and best estimated-gas chromatography-lowest amounts detected sniffing (BE-GC-LoADS) are reported. The descriptor "cheese", commonly associated with mercapto aldehydes and thioesters, was rarely used for mercapto alcohols or mercapto ketones. Polyfunctional thiols containing at least six carbon atoms usually gave a delicate odor of rhubarb/carrot, whereas most of the mercapto primary alcohols of intermediate size were described as onion-like, plastic-like, or pungent. Sensorial analysis of beers spiked with copper(II) ions allowed confirmation of the participation of thiols in the overall odor of fresh beer. The finding of 4-mercapto-4-methyl-2-pentanone and 3-mercaptohexanol in an XAD-2 Amberlite resin beer extract indicates a requirement for a more efficient thiol extraction procedure allowing identification of relevant polyfunctional thiols for brewing scientists. PMID- 12769537 TI - Generation of thiols by biotransformation of cysteine-aldehyde conjugates with baker's yeast. AB - Baker's yeast was shown to catalyze the transformation of cysteine-furfural conjugate into 2-furfurylthiol. The biotransformation's yield and kinetics were influenced by the reaction parameters such as pH, incubation mode (aerobic and anaerobic), and substrate concentration. 2-Furfurylthiol was obtained in an optimal 37% yield when cysteine-furfural conjugate at a 20 mM concentration was anaerobically incubated with whole cell baker's yeast at pH 8.0 and 30 degrees C. Similarly to 2-furfurylthiol, 5-methyl-2-furfurylthiol (11%), benzylthiol (8%), 2 thiophenemethanethiol (22%), 3-methyl-2-thiophenemethanethiol (3%), and 2 pyrrolemethanethiol (6%) were obtained from the corresponding cysteine-aldehyde conjugates by incubation with baker's yeast. This work indicates the versatile bioconversion capacity of baker's yeast for the generation of thiols from cysteine-aldehyde conjugates. Thanks to its food-grade character, baker's yeast provides a biochemical tool to produce thiols, which can be used as flavorings in foods and beverages. PMID- 12769539 TI - Formation of odorants in Maillard model systems based on l-proline as affected by pH. AB - Formation of the odorants acetic acid, 4-hydroxy-2,5-dimethyl-3-(2H)-furanone (HDMF), 6-acetyl-1,2,3,4-tetrahydropyridine (ATHP), and 2-acetyl-1-pyrroline (AP) was monitored by isotope dilution assays at pH 6, 7, and 8 in Maillard model reactions containing glucose and proline (Glc/Pro) or the corresponding Amadori compound fructosyl-proline (Fru-Pro). In general, higher yields were obtained at pH 7 and 8. Acetic acid was the major odorant with up to 40 mg/mmol precursor followed by HDMF (up to 0.25 mg/mmol), the formation of which was favored in the Fru-Pro reaction systems. On the contrary, ATHP (up to 50 microg/mmol) and AP (up to 5 microg/mmol) were more abundant in Glc/Pro. However, the sensory relevance of the two N-heterocycles was more pronounced on the basis of odor activity values, confirming their contribution to the overall roasty note of the reaction samples. It was also found that formation and decomposition of Fru-Pro were faster at pH 7 as compared to pH 6, explaining in part the preferred formation of the four odorants studied under neutral and slightly alkaline conditions. After 4 h of reaction at pH 7 in the presence of proline, about one-fourth of the glucose was consumed leading to acetic acid with a transformation yield of almost 40 mol %. PMID- 12769538 TI - Comparison of nosespace, headspace, and sensory intensity ratings for the evaluation of flavor absorption by fat. AB - The goal of this study was to better understand the correspondence between sensory perception and in-nose compound concentration. Five aroma compounds at three different concentrations increasing by factors of 4 were added to four matrixes (water, skim milk, 2.7% fat milk, and 3.8% fat milk). These were evaluated by nosespace analysis with detection by proton transfer reaction mass spectrometry (PTR-MS), using five panelists. These same panelists evaluated the perceived intensity of each compound in the matrixes at the three concentrations. PTR-MS quantification found that the percent released from an aqueous solution swallowed immediately was between 0.1 and 0.6%, depending on the compound. The nosespace and sensory results showed the expected effect of fat on release, where lipophilic compounds showed reductions in release as fat content increases. The effect is less than that observed in headspace studies. A general correlation between nosespace concentration and sensory intensity ratings was found. However, examples of perceptual masking were found where higher fat milks showed reductions in aroma compound intensity ratings, even if the nosespace concentrations were the same. PMID- 12769540 TI - A kinetic model for the glucose-fructose-glycine browning reaction. AB - The browning of glucose-fructose-glycine mixtures involves parallel glucose glycine and fructose-glycine reactions, which share a common intermediate, the immediate precursor of melanoidins in the kinetic model. At pH 5.5, 55 degrees C glucose is converted into this intermediate in a two step process where k(1) = (7.8 +/- 1.1) x 10(-)(4) mol L(-)(1) h(-)(1) and k(2) = (1.84 +/- 0.31) x 10( )(3) h(-)(1) according to established kinetics, whereas fructose is converted into this intermediate in a single step where k(4) = 5.32 x 10(-)(5)()()mol L( )(1) h(-)(1). The intermediate is converted to melanoidins in a single rate limiting process where k(mix) = 0.0177 h(-)(1) and the molar extinction coefficient (based on the concentration of sugar converted) of the melanoidins so formed is 1073 +/- 4 mol(-)(1) L cm(-)(1). Whereas the value of k(mix) is the same when the individual sugars undergo browning, the value of the molar extinction coefficient is similar to that for melanoidins from the glucose glycine reaction (955 +/- 45 mol(-)(1) L cm(-)(1)) but it is approximately double the value for melanoidins from the fructose-glycine reaction (478 +/- 18 mol( )(1) L cm(-)(1)). This is the reason that the effects of glucose and fructose on the rate of browning are synergistic. PMID- 12769541 TI - Antiproliferative and chemopreventive effects of adlay seed on lung cancer in vitro and in vivo. AB - This study examined the effects of different extracts of adlay seed on the growth of human lung cancer cells in vitro and in vivo. The data showed that a methanolic extract, but not a water extract, of adlay seed exerted an antiproliferative effect on A549 lung cancer cells by inducing cell cycle arrest and apoptosis. It was also found that tumor growth in vivo was inhibited by the methanolic extract in a dose-dependent manner. The chemopreventive effect of adlay seed on the tobacco-specific carcinogen 4-(methylnitrosamino)-1-(3-pyridyl) 1-butanone (NNK)-induced lung tumorigenesis in A/J mice was also investigated. Groups of mice were pre-fed with different diets, followed by feeding with NNK containing drinking water for 8 months. The results indicated that feeding with diet containing 30% of powdered adlay seed reduced the number of surface lung tumors by approximately 50%. Taken together, these results indicate that the components of adlay seed exert an anticancer effect in vitro and in vivo and may be useful for the prevention of lung tumorigenesis. PMID- 12769542 TI - Antioxidant activity of peptides obtained from porcine myofibrillar proteins by protease treatment. AB - Hydrolysates obtained from porcine myofibrillar proteins by protease treatment (papain or actinase E) exhibited high antioxidant activity in a linolenic acid peroxidation system induced by Fe(2+). Hydrolysates produced by both papain and actinase E showed higher activities at pH 7.1 than at pH 5.4. The antioxidant activity of the papain hydrolysate was almost the same as that of vitamin E at pH 7.0. These hydrolysates possessed 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity and chelating activity toward metal ions. Antioxidant peptides were separated from the papain hydrolysate by ion exchange chromatography. The acidic fraction obtained by this method exhibited higher activity than the neutral or basic fractions. Antioxidant peptides in the acidic fraction were isolated by high-performance liquid chromatography on an ODS column and shown to possess the structures DSGVT, IEAEGE, DAQEKLE, EELDNALN, and VPSIDDQEELM. The DAQEKLE peptide showed the highest activity among these peptides. PMID- 12769543 TI - Antioxidative properties of tripeptide libraries prepared by the combinatorial chemistry. AB - Two series of combinatorial tripeptide libraries were constructed, based on an antioxidative peptide isolated from a soybean protein hydrolysate. One was a library of 108 peptides containing either His or Tyr residues. Another was a library of 114 peptides related to Pro-His-His, which had been identified as an active core of the antioxidative peptide. The antioxidative properties of these libraries were examined by several methods, such as the antioxidative activity against the peroxidation of linoleic acid, the reducing activity, the radical scavenging activity, and the peroxynitrite scavenging activity. Two Tyr-containg tripeptides showed higher activities than those of two His-containing tripeptides in the peroxidation of linoleic acid. Tyr-His-Tyr showed a strong synergistic effects with phenolic antioxidants. However, the tripeptide had only marginal reducing activity and a moderate peroxynitrite scavenging activity. Cysteine containing tripeptides showed the strong peroxynitrite scavenging activity. Change of either the N-terminus or C-terminus of Pro-His-His to other amino acid residues did not significantly alter their antioxidative activity. Tripeptides containing Trp or Tyr residues at the C-terminus had strong radical scavenging activities, but very weak peroxynitrite scavenging activity. The present results allow us to understand why protein digests have such a variety of antioxidative properties. PMID- 12769544 TI - From plums to prunes: influence of drying parameters on polyphenols and antioxidant activity. AB - Prunes, which are industrially obtained by dehydrating fresh plums at 85-90 degrees C for 18 h, contain higher levels of phenolic compounds than most other fruits. Prune phenolics have shown beneficial effects on human health. Reports are available in the literature on ascorbic acid, phenol composition, and antioxidant activity of fresh plums and prunes, but there is a lack of publications on the influence of drying parameters on the phenolic compounds and antioxidant activity. A study was carried out on two plum cultivars using two sets of air-drying temperatures: (i) air temperature at 85 degrees C until 50% of prune moisture level and then the temperature was lowered to 70 degrees C; (ii) air temperature at 60 degrees C. Whole fresh and dried fruits were assessed for phenolics (catechins, hydroxycinnamic acids, anthocyanins, and flavonols), ascorbic acid, and antioxidant activity (all parameters were calculated on a dry matter basis). Analysis of data shows that chlorogenic and neochlorogenic acid changes were affected by both process parameters and cultivar. Drying destroyed anthocyanins, and there was a significant decrease in flavonols. Ascorbic acid was drastically reduced in relation to process temperature. The most striking result was that drying at 85 degrees C doubled antioxidant activity in both cultivars, while contradictory results were found for 60 degrees C processed plums. PMID- 12769545 TI - Pathway leading to the formation of anthocyanin-vinylphenol adducts and related pigments in red wines. AB - On the basis of observations from Vitis vinifera cv. Pinotage wines and experiments performed in model wine medium, a new chemical pathway responsible for the formation of anthocyanin-vinylphenol adducts in red wines is described. Until now, these pigments have been considered to be reaction products of anthocyanins and vinylphenols, the latter being generated during fermentation by enzymatic decarboxylation of the respective cinnamic acids. The mechanism of the novel pathway, involving intact hydroxycinnamic acid and anthocyanin, is explained. Only cinnamic acids with electron-donating substituents on the aromatic ring, such as coumaric acid, ferulic acid, caffeic acid, and sinapic acid, undergo this conversion, as they stabilize an intermediately formed carbenium ion. Decarboxylation and oxidation of the pyran moieties are the final steps in the generation of the corresponding 4-vinylphenol, 4-vinylguaiacol, 4 vinylcatechol, and 4-vinylsyringol adducts of anthocyanins in red wine. PMID- 12769547 TI - The oxidation of diamond: the geometry and stretching frequency of carbonyl on the (100) surface. AB - Microwave plasma deposited (100) diamond films have been thermally oxidized in dry O2 between 500 and 723 degrees C. The roughness of a single crystalline grain following oxidation is consistent with a layer-by-layer mechanism for the removal of carbon monoxide. The resulting surface exhibits infrared absorption bands at 1731 and 905 cm-1, attributed to the stretching and bending modes of a surface bonded carbonyl group. The former is within 1 cm-1 of the structurally analogous molecule 2-adamantanone. These data are consistent with the carbonyl groups being present on diamond (100) terraces. PMID- 12769546 TI - Assessment of concentrations of iron and zinc and bioavailable iron in grains of early-maturing tropical maize varieties. AB - Twenty elite early-maturing (75-90 days) tropical maize varieties grown in three diverse agroecologies in West Africa were evaluated to identify varieties with high kernel-Fe and -Zn and bioavailable Fe levels. Bioavailable iron was assessed using an in vitro digestion/Caco-2 cell model. Significant (P < 0.001) varietal differences were observed in mean kernel-Fe and -Zn levels. The ranges were 15.5 19.1 mg kg(-)(1) for Fe and 16.5-20.5 mg kg(-)(1) for Zn. Genetic component accounted for 34% of the total variation in kernel-Zn and for 11% of the variation in kernel-Fe levels. Mean bioavailable Fe in varieties ranged between 4% below and 49% above the reference control variety. A significant negative relationship was detected between kernel-P concentration and bioavailable Fe (R = -0.36; P < 0.004; n = 60). Two varieties, ACR90POOL16-DT and ACR86TZESR-W, were identified as the most promising for further evaluation to determine their efficacy as improved sources of iron in target populations. PMID- 12769548 TI - Optically active polyelectrolyte multilayers as membranes for chiral separations. AB - Ultrathin films of chiral polyelectrolyte complex, prepared by the multilayering process, exhibit selectivity in the membrane separations of optically active compounds, such as l- and d-ascorbic acid. The flux through these polyelectrolyte multilayers, PEMUs, is exceptionally high and may be controlled by the concentration of salt present in the permeating solutions. Both in-situ ATR-FTIR and chiral capillary electrochromatography indicate that flux selectivity is mainly kinetically controlled, stemming from a difference in diffusion rates of various enantiomers through PEMUs, rather than a difference in partitioning. PMID- 12769549 TI - Oscillating formation of 8-oxoguanine during DNA oxidation. AB - Oxidation of free guanine and guanine in salmon testes ds-DNA by hydroxyl radicals generated with Fenton reagent resulted in oscillating 8-oxoguanine concentrations. These oscillations were superimposed on a general trend of decreasing ratio of [8-oxoguanine]/{[8-oxoguanine] + [guanine]} with time, suggesting that a steady state 8-oxoguanine concentration would not be achieved. Mass spectrometry detected 8-oxoguanine and 5-guanidinohydantoin as products, suggesting that the latter was the product of oxidation of 8-oxoguanine. Guanidinohydantoin and other possible intermediates and products may be involved in a complex mechanism leading to the observed behavior. Oscillatory fluctuations in 8-oxoguanine may need to be considered in assessing its clinical significance as a biomarker for oxidative DNA damage. PMID- 12769550 TI - Constraints on supramolecular structure in amyloid fibrils from two-dimensional solid-state NMR spectroscopy with uniform isotopic labeling. AB - We show that strong constraints on supramolecular structure in amyloid fibrils can be obtained from solid-state nuclear magnetic resonance measurements on samples with uniformly 13C-labeled segments. The measurements exploit two dimensional (2D) 13C-13C exchange spectroscopy in conjunction with high-speed magic angle spinning, with proton-mediated exchange of 13C nuclear spin magnetization as recently demonstrated by Baldus and co-workers (J. Am. Chem. Soc. 2002, 124, 9704-9705). Proton-mediated 2D exchange spectra of fibrils formed by residues 16-22 of the 40-residue Alzheimer's beta-amyloid peptide show strong nonsequential, intermolecular cross-peaks between alpha-carbons that dictate an antiparallel beta-sheet structure in which residue 16+k aligns with residue 22-k. The strong alpha/alpha cross-peaks are absent from conventional, direct 2D exchange spectra. Proton-mediated 2D exchange spectra of fibrils formed by residues 11-25 indicate an antiparallel beta-sheet structure with a pH-dependent intermolecular alignment. In contrast, proton-mediated 2D exchange spectra of fibrils formed by the full-length beta-amyloid peptide are consistent with a parallel beta-sheet structure. These data show that the supramolecular structure of amyloid fibrils is not determined by the amino acid sequence at the level of 7 residue or 15-residue segments. The proton-mediated 2D exchange spectra additionally demonstrate that the intermolecular alignment in the beta-sheets of these amyloid fibrils is highly ordered, with no detectable evidence for "misalignment" defects. PMID- 12769551 TI - Hydrogen bonding as a construction element for bidentate donor ligands in homogeneous catalysis: regioselective hydroformylation of terminal alkenes. AB - A new concept for the construction of bidentate ligands for homogeneous metal complex catalysis is described. The concept relies on the self-assembly of monodentate ligands through hydrogen bonding. As a prototype of such systems, 6 diphenylphosphanyl-2-pyridone (6-DPPon) was shown to form a chelate in the coordination sphere of a transition metal center through unusual pyridone/hydroxypyridine hydrogen bonding (X-ray). This hydrogen bonding stays intact in a catalytic reaction as proven upon highly regioselective hydroformylation of terminal alkenes. Regioselectivities and reactivities observed rank the 6-DPPon/rhodium system among the most active and regioselective catalysts for n-selective hydroformylation of terminal alkenes. PMID- 12769552 TI - Chiral sulfoxides as neutral coordinate-organocatalysts in asymmetric allylation of N-acylhydrazones using allyltrichlorosilanes. AB - Sulfoxides were first introduced to the allylation of N-acylhydrazones with allyltrichlorosilanes as effective neutral coordinate-organocatalysts (NCOs). Both high diastereo- and enantioselectivity were attained when optically active chiral sulfoxides were used. Asymmetric crotylations using (Z)- and (E) crotyltrichlorosilanes showed a high level of stereospecificity (Z --> anti and E --> syn) with high enantioselectivity. PMID- 12769553 TI - Dynamic glycosylation of the transcription factor CREB: a potential role in gene regulation. AB - We report that CREB (cyclic AMP-responsive element-binding protein), a transcription factor essential for long-term memory, is O-GlcNAc glycosylated in the mammalian brain. Glycosylation occurs at two sites within the Q2 domain and disrupts the interaction between CREB and TAFII130, thereby repressing the transcriptional activity of CREB in vitro. These findings have important implications for the role of O-GlcNAc glycosylation in gene regulation, and they provide a link between O-GlcNAc and information storage processes in the brain. PMID- 12769554 TI - Quantum mechanical models correlating structure with selectivity: predicting the enantioselectivity of beta-amino alcohol catalysts in aldehyde alkylation. AB - Quantitative structure selectivity relationship (QSSR) models are described that provide consistently reliable predictions for the asymmetric addition of Et2Zn to PhCHO catalyzed by beta-amino alcohols. Statistically valid two-variable linear regression models that correlate the structures of the chiral catalysts with their enantioselectivities are obtained from three-dimensional physical property grids. The strength of the present method is that statistical models obtained from a small set of experimentally determined selectivities and relatively simple theoretical calculations yield selectivity predictions that are as accurate as those derived from higher-level calculations of transition-structure energies. Only minutes of computing time are required. Simple models are obtained which permit straightforward physical interpretation and generate realistic predictions. PMID- 12769555 TI - Enhanced D-amino acid incorporation into protein by modified ribosomes. AB - By overexpression of modified Escherichia coli 23S rRNAs from multicopy plasmids, ribosomes were prepared that contained mutations in two regions (2447-2450 and 2457-2462) of 23S rRNA. Following mutagenesis and selection, two clones with mutations in the 2447-2450 region (peptidyltransferase center) and six with mutations in the 2457-2462 region (helix 89) were characterized. The mutations were shown to exhibit a high level of homology. Cell-free protein synthesizing systems prepared from these clones were found to exhibit significantly enhanced incorporation of d-methionine and d-phenylalanine into protein. The incorporations involved positions 10, 22, and 54 of E. coli dihydrofolate reductase and positions 247 and 250 of Photinus pyralis firefly luciferase. Interestingly, some of the derived proteins containing the d-amino acids (notably DHFR analogues altered at position 10) functioned as well as those containing the respective l-amino acids, while substitution at other positions resulted in proteins having greatly diminished activity. PMID- 12769556 TI - Reactivity of chalcogenostannate compounds: syntheses, crystal structures, and electronic properties of novel compounds containing discrete ternary anions [MII4(mu4-Se)(SnSe4)4]10- (MII = Zn, Mn). AB - Reactions of K4[SnSe4].1.5MeOH with ZnCl2 or MnCl2.4H2O in water/methanol mixtures yield novel compounds [K10(H2O)16(MeOH)0.5][M4(mu4-Se)(SnSe4)4] (M = Zn, 2; Mn, 3) in high yields; 2 and 3 contain the first discrete ternary Zn/Sn/Se or Mn/Sn/Se cluster anions. Both compounds were unambiguously characterized by X-ray diffraction (tetragonal, space groups P43212 and P41212, respectively) revealing chiral anionic structures within chiral crystals. Optical spectra of 2 and 3 indicate energy differences for the lowest electronic excitations (Eg = 2.57 eV, 2; 2.27 eV, 3) that are very close to the band gap values observed for mesoporous solids with polymeric M/Sn/E networks. DFT investigations on the electronic situation and first ESR studies agree in that they demonstrate a high-spin ground state in the case of 3 with 20 unpaired electrons at four uncoupled MnII centers. PMID- 12769557 TI - A novel biaxial smectic liquid crystalline phase formed by rodlike molecules with a 1,3-diazaazulene skeleton. AB - Novel rodlike liquid crystalline (LC) materials incorporating the nonbenzenoid aromatic 1,3-diazaazulene unit have been synthesized and investigated by polarizing microscopy, differential scanning calorimetry, and X-ray diffraction of oriented samples. Some representatives form, in addition to conventional SmC phases, a novel biaxial smectic LC phase, in which the rotation of the molecules around their long axes is greatly restricted. Attractive face-to-face interactions of the flat aromatic units lead to columnar aggregates which are organized in layers. As there is no positional correlation between adjacent layers, these mesophases are related to lamellar columnar phases formed by some disklike and boardlike molecules. Such LC materials may be promising candidates in the field of organic semiconductors, photoconduction, and other applications. PMID- 12769558 TI - Chemical formation of mononuclear univalent zinc in a microporous crystalline silicoaluminophosphate. AB - A host-guest compound containing mononuclear univalent Zn+ ions has been prepared through vapor reaction of metallic zinc with protons in the cages of SAPO-CHA, a silicoaluminophosphate molecular sieve. Unlike in fused ZnCl2 salt, the formed Zn+ species are associated with the molecular sieve walls, and this prevents the cations from getting paired to form diamagnetic (Zn-Zn)2+. Electron spin resonance spectroscopy reveals the paramagnetic nature of the Zn+ species which display anisotropic g factors, and the two crystallographically independent Zn+ sites are distinguishable from the ESR signals. The magnetic moments of the univalent zinc in SAPO-CHA interact antiferromagnetically at low temperatures with a Neel point of about 4 K as observed from the temperature dependence plot of the magnetic susceptibility of the host-guest compound. PMID- 12769559 TI - Rhodium-catalyzed disulfide exchange reaction. AB - A system of RhH(PPh3)4, trifluoromethanesulfonic acid, and (p-tol)3P catalyzes the disulfide exchange reaction. Treatment of two symmetrical dialkyl disulfides with the catalyst provides an equilibrium mixture of three disulfides within 15 min in refluxing acetone. The catalyst is active after reaching the equilibrium, and addition of a disulfide to the mixture changes the ratio of the products. The use of 4 mol equiv excess of one of the disulfides provides the unsymmetrical disulfide in a yield exceeding 80%. Disulfide-containing peptides also undergo an exchange reaction. The reactions of diaryl disulfides and dialkyl disulfides are even faster, and reach equilibrium within 5 min at room temperature in the presence of the rhodium complex and 1,2-bis(diphenylphosphino)ethane (dppe). This exchange reaction is considerably affected by the substituents on the disulfides. Treatment of diphenyl disulfide, di(p-tolyl) disulfide, and bis(sec-butyl) disulfide yields phenyl p-tolyl disulfide at room temperature with unchanged bis(sec-butyl) disulfide; random disproportionation occurs at reflux. The rhodium catalysis can be used for the exchange reaction of disulfides and diselenides giving selenosulfides as well as disulfides and ditellurides giving tellurinosulfides. PMID- 12769560 TI - Probing hydrogen bonding in a DNA triple helix using protium-deuterium fractionation factors. AB - In this communication we report protium-deuterium fractionation factors for the intramolecular triple helix formed by the DNA oligonucleotide 5' d(AGAGAGAACCCCTTCTCTCTTTTTCTCTCTT)-3'. The fractionation factors of individual Watson-Crick and Hoogsteen hydrogen bonds in the structure are measured by NMR spectroscopy. The results show that, in contrast to proteins, the fractionation factors are all equal or lower than unity. On the average, the values of the fractionation factors are centered between 0.6 and 0.8, and no significant differences are observed between Hoogsteen and Watson-Crick hydrogen bonds. Deviations from the average are observed for the 5'-end region of the molecule where a base triad is absent and the structure is strained by the intramolecular folding of the DNA strand. PMID- 12769561 TI - Secondary losses via gamma-lactam formation in hot electron capture dissociation: a missing link to complete de novo sequencing of proteins? AB - Differentiating between the isomeric isoleucine and leucine amino acid residues can be accomplished by a gas-phase fragmentation reaction in which an N-terminal radical site of a C-terminal fragment initiates formation of a five-membered ring with the beta-carbon in the adjacent residue. PMID- 12769562 TI - Total synthesis of the duocarmycins. AB - The total synthesis of (+)-duocarmycin A and SA through a common indoline intermediate is described. The key reactions include selective lithiation of a 2,6-dibromoiodobenzene derivative and diastereoselective addition to a chiral nitroalkene, copper-mediated aryl amination, and addition of aryllithium to azlactones. PMID- 12769563 TI - Controlling the aqueous miscibility of ionic liquids: aqueous biphasic systems of water-miscible ionic liquids and water-structuring salts for recycle, metathesis, and separations. AB - Hydrophilic ionic liquids can be salted-out and concentrated from aqueous solution upon addition of kosmotropic salts forming aqueous biphasic systems as illustrated by the phase behavior of mixtures of 1-butyl-3-methylimidazolium chloride ([C4mim]Cl) and K3PO4. PMID- 12769565 TI - Thermodynamic contributions of the ordered water molecule in HIV-1 protease. AB - Binding between biomolecules is usually accompanied by the formation of direct interactions with displacement of water from the binding sites. In some cases, however, the interactions are mediated by ordered water molecules, whose effect on binding affinity and the other thermodynamic functions is unclear. In this work, we compute the contribution of one such water molecule, the strongly bound water molecule at the binding site of HIV-1 protease, to the thermodynamic properties using statistical mechanical formulas for the energy and entropy. The requisite correlation functions are obtained by molecular dynamics simulations. We find that the entropic penalty of ordering is large but is outweighed by the favorable water-protein interactions. We also find a large negative contribution from this water molecule to the heat capacity. This approach could be useful in rational drug design by estimating which bound water molecules would be most favorable to displace. PMID- 12769566 TI - Palladium-catalyzed bissilylation of arynes with cyclic disilanes: synthesis of benzo-annulated disilacarbocycles. AB - Arynes, generated in situ from 2-(trimethylsilyl)aryl triflate and a fluoride ion, were found to insert into a silicon-silicon bond of cyclic disilanes in the presence of a catalytic amount of a palladium-tert-alkyl isocyanide complex to afford diverse benzo-annulated disilacarbocycles straightforwardly in modest to high yields. PMID- 12769564 TI - EDTA-derivatized deoxythymidine as a tool for rapid determination of protein binding polarity to DNA by intermolecular paramagnetic relaxation enhancement. AB - EDTA-derivatized deoxythymidine (dT-EDTA), incorporated into DNA and complexed to Fe2+ in the presence of dithiothreitol, is a widely used reagent for sequence specific cleavage of duplex DNA. Using HPLC/electrospray mass spectrometry, we show that cleavage is specific to Fe2+, and no cleavage occurs when DNA-EDTA is complexed to other metal ions such as Ca2+, Mn2+, and Fe3+ even after many days. Because dT-EDTA can be incorporated at any desired position of a synthetic oligonucleotide, DNA-EDTA is ideally suited for the measurement of intermolecular paramagnetic relaxation enhancement effects between a paramagnetic ion chelated to DNA-EDTA and a bound protein. Measurements on the SRY/DNA-EDTA complex using two double-stranded oligonucleotides bearing dT-EDTA at opposite ends of the sequence indicate that intermolecular 1HN-T2 enhancement by chelated Mn2+ can be used to readily ascertain the polarity of protein binding to DNA and to derive quantitative long-range distance information for structure refinement. In the case of the SRY-DNA complex, excellent agreement between observed and calculated 1HN-T2 paramagnetic relaxation enhancement data can be achieved with insignificant shifts in the atomic coordinates ( approximately 0.25 A for all heavy atoms) while simultaneously satisfying all other experimental restraints. A unique feature of DNA-EDTA is that the relaxation enhancement effect can be tuned by judicious choice of the paramagnetic metal ion, thereby permitting a wide range of long-range intermolecular electron-proton distances, ranging from approximately 9 to 35 A, to be probed. PMID- 12769567 TI - Rapid radical formation by DNA charge transport through sequences lacking intervening guanines. AB - Using the flash-quench technique to probe DNA charge transport in assemblies containing a tethered ruthenium intercalator, the kinetics and yield of methylindole radical formation as a function of DNA sequence were studied by laser spectroscopy and biochemical methods. In these assemblies, the methylindole moiety serves as an artificial base of low oxidation potential. Hole injection and subsequent formation of the methylindole radical cation were observed at a distance of over 30 A at rates >/=107 s-1 in assemblies containing no guanine bases intervening the ruthenium intercalator and GMG oxidation site. Radical yield was, however, strikingly sensitive to an intervening base mismatch; no significant methylindole radical formation was evident with an intervening AA mismatch. Also critical is the sequence at the injection site; this sequence determines initial hole localization and hence the probability of hole propagation. With guanine rather than inosine near the site of hole injection, decreased yields of radicals and long-range oxidative damage are observed. The presence of the low-energy guanine site in this case serves to localize the hole and therefore diminish charge transport through the base pair stack. PMID- 12769568 TI - A colorimetric lead biosensor using DNAzyme-directed assembly of gold nanoparticles. AB - A highly sensitive and selective colorimetric lead biosensor based on DNAzyme directed assembly of gold nanoparticles is reported. It consists of a DNAzyme and its substrate that can hybridize to a 5'-thio-modified DNA attached to gold nanoparticles. The hybridization brings gold nanoparticles together, resulting in a blue-colored nanoparticle assembly. In the presence of lead, the DNAzyme catalyzes specific hydrolytic cleavage, which prevents the formation of the nanoparticle assembly, resulting in red-colored individual nanoparticles. The detection level can be tuned to several orders of magnitude, from 100 nM to over 200 muM, through addition of an inactive variant of the DNAzyme. The concept developed here can be applied to the design of nucleic acid enzyme/nanoparticle sensors for analytes that are subject to in vitro selection, and thus can significantly expand the scope of nanomaterial applications and provide a novel approach to designing simple colorimetric biosensors. PMID- 12769569 TI - Electrochemical resolution of 15 oxidation states for monolayer protected gold nanoparticles. AB - The first observation of 15 voltammetric quantized charging peaks for a solution of hexanethiol-capped gold nanoparticles (so-called monolayer protected clusters MPCs) at room temperature is reported where the variation in peak spacing with increasing charge stored in the metal core is discussed in terms of MPC capacitance. PMID- 12769570 TI - Carbon-carbon bond cleavage of diynes through the hydroamination with transition metal catalysts. AB - The C-C bond cleavage of terminal and internal diynes takes place readily in the presence of catalytic amounts of Ru3(CO)12 or Pd(NO3)2 and of 2-aminophenol, giving the corresponding benzoxazoles and ketones in good to high yields. There are two different modes of the bond cleavage: (a) an alkyne C-C triple bond is cleaved, and (b) the C-C single bond between the two alkyne groups is cleaved. PMID- 12769571 TI - Spectroscopic evidence for a heme-superoxide/Cu(I) intermediate in a functional model of cytochrome c oxidase. AB - A superstructured tetraphenylporphyrin with a covalently attached proximal imidazole axial base and three distal imidazole pickets has been developed as a model for the active site of terminal oxidases such as cytochrome c oxidase. The oxygen adduct of the Fe-only heme (at low temperature) has a diamagnetic NMR and is EPR silent, which taken together with a resonance Raman oxygen isotope sensitive band (nuFe-O) at 575/554 cm-1 (16O2/18O2) indicates formation of a six coordinate heme-superoxide complex. Unexpectedly, the Fe/Cu complex, where the copper is in a trisimidazole environment approximately 5 A above the heme plane, displays similar characteristics: a diamagnetic NMR, EPR silence, and nuFe-O at 570/544 cm-1. This indicates the dioxygen adduct of this Fe/Cu system is also a superoxide. This contrasts with previously characterized partially reduced dioxygen intermediates of binuclear heme/copper complexes that form Fe/Cu mu peroxo complexes. PMID- 12769572 TI - Enantioselective total synthesis of briarellins E and F: the first total syntheses of briarellin diterpenes. AB - Enantioselective total syntheses of briarellin E (4) and briarellin F (5) have been achieved starting with (S)-(+)-carvone and (S)-(-)-glycidol. These total syntheses are the first of briarellin diterpenes. The central step in these syntheses is acid-promoted condensation of cyclohexadienyl diol 15 and (Z) alpha,beta-unsaturated aldehyde 16 to form, with complete stereocontrol, the hexahydroisobenzofuran core and six stereocenters of these coral metabolites. These syntheses also feature stereospecific photolytic deformylation of beta,gamma-unsaturated aldehyde 17 to remove the extraneous carbon introduced in the Prins-pinacol step, chemo- and stereoselective hydroxyl-directed epoxidation of dienyl alcohol 18 to incorporate the C3 oxygen stereocenter, regio- and stereoselective rearrangement of epoxy ester 19 to install the C4 oxygen substituent, efficient dehydrative cyclization of a 1,6-diol intermediate to form the oxepane ring, and diastereoselective Nozaki-Hiyama-Kishi cyclization of vinyl iodide aldehyde 25 to forge the oxacyclononane ring and the C6 hydroxyl stereocenter. These total syntheses establish the absolute configurations of 4 and 5, define a concise strategy for the total synthesis of briarellin diterpenes, and provide additional illustrations of the uncommon utility of pinacol-terminated cationic cyclizations for stereocontrolled synthesis of complex oxacyclic natural products. PMID- 12769573 TI - Expanding Pd-catalyzed C-N bond-forming processes: the first amidation of aryl sulfonates, aqueous amination, and complementarity with Cu-catalyzed reactions. AB - The first general method for the Pd-catalyzed amination of aryl tosylates and benzenesulfonates was developed utilizing ligand 1, which belongs to a new generation of biaryl monophosphine ligands. In addition, the new catalyst system for the first time enables amidation of aryl arenesulfonates and aqueous amination protocols that do not necessitate the use of cosolvents. The substrate scope has been significantly expanded to include aryl halides containing primary amides and free carboxylic acid groups. In the case of multifunctional substrates, the Pd-catalyzed amination can provide selectivity that is complementary to the Cu-catalyzed C-N bond-forming processes. PMID- 12769574 TI - Sequence-selective DNA cleavage by a chimeric metallopeptide. AB - A chimeric metallopeptide derived from the sequences of two structurally superimposable motifs was designed as an artificial nuclease. Both DNA recognition and nuclease activity have been incorporated into a small peptide sequence. P3W, a 33-mer peptide comprising helices alpha2 and alpha3 from the engrailed homeodomain and the consensus EF-hand Ca-binding loop binds one equivalent of lanthanides or calcium and folds upon metal binding. The conditional formation constants (in the presence of 50 mM Tris) of P3W for Eu(III) (K(a) = (2.1 +/- 0.1) x 10(5) M(-1)) and Ce(IV) (K(a) = (2.6 +/- 0.1) x 10(5) M(-1)) are typical of isolated EF-hand peptides. Circular dichroism studies show that 1:1 CeP3W is 26% alpha-helical and EuP3W is up to 40% alpha-helical in the presence of excess metal. The predicted helicity of the folded peptide based on helix length and end effects is about 50%, showing the metallopeptides are significantly folded. EuP3W has considerably more secondary structure than our previously reported chimeras (Welch, J. T.; Sirish, M.; Lindstrom, K. M.; Franklin, S. J. Inorg. Chem. 2001, 40, 1982-1984). Eu(III)P3W and Ce(IV)P3W nick supercoiled DNA at pH 6.9, although EuP3W is more active at pH 8. CeP3W cleaves linearized, duplex DNA as well as supercoiled plasmid. The cleavage of a 5'-(32)P labeled 121-mer DNA fragment was followed by polyacrylamide gel electrophoresis. The cleavage products are 3'-OPO(3) termini exclusively, suggesting a regioselective or multistep mechanism. In contrast, uncomplexed Ce(IV) and Eu(III) ions produce both 3'-OPO(3) and 3'-OH, and no evidence of 4'-oxidative cleavage termini with either metal. The complementary 3'-(32)P-labeled oligonucleotide experiment also showed both 5'-OPO(3) and 5'-OH termini were produced by the free ions, whereas CeP3W produces only 5'-OPO(3) termini. In addition to apparent regioselectivity, the metallopeptides cut DNA with modest sequence discrimination, which suggests that the HTH motif binds DNA as a folded domain and thus cleaves selected sequences. The de novo artificial nuclease LnP3W represents the first small, underivatized peptide that is both active as a nuclease and sequence selective. PMID- 12769576 TI - Deoxyribozyme-based half-adder. AB - We have constructed a solution-phase array of three deoxyribozyme-based logic gates that behaves as a half-adder. Two deoxyribozymes mimic i(1)ANDNOTi(2) and i(2)ANDNOTi(1) gates that cleave a fluorogenic substrate, reporting through an increase in fluorescence emission at 570 nm. The third deoxyribozyme mimics an i(1)ANDi(2) gate and cleaves the other fluorogenic substrate, reporting through an increase in fluorescence emission at 520 nm. Together, this system represents the first example of a decision-making enzymatic network with two inputs and two outputs. Similar systems could be applied to control autonomous therapeutic and diagnostic devices. PMID- 12769575 TI - Investigation of solvent effects for the Claisen rearrangement of chorismate to prephenate: mechanistic interpretation via near attack conformations. AB - Solvent effects on the rate of the Claisen rearrangement of chorismate to prephenate have been examined in water and methanol. The preequilibrium free energy differences between diaxial and diequatorial conformers of chorismate, which had previously been implicated as the sole basis for the observed 100-fold rate increase in water over methanol, have been reframed using the near attack conformation (NAC) concept of Bruice and co-workers. Using a combined QM/MM Monte Carlo/free-energy perturbation (MC/FEP) method, 82%, 57%, and 1% of chorismate conformers were found to be NAC structures (NACs) in water, methanol, and the gas phase, respectively. As a consequence, the conversion of non-NACs to NACs provides no free-energy contributions to the overall relative reaction rates in water versus methanol. Free-energy perturbation calculations yielded differences in free energies of activation for the two polar protic solvents and the gas phase. The rate enhancement in water over the gas phase arises from preferential hydration of the transition state (TS) relative to the reactants via increased hydrogen bonding and long-range electrostatic interactions, which accompany bringing the two negatively charged carboxylates into closer proximity. More specifically, there is an increase of 1.3 and 0.6 hydrogen bonds to the carboxylate groups and the ether oxygen, respectively, in going from the reactant to the TS in water. In methanol, the corresponding changes in hydrogen bonding with first shell solvent molecules are small; the rate enhancement arises primarily from the enhanced long-range interactions with solvent molecules. Thus, the reaction occurs faster in water than in methanol due to greater stabilization of the TS in water by specific interactions with first shell solvent molecules. PMID- 12769578 TI - Attachment of styrene and phenylacetylene on Si(111)-7 x 7: the influence of substitution groups on the reaction mechanism and formation of pi-conjugated skeletons. AB - The interactions of styrene and phenylacetylene and their isotope substitutions with a Si(111)-7 x 7 surface have been studied as model systems to mechanistically understand the chemical binding of conjugated pi-electron systems to di-radical-like silicon dangling bonds of the adjacent adatom-rest atom pair. Vibrational studies show that styrene mainly binds to the surface through a diradical reaction involving both the external C=C and its conjugated internal C=C of the phenyl ring with an adjacent adatom-rest atom pair, forming a 5 ethylidene-1,3-cyclohexadiene-like skeleton. On the other hand, phenylacetylene was shown to be covalently attached to Si(111)-7 x 7 through the external C[triple bond]C, forming a styrene-like conjugation system. These experimental results are consistent with density functional theory calculations. The different binding mechanisms for styrene and phenylacetylene clearly demonstrate that reaction channels for multifunctional organic molecules are strongly dependent on the chemical and physical properties of the functional groups. The resulting pi electron conjugation structures may possibly be employed as intermediates for further organic syntheses and fabrication of multilayer organic films on semiconductor surfaces. PMID- 12769577 TI - Efficient macrocyclization of U-turn preorganized peptidomimetics: the role of intramolecular H-bond and solvophobic effects. AB - Simple peptidomimetic molecules derived from amino acids were reacted with meta- and para-bis(bromomethyl)benzene in acetonitrile to very efficiently yield macrocyclic structures. The cyclization reaction does not require high dilution techniques and seems to be insensitive to the size of the formed macrocycle. The analysis of data obtained by (1)H NMR, single-crystal X-ray diffraction, fluorescence measurements, and molecular mechanics indicate that folded conformations can preorganize the system for an efficient cyclization. The role played by intramolecular hydrogen-bonding and solvophobic effects in the presence of folded conformations is analyzed. PMID- 12769579 TI - Synthesis of functional olefin copolymers with controllable topologies using a chain-walking catalyst. AB - The branching topology of ethylene polar copolymers was for the first time successfully controlled by copolymerization of ethylene with polar olefins using a palladium-bisimine chain-walking catalyst, in which ethylene pressure and comonomer concentration were used to control the competition between isomerization (chain-walking) and monomer insertion processes. Although the overall branching density changes very slightly, the topology of the copolymers becomes more dendritic as the ethylene pressure and comonomer feed concentration are decreasing. This provides a straightforward one-pot synthesis to access a full range of functional copolymers having controllable branching topologies. To demonstrate the utility of this methodology, dendritic functional copolymers having hydroxyl, epoxide, and carbohydrate groups were prepared in a one-pot polymerization as potential functional materials. PMID- 12769580 TI - Effect of chromophore-charge distance on the energy transfer properties of water soluble conjugated oligomers. AB - The synthesis of 1,4-bis(9,9'-bis(3"-(N,N,N-trimethylammonium)-propyl)-2' fluorenyl)benzene tetrabromide (C3), 1,4-bis(9,9'-bis(4"-(N,N,N trimethylammonium)-butyl)-2'-fluorenyl)benzene tetrabromide (C4), 1,4-bis(9,9' bis(6"-(N,N,N-trimethylammonium)-hexyl)-2'-fluorenyl)benzene tetrabromide (C6), and 1,4-bis(9,9'-bis(8"-(N,N,N-trimethylammonium)-octyl)-2'-fluorenyl)benzene tetrabromide (C8) is reported. Fluorescence energy transfer experiments between C3-C8 and the acceptors pentasodium 1,4-bis(4'(2",4"-bis(butoxysulfonate) styryl)styryl)-2-(butoxysulfonate)-5-methoxybenzene (3), fluorescein labeled single-stranded DNA and fluorescein labeled double-stranded DNA in water, buffer, and methanol reveal the importance of hydrophobic and electrostatic forces in determining chromophore-chromophore close proximity. In water, the oligomers with longer side chain length show better energy transfer, as well as higher Stern Volmer quenching constants (K(sv)), largely due to a stronger hydrophobic attraction between the optically active components. In methanol, the differences in energy transfer are leveled, and the oligomers with shorter side chain lengths show higher K(sv) values. Compounds C3, C4, C6, and C8 were also used to dissect the different contributors to DNA hybridization assays based on cationic conjugated polymers. PMID- 12769581 TI - Dissociative and associative mechanisms of cope rearrangements of fluorinated 1,5 hexadienes and 2,2'-bis-methylenecyclopentanes. AB - The effects of fluorine substitution on the Cope rearrangements of 1,5-hexadiene and 2,2'-bis-methylenecyclopentane have been examined computationally using (U)B3LYP/6-31+G(d,p) and CASPT2/6-31G(d) methodology. The calculations indicate that fluorine substituents at the hexadiene termini generally stabilize the transition states relative to the ground states of the chair conformations and destabilize pathways that occur via boat conformations, in accord with the experimental observations of Dolbier. With meso-2,2' bis(difluoromethylene)cyclopentane, this destabilization is sufficient to favor a relatively dissociative concerted transition state resembling weakly interacting allyl radicals over a normal aromatic concerted boat transition state. This preference is due partly to an increase in the activation enthalpy for the concerted rearrangement coupled with the more favorable entropy for dissociation. In octafluoro- and decafluorohexadienes, the situation is reversed, and reaction through a cyclohexane-1,4-diyl is favored. Even in the octafluoro system with no radical stabilizing substituents at C(2) and C(5), the preference of fluorine for sp(3) centers causes reaction via the cyclohexane-1,4-diyl. In establishing methodology for this study, the conformations of 1,2-difluoroethane and 1,1,2,2 tetrafluoroethane were also examined thoroughly by the B3LYP method using three basis sets. PMID- 12769582 TI - Measuring molecular weight by atomic force microscopy. AB - Absolute-molecular-weight distribution of cylindrical brush molecules were determined using a combination of the Langmuir Blodget (LB) technique and Atomic Force Microscopy (AFM). The LB technique gives mass density of a monolayer, i.e., mass per unit area, whereas visualization of individual molecules by AFM enables accurate measurements of the molecular density, i.e., number of molecules per unit area. From the ratio of the mass density to the molecular density, one can determine the absolute value for the number average molecular weight. Assuming that the structure of brush molecules is uniform along the backbone, the length distribution should be virtually identical to the molecular weight distribution. Although we used only brush molecules for demonstration purpose, this approach can be applied for a large variety of molecular and colloidal species that can be visualized by a microscopic technique. PMID- 12769584 TI - Size-tunable emission from 1,3-diphenyl-5-(2-anthryl)-2-pyrazoline nanoparticles. AB - Organic nanoparticles of 1,3-diphenyl-5-(2-anthryl)-2- pyrazoline (DAP) ranging in average diameters from 40 to 160 nm were prepared through the reprecipitation method. The average diameters of the particles were controlled by variation of the aging time. We found that DAP nanoparticles exhibit the size-dependent optical properties. The absorption transitions of the nanoparticles at the lower energy side experience a bathochromic shift with an increase in the particle size as a result of the increased intermolecular interactions, while the higher-energy bands of anthracene split possibly due to the electronic coupling between the pyrazoline ring of one molecule and the anthracene moiety of the neighboring molecule. Most interestingly, the nanoparticle emission in the blue light region from pyrazoline chromophore shifts to shorter wavelengths with an increase in the particle size, accompanied with a relatively gradual dominance of the emission at about 540 nm from an exciplex between the pyrazoline ring of one molecule and the anthracene moiety of the neighboring molecule. The hypsochromic shift in the emission of DAP nanoparticles was identified as originating from the pronounced decrease in the Stokes shift due to the restraint of vibronic relaxation and the configuration reorganization induced by the increased intermolecular interaction. PMID- 12769583 TI - Characterization of a stable ruthenium complex with an oxyl radical. AB - The ruthenium oxyl radical complex, [Ru(II)(trpy)(Bu(2)SQ)O(.-)] (trpy = 2,2':6',2"-terpyridine, Bu(2)SQ = 3,5-di-tert-butyl-1,2-benzosemiquinone) was prepared for the first time by the double deprotonation of the aqua ligand of [Ru(III)(trpy)(Bu(2)SQ)(OH(2))](ClO(4))(2). [Ru(III)(trpy)(Bu(2)SQ)(OH(2))](ClO(4))(2) is reversibly converted to [Ru(III)(trpy)(Bu(2)SQ)(OH-)](+) upon dissociation of the aqua proton (pK(a) 5.5). Deprotonation of the hydroxo proton gave rise to intramolecular electron transfer from the resultant O(2-) to Ru-dioxolene. The resultant [Ru(II)(trpy)(Bu(2)SQ)O(.-)] showed antiferromagnetic behavior with a Ru(II) semiquinone moiety and oxyl radical, the latter of which was characterized by a spin trapping technique. The most characteristic structural feature of [Ru(II)(trpy)(Bu(2)SQ)O(.-)] is a long Ru-O bond length (2.042(6) A) as the first terminal metal-O bond with a single bond length. To elucidate the substituent effect of a quinone ligand, [Ru(III)(trpy)(4ClSQ)(OH(2))](ClO(4))(2) (4ClSQ = 4 chloro-1,2-benzosemiquinone) was prepared and we compared the deprotonation behavior of the aqua ligand with that of [Ru(III)(trpy)(Bu(2)SQ)(OH(2))](ClO(4))(2). Deprotonation of the aqua ligand of [Ru(III)(trpy)(4ClSQ)(OH(2))](ClO(4))(2) induced intramolecular electron transfer from OH- to the [Ru(III)(4ClSQ)] moiety affording [Ru(II)(trpy)(4ClSQ)(OH.)]+, which then probably changed to [Ru(II)(trpy)(4ClSQ)O(.-)]. The antiferromagnetic interactions (J values) between Ru(II)-semiquinone and the oxyl radical for [Ru(II)(trpy)(Bu(2)SQ)O(.-)] and for [Ru(II)(trpy)(4ClSQ)O(.-)] were 2J = -0.67 cm(-1) and -1.97 cm(-1), respectively. PMID- 12769585 TI - Towards graphite: magnetic properties of large polybenzenoid hydrocarbons. AB - The geometries of four different series of D(6h)-symmetric polybenzenoid hydrocarbons (PBH) up to and including C(222)H(42) have been optimized at the B3LYP/6-31G(d) level of theory. Excluding C(48)H(24) and C(138)H(42), which have D(3d) minima due to 1,5 H...H repulsions between adjacent perimeter rings, optimized geometries are planar D(6h) minima. Nucleus Independent Chemical Shifts (NICS), at the same level, indicate the presence of individual aromatic rings, which correspond to Clar's qualitative sextets rule (Clar, E. TheAromatic Sextet; Wiley: London, 1972). NICS and the Clar valence electron topologies agree perfectly in the molecule plane; however, the NICS values computed in parallel planes further away from the molecular surface converge, indicating the presence of a uniform magnetic shielding field. For each series, PBH total NICS values (i.e., the sum of NICS values for all rings in the PBH) correlate linearly with the number of carbon atoms, indicating constant magnetic field development within a series. The C-C lengths depend on their proximity to the more olefinic rich molecular perimeters. However, the large PBH (> or =C(48)H(24)) internal C-C distances converge to approximately 1.426 A. In agreement with Clar's rule, HF/6 31G(d)//B3LYP/6-31G(d) vertical ionization potentials and B3LYP/6-31G(d) HOMO LUMO gaps are largest within the "fully benzenoid" series, where all carbon atoms are members of a single sextet. The largest members of the four series studied are predicted to exhibit semiconducting properties. PMID- 12769586 TI - Stereoselective association of binuclear metallacycles in coordination polymers. AB - A series of structurally related binuclear metallacycles [Cd(NO(3))(2)L](2), where L is an angular exo-bidentate ligand, have been synthesized. Each metallacycle contains two coordinatively unsaturated, chiral metal centers within a single molecule, and the assembly of these metallacycles into polymeric framework structures has been studied systematically for the first time. Stereoselective homochiral association of [Cd(NO(3))(2)L](2) leads to the formation of helical coordination polymers, whereas meso type association results in nonhelical chain structures. The type of stereoselective aggregation depends on the conditions of self-assembly as well as on ligand functionality. Both helical and nonhelical polymeric complexes have been isolated for the metallacycle [Cd(NO(3))(2)(2,4'-pyacph)](2) (2,4'-pyacph = 2,4'-(4 ethynylphenyl)bipyridyl). Homochiral association results in the formation of helical [Cd(NO(3))]( infinity ) chains which link the binuclear [Cd(NO(3))(2)(2,4'-pyacph)](2) metallacycles into racemic two-dimensional sheets which contain both P and M [Cd(NO(3))]( infinity ) helices. In contrast, meso association leads to the formation of nonhelical one-dimensional chains. It is shown that the product of homochiral association is predominately formed at room temperature and that of meso-association is generated at elevated temperatures. Thus, it may be concluded that the homochiral association appears to be energetically less favorable than the meso-association, a conclusion that has been confirmed by theoretical calculations of the crystal lattice energy. Several high-yield syntheses of bipyridyl-type ligands used for metallacyclic assembly are also reported. PMID- 12769587 TI - Atomistic molecular dynamics simulations of chemical force microscopy. AB - Chemical force microscopy and related force measurement techniques have emerged as powerful tools for studying fundamental interactions central to understanding adhesion and tribology at the molecular scale. However, detailed interpretation of these interactions requires knowledge of chemical and physical processes occurring in the region of the tip-sample junction that experiments cannot provide, such as atomic-scale motions and distribution of forces. In an effort to address some of these open issues, atomistic molecular dynamics simulations were performed modeling a chemical force microscope stylus covered with a planar C12 alkylthiolate self-assembled monolayer (SAM) interacting with a solid wall. A complete loading-unloading sequence was simulated under conditions of near constant equilibrium, approximating the case of infinitely slow tip motion. In the absence of the solid wall, the stylus film existed in a fluid state with structural and dynamic properties similar to those of the analogous planar SAM at an elevated temperature. When the wall was brought into contact with the stylus and pressed against it, a series of reversible changes occurred culminating with solidification of the SAM film at the largest compressive force. During loading, the chemical composition of the contact changed, as much of the film's interior was exposed to the wall. At all tip heights, the distribution of forces within the contact zone was uneven and subject to large local fluctuations. Analysis using the Johnson-Kendall-Roberts, Derjaguin-Muller-Toporov, and Hertz contacts mechanics models revealed significant deviations from the simulation results, with the JKR model providing best overall agreement. Some of the discrepancies found would be overlooked in an actual experiment, where, unlike the simulations, contact area is not separately known, possibly producing a misleading or incorrect interpretation of experimental results. These shortcomings may be improved upon by using a model that correctly accounts for the finite thickness of the compliant components and nonlinear elastic effects. PMID- 12769588 TI - Symmetry and bonding in metalloporphyrins. A modern implementation for the bonding analyses of five- and six-coordinate high-spin iron(III)-porphyrin complexes through density functional calculation and NMR spectroscopy. AB - Bonding interactions between the iron and the porphyrin macrocycle of five- and six-coordinate high-spin iron(III)-porphyrin complexes are analyzed within the framework of approximate density functional theory with the use of the quantitative energy decomposition scheme in combination with removal of the vacant pi orbitals of the porphyrin from the valence space. Although the relative extent of the iron-porphyrin interactions can be evaluated qualitatively through the spin population and orbital contribution analyses, the bond strengths corresponding to different symmetry representations can be only approximated quantitatively by the orbital interaction energies. In contrast to previous suggestions, there are only limited Fe --> P pi back-bonding interactions in high spin iron(III)-porphyrin complexes. It is the symmetry-allowed bonding interaction between d(z)2 and a(2u) orbitals that is responsible for the positive pi spin densities at the meso-carbons of five-coordinate iron(III)-porphyrin complexes. Both five- and six-coordinate complexes show significant P --> Fe pi donation, which is further enhanced by the movement of the metal toward the in plane position for six-coordinate complexes. These bonding characteristics correlate very well with the NMR data reported experimentally. The extraordinary bonding interaction between d(z)2 and a(2u) orbitals in five-coordinate iron(III) porphyrin complexes offers a novel symmetry-controlled mechanism for spin transfer between the axial ligand sigma system and the porphyrin pi system and may be critical to the electron transfer pathways mediated by hemoproteins. PMID- 12769589 TI - Ion atmosphere relaxation and percolative electron transfer in Co bipyridine DNA molten salts. AB - Polypyridyl complexes of Co decorated with 350-Da polyether chains (Co(350)(2+)) form molten phases of nucleic acids when paired with DNA counterions (Co(350)DNA) or 25-mer oligonucleotides. Analysis of voltammetry and chronoamperometry of mixtures of these phases with complexes having ClO(4)(-) counterions (Co(350)(ClO(4))(2)) and no other diluent provides charge transport rates from the oxidation and reduction currents for the complexes. As the mole fraction of the Co(350)(ClO(4))(2) complex in the mixture is varied from ca. 0.25 to 1, the physical diffusion constants derived from the Co(III/II) wave increase from 1 x 10(-11) cm(2)/s to 5 x 10(-10) cm(2)/s, and apparent diffusion constants dominated by the Co(II/I) electron self-exchange increase from 1 x 10(-10) cm(2)/s to 2 x 10(-8) cm(2)/s. Pure Co(350)DNA melts, containing no Co(350)(ClO(4))(2) complex, do not exhibit recognizable voltammetric waves; DNA suppresses the Co(II/I) electron transfer reactions of Co complexes for which it is the counterion. There are therefore two microscopically distinct kinds of Co(350) complexes, those with DNA and those with ClO(4)(-) counterions, with respect to their Co(II/I) electron-transfer dynamics, leading to percolative behavior in their mixtures. The electron-transfer rates of the Co(II/I) couple are controlled by the diffusive relaxation of the ionic atmosphere around the reaction pair, and the inactivity of the bound Co complexes can be attributed to the very low mobility of the anionic phosphate groups in the DNA counterion. Substitution of sulfonated polystyrene for DNA produced similar results, suggesting that this phenomenon is general to other polymer counterions of low mobility. We conclude that the measured Co(II/I) charge transport and electron transfer rate constants reflect more the diffusive mobility of the perchlorate counterion than the intrinsic Co(II/I) electron hopping rate. PMID- 12769590 TI - Theoretical study of the magnetic behavior of hexanuclear Cu(II) and Ni(II) polysiloxanolato complexes. AB - A theoretical density functional study of the exchange coupling in hexanuclear polysiloxanolato-bridged complexes of Cu(II) and Ni(II) is presented. By calculating the energies of three different spin configurations, we can obtain estimates of the first-, second-, and third-neighbor exchange coupling constants. The study has been carried out for the complete structures of the Cu pristine cluster and of the chloroenclathrated Ni complex as well as for the hypotethical pristine Ni compound and for magnetically dinuclear analogues M(2)Zn(4) (M = Cu, Ni). PMID- 12769591 TI - Relationships between temperature, magnetic moment, and continuous symmetry measures in spin crossover complexes. AB - A study of the changes in the symmetry of the metal coordination sphere that accompany spin crossover transitions has been carried out. Changes in symmetry have been calibrated by using the continuous symmetry measures approach. Different behaviors have been found for several families of complexes, depending on the nature of their ligands. The most remarkable changes take place for trischelate complexes, for which octahedricity, trigonal prismacity, chirality, bite angle, bond distance, tempeature, pressure, magnetic moment, and density are correlated. PMID- 12769592 TI - A biofuel cell with electrochemically switchable and tunable power output. AB - An electroswitchable and tunable biofuel cell based on the biocatalyzed oxidation of glucose is described. The anode consists of a Cu(2+)-poly(acrylic acid) film on which the redox-relay pyrroloquinoline quinone (PQQ) and the flavin adenine dinucleotide (FAD) cofactor are covalently linked. Apo-glucose oxidase is reconstituted on the FAD sites to yield the glucose oxidase (GOx)-functionalized electrode. The cathode consists of a Cu(2+)-poly(acrylic acid) film that provides the functional interface for the covalent linkage of cytochrome c (Cyt c) that is further linked to cytochrome oxidase (COx). Electrochemical reduction of the Cu(2+)-poly(acrylic acid) films (applied potential -0.5 V vs SCE) associated with the anode and cathode yields the conductive Cu(0)-poly(acrylic acid) matrixes that electrically contact the GOx-electrode and the COx/Cyt c-electrode, respectively. The short-circuit current and open-circuit voltage of the biofuel cell correspond to 105 microA (current density ca. 550 microA cm(-2)) and 120 mV, respectively, and the maximum extracted power from the cell is 4.3 microW at an external loading resistance of 1 kOmega. The electrochemical oxidation of the polymer films associated with the electrodes (applied potential 0.5 V) yields the nonconductive Cu(2+)-poly(acrylic acid) films that completely block the biofuel cell operation. By the cyclic electrochemical reduction and oxidation of the polymer films associated with the anode and cathode between the Cu(0) poly(acrylic acid) and Cu(2+)-poly(acrylic acid) states, the biofuel cell performance is reversibly switched between "ON" and "OFF" states, respectively. The electrochemical reduction of the Cu(2+)-polymer film to the Cu(0)-polymer film is a slow process (ca. 1000 s) because the formation and aggregation of the Cu(0)-clusters requires the migration of Cu(2+) ions in the polymer film and their reduction at conductive sites. The slow reduction of the Cu(2+)-polymer films allows for the controlling of the content of conductive domains in the films and the tuning of the output power of the biofuel cell. The electron transfer resistances of the cathodic and anodic processes were characterized by impedance spectroscopy. Also, the overall resistances of the biofuel cell generated by the time-dependent electrochemical reduction process were followed by impedance spectroscopy and correlated with the internal resistances of the cell upon its operation. PMID- 12769593 TI - Photodetachment of zwitterions: probing intramolecular Coulomb repulsion and attraction in the gas phase using mono-decarboxylated pyridinium dicarboxylates. Implications on the mechanism of orotidine 5'-monophosphate decarboxylase. AB - Negative ion photoelectron spectra resulting from the decarboxylation of nine zwitterionic pyridinium dicarboxylates (D(x,y)) are reported. Structural assignments are made on the basis of analogy to the spectra of related species, labeling experiments with (13)C- or (2)H-containing substrates, independent syntheses, and comparison to density functional theory and ab initio (B3LYP and CCSD(T), respectively) results. In some cases, an acid-catalyzed isomerization of the D(x,y)-CO(2) ions was found to take place. Adiabatic detachment energies of the resulting zwitterionic ions were measured and are well reproduced by theory. The relative stabilities of the D(x,y)-CO(2) decarboxylation products are largely determined by their intramolecular electrostatic interactions, which are directly probed by the photoelectron spectra and were analyzed in terms of the resulting Coulombic forces. Expulsion of carbon dioxide from the D(x,y) ions was also used as an electrostatic model to probe the mechanism of the enzyme-catalyzed conversion of orotidine 5'-monophosphate (OMP) to uridine 5'-monophosphate (UMP). It was found that the loss of CO(2) from these zwitterions and from oxygen protonated OMP is retarded by the presence of an additional anionic group. This suggests that the formation of a zwitterion intermediate in the enzyme-catalyzed transformation of OMP to UMP may have less of an energetic impact than commonly thought and could be a "red herring". If so, the electrostatic stress mechanism proposed by Larsen et al. and Pai, Guo, and co-workers maybe followed. PMID- 12769594 TI - Measurement of multiple psi torsion angles in uniformly 13C,15N-labeled alpha spectrin SH3 domain using 3D 15N-13C-13C-15N MAS dipolar-chemical shift correlation spectroscopy. AB - We demonstrate the simultaneous measurement of several backbone torsion angles psi in the uniformly (13)C,(15)N-labeled alpha-Spectrin SH3 domain using two different 3D 15N-13C-13C-15N dipolar-chemical shift magic-angle spinning (MAS) NMR experiments. The first NCCN experiment utilizes double quantum (DQ) spectroscopy combined with the INADEQUATE type 13C-13C chemical shift correlation. The decay of the DQ coherences formed between 13C'(i) and 13C(alphai) spin pairs is determined by the "correlated" dipolar field due to 15N(i)-13C(alphai) and 13C'(i)-15N(i+1) dipolar couplings and is particularly sensitive to variations of the torsion angle in the regime |psi| > 140 degrees. However, the ability of this experiment to constrain multiple psi-torsion angles is limited by the resolution of the 13C(alpha)-(13)CO correlation spectrum. This problem is partially addressed in the second approach described here, which is an NCOCA NCCN experiment. In this case the resolution is enhanced by the superior spectral dispersion of the 15N resonances present in the 15N(i+1)-13C(alphai) part of the NCOCA chemical shift correlation spectrum. For the case of the 62 residue alpha-spectrin SH3 domain, we determined 13 psi angle constraints with the INADEQUATE NCCN experiment and 22 psi constraints were measured in the NCOCA NCCN experiment. PMID- 12769595 TI - Galanin: a biologically active peptide. AB - Galanin is a biologically active neuropeptide, widely distributed in the central and peripheral nervous systems and the endocrine system. The amino acid sequence of galanin is very conserved (almost 90% among species), indicating the importance of the molecule. Galanin has multiple biological effects. In the central nervous system, galanin alters the release of several neurotransmitters. In particular the ability of galanin to inhibit acetylcholine release, along with the observation of hyperinervation of galanin fibres in the human basal forebrain of Alzheimer's disease patients, suggest a possible role for galanin in the cholinergic dysfunction, characteristic of the disease. Moreover, galanin has been suggested to be involved in other neuronal functions, such as learning and memory, epileptic activity, nociception, spinal reflexes and feeding. Galanin has also been shown to increase the levels of growth hormone, prolactin and luteinizing hormone, to inhibit glucose induced insulin release and to affect gastrointestinal motility. The expression of galanin (mRNA and peptide levels) is elevated following estrogen administration, neuronal activation, denervation and/or nerve injury, as well as during development. The spectrum of galanin's activities indicates that galanin is an important messenger for intercellular communication within the nervous system and the neuroendocrine axis. Galanin acts at specific membrane receptors to exert its effects; so far three human and rodent galanin receptor subtypes have been cloned. Galanin agonists have been shown to have therapeutic application in treatment of chronic pain; galanin antagonists have therapeutic potential in treatment of Alzheimer's disease, depression, and feeding disorders. PMID- 12769596 TI - The brain endothelin system as potential target for brain-related pathologies. AB - The endothelin system, consisting of three peptides, two peptidases and two G protein coupled receptors, is widely expressed in the brain cell types and brain derived tumor cell lines. The stimulation of endothelin receptors elicits a variety of short- and long-term changes at cellular level but the effects of the modulation of the endothelin system in brain physiology and pathophysiology are, at the present time, poorly understood. Altered expression of endothelins (ETs) in reactive astrocytes has been observed in many pathological conditions of the human brain, such as infarcts, lacunae, traumatic conditions, Alzheimer's disease and inflammatory diseases of the brain. In addition, recent studies have shown that endothelin antagonists might inhibit growth and induce cell death in human melanoma cells in vitro and in vivo, and have emphasized a possible role of endothelin peptides as autocrine or paracrine factor in the proliferation and dissemination of tumor cell lines. Given the fact that brain cell and a variety of brain tumor cell lines express all the components of the endothelin system, further studies are warranted to demonstrate a possible therapeutic role of endothelin antagonist in the pharmacological treatment of brain diseases and brain tumors. PMID- 12769597 TI - Disorders of the circadian clock: etiology and possible therapeutic targets. AB - The mammalian circadian clock in the suprachiasmatic nuclei (SCN) of the hypothalamus conveys 24-hr rhythmicity to sleep-wake cycles, temperature, locomotor activity and virtually all other behavioral and physiological processes. In order for these cycles to be adaptive, they must be synchronized, or entrained, to the 24-hr light/dark cycle produced by the rotation of the Earth. The timing of circadian variables relative to the light/dark cycle, i.e., the phase angle of entrainment, is influenced by intrinsic circadian clock properties that are to an extent genetically determined, and thus varies between individuals. In extreme cases (advanced or delayed sleep phase syndrome) or during shift work or jet lag, the phase angle of entrainment may be incompatible with work requirements or other social demands, resulting in negative consequences to health and productivity. This review describes the etiology of circadian disorders within the context of formal circadian clock properties and summarizes studies in humans and in other species which link specific genetic loci to circadian clock function and malfunction. The proteins encoded by these genetic loci play key roles in the intracellular feedback loop that generates circadian rhythms, and thus represent therapeutic targets for the treatment of both endogenous and exogenous circadian disorders. PMID- 12769598 TI - The plasma membrane: a target and hurdle for the development of anti-Abeta drugs? AB - The plasma membrane has been the subject of intense investigation in the search for anti-amyloidogenic drugs for the treatment of Alzheimer's disease. Studies have highlighted numerous toxic properties of the well-known amyloid Abeta peptide on neuronal membranes. In this respect recent experimental data suggest that an early step in amyloid toxicity might be intracellularly mediated. This suggests that effective anti-amyloidogenic agents must be able to readily cross the plasma membrane while at the same time, counteracting the deleterious effects of the Abeta peptide on the phospholipid bilayer. This review summarizes recent findings regarding amyloid-plasma membrane interactions and discusses their relevance for the design of novel, potential anti-Abeta drugs. PMID- 12769599 TI - Depressed or demented: common CNS drug targets?! AB - A body of evidence emerging in antidepressant and antidementia research has revealed a convergence of molecular events known to regulate neuronal plasticity in learning and memory with molecular actions of drugs for the treatment of depression. Many antidepressants are reported to have positive impact on learning and memory. These include agents acting through monoaminergic neurotransmitter systems, non-monoaminergic transmitter systems, and hormones. On the other hand, agents that appear to have memory-enhancing or antidementia value are frequently found to exhibit antidepressant activity in patients and animal depression models. It is becoming clear that the comorbidity of depression and dementia does not occur by chance, but rather is an inevitable consequence of pathologic relationships between the conditions. Molecular mechanisms and cascades that underlie memory may be shared by mood regulation and are vulnerable to stress and injuries. This review focuses on recent findings regarding effects of a variety of agents on dementia and depression and their common molecular mechanisms as well as their differences. A better understanding of the key underlying molecular components whose changed activities have dramatic influences on mood and cognition may lead to the development of novel and more effective therapeutic agents for the treatment of depression and dementia. In this review, some of the recent findings that suggest novel therapeutic strategies are also highlighted. PMID- 12769600 TI - Organophosphate induced delayed polyneuropathy. AB - This review discusses the current understanding of organophosphate induced delayed polyneuropathy (OPIDP) with emphasis on molecular mechanisms, pathogenesis and possibilities for prevention/therapy. OPIDP is a rare toxicity caused by certain organophosphorus compounds (OP) characterized by degeneration of some long axons in the central and peripheral nervous system that appear about 2-3 weeks after exposure. The molecular target for OPIDP is considered to be an enzyme in the nervous system known as neuropathy target esterase (NTE). NTE can be inhibited by two types of inhibitors: a) phosphates, phosphonates, and phosphoramidates, which cause OPIDP when >70% of the enzyme is inhibited, and b) phosphinates, carbamates, and sulfonyl halides which inhibit NTE and cause either protection from, or promotion, of OPIDP when given before or after a neuropathic OP, respectively. The ability of a NTE inhibitor to cause OPIDP, besides its affinity for the enzyme, is related to its chemical structure and the residue left attached to the NTE. If such residues undergo the aging reaction i.e. the loss of an alkyl group bound to the enzyme, those OPs usually have a high likelihood of causing OPIDP. Protection from neuropathic doses of OP inhibitors is obtained when NTE is inhibited with nonageable inhibitors. Promotion of OPIDP involves another site besides NTE because it can occur when all NTE is affected. It is now known that this other site is similar to NTE in that it is also sensitive to mipafox but at much higher concentrations. Promotion affects either the progression or expression of OPIDP after the initial biochemical effect on NTE. Some recent observations suggest that development of OPIDP in hens can be influenced by atropine, oximes and methylprednisolone when they are given before or soon after neuropathic OPs. PMID- 12769601 TI - Non-peptidic CRF1 receptor antagonists for the treatment of anxiety, depression and stress disorders. AB - Anxiety and depression are psychiatric disorders that constitute a major health concern worldwide, and new pharmacological approaches with the potential for improved efficacy and decreased side effect profiles relative to currently marketed drugs are desired. Since the identification of corticotropin releasing factor (CRF) by Vale and colleagues in 1981, an extensive research effort has solidified the importance of this 41 amino acid peptide in mediating the body's behavioral, endocrine, and autonomic responses to stress. The further identification of CRF receptor subtypes has provided compelling targets for novel pharmaceutical agents. The present review focuses on the potential of non peptidic antagonists of the CRF(1) receptor subtype as a novel therapeutic approach for the treatment of anxiety and depression. The first section reviews preclinical and clinical evidence implicating CRF, in general, and CRF(1) receptors, in particular, in anxiety and depression. Clinical studies have demonstrated a dysfunctional hypothalamic-pituitary-adrenal (HPA) axis and/or elevated CRF levels in depression and in some anxiety disorders. Preclinical data utlilizing correlational methods, genetic models, and exogenous CRF administration techniques in rodents and non-human primates supports a link between hyperactive CRF pathways and anxiogenic and depressive-like symptoms. Studies employing the use of receptor knockouts and selective, non-peptidic antagonists of the CRF(1) receptor have demonstrated anxiolytic and antidepressant effects under certain types of laboratory conditions. A Phase II, open-label, clinical trial in major depressive disorder has reported that a CRF(1) receptor antagonist was safe and effective in reducing symptoms of anxiety and depression. In the second section, a topological approach is used to describe the design strategies employed to produce potent, non-peptidic CRF(1) receptor antagonists. Two main topologies, featuring a center core, a top side-chain, and a pending aromatic ring, can be used to characterize the vast majority of currently known CRF(1) receptor antagonists. By exploiting some of these structural elements, pharmacological, physicochemical, and pharmacokinetic properties can be modulated and optimized. However, as a result of a relatively conservative iteration process during the structural optimization, the chemical space presently defined by the existing CRF(1) receptor antagonists still remains fairly narrow. Expanding these structural and topological boundaries, while optimizing the "drug-like" properties of the CRF(1) receptor antagonists, seems to be a common objective across pharmaceutical companies to maximize the chances for a clinical success in the near future. PMID- 12769602 TI - Cholecystokinin-8 and nerve growth factor: two endogenous molecules working for the upkeep and repair of the nervous system. AB - Cholecystokinin-8 (CCK-8), the small peptide initially described as a gastric factor involved in the regulation of feeding behavior, is today recognized as one of the most abundant neurotransmitters/ neuropeptides in brain and is an important signal factor for the peripheral and central nervous systems. In the past twenty years, many studies have focused on possible clinical applications of this peptide and its receptor ligands in psychiatric diseases and gastrointestinal pathologies. Recently it has been suggested that CCK-8 may also have a neuroprotective role, thus opening a new field of interest around the physiology and the pharmacology of this neuropeptide and its receptors. It has been demonstrated that CCK-8 counteracts neuronal deficit following chemical or surgical lesions in both the central and peripheral nervous systems and that Nerve Growth factor (NGF) is involved in the CCK-induced recovery process. By using selective CCK receptor antagonists it has been demonstrated that CCK-8, when injected intraperitoneally, has the ability to stimulate NGF synthesis in brain and peripheral organs by a mechanism that involves the activation of CCK receptors. As has been widely reported, NGF is an essential survival and differentiative factor for selective neuronal populations of the PNS and CNS and plays a role in the events of degeneration and repair of the nervous system in diseases with different etiologies, e.g. neurodegenerative and autoimmune diseases as well as diabetes-associated pathologies. The possibility of using NGF in therapy has been evaluated and systemic and intracerebral NGF treatment have been tested in patients and animal models. Although the results of these studies are encouraging, the difficulty to predict and/or eliminate the side effects of NGF/NGF antibody treatment has made it difficult to fully evaluate the potential of the beneficial effects. In this context recent results obtained in our laboratories may offer a new prospective for the pharmacological approaches to the diseases associated with altered NGF production and functions. The data of our recent observations on NGF and CCK-8 is covered in this review. PMID- 12769603 TI - Molecular and cellular mechanism of glutamate receptors in relation to amyotrophic lateral sclerosis. AB - Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder of the central nervous system (CNS) with an unknown etiology. This disorder is characterized clinically by muscular weakness and wasting, and pathologically by selective degeneration of the corticospinal tracts and motor neurons of the brain stem and spinal cord. Median survival following onset is 3 to 5 years. Riluzole, an antiglutamatergic agent has been shown to have modest beneficial effects on survival. Glutamate is the main excitatory neurotransmitter in the CNS and excessive activation of glutamate receptors is excitotoxic to neurons. Glutamate receptor-mediated excitotoxicity has been proposed to explain the pattern of selective neuronal cell death and clinical manifestation of ALS. Activation of glutamate receptors leading to elevation of intracellular calcium may play a major role. This review will focus on the current understanding of the molecular and cellular mechanisms of glutamate receptors in relation to ALS. PMID- 12769604 TI - Serotonin transporters. AB - The serotonin transporter is most well known as the site of action of the serotonin reuptake inhibitors, which were initially developed as antidepressants, but now are the most widely used agents in the treatment of many additional neuropsychiatric and related disorders. The discovery that the serotonin transporter gene possesses a functional polymorphism within the promoter region led to many studies examining this polymorphism in individuals with different neuropsychiatric disorders. These observations raise the possibility of developing a new class of agents with different therapeutic and side effect profiles than conventional drugs. This overview of recent studies of the serotonin transporters attempts to open new areas for discussion and define concepts not yet comprehensively reviewed. PMID- 12769605 TI - Mutational analysis of the serotonergic system: recent findings using knockout mice. AB - Serotonin modulates numerous processes in the central nervous system related to anxiety and fear, mood, aggression, sleep, ingestive behaviors, reward systems, and psychosis. Serotonergic dysfunction has been implicated in several neuropsychiatric conditions but efforts to develop more specific pharmacological agents have been hampered by the complexity of this system at the receptor level. There are at least 14 distinct receptors that mediate the effects of serotonin as well as several enzymes that control its synthesis and metabolism but very few pharmacological agents are able to selectively target a single receptor. Several groups including ours have used a genetic strategy to ablate specific serotonin receptors in an effort to dissect out their functions in the central nervous system. The strength of this approach is the high degree of specificity that can be achieved in the knockout (gene inactivation) approach since a single receptor gene can be selectively targeted. To date several inactivation mutations of specific serotonin receptors have been generated producing interesting behavioral phenotypes related to anxiety, depression, drug abuse, psychosis, and cognition. In many cases knockout mice have been able to confirm what has already been suspected based on pharmacological studies. In other instances, mutations have demonstrated new functions of serotonergic genes in development and behavior. In this review, the current literature regarding phenotypic changes in mice bearing inactivation mutations of serotonin receptors, the serotonin transporter, and the monoamine oxidase A will be discussed and major findings emphasized. PMID- 12769606 TI - Knock-out and knock-in mice to investigate the role of nicotinic receptors in the central nervous system. AB - Pharmacological manipulations of nicotinic transmission have long been the only way to investigate the role of nicotinic acetylcholine receptors (nAChRs) in the brain. More recently, however, the use of genetically engineered knock-out (Ko) and knock-in (Kin) mice has provided a powerful alternative to the classical pharmacological approach. These animal models are not only useful in order to re examine and refine the results derived from pharmacological studies, but they also provide a unique opportunity to determine the subunit composition of native receptors involved in various aspects of nicotinic transmission. Ultimately, this knowledge will be extremely valuable in the process of designing new drugs that can mimic the beneficial effects of nicotine for the treatment of certain neuropathologies or that may be useful in smoking cessation therapies. In this review, we present recent data obtained from studies of mutant mice that have contributed to our understanding of the role and composition of nAChRs in the central nervous system (CNS). The advantages and pitfalls of Ko models will also be discussed. PMID- 12769607 TI - Nicotinic receptor mutant mice in the study of autonomic function. AB - Nicotinic acetylcholine receptors (nAChRs) have been implicated in Alzheimer's disease, Parkinson's disease, epilepsy, nicotine addiction, schizophrenia, and autonomic dysfunctions. Although nicotine may be used therapeutically either alone or in association with other drugs, its beneficial effects are limited by its addictive properties and a number of other side effects. A deeper understanding of nicotinic cholinergic mechanisms is necessary to develop nAChR ligands that are more selective, less toxic, and more therapeutically effective than nicotine. Although there has been significant progress identifying the nAChR subunits that form functional nAChRs, there is limited information associating the location and function of nAChR subtypes in the nervous system. Several groups have genetically engineered mice in which one or more genes encoding nAChR subunits has been deleted or altered. Mice with nAChR mutations targeted to subunits that are highly expressed in the CNS have brought insight into the nAChR mechanisms involved in nicotine addiction, analgesia, aging, and nicotine-induced behaviors. Mutations targeted to nAChR subunits that are highly expressed in the peripheral nervous system have opened a window on the complex mechanisms governing autonomic control of peripheral organs. This review examines nAChRs in the autonomic control of peripheral organ systems as gleaned from studies of nAChR mutant mice. PMID- 12769608 TI - Recent progress in the development of subtype selective nicotinic acetylcholine receptor ligands. AB - Nicotinic acetylcholine receptors (nAChRs) are pentameric ligand gated ion channels, which are found at the neuromuscular junction and in the central and peripheral nervous systems. The channels can be assembled from fourteen known subunits. The exact combination and function of all the channels are still not determined but in the CNS certain combinations have been identified which appear to modulate the release of specific neurotransmitters. Non-specific nAChR agonists like nicotine and epibatidine, have been shown to have interesting pharmacology but their clinical value is limited by their undesirable side effects. Selective ligands for different receptor subtypes have been reported and these compounds are probably the best tools for determining the function of the subtypes. The expectation is that some receptor subtype selective nAChR ligands will be clinically useful for the treatment of a broad range of CNS disorders. The development of stable cell lines functionally expressing specific combinations of subunits has greatly improved our understanding of ligand specificity. There have also been advances in the modelling of the ligand binding site, thanks to the discovery of a homologous snail ACh binding protein the X-ray structure of which was determined in 2001. These techniques should lead to rapid advances in the development of truly subtype selective ligands. In this review we describe recent progress in the area and describe the first 1000 fold selective low molecular weight ligands from the AstraZeneca group. We also comment on the first subtype specific channel modulators. PMID- 12769610 TI - Regulation of nicotinic acetylcholine receptor numbers and function by chronic nicotine exposure. AB - Recent advances concerning effects of chronic nicotine exposure on nicotinic acetylcholine receptor (nAChR) expression are reviewed. Implications are assessed of these findings for roles of nAChR in health and disease and for design of drugs for treatment of neurological and psychiatric disorders. Most studies continue to show that chronic nicotine exposure induces increases in numbers of nAChR-like binding or antigenic sites ("upregulation") across all nAChR subtypes investigated, but with time- and dose-dependencies and magnitudes for these effects that are unique to subsets of nAChR subtypes. These effects appear to be post-transcriptionally based, but mechanisms involved remain obscure. With notable exceptions, most studies also show that chronic nicotine exposure induces several phases of nAChR functional loss ("desensitization" and longer-lasting "persistent inactivation") assessed in response to acute nicotinic agonist challenges. Times for onset and recovery and dose-dependencies for nicotine induced functional loss also are nAChR subtype-specific. Some findings suggest that upregulation and functional loss are not causally- or mechanistically related. It is suggested that upregulation is not as physiologically significant in vivo as functional effects of chronic nicotine exposure. By contrast, brain levels of nicotine in tobacco users, and perhaps levels of acetylcholine in the extracellular space, clearly are in the range that would alter the balance between nAChR in functionally ready or inactivated states. Further work is warranted to illuminate how effects of chronic nicotinic ligand exposure are integrated across nAChR subtypes and the neuronal circuits and chemical signaling pathways that they service to produce nicotine dependence and/or therapeutic benefit. PMID- 12769609 TI - Targeting neuronal nicotinic receptors: a path to new therapies. AB - The structural heterogeneity, the ubiquity of anatomical distribution, and the demonstrated modulation of biological functions is consistent with the view that nicotinic cholinergic signaling plays a key regulatory role in the brain and influences a number of neuronal processes including sensory processing, motor activity, and cognitive function. It has become evident that perturbation of nicotinic cholinergic neurotransmission can result in diverse CNS pathologies providing the potential for therapeutic intervention in a number of neurodegenerative, neuropsychiatric and neurological disorders. This review will provide a status on the rationale for neuronal therapies targeting neuronal nicotinic receptors (nAChRs) and discusses the multifaceted beneficial effects than can be achieved through manipulation of cholinergic pharmacology. Recent advances and issues relating to rational drug design based on the structure of acetylcholine binding protein are discussed. PMID- 12769611 TI - Human brain nicotinic receptors, their distribution and participation in neuropsychiatric disorders. AB - Mapping of nicotinic acetylcholine receptor (nAChR) subtypes and subunits in human brain is far from complete, however it is clear that multiple subunits are present (including alpha3, alpha4, alpha5, alpha6 and alpha7, beta2, alpha3 and beta4) and that these receptors are not solely distributed on neurones, but also on cerebral vasculature and astrocytes. It is important to elucidate subunit composition of receptors associated with different cell types and pathways within the human CNS in terms of potential nicotinic therapy for a range of both developmental and age-related disorders in which nAChR attenuation occurs. Reductions in nAChRs are reported in Alzheimer's and Parkinson's diseases, dementia with Lewy bodies, schizophrenia and autism, but may not be associated with reduced cortical cholinergic innervation observed in vascular dementia or occur at an early stage in Down's syndrome. Changes in nAChR expression in neuropsychiatric disorders appear to be brain region and subtype specific and have been shown in some instances to be associated with pathology and symptomatology. It is likely that deficits in alpha4-containing receptors predominate in cortical areas in Alzheimer's disease and autism, whereas reduction of alpha7 receptors may be more important in schizophrenia. Changes in astrocytic and vascular nAChR expression in neurodegenerative diseases should also be considered. Studies using both animal models and human autopsy tissue suggest that nAChRs can play a role in neuroprotection against age-related pathology. It is possible that the development of nAChR subtype specific drugs may lead to advances in therapy for both age-related and psychiatric disorders. PMID- 12769612 TI - The role of neuronal nicotinic acetylcholine receptors in acute and chronic neurodegeneration. AB - In the last five years there has been a rapid explosion of publications reporting that neuronal nicotinic acetylcholine receptors (nAChRs) play a role in neurodegenerative disorders. Furthermore, there is a well-established loss of nAChRs in post-mortem brains from patients with Alzheimer's disease, Parkinson's disease and a range of other disorders. In the present review we discuss the evidence that nicotine and subtype selective nAChR ligands can provide neuroprotection in in vitro cell culture systems and in in vivo studies in animal models of such disorders. Whilst in vitro data pertaining to a protective effect of nicotine against nigral neurotoxins like MPTP is less robust, most studies agree that nicotine is protective against glutamate and beta-amyloid toxicity in various culture systems. This effect appears to be mediated by alpha7 subtype nAChRs since the protection is blocked by alpha-bungarotoxin and is mimicked by alpha7 selective agonists. In vivo studies indicate that alpha7 receptors play a critical role in protection from cholinergic lesions and enhancing cognitive function. The exact subtype involved in the neuroprotectant effects seen in animal models of Parkinson's disease is not clear, but in general broad spectrum nAChR agonists appear to provide protection, while alpha4beta2 receptors appear to mediate symptomatic improvements. Evidence favouring a protectant effect of nicotine against acute degenerative conditions is less strong, though some protection has been observed with nicotine pre-treatment in global ischaemia models. A variety of cellular mechanisms ranging from the production of growth factors through to inactivation of toxins and antioxidant actions of nicotine have been proposed to underlie the nAChR-mediated neuroprotection in vitro and in vivo. In summary, although the lack of subtype selective ligands has hampered progress, it is clear that in the future neuronal nAChR agonists could provide functional improvements and slow or halt the progress of several crippling degenerative diseases. PMID- 12769613 TI - Neuroplasticity within the mesoaccumbens dopamine system and its role in tobacco dependence. AB - The development of nicotine dependence is related to stimulation of the dopamine projections to the nucleus accumbens. This review considers the evidence that the addictive potential of nicotine depends upon its ability to elicit burst firing of these neurones and, thereby, evoke a large and sustained increase in the dopamine concentration in the extracellular space between the cells. This dopamine, it is argued, stimulates extra-synaptic dopamine receptors that mediate the responses underling the development of dependence. The review also considers the hypothesis that the two principal subdivisions of the structure, the core and shell, play different roles in the development of dependence. It proposes that the projections to the shell signal the presence of a rewarding stimulus and facilitate the acquisition of behaviours related to obtaining the reward. In contrast, the projections to the core, which are sensitised selectively by repeated exposure to the drug, mediate the transition to habit or Pavlovian responding to cues repetitively paired with the positive reinforcing properties of nicotine. Nicotine withdrawal, following a period of chronic exposure, diminishes the activity of the dopamine projections to the accumbal shell, a response that is thought to be the neural correlate of the anhedonia experienced by many abstinent smokers. The data suggest that plasticity within the principal mesoaccumbens dopamine projections play a central role in the development of nicotine dependence and that the mechanisms underlying the plasticity may provide putative targets for the treatment of tobacco dependence. PMID- 12769614 TI - Nicotinic treatment for cognitive dysfunction. AB - Nicotinic medications may provide beneficial therapeutic treatment for cognitive dysfunction such as Alzheimer's disease, schizophrenia and attention deficit hyperactivity disorder (ADHD). For development of nicotinic treatments we are fortunate to have a well characterized lead compound, nicotine. Transdermal nicotine patches offer a way to deliver measured doses of nicotine in a considerably safer fashion than the more traditional means of administration, tobacco smoking. We have found that transdermal nicotine significantly improves attentional function in people with Alzheimer's disease, schizophrenia or ADHD as well as normal nonsmoking adults. To follow-up on this proof of principal that nicotinic treatment of cognitive dysfunction holds promise, it is important to use animal models to determine the critical neurobehavioral bases for nicotinic involvement in cognitive function so that more selective nicotinic analogues that improve cognitive function with fewer side effects can be developed. We have found with local infusion in rat studies that the hippocampus and amygdala are important substrates for nicotinic effects on working memory function. Both alpha7 and alpha4beta2 nicotinic receptors are involved in working memory. Nicotinic interactions with dopaminergic and glutaminergic systems are also important in the basis of cognitive function. Studies of the neural nicotinic mechanisms underlying cognitive function are key for opening avenues for development of safe and effective nicotinic treatments for cognitive dysfunction. PMID- 12769615 TI - The importance of nicotinic acetylcholine receptors in schizophrenia, bipolar disorder and Tourette's syndrome. AB - As the prevalence of tobacco use has decreased, it has become clear that individuals with mental illness comprise a substantial portion of the remaining smokers. Seventy to eighty percent of patients with schizophrenia smoke and their smoking is established before their first psychotic episodes or the initiation of treatment. Many patients with schizophrenia, and approximately 50% of their first degree relatives have abnormalities in auditory sensory gating and/or smooth pursuit eye movements. These abnormalities are corrected by nicotine, and they appear to be transmitted as autosomal dominant traits. Evidence is accumulating that these abnormalities reflect genetic variations in nicotine receptor number and function, that may increase susceptibility for schizophrenia. Recent studies suggest that bupropion, added to treatment with an atypical antipsychotic, can enhance the likelihood of smoking cessation or reduction in patients with schizophrenia. The prevalence of smoking is also substantially increased among patients with bipolar disorder, perhaps especially so among those with psychotic features. Nicotine delivered by gum or transdermal patch can provide short term relief for exacerbations of Tourette's Syndrome, but its use is limited by frequent toxicity, primarily nausea. PMID- 12769616 TI - Nicotinic acetylcholine receptors and epilepsy. AB - The neuronal nicotinic acetylcholine receptors (nAChRs) have multiple roles in the brain: they are involved in signal transduction by fast synaptic transmission, axo-axonic transmission, and in the modulation of presynaptic transmitter release. Presynaptic nAChRs can increase the release of excitatory as well as of inhibitory transmitters, and can thereby control neuronal excitability. Furthermore, nAChRs which are expressed in fetal brain might also be involved in brain morphogenesis. Thus, the genes coding for the different nAChR subunits are likely candidates for several neurological disorders. The CHRNA4- or CHRNB2 subunits of the nAChR are associated with autosomal dominant nocturnal frontal lobe epilepsy (ADNFLE), a rare monogenic type of idiopathic epilepsy. Electrophysiological studies demonstrated that ADNFLE mutations are causing both a loss-of-function and a gain-of-function in alpha4/beta2 heteropentameric nAChRs. PMID- 12769617 TI - Modulation of the basal ganglia by metabotropic glutamate receptors: potential for novel therapeutics. AB - The basal ganglia are implicated in a number of disorders including neurodegenerative motor diseases such as Huntington's and Parkinson's disease, as well as psychiatric disorders such as schizophrenia and obsessive compulsive disorder. In recent years, a great deal of effort has been focused on determining the basal ganglia circuitry that underlies normal behavior, as well as many of these syndromes. This has led to a detailed understanding of both the normal and pathophysiological flow of information through the basal ganglia, and has provided the opportunity to begin developing novel pharmacological methods of intervention by targeting neuromodulatory receptors with in the basal ganglia circuit. One group of receptors that holds much promise for several basal ganglia disorders is the metabotropic glutamate receptors. Data from behavioral, neurochemical, neuroanatomical and electrophysiological studies has begun to reveal the functional roles that the metabotropic glutamate receptors play in modulating the basal ganglia circuit, and suggests that compounds selectively targeting these receptors may provide novel therapies for a variety of disorders including Parkinson's disease, addiction, and epilepsy. PMID- 12769618 TI - The roles of metabotropic glutamate receptors in seizures and epilepsy. AB - Epilepsy is a disorder that afflicts more than 50 million people worldwide. Current antiepileptic drugs (AEDs), although effective in controlling seizures for the majority of individuals, remain far from ideal as therapeutics. There is a need for new drugs that act at different molecular targets than currently available AEDs and for new therapies designed to block the process of epileptogenesis. Because of their central role in modulating numerous physiological processes in the central nervous system, metabotropic glutamate receptors (mGluRs) have been implicated in the pathophysiology of a variety of neurological conditions including epilepsy. mGluRs represent attractive new targets for therapeutic control of seizures and interruption of the epileptogenic process. We review the involvement of mGluRs in the induction and expression of epileptic seizures, their potential roles in the process of epileptogenesis, and their altered expression and function in the epileptic human brain. PMID- 12769619 TI - Metabotropic glutamate receptors: potential drug targets for the treatment of schizophrenia. AB - Schizophrenia is a debilitating chronic psychiatric illness affecting 1% of the population. The cardinal features of schizophrenia are positive symptoms (thought disorder, hallucinations, catatonic behavior), negative symptoms (social withdrawal, anhedonia, apathy) and cognitive impairment. Although progress in elucidating the aetiology of schizophrenia has been slow, new insights on the neurochemical and neurophysiological mechanisms underlying the pathophysiology of this illness are beginning to emerge. The glutamate/N-methyl-D-aspartate (NMDA) hypofunction hypothesis of schizophrenia is supported by observations that administration of NMDA glutamate receptor antagonists such as phencyclidine (PCP) or ketamine induces psychosis in humans; moreover, decreased levels of glutamate and changes in several markers of glutamatergic function occur in schizophrenic brain. Administration of PCP or ketamine to rodents elicits an increase in locomotion and stereotypy accompanied by an increase in glutamate efflux in several brain regions. Systemic administration of group II metabotropic glutamate (mGlu) receptor agonists suppresses PCP-induced behavioral effects and the increase in glutamate efflux. Activation of group II mGlu receptors (mGlu2 and mGlu3) decreases glutamate release from presynaptic nerve terminals, suggesting that group II mGlu receptor agonists may be beneficial in the treatment of schizophrenia. In addition, pharmacological manipulations that enhance NMDA function may be efficacious antipsychotics. Selective activation of mGlu5 receptors significantly potentiates NMDA-induced responses, supporting this novel approach for the treatment of schizophrenia. The glutamate hypothesis of schizophrenia predicts that agents that restore the balance in glutamatergic neurotransmission will ameliorate the symptomatology associated with this illness. Development of potent, efficacious, systemically active drugs will help to address the antipsychotic potential of these novel therapeutics. This review will discuss recent progress in elucidating the pharmacology and function of group II mGlu and mGlu5 receptors in the context of current hypotheses on the pathophysiology of schizophrenia and the need for new and better antipsychotics. PMID- 12769620 TI - Metabotropic glutamate receptor involvement in models of acute and persistent pain: prospects for the development of novel analgesics. AB - The excitatory amino acid glutamate plays a major role in nociceptive processing. Ionotropic and metabotropic glutamate receptors are expressed in relevant areas of the brain, spinal cord and periphery that are involved in pain sensation and transmission. Activation of mGlu receptors along the pain neuraxis can result in either pronociceptive or antinociceptive behaviors depending on the subtype of mGluR and its location. The data published to date most strongly support the idea that mGlu1 antagonists might act as broad-spectrum analgesics. Several studies pointing to a functional upregulation of mGlu2/3 in chronic pain models suggest that agonists of these receptors might also be effective analgesics in certain conditions, most notably inflammation-induced hyperalgesia and allodynia. The expression of mGluRs throughout the pain neuraxis and the differing roles of the mGluRs in each of these regions makes it difficult to predict the efficacy of mGluR ligands based on in vitro or local administration studies. Potent, systemically active compounds that show mGluR subtype selectivity will be critical to undertake more detailed analyses in animal models of pain. PMID- 12769621 TI - The metabotropic glutamate receptors: structure, activation mechanism and pharmacology. AB - The metabotropic glutamate receptors are G-protein coupled receptors (GPCR) involved in the regulation of many synapses, including most glutamatergic fast excitatory synapses. Eight subtypes have been identified that can be classified into three groups. The molecular characterization of these receptors revealed proteins much more complex than any other GPCRs. They are composed of a Venus Flytrap (VFT) module where glutamate binds, connected to a heptahelical domain responsible for G-protein coupling. Recent data including the structure of the VFT module determined with and without glutamate, indicate that these receptors function as dimers. Moreover a number of intracellular proteins can regulate their targeting and transduction mechanism. Such structural features of mGlu receptors offer multiple possibilities for synthetic compounds to modulate their activity. In addition to agonists and competitive antagonists acting at the glutamate binding site, a number of non-competitive antagonists with inverse agonist activity, and positive allosteric modulators have been discovered. These later compounds share specific properties that make them good candidates for therapeutic applications. First, their non-amino acid structure makes them pass more easily the blood brain barrier. Second, they are much more selective than any other compound identified so far, being the first subtype selective molecules. Third, for the negative modulators, their non competitive mechanism of action makes them relatively unaffected by high concentrations of glutamate that may be present in disease states (e.g. stroke, epilepsy, neuropathic pain, etc.). Fourth, like the benzodiazepines acting at the GABA(A) receptors, the positive modulators offer a new way to increase the activity of these receptors in vivo, with a low risk of inducing their desensitization. The present review article focuses on the specific structural features of these receptors and highlights the various possibilities these offer for drug development. PMID- 12769622 TI - Treatment of schizophrenia and comorbid substance use disorder. AB - Comorbid alcohol and substance use disorders occur commonly among patients with schizophrenia and contribute to the morbidity of schizophrenia. These comorbid disorders add greatly to the financial costs and emotional toll that schizophrenia places on patients, families and the entire mental health system. While the basis for the increased abuse of alcohol and other substances in patients with schizophrenia have been linked by some investigators to "self medication" of negative symptoms of schizophrenia or extrapyramidal system effects of typical antipsychotics, we have presented a neurobiologic formulation suggesting that alcohol or other substances may transiently correct a dysfunction of the dopamine-mediated mesocorticolimbic pathways in patients with schizophrenia - pathways linked to brain reward circuits. This formulation further suggests that alcohol or other substances serve to transiently enhance the functioning of this circuit by improving the "signal detection" capability of the dopamine-rich mesocorticolimbic pathways. Treatment of comorbid substance use disorder in patients with schizophrenia involves careful use of psychosocial approaches aimed at fostering program participation and at enhancing the likelihood of abstinence. While the typical antipsychotics do not limit the comorbid substance use, and may actually worsen it, preliminary data suggest the novel antipsychotic clozapine may have the unusual ability to dramatically decrease alcohol and other substance use in patients with schizophrenia. It is not clear whether other novel antipsychotics share this ability of clozapine to limit alcohol and substance abuse. We have proposed that the effect of clozapine in this population may relate to its broad pharmacological effects, including its relatively weak blockade of the dopamine D2 receptor and its potent blockade of the serotonergic 5-HT2 receptor and the noradrenergic alpha 1 and alpha 2 receptors. Studies of other agents, employed in the pharmacotherapy of alcohol and substance use disorders without schizophrenia, are currently underway in patients with schizophrenia and comorbid disorders. PMID- 12769623 TI - Partial dopamine agonists and dopaminergic stabilizers, in the treatment of psychosis. AB - The early demonstration of chlorpromazine efficacy in schizophrenia and its subsequent identification as a dopamine receptor antagonist, established the only known mechanism for antipsychotic development to date. By extension, it is easy to hypothesize that any mechanism shown to reduce dopamine-mediated transmission in brain will have antipsychotic properties. The evaluation of partial dopamine agonists for antipsychotic efficacy and their application in the treatment of psychosis has derived from this background. Partial dopamine agonists at the D2 dopamine receptor, have high affinity for that receptor, but reduced intrinsic activity. These agonists have higher affinity for the presynaptic autoreceptor than for the postsynaptic receptor. Hence, these compounds reduce dopamine synthesis and release through an agonist action at the dopamine autoreceptor. Moreover, the agonists have lower intrinsic activity at the postsynaptic receptor than its natural ligand dopamine. Therefore, they diminish the dopaminergic signal at postsynaptic sites as well through delivering a reduced message; this component of drug action becomes more prominent the lower the intrinsic activity of the drug. Several partial dopamine agonists have been evaluated in schizophrenia. One of them, aripiprazole, is nearing approval for marketing. With partial dopamine agonist treatment, advantages should accrue to schizophrenia treatment in the areas of affect control and cognitive performance. PMID- 12769624 TI - Nicotinic agonists and psychosis. AB - Schizophrenia patients have insufficient inhibitory processing of identical paired auditory stimuli. This deficient "auditory gating" is thought to have physiological relevance, and its severity correlates with certain measures of both positive and negative symptoms. Schizophrenia patients also represent the heaviest smoking population subgroup. Because smoking temporarily normalizes their auditory gating deficit, this may represent a form of self-medication. Although this deficit is unresponsive to treatment with typical antipsychotic drugs, it does respond to the atypical antipsychotic clozapine. The normalization of this deficit by smoking may account for some of the intense drive to smoke that is experienced by schizophrenia patients. However, the normalizing effect of nicotine is transient and is not observed with repeated administration. Auditory gating is modulated by the alpha7 nicotinic receptor subtype, a rapidly desensitizing low-affinity nicotinic receptor. Agents that selectively activate the alpha7 receptor represent a novel class of therapeutic agents for use in the treatment of schizophrenia. Whether selective alpha7 agonists will have beneficial effects on symptoms other than the auditory gating deficit has not yet been established. The first developed alpha7-selective agonist, 3-2,4 dimethoxybenzylidene anabaseine (DMXB-A), normalizes auditory gating in three distinct animal models of the deficit. DMXB-A is a prototype for this potential new drug class, but proof-of-concept for this type of pharmacotherapy will not be available until the completion of planned clinical trials assessing DMXB-A's effects in schizophrenia patients. Additional avenues to the potential normalization of auditory gating deficits are also discussed, focusing on the novel efficacy of clozapine and the potential utility of allosteric modulators of nicotinic receptors. PMID- 12769625 TI - Muscarinic receptors as a target for drugs treating schizophrenia. AB - The family of 5 muscarinic acetylcholine receptors belongs to the superfamily of G protein coupled neurotransmitter receptors that serve in part as regulators of synaptic function. Muscarinic receptors are anatomically positioned in cortical and subcortical areas and modulate dopaminergic and glutamatergic neurotransmission thought to be dysfunctional in schizophrenia. Neurochemical studies have shown that dopamine and muscarinic receptors reciprocally modulate one another. For example, the muscarinic agonist xanomeline increases extracellular levels of dopamine and Fos expression in cortical areas greater than subcortical areas, similar to effects of atypical antipsychotics. In electrophysiological studies, xanomeline with acute and chronic administration decreased firing of the mesocorticolimbic dopamine A10 tract, but not the motoric dopamine A9 tract. Behavioral investigations have shown that muscarinic agonists, like dopamine antagonists, inhibit conditioned-avoidance responding and dopamine agonist-induced behaviors including hyperactivity, climbing behavior and disruption of prepulse inhibition, models for positive symptoms of schizophrenia. Transgenic knockout mice lacking M(4) receptors are hyperactive and hyper responsive to dopamine D(1) agonists, suggesting a dynamic balance between the dopamine and M(4) receptors. Muscarinic agonists had activity in animal models of negative symptoms, cognitive dysfunction and affective disorders, symptoms that are prominent in schizophrenic patients. Consistent with effects in animal models, preliminary clinical investigation indicates that muscarinic agonists like xanomeline may be effective in the pharmacotherapy of schizophrenia. Thus, we hypothesize that a combined M(1) agonist to promote cognition and a M(4) agonist for antipsychotic-like effects would treat the symptom domains of schizophrenia without parasympathomimetic side effects. PMID- 12769626 TI - Ionotropic glutamate receptors as therapeutic targets in schizophrenia. AB - Evidence implicating dysfunction of glutamatergic neurotransmission rests largely on the finding that antagonists of the NMDA subtype of glutamate receptor, especially the dissociative anesthetics like ketamine, can reproduce the full range of symptoms as well as the physiologic manifestation of schizophrenia such as hypofrontality, impaired prepulse inhibition and enhanced subcortical dopamine release. To test the hypothesis that schizophrenia may result from NMDA receptor hypofunction a number of clinical trials have examined the effects of agents that act on the glycine modulatory site on the NMDA receptor. Glycine, D-serine, and the partial agonist, D-cycloserine, have been shown to improve cognition and decrease negative symptoms in schizophrenic subjects receiving typical antipsychotics. Results with D-cycloserine suggest that clozapine may enhance glycine modulatory site occupancy. Preliminary results with an allosteric modulator of the AMPA subtype of glutamate receptor suggest enhanced cognitive functions in subjects treated with clozapine. PMID- 12769627 TI - N-methyl-D-aspartate receptor-coupled glycineB receptors in the pathogenesis and treatment of schizophrenia: a critical review. AB - Glutamatergic pathways, metabotropic receptors, and ionotropic alpha-amino-3 hydroxy-5-methylisoxazole-4-proprionic acid (AMPA), kainate and N-methyl-D aspartate (NMDA) receptors are all implicated in the etiology and management of schizophrenia. As concerns NMDA receptors, open channel blockers (OCBs) such as phencyclidine (PCP) elicit psychotic symptoms in human subjects. This observation underpins biochemical studies indicating that a deficit in activity at NMDA receptors may be associated with psychotic states. Inasmuch as agonists at the NMDA recognition site are excitotoxic, drugs acting via the co-agonist, glycine(B) (GLY(B)) site are more promising clinical candidates as antipsychotic agents. Glycine (GLY) itself, a further endogenous agonist, D-Serine, and inhibitors of GLY reuptake are active in certain experimental models predictive of antipsychotic properties. Further, in controlled clinical trials, GLY and D Serine enhance the ability of conventional neuroleptics such as haloperidol to improve cognitive and negative symptoms. Their actions are mimicked by the partial agonist, D-cycloserine (DCS). However, these agents exert little effect alone and may interfere with therapeutic actions of the atypical antipsychotic, clozapine. An important issue in the interpretation of drug actions at GLY(B) sites is their degree of occupation by endogenous GLY and D-Serine - although they are unlikely to be saturated. Further, distinct "subtypes" of GLY(B) site bearing NMDA receptor may fulfill differential roles in psychotic states Finally, blockade of certain populations of NMDA receptor may be of use in the management of schizophrenia. This article reviews the complex role of GLY(B) sites/NMDA receptors and their endogenous ligands in the pathogenesis and treatment of psychotic states. PMID- 12769628 TI - Preclinical pharmacology of mGlu2/3 receptor agonists: novel agents for schizophrenia? AB - Agonists for mGlu2/3 receptors decrease the evoked release of glutamate at certain (ie. forebrain / limbic) glutamatergic synapses, indicating that the functional role of mGlu2 and/or mGlu3 receptors is to suppress glutamate excitations. This offers a mechanism for dampening glutamate excitation under pathological states resulting from excessive glutamate release. Based, in part, on the psychotomimetic actions of phencyclidine (PCP)- like drugs, excessive or pathological glutamate release has been implicated in a number of clinical conditions including psychosis. With this in mind, the pharmacology of multiple mGlu2/3 receptor agonists have been investigated in PCP treated rats. Agonists for mGlu2/3 receptors such as LY354740 and LY379268 have been shown to block certain behavioral responses to PCP in rats. The effects of mGlu2/3 agonists on PCP-induced behaviors are blocked by a low doses of a selective mGlu2/3 receptor antagonist, indicating that these actions are mediated via mGlu2/3 receptors. In addition, mGlu2/3 agonists potently suppress glutamate release in rat prefrontal cortex, as reflected by excitatory post-synaptic potentials (EPSPs) induced by serotonin (5-HT) acting on 5HT(2A) receptors. These actions of LY354740 and LY379268 are also blocked by a selective mGlu2/3 antagonist. Atypical antipsychotic drugs such as clozapine also suppress 5-HT-induced EPSPs in this brain region, thus suggesting a common pathway for the actions of atypical antipsychotic drugs and mGlu2/3 receptor agonists. As glutamatergic dysfunction has been implicated in psychotic states and possibly in the etiology of schizophrenia, clinical studies with mGlu2/3 agonists may be warranted to further explore the validity of the glutamatergic hypothesis of schizophrenia. PMID- 12769630 TI - Current status of antipsychotic treatment. AB - With the use of chlorpromazine and other traditional antipsychotics for psychosis, it was soon discovered that the antipsychotic efficacy of this class of medications was closely associated with their ability to block dopamine D(2) receptors in the brain. This prompted the hypothesis that the etiology of schizophrenia and other psychotic illnesses might be caused by a dysregulation of dopamine. This hypothesis, that the dopamine system explains schizophrenia symptoms, however, is far from complete and the treatment with conventional antipsychotic medications is far from ideal. There has been a great deal of speculation regarding the role of serotonin receptor antagonism in regards to antipsychotic effects. The second generation antipsychotics (SGAs), clozapine, risperidone, olanzapine, quetiapine and ziprasidone all have relatively high serotonin to dopamine binding ratios. Serotonin receptor binding may be important to these drugs' actions, possibly by stimulating dopamine activity in mesocortical pathways. Yet, while the mechanism of action of SGAs as a group remain unsolved, it is important to note that the SGAs offer many clinical benefits to treatment as compared to traditional antipsychotics and are quickly emerging as first-line therapy for schizophrenia. In addition to lower rates of EPS and tardive dyskinesia, other benefits to treatment with this class of antipsychotics include better treatment of negative symptoms, better compliance, possible benefits for cognitive impairments, lower rates of relapse and rehospitalization, and more cost-effective therapy. Within the class of SGAs, however, differences exist both in efficacy and side effects and these will be described. Optimization of treatment and understanding the exact mechanism of action of current antipsychotic medications will help pave the way for new drug targets in the future. PMID- 12769629 TI - Novel directions in antipsychotic target identification using gene arrays. AB - Schizophrenia is a major health problem that affects 2 million individuals in the United States. Antipsychotics offer considerable symptomatic relief and, although commonly discovered by screening with single biological targets, most interact with multiple receptors and signaling pathways. Considerable evidence from family and twin studies demonstrates genetic components and multiple chromosomal regions associated with schizophrenia. The polygenic nature of schizophrenia and multiple mechanisms for most effective agents indicate the need for broader approaches to target identification. Gene expression profiling of post-mortem human brain tissue simultaneously reveals the expression of many thousands of genes. A comparison of tissue from normals and patients provides a 'disease signature' of aberrantly expressed genes. 'Drug signatures' are the gene expression changes of cultured human or animal neurons treated with psychiatric drugs, and from animals chronically treated with these drugs. A selection of genes from disease and drug signatures can create a set of targets whose changes may better predict disease and its treatment by effective agents. This multi-parameter high throughput screening (MPHTS(SM)) approach evaluates the mRNA expression pattern of cultured cells exposed to candidate compounds. Compounds that normalize genes altered in schizophrenia may better address its underlying causes. Drugs that mimic gene expression changes that are consistently altered by effective antipsychotic agents provide a drug improvement strategy if efficacy is enhanced or side effects are attenuated. PMID- 12769631 TI - Direct and indirect modulation of the N-methyl D-aspartate receptor. AB - The dopamine hypothesis of schizophrenia has been a driving force in guiding both theories of pathophysiology of schizophrenia, and drug discovery efforts in this area. While this path has been fruitful in producing a deeper understanding of the disorder and a variety of antipsychotic drugs, it is generally recognized that targeting the D(2) dopamine receptor has multiple shortcomings. Recently, alterations in the glutamatergic system have been proposed to play a key role in the neurochemical disruptions underlying schizophrenia. In particular, the similarities between the symptoms of schizophrenia and the psychotomimetic effects of non-competitive N-methyl D-aspartate (NMDA) receptor antagonists such as phencyclidine have spurred interest in the possibility that an NMDA receptor hypofunctional state might underlie schizophrenia. In this review, we summarize the NMDA hypofunction hypothesis of schizophrenia, and focus on the NMDA receptor as a potential target for novel antipsychotic agents. Both modulatory sites on the NMDA receptor, as well as G-protein coupled receptors such as the muscarinic and metabotropic glutamate receptors that modulate the NMDA receptor, are potential targets for the development of novel compounds that could ameliorate the symptoms of schizophrenia. PMID- 12769632 TI - Multiple sclerosis: emerging opportunities for therapeutic intervention. AB - Multiple sclerosis (MS) is a demyelinating disease of the central nervous system of unknown etiology. Two of the major therapies for the treatment of MS, interferon-beta and glatiramer acetate, show only limited evidence that long-term treatment slows disability. There is a great need for new drugs that will halt, reverse, and prevent the development of MS. This article reviews therapies currently in use and describes innovative strategies being developed to alter the disease course. New technologies in gene expression profiling offer hopeful directions toward the design of successful drug therapies and diagnostic testing for MS. Additionally, the new fields of genomics and proteomics offer the promise of novel treatments, and should help to reveal the mechanisms of disease initiation and pathological progression. PMID- 12769633 TI - Targeting the JNK pathway for therapeutic benefit in CNS disease. AB - The c-Jun N-terminal Kinase (JNK) pathway leading to c-Jun phosphorylation plays a causal role in apoptosis of isolated primary embryonic neurons and of multiple neuronal cell lines following a wide variety of stimuli. Activation of this pathway may also contribute to the neuronal atrophy and death that is associated with neurodegenerative pathological conditions including Alzheimer's, Parkinson's, Huntington's Diseases and stroke. Here, the data that providelinks between the activation of the JNK pathway and its potential to play an operative role in CNS disease are reviewed. Also included is the progress on development of inhibitors targeting the JNK pathway for therapeutic benefit. PMID- 12769634 TI - Oiling the pipeline: the path from genomics to drug discovery for brain disease. AB - The advent of genomic technologies has enabled the discovery of key processes involved in the pathogenesis of many disease states. The application of high throughput transcriptional profiling methods holds promise for similar advances in neuroscience; however, particular considerations need to be addressed when initiating these studies in the context of CNS disorders. This review provides a comparison of current genomic technologies, highlighting specific challenges that arise in genomic analysis of brain function and disease, and describes how these approaches have been successfully applied to identify targets for therapeutic intervention. PMID- 12769635 TI - Small molecule Trk receptor agonists and other neurotrophic factor mimetics. AB - Nerve growth factor belongs to a small family of proteins whose binding at the Trk and p75(NTR) transmembrane receptors triggers a cascade of signaling events that give rise to neurotrophic responses in neuronal cells and in vivo. Following their robust effects in animal models of neurodegeneration, neurotrophins have been evaluated for therapy for several human neurodegenerative diseases. However, due mainly to the poor pharmacokinetic behavior of these proteins, they have largely met without success in the clinic, making it desirable to develop small molecule neurotrophin mimetics. A range of compounds is described that achieves some of the neurotrophic and neuroprotective effects attributed to neurotrophins through a variety of mechanisms. These small molecules are divided into the following functional categories: (1). compounds that activate Trk receptors directly; (2). compounds that potentiate the actions of neurotrophins on Trk receptors; (3). compounds that activate Trk indirectly; (4). compounds that influence neurotrophin expression or secretion; and (5). a broad class of compounds that act downstream of, or independently of, Trk receptors. Unfortunately, most of the compounds that have been reported suffer from either lack of specificity for the desired mechanism/effect(s) or lack of efficacy of the compounds in appropriate in vivo models, or both. This second limitation has been particularly severe for compounds designed to mimic the neurotrophins in their interaction with Trk receptors, an ongoing and formidable challenge. Nevertheless, a small subset of the compounds, acting on intracellular signaling pathways downstream of Trk receptors, shows promise for the future treatment of neurodegenerative diseases. PMID- 12769636 TI - Recent advances in the modulation of voltage-gated ion channels for the treatment of epilepsy. AB - Regardless of the voltage-gated ion channel that is targeted in a drug discovery effort for the treatment of epilepsy, two routes have been followed historically: 1). a compound initially, and often surreptitiously, discovered due to activity in animal seizure models is further optimized by medicinal chemistry, or 2). a molecular target is identified based on the phenotype of transgenic animals, or linkage studies from humans with the disease, and compounds are then investigated within a mechanistic framework. Antagonists of voltage-gated sodium channels have been pursued utilizing primarily the first approach; many of these compounds also have significant activity at other ion channels. Both approaches have been utilized to discover voltage-gated calcium channel antagonists, although most efforts to date have used the first approach. Several spontaneous mutant mice and transgenic animals have been utilized to probe the role of the numerous voltage gated calcium channel subunits and their isoforms as potential molecular targets. Compounds that open or prolong the opening of voltage-gated potassium channels have been discovered using the first approach, with a detailed understanding of the molecular target and mechanism of action coming to light several years later. Genetic evidence from humans is limited to relatively rare forms of epilepsy, and transgenic animals with interesting phenotypes do not always translate into good molecular targets in humans. No clinically-useful antiepileptic drug (AED) has been developed to date that specifically interacts with one, or even one class, of ion channels to produce a therapeutic effect. The tools now exist to search for potent, selective, and safe ion channel modulators for the treatment of epilepsy. This review seeks to summarize the most recent pre-clinical and clinical efforts focused on voltage-gated ion-channels for the development of AEDs. PMID- 12769637 TI - Constitutively activated G protein-coupled receptors: a novel approach to CNS drug discovery. AB - G protein-coupled receptors (GPCRs) represent a major class of signal transduction proteins that modulate various biological functions. GPCRs are one of the most common targets for drug development-currently, 39 of the top 100 marketed drugs in use act directly or indirectly through activation or blockade of GPCR-mediated receptors. Nearly 160 GPCRs have been identified based on their gene sequence and their ability to interact with known endogenous ligands. However, an estimated 500-800 additional GPCRs have been classified as "orphan" receptors (oGPCRs) because their endogenous ligands have not yet been identified. Given that known GPCRs have proven to be such clinically useful drug targets, these oGPCRs represent a rich group of receptor targets for the development of novel and improved medicines. To develop ligands for these potential drug targets requires the ability to identify groups or pools of GPCRs that are likely to be involved in a specific disease process (obesity, schizophrenia, depression, etc.) and to dissect out the pharmacological and signal transduction differences between these GPCR subtypes. It also requires the development of assays to detect ligands of GPCRs even when the endogenous ligands are unidentified. This paper will review novel strategies to identify clinically interesting oGPCRs and to screen for small molecules that act as ligands without prior knowledge of endogenous ligands. This involves the use of constitutively activated GPCRs, a technology that provides a unique opportunity to identify several classes of pharmacological agents, including agonists, inverse agonists and allosteric modulators. PMID- 12769638 TI - Novel and emerging therapies in cardiology and haematology. AB - Reviewing advances in cardiology and haematology together may appear at first sight to require some artificiality to make a satisfying fit. For two reasons, at least, this is not the case. Firstly, convergence in biology has become very clear over the past decade and this could not be better illustrated by the demonstration that the haemangioblast is the common progenitor of both haemapoietic stem cells and vascular endothelium. This opens the way to common (and differential) approaches to the manipulation of these cells, a field at present in its infancy. A second convergence is the common goal of understanding the processes resulting in haemostasis, thrombosis and vascular occlusion and the means for developing effective antithrombotics. This is exemplified by a number of agents either in use or in clinical trial as a result of haematological and cardiological collaboration. This collaboration is recognisable with the development, many years ago, of streptokinase and the use of aspirin in vascular disease and continues to this day with specific antiplatelet inhibitors and oral thrombin inhibitors as they become accepted into clinical use over the next few years. Here we review current advances in pharmacological treatments in cardiology and haematology, grouped primarily by disease process, focusing on novel and emerging therapies likely to be of importance in the future. PMID- 12769639 TI - Inhibition of platelet adhesion to collagen as a new target for antithrombotic drugs. AB - Platelet adhesion to a damaged blood vessel is the initial trigger for arterial hemostasis and thrombosis. Platelets adhere to the subendothelium through an interaction with von Willebrand factor (VWF), which forms a bridge between collagen within the damaged vessel wall and the platelet receptor glycoprotein Ib/V/IX (GPIb), an interaction especially important under high shear conditions[1]. This reversible adhesion allows platelets to roll over the damaged area, which is then followed by a firm adhesion mediated by the collagen receptors (alpha(2)beta(1), GPVI, ) in addition[2] resulting in platelet activation. This leads to the conformational activation of the platelet alpha(IIb)beta3 receptor, fibrinogen binding and finally to platelet aggregation. Over the past decades, modulation of platelet function has been a strategy for the control of cardiovascular disease. Lately, drugs have been developed that target the fibrinogen receptor alphaIIbbeta3 or the ADP receptor and many of these promising compounds have been tested in clinical trials. However the development of products that interfere with the first step of hemostasis, i.e. the platelet adhesion, has lagged behind. In this review we want to discuss (i) the in vivo studies that were performed with compounds that target proteins involved in different adhesion steps i.e. the VWF-GPIb-axis, the collagen-VWF axis and the collagen-collagen receptor axis and (ii) the possible advantages these putative new drugs could have over the current antiplatelet agents. PMID- 12769641 TI - Observations on the use of the avian chorioallantoic membrane (CAM) model in investigations into angiogenesis. AB - The chorioallantoic membrane (CAM) is widely used as a model to examine angiogenesis, and anti-angiogenesis. Its advantages over mammalian systems include low cost, and ease of preparation, as well as the absence of a mature immune system. Although the use of this model presents a major opportunity to compare data generated in different laboratories, thereby expediting the evaluation of new drugs, angiogenic potential of cells and tissues, and body fluids, as well as to provide meaningful information concerning the molecular mechanisms involved, the wide range of methodologies used, especially in the quantification of the response, make any comparison essentially invalid. In this review, the major methodologies for all aspects of the use of the CAM in angiogenesis-related studies have been described. These include the source of the CAM, the methods for culture, and methods for evaluation of normal growth and of the response to an intervention. Methods for applying an intervention, the age for intervention and the duration of an intervention are documented. The structure and growth characteristics, the nature of possible responses to stimulation and inhibition of angiogenesis, and the complications of non-specific reactions have been examined. The need for a standardized approach to the use of the CAM model is obvious. One set of possible parameters is suggested. PMID- 12769640 TI - Erythropoietin: cytoprotection in vascular and neuronal cells. AB - One of the principal functions of erythropoietin (EPO) is to stimulate the survival, proliferation, and differentiation of immature erythroid cells. Yet, EPO has recently been shown to modulate cellular signal transduction pathways to perform multiple functions other than erythropoiesis. EPO is cytoprotective through the prevention of programmed cell death in both vascular and neuronal systems by modulating two distinct components of programmed cell death that involve the degradation of genomic DNA and the externalization of cellular membrane phosphatidylserine (PS) residues. Cytoprotection by EPO is initiated by the activation of the EPO receptor (EPOR) and subsequent signal transduction pathways that originate with the Janus-tyrosine kinase 2 (Jak2) protein. Further down-stream cellular pathways include the activation of signal transducers and activators of transcription (STATs), Bcl-x(L), phosphoinositide-3-kinase/Akt, mitogen-activated protein kinases, cysteine proteases, protein tyrosine phosphatases, and nuclear factor kappaB. Further understanding of the cellular pathways that modulate EPO cytoprotection in the nervous system will be crucial for the development of therapeutic strategies against neurodegenerative diseases. PMID- 12769642 TI - Regulatory light chains of striated muscle myosin. Structure, function and malfunction. AB - Striated (skeletal and cardiac) muscle is activated by the binding of Ca(2+) to troponin C and is regulated by the thin filament proteins, tropomyosin and troponin. Unlike in molluscan or smooth muscles, the myosin regulatory light chains (RLC) of striated muscles do not play a major regulatory role and their function is still not well understood. The N-terminal domain of RLC contains a 'Ca(2+)-Mg(2+)'-binding site and, analogous to that of smooth muscle myosin, also contains a phosphorylation site. During muscle contraction, the increase in Ca(2+) concentration activates the Ca(2+)/calmodulin-dependent myosin light chain kinase and leads to phosphorylation of the RLC. In agreement with other laboratories we have demonstrated that phosphorylation and Ca(2+) binding to the RLC play an important modulatory role in striated muscle contraction. Furthermore, the ventricular isoform of human cardiac RLC has been shown to be one of the sarcomeric proteins associated with familial hypertrophic cardiomyopathy (FHC), an autosomal dominant disease characterized by left ventricular hypertrophy, myofibrillar disarray and sudden cardiac death. Our recent studies have demonstrated that phosphorylation and Ca(2+) binding to human ventricular RLC are significantly altered by the FHC mutations and that their detrimental effects depend upon the specific position of the missense mutation, whether located in the proximity of the RLC 'Ca(2+)-Mg(2+)'-binding site or the phosphorylation site (Serine 15). We have also shown that there is a functional coupling between Ca(2+) and/or Mg(2+) binding to the RLC and phosphorylation and that the FHC mutations can affect this relationship. Further in vivo studies are necessary to investigate the mechanisms involved in the pathogenesis of RLC linked FHC. PMID- 12769643 TI - Remodeling of the myocardium and potential targets in the collagen degradation and synthesis pathways. AB - Remodeling of the myocardium is the major mechanism for disability and death in prevalent cardiovascular diseases such as hypertension, heart failure and myocardial infarction (MI). It is a complex process that involves changes in structure, shape and topography at the global level and changes in myocytes and non-myocytes at cellular and subcellular levels that impact negatively on function. Although the myocytes subserve the heart's pump function, the predominant cell type in the heart is the fibroblast (not the myocyte). The fibroblast's major role is deposition of the extracellular matrix (ECM) of which collagen is the principal component. The cardiac extracellular collagen matrix (ECCM) maintains structural and functional integrity, and contributes to coordinated mechanical action with every systole during life. Excessive collagen deposition or pathological fibrosis is an important contributor to left ventricular (LV) dysfunction and poor outcome in hypertension, MI and heart failure. It is also an important problem in the aging heart. Antifibrotic agents that target steps in the collagen synthesis and degradation pathways therefore represent promising strategies for these diseases. Because reparative fibrosis is an essential component of healing of the infarct zone (IZ) after MI, the design of approaches that separately target the IZ and non-infarct zone (NIZ) is challenging. It may be possible in future to target the collagen pathways in the heart or regions of the heart, and not other areas or organs, by delivering drugs or genes locally to specific regions. PMID- 12769644 TI - Retinoids: pleiotropic agents of therapy for vascular diseases? AB - Retinoids, the natural and synthetic derivatives of vitamin A, exert broad biological effects and have been used clinically to treat a variety of dermatological and neoplastic diseases. The principal mode of action of many retinoids is through the binding and activation of a family of nuclear receptors that modulate gene transcription. Recent evidence demonstrates that retinoids effectively attenuate experimental vessel wall narrowing due to atherosclerosis, post-balloon injury stenosis, and bypass graft failure. Moreover, retinoids promote a differentiated phenotype in smooth muscle cells (SMC) which, unlike other muscle types, is not fixed and is subject to considerable modulation in disease states. A growing number of in vitro studies have reported desirable effects of retinoids on cell migration, proliferation, apoptosis, matrix remodeling, fibrinolysis, coagulation, and inflammation, all of which impinge on vascular disease. Since vascular SMC and endothelial cells (EC) express most retinoid receptors, the mechanisms underlying retinoid-mediated events in these cells and the vessel wall likely relate to an altered transcriptome. In fact, there is a growing list of retinoid-response genes encoding proteins that likely mediate the actions of retinoids. Retinoid-response genes, therefore, represent promising targets of therapy for the refined treatment of vascular diseases. The purpose of this review is to summarize the emerging importance of retinoids in the control of vascular cell responses with special emphasis on potential mechanisms underlying retinoid-induced changes in the vessel wall following injury. Given the similarities in the pathogenesis of neoplasia and vascular disease, it is reasonable to consider testing the efficacy of retinoids for the treatment of human vascular disease. PMID- 12769645 TI - New target molecules in the drug control of blood pressure and circulation. AB - Ion channels play a pivotal role in blood pressure regulation. Amongst them, much attention has been directed to dihydropyridine (DHP)-sensitive (L-type) voltage dependent Ca(2+) channels (VDCCs) and iberiotoxin-sensitive Ca(2+)-dependent K(+) channels which are distributed over the whole vascular tree and contribute to vascular tone regulation. Recent advances in vascular electrophysiology have, however, added novel and interesting molecules to this repertoire. In small mesenteric arterioles, the predominant VDCC phenotype is not L-type but DHP insensitive, high voltage-activated VDCCs that exhibit unique properties distinguishable from those of hitherto-known VDCCs. Surprisingly, mibefradil, a well-known T-type selective blocker potently inhibits these channels, and the use of this blocker has indicated that Ca(2+) entry through these channels may be one of the important determinants of peripheral vascular tone. Another new candidate likely involved in blood pressure control is the mammalian homologue of Drosophila transient receptor potential (TRP) protein, including TRPC4 and TRPC6. Experiments in genetically engineered TRPC4-deficient mice have suggested that expression of TRPC4 is indispensable for agonist-induced Ca(2+) entry in endothelial cells and production of nitric oxide and vasorelaxation. TRPC6 is likely to contribute to sustained Ca(2+) entry into vascular smooth muscle cells activated by stimulation of sympathetic nerves and elevation of intravascular pressure. Antisense oligonucleotide experiments have suggested that this protein is an essential component of alpha1-adrenoceptor activated and mechanosensitive cation channels in some vascular tissues. This review overviews what is known about the role of ionic channels in blood pressure control with main focus on the above-mentioned new molecules as promising targets for drug discovery and development. PMID- 12769646 TI - RAS blockade in experimental renal transplantation. Benefits and limitations. AB - In renal transplantation, chronic renal transplant failure (CRTF) is the principal cause of late graft loss. Both immunological and non-immunological factors play a role in the pathogenesis of CRTF. However, CRTF is unresponsive to immunosuppressive therapy. In several kidney diseases, inhibition of the renin angiotensin system (RAS) has shown to reduce the rate of progression of renal disease more effectively than conventional antihypertensive drugs. Therefore, RAS blockade may be of benefit in the treatment of CRTF. Several short-term studies in human renal transplant recipients showed that RAS blockade had a beneficial effect on renal transplant function, blood pressure and proteinuria. Despite these benefits physicians remain reluctant to use ACE inhibition in these recipients, because of fear of functional decrease in renal perfusion, especially in the setting of renal transplant artery stenosis. To study the long-term effects of RAS blockade we used the established Fisher to Lewis (F-L) model for CRTF, which mirrors the progressive changes seen in humans. Studies in our lab and by others showed that RAS blockade in the F-L model prevents proteinuria, glomerulosclerosis and hypertension. However, when treated for 34 weeks with RAS blockade, renal arteries developed severe intimal hyperplasia. This effect was specific for Fisher rats. Syngrafted Fisher rats treated with ACE inhibition developed intimal hyperplasia, but allografting significantly aggravated it. Fisher rats have a four times higher renal ACE activity, compared with the Lewis rat. This is comparable to the human DD/II genotype differences in ACE activity. Renal transplant patients with the DD genotype may be more vulnerable for vascular changes when treated with RAS blockade. PMID- 12769647 TI - Viral and cellular cytokines as therapeutic targets in AIDS-related lymphoproliferative disorders. AB - Since the advent of highly active antiretroviral therapy (HAART) and its widespread use, the incidence of AIDS-defining illnesses has decreased dramatically, leading to a much longer survival of patients. Despite some exciting new leads, non-Hodgkin's lymphoma (NHL) remains a fatal malignancy for the vast majority of patients with acquired immunodeficiency syndrome (AIDS). Multiple molecular pathways appear to operate in AIDS lymphomagenesis and some may preferentially be associated with specific malignant histopathologic categories or anatomic sites of origin. AIDS-associated lymphomas share several features, including B-cell lineage derivation, diffuse aggressive histology, and frequent origin from or involvement of extranodal sites. Recently, high-grade primary effusion lymphomas (PEL) have been reported in patients with advanced AIDS. PEL is recognized as a distinct clinicopathologic entity associated with Kaposi's sarcoma-associated herpesvirus (KSHV), also known as human herpesvirus-8 (HHV-8). KSHV genes are likely to contribute to the neoplastic phenotype of PEL cells that require cytokines and factors from the host or encoded by the virus for growth in vivo. KSHV is also thought to dramatically affect the incidence, type, and course of multicentric Castleman's disease, another lymphoproliferative disorder over-represented in patients with AIDS. This review summarizes the current knowledge of autocrine growth factor loops and angiogenic factors that are involved in the pathogenesis of KSHV-related lymphoproliferative disorders in AIDS. Deregulated cytokines may represent potential targets of novel therapeutic strategies. PMID- 12769648 TI - Small molecule immunosuppressive agents in experimental and clinical transplantation. AB - Immunosuppression is currently the major approach used for the prevention and management of transplant rejection. In this overview, preclinical and clinical studies of the small molecule immunosuppressive agents are reviewed from the discovery of cyclosporine. More recently it was demonstrated that certain agents, namely tacrolimus (FK506), sirolimus (rapamycin), and mycophenolate mofetil (CellCept, MMF), act selectively on adaptive host responses at different stages of T- and B-cell cycles and spare nonspecific host resistance. Because each agent has its specific and significant toxic effects, it has been difficult to optimize the use of individual agents in monotherapy. Therefore, drug combination therapy has been of great interest in addition to the introduction of new small molecule agents, such as malononitrilamides [MNAs (leflunomide, FK778, FK779)], 15 deoxyspergualin (DSG) and its analogues, FTY720 and inhibitors of signal transduction, which offer promising modes of immunosuppression. PMID- 12769649 TI - New immunosuppressants in clinical trial. AB - Chemical immunosuppression developed at the beginning of clinical transplantation remains the basis of immunosuppressive protocols used today. Clinical trials of new agents focus on the incremental improvement in one of four areas of immunotherapy: induction, basic maintenance, adjuvant and steroid therapy. Induction immunotherapy developments include the use of humanized antibodies, specific targets (basiliximab, daclizumab) and lymphocyte depletion (anti thymocyte globulin, campath-1H). Promising results from pilot trials allow for subsequent low dose maintenance monotherapy. New inhibitors of calcineurin isoforms (ISATx247) may disassociate the anti-rejection effect from the toxicity of conventional calcineurin inhibitors. Revived investigation of older anti proliferative or anti-metabolite drugs (rapamycin, mycophenolic acid, leflunomide) show promise in preventing B-cell responses and reducing chronic rejection but development of derivatives (everolimus, mycophenolate mofetil, ERL 080, MNA 279, MNA 715) may have a stronger commercial than experimental basis. Trials of steroid immunotherapy have focused on steroid elimination but trial of newer locally active steroids may be beneficial. PMID- 12769650 TI - Pharmacotherapeutic approach to prevent or treat chronic allograft nephropathy. AB - In the current paper we will review the evidence that drug therapy may be of value to prevent or treat chronic allograft nephropathy. We will review the immunosuppressive therapy and non-immune therapies in use to treat risk factors associated with chronic allograft nephropathy and evaluate their efficacy with respect to long term outcome as well as their effect on markers of long-term survival. In the last part of this review, we will indicate possible benefits of new approaches being explored but most of these data are obtained in in vitro test systems and rodent models. PMID- 12769651 TI - Novel immunosuppressive agents in tolerance induction. AB - Induction of transplantation tolerance remains a much sought-after but elusive goal with a potential promise of rendering patients free of long-term immunosuppressive drugs while maintaining good organ transplant function indefinitely. All currently studied strategies toward transplantation tolerance include using immunosuppressive agents for a limited time period to achieve tolerance. There are a growing number of agents that lend themselves to tolerance induction and these agents are considered herein. Such agents generally fall into the categories of lymphocyte depletion, costimulation blockade, and adjunctive agents such as rapamycin. Preliminary data using donor stem cells suggests that embryonic stem cells may have substantial advantages over conventional donor bone marrow for tolerance induction. These preliminary results are briefly reviewed as well. Tolerance has been achieved in a small number of organ transplant patients and such successes suggest that progress in the clinical arena is happening and is an obtainable result. PMID- 12769652 TI - Therapeutic interventions in xenotransplantation. AB - Xenotransplantation, involving the transplantation of pig organs into humans, would resolve the current shortage of organs. It involves, however, a new therapeutic approach to organ transplantation. The presence of natural antibody in primates directed against Galalpha1,3Gal epitopes on pig vascular endothelium leads to early antibody-mediated rejection. An elicited antibody response against the same target epitopes as well as against nonGal antigens intensifies the immune destruction of the organ. Even the minimal deposition of antibody appears to lead to the development of a consumptive coagulopathy that can be fatal. Approaches being investigated to overcome these barriers include depletion and inhibition of natural antibody and complement, and suppression of the elicited T cell-dependent antibody and cellular responses. In addition, however, physiologic incompatibilities between human and pig, particularly those relating to coagulation, may enhance or complicate the immune process, and may require additional therapeutic measures. Current approaches aimed at achieving successful xenotransplantation also include investigation of agents that prevent potential xenozoonotic infection of the recipient. At present, therefore, the therapeutic interventions required to attempt to overcome the barriers to xenotransplantation are multiple. Work indicating progress in the breeding of pigs that do not express the critical Galalpha1,3Gal epitopes, however, is encouraging. The introduction of these pigs may greatly reduce the therapy required, and may ultimately allow the development of methods to induce tolerance to the transplanted pig organ. PMID- 12769653 TI - Ca2+-dependent signaling pathways in the heart: potential drug targets for cardiac disease. AB - Ca2+ is an important secondary messenger and any alteration to intracellular Ca2+ signaling pathways or components of these pathways can have a profound physiological effect on any cell, particularly cardiomyocytes. Early approaches to investigate heart disease focused on many muscle proteins, however recent findings indicate that molecules considered as "non-muscle" proteins may be equally important players in etiology of many cardiac pathologies. Many of these "non-muscle" proteins play a role in Ca2+ cycling or Ca2+ -dependent signaling pathways in the heart. In this review we focus on Ca2+ -dependent pathways in normal, growing, and diseased hearts. Understanding of these unique signaling pathways may hold answers to many cardiac pathologies in children and adults. PMID- 12769654 TI - The antithrombotic efficacy of lotrafiban (SB 214857) in canine models of acute coronary thrombosis. AB - In patients with acute coronary syndromes, inhibition of platelet aggregation with parenteral alpha(IIb)/beta(III) antagonists has proven effective at preventing nonfatal myocardial infarction and repeat percutaneous coronary interventions. Paradoxically, the efficacy observed for acute indications and parenteral agents has not extended to oral agents and chronic prevention of secondary thrombotic events, despite robust antithrombotic properties in preclinical thrombosis models. This report documents the preclinical data of Lotrafiban, an oral alpha(IIb)/beta(III) antagonist that recently failed in a phase III clinical trial (BRAVO) for the prevention of secondary thrombosis. Lotrafiban was characterized in a dog circumflex artery electrical injury model, and a cyclic flow reduction model ("Folts"). The data demonstrate that both oral (1.0-50.0 mg/kg) and intravenous (0.1-0.8ug/kg/min) administration of lotrafiban produced dose-related inhibition (45%-95%) of ex vivo platelet aggregation. In the electrical injury model, the dose-related inhibition correlated with a significant reduction in the frequency of coronary occlusion, size of the developing thrombus, and the extent of left ventricular ischemic damage. Effects on blood flow and bleeding time were also dose related. The combination of low dose lotrafiban (0.1ug/kg/min) and aspirin (5.0 mg/kg) generated additive antithrombotic effects, approximating the antithrombotic efficacy of a 2-4 fold higher dose of lotrafiban while only modestly prolonging the bleeding time. For purposes of comparison, the ADP receptor antagonist clopidogrel was also assessed in the electrical injury model. Clopidogrel (5.0-10.0 mg/kg, iv.) significantly reduced the resulting left ventricular infarct areas, but lacked the overall efficacy of lotrafiban. In the "Folts" model, lotrafiban inhibited cyclic blood flow reductions (CFR's) by 100% in animals insensitive to the antithrombotic effects of aspirin. Overall, the preclinical data demonstrated that alpha(IIb)/beta(III) antagonism with lotrafiban was a well tolerated and effective strategy for attenuating acute arterial thrombosis. The lack of a correlation between these preclinical data and the outcome of the clinical trial BRAVO are unexplained. However, the combined evidence suggests that these acute canine thrombosis studies may not completely capture the pathology reflected in chronic human atherothrombotic disease. PMID- 12769655 TI - Inhibition of PAI-1: a new anti-thrombotic approach. AB - Proteolytic degradation of fibrin (fibrinolysis) is mediated by plasminogen and its activators, tissue-type plasminogen activator (tPA(1)) and urokinase (uPA). Fibrinolysis is critical for preventing thrombus growth and restoring blood flow following thrombotic vascular occlusion. Plasminogen activator inhibitor-1 (PAI 1), a member of the serine protease inhibitor (serpin) superfamily, is the principal inhibitor of tPA and uPA in the fibrinolytic system. High levels of circulating PAI-1 are associated with a number of thrombotic diseases. In animal studies, transgenic mice overexpressing human PAI-1 develop spontaneous thrombosis, whereas PAI-1-deficient mice are more resistant to venous or arterial thrombosis. Furthermore, inhibition of PAI-1 activity prevents thrombus formation in animal models. The antithrombotic effects of PAI-1 inhibition are achieved by enhancing endogenous fibrinolytic activity without directly affecting blood coagulation and platelet function. Phenotypic analysis of PAI-1 deficiency in both human and mouse suggests that inhibition of PAI-1 will not lead to severe bleeding or other major adverse effects. Thus, PAI-1 inhibitors represent a new class of antithrombotic drugs with a possible wider therapeutic index than conventional antiplatelet and anticoagulant agents. This review summarizes the role of PAI-1 in thrombotic diseases and recent progress in the development of small molecule PAI-1 inhibitors. PMID- 12769656 TI - Trypanosoma cruzi trypanothione reductase inhibitors: phenothiazines and related compounds modify experimental Chagas' disease evolution. AB - Chagas' disease affects about 18 million people and 25% of the population of Latin America is at risk of acquiring Chagas' disease. The chemotherapy of Chagas' disease is still an open field and remains as an unsolved problem. Nifurtimox and benznidazole are currently used to treat this disease, however, both drugs have high toxicity and are mutagenic with the result that the patients frequently fail to follow treatment. T. cruzi enzimes such as trypanothione reductase, represent potential drug targets because they play an essential role in the life of this organism. This enzyme has been isolated, purified and studied by X ray crystallography. Phenothiazines and related compounds inhibit trypanothione reductase and a specially favoured fit is a phenothiazine with a 2- substitued with 2- chloro and 2- trifluoromethyl with a remote hydrophobic patch. The essential phenothiazine nucleus can adopt more than one inhibitory orientation in its binding site. Phenothiazines and related compounds are drugs used in psychiatric treatments. These anti-depressants inhibit trypanothione reductase through the peroxidase/ H2O2/ system, and also exert other trypanocidal effects upon epimastigotes and tripomastigotes forms: clomipramine through an anticalmodulin action; trifluopherazine and thioridazine induced disruption of mitochondria and prometazine provoked serious cell membrane disorganization. Clomipramine and thioridazine were also effective in treatment of mice with experimental Chagas' disease, significantly modifying the natural evolution of the infection; cardiac function and survival of infected and treated animals were not different from non infected animals. Phenothiazines and related compounds are promising trypanocidal agents for treatment of Chagas' disease. Other trypanocidal agents as nifurtimox, benznidazol,Allopurinol, cystein protease inhibitors and others, are also discussed. PMID- 12769657 TI - Thrombin activable fibrinolysis inhibitor (TAFI): molecular genetics of an emerging potential risk factor for thrombotic disorders. AB - The balance between the activities of the coagulation and fibrinolytic cascades is crucial for normal hemostasis. However, imbalances can lead to pathological thrombotic events, as is observed in heart attacks and strokes, as well as excessive bleeding, as in hemophilia. Recent investigations have uncovered a novel molecular connection between the two cascades that has been termed thrombin activable fibrinolysis inhibitor (TAFI) as well as procarboxypeptidase U, procarboxypeptidase R or plasma procarboxypeptidase B. TAFI is the precursor of an enzyme (TAFIa) with basic carboxypeptidase activity that attenuates the lysis of fibrin clots by removal of the carboxyl-terminal lysine residues from partially-degraded fibrin that mediate positive feedback in the fibrinolytic cascade. The plasma concentration of TAFI varies substantially (up to approximately 10-fold) in the human population and may constitute a novel risk factor for thrombotic disorders. Sixteen single nucleotide polymorphisms have been identified in the 5'-flanking, protein coding, and 3'-untranslated regions of the TAFI gene. The polymorphisms all have been shown to be associated with variations in plasma TAFI concentrations. One amino acid substitution has been found to directly alter the properties of the TAFIa enzyme. This review provides a general overview of the TAFI pathway, including a discussion of the spectrum of inhibitors of TAFIa that have been described, and summarizes the recent advances in the molecular genetics of the TAFI gene as well as the results of studies that may implicate the TAFI pathway in risk for arterial and venous thrombotic disorders. PMID- 12769659 TI - Apolipoprotein E: possible therapeutic target for atherosclerosis. AB - Human apolipoprotein E (apoE) consists of a single polypeptide chain with 299 amino acids and is best known for its role in the transport of cholesterol and other lipids between peripheral tissues and the liver. However, more direct effects of apoE on the vascular wall may well contribute to arterial protection from atherosclerosis. This review will focus on: (a) the ability of apoE to direct cholesterol efflux mechanisms with the aid of apoA1 and the ATP binding cassette transporter 1 (ABC1); (b) the ability of apoE to prevent platelet aggregation by facilitating the production of endogenous nitric oxide (NO); (c) the ability of apoE to inhibit the proliferation of T-lymphocytes by internalization of the IL-2 receptor; and (d) the ability of apoE to inhibit proliferation of endothelial cells by out competing growth factors for interaction with cell surface heparan sulfate proteoglycans (HSPG's). The characterization of apoE and its many functions has provided insight into the ultimate potential of this protein as a possible therapeutic agent for the treatment of atherosclerosis. This review will examine key scientific advances, which focus on possible therapeutic strategies that encompass the use of apoE in the amelioration of atherosclerosis. PMID- 12769658 TI - Contributory role of matrix metalloproteinases in cardiovascular remodeling. AB - Changes in cardiovascular tissue structure can result in alterations in function. This process occurs as a continuum and can be defined as cardiovascular remodeling. A family of zinc dependent proteases known as the matrix metalloproteinases (MMPs) has been demonstrated to cause tissue remodeling. A number of past studies have demonstrated increased expression and activation of MMPs in both myocardial and vascular remodeling processes. Experimental results support the concept that the MMPs directly contribute to progressive myocardial remodeling post-myocardial infarction and in cardiomyopathic disease. Within the vascular compartment, compelling evidence exists for the contributory role of MMPs in atheromatous plaque rupture and aortic aneurysms. Therefore, the purpose of this review is to examine recent studies of this proteolytic system in the context of cardiovascular remodeling. PMID- 12769660 TI - Reduction and prevention of the cardiovascular sequelae of the insulin resistance syndrome. AB - Insulin resistance and hyperinsulinemia are the critical characteristics of the metabolic syndrome that is associated with abdominal obesity and are the early manifestations of its progression to type 2 diabetes. These metabolic abnormalities are becoming recognized as a major contributor to cardiovascular disease. The experimental studies required to elucidate the underlying mechanisms and to develop effective preventative strategies will require the use of appropriate animal models and these are available. The evidence from such research indicates that a wide range of interventions (including peroxisome proliferator activator receptor agonists, insulin-sensitizing agents, statins, fibrates, angiotensin-converting enzyme inhibitors, estrogen receptor modulators, lipid-based nutriceuticals, and ethanol) can markedly reduce or prevent vasculopathy and ischemic cardiac lesions in animal models. Overall, the results suggest that early damage to the vascular wall, both in function and presenting as atherosclerotic lesions, is secondary to long-term hyperinsulinemia and, especially, to postprandial peaks in plasma insulin levels, and is exacerbated by the accompanying hyperlipidemia. Effective treatment will, of necessity, be preventative and will necessitate diagnostic approaches that can identify asymptomatic individuals at high risk for vascular damage and eventual progression to type 2 diabetes. Therapeutic targets in this population include insulin sensitivity and the associated signal transduction pathways, the peroxisome proliferator activator receptor-alpha and -gamma systems, and the complex pathways leading from acetyl CoA and the citric acid cycle to the synthesis of fatty acid and the storage of triglyceride. These pharmacological approaches offer the prospect of preventing a significant proportion of cardiovascular disease. PMID- 12769661 TI - Modification of clearance of therapeutic and potentially therapeutic proteins. AB - Advances in biochemistry, protein chemistry and molecular biology over the last twenty-five years have spurred the increased use and development of proteins as injectable therapeutic agents. Introduction of proteins into the circulation exposes them to numerous different cells, enzymes and routes of extravasation that contribute to their clearance and their catabolism. Overly rapid clearance, particularly of small proteins, can limit therapeutic efficacy. Many strategies have been devised to retard the clearance of therapeutic or potentially therapeutic proteins, but relatively few proteins with clearance-retarding modifications are in clinical use. Proteins have been chemically modified towards this end by covalent attachment of polyethylene glycol or dextran chains or by protein-protein cross-linking. Genetic modification has also been employed to fuse proteins of interest to long-lived plasma proteins like albumin or immunoglobulins, or portions of these proteins. While all modifications may reduce the biological activity of the protein of interest or elicit antibody formation in recipient animals or patients, there now exists sufficient experience in this area that an optimal clearance-extending strategy can often be designed and successfully executed. With the explosive growth of genomic and proteomic information, an exponentially increasing number of engineered proteins are likely to be developed, with a probable need for clearance-related modification. PMID- 12769662 TI - Cell cycle dependent regulation of intracellular calcium concentration in vascular smooth muscle cells: a potential target for drug therapy. AB - Intracellular Ca2+ transients have been shown to control several transition points within the eukaryotic cell cycle. We focus here on the G1-to-S phase transition triggered by an increase in the intracellular Ca2+ concentration ([Ca2+](i)) in rodent vascular smooth muscle cells (VSMC) and its potential targeting for the treatment of vaso-occlusive processes such as atherosclerosis, hypertension and post-angioplasty restenosis. The transcription factor c-Myb generates a G1/S transition-specific Ca2+ transient via its regulation of a high affinity Ca2+ efflux pump, the plasma membrane Ca2+ ATPase-1 (PMCA1). The cell cycle-associated repression of PMCA1 is mediated by two c-Myb binding sites in the PMCA1 promoter. As c-Myb levels increase in late G1 phase of proliferating VSMC, transcription from the PMCA1 promoter is reduced, expression of the PMCA1 gene falls, and the resultant reduced rate of Ca2+ efflux underlies a G1/S associated increase in [Ca2+](i). Blocking either the upregulation of c-Myb levels, or the down regulation in expression of the efflux pump, leads to significant reductions in S phase entry and proliferation of VSMC. A search for functional c-Myb sites within the promoters of other Ca2+ transporters has been undertaken in order to extend the molecular framework of the G1/S-specific Ca2+ signal mediated by the c-Myb transcription factor. Animal studies with c-myb antisense oligodeoxynucleotides and an anti-c-myb ribozyme as well as in vitro results with dominant negative c-Myb mutants and a doxycycline-inducible c-Myb neutralizing antibody point to the potential of c-Myb-targeted gene therapy for treating pathologic VSMC proliferation and highlight the need for clinical trials in this field. PMID- 12769663 TI - Development and current applications of thrombin-specific inhibitors. AB - Thrombin-specific inhibitors directly diminish thrombin-induced coagulation and cellular activities without the side effects of heparin. Hirudin is the most potent natural thrombin-specific inhibitor. Recombinant hirudins (such as desirudin) have been shown to be effective in the treatment of heparin-induced thrombocytopenia (HIT) and in the prevention of thrombotic complications after hip or knee surgery. The application of recombinant hirudin has been limited mainly by hemorrhagic complications. Synthetic thrombin-specific inhibitors, including oligopeptides, tripeptides and non-peptide low molecular weight (LMW) thrombin inhibitors, have been designed according to their interactions with the active sites of thrombin. Bivalirudin (an anti-thrombin oligopeptide) has been approved for preventing thrombosis in unstable angina patients following angioplasty in adjunct to aspirin. Argotroban (a tripeptide thrombin inhibitor) has been used for the treatment of HIT, peripheral and cerebral thrombotic diseases. The benefit of using thrombin-specific inhibitors alone in acute myocardial infarction or unstable angina remains uncertain. A number of LMW thrombin-specific inhibitors have been developed. Some of them can be administrated orally, and cause less increase in bleeding time than other thrombin inhibitors. The efficacy, safety, stability and oral bioavailability of the thrombin inhibitors may be considerably improved through structural optimization. Most of the LMW thrombin inhibitors are currently being tested in animal models or at early stages of clinical trials. In this review, we will present an overview of recent advances in thrombin-specific inhibitors. PMID- 12769664 TI - Pleiotropic effects of statins on the vascular tissue. AB - The statins are lipid-lowering agents that act by inhibition of 3-hydroxy-3 methylglutaryl-coenzyme A (HMG-CoA) reductase. This enzyme is responsible for the conversion of HMG-CoA to mevalonate. Products of mevalonate metabolism are critical for several cellular processes of eukaryotic cells, and inhibition of the mevalonate pathway by statins has pleiotropic effects. It has been reported that statins inhibit the migration and proliferation of vascular smooth cells (VSMCs), reduce interleukin-6 expression in VSMCs, improve endothelium-dependent vasomotion, and inhibit the expression of plasminogen activator inhibitor-1 and matrix metalloproteinases in endothelial cells. These effects of statins are independent of plasma cholesterol level, and are completely blocked by exogenous mevalonate and some isoprenoids. These findings suggest that statins exert direct antiatherosclerotic effects on the vascular wall beyond their effects on plasma lipids. PMID- 12769665 TI - Drug uptake systems in liver and kidney. AB - The hepatobiliary system and the kidneys are the main routes by which drugs and their metabolites leave the body. Compounds that are mainly excreted into bile in general have relatively high molecular weights, are amphipathic and highly bound to plasma proteins. In contrast, compounds that are predominantly excreted into urine have relatively low molecular weights, are more hydrophilic and generally less protein bound. The first step in drug elimination in liver and kidney is uptake into hepatocytes or into proximal tubular cells. The substrate specificity and affinity of the uptake carriers expressed at the basolateral membranes of hepatocytes and proximal tubular cells could therefore play an important role for the determination of the main elimination route of a compound. This review discusses the tissue distribution, substrate specificity, transport mechanism, and regulation of the members of the organic anion transporting polypeptide (Oatp/OATP) superfamily (solute carrier family SLC21A) and the SLC22A family containing transporters for organic cations (OCTs) and organic anions (OATs). The Oatps/OATPs are mainly important for the hepatic uptake of large amphipathic organic anions, organic cations and uncharged substrates, whereas OCTs and OATs mediate uptake of predominantly small organic cations and anions in liver and kidney. PMID- 12769666 TI - Pharmacokinetics of topical ocular drug delivery: potential uses for the treatment of diseases of the posterior segment and beyond. AB - In developing a drug delivery strategy, issues of absorption, distribution, metabolism, and elimination must be considered. The eye presents unique opportunities and challenges when it comes to the delivery of pharmaceuticals, and is most accessible to the application of topical medications. While absorption by this route is inefficient, there are few side effects. While it has been assumed that topically applied drugs penetrated into the intraocular environment through the cornea, this is currently being reassessed. More recent investigations have shown that the conjunctival route of entry plays an important role in the penetration of drugs into the anterior segment. Furthermore, topically applied drugs have been shown to have access to the sclera from the conjunctiva. As such, it is conceivable that such drugs could find their way to the posterior segment. Data suggest that the sclera is readily permeable to even large molecular weight compounds ( approximately 150 kD). The recent finding that topically applied nepafenac inhibited choroidal and retinal neovascularization by decreasing the production of VEGF, as well as our data showing that even a large molecular weight peptide like insulin can accumulate in the retina and optic nerve after topical application, supports the contention that topically applied drugs can not only reach the posterior segment, but that they can also be therapeutic. Finally, the implications of our findings that topically applied insulin also accumulates in the contralateral eye as well as in the central nervous system are discussed. PMID- 12769667 TI - Tamoxifen: is it safe? Comparison of activation and detoxication mechanisms in rodents and in humans. AB - Tamoxifen, a non-steroidal antiestrogen, is the class representative of a group of drugs that include toremifene, droloxifene and idoxifene. Tamoxifen has been successfully used worldwide as adjuvant therapy in the treatment of women with breast cancer. However, such therapy results in a slightly increased risk of endometrial cancers. Lifetime exposure of rats to high doses of tamoxifen results in a high incidence of liver tumors. Tamoxifen itself is not genotoxic but is activated in the liver to alpha-hydroxytamoxifen. This is further conjugated to form the sulfate ester as the putative reactive intermediate. Studies with recombinant human CYPs show only CYP3A4 is able to catalyze the formation of alpha-hydroxytamoxifen and the irreversible binding of [(14)C]tamoxifen to DNA. CYP3A4 and CYP2D6 convert tamoxifen to N-desmethyltamoxifen. The formation 4 hydroxytamoxifen is catalyzed by CYP2D6 and at a much lower level by CYP2C19. In women, detoxication of alpha-hydroxytamoxifen via a stable glucuronide occurs at a rate in the order of 100 fold higher than in rats whereas rates of sulfation are 3 fold lower than in rats. These factors, together with the low dose of tamoxifen used therapeutically in women, indicates a minimum risk of liver cancers. Results from (32)P-postlabeling and accelerator mass spectrometry suggest that low levels of uterine DNA binding does occur but this is probably too low to play a role in uterine tumor development and it is more likely to be the estrogen agonist action of this class of drug that is the most important factor in tumor development in humans. PMID- 12769668 TI - Gallic acid and gallic acid derivatives: effects on drug metabolizing enzymes. AB - Gallic acid and its structurally related compounds are found widely distributed in fruits and plants. Gallic acid, and its catechin derivatives are also present as one of the main phenolic components of both black and green tea. Esters of gallic acid have a diverse range of industrial uses, as antioxidants in food, in cosmetics and in the pharmaceutical industry. In addition, gallic acid is employed as a source material for inks, paints and colour developers. Studies utilising these compounds have found them to possess many potential therapeutic properties including anti-cancer and antimicrobial properties. In this review, studies of the effects of gallic acid, its esters, and gallic acid catechin derivatives on Phase I and Phase II enzymes are examined. Many published reports of the effects of the in vitro effects of gallic acid and its derivatives on drug metabolising enzymes concern effects directly on substrate (generally drug or mutagen) metabolism or indirectly through observed effects in Ames tests. In the case of the Ames test an antimutagenic effect may be observed through inhibition of CYP activation of indirectly acting mutagens and/or by scavenging of metabolically generated mutagenic electrophiles. There has been considerable interest in the in vivo effects of the gallate esters because of their incorporation into foodstuffs as antioxidants and in the catechin gallates with their potential role as chemoprotective agents. Principally an induction of Phase II enzymes has been observed however more recent studies using HepG2 cells and primary cultures of human hepatocytes provide evidence for the overall complexity of actions of individual components versus complex mixtures, such as those in food. Further systematic studies of mechanisms of induction and inhibition of drug metabolising enzymes by this group of compounds are warranted in the light of their distribution and consequent ingestion, current uses and suggested therapeutic potential. However, it must be noted that numerous constituents of foodstuffs have been found to be potent modulators of xenobiotic metabolism and the net human health effects may depend on concentrations of individual components and individual genetic makeup. PMID- 12769669 TI - Regulation of UDP glucuronosyltransferase genes. AB - The UDP glucuronosyltransferase (UGT) content of cells and tissues is a major determinant of our response to those chemicals that are primarily eliminated by conjugation with glucuronic acid. There are marked interindividual differences in the content of UGTs in the liver and other organs. The mechanisms that lead to these differences are unknown but are most likely the result of differential UGT gene expression. Several transcription factors involved in the regulation of UGT genes have been identified. These include factors such as Hepatocyte Nuclear Factor 1, CAAT-Enhancer Binding Protein, Octamer transcription Factor 1 and Pbx2, which appear to control the constitutive levels of UGTs in tissues and organs. In addition, UGT gene expression is also modulated by hormones, drugs and other foreign chemicals through the action of proteins that bind and/or sense the presence of these chemicals. These proteins include the Ah receptor, members of the nuclear receptor superfamily, such as CAR and PXR and transcription factors that respond to stress. PMID- 12769670 TI - Phenotypic screening of small molecule libraries by high throughput cell imaging. AB - We have developed high throughput fluorescence cell imaging methods to screen chemical libraries for compounds with effects on diverse aspects of cell physiology. We describe screens for compounds that arrest cells in mitosis, that block cell migration, and that block the secretory pathway. Each of these screens yielded specific inhibitors for research use, and the mitosis screen identified Eg5 as a potential target protein for cancer chemotherapy. Cell imaging provides a large amount of information from primary screening data that can be used to distinguish compounds with different effects on cells, and together with automated analysis, to quantitate compound effects. PMID- 12769671 TI - A fluorescent high-throughput assay for double-stranded DNA: the RediPlate PicoGreen assay. AB - The fluorescent PicoGreen reagent for detection and quantitation of double stranded DNA has been adapted for high-throughput screening: the RediPlate PicoGreen double-stranded DNA assay format. In the RediPlate PicoGreen assay format, the PicoGreen reagent is predistributed and co-dried into either 96- or 384-well microplates with the excipient trehalose. The user resuspends the dried reagents upon adding DNA, and measures the resulting fluorescence after a five minute incubation. Replicate fluorescence measurements on nominally identical wells have less than a 5% coefficient of variation. The assay is linear from 5 to 500 ng/ml DNA in a 200 micro l volume. The RediPlate PicoGreen assay format retains the advantages of the original PicoGreen reagent - sensitivity, speed, and specificity - but in a high-throughput format. PMID- 12769672 TI - Rapid assays for quantitating cytokine gene expression without target amplification. AB - Many drug discovery efforts are focused on finding candidates that alter gene expression of the cytokines involved in inflammation, allergy, and cell-mediated immunity. Current methods used to evaluate gene expression such as northern blot and RT-PCR are laborious, time-consuming, expensive, and are not conducive to high throughput screening. High Performance Signal Amplification (HPSA( trade mark )) gene expression assays quantitate mRNA targets directly from cell lysate samples using DNA probe hybridization and fluorescent signal amplification. The assay format eliminates the need for RNA purification prior to testing and does not involve target amplification. The 96 or 384-well microplate formats allow the method to be run manually, by a workstation approach, or with full automation. Cellular mRNA levels are quantitated relative to a standard curve comprised of highly purified in vitro RNA calibrators. The analytical sensitivity is in the low attomole (10(-18) mole) range. This technique was used to monitor the transcription patterns of mRNA encoding TNF-alpha, IL-1beta, and Interferon-gamma in human cell lines or primary PBMC treated with inducers such as PMA, ionomycin, and endotoxin. The specificity, precision and reproducibility of the assay are sufficient to provide a reliable screening system. The HPSA gene expression assay system offers a rapid and convenient alternative to more cumbersome, expensive methods. PMID- 12769673 TI - Assay development and high-throughput screening of caspases in microfluidic format. AB - Caspase proteases are familiar targets in drug discovery. A common format for screening to identify caspase inhibitors employs fluorogenic or colorimetric tetra-peptide substrates in 96, 384, or 1536 -well microtiter plates. The primary motivation for increasing the number of wells per plate is to reduce the reagent cost per test and increase the throughput of HTS operations. There are significant challenges, however, to moving into or beyond the 1536-well format, such as submicroliter liquid handling, liquid evaporation, increased surface area to-volume ratios, and the potential for artifacts and interference from small air borne particles such as lint. Therefore, HTS scientists remain keenly interested in technologies that offer alternatives to the ever-shrinking microtiter plate well. Microfluidic assay technology represents an attractive option that, in theory, consumes only subnanoliter volumes of reagents per test. We have successfully employed a microfluidic assay technology in fluorogenic screening assays for several caspase isoforms utilizing the Caliper Technologies Labchip platform. Caspase-3 is used as a representative case to describe microfluidic assay development and initial high-throughput screening results. In addition, microfluidic screening and plate-based screening are compared in terms of reagent consumption, data quality, and ease of operation. PMID- 12769674 TI - Fluorescence assays for high-throughput screening of protein kinases. AB - Protein kinases comprise one of the most important group of targets for drug discovery research today. Methods to identify novel kinase inhibitors by high throughput screening have evolved rapidly in recent years. An important aspect is the availability of fluorescent probes that can be applied in a homogeneous, or mix-and-measure, assay format. Here, we illustrate the application of fluorescence read-out technologies for kinase targets in light of our own experiences in assay development and high-throughput screening. PMID- 12769675 TI - Competitive fluorescence polarization assays for the detection of phosphoinositide kinase and phosphatase activity. AB - We describe the development and implementation of competitive fluorescence polarization (FP) based assays for determining activity of phosphoinositide 3 kinase (PI 3-K) and the type-II SH2-domain-containing inositol 5-phosphatase (SHIP2). These assays are based on the interaction of specific phosphoinositide binding proteins with fluorophore-labeled phosphoinositide and inositol phosphate tracers. Enzyme reaction products are detected by their ability to compete with the fluorescent tracers for protein binding, leading to an increase in the amount of free tracer and a decrease in polarization (mP) values. A variety of fluorophore-labeled tracers were evaluated, and assay sensitivity and specificity for products of PI 3-K and SHIP2 activity was determined. Assay performance was evaluated using recombinant PI 3-Kalpha and SHIP2 with diC(8)-PI(4,5)P(2) and diC(8)-PI(3,4,5)P(3) as respective substrates. IC(50) values for previously characterized PI 3-K inhibitors were within expected ranges. These assays are homogeneous, sensitive, and rapid, and suitable for HTS applications, and will facilitate screening for novel inhibitors of phosphoinositide kinases and phosphatases in drug development. PMID- 12769676 TI - Strategies and solid-phase formats for the analysis of protein and peptide phosphorylation employing a novel fluorescent phosphorylation sensor dye. AB - Protein kinases represent one of the largest families of regulatory enzymes, with more than 2,000 of them being encoded for by the human genome. Many cellular processes are regulated by the reversible phosphorylation of proteins and upwards of 30% of the proteins comprising the eukaryotic proteome are likely to be phosphorylated at some point during their existence. In the past, analysis of global protein phosphorylation has been accomplished through radiolabelling of samples with inorganic (32P or [gamma-32)P] ATP. The approach is limited to specimens amenable to radiolabelling and poses certain safety and disposal problems. Alternatively, immunodetection with antibodies to the common phosphoamino acids may be employed, but the antibodies are relatively expensive and exhibit limited specificity and a certain degree of cross-reactivity. Pro-Q Diamond dye is a new fluorescent phosphosensor technology suitable for the detection of phosphoserine-, phosphothreonine- and phosphotyrosine-containing proteins directly in isoelectric focusing gels, SDS-polyacrylamide gels and two dimensional gels. Additionally, the technology is appropriate for the detection of phosphoproteins or phosphopeptides arrayed on protein chips or affixed to beads. Dye-stained proteins and peptides can be excited with a laser-based light source of 532 or 543 nm or with a xenon-arc lamp-based system equipped with appropriate band pass filters. Alternatively, ultraviolet light of about 302 nm may be employed, providing that sufficiently long exposure times are used to collect the fluorescence signal. Pro-Q Diamond dye emits maximally at approximately 580 nm. The fluorescence-based detection technology is easy to conduct, cost effective and allows rapid large-scale screening of protein and peptide phosphorylation in a variety of solid-phase assay formats. PMID- 12769677 TI - Development of fluorescence-based selective assays for serine/threonine and tyrosine phosphatases. AB - A number of aromatic substrates were evaluated for their ability to detect tyrosine phosphatase and serine/threonine phosphatase activity. Results demonstrated that the fluorinated coumarin DiFMUP is the most sensitive substrate for detecting LAR and PP-2A activity. Using this substrate, selective high throughput screening assays for serine/threonine and tyrosine phosphatases were developed. Specific inhibitor cocktails were added to each assay to limit the activity of other phosphatases. LAR, CD-45, and PTP-1B all rapidly hydrolyze DiFMUP in the tyrosine phosphatase assay. The activity of non-tyrosine phosphatases is less than 6% of the LAR activity. PP-1 and PP-2A are highly active in the serine/threonine phosphatase assay. Inhibition of LAR and PP-2A in these assays is demonstrated using known inhibitors. Both of these assays are sensitive, robust, kinetic assays that can be used to quantify enzyme activity. PMID- 12769679 TI - Measuring intracellular calcium fluxes in high throughput mode. AB - The measurement of intracellular calcium fluxes in real time is widely applied within the pharmaceutical industry to measure the activation of G-protein coupled receptors (GPCRhyp;s), either for pharmacological characterisation or to screen for new surrogate ligands. Initially restricted to G(q) coupled GPCRs, the introduction of promiscuous and chimeric G-proteins has further widened the application of these assays. The development of new calcium sensitive dyes and assays has provided sensitive, homogeneous assays which can be readily applied to high throughput screening (HTS). In this paper we describe the full automation of this assay type using a fluorometric imaging plate reader (FLIPR ) integrated into a Beckman/Sagian system to establish a simple robotic system that is well suited for the current medium throughput screening in this area of lead discovery. Using a recently completed HTS we discuss important determinants for FLIPR based screening, highlight some limitations of the current approach, and look at the requirements for future automated systems capable of keeping up with expanding compound files. PMID- 12769678 TI - Homogeneous fluorescence assay for cyclic AMP. AB - The objective of these studies was to develop a new homogeneous fluorescence assay for determining the concentrations of cAMP in biological samples. The assay is based on a novel general concept of using ligand-dependent sequence-specific DNA binding proteins as sensors for their respective ligands. CAP protein, a bacterial DNA binding protein whose DNA binding activity depends on cAMP, was used to develop the assay. In the presence of cAMP, DNA binding activity of CAP is greatly increased. Signaling of cAMP presence was achieved by detecting cAMP dependent formation of CAP-DNA complex using a recently developed fluorescence assay for DNA binding proteins (Heyduk, T., and Heyduk, E. Nature Biotechnology 20,171-176, 2002). Both 96-well and 384-well black microplate formats of the assay were developed and used to detect cAMP in low nanomolar concentrations. The assay involves mixing of the sample with the assay solution containing all necessary components for cAMP determination followed by fluorescence intensity readout; no washing or reagent addition steps are necessary. Excellent reproducibility of fluorescence signal change as a function of cAMP concentration was observed. Experiments with HEK 293 cells stimulated with forskolin were performed to demonstrate that the assay could be used for cAMP determination in cellular extracts. In summary, the obtained data fully validated the new homogenous assay for measuring cAMP based on cAMP-dependent DNA binding activity of CAP protein. It is expected that the development of assays for many other ligands of DNA binding proteins will be possible using the same overall assay design developed for cAMP. PMID- 12769680 TI - New Ca2+ fluoroionophores based on the BODIPY fluorophore. AB - Calcium (Ca(2+)) fluoroionophores are useful in cell-based functional assays of G protein coupled receptor (GPCR) activation or ion channel modulation. In this paper we describe new calcium probes that improve or overcome certain deficiencies in existing probes. These new fluoroionophores are based on acylation of amino-BAPTA [BAPTA = glycine, N,N'-(1,2-ethanediyl-bis(oxy-2,1 phenylene)) bis(N-(carboxymethyl))] with fluorescent BODIPY(R) propionates [BODIPY = 4,4-difluoro-5,7-dimethyl- 4-bora-3a,4a-diaza-s-indacene]. The resulting probes show high affinity to aqueous calcium solutions, and respond to calcium binding with significant fluorescence increases. The BODIPY fluorophores are uncharged and their fluorescence is pH-insensitive. The wide range of excitation/emission wavelength choices available within the BODIPY fluorophore series allows several different colors of new calcium indicators to be prepared. Cell permeable versions respond well with increasing fluorescence intensities in live cells after calcium influx. PMID- 12769681 TI - High throughput fluorescence-based assays for cyclic ADP-ribose, NAADP, and their metabolic enzymes. AB - Cyclic ADP-ribose (cADPR) and nicotinic acid adenine dinucleotide phosphate (NAADP) are two novel Ca(2+) messengers derived respectively from NAD and NADP. Since their discovery in sea urchin eggs, both have now been shown to serve messenger functions in a wide range of cells from plant to human. In this article, a series of fluorimetric assays for cADPR, NAADP and their metabolic enzymes is compiled. The enzyme assay makes use of an analog of NAD, nicotinamide guanine dinucleotide, which is non-fluorescent but is cyclized by the enzymes to a fluorescent analog of cADPR, cyclic GDP-ribose. Other NAD utilizing enzymes are not capable of catalyzing the cyclization and thus produce no interference. The fluorimetric assays for cADPR and NAADP make use of coupled-enzyme amplification and can readily detect nanomolar concentrations of either messenger. All the assays described can be performed in multi-well format, allowing ready automation and use in high throughput screening. An added advantage of these assays is that all the required reagents are commercially available, facilitating general adoption of the techniques by all those who are interested in the physiology and enzymology of the novel Ca(2+) signaling pathways mediated by cADPR and NAADP. PMID- 12769682 TI - Beta galactosidase enzyme fragment complementation as a novel technology for high throughput screening. AB - In this review, the applications of beta galactosidase complementation are described. alpha Complementation is a naturally occurring process in bacteria and in engineered cells, and can also occur in eukaryotic cells. Two forms of alpha complementation have been used in high throughput screening (HTS), in which interacting fragments complement with either low or high affinity. Low affinity complementation is used to monitor protein protein interactions, such as those occurring in homodimerization of the epidermal growth factor receptor (EGFR), and provides a robust screen for detection of EGFR inhibitors. High affinity complementation provides the basis for several HTS assays, in which analytes, such as cAMP or IP(3), are detected in crude cell lysates. A development of the latter approach is protein labeling, providing for measurement of cell protein expression and trafficking. Collectively, the use of beta galactosidase complementation provides a novel and flexible technology for highly sensitive, homogeneous HTS assay development. PMID- 12769683 TI - High throughput screening of G-protein coupled receptors via flow cytometry. AB - The molecular assemblies of signal transduction components, for example kinases and their target proteins or receptor-ligand complexes and intracellular signaling molecules, are critical for biological functions in cells. To better understand the interactions of these molecular assemblies and to screen for new pharmaceutics that could control and modulate these types of interactions, we have focused on developing high throughput approaches for the analysis of G protein coupled receptors via flow cytometry. Flow cytometry offers a number of advantages including real-time collection of multicomponent data, and together with improvements in sample handling, the high throughput sampling rate is up to 100 samples per minute. For our targets, assemblies of solubilized GPCRs, a screening platform of a dextran bead has proven to be flexible, allowing different surface chemistries on the beads. The bead can be either ligand-labeled or have epitope-linked proteins attached to the bead surface, enabling several molecular assemblies to be constructed and analyzed. A major improvement with this system is that for screening ligands for GPCRs the underlying mechanism of action for these compounds can be investigated and incorporated into the definition of a 'hit'. Our current screening system is capable of simultaneously distinguishing GPCR agonists and antagonists. PMID- 12769684 TI - Established and emerging fluorescence-based assays for G-protein function: heterotrimeric G-protein alpha subunits and regulator of G-protein signaling (RGS) proteins. AB - Heterotrimeric G-proteins are molecular switches that couple serpentine receptors to intracellular effector pathways and the regulation of cell physiology. Ligand bound receptors cause G-protein alpha subunits to bind guanosine 5'-triphosphate (GTP) and activate effector pathways. Signal termination is facilitated by the intrinsic GTPase activity of G-protein alpha subunits. Regulators of G-protein signaling (RGS) proteins accelerate the GTPase activity of the G-protein alpha subunit, and thus negatively regulate G-protein-mediated signal transduction. In vitro biochemical assays of heterotrimeric G-proteins commonly include measurements of nucleotide binding, GTPase activity, and interaction with RGS proteins. However, the conventional assays for most of these processes involve radiolabeled guanine nucleotide analogues and scintillation counting. In this article, we focus on fluorescence-based methodologies to study heterotrimeric G protein alpha subunit regulation in vitro. Furthermore, we consider the potential of such techniques in high-throughput screening and drug discovery. PMID- 12769685 TI - Established and emerging fluorescence-based assays for G-protein function: Ras superfamily GTPases. AB - Ras and Rho GTPases are signaling proteins that regulate a variety of physiological events and are intimately linked to the progression of cancer. Recently, a variety of fluorescence-based assays have been refined to monitor activation of these GTPases. This review summarizes current fluorescence-based techniques for studying Ras superfamily GTPases with an emphasis on practical examples and high-throughput applications. These techniques are not only useful for biochemical characterization of Ras superfamily members, but will also facilitate the discovery of small molecule therapeutics designed to inhibit signal transduction mediated by GTPases. PMID- 12769686 TI - The MYC oncogene as a cancer drug target. AB - The universal deregulation of c-myc gene expression in tumor cells suggests that this oncogene represents an attractive target for cancer therapeutic purposes. The same applies to the N-myc gene, which has a more restricted tissue specificity. Translocation (e.g., c-myc in Burkitt's lymphoma), or amplification (e.g., N-myc in neuroblastoma) of myc genes has been causally linked to tumor formation. Furthermore, the c-myc promoter integrates diverse mitogenic signalling cascades, which are constitutively activated in tumor cells, and translates them into expression of the c-MYC transcription factor, which promotes cell proliferation by regulating the expression of numerous target genes. Recent experimental data suggest, that even a brief inhibition of c-myc expression may be sufficient to permanently stop tumor growth and induce regression of tumors. Attempts to identify specific inhibitors of c-MYC/MAX dimerization have yielded promising results. In addition, downstream-target genes of c-MYC represent attractive targets for tumor therapy. Tumor cells expressing c-MYC at elevated levels are sensitized to treatment with DNA-damaging drugs. In mice and presumably also in human patients, the successful treatment of c-myc-induced tumors with conventional chemotherapy depends on the presence of functional p53. Therefore, restoration of this pathway, which is commonly lost in cancer cells, may enhance therapy of c-myc-induced tumors. These and other recent developments, which address the use of myc genes as therapeutic targets for cancer treatment, are discussed in this review. PMID- 12769687 TI - Gab1, SHP-2 and other novel regulators of Ras: targets for anticancer drug discovery? AB - Ras proteins function as molecular switches that cycle between an inactive GDP bound state, and an active GTP-bound form that triggers different signaling pathways. Because Ras can integrate both proliferative and anti-apoptotic stimuli, GTP-locked Ras mutants play a critical role in the development of human tumors. Moreover, wild-type Ras relays the transforming potential of a number of molecules involved in tumor development, including protein tyrosine kinases. Consequently, the molecular intermediates that control Ras activation are potential targets of anti-tumoral pharmacology. Besides the canonical Shc/Grb2/Sos module classically involved in Ras activation, novel effectors have recently been shown to participate in this pathway, including the multivalent Grb2-associated docking protein Gab1, the protein tyrosine phosphatase SHP-2, and the phosphoinositide 3-kinase. Recent genetic advances have shown that these proteins are critically involved in cell proliferation and survival, further suggesting that they could be interesting targets for selective tumor therapy. Here we review recent progress in our understanding of the role of Gab1 and its partners in Ras activation, and other survival/proliferation pathways. Implications for the pharmacological manipulation of this pathway in the treatment of cancer will also be discussed. PMID- 12769688 TI - Taxanes: microtubule and centrosome targets, and cell cycle dependent mechanisms of action. AB - Microtubules are highly dynamic cellular polymers made of alphabeta-tubulin and associated proteins. They play a key role during mitosis, participating in the exact organization and function of the spindle, and are critical for assuring the integrity of the segregated DNA. Therefore, they represent one of the more effective targets in current cancer therapy. Paclitaxel (Taxol) is the prototype of the taxane family of antitumor drugs, and it was the first natural product shown to stabilize microtubules. This unique mechanism of action is in contrast to other microtubule poisons, such as Vinca alkaloids, colchicine, and cryptophycines, which inhibit tubulin polymerization. Taxanes block cell cycle progression through centrosomal impairment, induction of abnormal spindles and suppression of spindle microtubule dynamics. Triggering of apoptosis by aberrant mitosis or by subsequent multinucleated G1-like state related to mitotic slippage, depends on cell type and drug schedule. The development of fluorescent derivatives of paclitaxel led us to locate spindle pole microtubules and centrosomes as main sub-cellular targets of cytotoxic taxoids in living cells. In this review we discuss these findings in the context of a cell cycle-dependent response to taxanes, based on the cellular targets, and the status of the implicated cell cycle checkpoints. We also review those events that can influence this response, like the different signal transduction pathways activated/inactivated in relation to Bcl-2 phosphorylation and induction of apoptosis, and the controversial role of the p53 status on cell sensitivity to paclitaxel. Finally, cell cycle-dependent resistance, an emerging concept in combination sequential chemotherapy, is discussed on the basis of the cell cycle dependent mechanisms of action of taxanes. PMID- 12769689 TI - Tumor angiogenesis: a potential target in cancer control by phytochemicals. AB - It is now well established that angiogenesis is an obligatory event for the growth and progression of solid tumors beyond the size limit (approximately 2 mm diameter) imposed by simple diffusion for the nutrient supply. Human tumors can remain dormant for years owing to a balance between cell proliferation and apoptosis. Several hypotheses have been articulated regarding the critical importance of tumor angiogenesis in the development and metastatic spread of tumors, and how preventive/therapeutic inhibition of angiogenesis might be exploited as a novel means of controlling cancer growth. Anti-angiogenic therapy is suggested as one of the most promising approaches to control cancer, as endothelial cells are generally non-transformed cells and are less prone to acquire drug resistance. Tumor vasculature could be an important prognostic marker, and an independent predictor of pathologic stages and malignant potential of cancer. This review is focused on recent developments and comprehensive mechanistic aspects of phytochemicals related to an interplay of angiogenic promoters and inhibitors, and associated signaling in both tumor as well as endothelial cells. Since, vascular endothelial cells constitute the first line exposure to the blood-borne agents, it is plausible that anti-angiogenic activity of phytochemicals could be associated with lowering the risk of cancer by preventing the growth and metastasis of tumor. PMID- 12769690 TI - Short-chain fatty acid inhibitors of histone deacetylases: promising anticancer therapeutics? AB - Cancer is a disease in which cellular growth regulatory networks are disrupted. Lesions in well-characterized oncogenes and tumor suppressors often contribute to the dysregulation, but recent work has also uncovered the fundamental importance of enzymes that modulate the acetylation status of chromatin to the initiation or progression of cancer. Histone acetyltransferases (HATs) and histone deacetylases (HDACs) are known to be involved in physiological cellular processes, such as transcription, cell cycle progression, gene silencing, differentiation, DNA replication, and genotoxic responses, but they are also increasingly being implicated in tumorigenesis. Butyrate is a short-chain fatty acid (SCFA) that acts as a HDAC inhibitor and is being clinically evaluated as an anti-neoplastic therapeutic, primarily because of its ability to impose cell cycle arrest, differentiation, and/or apoptosis in many tumor cell types, and its favorable safety profile in humans. Additionally, HDAC inhibitors could be used in combination with certain established antitumor therapeutics, such as those that target transcription, to augment clinical efficacy, and/or reduce toxicity. The molecular pathways of butyrate and related next-generation synthetic SCFAs in mediating these effects have not been fully elucidated, but HDAC inhibition is associated with regulation of critical cell cycle regulators, such as cyclin D, p21(CIP1/WAF1), and p27(KIP1). It is anticipated that a better understanding of this critical intersection between SCFAs, HDACs, and cell cycle control will lead to the design of novel treatment strategies for neoplasias. This review will summarize some of the recent research in these arenas of HDAC-directed cancer therapy and discuss the potential application of these agents in synergy with current chemotherapeutics. PMID- 12769691 TI - Human insulin genome sequence map, biochemical structure of insulin for recombinant DNA insulin. AB - Insulin is a essential molecule for type I diabetes that is marketed by very few companies. It is the first molecule, which was made by recombinant technology; but the commercialization process is very difficult. Knowledge about biochemical structure of insulin and human insulin genome sequence map is pivotal to large scale manufacturing of recombinant DNA Insulin. This paper reviews human insulin genome sequence map, the amino acid sequence of porcine insulin, crystal structure of porcine insulin, insulin monomer, aggregation surfaces of insulin, conformational variation in the insulin monomer, insulin X-ray structures for recombinant DNA technology in the synthesis of human insulin in Escherichia coli. PMID- 12769692 TI - Recent advances in the development of phytoestrogens and derivatives: an update of the promising perspectives in the prevention of postmenopausal diseases. AB - Phytoestrogens constitute a promising alternative in the treatment of diseases associated with menopause. Nevertheless, the lack of data concerning their pharmacology and their toxicology requires use precautions. After reminding the pharmacology of estrogen receptors, this review outlines the estrogenicity and the therapeutic potentialities of phytoestrogens according to their structure. PMID- 12769693 TI - Marine natural products as lead anti-HIV agents. AB - Current anti-HIV drugs have extreme side effects and resistance to these drugs develops rapidly. The marine environment holds an unprecedented number of unusual chemical structural classes with activity against HIV. We review the literature on anti-HIV activity of marine natural products and discuss the efficacy of different structural classes. PMID- 12769694 TI - Terpenoids and glycolipids from euphorbiaceae. AB - The family Euphorbiaceae is widely distributed throughout both hemispheres and ranges in morphological form from large desert succulents to trees and even small herbaceous types. Many species contain a milky juice which is more or less toxic, especially for cold-blooded animals, and can produce a dermatitis similar to that from poison ivy. Separation procedures and characterization of the less polar fractions of the plant extracts have been widely described in the literature for their content in diterpene derivatives. In the continuing research on biologically active compounds from Euphorbiaceae, a series of studies on the isolation and structure elucidation of glyceroglycolipids (GGLs) and glycosphingolipids (GSLs) have been carried out in order to develop the novel medicinal resources from natural Euphorbiaceae products. Glyceroglycolipids are major constituents of the chloroplast membrane in the plant kingdom. Recently, glycolipids were found to possess antitumor-promoting activity while glyceroglycolipids isolated from Euphorbiaceae have shown an interesting anti inflammatory activity in vivo. Glycosphingolipids are present at the outer layer of the lipid-bilayer in biological membranes and are thought to participate in antigen-antibody reactions and transmission of biologically informations. Sphingolipid breakdown products, sphingosine and lysosphingolipids, inhibit protein kinase C, a pivotal enzyme in cell regulation and signal transduction. Sphingolipids and lysosphingolipids affect significantly cellular responses and exhibit antitumor promoter activities in various mammalian cells. These molecules may function as endogenous modulators of cell function and possibly as second messengers. PMID- 12769695 TI - Crystallography of membrane proteins, major targets in drug design. AB - Protein crystallography has the potential to accelerate drug discovery greatly. High-resolution structures of membrane proteins of pharmaceutical interest open new perspectives in drug design. Recent structural data obtained for cyclooxygenases, monoamine oxidase, squalene cyclase, rhodopsine, porins, aquaporins, and ABC transporters are presented and briefly discussed. PMID- 12769696 TI - Microwave-assisted solid-phase synthesis (MASS): parallel and combinatorial chemical library synthesis. AB - The use of microwave technology in solid-phase organic synthesis has attracted much attention in recent years. The combination of solid support, either as a medium for chemical synthesis or as a carrier for organic reagents, with microwave heating offers several advantages over conventional techniques. Rapid and elevated heating of reaction mixtures can induce the completion of chemical transformations in minutes while several hours or days may be required for the same chemistry under conventional conditions. With decreased time of exposure to high temperatures and lessened thermal degradation, microwave accelerated chemistries often deliver products of higher purity when compared to conventional heating techniques. Several chemical syntheses on solid-phase employing microwave irradiation have been reported in the literature. The reagents, solvents, and equipment selected for microwave-mediated synthesis are important contributors to the success of the chemical transformation. Owing to the timesavings in performing chemical synthesis under microwave irradiation, the technique has become an emerging partner in solid-phase organic synthesis. PMID- 12769697 TI - The medicinal chemistry implications of the anticancer effects of aspirin and other NSAIDs. AB - The regular intake of aspirin and other non-steroidal anti-inflammatory drugs (NSAIDs) has been associated with decreased incidence of certain types of cancer particularly those with an inflammatory component. The protective effects of these drugs in colorectal cancer are particularly marked, with a 40-50% reduction in risk. Research in this area has focussed on understanding and optimising these cytoprotective effects. NSAIDs are believed to operate by inhibiting COX-2, an enzyme that appears to be involved in a number of cancer promoting processes. This hypothesis is consistent with the observation that the COX-2 selective inhibitors dramatically decrease tumour formation in human and animal studies. Surprisingly aspirin, which is selective for COX-1 over COX-2, and sulindac, which is an equipotent inhibitor of the COX isoenzymes, appear to have a similar anticancer profile to the COX-2 selective NSAIDs. A number of mechanisms have been proposed to explain the anomalous effects of aspirin. The first of these relates to the unique mode of action of aspirin, which acetylates the COX-2 enzyme and generates the cancer-suppressing 15R-hydroxyeicosatetraenoic acid at the site of a potential tumour. The alternative rationale relates to the metabolism of aspirin to salicylic acid, which has a cyclooxygenase independent anti-inflammatory mechanism, preventing the inflammatory response at the gene transcription level. A new generation of drugs could evolve from approaches to improving the therapeutic index of aspirin or by modifications to known therapies such as sulindac and celecoxib. PMID- 12769698 TI - O6-alkylguanine-DNA alkyltransferase inactivation in cancer chemotherapy. AB - The protein O(6)-alkylguanine-DNA alkyltransferase is the basis of an important process for repairing damage to cellular DNA, which renders cells resistant to drugs that alkylate at the O(6)-position of guanine residues. The development of various pseudosubstrates which inactivate this protein is reviewed, from a chemical standpoint. Study of the influence of pseudosubstrate molecular structure on their interaction with the active site cysteine has progressed together with direct investigation of protein structure. Combination therapy using a powerful inactivator with a suitable alkylating agent shows great clinical promise in the treatment of cancer, particularly when some degree of selectivity is possible. PMID- 12769699 TI - Review in quantitative structure activity relationships on lipoxygenase inhibitors. AB - This paper reviews and evaluates all the published QSAR treatments of LOX inhibitors. This reveals that in almost all cases, the clog P parameter plays an important part in the QSAR relationships. In some cases the steric factors (B(1), B(5) and L) as well as the overall molar refractivity (CMR) or the substitutents molar refractivity (MR) are important. Electronic effects except for the Hammet's constant alpha, are comparatively unimportant. The study shows that log P as calculated from the Clog P program is suitable for this form of QSAR study. Log Po of 2.77-3.76 was found to be ideal, for the biological response. PMID- 12769700 TI - Predicting passive transport in silico--history, hype, hope. AB - The development of computational tools for the prediction of passive transport is reviewed with particular reference to four diverse approaches: the rule-of-5, polar surface area, Volsurf and Abraham's General Solvation Equation. To illustrate the current state of the art, several examples of the application of in silico methods in drug design projects drawn from the recent medicinal chemistry literature are presented. In conclusion, the current challenges facing practitioners of this discipline are outlined and possible directions towards their resolution are suggested. PMID- 12769701 TI - Role of ADME characteristics in drug discovery and their in silico evaluation: in silico screening of chemicals for their metabolic stability. AB - Drug discovery is a long, arduous process broadly grouped into disease target identification, target validation, high-throughput identification of "hits" and "leads", lead optimization, and pre-clinical and clinical evaluation. Each area is a vast discipline in itself. However, all but the first two stages involve, to varying degrees, the characterization of absorption, distribution, metabolism, excretion, (ADME), and toxicity (T) of the molecules being pursued as potential drug candidates. Clinical failures of about 50% of the Investigational New Drug (IND) filings are attributed to their inadequate ADMET attributes. It is, therefore, no surprise that, in the current climate of social and regulatory pressure on healthcare costs, the pharmaceutical industry is searching for any means to minimize this attrition. Building mathematical models, called in silico screens, to reliably predict ADMET attributes solely from molecular structure is at the heart of this effort in reducing costs as well as development cycle times. This article reviews the emerging field of in silico evaluation of ADME characteristics. For different approaches that have been employed in this area, a critique of the scope and limitations of their descriptors, statistical methods, and reliability are presented. For instance, are geometry-based descriptors absolutely essential or is lower-level structure quantification equally good? What advantages, if any, do we have for methods like artificial neural networks over the least squares optimization methods with rigorous statistical diagnostics? Is any in silico screen worth application, let alone interpretation, if it is not adequately validated? Once deemed acceptable, what good is an in silico screen if it cannot be made available at the workbench of drug discovery teams distributed across the globe throughout multi-national pharmaceutical companies? These are not mere discussion points, rather this article embarks on the stepwise mechanics of developing a successful in silico screen. The process is exemplified by our efforts in developing one such screen for predicting metabolic stability of chemicals in a human S9 liver homogenate assay. A real life use of this in silico screen in a variety of discovery projects at GlaxoSmithKline is presented, highlighting successes and limitations of such applications. Finally, we project some capabilities of in silico ADME tools for greater impact and contribution to successful, efficient drug discovery. PMID- 12769702 TI - Prediction of drug metabolism: the case of cytochrome P450 2D6. AB - Cytochromes P450 (Cyt P450s) constitute the most important biotransformation enzymes involved in the biotransformation of drugs and other xenobiotics. Because drug metabolism by Cyt P450s plays such an important role in the disposition and in the pharmacological and toxicological effects of drugs, early consideration of ADME-properties is increasingly seen as essential for the discovery and the development of new drugs and drug candidates. The primary aim of this paper is to present various computational approaches used to rationalize and predict the activity and substrate selectivity of Cyt P450s, as well as the possibilities and limitations of these approaches, now and in the future. Attention is also paid to the experimental validation of these approaches by using high-throughput screening (HTS) of affinities to drug-drug interactions at the level of Cyt P450 isoenzymes. Since human Cyt P450 2D6 is one of the most important drug metabolizing enzymes and since in this regard much pioneering work has been done with this Cyt P450, Cyt P450 2D6 is chosen as a model for this discussion. Apart from early mechanism-based ab initio calculations on substrates of Cyt P450 2D6, pharmacophore modeling of ligands (i.e. both substrates and inhibitors) of Cyt P450 2D6 and protein homology modeling have been used successfully for the rationalisation and prediction of metabolite formation by this Cyt P450 isoenzyme. Significant protein structure-related species differences have been reported recently. It is concluded that not one computational approach is capable of rationalizing and reliably predicting metabolite formation by Cyt P450 2D6, but that it is rather the combination of the various complimentary approaches. It is moreover concluded, that experimental validation of the computational models and predictions is often still lacking. With the advent of novel, easily and well applicable in vitro based high throughput assays for ligand binding and turnover this limitation could be overcome soon, however. When effective links with other new and recent developments, such as bioinformatics, neural network computing, genomics and proteomics can be created, in silico rationalisation and prediction of drug metabolism by Cyt P450s is likely to become one of the key technologies in early drug discovery and development processes. PMID- 12769703 TI - Modeling biotransformation reactions by combined quantum mechanical/molecular mechanical approaches: from structure to activity. AB - An overview of the combined quantum mechanical/molecular mechanical (QM/MM) approach and its application to studies of biotransformation enzymes and drug metabolism is given. Theoretical methods to simulate enzymatic reactions have rapidly developed during the last decade. In particular, QM/MM methods provide detailed insights into enzyme catalyzed reactions, which can be extremely valuable in complementing experimental research. QM/MM methods allow the reacting groups in the active site of an enzyme to be studied at a quantum mechanical level, while the surrounding protein and solvent is included at a classical (and computationally less expensive) molecular mechanical level. Existing QM/MM implementations vary in the level of interaction between the QM and MM regions and in the way the partitioning into QM and MM regions is setup. Some general considerations concerning reaction modeling are discussed and a number of QM/MM studies related to drug metabolism are described. These studies illustrate that theoretical modeling of important metabolic reactions provides detailed insights into mechanisms of reaction and specific catalytic effects of enzyme residues as well as explaining variation in rates of conversion of different metabolites. Such information is essential in the development of methods to predict metabolism of drugs and to understand metabolic effects of genetic polymorphism in biotransformation enzymes. PMID- 12769704 TI - Progress in simulation modelling for pharmacokinetics. AB - Simulation models for the prediction of pharmacokinetics in humans and other mammalian species, which are based on the physiology and mechanistic models of absorption, distribution, metabolism and elimination are reviewed. The structure of such models is explained with reference to papers describing the mathematical details and alternative representations of organ flow and distribution. Approaches to the modelling of more complex tissues such as tumours and the liver are also reviewed as well as some specific transport processes such as biliary secretion and methods of ADME property estimation by experimental and in silico models. Specific approaches to the modelling of gastro-intestinal transit are explained as is the extension of the approach to simulating drug-drug interactions following co-administration of more than one drug. PMID- 12769705 TI - Molecular design and bioavailability. AB - A "snapshot" of current medicinal chemistry work on bioavailability is drawn from issues of J. Med. Chem. covering the time period between September 2001 and September 2002. An exhaustive compilation of reported absolute oral bioavailability (F) values for this period is included, covering 34 structural series and 107 distinct compounds, with data for multiple species in many cases. This is supplemented with a discussion of more qualitative oral bioavailability results, and with illustrative examples addressing clearance, prodrug design, and blood/brain barrier penetration problems. Papers discussing predictions pertaining to one or another aspect of bioavailability are also discussed, and some thoughts on future directions of work on in silico prediction in this area are presented. PMID- 12769706 TI - Designing safer drugs: (Q)SAR-based identification of mutagens and carcinogens. AB - Mutagenicity and carcinogenicity are chronic effects of primary concern for human health. A unifying approach to their mechanistic understanding is the recognition that many chemicals provoke both effects by electrophilic attack to the biological macromolecules, as such or after metabolism (genotoxic carcinogenicity). QSARs of individual classes of genotoxic carcinogens have contributed to the elucidation of the chemical determinants of this activity. Little work has been done on the epigenetic carcinogens, acting through non genotoxic, very specific mechanisms. However, the existing QSARs for individual chemical classes are too few to be of real usefulness in the screening of masses of candidate drugs. Models for predicting the carcinogenicity of "any type" of chemicals have been proposed: prospective prediction exercises pointed to the serious limitations of most of these approaches. The best alternative is provided by panels of human experts. The above prediction exercises considered samples of general chemicals, thus we specifically addressed in this paper the issue of pharmaceutical drugs. We applied our expert knowledge to a database of drugs whose carcinogenicity/noncarcinogenicity status was known. Whereas most of the noncarcinogens were correctly identified, our prediction of carcinogens was less successful than with the general chemicals. Several carcinogenic drugs did not show recognized structural alerts, and supposedly acted by epigenetic mechanisms. Whereas the contribution of human experts is highly valuable in this phase (e.g. priority setting), more work is necessary on: a) epigenetic carcinogens; b) efficient computerized models. PMID- 12769707 TI - Progress in toxinformatics: the challenge of predicting acute toxicity. AB - Historically, acute toxicity based on LC(50) and LD(50) tests has been analyzed using various quantitative structure-activity relationships (QSARs). The obtained QSAR equations cannot be related to the multiple health effects reflected in the experimental data of analyzed compounds. Therefore little predictive power for novel compounds can be achieved. New methods based on classification SAR (C-SAR) analysis offer new mechanistic knowledge that can be related to new health effects, resulting in better predictive power. To this end, a very careful interpretation of the obtained results is required, implying the use of the existing mechanistic information to the maximum possible extent. The current mini review aims at determining ways of automated extraction of new mechanistic knowledge from existing data, as well as ways of relating this knowledge to various health effects. A comparison of "statistical induction" and "knowledge based" approaches is provided. The existing and future developments in predictive acute toxicity are discussed. PMID- 12769708 TI - Use of synthetic isoprenoid analogues for understanding protein prenyltransferase mechanism and structure. AB - Protein prenylation involves the post-translational modification of specific protein-derived cysteine residues with farnesyl or geranylgeranyl groups through thioether linkages. Because a large number of proteins that participate in signal transduction processes require this modification, there has been intense interest in developing inhibitors of these enzymes and in clarifying the biological function of prenylation. Isoprenoid analogues have proven to be versatile tools for probing the mechanism and structure of prenyltransferases. Mechanistic probes have been created to investigate the stereochemical course and substituent effects in prenyltransferase catalyzed reactions. They have also been used to measure kinetic isotope effects and search for possible cationic intermediates. Photoaffinity labeling analogues containing either diazotrifluoropropionate or benzophenone units have been used to identify the location of isoprenoid binding sites in these enzymes. Biophysical probes incorporating fluorescent moieties or isotopic labels have been used to measure isoprenoid dissociation constants or prenyl group conformation when bound to the enzyme. Analogues containing noncognate alkene isomers or bulky substituents have also contributed to an understanding of isoprenoid recognition. Most recently, photoactive and isomeric isoprenylated analogues are providing insights into the function of protein prenylation. PMID- 12769709 TI - Inhibitors of farnesyltransferase and geranylgeranyltransferase-I for antitumor therapy: substrate-based design, conformational constraint and biological activity. AB - The development of farnesyltransferase inhibitors, a novel approach to non cytotoxic anticancer therapy, has been an active area of research over the past decade. Compounds that have advanced to clinical trials were evolved both from substrate-based design efforts and from compound library screening hits. This review focuses on the effort at Merck to evolve inhibitors from the protein substrate of farnesyltransferase, which resulted in the identification of a non peptide inhibitor for clinical evaluation. X-ray crystal structures of farnesyltransferase complexed with early peptidomimetic as well as later non peptide inhibitors have validated this design approach. NMR spectroscopic methods for studying enzyme-bound inhibitor structure, in conjunction with the use of conformational constraints, were critical components of subsequent efforts to provide potent inhibitors with varying levels of farnesyltransferase and geranylgeranyltransferase-I inhibitory specificity. Several of these compounds were important tools for investigating the use of prenyltransferase inhibitors to target Ki-Ras-mediated tumor growth. PMID- 12769710 TI - Farnesyl protein transferase inhibitor ZARNESTRA R115777 - history of a discovery. AB - R115777 (R)-6-amino[(4-chlorophenyl)(1-methyl-1H-imidazol-5-yl)methyl]-4-(3 chlorophenyl)-1-methyl-2(1H)-quinolinone is a potent and selective inhibitor of farnesyl protein transferase with significant antitumor effects in vivo subsequent to oral administration in mice. Taking its roots into Janssen's ketoconazole and retinoic acid catabolism programs, our interest into Ras prenylation process led us stepwise to identify the key structural features of R115777. Methodology, structure activity relationships, and pharmacology will be presented. R115777 is currently in phase III clinical evaluation. PMID- 12769711 TI - Sch-66336 (sarasar) and other benzocycloheptapyridyl farnesyl protein transferase inhibitors: discovery, biology and clinical observations. AB - Farnesyl Protein Transferase as a target for therapeutic intervention is currently under investigation in human clinical trials. Sch-66336 (sarasar), a benzocycloheptapyridyl Farnesyl Transferase Inhibitor (FTI), has been found to be effective in cellular proliferation assays and in in vivo oncology models both as a single agent and in combination with other anti-cancer agents. Clinically, early evidence is being generated that suggests efficacy in humans, particularly in patients with leukemia. Herein, we review the biology of FPT, the discovery of Sch-66336 and other benzocycloheptapyridyl FTIs, and the clinical evaluation of Sch-66336 for the treatment of leukemia and solid tumors. PMID- 12769712 TI - G-protein coupled receptor antagonists-1: protease activated receptor-1 (PAR-1) antagonists as novel cardiovascular therapeutic agents. AB - Inhibition of thrombin receptor (PAR-1) is a promising therapeutic approach for the treatment of various cardiovascular disorders such as unstable angina, acute myocardial infarction, stroke, and restenosis. Since a PAR-1 antagonist is specific for the cellulalr actions of thrombin, and does not interfere with fibrin generation, it is expected to have less bleeding liability than the currently available treatments. Several peptide and non-peptide PAR-1 antagonists with potent inhibition of platelet aggregation have been reported. Antithrombotic effect of a PAR-1 antibody has been demonstrated in a baboon thrombosis model and the antirestenosis property of a PAR-1 antagonist has been demonstrated in a rat model. PMID- 12769713 TI - The role of absorption, distribution, metabolism, excretion and toxicity in drug discovery. AB - Major reasons preventing many early candidates reaching market are the inappropriate ADME (absorption, distribution, metabolism and excretion) properties and drug-induced toxicity. From a commercial perspective, it is desirable that poorly behaved compounds are removed early in the discovery phase rather than during the more costly drug development phases. As a consequence, over the past decade, ADME and toxicity (ADMET) screening studies have been incorporated earlier in the drug discovery phase. The intent of this review is to introduce the desirable attributes of a new chemical entity (NCE) to the medicinal chemist from an ADMET perspective. Fundamental concepts, key tools, reagents and experimental approaches used by the drug metabolism scientist to aid a modern project team in predicting human pharmacokinetics and assessing the "drug-like" molecule are discussed. PMID- 12769714 TI - Structure-activity relationship studies: M2 and CCR5 receptor antagonists. AB - A wide range of neurotransmitters, polypeptides and inflammatory mediators transduce their signals into the interior of cell by specific interactions with cell-surface receptors that are coupled to G-protein. The most familiar G-protein coupled receptors are muscarinic receptors, adrenergic receptors, dopaminergic receptors and opioid receptors. A single polypeptide chain of 400-500 residues forms most of these receptors. There are seven hydrophobic regions in the receptor and they correspond to transmembrane alpha-helices, which are membrane spanning domains. This topology is highly conserved among various members of the family of G-protein coupled receptors. The amino-terminal extracellular domain contains potential N-linked glycosylation sites in most receptors. The carboxyl terminal is involved in the coupling to G-proteins and contains a cysteine site and phosphorylation site (Thr, Ser) and both are involved in receptor desensitization. In this section of the review we will discuss the development of potent, selective, low molecular weight antagonists of two G-protein coupled receptors (M(2) muscarinic receptor and CCR5 chemokine receptor) and their potential therapeutic utilities. The initial leads in both antagonist programs came from in house screening of our sample collections. As expected, most of the initial leads for both programs shared a similar pharmacophore and because of this showed strong affinity to many if not few a other G-protein coupled receptors. The initial significant challenge in both programs in terms of structure-activity studies was not only to optimize the structures for potency but also selectivity versus other subtype receptors. In the M(2) antagonist program the selectivity versus M(1) and other subtypes was a major challenge. Similarly in the CCR5 antagonist program the selectivity versus M(2) was a significant issue to overcome. In this review we will discuss in detail the structure activity relationships that resulted in potent and selective antagonists. PMID- 12769715 TI - Theoretical property predictions. AB - Methods for the prediction of octanol/water partition coefficient, aqueous solubility and acid/base dissociation constants are described and discussed. The advantages and limitations of the different approaches are described and an indication of problem areas discussed. Available prediction software is described and listed and attempts are made to assess the likely reliability of the predictions. The concept of "drug-likeness" is introduced and put into context and models for the prediction of ADME properties and toxicity are briefly described and assessed. Software for ADME/toxicity prediction is listed and the impact of these techniques on current drug design efforts is described. Web references are given for both commercial and public domain software which is available for property prediction from chemical structure. PMID- 12769716 TI - Computational analyses of high-throughput protein-protein interaction data. AB - Protein-protein interactions play important roles in nearly all events that take place in a cell. High-throughput experimental techniques enable the study of protein-protein interactions at the proteome scale through systematic identification of physical interactions among all proteins in an organism. High throughput protein-protein interaction data, with ever-increasing volume, are becoming the foundation for new biological discoveries. A great challenge to bioinformatics is to manage, analyze, and model these data. In this review, we describe several databases that store, query, and visualize protein-protein interaction data. Comparison between experimental techniques shows that each high throughput technique such as yeast two-hybrid assay or protein complex identification through mass spectrometry has its limitations in detecting certain types of interactions and they are complementary to each other. In silico methods using protein/DNA sequences, domain and structure information to predict protein protein interaction can expand the scope of experimental data and increase the confidence of certain protein-protein interaction pairs. Protein-protein interaction data correlate with other types of data, including protein function, subcellular location, and gene expression profile. Highly connected proteins are more likely to be essential based on the analyses of the global architecture of large-scale interaction network in yeast. Use of protein-protein interaction networks, preferably in conjunction with other types of data, allows assignment of cellular functions to novel proteins and derivation of new biological pathways. As demonstrated in our study on the yeast signal transduction pathway for amino acid transport, integration of high-throughput data with traditional biology resources can transform the protein-protein interaction data from noisy information into knowledge of cellular mechanisms. PMID- 12769717 TI - Nano-mechanical methods in biochemistry using atomic force microscopy. AB - The atomic force microscope has been extensively used not only to image nanometer sized biological samples but also to measure their mechanical properties by using the force curve mode of the instrument. When the analysis based on the Hertz model of indentation is applied to the approach part of the force curve, one obtains information on the stiffness of the sample in terms of Young's modulus. Mapping of local stiffness over a single living cell is possible by this method. The retraction part of the force curve provides information on the adhesive interaction between the sample and the AFM tip. It is possible to functionalize the AFM tip with specific ligands so that one can target the adhesive interaction to specific pairs of ligands and receptors. The presence of specific receptors on the living cell surface has been mapped by this method. The force to break the co operative 3D structure of globular proteins or to separate a double stranded DNA into single strands has been measured. Extension of the method for harvesting functional molecules from the cytosol or the cell surface for biochemical analysis has been reported. There is a need for the development of biochemical nano-analysis based on AFM technology. PMID- 12769718 TI - OB-fold: growing bigger with functional consistency. AB - It was predicted that the folding space for various protein sequences is restricted and a maximum of 1000 protein folds could be expected. Although, there were about 648 folds identified, general functional features of individual folds is not thoroughly studied. We selected OB-fold, which is supposed to be an oligonucleotide and oligosaccharide binding fold to study the general functional features. OB-fold is a small beta-barrel fold formed from 5 strands connected by modulating loops. We observed consistently 2 or 3 loops on the same face of barrel acting as clamps to bind to their ligands. Depending on the ligand, which could be a single or double stranded DNA/RNA or an oligosaccharide, and their conformational properties the loops change in length and sequence to accommodate various ligands. Different classes of OB-folded proteins were analyzed and found that the functional features are retained in spite of negligible sequence homology among various proteins studied. PMID- 12769719 TI - C-phycocyanin: a biliprotein with antioxidant, anti-inflammatory and neuroprotective effects. AB - Phycocyanin (Pc) is a phycobiliprotein that has been recently reported to exhibit a variety of pharmacological properties. In this regard, antioxidant, anti inflammatory, neuroprotective and hepatoprotective effects have been experimentally attributed to Pc. When it was evaluated as an antioxidant in vitro, it was able to scavenge alkoxyl, hydroxyl and peroxyl radicals and to react with peroxinitrite (ONOO(-);) and hypochlorous acid (HOCl). Pc also inhibits microsomal lipid peroxidation induced by Fe(+2)-ascorbic acid or the free radical initiator 2,2' azobis (2-amidinopropane) hydrochloride (AAPH). Furthermore, it reduces carbon tetrachloride (CCl(4))-induced lipid peroxidation in vivo. Pc has been evaluated in twelve experimental models of inflammation and exerted anti-inflammatory effects in a dose-dependent fashion in all of these. Thus, Pc reduced edema, histamine (Hi) release, myeloperoxidase (MPO) activity and the levels of prostaglandin (PGE(2)) and leukotriene (LTB(4)) in the inflamed tissues. These anti-inflammatory effects of Pc can be due to its scavenging properties toward oxygen reactive species (ROS) and its inhibitory effects on cyclooxygenase 2 (COX-2) activity and on Hi release from mast cells. Pc also reduced the levels of tumor necrosis factor (TNF-alpha) in the blood serum of mice treated with endotoxin and it showed neuroprotective effects in rat cerebellar granule cell cultures and in kainate-induced brain injury in rats. PMID- 12769720 TI - The biotin enzyme family: conserved structural motifs and domain rearrangements. AB - The biotin carboxylase family is comprised of a group of enzymes that utilize a covalently bound prosthetic group, biotin, as a cofactor. These enzymes, which include acetyl-CoA carboxylase, pyruvate carboxylase, propionyl-CoA carboxylase, methylcrotonyl-CoA carboxylase, geranoyl-CoA carboxylase, oxaloacetate decarboxylase, methylmalonyl-CoA decarboxylase, transcarboxylase and urea amidolyase, are found in diverse biosynthetic pathways in both pro-karyotes and eukaryotes. The reactions catalyzed by most members of this group of enzymes share two common features: (1) carboxylation of biotin, apparently via the formation of a carboxyphosphate intermediate, followed by (2) transcarboxylation of CO(2) from biotin to specific acceptor molecules to yield different products. Structural determinations by NMR and X-ray crystallography, complemented by mutagenesis studies, have identified some motifs that are structurally or catalytically important. Analysis of the amino acid sequences of a number of biotin carboxylases not only shows remarkable similarities within certain domains but also that there appears to have been domain rearrangements between groups of carboxylases. Acyl-coenzyme A derivatives, which bind either as substrates or as allosteric regulators of the biotin carboxylases, do not appear to share any of the CoA binding motifs that have been identified in other CoA-SH/acyl-CoA binding proteins. Further comparisons of biotin-dependent carboxylases with other groups of enzymes in the protein data bank reveal that this family of biotin enzymes has strong similarities in specific domains to a number of ATP-utilizing enzymes and to the lipoyl-containing enzymes. These structural homologies are so extensive as to be highly suggestive of evolutionary relationships between biotin carboxylases and these other enzymes. PMID- 12769721 TI - The bovine basic pancreatic trypsin inhibitor (Kunitz inhibitor): a milestone protein. AB - The pancreatic Kunitz inhibitor, also known as aprotinin, bovine basic pancreatic trypsin inhibitor (BPTI), and trypsin-kallikrein inhibitor, is one of the most extensively studied globular proteins. It has proved to be a particularly attractive and powerful tool for studying protein conformation as well as molecular bases of protein/protein interaction(s) and (macro)molecular recognition. BPTI has a relatively broad specificity, inhibiting trypsin- as well as chymotrypsin- and elastase-like serine (pro)enzymes endowed with very different primary specificity. BPTI reacts rapidly with serine proteases to form stable complexes, but the enzyme: inhibitor complex formation may involve several intermediates corresponding to discrete reaction steps. Moreover, BPTI inhibits the nitric oxide synthase type-I and -II action and impairs K+ transport by Ca2+ activated K+ channels. Clinically, the use of BPTI in selected surgical interventions, such as cardiopulmonary surgery and orthotopic liver transplantation, is advised, as it significantly reduces hemorrhagic complications and thus blood-transfusion requirements. Here, the structural, inhibition, and bio-medical aspects of BPTI are reported. PMID- 12769722 TI - Low-cost anti-HIV compounds: potential application for AIDS therapy in developing countries. AB - A considerable progress has been made in recent years in the field of drug development against HIV. However, the current cost of AIDS drugs is the main obstacle that prevents their use in developing countries, where 95% of HIV infected patients reside. The average yearly price of AIDS therapy and related health care of affected patients in the USA runs as high as $22,000 - an amount that corresponds to the combined income of as many as one hundred individuals in developing countries. Even in the USA, patients without medical insurance cannot afford the costly therapy. From the beginning it was clear that the most feasible and economic means of finding a solution is to identify anti-HIV drugs among already available and preferably over-the-counter pharmaceuticals, which have historically been used for unrelated clinical purposes. This review summarizes the development and discovery of affordable and potentially promising AIDS drugs. The anti-HIV activity of drugs and immunomodulating substances such as warfarin, cimetidine, levamisole, acetaminophen, gramicidin, and V-1 immunitor are described and discussed in relation to their clinical application. These compounds may be used in a cocktail drug combination. PMID- 12769723 TI - Concurrent use of antiretrovirals and anticonvulsants in human immunodeficiency virus (HIV) seropositive patients. AB - Seizure disorders may complicate HIV disease, either as a direct result of HIV or as a manifestation of a secondary opportunistic infection. Unless a reversible cause of seizure activity can be discerned, current treatment guidelines recommend the use of anticonvulsant drugs in these patients. The concurrent use of antiretrovirals and anticonvulsants is a poorly studied area. Controlled clinical trials examining drug-drug and drug-disease interactions in this area are scant, leaving clinicians a therapeutic dilemma in terms of drug selection. Most studies have been retrospective in nature. Generalized seizures appear to be most common and occur most frequently in patients with more severe disease as indicated by lower mean CD4(+) cell counts. In short follow-up periods, seizures appear to recur relatively frequently. Treatment of seizures in this population is hindered by a lack of clear data and numerous reports of drug-drug and drug disease interactions. In order to best provide evidence-based care, controlled clinical trials are needed to discern which anticonvulsants are best suited for use in this population. Trials should also examine appropriate dose adjustments that may be warranted when anticonvulsants and antiretrovirals agents are used concurrently. Unless an identifiable and reversible cause of seizures is identified in this patient population seizures should be treated with standard therapy and close follow-up and monitoring. Newer anticonvulsants (i.e., gabapentin, tiagabine) with fewer drug interactions may be better alternatives when compared to older anticonvulsant agents. Clinicians might avoid valproic acid given some conflicting reports regarding potential for increasing viral replication. PMID- 12769725 TI - Potential drug targets on the HIV-1 envelope glycoproteins, gp120 and gp41. AB - HIV-1 entry is an attractive target for anti-HIV-1 therapy. However, there are no entry inhibitors approved for the clinical treatment of HIV-1 infection. This is likely to be changed in the near future since promising HIV-1 entry inhibitors, such as T20 and some chemokine receptor antagonists, are in the pipeline to join the repertoire of anti-HIV-1 therapeutics. This review will focus on what might be potential targets on the key components of the viral entry machinery, gp120 and gp41. These two molecules are the viral proteins responsible for HIV-1 entry. Binding to CD4 induces a series of structural changes in gp120 and allows it to interact with chemokine receptors. The receptor binding eventually triggers conformational changes in gp41, which result in the formation of a fusion active molecule to attack the cell membrane. The structural and functional motifs that operate this delicate fusion machinery could become the Achilles' heel of the virus. PMID- 12769724 TI - Phospholipid analogs against HIV-1 infection and disease. AB - Phospholipid analogs are a new class of compounds with potent activity against HIV infection when used alone or conjugated with other therapeutic agents. When conjugated to the nucleoside analog AZT, the resulting phospholipid-AZT conjugate can double target the virus replication cycle by inhibiting the viral reverse transcriptase (by AZT) and inducing the production of defective virus particles that lack functional gp120 expression on the virus surface resulting in reduced capacity to bind to CD4+ cells and inhibition of infected cell-cell fusion (by phospholipid). Of great interest are data indicating that selected phospholipids are active against drug resistant variants, a current major problem in treating HIV/AIDS and controlling the epidemic occurring in various parts of the world. The purpose of this review is to provide current information on the design and synthesis of various types of phospholipids and phospholipid conjugates, in-vitro and in-vivo antiviral activity, tissue distribution, intracellular metabolism, and mechanism of action. The future development of this novel class of compounds offers an exciting approach for reducing the toxicity and enhancing the distribution of therapeutic drugs to the lymphatics and central nervous system and suppressing the emergence of drug resistant variants of HIV. PMID- 12769726 TI - Minidefensins: antimicrobial peptides with activity against HIV-1. AB - Over 80 different alpha-defensin or beta-defensin peptides are expressed by the leukocytes and epithelial cells of birds and mammals. Although their broad spectrum antimicrobial properties makes them candidates for therapeutic development, technical limitations related to their size (typically 30-45 residues) and complex structure have impeded such development. The minidefensins covered in this review are antimicrobial peptides with 16-18 residues, approximately half the number found in alpha-defensins. The theta-defensins are evolutionarily related toalpha- and beta-defensins, but other minidefensins probably arose independently. Like alpha- or beta-defensins, minidefensin molecules have a net positive charge and a largely beta-sheet structure that is stabilized by intramolecular disulfide bonds. Whereas alpha-defensins are found only in mammals and theta-defensins only in nonhuman primates, the other minidefensins come from widely divergent species, including horseshoe crabs, spiders, and pigs. Several alpha-defensins and minidefensins are effective inhibitors of HIV-1 infection in vitro, and recent evidence implicates alpha defensins in resistance to HIV-1 progression in vivo. This review compares defensins and minidefensins with respect to their activity against HIV-1. It pays special attention to retrocyclins - the ancestral theta-defensins of humans, and their extant counterparts in rhesus monkeys. In addition to describing critical elements of their structure and unusual mode of formation, we will venture some predictions about using theta-defensins as antiviral agents. PMID- 12769727 TI - Highly active antiretroviral therapy and cardiovascular complications in HIV infected patients. AB - Highly active antiretroviral therapy (HAART) has prolonged many patients' lives, but many cardiac sequelae of HIV are not affected by HAART and continue to develop even with treatment. In addition, HAART itself causes in a high proportion of patients a metabolic syndrome, characterized by lipodystrophy/ lipoatrophy, dyslipidemia and insulin resistance that may be associated with an increase in peripheral artery and coronary artery diseases. Careful cardiovascular evaluation in the course of HIV disease can identify cardiac complications early enough to treat. All HIV-infected patients who are either candidates to antiretroviral therapy or who are already under treatment should undergo an assessment that includes the evaluation of the cardiovascular risk with the available guidelines and the interactions between antiretrovirals and drugs commonly used to treat cardiovascular disease. PMID- 12769728 TI - Obesity and diabetes gene discovery approaches. AB - New treatments are currently required for the common metabolic diseases obesity and type 2 diabetes. The identification of physiological and biochemical factors that underlie the metabolic disturbances observed in obesity and type 2 diabetes is a key step in developing better therapeutic outcomes. The discovery of new genes and pathways involved in the pathogenesis of these diseases is critical to this process, however identification of genes that contribute to the risk of developing these diseases represents a significant challenge as obesity and type 2 diabetes are complex diseases with many genetic and environmental causes. A number of diverse approaches have been used to discover and validate potential new targets for obesity and diabetes. To date, DNA-based approaches using candidate gene and genome-wide linkage analysis have had limited success in identifying genomic regions or genes involved in the development of these diseases. Recent advances in the ability to evaluate linkage analysis data from large family pedigrees using variance components based linkage analysis show great promise in robustly identifying genomic regions associated with the development of obesity and diabetes. RNA-based technologies such as cDNA microarrays have identified many genes differentially expressed in tissues of healthy and diseased subjects. Using a combined approach, we are endeavouring to focus attention on differentially expressed genes located in chromosomal regions previously linked with obesity and/or diabetes. Using this strategy, we have identified Beacon as a potential new target for obesity and diabetes. PMID- 12769730 TI - Ghrelin as a potential anti-obesity target. AB - In order to develop an effective pharmacological treatment for obesity, an endogenous factor that promotes a positive energy balance by increasing appetite and decreasing fat oxidation could represent the drug target scientists have been looking for. The recently discovered gastric endocrine agent ghrelin, which appears to be the only potent hunger-inducing factor to naturally circulate in our blood stream, was discovered in 1999. Since then the acylated peptide hormone ghrelin has evolved from an endogenous growth hormone secretagogue to a regulator of energy balance to a pleiotropic hormone with multiple sources, numerous target tissues and most likely several physiological functions. Although neither the exact mechanism of action by which ghrelin increases food intake and adiposity is known, nor the putatively differential effects of brain-derived and stomach derived ghrelin on energy homeostasis have been determined, blocking or neutralizing ghrelin action still seems one of the more reasonable pharmacological approaches to reverse a chronically positive energy balance. However, based on growing experience with compounds targeting the neuroendocrine regulation of energy balance, it is quite possible that a ghrelin antagonist will either fail to cure obesity due to the existence of compensatory mechanisms or undesired effects might reveal the true biological function of ghrelin (e.g. cardiovascular mechanisms, anti-proliferative effects, reproduction). PMID- 12769729 TI - Central pre-proglucagon derived peptides: opportunities for treatment of obesity. AB - Modern societies have moved from famine to feast and obesity and its co morbidities now sweep the world as a global epidemic. Numerous scientific laboratories and pharmaceutical companies have taken the challenge and are now exploiting novel molecular targets for treatment of obesity. The pre-proglucagon system constitutes interesting candidates as potential targets for new anti obesity drugs. In the periphery, pre-proglucagon derived peptides, Glucagon-Like Peptide-1 (GLP-1), Glucagon-Like Peptide-2 (GLP-2) and oxyntomodulin (OXM) are involved in a wide variety of physiological functions, including glucose homeostasis, gastric emptying, intestinal growth, insulin secretion as well as the regulation of food intake. Peripheral administration of GLP-1 derivatives and analogues to both rodents and man have shown promising effects on food intake and body weight suggesting that such therapies constitute potential anti-obesity treatment. In the central nervous system, pre-proglucagon and hence GLP-1, GLP-2 and OXM are exclusively found in a small population of nerve cells in the nucleus of the solitary tract. These constitute a neural pathway linking the "viscero sensory" brainstem to hypothalamic nuclei involved in energy homeostasis. Intracerebroventricular administration of all of the three derived peptides robustly decrease food intake. It is evident that central GLP-1 agonism probably in combination with GLP-2 and/or OXM agonism constitute a potential pharmacological tool to reduce food intake and maybe also enhance energy expenditure. This and other aspects of the current state of the role of central pre-proglucagon in energy homeostasis are reviewed. PMID- 12769731 TI - Acylation stimulating protein and triacylglycerol synthesis: potential drug targets? AB - Triacylglycerol storage in adipose tissue is mediated by a host of transporters, enzymes and binding proteins. Additionally, several hormones (both autocrine and endocrine) are known to interact with cell surface receptors and modulate triacylglycerol synthesis (such as acylation stimulating protein, ASP). The many proteins involved contribute to the robustness of the system and, in most cases, deletion of a single gene is not deleterious and adipose tissue is preserved. On the other hand, this does not mean that gene disruption is not without effect, and in fact often results in a leaner, and presumably "healthier" mouse. These insights provide valuable indications for potential drug tools to delay and/or reverse obesity. In this review we examine the potential of ASP as a candidate target. ASP deficiency in mice decreases adipose tissue mass, increases insulin sensitivity and energy expenditure even in obese ob/ob mice, suggesting that partial interference of ASP action could be advantageous. ASP interacts with a specific cell surface receptor present in adipose tissue and certain structural components, such as the tightly folded core region, are implicated in activity. We propose that interference of the ASP-receptor interaction using an antagonist offers future prospect for an anti-obesity target. PMID- 12769732 TI - The insulin-sensitizing role of the fat derived hormone adiponectin. AB - Adiponectin is an insulin-sensitizing hormone whose blood concentration is reduced in obesity and type 2 diabetes. Administration of recombinant adiponectin in rodents increases glucose uptake and increases fat oxidation in muscle, reduces fatty acid uptake and hepatic glucose production in liver, and improves whole body insulin resistance. The exact receptor and signaling systems are unknown, however, recent studies suggest adiponectin activates AMPK, a putative master metabolic regulator. Thus, excitement surrounds the potential for adiponectin, or a homologue of adiponectin, as pharamacotherapy agents for patients suffering from the metabolic syndrome and more particularly for individuals with insulin resistance and type 2 diabetes. PMID- 12769733 TI - Antimicrobial peptides from food proteins. AB - Antimicrobial peptides are present in men, animals and plants and represent an important component of the innate immunity. Nevertheless they can also be generated through proteolytical digestion of food proteins. Thus, food proteins can be regarded not only for their nutritive value but also as a possible resource to increase the natural defence of the organism against invading pathogens. Consequently food proteins can be considered as component of nutritional immunity. Antimicrobial peptides generated from food proteins present the great advantage to be derived from harmless substances, therefore one can expect their safety for use in medicine and in food industry. Many biologically active peptides have been produced from food proteins, in particularly from milk proteins. The possibility that proteins can be tailored and their fragments modelled to achieve a particular function is recently giving rise to increased interest. This strategy has had particular success with food proteins like lactoferrin and lysozyme. Both bactericidal domains of these proteins have been extensively investigated. A number of short peptides with high bactericidal activity have been developed from the bactericidal domain of lysozyme through the strategy "tailoring and modelling". Ovotransferrin, alpha--lactalbumin and beta lactoglobulin are further examples of food proteins which are a source of antimicrobial peptides. The observation that antimicrobial peptides can be generated through proteolytical digestion of parent proteins, which usually have another physiological function in the organism, led us to consider these latter as multifunctional molecules. This raises the question, whether multifunctionality is an intrinsic property of many proteins or limited to a few. PMID- 12769734 TI - Biodefense properties of milk: the role of antimicrobial proteins and peptides. AB - Mammary fluids, colostrum and milk, deliver nature's first host defense systems upon birth, and these essential liquids are critical for survival of the neonate. The identification and characterization of anti-infectious proteins were among the early scientific discoveries and this group of proteins has long been recognized for promoting health benefits in both newborns and adults. Among the more widely studied are the immunoglobulins, lactoperoxidase, lysozyme, and lactoferrin. Recently, it was shown that alpha--lactalbumin may also function in a protective capacity dependent upon its folding state. Some of these, especially lactoferrin, also display an immunomodulatory role in which case a totally separate cascade of host defense responses is initiated. It was noted that the mechanism of action for this cluster of sentry proteins does vary; thus, this protective strategy provides for a broad range of responsive reactions to infection. Presently, there is a major focus on the discovery of novel peptides that can be generated from existing milk proteins via proteolytic reactions. To date, this substrate list includes alpha--lactalbumin, beta-lactoglobulin, all casein fractions, and lactoferrin. Again, the immunoregulatory effects prompted as a result of the appearance of these peptides are currently being defined. Herein, we review the principal biological properties associated with each of these contributing milk components with a special emphasis on the role of biodefensive milk peptides. We envision future contributions emerging from this research field as an opportunity to develop effective new therapies to be used in treating infectious diseases and promoting health benefits in vivo. PMID- 12769735 TI - Antibacterial and antiviral effects of milk proteins and derivatives thereof. AB - Milk forms a rich source of biologically interesting components. In particular, its protein fraction is known to encompass many kinds of biological functions. In this review we focus on antibacterial and antiviral properties of milk proteins and milk protein derivatives. The latter include chemically modified proteins and enzymatically induced peptides. If such peptides are released by enzymes present within the digestive tract (e.g. trypsin or pepsin), it is likely that they play a role in the health defense system. This is especially the case when the active fragments can survive the intestinal conditions long enough to arrive at the right place to exert their beneficial function. In the first part of this paper attention is paid to the antibacterial proteins lactoferrin, lactoperoxidase, and lysozyme. Furthermore, antibacterial peptides originating from caseins and whey proteins are described. The second part reports on studies of antiviral effects of milk proteins and derivatives thereof. Special focus is directed to the antiviral action towards the human immunodeficiency virus (HIV) and the human cytomegalovirus (HCMV). Unmodified milk proteins are generally not active against these viruses. An exception is lactoferrin, which shows significant antiviral activity against both HIV and HCMV. Several other milk proteins tested showed strong antiviral effects only after chemical modification, i.e. by making them polyanionic (for anti-HIV activity) or polycationic (for anti-HCMV activity). In a number of cases, conclusions are drawn concerning possible relationships between antibacterial/antiviral activity and molecular structure of the components described. PMID- 12769736 TI - Lactoferricin derived from milk protein lactoferrin. AB - Lactoferricin (LFcin) was initially identified as an antimicrobial peptide derived by pepsin digestion of lactoferrin (LF), a multifunctional innate-defense protein in milk. Various synthetic analogs of LFcin have also been reported. LFcin inhibits a diverse range of microorganisms such as gram-negative bacteria, gram-positive bacteria, yeast, filamentous fungi, and parasitic protozoa, including some antibiotic-resistant pathogens. LFcin kills target organisms by membrane perturbation and acts synergistically with some antimicrobial agents. LFcin exhibits numerous biological activities in common with those of LF. Whereas LFcin suppresses the activation of innate immunity by microbial components such as lipopolysaccharide (LPS) and CpG DNA, the peptide itself activates immunity. Administration of LFcin analogs has been shown to protect the host via direct antimicrobial activity and immunostimulatory effects in several infection models of mice. Here we present a comprehensive review of investigations of LFcin and related peptides. PMID- 12769737 TI - Biofunctional peptides from milk proteins: mineral binding and cytomodulatory effects. AB - The protein fraction of milk contains many valuable components and biologically active substances. Moreover, milk proteins are precursors of many different biologically active peptides which are inactive within the sequence of the precursor protein but can be released by enzymatic proteolysis. Many milk protein derived peptides, such as caseinophosphopeptides, reveal multi-functional bioactivities. Caseinophosphopeptides can form soluble organophosphate salts and may function as carriers for different minerals, especially calcium. Furthermore, they have been shown to exert cytomodulatory effects. Cytomodulatory peptides inhibit cancer cell growth or they stimulate the activity of immunocompetent cells and neonatal intestinal cells, respectively. Several bioactive peptides derived from milk proteins are potential modulators of various regulatory processes in the body and thus may exert beneficial physiological effects. Caseinophosphopeptides are already produced on an industrial-scale and as a consequence these peptides have been considered for application as ingredients in both 'functional foods' and pharmaceutical preparations. Although the physiological significance as exogenous regulatory substances is not yet fully understood, both mineral binding and cytomodulatory peptides derived from bovine milk proteins are claimed to be health enhancing components that can be used to reduce the risk of disease or to enhance a certain physiological function. PMID- 12769738 TI - Food-derived bioactive peptides--opportunities for designing future foods. AB - Dietary proteins are known to carry a wide range of nutritional, functional and biological properties. Nutritionally, the proteins are a source of energy and amino acids, which are essential for growth and maintenance. Functionally, the proteins contribute to the physicochemical and sensory properties of various protein-rich foods. Furthermore, many dietary proteins possess specific biological properties which make these components potential ingredients of functional or health-promoting foods. Many of these properties are attributed to physiologically active peptides encrypted in protein molecules. Particularly rich sources of such peptides are milk and egg, but they are also found in meat of various kinds as well as many plants. These peptides are inactive within the sequence of parent protein and can be released during gastrointestinal digestion or food processing. Depending on the amino acid sequence, these peptides may exert a number of different activities in vivo, affecting, e.g., the cardiovascular, endocrine, immune and nervous systems in addition to nutrient utilization. There is increasing commercial interest in the production of bioactive peptides from various sources. Industrial-scale production of such peptides is, however, hampered by the lack of suitable technologies. Bioactive peptides can also be produced from milk proteins through fermentation of milk, by starters employed in the manufacture of fermented milks or cheese. In particular, antihypertensive peptides have been identified in fermented milk, whey and ripened cheese. A few of these peptides have been commercialised in the form of fermented milks. There is a need to develop technologies which retain or even enhance the activity of bioactive peptides in food systems. Also, it is essential to study the optimum utilization of such peptides during passage through the gastrointestinal tract. PMID- 12769739 TI - Bioactive proteins and peptides from food sources. Applications of bioprocesses used in isolation and recovery. AB - There are many examples of biologically active food proteins, with physiological significance beyond the pure nutritional requirements that concern available nitrogen for normal growth and maintenance. Moreover, there are many physiologically active peptides, derived by protease activity from various food protein sources; however, relationships between structural properties and functional activities have not been completely elucidated. Many bioactive peptides have in common structural properties that include a relatively short peptide residue length (e.g. 2-9 amino acids), possessing hydrophobic amino acid residues in addition to proline, lysine or arginine groups. Bioactive peptides are also resistant to the action of digestion peptidases. Antihypertensive peptides, known as Angiotensin I converting enzyme (ACE) inhibitors have been derived from milk, corn and fish protein sources. Peptides with opioid activities are derived from wheat gluten or casein, following digestion with pepsin. Exorphins, or opioid peptides derived from food proteins such as wheat and milk (e.g. exogenous sources) have similar structure to endogenous opioid peptides, with a tyrosine residue located at the amino terminal or bioactive site. Immunomodulatory peptides derived from tryptic hydrolysates of rice and soybean proteins act to stimulate superoxide anions (reactive oxygen species-ROS), which triggers non-specific immune defense systems. Antioxidant properties that prevent peroxidation of essential fatty acids have also been shown for peptides derived from milk proteins. The addition of a Leu or Pro residue to the N-terminus of a His-His, dipeptide will enhance antioxidant activity and facilitate further synergy with non-peptide antioxidants (e.g. BHT). We also show herein, that the tryptic digests of casein yielding caseinophosphopeptides exhibits both hydrophilic and lipophilic antioxidant activity due to both metal ion sequestering and quenching of ROS. The separation and purification of bioactive peptides which will involve development of automated and continuous systems is an important field for Food chemists. Much effort has been given to develop selective column chromatography methods that can replace batch methods of salting out, or using solvent extraction to isolate and purify bioactive peptides. Advances here will enable recovery of bioactive peptides with minimal destruction thus enabling utilization by returning these active peptides to functional food or specific nutraceutical applications. PMID- 12769740 TI - Delta opioid peptides derived from plant proteins. AB - Opioid peptides showing selectivity for delta receptor have been isolated from enzymatic digests of plant proteins. Five peptides were derived from wheat gluten, and named gluten exorphins A5, A4, B5, B4 and C. Two opioid peptides were also released from spinach ribulose-bisphosphate-carboxylase/oxygenase (Rubisco), and named rubiscolins-5 and -6. Among them, gluten exorphin 5A (Gly-Tyr-Tyr-Pro Thr) and rubiscolin 6 (Tyr-Pro-Leu-Asp-Leu-Phe) improved learning performance in step-through type passive avoidance test after post-training oral administration in mice at doses of 300 mg/kg and 100 mg/kg, respectively, which are smaller than those required for antinociceptive activity. PMID- 12769741 TI - Opioid receptor ligands derived from food proteins. AB - During the last two decades a variety of food protein fragments has been demonstrated to elicit biological effects in various in vitro or in vivo test systems. A considerable part of these bioactive peptides are opioid receptor ligands, which may be regarded as exogenous supplements to the endogenous opioidergic systems of the human organism. Most of these food-derived opioid receptor ligands are fragments of the milk proteins alpha-, beta- or kappa casein, alpha-lactalbumin, beta-lactoglobulin or lactotransferrin; however, also wheat gluten, rice albumin, bovine serum albumin or hemoglobin, i.e. possible constituents of meat, and even a protein from spinach could be demonstrated to contain fragments behaving like opioid receptor ligands. Practically all of these compounds display opioid agonist activity; only very few of them behave like opioid antagonists. Bioactive food protein derivatives have been termed " food hormones", which implies that these compounds display their bioactivities when released from food constituents, i.e. from their precursor molecules due to the action of gastrointestinal enzymes. The critical point in case of food protein derived opioid receptor ligands is that only a minority of their bioactive effects demonstrated as yet has been observed upon oral or intragastric administration of these peptides or their precursor proteins and that most of these studies have been performed in animals. Thus, in terms of "evidence-based dietary supplementation" more studies are needed to prove effects of food protein derived opioid receptor ligands or their precursors after oral administration in humans and, moreover, to prove a benefit for the consumer's organism. PMID- 12769743 TI - Treatment of obesity: should we target the individual or society? AB - Obesity is a heritable disease that affects millions of people, is disproportionately prevalent in some ethnic groups, and has serious health consequences. Molecular mechanisms causing excessive adiposity are being discovered at an unprecedented rate in animal models. The same cannot be said for humans and, in fact, the etiology of obesity in the majority of people remains unknown. Furthermore, we are far from fully understanding how an obesogenic environment increases the severity of the disease in people who are genetically susceptible to weight gain. Due to these uncertainties, it is perhaps not surprising that current antiobesity treatments are moderately effective at best. This manuscript provides a brief review of current and future strategies for the treatment of obesity and how they relate to our current knowledge of its pathophysiology. It is concluded that there are reasons to be moderately optimistic that effective pharmacological means to palliate obesity will eventually be identified. However, reversing the current pandemic will require a greater understanding not only of the molecular and physiological underpinnings, but also social and political causes of this disease. PMID- 12769742 TI - Biogenic peptides and their potential use. AB - This paper reviews bioactive peptides, biogenic peptides, opioid peptides, immunostimulating peptides, mineral soluble peptides, antihypertensive peptides and antimicrobial peptides originating from food materials and enzymatic hydrolysis of proteins. Antihypertensive peptides are extensively reviewed and have been divided into angiotensin I-converting enzyme inhibitory peptides and others. These peptides are produced in the enzymatic hydrolysate of treated food materials such as milk, animal and fish meat, maize, wheat, soybeans and egg, and also from microbe-fermented products. Peptides with strong antihypertensive effects on spontaneously hypertensive rats are discussed and are divided into high and low angiotensin I-converting enzyme inhibitory activities. In addition, new topics from our studies on antihypertensive peptides are introduced. Efficacies of these peptides in clinical studies and differences with medicinal substances are summarized. Recent studies in this area shown the possibility of using biogenic peptides for improvements in treatment or prevention of hypertension. PMID- 12769744 TI - Neuropeptide Y (NPY) family of hormones: progress in the development of receptor selective agonists and antagonists. AB - Neuropeptide Y (NPY), the most abundant peptide present in the mammalian brain, exhibits a wide spectrum of central and peripheral activities mediated by at least six G-protein coupled receptors. The latter observation, and the implication of NPY in the pathophysiology of feeding, seizures, diabetes, intestinal dysfunction, cardiovascular diseases and respiratory disorders, have led to vigorous efforts to dissociate various effects of NPY and develop receptor selective ligands required for fundamental investigations, and possible clinical utility. These efforts have made significant advancement in the development of antagonists, especially for Y(1) and Y(5) receptors mediating NPY effects on feeding and/or thermogenesis. However, only a limited progress has been made in the case of Y(2) ligands, and none in the case of Y(4) ligands. Moreover, most of the nonpeptidic ligands developed to date have little use clinically because of their solubility and toxicity problems and their limited passage through blood brain barrier. Furthermore, no progress has been made in developing lower molecular weight agonists, which may also have clinical potential in treating seizures, intestinal dysfunction, respiratory disorders, cachexia and anorexia. Thus, despite significant advances, NPY research is expected to attract scientists for years to come in the pursuit to develop clinically useful ligands. The recent advances in the peptide drug delivery techniques have given added impetus for these efforts. This article reviews the usefulness of widely used ligands as well as those developed more recently. PMID- 12769745 TI - Glucose analog inhibitors of glycogen phosphorylases as potential antidiabetic agents: recent developments. AB - Diabetes is among the largest contributors to global mortality through its long term complications. The worldwide epidemic of type 2 diabetes has been stimulating the quest for new concepts and targets for the treatment of this incurable disease. A new target is glycogen phosphorylase (GP), the main regulatory enzyme in the liver responsible for the control of blood glucose levels. One of several approaches to influence the action of GP is the use of glucose derivatives as active site inhibitors. This field of research commenced 10-15 years ago and, due to joint efforts in computer aided molecular design, organic synthesis, protein crystallography, and biological assays, resulted in glucopyranosylidene-spiro-hydantoin 16 (K(i) = 3-4 micro M) as the most efficient glucose analog inhibitor of GP of that time. The present paper surveys the recent developments of this field achieved mainly in the last five years: the synthesis and evaluation of glucopyranosylidene-spiro-thiohydantoin 18 (K(i) = 5 micro M) which has proven equipotent with 16, and is available in gram amounts; furanosylidene- and xylopyranosylidene-spiro-(thio)hydantoins whose ineffectiveness (K(i) > 10 mM) confirmed the high specificity of the catalytic site of GP towards the D-glucopyranosyl unit; "open" hydantoins like methyl N-(1 carboxamido-D-glucopyranosyl)carbamate 37 (K(i) = 16 micro M) and N-acyl-N'-(beta D-glucopyranosyl)ureas among them the to date best glucose analog inhibitor N-(2 naphthoyl)-N'-(beta-D-glucopyranosyl)urea (35, K(i) = 0.4 micro M) which can also bind to the so-called new allosteric site of GP; C-(beta-D glucopyranosyl)heterocycles (tetrazole, 1,3,4-oxadiazoles, benzimidazole (K(i) = 11 micro M), and benzothiazole). Iminosugars like isofagomine (45, IC(50) = 0.7 micro M), noeuromycin (53, IC(50) = 4 micro M), and azafagomine (54, IC(50) = 13.5 micro M) also bind strongly to the active site of GP, however, substitution on the nitrogens makes the binding weaker. The natural product five-membered iminosugar DAB (56) exhibited IC(50) approximately 0.4-0.5 micro M. Azoloperhydropyridines which can be regarded iminosugar-annelated heterocycles show moderate inhibition of GP: nojiritetrazole 12 (K(i) = 53 micro M) is the best inhibitor and fewer nitrogens in the five-membered ring weakens the binding. Physiological investigations have been carried out with N-acetyl-beta-D glucopyranosylamine 6, spiro-thiohydantoin 18, isofagomine 45, and DAB 56 to underline the potential use of these compounds in the treatment of type 2 diabetes. Computational methods suggest to synthesize further anomerically bifunctional glucose derivatives which may be good inhibitors of GP. PMID- 12769746 TI - Nonpeptidic chymase inhibitors: design and structure-activity relationships of pyrimidinone derivatives based on the predicted binding mode of a peptidic inhibitor. AB - While the biological reaction of chymase have been often studied for ten years, the pathophysiological role of chymase has not been fully elucidate due to a lack of effective inhibitors featuring potent inhibitory activity, specificity, and metabolic stability. Recently the discovery of a structurally varied range of novel nonpeptidic inhibitors presents new opportunities to explore the role of chymase under both physiological and pathophysiological conditions and to develop therapeutic agents for chymase-induced diseases. In this article the structure and the inhibitory mechanism of nonpeptidic chymase inhibitors are discussed, with special emphasis on design and structure-activity relationships of pyrimidinone derivative where inhibitory activity, protease selectivity, and pharmacokinetic profile are clarified. PMID- 12769747 TI - CCR1 chemokine receptor antagonist. AB - The selective accumulation and activation of leukocytes in inflamed tissues contributes to the pathogenesis of inflammatory and autoimmune diseases such as infection, rheumatoid arthritis, allergic asthma, atopic dermatitis, and multiple sclerosis. A substantial body of reports suggests that chemokines and their receptors, which belong to a family of seven transmembrane G-protein coupled receptors (GPCR), may be involved in the selective accumulation and activation of leukocytes in inflamed tissues, and in the pathogenesis of inflammatory and autoimmune diseases. One such receptor is CCR1 which is a receptor for CC chemokines, such as CCL5 (RANTES) and CCL3 (MIP-1alpha). The involvement of CCR1 in immunological diseases now is documented in several preclinical studies with CCR1 deficient mice, anti-CCR1 antibodies and CCR1 antagonists, suggesting that CCR1 may be an attractive therapeutic target for a variety of diseases. Publications and patents describing CCR1 antagonists and their pharmacological effects have recently been disclosed. This review highlights the biology and pathophysiology of CCR1, and some of its currently reported antagonists. Additionally, our approach to CCR1 drug discovery is summarized. PMID- 12769748 TI - Design of beta-turn based therapeutic agents. AB - Peptides and proteins are essential to many biological processes. The interaction between the peptide ligands and their receptor targets commonly involves beta turn structures. Yet poor bioavailability and unfavorable pharmacokinetics significantly compromise the use of peptides as drugs. Thus, there has been a great deal of interest in the design of peptidomimetics (modified peptides) as therapeutic agents by mimicking beta-turn structures. This review highlights the importance of beta-turn in the design of various peptidomimetics for many diseases. This review also outlines several beta-turn mimicking strategies and its application in the design of potent peptide analogues. beta-turn mimetics often tend to be more rigid in positioning the critically important amino acid residues and thus optimize the surface conformation for productive interaction with the receptors. PMID- 12769749 TI - Anti-cytokines and cytokines in the treatment of rheumatoid arthritis. AB - The treatment approach to rheumatoid arthritis has undergone a major evolutionary change in recent years in part as a consequence of growing appreciation of the severity of this condition and in part due to very considerable progress in understanding the important role of cytokines in the immunopathogenesis of this disease. The major focus of this review is on the rationale for targeting TNFalpha and IL-1 in rheumatoid arthritis and the results of clinical studies designed to assess the validity of this therapeutic approach. Recently published studies confirm that the long term use of a several biological agents targeting TNFalpha give rise to sustained improvements in symptoms and signs of rheumatoid disease and, furthermore, that TNFalpha blockade protects joints from structural damage. Although these drugs are well tolerated and have a good overall safety profile, pitfalls to the use of anti-TNFalpha agents apparent with increasing clinical experience include rare cases of tuberculosis. The mechanism of action of anti-TNFalpha therapy is discussed. Clinical trials of interleukin-1 receptor antagonist show relatively modest anti-inflammatory efficacy and an effect on X ray indicative of retardation of joint damage. Other pro-inflammatory cytokines representing potential therapeutic targets include interferon-beta, interferon alpha, IL-6, IL-15, IL-17 and IL-18. I will consider preliminary data, where available, arising from clinical trials designed to inhibit the activity of such molecules. In this review I will also discuss the rationale and preliminary data for other potential therapeutic strategies designed to augment the activity of anti-inflammatory cytokines such as IL-4, IL-10, and IL-11 in rheumatoid disease. PMID- 12769750 TI - Cytokine and anti-cytokine therapies for inflammatory bowel disease. AB - Although the pathogenesis of inflammatory bowel disease (IBD) remains elusive, it appears that there is chronic activation of the immune and inflammatory cascade in genetically susceptible individuals. Current disease management guidelines have therefore focused on the use of anti-inflammatory agents, aminosalicylates and corticosteroids. These conventional therapies continue to be a first choice in the management of IBD. Immunomodulators, such as azathioprine, 6 mercaptopurine, methotrexate or cyclosporin, are demonstrating increasing importance against steroid-resistant and steroid-dependent patients. However, some patients are still refractory to these therapies. Recent advances in the understanding of the pathophysiological conditions of IBD have provided new immune system modulators as therapeutic tools. Other immunosuppressive agents including FK506 and thalidomide have expanded the choice of medical therapies available for certain subgroups of patients. Furthermore, biological therapies have begun to assume a prominent role. Studies with chimeric monoclonal anti-TNF alpha antibody treatment have been reported with dramatic successes. However, observations in larger numbers of treated patients are needed to explicate fully the safety of or risks posed by this agent such as developing lymphoma, or other malignancies. Another anti-inflammatory cytokine-therapy includes anti anti-IL 6R, anti-IL-12 or toxin-conjugated anti IL-7R, recombinant cytokines (IL-10 or IL 11). Given the diversity of proinflammatory products under its control, NF-kappaB may be viewed as a master switch in lymphocytes and macrophages, regulating inflammation and immunity. Although some of them still need more confirmatory studies, those immune therapies will provide new insights into cell-based and gene-based treatment against IBD in near future. PMID- 12769751 TI - Granulocyte colony-stimulating factor as the expecting sword for the treatment of severe sepsis. AB - Granulocyte colony-stimulating factor is a potent stimulator for neutrophils in the circulation as well as those progenitors to be increased and be grown in the bone marrow. Recombinant forms of this colony-stimulating factor, therefore, are widely using in the various kinds of clinical fields such as for the treatment of bone marrow suppression after cancer chemotherapy and for the treatment of myelodysplastic disorders. One of the physiological function of granulocyte colony-stimulating factor is to activate neutrophils enough to fight against invaded microbes and others, thus it is hoped to be used for the treatment of various kinds of infectious diseases even while the host has a normal number of circulating neutrophils and normal bone marrow. The future possible clinical usage of granulocyte colony-stimulating factor as for the immunomodulation against various kinds of insults to the human is reviewed. PMID- 12769752 TI - Pleiotropic effects of cytokines on acute myocardial infarction: G-CSF as a novel therapy for acute myocardial infarction. AB - Many cytokines have been reported to be increased in human and animal models with cardiovascular diseases. Myocardial infarction (MI) is accompanied with an inflammatory reaction which induces cardiac dysfunction and remodeling. The inflammatory reaction has been investigated in animal models of MI or myocardial ischemia-reperfusion injury. The mechanisms by which cytokine cascade is activated in the infarcted myocardium have been recently elucidated. Several hematopoietic growth factors including interleukin-3 (IL-3), IL-6, granulocyte macrophage colony-stimulating factors (GM-CSF), granulocyte colony-stimulating factor (G-CSF), and stem cell factor (SCF) have been reported to be positive regulators of granulopoiesis and act at different stages of myeloid cell development. G-CSF plays a critical role in regulation of proliferation, differentiation, and survival of myeloid progenitor cells. G-CSF also causes a marked increase in the release of hematopoietic stem cells (HSCs) into the peripheral blood circulation, a process termed mobilization. Although cardiac myocytes have been considered as terminally differentiated cells, it has been recently reported that there are many proliferating cardiac myocytes after MI in human heart. After it was demonstrated that bone marrow stem cells (BMSCs) can differentiate into cardiac myocytes, myocardial regeneration has been widely investigated. Recently, G-CSF has been reported to improve cardiac function and reduces mortality after acute MI. Although the mechanism by which G-CSF ameliorates cardiac dysfunction is not fully understood, there is the possibility that G-CSF may regenerate cardiac myocytes and blood vessels through mobilization of BMSCs. In the future, cytokine-mediated regeneration therapy may become to be a novel therapeutic strategy for MI. PMID- 12769753 TI - Therapeutically-induced autoantibodies in patients treated with recombinant hematopoietic growth factors: a brief summary. AB - Since recombinant gene technology was established to provide rare important regulatory proteins as recombinant molecules, cytokine therapies have widely developed and enormously contributed to the treatment of various diseases; nevertheless, it has been revealed that recombinant therapeutic molecules are not always perfect because of side-effects related to pharmacological functions of cytokines and/or potential antigenicity observed in some clinical cases. Although studies on the antigenicity of recombinant proteins have initiated, and observations in clinical studies have been accumulated over this decade, mechanisms of the autoantibody production are not clarified yet. Among various hematopoietic growth factors introduced into clinical trials, this report summarizes current issues of autoantibodies to primary regulators for terminal hematopoiesis. PMID- 12769754 TI - Clinical trials targeting advanced cancers by active immunization of T-cell defined tumor antigens. AB - Identification of CD8(+) and CD4(+) T-cell defined tumor antigens has opened the way for possible cancer immunotherapy. Peptide-based active immunization has begun mainly targeting melanoma. Surprisingly, some objective responses were obtained in the initial trials, leading to the accumulation of an explosive number of such antigens. Serological screening of cDNA expression libraries accelerated the identification of immunodominant antigens, resulting in the emergence of the concept of a "cancer-testis antigen." On the other hand, an advance in basic immunology has been promptly adopted: dendritic cells, as potent professional antigen presenting cells have been preferentially used in combination with the antigen in clinical trials both in vivo and in vitro. Tetramer analysis provided a powerful tool to directly monitor antigen-specific T cells in vivo in these trials. Moreover, adoptive transfer of antigen-specific T cells has added a new perspective to recent cancer immunotherapy. PMID- 12769755 TI - Clinical aspects of Clara cell 10-kDa protein/ uteroglobin (secretoglobin 1A1). AB - Clara cell 10-kDa protein (CC10)/ uteroglobin (UG) is a nonglycoprotein with a molecular mass of 16 kilodaltons, which is produced by mucosal epithelial cells in the lung (Clara cells), uterus and prostate. Like other low molecular weigh proteins it is catabolized in renal proximal tubules. Structurally it is a homodimer of subunits of 70 amino acids covalently bound in an antiparallel manner. It belongs to secretogobin (SCGB) family and is assigned as subgroup 1A1. The function of the protein so far elucidated is immunoregulatory and anti inflammatory in innate immunity. The knockout mouse of UG gene resulted in aggravation of inflammation by allergic and hyperoxic stimuli. It also showed very similar pathological features with human IgA nephropathy. The value is changed in the lung fluid and serum of various inflammatory and allergic lung diseases. Several kinds of single nucleotide polymorphisms (SNPs) in human CC10/UG gene were recently discovered; Adenine allele accumulation in G38A SNP has possible association with asthma and IgA nephropathy, being paralleled with disease severity of IgA nephropathy. Its expression is enhanced by some transcriptional factors induced by cytokines such as interferon-gamma. For cancer cells, the protein functions as an antagonist of neoplastic phenotype. CC10/UG forms one of intra- and intercellular regulators involved in inflammation and malignant transformation in the respiratory and urogenital fields. PMID- 12769760 TI - Application of microbial biotransformation for the new drug discovery using natural drugs as substrates. AB - This review describes microbial transformation studies of natural drugs, its importance in correlating them with the corresponding metabolism (biotransformation) in animal system and in the structural modification of complex drug molecules, which are difficult to obtain synthetically. Emphasis is laid on the potential microorganisms, which mimic the pathways of mammalian metabolism and which involve in stereoselective, regiospecific and selective conversions of natural drugs to their derivatives by microorganisms, which are more useful and difficult to synthesize. This paper describes the present state of-art, will elucidate the additional possibilities with microbial biotransformation of natural drugs, i.e., comparison of microbial transformation with mammalian metabolism and possibility of producing novel molecules, which are difficult to produce by other means. PMID- 12769761 TI - Hepatitis C virus clearance: the enigma of failure despite an impeccable survival strategy. AB - Infection with human hepatitis C virus (HCV) as a result of a bilateral process of host-virus interactions. There are factors on both sides that contribute to clearance and to chronicity. Virus strategy to survive is built on several basic features. The first, recently recognized, is a wide cell tropism. HCV can infect not only hepatocytes, but also cells of immune system (B-cells, monocytes, macrophages, dendritic cells), epithelium, and immunologically privileged sites such as the central nervous system. Dendritic cells and platelets can also be passive virus carriers. Possibilities of virus clearance or abortive inapparent HCV infection at the stage of adsorption are considered. The second feature is rapid error-prone replication that leads to accumulation within one host of multiple virus variants (quasispecies). Viral heterogeneity could be multiplied by recombination of HCV genomic/subgenomic RNA molecules. Quasispecies nature gives virus an advantage in adaptation to varying host environment including availability of permissive cells, the presence of innate and adaptive immune response, and antiviral treatment. Analysis of HCV polymorphisms and their evolution rates may pinpoint the molecular (sequence) correlates of HCV clearance. The third feature is the capacity to modify or adapt host milieu. HCV core, envelope E2 and nonstructural NS2, NS3, NS5A proteins seem to hold a grip over the host cellular functions by down-regulating processes unfavorable and up regulating processes favorable for virus replication and persistence. The relevance of the latter interactions to HCV infection outcome remains to be demonstrated. This review discusses recent developments in this area of HCV research. PMID- 12769762 TI - Host background factors contributing to hepatitis C virus clearance. AB - This review is an attempt to characterize the host in the earliest events of hepatitis C virus (HCV) infection before the on-set of adaptive immune response. Host meets the replicating HCV with innate immune response in the form of proinflammatory cytokine production, activation of natural killer (NK), NKT and dendritic cells. The potency of innate response is shaped by a wide panel of genetically predetermined constants and acquired variables. Higher rates of HCV clearance associate with white ethnicity and certain HLA haplotypes. Lower clearance rates correlate with genetic immune deficiencies/disorders. Recent findings link infection outcome with variation in the genes for the low-density lipoprotein and complement type 1 receptors. Important though insufficiently characterized is the role of polymorphisms in the genes responsible for induction of antiviral immunity. The outcome of HCV entry and of subsequent acute infection (if that occurs) is pre-determined by the immune competence of the host at the moment of infection. Higher rate of HCV clearance is observed for pediatric patients and young adults. Bad prognostic markers would be post-transplantation immune suppression, transfusion-related immune modulation, alcohol-induced immune imbalance and intoxication. Among host variables is the immune modulation induced by parasitic and viral co-infections. Some of the variables are transient and hard to define in retrospective. These host characteristics set up the potency, kinetics, and profile (Th1/Th2) of subsequent adaptive immune response. Better understanding of the host correlates of viral clearance would be a step towards prophylaxis of infection and an efficient anti-HCV vaccine. PMID- 12769763 TI - Invasive and non-invasive monitoring of hepatitis C virus-induced liver fibrosis: alternatives or complements? AB - Chronic hepatitis C virus (HCV) infection results in the development of liver fibrosis and cirrhosis in 20 to 25% of patients. The main task of the physician when examining a patient with a verified HCV infection is to identify the activity of inflammatory and necrotic processes in the liver, as well as the stage of fibrosis, and the reversibility of detected changes. Along with other clinical and laboratory parameters, this plays a major role in forecasting the course of hepatitis, as well as determines the therapeutic approach in each specific case. Liver biopsy remains the best way to assess the severity of chronic hepatitis C. The risk of developing cirrhosis depends on the stage (degree of fibrosis) and the grade (degree of inflammation and necrosis) observed in the initial liver biopsy. Non-invasive diagnostic approaches attempt to evaluate the serum markers of fibrogenesis. Biochemical markers of fibrosis scoring include thrombocyte counts, the prothrombin time, ratio of alaninaminotransferase (ALT) and aspartataminotransferase (AST) levels, the level of g-glutamyl transferase and the quantity of blood serum albumin. Another set of markers is based on the detection of molecular junctions that activate fibrosis, or participate in the generation of the liver extracellular matrix. The most applicable include hyaluronic acid (HA), type IV collagen (IV-C), N-terminal propeptide of type III procollagen (PIIIP), metalloproteinases (MMP), inhibitors of metalloproteinases (TIMP), and growth-transforming factor betta (GTFbeta). The review discusses the clinical significance of each of the criteria and possibility of their combination in the non-invasive monitoring of liver fibrosis. PMID- 12769764 TI - Carnitine metabolism and deficit--when supplementation is necessary? AB - Carnitine is an ammo acid derivative found in high energy demanding tissues (skeletal muscles, myocardium, the liver and the suprarenal glands). It is essential for the intermediary metabolism of fatty acids. Carnitine is indispensable for beta-oxidation of long-chain fatty acids in the mitochondria but also regulates CoA concentration and removal of the produced acyl groups. AcylCoAs act as restraining factor for several enzymes participating in intermediary metabolism. Transformation of AcylCoA into acylcarnitine is an important system for removing the toxic acyl groups. Although primary deficiency is unusual, depletion due to secondary causes, such as a disease or a medication side effect, can occur. Primary carnitine deficiency is caused by a defect in plasma membrane carnitine transporter in muscle and kidneys. Secondary carnitine deficiency is associated with several inborn errors of metabolism and acquired medical or iatrogenic conditions, for example in patients under valproate and zidovuline treatment. In cirrhosis and chronic renal failure, carnitine biosynthesis is impaired or carnitine is lost during hemodialysis. Other chronic conditions like diabetes mellitus, heart failure, Alzheimer disease may cause carnitine deficiency also observed in conditions with increased catabolism as in critical illness. Preterm neonates develop carnitine deficiency due to impaired proximal renal tubule carnitine re-absorption and immature carnitine biosynthesis. Carnitine stabilizes the cellular membrane and raises red blood cell osmotic resistance but has no metabolic influence on lipids in dialysis patients. L-Carnitine has been administered in senile dementia, metabolic nerve diseases, in HIV infection, tuberculosis, myopathies, cardiomyopathies, renal failure anemia and included in baby foods and milk. PMID- 12769771 TI - Death receptor signaling in cancer therapy. AB - Apoptosis, the cell's intrinsic death program, is a key regulator of tissue homeostasis. An imbalance between cell death and proliferation may result in tumor formation. Also, killing of cancer cells by cytotoxic therapies, such as chemotherapy, gamma-irradiation or ligation of death receptors is predominantly mediated by triggering apoptosis in target cells. Death receptor signaling pathways have been implied to contribute to the efficacy of cancer therapy. Failure to undergo apoptosis in response to anticancer therapy may lead to resistance. Understanding the molecular events that regulate apoptosis induced by anticancer therapy and how cancer cells evade apoptosis may provide new opportunities for drug development. Thus, novel strategies targeting tumor cell resistance will be based on insights into the molecular mechanisms of cell death. PMID- 12769772 TI - Implication of raft microdomains in drug induced apoptosis. AB - DNA damaging agents such as 1-beta-D-arabinofuranosylcytosine (Ara-C) and daunorubicin (DNR) are widely used in the treatment of acute nonlymphocytic leukemia. These drugs have, of course, been the objects of intense basic research, as well as preclinical and clinical study. Although specific biochemical lesions (DNA damage) have been associated with Ara-C- and DNR mediated cytotoxicity, the pathways leading to the induction of apoptosis remain ill defined. This standpoint has forced investigators to explore a new concept in cell response to cytotoxic stress: apoptosis signaling. The recent identification of a ceramide (CER) mediated apoptotic signaling pathway triggered by antitumor agents offers a new perspective for the treatment of neoplastic cells. Indeed, these agents have been shown to induce apoptosis through the activation of a sphingomyelinase (SMase) responsible for the hydrolysis of sphingomyelin (SM) and the generation of CER. The latter acts as a potent apoptosis mediator, triggering several downstream signaling pathways among which the stress-activated protein kinase cascade (MEKK1-SEK1-SAP/JNK) plays a critical role in apoptosis induction. However, the spacio-temporal organization of the key early signaling events is unclear. The present review delineates what appears to be a critical factor in apoptosis signaling: sphingomyelin enriched plasma membrane rafts. The apparent topological partitioning between DNA damage and apoptosis signaling (integrated into specialized plasma membrane domains) is discussed. PMID- 12769773 TI - Apoptosis induced by topoisomerase inhibitors. AB - Topoisomerase inhibitors are among the most efficient inducers of apoptosis. The main pathways leading from topoisomerase-mediated DNA damage to cell death involve activation of caspases in the cytoplasm by proapoptotic molecules released from mitochondria. In some cells, apoptotic response also involves the death receptor Fas (APO-1/CD95). The engagement of these apoptotic effector pathways is tightly controlled by upstream regulatory pathways that respond to DNA lesions-induced by topoisomerase inhibitors in cells undergoing apoptosis. These include the proapoptotic Chk2, c-Abl and SAPK/JNK pathways, the survival PI(3)kinase-Akt-dependent pathway and the transcription factors p53 and NF kappaB. Initiation of cellular responses to DNA lesions-induced by topoisomerase inhibitors is ensured by the protein kinases DNA-PK, ATM and ATR, which bind to DNA breaks. These kinases commonly called "DNA sensors" mediate their effects (DNA repair, cell cycle arrest and/or apoptosis) by phosphorylating a large number of substrates, including several downstream kinases such as c-Abl and the checkpoint protein Chk2. c-Abl induces apoptosis by activating cell death pathways (e.g., SAPK, p53 and p73) and inhibiting cell survival pathways [e.g., PI(3)kinase]. The DNA-damage regulating kinase Chk2, in addition to its role in cell cycle arrest and/or DNA repair, can induce apoptosis by phosphorylation/activation of the promyelocytic leukemia (PML) protein and p53. Finally, we will review the recent observations that support a role for topoisomerases in chromatin fragmentation during the execution phase of apoptosis. PMID- 12769774 TI - Signal transduction pathways of taxanes-induced apoptosis. AB - Docetaxel (Taxotere) is a member of the taxane class of anticancer agents to reach clinical use. This semisynthetic analog of paclitaxel (Taxol) is one of the newer potent anti-neoplastic agents now undergoing extensive laboratory and clinical investigations. Several studies indicate that antimicrotubule agents are potent promoters of apoptosis in cancer cells. Cytotoxic mechanisms of antimitotic taxoids are not yet fully understood, but it has been demonstrated that docetaxel increases tubulin polymerisation, promotes microtubule assembly and also inhibits tubulin depolymerisation. Disruption of microtubules results also in the induction of tumor suppressor gene p53 and inhibitor of cyclin dependent kinases and activation/inactivation of several protein kinases. As a consequence cells are arrested in the G2-M phase of the cell cycle, after which they may either undergo cell death by apoptosis or necrosis or overcome the G2-M stop and continue in the division cycle (often toward a post-mitotic cell death) depending on the tumor cell type. Nevertheless, how docetaxel induces apoptotic cell death or caspases activation is not yet defined. One may assume that taxanes are able to induce the phosphorylation of Bcl-X(L)/Bcl-2 members and thus inactivate their anti-apoptotic capacities. The down-regulation of Bcl-2 and/or the upregulation of p53 and p21/WAF-1 are certainly one of the important modes of apoptosis induction by taxanes. The aim of this framework is to summarize the effects of microtubuline targeting agents on apoptotic signal transduction and new molecular pathways. Finally, we will also discuss the potential therapeutic interest in the association of docetaxel and ionizing radiation. PMID- 12769775 TI - Bcl-2 proteins: targets and tools for chemosensitisation of tumor cells. AB - Proteins of the Bcl-2 family share one or several Bcl-2 homology (BH) regions and behave as pro- or anti-apoptotic proteins. Prosurvival members such as Bcl-2 and Bcl-X(L) are supposed to preserve mitochondrial outer membrane integrity, thus preventing the release of soluble apoptogenic molecules. Pro-apoptotic members include BH3-only proteins that act as sensors of cellular damage and initiate the death process and Bax-like proteins that act downstream of BH3-only proteins to permeabilise the mitochondrial outer membrane. Whether BH3-only proteins directly activate Bax-like proteins or prevent prosurvival members of the family from inhibiting Bax-like proteins or both remains a matter of controversy. Expression of these proteins is altered in various human tumours and this abnormal expression may contribute to oncogenesis and tumour cell resistance to anticancer drug-induced cell death. Based on these observations, prosurvival proteins are attractive intracellular targets for inducing tumour cell death or sensitising tumour cells to death induced by chemotherapeutic drugs. The use of 18-mer antisense oligonucleotides (G3139 or Genasense) targeting the first six codons of bcl-2 mRNA is currently developed in clinics with phase I studies demonstrating that thrombocytopenia may be the main dose-limiting side effect. This strategy, that efficiently decreases Bcl-2 protein expression in some tumour cells, is currently tested in phase II and phase III trials. Alternative approaches to achieve the functional knock-out of Bcl-2 include the use of either peptides mimicking the BH3 domain of Bcl-2-related proteins or more stable, non peptidic BH3 mimetics and the pharmacological modulation of the post-translational modifications of the protein. PMID- 12769776 TI - Drugs acting on the beta isoform of human topoisomerase II (p180). AB - Topoisomerase II is the target of several anticancer agents. The discovery of a second enzyme, called topoisomerase II beta, genetically distinct from alpha, prompted the investigation on the different functional roles of the two isoforms. Whereas the first recognized isozyme is essential for life due to its role in chromosome condensation and segregation, beta functions remained elusive, although its importance in neural development is appearing clearer. Topoisomerase II beta is regulated differently than alpha, and its level of expression does not change significantly during cell cycle. The presence of this isoform in non proliferating cells suggests that drug preferentially aimed at beta may be active in slow growing tumors. Topoisomerase II poisons were hence evaluated in light of their selectivity toward one or the other isozyme, indicating how the beta isoform may represent an important target for selected classes of drugs. Newer compounds were also synthesized and tested for their potential antitumor activity and their topoisomerase II beta poisoning. The literature dealing with "old" and "new" drugs targeted at topoisomerase II is reviewed trying to link, whenever possible, selective poisoning and cytotoxic effects to chemical structures, in the hope to indicate new lead compounds that will contribute to unveil molecular determinants of selectivity. PMID- 12769777 TI - The interaction of histone deacetylase inhibitors and DNA methyltransferase inhibitors in the treatment of human cancer cells. AB - The potential anticancer activities of histone deacetylase (HDAC) inhibitors and DNA methyltransferase (DNMT) inhibitors have been extensively studied in recent years. HDAC inhibitors suppress the activities of multiple HDACs, leading to an increase in histone acetylation. This histone acetylation induces an enhancement of the expression of specific genes that elicit extensive cellular morphologic and metabolic changes, such as growth arrest, differentiation and apoptosis. DNMT inhibitors, such as 5-aza-cytidine (5-aza-CR) and 5-aza-2'-deoxycytidine (5-aza CdR) are also widely studied because DNA hypomethylation induces the re activation of tumor suppressor genes that are silenced by methylation-mediated mechanisms. Recently, the combination of HDAC inhibitors or demethylating agents with other chemo-therapeutics has gained increasing interest as a possible molecularly targeted therapeutic strategy. In particular, the combination of HDAC inhibitors with demethylating agents has become attractive since histones are connected to DNA by both physical and functional interactions. To date, the accumulating evidence has confirmed the hypothesis that the combination of HDAC and DNMT inhibition is very effective (and synergistic) in inducing apoptosis, differentiation and/or cell growth arrest in human lung, breast, thoracic, leukemia and colon cancer cell lines. This review will discuss the in vitro effects of HDAC inhibitors, such as trichostatin A (TSA), sodium butyrate, depsipeptide (FR901228, FK228), valproic acid (VPA) and suberoylanilide hydroxamic acid (SAHA), and the demethylating agent, 5-aza-CdR used alone and in combination treatment of human cancer cells and the possible mechanisms involved. PMID- 12769778 TI - Signal transduction inhibitors in the treatment of breast cancer. AB - Signal transduction pathways are frequently altered in human breast cancer and are the targets of several novel therapies currently in clinical trials. Therapeutic strategies include extracellular blockade of tyrosine kinase receptors with the monoclonal antibodies C225 and trastuzumab. Competitive inhibitors of adenosine triphosphate binding sites on tyrosine and serine/threonine kinases are also being evaluated in phase I/II trials; these include ZD1839, OSI-774 and CI-1033. Flavopiridol and UCN-01 are nonspecific cell cycle kinase antagonists with preliminary evidence of breast cancer cell growth inhibition. Several inhibitors of mitogen-activated protein kinase and phosphatidylinositol 3-kinase signaling are also in various stages of preclinical or clinical development. Additionally, inhibitors of farnesyl transferase have demonstrated activity in breast cancer cells irrespective of ras status. Current evidence suggests that targeting of signaling molecules is a promising new approach to treatment of breast cancer. PMID- 12769779 TI - The BCL2-family of protein ligands as cancer drugs: the next generation of therapeutics. AB - Selective aberrant cell suicide (ie., apoptosis or programmed cell death) is a hallmark of "nonneoplastic" tissue. In cells that have clonally evolved or in common parlance "cancer cells", apoptosis is either itself aberrant or completely inhibited. Strategies to enhance apoptosis under conditions of cancer cellular stress is an evolving and actively investigated area of experimental therapeutics. Bcl2 proteins are key mediators of the process of apoptosis and ligands to these family of proteins have been described using modern combinatorial, computational and evolutionary small molecule screening approaches. Crystallization of several of the Bcl2 family members has provided clarification of the role of these ligands and provided a clearer mechanism of action for the consequences of ligand binding. In several cases, these ligands (e.g., HA14-1, 2-methoxy antimycin A) induce apoptosis even under conditions of Bcl2 overexpression and if developed preclinically will be promising anticancer agents. This rationale becomes even more striking when one observes overexpression of Bcl2 in 70% of breast cancer, 30-60% of prostate cancer, 80% of B-cell lymphomas, 90% of colorectal adenocarcinomas, and many other forms of cancer. PMID- 12769780 TI - Pharmacogenetics of irinotecan. AB - Pharmacogenetics focuses on intersubjects variation in therapeutic drug effects and toxicity depending on genetic polymorphisms. This is particularly interesting in oncology since anticancer drugs usually have a narrow margin of safety. Irinotecan [7-ethyl-10-[4-(1-piperidino)-1-piperidino]carbonyloxycamptothecin] is used in cancer chemotherapy as a topoisomerase I inhibitor and it is characterised by a sometimes unpredictable severe toxicity. It is mostly intestinal with nausea, vomit and diarrhoea or haematologic with leuko thrombocytopenia. Its complex metabolism involves many proteins. Human carboxylesterase isoforms 1 and 2 (hCE1, hCE2) activate irinotecan to its metabolite SN-38 (7-ethyl-10-hydroxycamptothecin); cytochrome P450 isoforms 3A4 and 3A5 (CYP3A4, CYP3A5) mediate the oxidation of the parental compound to irinotecan; uridino-glucuronosil transferase isoform 1A1 (UGT1A1) catalyses glucuronidation of SN-38; the multi-resistance protein isoform 2 (MRP2) allows the cellular excretion of the SN-38 glucuronide (SN-38G) and the multi-drug resistance gene (MDR1), encoding for P-glycoprotein, is responsible for the excretion of irinotecan from the cell. Polymorphic structures in the genes encoding for all these proteins have been described. In particular, the UGT1A1*28 allele has been associated with an increased toxicity after irinotecan chemotherapy. Classical parameters used in the clinic, such as body-surface area, have no longer a meaningful correlation with clinical outcome. Hence it emerges the importance of studying the individual genotype to predict the toxicity and efficacy of irinotecan and to individualise therapy. In this review, we summarise the new developments on the study of the pharmacogenetics of irinotecan, stressing its importance in drug cytotoxic effect. PMID- 12769781 TI - Peroxisome proliferator activated receptor-gamma ligands as potent antineoplastic agents. AB - The Peroxisome Proliferator Activated Receptors (PPARs) are initially described as molecular targets for compounds inducing peroxisome proliferation. Among the three PPAR subtypes (alpha, beta, gamma), PPAR-gamma acting as a ligand-activated transcription factor, proved to be an important regulator of adipogenic differentiation and glucose homeostasis. Recent data support evidence for participation of PPAR-gamma, upon ligands activation, in the biological mechanisms underlying the carcinogenic evolution. Specific PPAR-gamma ligands affect cancer cells proliferation and differentiation acting as cell cycle modulators, suggesting their use as an important tool for future therapeutic approach in cancer. In this review, the latest knowledge on PPAR-gamma activation and molecular mechanisms of PPAR-gamma ligands mediated anti-tumoral activity are presented. In vitro and in vivo studies concerning the use of PPAR-gamma ligands in different cancer types are also included. PMID- 12769782 TI - Biosynthetic pathways of plastid-derived organelles as potential drug targets against parasitic apicomplexa. AB - Apicomplexan parasites are a large phylum of unicellular and obligate intracellular organisms of great medical importance. They include the human pathogens Plasmodium spp., the causative agent of malaria, and Toxoplasma gondii, an opportunistic parasite of immunosuppressed individuals and a common cause of congenital disease, together affecting several hundred million people worldwide. The search for new and effective drugs against these pathogens has been boosted during the last years by an unexpected finding. Through molecular and cell biological analysis it was realized that probably most members of this phylum harbor a plastid-like organelle, called the apicoplast, which probably is derived from the engulfment of a red alga in ancient times. Although the apicoplast itself contains a small circular genome, most of the proteome of this organelle is encoded in the nuclear genome, and the proteins are subsequently transported to the apicoplast. It is assumed to contain a number of unique metabolic pathways not found in the vertebrate host, making it an ideal "playground" for those interested in drug targets. Recent reports have shown that the rationale of this approach is valid and that new drugs which are urgently needed especially for plasmodial infections, might be developed in the near future based on these targets. Amongst them are three enzymes of the plant-like fatty acid synthesis machinery and enzymes of the non-mevalonat isoprenoid biosynthesis pathway. From their presence in the apicoplast it can be concluded that fatty acid and lipid biosynthesis seems to be a major function of the apicoplast. Another recently described apicoplast enzyme, ferredoxin-NADP(+)-reductase and its redox partner, ferredoxin, points to another interesting organelle-specific biosynthetic pathway, namely [Fe-S] cluster biosynthesis. In the present review, the fundamental aspects of the apicoplast as drug target will be described, together with the specific pathways and their currently known inhibitors. Furthermore, based on the recent findings potentially new targets will be discussed. A short overview of the presently available high-throughput methods for Apicomplexa to evaluate the potency of new inhibitory substances will also be given. PMID- 12769783 TI - AIDS-related insulin resistance and lipodystrophy syndrome. AB - The recent development of highly active antiretroviral therapy (HAART) has drastically improved the life expectancy of AIDS patients, by reducing infection related mortality. However, the prolongation of the lives of HIV-1-infected patients and/or the long-term use of novel, potent antiviral agents have generated a score of new problems and complications. Among them is the AIDS related insulin resistance and lipodystrophy syndrome, which is observed in 30 80% of AIDS patients who are well controlled by HAART. This syndrome is associated with severe metabolic disturbances, such as carbohydrate intolerance/diabetes mellitus and dyslipidemia, which cause atherosclerotic cardiovascular disease. The etiology of this syndrome appears to be multi factorial; other than the anti-viral drugs, hypercytokinemia and the HIV-1 infection itself, including the virally encoded molecules Vpr and Tat, could contribute to the development of these pathologic changes or increase the vulnerability of patients to the adverse effect of the therapeutic compounds. In this article, we review our current understanding of the pathogenesis and therapeutic approach of this newly emerging AIDS-associated metabolic syndrome. PMID- 12769784 TI - Surface-linked liposomal antigen induces IgE selective unresponsiveness in a T cell independent fashion. AB - We previously reported that surface-linked liposomal antigen induced IgE selective unresponsiveness. The results were consistent even when different coupling procedures for antigen with liposomes, or for liposomes with different lipid components, were employed. During the course of an investigation intended to clarify the mechanism of IgE-selective unresponsiveness induced by surface coupled liposomal antigens, we discovered an alternative approach to regulate the production of IgE, one that is independent of the activity of T-cells. Immunization of mice with OVA-liposome conjugates induced IgE- selective unresponsiveness without apparent Th1 polarization. Neither interleukin-12 (IL 12), IL-10, nor CD8(+) T-cells participated in the regulation. Further, CD4(+) T cells of mice immunized with OVA-liposome were capable of inducing antigen specific IgE synthesis in athymic nude mice immunized with alum-adsorbed OVA. On the other hand, immunization of the recipient mice with OVA-liposome did not induce anti-OVA IgE production, even when CD4(+) T-cells of mice immunized with alum-adsorbed OVA were transferred. In the secondary immune response, OVA liposome enhanced anti-OVA IgG antibody production but not the ongoing IgE production, suggesting that the IgE-selective unresponsiveness induced by the liposomal antigen involved direct effects on IgE but not IgG switching in vivo. These results suggest the role of an alternative mechanism, one not involving T cells, in the regulation of IgE synthesis, and raise the possibility that surface linked liposomal antigen is potentially applicable for the development of a novel vaccine that induces the least IgE synthesis. Moreover, given the relatively low allergic response to and increased antigenicity of the allergen, this form of antigen preparation would be applicable to allergen immunotherapy. PMID- 12769785 TI - The evolving therapeutic approaches for Epstein-Barr virus infection in immunocompetent and immunocompromised individuals. AB - Epstein-Barr virus (EBV) is one of eight known human herpesviruses (HHVs). A primary EBV infection is generally subclinical in immunocompetent individuals, but often causes infectious mononucleosis (IM) in adolescents and adults, which is generally a benign and self-limiting disease. Therefore, in immunocompetent individuals only symptomatic treatment is recommended, although fatal or malignant diseases such as fatal IM, Burkitt's lymphoma (BL) and nasopharyngeal carcinoma (NPC) may develop without obvious preceding immunodeficiency. However, in certain circumstances such as in patients with hereditary immunodeficiencies, in recipients receiving a potent immunosuppressant or in patients with acquired immunodeficiency syndrome (AIDS), this virus strongly links to the development of lethal lymphoproliferative diseases (LPD). These LPD range from IM-like illness associated with polyclonal proliferation to malignant lymphoma in monoclonal fashion. To date, no specific therapy has been available for latent EBV infection itself, but understanding the underlying pathogenetic mechanisms in each condition provides the possible treatment including anti-viral agents, immune modulators and chemotherapeutic drugs. Furthermore, severe combined immunodeficiency (SCID) mouse engrafted with human peripheral blood mononuclear cells is a suitable model for EBV-associated LPD which occur in human beings. Using this, several therapeutic trials have been investigated, and some are possibly beneficial. This concise review focuses on recent understanding of the pathogenetic mechanisms in various EBV-associated diseases in immunocompetent and immunocompromised individuals, and discusses potent therapeutic approaches in each condition. PMID- 12769786 TI - Drug-induced aseptic meningitis. AB - Aseptic meningitis is a rare but well-recognized complication of drug therapy. The clinical presentation of drug-induced aseptic meningitis (DIAM) is distinct. Symptoms typically include fever, neck stiffness, headache, confusion, nausea and vomiting. The major categories of causative agents are non-steroidal anti inflammatory drugs, antimicrobials and also intravenous immunoglobulins, monoclonal antibodies and vaccines. These drugs most commonly implicated as causes of aseptic meningitis act more likely through an immunological mechanisms. However, the exact pathogenetic mechanism of DIAM is still unknown. The diagnosis of drug-induced aseptic meningitis is difficult and infectious etiologies must be excluded. In some cases the diagnosis has been confirmed by rechallenging the patient with the suspected agent. In this case, informed written consent is necessary and rechallenge must be medically supervised both to document the response and to offer medical care and advice, if required. The outcome of DIAM is generally good, usually without long term sequelae. PMID- 12769787 TI - Metformin and its liver targets in the treatment of type 2 diabetes. AB - Although a number of assessments disagree, the preponderance of the evidence indicates that the major therapeutic action of metformin in type 2 diabetes (DM2) is on the liver, and glucose production (EGP) in particular. At the level of this organ, the actions of metformin can be characterized as pleiotropic. The major questions addressed here are therefore: (i) the methodological aspects of the determination of glucose fluxes: when glucose production is not found to be elevated in type 2 diabetes, it is not surprising that little action of metformin on this flux is found. The issues of populations examined, experimental protocols, and quantitative methods of flux determination are important in answering this question. Early morning EGP is increased and constitutes a valid target for metformin. (ii) the multiple targets of metformin: metformin acts at a number of sites and interacts with metabolites and hormones. Some of these actions may be expressed at different doses. Although their net effect is therapeutic, not all are oriented towards lowering hyperglycemia, perhaps explaining the more modest effect of this drug than could be anticipated from individual actions. Sites of metformin action can therefore be considered as a compilation of valid therapeutic targets in DM2. Gluconeogenesis, glycogenolysis and glycogen synthesis can be altered by metformin, although in vivo, this also depends on the methodology. Component processes from substrate supply and liver uptake, through a number of glucogenic enzymes, as well as glycogen synthase and phosphorylase have all been shown to be affected. (iii) unifying concepts: reported actions of metformin on the mitochondrial respiratory chain, free fatty acid metabolism, AMP-activated protein kinase, and on membrane proteins directly may all explain subsets of actions that are seen, providing more integrated targets for consideration in the therapy of DM2. PMID- 12769788 TI - CD8+ T-cell-mediated non-cytolytic suppression of human immuno-deficiency viruses. AB - Major histocompatibility (MHC)-restricted cytotoxic T-lymphocytes (CTL) kill human immunodeficiency virus (HIV)-infected cells. In addition, activated CD8(+) T-lymphocytes from HIV-infected individuals suppress virus replication in vitro by producing antiviral factor (CAF). The effector mechanism(s) of CAF involves modulation of HIV gene transcription, is non-cytolytic and mediated in part by soluble antiviral factors. Initially, CAF activity was shown to be more vigorous in activated CD8(+) cells and cell free supernatants (SNs) from asymptomatic individuals compared to those with AIDS, suggesting a protective role in vivo. CAF-mediated suppression is also evident in animal models of immunodeficiency virus infection. Several soluble molecules that contribute to non-cytolytic virus suppression have been characterised, including alpha- and beta-chemokines and interleukin-16 (IL-16), but these are distinct from CAF. Two agents possessing certain CAF-like characteristics, modified antithrombin 111 (AT111) and the human alpha-defensins, have been described but their antiviral mechanisms are not fully understood. CAF-secretion may not be virus-specific as similar activity is found in activated CD8(+) cells/SNs from humans and chimpanzees seronegative for HIV-1. Recent data indicates that the secretion of CAF is MHC-restricted and both cytolytic and non-cytolytic mechanisms are mediated by CTL. If the latter is correct, a single appropriate stimulus could be used to enhance both effector mechanisms in vivo. This paper reviews research aimed at characterising HIV suppressive factors and raises other questions that must be considered for the development of prophylactic and therapeutic strategies leading to the safe and effective control of HIV. PMID- 12769789 TI - Targeted inhibition of angiogenic factors in AIDS-related disorders. AB - Immunosuppression associated with human immunodeficiency virus (HIV) infection and acquired immunodeficiency syndrome (AIDS) markedly increases the risk for development of several cancers. Despite its dramatic decrease in frequency after the introduction of highly active antiretroviral therapy (HAART), Kaposi's sarcoma (KS) remains the most common neoplastic manifestation of AIDS. KS is a multicentric angioproliferative tumor, characterized microscopically by spindle cells. KS cells produce and respond to angiogenic factors such as basic fibroblast growth factor (bFGF) and vascular endothelial growth factor-A (VEGF A). In addition to cellular growth factors, the trans-activator HIV protein Tat plays a major role in the pathogenesis of AIDS-related KS by augmenting the angiogenic activities of bFGF and VEGF-A, and activating the VEGF receptor-2. Viral products from the recently described Kaposi's sarcoma-associated herpesvirus (KSHV) also exhibit potent angiogenic activities. KSHV is consistently associated with KS and two lymphoproliferative disorders, primary effusion lymphoma (PEL) and the plasma cell variant of multicentric Castleman's disease (MCD). Several viral genes may contribute to the phenotype of PEL and MCD: among them, a viral homologue of interleukin-6 (vIL-6) has attracted much attention due to its potential to stimulate B cell growth and accelerate angiogenesis via VEGF-A induction. In this review, we summarize current knowledge and hypothesis regarding the cellular and viral angiogenic factors involved in the pathogeneses of AIDS-related malignancies, and discuss novel therapeutic strategies based on targeting pro-angiogenic factors. PMID- 12769790 TI - Viral latent proteins as targets for Kaposi's sarcoma and Kaposi's sarcoma associated herpesvirus (KSHV/HHV-8) induced lymphoma. AB - Kaposi's sarcoma-associated herpesvirus (KSHV/HHV-8) is present in all Kaposi's sarcoma tumor cells as well as in several lymphomas that are linked to this agent. Every tumor cell expresses the viral latent protein LANA, which is required for KSHV latent replication and proper segregation of the viral episome. In certain tumors, other latent KSHV proteins (LANA-2/vIRF3, v-cyclin, v-IL6) are expressed as well. Since all herpesviruses persist for life in infected individuals, only eradication of latent virus can cure infection. The KSHV latent genes serve as bona fide tumor markers, but do they also provide targets for anti tumor and/or anti-viral drugs? To decide this question we review the known biochemical interactions between KSHV latent proteins and their viral and cellular partners. Recent epidemiological studies show that KSHV lytic replication precedes KSHV associated cancers. Gancilovir has been linked to KS tumor regression, which implicates the KSHV-encoded polymerase as a potential intervention point. Yet, KSHV specific transactivators might represent more specific targets, as they have no cellular homologs. In particular Rta/orf50 is necessary and sufficient for lytic replication and deserves serious consideration as a target for KSHV-specific antivirals. PMID- 12769791 TI - Current perspectives on cytokines for anti-retroviral therapy in AIDS related B cell lymphomas. AB - The development of high-grade B-cell lymphoma in Acquired Immunodeficiency Syndrome (AIDS) patients is a relatively late manifestation induced by Human Immunodeficiency Virus-1 (HIV) infection and is considered to be an AIDS-defining condition. Multiple, ongoing molecular and cytogenetic aberrations appear necessary for the development of AIDS-related lymphoma. Studying a panel of human B-cell lines derived from patients with Burkitt's lymphoma (BL) and AIDS associated Burkitt's lymphoma (AIDS-BL) we had described constitutive expression and secretion of large amounts of Interleukin-16 (IL-16), Macrophage Inflammatory Protein-1alpha (MIP-1alpha), Macrophage Inflammatory Protein-1beta (MIP-1beta), Interleukin-12 (IL-12), Interleukin-10 (IL-10), and Interleukin-7 (IL-7). Some of these cytokines like IL-16, MIP-1beta, MIP-1alpha and Regulated upon activation normal T expressed and secreted (RANTES) are shown to have inhibitory effect on HIV replication. Interestingly, we identified a novel transcription factor family, Macrophage Inflammatory Protein-1alpha Nuclear Protein (MNP), which is suggested as a potential target for anti-retroviral therapy based on the implication of its role and involvement as a key regulator of MIP-1alpha. It is apparent, that HIV induces the production of a cascade of cytokines and cytokine receptors. Some of these molecules serve to increase the infection and replication of HIV per se, and some others serve to induce a state of B-cell growth, activation, and differentiation. This review attempts to delineate the complex mechanisms of viral, B-cell, oncogene, cytokine/cytokine receptor and transcription factor interactions that are involved in AIDS associated lymphomagenesis. Unfolding the relationship between cytokines and the underlying mechanisms of the disease will not only help in understanding the pathophysiology but also will facilitate focusing on the development of new diagnostic and therapeutic strategies. PMID- 12769792 TI - Particle-based vaccines for HIV-1 infection. AB - The use of live-attenuated viruses as vaccines has been successful for the control of viral infections. However, the development of an effective vaccine against the human immunodeficiency virus (HIV) has proven to be a challenge. HIV infects cells of the immune system and results in a severe immunodeficiency. In addition, the ability of the virus to adapt to immune pressure and the ability to reside in an integrated form in host cells present hurdles for vaccinologists to overcome. A particle-based vaccine strategy has promise for eliciting high titer, long-lived, immune responses to a diverse number of viral epitopes from different HIV antigens. Live-attenuated viruses are effective at generating both cellular and humoral immunity, however, a live-attenuated vaccine for HIV is problematic. The possibility of a live-attenuated vaccine to revert to a pathogenic form or recombine with a wild-type or defective virus in an infected individual is a drawback to this approach. Therefore, these vaccines are currently only being tested in non-human primate models. Live-attenuated vaccines are effective in stimulating immunity, however challenged animals rarely clear viral infection and the degree of attenuation directly correlates with the protection of animals from disease. Another particle-based vaccine approach for HIV involves the use of virus-like particles (VLPs). VLPs mimic the viral particle without causing an immunodeficiency disease. HIV-like particles (HIV-LP) are defined as self assembling, non-replicating, nonpathogenic, genomeless particles that are similar in size and conformation to intact virions. A variety of VLPs for both HIV and SIV are currently in pre-clinical and clinical trials. This review focuses on the current knowledge regarding the immunogenicity and safety of particle-based vaccine strategies for HIV-1. PMID- 12769793 TI - Immunological approaches for HIV therapy. AB - The induction of a Th-1 polarized immune response is believed to be advantageous when designing immunologic approaches for HIV therapy. DNA vaccines represent one of the best immunologic strategies capable of inducing such a response. From conception to clinical application it is now possible to rationally design DNA vaccines based on reliable experimental data, thus a systemic approach to the development of new and the enhancement of existing vaccine immunogens is now possible. The addition of adjuvants may also increase immunogenicity and depending on the choice of adjuvant, polarize the immune response. Other important factors in the formulation of a successful vaccine are the selection of administration route, heterologous or homologous prime/boost schedules, and the feasibility of the eventual clinical application. This review will summarize recently developed preventive and therapeutic vaccines, and carefully evaluate the advantages and potential risks for Human Immunodeficiency Virus (HIV) infected patients. Finally, the concept of "autovaccination" will be defined as it represents the basis for the development of our innovative therapeutic antigen presenting cell targeted HIV vaccine. DermaVir is the first topical vaccine, in combination with antiretroviral therapy, to demonstrate immunological and clinical benefits in a relevant animal model (chronically infected rhesus macaques). PMID- 12769794 TI - Viral reservoirs an impediment to HAART: new strategies to eliminate HIV-1. AB - The introduction of potent drug combinations comprising reverse transcriptase and protease inhibitors has dramatically altered the natural history of HIV disease, at least in the short term. Unfortunately, poor penetrability into different anatomic compartments, toxicity and drug resistance, are some of the problems related to their prolonged use. HIV's ability to mutate and become resistant along with the ongoing viral replication during HAART, which may lead to the emergence of independently evolving viral strains in different anatomic compartments and establishment of latent viral reservoirs also remain critical for the success and failure of antiretroviral therapy. Current drug therapies do not eliminate these viral reservoirs, nor do they discourage their formation. New strategies are needed for flushing hidden pockets of HIV in vivo. This review will focus mainly on novel strategies in the pipeline, along with the recent developments in the field. PMID- 12769797 TI - Role of acetylcholinesterase inhibitors in the metabolism of amyloid precursor protein. AB - Potentiation of central cholinergic activity has been proposed as a therapeutic approach for improving the cognitive function in patients with Alzheimer's disease (AD). Increasing the acetylcholine concentration in the brain by modulating acetylcholine-sterase (AChE) activity is among the most promising therapeutic strategies. Efforts to treat the underlying pathology based on the modulation of amyloid precursor protein (APP) processing in order to decrease the accumulation of beta-amyloid are also very important. Alterations in APP metabolism have recently been proposed to play a key role in the long-lasting effects of AChE inhibitors. This review surveys recent data from in vivo and in vitro studies that have contributed to our understanding of the role of AChE inhibitors in APP processing. The regulatory mechanisms relating to the muscarinic agonist effect, protein kinase C activation and mitogen-activated protein kinase phosphorylation, involving the alpha-secretase or the 5 -UTR region of the APP gene, are also discussed. Further work is warranted to elucidate the exact roles in APP metabolism of the AChE inhibitors used in AD therapy at present. PMID- 12769795 TI - Recent knowledge on molecular components of Lewy bodies discloses future therapeutic strategies in Parkinson's disease. AB - Lewy bodies (LB) were first described by Lewy in 1912 [1] as neuronal pale eosinophilic inclusions which became a pathological hallmark of Parkinson s disease (PD). In his original study, Lewy defined these inclusions as pale eosinophilic cytoplasmic structures, and studies since then have revealed LB to be ubiquitin-, alpha-synuclein-, and parkin-containing inclusions. This suggests that knowledge of the biochemical steps involved in the genesis of LB might disclose a final common pathway which might be responsible for different types of inherited and sporadic parkinsonism. This would lead to the identification of new therapeutic targets for interfering with disease progression. Although LB were originally described solely in PD, in the last decade these inclusions were described in a spectrum of degenerative disorders ranging from pure movement disorders to dementia. This suggests that common biochemical alterations leading to the formation of intracellular inclusions might underlie various pathological conditions. Consequently, the knowledge of the biochemical steps involved in the formation of neuronal inclusions could represent a key to develop new therapeutic strategies. In recent years it has been possible to develop both in vitro and in vivo neuronal inclusions resembling Lewy bodies. These experimental approaches have ranged from the use of alpha-synuclein transgenic mice to the continuous exposure to a mitochondrial complex I inhibitor. The aim of the present paper is to review briefly, recent advances on Lewy body research to achieve new insight into the etiology of PD and the molecular events leading to neurodegeneration. PMID- 12769798 TI - Calpain and its involvement in the pathophysiology of CNS injuries and diseases: therapeutic potential of calpain inhibitors for prevention of neurodegeneration. AB - Calpain is a Ca(2+)-activated proteolytic enzyme involved in neurodegeneration in a variety of injuries and diseases of the central nervous system (CNS). Many calpain homologs have been discovered. Depending on the tissue distribution, calpains are broadly classified as ubiquitous and tissue-specific. Ubiquitous calpain isoforms, -calpain and m-calpain, are abundantly expressed in the CNS. Calpastatin, an endogenous protein inhibitor, regulates the activity of ubiquitous calpain. Overactivation of calpain may degrade calpastatin, limiting its regulatory efficiency. Molecular structures of calpain and calpastatin have been deduced from cDNA cloning. The precise physiological function of calpain remains elusive. However, experimental evidence strongly suggests an important role for calpain in causing neurodegeneration in various injuries and diseases of the CNS. The increase in intracellular free Ca(2+) levels in the course of injuries and diseases in the CNS causes overactivation of calpain, promoting degradation of key cytoskeletal and membrane proteins. Cleavage of these key proteins by calpain is an irreversible process that perturbs the integrity and stability of CNS cells, leading to programmed cell death or apoptosis. Calpain in conjunction with caspases can cause apoptosis of the CNS cells. An aberrant Ca(2+) homeostasis inevitably activates calpain, which plays a crucial role in the pathophysiology of the CNS injuries and diseases. Therefore, calpain is a potential therapeutic target to prevent neurodegeneration. To this end, various cell-permeable calpain inhibitors have been synthesized for pharmacological inhibition of calpain activity. Some calpain inhibitors have shown significant neuroprotection in animal models of the CNS injuries and diseases, indicating their therapeutic potential. PMID- 12769796 TI - FK506 and its analogs - therapeutic potential for neurological disorders. AB - Immunophilin ligands such as FK506 and Cyclosporin A, used in immunosuppression, are well-characterized drugs. In the past, they had been the center of attention as a putative therapeutic strategy for neuroregeneration and neuroprotection. In contrast to Cyclosporin A, FK506 readily crosses the brain-blood-barrier and, thus together with its derivatives, may represent a novel approach to the treatment of neurological disorders. FK506 exerts profound neuroprotective and neuroregenerative effects in vivo and in vitro. The mechanism underlying neuroregeneration is fairly well understood. It is independent of the inhibition of calcineurin, which is responsible for the immunosuppression, but operates via the binding of FKBP52 and the heat shock protein (Hsp) 90. In contrast, the underlying pathways of neuroprotection are far less understood. Protection is apparently independent of calcineurin, as shown by non-calcineurin inhibiting derivatives, such as V-10,367 and GPI-1046, but the intracellular actions remain to be defined. FK506 has been shown to interfere with the apoptotic pathway of neuronal cells, including inhibiting JNK activity, cytochrome c release, caspase 3 activation, and CD95 ligand expression. These effects are in part mediated by the inhibition of calcineurin and may not contribute to protection. Our recent studies suggest that the protective properties of FK506 and its non-calcineurin inhibiting derivatives are realized by a fast induction of heat shock proteins. The induction of the heat shock response by immunophilin ligands might prove to be an interesting target for neuroregeneration and neuroprotection. PMID- 12769799 TI - The vasopressin V1b receptor as a therapeutic target in stress-related disorders. AB - The complexity of the stress response would appear to provide multiple opportunities for intervention, but treatment strategies are often centered on the improvement of symptoms rather than attempting to "treat" the stress response. However, recent efforts have begun to focus on the development of pharmacological agents that can attenuate the stress response itself, rather than the symptoms associated with stress. Although CRF, which is the main regulator of the stress system, is the focus of current interest, there is an accumulating body of evidence suggesting that the vasopressinergic system may play an equal role in the regulation of the stress response, and that V(1b) receptor antagonists may be of potential therapeutic benefit. The availability of SSR149415, the first selective antagonist for the V(1b) receptor has allowed us to evaluate this hypothesis. SSR149415 is able to attenuate some but not all stress-related behaviors in rodents. While the antidepressant-like activity of the compound was comparable to that of reference antidepressants, the overall profile displayed in anxiety tests was different from that of classical anxiolytics, such as benzodiazepines. The latter were active in a wide range of anxiety models, whereas the V(1b) receptor antagonist showed clear-cut effects only in particularly stressful situations. It is important to note that SSR149415 is devoid of central depressant effects, even at high doses, and does not affect cognitive processes, suggesting a large therapeutic window. Altogether, these findings suggest that V(1b) receptor antagonists might be useful as a treatment for major depression and stress disorders that result from traumatic events. PMID- 12769800 TI - CART peptides as targets for CNS drug development. AB - CART peptides are relatively novel neuropeptides involved in feeding, drug reward and stress. They are formed from a proCART polypeptide that is 89 amino acids in length in the human version. Fragments 42-89 and 49-89 are behaviorally active in feeding and locomotion as well and other functions. These peptides are highly abundant and widely but discretely distributed in the brain, gut, pituitary, adrenals and pancreas. The presence of CART immunoreactivity in specific nuclei of the hypothalamus has led to an examination of icv-injected CART peptides effects on feeding, which have proven to be significantly anorectic. Studies of transgenic animals and humans have also demonstrated a linkage to both obesity and anorexia. Similarly, the localization of CART to sub-regions of the mesolimbic dopamine system has led to demonstration of the effects of CART peptides on locomotor activity and conditioned place preference when injected into the ventral tegmental area (VTA), which are psychostimulant-like in quality. These findings also suggest that CART has the capacity to modulate mesolimbic dopamine, which could have implications for the treatment not only of psychostimulant abuse but also for the treatment of other disorders with mesolimbic dopamine involvement, such as schizophrenia. Other lines of evidence also show that CART peptides are involved in fear and startle behaviors which may have implications for understanding anxiety and stress. An important part of the development of CART mimetics and related drugs would be the identification of CART receptors. At the present time such receptors have not been identified, and much effort should be directed at this problem. Nonetheless, CART peptides offer interesting targets for new drug development for obesity and, potentially, a number of other disorders. PMID- 12769801 TI - Pharmacological findings contribute to the understanding of the main physiological mechanisms of memory retrieval. AB - Recent pharmacological findings have shown that retrieval of one-trial avoidance learning requires glutamate receptors, cAMP-dependent protein kinase and mitogen activated protein kinases in the hippocampus, entorhinal, posterior parietal and anterior cingulate cortex. It requires AMPA but not other type of glutamate receptors or the protein kinases in the amygdala. Retrieval is modulated by dopamine D1, beta-noradrenergic, serotonin 1A and cholinergic receptors in the four cortical structures mentioned, and by beta-noradrenergic receptors in the basolateral amygdala. Further, retrieval is also modulated by peripheral ACTH, glucocorticoids, vasopressin, beta-endorphin and catecholamines; these hormones probably act through beta-noradrenergic receptor systems in the basolateral amygdala. Exposure to novelty or the systemic administration of antidepressant drugs prior to retention tests enhances retrieval, even for very remote memories. The effect of novelty is mediated by molecular mechanisms similar to those of retrieval itself. PMID- 12769802 TI - Biochemical and therapeutic effects of antioxidants in the treatment of Alzheimer's disease, Parkinson's disease, and amyotrophic lateral sclerosis. AB - Aging is a major risk factor for neurodegenerative diseases including Alzheimer's disease (AD), Parkinson's disease (PD), and amyotrophic lateral sclerosis (ALS). An unbalanced overproduction of reactive oxygen species (ROS) may give rise to oxidative stress which can induce neuronal damage, ultimately leading to neuronal death by apoptosis or necrosis. A large body of evidence indicates that oxidative stress is involved in the pathogenesis of AD, PD, and ALS. Several studies have shown that nutritional antioxidants (especially vitamin E and polyphenols) can block neuronal death in vitro, and may have therapeutic properties in animal models of neurodegenerative diseases including AD, PD, and ALS. Moreover, clinical data suggest that nutritional antioxidants might exert some protective effect against AD, PD, and ALS. In this paper, the biochemical mechanisms by which nutritional antioxidants can reduce or block neuronal death occurring in neurodegenerative disorders are reviewed. Particular emphasis will be given to the role played by the nuclear transcription factor -kB (NF-kB) in apoptosis, and in the pathogenesis of neurodegenerative disorders, such as AD, PD, and ALS. The effects of ROS and antioxidants on NF-kB function and their relevance in the pathophysiology of neurodegenerative diseases will also be examined. PMID- 12769803 TI - Emerging signalling and protein interactions mediated via metabotropic glutamate receptors. AB - Metabotropic glutamate receptors (mGlu) are GTP-binding (G) protein-coupled receptors (GPCRs) that are involved in learning and memory, cardiovascular control and motor function. Their structure and pharmacology has been reviewed recently in Current Drug Targets: CNS and Neurological Disorders (Vol. 1, Issue 3) where their roles in a variety of neurological disorders were highlighted. The present review focuses on the emerging evidence for interactions of mGlu receptors with other GPCRs in the CNS at the membrane interface and amongst signaling cascades in the cytosol (e.g. intracellular Ca(2+), cAMP and scaffolding proteins). While initially non-selective activity was thought to be responsible for many atypical responses, increasing evidence points to GPCR interactions in neurons and glia, with adrenoceptors, adenosine receptors, dopamine receptors and muscarinic receptors. For example, group II mGlu receptors were found to be required for group I mGlu receptor induction of long-term potentiation at the postsynaptic terminal. Increasing evidence demonstrates the intimate interaction of adenosine receptors and mGlu receptors, particularly in the regulation of neurotransmitter release. While adenosine itself can be released from astrocytes by co-activation of group II mGlu and beta-adrenergic receptors. Given the complexity of neurological disorders such as ischemic stroke, Alzheimer's disease and epilepsy, exploitation mGlu receptor-associated GPCR interactions may prove efficacious in the treatment of such disorders. PMID- 12769804 TI - Mechanism of action of volatile anesthetics: involvement of intracellular calcium signaling. AB - There have been extensive efforts to characterize the mechanism of action of volatile anesthetics, but their molecular and cellular actions are still a matter of debate. Volatile anesthetics act primarily on synaptic transmission in the central nervous system but proof of this as the predominant mechanism of action remains elusive. Changes in neurotransmitter release may relate to direct interaction of the anesthetic molecule with an ion channel protein or synaptic protein, but can also be a consequence of alterations in intracellular signaling. Calcium is one of the most important messengers in cells and its intracellular concentration may be modified by several agents including volatile anesthetics. Neuronal excitability is in part determined by calcium availability that is controlled by several mechanisms. Because voltage-gated calcium channels (VGCC) play a key role in controlling Ca2+ entry and in initiating cellular responses to stimulation through an elevation of intracellular calcium concentration ([Ca2+](i)), they are thought to be one of the targets for volatile anesthetics. However, [Ca2+](i) can also be altered without the participation of VGCC through receptor-mediated pathways. Indeed, calcium homeostasis is also controlled by plasma membrane Ca2+ -adenosine triphosphatase, sarcoplasmic-endoplasmic reticular Ca2+ -ATPase, the Na+ -Ca2+ exchanger, and mitochondrial Ca2+ sequestration. Alteration of any of those mechanisms that control [Ca2+](i) may lead to a change in presynaptic transmission or postsynaptic excitability. Here we will review some of the recent progress in identifying putative actions of volatile anesthetics, specifically the effect on intracellular calcium homeostasis in neurons. PMID- 12769805 TI - Use of diffusion- and perfusion-weighted magnetic resonance imaging in drug development for ischemic stroke. AB - Diffusion- and perfusion-weighted magnetic resonance imaging (DWI and PWI, respectively) are novel imaging modalities that can detect brain ischemia early in its full extent, can be performed in minutes, can be repeated easily, and allow for follow-up of the ischemic lesion size over time with good spatial and temporal resolution. We have used DWI and PWI in evaluating novel therapeutic approaches for ischemic stroke in numerous studies in the rat and lately in humans. It is now clear that DWI and PWI offer a good combination for safe and reliable evaluation of novel drugs on the size and tissue characteristics of brain ischemia. After inducing focal brain ischemia in the rat, one can first detect the presence and extent of ischemia by DWI and hypoperfusion by PWI, calculate the volume of ischemic brain tissue, and then follow the development of the ischemic lesion over time for several hours during treatment, thus detecting in vivo effects of the novel drug on brain ischemia. Successful reperfusion (either mechanically or as a result of thrombolytic therapy) can also be detected easily. DWI and PWI when performed before starting treatment can also exclude the pretreatment bias, a potential reason for false-positive studies in which proper imaging studies are not employed. Thus we can determine the in vivo efficacy (or lack of efficacy) of new therapeutic regimens (both neuroprotective and thrombolytic) rapidly, safely, and reliably by using a small sample size only, and adapt the same strategy to clinical trials. PMID- 12769806 TI - 3,4-dihydroxyphenylacetaldehyde: a potential target for neuroprotective therapy in Parkinson's disease. AB - The simplest explanation for the selective loss of substantia nigra (SN) dopamine (DA) neurons in Parkinson's disease (PD) is that DA or a metabolite is neurotoxic. Recently, a series of investigations implicate the MAO metabolite of DA, 3,4-dihydroxyphenylacetaldehyde (DOPAL), as the critical endogenous toxin which triggers DA neuron loss in PD: 1. Hereditary PD contains mutations in the gene for alpha-synuclein (alpha-syn). Investigations implicate a DA metabolite as mediator of alpha-syn neurotoxicity, and DOPAL is 1000-fold more toxic than DA in vivo. 2. A deficit in mitochondrial complex I is found in PD SN. Inhibition of complex I causes increases in DOPAL levels and death of DA neurons in vitro and in vivo. 3. L-DOPA, the precursor of DA, which is used to treat PD, is toxic and contributes to the progression of PD. L-DOPA-treated rats have an 18-fold increase in striatal DOPAL. 4. Free hydroxyl radicals (.OH) trigger aggregation of alpha-syn to its toxic form. DOPAL with H(2)O(2) generates.OH radicals. These investigations provide several therapeutic strategies to limit DOPAL toxicity and progression of PD: 1. Delaying the start of L-DOPA therapy by early use of DA receptor agonists, which may also be free radical scavengers, limits the amount of DOPAL formed from L-DOPA. 2. Nonspecific MAO inhibitors may more effectively decrease production of DOPAL from DA than MAO-B inhibitors. 3. Newer more potent and targeted free radical scavengers could block DOPAL toxicity. 4. Coenzyme Q(10) increases complex I activity and nicotine adenine dinucleotide (NAD) synthesis, and thereby could enhance DOPAL catabolism by aldehyde dehydrogenase, which uses NAD as a cofactor. 5. DA uptake blockers could be used to limit intraneuronal DOPAL production. 6. Tauroursodeoxycholic acid, an inhibitor of apoptosis shown to be effective in models of Huntington's disease, may also prove effective in blocking DOPAL toxicity in PD. 7. Agents which block aggregation of alpha-syn should limit DOPAL toxicity. PMID- 12769807 TI - Gastrointestinal disturbances in anorexia nervosa and bulimia nervosa. AB - Disturbances in the functioning of the upper gastrointestinal (GI ) tract have been described in both Anorexia Nervosa (AN) and Bulimia Nervosa (BN). Patients with AN experience substantial delays in gastric emptying as well as constipation. These problems may give rise to significant medical complications and may contribute to increased difficulties with refeeding and weight restoration. Reported GI disturbances in BN include increased gastric capacity, diminished gastric relaxation, delayed gastric emptying, diminished release of cholecystokinin (CCK) and abnormalities of enteric autonomic function, all of which may play a role in perpetuation of the syndrome. This article reviews evidence for the most common disturbances of GI function in AN and BN and discusses potential GI targets for therapeutic intervention. PMID- 12769808 TI - Osteoporosis in anorexia nervosa: a brittle future? AB - Anorexia nervosa commonly occurs during adolescence, a critical period for the acquisition of peak bone mass. Osteopenia is an early and serious complication associated with anorexia nervosa. Whether the osteopenia observed in adolescents with anorexia nervosa is reversible is unknown. The possible mechanisms for the osteopenia observed in adolescents with anorexia nervosa are complex and poorly understood. The purpose of this paper is to review a number of the identified factors that influence the attainment of peak bone mass in anorexia nervosa and to examine the best treatment options for optimizing skeletal mineralization in adolescents with anorexia nervosa. PMID- 12769809 TI - Drug therapy for patients with eating disorders. AB - The purpose of this article is to review the psychopharmacology treatment literature for patients with eating disorders including bulimia nervosa, anorexia nervosa and binge eating disorder. The best-developed treatment literature concerns bulimia nervosa, which has been studied now in several dozen pharmacological treatment studies. The agents most commonly used are the antidepressants, with particular focus on the selective serotonin reuptake inhibitors including fluoxetine hydrochloride. These agents clearly impact significantly on the frequency of abnormal eating behaviors such as binge eating and purging. However, subjects treated with these drugs rarely achieve remission. Pharmacotherapy of anorexia nervosa has also traditionally focused on the use of antidepressants and there is some evidence that the use of SSRIs may help in preventing relapse in weight restored patients. Recently interest has developed in the use of atypical neuroleptics to help with the obsessionality and resistance to treatment frequently seen in low weight patients, the most commonly employed agent being olanzapine. Pharmacotherapy of binge-eating disorder is now being intensively investigated. In general medication alone seems inferior to psychotherapy in the short term. Antidepressants can increase the amount of weight loss when combined with psychological treatment and also appear to benefit symptoms such as depression. Further data are needed, but a number of drugs appear promising. PMID- 12769810 TI - Candidate genes in eating disorders. AB - Environmental influences, as well as biological and genetic factors influence risk for eating disorders. Family and twin studies have shown that eating disorders are familial and suggest the influence of genetic factors on their etiology. Positive associations have been observed for some candidate genes that have been studied (such as 5HT2A receptor gene); however, the field has been plagued by nonreplications. In this paper we review the extant association studies of eating disorders. PMID- 12769811 TI - Candidate gene analysis of the Price Foundation anorexia nervosa affected relative pair dataset. AB - The eating disorders are severe psychiatric illnesses with significant morbidity and mortality that exhibit statistically significant familial risk and heritability, providing support for a molecular genetic approach toward defining etiological factors. An emerging candidate gene literature has concentrated on serotinergic and dopaminergic candidates. With the financial support of the Price Foundation, a group of investigators initiated an international multi-center collaboration (Price Foundation Collaborative Group) in 1995 to study the genetics of anorexia and bulimia nervosa by collecting and analyzing phenotypes and genotypes of individuals and their relatives affected with eating disorders. The first sample of families collected by this collaborative group, known as the Price Foundation Anorexia Nervosa Affected Relative Pair (AN-ARP) dataset, was ascertained on an proband affected with Anorexia Nervosa (AN), with relative pairs affected with the eating disorders AN, Bulimia Nervosa or Eating Disorders Not Otherwise Specified [1]. Biognosis U.S., Inc. was founded to identify and characterize candidate susceptibility genes for anorexia and bulimia nervosa phenotypes in the Price Foundation eating disorder datasets. During 2000-2001, Biognosis U.S., Inc. developed and implemented a research program with a focus on the analysis of candidate genes nominated by neurochemical characteristics of eating disorder patients [2], serotonergic and dopaminergic candidate gene polymorphisms [3], neuroendocrine regulation of appetite [4], and by a positional hypothesis from a linkage analysis of the AN-ARP dataset [5]. This report reviews the anorexia nervosa candidate gene literature through 2001, the candidate gene research program implemented at Biognosis U.S., Inc. and selected candidate gene findings in the AN-ARP dataset derived from that research program. PMID- 12769812 TI - A review of neuropeptide and neuroendocrine dysregulation in anorexia and bulimia nervosa. AB - Neuropeptides play an important role in the regulation of feeding behavior and obesity. The mechanisms for controlling food intake involve a complicated interplay between peripheral systems (including gustatory stimulation, gastrointestinal peptide secretion, and vagal afferent nerve responses) and central nervous system (CNS) neuropeptides and/or monoamines. These neuronal systems include neuropeptides (CRH, opioids, neuropeptide-Y (NPY) and peptide YY (PYY), vasopressin and oxytocin, CCK, and leptin) and monamines (serotonin, dopamine, norepinephrine). In addition to regulating eating behavior, a number of CNS neuropeptides participate in the regulation of neuroendocrine pathways. Thus, clinical studies have evaluated the possibility that CNS neuropeptide alterations may contribute to dysregulated secretion of the gonadal hormones, cortisol, thyroid hormones and growth hormone in the eating disorders. Most of the neuroendocrine and neuropeptide alterations apparent during symptomatic episodes of AN and BN tend to normalize after recovery. This observation suggests that most of the disturbances are consequences rather than causes of malnutrition, weight loss and/or altered meal patterns. Still, an understanding of these neuropeptide disturbances may shed light on why many people with AN or BN cannot easily "reverse" their illness and even after weight gain and normalized eating patterns, many individuals who have recovered from AN or BN have physiological, behavioral and psychological symptoms that persist for extended periods of time. PMID- 12769813 TI - Neurotransmitter and imaging studies in anorexia nervosa: new targets for treatment. AB - Anorexia and Bulimia Nervosa are disorders of unknown etiology that invariably begin during adolescence and near in time to puberty in young women. These disorders are associated with aberrant eating behaviors, body image distortions, impulse and mood disturbances, as well as characteristic temperament and personality traits. It is well known that malnutrition produces changes in neuroendocrine function. More recently, disturbances in neuronal systems have been found to play a role in the modulation of feeding, mood, and impulse control. These neuronal systems include neuropeptides (CRH, opioids, neuropeptide Y (NPY) and peptide YY (PYY), vasopressin and oxytocin, CCK, and leptin) and monoamines (serotonin, dopamine, norepinephrine). Disturbances of most of these neuronal systems have been found when people are ill with an eating disorder, but it was not certain whether they were a cause or consequence of symptoms. In order to address these questions, a growing number of studies have investigated whether neuromodulatory disturbances persist after recovery. Studies from several centers tend to show altered serotonin activity persists after prolonged normalization of weight, nutrition, and menstrual function, as do anxiety, obsessionality, and perfectionism. While there are fewer data, there may be persistent alterations of dopamine or some neuropeptides in some subjects in a recovered state. The inaccessibility of the central nervous system has made it difficult to understand brain and behavior. In the past decade, new tools, such as brain imaging, have offered the possibility of better characterization of complex neuronal function and behavior. Such studies have tended to consistently find that alterations of brain regions, such as the temporal lobe, occur in people who are ill with anorexia nervosa and appear to persist after some degree of weight gain and recovery. New imaging technology, that marries Positron Emission Tomography (PET) imaging with selective neurotransmitter radioligands, confirms that altered serotonin neuronal pathway activity persists after recovery from an eating disorder and supports the possibility that these psychobiological alterations might contribute to traits, such as increased anxiety or extremes of impulse control, that, in turn, may contribute to a vulnerability to the development of an eating disorder. In summary, studies of pathophysiology are starting to nominate new candidates for treatment leading to the possibility of finding effective treatments for this often chronic or fatal disorder. PMID- 12769814 TI - Effect of maternal obstructive cholestasis during pregnancy on the biliary transport of horseradish peroxidase in the rat offspring. AB - MOCP (maternal obstructive cholestasis during pregnancy) induces a reversible impairment in bile formation in young rats born to these mothers. The aim of the present study was to gain information on the effects of MOCP on the maturation of pathways involved in protein secretion into bile in young (4-week-old) rats. The amount of hepatic alpha-tubulin and the structure of the microtubular network were apparently not affected by MOCP. HRP (horseradish peroxidase) was used as a model protein, and its secretion into bile after administration through the jugular vein was measured. In adult (8-week-old) rats, two peaks of HRP output into bile were observed following administration: an early peak presumably due to paracellular transfer, and a late peak presumably due to transcytosis. In young rats (4 weeks old), the early peak was similar to that of adult animals, and was not affected by MOCP. However, the late peak was markedly smaller in young control rats, and was further reduced by MOCP. Brefeldin A decreased, whereas taurocholate did not change, the early peak, whereas both affected the transcytotic transport of HRP. Brefeldin A delayed HRP secretion (similarly in control and MOCP groups), without affecting cumulative output, whereas taurocholate accelerated the transcytotic transport of HRP in the control group, but not in the MOCP group. These results suggest that MOCP affects the maturation of hepatocyte mechanisms involved in the transcytotic secretion of HRP into bile. PMID- 12769815 TI - Labour-associated changes in adrenomedullin content in human placenta and fetal membranes. AB - The aim of the present study was to determine the effects of labour and mode of delivery on human placental and fetal membrane content of adrenomedullin (AdM). Placentas and fetal membranes were collected either at term or pre-term gestation from women either in labour or not in labour, and AdM was measured in tissue extracts by specific RIA. There were significant increases in AdM concentrations in amnion and choriodecidua for the in-labour group compared with the not-in labour group for both pre-term and term gestations. There was no difference in AdM concentration in placental tissue between labour groups. This study provides evidence that fetal membrane AdM is increased in amniotic and choriodecidual tissues in response to labour, and suggests that it may play a role during human labour. PMID- 12769816 TI - Membrane disruption and cytotoxicity of hydrophobic N-alkylated imino sugars is independent of the inhibition of protein and lipid glycosylation. AB - The N-alkyl moiety of N-alkylated imino sugars is crucial for therapeutic activities of these compounds as inhibitors of glycosphingolipid (GSL) biosynthesis and as antivirals. The improved potency afforded by a long N-alkyl moiety is coincident with increased compound-induced cytotoxicity. Therefore, in the present study, we examined the mechanism of this cytotoxicity in detail. Despite N-butyl-deoxynojirimycin and N-butyl-deoxygalactonojirimycin inhibiting the glycosylation of ceramide to glucosylceramide, ceramide levels did not increase in HL60 cells treated with these compounds. Long-chain N-alkylated imino sugars were toxic to cells at concentrations considerably lower than the critical micellar concentrations for these compounds and consequently did not solubilize radioactively labelled cellular proteins and lipids. However, membrane disruption and cell fragmentation did increase in a concentration- and chain-length dependent manner. These results are consistent with previously proposed interactions between surface-active amphiphiles and protein-containing lipid membranes when drug concentrations are below the critical micellar concentration. Taken together, these results demonstrate that the cellular toxicity of hydrophobic N-alkylated imino sugars is due to cell lysis and cell fragmentation and, most importantly, is not related to the beneficial therapeutic effects of these compounds on protein and in lipid glycosylation. This information will aid in the future development of more selective imino sugar therapeutics for the treatment of human disease. PMID- 12769817 TI - Replacement of a cytosolic copper/zinc superoxide dismutase by a novel cytosolic manganese superoxide dismutase in crustaceans that use copper (haemocyanin) for oxygen transport. AB - The blue crab, Callinectes sapidus, which uses the copper-dependent protein haemocyanin for oxygen transport, lacks the ubiquitous cytosolic copper-dependent enzyme copper/zinc superoxide dismutase (Cu,ZnSOD) as evidenced by undetectable levels of Cu,ZnSOD activity, protein and mRNA in the hepatopancreas (the site of haemocyanin synthesis) and gills. Instead, the crab has an unusual cytosolic manganese SOD (cytMnSOD), which is retained in the cytosol, because it lacks a mitochondrial transit peptide. A second familiar MnSOD is present in the mitochondria (mtMnSOD). This unique phenomenon occurs in all Crustacea that use haemocyanin for oxygen transport. Molecular phylogeny analysis suggests the MnSOD gene duplication is as old as the origin of the arthropod phylum. cytMnSOD activity in the hepatopancreas changes during the moulting cycle of the crab. Activity is high in intermoult crabs and non-detectable in postmoult papershell crabs. mtMnSOD is present in all stages of the moulting cycle. Despite the lack of cytCu,ZnSOD, crabs have an extracellular Cu,ZnSOD (ecCu,ZnSOD) that is produced by haemocytes, and is part of a large, approx. 160 kDa, covalently linked protein complex. ecCu,ZnSOD is absent from the hepatopancreas of intermoult crabs, but appears in this tissue at premoult. However, no ecCu,ZnSOD mRNA can be detected, suggesting that the protein is recruited from the haemolymph. Screening of different taxa of the arthropod phylum for Cu,ZnSOD activity shows that those crustaceans that use haemoglobin for oxygen transport have retained cytCu,ZnSOD. It appears, therefore, that the replacement of cytCu,ZnSOD with cytMnSOD is part of an adaptive response to the dynamic, haemocyanin-linked, fluctuations in copper metabolism that occur during the moulting cycle of the crab. PMID- 12769818 TI - Cobalamin (vitamin B12)-deficiency-induced changes in the proteome of rat cerebrospinal fluid. AB - We studied the changes in the proteome of CSF (cerebrospinal fluid) in two animal models of Cbl (cobalamin) deficiency: TGX (totally gastrectomized) rats and rats fed a Cbl-D (Cbl-deficient) diet. Two-dimensional PAGE was used to detect qualitative and quantitative variations in proteins in the CSF samples. The peak increase in total CSF protein concentration was observed 4 months after TG (total gastrectomy) and after 6 months of eating a Cbl-D diet. There is a specific increase 4 months after TG in the spots corresponding to alpha1-antitrypsin and the de novo presence of thiostatin and haptoglobin beta. Cbl-replacement treatment in 4-month-TGX rats corrected these alterations in the CSF proteome. However, most of the CSF proteome alterations attenuated in Cbl-untreated 8-month TGX rats and in rats fed a Cbl-D diet for 16 months. Transthyretin concentration varied slightly in the CSF of both types of Cbl-D rat, whereas the relative abundance of prostaglandin D synthase rose sharply in the CSF of the rats fed a Cbl-D diet for 16 months. We have demonstrated previously that the histological and ultrastructural CNS (central nervous system) damage in both types of Cbl-D rat appears within 2-3 months of Cbl deficiency, and thus appears to precede the alterations in the CSF proteome. The CSF proteome patterns of rats in which phlogosis was induced in or outside the CNS are quite different from those of the CSF of Cbl-D rats. All these findings demonstrate that the alterations in the CSF proteome of Cbl-D rats are specifically linked to Cbl deficiency. PMID- 12769819 TI - Modulation of DNA topoisomerase II alpha promoter activity by members of the Sp (specificity protein) and NF-Y (nuclear factor Y) families of transcription factors. AB - Topo IIalpha (topoisomerase IIalpha) is a major target of several commonly used anticancer drugs and is subject to down-regulation at the transcriptional level in some drug-resistant cell lines and tumours in response to chemotherapy. Clinical resistance to such drugs has been correlated with down-regulation of topo IIalpha at transcription in some drug-resistant cell lines and tumours. Putative binding sites for a variety of transcription factors, including Sp1 (specificity protein 1) and NF-Y (nuclear factor Y) have previously been identified in the topo IIalpha promoter, but their functional significance and interactions have not been described following exposure to anti-cancer drugs. The binding of these factors to specific putative regulatory elements in the topo IIalpha promoter was studied using electrophoretic-mobility-shift assays. Sp1 was found to bind strongly to both distal and proximal GC-rich elements and NF-Y to ICB1 (the first inverted CCAAT box). The functional significance of transcription factor binding was studied using transient transfection of HeLa cells using a luciferase reporter driven by a 617-bp minimal promoter containing point mutations in putative regulatory elements. Sp1 and NF-Y were both found to be transcriptional modulators with activator or repressor functions depending on protein/DNA context. Moreover, a functional interaction between Sp1 and NF-Y bound at proximal elements was observed. PMID- 12769820 TI - A simple dependence between protein evolution rate and the number of protein protein interactions. AB - BACKGROUND: It has been shown for an evolutionarily distant genomic comparison that the number of protein-protein interactions a protein has correlates negatively with their rates of evolution. However, the generality of this observation has recently been challenged. Here we examine the problem using protein-protein interaction data from the yeast Saccharomyces cerevisiae and genome sequences from two other yeast species. RESULTS: In contrast to a previous study that used an incomplete set of protein-protein interactions, we observed a highly significant correlation between number of interactions and evolutionary distance to either Candida albicans or Schizosaccharomyces pombe. This study differs from the previous one in that it includes all known protein interactions from S. cerevisiae, and a larger set of protein evolutionary rates. In both evolutionary comparisons, a simple monotonic relationship was found across the entire range of the number of protein-protein interactions. In agreement with our earlier findings, this relationship cannot be explained by the fact that proteins with many interactions tend to be important to yeast. The generality of these correlations in other kingdoms of life unfortunately cannot be addressed at this time, due to the incompleteness of protein-protein interaction data from organisms other than S. cerevisiae. CONCLUSIONS: Protein-protein interactions tend to slow the rate at which proteins evolve. This may be due to structural constraints that must be met to maintain interactions, but more work is needed to definitively establish the mechanism(s) behind the correlations we have observed. PMID- 12769821 TI - Identification of signature and primers specific to genus Pseudomonas using mismatched patterns of 16S rDNA sequences. AB - BACKGROUND: Pseudomonas, a soil bacterium, has been observed as a dominant genus that survives in different habitats with wide hostile conditions. We had a basic assumption that the species level variation in 16S rDNA sequences of a bacterial genus is mainly due to substitutions rather than insertion or deletion of bases. Keeping this in view, the aim was to identify a region of 16S rDNA sequence and within that focus on substitution prone stretches indicating species level variation and to derive patterns from these stretches that are specific to the genus. RESULTS: Repeating elements that are highly conserved across different species of Pseudomonas were considered as guiding markers to locate a region within the 16S gene. Four repeating patterns showing more than 80% consistency across fifty different species of Pseudomonas were identified. The sub-sequences between the repeating patterns yielded a continuous region of 495 bases. The sub sequences after alignment and using Shanon's entropy measure yielded a consensus pattern. A stretch of 24 base positions in this region, showing maximum variations across the sampled sequences was focused for possible genus specific patterns. Nine patterns in this stretch showed nearly 70% specificity to the target genus. These patterns were further used to obtain a signature that is highly specific to Pseudomonas. The signature region was used to design PCR primers, which yielded a PCR product of 150 bp whose specificity was validated through a sample experiment. CONCLUSIONS: The developed approach was successfully applied to genus Pseudomonas. It could be tried in other bacterial genera to obtain respective signature patterns and thereby PCR primers, for their rapid tracking in the environmental samples. PMID- 12769822 TI - MALDI/MS peptide mass fingerprinting for proteome analysis: identification of hydrophobic proteins attached to eucaryote keratinocyte cytoplasmic membrane using different matrices in concert. AB - BACKGROUND: MALDI-TOF-MS has become an important analytical tool in the identification of proteins and evaluation of their role in biological processes. A typical protocol consists of sample purification, separation of proteins by 2D PAGE, enzymatic digestion and identification of proteins by peptide mass fingerprint. Unfortunately, this approach is not appropriate for the identification of membrane or low or high pI proteins. An alternative technique uses 1D-PAGE, which results in a mixture of proteins in each gel band. The direct analysis of the proteolytic digestion of this mixture is often problematic because of poor peptide detection and consequent poor sequence coverage in databases. Sequence coverage can be improved through the combination of several matrices. RESULTS: The aim of this study was to trust the MALDI analysis of complex biological samples, in order to identify proteins that interact with the membrane network of keratinocytes. Peptides obtained from protein trypsin digestions may have either hydrophobic or hydrophilic sections, in which case, the direct analysis of such a mixture by MALDI does not allow desorbing of all peptides. In this work, MALDI/MS experiments were thus performed using four different matrices in concert. The data were analysed with three algorithms in order to test each of them. We observed that the use of at least two matrices in concert leads to a twofold increase of the coverage of each protein. Considering data obtained in this study, we recommend the use of HCCA in concert with the SA matrix in order to obtain a good coverage of hydrophilic proteins, and DHB in concert with the SA matrix to obtain a good coverage of hydrophobic proteins. CONCLUSION: In this work, experiments were performed directly on complex biological samples, in order to see systematic comparison between different matrices for real-life samples and to show a correlation that will be applicable to similar studies. When 1D gel is needed, each band may contain a great number of proteins, each present in small amounts. To improve the proteins coverage, we have performed experiments with some matrices in concert. These experiments enabled reliable identification of proteins, without the use of Nanospray MS/MS experiments. PMID- 12769824 TI - The development of Trypanosoma brucei within the tsetse fly midgut observed using green fluorescent trypanosomes. AB - BACKGROUND: The protozoan pathogen, Trypanosoma brucei, undergoes complex cycles of differentiation and multiplication in its vector, the tsetse fly, genus Glossina. Flies are refractory to infection and resistance mechanisms operate at a number of levels and timepoints. Here we have used highly conspicuous green fluorescent trypanosomes to study the early events in establishment of infection in the fly midgut. RESULTS: Less than 10% of the bloodstream form trypanosomes in the infected feed differentiated into viable procyclics. Up to day 3, trypanosomes were found in the bloodmeal in every fly examined, and increased in number between days 1 and 3. Flies dissected on days 5 and 6 fell into 2 clearly distinct groups: those with high numbers of trypanosomes and those with undetectable infection. Trypanosomes were found in the ectoperitrophic space and proventriculus from 6 days following the infective feed. CONCLUSION: Trypanosomes that have undergone successful differentiation appear to experience an environment within the midgut suited to their unrestricted growth for the first 3 days. After this time, a process of attrition is evident in some flies, which leads to the complete elimination of infection. By day 5, flies fall into 2 groups according to the level of infection: high or undetectable. This timecourse coincides with lectin secretion, development of the PM and the digestion and movement of the bloodmeal along the gut. Further experiments are needed to discriminate between these factors. PMID- 12769825 TI - Impact of ivermectin on onchocerciasis transmission: assessing the empirical evidence that repeated ivermectin mass treatments may lead to elimination/eradication in West-Africa. AB - BACKGROUND: The Onchocerciasis Control Program (OCP) in West Africa has been closed down at the end of 2002. All subsequent control will be transferred to the participating countries and will almost entirely be based on periodic mass treatment with ivermectin. This makes the question whether elimination of infection or eradication of onchocerciasis can be achieved using this strategy of critical importance. This study was undertaken to explore this issue. METHODS: An empirical approach was adopted in which a comprehensive analysis was undertaken of available data on the impact of more than a decade of ivermectin treatment on onchocerciasis infection and transmission. Relevant entomological and epidemiological data from 14 river basins in the OCP and one basin in Cameroon were reviewed. Areas were distinguished by frequency of treatment (6-monthly or annually), endemicity level and additional control measures such as vector control. Assessment of results were in terms of epidemiological and entomological parameters, and as a measure of inputs, therapeutic and geographical coverage rates were used. RESULTS: In all of the river basins studied, ivermectin treatment sharply reduced prevalence and intensity of infection. Significant transmission, however, is still ongoing in some basins after 10-12 years of ivermectin treatment. In other basins, transmission may have been interrupted, but this needs to be confirmed by in-depth evaluations. In one mesoendemic basin, where 20 rounds of four-monthly treatment reduced prevalence of infection to levels as low as 2-3%, there was significant recrudescence of infection within a few years after interruption of treatment. CONCLUSIONS: Ivermectin treatment has been very successful in eliminating onchocerciasis as a public health problem. However, the results presented in this paper make it almost certain that repeated ivermectin mass treatment will not lead to the elimination of transmission of onchocerciasis from West Africa. Data on 6-monthly treatments are not sufficient to draw definitive conclusions. PMID- 12769823 TI - Animal models of copper-associated liver disease. AB - Recent advances in molecular biology have made possible the identification of genetic defects responsible for Wilson's disease, Indian childhood cirrhosis and copper toxicosis in Long Evans Cinnamon rats, toxic milk mice, and Bedlington terriers. The Wilson's disease gene is localized on human chromosome 13 and codes for ATP7B, a copper transporting P-type ATPase. A genetic defect similar to that of Wilson's disease occurs in Long Evans Cinnamon rats and toxic milk mice. Familial copper storage disorders in Bedlington and West Highland white terriers are associated with early subclinical disease, and copper accumulation with subsequent liver injury culminating in cirrhosis. The canine copper toxicosis locus in Bedlington terriers has been mapped to canine chromosome region CFA 10q26. Recently, a mutated MURR1 gene was discovered in Bedlington terriers affected with the disease. Idiopathic childhood cirrhosis is biochemically similar to copper toxicosis in Bedlington terriers, but clinically much more severe. Both conditions are characterized by the absence of neurologic damage and Kayser-Fleisher rings, and normal ceruloplasmin levels. A recent study added North Ronaldsay sheep to the list of promising animal models to study Indian childhood cirrhosis. Morphologic similarities between the two conditions include periportal to panlobular copper retention and liver changes varying from active hepatitis to panlobular pericellular fibrosis, and cirrhosis. Certain copper associated disorders, such as chronic active hepatitis in Doberman pinschers and Skye terrier hepatitis are characterized by copper retention secondary to the underlying disease, thus resembling primary biliary cirrhosis in humans. Copper associated liver disease has increasingly being recognized in Dalmatians. Copper associated liver diseases in Dalmatians and Long Evans Cinnamom rats share many morphologic features. Fulminant hepatic failure in Dalmatians is characterized by high serum activities of alanine aminotransferase and aspartate aminotransferase, and severe necrosis of centrilobular areas (periacinar, zone 3) hepatocytes. Macrophages and surviving hepatocytes contain copper-positive material. Liver disease associated with periacinar copper accumulation has also been described in Siamese cats. Many questions regarding copper metabolism in mammals, genetic background, pathogenesis and treatment of copper-associated liver diseases remain to be answered. This review describes the similarities between the clinico pathological features of spontaneous copper-associated diseases in humans and domestic animals. PMID- 12769826 TI - The tissue microarray data exchange specification: a community-based, open source tool for sharing tissue microarray data. AB - BACKGROUND: Tissue Microarrays (TMAs) allow researchers to examine hundreds of small tissue samples on a single glass slide. The information held in a single TMA slide may easily involve Gigabytes of data. To benefit from TMA technology, the scientific community needs an open source TMA data exchange specification that will convey all of the data in a TMA experiment in a format that is understandable to both humans and computers. A data exchange specification for TMAs allows researchers to submit their data to journals and to public data repositories and to share or merge data from different laboratories. In May 2001, the Association of Pathology Informatics (API) hosted the first in a series of four workshops, co-sponsored by the National Cancer Institute, to develop an open, community-supported TMA data exchange specification. METHODS: A draft tissue microarray data exchange specification was developed through workshop meetings. The first workshop confirmed community support for the effort and urged the creation of an open XML-based specification. This was to evolve in steps with approval for each step coming from the stakeholders in the user community during open workshops. By the fourth workshop, held October, 2002, a set of Common Data Elements (CDEs) was established as well as a basic strategy for organizing TMA data in self-describing XML documents. RESULTS: The TMA data exchange specification is a well-formed XML document with four required sections: 1) Header, containing the specification Dublin Core identifiers, 2) Block, describing the paraffin-embedded array of tissues, 3)Slide, describing the glass slides produced from the Block, and 4) Core, containing all data related to the individual tissue samples contained in the array. Eighty CDEs, conforming to the ISO-11179 specification for data elements constitute XML tags used in the TMA data exchange specification. A set of six simple semantic rules describe the complete data exchange specification. Anyone using the data exchange specification can validate their TMA files using a software implementation written in Perl and distributed as a supplemental file with this publication. CONCLUSION: The TMA data exchange specification is now available in a draft form with community-approved Common Data Elements and a community-approved general file format and data structure. The specification can be freely used by the scientific community. Efforts sponsored by the Association for Pathology Informatics to refine the draft TMA data exchange specification are expected to continue for at least two more years. The interested public is invited to participate in these open efforts. Information on future workshops will be posted at http://www.pathologyinformatics.org (API we site). PMID- 12769827 TI - Factors related to the practice of breast self examination (BSE) and Pap smear screening among Malaysian women workers in selected electronics factories. AB - BACKGROUND: The Malaysian Ministry of Health promotes breast self-examination (BSE) for all women, and Pap smear screening every three years for all sexually active women ages 20 years and above. The objectives of this paper were to examine the practice of these two screening tests among women production workers in electronics factories, and to identify factors related to practice. METHODS: This was a cross-sectional survey of women production workers from ten electronics factories. Data was collected by a self-administered questionnaire from a total of 1,720 women. The chi-square test, odds ratio and binomial logistic regression were used in bivariate and multivariate analysis. RESULTS: Prevalence rates were 24.4% for BSE once a month, and 18.4% for Pap smear examination within the last three years. Women who were significantly more likely to perform BSE every month were 30 years and older, Malays, with upper secondary education and above, answered the BSE question correctly, and had a Pap smear within the last three years. The proportion of women who had a Pap smear within the last three years were significantly higher among those who were older, married, with young children, on the contraceptive pill or intra-uterine device, had a medical examination within the last five years, answered the Pap smear question correctly, and performed BSE monthly. CONCLUSION: Screening practice rates in this study were low when compared to national rates. Socio-demographic and health care factors significantly associated with screening practice are indicative of barriers which should be further understood so that more effective educational and promotional strategies could be developed. PMID- 12769828 TI - Efficiently folding and circularly permuted variants of the Sapphire mutant of GFP. AB - BACKGROUND: The green fluorescent protein (GFP) has been widely used in cell biology as a marker of gene expression, label of cellular structures, fusion tag or as a crucial constituent of genetically encoded biosensors. Mutagenesis of the wildtype gene has yielded a number of improved variants such as EGFP or colour variants suitable for fluorescence resonance energy transfer (FRET). However, folding of some of these mutants is still a problem when targeted to certain organelles or fused to other proteins. RESULTS: By directed rational mutagenesis, we have produced a new variant of the Sapphire mutant of GFP with improved folding properties that turns out to be especially beneficial when expressed within organelles or as a fusion tag. Its absorption spectrum is pH-stable and the pKa of its emission is 4.9, making it very resistant to pH perturbation inside cells. CONCLUSION: "T-Sapphire" and its circular permutations can be used as labels of proteins or cellular structures and as FRET donors in combination with red-fluorescent acceptor proteins such as DsRed, making it possible to completely separate donor and acceptor excitation and emission in intensity-based FRET experiments. PMID- 12769830 TI - Should tumor depth be included in prognostication of soft tissue sarcoma? AB - BACKGROUND: Most staging systems for soft tissue sarcoma are based on histologic malignancy-grade, tumor size and tumor depth. These factors are generally dichotomized, size at 5 cm. We believe it is unlikely that tumor depth per se should influence a tumor's metastatic capability. Therefore we hypothesized that the unfavourable prognostic importance of depth could be explained by the close association between size and depth, deep-seated tumors on average being larger than the superficial ones. When tumor size is dichotomized, this effect should be most pronounced in the large size (>5 cm) group in which the size span is larger. METHODS: We analyzed the associations between tumor size and depth and the prognostic importance of grade, size and depth in a population-based series of 490 adult patients with soft tissue sarcoma of the extremity or trunk wall with complete, 4.5 years minimum, follow-up. RESULTS: Multivariate analysis showed no major prognostic effect of tumor depth when grade and size were taken into account. The mean size of small tumors was the same whether superficial or deep but the mean size of large and deep-seated tumors were one third larger than that of large but superficial tumors. Tumor depth influenced the prognosis in the subset of high-grade and large tumors. In this subset deep-seated tumors had poorer survival rate than superficial tumors, which could be explained by the larger mean size of the deep-seated tumors. CONCLUSION: Most of the prognostic value of tumor depth in soft tissue sarcomas of the extremity or trunk wall can be explained by the association between tumor size and depth. PMID- 12769829 TI - Breastfeeding in Bolivia - information and attitudes. AB - BACKGROUND: The main objective of the present study was to investigate the relationship between the attitudes of the mother and her family towards breastfeeding and the actual feeding pattern in a Bolivian population. A second objective was to study the relationship between breastfeeding information, specified according to source and timing, and feeding pattern. METHODS: Cross sectional interviews with 420-502 Bolivian mothers with an infant less than or equal to 1 year of age. Duration of exclusive breastfeeding, use of prelacteal food and/or colostrum were the main outcome measures. RESULTS: The attitudes of the mother, her partner (the infant's father) and the infant's grandmother towards breastfeeding did not influence the infant feeding pattern. Women who had received breastfeeding information from health care personnel before birth or on the maternity ward breastfed exclusively for a longer duration (adjusted p = 0.0233) and avoided prelacteal food to a greater extent (adjusted odds ratio (AOR) = 0.42; 95% confidence interval for adjusted odds ratio (95% CI AOR) = 0.23 0.72). Information from a doctor before birth or on the maternity ward was associated with less use of prelacteal food (AOR = 0.53; 95% CI AOR = 0.31-0.93), an increased use of colostrum (AOR = 3.30; 95% CI AOR = 1.16-9.37), but was not linked to the duration of exclusive breastfeeding (p = 0.1767). CONCLUSION: The current study indicates that breastfeeding information delivered by health care personnel in a non-trial setting may affect the infant feeding pattern including the use of prelacteal foods and colostrum. There was no evidence that the attitudes of the mother, or the infant's father or grandmother influenced actual feeding behavior. The lack of a "negative or neutral attitude" towards breastfeeding in the participants of the current study does, however, diminish the chances to link attitude to feeding behavior. PMID- 12769831 TI - Seroprevalence of Toxoplasma gondii infection among veterinary staff in Ontario, Canada (2002): implications for teratogenic risk. AB - BACKGROUND: Toxoplasma gondii infection is embryotoxic in humans. It is mainly transmitted through raw/undercooked meat and ingestion of oocysts in cat feces. There remains controversy about the actual risk of cats transmitting the disease to humans. Our primary objective was to determine the seroprevalence of T. gondii antibody among veterinary staff, to ascertain whether they have an increased risk through occupational exposure. Our secondary objective was to examine their practices regarding cats, toxoplasma infection, and pregnancy. METHODS: Veterinary staff attending the 2002 Annual Ontario Veterinary Medical Association Conference were invited to discuss their toxoplasma seroprevalence. Interested attendees completed a questionnaire and a physician drew blood samples to determine T. gondii titres using the ELISA IgG test. RESULTS: We collected 161 completed questionnaires, and 141 blood samples. There were 20 (14.2%, CI 95%:8.4 19.9%) reactive titres among the veterinarian staff (80% females aged 30-45). All were regularly exposed to cats, washed their hands when in contact and few wore gloves routinely. CONCLUSIONS: These findings of low positive rates may be used to reassure veterinary staff that their exposure to cats does not appear to increase their risk of contracting toxoplasma infection and that pregnant women are not at an increased risk by owning a cat. PMID- 12769832 TI - Spectrum and antibiotic sensitivity of bacteria contaminating the upper gut in patients with malabsorption syndrome from the tropics. AB - BACKGROUND: Various causes of malabsorption syndrome (MAS) are associated with intestinal stasis that may cause small intestinal bacterial overgrowth (SIBO). Frequency, nature and antibiotic sensitivity of SIBO in patients with MAS are not well understood. METHODS: Jejunal aspirates of 50 consecutive patients with MAS were cultured for bacteria and colony counts and antibiotic sensitivity were performed. Twelve patients with irritable bowel syndrome were studied as controls. RESULTS: Culture revealed growth of bacteria in 34/50 (68%) patients with MAS and 3/12 controls (p < 0.05). Colony counts ranged from 3 x 10(2) to 10(15) (median 10(5)) in MAS and 100 to 1000 (median 700) CFU/ml in controls (p 0.003). 21/50 (42%) patients had counts GreaterEqual;105 CFU/ml in MAS and none of controls (p < 0.05). Aerobes were isolated in 34/34 and anaerobe in 1/34. Commonest Gram positive and negative bacteria were Streptococcus species and Escherichia coli respectively. The isolated bacteria were more often sensitive to quinolones than to tetracycline (ciprofloxacin: 39/47 and norfloxacin: 34/47 vs. tetracycline 19/47, <0.01), ampicillin, erythromycin and co-trimoxazole (21/44, 14/22 and 24/47 respectively vs. tetracycline, p = ns). CONCLUSIONS: SIBO is common in patients with MAS due to various causes and quinolones may be the preferred treatment. This needs to be proved further by a randomized controlled trial. PMID- 12769833 TI - Increased blood product use among coronary artery bypass patients prescribed preoperative aspirin and clopidogrel. AB - BACKGROUND: The administration of antiplatelet drugs before coronary artery bypass graft surgery (CABG) is associated with an increased risk of major hemorrhage and related surgical reexploration. Little is known about the relative effect of combined clopidogrel and aspirin on blood product use around the time of CABG. We evaluated the associated risk between the combined use of aspirin and clopidogrel and the transfusion of blood products perioperatively. METHODS: We retrospectively studied a cohort of 659 individuals who underwent a first CABG, without concomitant valvular or aortic surgery, at a single large Canadian cardiac surgical centre between January 2000 and April 2002. The four study exposure groups were those prescribed aspirin (n = 105), clopidogrel (n = 11), the combination of both (n = 46), or neither drug (n = 497), within 7 days prior to CABG. The primary study outcome was the excessive transfusion of blood products during CABG and up to the second post-operative day, defined as > or = 2 units of packed red blood cells (PRBC), > or = 2 units of fresh frozen plasma, > or = 5 units of cryoprecipitate or > or = 5 units of platelets. Secondary outcomes included the mean number of transfused units of each type of blood product. RESULTS: A greater mean number of units of PRBC were transfused among those who received clopidogrel alone (2.9) or in combination with aspirin (2.4), compared to those on aspirin alone (1.9) or neither antiplatelet drug (1.4) (P = 0.001). A similar trend was seen for the respective mean number of transfused units of platelets (3.6, 3.7, 1.3 and 1.0; P < 0.001) and fresh frozen plasma (2.5, 3.1, 2.3, 1.6; P = 0.01). Compared to non-users, the associated risk of excessive blood product transfusion was highest among recipients of aspirin and clopidogrel together (adjusted OR 2.2, 95% CI 1.1-4.3). No significant association was seen among lone users of aspirin (adjusted OR 1.0, 95% CI 0.6 1.6) or clopidogrel (adjusted OR 0.7, 95% CI 0.2-2.5), compared to non-users. CONCLUSIONS: While combined use of aspirin and clopidogrel shortly before CABG surgery may increase the associated risk of excess transfusion of blood products perioperatively, several study limitations prevent any confident conclusions from being drawn. Beyond challenging these findings, future research might focus on the value of both intraoperative monitoring of platelet function, and the effectiveness of antifibrinolytic agents, at reducing the risk of postoperative bleeding. PMID- 12769835 TI - A nuclear MAL-function links Rho to SRF. AB - The activation of SRF-mediated transcription in response to Rho GTPase-induced actin assembly involves the ability of SRF to sense changes in the cellular level of actin monomers. An article in the May 2 issue of Cell identifies the transcription factor MAL as an actin binding protein that functions as an SRF coactivator and whose translocation from cytoplasm to nucleus depends on its dissociation from actin monomers (Miralles et al., 2003). PMID- 12769834 TI - The kinase MSK1 is required for induction of c-fos by lysophosphatidic acid in mouse embryonic stem cells. AB - BACKGROUND: The regulation of the immediate-early gene c-fos serves as a paradigm for signal-activated gene induction. Lysophosphatidic acid is a potent serum borne mitogen able to induce c-fos. RESULTS: Analysing the signalling events following stimulation of mouse embryonic stem cells with serum and lysophosphatidic acid, we show that the extracellular signal-regulated kinase (ERK) pathway is involved in mediating c-fos induction. We demonstrate that the ERK-activated kinase MSK1 is required for full c-fos promoter activation, as well as for the phosphorylation of cAMP-responsive element (CRE) binding proteins. We propose that MSK1 contributes to ERK-mediated c-fos promoter activation by targeting CRE binding proteins. CONCLUSION: These results show that MSK1 is an important ERK-activated mediator of mitogen-stimulated c-fos induction. In addition, they indicate that MSK1 could act through CRE binding proteins to achieve c-fos promoter activation. Thus, they further our understanding of the complex regulation of the model immediate-early gene c-fos. PMID- 12769837 TI - The paradoxical relationship between NHEJ and telomeric fusion. AB - Recent results shed new light on the origin of fusion products observed in the destabilized chromosomes of cancer and related diseases. These findings define an unusual relationship between nonhomologous end joining (NHEJ) and telomere "capping," with identical proteins playing opposing roles. PMID- 12769836 TI - FEARless in meiosis. AB - Degradation of mitotic cyclins is critical for exit from mitosis. Recent studies in budding yeast address the role of cyclin degradation in meiosis. Cyclin stabilization in meiosis I interferes with anaphase I spindle disassembly but, surprisingly, does not halt progression into meiosis II. PMID- 12769838 TI - mRNA decay: x (XRN1) marks the spot. AB - Degradation of mRNA is a vital aspect of gene expression. In yeast, Dcp1p, Dcp2p, Lsm1-7p, and Xrn1p are required for mRNA decay and are localized within discrete cytoplasmic foci; in the May 2 issue of Science, Sheth and Parker provide compelling evidence that these foci represent sites for mRNA decay. PMID- 12769839 TI - Early formation of mRNP: license for export or quality control? AB - Eukaryotic mRNA is processed by enzymes and packaged with proteins within nuclei to generate functional messenger ribonucleoprotein (mRNP) particles. Processing and packaging factors can interact with mRNA cotranscriptionally to form an early mRNP. Erroneous mRNP formation leads to nuclear retention and degradation of the mRNA. It therefore appears that one function of cotranscriptional mRNP assembly is to discard aberrant mRNPs early in their biogenesis. Cotranscriptional mRNP assembly may also enable the transcription machinery to respond to improper mRNP formation. PMID- 12769841 TI - Alpha v beta 5 integrin-dependent programmed cell death triggered by a peptide mimic of annexin V. AB - The diverse cytoplasmic domain sequences within the various integrin subunits are critical for integrin-mediated signaling into the cell (outside-in signaling) and for activation of ligand binding affinity (inside-out signaling). Here we introduce an approach based on phage display technology to identify molecules that specifically interact with the cytoplasmic domain of the beta 5 integrin subunit. We show that a peptide selected for binding specifically to the beta 5 cytoplasmic domain (VVISYSMPD) induces apoptosis upon internalization. The cell death process induced by VVISYSMPD is sensitive to modulation by growth factors and by protein kinase C (PKC), and it cannot be triggered in beta 5 null cells. Finally, we show that the VVISYSMPD peptide is a mimic of annexin V. Our results suggest a functional link between the alpha v beta 5 integrin, annexin V, and programmed cell death. We propose the term "endothanatos" to designate this phenomenon. PMID- 12769840 TI - Type IV pilin structure and assembly: X-ray and EM analyses of Vibrio cholerae toxin-coregulated pilus and Pseudomonas aeruginosa PAK pilin. AB - Pilin assembly into type IV pili is required for virulence by bacterial pathogens that cause diseases such as cholera, pneumonia, gonorrhea, and meningitis. Crystal structures of soluble, N-terminally truncated pilin from Vibrio cholera toxin-coregulated pilus (TCP) and full-length PAK pilin from Pseudomonas aeruginosa reveal a novel TCP fold, yet a shared architecture for the type IV pilins. In each pilin subunit a conserved, extended, N-terminal alpha helix wrapped by beta strands anchors the structurally variable globular head. Inside the assembled pilus, characterized by cryo-electron microscopy and crystallography, the extended hydrophobic alpha helices make multisubunit contacts to provide mechanical strength and flexibility. Outside, distinct interactions of adaptable heads contribute surface variation for specificity of pilus function in antigenicity, motility, adhesion, and colony formation. PMID- 12769842 TI - Increased ezrin expression and activation by CDK5 coincident with acquisition of the senescent phenotype. AB - Passage of normal cells in culture leads to senescence, an irreversible cell cycle exit characterized by biochemical changes and a distinctive morphology. Cellular stresses, including oncogene activation, can also lead to senescence. Consistent with an anti-oncogenic role for this process, the tumor suppressor pRb plays a critical role in senescence. Reexpression of pRb in human tumor cells results in senescence-like changes including cell cycle exit and shape changes. Here we show that senescence is accompanied by increased expression and altered localization of ezrin, an actin binding protein involved in membrane-cytoskeletal signaling. pRb expression results in the stimulation of CDK5-mediated phosphorylation of ezrin with subsequent membrane association and induction of cell shape changes, linking pRb activity to cytoskeletal regulation in senescent cells. PMID- 12769843 TI - Skp2 regulates Myc protein stability and activity. AB - Myc is an oncoprotein transcription factor that plays a prominent role in cancer. Like many transcription factors, Myc is an unstable protein that is destroyed by ubiquitin (Ub)-mediated proteolysis. Here, we report that the oncoprotein and Ub ligase Skp2 regulates Myc ubiquitylation and stability. Because of the growing number of Ub ligases that function as transcriptional coactivators, we speculated that Skp2 might also regulate Myc's transcriptional activity. Consistent with this model, we also show that Skp2 is a transcriptional coactivator for Myc, recognizing an essential element within the Myc activation domain and activating Myc target genes. These data suggest that Skp2 functions to connect Myc activity and destruction, and reveal an unexpected oncoprotein connection that may play an important role in controlling cell growth in normal and cancer cells. PMID- 12769844 TI - The F-box protein Skp2 participates in c-Myc proteosomal degradation and acts as a cofactor for c-Myc-regulated transcription. AB - The transcription regulatory oncoprotein c-Myc controls genes involved in cell growth, apoptosis, and oncogenesis. c-Myc is turned over very quickly through the ubiquitin/proteasome pathway. The proteins involved in this process are still unknown. We have found that Skp2 interacts with c-Myc and participates in its ubiquitylation and degradation. The interaction between Skp2 and c-Myc occurs during the G1 to S phase transition of the cell cycle in normal lymphocytes. Surprisingly, Skp2 enhances c-Myc-induced S phase transition and activates c-Myc target genes in a Myc-dependent manner. Further, Myc-induced transcription was shown to be Skp2 dependent, suggesting interdependence between c-Myc and Skp2 in activation of transcription. Moreover, Myc-dependent association of Skp2, ubiquitylated proteins, and subunits of the proteasome to a c-Myc target promoter was demonstrated in vivo. The results suggest that Skp2 is a transcriptional cofactor for c-Myc and indicates a close relationship between transcription activation and transcription factor ubiquitination. PMID- 12769845 TI - Requirement of Skp1-Bub1 interaction for kinetochore-mediated activation of the spindle checkpoint. AB - The spindle checkpoint transiently prevents cell cycle progression of cells that have incurred errors or failed to complete steps during mitosis, including those involving kinetochore function. The molecular nature of the primary signal transmitted from defective kinetochores and how it is detected by the spindle checkpoint are unknown. We report biochemical evidence that Bub1, a component of the spindle checkpoint, associates with centromere (CEN) DNA via Skp1, a core kinetochore component in budding yeast. The Skp1's interaction with Bub1 is required for the mitotic delay induced by kinetochore tension defects, but not for the arrest induced by spindle depolymerization, kinetochore assembly defects, or Mps1 overexpression. We propose that the Skp1-Bub1 interaction is important for transmitting a signal to the spindle checkpoint pathway when insufficient tension is present at kinetochores. PMID- 12769846 TI - Contingent phosphorylation/dephosphorylation provides a mechanism of molecular memory in WASP. AB - Cells can retain information about previous stimuli to produce distinct future responses. The biochemical mechanisms by which this is achieved are not well understood. The Wiskott-Aldrich syndrome protein (WASP) is an effector of the Rho family GTPase Cdc42, whose activation leads to stimulation of the actin nucleating assembly, Arp2/3 complex. We demonstrate that efficient phosphorylation and dephosphorylation of WASP at Y291 are both contingent on binding to activated Cdc42. Y291 phosphorylation increases the basal activity of WASP toward Arp2/3 complex and enables WASP activation by new stimuli, SH2 domains of Src-family kinases. The requirement for contingency in both phosphorylation and dephosphorylation enables long-term storage of information by WASP following decay of GTPase signals. This biochemical circuitry allows WASP to respond to the levels and timing of GTPase and kinase signals. It provides mechanisms to specifically achieve transient or persistent actin remodeling, as well as long-lasting potentiation of actin-based responses to kinases. PMID- 12769847 TI - Phosphorylation of the WASP-VCA domain increases its affinity for the Arp2/3 complex and enhances actin polymerization by WASP. AB - Wiskott-Aldrich syndrome protein (WASP) and neural (N)-WASP regulate dynamic actin structures through the ability of their VCA domains to bind to and stimulate the actin nucleating activity of the Arp2/3 complex. Here we identify two phosphorylation sites in the VCA domain of WASP at serines 483 and 484. S483 and S484 are substrates for casein kinase 2 in vitro and in vivo. Phosphorylation of these residues increases the affinity of the VCA domain for the Arp2/3 complex 7-fold and is required for efficient in vitro actin polymerization by the full length WASP molecule. We propose that constitutive VCA domain phosphorylation is required for optimal stimulation of the Arp2/3 complex by WASP. PMID- 12769848 TI - Crystal structure of a tetradecameric assembly of the association domain of Ca2+/calmodulin-dependent kinase II. AB - We report the crystal structure of the 143 residue association domain of Ca(2+)/calmodulin-dependent protein kinase II (CaMKII). The association domain forms a hub-like assembly, composed of two rings of seven protomers each, which are stacked head to head and held together by extensive interfaces. The tetradecameric organization of the assembly was confirmed by analytical ultracentrifugation and multiangle light scattering. Individual protomers form wedge-shaped structures from which N-terminal helical segments that connect to the kinase domain extend toward the equatorial plane of the assembly, consistent with the arrangement of the kinase domains in a second outer ring. A deep and highly conserved pocket present within the association domain may serve as a docking site for proteins that interact with CaMKII. PMID- 12769849 TI - Computational and experimental identification of C. elegans microRNAs. AB - MicroRNAs (miRNAs) constitute an extensive class of noncoding RNAs that are thought to regulate the expression of target genes via complementary base-pair interactions. To date, cloning has identified over 200 miRNAs from diverse eukaryotic organisms. Despite their success, such biochemical approaches are skewed toward identifying abundant miRNAs, unlike genome-wide, sequence-based computational predictions. We developed informatic methods to predict miRNAs in the C. elegans genome using sequence conservation and structural similarity to known miRNAs and generated 214 candidates. We confirmed the expression of four new miRNAs by Northern blotting and used a more sensitive PCR approach to verify the expression of ten additional candidates. Based on hypotheses underlying our computational methods, we estimate that the C. elegans genome may encode between 140 and 300 miRNAs and potentially many more. PMID- 12769850 TI - MOF-regulated acetylation of MSL-3 in the Drosophila dosage compensation complex. AB - Dosage compensation ensures equal expression of X-linked genes in males and females. In Drosophila, equalization is achieved by hypertranscription of the male X chromosome. This process requires an RNA/protein containing dosage compensation complex (DCC). Here we use RNA interference of individual DCC components to define the order of complex assembly in Schneider cells. We show that interaction of MOF with MSL-3 leads to specific acetylation of MSL-3 at a single lysine residue adjacent to one of its chromodomains. We observe that localization of MSL-3 to the X chromosome is RNA dependent and acetylation sensitive. We find that the acetylation status of MSL-3 determines its interaction with roX2 RNA. Furthermore, we find that RPD3 interacts with MSL-3 and that MSL-3 can be deacetylated by the RPD3 complex. We propose that regulated acetylation of MSL-3 may provide a mechanistic explanation for spreading of the dosage compensation complex along the male X chromosome. PMID- 12769851 TI - RNAse III-mediated degradation of unspliced pre-mRNAs and lariat introns. AB - Double-stranded RNA (dsRNA) has emerged as a modulator of gene expression, from gene silencing to antiviral responses. Here we show that dsRNA stem-loop structures found in intronic regions of the S. cerevisiae RPS22B and RPL18A transcripts trigger degradation of unspliced pre-mRNAs and lariat introns and can control the level of mRNA produced from these intron-containing genes. The dsRNA regions are cleaved by Rnt1p, the yeast ortholog of RNase III, which creates an entry site for complete degradation by the Xrn1p and Rat1p exonucleases and by the nuclear exosome. These results identify an alternative discard pathway for precursors and products of the splicing machinery and a physiological function for dsRNA in eukaryotic RNA catabolism. PMID- 12769852 TI - Transcription of Leishmania major Friedlin chromosome 1 initiates in both directions within a single region. AB - Almost nothing is known about the sequences involved in transcription initiation of protein-coding genes in the parasite Leishmania. We describe here the transcriptional analysis of chromosome 1 (chr1) from Leishmania major Friedlin (LmjF) which encodes the first 29 genes on one DNA strand, and the remaining 50 on the opposite strand. Strand-specific nuclear run-on assays showed that a low level of nonspecific transcription probably takes place over the entire chromosome, but an approximately 10-fold higher level of coding strand-specific RNA polymerase II (Pol II)-mediated transcription initiates within the strand switch region. 5' RACE studies localized the initiation sites to a <100 bp region. Transfection studies support the presence of a bidirectional promoter within the strand-switch region, but suggest that other factors are also involved in Pol II transcription. Thus, while in most eukaryotes each gene possesses its own promoter, a single region seems to drive the expression of the entire chr1 in LmjF. PMID- 12769853 TI - Independent recruitment in vivo by Gal4 of two complexes required for transcription. AB - We use a modified form of ChIP to analyze the recruitment of seven sets of proteins to the yeast GAL genes upon induction. We resolve three stages of recruitment: first SAGA, then Mediator, and finally Pol II along with four other proteins (including TBP) bind the promoter. In a strain lacking SAGA, Mediator is recruited with a time course indistinguishable from that observed in wild-type cells. Our results are consistent with the notion that a single species of activator, Gal4, separately contacts, and thereby directly recruits, SAGA and Mediator. PMID- 12769854 TI - Distinct strategies to make nucleosomal DNA accessible. AB - One hallmark of ATP-dependent remodeling complexes is the ability to make nucleosomal DNA accessible to regulatory factors. We have compared two prominent human ATP-dependent remodelers, BRG1 from the SWI/SNF family and SNF2h from the ISWI family, for their abilities to make a spectrum of nucleosomal sites accessible. By measuring rates of remodeling at seven different sites on a mononucleosome and at six different sites on the central nucleosome of a trinucleosome, we have found that BRG1 opens centrally located sites more than an order of magnitude better than SNF2h. We provide evidence that this capability of BRG1 is caused by its ability to create DNA loops on the surface of a nucleosome, even when that nucleosome is constrained by adjacent nucleosomes. This specialized ability to make central sites accessible should allow SWI/SNF family complexes to facilitate binding of nuclear factors in chromatin environments where adjacent nucleosomes might otherwise constrain mobility. PMID- 12769855 TI - A central role for DNA replication forks in checkpoint activation and response. AB - The checkpoint proteins Rad53 and Mec1-Ddc2 regulate many aspects of cell metabolism in response to DNA damage. We have examined the relative importance of downstream checkpoint effectors on cell viability. Checkpoint regulation of mitosis, gene expression, and late origin firing make only modest contributions to viability. By contrast, the checkpoint is essential for preventing irreversible breakdown of stalled replication forks. Moreover, recruitment of Ddc2 to nuclear foci and subsequent activation of the Rad53 kinase only occur during S phase and require the assembly of replication forks. Thus, DNA replication forks are both activators and primary effectors of the checkpoint pathway in S phase. PMID- 12769856 TI - RecFOR proteins load RecA protein onto gapped DNA to accelerate DNA strand exchange: a universal step of recombinational repair. AB - Genetic evidence suggests that the RecF, RecO, and RecR (RecFOR) proteins participate in a common step of DNA recombination and repair, yet the biochemical event requiring collaboration of all three proteins is unknown. Here, we show that the concerted action of the RecFOR complex directs the loading of RecA protein specifically onto gapped DNA that is coated with single-stranded DNA binding (SSB) protein, thereby accelerating DNA strand exchange. The RecFOR complex recognizes the junction between the ssDNA and dsDNA regions and requires a base-paired 5' terminus at the junction. Thus, the RecFOR complex is a structure-specific mediator that targets recombinational repair to ssDNA-dsDNA junctions. This reaction reconstitutes the initial steps of recombinational gapped DNA repair and uncovers an event also common to the repair of ssDNA-tailed intermediates of dsDNA-break repair. We propose that the behavior of the RecFOR proteins is mimicked by functional counterparts that exist in all organisms. PMID- 12769857 TI - Modular architecture of the bacteriophage T7 primase couples RNA primer synthesis to DNA synthesis. AB - DNA primases are template-dependent RNA polymerases that synthesize oligoribonucleotide primers that can be extended by DNA polymerase. The bacterial primases consist of zinc binding and RNA polymerase domains that polymerize ribonucleotides at templating sequences of single-stranded DNA. We report a crystal structure of bacteriophage T7 primase that reveals its two domains and the presence of two Mg(2+) ions bound to the active site. NMR and biochemical data show that the two domains remain separated until the primase binds to DNA and nucleotide. The zinc binding domain alone can stimulate primer extension by T7 DNA polymerase. These findings suggest that the zinc binding domain couples primer synthesis with primer utilization by securing the DNA template in the primase active site and then delivering the primed DNA template to DNA polymerase. The modular architecture of the primase and a similar mechanism of priming DNA synthesis are likely to apply broadly to prokaryotic primases. PMID- 12769858 TI - Distinct biogenesis pathways for human telomerase RNA and H/ACA small nucleolar RNAs. AB - Functional RNAs are processed according to cues from precursor expression context. The presence of an H/ACA motif within the human telomerase RNA (hTR) suggested that telomerase might utilize the biogenesis pathway of an H/ACA small nucleolar RNA. Here, we have investigated the requirements for processing, stability, and function of hTR. Instead of a biogenesis pathway dependent on transcription context or precursor sequence, we find that maturation of hTR requires cooperation of the H/ACA motif and an additional motif unique to the hTR H/ACA domain. This telomerase-specific hTR element is mutated in dyskeratosis congenita, and the disease-associated hTR substitution impairs RNA accumulation. Disease-associated hTR variants with sequence changes outside the H/ACA domain do not affect RNA processing or stability; they instead impose a catalytic defect. Our results reveal differences between the biogenesis of hTR and other H/ACA motif RNAs and demonstrate distinct mechanisms of telomerase inhibition in human disease. PMID- 12769859 TI - NEJ1 prevents NHEJ-dependent telomere fusions in yeast without telomerase. AB - In a search for genes involved in cell-type-dependent chromosome instability, we have found a role for NEJ1, a regulator of nonhomologous end joining (NHEJ), in cells that survive in the absence of telomerase. In yeast, NHEJ is regulated by mating-type status through NEJ1, which is repressed in a/alpha cells. For efficient NHEJ, NEJ1 is required as part of a complex with LIF1 and DNL4, which catalyzes DNA ligation. In haploid cells without telomerase, we find that the absence of NEJ1 results in high frequencies of circular chromosomes in type II survivors (i.e., those typified by lengthened telomere repeat tracts). These telomere fusion events are DNL4 dependent. NEJ1 therefore has a role in protecting telomeres from end fusions by NHEJ in the absence of telomerase that contrasts with its role in promoting repair at sites of DNA double-strand breaks. PMID- 12769860 TI - Telomerase and ATM/Tel1p protect telomeres from nonhomologous end joining. AB - Telomeres protect chromosome ends from fusing to double-stranded breaks (DSBs). Using a quantitative real-time PCR assay, we show that nonhomologous end joining between a telomere and an inducible DSB was undetectable in wild-type cells, but occurred within a few hours of DSB induction in approximately 1/2000 genomes in telomerase-deficient cells and in >1/1000 genomes in telomerase-deficient cells also lacking the ATM homolog Tel1p. The fused telomeres contained very little telomeric DNA, suggesting that catastrophic telomere shortening preceded fusion. Lengthening of telomeres did not prevent such catastrophic telomere shortening and fusion events. Telomere-DSB fusion also occurred in cells containing a catalytically inactive telomerase and in tel1 mec1 cells where telomerase cannot elongate telomeres. Thus, telomerase and Tel1p function in telomere protection as well as in telomere elongation. PMID- 12769861 TI - Opposed regulation of corepressor CtBP by SUMOylation and PDZ binding. AB - The transcription corepressor CtBP is often recruited to the target promoter via interaction with a conserved PxDLS motif in the interacting repressor. In this study, we demonstrate that CtBP1 was SUMOylated and that its SUMOylation profoundly affected its subcellular localization. SUMOylation occurred at a single Lys residue, Lys428, of CtBP1. CtBP2, a close homolog of CtBP1, lacked the SUMOylation site and was not modified by SUMO-1. Mutation of Lys428 into Arg (K428R) shifted CtBP1 from the nucleus to the cytoplasm, while it had little effect on its interaction with the PxDLS motif. Consistent with a change in localization, the K428R mutation abolished the ability of CtBP1 to repress the E cadherin promoter activity. Notably, SUMOylation of CtBP1 was inhibited by the PDZ domain of nNOS, correlating with the known inhibitory effect of nNOS on the nuclear accumulation of CtBP1. This study identifies SUMOylation as a regulatory mechanism underlying CtBP1-dependent transcriptional repression. PMID- 12769862 TI - Drosophila sex-lethal inhibits the stable association of the 40S ribosomal subunit with msl-2 mRNA. AB - The inhibition of male-specific lethal-2 (msl-2) mRNA translation in female flies is essential for X chromosome dosage compensation in Drosophila melanogaster. Translational repression of msl-2 requires sex-lethal (SXL) binding to uridine rich sequences in both the 5' and 3' untranslated regions (UTRs) of the message. We delineate the msl-2 mRNA sequence elements that are important for regulation by SXL and identify functionally critical sequences adjacent to regulatory SXL binding sites. We demonstrate that SXL inhibits translation initiation and prevents the stable association of the 40S ribosomal subunit with the mRNA in a manner that does not require the presence of a cap structure at the 5' end of the mRNA. These results elucidate a critical regulatory step for dosage compensation in Drosophila melanogaster. PMID- 12769863 TI - An NMD pathway in yeast involving accelerated deadenylation and exosome-mediated 3'-->5' degradation. AB - Eukaryotic mRNAs containing premature termination codons are subjected to accelerated turnover, known as nonsense-mediated decay (NMD). Recognition of translation termination events as premature requires a surveillance complex, which includes the RNA helicase Upf1p. In Saccharomyces cerevisiae, NMD provokes rapid decapping followed by 5'-->3' exonucleolytic decay. Here we report an alternative, decapping-independent NMD pathway involving deadenylation and subsequent 3'-->5' exonucleolytic decay. Accelerated turnover via this pathway required Upf1p and was blocked by the translation inhibitor cycloheximide. Degradation of the deadenylated mRNA required the Rrp4p and Ski7p components of the cytoplasmic exosome complex, as well as the putative RNA helicase Ski2p. We conclude that recognition of NMD substrates by the Upf surveillance complex can target mRNAs to rapid deadenylation and exosome-mediated degradation. PMID- 12769866 TI - A possible mechanism for the physiological suppression of conspecific eggs and larvae following superparasitism by solitary endoparasitoids. AB - Competition for possession of a host by internal solitary parasitoids has been attributed to physical combat and physiological suppression, but the mechanisms that result in what has been referred to as physiological suppression is poorly understood. Some insights are provided by the studies reported here using the solitary endoparasitoid, Campoletis sonorensis (Cameron). Embryos of C. sonorensis less than ten hours old rarely hatch in various artificial media, while embryos twenty hours or older generally hatch. These results suggest that young embryos in which the embryonic membranes have not yet formed are only able to develop in a narrow range of environments represented by the nonparasited hemolymph. In contrast, embryos in which the embryonic membranes are formed are able to develop in a wide range of environments represented by parasitized hemolymph which has been shown by a number of studies to change. These ideas were given support by studies reported here, where young and older eggs were incubated singly or paired. We suggest the general changes in the hemolymph of a parasitized host become unfavorable for the development of newly oviposited eggs. PMID- 12769867 TI - Interaction of calyx fluid and venom from Microplitis croceipes (Braconidae) on developmental disruption of the natural host, Heliocoverpa zea, and two atypical hosts, Galleria mellonella and Spodoptera exigua. AB - Polydnaviruses of many braconid and ichneumonid endoparasitoids play an important role in the successful parasitism of their hosts. The host's development is altered and its immune response is also suppressed. In this study, we compared the effects of calyx fluid and venom on the development of the natural host, Helicoverpa zea, and two atypical hosts that the parasitoid does not normally attack in nature, Galleria mellonella and Spodoptera exigua. The levels of calyx fluid and?or venom injected was 0.05, 0.1 and 0.2 female equivalents (FE)/larva. In H. zea, calyx fluid significantly reduced larval growth on day 5 post injection. Venom alone did not affect larval growth but it synergized the action of calyx fluid by reducing growth earlier and for a longer period after injection. Other effects of calyx fluid on the host, either alone or in combination with venom, were an increase in developmental period, and a reduction in percent emergence and weight of adult moths. The percentage of H. zea larvae that pupated was not affected by calyx fluid or venom. In Galleria mellonella, venom alone reduced larval growth comparable to calyx fluid and both tissues induced the effects on day 1 post injection. Other effects caused by calyx fluid or venom alone or the combination were a reduction in percent pupation and emergence, and the average adult weight. In S. exigua, high mortality occurred when 4th instar larvae were injected. Although the injection of larger fifth instars reduced overall mortality, the sham-injected larvae only gained weight during the first 24 hours after injection (from day 0 to day 1). However, adults were produced at all doses of calyx fluid or venom. The effects of the virus on development in this species were a prolongation of the larval stage and reduction of adult weight by calyx fluid in combination with venom. In conclusion, injections of calyx fluid and venom of Microplitis croceipes can differentially affect the growth and development of its natural host H. zea, and atypical host, G. mellonella, but only a minimal effect was observed in S. exigua. PMID- 12769868 TI - Host/parasitoid interactions: critical timing of parasitoid-derived products. AB - Short-term in vitro incubations were used to examine the ability of endoparasitoid larvae to produce and release both ecdysteroids and proteins into their environment. Second instar larvae of both Chelonus near curvimaculatus and Ascogaster quadridentata were observed by SDS-PAGE to release temporally-similar polypeptides in the 20-30kD M(r) range. Peak occurrence of these polypeptides coincided with shedding of the anal vesicle, immediately prior to ecdysis to the third instar. Ecdysis also coincided with the switch from endoparasitic to ectoparasitic development in vivo. Polyclonal antibodies were generated against a particular 27kD polypeptide of Chelonus, which was found to be species-specific and localized primarily within the anal vesicle during the latter part of the second stadium and whole body homogenates of third instars. In vitro incorporation studies using (35)S-methionine indicated rapid changes in the synthetic abilities of second instar larvae shortly before ecdysis. The production and release of ecdysteroids, as measured by RIA, was found to precede the peak occurrence of the 27kD polypeptide and ecdysteroid presence was undetectable following the molt. In contrast, the polypeptides were observed to gradually increase prior to the molt and slowly decrease after the molt. The Chelonus polypeptide was not detected in host tissues until after parasitoid egression. PMID- 12769869 TI - Parasitic castration of Pseudaletia separata by Cotesia kariyai and its association with polydnavirus gene expression. AB - Parasitization by the endoparasitoid Cotesia kariyai caused the inhibition of spermatogenesis of Pseudaletia separata. This phenomenon is called parasitic castration. The degree of castration was dependent on the host stage parasitized. Host parasitized on day 1 of the 4th stadium (the time of primary spermatocyte accumulation), had testicular cells with abnormal chromosomes appearing two days after parasitization, and spermiogenesis was completely inhibited. However, when hosts were parasitized on day 0 of the 6th (final) stadium, the degree of castration was less severe, and elongated cells appeared similar to those found in nonparasitized larvae. Results of this study involving injection of C. kariyai polydnavirus (CkPV) and venom suggested that these wasp components caused the appearance of abnormal chromosomes in specific germ cells, which were in mitotic or meiotic prophases. The amount of CkPV gene expression in host testes increased immediately after parasitization and reached a maximum 12h later. The early expressed CkPV gene(s) may be related to the parasitic castration phenomenon. PMID- 12769870 TI - Aberrant nutritional regulation of carbohydrate synthesis by parasitized Manduca sexta L. AB - The present studies confirm that storage carbohydrate synthesis from [1 (13)C]glucose is elevated in Manduca sexta parasitized by Cotesia congregata, despite a decrease in the rate of metabolism of the labeled substrate. Further, the results demonstrate that a similar pattern of carbohydrate synthesis and glucose metabolism was induced in normal larvae by administration of the glycolytic inhibitor, iodoacetate. (13)C enrichment of C6 of trehalose and glycogen demonstrated randomization of the C1 label at the triose phosphate step of the glycolytic/gluconeogenic pathway and suggested that gluconeogenesis, that is, de novo carbohydrate formation, contributed to the synthesis of carbohydrate in both normal and parasitized insects. Accounting for differences in the (13)C enrichment in C1 of trehalose and glycogen due to direct labeling from [1 (13)C]glucose, the mean C6/C1 labeling ratios in trehalose and glycogen of parasitized larvae and insects treated with iodoacetate were greater than the mean ratio observed in normal larvae, suggesting a greater contribution of gluconeogenesis to trehalose labeling in parasitized insects. This conclusion was confirmed by additional investigations on the metabolism of [3-(13)C]alanine by normal and parasitized insects. The pattern of (13)C enrichment in hemolymph trehalose observed in normal larvae maintained on a low carbohydrate diet indicated a large contribution of gluconeogenesis, while gluconeogenesis contributed very little to trehalose labeling in normal insects maintained on a high carbohydrate diet. Parasitized insects maintained on a high or a low carbohydrate diet displayed a significantly greater contribution of gluconeogenesis to trehalose labeling than was observed in normal larvae maintained on the same diets. In conclusion, these investigations indicate that regulation over the utilization of dietary glucose for trehalose and glycogen synthesis as well as the dietary regulation of de novo carbohydrate synthesis were altered by parasitism. PMID- 12769871 TI - Preultimate 4th/5th instar Trichoplusia ni naturally-injected with venom/calyx fluid from Chelonus curvimaculatus precociously metamorphose, rather than obey the metamorphic size threshold that would normally compel molting to a 5th/6th instar. AB - In normally regulated larval metamorphosis of Trichoplusia ni, a 4th, 5th or other numbered instar is a 'preultimate' instar, and will normally continue larval molting, if the larva has not yet surpassed the critical (minimal) size threshold corresponding to attainment of the 'ultimate' (metamorphic) instar. Natural injection of T. ni embryos with venom/calyx fluid of female Chelonus sp. near curvimaculatus caused 'penultimate' 4th or 5th instar larvae that would normally molt at least once more, to a 5th/6th instar, to instead precociously metamorphose without another larval molt. These effects were observed in naturally-injected insects that never contained either a parasite larva, a viable parasite embryo, or a parasite egg. These data demonstrate that this effect of venom/calyx fluid of this wasp to induce precocious metamorphosis, at an instar earlier than would otherwise have typically occurred under normal growth conditions, does not require the presence of a parasite larva. Other data did indicate the parasite larva contributes an additional effect that causes a 4th instar host (that from its size would normally require not just one, but at least two more larval molts to reach the metamorphic instar) to not grow to the size metamorphic threshold, but to instead, precociously metamorphose at an even smaller size than occurs with the venom/calyx fluid alone. Additionally, arylphorin was precociously highly expressed in parasitized hosts in a manner independent of a decline in the host JH titer. Therefore, the main target of the venom/calyx fluid activity to induce precocious metamorphosis appears to be an event upstream of the decline in JH production by the corpora allata. Pseudoparasitized hosts become developmentally arrested as precocious prepupae and express a 2.7kb polydnavirus transcript. The larger (but still subthreshold size) larvae showed less suppressed prepupal ecdysteroid titers, less developmental suppression, and a much weaker expression of that transcript. A general model for mechanisms of action of chelonine venom/calyx fluid, and larvae, to cause precocious host metamorphosis and suppressed prepupal development is presented that is based on the current 'size threshold' model of normal lepidopteran development, rather than the older, displaced 'instar count' model. By basing the model for chelonine regulation of host development on the current 'size threshold' model for normal development, the proposed model for chelonine action both accounts for observations reported on various species of that subfamily and makes useful, testable predictions. PMID- 12769872 TI - Microplitis croceipes teratocytes: in vitro culture and biological activity of teratocyte secreted protein. AB - Teratocytes originate from the dissociation of the extraembryonic serosal membrane in some Braconidae and Scelionidae. Methods used to culture teratocytes in vitro are described and the yield of teratocyte secreted proteins (TSP) was measured. Although 90% are viable after 6 days, in vitro teratocytes reached only half the diameter (32&mgr;m) of the same age teratocytes obtained in vivo. Teratocytes cultured in vitro secrete as much as 0.7&mgr;g of protein per day per larval equivalent ( approximately 900 cells). Presence of parasitoid larvae enhanced teratocyte viability while periodic exchange of medium did not. However, medium exchange significantly increased the total amount of protein secreted. Size and viability were improved with the addition of 10% FBS to the Ex-cell 400 culture medium. Non-denaturing PAGE showed at least 15 proteins with molecular sizes estimated to be between 24 to 347kDa in medium containing teratocytes. An in vitro fat body assay was developed to measure the effect of TSP on protein synthesis and juvenile hormone esterase (JHE) activity. Crude TSP inhibited in vitro incorporation of [(35)S]-methionine into protein synthesized by the fat body. The amount of JHE released from in vitro fat body treated with crude TSP was significantly less than controls, most likely caused by the inhibition of general protein synthesis. The active fraction of TSP passed through a 30kDa molecular weight cutoff filter but was retained by a 3kDa filter. SDS-PAGE revealed four proteins with molecular weights between 8 and 20kDa not present in control medium incubated without teratocytes. PMID- 12769873 TI - Host regulation effects of ovary fluid and venom of Aphidius ervi (Hymenoptera: Braconidae). AB - The host regulation effects of venom and ovary fluid of the endophagous braconid Aphidius ervi Haliday on the pea aphid, Acyrthosiphon pisum (Harris), have been studied. Extracts of ovaries and of venom glands were injected into nonparasitized 4th instar pea aphids, both separately and mixed. Aphids treated with parasitoid material died as 4th instars, often showing developmental arrest. In contrast, control aphids that received an injection of Pringle's saline solution regularly moulted to the adult stage and reproduced. Venom alone was as effective as the combined extracts in determining developmental arrest and death. Separate heat and protease treatments of these parasitoid's reproductive secretions significantly reduced their biological activity, suggesting that the active component(s) involved is a protein(s). SDS-PAGE analysis of haemolymph samples obtained from pea aphids which had received an injection of combined venom and ovary extract revealed an increase of the titre of various proteins, particularly in the 43-47kDa interval, as registered for truly parasitized hosts. This altered protein profile was first detected 48h following injection. Based on this information a tentative physiological model is proposed. The apical tract of host ovarioles, where the germarium and growing oocytes are located, is suggested to be one of the major targets of female parasitoid secretions injected with the egg. PMID- 12769874 TI - Relationships between polydnavirus genomes and viral gene expression. AB - Polydnavirus genomes and viral gene functions are atypical for viruses. Polydnaviruses are the only group of viruses with segmented DNA genomes and have an unusual obligate mutualistic association with parasitic Hymenoptera, in which the virus is required for survival of the wasp host and vice versa. The virus replicates asymptomatically in the wasp host but severely disrupts lepidopteran host physiology in the absence of viral DNA replication. It is not surprising then that viral gene expression is divergent in its two insect hosts and that differences in viral gene expression are linked to these divergent functions. Some viral genes are expressed only in the wasp host while other viral genes are expressed only in the lepidopteran host and are presumed to be involved in the disruption of host physiological systems. Our laboratory has described the expression and regulation of a family of viral genes implicated in suppressing the lepidopteran immune system, the cys-motif genes. In conjunction with these studies we have described the physical organization of additional viral gene segments. We have cloned, mapped and begun the sequence analysis of selected viral DNA segments. We have noted that some viral DNA segments are nested and that nested viral DNA segments encode the abundantly expressed, secreted cys motif genes. Conversely, other viral segments are not nested, encode less abundantly expressed genes and may be targeted intra-cellularly. These results suggest that nesting of segments in polydnavirus genomes may be linked to the levels of gene expression. By extension, the unique, segmented organization of polydnavirus genomes may be associated, in part, with the requirement for divergent levels of viral gene expression in lepidopteran hosts in the absence of viral DNA replication. PMID- 12769875 TI - A limited role in parasitism for Microplitis demolitor polydnavirus. AB - Spodoptera frugiperda larvae stung by Microplitis demolitor undergo physiological alterations characteristic of parasitism. However, despite these physiological modifications, parasitized S. frugiperda larvae never yield adult wasps. Our original hypothesis that unsuccessful parasitism was due to a transcriptionally inactive polydnavirus proved untrue. Microplitis demolitor polydnavirus (MdPDV) successfully infected and expressed, albeit transiently, in S. frugiperda hemocytes. MdPDV expression was most abundant in the first three days of parasitism, then sharply declined on Day 4 post-parasitization and continued to decline for the remainder of the study. During the period of MdPDV expression, S. frugiperda hemocytes were non-adherent, incapable of spreading in vitro and did not encapsulate M. demolitor eggs in vivo. Concurrent with diminishing viral expresssion, S. frugiperda hemocytes regained their ability to adhere and spread in vitro and encapsulated M. demolitor eggs in vivo. Although MdPDV disrupted S. frugiperda's encapsulation response for the first three days post-parasitization, M. demolitor was unable to develop in this noctuid species. Failure to develop was independant of viral activity, all M. demolitor eggs oviposited in S. frugiperda larvae failed to complete embryogenesis and died within 24 hour of oviposition. S. frugiperda larvae infected with MdPDV exhibited alterations in development very similar to other lepidopterans that are permissive hosts for M. demolitor. In addition, MdPDV DNA persisted in Spodoptera frugiperda hemocytes in the absence of viral expression. PMID- 12769876 TI - Hemocyte load and immune resistance to Asobara tabida are correlated in species of the Drosophila melanogaster subgroup. AB - Larvae from six Drosophila species of the melanogaster subgroup were compared for both the hemolymph concentration of hemocytes and the ability to encapsulate the eggs of the parasitoid Asobara tabida (Hymenoptera; Braconidae). Results showed a high correlation between the parasitized hosts' concentration of circulating hemocytes and their aptitude to form a hemocytic capsule around the parasitic eggs. Two conditions seem to be required for the encapsulation of A. tabida eggs to succeed: one condition, which may relate to the recognition of the parasite by the host defense system, is the occurrence of a primary hemocytic response, which gives rise to the amplification of the hemocyte population; the other condition is the presence in the parasitized hosts of a hemocyte load large enough for the cellular capsule to be completed before the parasitic egg becomes protected by embedment within the host tissues. Since the concentration in hemocytes of the parasitized hosts is partially related to the concentration in hemocytes before parasitization, Drosophila species carrying a high hemocyte load could be better predisposed to resist A. tabida. Results are discussed in regard to the importance of a non-specific, quantitative character, such as the host hemocyte load, for the co-evolutionary immune interactions between A. tabida and its Drosophila hosts. PMID- 12769877 TI - Overview of parasitism associated effects on host haemocytes in larval parasitoids and comparison with effects of the egg-larval parasitoid Chelonus inanitus on its host Spodoptera littoralis. AB - In the first part we review the effects of larval endoparasitoids and their polydnavirus and venom on the immune system of their hosts. In all systems investigated, haemocyte spreading and encapsulation activity was reduced; in some cases effects on total (THC) or differential (DHC) haemocyte count as well as modification of haemocyte morphology and ultrastructure were also documented. In many cases polydnavirus (and venom) were shown to play a major role in abrogation of the host's immune reaction. In the second part we present the first investigation of effects of parasitism and polydnavirus/venom on the immune system of the host for an egg-larval parasitoid, Chelonus inanitus. We observed that in 4th and 5th instar larvae, i.e. 7 to 10 days after parasitization, neither haemocyte spreading and encapsulation activity, nor DHC, nor haemocyte ultrastructure were altered. After parasitization with X-ray irradiated wasps, which inject polydnavirus and venom and infertile eggs, there was no alteration of the above mentioned parameters. Nevertheless, parasitoid larvae implanted into 4th instar larvae which developed from eggs parasitized with X-ray irradiated wasps were not encapsulated, whereas co-injected latex beads were. These results show that parasitism by this egg-larval parasitoid does not generally suppress the host's immune system but that polydnavirus/venom injected at oviposition prevent, by, as yet unknown mechanisms, encapsulation of the parasitoid larva. PMID- 12769878 TI - Manipulation of fifth-instar host (Manduca sexta) ecdysteroid levels by the parasitoid wasp Cotesia congregata. AB - Although 5th (last) instar parasitized Manduca sexta larvae undergo developmental arrest and do not wander, they exhibit a small hemolymph ecdysteroid peak (300 400pg/&mgr;l) which begins one day prior to the parasitoid's molt to the 3rd (last) instar and concomitant emergence from the host. Ecdysteroids present in this peak were 20-hydroxyecdysone, 20,26-dihydroxyecdysone and one or more very polar ecdysteroids, as well as small amounts of 26-hydroxyecdysone and ecdysone. In parasitized day-1 and -2 5th instars ligated just behind the 1st abdominal proleg, hemolymph ecdysteroid levels increased in both anterior and posterior portions (100-500pg/&mgr;l), while in unparasitized larvae, hormone levels only increased in the anterior portion (100-350pg/&mgr;l). Thus, the ecdysteroid peak observed in host 5th instars was probably produced, at least in part, by the parasitoids. It may serve to promote Cotesia congregata's molt from the second to the third instar and/or to facilitate parasitoid emergence from the host. In parasitized day-1 and -2 5th instars ligated between the last thoracic and 1st abdominal segments, hemolymph ecdysteroid titers reached much higher levels (500 3500pg/&mgr;l) in the anterior portion (no parasitoids present) than in the posterior portion (150-450pg/&mgr;l). Therefore, it appears that the parasitoid's regulation of hemolymph ecdysteroid titers occurs at two levels. First, parasitization neutralizes the host's ability to maintain its normal hemolymph ecdysteroid levels. Second, in a separate action, the parasitoid manipulates the ecdysteroid-producing machinery so that hemolymph levels are maintained at the 200-400pg/&mgr;l characteristic of day 3-4 hosts. This is the first report of a parasitoid's ability to interfere with the normal inhibitory mechanisms which prevent prothoracic gland production of ecdysteroid at inappropriate periods of insect growth and development. PMID- 12769879 TI - Prothoracic gland inactivation in Heliothis virescens (F.) (Lepidoptera:Noctuidae) larvae parasitized by Cardiochiles nigriceps Viereck (Hymenoptera:Braconidae). AB - Heliothis virescens (F.) last instar larvae parasitized by the endophagous braconid Cardiochiles nigriceps Viereck fail to attain the pupal stage, due to a parasitoid-induced alteration of ecdysteroid biosynthesis and metabolism. Currently available information on host prothoracic gland inactivation in this host-parasitoid system is reported here. Prothoracic glands of H. virescens mature larvae show a depressed biosynthetic activity, without undergoing gross morphological disruption. The ultrastructure of gland cells is characterized by minor parasitoid-induced changes, with the rough endoplasmic reticulum appearing more developed and electrondense than in nonparasitized controls. Eventually, the cells of prothoracic glands of parasitized host last instar larvae die but maintain their structural integrity. The inactivation of pupally committed host prothoracic glands is achieved through the disruption of the PTTH signal transduction pathway. The second messenger cAMP appears to be normally produced in response to PTTH stimulation of glands explanted from parasitized host larvae, however the downstream activation of the cAMP-dependent protein kinase does not appear to occur. In fact, a marked underphosphorylation of regulatory target proteins is observed. This underphosphorylation is associated with a significant reduction in general protein synthesis, which appears to be blocked at the translational level, to a redirection of specific protein synthesis and to a drastic suppression of ecdysteroidogenesis. These parameters appeared to be correlated in a kinetic time-course study, confirming their functional link. C. nigriceps polydnavirus (CnPDV) plays a major role in the inactivation of pupally committed host prothoracic glands, while putative factors occurring in the host haemolymph do not seem to be of particular importance at that developmental stage. Southern blot hybridization indicates the occurrence of PKI(protein kinase inhibitor)-like genes in the C. nigriceps genome, which, in contrast, are undetectable in H. virescens. PMID- 12769880 TI - Mechanism of parasitism-induced elevation of haemolymph growth-blocking peptide levels in host insect larvae (Pseudaletia separata). AB - Growth-blocking peptide (GBP) has been purified for the first time from the haemolymph of the host armyworm Pseudaletia separata whose growth is inhibited and shows developmental arrest in the last larval instar stage when parasitized by the parasitoid wasp Cotesia kariyai. GBP naturally occurs in the haemolymph of lepidopteran larvae but its concentration is very low during the last larval instar in comparison with that in the penultimate larval instar. However, by 24h after parasitization or polydnavirus (PdV)-infection on day 0 of the last larval instar, a four-fold increase in GBP level, compared with synchronous non parasitized control larvae, is observed. Although Northern blot analysis indicates that GBP mRNA is transcribed in brain-nerve cord and fat body, plasma GBP is likely to be secreted mainly from fat body because the GBP mRNA level is approximately 100-fold higher in fat body than that in brain-nerve cord. RT-PCR analysis demonstrates the constant expression of GBP mRNA in both parasitized (or PdV-infected) and non-parasitized larval fat body, which suggests that parasitism does not influence transcriptional level, but might influence post transcriptional level to elevate plasma GBP concentration. This interpretation was supported by estimating GBP precursor levels in fat body of PdV-infected and non-infected larvae. Virus infection appears to elevate the GBP precursor levels in fat body to about six times greater than that in non-infected last instar larvae by 6h after PdV-injection. The GBP processing enzyme activity that occurs in Golgi body-rich extract of the fat body is increased by about 90% after parasitization or PdV-injection. PMID- 12769881 TI - Effect of DIMBOA on growth and digestive physiology of Sesamia nonagrioides (Lepidoptera: noctuidae) larvae. AB - The hydroxamic acid 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA) was assessed for its effect on growth and digestive physiology of larvae of the stalk corn borer Sesamia nonagrioides Lef. Nutritional indices and activities of some digestive and detoxification enzymes were determined for larvae feeding on a DIMBOA-containing diet for the first two days of the third instar (short-term feeding assays), and from neonates to third instar (long-term feeding assays). DIMBOA reduced the relative growth rate and the efficiency of conversion of ingested food without affecting the relative consumption rate in long-term feeding assays, but it had no effect in short-term assays. Moreover, elastase like activity was significantly increased by DIMBOA in short-term feeding assays, whereas microsomal oxidase activity was increased and esterase activity was reduced in long-term feeding assays. In vitro, DIMBOA inhibited the activities of carboxypeptidases, aminopeptidase, glutathione S-transferase and esterase, but it had no effect on trypsin, chymotrypsin and elastase. The implications of the altered levels of proteases and detoxification enzyme activities on the digestive physiology of larvae feeding on DIMBOA-containing diets are discussed. PMID- 12769882 TI - The Release of Isoprenoids by Locust Corpora Allata in vitro. AB - Corpora allata of the African locust Locusta migratoria, incubated in vitro, biosynthesized together with juvenile hormone III (JH-III), several molecules labelled by both [2-(14)C] sodium acetate and L-[methyl-(3)H] methionine. By a combination of chromatographic procedures including reverse phase-high performance liquid chromatography (HPLC), normal phase HPLC and thin layer chromatography (TLC), four labelled compounds were separated. They were isoprenoids, as revealed by inhibition of their synthesis by the hydroxy methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitor fluvastatin and restoration by exogenous mevalonolactone. They could be produced by incubating corpora allata with JH-III, suggesting that they were JH-III metabolites. They were produced by the corpora allata from both males and females and released into the incubation medium. Their rate of synthesis changed considerably depending on the sample, and in some cases they were the major isoprenoic products of the corpora allata. PMID- 12769883 TI - Colour Pattern Plasticity in the Hoverfly, Episyrphus balteatus: The Critical Immature Stage and Reaction Norm on Developmental Temperature. AB - Episyrphus balteatus is phenotypically plastic with respect to abdominal colour pattern. It was hypothesized that developmental temperature was the environmental cue governing this plasticity. The length of different immature stages was manipulated by altering the rearing temperatures. It was shown that all stages of the pupal developmental period were equally important in influencing adult phenotype. Lengthened pupal period, achieved by reducing rearing temperature, resulted in darker individuals. Adjusting the length of the larval period had no affect on adult colour pattern. Colour pattern was quantified using image analysis. Reaction norms of black pigment on each tergite against pupal developmental temperature were constructed. All reaction norms were approximately linear. Females were darker than males at comparable temperatures. In both sexes tergite 2 was the most plastic, whereas tergite 3 was the least plastic. The results are discussed in connection with the colour plasticity having a possible thermoregulatory function. PMID- 12769884 TI - Physiological trade-offs between reproduction, flight capability and longevity in a wing-dimorphic cricket, Modicogryllus confirmatus. AB - Physiological and morphological comparisons were made between the short-winged (SW) and long-winged (LW) morphs of a cricket, Modicogryllus confirmatus, to determine the cost of the flight capacity and the physiological mechanisms underlying trade-offs between different life history traits related to migration and reproduction. Both wingmorphs grew at a similar rate and no consistent correlation was found between nymphal development and adult body size. The metathoracic muscles at adult emergence represented 4.2% of the wet body weight in the SW morph and 10.5% in the LW morph. Fat content of the body at adult emergence was positively correlated to dry body weight, but no significant difference was found in mean fat content between the two wingmorphs or between sexes after body weight was adjusted. SW females fed ad libitum produced significantly more eggs than LW females during the first 20days of adult life. Egg production was not correlated to either body size or the duration of nymphal development. LW adults lived longer than SW ones when kept with water alone or when given various amounts of food on day 1 and otherwise kept with water alone. In females, a highly significant correlation was found between longevity and egg production, indicating the presence of a trade-off. LW females mainly allocated the energy from food to flight muscle development and general maintenance of the body rather than to egg production, whereas SW females used it for egg production and longevity. LW females that had been de-alated at adult emergence histolyzed the flight muscle and used the energy from food for egg production almost exclusively. These results suggest that energy allocation and trade-offs after adult emergence may play crucial roles in the functional differentiation of the two wingmorphs. PMID- 12769885 TI - The effects of aposymbiosis and of an infection with Blastocrithidia triatomae (Trypanosomatidae) on the tracheal system of the reduviid bugs Rhodnius prolixus and Triatoma infestans. AB - We have investigated the effects of the absence of symbionts and of infection with a trypanosome on the tracheal supply to different organs in larvae of the blood-sucking bugs, Rhodnius prolixus and Triatoma infestans. In bugs grown without symbionts there were extensive reductions in the tracheal supply to all the internal organs examined. These bugs also excreted less fluid after blood meals, perhaps because of a reduction in the oxygen supply to the Malpighian tubules. Inclusion of vitamin B in the diet of these affected insects reversed both the adverse effects on diuresis and on the extent of the respiratory supply to the different internal organs. The results of these studies suggest that vitamin B may play a key role in the development and maintenance of an adequate tracheal supply to the tissues in these insects. Infection with the trypanosome Blastocrithidia triatomae also greatly reduced the density of tracheoles supplying the rectum, small intestine and Malpighian tubules in infected T. infestans, but not in R. prolixus. It is possible, therefore, that the parasite exerts at least part of its pathogenic effect by causing a vitamin B deficiency, that in turn affects oxygen supply to the tissues. PMID- 12769887 TI - Feeding behavior of Ceratitis capitata (Diptera, Tephritidae): influence of carbohydrate ingestion. AB - Experiments were conducted on Ceratitis capitata larvae and adults to determine the influence of sugar (glucose and sucrose) ingestion on some aspects of the feeding behavior of this species. The results indicate that larval choice of a diet containing glucose or sucrose is not influenced by the rearing diet, by carbohydrate deprivation or by previous experience (induction). Carbohydrate deprivation did not alter the discrimination threshold of the larvae for sucrose. In adult females, the discrimination threshold for sucrose was unchanged when the flies were submitted to carbohydrate deprivation during the adult phase although ingestion by deprived females was 35% higher than ingestion by the control group. However, adults submitted to carbohydrate deprivation during the larval phase suffered a profound reduction in discrimination threshold for sucrose. PMID- 12769886 TI - In vivo Responses of Honey Bee Midgut Proteases to Two Protease Inhibitors from Potato. AB - Potato protease inhibitors, POT-1 and POT-2, were fed to newly emerged adult honey bees in cages at different doses in either sugar syrup (0.2 or 0.01% w:v) or pollen food (1 or 0.2% w:w). In vivo activities of three digestive endopeptidases (trypsin, chymotrypsin and elastase) and one exopeptidase (leucine aminopeptidase; LAP) were measured after 3 or 8days' exposure of bees to inhibitor. Enzyme activities were significantly lower at day 8 than at day 3, except for elastase, which did not change. POT-2 significantly reduced the activity of all endopeptidases at both timepoints, regardless of the dose level or the medium in which the inhibitor was administered. POT-1 acted in a similar manner, except that 0.01% POT-1 in syrup had no effect on bees. There was no consistent trend in changes in LAP activity. Bees fed either inhibitor at 1% in pollen or at 0.2% in syrup had significantly reduced lifespans, with the effect of the pollen treatment being greater than the syrup treatment. The survival of bees fed POT-1 or POT-2 at 0.2% in pollen or 0.01% in syrup did not differ from the controls. PMID- 12769888 TI - The influence of bacterial species and intensity of infections on nodule formation in insects. AB - Nodulation is the predominant cellular immune reaction to bacterial infection in insects. Nodulation is a complex process involving an unknown number of discrete cellular actions. Currently, there is only limited information on the signal transduction mechanisms that result in nodulation. In older larvae of the tobacco hornworm, Manduca sexta, and of the tenebrionid beetle, Zophobas atratus, eicosanoids are involved in one or more steps in the overall process, and treating these insects with inhibitors of eicosanoid biosynthesis prior to bacterial infection severely impairs their ability to form nodules. In this paper we address more detailed questions on eicosanoid-mediated nodulation. The nodulation reaction to bacterial infection occurs in all larval stages we examined, specifically, second, third, and fourth instars of M. sexta. In both species, the number of nodules formed in response to bacterial infection is related in an exponential way to the number of bacterial cells in the infection. Nodulation is also not related to larval size. We also found that nodulation intensity varies according to the species of infecting bacteria. PMID- 12769889 TI - Physiology of drought tolerance and cold hardiness of the Mediterranean tiger moth Cymbalophora pudica during summer diapause. AB - Prepupae of the arctiid moth Cymbalophora pudica spend spring and summer months in a summer diapause (aestivation), the duration of which is photoperiodically controlled. Cold hardiness, drought tolerance and some physiological and biochemical parameters were measured in aestivating prepupae. Large amounts of metabolic reserves, in the form of lipids and glycogen, accumulated prior to aestivation. Glycogen served as the main metabolic fuel for aestivating prepupae. Metabolic rate decreased rapidly after the onset of the inactive prepupal stage and remained low (5-15% of the level in active larva) during aestivation. A spontaneous increase of the respiration rate occurred before pupation. Neither low mol. wt sugars or alcohols (polyols) accumulated nor the haemolymph osmotic pressure changed during aestivation. Drought tolerance of aestivating prepupae was high (no decrease in survival after exposure to r.h.<10% at a temperature of 23 degrees C for a substantial part of diapause) owing to their extensive capacity to stabilize the relative body water content irrespective of the r.h. of surrounding air. Cold hardiness was low (>90% decrease in survival after exposure to -7 degrees C for 24h). Cold and drought acclimations did not lead to significant changes in the measured physiological and biochemical parameters but cold (not drought) acclimation caused a significant increase in cold hardiness. Neither drought tolerance nor the increase in cold hardiness after cold acclimation appear to be related to presence/accumulation of polyols in aestivating C. pudica prepupae. PMID- 12769890 TI - Phosphorylation polymorphism in the yolk protein 2 of Drosophila hawaiiensis. AB - An intraspecific polymorphism in the electrophoretic migration pattern of the yolk proteins in D. hawaiiensis was established and characterized. The polymorphism includes yolk protein migration patterns of two, three, or four bands by polyacrylamide gel electrophoresis. Peptide mapping analysis demonstrates that in the two band migration pattern YP2 comigrates with YP3, whereas in the three and four band migration patterns YP2 migrates between YP1 and YP3 in addition to comigrating with YP3. It further demonstrates that the top two bands of the four band migration pattern consists of YP1. Phosphatase treatment of the yolk proteins establishes that the different electrophoretic migration patterns of YP2 are caused by different degrees of phosphorylation. It is suggested that the YP1 polymorphism is caused by a yp1 gene modification and that the YP2 polymorphism is caused by two different post-translational processing paths. PMID- 12769891 TI - Receptor neuron responses to potential sex pheromone components in the caddisfly Rhyacophila nubila (Trichoptera: Rhyacophilidae). AB - Olfactory single sensillum responses of male and female caddisflies Rhyacophila nubila (Trichoptera: Rhyacophilidae) were examined with the tungsten penetration technique to investigate the presence of receptor neurons tuned to potential sex pheromone components. The compounds tested in the experiment were heptan-2-one, nonan-2-one, (R)-heptan-2-ol, and (R)-nonan-2-ol, which have all been found to be present in female abdominal glands, plus the two antipodes (S)-heptan-2-ol and (S)-nonan-2-ol. Four types of receptor neurons were found in equal proportions in males and females. One type responded primarily to heptan-2-one. A second type responded primarily to (R)-heptan-2-ol and a third type primarily to its enantiomer (S)-heptan-2-ol. The fourth type of receptor neuron responded to nonan 2-one, (R)-nonan-2-one, and (S)-nonan-2-ol, with a bias to nonan-2-one. The receptor neurons responding to the tested compounds made up a great majority of the contacted neurons in both males and females. PMID- 12769892 TI - Mandibular Gland Volatiles and Their Ontogenetic Patterns in Queen Honey Bees, Apis mellifera carnica. AB - Extracts of mandibular glands taken from adult queens of the honey bee, Apis mellifera carnica, were analysed by gas chromatography-mass spectroscopy. More than 100 compounds could be identified among which oxygenated fatty acids with six, eight, 10 and 12 carbon atoms are particularly interesting since they show structural relationships to the queen substance, (E)-9-oxo-2-decenoic acid. Changes in the patterns of volatiles were followed up from emergence until the full dominant status of an egg-laying queen in a strong colony. Generally, the amount of volatiles per gland was found to increase with age. The final level of queen substance (9-ODA) content is reached at the postmating stage about 10 days after emergence. Ontogenetic patterns of concentrations were determined for those components regarded to predominantly contribute to the royal pheromone. Characteristic compositions of signals, possibly involved in the premating, mating and postmating dominance status of a honey bee queen are discussed. Copyright 1997 Elsevier Science Ltd. All rights reserved PMID- 12769894 TI - Orientation-sensitive Neurons in the Brain of the Honey Bee (Apis mellifera). AB - Recent behavioural experiments have shown that bees are able to distinguish vertically presented patterns with orientation cues, although the locations of areas of black are randomized. To discriminate between two orientations, the bees must possess more than one orientation-sensitive neuron type. Therefore, the aim is to search for different types of orientation-sensitive cells of the honey bee, and measure their receptive field, velocity sensitivity and contrast sensitivity. Orientation-sensitive cells with two different types of orientation tuning-curves were recorded intracellularly in the mid-brain of the honey bee when the stimulus was a narrow bar (bar width = 5 degrees ). These cells are sensitive to bar movement within their large receptive field, which covers the visual field of one eye. They are quite distinct from the well-known directional motion detectors. The contrast sensitivity of the orientation-sensitive cells recorded in this study corresponds to results from behavioural experiments. The velocity sensitivity curves of the orientation-sensitive cells differ from those of the direction-sensitive cells. Measurements of orientation sensitivity and contrast sensitivity when the stimulus is a wide bar (bar width = 10 degrees ), done in different eye regions, suggest that each orientation-sensitive cell receives visual signals from an array of orientational subunits within its receptive field. The correspondence between these physiological results and the results of recent behavioural experiments are discussed. Copyright 1997 Elsevier Science Ltd. All rights reserved PMID- 12769893 TI - The Effects of Ingested 20-Hydroxyecdysone on the Larvae of Aglais urticae, Inachis io, Cynthia cardui (Lepidoptera: Nymphalidae) and Tyria jacobaeae (Lepidoptera: Arctiidae). AB - A comparative survey was carried out to investigate the effects, distribution and metabolism of ingested 20-hydroxyecdysone in four species of lepidopteran larvae in relation to the phytoecdysteroid content of the insect's host plants. Analysis of the leaves of the host plants of each of the species revealed a strong relationship between the levels of phytoecdysteroids and the relative tolerance of the larvae to ingested 20-hydroxyecdysone. Monophagous or oligophagous species (Aglais urticae, Inachis io) feeding on ecdysteroid-negative host plants were either deterred from feeding or showed marked abnormalities in growth and development after incorporation of 20-hydroxyecdysone in their diets. Oligophagous or polyphagous species (Tyria jacobaeae, Cynthia cardui) which feed on host plants from families which are known to contain phytoecdysteroid-positive species, were able to tolerate low levels of 20-hydroxyecdysone in their diets, but exhibited developmental defects at high concentrations. These species were termed semi-tolerant. In each of the species, ingested [3H]20-hydroxyecdysone appeared to follow the same fate as injected [3H]20-hydroxyecdysone. The data are compared to those obtained in previous studies, where truly polyphagous species were shown to tolerate very high concentrations of 20-hydroxyecdysone in their diets by the production of ecdysteroid 22-fatty acyl esters. Copyright 1997 Elsevier Science Ltd. All rights reserved PMID- 12769895 TI - The Interplay between Patency, Microsomal Na(+)/K(+) ATPase Activity and Juvenile Hormone, in Tenebrio molitor Parasitized by Hymenolepsis diminuta. AB - Beetles infected with metacestodes of the rat tapeworm, Hymenolepis diminuta, exhibit reduced fecundity, due to alterations in vitellogenesis. Follicle cell patency is retarded and inefficient vitellogenin uptake ensues. Here, we have reassessed patency and its stimulation by JH III at day 3 post-infection, when the most detrimental changes are observed in other ovarian processes. In Rhodnius prolixus, patency is believed to be brought about by the action of a JH-dependent membrane-bound Na(+)/K(+) ATPase (EC 3.6.1.3); however, this had not been established in Tenebrio molitor. Therefore, the properties of the enzyme, with respect to optimal assay conditions and juvenile hormone dependency, are reported. Maximal stimulation occurred between 50 and 500 nM JH III, a range over which greatest increases in patency were also observed. In infected insects, a 35% reduction in Na(+)/K(+) ATPase activity was noted, but exposure to 50 nM JH III is sufficient for stimulation to a specific activity 89% that of JH-treated controls. In a similar fashion, patency in infected insects is reduced, but can be 'rescued' by 50 nM JH III. Moreover, in the absence of exogenous hormone, patency in infected beetles can be elevated to control levels after in vitro culture (6 h), with exchange of medium every 2 h. The possibility that such reversible decreases in enzyme activity and patency are caused by a JH binding inhibitor molecule is discussed. Copyright 1997 Elsevier Science Ltd. All rights reserved PMID- 12769896 TI - Characterization of a Monoclonal Antibody Against a 180 kDa Hemocyte Polypeptide Involved in Cellular Defence Reactions of the Stick Insect Bacillus rossius. AB - Defence properties of hemoctyes were investigated using the anti-hemocyte monoclonal antibody BrH1 obtained by immunizing mice with 2% paraformaldehyde fixed hemoctyes of the stick insect Bacillus rossius. In Western blot analysis, the antibody recognized a 180 kDa antigen in hemocyte cell lysates, whereas fat body lysates and cell-free hemolymph were negative. In immunofluorescence analysis of cultured or freshly collected hemoctyes, BrH1 stained intracellular antigen(s) in detergent-treated cells. Transverse cryosections of adult stick insects probed by immunofluorescence with BrH1 showed in situ the scattered distribution of hemoctyes inside the haemocoel. The antigen(s) recognized by BrH1 appears to be involved in cell defence hemocyte-mediated mechanisms, as evidenced by the fact that cryosections of insects challenged in vivo with yeast cells, bacteria, or polystyrene latex particles and probed with BrH1 showed an accumulation of antigen surrounding the injected stimuli. Copyright 1997 Elsevier Science Ltd. All rights reserved PMID- 12769897 TI - A Peptide from the Male Accessory Gland in Leptinotarsa decemlineata: Purification, Characterization and Molecular Cloning. AB - Our interest in the male accessory glands (MAGs) of Leptinotarsa decemlineata was raised recently by our finding that certain cells produce a secretory substance that is recognized by one of our monoclonal antibodies (MAC-18), developed for the immunohistochemical demonstration of peptidergic neurons in the brain. We undertook to isolate this substance, presumably a peptide, to find out more about its role in the post-mating physiology of the recipient of this peptide, the mated female. This paper describes the purification and chemical characterization of the immunoreactive peptide from 100 pairs of male accessory glands. The peptide was purified by two subsequent reversed-phase-HPLC runs, and fractions were analyzed on Western blots that were immunostained by MAC-18. This indicated the presence of an 8 kDa peptide in the MAG. Partial analysis of the N-terminal amino acids by automated Edman degradation revealed a sequence of 40 amino acid residues. To obtain the full amino acid sequence of this peptide, the technique of reverse transcriptase PCR (3'RACE) was used. A PCR product of 350 bp was obtained, which encoded the 3'-end of the mRNA. After cloning and sequencing, this product contained most of the genetic information of the MAG peptide. The PCR product was also used as a probe for screening a cDNA library constructed from mRNA extracted from MAGs. The nucleotide sequence coding for the signal peptide was elucidated by 5'RACE. The cDNA and 5'RACE clones were analyzed and sequenced. The sequence of the cDNA clone contained an insert of 411 bp, which agreed well with the mRNA size measured by Northern blotting. Translation of the DNA sequences confirmed the data from partial amino acid sequence analyses and also predicted the remainder of the amino acid sequence. The entire peptide, designated Led-MAGP, consists of 74 residues; its mass was calculated and confirmed by mass spectrometry at 7971 Da. The peptide contains seven imperfect hexa-repeats, and this hexa-repeat sequence shows remarkable similarity to the hexa-repeat section of the chicken prion protein. The physiological function of the peptide has yet to be determined, but the hexa-repeat motif has recently been identified as the signal that induces internalization of the prion protein by coated-pit mediated endocytosis. Possible implications for the control of reproductive activities in L. decemlineata are discussed. Copyright 1997 Elsevier Science Ltd. All rights reserved PMID- 12769898 TI - Influence of Pyriproxyfen on the Expression of Haemolymph Protein Genes in the Colorado Potato Beetle, Leptinotarsa decemlineata. AB - Pyriproxyfen, a potent juvenile hormone analogue for the Colorado potato beetle, Leptinotarsa decemlineata, was applied topically to last-instar larvae and short day adults at different times after moulting. The effect of the hormone analogue on concentration and composition of protein in the haemolymph was studied at different intervals after pyriproxyfen application. The hormone analogue had little effect on total protein concentration of the haemolymph, but affected protein composition. Diapause protein 1 was prevented from being synthesized if pyriproxyfen was applied before the gene was activated and disappeared from the haemolymph if applied after the gene had been expressed. It therefore inactivated the gene for diapause protein in both larvae and adults. Pyriproxyfen also induced appearance of vitellogenin at both stages, indicating induction of expression of the vitellogenin gene. It also affected the stability of mRNA for diapause protein. The analogue caused mRNA for diapause protein 1 to disappear untimely compared to controls in last-instar larvae and short-day adults. The response of adults to the JHA was much more pronounced than that of larvae, although the analogue had a strong biological effect on last-instar larvae because it prevented metamorphosis at low doses. Copyright 1997 Elsevier Science Ltd. All rights reserved PMID- 12769899 TI - Caste Determination in Bombus terrestris: Differences in Development and Rates of JH Biosynthesis between Queen and Worker Larvae. AB - To study the possible role of juvenile hormone in caste determination in Bombus terrestris, we measured development and rates of juvenile hormone biosynthesis in vitro in larvae destined to develop into either workers or queens. Larvae of both castes developed through four instars and had the same growth rates. However, the duration of the instars was longer for queen larvae, and their head width at the third and fourth instars was significantly larger. After validating the well known radiochemical assay of JH for bumble bee larvae, we show that worker larvae corpora allata exhibited a constant and low rate of JH biosynthesis, never more than 5 pmol JH/h/pair. Queen larvae, in contrast, had two peaks of JH biosynthesis: a small one during the first instar, which has previously been correlated with caste determination; and a large peak, previously undetected, above 40 pmol JH/h/pair, during the second and third instars. We suggest that caste determination in this species is mediated by JH and that the duration of larval instars is a key factor. The possibility that the queen influences caste determination via an effect on instar duration is also discussed. Copyright 1997 Elsevier Science Ltd. All rights reserved PMID- 12769900 TI - Isolated Apolipophorin III from Galleria mellonella Stimulates the Immune Reactions of This Insect. AB - Apolipophorin III (apoLp-III) was isolated from the haemolymph of last instar larvae of Galleria mellonella. The ultraviolet (u.v.) spectrum and the N-terminal amino acid sequence reveal high similarities with the apoLp-III from Manduca sexta. The protein is heat-stable. The molecular mass of apoLp-III was determined to be 18 077 Da using mass spectrometry. The heat treatment (90 degrees C, 30 min) resulted in a pI shift from 6.6 for the non-heated to 6.1 for the heat treated apoLp-III without change in the molecular mass, indicating that a conformational change might have been caused by the heat treatment, rather than covalent alterations. Intrahaemocoelic injection of pure apoLp-III into last instar G. mellonella larvae is followed by a dose-dependent increase of antibacterial activity in cell-free haemolymph of treated larvae 24 h after injection. Furthermore, pure apoLp-III enhances the phagocytic activity of isolated haemocytes in vitro. The newly discovered role of apoLp-III in inducing immune-related functions in insects is discussed in regard to the known features of this molecule in lipid metabolism. Arylphorin, another heat-stable protein in G. mellonella haemolymph, was likewise isolated in this study. The protein was identified by N-terminal protein sequencing, the sequence obtained exactly matches the known sequence data for this protein. Copyright 1997 Elsevier Science Ltd. All rights reserved PMID- 12769901 TI - Comparing Different Measures of Heat Resistance in Selected Lines of Drosophila melanogaster. AB - Lines of the fly Drosophila melanogaster were selected for increased knockdown resistance to heat (39 degrees C) in a long tube. One set of lines was selected following prior heat hardening (1 h at 37 degrees C) and another without hardening. Each set consisted of three replicate selection lines and three unselected controls. Lines were tested for correlated responses to selection, in order to define the nature of knockdown resistance. Selection had a large effect on knockdown resistance, but selected lines did not differ from controls for knockdown time in small vials, survival, or recovery time following exposure to heat. Selection with and without hardening influenced the hardening response in the long tube, but not in small vial assays of resistance. The hardened selection lines had decreased resistance to ethanol and a reduced dry weight, whereas the non-hardened lines did not show these changes. The same correlated responses were also evident in two generation experiments on unselected flies. Both sets of lines showed a reduction in activity when tested at 37 degrees C, but not at 25 degrees C. These results indicate that different measures of heat resistance are surprisingly unrelated, and suggest that subtle features of the selective environment influence responses and correlated responses to selection. Copyright 1997 Elsevier Science Ltd. All rights reserved PMID- 12769902 TI - Parasitism-induced Effects on Host Growth and Metabolic Efficiency in Tobacco Hornworm Larvae Parasitized by Cotesia congregata. AB - Parasitism by the braconid wasp Cotesia congregata affects the growth of Manduca sexta larvae in a parasitoid 'dose-dependent' fashion. Following parasitization of fourth-instar larvae, more heavily parasitized larvae grew larger compared to those containing fewer parasitoids due to an increase in host dry weight. The differences in host mass appeared to arise after oviposition. A 'dose-dependent' enhancement of host dry weight would appear nutritionally beneficial for the parasitoids developing in more 'crowded' hosts. The efficiencies of conversion of ingested and digested food to body mass and the approximate digestibility of the diet ingested by the host caterpillar did not vary significantly with clutch size although parasitoids took slightly longer to develop in the more heavily parasitized hosts. Larval parasitoids developing in the presence of many competitors weighed up to 50% less than those developing in hosts with fewer endoparasitoids, although the weight of adult female parasitoids did not vary significantly with wasp clutch size. The maximum number of emerging wasps was 200 parasitoids, possibly representing the host's 'carrying capacity' for larvae parasitized in the fourth-instar. The ratio of emerging to non-emerging parasitoids decreased as parasitoid clutch size increased, with few or none emerging from very heavily parasitized hosts containing more than 400 parasitoids. Copyright 1997 Elsevier Science Ltd. All right reserved PMID- 12769903 TI - Behavioral and Electrophysiologic Responses of Drosophila melanogaster to Prolonged Periods of Anoxia. AB - Sensitivity to anoxia varies tremendously among phyla and species. Most mammals are exquisitely sensitive to low concentrations of inspired oxygen, while some fish, turtles and crustacea are very resistant. To determine the basis of anoxia tolerance, it would be useful to utilize a model system which can yield mechanistic answers. We studied the fruit fly, Drosophila melanogaster, to determine its anoxia resistance since this organism has been previously studied using a variety of approaches and has proven to be very useful in a number of areas of biology. Flies were exposed to anoxia for periods of 5-240 min, and, after 1-2 min in anoxia, Drosophila lost coordination, fell down, and became motionless. However, they tolerated a complete nitrogen atmosphere for up to 4 h following which they recovered. In addition, a nonlinear relation existed between time spent in anoxia and time to recovery. Extracellular recordings from flight muscles in response to giant fiber stimulation revealed complete recovery of muscle-evoked response, a response that was totally absent during anoxia. Mean O(2) consumption per gram of tissue was substantially reduced in low O(2) concentrations (20% of control). We conclude from these studies that: (1) Drosophila melanogaster is very resistant to anoxia and can be useful in the study of mechanisms of anoxia tolerance; and (2) the profound decline in metabolic rate during periods of low environmental O(2) levels contributes to the survival of Drosophila. Copyright 1997 Elsevier Science Ltd. All rights reserved PMID- 12769904 TI - Role of the Corpus Allatum in the Control of Adult Diapause in the Blow Fly, Protophormia terraenovae. AB - Adults of the blow fly, Protophormia terraenovae developed their ovaries promptly after emergence under long-day conditions at 25 degrees C, although they entered diapause under short-day conditions at 20 degrees C. Application of a juvenile hormone analog (JHA), implantation of the corpus allatum (CA) from reproductive adults, or transection of the nervus corporis allati induced ovarian development under diapause-inducing conditions. Removal of the CA suppressed ovarian development under diapause-averting conditions, and JHA application induced ovarian development in allatectomized adults. Therefore adult diapause in P. terraenovae is caused by a reduction of juvenile hormone secretion from the CA, and the endocrine activity of the CA is inhibited through a nervous pathway by the brain in diapause adults. Copyright 1997 Elsevier Science Ltd. All rights reserved PMID- 12769905 TI - Glycine in Digestive Juice: a Strategy of Herbivorous Insects Against Chemical Defense of Host Plants. AB - A large amount of free glycine was found in the digestive juice of several Lepidoptera larvae and appears to be secreted. The amount of glycine differs greatly among species and seems to correlate with the host plant. Several species of Lepidoptera from different famileis which feed on the privet tree, Ligustrum obtusifolium, show extremely large amounts of free glycine. The concentration of glycine reached 57 &mgr;moles/g (0.4%). The leaves of the privet tree have an extremely strong protein denaturing activity, enough to make protein completely non-nutritive. This denaturing property closely resembles the activity of polyphenolics, digestion inhibitory compounds common among plants. In vitro studies showed that free glycine completely inhibits the denaturing activity of the privet leaves. These results suggest that insects secrete glycine to counter the chemical defenses of host plants. Copyright 1997 Published by Elsevier Science Ltd. All rights reserved PMID- 12769906 TI - The Effects of G Protein Modulators on the Labellar Taste Receptor Cells of the Fleshfly (Boettcherisca peregrina). AB - A nonhydrolyzable G protein activator (guanosine 5'-O-(3-thiotriphosphate); GTPgammaS) and a G protein inhibitor (guanosine 5'-O-(2-thiodiphosphate); GDPbetaS) were introduced into the labellar taste receptor cells of the fleshfly by treatment of their receptive membranes beneath the tip opening of the chemosensory hair with each reagent in 0.03% deoxycholate solution for 4 min. After treatment with GTPgammaS, the responses of the sugar receptor cell to D glucose, D-fructose, L-phenylalanine and L-valine and that of the salt receptor cell to cyclic AMP were markedly enhanced, compared with those after treatment with deoxycholate alone. Treatment with GDPbetaS depressed these responses. These results strongly suggest that the responses are mediated by G protein. However, the response of the salt receptor cell to NaCl was not affected by treatment with either GTPgammaS or GDPbetaS, and thus the response to NaCl clearly is not elicited through a G protein-regulated mechanism. Copyright 1997 Elsevier Science Ltd. All rights reserved PMID- 12769907 TI - Cellulose and Xylan Utilisation in the Lower Termite Reticulitermes speratus. AB - The distribution of the enzymes of cellulose and xylan metabolism namely endo beta-1,4-glucanase, beta-glucosidase, endo-beta-1,4-xylanase and beta-xylosidase activities, in Reticulitermes speratus (Kolbe) was measured both in the salivary glands and in the major gut sections and along the length of the gut in freshly collected termites. The majority of the endo-beta-1,4-glucanase activity (77.8%) was found in the salivary glands which also contained 23.9% of the beta glucosidase activity. At least 70% of the remaining activity was located in the anterior section of the hindgut. A small amount of endo-beta-1,4-xylanase activity (2.4%), but no beta-xylosidase activity, was present in the salivary glands. The majority of these activities were in the anterior section of the hindgut. The RQ of freshly collected termites at 25 degrees C was 1.03+/-0.01. Maintaining termites for 16 days on wood, cellulose and xylan showed that the RQ values of termites fed on wood or xylan were not significantly different from those of freshly collected termites but significantly increased when maintained on cellulose. The RQ of starved termites after 11 days was 0.81+/-0.02. There were three effects on protozoan populations of feeding termites xylan for 20 days. One species, Dinenympha parva was not affected, while five others, Pyrsonympha grandis, Holomastigotes elongatum, Dinenympha rugosa, Dinenympha leidy and Dinenympha porteri survived for 20 days but slowly decreased in numbers. The numbers of P. grandis and D. leidy surviving for 20 days were significantly different from those in starved termites. The third group comprising the two large species, Teratonympha mirabilis and Trichonympha agilis and three small species, Pyrsonympha modesta, Dinenympha exilis and Dinenympha nobilis disappeared within 15 days as in starved termites. It is suggested that protozoa in the first two groups are xylanolytic. Protozoan populations on wood and cellulose diets were not markedly affected. Selective removal of the protozoa by u.v. irradiation led to the loss of xylanolytic activity and a life span comparable to starved termites. Copyright 1997 Elsevier Science Ltd. All rights reserved PMID- 12769908 TI - Permeability of the Peritrophic Envelopes of Herbivorous Insects to Dextran Sulfate: a Test of the Polyanion Exclusion Hypothesis. AB - We tested the hypothesis that the permeability of the peritrophic envelope in herbivorous insects is greatly reduced for polyanions as a result of an extensive network of anionic sites in the proteoglycans of the matrix. 14C-Dextran sulfate (polyanionic, 8000 M(w)) and fluorescein isothiocyanate-labeled (FITC) dextran (monoanionic, 9400 M(w)) were introduced together into the endoperitrophic space of the midguts of Orgyia leucostigma (Lepidoptera) larvae and Melanoplus sanguinipes (Orthoptera) adults. In all cases more of the 14C-dextran sulfate permeated the peritrophic envelope than the FITC-dextran, the opposite of the result predicted by the polyanion exclusion hypothesis. We conclude that polyanion exclusion is not a mechanism that contributes significantly to the permeability properties of the peritrophic envelopes of these two species, or that explains the failure of tannic acid to cross the peritrophic envelopes of lepidopteran larvae. Copyright 1997 Elsevier Science Ltd. All rights reserved PMID- 12769909 TI - Effects of Temperature on Weak Circadian Eclosion Rhythmicity in Chymomyza costata (Diptera: Drosophilidae). AB - The adult eclosion rhythms of two Japanese strains of Chymomyza costata were studied in diel thermoperiods of different duration and at different average temperatures. One of the strains had the 'wild-type' photoperiodic larval diapause and the other was a mutant strain lacking the photoperiodic response. At constant temperatures the wild-type strain had weakly rhythmic eclosion in diel photoperiods while the mutant strain was arrhythmic. Free-running rhythms could scarcely be observed at all. The amplitude of the rhythm of both strains was much higher in diel temperature cycles than in corresponding light-dark cycles, and generally higher in continuous darkness than in continuous light. When the average temperature under entraining conditions was lowered, the rhythmicity increased and the median of the eclosion peak was displaced to later hours in the entraining cycle. Both strains were rhythmic at the lowest temperatures, i.e. near 10 degrees C. At low temperatures the majority of the eclosions occurred during the high temperature phase or light phase of the entraining cycle. Although the rhythm started well in the entraining temperature cycles, the subsequent free-running rhythm in constant conditions lasted for only 2-3 days. We concluded that the exceptionally weak rhythmicity of eclosions and the relative importance of temperature cues are adaptive traits which make it possible for this northern species to respond directly to favourable but unpredictable changes in its environment. Copyright 1997 Published by Elsevier Science Ltd. All rights reserved PMID- 12769910 TI - Physiological Mechanisms of Pheromonostatic Responses: effects of Adrenergic Agonists and Antagonists on Moth (Helicoverpa armigera) Pheromone Biosynthesis. AB - The adrenergic agonists octopamine, tyramine and clonidine inhibited the normal pheromonotropic action due to PBAN (pheromone biosynthesis activating neuropeptide) in incubations of intersegmental tissues that are situated between the 8th and 9th abdominal segments of the moth ovipositor tip. This inhibition was reversed in the presence of the adrenergic antagonists phentolamine, yohimbine and chlorpromazine. Incubations of 8th segments alone, which do not produce pheromone, resulted in elevated levels of intracellular cAMP in the presence of octopamine. The physiological significance of this phenomenon is unclear. However, clonidine (an alpha(2) selective agonist) did not duplicate octopamine stimulation of intracellular cAMP in 8th segment cultures. In intersegmental membrane cultures clonidine successfully duplicated the octopamine inhibition of both pheromone and intracellular cAMP production. The physiological significance of octopaminergic receptors mediating the inhibitory response of intersegments was investigated by experiments in vivo. When PBAN was injected into photophase females the normal pheromonotropic activity due to the injected PBAN dropped after 2h. In the presence of clonidine, normal peak stimulatory levels were never attained and a faster decline was observed. Clonidine also inhibited the pheromonotropic response of 24h-decapitated females to PBAN. Adrenergic antagonists successfully reversed the inhibitory effect of clonidine in decapitated females, but did not reverse the effect of clonidine in photophase females. In addition, when clonidine was injected into female moths during the scotophase normal peak pheromone titers were reduced although no effect on calling behavior was observed. Copyright 1997 Elsevier Science Ltd. All rights reserved PMID- 12769911 TI - Evolutionary Relationship between Diapause and Cold Hardiness in the House Spider, Achaearanea tepidariorum (Araneae: Theridiidae). AB - The relationship between diapause and cold hardiness of the house spider, Achaearanea tepidariorum, differed geographically. In a cool-temperate population, enhanced chilling tolerance and supercooling ability were observed in diapause individuals, whereas a subtropical population showed only chilling tolerance. Because this spider is considered to be of tropical origin, it would follow that the ancestral diapause of this spider was equipped with chilling tolerance, but not with an increased supercooling ability. It seems that the ability to lower the supercooling point evolved through natural selection in the course of expansion of this species to the northern climates. Copyright 1997 Elsevier Science Ltd. All rights reserved PMID- 12769912 TI - Charge Dependent Distribution of Endogenous Proteins within Vitellogenic Ovarian Follicles of Actias luna. AB - In vitellogenic ovarian follicles of Actis luna, internal Ca(2+) activity currents create an electrical gradient which influences the distribution of charged macromolecules between nurse cells and oocyte. We show that, between oocyte and nurse cells, there is an ionic gradient of 1-12 mV with the nurse cells being more electronegative than the oocyte by an average 3.5+/-0.2 mV(s.e.)(p<0.001). As previously reported for another saturniid, Hyalophora cecropia, the transbridge ionic gradient of luna: (1) is focused across the intercellular bridges, (2) is abolished by 200 &mgr;M vanadate and (3) includes a [Ca(2+)](i) gradient. Endogenous soluble proteins collected from control and from vanadate treated populations of nurse cells and oocytes were separated by two dimensional (2-D) gel electrophoresis and visualized with sliver stain. Densitometric analysis showed that 14 out of the 19 acidic proteins and six of the eight basic proteins studied, changed their oocyte-to-nurse cell distribution in consort with change in the transbridge ionic gradient. This suggests that a transbridge ionic gradient may be, at least within the saturniidae, a method for maintaining different molecular concentrations in nurse cells compared to oocytes. Copyright 1997 Elsevier Science Ltd. All rights reserved PMID- 12769913 TI - A Role for Ecdysteroids in the Induction and Maintenance of the Pharate First Instar Diapause of the Gypsy Moth, Lymantria dispar. AB - Several lines of evidence suggest a novel regulatory mechanism for diapause regulation in the gypsy moth. We propose that ecdysteroids play a role in the induction and maintenance of the pharate first instar larval diapause in this species. A 55 kDa gut protein that is indicative of diapause is expressed in intact and neck-ligated pharate larvae but is not expressed when a ligature is placed posterior to the prothorax, site of the prothoracic gland. Guts cultured in vitro for 12 h cease to synthesize the 55 kDa protein, but synthesis of the protein resumes if the culture medium is enriched with a prothorax extract from pharate larvae or a prothoracic gland extract from fifth instar larvae. Injection of 20-hydroxyecdysone or the ecdysteroid agonist, RH-5992, into isolated abdomens stimulates synthesis of the diapause-specific 55 kDa protein, suggesting that the essential factor from the prothorax is an ecdysteroid. KK-42, an imidazole derivative known to inhibit ecdysteroid biosynthesis, averts diapause when applied to prediapausing pharate first instar larvae, but this effect can be countered by application of 20-hydroxyecdysone or RH-5992, i.e. KK-42 treated pharate larvae that are exposed to an ecdysteroid or RH-5992 readily enter diapause. A chilling period (120 days at 5 degrees C) is normally adequate to prompt an immediate termination of diapause when pharate larvae are transferred to 25 degrees C, but if such larvae are held in hanging drop cultures with ecdysteroids they fail to terminate diapause. Together, these results suggest that ecdysteroids are essential for the induction and maintenance of diapause and imply that a drop in the ecdysteroid titer is essential for diapause termination. Copyright 1997 Elsevier Science Ltd. All rights reserved PMID- 12769914 TI - Light Cycles Given During Development Affect Freerunning Period of Circadian Locomotor Rhythm of period Mutants in Drosophila melanogaster. AB - We reared wild type (Canton-S) and period mutant flies, i.e., per(S) and per(L), of Drosophila melanogaster in constant darkness, constant light or 24h light dark cycles with various light to dark ratios throughout the development from embryo to early adult. The locomotor activity rhythms of newly eclosed individuals were subsequently monitored in the lighting conditions, in which they had been reared, for several days and then in constant darkness. Circadian rhythms were clearly exhibited in constant darkness even in flies reared in constant light and constant darkness as well as flies reared in light-dark cycles, but the freerunning period differed among groups. The results suggest that the circadian clock is assembled without any cyclical photic information, and that the light influences the developing circadian clock of Drosophila to alter the freerunning period. The effects of light on the rhythm differed in some aspects between per(L) flies and the other two strains. Possible mechanisms through which light affects the developing circadian clock are discussed. Copyright 1997 Elsevier Science Ltd. All rights reserved PMID- 12769915 TI - Processing of tactile information in neuronal networks controlling leg movements of the Locust. AB - The successful, coordinated, posture and locomotion of any animal requires a precise and continuous adjustment of limb movements by sensory feedback from extero- and proprioceptors associated with the legs. We here review the recent advances in our understanding of how specific local adjustments of the hind legs of the desert locust, Schistocerca gregaria, are made in response to tactile signals from two different classes of exteroceptor on a leg. The aim is to understand particular features of the organization of neuronal networks and how different types of constituent interneurones contribute to the processing of sensory signals. This information can then be used to define the design principles that govern the organization of sensory-motor networks. PMID- 12769917 TI - A hormone from the uterus of the tsetse fly, Glossina morsitans, stimulates parturition and abortion. AB - Unlike most insects, the tsetse female gives birth to a single, fully grown larva at the culmination of each pregnancy cycle. The expulsion of the larva is regulated by a hormone present in rich abundance within the female's uterus. The hormone elicits parturition when injected into neck-ligated females at late stages of pregnancy and abortion when injected at earlier stages. We refer to this highly active material (0.043 uterus equivalents stimulates parturition in 50% of the females) as parturition hormone. Injection of the active extract, which appears to be a peptide or small protein, initiates the series of blood pressure pulsations and uterine contractions normally associated with parturition. The discovery that a uterus extract from the flesh fly also elicits parturition in tsetse suggests that this hormone may be widely distributed in insects. PMID- 12769918 TI - Effects of Parasitism by the Braconid Wasp Cotesia congregata on Metabolic Rate in Host Larvae of the Tobacco Hornworm, Manduca sexta. AB - We examined growth rates, gas exchange patterns and energy metabolism of tobacco hornworm (Manduca sexta) larvae parasitized by the braconid wasp Cotesia congragata. Larvae parasitized at the beginning of the fourth-instar had reduced growth compared to unparasitized larvae of the same age and short-term differences in metabolism (measured as rates of CO(2) production, Vdot; CO(2)) were apparent almost immediately after wasp oviposition. However, over the growth period between parasitization and the last part of the fifth-instar, there was no significant difference between parasitized and unparasitized hosts as seen in the relationship between mass and Vdot; CO(2). One day prior to parasitoid emergence, host larvae stopped eating, ceased spontaneous locomotor activity and showed a dramatic decline in metabolism. The 60% decline of Vdot; CO(2) at this time is consistent with lack of specific dynamic action because the animals were not feeding. Gas exchange became highly cyclical on the day of parasitoid emergence, but the cause and significance of this phenomenon, which disappeared by the third day following emergence, are not clear. This pattern of cycling was not induced by starving nonparasitized larvae for 6days, nor by immobilizing nonparasitized larvae with tetrodotoxin. Ecdysteroid levels in the host's hemolymph significantly increased on the day when parasitoids completed their L2-L3 molt and began emerging, but not during the wasps' L1-L2 molt which occurred a few days earlier. Contrary to our initial expectation that hemolymph ecdysteroid titers might be linked to alterations in the host's metabolic rate, we observed no such correlation. PMID- 12769916 TI - Eicosanoids mediate microaggregation and nodulation responses to bacterial infections in black cutworms, Agrotis ipsilon, and true armyworms, Pseudaletia unipuncta. AB - Nodulation is the first, and quantitatively predominant, cellular defense reaction to bacterial infection in insects and other invertebrates. Inhibition of eicosanoid biosynthesis in true armyworms, Pseudaletia unipuncta, and black cutworms, Agrotis ipsilon, immediately prior to intrahemocoelic injections with heat-killed preparations of the bacterium, Serratia marcescens, severely impaired the nodulation response. Five eicosanoid biosynthesis inhibitors, including dexamethasone (a phospholipase A(2) inhibitor), indomethacin, ibuprofen (cyclooxygenase inhibitors), phenidone (dual lipoxygenase/cyclooxygenase inhibitor) and eicosatetraynoic acid (an arachidonic acid analog that inhibits all arachidonic acid metabolism) severely reduced nodulation in infected insects. The dexamethasone effects were reversed by treating true armyworms with arachidonic acid immediately after infection. In addition to these pharmacological findings, we demonstrate that an eicosanoid biosynthesis system is present in these insects. Arachidonic acid is present in fat body phospholipids at about 0.4% of total phospholipid fatty acids. Fat body expressed a phospholipase A(2) that can hydrolyze arachidonic acid from the sn-2 position of cellular phospholipids. Fat body preparations were competent to biosynthesize prostaglandins, of which PGE(2) was the major product. These findings support the hypothesis that eicosanoids mediate cellular immune reactions in insects. PMID- 12769919 TI - Glycogen in honeybee queens, workers and drones (Apis mellifera carnica Pollm.). AB - Honey bees (Apis mellifera carnica Pollm.) have low glycogen reserves in summer. Upon emergence drones have significantly larger amounts per unit weight when emerging, than workers; perhaps as adaption to the risk of not being fed as intensely as young workers. Maximum content was 0.23mg for workers (28d), and 0.59mg for drones (after emergence). Workers have relatively constant glycogen contents during their life, and very young drones have more glycogen than older ones. Young queens are similar to workers. In workers and queens in summer the greatest amounts of glycogen are found in the thorax. When the bees start flying (6th-8th day of life), drones have the highest amounts in the head (probably to supply their eyes), and upon maturity, drones have the least glycogen in the abdomen.Workers in winter show different glycogen values depending on whether they are active bees from the core area (0.23mg) or inactive ones from the outer surface of the winter cluster (0.37mg). They use glycogen from the thorax and the abdomen for their ongoing energy need. PMID- 12769921 TI - Defining Interaction between Electroantennogram Responses of Epiphyas postvittana (lepidoptera: Tortricidae) to Pheromone and Other Volatiles. AB - The potential for electroantennograms (EAGs) to assist in the measurement of atmospheric pheromone concentrations was examined in laboratory and field experiments by using multiple stimuli, the main component of the pheromone of Epiphyas postvittana, (E)-11-tetradecenyl acetate, its Z-isomer (a behavioral antagonist), and alpha-terpineol (a representative host-plant odor) were presented to the antenna simultaneously to simulate field conditions. The EAG results were compared with predictions from two models describing responses to combined stimuli. Responses are defined as log-additive if they can be described with the equation [EAG((total)) =a (log ([P(x)]+[B(y)])+c] and as linear-additive if the EAG follows the equation [EAG((total)) =a (log [P(x)])+c+a'(log [B(y)])+c'] where [B(y)] is the concentration of the stimulant background odor and [P(x)] is the concentration of an additional odor stimulus. The EAGs elicited by the added stimuli were inversely related to the concentration of the volatile in the background. EAGs elicited by all combined stimuli followed the log additive model. Our laboratory results were validated in field tests; alpha terpineol represents the volatiles present in orchard air. In spite of this interaction between the perception of pheromone and plant volatiles, the field EAG can be used for the measurement of atmospheric pheromone concentrations, where background odor concentrations are relatively constant. PMID- 12769920 TI - Two excitatory motoneurons differ in quantal content of their junctional potentials in abdominal muscle fibers of the cricket, Gryllus bimaculatus. AB - In abdominal muscles 202 and 203 of the cricket, Gryllus bimaculatus, large and small excitatory junctional potentials (l- and s-EJPs) with similar durations can be recorded from the same muscle fibers. At the normal extracellular calcium ion concentration ([Ca(2+)](o)) of 5mM, the amplitudes of l-EJPs in both muscles were larger than the threshold membrane potential for muscle action potentials, which is about -40mV. Below 0.75mM [Ca(2+)](o), the amplitudes became much smaller and were below the firing level for the action potentials. At 0.5mM, they fluctuated and decreased to 10.3 and 1.9mV in muscles 202 and 203, respectively, and at 0.25mM frequent failures occurred. The amplitudes of s-EJPs at 5mM [Ca(2+)](o) were 13.3 and 5.1mV in muscles 202 and 203, respectively, and the fluctuating amplitudes were far below the threshold for muscle action potentials. Below 0.75mM, s-EJPs were rarely observed. The relation between log(EJP amplitude) and log([Ca(2+)](o)) was linear within a certain range of [Ca(2+)](o) and the slopes of the lines for l-EJPs were about twice as steep as those for s-EJPs in both muscles. In muscle 202, the amplitude distribution of l-EJPs obtained at 0.25mM and that of s-EJPs at 0.75mM both showed peaks at once and twice the voltage at the first peak, which were coincident with the voltages at the peaks of amplitude distributions of miniature EJPs recorded simultaneously. The reversal potentials for l- and s-EJPs in muscle 202 were +1.02 and +0.22mV, respectively. In muscle 202, the decreases in amplitude of both EJPs by L-glutamate were similar and concentration-dependent. The results suggest that the difference in amplitude between l- and s-EJPs is attributable mainly to the difference in quantal contents. PMID- 12769922 TI - The effect of ooplasmic pH regulation on the formation of yolk spheres in the telotrophic ovariole of Dysdercus intermedius. AB - Vitellogenic oocytes of Dysdercus intermedius (Heteroptera: Pyrrhocoridae) were treated with the proton ionophore monensin in order to load the ooplasm with protons along the electrochemical gradient. Additionally, changes in the ooplasmic pH (DeltapH(OOC)) were recorded during exposure the oocytes to potassium-free medium (K(+)(MED)=0mM; choline for K(+)) or sodium-free medium (Na(+)(MED)=0mM; 40mM of choline for 40mM of Na(+)). The following observations were made: 1) The average ooplasmic pH (pH(OOC)) recorded during immersion in physiological saline solution (PSS) was pH(OOC(PSS))=7.40. 2) K(+)(MED) had no effect on pH(OOC) (pH(OOC(K-FREE)) congruent with pH(OOC(PSS))). 3) In sodium free medium the pH(OOC) decreased by H(+) influx in the magnitude of DeltapH(OOC(Na-FREE))=pH(OOC(Na-FREE))-pH(OOC(PSS))=-0.05 pH units. 4) The decreased pH(OOC) observed in sodium-free medium returned to initial values (7.40) by pumping out H(+) when 40mM of choline were replaced by 40mM of Na(+). 5) Addition of monensin (10&mgr;M; under the condition of Na(+)(MED)=0mM) reduced pH(OOC) in the magnitude of DeltapH(OOC(MON))=pH(OOC(MON))-pH(OOC(PSS))=-0.14. 6) Monensin induced ooplasmic proton loading was reversible when 40mM choline were replaced by 40mM Na(+).VITELLOGENESIS WAS DEMONSTRATED BY THE ACCUMULATION OF FLUORESCENCE LABELLED HEMOLYMPH PROTEINS IN YOLK SPHERES IN THE CORTEX OF THE OOCYTE: 1) Yolk formation continued in potassium-free medium. 2) The formation of yolk spheres came to a halt in sodium-free medium and, additionally, in the presence of monensin (10&mgr;M; Na(+)(MED)=0mM). 3) Breaks in yolk formation under the condition of Na(+)(MED)=0mM or during monensin treatment were stopped by replacing 40mM of choline with 40mM of Na(+). The results obtained using proton-specific microelectrodes and the in vitro assay to detect the formation of yolk spheres indicate that both the ooplasmic pH regulation and the acidification of vesicles during vitellogenesis are under control of a H(+)/Na(+) antiporter. PMID- 12769923 TI - Separation of Bombyxin from a neuropeptide of Bombyx mori showing Summer-morph producing Hormone (SMPH) activity in the Asian Comma Butterfly, Polygonia c aureum L. AB - A neuropeptide from brain-suboesophageal ganglion (Br-SG) complexes of the silkmoth, Bombyx mori, shows summer-morph-producing hormone (SMPH) activity in the Asian comma butterfly, P. c-aureum. The SMPH-active peptide was extracted and demonstrated to be almost the same molecular size as bombyxin (4-5kD), a nueropeptide which shows prothoracicotropic hormone (PTTH) activity when assayed in vitro with prothoracic glands (PGs) of 4th-instar B. mori larvae in vitro. A Sephadex G-50 fraction of 3-8kD molecules prepared from Br-SG complexes of B. mori adults was applied to CM-, SP-, DEAE- or QAE- Toyoperal columns at pH 5.6 (or pH 6.9). The SMPH-activity could be separated from the PTTH-activity (or bombyxin) by subjecting a SMPH- and PTTH-active preparation of B. mori to anion exchange chromatography at pH 6.9. By reversed-phase HPLC following an anion exchange chromatography, SMPH-activity was recovered in two fractions of 40-45% acetonitril. Results demonstrate that the B. mori peptide showing the SMPH activity in P. c-aureum is a different molecule than bombyxin. PMID- 12769924 TI - The Effects of Eicosanoid Biosynthesis Inhibitors On Prophenoloxidase Activation, Phagocytosis and Cell Spreading in Galleria mellonella. AB - The invertebrate immune system produces melanotic nodules in response to bacterial infections and this has previously been shown to be mediated by eicosanoids. Nodulation occurs in two phases: the first involves hemocyte degranulation and activation of the prophenoloxidase cascade; the second involves formation of a cellular capsule by attachment and spreading of hemocytes. We demonstrate that inhibitors of eicosanoid biosynthesis affect both of these phases of nodulation in Galleria mellonella. The phospholipase A(2) inhibitor, dexamethasone, as well as the cyclooxygenase inhibitor, indomethacin, significantly inhibit phagocytosis in vitro and prophenoloxidase activation in vivo. The inhibitory effects of dexamethasone were abolished by the addition of exogenous arachidonic acid. Furthermore, 5,8,11,14- eicosatetraynoic acid, dexamethasone and indomethacin inhibit hemocyte spreading in vitro. The findings support the idea that eicosanoid derivatives mediate both phases of the nodulation response and are consistent with previous studies which attribute roles for eicosanoids in other species as modulators of cell activity. PMID- 12769925 TI - Comparison of the autogenous potentials of newly-emerged anautogenous female mosquitoes. AB - The autogenous potentials in several species of newly emerged anautogenous female mosquitoes were examined by homoplastic transplantion of resting stage ovaries into newly-emerged females. Ovaries from 4-day-old female Culex pipiens quinquefasciatus, with follicles at stage Ib, were all activated in the newly emerged hosts and deposited some yolk in the oocyte. In addition, some of the implants (5-10%) matured. Transplanted C. tritaeniorhynchus ovaries also started to develop beyond the Ib resting stage, but never grew beyond the IIb stage. Resting Armigeres subalbatus and Aedes albopictus ovaries, both follicles having already reached the IIa-IIb stages, did not show any further growth after being implanted into newly emerged hosts. However, resting Ae. albopictus ovaries matured when heteroplastically transplanted into Culex pipiens molestus (74%) or into Culex pipiens pallens (5%). In addition, Ae. albopictus females, acting as hosts, could mature transplanted C. p. molestus (23%) and C. p. pallens ovaries (10%). Thus, the above specific differences in autogenous potentials seem to reside not only in the internal environment of the hosts, but also in the ovaries themselves. Development of the host ovaries in the above homoplastic and heteroplastic transplantations were usually suppressed by the implanted donor ovaries and failed to reach the resting stages of each species. PMID- 12769926 TI - Carbonic anhydrase supports electrolyte transport in Drosophila Malpighian tubules. Evidence by X-ray microanalysis of cryosections. AB - Electron probe X-ray microanalytical studies on the role of carbonic anhydrase in electrolyte transport in the cells of Drosophila Malpighian tubules indicate that carbonic anhydrase delivers protons and bicarbonate ions to ion transport systems in the cell membrane. After injection and after feeding acetazolamide or hydrochlorothiazide, known inhibitors of carbonic anhydrase, the contents of potassium, magnesium and chloride in the apical cytoplasm and in the cytoplasm close to the basal plasma membrane decreased. We explain our measurements by the hypothesis of a basal Mg-H-antiport system in parallel with Cl-HCO(3)-antiport, inhibitable by DIDS. Zinc is supposed to enters cells and intracellular Zn storage vacuoles by a negatively charged Zn-anion-complex in exchange for HCO(3)( ) ions. This antiport is inhibitable by SITS. The content of the Zn storage vacuoles is acid, as shown by red fluorescence after incubation of Malpighian tubules with acridine orange. Red fluorescence is absent after preincubation in a medium containing an inhibitor of carbonic anhydrase. Carbonic anhydrase was demonstrated cytochemically in the Golgi-ER complex, Golgi vesicles and intercellular space. We suppose that carbonic anhydrase is synthesized and stored in the Golgi-ER-complex from where it is released into the tubule lumen. PMID- 12769927 TI - Inhibition of testicular growth and development in Manduca sexta larvae parasitized by the braconid wasp Cotesia congregata. AB - Tobacco hornworm larvae parasitized by the gregarious larval endoparasitoid Cotesia congregata exhibited an inhibition in testicular growth and development, the extent of which was determined by the age and developmental stage of the host at the time of parasitization. The degree of parasitic castration, as assessed by measurements of testicular volume, was correlated with the stadium in which parasitization occurred. A mathematical formula requiring the measurement of testicular length, width and depth was used to calculate testicular volume. The use of the depth parameter revealed a negative correlation between host weight and testicular volume in parasitized larvae. Testicular volumes of fifth instar hosts, which had been parasitized in the first stadium, were significantly smaller than those originally parasitized as fourth or fifth instar larvae and were not correlated with parasitoid load. Effects of natural parasitism were not duplicated by injections of C. congregata polydnavirus and venom, topical treatment with the juvenile hormone analog methoprene, or starvation of nonparasitized larvae. Larvae receiving virus plus venom or methoprene grew larger due to delayed wandering and had larger testes than controls. Deleterious effects on host testes may be due to the effects of nutrient competition between the developing parasitoid progeny and the gonads, combined with the juvenilizing effects believed to be caused by the polydnavirus. PMID- 12769928 TI - Ultrastructural effects of lethal freezing on brain, muscle and Malpighian tubules from freeze-tolerant larvae of the gall fly, Eurosta solidaginis. AB - In preparation for winter low temperatures, larvae of the gall fly, Eurosta solidaginis, accumulate the cryoprotectants glycerol, sorbitol, and trehalose. The fat body cells of these freeze-tolerant larvae can survive intracellular freezing to -80 degrees C for 48 h even though no whole larvae survive this treatment. We hypothesized that some other tissue was more susceptible to freezing and therefore may be responsible for larval death. This paper compares the ultrastructure of brain, muscle, and Malpighian tubules between non-lethally frozen and lethally frozen freeze-tolerant larvae. The nuclei of cortical brain cells from lethally frozen larvae exhibited clumped chromatin and nuclear membranes with occasional expansions or 'blebs' of the intermembranous space, while the cytoplasm contained swollen spheres of endoplasmic reticulum. In contrast, non-lethally frozen brain contained nuclei with evenly dispersed chromatin, smooth nuclear membranes and a cytoplasm free of swollen endoplasmic reticulum. Muscle tissue of lethally frozen larvae contained disrupted myofilaments surrounding the Z-line in comparison to non-lethally frozen muscle which had myofilaments extending all the way to the Z-line. Alterations of Malpighian tubule cells from lethally frozen larvae included an extracted cytoplasm with swollen and rounded mitochondria. In contrast, Malpighian tubule cells from non-lethally frozen larvae had a more concentrated cytoplasm with many rod-shaped mitochondria. Results show alterations to all three tissue types due to lethal freezing. The brain tissue contained the most observable alterations and therefore may be the most susceptible to lethal freeze damage. PMID- 12769929 TI - Life cycle expression of a bombyxin-like neuropeptide in the tobacco hornworm, Manduca sexta. AB - Immunocytochemistry was used to investigate the developmental expression of the insulin-like neuropeptide bombyxin in the tobacco hornworm, Manduca sexta. A mouse monoclonal antibody raised against a synthetic peptide corresponding to bombyxin's A-chain N-terminus was used to localize a bombyxin-like peptide to a group of cerebral medial neurosecretory cells, the M-NSC IIa(2). Immunostaining was first detected on day 0 of the second larval instar, localized in the M-NSC IIa(2) somata and in the neurohemal organ, the corpora allata (CA). By day 0 of the fourth larval instar, the peptide was present throughout the M-NSC IIa(2) somata, axons, dendritic fields and CA. Between days 7 and 9 of the fifth instar, a dramatic reduction in the dendritic fields and CA staining occurred, suggesting the peptide is released. After day 2 of the pupal period, only M-NSC IIa(2) somata immunostained, a pattern that persisted through day 2 of the adult stage. The specificity of immunostaining was demonstrated by using a synthetic bombyxin peptide to block staining. These developmental data reveal times of potential Manduca bombyxin-like peptide release which should provide insight into the peptide's function. PMID- 12769930 TI - Ultrastructural Effects of a Non-Steroidal Ecdysone Agonist, RH-5992, on the Sixth Instar Larva of the Spruce Budworm, Choristoneura fumiferana. AB - Force feeding of RH-5992 (Tebufenozide), a non-steroidal ecdysone agonist to newly moulted sixth instar larvae of the spruce budworm, Choristoneura fumiferana, (Lepidoptera: Tortricidae) initiates a precocious, incomplete moult. Within 6h post treatment (pt) the larva stops feeding and remains quiescent. Around 12hpt, the head capsule slips partially revealing an untanned new head capsule that appears wrinkled and poorly formed. By 24hrpt, the head capsule slippage is pronounced and there is a mid-dorsal split of the old cuticle in the thoracic region but there is no ecdysis. The larva remains moribund in this state and ultimately dies of starvation and desiccation. The temporal sequence of the external and internal changes of the integument were studied using both scanning and transmission electron microscopy. Within 3hpt, there is hypertrophy of the Golgi complex indicating synthetic activity and soon after, large, putative ecdysial droplets are seen. Within 24h, a new cuticle that lacks the endocuticular lamellae is formed. The formation of the various cuticular components, the degradation of the old cuticle and changes in the organelles of the epidermal cells of the mesothoracic tergite are described. The difference between the natural moult and the one induced by RH-5992 are explained on the basis of molecular events that take place during the moulting cycle. The persistence of this ecdysone agonist in the tissues permits the expression of all the genes that are up-regulated by the presence of the natural hormone but those that are turned on in the absence of the hormone are not expressed. PMID- 12769931 TI - Physiological and Biochemical Analysis of Factors in the Female Venom Gland and Larval Salivary Secretions of the Ectoparasitoid Wasp Eupelmus orientalis. AB - The stinging adult female and the biting newly-hatched larva of the solitary ectoparasitoid wasp Eupelmus orientalis can both cause permanent paralysis and stop the development of Callosobruchus maculatus host larvae. These two processes of host envenomation appeared to be independent and complementary in primary parasitism or in hyperparasitism of a distantly related hymenopteran host species. In contrast, the development of larvae as hyperparasites on members of their own species or genus depended completely on the prior injection of female venom. The venoms of the female and the first instar larva had similar effects on the cellular metabolism of the primary hosts. Protein synthesis was blocked in C. maculatus hosts envenomated by a female or a first instar larva of E. orientalis, but the absence of DNA breakdown indicated that these paralysed hosts were alive and quiescent. The venomous secretions injected by adult females and first instar larvae of E. orientalis had distinct electrophoretic profiles. The immunoreactive features of proteins from female venom and larval secretions were also examined. There is evidence for antigenic conservation between some venom proteins of E. orientalis and Apis mellifera. Lastly, the hyaluronidase, phospholipase and lipase activities in the female venom gland and in larval-derived secretions of E. orientalis were assayed. No lipase activity was detected. Phospholipase activity was found in both the female venom and the larval secretions of E. orientalis, whereas hyaluronidase was specific to the female venom. PMID- 12769932 TI - Identification of Components of the Oviposition Aggregation Pheromone of the Gregarious Desert Locust, Schistocerca gregaria (Forskal). AB - Acetophenone and veratrole have been identified as two major behaviourally active components of the oviposition aggregation pheromone of the desert locust, Schistocerca gregaria. These compounds were identified from the volatiles of egg pod froth using gas chromatography-electroantennographic (GC-EAD) and gas chromatography-mass spectrometric (GC-MS) analysis. Both compounds were shown to elicit aggregation of gravid females in oviposition bioassays; however, they do not act synergistically. Both acetophenone and veratrole individually at optimum doses induced about 70% egg laying, a value similar to that evoked with froth volatiles. PMID- 12769933 TI - Digestion of Pollen Components by Larvae of the Flower-Specialist Bee Chelostoma florisomne (Hymenoptera: Megachilidae). AB - Pollen digestion in larvae of the solitary bee Chelostoma florisomne (Megachilidae), a pollen-specialist (oligolectic) species on Ranunculus spp. (Ranunculaceae), was investigated using histological techniques. Serial sections from the anterior-, median-, and posterior-midgut/hindgut regions of the larval alimentary canal were selectively stained and examined for disappearance of lipids, proteins, and carbohydrates from ingested Ranunculus pollen grains. During digestion, pollen contents were observed to gradually extrude through the grain apertures. Most of the digestion of lipids and proteins occurred in the anterior- and median-midgut, whereas carbohydrates disappeared more slowly and mainly in the median- and posterior-midgut. In the hindgut, pollen comprised mainly empty exine shells, which were crushed and often broken, suggesting that some components of the wall are degraded during digestion, causing collapse of the outer, chemically resistant wall layer (exine). Lipids were completely digested, but small quantities of proteins and carbohydrates were often still present in grains in the posterior midgut/hindgut. The pollenkitt appeared to be completely absorbed, with none evident in the posterior midgut or in larval feces. The findings indicate that this oligolectic bee has the digestive equipment necessary to effectively remove and absorb the nutrients present in the pollen of its restricted food source. Comparison with similar studies in adult honey bees suggests that differing digestive abilities might be an underlying factor in pollen specialization in bees. PMID- 12769934 TI - Effect of the Host Plant on the Antioxidative Defence in the Midgut of Lymantria dispar L. Caterpillars of Different Population Origins. AB - THE RESPONSES OF GYPSY MOTH LARVAE ORIGINATING FROM TWO POPULATIONS (OAK FOREST, LOCUST FOREST) TO FAVORABLE (OAK) AND UNFAVORABLE (LOCUST) HOST PLANTS WERE MONITORED AT THE LEVEL OF MIDGUT ANTIOXIDATIVE DEFENCE: the activities of superoxide dismutase (SOD), catalase (CAT), glutathione-S-transferase (GST), glutathione reductase (GR), glutathione peroxidase like ('GSH-Px like') and glutathione content (GSH). Short-term change of the diet (3 days) to locust leaves of the 5th instar larvae (oak population) provoked an increase in GST and 'GSH-Px like' activities as well as in the amount of GSH. On the contrary, transferring the gypsy moth larvae (locust population) to oak leaves was followed by a decrease in GST, 'GSH-Px like' activities, and in the amount of GSH. Feeding gypsy moth larvae from hatching on an unfavorable host plant such as locust, led to increases in GST and SOD activities and GSH content, as well as to a decrease in CAT activity in all instars studied (4th, 5th, 6th). The locust leaf diet caused changes in other components of antioxidative defence dependent on larval instar and population origin, a feature which could be ascribed to trophic adaptation of the gypsy moth to an unfavorable host plant. PMID- 12769935 TI - Immunohistochemical and developmental studies to elucidate the mechanism of action of the snowdrop lectin on the rice brown planthopper, Nilaparvata lugens (Stal). AB - Rice brown planthoppers (Nilaparvata lugens) were fed on artificial diet containing snowdrop lectin (Galanthus nivalis agglutinin; GNA), which has been shown to be toxic towards this insect pest. In addition to decreasing survival, the lectin affected development, reducing the growth rate of nymphs by approximately 50% when present at a concentration of 5.3&mgr;M. Immunolocalisation studies showed that lectin binding was concentrated on the luminal surface of the midgut epithelial cells within the planthopper, suggesting that GNA binds to cell surface carbohydrate moieties in the gut. Immunolabelling at a lower level was also observed in the fat bodies, the ovarioles, and throughout the haemolymph. These observations suggest that GNA is able to cross the midgut epithelial barrier, and pass into the insect's circulatory system, resulting in a systemic toxic effect. Electron microscope studies showed morphological changes in the midgut region of planthoppers fed on a toxic dose of GNA, with disruption of the microvilli brush border region. No significant proteolytic degradation of GNA was observed either in the gut or honeydew of planthoppers fed on lectin-containing diet. The presence of glycoproteins which bind GNA in the gut of the brown planthopper was confirmed using digoxigen labeled lectins to probe blots of extracted gut polypeptides. PMID- 12769936 TI - Geographical and diapause-related cold tolerance in the blow fly, Calliphora vicina. AB - Three geographical strains of the blow fly, Calliphora vicina, were tested for cold tolerance at 0 degrees, -4 degrees and -8 degrees C. Survival to eclosion after 1 to 18 days of cold exposure was greater for diapause-destined larvae than for nondiapause-destined larvae of the two northern strains (Nallikari, Finland 65 degrees N and Edinburgh, Scotland 55 degrees N) but not for the southernmost strain (Barga, Italy 44 degrees N) where no clear differences were apparent. Diapause-destined larvae of the Edinburgh strain were more cold tolerant than those from Nallikari, at both -4 degrees and -8 degrees C, a difference possibly attributable to the long-lasting snow cover in the more northern locality, which might insulate the overwintering soil microclimate. At 0 degrees C, however, Nallikari larvae were more cold tolerant than Edinburgh or Barga. This was also the case for nondiapause-destined larvae, indicating that cold tolerance may occur, in part, independently of the diapause programme. In all three strains diapausing larvae were more cold tolerant than same-age (nondiapausing) pupae. For Nallikari, but not Barga, wandering larvae from short-day exposed flies, therefore initially programmed for diapause, but diverted from the diapause pathway by larval breeding at 19 degrees C, were significantly more cold tolerant than nondiapause larvae from long-day parents, indicating some maternal regulation of larval cold tolerance. There was, however, no evidence for an additional cold hardiness in larvae acclimatised to cold by a gradual reduction of temperature. PMID- 12769937 TI - Role of the head in the ultrastructural midgut organization in Rhodnius prolixus larvae: evidence from head transplantation experiments and ecdysone therapy. AB - Studies on the effects of decapitation, head transplantation and ecdysone therapy on the ultrastructural organization of the midgut in 5th-instar larvae of Rhodnius prolixus, were carried out. Control insects had a typical and significant organization of the epithelial cells (mainly microvilli, extracellular membrane layers and basal portion of the epithelial cells) of the midgut (stomach and intestine) during the entire period of the experiment. However, the host larvae, when decapitated 1 day after feeding, demonstrated significant changes in the ultrastructural organization of the epithelial cells of these compartments. In converse experiments, head transplantations from untreated donors 4-5 days after feeding into headless larvae sustained the ultrastructural organization of the epithelial cells in the midgut. Oral therapy with ecdysone (5 &mgr;g/mL of blood meal) in decapitated insects significantly reversed the altered organization of the stomach and intestine. These results point to a brain factor, possibly the prothoracicotropic hormone (PTTH) which stimulates ecdysteroid production in the prothoracic glands, may be a factor responsible, directly or indirectly, for the midgut cell organization in R. prolixus. PMID- 12769938 TI - Anti-diuresis in the blood-feeding insect Rhodnius prolixus Stal: antagonistic actions of cAMP and cGMP and the role of organic acid transport. AB - Secretion of primary urine by upper Malpighian tubules of the blood-sucking insect Rhodnius prolixus has recently been shown to be inhibited by cyclic GMP (cGMP). In the present work, we have demonstrated that cGMP has effects antagonistic to those of cAMP in Rhodnius tubules and have further characterized the effects of cGMP on tubular secretion. Cyclic GMP inhibited secretion at all concentrations from 5x10(-6) to 10(-3)M, though this inhibition was partially or wholly reversed by large (2mM) doses of cAMP. While sub-maximal concentrations of cGMP did not significantly alter [K(+)] and [Na(+)] of secreted fluid, high external [cGMP] reduced secretion to minimal levels and caused [K(+)] and [Na(+)] to approach pre-stimulation levels. Cyclic GMP does not appear to affect the permeability of the lower Malpighian tubule to water. Both cAMP and cGMP likely enter tubule cells by way of an organic acid transporter whose activity is induced by feeding. Sensitivity of the tubules to exogenous cGMP and cAMP, which is assumed to be a function of transport activity, reaches a peak approximately 5 days after the blood meal and declines rapidly thereafter. Transport of anions into upper tubules involves at least two different transporters: one for acylamides (e.g., p-aminohippuric acid) and another for sulphonates (e.g., amaranth, phenol red). Amaranth and phenol red blocked the actions of both cGMP and cAMP, whereas p-aminohippuric acid was without effect. This suggests that cyclic nucleotides enter by way of the sulphonate transporter. PMID- 12769939 TI - Two strains of cabbage seed weevil (Coleoptera: Curculionidae) exhibit differential susceptibility to a transgenic oilseed rape expressing oryzacystatin I. AB - The aim of this study was to assess the potential effect of a transgenic line of oilseed rape expressing oryzacystatin I (OCI) on two strains of cabbage seed weevil. The level of OCI expression in seeds was approximately 0.05% of total soluble proteins. The insects were field-collected in two different locations, and their progeny was analyzed after a 3 week-development in pods. Both strains showed a similar pattern of proteolytic activity, and similar levels of OCI sensitive proteinase activity in vitro. However, the larvae showed differential susceptibility to the transgenic plants. Despite inhibition of digestive proteinases in vitro by OCI in both strains, one strain showed an increased growth rate when fed the transgenic seeds, while the other strain remained unaffected. While suggesting the importance of studying individuals from different populations when assessing the effect of proteinase inhibitor expressing plants on insect growth, our results also point out the necessity of studying the biochemical interactions taking place in vivo between the recombinant inhibitors and their target proteinases. PMID- 12769940 TI - Nectar feeding by the ant Camponotus mus: intake rate and crop filling as a function of sucrose concentration. AB - In independent assays, workers of the ant Camponotus mus were conditioned to visit an arena where they found a large drop of sucrose solution of different concentrations, from 5 to 70% weight on weight (w/w). Single ants were allowed to collect the sucrose solution ad libitum, and feeding time, feeding interruptions, crop load, and intake rates were recorded. Feeding time increased exponentially with sucrose concentration, and this relationship was quantitatively described by the increase in viscosity with concentration corresponding to pure sucrose solutions. Ants collecting dilute solutions (5 to 15% w/w) returned to the nest with partial crop loads. Crop filling increased with increasing sucrose concentration, and reached a maximum at 42.6% w/w. Workers collecting highly concentrated solutions (70% w/w) also returned to the nest with a partially filled crop, as observed for dilute solutions. Nectar intake rate was observed to increase with increasing sucrose concentration in the range 5 to 30% sucrose. It reached a maximum at 30.8%, and declined with increasing sucrose concentration. Results suggest that both sucrose concentration and viscosity of the ingested solution modulate feeding mechanics as well as the worker's decision about the load size to be collected before leaving the source. PMID- 12769941 TI - Light and temperature cooperate to regulate the circadian locomotor rhythm of wild type and period mutants of Drosophila melanogaster. AB - In the wild type (Canton-S) and period mutant flies of Drosophila melanogaster, we examined the effects of light and temperature on the circadian locomotor rhythm. Under light dark cycles, the wild type and per(S) flies were diurnal at 25 degrees C. However, at 30 degrees C, the daytime activity commonly decreased to form a rather nocturnal pattern, and ultradian rhythms of a 2 approximately 4h period were observed more frequently than at 25 degrees C. The change in activity pattern was more clearly observed in per(0) flies, suggesting that these temperature dependent changes in activity pattern are mainly attributable to the system other than the circadian clock. In a 12h 30 degrees C:12h 25 degrees C temperature cycle (HTLT12:12), per(0) flies were active during the thermophase in constant darkness (DD) but during the cryophase in constant light (LL). The results of experiments with per(0);eya flies suggest that the compound eye is the main source of the photic information for this reversal. Wild type and per(0) flies were synchronized to HTLT12:12 both under LL and DD, while per(S) and per(L) flies were synchronized only in LL. This suggests that the circadian clock is entrainable to the temperature cycle, but the entrainability is reduced in the per(S) and per(L) flies to this particular thermoperiod length, and that temperature cycle forces the clock to move in LL, where the rhythm is believed to be stopped at constant temperature. PMID- 12769942 TI - A thermoprotective role for sorbitol in the silverleaf whitefly, Bemisia argentifolii. AB - Accumulation of polyols in insects is well known as a cold-hardening response related to overwintering or to protection against cold shock. The silverleaf whitefly (Bemisia argentifolii, Bellows and Perring) is a major insect pest in tropical and subtropical regions where heat stress and desiccation pose formidable threats to survival. We found that sorbitol levels increased ten-fold when whiteflies were exposed to elevated temperatures. Sorbitol levels rose from 0.16nmolwhitefly(-1) at 25 degrees C to 1.59nmolwhitefly(-1) at 42 degrees C. Sorbitol levels fluctuated diurnally under glasshouse and field conditions increasing ten-fold from morning to early afternoon. Feeding experiments on artificial diets showed that both temperature and dietary sucrose concentration were key factors influencing sorbitol accumulation. Cell free extracts prepared from adult whiteflies catalyzed NADPH-dependent fructose reduction, but were unable to reduce glucose with either NADPH or NADH. Radiotracer experiments with labeled glucose and fructose showed that fructose was the immediate precursor of sorbitol. Thus, sorbitol synthesis in the whitefly is apparently unconventional, involving conversion of fructose by a novel NADPH-dependent ketose reductase. We propose that sorbitol accumulation is a mechanism for thermoprotection and osmoregulation in the silverleaf whitefly, allowing the insect to thrive in environments conducive to thermal and osmotic stress. PMID- 12769943 TI - Developmental changes in dopamine levels in larvae of the fly Chymomyza costata: comparison between wild-type and mutant-nondiapause strains. AB - Dopamine and two related catecholamines, L-3,4-dihydroxyphenylalanine (DOPA) and N-acetyl dopamine (NADA), were analyzed in whole body and tissue samples taken throughout late larval development of the drosophilid fly Chymomyza costata, which enters facultative diapause as a mature 3rd instar larva in response to short photophase. Wild-type (W) and mutant-nondiapause (M) strains reared under diapause inducing (d) and preventing (nd) photophases were compared. Developmental changes in the whole body of dopamine levels showed some general features irrespective of fly strain and rearing conditions: sharp major peaks during moult from second to third instar larva and during pupariation; lower minor peaks around the middle of both 2nd and 3rd instars. Significant differences between the strains and conditions were also found: dopamine levels were lower throughout the 2nd instar and during the 2nd to 3rd instar moult of mutant strain larvae (M/d) as compared to wild-type larvae (W/nd and W/d); while the late 2nd and late 3rd instar larvae destined to diapause (W/d) maintained relatively high dopamine concentrations, their counterparts destined to continuous development (W/nd and M/d) significantly decreased dopamine levels prior to the 2nd to 3rd instar moult or pupariation. Possible relationship between the dopamine levels and diapause induction/onset in C. costata larvae is discussed. Integument contained more than 90% of the dopamine found in the whole body. The gut and central nervous tissues showed relatively low pools of dopamine, only trace amounts were detected in haemolymph and no dopamine was found in fat body. DOPA levels were low and stable throughout larval development of both W and M strains and under both conditions. NADA levels peaked during second halves of 2nd and 3rd instars of both strains, then dropped to trace levels and were elevated again during 2nd to 3rd instar moult as well as in tanned prepupae. No elevation of NADA levels was recorded in 3rd instar W/d larvae which entered diapause. PMID- 12769944 TI - Interactions between desiccation resistance, host-plant contact and the thermal biology of a leaf-dwelling sub-antarctic caterpillar, Embryonopsis halticella (Lepidoptera: Yponomeutidae). AB - During May 1997 thermal tolerance, supercooling point (SCP), low and high temperature survival, and desiccation resistance were examined in field-fresh Embryonopsis halticella Eaton larvae from Marion Island. SCPs were also examined in acclimated larvae, larvae starved for seven days, larvae within their leaf mines, and in larvae exposed to ice crystals. Field-fresh larvae had a critical minimum temperature (CT(Min)) and critical maximum temperature (CT(Max)) of 0 degrees C and 39.7 degrees C, respectively. Mean SCP of field-fresh caterpillars was -20.5 degrees C and this did not change with starvation. Field-fresh larvae did not survive freezing and their lower lethal temperatures (70% mortality below -21 degrees C) and survival of exposure to constant low temperatures (100% mortality after 12hrs at -19 degrees C) indicated that they are moderately chill tolerant. SCP frequency distributions were unimodal for field-fresh larvae, but became bimodal at higher acclimation temperatures. Contact with ice-crystals caused an increase in SCP (-6.5 degrees C), but contact with the host plant had less of an effect at higher subzero temperatures. It appears that the remarkable desiccation resistance of the larvae is selected for by the absence of a boundary layer surrounding their host plant, caused by constant high winds. This suggests that the low SCPs of E. halticella larvae may have evolved as a consequence of pronounced desiccation resistance. PMID- 12769945 TI - Host cell allometry and regulation of the symbiosis between pea aphids, Acyrthosiphon pisum, and bacteria, Buchnera. AB - The symbiotic bacteria Buchnera in aphids are borne in cells, called bacteriocytes, in the insect haemocoel. The number and median volume of bacteriocytes in pre-reproductive adult insects varied significantly among 14 parthenogenetic clones of the pea aphid Acyrthosiphon pisum. After logarithmic transformation of the data, the relationship of both number and median volume of bacteriocytes with aphid weight for the clones could be described by common regression lines with slopes significantly greater than zero. The allometric slope for median bacteriocyte volume was calculated as 1.06, by model I regression and 1.94 by model II regression; and the equivalent values of the allometric slope for total volume of bacteriocytes were 1.51 and 2.50, suggesting that the total volume of bacteriocytes increases disproportionately with aphid body weight. The partial correlation coefficient between the number and median volume of bacteriocytes was +0.07, with body weight held constant. It is proposed that the regulation of number and size of bacteriocytes is not linked and that bacteriocytes may not exhibit compensatory changes in size, in response to alteration in number. Experimental manipulation of the rates of bacteriocyte differentiation and division could therefore perturb the total volume of the symbiosis, on which aphid pests depend for normal growth and reproduction. PMID- 12769946 TI - Ecdysteroids regulate yolk protein uptake by Drosophila melanogaster oocytes. AB - Juvenile hormones (JHs) are thought to drive the regulation of yolk protein uptake by ovaries in Drosophila melanogaster. However, the level of JH production in a mutant stock (ap(56f)) is depressed yet the flies are normally vitellogenic. The production of ecdysteroids by these ap(56f) ovaries in vitro is elevated above that of wild-type ovaries. The incubation of wild-type ovaries in the presence of 0.1mM JHB(3) increased ecdysteroid biosynthesis only during the first 18h following eclosion. Female Drosophila melanogaster undergo a pre-vitellogenic reproductive diapause when exposed to low temperature (11 degrees C) and a short day photoperiod (L12:D12). The rate of ecdysteroid synthesis by the ovaries, but not JH production, increased within 12h of a temperature upshift to 25 degrees C from a basal level of 20+/-1pg/10 pair of ovaries/5h to a sustained level of 150+/-20pg/10 pair/5h. Vitellogenic oocytes were noted in all females within 12h of this temperature upshift. Diapause was also terminated by the injection of 1&mgr;g of 20-hydroxyecdysone into the abdomens of diapausing females as determined by an increase in ovary size, and the appearance of vitellogenic oocytes as compared to controls. These results are consistent with a revised model for the regulation of yolk protein uptake by ovaries in which ecdysteroids, and not JHs, play the prominent role. PMID- 12769947 TI - Significance of methanogenic symbionts for development of the American cockroach, Periplaneta americana. AB - The effects of suppression of methanogenesis with a drug, 2-bromoethanesulfonic acid (BES), on the hindgut ecosystem and development of the cockroach Periplaneta americana fed either low or high fiber diet were evaluated by measuring methane production, volatile fatty acids (VFA) concentrations in the hindgut, cockroach weight gain and development time (the length of nymphal period). Methane production and VFA concentrations in the hindgut of cockroaches fed high fiber diet were significantly higher than those fed low fiber diet. Although BES treatment greatly reduced methane production, VFA concentrations in the hindgut, cockroach weight gain and development time were not significantly altered. These results indicate that methanogenic microbes are not essential for keeping low hydrogen pressure in the hindgut lumen, and normal cockroach development. PMID- 12769948 TI - Regulation of sex pheromone biosynthesis in the oriental tobacco budworm, Helicoverpa assulta (Lepidoptera: Noctuidae). AB - The five components, Z9-16:Ald, 16:Ald, Z11-16:Ald, Z9-16:Ac and Z11-16:Ac, of the sex pheromone in Helicoverpa assulta were mostly detected during the scotophase, with their titer peaking at the 4th hour during the scotophase under a 15L/9D regime. They were not detected during the photophase, but were produced during the photophase when decapitated females were injected with extracts of virgin female (FHE), male heads (MHE), homogenates of the brain-suboesophageal ganglion complex (Br-SOG), or synthetic Hez-PBAN. Production of Z9-16:Ald increased during the first 45min after FHE injection and then declined to a very low level after 2h during the photophase. Synthetic Hez-PBAN stimulated the sex pheromone glands for at least 2h and the effect was more or less proportional to the concentration of the peptide. From the present results, we suggest the following: PBAN is released continuously into the haemolymph to stimulate pheromone biosynthesis at least during the first half of the scotophase, PBAN is synthesized and accumulated independent of photoperiod or sex, and the release starts just prior (about 1h) to the beginning of the scotophase. PMID- 12769949 TI - Development of specific responses in antennal taste hairs after ecdysis. An electrophysiological investigation of the cockroach, Periplaneta brunnea. AB - Spike generation in cockroach antennal taste hairs of nymphs appears to follow the same rules as in adults analysed previously. After ecdysis, taste hairs on the proximal segments responded earlier than those on distal ones. Shorter hairs became active before longer ones. The mechanoreceptor responded before the chemoreceptors. During a transient period, 3-5 hours after ecdysis, all four chemoreceptors and also the mechanoreceptor were sensitive to 150mM KCl. Spikes elicited by 75 mM KCl were generated predominantly in the outer dendritic segments. They did not differ in shape and frequency when 60mM sucrose was added. The earliest sign of a specific reaction to sucrose was observed approximately 4 5 hours after ecdysis, when the mixture of sucrose and KCl elicited more spikes than KCl alone. PMID- 12769950 TI - Male orientation to trail sex pheromones in parasitoid wasps: does the spatial distribution of virgin females matter? AB - We studied male locomotory response to trails and patches of sex pheromone (left respectively by free-ranging females and females constrained to stay on a small area) in the two parasitoids Aphelinus asychis (Hymenoptera: Aphelinidae) and Trichogramma brassicae (Hymenoptera: Trichogrammatidae). Under the hypothesis that the spatial distribution of virgin females differs between these species (scattered among host plants in A. asychis, gregarious at emergence sites in T. brassicae), we predicted that male locomotory response to their sex pheromones should also differ: A. asychis males should follow pheromone trails on plants in order to encounter the females along these trails, whereas T. brassicae males should stay on pheromone patches, at emergence sites, and mate the females on these patches. Using an improved video-tracking system, we found that males of both species respond to conspecific sex pheromone trails and patches, but that the response does not differ much between species. Males released on marked substrates walked in a more convoluted pattern (i.e. higher path fractal dimension and higher number of crossings within tracks) than males released on unmarked substrates. On pheromone patches, males turned persistently in the same direction when leaving the patch, which explains a higher number of visits on marked patches than on unmarked patches, and possibly, higher track convolution on pheromone trails. Contrary to our hypothesis, male A. asychis did not follow female trails more accurately than male T. brassicae, and male T. brassicae did not stay longer on pheromone patches than male A. asychis. We argue that these discrepancies between our predictions and the observed responses originates from discrepancies between the assumed spatial distribution of virgin females and their actual distribution in the wild. PMID- 12769951 TI - Antennal morphology as a physical filter of olfaction: temporal tuning of the antennae of the honeybee, Apis mellifera. AB - There are many different antennal morphologies for insects, yet they all have the same functional role in olfaction. Chemical signals are dispersed through two physical forces; diffusion and fluid flow. The interaction between antennal morphology and fluid flow generates a region of changing flow velocity called the boundary layer. The boundary layer determines signal dispersion dynamics and therefore influences the signal structure and information that arrives at the receptor cells. To investigate how the boundary layer changes the information in the signals arriving at receptor cells, we measured chemical dynamics within the boundary layer around the bee antennae using microelectrodes. We used two types of chemical signals: pulsed and continuous. The results showed that the boundary layer increased the decay time of the chemical signal for the pulsatile stimuli and increased the peak height for the continuous data. Spectral analysis of continuous signals showed that the temporal aspects of the chemical signal are changed by the boundary layer. Particularly the temporal dynamics of the signal are dampened at the slowest flow speed and amplified at the intermediate and fast flow speeds. By altering the structure of the chemical signal, the morphology will function as a sensory filter. PMID- 12769952 TI - Queen mandibular gland pheromone influences worker honey bee (Apis mellifera L.) foraging ontogeny and juvenile hormone titers. AB - SYNTHETIC QUEEN MANDIBULAR GLAND PHEROMONE (QMP) WAS APPLIED TO HONEY BEE COLONIES TO TEST TWO HYPOTHESES: (i) QMP acts like a primer pheromone in the regulation of age-related division of labor, and (ii) this primer effect, if present, varies in three strains of workers that show genetically-based differences in their retinue attraction response to QMP (a pheromone releaser effect). Strains of workers that were high, or low in their response to QMP in a laboratory bioassay, as well as unselected 'wild-type' workers, were fostered in queenright colonies with or without supplemental QMP. Effects of QMP on foraging ontogeny and juvenile hormone III (JH) blood titers in worker honey bees were measured. Bees in QMP-supplemented colonies showed significant delays in foraging ontogeny, and foraging activity was reduced. They also had significantly lower JH titers, although the titer curves were somewhat atypical. There were no differences in foraging ontogeny or JH titers among the three strains. We conclude that (i) QMP can delay the ontogeny of foraging by some mechanism that suppresses JH production, (ii) this QMP primer response is independent of the retinue releaser response, and (iii) QMP can play an important role in regulating division of labour. PMID- 12769953 TI - Gut physicochemistry of grassland grasshoppers. AB - We examined the pH and Eh of the digestive tract of 23 species of mixed-grass prairie grasshoppers, and asked whether these traits were associated with the species breadth and forb composition of their diets. We report that the gut lumen of all grasshoppers was oxidizing and ranged from slightly acid to neutral depending on the gut region and species. Although gut physicochemical conditions differed among species, the differences were of small magnitude. Conditions were fairly uniform along the digestive tract, which suggests little or no regulation of pH or Eh. Gut conditions were independent of diet breadth and the percentage of forbs in the diet. These results suggest that physicochemical conditions of grasshopper guts are not highly regulated and are not influenced by their most recent meal or by broad scale patterns of host-plant use. PMID- 12769954 TI - Insect Na(+)/K(+)-ATPase. AB - Na(+)/K(+)-ATPase (sodium/potassium pump) is a P-type ion-motive ATPase found in the plasma membranes of animal cels. In vertebrates, the functions of this enzyme in nerves, heart and kidney are well characterized and characteristics a defined by different isoforms. In contrast, despite different tissue distributions, insects possess a single isoform of the alpha-subunit. A comparison of insect and vertebrate Na(+)/K(+)-ATPases reveals that although the mode of action and structure are very highly conserved, the specific roles of the enzyme in most tissues varies. However, the enzyme is essential for the function of nerve cells, and in this respect Na(+)/K(+)-ATPase appears to be fundamental in metazoan evolution. PMID- 12769955 TI - Partial desiccation induced by sub-zero temperatures as a component of the survival strategy of the Arctic collembolan Onychiurus arcticus (Tullberg). AB - The mechanism by which the freeze susceptible Arctic collembolan Onychiurus arcticus survives winter temperatures of -25 degrees C in the field is not fully understood but exposure to sub-zero temperatures (e.g. -2.5 degrees C) is known to induce dehydration and lower the supercooling point (SCP). In this study, changes in the water status and certain biochemical parameters (measured in individual Collembola) during a 3-week exposure to decreasing temperatures from 0 to -5.5 degrees C were studied. Osmotically active and inactive body water contents were measured by differential scanning calorimetry (DSC), water soluble carbohydrates by high performances liquid chromatography (HPLC) and glycogen by enzymatic assays. The activity of trehalase and trehalose 6-phosphate synthase were also measured. During the experiment, total water content decreased from 70 to 40% of fresh weight, mostly by the loss of osmotically active water with only a small reduction in the osmotically inactive component. The SCP decreased from 7 to -17 degrees C. Analysis of the results shows that if O. arcticus is exposed to -7 degrees C in the presence of ice, all osmotically active water would be lost due to the vapour pressure gradient between the animals supercooled body fluids and the ice. Under these conditions the estimated SCP would reach a minimum of c. -27 degrees C, but the Collembola may never freeze as all the osmotically active water has been lost, the animal becoming almost anhydrobiotic. Trehalose concentration increased from 0.9 to 94.7&mgr;g mg(-1)fw while glycogen reserves declined from 160 to 7.7 nmol glucose equivalents mg(-1) protein. Trehalase activity declined as the temperature was reduced, while trehalose 6 phosphate activity peaked at 0 degrees C. By adopting a strategy of near anhydrobiosis induced by sub-zero temperatures, O. arcticus, which was previously thought to be poorly adapted to survive severe winter temperatures, is able to colonise high Arctic habitats. PMID- 12769956 TI - Expression of actin in the central nervous system is switched off during diapause in the gypsy moth, Lymantria dispar. AB - Diapause-regulated proteins were identified in the CNS of pharate 1st instar larvae of the gypsy moth, Lymantria dispar. Expression of two proteins (130kDa and 45kDa) declined at the time of diapause initiation and remained low during diapause. At diapause termination, following exposure to 5 degrees C for at least 60 days, the 45-kDa protein was again highly expressed. Treatment of young pharate larvae with KK-42 averted diapause, and in such larvae, continued synthesis of the 45 kDa protein in the CNS was observed. These results suggest that expression of the 45-kDa protein is strongly down-regulated in the central nervous system (CNS) during diapause. Partial amino acid sequence determination suggested that the 45-kDa protein is actin, and this was confirmed using anti actin antibodies. RT-PCR using primers designed from the sequence of the gypsy moth midgut actin mRNA indicated that appearance of actin mRNA in the CNS followed the same pattern as that of the 45-kDa protein. The results indicate that diapause controls actin gene expression in the gypsy moth CNS and regulation is at the transcriptional level. PMID- 12769957 TI - Effects of nicotinic and muscarinic ligands on embryonic neurones of Periplaneta americana in primary culture: a whole cell clamp study. AB - The pharmacological properties of acetylcholine (ACh) receptors of cultured neurones from embryonic cockroach brains were studied using the whole-cell configuration of the patch-clamp technique. More than 90% of the studied neurones responded to ACh by a monophasic inward current, the intensity of which varied from cell to cell. The sequence of potency of the five tested agonists was ACh > nicotine=carbamylcholine > suberyldicholine=oxotremorine. The dose-response relationship was complex, suggesting the existence of two populations of receptors: high-affinity receptors (extrapolated K(d) around 10(-7) M) and low affinity receptors (extrapolated K(d) around 5x10(-5) M). The current-voltage relationship of the induced current was linear between -80 and -40 mV and the extrapolated reversal potential was not significantly different from 0 mV. The sequence of decreasing potency of the antagonists of the ACh response was: methyllycaconitine > alpha-bungarotoxin > mecamylamine > curare > strychnine > bicuculline > atropine > picrotoxin. These results show: (1) that, in embryonic brain neurones, the response to ACh corresponds to the opening of non-selective cationic channels; and (2) that the pharmacology of the ACh receptors is mainly but not solely nicotinic. The nature of the single events which underlie this response, as well as the structure of the channels (homo or hetero-oligomeric) remain to be investigated. PMID- 12769958 TI - Jaws that snap: control of mandible movements in the ant Mystrium. AB - Ants of the genus Mystrium employ a peculiar snap-jaw mechanism in which the closed mandibles cross over to deliver a stunning blow to an adversary within about 0.5 ms. The mandible snapping is preceded by antennation and antennal withdrawal. The strike is initiated by contact of the adversary with mechanosensory hairs at the side of the mandible, and is powered by large yet slow closer muscles whose energy is stored by a catapult mechanism. Recording of closer muscle activity indicates that the mandibles are not triggered by any fast muscle. Instead, we suppose that activity differences between the left and right mandible muscles imbalance a pivot at the mandible tip and release the strike. The likelihood for the strike to occur can be modulated by an alarm pheromone. The presence of specialized sensilla and of a complex muscle receptor organ shows that the mandibles are also adapted to functions other than snapping and suggests that the force of the mandible can be finely adjusted for other tasks. PMID- 12769959 TI - Expression of a male accessory gland peptide of Leptinotarsa decemlineata in insect cells infected with a recombinant baculovirus. AB - The male accessory glands (MAGs) of Leptinotarsa decemlineata produce an 8kDa peptide, designated Led-MAGP, that is recognized by monoclonal antibody MAC-18. The site of synthesis, amino acid sequence and the gene encoding this peptide have been documented ([Smid and Schooneveld, 1992][Smid et al., 1997]). The primary structure is homologous to the N-terminal hexa-repeat section of the chicken prion protein ([Harris et al., 1991]). The biological function of the Led MAGP has yet to be determined. For further research, large amounts of Led-MAGP is required, both for the production of a more specific antiserum, as well as for application in bio-assays. This paper describes the expression of Led-MAGP in insect cells infected with recombinant baculovirus, and the production of a polyclonal antibody against this recombinant peptide. The peptide was expressed under the control of the polyhedrin promotor. The resulting product was HPLC purified, and analysis on Western blots immuno-labelled with MAC-18 confirmed that the correct peptide was produced. Purified recombinant peptide was also analyzed by Edman degradation and mass spectrometry; this indicated that it was N terminally blocked and that the methionine residue at position 7 was oxidized. Large scale production resulted in the formation of aggregations of Led-MAGP, nevertheless a substantial proportion remained in a soluble state and could be harvested. A polyclonal antiserum encoded #87 was produced against recombinant Led-MAGP and its specificity was tested on Western blots of authentic peptide and on LM and EM sections of MAGs. All labelling results were equal to those obtained after MAC-18 labelling. However, antiserum #87 proved to be superior compared to MAC-18, since it recognizes the MAG peptide in normally fed, sexually active males, whereas MAC-18 labelling can only be accomplished after 7 days of starvation of the males. Therefore, the new antiserum #87 enables us to study the transfer dynamics of the Led-MAGP on histological sections. PMID- 12769960 TI - Growth stimulation of beetle larvae reared on a transgenic oilseed rape expressing a cysteine proteinase inhibitor. AB - The resistance of a transgenic line of oilseed rape expressing constitutively the cysteine proteinase inhibitor oryzacystatin I (OCI) was assessed against Psylliodes chrysocephala L. (Coleoptera: Chrysomelidae). The levels of OCI expression in the transformed line averaged 0.2% and 0.05% of total soluble protein in leaves and petioles respectively. In vitro analyses showed that P. chrysocephala larvae use both cysteine and serine proteinases for protein digestion, and that all the cysteine proteolytic activity is OCI-sensitive. However, bioassays showed that adults fed identically on leaf discs from control or transformed plants. When larvae were reared on transgenic plants expressing OCI, they showed an increase in weight gain compared to those reared on control plants. Furthermore, those larvae from transgenic plants exhibited a 2-fold increase in both cysteine and serine proteolytic activity as a reponse to the presence of OCI. The plasticity of insect digestive physiology and feeding behaviour are discussed, as well as the relevance of engineering a genotype expressing both types of proteinase inhibitors. PMID- 12769961 TI - Response to photoperiod during diapause development in the spider mite Tetranychus urticae. AB - To study the question whether photoperiodic time measurement in the spider mite Tetranychus urticae is based on a qualitative or quantitative principle, the duration of diapause development was determined in individual females at various constant photoperiods at 19 degrees C. Diapause duration at all four long-night treatments fluctuated around 64.5 days, varying from 62.2 at LD 12:12h to 66.4 at LD 10:14h. The within-treatment variation in diapause duration of the long-night groups appeared to be significantly correlated to the nightlength of the photoperiods used; the longer the nightlength, the higher the within-treatment variation. Frequency distributions of females completing diapause under the two regimes with nightlengths near the critical nightlength were skewed to the right. Mean diapause durations at these regimes, LD 13:11h and LD 14:10h, were 25.4 and 11.9 days, respectively. Mites completed diapause rapidly and synchronously under the three short-night photoperiods tested; within two weeks after transfer from cold storage at 4 degrees C to the diapause terminating regimes at 19 degrees C all females started reproduction. Mean diapause durations were 8.1, 6.4 and 6.5 days for the short-night treatments LD 15:9h, LD 17:7h and LD 19:5h, respectively. The coefficients of variation of diapause duration (variability within groups relative to the mean) of the short-night and the long-night groups varied from 18 to 42%; the coefficients of the two intermediate groups were 69and 81%. There was a clear difference in diapause duration between long-night and short-night groups, but no significant difference was present in this characteristic between different long-night groups on the one hand and only a small difference between different short-night groups on the other. These results support the hypothesis that photoperiodic time measurement in the spider mite is based on a qualitative principle; photoperiods are classified as either 'long' or 'short' in relation to a 'critical' photoperiod. However, around the critical nightlength, intermediate responses were observed which might hint at the quantitative nature of the underlying mechanism. Therefore, although most results are in agreement with the hypothesis of a qualitative mechanism, it cannot be excluded that photoperiodic time measurement in the spider mite is based on a quantitative principle. PMID- 12769963 TI - Control of pupal commitment in the imaginal disks of Precis coenia (Lepidoptera: Nymphalidae). AB - When final (5th) instar larvae of Precis coenia were treated with the juvenile hormone analog (JHA) methoprene, they underwent a supernumerary larval molt, except for certain regions of their imaginal disks, which deposited a normal pupal cuticle. Evidently those regions had already become irreversibly committed to pupal development at the time JHA was applied. By applying JHA at successively later times in the instar, the progression of pupal commitment could be studied. Pupal commitment in the proboscis, antenna, eye, leg and wing imaginal disks occurred in disk-specific patterns. In each imaginal disk there were distinct initiation sites where pupal commitment began during the first few hours of the final larval instar, and from which commitment spread across the remainder of the disk over a 2- to 3-day period. The initiation sites were not always located in homologous regions of the various disks. As a rule, pupal commitment also spread from imaginal disk tissue to surrounding epidermal tissue. The regions of pupal commitment in all disks except those of the wings, coincided with the regions of growth of the disk. Only portions of the disk that had undergone cell division and growth underwent pupal commitment. Shortening the growth period did not prevent pupal commitment in the wing imaginal disk, indicating that, in this disk at least, a normal number of cell divisions was not crucial in reprogramming of disk cells for pupal cuticle synthesis. The apparent growth spurt of imaginal disks that occurs during the last part of the final larval instar is merely the final stage of normal and constant exponential growth. Juvenile hormone (JH) and ecdysteroids appeared to play little role in the regulation of normal imaginal disk growth. Instead, growth of the disks may be under intrinsic control. Interestingly, even though endogenous fluctuation in JH titers do not affect imaginal disk growth, exogenous JHA proved able to inhibit both pupal commitment, cell movement, and growth of the disks during the last larval instar. This function of JH could be important under certain adverse conditions, such as when metamorphosis is delayed in favor of a supernumerary larval molt. PMID- 12769962 TI - Intracellular calcium in prothoracic glands of Manduca sexta. AB - Cytosolic free calcium was measured in individual prothoracic gland cells of Manduca larvae with Fura-2. During the last larval instar there was no correlation between intracellular calcium concentration and ecdysteroid secretion by the glands. The addition of prothoracicotropic hormone (PTTH) from brains of Manduca larvae to prothoracic glands in vitro resulted in a significant increase in the calcium concentration of the gland cells. The effect of PTTH was inhibited by the inorganic calcium channel antagonists, cadmium, lanthanum and nickel, and by the antagonist of T-type calcium channels, amiloride, whereas all the other antagonists tested failed to block the action of PTTH. TMB-8, an inhibitor of intracellular calcium mobilization, did not reduce the PTTH-induced rise in calcium, which suggests that IP(3)-dependent intracellular calcium stores are not involved in the calcium-mediated stimulation of ecdysteroid synthesis. Moreover, PTTH is thought to increase intracellular calcium in prothoracic glands of Manduca by influencing calcium channels in the plasma membrane. PMID- 12769964 TI - A digestive phospholipase A(2) in midguts of tobacco hornworms, Manduca sexta L. AB - We hypothesized that phospholipase A(2) (PLA(2)) is a common feature of insect digestive physiology. PLA(2) hydrolyzes polyunsaturated fatty acids (PUFAs) associated with the sn-2 position of phospholipids (PLs). We describe here a PLA(2) from midgut contents of the tobacco hornworm, Manduca sexta. Our results indicate that the enzyme is sensitive to pH (inactivated at low pH), protein concentration (up to 1.6&mgr;g/&mgr;l), substrate concentration (up to 1.4nmoles/reaction), temperature (up to 30 degrees C), and incubation time. We also found that PLA(2) activity is higher in fed than in starved larvae, and enzyme activity is associated with the midgut contents, rather than the midgut epithelium of fed larvae. All known secretory PLA(2)s, except for a PLA(2) in venom of the marine snail, Conus magus, require high calcium concentrations for catalysis, but the Manduca PLA(2) appears to be calcium-independent, and it exhibits increased PLA(2) activity in the presence of a calcium-chelator, EGTA. In addition, the partially purified Manduca PLA(2) is not inhibited by the phospholipid analog, oleyloxyethylphosphorylcholine. These findings suggest that the Manduca digestive PLA(2) may represent another novel form of PLA(2). PMID- 12769965 TI - Physiological and endocrine changes associated with polydnavirus/venom in the parasitoid-host system Chelonus inanitus-Spodoptera littoralis. AB - As shown earlier, parasitization by the egg-larval parasitoid C. inanitus causes in its host the precocious onset of metamorphosis in the 5th instar followed by developmental arrest in the prepupal stage. Polydnavirus/venom were shown to be responsible for the developmental arrest. We investigated how polydnavirus/venom affect growth of the host larvae and found that head capsule widths were smaller from the 4th to 6th stadium and weights were lower in the 6th stadium in polydnavirus/venom-containing larvae than in non-parasitized larvae. In an attempt to identify endocrine parameters that are modified by polydnavirus/venom and might be responsible for the developmental arrest in the prepupa, we compared juvenile hormones, juvenile hormone esterase and ecdysteroids between non parasitized and polydnavirus/venom-containing larvae from the 4th instar until pupation or developmental arrest, respectively. Obvious differences became manifest only in the 6th instar at the pupal cell formation stage, i.e. 12 days after entry of polydnavirus/venom into the host egg. Then, prothoracic glands of polydnavirus/venom-containing larvae released less ecdysteroids and ecdysteroid titres were lower than in non-parasitized larvae; this was followed by a delayed, reduced and desynchronized increase in prepupal juvenile hormones and juvenile hormone esterase and a slightly modified metabolism of ecdysone. This indicates that polydnavirus/venom affects the endocrine system of the host only after pupal commitment and that inhibition of prothoracic gland activity is the first detectable effect. PMID- 12769966 TI - Effects of parasitization by Cotesia congregata on the brain-prothoracic gland axis of its host, Manduca sexta. AB - The ability of prothoracic glands (PTGs) from parasitized and unparasitized Manduca sexta 5th-instars to respond to ecdysiotropic extracts prepared from day 5 5th instar brains was compared. An in vitro bioassay revealed that PTGs from parasitized animals were much less responsive to brain PTTH than glands from unparasitized larvae. However, when incubated in Grace's medium in the absence of brain extract, glands from day-3 and -4 hosts remained active for a much longer period of time than did those dissected from their unparasitized counterparts. Rather than exhibiting reduced (basal) levels of synthesis after the 3rd hour of incubation, glands from these parasitized larvae continued to synthesize/release ecdysteroid into the medium at relatively high rates. The timing of this enhanced secretory activity is coincident with the ecdysteroid peak that occurs just prior to and during wasp emergence. Following parasite emergence, gland activity decreased, and by the third day after emergence, was reduced to low levels. Results suggest that the requirement for PTTH to stimulate ecdysteroid production has been bypassed, i.e. that the parasite has uncoupled the normal mechanisms that permit brain regulation of PTG activity. The ability of brains from parasitized M. sexta to stimulate PTGs from unparasitized day-2 5th instars was also examined. Dose-response analyses performed for the first 7 days of the 5th instar showed that on a per brain basis ecdysiotropic activity in brains from parasitized and unparasitized animals was similar. However, when differences in brain size were considered, ecdysiotropic activity appeared to be more concentrated in brains from day-7 parasitized larvae than in brains from similarly aged unparasitized larvae. Analysis of the size distribution of the ecdysiotropic activity in brains from parasitized larvae revealed a unique form that was larger than the 29kDa standard. This suggests that parasitization may inhibit neuropeptide processing, particularly during the final stages preceding emergence of the wasps from the host. Thus, both an inhibition of prothoracicotropic hormone processing and the inability to respond to this neurohormone may contribute to the developmental arrest characteristic of parasitized 5th instars. PMID- 12769967 TI - Phe-X-Pro-Arg-Leu-NH(2) peptide producing cells in the central nervous system of the silkworm, Bombyx mori. AB - Members of the neuropeptide family having Phe-X-Pro-Arg-Leu-NH(2) (FXPRLamide; X=Ser, Thr, Val, or Gly) at the C-terminus serve as regulators of oviduct and visceral muscle contraction, sex pheromone production, and diapause induction. Antibody raised against Bombyx mori diapause hormone recognized a variety of FXPRLamide peptides. Using this antibody, the antigen was immunocytochemically localized in the central nervous system (CNS) of the silkworm, Bombyx mori. Immunoreactive somata were observed in all ganglia of the CNS including the brain. Twelve somata localized at the midline of the suboesophageal ganglion (SG) were most intensely stained, and their neurite projections reached the retrocerebral complex. Thus, these cells in the SG exhibited typical features of neuroendocrine neurons. Marked reduction in immunoreactivity was observed in a pair of neurosecretory cells in the labial neuromere in SG of diapause type pupae, which indicates an active release of FXPRLamide peptides from these cells. No clear connection to neurohemal sites were observed in immunoreactive cells in the brain, thoracic or abdominal ganglia, suggesting that the immunoreactive peptides in these organs are likely to serve as neurotransmitters or neuromodulators. PMID- 12769968 TI - Spatial coincidence of cues in visual learning by the honeybee (Apis mellifera). AB - The discrimination of patterns was studied in a Y-choice chamber fitted with a transparent baffle in each arm, through which the bees had a choice of two targets via openings 5cm wide. The bees see the positive (rewarded) and the negative (unrewarded) targets from a fixed distance. The patterns were bars (subtending 22 degrees x5.4 degrees at the point of choice) presented in one quarter of each target. The bars were moved to a different quarter of the target every 5min, to make the location of black useless as a cue. A coincident presentation is when the bar on the left target is on the same side of the target as the bar on the right target. The bees learn the orientation cue when the presentation is coincident but otherwise cannot learn it. This experiment shows that bees do not centre their attention on the individual bars, otherwise they would always discriminate the orientation. Centring the target as a whole precedes learning. Having learned with the bar on one side of the targets, bees do not recognize the same cue presented on the other side. A separate orientation cue can be learned on each side. A radial/tangential cue is preferred to a conflicting orientation cue. PMID- 12769969 TI - Offspring and sex ratio are independent of sperm management in Eupelmus orientalis females. AB - Sperm stocks in both males and females of the parthenogenetic wasp Eupelmus orientalis were investigated at various points during reproduction and compared to the progeny of females in controlled conditions. One day-old virgin males had approximately 5500 sperm, and from a total of about 1697 sperm transferred per copulation, 21% are stored in the spermathecae by females 24 hours after mating. At the end of the egg-laying period (at least 42 days), 2/5 of the initial amount of sperm remained in this storage organ. This decrease (from approximately 350 to 150) occurred essentially during the first 21 days of egg-laying activity, indicating that the majority of sperm stored were used during this period. Between 21 days and the end of fertile life, the number of sperm remained constant. The mean offspring production throughout reproductive life after one mating was 153, with 56.5% of the daughters laid at the beginning of the laying activity. Sex ratio was entirely female biased during the first 15 days (mean=0.65), then it decreased and became nearly equal after 20 days. Present results propose that females maximize the production of daughters i.e. of inseminated eggs until the 20th day and after this time lay as many daughters and sons despite their still having stored sperm. Physiological constraints due to ageing are proposed to explain this phenomenon. PMID- 12769970 TI - Utilisation of prey by antlion larvae (Neuroptera: Myrmeleontidae) in terms of energy and nutrients. AB - Prey utilisation at low prey densities was determined for third instar Cueta sp., Furgella intermedia (Markl) and Palpares annulatus (Stitz) larvae in terms of wet weight, dry weight, energy and nutrients. Prey utilisation was similar to other insects on a wet weight (42-47%), dry weight (46-49%), energetic (40-58%) and nutritive basis (62-79%). Lipids (33-36%) provided energetically the highest contribution of the nutrients ingested. The quantities of water, proteins, nucleic acids, lipids and carbohydrates extracted by the antlion larvae were in proportion to their availability in their prey, the Hodotermes mossambicus larvae. The quantities of nutrients extracted by the antlion larvae at low prey densities were not significantly influenced by the differences in mandible size, antlion body weight or the trapping method (building a pit or not) of the antlion species. It is proposed that a low metabolic rate and the accumulation of fat reserves, and not the extent of prey utilisation, enable P. annulatus larvae to tolerate a 123-d starvation period in which 22.3% of their body weight is lost. PMID- 12769971 TI - Melatonin and the pea aphid, Acyrthosiphon pisum. AB - Pea aphids, Acyrthosiphon pisum, were fed on artificial diet containing various concentrations of melatonin. Under long-day conditions (16h light:8h dark) their progeny included males and virginoparous/oviparous (asexual/sexual) intermediate females, which normally occur only in short days or around critical night-length. Endogenous melatonin in pea aphids was measured by radioimmunoassay and verified by parallelism with a melatonin standard curve and by thin layer chromatography. However, melatonin titres showed large variations and although they tended to be higher during the scotophase than during the photophase they were not significantly different. The possibility of melatonin being involved in photoperiodism is discussed. PMID- 12769972 TI - Avoidance of intracellular freezing by the freezing-tolerant New Zealand Alpine weta Hemideina maori (Orthoptera: Stenopelmatidae). AB - Calorimetric analysis indicates that 82% of the body water of Hemideina maori is converted into ice at 10 degrees C. This is a high proportion and led us to investigate whether intracellular freezing occurs in H. maori tissue. Malpighian tubules and fat bodies were frozen in haemolymph on a microscope cold stage. No fat body cells, and 2% of Malpighian tubule cells froze during cooling to -8 degrees C. Unfrozen cells appeared shrunken after ice formed in the extracellular medium. There was no difference between the survival of control tissues and those frozen to -8 degrees C. At temperatures below -15 degrees C (lethal temperatures for weta), there was a decline in survival, which was strongly correlated with temperature, but no change in the appearance of tissue. It is concluded that intracellular freezing is avoided by Hemideina maori through osmotic dehydration and freeze concentration effects, but the reasons for low temperature mortality remain unclear. The freezing process in H. maori appears to rely on extracellular ice nucleation, possibly with the aid of an ice nucleating protein, to osmotically dehydrate the cells and avoid intracellular freezing. The lower lethal temperature of H. maori (-10 degrees C) is high compared to organisms that survive intracellular freezing. This suggests that the category of 'freezing tolerance' is an oversimplification, and that it may encompass at least two strategies: intracellular freezing tolerance and avoidance. PMID- 12769973 TI - Structure-activity studies reveal two allatostatin receptor types in corpora allata of Diploptera punctata. AB - Synthetic variants of the octadecapeptide amide ASB2 (AYSYVSEYKRLPVYNFGL-NH(2)), a cockroach allatostatin, were assayed in vitro on corpora allata (CA) from 2-day old (vitellogenic) and 10-day-old (post-vitellogenic) female Diploptera punctata. The analogs [(17)psi(18),CH(2)-S]ASB2, [D-Trp(17)]ASB2 and [Ile(18)]ASB2 inhibited juvenile hormone (JH) synthesis with simple dose-response curves on sensitive CA from 10-day-old females. These analogs were fully effective but less potent than ASB2. When tested on CA from 2-day-old mated females, which are only partially (65-70%) sensitive to ASB2, the three analogs gave biphasic dose response curves and elicited a maximal effect only at higher concentrations. The dose-response curve for ASB2 on CA from 2-day-old females had a Hill plot slope of only 0.78+/-0.03. These findings suggested that the observed CA sensitivity to ASB2 may be the result of two partial responses having an IC(50) of approximately 0.35 and 3nM respectively. One partial response, or receptor type, appeared more sensitive than the other to adverse modification of the "message" segment of the peptide. The activity of shorter allatostatins was also studied, indicating that pentapeptides of the YXFGL-amide structure are fully effective, albeit at low potency, as inhibitors of JH biosynthesis. PMID- 12769974 TI - An allatostatic factor and juvenile hormone synthesis by corpora allata in Locusta migratoria. AB - The hemolymph juvenile hormone (JH) titer in sexually immature female adults of Locusta migratoria (Ibaraki strain, Japan) was lower than in sexually mature females; nevertheless, JH synthetic activity by the corpora allata (CA) in vitro was considerably higher in immature females than in sexually mature females ([Okuda et al., 1996]). We carried out experiments to explain this contradiction. The CA activity of sexually immature female adults was very low when the CA were incubated as a complex together with the corpora cardiaca (CC) and brain. When the same complex was assayed after cutting the nerve cord connecting the CC and CA (NCA1), JH synthesis by the CA was enhanced tenfold. When this pair of CA was incubated in fresh medium without the CC and brain, JH synthesis was further increased. Therefore, the higher in-vitro JH production by CA from immature female adults was the result of isolation of the CA from the brain and CC. A methanolic extract of brain-CC complexes contained a factor that inhibited JH synthetic activity by CA in vitro in both immature and mature insects, and this inhibition was reversible. The factor was heat-resistant but lost allatostatic activity after pronase digestion. These results indicate that the allatostatic factor is probably a heat-stable peptide. PMID- 12769975 TI - Ecdysone 20-hydroxylation and 3-epimerization in larvae of the gypsy moth, Lymantria dispar L.: tissue distribution and developmental changes. AB - The potential for ecdysone metabolism was determined for various larval tissues of the gypsy moth, Lymantria dispar. Homogenates of fat body, midguts, and Malpighian tubules, taken on different days during the second half of the fifth instar, were incubated with [(3)H]ecdysone, and the products were analyzed by reversed-phase and normal-phase HPLC. All tissues showed conversion to 20 hydroxyecdysone, and midguts also produced 3-epiecdysone. Ecdysone 20 monooxygenase (E20MO) activity in the fat body increased from a low level on day 5 to a peak on day 11, coinciding with the peak in the hemolymph ecdysteroid titer on the penultimate day of the instar. Midguts and Malpighian tubules showed E20MO activity only during the last 3 or 4days of the instar, with the highest activity also occurring on the penultimate day. For the midguts, the appearance of the E20MO coincided with the transition from larval to pupal tissue. No activity was detected in larval midguts. 3-Epiecdysone formation, however, was mainly found in larval midguts, with only marginal activity detectable in pupal midguts. PMID- 12769976 TI - Ecdysteroidogenic action of Bombyx prothoracicotropic hormone and bombyxin on the prothoracic glands of Rhodnius prolixus in vitro. AB - An in-vitro assay for ecdysteroid synthesis by the prothoracic glands (PGs) of fifth instar Rhodnius prolixus has been employed to evaluate the actions of prothoracicotropic neuropeptides from the silkmoth, Bombyx mori. Crude prothoracicotropic hormone (PTTH) extracts of recently emerged adult brain complexes of Bombyx induced a dose-dependent stimulation of ecdysteroid synthesis by Rhodnius PGs, which was similar to that obtained using crude Rhodnius PTTH. In both cases, maximum stimulation was obtained with one brain equivalent. Rhodnius PGs were then challenged with incremental doses of recombinant Bombyx PTTH and synthetic bombyxin-II. Dose-response curves for the action of both peptides on Rhodnius PGs were very similar to those obtained for their action on the pupal PGs of Bombyx in vitro. Bombyx PTTH stimulated the PGs of Rhodnius at concentrations comparable to those effective on Bombyx. The curve for Bombyx PTTH showed a steep ascending region from 3 to 8ng/ml and a sharp peak. For bombyxin, concentrations 40-fold higher were required to elicit the same amount of stimulation as obtained using Bombyx PTTH. Therefore, Rhodnius PGs possess recognition sites for both Bombyx PTTH and bombyxin. This is the first study of the ecdysteroidogenic properties of the Bombyx peptides on a heterologous species. It is suggested that the function and conformation of PTTH may be conserved between distantly related insect groups. PMID- 12769977 TI - KCl reabsorption by the lower malpighian tubule of rhodnius prolixus: inhibition by Cl(-) channel blockers and acetazolamide. AB - Iono- and osmoregulation by the blood-feeding hemipteran Rhodnius prolixus involves co-ordinated actions of the upper and lower Malpighian tubules. The upper tubule secretes ions (Na(+), K(+), Cl(-)) and water, whereas the lower tubule reabsorbs K(+) and Cl(-) but not water. The extent of KCl reabsorption by the lower tubule in vitro was monitored by ion-selective microelectrode measurement of Cl(-) and/or K(+) concentration in droplets of fluid secreted by Malpighian tubules isolated under oil. An earlier study proposed that K(+) reabsorption involves an omeprazole-sensitive apical K(+)/H(+) ATPase and Ba(2+) sensitive basolateral K(+) channels. This paper examines the effects acetazolamide and of compounds that inhibit chloride channels, Cl(-)/HCO(3)(-) exchangers and Na(+)/K(+)/2Cl(-) or K(+)/Cl(-) co-transporters. The results suggest that Cl(-) reabsorption is inhibited by acetazolamide and by Cl(-) channel blockers, including diphenylamine-2-carboxylate(DPC) and 5-nitro-2-(3 phenylpropylamino) benzoic acid (NPPB), but not by compounds that block Na(+)/K(+)/Cl(-) and K(+)/Cl(-) co-transporters. Measurements of transepithelial potential and basolateral membrane potential during changes in bathing saline chloride concentration indicate the presence of DPC- and NPPB-sensitive chloride channels in the basolateral membrane. A working hypothesis of ion movements during KCl reabsorption proposes that Cl(-) moves from lumen to cell through a stilbene-insensitive Cl(-)/HCO(3)(-) exchanger and then exits the cell through basolateral Cl(-) channels. PMID- 12769978 TI - Helix pomatia lectin and annexin V, two molecular probes for insect microparticles: possible involvement in hemolymph coagulation. AB - Insect hemolymph coagulation involves a complex reaction with contributions from hemocytes and soluble factors. Here we present evidence for the presence of microparticles in the coagulation reaction. These particles are formed by hemocytes in a calcium-dependent process. Both the particles and the remaining cells are labelled by annexin V indicating the presence of phosphatidylserine on the outer membrane. Microparticles are enriched in hemomucin, a surface protein of Drosophila hemocytes that is specifically recognised by a snail (Helix pomatia) lectin. Hemomucin is shown to bind to lipophorin, a multifunctional hemolymph molecule previously implied in coagulation. Our findings suggest similarities at a biochemical and cellular level between vertebrate blood and insect hemolymph coagulation. PMID- 12769979 TI - cAMP-stimulated termination of vitellogenesis in Hyalophora cecropia: formation of a diffusion barrier and the loss of patency. AB - Activation of cAMP-dependent protein kinase (PKA) by cell-permeable analogs of cAMP causes early and mid-vitellogenic follicles of Hyalophora cecropia to terminate vitellogenin uptake [[Wang and Telfer, 1996], Insect Biochem. Mol. Biol. 26, 85-94 (1996)]. The response is shown here to entail the formation of an epithelial diffusion barrier. Follicle cells that have been loosely organized to provide intercellular pathways for the movement of vitellogenin to the oocyte surface transform into a tight epithelium within 1-2h of exposure to PKA activators. The follicle cells can now prevent the escape of Lucifer yellow CH that has been iontophoresed into the space surrounding the oocyte, and the entry of labeled vitellogenin from the medium. As they form this functional equivalent of a tight junction, the follicle cells further reduce the intercellular spaces by enlarging and pressing against each other, and by slowing the secretion of the sulfated glycosaminoglycan matrix that separates them during vitellogenesis. The activation of PKA in early and mid-vitellogenic follicles thus appears to trigger prematurely a set of changes that do not normally occur until the follicle has grown to a length of about 2.0mm. PMID- 12769980 TI - Critical thermal limits, temperature tolerance and water balance of a sub Antarctic caterpillar, Pringleophaga marioni (Lepidoptera: Tineidae). AB - Thermal tolerance, supercooling point, water balance and osmoregulatory ability of Pringleophaga marioni Viette (Lepidoptera: Tineidae) are investigated in this study. Field-fresh larvae had a mean CT(Min) (cold stupor) of -0.6 degrees C and a mean CT(Max) (heat coma) of 38.7 degrees C. The mean supercooling point of field-fresh individuals was -5.0 degrees C. Caterpillars showed 100% survival of freezing to -6.5 degrees C, but at -12 degrees C mortality rose to 100%. Survival of a 30h exposure to -6.0 degrees C was 80%, but declined to 30% in the 6-12h interval at -7.5 degrees C. No caterpillars survived for longer than 12h at -9.0 degrees C. Survival of high temperatures (35 degrees C and above) was poor. Tolerance of water loss (46% of fresh mass) and rates of water loss (1% fresh massh(-1)) were similar to those found in other mesic insects. P. marioni larvae were incapable of metabolizing lipids to replenish lost water and showed no haemolymph osmoregulatory ability. It is suggested that the preponderance of freeze tolerance in high-latitude southern hemisphere species may be associated with their occurrence in moist habitats, and that the "freeze tolerance" category be re-examined in the light of the range of strategies adopted by such arthropods. PMID- 12769981 TI - Does PBAN play an alternative role of controlling pheromone emission in the cabbage looper moth, Trichoplusia ni (Hubner) (Lepidoptera: Noctuidae)? AB - There is an active process by which sex pheromone reserves of female cabbage looper moths, Trichoplusia ni, are transported to the gland's surface during the nocturnal period of calling. We hypothesized that this mobilization was controlled by a head factor, possibly related to the pheromone biosynthesis activating neuropeptides (PBAN) that in other species stimulate pheromone synthesis. We evaluated the impact of head extracts of T. ni on pheromone emission and glandular content of pheromone. During the photophase injected head extracts stimulated an increased pheromone emission rate in females, but glandular content of pheromone was not affected. Head extracts of H. virescens, a species with known PBAN activity, and synthetic PBAN stimulated an increased pheromone emission rate in T. ni. There was some specificity of the response of female T. ni to PBAN, in that several other unrelated polypeptides did not stimulate this type of response. Previously it had been determined that brain factors do not play a role in stimulating pheromone biosynthesis in T. ni. Our results indicate that there may be additional avenues by which PBAN or related neuropeptides control pheromone emission, including transport of pheromone reserves to the surface of the sex pheromone gland. PMID- 12769982 TI - Quantification and visualization of Dippu-AST mRNA in the brain of adult Diploptera punctata: mated females vs. virgin females vs. males. AB - YXFGLa-allatostatins (ASTs) are a family of neuropeptides that inhibit juvenile hormone biosynthesis in vitro by the corpora allata of cockroaches and crickets. We have investigated the levels of AST expression in the brain of adult mated and virgin females, and adult males of the cockroach Diploptera punctata, for 11 days following emergence. To quantify Dippu-AST mRNA we have used an optimized, sensitive and specific quantitative competitive reverse-transcriptase polymerase chain reaction (QC-RT-PCR) method. Using this technique, we show that Dippu-AST is expressed at higher levels in the brains of mated females than in the brains of virgin females. Specifically, levels of Dippu-AST mRNA in day 0 mated females are 2.5x higher than levels in day 0 virgin females. By day 6-7, levels in mated females are 20x higher than those in virgin females. However, the general pattern of expression is preserved in mated and virgin females; levels of Dippu-AST mRNA are relatively high in newly eclosed females, fall steadily to significantly lower levels on day 5 (mated) and day 5-7 (virgins), and then increase and remain relatively stable until at least day 10. No significant changes in Dippu-AST mRNA are apparent in male brains during the observation period. However, the general pattern of expression is virtually the inverse of that seen in females. Dippu-AST mRNA in male brains is relatively low following emergence, increases to maximum levels on day 5, following which, levels steadily fall. In situ hybridization suggests that fluctuations in Dippu-AST mRNA expression are a result of differential transcription within medial and lateral cells of the pars intercerebralis in both males and females. In addition to yielding insights into Dippu-AST function, the quantification of Dippu-AST expression under normal conditions is useful in providing a basis for comparison of Dippu-AST expression following various physiological manipulations. PMID- 12769983 TI - Effects of dietary variation on growth, composition, and maturation of Manduca sexta (Sphingidae: Lepidoptera). AB - Most studies linking dietary variation with insect fitness focus on a single dietary component and late larval growth. We examined the effects of variation in multiple dietary factors over most life stages of the sphingid moth, Manduca sexta. Larvae received artificial diets in which protein, sucrose, and water content were varied. The relationship between larval size, growth and consumption rates differed significantly across diets. Larvae on control and low-sucrose diets grew most rapidly and attained the largest pupal and adult sizes. Conversely, larvae on low-water and low-protein diets initially grew slowly, but accelerated in the fifth instar and became pupae and adults comparable to control animals in size. There were no fundamental differences in protein:carbohydrate consumption patterns or strategies among experimental diets and larval instars. However, inadequate dietary water appeared to be more important for early than late instar larvae. Larvae on all artificial diets showed increasing fat content throughout all stages, including wandering and metamorphosis. Compensatory feeding among low-water and low-protein larvae was correlated with significantly higher fat content in larvae, pupae and adults, whereas low-sucrose animals were substantially leaner than those on the control diet. These differences may have strong effects on adult physiology, reproduction, and foraging patterns. PMID- 12769984 TI - Survival of Atta sexdens workers on different food sources. AB - Leaf-cutting ants belonging to the tribe Attini are major herbivores and important agriculture pests in the neotropics, these ants being thought to feed on the sap which exudes from the plant material which they cut and also on the mycelium of a symbiotic fungus that grows on plant material inside their nests in what is called "the fungus garden". However, we have found that the survival of Atta sexdens worker ants on leaves, on mycelium of the ants' symbiotic fungus, Leucoagaricus gongylophorus, or on plant polysaccharides was the same as that of starved A. sexdens, while, conversely, significantly longer survival was achieved by ants fed on the fungus garden material or on some of the products (especially glucose) of the hydrolysis of plant polysaccharides. We found that the fungus garden contained glucose at a higher concentration than that found in leaves or fungal mycelium, and that this glucose was consumed by the ant to the extent that it was probably responsible for up to 50% of the nutritional needs of the workers. The fungus garden contained polysaccharide degrading enzymes (pectinase, amylase, xylanase and cellulase) in proportions similar to that observed in laboratory cultures of L. gongylophorus. It thus appears that A. sexdens workers obtain a significant part of their nutrients from plant polysaccharide hydrolysis products produced by the action of extracellular enzymes released by L. gongylophorus. In this paper we discuss the symbiotic nutrition strategy of A. sexdens workers and brood and the role played by plant polysaccharides in the nutrition of attine ants. PMID- 12769985 TI - The role of water vapour in the orientation behaviour of the blood-sucking bug Triatoma infestans (Hemiptera, Reduviidae). AB - The behavioural response to water vapour of the haematophagous bug Triatoma infestans was analysed. Dry or humid discrete sources at different temperatures were used as stimuli for insects walking on a locomotion compensator. Humidity significantly increased the tendency of these bugs to orientate towards thermal sources. Furthermore, humid sources at room temperature were attractive to T. infestans, but this effect was limited to short-range distances. On the other hand, dynamic sources, i.e. airstreams carrying different water vapour contents did not affect the spontaneous anemotactic behaviour of this species, neither in sign (positive) nor in intensity. The anemotactic behaviour was also not influenced by the physiological water balance state of the bugs. Results are discussed in relation to the cues released by living hosts of triatomine bugs and in relation to their responses to air-currents. PMID- 12769986 TI - Myotropic effect of helicokinins, tachykinin-related peptides and Manduca sexta allatotropin on the gut of Heliothis virescens (Lepidoptera: Noctuidae). AB - Different insect neuropeptides (helicokinins, tachykinin-related and allatoregulating peptides) were investigated with regard to their myostimulatory effects using whole-gut preparations isolated from fifth instar Heliothis virescens larvae. The experiments demonstrated that representatives of all three peptide families are able to induce and amplify gut contractions in this species in a dose-dependent manner. Structure-activity studies (alanine scan, D-amino acid scan and truncated analogues) with the helicokinin Hez-K1 supported the finding, that the core sequence for biological activity of kinins is the amidated C-terminal pentapeptide (FSPWG-amide). Similar investigations with insect tachykinin isolated from Leucophaea madera (Lem-TRP1) revealed that the minimum sequence evoking a physiological gut response in H. virescens is the amidated hexapeptide (GFLGVR-amide), which represents the conserved amino acid sequence for Leucophaea TRPs in general. The peptide concentration causing a half-maximal gut contraction (EC(50)) for Lem-TRP1 was about 26 nM. Although the potency of Lem-TRP1 was 9-fold lower compared with Hez-KI (EC(50): 3 nM), the maximal tension of the gut obtained with Lem-TRP1 was 1.7-fold higher compared with Hez KI. The EC(50) of Manduca sexta allatotropin (Mas-AT; 79 nM) was of lowest potency among all three peptides tested. In a pharmacological study, co incubation experiments with Lem-TRP1, Hez-KI or Mas-AT and compounds interfering with signal transduction pathways were employed to investigate the mode of action of the myotropic effects of these peptides. Cadmium and the protein kinase C (PKC) inhibitor tamoxifen attenuated the contractile effects of all three peptides tested. The data suggest that in the gut muscle of H. virescens the myotropic peptides bind to G-protein-coupled receptors that cause contraction by promoting the entry of extracellular calcium mediated by a PKC involved pathway. PMID- 12769987 TI - The role of the subelytral cavity in respiration in a tenebrionid beetle, Onymacris multistriata (Tenebrionidae: Adesmiini). AB - This study measured the respiratory patterns in the tenebrionid beetle, Onymacris multistriata, using flow-through respirometry to measure carbon dioxide emission from the mesothoracic spiracles separately and simultaneously with that from around the elytral case. 96% of the total CO(2) emitted was via the mesothoracic spiracles. These spiracles used a discontinuous gas exchange cycle similar to that measured from other tenebrionid beetles. Although the circadian rhythm of the beetles resulted in changes to the period durations and cycle frequencies in the discontinuous gas exchange cycles, the mesothoracic spiracle remained the major site for gas exchange. Thus the subelytral cavity plays a different role in respiration other than the elimination of CO(2) build-up. It is expected that other arid dwelling flightless beetles will also be shown to use the mesothoracic spiracle as the major route for CO(2) emission. PMID- 12769988 TI - Fluid intake rates in ants correlate with their feeding habits. AB - This study investigates the techniques of nectar feeding in 11 different ant species, and quantitatively compares fluid intake rates over a wide range of nectar concentrations in four species that largely differ in their feeding habits. Ants were observed to employ two different techniques for liquid food intake, in which the glossa works either as a passive duct-like structure (sucking), or as an up- and downwards moving shovel (licking). The technique employed for collecting fluids at ad libitum food sources was observed to be species-specific and to correlate with the presence or absence of a well developed crop in the species under scrutiny. Workers of ponerine ants licked fluid food during foraging and transported it as a droplet between their mandibles, whereas workers of species belonging to phylogenetically more advanced subfamilies, with a crop capable of storing liquids, sucked the fluid food, such as formicine ants of the genus Camponotus. In order to evaluate the performance of fluid collection during foraging, intake rates for sucrose solutions of different concentrations were measured in four ant species that differ in their foraging ecology. Scaling functions between fluid intake rates and ant size were first established for the polymorphic species, so as to compare ants of different size across species. Results showed that fluid intake rate depended, as expected and previously reported in the literature, on sugar concentration and the associated fluid viscosity. It also depended on both the species-specific feeding technique and the extent of specialization on foraging on liquid food. For similarly-sized ants, workers of two nectar-feeding ant species, Camponotus rufipes (Formicinae) and Pachycondyla villosa (Ponerinae), collected fluids with the highest intake rates, while workers of the leaf-cutting ant Atta sexdens (Myrmicinae) and a predatory ant from the Rhytidoponera impressa-complex (Ponerinae) did so with the lowest rate. Calculating the energy intake rates in mg sucrose per unit time, licking was shown to be a more advantageous technique at higher sugar concentrations than sucking, whereas sucking provided a higher energy intake rate at lower sugar concentrations. PMID- 12769989 TI - Larval juvenile hormone treatment affects pre-adult development, but not adult age at onset of foraging in worker honey bees (Apis mellifera). AB - Previous research has shown that juvenile hormone (JH) titers increase as adult worker honey bees age and treatments with JH, JH analogs and JH mimics induce precocious foraging. Larvae from genotypes exhibiting faster adult behavioral development had significantly higher levels of juvenile hormone during the 2nd and 3rd larval instar. It is known that highly increased JH during this period causes the totipotent female larvae to differentiate into a queen. We treated third instar larvae with JH to test the hypothesis that this time period may be a developmental critical period for organizational effects of JH on brain and behavior also in the worker caste, such that JH treatment at a lower level than required to produce queens will speed adult behavioral development in workers. Larval JH treatment did not influence adult worker behavioral development. However, it made pre-adult development more queen-like in two ways: treated larvae were capped sooner by adult bees, and emerged from pupation earlier. These results suggest that some aspects of honey bee behavioral development may be relatively insensitive to pre-adult perturbation. These results also suggest JH titer may be connected to cues perceived by the adult bees indicating larval readiness for pupation resulting in adult bee cell capping behavior. PMID- 12769990 TI - Passive evasion of encapsulation in Macrocentrus cingulum Brischke (Hymenoptera: Braconidae), a polyembryonic parasitoid of Ostrinia furnacalis Guenee (Lepidoptera: Pyralidae). AB - The hymenopteran Macrocentrus cingulum usually deposits one egg into the larval body cavity of lepidopteran Ostrinia furnacalis, and the egg subsequently splits into several dozens of embryos during its development. How the parasitoid eggs and embryos avoid encapsulation by the host's immune response remains unknown. We compared hemocyte counts, morphologies and behaviors between unparasitized O. furnacalis larvae, and larvae parasitized by M. cingulum. No distinct differences were observed. Sephadex A-25 beads elicited a strong encapsulation response when injected into the parasitized host larvae, which indicates that parasitism by M. cingulum does not affect host's cellular immunity. However, there were significant differences in the host's encapsulation reactions towards injected eggs from different sources. Injected M. cingulum mature eggs excised from the lateral oviducts of the female wasps were not encapsulated, while immature eggs or driselase treated mature ones provoked an encapsulation response within 2 h after injection. Inspection of eggs by transmission electron microscopy revealed that the driselase collapsed the surface fibrous layer of the eggs, indicating that surface fibrous layer may play a role in protecting eggs from host's immune attack. PMID- 12769991 TI - Structural and biochemical analysis of the Leptinotarsa decemlineata (Coleoptera; Chrysomeloidea) crystalline chorionic layer. AB - The developmental aspects of the Leptinotarsa decemlineata crystalline chorionic layer (CCL) morphogenesis, its composition and its supramolecular structure were studied. The mature Leptinotarsa decemlineata eggshell consists of the vitelline membrane and the CCL, while the follicle cell remnants following their degeneration after oogenesis completion constitute the outer chorionic layer. The vitelline membrane and the CCL layers are formed through continuous material deposition from the follicular epithelium, whereas the main morphogenic factor during most insect eggshell formation, namely the follicle cell and oocyte microvilli, are seemingly involved only in vitelline membrane formation. Analysis of the CCL morphogenesis showed that this layer is assembled from a fiber-like pre-crystalline material, which accumulates at the vitelline membrane-follicle cell interface. The mature CCL is about 1 microm thick and exhibits a periodicity of approximately 10 nm, while computer image analysis studies of thin-sectioned CCL revealed the existence of crystalline layers parallel to the CCL surface. Finally, SDS-PAGE-electrophoresis of purified CCLs showed that this crystalline layer is of a proteinaceous nature and is most likely composed of 3-5 polypeptides with a molecular weight ranging in between 28-60 kDa. Overall, these data exemplify for the first time the nature and supramolecular arrangement of a crystalline layer and its constituent molecules in Coleoptera. PMID- 12769992 TI - Acquisition of freezing tolerance in early autumn and seasonal changes in gall water content influence inoculative freezing of gall fly larvae, Eurosta solidaginis (Diptera, Tephritidae). AB - We examined seasonal changes in freeze tolerance and the susceptibility of larvae of the gall fly, Eurosta solidaginis to inoculative freezing within the goldenrod gall (Solidago sp.). In late September, when the water content of the galls was high (approximately 55%), more than half of the larvae froze within their galls when held at -2.5 degrees C for 24 h, and nearly all larvae froze at -4 or -6 degrees C. At this time, most larvae survived freezing at > or = -4 degrees C. By October plants had senesced, and their water content had decreased to 33%. Correspondingly, the number of larvae that froze by inoculation at -4 and -6 degrees C also decreased, however the proportion of larvae that survived freezing increased markedly. Gall water content reached its lowest value (10%) in November, when few larvae froze during exposure to subzero temperatures > or = -6 degrees C. In winter, rain and melting snow transiently increased gall water content to values as high as 64% causing many larvae to freeze when exposed to temperatures as high as -4 degrees C. However, in the absence of precipitation, gall tissues dried and, as before, larvae were not likely to freeze by inoculation. Consequently, in nature larvae freeze earlier in the autumn and/or at higher temperatures than would be predicted based on the temperature of crystallization (T(c)) of isolated larvae. However, even in early September when environmental temperatures are relatively high, larvae exhibited limited levels of freezing tolerance sufficient to protect them if they did freeze. PMID- 12769993 TI - Characterisation of the feeding behaviour of western flower thrips in terms of electrical penetration graph (EPG) waveforms. AB - Western flower thrips, Frankliniella occidentalis (Pergande) causes damage to plants when they are feeding. Also, this thrips species transmits Tomato spotted wilt virus (TSWV) during stylet penetration. We investigated the penetration behaviour (probing) of thrips on pepper leaves and on liquid diet by electrical penetration graph (EPG, DC-system) recording. In addition, we used high magnification video observations to correlate EPG waveforms with the insect's posture, head movements, and muscle contractions. Also, EPGs were correlated with probing on liquid diets containing radio-active tracers to distinguish and quantify ingestion waveforms. The previously described waveforms P, Q, and R were distinguished and additionally, a new waveform 'S' was distinguished. Waveform P could be linked with mandibular leaf penetration, waveform Q presumably with insertion of the maxillary stylets, and waveform R with ingestion of cell contents, whereas waveform S could not be correlated with any behavioural activity. Histology of the feeding damage in pepper leaves shows that thrips ingests the contents of multiple cells per probe. PMID- 12769994 TI - Desiccation stress at sub-zero temperatures in polar terrestrial arthropods. AB - Cold tolerant polar terrestrial arthropods have evolved a range of survival strategies which enable them to survive the most extreme environmental conditions (cold and drought) they are likely to encounter. Some species are classified as being freeze tolerant but the majority of those found in the Antarctic survive sub-zero temperatures by avoiding freezing by supercooling. For many arthropods, not just polar species, survival of desiccating conditions is equally important to survival of low temperatures. At sub-zero temperatures freeze avoiding arthropods are susceptible to desiccation and may lose water due to a vapour diffusion gradient between their supercooled body fluids and ice in their surroundings. This process ceases once the body fluids are frozen and so is not a problem for freeze tolerant species. This paper compares five polar arthropods, which have evolved different low temperature survival strategies, and the effects of exposure to sub-zero temperatures on their supercooling points (SCP) and water contents. The Antarctic oribatid mite (Alaskozetes antarcticus) reduced its supercooling point temperature from -6 to -30 degrees C, when exposed to decreasing sub-zero temperatures (cooled from 5 to -10 degrees C over 42 days) with little loss of body water during that period. However, Cryptopygus antarcticus, a springtail which occupies similar habitats in the Antarctic, showed a decrease in both water content and supercooling ability when exposed to the same experimental protocol. Both these Antarctic arthropods have evolved a freeze avoiding survival strategy. The Arctic springtail (Onychiurus arcticus), which is also freeze avoiding, dehydrated (from 2.4 to 0.7 g water g(-1) dry weight) at sub-zero temperatures and its SCP was lowered from c. -3 to below -15 degrees C in direct response to temperature (5 to -5.5 degrees C). In contrast, the freeze tolerant larvae of an Arctic fly (Heleomyza borealis) froze at c. -7 degrees C with little change in water content or SCP during further cold exposure and survived frozen to -60 degrees C. The partially freeze tolerant sub-Antarctic beetle Hydromedion sparsutum froze at c. -2 degrees C and is known to survive frozen to -8 degrees C. During the sub-zero temperature treatment, its water content reduced until it froze and then remained constant. The survival strategies of such freeze tolerant and freeze avoiding arthropods are discussed in relation to desiccation at sub-zero temperatures and the evolution of strategies of cold tolerance. PMID- 12769995 TI - Saliva secretion and ionic composition of saliva in the cockroach Periplaneta americana after serotonin and dopamine stimulation, and effects of ouabain and bumetamide. AB - Isolated salivary glands of Periplaneta americana were used to measure secretion rates and, by quantitative capillary electrophoresis, Na(+), K(+), and Cl(-) concentrations in saliva collected during dopamine (1 micro M) and serotonin (1 micro M) stimulation in the absence and presence of ouabain (100 micro M) or bumetanide (10 micro M). Dopamine stimulated secretion of a NaCl-rich hyposmotic saliva containing (mM): Na(+) 95 +/- 2; K(+) 38 +/- 1; Cl(-) 145 +/- 3. Saliva collected during serotonin stimulation had a similar composition. Bumetanide decreased secretion rates induced by dopamine and serotonin; secreted saliva had lower Na(+), K(+) and Cl(-) concentrations and osmolarity. Ouabain caused increased secretion rates on a serotonin background. Saliva secreted during dopamine but not serotonin stimulation in the presence of ouabain had lower K(+) and higher Na(+) and Cl(-) concentrations, and was isosmotic. We concluded: The Na(+)-K(+)-2Cl(-) cotransporter is of cardinal importance for electrolyte and fluid secretion. The Na(+)/K(+)-ATPase contributes to apical Na(+) outward transport and Na(+) and K(+) cycling across the basolateral membrane in acinar P cells. The salivary ducts modify the primary saliva by Na(+) reabsorption and K(+) secretion, whereby Na(+) reabsorption is energized by the basolateral Na(+)/K(+)-ATPase which imports also some of the K(+) needed for apical K(+) extrusion. PMID- 12769996 TI - Juvenile hormone accelerates ovarian development and does not affect age polyethism in the primitively eusocial wasp, Ropalidia marginata. AB - Juvenile hormone modulates post-imaginal reproductive division of labor in primitively eusocial species and promotes the production of queens (e.g., Polistes) while it modulates age polyethism and promotes the production of foragers in highly eusocial species (e.g., the honey bee). Ropalidia marginata is a primitively eusocial wasp that shows both post-imaginal regulation of reproductive division of labor as well as age polyethism. Hence, R.marginata is a particularly interesting model system to study the effect of juvenile hormone. We demonstrate here that a single, topical application of 100 micro g of juvenile hormone-III per female wasp accelerates ovarian development of wasps held in isolation. Similar application to wasps released back on to their natal nests has no effect on their rate of behavioral development as witnessed from the age of first performance of feed larva, build, bring pulp and bring food. We conclude therefore that in R.marginata, juvenile hormone has retained its function of modulating reproductive division of labor and has not acquired the function of modulating age polyethism. PMID- 12769997 TI - Effects of DIMBOA on detoxification enzymes of the aphid Rhopalosiphum padi (Homoptera: aphididae). AB - The presence of glutathione transferases and esterase activity was investigated in Rhopalosiphum padi and the effects of the cereal hydroxamic acid, 2,4 dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA) on these detoxification enzymes was studied. Activity of glutathione S-transferases and general esterases was determined for adult aphids feeding on a natural diet lacking DIMBOA and on an artificial DIMBOA-containing diet for 48 hours. In vivo, DIMBOA in the diet inhibited the activities of esterases by 50-75% at all concentrations tested (0.5 4 mM). The activity of glutathione transferase was inhibited to a lesser extent (30%) at the higher concentrations of DIMBOA. In vitro, DIMBOA generally inhibited the activity of esterases with an IC(50) of 33 micro M, and had a slight inhibitory effect on glutathione S-transferases. These effects of DIMBOA could make the aphids vulnerable to electrophilic agents and insecticides which may be metabolized via esterases and GSTs. In cereals, therefore, DIMBOA may act by interfering with esterase- or GST-mediated detoxification of xenobiotics by aphids. PMID- 12769998 TI - Pigment-dispersing factor sets the night state of the medulla bilateral neurons in the optic lobe of the cricket, Gryllus bimaculatus. AB - Pigment-dispersing factor (PDF) is an octadeca-neuropeptide widely distributed in the insect brain and suggested to be involved in the insect circadian systems. We have examined its effects on the neuronal activity of the brain efferents in the optic stalk including medulla bilateral neurons (MBNs) in the cricket, Gryllus bimaculatus. The MBNs are visually responding interneurons connecting the bilateral medulla, which show a clear day/night change in their light responsiveness that is greater during the night. Microinjection of PDF into the optic lobe induced a significant increase in the spontaneous activity of the brain efferents and the photo-responsiveness of the MBNs during the day, while little change was induced during the night. The enhancing effects began to occur about 20 min after the injection and another 10 min was necessary to reach the maximal level. The effects of PDF were dose-dependent. When 22 nl of anti-Gryllus PDF (1:200) IgG was injected into the medulla, the photo-responsiveness of the MBNs was suppressed in both the day and the night with greater magnitude during the night. No significant suppression was induced by injection of the same amount of IgG from normal rabbit serum. These results suggest that in the cricket optic lobe, PDF is released during the night and enhances MBNs' photo-responsiveness to set their night state. PMID- 12769999 TI - Temporal, spatial and induced expression of chitinase in the spruce budworm, Choristoneura fumiferana. AB - Temporal, spatial and induced expression of Choristoneura fumiferana chitinase (CfChitinase) was studied using immunohistochemistry and Western blots. CfChitinase was detected in the integument, the midgut peritrophic membrane, the cuticular lining of the trachea, the spiracle, and salivary glands. The enzyme was expressed as larvae were preparing to molt from one instar to the next. The spatial and temporal expression patterns are consistent with its function in degrading chitin during the molting process. The 20-hydroxyecdysone agonist, tebufenozide (RH5992), induced the expression of the CfChitinase gene in the early stage of the sixth-instar larvae and the enzyme was detected in the epidermis and molting fluid 24 h post treatment. PMID- 12770000 TI - Sequential deposition of yolk components during oogenesis in an insect, Aedes aegypti (Diptera: Culicidae). AB - Vitellogenesis in Aedes aegypti of uniform body size was followed at 27 degrees C in narrow time intervals throughout their first reproductive cycle by measuring the length, diameter, and volume of follicles and oocytes, the latter as an expression of the yolk mass (vitellus). Independent of all experimental conditions, a two-step process of elongation was recognized for both follicle length and yolk length, so that growth curves were consistently composed of two linear regressions with different slopes against time. Follicle lengths started to increase immediately after the blood meal, while oocytes took up to 6 h to show a measurable increase in yolk length. The first linear phase continued until 30 h, when yolk length reached 268+/-22 micro m. At this point, a transition occurred where the linearity shifted sharply for the next 6 h to 2-4-times higher slopes for both regressions. This second growth phase represented a 40% elongation of oocytes and follicles. Then, both curves leveled off at their final size, characteristic of mature ovaries: 462+/-10 micro m for oocytes, 489+/-11 micro m for follicles. These values remained constant until oviposition. The first linear growth phase was associated with an equicaloric and synchronous protein and lipid incorporation into the oocytes; levels of these substances reached their maximum by the end of this first phase and remained constant until oviposition. The second linear growth phase was characterized by rapid glycogen incorporation into oocytes from 20 to 100% of the maximum. Subsequently, the surface pattern of the exochorion became visible, marking the end of yolk incorporation. Since eggs are always laid on moist substrates, within 2-3 h of oviposition they double in volume and fresh weight, driven by more than tripling of their water content. When blood-fed females were exposed to five different temperatures between 17 and 37 degrees C, the distinction between the two linear growth phases persisted, but the slopes of the respective regressions, and therefore their durations, were affected. Eggs still matured at 37 degrees C but never hatched and at 12 degrees C only 18% hatched, whereas at all the intermittent temperatures hatching was 80-90%. Oogenesis appears to be limited to the range between 12 and about 32 degrees C. The effects of age, maternal body size and the source of the blood on vitellogenesis were also examined. These parameters affected the onset and/or extent of oogenesis in various ways. PMID- 12770001 TI - Effects of starvation and desiccation on energy metabolism in desert and mesic Drosophila. AB - Energy availability can limit the ability of organisms to survive under stressful conditions. In Drosophila, laboratory experiments have revealed that energy storage patterns differ between populations selected for desiccation and starvation. This suggests that flies may use different sources of energy when exposed to these stresses, but the actual substrates used have not been examined. We measured lipid, carbohydrate, and protein content in 16 Drosophila species from arid and mesic habitats. In five species, we measured the rate at which each substrate was metabolized under starvation or desiccation stress. Rates of lipid and protein metabolism were similar during starvation and desiccation, but carbohydrate metabolism was several-fold higher during desiccation. Thus, total energy consumption was lower in starved flies than desiccated ones. Cactophilic Drosophila did not have greater initial amounts of reserves than mesic species, but may have lower metabolic rates that contribute to stress resistance. PMID- 12770002 TI - The olfactory responses of the antenna and maxillary palp of the fleshfly, Neobellieria bullata (Diptera: Sarcophagidae), and their sensitivity to blockage of nitric oxide synthase. AB - The relative sensitivities of the olfactory receptors in the antenna and maxillary palp of the fleshfly, Neobellieria bullata, were assessed using simultaneous electroantennograms (EAGs) and electropalpograms (EPGs). In general, the antennae and maxillary palps were more sensitive to odors related to animals (blood extract and saturated carboxylic acid) than to odors that were plant derived (citral, hexenol, hexenal). In addition, the maxillary palps were relatively less sensitive to plant-derived odorants than the antennae, perhaps related to their anatomical position. Scanning electron microscopy was also used to assess the types of sensilla found on the two organs. In addition, NADPH diaphorase histochemistry was used in an attempt to localize the enzyme nitric oxide synthase (NOS) in the antenna and the maxillary palps. We found evidence of NADPH-diaphorase staining in both organs, with localized staining in the antennal cells and more general staining in the maxillary palps. When NOS was selectively blocked using the antagonist L-NAME, the amplitude of the EAGs and EPGs to odorants fell by 30-50%. In contrast, application of the inactive enantiomer, D NAME, did not change the amplitude of the EAGs or the EPGs. Our results indicate that NOS is involved in the function of olfactory receptor cells in the fleshfly. PMID- 12770003 TI - Oosorption in the stink bug, Plautia crossota stali: induction and vitellogenin dynamics. AB - Oosorption, resorption of developing oocytes in the ovary, in P. c. stali is characterized by changes in appearance of oocytes from opaque greyish green or orange to transparent, degeneration of yolk granules and disappearance of oocyte contents. Starvation and virginity were indicated to be factors that induce oosorption. SDS PAGE/Western blotting analysis using anti-vitellogenin antiserum detected two major and many minor bands in haemolymph samples. Egg extracts showed a more complicated set of positive bands in the same analysis. Yolk protein, vitellin, therefore, seemed to be formed after complicated processing of vitellogenin following its uptake by the oocytes. In starved, oosorption-induced females, vitellogenin concentration in the haemolymph was lower than that of fed females, and Western blotting failed to detect either oosorption-specific or ovary-specific peptide fragments in haemolymph samples collected from those females. These results suggest that once oosorption was induced vitellogenin/vitellin in oocytes was degraded rapidly and released into the haemolymph in the form of amino acids or small peptides too small to be recognized by the anti-vitellogenin antiserum. PMID- 12770004 TI - The role of hemolymph proline as a nitrogen sink during blood meal digestion by the mosquito Aedes aegypti. AB - Mosquitoes utilize the amino acids derived from blood meal protein to produce egg proteins. But the amino acids can also be used to produce egg lipid or can be oxidized for energy production. These latter two processes result in the release of nitrogen as toxic ammonia. Therefore, amino acids must be processed in such a way that amino acid nitrogen can be incorporated into non-toxic waste products. Proline is the predominant amino acid in the hemolymph of the adult female mosquito Aedes aegypti. After feeding on albumin meal, hemolymph proline levels increased five-fold over unfed levels, reached maximal levels in the first hours after feeding and remained high through oviposition. Hemolymph proline levels increased as the concentration of protein in the meal increased. When starved of sugar for 24 h prior to feeding on an albumin meal, hemolymph proline levels increased four-fold over the proline levels of non-starved mosquitoes. Proline levels after feeding on a protein deficient in essential amino acids, pike parvalbumin, increased to twice the levels of albumin fed mosquitoes. Based on these observations, we propose that mosquitoes utilize proline as a temporary nitrogen sink to store ammonia arising from deamination of blood meal amino acid. PMID- 12770005 TI - Chilling affects allatal cell proliferation via antennae and protocerebral neurons in the cockroach Diploptera punctata. AB - The corpora allata (CA) cells in a mated female of the cockroach Diploptera punctata undergo numerous mitotic divisions before an increase in juvenile hormone synthesis. A previous study demonstrated that this mitotic wave could be suppressed by exposure of the mated female to melting ice. Herein, we report that chilling suppresses CA mitosis via antennal perception. Cell proliferation suppressing stimuli from chilling were acquired in proportion to the length of time of exposure to the low temperature and the physical length of the antennae exposed to chilling. Sixty basal antennal annuli should remain exposed to chilling for at least 1.5 h in order to suppress mitotic divisions in CA. Mitotic divisions in corpus allatum are suppressed by stimuli from contralateral antenna, predominantly via pars intercerebralis neurons. Selective disconnection of pars intercerebralis neurons from CA, prior to chilling, restored the mitotic wave in CA. Cellular divisions did not occur in CA of chilled females if either pars lateralis neurons were severed or left intact. PMID- 12770007 TI - Lipogenesis in an adult parasitic wasp. AB - The goal of this study was to determine the extent of lipogenesis in the parasitoid Eupelmus vuilletti (Hymenoptera: Eupelmidae). Carbohydrate and lipid metabolism were followed in glucose-fed and starved females over 3 days. Fed females increased their glycogen level, while maintaining their lipid level. Starved females used most of their glycogen, while maintaining their lipid level too. Thus, females either use exclusively sugars to preserve their lipid reserves, or maintained a steady renewal of lipids through lipogenesis. The incorporation of radioactively marked glucose into lipids showed however that lipogenesis did not occur at a sufficient level to increase lipid reserves and to compensate for lipid use. This result has important implications for understanding nutrients allocation strategy in this species as the amount of lipids is almost totally fixed upon the emergence. From an evolutionary perspective, we call for detailed physiological studies of lipogenesis in a wide range of adult hymenopterans, as the absence of lipogenesis could be common to all of Aculatea. PMID- 12770006 TI - In vivo activation of pro-form Bombyx cysteine protease (BCP) in silkmoth eggs: localization of yolk proteins and BCP, and acidification of yolk granules. AB - The present study was designed to investigate the process of acidification of yolk granules during embryogenesis. In oocytes of mature Bombyx mori silkmoth, yolk proteins and a cysteine protease (pro-form BCP) were found in yolk granules. BCP was localized in small sized yolk granules (SYG, 3-6 microm in diameter) and yolk proteins in large sized granules (LYG, 6-11 microm in diameter), which might result in a spatial separation of protease and its substrates to avoid unnecessary hydrolysis. The granules were isolated on Percoll density gradient centrifugation. Although separation of LYG and SYG was incomplete, the granules sedimented in different fractions when using unfertilized egg extract, in which LYG was recovered from heavier fractions and BCP from lighter fractions. Acid phosphatase, as well as other lysosomal marker enzymes tested, was recovered from LYG-containing fractions. When extracts were prepared from developing eggs (day 3), some BCP-containing granules co-sedimented with LYG. The inactive pro-form BCP was activated in vivo, in parallel with yolk protein degradation, and as demonstrated previously in vitro under acidic conditions (). These results suggest that acidification occurs in yolk granules during embryogenesis. This was also confirmed using acridine orange fluorescent dye. In early development, most yolk granules were neutral, but became acidic during embryonic development. SYG were progressively recovered in heavier density fractions, displaying acidic interior. In this fraction, BCP-containing granules seem to be associated with larger granules (6-11 microm in size). In addition, SYG (BCP containing granules) were likely to be acidified earlier than LYG. Our results suggest that acidification initiates yolk degradation through activation of pro-form BCP. PMID- 12770008 TI - Venom of ectoparasitoid, Euplectrus sp near plathypenae (Hymenoptera: Eulophidae) regulates the physiological state of Pseudaletia separata (Lepidoptera: Noctuidae) host as a food resource. AB - Euplectrus sp. near plathypenae is an ectoparasitoid that can parasitize from 3rd to day 0-6th instar Pseudaletia separata. The developmental period of the parasitoid from the egg to the pupal stage is about 13 days. Parasitized hosts are developmentally arrested and never molt to the next stadium. The injection of venom fluid results in similar effects on P. separata larvae as does parasitization. The inhibitory effect of the venom on molting was dose dependent. Injection of 0.3 female equivalents of venom into day 0-5th host instar resulted in a similar developmental arrest as seen in parasitized hosts. The amount of total lipid in the hemolymph of the host increased as a function of the amount of venom injected, while the lipid content of the fat body was similar to lipid levels in the fat body of parasitized larvae. The amount of total protein in the hemolymph also increased when venom was injected, whereas the protein level of the fat body did not increase. The lipid concentration within the parasitoid larva was maintained at the same level throughout larval development, but increased before pupation. We conclude that the injected venom increased the hemolymph content of lipid and protein to support the growth and development of the ectoparasitoid larva. PMID- 12770009 TI - Isolation and characterization of three diapause-associated transcripts from the Colorado potato beetle, Leptinotarsa decemlineata. AB - Using suppressive subtractive hybridization, fragments of three diapause associated transcripts (DAT-1, 2 and 3) were isolated from the Colorado potato beetle, Leptinotarsa decemlineata. Full-length clones were developed for the transcripts. DAT-1 encodes a deduced protein 286 amino acids in length with limited identity to several proteins with leucine-rich domains. DAT-2 encodes a deduced protein 229 amino acids in length with 27% identity, 40% similarity to the desiccation stress protein from Tenebrio molitor. DAT-3 encodes a deduced protein 97 amino acids in length with identity to no known protein. DAT-1 and 2 have similar expression patterns as determined by northern blot analysis. Trace levels of these two transcripts are first detected in 3-day-old diapause programmed adults with a significant increase in expression on day 6. Expression of DAT-3 begins on day 12 in diapause-programmed adults and expression levels increase until the beetles enter diapause. Expression of DAT-1, 2 and 3 continues at least 60 days into diapause. PMID- 12770010 TI - Characterization of Na(+) currents in isolated dorsal unpaired median neurons of Locusta migratoria and effect of the alpha-like scorpion toxin BmK M1. AB - A primary cell culture was developed for efferent dorsal unpaired median (DUM) neurons of the locust. The isolated somata were able to generate Tetrodotoxin (TTX)-sensitive action potentials in vitro. The alpha-like scorpion toxin BmK M1, from the Asian scorpion Buthus martensi Karsch, prolonged the duration of the action potential up to 50 times. To investigate the mechanism of action of BmK M1, the TTX-sensitive voltage gated Na(+) currents were studied in detail using the whole cell patch clamp technique. BmK M1 slowed down and partially inhibited the inactivation of the TTX-sensitive Na(+) current in a dose dependent manner (EC50=326.8+/-34.5 nM). Voltage and time dependence of the Na(+) current were described in terms of the Hodgkin-Huxley model and compared in control conditions and in the presence of 500 nM BmK M1. The BmK M1 shifted steady state inactivation by 10.8 mV to less negative potentials. The steady state activation was shifted by 5.5 mV to more negative potentials, making the activation window larger. Moreover, BmK M1 increased the fast time constant of inactivation, leaving the activation time constant unchanged. In summary, BmK M1 primarily affected the inactivation parameters of the voltage gated Na(+) current in isolated locust DUM neurons. PMID- 12770011 TI - Possible function of two insect phospholipid-hydroperoxide glutathione peroxidases. AB - We compared the functional properties of two insect members of the phospholipid hydroperoxide glutathione peroxidases (PHGPx) family, VLP1, a major component of virus-like particles from the hymenopteran endoparasitoid Venturia canescens and its closest Drosophila relative, one of the putative PHGPx-proteins predicted from the Berkeley Drosophila genome sequence project. Recombinant Drosophila PHGPx shows enzymatic activity towards a number of PHGPx substrates, while the recombinant PHGPx-like domain of VLP1 lacks a functionally relevant cysteine and enzyme activity. A possible function of a non-enzymatic extracellular PHGPx-like protein is discussed. PMID- 12770012 TI - Midgut adaptation and digestive enzyme distribution in a phloem feeding insect, the pea aphid Acyrthosiphon pisum. AB - Transmission electron micrographs of the pea aphid midgut revealed that its anterior region has cells with an apical complex network of lamellae (apical lamellae) instead of the usual regularly-arranged microvilli. These apical lamellae are linked to one another by trabeculae. Modified perimicrovillar membranes (MPM) are associated with the lamellae and project into the lumen. Trabeculae and MPM become less conspicuous along the midgut. The most active A. pisum digestive enzymes are membrane-bound. An aminopeptidase (APN) is described elsewhere. An alpha-glucosidase (alpha-Glu) has a molecular mass of 72 kDa, pH optimum 6.0 and catalyzes in vitro transglycosylations in the presence of an excess of the substrate sucrose. There is a major cysteine proteinase activity (CP) on protein substrates that has a molecular mass of 40 kDa, pH optimum 5.5, is inhibited by E-64 and chymostatin and is activated by EDTA+cysteine. The enzyme is more active against carbobenzoxy-Phe-Arg-4-methylcoumarin-7-amide (ZFRMCA) than against ZRRMCA. These features identify the purified CP as a cathepsin-L-like cysteine proteinase. Most CP is found in the anterior midgut, whereas alpha-Glu and APN predominate in the posterior midgut. With the aid of antibodies, alpha-Glu and CP were immunolocalized in cell vesicles and MPM, whereas APN was localized in vesicles, apical lamellae and MPM. The data suggest that the anterior midgut is structurally reinforced to resist osmotic pressures and that the transglycosylating alpha-Glu, together with CP and APN are bound to MPM, thus being both distributed over a large surface and prevented from excretion with honeydew. alpha-Glu frees glucose from sucrose without increasing the osmolarity, and CP and APN may process toxins or other proteins occasionally present in phloem. PMID- 12770013 TI - Glucose and ecdysteroid increase apyrene sperm production in in vitro cultivation of spermatocysts of Bombyx mori. AB - Two types of sperm, nucleate eupyrene and anucleate apyrene, occur in the silkworm as in other lepidopteran species. Hormones and other substances have been assumed to play important roles in sperm dimorphism. We established an in vitro cultivation system for silkworm spermatocytes, and found that apyrene sperm are not produced when spermatocytes from larval testes are cultivated, though eupyrene spermatocytes develop normally into mature sperm. Based on the fact that ecdysteroid titers increase rapidly and peak 1 day after spinning, and that the amount of glycogen reaches its peak 1 day before the spinning stage, we studied the effects of adding glucose and/or 20-hydroxyecdysone to the culture medium. The experiments disclosed a significant additive effect of both substances on apyrene sperm production. PMID- 12770014 TI - Zinc is incorporated into cuticular "tools" after ecdysis: the time course of the zinc distribution in "tools" and whole bodies of an ant and a scorpion. AB - An understanding of the developmental course of specialized accumulations in the cuticular "tools" of arthropods will give clues to the chemical form, function and biology of these accumulations as well as to their evolutionary history. Specimens from individuals representing a range of developmental stages were examined using MeV - Ion microscopy. We found that zinc, manganese, calcium and chlorine began to accumulate in the mandibular teeth of the ant Tapinoma sessile after pre-ecdysial tanning, and the zinc mostly after eclosion; peak measured zinc concentrations reached 16% of dry mass. Accumulations in the pedipalp teeth, tarsal claws, cheliceral teeth and sting (aculeus) of the scorpion Vaejovis spinigeris also began after pre-ecdysial tanning and more than 48 h after ecdysis of the second instars. Zinc may be deposited in the fully formed cuticle through a network of nanometer scale canals that we observed only in the metal bearing cuticle of both the ants and scorpions. In addition to the elemental analyses of cuticular "tools", quantitative distribution maps for whole ants were obtained. The zinc content of the mandibular teeth was a small fraction of, and independent of, the total body content of zinc. We did not find specialized storage sites that were depleted when zinc was incorporated into the mandibular teeth. The similarities in the time course of zinc, manganese and calcium deposition in the cuticular "tools" of the ant (a hexapod arthropod) and those of the scorpion (a chelicerate arthropod) contribute to the evidence suggesting that heavy metal halogen fortification evolved before these groups diverged. PMID- 12770015 TI - Rapid responses to high temperature and desiccation but not to low temperature in the freeze tolerant sub-Antarctic caterpillar Pringleophaga marioni (Lepidoptera, Tineidae). AB - A broad definition of rapid cold hardening (RCH) is that it is the process whereby insects increase their survival of a sub-zero temperature after a brief (h) pre-exposure to a less severe low temperature. The effects of various pre treatments on survival of two h at -7.9 degrees C were investigated in the freeze tolerant sub-Antarctic caterpillar Pringleophaga marioni (Lepidoptera: Tineidae), the first time RCH has been investigated in a freeze tolerant arthropod. All caterpillars froze when exposed to -7.9 degrees C, and none of the low temperature pre-treatments (-5, 0, 5 and 15 degrees C, as well as -5 degrees C and 0 degrees C with a delay before freezing) nor slow cooling (0.1 degrees C/min) elicited any improvement in survival of -7.9 degrees C as compared to controls. However, high temperature treatments (25, 30 and 35 degrees C), desiccation and acclimation for 5 days at 0 degrees C did result in significant increases in survival of the test temperature, possibly as a result of heat shock protein production. Haemolymph osmolality was elevated only by the 35 degrees C pre-treatment. It is suggested that the unpredictable environment of Marion Island means that P. marioni must always be physiologically prepared to survive cold snaps, and that this year-round cold hardiness therefore supersedes a rapid cold hardening response. PMID- 12770016 TI - Developmental and diel changes of adipokinetic hormone in CNS and haemolymph of the flightless wing-polymorphic bug, Pyrrhocoris apterus (L.). AB - We have used an enzyme-linked immunoassay (ELISA) for determination of the AKH content in CNS (brain + corpora cardiaca + corpora allata) and haemolymph of adult macropterous and brachypterous females of the bug Pyrrhocoris apterus. The tests revealed that the AKH content fluctuates between 1 and almost 4 pmol/CNS during the first 14 days of adult life and significantly increased in order diapausing brachypters/=16 degrees C). In contrast to a general rule, the PE is less unsaturated than PC. In both TM and FB, proportions of unsaturated/unsaturated molecular species are consistently high in PC (56.3-67.5% in TM, 59.2-66.6% in FB), while they are consistently low in PE (19.1-26.7% in TM, 12.1-15.1% in FB). An adaptive significance of changes in the phospholipid composition for the low temperature and/or dehydration stress is discussed in relation to known physical properties of phospholipids. PMID- 12770025 TI - Response of storage protein levels to variation in dietary protein levels. AB - Storage proteins have been found to play a major role in insect metamorphosis and egg production and are accumulated during the actively feeding larval stage. Yet few studies have focused on how nutrition affects storage protein levels. Three storage proteins were identified in male and female Heliothis virescens pupae, one arylphorin and two putative high-methionine hexamers. Storage proteins were quantified in early pupae and in pharate adults. Storage protein levels peaked in 48-h pupae and were more abundant in females across all stages. Both male and female pharate adults retained a portion of total storage protein levels and females retained greater levels overall. In females, post-eclosion protein reserves will likely be used toward egg manufacturing, while the role of protein reserves in males remains speculative. In our previous study of H. virescens larvae, we found that protein-derived growth in females progressively increased as dietary protein levels increased. Our present data show that levels of storage protein also increased progressively along with dietary protein levels. This suggests that females allocated protein, in excess of adult tissue formation needs, toward storage protein. Our study is the first to demonstrate how responsive storage protein levels can be in face of varying levels of dietary protein. PMID- 12770026 TI - Entering diapause is a prerequisite for successful cold-acclimation in adult Graphosoma lineatum (Heteroptera: Pentatomidae). AB - In diapause adults of Graphosoma lineatum overwintering in a field-cage, high chill-tolerance (CT) developed gradually, within 5 months from August to December. In laboratory-acclimation experiments, the diapause state appeared to be an essential pre-condition for successful cold-acclimation and overwintering. First, diapause prevented elevation of the median supercooling point (SCP) by about 5.5 degrees C that accompanies the onset of reproductive activity in non diapause specimens. Second, diapause allowed subsequent physiological changes resulting in cold-acclimation during a gradual (18-day) decrease of temperature from 25 to 0 degrees C. No, or very modest, cold-acclimation was observed in non diapause specimens. Decrease of temperature led to a rapid loss of ca. 1/3 of the body water in both non-diapause and diapause specimens. Approximately 0.1 M of trehalose accumulated in tissues of diapause specimens only, and haemolymph osmolality rose from 347 mOsm (at 25 degrees C) to 444 mOsm after an 18-day cold acclimation and to 764 mOsm during further storage at 0 degrees C for 100 days. Upon transfer of cold-acclimated diapause specimens back to 25 degrees C for one week (de-acclimation), the high CT was lost, the SCP elevated by about 2.5-3 degrees C, and the levels of trehalose, water content and haemolymph osmolality returned to pre-acclimation or non-diapause levels. PMID- 12770027 TI - Larvae of an endoparasitoid, Cotesia kariyai (Hymenoptera: Braconidae), feed on the host fat body directly in the second stadium with the help of teratocytes. AB - Larvae of a gregarious endoparasitoid, Cotesia kariyai (Watanabe), grew rapidly during the second stadium in the host. The fat body of a Pseudaletia host parasitized by C. kariyai was completely consumed by 10 d, just before larval emergence. It seemed hard to explain the growth of the second instar parasitoids and the rapid consumption of the fat body only by ingestion of hemolymph converted from the fat body or other organs of the host. Paraffin sections of the parasitized host revealed that many teratocytes were attached to the surface of the fat body in many sites and destroyed the fat body tissue locally. Zymography of proteins released from the teratocytes revealed that the teratocytes 4 to 9 days after parasitization showed collagenase activity (as a gelatinase). Further, 1st instar parasitoids which were transplanted together with teratocytes into unparasitized hosts preconditioned with C. kariyai polydnavirus (CkPDV) plus venom, grew normally to the 2nd stadium. Abnormal growth of parasitoid larvae was observed when parasitoid larvae were transplanted without teratocytes. These results suggest that the teratocytes attach to the outer sheath of the fat body, secrete an enzyme that makes a hole in the matrix of the fat body, thus allowing the second instar parasitoid to ingest the content of the fat body. PMID- 12770028 TI - The ectoparasitic wasp Nasonia vitripennis (Walker) (Hymenoptera: Pteromalidae) differentially affects cells mediating the immune response of its flesh fly host, Sarcophaga bullata Parker (Diptera: Sarcophagidae). AB - In this study, we examined cellular immune responses in the flesh fly, Sarcophaga bullata, when parasitized by the ectoparasitoid Nasonia vitripennis. In unparasitized, young pharate adults and third instar, wandering larvae of S. bullata, four main hemocyte types were identified by light microscopy: plasmatocytes, granular cells, oenocytoids, and pro-hemocytes. Parasitism of young pharate adults had a differential effect on host hemocytes; oenocytoids and pro-hemocytes appeared to be unaltered by parasitism, whereas adhesion and spreading behavior were completely inhibited in plasmatocytes and granular cells by 60 min after oviposition. The suppression of spreading behavior in granular cells lasted the duration of parasitism. Plasmatocytes were found to decline significantly during the first hour after parasitism and this drop was attributed to cell death. Melanization and clotting of host hemolymph did not occur in parasitized flies, or the onset of both events was retarded by several hours in comparison to unparasitized pharate adults. Hemocytes from envenomated flies were altered in nearly identical fashion to that observed for natural parasitism; the total number of circulating hemocytes declined sharply by 60 min post envenomation, the number of plasmatocytes declined but not granular cells, and the ability of plasmatocytes and granular cells to spread when cultured in vitro was abolished within 1 h. As with parasitized hosts, the decrease in plasmatocytes was due to cell death, and inhibition of spreading lasted until the host died. Isolated crude venom also blocked adhesion and spreading of these hemocyte types in vitro. Thus, it appears that maternally derived venom disrupts host immune responses almost immediately following oviposition and the inhibition is permanent. The possibility that this ectoparasite disables host defenses to afford protection to feeding larvae and adult females is discussed. PMID- 12770029 TI - The dark-color inducing neuropeptide, [His(7)]-corazonin, causes a shift in morphometic characteristics towards the gregarious phase in isolated-reared (solitarious) Locusta migratoria. AB - The neurohormone, [His(7)]-corazonin is known to induce dark color in the cuticle and epidermis of Locusta migratoria. In the present study, we examined the effects of this hormone on development and morphometrics in two strains, albino and normal, of this locust under isolated conditions. Injection of [His(7)] corazonin induced dark color in both strains. In either strain, [His(7)] corazonin injected at the second and third instars did not affect duration of nymphal development or the number of nymphal instars. The shape of the pronotum was more convex in isolated-reared animals than in crowd-reared ones, and injection of [His(7)]-corazonin caused isolated-reared animals to develop a less convex pronotum in the normal strain injected at a high dose (1 nmolx2) but not in the albino strain injected at a low dose (50 pmolx2). [His(7)]-corazonin injected into isolated-reared nymphs caused a shift in classical morphometric ratios (F/C and E/F; F=length of the hind femur, C=maximum width of the head, E=length of the fore wings) towards values typical for crowd-reared (gregarious) individuals of the two strains. This study demonstrated for the first time that [His(7)]-corazonin affected morphometric characteristics in L. migratoria. PMID- 12770030 TI - Brain tyramine and reproductive states of workers in honeybees. AB - To explore the role of tyramine in the transformation of reproductive states of honeybee workers, brain levels of tyramine and N-acetyltyramine were measured in both normal and queenless workers. Queenless workers had higher tyramine levels and lower N-acetyltyramine levels than normal workers did. Intermediate reproductive workers that were transferred into a normal colony from a queenless colony had intermediate levels of tyramine and N-acetyltyramine. Elevation of tyramine in the queenless workers occurred at an earlier adult stage than elevation of dopamine. Tyramine levels in intermediate reproductive workers returned to the levels seen in normal workers, but dopamine levels in intermediate reproductive workers remained elevated at the same level as in queenless workers. Thus, brain tyramine may be regulated by the colony condition with or without a queen. Injection of an amine uptake inhibitor, reserpine, depleted tyramine and elevated N-acetyltyramine. Distributions of tyramine and dopamine within the brain were distinctively different, whereas distributions of N-acetyltyramine and N-acetyldopamine were similar, suggesting that each functional amine is stored in specific neurosecretory cells and released to the relevant receptor sites but that metabolism into each N-acetylmetabolite is determined by diffusion. PMID- 12770031 TI - Variation in the number of sperm transferred during mating among males of the Colorado potato beetle (Coleoptera: Chrysomelidae). AB - We suggest an explanation for the bimodal distribution in the number of sperm transferred into the female during mating by males of the Colorado potato beetle (Leptinotarsa decemlineata). While in most matings females receive 20,000 sperm, in others they receive less than 1000. A variety of potential proximate causes for this bimodal distribution including experimental procedures and several physiological and behavioral causes were examined: male body size, female body size, male mating history, socio-sexual environments, duration and number of mounts, and duration and number of copulations. Surprisingly, none of these explained the observed distribution. However, we did find that the total time spent in copulation was significantly less variable among males that transferred more than 20,000 sperm than among males that passed less than 1000 sperm. In addition, males transferring large amounts of sperm resulted in proportionally more sperm inside the female's spermatheca within a short period of time than males passing less than 1000 sperm. The lack of pattern in male sperm ejaculate suggests definite differences in sperm delivery tactics and may be related to the female condition rather than the male. PMID- 12770032 TI - Prey-mediated effects of the protease inhibitor aprotinin on the predatory carabid beetle Nebria brevicollis. AB - To investigate the potential non-target impacts of transgenic pest-resistant plants, prey-mediated impacts of a protease inhibitor (PI) on the predatory carabid, Nebria brevicollis, were investigated. The PI used was aprotinin, a serine PI of mammalian origin with insecticidal properties when incorporated in artificial diet or expressed in transgenic plants. Field-collected N. brevicollis adults, kept at 23 degrees C, 16:8 L:D, were fed, over their pre-aestivation activity period of 24 days, with Helicoverpa armigera larvae reared on an artificial diet containing 0.5% (w:w, fresh mass) aprotinin. These larvae contained 22.62 &mgr;g aprotinin/g insect. Control prey was reared on diet without aprotinin. Beetle survival and body mass were unaffected by prey type. Beetles consuming PI-fed prey lost significantly more mass than the control beetles during two periods of mass loss, but gained significantly more mass during the final period of mass gain. This was not due to differences in amounts of prey supplied or consumed. The final mass gain coincided with increased consumption of PI-prey. Female beetles were significantly heavier than males, but we found no consistent gender-based differences in response to PI-prey. At the end of the experiment, body mass of all beetles was similar to field-collected ones (approximately 55 mg). All experimental beetles had significantly lower activities of digestive cysteine proteases and the serine proteases chymotrypsin and trypsin than field-collected ones. Beetles consuming PI-fed prey had significantly lower levels of trypsin and higher levels of chymotrypsin and elastase than the control beetles. PMID- 12770033 TI - A quantitative study of adipokinetic hormone of the firebug, Pyrrhocoris apterus. AB - The development of an enzyme-linked immunoassay (ELISA) for the adipokinetic neuropeptide hormone, Pya-AKH, from the firebug Pyrrhocoris apterus L. is described. The ELISA measures as little as 20 fmol of Pya-AKH. Tested against a range of synthetic peptides, the assay has a high sensitivity for peptides containing the C-terminal motif FTPNWamide. The amounts of Pya-AKH in the brain, corpora cardiaca, suboesophageal ganglia, and fused thoracic and abdominal ganglionic mass are very small, with only the corpora cardiaca containing appreciable levels of the hormone (ca. 4 pmol per bug). Preliminary estimates of the persistence of the hormone in the haemolymph are consistent with values determined for AKHs in other insects, and suggest that Pya-AKH has a rapid turnover with a half-life of ca. 18 min. Measurements of circulating titres of AKH in Pyrrhocoris are only possible in the ELISA described here by using pooled samples of haemolymph, and after preliminary clean-up of the haemolymph samples. The titre of Pya-AKH in resting reproductive female Pyrrhocoris is ca. 1 fmol/&mgr;l. PMID- 12770034 TI - Plant odour processing in the antennal lobe of male and female grapevine moths, Lobesia botrana (Lepidoptera: Tortricidae). AB - Moths of Lobesia botrana (Lepidoptera: Tortricidae) are confronted with different volatiles emitted from the host plant during the different seasons. To test the hypothesis of plasticity of central plant odour processing in moths of different generations in the future, we first investigated the responses of antennal lobe (AL) interneurons of laboratory-reared virgin and mated males and females. We used intracellular recording and staining techniques while stimulating the antenna with a range of host and non-host plant odours. The AL structure of L. botrana is similar to that found in other Lepidoptera species studied. The most frequent physiological responses for all types of moths were obtained with (E)-2 hexenal, and with thujyl alcohol and beta-thujone, components of tansy, a behaviourally attractive non-host plant. Some broadly responding neurons were capable of distinguishing between different compounds through different response patterns (excitation/inhibition) and/or different dose-response characteristics. Response characteristics (response spectra, threshold and specificity) of neurons were similar, independent of sex or mating status of the moths. Significant differences between the groups were, however, found in the proportion of responding neurons for a few tested components. PMID- 12770035 TI - Juvenile hormone and circadian locomotor activity in the honey bee Apis mellifera. AB - Age-related division of labor in honeybees is associated with plasticity in circadian rhythms. Young nest bees care for brood around the clock with no circadian rhythms while older foragers have strong circadian rhythms that are used for sun compass navigation and for timing visits to flowers. Since juvenile hormone (JH) is involved in the coordination of physiological and behavioral processes underlying age-related division of labor in honey bees, we tested the hypothesis that JH influences the ontogeny of circadian rhythms and other clock parameters in young worker bees. Treatments with the JH analog methoprene or allatectomy did not influence the onset of rhythmicity, overall locomotor activity, or the free-running period of rhythmic locomotor behavior. There were, however, significant differences in the onset of rhythmicity, overall locomotor activity, and longevity between bees from different source colonies, suggesting that there is significant genetic variation for these traits. Our results suggest that JH does not coordinate all aspects of division of labor in bees and that coordination of task performance with circadian rhythms is probably mediated by other regulatory systems. PMID- 12770036 TI - Effect of freeze temperature on ice formation and long-term survival of the woolly bear caterpillar (Pyrrharctia isabella). AB - Tissue ice content and post-freeze survival were documented for caterpillars of the arctiid moth Pyrrharctia isabella. Tissue ice content was inversely dependent on freeze temperature (-3 degrees C=24.4%, -6 degrees C=40.2%, -10 degrees C=48.7%) but values were substantially less than expected given hemolymph osmolality. Accumulation of glycerol (200-300 mM) in the hemolymph helped to colligatively reduce the amount of freezable water. Caterpillars engaged in locomotion within minutes after thawing but mortality occurred over the ensuing weeks, with the highest level (52.2%) occurring in the -10 degrees C fast thaw group. Pupation rates ranged between 45.7 to 52.4% of caterpillars in a test group. Adult emergence exceeded 60% of the pupae in the -3 and -6 degrees C test groups. Hence, P. isabella caterpillars survived ecologically relevant freezes and continued their life cycles to adulthood. PMID- 12770037 TI - Improvement of signal-to-noise ratio in electroantennogram responses using multiple insect antennae. AB - Using an array of insect antennae connected in series or in parallel, electroantennogram (EAG) responses and noise levels were investigated in an attempt to improve signal-to-noise (S/N) ratio and sensitivity. Both the EAG response amplitude and noise level increased when the antennae of male Helicoverpa zea (Lepidoptera: Noctuidae) were connected in series. Due to lower relative increase in noise level than EAG amplitude as the number of antennae increased, the S/N ratio was also significantly improved by the serial connection. As a result the sensitivity of EAG was improved by the serial connection, which showed ca. ten-fold improvement in the threshold detection levels compared with a single antenna when four antennae were connected in series. In contrast to the serial connection, there were no differences in EAG amplitudes and overall noise levels when different numbers of antennae were connected in parallel. When only large-amplitude noise was taken into account, however, the S/N ratio was somewhat improved by the parallel connection. The frequency of overall noise remained at the same level both in the serial and in the parallel connection. However, the frequency of the large-amplitude noise increased in serial connection but decreased in parallel connection. The present study clearly indicates that both the sensitivity and S/N ratio of the EAG biosensor could be significantly improved by using the multiple antennal connections. PMID- 12770038 TI - Ion physiology of vitellogenic follicles. AB - The ion physiology of vitellogenic follicles from a lepidopteran (Hyalophora cecropia) and a hemipteran (Rhodnius prolixus) are compared. Similarities that can be expected to occur in vitellogenic follicles of many other insects include: (1) gap junctions, which unite the cells of a follicle into an integrated electrical system, (2) transmembrane K(+) and H(+) gradients that account for over 60% of follicular membrane potentials, (3) absence of a Cl(-) potential, (but the opening of channels to this anion when vitellogenesis terminates in H. cecropia), (4) an electrogenic proton pump that supplements follicular membrane potentials, (5) Ca(2+) action potentials evoked by injecting depolarizing currents into oocytes, and (6) the use of osmotic pressure to control epithelial patency. Differences include: a Na(+)/K(+)-ATPase that accounts for about 20% of the follicular resting potential in R. prolixus but is absent from H. cecropia, and an intrafollicular Ca(2+) current that moves from oocyte to nurse cells through cytoplasmic bridges in H. cecropia. Evidence is also summarized for two promising mechanisms that require further substantiation: (1) transmission via gap junctions of a follicle cell product that promotes endocytosis in the oocyte; and (2) transport of the proton pump back and forth between cell surface and endosomes as the membrane that carries it recycles through successive rounds of vitellogenin uptake. PMID- 12770039 TI - Sodium regulation during dehydration of a herbivorous and a carnivorous beetle from African dry savannah. AB - The sodium regulation of carnivorous carabid beetles of the genus Cypholoba and herbivorous tenebrionid beetles of the species Phrynocolus petrosus from dry savannah in East Africa was investigated while the beetles went through dehydration in the laboratory. In both species the water loss took place mainly at the expense of the extracellular fluid, and in both species the loss of extracellular water was accompanied by a loss of extracellular sodium. In the carabid beetles the sodium removed from the extracellular fluid was excreted from the body, while in the tenebrionids sodium was kept within the body. It is proposed that the different manners in which the two species handle their sodium reflect differences in their access to dietary water and sodium. PMID- 12770040 TI - Pupal commitment and its hormonal control in wing imaginal discs. AB - The timing of pupal commitment of the forewing imaginal discs of the silkworm, Bombyx mori, was determined by a transplantation assay using fourth instar larvae. The wing discs were not pupally committed at the time of ecdysis to the fifth instar. Pupal commitment began shortly after the ecdysis and was completed in 14 h. When the discs of newly molted larvae (0-h discs) were cultured in medium containing no hormone, they were pupally committed in 26 h. In vitro exposure of 0-h discs to 20-hydroxyecdysone accelerated the progression of pupal commitment. Methoprene, a juvenile hormone analog (JHA), did not suppress the change in commitment in vitro at physiological concentrations. Thus the wing discs at the time of the molt have lost their sensitivity to JH, and 20E is not a prerequisite for completion of pupal commitment. These results suggest that the change in commitment in the forewing discs may begin before the last larval molt. PMID- 12770041 TI - Semiochemicals from ovaries of gravid females attract ovipositing female houseflies, Musca domestica. AB - Chemical signals originating from the ovaries of gravid females of Musca domestica (Diptera: Cyclorrhapha: Muscidae) attract ovipositing females to common egg-laying sites. Behavioral experiments indicated that females preferred to oviposit in fermented wheat bran containing ovaries from reproductively mature houseflies. Females preferred to oviposit in fermented wheat bran than wet wheat bran. This effect was additive with the attraction to housefly ovaries. Solvent extracts from housefly ovaries were attractive to gravid females. Extracts obtained with hexane were most attractive to gravid females for egg laying, and extracts obtained with ethyl acetate attracted more egg laying than extracts obtained by dichloromethane. Gas chromatography (GC) and gas chromatography coupled with mass spectrometry (GC-MS) analysis showed that tricosane and (Z)-9 tricosene were the main components of the hexane extracts. Both tricosane and (Z) 9-tricosene were shown to elicit dose-dependent aggregation of gravid females in oviposition bioassays, but high doses of either chemical were not attractive. PMID- 12770042 TI - Mode of action of an insect neuropeptide leucopyrokinin (LPK) on pupariation in fleshfly (Sarcophaga bullata) larvae (Diptera: Sarcophagidae). AB - An insect neuropeptide leucopyrokinin (LPK) (pQTSFTPRLamide) accelerates pupariation in wandering larvae of the fleshfly Sarcophaga bullata. The period of sensitivity to the action of LPK begins approximately 4 h before pupariation. Within this period the degree of acceleration of contraction into the shape of a puparium is practically independent of the age at which the larvae are injected, while acceleration of tanning is distinctly more age dependent. From ligation experiments we conclude that intact central innervation is essential for the action of LPK on puparial contraction, whereas central neurones take no part in mediation of LPK action on tanning of the cuticle. An analysis of tensiometric recordings of muscular activity revealed that the actual time of LPK accelerated puparial contraction coincides with the beginning of the immobilisation/retraction phase. LPK accelerates the switch from wandering behaviour to immobilisation/retraction behaviour but has no effect on the onset and duration of motor patterns that normally underlie puparial contraction in controls. The morphology of an accelerated puparium is normal but its formation is temporally dissociated from normal 'contraction patterns' that are performed a long time after the puparium has contracted. It means that neuromuscular activity of larvae accelerated by LPK does not cease upon formation of the white puparium, but continues until the whole motor programme of pupariation behaviour is completed. Apparently the peptide acts on the integument by stimulating it to contract and shrink, and no specific patterns of muscular contractions are needed to properly shape the puparium. This finding sheds a new light on our understanding of the mechanism of puparium formation. PMID- 12770043 TI - Drought acclimation and lipid composition in Folsomia candida: implications for cold shock, heat shock and acute desiccation stress. AB - Many of the physiological adaptations evolved in terrestrial invertebrates to resist desiccation have also been shown to enhance the survival of low temperatures. In this study we have examined temporal changes in the physiology of the collembolan Folsomia candida during acclimation to mild desiccation stress (98.2% RH), and how physiological changes correlate with resistance to subsequent cold shock, heat shock and acute desiccation stress. Drought-acclimation increased the resistance to cold and acute drought but reduced the resistance to heat shock. The composition of membrane phospholipid fatty acids (PLFA) changed during acclimation resulting in a higher degree of unsaturation by the end of the 192-h acclimation period. This resembles typical membrane alterations seen in ectothermic animals exposed to cold. Only small changes were seen in the neutral lipid fraction. The temporal changes in cold resistance and drought resistance correlated well with changes in PLFA composition and accumulation of sugars and polyols ('cryoprotectives'). It is proposed that the drought-induced PLFA desaturation, combined with the membrane protecting accumulation of cryoprotectives, are important physiological adaptations providing tolerance to both desiccation and cold. PMID- 12770044 TI - Pea aphid clonal resistance to the endophagous parasitoid Aphidius ervi. AB - The physiological mechanism of resistance to the endophagous braconid Aphidius ervi Haliday (Hymenoptera, Braconidae) by a pink clone (PC) of Acyrthosiphon pisum (Harris) (Homoptera, Aphididae) has been investigated. Comparative data on parasitoid development and associated host biochemical changes in the resistant PC aphids and in a susceptible green clone (GC) of A. pisum are reported. When the PC aphids were attacked as early 4th instars, the developing parasitoid larvae showed a strongly reduced increase in size, compared to those synchronously developing in GC aphids, and were unable to produce a regular mummy. In contrast, parasitism of 2nd instar PC aphids, allowed completion of parasitoid development, but adults had a prolonged developmental time, due to a longer duration of parasitoid's final (3rd) instar. In all cases, teratocytes, cells deriving from the A. ervi serosal membrane, and the proteins abundantly synthesised by them, were never found in the haemolymph of parasitised PC aphids. Host castration, as demonstrated by total protein incorporation into reproductive tissues, was total in the majority of early (2nd instar) parasitised host aphids, while it was limited when later instars (4th) of PC aphids were parasitised. This is partly due to the absence of the cytolytic activity of teratocytes on host embryos, which, through their persistence, may compete for nutritional resources with the developing parasitoid larvae. In parasitised PC aphids, this competitive effect is further aggravated for the parasitoid by the absence of the regulated amino acid titre increase in the host haemolymph, which is regularly observed in GC aphids. Failure of teratocyte development in the PC clone of the pea aphid is, then, the major functional constraint accounting for the reduction/inhibition of A. ervi larval growth. The reported results allow to assess in vivo the role of teratocytes in the host physiological redirection and nutritional exploitation by the parasitoid, and to integrate and validate the proposed physiological model of host-parasitoid interactions in the system A. pisum-A.ervi. PMID- 12770045 TI - Effects of [His(7)]-corazonin on the phase state of isolated-reared (solitarious) desert locusts, Schistocerca gregaria. AB - [His(7)]-corazonin is a neuropeptide produced in the pars lateralis of the brain. It is stored in the corpora cardiaca and probably released from there. The only well-documented effect in locusts is increased melanization of the cuticle. We investigated whether this hormone might also be causally related to changes in behavior and morphometrics that, like melanization, occur during crowding-induced gregarization. Solitary fourth-instar nymphs of Schistocerca gregaria (Forsk.) were injected thrice with 1 nmol [His(7)]-corazonin. After molting to the 5th stadium their behavioral phase state was measured in an arena assay and analyzed using multiple logistic regression analysis. The hormone was found not to induce behavioral phase changes. Upon reaching adulthood, morphometrics shifts occurred towards values typical for crowd-reared and regregarized animals. Our results thus indicate that [His(7)]-corazonin is not involved in behavioral gregarization but may participate in morphometrical phase change. PMID- 12770046 TI - A ribosome-free extract from cultured cells improves recovery of polysomes from the mosquito fat body: analysis of vitellogenin and ribosomal protein rpL34 gene expression. AB - We have examined the association of ribosomal protein rpL34 mRNA with polysomes in Aedes albopictus C7-10 cells in culture using a simple, two-step sucrose gradient. In growing cells, 40-50% of the ribosomes were engaged on polysomes. This proportion could be increased to 80% when metabolism was stimulated by refeeding the cells with fresh medium. Conversely, ribosomes shifted off polysomes when cells were starved with phosphate-buffered saline or cell lysates were treated with puromycin. When similar approaches were used with fat body from blood-fed female Aedes aegypti mosquitoes, we were unable to obtain the polysome fraction that contained vitellogenin mRNA, which is abundantly translated after a blood meal. Addition of post-mitochondrial supernatant from fat body to polysomes from cultured cells shifted the polysome profile towards smaller polysomes and monosomes, in a dose-dependent fashion. Disruption of fat body tissue in a post ribosomal supernatant from refed cells improved the recovery of polysomes, demonstrating both the engagement of vitellogenin mRNA on polysomes and the mobilization of rpL34 from messenger-ribonuceloprotein particles onto polysomes in blood-fed mosquitoes. These observations suggested that ribonucleases remain active when polysomes are prepared from mosquito fat body, and that cell culture supernatant contains a ribonuclease inhibitor. PMID- 12770047 TI - Larvae of the ectoparasitic wasp, Eulophus pennicornis, release factors which adversely affect haemocytes of their host, Lacanobia oleracea. AB - When larvae of the ectoparasitic wasp Eulophus pennicornis were incubated for 4 h on balls of cotton wool soaked in tissue culture medium (TC-100), they released a variety of factors. Subsequent incubation of these larval wasp secretions with monolayers of haemocytes from their host, Lacanobia oleracea, demonstrated that they adversely affect haemocyte morphology, behaviour and viability. For instance, when monolayers of haemocytes were incubated for 18 h in TC-100, approximately 73% of the cells present, attached firmly to and spread over the tissue culture surface by extending pseudopods. By contrast, when incubated in TC 100 containing larval wasp secretions, only about 27% of the haemocytes present remained attached to the tissue culture surface after washing. The majority of these had a rounded configuration and neither spread nor extended pseudopods. Furthermore, viability assays indicated that approximately 36% of the attached haemocytes were dead, as opposed to 11-12% in the controls. The E. pennicornis secretions also significantly reduced the ability of L. oleracea haemocytes to move across the surface of the slide and form clumps (p50 &mgr;g. Parasitoids which failed to emerge eventually succumbed as unecydsed pharate third instar larvae in the hemocoel of the host. Effects of host methoprene treatment on parasitoid metamorphosis were also assessed, and metamorphic disruption occurred at much lower dosages compared with doses necessary to suppress parasitoid emergence behavior. The inhibitory effect of methoprene on parasitoid emergence behavior appears to be mediated by effects of this hormone on the synthesis or release of ecdysis-triggering hormone (ETH) in the parasitoid, the proximate endocrine cue which triggers ecdysis behavior in free-living insects. ETH accumulated in the epitracheal Inka cells of parasitoids developing in methoprene-treated hosts, suggestive of a lack of hormone release. Thus, the hormonal modulation of parasitoid emergence behavior appears to be complex, involving a suite of hormones including JH, ecdysteroid, and peptide hormones. PMID- 12770068 TI - Allatotropic activity in the brain of female Phormia regina (Diptera: Calliphoridae). AB - In Phormia regina, the rate of juvenile hormone (JH) synthesis rises rapidly after the ingestion of an adequate protein meal. In a previous publication we have localized the neurons containing Manduca sexta allatotropin (Mas-AT)-like substances in the brain of P. regina and demonstrated the allatotropic effect of synthetic Mas-AT in sugar-fed flies in vitro. In this current study, we examined the possible role of the brain of P. regina after the fly received a protein meal. In vitro studies showed that the brain releases, at 8 h after a protein meal, a factor(s) with a strong allatotropic effect. This factor(s) stimulates the corpus allatum (CA) to produce 6.9 times more juvenile hormones (JHs) than the control CA. Time course studies showed that the release of this allatotropic factor(s) is temporally controlled. Only the brains collected from flies at 6 and 8 h after the onset of a liver meal release allatotropic factor(s). Injection of anti-Mas-AT antiserum partially suppressed the fly follicle development in vivo. Presence of anti-Mas-AT antiserum decreased the allatotropic effect of the brain released allatotropic factor(s) in vitro. In addition to a Mas-AT-like substance, it is possible that the brains of liver-fed P. regina females may synthesize other allatotropic factors that are chemically unrelated or partially related to Mas-AT, which cannot be recognized/neutralized by our anti-Mas-AT antiserum. PMID- 12770069 TI - Possible involvement of ecdysteroids in embryonic diapause of Locusta migratoria. AB - In the Ibaraki population (Japan) of Locusta migratoria, adult locusts produce diapause eggs under short-day (SD) conditions and non-diapause eggs under long day (LD) conditions. The identity and titre of ecdysteroids in the ovaries and eggs from LD and SD adult females were investigated by RIA/HPLC. Maternal ecdysteroids accumulated in the developing ovaries represented about 90% polar conjugates, 5% free ecdysteroids and 5% non-hydrolyzable metabolites. Before oviposition the quantity of ecdysteroids reached 29.8+/-1.85 ng 20 hydroxyecdysone equiv. per mg tissue ovaries from LD females and 13.1+/-3.55 ng 20E equiv./mg in ovaries from SD females. The sum of RIA-positive materials in newly laid eggs was more than three times higher in non-diapause eggs than in diapause eggs. Ecdysteroids present in egg extracts comprised about 85% polar conjugates, 5% free ecdysteroids and 10% non-hydrolyzable metabolites. On the other hand, after diapause termination the amount of ecdysteroids increased drastically. Also, the composition of ecdysteroids differed from that observed during diapause and became comparable to that of non-diapause eggs. The significant differences in the ecdysteroids between non-diapause and diapause eggs may suggest the possible involvement of these compounds in the control of embryonic diapause of this locust. PMID- 12770070 TI - Effects of temperature on the establishment potential of the predatory mite Amblyseius californicus McGregor (Acari: Phytoseiidae) in the UK. AB - Amblyseius californicus was introduced into the UK in the early 1990s as a biocontrol agent against glasshouse red spider mite Tetranychus urticae. This study investigated the effects of temperature on the establishment potential of A. californicus in the UK in the light of recent reports of their successful overwintering outside of glasshouse environments. The developmental thresholds were 9.9 and 8.6 degrees C respectively using simple and weighted linear regression. Using the day-degree requirement per generation calculated by weighted regression (143 day-degrees) in combination with climate data, it was estimated that up to seven generations would be possible annually outdoors in the UK. Non-diapausing adult females froze at -22 degrees C, with 100% mortality after reaching their freezing temperature. Up to 90% of mites died before freezing after short exposures to low temperatures. Significant acclimation responses occurred; 90% of acclimated individuals survived 26 days exposure at 0 degrees C and 11 days at -5 degrees C (acclimated mites were reared at 19 degrees C, 6L:18D followed by 1 week at 10 degrees C, 12L:12D). Non-diapausing adult females survived over 3 months outdoors in winter under sheltered conditions and oviposition was observed. The experimental protocol used in this study is discussed as a pre-release screen for the establishment potential of other Amblyseius species, and similar non-native biocontrol agents. PMID- 12770071 TI - Adipokinetic hormone enhances laminarin and bacterial lipopolysaccharide-induced activation of the prophenoloxidase cascade in the African migratory locust, Locusta migratoria. AB - Lom-AKH-I enhances the activation in vivo of prophenoloxidase in the haemolymph of the African migratory locust, Locusta migratoria, in response to challenge with laminarin. AKH does not influence the speed or initial magnitude of the phenoloxidase response to laminarin, but prolongs the period of activation of the enzyme in a dose-dependent manner. Injections of preparations of bacterial lipopolysaccharide (LPS) do not activate prophenoloxidase in vivo, but co injection of Lom-AKH-I with commercial preparations of LPS from Klebsiella pneumoniae, Escherichia coli, or Shigella flexneri (but not one from Pseudomonas aeroginosa) results in dose-dependent increases in the levels of phenoloxidase that persist in the haemolymph for several hours. It is argued that the effects of AKH on phenoloxidase activation in locusts described here are, at least in part, related directly to changes in lipid metabolism brought about by the hormone. PMID- 12770072 TI - Cholesterol efflux from larval Manduca sexta fat body in vitro: high-density lipophorin as the acceptor. AB - The objective of this study was to characterize the transfer of cholesterol from Manduca sexta larvae fat body to high-density lipophorin. [(3)H]-Cholesterol labeled fat body was incubated with lipophorin under different conditions and cholesterol transfer was determined. Transfer rate exhibited a hyperbolic dependency on lipophorin concentration with an apparent K(m) of 3.6 mg/ml, which is consistent with either an aqueous diffusion mechanism of cholesterol transfer or a receptor-mediated process. Several results, including the high K(m), the high activation energy, and the lack of Ca(2+) dependence favor aqueous diffusion model. In addition, anti-lipid transfer particle antibodies had only a small inhibitory effect, suggesting it is not involved in cholesterol transfer. However, the transfer was inhibited in the presence of suramin, which would be consistent with a receptor-mediated process. The effects of suramin may be complex because it can change membrane properties when bound to the lipophorin receptor and affect the rate of cholesterol desorption. The preponderance of data suggests that the export of cholesterol from fat body to lipophorin follows a simple aqueous diffusion pathway. Although we cannot completely exclude some contribution from a receptor-mediated pathway, it seems that if such a pathway were present, it represents a minor route. PMID- 12770073 TI - Maternal reproductive decisions are independent of feeding in the black bean aphid, Aphis fabae. AB - Aphids are phloem feeders and an important assumption has been that reproduction is initiated only after phloem ingestion. Here we investigate the plant tissue location of parturition cues in winged and wingless, summer virginoparae and autumn migrants (gynoparae) of the black bean aphid, Aphis fabae. These seasonal forms have different host preferences. Using electrical penetration graph (EPG: to observe activity of the mouthparts) and video-monitoring procedures we demonstrate that the time to first parturition after host-plant contact is significantly shorter than the time to first registered phloem contact in the summer winged form. In gynoparae, the time to first parturition does not significantly differ from time to first phloem contact but is shorter than time to first phloem ingestion. Times to first parturition, first registered phloem contact and first phloem ingestion do not differ significantly in the summer wingless form. Simultaneous EPG and video recording procedures show that a high proportion of individuals of all morphs (45-70%) initiate reproduction before sustained phloem activities (salivation/ingestion). The only behaviours that all individuals demonstrate before first parturition are 'non-penetration' (aphid on plant surface with mouthparts outside plant) and stylet 'pathway activity', including secretion of gelling saliva and penetration of the non-vascular (epidermis and mesophyll) cells. A short period of penetration of the peripheral plant tissues (five cell punctures per individual) by aphids tethered and monitored by EPG decreases the time to first parturition of the winged summer form when subsequently placed on a Parafilm sachet containing 15% sucrose solution. This treatment also significantly increases the incidence of reproduction and individual reproductive output of gynoparae over a 24-h period. No detectable effects of tissue penetration on subsequent reproductive output are observed in the wingless summer form. Additionally, EPGs reveal that a number of aphids of all morphs display xylem ingestion, which occurs predominantly before initiation of phloem feeding but is not necessary to initiate parturition. It is concluded that aphids are likely to detect parturition cues during stylet punctures of cells within peripheral tissue layers, before reaching the phloem vessels. PMID- 12770074 TI - Electrophysiological and morphological characterization of the medulla bilateral neurons that connect bilateral optic lobes in the cricket, Gryllus bimaculatus. AB - The medulla bilateral neurons (MBNs) in the cricket brain directly connect two optic lobes and have been suggested to be involved in mutual coupling between the bilateral optic lobe circadian pacemakers. Single unit analysis with intracellular recording and staining with Lucifer Yellow was carried out to reveal morphology and physiology of the MBNs. Neurons having a receptive field in the rostral part of the compound eye showed greater response and a higher sensitivity to light than those having receptive fields in the ventro-caudal or dorsal portions. The MBN showed diurnal change in their responsiveness to light; the light-induced response in the night was about 1.3, 5 and 2 times of that in the day in MBN-1s, -3s and -4s, respectively. These results suggest that the MBNs mainly encode the temporal information by the magnitude of light-induced responses. The differences in magnitude of light-induced responses and of daily change in photo-responsiveness among MBNs may suggest that each group of MBNs plays different functional role in visual and/or circadian systems. PMID- 12770075 TI - The control of the proventriculus in the honeybee (Apis mellifera carnica L.) I. A dynamic process influenced by food quality and quantity? AB - The control of crop emptying in foraging honeybees was investigated in individuals trained to collect defined amounts of sugar solutions. Following feeding, they were dissected after fixed periods of time in order to measure crop content and haemolymph sugar titers. Between feeding and dissection, the metabolic rate of every investigated forager was measured using open-flow respirometry, so as to assess the effects of both food quality (concentration, molarity and viscosity of the fed sugar solution) and food quantity on the transport rate through the proventriculus. The sugar transport rate through the proventriculus was observed to be mainly dependent on the metabolic expenditure of the individual. Bee foragers were able to precisely adjust the sugar transport rate to their metabolic rates, but under certain conditions, an excess of sugars was transported through the proventriculus, more than needed to cover the bee's energetic demands. This excess depended on the nutritive value and quantity of the fed sugar solution, and on the time after feeding. It did not depend on the metabolic rate of the bee, the molarity, or the viscosity of the fed sugar solution. As long as the bees did not exhaust their crop contents, the haemolymph sugar titers were unaffected by this excess amount transported, by the time after feeding, the concentration and the viscosity of the fed sugar solution. For all feeding conditions assayed, the haemolymph trehalose titer remained constant, while the titers of other haemolymph sugars varied. It is suggested that the trehalose concentration in the haemolymph is regulated in honeybees, and that it represents the controlled variable in the feedback loop responsible for the transport rate through the proventriculus. PMID- 12770076 TI - A geometric analysis of nutrient regulation in the generalist caterpillar Spodoptera littoralis (Boisduval). AB - Nutritional regulatory responses of the generalist caterpillar Spodoptera littoralis (Boisduval) were explored, in choice and no-choice experiments, using the Geometric Framework. In the choice experiment, newly moulted final instar larvae were provided with one of three protein-biased foods (PB-food: p35:c7, p28:c5.6 or p21:c4.2) and one of three equal protein-carbohydrate ratio foods (ER food: p21:c21, p16.8:c16.8 or p12.6:c12.6). On five of the nine treatments, caterpillars independently regulated protein and carbohydrate intake to a mixture of 57 and 43%, respectively. However, when the concentration of the ER-food decreased and that of the paired PB-food increased, caterpillars progressively abandoned regulation and ate more of the PB-food. Despite these regulatory differences, performance (survivorship, growth and development) was similar across all nine treatments. In the no-choice experiment, caterpillars were given one of five foods (p35:c7, p28:c14, p21:c21, p14:c28 and p7:c35). Results indicated that caterpillars moved to a point in protein-carbohydrate space that was consistent with the Equal Distance Rule of compromise, a pattern previously predicted for generalist feeders. The insects on the two extreme foods, p35:c7 and p7:c35, showed reduced pupal mass and longer development, respectively. There was also strong evidence for post-ingestive regulation of nutrient utilisation, notably for protein. PMID- 12770077 TI - For uptake of yolk precursors, epithelial cell-oocyte gap junctional communication is required by insects representing six different orders. AB - For uptake of vitellogenin protein into nascent yolk spheres, communication through open gap junction channels between the follicle epithelium and oocyte is required by six different insects representing six different orders. It was recently shown in the hemipteran, Oncopeltus fasciatus, that endocytic uptake of yolk protein resulting in the formation of nascent yolk spheres depended upon an intact epithelium communicating with the oocyte through patent gap junctions. Following treatment with octanol, which down-regulated gap junctions below the level of dye coupling, vitellogenin uptake was terminated. Yet, for another hemipteran, Dysdercus intermedius, it has been shown that yolk spheres can form even when all epithelial cells have been stripped from the oocyte. To determine if the mechanism seen in Oncopeltus is present in other insects, we utilized the same techniques to study nascent yolk sphere production in a dipteran, Drosophila melanogaster, a lepidopteran, Actias luna, a hymenopteran, Xylocopa virginica, a coleopteran, Tenebrio molitor and an orthopteran, Acheta domesticus. In each of these, when gap junctions were down-regulated yolk uptake quickly stopped. That six different insects from six different orders all required a gap junctionally transmitted chemical signal of epithelial cell origin suggests that this mechanism is widespread throughout the insects. PMID- 12770078 TI - Metabolic and symbiotic interactions in amino acid pools of the pea aphid, Acyrthosiphon pisum, parasitized by the braconid Aphidius ervi. AB - Aphidius ervi Haliday (Hymenoptera, Braconidae) is an endophagous parasitoid of the pea aphid, Acyrthosiphon pisum (Harris) (Homoptera, Aphididae). This parasitoid strongly redirects host reproduction and metabolism to favour nutrition and development of its juvenile stages. Parasite-regulated biosynthesis and mobilization of nitrogen metabolites determine a significant increase of host nutritional suitability. The aim of the present study was mainly to investigate the temporal changes of A. pisum amino acid pools, as affected by A. ervi parasitism, and to assess the role of the aphid bacterial endosymbiont Buchnera in determining the observed changes. In parasitized aphids, we observed a very significant increase in total free amino acids, compared with synchronous non parasitized controls, starting from day 4 after parasitization (+51%). This trend culminated with more than doubling the control value (+152%) on day 6 after parasitization. However, a significant "parasitism" effect was observed only for 10 of the 28 amino acids detected. Tyrosine accumulation was the most prominent parasitoid-induced alteration, with a fourfold increase over control levels registered on day 6. In parasitized hosts, the amino acid biosynthetic capacity of Buchnera was unaltered, or even enhanced for the phenolic pool, and contributed greatly to the definition and maintainance of host free amino acid pools. The hypertyrosinemic syndrome was not dependent on food supply of the aromatic nucleus but was induced by parasitism, which likely enhanced the aromatic shuttle mediating phenylalanine transfer from bacteria to the host tissues, where tyrosine conversion occurs. This process is likely associated with a selective disruption of the host's functions requiring tyrosine, leading to the remarkable accumulation of this amino acid. The possible mechanisms determining these parasitism-induced host alterations, and their nutritional significance for the developing parasitoid larva, are discussed. PMID- 12770079 TI - Effects of the polydnavirus of Cotesia congregata on the immune system and development of non-habitual hosts of the parasitoid. AB - Polydnaviruses (PDV) are obligate mutualistic symbionts found in association with some groups of parasitic Hymenoptera. In these groups, they suppress the immune response of the parasitoid's host and are required for successful parasitoid reproduction. Several PDV effects have been described in different experimental systems, but no clear picture of PDV mode of immunosuppression has emerged. No study to date has directly tested if PDV modes of action are evolutionarily conserved or divergent among parasitoid taxa within the Ichneumonoidea. We hypothesize the divergence in PDV mode of immunosuppression can be detected by identifying points of divergence in the immune response of different host species to PDV from one parasitoid species. This study tests the effects of purified PDV from Cotesia congregata on the immune response of three larval lepidopteran species that naturally are hosts of parasitoid species that differ in taxonomic relatedness to C. congregata. Here we demonstrate that despite associations with distantly related parasitoids (Ichneumonidae and Braconidae), Manduca sexta and Heliothis virescens showed similar patterns of increased glucose dehydrogenase (GLD) activity, suppressed cellular encapsulation in vitro, and increased time to pupation. In contrast, Lymantria dispar showed no response to C. congregata PDV across any of the parameters measured, even though it has an evolutionary association with several parasitoids closely related to C. congregata and within the Microgastrinae. The PDV immunosuppression in H. virescens and M. sexta does not correlate with host molecular phylogeny either. The suborganismal effects shown in M. sexta and H. virescens translated into significantly reduced pupation success in M. sexta only. Results demonstrate that while some PDV modes of immunosuppression in hosts may be divergent, others may be conserved across broad host groups. PMID- 12770080 TI - Functional specialisation and polyphenism in aphid olfactory sensilla. AB - Electroantennogram (EAG) responses to the aphid sex pheromone components, (1R,4aS,7S,7aR)-nepetalactol and (4aS,7S,7aR)-nepetalactone, and a plant volatile, (E)-2-hexenal, were investigated at three different positions (5/6th, 4/5th and 3/4th inter-segmental regions) along the antennae of four different morphs in two host-alternating aphid species, Aphis fabae Scopoli and Rhopalosiphum padi (L.). Position-dependent and morph-specific EAG responses were elicited in both species. The nepetalactol and nepetalactone isomers elicited large EAG responses at all three recording positions in males of both species, such that primary rhinaria as well as secondary rhinaria appeared to respond. Asexual female morphs showed relatively smaller EAG responses to these compounds. The secondary rhinaria, which have been reported as sex pheromone receptors in males, were not very different in their number and distribution between gynoparae and alate virginoparae, but the gynoparae showed significantly larger EAG responses to nepetalactol and nepetalactone. The alate virginoparae showed EAG responses that were similar to those of apterous virginoparae, which lack secondary rhinaria. Taking the EAG response profiles together with the distribution of the secondary rhinaria, it is suggested that the function of secondary rhinaria differs between the morphs. Secondary rhinaria appear to detect sex pheromone components in males and gynoparae but not in the alate virginoparae. If they are functional in the latter morph, they are likely to play a role in detecting specific, but as yet unknown, volatile compounds. Some 30 plant volatiles were tested but none evoked an EAG response that could be allocated to the secondary rhinaria. In contrast to the very different EAG response profiles to the pheromone compounds between morphs, EAG responses to (E) 2-hexenal were similar in all forms and both species. These findings suggest that this plant volatile was detected only by the two primary rhinaria, which are common to all morphs. The present study showed that EAG responses were not a simple summation of receptor potentials between recording and reference electrodes in aphids. The localised distribution pattern of olfactory receptor neurones around the recording electrode was also likely to contribute to the EAG responses. PMID- 12770081 TI - Regulation of phospholipase A(2) activity in cockroach (Periplaneta americana) fat body by hypertrehalosemic hormone: evidence for the participation of protein kinase C. AB - Phospholipase A(2) (PLA(2)) associated with the membrane fraction of trophocytes from Periplaneta americana fat body increases by as much as 100% when the cells are incubated with hypertrehalosemic hormone (HTH-II). Activation with HTH-II is approximately halved by inclusion of the PKC inhibitor sphingosine in the incubation medium. Because activation of PLA(2) by HTH-II is blocked by the GDP analogue GDP-beta-S, and the unactivated enzyme is activated by the GTP analogue GTP-gamma-S it is likely that a G protein is involved in activation of the enzyme. Activation of PLA(2) was also achieved by treating the trophocytes with the synthetic diacylglycerol 1-oleoyl-2-acetylglycerol in the presence of thapsigargin. This supports the view that protein kinase C is also involved in the activation process. PMID- 12770082 TI - Lipid metabolism during sequential gonotrophic cycles in large and small female Aedes aegypti. AB - The lipid metabolism was investigated during six gonotrophic cycles of Aedes aegypti. Females of constant body size were analyzed for their total lipid content: large females with a body size of 41.06 (wing length cubed) and small females with 15.63. Their lipid contents at eclosion were compared to lipid values after two days of sugar-feeding, shortly before a blood meal, after oviposition, of their total egg batches, and again before the next blood meal, with intermittent access to sugar for two days for six gonotrophic cycles.Large females transferred most of their pre-blood meal lipid into the ovaries. Their low lipid content after oviposition was restored by synthesis from intermittent sugar meals. After the third gonotrophic cycle, they withheld more and more of the resynthesized lipid in their fat body, thus gradually reducing their fecundity. Since blood consumption was not altered significantly during these six cycles, age-related reduction of fecundity was clearly caused by limitations of yolk lipid.Small females transferred a considerably smaller, but constant segment of sugar-derived lipids to the ovaries. In both size classes, lipid content per oocyte was constant throughout all cycles with 9 mcal/oocyte in large and 7 mcal/oocyte in small females. Total fecundity reached 450 eggs in large and 280 eggs in small females. Large females that were maintained on water without sucrose took large blood meals from which part of the yolk lipid was synthesized. Extrapolations suggest that only one or two additional gonotrophic cycles would be possible without additional carbohydrate sources, despite lipogenesis from blood protein. PMID- 12770083 TI - Absorption and storage of phosphorus by larval Manduca sexta. AB - The role of phosphorus (P) in numerous important biological structures, coupled with the observation that P-content of many insect foods is disproportionately low, suggests that P may be a critical nutrient for growing insects - however, the few studies examining the effects of dietary P on insect performance have generally found only weak relationships. This mismatch may be reconciled by understanding the physiological mechanisms by which insects handle P. Here we describe P processing by larvae of Manduca sexta. When given un-manipulated leaves of a common host plant, Datura wrightii, fifth-instar larvae retained about 85% of P consumed; when given P-enriched leaves larvae retained only 25% of P consumed. Analysis of gut concentrations of P at four sites along the digestive tract, and in leaves and feces, indicates that the rectum is the primary site of P transport between the gut and body and that differences in P retention may be accounted for by differential rates of rectal P transport. Larvae given P enriched leaves also showed an eightfold increase in the concentration of P in the hemolymph, primarily as alpha-glycerophosphate - but only a 12% increase in the concentration of P in body tissues, suggesting that hemolymph plays a central role in storage and buffering of P. PMID- 12770084 TI - The effects of amiloride on the labellar taste receptor cells of the fleshfly Boettcherisca peregrina. AB - Amiloride is known to inhibit the taste response of vertebrates to salt by blocking the amiloride-sensitive sodium channel. In this study, we investigated electrophysiologically the effect of amiloride on the taste response of the fleshfly Boettcherisca peregrina. When 0.5 mM amiloride was included in taste solutions, the response of the salt receptor cell (salt response) to sodium chloride (NaCl) was not depressed but those of the sugar receptor cell (sugar responses) to sucrose, glucose, fructose, L-valine (L-Val) and L-phenylalanine (L Phe) were strongly depressed. An inhibitory effect of amiloride on the concentration-response relationship for both sucrose and L-Phe was clearly revealed, but not at high concentrations of sucrose. After pretreatment of a chemosensory seta with 0.15 mM amiloride for 10 min, the salt response to NaCl was not affected. On the other hand, the sugar responses to sucrose, fructose, L Val and L-Phe were depressed just after amiloride pretreatment. The sugar response to adenosine 5'-diphosphate (ADP) mixed with 0.5 mM amiloride was not depressed, but the response to ADP alone was depressed after amiloride pretreatment. It was therefore observed that amiloride depressed the responses to all stimulants that react with each of the receptor sites of the sugar receptor cell. PMID- 12770085 TI - Temperature acclimation in overwintering nymphs of a cockroach, Periplaneta japonica: walking on ice. AB - Nymphs of the Yamato cockroach, Periplaneta japonica, showed a seasonal change in the ability to move at low temperature. Laboratory-reared nymphs buried in ice for 15 min-3 h took about 600 s to get up at 25 degrees C after being placed with their dorsal surface on the floor of a Petri dish. The mean time to get up at 25 degrees C (recovering time) after 15 or 60-min burial in ice was relatively long in early autumn, decreased rapidly in autumn, reached a minimal level in winter (<100 s) and increased in spring. Temperature was responsible for this seasonal change in physiology, and neither photoperiod nor diapause status was important. Recovering time depended on the acclimation temperature and its duration. For a 10-day exposure in a range of 0-25 degrees C, the lower the temperature the shorter the recovering. Longer periods of acclimation at these temperatures tended to shorten recovering time. The acquisition of the ability to move on ice is a seasonal phenomenon unique to P. japonica. No comparable response was found in three other cockroaches including two sub-tropical species (Periplaneta americana and Opisthoplatia orientalis) and a temperate species (Blattela nipponica). PMID- 12770086 TI - A novel regulatory mechanism for amino acid absorption in lepidopteran larval midgut. AB - A number of methyl and ethyl esters of naturally occurring amino acids exert a potent stimulatory effect on the cotransport system responsible for the absorption of most essential amino acids along the midgut of the silkworm Bombyx mori. L-Leucine methyl ester (Leu-OMe), one of the most effective activators, induces a large increase of the initial rate of leucine uptake in midgut brush border membrane vesicles (BBMV) from the anterior-middle (AM) region, and a small effect in BBMV from the posterior (P) region. Nonetheless, the methyl ester causes in both regions a relevant K(+)-, Deltapsi- and pH-independent increase of the intravesicular accumulation of the amino acid. The activation by Leu-OMe proves that amino acid absorption can be modulated all along the B. mori larval midgut and that the AM region, where the ability to transport and concentrate the substrate is very low, is more susceptible than the P region. Leucine uptake in AM-BMMV can be activated by amino acid methyl esters with definite structural requisites, with the following order of potency: L-leucine>L-phenylglycine>L methionine>L-phenylalanine>L-norleucinez.Gt;L-isoleucine. The activation is stereospecific and occurs also with some ethyl esters (e.g. leucine and phenylalanine). No activation was observed with esters of amino acids with short hydrophobic or polar side-chains. The activation mechanism here described plays a fundamental role in larval growth since silkworms reared on artificial diets supplemented with leucine or methionine methyl esters reach maximum body weight 12-18 h before control larvae and spin cocoons with a larger shell weight. This novel regulatory mechanism of an amino acid transport protein appears to be widespread among lepidopteran larvae. PMID- 12770088 TI - Lipid melting and cuticular permeability: new insights into an old problem. AB - The idea that the physical properties of cuticular lipids affect cuticular permeability goes back over 65 years. This proposal has achieved textbook status, despite controversy and the general lack of direct supporting evidence. Recent work supports the standard model, in which lipid melting results in increased cuticular permeability. Surprisingly, although all species studied to date can synthesize lipids that remain in a solid state at environmental temperatures, partial melting often occurs due to the deposition of lipids with low melting points. This will tend to increase water loss; the benefits may include better dispersal of lipids or other compounds across the cuticle or improved communication via cuticular pheromones. In addition, insects with high melting point lipids are not necessarily less permeable at low temperatures. One likely reason is variation in lipid properties within the cuticle. Surface lipids differ from one region to another, and biophysical studies of model mixtures suggest the occurrence of phase separation between melted and solid lipid fractions. Lipid phase separation may have important implications for insect water balance and chemical communication. PMID- 12770089 TI - Extended in vitro culture of Microplitis croceipes teratocytes and secretion of TSP14 protein. AB - Teratocytes, cells which originate from the serosal membrane of some Braconidae and Scelionidae, can be found in the hemocoel of permissive hosts during part or all of the developmental time of the parasitoid larva. Teratocytes from Microplitis croceipes are known to secrete biologically active proteins, which contribute to developmental arrest and failure to pupate of Heliothis virescens larvae. One such protein, which has a molecular weight of approximately 14 kDa is called TSP14. The presence of parasitoid larvae is essential to maintain teratocytes under in vitro conditions with protein-free EX-CELL 400. The teratocyte viability was maintained in vitro for at least 12 days in the presence of larvae when medium was exchanged every three days. Western blots show that TSP14 was secreted during the entire period of exchanges. In the absence of parasitoid larvae, teratocyte viability was only 30% by day 6 and no TSP14 had been secreted. In the absence of parasitoid larvae, teratocytes maintained in vitro in EX-CELL 400 medium supplemented with 10% FBS remained viable for at least nine days and secreted TSP14 for at least six days. This suggests that parasitoid larval secretions are sufficient but not uniquely essential to maintain teratocyte viability. Parasitoid larvae maintained in the absence of teratocytes did not secrete TSP14 and their secretory products did not inhibit pupation of H. virescens larvae. PMID- 12770090 TI - Possible involvement of ecdysteroids in photoperiodically induced suppresion of ovarian development in a Japanese strain of the migratory locust, Locusta migratoria. AB - In a Japanese population of Locusta migratoria, adult females become reproductively inactive under crowding and long days (LD) and reproductively active under crowding and short days (SD). The identity and titre of ecdysteroids in the haemolymph and ovaries from adult females reared under SD and LD were investigated by RIA/HPLC. The effects of exogenous juvenile hormone (JH) III treatments on the termination of such reproductive arrest and ecdysteroid contents in LD females were also examined. In general, ecdysteroid titres in both haemolymph and ovaries were significantly higher in reproductively active SD females than in reproductively inactive LD females. A clear difference was also observed in oocyte growth between SD and LD individuals. JH III applications (three consecutive topical applications, 150 &mgr;g per insect per day from day 3) stimulated ovarian development in LD females and significantly increased the haemolymph and ovarian ecdysteroids to a level comparable to that of reproductively active SD adult females. PMID- 12770091 TI - Differential sensitivity of two insect GABA-gated chloride channels to dieldrin, fipronil and picrotoxinin. AB - In the central nervous system of both vertebrates and invertebrates inhibitory neurotransmission is mainly achieved through activation of gamma-aminobutyric acid (GABA) receptors. Extensive studies have established the structural and pharmacological properties of vertebrate GABA receptors. Although the vast majority of insect GABA-sensitive responses share some properties with vertebrate GABAA receptors, peculiar pharmacological properties of these receptors led us to think that several GABA-gated chloride channels are present in insects. We describe here the pharmacological properties of two GABA receptor subtypes coupled to a chloride channel on dorsal unpaired median (DUM) neurones of the adult male cockroach. Long applications of GABA induce a large biphasic hyperpolarization, consisting of an initial transient hyperpolarization followed by a slow phase of hyperpolarization that is not quickly desensitized. With GABA, the transient hyperpolarization is sensitive to picrotoxinin, fipronil and dieldrin whereas the slow response is insensitive to these insecticides.When GABA is replaced by muscimol and cis-4-aminocrotonic acid (CACA) a biphasic hyperpolarization consisting of an initial transient hyperpolarization followed by a sustained phase is evoked which is blocked by picrotoxinin and fipronil. Exposure to dieldrin decreases only the early phase of the muscimol and CACA induced biphasic response, suggesting that two GABA-gated chloride channel receptor subtypes are present in DUM neurones. This study describes, for the first time, a dieldrin resistant component different to the dieldrin- and picrotoxinin-resistant receptor found in several insect species. PMID- 12770092 TI - Antennal resolution of pulsed pheromone plumes in three moth species. AB - Male antennae of Cadra cautella, Pectinophora gossypiella, and Spodoptera exigua were presented with 20-ms-duration pulses of their two-component pheromone at rates of 1 to 33 Hz. Fourier analyses of electroantennograms resolved the temporal structure of trains of pheromone filaments delivered at up to 33 Hz for C. cautella and S. exigua and 25 Hz for P. gossypiella. Pheromone components tested separately for each species were generally equivalent in filament resolution to complete blends. Ambient temperatures of 18, 23 and 28 degrees C affected filament resolution only slightly, with poorer ability to discriminate rapidly pulsed signals at 18 degrees C. The question of how, or indeed if, such frequencies are conserved beyond the peripheral nervous system, remains. PMID- 12770093 TI - Responses in highly selective sensory neurons to blends of pheromone components in the moth Agrotis segetum. AB - Pheromone detecting sensory neurons in moths are known to be highly sensitive and selective. Female-emitted sexual pheromones are normally mixtures of a few to several components. However, not much is known about how receptor neurons respond to blends of compounds. In the present study we investigated how four physiological types of pheromone component-selective neurons responded to binary mixtures or to the complete blend in the turnip moth Agrotis segetum. We found that responses to mixtures only rarely differed from that to the excitatory component alone. The mixture interactions were exclusively suppressive and occurred only at high concentrations. Therefore we conclude that the, in A. segetum, commonly observed mixture interactions observed in higher brain centra are mainly the result of central nervous processing and that information about the pheromone components reaches the antennal lobes virtually unaltered. In addition, we found a physiological type of receptor neuron, responding selectively to one of the female-emitted pheromone components, that has previously not been observed in the Swedish population. PMID- 12770094 TI - Secretions of stingless bees: cephalic secretions of two Frieseomelitta species. AB - Gas chromatographic and mass spectrometric analysis of the volatile compounds in the mandibular glands of Frieseomelitta varia and the heads of Frieseomelitta silvestrii have revealed relatively simple or more complex mixtures of volatile oxygenated compounds. 2-Alkanols were found to be important components for both species. In F. varia the composition was essentially the same in samples from two widely separated areas and there were small differences between callows and mature, foraging bees. The first electroantennographic studies on stingless bees, using hexane extracts of heads of both species, have demonstrated a response in the workers' antennae to these, as well as to the pure compounds 2-heptanol and 2 nonanol. PMID- 12770095 TI - Oviposition deterrence associated with larval frass of the Japanese pine sawyer, Monochamus alternatus (Coleoptera: Cerambycidae). AB - To determine the effect of larval frass of Monochamus alternatus on oviposition preference of the female adults, three tests were performed in the laboratory. Individual females were provided with a frass-coated, Pinus densiflora bolt and an untreated bolt simultaneously and were allowed to oviposit for 24 h. They deposited a significantly smaller number of eggs on frass-coated bolts. The females supplied with frass-coated bolts deposited no eggs on them, indicating that the larval frass deterred the females from oviposition. When individual females were provided simultaneously with a pine bolt applied with a methanol extract of larval frass and another bolt applied with methanol alone, they deposited a significantly smaller number of eggs on frass extract-applied bolts for 24 h. These results suggest the presence of a putative oviposition deterrent in larval frass of M. alternatus. Each test was replicated 19-20 times using different females. PMID- 12770096 TI - Taste detection of phytoecdysteroids in larvae of Bombyx mori, Spodoptera littoralis and Ostrinia nubilalis. AB - A number of plants produce significant amounts of phytoecdysteroids that can disrupt the hormonal levels of insects feeding upon them. Insects equipped with taste receptors sensitive to phytoecdysteroids are able to avoid such plants. How common is this strategy? By recording from the lateral and medial sensilla styloconica in two polyphagous species (Ostrinia nubilalis and Spodoptera littoralis) and in a monophagous species (Bombyx mori), we tested whether the receptors could detect three commonly occurring phytoecdysteroids 20 hydroxyecdysone (20E), ponasterone A (ponA) and ecdysone (E). In B. mori, 20E and ponA elicited dose-dependent responses with a threshold of 1 &mgr;M only in the medial sensilla. In O. nubilalis, 20E, E and ponA elicited responses at threshold of 1 &mgr;M in both sensilla. In S. littoralis, 20E elicited responses with a threshold of 10 &mgr;M in both sensilla. By means of behavioural choice tests, we show that 20E is an effective feeding deterrent for O. nubilalis and S. littoralis first instar larvae. This suggests that the perception of phytoecdysteroids is more common among phytophagous lepidoptera than previously thought, although their toxicity or antifeedancy varies between species. PMID- 12770097 TI - Effect of age, diet, diapause and juvenile hormone on oogenesis and the amount of vitellogenin and vitellin in the twospotted stink bug, Perillus bioculatus (Heteroptera: pentatomidae). AB - Vitellogenic oocytes from Perillus bioculatus have two native vitellins, Vt1 and Vt2, with molecular masses of 553 and 228 kDa, respectively. The hemolymph contains a major vitellogenin, Vg, with a molecular mass of 528 kDa that consists of three apoproteins with masses of 177, 84 and 59 kDa, respectively. Antibodies to purified Vt2 reacted with ovary extracts, egg extracts and female hemolymph, but not with male hemolymph in immunodiffusion tests. Western blots showed that anti Vt2 reacted with both Vt1, Vt2 and with Vg. Vitellogenesis starts at an ovarian score of 12 at 2.4 days after emergence. The first cycle of egg development is completed in ovaries with a score of 112 at 7.7 days. During this 5.3 day period, the ovaries of a single female incorporated 1833 &mgr;g of protein to form vitellin. Vitellogenin levels start to increase in females 2.5 days after emergence and reached 17.8 &mgr;g/&mgr;l by 5.5 days. After 5.5 days vitellogenin levels fluctuated between 9.7 and 19.9 &mgr;g/&mgr;l. Most diapausing females contained no ovarian follicles in the vitellarium and their hemolymph contained less than 1 &mgr;g/&mgr;l of vitellogenin. Treating diapausing females with 1 &mgr;g of JH III increased vitellogenin levels over 120 fold. Insects maintained on a liver-based artificial diet had lower vitellogenin levels than the controls at all sample times and did not show an increase in vitellogenin concentration until 11.5 days. Treating insects on the artificial diet with 10 &mgr;g of JH III elevated vitellogenin levels to about a fourth of that found in prey-fed insects of a comparable age. This suggests that females fed the artificial diet have low levels of essential materials needed for vitellogenin production. PMID- 12770098 TI - Developmental rates of two congeneric parasitoids, Encarsia formosa and E. pergandiella (Hymenoptera: Aphelinidae), utilizing different egg provisioning strategies. AB - Endoparasitic Hymenoptera vary in the extent to which they provision their eggs and thus in the degree to which they appear to rely on their hosts for resources during embryonic development. In this study, developmental rates were examined in two congeneric parasitoid species, Encarsia formosa and E. pergandiella, that provision their eggs to different degrees. E. formosa eggs are much larger than E. pergandiella eggs. E. formosa eggs hatch significantly earlier than the eggs of E. pergandiella when deposited in 1st or 4th instar nymphs of a common whitefly host, Bemisia tabaci. Both species hatch earlier in 4th instar nymphs, but the delay in hatching in hosts parasitized as 1st instars is much greater in E. pergandiella. While E. formosa develops more rapidly to the 1st larval instar, E. pergandiella emerge as adults significantly earlier, though smaller, than E. formosa adults regardless of the host instar parasitized. These findings show that the extent of provisioning in the eggs of these wasps does not strictly determine their order of progression through different stages of development. PMID- 12770099 TI - Maintenance of soldier-producing aphids on an artificial diet. AB - In our efforts to understand the biology of soldier-producing aphids, we attempted to maintain them in the laboratory using a chemically defined artificial diet. The ability of 16 species from the subfamilies Eriosomatinae and Hormaphidinae, most of which are soldier-producing species, to survive on the artificial diet was examined. Some species neither fed nor grew on the diet, whereas other species accepted the diet, grew to some extent, and managed to produce a small number of short-lived offspring. Although they performed poorly on the diet in general, aphid performance was correlated with the stage in the life cycle and the developmental stadium in that aphids of the gall generation tended to accept the diet and survive on it, whereas aphids of the non-gall generation did not. Also, old insects tended to perform better on the diet than young nymphs. Notably, only one species, Tuberaphis styraci, a gall-forming aphid that produces 2nd instar sterile soldier, showed good performance on the diet. Insects collected from galls (generation G0) survived on the diet, grew well, and produced many progeny. Three successive generations (G1, G2 and G3) were produced on the diet. Developmental period, adult body size, and age of first reproduction were almost constant through G0, G1 and G2 whereas fecundity, adult longevity and daily offspring production declined as the generations proceeded. These results are comparable to previous studies in which pest aphids have been maintained on similar artificial diets for several generations. Therefore, it is suggested that the artificial-diet rearing system will provide a useful tool to investigate various biological aspects of the soldier-producing eusocial aphid, T. styraci. PMID- 12770100 TI - Hemolymph proteins in ticks. AB - In comparison to insects and Crustacea, our knowledge of the predominant hemolymph proteins in ticks is minimal. The hemolymph protein most studied in ticks has been vitellogenin (Vg). Vg is synthesized by the tick fat body after female adults obtain a blood meal, is released into the hemolymph and is absorbed by developing oocytes as vitellin (Vn). Much of what we know about Vg is from studies of Vn. In general, the carbohydrate, lipid and amino acid composition is similar to insects except that in the tick, Vg contains heme, most likely from the digestion of host hemoglobin. In the American dog tick, Dermacentor variabilis, Vg is comprised of two native proteins and seven subunits on SDS PAGE. Vg has been characterized in five tick species but the amino acid sequence is not yet available. Another predominant hemolymph protein, apparently a carrier protein (CP), has recently been studied in two tick species. This protein is found in the hemolymph of both male and females adults, in adult tissues outside of the hemolymph in some tick species, in coxal fluid of soft ticks and in whole body homogenates from eggs, larvae and nymphs. CP from the hard tick, D. variabilis, contains cholesterol, phospholipids, monoacylglycerides, triacylglycerides, free fatty acids, carbohydrate and heme. Under identical assay conditions, the analogous protein in the soft tick, Ornithodoros parkeri, did not contain heme. CP in the American dog tick consists of two subunits, one of which has 61% identity to the biliprotein, artemocyanin, from the fairy shrimp. CP is identical to a heme-lipoprotein (HeLp) from Boophilus microplus. The exact roles of CP and HeLp have not yet been fully determined, but they apparently are important in heme sequestration and as a storage depot for protein and lipid. Macroglobulin, lectin, antimicrobial, JH binding, JH esterase, and other tick hemolymph proteins are also discussed. PMID- 12770101 TI - The role of photoperiod and temperature in determination of summer and winter diapause in the cabbage beetle, Colaphellus bowringi (Coleoptera: Chrysomelidae). AB - The cabbage beetle, Colaphellus bowringi, is a short-day species undergoing an imaginal summer and winter diapause. Its photoperiodic response highly depends on temperature. All adults entered diapause at 24 degrees C) showed that the beetle could lose its sensitivity to photoperiod very rapidly. PMID- 12770102 TI - Nestmate recognition signals of the leaf-cutting ant Atta laevigata. AB - Behavioral tests with field colonies of Atta laevigata were performed in order to identify the source of the odors used in nestmate recognition. We tested the postpharyngeal (PPG) and mandibular glands (MG) as putative organs producing chemical signals for nestmate recognition. Chemical analyses of PPG were also undertaken. With a series of bioassays, we confirmed that nestmate recognition is based on cephalic odors and that these odors come mainly from the mandibular gland secretion. We show chemical evidence that odors from MG are dispersed all over the cuticle. Although odors from PPG elicited colony-specific behavioral responses, the types of behaviors they elicited differed from those of nestmate recognition of whole ants or MG extracts. PPG secretion was characterized by long chain alkanes and methyl branched alkanes of low volatility, whereas MG contained volatile ketones and alcohols. PMID- 12770103 TI - Effects of the Diadromus pulchellus ascovirus, DpAV-4, on the hemocytic encapsulation response and capsule melanization of the leek-moth pupa, Acrolepiopsis assectella. AB - DpAV-4 is a symbiotic ascovirus found in natural populations of the solitary endoparasitoid wasp Diadromus pulchellus. The female wasp injects this virus into the pupae of the leek-moth Acrolepiopsis assectella during oviposition. The ascovirus replicates in the pupal tissues and the consequent lysis of the cells occurs synchronously with egg hatching and the development of the wasp larva. We show here that encapsulation and capsule melanization were activated when minute nylon monofilaments were implanted into the hemocoel of non parasitized leek-moth pupae and that encapsulation and melanization were inhibited in pupae parasitized by D. pulchellus. When the pupae were infected by DpAV-4, melanization of the nylon monofilaments was abolished, but a capsule was still always formed. These results indicate that DpAV-4 is a free virus able to alter the defence system of the parasitized host so as to improve the development of the parasitoid wasp, D. pulchellus. PMID- 12770104 TI - Diapause development and acclimation regulating enzymes associated with glycerol synthesis in the Shonai ecotype of the rice stem borer larva, Chilo suppressalis walker. AB - Overwintering larvae of the Shonai ecotype of the rice stem borer, Chilo suppressalis, enter diapause in early September and terminate diapause at the end of October. Cold acclimation at 0 degrees C did not influence glycerol, trehalose or glycogen content in larvae collected on 22 September. Acclimation at 0 degrees C increased the glycerol content and reduced the glycogen content significantly in larvae collected on 2 October and 22 November compared with acclimation at 15 degrees C. These results indicate that overwintering larvae at different phases of diapause development respond differently to the low temperature stimulus for glycerol synthesis. Thus, we evaluated the metabolic rearrangements associated with glycerol synthesis during diapause development and after temperature acclimation. Larvae collected on 2 October were acclimated at 15 degrees C for 15 and 60 days. Some of those acclimated at 15 degrees C were then moved to 0 degrees C for 15 days. The larvae acclimated at 15 degrees C for 15 days were in deep diapause and accumulated little glycerol, while larvae acclimated at 15 degrees C for 60 days were nearly ready to emerge from diapause and accumulated glycerol at 155.5 &mgr;mol/g. When larvae acclimated to 15 degrees C for 15 days were transferred to 0 degrees C, glycerol accumulation was stimulated to the same extent (ca 140 &mgr;mol/g) as it was in larvae that were acclimated to 15 degrees C for 60 days and then transferred to 0 degrees C. These results indicate that low temperature has a cumulative effect on glycerol production in larvae at different phases of diapause development. Glycerol accumulation was accomplished by activation of glycogen phosphorylase and inhibition of fructose-1,6 bisphosphatase, and activation of enzymes associated with glycerol synthesis, mainly glyceraldehyde-3-phosphatase and polyol dehydrogenase with glyceraldehyde activity. PMID- 12770105 TI - Sequestration of host plant carotenoids in the larval tissues of Helicoverpa zea. AB - To determine the cause of the unique yellow coloration in mandibular glands of soybean-fed Helicoverpa zea larvae, the accumulation of carotenoids in various tissues of last instar larvae fed soybean, cotton and tomato foliage was quantified. Five carotenoids were detected in the foliage of all host plants but at significantly different concentrations. Xanthophylls rather than carotenes were most likely to accumulate in larval tissues. Carotenoids accumulated at different rates and some were significantly affected by larval diet. Highest levels of carotenoid accumulation, notably lutein, were detected in the testes, followed by midgut epithelium, fat body and integument. The midgut epithelium contained the greatest and the testes the least diversity of carotenoid types. Low levels of lutein were detected in both labial and mandibular glands. Tomato foliage had the highest carotenoid content and caterpillar tissues fed these leaves often had the highest amounts of carotenoid. However, the accumulation of carotenoids did not protect larvae from antibiotic effects of tomato foliage because these caterpillars had the highest mortality and slowest growth rates of all the three host plants. Transport and absorption of lipid and oxidative stress may be some reasons for differential carotenoid accumulation. PMID- 12770106 TI - Age-specific patterns in honeydew production and honeydew composition in the aphid Metopeurum fuscoviride: implications for ant-attendance. AB - The intensity of the mutualistic relationship between aphids and ants depends mainly on the composition and amount of honeydew. We used the model system Tanacetum vulgare-Metopeurum fuscoviride to study age-related differences in honeydew production and composition and its effect on the mutualism between M. fuscoviride and the ant Lasius niger. First and second instar larvae of M. fuscoviride produced only half of the amount of honeydew as older larvae or adults. There were, however, no differences between age classes in the total honeydew sugar concentration, which averaged approx. 80 &mgr;g sugar/&mgr;l honeydew. Honeydew sugar composition also did not differ between age classes, and melezitose was the dominant sugar (59% in all classes). The amino acid concentration, by contrast, increased significantly with aphid age, reaching 22.6 nmol per &mgr;l honeydew in adult M. fuscoviride. This increase was mainly caused by asparagine and glutamine, while there were no differences in the concentrations of the five other regularly detected amino acids and cystine, respectively. The intensity of ant-attendance was significantly lower in colonies of first and second instar larvae than in colonies of older age classes. Ant attendance correlated with the amount of honeydew produced, and not with the total amino acid concentration. PMID- 12770107 TI - Ultrastructural changes to the midgut of the black field cricket (Teleogryllus commodus) following ingestion of potato protease inhibitor II. AB - Ultrastructural changes to the midgut epithelium of nymphs of the black field cricket (Teleogryllus commodus) after ingestion of potato protease inhibitor II (PPI-II) (0.6% (w/v) in artificial diet) were determined by light and electron microscopy. Crickets fed diet containing PPI-II grew more slowly than those fed control diet and changes observed to the PPI-II-fed nymphs included reduction of midgut wall depth, vacuolisation of the epithelial cells, swelling of the microvilli, cellular protrusions into the midgut and eventual rupture of individual or small groups of epithelial cells. These changes were first seen 2 days after PPI-II ingestion. Complete disintegration of the midgut to the basement membrane was not seen during the 27-day observation period and repair and regeneration of pockets of epithelial cells was observed. Immunocytochemistry revealed that PPI-II was localised within the ectoperitrophic matrix space of the gut. The location of the peritrophic matrix was determined by labelling with wheat germ agglutinin (WGA), but no rupture of this structure was observed in PPI II-fed nymphs. PMID- 12770108 TI - Effects of parasitism by Asobara tabida (Hymenoptera: Braconidae) on the development, survival and activity of Drosophila melanogaster larvae. AB - The impact of parasitism by Asobara tabida on Drosophila melanogaster larval development, survival features and larval activity has been investigated using two strains of the parasitoid. The successful parasitism rate of the A1 strain was four times greater than that of the WOPV strain. Both strains induced equivalent mortality rates but hosts parasitized by A1 predominantly died as pupae. The time necessary for the host pupariation and emergence, and the larval weight at 72, 96 and 120 h post-parasitization were measured. Parasitized larvae exhibited longer periods of development and lower weights than controls, especially when parasitized by A1. These results suggest that hosts underwent physiological costs varying with respect to the outcome of the parasitic relationship. Of the parasitoid factors possibly responsible for these costs, we examined venoms for their impact on host mortality. Artificial injections of WOPV venoms induced higher mortality rates than did A1 venoms. Venoms were also found responsible for the induction of a transient paralysis, naturally occuring after parasitization. Again, the strongest effect was observed after parasitization by WOPV or injections of its venoms. This study gives new insights into the intriguing features of A. tabida and constitutes the first report of the paralysing properties of the venoms. PMID- 12770109 TI - Diapause in the egg parasitoid Trichogramma cordubensis: role of temperature. AB - The role of temperature in the induction of diapause in Trichogramma cordubensis (Hymenoptera: Trichogrammatidae), under controlled laboratory conditions, was investigated using Ephestia kuehniella Zeller (Lepidoptera: Pyralidae) eggs as hosts. Results indicate that prestorage temperatures and the duration of exposure of the parasitoids to these temperatures affected the induction of diapause. It was possible to induce diapause in prepupae of T. cordubensis by exposing the preimaginal stages (prior to the prepupal stage) to 10 degrees C for at least 30 days, but adults emerged without diapause when the duration of exposure was of only 10 or 20 days. Parasitoids failed to enter diapause when prestorage temperatures were 7 or 12 degrees C, regardless of the duration of exposure. However, at these two temperatures, preimaginal development of T. cordubensis was delayed, allowing short-term storage (40 days at induction temperatures followed by 30 days at 3 degrees C) by keeping parasitoids in quiescence without reducing the percentages of adult emergence. Good percentages of adult emergence after long-term low-temperature storage (30 or 40 days at 10 degrees C followed by six months at 3 degrees C) occurred only when T. cordubensis was in diapause. The long-term storage of parasitoids in diapause allows an enlargement in the mass rearing potentialities of this species for future biological control releases by allowing producers to stockpile the parasitoids for release in the field season. PMID- 12770110 TI - Identification and action of juvenile hormone III from sexually mature alate females of the red imported fire ant, Solenopsis invicta. AB - Analysis of extracts of hemolymph obtained from sexually mature alate females of Solenopsis invicta from monogyne colonies resulted in identification of juvenile hormone III (JH III). The average amount of JH III was 0.32+/-0.04 pmol/&mgr;molof hemolymph. Topical application of 0.038 pmol of JH III was sufficient to stimulate alates to shed their wings in the presence of the queen. The time in which alates were induced to dealate decreased linearly with increasing concentrations of JH III from 0.038 to 3.8 pmol. However, higher JH III concentrations deviated from linearity and did not reach dealation times comparable with those that occur after mating flights. Thus, it appears that the mechanism of dealation that occurs when female alates are out of the influence of their queen is different from the one associated with mating flights. Application of 0.42 &mgr;mol of precocene II inhibited dealation of alates in queenless colonies. However, this inhibition was reversed after applying 38 pmol JH III to precocene-treated alates. The sizes of corpora allata (CA) from sexuals treated with JH III did not differ from those of controls. However, the sizes of CA were reduced in alates treated with precocene II. The results indicated that JH was important to dealation. PMID- 12770111 TI - Triterpenoid saponins stimulate the sugar taste receptor cell through a G protein mediated mechanism in the blowfly, Phormia regina. AB - The blowfly has taste chemosensilla on the labellum. The sensory receptor cells in the chemosensillum are highly specialized for the tastes of sugar, salt and water, respectively. Previously we introduced chromosaponin I (CSI) and glycyrrhizin (GL), as sweet substances for the blowfly, Phormia regina. Application of these triterpenoid saponins induced feeding responses as well as impulses of the sugar taste receptor cell in the LL-type sensillum at a much lower concentration than that of sucrose. In the present paper, we show the involvement of G protein-mediated cascade in the CSI- and GL-responses as well as in sugar responses. CSI activates the sugar signal transduction cascade after penetrating through the membrane. On the other hand, GL exerts dual effects to stimulate the sugar signal transduction possibly by activating it inside the cell and also by interacting with the pyranose sugar receptor site. A non hydrolyzable G protein inhibitor guanosine 5'-O-(2-thiodiphosphate), GDPbetaS, markedly decreased the responses of the sugar receptor cell to the two triterpenoid saponins as well as the response to sucrose and fructose. These results suggest that CSI and GL are direct activators of G protein. PMID- 12770112 TI - Membrane conductances of principal cells in Malpighian tubules of Aedes aegypti. AB - Two-electrode voltage clamp (TEVC) methods were used to explore conductive transport pathways in principal cells, the dominant cell type in Malpighian tubules of the yellow fever mosquito. The basolateral membrane of principal cells had a voltage (V(bl)) of -85.1 mV in 49 principal cells under control conditions. Measures of the input resistance R(pc) together with membrane fractional resistance yielded estimates of the conductance of the basolateral membrane (g(bl) = 1.48 &mgr;S) and the apical membrane (g(a) = 3.13 &mgr;S). K(+) channels blocked by barium accounted for 0.94 &mgr;S of g(bl). Estimates of transference numbers yielded the basolateral membrane Na(+) conductance of 0.24 &mgr;S, leaving 0.30 &mgr;S (20%) of g(bl) unaccounted. The secretagogue db-cAMP (0.1 mM), a known activator of the basolateral membrane Na(+) conductance, significantly depolarized V(bl) to -65.0 mV and significantly increased g(bl) from 1.48 &mgr;S to 2.47 &mgr;S. The increase was blocked with amiloride (1 mM), a known blocker of epithelial Na(+) transport. The inhibition of metabolism with di-nitrophenol significantly depolarized V(bl) to -9.7 mV and significantly increased R(pc) from 391.6 kOmega to 2612.5 kOmega. Similar results were obtained with cyanide, but it remains unclear whether the large increases in R(pc) stem from the uncoupling of epithelial cells and/or the shutdown of conductive transport pathways in basolateral and apical membranes. Our results indicate that the apical membrane of principal cells is more than twice as conductive as the basolateral membrane. Partial ionic conductances suggest the rate-limiting step for transepithelial Na(+) secretion at the basolateral membrane. PMID- 12770113 TI - Energy metabolism and metabolic rate of the alder leaf beetle Agelastica alni (L.) (Coleoptera, Chrysomelidae) under aerobic and anaerobic conditions: a microcalorimetric study. AB - In early fall, adult alder leaf beetles (Agelastica alni L.) retreat, for overwintering, to the top layer of the soil near their forage trees where the ground gets easily waterlogged so that the beetles will be submerged and cut off from atmospheric oxygen. Hence, unlike most other adult insects, alder leaf beetles encounter hypoxia/anoxia in their natural habitat and this may occur at moderate temperature. Exposing beetles to pure nitrogen gas at 20 degrees C had similar behavioral and metabolic effects as submerging them in water, causing rapid immobility and increasing the content of lactate about sevenfold to some 5&mgr;molg(-1) body weight during 10h anoxia. Recovery from 10 h hypoxia/anoxia in pure nitrogen was complete within about 90min.Hypoxia/anoxia triggered a marked decrease in metabolic activity in the beetles (microcalorimetry at 21.7 degrees C) as indicated by a precipitous drop in their heat flow rate, from 1.39+/-0.27 to 0.08+/-0.04mWg(-1) body weight, i.e. by about 94%, when the flow of gas through the calorimeter was switched from air to pure nitrogen. Post anoxic recovery was accompanied by a peak in heat flow rate that exceeded the basal normoxic rate by about 50%. The homoeostasis of adenine nucleotides in Agelastica is lost when oxygen is wanting. Submergence at 15 degrees C for three days caused a dramatic fall in ATP, to less than 2% of the normoxic value, and a marked increase in AMP, while the total contents of adenine nucleotides decreased by almost two-thirds. Reduced metabolic activity, combined with the capacity to regenerate ATP after readmission of air, is regarded as a key factor for surviving transient lack of oxygen in alder leaf beetles. PMID- 12770114 TI - The potential for incorporation of male derived proteins into developing eggs in the leafhopper Bothrogonia ferruginea. AB - The leafhoppers, Bothrogonia ferruginea (Homoptera: Cicadellidae), eclose to adults in summer with immature reproductive organs. The adults live for 10 months including a hibernation of 4 months. Overwintered females mate multiply in spring. Eggs develop rapidly and are laid continuously in this mating period. Males produce sperm-bundles in which sperm are attached in a row to a rope-like hyaline material, and transfer them to the female via a large spermatophore that is placed in her bursa copulatrix. After mating, sperm are separated and removed to a spermatheca for storage prior to fertilization, but the sperm-binding material (trypsin degradable proteins) and the spermatophore disappear in the bursa and an enlarged portion of the genital duct. An injection of rhodamine B dyed proteins into the female bursa with a microsyringe results in the production of intensely fluorescent eggs developing in the ovaries. This suggests that females could incorporate proteinaceous material derived from male spermatophores and/or sperm-binding material into their oocytes. PMID- 12770115 TI - Tissue distribution and characterization of predominant hemolymph carrier proteins from Dermacentor variabilis and Ornithodoros parkeri. AB - The tissue distribution of the predominant hemolymph protein found throughout tick development was examined in the hard tick, Dermacentor variabilis, and in the soft tick, Ornithodoros parkeri. In D. variabilis, the predominant (purified) hemolymph protein was a lipoglycoheme-carrier protein (DvCP) with a molecular weight of 200K. A protein with a similar mobility on native-PAGE was found in fat body, salivary gland, muscle and ovary from partially fed females which was most abundant in the plasma and salivary gland. DvCP from plasma, salivary gland and fat body of partially fed females consisted of two subunits on SDS-PAGE (98 and 92K). In replete females, only salivary gland exhibited protein subunits equivalent to hemolymph CP. CP in salivary gland and fat body stained positive for lipids. The concentration of CP in tissues varied between partially fed and replete females, indicating a difference in the expression and/or sequestration of CP during adult development. The predominant hemolymph carrier protein from O. parkeri (OpCP) was purified to homogeneity for the first time and is presumed to have similar functions to CP from D. variabilis. Purified OpCP exhibited a molecular weight of 668K by native-PAGE. Unlike CP from D. variabilis, OpCP was not detected in fat body or salivary gland tissues but occurred abundantly in coxal fluid. By SDS-PAGE, purified hemolymph OpCP consisted of two major subunits (114 and 93K) and a less abundant protein with an apparent molecular weight of 48K. Purified native OpCP was a lipoprotein like DvCP. A spectral analysis of purified OpCP failed to demonstrate the presence of heme like that found for CP from D. variabilis, purified by the same methods. However, plasma from O. parkeri contained heme with a lambda(max) of 410nm. PMID- 12770116 TI - Lifetime ovipositional patterns of mated and virgin onion flies, Delia antiqua (Diptera: Anthomyiidae). AB - Over their 47.2+/-1.9 (mean+/-SEM) day lifespan, mated onion flies, Delia antiqua, oviposited more uniformly than did virgins. Mated females began ovipositing at 6.4+/-0.2 days old and regularly deposited 14.2+/-0.6 eggs/day for 3-4 weeks. Thereafter, oviposition slowed and stopped at about 1 week before death. Virgin flies began ovipositing 24.7+/-1.5 days into their 59.0+/-3.8 day lifespan, and deposited eggs at an increasing rate for 3-4 weeks, generating a mean overall ovipositional rate of 5.8+/-0.5 eggs/day. The later onset of virgin oviposition (ovipositional activation) and not a shorter ovipositional period largely accounted for the disparity between virgin and mated female lifespans. Mean lifetime egg production of mated females was 475+/-27 eggs versus 179+/-30 eggs for virgins. Ovipositional and post-ovipositional periods (34.4+/-1.8 and 7.2+/-1.0 days) for mated females were not significantly different from those of virgin females (30.9+/-3.6 and 6.7+/-1.2 days). Over 90% of virgin females laid eggs. The distinction between mated and virgin ovipositional patterns may be specifically attributed to differences in: (1) egg maturation, (2) age at ovipositional activation, and (3) egg-laying rate. PMID- 12770117 TI - Factors involved in the post-copulatory neural inhibition of pheromone production in Choristoneura fumiferana and C. rosaceana females. AB - Normal mating lasts approximately 3 h in Choristoneura fumiferana and C. rosaceana. Data generated from interrupted matings showed that the act of mating did not suppress pheromone production (pheromonostasis) in either species although, in C. rosaceana, pheromone titre declined slightly the night following mating. In both species the migration of sperm to the spermatheca (SP) occurred several hours after mating, and coincided with a significant and permanent depression in pheromone titre, as well as egg fertilisation and oviposition. However, disrupting matings within 2 h of the onset resulted in oviposition patterns similar to virgins in both species, with mostly infertile eggs being laid by C. fumiferana females while oviposition was totally inhibited in C. rosaceana. The transection of the ventral nerve cord (VNC) 1 h post-mating did not result in the depression of pheromone titres the following night in either species but if the VNC was transected 3 h post-mating, pheromonostasis was observed. While 25% of C. fumiferana females had sperm in their SP 2 h after mating, it took at least 4 h in C. rosaceana. This suggests that while the physical presence of sperm in the SP may play some role in the termination of pheromone production in C. fumiferana, other factors must trigger the neural signal that elicits pheromonostasis in both species. A better understanding of the temporal dynamics of both apyrene and eupyrene sperm within the different parts of the female reproductive system might clarify these interspecific differences. PMID- 12770118 TI - Acoustic sensitivity of fly antennae. AB - The vast majority of higher flies - the brachycerans - is endowed with strikingly similar antennal morphologies and elaborate antennal mechanosensory organs. Thus far, however, the function of audition has been attributed only to the antennae of Drosophilid and Tephritid flies. Antennal mechanical sensitivity to sound is now documented in a broad range of fly species. These results highlight the wide occurrence of audition in as many as 120,000 species of higher Diptera. Antenna based audition, as defined by the capacity of sensing the mechanical vibrations of the antenna in response to sound, thus appears to constitute an ubiquitous sensory capacity among higher flies. Functionally, antennal hearing can be used in the contexts of intraspecific acoustic signalling, flight control, and putatively close-range echolocation. PMID- 12770119 TI - Sperm survival in the female reproductive tract in the fly Scathophaga stercoraria (L.). AB - While sperm competition risk favours males transferring many sperm to secure fertilizations, females of a variety of species actively reduce sperm numbers reaching their reproductive tract, e.g. by extrusion or killing. Potential benefits of spermicide to females include nutritional gains, influence over sperm storage and paternity, and the elimination of sperm bearing somatic mutations that would lower zygote fitness.We investigated changes in sperm viability after in vivo and in vitro exposure to the female tract in the polyandrous fly, Scathophaga stercoraria. Sperm viability was significantly lower in the females' spermathecae immediately after mating than in the experimental males' testes. Males also varied significantly in the proportion of live sperm found in storage in vivo. However, the exact mechanism of sperm degradation remains to be clarified. In vitro exposure to extracts of the female reproductive tract, including female accessory glands, failed to significantly lower sperm viability compared to controls. These results are consistent either with postcopulatory sperm mortality in vivo depending entirely on the male (with individual differences in sperm viability, motility or longevity) or with postcopulatory sperm mortality being subtly affected by female effects which were not detected by the in vitro experimental conditions. Importantly, we found no evidence in support of the hypothesis that female accessory glands contribute to sexual conflict via spermicide. Therefore, female muscular control remains to date the only ascertained mechanism of female influence on sperm storage in this species. PMID- 12770120 TI - Midgut exopeptidase activities in Aedes aegypti are induced by blood feeding. AB - Midgut extracts from Aedes aegypti females exhibited hydrolytic activities against synthetic substrates for carboxypeptidase A, carboxyopeptidase B and leucine-aminopeptidase. The three activities showed a broad pH optimum, with maximum activities at pH between 6.5 and 8.5. Enzymatic activities were further characterized by testing the effects of a variety of protease inhibitors. Captopril and 1-10-phenantroline inhibited the activities of carboxypeptidases A and B, while leuhistin, amastatin and bestatin inhibited aminopeptidase activity. Exopeptidase activities were induced by a blood meal and the highest activities were found during the peak of trypsin activity, about 20-24h after feeding. An amino acid meal failed to induce significant increases in any of the three exopeptidase activities. The amounts of exopeptidase activities induced were proportional to the protein concentration of the meal. The addition of soy trypsin inhibitor to the protein meal blocked the post-feeding induction of exopeptidases. The features of the induction of synthesis of the three exopeptidase activities resembled the induction of synthesis of late trypsin during the second phase of digestion. PMID- 12770121 TI - Stylus of the odonate endophytic ovipositor: a mechanosensory organ controlling egg positioning. AB - Using light and scanning electron microscopy, a sensory field consisting of 15-20 campaniform sensillae is described on the base of the stylus of the endophytic ovipositor of Odonata. It is hypothesised that two symmetric styli equipped with this number of sensillae can function as a mechanosensory organ responsible for control of precise egg positioning in plant stems during oviposition. In laboratory experiments with females of damselflies Lestes sponsa and Lestes barbarus (Lestidae), it was demonstrated that the distance between laid eggs is not dependent on the presence of styli. Removal of styli from both sides did not influence a shift of oviposition to one side. Females with one removed stylus shifted the clutch line in the opposite direction toward the removed stylus. Additionally, removal of styli influenced positions of single eggs in egg sets, and disturbed the capacity for complex oviposition. Thus, both morphological and experimental data support the hypothesis that styli participate in the control of egg line and egg patterning in the clutch. PMID- 12770122 TI - Sensitive periods for wing development and precocious metamorphosis after precocene treatment of the brown planthopper, Nilaparvata lugens. AB - The critical periods for juvenile hormone suppression of wing development and metamorphosis were examined in a pure brachypterous line of Nilaparvata lugens following topical application of Precocene II (PII) to various stages of the third and fourth nymphal stadia. When PII, in doses ranging from 10 pg to 100 ng, was applied to 12-h-old third or 6-h-old fourth stadium nymphs, long-wing formation (macroptery) was induced. Macropter induction ranged from 5 to 50% in females and from 30 to 50% in males, the effect being more prominent in males. The sensitive period for macropter induction lasted from the early second stadium through to day one of the fourth (penultimate) stadium. Beyond that period nymphs required sensitivity to PII and the numbers of macropters increased towards the final nymphal molt, again males were more sensitive. On the other hand, periods sensitive to PII for precocious metamorphosis induction appeared from early second stadium to 18 h after molting to the third nymphal stadium. Overlapping application of JH-III with PII at early third stadium could totally inhibit the occurrence of precocious metamorphosis and significantly rescue the macropter induction. From the data described, a scheme is presented for the control of wing development and metamorphosis by juvenile hormone, assuming two types of threshold value determining wing form and metamorphosis. PMID- 12770123 TI - Temperature modulates epidermal cell size in Drosophila melanogaster. AB - Most ectotherms show increased body size at maturity when reared under colder temperatures. In principle, temperature could produce this outcome by influencing growth, proliferation and/or death of epidermal cells. Here we investigated the effects of rearing temperature on the cell size and cell number in the wing blade, the basitarsus of the leg and the cornea of the eye of Drosophila melanogaster from two populations at opposite ends of a South American latitudinal cline. We found that, in both strains of D. melanogaster and in both sexes, a decrease in rearing temperature increases the size of the wings, legs and eyes through an effect on epidermal cell size, with no significant change in cell number. Our results indicate that temperature has a consistent effect on cell size in the Drosophila epidermis and this may also apply to other cell types. In contrast, the evolutionary effects of temperature on the different organs are not consistent. We discuss our findings in the context of growth control in Drosophila. PMID- 12770124 TI - Physiological and biochemical aspects of flight metabolism in cocoon-enclosed adults of the fruit beetle, Pachnoda sinuata. AB - We studied several aspects of flight metabolism in cocoon-enclosed adults of the fruit beetle Pachnoda to investigate their flight capability. The majority of adults which were forcefully removed from their pupal cocoon flew off within 5 min of exposure to bright sunlight. Most of the beetles which did not fly voluntarily were, however, capable of flight. Compared with 2-4 week old adults of the same species, cocoon-enclosed adults have higher reserves of glycogen in flight muscles and fat body, whereas the level of total carbohydrates in the haemolymph and the concentration of proline in haemolymph, flight muscles and fat body were similar.Enzymes involved in carbohydrate breakdown (MDH, GAPDH) were more active in flight muscles and fat body of cocoon-enclosed adults compared with adults, while enzymes of proline metabolism in the flight muscles (AlaT, NAD ME) and fat body (AlaT, NADP-ME) had similar activities in cocoon-enclosed adults and adults. An enzyme of the beta-oxidation of fatty acids (HOAD) had similar activities in flight muscles and fat body of cocoon-enclosed adults and adults.Mitochondria isolated from flight muscles of adults removed prematurely from their cocoon favour the oxidation of proline and pyruvate. Pyruvate, however, is oxidized at higher rates than by mitochondria isolated from flight muscles of adults.During a short lift-generating flight, cocoon-enclosed adults proved that their flight muscles are capable of strong flight performance. During these flights, cocoon-enclosed adults consume proline and carbohydrates at a similar rate to that of adults.The endogenous AKH peptide, Mem-CC, has hyperprolinaemic and hypertrehalosaemic activity in cocoon-enclosed adults. The hypertrehalosaemic effect, however, is stronger in cocoon-enclosed adults than in adults.The content of Mem-CC in corpora cardiaca of larvae (3rd instar), cocoon enclosed adults and 1 day-old adults is similar at 5-6 pmol per pair of corpora cardiaca, whereas it is higher in 10 day-old adults and 20 day-old adults (37 and 15 pmol per pair corpora cardiaca, respectively).From these results we conclude that cocoon-enclosed adults comply with all the prerequisites for flight performance before they leave their pupal cocoon. Furthermore, cocoon-enclosed adults have a more pronounced carbohydrate-based metabolism before they leave their cocoon compared with adults, which suggests that carbohydrate breakdown is mainly involved in such activities as leaving the cocoon and burrowing activity thereafter. PMID- 12770125 TI - The propolis of stingless bees: terpenes from the tibia of three Frieseomelitta species. AB - The posterior tibia of foraging workers of three species of Frieseomelitta (Hymenoptera: Meliponinae) stingless bees have been shown to carry complex mixtures of plant-derived mono-, sesqui-, di- and tri-terpenes. These subtances were not found on the fore- or mid-legs, nor on other parts of the hind legs. F. silvestrii and F. silvestrii languida, when collecting, appear to exploit different plants for their resin even when housed in the same area. F. varia were found to be not collecting resin at the time of the initial sampling and were therefore sampled later. Mature foragers carry the resin. In the samples studied here, particularly prominent were the monoterpene alpha-pinene, the sesquiterpenes beta-caryophyllene, alpha-cubebene, alpha- and gamma-muurolene, gamma-cadinene, germacrene-D, and elemol and the diterpenes manool and totarol The collected material is used for the resin placed around the entrance to their nests and is also mixed with wax, to produce the cerum used for the structures in the nest. PMID- 12770126 TI - Motor control of the mandible closer muscle in ants. AB - Despite their simple design, ant mandible movements cover a wide range of forces, velocities and amplitudes. The mandible is controlled by the mandible closer muscle, which is composed of two functionally distinct subpopulations of muscle fiber types: fast fibers (short sarcomeres) and slow ones (long sarcomeres). The entire muscle is controlled by 10-12 motor neurons, 4-5 of which exclusively supply fast muscle fibers. Slow muscle fibers comprise a posterior and an antero lateral group, each of which is controlled by 1-2 motor neurons. In addition, 3-4 motor neurons control all muscle fibers together. Simultaneous recordings of muscle activity and mandible movement reveal that fast movements require rapid contractions of fast muscle fibers. Slow and subtle movements result from the activation of slow muscle fibers. Forceful movements are generated by simultaneous co-activation of all muscle fiber types. Retrograde tracing shows that most dendritic arborizations of the different sets of motor neurons share the same neuropil in the subesophageal ganglion. In addition, fast motor neurons and neurons supplying the lateral group of slow closer muscle fibers each invade specific parts of the neuropil that is not shared by the other motor neuron groups. Some bilateral overlap between the dendrites of left and right motor neurons exists, particularly in fast motor neurons. The results explain how a single muscle is able to control the different movement parameters required for the proper function of ant mandibles. PMID- 12770127 TI - Cellular signaling in eclosion hormone action. AB - Eclosion hormone (EH) is a 62 amino acid neuropeptide that plays an integral role in triggering ecdysis behavior at the end of each molt. At least three populations of cells are thought to be targets for EH, each of which show an EH stimulated increase in the intracellular messenger guanosine 3', 5' cyclic monophosphate (cGMP). These EH target cells are believed to include two pairs of neurons in each of the ganglia of the ventral nerve cord (VNC) that contain the neuropeptide crustacean cardioactive peptide (CCAP), the Inka cells of the peripheral epitracheal glands and intrinsic non-neuronal cells in the abdominal transverse nerves. This review describes likely signaling cascades that result in the EH-stimulated cGMP increase. Several lines of evidence suggest the involvement of a novel nitric oxide insensitive soluble guanylyl cyclase (GC). A novel GC with these properties has recently been identified and we also present evidence to suggest that it is activated by EH and describe possible pathways for its activation. In addition, we review our current knowledge on the cellular and molecular events that take place downstream of the increase in cGMP. PMID- 12770128 TI - Neural- and endocrine control of flight muscle degeneration in the adult cricket, Gryllus bimaculatus. AB - Neural- and endocrine mechanisms controlling degeneration of a dorsal longitudinal flight muscle, M112a, have been studied in adult Gryllus bimaculatus (Orthoptera: Gryllidae). Decapitation completely prevented muscle degeneration. Implantation of a pair of corpora allata or injection of juvenile hormone III into decapitated crickets caused muscle degeneration. Denervation of M112a resulted in reduction of muscle mass compared with that in sham-operated crickets. Denervation of M112a in decapitated crickets, however, did not affect muscle mass. Birefringence and ultrastructure of M112a showed an obvious regional difference in the onset of degeneration. Fibrillar structures of M112a always disappeared from the ventral to dorsal part. Distribution of axon terminals of motor neurons and mechanical responses to the motor nerve stimuli showed that M112a is composed of five motor units with similar twitch properties. When M112a was fully denervated, regional differences in degeneration disappeared. Partial denervation resulted in denervated muscle fibers losing birefringence earlier than innervated fibers. These results suggest that juvenile hormone causes breakdown of flight muscles, and neural factors control degeneration of flight muscles to some extent under the presence of the juvenile hormone. PMID- 12770129 TI - Developmental plasticity of the locomotor activity rhythm of Drosophila melanogaster. AB - We used four replicate outbred populations of Drosophila melanogaster to investigate whether the light regimes experienced during the pre-adult (larval and pupal) and early adult stages influence the free-running period (tau(DD)) of the circadian locomotor activity rhythm of adult flies. In a series of two experiments four different populations of flies were raised from egg to eclosion in constant light (LL), in light/dark (LD) 12:12h cycle, and in constant darkness (DD). In the first experiment the adult male and female flies were directly transferred into DD and their locomotor activity was monitored, while in the second experiment the locomotor activity of the emerging adult flies was first assayed in LD 12:12h for 15 days and then in DD for another 15 days. The tau(DD) of the locomotor activity rhythm of flies that were raised in all the three light regimes, LL, LD 12:12h and in DD was significantly different from each other. The tau(DD) of the locomotor activity rhythm of the flies, which were raised in DD during their pre-adult stages, was significantly shorter than that of flies that were raised as pre-adults in LL regime, which in turn was significantly shorter than that of flies raised in LD 12:12h regime. This pattern was consistent across both the experiments. The results of our experiments serve to emphasise the fact that in order to draw meaningful inferences about circadian rhythm parameters in insects, adequate attention should be paid to control and specify the environment in which pre-adult rearing takes place. The pattern of pre-adult and early adult light regime effects that we see differs from that previously observed in studies of mutant strains of D. melanogaster, and therefore, also points to the potential importance of inter-strain differences in the response of circadian organisation to external influences. PMID- 12770130 TI - Plant nitrogen status rapidly alters amino acid metabolism and excretion in Bemisia tabaci. AB - The effect of plant nitrogen (N) status on the content and distribution of free amino acids in the bodies and honeydew of silverleaf whiteflies Bemisia tabaci (Gennadius) Biotype B (= B. argentifolii Bellows and Perring) was determined. Whiteflies fed for 4 days on cotton leaves that received high or low N fertility. For low-N plants, photosynthesis and leaf total N levels were decreased, and a much-reduced amount of free amino acids was recovered in phloem sap. Low N fertility did not affect whitefly total N content, but did markedly decrease the free amino acid content. Glutamine, alanine and proline accounted for over half of the insect free amino acid pool for both N treatments. On a relative basis, adjustments in glutamine levels in response to plant N status were much larger compared to the other amino acids. Large amounts of amino N, especially asparagine, were excreted from whiteflies fed on high-N plants whereas amino N excretion essentially ceased for whiteflies fed on low N plants. The distribution of amino acids in the insects and honeydew was not closely related to the phloem sap amino acids. However, total amino acid excretion was quite indicative of the plant N status and the quality of the insect diet. The results indicated that whitefly free amino acid pools and excretion of amino N were rapidly altered by plant N status. PMID- 12770131 TI - Pharmacological characterisation of 5-hydroxytryptamine-induced contractile effects in the isolated gut of the lepidopteran caterpillar Spodoptera frugiperda. AB - The indolealkylamine 5-hydroxytryptamine (5-HT, 0.1nM-1&mgr;M) caused dose dependent increases in the number of contractions observed in guts isolated from the caterpillar Spodoptera frugiperda. Of the 5-HT analogues tested for agonist action, 2-methyl-5-HT (0.1-10&mgr;M) was a full agonist with reduced potency while alpha-methyl-5-HT (0.1-100&mgr;M), 5-carboxamidotryptamine (0.1-100&mgr;M), 5-methoxytryptamine (5-MeOT) (10nM-10&mgr;M), and tryptamine (1-100&mgr;M) were partial agonists. Incubation of isolated guts with proven mammalian 5-HT receptor antagonists showed that cyproheptadine (10nM-1&mgr;M), MDL 72222 (1-10&mgr;M), tropisetron (1-10&mgr;M) and 5-benzoyloxygramine (1-10&mgr;M) were potent non competitive antagonists of 5-HT-induced tissue contraction. In comparison, ketanserin (0.1-1&mgr;M) was a competitive antagonist. The mammalian selective serotonin reuptake inhibitors, clomipramine (10nM-10&mgr;M) and fluoxetine (10nM 10&mgr;M) also caused non-competitive inhibition of 5-HT-induced contraction while fluvoxamine (10nM-10&mgr;M) was a weak competitive antagonist. Low doses of clomipramine (0.1&mgr;M) caused potentiation of 5-HT-induced gut contraction thereby suggesting the presence of 5-HT reuptake systems in this tissue. The contractile effects of 5-HT were inhibited by verapamil, Li(+) and H7 and potentiated by theophylline thereby indicating that L-type Ca(2+) channels, phosphatidylinositol second messengers and cAMP, respectively, are involved in 5 HT-induced tissue contraction. The 5-HT receptors mediating contractility in the gut of S. frugiperda have properties in common with mammalian 5-HT(2) and Drosophila 5-HT(dro2A/2B) receptors. In addition, these data suggest that the tissue also contains receptors that are similar to mammalian 5-ht(6) and 5-HT(7) as well as Drosophila(dro1) receptors. However, the primary amino acid sequence of these lepidopteran 5-HT receptors will have to be elucidated before full comparisons can be made. PMID- 12770132 TI - The toxin Tx4(6-1) from the spider Phoneutria nigriventer slows down Na(+) current inactivation in insect CNS via binding to receptor site 3. AB - Tx4(6-1) a neurotoxic peptide from the venom of the aggressive South American 'armed' spider Phoneutria nigriventer, has been previously isolated and sequenced. It shows no detectable activity in mice but affects the peripheral nervous system of insects by stimulating glutamate release at the neuromuscular junction. Here we investigate possible interactions of the toxin with voltage activated sodium channels (Na(v)). We confirm that it is ineffective on mammalian Na(v) channels, and establish that it competes with the alpha-like toxin 125I-Bom IV, for binding on the site 3 of insect Na(v) channel (IC(50) value around 25nM). The physiological consequences of this binding to the insect Na(v) channel are shown by electrophysiology: Tx4(6-1) prolongs evoked axonal action potentials (APs) (<500&mgr;s duration in control). Prolonged 8-10ms or 'plateau' 500-800ms APs accompanied by repetitive firing at 80-150Hz are recorded after 4-8min of toxin action. This modification of evoked activity is due to a slowing down of sodium current inactivation. Effects of Tx4(6-1) on sodium current are compared with those of a typical scorpion alpha-toxin and of some other spider toxins active on insect Na(v) channels. At the end of long voltage pulses, the maintained inward sodium current may represent 50% of the peak current after scorpion alpha-toxin but only about 8-10% after spider toxins. To understand the slight differences in the effects of alpha-scorpion and spider toxins on the insect Na(v) channel, structural studies of toxin-channels interactions would be necessary. PMID- 12770133 TI - Timing and ecdysteroid regulation of the molt in last instar greenhouse whiteflies (Trialeurodes vaporariorum). AB - A system of markers has been devised to track the development of 3rd and 4th instar/pharate adult greenhouse whiteflies. Instars were identified based on measurements of body width and body length. Depending upon the host plant, the product of the two measurements was exceptionally useful in distinguishing between instars. Body depth was used to divide the 3rd instar into eight stages and body depth and color and appearance of the developing adult eye were used to divide the 4th instar/pharate adult into nine stages. Under conditions of L:D 16:8 and a temperature of 26+/-2 degrees C, the body depth of 3rd instars reared on greenbean increased from 0.025 (stage 1) to 0.2mm (stage 8) and the instar duration was approximately 3 days. The body depth of 4th instars increased from approximately 0.1+/-0.02 (Stage 1) to 0.3+/-0.03mm (Stage 5) and then remained constant or decreased slightly during adult development. Ecdysteroid titers peaked at approximately 120fg/&mgr;g protein during Stages 3 through 6 of the 4th instar. Based on an external examination of developing 4th instars and the fluctuations in ecdysteroid titer, it appears that adult development is initiated in Stage 4 or 5 4th instars. Results from histological studies support this view. In Stage 4 nymphs, a subtle change was observed in the corneagenous cells of the eye. However, most Stage 4 4th instars possessed wing development characteristic of earlier, immature stages. In all Stage 5 insects, wing development had been initiated and the corneagenous cells had become quite distinct. In Stage 6 whiteflies, the wing buds were deeply folded and by Stage 7, spines were observed on the new cuticle, indicating that the adult cuticle was well-formed by this stage. Our study is the first to investigate the timing and regulation of the molt, to monitor ecdysteroid titers in precisely staged 4th instar whiteflies and to examine the internal anatomical changes associated with metamorphosis in these tiny homopteran insects. PMID- 12770134 TI - Structure-activity relationship of contractile effects induced by helicokinins in the isolated gut of the lepidopteran caterpillar Spodoptera frugiperda. AB - The diuretic helicokinins YFSPWG-amide (Hez KI), VRFSPWG-amide (Hez KII) and KVKFSAWG-amide (Hez KIII) are potent contractants of the isolated gut of the caterpillar Spodoptera frugiperda at doses ranging from 0.1 to 10nM. In comparison, the pentapeptide FSPWG-amide was a full agonist with greatly reduced potency while SPWG-amide and PWG-amide were weak partial agonists. Substitution of individual amino acids in Hez KI with alanine revealed that replacement of the [phenylalanine(2)] residue caused a large fall in potency while replacement of [tryptophan(5)] residue caused complete loss of myogenic activity. The striking fall in potency of YASPWG-amide and the lack of activity of YFSPAG-amide confirm the requirement for aromatic groups in positions 2 and 3 of the core pentapeptide as well as supporting the ideas that the active core of these peptides adopts a beta-turn when interacting with receptors, bringing together the [Phe] and [Trp] residues that are critical for activity. Neither the pentapeptide proctolin nor the potent mammalian gut contractant Substance P were able to cause contraction when applied to caterpillar gut tissue. Incubation of isolated gut tissue in the phosphodiesterase inhibitor theophylline (10-100&mgr;M) caused significant potentiation of the response to applied Hez KI. Conversely, in the presence of the L-type Ca(2+) channel blocker verapamil (10&mgr;M-1mM) or Co(2+) (1-50mM) the contractile effects of Hez KI were attentuated significantly. These data suggest that the gut of S. frugiperda contains G-protein-linked kinin receptors that utilise cyclic AMP as their second messenger system and cause contraction by promoting the entry of extracellular Ca(2+). PMID- 12770135 TI - Comparison of the circadian eclosion rhythm between non-diapause and diapause pupae in the onion fly, Delia antiqua. AB - The influence of pupal diapause on adult eclosion rhythm of Delia antiqua was investigated. When non-diapause and diapause pupae were exposed to various photoperiods at 15, 20 and 25 degrees C, both of them emerged as adults close to the light-on time, but the phase of eclosion varied with photoperiod and temperature. Moreover, there was a significant difference in the eclosion time between non-diapause and diapause pupae; the eclosion peak of diapause pupae was earlier than that of non-diapause pupae. When non-diapause and diapause pupae were transferred to constant darkness (DD) after having experienced LD 12:12 at 15, 20 and 25 degrees C, both showed circadian rhythmicity in eclosion. Although the free-running period (tau) decreased slightly as temperature increased in both non-diapause and diapause pupae, the latter tended to show shorter tau than the former. This observation suggests that the observed difference in eclosion time in LD cycles between non-diapause and diapause pupae is due to differences in tau. PMID- 12770136 TI - Control of muscle degeneration following autotomy of a hindleg in the grasshopper, Barytettix humphreysii. AB - When the grasshopper, Barrytettix humphreysii, sheds a hindlimb during autotomy, certain thoracic muscles degenerate although they are neither directly damaged nor denervated. Muscle degeneration is induced when a leg nerve (N5) that does not innervate the thoracic muscles is severed. Together these results suggest that transneuronal mechanisms influence muscle survival. To further characterize this autotomy-induced process, we studied the degeneration of a thoracic tergotrochanteral muscle (M#133b,c) following autotomy or experimental manipulation in adult animals. Its degeneration is correlated with reduced activity of its neural input and occurs by programmed cell death (PCD). PCD onset is variable between individual muscle fibers, indicating that the trigger of degeneration is fiber specific. Muscle degeneration appears to be triggered by the loss of proprioceptive input from the autotomized limb, since severing of axons from proprioceptive organs, but not exteroceptive chemo- or mechanoreceptors, leads to muscle degeneration. Muscle disuse, neuronal degeneration, or changes in juvenile hormone titer do not appear to play a role in autotomy-induced degeneration. We propose that the loss of proprioceptive input from proximal campaniform sensilla on the tibia deafferents the thoracic muscle motor neurons and leads to a decrease in their activity. Muscle degeneration is ultimately triggered by the loss of normal neural activity. PMID- 12770137 TI - The role of endogenous antifreeze protein enhancers in the hemolymph thermal hysteresis activity of the beetle Dendroides canadensis. AB - Antifreeze proteins (AFPs) lower the freezing point of water by a non-colligative mechanism, but do not lower the melting point, therefore producing a difference between the freezing and melting points termed thermal hysteresis. Thermal hysteresis activity (THA) of AFPs from overwintering larvae of the beetle Dendroides canadensis is dependent upon AFP concentration and the presence of enhancers of THA which may be either other proteins or low molecular mass enhancers. The purpose of this study was to determine the relative contributions of endogenous enhancers in winter D. canadensis hemolymph.Winter hemolymph collected over four successive winters (1997-1998 to 2000-2001) was tested. The first three of these winters were the warmest on record in this area, while December of the final year was the coldest on record. Protein and low molecular mass enhancers raised hemolymph THA 60-97% and 35-55%, respectively, based on hemolymph with peak THA for each year collected over the four successive winters. However, the hemolymph AFPs were not maximally enhanced since addition of the potent enhancer citrate (at non-physiologically high levels) resulted in large increases in THA. (13)NMR showed that glycerol was the only low molecular mass solute present in sufficiently high concentrations in the hemolymph to function as an enhancer. Maximum THA appears to be approximately 8.5 degrees C. PMID- 12770138 TI - Secretion of beta-glycosidase by middle midgut cells and its recycling in the midgut of Tenebrio molitor larvae. AB - There are four beta-glycosidases (betagly1, betagly2, betagly3, and betagly4) in Tenebrio molitor midgut larvae. betagly1 and betagly2 have identical kinetic properties, and differ in a few amino acid residues. Purified betagly1 was used to raise antibodies in a rabbit. The resulting antiserum recognizes in a Western blot only betagly1 and betagly2 in midgut tissue homogenates and contents. An immunocytochemical study carried out using confocal fluorescence and immunogold techniques showed that betagly1+betagly2 are secreted by exocytosis mainly from the distal part of the second third of T. molitor midguts. This is the first immunocytochemical study of an insect digestive enzyme that does not have polymers as substrates. Enzyme assays with 0.3 mM amygdalin, a condition that detects only betagly1+betagly2, revealed that most of those beta-glycosidases are found in the lumen of anterior and middle midgut. This supports the hypothesis that a countercurrent flux of fluid occurs in T. molitor midgut that is able to carry betagly1 and betagly2 to anterior midgut, in agreement with the enzyme recycling mechanism thought to occur in most insects. PMID- 12770139 TI - Movement of spermatozoa in the reproductive tract of adult male Spodoptera litura: daily rhythm of sperm descent and the effect of light regime on male reproduction. AB - Sperm production and movement from the fused testes into the male reproductive tract of the common cutworm Spodoptera litura were studied in insects maintained in a 12h:12h light dark (LD) regime. Two types of sperm bundles, eupyrene (nucleated) and apyrene (anucleate) were present in the adult testes. Eupyrene bundles constituted about 25% of the total. Descent of spermatozoa from the testes into the upper vas deferens (UVD) first occurred about 24-30h before adult eclosion. On entering the reproductive tract, eupyrene spermatozoa remained in bundles while apyrene bundles became dissociated before they reached the UVD. Downward movement of both eupyrene and apyrene spermatozoa within the male tract occurred in a daily rhythm. Sperm descent from the testes into the UVD occurred during the early scotophase, followed by their further descent into the seminal vesicle (SV) during the photophase. Spermatozoa remained in the SV for only a short duration, whence sperm quickly passed through the lower vas deferens into the duplex, which acted as the main sperm storage organ until mating was initiated. During mating 80% of sperm left the duplex, but mating did not influence the number of sperm bundles that subsequently descended into the duplex or the rate of their descent. There was no evidence of sperm reflux. Rearing in constant light (LL) and in constant dark (DD) reduced the number of eupyrene sperm present in the testes of adults that emerged in LL and DD compared to controls (LD), although there was no significant effect on the number of apyrene sperm in the testes. The rhythmic pattern of sperm descent was suppressed in both LL and DD regimes, and the number of sperm in the duplex was adversely affected, with a marked impact in LL reared insects. Male longevity, mating behaviour, oviposition and fertility were found to be more severely affected in LL than in DD. PMID- 12770140 TI - Ultraviolet light sensitivity, unscheduled DNA synthesis and DNA repair in C7-10 Aedes albopictus mosquito cells. AB - We have examined the relative sensitivity of Aedes albopictus C7-10 mosquito cells to irradiation with ultraviolet light from a germicidal lamp. On the basis of plating efficiency, C7-10 cells were approximately two times more resistant to UV light than human 293 leukemia cells. Recovery after UV irradiation was accompanied by an increase in unscheduled DNA synthesis (UDS), which was measured by incorporation of (3)H-thymidine into acid-precipitable DNA in the presence of hydroxyurea. Under standardized conditions, UDS was maximal after a 10 min exposure (120 J/m(2)), and declined after longer exposures. In addition, UV treatment is associated with a small but reproducible increase in repair of plasmid DNA in transiently transfected cells. We anticipate that analysis of DNA repair activities in mosquito cells will identify molecular targets that might control longevity in transgenic mosquitoes. PMID- 12770141 TI - Patterns of selection: stress resistance and energy storage in density-dependent populations of Drosophila melanogaster. AB - Populations of Drosophila melanogaster subjected to extreme larval (CU) or adult (UC) densities for multiple generations were assayed for a variety of life history characters. When reared under either crowded or uncrowded larval conditions, populations which had been selected to tolerate the limitation of resources imposed by extreme larval (CU) crowding, exhibited greater starvation resistance and lipid content than did populations which do not routinely undergo larval density-dependent regulation. Previous studies have shown that the CU populations do not show a correlated increase in longevity; as has been generally observed for these characteristics in age-structured populations of D. melanogaster. This suggests that density-dependent natural selection may not always shape life histories of the same characteristic in the same direction that age-specific selection does. PMID- 12770142 TI - Effects of artificial diet containing GNA and GNA-expressing potatoes on the development of the aphid parasitoid Aphidius ervi Haliday (Hymenoptera: Aphidiidae). AB - Aphid parasitoids are important biological control agents. The possibility arises that whilst foraging on insect-resistant transgenic plants, they are themselves at risk from direct and indirect effects of the expression of a transgene used to control the pest species. A liquid artificial diet was successfully used to deliver the snowdrop lectin (Galanthus nivalis agglutinin; GNA) to the peach potato aphid, Myzus persicae. Bioassays utilising artificial diet incorporating GNA, and excised leaves of the GNA-expressing transgenic potato line, GNA2#28, were performed to assess the potential effects of GNA on the development of the aphid parasitoid Aphidius ervi. The results indicate that GNA delivered via artificial diet to the aphids can be transferred through the trophic levels and has a dose-dependent effect on parasitoid development. Parasitoid larvae excreted most of the ingested GNA in the meconium but some of it was detected in the pupae. Although A. ervi development was not affected when developing within hosts feeding on transgenic potato leaves, this probably reflected sub-optimal expression of the toxin in the transgenic potato line used PMID- 12770143 TI - Photoperiodic clock of diapause induction in Pseudopidorus fasciata (Lepidoptera: Zygaenidae). AB - Pseudopidorus fasciata enters diapause as fourth instar larvae at short day lengths. Using 24-h light-dark cycles, the photoperiodic response curves in this species appeared to be similar with a critical night length of 10.5h at temperatures below 30 degrees C. At an average temperature of 30.5 degrees C, the critical night length had shifted to between 15 and 17h. In experiments using non 24-h light-dark cycles, it was clearly demonstrated that the dark period (scotophase) was the decisive phase for a diapause determination. In night interruption experiments using 24-h light-dark cycles, a 1-h light pulse at LD12:12 completely reversed the long night effect and averted diapause in all treatments. At LD 9:15 light pulses of 1-h, 30- or 15-min also averted diapause effectively when both the pre-interruption (D(1)) or the post-interruption scotophases (D(2)) did not exceed the critical night length. If D(1) or D(2) exceeded the critical night length diapause was induced. The most crucial event for the photoperiodic time measurement in this species is the length of the scotophase. A 10-min light pulse placed in the most photosensitive phase reversed diapause in over 50% of the individuals. Night interruption experiments under non 24-h light-dark cycles indicated that the photoperiodic clock measured only D(1) regardless of the length of D(2), suggesting that the most inductive cycles are often those in which L+D are close to 24h. In resonance experiments, this species showed a circadian periodicity at temperatures of 24.5 or 26 degrees C, but not at 30.5 and 23.3 degrees C. On the other hand, Bunsow and skeleton photoperiod experiments failed to reveal the involvement of a circadian system in this photoperiodic clock. These results suggest the photoperiodic clock in this species is a long-night measuring hourglass and the circadian effect found in the final expression of the photoperiodic response in the resonance experiments may be caused by a disturbing effect of the circadian system in unnatural regimes. PMID- 12770144 TI - Revisiting water loss in insects: a large scale view. AB - Desiccation resistance in insects has long been thought to covary with environmental water availability, and to involve changes in both cuticular and respiratory transpiration. Here, we adopt a large-scale approach to address both issues. Water loss rate and precipitation are positively related at global scales. A significant proportion (68%) of the interspecific variation in water loss rate is explained at the genus level or above. The relationship between metabolic rate and water loss rate differs substantially between mesic and xeric species. While these variables covary as a consequence of their independent covariation with body mass in mesic species, this is not the case in xeric species. In the latter, there is a strong relationship between the residuals of the water loss rate-body mass and metabolic rate-body mass relationships, and water loss rate is much reduced. Moreover, because metabolic rate does not differ significantly between xeric and mesic species of a similar size, respiratory transpiration constitutes a greater proportion of total water loss in xeric than in mesic species of a similar size. This implies that respiratory transpiration and the extent to which it can be modified must be of considerable importance in xeric insect species, although finer scale studies suggest otherwise. PMID- 12770145 TI - The effects of Phaseolus vulgaris erythro- and leucoagglutinating isolectins (PHA E and PHA-L) delivered via artificial diet and transgenic plants on the growth and development of tomato moth (Lacanobia oleracea) larvae; lectin binding to gut glycoproteins in vitro and in vivo. AB - Red kidney bean, Phaseolus vulgaris, contains a lectin phytohemagglutinin (PHA) with toxicity towards higher animals. PHA exists in the isoforms PHA-E and PHA-L, which agglutinate erythrocytes and lymphocytes, respectively. Lacanobia oleracea larvae were reared from hatch on artificial diets containing PHA-E or PHA-L at 2% (w/w) dietary protein, and on transgenic Arabidopsis plants expressing either lectin at 0.4-0.6% of total soluble proteins. In artificial diet bioassays neither lectin affected larval survival, development, growth nor consumption. In transgenic plant bioassays both PHA-E and PHA-L promoted larval growth and development. This effect was greatest for PHA-E. Mean larval biomass of insects fed on plants expressing PHA-E was significantly greater (up to two-fold) than controls during the final two instars and the insects developed at a significantly greater rate so that after 26 days 83% of PHA-E exposed insects were in the final instar compared to 44% for control insects. PHA-E and PHA-L were detected by Western blotting in haemolymph, sampled from insects fed diets or plant material containing the lectins. However, despite the demonstrated potential for both isolectins to bind to gut glycopolypeptides in vitro neither was found to accumulate in vivo in the guts of exposed insects. Since lectin binding to gut polypeptides is thought to be necessary for insecticidal activity the failure of PHA-E and PHA-L to bind in vivo may account for their lack of toxicity to L. oleracea. PMID- 12770146 TI - Regulation of intracellular calcium in dispersed fat body trophocytes of the cockroach, Periplaneta americana, by hypertrehalosemic hormone. AB - Incubation of trophocytes from dissaggregated fat body of Periplaneta americana with either of the hypertrehalosemic hormones, HTH-I or HTH-II, leads to an increase in the cytosolic concentration of Ca(2+) from approximately 80 to approximately 310nM with a rise time of approximately 110s. The Ca(2+) concentration then declines to the resting level during the ensuing 5min. In the absence of extracellular Ca(2+) the increase in [Ca(2+)](i) due to HTH is limited to approximately 100nM. The calmodulin inhibitors calmidazolium and W-7 also limit to a similar degree the ability of HTH to increase [Ca(2+)](i). Phorbol 12 myristate 13-acetate, an activator of protein kinase C, was shown to block Ca(2+) entry through the plasma membrane. Additional evidence to support the view that HTH enhances Ca(2+) influx has been obtained by measuring the quenching of fura-2 fluorescence when Ca(2+) is replaced with Mn(2+). PMID- 12770147 TI - Eicosanoids mediate microaggregation reactions to bacterial challenge in isolated insect hemocyte preparations. AB - Nodule formation is the quantitatively predominant insect cellular defense reaction to bacterial challenges, responsible for clearing the largest proportion of infecting bacteria from circulation. It has been suggested that eicosanoids mediate several steps in the nodulation process, including formation of hemocyte microaggregates, an early step in the process. While fat body and hemocytes are competent to biosynthesize eicosanoids, the source of the nodulation-mediating eicosanoids remains unclear. To investigate this issue, we studied hemocyte microaggregation reactions to bacterial challenge in vitro. Hemocyte suspensions from the tobacco hornworm, Manduca sexta, were treated with the phospholipase A(2) inhibitor, dexamethasone, then challenged with the bacterium Serratia marcescens. Preparations treated with dexamethasone yielded fewer hemocyte microaggregations than untreated, control preparations. Furthermore, the influence of dexamethasone was reversed by amending experimental (dexamethasone treated) preparations with the eicosanoid biosynthesis precursor, arachidonic acid. Palmitic acid, which is not a substrate for eicosanoid biosynthesis, did not reverse the influence of dexamethasone on the microaggregation reaction. The influence of dexamethasone was also reversed by adding filtered media from challenged hemocyte preparations to dexamethasone-treated preparations. Finally, most hemocyte preparations treated with selected eicosanoid biosynthesis inhibitors formed fewer hemocyte microaggregations than control preparations. The 5- and 12-lipoxygenase inhibitor, esculetin, did not influence the formation of hemocyte microaggregations in this system. These results are consistent with similar investigations performed in vivo, and we infer that hemocytes are responsible for forming and secreting eicosanoids, which subsequently initiate nodulation by mediating hemocyte microaggregation. PMID- 12770148 TI - Quantification and development of teratocytes in novel-association host parasitoid combinations. AB - We studied the development of teratocytes derived from two congeneric gregarious endoparasitic species, Cotesia chilonis and C. flavipes, parasitizing two congeneric novel hosts, Diatraea saccharalis and D. grandiosella. The host parasitoid combinations studied allowed us to investigate relationships between host suitability and teratocyte development. D. saccharalis was a suitable host for both parasitoids, whereas D. grandiosella was suitable for C. chilonis development but often encapsulated C. flavipes progeny. Encapsulation of C. flavipes by D. grandiosella commenced around the time of parasitoid egg hatch, when teratocytes were released into the host's hemolymph. The gregarious parasitoids studied here released about 200 teratocytes per egg. Both absolute and normalized (teratocytes/parasitoid) numbers decreased over time. D. saccharalis supported more C. flavipes-derived teratocytes than D. grandiosella, possibly because of the unsuitability of the latter host. On intermediate assay days the number of C. flavipes-derived teratocytes was greater than for C. chilonis. However, C. chilonis-derived teratocytes grew larger than C. flavipes. Teratocytes in all host-parasitoid combinations doubled in size during parasitoid development. Teratocytes generally grew larger in D. grandiosella, which was a less suitable host. PMID- 12770149 TI - Water balance and osmoregulation in Stenocara gracilipes, a wax-blooming tenebrionid beetle from the Namib Desert. AB - Dehydration (10 days at 27 degrees C) of the Namib tenebrionid Stenocara gracilipes resulted in a rapid weight loss (17.5%), and a substantial decline in haemolymph volume (72%). Although the lipid content decreased significantly, metabolic water production was insufficient to maintain total body water (TBW). Rehydration (no food) resulted in increases in haemolymph volume, body weight (sub-normal), and TBW to normality. Haemolymph osmolality, sodium, potassium, chloride, amino acids, and sugars (trehalose and glucose), were all subject to osmoregulatory control during both dehydration and rehydration. Major osmolar effectors in this species are sodium, chloride, and amino acids, with most of the contribution to regulation of haemolymph osmolality coming from changes in the levels of these constituents. Changes in amino acid levels are not the result of interchange with soluble protein during dehydration (the possibility exists during extended rehydration, however). Despite faecal losses of sodium being low (8.2% of that removed from the haemolymph during dehydration), sodium concentrations do not return to normal during rehydration. Chloride concentrations increase supra-normally when access to water is allowed, and remain elevated throughout the rehydration period. Although faecal loss of potassium greatly exceeded the amount removed from the haemolymph (by approximately 1.8 times), haemolymph potassium levels were strongly regulated during rehydration. S. gracilipes demonstrates an exquisite capacity to regulate haemolymph osmolality under conditions of both acute water-shortage and abundance. Together with an efficient water economy (drinking when fog-water is available, and a superb water conservation mechanism in the form of wax-bloom production), this must serve to contribute to long-term survival of this species in an otherwise harsh abode. PMID- 12770150 TI - Infrared spectral sensitivity of Melanophila acuminata. AB - The spectral sensitivity of the pit organ of the beetle Melanophila acuminata (Coleoptera:Buprestidae) was measured using an ultrafast tunable infrared laser source and standard electrophysiological techniques. The pit organ may be classified as a broadband detector as the beetles responded to all infrared excitation wavelengths from 2 to 6&mgr;m. There was a decrease in response threshold and latency and an increase in the magnitude of the response in the region from 2.8 to 3.5&mgr;m, which corresponded to a region of decreased transmittance (increased absorbance) as measured by Fourier transform infrared spectroscopy. The implications of the correlation between spectral response and optical properties are discussed. PMID- 12770151 TI - Selective secretion of free glycine, a neutralizer against a plant defense chemical, in the digestive juice of the privet moth larvae. AB - The larva of the privet moth, Brahmaea wallichii (Brahmaeidae) is a specialist feeder of the privet tree, Ligustrum obtusifolium (Oleaceae). A very high concentration (50 mM or 0.4%) of free glycine, found in the digestive juice of the larvae, works as a neutralizer against the very strong protein-denaturing activity of privet leaves that is caused by oleuropein, an iridoid that functions in chemical defense. Concentration of free glycine was high in the anterior region of the midgut lumen and low in the posterior region. To examine if some glycine-specific secretion mechanism exists, injection experiments were performed using (15)N-labeled amino acids. When 13 &mgr;mol (1 mg) of (15)N-glycine was injected into hemolymph of fifth instar larvae of B. wallichii, a high concentration of (15)N (5 mM or 75 &mgr;g/g midgut content) was detected in the anterior parts of the midgut lumen 1 h after injection. (15)N-NMR data indicated at least 60% of the (15)N found in midgut lumen existed as (15)N-glycine. Approximately, 25% of the injected (15)N-glycine was estimated to have moved from the hemolymph to the midgut lumen. In contrast, no (15)N was detected in the midgut lumen when 13 &mgr;mol of (15)N-labeled alanine, lysine and glutamate were injected into hemolymph. Glycine was the only amino acid whose concentration was higher in the midgut lumen (50 mM) than in the hemolymph (22 mM). These data suggest the existence of some active and glycine-specific secretory mechanism in the midgut of B. wallichii. PMID- 12770152 TI - Feeding behaviour of morphologically similar Rhodnius species: influence of mechanical characteristics and salivary function. AB - Despite their morphological similarities, very similar Rhodnius species (R. prolixus, R. robustus, R. nasutus and R. neglectus) displayed a distinct feeding behaviour when fed on artificial feeder, pigeon or mouse. On pigeon hosts, these species showed distinct groups in terms of cumulative probing time - quicker species (R. prolixus and R. neglectus) followed by R. nasutus and finally a much slower species (R. robustus). On mouse hosts, R. nasutus showed quicker probing time compared to the other three species. Moreover, R. prolixus displayed quicker probing time compared to R. robustus and R. neglectus. Except for R. nasutus, the mean total ingestion rate tended to have different values between feeding sources (artificial feeder>pigeon>mouse). The volume ingested by each cibarial pump contraction and maximum frequency obtained using the artificial feeder are expected to be related to intrinsic mechanical characteristics of the insect feeding apparatus. However, probing time and the modulation of cibarial pump frequency on live hosts may be related to salivary function. R. prolixus showed high mechanical and salivary efficiency, achieving high values of total ingestion rate when fed on artificial feeder or either of the hosts. Comparative analysis suggests that species which possess higher total ingestion rates tend to achieve higher nutritional status, allowing them to reach higher densities. PMID- 12770153 TI - Age-specific mortality and reproduction respond to adult dietary restriction in Drosophila melanogaster. AB - Adult dietary yeast modulates mortality rate and reproduction of the Mediterranean fruit fly, Ceratatis capitata. In the medfly, a sugar-only diet leads to low mortality rates and reduced reproduction; addition of dietary yeast increases both mortality and egg laying. In Drosophila melanogaster low availability of dietary yeast is known to increase life span and reduce the rate of reproduction. Despite these similarities, because of differences in experimental design it remains unclear whether a common physiological mechanism modulates the effect of diet on survival. Here, we investigate how mortality rate and reproduction in D. melanogaster respond to the treatment regime used to study the medfly: no-yeast versus full diet. We find that adult medfly and D. melanogaster have opposite responses to the absence of yeast: D. melanogaster have high mortality when on no-yeast diet; when switched to full diet, D. melanogaster reduce mortality rates to the level presented by females continuously maintained on yeast. This reduction in mortality is accompanied by increased fecundity. These patterns are observed in all tested wildtype stocks, but flies made sterile by mutation in the gene oo18 RNA-binding protein (orb) lack this response. D. melanogaster, unlike medflies, appear to require adult dietary yeast to maintain maximal survival, and the capacity to assimilate yeast for somatic processes is one wildtype function of the gene orb. PMID- 12770154 TI - The physiology of locust phase polymorphism: an update. AB - The considerable progress made between 1990 and 1997 in locust phase-related research and in understanding the physiology of locust phase polymorphism is reviewed. The traits of locust phases are discussed and it is concluded that there are distinct strain-dependent differences in phase characteristics and their amplitudes even in the same species. Despite some advances, no major break through was achieved in the putative endocrine control of locust phase polymorphism. Phase-dependent differences in adipokinesis, flight fuels and migration of adult locusts, as well as novel methods in studying aggregation behaviour and activity of hoppers and adults, opened new lines in research of the physiology of locust phase polymorphism. Marked advances were made in phase related locust pheromone research, revealing, in Schistocerca gregaria, differences between the pheromonal system of the hoppers and that of the adults. These systems turned out to be more complex than previously assumed. Phenylacetonitrile, produced by sexually mature adult males, serving both as an attractant and a mutration-accelerating factor, was identified as the major compound of the adult pheromonal system in S. gregaria. A new aspect of transmission of phase characteristics from parent to progeny through the foam (froth) of the egg pod was revealed. Effects of some plant substances on locust phases were reported. However, no research has yet been published on the aspects of molecular biology of locust phase polymorphism. PMID- 12770155 TI - Feeding is not necessary for triggering plasticization of the abdominal cuticle in haematophagous bugs. AB - A simple device was used to quantify changes in the mechanical properties of the cuticle of Rhodnius prolixus and Triatoma infestans that take place when these insects feed, i.e., plasticization. Different stimuli were presented for 1min to test for their ability to trigger plasticization. These were: a blood meal, a Ringer solution meal, contact with a warm surface and thermal stimulation without such contact. Our results supplant any previous hypotheses that have supposed that the presence of food in the alimentary tract is necessary to evoke plasticization. We find that mere contact of the proboscis with a warm surface (without any food intake) is sufficient to trigger plasticization indistinguishable from that produced by a blood meal. Thermal stimulation alone, i.e., without physical contact, was not effective. PMID- 12770156 TI - Juvenile hormone effect on DNA synthesis and apoptosis in caste-specific differentiation of the larval honey bee (Apis mellifera L.) ovary. AB - Caste-specific differentiation of the honey bee ovary commences in the last larval instar. In this process, formation of germ cell clusters by synchronous and incomplete mitoses occurs in the queen ovary, whereas in the worker ovary programmed cell death is the dominant feature. BrdU and TUNEL labeling were used to study dynamics of cell proliferation and apoptosis-dependent DNA degradation in ovaries of naturally developing queens and workers, as well as in juvenile hormone-treated worker larvae. Cell proliferation in ovaries of last-instar queen larvae generally exceeded that in workers, except for the late feeding phase. This inversion in cell proliferation patterns coincided with the onset of apoptosis in worker ovaries, as evidenced by TUNEL labeling. Juvenile hormone application to early-fifth-instar worker larvae had two noticeable effects. First, it diminished the number of S-phase nuclei in ovaries of late feeding phase workers, bringing them to queen-like levels. Second, it prevented the induction of apoptotic DNA degradation. Caste-specific regulation of cell division in connection with programmed cell death can thus be attributed to the previously described differences in juvenile hormone titer in queen and worker larvae, adding a new facet to this hormone's multiple functions. PMID- 12770157 TI - The influence of brood on the pollen consumption of worker bees (Apis mellifera L.). AB - (1) In midgut dry weight (tissue plus contents) of worker bees we found a representative parameter for pollen consumption. Midguts of bees of successive ages were analyzed and correlated with various parameters. The relative proportions of sugar, protein and water were either constant or negatively correlated with midgut weight. Only the relative pollen weight (percent of midgut dry weight) increased. (2) To investigate the influence of different levels of brood on pollen consumption of individual bees, midgut dry weights from 2 normally breeding control colonies and 2 brood-reduced experimental colonies were analyzed. In bees from control colonies the pollen consumption increased up to the nursing age (3-10d), remained on an elevated level in middle-aged-bees (10 18d) and decreased relatively sharply towards the foraging ages (>21d). When queens were caged in the experimental colonies, the following decline of brood cells affected the consumption of pollen differently. After 6 days of caging, with a reduction of open brood only, no effect was seen. After 15 days, and even more pronounced after 23 days when no brood was present, the pollen consumption in young and middle-aged (10, 14, 18d) worker bees was significantly reduced, while it was clearly elevated in older bees. We discuss pollen consumption as an adaptation to reduced necessity to nurse brood in young and middle-aged bees, and to enhance life span in older animals. PMID- 12770158 TI - Insect larvae contain substances toxic to adults: the discovery of paralysins. AB - Acidic methanolic extracts of larvae of nine different insect species were found to contain substances that cause a lethal effect in the adult stage of the same species and of other species. These endogenous toxic substances, apparently being widely spread over the class of insects, were designated as paralysins, because of their immediate and observable paralytic effect upon injection. The developmental concentration curves of five different species of insects (Galleria mellonella (Lepidoptera), Neobellieria bullata (Diptera), Spodoptera frugiperda (Lepidoptera), Tenebrio molitor (Coleoptera) and Schistocerca gregaria (Orthoptera) indicate that the toxins are not present throughout all the developmental stages in the same concentration. The strongest paralytic activity was found in late instar larvae or in the early pupal stage. The temporal distribution of paralysins during development suggests that they might be involved in metamorphosis. PMID- 12770159 TI - 5-Methyl-2-phenyl-2-hexenal in the scent gland secretion of male cave crickets, Troglophilus cavicola and T. neglectus (Ensifera: Rhaphidophoridae). AB - 5-Methyl-2-phenyl-2-hexenal, a distinctly smelling compound, was identified as a constituent of the male scent gland secretion of two species of cave crickets, Troglophilus cavicola and T. neglectus. It is the first proof of a male specific exocrine compound in grasshoppers. A pheromonal function of the component could not be established; its possible biological role is discussed. PMID- 12770160 TI - Juvenile hormone and diapause in the Mediterranean corn borer, Sesamia nonagrioides. AB - Juvenile hormone content of Sesamia nonagrioides larvae reared under different environmental conditions that induce diapause was studied using a bioassay on newly-emerged Tribolium confusum pupae. Two analytical methods were also used to measure JHIII content. Extracts from larvae that developed under different conditions of photoperiod and temperature caused different effects in T. confusum pupae. This can be related to different diapause intensities. Extracts from diapausing larvae produced a higher juvenilizing effect than the expected, considering their JHIII titer calculated by the chromatographic analysis. This indicates that hormones other than JHIII must be present in extracts of diapausing larvae of S. nonagrioides. The analytical procedures confirm that diapausing larvae have a higher JHIII titer in the haemolymph than non-diapausing larvae. This shows that JH is involved in the maintenance of diapause in this species. PMID- 12770161 TI - Environmental hypoxia affects osmotic and ionic regulation in freshwater midge larvae. AB - The effect of anaerobic metabolism on the osmotic and ionic regulation of the extracellular fluid was examined. Larvae of three species, characterized by different hypoxia tolerance, were studied: Chaoborus crystallinus, Culex pipiens and Chironomus gr. plumosus. The use of the capillary electrophoresis technique made it possible to determine approximately 15 different ions from individual hemolymph samples. The hemolymph concentration of both inorganic and organic anions and cations as well as the osmolality were measured. A correlation between the hypoxia tolerance and the capability to avoid net changes in the ion concentration or in the osmolality of the three species studied here is proposed: Culex larvae, which have the lowest hypoxia tolerance, show a very large and very rapid lactate accumulation in their hemolymph under experimental hypoxia. This lactate accumulation is not compensated for by a change in the concentration of any other ion. Chaoborus larvae, with a medium hypoxia tolerance, utilize their very large hemolymph malate pool as a source of anaerobic energy. It is converted into succinate, thus inducing little net changes in the sum of the anions. There is a marked increase of the hemolymph osmolality, though. Chironomus larvae have the highest hypoxia tolerance and there are remarkably little changes in their hemolymph under hypoxia. Although these larvae are described as relying mainly on ethanol fermentation under environmental anaerobiosis, we demonstrated a marked lactate fermentation in severe hypoxia. The lactate accumulation observed in our study was compensated by a concomittant decrease of the hemolymph chloride concentration. PMID- 12770162 TI - Genetic variability in the area searched by a parasitic wasp: analysis from automatic video tracking of the walking path. AB - The ability of foraging hymenopterous parasitoid females to discover their hosts, and thus to be efficient agents in biological control programs, is likely to be related to the surface they are able to prospect per unit of time. However, this behavioural trait has never been accurately estimated, and its implication in female efficiency has never been really demonstrated. This paper provides an algorithmic method that can be used to estimate this trait from an automatic recording of the females' walking path. Using stochastic procedures simulating walking tracks, this trait is shown to be strongly related to the number of hosts that parasitoid females are able to attack per unit of time. This trait was estimated for individual females of Trichogramma brassicae Bezdenko (Hymenoptera; Trichogrammatidae). On average, females of this species are able to prospect about 28mm(2)s(-1). Finally, the genetic variation in this trait was studied using the iso-female line method. A significant genetic variability was observed. It provides the basic information that is necessary to start a genetic selection of mass-reared Trichogramma in order to improve their efficiency in controlling target pests in biological control programs. The functional and evolutionary implications of these results are discussed. PMID- 12770163 TI - Digestion of host immunoglobulin and activity of midgut proteases in the buffalo fly Haematobia irritans exigua. AB - The digestion of blood by the buffalo fly (Haematobia irritans exigua) was monitored for 6h at 33 degrees C after a single meal. Following the meal, the concentration of soluble protein within the midgut increased to a peak at 2 hours then decreased steadily over the next 4h. The magnitude of the increase in soluble protein at 2h indicated a release of protein from another source; most likely from lysed red blood cells. The immunoglobulin (IgG) fraction of the blood meal was digested rapidly (50% within one hour of feeding) and fully digested within 4h. This is indicative of its accessibility to digestive enzymes within the midgut. In contrast, when flies had continuous access to blood, the concentration of IgG in the midgut remained at a more constant level. The loss of antigen-binding activity of a specific antibody was more rapid than complete degradation of the IgG, with 70% of binding activity lost within one hour of feeding. The level of trypsin activity in the midgut increased from pre-feeding levels to reach a peak at 2h before returning to basal levels after 6h. The pattern of trypsin activity follows closely that of the concentration of soluble protein in the midgut (r=0.88). The activity of leucine aminopeptidase in the midgut also increased immediately after feeding and remained elevated for 4 h before declining to a basal level after 6h. The rapid digestion of IgG and subsequent loss of antibody activity suggests that for a specific anti-buffalo fly antibody to be effective it would need to be able to either evade the digestive system or induce a rapid response. PMID- 12770164 TI - A temporal profile of the endocrine control of trypsin synthesis in the yellow fever mosquito, Aedes aegypti. AB - Trypsin synthesis and secretion is induced after the female mosquito takes a blood meal. Its peak activity has been shown to be proportional to the amount and quality of food uptake. Further regulatory elements, hormones from the brain and the ovary, increase the synthethic rate of trypsin in the midgut by a factor of two. We investigated the temporal effect of removing the humoral factors by decapitation and ovariectomy. Trypsin synthesis was reduced to less than half its normal output when the operations were performed prior, or immediately after the blood meal. Postponing decapitation resulted in an increased activity. However, the dependence on hormones extended up to 14-16hrs after a meal, when maximal synthethic rates are assumed. Similarly, ovariectomy had a prolonged effect on trypsin synthesis. Finally, the lack of hormones reduced the synthetic capacity of the midgut even when small blood meals were given. We conclude that for continued efficient trypsin synthesis, humoral stimulation is necessary but is not part of the feedback mechanism that links the presence of food with the amount of trypsin secreted. PMID- 12770165 TI - The in vitro development from egg to prepupa of Campoletis sonorensis (Hymenoptera: Ichneumonidae) in an artificial medium: importance of physical factors. AB - A further attempt to improve the in vitro rearing of the solitary larval endoparasitoid Campoletis sonorensis (Hymenoptera: Ichneumonidae) was made after third instars were obtained in previous studies. Respiration was an important factor for the parasitoid's continued development: both small diet volumes that exposed the larval cuticle to air and O(2)greatly improved growth and development of C. sonorensis. Fifth instars were obtained when the parasitoid larvae were reared in medium volumes of 0.25ml and 0.1ml per well. Increasing the O(2) tension of the air resulted in larger larvae and prepupae. The agar layer under the nutritional medium was designed to slowly remove water from the diet. The results indicated that a drier environment was important for the parasitoid at the terminal stage. One percent and 0.8% agar gels, which removed water from the diet more rapidly, provided a drier environment and resulted in improved larval growth compared to the 0.6% agar gel. The detrimental effect of higher concentration of chicken egg yolk mixed in the diet was eliminated by using a dry egg yolk layer covered by an agar layer. This sandwich technique resulted in more parasitoid larvae molting to the final larval instar (5th) and the formation of many prepupae. However, the development of prepupae was premature and none of the fifth instar larvae successfully formed a normal cocoon. PMID- 12770166 TI - Behavioral and electrophysiological dose-response relationships in adult western corn rootworm (Diabrotica virgifera virgifera LeConte) for host pollen amino acids. AB - A strong correlation is shown between taste cell inputs and phagostimulatory outputs with predominant dietary pollen amino acids for western corn rootworm, Diabrotica virgifera virgifera. Behavioral and electrophysiological dose-response profiles in adult beetles are presented for five major free amino acids in host pollens. Differential responses were found with strongest phagostimulation and sensory response elicited by L-alanine and L-serine, followed in order by L proline and beta-alanine. gamma-Aminobutyric acid gave the weakest and most sporadic response. ED(50) values for phagostimulation and chemosensory input were 28.3nmol/disk and 13mM, respectively, for L-alanine and 17nmol/disk and 11mM, respectively, for serine. Threshold values for the responses were approximately 1 2mM. These behavioral and chemosensory dose-response ranges correspond closely to levels of free amino acids present in host plant pollens. Use of these response values in development of a pollen chemosensory code for western corn rootworm feeding is discussed. PMID- 12770167 TI - Sugar response differences related to sensillum type and location on the labella of Protophormia terraenovae: a contribution to spatial representation of the stimulus. AB - The chemoreceptor spike activity in response to sucrose in the concentration range 1-500mM was recorded from each of the 11 Intermediate and 13 Large labellar sensilla in the blowfly Protophormia terraenovae. The results showed that: (1) three of the four cells present in each sensillum are activated by sucrose stimulation; (2) differences between the Large and Intermediate types exist in the dose-response profiles of one of these cells (the 'water' cell), possibly reflecting different sugar receptor site populations on the dendritic membranes of homologous cells in the two types; (3) sensilla of both types are differentially responsive to sucrose solutions according to their location on the labellum. These differences may provide elements for a spatial representation of the stimulus source within the sensory coding process. PMID- 12770168 TI - Electroantennogram responses of Douglas-fir seed chalcids to plant volatiles. AB - Douglas fir seed chalcids females oviposit specifically into Douglas cones which they locate using olfactory and visual cues. We have tested whether this specialization was correlated with a specialized sensitivity towards cone volatiles. Field collected adults were presented a series of pure volatile chemicals. Electroantennogram responses (EAG) were recorded to generally occurring terpenoids and straight chain alcohols and aldehydes found in flowers and plant leaves. Monoterpenes identified in Douglas cone headspaces (alpha pinene, beta-pinene, 3-thujene, alpha- and beta-phellandrene, gamma-terpinene and myrcene) elicited lower EAG responses than fatty acid derivatives corresponding to green odours. The EAG response profile of females differed significantly from that of males. Females were very sensitive to hexanol-1, heptanal, (Z)-3 hexenol 1, terpineol and terpinen-4-ol. Male responses were significantly higher to humid air, nerolidol, thujan-4-ol, hexanal, carvacrol, piperitone and farnesol. Several compounds (Z- verbenol, carvone, jasminol, geraniol, nerol and eugenol) elicited long lasting electrophysiological responses (over 5 s) in both sexes. PMID- 12770169 TI - Action of estradiol-17beta on the synthetic activity of the silk gland in Bombyx mori L. AB - The effects of estradiol-17beta (E(2)) were studied on several metabolic parameters in the silk gland of Bombyx mori L. race Nistari. Topical application of different doses (0.05-4.0&mgr;g/g body weight) of E(2) on the first and second day of the fifth instar larvae showed a dose dependent effect when studied on the fifth day. A significant increase in silk gland weight and fibroin content was observed between the doses 0.05 and 0.1, and 0.1 and 1.0&mgr;g/g of E(2). A similar pattern of dose-dependent rise in DNA and RNA content of posterior silk gland (PSG) was observed with the doses of E(2) when the contents were expressed per pair of PSG. Higher doses of E(2) (2.0 or 4.0&mgr;g/g) demonstrated relatively less increase, unchanged level or a decrease in the above parameters in comparison to the control values. The glutamate-pyruvate transaminase of PSG showed a significant increase from 0.1 to 2.0&mgr;g/g of E(2) doses in comparison to the control value. Simultaneous injection of ICI-182780 (1.0&mgr;g/g), a very pure and specific antiestrogenic compound, with E(2) (1.0&mgr;g/g) caused a significant counteraction of E(2)-induced increase in silk gland activity, which was reflected in DNA and RNA content of PSG, wet weight and fibroin content of silk gland, and on glutamate-pyruvate transaminase activity. Cycloheximide (0.5&mgr;g/g), a protein synthesis blocker, caused a significant inhibition of the E(2) (1.0&mgr;g/g)-induced silk gland activity when treated along with estradiol. From this study it appears that estradiol has a specific effect on silk gland function and that it may act in a nuclear mediated way. PMID- 12770170 TI - The Apis mellifera pupal melanization program is affected by treatment with a juvenile hormone analogue. AB - Apis mellifera treated during different developmental phases with pyriproxyfen, a juvenile hormone analogue, show profound alterations in cuticular pigmentation and sclerotization. When the treatment is effected during the feeding phase of the fifth larval instar (LF5), the pupal development is blocked and pigmentation does not occur. Treatment of older larvae, at the spinning phase of the fifth larval instar (LS5), of prepupae (PP) or pupae at the beginning of the pupal period (Pw, white-eyed, unpigmented cuticle pupae) does not impair pigmentation, but, instead, this process is accelerated, intensified and abnormal. Hormonal treatment during these developmental phases (LS5, PP and Pw) induces earlier activity of phenoloxidase, an enzyme of the reaction chain leading to melanin synthesis. Treated pupae have significantly higher enzymatic levels and show a graded response in phenoloxidase activity after treatment with 0.1, 1 or 5&mgr;g pyriproxyfen. Besides pigmentation, other developmental events were also altered in treated bees: pupal development was shortened, and the expression of esterase 6 activity, the onset of which coincides with the beginning of pigmentation, was shifted with the precocious initiation of this process in treated pupae. The significance of these results is discussed in relation to the mode of hormonal action on cuticular pigmentation in insects. PMID- 12770171 TI - Synthesis of the same two proteins prior to larval diapause and pupation in the spruce budworm, Choristoneura fumiferana. AB - Spruce budworm larvae produce large quantities of two proteins (Choristoneura fumiferana diapause associated proteins 1 and 2, CfDAP1 and CfDAP2) that are diapause related. These proteins appeared soon after hatching and increased in abundance, reaching maximum levels by four days into the 1st instar, and they remained at high levels until three days after the termination of diapause. These two proteins were purified to homogeneity and their NH2-terminal sequences were obtained. Oligonucleotide primers designed on the basis of these NH2-terminal sequences were used in RT-PCR to isolate the cDNA fragments coding for these proteins. These PCR fragments were then used as probes to isolate the cDNAs that contained the complete coding region. The 2.5kb mRNAs coding for these proteins started to appear 24hr after hatching and large quantities of these mRNAs were detected in 1st instar and 2nd instar larvae until the 2nd instar larvae entered diapause. Low levels of these mRNAs were detected in the 2nd instar larvae that were preparing to enter diapause, in those that were in diapause as well as in those that terminated diapause. Low levels of CfDAP1 mRNA were also detected on days 1 and 2 after ecdysis to the 3rd instar. However, no CfDAP1 and CfDAP2 mRNAs could be detected during the 4th and 5th instar larval stages. The mRNAs reappeared 24hr after the 5th instar larvae molted into the 6th instar and increased to reach maximum levels by 60hr after ecdysis. The mRNA levels remained high until 156hr after ecdysis into the 6th instar (36-48hr before pupal ecdysis), after which they disappeared once again. Immunocytochemical analyses showed that CfDAP1 protein was present in 2nd and 6th instar larval fat body but not in 5th instar larval fat body. Thus, the same two genes were expressed for the first time before C. fumiferana larvae entered diapause and for a 2nd time before pupation. PMID- 12770172 TI - Ovarian development induced in decapitated female Culex pipiens pallens mosquitoes by infusion of physiological quantities of 20-hydroxyecdysone together with amino acids. AB - Infusion of 20-hydroxyecdysone into the hemocoel of unfed decapitated female Culex pipiens pallens mosquitoes, at a very low rate of 500-2000pg per day, often stimulated oogenesis of this species, when the hormone was infused together with amino acids. The hormone alone or amino acids alone showed no such stimulatory effect. Previous reports that using an abdomen ligated immediately after a blood meal for hormone injection reduced the quantity of 20-hydroxyecdysone needed to activate unfed female Aedes aegypti by a few thousand times, are therefore due mainly to a sufficient supply of amino acids from the midgut in the isolated abdomen. PMID- 12770173 TI - The radiochemical assay for juvenile hormone biosynthesis in insects: problems and solutions. AB - Measurement of juvenile hormone (JH) production using the radiochemical assay (RCA) for JH biosynthesis and release is usually a reliable and precise technique. However problems with radiolabeled precursors and misunderstanding of the data, the techniques and the calculations have contributed towards uncertainty with respect to published experimental results. Problems with the purity of [methyl-3H]-methionine or determination of its specific radioactivity have had detrimental effects on the reliability of results using the RCA. Proper control procedures and the use of 14C/3H-double-label RCA can be useful in detecting irregularities in the experimental results, and in determining contributing factors to any problems. The use of [methyl-14C]-methionine and an awareness of normally expected RCA values can also assist the researcher in checking the validity of results. The radiolabeled methyl moiety of methionine is incorporated into JH without discrimination relative to unlabeled methyl methionine, by the o-methyl transferase. However unexpected preferential incorporation of the [methyl-14C]- vs. [methyl-3H]-moiety into JH occurs, but is only evident at concentrations of radiolabeled methionine outside the normal range of the RCA. Changes in radioactive precursor formulation have no effect on the RCA. PMID- 12770174 TI - Proteolytic breakdown of the Neb-trypsin modulating oostatic factor (Neb-TMOF) in the hemolymph of different insects and its gut epithelial transport. AB - The degradation of the unblocked hexapeptide, trypsin modulating oostatic factor of the flesh fly Neobellieria (Sarcophaga) bullata (Neb-TMOF) was studied in vitro in the hemolymph of the lepidopteran Spodoptera frugiperda, the orthopteran Schistocerca gregaria and the dictyopteran Leucophaea maderae. The half-life in the different species varied from approximately 3min in L. maderae to approximately 25min in S. gregaria. Purification of the degradation products and ESI-Qq-oa-Tof mass spectrometry revealed the fragments Asn-Pro-Thr-Asn, Leu-His and Asn-Pro, which were the same in the hemolymph of all species. Except in Leucophaea, Neb-TMOF was cleaved in dipeptides starting from the C-terminus and the reaction could be, at least partially, inhibited by captopril. These observations suggest that a dipeptidase, which has very similar enzymatic properties as mammalian angiotensin converting enzyme (ACE) and which circulates in the hemolymph, apparently is involved in the breakdown of Neb-TMOF and might be a common but not a universal enzyme in insect hemolymph.The introduction of Neb-TMOF into the gut of S. gregaria with the help of a capillary tube (intubation) demonstrated that the intact peptide is able to cross the gut epithelium and to appear in the hemolymph compartment. Since [3H]-inulin, which is too large to cross cell membranes, was found to penetrate the gut walls at a measurable rate, the paracellular pathway might be also permeable to smaller peptides. There was indeed a clear correlation between the molecular weight of inulin, Neb-TMOF, and inositol and the rate of penetration of these compounds through the gut epithelium to the hemolymph. These are promising findings in view of a potential use of such peptides for insect control purposes. PMID- 12770175 TI - Juvenile hormone and aggression in honey bees. AB - We determined whether defense by individual bees against non-nestmates in honey bees (Apis mellifera) is correlated with their juvenile hormone (JH) titers, which are known to vary developmentally and seasonally. We bioassayed winter and summer bees for aggressive and non-aggressive individuals. Bees in winter could not be distinguished by task group, but bees in summer were segregated into nurses and guards. JH titers were correlated with aggressive behavior at two levels. First, winter bees and summer nurses, known to have lower JH titers, both showed less aggression toward foreign bees than did summer guards. Second, aggressive individuals had significantly higher JH titers than did non-aggressive bees within each colony. Inter-colonial variation in aggressiveness was maintained during summer and winter, suggesting a genetic basis for these differences. An alarm pheromone test further substantiated the existence of inter colonial differences. We found significant variation in JH titers among different colonies, but this variation was not significantly associated with colony-level aggressiveness. The correlation between JH and levels of aggressiveness within a colony suggests a regulatory role for JH, but variation among colonies involves factors other than JH. PMID- 12770176 TI - Effects of juvenile hormones and precocenes on the immune response of Spodoptera littoralis larvae to supernumerary larvae of the solitary parasitoid, Microplitis rufiventris Kok. AB - The study represents one of the first direct tests of juvenile hormones (JHs) on the cellular immune response of a lepidopteran insect, Spodoptera littoralis (Boisd.), superparasitized by a solitary parasitoid, Microplitis rufiventris Kok. JHII-treated S. littoralis larvae exhibited a significant reduction in encapsulation response to the supernumerary M. rufiventris larvae at different developmental ages (0-4, 10-14 and 24-26 h) of supernumerary M. rufiventris embryos within 48 or 144 h after parasitization than either JHI-treated or control hosts. The reduction in encapsulation response was significantly greater in hosts which received higher JHII dose (5 &mgr;g) than those treated with lower one (1 &mgr;g).The progression of the encapsulation, culminating in melanization in the capsules occurred more frequently in precocene-treated hosts than either JH-treatment or control ones. Application of the anti-juvenile hormones (PI or PII) caused stronger encapsulation reaction to the injured supernumerary parasitoid larvae than JHI or JHII. In PI-treatments, the reaction was rapid and dramatic by lower (25 &mgr;g) dose treatments than higher ones (70 &mgr;g).The capsules formed around the supernumerary larvae of M. rufiventris showed signs of melanization within 48 h in PI-treated hosts. Melanization was more pronounced in either PI- or PII-treated hosts prior emergence of successful parasitoid larvae. Generally, the injured supernumerary larvae were seen within melanin-rich capsules in PI- and PII-treated hosts. Melanization was not observed in most capsules of JHI- and JHII-treated hosts. Additional physiological effects were observed: (1) in some cases, application of JHI to parasitized S. littoralis larvae inhibited subsequent emergence of its wasp, M. rufiventris, and (2) teratocytes (cells of wasp origin) from PI or PII-treated hosts were smaller in size than those observed in the JH-treated or control hosts.Finally, the dramatic increase in encapsulation responses of precocene-treated S. littoralis larvae of the supernumerary of M. rufiventris and definitely opposite reactions in typical hosts but JHII-treated hosts suggested that the cellular defense reaction may be under inhibitory hormonal control.The results of the study have implications for medically important insects such as mosquitoes and their interactions with parasites, as well as having significance for biocontrol and mass rearing programs using parasitoids. PMID- 12770177 TI - The relationship between 'critical weight' and 20-hydroxyecdysone in the female ixodid tick, Amblyomma hebraeum. AB - THE FEEDING CYCLE OF ADULT FEMALE TICKS (ACARI: Ixodidae) is divided into preparatory, slow and rapid feeding phases. At the transition from slow to rapid feeding, Amblyomma hebraeum (Koch) females reach a 'critical weight' (CW; approx. 10x the unfed weight) that is characterized by several behavioural and physiological changes. Five of these changes were used as criteria to establish a more precise estimate of CW than we have to date. The CW as defined by re attachment to the host was 9x the unfed weight, while for haemolymph ecdysteroid titre, salivary gland degeneration, ovary weight, oocyte length and oocyte vitellin content the CW was 10x, 10x, 12x, 12x and 13x, respectively. CW thus varies depending on the parameter measured. Although previous studies have established the influence of ecdysteroids on salivary gland degeneration and vitellogenesis, here we demonstrate a further effect of ecdysteroids: inhibiting re-attachment to the host. PMID- 12770178 TI - Malfunction of circadian clock in the non-photoperiodic-diapause mutants of the drosophilid fly, Chymomyza costata. AB - The diel rhythmicity of adult eclosion was recorded in reciprocal F1 hybrids between the wild-type (Sapporo) and mutant (NPD) strains of Chymomyza costata and the functionality of central circadian clocks was checked in both strains by assessing diel and circadian patterns of the per gene mRNA abundance oscillations in fly heads using competitive polymerase chain reaction methodology. The previously detected mutations in the per coding region of the NPD strain (Shimada, Entomol. Sci. 2 (1999) 575) were found to be primarily neither responsible for the loss of the eclosion rhythm nor for the malfunction of the circadian clocks. While distinct diel and circadian rhythms in per mRNA abundance were found in the wild-type flies, the npd-mutants showed constant (arrhythmic) and low abundance of the per mRNA transcripts. Because the non-photoperiodism, arrhythmicity of adult eclosion and the malfunction of central circadian clocks all seem to result from a mutation in the autosomal npd locus, we hypothesize, that a product coded by this locus may represent a 'point of contact' between the circadian and photoperiodic time measurement systems in C. costata. PMID- 12770179 TI - Hormone-dependent protein patterns in integument and cuticular pigmentation in Apis mellifera during pharate adult development. AB - The epidermal proteins from staged Apis mellifera pupae and pharate adults and the progress of cuticular pigmentation until adult eclosion were used as parameters to study integument differentiation under hormonal treatment. Groups of bees were treated at the beginning of the pupal stage with the juvenile hormone analog pyriproxyfen (PPN) or as pharate adults with 20-hydroxyecdysone (20E). Another group was treated with both hormones applied successively at these same developmental periods. Controls were maintained without treatment. The epidermal proteins, separated by SDS-PAGE and identified by silver staining, were studied at seven intervals during the pupal and pharate adult stages. The initiation and progress of cuticular pigmentation was also monitored and compared to controls. The results showed that PPN reduced the interval of expression of some epidermal proteins, whereas 20E had an antagonistic effect, promoting a prolongation in the time of expression of the same proteins. In PPN-treated bees, cuticular pigmentation started precociously, whereas in 20E-treated individuals this developmental event was postponed. The double hormonal treatment restored the normal progress of cuticular pigmentation and, to a large extent, the temporal epidermal protein pattern. These results are discussed in relation to the 20E titer modulation and morphogenetic hormone interaction. PMID- 12770180 TI - Induction of digestive alpha-amylases in larvae of Zabrotes subfasciatus (Coleoptera: Bruchidae) in response to ingestion of common bean alpha-amylase inhibitor 1. AB - Zabrotes subfasciatus larvae possess three alpha-amylase isoforms determined by in gel assays following SDS-PAGE. Two minor isoforms present lower electrophoretic mobility than the major form. When developed inside Vigna unguiculata (cowpea) seeds, fourth instar larvae have minor quantities of the slow-migrating isoforms, but when reared on seeds of Phaseolus vulgaris (common bean), the two slow-migrating forms are expressed in higher amounts, whilst the quantity of the major constitutive form is independent of the host bean. Larvae at the beginning of the fourth instar were fed on flour or cotyledons of cowpea and common bean and it was observed that the larvae fed on the common bean expressed the two slow-migrating forms in higher amounts when compared to the control larvae fed on cowpea. In order to investigate the possible correlation between the induction of alpha-amylases and the ingestion of the common bean alpha-amylase inhibitor 1 (alphaAI-1), this inhibitor was incorporated into artificial diet. It was observed that larvae fed on diet containing chronic doses of alphaAI-1 during their development, produced the two slow-migrating forms in higher amounts than control larvae, however, fourth-instar larvae fed on the same diet presented less amylase activity than control larvae. The data suggested that alphaAI-1 is involved in amylase induction and that it has inhibitory activity against the constitutive amylase, when starch granules are used as substrate. PMID- 12770181 TI - Physiological response of Colorado potato beetle and beet armyworm larvae to depletion of wound-inducible proteinase inhibitors in transgenic potato plants. AB - Larvae of Colorado potato beetle (CPB), Leptinotarsa decemlineata, and beet armyworm (BAW), Spodoptera exigua, reared on potato plants in which wound-induced accumulation of proteinase inhibitors (PIs) was largely reduced through antisense mediated depletion of a specific lipoxygenase (LOX H3) had significantly larger weight gains than those fed on non-transformed plants. The midgut endoproteolytic activities of CPB larvae fed on non-transformed potato were significantly higher than those from larvae fed on LOX-H3-deficient plants. However, none of these proteolytic activities was inhibited by potato leaf extracts, regardless of the plant that they were fed on. Taken together, these data suggest that CPB, a leaf feeding specialist of solanaceous plants, is largely adapted to the inducible PIs of potato, though the metabolic cost associated with the hyperproduction of digestive proteases may account for the 14-31% lower weight gain of larvae fed on non-transformed plants. The effect of LOX-H3 depletion on insect performance was more evident with larvae of the polyphagous BAW (52-63% higher weight gain and 73% higher fecundity when reared on LOX-H3-deficient plants). The poorer larval performance of BAW on non-transformed plants may be due to the susceptibility to inhibition by potato leaf tissues of most BAW digestive proteases. Indeed, BAW larvae fed on non-transformed potato showed a significant reduction in most endoproteolytic activities compared to larvae fed on LOX-H3-deficient plants, suggesting a that these insects deal poorly with induced plant defences in potato. PMID- 12770182 TI - Larval crowding in Drosophila melanogaster induces Hsp70 expression, and leads to increased adult longevity and adult thermal stress resistance. AB - In this study we show for the first time that moderate high larval density induces Hsp70 expression in Drosophila melanogaster larvae. Larval crowding led to both increased mean and maximal longevity in adults of both sexes. Two different measures of heat-stress resistance increased in adult flies developed at high density compared to flies developed at low density. The hardening-like effect of high larval density carried over to the adult life stage. The hardening memory (the period of increased resistance after hardening) was long compared to hardening of adult flies, and possibly lasts throughout life. The increase in resistance in adults following development at high larval density seemed not to be connected to Hsp70 itself, since Hsp70 expression level in adult flies after hardening was independent of whether larvae developed at low or high densities. More likely, Hsp70 may be one of many components of the stress response resulting in hardening. PMID- 12770183 TI - Effects of aggression and wing removal on brain serotonin levels in male crickets, Gryllus bimaculatus. AB - When pairs of adult male crickets (Gryllus bimaculatus) that had been housed individually for 7 days were placed together, they fought, and dominant subordinate relationships were formed within 1min. Aggressive behavior by the dominant male was repeated during the period in which the two males were kept together. Immediately after 10min of aggressive interaction, brain serotonin (5 hydroxytryptamine, 5-HT) levels were unchanged in dominant males and significantly reduced in subordinate males. The emission of aggressive song by dominant males is known to be abolished by removal of the wings. All wings were thus removed from male crickets. After 7 days of isolation, pairs of wingless males were placed together. The wingless males fought and formed dominant subordinate relationships within 1min. The wingless, dominant males displayed aggressive behavior. Brain 5-HT levels in the wingless males were reduced immediately after 10min of aggressive interaction, and no significant differences in brain 5-HT levels were detected between the dominant and subordinate males, unlike the case for intact males. These data indicate a difference in brain serotonergic activity between dominant and subordinate male crickets during aggressive interaction, and suggest that aggressive behavior by dominant male crickets rapidly reduce brain 5-HT levels in subordinate ones. Furthermore, the data suggest that aggressive song is responsible for the change in brain 5-HT levels. PMID- 12770184 TI - Characterization of a 212kD protein, released into the host by the larva of the endoparasitoid Chelonus inanitus (Hymenoptera, Braconidae). AB - Previous studies have shown that the larva of Chelonus inanitus plays an essential role in inducing the precocious onset of metamorphosis in polydnavirus/venom-containing Spodoptera littoralis, and that this effect might be due to proteins released by the parasitoid larva into the host. Here we show that a 212kD protein is the predominant protein released in vitro by late first instar parasitoids. This protein does not accumulate anywhere in the parasitoid body. Polyclonal antibodies specific for the 212kD protein were produced and immunoblot analyses revealed that this protein begins to be released by parasitoid larvae when the precocious onset of metamorphosis is induced in the host. Amino acid sequences of the N-terminus and of internal fragments of the 212kD protein were determined and glycoprotein detection showed that this protein is glycosylated. The availability of specific antibodies and the amino acid sequence information should now allow us to clone and express the 212kD protein and to directly study its functional significance. PMID- 12770185 TI - Active regulation of respiration and circulation in pupae of the codling moth (Cydia pomonella). AB - Regulation of autonomic physiological functions has been investigated by means of multisensor electronic methods, including electrocardiographic recording of heartbeat, strain-gauge recording of extracardiac hemocoelic pulsations (EHPs), anemometric recording of air passage through spiracles and respirographic recording of O(2) consumption and CO(2) output. Pupae of Cydia exhibit continuous respiration without remarkable bursts of CO(2). The dorsal vessel of these pupae exhibited regular heartbeat reversals characterized by shorter intervals of faster (forward oriented or anterograde) pulsations and longer intervals of slower (backward oriented or retrograde) peristaltic waves. The periodically repeated EHPs were present during the whole pupal interecdysial period. The internal physiological mechanisms regulating the cardiac (heartbeat) and extracardiac (EHP) pulsations were completely independent for most of the pupal instar. Simultaneous multisensor analysis revealed that the anterograde heartbeat of the dorsal vessel had similar but not identical frequency with EHPs. During advanced pharate adult development, frequency of cardiac and extracardiac pulsation periods profoundly increased until almost uninterrupted pulsation activity towards adult eclosion. At this time, the cardiac and extracardiac pulsations occasionally performed in concert, which enhanced considerably the efficacy of hemolymph circulation in pharate adults with high metabolic rates. The fastest hemolymph flow through the main body cavity was always associated with EHPs and with anterograde heartbeat. Simple physical diffusion of O(2) and CO(2) through spiracles (diffusion theory of insect respiration) does not play a significant role in pupal respiration. Instead, several kinds of regulated, mechanical ventilations of the tracheal system, including EHPs are responsible for effective tracheal ventilation. PMID- 12770186 TI - Enzymological and radiotracer studies of lipid metabolism in the flight-capable and flightless morphs of the wing-polymorphic cricket, Gryllus firmus. AB - The flight-capable morph of the wing-polymorphic cricket, Gryllus firmus, exhibited significantly higher activities of each of five lipogenic enzymes compared with the obligately flightless morph on a standard and a high carbohydrate diet during early adulthood. Similarly, the rate of incorporation of [14C]-acetate into total lipid was higher in the flight-capable morph during this time. By contrast, activities of lipogenic enzymes and rates of lipid biosynthesis, in general, did not differ between morphs on a low nutrient diet during early adulthood. Differences in lipid biosynthesis account for previously documented differences in lipid reserves between morphs on some, but not all, diets. Results of the present and previous studies indicate that increased lipid biosynthesis in the flight capable morph on standard and high-carbohydrate diets constitutes an important adaptation for flight (production of lipid flight fuel). Lipid biosynthesis is negatively correlated with ovarian growth, and may be an important biochemical component of the trade-off between flight capability and ovarian growth in G. firmus. Morphs also differed in activities of three enzymes of lipid catabolism. However, the extent to which variation in activities of these enzymes between morphs results in variation in lipid catabolism is unclear. Finally, the flight-capable morph had a substantially higher activity of alanine aminotransferase in the fat body. Amino acids may be utilized for lipid biosynthesis or energy production to a greater degree in the dispersing morph compared with the oligately flightless morph. This study is the first to document differences in intermediary metabolism that underlie adaptations of morphs of a dispersal-polymorphic species for flight vs. egg production. PMID- 12770187 TI - Photoperiodic time measurement in insects and mites: a critical evaluation of the oscillator-clock hypothesis. AB - The validity of the oscillator-clock hypothesis for photoperiodic time measurement in insects and mites is questioned on the basis of a re interpretation of available experimental evidence. The possible role of the circadian system in photoperiodism in arthropods is critically reviewed. Apart from the outcome of kinetic experiments, based on diel and non-diel light/dark cycles, evidence from various genetic and physiological experiments is discussed in relation to the oscillator-clock hypothesis. The conclusion is that photoperiodic time measurement in insects and mites is performed by a non circadian 'hourglass' clock. Experimental evidence suggests a non-clock role for the circadian system in the photoperiodic mechanism of insects and mites. PMID- 12770188 TI - How drone flies (Eristalis tenax L., Syrphidae, Diptera) use floral guides to locate food sources. AB - In this study we show how inexperienced syrphid flies, Eristalis tenax, orient on artificial flowers by means of floral guides. To test the effect of floral guides such as line and ring markings on the probability and speed of the location of a potential food source, we exploited the spontaneous proboscis reaction triggered by yellow colour stimuli. We tested whether and how fast the flies, when placed on the edge of a circular dummy flower, found a small central yellow spot and touched it with the proboscis extended. The flies found the central yellow spot more often and faster if guide lines from the margin to the yellow spot were present. The effect of guide lines was dependent on the colour of the dummy flower, and independent of the colour of the guide lines, except for yellow guide lines releasing the proboscis reaction. The effect of guide lines was stronger if the yellow spot was hidden in a 2 mm deep depression and thus not as easily visible to the flies. Ring guides had a significant effect on performance only when the intensity of the central yellow spot was low. PMID- 12770189 TI - Juvenile hormone levels in honey bee (Apis mellifera L.) foragers: foraging experience and diurnal variation. AB - A rising blood titer of juvenile hormone (JH) in adult worker honey bees is associated with the shift from working in the hive to foraging. We determined whether the JH increase occurs in anticipation of foraging or whether it is a result of actual foraging experience and/or diurnal changes in exposure to sunlight. We recorded all foraging flights of tagged bees observed at a feeder in a large outdoor flight cage. We measured JH from bees that had taken 1, 3-5, or >100 foraging flights and foragers of indeterminate experience leaving or entering the hive. To study diurnal variation in JH, we sampled foragers every 6h over one day. Titers of JH in foragers were high relative to nurses as in previous studies, suggesting that conditions in the flight cage had no effect on the relationship between foraging behavior and JH. Titers of JH in foragers showed no significant effects of foraging experience, but did show significant diurnal variation. Our results indicate that the high titer of JH in foragers anticipates the onset of foraging and is not affected by foraging experience, but is modulated diurnally. PMID- 12770190 TI - Dietary self-selection and discrimination threshold in wild Anastrepha obliqua females (Diptera, Tephritidae). AB - Experiments were performed on adult wild Anastrepha obliqua females to (I) determine whether there is a critical period during the reproductive phase when the absence of protein in the diet impairs egg production; (II) determine whether the females are able to self-select an optimal diet; and (III) determine the discrimination threshold for protein in newly emerged females as well as in 10 day old females that had either been deprived of protein or not deprived. The results showed that protein ingestion is essential during the preovipositing phase to allow egg production, that females are self-selective and that this behavior changes with age according to the reproductive status, and that protein deprivation strongly alters the discrimination threshold for protein. Newly emerged females have discriminatory thresholds similar to protein-deprived insects. PMID- 12770191 TI - Selective transport of the mulberry leaf urease from the midgut into the larval hemolymph of the silkworm, Bombyx mori. AB - Just before spinning, larvae of the silkworm, Bombyx mori, absorb intact urease of the host plant (mulberry leaf) from the midgut lumen into the hemolymph. In order to investigate whether the transport of the mulberry leaf urease is selective, crude proteins extracted from the mulberry leaves were labeled with biotin and orally administered to the fifth instar larvae. The biotinylated proteins transported into the hemolymph were detected by ligand blotting using streptavidin. When the biotinylated proteins were administered to 5-day-old fifth instar larvae, a strong signal of a biotinylated protein was detected in the hemolymph 2 days after the administration. In contrast, when the biotinylated mulberry leaf proteins were administered to 3-day-old fifth instar larvae, no signal derived from the biotinylated proteins was detected in the hemolymph. The signal weakened when the biotinylated proteins had been immunoprecipitated before administering to the larvae, indicating that the signal came from the mulberry leaf urease. These results show that the transport of the mulberry leaf urease from the midgut into the hemolymph is selective and larval-stage specific. Subsequently, binding assays were carried out to test the binding ability of the mulberry leaf urease to the brush border membrane in the epithelial cells of larval midgut. The urease was not bound to the brush border membrane vesicles (BBMV) from the midgut of 3-day-old fifth instar larvae, while more than 60% of the total amount of incubated urease was bound to the BBMV from the midgut of 6 day-old fifth instar larvae. The urease binding ability of BBMV correlated with the uptake of the mulberry leaf urease. This suggests that a urease binding molecule(s) exists in the BBM of the midgut epithelium, which is involved in the uptake of the mulberry leaf urease. In addition, the uptake of the mulberry leaf urease into the hemolymph was induced by 20-hydroxyecdysone. PMID- 12770192 TI - Differential expression of two HSP70 transcripts in response to cold shock, thermoperiod, and adult diapause in the Colorado potato beetle. AB - Partial clones for two members of Leptinotarsa decemlineata inducible 70kDa heat shock protein family (LdHSP70A and B) were developed using RT-PCR. LdHSP70A, but not LdHSP70B, was upregulated during adult diapause. The ability of L. decemlineata to express these two genes in response to subzero temperatures depended on the thermal history of the beetles. Chilling diapausing beetles increased the rate at which both LdHSP70A and B were expressed following a cold shock at -10 degrees C. Following cold shock at -10 degrees C, LdHSP70B expression peaked after 3h at 15 degrees C for chilled diapausing individuals, decreasing to near background levels by the sixth hour. In contrast, nonchilled diapausing beetles expressed their highest level of LdHSP70B only after 6h at 15 degrees C. Diapausing beetles exposed to a thermoperiod with a mean temperature of either 0 or -2.5 degrees C expressed significantly higher levels of both LdHSP70A and B than beetles exposed to constant 0 or -2.5 degrees C. These results demonstrate that the expression of LdHSP70A and B is differentially regulated in response to diapause and environmental conditioning. PMID- 12770193 TI - The metabolic basis of life history variation: genetic and phenotypic differences in lipid reserves among life history morphs of the wing-polymorphic cricket, Gryllus firmus. AB - The flight-capable morph of the wing-polymorphic cricket, Gryllus firmus, accumulated a substantially greater quantity of total lipid and triglyceride, compared with the obligately flightless morph, during the first five days of adulthood. Increased lipid accumulation in the flight-capable morph was genetically based, and was produced when ovarian growth is substantially reduced in that morph. Temporal changes in lipid levels suggest that the higher triglyceride reserves in the flight-capable morph fed a high-nutrient diet were produced by elevated lipid biosynthesis. By contrast, on a low-nutrient or high carbohydrate diet, increased lipid levels in the flight-capable morph appeared to result primarily from decreased lipid utilization. Increased biosynthesis or retention of triglyceride (the major flight fuel in Gryllus) by the flight capable morph may significantly divert nutrients from egg production and hence may be an important physiological cause of its reduced ovarian growth. The obligately flightless morph allocated a greater proportion of total lipid to phospholipid than did the flight-capable morph. No functionally-significant differences in total lipid or triglyceride were produced between morphs during the last nymphal stadium. A second flightless morph, derived from the flight capable morph by histolysis of flight muscles during adulthood, also had reduced amounts of total lipid and triglyceride but increased ovarian growth compared with the flight capable morph on the standard (high-nutrient) diet. Important qualitative and quantitative aspects of lipid metabolism differ genetically between the flight-capable and flightless morphs of G. firmus and likely contribute importantly to their respective adaptations for flight capability vs. reproduction. This is the first study to document genetically-based differences in energy reserves between morphs of a complex (phase, caste, flight) polymorphism in which morphs also differ genetically in key life history traits. PMID- 12770194 TI - Effects of summer frost exposures on the cold tolerance strategy of a sub Antarctic beetle. AB - The sub-Antarctic beetle Hydromedion sparsutum (Coleoptera, Perimylopidae) is common locally on the island of South Georgia where sub-zero temperatures can be experienced in any month of the year. Larvae were known to be weakly freeze tolerant in summer with a mean supercooling point (SCP) around -4 degrees C and a lower lethal temperature of -10 degrees C (15min exposure). This study investigated the effects of successive freezing exposures on the SCP and subsequent survival of summer acclimatised larvae. The mean SCP of field fresh larvae was -4.2+/-0.2 degrees C with a range from -1.0 to -6.1 degrees C. When larvae were cooled to -6.5 degrees C on 10 occasions at intervals of 30min and one and four days, survival was 44, 70 and 68%, respectively. The 'end of experiment' SCP of larvae surviving 10 exposures at -6.5 degrees C showed distinct changes and patterns from the original field population depending on the interval between exposure. In the 30min interval group, most larvae froze between -6 and -8 degrees C, a depression of up to 6 degrees C from the original sample; all larvae were dead when cooling was continued below the SCP to -12 degrees C. In the one and four day interval groups, most larvae froze above -6 degrees C, showing no change as a result of the 10 exposures at -6.5 degrees C. As with the 30min interval group, some larvae froze below -6 degrees C, but with a wider range, and again, all were dead when cooled to -12 degrees C. However, in the one and four day interval groups, some larvae remained unfrozen when cooled to -12 degrees C, a depression of their individual SCP of at least 6 degrees C, and were alive 24h after cooling. In a further experiment, larvae were cooled to their individual SCP temperature at daily intervals on 10 occasions to ensure that every larva froze every day. Most larvae which showed a depression of their SCP of 2-4 degrees C from their day one value became moribund or died after six or seven freezing events. Survival was highest in larvae with SCPs of -2 to -3 degrees C on day one and which froze at this level on all 10 occasions. The results indicate that in larvae in which the SCP is lowered following sub-zero exposure, the depression of the SCP is greatest in individuals that do not actually freeze. Further, the data suggest that after successive frost exposures in early winter the larval population may become segregated into two sub populations with different overwintering strategies. One group consists of larvae that freeze consistently in the temperature range from -1 to -3 degrees C and can survive multiple freeze-thaw cycles. A second group with lower initial SCPs (around -6 degrees C), or which fall to this level or lower (down to -12 degrees C) after freezing on one or more occasions, are less likely to freeze through extended supercooling, but more likely to die if freezing occurs. PMID- 12770195 TI - Nutritional regulation in nymphs of the German cockroach, Blattella germanica. AB - Synthetic foods varying in protein-carbohydrate ratio and total nutrient concentration were used in food selection experiments to investigate the ingestive and post-ingestive regulation of macronutrients by male and female nymphs of the German cockroach, Blattella germanica (L.) (Dictyoptera: Blattellidae). For both nutrient imbalance and food dilution, food ingestion varied between treatments with the effect that nutrient ingestion was regulated. However, this mechanism was insufficient to compensate for some food dilution treatments. In those cases, the regulation of protein intake was prioritised over that of carbohydrate intake, and two additional regulatory responses were seen. Firstly, cellulose digestion supplemented shortfalls in dietary soluble carbohydrates, and secondly the feeding period within the stadium was prolonged. These ingestive, post-ingestive and developmental responses were orchestrated in such a way that, in all treatments, nutrient gain approached similar levels, despite the variation in food properties. PMID- 12770196 TI - Physiology of diapause and cold hardiness in the overwintering pupae of the fall webworm Hyphantria cunea (Lepidoptera: Arctiidae) in Japan. AB - The fall webworm Hyphantria cunea Drury, which was accidentally introduced to Japan in 1945, overwinters on the ground in pupal diapause. Diapause termination, as indicated by the respiration rate and the period required for adult emergence, began in March and ended in April. Cold hardiness (the ability to survive exposure to -15 degrees C) decreased linearly with diapause development from November to the following April under field conditions. Cold hardiness of diapause pupae (DP) decreased as the acclimation temperature decreased from 15 to -10 degrees C, whereas cold hardiness of non-diapause pupae (NDP) remained high as the acclimation temperature decreased from 5 to -5 degrees C. However, H. cunea in Japan can survive exposure to -5 degrees C for two weeks, whether it is in a diapause or non-diapause state. Trehalose was the main sugar detected in the body, but its level was less than 0.8%. Trehalose levels increased in field collected pupae from January to March. DP accumulated less trehalose than NDP, as the acclimation temperature was decreased from 5 to -5 degrees C. The alanine content in field-collected pupae increased from November to February. Both diapause and low temperature caused an accumulation of alanine. These results suggest that under field conditions, overwintering pupae of H. cunea in Japan do not accumulate high levels of sugars and polyols and do not develop a high level of cold hardiness. Furthermore, DP do not accumulate high levels of sugars and polyols and their ability to survive exposure to -15 degrees C is not greater than that of NDP. The physiological and biochemical bases of diapause in H. cunea from Japan are discussed. PMID- 12770197 TI - Two distinct reproductive strategies are correlated with an ovarian phenotype in co-existing parthenogenetic strains of a parasitic wasp. AB - The question whether different organisms are able to compete for the same resource is of fundamental importance to evolutionary biology. Sympatric co existence of similar species on a single resource has long been claimed to be unstable. However, indirect evidence suggests that parasitic wasps exhibit evolutionarily stable mixtures of life-history strategies. Here we describe genetically distinct strains of a parthenogenetic wasp Venturia canescens, with different ovarian phenotypes that affect egg numbers in oviducts. Wasp females with large egg load search for caterpillars and deposit eggs immediately after host encounter, whereas females with fewer eggs delay parasitism. Since the outcome of interlarval competition within super-parasitized caterpillars depends on the age distribution of competing larvae, the two egg deposition strategies may co-exist under conditions that favor super-parasitism. PMID- 12770198 TI - Drought acclimation confers cold tolerance in the soil collembolan Folsomia candida. AB - It has been noted that both summer drought and sub-zero winter temperatures induce the synthesis of sugars and polyols in invertebrate tissues. This has led several authors to suggest that many of the adaptations, previously viewed as a response to cold, might be part of a more universal desiccation tolerance mechanism. Here we show that acclimation of the soil dwelling collembolan Folsomia candida to a sublethal desiccation stress confers tolerance to cold shock and a significant increase in the molar percent of membrane fatty acids with a mid-chain double bond. These changes in membrane fatty acids are interpreted as conferring a significant reduction in the transition temperature of cell membranes, as would be expected in acclimation to cold, and these changes are therefore interpreted as contributing to the cross-tolerance. Drought acclimation was also shown to trigger the synthesis of the 70kDa family of heat shock proteins (Hsp70). This group of heat shock proteins is implicated in the reestablishment of the normal three-dimensional structure of partially unfolded proteins and therefore are also likely to contribute to the observed cross tolerance. This study provides evidence that the stresses exerted by desiccation and cold at the cellular level have sufficient similarities to induce overlapping adaptations. PMID- 12770199 TI - Distribution and levels of dopamine and its metabolites in brains of reproductive workers in honeybees. AB - To explore the role of dopamine and its metabolites for change of reproductive states of workers in honeybees (Apis mellifera), brain levels of dopamine relative substances were measured and localized in both normal workers and queenless workers. Dopamine and two possible metabolites of dopamine, N acetyldopamine (NADA) and norepinephrine were detected in brain extracts. The brain levels of dopamine, NADA and norepinephrine were positively correlated with ovary development. Individuals with high dopamine levels had high levels of NADA or norepinephrine, suggesting that these metabolites might be involved in the change of reproductive sates of workers. Dopamine was distributed mainly in the protocerebrum, whereas NADA was in both the optic lobes and the protocerebrum. Dopamine levels in each distinct brain regions were higher in queenless workers than in normal workers, whereas there was a higher NADA level in the optic lobes in queenless workers than in normal workers. These results suggest that dopamine might be stored and/or released around the protocerebrum and the deutocerebrum, and also diffuse to the optic lobes where dopamine secretory cells are absent, resulting in high NADA levels in the optic lobes. The different manner of level changes of dopamine and its metabolites in each brain region might cause compound behavioural modulations in reproductive workers. PMID- 12770200 TI - A case for multiple oscillators controlling different circadian rhythms in Drosophila melanogaster. AB - A population of the fruit fly Drosophila melanogaster was raised in periodic light/dark (LD) cycles of 12:12 h for about 35 generations. Eclosion, locomotor activity, and oviposition were found to be rhythmic in these flies, when assayed in constant laboratory conditions where the light intensity, temperature, humidity and other factors which could possibly act as time cue for these flies, were kept constant. These rhythms also entrained to a LD cycle of 12:12 h in the laboratory with each of them adopting a different temporal niche. The free running periods (tau) of the eclosion, locomotor activity and oviposition rhythms were significantly different from each other. The peak of eclosion and the onset of locomotor activity occurred during the light phase of the LD cycle, whereas the peak of oviposition was found to occur during the dark phase of the LD cycle. Based on these results, we conclude that different circadian oscillators control the eclosion, locomotor activity and oviposition rhythms in the fruit fly D. melanogaster. PMID- 12770201 TI - Biphasic effect of low temperature on completion of winter diapause in the onion maggot, Delia antiqua. AB - The effect of low temperature on completion of winter diapause was investigated in the onion maggot, Delia antiqua (Diptera: Anthomyiidae). Diapause was completed under constant diapause-inducing conditions of 15 degrees C and 12L 12D, without any exposure to lower temperature. The pupal period for 50% adult emergence was 117 days. None of the cold treatments at 5.6 degrees C examined in the present study advanced adult emergence; on the contrary, they delayed it. Detailed analyses of the results revealed that diapause development in D. antiqua comprises two phases which differ in sensitivity to low temperature, with the phase shift occurring at around day 60 at 15 degrees C and 12L-12D. In the first phase of diapause development, low temperature (5.6 degrees C) had no effect on diapause development. In the latter phase, by contrast, diapause development was retarded in proportion to the duration of cold treatment. PMID- 12770202 TI - Snowdrop lectin (GNA) has no acute toxic effects on a beneficial insect predator, the 2-spot ladybird (Adalia bipunctata L.). AB - Two-spot ladybird (Adalia bipunctata L.) larvae were fed on aphids (Myzus persicae (Sulz.)) which had been loaded with snowdrop lectin (Galanthus nivalis agglutinin; GNA) by feeding on artificial diet containing the protein. Treatment with GNA significantly decreased the growth of aphids. No acute toxicity of GNA containing aphids towards the ladybird larvae was observed, although there were small effects on development. When fed a fixed number of aphids, larvae exposed to GNA spent longer in the 4th instar, taking 6 extra days to reach pupation; however, retardation of development was not observed in ladybird larvae fed equal weights of aphids. Ladybird larvae fed GNA-containing aphids were found to be 8 15% smaller than controls, but ate a significantly greater number of aphids (approx. 40% to pupation). GNA was shown to be present on the microvilli of the midgut brush border membrane and within gut epithelial cells in ladybird larvae fed on GNA-dosed aphids, although disruption of the brush border was not observed. It is hypothesised that GNA does not have significant direct toxic or adverse effects on developing ladybird larvae, but that the effects observed may be due to the fact that the aphids fed on GNA are compromised and are thus a suboptimal food. PMID- 12770203 TI - The effect of arcelin-1 on the structure of the midgut of bruchid larvae and immunolocalization of the arcelin protein. AB - Some wild accessions of the common bean (Phaseolus vulgaris) contain a family of proteins called arcelins, that are toxic to the larvae of certain bruchid species. Among the six allelic variants of arcelin tested so far, arcelin-5 and arcelin-1 confer the highest level of resistance against the Mexican bean weevil, Zabrotes subfasciatus. The same proteins are not toxic to the bean weevil, Acanthoscelides obtectus, which is also a serious pest of cultivated beans. Arcelins belong to the bean lectin family that includes phytohemaggutinins and alpha-amylase inhibitors. Although homologous to lectins, arcelins are themselves only very weak lectins, and their binding properties have not been clearly established. The toxic properties of arcelins may be related to their recognition of and interaction with the glycoproteins and other constituents of the membranes along the digestive tract of insects. Since arcelin-1 was shown to have growth inhibitory effects for the larvae of Z. subfasciatus but not of A. obtectus, we examined the effect of an arcelin-1 containing diet on the structure of the cells that line the intestinal tract of the larvae of these two bruchid species, and used antibodies against arcelin to examine the distribution of arcelin within the cells and tissues. Here we show that dietary arcelin-1 caused an alteration of the gut structure and the penetration of arcelin into the haemolymph in Z. subfasciatus but not in A. obtectus. These results lead us to suggest that arcelins exert their toxic effect by severely damaging the epithelial cells. PMID- 12770204 TI - Discontinuous gas exchange in the fire ant, Solenopsos invicta Buren: Caste differences and temperature effects. AB - The discontinuous gas exchange cycle (DGC), the cyclic release of CO(2) and uptake of O(2), were investigated in workers and female and male alates of the red imported fire ant, Solenopsis invicta Buren, using real-time CO(2) emission measurement by flow-through respirometry. All S. invicta castes displayed discontinuous emission of CO(2) in the temperature range of 15-25 degrees C, but only male alates and workers exhibited a DGC at 30 degrees C. The closed (C) and flutter (F) periods of the DGC were distinguishable in alates of both sexes at the lowest temperature, but not clearly differentiated in females at temperatures above 15 degrees C, in males above 20 degrees C, or workers at any temperature. DGC frequency increased for all castes as temperature increased, ranging from a low of 0.9+/-0.05 mHz (male alates at 15 degrees C) to 18+/-0.79 mHz (workers at 30 degrees C). O period (or burst) volumes of all castes decreased as temperature increased, and increased with body mass - this mass effect was most pronounced at lower temperatures. Q(10) values for DGC frequency (4.27, 5.81, and 5.62 for workers, female and male alates, respectively) were high compared with Q(10)'s for standard Vdot;(CO(2)). Differences in the salient characteristics of the DGC between castes are presented and discussed, and S. invicta DGC patterns are compared to known values for some other ant species. PMID- 12770205 TI - Induction and development of winter larval diapause in a drosophilid fly, Chymomyza costata. AB - Photoperiodic response during induction of larval hibernal diapause of Chymomyza costata was characterized and the course of diapause development was analyzed in the laboratory. C. costata becomes sensitive to photoperiodic stimuli during an unspecified stage of its early development (embryo, 1st larval instar); the sensitivity gradually increases during the 2nd and early 3rd larval instars and reaches its maximum just before the moment when it abruptly ceases at the age of 15-19 days after oviposition. Diapause intensifies during a period of 2-3 weeks after induction and, later, is maintained without apparent development until death (between 150 and 250 days) under 18 degrees C and a short-day photoperiod (L10:D14, SD). Diapause may be terminated in a horotelic process by exposure to a low temperature (2 degrees C) during which larvae subsequently (1) synchronize their post-diapause development (requires up to 14 days of chilling), (2) lose photoperiodic sensitivity (2 months), and finally (3) terminate diapause (5 months). Alternatively, diapause may be terminated in a tachytelic process by exposure to a high temperature (18 degrees C) and long-day photoperiod (L16:D8, LD) during which no synchronization occurs and pupariation takes place after a mean of 25.2 days (with a broad range from 8 to more than 50 days). Larvae that are transferred from LD to SD during their sensitive period switch their developmental programming from pupariation to diapause. Proliferation of adult primordial structures (imaginal discs, neuroblasts) slows down within 1 day after transfer. In contrast, whole body growth continues for at least 3 days before its rate slows down and matches the rate characteristic for SD conditions. PMID- 12770206 TI - The immune response of the desert locust Schistocerca gregaria during mycosis of the entomopathogenic fungus, Metarhizium anisopliae var acridum. AB - Topical application of Metarhizium anisopliae var acridum to the desert locust Schistocerca gregaria resulted in changes in the biochemistry and antimicrobial defenses of the haemolymph. M. anisopliae var acridum colonized the host haemolymph from day two post application. The haemocytes did not attach to, phagocytose or nodulate elements of the fungus. However, the presence of the fungus appeared to stimulate hemocyte aggregation over the first few days of mycosis though the number of aggregates declined subsequently. The total hemocyte count increased two days after application, indicating an overall stimulation of the immune system, but declined to a value below that for uninoculated controls by day four. The differential haemocyte count showed that the initial increase in total haemocyte count was primarily due to a larger number of coagulocytes. After day two consistent declines in cell number were observed for all haemocyte classes in mycosed insects. The activity of the enzyme, phenoloxidase, decreased during the course of infection. However, the converse was true for prophenoloxidase. Lysozyme levels were significantly smaller in infected than control locusts. There was a significant correlation between lysozyme and PO activities when data from mycosed and control insects were combined. The total protein content of the haemolymph decreased during the course of infection. PMID- 12770207 TI - Anaesthetising Drosophila for behavioural studies. AB - The detrimental effect of anaesthesia by chilling or CO(2) on the mating behaviour of Drosophila melanogaster was investigated. Both agents significantly increased copulation latency, even when flies were given 20 h to recover from anaesthesia. CO(2) anaesthesia increased copulation latency more than anaesthesia by chilling. Delivery of a mechanical shock to the flies immediately before testing also increased copulation latency. These experiments demonstrate the sensitivity of insect behaviour to disruption by anaesthesia or rough handling. It is preferable to avoid anaesthetising flies that are to be used in behavioural studies. PMID- 12770208 TI - Chemistry, ontogeny, and role of pygidial gland secretions of the vinegaroon Mastigoproctus giganteus (Arachnida: Uropygi). AB - Pygidial gland secretions of the vinegaroon, Mastigoproctus giganteus, consist of 26% water and 74% short-chained fatty acids, with acetic acid and octanoic acids the dominant acids. The organic composition of the secretion consists of 83% acetic acid, 15% octanoic acid, 1% (Z)-5-octenoic acid, 0.0002% of the corresponding (E)-isomer, 0.11% heptanoic acid, 0.02% hexanoic acid, 0.007% decanoic acid, and in some individuals a trace of 2-nonanone. Formic acid is absent from the secretion. Glandular composition, especially in the concentrations of the minor components, varies dramatically among individuals and shows no age pattern differences among the four free-living subadult instars, or sex differences between adult males and females. Each individual exhibits its own specific blend of components which is identical in the two separate and different sized glands. The secretions act solely as allomones against potential predators, and do not act as pheromones, or elicit a communicative or behavioral response. Behavioral tests using live animals or a synthetic blend of vinegaroon secretion revealed that the secretion is maximally effective on sensitive respiratory, visual, or sensory systems of potential predators and is ineffective, or nearly so, when contact is restricted to inside the predator's mouth. Thus, the effectiveness of the secretion apparently results from its delivery as a spray of fine droplets, rather than as an exudate or vapor. PMID- 12770209 TI - Stress-reactivity of a Drosophila melanogaster strain with impaired juvenile hormone action. AB - Met(27) is a null allele of the Methoprene-tolerant gene of D. melanogaster that shows resistance to the toxic effects of both juvenile hormone (JH) and a JH analog, methoprene. The mechanism of resistance appears to be altered JH reception. We measured fertility, JH-hydrolyzing activity, and dopamine (DA) levels in Met(27) and Met(+) flies under normal (25 degrees C) and heat-stress (38 degrees C) conditions. We show that under normal conditions Met(27) females have JH-hydrolyzing activity and fertility lower than Met(+), but DA content did not differ between the two strains. At 38 degrees C Met(27) flies show no impairment in JH-hydrolyzing activity in response to stress, but they do show lower DA levels and impaired reproduction. The results with Met(27) are consistent with the previous hypothesis that the alteration in fertility that follows heat stress in D. melanogaster could result from alteration in the JH endocrine system. PMID- 12770210 TI - Ecdysteroid levels/profiles of the parasitoid wasp, Diapetimorpha introita, reared on its host, Spodoptera frugiperda and on an artificial diet. AB - Diapetimorpha introita is an ichneumonid ectoparasitoid of the fall armyworm, Spodoptera frugiperda. Since it has been reported that D. introita wasps reared on an artificial diet exhibit a significantly lower percentage of adult eclosion and fecundity than host-reared wasps, this study was undertaken to elucidate the factors responsible for the reduced viability observed in diet-reared wasps. A system of markers has been devised to track the development (from the initiation of cocooning through adult eclosion) of D. introita. Although wasps reared on artificial diet developed more slowly than did those reared on host pupae, both diet- and host-reared wasps passed through the same stages of development - the eyes enlarged and moved backward, the gut was purged and upon ecdysis the exarate pupa emerged. The thorax was the first to darken, followed by the head and then the abdomen. Pharate pupal formation occurred before gut purge. Two peaks of hemolymph ecdysteroids were observed, one in wasps in which gut purge was almost complete and the second in day-2 exarate pupae. Ecdysone and 20-hydroxyecdysone were the major ecdysteroids present in hemolymph sampled at these times. Small quantities of 20,26-dihydroxyecdysone, polar ecdysteroids and/or possibly 26 hydroxyecdysone were also present. In six stages of development, hemolymph ecdysteroid titers were significantly higher in host-reared than in diet-reared wasps (Eye 1, Eye 2, Gut Purge 2, Pharate Pupa, Head/Thorax Dark, and Abdomen Dark). Relatively high percentages of mortality were observed in diet-reared wasps in four of these stages and in two others which occurred in close proximity to one of the stages, the Abdomen Dark stage. Thus, insufficient ecdysteroid in the hemolymph may be responsible, in part, for the relatively high percentage of mortality that occurred in wasps reared on an artificial diet. PMID- 12770211 TI - Growth-promoting effects of ecdysteroids and juvenile hormone on in vitro development of the larval endoparasitoid, Venturia canescens (Hymenoptera: Ichneumonidae). AB - We previously reported that lipophorin, fetal bovine serum (FBS), and 20 hydroxyecdysone (20-HE) are essential for the development of the larval endoparasitoid Venturia canescens larvae in vitro. The present study was undertaken to determine the optimal concentrations of those three substances in the MGM-450 medium, and to examine the hormonal effects of ecdysteroids and juvenile hormone (JH) on the development of the parasitoid larvae in vitro. When the culture was started with embryos at the post-germband stage, concentrations of 3 mg/ml of lipophorin and 20% of FBS were most suitable for the development of the parasitoid. The growth-promoting effect of 20-HE increased in a concentration dependent manner and peaked at a concentration of 1 &mgr;g/ml. Excess concentration led to malformations of the larvae. Three other ecdysteroids, ecdysone, 2-deoxy-20-hydroxyecdysone, and polypodine B had the same effect, although their activity was lower than that of 20-HE. Cholesterol had no effect; most larvae failed to develop. When the medium was supplemented with JH, the duration of the developmental period was significantly shortened, but this hormone was not found to be essential. PMID- 12770212 TI - Absence of insect juvenile hormones in the American dog tick, Dermacentor variabilis (Say) (Acari:Ixodidae), and in Ornithodoros parkeri Cooley (Acari:Argasidae). AB - Synganglia, salivary gland, midgut, ovary, fat body and muscle alone and in combination from the ixodid tick, Dermacentor variabilis (Say), or the argasid tick, Ornithodoros parkeri Cooley, were incubated in vitro in separate experiments with L-[methyl-(3)H]methionine and farnesoic acid or with [1 (14)C]acetate. Life stages examined in D. variabilis were 3 and 72 h old (after ecdysis) unfed nymphs, partially fed nymphs (18 and 72 h after attachment to the host), fully engorged nymphs (2 d after detachment from host), 3 and 72 h old (after eclosion) unfed females, partially fed unmated females (12-168 h after attachment to host) and mated replete females (2 d after detachment from the host). Those from O. parkeri were third and fourth stadium nymphs and female O. parkeri, 1-2 d after detachment. Corpora allata from Diploptera punctata, Periplaneta americana and Gromphadorina portentosa were used as positive controls in these experiments. No farnesol, methyl farnesoate, JH I, JH II, JH III, or JHIII bisepoxide was detected by radio HPLC from any tick analysis while JH III, methyl farnesoate, and farnesol were detected in the positive controls. To examine further for the presence of a tick, insect-juvenilizing agent, Galleria pupal-cuticle bioassays were conducted on lipid extracts from 10 and 15 d old eggs, unfed larvae (1-5 d after ecdysis), unfed nymphs (1-7 d after ecdysis), and partially fed, unmated female adults (completed slow feeding phase) of D. variabilis. Whole body extracts of fourth stadium D. punctata and JH III standard were used as positive controls. No juvenilizing activity in any of the tick extracts could be detected. Electron impact, gas chromatography-mass spectrometry of hemolymph extracts from fed, virgin (forcibly detached 7 d after attachment) and mated, replete (allowed to drop naturally) D. variabilis and fully engorged (1-2 d after detachment) O. parkeri females also failed to identify the common insect juvenile hormones. The same procedures were successful in the identification of JH III in hemolymph of fourth stadium D. punctata. Last stadium nymphal (female) O. parkeri implanted with synganglia from second nymphal instars underwent normal eclosion to the adult. The above studies in toto suggest that D. variabilis and O. parkeri do not have the ability to make the common insect juvenile hormones, and these juvenile hormones do not regulate tick metamorphosis or reproduction as hypothesized in the literature. PMID- 12770213 TI - Artificial insemination using cryopreserved sperm in the silkworm, Bombyx mori. AB - We report in this paper that female moths artificially inseminated with cryopreserved sperm (-196 degrees C) could oviposit eggs when the sperm was preserved for 356days, and that the fertilization rate and the number of eggs laid were almost equivalent to those obtained in normally mated moths. The optimal cooling rate for sperm freezing was 5-65 degrees C/min for maintaining a high fertility of sperm. The simple and reliable method of cryopreservation was to put the semen first in a deep freezer at -80 degrees C and thereafter put them in liquid nitrogen. When female moths of 'white 2' egg-color mutant strain were inseminated with a mixture of frozen-thawed sperm from males of normal-colored egg strain and non-frozen sperm from males of the 'white 2', female moths deposited a majority of 'white 2' eggs and a very small number of eggs of normal color. The result shows that there was a competitive fertilization of sperm between the two strains of the silkworm, and that sperm fertility was reduced to a considerable extent by freezing at -196 degrees C. These results may contribute not only to basic studies on fertilization in Lepidoptera but also to the development of long-term preservation procedure of genetic resources by using cryopreserved sperm of Bombyx mori. PMID- 12770214 TI - Sodium channels in central neurons of the tobacco budworm, Heliothis virescens: basic properties and modification by scorpion toxins. AB - Voltage-activated sodium channels in central neurons of larval and adult Heliothis virescens were characterized using whole-cell patch clamp techniques. Macroscopic currents showing rapid activation and inactivation kinetics were uniformly sensitive to tetrodotoxin (IC(50)=1.9nM). Currents began to activate at voltage steps to -45mV and reached half maximal at -30mV. Fast inactivation was evident at voltages as negative as -75mV and reached half maximal at -50mV. Full recovery from inactivation occurred within 1 to 2ms. Currents in larval neurons exhibited similar properties to those of adult neurons, except for the rate of fast inactivation (t(1)), which was significantly slower in larval neurons. The biophysical properties of sodium channels remained unchanged for up to 3days in culture. Two insecticidal neurotoxins, LqhalphaIT and AaIT, produced distinctly different modifications of H. virescens sodium channels. LqhalphaIT slowed channel inactivation, while AaIT specifically shifted voltage-dependent activation to more negative potentials. Overall, the results indicate that sodium channels in H. virescens neurons exhibit biophysical characteristics similar to those of vertebrates, yet possess pharmacological uniqueness with respect to scorpion toxin modification. PMID- 12770215 TI - Larval diapause duration and fat metabolism in three geographical strains of the blow fly, Calliphora vicina. AB - Diapausing larvae of the blow fly, Calliphora vicina, from three geographical strains exposed, as adults, to short days, were maintained under identical conditions (darkness, 11-12 degrees C) and examined for changes in wet weight, dry weight, water and fat content during diapause development to the emergence of post-diapause adults. Larvae produced by flies originating from northern Finland (Nallikari, 65 degrees N) showed a longer, more intense, diapause than those from localities further south (Edinburgh, Scotland, 55 degrees N and Barga, Italy, 44 degrees N), but all three strains showed similar rates of loss of the parameters measured. This was also the case for post-diapause adults, flies of the Barga strain with its relatively short diapause emerging with greater residual fat reserves than flies from the Edinburgh or Nallikari strains with their more protracted diapause. It was concluded that the rates of water and fat loss were functions of the conditions used for diapause larval maintenance (probably temperature) rather than the maternally programmed degree of diapause incidence, or of its 'depth' or 'intensity'. PMID- 12770216 TI - Effects of tissue extracts on oviduct contraction in the migratory grasshopper, Melanoplus sanguinipes. AB - The isolated oviduct of the migratory grasshopper, Melanoplus sanguinipes, shows a myogenic, rhythmic pattern of contraction. The pattern of contraction can be modified by treatment with hemolymph or extracts from a variety of tissues from animals of differing age, sex, and mated status. Extracts of oviducts and ventral nerve cord (VNC), as well as hemolymph, from both virgin and mated females were almost always stimulating, though their effect on frequency, amplitude and/or tonus varied. In contrast, whereas extracts of spermatheca and brain from mated females were stimulating, extracts of these tissues from virgin females inhibited contraction. All male material tested (long hyaline tubules (LHT), the male accessory gland complex less the LHT, testes, brain, VNC and hemolymph) stimulated oviduct contraction in a dose-dependent manner, usually enhancing at least two of frequency, amplitude and tonus. However, oviposition-stimulating protein, a purified product of the LHT, provoked only an increase in frequency when applied to the oviduct. LHT extract modulated the effects of virgin female tissue extracts, always in a stimulatory manner. PMID- 12770217 TI - Brain control of mating behavior in the male cricket Gryllus bimaculatus DeGeer: the center for inhibition of copulation actions. AB - We re-examined the functional role of the brain and suboesophageal ganglion (SOG) in inhibiting mating behavior in the male cricket Gryllus bimaculatus DeGeer. Experiments were conducted by using mimetic stimulation to elicit copulation actions. To induce a change in the male internal state from a sexually responsive state to a sexually unresponsive state in the mating stage, noxious stimulation, head injury and leg pinching were used. Males that sustained a head injury became sexually unresponsive but became responsive as soon as the brain was removed, while males that underwent the same treatment remained unresponsive after the SOG was destroyed by sectioning. The inhibitory effects of the brain were also demonstrated by the fact that further removal of the SOG in the decerebrated males did not change their copulatory responsiveness, while removal of the brain in SOG-sectioned males markedly increased copulatory responsiveness. Furthermore, decerebrated males did not become sexually unresponsive by leg pinching, while SOG-sectioned males that had recovered sexual responsivenss, did. These results suggest that the brain, and not the SOG, plays a key role in the inhibition of copulation in the male cricket during the mating stage. PMID- 12770218 TI - Brain control of mating behavior in the male cricket Gryllus bimaculatus DeGeer: brain neurons responsible for inhibition of copulation actions. AB - We localized brain neurons responsible for the inhibition of copulation actions in the male cricket, Gryllus bimaculatus. Males with one connective-cut between the brain and the terminal abdominal ganglion (TAG) were able to perform normal reproductive behaviors such as courtship, copulation and spermatophore protrusion. However, copulation response tests using a female dummy showed that they failed to exhibit a transient inhibitory state after application of noxious stimulation in the mating stage and to exhibit a tonic inhibitory state in the post-copulatory sexual refractory stage. A partial cut of a single connective (hemilateral connective-cut) combined with the copulation response test and an axonal backfilling demonstrated that approximately 40 brain neurons (per one connective), whose somata were located in the posterior region of the protocerebrum and mainly contralaterally to their descending axons running through the dorso-medial part of the connective, were candidates for the inhibition of copulation actions, at least in the mating stage. In addition, we report here for the first time, that copulation actions can be elicited shortly after copulation even in intact males. This unusual behavior was frequently observed in fledglings after the final molt. The results are discussed from the perspective of the operation of the brain inhibition system on the central pattern generator (CPG) for copulation. PMID- 12770219 TI - A comparative analysis of metabolic rate in six Scarabaeus species (Coleoptera: Scarabaeidae) from southern Africa: further caveats when inferring adaptation. AB - Alterations in VO(2) or VCO(2) are amongst the more polemical physiological adaptations ascribed to insects. Generally, metabolic rate is thought to be lowered in response to arid conditions, and elevated in species from cold environments compared to their more temperate relatives. However, most studies have rarely addressed the influence of both environmental factors in unison. To this end, standard metabolic rate and its temperature dependence were measured (at 4 degrees C intervals from 16 to 32 degrees C) in six Scarabaeus dung beetle species (three flightless, three volant) from a variety of habitats (warm, arid to cool, mesic) in southern Africa using flow-through respirometry. Mass specific VCO(2) varied from 0.0158 ml g(-1) h(-1) at 16 degrees C to 0.1839 ml g (-1) h( 1) at 32 degrees C. The slopes of the rate temperature curves were similar for all species (Q(10)s of 2.14-2.84), although the intercepts differed significantly in the direction (warm arid to cool mesic): S. gariepinus>AMP>/=ATP. However, the salt receptor cell could respond significantly not only to GDP but various nucleoside 5'-diphosphates, nucleoside 5'-monophosphates, cyclic nucleotides and thiamine diphosphate. These results clearly suggest that the specificity of the receptor site reacting with nucleotide in the sugar receptor cell is very different from that in the salt receptor cell. PMID- 12770373 TI - Disturbance of adult eclosion by fenoxycarb in the silkworm, Bombyx mori. AB - Fenoxycarb treatment before and after pupal ecdysis of Bombyx mori disturbed adult eclosion and the animals were unable to escape from the pupal exuviae. This effect of fenoxycarb was dose and time dependent with the highest sensitivity around the pupal ecdysis. The sensitivity rapidly diminished within 20 hours of pupal ecdysis. Twenty-hydroxyecdysone (20E) produced similar effects. Fenoxycarb injection at the pupal ecdysis induced higher ecdysteroid production by the prothoracic glands and higher PTTH-secretory activity in the brain-corpora cardiaca-corpora allata complexes. As a result, the fenoxycarb treated pupae contained higher ecdysteroid titres in the haemolymph. Both the fenoxycarb and the 20E treatments resulted in the lack of development of the rectum in pharate adults. This was the main cause of high ecdysteroid titres in the pharate adult stage. This effect was mimicked by either removal of the rectum early in the pharate adult stage or a surgical extirpation of the hindgut at the time of pupal ecdysis. These results suggest that the disturbance of adult eclosion by fenoxycarb is due in part to the inability of the formation of the rectum in the pharate adult stage. PMID- 12770374 TI - Use of Rb(+) and Br(-) as tracers for investigating ion transport by X-ray microanalysis in the Malpighian tubules of the black field cricket Teleogryllus oceanicus. AB - Substitution of Rb(+) for K(+) in the incubation saline for in vitro preparations of Malpighian tubules had little effect on tubule function. Secretion rates increased by 10% for whole tubules, 9% for distal segments and 10% for main segments. In the secreted fluids Rb(+) almost completely replaced K(+). Within the cells of the main segment of the tubules Rb replaced the majority of the intracellular K. Treatment by ouabain in Rb saline resulted in a considerable increase in intracellular Na and Cl concentrations but no change in Rb concentration. This suggests that Rb(+) did not enter the cell via Na K ATPase and that the latter was not directly involved in Rb(+) secretion and by inference K(+) secretion. Substitution of Br(-) for Cl(-) in the incubation saline resulted in a 30% reduction in secretion rate from the distal segments but only a 10% reduction for the main segment. Cl(-) was almost completely replaced by Br(-) in fluid from both main and distal segments. In cells of the main segment the intracellular concentration of Br(-) did not exceed 30mmol kg(-1) dry weight and the Cl(-) concentration was unchanged in the apical region of the cell and increased in the basal region. These data suggest that Br(-) was transported across the tubule epithelium by a paracellular route and that the basal cell membrane is relatively impermeable to Cl(-). By inference Cl(-) may also be transported by a paracellular route. PMID- 12770375 TI - Biochemical aspects of flight and flightlessness in Gryllus: flight fuels, enzyme activities and electrophoretic profiles of flight muscles from flight-capable and flightless morphs. AB - Female Gryllus assimilis subjected to 4.5-7.7h continuous tethered flight had significantly lower amounts of total lipid, triglyceride and total soluble carbohydrate compared with unflown controls. A much greater amount of total lipid (6.3mg) was used during flight compared with carbohydrate (0.14mg). Flown individuals also had substantially reduced amounts of injected, radiolabeled [(14)C]-oleic acid. Activities of lipid, carbohydrate and amino acid catabolizing enzymes in flight muscles of G. assimilis and its wing-polymorphic congener, G. firmus, were very similar to activities in insects which primarily utilize lipid to power flight. By contrast, enzyme activities were very different from those in insects which primarily or exclusively use carbohydrate or proline as a flight fuel. These results strongly implicate lipid as the major flight fuel in Gryllus. Previous studies have shown that lipid levels are higher in flight-capable (long winged) G. firmus that have small ovaries compared with flightless (short-winged) females that have large ovaries. Results of the present and previous studies collectively indicate that elevated lipid in long-winged G. firmus represents an energetic cost of flight capability which reduces (trade-offs with) reproduction in Gryllus. In G. firmus, mass-specific activities of nearly all enzymes were considerably reduced in underdeveloped, and to a lesser degree in histolyzed muscle, compared with fully-developed flight muscle. An important exception was alanine aminotransferase, whose activity was the highest in histolyzed muscle, and which may be involved in the catabolism of amino acids derived from muscle degradation. Despite the dramatic differences in enzyme activity, electrophoretic profiles of soluble flight-muscle proteins differed only subtly between fully developed and underdeveloped or histolyzed flight muscles. PMID- 12770376 TI - Atypical vitellins in ponerine ants (Formicidae: Hymenoptera). AB - Higher hymenopteran vitellogenin/vitellins have been characterized as containing one large apoprotein. We show that in the ant subfamily Ponerinae, species in the tribes Odontomachini, Platythyrini, and Amblyoponini, also have a vitellin with this simple structure, containing a single apoprotein of 180-190kDa. Species in tribes Ponerini and Ectatommini, however, have vitellins containing multiple subunits. The size and number of the subunits varies, with differences even among species within the same genus. This is the first report of diversity in vitellogenin structure in the higher Hymenoptera. Vitellin and vitellogenin in Harpegnathos saltator (Ponerini) contain two large subunits of about 165kDa and two small subunits of about 45 and 43kDa. N-terminal sequence analysis suggests that provitellogenin is cleaved at two different sites, producing two large and two small subunits differing slightly in size. Diversity of vitellin types in Ponerini and Ectatommini is similar to that found in the more primitive tenthredinoid sawflies (Hymenoptera, Symphyta), and may indicate polyphyly in the Ponerinae. Insect vitellogenins and yolk proteins show considerably more diversity than originally believed, and the possibility of the functional significance of these differences should be considered. PMID- 12770377 TI - Effects of photoperiod and temperature on seasonal morph development and diapause egg oviposition in a bivoltine race (Daizo) of the silkmoth, Bombyx mori L. AB - A bivoltine race (Daizo) of the silkmoth, Bombyx mori, has two types of adults which are classified on the basis of the number of dark-brown and brown scales distributed along the outer lines of the anterior wings. Development of these two types of adults is determined by photoperiod and temperature experienced during the embryonic and larval stages. The two types of adults are thought to be seasonal (summer and autumn) morphs. All adults developed under short days at low temperature (15 degrees C and 23 degrees ) were classified as summer morphs and oviposited only nondiapause eggs (ND-eggs), and those that developed under long days at high temperature (28 degrees C and 25 degrees C) were autumn morphs ovipositing only diapause eggs (D-eggs), only ND-eggs or a mixture of D- and ND eggs. Larval exposure to long days at 25 degrees C shifted the wing pattern towards the autumn morphs and decreased the incidence of female adults ovipositing only D-eggs. Larval exposure to long days elicits an opposite effect on D-egg oviposition to that observed when long days are received only during the embryonic stage. PMID- 12770378 TI - Sperm reflux and its role in multiple mating in males of a butterfly Polygonia c aureum Linnaeus (Lepidoptera: Nymphalidae). AB - The relationship between sperm quantity in the duplex and that in the vasa deferentia was examined in the Asian comma butterfly, Polygonia c-aureum. In virgin males, the number of eupyrene sperm bundles in the duplex increased linearly with age, whereas that in the vasa deferentia was consistently small. However, numerous sperm were found in the vasa deferentia of males immediately after mating. The number of eupyrene sperm bundles in the vasa deferentia after mating significantly increased with age and with increasing the time interval between matings. From these and other results, it was suggested that some sperm in the duplex were moved back to the vasa deferentia during mating, and that such sperm reflux provides a means to save sperm for multiple mating. PMID- 12770379 TI - Monoclonal antibodies bind distinct classes of hemocytes in the moth Pseudoplusia includens. AB - Insect hemocytes have historically been identified on the basis of morphology, ultrastructure and hypothesized function. Among insects in the order Lepidoptera, five hemocyte classes are usually recognized: granular cells, plasmatocytes, spherule cells, oenocytoids and prohemocytes. We have generated a panel of monoclonal antibodies (mAbs) against hemocytes of the moth Pseudoplusia includens. In this study, hemocyte identification using 16 different mAbs was compared to identification methods using morphological characters. Three main categories of mAb binding activity were identified: (1) mAbs that specifically labeled only one morphological class of hemocytes, (2) mAbs that labeled granular cells and spherule cells, and (3) mAbs that labeled plasmatocytes and oenocytoids. With one exception, none of the antibodies bound to other tissues in P. includens. However, certain mAbs that specifically labeled granular cells and/or spherule cells in separated hemocyte populations also labeled plasmatocytes co-cultured with granular cells or cultured in granular cell conditioned medium. Overall, our results suggest that granular cells are antigenically related to spherule cells, and that plasmatocytes are antigenically related to oenocytoids. The use of mAbs as hemocyte markers are discussed. PMID- 12770380 TI - Drosophila melanogaster sex peptide stimulates juvenile hormone synthesis and depresses sex pheromone production in Helicoverpa armigera. AB - Previous studies demonstrate that virgin female adult Helicoverpa armigera (Lepidoptera: Noctuidae) moths exhibit calling behaviour and produce sex pheromone in scotophase from the day after emergence, and that mating turns off both of these pre-mating activities. In the fruit fly Drosophila melanogaster, a product of the male accessory glands, termed sex peptide (SP), has been identified as being responsible for suppressing female receptivity after transfer to the female genital tract during mating. Juvenile hormone (JH) production is activated in the D. melanogaster corpus allatum (CA) by SP in vitro. We herein demonstrate cross-reactivity of D. melanogaster SP in the H. armigera moth: JH production in photophase virgin female moth CA in vitro is directly activated in a dose-dependent manner by synthetic D. melanogaster SP, and concurrently inhibits pheromone biosynthesis activating neuropeptide (PBAN)-activated pheromone production by isolated pheromone glands of virgin females. Control peptides (locust adipokinetic hormone, AKH-I, and human corticotropin, ACTH) do not inhibit in vitro pheromone biosynthesis. Moreover, SP injected into virgin H. armigera females, decapitated 24 h after eclosion, or into scotophase virgin females, suppresses pheromone production. In the light of these results, we hypothesize the presumptive existence of a SP-like factor among the peptides transmitted to female H. armigera during copulation, inducing an increased level of JH production and depressing the levels of pheromone produced thereafter. PMID- 12770381 TI - Expression of the juvenile hormone esterase gene in the Colorado potato beetle, Leptinotarsa decemlineata: photoperiodic and juvenile hormone analog response. AB - Metamorphosis and reproduction in insects are controlled by juvenile hormone (JH). One of the factors, which regulate the JH titer in the hemolymph, is the activity of juvenile hormone esterase (JHE). JHE from the Colorado potato beetle, Leptinotarsa decemlineata, consists of two 57kDa subunits. In this study, the JHE cDNA was used as a probe to examine where and when the gene is transcribed as well as how gene expression responds to photoperiodic treatment and to topical application with a JH analog, pyriproxyfen. JHE transcripts were almost exclusively found in RNA extracts from fat body tissue in both larvae and adults. JHE-mRNA levels in the fat body correlated positively with levels of JHE activity in the hemolymph. In the last larval instar, high levels of JHE-mRNA were found in the feeding stage. In adults, reared under short-day conditions, JHE-mRNA levels were high between day 2 and day 9, which correlated with high JHE activity in the hemolymph. During these conditions, the JH titer decreases in preparation for pupation and diapause, respectively. The JHE-mRNA levels and JHE activity in the hemolymph were higher in short-day than in reproductive long-day adults. If the JH analog pyriproxyfen was applied to animals of the last larval instar on day 0 or day 3, JHE gene expression was enhanced. In contrast, if pyriproxyfen was applied to short-day adults on day 1 or day 4, the mRNA levels and the JHE activity in the hemolymph were suppressed to levels similar to those found in long-day adults. Thus, transcription of JHE is dependent on developmental stage, tissue, photoperiod and the level of its substrate JH. PMID- 12770382 TI - Purification and characterization of an oviposition-stimulating protein of the long hyaline tubules in the male migratory grasshopper, Melanoplus sanguinipes. AB - An oviposition-stimulating protein (OSP) was isolated and purified from the long hyaline tubules of the male accessory gland complex in the migratory grasshopper, Melanoplus sanguinipes. Gel filtration of the native OSP, using Sephadex G-100, indicates its molecular weight to be about 60000Da with the oviposition stimulating activity while sodium dodecyl sulphate-polyacrylamide gel electrophoresis shows that the OSP comprises two subunits, each with a molecular weight of 30000Da. The purified OSP appears as a single symmetric peak on fast performance liquid chromatography using Mono Q. Isoelectric focusing of the OSP indicates an apparent pI of 5.5. Injection of the OSP induces oviposition in about 70% of ovulated virgin females within 48h. Stimulation of oviposition can be blocked by a polyclonal antibody raised against the OSP 30000Da subunits. Amino acid analysis of the dimer and its subunits shows a comparatively high content of aspartic acid/asparagine (14.8%) as well as leucine (12.2%) and glutamic acid/glutamine (12.0%). The N-terminal 21 amino acid sequence of the OSP shows little similarity to known peptides. Immunoreactivity with the anti-OSP antibody was observed in the viscous secretion, spermatheca, and the egg-pod froth of mated females, confirming transfer of the OSP from male to female during copulation. PMID- 12770383 TI - Effects of pH on feeding responses in the apple maggot fly, Rhagoletis pomonella (Diptera: Tephritidae). AB - Phagostimulatory effects of pH values of sucrose on Rhagoletis pomonella adults were studied in the laboratory. Flies were standardized for age, diet and food deprivation. Two presentation schemes were employed. The first varied pH value (3.0-10.0) with sucrose concentration kept constant at 40%. The second varied both sucrose concentration (8%, 24% and 40%) and pH value (5.0-8.0). Fly feeding propensity was evaluated by observation of fly acceptance or rejection of sucrose and duration of feeding. When tested on red wooden spheres treated with 40% sucrose, fly feeding acceptance was significantly greater when pH ranged from 5.0 to 8.0, and duration of feeding was significantly longer at pH 6.0-7.0. At pH /=8.0, feeding propensity was significantly reduced. Decrease in sucrose concentration significantly increased fly sensitivity to pH. Males were more responsive to varying pH than females. The sucrose pH shown to stimulate maximal feeding response was 6.4. Such information is relevant to formulation improvement of a coating mixture of sucrose and insecticide applied to red spheres as part of apple maggot fly control programs. PMID- 12770384 TI - A baculovirus enhancin alters the permeability of a mucosal midgut peritrophic matrix from lepidopteran larvae. AB - The peritrophic matrix (PM) in lepidopterous larvae may function as a defensive barrier against ingested viral pathogens. PMs isolated from Trichoplusia ni and Pseudaletia unipuncta larvae, were treated with a baculovirus-encoded metalloprotease (enhancin) from Trichoplusia ni granulosis virus (TnGV) and their in vitro permeability to blue dextran and fluorescent-labelled Autographa californica nuclear polyhedrosis virus (AcMNPV) was determined using a dual chamber permeability apparatus. Incubation of T. ni PMs with 0.0, 0.5, 1.0, and 2.0mg/ml enhancin resulted in a blue dextran 2000 flux of 4.4, 6.3, 9.9, and 15.6&mgr;g/mm(2)/h, respectively. In addition, T. ni PMs treated with enhancin were found to be significantly more permeable to fluorescent-labelled AcMNPV than non-treated control PMs. The permeability of T. ni PMs treated with 3.0mg/ml enhancin was 0.017 cumulative percent crossing/mm(2)/h, whereas the permeability of the control PM was below the detectable limit. Similarly, enhancin treatment greatly increased the permeability of P. unipuncta PMs to AcMNPV. These results provide evidence that the PM from two lepidopteran species can block the passage of baculovirions across this matrix thus reducing the probability of larval infection. Furthermore, these results support the hypothesis that enhancin facilitates NPV infection of larvae by altering the permeability of the PM. PMID- 12770385 TI - Seasonal change of the thermal response in relation to myo-inositol metabolism in adults of Aulacophora nigripennis (Coleoptera Chrysomelidae). AB - The thermal response of Aulacophora nigripennis adults in relation to myo inositol metabolism changed seasonally. Myo-inositol accumulation was stimulated at 15 and 5 degrees C in October (autumn), but only 5 degrees C was effective for the accumulation in December (early winter). In February (mid winter), myo inositol degradation was prominent at both temperatures. Myo-inositol metabolism of this beetle thus gradually shifts from synthesis to degradation as the season progresses. This may be related to the progress of diapause development and may contribute to the seasonal profile of the myo-inositol pool in a natural population. Post-diapause adults in March (spring) and June (early summer) regained the ability for myo-inositol accumulation at 5 degrees C. Thus, there are at least two types of myo-inositol accumulation in this beetle; one is under the control of diapause and the other responds directly to environmental conditions independently of diapause. PMID- 12770386 TI - Development of the parasitoid, Cotesia rubecula (Hymenoptera: Braconidae) in Pieris rapae and Pieris brassicae (Lepidoptera: Pieridae): evidence for host regulation. AB - Several recent models examining the developmental strategies of parasitoids attacking hosts which continue feeding and growing after parasitism (=koinobiont parasitoids) assume that host quality is a non-linear function of host size at oviposition. We tested this assumption by comparing the growth and development of males of the solitary koinobiont endoparasitoid, Cotesia rubecula, in first (L1) to third (L3) larval instars of its preferred host, Pieris rapae and in a less preferred host, Pieris brassicae. Beginning 3 days after parasitism, hosts were dissected daily, and both host and parasitoid dry mass was determined. Using data on parasitoid dry mass, we measured the mean relative growth rate of C. rubecula, and compared the trajectories of larval growth of the parasitoid during the larval and pupal stages using non-linear equations. Parasitoids generally survived better, completed development faster, and grew larger in earlier than in later instars of both host species, and adult wasps emerging from P. rapae were significantly larger than wasps emerging from all corresponding instars of P. brassicae. During their early larval stages, parasitoids grew most slowly in L1 P. rapae, whereas in all other host classes of both host species growth to pupation proceeded fairly uniformly. The growth of both host species was markedly reduced after parasitism compared with controls, with the development of P. brassicae arrested at an earlier stage, and at a smaller body mass, than P. rapae. Our results suggest that C. rubecula regulates certain biochemical processes more effectively in P. rapae than in P. brassicae, in accordance with its own nutritional and physiological requirements. Furthermore, we propose that, for parasitoids such as C. rubecula, which do not consume all host tissues prior to pupation, that parasitoid size and host quality may vary independently of host size at oviposition and at larval parasitoid egression. PMID- 12770387 TI - Effect of food quality on the body temperature of wasps (Paravespula vulgaris). AB - Body surface temperature of individually marked wasps (Paravespula vulgaris, Vespidae, Hymenoptera) was measured by infrared thermography during repeated visits to a feeding bowl without injuring them or disturbing their behavior. Wasps were fed 0.5, 1 and 2 mol/l sucrose solution at two ambient temperatures.Thoracic temperature varied significantly in dependence on food quality (sucrose concentration of solution). At the higher ambient temperatures of 26.1-30.2 degrees C mean thoracic surface temperatures from different experiments were 35.3 and 38.0 degrees C when the wasps took a 0.5 mol/l sucrose solution, 37.0, 38.7 and 38.7 degrees C when they took a 1 mol/l solution, and 39.1 degrees C when they took a 2 mol/l sucrose solution. At the lower ambient temperatures of 17.6-21.0 degrees C thoracic temperatures were lower but the effect of different sucrose concentrations was similar: 34.7 degrees C with a 0.5 mol/l and 36.1 degrees C with a 1 mol/l sucrose solution. The concentration effect amounted to about 10-25% of the whole variability of thorax temperature. By contrast, the temperatures of the head and abdomen did not follow the changes in thorax temperature according to changes in sucrose concentration closely, which suggests that the pattern of haemolymph circulation may have changed after landing, during the wasps' stay at the feeder. At initial landing at the feeders thoracic temperatures where equal to (three of eight tests) or lower (five of eight tests) than at final departure.The correlation of thorax temperature with food quality probably reflects the wasps' level of excitement and motivation to collect the food, which allows them to balance energetic investment with profitability of foraging and the needs of flight muscle performance and motility. PMID- 12770388 TI - The effect of target orientation on the visual acuity and the spatial frequency response of the locust eye. AB - The eye of the locust was shown to distinguish between vertical and horizontal alignments of narrow slit targets, and responded more strongly with horizontal alignments. In all cases the response increased as slit width decreased, to a limit of about 0.1 degrees. Similarly the eye responded more strongly to gratings in horizontal alignment, with a peak in response close to a spatial frequency of 3 cyc/deg. These results are discussed and comparisons are made with previous findings on insect and other eyes. PMID- 12770389 TI - Provision of riboflavin to the host aphid, Acyrthosiphon pisum, by endosymbiotic bacteria, Buchnera. AB - Differential cDNA display and quantitative RT-PCR suggested that the riboflavin synthase complex of the aphid endosymbiont, Buchnera, is active only when the symbiotic system is maintained and well organized in young hosts. Since this finding suggested the provision of riboflavin by Buchnera, we examined the effect of dietary riboflavin on the performance of symbiotic and aposymbiotic aphids using chemically-defined diets. Our results indicate: (1) dietary riboflavin is slightly detrimental to young, symbiotic aphids; (2) dietary riboflavin is essential to aposymbiotic aphids; (3) dietary riboflavin remarkably improves the performance of aposymbiotic aphids. These results strongly suggest that young, symbiotic aphids are provided with riboflavin by their endosymbionts, Buchnera. PMID- 12770390 TI - Performance and host preference of Ascia monuste (Lepidoptera, Pieridae). AB - Experiments were conducted to determine the performance and host preference of Ascia monuste using kale (Brassica oleraceae, var. Acephala) and mustard (B. juncea). These plants differ significantly in water and nitrogen content, with mustard having larger amounts of water and kale larger amounts of nitrogen. The performance results confirmed that kale is a better food source than mustard for the species, even when eggs were collected on mustard leaves in the field. However, when eggs were collected on mustard, the kale nutritive value was lower than the nutritive value obtained when eggs were collected on kale leaves. Furthermore, the results of oviposition preference obtained in the field and in the laboratory have shown a preference for kale, indicating the presence of a positive correlation with performance. In contrast to the data about oviposition preference, there was no immature feeding preference. These results indicate that host selection occurs during the oviposition process. Furthermore, it is possible that the high abundance of kale cultivated in the region studied and the nutritional quality of this plant are two factors that influence the positive relationship between oviposition preference and performance for A. monuste. PMID- 12770391 TI - Recycling of urea associated with the host plant urease in the silkworm larvae, Bombyx mori. AB - Urea concentration and urease activity in the midgut content were compared between larvae of the silkworm, Bombyx mori fed an artificial diet and those fed fresh mulberry leaves. A considerable amount of urea was found in the midgut content of the both larvae, however it was significantly lower in the larvae fed fresh mulberry leaves than in the larvae fed the artificial diet; average urea concentrations in the midgut content of the larvae fed fresh mulberry leaves and the artificial diet were 2.9 and 4.6 &mgr;mol/g, respectively. Urea in the midgut content seems to be secreted from the insect itself since the amount of urea in both diets were negligibly small. Urease activity was detected only in the midgut content of the larvae fed fresh mulberry leaves but not in other tissues of the larvae. On the other hand, no urease activity was detected in the midgut content of the larvae fed the artificial diet. Subsequently, to elucidate the role of mulberry leaf urease in the midgut lumen, larvae that had been reared on the artificial diet were switched to fresh mulberry leaves. The diet switch caused a rapid decrease in urea concentration in the midgut content and an increase in ammonia concentration in the midgut content, suggesting that secreted urea could be hydrolyzed to ammonia by mulberry leaf urease in the midgut lumen. Furthermore, to investigate the physiological significance of mulberry leaf urease on urea metabolism of the silkworm, (15)N-urea was injected into the hemocoel, and after 12 h the larvae were dissected for (15)N analysis. A considerable amount of (15)N was found to be incorporated into the silk-protein of the larvae fed fresh mulberry leaves, but there was little incorporation of (15)N into the silk-protein of the larvae fed the artificial diet. These data indicate that urea is converted into ammonia by the action of mulberry leaf urease in the midgut lumen and used as a nitrogen source in larvae fed mulberry leaves. PMID- 12770392 TI - Effect of high temperature on the metabolic processes affecting sorbitol synthesis in the silverleaf whitefly, Bemisia argentifolii. AB - Whiteflies accumulate the polyhydric alcohol, sorbitol, when exposed to temperatures greater than about 30 degrees C. Feeding experiments using artificial diets containing labeled sucrose showed that more of the label was incorporated into whitefly bodies and less was excreted in the honeydew when feeding was conducted at 41 compared with 25 degrees C. Analysis of the components of the honeydew showed that more of the excreted label was in glucose and fructose and less in trehalulose at 41 degrees C than at 25 degrees C. A similar effect of temperature on honeydew composition occurred for whiteflies feeding on cotton leaves. Measurement of the activities of glycolytic, pentose phosphate and polyol pathway enzymes at 30 and 42 degrees C showed that NADPH dependent ketose reductase/sorbitol dehydrogenase (NADPH-KR/SDH), sucrase, glucokinase and glucose-6-phosphate dehydrogenase activities were stimulated to a greater extent at 42 degrees C than trehalulose synthase and fructokinase. NAD(+) sorbitol dehydrogenase (NAD(+)-SDH) activity was inhibited at 42 degrees C. We propose that high temperature alters metabolic activity in a way that increases the availability of fructose and stimulates pentose-phosphate pathway activity, providing both the substrate and coenzyme for sorbitol synthesis. High temperature also increases the activity of NADPH-KR/SDH, the enzyme in whiteflies that synthesizes sorbitol, but inhibits the activity of NAD(+)-SDH, the enzyme that degrades sorbitol. PMID- 12770393 TI - Nature of the anchors of membrane-bound aminopeptidase, amylase, and trypsin and secretory mechanisms in Spodoptera frugiperda (Lepidoptera) midgut cells. AB - Spodoptera frugiperda larvae have a microvillar aminopeptidase and both soluble and membrane-bound forms of amylase and trypsin. Membrane-bound aminopeptidase is solubilized by glycosyl phosphatidylinositol-specific phospholipase C (GPI-PLC) and detergents, suggesting it has a GPI anchor. Membrane-bound trypsin is not affected by GPI-PLC, although it is solubilized by papain and by different detergents. Membrane-bound amylase is similar to trypsin, although once solubilized in detergent it behaves as a hydrophilic protein. Musca domestica trypsin antiserum cross-reacts with only one polypeptide from S. frugiperda midgut. With this antiserum, trypsin was immunolocalized in the anterior midgut cells at the microvillar surface and on the membranes of secretory vesicles found in the apical cytoplasm and inside the microvilli. The data suggest that in this region trypsin is bound to the secretory vesicle membrane by a hydrophobic anchor. Vesicles migrate through the microvilli and are discharged into the lumen by a pinching-off process. Trypsin is then partly processed to a soluble form and partly, still bound to vesicle membranes, incorporated into the peritrophic membrane. In posterior midgut cells, trypsin immunolabelling is randomly distributed inside the secretory vesicles and at the microvilli surface, suggesting exocytosis. Amylase probably follows a route similar to that described for trypsin in anterior midgut, although membrane-bound forms (peptide anchor) solubilize apparently as a consequence of a pH increase inside the vesicles. PMID- 12770394 TI - The formation of type-I concretions in Drosophila Malpighian tubules studied by electron microscopy and X-ray microanalysis. AB - There are two types of concretions in Drosophila Malpighian tubules: Type-I concretions originate in the distal segments of the anterior tubules, type-II concretions in the adjacent transitional segment between the apical microvilli. Type-I concretions are formed with the aid of carbonic anhydrase within intracellular vesicles, which migrate to the apical cell membrane where they are discharged into the lumen by exocytosis. The carbonic anhydrase inhibitors acetazolamide or hydrochlorothiazide prevent the formation of concretions by interruption of bicarbonate supply. In addition, the formation of concretions can be reduced by feeding with sodium cellulose phosphate. PMID- 12770395 TI - Hormonal control of male horn length dimorphism in the dung beetle Onthophagus taurus (Coleoptera: Scarabaeidae). AB - Male dung beetles (Onthophagus taurus) facultatively produce a pair of horns that extend from the base of the head: males growing larger than a threshold body size develop long horns, whereas males that do not achieve this size grow only rudimentary horns or no horns at all. Here we characterize the postembryonic development of these beetles, and begin to explore the hormonal regulation of horn growth. Using radioimmune assays to compare the ecdysteroid titers of horned males, hornless males, and females, we identify a small pulse of ecdysteroid which is present in both hornless males and females, but not in horned males. In addition, we identify a brief period near the end of the final (third) larval instar when topical applications of the juvenile hormone analog methoprene can switch the morphology of developing males. Small, normally hornless, males receiving methoprene during this sensitive period were induced to produce horns in 80% of the cases. We summarize this information in two models for the hormonal control of male dimorphism in horn length. PMID- 12770396 TI - Water loss and metabolic water in starving Argas reflexus nymphs (Acari: Argasidae). AB - Basic components of the water balance were determined in A. reflexus unfed second instar nymphs kept at 30 degrees C and different relative humidities in darkness. At 2.5% and 32.5% RH, A. reflexus survived for 170+/-60 and 246+/-71 days, respectively. At 56%, 75.5% and 96.5% RH, the experiment was terminated after 168 days without any mortality. The initial water content of hydrated A. reflexus kept at 85% RH was 70.3+/-1.8% while the water content was close to 50% in ticks dehydrated at /=56% suggesting that A. reflexus increased its oxidative metabolism under dehydrating conditions and thus delayed lethal dehydration. Interestingly, the dry mass of dead ticks was almost identical irrespective of whether the ticks had previously been kept at 2.5% and 32.5% RH, i.e. irrespective of their different survival periods. This raises the question whether the ticks had died by dehydration or by exhaustion of energy reserves. PMID- 12770397 TI - The 'Inka cell' and its associated cells: ultrastructure of the epitracheal glands in the gypsy moth, Lymantria dispar. AB - The epitracheal glands in pharate and young pupae of Lymantria dispar are located at the base of ventrolateral tracheal trunks in the prothoracic and first through eighth abdominal segments. Each gland is composed of four cells the ultrastructure of which is described in this paper. One large cell and one smaller cell have an endocrine function, while a third cell is exocrine. A fourth cell forms a canal running from the exocrine cell into the trachea. The large endocrine cell, but not the smaller endocrine cell has released its secretions in freshly moulted pupae. The exocrine cell is assumed to be involved in the pupal moult events as well. The physiological role of the different cell types is discussed: The large endocrine cell (type I endocrine cell) is supposedly homologous with the 'Inka cell', which produces ecdysis triggering hormone (ETH) and was previously described in Manduca sexta; the functions of the smaller endocrine cell (type II endocrine cell) and the exocrine cell remain unknown. PMID- 12770398 TI - Eicosanoids mediate nodulation reactions to bacterial infections in adults of the cricket, Gryllus assimilis. AB - Nodulation is the temporally and quantitatively most important cellular defense reaction to bacterial infections in insects. Inhibition of eicosanoid biosynthesis in adults of the cricket, Gryllus assimilis, immediately prior to intrahemocoelic injections of the bacterium, Serratia marcescens, sharply reduced the nodulation response. Separate treatments with specific inhibitors of phospholipase A(2), cyclooxygenase, and lipoxygenase reduced nodulation, supporting our view that nodule formation is a complex process involving lipoxygenase and cyclooxygenase products. The inhibitory influence of dexamethasone was apparent within 2h of injection, and nodulation was significantly reduced, relative to control crickets, over 22h. The dexamethasone effects were reversed by treating bacteria-injected insects with the eicosanoid precursor polyunsaturated fatty acid, arachidonic acid. Low levels of arachidonic acid were detected in fat body phospholipids, and fat body preparations were shown to be competent to biosynthesize eicosanoids from exogenous radioactive arachidonic acid. These findings in a hemimetabolous insect broaden our hypothesis that eicosanoids mediate cellular immune reactions to bacterial infections in most, if not all, insects. PMID- 12770399 TI - Electrophysiological responses of antennal receptor neurons in female Australian sheep blowflies, Lucilia cuprina, to host odours. AB - Studies were conducted to investigate the electrophysiological responses of receptor neurons on the antennae of female Lucilia cuprina to putative stimulants. Three distinct types of olfactory neurons were identified. Tests with 25 compounds showed 27 out of the 75 neurons examined responded best to racemic 1 octen-3-ol (Oct-best neuron), nine to dimethyldisulphide (DMDS-best neuron) and seven to 2-phenylethanol (PE-best neuron). The remaining cells were not stimulated by any chemical stimulus presented in this study. The olfactory neurons showed low spontaneous activities, 2.2 ~ 2.7 impulses/s, which increased significantly on stimulation with test compounds. Dose-dependent responses were observed with Oct-best neurons. Another type of neuron that had a much higher spontaneous activity (26 impulses/s) was observed. This neuron did not respond to any of the candidate compounds tested but showed a decrease in activity with increased airflow over the antennae and was termed the Airflow-sensitive neuron. The spontaneous activities of Oct-, DMDS-, PE-best and Airflow-sensitive neurons were inhibited by butanoic, pentanoic, hexanoic acids, and 3-methylindole. PMID- 12770400 TI - Enhancement of short wing formation and ovarian growth in the genetically defined macropterous strain of the brown planthopper, Nilaparvata lugens. AB - When JH II, III or methoprene was applied in the nymphal stages to two different strains of the brown planthopper which were selected to produce long (macropterous) or short (brachypterous) wing forms, no effect was observed on the molting profile or metamorphosis. Brachypterization of a majority of the presumptive macropters was, however, observed by application of these chemicals, although there was no effect on wing form in the presumptive brachypters. The results show that the sensitive periods for the brachypterization of the presumptive macropters falls between early antepenultimate instar and within 1 or 2days of the penultimate instar, and that the chemicals were effective, in the following order of potency: methoprene>JH III>JH II. Ovarian growth was greatly enhanced in the presumptive macropters when JH III or methoprene was applied twice, within 12h of the 3rd or 4th nymphal instar and 6h before adult emergence. JH II on the other hand had no effect on ovarian growth when applied to the presumptive macropters at any of the nymphal stages. None of the chemicals had any effect on ovarian growth in the presumptive brachypters. PMID- 12770401 TI - Morphology and ultrastructure of sense organs on the ovipositor of Trybliographa rapae, a parasitoid of the cabbage root fly. AB - The ovipositor of the parasitoid wasp Trybliographa rapae was examined by scanning and transmission electron microscopy. Characteristic peg-like sensilla with a cuticular ring at the base are found at the tip of the ventral valves, where they occur in a characteristic arrangement of triplets. The unusual basal structure probably protects the sensilla against damage during movement through the substrate and piercing of the host cuticle. These sensilla are each innervated by six dendrites, some of which have lamellated tips, generally considered to be characteristic of thermosensitivity. It is suggested that the remaining dendrites are gustatory, and as such probably respond to factors present in host haemolymph. A second type of peg-like sensillum is found on both the dorsal and the ventral valves. These are set in deep pits so that only the tip of the peg protrudes above the surface of the cuticle. These occur along the length of the ovipositor shaft and ultrastructural studies reveal the pegs to be innervated by a single mechanosensitive dendrite, probably monitoring the movement of the ovipositor through the substrate. PMID- 12770402 TI - A factor causing stable stimulation of juvenile hormone synthesis by Diploptera punctata corpora allata in vitro. AB - Co-incubation of corpora allata (CA) from the cockroach, Diploptera punctata, with ovaries, fat body or muscle but not brain or testis, leads to a substantial increase in juvenile hormone synthesis. Incubation of the glands in medium pre conditioned with ovaries also stimulates JH synthesis. The ovary was used as a convenient source of stimulatory factor for a detailed analysis of its physiological effects on the CA. The increase in JH synthesis is stable, maintained over 24h after exposure to the stimulatory factor. Stimulation is dose dependent, and the corpora allata show an exquisite relationship between sensitivity to this factor and developmental stage. Day 0 and day 1 glands, as well as glands from post-vitellogenic females, are sensitive to stimulation, whereas glands from vitellogenic females are not sensitive. Corpora allata attached to the brain do not respond to the stimulatory factor, and denervation in vivo leads to an increase in JH synthesis by the glands and a loss in sensitivity to the factor. These data suggest that glands from pre- and post vitellogenic females are inhibited by their nervous connection to the brain. In contrast, glands from vitellogenic females are normally responding to the endogenous stimulatory factor and are thus no longer stimulated in vitro. Co incubation of CA with allatostatin and conditioned medium still leads to a stimulation of JH synthesis, suggesting that the restraining effect of the nervous connections to the brain is not caused by allatostatin. The CA cell number increases between emergence and day 2, then remains stable until after oviposition. The stimulatory factor accelerates the increase in cell number in young adult females. The results are interpreted as providing evidence for a constitutive change in CA activity caused by a humoral factor produced by various tissues including the ovary, and modulated by nervous connections to the brain. PMID- 12770403 TI - Parallel control mechanisms underlying locomotor activity and sexual receptivity of the female German cockroach, Blattella germanica (L.). AB - We have studied the effects of ovaries, juvenile hormone (JH) and mating on locomotor activity and sexual receptivity of female German cockroaches. Our results indicate that locomotor activity and sexual receptivity are under the same control mechanisms. The ovary served as a negative masking factor for the locomotor circadian rhythm, but did not affect the frequency of locomotor activity. We conclude that JH controls the locomotor activity of females from the following evidence: (1) increasing locomotion of virgin females coincided with an increasing volume of the corpora allata; (2) allatectomy reduced female locomotion significantly; (3) after absorbing the JH analogue (fenoxycarb) through their tarsi, allatectomized females regained their high level of locomotor activity. Since the daily locomotor activity of allatectomized and ovari-allatectomized females changed cyclically with continuous (non-cyclic) contact of fenoxycarb, an unidentified factor which was independent of ovarian development is proposed to regulate cyclic locomotor activity. In addition to controlling the frequency of locomotor activity, JH was essential for the expression of the locomotor circadian rhythm because allatectomy abolished the circadian rhythm expressed in ovariectomized females. Mating significantly decreased the frequency of locomotor activity and the degree of sexual receptivity. The inhibitory effect of mating resulted from the transmission of a mating signal through the ventral nerve cord when sperm was transferred successfully. The mating experiments with allatectomized and ovariectomized females showed that JH was the major factor in regulating the expression of sexual receptivity. PMID- 12770404 TI - Origin and pathway of the epidermal secretion in the damselfly head-arresting system (Insecta: Odonata). AB - In damselflies, the arrester system is responsible for an additional attachment of the head to the neck. It consists of a pair of mobile postcervical sclerites (SPC) covered by microtrichia. In their lateral position, SPCs can fixate the head on fields of microtrichia on the back surface of the head. The intact surface of microtrichia of the SPC is usually covered by a lipid-containing secretion. The present study provides ultrastructural data on the secretory epidermis and pore channels adapted to transport the secretion to the cuticle surface. 1) Shock-frozen preparations of the contact area show that microtrichia of co-opted surfaces do not interlock with each other. When co-opted fields are pressed to each other, deformations of flexible microtrichia of the SPC result in an increase of contact area between corresponding structures and consequently in an increase of frictional forces. 2) In the area of the SPC, only electron-lucent vesicles have been found. Histochemical procedures revealed that the material stored in vesicles and liberated on the external surface of the SPC is presumably non-volatile lipid. 3) Only a small number of large pore channels reach the surface of a microtrichium. Most of them terminate with tiny terminal channels, whose diameter is approximately six to twenty times smaller than the diameter of structures of secretory blooms occurring in SEM in shock-frozen and air-dried preparations. The terminal channels do not penetrate the epicuticular layer. It seems that the secretion reaches the epicuticle through terminal channels and diffuses through the epicuticle without any channel structures. PMID- 12770405 TI - Arguments for two distinct gonadotropic activities triggered by different domains of the ovary maturating parsin of Locusta migratoria. AB - To complete previous results concerning the role of the ovary maturating parsin of Locusta migratoria (Lom OMP), we determined, by an enzyme immunoassay, the titers of circulating ecdysteroids and analyzed circulating vitellogenin (Vg) and oocyte growth following (1) suppression of 20 hydroxyecdysone (20E) and (2) injection of the Lom OMP, either as an entire molecule in allatectomized adults or as smaller peptides in allatectomized fifth-instar larvae females. Titers of ecdysteroids appeared unrelated to the presence of circulating Vg but increased during the first phase of vitellogenesis and injection of OMP accelerated the occurrence of circulating 20E. Nevertheless, immunoneutralization of 20E at the beginning of adult life delayed but did not prevent rapid oocyte growth contrary to immunoneutralization of Lom OMP suggesting an additive gonadotropic effect of the neurohormone, distinct from that of 20E. Of two synthetic peptides corresponding to the C- and N-terminal gonadotropic domains of the OMP, respectively, only the C-terminal peptide was able to induce Vg in allatectomized larvae. After metamorphosis, injection of OMP did not induce Vg in adults allatectomized at the beginning of imaginal life but improved the maintenance of circulating Vg in adults allatectomized after Vg appeared in the haemolymph. This result suggests that OMP either delays the Vg mRNA decay or increases the translation of Vg mRNA. Thus, Lom OMP appears to have two distinct roles: an ecdysteroidogenic effect triggered by its C-terminal domain with the ovary as the target tissue and a protecting effect on Vg mRNA probably triggered by its other gonadotropic domain, the N-terminal, with the fat body as the target tissue. PMID- 12770406 TI - Dinocampus (=Perilitus) coccinellae teratocyte-specific polypeptide: its accumulative property, localization and characterization. AB - Dinocampus (=Perilitus) coccinellae (Braconidae: Hymenoptera) teratocytes synthesize a teratocyte-specific polypeptide (TSP) with a high molecular weight of 540kDa. The TSP has a tendency to accumulate in the teratocyte cells without release after synthesis ([Okuda and Kadono-Okuda, 1995]), which was confirmed in this study. Pulse-chase fluorography indicated that teratocytes at a younger stage (6 days after parasitization)secreted negligible TSP into the medium after synthesis, while teratocytes at an older stage (11 days after parasitization)secreted the synthesized products into the medium, although the amount released was still low. Western blot with anti-TSP serum showed that only a small amount of TSP appeared in the parasitized host hemolymph, even when TSP synthesis by teratocytes was actively taking place, which also supported the accumulative nature of TSP. The immunoelectronmicroscopic studies revealed that the TSP was localized specifically in high electron-dense vacuoles. Lectin blot analysis identified TSP as a high mannose glycoprotein. The amino acid composition of the major subunit of the TSP was quite similar to that of nutritive proteins such as vitellogenin and storage proteins of some insects. These characterization data, together with the accumulation property of the TSP indicates that Dinocampus teratocyte primarily plays a nutritive role for the developing parasitoid larvae. TSP exhibited esterase activity, which indicates that TSP may have an additional function in the host-parasitoid reaction. PMID- 12770407 TI - Body size and cell size in Drosophila: the developmental response to temperature. AB - In Drosophila, like most ectotherms, development at low temperature reduces growth rate but increases final adult size. Cultures were shifted from 25 degrees C to low (16.5 degrees C) or to high (29 degrees C) temperature at regular intervals through larval and pupal stages, and the flies of both sexes showed an increase or decrease, respectively, in the size of thorax, wing and abdominal tergite. Size changes in the wing blade resulted from changes in the size of the epidermal cells (with only a small increase in cell number in males reared at low temperature). The temperature-shifts became less effective as they were made at successively later developmental stages, demonstrating a cumulative effect of temperature on adult size. The thorax and wing develop from the same imaginal disc, with most cell division occurring in larval stages, but they differ in timing of temperature sensitivity, which extends only to pupariation or into the late pupal stage, respectively. Growth of the adult abdomen occurs largely after pupariation but its size is temperature-sensitive through both larval and pupal stages. We discuss growth control in Drosophila and the likely effects of temperature on food assimilation, growth efficiency and allocation of nutrients to the production of different tissues. PMID- 12770408 TI - Hsp70 and larval thermotolerance in Drosophila melanogaster: how much is enough and when is more too much? AB - Heat shock proteins (Hsps) and other molecular chaperones perform diverse cellular roles (e.g., inducible thermotolerance) whose functional consequences are concentration dependent. We manipulated Hsp70 concentration quantitatively in intact larvae of Drosophila melanogaster to examine its effect on survival, developmental time and tissue damage after heat shock. Larvae of an extra-copy strain, which has 22 hsp70 copies, produced Hsp70 more rapidly and to higher concentrations than larvae of a control strain, which has the wild-type 10 copies of the gene. Increasing the magnitude and duration of pretreatment increased Hsp70 concentrations, improved tolerance of more severe stress, and reduced delays in development. Pretreatment, however, did not protect against acute tissue damage. For larvae provided a brief or mild intensity pretreatment, faster expression of Hsp70 in the extra-copy strain improved survival to adult and reduced tissue damage 21h after heat shock. Negative effects on survival ensued in extra-copy larvae pretreated most intensely, but their overexpression of Hsp70 did not increase tissue damage. Because rapid expression to yield a low Hsp70 concentration benefits larvae but overexpression harms them, natural selection may balance benefits and costs of high and low expression levels in natural populations. PMID- 12770409 TI - Adult diapause and cold hardiness in Aulacophora nigripennis (Coleoptera: Chrysomelidae). AB - We investigated the control of diapause termination and seasonal changes of cold hardiness and polyol content in Aulacophora nigripennis. Adults were ready to start post-diapause development upon transfer to high temperature by late February irrespective of photoperiod. Photoperiod probably functions to maintain diapause before winter because adults resume reproductive development at a long photoperiod in autumn. Adults showed a decreased supercooling point (SCP), increased chill tolerance and high myo-inositol content during winter. Chill tolerance at 0 degrees C appears to be a more suitable indicator of their cold hardiness than SCP because they die at 0 degrees C without freezing and they normally have no chance of being exposed to low subzero temperatures close to their SCP. The temporal pattern for changes in chill tolerance was synchronized with that for fluctuations in myo-inositol content, indicating a possible causal relationship between the two phenomena. PMID- 12770410 TI - Temporal profiles of juvenile hormone titers and egg production in virgin and mated females of Heliothis virescens (Noctuidae). AB - Juvenile hormones (JH) I, II, and III were monitored in hemolymph of virgin and mated females of various ages in Heliothis virescens. JH I was the predominant homologue followed by JH II, but JH II was present at a higher level in young virgin females. JH III was detectable only at a low level. In virgin females, hemolymph JH titers were low at emergence (2.2ng/ml-total amount of JH I and JH II), but increased thereafter and reached a maximum at 24h of age (53.5ng/ml). At 30h and 36h of age, JH titers dropped to a low level, but increased again in older virgin females. After mating, JH titers increased significantly. JH titers at 0h after uncoupling (137.4ng/ml) were nearly 3 times as high as those in 24-h old virgin females. Within 6h after uncoupling, JH titers decreased slightly, but titers increased with age of mated females and reached a level of 320.2ng/ml hemolymph at 72h after uncoupling. The titer of JH I and JH II was correlated highly with total number of eggs produced (r(2)=0.70, P<0.001). Mating stimulated JH production, resulting in an increase in egg production. PMID- 12770411 TI - Evidence for the participation of arachidonic acid metabolites in trehalose efflux from the hormone activated fat body of the cockroach (Periplaneta americana). AB - The hypertrehalosemic hormones, HTH-I and HTH-II, activate trehalose synthesis and increase the rate of sugar efflux from Periplaneta americana fat body in vitro. These processes are unaffected by the diacylglycerol, 1-oleyl-2-acetyl-sn glycerol, an activator of protein kinase C. Similarly, H-7 and spingosine, inhibitors of protein kinase C, are also inactive against trehalose efflux. The possibility that diacylglycerol lipase might generate an active fatty acid species was ruled out because of the failure of the inhibitor RHC-80267 to inhibit trehalose efflux. Activation of trehalose efflux from the intact fat body by HTH-I was strongly inhibited in a concentration dependent manner by the cyclooxygenase inhibitors indomethacin and diclofenac, but not by acetylsalicylic acid. Nordihydroguaiaretic acid, a lipoxygenase inhibitor, also blocked HTH-I activated trehalose efflux in a concentration dependent fashion. The phospholipase A(2) inhibitors mepacrine and 4'-bromophenacyl bromide were also effective in decreasing the efflux of trehalose from HTH-I challenged fat body. The data suggest possible roles for arachidonic acid metabolites in the regulation of trehalose synthesis and in the efflux of the sugar from the fat body. PMID- 12770412 TI - The effects of reproduction on longevity and fertility in male Drosophila melanogaster. AB - We examined the effect of reproduction on subsequent survival and fecundity of male Drosophila melanogaster by reversing the reproductive status of individuals part-way through life. Reproduction had a much more marked effect on fertility than survival: males with a history of reproduction showed complete sterility at a time when upwards of 80% of their cohort were still alive. Analyses of survival rates alone gave a misleading measure of the impact of ageing. Sterility appeared to be attributable mainly to a reduction in sperm count. Early reproduction caused permanent, irreversible damage to both survival and fecundity, with risk playing an apparently minor role. Individual differences in frailty appeared to be of little consequence for the interpretation of these reversal experiments, although its possible occurrence made definite detection of risk difficult. PMID- 12770413 TI - Immune responses in Rhodnius prolixus: influence of nutrition and ecdysone. AB - Starved larvae of Rhodnius prolixus, when challenged with Enterobacter cloacae B12, had their mortality related to their period of starvation. R. prolixus larvae fed on plasma alone, compared with insects fed on whole blood, had their immune reactivity affected as shown by: (i) a significant reduction in the ability to produce cecropin-like and lysozyme activities in the haemolymph when inoculated with E. cloacae; (ii) a reduction in numbers of haemocytes and nodule formation following challenge with bacteria; (iii) a decreased ability of plasma fed insects in destroying their infection caused by inoculation of E. cloacae cells; and (iv) alpha-ecdysone therapy counteracted the immune depression in Rhodnius larvae fed on plasma alone. However, unlike other immune reactions, this set of experiments failed to demonstrate any interference of the plasma feeding on the prophenoloxidase-activating system, since melanin production was not reduced when the system was stimulated by the presence of bacteria in the haemolymph. The significance of these data is discussed in relation to the effect of diet components and the moulting hormone on the immune reactivity in insects. PMID- 12770414 TI - Characterization and temporal aspects of haemolymph juvenile hormone esterase in adult cockroach, Periplaneta americana. AB - Daily variations in the in vitro haemolymph juvenile hormone esterase activity (hJHE) of adult male and female Periplaneta americana were monitored over a 2 week period from the time of adult emergence and throughout the first reproductive cycle of the adult female. Kinetic analysis of hJHE from females indicated an apparent K(m) JH III of 5.59+/-1.75&mgr;M (V(max)=140pmol JH III hydrolysedh(-1) per &mgr;l haemolymph). In females the mean rate of JH III metabolism in diluted haemolymph shortly after emergence was 27.5+/-1.5pmolh( 1)&mgr;l(-1) (n=16) and remained relatively low (16-32pmolh(-1)&mgr;l(-1)) over much of early adult development. Activity remained at this level during the first two days of the 4 day reproductive cycle, but showed a much increased broad peak of activity (74-93pmolh(-1)&mgr;l(-1)) at 60-72h post-extrusion. This peak lags behind the whole body JH titre peak, which could suggest that elevated levels of JH III may bring about the induction of JH esterase(s). A different pattern of JHE activity was observed in adult males. hJHE rates in males at emergence were almost twice as high (81.5+/-15.8pmolh(-1)&mgr;l(-1), n=16) as those found in females at this time, but thereafter showed a downturn to moderate levels (44 68pmolh(-1)&mgr;l(-1)) that were maintained for the remainder of the study. Rapid (FPLC) DEAE-sepharose ion exchange chromatography, ultrafiltration and fast-flow superose gel filtration chromatography were employed to achieve an efficient partial purification (166-fold) of the hJHE from cell-free plasma of reproductively active female P. americana 48-72h post-ootheca extrusion. Gel filtration and SDS-polyacrylamide gel electrophoresis (PAGE) revealed an enzyme having apparent molecular mass of between 60 and 70kDa, whilst non-denaturing PAGE and iso-electrofocusing resolved a single acidic enzyme peak with a pI of 4.9. PMID- 12770415 TI - Control of pupal fat body disappearance in the female black blow fly, Phormia regina (Meigen) by the brain and the corpus allatum. AB - Neck-ligation, brain implantation, allatectomy, methoprene treatment, and ovariectomy indicated that the disappearance of pupal fat body cells in newly emerged adult female blow fly, Phormia regina, is controlled by the brain and the corpus allatum (CA). Absence of brain or CA greatly lowered the rate of fat body cell disappearance (i.e. death). Dependency on the CA decreased from 0 to 36h post-emregence, indicating that the CA was active during the earlier part of this timespan. Methoprene treatment enhanced pupal fat body cell disappearance in allatectomized females. Brain implantation restored the rate of pupal fat body cell disappearance in neck-ligated flies. Brains from day 1 sugar-fed flies proved to be more effective than those from day 2 sugar-fed flies, indicating that there may be a window after adult emergence that allows the brain to act directly or indirectly on the death of pupal fat body cells. Ovariectomy did not alter the rate of pupal fat body cell death in test animals. Dying pupal fat body cells were smaller in size, less dense (i.e. did not sink in saline like normal pupal fat body cells), and stickier (i.e. attached to other tissues tighter) than the healthy cells. A possible role played by ecdysteroids is also discussed. PMID- 12770416 TI - Relationship of the neutral sterols and ecdysteroids of the parasitic mite, Varroa jacobsoni to those of the honey bee, Apis mellifera. AB - The neutral sterols of the parasitic mite Varroa jacobsoni were compared with Apis mellifera carnica drone pupae. Analysis by GLC-mass spectrometry indicated mite sterols were reflective of the sterol composition of the drones; 24 methylenecholesterol was the major sterol in both species, with lesser amounts of sitosterol and isofucosterol. Cholesterol accounted for less than 1% of the total sterols. Ecdysteroid analyses indicated drones contained primarily makisterone A. In addition to makisterone A, mites contained ecdysone and 20-hydroxyecdysone, which accounted for over 66% of the ecdysteroid detected. These results indicate that while V. jacobsoni are apparently unable to convert dietary sterols to cholesterol, they are able to produce significant amount of C(27) ecdysteroids in a low cholesterol environment. PMID- 12770417 TI - Impact of feeding, molting and relative humidity on cuticular wax deposition and water loss in the lone star tick, Amblyomma americanum. AB - To determine whether there is a correlation between the amount of lipids on the surface of ticks and their permeability to water, we quantified cuticular surface wax and measured water-loss rates in the ixodid tick Amblyomma americanum from nonfed nymph to egg-laying female. Ticks deposited no extra cuticular lipids during feeding, permitting maximum transpiratory water loss that presumably helps to concentrate the bloodmeal; and ticks deposited additional cuticular wax after apolysis that reduced integumental water loss, which likely prepares ticks for off-host existence. A remarkable three-fold boost in surface wax deposition and extreme water retention were noted after host drop-off following feeding. This wax is likely host-derived. Fed nymphs could discriminate between low and high relative humidity, enabling pharate adults to conserve lipid that would otherwise be lost with the exuvia and feces. This conservation strategy likely adds to the lipid pool needed by the tick to survive in a dry environment and complements the tick's behavioral abilities for seeking out optimum conditions for water conservation and host location. PMID- 12770418 TI - Caste formation in the polyembryonic wasp Copidosoma floridanum (Hymenoptera: Encyrtidae): in vivo and in vitro analysis. AB - The polyembryonic wasp Copidosoma floridanum produces two morphologically distinct types of larvae in its host Trichoplusia ni. Reproductive larvae consume the host, pupate, and form adult wasps, whereas precocious larvae manipulate the sex ratios of the reproductive caste and defend the brood against interspecific competitors. The previous study indicated that morphogenesis of the reproductive caste was associated with a 9-day competency period, and that ecdysteroids of host origin were required for completion of embryogenesis. Here we investigated whether factors associated with the host environment mediate morphogenesis of precocious larvae and caste determination. Embryogenesis of precocious larvae was found to be synchronized with specific stages of the host first-fourth instars. However, development of precocious larvae did not depend on environmental factors specifically associated with these host stages. Elevation of the host juvenoid titer using the analogue methoprene induced T. ni to undergo a supernumerary sixth instar, but did not alter the proportion of wasp embryos that developed into precocious and reproductive larvae. In contrast, embryos competent to initiate morphogenesis developed into precocious larvae when transplanted into novel host stages such as pupae. Development of precocious larvae was arrested by ablation of the host's source of ecdysteroids, but could be rescued dose dependently by injection of 20-hydroxyecdysone. In vitro rearing studies confirmed that completion of embryogenesis of the precocious caste required an exogenous pulse of 20-hydroxyecdysone. Combined with previous studies, our results indicate that embryos forming precocious and reproductive larvae acquire the competence to undergo morphogenesis at different times. However, we find no evidence to suggest that caste determination is mediated by environmental factors associated with a specific stage of the host. PMID- 12770419 TI - Effects of photoperiod and aging on locomotor activity rhythms in the onion fly, Delia antiqua. AB - At photoperiods longer than 8h per 24h, adults of the day-active onion fly Delia antiqua showed a major peak of locomotor activity in the late photophase and also bursts of activity induced by lights-on or lights-off. At shorter photoperiods the activity peaks fused. After transfer from long photoperiods to constant darkness (DD), the rhythm free-ran, but only the major peak persisted. This suggests that only the major peak is controlled by the circadian pacemaker. At long photoperiods, the daily phase of the major peak occurred progressively later with age. As a result, the activity at short photoperiods often shifted from photophase to scotophase in old flies. The free-running period (tau) also changed with age; tau was shorter than 24h until 14-20 days after eclosion and thereafter became longer, but a few individuals repeated changes in tau. The phase delay of locomotor activity with age in D. antiqua would be attributable to the increase in tau. PMID- 12770420 TI - Effects of Microplitis croceipes Teratocytes on Host Haemolymph Protein Content and Fat Body Proliferation. AB - Qualitative and quantitative changes in haemolymph proteins in Heliothis virescens were observed in larvae injected with either Microplitis croceipes teratocytes or teratocyte secreted proteins (TSP). Haemolymph protein titres in hosts receiving either 0.5 or 1 larval equivalent (LE) of teratocytes were similar to those of parasitized larvae, whereas a single injection of 4LE of TSP was required to induce a similar response. SDS-PAGE showed that the 82kDa monomer of riboflavin-binding protein and the 74/76kDa monomers of storage proteins were significantly reduced in parasitized larvae and in nonparasitized larvae treated with TSP. Concentrations of a 155kDa monomer (insectacyanin chromoprotein) also were reduced in parasitized larvae and those injected with either teratocytes or TSP. Two monomers (56 and 60kDa) were unique to parasitized larvae. Treated larvae required several days longer than controls to reach a comparable premetamorphic stage (burrowing-digging). Reductions in fat body proliferation similar to those seen in parasitized larvae were observed in larvae treated with either 1LE of teratocytes, or with 2 or 4LEs of TSP. Perivisceral fat body weights from larvae treated with either 0.25 or 0.5LE of teratocytes were significantly reduced, but less so than those which received 1LE. Thus, fat body proliferation in both teratocyte- and TSP-treated larvae was inhibited in a dose dependent manner. Both light- and transmission electron microscopy observations revealed cytological differences in fat body tissues of larvae injected with either teratocytes or TSP from the condition observed in parasitized larvae and noninjected controls. Gross dissection of periviseral fat body from parasitized, teratocyte-injected and TSP-injected larvae showed tissue much less developed and differing considerably in appearance from controls. Observed differences included reduced size and/or number of lipid bodies and qualitative and quantitative changes in other cytoplasmic organelles. PMID- 12770421 TI - Effect of temperature on cold-hardiness and tissue ice formation in the adult chrysomelid beetle Melasoma collaris L. AB - The freeze-tolerant chrysomelid beetle Melasoma collaris overwinters in plant litter on windswept ridges or covered with snow for 8-9 months in the Norwegian alpine region. Lower lethal temperature, supercooling and melting point depression were correlated to accumulation of glycerol. The lower limit of freeze tolerance was associated with the freezing of 73-75% body water. About 23-15.5% of the body water was osmotically inactive, and the highest percentage was revealed in individuals depleted of glycerol at 21 degrees C. A shift in cooling rate from 1 degrees Cmin(-1) to 1 degrees C every 13.5min lowered nucleating temperature markedly. The alteration in nucleating activity probably arises from the structure of the haemolymph nucleating agent that functions to slow embryo growth at the slow cooling rate. An enhanced supercooling is particularly beneficial in autumn before M. collaris has accumulated glycerol, since supercooled individuals accumulate glycerol in higher concentrations than frozen ones. Freezing at higher temperatures is probably a better survival strategy during brief intervals with pronounced decrease in air temperature. PMID- 12770422 TI - Second messenger-controlled membrane conductance in locust (Locusta migratoria) olfactory neurons. AB - The membrane conductance of olfactory neurons of Locusta migratoria was examined using the whole-cell configuration of the patch-clamp technique. Intracellular application of the second messenger inositol 1,4,5 trisphosphate (IP(3)) via a dual pipette technique elicited a clear increase in the membrane conductance. The IP(3)-induced conductance increased due to a rise in the extracellular concentration of calcium from 100&mgr;M to 4mM. Micromolar concentrations of ruthenium red partially blocked the IP(3)-induced increase in membrane conductance. Stimulating olfactory neurons with odour (hexenoic acid) resulted in an increase in the membrane conductance partially similar to that mediated IP(3). These findings suggest that stimulation with appropriate odours as well as intracellular application of IP(3) activate the same calcium-permeable ion channels in the plasma membrane of insect olfactory neurons. PMID- 12770424 TI - Effects of sensory stimuli on the behavioural phase state of the desert locust, Schistocerca gregaria. AB - The nature of stimuli, emanating from other locusts, which are effective in inducing gregarization in the desert locust was investigated. Isolated-reared fifth-instar nymphs were subjected to tactile, visual and olfactory stimuli, presented singly and in combination, and the effect on the behavioural phase state was quantified using logistic regression analysis. Tactile stimulation provided by rolling paper spheres proved to be highly gregarizing, whether presented alone or in combination with the other stimuli. Olfactory and visual stimuli together caused partial behavioural gregarization. Visual stimulation alone was weakly gregarizing after prolonged exposure, while olfactory stimuli alone were ineffective. Nymphs and pre-reproductive and reproductive adults of both sexes were also treated with synthetic adult male 'aggregation' pheromone blend (Torto et al., 1994, Journal of Chemical Ecology 20, 1749). No effect of this blend was found on the behavioural phase state, even when visual stimuli were present. Non-locust related stimuli, including wheat odour and flashing lights, were also tested on nymphs. Neither induced any change in the behavioural phase state, indicating that increased sensory flow is not a sufficient explanation for locust-induced behavioural phase change. PMID- 12770423 TI - Hemolymph ecdysteroid titer and ecdysteroid-dependent developmental events in the last-larval stadium of the silkworm, Bombyx mori: role of low ecdysteroid titer in larval-pupal metamorphosis and a reappraisal of the head critical period. AB - The endocrine regulation of larval-pupal metamorphosis was studied in the silkworm, Bombyx mori, by measuring the following changes: hemolymph ecdysteroid titer, the secretory activity of prothoracic glands and the responsiveness of larvae to ecdysteroids and prothoracicotropic hormone (PTTH), with regard to developmental events such as the occurrence of spinneret pigmentation, initiation of cocoon spinning and onset of wandering stage as indicated by gut purge. These measurements were concentrated especially on the time before and after the head critical period (HCP) which falls 3-4 days before the gut purge ([Sakurai, 1984]). A small increase in the hemolymph ecdysteroid titer was first found during the HCP, and then the titer increased with daily fluctuations. Small but significant titer peaks were found prior to the occurrence of both spinneret pigmentation and gut purge, indicating that an individual titer peak could possess a specific role in development. Responsiveness of larvae to exogenous 20 hydroxyecdysone (20E) after the HCP was markedly higher than that before the HCP. The sensitivity of the prothoracic gland to PTTH also changed during the HCP. The results thus showed that the HCP is not the period after which an additional PTTH release is not required for the developmental events occurring on schedule, but rather it is the period during which complex events occur not only in the endocrine glands but also in the peripheral tissues. In addition, various developmental phenomena before gut purge are brought about by the hemolymph ecdysteroid whose concentration gradually increased with daily fluctuations, and these precise changes in the titer appeared to be important for the sequential occurrence of developmental events in the larval-pupal metamorphosis. PMID- 12770425 TI - Measurement of age and population age structure in the blowfly, Lucilia sericata (Meigen) (Diptera: Calliphoridae). AB - For field samples of the blowfly Lucilia sericata (Meigen), the degree of wing abrasion, corrected for individual size, was shown to give a useful estimate of age, by calibration against a known measure of chronological age, the fluorescence of pteridines in the head capsule. In more extensive field studies, therefore, wing fray was used to indicate the likely ovarian cycle of each female, after which the length of the most advanced egg follicle was used to give a more precise estimate of age within this cycle. Using these two complementary techniques, the ages of 806 female L. sericata, from two farms in the south-west of England, were estimated over a complete field season in 1996. Analysis of median age of these females revealed distinct peaks and troughs over time. The observed periods of low median age corresponded with predicted periods of high adult emergence; the periods of increasing and high median age corresponded with the predicted senescence of the L. sericata population. Hence, by analysis of population age structure, this study supports the view that fluctuations in abundance of L. sericata, observed in the field, represent the emergence and senescence of relatively discrete generations of adults. PMID- 12770426 TI - Spermatophore formation and regulation of egg maturation and oviposition in female Heliothis virescens by the male. AB - This study was undertaken, using various surgical manipulations, to examine the role and functions of accessory sex glands and testes in Heliothis virescens in spermatophore formation and stimulation of egg maturation in mated females. Normal females mated to accessory sex glandectomized (-ASG) males produced fewer eggs and retained most of their eggs in ovarioles compared with females mated to sham-operated (+ASG) or normal males. The ASG are the source of the components for spermatophore formation in H. virescens. Females mated to castrated (-Testes) males showed similar pattern of egg production as did females mated to -ASG males. Females mated to -Testes males and those mated to sham-operated (+Testes) or normal males were found to have spermatophore in the bursa after uncoupling. Normal females mated to allatectomized (-CA) males developed similar numbers of total eggs as did females mated to sham-operated (+CA) males and normal males. PMID- 12770427 TI - Cuticular extracts inducing aggregation in the German cockroach, Blattella germanica (L.). AB - German cockroaches Blattella germanica (L.) are gregarious insects. An aggregation pheromone contributes to the maintenance of aggregates. Choice experiments checked the efficiency of different solvents, i.e. dichloromethane, methanol and pentane, in extracting attractive substances and compared the attractiveness of extracts of different parts of the body. Dichloromethane and pentane were the most efficient solvents tested for extracting the attractive substances. Methanol whole body extracts appeared inefficient to induce aggregation. The proportion of larvae attracted to conditioned papers decreased in relation to the size of cuticular surface washed, from whole body to half-body and again to a section of the body cut in three. Attractive substances appear to be present on all parts of the body. Chemical analysis by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) showed that the active extracts contained only cuticular hydrocarbons. In addition to behavioural tests, differences between the composition of methanol extracts and that of the extracts for the other two solvents were revealed by GC. These results indicated that the cuticular hydrocarbons operate as an aggregation pheromone in Blattella germanica. PMID- 12770428 TI - An improved aphid electroantennogram. AB - Excised antennae have been used to record aphid electroantennograms (EAGs) but these preparations have small, rapidly declining responses and a short usable life. An improved EAG technique is described and evaluated using alate virginoparae of the black bean aphid, Aphis fabae, and a series of plant volatiles. EAG recording with fine-tip glass electrodes inserted into the surface of the intact antenna of a whole insect preparation gave stable EAG peaks of a few mV and EAG peak amplitude decreased by only 22% over an 8 hour recording period. The EAG responses showed typical dose-dependent characteristics. There was variation in EAGs from different preparations, so normalisation of the EAG responses against a standard stimulus is still necessary. The thirty compounds tested elicited an EAG response profile largely similar to that reported previously from excised antennae. EAG responses recorded with the electrode at three different positions, between the 5th-6th (A), 4th-5th (B), and 3rd-4th (C) antennal segments, were smaller when recorded closer to the head. Position B produced larger EAG responses than those at C although there was no olfactory receptor between position B and C. PMID- 12770429 TI - Adaptation of hypopharyngeal gland development to the brood status of honeybee (Apis mellifera L.) colonies. AB - We studied pollen consumption, head weight, hypopharyngeal gland (HPG) acini diameter, and protein synthesis and transfer in honeybee workers reared in colonies with normal and with decreasing amounts of brood. We found that head fresh weight is correlated with size of the glands and that pollen consumption is positively correlated with gland development. An effect of brood on size of the glands could be confirmed, but was not as profound as in previous studies. Similarly, no difference in the amount of protein synthesized or transferred in workers living under the two brood conditions was found. We suspect this is due to the fact that HPGs also supply food to young bees and in our study young bees were always present while in previous studies, colonies often lacked both brood and young bees. PMID- 12770430 TI - Ecdysteroid release by the prothoracic gland of Gryllus bimaculatus (Ensifera: Gryllidae) during larval-adult development. AB - The in vitro secretion of ecdysteroids from the prothoracic glands of larvae of Gryllus bimaculatus was analysed by HPLC-RIA. The primary product was identified as 3-dehydroecdysone (65-93%), with lesser amounts of ecdysone (7-35%). Production and release of ecdysteroids from the prothoracic glands are calcium dependent. The rate of ecdysteroid release was low during the beginning and the end of the last two larval stages and high in between. Prothoracic glands from young adult females produced only minor amounts of ecdysteroids and ceased hormone production around day 4 after the moult. PMID- 12770431 TI - The role of surface characteristics in eliciting humoral encapsulation of foreign bodies in Plasmodium-refractory and -susceptible strains of Anopheles gambiae. AB - A refractory strain of the mosquito, Anopheles gambiae, melanotically encapsulates and kills many species of malaria parasites, whereas susceptible strains allow the parasites to develop normally. To study the role of surface characteristics in eliciting this immune response, 27 types of chromatography beads that differed in matrix type, charge, functional group, and functional group density were assayed for degree of melanotic encapsulation in refractory and susceptible mosquitoes. Overall, two glucan-based matrices, Sephadex (dextran) and cellulose, stimulated the strongest responses, regardless of functional group. Substituting matrix hydroxyl groups with functional groups on Sephadex and cellulose beads decreased the level of encapsulation. These results demonstrate that glucans induce melanotic encapsulation in An. gambiae. Beads with agarose, polystyrene, and acrylic matrices, and most methacrylate-based beads elicited little or no melanization; however, epoxide-methacrylate beads were encapsulated, demonstrating that glucans are not essential for eliciting a response. Comparisons between the two strains demonstrated that refractory mosquitoes melanized many bead types to a greater degree than did susceptible mosquitoes. On this basis, we propose that an important difference between the two strains is that one of the enzymes involved in the melanization pathway functions at a higher level in the refractory strain. Finally, of all beads tested, only 85% substituted CM-Sephadex beads were virtually unmelanized in susceptible mosquitoes but highly melanized in the refractory strain; thus, a specific surface microenvironment is necessary to demonstrate this effect. PMID- 12770432 TI - The effect of cold acclimation and deacclimation on cold tolerance, trehalose and free amino acid levels in Sitophilus granarius and Cryptolestes ferrugineus (Coleoptera). AB - Canadian and French laboratory strains of Sitophilus granarius (L.) and Cryptolestes ferrugineus (Stephens) were cold acclimated by placing adults at 15, 10 and 5 degrees C successively for 2wk at each temperature before deacclimating them for 1wk at 30 degrees C. Unacclimated S. granarius had an LT(50) (lethal time for 50% of the population) of 12days at 0 degrees C compared with 40days after the full cold acclimation. At -10 degrees C, unacclimated C. ferrugineus had an LT(50) of 1.4days compared with 24days after the full acclimation. Cold acclimation was lost within a week after returning insects to 30 degrees C. Trehalose, as well as the amino acids proline, asparagine, glutamic acid and lysine were higher in cold acclimated insects for both species. For S. granarius, glutamine was higher in cold acclimated insects and isoleucine, ethanolamine and phosphoethanolamine, a precursor of phospholipids, were lower in cold acclimated insects. For C. ferrugineus, alanine, aspartic acid, threonine, valine, isoleucine, leucine, phenylalanine and phosphoethanolamine were higher in cold acclimated insects. For both species tyrosine was lower in cold acclimated insects. There were small but significant differences between Canadian and French strains of S. granarius, with the Canadian strain being more cold hardy and having higher levels of trehalose. There were small but significant differences between male and female S. granarius, with males being more cold hardy and having higher levels of proline, asparagine and glutamic acid. In conclusion, high levels of trehalose and proline were correlated with cold tolerance, as seen in several other insects. However, correlation does not prove that these compounds are responsible for cold tolerance, and we outline further tests that could demonstrate a causal relationship between trehalose and proline and cold tolerance. PMID- 12770433 TI - Feeding effects on gene expression of the hypertrehalosemic hormone in the cockroach, Blaberus discoidalis. AB - Feeding effects on hypertrehalosemic hormone (HTH) transcript levels in corpora cardiaca (CC) of adult females of the cockroach, Blaberus discoidalis were measured using dot blot hybridization. HTH transcript levels were nearly doubled in CC from females withheld from food and water for ten days compared to CC from fed females. The increase in HTH-mRNA was a response to starvation, not dehydration, and reversed within 2 days after exposure to food. HTH-mRNA was elevated in CC from fed insects that had their recurrent nerve severed, but low fecal output by insects with severed nerves indicated that feeding and digestion were impaired. Thus, the increased HTH synthesis likely resulted from starvation rather than disruption of neural regulation. CC from starved females that were refed with either solutions or agar that contained glucose did not show down regulation of HTH-mRNA. Likewise, injections of glucose or trehalose did not suppress HTH-mRNA levels in CC of starving insects. Down-regulation of the starvation-related increase in HTH-mRNA appears to be a response to consumption of a complex of nutrients and not to increased carbohydrates or mechanical aspects of feeding. PMID- 12770434 TI - The action of inhibitors of protein kinases on fluid and ion secretion by Malpighian tubules of Locusta migratoria, L. AB - Fluid production in Locusta Malpighian tubules was stimulated by corpora cardiaca extract (c. 100%) and dibutyryl cAMP (c. 50%). Chelerythrine and staurosporine (Protein kinase C, PKC inhibitors) inhibited it in the range 0.07-60&mgr;M (IC(50)3&mgr;M), whereas Rp-cAMP (Protein kinase A, PKA inhibitor) caused inhibition over the concentration range 10-1000&mgr;M (IC(50)264&mgr;M). The protein phosphatase inhibitor, okadaic acid, was also inhibitory over the concentration range 0.1-1000nM (IC(50) 91nM). CC extract stimulation increased fluid [Na(+)] from 41 to 59mM and decreased [K(+)] from 127 to 107mM; stimulation with cAMP had no such effect. The PKC inhibitors reduced the [K(+)] in the secreted fluid from 126 to 107mM but had no effect on the [Na(+)]. Subsequent addition of CC extract stimulated fluid production and caused an increase in [Na(+)] from 41 to about 50mM. The addition of Rp-cAMP reduced fluid production but caused a decrease in [Na(+)] from 37 to 28mM and an increase in its [K(+)] from 124 to 148mM. Fluid production by Rp-cAMP inhibited tubules was not stimulated by corpora cardiaca extract or cAMP, but [Na(+)] rose to 36mM. Protein phosphorylation plays a role in the regulation of fluid production probably via the apical and basal membrane cation transporters. PMID- 12770435 TI - Degeneration of moulting glands in male crickets. AB - The degeneration of the prothoracic glands of the male cricket, Gryllus bimaculatus, was analyzed by using an in vitro assay for ecdysteroid release from the moulting glands in last instar nymphs as well as in adult animals, and correlated with light and transmission electron microscopy. Apoptosis was examined by the TUNEL-reaction. The ability to synthesize ecdysteroids reached a peak at the 8th day of the last larval instar, identified as the moulting peak. After adult ecdysis it decreased to barely measurable values. Prothoracic gland degeneration was initiated at the time of the moulting peak, characterized by TUNEL positive reactions, nuclear and cytoplasmatic condensation, a striking abundance of residual basal laminae; besides a great amount of autophagic vacuoles are observed. The results reveal that apoptosis and autophagy are the basic mechanisms for programmed cell death in the prothoracic gland of Gryllus bimaculatus. PMID- 12770436 TI - Temperature dependence and acclimatory response of amylase in the High Arctic springtail Onychiurus arcticus (Tullberg) compared with the temperate species Protaphorura armata (Tullberg). AB - Amylolytic activity was measured in whole body homogenates of High Arctic (Onychiurus arcticus) and temperate (Protaphorura armata) springtails (Collembola: Onychiuridae) in the temperature range 5-55 degrees C. A pH of ca. 8 was optimum for amylolytic activity in both species. A higher weight-specific amylolytic activity was observed in P. armata. In O. arcticus, amylolytic activity depended on thermal acclimation, which increased during 2 and 9 weeks of cold acclimation (5 degrees C) and decreased over 7 weeks of warming (15 degrees C) of animals that were previously acclimated to cold for 2 weeks. In cold acclimated O. arcticus, a slower decrease of amylolytic activity occurred with lowering of temperature in the range 5-20 degrees C in comparison with warm acclimated specimens and P. armata, which resulted in higher activity at 5 degrees C. The activation energy calculated from an Arrhenius plot for P. armata was 68.7 kJ.mol(-1). In O. arcticus it was between 30.2 and 61.5 kJ.mol(-1), being lower in cold-acclimated samples. The temperature optimum for amylolytic activity was higher in the temperate species (40 degrees C), whilst in O. arcticus it depended on the acclimation regime: it rose to 35 degrees C after warm acclimation and decreased to 20 degrees C after cold adaptation. The total soluble protein content of body tissues of O. arcticus also increased during cold acclimation. These differences between the two species suggest that amylolytic activity is an indicator of cold adaptation in the High Arctic O. arcticus. PMID- 12770437 TI - The chemical composition of the spermatophore in some species of phaneropterid bushcrickets (Orthoptera: Tettigonioidea). AB - Male bushcrickets transfer large spermatophores of up to 40% of male body mass to the females during mating. These large nuptial gifts are later consumed by the female and have been shown to affect the size and number of eggs laid after mating in some species. The composition of the spermatophores in five species of phytophagous phaneropterid bushcrickets (genera Ancistrura, Barbitistes, Metaplastes, Poecilimon) was examined in respect to water content, elemental composition, protein concentration, and the content of lipids, carbohydrates, urea and uric acid. In addition, the amino acid composition of the spermatophore was compared with that of the egg proteins. In all species, water content was found to be about 85% of wet mass. Eleven to 16% of the dry mass consisted of nitrogen, corresponding to a protein content of about 70 to more than 90% of the dry mass. Urea and uric acid were only present in traces. The proteins contained a high amount of glycine (about 26mol %), together with asparagine/aspartic acid, 12% and glutamic acid, 11%, which differed distinctly from the amino acid composition of the egg proteins. The results are discussed with respect to the advantages the male may receive by producing these large nuptial gifts. PMID- 12770438 TI - Accumulation of Hsp70 mRNA under environmental stresses in diapausing and nondiapausing adults of Drosophila triauraria. AB - Drosophila triauraria entered reproductive diapause in response to short daylengths and acquired tolerance to heat, cold and desiccation. In this species, the heat-shock response (accumulation of Hsp70 mRNA in response to heat) occurred at 27-41 degrees C, and the level of Hsp70 mRNA did not differ between diapausing and nondiapausing individuals. Hsp70 mRNA was also induced by exposure to -4 or 8 degrees C. However, it was scarcely detected just after the exposure to cold, but accumulated when flies were maintained at normal temperature following the exposure to cold. The level of Hsp70 mRNA was lower in diapausing individuals than in nondiapausing ones when exposed to -4 degrees C, but was not different between them when exposed to -8 degrees C. This species did not synthesize Hsp70 mRNA under desiccation stress irrespective of the diapause state. These results suggest that diapausing individuals of this species acquired tolerance to heat, cold and desiccation independent of the transcriptional regulation of the hsp70 gene PMID- 12770439 TI - Caterpillars have evolved lungs for hemocyte gas exchange. AB - Since insect blood usually lacks oxygen-carrying pigments it has always been assumed that respiratory needs are met by diffusion in the gas-filled lumen of their tracheal systems. Outside air enters the tracheal system through segmentally arranged spiracles, diffuses along tubes of cuticle secreted by tracheal epithelia and then to tissues through tracheoles, thin walled cuticle tubes that penetrate between cells. The only recognized exceptions have been blood cells (hemocytes), which are not tracheated because they float in the hemolymph. In caterpillars, anoxia has an effect on the structure of the hemocytes and causes them to be released from tissues and to accumulate on thin walled tracheal tufts near the 8th (last) pair of abdominal spiracles. Residence in the tufts restores normal structure. Hemocytes also adhere to thin-walled tracheae in the tokus compartment at the tip of the abdomen. The specialized tracheal system of the 8th segment and tokus may therefore be a lung for hemocytes, a novel concept in insect physiology. Thus, although as a rule insect tracheae go to tissues, this work shows that hemocytes go to tracheae. PMID- 12770440 TI - Factors affecting the freeze tolerance of the hoverfly Syrphus ribesii (Diptera: syrphidae). AB - Larvae of Syrphus ribesii collected from overwintering sites in the U.K. are strongly freeze tolerant with 70% survival at -35 degrees C. The cold tolerance of laboratory reared insects increased with increasing periods of acclimation at 0 degrees C, with a concurrent rise in the supercooling point (SCP) from -6.8+/ 0.1 to -5.1+/-0.3 degrees C. There was 50% survival in the most cold-hardy group 72h after brief exposures to -30 degrees C. The retention of gut contents caused a decrease in cold hardiness, with only 13% of larvae surviving 72h after exposure to -15 degrees C, with no subsequent pupation or emergence. Wet larvae had a significantly higher SCP (-5.0+/-0.2 degrees C) compared to dry larvae ( 7.8+/-0.4 degrees C), although survival of larvae was similar in both groups. There was no nucleator activity in the haemolymph of field collected larvae. The importance of these findings are discussed in relation to the freeze tolerance strategy of S. ribesii. PMID- 12770441 TI - Feeding and activation of corpora allata in the cockroach Blattella germanica (L.) (Dictyoptera, Blattellidae). AB - Adult females of the cockroach Blattella germanica have clearly-defined feeding cycles related to oogenesis. In the first cycle, food ingestion precedes volumetric increase in the corpora allata, which in turn precedes juvenile hormone production, whereas starved females do not develop the corpora allata and produce very low amounts of juvenile hormone. When the second gonadotropic cycle is provoked by removing the ootheca, the first event observed is an increase in food consumption, followed by an increase in corpora allata volume and activity. However, this increase in corpora allata volume (and activity) does not occur if females are starved, thus indicating that the ootheca in the genital chamber inhibits primarily feeding, and indirectly corpora allata development and activity. Corpora allata volume in isolated heads from starved and decapitated females was able to increase to levels similar to fed controls, but this increase was abolished by allatostatin treatment. We suggest that a factor produced in the thoracico-abdominal compartment, which reaches the head mainly through a nervous pathway, is released during starvation and inhibits corpora allata development. This factor may stimulate allatostatin production or release, or may well be allatostatin itself. PMID- 12770442 TI - Electrochemical characteristics of ion secretion in malpighian tubules of the New Zealand alpine weta (Hemideina maori). AB - Characteristics of ion and fluid secretion were investigated in isolated Malpighian tubules of the New Zealand Alpine Weta (Hemideina maori). Fluid secretion by tubules in iso-osmotic saline (500mOsm) occurred at a rate of 15+/ 3nlh(-1) and was enriched in K(+) (approx. 125mmoll(-1)) relative to the saline (10mmoll(-1)). Maximal fluid secretion (112nlh(-1)) during simultaneous exposure to hypo-osmolality and dibutyryl cAMP resulted in an 8.8x increase in the quantity of K(+) secreted, compared to only a 2.4x increase in Na(+) secretion. Measurements of intracellular ion activities and membrane potentials indicated that Na(+) and K(+) were transported against a strong electrochemical gradient across the apical surface, regardless of saline osmolality. On the basolateral surface, there was a large driving force for Na(+) entry, while K(+) was distributed near its equilibrium potential. Neither bumetanide nor ouabain in the bathing saline had a significant effect on fluid secretion, but Ba(2+) and amiloride decreased fluid secretion by 79 and 57%, respectively. The effect of Ba(2+) on fluid secretion was consistent with a high basolateral permeability to K(+), relative to Na(+) and Cl(-). These results indicate that the characteristics of fluid secretion in this primitive insect are largely conserved with characteristics reported for other insects. PMID- 12770443 TI - Detection of major and minor sex pheromone components by the male codling moth Cydia pomonella (Lepidoptera: Tortricidae). AB - Electroantennograms (EAGs) recorded from the antennae of male Cydia pomonella L. in response to stimulation with doses of the main sex pheromone component E8,E10 dodecadienol (Codlemone) ranging from 5ng to 500&mgr;g did not differ in their amplitudes from responses obtained to a synthetic 7-component pheromone blend containing the same absolute quantities of Codlemone. Based on differences in spike amplitudes obtained in Single Cell recordings (SCR), Sensilla trichodea on the antenna of males were found to contain at least three receptor neurone types. Two olfactory receptor neurones were tuned to Codlemone, while the third failed to be stimulated by Codlemone or by the minor components of the pheromone blend. As spike activity of the neurones in the S. trichodea stimulated by the 7 component blend did not differ from that of stimulation by Codlemone alone it appears that none of the receptor neurones is sensitive to any of the minor components tested. Scanning-electron-microscopical (SEM) examination of Sensilla auricillica on the antennae of Cydia males revealed two morphologically distinct types: rabbit eared shoehorn and regular shoehorn. SCR from these sensilla showed that only olfactory receptor neurones located in the rabbit-eared shoehorn type were tuned to the minor components. Differences in spike amplitudes (large, intermediate, small) allowed three types of neurones to be distinguished. Only the spike frequency of the intermediate receptor neurone was increased by application of the minor components E8-dodecenol, E9-dodecenol, dodecanol, tetradecanol, hexadecanol and E8,E10-dodecadienal. None were stimulated by Codlemone. These results are discussed in relation to the behavioural role of the minor pheromone components of C. pomonella. PMID- 12770444 TI - Differential adsorption of allospecific hydrocarbons by the cuticles of two termite species, Reticulitermes santonensis and R. lucifugus grassei, living in a mixed colony. Passive transfer by contact. AB - When members of the two termite species Reticulitermes santonensis and Reticulitermes lucifugus grassei were placed together, each species acquired some of the allospecific cuticular products. When living individuals of each of these two species were placed together, their cuticular hydrocarbon profiles changed very quickly, since it was within the first two hours of cohabitation that they differed most from those of the corresponding control individuals. After the first two hours, the profiles of the R. santonensis individuals continued to change only very little if at all, whereas 24h later, the process of change continued in the R. lucifugus grassei individuals until their profiles resembled those of the mixed R. santonensis individuals more than their own original profiles. The profiles of the R. l. grassei individuals therefore underwent a greater change than those of the R. santonensis individuals during the period of cohabitation. The fact that similar results were obtained when dead members of these two species were placed together suggests that this difference in the adsorption of allospecific hydrocarbon by the cuticles of the members of the two species cannot be attributable to any behavioural differences, but to differences in physico-chemical composition of the cuticles between the two species. PMID- 12770445 TI - Developmental changes of storage proteins and biliverdin-binding proteins in the haemolymph and fat body of the common cutworm, Spodoptera litura. AB - The concentrations of three storage proteins (SL-1,SL-2 and SL-3, hexamers of 70 80kDa subunits) and two biliverdin-binding proteins (BP-A and BP-B, dimers of 165kDa) in the haemolymph and fat body during larval and pupal development of Spodoptera litura were determined by immunodiffusion tests using polyclonal antisera. SL-1 and SL-2 (methionine-rich) first appeared in the haemolymph of one day-old sixth (final) instar larvae, prominently increased in the haemolymph during the later feeding period and were almost totally sequestered by the fat body after gut purge. SL-3 (arylphorin) was first detected in the haemolymph during the molting period to the final larval ecdysis, increased in concentration throughout the entire feeding period of the final larval instar and was partly sequestered by the fat body several hours later than the other storage proteins. BP-A showed nearly the same pattern in the haemolymph as SL-3: BP-B increased during feeding period and decreased during molting period and attained a maximum level during the penultimate larval instar, however its concentration decreased considerably and remained low in the final larval instar. BP-A was partly and BP B was almost totally sequestered by the fat body 8 h after sequestration of SL-1 and SL-2, rendering the fat body blue in colour. These facts suggest an additional function of biliverdin-binding proteins as amino acid storage proteins and the results show a differential uptake mechanism for these proteins by the fat body. PMID- 12770446 TI - The functional significance of switching interval in food mixing by Locusta migratoria. AB - To investigate the consequences of different patterns of switching between nutritionally complementary foods, nymphs of Locusta migratoria L. (Orthoptera: Acrididae) were forced to alternate between pairs of artificial foods differing only in their protein and carbohydrate content at one of several fixed intervals. Total food and nutrient intake over 6days were very similar in all groups despite the considerable variation in temporal patterns of nutrient intake. The effects of these treatments were determined using measurements of growth, stadium duration and carcass composition. Those given a low variance food pairing showed little effect in any of the measures while those given a high variance pairing showed no differences in dry weight growth or stadium duration but did differ in protein and lipid content. There was little evidence from our measurements that an ad-libitum alternation time of 4h previously observed in a laboratory experiment was physiologically optimal, suggesting that ecological factors may underlie such behaviour. Possible compensatory mechanisms used by L. migratoria are suggested. PMID- 12770447 TI - Factors affecting carbohydrate and free amino acid content in overwintering larvae of Enosima leucotaeniella. AB - Amounts of several metabolites were measured in overwintering larvae of Enosima leucotaeniella acclimated to temperatures between -5 and 15 degrees C for 30days. In the diapausing stage, cold hardiness, as shown by the survival rate, began rising below 15 degrees C. Glycogen content decreased as the temperature decreased from 10 to 0 degrees C. Trehalose content rose as the temperature decreased from 15 to 5 degrees C, but remained unchanged as the temperature decreased from 5 and 0 degrees C. Twenty-eight free amino acids were detected in the haemolymph; levels of proline, glutamine and glutamic acid increased at high temperatures, but alanine increased at low temperatures, especially as temperature decreased from 5 to 0 degrees C. Lipid content was unchanged by the different acclimation temperatures. The effects of temperature, diapause and aerobic conditions on the levels of carbohydrates and amino acids in overwintering larvae were analyzed. Alanine levels rose at low temperature only when the larvae were in the diapausing stage. The level of trehalose rose at low temperature in both the diapausing and post-diapausing stages, although it was higher at aerobic conditions in the post-diapausing stage. These results suggest that efficient trehalose synthesis occurs under the combination of low temperature and aerobic conditions of the post-diapausing stage, so that cold hardiness in overwintering E. leucotaeniella larvae may rise to a high level in winter. PMID- 12770448 TI - Summer and winter diapause in pupae of the cabbage butterfly, Pieris melete Menetries. AB - The cabbage butterfly, Pieris melete is multivoltine with a pupal summer and winter diapause. Summer and winter diapause are induced principally by relatively long and short daylengths, respectively. The intermediate to relatively short daylengths of autumn permitted some pupae to develop without diapause in the field. A short daylength had a stronger diapause inducing effect than a relatively long one under higher temperatures. The principal sensitive phase for photoperiodic response occurred before the late 3rd larval instar. The critical daylength for wild autumnal populations was between 12h 30min and 12h 40min at an average temperature of 20.5 degrees C. A night interruption by 2h of light averted diapause most effectively when it was placed 10 to 12h after lights-off. High temperatures and long days during summer inhibited the incidence of diapause, suggesting that the occurrence of summer diapause is due to the specific climatic conditions occurring in April and early May, rather than to the high temperatures in summer. This indicates that the butterfly has a cryptic ability to reproduce in summer. High temperatures delayed diapause development, whereas low temperatures enhanced it, indicating that the optimum temperature of diapause development is lower. The diapause regulating mechanisms thus ensure that the species synchronises its development and reproduction with the growth seasons of the host plants and provide the species with a high degree of flexibility in its life cycle. PMID- 12770449 TI - Inheritance of the photoperiodic response controlling larval diapause in the blow fly, Calliphora vicina. AB - Larvae of the blow fly Calliphora vicina R-D. (Diptera: Calliphoridae) display a diapause in response to the exposure of their parents to short photoperiods. Due to geographic variation in photoperiodic response, flies from a southern, English population show a long-day response to the fixed photoperiod of L:D 15.5:8.5 whilst flies from a northern population from Finland show a short-day response to the same photoperiod. Crosses between these strains have shown previously that diapause incidence is a maternal characteristic; here we demonstrate that the hybrid female offspring of such crosses are not intermediate between the two parental strains but show a photoperiodic response biased towards their maternal line. Thus not only are males unable to influence directly the diapause incidence among their offspring but the indirect effects of inheritance down the male line are weaker than down the female. Diapause duration, in contrast, is influenced by each parent in a similar manner. Diapause lasts longer in larvae with a greater admixture of northern genes regardless of whether they were maternal or paternal. PMID- 12770450 TI - Hemolymph juvenile hormone titers in pupal and adult stages of southwestern corn borer [Diatraea grandiosella (pyralidae)] and relationship with egg development. AB - Juvenile hormones I, II and III were monitored in hemolymph of pupal and adult stages of various ages of Diatraea grandiosella females. JH III was the predominant homologue followed by JH II, and JH I was rarely detectable. At day 5 after pupation, no JH was detectable. JH titers increased from 7.5days after pupation to a peak of 24.8ngml(-1) JH II and 26ngml(-1) JH III at adult emergence and then declined to low levels by 24h after emergence. Ovarian development in D. grandiosella parallels changes in hemolymph JH titers, but the role of JH in vitellogenesis is unclear since the time of vitellogenesis initiation has yet to be determined. No apparent vitellogenin deposition was observed in eggs 5days after pupation. Some oocytes were partially vitellogenic by 7.5days after pupation and oocytes continued to grow afterwards, but no oocytes were chorionated during the pupal stage. Chorionated oocytes were observed in 24-h-old female moths. Juvenile hormone is essential for chorion formation in this species, because decapitated pupae treated with 10&mgr;g JH III in corn oil developed chorionated oocytes while decapitated pupae treated with corn oil did not. PMID- 12770451 TI - Effects of snowdrop lectin (GNA) delivered via artificial diet and transgenic plants on the development of tomato moth (Lacanobia oleracea) larvae in laboratory and glasshouse trials. AB - The effects of snowdrop lectin (Galanthus nivalis agglutinin, GNA) on Lacanobia oleracea larval growth, development, consumption, and survival, were examined by 3 distinct bioassay methods. Larvae were reared on artificial diet containing GNA at 2% (w/w) dietary protein; on excised leaves of transgenic potato expressing GNA at approx. 0.07% of total soluble proteins; and on transgenic potato plants expressing GNA at approx. 0.6% of total soluble proteins in glasshouse trials. Significant effects on larval growth were observed with all three treatments. At 21days after hatch mean larval biomass was reduced by 32 and 23%, in the artificial diet and excised leaf bioassays respectively. In glasshouse trials a 48% reduction in insect biomass per plant was observed after 35days. The artificial diet and excised leaf assays also showed that GNA significantly slowed larval development as assessed by instar duration. GNA caused a 59% overall reduction in mean daily consumption in the artificial diet assay, and a significant reduction in leaf damage in glasshouse trials. However, prolonged compensatory feeding by larvae in the excised leaf assay resulted in their consuming 15% more total leaf material than the control group. Adaptation to low levels of GNA, in terms of biomass recovery and compensatory feeding, was observed within one larval generation in the detached leaf assay. No significant effects of GNA on larval survival were observed in the artificial diet and detached leaf bioassays, whereas survival was decreased by approx. 40% in the glasshouse bioassay. The assays show that the insecticidal effects of GNA can be observed both in vitro when fed in artificial diet and in planta, and can be demonstrated in the glasshouse as well as under growth cabinet conditions. PMID- 12770452 TI - Vision of the honeybee Apis mellifera for patterns with one pair of equal orthogonal bars. AB - The visual discrimination of patterns of two equal orthogonal black bars by honeybees has been studied in a Y-choice apparatus with the patterns presented vertically at a fixed range. Previous work shows that bees can discriminate the locations of one, or possibly more, contrasts in targets that are in the same position throughout the training. Therefore, in critical experiments, the locations of areas of black were regularly shuffled to make them useless as cues. The bees discriminate consistent radial and tangential cues irrespective of their location on the target during learning and testing. Orientation cues, to be discriminated, must be presented on corresponding sides of the two targets. When orientation, radial and tangential cues are omitted or made useless by alternating them, discrimination is impossible, although the patterns may look quite different to us. The shape or the layout of local cues is not re-assembled from the locations of the bars, even when there are only two bars in the pattern, as if the bees cannot locate the individual bars within the large spatial fields of their global filters. PMID- 12770453 TI - On the respiratory quotient (RQ) of termites (Insecta: Isoptera). AB - The respiratory quotient (RQ) at 28 degrees C was determined by Warburg manometry in 23 species of termites from the Mbalmayo Forest Reserve (Cameroon) and three sub-tropical species cultured under laboratory conditions in the U.K. or freshly collected in Australia. The data are tabulated with other recently reported RQs (determined by manometry or GC) and with measured CH(4) emission rates to provide a survey of 29 species covering both lower and higher termites in all major trophic (functional) categories. In all species, except the wood-feeding Coptotermes acinaciformis and the soil-feeding Cubitermes fungifaber, the observed mean values (with manometry corrected for known fluxes of H(2) and CH(4)) were at or well above 1.00. Soil-feeding forms (except C. fungifaber) generally showed a high apparent RQ (not corrected for H(2)), with nine species (out of 13) above 1.20 and six species above 1.30. Well-replicated laboratory experiments with Reticulitermes lucifugus showed that there was a tendency for RQ to fall with time over a 4-h incubation, although remaining greater than 1.00.The observed RQs are consistent with carbohydrate being the principal substrate supporting respiration in all trophic and taxonomic categories, with little or no contribution from the degradation of lignin or other polyaromatic materials. However, in many species (especially soil-feeders), the observed RQ is greater than that expected from known fluxes of O(2), CO(2) and CH(4) on the assumption that carbohydrate is the respiratory substrate. This presupposes that there is a large hydrogen sink (additional to CH(4) production), possibly the emission of H(2) gas, and/or the existence of unresolved digestive mechanisms or electron routings. Uncertainties in the use of manometry with termites are discussed. PMID- 12770454 TI - Rapid cold hardening in the western flower thrips Frankliniella occidentalis. AB - A rapid cold hardening process is reported in first instar larvae of Frankliniella occidentalis. When larvae are transferred directly from 20 degrees C to -11.5 degrees C for 2h there is 78% mortality, whereas exposure to 0 degrees C for 4h prior to transfer to -11.5 degrees C reduces mortality to 10%. The response can also be induced by exposure to 5 degrees C for 4h or by gradual cooling at rates between 0.1 and 0.5 degrees C min(-1.) The acquired cold tolerance is transient and is rapidly lost (after 1h at 20 degrees C). Rapid cold hardening extends survival times at -11.5 degrees C and depresses lethal temperatures in short (2h) exposures. Rearing at 15 degrees C (12L:12D), (a cold acclimation regime for F. occidentalis), does not protect against the cold shock induced by direct transfer to -11.5 degrees C (which rapid cold hardening does) but does extend survival time at -5 degrees C (i.e. increased chill tolerance) whilst rapid cold hardening does not. The rapid and longer term cold hardening responses in F. occidentalis therefore appear to have different underlying mechanisms. PMID- 12770455 TI - Photoperiodism and control of summer diapause in the Mediterranean tiger moth Cymbalophora pudica. AB - Photoperiodic responses to both constant and changing photoperiods were studied in the Mediterranean tiger moth Cymbalophora pudica. Embryos, larval instars and prepupae were all sensitive to photoperiod, and the responses of larvae and prepupae to changing photoperiods were similar. At 23+/-2 degrees C, constant 24 h photoperiods with short photophases (11, 12h) induced a long diapause (mean 88days) whereas long photophases (14, 16h) induced a short diapause (mean 52days). A change to a longer or shorter photophase during larval development or during diapause caused a significant extension (up to a maximum of 138days) or shortening (down to a minimum of 10days) of diapause, respectively, but only when at least one of the photophases was longer than 14h. Thus, shortening and prolongation of photophase had an opposite effect than constant short and long photophases, respectively. Changes within the range of photophases of 10-14h did not cause a significant change in diapause duration.Experimental results enabled us to outline the mechanisms regulating voltinism and the duration of summer diapause. For the monovoltine cycle, cold autumn/winter temperatures slow down larval development, and prepupal aestivation starts relatively late (March, April). Prepupae are then kept in diapause by the increasing daylength (>14h after late April). Pupation is synchronized by decreasing daylength after summer solstice, and imagoes emerge in September/October. For the bivoltine cycle, when the autumn/winter temperatures are relatively warm, a certain portion of the population (depending on the individual rate of growth) may be diverted to a bivoltine life-cycle. In such a case, larval development is fast and short enough to allow an early start of diapause (prior to or during February). The duration of such early diapause is not influenced by changes in daylength (<14h); pupation occurs very early (April/May), and spring generation imagoes fly and oviposit in May/June. Summer larvae and prepupae live under decreasing daylength (>14h), which shortens their diapause to 20-30days. Imagoes of the autumnal generation thus occur in September/October, together with the univoltine portion of the population. PMID- 12770456 TI - An analysis of resource allocation in response to dietary yeast in Drosophila melanogaster. AB - Drosophila melanogaster exhibit an increase in fecundity and a decrease in length of life and starvation resistance when the diet is enriched through the addition of live yeast. It has been proposed that this represents a necessary energetic trade-off between reproductive and somatic functions. We examined the metabolic aspects of this trade-off. We measured egg production, dry wt, somatic lipid and carbohydrate storage, and metabolic rate in response to changing amounts of live dietary yeast. These variables were measured in five replicate populations selected for postponed aging and five replicate short lived control populations. We find that all ten populations show an overall increase in egg production and decrease in the amount of stored metabolites in the presence of increasing amounts of yeast. All populations increase metabolic rate in response to increasing amounts of live dietary yeast. The energetics of this phenomenon suggest that increased egg production results from increased acquisition of nutrients available in yeast with only a small redirection of resources from storage to egg production. PMID- 12770457 TI - Cycling of ecdysteroid levels in adult female stable flies, Stomoxys calcitrans in relation to blood feeding. AB - The effect of blood-feeding on total and specific immunoreactive ecdysteroids in Stomoxys calcitrans adult females was examined following the fourth and fifth blood meals when total whole body and hemolymph ecdysteroids showed a dramatic increase in the titer. In general, for both total and specific immunoreactive ecdysteroids that included highly polar material, 20,26-dihydroxyecdysone, 20 hydroxyecdysone and ecdysone, there were clear differences between the effects of the fourth and fifth meals. Following the fifth meal, the titers rose sooner, reached higher levels and remained high longer than those following the fourth meal. This is the first examination of the effects of back-to-back blood meals on total and specific ecdysteroid levels in an intermittent, blood-feeding fly. These results suggest that both rates of synthesis and degradation are affected by blood-feeding and that the number and possibly quantity of blood ingested affect the biochemical mechanisms that regulate ecdysteroid titers in S. calcitrans. PMID- 12770458 TI - The adaptation of insects to plant protease inhibitors. AB - Plants and herbivores have been co-evolving for thousands of years, and as a result, plants have defence mechanisms that offer protection against many herbivores such as nematodes, insects, birds and mammals. Only when a herbivore has managed to adapt to these defence mechanisms does it have the potential to become a pest. One such method of plant defence involves the production of protease inhibitors (PIs). These inhibitors are proteins that may be found constitutively in various parts of the plant, or may be induced in response to herbivore attack. PIs work at the gut level, by inhibiting the digestion of plant protein. This review focuses on insect herbivores and looks at the mechanisms involved in the role and function of PIs in plant defense against insects, as well as at the ability of well adapted species to overcome the effects of these plant PIs. PMID- 12770459 TI - Further evidence that diapause in the gypsy moth, Lymantria dispar, is regulated by ecdysteroids: a comparison of diapause and nondiapause strains. AB - A nondiapause strain of the gypsy moth offers an additional tool for evaluating the regulation of diapause in this species. Patterns of protein expression in the gut and gut enzyme activity distinguished the two strains. Synthesis of a 55kDa gut protein, previously linked to diapause, began 14days after oviposition in both the diapause (D) and nondiapause (ND) strains. Though synthesis of this protein persisted in the D strain, its synthesis decreased after day 18 in the ND strain. In the D strain, activity of the proteolytic enzymes (trypsin, chymotrypsin, elastase, aminopeptidase) and esterase remained low, while activity of all of these enzymes increased dramatically in the ND strain 18-20days after oviposition. By contrast, alkaline phosphatase (ALP) activity was high in both strains 15-17days after oviposition, activity remained high in the D strain but in the ND strain activity then decreased. Patterns of ALP zymograms were similar in the two strains on day 15, but later a band of high mobility appeared only in the D strain. When 20-hydroxyecdysone was added to hanging drop cultures containing ND pharate larvae 15days after oviposition, the larvae assumed the characteristics of diapause larvae: the 55kDa gut protein was synthesized, the ALP zymogram revealed the characteristic diapause pattern, and they failed to ingest culture medium. The fact that 20-hydroxyecdysone could elicit these responses in ND individuals further supports previous results indicating that ecdysteroids promote the induction and maintenance of the pharate larval diapause in this species. PMID- 12770460 TI - Responses to desiccation in four Coleopterans from sub-Antarctic South Georgia. AB - RATES OF WATER LOSS WERE DETERMINED FOR FOUR COLEOPTERA SPECIES: the herbivores Hydromedion sparsutum, Perimylops antarcticus (Family Perimylopidae) and the carnivores Trechisibus antarcticus, Oopterus soledadinus (Family Carabidae) collected during summer from a range of terrestrial habitats at South Georgia. A recording microbalance enabled measurement of individual weight loss with time in <5% r.h. at 10, 20, 30 and 35 degrees C. Adults of T. antarcticus had significantly higher rates of water loss than any other species over all temperatures. Individuals of both herbivores exhibited the slowest water loss rates under the experimental conditions. Within species, rates at 10 or 20 degrees C were slower than at the higher temperatures. Adult P. antarcticus had significantly greater amounts of body water than adult H. sparsutum for each of the four temperatures. Within species and life-stages of both herbivores, body water contents after drying at 10 degrees C were significantly lower than individuals dried at 30-35 degrees C, but no such differences were observed for the carnivores. At each temperature, rates of water loss were negatively correlated with initial live weight in all four species, but this was not the case within species or between adults and larvae. Maximum survival times during desiccation declined as temperature increased, but did not differ between species at 10 degrees C. Over 30-35 degrees C, survival times of both herbivores were significantly longer than either of the carnivores. Smaller insects (e.g. the carabids) had faster rates of water loss than the larger perimylopids under the same environmental conditions. The latter had greater resistance to desiccation than the former. It is suggested that the larger body water content of P. antarcticus enables it to resist desiccation more than the other three species, which correlates with its ecological distribution. Differences in water contents after drying individuals at low and high temperatures may be caused either by the water binding properties of cells and tissues or by reduction in energy stores in order to maintain metabolism at lower environmental temperatures causing a body weight loss. Whilst both herbivores show some physiological adaptations to drying conditions, it is suggested that the two carnivorous beetles may have adapted behaviourally to the South Georgia environment. PMID- 12770461 TI - Developmental changes in teratocytes of the braconid wasp Cotesia congregata in larvae of the tobacco hornworm, Manduca sexta. AB - The endoparasitic wasp Cotesia congregata develops in the hemocoel of larval stages of the tobacco hornworm, Manduca sexta. Teratocytes were released from the serosal membrane during hatching of the first instar wasp larva at 2-3days after oviposition; about 160 cells were released per embryo. The cells increased in diameter from about 10 to >200&mgr;m prior to wasp emergence. Nascent microvilli, visible on the cell surface before hatching of the first instar larva, rapidly increased in length and number following release of the cells. Irrespective of when the wasps were due to emerge, or how many parasitoids were present in the host, dramatic cytological changes occurred in the cells during the last instar of the host's development. Many of these morphological and ultrastructural changes were symptomatic of the cytological features of degenerating or apoptotic cells, and large numbers of vesicles appeared interspersed amongst the microvilli. The nucleus developed extensive dentritic ramifications, and the chromatin condensed in large clumps on the inner nuclear membrane. At the final stages of the wasps' development, the nucleus occupied the bulk of the interior of the cell. The cytoplasm gradually grew dramatically more electronluscent and less granular, as did the nucleoplasm, which is also indicative of impending cell death. Following the parasites' emergence, many of the cells underwent extensive blebbing of the cell surface. Teratocytes within a host appeared heterogeneous with respect to their morphological appearance. Analysis of the proteins secreted by teratocytes in vitro following labelling with (35)S-methionine showed that many (>30) polypeptides were synthesized de novo and secreted by the cells; some proteins were clearly targeted for secretion. We presume that the cells likely secrete a large number of proteins in vivo as well as in vitro. PMID- 12770462 TI - Comparison of the myotropic activity of position-2 modified analogues of proctolin on the hindgut of Periplaneta americana and the oviduct of Locusta migratoria. AB - The largest series of position-2 modified proctolin analogues to have been examined to date were tested for their ability to mimic the basal contraction induced by proctolin on hindgut of the cockroach, Periplaneta americana, and oviduct of the locust, Locusta migratoria. Twelve analogues of proctolin (Arg-Tyr Leu-Pro-Thr), differing in the substituent (H, OMe, OEt, OPr, F, Cl, Br, I, NO(2), NH(2), N(3), Me) located at the para-position of the aromatic amino acid, caused dose-dependent contractions of both tissues at concentrations quite similar to proctolin. Seven showed greater or equal potency on the hindgut but, with one exception, they were less active on the oviduct than proctolin. The rank order of potency of the analogues depends on the tissue, lending more support to the notion that insects have more than one type of proctolin receptor. No relationship was observed between myoactivity and lipophilic, steric, electron donating or electron withdrawing properties of the substituents at the para position of the aromatic amino acid. This may be the result of more than one sub type of proctolin receptor on the specific tissue with differing structural requirements for optimum activity. PMID- 12770463 TI - Effect of host nutritive value on egg production by Ceratitis capitata (Diptera, Tephritidae). AB - In a previous study we demonstrated that Ceratitis capitata larvae show a superior performance when they feed on the apical portion of papaya compared to larvae that feed on the basal portion, probably due to the higher levels of sugars in the apical portion. In the present study we carried out experiments to determine if larval feeding on the basal or apical portion of the fruit influences ovarian development and egg production by adult C. capitata females. The results indicate that females that feed on the apical portion during the larval stage have more developed ovaries and produce more eggs than females who feed on the basal portion during the larval stage. We also compared artificial diets of quantitatively similar chemical composition to the apical and basal portions of the papaya fruit. In this last experiment, no differences were detected in any of the parameters studied, i.e. percent emergence, time to pupation, adult size, larval preference, ovary size, egg production and estimated ingestion. PMID- 12770464 TI - Substrate vibrations elicit defensive behaviour in leafminer pupae. AB - Late instar larvae and pupae of the spotted tentiform leafminer Phyllonorycter malella (Ger.) (Lepidoptera: Gracillariidae) react with defensive behaviour when attacked by one of their parasitoids, the eulophid wasp Sympiesis sericeicornis Nees (Hymenoptera: Eulophidae). Vibrations produced during the insertion of the ovipositor into the mine are known to be important cues by which larvae detect the presence of their enemies. The aim of this study was to investigate which frequency components elicit defensive reactions in leafminer pupae using synthetic vibrations. Sine vibrations and bandlimited noise stimuli were offered to both free pupae and pupae concealed in their leafmines. Using Laser vibrometry we measured the vibrations experienced by pupae inside their mines and assessed the influence of the mine. Pupae were shown to react to substrate vibrations, and do so over a broad range of frequencies. Behavioural reactions to noise stimuli were stronger than to pure sine stimuli. Mine tissue attenuated vibration amplitudes of the input signal from 5.1 to 22.6dB. However, as response thresholds of concealed pupae were only twice as high as thresholds of free pupae (which is adequate to 3dB) pupae inside their mine were more sensitive than expected. This discrepancy is discussed both in terms of the conditions of pupae and in terms of mine structure. The results indicate that broadbandedness of vibrations produced by hunting parasitoids during ovipositor insertion into the mine may be a major criterion used by leafminers to perceive parasitoid presence and to escape ovipositor stings. PMID- 12770465 TI - Comparison of Drosophila cytochrome P450 metabolism of natural and model substrates. AB - Metabolism of some insecticides and toxic natural plant compounds is known to involve cytochrome P450 enzymes. Correlations between insecticide resistance and deethylation of the model substrate, 7-ethoxycoumarin, have prompted its use in screens for potentially resistant insect populations. The applicability of this model substrate as an indicator of the enzyme activities and inductive responsiveness of cytochrome P450 isoforms involved in the metabolism of carnegine was investigated. This toxic isoquinoline alkaloid is found in the host plants of some species of cactophilic Drosophila. The results show that the ethoxycoumarin (ECOD) assay does not accurately predict carnegine metabolism either quantitatively or with respect to the overall pattern of activity. Therefore, the ECOD assay may be as isozyme-specific in insects as has already been demonstrated in mammals and its use as an indicator of general P450 activity is questionable. PMID- 12770466 TI - The pathway of ammonia assimilation in the silkworm, Bombyx mori. AB - Ammonia can easily be assimilated into amino acids and used for silk-protein synthesis in the silkworm, Bombyx mori. To determine the metabolic pathway of ammonia assimilation, silkworm larvae were injected with methionine sulfoximine (MS), a specific inhibitor of glutamine synthetase (GS). Activity of GS in the fat body 2h after treatment with 400&mgr;g MS decreased to less than 10% of the control activity, whereas MS had no effect on the activity of glutamate dehydrogenase (GDH), another enzyme which could possibly be responsible for ammonia assimilation. Glutamine concentration in the hemolymph rapidly decreased after MS treatment, while the ammonia level in the hemolymph sharply increased. Glutamine concentration in the hemolymph 4h after injection decreased with increasing doses of MS, whereas ammonia concentration increased in proportion to the MS dose. MS strongly blocked the incorporation of (15)N label into silk protein in larvae injected with (15)N ammonia acetate, while it slightly inhibited the incorporation of (15)N-amide glutamine into silk-protein. These results suggest that ammonia is mainly assimilated into glutamine via the action of GS and then converted into other amino acids for silk-protein synthesis and that GDH does not play a major role in ammonia assimilation in B. mori. PMID- 12770467 TI - The effect of flash duration and flash shape on entrainment in Pteroptyx malaccae, a synchronic Southeast Asian firefly. AB - Southeast Asian synchronic fireflies respond to stimulus flashes by phase shifting their endogenous oscillator. This is called 'flash entrainment'. The releasers for entrainment were studied by changing stimulus flash shape and duration in Pteroptyx malaccae. Stimulus flash shapes and durations were synthesized digitally and delivered by a field-portable computer system. The computer also recorded male firefly flashes that were detected with a photometer. We found that the type of entrainment and its magnitude was influenced by the duration of the entrainment flash and by its shape. PMID- 12770468 TI - Effects of starvation on body composition and cold tolerance in the collembolan Orchesella cincta and the isopod Porcellio scaber. AB - The effects of long-term starvation on the body composition of the isopod Porcellio scaber (Latreille) and the collembolan Orchesella cincta (L.) were studied, by determining the body composition in starved and fed animals. A period under summer conditions (19 degrees C, 75% RH and L/D 16/8 photoperiod), was followed by a period under winter conditions (5 degrees C, 75% RH and LD 6/18 photoperiod). O. cincta was held under summer conditions for 3weeks, during which its protein and lipid content decreased, while its water content increased. In P. scaber, the same occurred during the 6weeks they were kept under summer conditions. During subsequent weeks under winter conditions, changes in cold tolerance of the animals were investigated. Cold tolerance and haemolymph osmolality were measured once a week. Starved animals had lower cold tolerance than fed ones. For P. scaber a decreased haemolymph osmolality was found in starved animals compared to fed ones. This is assumed to be caused by a combination of the consumption of carbohydrates out of the haemolymph and of protein reserves and the accumulation of body water. O. cincta appeared to be capable of osmoregulation, as haemolymph osmolality did not differ between starved and fed animals, despite differences in body water content. Decreased cold tolerance in starved animals of both species may be caused by increased water content or, more probably, by the decrease in reserves needed to produce cryoprotective substances. PMID- 12770469 TI - The influence of a static, homogenous magnetic field (B=320mT) on extracardiac pulsations of Tenebrio molitor pupae (Coleoptera: Tenebrionidae). AB - While investigating and describing interactions among living organisms and magnetic fields (MFs) it is imperative to lay great emphasis on independent reproducibility of published experimental results. Mutual confrontation of existing theoretical models with reliable data obtained under comparable conditions can aid gradual mapping of this hitherto badly organized and understood discipline of biology. The objective of our experiment, based on analysing extracardiac pulsations of the pupae of Tenebrio molitor under the influence of a MF, was to verify published data on allegedly accelerated development induced by a MF employing a different procedure. The obtained data are in agreement with a hypothesis of increased pupal metabolism during the period of MF activity. Furthermore, some dependence on the age of the pupae cannot be ruled out. PMID- 12770470 TI - Natriuretic and depolarizing effects of a stable fly (Stomoxys calcitrans) factor on Malpighian tubules. AB - A two-step HPLC purification procedure resulted in a factor from the stable fly that depolarizes the lumen-negative transepithelial voltage (V(t)) of the adult stable fly Malpighian tubule. When applied to tubules of the female mosquito, Aedes aegypti, this factor partially mimics the electrophysiological actions of the mosquito natriuretic factor (MNF). It also selectively increases active transepithelial Na transport by the mosquito Malpighian tubule. The blood meal causes a transient increase in hemolymph Na and Cl contents and hemolymph volume during the course of the 24-h post-feeding period. The level of a factor that is immunologically cross-reactive with the human atrial natriuretic peptide (ANP) increases more than 6-fold within 6h following a blood meal by the stable fly. The temporal pattern of the levels of the ANP-immunoreactive factor closely parallels the blood meal-induced rise and subsequent fall in hemolymph NaCl content and hemolymph volume, suggesting a functional correlation between the ANP immunoreactive factor and the rate of NaCl and fluid loss from the hemolymph. PMID- 12770471 TI - Occurrence of apoptosis in serosa of Periplaneta americana l. (Blattaria: blattidae): ultrastructural and biochemical features. AB - In 16-17-day-old embryos of Periplaneta americana, the amnion-serosa penetrates the cavity of the middle intestine, where it forms a cluster of compressed roundish cells. We demonstrated that these cells degenerate throughout apoptosis. The programmed cell death revealed by morphological and biochemical approaches showed all the apoptotic steps: chromatin fragmentation and pyknosis, cytoplasm condensation, karyorrhexis, cytoplasm cleavage. Nevertheless, some ultrastructural peculiarities (atypical heterochromatin arrangement, appearance of nuclear envelope protrusions, absence of nucleolar structures) suggest that the apoptotic expression partially depends on the biological situation (type of organism and inducing factors) in which the programmed cell death takes place. The presence of histiocytic cells internalizing cell debris, of apoptotic and non apototic derivation, may be correlated with the importance of recycling substances useful for embryo growth. PMID- 12770472 TI - Surfactants in the gut fluids of Porcellio scaber (Isopoda: Oniscidea), and their interactions with phenolics. AB - Fluids from the gut lumen of Porcellio scaber showed significantly reduced surface tension compared to a buffer solution. Tests with several dilutions indicated that the concentration of the surface active substances (surfactants) was about 80-fold higher than the 'critical micelle concentration'. Phenolics, e.g. gallotannins, when ingested in the diet increased the surface tension of the gut fluid, indicating reduced concentrations of free surfactants. The significance of gut surfactants in P. scaber, their role in digestive processes, and their interaction with tannins in this saprophagous soil arthropod are discussed. PMID- 12770473 TI - Contributions to Physiology of the Antenna-Heart in Periplaneta americana (L.) (Blattodea: Blattidae). AB - An accessory pulsatile organ of an open circulatory system in insects supplying the antennae with haemolymph was investigated. The rhythm of this so-called antenna-heart is generated by a myogenic automatism and can be neuronally influenced via the nervus cardioantennalis.The action potentials of the muscle fibres show typical pre-depolarization and mostly no overshoot. A specific membrane resistance (R(m)) of about 660Omegacm(-2) was calculated for the fibres. Some electrical coupling between the muscle fibres is presumed for synchronization of any myogenically triggered heart beat which could actually be proved experimentally by current injection in the antenna-heart. However, intercalated disks or gap junctions could not be found. Nevertheless, a good coupling factor (U(2)/U(1)) between all fibres was demonstrated by parallel recordings and can be well described by a conductance model according to fibre topology. PMID- 12770474 TI - Haemolymph proteins of larvae of Galleria mellonella detoxify endotoxins of the insect pathogenic bacteria Xenorhabdus nematophilus (Enterobacteriaceae). AB - Haemolymph of non-vaccinated Galleria mellonella larvae contains two proteins, LBP-1 (17.2kDa) and LBP-2 (26.0kDa) that:bond to the surfaces of the insect pathogenic bacteria, Xenorhabdus nematophilus;prevented lipid A-binding dye attaching to the lipid A of X. nematophilus endotoxin; andreduced endotoxin activity on the haemocytes.Protein LBP-1 also blocked the inhibition of prophenoloxidase activation by the endotoxins. It is proposed that proteins LBP-1 and LBP-2 are part of the containment responses of the insects to bacteria. PMID- 12770475 TI - Effects of alkali metal cations on the labellar water receptor cell of the fleshfly, Boettcherisca peregrina. AB - Dose-response relationships of the labellar water receptor cells of the fleshfly, Boettcherisca peregrina, in response to alkali-metal ions were studied electrophysiologically. When cation concentration was raised, the frequency of spikes from the water receptor cells was decreased. We analyzed the data under the assumption that channels on the water receptor membrane open when stimulated with water and are closed by alkali cations or nonelectrolytes. Dose-response relationships of the water receptor for NaCl, KCl and LiCl were analyzed using the Hill equation. The Hill coefficients and the activities of salts eliciting the half maximal response differed considerably among these salts. It is concluded that the effectiveness of salts on the inhibition of the water receptor cell differs among alkali metal ions, and that alkali cations interact cooperatively with the receptor membrane. PMID- 12770476 TI - Diet and carbohydrate digestion in the yellow-spotted longicorn beetle Psacothea hilaris. AB - Larval and adult Psacothea hilaris feed on mulberry wood and leaves, respectively. High levels of endogenous activity against the major dietary carbohydrates, cellulose, hemicellulose, starch and soluble sugars were secreted in the gut of larvae and adults. Activity against pectin was also high and multiple polygalacturonase (EC 3.2.1.15) components were secreted in the gut of larvae. One glycanase component, beta-EG1, which was primarily an endo-beta-1,4 glucanase (EC 3.2.1.4) and another, beta-EG2, which was mostly an endo-beta-1,4 xylanase (EC 3.2.1.8), were also secreted, while at least four additional components hydrolysed laminarin, lichenin and crystalline cellulose. The beta glycosidase component beta-GD1 was associated with most of the beta-mannosidase (EC 3.2.1.25) and beta-xylosidase (EC 3.2.1.37) activity secreted in the gut of larvae, while another, beta-GD2, was a beta-glucosidase (EC 3.2.1.21), the activity of which was directed against cellobiose and other beta-linked disaccharides, and a beta-fucosidase (EC 3.2.1.38). A beta-galactosidase (EC 3.2.1.23), which did not hydrolyse lactose, was also secreted, as were distinct beta-N-acetylhexosaminidase (EC 3.2.1.52), trehalase (EC 3.2.1.28), alpha-L arabinosidase (EC 3.2.1.55), alpha-galactosidase (EC 3.2.1.22) and a minimum of four alpha-glucosidase (EC 3.2.1.20) components, one of which was also likely to be associated with a peak of alpha-mannosidase (EC 3.2.1.24) activity. The alpha glucosidase components varied in their specificity for alpha-linked disaccharides, but none was active against sucrose, which was hydrolysed by a beta-fructofuranosidase (EC 3.2.1.26) component. Overall average levels of activity in larvae were twice those of adults, but the secretion of individual carbohydrases in both was not regulated in response to the relative abundance of particular carbohydrate components in their respective diets. PMID- 12770477 TI - Removing antennae and maxillae has little effect on feeding on normal host plants by two species of caterpillar. AB - Models of feeding regulation postulate that chemosensory information from available food both initiates and maintains feeding. However, we find that removing antennae and maxillae (AM) from Manduca sexta and Diacrisia virginica larvae has little effect on amounts eaten, patterns of feeding, and the microstructure (each bite and pause) of feeding, as quickly as two days after the operation. However, there was a small change in the microstructure of feeding of AM D. virginica. Bite frequency of AM D. virginica was significantly lower than for controls when larvae began meals without exploring the food first. Exploring was also followed by longer chewing bouts. Acute effects of the ablation were tested using only Manduca. Six hours after the operation larvae ate less than most controls by eating fewer meals and by biting more slowly. Unilateral ablates also ate less 6h post-operatively, by eating fewer meals; their bite frequency was not low. These observations suggest that chemosensory input affects bite frequency but not other aspects of feeding. As quickly as 24h post-operatively other sensory input to the CNS of AM larvae may compensate for the reduction in chemosensory information, but overall, chemosensory input appears relatively unimportant in non-choice situations. PMID- 12770478 TI - Cuticular hydrocarbons and their role in copulatory behavior in Phormia regina (Meigen). AB - The cuticular hydrocarbons from adult Phormia regina (Meigen) were characterized by gas chromatography-mass spectrometry. Both sexes had similar components in nearly identical quantities, consisting of complex mixtures of saturated n-, monomethyl- and dimethylalkanes from 23 to 33 total carbons. Although no diet-, age-, or sex-specific differences were observed, cuticular hydrocarbons were shown to be involved in copulatory behavior. Hydrocarbon profiles of wild, compared to laboratory reared flies, showed no major differences. Behaviorally, males responded the same to dead decoys of either sex. Removal of the hydrocarbons, using hexane, from either male or female decoys, did not affect the number of mating strikes, but markedly reduced the number of copulatory attempts and the amount of time males spent mounted on either decoy. House fly, Musca domestica L., males when paired with a female M. domestica decoy produced copulatory attempts: whereas, when P. regina males were placed with M. domestica female decoys, there were no copulatory attempts. It is concluded that the cuticular hydrocarbons of P. regina function as species-specific but not sex specific mating cues and elicit species-specific copulatory behavior in males. PMID- 12770479 TI - The effects of azadirachtin on the testes of the desert locust, Schistocerca gregaria (Forskal). AB - Testes removed from Schistocerca gregaria, injected with 5 or 7&mgr;g azadirachting(-1) body wt on day 0-2 of the Vth instar and killed by decapitation 20days after treatment, showed significant reductions between the widths, lengths and volumes of the treated groups, compared to control insects. Under cytological examination these testes, from insects exhibiting classical signs of azadirachtin poisoning, showed arrested spermatogenic meiosis at Metaphase I. PMID- 12770480 TI - Control of the eupyrene-apyrene sperm dimorphism in Lepidoptera. AB - Lepidoptera males bear concomitantly nucleate (eupyrene) and anucleate (apyrene) spermatozoa. Both kinds of spermatozoa reach the spermatheca of inseminated females but only the eupyrene ones fertilize the eggs. The functions of the apyrene spermatozoa are still uncertain. Eupyrene spermatogenesis is regular and highly sensitive to genetic and experimental manipulations while apyrene spermatogenesis is irregular and withstands these manipulations. Both kinds of spermatozoa derive from the same kind of bipotential spermatocytes. The shift of spermatocyte commitment from eupyrene to apyrene spermatogenesis is induced by a haemolymph factor that becomes active just before or after pupation, depending on species. Accordingly, eupyrene spermatogenesis starts during larval instars and stops after pupation while apyrene spermatogenesis begins just before or after pupation, depending on the species, and persists in the imago. The shift is related to shortening of meiotic prophases and blocking synthesis of a meiotic lysine-rich protein fraction in apyrene cells. From spermatogonia proliferation to early spermatocytes, spermatogenesis is a quasi-independent process. Afterwards, it becomes discontinuous and is punctuated by predetermined stations. Progress to a subsequent station is an 'all or none' phenomenon, regulated by cues linked to fluctuations of the main morphogenetic hormones titers. In absence of a particular cue, the cells stop advancing towards the next station and eventually degenerate. PMID- 12770481 TI - Change in reductase activity is responsible for senescent decline in sex pheromone titre in the lightbrown apple moth, Epiphyas postvittana (Walker). AB - Sex pheromone titre in the tortricid moth Epiphyas postvittana follows a pattern commonly observed in other species of moths: an increase to a peak some time after eclosion (2-3days), and then a slow decline as the female ages. Previous work has shown that this decline is not regulated by the pheromone biosynthesis activating neuropeptide PBAN. Using in vivo and in vitro enzyme assays, and fatty acid methyl ester (FAME) analyses of pheromone precursors in the gland, we have investigated this senescent decline in pheromone titre. The enzyme assays have shown that in older females the fatty acid reductase and fatty acid synthesis enzyme systems decrease in activity (relative to younger females), whereas other enzyme systems involved in pheromone biosynthesis, including limited beta oxidation (2-carbon chain-shortening), (E)-11-desaturation, and acetylation (by an acetyl transferase) remain unchanged in their activity. Of the two enzymatic processes involved, the more important one contributing to the decline appears to be the fatty acid reductase. This is consistent with FAME analyses of pheromone glands in old and young females, which show little difference in levels of saturated FAME, but a significant increase in the level of the putative precursor, (E)-11-tetradecenoate, of the sex pheromone component (E)-11 tetradecenyl acetate. Thus, this decline in fatty acid reductase activity results in a buildup of the precursor as the female ages. The near ubiquity of fatty acid reductases in moth sex pheromone systems suggests that this may be a common mechanism for the senescent decline of sex pheromone titre in moths. PMID- 12770482 TI - Neuroendocrine control of diapause hormone secretion in the silkworm, Bombyx mori. AB - To clarify the control mechanism of diapause hormone (DH) secretion in the silkworm Bombyx mori a series of anatomical and pharmacological experiments were carried out. The arrangement of 'diapause' and 'non-diapause' eggs in the ovarioles of the moths was determined by the coloration method to estimate the accumulation of 3-hydroxykynurenine in the eggs. The females destined to lay non diapause eggs (non-diapause producers) had diapause eggs in their ovaries if their subesophageal ganglions (Sg) had been surgically removed at 2days after larval-pupal ecdysis or later. In contrast when the surgical extirpation extended to the brain and the corpora cardiaca (CC)-corpora allata (CA) complex in addition to the Sg, the non-diapause producers had no diapause eggs. When the Sg was removed from the females destined to lay diapause eggs (diapause-producers), diapause eggs appeared in response to the treatment at 2days after larval-pupal ecdysis, but the appearance of diapause eggs was delayed by 2days when the brain CC-CA complex was included among the organs removed. These observations suggested that DH is produced in Sg and transferred to the CC-CA complex, and that the secretion of DH from the complex is suppressed in non-diapause producers. The pattern of diapause and non-diapause eggs induced by the transection of the subesophageal connective in diapause and non-diapause producers suggested a regenerative and secretory capacity of the neurosecretory cells after the operation. The appearance of diapause eggs in non-diapause producers with transected protocerebrum of the brain confirmed that there was an inhibitory center in the protocerebrum. Changes in parts of the ovarioles containing diapause and non-diapause eggs with time of injection of gamma-aminobutyric acid (GABA) and picrotoxin suggested that a GABAergic inhibitory mechanism in DH secretion may be active in non-diapause producers but inactive in diapause producers throughout the pupal stage. PMID- 12770483 TI - Incorporation of fatty acids into cuticular hydrocarbons of male and female Drosophila melanogaster. AB - The biosynthesis of cuticular hydrocarbons was investigated in male and female Drosophila melanogaster (Canton-S strain), especially in those with a pheromonal role i.e. male 7-tricosene and female 7, 11- heptacosadiene. The incorporation of radioactivity was followed after topical application of (14)C-labelled myristic, palmitic and stearic acid and (3)H-labelled cis-vaccenic acid on one to ten day old flies. The incorporation levels into unsaturated hydrocarbons are similar in both sexes, depending markedly on the chain length of the saturated precursor, with a maximum level from myristic acid. Cis-vaccenic acid leads only to unsaturated compounds. With this precursor, there is an enhanced incorporation into female monoenes and dienes, maximum in two to three day old females. The total fatty acid composition shows the highest abundance of fatty acids with 16 carbon atoms and the presence of a major position for double bond, Delta9. Moreover, cis-vaccenic acid and 17-tetracosenoic acid are identified by GC-MS analysis. These data support an elongation-decarboxylation mechanism for the biosynthesis of D. melanogaster cuticular hydrocarbons. Its early steps for male monoenes and female monoenes and dienes might involve a Delta9 desaturase transforming palmitic acid into palmitoleic acid which would then be elongated into vaccenic acid, an important common precursor for all pheromones. PMID- 12770484 TI - Mating and hormonal triggers regulate accessory gland gene expression in male Drosophila. AB - Drosophila melanogaster males transfer accessory gland proteins, as part of their seminal fluid, to females during each mating. Since accessory gland proteins are important for male reproductive success, it is important that the male replenish the proteins he transferred during mating. Previous studies had shown that mating induces the resynthesis of accessory gland proteins, but since mating includes a set of stereotyped behavior patterns as well as the act of copulation, it was not known which aspect of the mating process induces accessory gland protein synthesis. By exposing males to females whose ovipositors had been sealed shut, we have shown that resynthesis of accessory gland proteins occurs only when seminal fluid is transferred to females. By applying juvenile hormone or 20 hydroxyecdysone topically to the cuticle of male flies, we showed that these hormones can act in vivo to stimulate the synthesis of accessory gland proteins to levels similar to those observed after mating. PMID- 12770485 TI - Activation of picrotoxin-resistant GABA receptors by GABA and related compounds induces modulation of cockroach dorsal paired median (DPM) neuron firing. AB - Activation of gamma-aminobutyric acid (GABA) receptors in insect dorsal paired median (DPM) neurons induced two types of response which appeared to be mediated by two different GABA receptor subtypes. When activated by bath application of GABA, one receptor subtype, insensitive to picrotoxin (PTX), mediated a drastic reduction in the firing frequency, leading to a blockade of the spontaneous electrical activity. These effects were accompanied by decreases in the amplitude and duration of the plateau action potential (AP) and the spike after hyperpolarization (AHP). In most cases, a slight depolarization of the resting membrane potential occurred. Bath application of the vertebrate GABA(B) receptor agonists 3-aminopropyl(methyl)phosphinic acid (SKF 97541) and 3 aminopropylphosphinic acid (CGA 147823/CGP 27492) induced similar responses. Another GABA receptor subtype, less sensitive to GABA, mediated a chloride dependent hyperpolarization that was suppressed by bath application of PTX. The approximate locations of these two GABA receptor subtypes were determined by local pressure microapplications of GABA and vertebrate GABAergic agonists. The PTX-sensitive receptors were located predominantly on the surface of the ganglion where the apical pole of the soma is situated, while the PTX-resistant receptors appeared to be located deeper within the ganglion.These results reveal the existence of two GABA receptor subtypes on the DPM neurons and provide evidence for a functional role for PTX-resistant GABA receptors in the regulation of spontaneous firing. PMID- 12770486 TI - Circadian rhythm of sperm release in the gypsy moth, Lymantria dispar: ultrastructural study of transepithelial penetration of sperm bundles. AB - Release of mature bundles of spermatozoa from the testis into the vas deferens is a critical but poorly understood step in male insect reproduction. In moths, the release of sperm bundles is controlled by a circadian clock which imposes a temporal gate on the daily exit of bundles through the terminal epithelium-a layer of specialized epithelial cells separating testis follicles from the vas deferens. The sequence of cellular events associated with the daily cycle of sperm release was investigated by scanning and transmission electron microscopy. In the hours preceding sperm release, there is a solid barrier between the testis and the vas deferens formed by the interdigitation of cytoplasmic processes of adjacent terminal epithelial cells. At the beginning of the sperm release cycle, sperm bundles protrude through this barrier while the terminal epithelial cells change their shape and position relative to the bundles. Subsequently, the cyst cells enveloping the sperm bundles break down and spermatozoa move out of the testis through the exit channels formed between the epithelial cells. Afterwards, cyst cell remnants and other cellular debris are released into the vas deferens lumen, and the epithelial barrier is reconstructed due to phagocytic activity of its cells. These data provide a foundation on which to build an understanding of the cellular mechanisms of clock-controlled sperm release in insects. PMID- 12770487 TI - Effects of the entomopathogenic fungus Metarhizium anisopliae and its secondary metabolites on morphology and cytoskeleton of plasmatocytes isolated from the greater wax moth, Galleria mellonella. AB - The effects of Metarhizium anisopliae infection and three different secondary metabolites released by the fungus, destruxin A and E and cytochalasin D, on the morphology and cytoskeleton of plasmatocytes of the greater wax moth Galleria mellonella were studied. Plasmatocytes isolated from M. anisopliae infected larvae exhibited impairment of attachment, spreading and cytoskeleton formation accompanied with the occurrence of blebbing and pycnotic nuclei. Plasmatocytes treated with destruxin in vitro exhibited similar morphological and cytoskeleton alterations. The corresponding effects were characterized by inhibition of attachment, spreading and filopodia formation as well as by impaired formation of actin filaments and microtubules. Cytochalasin was shown to affect plasmatocytes in vitro in a different manner than destruxin A and E. The results of our comparative study strongly suggested that the morphology and cytoskeleton alterations of plasmatocytes observed in M. anisopliae infected larvae were predominantly caused by destruxins released by the fungus during mycosis. Its mode of action is discussed with regard to present knowledge about its effects on target cells. PMID- 12770488 TI - Juvenile hormone regulates the expression of the gene encoding ceratotoxin a, an antibacterial peptide from the female reproductive accessory glands of the medfly Ceratitis capitata. AB - Ceratotoxin A is an antibacterial peptide produced by the reproductive female accessory glands of the medfly Ceratitis capitata. To investigate whether ceratotoxin A gene expression was affected by juvenile hormone, which has gonadotropic functions in adult insects, newly emerged female medflies were treated with precocene II, an antiallotropin compound capable of inhibiting juvenile hormone biosynthesis. Daily treatment of newly emerged flies with precocene II blocked ceratotoxin A gene expression in a dose-dependent manner. Ceratotoxin A gene expression could be recovered after withdrawl of precocene II treatment. Moreover, the effect of precocene II on ceratotoxin A gene expression could be countered by simultaneous treatment with methoprene, a juvenile hormone analogue. The effects of precocene II and methoprene treatments on the growth of both ovaries and accessory glands was also investigated. Our data suggest that ceratotoxin A gene expression is modulated by juvenile hormone. PMID- 12770489 TI - Impact of dietary allelochemicals on gypsy moth (Lymantria dispar) caterpillars: importance of midgut alkalinity. AB - Midgut pH of gypsy moth larvae was depressed artificially with buffered diet to examine the impact of alkalinity on the caterpillars' ability to tolerate a dietary polyphenol and a quinone. A 2x3 factorial design was used, with 2 levels of succinate buffer and 3 dietary amendments (tannic acid, juglone, or control). Development was monitored during the third and fourth instars, with consumption, food passage rates, midgut pH, and midgut redox potential (Eh) measured in the fourth instar. Diet buffering successfully depressed midgut pH to hypothetically suboptimal acidic levels without reductions in survivorship, but it did reduce larval growth and impede development. Buffering dramatically reduced survivorship of fourth instar larvae eating diets containing tannic acid or juglone. Growth increased on unbuffered diet amended with tannic acid, but not with juglone. Caterpillars passed food through the gut more slowly when feeding on buffered tannic acid diet or on unbuffered juglone diet. These results indicate that maintenance of midgut alkalinity is critical to tolerance of dietary tannic acid and juglone, and that these allelochemicals have very different activities in the caterpillar gut. PMID- 12770490 TI - Effects of juvenile hormone treatment on phase changes and pheromone production in the desert locust, Schistocerca gregaria (Forskal) (Orthoptera: Acrididae). AB - The roles of juvenile hormone III (JH III) on phase changes and pheromone production were examined in laboratory-reared gregarious desert locust, Schistocerca gregaria (Forskal). The hormone was applied to 5th instar nymphs and newly emerged adult locusts. Generally, the 5th instar nymphs exhibited a higher sensitivity to hormone treatments than the adults. Hormone applications inhibited pheromone production (as measured by the amounts of phenylacetonitrile released). In addition, JH III had a significant effect on the external colouration and absorbance ratios of the haemolymph pigments. It is concluded that the effects of exogenous JH III on gregarious locusts represent a shift towards the solitarious phase. PMID- 12770491 TI - Haemolymph osmoregulation and the fate of sodium and chloride during dehydration in terrestrial isopods. AB - Osmoregulation of the haemolymph during dehydration was investigated in a selection of temperate oniscidean isopods. Inulin tracer studies show that the haemolymph contributes approximately 69% of water losses in Porcellio scaber, significantly more than predicted from the volume of this compartment (42% of total water). Haemolymph osmolality increases linearly as a function of haemolymph dehydration but at a significantly lower rate than predicted from the change in haemolymph fluid volume. Similar results for Oniscus asellus show that both species display efficient osmoregulation until lethal dehydration. Osmoregulation is associated with significant hyporegulation of haemolymph sodium and chloride. These findings indicate that: (1) cell water is conserved at the expense of the haemolymph; and (2) haemolymph dehydration is associated with the removal of Na(+) and Cl(-) contributing to net osmoregulation. During dehydration, accumulations of both Na(+) and Cl(-) are seen in the hindgut, with significant accumulations of electrolytes also seen in the luminal fluid of the hepatopancreas. Low fluid volumes in the foregut and hindgut suggest macromolecular association as the most plausible mechanism of ion sequestration. Evidence refutes ion excretion and haemocyte sequestration as osmoregulatory mechanisms. Sequestration of Na(+) as urate salts, as shown for Periplaneta and generally assumed for other insects, is insignificant in isopods. PMID- 12770492 TI - Differentiation of dialects and courtship strategies in allopatric populations of Drosophila teissieri. AB - Intra-specific differentiation has been investigated between two geographically isolated populations of D. teissieri, one from Brazzaville (Congo) and the other from Silinda (Zimbabwe). Courtship songs were analyzed on 23 parameters. In addition, certain parameters related to sexual activity (five) and wing morphology (five) were also examined. As in other Drosophilidae, there are two types of courtship songs resulting from wing vibration: the love song, and the sine song. Love songs from Brazzaville are longer than those from Silinda (334ms vs. 216) with a tendency to smaller and more regular inter-pulse intervals (24.56ms vs. 25.5). The sine songs (SS) are about six times longer than the love songs in each population, with an adaptation of their duration according to the receptivity of the female in Brazzaville. All the LS temporal parameters, except the interpulse interval mean, are significantly different between the two populations. The Brazzaville flies show a greater acoustic activity and have higher mating success than the Silinda ones (53.3% vs. 27.7%). Their wings are slightly shorter and wider than those from Silinda. The role of acoustic parameters, sexual activity and wing physical constraints are discussed in the perspective of incipient speciation. PMID- 12770493 TI - Ecdysteroid titres in a tenebrionid beetle, Zophobas atratus: effects of grouping and isolation. AB - Metamorphosis in Zophobas atratus is dependent on isolation: when kept in grouped conditions, larvae undergo numerous supernumerary moults, growing in size, without pupating. This beetle thus represents an interesting model for the analysis of possible differences in the endocrine regulation of normal vs. supernumerary larval moults. In this study, the ecdysteroid titres have been analysed in this species, using enzyme immunoassay. The hormonal variations of larvae undergoing normal or supernumerary larval cycles were particularly examined, in either grouped or isolated conditions. Normal larval cycles presented very similar ecdysteroid variations in grouped as well as isolated conditions, showing a single hormonal peak (at about 1000pg/&mgr;l). Supplementary larval cycles, occurring in grouped conditions, also showed a similar single ecdysteroid peak, but after a longer period of basal levels. Isolation of such larvae triggered their larval-pupal transformation, which was characterized by more complex hormonal fluctuations, including a small ecdysteroid peak before the main one. Interestingly, the isolation of big larvae during a large part of their cycle induced this peculiar hormonal pattern synchronously, confirming the involvement of a complex neuroendocrine control between external conditions and ecdysteroid titres. PMID- 12770494 TI - Comparison of the response of midgut epithelial cells and cell lines from lepidopteran larvae to CryIA toxins from Bacillus thuringiensis. AB - The cytotoxic responses of midgut epithelial cells (MEC) from spruce budworm (SBW), gypsy moth (GM) and silkworm (SW) larvae were compared with the cytotoxic response of lepidopteran cell lines (SF-9, SE-1a, and CF-1) to CryIA toxins from Bacillus thuringiensis. The MEC from SBW, SW and GM had binding proteins for CryIA(a,b,c) toxins, whereas the lepidopteran cell lines had binding proteins for CryIA(c). Single MEC exposed to CryIA(a,b,c) toxins in a qualitative lawn assay were equally susceptible to the toxins with a threshold response at about 1ng. The cell lines were not susceptible to CryIA(a,b) toxins in the dose range tested, but had threshold responses for CryIA(c) of 3.4ng for SF-9, 50.2ng for SE 1a and 5.9ng for CF-1. In the quantitative Live/Dead assay, MEC were equally susceptible to CryIA(a,b,c) toxins with a threshold effect at about 1ng and a maximum effect at about 10ng. CF-1 was most sensitive to CryIA(c) with a threshold effect at 0.39ng and a maximal effect at about 1ng. In contrast, a 25 50 times greater dose of CryIA(a) or CryIA(b) was required to elicit a similar response as CryIA(c) for CF-1. SF-9 and SE-1a were most susceptible to CryIA(c) with a threshold effect observed at about 0.5ng and maximal effects at about 2ng. SF-9 cells have a threshold and maximum response to CryIA(a,b) of about 10ng and 20ng, respectively. SE-1a cells have a threshold and maximal response to CryIA(a,b) of 5ng and 10ng, respectively. Intact midgut epithelium exposed to CryIA(a,b,c) toxins had a threshold dose of 2ng for CryIA(b), 10-30ng for CryIA(a) and 2-30ng for CryIA(c). This study has shown that MEC are affected by a broader spectrum of toxins compared to the lepidopteran larvae and insect cell lines. PMID- 12770495 TI - Proteinase inhibitors from Nicotiana alata enhance plant resistance to insect pests. AB - The ornamental tobacco (Nicotiana alata) produces one 6-kDa chymotrypsin inhibitor and four 6-kDa trypsin inhibitors from a single 40.3-kDa precursor protein. Three different approaches have been used to assess the potential of these proteinase inhibitors (PIs) in insect control. The first was an in-vitro approach in which all five inhibitors, the single chymotrypsin inhibitor or three of the four trypsin inhibitors were tested for their ability to inhibit gut protease activity in insects from four orders. The second approach was to incorporate the N. alata PIs in the artificial diet of the native budworm (Helicoverpa punctigera) and the black field cricket (Teleogryllus commodus). H. punctigera larvae and T. commodus nymphs had a significant (P<0.01) reduction in growth after ingestion of the PI and were more lethargic than insects on the control diet. Several of the H. punctigera larvae also failed to complete moulting at the third or fourth instar. The third approach was to express the N. alata PIs in transgenic tobacco under the control of the 35S CaMV promoter. When H. punctigera larvae were fed tobacco leaves expressing the N. alata PIs at 0.2% soluble protein, significant (P<0.01) differences in mortality and/or growth rate were observed. PMID- 12770496 TI - Cold hardiness, supercooling ability and causes of low-temperature mortality in the soft tick, Argas reflexus, and the hard tick, Ixodes ricinus (Acari: Ixodoidea) from Central Europe. AB - Seasonal supercooling points (SCPs=temperature of crystallization) and cold hardiness were investigated in the indigenous hard tick, I. ricinus, and in A. reflexus, a soft tick introduced to Central Europe from the South. Both species proved to be freeze-susceptible as well as highly susceptible to inoculative freezing. None of the postembryonic developmental stages of either species showed any distinct seasonal pattern of SCP. Unexpectedly, the introduced A. reflexus exhibited a distinctly higher degree of cold hardiness in terms of lower lethal temperature (LT(50): 24h exposure) as well as lethal time (T(50): time of survival at -10.1 degrees C) than I. ricinus. Engorged I. ricinus larvae as well as engorged summer acclimatized A. reflexus larvae showed some mortality at temperatures well above the SCP. This mortality was generally expressed as a failure of the following stage to eclose properly. A 10-day cold acclimation at +3 degrees C eliminated that kind of mortality in summer acclimatized A. reflexus larvae, but not in I. ricinus larvae. It was frequently observed that freezing of ticks resulted-possibly via leakage from the midgut-in a subsequent reddish brown discoloration of the ticks after thawing. Taking into account that discoloration was an indication of previous freezing, it was concluded, that after long-term exposure (for >/=30 days) at -10.1 degrees C, a temperature well above the SCP, some tick mortality could be observed that was not caused by previous freezing. Weighing experiments clearly demonstrated, that the level of dehydration was not critical for survival of A. reflexus during long-term cold exposure, even at low RH. This indicates, that cold-related factors other than freezing and dehydration were detrimental to this species. PMID- 12770497 TI - Dietary regulation of serine proteinases that are resistant to serine proteinase inhibitors. AB - Ingestion of Kunitz soybean trypsin inhibitor (STI) by larval Helicoverpa zea, Agrotis ipsilon, and Trichoplusia ni extended the retention time of food in the digestive tract and increased the level of activity of proteolytic enzymes that were not susceptible to inhibition by STI. The level of enhancement of activity of STI-resistant (STI-R) enzyme(s) was directly influenced by the dosage and timing of exposure to STI. However, not all proteinase inhibitors (PIs) enhanced the level of proteinase inhibitor resistant (PI-R) enzymes, even if those PIs inhibited a significant proportion of enzyme activity. These findings suggest that a complex system may be responsible for the regulation of proteolytic enzymes in the midgut of larval Lepidoptera, and one hypothesis for this regulation is proposed. PMID- 12770498 TI - Development changes in juvenile hormone and juvenile hormone acid titers in the hemolymph and in-vitro juvenile hormone synthesis by corpora allata of the silkworm, Bombyx mori. AB - A simple method was developed to quantify hemolymph juvenile hormone (JH) and JH acid in hemolymph extracts from Bombyx mori with an established radioimmunoassay (RIA) for JH I. When various organic solvent extracts of hemolymph were assayed by RIA, levels of non-specific binding of the labeled ligand in the assay were determined to be greater than 50% of the maximum amount of the label bound by the antiserum. When hemolymph was diluted with methanol:water:8.4N ammonium hydroxide (10:9:1) and extracted with isooctane, non-specific binding was only 50% higher than control levels obtained with the assay buffer alone. The organic phase contained only JH and aqueous phase, JH acid. Consequently, this extraction method was used to prepare samples for RIA and enabled the separate measurement of JH and JH acid in hemolymph. With this method, changes in the hemolymph titers of JH and JH acid were determined from the third instar through early pupal stage of Bombyx mori. Changes in the in vitro secretory activity of corpora allata and brain-corpora cardiaca-corpora allata complexes from fifth instar larvae were also determined by using JH I RIA of the incubation medium. PMID- 12770499 TI - Cloning and expression of the ccdA-associated thiol-disulfide oxidoreductase (catA) gene from Brevibacillus choshinensis: stimulation of human epidermal growth factor production. AB - Brevibacillus choshinensis (Bacillus brevis) is a protein-hyperproducing bacterium with a useful host-vector system for the production of recombinant proteins. Here, we cloned the ccdA-catA (cmacr;cdA associated thioredoxin-like tmacr;hiol-disulfide oxidoreductase) locus of B. choshinensis HPD31-S5. CatA protein (molecular weight, 19664) contains a thioredoxin-like motif, Cys-Gly-Pro Cys. It was successfully expressed in B. choshinensis extracellularly ( approximately 100 microg x ml(-1) culture) using the secretion vector pNCMO2, and in Escherichia coli intracellularly ( approximately 350 microg x ml(-1) culture) with an amino-terminal His-tag. Both recombinant proteins showed thiol-disulfide oxidoreductase activity. Incubation of non-native human epidermal growth factor (hEGF) containing incorrect disulfide bonds with B. choshinensis cells secreting CatA protein resulted in the stimulation of the conversion of non-native hEGF to the native form. Furthermore, co-expression of CatA protein with recombinant hEGF in the B. choshinensis production system increased the yield of native hEGF. PMID- 12770500 TI - High level expression of the bioactive human interleukin-10 in milk of transgenic mice. AB - Human interleukin-10 (hIL-10) has wide spectrum of anti-inflammatory activities and has shown a potential to be used for treatment of inflammatory or immune illness. In this study, transgenic mice that over-express human interleukin-10 (IL-10) in their milk were generated using a bovine beta-casein/human IL-10 hybrid gene. After cloning of the IL-10 gene, a 22 kb hybrid gene was constructed by linking a 10 kb promoter sequence of the bovine beta-casein gene to the cloned 12 kb IL-10 gene. In six of the eight transgenic mice, the transgene RNA was expressed only in the mammary gland and in the other two mice, it was also slightly expressed in the lung. The highest human IL-10 level in milk was 1620 microg x ml(-1). Notably, transgenes in all the eight transgenic mice were expressed regardless of the integration site even though no correlation was shown between the copy numbers of the transgene and expression level. These results suggest that the genomic sequence of the human IL-10 gene can induce the IL-10 expression at high levels under the control of the bovine beta-casein promoter. PMID- 12770501 TI - Analysis of docosahexaenoic acid biosynthesis in Crypthecodinium cohnii by 13C labelling and desaturase inhibitor experiments. AB - The lipids of the heterotrophic microalga Crypthecodinium cohnii contain the omega-3 polyunsaturated fatty acid (PUFA) and docosahexaenoic acid (22:6) to a level of over 30%. The pathway of 22:6 synthesis in C. cohnii is unknown. The ability of C. cohnii to use 13C-labelled externally supplied precursor molecules for 22:6 biosynthesis was tested by 13C NMR analysis. Furthermore, the presence of desaturases (typical for aerobic PUFA synthesis) was studied by the addition of specific desaturase inhibitors in the growth medium. The addition of 1-(13)C acetate or 1-(13)C butyrate in the growth medium resulted in 22:6 with only the odd carbon atoms enriched. Apparently, two-carbon units were used as building blocks for 22:6 synthesis and butyrate was first split into two-carbon units prior to incorporation in 22:6. When 1-(13)C oleic acid was added to the growth medium, 1-(13)C oleic acid was incorporated into the lipids of C. cohnii but was not used as a precursor for the synthesis of 22:6. Specific desaturase inhibitors (norflurazon and propyl gallate) inhibited lipid accumulation in C. cohnii. The fatty acid profile, however, was not altered. In contrast, in the arachidonic acid-producing fungus, Mortierella alpina, these inhibitors not only decreased the lipid content but also altered the fatty acid profile. Our results can be explained by the presence of three tightly regulated separate systems for the fatty acid production by C. cohnii, namely for (1). the biosynthesis of saturated fatty acids, (2). the conversion of saturated fatty acids to monounsaturated fatty acids and (3). the de novo synthesis of 22:6 with desaturases involved. PMID- 12770502 TI - The effect of medium composition on the structure and physical state of sophorolipids produced by Candida bombicola ATCC 22214. AB - Candida bombicola was grown using a variety of lipophilic carbon substrates. Most of the hydrocarbon and carboxylic acid substrates resulted in a mixture of sophorolipids consisting of free acids and the more desirable lactones. The ratio of diacylated lactone to free acid in these mixtures was a maximum when produced using hexadecane and heptadecane. All of the other lipophilic substrates resulted in significant amounts of free acids being produced. These lactone products were unique in that they precipitated as crystals, which were easily separated from the culture medium. All of the other products were isolated as oils as is usually reported in the literature. Finally, the amounts of these crystals recovered were significantly higher than those observed for any of the oily products. It was possible to determine the degree of direct incorporation of the lipophilic substrates into the sophorolipids for a homologous series of alkanes. The amount of direct incorporation increased with increasing chain length to a maximum for pentadecane, hexadecane and heptadecane. As the length of the alkane substrate increased further, the amount of direct incorporation then decreased until there was no apparent incorporation for eicosane. PMID- 12770503 TI - Synthesis of mumps virus nucleocapsid protein in yeast Pichia pastoris. AB - The expression of mumps virus nucleocapsid protein in yeast Pichia pastoris was investigated. Viral nucleocapsid proteins usually elicit a strong long-term humoral immune response in patients and experimental animals. Therefore, the detection of antibodies specific to mumps virus nucleoprotein can play an important role in immunoassays for mumps diagnosis. For producing a high-level of recombinant mumps virus nucleoprotein the expression system of yeast P. pastoris was employed. The recombinant nucleocapsid protein was purified by cesium chloride ultracentrifugation of yeast lysates. Electron microscopy of the purified recombinant nucleocapsid protein revealed a herring-bone structure similar to the one discovered in mammalian cells infected with mumps virus. The yield of purified nucleocapsid-like particles from P. pastoris constituted 2.1 mg per 1 g of wet biomass and was considerably higher in comparison to the other expression systems. PMID- 12770504 TI - The putative transcriptional repressor McbR, member of the TetR-family, is involved in the regulation of the metabolic network directing the synthesis of sulfur containing amino acids in Corynebacterium glutamicum. AB - In order to isolate transcriptional regulatory proteins involved in L-methionine dependent repression in Corynebacterium glutamicum, proteins binding to the putative promoter region upstream of the metY gene were isolated by DNA affinity chromatography. One of the isolated proteins was identified as a putative transcriptional repressor of the TetR-family by a mass spectrometry fingerprint technique based on the complete C. glutamicum genome sequence. The respective gene, designated mcbR, was deleted in the mutant strain C. glutamicum DR1. Using 2D-PAGE, the protein contents of the C. glutamicum wild type and the mutant strain DR1 grown in media with or without L-methionine supplementation were compared and a set of six proteins was identified. Their abundance was drastically enhanced in the mutant strain and no longer influenced by L methionine added to the growth medium. The corresponding genes were identified by mass spectrometry fingerprint analysis. They included metY encoding O-acetyl-L homoserine sulfhydrylase, metK encoding S-adenosyl-methionine synthethase, hom encoding homoserine dehydrogenase, cysK encoding L-cysteine synthase, cysI encoding an NADPH dependant sulfite reductase, and ssuD encoding an alkanesulfonate monooxygenase. Evidently, the putative transcriptional repressor McbR is involved in the regulation of the metabolic network directing the synthesis of L-methionine in C. glutamicum. The C. glutamicum mcbR mutant can be considered to represent a first step in the construction of an L-methionine production strain. PMID- 12770505 TI - Effects of bacterial treatments on wood extractives. AB - Bacterial strains were isolated from spruce wood chips and their ability to reduce the content of wood extractives was studied. Strains were screened by cultivation on liquid media containing wood extractives as the major nutrient. Some bacterial species could decrease remarkably the amount of extractives in the liquid media and reduced the amount of triglycerides, steryl esters and total extractives by 100, 20 and 39%, respectively. Spruce wood chips were treated in controlled conditions with selected bacteria to test their effects on the chips. All the bacteria grew well on wood chips. The effect of bacterial metabolism on wood extractives was significant. Bacterial treatments reduced the amount of lipophilic extractives by 16-38% in 1 week of treatment and up to 67% in 2 weeks. The most efficient strain removed 90, 66 and 50% of triglycerides, steryl esters and resin acids, respectively, in 2 weeks. These results indicate that bacteria may be promising agents for the removal of extractives for improved pulping and papermaking processes. PMID- 12770506 TI - Submerged cultivation of Ganoderma lucidum biomass and immunostimulatory effects of fungal polysaccharides. AB - Original Ganoderma lucidum strain MZKI G97 isolated from Slovenian forests was cultivated in a liquid substrate based on potato dextrose and olive oil. The influences of inoculum and oxygen partial pressure in batch and fed batch cultivation in a 10-l laboratory stirred tank reactor were studied. Fungal biomass was found to be oxygen and shear sensible. Using a 17% (wet weight) 6 days old vegetative inoculum, 9.6 g l(-1) of dry biomass in batch cultivation and 15.2 g l(-1) in fed batch process were obtained. Extracellular (9.6 g l(-1)) and intracellular (6.3 g l(-1)) polysaccharide fractions were isolated. Extracellular polysaccharide fraction and four intracellular polysaccharide fractions were obtained. Polysaccharides were further separated by ion-exchange, gel and affinity chromatography. The isolated polysaccharides were mainly beta-D glucanes. Immunostimulatory effects of isolates were tested on induction of cytokine (tumour necrosis factor alpha (TNF-alpha) and interferon gamma (IFN gamma)) synthesis in primary cultures of human peripheral blood mononuclear cells (PBMC) isolated from a buffy coat. The TNF-alpha inducing activity is comparable with romurtide, which has been used as a supporting therapy in cancer patients treated with radiotherapy and/or chemotherapy. PMID- 12770507 TI - Monitoring batch fermentations with an electronic tongue. AB - An electronic tongue comprising 21 potentiometric chemical sensors with pattern recognition tools was used for the rapid off-line monitoring of batch Escherichia coli fermentations. The electronic tongue was capable of monitoring the changes in the media composition as the fermentation progressed, and could correlate this with an increase in biomass. The electronic tongue was also able to monitor the increase in organic acids, especially acetic acid, throughout the fermentation. This technique clearly shows promise as a rapid tool for fermentation monitoring. PMID- 12770508 TI - Emerging problems in the pharmacology of migraine: interactions between triptans and drugs for prophylaxis. AB - Patients suffering from migraine take drugs for many years in order either to relieve or to prevent recurrent migraine attacks. When two or more drugs are co administered, there is always the possibility of drug-drug interaction. Interactions can be either kinetic or dynamic. The former are the most frequent ones. Mechanisms of kinetic interaction can be different, but the most common are represented by the induction or inhibition of enzymes of the cytochromes p450 (CYP) system. This system plays an important role in the disposition of a large number of drugs, including those used for migraine. This review examines the interactions between triptans-the most effective drugs for the therapy of migraine attacks-and drugs for migraine prophylactic treatment. PMID- 12770509 TI - Effects of Daflon on oxidative stress induced by hindlimb ischemia/reperfusion. AB - The objective of this study was to investigate the effects of Daflon 500 mg on tissue damage in kidney after ischemia/reperfusion hindlimb, by assessing blood biochemical assay and histopathological analysis. Rats were given Daflon 80 mg x kg(-1) x day(-1) for 10 days. On 11th day of treatment, 4h ischemia followed by 4 h reperfusion period was performed on right hind limb of the rats. Control groups were given only arabic gum and were subjected to same ischemia/reperfusion period. At the end of reperfusion period, erythrocyte superoxide dismutase, Na(+) K(+) ATPase and reduced glutathione levels were increased in the rats erythrocytes in Daflon group (P<0.01, for all). On the other hand, serum myeloperoxidase and malondialdehyde levels were significantly lower in the Daflon received rats (P<0.01, for all). Histopathological studies demonstrated that, there was a prominent tubulointerstitial injury with loss of brush border and this degeneration was accompanied by segmental glomerular degeneration also for both control and Daflon group. Daflon-received group animals displayed significantly less periglomerular and perivascular leukocytic infiltration (P=0.015). These overall results suggest that Daflon contributes renal protection from hind limb ischemia/reperfusion injury in some degree, by decreasing systemic oxidative stress. PMID- 12770510 TI - Effects of clonidine on the experimental hypertension by abdominal aortic coarctation in rats. AB - Cardiovascular baroreflex mechanisms and sympathetic tone could be involved in the arterial hypertension by coarctation of abdominal aorta artery (CoA). The present work analyzes the effect on the arterial pressure and heart rate (HR) of the clonidine, an alpha(2)-adrenergic central acting antihypertensive agent, after intravenous (i.v.), intracerebroventricular (i.c.v.) and intrathecal (i.t.) administration in rats anesthetized with pentobarbital (40 mg/kg i.p.).Wistar rats of both sexes (240-270 g) were used to the 7 days of the CoA or a sham operation (SO). Values of mean arterial pressure (MAP) and of HR were calculated from intraarterial recordings of blood pressure. The MAP of the CoA rats (161.5+/ 5.3 mmHg, n=20) was significantly higher (P<0.01) than that of the SO rats (101.6+/-3.3 mmHg, n=20). The i.v. injection of clonidine (3-30 microg/kg) produced an increase of blood pressure in the rats SO and in the CoA animals, followed by a fall of arterial pressure in both groups of rats. Clonidine showed a small pressor effect but also a great depressor action in the hypertensive rats. Except for with the dose of 10 microg/kg, differences in cardiac response to clonidine were not seen in both groups of rats. Injection of clonidine by the i.c.v. via (10 microg) like by the i.t. (3 microg) also produced a greater fallen of the MAP in the hypertensive rats than in the controls SO animals. In conclusion, these hypertensive animals would be sensitive to the antihypertensive action of central acting alpha(2)-adrenoceptor agonist clonidine administered by different ways, suggesting a great sensitivity of the post-synaptic alpha(2) adrenoceptor of central nervous system. PMID- 12770511 TI - Paradoxical effects of pyruvate on cardiac contractile function under normal and high glucose in ventricular myocytes. AB - Impaired energy fuel supply and metabolism contribute to the cardiac contractile dysfunctions in diabetes. Pyruvate, a metabolic product of glycolysis and an oxidizable fuel in myocardium, has been demonstrated to enhance cardiac contractile function and alleviate hyperglycemic status. The present study was designed to examine the impact of pyruvate supplementation on cardiac mechanical function under normal glucose (NG) and high glucose (HG) conditions. Isolated adult rat ventricular myocytes were maintained in NG (5.5mM) or HG (25.5mM) medium for 24h in the absence or presence of pyruvate (5mM). Contractile indices were measured with an IonOptix edge-detection system including peak shortening (PS), maximal velocity of shortening/relengthening (+/-dL/dt), time-to-PS (TPS), time-to-90% relengthening (TR(90)), area underneath shortening (A(con)) and relengthening (A(relax)). Myocytes maintained in HG medium displayed abnormal mechanical function simulating in vivo diabetes. These abnormalities included reduced PS, +/-dL/dt, prolonged TPS/TR(90) associated with enhanced A(con) and A(relax) compared to NG myocytes. Interestingly, these HG-induced mechanical dysfunctions were completely abolished by co-incubation of pyruvate. However, NG myocytes developed mechanical defects reminiscent of those of HG following co incubation with pyruvate. These data suggest that pyruvate may paradoxical affect cardiac mechanical function under different glucose settings and therefore warrant caution when applying pyruvate therapeutically. PMID- 12770512 TI - Antinociceptive effects of Teucrium polium L total extract and essential oil in mouse writhing test. AB - In this study, the effect of Teucrium polium L, a wild-growing flowering plant belonging to the family Labiatae and found abundantly in Iran was studied on writhing test, a visceral pain model in mice. The total extract and essential oil were administered intraperitoneally 30 min before acetic acid 0.6% injection. Total extract in doses (mg x kg(-1)) of 150 (39.13%, P<0.001), 225 (65.44%, P<0.001), 300 (21.41%, P<0.01) induced reduction in writhing response as compared to control with the ED(50) 67.92 mg x kg(-1). The essential oil in doses (mg x kg(-1)) of 9.37 (35.22%, P<0.001), 18.75 (59.63%, P<0.001), 37.5 (86.60%, P<0.001), 75 (90.22%, P<0.001) and 150 (78.58%, P<0.001) induced significant reduction in writhing response when compared to control with the ED(50) of 29.41 mg x kg(-1). In order to ensure the importance of essence in production of visceral antinociception, the extract free from essential oil was prepared and injected into mice at a dose of 225 mg x kg(-1) (the most effective dose of the extract) which in comparison to total extract, the antinociception, reduced from 65.44 to 49.85% (P<0.001). It is concluded that essential oil is responsible for analgesic properties of T. polium. This study confirms the antivisceral pain properties of T. polium comparable to those of hyoscine and indomethacin and suggests a good place for it in antispasmodic therapies in human. The presence of flavonoids and sterols might be responsible for the anti-inflammatory activity of this plant. PMID- 12770513 TI - Effects of cyclooxygenase inhibitors on nitric oxide production and survival in a mice model of sepsis. AB - The effects of selective ((5,5-dimethyl-3-(3-florophenyl)-4-(4-methylsulphonyl 2(5H)-furanon); DFU) and (N-(2-cyclohexyloxy-4-nitrophenyl)-methansulphonamide; NS 398)) or non-selective (diclophenac and proquazon) inducible cyclooxygenase (COX-2) inhibitors on the survival, nitrite (stable product of nitric oxide (NO) as an index for inducible NO synthase (iNOS) activity) and 6-keto-prostaglandin F(1alpha) (6-keto-PGF(1alpha), stable product of prostacyclin as an index for COX 2 activity) production in serum, lungs, brain and/or kidney were investigated in endotoxin-induced sepsis model in mice. Endotoxin (10 mg x kg(-1), i.p.)-induced mortality was prevented by DFU, NS 398 and proquazon (0.1, 10 and 1 mg x kg(-1), respectively) and enhanced 2.6-fold with 0.1mg x kg(-1) diclophenac. Endotoxin induced increase in the serum levels of nitrite was only inhibited by 10 mg x kg( 1) diclophenac. Endotoxin caused a significant decrease only in the brain levels of nitrite without affecting 6-keto-PGF(1alpha) levels in all tissues. The decreased levels of nitrite induced by endotoxin is further reduced by 0.1mg x kg(-1) DFU and 1 and 10mg x kg(-1) diclophenac while 10 mg x kg(-1) DFU and 1mg x kg(-1) proquazon increased it. On the other hand, 1mg x kg(-1) diclophenac and proquazon, and 10 mg x kg(-1) NS 398 increased the endotoxin-induced lung levels of 6-keto-PGF(1alpha). The results suggest that the COX inhibitors may have different effects on the survival and NO production depending on tissue and dose. PMID- 12770514 TI - Effects of a glycogen synthase kinase-3 inhibitor, lithium, in adenomatous polyposis coli mutant mice. AB - Glycogen synthase kinase-3 (GSK-3) is an intermediary enzyme in various cellular pathways, and has been implicated in the pathophysiology and treatment of numerous diseases, including Alzheimer's disease, diabetes, and bipolar disorder. There is therefore in developing potent, selective GSK-3 inhibitors for the treatment of these devastating illnesses. A concern, however, is that the Wnt signaling pathway-of which GSK-3 is an important intermediary molecule-has been implicated in many human cancers. It is thus of considerable importance to determine if GSK-3 inhibitors have tumorigenic potential in systems predisposed to developing tumors by virtue of mutations of the Wnt-signaling pathway. We therefore investigated the effects of a GSK-3 inhibitor, lithium, in a murine model predisposed to the formation of tumors due to activation of the Wnt pathway the adenomatous polyposis coli (APC) multiple intestinal neoplasia (min) mouse. We found that 60 days of lithium treatment did not produce a significant increase in the number of tumors in these genetically predisposed mice. Lithium treatment resulted in a modest overall increase in the tumor size. The APC (min) mouse has previously been shown to be a robust indicator of tumorigenesis, with large increases in tumor number observed in response to a variety of agents; thus, our results suggest that lithium-and perhaps other inhibitors of GSK-3-pose a low risk for the development of cancers of the Wnt pathway. These results are consistent with the available epidemiological evidence that long-term lithium therapy does not increase cancer morbidity or mortality, but rather is associated with reduced overall mortality in bipolar disorder. PMID- 12770515 TI - Effects of Hypericum extract on the acetylcholine release: a loose patch clamp approach. AB - The St. John's Wort (Hypericum perforatum) extract (Hp) represents one of the most useful natural therapeutic agents in the treatment of moderate and mild depression. The antidepressant effects of Hp are different, by a molecular mechanism point of view, when compared to those of other antidepressant drugs and, we think, a further pharmacological characterization is needed. It is suggested that the neurochemical effects of Hp could be bind either to its activity on the uptake of some mediators in the central nervous system or to the inhibition of some enzymatic activity at the receptor level. The present study carried out with the loose patch clamp (LPC) in the mouse neuromuscular junction, indicates a potentiation of the acetylcholine (ACh) action at the mouse neuromuscular junction. The spontaneous release of ACh was unaffected by Hp indicating that neither presynaptic nor postsynaptic function are modified by Hp. Indeed, both the frequency and the amplitude of the miniature end-plate currents (mepcs) were unmodified by Hp. Furthermore, the mepcs decay time (tau), i.e. the apparent cholinergic channel life time, was significantly increased after Hp treatment. The other parameter affected was the amplitude of the evoked end-plate currents (epcs) which was constantly and in a dose dependent manner increased by Hp. These findings suggest a possible action of Hp on the acetylcholinesterase (AChE) in terms of a reduction of the degradation rate of ACh. PMID- 12770516 TI - Pharmacokinetic profile of methyldopa in the brain of sinaortic-denervated rats. AB - Pharmacokinetics of methyldopa (MD) and the effect on the dopaminergic metabolism was studied in anesthetized sham-operated (SO) and sinoaortic-denervated (SAD) rats by using the microdialysis technique. A concentric microdialysis probe was placed in the striatum or in the posterior hypothalamus. Levels of MD, homovanillic acid (HVA), and 3,4-dihydroxyphenylacetic acid (DOPAC) were measured by high pressure liquid chromatography coupled to electrochemical detection (HPLC EC). Following the i.p. administration of MD (50 mg x kg(-1), i.p.), striatal dialysates showed that this drug rapidly reaches the brain. However, in SAD rats the MD levels of dialysates were lower and decreased more rapidly compared to SO rats. On the other hand, dialysates of posterior hypothalamus showed that in SAD animals the accumulation of MD was significantly greater than in SO rats.MD does not significantly reduce the striatal production of dopaminergic metabolite DOPAC in both groups of rats. The drug also induces a decrease of DOPAC levels in hypothalamic dialysates of SO and SAD animals. On the other hand, a no significative decay of HVA levels was seen in the striatal dialysate of both groups of experimental animals. In conclusion, this study by using a microdialysis technique shows that MD has different kinetic profiles in dialysates from posterior hypothalamus and striatum of SO and SAD rats at a dose that alters dopaminergic metabolism. PMID- 12770517 TI - Changes in the blood-brain barrier permeability and in the brain tissue trace element concentrations after single and repeated pentylenetetrazole-induced seizures in rats. AB - The behaviour of brain capillary endothelium to the passage of macromolecules in single and repeated seizures conditions and its relationship to the brain trace element concentrations are the main subject of this study. For this purpose, animals were treated with either single or repeated doses of pentylenetetrazole (PTZ). As a marker of blood-brain barrier (B-BB) permeability changes, Evans Blue (EB) dye was used. Seizure activity was observed and seizure patterns and convulsion times were recorded. PTZ treatment induced generalised tonic-clonic seizure in all animals, but seizures were found to be lasting longer in single seizure group than repeated seizures group. Seizures induced by single dose PTZ treatment resulted in bilateral EB leakage in the preoptic area, caudate nucleus, putamen, thalamus, hypothalamus, midbrain, and the superior colliculus. However, repeated PTZ-induced seizures led to EB leakage in the brains of only few number of rats, and it was confined to hypothalamus, caudate nucleus, cerebellum, thalamus, and pons. On the other hand, while the levels of copper (Cu) and iron (Fe) in brain tissue were found to be decreased significantly in the repeated seizures group when compared with the other groups, the levels of zinc (Zn) did not show any differences between groups. These results indicate that the regional B-BB opening markedly differs between single and repeated PTZ-induced seizures group and this difference may be due to PTZ tolerance and changes in cerebral endothelial structure. PMID- 12770518 TI - Opioid prescription for terminally ill outpatients in a district of northern Italy: a retrospective survey. AB - A retrospective survey of the opioid prescriptions issued for cancer outpatients (2125) of the Treviso district (Veneto Region, northern Italy) during the time period 1993-2000 was carried out with the specific aims to establish the rate of opioid prescription and verify whether terminally ill outpatients (1697) who had died by the end of December 2000 received adequate opioid prescription, as compared with the Defined Daily Doses (DDDs) of opioids suggested by the World Health Organization (WHO) for a standard population. For both women and men, the maximum rate of opioid prescription was at the age of beyond 90 years. Men were more prescribed than women between 60 and 79 years of age, whereas women were more prescribed than men beyond 90 years. Opioid prescriptions concerned only morphine, buprenorphine, and pentazocine. The Anatomical Therapeutic Chemical (ATC)/DDDs analysis of opioid prescriptions indicated that total opioid use increased about 1.7-fold between 1993 and 1996, mainly because of an increase (55.4%) in morphine prescriptions. Afterwards, total opioid use remained stable, with an estimated mean annual value of 108.2+/-6.4 DDDs/million inhabitants/day. Considering terminally ill outpatients who had died by the end of December 2000, oral morphine turned out to be the most commonly prescribed opioid (64% of patients) and, among the three opioids, pentazocine was more prescribed to older patients. From the comparison between the number of "expected opioid DDDs" (i.e. days for which patients should have been prescribed opioids at the WHO recommended DDDs) and the number of prescribed opioid DDDs (i.e. days for which patients had been offered adequate opioid treatment) for individual patients, it could be estimated that only 38.1% of opioid prescriptions were adequate and a mean of 55.8 DDDs of opioids per patient were not prescribed. The opioid prescription inadequacy increased with the length of time from first prescription to patient death. In addition, a questionnaire investigation, conducted in 2001 among general practitioners of the Treviso district to evaluate their attitudes toward opioid prescribing, evidenced insufficient knowledge of general practitioners in theory and use of opioid analgesics in cancer pain management. A total of 104 (32.5%) general practitioners responded and most of them feared opioid side effects, such as respiratory depression (49.6%), constipation (41.7%), and addiction (8.7%). Furthermore, many of the respondents considered opioids capable of reducing the patient length of life (22.2%) and inappropriate to treat pediatric patients (50.6%). About 44% of the respondents experienced external pressure by relatives of patients against opioid prescription and a majority of them (58.2%) considered the recently revised Italian legislation on opioid prescription ineffective for improving their prescribing pattern. In conclusion, present data show that the vast majority of terminally ill cancer outpatients in the Treviso district received inadequate opioid prescriptions in relation to either drug daily dosage or therapy duration. Misconceptions of general practitioners of the district about opioids could contribute to the inappropriate use of these analgesics in cancer pain management. As far as we know, the ATC/DDD methodology for the opioid prescription analysis used in this survey has not been applied before. PMID- 12770520 TI - Cholinergic responses of seminal vesicles isolated from rats exposed perinatally to hydrocortisone. AB - This study was performed in order to investigate the cholinomimetic response of seminal vesicles isolated from rats treated with hydrocortisone acetate during perinatal life. At the adult phase, the body weight and the wet weight of the seminal vesicle of these animals were unchanged. However, these male rats exhibited a significant reduction in plasma testosterone concentration. A significant increase in the sensitivity of the seminal vesicle to acetylcholine was also observed. Despite this, there was a significant reduction in the maximum contractile response of the organ to this transmitter. These results indicate that exposure to hydrocortisone during the critical period of brain sexual differentiation has a long-term effect on testosterone production of male rats. In addition, physiological levels of cortisone in perinatal life are also essential to support the contractile response pattern of the seminal vesicle to acetylcholine in adult life, probably crucial to the reproductive process. PMID- 12770519 TI - Static and ELF magnetic fields enhance the in vivo anti-tumor efficacy of cis platin against lewis lung carcinoma, but not of cyclophosphamide against B16 melanotic melanoma. AB - Previous works showed that exposure to static and extremely low frequency (ELF) magnetic fields (MF) over 3 mT slows down the growth kinetics of human tumors engrafted s.c. in immunodeficient mice, reducing their metastatizing power and prolonging mouse survival. In the experiments reported here, immunocompetent mice bearing murine Lewis Lung carcinomas (LLCs) or B16 melanotic melanomas were exposed to MF and treated respectively with two commonly used anti-cancer drugs: cis-diamminedichloroplatinum (cis-platin) and N,N-bis (2-chloroethyl)tetra-hydro 2H-1,3,2-oxazaphosphorin-2-amine 2-oxide (cyclophosphamide). The experiment endpoint was survival time. The survival time of mice treated with cis-platin (3mg/kg i.p.) and exposed to MF was significantly (P<0.01) longer than that of mice treated only with cis-platin or only exposed to MF, superimposing that of mice treated with 10mg/kg i.p. of the drug, showing that MF act synergically with the pharmacological treatment. On the contrary, when mice treated with cyclophosphamide (50mg/kg i.p.) were exposed to MF no synergic effects were observed, the survival curve being exactly the same as that of mice treated with the drug alone. No clinical signs or toxicity were seen in any of the mice exposed to MF alone or along with cis-platin or cyclophosphamide treatment, compared to mice given only the two known drugs.A possible explanation for the synergic effect of MF being found in mice treated with cis-platin could be that the platinum ion stimulates radical production and that MF enhance active oxygen production bringing about changes in tumor cell membrane permeability, influencing positively the drug uptake. Alternatively, or in addition to this, it has been demonstrated that the rate of conversion of cis-platin to reactive species able to bind to DNA, is increased by localized production of free radicals by MF. PMID- 12770521 TI - Cytoprotective effects of chalcones from Zuccagnia punctata and melatonin on the gastroduodenal tract in rats. AB - The effects of 2',4'-dihydroxychalcone and 2',4'-dihydroxy-3'-methoxychalcone from Zuccagnia punctata Cav. (Fabaceae) and melatonin administration on ethanol induced gastroduodenal injury were investigated in rats. Both chalcones showed significant preventive effects in treatment with melatonin previous to the necrotising agent. These effects could be due, in part, to the radical scavenging activity of the melatonin. PMID- 12770522 TI - Extracellular glutamine to glutamate ratio may predict outcome in the injured brain: a clinical microdialysis study in children. AB - The amino acid content of hourly microdialysis samples from nine severely head injured children was examined. Of particular interest was the measurement of the excitatory amino acid glutamate, as high levels of this substance, which are associated with the excitotoxicity cascade, have been linked to high intracranial pressure and poor outcome in a similar study in adults. Interpretation of these data is complicated by many clinical and methodological factors and these are discussed in relation to the findings. Our findings from this pilot study in children confirm the associations between glutamate levels, intracranial pressure and outcome, but fail to corroborate the correlation between excitatory and structural amino acid levels seen in adult patients, that was interpreted as evidence of non-specific leakage of amino acids through damaged cell membranes. In addition, we have shown that the role of glutamine in glutamate homeostasis is an important consideration and that estimation of the extracellular glutamine/glutamate ratio may have some prognostic value in head trauma cases as there is evidence of links to clinical outcome. PMID- 12770523 TI - Effect of trichostatin a and 5'-azacytidine on transgene reactivation in U937 transduced cells. AB - In mammals, methylation of DNA within regulatory sites and histone deacetylase recruitment in transcriptional repressing domains are involved in the loss of the expression of retroviral DNA or repeat arrays transferred in cells for therapeutic purposes. Various investigation results suggest that methylation/deacetylation events are modulated by extracellular and cytoplasmic signal transduction pathways closely involved in regulating cell differentiation. To analyse gene silencing mechanisms and assess if potential pharmacological treatment affects gene silencing kinetics we transduced U937 myelomonocytic cells with a bicistronic retroviral construct carrying the herpes simplex virus thymidine kinase (HSV-TK) and beta-galactosidase (Lac-Z) genes. This vector can be employed in vivo and in vitro to render transduced cell populations susceptible to ganciclovir (GCV). We verified the effect of the histone deacetylase inhibitor Trichostatin A (TSA) alone or combined with 5'-azacytidine (5'aza-C) on transcription downmodulation. Our results indicate that in our in vitro model TSA is able to reactivate transgene expression, more efficiently and with quicker kinetics (12-24h) than 5'aza-C (36-48 h). The effect is dose dependent (between 1 and 50 nM), with no relevant toxicity. Treatment with both drugs is synergistic in gene reactivation in terms of extension and persistence, with low toxicity and no relevant differentiating effects. The cells in which transgene expression has been reactivated undergo progressive silencing, but once weekly drug treatment can maintain high transgene expression levels for more than 90 days with no evidence of selection. The results obtained by treating U937 transduced clones with TSA and/or 5'aza-C together with IL-3, G-CSF or GM-CSF cytokines suggest that transduced U937 differentiation levels do not affect basal expression, but render these cells more responsive to reactivation by TSA or TSA plus 5'aza-C, but not to 5'aza-C alone. In conclusion, the results suggest that in vitro inhibition of histone deacetylase by TSA can interfere with gene silencing mechanisms affecting 5' Moloney murine leukaemia virus long terminal repeat (MoMuLV-LTR) driven transgene expression thus providing the rationale for TSA and/or 5'aza-C administration in animal models for the translation on gene therapy applications. PMID- 12770524 TI - Troglitazone reduces heat shock protein 70 content in primary rat hepatocytes by a ubiquitin proteasome independent mechanism. AB - Troglitazone (TRG) is an antidiabetic agent that increases the insulin sensitivity of target tissues in non-insulin-dependent diabetes mellitus. Therapy with troglitazone has been associated with severe hepatic injury in a small percentage of patients and the mechanism of TRG-induced hepatotoxicity remains unclear. A family of highly conserved stress proteins identified as heat shock proteins (Hsps), are well-known to protect cells against a wide variety of toxic conditions such as extreme temperature changes, oxidative stress and toxic drugs. The stress-inducible Hsp 70 protein is one of the best-known endogenous factors protecting cells from injury under various stress conditions. Here we examined the effects of TRG on Hsp 70 mRNA and protein expression in primary cultures of rat hepatocytes. We also investigated the effects of TRG in an in vivo model by examining Hsp 70 protein levels in livers prepared from C57 mice fed a 0.2% dietary admixture of TRG. Levels of Hsp 70 mRNA increased in a concentration dependent manner in rat hepatocytes treated for 8h with increasing concentrations of TRG. However, Hsp 70 protein levels decreased significantly in cells treated with increasing concentrations of TRG. C57 mice fed a 0.2% admixture of TRG for 10 days, also demonstrated decreased liver Hsp 70 protein levels. To investigate whether TRG decreased Hsp 70 protein levels by activating the ubiquitin proteasome pathway, cells were pretreated with 10 microM lactacystin, a potent and specific inhibitor of this pathway. Lactacystin pretreatment failed to prevent TRG-induced decrease in Hsp 70 protein. The data suggests that TRG induced effects may be mediated through another system of regulated proteolysis or may involve a post-transcriptional regulator mechanism. The mechanism of TRG induced hepatotoxicity remains unclear, however, the effects induced by TRG on Hsp 70 may, in part, play a role. PMID- 12770525 TI - Risk taking and coping strategies among Israeli adolescents. AB - The research sample consisted of 140 Israeli 10th- and 11th-graders who completed questionnaires about risk-taking behaviours and coping strategies. The study assessed the associations between risk behaviours and perceptions of benefit and risk in these behaviours as measured by a questionnaire based on the Risk Involvement and Perception Scale (Siegel, A. W., Cousins, J. H., Rubovits, P., Parsons, J. T., Lavery, B., Crowley, C. L. (1994). Journal of Emotional and Behavioral Disorders, 2, 89-98); and the associations between risk measures and problem-focused, support and avoidance coping strategies as assessed by the COPE scale (Carver, C. S., Scheier, M., Weintraub, J. K. (1989). Journal of Personality and Social Psychology, 56, 267-283). The data were analysed using correlations and multiple regressions. The main results showed that high involvement in risk behaviour was positively related to the perception of benefit in risk behaviour, but perception of risk was not related to involvement in risk behaviour. Most significantly, the use of avoidance coping strategies was related to high involvement in risk behaviours. The results suggest that perception of benefit and the use of avoidance coping strategies may constitute significant factors in the decision to take risks, with implications for intervention programs aimed at adolescents. PMID- 12770526 TI - The experience of loneliness of Canadian and Czech youth. AB - The present study examined the influence of cultural background on adolescents' experience of loneliness. Eighty Canadians and 168 adolescents from the Czech Republic formed the participants pool. They answered a 30-item yes/no questionnaire. The questionnaire is composed of five subscales, namely: Emotional distress, Social inadequacy and alienation, Growth and discovery, Interpersonal isolation, and Self-alienation. Results revealed a mixed pattern of differences between the two cultures, with Canadian youth scoring higher than the Czechs only on Emotional distress, while that trend was reversed in Social inadequacy and alienation and Interpersonal isolation. Gender differences within cultures were also examined. PMID- 12770527 TI - Urgent adolescent psychiatric consultation: from the accident and emergency department to inpatient adolescent psychiatry. AB - The Rapid Response Model (RRM) provides psychiatric services to children and adolescents seen at the Accident and Emergency (A&E) department or at the Urgent Consultation Clinic of the Child and Adolescent Psychiatry Division the next day. In a naturally occurring experiment, the RRM was introduced, withdrawn and restarted. When RRM was withdrawn at one site, it was implemented at another. The RRM reduced nighttime Emergency Consultations and inpatient admissions from A&E, while it increased daytime consultations and daytime admissions. The RRM provided timely, organized emergency psychiatric services. A&E staff expressed satisfaction with the service. PMID- 12770528 TI - Leaving high school: the influence and consequences for psychological well-being and career-related confidence. AB - This paper examines the well-being and career decision-making self-efficacy (CDMSE) of adolescents before and after leaving school, and tests for the changes in these variables as a result of leaving school. While at high school, 309 students were assessed on levels of school achievement, well-being (psychological distress, self-esteem, life satisfaction) and CDMSE. Nine months after leaving school, 168 of these students completed the above surveys and measures of their access to the latent (e.g. social contact, time structure) and manifest (i.e. financial) benefits of employment, and work commitment. At T2, 21% were full-time students, 35% were full-time students who were also working part-time, 22% were employed in full-time jobs, and 21% were in the labour market but not employed full-time. These groupings were differentiated at T2 on aspects of well-being, self-efficacy, and access to the latent and manifest benefits of work, and at T1 on aspects of well-being and confidence. Leaving school improved well-being and confidence for some. One group was disadvantaged by having poorer well-being while at school, which predisposed them to disadvantage in the labour market. Results are discussed in relation to models of well-being and drift/social causation. PMID- 12770529 TI - The social determinants of youth gambling in South Australian adolescents. AB - Recent Australian studies (Moore, S.M., and Ohtsuka, K. (1997). Journal of Gambling Studies, 13, 207-236) have revealed a strong youth interest in gambling in Australia, as reflected in current participation levels, future intentions and attitudes. Implicit in much of this attitudinal research is that youth gambling is strongly influenced by the familial, social and cultural norms to which young people are exposed. In this paper, we investigate the hypothesis that gambling can be understood in terms of variations in economic socialization, namely, the way in which children learn about money, risk, and saving. A school survey of 505 adolescents (aged 15-17 years) showed that over 60% of adolescents were gambling annually and that 3.5% scored in the problematic range on the DSM-IV-J (Fisher, S.E. (1999). Addiction Research, 7, 509-538). More frequent gambling was associated with parental and peer gambling and pro-gambling attitudes, but unrelated to adolescents' attitudes towards economic concepts. Nevertheless, in partial support of the hypotheses, adolescents whose parents taught them about keeping to a budget, saving money, and maintaining their finances were less likely to express an interest in future gambling. PMID- 12770530 TI - Drug-use initiation and conduct disorder among adolescents in drug treatment. AB - This study investigated effects of drug-use initiation and conduct disorder (CD) among 1031 adolescents who participated in the Drug Abuse Treatment Outcomes Studies for Adolescents (DATOS-A) sponsored by the National Institute on Drug Abuse (NIDA). The mean age of first drug use was 12.7 (S.D.=2.2), 57% met DSM-III R criteria for CD, and earlier initiators were more likely to have CD. About 78% of the adolescents with CD reported that their first CD symptom occurred prior to drug-use initiation. The proportions of adolescents who had prior treatment were similar (about 28%) across all groups, but earlier initiators reported a greater number of treatment episodes and younger ages at their first treatment. Conduct disordered adolescents revealed greater problems prior to DATOS-A treatment, but they appeared to be more motivated and ready for treatment. Although adolescents with CD still showed worse outcomes after treatment, the impact of CD appeared to lessen when pretreatment differences were controlled. To a lesser extent, adolescents who began using drugs at earlier ages had greater alcohol and drug use and other problems at intake, but their treatment outcomes appeared to be similar to later initiators. There were few significant interaction effects of initiation and CD. Findings from this study highlight the importance of better understanding the progression of drug use, treatment utilization, and psychiatric comorbidity among adolescents with substance abuse problems. PMID- 12770531 TI - Individual differences in attachment-autonomy configurations: linkages with substance use and youth competencies. AB - Variations in attachment-autonomy configurations are explored as these relate to substance use and several adolescent competencies. Questionnaires completed by 470 university students included measures of parental attachment, autonomy (both emotional autonomy and self-reliance), substance use, problems associated with substance use, social competencies, and coping. Analyses of subgroups representing four attachment-autonomy patterns, derived from cluster analysis, showed higher competency levels and fewer problems related to substance use for the group combining strong attachment and self-reliance coupled with low levels of emotional autonomy. A subgroup characterized by low father but moderate mother attachment security differed in a number of ways from a subgroup reporting low attachment to both parents. Findings support the utility of a typological approach in investigations of adolescent attachment and autonomy. PMID- 12770532 TI - Minimizing respondent attrition in longitudinal research: practical implications from a cohort study of adolescent drinking. AB - The methods used to maximize retention in a longitudinal study of adolescent drinking are discussed. Data were collected at three time points: at recruitment to the study, after nine months and at 18 months. Strategies to minimize attrition included the collection of detailed contact information, incentives for participation, postcard and telephone reminders and telephone interviews. Ninety six percent of the original sample completed the first follow-up questionnaire, 92% completed the second and the study lost contact with just 3% of participants. The success of the current project is notable as this type of population is notoriously difficult to retain in longitudinal studies. PMID- 12770533 TI - Young people's social representations of a sexual experience. PMID- 12770537 TI - Bovine viral diarrhoea virus antigenic diversity: impact on disease and vaccination programmes. AB - Bovine viral diarrhoea virus (BVDV) infections in cattle are associated with a variety or "diverse" clinical forms. These include digestive tract, respiratory, foetal (varied, dependent on foetal age), haemorrhagic and systemic diseases such as mucosal disease, and immunosuppression and inapparent infections. The BVDV isolates themselves are "diverse" with genotype differences based on nucleotide sequences, antigenic variability and biotypes (presence or absence of cytopathology in cell culture). Two predominant genotypes are present in the US, BVDV1 and BVDV2. There are subtypes of BVDV1, namely BVDV1a and BVDV1b. Examination of BVDV isolates from cattle derived from diagnostic laboratory submissions indicates that BVDV1b subtype isolates were as prevalent if not more prevalent than BVDV1a isolates. There was an almost equal distribution of BVDV1b and BVDV1a isolates from cattle with history of respiratory disease, and more isolates, 6 versus 2, of BVDV1b than for BVDV1a in necropsy cases of pneumonia. There were significant antibody titre differences in sera from calves receiving modified live virus vaccines containing BVDV1a, with the BVDV1b antibody titres being significantly lower. A survey of the US licensed and marketed BVDV vaccines indicates that only one vaccine contains BVDV1b with the others containing BVDV1a or undesignated BVDV1. PMID- 12770538 TI - Gene expression changes in BVDV2-infected MDBK cells. AB - Bovine viral diarrhoea virus (BVDV) is a ubiquitous viral pathogen of cattle. The virus exists as one of two biotypes, cytopathic and non-cytopathic, based on the ability to induce cytopathic effect in cell culture. The non-cytopathic biotypes are able to establish non-apparent, persistent infections in both cell culture and in bovine foetuses of fewer than 150 days gestation. The mechanism by which viral tolerance is established is unknown. To examine the changes in gene expression that occur following infection of host cells with BVDV, serial analysis of gene expression (SAGE), a global gene expression technology was used. SAGE, a sequence-based technology, allows quantification of virtually every transcript in a cell type without prior sequence information. Transcript expression levels and identities are determined by DNA sequencing of libraries composed of 14 base DNA fragments (tags) derived from the 3' end of each cellular mRNA transcript. Comparison of data obtained from non-infected and BVDV2-infected cell libraries revealed a number of changes in gene expression. Many of these transcriptional changes could be placed into distinct biochemical pathways or functions. Both alpha and beta tubulins were downregulated, indicating possible dysfunction in cell division and other functions where microtubules play a major role. Expression of several genes encoding proteins involved in energy metabolism were downregulated, indicating possible decreased ATP synthesis. Genes encoding proteins involved in protein translation and post-translational modifications were generally upregulated. These data indicate that following infection with BVDV, changes in gene expression occur that are beneficial for virus replication while placing the cell at a metabolic disadvantage. PMID- 12770539 TI - A novel simple one-step single-tube RT-duplex PCR method with an internal control for detection of bovine viral diarrhoea virus in bulk milk, blood, and follicular fluid samples. AB - A simple one-step single-tube RT-PCR method was developed for detection of bovine viral diarrhea virus (BVDV) in bulk milk, blood and follicular fluid samples. A set of universal primers (UTR DL1/4) was designed for RT-PCR to detect BVDV type I and II viruses simultaneously and was tested for efficacy in comparison to published primers for two type strains, 42 field isolates, and 95 diagnostic samples. The sensitivity (100%) and specificity (96.2%) of the RT-PCR assay, with the universal primers for 95 diagnostic samples, were equal to those of virus isolation. An internal control targeting a bovine actin gene was also included in the same reaction tube to monitor RNA preparation and RT-PCR procedure. A total of 632 specimens (378 bulk milk, 140 blood, and 112 follicular samples) were tested in the year 2000 by the RT-duplex PCR assay in parallel with virus isolation. The one-step single-tube RT-duplex PCR with the universal primers UTR DL1/4 was sensitive, specific, less complicated and cost effective for detection of BVDV in various types of specimens. PMID- 12770540 TI - BVDV and innate immunity. AB - Infection with bovine viral diarrhea virus (BVDV) is prevalent in the cattle population worldwide. The virus exists in two biotypes, cytopathic and non cytopathic, depending on the effect of the viruses on cultured cells. BVDV may cause transient and persistent infections which differ fundamentally in the host's antiviral immune response. Transient infection may be due to both cytopathic and non-cytopathic biotypes of BVDV and leads to a specific immune response. In contrast, only non-cytopathic BVD viruses can establish persistent infection as a result of infection of the embryo early in its development. Persistent infection is characterized by immunotolerance specific for the infecting viral strain. In this paper we discuss the role of innate immune responses in the two types of infection. In general, both transient and persistent infections are associated with an increased frequency of secondary infections. Associated with the increased risk of such infections are, among others, impaired bacteria killing and decreased chemotaxis. Interestingly, non cytopathic BVDV fails to induce interferon type I in cultured bovine macrophages whereas cytopathic biotypes readily trigger this response. Cells infected with non-cytopathic BVDV are also resistant to induction of interferon by double stranded RNA, a potent interferon inducer signalling the presence of viral replication in the cell. Thus, non-cytopathic BVDV may dispose of a mechanism suppressing a key element of the antiviral defence of the innate immune system. Since interferon is also important in the activation of the adaptive immune response, suppression of this signal may be essential for the establishment of persistent infection and immunotolerance. PMID- 12770541 TI - Bovine viral diarrhoea eradication and control programmes in Europe. AB - The economic impact of BVDV infections has led a number of countries in Europe to start eradication or control programmes. While in both cases the primary step is identification and elimination of persistently infected (PI) animals, the strategy applied thereafter is dependent on the density and seroprevalence of the regional cattle population. One of the first countries to design and implement an eradication programme was Sweden in 1993, a country with a relatively low cattle density and no vaccination. For screening, an indirect antibody ELISA for serum, milk and bulk milk samples is being used. The basics of the Swedish model are no vaccination, voluntary participation, and financing of the entire scheme by the subscribing farmers. BVDV-free herds are certified and permanently checked. While in 1993 only about 35% of the herds were seronegative, about 87% were BVDV-free in 2001. The aim of control programmes in high density areas with high seroprevalence is to minimize economic losses by reducing the incidence of PI animals and thereby virus circulation (German model). Participation is voluntary, and parts of the costs are carried by the public animal insurance (Tierseuchenkasse). Screening is performed using an antigen capture ELISA with blood or serum. In Lower Saxony, for example, a herd is declared BVDV unsuspicious if all animals up to 36 months are BVDV antigen negative and the female offspring older than six months is vaccinated twice (an inactivated vaccine is used for basic immunization, and an attenuated live virus vaccine for boosting). PMID- 12770542 TI - Lesions and tissue distribution of viral antigen in severe acute versus subclinical acute infection with BVDV2. AB - Differences in the distribution and spread of viral antigen, development of lesions and correlation between presence of viral antigen and lesions were compared between a low and highly virulent strain of BVDV2. Two groups of two week- to two-month-old colostrum-deprived calves were inoculated intranasally with the naturally occurring low virulent BVDV2 strain 28508-5 or the highly virulent strain 1373. To study the sequence of virus spread and lesion development, calves were necropsied at days three, six, eight-nine and 12 to 14 post inoculation (pi). Viral antigen was detected by the indirect immunoperoxidase method in cryostat sections and lesions were evaluated in H&E stained paraffin sections. Clinical signs and changes in lymphocyte and thrombocyte numbers confirmed the difference in virulence between the two strains. Both strains showed comparable initial infection and spread at day three pi. At day six pi, they were found widespread in lymphoid tissues and multifocally in intestinal mucosa. Lesions were very mild despite the large amount of antigen in the lymphoid tissues. After day six pi, differences between the low and highly virulent strains became more prominent. The strain of low virulence was cleared from the tissues, but there was a transient phase of depletion. The highly virulent strain continued to spread to different organs and there was severe depletion of lymphoid tissues without recovery. PMID- 12770543 TI - Maternal antibody blocks humoral but not T cell responses to BVDV. AB - Bovine viral diarrhoea virus (BVDV) contributes significantly to health-related economic losses in the beef and dairy industry. Antibodies of maternal origin can be protective against BVDV infection, however, calves with low titres of maternal antibody or that do not receive colostrum may be at risk for acute BVDV infection. Interference by high titres of maternal antibodies prevents the development of an antibody response following vaccination with either a killed or attenuated BVDV vaccine. However, the T cell mediated immune response to BVDV may be generated in the absence of a detectable serum neutralizing antibody response. Two trials were conducted to evaluate the potential to elicit T cell mediated immune responses to BVDV in calves with circulating maternal antibody to BVDV. In the first trial, calves with high levels of circulating maternal antibody to BVDV 1 and BVDV 2 were experimentally infected with BVDV 2 (strain 1373) at two to five weeks of age. The T-cell mediated immune responses of the experimentally infected calves and non-infected calves were monitored monthly until circulating maternal antibody was no longer detectable in either treatment group. Calves experimentally infected with BVDV developed BVDV specific CD4(+), CD8(+), and delta T cell responses while high levels of maternal antibody were circulating. A second challenge with BVDV 2 (strain 1373) was performed in the experimentally infected and control calves once maternal antibody could no longer be detected. Previous exposure to BVDV in the presence of maternal antibody protected calves from clinical signs of acute BVDV infection compared to the control calves. In the second trial, three groups of calves with circulating maternal antibody to BVDV were given either a modified live vaccine (MLV) containing BVDV 1 and BVDV 2, a killed vaccine containing BVDV 1 and BVDV 2, or no vaccine, at seven weeks of age. Serum neutralizing antibody levels and antigen specific T cell responses were monitored for 14 weeks following vaccination. Calves vaccinated with MLV BVDV developed BVDV 1 and BVDV 2 specific CD4(+)T cell responses, and BVDV 2 specific gammadelta T cell responses, in the presence of maternal antibody. Vaccination with killed BVDV did not result in the generation of measurable antigen specific T cell immune responses. In this trial, a second vaccination was performed at 14 weeks to determine whether an anamnestic antibody response could be generated when calves were vaccinated in the presence of maternal antibody. Calves vaccinated with either a MLV or killed BVDV vaccine while they had maternal antibody developed an anamnestic antibody response to BVDV 2 upon subsequent vaccination. The results of these trials indicate that vaccinating young calves against BVD while maternal antibody is present may generate BVDV specific memory T and B cells. The data also demonstrated that seronegative calves with memory T and B cells specific for BVDV may be immune to challenge with virulent BVDV. PMID- 12770544 TI - BVDV genotypes and biotypes: practical implications for diagnosis and control. PMID- 12770545 TI - The persistence of bovine viral diarrhea virus. AB - Bovine viral diarrhoea virus (BVDV) has a unique capacity to cause persistent infections of foetuses exposed within the first 150 days of gestation. Preventing foetal BVDV infection will aid in improved control. This unique ability gives BVDV a selective advantage allowing continual mutation and antigenic variation within cattle populations. Therefore, BVDV has become widespread and causes economic losses due to respiratory, reproductive and enteric disease. Vaccination (modified-live or killed) can provide some protection from acute disease and the development of persistently infected foetuses. However, vaccination programmes alone cannot control or eliminate BVDV. In naturally exposed and vaccinated herds, BVDV infections are not self-limiting and may persistent over time. This underscores the ability of the BVDV genome to remain fluid and adapt under selective pressures. Factors influencing persistence of BVDV infections in cattle populations include: non-lytic infections; evasion of host immune responses; foetal infections; acute infections; management practices; contaminated biologics; secondary hosts; defective replicated intermediates; antigenic variation; and replication in privileged anatomical sites. PMID- 12770546 TI - Economic impact of BVDV infection in dairies. AB - The detrimental effects of bovine virus diarrhoea virus (BVDV) infections include reduced milk production, reduced reproductive performance, growth retardation, increased occurrence of other diseases, unthriftiness, early culling and increased mortality among young stock. These losses have been documented in several case descriptions and to some extent quantified in epidemiological studies. The detrimental effects together with information on population structure, incidence of infection and monetary value of production losses have been included in different models for estimating economic losses and benefits of different control strategies. This paper reviews different studies and methods for estimating economic losses and the economic effect of control strategies on both the local herd level and the national herd level. The estimated losses in individual herd outbreaks have varied from a few thousand up to $100000. There seems to be no universal truth for determining most optimal strategy at the herd level as it depends on herd-specific conditions. Most estimations of the losses at the national level range between 10 and 40 million $ per million calvings. In the few countries that have introduced eradication campaigns, the programmes have been shown to be cost effective. However, selection of a control strategy should always rely on thorough epidemiological investigations conducted under the same conditions in which the programme is going to be applied. PMID- 12770547 TI - Trends in the BVDV serological response in the Upper Midwest. AB - Bovine viral diarrhoea continues to be an important disease affecting both beef and dairy animals of all ages. One of the quickest means of measuring bovine viral diarrhoea virus (BVDV) exposure and infection in the herd is a serum neutralization (SN) assay. Type 1 and type 2 BVDV SN results from the Animal Disease Research and Diagnostic Laboratory at South Dakota State University were collected over a seven-year period (1995-2001) to determine any trends. These results indicated that in 1996, 31% of the animals had titres > or =512 while in 2001, 74% of the titres were > or = 512. There has been a progressive increase over the seven-year period in the number of cases with titres > or = 512 with the exception of 1999 when there was a slight decrease. When analyzing the titres greater than 512, a further increase was seen. In 1995, 80% of the titres were < or = 1024 and 20% were 2048 and no titres were >2048. In 2001, 47% of the antibody levels were < or = 1024 while 53% were < or =2048 and 30% were >4096. The most dramatic increase in titres occurred in 1997 and the percentage of animals with titres from 512-8192 has remained fairly constant for the last five years (1997-2001). This increase is in part due to more extensive use of vaccination but probably also reflects a rise in field infections. In the future, standardizing existing BVDV SN serology along with developing new BVDV serology methods is necessary to provide continuity for any full-scale eradication programme. PMID- 12770548 TI - The amygdala is part of the behavioural reinforcement system modulating long-term potentiation in rat hippocampus. AB - Long-term potentiation (LTP) in the dentate gyrus can be modulated and prolonged by emotional/motivational influences when concurrently activated. A similar effect on LTP can be obtained by stimulating the amygdala, suggesting that this limbic structure might be part of the neural system involved in behavioural reinforcement. To confirm this we have performed a series of experiments in which the basolateral amygdala was either temporary inactivated by injection of lidocaine or permanently lesioned electrolytically. Both manipulations completely blocked the reinforcing effect of a motivational stimulus (drinking after 24-h deprivation) on LTP at the perforant pathway-dentate gyrus synapses, whilst leaving intact the non-reinforced potentiation. These results demonstrate that the basolateral amygdala is a key structure within the system involved in the modulatory interaction between the affective status of the animal and the mechanisms of functional plasticity. PMID- 12770550 TI - Calcitonin gene-related peptides: their binding sites and receptor accessory proteins in adult mammalian skeletal muscles. AB - This work addresses the presence, pharmacological properties, and anatomical localization of calcitonin gene-related peptide-alpha (CGRPalpha) binding sites and the receptor's accessory proteins in endplate-enriched and non-endplate muscle membrane samples from adult rat gracilis muscles. We examined the binding of (125)I-[Tyr(0)]-CGRPalpha, the competitive binding of CGRPalpha analogs, the immunohistochemical localization of the receptor's accessory proteins, and Western blots of the receptor component protein. Results show that: (a). (125)I [Tyr(0)]-CGRPalpha binding is saturable, specific, and consistent with the presence of a homogeneous population of binding sites (Hill coefficients=1.0) in endplate and non-endplate samples exhibiting dissociation constants of 0.39 nM and 0.38 nM, respectively; (b). the density of binding sites in the endplate samples (71.0 fmoles/mg protein) is considerably higher than that in their non endplate counterparts (34.6 fmoles/mg protein); (c). unlabeled CGRPalpha, hCGRP8 37 and calcitonin compete with the radioligand with the same order of potency in the endplate and non-endplate samples; and (d). the localization of the receptor accessory proteins, including the receptor activity-modifying protein (RAMP1) and the receptor component protein (RCP), for the most part matches that of the motor end-plates. Thus, gracilis muscles express CGRPalpha-specific binding sites which are predominantly localized in the muscle's motor endplate regions where RAMP1, RCP, CGRPalpha, acetylcholine receptors, and acetylcholinesterase are detected in high concentrations. These findings imply that the CGRPalpha binding sites reflect the presence of physiologically functional receptors with a pharmacological profile consistent with that of the CGRPalpha receptor type 1 (CGRP1). When considered together with earlier studies on the same neuromuscular preparation, the present work further suggests that the motoneuron-dependent trophic control of acetylcholine receptors and acetylcholinesterase in skeletal muscle endplates is partly mediated by nerve-derived CGRPalpha activating specific receptors which are highly sensitive to the truncated peptide hCGRP8-37. PMID- 12770549 TI - Molecular and functional changes in voltage-dependent Na(+) channels following pilocarpine-induced status epilepticus in rat dentate granule cells. AB - Status epilepticus (S.E.) is known to lead to a large number of changes in the expression of voltage-dependent ion channels and neurotransmitter receptors. In the present study, we examined whether an episode of S.E. induced by pilocarpine in vivo alters functional properties and expression of voltage-gated Na(+) channels in dentate granule cells (DGCs) of the rat hippocampus. Using patch clamp recordings in isolated DGCs, we show that the voltage-dependent inactivation curve is significantly shifted toward depolarizing potentials following S.E. (half-maximal inactivation at -43.2+/-0.6 mV) when compared with control rats (-48.2+/-0.8 mV, P<0.0001). The voltage-dependent activation curve is significantly shifted to more negative potentials following S.E., with half maximal activation at -28.6+/-0.8 mV compared with -25.8+/-0.9 mV in control animals (P<0.05). The changes in voltage dependence resulted in an augmented window current due to increased overlap between the activation and inactivation curve. In contrast to Na(+) channel voltage-dependence, S.E. caused no changes in the kinetics of fast or slow recovery from inactivation. The functional changes were accompanied by altered expression of Na(+) channel subunits measured by real time reverse transcription-polymerase chain reaction in dentate gyrus microslices. We investigated expression of the pore-forming alpha subunits Na(v)1.1-Na(v)1.3 and Na(v)1.5-Na(v)1.6, in addition to the accessory subunits beta(1) and beta(2). The Na(v)1.2 and Na(v)1.6 subunit as well as the beta(1) subunit were persistently down-regulated up to 30 days following S.E. The beta(2) subunit was transiently down-regulated on the first and third day following S.E. These results indicate that differential changes in Na(+) channel subunit expression occur in concert with functional changes. Because coexpression of beta subunits is known to robustly shift the voltage dependence of inactivation in a hyperpolarizing direction, we speculate that a down-regulation of beta-subunit expression may contribute to the depolarizing shift in the inactivation curve following S.E. PMID- 12770551 TI - Large variability in synaptic N-methyl-D-aspartate receptor density on interneurons and a comparison with pyramidal-cell spines in the rat hippocampus. AB - Pyramidal cells receive input from several types of GABA-releasing interneurons and innervate them reciprocally. Glutamatergic activation of interneurons involves both alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) and N-methyl-D-aspartate (NMDA) type glutamate receptors expressed in type I synapses, mostly on their dendritic shafts. On average, the synaptic AMPA receptor content is several times higher on interneurons than in the spines of pyramidal cells. To compare the NMDA receptor content of synapses, we used a quantitative postembedding immunogold technique on serial electron microscopic sections, and analysed the synapses on interneuron dendrites and pyramidal cell spines in the CA1 area. Because all NMDA receptors contain the obligatory NR1 subunit, receptor localisation was carried out using antibodies recognising all splice variants of the NR1 subunit. Four populations of synapse were examined: i). on spines of pyramidal cells in stratum (str.) radiatum and str. oriens; ii). on parvalbumin-positive interneuronal dendritic shafts in str. radiatum; iii). on randomly found dendritic shafts in str. oriens and iv). on somatostatin-positive interneuronal dendritic shafts and somata in str. oriens. On average, the size of the synapses on spines was about half of those on interneurons. The four populations of synapse significantly differed in labelling for the NR1 subunit. The median density of NR1 subunit labelling was highest on pyramidal cell spines. It was lowest in the synapses on parvalbumin-positive dendrites in str. radiatum, where more than half of these synapses were immunonegative. In str. oriens, synapses on interneurons had a high variability of receptor content; some dendrites were similar to those in str. radiatum, including the proximal synapses of somatostatin-positive cells, whereas others had immunoreactivity for the NR1 subunit similar to or higher than synapses on pyramidal cell spines. These results show that synaptic NMDA receptor density differs between pyramidal cells and interneurons. Some interneurons may have a high NMDA receptor content, whereas others, like some parvalbumin-expressing cells, a particularly low synaptic NMDA receptor content. Consequently, fast glutamatergic activation of interneurons is expected to show cell type-specific time course and state dependent dynamics. PMID- 12770552 TI - A saturated-fat diet aggravates the outcome of traumatic brain injury on hippocampal plasticity and cognitive function by reducing brain-derived neurotrophic factor. AB - We have conducted studies to determine the potential of dietary factors to affect the capacity of the brain to compensate for insult. Rats were fed with a high-fat sucrose (HFS) diet, a popularly consumed diet in industrialized western societies, for 4 weeks before a mild fluid percussion injury (FPI) or sham surgery was performed. FPI impaired spatial learning capacity in the Morris water maze, and these effects were aggravated by previous exposure of the rats to the action of the HFS diet. Learning performance decreased according to levels of brain-derived neurotrophic factor (BDNF) in individual rats, such that rats with the worst learning efficacy showed the lowest levels of BDNF in the hippocampus. BDNF immunohistochemistry localized the decreases in BDNF to the CA3 and dentate gyrus of the hippocampal formation. BDNF has a strong effect on synaptic plasticity via the action of synapsin I and cAMP-response element-binding protein (CREB), therefore, we assessed changes in synapsin I and CREB in conjunction with BDNF. Levels of synapsin I and CREB decreased in relation to decreases in BDNF levels. The combination of FPI and the HFS diet had more dramatic effects on the active state (phosphorylated) of synapsin I and CREB. There were no signs of neurodegeneration in the hippocampus of any rat group assessed with Fluoro-Jade B staining. The results suggest that FPI and diet impose a risk factor to the molecular machinery in charge of maintaining neuronal function under homeostatic and challenging situations. PMID- 12770553 TI - Expression of a functional Fas death receptor by human foetal motoneurons. AB - The expression of the apoptosis inducer Fas (CD95/APO-1) surface receptor by human foetal neurons was investigated in vitro and ex vivo. Immunofluorescence studies of brain and spinal cord cells in primary cultures and of cryosections obtained from 9- and 10-week-old human foetuses, respectively, showed that all Fas-expressing cells were motoneurons (5.3 and 4.2% of the neurons in brain or spinal cord cultures, respectively) on the basis of morphology, reactivity with the monoclonal antibody SMI-32, a mostly motoneuronal marker and acetylcholine esterase expression. Fas was undetectable on the other cell types in culture. The ability of Fas to induce apoptosis of cultured cells from both tissues was determined by using the terminal transferase (TdT)-mediated dUTP nick-end labelling (TUNEL) method combined with the same double-staining procedure. Under basal culture conditions, about 9% of cells, all glial fibrillary acidic protein expressing astrocytes, were apoptotic. After a 48-h incubation with Fas ligand, mean 28.5% of brain motoneurons and 29.4% of spinal motoneurons underwent apoptosis, with an inhibition by Z-IETD-FMK, a caspase-8 inhibitor. Hence, Fas appears to be functional through a caspase-8-dependent pathway in a subpopulation of human foetal motoneurons. PMID- 12770554 TI - C-Jun N-terminal kinases/c-Jun and p38 pathways cooperate in ceramide-induced neuronal apoptosis. AB - Understanding the regulation of the apoptotic program in neurons by intracellular pathways is currently a subject of great interest. Recent results suggest that c Jun N-terminal kinases (JNK), mitogen-activated protein kinases and the transcription factor c-Jun are important regulators of this cell death program in post-mitotic neurons following survival-factor withdrawal. Our study demonstrates that ceramide levels increase upon survival-factor withdrawal in primary cultured cortical neurons. Furthermore, survival-factor withdrawal or addition of exogenous c(2)-ceramide induces JNK pathway activation in these cells. Western blot analyses of JNK and c-Jun using phospho-specific antibodies reveal that JNK and subsequent c-Jun phosphorylation occur hours before the initiation of apoptosis, reflected morphologically by neurite retraction and fragmentation, cell-body shrinkage and chromatin fragmentation. Immunocytochemistry using the same antibodies shows that phospho-JNK are localized in the neurites of control neurons and translocate to the nucleus where phospho-c-Jun concurrently appears upon ceramide-induced apoptosis. To determine if ceramide-induced c-Jun activation is responsible for the induction of the apoptotic program, we performed transient transfections of a dominant negative form of c-Jun, truncated in its transactivation region. Our results show that DNc-Jun partially protects cortical neurons from ceramide-induced apoptosis. Treatment of dominant negative c-Jun-expressing neurons with the pharmacological inhibitor of p38 kinase, SB203580, completely blocked neuronal death. Thus our data show that p38 and JNK/c-Jun pathways cooperate to induce neuronal apoptosis. PMID- 12770556 TI - Sox1-deficient mice suffer from epilepsy associated with abnormal ventral forebrain development and olfactory cortex hyperexcitability. AB - Mutations in several classes of embryonically-expressed transcription factor genes are associated with behavioral disorders and epilepsies. However, there is little known about how such genetic and neurodevelopmental defects lead to brain dysfunction. Here we present the characterization of an epilepsy syndrome caused by the absence of the transcription factor SOX1 in mice. In vivo electroencephalographic recordings from SOX1 mutants established a correlation between behavioral changes and cortical output that was consistent with a seizure origin in the limbic forebrain. In vitro intracellular recordings from three major forebrain regions, neocortex, hippocampus and olfactory (piriform) cortex (OC) showed that only the OC exhibits abnormal enhanced synaptic excitability and spontaneous epileptiform discharges. Furthermore, the hyperexcitability of the OC neurons was present in mutants prior to the onset of seizures but was completely absent from both the hippocampus and neocortex of the same animals. The local inhibitory GABAergic neurotransmission remained normal in the OC of SOX1 deficient brains, but there was a severe developmental deficit of OC postsynaptic target neurons, mainly GABAergic projection neurons within the olfactory tubercle and the nucleus accumbens shell. Our data show that SOX1 is essential for ventral telencephalic development and suggest that the neurodevelopmental defect disrupts local neuronal circuits leading to epilepsy in the SOX1-deficient mice. PMID- 12770555 TI - Immunocytochemical evidence that amyloid beta (1-42) impairs endogenous antioxidant systems in vivo. AB - Amyloid beta, the major constituent of the senile plaques in the brains of patients with Alzheimer's disease, is cytotoxic to neurons and has a central role in the pathogenesis of the disease. We have previously demonstrated that potent antioxidants idebenone and alpha-tocopherol prevent learning and memory impairment in rats which received a continuous intracerebroventricular infusion of amyloid beta, suggesting a role for oxidative stress in amyloid beta-induced learning and memory impairment. To test the hypothesis, in the present study, we investigated alterations in the immunoreactivity of endogenous antioxidant systems such as mitochondrial Mn-superoxide dismutase, glutathione, glutathione peroxidase and glutathione-S-transferase following the continuous intracerebroventricular infusion of amyloid beta for 2 weeks. The infusion of amyloid beta (1-42) resulted in a significant reduction of the immunoreactivity of these antioxidant substances in such brain areas as the hippocampus, parietal cortex, piriform cortex, substantia nigra and thalamus although the same treatment with amyloid beta (40-1) had little effect. The alterations induced by amyloid beta (1-42) were not uniform, but rather specific for each immunoreactive substance in a brain region-dependent manner. These results demonstrate a cytological effect of oxidative stress induced by amyloid beta (1-42) infusion. Furthermore, our findings may indicate a heterogeneous susceptibility to the oxidative stress produced by amyloid beta. PMID- 12770557 TI - Mother's voice "buffers" separation-induced receptor changes in the prefrontal cortex of octodon degus. AB - Although the potential vulnerability of the postnatally developing brain toward adverse environmental influences is generally recognized, relatively little is known about the basic mechanisms involved. The plasticity and adaptability of the postnatally developing brain in response to adverse emotional experiences was analyzed in the South American Octodon degus. Our study revealed that repeated brief separation from the parents and exposure to an unfamiliar environment induces an up-regulation of dopamine (D1) and 5-hydroxytrytamine (5HT1(A)) receptor density in the precentral medial, anterior cingulate, prelimbic and infralimbic cortices in female pups. No significant changes of gamma aminobutyric acid (GABA(A)) receptor density were found in deprived animals of both genders. The acoustic presence of the mother during parental separation suppressed the D1 receptor up-regulation as well as the 5-HT1(A)-receptor up-regulation, again only in the female pups. These results demonstrate that that early adverse emotional experience alters aminergic function within the prefrontal cortex in the female but not the male brain. The mother's voice, a powerful emotional signal, can protect the developing cortex from separation-induced receptor changes. PMID- 12770558 TI - Responses of rat trigeminal neurones to dental pulp cells or fibroblasts overexpressing neurotrophic factors in vitro. AB - The adult dental pulp is innervated by sensory trigeminal axons and efferent sympathetic axons. Rat trigeminal ganglia extend neurites when co-cultivated in vitro with pulpal tissue explants, suggesting that pulpal cells secrete soluble molecules that stimulate the growth of trigeminal ganglion axons. In addition, cultured pulpal cells produce mRNAs for neurotrophins and glial cell line-derived neurotrophic factor-family members. These data suggest that neurotrophic factors are involved in the formation of a pulpal innervation. Here, we examine how pulpal cells and 3T3 fibroblasts overexpressing certain neurotrophic factors (nerve growth factor, brain-derived neurotrophic factor, neurotrophin-3, neurotrophin-4, glial cell line-derived neurotrophic factor or neurturin) influence survival and growth of single trigeminal ganglion neurones in vitro in quantitative terms. The results show that most of the neurotrophic factor overexpressing fibroblasts induce similar neuronal soma diameters, but higher survival rates and neurite lengths compared with pulpal cells. With respect to neurite growth pattern, trigeminal ganglion neurones co-cultured with fibroblasts overexpressing nerve growth factor develop a geometry that is most similar to that seen in co-cultures with pulpal cells. We conclude that none of the fibroblasts overexpressing neurotrophic factors can fully mimic the effects of pulpal cells on trigeminal ganglion neurones, and that nerve growth factor promotes a neurite growth pattern most similar to the picture seen in co-cultures with pulpal cells. PMID- 12770560 TI - Distribution and colocalisation of glutamate decarboxylase isoforms in the rat spinal cord. AB - The inhibitory neurotransmitter GABA is synthesized by glutamic acid decarboxylase (GAD), and two isoforms of this enzyme exist: GAD65 and GAD67. Immunocytochemical studies of the spinal cord have shown that whilst both are present in the dorsal horn, GAD67 is the predominant form in the ventral horn. The present study was carried out to determine the pattern of coexistence of the two GAD isoforms in axonal boutons in different laminae of the cord, and also to examine the relation of the GADs to the glycine transporter GLYT2 (a marker for glycinergic axons), since many spinal neurons are thought to use GABA and glycine as co-transmitters. Virtually all GAD-immunoreactive boutons throughout the spinal grey matter were labelled by both GAD65 and GAD67 antibodies; however, the relative intensity of staining with the two antibodies varied considerably. In the ventral horn, most immunoreactive boutons showed much stronger labelling with the GAD67 antibody, and many of these were also GLYT2 immunoreactive. However, clusters of boutons with high levels of GAD65 immunoreactivity were observed in the motor nuclei, and these were not labelled with the GLYT2 antibody. In the dorsal horn, some GAD-immunoreactive boutons had relatively high levels of labelling with either GAD65 or GAD67 antibody, whilst others showed a similar degree of labelling with both antibodies. GLYT2 immunoreactivity was associated with many GAD-immunoreactive boutons; however, this did not appear to be related to the pattern of GAD expression. It has recently been reported that there is selective depletion of GAD65, accompanied by a loss of GABAergic inhibition, in the ipsilateral dorsal horn in rats that have undergone peripheral nerve injuries [J Neurosci 22 (2002) 6724]. Our finding that some boutons in the superficial laminae showed relatively high levels of GAD65 and low levels of GAD67 immunoreactivity is therefore significant, since a reduction in GABA synthesis in these axons may contribute to neuropathic pain. PMID- 12770559 TI - Dual modulation of gabaergic transmission by metabotropic glutamate receptors in rat ventral tegmental area. AB - The effects of metabotropic glutamate receptor (mGluR) activation on non-dopamine (putative GABAergic) neurons and inhibitory synaptic transmission in the ventral tegmental area were examined using intracellular recordings from rat midbrain slices. Perfusion of (+/-)-1-aminocyclopentane-trans-1,3-dicarboxylic acid (t ACPD; agonist for group I and II mGluRs), but not L-amino-4-phosphonobutyric acid (L-AP4; agonist for group III mGluRs), produced membrane depolarization (current clamp) and inward current (voltage clamp) in non-dopamine neurons. The t-ACPD induced depolarization was concentration-dependent (concentration producing 50% maximal depolarization [EC(50)]=6.1+/-2.5 microM), and was blocked by the antagonist (+/-)-alpha-methyl-4-carboxyphenylglycine, but not by tetrodotoxin and ionotropic glutamate-receptor antagonists. The t-ACPD-evoked responses were mimicked comparably by selective group I mGluR agonist (RS)-3,5 dihydroxyphenylglycine (DHPG). Furthermore, the DHPG-induced depolarization in non-dopamine neurons was greatly reduced by mGluR1-specific antagonist 7(hydroxyimino)cyclopropachromen-1a-carboxylate ethyl ester. When recorded in dopamine neurons, the frequency of spontaneous GABA(A) receptor-mediated inhibitory postsynaptic potentials was increased by t-ACPD but not L-AP4. However, the amplitude of evoked inhibitory postsynaptic currents in dopamine neurons was reduced by all three group mGluR agonists. These results reveal a dual modulation of mGLuR activation on inhibitory transmission in midbrain ventral tegmental area: enhancing putative GABAergic neuronal excitability and thus potentiating tonic inhibitory synaptic transmission while reducing evoked synaptic transmission at inhibitory terminals. PMID- 12770561 TI - Involvement of beta-adrenergic receptors in protein synthesis-dependent late long term potentiation (LTP) in the dentate gyrus of freely moving rats: the critical role of the LTP induction strength. AB - We have investigated the requirement of beta-adrenergic receptor activation and protein synthesis for the induction and specifically for the maintenance of long term potentiation (LTP) in the dentate gyrus of freely moving rats in dependency on different LTP-induction procedures. Three tetanization paradigms were used: a relatively weak protocol A (10 bursts of 15 biphasic pulses at 200 Hz; 10-s interburst interval; 0.2-ms pulse width per phase), a stronger protocol B (as protocol A but 20 bursts and 0.25-ms pulse width) and, as the strongest condition, protocol C (2 times protocol B; inter-tetanus interval: 5 min). All protocols led to robust late-LTP in control animals. Late- but not early-LTP was protein synthesis-dependent under all tetanization conditions as indicated by the absence of long-lasting LTP when the protein synthesis inhibitor anisomycin was applied before tetanization. Application of the beta-adrenergic receptor antagonist propranolol before LTP induction prevented late-LTP when either protocol A or B but not when protocol C was used. Thus, repeated strong tetanization can compensate for the loss of beta-adrenergic receptor activation. We suggest that the results could provide a link to cellular mechanisms of memory consolidation in respect to the strength and relevance of the incoming sensory information during learning. PMID- 12770562 TI - Comparative analysis of fatty acid amide hydrolase and cb(1) cannabinoid receptor expression in the mouse brain: evidence of a widespread role for fatty acid amide hydrolase in regulation of endocannabinoid signaling. AB - Fatty acid amide hydrolase (FAAH) catalyses hydrolysis of the endocannabinoid arachidonoylethanolamide ("anandamide") in vitro and regulates anandamide levels in the brain. In the cerebellar cortex, hippocampus and neocortex of the rat brain, FAAH is located in the somata and dendrites of neurons that are postsynaptic to axon fibers expressing the CB(1) cannabinoid receptor [Proc R Soc Lond B 265 (1998) 2081]. This complementary pattern of FAAH and CB(1) expression provided the basis for a hypothesis that endocannabinoids may function as retrograde signaling molecules at synapses in the brain [Proc R Soc Lond B 265 (1998) 2081; Phil Trans R Soc Lond 356 (2001) 381] and subsequent experimental studies have confirmed this [Science 296 (2002) 678]. To assess more widely the functions of FAAH in the brain and the potential impact of FAAH activity on the spatiotemporal dynamics of endocannabinoid signaling in different regions of the brain, here we have employed immunocytochemistry to compare the distribution of FAAH and CB(1) throughout the mouse brain, using FAAH(-/-) mice as negative controls to validate the specificity of FAAH-immunoreactivity observed in wild type animals. In many regions of the brain, a complementary pattern of FAAH and CB(1) expression was observed, with FAAH-immunoreactive neuronal somata and dendrites surrounded by CB(1)-immunoreactive fibers. In these regions of the brain, FAAH may regulate postsynaptic formation of anandamide, thereby influencing the spatiotemporal dynamics of retrograde endocannabinoid signaling. However, in some regions of the brain such as the globus pallidus and substantia nigra pars reticulata, CB(1) receptors are abundant but with little or no associated FAAH expression and in these brain regions the spatial impact and/or duration of endocannabinoid signaling may be less restricted than in regions enriched with FAAH. A more complex situation arises in several regions of the brain where both FAAH and CB(1) are expressed but in a non-complementary pattern, with FAAH located in neurons and/or oligodendrocytes that are proximal but not postsynaptic to CB(1)-expressing axon fibers. Here FAAH may nevertheless influence endocannabinoid signaling but more remotely. Finally, there are regions of the brain where FAAH-immunoreactive neurons and/or oligodendrocytes occur in the absence of CB(1)-immunoreactive fibers and here FAAH may be involved in regulation of signaling mediated by other endocannabinoid receptors or by receptors for other fatty acid amide signaling molecules. In conclusion, by comparing the distribution of FAAH and CB(1) in the mouse brain, we have provided a neuroanatomical framework for comparative analysis of the role of FAAH in regulation of the spatiotemporal dynamics of retrograde endocannabinoid signaling in different regions of the brain. PMID- 12770563 TI - Manipulation of dopamine d1-like receptor activation in the rat medial prefrontal cortex alters stress- and cocaine-induced reinstatement of conditioned place preference behavior. AB - These studies examined the ability of the dopamine D1-like agonist SKF 81297 and D1-like antagonist SCH 23390 in the medial prefrontal cortex to alter the reinstatement of cocaine-induced conditioned place preference behavior. Male Sprague-Dawley rats were fitted with bilateral cannulae over the medial prefrontal cortex and subsequently trained in a conditioned place preference task. Animals were trained in this task using four pairings of cocaine (12 mg/kg, i.p.). Conditioned place preference was demonstrated in all animals, and this behavior was then extinguished over a 5-10-day period before testing for reinstatement. Just prior to reinstatement by immobilization stress or a cocaine priming injection (5 mg/kg, i.p.), a microinjection of the D1-like receptor antagonist SCH 23390 (0.01, 0.1 or 1.0 microg/side), or the D1-like receptor agonist SKF 81297 (0.1, 0.3 or 1.0 microg/side) was given into the medial prefrontal cortex. SCH 23390 blocked both stress- and cocaine-induced reinstatement of conditioned place preference after the two higher doses were administered into the medial prefrontal cortex. The highest dose of SKF 81297 (1.0 microg/side) prevented immobilization stress- but not cocaine-induced reinstatement. The highest dose of these drugs given in the absence of stress or cocaine did not produce reinstatement. The results indicate that immobilization stress given within the place-preference chamber is capable of producing reinstatement of cocaine-seeking behavior. The microinjection studies suggest that D1-like receptor antagonism within the prefrontal cortex is sufficient to block reinstatement by stress and cocaine. Furthermore, the results from D1-like receptor activation in the medial prefrontal cortex point to utilization of different neural pathways for stress- and cocaine-induced reinstatement. PMID- 12770565 TI - Differential activation in the medial temporal lobe during a sound-sequence discrimination task across age in human subjects. AB - To elucidate the brain mechanisms to encode sequential events, event-related potentials (ERPs) were recorded during a sound-sequence discrimination task using young and middle-aged adult subjects. In the task, a series of six or 12 kinds of natural sounds were sequentially presented; 70-80% of the stimuli were presented in a fixed order (Non-target), but the remaining stimuli, in a random order (Target). The subjects were instructed to detect the Targets and press a button at the end of each Target. In a control task, the same sounds were randomly presented (Control), and they were instructed to press the button at the end of each sound. Behavioral results indicated that the young subjects learned the task faster than did the middle-aged subjects. Positive ERP waves were evoked by Targets and Non-targets in the parieto-occipital area around 300-700 ms after stimulus onset. The mean amplitudes during this period in the young subjects were larger in Target than Control conditions, and those in Target condition were larger in the young than middle-aged subjects. Furthermore, the mean amplitudes in the Target condition were significantly correlated with behavioral performance. Equivalent dipoles for the ERPs evoked by Targets were estimated in the medial temporal lobe including the hippocampal formation and parahippocampal gyrus. The results suggest that the ERPs around 300-700 ms latency are involved in sound-sequence information processing. Furthermore, decrease in amplitudes of this positivity in the middle-aged subjects suggests that age-related memory decline is associated with deficits in encoding and retrieval of unfamiliar sequence. PMID- 12770564 TI - Deprivation of sensory inputs to the olfactory bulb up-regulates cell death and proliferation in the subventricular zone of adult mice. AB - The main olfactory bulb (MOB) is the first relay on the olfactory sensory pathway and the target of the neural progenitor cells generated in the subventricular zone (SVZ) lining the lateral ventricles and which migrate along the rostral extension of the SVZ, also called the rostral migratory stream (RMS). Within the MOB, the neuroblasts differentiate into granular and periglomerular interneurons. A reduction in the number of granule cells during sensory deprivation suggests that neurogenesis may be influenced by afferent activity. Here, we show that unilateral sensory deafferentation of the MOB by axotomy of the olfactory receptor neurons increases apoptotic cell death in the SVZ and along the rostro caudal extent of the RMS. The vast majority of dying cells in the RMS are migrating neuroblasts as indicated by double Terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick-end labeling/PSA-NCAM labeling. Counting bromodeoxyuridine-labeled cells in animals killed immediately or 4 days after tracer administration showed a bilateral increase in proliferation in the SVZ and RMS which was balanced by cell death on the operated side. These data suggest that olfactory inputs are required for the survival of newborn neural progenitors. The greatest enhancement in proliferation occurred in the extension of the RMS located in the MOB, revealing a population of local precursors mitotically stimulated following axotomy. Together, these findings indicate that olfactory inputs may strongly modulate the balance between neurogenesis and apoptosis in the SVZ and RMS and provide a model for further investigation of the underlying molecular mechanisms of this activity-dependent neuronal plasticity. PMID- 12770566 TI - Hippocampal cell proliferation and epileptogenesis after audiogenic kindling are not accompanied by mossy fiber sprouting or Fluoro-Jade staining. AB - Repetitive sound-induced seizures, known as audiogenic kindling (AK), gradually induce the transference of epileptic activity from brainstem to forebrain structures along with behavioral changes. The aim of our work was to correlate the behavioral changes observed during the AK with possible alterations in neuronal proliferation, cell death, hippocampal mossy fiber sprouting and in the EEG pattern of Wistar audiogenic rats, a genetically susceptible strain from our laboratory. Susceptible and non-susceptible animals were submitted to repeated sound stimulations for 14-16 days and hippocampal mitotic activity was studied through the incorporation of bromodeoxyuridine (BrdU). Cell death and mossy fiber sprouting were assessed, respectively, by using Fluoro-Jade and Timm staining, 2 and 32 days after the last kindling stimulation. In addition, we used immunofluorescent double labeling for a glial and a mitotic marker to evaluate newly born cell identity. Some animals had hippocampus and amygdala electrodes for EEG recordings. Our results show that kindled animals with 6-11 generalized limbic seizures (class IV-V) had increased cell proliferation in the dentate gyrus when compared with animals with zero or one to three seizures. BrdU positive cells labeled on day 2 and on day 32 were both GFAP negative. In the later group, rounded and well-defined BrdU-positive/GFAP-negative nuclei were seen in different portions of the granule cell layer. We did not observe any Fluoro-Jade or differential Timm staining in kindled animals at both killing times. However, EEG recordings showed intense epileptic activity in the hippocampus and amygdala of all animals with limbic seizures.Therefore, our data indicate that AK-induced limbic epileptogenicity is able to increase the hippocampal mitotic rate, even though it does not seem to promote neuronal death or mossy fiber sprouting in the supragranular layer of the dentate gyrus. PMID- 12770567 TI - Central administration of the neurotensin receptor antagonist SR48692 attenuates vacuous chewing movements in a rodent model of tardive dyskinesia. AB - Tardive dyskinesia is a movement disorder that develops in 20-30% of patients treated with chronic neuroleptics. Whilst the pathogenesis of tardive dyskinesia remains unclear, altered expression of neuropeptides in the basal ganglia has been implicated in its emergence. The peptide neurotensin is expressed in both dopamine D1 receptor-bearing neurons of the direct striatonigral pathway and dopamine D2 receptor-bearing neurons of the indirect striatopallidal pathway. Increased levels of striatal neurotensin messenger RNA (mRNA) are reported following chronic neuroleptic therapy. Chronic treatment with the typical antipsychotic haloperidol elicits neurotensin immunoreactivity in a large number of striatopallidal and a modest number of striatonigral projection neurons, whilst treatment with the potent dopamine releaser, methamphetamine, induces intense neurotensin immunoreactivity in striatonigral projection neurons. In order to determine whether increased levels of striatal neurotensin mRNA in the direct striatonigral or the indirect striatopallidal pathway play a more influential role in the development of tardive dyskinesia, we explored the effects of a specific neurotensin antagonist in a rodent model (vacuous chewing movements [VCMs] induced by chronic neuroleptics). Three groups of animals received injections of fluphenazine decanoate (25 mg/kg) or its vehicle sesame oil every 3 weeks for at least 18 weeks. They were then surgically implanted with bilateral guide cannulae aimed at the striatum, the substantia nigra pars reticulata, or the globus pallidus respectively. After recovery, animals were infused with 2-[(1-(7-chloro-4-quinolinyl)-5-(2,6-imethoxyphenyl)pyrazol-3 yl)carbonylamino]tricyclo(3.3.1.1.(3.7))decan-2-carboxylic acid (SR48692; 0.25, 0.50, and 1.0 nmol/microl), or its vehicle (10% dimethyl sulfoxide [DMSO] in saline) and observed for 60 min. Intra-striatal, intra-nigral or intra-pallidal infusion of SR48692 attenuated neuroleptic-induced VCMs. These findings lend further support to a role for neurotensin in the development of VCMs but do not clarify which pathway plays a more important role. Thus, treatments that reduce or prevent the effects of increased neurotensin expression and release may be useful in the management of tardive dyskinesia. PMID- 12770568 TI - Glutamate release in the nucleus accumbens is involved in behavioral depression during the PORSOLT swim test. AB - An abnormality in glutamate function has been implicated in the neural substrate of depressive disorders. To investigate this in rats, the Porsolt swim test was used to assess the role of glutamate in the nucleus accumbens. Glutamate injected into the nucleus accumbens dose-dependently decreased swimming time on the test day (day 2), whereas N-methyl-D-aspartate antagonists dizocilpine and 2-amino-5 phosphonovalerate increased swimming, like an antidepressant. Dizocilpine injected before the conditioning trial (day 1) did not modify the swimming times during the first day but abolished behavioral depression on day 2. Microdialysis coupled to capillary-zone electrophoresis was then used to determine in vivo changes in glutamate release in 1-min samples during the swim test. On day 1, glutamate increased significantly and reached a maximum of 222% after 3 min of swimming. On day 2, baseline glutamate levels were back to normal, but when the animal was placed in the water, glutamate increased to 419% during the first minute, and the animals swam significantly less. For comparison, tail pinch on consecutive days was used as a nonspecific, repeated stressor while accumbens glutamate levels were measured. Tail pinch on the first day increased glutamate similar to the effect obtained during the first day of swimming; however, a second day of tail pinch decreased glutamate levels, instead of the potentiated response observed during the second day of swimming. These results show that accumbens glutamate plays a role in causing the behavioral aspects of depressed behavior as modeled in the swim test. The accumbens may be a potential site of action for drugs that alter behavioral depression. PMID- 12770569 TI - Serotonergic serotonin (1A) mixed agonists/antagonists elicit large-magnitude phase shifts in hamster circadian wheel-running rhythms. AB - The biological clock that generates circadian rhythms in mammals is located within the suprachiasmatic nuclei at the base of the hypothalamus. The circadian clock is entrained to the daily light/dark cycle by photic information from the retina. The retinal input to the clock is inhibited by exogenously applied serotonin agonists, perhaps mimicking an endogenous inhibitory serotonergic input to the clock arriving from the midbrain raphe. In the present study, a unique class of serotonergic compounds was tested for its ability to modulate retinal input to the circadian clock. The serotonergic ligands 8-(2-[4-(2-methoxyphenyl) 1-piperazinyl]ethyl)-8-azaspiro(4.5)decane-7,9-dione dihydrochloride (BMY 7378), S 15535, and 8-[2-(1,4-benzodioxan-2-ylmethylamino)ethyl]-8-azaspiro[4.5]decane 7,9-dione hydrochloride (MDL 73005 EF) can all be classified as mixed agonists/antagonists at type 1A serotonin receptors. Circadian wheel-running activity rhythms were monitored in Syrian hamsters maintained in constant darkness. Dim white-light pulses administered to the hamsters at circadian time 19 advanced the phase of their running rhythms by 1-2 h. Injection of BMY 7378, S 15535, and to a lesser degree MDL 73005 EF, prior to the light pulses resulted in phase advances from 5 to 6 h, and by as much as 8 h. Neither BMY 7378 nor S 15535 had any effect on light-induced phase delays in hamster activity rhythms at circadian time 14. Further, BMY 7378 is able to phase advance circadian rhythms by approximately 1 h at night even without light exposure. Finally, the effects of BMY 7378 on circadian rhythms is opposite to that observed with the prototypical serotonin 1A agonist (+/-)-8-hydroxy-2-(DI-n-propyl-amino)tetralin hydrobromide (8-OH-DPAT) (8-OH-DPAT elicits non-photic phase advances in the day and inhibits photic-induced phase advances at night). These results suggest that pharmacologically blocking raphe input to the suprachiasmatic circadian clock results in substantially larger photically induced phase advances in wheel running rhythms. This is further evidence that raphe input to the circadian clock is probably acting to dampen the clock's response to light under certain conditions. The large-magnitude phase shifts, and temporal-activity profile seen with BMY 7378 and S 15535, suggest that compounds with this unique pharmacological profile may be beneficial in the treatment of circadian phase delays recently reported to be a complication resulting from Alzheimer's disease. PMID- 12770570 TI - Positional firing properties of postrhinal cortex neurons. AB - Hippocampal cell firing in awake, behaving rats is often spatially selective, and such cells have been called place cells. Similar spatial correlates have also been described for neurons in the medial entorhinal and perirhinal cortices. All three regions receive sensory associational input from postrhinal cortex, which, in turn, is heavily interconnected with visuospatial neocortical regions. The spatial selectivity of postrhinal cells, however, has never been examined. Here, we report the activity of neurons in postrhinal cortex of freely moving rats performing a spatial task on a four-arm radial maze. Data are also reported for visual association cortex neurons. The four-arm radial maze was defined by multisensory cues on the surfaces of the maze arms (proximal) and complex visual cues at the surround (distal). On each recording day, rats were run in three conditions: baseline, double cue rotation (proximal +90 degrees; distal -90 degrees ), and baseline. In this task, hippocampal place field activity is robust and can be controlled by proximal or distal cues. The majority of postrhinal neurons (64%) exhibited positional correlates during performance on the task; however, characteristics of these postrhinal cells were substantially different from those previously described for hippocampal place cells. Most postrhinal cells with firing fields exhibited split or multiple subfields (93%). Unlike hippocampal place fields, the large majority of postrhinal firing fields (84%) adopted new spatial correlates when experimental cues were rotated, but did so neither predictably nor concordantly. This is the first report of positional firing correlates in the postrhinal cortex. The data are consistent with the idea that postrhinal cortex participates in visuospatial functions by monitoring changes in environmental stimuli rather than encoding stable spatial cues. Thus, postrhinal neurons appear to participate in higher-level perceptual functions rather than mnemonic functions. We propose that the response properties of postrhinal neurons represent an early step in a spatial pathway that culminates in the specific and stable place fields of the hippocampus. PMID- 12770572 TI - Electrophysiological evidence for the existence of a posterior cortical prefrontal-basal forebrain circuitry in modulating sensory responses in visual and somatosensory rat cortical areas. AB - The prefrontal cortex (PFC) receives input from sensory neocortical regions and sends projections to the basal forebrain (BF). The present study tested the possibility that pathways from sensory cortical regions via the PFC-BF and from the BF back to specific sensory cortical areas could modulate sensory responses. Two prefrontal areas that responded to stimulation of the primary somatosensory and visual cortices were delineated: an area encompassing the rostral part of the cingulate cortex that responded to visual cortex stimulation, and a region dorso lateral to the first in the precentral-motor association area that reacted to somatosensory cortex stimulation. Moreover, BF neurons responded to PFC electrical stimulation. They were located in the ventral pallidum, substantia innominata and the horizontal limb of the diagonal-band areas. Of the responsive BF neurons 42% reacted only to stimulation of 'visually-responsive,' 33% responded only to the 'somatosensory-responsive' prefrontal sites and the remaining neurons reacted to both prefrontal cortical areas. The effect of BF and PFC stimulations on somatosensory and visual-evoked potentials was tested. BF stimulation increased the amplitude of both sensory-evoked potentials. However, stimulation of the 'somatosensory-responsive' prefrontal area increased only somatosensory-evoked potentials while 'visually-responsive' prefrontal-area stimulation increased only visual-evoked potentials. Atropine blocked both facilitatory effects. The proposed cortico-prefronto-basalo-cortical circuitry may have an important role in cortical plasticity and selective attention. PMID- 12770571 TI - Regulation of the developing hypothalamic-pituitary-adrenal axis in corticotropin releasing hormone receptor 1-deficient mice. AB - During postnatal development, mice undergo a so-called stress hyporesponsive period, which is characterized by low basal corticosterone levels and the inability of mild stressors to induce a corticosterone response. The stress hyporesponsiveness is in part regulated by maternal factors. Twenty-four hours of deprivation results in an activation of basal and stress-induced corticosterone and a down-regulation of corticotropin releasing hormone (CRH), mineralocorticoid receptor (MR) and glucocorticoid receptor (GR) expression in the brain. It has been hypothesized that the CRH receptor 1 (CRHr1) may play an important regulatory role during development by mediating the effects of maternal deprivation. Using CRHr1-deficient mice we examined the role of this receptor on the maternal deprivation effects and in regulating the expression of hypothalamic pituitary-adrenal axis-related genes. We could demonstrate that the CRHr1 is essential for the activation of the corticosterone response following maternal deprivation, most likely due to the lack of the receptor in the pituitary. Furthermore, we could show that the CRHr1 is regulating the expression of CRH and MRs. In contrast, effects of maternal deprivation during postnatal development on GRs are not mediated by this receptor. PMID- 12770573 TI - Attenuation of circadian light induced phase advances and delays by neuropeptide Y and a neuropeptide Y Y1/Y5 receptor agonist. AB - Circadian rhythms can be synchronised to photic and non-photic stimuli. The circadian clock, anatomically defined as the suprachiasmatic nucleus in mammals, can be phase shifted by light during the night. Non-photic stimuli reset the circadian rhythm during the day. Photic and non-photic stimuli have been shown to interact during the day and night. Precise mechanisms for these complex interactions are unknown. A possible pathway for non-photic resetting of the clock is thought to generate from the intergeniculate leaflet, which conveys information to the suprachiasmatic nucleus (SCN) through the geniculohypothalamic tract and utilises neuropeptide Y (NPY) as its primary neurotransmitter. Interactions between light and NPY were investigated during the early (2 h after activity onset) and late (6 h after activity onset) night in male Syrian hamsters. NPY microinjections into the region of the SCN significantly attenuated light-induced phase delay, during the early subjective night. Phase advances to light were completely inhibited by the administration of NPY during the late night. The precise mechanism by which NPY attenuates or blocks photic phase shifts is unclear, but the NPY Y5 receptor has been implicated in the mediation of this inhibitory effect. The NPY Y1/Y5 receptor agonist, [Leu(31),Pro(34)]NPY, was administered via cannula microinjections following light exposure during the early and late night. [Leu(31),Pro(34)]NPY significantly attenuated phase delays to light during the early night and blocked phase advances during the late night, in a manner similar to NPY. These results show the ability of NPY to attenuate phase shifts to light during the early night and block light-induced phase advances during the late night. Furthermore, this is the first in vivo study implicating the involvement of the NPY Y1/Y5 receptors in the complex interaction of photic and non-photic stimuli during the night. The alteration of photic phase shifts by NPY may influence photic entrainment within the circadian system. PMID- 12770575 TI - The molecular biology of knockdown resistance to pyrethroid insecticides. AB - The term "knockdown resistance" is used to describe cases of resistance to diphenylethane (e.g. DDT) and pyrethroid insecticides in insects and other arthropods that result from reduced sensitivity of the nervous system. Knockdown resistance, first identified and characterized in the house fly (Musca domestica) in the 1950's, remains a threat to the continued usefulness of pyrethroids in the control of many pest species. Research since 1990 has provided a wealth of new information on the molecular basis of knockdown resistance. This paper reviews these recent developments with emphasis on the results of genetic linkage analyses, the identification of gene mutations associated with knockdown resistance, and the functional characterization of resistance-associated mutations. Results of these studies identify voltage-sensitive sodium channel genes orthologous to the para gene of Drosophila melanogaster as the site of multiple knockdown resistance mutations and define the molecular mechanisms by which these mutations cause pyrethroid resistance. These results also provide new insight into the mechanisms by which pyrethroids modify the function of voltage sensitive sodium channels. PMID- 12770576 TI - cDNA cloning, purification, properties, and function of a beta-1,3-glucan recognition protein from a pyralid moth, Plodia interpunctella. AB - Microorganisms possess distinctive biochemical or molecular patterns on their cell surfaces, such as those formed by the lipopolysaccharides, lipoteichoic acids, and/or peptidoglycans of bacteria and the beta-1,3-glucans of fungi. Pattern recognition proteins that bind to these surface moieties have been implicated in the activation of the innate immune response in insects and other invertebrates. We report the purification and cloning of a cDNA for a 53-kDa beta 1,3-glucan recognition protein (betaGRP) from the Indianmeal moth, Plodia interpunctella (Hubner) (Lepidoptera: Pyralidae). BetaGRP cDNA contains an open reading frame that encodes 488 amino acids, of which the first 17 residues comprise the secretion signal peptide. The calculated molecular mass of the 471 residue mature protein is 53,311 Da. The protein consists of a carboxyl-terminal domain that is similar to other recognition proteins from invertebrates, beta-1,3 glucanases from bacteria, and a beta-1,3-glucanase from the sea urchin, Strongylocentrotus purpuratus. The amino-terminus of betaGRP shares sequence similarity with other invertebrate recognition molecules and the beta-1,3 glucanase from S. purpuratus. Affinity purification of a 53-kDa protein and subsequent sequencing of a peptide produced by tryptic cleavage confirmed the presence of the betaGRP in P. interpunctella larval hemolymph. RT-PCR analysis indicates that betaGRP is constitutively expressed in all life-stages, with no detectable induction following exposure of wandering larvae to microbial elicitors. Northern blot analysis indicates that the 1.8-kb betaGRP transcript is transcribed within the fat body. Recombinant betaGRP retains beta-1,3-glucan binding activity, binds to lipopolysaccharide and lipoteichoic acid in vitro, causes aggregation of microorganisms, and activates the prophenoloxidase cascade in the presence of soluble beta-1,3-glucan. These data support the hypothesis that the 53-kDa betaGRP functions to recognize pathogen surface molecules as nonself and subsequently activates insect innate immune responses. PMID- 12770578 TI - Juvenile hormone biosynthesis in moths: synthesis and evaluation of farnesol homologs as alternate substrates of farnesol oxidase. AB - The oxidation of farnesol to farnesal is an important step in insect juvenile hormone (JH) biosynthesis and is mediated by one or more alcohol oxidases located within the minute endocrine gland, the corpus allatum. Because lepidopteran insects have the capacity to produce homologous JH structures, the substrate selectivity of farnesol oxidase was examined by determining the ability of several terpenol homologs to inhibit farnesol oxidation in moths. Results utilizing corpora allata homogenates from larval, adult, and embryonic Manduca sexta indicate that increased steric bulk at the C-3 position of the sesquiterpenol chain is detrimental to inhibitory potency. Triethylhomofarnesol (1h), which is precursor to JH 0 and therefore a physiologically important metabolite of M. sexta embryos, was found to be a poor inhibitor of farnesol oxidation but was oxidized in almost same amount as farnesol. This data indicate that farnesol oxidase of the corpus allatum plays a limited role in controlling JH homolog production in moths, and suggests that another oxidative enzyme, which is present at early stages of moth development, is involved in JH homolog construction. PMID- 12770577 TI - RdlDv, a novel GABA-gated chloride channel gene from the American dog tick Dermacentor variabilis. AB - The American dog tick Dermacentor variabilis is a major transmitter of bacterial and viral pathogens in human and animal populations, and compounds active against this species would benefit both human and animal health. Invertebrate GABA-gated chloride channels are validated targets of commonly used insecticides and acaricides. We cloned a novel member of the invertebrate GABA-gated chloride channel gene family from Dermacentor variabilis, RdlDv. The closest homologue of the predicted gene product of RdlDv is the RDL protein encoded by the GABA-gated chloride channel gene Drosophila Rdl (Resistance to Dieldrin), with which it shares 64% amino acid identity. When expressed in Xenopus oocytes, RdlDv produces GABA-activated currents blocked by the known insecticides and RDL antagonists fipronil and picrotoxinin. These results suggest that RdlDv encodes a GABA-gated chloride channel subunit, making it a potential target for compounds active against the tick D. variabilis. PMID- 12770579 TI - Multiple acyl-CoA desaturase-encoding transcripts in pheromone glands of Helicoverpa assulta, the oriental tobacco budworm. AB - Seven desaturase cDNAs were isolated from pheromone glands of Helicoverpa assulta, a moth producing a sex pheromone blend with high Z9-16:Ald and low Z11 16:Ald, opposite to what is found in other heliothine moths such as Helicoverpa zea. Six of the seven sequences map onto recently defined lepidopteran desaturase sequence lineages and the other is orthologous to a desaturase sequence previously reported only in H. zea. The levels of desaturase-encoding transcripts in pheromone glands were determined and the three most abundant ones were functionally expressed in a desaturase-deficient mutant strain of Saccharomyces cerevisiae. The HassNPVE transcript, shown to encode a delta9 desaturase producing more Z9-18:Acid than Z9-16:Acid, was the most abundant, followed by the HassKPSE transcript, shown to encode a delta9 desaturase producing more Z9 16:Acid than Z9-18:Acid, and by the HassLPAQ transcript, shown to encode a delta11 desaturase producing only Z11-16:Acid. Thus, the relative amounts of transcripts encoding two delta9 desaturases and a single delta11 desaturase in H. assulta pheromone glands were consistent with the relative amounts of unsaturated fatty acid precursors required to produce the major and minor sex pheromone components of this species. Desaturase transcript levels in pheromone glands were also found to be as high during scotophase as during light phase, when pheromone production ceases. The other four transcripts were present at extremely low levels in H. assulta pheromone glands and the functional roles of their encoded desaturase-homologous proteins could not be determined. PMID- 12770580 TI - The influence of tick behavior, biotope and host specificity on concerted evolution of the platelet aggregation inhibitor savignygrin, from the soft tick Ornithodoros savignyi. AB - Ticks are obligate blood-feeding parasites that secrete anti-hemostatic components during feeding to enable control of the hemostatic system of the host. Complex interactions at the tick-host interface are an indication of the important role that the host played during tick evolution. The question is to what extent interaction with the host and the environment influences tick evolution. Previously, two isoforms (97% sequence identity) of savignygrin, an alphaIIbbeta3 antagonist, have been described. The presence of both isoforms within 20 random individuals confirmed that these isoforms must be recent gene duplicates. Analysis of the sequence differences between the isoforms shows a Kn/Ks ratio of 1, which indicates neutral selection for the isoforms. However, the biased localization of differences within the 3' end of the genes suggests that concerted evolution acts on the isoforms. Calculation of the divergence date between the isoforms (1.6-5.2 MYA) also indicates purifying selection, as ample time had passed after duplication, for inactivation of one gene copy. We conclude that concerted evolution has functioned to maintain a high copy number of the savignygrins in order for Ornithodoros savignyi to parasitize a wide host range. This contrasts with O. moubata that expresses the savignygrin homolog, disagregin, as a single copy at lower concentration levels and correlates with the confined habitat and consequently narrow host range of O. moubata. Recent "domestication" of O. savignyi due to animal husbandry practices could however, have reduced the selection constraints acting to maintain the gene copies as evidenced by the structural instability of one of the isoforms. Our results suggest that environmental factors and host associations do play an important role in the evolution of anti-hemostatic components in ticks. PMID- 12770581 TI - Properties of catalytic, linker and chitin-binding domains of insect chitinase. AB - Manduca sexta (tobacco hornworm) chitinase is a glycoprotein that consists of an N-terminal catalytic domain, a Ser/Thr-rich linker region, and a C-terminal chitin-binding domain. To delineate the properties of these domains, we have generated truncated forms of chitinase, which were expressed in insect cells using baculovirus vectors. Three additional recombinant proteins composed of the catalytic domain fused with one or two insect or plant chitin-binding domains (CBDs) were also generated and characterized. The catalytic and chitin-binding activities are independent of each other because each activity is functional separately. When attached to the catalytic domain, the CBD enhanced activity toward the insoluble polymer but not the soluble chitin oligosaccharide primarily through an effect on the Km for the former substrate. The linker region, which connects the two domains, facilitates secretion from the cell and helps to stabilize the enzyme in the presence of gut proteolytic enzymes. The linker region is extensively modified by O-glycosylation and the catalytic domain is moderately N-glycosylated. Immunological studies indicated that the linker region, along with elements of the CBD, is a major immunogenic epitope. The results support the hypothesis that the domain structure of insect chitinase evolved for efficient degradation of the insoluble polysaccharide to soluble oligosaccharides during the molting process. PMID- 12770582 TI - Sterol biosynthesis by symbiotes: cytochrome P450 sterol C-22 desaturase genes from yeastlike symbiotes of rice planthoppers and anobiid beetles. AB - Rice planthoppers and anobiid beetles harbor intracellular yeastlike symbiotes (YLS), whose sterols are nutritionally advantageous for the host insects that cannot synthesize sterols. YLS of anobiid beetles synthesize ergosterol, whereas YLS of planthoppers produce ergosta-5,7,24(28)-trienol, which is a metabolic intermediate in the ergosterol biosynthetic pathway in yeasts. Since sterol C-22 desaturase (ERG5p, CYP61) metabolizes ergosta-5,7,24(28)-trienol into ergosta 5,7,22,24(28)-tetraenol, which is the penultimate compound in the ergosterol biosynthesis, we examined the gene of this enzyme to determine whether this enzyme works in the planthopper YLS. C-22 desaturase genes (ERG5) of YLS of the planthoppers and beetles had four introns in identical positions; such introns are not found in the reported genes of yeasts. Cytochrome P450 cysteine heme-iron ligand signature motif was well conserved among the putative amino acid sequences. The gene expression of the planthopper YLS were strongly suppressed, and the genes possessed nonsense mutations. The accumulation of ergosta 5,7,24(28)-trienol in the planthopper YLS was attributed to the inability of the planthopper YLS to produce functional ERG5p. PMID- 12770583 TI - Sipaucins A-C, sesquiterpenoids from Siparuna pauciflora. AB - The phytochemical investigation of the leaves of Siparuna pauciflora yielded three novel sesquiterpenoids: the germacrane sipaucin A, the elemane sipaucin B and sipaucin C, comprising a new type of carbon skeleton. In addition, four known aporphine alkaloids-nor-boldine, boldine, laurotetanine, and N-methyl laurotetanine-were obtained. The evaluation of the antiplasmodial activity of the isolated compounds against two strains of Plasmodium falciparum (PoW, Dd2) showed a moderate activity of nor-boldine. PMID- 12770584 TI - Charminarone, a seco-pseudoguaianolide from Parthenium hysterophorus. AB - A novel sesquiterpenoid, charminarone, the first seco-pseudoguaianolide, has been isolated along with several known compounds from the whole plant of Parthenium hysterophorus. The structure of the new compound has been settled as 1,10-seco dihydroisoparthenin 1,10-dione from its spectral data as well as from its conversion by reductive coupling reaction with Zn/TiCl(4) reagent to the known compound, anhydrodihydroisoparthenin. The reagent Zn/TiCl(4) has been used here for the first time to construct the seven membered ring of a pseudoguaianolide sesquiterpenoid. PMID- 12770585 TI - Withanolides from Withania coagulans. AB - Two withanolides, 20beta-hydroxy-1-oxo-(22R)-witha-2,5,24-trienolide (1) and withacoagulin (2), along with a known withanolide, 17beta-hydroxy-14alpha, 20alpha-epoxy-1-oxo-(22R)-witha-3,5,24-trienolide (3) were isolated from Withania coagulans. Their structures were elucidated with the help of different spectroscopic techniques. PMID- 12770586 TI - Studies of ent-kaurane diterpenes from Oyedaea verbesinoides for their inhibitory activity on vascular smooth muscle contraction. AB - From the aerial parts of Oyedaea verbesinoides nine ent-kauranes and a sesquiterpene were isolated. ent-9alpha-Hydroxy-kaur-16-en-19-oic acid, ent 15beta-tigloyloxy-9alpha-hydroxy-kaur-16-en-19-oic acid, ent-15beta-angeloyloxy 9alpha-hydroxy-kaur-16-en-19-oic acid, ent-16alpha-hydroxykaurane and 1alpha angeloyloxy-carotol are new for the genus or the species and ent-15beta angeloyloxy-7alpha,9alpha-dihydroxy-kaur-16-en-19-oic acid is reported for the first time. Structure elucidation was based on one and two dimensional NMR as well as ESI and CI-MS analysis. Some diterpenes were proven to exhibit inhibitory effects on smooth muscle contraction on rat aorta. PMID- 12770587 TI - Iridoid glucosides from Randia spinosa (Rubiaceae). AB - An iridoid glucoside: randinoside, along with five known iridoids: galioside, deacetylasperulosidic acid methyl ester, scandoside methyl ester, geniposide and gardenoside, were isolated from the stems of Randia spinosa. The structures were determined by spectroscopic analysis, including 2D NMR techniques. PMID- 12770588 TI - Triterpenoid saponins from Schefflera arboricola. AB - Nine triterpenoid saponins were isolated from the leaves and stems of Schefflera arboricola. The saponins were characterised, on the basis of chemical and spectral evidence, as 3-O-[alpha-L-rhamnopyranosyl-(1-->4)-beta-D glucuronopyranosyl] oleanolic acid, 3-O-[alpha-L-rhamnopyranosyl-(1-->4)-beta-D glucuronopyranosyl] echinocystic acid, 3-O-[beta-D-apiofuranosyl-(1-->4)-beta-D glucuronopyranosyl] oleanolic acid 28-O-beta-D-glucopyranosyl ester, 3-O-alpha-L ramnopyranosyl-(1-->4)-[alpha-L-arabinopyranosyl-(1-->2)-] beta-D glucuronopyranosyl oleanolic acid, 3-O-alpha-L-rhamnopyranosyl-(1-->4)-[alpha-L arabinopyranosyl-(1-->2)-] beta-D-glucuronopyranosyl oleanolic acid 28-O-beta-D glucopyranosyl ester, 3-O-alpha-L-rhamnopyranosyl-(1-->4)-[beta-D galactopyranosyl-(1-->2)-] beta-D-glucuronopyranosyl oleanolic acid, 3-O-alpha-L rhamnopyranosyl-(1-->4)-[beta-D-galactopyranosyl-(1-->2)-] beta-D glucuronopyranosyl oleanolic acid 28-O-beta-D-glucopyranosyl ester, 3-O-beta-D apiofuranosyl-(1-->4)-[alpha-L-arabinopyranosyl-(1-->2)-] beta-D glucuronopyranosyl oleanolic acid and 3-O-beta-D-apiofuranosyl-(1-->4)-[alpha-L arabinopyranosyl-(1-->2)-] beta-D-glucuronopyranosyl oleanolic acid 28-O-beta-D glucopyranosyl ester. PMID- 12770589 TI - The absolute stereochemistry of a diterpene from Ballota aucheri. AB - The semi-synthetic transformation of hispanolone, isolated from Ballota africana, into 6beta-hydroxy-15,16-epoxylabda-8,13(16),14-trien-7-one has established an ent-labdane absolute stereochemistry for a diterpene metabolite originally isolated from B. aucheri. PMID- 12770590 TI - 4alpha-methyl-24beta-ethyl-5alpha-cholesta-14,25-dien-3beta-ol and 24beta ethylcholesta-5, 9(11), 22E-trien-3beta-ol, sterols from Clerodendrum inerme. AB - From the aerial parts of Clerodendrum inerme, two new sterols (4alpha-methyl 24beta-ethyl-5alpha-cholesta-14, 25-dien-3beta-ol and 24beta-ethylcholesta-5, 9(11), 22E-trien-3beta-ol) and a new aliphatic ketone (11-pentacosanone) were isolated together with another known aliphatic ketone (6-nonacosanone) and a diterpene (clerodermic acid). The structure elucidations were based on analyses of physical and spectroscopic data. PMID- 12770591 TI - Pentacyclic triterpenes from Euphorbia stygiana. AB - Two pentacyclic triterpenes, D-friedomadeir-14-en-3beta-yl acetate and D:C friedomadeir-7-en-3beta-yl acetate, named madeiranyl acetate and isomadeiranyl acetate, respectively, were isolated from leaves of Euphorbia stygiana, together with the two known madeiranes, D-friedomadeir-14-en-3-one and D:C-friedomadeir-7 en-3-one, which were obtained from the stem bark. In addition, four known lupane and taraxerane-type triterpenes, namely lupenyl acetate, lupenone, taraxeryl acetate and taraxerone, were also isolated from the same source. Structures were elucidated by physical, chemical and spectroscopic methods (1H NMR, 13C NMR, IR and mass spectra) and by comparison with literature data, and in the case of D:C friedomadeir-7-en-3beta-yl acetate by X-ray analysis as well. PMID- 12770592 TI - Two biflavonoids from Ouratea flava stem bark. AB - In a chemical investigation on the stem bark of Ouratea flava, two biflavonoids: 1-[3-(2,4-dihydroxy-benzoyl)-4,5,6-trihydroxy-2-(4-hydroxy-phenyl)-benzofuran-7 yl] -3-(4-hydroxy-phenyl) -propenone (flavumone A) and 3-(2,4-dihydroxy-benzoyl) 4-hydroxy-2,7-bis-(4-hydroxy-phenyl) -7,8- dihydro-furo[2,3-f]chromen-9-on (flavumone B) were isolated along with five known flavonoids. Their structures were established by various analyses including 2D-NMR spectroscopy. PMID- 12770593 TI - Unique phenolic carboxylic acids from Sanguisorba minor. AB - The unique phenolic carboxylic acids, 4,8-dimethoxy-7-hydroxy-2-oxo-2H-1 benzopyran-5,6-dicarboxylic acid and 2-(4-carboxy-3-methoxystyryl)-2 methoxysuccinic acid were isolated and identified from the whole Sanguisorba minor plant. The known phenolics, gallic acid; ellagic acid; quercetin-3-O-(6" galloylglucose); beta-glucogallin; 2,3-hexahydroxydiphenoyl-(alpha/beta)-glucose; 1-galloyl-2,3-hexahydroxydiphenoyl-alpha-glucose together with its beta-isomer were also characterized. Structures were established by conventional methods of analysis and confirmed by NMR and ESI-MS spectral analysis. PMID- 12770594 TI - Evariquinone, isoemericellin, and stromemycin from a sponge derived strain of the fungus Emericella variecolor. AB - From a strain of the fungus Emericella variecolor derived from the marine sponge Haliclona valliculata, two new natural products, evariquinone and isoemericellin, were isolated after HPLC-UV, -MS, and -NMR studies of the extract and their structures were elucidated by mass spectrometry and NMR experiments. Evariquinone showed antiproliferative activity towards KB and NCI-H460 cells at a concentration of 3.16 microg/ml. Furthermore, the fungus was found to produce the known metabolites stromemycin, shamixanthone, and 7-hydroxyemodin. Chemical degradation, NMR decoupling experiments, and spin-system simulation provided evidence for the double bonds in stromemycin to be all E-configured. ROESY experiments established the monosaccharide moiety to be glucose. PMID- 12770595 TI - Two prenylated flavonoids from the stem bark of Erythrina burttii. AB - From the stem bark of Erythrina burttii, a new isoflavone, 5,2',4'-trihydroxy-7 methoxy-6-(3-methylbut-2-enyl)isoflavone (trivial name, 7-O-methylluteone) and a new flavanone, 5,7-dihydroxy-4'-methoxy-3'-(3-methylbutadienyl)-5'-(3-methylbut-2 enyl)flavanone (trivial name, burttinonedehydrate) along with three known isoflavonoids (8-prenylluteone, 3-O-methylcalopocarpin and genistein) were isolated. The structures were determined on the basis of spectroscopic evidence. PMID- 12770596 TI - Flavone C-glycosides from Lupinus hartwegii. AB - From the aerial parts of Lupinus hartwegii, two new flavone C-glycosides apigenin 7-O-beta-apiofuranosyl-6, 8-di-C-beta-glucopyranoside (1) and apigenin-7-O-beta apiofuranosyl-6-C-beta-glucopyranosyl-8-C-(6z.qprime;-O-E-feruloyl)- beta glucopyranoside (2) have been isolated together with two known isoflavonoid glucosides genistein-7-O-beta-glucopyranoside (3) and genistein-7, 4'-di-O-beta glucopyranoside (4) as well as two known compounds ferulic acid 4-O-beta glucopyranoside (5) and sparteine (6). The structures of the isolated compounds were verified by means of MS and NMR spectral analyses. PMID- 12770597 TI - Phenolic constituents from Drypetes armoracia. AB - The methanol extract of the dried stem bark of Drypetes armoracia Pax & Hoffm. afforded two compounds named drypearmoracein A, (E)-4,5,6,7-tetrahydroxy-2 benzylhept-2-enoic acid and drypearmoracein B, 2,3-dihydroxy-9,10 tetrahydroanthra-1,4-quinone along with five known compounds: friedelan-3 beta ol, friedelin, friedelane-3,7-dione, drypemolundein B and beta-stigmasterol. Their structures were established on the basis of spectroscopic analysis and chemical evidence. PMID- 12770598 TI - Flavonoids from Andrographis lineata. AB - Three flavonoids, 5,7,2',3',4'-pentamethoxyflavone (1), 2'-hydroxy-2,4',6'-tri methoxychalcone (2) and dihydroskullcapflavone I (3), together with 17,19,20 trihydroxy-5beta, 8alpha H, 9beta H,10alpha-labd-13-en-16,15-olactone (4), a known diterpenoid and six known flavonoids, 5-hydroxy-7,8-dimethoxyflavanone (5), 5-hydroxy-7,8,2',3',4'-pentamethoxyflavone (6), 5,2'-dihydroxy-7-methoxyflavanone (7), 5,2'-dihydroxy-7,8-dimethoxyflavone (8), 5,2'-dihydroxy-7-methoxyflavone (9) and 5,2'-dihydroxy-7-methoxyflavone 2'-O-beta-D-glucopyranoside (10) were isolated from the whole plant of Andrographis lineata. The structures of these compounds were elucidated on the basis of spectral and chemical studies. PMID- 12770599 TI - A flavonol tetraglycoside from Sophora japonica seeds. AB - A flavonol tetraglycoside, kaempferol 3-O-alpha-L-rhamnopyranosyl(1-->6)-beta-D glucopyranosyl(1-->2)- beta-D-glucopyranoside-7-O-alpha-L rhamnopyranoside, together with nine known compounds were isolated from the seeds of Sophora japonica L. Their structures were elucidated on the basis of spectral and chemical evidence. PMID- 12770600 TI - Xanthones from the bark of Garcinia merguensis. AB - The bark of Garcinia merguensis yielded 10 xanthones, merguenone, 1,5-dihydroxy 6'-methyl-6'-(4-methyl-3-pentenyl)-pyrano(2',3':3,2)-xanthone, subelliptenone H, 8-deoxygartanin, rheediaxanthone A, morusignin G, 6-deoxyjacareubin, 1,3,5 trihydroxy-4,8-di(3-methylbut-2-enyl)-xanthone, rheediachromenoxanthone and 6 deoxyisojacareubin. The structure of merguenone was determined using spectroscopic techniques, mainly 1D and 2D NMR spectroscopy. PMID- 12770601 TI - Prenylated isoflavonoids from Millettia pervilleana. AB - From the root bark of Millettia pervilleana, which had shown significant cytotoxic activity, a 3-phenylcoumarin, named pervilleanine, two new pterocarpans, pervilline and pervillinine, and one known, emoroidocarpan, were isolated in addition to rotenone and 3alpha-hydroxyrotenone. The anticancer activity of two previously isolated isoflavanones, pervilleanone and 3'-O demethylpervilleanone is reported. PMID- 12770602 TI - Alkaloids from the seeds of Sterculia lychnophora (Pangdahai). AB - Two alkaloids, named sterculinine I and sterculinine II, together with thirteen known compounds were isolated from the ethanol extract of a well-known Chinese traditional drug, Pangdahai (the seeds of Sterculia lychnophora Hance). Their structures were elucidated by NMR, UV, IR and MS spectroscopic analysis. PMID- 12770604 TI - Two acyl sucroses from Petunia nyctaginiflora. AB - Two new acyl sucroses were isolated from the epigeal parts of Petunia nyctaginiflora Juss. (Solanaceae). Their structures were determined to be 2, 3, 4 tri (5-methylhexanoyl)-alpha-D-glucopyranosyl-beta-D-fructofuranoside (2) and 2, 3, 4-tri (6-methylheptanoyl)-alpha-D-glucopyranosyl-beta-D-fructofuranoside (4) on the basis of chemical and spectroscopic evidence. PMID- 12770603 TI - Sesquiterpene lactone glucosides and alkyl glycosides from the fruit of cumin. AB - From the polar portion of the methanolic extract of cumin (fruit of Cuminum cyminum L.), two sesquiterpenoid glucosides, cuminoside A and B, and two alkyl glycosides were isolated together with five known compounds. Their structures were established as (1S,5S,6S,10S)-10-hydroxyguaia-3,7(11)-dien-12,6-olide beta-D glucopyranoside, (1R,5R,6S,7S,9S,10R,11R)-1,9-dihydroxyeudesm-3-en-12,6-olide 9-O beta-D-glucopyranoside, methyl beta-D-apiofuranosyl-(1-->6)-beta-D glucopyranoside and ethane-1,2-diol 1-O-beta-D-apiofuranosyl-(1-->6)-beta-D glucopyranoside, respectively. PMID- 12770605 TI - Phenylethanoid and aliphatic alcohol glycosides from Acanthus ilicifolius. AB - A phenylethanoid glycoside (ilicifolioside A) and an aliphatic alcohol glycoside (ilicifolioside B), have been isolated from the aerial parts of Acanthus ilicifolius, together with eight known compounds. Their structures were determined from spectroscopic analyses. PMID- 12770606 TI - Flavonol and drimane-type sesquiterpene glycosides of Warburgia stuhlmannii leaves. AB - An investigation of methanolic extract of Warburgia stuhlmannii leaves has led to the isolation of two new drimane-type sesquiterpene glycosides characterized as mukaadial 6-O-beta-D-glucopyranoside, mukaadial 6-O-alpha-L-rhamnopyranoside together with two other novel flavonol glycosides identified as 3',5'-O dimethylmyricetin 3-O-beta-D-2",3"-diacetylglucopyranoside and 3'-O methylquercetin 3-O-beta-D-2",3",4"-triacetylglucopyranoside. The known compounds; mukaadial, deacetylugandensolide, quercetin, kaempferol, kaempferol 3 O-alpha-L-rhamnopyranoside, quercetin 3-O-beta-D-glucopyranoside, kaempferol 7-O beta-D-glucopyranoside, myricetin 3-O-alpha-L-rhamnopyranoside, quercetin 3-O alpha-L-rhamnopyranoside, quercetin 3-O-sophoroside and isorhamnetin 3-O-beta-D glucopyranoside were also isolated from the same extract. PMID- 12770608 TI - Contribution of salt intake to insulin resistance associated with hypertension. AB - It is well-known that excessive salt intake induces elevation of blood pressure and that hypertension often coexists with insulin resistance. However, the contribution made by salt intake to the development of insulin resistance remains unclear. In this minireview, the insulin resistance seen in three salt-sensitive rat models (the high-salt-fed Sprague-Dawley rat, the Dahl salt-sensitive rat and the chronically angiotensin II (AII)-infused rat) are presented. One notable observation common to all three models was that, despite their insulin resistance, the early insulin signaling steps leading from activation of IR and IRSs to activation of PI 3-kinase Akt were apparently enhanced. This suggests that the molecular mechanism underlying the insulin resistance related to the salt-sensitive hypertension is unique. Other factors known to cause insulin resistance--e.g., obesity--actually suppress early insulin signaling, but for insulin resistance related to high-salt intake, the impaired step must be further downstream in the insulin signaling pathway. What's more, increased oxidative stress appears to be crucially involved in both AII- and high-salt-induced insulin resistance. Additional study will be necessary to fully clarify the mechanism underlying insulin resistance induced by a high-salt diet; nonetheless, the findings presented suggest the importance of developing new therapeutic approaches--e.g., potassium supplementation and anti-oxidant administration--to the treatment of insulin resistance and hypertension. PMID- 12770609 TI - Hormonal status modifies renin-angiotensin system-regulating aminopeptidases and vasopressin-degrading activity in the hypothalamus-pituitary-adrenal axis of male mice. AB - Local renin-angiotensin systems (RAS) have been postulated in brain, pituitary and adrenal glands. These local RAS have been implicated, respectively, in the central regulation of the cardiovascular system and body water balance, the secretion of pituitary hormones and the secretion of aldosterone by adrenal glands. By other hand, it is known that the hypothalamus-pituitary-adrenal (HPA) axis is involved in blood pressure regulation, and is affected by sex hormones. The aim of the present work is to analyze the influence of testosterone on RAS regulating aminopeptidase A, B and M activities and vasopressin-degrading activity in the HPA axis, measuring these activities in their soluble and membrane-bound forms in the hypothalamus, pituitary and adrenal glands of orchidectomized males and orchidectomized males treated subcutaneously with several doses of testosterone. The present data suggest that in male mice, testosterone influences the RAS- and vasopressin-degrading activities at all levels of the HPA axis. PMID- 12770610 TI - A multicenter, randomized, placebo controlled, multiple-dose, safety and pharmacokinetic study of AIT-082 (Neotrofin) in mild Alzheimer's disease patients. AB - A phase 1, randomized, double-blind, placebo-controlled, dose escalation study of the purine derivative, AIT-082 (Neotrofin, NeoTherapeutics) was conducted in mild Alzheimer's disease (AD) patients to evaluate multiple-dose safety, tolerability, and pharmacokinetics. Possible short-term effects of AIT-082 on cognition and memory were preliminarily investigated. AIT-082 is currently being developed as a potential treatment for Alzheimer's disease and other neurological disorders. Pre clinical studies indicate that AIT-082 has memory enhancing properties, stimulates neuritogenesis and the production of neurotrophic factors. Patients received an oral dose of AIT-082 or placebo daily for one week. Thirty-six AD patients were divided into three dose cohorts; each dose cohort consisted of twelve patients with 8 patients randomized to AIT-082 and 4 to placebo. The 3 doses of AIT-082 evaluated in this study were 100 mg/day, 500 mg/day, and 2,000 mg/day. There were no serious adverse events at any dose and the drug was well tolerated without significant side effects. AIT-082 was orally and rapidly absorbed, resulting in peak serum concentrations within 2 hours with an elimination half-life of approximately 20 hours. Higher doses resulted in corresponding increases in peak concentrations and areas under the curve (AUC). There was an approximate 2-fold accumulation in AIT-082 with daily dosing (as reflected by the AUC) at steady state. There were no significant differences by treatment arm on the clinical or neuropsychological evaluations. AIT-082 was rapidly absorbed by the oral route with a half-life suitable for dosing once or twice daily. No problems with tolerability or safety were found. AIT-082 appears suitable for testing in larger clinical trials for the treatment of AD and other neurologic disorders. PMID- 12770611 TI - Antibody to beta-amyloid protein increases acetylcholine in the hippocampus of 12 month SAMP8 male mice. AB - Amyloid beta protein (Abeta) is the primary constituent of plaque seen in Alzheimer's disease. Abeta is proposed to play an etiological role in Alzheimer's disease and to be a cause of the decrease in the level of acetylcholine in the hippocampus. The SAMP8 strain of mouse develops age-related increases in Abeta and deficits in learning and memory by 12 months of age. We examined in 12 month old SAMP8 mice the effects of giving antibody to Abeta by septal or intracerebroventricular (ICV) injection on acetylcholine levels in the hippocampus. Antibody to Abeta increased acetylcholine in the hippocampus over 100% after ICV injection and over 200% after septal injection. Injection of rabbit serum, antibody directed towards mouse IgG, or a blocking antibody directed towards human interleukin-1beta were without effect. These results suggest that antagonism of Abeta increases acetylcholine concentrations in the hippocampus, an area important for learning and memory. PMID- 12770613 TI - MPTP-induced reactive oxygen species promote cell death through a gradual activation of caspase-3 without expression of GRP78/Bip as a preventive measure against ER stress in PC12 cells. AB - Glucose-regulated protein 78 (GRP78)/Immunoglobulin binding protein (Bip) is a chaperone which functions to protect cells from endoplasmic reticulum (ER) stress. GRP78/Bip is expressed following ER stress induced by thapsigargin, tunicamycin or chemical factors. However, the mechanism of progression of ER stress against stress factors is still obscure. We examined whether reactive oxygen species (ROS) were involved in GRP78/Bip expression and caspase-3 activity was induced in PC12 cells using 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) to produce ROS. We report that PC12 cells lost viability in the presence of MPTP for 24 hours as a partial effect of ROS. We also show that N-acetyl-L cysteine diminished the MPTP-induced apoptosis with expunction of ROS. Furthermore, we observed that GRP78/Bip was not up-regulated and the caspase-3 activity was increased in the presence of MPTP. These results suggest that insubstantial ROS do not contribute to the ER stress-mediated cell death while caspase-3 is involved in ROS-promoted cell death in MPTP-treated cells. PMID- 12770612 TI - Synergistic effect of angiopoietin-1 and vascular endothelial growth factor on neoangiogenesis in hypercholesterolemic rabbit model with acute hindlimb ischemia. AB - Vascular endothelial growth factor (VEGF) and angiopoietin-1 (Ang1) are essential for vascular integrity and development. The purpose of the study was to test the hypothesis that Ang1 will promote angiogenic response to VEGF in the spontaneous Watanabe heritable hypercholesterolemic (WHHL) rabbit model of acute hindlimb ischemia. Immediately after the ligation of the external iliac artery and the excision of the common and superficial femoral artery in one female WHHL rabbit, 250 microg of phVEGF(165) (n = 8), 500 microg of pAng1* (n = 8), or 250 microg of phVEGF(165) plus 500 microg of pAng1* (n = 8) was injected intramuscularly into the ischemic hindlimb muscles. Gross appearance of ischemic limb, collateral vessel formation and limb perfusion were assessed 30 days after treatment. The incidence of ischemic limb necrosis was higher in the animals treated by phVEGF(165) or by pAng1* than in those treated by phVEGF(165) plus pAng1* (100%, 75% and 14.3%, respectively; P = 0.002). Animals in the combination therapy group had a significantly higher calf blood pressure ratio at day 30 (VEGF plus Ang1* = 0.84 +/- 0.06; VEGF = 0.54 +/- 0.01; Ang1* = 0.59 +/- 0.05; P < 0.01). A combination therapy of VEGF plus Ang*1 had a significantly higher (P < 0.01) angiographic score than either therapy alone. Capillary density (P < 0.05) and capillary/muscle fiber ratio (P < 0.01) of the combination therapy group were also significantly higher than that of either therapy alone. In conclusion, Ang1 can potentiate the angiogenic response to VEGF in the hyperlipidemic rabbit model of acute hindlimb ischemia. Intramuscular administration of cytokines on revascularization of the ischemic hindlimb model of hyperlipidemic rabbit is feasible. PMID- 12770614 TI - The regulation of inducible nitric oxide synthase gene expression induced by lipopolysaccharide and tumor necrosis factor-alpha in C6 cells: involvement of AP 1 and NFkappaB. AB - The roles of AP-1 and NFkappaB in the regulation of inducible nitric oxide synthase (iNOS) mRNA expression induced by the combination of lipopolysaccharide and tumor necrosis factor-alpha (LT) in C6 cells were examined in the present study. The iNOS mRNA level and NO release were increased by several cytokines alone or combination treatments at 24 hr. LT-induced iNOS mRNA level was maximally increased at 6 hr and maintained at higher level at least up to 24 hr. At 6 hr, iNOS protein level and NO release were also increased by LT. By western blot analysis, AP-1, such as Fra-1, Jun B, and phospho-CREB protein levels were increased by LT and translocation of NFkappaB p52 from the cytoplasm to the nucleus was increased. In addition, phosphorylations of MAPKs (ERK 1/2, p38, JNK 1/2) were increased by LT. LT-induced iNOS mRNA level was inhibited by PD98059 (MEK 1/2 inhibitor), SB203580 (p38 inhibitor), and cycloheximide (a protein synthesis blocker), indicating that the phosphorylation of ERK 1/2 and p38, and on-going protein synthesis are necessary for LT-induced iNOS expression. Electrophoretic mobility shift assay (EMSA) showed that AP-1 and NFkappaB DNA binding activities were increased at 6 hr and these AP-1 and NFkappaB DNA bands increased by LT were super-shifted when Fra-1, Jun B, or NFkappaB p50 antibody was coincubated. These findings strongly suggest that, in C6 cells, Fra-1, Jun B, NFkappaB p50, and NFkappaB p52 appear to be involved in the regulation of iNOS mRNA induced by LT. PMID- 12770615 TI - Petrosaspongiolide M reduces morphine withdrawal in vitro. AB - The bee venom phospholipase A(2) (PLA(2)) inhibitory activity of petrosaspongiolide M (PM), a marine metabolite displaying a potent anti inflammatory activity and able to covalently bind and block group II and III secretory PLA(2) enzymes, has been investigated by mass spectrometry and molecular modeling. The model reveals interesting insight on the PM-PLA(2) inhibition process and may prove useful in the design of new anti-inflammatory agents targeting PLA(2) secretory enzymes. In this paper, the effect of PM has been investigated on opiate withdrawal in an in vitro model. After a 4 min in vitro exposure to morphine a strong contracture of guinea pig isolated ileum was observed after the addition of naloxone. PM treatment 1 x 10(-8), 5 x 10(-8), 1 x 10(-7) M was able to reduce morphine withdrawal. These results suggest that PM effect in this in vitro model of opiate withdrawal may be due to extracellular type II PLA(2) inhibition. PMID- 12770616 TI - Effects of dopamine agonists bromocriptine, pergolide, cabergoline, and SKF-38393 on GDNF, NGF, and BDNF synthesis in cultured mouse astrocytes. AB - We examined the stimulatory effects of the dopamine agonists bromocriptine, pergolide, cabergoline, and SKF-38393 on the synthesis and secretion of neurotrophic factors (nerve growth factor, NGF; brain-derived neurotrophic factor, BDNF; and glial cell line-derived neurotrophic factor, GDNF) in cultured mouse astrocytes, and clarified the role of dopamine D1 and D2 receptors in these effects. Bromocriptine, a D2 agonist, elevated NGF levels in the culture medium 6.8-fold vs. control, and significantly decreased GDNF and BDNF levels, at 24 h. Both pergolide, a D1/D2 agonist, and cabergoline, a D2/weak D1 agonist, rapidly elevated NGF and GDNF levels at 4-6 h, respectively to 21- and 1.5-fold, respectively, and 84- and 9-fold, respectively, of control levels at 24 h. SKF 38393, a D1 agonist, elevated NGF and GDNF levels to 20- and 2.8-fold of controls, respectively, at 24 h. Relative levels of NGF and GDNF mRNA detected by Northern blot analysis or semiquantitative reverse transcriptase-polymerase chain reaction confirmed that increases in levels of the 2 proteins in culture medium were due to overexpression as opposed to leakage from cells. Cabergoline rapidly increased GDNF mRNA expression at 4 h, producing a potent and long-lasting increase in GDNF levels. Bromocriptine significantly suppressed GDNF synthesis. These findings suggest that stimulation of dopamine D1 receptors may be required for GDNF synthesis and secretion, and that concurrent stimulation of dopamine D1 and D2 receptors may augment synthesis and secretion of NGF and GDNF. These dopamine agonists may play a role in neuronal survival by stimulating NGF and GDNF synthesis in the brain, and as drugs are good candidates as NGF and GDNF inducers. PMID- 12770619 TI - Virus or not? Phylogenetics of polydnaviruses and their wasp carriers. AB - Our current, still limited, understanding of the comparative biology and evolution of polydnaviruses (PDVs) is reviewed, especially in the context of the possible origins of these parasitoid viruses and of their coevolution with carrier wasps. A hypothetical scenario of evolution of PDVs from ascovirus (or ascovirus-like) ancestors is presented, with examples of apparent extant transitional forms. PDVs appear, in the case of bracoviruses, to show phylogenetic relationships that mirror those of their wasp carriers: with ichnoviruses, the picture is less clear. Ongoing sequencing studies of entire PDV genomes from diverse wasp species are likely to greatly contribute to our understanding of PDV evolution. PMID- 12770617 TI - Step by step removal of hyperforin and hypericin: activity profile of different Hypericum preparations in behavioral models. AB - Herbal extracts of Hypericum perforatum L. (St. John's wort, SJW) are now successfully competing for status as a standard antidepressant therapy. Because of this, great effort has been devoted to identifying the antidepressive active compounds. In the present study we used the following strategy to evaluate the relative pharmacological importance of various extract components: 1. preparation of an hydroalcoholic SJW extract containing both hyperforin (3.2%) and hypericin (0.15%) (extract A); 2. step by step removal of hyperforin and hypericin led to the following extracts: Extract B, devoid of hyperforin but still containing hypericin (0.14%) and Extract C, free of hypericin and hyperforin but enriched in flavonoids ( approximately 12%). We characterized the in vivo activity profile of all three preparations using the tail suspension test (TST) in mice and the forced swimming test (FST) in rats as screening models. We further investigated the activity of pure hyperforin. Extract B and C (500 mg/kg each) as well as pure hyperforin (8 mg/kg) significantly shortened immobility time in the TST after acute pre-treatment whereas extract A was inactive. In the FST all three extracts decreased immobility time in a dosage of 500 mg/kg after acute as well as after repeated treatment. The present results clearly show that an SJW extract free of hyperforin and hypericin exerts antidepressant activity in behavioral models, supporting our working hypothesis that flavonoids are part of the constituents responsible for the therapeutic efficacy of SJW extracts. We also could show that hyperforin contributes to the beneficial properties of SJW extract, confirming the hypothesis that the crude SJW extract contains several constituents with antidepressant activity. PMID- 12770620 TI - Polydnavirus genome: integrated vs. free virus. AB - Polydnaviruses are unique because of their obligatory association with thousands of parasitoid wasp species from the braconid and ichneumonid families of hymenopterans. PDVs are injected into the parasitized hosts and are essential for parasitism success. However, polydnaviruses are also unique because of their genome composed of multiple dsDNA segments. Cytological evidence has recently confirmed the results of genetic and molecular analyses indicating that PDV segments were integrated in the wasp genome. Moreover a phylogenetic study performed using the age of available fossils to calibrate the molecular clock indicated that the polydnaviruses harboured by braconid wasps have resided within the wasp genome for approximately 70 million years. In the absence of horizontal transmission, the evolution of the PDV genomes has been driven exclusively by the reproductive success they have offered the wasps. The consequences of this particular selection pressure can be observed in the gene content of certain PDV genomes from which increasing sequence data are available. Molecular mechanisms already identified could be involved in the acquisition and loss of genes by the PDV genomes and lead us to speculate on the definition of the virus genome. PMID- 12770621 TI - Origin and evolution of polydnaviruses by symbiogenesis of insect DNA viruses in endoparasitic wasps. AB - During oviposition, many endoparasitic wasps inject virus-like particles into their insect hosts that enable these parasitoids to evade or directly suppress their hosts' immune system, especially encapsulation by hemocytes. These particles are defined as virions that belong to viruses of the two genera that comprise the family Polydnaviridae, bracoviruses (genus Bracovirus) transmitted by braconid wasps, and ichnoviruses (genus Ichnovirus) transmitted by ichneumonid wasps. Structurally, bracovirus virions resemble nudivirus and baculovirus virions (family Baculoviridae), and ichnovirus virions resemble those of ascoviruses (family Ascoviridae). Whereas nudiviruses, baculoviruses and ascoviruses replicate their DNA and produce progeny virions, polydnavirus DNA is integrated into and replicated from the wasp genome, which also directs virion synthesis. The structural similarity of polydnavirus virions to those of viruses that attack the wasps' lepidopteran hosts, along with polydnavirus transmission and replication biology, suggest that these viruses evolved from insect DNA viruses by symbiogenesis, the same process by which mitochondia and chloroplasts evolved from bacteria. Molecular evidence supporting this hypothesis comes from similarities among structural proteins of ascoviruses and the Campoletis sonorensis ichnovirus. Implications of this hypothesis are that polydnaviruses evolved from viruses, but are no longer viruses, and that DNA packaged into polydnavirus virions is not viral genomic DNA per se, but rather wasp genomic DNA consisting primarily of wasp genes and non-coding DNA. Thus, we suggest that a better understanding of polydnaviruses would result by viewing these not as viruses, but rather as a wasp organelle system that evolved to shuttle wasp genes and proteins into hosts to evade and suppress their immune response. PMID- 12770622 TI - Expression of Cotesia kariyai polydnavirus genes in lepidopteran hemocytes and Sf9 cells. AB - The parasitic wasp Cotesia kariyai carries polydnavirus (CkPDV) which is an indispensable factor for the successful parasitization by the wasp. One of its surface proteins has been identified as an immunoevasive mediator from the cellular defense reactions of the host armyworm Pseudaletia separata, thereby it was named immunoevasive protein (IEP). In the present study, we demonstrated that anti-IEP antibody did not suppress the CkPDV infection of Sf9 cells but decreased its infection of P.separata hemocytes, thus indicating that IEP is not essential for CkPDV to enter into the target cells but is important for evading from the attack of the hemocytes. Three genes of CkPDV expressed in Sf9 cells were isolated and characterized. Two of them (CkV0.8, CkV0.9) are novel genes but another one (CkV2.0) is the same gene with the one we previously identified in the parasitized armyworm larvae. Although these genes reside in different DNA segments of CkPDV genome, all of them are expressed in the hemocytes of the parasitized armyworm larvae. These gene transcripts are first detected at 2 h after parasitization, and the expressions of CkV0.8 and CkV0.9 were gradually decreased after reaching the maximum level at 4 h after parasitization. However, the expression of CkV2.0 continues to be increased at least for 10 h after parasitization. PMID- 12770623 TI - Two Hyposoter didmator ichnovirus genes expressed in the lepidopteran host encode secreted or membrane-associated serine and threonine rich proteins in segments that may be nested. AB - We present in this work two novel Hyposoter didymator ichnovirus genes expressed in parasitized Spodoptera larvae. These genes, named HdCorfS6 and HdGorfP30, are unrelated and present in two different genome segments, possibly nested, SH-C and SH-G respectively. HdCorfS6 encodes a predicted transmembrane protein, putatively glycosylated. HdCorfS6 transcripts appear to be abundant in lepidopteran host hemocytes compared to the other tissues analyzed. The second gene described, HdGorfP30, is well expressed in hemocytes, but also in other tissues, such as the fat body, nervous system and epidermis. This gene is peculiar since it presents 17 perfectly conserved repeated sequences arranged in tandem arrays. Each of these repeats contains 58% of serine and threonine residues and therefore several potential sites for glycosylation. This mucin-like protein, predicted as highly glycosylated, could be involved in host immune suppression. PMID- 12770624 TI - Polydnavirus integration in lepidopteran host cells in vitro. AB - The long-term persistence of polydnavirus (PDV) DNA in infected lepidopteran cell cultures has suggested that at least some of the virus sequences become integrated permanently into the cell genome. In the current study, we provide supportive evidence of this event. Cloned libraries were prepared from two different Lymantria dispar (gypsy moth) cell lines that had been maintained in continuous culture for more than five years after infection with Glyptapanteles indiensis PDV (GiPDV). Junction clones containing both insect chromosomal and polydnaviral sequences were isolated. Precise integration junction sites were identified by sequence comparison of linear (integrated) and circular forms of the GiPDV genome segment F, from which viral sequences originated. Host chromosomal sequences at the site of integration varied between the two L. dispar cell lines but virus sequence junctions were identical and contained a 4-base pair CATG palindromic repeat. The GiPDV segment F does not encode any self replication or self-insertion proteins, suggesting a host-derived mechanism is responsible for its in vitro integration. The chromosomal site of one junction clone contained sequences indicative of a new L. dispar retrotransposon, including a putative reverse transcriptase and integrase located upstream of the site of viral integration. A potential mechanism is proposed for the integration of PDV DNA in vitro. It remains to be seen if integration of the virus also occurs in the lepidopteran host in vivo. PMID- 12770625 TI - Similar polydnavirus genes of two parasitoids, Cotesia kariyai and Cotesia ruficrus, of the host Pseudaletia separata. AB - Endoparasitoids can achieve successful development in suitable hosts with the help of polydnavirus (PDV), if it is injected into the host with their eggs at parasitization. It was predicted that different species of endoparasitoids that parasitize the same host successfully use a similar system to avoid the defense reaction from the host. In this study, we identified similar PDVs genes. In two endoparasitoids, Cotesia kariyai (Cky) and Cotesia ruficrus (Crf), which can parasitize the same host species, Pseudaletia separata. The electrophoretic profiles of nondigestive segments of PDV in the two parasitoids were different. However, genomic Southern hybridization using reciprocal probes between the two species indicated a similar sequence of PDV genes. Two cDNA libraries were established and screened by a Cky probe. Cky811 was 695-bp, Crf111 was 690-bp, and the predicted peptide sequences were 162aa and 157aa, respectively. The two transcripts had 72.8% similarity with each other and resembled a C-type lectin2 domain. Clones of Cky811 from C. kariyai and Crf111 from C. ruficrus were expressed in hemocytes of the hosts that were injected with PDV plus venom. The Cky811 gene was expressed in hemocytes and fat body of the Pseudaletia host. Hybridization analysis revealed that the Cky811 gene was expressed 6 hrs after injection of Cky-PDV into nonparasitized hosts with venom, and the hybridization signal declined with time. The genetic distance to the lectin of other insects was closer to P. americana than to Lepidopteran insects. PMID- 12770626 TI - Cross-protection experiments with parasitoids in the genus Microplitis (Hymenoptera: Braconidae) suggest a high level of specificity in their associated bracoviruses. AB - The immunological and developmental effects of bracoviruses (BVs) from three parasitoids in the genus Microplitis (Braconidae: Microgastrinae) were compared in the hosts Pseudoplusia includens and Heliothis virescens (Lepidoptera: Noctuidae). Southern blotting experiments indicated that viral DNAs from Microplitis demolitor bracovirus (MdBV) cross-hybridized with viral DNAs from Microplitis croceipes bracovirus (McBV) and Microplitis mediator bracovirus (MmBV) under conditions of high stringency. Injection of calyx fluid plus venom from each parasitoid species dose-dependently delayed development of P. includens and H. virescens. Each virus also inhibited pupation of P. includens but not H. virescens. In situ hybridization experiments indicated that MdBV and McBV persistently infect hemocytes in both hosts while MmBV persistently infects hemocytes in P. includens but not H. virescens. While MdBV infection induced a loss of adhesion by most plasmatocytes, McBV and MmBV infection induced a loss of adhesion in less than 50% of cells. Cross-protection experiments indicated that calyx fluid plus venom from one species usually protected progeny of another species from encapsulation but did not always promote successful development. PMID- 12770627 TI - Overt viral diseases induced from apparent latency following parasitization by the ichneumonid wasp, Hyposoter exiguae. AB - Latent and/or asymptomatic viral infections are commonplace in insects, but factors inducing overt disease are poorly understood. Here we show that in Trichoplusia ni larvae parasitized by the ichneumonid wasp, Hyposoter exiguae, overt, lethal disease may on occasion be observed; however, disease has been consistently absent in control (non-parasitized) larvae. Thus far, we have detected two such diseases, one of which is caused by a granulosis virus affecting primarily fat body tissue. The other is associated with the presence of two viruses replicating together in larval midgut epithelial cells; of these, one has been identified as a non-occluded form of TnCPV. Since H. exiguae carries a polydnavirus, which is delivered to host larvae during oviposition, it is tempting to speculate that viral latency may in some cases be broken through immunosuppressive activity resulting from insect parasitism. PMID- 12770628 TI - Fate of polydnavirus DNA of the egg-larval parasitoid Chelonus inanitus in the host Spodoptera littoralis. AB - In situ hybridizations show that 5 min after parasitization, polydnavirus DNA is in close vicinity of the parasitoid egg, but 5 h later also in the yolk and partially in the host embryo. Fifteen hours after parasitization, the viral DNA is seen all over the host embryo and hardly in the yolk. The tissue distribution of the viral DNA was analysed and quantified by dot blots in the fifth instar parasitized larvae. On a per host basis, haemocytes and fat body contained the highest amount of viral DNA, while nervous tissue, intestinal tract and carcass contained less. Of the three viral segments tested, all were found in all tissues. Relative to the quantity of host DNA, viral DNA was most abundant in haemocytes, about five times less abundant in fat body and nervous tissue and about 25 times less abundant in intestinal tract. The total quantity of viral DNA per host was 444+/-145 pg which is similar to the quantity injected by the wasp; thus, the viral DNA persists throughout parasitization. The parasitoid larva contains 820+/-80 pg viral DNA integrated in the genome. This illustrates that the dose of viral DNA injected in virions represents approximately one third of the total viral genomic information present in a host at a late stage of parasitism. PMID- 12770629 TI - Three related TrIV genes: comparative sequence analysis and expression in host larvae and Cf-124T cells. AB - We report on the cloning and sequencing of two Tranosema rostrale ichnovirus (TrIV) genes, and assess their relatedness to TrV1, the gene encoding the most abundant TrIV transcript in last-instar Choristoneura fumiferana larvae parasitized by T. rostrale. One of the two newly isolated genes, TrV2, features an organization similar to that of TrV1, with one intron flanked by two exons; it encodes a 102 amino acid protein showing 79% similarity to TrV1. The third gene, TrV4, encodes a larger protein (143 aa) displaying similarity to the other two only over the first approximately 50 amino acid residues of its sequence; the remaining portion contains an imperfect octad repeat. Although the TrV4 gene contains only one exon, it has an intron similar in size and sequence to that of TrV1 and TrV2; in fact, the non-coding regions of all three genes show higher sequence identity than the coding regions, pointing to their common origin. Southern analysis suggests that each gene maps to a different TrIV genome segment, with homologous sequences apparently present on other segments. TrV1 and TrV4 transcription in penultimate (5th) instar hosts, parasitized shortly after the molt, was strong for both genes 1 and 2 days p.p., with transcript abundance decreasing after the final molt; thus, neither of these genes is upregulated during induction of developmental arrest in last-instar hosts. Cf-124T cells inoculated with T. rostrale calyx fluid showed significant levels of apoptosis 24 72 h p.i.; TrV1 was detected in the culture medium, suggesting that this and/or other TrIV-encoded proteins may be responsible for the observed cytopathic effect. Southern and Northern analyses, using DNA and RNA extracted from infected Cf-124T cells, revealed the presence of both TrV1- and TrV4-carrying genome segments and transcripts, but neither DNA, at least in episomal form, nor mRNA persisted for more than a few days p.i. This in vitro system may provide a suitable starting point for the study of TrIV gene functions. PMID- 12770630 TI - Active suppression of D. melanogaster immune response by long gland products of the parasitic wasp Leptopilina boulardi. AB - To develop inside their insect hosts, endoparasitoid wasps must either evade or overcome the host's immune system. Several ichneumonid and braconid wasps inject polydnaviruses that display well-studied immune suppressive effects. However, little is known about the strategies of immunoevasion used by other parasitoid families, such as figitid wasps. The present study provides experimental evidence, based on superparasitism and injection experiments, that the figitid species Leptopilina boulardi uses an active mechanism to suppress the Drosophila melanogaster host immune response, i.e. the encapsulation of the parasitoid eggs. The immune suppressive factors are localised in the long gland and reservoir of the female genital tractus, where virus-like particles (VLPs) have been observed. Parasitism experiments using a host tumorous strain indicate that these factors do not destroy host lamellocytes but that they impair the melanisation pathway. Interestingly, they are not susceptible to heating and are not depleted with prolonged oviposition experience, in contrast to observations reported for L. heterotoma, another figitid species. The mechanisms that prevent encapsulation of eggs from L. boulardi and L. heterotoma differ in several respects, suggesting that different physiological strategies of immunosuppression might be used by specialised and generalist parasitoids. PMID- 12770631 TI - Characterization of Campoletis sonorensis ichnovirus unique segment B and excision locus structure. AB - Polydnaviruses (PDVs) are segmented, symbiotic, double-stranded DNA viruses that are vertically transmitted as proviruses within the genomes of some parasitoid Hymenoptera. The PDV associated with the ichneumonid wasp Campoletis sonorensis (CsIV) consists of 24 non-redundant DNA segments varying in size from approximately 6 to 20 kbp. CsIV segment B, one of the smallest genome segments, was sequenced and the excision sites of the proviral segment were characterized. The segment B sequence was 83.2% non-coding with only two open reading frames (ORFs). Some non-coding sequences have similarities to database sequences and were likely pseudogenic, but most were unrelated to known nucleic acid or predicted protein sequences. One ORF, BHv0.9, encodes a member of the rep gene family and was expressed only in parasitized insects while transcription of the other ORF could not be detected. Previously, a third region of the segment was shown to hybridize to 0.6 and 1.2 kb poly A+ RNAs from female wasps during virus replication (Theilmann and Summers, 1988) but this region did not have an identifiable ORF in the determined sequence. In contrast to CsIV segment W, segment B had little repetitive sequence. The segment B proviral integration locus contains a 59 bp direct imperfect repeat. Further analyses of this integration locus demonstrated that segment B was excised from wasp genomic DNA with flanking sequences at the integration site rejoined after segment excision. The segment B "excision locus" retained one of the two copies of the 59 bp repeat sequence with the other repeat present in the excised segment. The data indicate that Ichnovirus segments have distinctive characteristics possibly reflecting functional co-evolution between the wasp and individual types of polydnavirus segments. PMID- 12770632 TI - Persistence and expression of Cotesia congregata polydnavirus in host larvae of the tobacco hornworm, Manduca sexta. AB - The gregarious braconid wasp Cotesia congregata parasitizes host larvae of Manduca sexta, and several other sphingid species. Parasitism induces host immunosuppression due to the disruptive action of the wasp's polydnavirus (PDV) on host blood cells. During the initial stages of parasitism, these cells undergo apoptosis followed by cell clumping, which clears the hemolymph of a large number of cells. In this study, the persistence and expression of Cotesia congregata PDV (CcPDV) were examined using Southern and Northern blots, respectively. Digoxygenin-labelled total polydnaviral DNA was used to probe genomic DNA isolated from fat body and brains of hosts with emerged wasps taken 6 days following egress of the parasitoids, and significant cross-hybridization between the host fat body genomic DNA with viral DNA was seen. Thus, the virus persists in the host for the duration of parasitism, even during the post-emergence period, and may even be integrated in the host caterpillar DNA. Viral gene expression was examined using Northern blots and probes to the Cotesia rubecula CrV1 homolog, and the CrV1-like mRNAs were expressed as early as 4 h post parasitization for at least 72 h and faint hybrization is even seen at the time the wasps eclose. In contrast, in Pieris rapae larvae the CrV1 transcript is expressed only for a brief time, during which time hemocyte function is disrupted. The effect is transitory, and hemocytes regain their normal functions after the parasites emerge as first instars.The genome of CcPDV contains one copy of the CrV1-like homolog as shown on Southern blots of viral genomic DNA. In conjunction with our earlier studies of the PDV-encoded early protein 1, the current work suggests multiple viral transcripts are produced following parasitization of the host, and likely target host hemocytes to induce their apoptosis, thereby preventing encapsulation of the parasitoid's eggs. Whether viral DNAs are integrated in the host's genomic DNA remains to be proven, but our results provide preliminary evidence that viral DNAs are detected in the host's fat body cells examined at the time of wasp emergence and several days later. PMID- 12770633 TI - Local or global asymmetry in gait of people without impairments. AB - Using two consecutive gait cycles, simultaneous and bilateral kinetic gait data, the main objectives of this study were (a) to identify the main functional roles of ankle, knee and hip extensors/flexors, and (b) to determine whether the action taken by these muscle groups appears to be symmetric or not. Gait of our able bodied subjects appears to be asymmetric with significant differences noted between each two corresponding peak muscle moment values. Using principal component analysis (PCA) as a curve structure detection method, task discrepancies were recognized when comparisons were made between each two corresponding representative moment curves at each joint (local asymmetry). Muscle moment behaved symmetrically when the right limb representative curve was compared to its corresponding principal component (PC) at the contralateral limb. Gait of able-bodied subjects appears to be symmetric, while control and propulsion were recognized as two major roles of the extensors and flexors (global gait asymmetry). Symmetrical behavior of the lower limbs should be considered a consequence of local asymmetry which indicates different levels of within and between muscle activities developed at each joint during gait cycles. PMID- 12770634 TI - A proposed test to support the clinical movement analysis laboratory accreditation process. AB - This paper describes a testing methodology and resultant set of four variables that can be used to quickly and easily document the correct installation, configuration, and combined working status of force platform (FP) and three dimensional (3D) motion capture components of a clinical movement analysis (CMA) laboratory. Using a rigid, rod-shaped testing device, CMA laboratory data are collected simultaneously from the FP and motion capture components (typically, video-based kinematic measurements) as the device is manually loaded while being pivoted broadly about a point on the FP. Using a computational method based on static equilibrium, it is possible to independently measure the rod's orientation and tip position during the moving trial, using FP derived data exclusively, and to compare these estimates to rod orientation and tip position estimates derived exclusively from the motion capture component. The motion laboratory accreditation test (MLAT) variables include: the difference (angle) between the orientation of the long axis of the testing device as independently determined from kinematic measures (motion capture component) and the FP derived data; and the difference (x, y, z) between the center of pressure position (FP derived) and the position of the testing device tip (motion capture derived) that loads the FP. A numerical dynamics model was explored to evaluate the appropriateness of the static equilibrium-based FP data model and to determine guidelines for testing device movement frequency and FP loading. The MLAT technique provides a simple means of detecting the combined presence of errors from many sources, several of which are explored in this paper. The MLAT has been developed to help meet one criteria of the CMA laboratory accreditation process, and to serve as a routine quality assessment tool. PMID- 12770635 TI - Foot kinematics and kinetics during adolescent gait. AB - Gait analysis models typically analyze the ankle joint complex and treat the foot as a rigid segment. Such models are inadequate for clinical decision making for patients with foot impairments. While previous multisegment foot models have been presented, no comprehensive kinematic and kinetic databases for normal gait exist. This study provides normative foot joint angles, moments and powers during adolescent gait. Eighteen subjects were evaluated using 19 retroreflective markers, six cameras, a pressure platform and a force plate. A nine-segment model determined 3D angles, 3D moments, and powers in eight joints or joint complexes. A complete sets of sagittal, coronal and frontal plane results are presented. Results indicate that single link models of the foot significantly overestimate ankle joint powers during gait. Understanding normal joint kinematics and kinetics during gait will provide a baseline for documenting impairments in patients with foot disorders. PMID- 12770637 TI - Temporospatial and kinematic gait alterations during treadmill walking with body weight suspension. AB - Our purpose was to analyze the effects of selected levels of body weight support (BWS) on lower extremity kinematics of normal subjects at a predetermined treadmill speed. Seventeen non-disabled volunteers walked on a treadmill at 1.25 ms(-1). Temporospatial and kinematic data were collected while various support levels were applied (Minimal, 10, 30, 50 and 70% BWS). Compared to 10% BWS, significant temporospatial and kinematic changes were induced by 50 and 70% BWS. Fewer differences were induced by 30% BWS compared to 10% BWS. We concluded that gait patterns of unimpaired subjects are significantly changed by 50 and 70% BWS. PMID- 12770636 TI - Fluoroscopic and gait analysis of the functional performance in stair ascent of two total knee replacement designs. AB - Understanding total knee replacement mechanics and their influence on patient mobility requires accurate analysis of knee joint kinematics and traditional full body kinematics and kinetics. Three-dimensional fluoroscopic and gait analysis techniques were carried out on patients with either mobile bearing or posterior stabilized knee prostheses during stair ascent. Statistically significant correlation was found between knee flexion at foot strike and the position of the mid-condylar contact points, and between maximum knee adduction moment and corresponding lateral trunk tilt. A more complete and powerful assessment of the functional performances of different TKR designs can be performed in-vivo by combining gait and fluoroscopic analyses. PMID- 12770638 TI - Movement strategies for head stabilization during incline walking. AB - Changes in body orientation with respect to space during incline walking can alter vestibular information requiring a different solution to the problem of head stabilization. Eleven young adults walked along a level walkway, and ascended and descended an inclined surface. Head, neck and trunk angular positions in space were collected. Changes in the gravitoinertial vector imposed by the inclined surface, produced concomitant changes in body segment orientation that decreased head stability during the inclined walking tasks. Head, neck and trunk segments were least stable while ascending the incline creating the greatest challenge to head stability during this task. Movement strategies reflected adjustments of head-neck and neck-trunk patterns to accommodate changes in the gravitoinertial vector and insure balance of the head over the trunk. PMID- 12770639 TI - Anticipatory activation of postural muscles associated with bilateral arm flexion in subjects with different quiet standing positions. AB - We investigated changes in activation timing and magnitude of the postural muscles according to initial standing positions. The subjects were divided into three groups depending on the position of the center of foot pressure (CFP) during quiet standing, namely backward, middle, and forward. Subjects maintained standing postures at various CFP positions in the anteroposterior direction, and then started bilateral arm movement at their own pace. The activation magnitude of the biceps femoris (BF) and erector spinae (ES) did not differ among any of the initial CFP positions. In only the BF, the preceding action to the anterior deltoid (AD) was clearly observed at more forward CFP positions in the order of the forward, middle and backward groups. Between initial CFP positions adjacent to quiet standing posture, the smallest change was observed in the preceding activation time of the BF. Significant correlation was observed between the background activity and activation time in both the BF and ES. PMID- 12770640 TI - The complementary role of the plantarflexors, hamstrings and gluteus maximus in the control of stance limb stability during gait. AB - This paper focuses on the contributions of the gluteus maximus, biceps femoris, gastrocnemius and soleus in maintaining the stability of the stance limb in the sagittal plane during the mid-stance phase of gait. In the absence of any one of these muscles, the potential compensatory changes in muscle activation are explored, the aim being to restore stability to the stance limb. The investigation was carried out by integrating musculoskeletal modelling, forward simulation and optimization techniques. We concluded that maintenance of stance limb stability requires a subtle interplay of muscle activations. Weakness in a single muscle is unlikely to be adequately compensated for by increasing or decreasing the activation of one muscle alone. PMID- 12770641 TI - The effects of improved strength on obstacle negotiation in community-living older adults. AB - Poor mobility has been associated with age-related deterioration in muscle strength. While previous work has examined the effects of improved strength on level walking, we have quantified the effects of a resistance-training program on obstructed gait tasks using biomechanical-dependent measures. Forty-five community-dwelling participants aged 62 years or older were randomised to either a control (n=16) or experimental group (n=29). The experimental subjects exercised for 24 weeks on a progressive resistance-training program designed to improve lower body strength. Dynamic strength was assessed at weeks 0 and 24 as well as specific laboratory gait kinetics and kinematics during stepping over an obstacle and negotiation of a raised surface set at 10, 20 and 30% of each subject's leg length. Significant strength improvements (P<0.05), ranging between 197 and 285%, were recorded in the experimental group. The strength gains in the experimental group were accompanied by significant increases in obstacle-crossing stride velocity (range 5.5-15.5%) due to increases in stride length and decreases in stride duration for both gait tasks. Significant changes in the peak vertical and anterior-posterior ground reaction forces as well as in kinematic variables associated with a safe obstacle traverse such as vertical obstacle heel clearance, limb flexion at obstacle crossing, horizontal foot placement and vertical landing velocity resulted in an improved crossing strategy in the experimental subjects. These findings provide evidence of significant improvements in obstructed gait function of community-living older adults associated with a systematic resistance-training program. PMID- 12770642 TI - The degree of unsaturation of dietary fatty acids and the development of atherosclerosis (review). AB - Atherosclerosis is the principal contributor to the pathogenesis of myocardial and cerebral infarction, gangrene and loss of function in the extremities. It results from an excessive inflammatory-fibroproliferative response to various forms of insult to the endothelium and smooth muscle of the artery wall. Atherosclerotic lesions develop fundamentally in three stages: dysfunction of the vascular endothelium, fatty streak formation and fibrous cap formation. Each stage is regulated by the action of vasoactive molecules, growth factors and cytokines. This multifactorial etiology can be modulated through the diet. The degree of unsaturation of dietary fatty acids affects lipoprotein composition as well as the expression of adhesion molecules and other pro-inflammatory factors, and the thrombogenicity associated with atherosclerosis development. Thus, the preventive effects of a monounsaturated-fatty acid-rich diet on atherosclerosis may be explained by the enhancement of high-density lipoprotein-cholesterol levels and the impairment of low-density lipoprotein-cholesterol levels, the low density lipoprotein susceptibility to oxidation, cellular oxidative stress, thrombogenicity and atheroma plaque formation. On the other hand, the increase of high-density lipoprotein cholesterol levels and the reduction of thrombogenicity, atheroma plaque formation and vascular smooth muscle cell proliferation may account for the beneficial effects of polyunsaturated fatty acid on the prevention of atherosclerosis. Thus, the advantages of the Mediterranean diet rich in olive oil and fish on atherosclerosis may be due to the modulation of the cellular oxidative stress/antioxidant status, the modification of lipoproteins and the down-regulation of inflammatory mediators. PMID- 12770643 TI - Detection and quantification of biotinylated proteins using the Storm 840 Optical Scanner. AB - The use of the avidin-biotin interaction is becoming an increasingly common method for the detection of proteins. The use of fluorescence detection with avidin-biotin systems has the potential to greatly increase both the sensitivity and linearity of this type of analysis. In this report, three fluorescent systems were tested for their ability to detect biotinylated polypeptides in purified and complex biological samples. These systems include a Neutravidin-Alexa Fluor430 conjugate, an avidin-horseradish peroxidase conjugate with the ECL-Plus detection system, and an avidin-alkaline phosphatase conjugate with the ECF detection system. Biotinylated molecular weight standards, biotinylated bovine serum albumin, and rat liver homogenate were resolved by SDS-PAGE gel electrophoresis and transferred to polyvinyldifluoride membrane. Biotinylated polypeptides were then visualized on the Storm840 optical scanner. The Neutravidin-Alexa Fluor430 conjugate exhibited the lowest sensitivity, but displayed high linearity. The avidin-horseradish peroxidase and avidin-alkaline phosphatase conjugates, when combined with appropriate fluorescent substrates, exhibited much higher fluorescence, with the avidin-alkaline phosphatase ECF system displaying the highest sensitivity. All systems demonstrated an ability to reliably detect and quantify biotinylated polypeptides in purified as well as complex samples, given careful attention to conditions optimized for each system. PMID- 12770644 TI - Dietary fibres ameliorate decreased synthesis of heparan sulphate in streptozotocin induced diabetic rats. AB - The role of dietary fibers in diabetes has been studied by several workers. Long term dietary treatment with increased amounts of fiber-rich low-glycaemic index natural foods improves blood glucose and reduces the number of hypoglycemic events in type I diabetic patients. On the other hand Rohrbach and Martin and Cohen and Surma described changes in the general and biochemical structure of renal tissues such as the glomerular basement membranes. One of these changes was the reduction and undersulfation of the glycoconjugate and glycosaminoglycan heparan sulfate, which plays an important role in renal structure and function. The purpose of the present study was to determine specific effects of two types of dietary fiber on the composition of kidney glycoconjugates in an animal model of diabetes type I. Streptozotocin-treated diabetic rats were fed either a control diet or diets containing 10% wheat bran (insoluble dietary fiber) or 5% guar gum (soluble dietary fiber). Effects of these fibers on glycaemic control and nephropathy were assessed using previously described methodologies. The effect of dietary fiber in the glycoconjugate composition of kidneys of control and diabetic animals was studied by estimating their total hexose content, sulfated glycosaminoglycans, hexosamines and uronic acids. The activities of enzymes that participate in the synthesis of saccharides and glycoconjugates (L glutamine-fructose-6-phosphate aminotransferase) and their degradation (N-acetyl beta-glucosaminidase and beta-glucuronidase) were also evaluated. Results indicated that both soluble and insoluble dietary fibers ameliorated a significant increase in the activity of GFAT. Heparan sulfate was also isolated and quantified. Results indicated that the renal content of heparan sulfate decreased in diabetic animals and that this decrement was ameliorated by the ingestion of both soluble and insoluble fiber in the diet. PMID- 12770645 TI - Vitamin E supplementation in the mitigation of carbon tetrachloride induced oxidative stress in rats. AB - Oxidative stress is implicated in the pathophysiology of a number of chronic diseases including atherosclerosis, diabetes, cataracts and accelerated aging. The aim of this study was to elucidate the protective role of vitamin E supplementation when oxidative stress is induced by CCl4 administration, using the rat as a model. Rats were fed diets for four weeks either with or without dl alpha-tocopherol acetate supplementation. Half of the rats (n = 9) from each of the diet groups were then challenged with CCl4 at the completion of the four week diet period. Plasma levels of 8-iso-PGF(2alpha), antioxidant micronutrients and antioxidant enzyme activities were measured to examine changes in oxidative stress subsequent to the supplementation of dl-alpha-tocopherol in the diet. Plasma alpha-tocopherol (vitamin E) concentrations were higher for the groups supplemented with dl-alpha-tocopherol acetate, however the supplemented diet group that was subsequently challenged with CCl4 had significantly lower (p <0.001) plasma alpha-tocopherol concentration than the dl-alpha-tocopherol acetate diet group that was not challenged with CCl4. Total plasma 8-iso PGF(2alpha) concentration was elevated in diet groups challenged with CCl4, however, the concentration was significantly lower (p <0.001) when the diet was supplemented with dl-alpha-tocopherol acetate. The antioxidant enzymes were not influenced by either dietary alpha-tocopherol manipulation or by the inducement of oxidative stress with CCl4. Plasma concentrations of trans-retinol (vitamin A) were reduced by CCl4 administration in both the dl-alpha-tocopherol acetate supplemented and unsupplemented diet groups. The results of this study indicate that dl-alpha-tocopherol acetate supplementation was protective of lipid peroxidation when oxidative stress is induced by a pro-oxidant challenge such as CCl4. PMID- 12770647 TI - Effect of selenium-enriched broccoli diet on differential gene expression in min mouse liver(1,2). AB - Multiple intestinal neoplasia (Min) mice are a good model for investigating the effects of dietary alterations in a genetic model for intestinal cancer. Previous studies have shown that selenium-enriched broccoli effectively reduces colon cancer susceptibility. Although colon cancer cells mainly metastasize to the liver, little is known about the effects of selenium-enriched broccoli on gene expression in mouse liver. To better understand the protective role for selenium enriched broccoli in tumorigenesis, a gene profile of the mouse liver was analyzed. Mice were fed either 0.11 mg selenium/kg control diet or 2.1 mg selenium/kg selenobroccoli diets for 10 weeks. Use of mouse pathway finder-1 GEArrays revealed that selenium-enriched broccoli moderately increased ikBalphakappaB, hsp86, gadd45 gene transcripts. In addition, analysis of the binding of liver nuclear proteins to (32)P-labeled probes demonstrated that selenium-enriched broccoli enhanced the binding of transcription factor p53, NFkappaB, AP-1 to their cis-acting elements. Collectively, these results suggest for the first time that selenium-enriched broccoli activates certain pro apoptotic genes linked to p53, NFkappaB and stress signal pathways in response to "danger signals" such as tumorigenesis. PMID- 12770646 TI - Effects of mitochondrial uncoupling on adipocyte intracellular Ca(2+) and lipid metabolism. AB - Previous data from this laboratory demonstrate that increased intracellular Ca(2+) ([Ca(2+)]i) coordinately regulates human and murine adipocyte lipid metabolism by stimulating lipogenesis and inhibiting lipolysis. However, recent data demonstrate metabolic uncoupling increases [Ca(2+)]i but inhibits lipogenesis by suppressing fatty acid synthase (FAS) activity. Accordingly, we have evaluated the interaction between mitochondrial uncoupling, adipocyte [Ca(2+)]i, and adipocyte lipid metabolism. Pretreatment of 3T3-L1 cells with mitochondrial uncouplers (DNP or FCCP) amplified the [Ca(2+)]i response to depolarization with KCl by 2-4 fold (p <0.001), while this increase was prevented by [Ca(2+)]i channel antagonism with lanthanum. Mitochondrial uncouplers caused rapid (within 4hr) dose-dependent inhibition of FAS activity (p <0.001), while lanthanum caused a further additive inhibition. The suppression of FAS activity induced by uncoupling was reversed by addition of ATP. Mitochondrial uncouplers increased FAS expression significantly while [Ca(2+)]i antagonism with lanthanum decreased FAS expression (P <0.001). In contrast, mitochondrial uncouplers independently inhibited basal and isoproterenol-stimulated lipolysis (20-40%, p <0.001), while this inhibition was fully reversed by lanthanum. Thus, mitochondrial uncoupling exerted short-term regulatory effects on adipocyte [Ca(2+)]i and lipogenic and lipolytic systems, serving to suppress lipolysis via a Ca(2+) -dependent mechanism and FAS activity via a Ca(2+)-independent mechanism. PMID- 12770648 TI - Patterns of food intake and self-selection of macronutrients in rats during short term deprivation of dietary zinc. AB - Although it has been known for more than 50 years that zinc (Zn) deficiency regularly and consistently causes anorexia in many animal species, the basic mechanism(s) that cause this phenomenon still remain(s) an enigma. The following studies describe feeding behavior in the early stages of zinc deficiency in the rat model. In one experiment, we used computerized feeding monitors that measured the intake of individual rats at 10-min intervals over 24-hr periods. Male rats were acclimated to the cages and fed a Zn-adequate egg-white-based diet, or a similar diet with <1.0 mg Zn/kg. Food intake was monitored for seven, consecutive 24-hr periods. The 24-hr food intake pattern of the Zn-deprived rats did not differ from the controls; they simply ate less food, mainly during the night hours, with no differences between groups during the day. Although Zn-deprived rats ate less food than controls, the percentage of total diet consumed during night and day did not differ between groups. In another experiment, we simultaneously offered male rats three isocaloric diets with different macronutrient compositions and with or without adequate Zn, and measured the amount of each diet selected during seven, 24-hr periods. The three diets contained either 57% protein from egg white, 30% fat from soybean oil, or 80% carbohydrate from a combination of starch, hydrolyzed starch, and sucrose. For the first four days on experiment, rats selected similar amounts of each diet. Then the Zn-deprived rats began to select only 50% as much of the protein diet as the controls. Similar results were obtained when the data were expressed on the basis of each macronutrient as a percentage of the total diet selected. Zn deprived rats selected a diet that contained 8% protein, 73% carbohydrate, and 6% fat while the Zn-adequate rats selected 12% protein, 69% carbohydrate, and 6% fat. Fat intake was not affected by Zn-deprivation. The results confirm our previous findings, and are discussed in terms of Zn-deprivation blunting the pathways of signal transduction that involve the peptide hormones known to affect food intake regulation. PMID- 12770650 TI - Of plague and "pestilence". PMID- 12770651 TI - Interleukin balance and early recovery from anesthesia in elderly surgical patients exposed to beta-adrenergic antagonism. AB - STUDY OBJECTIVE: To determine whether proinflammatory and antiinflammatory cytokines, as measured in blood specimens, would correlate with improved SF-36 physical composite scores observed in elderly surgical patients who were administered perioperative atenolol. DESIGN: Post hoc analysis of data from a randomized clinical study. SETTING: Department of Anesthesiology, Mount Sinai Medical School, New York. PATIENTS: 59 ASA physical status II, III, and IV patients > or =65 years of age, who were scheduled for major elective noncardiac surgery. INTERVENTIONS: Patients were randomized to one of three anesthetic regimens to receive 1) perioperative management without beta-adrenergic antagonism, 2) preoperative and postoperative administration of atenolol, or 3) intraoperative atenolol as a major component of the anesthetic regimen. MEASUREMENTS AND MAIN RESULTS: Blood samples were drawn perioperatively at seven different time points. Interleukin-1 beta, interleukin-6, interleukin-1ra, and interleukin-10 were measured using enzyme-linked immunosorbent assay (ELISA) kits. Also, recovery from anesthesia and physical/mental well-being (SF-36 questionnaire) were determined perioperatively. Compared with control patients, atenolol-treated patients experienced improved postoperative physical well-being, which paralleled the previously reported faster recovery from anesthesia and a decreased need for perioperative analgesics. Improved postoperative physical well being of atenolol-treated patients was specifically caused by an ameliorated bodily pain score, a major component of the physical composite score of the SF-36 questionnaire. The cytokine response of these elderly surgical patients was similar to that of younger patients, and the perioperative profile of proinflammatory and antiinflammatory cytokines was not affected by atenolol. CONCLUSIONS: Perioperative administration of atenolol to elderly surgical patients markedly improves physical sense of well-being, which coincides with improved postoperative pain control and decreased analgesic requirements. This improvement experienced by patients receiving atenolol is not related to alterations in perioperative cytokine response. PMID- 12770652 TI - The dilemma of immediate preoperative hypertension: to treat and operate, or to postpone surgery? AB - STUDY OBJECTIVE: To evaluate the efficacy and complications of immediate preoperative reduction of arterial blood pressure (BP) in patients with well controlled hypertension but with diastolic blood pressure (DBP) between 110 and 130 mmHg on arrival at the operating room (OR). DESIGN: Prospective, randomized, large-sample study. SETTING: University-affiliated, 550-bed community hospital. PATIENTS: 989 patients with well-controlled hypertension, who were scheduled for surgery, and who had no previous myocardial infarction, unstable or severe angina pectoris, renal failure, pregnancy induced hypertension, left ventricular hypertrophy, previous coronary revascularization, aortic stenosis, preoperative dysrhythmias, conduction defects, or stroke. INTERVENTIONS: Patients with DBP between 110 and 130 mmHg were randomly allocated to two groups: 400 patients in the control group and 589 patients serving as the study group. The control group had their surgery postponed and they remained in hospital for BP control, and the study patients received 10 mg of nifedipine intranasally delivered. The patients were observed for cardiovascular and neurological complications during the intraoperative period and over the first three postoperative days. MEASUREMENTS AND MAIN RESULTS: The two groups were similar in age, gender, type of surgery, duration of anesthesia, and intraoperative fluid administration. There were no statistically significant differences in postoperative complications. The hospitalization time was considerable shorter in the study group than in the control group. CONCLUSIONS: Immediate preoperative reduction of DBP with intranasal nifedipine is safe in patients with well-controlled arterial hypertension but they presented with severe to very severe hypertension for patients in the OR. We were able to avoid unnecessary surgery postponement and attendant costs. PMID- 12770653 TI - Recombinant factor VIIa for control of hemorrhage: early experience in critically ill trauma patients. AB - STUDY OBJECTIVE: To examine our institutional experience with recombinant Factor VIIa (rFVIIa) as a treatment for exsanguinating hemorrhage in critically ill trauma patients. DESIGN: Retrospective case review. SETTING: A specialized trauma and critical care hospital, serving as the quaternary referral center for trauma and surgical shock in the state of Maryland. PATIENTS: All patients with diffuse coagulopathy and impending exsanguination, given rFVIIa in an effort to control life-threatening hemorrhage. Patients were in the intensive care unit (ICU) or operating room (OR) and included both acute admissions and late-stage patients with multiple organ system failure. INTERVENTIONS: Patients of interest were those that had received rFVIIa. MEASUREMENTS: Examination of medical records, including pharmacy data, laboratory results, and the institutional trauma registry. MAIN RESULTS: Administration of rFVIIa contributed to successful control of hemorrhage in three of five patients. Failure in two patients was mostly likely due to overwhelming shock and acidosis. CONCLUSIONS: Administration of rFVIIa shows promise in the treatment of exsanguinating hemorrhage. Prospective, controlled clinical trials of this therapy are strongly recommended. PMID- 12770654 TI - Body Weight-Related ionized hypomagnesemia in pediatric patients undergoing cardiopulmonary bypass for surgical repair of congenital cardiac defects. AB - STUDY OBJECTIVES: To examine the serial time course of perioperative plasma ionized magnesium (iMg(2+)) concentrations and to analyze the plasma iMg(2+) concentrations in children with different body mass who were undergoing open heart surgery. DESIGN: Randomized, single-blinded study. SETTING: University affiliated hospital of an academic medical institution. PATIENTS: 38 children undergoing open-heart surgery. INTERVENTIONS: Patients were divided into three groups according to their body mass: Group 1 (n = 12) <10 kg, Group 2 (n = 13) 10 kg to 20 kg, and Group 3 (n = 13) >20 kg. MEASUREMENTS: The relationship of iMg(2+) among the three groups of different body mass were analyzed at five different time intervals during the operation: induction of anesthesia, 5 minutes and 30 minutes after the onset of cardiopulmonary bypass (CPB), the beginning of rewarming, and the end of surgery. MAIN RESULTS: iMg(2+) levels at 5 minutes after onset of CPB in patients weighing less than 20 kg (Groups 1 and 2) differed with those weighing more than 20 kg (Group 3) (p = 0.007 and 0.013). However, there was no difference in the iMg(2+) levels between Groups 1 and 2 (p = 0.993). In addition, iMg(2+) levels at 5 minutes after onset of bypass correlated well (r(2) = 0.66) in children with body mass less than 20 kg. CONCLUSIONS: Low levels of ionized magnesium is an important finding in patients at the onset of CPB, which correlates well with the body mass of patients weighing less than 20 kg, and could be predicted by the regression curve. Based on these findings, hypomagnesemia can be prevented during CPB. PMID- 12770655 TI - A study of anesthetic drug utilization in different age groups. AB - STUDY OBJECTIVE: To determine anesthetic drug utilization in different age groups. DESIGN: Retrospective, automated, intraoperative database study. SETTING: Tertiary care medical center. MEASUREMENTS: 30,842 noncardiac general anesthesia case records between January 1991 and July 1997 were studied. We investigated the effect of age on anesthetic requirements for fentanyl (F), midazolam (M), thiopental sodium (T), propofol (P), isoflurane (I), and nitrous oxide (N). Because drugs are not given in isolation we looked at the most common drug combinations, IFNTM, IFNPM, INFT, and PFNM. Regression analyses on log transformed drug dosages were used to test the significance of age on individual requirements. RESULTS: In each of the above anesthetic drug combinations, reduced doses of fentanyl, propofol, midazolam, thiopental, and isoflurane were used with increasing age. Fentanyl, propofol, thiopental, and isoflurane showed a 10%, 8%, 6%, and 4% reduction in dose per decade of age, respectively, from age of maximum dose to age 80 years. CONCLUSIONS: In clinical practice, increasing age results in decreased anesthetic drug administration. The mechanism of this observation needs to be determined. PMID- 12770657 TI - Physicians' perceptions of minimum time that should be saved to move a surgical case from one operating room to another: internet-based survey of the membership of the Association of Anesthesia Clinical Directors (AACD). AB - STUDY OBJECTIVE: Moving the last case of the day from one operating room (OR) to another OR can increase OR efficiency. However, there is a penalty cost for moving a case. The goal of the study was to measure perceptions of the minimum time that needs to be saved for it to be worthwhile to move a case from a late running OR to another OR. DESIGN: Internet-based survey of the Association of Anesthesia Clinical Directors (AACD) and/or attendees at one of its courses. As subjects completed the computer-assisted survey, answers to test questions were checked immediately to ensure respondents understood the relevant concepts. MEASUREMENTS: Respondents were asked to complete the statement: "I would move the case if I would expect to save ____ hours of overutilized OR time." MAIN RESULTS: 234 E-mail invitations to complete the survey were transmitted. Of that number, 87 completed surveys were returned. Respondents were physicians, mostly from the United States. The 25th, 50th, and 75th percentiles of the penalty cost were 1.0 hour of overutilized OR time. The 95% confidence intervals were 0.5 to 1.0 hour for the 25th percentile, 1.0 to 1.0 hour for the 50th percentile, and 1.0 to 2.0 hours for the 75th percentile. There was no significant correlation between the penalty cost and the number of ORs at the respondent's facility, number of times the survey was submitted until it was completed correctly, or total number of errors in responses. CONCLUSIONS: Members of the AACD perceive the penalty cost for moving a case to be 1 hour. PMID- 12770656 TI - Local anesthesia and midazolam versus spinal anesthesia in ambulatory pilonidal surgery. AB - STUDY OBJECTIVE: To evaluate two anesthetic techniques, namely, local anesthesia with sedation, and spinal anesthesia, with respect to recovery times, postoperative side effects, pain scores, patient satisfaction, and hospital costs for ambulatory pilonidal disease surgery. DESIGN: Prospective, randomized study. SETTING: University Hospital of Pamukkale. PATIENTS: 60 consenting patients scheduled for pilonidal disease operation with Limberg flap technique. INTERVENTION: Patients were randomly allocated into two groups: Group 1 (n = 30) received spinal anesthesia with hyperbaric bupivacaine 1.5 mL 0.5%, and Group 2 (n = 30) received local infiltration with a 50-mL mixture containing 10 mL bupivacaine 0.5%, 10 mL prilocaine HCl 2%, and 30 mL isotonic solution with 1:200,000 epinephrine in combination with intravenous (i.v.) midazolam sedation. MEASUREMENTS: Perioperative and postoperative side effects, patient satisfaction, preoperative visual analog scale (VAS) pain scores, and VAS scores from the fourth hour postoperatively until the seventh day were assessed. Anesthesia, operation, surgery, and total hospital time, and costs (drug, resources, and labor) were recorded. MAIN RESULTS: No difference was found between groups in the frequency of side effects. Urinary retention was diagnosed in two patients in the spinal anesthesia group. There was no statistical significant difference seen in satisfaction scores between groups. No statistical significance in VAS pain scores between groups was noted except for the fourth postoperative hour values. The average time spent in the operating room (OR) was greater in the spinal anesthesia group. All Group 2 patients achieved fast-tracking criteria in the OR and were able to bypass the postanesthesia care unit (PACU). Total hospital time and total cost were significantly higher in the spinal anesthesia group than local anesthesia-sedation group (p < 0.05). CONCLUSION: The use of local anesthesia-sedation for ambulatory anorectal surgery resulted in a shorter hospital time, lower hospital costs, and no side effects compared with spinal anesthesia. PMID- 12770658 TI - Anesthetic management in fibrodysplasia ossificans progressiva (FOP): a case report. AB - Fibrodysplasia ossificans progressiva (FOP) is a rare congenital disorder of connective tissue, which has significant implications on anesthetic management of affected patients, e.g., application of difficult airway algorithm, avoiding any trauma during venous cannulation. FOP has received very limited reviews in the published anesthesia literature. We describe perioperative care of a 21-year-old female with established diagnosis of FOP presenting for hysteroscopy followed by dilatation and curettage. Multiorgan abnormalities of FOP are reviewed and available anesthetic options are analyzed and compared. PMID- 12770659 TI - Airway management of a patient with Weill-Marchesani syndrome. AB - Weill-Marchesani syndrome is a genetically determined rare systemic connective tissue disorder consisting of brachymorphy, brachydactyly, aortic stenosis, and ophthalmologic abnormalities, such as ectopia lentis, microspherophakia, and secondary glaucoma. The primary lesion is fibrous tissue hyperplasia. Airway control and intubation may be difficult in patients with WMS because of stiff joints, poorly aligned teeth, and maxillary hypoplasia with a narrow palate. We describe the successful airway management of a patient with WMS and laryngeal stenosis, using an intubating laryngeal mask following failed conventional laryngoscopic intubation. PMID- 12770660 TI - Horner's syndrome following very low concentration bupivacaine infusion for labor epidural analgesia. AB - Horner's syndrome is a complication of epidural analgesia and anesthesia, encountered more commonly in pregnant women than in other patients. Previous reports described the appearance of Horner's syndrome following epidural injection of concentrated local anesthetic solutions. We report unilateral Horner's syndrome occurring in the setting of lumbar epidural analgesia for labor with a very low local anesthetic concentration (bupivacaine 0.04%) in an epidural infusion. We discuss the possible factors that could have contributed to this occurrence despite the extremely dilute concentration of local anesthetic used for analgesia. PMID- 12770661 TI - Use of a human polymerized hemoglobin solution as an adjunct to acute normovolemic hemodilution during complex abdominal aortic reconstruction. AB - Oxygen-carrying hemoglobin (Hb) solutions are under intense investigation as an alternative to allogeneic red cell transfusion during surgery, with or without acute normovolemic hemodilution. We present a case in which an investigational Hb solution was used as an adjunct to acute normovolemic hemodilution, and as a replacement for surgical blood loss in a patient undergoing complex aortic reconstruction with a large blood loss. PMID- 12770662 TI - Human immunodeficiency virus in the parturient. AB - Over the last 20 years, the acquired immune deficiency syndrome (AIDS) has grown from a small case series of Pneumocystis carinii infection in four homosexual men to one of the major health problems facing the world today. In the next 5 years, human immunodeficiency virus (HIV) infection is expected to kill more than 2.2 million people. In the United States, women of childbearing age constitute a large percentage of new cases of AIDS. Because of the increased prevalence of HIV in pregnant women, many anesthesiologists encounter these patients in their practice. The safety of regional neuraxial spread has been a concern in the past, nevertheless, recent analysis of the problem has shown that HIV infection in pregnancy does not contraindicate administration of regional anesthesia. PMID- 12770664 TI - Variations of coronary sinus acid-base and gas profile depending on the sampling position during bypass surgery. PMID- 12770663 TI - Neuraxial opioid-induced pruritus: a review. AB - When intrathecal and epidural opioids are administered, pruritus occurs as an unwanted and troublesome side effect. The reported incidence varies between 30% and 100%. The exact mechanisms of neuraxial opioid-induced pruritus remain unclear. Postulated mechanisms include the presence of an "itch center" in the central nervous system, medullary dorsal horn activation, and antagonism of inhibitory transmitters. The treatment of intrathecal opioid-induced pruritus remains a challenge. Many pharmacological therapies, including antihistamines, 5 HT(3)-receptor antagonists, opiate-antagonists, propofol, nonsteroid antiinflammatory drugs, and droperidol, have been studied. In this review, we will summarize pathophysiological and pharmacological advances that will improve understanding and ultimately the management of this troublesome problem. PMID- 12770665 TI - Inaccurate labeling of intravenous fluid bags: implications for anesthesiologists. PMID- 12770666 TI - Uterine rupture: signs and symptoms and the effect of labor analgesia on diagnostic accuracy. PMID- 12770667 TI - Paroxysmal dyskinesias in childhood. AB - We report on clinical features of a large series of patients with paroxysmal dyskinesias. Fourteen patients had paroxysmal kinesigenic dyskinesia, with a mean age at onset of 7.1 years. In thirteen children the condition was idiopathic and nine of them had a positive family history; the remaining one had a Chiari malformation. Response to antiepileptic drugs was good in 60% of the treated patients. Six children had paroxysmal non-kinesigenic dyskinesia, with a mean age at onset of 8.1 years. Five children were symptomatic because of cerebral palsy (two patients), basal ganglia stroke (one patient), and acute inflammatory encephalopathy (one patient); the remaining patient's condition was familial with autosomal-dominant mode of inheritance. Response to medical treatment was unsuccessful contrasting with paroxysmal kinesigenic dyskinesia. Six children had paroxysmal exercise-induced dyskinesia, with a mean age at onset of 5 years. Their condition was idiopathic, with a positive family history in four. Two of these patients had also rolandic epilepsy and writer's cramp, and the syndrome had been previously mapped to chromosome 16. Although there have been great advances in the genetics of paroxysmal dyskinesias in which an ion channel dysfunction has been hypothesized, the diagnosis is still based on clinical grounds. The precise classification of the patients with paroxysmal dyskinesias is important for therapeutic decisions. PMID- 12770668 TI - Intracranial pressure changes during intermittent CSF drainage. AB - Premature very-low-birth-weight infants with posthemorrhagic hydrocephalus are often managed with intermittent cerebrospinal fluid drainage from a ventricular reservoir. There are little data regarding intracranial pressure changes during intermittent drainage to determine the amount and frequency of cerebrospinal fluid removal or to determine the correct resistance of future programmable shunts. The objective of this study was to determine the feasibility of using a commercially available intracranial pressure transducer to measure changes in pressure associated with this procedure. Continuous intracranial pressure was measured in three infants with a transducer placed at the time of ventricular reservoir insertion. Daily reservoir taps began 48 hours after placement and intracranial pressure was monitored for 7 days. Intracranial pressure before the initial tap was comparable to levels previously reported as normal. The daily removal of 10 cc/kg of cerebrospinal fluid was sufficient to lower intracranial pressure below baseline, however it was associated with wide swings in pressure and, in one patient, sustained negative pressure. The use of direct intracranial pressure monitoring may be useful in determining the optimal amount and frequency of cerebrospinal drainage from infants with posthemorrhagic hydrocephalus managed with a ventricular reservoir, as well as determining resistance settings of subsequent programmable shunts. PMID- 12770669 TI - Predictive value of electrophysiology in children with hypoxic coma. AB - Assessment of prognosis of children in hypoxic coma is difficult. The value of clinical evaluation is often limited. The usefulness of electrophysiologic tests has been documented mostly in adults and neonates and in cases of traumatic coma. We reviewed retrospectively 39 consecutive children with nontraumatic hypoxic coma to assess the prognostic value of EEG, visual, and auditory evoked potentials. Correlation between electrophysiology and neurologic outcome after mean follow-up period of 30 months was significant (r(s) = 0.6, P < 0.001). In contrast there was no correlation between Pediatric Risk of Mortality score (PRISM) and outcome (r(s) = -0.42, P = 0.8). Combining magnetic resonance imaging with electrophysiology further enhanced their prognostic value (r(s) = 0.69, P < 0.001). Neuroimaging was highly sensitive but less specific, and electrophysiologic tests were highly specific but less sensitive. We conclude that early electrophysiology can contribute to predicting outcome in pediatric hypoxic coma. PMID- 12770670 TI - Oligohydrosis and fever in pediatric patients treated with zonisamide. AB - Zonisamide is an antiepileptic drug developed and first marketed in Japan in 1989. Cases of oligohydrosis, characterized by deficient production and secretion of sweat, were reported in children treated with zonisamide in Japan during development and in the postmarketing period. Zonisamide was approved in the United States in March 2000 for adjunctive treatment of partial seizures in adults. Searching the Food and Drug Administration's Adverse Events Reporting System, we identified six domestic cases of zonisamide-associated oligohydrosis and/or fever, all in patients 300"). Therefore, TMS and backward masking can interact in a non-linear manner, strongly interfering with early visual processing. PMID- 12770689 TI - Lack of association between a single nucleotide polymorphism within the choline acetyltransferase gene and patients with Alzheimer's disease. AB - Alterations of the cholinergic system may account for typical clinical and pathophysiological disturbances of Alzheimer's disease (AD). In particular, a marked decline of choline acetyltransferase activity (CHAT) and as a consequence of acetylcholine during the course of the disease has been described. Due to the chromosomal localization of CHAT at 10q11.23 and its possible role in the pathophysiology of AD, CHAT may represent an appropriate candidate gene conferring risk to AD. In fact, a recent study identified a functional single nucleotide polymorphism (SNP) within the first common exon of CHAT, which was associated with AD giving an odds ratio of 3.8 (Neurosci. Lett. 333 (2002) 9). Because of the potential importance of this finding we analyzed this SNP and another functional SNP within exon 9 (rs868749) of the CHAT gene using a German case control sample consisting of 242 patients with AD and 143 cognitively healthy controls. No statistically significant differences were obtained for the previously described polymorphism. In addition, the exon 9 SNP (rs868749) was not polymorphic in the studied population. We conclude that the previously identified polymorphism is not associated with AD. PMID- 12770691 TI - Cyclooxygenase-2 in the hippocampus is up-regulated in Alzheimer's disease but not in variant Alzheimer's disease with cotton wool plaques in humans. AB - We examined neuronal expression of cyclooxygenase-2, a pro-inflammatory protein, and neuron densities in the CA1-4 of the hippocampus in three cases of Alzheimer's disease with cotton wool plaques (CWP-AD), 17 cases of typical Alzheimer's disease without CWPs (tAD), and 26 normal controls. Cyclooxygenase-2 expression was significantly increased in all of the CA1-4 in tAD, but not in any subdivision in CWP-AD, compared with controls. Cyclooxygenase-2 expression in tAD was also significantly up-regulated compared with that in CWP-AD in all subdivisions. Furthermore, neuron density in the hippocampus was not significantly reduced in CWP-AD cases compared with controls despite remarkable intra- and extraneuronal Abeta deposition. These findings suggest that unknown factors besides Abeta deposition are necessary for the cyclooxygenase-2 up regulation and neurodegeneration in Alzheimer's disease. PMID- 12770692 TI - Pharmacological characterization of recombinant bovine alpha3beta4 neuronal nicotinic receptors stably expressed in HEK 293 cells. AB - In these studies, [(3)H]epibatidine is used as the radioligand to characterize recombinant bovine alpha3beta4 nicotinic acetylcholine receptors (nAChRs) expressed in HEK 293 cells. Specific binding reaches equilibrium quickly and is saturable with a K(d) value of 0.66 nM. The affinities of the several cholinergic agents were determined, including nicotine (K(i), 0.5 microM), cytisine (K(i), 0.5 microM), carbachol (K(i), 4.1 microM), dihydro-(beta)-erythroidine (K(i), 43.5 microM), d-tubocurarine (K(i), 0.1 microM), 1,1-dimethyl-4 phenylpiperazinium (K(i), 0.5 microM), decamethonium (K(i), 175 microM) and methyllycaconitine (K(i), 0.4 microM). These studies show that the pharmacological characteristics of recombinant bovine alpha3beta4 nAChRs are similar to native bovine alpha3beta4* nAChRs, and indicate that the alpha5 subunit, if present in the native nAChRs, does not affect ligand affinity. PMID- 12770693 TI - ATP-sensitive potassium channels in rat primary afferent neurons: the effect of neuropathic injury and gabapentin. AB - ATP-sensitive potassium (K(ATP)) currents were examined in dorsal root ganglion neurons from neuropathic and control rats using whole-cell voltage clamp recordings. K(ATP) channel openers (diazoxide and pinacidil) enhanced, and the blocker glibenclamide inhibited an outward current in control neurons in a manner dependent on the pipette ATP concentration. Analysis of reversal potentials showed that this current is carried by K(+) ions. Outward current in cells from rats with peripheral nerve injury was not sensitive to modulators of K(ATP) channels. Gabapentin, a putative K(ATP) channel opener, had minimal effect on currents in either group of neurons. We conclude that normal primary afferent neurons express K(ATP) channels that conduct current which is eliminated by peripheral nerve injury. Gabapentin does not affect this current significantly. PMID- 12770694 TI - Involvement of N-methyl-D-aspartate receptors in post-hypoxic depression of the dorsomedial medulla in cats. AB - Pressor and sympathoexcitatory responses induced by microinjection of glutamate (Glu) into the dorsomedial medulla (DM) were depressed after hypoxia in anesthetized cats. This study was undertaken to investigate which Glu receptor subtypes would be involved in the post-hypoxic depression. Hypoxia was induced by inhalation of a 5% O(2) and 95% N(2) gas mixture. The pressor and sympathoexcitatory responses to microinjections of N-methyl-D-aspartic acid (NMDA) or alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) into the DM were significantly depressed after hypoxia, suggesting the involvement of both NMDA and AMPA receptors. However, pretreatment with kynurenic acid or DL-2 amino-5-phosphonopentanoic acid, but not 6-cyano-7-nitroquinoxaline-2,3-dione, 5 min before hypoxia could effectively prevent the post-hypoxic depression of Glu induced responses. These results further suggest that post-hypoxic depression of Glu-induced responses in the DM was predominately mediated by NMDA receptors. PMID- 12770695 TI - Effects of working memory demands on frontal slow waves in time-interval reproduction tasks in humans. AB - Very few studies have examined the event-related potentials associated with the retention of temporal information for later use. In the present experiment, event related potentials were recorded during two duration reproduction tasks in which a delay was introduced between the encoding and the reproduction phases. Furthermore, working memory demands were varied during the delay (number of durations to be memorized and degree of manipulation of the durations). These variables had no significant effect on the amplitude of the contingent negative variation observed during duration encoding. By contrast, the amplitude of the slow wave recorded over middle frontal regions during the delay increased with working memory demands, which confirms the key role played by these frontal regions in the cognitive processes engaged during the retention of duration in working memory. PMID- 12770696 TI - Cold and mechanical allodynia in both hindpaws and tail following thoracic spinal cord hemisection in rats: time courses and their correlates. AB - We assessed (1) the time courses of cold and mechanical allodynia in both hindpaws and the tail, and (2) the relationship of the allodynia signs between different sites following spinal cord hemisection. Under enflurane anesthesia, rats were subjected to spinal hemisection at T13. The hemisected rats exhibited a significant increase in mechanical and cold allodynia signs of both hindpaws and the tail for 22-26 weeks postoperatively. In addition, mechanical allodynia signs were significantly correlated not only between the ipsilateral and the contralateral hindpaws, but also between the hindpaws and the tail. These results suggested that cold and mechanical allodynia developed extensively and lasted for a long time following spinal cord hemisection, and mechanical allodynia shown at different sites may be induced at least in part by common generating mechanisms. PMID- 12770697 TI - Distinct effects of amantadine and memantine on dopaminergic transmission in the rat striatum. AB - Striatal glutamatergic inputs are known to participate in the modulation of dopaminergic transmission. Accordingly, the non-competitive N-methyl-D-aspartate receptor antagonists memantine and amantadine increase striatal dopamine levels, the latter being widely used in Parkinson's disease therapy. Based on our previous work revealing increased function of dopamine receptors and dopamine transporter after amantadine treatment, we studied the effects of repeated memantine administration on dopaminergic neurotransmission. On rat striatal membranes, dopamine-stimulated [(35)S]GTPgammaS binding was significantly reduced (20%) after 2 days injection with memantine (20 mg/kg per day, i.p.) but not after longer treatments (4 or 7 days). Evaluation of [(3)H]SCH 23390 and [(3)H]spiperone specific bindings only revealed a significant increase in D1 receptor density after 4 or 7 days treatment. Finally, none of these treatments were found to change the activity of the neuronal dopamine transporter in striatal synaptosomes. This shows that amantadine and memantine differentially affect striatal dopaminergic transmission, which could indicate that these two related aminoadamantanes display distinct pharmacodynamic properties. PMID- 12770698 TI - Absence of association between Alzheimer disease and the regulatory region polymorphism of the PS2 gene in an Italian population. AB - Alzheimer disease (AD) is the most common neurodegenerative disorder of aging. Identifying novel AD genetic risk factors is important for understanding its pathogenesis. A recent study demonstrated that the deletion of adenosine in the promoter region of the presenilin 2 gene (PS2) is a susceptibility factor for early-onset AD. The objective of our study was to test the possibility that this variation is associated with AD in the Italian population. A case-control association study was performed, using 200 sporadic AD cases and 160 normal controls matched by age, gender and ethnicity. The current study does not support the notion that the polymorphism in the PS2 gene constitutes a risk factor for either late-onset or early-onset AD, which means that other genetic factors play a role in the development of AD in the Italian population. PMID- 12770699 TI - Serum cholesterol levels modulate long-term efficacy of cholinesterase inhibitors in Alzheimer disease. AB - The clinical, genetic or biological variables which regulate long-term efficacy of cholinesterase inhibitors (ChEIs) in Alzheimer disease (AD) are still unknown and it is not possible to predict who will benefit from the treatment. In this study we showed that high cholesterol levels correlated with faster decline at 1 year follow-up in AD patients on ChEIs. These findings suggest that serum cholesterol is a modulating factor of treatment response and additional therapies aimed at reducing treatable high cholesterol levels may represent an alternative strategy to improve ChEIs efficacy and slow down disease progression over time. PMID- 12770700 TI - Overeating, alcohol and sucrose consumption decrease in CB1 receptor deleted mice. AB - Administration of the cannabinoid CB1 receptor antagonist SR141716 (3-10 mg/kg i.p.) abolished neuropeptide Y-induced overeating and significantly reduced ethanol and sucrose intake in CB1 wild-type (+/+) mice. In CB1 receptor knockout (-/-) mice, neuropeptide Y totally lost its capacity to increase food consumption. Similarly, sucrose and ethanol intakes were significantly lower in CB1-/- vs. CB1+/+ mice. In CB1 deficient mice, SR141716 had no effect in these models. PMID- 12770701 TI - Molecular biology and pathogenesis of Kaposi sarcoma-associated herpesvirus. AB - Kaposi sarcoma (KS)-associated herpesvirus (KSHV) is the most recently discovered human oncogenic herpesvirus. The virus is associated with KS lesions and other human malignancies, including pleural effusion lymphomas and multicentric castleman's disease. The sequence of the viral genome demonstrated that it belongs to the gammaherpesvirus family similar to the Epstein-Barr virus, the only other known human herpesvirus associated with human cancers. Molecular studies have identified a number of viral genes involved in regulation of cell proliferation, gene regulation, chromatin remodeling and apoptosis. KSHV transforms human endothelial cells in vitro with low efficiency and expresses a repertoire of latent genes involved in the establishment of latency. One of these latent proteins, the latency-associated nuclear antigen (LANA) is required for episomal maintenance and tethers the viral genome to the host chromatin. LANA has now been shown to be a multifunctional protein involved in numerous cellular functions including binding to the retinoblastoma protein and p53, regulating cell proliferation and apoptosis. PMID- 12770702 TI - Induction of Oenococcus oeni H+-ATPase activity and mRNA transcription under acidic conditions. AB - The profiles of Oenococcus oeni IOB84.13 H(+)-ATPase activity under various conditions of growth were studied. Cells growing at low pH 3.5 had a 1.6-fold higher H(+)-ATPase activity compared to control cells grown at pH 5.3. While the pH of the growth medium was shown to be stable in the presence of malic acid, a drastic decrease in pH from 5.3 down to 3.9 during growth in the absence of malic acid induced an increase in H(+)-ATPase activity by 1.5-fold. This induction was even greater when the initial pH was 3.5. Partial cloning of the genes encoding the beta-subunit and the epsilon-subunit of the H(+)-ATPase suggested a typical F(1)F(0)-ATPase genetic organization in O. oeni. The atp mRNA was detected by slot blots. Cells shocked at acidic pH were shown to contain higher levels of atp mRNA compared to the control cells grown at pH 5.3. Taken together, these results indicate that the H(+)-ATPase of O. oeni is induced at low pH and that regulation seems to occur at the level of transcription. This agrees with the role of this enzyme in the regulation of the cytoplasmic pH and in the acid tolerance of O. oeni. PMID- 12770703 TI - NaCl-sensitive mutant of Staphylococcus aureus has a Tn917-lacZ insertion in its ars operon. AB - Staphylococcus aureus is a Gram-positive bacterium that is extremely halotolerant. To investigate the molecular mechanisms by which S. aureus can cope with osmotic stress, Tn917-lacZ-induced NaCl-sensitive mutants were isolated. An NaCl-sensitive mutant showed a longer lag period, slower growth rate, and lower final culture turbidity than the parent strain in liquid medium containing 1.5 M NaCl. Electron microscopic observation of the NaCl-sensitive mutant under NaCl stress conditions revealed large, pseudo-multicellular cells. Addition of exogenous osmoprotectants, such as glycine betaine, choline, L-proline, and proline betaine, did not relieve the NaCl sensitivity of the mutant. The region flanking the transposon insertion site in the NaCl-sensitive S. aureus chromosome was sequenced. The mutated gene was 99% identical to arsR, the arsenic operon regulatory protein present on the pI258 plasmid of S. aureus. The ars operon from pI258 was subcloned into the shuttle vector pLI50 and transferred into the NaCl sensitive mutant. The ars operon in trans restored NaCl tolerance in the mutant, suggesting that NaCl sensitivity is due to the mutation in arsR. PMID- 12770704 TI - Purification and characterization of nitric oxide synthase from Staphylococcus aureus. AB - We previously reported the presence of nitric oxide synthase (NOS) in Staphylococcus aureus ATCC6538P whose activity was induced by methanol. In the present study, the methanol-induced NOS was purified 900-fold from S. aureus by means of Mono Q ion exchange column, 2',5'-ADP-agarose affinity column, and Superdex 200HR gel permeation column chromatography. The purified bacterial NOS showed two protein bands with 67 and 64 kDa molecular mass on SDS-PAGE. However, the molecular mass of the NOS was 135 kDa on Superdex 200HR gel permeation column chromatography, indicating that the native enzyme exists as a heterodimer. This bacterial NOS had K(m) value of 13.4x10(-6) M for L-arginine and V(max) of 35.3 nmol min(-1) mg(-1) protein. In addition, reduced nicotinamide adenine dinucleotide phosphate, flavin adenine dinucleotide, flavin mononucleotide, tetrahydrobiopterin, calmodulin and Ca(2+) were required as cofactors in the conversion of L-arginine to L-citrulline, and NOS inhibitors selectively inhibited the activity of the purified NOS. PMID- 12770705 TI - Microbial hydroxylation of progesterone with Acremonium strictum. AB - Microbial hydroxylation of progesterone occurred in the culture of Acremonium strictum PTCC 5282 to produce two hydroxylated pregnene-like steroids. The metabolites were purified and characterized using spectroscopic methods and identified as 15alpha-hydroxyprogesterone and 15alpha-hydroxydeoxycorticosterone. PMID- 12770706 TI - Distinguishing Ophiostoma ips and Ophiostoma montium, two bark beetle-associated sapstain fungi. AB - Two synonymous sapstain species, Ophiostoma montium and Ophiostoma ips, which are vectored by Dendroctonus ponderosae and various bark beetles, respectively, were differentiated into separate species using growth and molecular characteristics. Analysis of 32 isolates of the two species from different countries showed that O. ips was able to grow at 35 degrees C while O. montium was not. This growth based differentiation was strongly supported by sequence data for the internal transcribed spacer (ITS), 5.8S and partial 28S rDNA, and the beta-tubulin genes. The beta-tubulin gene sequence data indicate that the two species can easily be differentiated with a single polymerase chain reaction (PCR) assay. PMID- 12770707 TI - Molecular and biochemical analysis of the system regulating the lytic/lysogenic cycle in the pneumococcal temperate phage MM1. AB - The temperate phage MM1 forms stable lysogens in Streptococcus pneumoniae. We report here the first characterization of the lysogenic control region in Pneumococcus which contains two functional divergent promoters (P(R) and P(L)). MM1 encodes a 14-kDa cI protein (CI) that appears to be responsible for maintaining the lysogenic state in Pneumococcus since it prevents elongation of the transcripts controlled by P(R) and P(L). PMID- 12770708 TI - Multilocus sequence typing (MLST) of Escherichia coli O78 strains. AB - Strains of Escherichia coli serotype O78 are associated with many diseases, including invasive infections, in humans and farm animals. The clonal relationship between strains from different hosts is therefore important for assessing the risk of zoonotic infections. Here we propose a multilocus sequence typing scheme for E. coli, based on six housekeeping genes. Preliminary, but significant, results indicate that clonal division in E. coli O78 strains is host independent, and closely related clones reside in different hosts. There was a positive correlation between virulence and clonal origin. PMID- 12770709 TI - A metal-independent hydrolase from a Penicillium oxalicum strain able to use phosphonoacetic acid as the only phosphorus source. AB - A Penicillium oxalicum strain was capable of the phosphate-sensitive utilization of phosphonoacetic acid as the sole source of phosphorus. A carbon-to-phosphorus bond-cleavage enzyme yielding acetic acid and inorganic phosphate was detected and characterized in extracts from cells grown on this phosphonate. Contrary to bacterial phosphonoacetate hydrolases, the fungal enzyme neither required nor was stimulated by divalent cations. PMID- 12770710 TI - Regulation of biosynthesis and transport of aromatic amino acids in low-GC Gram positive bacteria. AB - Computational comparative techniques were applied to analysis of the aromatic amino acid regulon in Gram-positive bacteria. A new candidate transcription regulation signal of 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase and shikimate kinase genes was identified in Streptococcus and Lactococcus species. New T-boxes were found upstream of aromatic amino acid biosynthesis and transport genes in the Bacillus/Clostridium group. The substrate specificity of proteins from the PabA/TrpG family was assigned based on metabolic reconstruction and analysis of regulatory signals and phylogenetic patterns. New candidate tryptophan transporters were identified; their specificity was predicted by analysis of T-box regulatory sites. Comparison of all available genomes shows that regulation of genes of the aromatic amino acid biosynthesis pathway is quite labile and involves at least four regulatory systems, two at the DNA level and two more involving competition of alternative RNA secondary structures for transcription and/or translation regulation at the RNA level. PMID- 12770711 TI - Cloning and characterization of genes specifically expressed during infection stages in the rice blast fungus. AB - We isolated promoters of 12 genes from the rice blast fungus based on the sequences of randomly selected expressed sequence tags (ESTs) (appressorium formation stage cDNA library of Magnaporthe available from GenBank). These promoters (and the 5' coding regions if any) were fused in frame with egfp, and their expression patterns were examined under the epifluorescence microscope. Among them, two turned out to be specifically active in structures necessary for infection, viz. a promoter of adenylate cyclase interacting protein 1-like gene expressed in conidia, germ tubes, and appressoria, and a promoter of putative membrane-associated or secreted protein gene specifically expressed in appressoria. Although targeted knockout mutants of either gene failed to show detectable phenotypic alterations under laboratory conditions, these ESTs should be useful for identification of genes expressed during infection stages. PMID- 12770712 TI - Cloning, characterization and heterologous expression of the Blakeslea trispora gene encoding orotidine-5'-monophosphate decarboxylase. AB - The pyrG gene of the fungus Blakeslea trispora, encoding orotidine-5' monophosphate decarboxylase (OMPD) enzyme, was cloned by heterologous hybridization of a genomic library with the Mucor circinelloides pyrG gene. The deduced amino acid sequence of the B. trispora pyrG gene is highly similar to the OMPD from other organisms. Hybridization analyses revealed that the only copy of this gene present in the genome of B. trispora is constitutively expressed. Heterologous complementation of a mutant of M. circinelloides deficient in OMPD activity with the B. trispora pyrG gene and promoter sequence confirmed the function of this gene. This functional complementation demonstrates that heterologous expression in M. circinelloides might be used to investigate the function of genes of B. trispora. PMID- 12770713 TI - Expression and biochemical characterization of the 1-HO-carotenoid methylase CrtF from Rhodobacter capsulatus. AB - In purple bacteria, acyclic 1-methoxy carotenoids like spheroidene or spirilloxanthin are essential components of the photosynthetic apparatus. One of the last steps of their biosynthesis involves O-methylation of the 1-hydroxy group. The 1-HO-carotenoid methylase CrtF from Rhodobacter capsulatus catalyzing this type of reaction was expressed in Escherichia coli in an active form. It was then purified by affinity chromatography and biochemically characterized. The enzymatic reaction depends on S-adenosylmethionine as the only cofactor. By complementation in E. coli, the substrate specificity of the enzyme was determined. It could be shown that the enzyme converts not only all possible 1 hydroxy carotenoids in the spheroidene/1'-HO-spheroidene biosynthetic pathway of R. capsulatus but also carotenoid intermediates leading to the formation of spirilloxanthin in a pathway which is absent in R. capsulatus but present in related species. PMID- 12770714 TI - Characterization of nitrogen-fixing Paenibacillus species by polymerase chain reaction-restriction fragment length polymorphism analysis of part of genes encoding 16S rRNA and 23S rRNA and by multilocus enzyme electrophoresis. AB - Forty-two strains representing the eight recognized nitrogen-fixing Paenibacillus species and 12 non-identified strains were examined by restriction fragment length polymorphism (RFLP) analysis of part of 16S and 23S rRNA genes amplified by polymerase chain reaction (PCR). Eleven different 16S rDNA genotypes were obtained from the combined data of RFLP analysis with four endonucleases and they were in agreement with the established taxonomic classification. Only one group of unclassified strains (Group I) was assigned in a separate genotype, suggesting they belong to a new species. Using the 23S PCR-RFLP method only six genotypes were detected, showing that this method is less discriminative than the 16S PCR RFLP. Using the multilocus enzyme electrophoresis (MLEE) assay, the 48 strains tested could be classified into 35 zymovars. The seven enzymatic loci tested were polymorphic and the different profiles obtained among strains allowed the grouping of strains into 10 clusters. The PCR-RFLP methods together with the MLEE assay provide a rapid tool for the characterization and the establishment of the taxonomic position of isolates belonging to this nitrogen-fixing group, which shows a great potentiality in promoting plant growth. PMID- 12770715 TI - BphK shows dechlorination activity against 4-chlorobenzoate, an end product of bph-promoted degradation of PCBs. AB - A bphK gene encoding glutathione S-transferase (GST) activity is located in the bph operon in Burkholderia sp. strain LB400 but its role in polychlorinated biphenyl (PCB) metabolism is unknown. This gene was over-expressed in Escherichia coli and an in vivo assay based on growth of E. coli containing GST activity was used to identify potential novel substrates for this enzyme. Using this assay, 4 chlorobenzoate (4-CBA) was identified as a substrate for the BphK enzyme. High pressure liquid chromatography analysis and chloride ion detection showed removal of 4-CBA and an equivalent increase of chloride in cell extracts when incubated with this enzyme. These results would indicate that this BphK enzyme has dechlorination activity in relation to 4-CBA and may have a role in protection of other Bph enzymes against certain chlorinated metabolites of PCB degradation. PMID- 12770716 TI - Interactions of the Caulobacter crescentus XerC and XerD recombinases with the E. coli dif site. AB - In most bacteria, chromosome dimers arise from homologous recombination between replicated chromosomes. These dimers are then resolved by the action of the XerC and XerD recombinases, which act on the chromosomal dif site in the presence of the FtsK cell division protein. We have cloned the xerC and xerD genes from Caulobacter crescentus, and overexpressed them as maltose-binding protein fusion proteins. These fusion proteins were purified and used in in vitro DNA-binding assays to the Escherichia coli dif site with each protein individually, and in combination with each other. In addition, combinations of Xer proteins from E. coli were also tested for cooperativity with the corresponding C. crescentus proteins. PMID- 12770717 TI - Novel protein vaccine candidates against Group B streptococcal infection identified using alkaline phosphatase fusions. AB - Using an alkaline phosphatase-based genetic screening method, we identified a number of proteins that are potentially located on the outer surface of Group B streptococcus (Streptococcus agalactiae). In an enzyme-linked immunosorbent assay, antisera raised against two of the proteins, the streptococcal yutD homologue and a subunit of an ABC transporter, recognised clinically important serotypes of Group B streptococcus. In a neonatal rat model, purified IgG from the sera conferred significant levels of protection against a lethal challenge infection. The proteins identified show potential as protein subunit candidates for vaccines against Group B streptococcal disease in neonates. PMID- 12770718 TI - A method for allelic replacement in Francisella tularensis. AB - A vector for mutagenesis of Francisella tularensis was constructed based on the pUC19 plasmid. By inserting the sacB gene of Bacillus subtilis, oriT of plasmid RP4, and a chloramphenicol resistance gene of Shigella flexneri, a vector, pPV, was obtained that allowed specific mutagenesis. A protocol was developed that allowed introduction of the vector into the live vaccine strain, LVS, of F. tularensis by conjugation. As a proof of principle, we aimed to develop a specific mutant defective in expression of a 23-kDa protein (iglC) that we previously have shown to be prominently upregulated during intracellular growth of F. tularensis. A plasmid designated pPV-DeltaiglC was developed that contained only the regions flanking the encoding gene, iglC. By a double crossover event, the chromosomal iglC gene was deleted. However, the resulting strain, denoted DeltaiglC1, still had an intact iglC gene. Southern blot analysis verified that LVS harbors two copies for the iglC gene. The mutagenesis was therefore repeated and a mutant defective in both iglC alleles, designated DeltaiglC1+2, was obtained. The DeltaiglC1+2 strain, in contrast to DeltaiglC1, was shown to display impaired intracellular macrophage growth and to be attenuated for virulence in mice. The developed genetic system has the potential to provide a tool to elucidate virulence mechanisms of F. tularensis and the specific F. tularensis mutant illustrates the critical role of the 23-kDa protein, iglC, for the virulence of F. tularensis LVS. PMID- 12770719 TI - Regulation of hyphal growth and sporulation of the insect pathogenic fungus Entomophthora thripidum in vitro. AB - Entomophthora thripidum is an obligate biotrophic insect pathogenic fungus that grows as protoplasts within the hemocoel of thrips. Prior to penetration through the insect cuticle and spore formation at the insect surface the protoplasts switch to hyphal growth. In vitro, the differentiation to hyphal growth was a prerequisite for the subsequent formation of infectious spores and was detected 10-20 days after inoculation. E. thripidum secreted a factor that autoinduced the differentiation to hyphal growth. The discovery of this activity inducing hyphal growth made possible the reliable production of spores, the infection of host insects and the consecutive re-isolation of the fungus from the infected insects. PMID- 12770720 TI - The IS1 elements in Shigella boydii: horizontal transfer, vertical inactivation and target duplication. AB - IS1(SB) and its two variants were identified as the major and minor IS1 elements in Shigella boydii. The nucleotide sequences of IS1(SB), IS1(O157:H7) from Escherichia coli O157:H7 and IS1F from E. coli K12 suggest that these IS1 elements had been horizontally transferred among S. boydii and E. coli O157:H7 and K12. The two IS1(SB) variants and IS1(O157:H7) have transposition activities 7- to 86-fold less than that of IS1(SB), whereas IS1F has little transposition activity. Analysis of the flanking sequences of IS1(SB) and its two variants in S. boydii revealed the nature of regional specificity of the target sites and the sequence dependence of 8 and 9 bp target duplications, for which a model is presented. PMID- 12770722 TI - The aging gonad. PMID- 12770723 TI - H1oo: a pre-embryonic H1 linker histone in search of a function. AB - The mouse oocyte-specific linker histone H1oo (1) constitutes a novel mammalian homologue of the oocyte-specific linker histone B4 of the frog and of the cs-H1 linker histone of the sea urchin; (2) is expressed as early as the germinal vesicle (PI) stage oocyte, persisting into the MII stage oocyte, the oocytic polar bodies, and the 2-cell embryo, extinction becoming apparent at the 4-8 cell embryonic stage; and (3) may play a key role in the control of gene expression during oogenesis and early embryogenesis, presumably through the perturbation of chromatin structure. PMID- 12770724 TI - Regulation of the phenotype of ovarian somatic cells by estrogen. AB - The pathway of mammalian sex determination, and subsequent differentiation of the gonads is under the control of the sex-determining gene, Sry, on the Y chromosome. The presence of Sry leads to the formation of a testis with its complement of Sertoli and Leydig somatic cells. In the absence of Sry, an ovary develops with granulosa and theca cells. Ovarian development is said to initiate in the XX gonad as a default pathway because the XX cells do not express Sry. This review summarizes evidence supporting the view that the ovary is not entirely a default gonad. Studies of mice with deletions in both estrogen receptor (ERalphabetaKO) or aromatase (ArKO) genes have identified an important role of estrogens in maintaining differentiation and development of somatic cells in the ovary of eutherian mammals. In the absence of estrogen (ArKO) or the capacity to transduce an estrogen signal (ERalphabetaKO), the somatic cells in the ovary exhibited a male phenotype including Sertoli and Leydig cells. When ArKO mice were replaced with estrogen, the male phenotype was diminished and there was evidence of normal folliculogenesis in the ovary. It is concluded that the differentiation of somatic cells in the eutherian ovary is influenced by the sex steroid environment. PMID- 12770725 TI - Production of vascular endothelial growth factor and angiogenic factor in human follicular fluid. AB - The aim of this study was to quantitate of vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), and angiogenin in follicular fluid (FF) and to correlate the levels of these substances with oocyte maturation. FF were aspirated from women undergoing in vitro fertilization and embryo transfer. Sera were collected from women with normal menstrual cycles. VEGF in FFs and sera were measured by enzyme linked immunosorbent assay. VEGF, HGF, and angiogenin mRNA expression aspirated folliculars cell was analyzed by reverse transcription and polymerase chain reaction (RT-PCR). The concentrations of VEGF, HGF, and angiogenin in FF were significantly higher than those in serum (P<0.001). VEGF, HGF, and angiogenin mRNA in the aspirated follicles cell was detected by RT-PCR. HGF levels were higher in FF containing mature oocyte. The levels of VEGF in FF containing mature oocytes in women under 39 years of age were significantly lower than those in FF from women more than 40 years old (P<0.01). Our data suggest that VEGF, HGF, and angiogenin may play an important role in follicular growth and development, that VEGF levels in FF appear to be age-dependent; and that VEGF and HGF levels might be valuable biochemical markers of oocyte maturation. PMID- 12770727 TI - Growth differentiation factor-9 signaling in the ovary. AB - Growth differentiation factor-9 (GDF-9) is an oocyte-derived growth factor and a member of the transforming growth factor-beta (TGF-beta) superfamily. In GDF-9 null mice, follicle development is arrested at the primary stage and in vivo treatment with GDF-9 enhances the progression of primordial and primary follicles into small preantral follicles. In vitro, GDF-9 promotes granulosa cell proliferation but inhibits FSH-induced differentiation. GDF-9 also promotes the differentiation of theca cells in vivo and in vitro. GDF-9, like TGF-beta or activin, is a close member of the bone morphogenetic proteins (BMPs) family. GDF 9 likely initiates signaling by assembling two related but distinct types of receptors, both of which are serine/threonine kinases with a single transmembrane domain. The ligand-receptor binding activates intracellular transcription factors called Smads. In granulosa cells, Vitt et al. have shown that the BMP receptor type II is involved in GDF-9 signaling. The type I receptors and the Smad pathway for GDF-9 remain to be identified. PMID- 12770726 TI - Regulation of NGFI-B expression during the ovulatory process. AB - NGFI-B is an immediate-early gene that encodes an orphan nuclear receptor. In the rat ovary, the preovulatory surge of LH induces NGFI-B expression in granulosa cells of preovulatory follicles, reaching a peak within 1 h and declining to control levels at 6 h. The LH-stimulated NGFI-B expression is abolished by alpha amanitin, but superinduced by cycloheximide. Similarly, treatment of human luteinized granulosa cells with LH causes a rapid and transient stimulation of NGFI-B expression. Interestingly, the induction of NGFI-B expression in response to LH stimulation in preovulatory granulosa cells requires signaling through protein kinase Czeta. Furthermore, two other NGFI-B family members, Nurr1 and Nor1, are also rapidly stimulated by LH in granulosa cells of preovulatory follicles through the activation of protein kinase Czeta. The cell-type specific expression and LH induction of NGFI-B suggests a potential role of NGFI-B in the ovulatory process. PMID- 12770728 TI - What we have learned from isolated cells from human ovary? AB - In the ovary, morphodynamics of follicles with cyclic maturation, ovulation and repair occur under the control of various tropic factors. The ovarian functions have been mostly studied by using subhuman primates and non-primate animals because of the limited availability of closely staged human specimens. We have recently established the in vitro culture systems of ovarian surface epithelium (OSE) and granulosa cells of humans, and subsequently developed the immortalization of each cell. The immortalized cell lines may supply us advanced studies on ovarian disorders as well as its physiological functions. On the embryologically putative mullerian potential of coelomic epithelium, endometriosis can be explained by coelomic metaplasia from the peritoneal mesothelium, including OSE. We can microscopically observe a continuity from flat epithelial cells on the ovarian surface or within the cortical inclusion cysts to endometriotic gland cells. The primary human OSE cells exhibited a glandular stromal structure similar to endometriosis when they were co-cultured with endometrial stromal cells in an estrogen-rich environment. Primary and immortalized OSE cells converted the estrone to estradiol, and expressed the genes for steroidogenic factor-1 (SF-1), p450arom and 17beta-HSDs. This character of OSE was, in part, similar to the granulosa cells. One of the immortalized OSE clone produces disseminated tumors mimicking undifferentiated carcinomas in nude mice. Ovarian granulosa cells play a key role in the functional maturation of the entire follicle. The molecular pathways in granulosa cells responsible for the growth, differentiation, and nursing the oocyte are still largely unknown. Our immortalized human granulosa cell line, GC1a, obtained from developing follicles, showed no steroid hormone biosynthesis, and no detectable expression of the genes for StAR or cytochrome p450 enzymes due to the lack of SF-1. Transfected SF-1 elicited estradiol secretion in GC1a cells with concomitant expression of the genes encoding the proteins for gonadal steroidogenesis. The enzymatic activity of 17beta-HSD was also achieved by SF-1 transgene. These results indicate that SF 1 controls the gene expression required for steroidogenesis in the human developing follicle. Clinically, immortalized GC1a cells from human origin, with steroidogenic capacity, may serve as a feeder layer for in vitro oocyte maturation. Further investigations of our immortalized OSE and granulosa cells of humans will allow us to clarify whether they have a single progenitor cell. PMID- 12770729 TI - Ovarian hyperstimulation syndrome-model rats; the manifestation and clinical implication. AB - Ovarian hyperstimulation syndrome (OHSS) is the most serious complication of ovulation induction with the gonadotropins. Though vascular endothelial growth factor (VEGF) has been implicated as a prime causative factor of OHSS progression, other factors must also be involved in the pathogenesis of OHSS. We have established an experimental model of OHSS in immature female rats. The ovarian weights and vascular permeability of the OHSS model rats are significantly increased by the ovarian stimulation with human chorionic gonadotropin (hCG). hCG elicits VEGF production in OHSS model rat ovaries. The addition of potent synthetic progesterone antagonist RU486, which reduces the extension of OHSS, attenuates the ovarian kinin and VEGF production dose dependently whereas the VEGF gene expression was stable. VEGF protein detected in both the lung and the liver of the OHSS model rats is not affected at 24 h after the addition of hCG and RU486 in contrast to the ovary. Our results demonstrate that progesterone is implicated in the development of OHSS, in part, to enhance the ovarian VEGF production by post-transcriptional and organ-specific control. PMID- 12770730 TI - Ovarian follicle populations of the rat express TGF-beta signalling pathways. AB - The transforming growth factor-beta (TGF-beta) superfamily comprises more than 40 members, classified on the basis of structural similarity. These factors elicit a diverse range of cellular responses in insects, nematodes and vertebrates, via serine/threonine kinase receptors and intracellular Smad proteins, which when activated mediate gene transcription. Some members of the superfamily, notably activin, TGF-beta, GDF-9 and the bone morphogenetic proteins have been shown to influence ovarian function. Despite these actions, TGF-beta superfamily signalling pathways and specifically those within follicle population subtypes, have been poorly characterised in the ovary. We have shown that the ovary contains type I and II receptors and Smads, which enable it to transduce signals in response to TGF-beta superfamily members. It remains to be established however, as to which follicle subtypes these pathways are active in. PMID- 12770731 TI - START domain proteins and the intracellular trafficking of cholesterol in steroidogenic cells. AB - The intracellular trafficking of cholesterol in steroidogenic cells plays an important role in the regulation of hormone synthesis. Recent evidence indicates that a family of proteins related to the steroidogenic acute regulatory protein (StAR) perform critical functions in moving the sterol substrate to the mitochondrial inner membrane where the first committed step in steroid hormone synthesis occurs. StAR, the prototype of the family, is known to promote the translocation of cholesterol from the outer to the inner mitochondrial membrane. Mutations in StAR cause congenital lipoid adrenal hyperplasia, a cholesterol storage disorder in which synthesis of all gonadal and adrenocortical steroid hormones is severely impaired, and the cholesterol that is not efficiently moved into the mitochondria accumulates in cytoplasmic lipid droplets. The StAR-related lipid transfer (START) domain consists of an approximately 210 amino acid residue sequence that forms a compact alpha/beta structure, a helix-grip fold, with a hydrophobic tunnel that can accommodate a sterol molecule. START domains can bind sterol, facilitate the transfer of cholesterol from sterol-rich unilammelar liposomes to acceptor membranes, and stimulate steroidogenesis when expressed in cells co-expressing the cholesterol side-chain cleavage system or when added to isolated steroidogenic mitochondria. Sixteen human START domain proteins have been identified to date. Of these, StAR and MLN64 consist of one subfamily and newly described proteins named StarD4, StarD5, and StarD6 represent a closely related second subfamily. MLN64 is incorporated into the late endosomal compartment and is involved in the movement of cholesterol acquired from endocytosed LDL out of these vesicles. Expression of a dominant negative form of MLN64 causes accumulation of free cholesterol in lysosomes. The roles of StarD4, StarD5, and StarD6 in sterol movement remain to be determined. These genes have tissue-specific patterns of expression that may predict specialized roles. PMID- 12770732 TI - The generation and characterization of an ovary-selective cDNA library. AB - We have utilized the differential screening technique suppression subtractive hybridization to systematically isolate and clone genes that are expressed in an ovary-selective/specific manner. In total, 844 clones were sequenced and analyzed for homology to known genes using the basic local alignment tool. One hundred and fifty nine independent clones proved identical to previously-characterized genes whereas an additional 100 independent clones proved significantly homologous (but not identical) to previously-characterized genes. Yet 83 other independent clones did not display significant homology to previously-characterized genes now listed in the publicly-accessible non-redundant databases. As such, these latter genes were deemed novel. In this communication we focus on two such novel ovary specific/hormonally-dependent genes, the full-length sequences of which were isolated using RACE technology. These ovary-selective genes may have significant implications for the understanding of ovarian function in molecular terms and for the development of innovative strategies for the promotion of fertility or its control. PMID- 12770733 TI - Hemodynamic response of ovarian artery after hCG injection. AB - We have analyzed ovarian hemodynamics immediately after human chorionic gonadotropin (hCG) administration in patients treated by clomiphene-hCG and human menopausal gonadotropin-hCG. This study involved 40 infertile women who signed consents to participate in this study. After intramuscular injection of 10000 IU hCG, the change of ovarian arterial blood flow (BF) was evaluated by color Doppler. Pulsatility index, resistance index, maximum velocity (V(max)), mean velocity, minimum velocity, cross-sectional area of ovarian artery (Area) and BF were measured before and 15-180 min after hCG administration. In the 36 subjects in which ovulation was induced successfully, V(max) and BF increased significantly even at 15 min after hCG administration and thereafter. In the 4 non-ovulatory subjects, no significant changes in any of indices at any of measured time points were observed. Comparative study of non-ovulatory and ovulatory subjects suggested that ovulation may be predicted by the ovarian hemodynamic analysis immediately after hCG administration. PMID- 12770734 TI - Analysis of signal transduction stimulated by gonadotropins in granulosa cells. AB - Gonadotropins exert their effect on ovarian follicular cells through the activation of the hormone sensitive adenylate cyclase and consequent elevation of intracellular cyclic AMP (cAMP). Desensitization to the hormone in cultured primary granulosa cells can occur within a short period and internalization of the hormone-receptor complex has been observed both in vivo and in vitro. It was recently documented that the gonadotropins as well as cAMP activate MAP kinase (MAPK) in granulosa cells. Moreover we discovered that specific inhibitors of extracellular signal-regulated kinase phosphorylation, 1 and 2, augment steroidogenesis in granulosa cells up-regulating steroidogenic acute regulatory (StAR) protein expression, and that this modulation is blocked by specific inhibitors of protein kinase A. It is therefore suggested that gonadotropins may activate both stimulatory and inhibitory pathways which regulate steroidogenesis. Moreover the ratio between the activity of these two pathways may determine the rate of steroidogenesis, and rapid activation of MAPK may account as part of the mechanism of desensitization to the hormonal action. Steroidogenic factor-1 and DAX-1 may be involved in the regulation of the MAPK-dependent attenuation of steroidogenesis, since they exhibit sites that could be potentially phosphorylated by the MAPK cascade. PMID- 12770735 TI - Hepatocyte growth factor modulates motility and invasiveness of ovarian carcinomas via ras mediated pathway. AB - Hepatocyte growth factor (HGF) is a multifunctional growth factor which has pleiotrophic biological effects on epitherial cells, such as proliferation, motogenesis, invasiveness and morphogenesis. Peritoneal dissemination is critical for the progression of ovarian cancer and our study revealed that HGF induces migration and invasion of ovarian cancer cells. We also demonstrated that HGF stimulates autophosphorylation of its receptor, followed by activation of the Ras MAP (mitogen-activated peptide) kinase cascade. Moreover, infection of ovarian cancer cells with Ras dominant-negative adenovirus reduced the HGF-induced motogenic and invasive activities. Additionally, both MEK and PI3-kinase pathways downstream of Ras was involved in HGF-stimulated ovarian cancer cell invasiveness. PMID- 12770736 TI - Cell-cell adhesion in the normal ovary and ovarian tumors of epithelial origin; an exception to the rule. AB - Cells are held together either by direct cell-cell contact or adhesion to extra cellular matrix. Cell-cell adhesion in epithelial cell sheets consists of junctions, i.e. tight-, adherens- and gap-junctions. The adherens junctions, which are build up by the cadherin/catenin complex, are the main topic of this review, especially the aspect of its role in ovarian tumor biology. The ovarian surface epithelium is the origin for approximately 90% of the malignant ovarian tumors. The tumors arise from the inclusion cysts, localized in the ovarian stroma and grow solid, cystic or in mixed formations. Intra-abdominal spread of the ovarian cancer is common and this is a process that theoretically could be closely connected with impaired cell-cell adhesion. However, as we stand today, descriptive and functional studies on the cadherin-catenin complex and its cell signaling role in ovarian tumorigenesis reveals data that suggests a conversion of the mesothelial-like cells of the ovarian surface to a more epithelial phenotype with normal cell-cell adhesion prior to tumor differentiation. In later stages, invasive ovarian tumors still strongly express several cadherins, which are contrary to many other tumors, i.e. prostate and thyroid adenocarcinomas. PMID- 12770737 TI - Genetic alterations in ovarian carcinoma: with specific reference to histological subtypes. AB - Multiple genetic changes including activation of proto-oncogenes and inactivation of tumor suppressor gene are involved in the development of human ovarian cancer. We describe such genetic alterations with specific reference to histological subtypes. K-ras activation is specific for mucinous tumors including adenomas. Borderline tumors and carcinomas, suggesting that K-ras activation may be associated with the mucinous differentiation rather than malignant transformation. Inactivation of p53 is detected in 30-40% of ovarian carcinoma. Mutations are more frequently observed in serous carcinomas, but not found in adenomas or rarely found in borderline tumors, suggesting that p53 mutations may be directly involved in malignant transformation. TGFbeta-2 mutations are found in 50% of endometrioid carcinoma, but rarely in other type. Loss of DCC mRNA expression is found in 50% of serous carcinomas but less frequently in other type. Loss of DCC expression is rare in borderline tumors and adenomas, suggesting that inactivation of DCC may be directly involved in malignant transformation. Microsatellite instability (MI) is found in 17% of ovarian carcinomas, and is frequently found in endometrioid carcinomas. Although inactivation of p16 by point mutation or deletion is rare, p16 inactivation by loss of expression is relatively common. PMID- 12770738 TI - Effects of cryopreservation on meiotic spindles of oocytes and its dynamics after thawing: clinical implications in oocyte freezing--a review article. AB - Embryo freezing has been a successful practice, but oocyte cryopreservation formerly achieved poorer results. This was mainly due to low rates of survival, fertilization, and development. The major dissimilarities for oocytes to embryos are the character of the plasma membrane, the presence of cortical granules, at the metaphase of meiosis II with the spindle system. In addition, the oocytes must be fertilized by sperm at the appropriate time. To improve the survival rate, a refined slow freezing method with increased sucrose concentration would dehydrate oocytes more sufficiently. Vitrification is another approach to prevent ice crystal formation. Intracytoplasmic sperm injection is used to overcome possible zona hardening from the release of cortical granules. The microtubules of meiotic spindles are vulnerable to the thermal changes and would depolymerize. Cryopreserved oocytes exhibited serious disturbances of the microtubules immediately after thawing. Fertilization of oocytes with disorganized spindles could lead to chromosomal aneuploidy, digyny, and arrest of cleavage. After incubation, the microtubules would repolymerize in a time-dependent way. Normal fertilization and development of cryopreserved oocytes improved after appropriate incubation and timing of insemination, compatible with recovery of the spindles. With the improvement of survival, fertilization, and cleavage, oocyte cryopreservation would gain an imperative role. PMID- 12770739 TI - Estrogen receptor alpha polymorphisms and renal cell carcinoma--a possible risk. AB - Renal cell carcinoma is the most common form of kidney cancer. However, the genetic basis of renal cancer is not fully understood. Estrogens and their receptors (ERs) have been shown to play a role in various cancers and it is speculated that they can also affect the human kidney. One of the animal models utilized to study the effects of estrogens on renal cancer is the Syrian hamster. Exposing these hamsters to estrogens results in the development of kidney cancer and thus, the hormone-ER complex may be playing a role. The ER is expressed in reproductive as well as non-reproductive tissues and is implicated in the control of proliferation, differentiation, and development of many tissues. There are two types of ERs and they are the alpha and beta forms. Genetic polymorphisms of various factors have been shown to play a role in the alteration of their functions. The NH2-terminal region of the ERalpha protein influences its structure and function and thus, inherited variants of the ERalpha gene may alter tissue responsiveness to estrogens and possibly lead to renal carcinogenesis. Polymorphisms have been determined in the coding region of the human ERalpha gene and are located at the following codons: 10 T-->C, 85 G-->C, 87 G-->C, 243 C-->T, 325 C-->G, and 594 G-->A. There are also two polymorphisms that have been identified in intron 1 and give rise to a PvuII and XbaI restriction site. These polymorphisms of ERalpha have been shown to be associated with various cancers. Based on the evidence, it is hypothesized that polymorphisms of the ERalpha gene are associated with renal cell carcinoma. PMID- 12770740 TI - Reproductive function after treatment of malignant germ cell ovarian tumors. AB - The outcome and reproductive function were examined among patients with malignant ovarian germ cell tumors treated since 1980. Between 1980 and 2001, fertility sparing surgery was performed in 26 women, 23 of whom received adjuvant chemotherapy. With a median follow-up of 66.6 months, all patients have been alive, with histological types of 6 immature teratomas, 8 dysgerminomas, 6 yolk sac tumors, and 6 mixed types. Clinical stages were involved of 17 early stage and 9 advanced stage patients. After treatment, 20 women out of 26 recovered menstruation within 6 months. During follow-up, two chemotherapy-untreated and one treated patients experienced 4 conceptions in total. A treated patient conceived but selected artificial termination by affection of chemotherapy. Conservative surgery with adjuvant chemotherapy is the standard approach to treat patients with malignant ovarian germ cell tumors. In these 20 years, we experienced no delivery, so that fertility seems to be seriously affected by treatments. PMID- 12770741 TI - Effect of IGF-1 and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on the expression of LH receptors during cell differentiation in cultured granulosa cells. AB - Ovarian granulosa cells undergo a complex differentiation process during the growth and maturation of ovarian follicle. This process includes the acquisition of cell surface LH receptor, which mediates the granulosa cell's ability to respond to circulating LH. The results of the actions of LH on the mature granulosa cell include steroidogenesis, luteinization, and ovulation. As such, induction of the LH receptor in granulosa cells is a critical step in reproductive physiology. In the present study, we attempted to assess the effects of IGF-1 and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on FSH-induced LH receptor expression in rat granulosa cells to understand the actions of these factors on normal reproductive function. Treatment with FSH, as expected, produced a substantial increase in LH receptor mRNA level, and concurrent treatment with increasing concentrations of IGF-1 brought about dose-dependent increases in FSH-induced LH receptor mRNA. On the other hand, the concurrent treatment of TCDD (10 pM) resulted in a significant decrease in LH receptor after 24 h. The decay curves for LH receptor mRNA transcript showed a significant increase in the half-life after the addition of IGF-1 and a significant decrease after addition of TCDD. These data suggests a possible role for changes in LH receptor mRNA stability in the IGF-1 and TCDD induced regulation of LH receptor in rat granulosa cells. The rates of LH receptor mRNA gene transcription, assessed by nuclear run-on transcription assay, were not increased by the addition of IGF-1, but decreased by the addition of TCDD. The data of IGF-1 present that the interface between circulating hormones and paracrine/autocrine systems could provide an important mechanism to amplify the effects of gonadotropin hormones at the local level. In addition, the endocrine-disrupting effects of TCDD are, at least in part, caused by direct action on the expression of LH receptor expression in granulosa cells. PMID- 12770742 TI - Novel genes regulated by gonadotropins in granulosa cells: new perspectives on their physiological functions. AB - Follicular stimulating hormone (FSH) is a key hormone secreted from the pituitary, which controls the development of the follicle-enclosed oocytes in the mammalian ovary by interacting with specific receptors located exclusively on granulosa cells. Its biological activity involves stimulation of intercellular communication and upregulation of steroidogenesis, yet the entire spectrum of genes which are regulated by FSH are not fully characterized. We have established rat and human FSH responsive granulosa cell lines, which express FSH receptors at 20-times higher rates compared to primary cells. Since the lines are monoclonal, they are expected to have a homogeneous composition of RNA among the entire cell population, which increases the probability of yielding a distinct view of genes modulated by FSH eliminating the possibility of other cell types contamination. Using Affymetrix DNA microarrays to uncover novel FSH-regulated genes, we discovered genes not reported earlier to be regulated by FSH. These include genes coding for (1) proteases; (2) growth factors and cytokines; (3) proteins involved in intercellular communication and connection with the nervous system; (4) protein phosphatases and kinases; (5) anti oxidants and anti-toxicants; (6) G coupled proteins. These findings can deepen our understanding in the mechanism of FSH action in stimulation of the development of the ovarian follicular cells, in the modulation of ovarian intracellular and intercellular communication and in the process of selection of the dominant follicle. When human granulosa cells, obtained from in vitro fertilization patients were exposed to either hLH- or hFSH stimulation and mRNAs of these cells were analyzed by DNA microarrays, novel genes, similar to those found modulated by FSH in FSH responsive cell lines, were discovered in the human primary cells. This suggests that the immortalized cell systems established in our laboratory could serve as a useful system expanding the spectrum of authentic genes modulated by gonadotropin stimulation in normal ovarian function and in ovarian malfunction. PMID- 12770743 TI - Activation of the luteinizing hormone receptor in the extracellular domain. AB - Glycoprotein hormone receptors have ligand-binding ectodomains consisting of leucine-rich repeats, followed by a conserved cysteine-rich hinge region to the seven transmembrane (TM) region. Based on constitutively active mutations at Ser 281 in the hinge region of the thyroid-stimulating hormone receptor, we mutated the conserved serine in the luteinizing hormone (LH) receptor (S277I) and observed increased basal cAMP production and ligand affinity by mutant receptor. Conversion of Ser-277 to all natural amino acids led to varying degrees of receptor activation. Hydropathy index analysis indicated that substitution of neutral serine with selective nonpolar hydrophobic residues (Leu>Val>Met>Ile) confers constitutive receptor activation. Furthermore, mutation of the angular proline near Ser-273 to flexible Gly also led to receptor activation. The findings suggest the ectodomain of LH receptor constrains the TM region. Point mutations in the hinge region or ligand binding could cause conformational changes in the TM region that result in Gs activation. PMID- 12770744 TI - Multi-factorial role of GnRH-I and GnRH-II in the human ovary. AB - Normal ovarian functions are regulated by a wide variety of endocrine hormones, local paracrine and autocrine factors, which functionally interact with each other in a highly coordinated fashion. Recent findings have demonstrated that both forms of gonadotropin-releasing hormone (GnRH-I and GnRH-II) are expressed in various compartments of the human ovary including the granulosa-luteal cells, ovarian surface epithelial cells and ovarian tumors, and their expressions have been shown to be tightly regulated by gonadal steroids and gonadotropins. Functionally, these neuropeptides exert diverse biological effects in the ovary via binding to their cognate receptors, supporting the notion that these peptides act as paracrine and autocrine factors in modulating local ovarian functions. In this review, we will summarize recent literatures regarding the regulation of GnRH-I and GnRH-II gene expressions in the human ovary, and discuss the possible signal transduction mechanisms by which these hormones exert their actions in the gonad. Recent cloning of the second form of the GnRH receptor (GnRH-II receptor) in primates and other vertebrates demonstrated that it was structurally, and thus, functionally distinct from the GnRH-I receptor. Cell proliferation studies showed that GnRH-II inhibited the growth of human ovarian cancer cells that express GnRH-II but not GnRH-I receptor, indicating that the GnRH-II binding sites are functional in these cells. However, it remains unknown if GnRH-II receptor is expressed as a full-length, properly processed and functional gene transcript in humans, and its potential physiological roles such as differential regulation of gonadotropin secretion, neuroendocrine modulation and female sexual behavior await further investigation. PMID- 12770745 TI - GnRH agonist action on granulosa cells at varying follicular stages. AB - In the present review, we attempt to summarize our recent research related to comparative effects of gonadotropin-releasing hormone (GnRH) agonist on the proliferation, apoptosis and differentiated function of cultured porcine granulosa cells from varying follicular stages. The inhibitory effect of GnRH agonist on Proliferating cell nuclear antigen positive rate of cultured cells was prominent in granulosa cells from small and medium but not from large follicles. By contrast, the inhibitory effect of GnRH agonist on 17beta-estradiol and progesterone secretion by cultured cells was prominent in granulosa cells from large but not from small and medium follicles. The stimulatory effect of GnRH agonist on apoptosis positive rate of cultured cells was, however, uniform regardless of the stages of follicular growth. These results demonstrate that GnRH agonist exerts diverse actions on granulosa cells over the course of follicular growth. One downregulates granulosa cell proliferation in immature follicles as well as steroidogenesis in mature follicles, and the other upregulates apoptosis of granulosa cells regardless of the stages of follicular growth. PMID- 12770746 TI - Relaxin signaling in reproductive tissues. AB - The insulin/relaxin peptide family includes insulin, IGFs, relaxin1-3, INSL3/RLF, INSL4, INSL5/RIF2 and INSL6/RIF1, many without functional characterization. Based on analysis of transgenic phenotypes and phylogenetic profiling, we have discovered that two orphan leucine-rich repeat-containing G protein-coupled receptors, LGR7 and LGR8, are cognate receptors for relaxin whereas INSL3 is a specific ligand for LGR8. With the identification of the relaxin receptors, it is now possible to investigate specific cells and tissues that are responsive to relaxin in diverse physiological and pathological conditions as well as to develop agonists and antagonists for LGR7 and LGR8 as therapeutics to treat different labor disorders. Furthermore, future functional characterization of the specificity of these pluripoentent receptors with peptide ligands could lead to the understanding of related orphan ligands and receptors. PMID- 12770747 TI - CYP1B1 gene in endometrial cancer. AB - Metabolic activation of estradiol has been shown to be a key factor in endometrial carcinogenesis. 4-hydroxy estrogens (CYP1B1 metabolites) received particular attention because of their causative role in malignant transformation of various organs including endometrium. CYP1B1 displays the highest level of expression in endometrium. 4-hydroxy estrogens can bind to DNA via their quinone metabolites and cause oxidative damage in endometrial cancer. Moreover, the 4 hydroxy estrogens bind to the estrogen receptor and have estrogenic effects on target tissues. Six polymorphisms of the CYP1B1 gene have been described of which four result in amino acid substitutions; 1-13C-->T, codon 48C-->G, codon 119G- >T, codon 432C-->G, codon 449T-->C and codon 453A-->G. The polymorphisms on exons 2 and 3 have significant effects on the catalytic function of CYP1B1. Polymorphisms on specific regions of CYP1B1 gene result in hyperactivation of the protein and can lead to a higher susceptibility in the incidence of various cancers. Thus, inherited alterations in CYP1B1 hydroxylation activity may be associated with significant changes in estrogen metabolism and, thereby, may possibly explain inter-individual differences in endometrial cancer risk associated with estrogen-mediated carcinogenesis. PMID- 12770748 TI - Transplantation of the uterus. AB - Most women with uterine factor infertility have today no prospect of carrying a pregnancy to term. The development of a method for transplantation of the human uterus would be a means for many of these women to become both genetic and gestational mothers. In this article we review the literature concerning the history and recent development in the area of uterine transplantation. We describe our newly developed model for heterotopic uterine transplantation in the mouse, which we are using for studies of pregnancy outcome and rejection mechanisms. We also address some of the specific questions that need to be solved before attempts to transplant the human uterus should be performed. PMID- 12770749 TI - The role of leptin during the development of mouse preimplantation embryos. AB - Leptin is known to regulate diverse reproductive functions, and recent studies have implicated involvement of leptin in the early mouse embryo development. The aim of the present study was to investigate the expression of leptin and its functional receptor (OB-Rb) in mouse oocyte and preimplantation embryo, and to examine whether leptin influenced the early embryo development. Leptin mRNA was detected in blastocyst and hatched blastocyst, and OB-Rb mRNA was detected in oocytes, 1-cell, 2-cell, morula, blastocyst and hatched blastocyst. As for the origin of leptin, leptin mRNA was identified in both the oviduct and uterus of the pregnant mouse. Furthermore, in the pregnant mouse, the levels of leptin in uterine fluid were higher than those in the non-pregnant mouse. Supplementation of culture medium with leptin promotes the development of preimplantation embryos from 2-cell stage to the blastocysts, fully expanded blastocysts and hatched blastocysts. Leptin significantly increased the total cell number of blastocysts, and the effect was preferentially observed in the trophectoderm. These findings raise the possibility that leptin regulates the development of mouse preimplantation embryo through a paracrine pathway. PMID- 12770750 TI - Expression of LDL receptor and uptake of LDL in mouse preimplantation embryos. AB - OBJECTIVES: The aim of this study is to address the role of low-density lipoprotein (LDL) on mouse preimplantation embryos. METHODS: The temporal expression of low-density lipoprotein receptor (LDLR) and side-chain cleavage cytochrome P450 (P450scc) mRNAs in mouse oocytes and preimplantation embryos up to the hatched blastocyst stage were analyzed by RT-PCR and nested PCR techniques. Simultaneously, the expression of LDLR in the protein level was analyzed by fluorescent immunohistochemistory at oocyte, 4-cell embryo and blastocyst. Uptake of LDL was analyzed using the LDL labeled with the fluorescent probe DiO. RESULTS: LDLR mRNA was detected at oocyte, 8-cell, morula, blastocyst and hatched blastocyst stages. P450scc mRNA was detected at oocyte, 1-cell, 2 cell, 4-cell, blastocyst and hatched blastocyst stages. LDLR protein was detected in blastocyst. P450scc protein was detected in oocyte, 4-cell, and blastocyst. LDL-DiO was taken into embryo at blastocyst stage and this uptake was competitively suppressed by excess unlabeled LDL. CONCLUSIONS: It is suggested from the present study that LDLR may play an essential role in uptake of exogenous LDL into blastocyst stage and cholesterol derived from LDL may be the source of steroid hormone synthesis. PMID- 12770751 TI - Immunological and endocrinological studies on lymphocyte subpopulation and medical treatment for infertility in patients with endometriosis. AB - We investigated the relationship between interleukin-6 (IL-6) levels and subset profiles of T lymphocyte (T-cell) and macrophage in peritoneal fluid (PF) with or without endometriosis (EM). IL-6 levels in PF with EM were significantly higher than those without EM. IL-6 producing cells with EM were analyzed in each activated mature T-cell (CD3+CD69+) and macrophage (CD14+) were 0.5 and 3.5%, respectively, whereas it was mostly negative in those without EM. Cytotoxic T cell (CD8+CD11b-) profiles in PF with EM were also quiet different from those without EM. Cellular immunity in the peripheral blood did not change during the course of IVF-ET cycles, although plasma levels of ovarian steroid hormones significantly increased comparing with that in normal ovarian cycles. Cytotoxic T cell type 1 (Tc1) profiles might be useful predictive values in the pregnancy outcome for infertile patients with EM. PMID- 12770752 TI - Hypermethylation can selectively silence multiple promoters of steroid receptors in cancers. AB - Multiple promoters and differential splicing of 5' upstream exons are often found in various nuclear receptor genes including steroid receptors. Three promoters control the expression of human estrogen receptor alpha (ERalpha) isoforms: ERalpha-A, ERalpha-B, and ERalpha-C, and two promoters control the expression of human progesterone receptor (PR) isoforms: PR-A and PR-B. The expression levels of these isoforms differ with respect to each other in certain target tissues. The role of these isoforms may differ in various types of cells and tissues. The ER and PR contain CpG islands in the 5' upstream regions. One possible mechanism for changing the transcriptional status is methylation of CpG-enriched regions in these isoforms. We have investigated the expression and methylation status of the three different ERalpha promoters and the two different PR gene promoters by using methylation specific PCR (MSP) and direct DNA sequencing in several endometrial and prostate cancer cell lines and tissues. The results of these experiments suggest that ERalpha-A, ERalpha-B, and PR-A were expressed and ERalpha-C and PR-B were inactivated in endometrial cancers. To the contrary, ERalpha-A and ERalpha-B were inactivated and ERalpha-C, PR-A and PR-B were expressed in all prostate cancer. Treatment with demethylating agent (5-aza-2' deoxycytidine) restored these gene expressions, suggesting that inactivation of this gene is through methylation. Our MSP and direct DNA sequencing showed that ERalpha-A, ERalpha-B, and PR-A genes were unmethylated and ERalpha-C and PR-B were methylated in endometrial cancers although ERalpha-A and ERalpha-B were methylated and ERalpha-C, PRA and PRB were unmethylated in prostate cancers. These reports clearly demonstrate that selective hypermethylation can selectively silence multiple promoters of steroid receptors in carcinogenesis. PMID- 12770754 TI - Views on the autoimmunity hypothesis for Chagas disease pathogenesis. AB - Initially, the notion that the pathogenesis of Chagas disease has an autoimmune component was based on the finding that sera from Trypanosoma cruzi-infected patients or laboratory animals contain antibodies that recognize both parasite and host tissue antigens. Subsequent work suggested that T lymphocytes from chagasic patients and animals also displayed such cross-reactivity. However, the autoimmunity hypothesis has remained controversial because of experimental pitfalls, incomplete or inadequate controls, difficulties in reproducing some key results, and a lack of persuasive evidence that the cross-reactive antibodies or lymphocytes can truly effect the multifaceted pathological features of Chagas disease. Whether the immunologic autoreactivities described to date cause chagasic pathology or result from it is another unresolved question. Discussed herein are the most recent contributions to this topic and the reservations they have raised. PMID- 12770755 TI - A bioinformatic approach to the identification of bacterial proteins interacting with Toll-interleukin 1 receptor-resistance (TIR) homology domains. AB - Members of the Toll-like receptor (TLR) family are currently under intense scrutiny for their role in the sampling and recognition of pathogens. It has already been reported that both vaccinia virus and Yersinia spp. express proteins that help them evade the TLR mediated immune response, acting through the Toll interleukin-1 receptor-resistance (TIR) domain and leucine-rich repeat region of the host TLRs respectively. The TIR domain is involved in the dimerisation of the TLRs and their complexation with their adapter molecules. We tested here the hypothesis that bacteria have the ability to secrete proteins containing similar motifs to the intracellular TIR domains that are involved in the TIR-TIR interaction necessary for the subsequent signal transmission. Based upon their sequence homology, proteins expressing TIRs have been divided into three sub classes, based around the TLRs, the TLR adapter proteins, and the interleukin-1 and -18 adapter proteins. The highly conserved regions from these separate sub families were then used to identify similar bacterial proteins. The bacterial proteins identified were then included in an iterative MEME-BLAST process to broaden the search. Tollip, a known TLR antagonist and adapter protein, was included in this investigation although it does not fit into any of the three sub classes outlined above. If suitable bacterial proteins had been identified, it would signify that certain bacteria had evolved a mechanism to aid them in avoiding detection by the innate immune system acting through the TIR domains. At this stage one has to conclude that there is no evidence currently available suggesting such a mechanism, when using the strategy applied here. PMID- 12770756 TI - Trypanosoma cruzi ubiquitin as an antigen in the differential diagnosis of Chagas disease and leishmaniasis. AB - In the present report we describe Trypanosoma cruzi ubiquitin as an antigen to be utilized in the differential diagnosis of Chagas disease and leishmaniasis. Initially, recombinant T. cruzi ubiquitin was evaluated against a panel of sera by phage dot immunoassay, showing a good performance against chagasic sera. However, the presence of a carboxy-terminal tail region encoding a ribosomal protein homologous to a related protein present in the genome of Leishmania sp. gave significant cross-reactivity with leishmanial sera. Therefore, ubiquitin was purified by a simple biochemical protocol and its immunoreactivity was studied by enzyme-linked immunosorbent assay. Analysis of 104 sera indicates that the response to ubiquitin is very sensitive towards chronic chagasic sera (98%) and, more important, highly species-specific, presenting better performance compared to the use of the recombinant protein or the total epimastigote extracts when tested against a panel of leishmanial sera, where out of a total of 70 sera tested, only five sera from the mucocutaneous form of the disease reacted with T. cruzi ubiquitin. On the other hand, Leishmania ubiquitin was not recognized by chagasic sera, but was recognized by sera from different forms of leishmaniasis. These results make ubiquitin an excellent candidate to be used in the differential diagnosis of these two parasitic diseases. The molecular basis for this highly species-specific response is discussed. PMID- 12770757 TI - A murine model of Madurella mycetomatis eumycetoma. AB - Eumycetoma due to Madurella mycetomatis is a major mycological health problem in endemic areas. We infected BALB/c mice (male or female) with various amounts of M. mycetomatis mycelium, containing sterilized soil as a natural adjuvant or Freund's incomplete adjuvant. Mice differed with respect to age and immune status. Intraperitoneal, subcutaneous and intravenous inoculation was explored and survival was monitored. Mice were killed at various intervals after inoculation, checked for the presence of the characteristic black grains, and organs were cultured for M. mycetomatis. Infected organs were subjected to histopathological examination. Immunocompetent male mice were as susceptible as immunocompromised female mice, but showed higher mortality rates. In conclusion, a reproducible mouse model of intraperitoneal M. mycetomatis infection with characteristic black grains in immunocompetent adult or young female mice was developed. Although this experimental model does not simulate macroscopic features of the subcutaneous M. mycetomatis infection in humans, the histopathological characteristics of the lesions and the development of black grains are clearly representative for the human infection. This model will enable further studies on the pathogenesis as well as prevention and treatment of the fungal infection. PMID- 12770758 TI - Immunogenicity and biophysical properties of a Nocardia brasiliensis protease involved in pathogenesis of mycetoma. AB - We isolated and purified to homogeneity a caseinolytic protease from a Nocardia brasiliensis cell extract. Preparative polyacrylamide gel electrophoresis and electroelution were employed for purification. This purified protease was injected in BALB/c mice and induced IgM and IgG anti-protease antibodies. Active immunization of mice with this protease prevented mycetoma development in experimentally infected animals. Passive immunization with hyperimmune sera containing a high anti-protease antibody titer conferred partial but transient protection when collected 30 days after donor's immunization. The protective effect of hyperimmune sera was lost when obtained from donors after 60 days from their immunization despite its higher anti-protease antibody concentration. Cytokines are good candidates to explain these findings. PMID- 12770759 TI - Apotransferrin administration prevents growth of Staphylococcus epidermidis in serum of stem cell transplant patients by binding of free iron. AB - We investigated the effect of free, non-transferrin-bound iron occurring in haematological stem cell transplant patients on growth of Staphylococcus epidermidis in serum in vitro, and prevention of bacterial growth by exogenous apotransferrin. S. epidermidis did not grow in normal serum at inoculated bacterial densities up to 10(3) cfu ml(-1) but slow growth could be detected at higher initial inocula. Addition of free iron abolished the growth-inhibitory effect of serum, whereas addition of apotransferrin again restored it. Appearance of free iron and loss of growth inhibition coincided in patient serum samples taken daily during myeloablative therapy. Intravenously administered apotransferrin effectively bound free iron and restored the growth inhibition in patient sera. The results suggest that exogenous apotransferrin might protect stem cell transplant patients against infections by S. epidermidis and possibly other opportunistic pathogens. PMID- 12770760 TI - Protective efficacy of PspA (pneumococcal surface protein A)-based DNA vaccines: contribution of both humoral and cellular immune responses. AB - Streptococcus pneumoniae is a major public health problem and new strategies for the development of cost-effective alternative vaccines are important. The use of protein antigens such as PspA (pneumococcal surface protein A) is a promising approach to increase coverage at reduced costs. We have previously described the induction of a strong antibody response by a DNA vaccine expressing a C-terminal fragment of PspA. Fusion of this fragment with the cytoplasmic variant of SV40 large T-antigen (CT-Ag) caused reduction in specific interferon-gamma produced by stimulated spleen cells. In this work we show that the DNA vaccine expressing the C-terminal region of PspA elicits significant protection in mice against intraperitoneal challenge with a virulent strain of S. pneumoniae. Furthermore, fusion with CT-Ag completely abrogated the protection elicited by DNA immunization with this fragment. In this case, protection did not correlate with total anti-PspA antibody production nor with total IgG2a levels. The anti-PspA sera obtained from both constructs showed equivalent opsonic activity of pneumococci, indicating that the antibodies produced were functional. We could, though, observe a correlation between a lower IgG1:IgG2a ratio, which is indicative of a stronger bias towards Th1 responses, and protection. We also show that a vector expressing the most variable N-terminal alpha-helical region induces higher antibody formation, with increased protection of mice against intraperitoneal challenge with a more virulent strain of S. pneumoniae. As a whole, these results indicate that antibodies elicited against PspA would not be solely responsible for the protection induced by DNA vaccination and that cell mediated immune responses could also be involved in protection against pneumococcal sepsis. PMID- 12770762 TI - Infectious exacerbations of chronic obstructive pulmonary disease associated with respiratory viruses and non-typeable Haemophilus influenzae. AB - Infectious exacerbations of chronic obstructive pulmonary disease (COPD) have been reported to occur with both viral and bacterial pathogens. In this study, 35 exacerbations associated with the isolation of non-typeable Haemophilus influenzae from sputum were identified as part of a prospective longitudinal study. Samples from these patients were subjected to immunoassays to identify a new immune response to the homologous isolate of non-typeable H. influenzae to more accurately assess a bacterial etiology. These patients also were studied carefully for evidence of viral infection using viral culture, serology and polymerase chain reaction-based assays. Sixteen of 35 exacerbations (45.7%) were associated with evidence of acute viral infection and 11 of the 35 exacerbations (31.4%) were associated with the development of new serum IgG to homologous non typeable H. influenzae. Overall, evidence of infection with a respiratory virus or non-typeable H. influenzae was seen in 24 of 35 exacerbations (68.6%). No association between viral infection and immune response to non-typeable H. influenzae was observed, although a trend toward an immune response to non typeable H. influenzae and absence of viral infection was seen. The results show that exacerbations in adults with COPD were associated with infection caused by virus alone, non-typeable H. influenzae alone, or virus and non-typeable H. influenzae simultaneously. PMID- 12770761 TI - Serological characterization of anti-endotoxin serum directed against the conjugate of oligosaccharide core of Escherichia coli type R4 with tetanus toxoid. AB - The covalent conjugate of oligosaccharide core of Escherichia coli type R4 with tetanus toxoid was prepared using reaction of reductive amination. The neoglycoconjugate was a good immunogen in rabbits yielding a high level of anti lipopolysaccharide (LPS) antibodies of the IgG class. It was found that antiserum was able to react with the smooth LPS molecules of identical (R4) or related (R1) core type. The reactions were shown in the enzyme-linked immunosorbent assay and the immunoblotting test. Flow cytometry showed that anti-core antibodies reacted with LPS present on intact, live, smooth bacteria labelling more than 90% of cells. The anti-OS R4-TT serum used for in vitro studies showed high endotoxin neutralization activity. The serum inhibited endotoxin-induced tumor necrosis factor alpha and nitric oxide synthesis by the J-774A.1 cell line and attenuated pulmonary retention of YAC-1 cells. PMID- 12770763 TI - HIV-1 and its transmembrane protein gp41 bind to different Candida species modulating adhesion. AB - Oral candidiasis in HIV-1-infected individuals is widely believed to be triggered by the acquired T-lymphocyte immunodeficiency. Recently, binding of the HIV-1 envelope protein gp160 and its subunit gp41, and also of the whole virus itself, to Candida albicans has been shown. The present study shows that, in addition to C. albicans, HIV-1 gp41 also binds to yeast and hyphal forms of Candida dubliniensis, a species which is closely related to C. albicans, and to Candida tropicalis but not to Candida krusei, Candida glabrata or Saccharomyces cerevisiae. The previous finding that gp41 binding to C. albicans augments fungal virulence in vitro is supported by the observation that the yeast showed an enhanced adhesion to HIV-infected H9 cells in comparison to uninfected cells. In line with these results soluble gp41 itself reduced binding of C. albicans to both endothelial and epithelial cell lines, confirming a dominant role of the gp41 binding moiety on the surface of Candida for adhesion. Surface-associated secreted aspartic proteinases (Saps) play an important role in candidial adhesion, but are not likely to be involved in the interaction as gp41 binding to the C. albicans parental wild-type strain was comparable to that of three different isogenic Sap deletion mutants. Furthermore, gp41 binding to the yeast killer toxin-susceptible C. albicans strain 10S was not inhibitable by an anti YKT receptor antibody. In conclusion, HIV-1 interacts with different clinically important Candida spp., and may thereby affect the outcome of the respective fungal infection. PMID- 12770764 TI - The nucleotide-sugar transporter family: a phylogenetic approach. AB - Nucleotide sugar transporters (NST) establish the functional link of membrane transport between the nucleotide sugars synthesized in the cytoplasm and nucleus, and the glycosylation processes that take place in the endoplasmic reticulum (ER) and Golgi apparatus. The aim of the present work was to perform a phylogenetic analysis of 87 bank annotated protein sequences comprising all the NST so far characterized and their homologues retrieved by BLAST searches, as well as the closely related triose-phosphate translocator (TPT) plant family. NST were classified in three comprehensive families by linking them to the available experimental data. This enabled us to point out both the possible ER subcellular targeting of these transporters mediated by the dy-lysine motif and the substrate recognition mechanisms specific to each family as well as an important acceptor site motif, establishing the role of evolution in the functional properties of each NST family. PMID- 12770765 TI - Galactosyltransferase--still up and running. AB - The following review on galactosyltransferase (gal-T1) intends to cover genetic, biochemical, structural, biotechnological, cell biological and medical aspects of this enzyme in a comprehensive manner from discovery to the present day which have brought to light a genetic defect of this enzyme. Early work has only been included if it appeared relevant to ongoing issues. Following the evolution of a research topic over 40 years is in itself a fascinating endeavor as it permits to observe the ins and outs of hypotheses, fashions and errors. Gal-T1 is a beautiful example as it has been involved in almost every aspect of life science. Importantly, there is a future to this enzyme as a research topic, since many questions still remain unanswered: to which extent is it a representative Golgi protein? What is the role of the gene family of gal-Ts? Does gal-T1 exert any functions other than a catalytic one? Why is it phosphorylated? Does it form homodimers in vivo? Surely, there is room for further work, which is likely to reveal further insights into cellular trafficking and signaling and, in the context of the gene family, shall contribute to understanding development and morphogenesis. PMID- 12770766 TI - Enzymatic synthesis of the core-2 sialyl Lewis X O-glycan on the tumor-associated MUC1a' peptide. AB - Starting from a tumor-associated synthetic MUC1-derived peptide MUC1a' and using a completely enzymatic approach for the synthesis of the core-2 sialyl Lewis X glycopart, the following glycopeptide was synthesized: AHGV[Neu5Ac(alpha2 3)Gal(beta1-4)[Fuc(alpha1-3)]GlcNAc(beta1-6)[Gal(beta1-3)]GalNAc(alpha1 O)]TSAPDTR. First, polypeptide N-acetylgalactosaminyltransferase 3 was used to site-specifically glycosylate MUC1a' to give MUC1a'-GalNAc. Then, in a one-pot reaction employing beta-galactosidase and core-2 beta6-N acetylglucosaminyltransferase the core-2 O-glycan structure was prepared. The core-2 structure was then sequentially galactosylated, sialylated, and fucosylated by making use of beta4-galactosyltransferase 1, alpha3 sialyltransferase 3, and alpha3-fucosyltransferase 3, respectively, resulting in the sialyl Lewis X glycopeptide. The overall yield of the final compound was 23% (3.2 mg, 1.4 micromol). During the synthesis three intermediate glycopeptides containing O-linked GalNAc, Gal(beta1-4)GlcNAc(beta1-6)[Gal(beta1-3)]GalNAc, and Neu5Ac(alpha2-3)Gal(beta1-4)GlcNAc(beta1-6)[Gal(beta1-3)]GalNAc, respectively, were isolated in mg quantities. All products were characterized by mass spectrometry and NMR spectroscopy. PMID- 12770767 TI - The role of glucosidase II and endomannosidase in glucose trimming of asparagine linked oligosaccharides. AB - This review covers various aspects of glucose trimming reactions occurring on asparagine-linked oligosaccharides. Structural and functional features of two enzymes, glucosidase II and endo-alpha-mannosidase, prominently involved in this process are summarized and their striking differences in terms of substrate specificities are highlighted. Recent results of analyses by immunoelectron microscopy of their distribution pattern are presented which demonstrate that glucose trimming is not restricted to the endoplasmic reticulum (ER) but additionally is a function accommodated by the Golgi apparatus. The mutually exclusive subcellular distribution of glucosidase II and endomannosidase are discussed in terms of their significance for quality control of protein folding and N-glycosylation. PMID- 12770768 TI - Glycosylation of hepatitis C virus envelope proteins. AB - Enveloped viruses are surrounded by a membrane derived from the host-cell that contains proteins called "envelope proteins". These proteins play a major role in virus assembly and entry. In most of the enveloped viruses, they are modified by N-linked glycosylation which is supposed to play a role in their stability, antigenicity and biological functions. Glycosylation is also known to play a major role in the biogenesis of proteins by being directly and/or indirectly involved in protein folding. Recent studies on hepatitis C virus (HCV) envelope proteins have revealed a complex interplay between cleavage by signal peptidase, folding and glycosylation. The knowledge that has been accumulated on the early steps of glycosylation of these proteins is presented in this review. PMID- 12770769 TI - Chemical modifications of alpha1,6-fucosyltransferase define amino acid residues of catalytic importance. AB - alpha1,6-Fucosyltransferase (alpha6FucT) of human platelets was subjected to the action of phenylglyoxal (PLG), pyridoxal-5'-phosphate/NaBH(4) (PLP), and diethyl pyrocarbonate (DEPC) the reagents that selectively modify the structure of amino acids arginine, lysine and histidine, respectively, as well as to N ethylmaleimide (NEM), mersalyl, p-chloromercuribenzoate (pCMB), iodoacetate, iodoacetamide, and methyl iodide that react with sulfhydryl group of cysteine. In addition, we treated the enzyme with beta-mercaptoethanol, a reagent that disrupts disulfide bonds. All reagents except NEM significantly inactivated alpha6FucT. Protection against the action of PLG, PLP and sulfhydryl modifying reagents was offered by GDP-fucose, GDP, and the acceptor substrate, a transferrin-derived biantennary glycopeptide with terminal GlcNAc residues. Neither donor nor acceptor substrate offered, however, any protection against inactivation by DEPC or beta-mercaptoethanol. We conclude that arginine, cysteine and probably lysine residues are present in, or closely by, the donor and acceptor substrate binding domains of the enzyme, whereas histidine may be a part of its catalytic domain. However, the primary structure of alpha6FucT does not show cysteine residues in proximity to the postulated GDP-fucose-binding site and acceptor substrate binding site of the enzyme that contains two neighboring arginine residues and one lysine residue (Glycobiol. 10 (2000) 503). To rationalize our results we postulate that platelet alpha6FucT is folded through disulfide bonds that bring together donor/acceptor-binding- and cysteine- and lysine-rich, presumably acceptor substrate binding sites, thus creating a catalytic center of the enzyme. PMID- 12770770 TI - Recognition of cell surface acceptors by two human alpha-2,6-sialyltransferases produced in CHO cells. AB - The action of sialyltransferases (STs) on cell surface glycoconjugates is a key process in shaping cell phenotype in a variety of cells mostly involved in migratory and adhesive pathways. The factors determining cell-specific pattern of glycosylation are so far poorly understood. Most STs are resident proteins of the Golgi apparatus, where acceptors are sialylated while they are in transit to the cell surface. To identify putative structural features that may account for their acceptor preference, we analyzed 53 cloned animal and human STs. We could identify conserved regions and peptide motifs representative of ST subfamilies, located at the C-terminal end of the hypervariable region upstream from the L sialyl motif. Residues 93-100 in human ST6Gal I (hST6Gal I) were shown to be crucial for enzymatic activity when deleted and expressed in CHO cells. The Delta100 hST6Gal I mutant protein was fully recognized by polyclonal anti-hST6Gal I antibodies and followed the intracellular secretory pathway. This indicated that the conserved QVWxKDS sequence is essential for the whole catalytic domain to acquire a biologically active conformation. When full-length epitope-tagged hST6Gal I and hST6GalNAc I constructs were transfected in CHO cells, the alpha 2,6 sialylated glycotope was found to be largely restricted to intracellular resident acceptors and enzymatic activity based on fluorescent lectin staining. In contrast, both enzymes deprived of their membrane anchor and part of the hypervariable region but still possessing the conserved domains exhibited a very efficient transfer of sialic acid to cell surface glycoconjugates. Colocalization of the ST6Gal I mutant proteins with early and late Golgi markers such as giantin or rab6 proteins confirmed that soluble STs migrate forward in these subcompartments where they can act upon newly synthesized acceptors and follow the secretory pathway. It is thus concluded that downstream from the transmembrane domain, native STs possess peptide sequences that allow them to sialylate glycoprotein acceptors selectively along their transit within Golgi stacks. PMID- 12770771 TI - Involvement of glycosylation in the intracellular trafficking of glycoproteins in polarized epithelial cells. AB - The surface of epithelial cells is composed of apical and basolateral domains with distinct structure and function. This polarity is maintained by specific sorting mechanisms occurring in the Trans-Golgi Network. Peptidic signals are responsible for the trafficking via clathrin-coated vesicles by means of an interaction with an adaptor complex (AP). The basolateral targeting is mediated by AP-1B, which is specifically expressed in epithelial cells. In contrast, the apical targeting is proposed to occur via apical raft carriers. It is thought that apically targeted glycoproteins contain glycan signals that would be responsible for their association with rafts and for apical targeting. However, the difficulty in terms of acting specifically on a single step of glycosylation did not allow one to identify such a specific signal. The complete inhibition of the processing of N-glycans by tunicamycin often results in an intracellular accumulation of unfolded proteins in the Golgi. Similarly, inhibition of O glycosylation can be obtained by competitive substrates which gave a complex pattern of inhibition. Therefore, it is still unknown if glycosylation acts in an indirect manner, i.e. by modifying the folding of the protein, or in a specific manner, such as an association with specific lectins. PMID- 12770772 TI - Regulation of the intestinal glycoprotein glycosylation during postnatal development: role of hormonal and nutritional factors. AB - This review focuses on the regulation of the glycoprotein glycosylation process in small intestine and colon during postnatal development. Glycoproteins play a prominent part in intestine as mucins secreted by the goblet cells and as molecules of biological interest largely present in the microvillus membrane of the enterocytes (digestive enzymes, transporters). The age-related changes in the intestinal glycosylation control the quality of glycan chains of glycoproteins. Postnatal maturation is observed at all stages of the glycoprotein glycosylation. But it is essentially characterised in the external glycosylation by a shift from sialylation to fucosylation depending on the transcriptional regulation of the corresponding glycosyltransferases, but also on coordinate changes in the activities of glycosyltransferases and of their regulatory proteins, in nucleotide-sugar bioavailability and in product degradation by oxidases. Many factors have been evoked to trigger these changes, among which are hormonal (glucocorticoids, insulin) and dietary factors. Changes in the structure of the glycoprotein glycans might be important for the transport, the barrier function, the implantation of the immune defences and of the microflora and even probably for the biological activity of some digestive enzymes. PMID- 12770773 TI - Developmentally regulated biosynthesis of carbohydrate and storage polysaccharide during differentiation and tissue cyst formation in Toxoplasma gondii. AB - Toxoplasma gondii belongs to the Apicomplexa phylum, which comprises protozoan parasites of medical and veterinary significance, responsible for a wide variety of diseases in human and animals, including malaria, toxoplasmosis, coccidiosis and cryptosporidiosis. During infection in the intermediate host, T. gondii undergoes stage conversion between the rapidly replicating tachyzoite that is responsible for acute toxoplasmosis and the dormant or slowly dividing encysted bradyzoite. The tachyzoite-bradyzoite interconversion is central to the pathogenic process and is associated with the life-threatening recrudescence of infection observed in immunocompromised patients such as those suffering from AIDS. In chronic infections, the bradyzoites are located within tissue cysts found predominantly in brain and muscles. The tissue cyst is enclosed by a wall containing specific lectin binding sugars while the bradyzoites have accumulated large amounts of the storage polysaccharide of glucose, amylopectin. Our recent findings have identified several genes and proteins associated with amylopectin synthesis or degradation and glucose metabolism, including different isoforms of certain glycolytic enzymes, which are stage-specifically expressed during tachyzoite-bradyzoite interconversion. Here, we will discuss how the genes and enzymes involved in carbohydrate metabolisms are used as molecular and biochemical tools for the elucidation of molecular mechanisms controlling T. gondii stage interconversion and cyst formation. PMID- 12770774 TI - Alpha1,3fucosyltransferases in cystic fibrosis airway epithelial cells. AB - Cystic fibrosis (CF) has a glycophenotype of aberrant sialylation and/or fucosylation. The CF glycophenotype is expressed on membrane glycoconjugates of CF airway epithelial cells as increased fucosyl residues in alpha1,3/4 linkage to N-acetyl glucosamine, decreased fucosyl residues in alpha1,2 linkage to galactose and decreased sialic acid. To define the cause of this phenotype, the enzyme activity of alpha1,3fucosyltransferase (FucT) was examined in extracts of CF airway epithelial cells with a variety of low molecular weight substrates. Using Galbeta1,4GlcNAc as substrate, the activity was divided into 66% alpha1,3FucT and 34% alpha1,2FucT. mRNA expression examined with probes to FucTIII, IV, and VII showed that the highest expression of two CF cell lines was for FucTIV. Only one CF cell line expressed mRNA for FucTIII. The non CF airway epithelial cells had significant enzyme activity for alpha1,3FucT and strong mRNA expression for FucTIV. Thus as reported previously for alpha1,2FucT, the biochemical capacity for alpha1,3FucT was present in both the CF and non CF cells and can not be the cause of the CF glycophenotype. These results support the hypothesis that wild type CFTR acts in the Golgi and when mutated as in CF, faulty compartmentalization of terminal glycosyltransferases results, yielding the CF glycophenotype. PMID- 12770775 TI - Sulfated oligosaccharides isolated from the respiratory mucins of a secretor patient suffering from chronic bronchitis. AB - The most acidic carbohydrate chains released by alkaline borohydride treatment of the bulk of airway mucins secreted by a patient (blood group O, secretor) suffering from a mildly infected chronic bronchitis have been fractionated using high-performance anion-exchange chromatography (HPAEC) according to a protocol already described [Lo-Guidice et al., J. Biol. Chem. 269 (1994) 18794] and were analyzed using 1H-NMR spectroscopy and matrix-assisted laser-adsorption-time-of flight (MALDI-TOF) spectrometry. Many fractions corresponded to mixtures of oligosaccharides. This confirmed the wide diversity of the post-translational processes involved in the biosynthesis of airway mucins, which had already been observed in bronchial diseases, such as chronic bronchitis and cystic fibrosis (CF). Seven fractions were directly purified by HPAEC, allowing their structural determination. Six of them corresponded to 3-O-sulfated oligosaccharide chains terminated by a sulfated N-acetyllactosamine, a sulfated Lewis X or a sulfated Lewis A determinant, and the last one corresponded to a 6-O-sulfated chain terminated by a sulfated H-2 determinant. Three oligosaccharides had core type 2 and the other four had core type 4: IIIc2-9: Gal(beta1-3)[HSO(3)-3-Gal(beta1 4)GlcNAc(beta1-6)]GalNAc-ol, IIIc2-10: Gal(beta1-3)[Fuc(alpha1-2)Gal(beta1 4)[HSO(3)-6-]GlcNAc(beta1-6)]GalNAc-ol, IIIc2-4: Fuc(alpha1-2)Gal(beta1-3)[HSO(3) 3-Gal(beta1-4)[Fuc(alpha1-3)]GlcNAc(beta1-6)]GalNAc-ol, IIIc2-8: Fuc(alpha1 2)Gal(beta1-3)GlcNAc(beta1-3)[HSO(3)-3-Gal(beta1-4)GlcNAc(beta1-6)]GalNAc-ol, IIIc2-7: Fuc(alpha1-2)Gal(beta1-3)GlcNAc(beta1-3)[Gal(beta1-4)[HSO(3)-6 ]GlcNAc(beta1-6)]GalNAc-ol, IIIc2-3: Fuc(alpha1-2)Gal(beta1-3)GlcNAc(beta1 3)[HSO(3)-3-Gal(beta1-4)[Fuc(alpha1-3)]GlcNAc(beta1-6)]GalNAc-ol, IIIc1-4: Fuc(alpha1-2)Gal(beta1-3)GlcNAc(beta1-3)[HSO(3) -3-Gal(beta1-3)[Fuc(alpha1 4)]GlcNAc(beta1-3)Gal(beta1-4)GlcNAc(beta1-6)]GalNAc-ol. Like previous data concerning the airway mucins from another patient (blood group O and non secretor) suffering from chronic bronchitis [Lo-Guidice et al., Glycoconj. J. 14 (1997) 113], no disialylated oligosaccharide and no sialylated and sulfated oligosaccharide bearing sialyl Lewis X epitope could be isolated. This is in contrast with the data obtained with the airway mucins secreted by the patient severely infected by Pseudomonas aeruginosa and suffering from CF, suggesting that important differences occur in the biosynthesis of airway mucins secreted by patients suffering from different bronchial diseases with or without severe infection. PMID- 12770776 TI - A high-density putative monomeric mucin is the major [35S]labelled macromolecular product of human colorectal mucins in organ culture. AB - We have studied the biosynthesis of mucins in organ cultures of human colon using isopycnic density-gradient centrifugation following pulse labelling with [(35)S]sulphate and [(3)H]-D-glucosamine. A high-density [(35)S]sulphate labelled component, of larger size than MUC2 monomers, appeared in the tissue and also in the medium. It was not degraded by reduction, trypsin digestion, digestion with chondroitin ABC lyase or heparan sulphate III lyase, but was cleaved into smaller fragments following alkaline borohydride treatment and appears to be a monomeric, mucin-like molecule containing a protease-resistant domain with a larger hydrodynamic volume than MUC2 monomers. Although this macromolecule incorporated much more radiolabel than MUC2, it was not detected using chemical analysis and thus appears to be a component with a high metabolic turnover present in a very small amount. Most of the [(3)H]-D-glucosamine label was associated with low density material that was well separated from MUC2, which was poorly labelled. Most of MUC2 was associated with the tissue as an 'insoluble' complex. The amount of MUC2 remained constant and its associated radiolabel increased only slightly with time. Analysis of the MUC2 subunits from the reduced 'insoluble' complex showed the typical reduction-insensitive oligomers and confirmed that the radiolabel was associated with this mucin. The large size of the [(35)S]-labelled putative monomeric mucin makes it difficult to separate it from reduced insoluble complex MUC2. As a result, many studies of intestinal mucin synthesis and secretion in the past have most likely been performed on 'mixtures' of this mucin and MUC2 and are thus not possible to interpret as the metabolic behaviour of oligomeric mucins. PMID- 12770777 TI - Isolation of null alleles of the Caenorhabditis elegans gly-12, gly-13 and gly-14 genes, all of which encode UDP-GlcNAc: alpha-3-D-mannoside beta1,2-N acetylglucosaminyltransferase I activity. AB - UDP-GlcNAc: alpha-3-D-mannoside beta1,2-N-acetylglucosaminyltransferase I (GnT I) is a Golgi-resident enzyme which transfers a GlcNAc residue in beta1,2 linkage to the Manalpha1,3Manbeta-terminus of (Manalpha1,6(Manalpha1,3)Manalpha1,6)(Manalpha1,3)Manbeta1,4GlcNAcbeta1,4GlcNAc Asn-protein, thereby initiating the synthesis of hybrid N-glycans. Three Caenorhabditis elegans genes homologous to mammalian GnT I (designated gly-12, gly-13 and gly-14) have been cloned. All three cDNAs encode proteins with GnT I enzyme activity. We report in this paper the preparation by ultra-violet (UV) light irradiation in the presence of trimethylpsoralen, of mutants lacking either gly-12, gly-13 or gly-14. A double null mutation in the gly-12 and gly-14 genes (gly-14; gly-12) has also been prepared. These mutations are intragene deletions, removing large portions of the GnT I catalytic domain, and are therefore, all molecular nulls. The gly-12 and gly-14 mutants as well as the gly-14; gly-12 double mutant all displayed wild-type phenotypes, indicating that neither gly-12 nor gly-14 is necessary for worm development under standard laboratory conditions. In contrast, about 60% of the mutants lacking the gly-13 gene arrested as L1 larvae at 20 degrees C and the remaining 40% homozygous worms grew to adulthood but displayed severe morphological and behavioral defects despite the presence of the other two GnT I genes, gly-12 and gly-14. Attempts to rescue the gly-13 null phenotype with the wild type transgene were not successful. However, lethality co-segregated with the gly-13 deletion within 0.02 map units (mu) in genetic mapping experiments, suggesting that the gly-13 mutation is responsible for the phenotype. PMID- 12770778 TI - Determination of the half-life of the murine beta4-galactosyltransferase-1 mRNA in somatic cells using the tetracycline-controlled transcriptional regulation system. AB - The glycosyltransferases are recognized as a functional family of an estimated 300 distinct, intracellular, membrane-bound enzymes that are positioned along the secretory pathway and participate coordinately in the biosynthesis of the carbohydrate moieties on glycoconjugates. The full-length cDNA sequence for many of these proteins is now available yet little is known about the transcriptional or translational regulation of a given transcript or its decay rate in the cell. These issues are made more complex by the observations that transcription of a glycosyltransferase gene in different cells/tissues results in mRNAs with significantly different structures. For example, transcription of the murine beta4-galactosyoltransferase-1 gene in somatic cells yields two transcripts of 3.9 and 4.1 kb. In contrast transcription of this gene in developing male germ cells results in transcripts of 2.9 and 3.1 kb which are distinguished from their somatic cell counterparts primarily by the deletion of approximately 1.7 kb of sequence in the 3'-untranslated region (UTR). With the long range goal of determining the role that the 3'-UTR serves in mRNA decay we have taken advantage of a recently developed methodology, the Tet-Off system, to determine the half life of the mRNA encoding beta4-galactosyltransferase-1 in the murine NIH 3T3 somatic cell line. We show that the beta4-galactosyltransferase-1 mRNA has a half life of approximately 84 min (range of 82-85 min) in 3T3 cells and that substitution of the galactosyltransferase coding sequence with the coding sequence of luciferase does not significantly alter the decay rate (approximately 87 min; range of 84-91 min). This latter observation suggests that the beta4 galactosyltransferase-1 coding sequence does not contain functional elements that affect the intrinsic stability of this mRNA. PMID- 12770779 TI - A set of glycoproteins recognized by A2B5 antibody with major bands at 55 and 76 kDa is overexpressed in human head and neck squamous cell carcinomas. AB - A2B5 antibody was found to strongly label frozen sections of human head and neck squamous cell carcinomas. The low amount of glycolipids (c-series gangliosides and sulfatides) purified from the same tumors and reactive with A2B5 by immunostaining on thin-layer plates could not account for the high level of tissue labeling. Proteins were extracted from both normal tissues and squamous cell carcinomas and analyzed by Western blot with A2B5 antibody on PVDF membranes. The antibody was found to stain a set of glycoproteins with two major bands at 55 and 76 kDa present in normal tissues and overexpressed in carcinomas. Staining was abolished by prior treatment of the PVDF membranes either with Arthrobacter ureafaciens neuraminidase or with a solution of 10 mM periodate that is known to destroy carbohydrates. Our results show that the carbohydrate epitope recognized by A2B5 antibody can be displayed by both glycolipids and glycoproteins. PMID- 12770781 TI - Salvage pathways in glycosphingolipid metabolism. AB - In this review, the focus is on the role of salvage pathways in glycosphingolipid, particularly, ganglioside metabolism. Ganglioside de novo biosynthesis, that begins with the formation of ceramide and continues with the sequential glycosylation steps producing the oligosaccharide moieties, is briefly outlined in its enzymological and cell-topological aspects. Neo-synthesized gangliosides are delivered to the plasma membrane, where their oligosaccharide chains protrude toward the cell exterior. The metabolic fate of gangliosides after internalization via endocytosis is then described, illustrating: (a) the direct recycling of gangliosides to the plasma membrane through vesicles gemmated from sorting endosomes; (b) the sorting through endosomal vesicles to the Golgi apparatus where additional glycosylations may take place; and (c) the channelling to the endosomal/lysosomal system, where complete degradation occurs with formation of the individual sugar (glucose, galactose, hexosamine, sialic acid) and lipid (ceramide, sphingosine, fatty acid) components of gangliosides. The in vivo and in vitro evidence concerning the metabolic recycling of these components is examined in detail. The notion arises that these salvage pathways, leading to the formation of gangliosides and other glycosphingolipids, sphingomyelin, glycoproteins and glycosaminoglycans, represent an important saving of energy in the cell economy and constitute a relevant event in overall ganglioside (or glycosphingolipid, in general) turnover, covering from 50% to 90% of it, depending on the cell line and stage of cell life. Sialic acid is the moiety most actively recycled for metabolic purposes, followed by sphingosine, hexosamine, galactose and fatty acid. Finally, the importance of salvage processes in controlling the active concentrations of ceramide and sphingosine, known to carry peculiar bioregulatory/signalling properties, is discussed. PMID- 12770780 TI - Expression of eukaryotic glycosyltransferases in the yeast Pichia pastoris. AB - The methylotrophic yeast Pichia pastoris is often used as an organism for the heterologous expression of proteins and has been used already for production of a number of glycosyltransferases involved in the biosynthesis of N- and O-linked oligosaccharides. In our recent studies, we have examined the expression in P. pastoris of Arabidopsis thaliana and Drosophila melanogaster core alpha1,3 fucosyltransferases (EC 2.4.1.214), A. thaliana beta1,2-xylosyltransferase (EC 2.4.2.38), bovine beta1,4-galactosyltransferase I (EC 2.4.1.38), D. melanogaster peptide O-xylosyltransferase (EC 2.4.2.26), D. melanogaster and Caenorhabditis elegans beta1,4-galactosyltransferase VII (SQV-3; EC 2.4.1.133) and tomato Lewis type alpha1,4-fucosyltransferase (EC 2.4.1.65). Temperature, cell density and medium formulation have varying effects on the amount of activity resulting from expression under the control of either the constitutive glyceraldehyde-3 phosphate dehydrogenase (GAP) or inducible alcohol oxidase (AOX1) promoters. In the case of the A. thaliana xylosyltransferase these effects were most pronounced, since constitutive expression at 16 degrees C resulted in 30-times more activity than inducible expression at 30 degrees C. Also, the exact nature of the constructs had an effect; whereas soluble forms of the A. thaliana xylosyltransferase and fucosyltransferase were active with N-terminal pentahistidine tags (in the former case facilitating purification of the recombinant protein to homogeneity), a C-terminally tagged form of the A. thaliana fucosyltransferase was inactive. In the case of D. melanogaster beta1,4 galactosyltransferase VII, expression with a yeast secretion signal yielded no detectable activity; however, when a full-length form of the enzyme was introduced into P. pastoris, an active secreted form of the protein was produced. PMID- 12770782 TI - Physiological relevance of sphingolipid activator proteins in cultured human fibroblasts. AB - The physiological degradation of several membrane-bound glycosphingolipids (GSLs) by water-soluble lysosomal exohydrolases requires the assistance of sphingolipid activator proteins (SAPs). Four of these SAPs are synthesized from a single precursor protein (prosaposin). Inherited deficiency of this precursor results in a rare disease in humans with an accumulation of ceramide (Cer) and glycolipids such as glucosylceramide and lactosylceramide (LacCer). In a previous study, we have shown that human SAP-D stimulates the lysosomal degradation of Cer in precursor deficient cells. In order to study the role of SAPs (or saposins) A-D in cellular GSL catabolism, we recently investigated the catabolism of exogenously added [(3)H]labeled ganglioside GM1, Forssman lipid, and endogenously [(14)C]labeled GSLs in SAP-precursor deficient human fibroblasts after the addition of recombinant SAP-A, -B, -C and -D. We found that activator protein deficient cells are still able to slowly degrade gangliosides GM1 and GM3, Forssman lipid and globotriaosylceramide to a significant extent, while LacCer catabolism critically depends on the presence of SAPs. The addition of either of the SAPs, SAP-A, SAP-B or SAP-C, resulted in an efficient hydrolysis of LacCer. PMID- 12770783 TI - Early variations of the disialoganglioside GD3 in chicken embryonic brain support its role in cell migration. AB - In the present study a primary culture system of chicken embryo brain neurons was used in the early period of chicken brain development from day 6 until day 8, which was shown to be a suitable model of neuritogenesis, cell migration and reaggregation. Dissociated chicken optic tectum cells from embryonic stage 31 were cultured on polylysine-coated dishes under serum-free conditions up to 3 days. Freshly dissociated neurons developed short processes, which contacted one another and formed fasciculated bundles. Cell somata migrated along the neurite bundles, similar to migrating neurons in vivo, forming three-dimensional tissue like clusters. This system was used to study the possible functions of the disialoganglioside GD3 for these neuronal differentiation steps. GD3 represents the predominant ganglioside of embryonic neurons before neuritogenesis in vitro and in vivo. Its biosynthesis is followed during day 6 until day 8 of embryonic brain development. Incubation of dissociated neurons with the monoclonal antibody R-24, recognising the GD3 on the cell surface, led to a total blocking of neurite outgrowth. Accordingly, neither cell migration nor reaggregation could be found. These results indicate that the disialoganglioside GD3 plays a central role in neuronal differentiation and development in the embryonic chicken brain. PMID- 12770784 TI - Role of tumor-associated gangliosides in cancer progression. AB - Neuroectodermic tumors can mostly be characterized by the presence of tumor associated glycosphingolipid antigens, such as gangliosides, defined by monoclonal antibodies. Recently, cumulative evidence indicates that gangliosides modify the biological effects of several trophic factors, in vitro and in vivo, as well as the mitogenic signaling cascade that these factors generate. The functional roles of gangliosides in tumor progression can be revisited: (i) ganglioside antigens on the cell surface, or shed from the cells, act as immunosuppressors, as typically observed for the suppression of cytotoxic T cells and dendritic cells, (ii) certain gangliosides, such as GD3 or GM2, promote tumor associated angiogenesis, (iii) gangliosides strongly regulate cell adhesion/motility and thus initiate tumor metastasis, (iv) ganglioside antigens are directly connected with transducer molecules in microdomains to initiate adhesion coupled with signaling, and (v) ganglioside antigens and their catabolites are modulators of signal transduction through interaction with tyrosine kinases associated with growth factor receptors or other protein kinases. Given the potential importance of these sialylated gangliosides and their modulating biological behavior in vivo, further studies on the role of gangliosides are warranted. PMID- 12770785 TI - Inhibitors of glycosyl-phosphatidylinositol anchor biosynthesis. AB - Glycosyl-phosphatidylinositol (GPI) is a complex glycolipid structure that acts as a membrane anchor for many cell-surface proteins of eukaryotes. GPI-anchored proteins are particularly abundant in protozoa such as Trypanosoma brucei, Leishmania major, Plasmodium falciparum and Toxoplasma gondii, and represent the major carbohydrate modification of many cell-surface parasite proteins. Although the GPI core glycan is conserved in all organisms, many differences in additional modifications to GPI structures and biosynthetic pathways have been reported. Therefore, the characteristics of GPI biosynthesis are currently being explored for the development of parasite-specific inhibitors. In vitro and in vivo studies using sugars and substrate analogues as well as natural compounds have shown that it is possible to interfere with GPI biosynthesis at different steps in a species specific manner. Here we review the recent and promising progress in the field of GPI inhibition. PMID- 12770787 TI - The induction of heat shock protein 70 in peripheral mononuclear blood cells in elderly patients: a role for inflammatory markers. AB - The induction of heat shock proteins (Hsp) is the response to a plethora of stress signals including hyperthermia, physical stress, and various disease states. Although changes in Hsp expression are associated with certain diseases, the question as to whether this is an adaptation to a particular pathophysiologic state or a reflection of the suboptimal cellular environment associated with the disease remains open. In this study we have investigated the effects of inflammatory mediators on the induction of Hsp 70 in human peripheral mononuclear blood cells using flow cytometry. We demonstrate that without heat shock, the levels of the inflammatory mediators are positively related to Hsp 70 production in monocytes. On the contrary, negative correlations were found between heat induced Hsp 70 production and interleukin-6 (IL-6), as well as various markers of inflammation. These observations are in agreement with the antagonistic effects between heat stress and the inflammatory mediators on the activation of Hsp promoter. PMID- 12770786 TI - Glycosylphosphatidyl-inositols in murine malaria: Plasmodium yoelii yoelii. AB - Glycosylphosphatidyl-inositols (GPIs) are vital major glycoconjugates in intraerythrocytic stages of Plasmodium. Here, we report on the biosynthesis and the characterization of GPIs synthesized by the murine malarial parasite P. yoelii yoelii YM. Parasitized erythrocytes were labeled in vivo and in vitro with either radioactive nucleotide sugar precursors, ethanolamine or glucosamine. The pathway leading to the formation of GPI precursors was found to resemble that described for P. falciparum; however, in P. yoelii, the formation of an additional hydrophilic precursor containing an acid-labile modification was detected. The data suggest that this modification is linked to the fourth mannose attached to the trimannosyl backbone in an alpha1-2 linkage. The modification was susceptible to hydrofluoric acid (HF), but not to nitrous acid (HNO(2)). Data obtained from size-exclusion chromatography on Bio-Gel P4, and Mono Q analysis of the fragments generated by HNO(2) deamination suggest that the modification is due to the presence of an additional ethanolamine linked to the fourth mannose via a phosphodiester bond. PMID- 12770788 TI - V alpha 24+ natural killer T cells are markedly decreased in atopic dermatitis patients. AB - Atopic dermatitis (AD) is a common inflammatory skin disease. In AD, cytokines such as interleukin (IL)-4 or interferon (IFN)-gamma are considered to affect the disease status. Recently, human V alpha 24(+) natural killer T (NKT) cells have been found to produce large amounts of IL-4 and IFN-gamma. Thus there is a possibility that the proportion of V alpha 24(+) NKT cells modifies the AD status. In this study, we examine the proportion of the V alpha 24(+)/V beta 11(+) cells that composes the V alpha 24(+) NKT cells in peripheral blood mononuclear cells (PBMCs) from 71 healthy donors (HDs) and 31 AD patients. Because CD4(-) and CD4(+) NKT subsets show different cytokine production patterns concerning IL-4, these two subsets are evaluated. Our results have shown that the proportion of the V alpha 24(+) NKT cells is markedly reduced in AD patients. In addition, the CD4(-) V alpha 24(+) NKT subset has a tendency to be more reduced than the CD4(+) V alpha 24(+) NKT subset. Moreover, the proportion of CD4(-) V alpha 24(+) NKT(+) cells and Th2 deviation of Th1/Th2 balance is inversely correlated. These observations may contribute to the understanding of the mechanism involved in AD. PMID- 12770789 TI - The detection and definition of IgM alloantibodies in the presence of IgM autoantibodies using flowPRA beads. AB - We have developed a flow cytometry-based screening method using FlowPRA (One Lambda) human leukocyte antigen (HLA) class I panel beads and FlowPRA (One Lambda) HLA class I specificity beads for the detection and definition of immunoglobulin (Ig)M HLA-specific antibodies in the presence of IgM autoantibodies. Forty-six autoantibody-positive patients who were on the waiting list for a renal transplant (56 sera) were tested in parallel with FlowPRA (One Lambda) HLA class I beads and FlowPRA (One Lambda) control beads. Sera that were positive for IgM HLA class I antibodies were subsequently tested with FlowPRA HLA class I specificity beads to determine the HLA specificities. Thirteen of the 46 patients were positive for IgM HLA class I-specific antibodies. Eleven of the 13 had previous failed transplants and 2 were awaiting a primary transplant. For 9 of the 13 positive patients, IgM HLA class I specificities were defined. We have demonstrated the presence of IgM HLA-specific antibodies in patients with IgM autoantibodies. This study demonstrates the value of FlowPRA HLA class I panel and specificity beads for the detection and definition of IgM HLA class I specific antibodies. PMID- 12770790 TI - Differential immunogenicity of paternal HLA Class I antigens in pregnant women. AB - More insight into the differential immunogenicity of human leukocyte antigen (HLA) mismatches will be beneficial for donor selection in clinical transplantation. In this study the immunogenicity of HLA antigens was analyzed by examining the antibody profiles in women who have been pregnant. In total 888 women, who had pregnancy induced HLA alloantibodies, were included in this study, while 413 women who had not been immunized by their pregnancy, served as controls. First it was analyzed whether women expressing particular HLA antigens are more likely to produce HLA alloantibodies. Next we determined whether certain HLA mismatches of their children are more immunogenic than other ones. Finally we studied whether the immunogenicity of specific HLA mismatches is dependent on the HLA phenotype of the women. Women expressing HLA-A3, HLA-A32, and HLA-B21 are more likely to produce alloantibodies whereas women expressing HLA-B13 and HLA B17 have a significantly lower incidence of alloantibodies compared with women expressing other HLA antigens. Children with HLA-A2 or HLA-B5 mismatches induced alloantibodies significantly more often whereas children with HLA-A30, -A31 or A33 and HLA-A28 induced alloantibodies significantly less often than children with other HLA class I mismatches. Finally we could demonstrate that the immunogenicity of a particular HLA mismatch is dependent on the HLA phenotype of the women. Information on the differential immunogenicity of HLA mismatches may be of benefit for the determination of acceptable and taboo mismatches in the case of donor selection for (highly sensitized) patients. PMID- 12770791 TI - Diversity within the DRB1*08 allele family in four populations from a United States hematopoietic stem cell donor database and characterization of five novel DRB1*08 alleles. AB - The frequencies of DRB1*08 alleles within four major United States populations found within a hematopoietic stem cell volunteer donor database were determined by DNA sequencing of over 60 DRB1*08 positive individuals from each group. Seven of 30 known DRB1*08 alleles were identified within this study population (080101, 080201, 080302, 080401, 0806, 0807, and 0811). Each ethnic group was characterized by a different highly prevalent allele: DRB1*080101 in Caucasians; DRB1*080401 in African-Americans; DRB1*080302 in Asians; and DRB1*080201 in Hispanics. The alleles DRB1*080101, DRB1*080201, and DRB1*080401 were present in all four populations. This report also describes five novel DRB1*08 alleles uncovered during routine human leukocyte antigen typing. PMID- 12770792 TI - TNF and TNF receptor polymorphisms in Korean Behcet's disease patients. AB - Behcet's disease (BD) is an autoimmune disease characterized by recurrent oral ulcers, genital ulcers, erythema nodosum, and uveitis. Genetic factors are considered important in its pathogenesis. The serum level of tumor necrosis factor (TNF) is elevated in patients with active BD, and its production is elevated in monocytes and in the gamma delta T cells of BD patients. A dramatic response to anti-TNF-alpha antibody treatment further supports the role of TNF in BD. In this study, we investigated genetic polymorphisms of TNF alpha -308 G/A, TNF beta +252 G/A, and TNFR2 196 R/M in 94 Korean BD patients and age- and sex matched healthy controls to investigate the role of TNF and TNF receptor polymorphisms in BD. The polymerase chain reaction-restriction fragment length polymorphism was used to identify the TNF-alpha promoter (G = TNFA1, A = TNFA2) and TNF-beta intron polymorphisms (G = TNFB1, A = TNFB2), and polymerase chain reaction-singly-strand conformation polymorphism was used to identify TNFR2 196R/M polymorphism (T = TNFR2M, G = TNFR2R). No differences were found in the TNF-alpha, TNF-beta or TNFR2 polymorphisms of the patients and the healthy controls. The allele frequencies of TNFA1/A2 were 0.94/0.06 in patients and 0.96/0.04 in healthy controls (p = 0.36, OR = 0.65, 95% CI = 0.26-1.63), for TNFB1/TNFB2 these were 0.42/0.58 in patients and 0.44/0.56 in controls (p = 0.68, OR = 0.91, 95% CI = 0.61-1.38), and for TNFR2R/TNFR2M 0.23/0.77 in patients and 0.21/0.79 in controls (p = 0.62, OR = 1.13, 95% CI = 0.69-1.84). In conclusion, this study found no differences of TNF alpha -308 G/A, TNF beta +252 G/A or of the TNFR2 196R/M polymorphisms in Korean BD patients versus healthy controls. These findings suggest that the role of TNF in BD is not genetically determined, but can be functionally explained. PMID- 12770793 TI - Mannose binding lectin gene polymorphism in patients with type I diabetes. AB - Our purpose was to investigate a possible relationship between occurrence of type I diabetes and polymorphism of the mannose binding lectin gene. Polymorphism of codon 54 of the mannose binding lectin (MBL) gene, whose presence of the minority allele leads to significant reduction of serum MBL concentration, was investigated in 128 Japanese patients with type I diabetes and 78 healthy volunteers by restriction fragment length polymorphism method. Frequencies of the minority allele were compared between the patient group and the control group. Frequency of the minority allele was 24.2% in the patient group and 19.9% in the control group. The probability of being heterozygous or homozygous for the minority allele was 41.4% in the patient group and 33.3% in the control group. Patients with DRB1*0405-DQB1*0401 and/or DRB1*0901-DQB1*0303 haplotypes, the two major type I diabetes-prone human leukocyte antigen haplotypes, showed a slightly higher probability of being heterozygous or homozygous for allele B of the MBL gene. Possession of the minority allele of the MBL gene may be a minor risk factor for having type I diabetes. PMID- 12770794 TI - Linkage disequilibrium between S65C HFE mutation and HLA A29-B44 haplotype in Terceira Island, Azores. AB - Our objective was to investigate the frequency of HFE gene mutations and to study linkage disequilibrium (LD) between HLA-Class I alleles and these mutations in the population of Terceira Island, Azores, Portugal. A total of 218 unrelated individuals were investigated. Three HFE mutations--C282Y, H63D, and S65C--were identified by restriction endonuclease digestion of polymerase chain reaction (PCR)-amplified genomic DNA. HLA-Class I alleles were typed by PCR-single-strand polymorphism. Gene frequencies and LD were estimated using Arlequin V 1.1. Six genotypes were found in the population: WT/WT (58.3%), H63D/WT (31.2%), H63D/H63D (2.3%), H63D/C282Y (0.9%), S65C/WT (4.1%), and C282Y/WT (3.2%). No cases of C282Y or S65C homozygosity were identified. HLA haplotype A3-B7 was in LD with C282Y; HLA alleles A29, B44, and HLA haplotype A29-B44 were in LD with S65C mutation. HFE gene frequencies in this population are similar to those in other European populations; HFE S65C mutation was found in LD with the alleles A29, B44, and with A29-B44 HLA haplotype. PMID- 12770795 TI - Lack of association of CCR2-64I and CCR5-Delta 32 with type 1 diabetes and latent autoimmune diabetes in adults. AB - It is well known that type 1 diabetes mellitus (T1DM) is a complex genetic disease resulting from the autoimmune destruction of pancreatic beta cells. Several genes have been associated with susceptibility and/or protection for T1DM, but the disease risk is mostly influenced by genes located in the class II region of the major histocompatibility complex. The attraction of leukocytes to tissues is essential for inflammation and the beginning of autoimmune reaction. The process is controlled by chemokines, which are chemotactic cytolines. Some studies have shown that CCR2-64I and CCR5-Delta 32 might be important for protection of susceptibility to some immunologically-mediated disorders. In the present study, we demonstrate the lack of association between CCR2-64I and CCR5 Delta 32 gene polymorphism and TIDM and we describe a new method for a simple and more precise genotyping of the CCR2 gene. PMID- 12770796 TI - No association between the -1031 polymorphism in the TNF-alpha promoter region and type 1 diabetes. AB - Tumor necrosis factor alpha (TNF-alpha) is an important immunomodulator and is believed to be involved in the development or progression of type 1 diabetes. In the following study, we evaluated TNF-alpha promoter polymorphisms at positions 863 and -1031 and their association with type 1 diabetes in a group of 210 diabetic patients from Lebanon. Our results show that in our population, the C allele is predominant at position -863, whereas the A allele is very rare (2%). At position -1031, however, the C and T allele distribution was similar in both the patient (17.8% vs 82.2%, respectively) and the control (21.4% vs 79.6%) groups. No association of TNF-alpha genotype at position 1031 with type 1 diabetes was found as demonstrated by the family-based association test and the transmission disequilibrium test. However, when patient genotypes were compared, the recessive CC genotype was only found in type 1 diabetic males but not in type 1 diabetic females. This observation, however, requires further investigation in a larger sample before conclusive association to gender is suggested. In conclusion, our results demonstrate that no association between TNF-alpha polymorphism and type 1 diabetes seems to exist in our population. PMID- 12770797 TI - HLA class II and TNF genes in African Americans from the Southeastern United States: regional differences in allele frequencies. AB - Knowledge of population major histocompatibility complex gene frequencies is important for construction of organ donor pools and for studies of disease association. Human leukocyte antigen DRB1 (HLA-DRB1), HLA-DQB1, and TNFalpha -308 (G-A) promoter genetic typing was performed in 112 healthy, unrelated African Americans (AAs) from the southeastern United States. Allele frequencies were compared with published frequency data from other AA populations. Our AA population had the highest frequency of HLA- DRB1*09 (6.7%) reported in any AA population. The frequency of the TNF alpha -308A polymorphism was also high (14.4%), when compared with published frequencies in AAs. Significant regional differences in the distribution of most HLA-DRB1 and HLA-DQB1 alleles were observed in all AA populations examined. The AA HLA-DRB1 and -DQB1 frequencies also differed from published Caucasian frequencies. This is the first report describing the distribution of TNF alpha promoter alleles in the Southeastern United States. The high DRB1*09 and TNF alpha -308A allele frequencies of our population most resemble the frequencies of these alleles in certain West African populations. These varying major histocompatibility complex gene frequencies may reflect different regional population structures among AAs in the United States, which may be due to differences in ancestral origins, migration, and racial admixture. PMID- 12770799 TI - Nomenclature for factors of the HLA system, update January 2003. PMID- 12770798 TI - Killer-cell immunoglobulin-like receptor (KIR) nomenclature report, 2002. PMID- 12770801 TI - [Why and how to sensibly apply the perinatal project?]. PMID- 12770800 TI - Nomenclature for factors of the HLA system, update February 2003. PMID- 12770802 TI - [Deep pelvic endometriosis: management and proposal for a "surgical classification"]. AB - Deep pelvic endometriosis presents essentially in the form of a painful syndrome dominated by deep dyspareunia and painful functional symptoms that recur according to the menstrual cycle, with the semiology directly correlated with the location of the lesions (bladder, rectum). It is essential to investigate these deep endometriosis lesions and draw up a precise map, which is the only way to be sure that exeresis will be complete. The treatment of first intention remains surgery, and medical treatment is only palliative in the majority of cases. Success of treatment depends on how radical surgical exeresis is. Based on analysis of the anatomical distribution of deep pelvic endometriosis lesions, a "surgical classification" is proposed with the aim of establishing standard modes for surgical treatment. Further studies are required to clarify the place and modes for pre- and postoperative medical treatment. PMID- 12770804 TI - [Carbon dioxide laser vaporization of genital condyloma in pregnancy]. AB - OBJECTIVE: To evaluate therapeutic effectiveness of the carbon dioxide laser on genital condyloma in pregnancy. PATIENTS AND METHODS: From 1998 to 2002, 18 pregnant women with genital condyloma were treated with laser vaporization. Treatment was given without regard to the gestational period. All the women were treated at least once. Condyloma localisation, treatment gestational age and complication rate were analysed. RESULTS: Patients were treated at a gestational age of 15-38 weeks. During pregnancy follow-up, 2 patients had recurrences that required repeated treatment before delivery. In each case, no condylomas were found during delivery. There were no abortions, no premature birth and no complications (bleeding, infections) in our study. CONCLUSION: Laser vaporisation is a safe and effective treatment for condyloma during pregnancy. PMID- 12770803 TI - [Ovarian drilling for surgical approach of polycystic ovary syndrome]. AB - OBJECTIVES: Show the different surgical approaches for the treatment of polycystic ovarian syndrome (SOPK). PATIENTS AND METHODS: Seventy-one patients treated by ovarian drilling by laparoscopy and 47 by hydrolaparoscopy. The main inclusion criterion was the absence of response after 6 months of stimulation by clomifene citrate for oligo-amenorrheic patients. RESULTS: After a review of surgical series published of ovarian drilling, we found 73% pregnancy rate after laparoscopic treatment and 61 % after hydrolaparoscopy. DISCUSSION AND CONCLUSION: Ovarian drilling appears as an alternative to ovarian hyperstimulation with intra-uterine insemination after failure of clomifene citrate and/or metformine. Surgical treatment decreases multiple pregnancies and miscarriages. Hydrolaparoscopy is more efficient than laparoscopy with less morbidity in particular in obese patients. PMID- 12770805 TI - [Oral contraception in France in 2001: results of an opinion poll survey conducted on 3609 women between 15 and 45]. AB - OBJECTIVES: The aim of this study concerning the taking of the pill in France in 2001 was threefold, i.e. to assess its rate as well as its characteristics of use, and to appraise the most frequent side effects as reported by women. MATERIAL AND METHOD: Three thousand six hundred and nine women representative of the French female population between 15 and 45 years of age were recruited thanks to a survey, which took place in 2001. The data were collected from self questionnaires. RESULTS: Women on the whole have quite a good opinion of oral contraception and most of them are convinced of its efficiency. As far as pill tolerance is concerned, opinions do vary, more than half of the women judging that being on the pill is not without side-effects. Though, the rate of use of oral contraceptives has increased by 12% since 1994. Most women (48%) use first and second generation pills and this in all age brackets. Thirty per cent of women aged 30 to 45 keep loyal to the same patent medicine, which they keep using for more than 10 years. Among the side-effects that can be found, two of them- putting on weight (31%) and hydrosodium retention (26%)--are the most frequently quoted, in all age brackets. This accounts for the relatively low ratio of women who find their pill quite satisfactory (58% of the cases). DISCUSSION AND CONCLUSION: Despite the diversity of all the different patented pills that are available, efforts are still to be made in order to reduce what side-effects are encountered when using them. PMID- 12770806 TI - [Factors associated with weight gain in women using oral contraceptives: results of a French 2001 opinion poll survey conducted on 1665 women]. AB - OBJECTIVES: To study how often women put on weight when on the pill and to analyse the relationship between that gain in weight and the characteristics of the last 2 types of pill that had been used. PATIENTS AND METHODS: Three thousand six hundred and nine women representative of the French female population, aged 15 to 45, were recruited thanks to a survey that took place in 2001. Our study mainly concerned the 1665 women who were actually taking the pill at the time of the survey. The data were collected from self-questionnaires. RESULTS: Thirty per cent of women declared to have gained weight since using their latest pill - only one more kg for 4% of them, 2 kg for 10% but 3 kg or more for the remaining 16%. Gain in weight was more frequent with women less than 25 years of age (35%) than with older ones (29%). This gain in weight did not vary according to either the type of pill, which was then used, or the length of time spent in using it, or the age of first using. It was more frequent when found with other side effects such as breast pain, skin disorders or metrorrhaegias; it was less frequent among women who had already been on the pill in the past than among women using an oral contraceptive for the first time (28% vs 34%; P = 0.008). The shorter the taking the latest pill had been, the greater the frequency of gain in weight was (P = 0.005), women who had presented the most side-effects in the past having changed their pill more rapidly than other women. Finally, a gain in weight was found far more often in women who "did" put on weight with their latest pill than in those who "did not" (53% vs 14%; P = 0.0001). All in all, 8% of women who had been previously been taking the pill had given up this method over a weight problem. DISCUSSION AND CONCLUSION: Putting on weight when on the pill being in the long term independent of the type of patent medicine used, it would seem necessary to orientate new research centred both on a chemical and a biological as well as a nutritional approach, so as to answer one of the major preoccupations of oral contraceptive users to the fullest. PMID- 12770807 TI - [Meigs' syndrome. One case]. AB - Meigs' syndrome is a benign tumor of the ovary with recurrent serous effusion which can resorb however after surgery. It is rarely seen. Its physiopathology remains obscure. Presently, our concern is a 34-year-old woman, admitted in an abdominal tension board, seriously affected by dyspnoea and whose state revealed a malignant process. The pelvic abdominal echography and the thoracic radiography have played a prominent role in the diagnostic approach. Ablating the ovarian tumor leads to a quick recovery. The histological nature of this tumor reveals a huge mucinous cystadenoma. PMID- 12770808 TI - [Pregnancy after embolization of a uterine leiomyomata]. AB - Uterine artery embolization could be useful in the treatment of myoma, either before surgery to facilitate dissection, or totally in place of surgery. Nowadays, this technique seems attractive, considering the development of a conservative and non-invasive medicine. However the question of fertility in women of childbearing-age remains. The case of a pregnancy following uterine artery embolization is here described. All pregnancies from the literature have been reviewed and the discussion includes benefits and drawbacks of this method. PMID- 12770809 TI - [Psychic factors in female unexplained infertility]. AB - Unexplained cases of female infertility were studied from a psychoanalytical point of view emphasizing the unconscious secondary gain from the illness. Three groups of cases were isolated: post traumatic infertility, neurotic infertility and infertility correlated with eating disorders and disturbed self image. Joint consultations between gynecologists and psychoanalysts are proposed. PMID- 12770810 TI - [Evidence-based medicine: principles and applications to obstetrics and to gynecologic surgery]. AB - Evidence-based medicine is a concept developed by David Sackett and colleagues since 1980 at the Medical University center MacMaster, Hamilton city, Ontario. The main idea was to propose an evaluating tool of medical practice allowing a structured way of reasoning and the elimination of self-made opinion. We present a short explanation of evidence-based medicine and compare our daily practice in obstetrics and gynecologic surgery with this new practice paradigm. PMID- 12770811 TI - [Breast biopsies: stereotactic vacuum-assisted core biopsy and stereotactic surgical breast biopsy]. AB - Stereotactically-guided procedures for diagnosis of breast lesions can avoid a lot of surgical biopsies. Stereotactic guidance is used for vacuum-assisted core biopsies and for stereotactic breast biopsies. Technical details of the procedures are described, and the benefits and the limits of these methods are discussed. Indications for breast sampling are proposed according to the Breast Imaging Reporting and Data System (BI-RADS) assessment categories. PMID- 12770812 TI - [Should infertile women with polycystic ovarian syndrome be treated with metformine?]. AB - Polycystic ovary syndrome is a frequent endocrine disorder often associated with insulin resistance and hyperinsulinaemia which may play a role in hyperandrogenism and anovulation. The use of "insulin sensitizing" agents has been suggested to reduce insulin resistance and hyperandrogenism. In that respect, the use of metformin in polycystic ovary syndrome is reviewed. Although its mechanism of action is still unclear, metformin proved to be effective to restore cyclicity and spontaneous ovulation. The synergistic effect of clomiphene citrate and metformin was demonstrated in some studies, suggesting that metformin could be helpful for women with clomiphene citrate resistance. However, the potential effect of metformin administration for reducing hyperstimulation in women treated with exogenous FSH, or for preventing early miscarriages has to be confirmed. Here, we propose a guideline for the use of metformin, as an adjuvant therapy, to restore cyclicity and ovulation in women with polycystic ovary syndrome. PMID- 12770813 TI - [Metaplasia and high grade CIN. Diagnostic difficulties. Gynecol Obstet Fertil 2002; 30: 845-849]. PMID- 12770814 TI - [Metaplasia and high grade CIN. Diagnostic difficulties. Gynecol Obstet Fertil 2002; 30: 845-849]. PMID- 12770815 TI - [Metaplasia and high grade CIN. Diagnostic difficulties. Gynecol Obstet Fertil 2002; 30: 845-849]. PMID- 12770816 TI - [Prolapse in the young woman: study of risk factors. Gynecol Obstet Fertil 2002; 30:673-6]. PMID- 12770817 TI - [Prolapse in the young woman: study of risk factors. Gynecol Obstet Fertil 2002; 30:673-6]. PMID- 12770818 TI - Neurocrine Biosciences, Inc. Soporific science. PMID- 12770819 TI - Understanding HTLV-I protease. PMID- 12770821 TI - Multidimensional chemical genetic analysis of diversity-oriented synthesis derived deacetylase inhibitors using cell-based assays. AB - Systematic chemical genetics aims to explore the space representing interactions between small molecules and biological systems. Beyond measuring binding interactions and enzyme inhibition, measuring changes in the activity of proteins in intact signaling networks is necessary. Toward this end, we are partitioning chemical space into regions with different biological activities using a panel of cell-based assays and small molecule "chemical genetic modifiers." Herein, we report on the use of this methodology for the discovery of 617 small molecule inhibitors of histone deacetylases from a multidimensional screen of an encoded, diversity-oriented synthesis library. Following decoding of chemical tags and resynthesis, we demonstrate the selectivity of one inhibitory molecule (tubacin) toward alpha-tubulin deacetylation and another (histacin) toward histone deacetylation. These small molecules will facilitate dissecting the role of acetylation in a variety of cell biological processes. PMID- 12770822 TI - Chemical genetic modifier screens: small molecule trichostatin suppressors as probes of intracellular histone and tubulin acetylation. AB - Histone deacetylase (HDAC) inhibitors are being developed as new clinical agents in cancer therapy, in part because they interrupt cell cycle progression in transformed cell lines. To examine cell cycle arrest induced by HDAC inhibitor trichostatin A (TSA), a cytoblot cell-based screen was used to identify small molecule suppressors of this process. TSA suppressors (ITSAs) counteract TSA induced cell cycle arrest, histone acetylation, and transcriptional activation. Hydroxamic acid-based HDAC inhibitors like TSA and suberoylanilide hydroxamic acid (SAHA) promote acetylation of cytoplasmic alpha-tubulin as well as histones, a modification also suppressed by ITSAs. Although tubulin acetylation appears irrelevant to cell cycle progression and transcription, it may play a role in other cellular processes. Small molecule suppressors such as the ITSAs, available from chemical genetic suppressor screens, may prove to be valuable probes of many biological processes. PMID- 12770823 TI - Rational design and molecular diversity for the construction of anti-alpha bungarotoxin antidotes with high affinity and in vivo efficiency. AB - The structure of peptide p6.7, a mimotope of the nicotinic receptor ligand site that binds alpha-bungarotoxin and neutralizes its toxicity, was compared to that of the acetylcholine binding protein. The central loop of p6.7, when complexed with alpha-bungarotoxin, fits the structure of the acetylcholine binding protein (AChBP) ligand site, whereas peptide terminal residues seem to be less involved in toxin binding. The minimal binding sequence of p6.7 was confirmed experimentally by synthesis of progressively deleted peptides. Affinity maturation was then achieved by random addition of residues flanking the minimal binding sequence and by selection of new alpha-bungarotoxin binding peptides on the basis of their dissociation kinetic rate. The tetra-branched forms of the resulting high-affinity peptides were effective as antidotes in vivo at a significantly lower dose than the tetra-branched lead peptide. PMID- 12770824 TI - Characterization of the mupirocin biosynthesis gene cluster from Pseudomonas fluorescens NCIMB 10586. AB - The polyketide antibiotic mupirocin (pseudomonic acid) produced by Pseudomonas fluorescens NCIMB 10586 competitively inhibits bacterial isoleucyl-tRNA synthase and is useful in controlling Staphylococcus aureus, particularly methicillin resistant Staphylococcus aureus. The 74 kb mupirocin biosynthesis cluster has been sequenced, and putative enzymatic functions of many of the open reading frames (ORFs) have been identified. The mupirocin cluster is a combination of six larger ORFs (mmpA-F), containing several domains resembling the multifunctional proteins of polyketide synthase and fatty acid synthase type I systems, and individual genes (mupA-X and macpA-E), some of which show similarity to type II systems (mupB, mupD, mupG, and mupS). Gene knockout experiments demonstrated the importance of regions in mupirocin production, and complementation of the disrupted gene confirmed that the phenotypes were not due to polar effects. A model for mupirocin biosynthesis is presented based on the sequence and biochemical evidence. PMID- 12770825 TI - A complete gene cluster from Streptomyces nanchangensis NS3226 encoding biosynthesis of the polyether ionophore nanchangmycin. AB - The PKS genes for biosynthesis of the polyether nanchangmycin are organized to encode two sets of proteins (six and seven ORFs, respectively), but are separated by independent ORFs that encode an epimerase, epoxidase, and epoxide hydrolase, and, notably, an independent ACP. One of the PKS modules lacks a corresponding ACP. We propose that the process of oxidative cyclization to form the polyether structure occurs when the polyketide chain is still anchored on the independent ACP before release. 4-O-methyl-L-rhodinose biosynthesis and its transglycosylation involve four putative genes, and regulation of nanchangmycin biosynthesis seems to involve activation as well as repression. In-frame deletion of a KR6 domain generated the nanchangmycin aglycone with loss of 4-O-methyl-L rhodinose and antibacterial activity, in agreement with the assignments of the PKS domains catalyzing specific biosynthetic steps. PMID- 12770826 TI - Synthetic inhibitors of proline-rich ligand-mediated protein-protein interaction: potent analogs of UCS15A. AB - The proline-rich motif in proteins is known to function as a ligand sequence that binds to protein modules such as SH3, WW, and several other protein interaction domains. These proline-rich ligand-mediated protein-protein interactions (abbreviated PLPI) are important in many signaling pathways that are involved in various diseases. Our previous studies showed that UCS15A, produced by Streptomyces species, inhibited PLPI. Here we report on synthetic analogs of UCS15A that show more potent activity than UCS15A in inhibiting PLPI. A synthetic analog, compound 2c, blocked in vitro PLPI of Sam68-Fyn-SH3 as well as in vivo PLPI of Grb2-Sam68 and Grb2-Sos1. Activation of MEK was also inhibited by compound 2c. Unlike UCS15A, compound 2c was an order of magnitude less cytotoxic and did not cause morphological changes in treated cells. PMID- 12770827 TI - Biosynthesis of structurally novel carotenoids in Escherichia coli. AB - Previously, we utilized in vitro evolution to alter the catalytic functions of several carotenoid enzymes and produce the novel carotenoids tetradehydrolycopene and torulene in Escherichia coli. Here we report on the successful extension of these pathways and the C(30) carotenoid diaponeurosporene pathway with additional carotenoid genes. Extension of the known acyclic C(30) pathway with C(40) carotenoid enzymes-spheroidene monooxygenase and lycopene cyclase-yielded new oxygenated acylic products and the unnatural cyclic C(30) diapotorulene, respectively. Extension of acyclic C(40) pathways with spheroidene monooxygenase generated novel oxygenated carotenoids including the violet phillipsiaxanthin. Extension of the torulene biosynthetic pathway with carotene hydroxylase, desaturase, glucosylase, and ketolase yielded new torulene derivatives. These results demonstrate the utility of extending an in vitro evolved central metabolic pathway with catalytically promiscuous downstream enzymes in order to generate structurally novel compounds. PMID- 12770829 TI - Is the Atkins diet on to something? No, it's not a healthy way to eat. But the high-protein, low-carbohydrate diet may hold a few important lessons about weight loss and healthy eating. PMID- 12770831 TI - The latest on macular degeneration. Macular degeneration is the main cause of severe vision loss in older people. Prevention is proving elusive, but new treatments hold promise. PMID- 12770830 TI - Health tips for air travelers. Jet lag, cramped seats, bad air--flying can make you wish you'd never left home. Here are some suggestions for making air travel easier and perhaps a bit healthier. PMID- 12770828 TI - Mu transpososome architecture ensures that unfolding by ClpX or proteolysis by ClpXP remodels but does not destroy the complex. AB - The Clp/Hsp100 ATPases are protein unfoldases that both alter protein conformation and target proteins for degradation. An unresolved question has been how such seemingly destructive enzymes can "remodel" some protein substrates rather than destroy them. Here, we investigate the products of ClpX-mediated remodeling of a hyper-stable protein-DNA complex, the Mu transpososome. We find that although an oligomeric complex is maintained, release of some subunits accompanies ClpX action. Replacement of transposase's endogenous ClpX-recognition sequence with an exogenous signal reveals that the mechanism of remodeling is independent of both the recognition signal and the identity of the unfoldase. Finally, examination of the transposase-DNA contacts reveals only a localized region that is altered during remodeling. These results provide a framework for protein remodeling, wherein the physical attributes of a complex can limit the unfolding activity of its remodeler. PMID- 12770833 TI - Are you eating the right kind of breakfast cereal? PMID- 12770832 TI - What really causes heart attacks. In most cases, it's not a plugged artery, according to the "vulnerable plaque" theory. But searching for the vulnerable plaque may also be a mistake. PMID- 12770834 TI - Blood donation linked to restless legs syndrome. PMID- 12770835 TI - Calcium-channel blockers may prevent dementia. PMID- 12770836 TI - By the way, doctor. The Harvard Health Letter seems pretty dubious about the PSA test for prostate cancer. But a PSA test seemed to catch Senator John Kerry's cancer early, so he got surgery that may have cured him. I just don't get why you have doubts about testing and early detection. PMID- 12770837 TI - By the way, doctor. At a hospital near my home, they're using something called endoluminal grafts to fix abdominal aortic aneurysms. They're claiming that you're out of the hospital in one or two days and back on the golf course in a month. Too good to be true? PMID- 12770838 TI - Selective blockade of lysophosphatidic acid LPA3 receptors reduces murine renal ischemia-reperfusion injury. AB - Lysophosphatidic acid (LPA) released during ischemia has diverse physiological effects via its G protein-coupled receptors, LPA1, LPA2, and LPA3 (formerly Edg 2, -4, and -7). We tested the hypothesis that selective blockade of LPA receptors affords protection from renal ischemia-reperfusion (I/R) injury. By real-time PCR, LPA1-3 receptor mRNAs were expressed in mouse renal cortex, outer medulla, and inner medulla with the following rank order LPA3 = LPA2 > LPA1. In C57BL/6 mice whose kidneys were subjected to ischemia and reperfusion, treatment with a selective LPA3 agonist, oleoyl-methoxy phosphothionate (OMPT), enhanced injury. In contrast, a dual LPA1/LPA3-receptor antagonist, VPC-12249, reduced I/R injury, but this protective effect was lost when the antagonist was coadministered with OMPT. Interestingly, delaying administration of VPC-12249 until 30 min after the start of reperfusion did not alter its efficacy significantly. We conclude that VPC-12249 reduces renal I/R injury predominantly by LPA3 receptor blockade and could serve as a novel compound in the treatment of ischemia acute renal failure. PMID- 12770839 TI - Multiple epithelial Na+ channel domains participate in subunit assembly. AB - Epithelial sodium channels (ENaCs) are composed of three structurally related subunits that form a tetrameric channel. The Xenopus laevis oocyte expression system was used to identify regions within the ENaC alpha-subunit that confer a dominant negative phenotype on functional expression of alphabetagamma-ENaC to define domains that have a role in subunit-subunit interactions. Coexpression of full-length mouse alphabetagamma-ENaC with either 1) the alpha-subunit first membrane-spanning domain and short downstream hydrophobic domain (alpha-M1H1); 2) alpha-M1H1 and its downstream hydrophilic extracellular loop (alpha-M1H1-ECL); 3) the membrane-spanning domain of a control type 2 transmembrane protein (glutamyl transpeptidase; gamma-GT) fused to the alpha-ECL (gamma-GT-alpha-ECL); 4) the extracellular domain of a control type 1 transmembrane protein (Tac) fused to the alpha-subunit second membrane-spanning domain and short upstream hydrophobic domain (Tac-alpha-H2M2); or 5) the alpha-subunit cytoplasmic COOH terminus (alpha Ct) significantly reduced amiloride-sensitive Na+ currents in X. laevis oocytes. Functional expression of Na+ channels was not inhibited when full-length alphabetagamma-ENaC was coexpressed with either 1) the alpha-ECL lacking a signal anchor sequence, 2) alpha-M1H1 and alpha-Ct expressed as a fusion protein, 3) full-length gamma-GT, or 4) full-length Tac. Furthermore, the expression of ROMK channels was not inhibited when full-length ROMK was coexpressed with either alpha-M1H1-ECL or alpha-Ct. Full-length FLAG-tagged alpha-, beta-, or gamma-ENaC coimmunoprecipitated with myc-tagged alpha-M1H1-ECL, whereas wild-type gamma-GT did not. These data suggest that multiple sites within the alpha-subunit participate in subunit-subunit interactions that are required for proper assembly of the heterooligomeric ENaC complex. PMID- 12770840 TI - Early transcriptional effects of aldosterone in a mouse inner medullary collecting duct cell line. AB - The mineralocorticoid aldosterone is a major regulator of Na+ and acid-base balance and control of blood pressure. Although the long-term effects of aldosterone have been extensively studied, the early aldosterone-responsive genes remain largely unknown. Using DNA array technology, we have characterized changes in gene expression after 1 h of exposure to aldosterone in a mouse inner medullary collecting duct cell line, mIMCD-3. Results from three independent microarray experiments revealed that the expression of many transcripts was affected by aldosterone treatment. Northern blot analysis confirmed the upregulation of four distinct transcripts identified by the microarray analysis, namely, the serum and glucose-regulated kinase sgk, connective tissue growth factor, period homolog, and preproendothelin. Immunoblot analysis for preproendothelin demonstrated increased protein expression. Following the levels of the four transcripts over time showed that each had a unique pattern of expression, suggesting that the cellular response to aldosterone is complex. The results presented here represent a novel list of early aldosterone-responsive transcripts and provide new avenues for elucidating the mechanism of acute aldosterone action in the kidney. PMID- 12770841 TI - Hydrostatic pressure-regulated ion transport in bladder uroepithelium. AB - The effect of hydrostatic pressure on ion transport in the bladder uroepithelium was investigated. Isolated rabbit uroepithelium was mounted in modified Ussing chambers and mechanically stimulated by applying hydrostatic pressure across the mucosa. Increased hydrostatic pressure led to increased mucosal-to-serosal Na+ absorption across the uroepithelium via the amiloride-sensitive epithelial Na+ channel. In addition to this previously characterized pathway for Na+ absorption, hydrostatic pressure also induced the secretion of Cl- and K+ into the mucosal bathing solution under short-circuit conditions, which was confirmed by a net serosal-to-mucosal flux of 36Cl- and 86Rb+. K+ secretion was likely via a stretch activated nonselective cation channel sensitive to 100 microM amiloride, 10 mM tetraethylammonium, 3 mM Ba2+, and 1 mM Gd3+. Hydrostatic pressure-induced ion transport in the uroepithelium may play important roles in electrolyte homeostasis, volume regulation, and mechanosensory transduction. PMID- 12770842 TI - Effects of ultraviolet-B radiation on cotton (Gossypium hirsutum L.) morphology and anatomy. AB - Cotton (Gossypium hirsutum L.) crop, cultivated between 40 degrees N and 40 degrees S, is currently experiencing 2-11 kJ m-2 d-1 of UV-B radiation. This is predicted to increase in the near future. An experiment was conducted to study the effect of enhanced UV-B radiation on vegetative and reproductive morphology and leaf anatomy of cotton in sunlit, controlled environment chambers. From emergence to harvest, cotton plants were exposed to 0, 8 or 16 kJ m-2 d-1 of UV-B in a square wave approach for 8 h from 0800 to 1600 h. Changes in plant height, internode and branch length, mainstem node number, leaf area, length and area of petals and bracts, and anther number per flower were recorded. Epidermal cell and stomatal density, stomatal index, leaf thickness, and epidermal, palisade and mesophyll tissue thickness were also measured. Initial chlorotic symptoms on leaves turned into necrotic patches on continued exposure to enhanced UV-B. Exposure to high UV-B reduced both vegetative and reproductive parameters and resulted in a smaller canopy indicating sensitivity of cotton to UV-B radiation. Enhanced UV-B radiation increased epicuticular wax content on adaxial leaf surfaces, and stomatal index on both adaxial and abaxial leaf surfaces. Leaf thickness was reduced following exposure to UV-B owing to a decrease in thickness of both the palisade and mesophyll tissue, while the epidermal thickness remained unchanged. The vegetative parameters studied were affected only by high levels of UV-B (16 kJ m-2 d-1), whereas the reproductive parameters were reduced at both ambient (8 kJ m-2 d-1) and high UV-B levels. The study shows that cotton plants are sensitive to UV-B at both the whole plant and anatomical level. PMID- 12770843 TI - Comparative wood anatomy of epacrids (Styphelioideae, Ericaceae s.L.). AB - The wood anatomy of 16 of the 37 genera within the epacrids (Styphelioideae, Ericaceae s.l.) is investigated by light and scanning electron microscopy. Several features in the secondary xylem occur consistently at the tribal level: arrangement of vessel-ray pits, distribution of axial parenchyma, ray width, and the presence and location of crystals. The primitive nature of Prionoteae and Archerieae is supported by the presence of scalariform perforation plates with many bars and scalariform to opposite vessel pitting. The wood structure of Oligarrheneae is similar to that of Styphelieae, but the very narrow vessel elements, exclusively uniseriate rays and the lack of prismatic crystals in Oligarrheneae distinguish these two tribes. The secondary xylem of Monotoca tamariscina indicates that it does not fit in Styphelieae; a position within Oligarrheneae is possible. Like most Cosmelieae, all Richeeae are characterized by exclusively scalariform perforation plates with many bars, a very high vessel density and paratracheal parenchyma, although they clearly differ in ray width (exclusively uniseriate rays in Cosmelieae vs. uniseriate and wide multiseriate rays in Richeeae). Several wood anatomical features confirm the inclusion of epacrids in Ericaceae s.l. Furthermore, there are significant ecological implications. The small vessel diameter and high vessel frequency in many epacrids are indicative of a high conductive safety to avoid embolism caused by freeze-thaw cycles, while the replacement of scalariform by simple vessel perforation plates and an increase in vessel diameter would suggest an increased conductive efficiency, which is especially found in mesic temperate or tropical Styphelieae. PMID- 12770844 TI - Estimating photosynthetic radiation use efficiency using incident light and photosynthesis of individual leaves. AB - It has been theorized that photosynthetic radiation use efficiency (PhRUE) over the course of a day is constant for leaves throughout a canopy if leaf nitrogen content and photosynthetic properties are adapted to local light so that canopy photosynthesis over a day is optimized. To test this hypothesis, 'daily' photosynthesis of individual leaves of Solanum melongena plants was calculated from instantaneous rates of photosynthesis integrated over the daylight hours. Instantaneous photosynthesis was estimated from the photosynthetic responses to photosynthetically active radiation (PAR) and from the incident PAR measured on individual leaves during clear and overcast days. Plants were grown with either abundant or scarce N fertilization. Both net and gross daily photosynthesis of leaves were linearly related to daily incident PAR exposure of individual leaves, which implies constant PhRUE over a day throughout the canopy. The slope of these relationships (i.e. PhRUE) increased with N fertilization. When the relationship was calculated for hourly instead of daily periods, the regressions were curvilinear, implying that PhRUE changed with time of the day and incident radiation. Thus, linearity (i.e. constant PhRUE) was achieved only when data were integrated over the entire day. Using average PAR in place of instantaneous incident PAR increased the slope of the relationship between daily photosynthesis and incident PAR of individual leaves, and the regression became curvilinear. The slope of the relationship between daily gross photosynthesis and incident PAR of individual leaves increased for an overcast compared with a clear day, but the slope remained constant for net photosynthesis. This suggests that net PhRUE of all leaves (and thus of the whole canopy) may be constant when integrated over a day, not only when the incident PAR changes with depth in the canopy, but also when it varies on the same leaf owing to changes in daily incident PAR above the canopy. The slope of the relationship between daily net photosynthesis and incident PAR was also estimated from the photosynthetic light response curve of a leaf at the top of the canopy and from the incident PAR above the canopy, in place of that measured on individual leaves. The slope (i.e. net PhRUE) calculated in this simple way did not differ statistically from that calculated using data from individual leaves. PMID- 12770845 TI - Nutrient dynamics throughout the rotation of Eucalyptus clonal stands in Congo. AB - The dynamics of the main nutrient fluxes of the biological cycle were quantified in a clonal Eucalyptus plantation throughout the whole planted crop rotation: current annual requirements of nutrients, uptake from the soil, internal translocations within trees, return to soil (litterfall and crown leaching) and decomposition in the forest floor. As reported for other species, two growth periods were identified in these short-rotation plantations: (1) a juvenile phase up to canopy closure, during which the uptake of nutrients from the soil reserves supplied most of the current requirements; and (2) a second phase up to harvest, characterized by intense nutrient recycling processes. Internal translocation within trees supplied about 30 % of the annual requirements of N and P from 2 years of age onwards, and about 50 % of the K requirement. The mineralization of large amounts of organic matter returned to the soil with litterfall during stand development represented a key process providing nutrients to the stand at the end of the rotation. The importance of the recycling processes was clearly shown by the small amounts of nutrients permanently immobilized in the ligneous components of trees, compared with the total requirements accumulated over the stand rotation which were two to four times higher. Small pools of nutrients circulating quickly in the ecosystem made it possible to produce high amounts of biomass in poor soils. The sustainability of these plantations will require fertilizer inputs that match the changes in soil fertility over successive rotations, mainly linked to the dynamics of organic matter in this tropical soil. PMID- 12770846 TI - Exogenously applied jasmonic acid induces changes in apical meristem morphology of potato stolons. AB - Hooked apex stolons and initial swelling stolons of potato plants were treated with 3 x 10-8 mol l-1 jasmonic acid (JA) to study the effect of this compound on histology, cell expansion and tissue differentiation. In hooked apex stolons, JA application increased the meristem thickness and reduced the length of the leaf primordia, whereas in initial swelling stolons narrowing of the apical region, absence of leaf primordia and swelling of the subapical meristem were evident. Early vascular tissue differentiation was observed in response to JA treatment, especially of xylem elements from regions proximal to the tunic. Protoxylem elements, such as tracheal elements, were present with thin primary cell walls. The cell area was measured in two zones: zone I, central mother cells situated immediately under the tunic; and zone II, rib meristem cells. JA caused a four- and six-fold increase in cell area in both zones in hooked apex stolons and initial swelling stolons, respectively. Thus, tuber formation is concluded to occur as a consequence of increased cell expansion, a reduction in the length of leaf primordia, enlargement of meristems, and early vascular tissue differentiation. PMID- 12770847 TI - Karyotype analysis of four Vicia species using in situ hybridization with repetitive sequences. AB - Mitotic chromosomes of four Vicia species (V. sativa, V. grandiflora, V. pannonica and V. narbonensis) were subjected to in situ hybridization with probes derived from conserved plant repetitive DNA sequences (18S-25S and 5S rDNA, telomeres) and genus-specific satellite repeats (VicTR-A and VicTR-B). Numbers and positions of hybridization signals provided cytogenetic landmarks suitable for unambiguous identification of all chromosomes, and establishment of the karyotypes. The VicTR-A and -B sequences, in particular, produced highly informative banding patterns that alone were sufficient for discrimination of all chromosomes. However, these patterns were not conserved among species and thus could not be employed for identification of homologous chromosomes. This fact, together with observed variations in positions and numbers of rDNA loci, suggests considerable divergence between karyotypes of the species studied. PMID- 12770849 TI - Tuberculosis chemotherapy: still a double-edged sword. PMID- 12770848 TI - Comparative developmental anatomy of the root in three species of Cladopus (Podostemaceae). AB - Root meristem structure and root branching in three species of Cladopus were investigated from developmental and anatomical perspectives. Cladopus fukiensis has a compressed bell-shaped meristem at the apex of a compressed subcylindrical root, while C. javanicus and perhaps C. nymanii, with a ribbon-like root, have a half lozenge-shaped ( subset as seen from above) meristem composed of an apical meristem of cubic cells and a marginal meristem of rectangular cells. The dorsiventrality of the meristem results in root dorsiventrality, and a marginal meristem contributes to the broadening of the root. Comparisons of meristem structure and root morphology suggest that the ribbon-like root of, e.g. C. javanicus, evolved towards the foliose root of Hydrobryum, sister to the genus Cladopus, by loss of an indeterminate apical meristem. The lateral root of C. javanicus initiates within the meristem of a parent root. The dorsal dermal layer and inner cells of the lateral-root meristem appear endogenously under the dermal layer of the parent root, while the ventral layer is derived exogenously from a ventral dermal layer continuous with the parent-root meristem. This mosaic pattern of exogenous and endogenous root formation differs from the truly exogenous formation seen in Hydrobryum and Zeylanidium. The dorsiventral mosaic origin of the root meristem may account for root cap asymmetry. PMID- 12770850 TI - New insights from an old model. PMID- 12770851 TI - Inhaled nitric oxide after lung transplantation: no more cosmesis? PMID- 12770852 TI - The blood lung function test: does it exist for asthma? PMID- 12770853 TI - On the physiologic and clinical relevance of lung-borne cytokines during ventilator-induced lung injury. PMID- 12770854 TI - A randomized trial of inhaled nitric oxide to prevent ischemia-reperfusion injury after lung transplantation. AB - Inhalation of nitric oxide (NO) has been advocated as a method to prevent ischemia-reperfusion injury after lung transplantation. We enrolled 84 patients into a concealed, randomized, placebo-controlled trial to evaluate the effect of inhaled NO (20 ppm NO or nitrogen) initiated 10 minutes after reperfusion on outcomes after lung transplantation. The groups (n = 42) were balanced with respect to age, sex, lung disease, procedure, and total ischemic times. PaO2/FIO2 ratios were similar on admission to the intensive care unit (ICU) (NO 361 +/- 134; control patients 357 +/- 132), and over the duration of the study. There were no differences in hemodynamics between the two groups. Severe reperfusion injury (PaO2/FIO2 < 150) was present at the time of admission to the ICU in 14.6% NO patients versus 9.5% of control patients (p = 0.48). The groups had similar median times to first successful trial of unassisted breathing (25 vs. 27 hours; p = 0.76), successful extubation (32 vs. 34 hours; p = 0.65), ICU discharge (3.0 days for both groups), and hospital discharge (27 vs. 29 days; p = 0.563). Five NO versus six control patients died during their hospital stay. Adjusting for age, sex, lung disease etiology, presence of pulmonary hypertension, and total ischemic time did not alter these results. In conclusion, we did not detect a significant effect of inhaled NO administered 10 minutes after reperfusion on physiologic variables or outcomes in lung transplant patients. PMID- 12770856 TI - Recruitment maneuvers in ARDS. PMID- 12770855 TI - Fully automated assessment of inflammatory cell counts and cytokine expression in bronchial tissue. AB - Automated image analysis of bronchial tissue offers the opportunity to quantify stained area and staining intensity in a standardized way to obtain robust estimates of inflammatory cell counts and cytokine expression from multiple large areas of histopathologic sections. We compared fully automated digital image analysis with interactive digital cell counting and semiquantitative scoring of cytokine expression in terms of repeatability and agreement in bronchial biopsies in 52 patients with mild to moderate atopic asthma. Immunohistochemistry with antibodies against CD3, interleukin (IL)-4, IL-5, and interferon-gamma protein was performed on frozen tissue sections, using 3-amino-9-ethylcarbazole as chromogen and hematoxylin as counterstaining. IL-4 and IL-5 messenger RNAs were localized by in situ hybridization without hematoxylin staining. Separation of 3 amino-9-ethylcarbazole and hematoxylin-stained pixels was achieved by linear combination of red- and blue-filtered gray-scale images. Using baseline biopsy specimens, fully automated CD3+ cell counts showed perfect repeatability (r = 1.0) and a strong linear relationship with the interactive procedure (r = 0.98). Automated densitometry showed perfect repeatability (1.0) and a moderate to strong relationship with semiquantitative scoring of protein and messenger RNA expression (r = 0.43-0.89). Relationships between automated and semiquantitative assessments of changes in cytokine expression during 2 years of follow-up were moderate to strong (r = 0.40-0.84). We conclude that fully automated cell counts and automated densitometric analyses in bronchial tissue of patients with asthma are unbiased and help to reduce variability in inflammatory outcomes. PMID- 12770857 TI - Right level of positive end-expiratory pressure in acute respiratory distress syndrome. PMID- 12770858 TI - Mutations of CFTR gene and intermediate sweat chloride levels. PMID- 12770859 TI - Prenatal antibiotic exposure and subsequent atopy. PMID- 12770860 TI - Formoterol and ipratropium in COPD. PMID- 12770861 TI - Revealing the properties of dendritic voltage-gated channels: a new approach to the space clamp problem. PMID- 12770862 TI - Quantitative parsing of cell multi-tasking in wound repair and tissue morphogenesis. PMID- 12770863 TI - Connective tissue polarity unraveled by a markov-chain mechanism of collagen fibril segment self-assembly. AB - The well-established occurrence of pyroelectricity (Lang, 1966) in tissues of living organisms has found a first explanation by a Markov-chain mechanism taking place during collagen fibril self-assembly in extracytoplasmic channels. Recently reported biochemical findings on the longitudinal fusion reactivity of small fibril segments (which undergo C-, N- and C-, C- but not N-, N-terminal fusions; see Graham et al., 2000; Kadler et al., 1996) may provide a mechanism by which a difference in the fusion probabilities P(CC), P(NN) drives the self-assembly into partial macroscopic polar order. In principle, a Markov-chain growth process can lower the noncentrosymmetric infinity 2 symmetry describing dielectric properties of a growing limb (as managed by fibroblasts) into the polar infinity group. It is proposed that macroscopically polar properties enter the biological world by a stochastic mechanism of unidirectional growth. Polarity formation in organisms shows similarity to effects reported for molecular crystals (Hulliger et al., 2002). PMID- 12770864 TI - Correction of conductance measurements in non-space-clamped structures: 1. Voltage-gated K+ channels. AB - To understand functions of a single neuron, such as propagation and generation of synaptic or action potentials, a detailed description of the kinetics and distribution of the underlying ionic conductances is essential. In voltage-clamp experiments, incomplete space clamp distorts the recorded currents, rendering accurate analysis impossible. Here, we present a simple numerical algorithm that corrects such distortions. The method performs a stepwise approximation of the conductance density at the site of a local voltage clamp. This is achieved by estimating membrane conductances in a simulation that yields simulated clamp currents, which are then fitted to the distorted recordings from the non-space clamped structure, relying on accurately reconstructed cell morphology and experimentally determined passive properties. The method enabled accurate retrieval of the local densities, kinetics, and density gradients of somatic and dendritic channels. Neither the addition of noise nor variation of passive parameters significantly reduced the performance of the correction algorithm. The correction method was applied to two-electrode voltage-clamp recordings of K(+) currents from the apical dendrite of layer 5 neocortical pyramidal neurons. The generality and robustness of the algorithm make it a useful tool for voltage clamp analysis of voltage-gated currents in structures of any morphology that is amenable to the voltage-clamp technique. PMID- 12770865 TI - Mechanisms of microtubule-based kinetochore positioning in the yeast metaphase spindle. AB - It has been hypothesized that spatial gradients in kMT dynamic instability facilitate mitotic spindle formation and chromosome movement. To test this hypothesis requires the analysis of kMT dynamics, which have not been resolved at the single kMT level in living cells. The budding yeast spindle offers an attractive system in which to study kMT dynamics because, in contrast to animal cells, there is only one kMT per kinetochore. To visualize metaphase kMT plus-end dynamics in yeast, a strain containing a green fluorescent protein fusion to the kinetochore protein, Cse4, was imaged by fluorescence microscopy. Although individual kinetochores were not resolvable, we found that models of kMT dynamics could be evaluated by simulating the stochastic kMT dynamics and then simulating the fluorescence imaging of kMT plus-end-associated kinetochores. Statistical comparison of model-predicted images to experimentally observed images demonstrated that a pure dynamic instability model for kMT dynamics in the yeast metaphase spindle was unacceptable. However, when a temporally stable spatial gradient in the catastrophe or rescue frequency was added to the model, there was reasonable agreement between the model and the experiment. These results provide the first evidence of temporally stable spatial gradients of kMT catastrophe and/or rescue frequency in living cells. PMID- 12770867 TI - Non-Watson-Crick basepairing and hydration in RNA motifs: molecular dynamics of 5S rRNA loop E. AB - Explicit solvent and counterion molecular dynamics simulations have been carried out for a total of >80 ns on the bacterial and spinach chloroplast 5S rRNA Loop E motifs. The Loop E sequences form unique duplex architectures composed of seven consecutive non-Watson-Crick basepairs. The starting structure of spinach chloroplast Loop E was modeled using isostericity principles, and the simulations refined the geometries of the three non-Watson-Crick basepairs that differ from the consensus bacterial sequence. The deep groove of Loop E motifs provides unique sites for cation binding. Binding of Mg(2+) rigidifies Loop E and stabilizes its major groove at an intermediate width. In the absence of Mg(2+), the Loop E motifs show an unprecedented degree of inner-shell binding of monovalent cations that, in contrast to Mg(2+), penetrate into the most negative regions inside the deep groove. The spinach chloroplast Loop E shows a marked tendency to compress its deep groove compared with the bacterial consensus. Structures with a narrow deep groove essentially collapse around a string of Na(+) cations with long coordination times. The Loop E non-Watson-Crick basepairing is complemented by highly specific hydration sites ranging from water bridges to hydration pockets hosting 2 to 3 long-residing waters. The ordered hydration is intimately connected with RNA local conformational variations. PMID- 12770866 TI - Steered molecular dynamics simulation on the binding of NNRTI to HIV-1 RT. AB - HIV-1 reverse transcriptase (RT) is the primary target for anti-AIDS chemotherapy. Nonnucleoside RT inhibitors (NNRTIs) are very potent and most promising anti-AIDS drugs that specifically inhibit HIV-1 RT. The binding and unbinding processes of alpha-APA, an NNRTI, have been studied using nanosecond conventional molecular dynamics and steered molecular dynamics simulations. The simulation results show that the unbinding process of alpha-APA consists of three phases based on the position of alpha-APA in relation to the entrance of the binding pocket. When alpha-APA is bound in the binding pocket, the hydrophobic interactions between HIV-1 RT and alpha-APA dominate the binding; however, the hydrophilic interactions (both direct and water-bridged hydrogen bonds) also contribute to the stabilizing forces. Whereas Tyr-181 makes significant hydrophobic interactions with alpha-APA, Tyr-188 forms a strong hydrogen bond with the acylamino group (N14) of alpha-APA. These two residues have very flexible side chains and appear to act as two "flexible clamps" discouraging alpha-APA to dissociate from the binding pocket. At the pocket entrance, two relatively inflexible residues, Val-179 and Leu-100, gauge the openness of the entrance and form the bottleneck of the inhibitor-unbinding pathway. Two special water molecules at the pocket entrance appear to play important roles in inhibitor recognition of binding and unbinding. These water molecules form water bridges between the polar groups of the inhibitor and the residues around the entrance, and between the polar groups of the inhibitor themselves. The water bridged interactions not only induce the inhibitor to adopt an energetically favorable conformation so the inhibitor can pass through the pocket entrance, but also stabilize the binding of the inhibitor in the pocket to prevent the inhibitor's dissociation. The complementary steered molecular dynamics and conventional molecular dynamics simulation results strongly support the hypothesis that NNRTIs inhibit HIV-1 RT polymerization activity by enlarging the DNA-binding cleft and restricting the flexibility and mobility of the p66 thumb subdomain that are believed to be essential during DNA translocation and polymerization. PMID- 12770868 TI - Molecular dynamics simulations of peptides and proteins with amplified collective motions. AB - We present a novel method that uses the collective modes obtained with a coarse grained model/anisotropic network model to guide the atomic-level simulations. Based on this model, local collective modes can be calculated according to a single configuration in the conformational space of the protein. In the molecular dynamics simulations, the motions along the slowest few modes are coupled to a higher temperature by the weak coupling method to amplify the collective motions. This amplified-collective-motion (ACM) method is applied to two test systems. One is an S-peptide analog. We realized the refolding of the denatured peptide in eight simulations out of 10 using the method. The other system is bacteriophage T4 lysozyme. Much more extensive domain motions between the N-terminal and C terminal domain of T4 lysozyme are observed in the ACM simulation compared to a conventional simulation. The ACM method allows for extensive sampling in conformational space while still restricting the sampled configurations within low energy areas. The method can be applied in both explicit and implicit solvent simulations, and may be further applied to important biological problems, such as long timescale functional motions, protein folding/unfolding, and structure prediction. PMID- 12770869 TI - Dielectric self-energy in Poisson-Boltzmann and Poisson-Nernst-Planck models of ion channels. AB - We demonstrated previously that the two continuum theories widely used in modeling biological ion channels give unreliable results when the radius of the conduit is less than two Debye lengths. The reason for this failure is the neglect of surface charges on the protein wall induced by permeating ions. Here we attempt to improve the accuracy of the Poisson-Boltzmann and Poisson-Nernst Planck theories, when applied to channel-like environments, by including a specific dielectric self-energy term to overcome spurious shielding effects inherent in these theories. By comparing results with Brownian dynamics simulations, we show that the inclusion of an additional term in the equations yields significant qualitative improvements. The modified theories perform well in very wide and very narrow channels, but are less successful at intermediate sizes. The situation is worse in multi-ion channels because of the inability of the continuum theories to handle the ion-to-ion interactions correctly. Thus, further work is required if these continuum theories are to be reliably salvaged for quantitative studies of biological ion channels in all situations. PMID- 12770870 TI - Azimuthal frustration and bundling in columnar DNA aggregates. AB - The interaction between two stiff parallel DNA molecules is discussed using linear Debye-Huckel screening theory with and without inclusion of the dielectric discontinuity at the DNA surface, taking into account the helical symmetry of DNA. The pair potential furthermore includes the amount and distribution of counterions adsorbed on the DNA surface. The interaction does not only depend on the interaxial separation of two DNA molecules, but also on their azimuthal orientation. The optimal mutual azimuthal angle is a function of the DNA-DNA interaxial separation, which leads to azimuthal frustrations in an aggregate. On the basis of the pair potential, the positional and orientational order in columnar B-DNA assemblies in solution is investigated. Phase diagrams are calculated using lattice sums supplemented with the entropic contributions of the counterions in solution. A variety of positionally and azimuthally ordered phases and bundling transitions is predicted, which strongly depend on the counterion adsorption patterns. PMID- 12770872 TI - Molecular dynamics simulations of lipid bilayers: major artifacts due to truncating electrostatic interactions. AB - We study the influence of truncating the electrostatic interactions in a fully hydrated pure dipalmitoylphosphatidylcholine (DPPC) bilayer through 20 ns molecular dynamics simulations. The computations in which the electrostatic interactions were truncated are compared to similar simulations using the particle-mesh Ewald (PME) technique. All examined truncation distances (1.8-2.5 nm) lead to major effects on the bilayer properties, such as enhanced order of acyl chains together with decreased areas per lipid. The results obtained using PME, on the other hand, are consistent with experiments. These artifacts are interpreted in terms of radial distribution functions g(r) of molecules and molecular groups in the bilayer plane. Pronounced maxima or minima in g(r) appear exactly at the cutoff distance indicating that the truncation gives rise to artificial ordering between the polar phosphatidyl and choline groups of the DPPC molecules. In systems described using PME, such artificial ordering is not present. PMID- 12770871 TI - Discrete models of autocrine cell communication in epithelial layers. AB - Pattern formation in epithelial layers heavily relies on cell communication by secreted ligands. Whereas the experimentally observed signaling patterns can be visualized at single-cell resolution, a biophysical framework for their interpretation is currently lacking. To this end, we develop a family of discrete models of cell communication in epithelial layers. The models are based on the introduction of cell-to-cell coupling coefficients that characterize the spatial range of intercellular signaling by diffusing ligands. We derive the coupling coefficients as functions of geometric, cellular, and molecular parameters of the ligand transport problem. Using these coupling coefficients, we analyze a nonlinear model of positive feedback between ligand release and binding. In particular, we study criteria of existence of the patterns consisting of clusters of a few signaling cells, as well as the onset of signal propagation. We use our model to interpret recent experimental studies of the EGFR/Rhomboid/Spitz module in Drosophila development. PMID- 12770873 TI - The role of the dielectric barrier in narrow biological channels: a novel composite approach to modeling single-channel currents. AB - A composite continuum theory for calculating ion current through a protein channel of known structure is proposed, which incorporates information about the channel dynamics. The approach is utilized to predict current through the Gramicidin A ion channel, a narrow pore in which the applicability of conventional continuum theories is questionable. The proposed approach utilizes a modified version of Poisson-Nernst-Planck (PNP) theory, termed Potential-of-Mean Force-Poisson-Nernst-Planck theory (PMFPNP), to compute ion currents. As in standard PNP, ion permeation is modeled as a continuum drift-diffusion process in a self-consistent electrostatic potential. In PMFPNP, however, information about the dynamic relaxation of the protein and the surrounding medium is incorporated into the model of ion permeation by including the free energy of inserting a single ion into the channel, i.e., the potential of mean force along the permeation pathway. In this way the dynamic flexibility of the channel environment is approximately accounted for. The PMF profile of the ion along the Gramicidin A channel is obtained by combining an equilibrium molecular dynamics (MD) simulation that samples dynamic protein configurations when an ion resides at a particular location in the channel with a continuum electrostatics calculation of the free energy. The diffusion coefficient of a potassium ion within the channel is also calculated using the MD trajectory. Therefore, except for a reasonable choice of dielectric constants, no direct fitting parameters enter into this model. The results of our study reveal that the channel response to the permeating ion produces significant electrostatic stabilization of the ion inside the channel. The dielectric self-energy of the ion remains essentially unchanged in the course of the MD simulation, indicating that no substantial changes in the protein geometry occur as the ion passes through it. Also, the model accounts for the experimentally observed saturation of ion current with increase of the electrolyte concentration, in contrast to the predictions of standard PNP theory. PMID- 12770874 TI - Two components of voltage-dependent inactivation in Ca(v)1.2 channels revealed by its gating currents. AB - Voltage-dependent inactivation (VDI) was studied through its effects on the voltage sensor in Ca(v)1.2 channels expressed in tsA 201 cells. Two kinetically distinct phases of VDI in onset and recovery suggest the presence of dual VDI processes. Upon increasing duration of conditioning depolarizations, the half distribution potential (V(1/2)) of intramembranous mobile charge was negatively shifted as a sum of two exponential terms, with time constants 0.5 s and 4 s, and relative amplitudes near 50% each. This kinetics behavior was consistent with that of increment of maximal charge related to inactivation (Qn). Recovery from inactivation was also accompanied by a reduction of Qn that varied with recovery time as a sum of two exponentials. The amplitudes of corresponding exponential terms were strongly correlated in onset and recovery, indicating that channels recover rapidly from fast VDI and slowly from slow VDI. Similar to charge "immobilization," the charge moved in the repolarization (OFF) transient became slower during onset of fast VDI. Slow VDI had, instead, hallmarks of interconversion of charge. Confirming the mechanistic duality, fast VDI virtually disappeared when Li(+) carried the current. A nine-state model with parallel fast and slow inactivation pathways from the open state reproduces most of the observations. PMID- 12770875 TI - Pore- and state-dependent cadmium block of I(Ks) channels formed with MinK-55C and wild-type KCNQ1 subunits. AB - Human MinK and KCNQ1 subunits assemble to form I(Ks) channels. When MinK position 55 is mutated to cysteine (MinK-55C), I(Ks) channels can be blocked by external cadmium (Cd(2+)). We have supported a pore-associated location for MinK-55C because Cd(2+) block is sensitive to voltage, permeant ions on the opposite side of the membrane (trans-ions), and external tetraethylammonium (TEA), an I(Ks) pore-blocker. Two recent reports argue that MinK-55C is distant from the pore: one finds TEA does not affect Cd(2+) block if channels are formed with a KCNQ1 mutant (K318I, V319Y) that increases TEA affinity; the second proposes that Cd(2+) binds between MinK-55C and a cysteine in KCNQ1 that is posited to lie toward the channel periphery. Here, these discrepancies are considered. First, Cd(2+) block of MinK-55C channels formed with wild-type KCNQ1 is shown to depend not only on voltage and trans-ions but state (showing decreased on-rate with increased open time and blocker trapping on channel closure). Conversely, MinK 55C channels with K318I, V319Y KCNQ1 are found to demonstrate Cd(2+) block that is independent of voltage, trans-ions and state (and to have a lower unitary conductance): thus, the KCNQ1 mutations alter the process under study, yielding Cd(2+) inhibition that is pore-independent and, perforce, TEA-insensitive. Second, MinK-55C channels are found to remain sensitive to Cd(2+) despite mutation of any single native cysteine in KCNQ1 or all nine simultaneously; this suggests no KCNQ1 cysteine binds Cd(2+) and can serve to localize MinK-55C. Despite many concerns that are enumerated, we remain obliged to conclude that Cd(2+) enters and leaves the pore to reach MinK-55C, placing that residue in or near the pore. PMID- 12770876 TI - Investigating the conformational states of the rabbit Na+/glucose cotransporter. AB - The Na(+) and voltage-dependence of transient rabbit Na(+)/glucose cotransporter (rSGLT1) kinetics was studied with the two-electrode voltage-clamp technique and Xenopus laevis oocytes. Using step changes in membrane potential, in the absence of glucose but with 100 or 10 mM Na(+), transient currents were measured corresponding to binding/debinding of Na(+) and conformational changes of the protein. Previously, only a single time constant has been published for rSGLT1. We, however, observed three decay components; a fast (tau(f), 0.5-1 ms) voltage- and Na(+)-independent decay, and medium (tau(m), 0.5-4 ms) and slow (tau(s), 8-50 ms) voltage- and Na(+)-dependent decays. Transient currents were simulated and fit using a four-state model to obtain kinetic parameters for the system. The four-state model was able to reconstitute an assortment of experimental data. PMID- 12770877 TI - A physical model of potassium channel activation: from energy landscape to gating kinetics. AB - We have developed a method for rapidly computing gating currents from a multiparticle ion channel model. Our approach is appropriate for energy landscapes that can be characterized by a network of well-defined activation pathways with barriers. To illustrate, we represented the gating apparatus of a channel subunit by an interacting pair of charged gating particles. Each particle underwent spatial diffusion along a bistable potential of mean force, with electrostatic forces coupling the two trajectories. After a step in membrane potential, relaxation of the smaller barrier charge led to a time-dependent reduction in the activation barrier of the principal gate charge. The resulting gating current exhibited a rising phase similar to that measured in voltage dependent ion channels. Reduction of the two-dimensional diffusion landscape to a circular Markov model with four states accurately preserved the time course of gating currents on the slow timescale. A composite system containing four subunits leading to a concerted opening transition was used to fit a series of gating currents from the Shaker potassium channel. We end with a critique of the model with regard to current views on potassium channel structure. PMID- 12770878 TI - Contribution of cytosolic cysteine residues to the gating properties of the Kir2.1 inward rectifier. AB - The topological model proposed for the Kir2.1 inward rectifier predicts that seven of the channel 13 cysteine residues are distributed along the N- and C terminus regions, with some of the residues comprised within highly conserved domains involved in channel gating. To determine if cytosolic cysteine residues contribute to the gating properties of Kir2.1, each of the N- and C-terminus cysteines was mutated into either a polar (S, D, N), an aliphatic (A,V, L), or an aromatic (W) residue. Our patch-clamp measurements show that with the exception of C76 and C311, the mutation of individual cytosolic cysteine to serine (S) did not significantly affect the single-channel conductance nor the channel open probability. However, mutating C76 to a charged or polar residue resulted either in an absence of channel activity or a decrease in open probability. In turn, the mutations C311S (polar), C311R (charged), and to a lesser degree C311A (aliphatic) led to an increase of the channel mean closed time due to the appearance of long closed time intervals (T(c) >or= 500 ms) and to a reduction of the reactivation by ATP of rundown Kir2.1 channels. These changes could be correlated with a weakening of the interaction between Kir2.1 and PIP(2), with C311R and C311S being more potent at modulating the Kir2.1-PIP(2) interaction than C311A. The present work supports, therefore, molecular models whereby the gating properties of Kir2.1 depend on the presence of nonpolar or neutral residues at positions 76 and 311, with C311 modulating the interaction between Kir2.1 and PIP(2). PMID- 12770879 TI - The effect of lipid demixing on the electrostatic interaction of planar membranes across a salt solution. AB - We study the effect of lipid demixing on the electrostatic interaction of two oppositely-charged membranes in solution, modeled here as an incompressible two dimensional fluid mixture of neutral and charged mobile lipids. We calculate, within linear and nonlinear Poisson-Boltzmann theory, the membrane separation at which the net electrostatic force between the membranes vanishes, for a variety of different system parameters. According to Parsegian and Gingell, contact between oppositely-charged surfaces in an electrolyte is possible only if the two surfaces have exactly the same charge density (sigma(1) = -sigma(2)). If this condition is not fulfilled, the surfaces can repel each other, even though they are oppositely charged. In our model of a membrane, the lipidic charge distribution on the membrane surface is not homogeneous and frozen, but the lipids are allowed to freely move within the plane of the membrane. We show that lipid demixing allows contact between membranes even if there is a certain charge mismatch, /sigma(1)/ not equal /sigma(2)/, and that in certain limiting cases, contact is always possible, regardless of the value of sigma(1)/sigma(2) (if sigma(1)/sigma(2) < 0). We furthermore find that of the two interacting membranes, only one membrane shows a major rearrangement of lipids, whereas the other remains in exactly the same state it has in isolation and that, at zero disjoining pressure, the electrostatic mean-field potential between the membranes follows a Gouy-Chapman potential from the more strongly charged membrane up to the point of the other, more weakly charged membrane. PMID- 12770880 TI - Molecular dynamics simulation of a dipalmitoylphosphatidylcholine bilayer with NaCl. AB - Molecular dynamics simulations are performed on two hydrated dipalmitoylphosphatidylcholine bilayer systems: one with pure water and one with added NaCl. Due to the rugged nature of the membrane/electrolyte interface, ion binding to the membrane surface is characterized by the loss of ion hydration. Using this structural characterization, binding of Na(+) and Cl(-) ions to the membrane is observed, although the binding of Cl(-) is seen to be slightly weaker than that of Na(+). Dehydration is seen to occur to a different extent for each type of ion. In addition, the excess binding of Na(+) gives rise to a net positive surface charge density just outside the bilayer. The positive density produces a positive electrostatic potential in this region, whereas the system without salt shows an electrostatic potential of zero. PMID- 12770882 TI - The effect of fructan on membrane lipid organization and dynamics in the dry state. AB - Fructans are a group of fructose-based oligo- and polysaccharides, which appear to be involved in membrane preservation during dehydration by interacting with the membrane lipids. To get further understanding of the protective mechanism, the consequences of the fructan-membrane lipid interaction for the molecular organization and dynamics in the dry state were studied. POPC and DMPC were investigated in the dry state by (2)H, (31)P NMR, and Fourier transform infrared spectroscopy using two types of fructan and dextran. The order-disorder transition temperature of dry POPC was reduced by 70 degrees C in the presence of fructan. Fructan increased the mobility of the acyl chains, but immobilized the lipid headgroup region. Most likely, fructans insert between the headgroups of lipids, thereby spacing the acyl chains. This results in a much lower phase transition temperature. The headgroup is immobilized by the interaction with fructan. The location of the interaction with the lipid headgroup is different for the inulin-type fructan compared to the levan-type fructan, since inulin shows interaction with the lipid phosphate group, whereas levan does not. Dextran did not influence the phase transition temperature of dry POPC showing that reduction of this temperature is not a general property of polysaccharides. PMID- 12770881 TI - Evidence for membrane thinning effect as the mechanism for peptide-induced pore formation. AB - Antimicrobial peptides have two binding states in a lipid bilayer, a surface state S and a pore-forming state I. The transition from the S state to the I state has a sigmoidal peptide-concentration dependence indicating cooperativity in the peptide-membrane interactions. In a previous paper, we reported the transition of alamethicin measured in three bilayer conditions. The data were explained by a free energy that took into account the membrane thinning effect induced by the peptides. In this paper, the full implications of the free energy were tested by including another type of peptide, melittin, that forms toroidal pores, instead of barrel-stave pores as in the case of alamethicin. The S-to-I transitions were measured by oriented circular dichroism. The membrane thinning effect was measured by x-ray diffraction. All data were in good agreement with the theory, indicating that the membrane thinning effect is a plausible mechanism for the peptide-induced pore formations. PMID- 12770883 TI - Collective chain dynamics in lipid bilayers by inelastic x-ray scattering. AB - We investigated the application of inelastic x-ray scattering (IXS) to lipid bilayers. This technique directly measures the dynamic structure factor S(q,omega) which is the space-time Fourier transform of the electron density correlation function of the measured system. For a multiatomic system, the analysis of S(q,omega) is usually complicated. But for multiple bilayers of lipid, S(q,omega) is dominated by chain-chain correlations within individual bilayers. Thus IXS provides a unique probe for the collective dynamics of lipid chains in a bilayer that cannot be obtained by any other method. IXS of dimyristoyl phosphatidylcholine and dimyristoyl phosphatidylcholine + cholesterol at two different concentrations were measured. S(q,omega) was analyzed by three mode hydrodynamic equations, including a thermal diffusive mode and two propagating acoustic modes. We obtained the dispersion curves for the phonons that represent the collective in-plane excitations of lipid chains. The effect of cholesterol on chain dynamics was detected. Our analysis shows the importance of having a high instrument resolution as well as the requirement of sufficient signal-to-noise ratio to obtain meaningful results from such an IXS experiment. The requirement on signal-to-noise also applies to molecular dynamics simulations. PMID- 12770884 TI - Time-resolved fluorescence and fourier transform infrared spectroscopic investigations of lateral packing defects and superlattice domains in compositionally uniform cholesterol/phosphatidylcholine bilayers. AB - Time-resolved fluorescence and Fourier transform infrared spectroscopies were used to investigate the lateral organization of lipids in compositionally uniform and fully equilibrated 1-palmitoyl-2-oleoyl-phosphatidylcholine/cholesterol (POPC/CHOL) liposomes prepared by a recently devised low-temperature trapping method. Independent fluorescence decay lifetime and rotational dynamics parameters of diphenylhexatriene (DPH) chain-labeled phosphatidylcholine (DPH-PC) in these liposomes were recovered from the time-resolved fluorescence measurements as a function of cholesterol molar fraction (X(CHOL)) at 23 degrees C. The results indicate significantly greater lifetime heterogeneity, shorter average lifetime, rotational correlation time, and lower order parameter of the DPH moiety at X(CHOL) approximately 0.40 and 0.50 as compared to the adjacent cholesterol concentrations. Less prominent changes were also detected at, for example, X(CHOL) approximately 0.20 and 0.33. These X(CHOL)'s coincide with the "critical" X(CHOL)'s predicted by the previously proposed superlattice (SL) model, thus indicating that POPC and cholesterol molecules tend to form SL domains where the components tend to be regularly distributed. The data also support another prediction of the SL model, namely that lateral packing defects coexist with the ordered SL domains. It appears that unfavorable interaction of the DPH-moiety of DPH-PC with cholesterol results in a preferential partition of DPH-PC to the defect regions. Fourier transform infrared analysis of the native lipid O=P=O, C=O, and C-H vibrational bands of POPC/CHOL liposomes in the absence of DPH-PC revealed an increase in the conformational order of the acyl chains and a decrease in the conformational order (or increased hydration) of the interfacial and headgroup regions at or close to the predicted critical X(CHOL)'s. This provides additional but probe-independent evidence for SL domain formations in the POPC/CHOL bilayers. We propose that the defect regions surrounding the putative SL domains could play an important role in modulating the activity of various membrane-associated enzymes, e.g., those regulating the lipid compositions of cell membranes. PMID- 12770885 TI - Liquid-crystalline collapse of pulmonary surfactant monolayers. AB - During exhalation, the surfactant film of lipids and proteins that coats the alveoli in the lung is compressed to high surface pressures, and can remain metastable for prolonged periods at pressures approaching 70 mN/m. Monolayers of calf lung surfactant extract (CLSE), however, collapse in vitro, during an initial compression at approximately 45 mN/m. To gain information on the source of this discrepancy, we investigated how monolayers of CLSE collapse from the interface. Observations with fluorescence, Brewster angle, and light scattering microscopies show that monolayers containing CLSE, CLSE-cholesterol (20%), or binary mixtures of dipalmitoyl phosphatidylcholine(DPPC)-dihydrocholesterol all form bilayer disks that reside above the monolayer. Upon compression and expansion, lipids flow continuously from the monolayer into the disks, and vice versa. In several respects, the mode of collapse resembles the behavior of other amphiphiles that form smectic liquid-crystal phases. These findings suggest that components of surfactent films must collapse collectively rather than being squeezed out individually. PMID- 12770886 TI - Cycling cross-bridges increase myocardial stiffness at submaximal levels of Ca2+ activation. AB - Permeabilized multicellular preparations of canine myocardium were subjected to controlled length changes to investigate the extent to which cross-bridges augment passive stiffness components in myocardium at low levels of Ca(2+) activation. When the preparations were immersed in pCa 9.0 solution (negligible free [Ca(2+)]) they behaved as simple elastic systems (i.e., tension increased proportionately with length). In contrast, when the muscles were stretched in Ca(2+) activating solutions, tension rose much more rapidly during the initial phase of the movement than thereafter. Several lines of evidence suggest that the nonlinear response represents the displacement of populations of cycling cross bridges that are perturbed by interfilamentary movement and take some time to recover. 1), The stiffness of the initial phase increased proportionately with the level of Ca(2+) activation. 2), The magnitude of the short-range response increased with stretch velocity. 3), The initial response was reversibly reduced by 5-mM 2,3-butanedione monoxime, a known cross-bridge inhibitor. The initial stiffness of the passive elastic (pCa 9.0) response was equivalent to the Ca(2+) dependent component at 2% (pCa approximately 6.2) of the maximal (pCa 4.5) level. These results suggest that cross-bridges may significantly affect diastolic chamber stiffness. PMID- 12770888 TI - The flexibility of DNA double crossover molecules. AB - Double crossover molecules are DNA structures containing two Holliday junctions connected by two double helical arms. There are several types of double crossover molecules, differentiated by the relative orientations of their helix axes, parallel or antiparallel, and by the number of double helical half-turns (even or odd) between the two crossovers. They are found as intermediates in meiosis and they have been used extensively in structural DNA nanotechnology for the construction of one-dimensional and two-dimensional arrays and in a DNA nanomechanical device. Whereas the parallel double helical molecules are usually not well behaved, we have focused on the antiparallel molecules; antiparallel molecules with an even number of half-turns between crossovers (termed DAE molecules) produce a reporter strand when ligated, facilitating their characterization in a ligation cyclization assay. Hence, we have estimated the flexibility of antiparallel DNA double crossover molecules by means of ligation closure experiments. We are able to show that these molecules are approximately twice as rigid as linear duplex DNA. PMID- 12770887 TI - Cardiac-type EC-coupling in dysgenic myotubes restored with Ca2+ channel subunit isoforms alpha1C and alpha1D does not correlate with current density. AB - Ca(2+)-induced Ca(2+)-release (CICR)-the mechanism of cardiac excitation contraction (EC) coupling-also contributes to skeletal muscle contraction; however, its properties are still poorly understood. CICR in skeletal muscle can be induced independently of direct, calcium-independent activation of sarcoplasmic reticulum Ca(2+) release, by reconstituting dysgenic myotubes with the cardiac Ca(2+) channel alpha(1C) (Ca(V)1.2) subunit. Ca(2+) influx through alpha(1C) provides the trigger for opening the sarcoplasmic reticulum Ca(2+) release channels. Here we show that also the Ca(2+) channel alpha(1D) isoform (Ca(V)1.3) can restore cardiac-type EC-coupling. GFP-alpha(1D) expressed in dysgenic myotubes is correctly targeted into the triad junctions and generates action potential-induced Ca(2+) transients with the same efficiency as GFP alpha(1C) despite threefold smaller Ca(2+) currents. In contrast, GFP-alpha(1A), which generates large currents but is not targeted into triads, rarely restores action potential-induced Ca(2+) transients. Thus, cardiac-type EC-coupling in skeletal myotubes depends primarily on the correct targeting of the voltage-gated Ca(2+) channels and less on their current size. Combined patch-clamp/fluo-4 Ca(2+) recordings revealed that the induction of Ca(2+) transients and their maximal amplitudes are independent of the different current densities of GFP alpha(1C) and GFP-alpha(1D). These properties of cardiac-type EC-coupling in dysgenic myotubes are consistent with a CICR mechanism under the control of local Ca(2+) gradients in the triad junctions. PMID- 12770889 TI - Stability of the I-motif structure is related to the interactions between phosphodiester backbones. AB - The i-motif DNA tetrameric structure is formed of two parallel duplexes intercalated in a head-to-tail orientation, and held together by hemiprotonated cytosine pairs. The four phosphodiester backbones forming the structure define two narrow and wide grooves. The short interphosphate distances across the narrow groove induce a strong repulsion which should destabilize the tetramer. To investigate this point, molecular dynamics simulations were run on the [d(C2)]4 and [d(C4)]4 tetramers in 3'E and 5'E topologies, for which the interaction of the phosphodiester backbones through the narrow groove is different. The analysis of the simulations, using the Molecular Mechanics Generalized Born Solvation Area and Molecular Mechanics Poisson-Boltzmann Solvation Area approaches, shows that it is the van der Waals energy contribution which displays the largest relative difference between the two topologies. The comparison of the solvent-accessible area of each topology reveals that the sugar-sugar interactions account for the greater stability of the 3'E topology. This stresses the importance of the sugar sugar contacts across the narrow groove which, enforcing the optimal backbone twisting, are essential to the base stacking and the i-motif stability. Tighter interactions between the sugars are observed in the case of N-type sugar puckers. PMID- 12770890 TI - Late events in the photocycle of bacteriorhodopsin mutant L93A. AB - In the photocycle of bacteriorhodopsin (bR) from Halobacterium salinarum mutant L93A, the O-intermediate accumulates and the cycling time is increased approximately 200 times. Nevertheless, under continuous illumination, the protein pumps protons at near wild-type rates. We excited the mutant L93A in purple membrane with single or triple laser flashes and quasicontinuous illumination, (i.e., light for a few seconds) and recorded proton release and uptake, electric signals, and absorbance changes. We found long-living, correlated, kinetic components in all three measurements, which-with exception of the absorbance changes-had not been seen in earlier investigations. At room temperature, the O intermediate decays to bR in two transitions with rate constants of 350 and 1800 ms. Proton uptake from the cytoplasmic surface continues with similar kinetics until the bR state is reestablished. An analysis of the data from quasicontinuous illumination and multiple flash excitation led to the conclusion that acceleration of the photocycle in continuous light is due to excitation of the N component in the fast N<-->O equilibrium, which is established at the beginning of the severe cycle slowdown. This conclusion was confirmed by an action spectrum. PMID- 12770891 TI - Photoexcitation of the O-intermediate in bacteriorhodopsin mutant L93A. AB - During the extended lifetime of the O-state in bacteriorhodopsin (bR) mutant L93A, two substates have been distinguished. The first O-intermediate (OI) is in rapid equilibrium with N and apparently still has a 13-cis chromophore. OI undergoes a photoreaction with a small absorbance change, positive charge transport in the pumping direction, and proton release and uptake. None of these effects was detected after photoexcitation of the late O (OII). The most likely interpretation of the effects seen is an accelerated return of the molecule from the OI- to the bR-state. However, with a lifetime approximately 140 ms, the reaction cannot account for the observed high pumping efficiency of the mutant under continuous illumination. We suggest that OII corresponds to the O intermediate with a twisted all-trans chromophore seen in the photocycle of wild type bR, where the 13-cis OI-intermediate under the usual conditions does not accumulate in easily detectable amounts and, therefore, has generally been overlooked. Both the OI- and OII-decays are apparently strongly inhibited in the mutant. PMID- 12770892 TI - Electric-field dependent decays of two spectroscopically different M-states of photosensory rhodopsin II from Natronobacterium pharaonis. AB - Sensory rhodopsin II (NpSRII) from Natronobacterium pharaonis was studied by resonance Raman (RR) spectroscopic techniques. Using gated 413-nm excitation, time-resolved RR measurements of the solubilized photoreceptor were carried out to probe the photocycle intermediates that are formed in the submillisecond time range. For the first time, two M-like intermediates were identified on the basis of their C=C stretching bands at 1568 and 1583 cm(-1), corresponding to the early M(L)(400) state with a lifetime of 30 micro s and the subsequent M(1)(400) state with a lifetime of 2 ms, respectively. The unusually high C=C stretching frequency of M(1)(400) has been attributed to an unprotonated retinal Schiff base in a largely hydrophobic environment, implying that the M(L)(400) --> M(1)(400) transition is associated with protein structural changes in the vicinity of the chromophore binding pocket. Time-resolved surface enhanced resonance Raman experiments of NpSRII electrostatically bound onto a rotating Ag electrode reveal that the photoreceptor runs through the photocycle also in the immobilized state. Surface enhanced resonance Raman spectra are very similar to the RR spectra of the solubilized protein, ruling out adsorption-induced structural changes in the retinal binding pocket. The photocycle kinetics, however, is sensitively affected by the electrode potential such that at 0.0 V (versus Ag/AgCl) the decay times of M(L)(400) and M(1)(400) are drastically slowed down. Upon decreasing the potential to -0.4 V, that corresponds to a decrease of the interfacial potential drop and thus of the electric field strength at the protein binding site, the photocycle kinetics becomes similar to that of NpSRII in solution. The electric field dependence of the protein structural changes associated with the M-state transitions, which in the present spectroscopic work is revealed on a molecular level, appears to be related to the electric-field control of bacteriorhodopsin's photocycle, which has been shown to be of functional relevance. PMID- 12770893 TI - Nanosecond temperature jump relaxation dynamics of cyclic beta-hairpin peptides. AB - The thermal unfolding of a series of 6-, 10-, and 14-mer cyclic beta-hairpin peptides was studied to gain insight into the mechanism of formation of this important secondary structure. The thermodynamics of the transition were characterized using temperature dependent Fourier transform infrared spectroscopy. Thermodynamic data were analyzed using a two-state model which indicates increasing cooperativity along the series. The relaxation kinetics of the peptides in response to a laser induced temperature jump were probed using time-resolved infrared spectroscopy. Single exponential relaxation kinetics were observed and fit with a two-state model. The folding rate determined for these cyclic peptides is accelerated by some two orders of magnitude over the rate of a linear peptide that forms a beta-hairpin. This observation supports the argument that the rate limiting step in the linear system is either stabilization of compact collapsed structures or rearrangement of collapsed structures over a barrier to achieve the native interstrand registry. Small activation energies for folding of these peptides obtained from an Arrhenius analysis of the rates imply a primarily entropic barrier, hence an organized transition state having specific stabilizing interactions. PMID- 12770894 TI - Molecular dynamics simulations of lignin peroxidase in solution. AB - The dynamical and structural properties of lignin peroxidase and its Trp171Ala mutant have been investigated in aqueous solution using molecular dynamics (MD) simulations. In both cases, the enzyme retained its overall backbone structure and all its noncovalent interactions in the course of the MD simulations. Very interestingly, the analysis of the MD trajectories showed the presence of large fluctuations in correspondence of the residues forming the heme access channel; these movements enlarge the opening and facilitate the access of substrates to the enzyme active site. Moreover, steered molecular dynamics docking simulations have shown that lignin peroxidase natural substrate (veratryl alcohol) can easily approach the heme edge through the access channel. PMID- 12770895 TI - Structural studies of MS2 bacteriophage virus particle disassembly by nuclear magnetic resonance relaxation measurements. AB - In this article we studied, by nuclear magnetic resonance relaxation measurements, the disassembly of a virus particle-the MS2 bacteriophage. MS2 is one of the single-stranded RNA bacteriophages that infect Escherichia coli. At pH 4.5, the phage turns to a metastable state, as is indicated by an increase in the observed nuclear magnetic resonance signal intensity upon decreasing the pH from 7.0 to 4.5. Steady-state fluorescence and circular dichroism spectra at pH 4.5 show that the difference in conformation and secondary structure is not pronounced if compared with the phage at pH 7.0. At pH 4.5, two-dimensional (15)N (1)H heteronuclear multiple quantum coherence (HMQC) spectrum shows approximately 40 crosspeaks, corresponding to the most mobile residues of MS2 coat protein at pH 4.5. The (15)N linewidth is approximately 30 Hz, which is consistent with an intermediate with a rotational relaxation time of 100 ns. The average spin lattice relaxation time (T(1)) of the mobile residues was measured at different temperatures, clearly distinguishing between the dimer and the equilibrium intermediate. The results show, for the first time, the presence of intermediates in the process of dissociation of the MS2 bacteriophage. PMID- 12770896 TI - Refolding of a high molecular weight protein: salt effect on collapse. AB - Small-angle neutron scattering experiments were performed on dilute solutions of a high molecular weight protein (fibronectin, M = 580 kg/mol) in four cases: native conditions; unfolded state obtained by a denaturing agent (urea); and two badly refolded (or collapsed) states obtained by progressive elimination of the denaturing agent in salt-containing or salt-free solutions. Our main result is concerned by the conformation of the protein as the attempt for refolding is driven with or without salt. In salt-containing solution, we observe unambiguously that the protein chain collapses at large length scales but still obeys to a Gaussian statistics at short length scales. In other words, the globule embodies a large quantity of solvent compared to the compact situation. In salt-free solutions, the badly refolded protein is not globular but displays both a coil-like and an open conformation at large length scales and a local high density area. This behavior is discussed with respect to the scaling theories for polymers and polyampholytes. PMID- 12770897 TI - 15N-1H Residual dipolar coupling analysis of native and alkaline-K79A Saccharomyces cerevisiae cytochrome c. AB - Residual dipolar couplings (RDCs) and pseudocontact shifts are experimentally accessible properties in nuclear magnetic resonance that are related to structural parameters and to the magnetic susceptibility anisotropy. We have determined RDCs due to field-induced orientation of oxidized-K79A and reduced cytochrome c at pH 7.0 and oxidized-K79A cytochrome c at pH 11.1 through measurements of amide (15)N-(1)H (1)J couplings at 800 and 500 MHz. The pH 7.0 RDCs for Fe(III)- and Fe(II)-cytochrome c together with available nuclear Overhauser effects were used to recalculate solution structures that were consistent with both sets of constraints. Molecular magnetic susceptibility anisotropy values were calculated for both redox states of the protein. By subtracting the residual dipolar couplings (RDCs) of the reduced form from those of the oxidized form measured at the same magnetic field (800 MHz), we found the RDC contribution of the paramagnetic metal ion in the oxidized protein. The magnetic susceptibility anisotropy, which was calculated from the structure, was found to be the same as that of the paramagnetic metal ion obtained independently from pseudocontact shifts, thereby indicating that the elements of secondary structure either are rigid or display the same mobility in both oxidation states. The residual dipolar coupling values of the alkaline-K79A form are small with respect to those of oxidized native cytochrome, whereas the pseudocontact shifts are essentially of the same magnitude, indicating local mobility. Importantly, this is the first time that mobility has been found through comparison of RDCs with pseudocontact shifts. PMID- 12770898 TI - A measure of conformational entropy change during thermal protein unfolding using neutron spectroscopy. AB - Thermal unfolding of proteins at high temperatures is caused by a strong increase of the entropy change which lowers Gibbs free energy change of the unfolding transition (DeltaG(unf) = DeltaH - TDeltaS). The main contributions to entropy are the conformational entropy of the polypeptide chain itself and ordering of water molecules around hydrophobic side chains of the protein. To elucidate the role of conformational entropy upon thermal unfolding in more detail, conformational dynamics in the time regime of picoseconds was investigated with neutron spectroscopy. Confined internal structural fluctuations were analyzed for alpha-amylase in the folded and the unfolded state as a function of temperature. A strong difference in structural fluctuations between the folded and the unfolded state was observed at 30 degrees C, which increased even more with rising temperatures. A simple analytical model was used to quantify the differences of the conformational space explored by the observed protein dynamics for the folded and unfolded state. Conformational entropy changes, calculated on the basis of the applied model, show a significant increase upon heating. In contrast to indirect estimates, which proposed a temperature independent conformational entropy change, the measurements presented here, demonstrated that the conformational entropy change increases with rising temperature and therefore contributes to thermal unfolding. PMID- 12770899 TI - Direct measurement of equilibrium constants for high-affinity hemoglobins. AB - The biological functions of heme proteins are linked to their rate and affinity constants for ligand binding. Kinetic experiments are commonly used to measure equilibrium constants for traditional hemoglobins comprised of pentacoordinate ligand binding sites and simple bimolecular reaction schemes. However, kinetic methods do not always yield reliable equilibrium constants with more complex hemoglobins for which reaction mechanisms are not clearly understood. Furthermore, even where reaction mechanisms are clearly understood, it is very difficult to directly measure equilibrium constants for oxygen and carbon monoxide binding to high-affinity (K(D) << 1 micro M) hemoglobins. This work presents a method for direct measurement of equilibrium constants for high affinity hemoglobins that utilizes a competition for ligands between the "target" protein and an array of "scavenger" hemoglobins with known affinities. This method is described for oxygen and carbon monoxide binding to two hexacoordinate hemoglobins: rice nonsymbiotic hemoglobin and Synechocystis hemoglobin. Our results demonstrate that although these proteins have different mechanisms for ligand binding, their affinities for oxygen and carbon monoxide are similar. Their large affinity constants for oxygen, 285 and approximately 100 micro M(-1) respectively, indicate that they are not capable of facilitating oxygen transport. PMID- 12770900 TI - Protein self-association in solution: the bovine pancreatic trypsin inhibitor decamer. AB - We have used magnetic relaxation dispersion to study bovine pancreatic trypsin inhibitor (BPTI) self-association as a function of pH, salt type and concentration, and temperature. The magnetic relaxation dispersion method sensitively detects stable oligomers without being affected by other interactions. We find that BPTI decamers form cooperatively under a wide range of solution conditions with no sign of dimers or other small oligomers. Decamer formation is opposed by electrostatic repulsion among numerous cationic residues confined within a narrow channel. Accordingly, the decamer population increases with increasing pH, as cationic residues are deprotonated, and with increasing salt concentration. The salt effect cannot be described in terms of Debye screening, but involves the ion-specific sequestering of anions within the narrow channel. The lifetime of the BPTI decamer is 101 +/- 4 min at 27 degrees C. We propose that the BPTI decamer, with a heparin chain threading the decamer channel, plays a functional role in the mast cell. We also detect a higher oligomer that appears to be a subcritical nucleation cluster of 3-5 decamers. We argue that monomeric crystals form at high pH despite a high decamer population in solution, because the ion pairs that provide the critical decamer-decamer contacts are disrupted at high pH. PMID- 12770901 TI - Mobility of taxol in microtubule bundles. AB - Mobility of taxol inside microtubules was investigated using fluorescence recovery after photobleaching on flow-aligned bundles. Bundles were made of microtubules with either GMPCPP or GTP at the exchangeable site on the tubulin dimer. Recovery times were sensitive to bundle thickness and packing, indicating that taxol molecules are able to move laterally through the bundle. The density of open binding sites along a microtubule was varied by controlling the concentration of taxol in solution for GMPCPP samples. With >63% sites occupied, recovery times were independent of taxol concentration and, therefore, inversely proportional to the microscopic dissociation rate, k(off). It was found that 10k(off)(GMPCPP) approximately equal k(off)(GTP), consistent with, but not fully accounting for, the difference in equilibrium constants for taxol on GMPCPP and GTP microtubules. With <63% sites occupied, recovery times decreased as approximately [Tax](-1/5) for both types of microtubules. We conclude that the diffusion of taxol inside the microtubule bundle is hindered by rebinding events when open sites are within approximately 7 nm of each other. PMID- 12770902 TI - Nonresonant confocal Raman imaging of DNA and protein distribution in apoptotic cells. AB - Nonresonant confocal Raman imaging has been used to map the DNA and the protein distributions in individual single human cells. The images are obtained on an improved homebuilt confocal Raman microscope. After statistical analysis, using singular value decomposition, the Raman images are reconstructed from the spectra covering the fingerprint region. The data are obtained at a step interval of approximately 250 nm and cover a field from 8- to 15- micro m square in size. Dwell times at each pixel are between 0.5 and 2 s, depending on the nature and the state of the cell under investigation. High quality nonresonant Raman images can only be obtained under these conditions using continuous wave high laser powers between 60 and 120 mW. We will present evidence that these laser powers can still safely be used to recover the chemical distributions in fixed cells. The developed Raman imaging method is used to image directly, i.e., without prior labeling, the nucleotide condensation and the protein distribution in the so called nuclear fragments of apoptotic HeLa cells. In the control (nonapoptotic) HeLa cells, we show, for the first time by Raman microspectroscopy, the presence of the RNA in a cell nucleus. PMID- 12770903 TI - Connecting nanoscale images of proteins with their genetic sequences. AB - We present a technique for reconstructing biomolecular structures from scanning force microscope data. The technique works by iteratively refining model molecules by comparison of simulated and experimental images. It can remove instrument artifacts to yield accurate dimensional measurements from tip broadened data. The result of the reconstruction is a model that can be chosen to include the physically significant parameters for the system at hand. We demonstrate this by reconstructing scanning force microscope images of the cartilage proteoglycan aggrecan. By explicitly including the protein backbone in the model, we are able to associate measured three-dimensional structures with sites in the protein primary structure. The distribution of aggrecan core protein lengths that we measure suggests that 48% of aggrecan molecules found in vivo have been partially catabolized at either the E(1480)-(1481)G or E(1667)-(1668)G aggrecanase cleavage site. PMID- 12770904 TI - FRET or no FRET: a quantitative comparison. AB - Fluorescence resonance energy transfer (FRET) is a technique used to measure the interaction between two molecules labeled with two different fluorophores (the donor and the acceptor) by the transfer of energy from the excited donor to the acceptor. In biological applications, this technique has become popular to qualitatively map protein-protein interactions, and in biophysical projects it is used as a quantitative measure for distances between a single donor and acceptor molecule. Numerous approaches can be found in the literature to quantify and map FRET, but the measures they provide are often difficult to interpret. We propose here a quantitative comparison of these methods by using a surface FRET system with controlled amounts of donor and acceptor fluorophores and controlled distances between them. We support the system with a Monte Carlo simulation of FRET, which provides reference values for the FRET efficiency under various experimental conditions. We validate a representative set of FRET efficiencies and indices calculated from the different methods with different experimental settings. Finally, we test their sensitivity and draw conclusions for the preparation of FRET experiments in more complex and less-controlled systems. PMID- 12770905 TI - Isolation of bright aggregate fluctuations in a multipopulation image correlation spectroscopy system using intensity subtraction. AB - Image correlation spectroscopy allows sensitive measurement of the spatial distribution and aggregation state of fluorescent membrane macro molecules. When studying a single population system (i.e., aggregates of similar brightness), an accurate measure can be made of the aggregate number per observation area, but this measurement becomes much more complex in a distributed population system (i.e., bright and faint aggregates). This article describes an alternate solution that involves extraction of the bright aggregate population information. This novel development for image correlation spectroscopy, termed intensity subtraction analysis, uses sequential uniform intensity subtraction from raw confocal images. Sequential intensity subtraction results in loss of faint aggregate fluctuations that are smaller in magnitude than fluctuations due to the brightest aggregates. The resulting image has correlatable fluctuations originating from only the brightest population, permitting quantification of this population's distribution and further cross-correlation measurements. The feasibility of this technique is demonstrated using fluorescent microsphere images and biological samples. The technique is further used to examine the spatial distribution of a plasma-membrane-labeled fluorescent synthetic ganglioside, and to cross-correlate this probe with various membrane markers. The evidence provided demonstrates that bright aggregates of the fluorescent ganglioside are associated with clathrin-coated pits, membrane microvilli, and detergent-resistant membranes. PMID- 12770906 TI - Selective cell targeting with light-absorbing microparticles and nanoparticles. AB - We describe a new method for selective cell targeting based on the use of light absorbing microparticles and nanoparticles that are heated by short laser pulses to create highly localized cell damage. The method is closely related to chromophore-assisted laser inactivation and photodynamic therapy, but is driven solely by light absorption, without the need for photochemical intermediates (particularly singlet oxygen). The mechanism of light-particle interaction was investigated by nanosecond time-resolved microscopy and by thermal modeling. The extent of light-induced damage was investigated by cell lethality, by cell membrane permeability, and by protein inactivation. Strong particle size dependence was found for these interactions. A technique based on light to target endogenous particles is already being exploited to treat pigmented cells in dermatology and ophthalmology. With exogenous particles, phamacokinetics and biodistribution studies are needed before the method can be evaluated against photodynamic therapy for cancer treatment. However, particles are unique, unlike photosensitizers, in that they can remain stable and inert in cells for extended periods. Thus they may be particularly useful for prelabeling cells in engineered tissue before implantation. Subsequent irradiation with laser pulses will allow control of the implanted cells (inactivation or modulation) in a noninvasive manner. PMID- 12770907 TI - Functional analysis of the structural basis of homophilic cadherin adhesion. AB - The structures of many cell surface adhesion proteins comprise multiple tandem repeats of structurally similar domains. In many cases, the functional significance of this architecture is unknown, and there are several cases in which evidence for individual domain involvement in adhesion has been contradictory. In particular, the extracellular region of the adhesion glycoprotein cadherin consists of five tandemly arranged domains. One proposed mechanism postulated that adhesion involves only trans interactions between the outermost domains. However, subsequent investigations have generated several competing models. Here we describe direct measurements of the distance-dependent interaction potentials between cadherin mutants lacking different domains. By quantifying both the absolute distances at which opposed cadherin fragments bind and the quantized changes in the interaction potentials that result from deletions of individual domains, we demonstrate that two domains participate in homophilic cadherin binding. This finding contrasts with the current view that cadherins bind via a single, unique site on the protein surface. The potentials that result from interactions involving multiple domains generate a novel, modular binding mechanism in which opposed cadherin ectodomains can adhere in any of three antiparallel alignments. PMID- 12770908 TI - Anomalous protein diffusion in living cells as seen by fluorescence correlation spectroscopy. AB - We investigate the challenges and limitations that are encountered when studying membrane protein dynamics in vivo by means of fluorescence correlation spectroscopy (FCS). Based on theoretical arguments and computer simulations, we show that, in general, the fluctuating fluorescence has a fractal dimension D(0) >or= 1.5, which is determined by the anomality alpha of the diffusional motion of the labeled particles, i.e., by the growth of their mean square displacement as (Deltax)(2) approximately t(alpha). The fractality enforces an initial power-law behavior of the autocorrelation function and related quantities for small times. Using this information, we show by FCS that Golgi resident membrane proteins move subdiffusively in the endoplasmic reticulum and the Golgi apparatus in vivo. Based on Monte Carlo simulations for FCS on curved surfaces, we can rule out that the observed anomalous diffusion is a result of the complex topology of the membrane. The apparent mobility of particles as determined by FCS, however, is shown to depend crucially on the shape of the membrane and its motion in time. Due to this fact, the hydrodynamic radius of the tracked particles can be easily overestimated by an order of magnitude. PMID- 12770909 TI - Single molecule detection of DNA looping by NgoMIV restriction endonuclease. AB - Single molecule fluorescence resonance energy transfer (FRET) and fluorescence correlation spectroscopy were used to investigate DNA looping by NgoMIV restriction endonuclease. Using a linear double-stranded DNA (dsDNA) molecule labeled with a fluorescence donor molecule, Cy3, and fluorescence acceptor molecule, Cy5, and by varying the concentration of NgoMIV endonuclease from 0 to 3 x 10(-6) M, it was possible to detect and determine diffusion properties of looped DNA/protein complexes. FRET efficiency distributions revealed a subpopulation of complexes with an energy transfer efficiency of 30%, which appeared upon addition of enzyme in the picomolar to nanomolar concentration range (using 10(-11) M dsDNA). The concentration dependence, fluorescence burst size analysis, and fluorescence correlation analysis were all consistent with this subpopulation arising from a sequence specific interaction between an individual enzyme and a DNA molecule. A 30% FRET efficiency corresponds to a distance of approximately 65 A, which correlates well with the distance between the ends of the dsDNA molecule when bound to NgoMIV according to the crystal structure of this complex. Formation of the looped complexes was also evident in measurements of the diffusion times of freely diffusing DNA molecules with and without NgoMIV. At very high protein concentrations compared to the DNA concentration, FRET and fluorescence correlation spectroscopy results revealed the formation of larger DNA/protein complexes. PMID- 12770910 TI - Combined affinity and rate constant distributions of ligand populations from experimental surface binding kinetics and equilibria. AB - The present article considers the influence of heterogeneity in a mobile analyte or in an immobilized ligand population on the surface binding kinetics and equilibrium isotherms. We describe strategies for solving the inverse problem of calculating two-dimensional distributions of rate and affinity constants from experimental data on surface binding kinetics, such as obtained from optical biosensors. Although the characterization of a heterogeneous population of analytes binding to uniform surface sites may be possible under suitable experimental conditions, computational difficulties currently limit this approach. In contrast, the case of uniform analytes binding to heterogeneous populations of surface sites is computationally feasible, and can be combined with Tikhonov-Phillips and maximum entropy regularization techniques that provide the simplest distribution that is consistent with the data. The properties of this ligand distribution analysis are explored with several experimental and simulated data sets. The resulting two-dimensional rate and affinity constant distributions can describe well experimental kinetic traces measured with optical biosensors. The use of kinetic surface binding data can give significantly higher resolution than affinity distributions from the binding isotherms alone. The shape and the level of detail of the calculated distributions depend on the experimental conditions, such as contact times and the concentration range of the analyte. Despite the flexibility introduced by considering surface site distributions, the impostor application of this model to surface binding data from transport limited binding processes or from analyte distributions can be identified by large residuals, if a sufficient range of analyte concentrations and contact times are used. The distribution analysis can provide a rational interpretation of complex experimental surface binding kinetics, and provides an analytical tool for probing the homogeneity of the populations of immobilized protein. PMID- 12770911 TI - Intrasequence GFP in class I MHC molecules, a rigid probe for fluorescence anisotropy measurements of the membrane environment. AB - Fluorescence anisotropy measurements can elucidate the microenvironment of a membrane protein in terms of its rotational diffusion, interactions, and proximity to other proteins. However, use of this approach requires a fluorescent probe that is rigidly attached to the protein of interest. Here we describe the use of one such probe, a green fluorescent protein (GFP) expressed and rigidly held within the amino acid sequence of a major histocompatibility complex (MHC) class I molecule, H2L(d). We contrast the anisotropy of this GFP-tagged MHC molecule, H2L(d)GFPout, with that of an H2L(d) that was GFP-tagged at its C terminus, H2L(d)GFPin. Both molecules fold properly, reach the cell surface, and are recognized by specific antibodies and T-cell receptors. We found that polarized fluorescence images of H2L(d)GFPout in plasma membrane blebs show intensity variations that depend on the relative orientation of the polarizers and the membrane normal, thus demonstrating that the GFP is oriented with respect to the membrane. These variations were not seen for H2L(d)GFPin. Before transport to the membrane surface, MHC class I associates with the transporter associated with antigen processing complex in the endoplasmic reticulum. The intensity dependent steady-state anisotropy in the ER of H2L(d)GFPout was consistent with FRET homotransfer, which indicates that a significant fraction of these molecules were clustered. After MCMV-peptide loading, which supplies antigenic peptide to the MHC class I releasing it from the antigen processing complex, the anisotropy of H2L(d)GFPout was independent of intensity, suggesting that the MHC proteins were no longer clustered. These results demonstrate the feasibility and usefulness of a GFP moiety rigidly attached to the protein of interest as a probe for molecular motion and proximity in cell membranes. PMID- 12770912 TI - A model for shear stress sensing and transmission in vascular endothelial cells. AB - Arterial endothelial cell (EC) responsiveness to flow is essential for normal vascular function and plays a role in the development of atherosclerosis. EC flow responses may involve sensing of the mechanical stimulus at the cell surface with subsequent transmission via cytoskeleton to intracellular transduction sites. We had previously modeled flow-induced deformation of EC-surface flow sensors represented as viscoelastic materials with standard linear solid behavior (Kelvin bodies). In the present article, we extend the analysis to arbitrary networks of viscoelastic structures connected in series and/or parallel. Application of the model to a system of two Kelvin bodies in parallel reveals that flow induces an instantaneous deformation followed by creeping to the asymptotic response. The force divides equally between the two bodies when they have identical viscoelastic properties. When one body is stiffer than the other, a larger fraction of the applied force is directed to the stiffer body. We have also probed the impact of steady and oscillatory flow on simple sensor-cytoskeleton nucleus networks. The results demonstrated that, consistent with the experimentally observed temporal chronology of EC flow responses, the flow sensor attains its peak deformation faster than intracellular structures and the nucleus deforms more rapidly than cytoskeletal elements. The results have also revealed that a 1-Hz oscillatory flow induces significantly smaller deformations than steady flow. These results may provide insight into the mechanisms behind the experimental observations that a number of EC responses induced by steady flow are not induced by oscillatory flow. PMID- 12770913 TI - Temporal variations in cell migration and traction during fibroblast-mediated gel compaction. AB - Current models used in our laboratory to assess the migration and traction of a population of cells within biopolymer gels are extended to investigate temporal changes in these parameters during compaction of mechanically constrained gels. The random cell migration coefficient, micro (t) is calculated using a windowing technique by regressing the mean-squared displacement of cells tracked at high magnification in three dimensions with a generalized least squares algorithm for a subset of experimental time intervals, and then shifting the window interval-by interval until all time points are analyzed. The cell traction parameter, tau(0)(t), is determined by optimizing the solution of our anisotropic biphasic theory to tissue equivalent compaction. The windowing technique captured simulated sinusoidal and step changes in cell migration superposed on a persistent random walk in simulated cell movement. The optimization software captured simulated time dependence of compaction on cell spreading. Employment of these techniques on experimental data using rat dermal fibroblasts (RDFs) and human foreskin fibroblasts (HFFs) demonstrated that these cells exhibit different migration-traction relationships. Rat dermal fibroblast migration was negatively correlated to traction, suggesting migration was not the driving force for compaction with these cells, whereas human foreskin fibroblast migration was positively correlated to traction. PMID- 12770914 TI - Structural-functional relationships of the dynein, spokes, and central-pair projections predicted from an analysis of the forces acting within a flagellum. AB - In the axoneme of eukaryotic flagella the dynein motor proteins form crossbridges between the outer doublet microtubules. These motor proteins generate force that accumulates as linear tension, or compression, on the doublets. When tension or compression is present on a curved microtubule, a force per unit length develops in the plane of bending and is transverse to the long axis of the microtubule. This transverse force (t-force) is evaluated here using available experimental evidence from sea urchin sperm and bull sperm. At or near the switch point for beat reversal, the t-force is in the range of 0.25-1.0 nN/ micro m, with 0.5 nN/ micro m the most likely value. This is the case in both beating and arrested bull sperm and in beating sea urchin sperm. The total force that can be generated (or resisted) by all the dyneins on one micron of outer doublet is also approximately 0.5 nN. The equivalence of the maximum dynein force/ micro m and t-force/ micro m at the switch point may have important consequences. Firstly, the t-force acting on the doublets near the switch point of the flagellar beat is sufficiently strong that it could terminate the action of the dyneins directly by strongly favoring the detached state and precipitating a cascade of detachment from the adjacent doublet. Secondly, after dynein release occurs, the radial spokes and central-pair apparatus are the structures that must carry the t-force. The spokes attached to the central-pair projections will bear most of the load. The central pair projections are well-positioned for this role, and they are suitably configured to regulate the amount of axoneme distortion that occurs during switching. However, to fulfill this role without preventing flagellar bend formation, moveable attachments that behave like processive motor proteins must mediate the attachment between the spoke heads and the central-pair structure. PMID- 12770915 TI - Unexpected mobility variation among individual secretory vesicles produces an apparent refractory neuropeptide pool. AB - Most stored neuropeptide cannot be released from nerve terminals suggesting the existence of a refractory pool of dense core vesicles (DCVs). Past fluorescence photobleaching recovery, single particle tracking and release experiments suggested that the refractory neuropeptide pool corresponds to a distinct immobile fraction of cytoplasmic DCVs. However, tracking of hundreds of individual green fluorescent protein-labeled neuropeptidergic vesicles by wide field or evanescent-wave microscopy shows that a separate immobile fraction is not evident. Instead, the DCV diffusion coefficient (D) distribution is unusually broad and asymmetric. Furthermore, the distribution shifts with a release facilitator. This unexpected variation, which could reflect heterogeneity among vesicles or in their medium, is shown to generate the appearance of a regulated refractory neuropeptide pool. PMID- 12770916 TI - Conformational dynamics and temperature dependence of photoinduced electron transfer within self-assembled coproporphyrin:cytochrome c complexes. AB - The focus of the present study is to better understand the complex factors influencing intermolecular electron transfer (ET) in biological molecules using a model system involving free-base coproporphyrin (COP) complexed with horse heart cytochrome c (Cc). Coproporphyrin exhibits bathochromic shifts in both the Soret and visible absorption bands in the presence of Cc and an absorption difference titration reveals a 1:1 complex with an association constant of 2.63 +/- 0.05 x 10(5) M(-1). At 20 degrees C, analysis of time-resolved fluorescence data reveals two lifetime components consisting of a discrete lifetime at 15.0 ns (free COP) and a Gaussian distribution of lifetimes centered at 2.8 ns (representing (1)COP -> Cc ET). Temperature-dependent, time-resolved fluorescence data demonstrate a shift in singlet lifetime as well as changes in the distribution width (associated with the complex). By fitting these data to semiclassical Marcus theory, the reorganizational energy (lambda) of the singlet state electron transfer was calculated to be 0.89 eV, consistent with values for other porphyrin/Cc intermolecular ET reactions. Using nanosecond transient absorption spectroscopy the temperature dependences of the forward and thermal back ET originating from triplet state were examined ((3)COP --> Cc ET). Fits of the temperature dependence of the rate constants to semiclassical Marcus theory gave lambda of 0.39 eV and 0.11 eV for the forward and back triplet ET, respectively (k(f) = (7.6 +/- 0.3) x 10(6) s(-1), k(b) = (2.4 +/- 0.3) x 10(5) s(-1)). The differing values of lambda for the forward and back triplet ET demonstrate that these ET reactions do not occur within a static complex. Comparing these results with previous studies of the uroporphyrin:Cc and tetrakis (4 carboxyphenyl)porphyrin:Cc complexes suggests that side-chain flexibility gives rise to the conformational distributions in the (1)COP --> Cc ET whereas differences in overall porphyrin charge regulates gating of the back ET reaction (reduced Cc --> COP(+)). PMID- 12770918 TI - Na/K pump current and [Na](i) in rabbit ventricular myocytes: local [Na](i) depletion and Na buffering. AB - Na/K pump current (I(pump)) and intracellular Na concentration ([Na](i)) were measured simultaneously in voltage-clamped rabbit ventricular myocytes, under conditions where [Na](i) is controlled mainly by membrane transport. Upon abrupt pump reactivation (after 10-12 min blockade), I(pump) decays in two phases. Initially, I(pump) declines with little [Na](i) change, whereas the second phase is accompanied by [Na](i) decline. Initial I(pump) sag was still present at external [K] = 15 mM, but prevented by [Na](i) approximately 100 mM. Initial I(pump) sag might be explained by subsarcolemmal [Na](i) ([Na](SL)) depletion produced by rapid Na extrusion and I(pump). Brief episodes of pump blockade allowed [Na](SL) repletion, since peak postblockade I(pump) exceeded I(pump) at the end of previous activation (without appreciably altered global [Na](i)). The apparent K(m) for [Na](i) was higher for continuous I(pump) activation than peak I(pump) (14.1 +/- 0.2 vs. 11.2 +/- 0.2 mM), whereas that based on d[Na](i)/dt matched peak I(pump) (11.6 +/- 0.3 mM). [Na](SL) depletion (vs. [Na](i)) could be as high as 3 mM for [Na](i) approximately 18-20 mM. A simple diffusion model indicates that such [Na](SL) depletion requires a Na diffusion coefficient 10(3)- to 10(4)-fold below that expected in bulk cytoplasm (although this could be subsarcolemmal only). I(pump) integrals and [Na](i) decline were used to estimate intracellular Na buffering, which is slight (1.39 +/- 0.09). PMID- 12770919 TI - Tonic and phasic receptor neurons in the vertebrate olfactory epithelium. AB - Olfactory receptor neurons (ORNs) respond to odorants with characteristic patterns of action potentials that are relevant for odor coding. Prolonged odorant exposures revealed three populations of dissociated toad ORNs, which were mimicked by depolarizing currents: tonic (TN, displaying sustained firing, 49% of 102 cells), phasic (PN, exhibiting brief action potential trains, 36%) and intermediate neurons (IN, generating trains longer than PN, 15%). We studied the biophysical properties underlying the differences between TNs and PNs, the most extreme cases among ORNs. TNs and PNs possessed similar membrane capacitances (approximately 4 pF), but they differed in resting potential (-82 versus -64 mV), input resistance (4.2 versus 2.9 G(Omega)) and unspecific current, I(u) (TNs: 0 < I(u) 1 pA/pF). Firing behavior did not correlate with differences in voltage-gated conductances. We developed a mathematical model that accurately simulates tonic and phasic patterns. Whole cell recordings from rat ORNs in fragments (approximately 4 mm(2)) of olfactory epithelium showed that such a tissue normally contains tonic and phasic receptor neurons, suggesting that this feature is common across a wide range of vertebrates. Our findings show that the individual passive electrical properties can govern the firing patterns of ORNs. PMID- 12770917 TI - Activation of phospholipase C increases intramembrane electric fields in N1E-115 neuroblastoma cells. AB - We imaged the intramembrane potential (a combination of transmembrane, surface, and dipole potential) on N1E-115 neuroblastoma cells with a voltage-sensitive dye. After activation of the B(2) bradykinin receptor, the electric field sensed by the dye increased by an amount equivalent to a depolarization of 83 mV. The increase in intramembrane potential was blocked by the phospholipase C (PLC) inhibitors U-73122 and neomycin, and was invariably accompanied by a transient rise of [Ca(2+)](i). A depolarized inner surface potential, as the membrane loses negative charges via phosphatidylinositol 4,5-bisphosphate (PIP(2)) hydrolysis, and an increase in the dipole potential, as PIP(2) is hydrolyzed to 1,2 diacylglycerol (DAG), can each account for a small portion of the change in intramembrane potential. The primary contribution to the measured change in intramembrane potential may arise from an increased dipole potential, as DAG molecules are generated from hydrolysis of other phospholipids. We found bradykinin produced an inhibition of a M-type voltage-dependent K(+) current (I(K(M))). This inhibition was also blocked by the PLC inhibitors and had similar kinetics as the bradykinin-induced modulation of intramembrane potential. Our results suggest that the change in the local intramembrane potential induced by bradykinin may play a role in mediating the I(K(M)) inhibition. PMID- 12770920 TI - Hydrodynamic recruitment of rolling leukocytes in vitro. PMID- 12770922 TI - Colitis: it is not just for the colon anymore. PMID- 12770923 TI - Do fluorescent drugs show you more than you wanted to know? PMID- 12770925 TI - The neuromedin B receptor antagonist, BIM-23127, is a potent antagonist at human and rat urotensin-II receptors. AB - The functional activity of the peptidic neuromedin B receptor antagonist BIM 23127 was investigated at recombinant and native urotensin-II receptors (UT receptors). Human urotensin-II (hU-II) promoted intracellular calcium mobilization in HEK293 cells expressing the human UT (hUT) or rat UT (rUT) receptors with pEC(50) values of 9.80+/-0.34 (n=6) and 9.06+/-0.32 (n=4), respectively. While BIM-23127 alone had no effect on calcium responses in either cell line, it was a potent and competitive antagonist at both hUT (pA(2)=7.54+/ 0.14; n=3) and rUT (pA(2)=7.70+/-0.05; n=3) receptors. Furthermore, BIM-23127 reversed hU-II-induced contractile tone in the rat-isolated aorta with a pIC(50) of 6.66+/-0.04 (n=4). In conclusion, BIM- 23127 is the first hUT receptor antagonist identified to date and should not be considered as a selective neuromedin B receptor antagonist. PMID- 12770924 TI - Brain renin-angiotensin system dysfunction in hypertension: recent advances and perspectives. AB - This review focuses on the dysfunction of the intrinsic brain renin-angiotensin system (RAS) in the pathogenesis of hypertension. Hyperactivity of the brain RAS plays a critical role in mediating hypertension in both humans and animal models of hypertension, including the spontaneously hypertensive rat (SHR). The specific mechanisms by which increased brain RAS activity results in hypertension are not well understood but include increases in sympathetic vasomotor tone and impaired arterial baroreflex function. We discuss the contribution of endogenous angiotensin (Ang) II actions on presympathetic vasomotor rostral ventrolateral medulla neurons to enhance sympathetic activity and maintain hypertension. In addition, we discuss Ang II-induced attenuation of afferent baroreceptor feedback within the nucleus tractus solitarius and its relevance to the development of hypertension. We also outline the cellular and molecular mechanisms of Ang II signal transduction that may be critical for the initiation and establishment of hypertension. In particular, we present evidence for a phosphoinositide-3-kinase dependent signaling pathway that appears to contribute to hypertension in the SHR, possibly via augmented Ang II-induced increases in neuronal firing rate and enhanced transcriptional noradrenaline neuromodulation. Finally, we outline future directions in utilizing our understanding of the brain RAS dysfunction in hypertension for the development of improved therapeutic intervention in hypertension. PMID- 12770926 TI - Curcumin, the major component of food flavour turmeric, reduces mucosal injury in trinitrobenzene sulphonic acid-induced colitis. AB - 1 Inflammmatory bowel disease (IBD) is characterized by oxidative and nitrosative stress, leucocyte infiltration and upregulation of proinflammatory cytokines. In this study, we have investigated the protective effects of curcumin, an anti inflammatory and antioxidant food derivative, on 2,4,6- trinitrobenzene sulphonic acid-induced colitis in mice, a model for IBD. 2 Intestinal lesions (judged by macroscopic and histological score) were associated with neutrophil infiltration (measured as increase in myeloperoxidase activity in the mucosa), increased serine protease activity (may be involved in the degradation of colonic tissue) and high levels of malondialdehyde (an indicator of lipid peroxidation). 3 Dose response studies revealed that pretreatment of mice with curcumin (50 mg kg(-1) daily i.g. for 10 days) significantly ameliorated the appearance of diarrhoea and the disruption of colonic architecture. Higher doses (100 and 300 mg kg(-1)) had comparable effects. 4 In curcumin-pretreated mice, there was a significant reduction in the degree of both neutrophil infiltration (measured as decrease in myeloperoxidase activity) and lipid peroxidation (measured as decrease in malondialdehyde activity) in the inflamed colon as well as decreased serine protease activity. 5 Curcumin also reduced the levels of nitric oxide (NO) and O(2)(-) associated with the favourable expression of Th1 and Th2 cytokines and inducible NO synthase. Consistent with these observations, nuclear factor-kappaB activation in colonic mucosa was suppressed in the curcumin-treated mice. 6 These findings suggest that curcumin or diferuloylmethane, a major component of the food flavour turmeric, exerts beneficial effects in experimental colitis and may, therefore, be useful in the treatment of IBD. PMID- 12770927 TI - Thymidine phosphorylase induces angiogenesis in vivo and in vitro: an evaluation of possible mechanisms. AB - 1 Thymidine phosphorylase (TP) is elevated in the plasma of cancer patients, and has been implicated in pathophysiological angiogenesis. However, the downstream signals underlying this implication remain obscure. The purpose of the present study was to examine the effects of TP on the neovascularisation response in vitro and in vivo. 2 Both TP and its catalytic product, 2-deoxy-D-ribose-1 phosphate, and downstream 2-deoxy-D-ribose (2-DDR) promoted endothelial tubulogenesis in vitro, and the regeneration of a wounded monolayer of endothelial cells without exerting any mitogenic effect. In vivo, both TP and 2 DDR promoted the development of functional vasculature into an avascular sponge. A TP inhibitor, 6-amino-5-chlorouracil, was able to partially reverse the effects of TP, but had no effect on the 2-DDR-induced angiogenesis. 3 Enhanced monolayer regeneration was observed with TP-cDNA-transfected bladder carcinoma cells. The transfection of TP-cDNA, however, did not confer any proliferative advantage. The regeneration of TP overexpressing cells was associated with a time-dependent expression of the enzyme haeme-oxygenase (HO-1). 4 The present study demonstrates that both TP and its ribose-sugar metabolites induce angiogenesis by mediating a cohesive interplay between carcinoma and endothelial cells. The induction of HO-1 in TP-transfected cells suggests that it could be a possible downstream signal for the angiogenic effects of TP. Furthermore, reducing sugars have been shown to induce oxidative stress, and ribose could be a possible cause for the upregulation of HO-1, which has been implicated in the release of angiogenic factors. Therefore, we postulate that 2-DDR could be mediating the angiogenic effects of TP possibly through an oxidative stress mechanism and additionally getting integrated in the endothelial metabolic machinery. PMID- 12770928 TI - Pharmacology and direct visualisation of BODIPY-TMR-CGP: a long-acting fluorescent beta2-adrenoceptor agonist. AB - 1 Fluorescence techniques offer a way to circumvent several problems associated with many radioligand binding and functional assays and the need for large numbers of cells. Fluorescent ligands also offer the advantage of allowing real time direct visualisation of ligand - receptors interactions. A fluorescent analogue of CGP 12177 (BODIPY-TMR-CGP) has thus been evaluated as a beta(2) adrenoceptor ligand in CHO-K1 cells expressing the human beta(2)-adrenoceptor. 2 Studies of (3)H-cAMP accumulation showed that BODIPY-TMR-CGP stimulated an increase in cAMP accumulation and cyclic AMP response element (CRE)-mediated gene transcription with an EC(50) of 21-28 nM. Both of these responses were antagonised by the selective beta(2)-adrenoceptor antagonist ICI 118551. 3 Binding studies with (3)H-CGP 12177, and functional studies of CRE-regulated gene transcription showed that the BODIPY-TMR-CGP interaction with the human beta(2) adrenoceptor is of very long duration. 4 Visualisation of the binding of BODIPY TMR-CGP to single living mammalian cells was clearly demonstrated by confocal microscopy and showed that this ligand was able to selectively label cell surface beta(2)-adrenoceptors in living CHO-K1 cells transfected with the human beta(2) adrenoceptor with an apparent K(D) of 27 nM. Studies with cells expressing a beta(2)-adrenoceptor-green fluorescent protein (GFP) fusion protein provided further strong evidence that BODIPY-TMR-CGP was binding to the beta(2) adrenoceptor. 5 BODIPY-TMR-CGP is therefore a long-acting fluorescent beta(2) adrenoceptor agonist that can be used to label beta(2)-adrenoceptors in the plasma membrane of living cells. PMID- 12770929 TI - 5-HT decreases contractile and electrical activities in lymphatic vessels of the guinea-pig mesentery: role of 5-HT 7-receptors. AB - 1 Constriction measurements and intracellular microelectrode recordings were performed in vitro on lymphatic vessels isolated from the guinea-pig mesentery to investigate whether 5-hydroxytryptamine (5-HT) affected lymphatic pumping and smooth muscle membrane potential. 2 5-HT decreased in a concentration-dependent manner the frequency of constrictions induced by intraluminal vessel perfusion. In nonperfused vessels, 5-HT hyperpolarized the lymphatic smooth muscle membrane potential and decreased the frequency and amplitude of spontaneous transient depolarizations (STDs). 3 The actions of 5-HT were significantly reversed by the 5-HT(7) receptor antagonist (2R)-1-[(3-hydroxyphenyl)sulfonyl]-2-[2-(4-methyl-1 piperidinyl)ethyl]pyrrolidine (SB269970, 0.5 micro M) and by the 5-HT(1/2/5/7) receptor antagonists methysergide (0.5 micro M), and were mimicked by the 5 HT(1/7)-receptor agonist, 5-CT. 4 The 5-HT(4)-receptor antagonists 1-methyl-1H indole-3-carboxylic acid [1-2-[(methyl sulfonyl) amino] ethyl-4-piperidinyl] methyl ester (GR113808, 1 micro M) and (1-piperidinyl) ethyl 1H-indole 3 carboxylate (SB203186, 1 micro M) did not significantly affect the 5-HT-induced responses. The 5-HT(4)-receptor agonist 1-(4-amino-5-chloro-2-methoxy-phenyl)-3 [1-(2-methylsulfonylamino) ethyl-4-piperidinyl]-1-propanone hydrochloride (RS67506) decreased the constriction frequency, albeit only at 50 micro M and without affecting the smooth muscle membrane potential. 5 Responses to 5-HT were attenuated by the nitric oxide synthase inhibitor N(G)-nitro L-arginine (100 micro M), whereas indomethacin (10 micro M) and tetrodotoxin (1 micro M) were without effects. 6 5-HT-induced responses were inhibited by the ATP-sensitive K(+) channel blocker, glibenclamide (10 micro M) and the cAMP-dependent protein kinase inhibitor N-[2-(p-bromociannamylamino)-ethyl]-5-isoquinolinesulfonamide dichloride (H89, 10 micro M) blocked the hyperpolarization. 7 These results suggest that 5-HT modulates the rate of lymphatic vessel pumping by eliciting K(ATP) channel-mediated smooth muscle hyperpolarization and decrease in STD activity, which appear to be mediated by activation of 5-HT(7) receptors coupled to cAMP production. PMID- 12770930 TI - Dualistic actions of cromakalim and new potent 2H-1,4-benzoxazine derivatives on the native skeletal muscle K ATP channel. AB - 1 New 2H-1,4-benzoxazine derivatives were synthesized and tested for their agonist properties on the ATP-sensitive K(+) channels (K(ATP)) of native rat skeletal muscle fibres by using the patch-clamp technique. The novel modifications involved the introduction at position 2 of the benzoxazine ring of alkyl substituents such as methyl (-CH(3)), ethyl (-C(2)H(5)) or propyl ( C(3)H(7)) groups, while maintaining pharmacophore groups critical for conferring agonist properties. 2 The effects of these molecules were compared with those of cromakalim in the presence or absence of internal ATP (10(-4) M). In the presence of internal ATP, all the compounds increased the macropatch K(ATP) currents. The order of potency of the molecules as agonists was -C(3)H(7) (DE(50)=1.63 x 10(-8) M) >-C(2)H(5) (DE(50)=1.11 x 10(-7) M)>-CH(3) (DE(50)=2.81 x 10(-7) M)>cromak slim (DE(50)= 1.42 x 10(-5) M). Bell-shaped dose-response curves were observed for these compounds and cromakalim indicating a downturn in response when a certain dose was exceeded. 3 In contrast, in the absence of internal ATP, all molecules including cromakalim inhibited the K(ATP) currents. The order of increasing potency as antagonists was cromakalim (IC(50)=1.15 x 10(-8) M)> or = CH(3) (IC(50)=2.6 x 10(-8) M)>-C(2)H(5) (IC(50)=4.4 x 10(-8) M)>-C(3)H(7) (IC(50)=1.68 x 10(-7) M) derivatives. 4 These results suggest that the newly synthesized molecules and cromakalim act on muscle K(ATP) channel by binding on two receptor sites that have opposite actions. Alternatively, a more simple explanation is to consider the existence of a single site for potassium channel openers regulated by ATP which favours the transduction of the channel opening. The alkyl chains at position 2 of the 2H-1,4-benzoxazine nucleus is pivotal in determining the potency of benzoxazine derivatives as agonists or antagonists. PMID- 12770931 TI - Low endotoxemia prevents the reduction of gastric blood flow induced by NSAIDs: role of nitric oxide. AB - 1 The role of nitric oxide (NO) in the effects of low endotoxemia on gastric damage and blood flow has been evaluated in indomethacin-treated rats. 2 Pretreatment (-1 h) with endotoxin (40 micro g kg(-1)) reduced gastric damage induced by indomethacin (20 mg kg(-1)) in conscious rats. 3 Endotoxin prevented the reduction in gastric blood flow (laser Doppler flowmetry) induced by indomethacin in pentobarbital-anaesthetised rats. 4 Pretreatment with an NO synthase (NOS) inhibitor (L-NAME, 1 mg kg(-1)) reversed the protective effect of endotoxin on gastric blood perfusion. 5 Endotoxin did not modify the expression of mRNA for endothelial NOS or inducible NOS in the gastric corpus when evaluated 1 h postinjection. However, a 3.8-fold increase in inducible NOS mRNA and a 61% reduction in endothelial NOS mRNA were observed in the gastric corpus 4 h after endotoxin administration. 6 Evaluation of both total and Ca(2+)-dependent NOS activity by analysing the rate of conversion of L-arginine to L-citrulline in gastric corpus homogenates showed no differences between animals treated with endotoxin and those treated with saline 1 or 4 h beforehand. Ca(2+)-independent NOS activity was almost non-apparent in control as well as in endotoxin-treated rats at all the time points analysed. 7 Low endotoxemia preserves blood perfusion and protects the gastric mucosa against the deleterious effects of indomethacin through the endogenous NO release. NO synthesis in response to endotoxin does not involve the inducible NOS, but probably depends on the post translational/biochemical regulation in vivo of a Ca(2+)-dependent NOS, most probably endothelial NOS. PMID- 12770932 TI - Effect of a kinin B2 receptor antagonist on LPS- and cytokine-induced neutrophil migration in rats. AB - 1 This study examines the involvement of kinins in neutrophil migration into rat subcutaneous air pouches triggered by lipopolysaccharide (LPS), as well as the putative roles played by kinin B(1) and B(2) receptors, tumour necrosis factor alpha (TNF-alpha), interleukin-1 beta (IL-1beta) and selectins in this response. 2 LPS (5 ng to 10 micro g cavity(-1)) injected into the 6-day-old pouch induced a dose- and time-dependent neutrophil migration which peaked between 4 and 6 h, and was maximal following the dose of 100 ng cavity(-1) (saline: 0.46+/-0.1; LPS: 43+/-3.70 x 10(6) cells cavity(-1) at 6 h). 3 Bradykinin (BK) (600 nmol) injected into the pouch of saline-treated rats induced only modest neutrophil migration (0.73+/-0.16 x 10(6) cells cavity(-1)). A more robust response to BK (3.2+/-0.25 x 10(6) cells cavity(-1)) was seen in animals pretreated with captopril, but this was still smaller than the responses to IL-1beta or TNF-alpha (15 pmol: 23+/-2.2 x 10(6) and 75 pmol: 29.5+/-2 x 10(6) cells cavity(-1), respectively). Nevertheless, the B(1) agonist des-Arg(9)-BK (600 nmol) failed to induce neutrophil migration. 4 HOE-140 (1 and 2 mg kg(-1)), a B(2) receptor antagonist, reduced LPS-induced neutrophil migration. HOE-140 also reduced the neutrophil migration induced by BK, but had no effect on the migration promoted by IL-1beta or TNF-alpha. des-Arg(9)-[Leu(8)]-BK, B(1) receptor antagonist was ineffective in changing neutrophil migration caused by any of these stimuli. 5 Neutrophil migration induced by LPS or BK was reduced by interleukin-1 receptor antagonist (IL-1ra) (1 mg kg(-1)), sheep anti-rat TNF serum (anti-TNF serum) (0.3 ml cavity( 1)), and the nonspecific selectin inhibitor fucoidin (10 mg kg(-1)). 6 TNF-alpha levels in the pouch fluid were increased by LPS or BK injection, peaking at 0.5-1 h and gradually declining thereafter up to 6 h. IL-1beta levels increased steadily throughout the 6 h period. HOE-140 markedly inhibited the rise in IL 1beta and TNF-alpha levels in pouch fluid triggered by both stimuli. 7 These results indicate that BK participates importantly in selectin-dependent neutrophil migration into the air pouch triggered by LPS in the rat, by stimulating B(2) receptors coupled to synthesis/release of TNF-alpha and IL 1beta. PMID- 12770933 TI - P2Y receptor regulation of cultured rat cerebral cortical cells: calcium responses and mRNA expression in neurons and glia. AB - 1 We have investigated increases in cytosolic Ca(2+) in response to nucleotides in mixed rat cerebrocortical cultures (neurons and glia in similar numbers) and in essentially neuron-free glial cultures. 2 In both cultures, the agonist response profile was 2-methylthioADP(2MeSADP)>2 methylthioATP(2MeSATP)>ADP>ATP>adenosine 5'-O-(3-thiotriphosphate), consistent with a P2Y(1) receptor. The maximal responses to 2MeSADP, 2MeSATP and ADP were identical, but that to ATP was higher. 3 Suramin, pyridoxal-phosphate-6-azophenyl 2',4'-disulphonic acid, reactive blue 2 (RB2), and adenosine biphosphate (A3P5P) were antagonists with apparent pA(2) values of 5.5 for suramin, 6.4 for RB2, and 4.7 for A3P5P. 4 Single cell imaging divided the cells from the mixed neuronal glial cultures into two populations: responsive (neurons) and unresponsive (glial cells) to high [K(+)]. The response of cells to nucleotides was almost exclusively limited to those not responsive to high K(+). 5 In the presence of extracellular Mn(2+), the response of the mixed cultures to 30 mM K(+) and 20 micro M Bay K 8644 was attenuated. However, when 2MeSADP was added there was no reduction in response in cultures previously loaded with Mn(2+). This further indicated that the 2MeSADP response was not in the neurons. 6 Reverse transcriptase-polymerase chain reaction studies detected transcripts for P2Y(1), P2Y(4) and P2Y(6) in RNA preparations from embryonic rat cortex, and from both mixed and glial cultures. P2Y(2) transcripts were not detected in the embryonic cortex. 7 Based on this and previous work, it is proposed that the principal P2Y influences in the brain are on cytosolic Ca(2+) in glial cells and presynaptic sites on neurons. PMID- 12770934 TI - Signaling mechanisms of enhanced neutrophil phagocytosis and chemotaxis by the polysaccharide purified from Ganoderma lucidum. AB - 1 The polysaccharide from Ganoderma lucidum (PS-G) has been reported to enhance immune responses and to elicit antitumor effects. In our previous study, we found that PS-G efficiently inhibited spontaneously and Fas-enhanced neutrophil apoptosis when cultured in vitro. Since phagocytosis and chemotaxis play essential roles in host defense mediated by neutrophils, it is of great interest to know the effect of PS-G on these two cell functions, and the molecular events leading to these actions. 2 Using latex beads and heat-inactive Escherichia coli serving as particles for neutrophil engulfment, we found that PS-G is able to enhance phagocytic activity of human primary neutrophils and neutrophilic phenotype cells differentiated from all trans retinoic acid-treated HL-60 cells. 3 Chemotactic assay using Boyden chamber also revealed the ability of PS-G to increase neutrophil migration. 4 Exposure of neutrophils to PS-G time dependently caused increases in protein kinase C (PKC), p38 mitogen-activated protein kinase (MAPK), Hck, and Lyn activities. 5 Results with specific kinase inhibitors indicate that phagocytic action of PS-G was reduced by the presence of wortmannin (Phosphatidylinositol 3-kinase, PI3K inhibitor), pyrazolpyrimidine 2 (Src-family tyrosine kinase inhibitor), Ro318220 (PKC inhibitor), and SB203580 (p38 MAPK inhibitor), but not by PD98059 (mitogen-activated protein/ERK kinase inhibitor). Moreover, chemotactic action of PS-G requires the activities of PI3K, p38 MAPK, Src tyrosine kinases and PKC. 6 All these results demonstrate the abilities of PS G to enhance neutrophil function in phagocytosis and chemotaxis, and further provide evidence to strengthen the beneficial remedy of G. lucidum in human to enhance defense system. PMID- 12770935 TI - Mechanism of kinin release during experimental acute pancreatitis in rats: evidence for pro- as well as anti-inflammatory roles of oedema formation. AB - 1 Kinin B(2) receptor antagonists or tissue kallikrein (t-KK) inhibitors prevent oedema formation and associated sequelae in caerulein-induced pancreatitis in the rat. We have now further investigated the mechanism of kinin generation in the pancreas. 2 Kinins were elevated in the pancreatic tissue already before oedema formation became manifest. Peak values (421+/-59 pmol g(-1) dry wt) were reached at 45 min and remained elevated for at least 2 h; a second increase was observed at 24 h. Pretreatment with the B(2) receptor antagonist icatibant abolished kinin formation, while post-treatment was ineffective. 3 Total kininogen levels were very low in the pancreas of controls, but increased 75-fold during acute pancreatitis. This increase was absent in rats that were pretreated with icatibant. 4 During pancreatitis, t-KK-like and plasma kallikrein (p-KK)-like activity in the pancreas, as well as trypsinogen activation peptide (TAP) increased significantly. Icatibant pretreatment further augmented t-KK about 100 fold, while p-KK was significantly attenuated; TAP levels remained unaffected. 5 Endogenous protease inhibitors (alpha(1)-antitrypsin, alpha(2)-macroglobulin) were low in normal tissues, but increased 45- and four-fold, respectively, during pancreatitis. This increase was abolished when oedema formation was prevented by icatibant. 6 In summary, oedema formation is initiated by t-KK; the ensuing plasma protein extravasation supplies further kininogen and active p-KK to the tissue. Concomitantly, endogenous protease inhibitors in the oedema fluid inhibit up to 99% of active t-KK. Our data thus suggest a complex interaction between kinin action and kinin generation involving positive and negative feedback actions of the inflammatory oedema. PMID- 12770936 TI - Altered prejunctional modulation of intestinal cholinergic and noradrenergic pathways by alpha2-adrenoceptors in the presence of experimental colitis. AB - 1 This study investigates the influence of intestinal inflammation on: (1) the control of intestinal neurotransmission and motility by prejunctional alpha(2) adrenoceptors and (2) the expression of intestinal alpha(2)-adrenoceptors. Experimental colitis was induced by intrarectal administration of 2,4 dinitrobenzenesulphonic acid (DNBS) to rats. 2 UK-14,304 inhibited atropine sensitive electrically evoked contractions of ileal and colonic longitudinal muscle preparations. UK-14,304 acted with similar potency, but higher efficacy, on tissues from DNBS-treated animals; its effects were antagonized with greater potency by phentolamine than rauwolscine. 3 Electrically induced [(3)H]noradrenaline release from ileal preparations was reduced in the presence of colitis. Tritium outflow was decreased by UK-14,304 and stimulated by rauwolscine or phentolamine: these effects were enhanced in preparations from animals with colitis. 4 Reverse transcription-polymerase chain reaction and Western blot assay demonstrated the protein expression of alpha(2A)-adrenoceptors in mucosal and muscular tissues isolated from ileum and colon. The induction of colitis increased alpha(2A)-adrenoceptor expression in both ileal and colonic muscular layers, without concomitant changes in mucosal tissues. 5 Induction of colitis reduced gastrointestinal propulsion of a charcoal suspension in vivo. In this setting, the gastrointestinal transit was inhibited by intraperitoneal (i.p.) UK-14,304 and stimulated by i.p. rauwolscine. After pretreatment with guanethidine, the stimulant action of rauwolscine no longer occurred, and UK 14,304 exerted a more prominent inhibitory effect that was antagonized by rauwolscine. 6 The present results indicate that, in the presence of intestinal inflammation, prejunctional alpha(2)-adrenoceptors contribute to an enhanced inhibitory control of cholinergic and noradrenergic transmission both at inflamed and noninflamed distant sites. Evidence was obtained that such modulatory actions depend on an increased expression of alpha(2A)-adrenoceptors within the enteric nervous system. PMID- 12770937 TI - Mechanisms involved in the stimulation of prostacyclin synthesis by human lymphocytes in human umbilical vein endothelial cells. AB - 1 Endothelial cells play an important role in the modulation of vascular tone because of their ability to produce vasoactive substances such as prostacyclin (PGI(2)). Cell-cell contact between human umbilical vein endothelial cells (HUVEC) and peripheral blood lymphocytes has been shown to stimulate endothelial PGI(2) synthesis by increasing free arachidonic acid availability through endothelial cytosolic phospholipase A2 (cPLA(2)) activation. In this study, we sought to determine whether phospholipase C (PLC) and D (PLD) activation also contributes, besides cPLA(2), to the lymphocyte-induced PGI(2) synthesis in HUVEC, and to delineate further the potential mechanisms of cPLA(2) activation triggered by the interaction of HUVEC with lymphocytes. 2 Pretreatment of endothelial cells with the PI-PLC inhibitor U-73122 before the coincubation with lymphocytes markedly inhibited the PGI(2) output whereas the diacylglycerol (DAG) lipase inhibitor RHC 80267 and ethanol had no effect. These results suggest that PLC may be involved through inositol trisphosphate generation and calcium mobilization, and that neither DAG nor phosphatidic acid (PtdOH) was used as sources of arachidonic acid. 3 The stimulated PGI(2) synthesis was protein kinase C (PKC)-independent but strongly inhibited by the mitogen-activated protein kinase kinase (MEK) inhibitors PD98059 and U-0126 and by the Src kinase inhibitor PP1. 4 Immunoblot experiments showed an increased phosphorylation of the extracellular signal-regulated kinases 1/2 (ERK1/2) upon lymphocyte addition till 4 h coincubation. Phosphorylation was markedly inhibited by U-0126 and PP1 addition. 5 Collectively, these results suggest that the signaling cascade triggered by lymphocytes in endothelial cells involves an Src kinase/ERK1/2 pathway leading to endothelial cPLA(2) activation. PMID- 12770939 TI - Receptor signaling mechanisms underlying muscarinic agonist-evoked contraction in guinea-pig ileal longitudinal smooth muscle. AB - 1 In guinea-pig ileal longitudinal muscle, muscarinic partial agonists, 4-(N-[3 chlorophenyl]-carbomoyloxy)-2-butynyl-trimethylammonium (McN-A343) and pilocarpine, each produced parallel increases in tension and cytosolic Ca(2+) concentration ([Ca(2+)]c) with a higher EC(50) than that of the full agonist carbachol. The maximum response of [Ca(2+)]c or tension was not much different among the three agonists. The Ca(2+) channel blocker nicardipine markedly inhibited the effects of all three agonists 2 The contractile response to any agonist was antagonized in a competitive manner by M(2) receptor selective antagonists (N,N'-bis[6-[[(2-methoyphenyl)methyl]amino]hexyl]-1,8-octanediamine tetrahydrochloride and 11-[[2-[(diethlamino)methyl]-1-piperidinyl]acetyl]-5,11 dihydro-6H-pyrido[2,3-b][1,4] benzodiazepine-6-one), and the apparent order of M(2) antagonist sensitivity was McN-A343>pilocarpine>carbachol. M(3) receptor selective antagonists, 1,1-dimethyl-4-diphenylacetoxypiperidinium iodide and darifenacin, both severely depressed the maximum response for McN-A343, while darifenacin had a similar action in the case of pilocarpine. Both M(3) antagonists behaved in a competitive manner in the case of the carbachol response. 3 McN-A343 failed to release Ca(2+) from the intracellular stores, and the Ca(2+)-releasing action of pilocarpine was very weak compared with that of carbachol. All three agonists were capable of increasing Ca(2+) sensitivity of the contractile proteins. 4 McN-A343 rarely produced membrane depolarization, but always accelerated electrical spike discharge. Pilocarpine effect was more often accompanied by membrane depolarization, as was usually seen using carbachol. 5 The results suggest that muscarinic agonist-evoked contractions result primarily from the integration of Ca(2+) entry associated with the increased spike discharge and myofilaments Ca(2+) sensitization, and that Ca(2+) store release may contribute to the contraction indirectly via potentiation of the electrical membrane responses. They may also support the idea that an interaction of M(2) and M(3) receptors plays a crucial role in mediating the contraction response. PMID- 12770938 TI - Angiopoietin-1 inhibits endothelial permeability, neutrophil adherence and IL-8 production. AB - 1 Angiopoietin-1 (Ang1) is an angiogenic growth factor that binds to the Tie2 receptor on vascular endothelium, promoting blood vessel maturation and integrity. In the present study, we have investigated whether Ang1 also possesses anti-inflammatory properties by determining its effects on endothelial barrier function, neutrophil (PMN) adherence to endothelial cells (EC) and production of the PMN chemotactic factor interleukin-8 (IL-8). 2 Pretreatment of endothelial monolayers with Ang1 attenuated the permeability increase induced by thrombin in both lung microvascular cells and a human endothelial cell line. Similarly, Ang1 prevented the permeability-inducing effects of platelet-activating factor, bradykinin and histamine. 3 Pretreatment of EC with Ang1 also reduced the adherence of PMN to EC stimulated by thrombin. In contrast to its ability to counteract the increase in monolayer permeability brought about by various inflammatory agents, Ang1 did not affect the ability of histamine, PAF, or tumor necrosis factor-alpha to stimulate PMN adherence to EC. 4 In addition to its ability to inhibit PMN adherence, Ang1 diminished IL-8 production from EC challenged with thrombin in a concentration-dependent manner. 5 When EC were preincubated with the specific Rho kinase (ROCK) inhibitor Y-27632, we observed a reduction in PMN adherence in response to thrombin, as well as a decrease in thrombin-stimulated IL-8 production. Coincubation of monolayers with Y-27632 and Ang1 did not further attenuate the above-mentioned responses. However, Ang-1 failed to inhibit the activation of RhoA in response to thrombin, suggesting that inhibition of EC adhesiveness for PMN and IL-8 production by Ang1 does not result from reduced ROCK activation. 6 We conclude that Ang1 can counteract several aspects of the inflammatory response, including endothelial permeability, PMN adherence to EC as well as inhibition of IL-8 production by EC. PMID- 12770940 TI - Loperamide mobilizes intracellular Ca2+ stores in insulin-secreting HIT-T15 cells. AB - 1 We have investigated the effects of loperamide on intracellular Ca(2+) stores and membrane K(+) channels in insulin-secreting hamster insulinoma (HIT-T15) cells. 2 In cell-attached patch-clamp mode, loperamide (3-250 micro M) activated large single-channel currents. The loperamide-activated currents were tentatively identified as Ca(2+)-activated K(+) channel (K(Ca)) currents based on their single-channel conductance (145 pS), apparent reversal potential, and insensitivity to tolbutamide. Smaller single-channel currents with a conductance (32 pS) indicative of adenosine triphosphate-sensitive K(+) channels (K(ATP) channels) were also recorded, but were insensitive to loperamide. 3 Surprisingly, the loperamide-activated currents persisted in the absence of extracellular Ca(2+). Yet under these conditions, we still measured loperamide-induced Ca(2+) increases. These effects are dose dependent. Loperamide had no effects in the inside-out patch configuration, suggesting that loperamide does not directly activate the channels with large conductance, but does so secondarily to release of Ca(2+) from intracellular stores. 4 Carbachol (100 micro M), an agonist of muscarinic receptors, which mediates IP(3)-dependent intracellular Ca(2+) release, enhanced the effects of loperamide on K(Ca) channels. 5 Both the putative K(Ca) currents and Ca(2+) signals induced by loperamide (with '0' [Ca(2+)](o)) were abolished when the intracellular Ca(2+) stores had been emptied by pretreating the cells with either carbachol or thapsigargin, an endoplasmic reticulum Ca(2+)-ATPase inhibitor that blocks reuptake of calcium. 6 These data indicate that loperamide in insulin-secreting beta-cells evokes intracellular Ca(2+) release from IP(3)-gated stores and activates membrane currents that appear to be carried by K(Ca), rather than K(ATP) channels. PMID- 12770941 TI - Cellular actions of opioids on periaqueductal grey neurons from C57B16/J mice and mutant mice lacking MOR-1. AB - 1 Patch clamp recordings were made from periaqueductal grey (PAG) neurons in vitro to investigate the cellular actions of opioids in wild-type C57B16/J mice and mutant mice lacking the first exon of the micro -opioid (MOP) receptor. 2 In wild-type mice, the kappa-(KOP) agonist U-69593 (300 nM) and the mixed micro /delta-opioid agonist met-enkephalin (10 micro M), but not the delta-(DOP) agonist deltorphin (300 nM), reduced the amplitude of evoked GABA(A)-mediated inhibitory postsynaptic currents (IPSCs). Met-enkephalin and U-69593 also reduced the rate of spontaneous miniature IPSCs, but had no effect on their amplitude and kinetics. In micro -receptor-deleted mice, only U-69593 (300 nM) reduced the amplitude of evoked IPSCs. 3 In wild-type mice, the MOP agonist DAMGO (3 micro M) produced an outward current in 76% of the neurons. Deltorphin and U-69593 produced outward currents in 24 and 32% of the neurons, respectively. In micro receptor-deleted mice, deltorphin and U-69593 produced similar outward currents in 32 and 27% of the neurons, respectively, while DAMGO was without effect. All neurons in both the wild-type and micro -receptor-deleted mice responded with similar outward currents to either the GABA(B) receptor agonist baclofen (10 micro M), or the opioid-like receptor ORL1 (NOP) agonist nociceptin (300 nM). 4 The DAMGO-, deltorphin-, U-69593-, baclofen- and nociceptin-induced currents displayed inward rectification and reversed polarity at -109 to -116 mV. 5 These findings indicate that micro -, delta- and kappa-opioid receptor activation has complex pre- and postsynaptic actions within the mouse PAG. This differs to the rat PAG where only micro -opioid receptor actions have been observed. PMID- 12770943 TI - Effects of methyl p-hydroxybenzoate (methyl paraben) on Ca2+ concentration and histamine release in rat peritoneal mast cells. AB - 1 Mechanisms of methyl p-hydroxybenzoate (methyl paraben) action in allergic reactions were investigated by measuring the intracellular Ca(2+) concentration ([Ca(2+)](i)) and histamine release in rat peritoneal mast cells (RPMCs). 2 In the presence or absence of extracellular Ca(2+), methyl paraben (0.1-10 mM) increased [Ca(2+)](i), in a concentration-dependent manner. Under both the conditions, methyl paraben alone did not evoke histamine release. 3 In RPMCs pretreated with a protein kinase C (PKC) activator (phorbol 12-myristate 13 acetate (PMA) 3 and 10 nM), methyl paraben (0.3-3 mM) induced histamine release. However, a high concentration (10 mM) of the agent did not increase the histamine release. 4 U73122 (0.1 and 0.5 micro M), an inhibitor of phospholipase C (PLC), significantly inhibited the methyl paraben-induced histamine release in PMA pretreated RPMCs. U73343 (0.5 micro M), an inactive analogue of U73122, did not inhibit the histamine release caused by methyl paraben. 5 In Ca(2+)-free solution, PLC inhibitors (U73122 0.1 and 0.5 micro M, D609 1-10 micro M) inhibited the methyl paraben-induced increase in [Ca(2+)](i), whereas U73343 (0.5 micro M) did not. 6 Xestospongin C (2-20 micro M) and 2 aminoethoxydiphenyl borate (30 and 100 micro M), blockers of the inositol 1,4,5-trisphosphate (IP(3)) receptor, inhibited the methyl paraben-induced increase in [Ca(2+)](i) in Ca(2+) free solution. 7 In conclusion, methyl paraben causes an increase in [Ca(2+)](i), which may be due to release of Ca(2+) from storage sites by IP(3) via activation of PLC in RPMCs. In addition, methyl paraben possibly has some inhibitory effects on histamine release via unknown mechanisms. PMID- 12770942 TI - Binding and effect of K ATP channel openers in the absence of Mg2+. AB - 1 Openers of ATP-sensitive K(+) channels (K(ATP) channels) are thought to act by enhancing the ATPase activity of sulphonylurea receptors (SURs), the regulatory channel subunits. At higher concentrations, some openers activate K(ATP) channels also in the absence of MgATP. Here, we describe binding and effect of structurally diverse openers in the absence of Mg(2+) and presence of EDTA. 2 Binding of openers to SUR2B was measured using a mutant with high affinity for [(3)H]glibenclamide ([(3)H]GBC). In the absence of Mg(2+), 'typical' openers (benzopyrans, cyanoguanidines and aprikalim) inhibited [(3)H]GBC binding with K(i) values approximately 200 x higher than in the presence of MgATP. Minoxidil sulphate and nicorandil were inactive, whereas binding of diazoxide was unaffected by MgATP. 3 In the absence/presence of MgATP, N-cyano-N'-(1,1 dimethylpropyl)-N"-3-pyridylguanidine (P1075) activated the Kir6.2/SUR2B channel in inside-out patches with EC(50)=2000/67nM and E(max)=32/134%. In the absence of Mg(2+), responses were variable with only a small part of the variability being explained by a decrease in channel responsiveness with time after patch excision and to differences in the ATP sensitivity between patches. 4 The rank order of efficacy of the openers was P1075>rilmakalim approximately nicorandil>diazoxide>minoxidil sulphate. 5 The data show that structurally diverse openers are able to bind to, and to activate the Kir6.2/SUR2B channel by a pathway independent of ATP hydrolysis. These effects are observed at concentrations used to define the biochemical mechanism of the openers in the presence of MgATP and allow the openers to be classified into 'typical' and 'atypical' KCOs with diazoxide standing apart. PMID- 12770944 TI - Role of leukotrienes in the regulation of human granulocyte behaviour: dissociation between agonist-induced activation and retardation of apoptosis. AB - 1 Since most inflammatory mediators that stimulate granulocyte responsiveness also delay apoptosis, it is often assumed that activation and longevity are causally related. Using isolated human peripheral blood neutrophils and eosinophils, we examined this association by exploiting the proinflammatory lipid mediators, the leukotrienes (LTs), and investigated granulocyte function and apoptosis. 2 LTB(4) induced elevation of intracellular free Ca(2+) concentration ([Ca(2+)](i)), cell polarisation and retardation of neutrophil apoptosis, although the antiapoptotic effect occurred only at concentrations > or =300 nM. LTB(4)-induced activation was attenuated by CP-105,696, a BLT1-specific antagonist suggesting classical LTB(4) receptor BLT1 involvement. 3 Despite demonstrating the presence of the neutrophil intracellular LTB(4) receptor peroxisome-proliferator activator receptor-alpha (PPARalpha) in neutrophils, the selective PPARalpha agonist WY-14,643 did not mimic LTB(4)-induced prosurvival effects. 4 LTB(4)-induced survival, however, also appeared to be mediated by BLT1 since CP-105,696 inhibited the LTB(4)-mediated antiapoptotic effect. Furthermore, based on studies with CP-105,696 and 5-lipoxygenase inhibitors, lipopolysaccharide (LPS)-, granulocyte-macrophage colony-stimulating factor (GM CSF)-, dexamethasone- and dibutyryl-cAMP (db-cAMP)-induced delay of neutrophil apoptosis did not involve autocrine production of LTB(4). 5 Although LTB(4) and LTD(4) induced human eosinophil [Ca(2+)](i) elevation and polarization, these LTs did not influence eosinophil apoptosis. Furthermore, LTB(4)- and LTD(4)-induced eosinophil activation was attenuated by CP-105,696 and the Cys-LT(1) receptor antagonist montelukast, respectively, highlighting specific receptor dependency. 6 Thus, mediator-triggered granulocyte activation and antiapoptotic pathways are distinct events that can be differentially regulated. PMID- 12770945 TI - Castration decreases amylase release associated with muscarinic acetylcholine receptor downregulation in rat parotid gland. AB - 1 The mechanism and receptor subtypes involved in carbachol-stimulated amylase release and its changes after castration were studied in parotid slices from male rats. 2 Carbachol induced both amylase release and inositol phosphate (IP) accumulation in parotid slices from control and castrated rats, but castration induced a decrease of carbachol maximal effect. The effect of castration was reverted by testosterone replacement. 3 The selective M(1) and M(3) muscarinic receptor antagonists, pirenzepine and 4-diphenylacetoxy-N-methylpiperidine methiodide, respectively, inhibited carbachol-stimulated amylase release and IP accumulation in a dose-dependent manner in parotid slices from control and castrated rats. 4 A diminution of binding sites of muscarinic receptor in parotid membrane from castrated rats was observed. Competition binding assays showed that both, M(1) and M(3) muscarinic receptor subtypes are expressed in membranes of parotid glands from control and castrated rats, M(3) being the greater population. 5 These results suggest that amylase release induced by carbachol in parotid slices is mediated by phosphoinositide accumulation. This mechanism appears to be triggered by the activation of M(1) and M(3) muscarinic receptor subtypes. Castration induced a decrease of the maximal effect of carbachol evoked amylase release and IP accumulation followed by a diminution in the number of parotid gland muscarinic acetylcholine receptors. PMID- 12770946 TI - Conventional-type protein kinase C contributes to phorbol ester-induced inhibition of rat myometrial tension. AB - 1 Phorbol ester decreases muscle tension in the rat myometrium, and the effect is more potent in late-pregnant myometrium than in nonpregnant myometrium. In the present study, we have examined the contribution of protein kinase C (PKC) isoforms to the phorbol ester-induced inhibition of tension in rat uterine smooth muscle. 2 Thymeleatoxin (THX), a selective activator of conventional-type PKC (cPKC), and 12-deoxyphorbol 13-isobutyrate (DPB), an activator of pan PKC, inhibited the tension induced by high K(+), and inhibitions were significantly increased in pregnant myometrium compared to nonpregnant myometrium. The inhibition by DPB and THX of high K(+)-induced tension was significantly attenuated when PKC was downregulated by long-term pretreatment with THX and inhibited by Go6976, a cPKC inhibitor. 3 Of the cPKCs, PKC alpha is predominantly expressed in the rat myometrium, as detected by Western blot analysis. The expression of PKC alpha gradually increases from the beginning of gestation, reaching a maximum at day 21 of pregnancy. Treatment with DPB induced PKC alpha to translocate from the cytosol to the membrane in the pregnant myometrium. PKC epsilon and PKC zeta, other dominant PKC isoforms in the rat myometrium, decrease during gestation, reaching a minimum in late pregnancy. 4 These results suggest that cPKC may be at least partly involved in the PKC-mediated inhibition of muscle tension in the rat myometrium. PMID- 12770948 TI - Dihydroergotamine and its metabolite, 8'-hydroxy-dihydroergotamine, as 5-HT1A receptor agonists in the rat brain. AB - 1 In addition to stopping migraine attacks, dihydroergotamine (DHE) is an efficient drug for migraine prophylaxis. Whether 5-HT(1A) receptors could contribute to the latter action was assessed by investigating the effects of DHE and its metabolite, 8'-OH-DHE, on these receptors in the rat brain. 2 Membrane binding assays with [(3)H]8-OH-DPAT and [(3)H]WAY 100635 as radioligands showed that both DHE (IC(50)=28-30 nM) and 8'-OH-DHE (IC(50)=8-11 nM) are high-affinity 5-HT(1A) receptor ligands. 3 Both DHE and 8'-OH-DHE enhanced the specific binding of [(35)S]GTP-gamma-S to the dorsal raphe nucleus and the hippocampus in brain sections, but to a lower extent than 5-carboxamido-tryptamine (5-CT) in the latter area. 4 Both DHE (EC(50)=10.9+/-0.3 nM) and 8'-OH-DHE (EC(50)=30.4+/-0.8 nM) inhibited the firing of serotoninergic neurons in the dorsal raphe nucleus within brain stem slices. 5 Intracellular recording showed that 8'-OH-DHE was more potent than DHE to hyperpolarize CA1 pyramidal cells in rat hippocampal slices. 6 Both the stimulatory effects of DHE and 8'-OH-DHE on [(35)S]GTP-gamma-S binding and their electrophysiological effects were completely prevented by the selective 5-HT(1A) receptor antagonist WAY 100635. 7 As expected of 5-HT(1A) receptor partial agonists, DHE and 8'-OH-DHE prevented any subsequent hyperpolarization of CA1 pyramidal cells by 5-HT or 5-CT. 8 Through their actions at 5-HT(1A) auto- (in the dorsal raphe nucleus) and hetero-(notably in the hippocampus) receptors, DHE, and even more its metabolite 8'-OH-DHE, can exert both an inhibitory influence on neuronal excitability and anxiolytic effects which might contribute to their antimigraine prophylactic efficiency. PMID- 12770947 TI - Roles for CCK1 and 5-HT3 receptors in the effects of CCK on presympathetic vasomotor neuronal discharge in the rat. AB - 1 The role of peripheral 5-hydroxytryptamine (5-HT(3)) receptors and cholecystokinin type 1 (CCK(1)) receptors in the inhibitory effects of phenylbiguanide (PBG) and CCK on arterial blood pressure, heart rate and the discharge of presympathetic vasomotor neurones of the rostral ventrolateral medulla (RVLM) was studied in alpha-chloralose-anaesthetized rats. 2 CCK (1 and 4 micro g kg(-1), i.v.) and PBG (2 and 10 micro g kg(-1), i.v.) reduced arterial blood pressure and heart rate, and inhibited the discharge of single RVLM presympathetic vasomotor neurones in a dose-related manner. 3 Devazepide (0.5 mg kg(-1), i.v.), a selective CCK(1) receptor antagonist, blocked the effects of CCK on arterial blood pressure, heart rate and neuronal discharge but did not significantly alter these responses to PBG. MDL72222 (0.1 mg kg(-1), i.v.), a selective 5-HT(3) receptor antagonist, blocked the effects of PBG on arterial blood pressure, heart rate and presympathetic neuronal discharge. MDL72222 attenuated the effects of CCK on arterial blood pressure, heart rate and RVLM presympathetic neuronal discharge. Vehicle did not significantly alter any of the responses to CCK or PBG. 4 These experiments suggest that systemically administered CCK acts directly through CCK(1) receptors to modulate sympathetic vasomotor function. In addition, the actions of CCK also are partly dependent on activation of 5-HT(3) receptors. CCK may release 5-HT which then acts at 5-HT(3) receptors to produce sympathetic vasomotor inhibition. In contrast, the actions of PBG are entirely dependent on 5-HT(3) receptors and are independent of any actions at the CCK(1) receptor. PMID- 12770949 TI - A comparison of Ca2+ channel blocking mode between gabapentin and verapamil: implication for protection against hypoxic injury in rat cerebrocortical slices. AB - 1 The mode of Ca(2+) channel blocking by gabapentin [1-(aminomethyl)cyclohexane acetic acid] was compared to those of other Ca(2+) channel blockers, and the potential role of Ca(2+) channel antagonists in providing protection against hypoxic injury was subsequently investigated in rat cerebrocortical slices. 2 mRNA for the alpha(2)delta subunits of Ca(2+) channels was found in rat cerebral cortex. 3 Nitric oxide (NO) synthesis estimated from cGMP formation was enhanced by KCl stimulation, which was mediated primarily by the activation of N- and P/Q type Ca(2+) channels. Gabapentin blocked both types of Ca(2+) channels, and preferentially reversed the response to 30 mM K(+) stimulation compared with 50 mM K(+) stimulation. In contrast, verapamil preferentially inhibited the response to depolarization by the higher concentration (50 mM) of K(+). 4 Gabapentin inhibited KCl-induced elevation of intracellular Ca(2+) in primary neuronal culture. 5 Hypoxic injury was induced in cerebrocortical slices by oxygen deprivation in the absence (severe injury) or presence of 3 mM glucose (mild injury). Gabapentin preferentially inhibited mild injury, while verapamil suppressed only severe injury. omega-Conotoxin GVIA (omega-CTX) and omega agatoxin IVA (omega-Aga) were effective in both models. 6 NO synthesis was enhanced in a manner dependent on the severity of hypoxic insults. Gabapentin reversed the NO synthesis induced by mild insults, while verapamil inhibited that elicited by severe insults. omega-CTX and omega-Aga were effective in both the cases. 7 Therefore, the data suggest that gabapentin and verapamil cause activity dependent Ca(2+) channel blocking by different mechanisms, which are associated with their cerebroprotective actions and are dependent on the severity of hypoxic insults. PMID- 12770950 TI - Involvement of H2O2 in superoxide-dismutase-induced enhancement of endothelium dependent relaxation in rabbit mesenteric resistance artery. AB - 1 The mechanism underlying the enhancement by superoxide dismutase (SOD) of endothelium-dependent relaxation was investigated in rabbit mesenteric resistance arteries. 2 SOD (200 U ml(-1)) increased the production of H(2)O(2) in smooth muscle cells (as indicated by the use of an H(2)O(2)-sensitive fluorescent dye). 3 Neither SOD nor catalase (400 U ml(-1)) modified either the resting membrane potential or the hyperpolarization induced by acetylcholine (ACh, 1 micro M) in smooth muscle cells. 4 In arteries constricted with noradrenaline, the endothelium-dependent relaxation induced by ACh (0.01-1 micro M) was enhanced by SOD (200 U ml(-1)) (P<0.01). This action of SOD was inhibited by L-N(G) nitroarginine (nitric oxide (NO)-synthase inhibitor) but not by either charybdotoxin+apamin (Ca(2+)-activated-K(+)-channel blockers) or diclofenac (cyclooxygenase inhibitor). 5 Neither ascorbate (50 micro M) nor tiron (0.3 mM), superoxide scavengers, had any effect on the ACh-induced relaxation, but each attenuated the enhancing effect of SOD on the ACh-induced relaxation. Similarly, catalase (400 U ml(-1)) inhibited the effect of SOD without changing the ACh induced relaxation. 6 In endothelium-denuded strips constricted with noradrenaline, SOD enhanced the relaxation induced by the NO donor 1-hydroxy-2 oxo-3-(N-methyl-3-aminopropyl)-3-methyl-1-triazene (NOC-7) (P<0.05). Ascorbate and catalase each attenuated this effect of SOD. 7 H(2)O(2) (1 micro M) enhanced the relaxation on the noradrenaline contraction induced by NOC-7 and that induced by 8-bromo-cGMP, a membrane-permeable analogue of guanosine 3',5' cyclic monophosphate (cGMP). 8 SOD had no effect on cGMP production, whether measured in endothelium-intact strips following an application of ACh (0.1 micro M) or in endothelium-denuded strips following an application of NOC-7 (0.1 micro M). 9 It is suggested that in rabbit mesenteric resistance arteries, SOD increases the ACh induced, endothelium-dependent relaxation by enhancing the action of NO in the smooth muscle via its H(2)O(2)-producing action (rather than via a superoxide scavenging action). PMID- 12770951 TI - Serotonin upregulates the activity of phagocytosis through 5-HT1A receptors. AB - 1 In this study, we investigated whether serotonin could regulate the in vitro activity of phagocytosis through 5-hydroxytryptamine or serotonin (5-HT(1A)) receptors. 2 Mouse peritoneal macrophages were cultured with serotonin and the activity of phagocytosis was assessed by the uptake of zymosan and latex particles added to the culture media. Specific binding of [(3)H]8-OH-DPAT and immunohistochemistry using an affinity-purified anti-5-HT(1A)-receptor antibody were assayed in the macrophages. In addition, we took advantage of the availability of pharmacological inhibitors of nuclear factor-kappaB (NF-kappaB) to explore its role in the regulation of the 5-HT(1A) receptor. 3 Serotonin increased the in vitro activity of phagocytosis in a dose-dependent manner. The 5 HT(1A) receptor agonist (+/-)-8-hydroxy-2-(di-n-propyl-amino)-tetralin (R(+)-8-OH DPAT) reproduced these effects. Serotonin- or R(+)-8-OH-DPAT-induced increases in phagocytosis were blocked by the 5-HT(1A) receptor antagonist WAY100635 and the NF-kappaB inhibitor pyrrolidinedithiocarbamate. Moreover, mouse peritoneal macrophages expressed specific binding sites for [(3)H]8-OH-DPAT when cultivated in the presence of zymosan or latex beads. Immunohistochemistry confirmed the expression of the 5-HT(1A) receptor protein in the macrophages. 4 These results show that serotonin can upregulate the activity of peritoneal macrophages through 5-HT(1A) receptors. PMID- 12770952 TI - Deletion of the UT receptor gene results in the selective loss of urotensin-II contractile activity in aortae isolated from UT receptor knockout mice. AB - 1 Urotensin-II (U-II) is among the most potent mammalian vasoconstrictors identified and may play a role in the aetiology of essential hypertension. Currently, only one mouse U-II receptor (UT) gene has been cloned. It is postulated that this protein is solely responsible for mediating U-II-induced vasoconstriction. 2 This hypothesis has been investigated in the present study, which assessed basal haemodynamics and vascular reactivity to hU-II in wild-type (UT((+/+))) and UT receptor knockout (UT((-/-))) mice. 3 Basal left ventricular end-diastolic and end-systolic volumes/pressures, stroke volumes, mean arterial blood pressures, heart rates, cardiac outputs and ejection fractions in UT((+/+)) mice and in UT((-/-)) mice were similar. 4 Relative to UT((+/+)) mouse isolated thoracic aorta, where hU-II was a potent spasmogen (pEC(50)=8.26+/-0.08) that evoked relatively little vasoconstriction (17+/-2% 60 mM KCl), vessels isolated from UT((-/-)) mice did not respond to hU-II. However, in contrast, the superior mesenteric artery isolated from both the genotypes did not contract in the presence of hU-II. Reactivity to unrelated vasoconstrictors (phenylephrine, endothelin-1, KCl) and endothelium-dependent/independent vasodilator agents (carbachol, sodium nitroprusside) was similar in the aorta and superior mesenteric arteries isolated from both the genotypes. 5 The present study is the first to directly link hU-II-induced vasoconstriction with the UT receptor. Deletion of the UT receptor gene results in loss of hU-II contractile action with no 'nonspecific' alterations in vascular reactivity. However, as might be predicted based on the limited contractile efficacy recorded in vitro, the contribution that hU-II and its receptor make to basal systemic haemodynamics appears to be negligible in this species. PMID- 12770953 TI - Heavenly rings. PMID- 12770954 TI - Central serous chorioretinopathy. PMID- 12770955 TI - Second generation vital stains in retinal surgery. PMID- 12770956 TI - Too dry or not too dry. PMID- 12770957 TI - Extracapsular cataract surgery compared with manual small incision cataract surgery in community eye care setting in western India: a randomised controlled trial. AB - AIM: To study "manual small incision cataract surgery (MSICS)" for the rehabilitation of cataract visually impaired and blind patients in community based, high volume, eye hospital setting; to compare the safety and effectiveness of MSICS with conventional extracapsular cataract surgery (ECCE). METHODS: In a single masked randomised controlled clinical trial, 741 patients, aged 40-90 years, with operable cataract were randomly assigned to receive either MSICS or ECCE and operated upon by one of eight participating surgeons. Intraoperative and postoperative complications were graded and scored according to the Oxford Cataract Treatment and Evaluation Team recommendations. The patients were followed up at 1 week, 6 weeks, and 1 year after surgery and their visual acuity recorded. RESULTS: This paper reports outcomes at 1 and 6 weeks. 706 of the 741(95.3%) patients completed the 6 week follow up. 135 of 362 (37.3%) of ECCE group and 165 of 344 (47.9%) of MSICS group had uncorrected visual acuity of 6/18 or better after 6 weeks of follow up. 314 of 362 (86.7%) of ECCE group and 309 of 344 (89.8%) of MSICS group had corrected postoperative vision of 6/18 or better. Four of 362 (1.1%) of ECCE group and six of 344 (1.7%) of MSICS group had corrected postoperative visual acuity less than 6/60. There were no significant differences between the two groups for intraoperative and severe postoperative complications. CONCLUSION: MSICS and ECCE are both safe and effective techniques for treatment of cataract patients in community eye care settings. MSICS needs similar equipment to ECCE, but gives better uncorrected vision. PMID- 12770958 TI - The Armenian Eye Care Project: surgical outcomes of complicated paediatric glaucoma. AB - AIM: To review surgical results of the Armenian Eye Care Project's (AECP) efforts in paediatric glaucoma and to identify additional morbidity associated with its initial itinerant nature. METHODS: A non-randomised, retrospective historical cohort study of complicated paediatric glaucoma surgeries was performed by the AECP at the Republican Eye Hospital. 38 eyes of 34 paediatric patients underwent mitomycin C trabeculectomies (MMCT) (n = 20) or Ahmed glaucoma valve (AGV) placements (n = 18) for complicated paediatric glaucomas. The median age was 12.5 years with a range of 6-17 years. Three clinical groups were represented: congenital glaucoma, Frank Kamenetsky syndrome (a glaucoma associated with uveitis), and Rieger's syndrome. RESULTS: Preoperative and postoperative intraocular pressures were similar for the two groups (MMCT v AGV = 30 v 32 mm Hg and 13.5 v 14.8 mm Hg, respectively; p>0.05, t test). The AGV group was twice as likely to use medications postoperatively (44% v 23%; p<0.05, t test). The MMCT group was more likely to have a three line drop in visual acuity than the AGV group (28% v 12%; p<0.05, t test). The decreased visual acuity found postoperatively was most commonly secondary to cataract formation. CONCLUSION: Total success rates were similar in the two surgical treatment groups. These compared favourably with success rates reported for non-itinerant surgery. Postoperative complications worsened by the itinerant nature of the project were few and represented 5% of the total surgical interventions. Ultimately, the installation of fellowship trained specialists is the most effective approach to minimising this problem and is recommended by the AECP as a strategy for developing and recovering countries. PMID- 12770959 TI - Low vision and blindness in adults in Gurage Zone, central Ethiopia. AB - AIM: To determine the magnitude and causes of low vision and blindness in the Gurage zone, central Ethiopia. METHODS: A cross sectional study using a multistage cluster sampling technique was used to identify the study subjects. Visual acuity was recorded for all adults 40 years and older. Subjects who had a visual acuity of <6/18 were examined by an ophthalmologist to determine the cause of low vision or blindness. RESULTS: From the enumerated population, 2693 (90.8%) were examined. The prevalence of blindness (<3/60 better eye presenting vision) was 7.9% (95% CI 6.9 to 8.9) and of low vision (6/24-3/60 better eye presenting vision) was 12.1% (95% CI 10.9 to 13.3). Monocular blindness was recorded in 16.3% of the population. Blindness and low vision increased with age. The odds of low vision and blindness in women were 1.8 times that of the men. The leading causes of blindness were cataract (46.1%), trachoma (22.9%), and glaucoma (7.6%). While the prevalence of vision reducing cataract increased with age, the prevalence of trachoma related vision loss did not increase with age, suggesting that trichiasis related vision loss in this population might not be cumulative. CONCLUSION: The magnitude of low vision and blindness is high in this zone and requires urgent intervention, particularly for women. Further investigation of the pattern of vision loss, particularly as a result of trachomatous trichiasis, is warranted. PMID- 12770960 TI - A clinical follow up of PRK and LASIK in eyes with preoperative abnormal corneal topographies. AB - AIM: To assess the safety and predictability of photorefractive keratotomy (PRK) and laser in situ keratomileusis (LASIK) based on preoperative corneal topography. METHODS: A non-randomised comparative study was carried out on 84 eyes that presented with topographic abnormalities before undergoing PRK (n = 44) or LASIK (n = 40) procedures. 84 spherical equivalent paired normal eyes served as the control group. Either PRK or LASIK procedures were performed on 168 eyes using the Summit apex plus excimer laser. Topographic abnormalities, including apex displacement (AD), increased asphericity (AS), meridional irregularity (MI), increased inferior-superior asymmetry (IS), increased curvature (CU), and combined features (CO), were assessed preoperatively using the EyeSys analysis system. Safety and predictability of the two procedures were defined as a postoperative visual acuity of 20/40 or better and the loss of one or more lines of spectacle corrected visual acuity (SCVA). RESULTS: All patients were followed for 6 months. There was a significant loss of best corrected visual acuity in the PRK-AD (p<0.001), PRK-CO (p<0.05), and LASIK-AS (p<0.001) patients. The number of eyes within plus or minus 1.0D of the surgical plan postoperatively was similar in all groups. CONCLUSION: These data suggest that although predictability was similar, PRK and LASIK performed in corneas with topographic abnormalities might cause loss of vision. PMID- 12770961 TI - H626R and R124C mutations of the TGFBI (BIGH3) gene caused lattice corneal dystrophy in Vietnamese people. AB - BACKGROUND/AIMS: Mutations of the human transforming growth factor beta induced gene (TGFBI) were reported to cause lattice corneal dystrophy (LCD) in various nationalities. This study analysed the TGFBI gene in Vietnamese people with LCD. METHODS: 13 unrelated families, including 34 patients and 21 unaffected members were examined. 50 normal Vietnamese people served as controls. Blood samples were collected. Genomic DNA was extracted from leucocytes. Analysis of TGFBI gene was performed using the polymerase chain reaction and direct sequencing. Corneal buttons were studied histopathologically. RESULTS: Two clinically distinguishable forms of LCD were revealed: one was typical of LCDI; the other was characterised by the late onset, thick lattice lines, and asymmetry between two eyes. Sequencing of the TGFBI gene revealed R124C mutation in three families and H626R mutation in 10 families. Congo red staining of the H626R-LCD cornea showed amyloid deposits in the subepithelial and stromal layers. CONCLUSIONS: R124C and H626R mutations of TGFBI gene caused LCD in Vietnamese people. R124C, a common cause of LCDI in many nationalities, was relatively rare, whereas H626R reported in several white people but not yet in Asians was most common (>75%) in Vietnamese people. Since the phenotype caused by H626R represents a new variant intermediate between LCDI and LCDIIIA, we proposed to consider it as LCD type IIIB. PMID- 12770962 TI - A comparison of perimetric results with the Medmont and Humphrey perimeters. AB - BACKGROUND: The Humphrey field analyser (HFA), Humphrey-Zeiss frequency doubling perimeter, and the Medmont automated perimeter (MAP) are three commonly used automated perimeters with threshold achromatic methodologies. Visual field loss may be detected earlier with strategies that target cell lines with reduced redundancy or which suffer selective damage. METHOD: To compare these three perimeters, 63 subjects who were glaucoma suspects, ocular hypertensives, glaucoma patients, or normal controls were recruited selectively. All subjects underwent testing using MAP central threshold, MAP flicker perimetry, HFA full threshold, HFA SITA perimetry, HFA short wavelength perimetry (SWAP), and frequency doubling perimetry (FDP). After visual field testing, equivalent tests were compared: MAP central threshold with HFA full threshold and HFA SITA perimetry; Medmont flicker perimetry with HFA SWAP and FDP. RESULTS: On analysis of the MAP central threshold a kappa statistic and an area under the receiver operator curve (AUC) of 0.90 and 0.94, respectively, were found compared with HFA full threshold strategies, and 0.87 and 0.92 respectively, compared with HFA SITA. For MAP flicker a kappa statistic and an AUC of 0.65 and 0.81, respectively, were found compared with HFA SWAP and 0.87 and 0.96, respectively, compared with FDP. A quadrant analysis and comparison of mean defect between tests was also highly significant. CONCLUSION: Medmont and Humphrey perimeters correlated well; both may be used for clinical and research purposes with similar confidence. PMID- 12770963 TI - Comparison of localised nerve fibre layer defects in normal tension glaucoma and primary open angle glaucoma. AB - AIM: To compare the pattern of localised nerve fibre layer (NFL) defects in normal tension glaucoma (NTG) and primary open angle glaucoma (POAG). METHODS: 50 NTG eyes and 36 POAG eyes, all with localised NFL defects, were enrolled. On retinal NFL photography, the proximity of the defect to the centre of the fovea (angle alpha) and the sum of the angular width of the defects (angle beta) were determined. Angle alpha was the angle made by a line from the centre of the fovea to the disc centre and a line from the disc centre to the disc margin, where the nearest border of the defect met. The patterns of localised NFL defects in NTG and POAG were compared with angles alpha and beta. Independent t test was used for statistical analysis. RESULTS: Angle alpha in NTG (35.1 (SD 20.0) degrees ) was significantly smaller than that of POAG (45.9 (21.9) degrees ) (p=0.02), while angle beta in NTG (49.0 (31.9) degrees ) was significantly larger than that of POAG (33.1 (23.9) degrees ) (p=0.01). CONCLUSIONS: The pattern of NFL defects in NTG was different from that in POAG. Localised NFL defects in NTG were closer to the fovea and wider in width than those in POAG. PMID- 12770964 TI - Transcaruncular approach for the management of frontoethmoid mucoceles. AB - AIMS: To present transcaruncular medial orbitotomy as the preferred approach to manage frontoethmoid mucoceles. METHODS: 11 patients with frontoethmoid mucoceles received transcaruncular orbitotomy from 2000 to 2002 at the National Taiwan University Hospital. The incision was made through the caruncle to explore the medial wall periosteum. Then the periosteum was opened and extended to provide adequate surgical field exposure. Frontoethmoid mucoceles could be viewed and removed directly. A transnasal drainage tube was inserted before closure of the caruncle wound. RESULTS: The mean follow up period was 12 (SD 9.1) months (range 1-26). Both functional recovery and cosmetic outcome were excellent. There was no recurrence of mucoceles. One patient complained of diplopia, which subsided after 2 months. CONCLUSION: Transcaruncular orbitotomy provides a wide exposure and a safe access to the medial orbital extraperiosteal space. Furthermore, there is less damage to skin and muscle layer and less manipulation of other ocular adnexal structures, such as medial canthal tendon and lacrimal sac. It can be an excellent approach for the management of frontoethmoid mucoceles. PMID- 12770965 TI - Psychosomatic aspects in patients with central serous chorioretinopathy. AB - AIMS: Patients with central serous chorioretinopathy were tested for psychosomatic symptoms and social support at the onset of their ailments; their personality profiles were also tested. METHODS: 24 consecutive outpatients at the department of ophthalmology, Freiburg University Hospital, Freiburg, Germany, presenting with central serous chorioretinopathy filled in the following standardised questionnaires: Symptom Checklist 90-R (SCL 90-R), Symptom List, Questionnaire on Social Support, and Sixteen Personality Factor Questionnaire. RESULTS: The sample (n=24) included 22 male patients with an average age of 44.1 years. Three quarters of the patients have children and live with their families; three quarters are white collar workers or self employed; one quarter are skilled blue collar workers. 12 patients were suffering from the ailment for the first time; 12 patients were undergoing a relapse. Mean visus of the affected eye was 0.8. The Symptom List yielded an inconspicuous total score for psychosomatic symptoms. In the SCL 90-R, nine out of 24 patients (37%) evinced elevated psychic stress. The total value of the questionnaire on social support showed good social support. In the Sixteen Personality Factor Questionnaire, the patients showed heightened emotional instability and insecurity as well as flexibility and spontaneity. CONCLUSION: The sociodemographic data confirm that central serous chorioretinopathy is predominantly an ailment of middle aged, socially well integrated men. This study suggests elevated psychic stress a few weeks after the onset of the ailment. The personality factors in the patients with central serous chorioretinopathy found in this study have to be verified with other populations. These results did not clearly show that psychosocial factors have a definite role in the aetiology of central serous chorioretinopathy. In order to make any clear assertions about this matter, further longitudinal studies on the progression of psychosocial parameters with larger patient samples would be indispensable. PMID- 12770966 TI - Features of abnormal choroidal circulation in central serous chorioretinopathy. AB - AIMS: To evaluate abnormalities in the choroidal circulation in cases of central serous chorioretinopathy (CSC). METHODS: A complete clinical ophthalmological examination was performed using simultaneous fluorescein and indocyanine green (ICG) angiography with a confocal scanning laser ophthalmoscopy and the digital images analysed in 36 consecutive patients with acute CSC. To quantify the choroidal circulation, the foveal choroidal blood flow was measured in 11 patients using laser Doppler flowmetry. RESULTS: Fluorescein angiography showed focal leakage from the retinal pigment epithelium in all patients. ICG angiography revealed delays in arterial filling in 27 eyes (75%), and fluorescein angiography showed small hypofluorescent points around the leakage in 27 eyes (75%). Abnormal choroidal hyperfluorescence was observed in 30 eyes (83%). The choroidal blood flow in eyes with CSC was 45% lower than in fellow eyes (p<0.01). CONCLUSION: Decreased choroidal blood flow in CSC was demonstrated for the first time. The decreased choroidal blood flow might be correlated with the small, localised hypofluorescent areas, which may indicate non-perfused areas of the choriocapillaris that are frequently seen during ICG angiography. PMID- 12770967 TI - Double vital staining using trypan blue and infracyanine green in macular pucker surgery. AB - AIMS: To study the clinical properties of double vital staining in premacular fibrosis, facilitating complete removal of all epiretinal tissue. METHODS: In a two step surgery, the epiretinal pucker was removed after staining with trypan blue, whereafter the inner limiting membrane was peeled after staining with infracyanine green. RESULTS: In all 30 patients, a separate epiretinal layer and inner limiting membrane were removed from the macular area. Pathological examination showed different histological properties of the removed layers. An increased visual acuity was measured in 26 patients, and a slightly decreased visual acuity in one patient. CONCLUSION: The described double staining technique could be a novel valuable tool that may help to achieve optimal anatomical and functional recovery after surgery for premacular fibrosis PMID- 12770968 TI - Visual performance after interface haemorrhage during laser in situ keratomileusis. AB - AIM: To study the visual performance in eyes with interface haemorrhage during laser assisted in situ keratomileusis (LASIK). METHODS: Case records of 20 patients, who had bleeding from the limbal vessels in one eye during LASIK (group 1) and uncomplicated surgery in the fellow eye (group 2) were studied. The parameters evaluated were uncorrected visual acuity (UCVA) best corrected visual acuity (BCVA), spherical equivalent of refraction (SEQ), contrast sensitivity, and glare acuity preoperatively and at 1, 3, and 6 months postoperatively. RESULTS: The mean preoperative SEQ in group 1 and 2 eyes was -5.79 (2.3) D and 5.27 (1.68) D, respectively. The mean decimal UCVA at 6 months after LASIK in group 1 and 2 eyes were 0.6 (0.2) and 1.0 respectively (p<0.001). The mean decimal BCVA at 1 week after LASIK in group 1 and 2 eyes were 0.89 (0.04) and 1.0 respectively (p<0.05). However, all eyes had a BCVA of 6/6 at 1, 3, and 6 months after LASIK. The mean contrast sensitivity values preoperatively in group 1 and 2 eyes were 161.3 (8.7) and 172 (68.2) respectively. There was a significant decrease in group 1 at 6 months (102 (60.5) (p<0.01)) compared to group 2. The decimal glare acuity preoperatively in group 1 and 2 eyes was 0.95 (0.11) and 0.89 (0.12), respectively. It decreased significantly in group 1 (0.7) (0.1 (p<0.01)) compared to group 2 at the 6 month follow up. CONCLUSION: Occurrence of intraoperative interface haemorrhage may affect the visual performance following LASIK surgery. PMID- 12770969 TI - Intraocular pressure and visual field loss in primary angle closure and primary open angle glaucomas. AB - AIM: To compare the correlation between visual field loss and the pretreatment intraocular pressure (IOP) in primary angle closure glaucoma (PACG) and primary open angle glaucoma (POAG). METHODS: In a cross sectional observational study of 74 patients (43 PACG, 31 POAG), pretreatment IOP was measured at presentation, before treatment was initiated. The severity of visual field loss was assessed by AGIS score, mean deviation (MD), pattern standard deviation (PSD), and corrected pattern standard deviation (CPSD). Glaucomatous optic neuropathy was assessed from simultaneous stereo disc photographs. RESULTS: There was a stronger correlation between pretreatment IOP and the extent of visual field loss in PACG subjects than in those with POAG for both MD (PACG: Pearson correlation coefficient (r) = 0.43, p = 0.002; r(2) = 0.19), (POAG: r = 0.21, p = 0.13; r(2) = 0.04) and AGIS score (PACG: r = 0.41, p = 0.003; r(2) = 0.17), (POAG: r = 0.23, p = 0.19; r(2) = 0.05 respectively). No such associations were seen for pattern standard deviation (PSD) or corrected pattern standard deviation (CPSD) in either group (p> 0.29). Both horizontal and vertical cup-disc ratio were well correlated with severity of field loss but not with presenting IOP for either diagnosis. CONCLUSIONS: This is consistent with the hypothesis of a greater IOP dependence for optic nerve damage in PACG than POAG and, conversely, a greater importance of other, less pressure dependent mechanisms in POAG compared to PACG. PMID- 12770970 TI - Interobserver agreement on visual field progression in glaucoma: a comparison of methods. AB - AIM: To examine the level of agreement between clinicians in assessing progressive deterioration in visual field series using two different methods of analysis. METHODS: Each visual field series satisfied the following criteria: more than 19 reliable fields, patient age over 40 years, macular threshold at least 30 dB. The first three fields in each series were excluded to minimise learning effects: the following 16 were studied. Five expert clinicians assessed the progression status of each series using both standard Humphrey printouts and pointwise linear regression (PROGRESSOR). The level of agreement between the clinicians was evaluated using a weighted kappa statistic. RESULTS: A total of 432 tests comprising 27 visual field series of 16 tests each were assessed by the clinicians. The level of agreement on progression status between the clinicians was always higher when they used PROGRESSOR (median kappa = 0.59) than when they used Humphrey printouts (median kappa = 0.32). This was statistically significant (p = 0.006, Wilcoxon matched pairs signed rank sum test). CONCLUSIONS: Agreement between expert clinicians about visual field progression status is poor when standard Humphrey printouts are used, even when the field series studied are long and consist solely of reliable fields. Under these ideal conditions, clinicians agree more closely about patients' visual field progression status when using PROGRESSOR than when inspecting series of Humphrey printouts. PMID- 12770972 TI - Epiretinal membrane removal in diabetic eyes: comparison of viscodissection with conventional methods of membrane peeling. AB - AIMS: To compare conventional methods of epiretinal membrane peeling with viscodissection. METHODS: 154 eyes with proliferative diabetic retinopathy (PDR) that underwent pars plana vitrectomy with membrane dissection (89 traditional, 65 viscodissection) were studied retrospectively. Incidence of retinal breaks (RBs), length of time under anaesthesia, postoperative intraocular pressure, retinal reattachment rate, and final visual acuity (VA) were measured. RESULTS: To compare cases of similar complexity, a "complexity score" was defined. The average complexity score for cases done with and without viscodissection was 4.7 and 3.2, respectively. The mean frequency of RBs in eyes undergoing viscodissection was 0.43 (SD 0.5) v 0.14 (0.35) RBs/eye without viscodissection. In complex cases, the frequency of posterior/peripheral RBs was 0.31 (0.47)/0.13 (0.34) RBs/eye, respectively, with viscodissection v 0.12 (0.33)/0.23 (0.43) RBs/eye without viscodissection. None of these differences were statistically significant. The average preoperative/postoperative VA (logMAR) in the viscodissection cohort was 1.7/1.3 (range 0.3 to >1.9/0.1 to >1.9) v 1.4/1 (range 0.48 to >1.9/0.1 to >1.9) in the non-viscodissection cohort, among eyes with 6 months of follow up. Anaesthesia duration was significantly shorter for cases done without viscodissection (p=0.03), but cases done with viscodissection were significantly more complex than cases done without viscodissection (p<0.0001). CONCLUSION: Viscodissection appears to be a safe and effective alternative technique in eyes with PDR. Owing to the retrospective nature of the study, additional studies are warranted. PMID- 12770971 TI - Colour Doppler imaging and fluorescein filling defects of the optic disc in normal tension glaucoma. AB - AIM: To investigate the relation between blood flow parameters of the retrobulbar vessels measured by means of colour Doppler imaging (CDI) and fluorescein filling defects of the optic nerve head in patients with normal tension glaucoma (NTG) and control subjects. METHODS: 29 patients with NTG and 29 age and sex matched control subjects were included in this study. Blood flow velocities-peak systolic velocity (PSV), end diastolic velocity (EDV), and resistive indices (RI) of the ophthalmic artery (OA), the central retinal artery (CRA), and of the temporal and nasal short posterior ciliary arteries (TPCA, NPCA)-were measured with CDI. Fluorescein angiograms were performed with a scanning laser ophthalmoscope. The extent of absolute fluorescein filling defects of the optic nerve head in relation to the optic nerve head was assessed. RESULTS: The PSV of the OA, the PSV and EDV of the CRA, and of the TPCA and NPCA were significantly reduced in NTG (p<0.05). The RI of the CRA, the TPCA and NPCA were significantly increased in NTG (p<0.01). The optic nerve head fluorescein filling defects were significantly larger in NTG (p<0.01). The filling defects were significantly negatively correlated (p<0.05) with the PSV and EDV of the CRA (PSV(CRA): r = 0.41; EDV(CRA): r = -0.34), with the PSV and EDV of the NPCA (PSV(NPCA): r = 0.34; EDV(NPCA): r = -0.38), and with the EDV of the TPCA (r = -0.29). A significant positive correlation (p<0.05) was found with the RI of both PCAs (RI(NPCA): r = 0.28; RI(TPCA): r = 0.29). CONCLUSION: Patients with NTG had reduced blood flow velocities and higher resistive indices in most retrobulbar vessels. Optic nerve head fluorescein filling defects were larger compared to controls. The filling defects were correlated with end diastolic velocities and resistive indices of the PCAs and with blood flow velocities of the CRA. Capillary loss of the optic nerve head may be related to higher downstream resistance and reduced blood flow velocities of the retrobulbar vessels. PMID- 12770973 TI - Extent of foveal tritanopia in diabetes mellitus. AB - AIM: To use a colour matching technique to test the hypothesis that the foveal tritanopic zone is increased in size in diabetes mellitus. METHOD: A Wright tristimulus colorimeter was adapted for small field colour matching and colour matches were performed on bipartite fields in the range 12' to 60' of arc. The reference stimulus was 490 nm desaturated with 650 nm and the matching stimulus consisted of either two wavelengths (530 nm and 650 nm) or three (460 nm, 530 nm, and 650 nm). The size of the zone of foveal tritanopia was measured using two alternative forced choice presentations of dichromatic and trichromatic matches made by the observer for different field sizes. 21 diabetic and 12 controls performed the experiment. RESULTS: The results for the controls show a normal distribution, with a median foveal tritanopic zone of 18' of arc. The median for the diabetic patients was also 18' of arc, but the distribution showed a significant skew to the right. A non-parametric test shows a significant difference in comparison with the controls (p = 0.01), with several subjects having extensive zones of foveal tritanopia, reaching up to 1 degree. CONCLUSIONS: In the majority of diabetic subjects the extent of foveal tritanopia is normal; however, there is good evidence that in a small number of subjects the size of the zone is significantly increased. This indicates S-cone pathway damage that is sufficiently severe to lead to dichromatic colour vision in the fovea. PMID- 12770975 TI - The effects of dorzolamide on choroidal and retinal perfusion in non-exudative age related macular degeneration. AB - AIM: To comprehensively evaluate the effects of dorzolamide on the choroidal and retinal circulation in patients with age related macular degeneration (AMD). METHODS: In this randomised, double masked, parallel study, 36 non-exudative AMD patients were randomised in a 2 to 1 fashion to placebo versus topical dorzolamide and underwent assessment of their choroidal and retinal circulation. Scanning laser ophthalmoscope indocyanine green angiograms (ICGA) were analysed by a new area dilution analysis technique. Four areas in the perifoveal region and two areas in the temporal peripapillary region were evaluated by plotting intensity of fluorescence of each area over time. The means of the choroidal filling times and the heterogeneity of the filling times were assessed. Scanning laser ophthalmoscope fluorescein angiography (FA) was evaluated for retinal arteriovenous passage (AVP) times by plotting intensity of fluorescence of retinal vessels over time. Assessment was performed at baseline and at 4 months. RESULTS: Compared to placebo, AMD patients treated with dorzolamide showed a significantly increased rapidity of choroidal filling in the superior and inferior peripapillary regions (p=0.007, p=0.02, respectively). No significant difference in choroidal filling times was found in any of the perifoveal areas (p=0.9). Also, on FA assessment, treatment with dorzolamide showed no statistical differences in AVP times (p=0.19). CONCLUSIONS: Dorzolamide may increase peripapillary choroidal perfusion in non-exudative AMD patients. Further studies are merited. PMID- 12770974 TI - Assessment of colour vision as a screening test for sight threatening diabetic retinopathy before loss of vision. AB - AIM: To assess the effects of sight threatening diabetic retinopathy (STDR) on colour vision and to evaluate automated tritan contrast threshold (TCT) testing for STDR screening before significant visual loss. METHOD: Patients were recruited from a hospital based photographic screening clinic. All subjects underwent best corrected Snellen visual acuity (BCVA) and those with 20/30 vision or worse were excluded. Automated TCT was performed with a computer controlled, cathode ray tube based technique. The system produced a series of sinusoidal, standardised equiluminant chromatic gratings along a tritan confusion axis. Grading of diabetic retinopathy was made by one of the team of experienced ophthalmic registrars (SpR) using slit lamp biomicroscopy and a 78D lens; HbA(1c) and urine albumin were also tested. RESULTS: Patients with STDR had significantly worse TCT despite normal BCVA (p<0.0001). TCT yielded a sensitivity of 100% for detecting diabetic maculopathy and 94% for STDR with a specificity of 95%. Logistic regression analyses showed that TCT (p<0.001) and HbA(1c) (p<0.05) correlated significantly with the presence of STDR but duration of diabetes, urine albumin counts, and BCVA failed to show any significant correlation. No associations between TCT and duration of disease, TCT and HbA(1c), and TCT and urine albumin counts were found. CONCLUSION: Tritan colour vision deficiency was observed in patients with STDR despite their normal BCVA. These results indicate that automated TCT assessment is an effective and clinically viable technique for detecting STDR, particularly diabetic maculopathy, before visual loss. PMID- 12770976 TI - Morphological and functional analyses of adult onset vitelliform macular dystrophy. AB - AIM: To evaluate the morphology and visual function of the macula in eyes with adult onset vitelliform macular dystrophy (AVMD). METHODS: 12 eyes of six patients with AVMD were examined by ophthalmoscopy, scanning laser ophthalmoscopy (SLO), optical coherence tomography (OCT), and multifocal electroretinography (mfERGs). The macular lesions were bilateral in all patients and varied from the typical vitelliform (five eyes), faded vitelliform changes with retinal pigment epithelium (RPE) atrophy (five eyes), and a normal fovea associated with small flecks around the macula (two eyes). RESULTS: SLO demonstrated small abnormal bright spots in the deep retina throughout the posterior retina in all cases. OCT showed a highly reflective fusiform thickened layer at the level of the RPE and choriocapillaris in patients with a submacular yellow vitelliform lesion. A well circumscribed, optically clear space was observed beneath the retinal layer in the macular lesions with RPE atrophy. The mfERGs were significantly reduced not only in the macular area but also in the outermost ring (20-30 degrees ) of the mfERGs. CONCLUSIONS: The submacular materials that accumulate within the RPE or subepithelial layers reported in previous histopathological studies of vitelliform lesions can be detected by OCT. In the macular lesions with RPE atrophy, the material may have disappeared leaving a subretinal or subepithelial optical clear space. These SLO and mfERG observations suggest that the morphological and functional abnormalities may not be localised just in the macular area but may be present throughout the posterior pole in eyes with AVMD. PMID- 12770977 TI - Is granuloma annulare related to intermediate uveitis with retinal vasculitis? AB - AIM: To report on eight patients with severe idiopathic intermediate uveitis (IU) and granuloma annulare (GA), a self limiting cutaneous condition of unknown aetiology. METHODS: Retrospective case series. Clinical ophthalmic and dermatological data were studied and fluorescein angiography and skin biopsies were reviewed. RESULTS: All patients with idiopathic IU had similar ocular features (eight with vitritis, seven with retinal vasculitis) and developed complications such as cystoid macular oedema (n=5), cataract (n=4), and glaucoma (n=3). Systemic diseases were not found, but a localised type of GA was observed in all. CONCLUSION: Seven out of eight patients with IU and GA developed severe retinal vasculitis. Further studies are needed for a better understanding of this association, a common pathogenesis, and its eventual clinical consequences. PMID- 12770978 TI - The clinical features of albinism and their correlation with visual evoked potentials. AB - AIM: To investigate the relation between the clinical and electrophysiological abnormalities of patients undergoing visual evoked potential investigation for albinism. METHODS: 40 subjects with a probable or possible clinical diagnosis of albinism underwent pattern appearance and/or flash visual evoked potential (VEP) examination. The VEP findings are correlated with the clinical features of albinism determined by clinical examination and orthoptic assessment. RESULTS: The majority of patients with clinical evidence of albinism showed a contralateral predominance in the VEPs. There was close correlation between the clinical signs of albinism and the degree of contralateral VEP predominance. This manifested as an interhemispheric latency asymmetry to monocular pattern appearance stimulation but amplitude asymmetry to flash stimulation. The strongest correlation for pattern appearance interhemispheric latency difference was with foveal hypoplasia (rho = 0.58; p = 0.0003) followed by nystagmus (rho = 0.48; p = 0.0027) and iris transillumination (rho = 0.33; p = 0.039). The VEP abnormalities were of greater magnitude in those patients with most features of albinism. Several patients with apparently mild disorders of ocular pigmentation had small but significantly abnormal VEP latency asymmetries. CONCLUSION: There is a strong association between the magnitude of the interhemispheric latency asymmetry of the pattern appearance VEP, and of amplitude asymmetry of the flash VEP, with the clinical signs of albinism. The data are consistent with a spectrum of abnormalities in albinism involving both clinical expression and electrophysiological misrouting, which is wider than previously recognised. PMID- 12770980 TI - Architecture of arachnoid trabeculae, pillars, and septa in the subarachnoid space of the human optic nerve: anatomy and clinical considerations. AB - AIMS: To describe the anatomy and the arrangement of the arachnoid trabeculae, pillars, and septa in the subarachnoid space of the human optic nerve and to consider their possible clinical relevance for cerebrospinal fluid dynamics and fluid pressure in the subarachnoid space of the human optic nerve. METHODS: Postmortem study with a total of 12 optic nerves harvested from nine subjects without ocular disease. All optic nerves used in this study were obtained no later than 7 hours after death, following qualified consent for necropsy. The study was performed with transmission (TEM) and scanning electron microscopy (SEM). RESULTS: The subarachnoid space of the human optic nerve contains a variety of trabeculae, septa, and stout pillars that are arranged between the arachnoid and the pia layers of the meninges of the nerve. They display a considerable numeric and structural variability depending on their location within the different portions of the optic nerve. In the bulbar segment (ampulla), adjacent to the globe, a dense and highly ramified meshwork of delicate trabeculae is arranged in a reticular fashion. Between the arachnoid trabeculae, interconnecting velum-like processes are observed. In the mid-orbital segment of the orbital portion, the subarachnoid space is subdivided, and can appear even loosely chambered by broad trabeculae and velum-like septa at some locations. In the intracanalicular segment additionally, few stout pillars and single round trabeculae are observed. CONCLUSION: The subarachnoid space of the human optic nerve is not a homogeneous and anatomically empty chamber filled with cerebrospinal fluid, but it contains a complex system of arachnoid trabeculae and septa that divide the subarachnoid space. The trabeculae, septa, and pillars, as well as their arrangement described in this study, may have a role in the cerebrospinal fluid dynamics between the subarachnoid space of the optic nerve and the chiasmal cistern and may contribute to the understanding of the pathophysiology of asymmetric and unilateral papilloedema. All the structures described are of such delicate character that they can not even be visualised with high resolution magnetic resonance imaging (MRI). PMID- 12770979 TI - Impact of smoking on the response to treatment of thyroid associated ophthalmopathy. AB - BACKGROUND: In patients with Graves' disease, smoking considerably increases the incidence and severity of thyroid associated ophthalmopathy (TAO). The authors sought to determine if smoking also influences the course of TAO during treatment, and the efficacy of therapy. METHODS: 41 smokers and 19 non-smokers with moderate untreated TAO were included in this prospective study. All patients were treated with steroids and, 6 weeks after the beginning of drug therapy, with orbital irradiation. Follow up was performed 1.5, 4.5, 7.5, and 12 months after the beginning of the study. Proptosis, clinical activity score (CAS), and motility were evaluated. The extent of smoking was derived from the concentration of the haemoglobin adduct N-2-hydroxyethylvaline (HEV), a parameter of long term smoking. RESULTS: There was no difference in the clinical manifestations of TAO between smokers and non-smokers at the beginning of treatment. However, CAS decreased (p<0.05) and motility improved (p<0.02) significantly faster and to a greater extent in non-smokers than smokers. Inverse correlations between the CAS decrease and the HEV levels observed 4.5 and 7.5 months after the beginning of treatment and between the improvement of motility and the HEV levels after 1.5, 4.5, and 7.5 months indicated a dose dependence. Mean HEV levels did not vary much during the follow up period and were significantly different in smokers (mean 5.4 (SD 2.7) micro g/l) and non-smokers (mean 1.8 (1.3) micro g/l; p<0.01). CONCLUSION: Smoking influences the course of TAO during treatment in a dose dependent manner. The response to treatment is delayed and considerably poorer in smokers. PMID- 12770981 TI - View 1: minimal segmental buckling without drainage. PMID- 12770983 TI - View 2: the case for primary vitrectomy. PMID- 12770984 TI - View 3: The case for pneumatic retinopexy. PMID- 12770985 TI - Debate overview. Repair of a primary rhegmatogenous retinal detachment. PMID- 12770986 TI - Long term survival of patient with invasive aspergillosis involving orbit, paranasal sinus, and central nervous system. PMID- 12770989 TI - Simultaneous presentation of choroidal melanoma in mother and daughter. PMID- 12770987 TI - The effect of topical glaucoma medications evaluated by perimetry. PMID- 12770988 TI - Delayed therapeutic success with endoscopic cyclophoto-coagulation in treating refractory post-penetrating keratoplasty glaucoma. PMID- 12770990 TI - Relative hypersensitivity in healthy eye by frequency doubling perimetry in patients with severely damaged contralateral eye. PMID- 12770991 TI - Giant neurosensory detachments associated with disciform lesions in neovascular age related macular degeneration. PMID- 12770992 TI - Solitary choroidal tuberculoma in a patient with chest wall tuberculosis. PMID- 12770993 TI - Successful photodynamic therapy for subretinal neovascularisation due to Sorsby's fundus dystrophy: 1 year follow up. PMID- 12770994 TI - Association of HLA type and Mooren's Ulcer in Chinese in Taiwan. PMID- 12770995 TI - Corneal endothelial deposits secondary to rifabutin prophylaxis for Mycobacterium avium complex bacteraemia. PMID- 12770996 TI - Corneal ectasia following deep lamellar keratoplasty. PMID- 12770997 TI - Opacification of SC60B-OUV lens implant following routine phacoemulsification surgery: case report and EM study. PMID- 12770998 TI - Isolated foveal retinoschisis as a cause of visual loss in young females. PMID- 12771000 TI - Region- and type-specific induction of matrix metalloproteinases in post myocardial infarction remodeling. AB - BACKGROUND: Induction of matrix metalloproteinases (MMPs) contributes to adverse remodeling after myocardial infarction (MI). Whether a region- and type-specific distribution of MMPs occurs within the post-MI myocardium remained unknown. METHODS AND RESULTS: Ten sheep were instrumented with a sonomicrometry array to measure dimensions in 7 distinct regions corresponding to the remote, transition, and MI regions. Eight sheep served as reference controls. The relative abundance of representative MMP types and the tissue inhibitors of the MMPs (TIMPs) was quantified by immunoblotting. Segment length increased from baseline in the remote (24.9+/-5.4%), transition (18.0+/-2.9%), and MI (53.8+/-11.0%) regions at 8 weeks after MI (P<0.05) and was greatest in the MI region (P<0.05). Region- and type-specific changes in MMPs occurred after MI. For example, MMP-1 and MMP-9 abundance was unchanged in the remote, fell to 3+/-2% in the transition, and was undetectable in the MI region (P<0.05). MMP-13, MMP-8, and MT1-MMP increased by >300% in the transition and MI regions (P<0.05). TIMP abundance decreased significantly in the transition region after MI and fell to undetectable levels within the MI region. CONCLUSIONS: The unique findings of this study were 2-fold. First, changes in regional geometry after MI were associated with changes in MMP levels. Second, a region-specific portfolio of MMPs was induced after MI and was accompanied by a decline in TIMP levels, indicative of a loss of MMP inhibitory control. Targeting the regional imbalance between specific MMPs and TIMPs within the post-MI myocardium holds therapeutic potential. PMID- 12771001 TI - Restoration of endothelial function by increasing high-density lipoprotein in subjects with isolated low high-density lipoprotein. AB - BACKGROUND: Loss-of-function mutations in the ATP-binding cassette (ABCA)-1 gene locus are the underlying cause for familial hypoalphalipoproteinemia, providing a human isolated low-HDL model. In these familial hypoalphalipoproteinemia subjects, we evaluated the impact of isolated low HDL on endothelial function and the vascular effects of an acute increase in HDL. METHODS AND RESULTS: In 9 ABCA1 heterozygotes and 9 control subjects, vascular function was assessed by venous occlusion plethysmography. Forearm blood flow responses to the endothelium dependent and -independent vasodilators serotonin (5HT) and sodium nitroprusside, respectively, and the inhibitor of nitric oxide synthase NG-monomethyl-l-arginine (L-NMMA) were measured. Dose-response curves were repeated after systemic infusion of apolipoprotein A-I/phosphatidylcholine (apoA-I/PC) disks. At baseline, ABCA1 heterozygotes had decreased HDL levels (0.4+/-0.2 mmol/L; P<0.05), and their forearm blood flow responses to both 5HT (maximum, 49.0+/ 10.4%) and L-NMMA (maximum, -22.8+/-22.9%) were blunted compared with control subjects (both P< or =0.005). Infusion of apoA-I/PC disks increased plasma HDL to 1.3+/-0.4 mmol/L in ABCA1 heterozygotes, which resulted in complete restoration of vasomotor responses to both 5HT and L-NMMA (both P90% reduction). A significant reduction in the expression of cyclin B1 protein (approximately 45%) was observed only in LNCaP cells. A 24 h exposure of PC-3 and LNCaP cells to an apoptosis-inducing concentration of AITC (20 micro M) resulted in a significant decrease (31-68%) in the levels of anti apoptotic protein Bcl-2 in both cell lines, and approximately 58% reduction in Bcl-X(L) protein expression in LNCaP cells. In conclusion, it seems reasonable to hypothesize that AITC, and possibly other ITCs, may find use in the treatment of human prostate cancers. PMID- 12771035 TI - Association of NAD(P)H: quinone oxidoreductase 1 (NQO1) C609T polymorphism with esophageal squamous cell carcinoma in a German Caucasian and a northern Chinese population. AB - NAD(P)H: quinone oxidoreductase 1 (NQO1) is an antioxidant enzyme, important in the detoxification of environmental carcinogens. A single base substitution (C - > T) polymorphism at nucleotide 609 (null-allele) of NQO1 gene impairs stability and function of the NQO1 protein. To investigate the association of this NQO1 polymorphism with susceptibility to esophageal squamous cell carcinoma (ESCC), the NQO1 C609T genotypes were determined by PCR-RFLP analysis in 450 patients with ESCC (257 German Caucasians and 193 northern Chinese) and 393 unrelated healthy controls (252 German Caucasians and 141 northern Chinese). Additionally, NQO1 protein expression was determined by immunohistochemistry in a subset of 74 ESCC (50 German, 24 Chinese). A significant difference in NQO1 C609T genotype distribution was observed between Caucasian healthy controls (C/C, 73.4%; C/T, 25.0%; T/T, 1.6%) and Chinese healthy controls (C/C, 34.0%; C/T, 49.7%; T/T, 16.3%) (chi(2) = 68.40, P < 0.001). The NQO1 T/T genotype significantly increased the risk for developing ESCC in both Caucasian subjects (OR = 4.62, 95% CI = 1.54 13.86) and Chinese subjects (OR = 1.81, 95% CI = 1.04-3.15), compared with the combined C/C and C/T genotypes. In Chinese subjects, this increased susceptibility was pronounced in patients with family history of upper gastrointestinal cancers (OR = 2.18, 95% CI = 1.14-4.17). Immunohistochemical analysis showed NQO1 protein expression in 53 carcinomas, whereas 21 carcinomas were negative. Negativity for NQO1 expression correlated strongly with the NQO1 genotype, being present in 8.6% of cases with C/C, 22.2% of cases with C/T and 100% of cases with T/T genotype (chi(2) = 16.60, P < 0.001). In summary, the association of the NQO1 C609T polymorphism with ESCC in genetically distinct populations makes a strong argument for its importance in carcinogenesis of ESCC in the German Caucasian and the northern Chinese population. PMID- 12771036 TI - Mutagenic events induced by 4-hydroxyequilin in supF shuttle vector plasmid propagated in human cells. AB - Increased incidence of breast, ovarian and endometrial cancers are observed in women receiving estrogen replacement therapy (ERT). Equilin and equilenin are the major components of the widely prescribed drug used for ERT. These equine estrogens are metabolized primarily to 4-hydroxyequilin (4-OHEQ) and 4 hydroxyequilenin, respectively, which are autoxidized to react with DNA, resulting in the various DNA damages. To explore the mutagenic potential of equine estrogen metabolites, a double-stranded pMY189 shuttle vector carrying a bacteria suppressor tRNA gene, supF, was exposed to 4-OHEQ and transfected into human fibroblast. Plasmids containing mutations in the supF gene were detected with indicator bacteria and mutated colonies obtained were analyzed by automatic DNA sequencing. The proportion of plasmids with the mutated supF gene was increased dose-dependently. The majority of the 4-OHEQ-induced mutations were base substitutions (78%); another 22% were deletions and insertions. Among the base substitutions, 56% were single base substitutions and 19% were multiple base substitutions. The majority (86%) of the 4-OHEQ-induced single base substitutions occurred at the C:G site. C:G --> G:C and C:G --> A:T mutations were detected preferentially with lesser numbers of C:G --> T:A transitions. Sixty-two percent of base substitutions were observed particularly at C:G pairs in (5')-TC/AG-(5') sequences. Using (32)P-post-labeling/gel electrophoresis analysis, 4-OHEN-dC was a major adduct, followed by lesser amounts of 4-OHEN-dA adduct. Mutations observed at C:G pairs may result from 4-OHEN-dC adduct. These results indicated that 4-OHEQ is mutagenic, generating mutations primarily at C:G pairs in (5') TC/AG-(5') sequences. PMID- 12771037 TI - Effect of eicosapentaenoic acid, an omega-3 polyunsaturated fatty acid, on UVR related cancer risk in humans. An assessment of early genotoxic markers. AB - Dietary omega-3 polyunsaturated fatty acids (omega-3 PUFAs) protect against photocarcinogenesis in animals, but prospective human studies are scarce. The mechanism(s) underlying the photoprotection are uncertain, although omega-3 PUFAs may influence oxidative stress. We examined the effect of supplementation on a range of indicators of ultraviolet radiation (UVR)-induced DNA damage in humans, and assessed effect on basal and post-UVR oxidative status. In a double-blind randomized study, 42 healthy subjects took 4 g daily of purified omega-3 PUFA, eicosapentaenoic acid (EPA), or monounsaturated, oleic acid (OA), for 3 months. EPA was bioavailable; the skin content at 3 months showing an 8-fold rise from baseline, P < 0.01. No consistent pattern of alteration in basal and UVR-exposed skin content of the antioxidants glutathione, vitamins E and C or lipid peroxidation, was seen on supplementation. Sunburn sensitivity was reduced on EPA, the UVR-induced erythemal threshold rising from a mean of 36 (SD 10) mJ/cm(2) at baseline to 49 (16) mJ/cm(2) after supplementation, P < 0.01. Moreover, UVR-induced skin p53 expression, assessed immunohistochemically at 24 h post-UVR exposure, fell from a mean of 16 (SD 5) positive cells/100 epidermal cells at baseline to 8 (4) after EPA supplementation, P < 0.01. Peripheral blood lymphocytes (PBL) sampled on 3 successive days both pre- and post supplementation, showed no change with respect to basal DNA single-strand breaks or oxidative base modification (8-oxo-dG). However, when susceptibility of PBL to ex vivo UVR was examined using the comet assay, this revealed a reduction in tail moment from 84.4 (SD 3.4) at baseline to 69.4 (3.1) after EPA, P = 0.03. No significant changes were seen in any of the above parameters following OA supplementation. Reduction in this range of early markers, i.e. sunburn, UVR induced p53 in skin and strand breaks in PBL, indicate protection by dietary EPA against acute UVR-induced genotoxicity; longer-term supplementation might reduce skin cancer in humans. PMID- 12771038 TI - Treatment of green tea polyphenols in hydrophilic cream prevents UVB-induced oxidation of lipids and proteins, depletion of antioxidant enzymes and phosphorylation of MAPK proteins in SKH-1 hairless mouse skin. AB - The use of botanical supplements has received immense interest in recent years to protect human skin from adverse biological effects of solar ultraviolet (UV) radiation. The polyphenols from green tea are one of them and have been shown to prevent photocarcinogenesis in animal models but their mechanism of photoprotection is not well understood. To determine the mechanism of photoprotection in in vivo mouse model, topical treatment of polyphenols from green tea (GTP) or its most chemopreventive constituent (-)-epigallocatechin-3 gallate (EGCG) (1 mg/cm(2) skin area) in hydrophilic ointment USP before single (180 mJ/cm(2)) or multiple UVB exposures (180 mJ/cm(2), daily for 10 days) resulted in significant prevention of UVB-induced depletion of antioxidant enzymes such as glutathione peroxidase (78-100%, P < 0.005-0.001), catalase (51 92%, P < 0.001) and glutathione level (87-100%, P < 0.005). Treatment of EGCG or GTP also inhibited UVB-induced oxidative stress when measured in terms of lipid peroxidation (76-95%, P < 0.001), and protein oxidation (67-75%, P > 0.001). Further, to delineate the inhibition of UVB-induced oxidative stress with cell signaling pathways, treatment of EGCG to mouse skin resulted in marked inhibition of a single UVB irradiation-induced phosphorylation of ERK1/2 (16-95%), JNK (46 100%) and p38 (100%) proteins of MAPK family in a time-dependent manner. Identical photoprotective effects of EGCG or GTP were also observed against multiple UVB irradiation-induced phosphorylation of the proteins of MAPK family in vivo mouse skin. Photoprotective efficacy of GTP given in drinking water (d.w.) (0.2%, w/v) was also determined and compared with that of topical treatment of EGCG and GTP. Treatment of GTP in d.w. also significantly prevented single or multiple UVB irradiation-induced depletion of antioxidant enzymes (44 61%, P < 0.01-0.001), oxidative stress (33-71%, P < 0.01) and phosphorylation of ERK1/2, JNK and p38 proteins of MAPK family but the photoprotective efficacy was comparatively less than that of topical treatments of EGCG and GTP. Lesser photoprotective efficacy of GTP in d.w. in comparison with topical application may be due to its less bioavailability in skin target cells. Together, for the first time a cream based formulation of green tea polyphenols was tested in this study to explore the possibility of its use for the humans, and the data obtained from this in vivo study further suggest that GTP could be useful in attenuation of solar UVB light-induced oxidative stress-mediated and MAPK-caused skin disorders in humans. PMID- 12771039 TI - Dietary folate deficiency suppresses N-methyl-N-nitrosourea-induced mammary tumorigenesis in rats. AB - Epidemiologic studies have suggested that dietary folate intake is inversely related to breast cancer risk. However, epidemiologic evidence has not been consistent nor has it provided unequivocal support for this purported inverse relationship. This study investigated the effect of dietary folate on N-methyl-N nitrosourea (MNU)-induced mammary tumorigenesis in rats. Weanling, female Sprague Dawley rats were fed diets containing either 0 (deficient; n = 22), 2 (basal dietary requirement, control; n = 20) or 8 mg (supplemented; n = 20) folate/kg diet for 30 weeks. At 50 days of age, rats received an i.p. injection of MNU (50 mg/kg body wt). At necropsy, all macroscopic mammary tumors were identified and examined microscopically. The effect of dietary folate on genomic DNA methylation in mammary tumorigenesis was determined by the in vitro methyl acceptance assay. The incidence of mammary adenoma and adenocarcinoma in the folate-deficient group was lower than that of the control and folate-supplemented groups (55 versus 90 and 75%, respectively, P = 0.043). Kaplan-Meier analyses also demonstrated a similar trend in the rates of appearance of either adenoma or adenocarcinoma (P = 0.06). In contrast, folate supplementation did not significantly modulate mammary tumorigenesis compared with the control group. Although mammary tumors were significantly hypomethylated compared with non-neoplastic mammary tissues in each dietary group (P < 0.03), folate status did not significantly affect the extent of DNA methylation. The data suggest that dietary folate deficiency of a moderate degree suppresses, whereas folate supplementation at four times the basal dietary requirement does not significantly modulate, mammary tumorigenesis in this model. The role of folate in mammary tumorigenesis needs to be clarified for safe and effective prevention of breast cancer. PMID- 12771040 TI - Celecoxib and difluoromethylornithine in combination have strong therapeutic activity against UV-induced skin tumors in mice. AB - The cyclooxygenase-2 (COX-2) inhibitor celecoxib and the ornithine decarboxylase (ODC) inhibitor difluoromethylornithine (DFMO) were each previously shown to prevent skin tumor development when administered throughout the course of UV irradiation. This raised the question of whether maintenance or continued growth of existing tumors required prostaglandins, the product of COX, or polyamines, the product of ODC. To address this question, SKH hairless mice were irradiated 3 times/week with 90 mJ/cm(2); this dose was increased 10% weekly to a maximum of 175 mJ/cm(2). UV was stopped at 27 weeks, at which time there were an average of 5 papillomas/mouse. The mice were then placed in one of four treatment groups: group 1, no treatment; group 2, 0.4% DFMO in the drinking water; group 3, 500 p.p.m. celecoxib in the diet (AIN76); group 4, both DFMO and celecoxib. The control group continued to produce new tumors in a nearly linear manner such that by week 31 the tumor number had nearly doubled, i.e. approximately 10 tumors/mouse. The group receiving DFMO showed significant tumor regression, losing an average of 1 tumor/mouse/week, such that 50% of the tumors remained at week 31. The celecoxib group showed a 25% reduction in tumor number. The group receiving the combination of celecoxib and DFMO showed the greatest regression, with an 89% reduction in tumor number compared with the control group. There was also a corresponding reduction in the size of the tumors. To determine whether tumor regression was permanent or required continued treatment, all treatments were stopped at 31 weeks. Over the next 4 weeks, tumors reappeared at the same rate in all treatment groups. It is concluded that the combination of celecoxib and DFMO are potent therapeutic agents for skin cancer, although the benefits are lost with the cessation of treatment. PMID- 12771042 TI - Induction of oxidative DNA damage by arsenite and its trivalent and pentavalent methylated metabolites in cultured human cells and isolated DNA. AB - Even though a well-known human carcinogen the underlying mechanisms of arsenic carcinogenicity are still not fully understood. For arsenite, proposed mechanisms are the interference with DNA repair processes and an increase in reactive oxygen species. Even less is known about the genotoxic potentials of its methylated metabolites monomethylarsonous [MMA(III)] and dimethylarsinous [DMA(III)] acid, monomethylarsonic [MMA(V)] and dimethylarsinic [DMA(V)] acid. Within the present study we compared the induction of oxidative DNA damage by arsenite and its methylated metabolites in cultured human cells and in isolated PM2 DNA, by frequencies of DNA strand breaks and of lesions recognized by the bacterial formamidopyrimidine-DNA glycosylase (Fpg). Only DMA(III) (> or =10 micro M) generated DNA strand breaks in isolated PM2 DNA. In HeLa S3 cells, short-term incubations (0.5-3 h) with doses as low as 10 nM arsenite induced high frequencies of Fpg-sensitive sites, whereas the induction of oxidative DNA damage after 18 h incubation was rather low. With respect to the methylated metabolites, both trivalent and pentavalent metabolites showed a pronounced induction of Fpg sensitive sites in the nanomolar or micromolar concentration range, respectively, which was present after both short-term and long-term incubations. Furthermore MMA(III) and DMA(V) generated DNA strand breaks in a concentration-dependent manner. Taken together our results show that very low physiologically relevant doses of arsenite and the methylated metabolites induce high levels of oxidative DNA damage in cultured human cells. Thus, biomethylation of inorganic arsenic may be involved in inorganic arsenic-induced genotoxicity/carcinogenicity. PMID- 12771041 TI - Immortal, telomerase-negative cell lines derived from a Li-Fraumeni syndrome patient exhibit telomere length variability and chromosomal and minisatellite instabilities. AB - Five immortal cell lines derived from a Li-Fraumeni syndrome patient (MDAH 087) with a germline mutant p53 allele were characterized with respect to telomere length and genomic instability. The remaining wild-type p53 allele is lost in the cell lines. Telomerase activity was undetectable in all immortal cell lines. Five subclones of each cell line and five re-subclones of each of the subclones also showed undetectable telomerase activity. All five immortal cell lines exhibited variability in the mean length of terminal restriction fragments (TRFs). Subclones of each cell line, and re-subclones of the subclones also showed TRF variability, indicating that the variability is owing to clonal heterogeneity. Chromosome aberrations were observed at high frequencies in these cell lines including the subclones and re-subclones, and the principal types of aberrations were breaks, double minute chromosomes and dicentric chromosomes. In addition, minisatellite instability detected by DNA fingerprints was observed in the immortal cell lines. However, all of the cell lines were negative for microsatellite instability. As minisatellite sequences are considered recombinogenic in mammalian cells, these results suggest that recombination rates can be increased in these cell lines. Tumor-derived human cell lines, HT1080 cells and HeLa cells that also lack p53 function, exhibited little genomic instability involving chromosomal and minisatellite instabilities, indicating that chromosomal and minisatellite instabilities observed in the immortal cell lines lacking telomerase activity could not result from loss of p53 function. PMID- 12771043 TI - Molecular profiling of hepatocellular carcinomas developing spontaneously in acyl CoA oxidase deficient mice: comparison with liver tumors induced in wild-type mice by a peroxisome proliferator and a genotoxic carcinogen. AB - By using cDNA microarrays, we studied the expression profiles of 26 hepatocellular carcinomas (HCC) developing spontaneously in peroxisomal fatty acyl-CoA oxidase null (AOX-/-) mice. The development of liver tumors in AOX-/- mice is due to sustained activation of peroxisome proliferator-activated receptor alpha (PPARalpha) by the unmetabolized substrates of AOX, which serve as natural PPARalpha ligands. We then compared the AOX-/- liver tumor expression profiles with those induced by ciprofibrate, a non-genotoxic peroxisome proliferator, or by the genotoxic carcinogen diethylnitrosamine (DENA) to discern differences in gene expression patterns that may predict or distinguish PPARalpha-mediated liver tumors from genotoxically derived tumors. Our results show that HCCs developing in AOX-/- mice share a number of deregulated (up- or down-regulated) genes with ciprofibrate-induced liver tumors. The overall commonality of expression between AOX-/- and ciprofibrate-induced liver tumors but not with DENA-induced tumors strongly implicates the activation of PPARalpha and PPARalpha-regulated genes in liver, including those participating in lipid catabolism, as key factors in the development of HCC in AOX-/- and in ciprofibrate-treated mice. Northern blot analysis confirmed the differential expression of some of the genes identified in the present study, and also some genes identified previously as PPARalpha regulated, such as CD36, lymphocyte antigen 6 complex locus (Ly-6D), and C3f. We found a panel of 12 genes upregulated in all three classes of liver tumors, namely AOX-/-, ciprofibrate-induced and DENA-induced. These include an uncharacterized RIKEN cDNA, lipocalin 2, insulin-like growth factor-binding protein 1, Ly-6D and CD63 among others. In conclusion, these results identify distinguishing features between non-genotoxic and genotoxic carcinogen derived liver tumors as well as genes that are upregulated in both types and suggest that RIKEN cDNA, Ly-6D and lipocalin 2 in particular appear to be desirable molecular markers for further study in liver carcinogenesis and progression. PMID- 12771044 TI - Enhancement of colon carcinogenesis by prostaglandin E2 administration. AB - Although an accumulating body of evidence indicates that levels of prostaglandin E(2) (PGE(2)) in human and rodent colon cancers are higher than those in surrounding normal tissues, the precise contribution of PGE(2) to the process of colon cancer development has still been unclear. Therefore, we designed a study using a well-established azoxymethane (AOM)-induced colon carcinogenesis in male F344 rat model to investigate whether administration of exogenous PGE(2) has a real impact on colon carcinogenesis. Intraperitoneal PGE(2) injections (7.7 micro g) once a week for 25 weeks significantly increased the AOM-induced colon tumor incidence (percent rats with tumors, 92 versus 53%, P < 0.05), especially adenocarcinomas (92 versus 47%, P < 0.05), and multiplicity (number of tumors per rat, 2.8 versus 1.0, P < 0.05). PGE(2) treatment significantly increased 5-bromo 2'-deoxyuridine (BrdUrd) labeling index (11.8 versus 9.7%, P < 0.05) and reduced apoptotic index (0.34 versus 0.53%, P < 0.05) in colon cancers induced by AOM. PGE(2) exhibits its physiological functions through binding to E-prostanoid (EP) membrane receptors EP(1-4). All four types of EP receptors were detected in AOM induced colon cancers using reverse transcription-polymerase chain reaction (RT PCR). Our results provide evidence that PGE(2) enhances colon carcinogenesis through induction of cell proliferation and reduction of apoptosis. PMID- 12771045 TI - Predictors of the plasma ratio of 2-hydroxyestrone to 16alpha-hydroxyestrone among pre-menopausal, nulliparous women from four ethnic groups. AB - Studies of circulating estrogen levels in relation to pre-menopausal breast cancer risk have yielded inconsistent results. Various estrogen metabolites might affect the risk differently. Estradiol metabolism occurs primarily via two mutually exclusive pathways, yielding 2-hydroxyestrone (2-OHE) and 16alpha hydroxyestrone (16alpha-OHE). Most, but not all, studies have found that a relatively high 2-OHE/16alpha-OHE ratio is associated with a low breast cancer risk. Our objective was to determine if the 2-OHE/16alpha-OHE ratio in plasma correlates with suspected breast cancer risk factors and other lifestyle factors, such as ethnicity, body size, age at menarche, oral contraceptive use, smoking, vegetarian diet, coffee and alcohol consumption in 513 nulliparous women, aged 17 35. Oral contraceptive users had significantly lower 2-OHE/16alpha-OHE ratios than pill non-users (P = 10(-21)). Among women who were not using oral contraceptives, the median 2-OHE/16alpha-OHE ratio in plasma was similar for white, black, Indian/Pakistani and Asian women, after adjustment for age and menstrual cycle phase. Among oral contraceptive users, Asian women had significantly lower 2-OHE/16alpha-OHE ratios than white women, and this result remained after adjustment for age and day of menstrual cycle. Daily coffee consumption was significantly positively correlated with 2-OHE/16alpha-OHE ratios (r(s) = 0.18, P = 0.002) only among pill non-users. Our findings suggest that the plasma 2-OHE/16alpha-OHE ratio is associated with constitutional factors and with modifiable lifestyle factors. The reported elevated risk of early onset breast cancer among young oral contraceptive users could be mediated in part through altered estrogen metabolism induced by synthetic estrogens and progestins. PMID- 12771046 TI - Methylation of 2-hydroxyestradiol in isolated organs. AB - Vascular smooth muscle and glomerular mesangial cells in culture express a biochemical pathway that methylates 2-hydroxyestradiol (17beta-estradiol metabolite) to produce 2-methoxyestradiol, a cell growth inhibitor that may mediate the cardiorenal protective effects of 17beta-estradiol. Whether this pathway exists in intact organ systems is currently unclear. Accordingly, the purpose of the present investigation was to characterize the methylation of 2 hydroxestradiol in intact organs from both male and female rats. No significant differences were detected in the ability of male and female tissues to methylate 2-hydroxyestradiol. In isolated hearts, kidneys, and mesenteries perfused with Tyrode's solution, Km values for 2-hydroxyestradiol methylation were 0.175+/ 0.021, 0.387+/-0.054, and 0.495+/-0.089 micromol/L, respectively, and Vmax values were 21.0+/-1.58, 24.9+/-1.49, and 1.01+/-0.148 pmol 2-methoxyestradiol x min(-1) x ml(-1) per gram, respectively. The catalytic efficiency (Vmax/Km) was greatest in the heart compared with the kidney and mesentery (132+/-14.3, 78.4+/-15.1, and 2.30+/-0.263 pmol 2-methoxyestradiol x min(-1) x mL(-1) x micromol/L(-1) per gram, respectively). In the kidney, the catechol-O-methyltransferase inhibitor quercetin and norepinephrine (10 micromol/L) reduced methylation of 2 hydroxyestradiol by approximately 90% and 41%, respectively. Importantly, methylation in the kidney was inhibited by an average of 16.6+/-1.80% by endogenous norepinephrine released by renal artery nerve stimulation. Our results indicate that a robust 2-hydroxyestradiol methylation pathway exists in the kidney and heart, but not in the mesentery, and that this pathway is mediated by catechol-O-methyltransferase. Our findings also suggest that catecholamines may interfere with 2-hydroxyestradiol methylation and thereby attenuate the cardiorenal protective effects of 17beta-estradiol. PMID- 12771048 TI - GLUT-1 overexpression: Link between hemodynamic and metabolic factors in glomerular injury? AB - Mesangial matrix deposition is the hallmark of hypertensive and diabetic glomerulopathy. At similar levels of systemic hypertension, Dahl salt-sensitive but not spontaneously hypertensive rats (SHR) develop glomerular hypertension, which is accompanied by upregulation of transforming growth factor beta1 (TGF beta1), mesangial matrix expansion, and sclerosis. GLUT-1 is ubiquitously expressed and is the predominant glucose transporter in mesangial cells. In mesangial cells in vitro, GLUT-1 overexpression increases basal glucose transport, resulting in excess fibronectin and collagen production. TGF-beta1 has been shown to upregulate GLUT-1 expression. We demonstrated that in hypertensive Dahl salt-sensitive (S) rats fed 4% NaCl (systolic blood pressure [SBP]: 236+/-9 mm Hg), but not in similarly hypertensive SHR (SBP: 230+/-10 mm Hg) or their normotensive counterparts (Dahl S fed 0.5% NaCl, SBP: 145+/-5 mm Hg; and Wistar Kyoto, SBP: 137+/-3 mm Hg), there was an 80% upregulation of glomerular GLUT-1 protein expression (P< or =0.03). This was accompanied by a 2.7-fold upregulation of TGF-beta1 protein expression in glomeruli of DSH compared with DSN rats (P=0.02). TGF-beta1 expression was not upregulated and did not differ in the glomeruli of Wistar-Kyoto and SHR rats. As an in vitro surrogate of the in vivo hemodynamic stress imposed by glomerular hypertension, we used mechanical stretching of human and rat mesangial cells. We found that after 33 hours of stretching, mesangial cells overexpressed GLUT-1 (40%) and showed an increase in basal glucose transport of similar magnitude (both P< or =0.01), which could be blocked with an anti TGF-beta1-neutralizing antibody. These studies suggest a novel link between hemodynamic and metabolic factors that may cooperate in inducing progressive glomerular injury in conditions characterized by glomerular hypertension. PMID- 12771047 TI - Imbalance of T-cell subsets in angiotensin II-infused hypertensive rats with kidney injury. AB - Blockade of angiotensin (Ang) II is efficient in various renal diseases. Although interest has focused on the hemodynamic changes and reduction of proteinuria, recent studies emphasize the nonhemodynamic effects of Ang II on kidney injury. The aim of this study was to clarify the mechanisms of Ang II on the immune system that alter the balance of helper T-cell (Th) subsets. We used a continuous, Ang II infusion model of rats that develop hypertension, proteinuria, and tubulointerstitial damage, including de novo expression of alpha-smooth muscle actin and loss of endothelial cells. We isolated T cells from the spleen and measured cytokine levels by ELISA systems. Ang II-infused rats showed an increase in the Th1 cytokine gamma-interferon and a decrease in the Th2 cytokine interleukin-4. The same change in cytokine mRNA expression in the spleen and kidney was confirmed by quantitative polymerase chain reaction analysis. Our ELISPOT assay showed an increase in the number of gamma-interferon-secreting T cells by Ang II. To investigate whether these changes were specific effects of Ang II, we treated the model rats with the Ang II receptor blocker (ARB) olmesartan or the nonspecific vessel dilator hydralazine. Administration of the ARB ameliorated disease manifestations and the imbalance in Th subsets, whereas hydralazine did not, despite comparable effects on blood pressure. These results demonstrate a direct role of Ang II in the modification of Th balance. The imbalance of Th subsets was associated with hypertensive kidney injury induced by Ang II. Some of the beneficial effects of ARBs might be explained by their immunomodulatory reactions. PMID- 12771049 TI - Sustained activation of the central baroreceptor pathway in obesity hypertension. AB - The major goal of this study was to determine whether there is increased activation of medullary neurons that participate in the central baroreceptor reflex pathway in dogs with obesity-induced hypertension, a model of hypertension that is associated with increased sympathetic activity. We used Fos-like (Fos-Li) protein immunohistochemical methods to determine activation of neurons in the nucleus tractus solitarius (NTS), caudal ventrolateral medulla (CVLM), and rostral ventrolateral medulla (RVLM). Dogs were fed either a regular diet or an identical diet with the addition of 0.5 to 0.9 kg of cooked beef fat. After approximately 6 weeks of the high fat diet, body weight (36.3+/-0.4 vs 21.5+/-0.5 kg), mean arterial pressure (105+/-4 vs 91+/-3 mm Hg), and heart rate (97+/-4 vs 70+/-3 bpm) were significantly greater in obese than in control dogs, respectively. There was little Fos-Li immunoreactivity in medullary neurons of control dogs but marked reactivity in obese dogs. Specifically, the number of Fos Li-positive cells in the NTS and CVLM was 3 to 5 times greater in obese than in control dogs. Furthermore, despite sustained activation of these baroreceptor sensitive neurons, there was a significantly greater number of Fos-Li positive cells in the RVLM of dogs fed the high fat diet. As baroreceptor suppression of sympathoexcitatory cells in the RVLM is mediated by activation of neurons in the NTS and CVLM, these results support recent findings indicating that baroreflex suppression of sympathetic activity is a long-term compensatory response in hypertension. However, sympathoexcitatory inputs onto RVLM neurons would appear to predominate over the inhibitory effects of the baroreflex in obesity hypertension. PMID- 12771050 TI - Lessons from SARS. PMID- 12771053 TI - Pertussis control in Canada. PMID- 12771054 TI - Physicians for oral health. PMID- 12771055 TI - Enlightening medical students. PMID- 12771056 TI - Enlightening medical students. PMID- 12771057 TI - Safe use of acetaminophen. PMID- 12771059 TI - ECT for Parkinson's? PMID- 12771060 TI - Transmission of HPV. PMID- 12771062 TI - The return of "negative" trials? PMID- 12771064 TI - QALYs: the best option so far. PMID- 12771065 TI - First-use risks. PMID- 12771066 TI - Why choose ophthalmology? PMID- 12771068 TI - West Nile virus infection in 2002: morbidity and mortality among patients admitted to hospital in southcentral Ontario. AB - BACKGROUND: In August and September 2002 an outbreak of West Nile virus (WNV) infection occurred in southern Ontario. We encountered a number of seriously ill patients at our hospitals. In this article we document the clinical characteristics of these cases. METHODS: We conducted a retrospective chart review of patients who came to the attention of infectious disease or neurology consultants or the microbiology laboratories at 7 hospitals in the municipalities of Toronto, Peel and Halton, Ont. Patients were included if they had been admitted to hospital or stayed overnight in the emergency department, had serological evidence of WNV infection and had clinical evidence of WNV fever, aseptic meningitis, encephalomyelitis or motor neuronopathy. RESULTS: In all, 64 patients met the inclusion criteria; 57 had encephalitis or neuromuscular weakness or both, 5 had aseptic meningitis, and 2 had WNV fever. The mean age was 61 years (range 26-87). The patients were predominantly active, middle-aged or elderly people living independently in the community. Seven patients were immunocompromised A febrile prodromal illness preceded the neurological symptoms in almost all cases. The most common neurological abnormality was decreased level of consciousness; this frequently evolved to severe lower motor neuron neuromuscular weakness. Ataxia and swallowing disorders were frequent and important problems. Sixteen patients (25%) required intubation and mechanical ventilation because of a decreased level of consciousness, inability to clear secretions or respiratory muscle weakness; 9 others had disabling muscle weakness of one or more limbs. Ten patients died. The study patients were in hospital a total of 1856 patient-days, including 532 patient-days in an intensive care unit. Only 28% (13/47) of the patients who survived encephalitis or neuromuscular weakness, or both, were discharged home without additional support. Slow turnaround time for serological test results resulted in delayed diagnosis. INTERPRETATION: The 2002 WNV infection outbreak in Ontario caused serious morbidity and mortality in the subset of patients who had encephalitis or neuromuscular weakness severe enough to require hospital admission. PMID- 12771070 TI - Identification and containment of an outbreak of SARS in a community hospital. AB - BACKGROUND: Severe acute respiratory syndrome (SARS) is continuing to spread around the world. All hospitals must be prepared to care for patients with SARS. Thus, it is important to understand the transmission of this disease in hospitals and to evaluate methods for its containment in health care institutions. We describe how we cared for the first 2 patients with SARS admitted to our 419-bed community hospital in Richmond Hill, Ont., and the response to a SARS outbreak within our institution. METHODS: We collected clinical and epidemiological data about patients and health care workers at our institution who during a 13-day period had a potential unprotected exposure to 2 patients whose signs and symptoms were subsequently identified as meeting the case definition for probable SARS. The index case at our hospital was a patient who was transferred to our intensive care unit (ICU) from a referral hospital on Mar. 16, 2003, where he had been in close proximity to the son of the individual with the first reported case of SARS in Toronto. After 13 days in the ICU, a diagnosis of probable SARS was reached for our index case. Immediately upon diagnosis of our index case, respiratory isolation and barrier precautions were instituted throughout our hospital and maintained for a period of 10 days, which is the estimated maximum incubation period reported for this disease. Aggressive surveillance measures among hospital staff, patients and visitors were also maintained during this time. RESULTS: During the surveillance period, 15 individuals (10 hospital staff, 3 patients and 2 visitors) were identified as meeting the case definition for probable or suspected SARS, in addition to our index case. All but 1 individual had had direct contact with a symptomatic patient with SARS during the period of unprotected exposure. No additional cases were identified after infection control precautions had been implemented for 8 days. No cases of secondary transmission were identified in the 21 days following the implementation of these precautions at our institution. INTERPRETATION: SARS can easily be spread by direct personal contact in the hospital setting. We found that the implementation of aggressive infection control measures is effective in preventing further transmission of this disease. PMID- 12771069 TI - Differences in operative mortality between high- and low-volume hospitals in Ontario for 5 major surgical procedures: estimating the number of lives potentially saved through regionalization. AB - BACKGROUND: Previous research has shown that persons undergoing certain high-risk surgical procedures at high-volume hospitals (HVHs) have a lower risk of postoperative death than those undergoing surgery at low-volume hospitals (LVHs). We estimated the absolute number of operative deaths that could potentially be avoided if 5 major surgical procedures in Ontario were restricted to HVHs. METHODS: We collected data on all persons who underwent esophagectomy (613), colon or rectal resection for colorectal cancer (18 898), pancreaticoduodenectomy (686), pulmonary lobectomy or pneumonectomy for lung cancer (5156) or repair of an unruptured abdominal aortic aneurysm (AAA) (6279) in Ontario from Apr. 1, 1994, to Mar. 31, 1999. We calculated the excess number of operative deaths (defined as deaths in the period from the day of the operation to 30 days thereafter), adjusted for age, sex and comorbidity, among the 75% of persons treated in LVHs, as compared with the 25% treated in the highest-volume quartile of hospitals. Bootstrap methods were used to estimate 95% confidence intervals (CIs). RESULTS: Of the 31 632 persons undergoing any of the 5 procedures, 1341 (4.24%) died within 30 days of surgery. If the 75% of persons treated at the LVHs had instead been treated at the HVHs, the annual number of lives potentially saved would have been 4 (95% CI, 0 to 9) for esophagectomy, 6 (95% CI, 1 to 11) for pancreaticoduodenectomy, 1 (95% CI, -10 to 13) for major lung resection and 14 (95% CI, 1 to 25) for repair of unruptured AAA. For resection of colon or rectum, the regionalization strategy would not have saved any lives, and 17 lives (95% CI, 36 to -3) would potentially have been lost. INTERPRETATION: A small number of operative deaths are potentially avoidable by performing 4 of 5 complex surgical procedures only at HVHs in Ontario. In determining health policy, the most compelling argument for regionalizing complex surgical procedures at HVHs may not be the prevention of a large number of such deaths. PMID- 12771071 TI - Enteroviruses and sudden deafness. AB - A young, healthy man presented with sudden severe sensorineural hearing loss and tinnitus. The results of the workup and neuroimaging were normal, as were the auditory brain stem responses. Methylprednisolone pulse therapy was associated with significant hearing improvement within 10 days. A history of a short self limited febrile illness preceding admission (with headache, photophobia, myalgia and fatigue), a raised serum C-reactive protein level and transient leukopenia suggested an infectious cause. Lumbar puncture revealed a mononuclear pleocytosis of the cerebrospinal fluid, with negative cultures but positive polymerase chain reaction test results for enterovirus, which was later cultured from the patient's stool. The patient's wife and baby had had a similar febrile illness without hearing loss 10 days earlier, and an outbreak of enterovirus meningitis was identified in the area, which was associated with familial clustering and echovirus serotype 4 infection. The varied causes of sudden sensorineural hearing loss, which should include enterovirus, are reviewed here. PMID- 12771072 TI - Pesticide use for West Nile virus. PMID- 12771073 TI - Excluding pulmonary embolism with helical (spiral) computed tomography: Evidence is catching up with enthusiasm. PMID- 12771074 TI - SARS: prudence, not panic. PMID- 12771075 TI - Why was Toronto included in the World Health Organization's SARS-related travel advisory? PMID- 12771076 TI - Serotonin syndrome: a brief review. PMID- 12771077 TI - West Nile virus. PMID- 12771078 TI - Nonnarcotic analgesic use and the risk of hypertension. PMID- 12771079 TI - The eyes have it: conjugate eye deviation on CT scan aids in early detection of ischemic stroke. PMID- 12771083 TI - Larvicide debate marks start of another West Nile virus summer. PMID- 12771080 TI - Concerns over lindane treatment for scabies and lice. PMID- 12771089 TI - SARS poses challenges for MDs treating pediatric patients. PMID- 12771091 TI - SARS may have a silver lining, WHO says. PMID- 12771095 TI - A piece of my mind. What I have seen. PMID- 12771093 TI - Burden of home care borne by women. PMID- 12771096 TI - Detecting acute HIV infections feasible, North Carolina program demonstrates. PMID- 12771097 TI - Naval hospital ship heads for home after treating casualties of Gulf war. PMID- 12771098 TI - From the Centers for Disease Control and Prevention. Updated interim surveillance case definition for severe acute respiratory syndrome (SARS)--United States, April 29, 2003. PMID- 12771099 TI - From the Centers for Disease Control and Prevention. Update: severe acute respiratory syndrome--United States, 2003. PMID- 12771100 TI - From the Centers for Disease Control and Prevention. Self-reported asthma prevalence and control among adults--United States, 2001. PMID- 12771101 TI - From the Centers for Disease Control and Prevention. Poisoning by an illegally imported chinese rodenticide containing tetramethylenedisulfotetramine--New York City, 2002. PMID- 12771102 TI - From the Centers for Disease Control and Prevention. Update: adverse events following civilian smallpox vaccination--United States, 2003. PMID- 12771103 TI - Optimal digoxin concentrations for patients with heart failure. PMID- 12771104 TI - Optimal digoxin concentrations for patients with heart failure. PMID- 12771105 TI - Health and function of patients with untreated idiopathic scoliosis. PMID- 12771106 TI - Approaches to reducing consumption of alcohol by youth. PMID- 12771107 TI - Liability for adverse events in direct-to-consumer advertising. PMID- 12771108 TI - Liability for adverse events in direct-to-consumer advertising. PMID- 12771109 TI - Measuring improvement in quality of care. PMID- 12771110 TI - Can administrative data assess physicians' quality of care? PMID- 12771111 TI - Analysis of generic nevirapine products in developing countries. PMID- 12771112 TI - Estrogen plus progestin and the incidence of dementia and mild cognitive impairment in postmenopausal women: the Women's Health Initiative Memory Study: a randomized controlled trial. AB - CONTEXT: Postmenopausal women have a greater risk than men of developing Alzheimer disease, but studies of the effects of estrogen therapy on Alzheimer disease have been inconsistent. On July 8, 2002, the study drugs, estrogen plus progestin, in the Women's Health Initiative (WHI) trial were discontinued because of certain increased health risks in women receiving combined hormone therapy. OBJECTIVE: To evaluate the effect of estrogen plus progestin on the incidence of dementia and mild cognitive impairment compared with placebo. DESIGN, SETTING, AND PARTICIPANTS: The Women's Health Initiative Memory Study (WHIMS), a randomized, double-blind, placebo-controlled clinical trial, began enrolling participants from the Women's Health Initiative (WHI) estrogen plus progestin trial in May 1996. Of the 4894 eligible participants of the WHI study, 4532 (92.6%) postmenopausal women free of probable dementia, aged 65 years or older, and recruited from 39 of 40 WHI clinical centers were enrolled in the WHIMS. INTERVENTION: Participants received either 1 daily tablet of 0.625 mg of conjugated equine estrogen plus 2.5 mg of medroxyprogesterone acetate (n = 2229), or a matching placebo (n = 2303). MAIN OUTCOME MEASURES: Incidence of probable dementia (primary outcome) and mild cognitive impairment (secondary outcome) were identified through a structured clinical assessment. RESULTS: The mean (SD) time between the date of randomization into WHI and the last Modified Mini-Mental State Examination (3MSE) for all WHIMS participants was 4.05 (1.19) years. Overall, 61 women were diagnosed with probable dementia, 40 (66%) in the estrogen plus progestin group compared with 21 (34%) in the placebo group. The hazard ratio (HR) for probable dementia was 2.05 (95% confidence interval [CI], 1.21 3.48; 45 vs 22 per 10 000 person-years; P =.01). This increased risk would result in an additional 23 cases of dementia per 10 000 women per year. Alzheimer disease was the most common classification of dementia in both study groups. Treatment effects on mild cognitive impairment did not differ between groups (HR, 1.07; 95% CI, 0.74-1.55; 63 vs 59 cases per 10 000 person-years; P =.72). CONCLUSIONS: Estrogen plus progestin therapy increased the risk for probable dementia in postmenopausal women aged 65 years or older. In addition, estrogen plus progestin therapy did not prevent mild cognitive impairment in these women. These findings, coupled with previously reported WHI data, support the conclusion that the risks of estrogen plus progestin outweigh the benefits. PMID- 12771113 TI - Effect of estrogen plus progestin on global cognitive function in postmenopausal women: the Women's Health Initiative Memory Study: a randomized controlled trial. AB - CONTEXT: Observational studies have suggested that postmenopausal hormone treatment may improve cognitive function, but data from randomized clinical trials have been sparse and inconclusive. The Women's Health Initiative Memory Study (WHIMS) is an ancillary study of the Women's Health Initiative (WHI) hormone therapy trials. On July 8, 2002, the estrogen plus progestin therapy in the WHI trial was discontinued because of certain increased health risks for women. OBJECTIVE: To determine whether estrogen plus progestin therapy protects global cognitive function in older postmenopausal women. DESIGN, SETTING, AND PARTICIPANTS: A randomized, double-blind, placebo-controlled clinical trial, WHIMS is an ancillary study of geographically diverse, community-dwelling women aged 65 years or older from 39 of 40 clinical centers within the WHI estrogen plus progestin trial that started in June 1995. Of 4894 eligible postmenopausal women aged 65 years or older and free of probable dementia at baseline, 4532 (92.6%) were enrolled in the estrogen plus progestin component of WHIMS. A total of 4381 participants (96.7%) provided at least 1 valid cognitive function score between June 1995 and July 8, 2002. INTERVENTIONS: Participants received either 1 daily tablet containing 0.625 mg of conjugated equine estrogen with 2.5 mg of medroxyprogesterone acetate (n = 2145) or matching placebo (n = 2236). MAIN OUTCOME MEASURE: Global cognitive function measured annually with the Modified Mini-Mental State Examination. RESULTS: The Modified Mini-Mental State Examination mean total scores in both groups increased slightly over time (mean follow-up of 4.2 years). Women in the estrogen plus progestin group had smaller average increases in total scores compared with women receiving placebo (P =.03), but these differences were not clinically important. Removing women by censoring them after adjudicated dementia, mild cognitive impairment, or stroke, and nonadherence to study protocol, did not alter the findings. Prior hormone therapy use and duration of prior use did not affect the interpretation of the results, nor did timing of prior hormone therapy initiation with respect to the final menstrual period. More women in the estrogen plus progestin group had a substantial and clinically important decline (> or =2 SDs) in Modified Mini Mental State Examination total score (6.7%) compared with the placebo group (4.8%) (P =.008). CONCLUSIONS: Among postmenopausal women aged 65 years or older, estrogen plus progestin did not improve cognitive function when compared with placebo. While most women receiving estrogen plus progestin did not experience clinically relevant adverse effects on cognition compared with placebo, a small increased risk of clinically meaningful cognitive decline occurred in the estrogen plus progestin group. PMID- 12771114 TI - Effect of estrogen plus progestin on stroke in postmenopausal women: the Women's Health Initiative: a randomized trial. AB - CONTEXT: The Women's Health Initiative (WHI) trial of estrogen plus progestin was stopped early because of adverse effects, including an increased risk of stroke in the estrogen plus progestin group. OBJECTIVE: To assess the effect of estrogen plus progestin on ischemic and hemorrhagic stroke and in subgroups, and to determine whether the effect of estrogen plus progestin was modified by baseline levels of blood biomarkers. DESIGN: Multicenter double-blind, placebo-controlled, randomized clinical trial involving 16 608 women aged 50 through 79 years with an average follow-up of 5.6 years. Baseline levels of blood-based markers of inflammation, thrombosis, and lipid levels were measured in the first 140 centrally confirmed stroke cases and 513 controls. INTERVENTIONS: Participants received 0.625 mg/d of conjugated equine estrogen plus 2.5 mg/d of medroxyprogesterone acetate (n = 8506) or placebo (n = 8102). MAIN OUTCOME MEASURES: Overall strokes and stroke subtype and severity were centrally adjudicated by stroke neurologists. RESULTS: One hundred fifty-one patients (1.8%) in the estrogen plus progestin and 107 (1.3%) in the placebo groups had strokes. Overall 79.8% of strokes were ischemic. For combined ischemic and hemorrhagic strokes, the intention-to-treat hazard ratio (HR) for estrogen plus progestin vs placebo was 1.31 (95% confidence interval [CI], 1.02-1.68); with adjustment for adherence, the HR was 1.50 (95% CI, 1.08-2.08). The HR for ischemic stroke was 1.44 (95% CI, 1.09-1.90) and for hemorrhagic stroke, 0.82 (95% CI, 0.43-1.56). Point estimates of the HRs indicate that excess risk of all stroke was apparent in all age groups, in all categories of baseline stroke risk, and in women with and without hypertension, prior history of cardiovascular disease, use of hormones, statins, or aspirin. Other risk factors for stroke, including smoking, blood pressure, diabetes, lower use of vitamin C supplements, blood-based biomarkers of inflammation, higher white blood cell count, and higher hematocrit levels did not modify the effect of estrogen plus progestin on stroke risk. CONCLUSIONS: Estrogen plus progestin increases the risk of ischemic stroke in generally healthy postmenopausal women. Excess risk for all strokes attributed to estrogen plus progestin appeared to be present in all subgroups of women examined. PMID- 12771115 TI - Combined cardiac resynchronization and implantable cardioversion defibrillation in advanced chronic heart failure: the MIRACLE ICD Trial. AB - CONTEXT: Cardiac resynchronization therapy (CRT) through biventricular pacing is an effective treatment for heart failure (HF) with a wide QRS; however, the outcomes of patients requiring CRT and implantable cardioverter defibrillator (ICD) therapy are unknown. OBJECTIVE: To examine the efficacy and safety of combined CRT and ICD therapy in patients with New York Heart Association (NYHA) class III or IV congestive HF despite appropriate medical management. DESIGN, SETTING, AND PARTICIPANTS: Randomized, double-blind, parallel-controlled trial conducted from October 1, 1999, to August 31, 2001, of 369 patients with left ventricular ejection fraction of 35% or less, QRS duration of 130 ms, at high risk of life-threatening ventricular arrhythmias, and in NYHA class III (n = 328) or IV (n = 41) despite optimized medical treatment. INTERVENTIONS: Of 369 randomized patients who received devices with combined CRT and ICD capabilities, 182 were controls (ICD activated, CRT off) and 187 were in the CRT group (ICD activated, CRT on). MAIN OUTCOME MEASURES: The primary double-blind study end points were changes between baseline and 6 months in quality of life, functional class, and distance covered during a 6-minute walk. Additional outcome measures included changes in exercise capacity, plasma neurohormones, left ventricular function, and overall HF status. Survival, incidence of ventricular arrhythmias, and rates of hospitalization were also compared. RESULTS: At 6 months, patients assigned to CRT had a greater improvement in median (95% confidence interval) quality of life score (-17.5 [-21 to -14] vs -11.0 [-16 to -7], P =.02) and functional class (-1 [-1 to -1] vs 0 [-1 to 0], P =.007) than controls but were no different in the change in distance walked in 6 minutes (55 m [44-79] vs 53 m [43-75], P =.36). Peak oxygen consumption increased by 1.1 mL/kg per minute (0.7 1.6) in the CRT group vs 0.1 mL/kg per minute (-0.1 to 0.8) in controls (P =.04), although treadmill exercise duration increased by 56 seconds (30-79) in the CRT group and decreased by 11 seconds (-55 to 12) in controls (P<.001). No significant differences were observed in changes in left ventricular size or function, overall HF status, survival, and rates of hospitalization. No proarrhythmia was observed and arrhythmia termination capabilities were not impaired. CONCLUSIONS: Cardiac resynchronization improved quality of life, functional status, and exercise capacity in patients with moderate to severe HF, a wide QRS interval, and life-threatening arrhythmias. These improvements occurred in the context of underlying appropriate medical management without proarrhythmia or compromised ICD function. PMID- 12771116 TI - Variation of serum prostate-specific antigen levels: an evaluation of year-to year fluctuations. AB - CONTEXT: Serum prostate-specific antigen (PSA) testing is frequently used in early detection programs for prostate cancer. While PSA testing has resulted in an increase in prostate cancer detection, its routine use has been questioned because of a lack of specificity. OBJECTIVE: To determine whether year-to-year fluctuations in PSA levels are due to natural variation and render a single PSA test result unreliable. DESIGN, SETTING, AND PARTICIPANTS: Retrospective analysis of an unscreened population of 972 men (median age, 62 years) participating in the Polyp Prevention Trial (1991-1998). Five consecutive blood samples were obtained during a 4-year period and were assessed for total and free PSA levels. MAIN OUTCOME MEASURE: Abnormal PSA test result based on a PSA level higher than 4 ng/mL; a PSA level higher than 2.5 ng/mL; a PSA level above the age-specific cutoff; a PSA level in the range of 4 to 10 ng/mL and a free-to-total ratio of less than 0.25 ng/mL; or a PSA velocity higher than 0.75 ng/mL per year. RESULTS: Prostate biopsy would have been recommended in 207 participants (21%) with a PSA level higher than 4 ng/mL; in 358 (37%) with a level higher than 2.5 ng/mL; in 172 (18%) with a level above the age-specific cutoff; in 190 (20%) with a level between 4 and 10 ng/mL and a free-to-total ratio of less than 0.25 ng/mL; and in 145 (15%) with a velocity higher than 0.75 ng/mL per year. Among men with an abnormal PSA finding, a high proportion had a normal PSA finding at 1 or more subsequent visits during 4-year follow-up: 68 (44%) of 154 participants with a PSA level higher than 4 ng/mL; 116 (40%) of 291 had a level higher than 2.5 ng/mL; 64 (55%) of 117 had an elevated level above the age-specific cutoff; and 76 (53%) of 143 had a level between 4 and 10 ng/mL and a free-to-total ratio of less than 0.25 ng/mL. CONCLUSION: An isolated elevation in PSA level should be confirmed several weeks later before proceeding with further testing, including prostate biopsy. PMID- 12771117 TI - A 65-year-old woman with acute cough illness and an important engagement. PMID- 12771118 TI - Use of race and ethnicity in biomedical publication. AB - Researchers, clinicians, and policy makers face 3 challenges in writing about race and ethnicity: accounting for the limitations of race/ethnicity data; distinguishing between race/ethnicity as a risk factor or as a risk marker; and finding a way to write about race/ethnicity that does not stigmatize and does not imply a we/they dichotomy between health professionals and populations of color. Journals play an important role in setting standards for research and policy literature. The authors outline guidelines that might be used when race and ethnicity are addressed in biomedical publications. PMID- 12771119 TI - Hormone therapy and the brain: deja vu all over again? PMID- 12771120 TI - Combining resynchronization and defibrillation therapies for heart failure. PMID- 12771121 TI - JAMA patient page. Coughs, colds, and antibiotics. PMID- 12771122 TI - Is HP1 an RNA detector that functions both in repression and activation? AB - Heterochromatin is defined as regions of compact chromatin that persist throughout the cell cycle (Heitz, 1928). The earliest cytological observations of heterochromatin were followed by ribonucleotide labeling experiments that showed it to be transcriptionally inert relative to the more typical euchromatic regions that decondense during interphase. Genetic studies of rearrangements that place euchromatic genes next to blocks of heterochromatin also pointed out the repressive nature of heterochromatin (Grigliatti, 1991; and references therein). The discovery of the heterochromatin-enriched protein heterochromatin protein 1 (HP1)**Abbreviation used in this paper: HP1, heterochromatin protein 1. by Elgin and co-workers in the mid-1980s suggested that the distinct cytological features of this chromatin may be related to its unique nucleoprotein composition (James and Elgin, 1986; James et al., 1989). HP1 immunostaining on polytene chromosomes from Drosophila larval salivary glands was used to show enrichment of the protein in pericentric heterochromatin. Since that initial discovery, HP1 homologues have been found in species ranging from fission yeast to humans where it is associated with gene silencing (Eissenberg and Elgin, 2000; and references therein). A number of euchromatic sites of localization were also reported in this original study. It has been generally assumed that these sites might constitute euchromatic sites of transcriptional repression by HP1. Indeed, several genes located at one of these sites (cytological region 31) have increased transcript levels in mutants for HP1 (Hwang et al., 2001). PMID- 12771123 TI - Caveolae/raft-dependent endocytosis. AB - Although caveolae are well-characterized subdomains of glycolipid rafts, their distinctive morphology and association with caveolins has led to their internalization being considered different from that of rafts. In this review, we propose that caveolae and rafts are internalized via a common pathway, caveolae/raft-dependent endocytosis, defined by its clathrin independence, dynamin dependence, and sensitivity to cholesterol depletion. The regulatory role of caveolin-1 and ligand sorting in this complex endocytic pathway are specifically addressed. PMID- 12771124 TI - Different cofactor activities in gamma-secretase assembly: evidence for a nicastrin-Aph-1 subcomplex. AB - The gamma-secretase complex is required for intramembrane cleavage of several integral membrane proteins, including the Notch receptor, where it generates an active signaling fragment. Four putative gamma-secretase components have been identified-presenilin (Psn), nicastrin (Nct), Aph-1, and Pen-2. Here, we use a stepwise coexpression approach to investigate the role of each new component in gamma-secretase assembly and activation. Coexpression of all four proteins leads to high level accumulation of mature Psn and increased proteolysis of Notch. Aph 1 and Nct may form a subcomplex that stabilizes the Psn holoprotein at an early step in gamma-secretase assembly. Subcomplex levels of Aph-1 are down-regulated by stepwise addition of Psn, suggesting that Aph-1 might not enter the mature complex. In contrast, Pen-2 accumulates proportionally with Psn, and is associated with Psn endoproteolysis during gamma-secretase assembly. These results demonstrate that Aph-1 and Pen-2 are essential cofactors for Psn, but that they play different roles in gamma-secretase assembly and activation. PMID- 12771125 TI - Keratin 8 protection of placental barrier function. AB - The intermediate filament protein keratin 8 (K8) is critical for the development of most mouse embryos beyond midgestation. We find that 68% of K8-/- embryos, in a sensitive genetic background, are rescued from placental bleeding and subsequent death by cellular complementation with wild-type tetraploid extraembryonic cells. This indicates that the primary defect responsible for K8-/ lethality is trophoblast giant cell layer failure. Furthermore, the genetic absence of maternal but not paternal TNF doubles the number of viable K8-/- embryos. Finally, we show that K8-/- concepti are more sensitive to a TNF dependent epithelial apoptosis induced by the administration of concanavalin A (ConA) to pregnant mothers. The ConA-induced failure of the trophoblast giant cell barrier results in hematoma formation between the trophoblast giant cell layer and the embryonic yolk sac in a phenocopy of dying K8-deficient concepti in a sensitive genetic background. We conclude the lethality of K8-/- embryos is due to a TNF-sensitive failure of trophoblast giant cell barrier function. The keratin-dependent protection of trophoblast giant cells from a maternal TNF dependent apoptotic challenge may be a key function of simple epithelial keratins. PMID- 12771126 TI - Dishevelled activates Ca2+ flux, PKC, and CamKII in vertebrate embryos. AB - Wnt ligands and Frizzled (Fz) receptors have been shown to activate multiple intracellular signaling pathways. Activation of the Wnt-beta-catenin pathway has been described in greatest detail, but it has been reported that Wnts and Fzs also activate vertebrate planar cell polarity (PCP) and Wnt-Ca2+ pathways. Although the intracellular protein Dishevelled (Dsh) plays a dual role in both the Wnt-beta-catenin and the PCP pathways, its potential involvement in the Wnt Ca2+ pathway has not been investigated. Here we show that a Dsh deletion construct, XDshDeltaDIX, which is sufficient for activation of the PCP pathway, is also sufficient for activation of three effectors of the Wnt-Ca2+ pathway: Ca2+ flux, PKC, and calcium/calmodulin-dependent protein kinase II (CamKII). Furthermore, we find that interfering with endogenous Dsh function reduces the activation of PKC by Xfz7 and interferes with normal heart development. These data suggest that the Wnt-Ca2+ pathway utilizes Dsh, thereby implicating Dsh as a component of all reported Fz signaling pathways. PMID- 12771127 TI - Kinetic analysis of receptor-activated phosphoinositide turnover. AB - We studied the bradykinin-induced changes in phosphoinositide composition of N1E 115 neuroblastoma cells using a combination of biochemistry, microscope imaging, and mathematical modeling. Phosphatidylinositol-4,5-bisphosphate (PIP2) decreased over the first 30 s, and then recovered over the following 2-3 min. However, the rate and amount of inositol-1,4,5-trisphosphate (InsP3) production were much greater than the rate or amount of PIP2 decline. A mathematical model of phosphoinositide turnover based on this data predicted that PIP2 synthesis is also stimulated by bradykinin, causing an early transient increase in its concentration. This was subsequently confirmed experimentally. Then, we used single-cell microscopy to further examine phosphoinositide turnover by following the translocation of the pleckstrin homology domain of PLCdelta1 fused to green fluorescent protein (PH-GFP). The observed time course could be simulated by incorporating binding of PIP2 and InsP3 to PH-GFP into the model that had been used to analyze the biochemistry. Furthermore, this analysis could help to resolve a controversy over whether the translocation of PH-GFP from membrane to cytosol is due to a decrease in PIP2 on the membrane or an increase in InsP3 in cytosol; by computationally clamping the concentrations of each of these compounds, the model shows how both contribute to the dynamics of probe translocation. PMID- 12771128 TI - Contact inhibition of VEGF-induced proliferation requires vascular endothelial cadherin, beta-catenin, and the phosphatase DEP-1/CD148. AB - Confluent endothelial cells respond poorly to the proliferative signals of VEGF. Comparing isogenic endothelial cells differing for vascular endothelial cadherin (VE-cadherin) expression only, we found that the presence of this protein attenuates VEGF-induced VEGF receptor (VEGFR) 2 phosphorylation in tyrosine, p44/p42 MAP kinase phosphorylation, and cell proliferation. VE-cadherin truncated in beta-catenin but not p120 binding domain is unable to associate with VEGFR-2 and to induce its inactivation. beta-Catenin-null endothelial cells are not contact inhibited by VE-cadherin and are still responsive to VEGF, indicating that this protein is required to restrain growth factor signaling. A dominant negative mutant of high cell density-enhanced PTP 1 (DEP-1)//CD148 as well as reduction of its expression by RNA interference partially restore VEGFR-2 phosphorylation and MAP kinase activation. Overall the data indicate that VE cadherin-beta-catenin complex participates in contact inhibition of VEGF signaling. Upon stimulation with VEGF, VEGFR-2 associates with the complex and concentrates at cell-cell contacts, where it may be inactivated by junctional phosphatases such as DEP-1. In sparse cells or in VE-cadherin-null cells, this phenomenon cannot occur and the receptor is fully activated by the growth factor. PMID- 12771129 TI - Functional studies and distribution define a family of transmembrane AMPA receptor regulatory proteins. AB - Functional expression of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors in cerebellar granule cells requires stargazin, a member of a large family of four-pass transmembrane proteins. Here, we define a family of transmembrane AMPA receptor regulatory proteins (TARPs), which comprise stargazin, gamma-3, gamma-4, and gamma-8, but not related proteins, that mediate surface expression of AMPA receptors. TARPs exhibit discrete and complementary patterns of expression in both neurons and glia in the developing and mature central nervous system. In brain regions that express multiple isoforms, such as cerebral cortex, TARP-AMPA receptor complexes are strictly segregated, suggesting distinct roles for TARP isoforms. TARPs interact with AMPA receptors at the postsynaptic density, and surface expression of mature AMPA receptors requires a TARP. These studies indicate a general role for TARPs in controlling synaptic AMPA receptors throughout the central nervous system. PMID- 12771130 TI - Alpha(v)beta3 integrin expression up-regulates cdc2, which modulates cell migration. AB - The alphavbeta3 integrin has been shown to promote cell migration through activation of intracellular signaling pathways. We describe here a novel pathway that modulates cell migration and that is activated by alphavbeta3 and, as downstream effector, by cdc2 (cdk1). We report that alphavbeta3 expression in LNCaP (beta3-LNCaP) prostate cancer cells causes increased cdc2 mRNA levels as evaluated by gene expression analysis, and increased cdc2 protein and kinase activity levels. We provide three lines of evidence that increased levels of cdc2 contribute to a motile phenotype on integrin ligands in different cell types. First, increased levels of cdc2 correlate with more motile phenotypes of cancer cells. Second, ectopic expression of cdc2 increases cell migration, whereas expression of dominant-negative cdc2 inhibits migration. Third, cdc2 inhibitors reduce cell migration without affecting cell adhesion. We also show that cdc2 increases cell migration via specific association with cyclin B2, and we unravel a novel pathway of cell motility that involves, downstream of cdc2, caldesmon. cdc2 and caldesmon are shown here to localize in membrane ruffles in motile cells. These results show that cdc2 is a downstream effector of the alphavbeta3 integrin, and that it promotes cell migration. PMID- 12771131 TI - Repressors of androgen and progesterone receptor action. AB - Androgen and progesterone receptors (AR and PR) are two determining factors in gonadal differentiation that are highly expressed in developing and mature gonads. Loss of AR results in XY sex reversal and mutations causing reduced AR activity lead to varying degrees of defects in masculinization. Female PR knockout mice are infertile due to ovarian defects. While much has been discovered about positive regulation of these receptors by coactivators little is known about repression of the transcriptional activity of AR and PR in the presence of agonists. In this study we assessed the effect of SMRT and DAX-1 on AR and PR activity in the presence of both agonists and partial antagonists. We show that SMRT and DAX-1 repress agonist-dependent activity of both receptors, and the mechanism of repression includes disruption of the receptor dimer interactions rather than recruitment of histone deacetylases. We demonstrate that endogenous agonist-bound PR and DAX-1 in T47D breast cancer cells and endogenous AR and DAX-1 in LNCaP prostate cancer cells can be coimmunoprecipitated suggesting that the interaction is physiological. Surprisingly, although DAX-1 represses partial antagonist activity of AR, it was ineffective in repressing partial antagonist induced activity of PR. In contrast to most reported repressors, the expression of DAX-1 is restricted. We found that although DAX-1 is expressed in normal human prostate, its expression is strongly reduced in benign prostatic hyperplasia suggesting that DAX-1 plays a role in limiting AR activity in prostate. PMID- 12771132 TI - Adenomatous polyposis coli (APC)-independent regulation of beta-catenin degradation via a retinoid X receptor-mediated pathway. AB - Beta-catenin is a component of stable cell adherent complexes whereas its free form functions as a transcription factor that regulate genes involved in oncogenesis and metastasis. Free beta-catenin is eliminated by two adenomatous polyposis coli (APC)-dependent proteasomal degradation pathways regulated by glycogen synthase kinase 3beta (GSK3 beta) or p53-inducible Siah-1. Dysregulation of beta-catenin turnover consequent to mutations in critical genes of the APC dependent pathways is implicated in cancers such as colorectal cancer. We have identified a novel retinoid X receptor (RXR)-mediated APC-independent pathway in the regulation of beta-catenin. In this proteasomal pathway, RXR agonists induce degradation of beta-catenin and RXR alpha and repress beta-catenin-mediated transcription. In vivo, beta-catenin interacts with RXR alpha in the absence of ligand, but RXR agonists enhanced the interaction. RXR agonist action was not impaired by GSK3 beta inhibitors or deletion of the GSK3 beta-targeted sequence from beta-catenin. In APC- and p53-mutated colorectal cancer cells, RXR agonists still inactivated endogenous beta-catenin via RXR alpha. Interestingly, deletion of the RXR alpha A/B region abolished ligand-induced beta-catenin degradation but not RXR alpha-mediated transactivation. RXR alpha-mediated inactivation of oncogenic beta-catenin paralleled a reduction in cell proliferation. These results suggest a potential role for RXR and its agonists in the regulation of beta-catenin turnover and related biological events. PMID- 12771133 TI - Dynamic recruitment of NF-Y and histone acetyltransferases on cell-cycle promoters. AB - Regulation of transcription during the cell-cycle is under the control of E2 factors (E2Fs), often in cooperation with nuclear factor Y (NF-Y), a histone-like CCAAT-binding trimer. NF-Y is paradigmatic of a constitutive, ubiquitous factor that pre-sets the promoter architecture for other regulatory proteins to access it. We analyzed the recruitment of NF-Y, E2F1/4/6, histone acetyltransferases, and histone deacetylase (HDAC) 1/3/4 to several cell-cycle promoters by chromatin immunoprecipitation assays in serum-starved and restimulated NIH3T3 cells. NF-Y binding is not constitutive but timely regulated in all promoters tested, being displaced when promoters are repressed. p300 association correlates with activation, and it is never found in the absence of NF-Y, whereas PCAF/hGCN5 is often found before NF-Y association. E2F4 and E2F6, together with HDACs, are bound to repressed promoters, including the G2/M Cyclin B2. As expected, an inverse relationship between HDACs association and histones H3/H4 acetylation is observed. Blocking cells in G1 with the cyclin-dependent kinase 2 inhibitor R roscovitine confirms that NF-Y is bound to G1/S but not to G2/M promoters in G1. These data indicate that following the release of E2Fs/HDACs, a hierarchy of PCAF NF-Y-p300 interactions and H3-H4 acetylations are required for activation of cell cycle promoters. PMID- 12771134 TI - Dopamine prevents nitration of tyrosine hydroxylase by peroxynitrite and nitrogen dioxide: is nitrotyrosine formation an early step in dopamine neuronal damage? AB - Peroxynitrite and nitrogen dioxide (NO2) are reactive nitrogen species that have been implicated as causal factors in neurodegenerative conditions. Peroxynitrite induced nitration of tyrosine residues in tyrosine hydroxylase (TH) may even be one of the earliest biochemical events associated with 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine-induced damage to dopamine neurons. Exposure of TH to peroxynitrite or NO2 results in nitration of tyrosine residues and modification of cysteines in the enzyme as well as inactivation of catalytic activity. Dopamine (DA), its precursor 3,4-dihydroxyphenylalanine, and metabolite 3,4 dihydroxyphenylacetic acid completely block the nitrating effects of peroxynitrite and NO2 on TH but do not relieve the enzyme from inhibition. o Quinones formed in the reaction of catechols with either peroxynitrite or NO2 react with cysteine residues in TH and inhibit catalytic function. Using direct, real-time evaluation of tyrosine nitration with a green fluorescent protein-TH fusion protein stably expressed in intact cells (also stably expressing the human DA transporter), DA was also found to prevent NO2-induced nitration while leaving TH activity inhibited. These results show that peroxynitrite and NO2 react with DA to form quinones at the expense of tyrosine nitration. Endogenous DA may therefore play an important role in determining how DA neurons are affected by reactive nitrogen species by shifting the balance of their effects away from tyrosine nitration and toward o-quinone formation. PMID- 12771135 TI - Crystal structure of 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol kinase, an enzyme in the non-mevalonate pathway of isoprenoid synthesis. AB - The crystal structure of the enzyme 4-(cytidine 5'-diphospho)-2-C-methyl-D erythritol (CDP-ME) kinase from the thermophilic bacterium Thermus thermophilus HB8 has been determined at 1.7-A resolution. This enzyme catalyzes phosphorylation of the 2-hydroxyl group of CDP-ME, the fourth step of the non mevalonate pathway, which is essential for isoprenoid biosynthesis in several pathogenic microorganisms. Since this pathway is absent in humans, it is an important target for the development of novel antimicrobial compounds. The structure of the enzyme is similar to the structures of mevalonate kinase and homoserine kinase, members of the GHMP superfamily. Lys8 and Asp125 are active site residues in mevalonate kinase that also appear to play a catalytic role in CDP-ME kinase. Both the mevalonate and the non-mevalonate pathways therefore involve closely related kinases with similar mechanisms. Assaying the enzyme showed that CDP-ME kinase will phosphorylate CDP-ME but not 4-(uridine 5' diphospho)-2-C-methyl-D-erythritol, indicating the substrate pyrimidine moiety is involved in important interactions with the enzyme. PMID- 12771136 TI - Elucidation of the biosynthetic pathway of the allelochemical sorgoleone using retrobiosynthetic NMR analysis. AB - NMR analyses of the labeling pattern obtained using various 13C-labeled precursors indicated that both the lipid tail and the quinone head of sorgoleone, the main allelopathic component of the oily root exudate of Sorghum bicolor, were derived from acetate units, but that the two moieties were synthesized in different subcellular compartments. The 16:3 fatty acid precursor of the tail is synthesized by the combined action of fatty-acid synthase and desaturases most likely in the plastids. It is then exported out of the plastids and converted to 5-pentadecatriene resorcinol by a polyketide synthase. This resorcinol intermediate was identified in root hair extracts. The lipid resorcinol intermediate is then methylated by a S-adenosylmethionine-dependent O methyltransferase and subsequently dihydroxylated by a P450 monooxygenase to yield the reduced form of sorgoleone. PMID- 12771137 TI - Prolyl hydroxylation of collagen type I is required for efficient binding to integrin alpha 1 beta 1 and platelet glycoprotein VI but not to alpha 2 beta 1. AB - Collagen is a potent adhesive substrate for cells, an event essentially mediated by the integrins alpha 1 beta 1 and alpha 2 beta 1. Collagen fibrils also bind to the integrin alpha 2 beta 1 and the platelet receptor glycoprotein VI to activate and aggregate platelets. The distinct triple helical recognition motifs for these receptors, GXOGER and (GPO)n, respectively, all contain hydroxyproline. Using unhydroxylated collagen I produced in transgenic plants, we investigated the role of hydroxyproline in the receptor-binding properties of collagen. We show that alpha 2 beta 1 but not alpha 1 beta 1 mediates cell adhesion to unhydroxylated collagen. Soluble recombinant alpha 1 beta 1 binding to unhydroxylated collagen is considerably reduced compared with bovine collagens, but binding can be restored by prolyl hydroxylation of recombinant collagen. We also show that platelets use alpha 2 beta 1 to adhere to the unhydroxylated recombinant molecules, but the adhesion is weaker than on fully hydroxylated collagen, and the unhydroxylated collagen fibrils fail to aggregate platelets. Prolyl hydroxylation is thus required for binding of collagen to platelet glycoprotein VI and to cells by alpha 1 beta 1. These observations give new insights into the molecular basis of collagen-receptor interactions and offer new selective applications for the recombinant unhydroxylated collagen I. PMID- 12771138 TI - Unfolding of rabbit muscle creatine kinase induced by acid. A study using electrospray ionization mass spectrometry, isothermal titration calorimetry, and fluorescence spectroscopy. AB - Electrospray ionization mass spectrometry, isothermal titration calorimetry (ITC), fluorescence spectroscopy, and glutaraldehyde cross-linking SDS-PAGE have been used to study the unfolding of rabbit muscle creatine kinase (MM-CK) induced by acid. The mass spectrometric experiments show that MM-CK is unfolded gradually when titrated with acid. MM-CK is a dimer (the native state) at pH 7.0 and becomes an equilibrium mixture of the dimer and a partially folded monomer (the intermediate) between pH 6.7 and 5.0. The dimeric protein becomes an equilibrium mixture of the intermediate and an unfolded monomer (the unfolded state) between pH 5.0 and 3.0 and is almost fully unfolded at pH 3.0 reached. The results from a "phase diagram" method of fluorescence show that the conformational transition between the native state and the intermediate of MM-CK occurs in the pH range of 7.0-5.2, and the transition between the intermediate and the unfolded state of the protein occurs between pH 5.2 and 3.0. The intrinsic molar enthalpy changes for formation of the unfolded state of MM-CK induced by acid at 15.0, 25.0, 30.0, and 37.0 degrees C have been determined by ITC. A large positive molar heat capacity change of the unfolding, 8.78 kcal mol-1 K-1, at all temperatures examined indicates that hydrophobic interaction is the dominant driving force stabilizing the native structure of MM-CK. Combining the results from these four methods, we conclude that the acid-induced unfolding of MM-CK follows a "three state" model and that the intermediate state of the protein is a partially folded monomer. PMID- 12771139 TI - Hox proteins functionally cooperate with the GC box-binding protein system through distinct domains. AB - Hox genes encode a transcriptional factor that plays a key role in regulating position-specific cartilage pattern formation. We found that Hoxa-13 and Hoxd-13, which are members of the Abd-B subfamily of Hox genes and are crucial for the autopod development of the limb, stimulate transcription from the Bmp-4 promoter. This stimulation was dependent on the GC box within the promoter and independent of the putative Hox protein binding site. The stimulation by HoxA-13 was remarkably enhanced by cotransfection with members of a family of zinc finger GC box binding transcriptional factors including Sp1. The stimulation was suppressed by another Abd-B Hox protein, HoxA-11, indicating that each Abd-B Hox protein has a different effect on the target genes through the Sp1 system. We have identified multiple functional domains involved in transcriptional regulation, including three independent transcriptional activation domains (ADs) in HoxA-13. AD1 and AD3 in helices 1 and 2 of the homeodomain individually cooperate with Sp1 dependent stimulation. The homeodomain is also required for cooperation of the AD with Sp1. By contrast, AD2 strongly activates transcription in an Sp1-independent manner only when the homeodomain has been removed. These observations indicate that HoxA-13 regulates transcription through multiple pathways. In addition, we found that a helix 3 mutation of the HoxA-13 homeodomain behaves as a dominant negative form. PMID- 12771140 TI - Src kinase mediates the regulation of phospholipase C-gamma activity by glycosphingolipids. AB - Glucosylceramide-based glycosphingolipids have been previously demonstrated to regulate negatively the formation of inositol 1,4,5-trisphosphate by phospholipase C-gamma1. In the present study, the depletion of endogenous glucosylceramide by D-t-EtDO-P4 in cultured ECV304 cells induced autophosphorylation of Src kinase at tyrosine residue 418 within the catalytic loop and dephosphorylation of Src kinase at tyrosine residues 529 within the carboxyl-terminal regulatory region. Phosphotransferase activities of Src kinase were also induced in the glucosylceramide-depleted cells. c-Src kinase activity and phosphorylations at Src Tyr-418 and epidermal growth factor (EGF) receptor Tyr-1068 were significantly enhanced by bradykinin in response to 100 nm D-t-EtDO P4 compared with control cells. The phosphorylation and dephosphorylation on Tyr 418 and Tyr-529 residues of c-Src were reversed by treatment of 4-amino-5-(4 chlorophenyl)-7-t-butyl(pyrazolo)[3,4-d]pyrimidine (PP2), an inhibitor of Src kinase, in control cells. Glucosylceramide-depleted cells resisted treatment with PP2, and both phosphorylation of Tyr-418 and dephosphorylation of Tyr-529 induced by depletion of glucosylceramide were maintained. Compared with untreated cells, tyrosine phosphorylation of phospholipase C-gamma1 was enhanced by EGF stimulation in glucosylceramide-depleted cells, associated with enhanced tyrosine phosphorylation of the EGF receptor at Tyr-1068 and Tyr-1086 stimulated by EGF. The Src inhibitor, PP2, significantly blocked EGF-induced tyrosine phosphorylation of phospholipase C-gamma1 in control cells, whereas in glucosylceramide-depleted cells, suppression of Src kinase activity by PP2 toward EGF-induced tyrosine phosphorylation of phospholipase C-gamma1 was less significant. Thus the activation of Src kinase by depletion of glucosylceramide based glycosphingolipids in cultured ECV304 cells is a critical up-stream event in the activation of phospholipase C-gamma1. PMID- 12771141 TI - Structural basis of Synercid (quinupristin-dalfopristin) resistance in Gram positive bacterial pathogens. AB - Synercid, a new semisynthetic streptogramin-derived antibiotic containing dalfopristin and quinupristin, is used in treatment of life-threatening infections caused by glycopeptide-resistant Enterococcus faecium and other bacterial pathogens. However, dissemination of genes encoding virginiamycin acetyltransferases, enzymes that confer resistance to streptogramins, threatens to limit the medical utility of the quinupristin-dalfopristin combination. Here we present structures of virginiamycin acetyltransferase D (VatD) determined at 1.8 A resolution in the absence of ligands, at 2.8 A resolution bound to dalfopristin, and at 3.0 A resolution in the presence of acetyl-coenzyme A. Dalfopristin is bound by VatD in a similar conformation to that described previously for the streptogramin virginiamycin M1. However, specific interactions with the substrate are altered as a consequence of a conformational change in the pyrollidine ring that is propagated to adjacent constituents of the dalfopristin macrocycle. Inactivation of dalfopristin involves acetyl transfer from acetyl coenzyme A to the sole (O-18) hydroxy group of the antibiotic that lies close to the side chain of the strictly conserved residue, His-82. Replacement of residue 82 by alanine is accompanied by a fall in specific activity of >105-fold, indicating that the imidazole moiety of His-82 is a major determinant of catalytic rate enhancement by VatD. The structure of the VatD-dalfopristin complex can be used to predict positions where further structural modification of the drug might preclude enzyme binding and thereby circumvent Synercid resistance. PMID- 12771142 TI - Src kinases mediate STAT growth pathways in squamous cell carcinoma of the head and neck. AB - Signal transducer and activator of transcription (STAT) proteins are constitutively activated in many malignancies, including squamous cell carcinoma of the head and neck (SCCHN). Previously, we reported that phosphorylation of the epidermal growth factor receptor (EGFR) is linked to activation of STATs 3 and 5 in SCCHN cells. The present study was undertaken to determine the role of Src family kinases in STAT activation and SCCHN growth. The Src family kinases c-Src, c-Yes, Fyn, and Lyn were expressed and activated by transforming growth factor alpha stimulation in all four SCCHN cell lines examined but not in corresponding normal epithelial cells. In nine SCCHN cell lines tested, Src phosphotyrosine expression levels were highly correlated with activation levels of STATs 3 and 5. Co-immunoprecipitation analysis demonstrated interaction between c-Src and STATs 3 or 5 and EGFR in SCCHN cells, but no heterodimerization was detected between STAT3 and STAT5. SCCHN cells treated with either of two Src-specific inhibitors or transfected with a dominant-negative c-Src construct demonstrated decreased activation of STATs 3 and 5 and reduced growth rates in vitro. These results demonstrate a role for Src kinases in mediating activation of STATs 3 and 5 in concert with the EGFR in SCCHN cells. Strategies to target Src activation may contribute to the treatment of cancers that demonstrate increased levels of EGFR and STATs, including SCCHN. PMID- 12771143 TI - Direct demonstration of ATP-dependent release of SecA from a translocating preprotein by surface plasmon resonance. AB - Translocase mediates the transport of preproteins across the inner membrane of Escherichia coli. SecA binds with high affinity to the membrane-embedded protein conducting SecYEG complex and serves as both a receptor for secretory proteins and as an ATP-driven molecular motor. Cycles of ATP binding and hydrolysis by SecA drive the progressive movement of the preprotein across the membrane. Surface plasmon resonance allows an online monitoring of protein interactions. Here we report on the kinetic analysis of the interaction between SecA and the membrane-embedded SecYEG complex. Immobilization of membrane vesicles containing overproduced SecYEG on the Biacore Pioneer L1 chip allows the detection of high affinity SecA binding to the SecYEG complex and online monitoring of the translocation of the secretory protein proOmpA. SecA binds tightly to the SecYEG.proOmpA complex and is released only upon ATP hydrolysis. The results provide direct evidence for a model in which SecA cycles at the SecYEG complex during translocation. PMID- 12771144 TI - Osteopontin stimulates cell motility and nuclear factor kappaB-mediated secretion of urokinase type plasminogen activator through phosphatidylinositol 3-kinase/Akt signaling pathways in breast cancer cells. AB - We have recently reported that osteopontin (OPN) induces nuclear factor kappaB (NFkappaB)-mediated promatrix metalloproteinase-2 activation through IkappaBalpha/IKK signaling pathways and that curcumin (diferulolylmethane) down regulates these pathways (Philip, S., and Kundu, G. C. (2003) J. Biol. Chem. 278, 14487-14497). However, the molecular mechanism by which upstream kinases regulate the OPN-induced NFkappaB activation and urokinase type plasminogen activator (uPA) secretion in human breast cancer cells is not well defined. Here we report that OPN induces the phosphatidylinositol 3'-kinase (PI 3'-kinase) activity and phosphorylation of Akt in highly invasive MDA-MB-231 and low invasive MCF-7 cells. The OPN-induced Akt phosphorylation was inhibited when cells were transfected with a dominant negative mutant of the p85 domain of PI 3-kinase (Deltap85) and enhanced when cells were transfected with an activated form of PI 3-kinase (p110CAAX), indicating that PI 3'-kinase is involved in Akt phosphorylation. OPN enhances the interaction between IkappaBalpha kinase (IKK) and phosphorylated Akt. OPN also induces NFkappaB activation through phosphorylation and degradation of IkappaBalpha by inducing the IKK activity. However, both pharmacological (wortmannin and LY294002) and genetic (Deltap85) inhibitors of PI 3'-kinase inhibited OPN-induced Akt phosphorylation, IKK activity, and NFkappaB activation through phosphorylation and degradation of IkappaBalpha. OPN also enhances uPA secretion, cell motility, and extracellular matrix invasion. Furthermore, cells transfected with Deltap85 or the super repressor form of IkappaBalpha suppressed the OPN-induced uPA secretion and cell motility, whereas cells transfected with p110CAAX enhanced these effects. Pretreatment of cells with PI 3-kinase inhibitors or NFkappaB inhibitory peptide (SN-50) reduced the OPN-induced uPA secretion, cell motility, and invasion. To our knowledge, this is first report that OPN induces NFkappaB activity and uPA secretion by activating PI 3'-kinase/Akt/IKK-mediated signaling pathways and further demonstrates a functional molecular link between OPN-induced PI 3'-kinase dependent Akt phosphorylation and NFkappaB-mediated uPA secretion, and all of these ultimately control the motility of breast cancer cells. PMID- 12771145 TI - Palmitate inhibition of insulin gene expression is mediated at the transcriptional level via ceramide synthesis. AB - Chronic exposure to elevated levels of fatty acids impairs pancreatic beta cell function, a phenomenon thought to contribute to the progressive deterioration of insulin secretion in type 2 diabetes. We have previously demonstrated that prolonged exposure of isolated islets to elevated levels of palmitate inhibits preproinsulin mRNA levels in the presence of high glucose concentrations. However, whether this occurs via transcriptional or post-transcriptional mechanisms has not been determined. In addition, the nature of the lipid metabolites involved in palmitate inhibition of insulin gene expression is unknown. In this study, we show that palmitate decreases glucose-stimulated preproinsulin mRNA levels in isolated rat islets, an effect that is not mediated by changes in preproinsulin mRNA stability, but is associated with inhibition of glucose-stimulated insulin promoter activity. Prolonged culture of isolated islets with palmitate is associated with increased levels of intracellular ceramide. Palmitate-induced ceramide generation is prevented by inhibitors of de novo ceramide synthesis. Further, exogenous ceramide inhibits insulin mRNA levels, whereas blockade of de novo ceramide synthesis prevents palmitate inhibition of insulin gene expression. We conclude that prolonged exposure to elevated levels of palmitate affects glucose-stimulated insulin gene expression via transcriptional mechanisms and ceramide synthesis. PMID- 12771146 TI - The tyrosine kinase Pyk2 regulates Arf1 activity by phosphorylation and inhibition of the Arf-GTPase-activating protein ASAP1. AB - Proline-rich tyrosine kinase 2 (Pyk2), a non-receptor tyrosine kinase structurally related to focal adhesion kinase, has been implicated in the regulation of mitogen-activated protein kinase cascades and ion channels, the induction of apoptosis, and in the modulation of the cytoskeleton. In order to understand how Pyk2 signaling mediates these diverse cellular functions, we performed a yeast two-hybrid screening using the C-terminal part of Pyk2 that contains potential protein-protein interaction sites as bait. A prominent binder of Pyk2 identified by this method was the Arf-GTPase-activating protein ASAP1. Pyk2-ASAP1 interaction was confirmed in pull-down as well as in co immunoprecipitation experiments, and contact sites were mapped to the proline rich regions of Pyk2 and the SH3 domain of ASAP1. Pyk2 directly phosphorylates ASAP1 on tyrosine residues in vitro and increases ASAP1 tyrosine phosphorylation when co-expressed in HEK293T cells. Phosphorylation of tyrosine 308 and 782 affects the phosphoinositide binding profile of ASAP1, and fluorimetric Arf GTPase assays with purified proteins revealed an inhibition of ASAP1 GTPase activating protein activity by Pyk2-mediated tyrosine phosphorylation. We therefore provide evidence for a functional interaction between Pyk2 and ASAP1 and a regulation of ASAP1 and hence Arf1 activity by Pyk2-mediated tyrosine phosphorylation. PMID- 12771147 TI - Eukaryotic NAD+ synthetase Qns1 contains an essential, obligate intramolecular thiol glutamine amidotransferase domain related to nitrilase. AB - NAD+ is an essential co-enzyme for redox reactions and is consumed in lysine deacetylation and poly(ADP-ribosyl)ation. NAD+ synthetase catalyzes the final step in NAD+ synthesis in the well characterized de novo, salvage, and import pathways. It has been long known that eukaryotic NAD+ synthetases use glutamine to amidate nicotinic acid adenine dinucleotide while many purified prokaryotic NAD+ synthetases are ammonia-dependent. Earlier, we discovered that glutamine dependent NAD+ synthetases contain N-terminal domains that are members of the nitrilase superfamily and hypothesized that these domains function as glutamine amidotransferases for the associated synthetases. Here we show yeast glutamine dependent NAD+ synthetase Qns1 requires both the nitrilase-related active-site residues and the NAD+ synthetase active-site residues for function in vivo. Despite failure to complement the lethal phenotype of qns1 disruption, the former mutants retain ammonia-dependent NAD+ synthetase activity in vitro, whereas the latter mutants retain basal glutaminase activity. Moreover, the two classes of mutants fail to trans-complement despite forming a stable heteromultimer in vivo. These data indicate that the nitrilase-related domain in Qns1 is the fourth independently evolved glutamine amidotransferase domain to have been identified in nature and that glutamine-dependence is an obligate phenomenon involving intramolecular transfer of ammonia over a predicted distance of 46 A from one active site to another within Qns1 monomers. PMID- 12771148 TI - Signal sequences initiate the pathway of maturation in the endoplasmic reticulum lumen. AB - An interaction between an N-terminal signal sequence and the translocon leads to the initiation of protein translocation into the endoplasmic reticulum lumen. Subsequently, folding and modification of the substrate rapidly ensue. The close temporal coordination of these processes suggests that they may be structurally and functionally coordinated as well. Here we show that information encoded in the hydrophobic domain of a signal sequence influences the timing and efficiency of at least two steps in maturation, namely N-linked glycosylation and signal sequence cleavage. We demonstrate that these consequences correlate with and likely stem from the nature of the initial association made between the signal sequence and the translocon during the initiation of translocation. We propose a model by which these maturational events are controlled by the signal sequence translocon interaction. Our work demonstrates that the pathway taken by a nascent chain through post-translational maturation depends on information encoded in its signal sequence. PMID- 12771149 TI - Src kinase regulates the activation of a novel FGD-1-related Cdc42 guanine nucleotide exchange factor in the signaling pathway from the endothelin A receptor to JNK. AB - Small GTPases act as binary switches by cycling between an inactive (GDP-bound) and an active (GTP-bound) state. Upon stimulation with extracellular signals, guanine-nucleotide exchange factors (GEFs) stimulate the exchange of GDP to GTP to shift toward the active forms of small GTPases, recognizing the downstream targets. Here we show that KIAA0793, containing substantial sequence homology with the catalytic Dbl homology domain of the faciogenital dysplasia gene product (FGD1), is a specific GEF for Cdc42. We, therefore, tentatively named it FRG (FGD1-related Cdc42-GEF). Src kinase directly phosphorylates and activates FRG, as Vav family GEFs. Additionally, FRG is involved in the signaling pathway from the endothelin A receptor to c-Jun N-terminal kinase, resulting in the inhibition of cell motility. These results suggest that FRG is a member of Cdc42-GEF and plays an important role in the signaling pathway downstream of G protein-coupled receptors. PMID- 12771150 TI - A novel cyclic peptide immunization strategy for preventing HIV-1/AIDS infection and progression. AB - A novel synthetic peptide immunogen targeting the human immunodeficiency virus type-1 (HIV-1) coreceptor CXCR4 was evaluated for its capacity to induce CXCR4 specific antibodies with anti-HIV-1 activity in BALB/c mice and cynomolgus monkeys. A cyclic closed-chain dodecapeptide mimicking the conformation-specific domain of CXCR4 (cDDX4) was prepared in which Gly-Asp, as the dipeptide forming a spacer arm, links the amino and carboxyl termini of the decapeptidyl linear chain (linear DDX4, Asn176 to Ile185) derived from the undecapeptidyl arch (UPA; Asn176 to Cys186) of extracellular loop 2 (ECL-2) in CXCR4. Immunization of BALB/c mice with cDDX4 conjugated with a multiple-antigen peptide (cDDX4-MAP) induced conformational epitope-specific antibodies, and monoclonal antibody IA2-F9 reacted with cDDX4, but not with linear DDX4, as determined by real-time biomolecular interaction analysis using surface plasmon resonance. The antibody also reacted with cells expressing CXCR4 but not with cells expressing the other HIV coreceptor, CCR5. Furthermore, the antibody inhibited the replication of HIV 1 X4 virus (using CXCR4), as shown by an infection assay using both MAGIC-5 cells and MT4 cells, but not that of HIV-1 R5 virus (using CCR5). The antibody weakly interfered with chemotaxis induced by stromal cell-derived factor-1 alpha in THP 1 cells or moderately inhibited the chemotaxis of Molt4#8 cells under the same conditions. In addition, immunization of cynomolgus monkeys also induced cDDX4 specific antibodies with anti-HIV activity. Taken together, these results indicate that cDDX4 conjugated with a multi-antigen peptide induces the conformational epitope-specific antibodies to the undecapeptidyl arch of CXCR4 may be a novel candidate immunogen for preventing disease progression in HIV-1 infected individuals. PMID- 12771151 TI - Alanine-scanning mutations in domain 4 of anthrax toxin protective antigen reveal residues important for binding to the cellular receptor and to a neutralizing monoclonal antibody. AB - A panel of variants with alanine substitutions in the small loop of anthrax toxin protective antigen domain 4 was created to determine individual amino acid residues critical for interactions with the cellular receptor and with a neutralizing monoclonal antibody, 14B7. Substituted protective antigen proteins were analyzed by cellular cytotoxicity assays, and their interactions with antibody were measured by plasmon surface resonance and analytical ultracentrifugation. Residue Asp683 was the most critical for cell binding and toxicity, causing an approximately 1000-fold reduction in toxicity, but was not a large factor for interactions with 14B7. Substitutions in residues Tyr681, Asn682, and Pro686 also reduced toxicity significantly, by 10-100-fold. Of these, only Asn682 and Pro686 were also critical for interactions with 14B7. However, residues Lys684, Leu685, Leu687, and Tyr688 were critical for 14B7 binding without greatly affecting toxicity. The K684A and L685A variants exhibited wild type levels of toxicity in cell culture assays; the L687A and Y688A variants were reduced only 1.5- and 5-fold, respectively. PMID- 12771152 TI - Transforming growth factor-alpha attenuates N-methyl-D-aspartic acid toxicity in cortical cultures by preventing protein synthesis inhibition through an Erk1/2 dependent mechanism. AB - Transforming growth factor-alpha (TGF-alpha), a ligand of the epidermal growth factor receptor, reduces the infarct size after focal cerebral ischemia in rat, but the molecular basis underlying the protection is unknown. Excitotoxicity and global inhibition of translation are acknowledged to contribute significantly to the ischemic damage. Here we studied whether TGF-alpha can rescue neurons from excitotoxicity in vitro and how it affects calcium homeostasis, protein synthesis, and the associated Akt and extracellular signal-regulated kinase 1/2 (Erk1/2) intracellular signaling pathways in mixed neuron-glia cortical cultures. We found that 100 ng/ml TGF-alpha attenuated neuronal cell death induced by a 30 min exposure to 35 microM N-methyl-D-aspartic acid (NMDA) (as it reduced lactate dehydrogenase release, propidium iodide staining, and caspase-3 activation) and decreased the elevation of intracellular Ca2+ elicited by NMDA. TGF-alpha induced a prompt and sustained phosphorylation of Erk1/2 and prevented the loss of Akt-P induced by NMDA 3 h after exposure. The protective effect of TGF-alpha was completely prevented by PD 98059, an inhibitor of the Erk1/2 pathway. Studies of incorporation of [3H]leucine into proteins showed that NMDA decreased the rate of protein synthesis, and TGF-alpha attenuated this effect. TGF-alpha stimulated the phosphorylation of the eukaryotic initiation factor 4E (eIF4E) but did not affect eIF2 alpha, two proteins involved in translation regulation. PD 98059 abrogated the TGF-alpha effect on eIF4E. Our data demonstrate that TGF-alpha exerts a neuroprotective action against NMDA toxicity, in which Erk1/2 activation plays a key role, and suggest that the underlying mechanisms involve recovery of translation inhibition, mediated at least in part by eIF4E phosphorylation. PMID- 12771153 TI - Two novel proteins that are linked to insulin-like growth factor (IGF-I) receptors by the Grb10 adapter and modulate IGF-I signaling. AB - Grb10 is a protein that binds to the intracellular domains of activated tyrosine kinase receptors, including insulin-like growth factor (IGF-I) and insulin receptors. This occurs through the interaction of two C-terminal Grb10 motifs (BPS and Src homology domains) with receptor phosphotyrosine residues. Published data from transfection/overexpression studies support both positive and negative regulatory effects of Grb10, thus leaving its physiological role unclear. Because Grb10 has the structure of an adapter protein, the objective of this study was to determine whether Grb10 links other proteins to IGF-I receptors and thus modulates IGF-I signaling. Using yeast two-hybrid screening, the N terminus of Grb10 was shown to interact with two novel proteins, designated GIGYF1 (Grb10 interacting GYF protein 1) and GIGYF2. Mutation analysis indicates that a 17 amino acid sequence in GIGYF1 and GIGYF2, homologous to the GYF domain described previously, binds to tandem proline-rich regions in the N terminus of Grb10. In IGF-I receptor-expressing R+ fibroblasts, there is detectable binding of a Myc tagged fragment of GIGYF1 to Grb10 in the basal state. Stimulation with IGF-I results in increased binding of GIGYF1 to Grb10 and transient binding of both Grb10 and GIGYF1 to IGF-I receptors, presumably via the adapter function of Grb10. At later time points, GIGYF1 dissociates, but Grb10 remains linked to IGF I receptors. Overexpression of the Grb10 binding fragment of GIGYF1 in R+ cells results in a significant increase in IGF-I-stimulated receptor tyrosine phosphorylation. In conclusion, we have identified two members of a novel protein family, which become transiently linked to IGF-I receptors by the Grb10 adapter protein following IGF-I stimulation. Grb10 and GIGYFs may act cooperatively to regulate receptor signaling. PMID- 12771154 TI - The prodomain of a secreted hydrophobic mini-protein facilitates its export from the endoplasmic reticulum by hitchhiking on sorting receptors. AB - Misfolded secretory proteins are retained in the endoplasmic reticulum (ER) by quality control mechanisms targeted to exposed hydrophobic surfaces. Paradoxically, certain conotoxins expose extensive hydrophobic surfaces upon folding to their bioactive structures. How then can such secreted mini-proteins traverse the secretory pathway? Here we show that secretion of the hydrophobic conotoxin-TxVI is strongly dependent on its propeptide domain, which enhances TxVI export from the ER. The propeptide domain interacts with sorting receptors from the sortilin Vps10p domain family. The sortilin-TxVI interaction occurs in the ER, and sortilin facilitates export of TxVI from the ER to the Golgi. Thus, the prodomain in a secreted hydrophobic protein acts as a tag that can facilitate its ER export by a hitchhiking mechanism. PMID- 12771155 TI - Receptor-dependent RhoA activation in G12/G13-deficient cells: genetic evidence for an involvement of Gq/G11. AB - The small GTPase RhoA is involved in the regulation of various cellular functions like the remodeling of the actin cytoskeleton and the induction of transcriptional activity. G-protein-coupled receptors (GPCRs), which are able to activate Gq/G11 and G12/G13 are major upstream regulators of RhoA activity, and G12/G13 have been shown to couple GPCRs to the activation of Rho by regulating the activity of a subfamily of RhoGEF proteins. However, the possible contribution of Gq/G11 to the regulation of RhoA activity via GPCRs is controversial. We have used a genetic approach to study the role of heterotrimeric G-proteins in the activation of RhoA via endogenous GPCRs. In pertussis toxin-treated Galpha12/Galpha13-deficient as well as in Galphaq/Galpha11-deficient mouse embryonic fibroblasts (MEFs), in which coupling of receptors is restricted to Gq/G11 and G12/G13, respectively, receptor activation results in Rho activation. Rho activation induced by receptor agonists via Gq/G11 occurs with lower potency than Rho activation via G12/G13. Activation of RhoA via Gq/G11 is not affected by the phospholipase-C blocker U73122 or the Ca2+-chelator BAPTA, but can be blocked by a dominant-negative mutant of the RhoGEF protein LARG. Our data clearly show that G12/G13 as well as Gq/G11 alone can couple GPCRs to the rapid activation of RhoA. Gq/G11-mediated RhoA activation occurs independently of phospholipase C-beta and appears to involve LARG. PMID- 12771156 TI - Dimethyl sulfoxide exposure facilitates phospholipid biosynthesis and cellular membrane proliferation in yeast cells. AB - Me2SO is a polar solvent that is widely used in biochemistry, pharmacology, and industry. Although there are several reports in the literature concerning the biological effects of Me2SO, the total cellular response remains unclear. In this paper, DNA microarray technology combined with the hierarchical clustering bioinformatics tool was used to assess the effects of Me2SO on yeast cells. We found that yeast exposed to Me2SO increased phospholipid biosynthesis through up regulated gene expression. It was confirmed by Northern blotting that the level of INO1 and OPI3 gene transcripts, encoding key enzymes in phospholipid biosynthesis, were significantly elevated following treatment with Me2SO. Furthermore, the phospholipid content of the cells increased during exposure to Me2SO as shown by conspicuous incorporation of a lipophilic fluorescent dye (3,3' dihexyloxacarbocyanine iodide) into the cell membranes. From these results we propose that Me2SO treatment induces membrane proliferation in yeast cells to alleviate the adverse affects of this chemical on membrane integrity. PMID- 12771158 TI - Associative learning and memory in Lymnaea stagnalis: how well do they remember? AB - The search for 'the how and the where' of memory formation in the brain, the engram, is still one of the unattained 'Holy Grails' of neuroscience. Over the years, various paths have been trodden in attempts to attain this goal, and while tantalizing glimpses appear now and then on the scientific horizon, the Grail still has not been grasped. One of the paths that investigators have walked is the invertebrate 'model system' approach. Some invertebrates possess relatively simple nervous systems that mediate relatively simple behaviours that are both interesting and trainable. In this commentary, we would like to shed light on a relatively new player, the pond snail Lymnaea stagnalis L., that is being used in the quest to illuminate 'the how and the where' the nervous systems encode and store memory. We will show that it is possible to demonstrate that a single neuron is a site of memory formation and storage for a form of associative learning in this lowly snail. It may be that the Grail is a little closer to being grasped. PMID- 12771159 TI - Visual resolution of gratings by the compound eye of the bee Apis mellifera. AB - Early measurements of the resolution of horizontal versus vertical gratings were confirmed, with a limit near a period of 2.5 degrees, and the resolution is similar when vertical or horizontal gratings are tested separately against grey. Bees were next trained to discriminate from a distance between gratings at 45 degrees versus 135 degrees, with no green contrast, on targets presented in a vertical plane at a fixed distance. As expected, they fail to learn; however, with green contrast but no modulation difference the resolution limit is near 3.5 degrees. With vertical and horizontal gratings with no green contrast they discriminate but do not learn an orientation cue. In order to eliminate the orientation cue altogether, new bees were then trained with alternating vertical and horizontal gratings versus grey, or with a black and white checkerboard versus grey. Tests of these trained bees with horizontal or with vertical gratings separately against grey again show a resolution down to a period near 2.5 degrees. These results, taken together, show that when edge orientation alone is the cue, the limit of resolution is near 3.5 degrees, but when receptor modulation is the cue, the limit is near 2.5 degrees. PMID- 12771160 TI - The tremble dance of honey bees can be caused by hive-external foraging experience. AB - The tremble dance of honey bee nectar foragers is part of the communication system that regulates a colony's foraging efficiency. A forager that returns to the hive with nectar, but then experiences a long unloading delay because she has difficulty finding a nectar receiver bee, will perform a tremble dance to recruit additional nectar receiver bees. A forager that experiences a short unloading delay will perform a waggle dance to recruit more nectar foragers. A long unloading delay was until now the only known cause of tremble dancing. However, several studies suggested that factors at the food source may also cause tremble dancing. Here I test whether one of these factors, crowding of nectar foragers at the food source, stimulates tremble dancing because it causes long unloading delays. To do so, I increased the density of nectar foragers at a food source by suddenly reducing the size of an artificial feeder, and recorded the unloading delay experienced by each forager, as well as the dance she performed, if any. A forager's unloading delay was measured as the time interval between entering the hive and either (1) the first unloading contact with a nectar receiver bee, or (2) the start of the first dance, if dancing began before the first unloading contact. I also recorded the unloading delays and dances of nectar foragers that returned from natural food sources. The results show that crowding of nectar foragers at the food source increases the probability of tremble dancing, but does not cause long unloading delays, and that tremble dancers that foraged at natural food sources also often have short unloading delays. When the cause of the tremble dance is not a low supply of nectar receiver bees, the tremble dance may have a function in addition to the recruitment of nectar receiver bees. PMID- 12771161 TI - The effects of gape angle and bite point on bite force in bats. AB - Models of mammalian mastication predict that bite force is affected by both the degree of mouth opening (gape angle) and the point along the tooth row at which force is transferred to a food item (bite point). Despite the widespread use of these models in comparative analyses, experimental data documenting bite force in non-human mammals are extremely limited. The goal of this study is to document variation in non-stimulated bite force associated with change in gape angle and bite point in a broad range of species. We focus on plant-visiting bats because they exhibit a relatively primitive cranial morphology and are good models for generalized mammals. Assessments of the relationship between gape angle and bite force within and among species demonstrate that bite force decreases significantly as gape angle increases. The relationship between bite force and bite point within each of seven species demonstrates that unilateral molar biting universally generates the highest forces while the unilateral canine biting produces the lowest forces. Bilateral canine biting is intermediate. Beyond these general patterns, differences among species suggest that bite force reflects variation in craniofacial architecture. Finally, these data suggest that behavioral variation in gape angle and bite point may be important variables in comparative, functional analyses of feeding. PMID- 12771162 TI - Energetics of nestling growth and parental effort in Antarctic fulmarine petrels. AB - Antarctic fulmarine petrels breed in some of the coldest conditions encountered by any bird and their young grow twice as fast as predicted allometrically. To examine the energetic consequences of fast growth in a cold environment, we used the doubly labeled water technique to measure field metabolic rates of adults (three species) and different-aged nestlings (four species) of Antarctic fulmarine petrels in the Rauer Islands, East Antarctica: Antarctic fulmar Fulmarus glacialoides, Antarctic petrel Thalassoica antarctica, Cape petrel Daption capense and snow petrel Pagodroma nivea. We used our data to assess parental effort and, together with literature values on nestling growth and resting metabolic rate, to construct and partition nestling energy budgets. Nestling total energy expenditure and peak daily metabolic rate were significantly higher than predicted allometrically (33-73% and 17-66% higher, respectively), and the relative cost of growth in nestling petrels was among the highest reported for birds (54-72 kJ g(-1)). Parental effort during the nestling feeding period was identical in adult Cape and Antarctic petrels (3.5 times basal metabolic rate, BMR), and was somewhat (but not significantly) higher in snow petrels (4.6 times BMR). These values are comparable to those of other high latitude procellariiform birds. Thus, despite the constraints of a compressed breeding season, cold temperatures and fast-growing nestlings, adult Antarctic fulmarine petrels do not work harder than procellariid adults whose chicks grow much more slowly. Our findings suggest that obtaining sufficient food is generally not a constraint for adult fulmarine petrels and that factors operating at the tissue level limit nestling growth rate. PMID- 12771163 TI - Role of the post-hepatic septum on breathing during locomotion in Tupinambis merianae (Reptilia: Teiidae). AB - Tupinambis merianae increased minute ventilation by increasing both tidal volume and breathing frequency during sustained locomotion at 0.17 m s(-1). Animals in which the post-hepatic septum (PHS) had been surgically removed were not able to increase tidal volume during locomotion. Tegus without PHS compensated, in part, by increasing breathing frequency above the levels observed for tegus with intact PHS, but minute ventilation remained less than in the control animals. The rate of oxygen consumption and the air convection requirement, however, were not significantly different between animals with and without PHS, nor at the tested speeds was endurance affected by the removal of the PHS. These data suggest that the PHS facilitates ventilation by acting as a mechanical barrier, preventing the viscera from moving cranially during physical exertion. PMID- 12771164 TI - Body size as a latent variable in a structural equation model: thermal acclimation and energetics of the leaf-eared mouse. AB - Body size is one of the most important determinants of energy metabolism in mammals. However, the usual physiological variables measured to characterize energy metabolism and heat dissipation in endotherms are strongly affected by thermal acclimation, and are also correlated among themselves. In addition to choosing the appropriate measurement of body size, these problems create additional complications when analyzing the relationships among physiological variables such as basal metabolism, non-shivering thermogenesis, thermoregulatory maximum metabolic rate and minimum thermal conductance, body size dependence, and the effect of thermal acclimation on them. We measured these variables in Phyllotis darwini, a murid rodent from central Chile, under conditions of warm and cold acclimation. In addition to standard statistical analyses to determine the effect of thermal acclimation on each variable and the body-mass-controlled correlation among them, we performed a Structural Equation Modeling analysis to evaluate the effects of three different measurements of body size (body mass, m(b); body length, L(b) and foot length, L(f)) on energy metabolism and thermal conductance. We found that thermal acclimation changed the correlation among physiological variables. Only cold-acclimated animals supported our a priori path models, and m(b) appeared to be the best descriptor of body size (compared with L(b) and L(f)) when dealing with energy metabolism and thermal conductance. However, while m(b) appeared to be the strongest determinant of energy metabolism, there was an important and significant contribution of L(b) (but not L(f)) to thermal conductance. This study demonstrates how additional information can be drawn from physiological ecology and general organismal studies by applying Structural Equation Modeling when multiple variables are measured in the same individuals. PMID- 12771165 TI - Repeated bouts of high-intensity exercise and muscle glycogen sparing in the rat. AB - Even in the absence of food intake, several animal species recovering from physical activity of high intensity can replenish completely their muscle glycogen stores. In some species of mammals, such as in rats and humans, glycogen repletion is only partial, thus suggesting that a few consecutive bouts of high intensity exercise might eventually lead to the sustained depletion of their muscle glycogen. In order to test this prediction, groups of rats with a lead weight of 10% body mass attached to their tails were subjected to either one, two or three bouts of high-intensity swimming, each bout being separated from the next by a 1 h re covery period. Although glycogen repletion after the first bout of exercise was only partial, all the glycogen mobilised in subsequent bouts was completely replenished during the corresponding recovery periods and irrespective of muscle fibre compositions. The impact of repeated bouts of high-intensity exercise on plasma levels of fatty acids, acetoacetate and beta-hydroxybutyrate suggests that the metabolic state of the rat prior to the second and third bouts of exercise was different from that before the first bout. In conclusion, rats resemble other vertebrate species in that without food intake there are conditions under which they can replenish completely their muscle glycogen stores from endogenous carbon sources when recovering from high-intensity exercise. It remains to be established, however, whether this capacity is typical of mammals in general. PMID- 12771166 TI - Molecular characterisation of the smooth endoplasmic reticulum Ca(2+)-ATPase of Porcellio scaber and its expression in sternal epithelia during the moult cycle. AB - The anterior sternal epithelial cells of the terrestrial isopod Porcellio scaber transport large amounts of calcium during the formation and resorption of intermittent calcium carbonate deposits. Recent investigations on epithelia involved in mineralisation processes suggest a role of the smooth endoplasmic reticulum Ca(2+)-ATPase (SERCA) in transcellular calcium transport. We present the first molecular characterisation of a SERCA within a crustacean mineralising epithelium. We cloned the SERCA from a cDNA library of the anterior sternal epithelium and used in situ hybridisation to compare the expression of the SERCA mRNA between three different moulting stages. The full-length SERCA cDNA has an open reading frame of 3006 nucleotides. The deduced 1002 amino-acid polypeptide has a predicted molecular mass of 109.7 kDa and 87% identity to the SERCA of Procambarus clarkii axial muscle isoform. In situ hybridisation confirmed expression within the anterior sternal epithelium and revealed an increase in SERCA mRNA abundance from the non-transporting, early premoult stage to the calcium transporting, late premoult and intramoult stage. The results support previous indications of a contribution by the smooth endoplasmic reticulum to transcellular calcium transport and suggest a transcriptional regulation of SERCA activity. PMID- 12771167 TI - Occlusable corneas in toadfishes: light transmission, movement and ultrastruture of pigment during light- and dark-adaptation. AB - The toadfishes Tetractenos hamiltoni and Torquigener pleurogramma (Tetraodontidae) possess occlusable yellow corneas. We examine the light transmission and location of the yellow/orange pigment throughout the cornea, the temporal properties of pigment migration and the ultrastructure of the pigmented processes during light- and dark-adaptation. Each species was dark-adapted during the day and light-adapted during the night and then exposed to either sun illumination or darkness for different lengths of time (0-70 min). Movement of corneal pigment could be induced in both species regardless of time of day or night. The pigment was able to migrate in a dorsal or ventral direction and changed from minimal to maximal pigmentation within 60 min. Three types of transmission curves were found with varying degrees of transmission in the 400 500 nm waveband, indicating that the pigment distribution is not uniform across the cornea; some areas of the cornea transmit near UV light, while others absorb blue light. The gradual change of the transmission characteristics in different areas of the cornea indicates the presence of different concentrations of a single type of pigment. Ultrastructural examination of the corneas showed that the layer containing the pigment is situated within the scleral cornea either surrounding (T. pleurogramma) or abutting (T. hamiltoni) an iridescent layer. Long sheet-like processes or chromatophores extending centrally from dorsal and ventral reservoirs are filled with pigment during the light-adapted state but empty in the dark-adapted state. PMID- 12771168 TI - Neurotrophin 4/5 is required for the normal development of the slow muscle fiber phenotype in the rat soleus. AB - During normal postnatal development, rat soleus (SOL) muscle fibers undergo a dramatic fast-to-slow myosin heavy chain (MyHC) isoform transformation. We exploited this phenomenon to evaluate the role of neurotrophin 4/5 (NT-4/5) in the regulation of muscle fiber phenotype. Intramuscular injections of recombinant NT-4/5 into the SOL muscle of rat neonates significantly accelerated the normal fast-to-slow MyHC isoform transformation. Sequestration of endogenous NT-4/5 with TrkB-IgG prevented this transformation from occurring. Administration of the other TrkB ligand, brain-derived neurotrophic factor (BDNF), did not affect the normal course of the MyHC isoform transformation in this muscle, indicating that the observed effect is NT-4/5 specific. Botulinum toxin blockade of synaptic transmission significantly disrupted the normal fast-to-slow MyHC isoform switch. Because administration of NT-4/5 to paralyzed muscles failed to restore the normal course of this MyHC transformation, we believe that the effect of NT-4/5 is not directly on the muscle fibers but that it probably activates or forms a type of retrograde signal to motoneurons. The developmental upregulation of NT 4/5 mRNA in rat SOL muscle fibers occurred earlier than the upregulation of MyHC I/b mRNA associated with muscle fiber transformation. This timing is consistent with the idea that NT-4/5 is involved in early events that lead to the upregulation of the slow MyHC isoform in this muscle. PMID- 12771169 TI - Olfactory input increases visual sensitivity in zebrafish: a possible function for the terminal nerve and dopaminergic interplexiform cells. AB - Centrifugal innervation of the neural retina has been documented in many species. In zebrafish Danio rerio, the only so-far described centrifugal pathway originates from terminal nerve (TN) cell bodies that are located in the olfactory bulb. Most of the TN axons terminate in the forebrain and midbrain, but some project via the optic nerve to the neural retina, where they synapse onto dopaminergic interplexiform cells (DA-IPCs). While the anatomical pathway between the olfactory and visual organs has been described, it is unknown if and how olfactory signals influence visual system functions. We demonstrate here that olfactory input is involved in the modulation of visual sensitivity in zebrafish. As determined by a behavioral assay and by electroretinographic (ERG) recording, zebrafish visual sensitivity was increased upon presentation of amino acids as olfactory stimuli. This effect, however, was observed only in the early morning hours when zebrafish are least sensitive to light. The effect of olfactory input on vision was eliminated after lesion of the olfactory bulbs or after the destruction of DA-IPCs. Intraocular injections of a dopamine D(2) but not a D(1) receptor antagonist blocked the effect of olfactory input on visual sensitivity. Although we cannot exclude the involvement of other anatomical pathways, our data suggest that the TN and DA-IPCs are the prime candidates for olfactory modulation of visual sensitivity. PMID- 12771170 TI - The V-type H(+)-ATPase in Malpighian tubules of Aedes aegypti: localization and activity. AB - The V-type H(+)-ATPase is thought to provide the driving force for transepithelial electrolyte and fluid secretion in Malpighian tubules. To confirm the presence of this proton pump in Malpighian tubules of the yellow fever mosquito Aedes aegypti, we used several antibodies raised against the V-type H(+) ATPase of Manduca sexta. Western blot analysis confirmed the presence of the V type H(+)-ATPase in Malpighian tubules of Aedes aegypti. In situ immunostaining identified the V-type H(+)-ATPase at the apical membrane of the mitochondrion rich brush border of principal cells. The V-type H(+)-ATPase was not found in stellate cells. Measurements of ATPase activity revealed that bafilomycin sensitive and NO(3)(-)-sensitive ATPase activity accounted for 50-60% of total ATPase activity in crude extracts of Malpighian tubules. No significant ouabain- or vanadate-sensitive Na(+)/K(+)-ATPase activity was detected. These results support the conclusion reached previously in electrophysiological studies that the mechanisms for transepithelial electrolyte secretion in the Aedes Malpighian tubules rely on the V-type H(+)-ATPase as the principal energizer of epithelial transport. Measures of transepithelial Na(+) and K(+) secretion and estimates of the H(+) flux mediated by the V-type H(+)-ATPase suggest a 1:1 stoichiometry for Na(+)/H(+) and K(+)/H(+) exchange transport across the apical membrane. PMID- 12771171 TI - Permeation and toxicity of ethylene glycol and methanol in larvae of Anopheles gambiae. AB - In this study, we applied proton NMR to measure the permeation of two cryoprotective agents (CPAs), ethylene glycol (EG) and methanol, into 1st instar Anopheles larvae. Calibration with standard solutions of EG or methanol (0-10 mol l(-1)) confirmed the reliability of the NMR measurements for determining the concentration of these solutes. To assess permeation, larvae were immersed in 1.5 mol l(-1) EG or 1.5 mol l(-1) methanol for different periods of time at 22 degrees C. The concentration of both CPAs in the larvae was then measured as a function of exposure time using (1)H-NMR spectroscopy. Results show that after a 6 h exposure to 1.5 mol l(-1) EG, the larval concentration of EG reaches a maximum value of 1.44 mol l(-1), which is 96% of the theoretical maximum. By contrast, after just 1 h exposure to 1.5 mol l(-1) methanol, the larval methanol concentration reaches its maximum, which, however, is only 75% of the theoretical maximum. Toxicity data show that larval survival remains 91% and 95% after 4 h and 1 h exposure to 1.5 mol l(-1) EG and 1.5 mol l(-1) methanol, respectively, at which time the larval concentration of EG and methanol has risen to 1.21 mol l ( 1) and 1.13 mol l(-1), respectively. These results suggest that CPAs such as EG and methanol do permeate Anopheles larvae to up to 81% and 75% of equilibrium, respectively, before the exposure becomes toxic. PMID- 12771172 TI - The representation of conspecific sounds in the auditory brainstem of teleost fishes. AB - Temporal patterns of sounds are thought to be the most important carriers of acoustic information in teleost fishes. In order to investigate how conspecific sounds are processed by the auditory system, auditory brainstem responses (ABRs) elicited by conspecific sounds were recorded in five species of teleosts. In the catfishes Platydoras costatus and Pimelodus pictus, the loach Botia modesta and the labyrinth fish Trichopsis vittata, all of which are hearing specialists, each pulse within the sounds elicited a separate brainwave that closely followed the temporal structure. The ABRs of P. costatus and B. modesta also represent amplitude patterns of conspecific sounds. By contrast, ABRs of the sunfish Lepomis gibbosus, a hearing non-specialist, consisted of long series of waves that could not be attributed to specific sound pulses. A more detailed analysis, however, indicated that each stimulus pulse contributed to the compound ABR waveform. Spectral analysis of low-pitched drumming sounds of P. pictus and corresponding ABRs showed peaks in the ABR spectra at the harmonics of the sound. Our results indicate that, besides temporal patterns, amplitude fluctuations and the frequency content of sounds can be represented in the auditory system and help the fish to extract important information for acoustic communication. PMID- 12771173 TI - Functional characterization of a glutamate/aspartate transporter from the mosquito Aedes aegypti. AB - Glutamate elicits a variety of effects in insects, including inhibitory and excitatory signals at both neuromuscular junctions and brain. Insect glutamatergic neurotransmission has been studied in great depth especially from the standpoint of the receptor-mediated effects, but the molecular mechanisms involved in the termination of the numerous glutamatergic signals have only recently begun to receive attention. In vertebrates, glutamatergic signals are terminated by Na(+)/K(+)-dependent high-affinity excitatory amino acid transporters (EAAT), which have been cloned and characterized extensively. Cloning and characterization of a few insect homologues have followed, but functional information for these homologues is still limited. Here we report a study conducted on a cloned mosquito EAAT homologue isolated from the vector of the dengue virus, Aedes aegypti. The deduced amino acid sequence of the protein, AeaEAAT, exhibits 40-50% identity with mammalian EAATs, and 45-50% identity to other insect EAATs characterized thus far. It transports L-glutamate as well as L and D-aspartate with high affinity in the micromolar range, and demonstrates a substrate-elicited anion conductance when heterologously expressed in Xenopus laevis oocytes, as found with mammalian homologues. Analysis of the spatial distribution of the protein demonstrates high expression levels in the adult thorax, which is mostly observed in the thoracic ganglia. Together, the work presented here provides a thorough examination of the role played by glutamate transport in Ae. aegypti. PMID- 12771174 TI - The influence of wing-wake interactions on the production of aerodynamic forces in flapping flight. AB - We used two-dimensional digital particle image velocimetry (DPIV) to visualize flow patterns around the flapping wing of a dynamically scaled robot for a series of reciprocating strokes starting from rest. The base of the wing was equipped with strain gauges so that the pattern of fluid motion could be directly compared with the time history of force production. The results show that the development and shedding of vortices throughout each stroke are highly stereotyped and influence force generation in subsequent strokes. When a wing starts from rest, it generates a transient force as the leading edge vortex (LEV) grows. This early peak, previously attributed to added-mass acceleration, is not amenable to quasi steady models but corresponds well to calculations based on the time derivative of the first moment of vorticity within a sectional slice of fluid. Forces decay to a stable level as the LEV reaches a constant size and remains attached throughout most of the stroke. The LEV grows as the wing supinates prior to stroke reversal, accompanied by an increase in total force. At stroke reversal, both the LEV and a rotational starting vortex (RSV) are shed into the wake, forming a counter-rotating pair that directs a jet of fluid towards the underside of the wing at the start of the next stroke. We isolated the aerodynamic influence of the wake by subtracting forces and flow fields generated in the first stroke, when the wake is just developing, from those produced during the fourth stroke, when the pattern of both the forces and wake dynamics has reached a limit cycle. This technique identified two effects of the wake on force production by the wing: an early augmentation followed by a small attenuation. The later decrease in force is consistent with the influence of a decreased aerodynamic angle of attack on translational forces caused by downwash within the wake and is well explained by a quasi-steady model. The early effect of the wake is not well approximated by a quasi-steady model, even when the magnitude and orientation of the instantaneous velocity field are taken into account. Thus, the wake capture force represents a truly unsteady phenomenon dependent on temporal changes in the distribution and magnitude of vorticity during stroke reversal. PMID- 12771175 TI - Regulation of branchial Na(+)/K(+)-ATPase in common carp Cyprinus carpio L. acclimated to different temperatures. AB - Isogenic carp Cyprinus carpio L. were acclimated to water temperatures of 15, 22 and 29 degrees C for at least 8 weeks. The acclimations consistently resulted in slightly, but significantly, different plasma osmolality, sodium, potassium and chloride concentrations between the groups studied. Plasma total and ionic calcium levels were unaffected, indicating successful adaptation. The apparent changes in set point for plasma ion levels are explained by altered sodium pump activity and hormonal control of branchial permeability to water and ions. It appears that in 15 degrees C-acclimated fish, a lower apparent Na(+)/K(+)-ATPase activity is compensated by strongly enhanced Na(+)/K(+)-ATPase expression (determined biochemically and immunohistochemically). In 29 degrees C-acclimated fish, the higher ambient temperature activates the enzyme. Arrhenius plots for branchial Na(+)/K(+)-ATPase preparations of the three groups of fish suggest the occurrence of different enzyme isoforms or protein (in)stability as explanations for differences in apparent enzyme activities, rather than temperature-dependent changes in membrane fluidity. As for hormonal control over permeability, prolactin mRNA expression (and anticipated production and release) is lower in fish kept at 29 degrees C, suggesting that control over branchial permeability to water and ions needs to be downregulated at higher temperatures. In so doing, enhanced sodium pump activity is balanced by a controlled passive ion loss to fine-tune plasma sodium levels. Basal plasma cortisol levels did not correlate positively with Na(+)/K(+)-ATPase expression, but doubling plasma cortisol levels in control fish by administering exogenous cortisol (for 7 days, using implanted minipumps and thus stress-free) enhanced Na(+)/K(+)-ATPase expression. This effect must be the result of a glucocorticoid action of the steroid: in fish, mineralocorticoid receptors have higher affinity for cortisol than glucocorticoid receptors. At a lower ambient temperature, branchial Na(+)/K(+)-ATPase expression is upregulated to counteract the temperature-inhibited activity of the sodium pump, perhaps via a mineralocorticoid receptor. PMID- 12771176 TI - Increase of internal ion concentration triggers trehalose synthesis associated with cryptobiosis in larvae of Polypedilum vanderplanki. AB - Larvae of an African chironomid, Polypedilum vanderplanki, which live in temporal rock pools, are completely dehydrated when the pools dry up and undergo anhydrobiosis until the next rain comes. During the dehydration process, larvae accumulate large amounts of trehalose, which provides effective protection against desiccation because of its high capacity for water replacement and vitrification. As the occurrence of rapid trehalose synthesis coincides with loss of body water, changes of osmolarity in the body are thought to be a cue for trehalose synthesis. Indeed, exposure to high salinity triggers rapid and efficient accumulation of trehalose even without desiccation treatment. As this rapid production occurs mainly in high concentrations of salt solutions, we conclude that an increase in internal ion concentration triggers trehalose synthesis associated with cryptobiosis in this species. PMID- 12771177 TI - Juvenile hormone and division of labor in honey bee colonies: effects of allatectomy on flight behavior and metabolism. AB - Three experiments were performed to determine why removal of the corpora allata (the glands that produce juvenile hormone) causes honey bees to fail to return to their hive upon initiating flight. In Experiment 1, the naturally occurring flights of allatectomized bees were tracked with radar to determine whether the deficit is physical or cognitive. The results indicated a physical impairment: allatectomized bees had a significantly slower ground speed than sham and untreated bees during orientation flights, but otherwise attributes such as flight range and area were normal. Flight impairment was confirmed in Experiment 2, based on observations of takeoff made in the field at the hive entrance. The allatectomized group had a significantly smaller percentage of flightworthy bees than did the sham and untreated groups. Experiment 3 confirmed the flight impairment in laboratory tests and showed that allatectomy causes a decrease in metabolic rate. Allatectomized bees had significantly lower metabolic rates than untreated and sham bees, while allatectomized bees receiving hormone replacement had intermediate values. These results indicate that allatectomy causes flight impairment, probably partly due to effects on metabolic rate. They also suggest that juvenile hormone plays an additional, previously unknown, role in coordinating the physiological underpinning of division of labor in honey bee colonies. PMID- 12771178 TI - Effect of anatomical distribution of mast cells on their defense function against bacterial infections: demonstration using partially mast cell-deficient tg/tg mice. AB - Mast cells were depleted in the peritoneal cavity of WBB6F1-tg/tg mice that did not express a transcription factor, MITF. When acute bacterial peritonitis was induced in WBB6F1-+/+, WBB6F1-W/Wv, and WBB6F1-tg/tg mice, the proportion of surviving WBB6F1-+/+ mice was significantly higher than that of surviving WBB6F1 W/Wv or WBB6F1-tg/tg mice. The poor survival of WBB6F1-W/Wv and WBB6F1-tg/tg mice was attributed to the deficient influx of neutrophils into the peritoneal cavity. The injection of cultured mast cells (CMCs) derived from WBB6F1-+/+ mice normalized the neutrophil influx and reduced survival rate in WBB6F1-W/Wv mice, but not in WBB6F1-tg/tg mice. This was not attributable to a defect of neutrophils because injection of TNF-alpha increased the neutrophil influx and survival rate in both WBB6F1-W/Wv and WBB6F1-tg/tg mice. Although WBB6F1-+/+ CMCs injection normalized the number of mast cells in both the peritoneal cavity and mesentery of WBB6F1-W/Wv mice, it normalized the number of mast cells only in the peritoneal cavity of WBB6F1-tg/tg mice. Mast cells within the mesentery or mast cells in the vicinity of blood vessels appeared to play an important role against the acute bacterial peritonitis. WBB6F1-tg/tg mice may be useful for studying the effect of anatomical distribution of mast cells on their antiseptic function. PMID- 12771179 TI - The role of intramolecular epitope spreading in the pathogenesis of endemic pemphigus foliaceus (fogo selvagem). AB - We report here a relationship between intramolecular epitope spreading and the clinical onset of the endemic form of pemphigus foliaceus in a Brazilian community with a high prevalence and incidence of the disease. Also known as Fogo Selvagem (FS), this disease is characterized by severe skin blistering and pathogenic anti-desmoglein-1 (Dsg1) autoantibodies. These autoantibodies bind the Dsg1 ectodomain and trigger keratinocyte cell detachment, the hallmark of FS. We show that (a) sera from FS patients in the preclinical stage recognized epitopes on the COOH-terminal EC5 domain of Dsg1, (b) disease onset was associated with the emergence of antibodies specific for epitopes on the NH2-terminal EC1 and EC2 domains, (c) all sera from FS patients with active disease recognized the EC1 and/or EC2 domains, and (d) sera from FS patients in remission showed reactivity restricted to EC5. These results suggest that anti-Dsg1 autoantibodies in FS are initially raised against the COOH-terminal EC5 domain of Dsg1 in individuals without skin disease; in genetically predisposed subjects the autoimmune response may then undergo intramolecular epitope spreading toward epitopes on the NH2 terminal EC1 and EC2 domains of Dsg1 leading to disease onset. Moreover, intramolecular epitope spreading may also modulate remissions and relapses of FS. PMID- 12771180 TI - Identification of a polymorphic gene, BCL2A1, encoding two novel hematopoietic lineage-specific minor histocompatibility antigens. AB - We report the identification of two novel minor histocompatibility antigens (mHAgs), encoded by two separate single nucleotide polymorphisms on a single gene, BCL2A1, and restricted by human histocompatibility leukocyte antigen (HLA) A*2402 (the most common HLA-A allele in Japanese) and B*4403, respectively. Two cytotoxic T lymphocyte (CTL) clones specific for these mHAgs were first isolated from two distinct recipients after hematopoietic cell transplantation. Both clones lyse only normal and malignant cells within the hematopoietic lineage. To localize the gene encoding the mHAgs, two-point linkage analysis was performed on the CTL lytic patterns of restricting HLA-transfected B lymphoblastoid cell lines obtained from Centre d'Etude du Polymorphisme Humain. Both CTL clones showed a completely identical lytic pattern for 4 pedigrees and the gene was localized within a 3.6-cM interval of 15q24.3-25.1 region that encodes at least 46 genes. Of those, only BCL2A1 has been reported to be expressed in hematopoietic cells and possess three nonsynonymous nucleotide changes. Minigene transfection and epitope reconstitution assays with synthetic peptides identified both HLA-A*2402- and B*4403-restricted mHAg epitopes to be encoded by distinct polymorphisms within BCL2A1. PMID- 12771183 TI - Protein translocation across the endoplasmic reticulum membrane in cold-adapted organisms. AB - Secretory proteins enter the secretory pathway by translocation across the membrane of the endoplasmic reticulum (ER) via a channel formed primarily by the Sec61 protein. Protein translocation is highly temperature dependent in mesophilic organisms. We asked whether the protein translocation machinery of organisms from extremely cold habitats was adapted to function at low temperature and found that post-translational protein import into ER-derived microsomes from Antarctic yeast at low temperature was indeed more efficient than into mesophilic yeast microsomes. Analysis of the amino-acid sequences of the core component of the protein translocation channel, Sec61p, from Antarctic yeast species did not reveal amino-acid changes potentially adaptive for function in the cold, because the sequences were too divergent. We therefore analyzed Sec61alpha (vertebrate Sec61p) sequences and protein translocation into the ER of Antarctic and Arctic fishes and compared them to Sec61alpha and protein translocation into the ER of temperate-water fishes and mammals. Overall, Sec61alpha is highly conserved amongst these divergent taxa; a number of amino-acid changes specific to fishes are evident throughout the protein, and, in addition, changes specific to cold water fishes cluster in the lumenal loop between transmembrane domains 7 and 8 of Sec61alpha, which is known to be important for protein translocation across the ER membrane. Secretory proteins translocated more efficiently into fish microsomes than into mammalian microsomes at 10 degrees C and 0 degrees C. The efficiency of protein translocation at 0 degrees C was highest for microsomes from a cold-water fish. Despite substantial differences in ER membrane lipid composition, ER membrane fluidity was identical in Antarctic fishes, mesophilic fishes and warm-blooded vertebrates, suggesting that membrane fluidity, although typically important for the function of the transmembrane proteins, is not limiting for protein translocation across the ER membrane in the cold. Collectively, our data suggest that the limited amino-acid changes in Sec61alpha from fishes may be functionally significant and represent adaptive changes that enhance channel function in the cold. PMID- 12771181 TI - Cbl-b negatively regulates B cell antigen receptor signaling in mature B cells through ubiquitination of the tyrosine kinase Syk. AB - Members of the Cbl family of molecular adaptors play key roles in regulating tyrosine kinase-dependent signaling in a variety of cellular systems. Here we provide evidence that in B cells Cbl-b functions as a negative regulator of B cell antigen receptor (BCR) signaling during the normal course of a response. In B cells from Cbl-b-deficient mice cross-linking the BCRs resulted in sustained phosphorylation of Igalpha, Syk, and phospholipase C (PLC)-gamma2, leading to prolonged Ca2+ mobilization, and increases in extracellular signal-regulated kinase (ERK) and c-Jun NH2-terminal protein kinase (JNK) phosphorylation and surface expression of the activation marker, CD69. Image analysis following BCR cross-linking showed sustained polarization of the BCRs into large signaling active caps associated with phosphorylated Syk in Cbl-b-deficient B cells in contrast to the BCRs in Cbl-b-expressing B cells that rapidly proceeded to form small, condensed, signaling inactive caps. Significantly, prolonged phosphorylation of Syk correlated with reduced ubiquitination of Syk indicating that Cbl-b negatively regulates BCR signaling by targeting Syk for ubiquitination. PMID- 12771182 TI - Inhibition of p38 MAPK signaling promotes late stages of myogenesis. AB - Signaling through the p38 mitogen-activated protein kinases (MAPKs) is essential for cartilage formation in primary cultures of limb mesenchyme. Here we show that, concurrent with a decrease in chondrogenesis, inhibition of p38 in limb bud cultures dramatically promotes muscle development. Specifically, treatment of primary limb bud cultures with p38 inhibitors increases the expression of myogenic markers and causes a striking increase in formation of myotubes, which were detected using antibodies specific for myosin heavy chain. These results are surprising in that they contrast with several previous reports describing a requirement for p38 during myogenesis. Nonetheless, the enhanced myogenesis leads to the formation of an extensive network of contractile myofibers, and this enhanced myogenesis can be conferred upon myogenic cells from clonal populations, such as G8 or C2C12 cells, if they are co-cultured with the limb mesenchymal cells. We provide evidence for the maintenance and rapid organization of existing, somitic-derived limb myoblasts in response to p38 inhibitors. These findings imply a novel and unexpected role for p38 MAPK inhibition in myogenesis and highlight the importance of the limb bud microenvironment in promoting the progression of limb myoblasts. PMID- 12771184 TI - Epidermal tissue regeneration and stromal interaction in HaCaT cells is initiated by TGF-alpha. AB - The human keratinocyte cell line HaCaT expresses essentially all epidermal differentiation markers but exhibits deficiencies in tissue organization as surface transplants in nude mice and even more so in organotypic co-cultures with fibroblasts. Whereas tissue differentiation by normal keratinocytes (NEKs) is regulated by stromal interactions, this mechanism is impaired in HaCaT cells. This regulatory process is initiated by interleukin-1 (IL-1) release in keratinocytes, which induces expression of keratinocyte growth factor (KGF/FGF-7) and granulocyte macrophage-colony stimulating factor (GM-CSF) in fibroblasts. Production and release of IL-1 is very low and, consequently, expression of the fibroblast-derived growth factors KGF/FGF-7 and GM-CSF is absent in HaCaT fibroblast co-cultures. However, addition of KGF and GMCSF, respectively, is inefficient to improve stratification and differentiation by HaCaT cells due to the low expression of their cognate receptors. More importantly, expression and release of the autocrine keratinocyte growth factor TGF-alpha is dramatically decreased in HaCaT cells. Addition of TGF- alpha or EGF stimulated HaCaT cell proliferation but, even more effectively, suppressed apoptosis, thus facilitating the formation of a regularly stratified epithelium. Furthermore, TGF-alpha enhanced the expression of the receptors for KGF and GM-CSF so that addition of these growth factors, or of their inducer IL-1, further improved epidermal tissue differentiation leading to in vitro skin equivalents comparable with cultures of NEKs. Thus, supplementing TGF-alpha normalized epidermal tissue regeneration by immortal HaCaT keratinocytes and their interaction with stromal cells so that regular skin equivalents are produced as standardized in vitro models. PMID- 12771185 TI - BTG2 antiproliferative protein interacts with the human CCR4 complex existing in vivo in three cell-cycle-regulated forms. AB - The yeast CCR4-NOT complex exists in two forms (1.0 and 1.9 MDa) that share several common subunits, including yCCR4, yCAF1 and five NOT proteins (NOT1-5). Here, we report that different complexes containing mammalian homologs of CCR4 NOT subunits exist in mammalian cells, with estimated sizes of approximately 1.9 MDa, approximately 1 MDa and approximately 650 kDa, and that BTG2, a member of a protein family with antiproliferative functions, can associate with these complexes. Immunoprecipitation and gel filtration experiments established that BTG2 interacts in vivo with hCCR4 protein via hCAF1 and hPOP2. Moreover, we show that hCCR4, as well as hCAF1 and BTG2, modulate the transcription regulation mediated by ERalpha. Finally, we demonstrate that the cellular localization of hCAF1 and the cell content in hCAF1-containing complexes change as cells progress from quiescence to S phase. These findings suggest that the different regulatory pathways in which hCAF1 is involved, notably transcription regulation and mRNA turnover, may occur through distinct CCR4 complexes in the course of cell-cycle progression. PMID- 12771186 TI - Phagosomal oxidative activity during beta2 integrin (CR3)-mediated phagocytosis by neutrophils is triggered by a non-restricted Ca2+ signal: Ca2+ controls time not space. AB - The temporal and spatial relationship between particle binding to the neutrophil by beta2 integrin (CR3), the Ca2+ elevation and subsequent oxidase activation has been unclear. This is because of the difficulty in studying the time course of individual phagocytic events in individual neutrophils. Here, we have used a micromanipulation technique to present C3bi-opsonised zymosan particles to the neutrophil under observation. In this way, the moment of particle contact, pseudopod formation and internalisation has been established and cytosolic free Ca2+ and oxidation of dichlorodihydrofluorescein (DCDHF)-labelled particles determined simultaneously. Using this approach, we have found that the Ca2+ signal, which is triggered by CR3-mediated phagocytosis, can be resolved into two temporally separated components. The first Ca2+ signal occurs during beta2 integrin engagement as the phagocytic cup forms but does not trigger oxidation of the particle. The second global Ca2+ signal, which is triggered about the time of phagosomal closure, causes an abrupt activation of the oxidase. This second Ca2+ signal was not restricted to the region of the phagosome yet only triggered the oxidase activation locally in the phagosome, with no evidence of activation at other sites in the neutrophil. This points to a dual control of oxidase activation, with Ca2+ controlling the timing of oxidase activation but slower and more localised molecular events, perhaps involving oxidase assembly and phosphatidylinositol 3-phosphate generation, determining the site of oxidase activation. PMID- 12771187 TI - The Crumbs3-Pals1 complex participates in the establishment of polarity in mammalian epithelial cells. AB - In Drosophila, the Crumbs-Stardust-Discs-lost complex is required during the establishment of polarized epithelia. Embryos that lack a component of this complex or overexpress Crumbs exhibit defects in epithelial morphogenesis. We recently cloned a novel mammalian epithelial Crumbs isoform, Crumbs3 (CRB3). CRB3 exists in a complex at tight junctions (TJs) with Pals1 and PATJ, the mammalian homologues of Stardust and Discs lost, respectively. Here, we observe that overexpression of CRB3 leads to delayed TJ formation in MDCK epithelial cell monolayers and disruption of polarity in MDCK cysts cultured in collagen. Both phenomena require the last four residues of CRB3. Next, we expressed, in MDCK cells, a dominant-negative Myc-Lin-2-Pals1 chimeric protein, where the PDZ domain of Lin-2 was replaced with that of Pals1. TJ and apical polarity defects were also observed in these cells. Collectively, this suggests that the CRB-Pals1 interaction is important for formation of TJs and polarized epithelia. These results provide insight into the function of the mammalian Crumbs complex during TJ formation and epithelial polarization. PMID- 12771188 TI - A STAT-regulated, stress-induced signalling pathway in Dictyostelium. AB - The Dictyostelium stalk cell inducer differentiation-inducing factor (DIF) directs tyrosine phosphorylation and nuclear accumulation of the STAT (signal transducer and activator of transcription) protein Dd-STATc. We show that hyperosmotic stress, heat shock and oxidative stress also activate Dd-STATc. Hyperosmotic stress is known to elevate intracellular cGMP and cAMP levels, and the membrane-permeant analogue 8-bromo-cGMP rapidly activates Dd-STATc, whereas 8 bromo-cAMP is a much less effective inducer. Surprisingly, however, Dd-STATc remains stress activatable in null mutants for components of the known cGMP mediated and cAMP-mediated stress-response pathways and in a double mutant affecting both pathways. Also, Dd-STATc null cells are not abnormally sensitive to hyperosmotic stress. Microarray analysis identified two genes, gapA and rtoA, that are induced by hyperosmotic stress. Osmotic stress induction of gapA and rtoA is entirely dependent on Dd-STATc. Neither gene is inducible by DIF but both are rapidly inducible with 8-bromo-cGMP. Again, 8-bromo-cAMP is a much less potent inducer than 8-bromo-cGMP. These data show that Dd-STATc functions as a transcriptional activator in a stress-response pathway and the pharmacological evidence, at least, is consistent with cGMP acting as a second messenger. PMID- 12771189 TI - Plastid ultrastructure defines the protein import pathway in dinoflagellates. AB - Eukaryotic cells contain a variety of different compartments that are distinguished by their own particular function and characteristic set of proteins. Protein targeting mechanisms to organelles have an additional layer of complexity in algae, where plastids may be surrounded by three or four membranes instead of two as in higher plants. The mechanism of protein import into dinoflagellates plastids, however, has not been previously described despite the importance of plastid targeting in a group of algae responsible for roughly half the ocean's net primary production. Here, we show how nuclear-encoded proteins enter the triple membrane-bound plastids of the dinoflagellate Gonyaulax. These proteins all contain an N-terminal leader sequence with two distinct hydrophobic regions flanking a region rich in hydroxylated amino acids (S/T). We demonstrate that plastid proteins transit through the Golgi in vivo, that the first hydrophobic region in the leader acts as a typical signal peptide in vitro, and that the S/T-rich region acts as a typical plastid transit sequence in transgenic plants. We also show that the second hydrophobic region acts as a stop transfer sequence so that plastid proteins in Golgi-derived vesicles are integral membrane proteins with a predominant cytoplasmic component. The dinoflagellate mechanism is thus different from that used by the phylogenetically related apicomplexans, and instead, is similar to that of the phylogenetically distant Euglena, whose plastids are also bound by three membranes. We conclude that the protein import mechanism is dictated by plastid ultrastructure rather than by the evolutionary history of the cell. PMID- 12771190 TI - SETA/CIN85/Ruk and its binding partner AIP1 associate with diverse cytoskeletal elements, including FAKs, and modulate cell adhesion. AB - The adaptor protein SETA/CIN85/Ruk is involved in regulating diverse signal transduction pathways, including the internalization of tyrosine kinase receptors via the Cbl ubiquitin ligases, and attenuating PI3K activity by interaction with its regulatory subunit. Here we present evidence for a new aspect of SETA function, based on the initial observation that it co-localizes with actin in microfilaments and at focal adhesions, and with microtubules. Although there was no evidence for direct molecular interactions between SETA and cytoskeletal proteins, the SETA-interacting protein AIP1, which is a rat ortholog of the Xenopus src substrate Xp95, strongly interacted with structural proteins of the cytoskeleton, including actin and tubulins. Both SETA and AIP1 interacted with focal adhesion kinase (FAK) and proline rich tyrosine kinase 2 (PYK-2), and c-Cbl interacted with PYK-2. AIP1, which interacted more strongly than either SETA or c Cbl, required an intact consensus tyrosine kinase phosphorylation sequence at Y319 to bind to focal adhesion kinases, which suggests that phosphorylation is an important mediator of this complex. SETA, which interacted as a dimer with focal adhesion kinases, promoted the interaction between PYK-2 and AIP1. Direct analysis of the impact of these proteins on cell adhesion, by use of an electrical cell-substrate impedance sensor (ECIS), showed that SETA promoted cell adhesion while AIP1 and c-Cbl reduced it. Furthermore, the ability of AIP1 and AIP1 mutants to decrease cell adhesion in ECIS analysis correlated with their presence in PYK-2 complexes, providing a direct link between AIP1-mediated molecular interactions and cellular behavior. Transfection of AIP1 also reduced the level of phosphorylation of endogenous PYK-2 and FAK, suggesting that this protein may directly regulate focal adhesion kinases, and thereby cell adhesion. These data are the first to implicate the adaptor protein SETA and its binding partner AIP1 as being involved with the cytoskeleton and in the regulation of cell adhesion, and suggest that they may be part of the focal adhesion kinase regulatory complex. PMID- 12771191 TI - Custom distinctions in the interaction of G-protein beta subunits with N-type (CaV2.2) versus P/Q-type (CaV2.1) calcium channels. AB - Inhibition of N- (Cav2.2) and P/Q-type (Cav2.1) calcium channels by G-proteins contribute importantly to presynaptic inhibition as well as to the effects of opiates and cannabinoids. Accordingly, elucidating the molecular mechanisms underlying G-protein inhibition of voltage-gated calcium channels has been a major research focus. So far, inhibition is thought to result from the interaction of multiple proposed sites with the Gbetagamma complex (Gbetagamma). Far less is known about the important interaction sites on Gbetagamma itself. Here, we developed a novel electrophysiological paradigm, "compound-state willing reluctant analysis," to describe Gbetagamma interaction with N- and P/Q-type channels, and to provide a sensitive and efficient screen for changes in modulatory behavior over a broad range of potentials. The analysis confirmed that the apparent (un)binding kinetics of Gbetagamma with N-type are twofold slower than with P/Q-type at the voltage extremes, and emphasized that the kinetic discrepancy increases up to ten-fold in the mid-voltage range. To further investigate apparent differences in modulatory behavior, we screened both channels for the effects of single point alanine mutations within four regions of Gbeta1, at residues known to interact with Galpha. These residues might thereby be expected to interact with channel effectors. Of eight mutations studied, six affected G-protein modulation of both N- and P/Q-type channels to varying degrees, and one had no appreciable effect on either channel. The remaining mutation was remarkable for selective attenuation of effects on P/Q-, but not N type channels. Surprisingly, this mutation decreased the (un)binding rates without affecting its overall affinity. The latter mutation suggests that the binding surface on Gbetagamma for N- and P/Q-type channels are different. Also, the manner in which this last mutation affected P/Q-type channels suggests that some residues may be important for "steering" or guiding the protein into the binding pocket, whereas others are important for simply binding to the channel. PMID- 12771192 TI - A cysteine scan of the inner vestibule of cyclic nucleotide-gated channels reveals architecture and rearrangement of the pore. AB - Cyclic nucleotide-gated (CNG) channels belong to the P-loop-containing family of ion channels that also includes KcsA, MthK, and Shaker channels. In this study, we investigated the structure and rearrangement of the CNGA1 channel pore using cysteine mutations and cysteine-specific modification. We constructed 16 mutant channels, each one containing a cysteine mutation at one of the positions between 384 and 399 in the S6 region of the pore. By measuring currents activated by saturating concentrations of the full agonist cGMP and the partial agonists cIMP and cAMP, we show that mutating S6 residues to cysteine caused both favorable and unfavorable changes in the free energy of channel opening. The time course of cysteine modification with 2-aminoethylmethane thiosulfonate hydrochloride (MTSEA) was complex. For many positions we observed decreases in current activated by cGMP and concomitant increases in current activated by cIMP and cAMP. A model where modification affected both gating and permeation successfully reproduced the complex time course of modification for most of the mutant channels. From the model fits to the time course of modification for each mutant channel, we quantified the following: (a) the bimolecular rate constant of modification in the open state, (b) the change in conductance, and (c) the change in the free energy of channel opening for modification of each cysteine. At many S6 cysteines, modification by MTSEA caused a decrease in conductance and a favorable change in the free energy of channel opening. Our results are interpreted within the structural framework of the known structures of KcsA and MthK. We conclude that: (a) MTSEA modification affects both gating and permeation, (b) the open configuration of the pore of CNGA1 channels is consistent with the structure of MthK, and (c) the modification of S6 residues disrupts the helical packing of the closed channel, making it easier for channels to open. PMID- 12771193 TI - Saxitoxin is a gating modifier of HERG K+ channels. AB - Potassium (K+) channels mediate numerous electrical events in excitable cells, including cellular membrane potential repolarization. The hERG K+ channel plays an important role in myocardial repolarization, and inhibition of these K+ channels is associated with long QT syndromes that can cause fatal cardiac arrhythmias. In this study, we identify saxitoxin (STX) as a hERG channel modifier and investigate the mechanism using heterologous expression of the recombinant channel in HEK293 cells. In the presence of STX, channels opened slower during strong depolarizations, and they closed much faster upon repolarization, suggesting that toxin-bound channels can still open but are modified, and that STX does not simply block the ion conduction pore. STX decreased hERG K+ currents by stabilizing closed channel states visualized as shifts in the voltage dependence of channel opening to more depolarized membrane potentials. The concentration dependence for steady-state modification as well as the kinetics of onset and recovery indicate that multiple STX molecules bind to the channel. Rapid application of STX revealed an apparent "agonist-like" effect in which K+ currents were transiently increased. The mechanism of this effect was found to be an effect on the channel voltage-inactivation relationship. Because the kinetics of inactivation are rapid relative to activation for this channel, the increase in K+ current appeared quickly and could be subverted by a decrease in K+ currents due to the shift in the voltage-activation relationship at some membrane potentials. The results are consistent with a simple model in which STX binds to the hERG K+ channel at multiple sites and alters the energetics of channel gating by shifting both the voltage-inactivation and voltage-activation processes. The results suggest a novel extracellular mechanism for pharmacological manipulation of this channel through allosteric coupling to channel gating. PMID- 12771194 TI - Negative charges in the transmembrane domains of the HERG K channel are involved in the activation- and deactivation-gating processes. AB - The transmembrane domains of HERG (S1-S3) contain six negative charges: three are conserved in all voltage-gated K channels (D456 and D466 in S2, D501 in S3) and three are unique to the EAG family (D411 in S1, D460 in S2, and D509 in S3). We infer the functional role of these aspartates by studying how substituting them with cysteine, one at a time, affects the channel function. D456C is not functional, suggesting that this negative charge may play a critical role in channel protein folding during biogenesis, as has been shown for its counterpart in the Shaker channel. Data from the other five functional mutants suggest that D411 can stabilize the HERG channel in the closed state, while D460 and D509 have the opposite effect. D466 and D501 both may contribute to voltage-sensing during the activation process. On the other hand, all five aspartates work in a concerted fashion in contributing to the slow deactivation process of the HERG channel. Accessibility tests of the introduced thiol groups to extracellular MTS reagents indicate that water-filled crevices penetrate deep into the HERG protein core, reaching the cytoplasmic halves of S1 and S2. At these deep locations, accessibility of 411C and 466C to the extracellular aqueous phase is voltage dependent, suggesting that conformational changes occur in S1 and S2 or the surrounding crevices during gating. Increasing extracellular [H+] accelerates HERG deactivation. This effect is suppressed by substituting the aspartates with cysteine, suggesting that protonation of these aspartates may contribute to the signaling pathway whereby external [H+] influences conformational changes in the channel's cytoplasmic domains (where deactivation takes place). There is no evidence for a metal ion binding site coordinated by negative charges in the transmembrane domains of HERG, as the one described for the EAG channel. PMID- 12771195 TI - Properties of single voltage-gated proton channels in human eosinophils estimated by noise analysis and by direct measurement. AB - Voltage-gated proton channels were studied under voltage clamp in excised, inside out patches of human eosinophils, at various pHi with pHo 7.5 or 6.5 pipette solutions. H+ current fluctuations were observed consistently when the membrane was depolarized to voltages that activated H+ current. At pHi < or = 5.5 the variance increased nonmonotonically with depolarization to a maximum near the midpoint of the H+ conductance-voltage relationship, gH-V, and then decreased, supporting the idea that the noise is generated by H+ channel gating. Power spectral analysis indicated Lorentzian and 1/f components, both related to H+ currents. Unitary H+ current amplitude was estimated from stationary or quasi stationary variance, sigmaH2. We analyze sigmaH2 data obtained at various voltages on a linearized plot that provides estimates of both unitary conductance and the number of channels in the patch, without requiring knowledge of open probability. The unitary conductance averaged 38 fS at pHi 6.5, and increased nearly fourfold to 140 fS at pHi 5.5, but was independent of pHo. In contrast, the macroscopic gH was only 1.8-fold larger at pHi 5.5 than at pHi 6.5. The maximum H+ channel open probability during large depolarizations was 0.75 at pHi 6.5 and 0.95 at pHi 5.5. Because the unitary conductance increases at lower pHi more than the macroscopic gH, the number of functional channels must decrease. Single H+ channel currents were too small to record directly at physiological pH, but at pHi < or = 5.5 near Vthreshold (the voltage at which gH turns on), single channel-like current events were observed with amplitudes 7-16 fA. PMID- 12771196 TI - Structural variations and stabilising modifications of synthetic siRNAs in mammalian cells. AB - Double-stranded short interfering RNAs (siRNA) induce post-transcriptional silencing in a variety of biological systems. In the present study we have investigated the structural requirements of chemically synthesised siRNAs to mediate efficient gene silencing in mammalian cells. In contrast to studies with Drosophila extracts, we found that synthetic, double-stranded siRNAs without specific nucleotide overhangs are highly efficient in gene silencing. Blocking of the 5'-hydroxyl terminus of the antisense strand leads to a dramatic loss of RNA interference activity, whereas blocking of the 3' terminus or blocking of the termini of the sense strand had no negative effect. We further demonstrate that synthetic siRNA molecules with internal 2'-O-methyl modification, but not molecules with terminal modifications, are protected against serum-derived nucleases. Finally, we analysed different sets of siRNA molecules with various 2' O-methyl modifications for stability and activity. We demonstrate that 2'-O methyl modifications at specific positions in the molecule improve stability of siRNAs in serum and are tolerated without significant loss of RNA interference activity. These second generation siRNAs will be better suited for potential therapeutic application of synthetic siRNAs in vivo. PMID- 12771197 TI - Insight into the mechanism of the peptide-based gene delivery system MPG: implications for delivery of siRNA into mammalian cells. AB - The improvement of non-viral-based gene delivery systems is of prime importance for the future of gene and antisense therapies. We have previously described a peptide-based gene delivery system, MPG, derived from the fusion peptide domain of HIV-1 gp41 protein and the nuclear localisation sequence (NLS) of SV40 large T antigen. MPG forms stable non-covalent complexes with nucleic acids and improves their delivery. In the present work, we have investigated the mechanism through which MPG promotes gene delivery. We demonstrate that cell entry is independent of the endosomal pathway and that the NLS of MPG is involved in both electrostatic interactions with DNA and nuclear targeting. MPG/DNA particles interact with the nuclear import machinery, however, a mutation which affects the NLS of MPG disrupts these interactions and prevents nuclear delivery of DNA. Nevertheless, we show that this mutation yields a variant of MPG which is a powerful tool for delivery of siRNA into mammalian cells, enabling rapid release of the siRNA into the cytoplasm and promoting robust down-regulation of target mRNA. Taken together, these results support the potential of MPG-like peptides for therapeutic applications and suggest that specific variations in the sequence may yield carriers with distinct targeting features. PMID- 12771198 TI - Syntheses and structural studies of calix[4]arene-nucleoside and calix[4]arene oligonucleotide hybrids. AB - We have synthesized three types of calix[4]arene- nucleoside hybrid efficiently by amide bond formation between the amine functional groups of 1,3 diaminocalix[4]arene and the carboxyl groups of thymidine nucleoside derivatives. X-ray crystallography of a homocoupled calix[4]arene-nucleoside hybrid revealed an interesting hydrogen bonding pattern between thymine bases and the amide linkages. We designed the calix[4]arene-oligonucleotide hybrids (5' AAAAGATATCAAXTTGATATCTTTT-3', 5'-T12-X-T12-3', and 5'-A12-X-T12-3') to be V shaped oligodeoxyribonucleotides and synthesized them by using a calix[4]arene nucleoside hybrid (X) as a key building block. Thermal denaturation experiments, monitored by UV spectroscopy at 260 and 284 nm, and circular dichroism spectra of the calix[4]arene-oligonucleotide hybrids suggest that the modified oligonucleotides indeed adopt V-shaped conformations, making them suitable for use as building blocks in the construction of programmed oligonucleotide nanostructures PMID- 12771199 TI - Transcriptional regulation of the human MIP-1alpha promoter by RUNX1 and MOZ. AB - The transcription factor RUNX1 (AML-1, PEBP2alphaB and CBFA2) is essential for definitive haematopoiesis, and chromosomal translocations involving the RUNX1 gene are frequently found in acute leukaemias. The gene encoding the histone acetyltransferase MOZ is also rearranged in some acute leukaemias, resulting in the expression of MOZ fusion proteins. MOZ has recently been shown to interact directly with RUNX1, indicating that MOZ fusion proteins act by deregulating RUNX1 function. Macrophage inflammatory protein-1alpha (MIP-1alpha) is a proinflammatory cytokine that also inhibits proliferation of haematopoietic stem cells. Amongst the conserved sequence elements in the human MIP-1alpha promoter are two consensus RUNX sites. We have investigated the role of these RUNX sites in the regulation of the MIP-1alpha promoter by PMA/PHA stimulation in Jurkat T cells. RUNX1 can specifically bind to both RUNX sites in vitro and chromatin immunoprecipitation assays demonstrated that endogenous RUNX1 is constitutively bound to the endogenous MIP-1alpha promoter. Mutation of the RUNX sites demonstrated that the proximal RUNX site is essential for PMA/PHA-stimulated activation of the MIP-1alpha promoter. Activation of the promoter can also be inhibited by heterologous expression of the repressor protein AML-1/ETO. We further demonstrate that MOZ can activate the MIP-1alpha promoter and that this activation is largely dependent upon the proximal RUNX site. Moreover, we show that co-expression of MOZ and RUNX1 can synergistically activate the MIP-1alpha promoter. The regulation of MIP-1alpha expression by RUNX1/MOZ is discussed in the context of MIP-1alpha's role as an inhibitor of haematopoietic stem cell proliferation and its potential importance in leukaemias associated with RUNX1 or MOZ chromosomal rearrangements. PMID- 12771200 TI - Effect of DNA bases and backbone on sigma70 holoenzyme binding and isomerization using fork junction probes. AB - Abasic substitutions in the non-template strand and promoter sequence changes were made to assess the roles of various promoter features in sigma70 holoenzyme interactions with fork junction probes. Removal of -10 element non-template single strand bases, leaving the phosphodiester backbone intact, did not interfere with binding. In contrast these abasic probes were deficient in promoting holoenzyme isomerization to the heparin resistant conformation. Thus, it appears that the melted -10 region interaction has two components, an initial enzyme binding primarily to the phosphodiester backbone and a base dependent isomerization of the bound enzyme. In contrast various upstream elements cooperate primarily to stimulate binding. Features and positions most important for these effects are identified. PMID- 12771201 TI - The phage T4 restriction endoribonuclease RegB: a cyclizing enzyme that requires two histidines to be fully active. AB - The regB gene, from the bacteriophage T4, codes for an endoribonuclease that controls the expression of a number of phage early genes. The RegB protein cleaves its mRNA substrates with an almost absolute specificity in the middle of the tertranucleotide GGAG, making it a unique well-defined restriction endoribonuclease. This striking protein has no homology to any known RNase and its catalytic mechanism has never been investigated. Here, we show, using 31P nuclear magnetic resonance (NMR), that RegB produces a cyclic 2',3' phosphodiester product. In order to determine the residues crucial for its activity, we prepared all the histidine-to- alanine point mutants of RegB. The activity of these mutants was characterized both in vivo and in vitro. In addition, their binding capability was quantified by surface plasmon resonance and their structural integrity was probed by 1H/15N NMR correlation spectroscopy. The results obtained show that only the H48A and the H68A substitutions significantly reduce RegB activity without changing its ability to bind the substrate or affecting its overall structure. Altogether, our results define RegB as a new cyclizing RNase and present His48 and His68 as potent catalytic residues. The effect of the in vivo selected R52L mutation is also described and discussed. PMID- 12771202 TI - Steric inhibition of human immunodeficiency virus type-1 Tat-dependent trans activation in vitro and in cells by oligonucleotides containing 2'-O-methyl G clamp ribonucleoside analogues. AB - We report the synthesis of a novel 2'-O-methyl (OMe) riboside phosphoramidite derivative of the G-clamp tricyclic base and incorporation into a series of small steric blocking OMe oligonucleotides targeting the apical stem-loop region of human immunodeficiency virus type 1 (HIV-1) trans- activation-responsive (TAR) RNA. Binding to TAR RNA is substantially enhanced for certain single site substitutions in the centre of the oligonucleotide, and doubly substituted anti TAR OMe 9mers or 12mers exhibit remarkably low binding constants of <0.1 nM. G clamp-containing oligomers achieved 50% inhibition of Tat-dependent in vitro transcription at approximately 25 nM, 4-fold lower than for a TAR 12mer OMe oligonucleotide and better than found for any other oligonucleotide tested to date. Addition of one or two OMe G-clamps did not impart cellular trans activation inhibition activity to cellularly inactive OMe oligonucleotides. Addition of an OMe G-clamp to a 12mer OMe-locked nucleic acid chimera maintained, but did not enhance, inhibition of Tat-dependent in vitro transcription and cellular trans-activation in HeLa cells. The results demonstrate clearly that an OMe G-clamp has remarkable RNA-binding enhancement ability, but that oligonucleotide effectiveness in steric block inhibition of Tat-dependent trans activation both in vitro and in cells is governed by factors more complex than RNA-binding strength alone. PMID- 12771203 TI - Overexpression of FABP7 in Down syndrome fetal brains is associated with PKNOX1 gene-dosage imbalance. AB - Suppression subtractive hybridization performed on Down syndrome (DS) fetal brains revealed a differentially expressed gene, FABP7, mapped to 6q22-23. FABP7 overexpression in DS brains was verified by real-time PCR (1.63-fold). To elucidate the molecular basis of FABP7 overexpression and establish the relationship with chromosome 21 trisomy, the FABP7 promoter was cloned by genomic inverse PCR. Comparison to the mouse ortholog revealed conservation of reported regulatory elements, among them a Pbx/POU binding site, known to be the target of PBX heteromeric complexes. PBX partners include homeobox-containing proteins, such as PKNOX1 (PREP1), a transcription factor mapping at 21q22.3. We report here: (i) overexpression of PKNOX1 in DS fetal brains; (ii) in vitro specific binding of PKNOX1 to the Pbx/POU site of the FABP7 promoter; (iii) in vivo FABP7 promoter trans-activation in cultured neuroblastoma cells caused by PKNOX1 overexpression. To our knowledge this is the first report of a direct relation between dosage imbalance of a chromosome 21 gene and altered expression of a downstream gene mapping on another chromosome. Given the role of FABP7 in the establishment, development and maintenance of the CNS, we suggest that the overexpression of FABP7 could contribute to DS-associated neurological disorders. PMID- 12771204 TI - Domain mapping of Escherichia coli RecQ defines the roles of conserved N- and C terminal regions in the RecQ family. AB - RecQ DNA helicases function in DNA replication, recombination and repair. Although the precise cellular roles played by this family of enzymes remain elusive, the importance of RecQ proteins is clear; mutations in any of three human RecQ genes lead to genomic instability and cancer. In this report, proteolysis is used to define a two-domain structure for Escherichia coli RecQ, revealing a large (approximately 59 kDa) N-terminal and a small (approximately 9 kDa) C-terminal domain. A short N-terminal segment (7 or 21 residues) is also shown to be sensitive to proteases. The effects of removing these regions of RecQ are tested in vitro. Removing 21 N-terminal residues from RecQ severely diminishes its DNA-dependent ATPase and helicase activities, but does not affect its ability to bind DNA in electrophoretic mobility shift assays. In contrast, removing the approximately 9 kDa C-terminal domain from RecQ results in a fragment with normal levels of ATPase and helicase activity, but that has lost the ability to stably associate with DNA. These results establish the biochemical roles of an N-terminal sequence motif in RecQ catalytic function and for the C terminal RecQ domain in stable DNA binding. PMID- 12771205 TI - UVA-induced cyclobutane pyrimidine dimers form predominantly at thymine-thymine dipyrimidines and correlate with the mutation spectrum in rodent cells. AB - Ligation-mediated PCR was employed to quantify cyclobutane pyrimidine dimer (CPD) formation at nucleotide resolution along exon 2 of the adenine phosphoribosyltransferase (aprt) locus in Chinese hamster ovary (CHO) cells following irradiation with either UVA (340-400 nm), UVB (295-320 nm), UVC (254 nm) or simulated sunlight (SSL; lambda > 295 nm). The resulting DNA damage spectrum for each wavelength region was then aligned with the corresponding mutational spectrum generated previously in the same genetic target. The DNA sequence specificities of CPD formation induced by UVC, UVB or SSL were very similar, i.e., in each case the overall relative proportion of this photoproduct forming at TT, TC, CT and CC sites was approximately 28, approximately 26, approximately 16 and approximately 30%, respectively. Furthermore, a clear correspondence was noted between the precise locations of CPD damage hotspots, and of 'UV signature' mutational hotspots consisting primarily of C-->T and CC- >TT transitions within pyrimidine runs. However, following UVA exposure, in strong contrast to the above situation for UVC, UVB or SSL, CPDs were generated much more frequently at TT sites than at TC, CT or CC sites (57% versus 18, 11 and 14%, respectively). This CPD deposition pattern correlates well with the strikingly high proportion of mutations recovered opposite TT dipyrimidines in UVA- irradiated CHO cells. Our results directly implicate the CPD as a major promutagenic DNA photoproduct induced specifically by UVA in rodent cells. PMID- 12771206 TI - Faithful expression of a tagged Fugu WT1 protein from a genomic transgene in zebrafish: efficient splicing of pufferfish genes in zebrafish but not mice. AB - The teleost fish are widely used as model organisms in vertebrate biology. The compact genome of the pufferfish, Fugu rubripes, has proven a valuable tool in comparative genome analyses, aiding the annotation of mammalian genomes and the identification of conserved regulatory elements, whilst the zebrafish is particularly suited to genetic and developmental studies. We demonstrate that a pufferfish WT1 transgene can be expressed and spliced appropriately in transgenic zebrafish, contrasting with the situation in transgenic mice. By creating both transgenic mice and transgenic zebrafish with the same construct, we show that Fugu RNA is processed correctly in zebrafish but not in mice. Furthermore, we show for the first time that a Fugu genomic construct can produce protein in transgenic zebrafish: a full-length Fugu WT1 transgene with a C-terminal beta galactosidase fusion is spliced and translated correctly in zebrafish, mimicking the expression of the endogenous WT1 gene. These data demonstrate that the zebrafish:Fugu system is a powerful and convenient tool for dissecting both vertebrate gene regulation and gene function in vivo. PMID- 12771207 TI - Purification and characterisation of a novel DNA methyltransferase, M.AhdI. AB - We have cloned the M and S genes of the restriction-modification (R-M) system AhdI and have purified the resulting methyltransferase to homogeneity. M.AhdI is found to form a 170 kDa tetrameric enzyme having a subunit stoichiometry M2S2 (where the M and S subunits are responsible for methylation and DNA sequence specificity, respectively). Sedimentation equilibrium experiments show that the tetrameric enzyme dissociates to form a heterodimer at low concentration, with K(d) approximately 2 microM. The intact (tetrameric) enzyme binds specifically to a 30 bp DNA duplex containing the AhdI recognition sequence GACN5GTC with high affinity (K(d) approximately 50 nM), but at low enzyme concentration the DNA binding activity is governed by the dissociation of the tetramer into dimers, leading to a sigmoidal DNA binding curve. In contrast, only non-specific binding is observed if the duplex lacks the recognition sequence. Methylation activity of the purified enzyme was assessed by its ability to prevent restriction by the cognate endonuclease. The subunit structure of the M.AhdI methyltransferase resembles that of type I MTases, in contrast to the R.AhdI endonuclease which is typical of type II systems. AhdI appears to be a novel R-M system with properties intermediate between simple type II systems and more complex type I systems, and may represent an intermediate in the evolution of R-M systems. PMID- 12771208 TI - Using structural motif templates to identify proteins with DNA binding function. AB - This work describes a method for predicting DNA binding function from structure using 3-dimensional templates. Proteins that bind DNA using small contiguous helix-turn-helix (HTH) motifs comprise a significant number of all DNA-binding proteins. A structural template library of seven HTH motifs has been created from non-homologous DNA-binding proteins in the Protein Data Bank. The templates were used to scan complete protein structures using an algorithm that calculated the root mean squared deviation (rmsd) for the optimal superposition of each template on each structure, based on C(alpha) backbone coordinates. Distributions of rmsd values for known HTH-containing proteins (true hits) and non-HTH proteins (false hits) were calculated. A threshold value of 1.6 A rmsd was selected that gave a true hit rate of 88.4% and a false positive rate of 0.7%. The false positive rate was further reduced to 0.5% by introducing an accessible surface area threshold value of 990 A2 per HTH motif. The template library and the validated thresholds were used to make predictions for target proteins from a structural genomics project. PMID- 12771209 TI - Solution structure of the influenza A virus cRNA promoter: implications for differential recognition of viral promoter structures by RNA-dependent RNA polymerase. AB - Influenza A virus replication requires the interaction of viral RNA-dependent RNA polymerase (RdRp) with promoters in both the RNA genome (vRNA) and the full length complementary RNA (cRNA) which serve as templates for the generation of new vRNAs. Although RdRp binds both promoters effectively, it must also discriminate between them because they serve different functional roles in the viral life cycle. Even though the inherent asymmetry between two RNA promoters is considered as a cause of the differential recognition by the RdRp, the structural basis for the ability of the RdRp to recognize the RNA promoters and discriminate effectively between them remains unsolved. Here we report the structure of the cRNA promoter of influenza A virus as determined by heteronuclear magnetic resonance spectroscopy. The terminal region is extremely unstable and does not have a rigid structure. The major groove of the internal loop is widened by the displacement of a novel A*(UU) motif toward the minor groove. These internal loop residues show distinguishable dynamic characters, with differing motional timescales for each residue. Comparison of the cRNA promoter structure with that of the vRNA promoter reveals common structural and dynamic elements in the internal loop, but also differences that provide insight into how the viral RdRp differentially recognizes the cRNA and vRNA promoters. PMID- 12771210 TI - SPINE 2: a system for collaborative structural proteomics within a federated database framework. AB - We present version 2 of the SPINE system for structural proteomics. SPINE is available over the web at http://nesg.org. It serves as the central hub for the Northeast Structural Genomics Consortium, allowing collaborative structural proteomics to be carried out in a distributed fashion. The core of SPINE is a laboratory information management system (LIMS) for key bits of information related to the progress of the consortium in cloning, expressing and purifying proteins and then solving their structures by NMR or X-ray crystallography. Originally, SPINE focused on tracking constructs, but, in its current form, it is able to track target sample tubes and store detailed sample histories. The core database comprises a set of standard relational tables and a data dictionary that form an initial ontology for proteomic properties and provide a framework for large-scale data mining. Moreover, SPINE sits at the center of a federation of interoperable information resources. These can be divided into (i) local resources closely coupled with SPINE that enable it to handle less standardized information (e.g. integrated mailing and publication lists), (ii) other information resources in the NESG consortium that are inter-linked with SPINE (e.g. crystallization LIMS local to particular laboratories) and (iii) international archival resources that SPINE links to and passes on information to (e.g. TargetDB at the PDB). PMID- 12771211 TI - Integrated functional and bioinformatics approach for the identification and experimental verification of RNA signals: application to HIV-1 INS. AB - Regulation of gene expression involves sequence elements in nucleic acids. In promoters, multiple sequence elements cooperate as functional modules, which in combination determine overall promoter activity. We previously developed computational tools based on this hierarchical structure for in silico promoter analysis. Here we address the functional organization of post-transcriptional control elements, using the HIV-1 genome as a model. Numerous mutagenesis studies demonstrate that expression of HIV structural proteins is restricted by inhibitory sequences (INS) in HIV mRNAs in the absence of the HIV-1 Rev protein. However, previous attempts to detect conserved sequence patterns of HIV-1 INS have failed. We defined four distinct sequence patterns for inhibitory motifs (weight matrices), which identified 22 out of the 25 known INS as well as several new candidate INS regions contained in numerous HIV-1 strains. The conservation of INS motifs within the HIV genome was not due to overall sequence conservation. The functionality of two candidate INS regions was analyzed with a new assay that measures the effect of non-coding mRNA sequences on production of red fluorescent reporter protein. Both new INS regions showed inhibitory activity in sense but not in antisense orientation. Inhibitory activity increased by combining both INS regions in the same mRNA. Inhibitory activity of known and new INS regions was overcome by co-expression of the HIV-1 Rev protein. PMID- 12771213 TI - Tertiary structure base pairs between D- and TpsiC-loops of Escherichia coli tRNA(Leu) play important roles in both aminoacylation and editing. AB - To ensure the fidelity of protein biosynthesis, aminoacyl-tRNA synthetases (aaRSs) must recognize the tRNA identity elements of their cognate tRNAs and discriminate their cognate amino acids from structurally similar ones through a proofreading (editing) reaction. For a better understanding of these processes, we investigated the role of tRNA(Leu) tertiary structure in the aminoacylation and editing reactions catalyzed by leucyl-tRNA synthetase (LeuRS). We constructed a series of Escherichia coli tRNA(Leu) mutated transcripts with alterations of the nucleotides involved in tertiary interactions. Our results revealed that any disturbance of the tertiary interaction between the tRNA(Leu) D- and TpsiC-loops affected both its aminoacylation ability and its ability to stimulate the editing reaction. Moreover, we found that the various tertiary interactions between the D and TpsiC-loops (G18:U55, G19:C56 and U54:A58) functioned differently within the aminoacylation and editing reactions. In these two reactions, the role of base pair 19:56 was closely correlated and dependent on the hydrogen bond number. In contrast, U54:A58 was more important in aminoacylation than in editing. Taken together, our results suggest that the elbow region of tRNA formed by the tertiary interactions between the D- and TpsiC-loops affects the interactions between tRNA and aaRS effectively both in aminoacylation and in editing. PMID- 12771212 TI - The role of intercalating residues in chromosomal high-mobility-group protein DNA binding, bending and specificity. AB - Ubiquitous high-mobility-group (HMGB) chromosomal proteins bind DNA in a non sequence- specific fashion to promote chromatin function and gene regulation. Minor groove DNA binding of the HMG domain induces substantial DNA bending toward the major groove, and several interfacial residues contribute by DNA intercalation. The role of the intercalating residues in DNA binding, bending and specificity was systematically examined for a series of mutant Drosophila HMGB (HMG-D) proteins. The primary intercalating residue of HMG-D, Met13, is required both for high-affinity DNA binding and normal DNA bending. Leu9 and Tyr12 directly interact with Met13 and are required for HMG domain stability in addition to linear DNA binding and bending, which is an important function for these residues. In contrast, DNA binding and bending is retained in truncations of intercalating residues Val32 and Thr33 to alanine, but DNA bending is decreased for the glycine substitutions. Furthermore, substitution of the intercalating residues with those predicted to be involved in the specificity of the HMG domain transcription factors results in increased DNA affinity and decreased DNA bending without increased specificity. These studies reveal the importance of residues that buttress intercalating residues and suggest that features of the HMG domain other than a few base-specific hydrogen bonds distinguish the sequence-specific and non-sequence-specific HMG domain functions. PMID- 12771214 TI - The Drosophila Corto protein interacts with Polycomb-group proteins and the GAGA factor. AB - In Drosophila, PcG complexes provide heritable transcriptional silencing of target genes. Among them, the ESC/E(Z) complex is thought to play a role in the initiation of silencing whereas other complexes such as the PRC1 complex are thought to maintain it. PcG complexes are thought to be recruited to DNA through interaction with DNA binding proteins such as the GAGA factor, but no direct interactions between the constituents of PcG complexes and the GAGA factor have been reported so far. The Drosophila corto gene interacts with E(z) as well as with genes encoding members of maintenance complexes, suggesting that it could play a role in the transition between the initiation and maintenance of PcG silencing. Moreover, corto also interacts genetically with Trl, which encodes the GAGA factor, suggesting that it may serve as a mediator in recruiting PcG complexes. Here, we show that Corto bears a chromo domain and we provide evidence for in vivo association of Corto with ESC and with PC in embryos. Moreover, we show by GST pull-down and two-hybrid experiments that Corto binds to E(Z), ESC, PH, SCM and GAGA and co-localizes with these proteins on a few sites on polytene chromosomes. These results reinforce the idea that Corto plays a role in PcG silencing, perhaps by confering target specificity. PMID- 12771215 TI - Optimisation of the 10-23 DNAzyme-substrate pairing interactions enhanced RNA cleavage activity at purine-cytosine target sites. AB - The 10-23 RNA cleaving DNAzyme has been shown to cleave any purine-pyrimidine (RY) junction under simulated physiological conditions. In this study, we systematically examine the DNAzymes relative activity against different RY combinations in order to determine the hierarchy of substrate core dinucleotide sequence susceptibility. The reactivity of each substrate dinucleotide compared in the same background sequence with the appropriately matched DNAzyme was found to follow the scheme AU = GU >> or = GC >> AC. The relatively poor activity of the DNAzyme against AC and GC containing substrates was found to be improved substantially by modifications to the binding domain which subtly weaken its interaction with the substrate core. The most effective modification resulting in rate enhancement of up to 200-fold, was achieved by substitution of deoxyguanine with deoxyinosine such that the base pair interaction with the RNA substrates core C is reduced from three hydrogen bonds to two. The increased cleavage activity generated by this modification could be important for application of the 10-23 DNAzyme particularly when the target site core is an AC dinucleotide. PMID- 12771216 TI - Experimental and computational analysis of transcriptional start sites in the cyanobacterium Prochlorococcus MED4. AB - In contrast to certain model eubacteria, little is known as to where transcription is initiated in the genomes of cyanobacteria, which are largely distinct from other prokaryotes. In this work, 25 transcription start sites (TSS) of 21 different genes of Prochlorococcus sp. MED4 were determined experimentally. The data suggest more than one TSS for the genes ftsZ, petH, psbD and ntcA. In contrast, the rbcL-rbcS operon encoding ribulose 1,5-bisphosphate carboxylase/oxygenase lacks a detectable promoter and is co-transcribed with the upstream located gene ccmK. The entire set of experimental data was used in a genome-wide scan for putative TSS in Prochlorococcus. A -10 element could be defined, whereas at the -35 position there was no element common to all investigated sequences. However, splitting the data set into sub-classes revealed different types of putative -35 boxes. Only one of them resembled the consensus sequence TTGACA recognized by the vegetative sigma factor (sigma70) of enterobacteria. Using a scoring matrix of the -10 element, more than 3000 TSS were predicted, about 40% of which were estimated to be functional. This is the first systematic study of transcription initiation sites in a cyanobacterium. PMID- 12771217 TI - ZBP-89 represses vimentin gene transcription by interacting with the transcriptional activator, Sp1. AB - Vimentin, a member of the intermediate filament protein family, is regulated both developmentally and tissue specifically. It is also a marker of the metastatic potential of many tumor cells. Pre viously, the human vimentin promoter has been shown to contain multiple elements for the binding of both positive- and negative acting regulatory factors. Transient transfection analysis of various vimentin 5' end promoter sequences and mutants thereof fused to a reporter gene further defined two regulatory elements, a positive element that binds Sp1 and a negative element that binds the protein ZBP-89. ZBP-89 has been shown to be either a repressor or an activator of gene expression, depending on the promoter. Here, we show that for vimentin, both ZBP-89 and ZBP-99 repress reporter gene expression in Schneider (S2) cells. Deletion constructs confirm that the glutamine-rich region of Sp1 is required to enhance vimentin transcription, whereas the N terminus of ZBP-89 is required to interact with Sp1 and repress gene expression. The overexpression of hTAF(II)130 can alleviate ZBP-89 repression in S2 cells, suggesting how ZBP-89 might serve to block gene expression. PMID- 12771218 TI - A new member of the MCM protein family encoded by the human MCM8 gene, located contrapodal to GCD10 at chromosome band 20p12.3-13. AB - The MCM8 protein from HeLa cells, a new member of the MCM family, co-isolates through several steps with MCM6 and MCM7, and MCM8 co-immunoprecipitates with MCM4, MCM6 and MCM7, proteins reportedly forming a helicase complex involved in initiation of DNA replication. MCM8 mRNA is expressed in placenta, lung and liver, but is also significantly expressed in adult heart, a tissue with a low percentage of proliferating cells. The MCM8 gene, consisting of 19 exons, is located contrapodal to a gene, consisting of 11 exons, encoding a homolog of the yeast GCD10 gene product. The region between these two transcription units, comprising as few as 62 bp, is TATA-less and highly GC-rich, containing multiple CpG units. MCM8 expression is altered in certain forms of neoplasia. In a case of choriocarcinoma MCM8 mRNA is aberrant, leading to expression of a protein lacking 16 amino acids. In several cases of colon adenocarcinoma MCM8 expression is greatly reduced relative to matched non-cancerous tissue. The potential helicase domain of MCM8 is different from those of other MCM proteins in that it is more homologous to canonical ATP-binding domains of other known helicases. Results suggest that MCM8 may interact with other MCM proteins to alter the function of the replicative MCM protein complex. PMID- 12771219 TI - Exploring the repertoire of RNA secondary motifs using graph theory; implications for RNA design. AB - Understanding the structural repertoire of RNA is crucial for RNA genomics research. Yet current methods for finding novel RNAs are limited to small or known RNA families. To expand known RNA structural motifs, we develop a two dimensional graphical representation approach for describing and estimating the size of RNA's secondary structural repertoire, including naturally occurring and other possible RNA motifs. We employ tree graphs to describe RNA tree motifs and more general (dual) graphs to describe both RNA tree and pseudoknot motifs. Our estimates of RNA's structural space are vastly smaller than the nucleotide sequence space, suggesting a new avenue for finding novel RNAs. Specifically our survey shows that known RNA trees and pseudoknots represent only a small subset of all possible motifs, implying that some of the 'missing' motifs may represent novel RNAs. To help pinpoint RNA-like motifs, we show that the motifs of existing functional RNAs are clustered in a narrow range of topological characteristics. We also illustrate the applications of our approach to the design of novel RNAs and automated comparison of RNA structures; we report several occurrences of RNA motifs within larger RNAs. Thus, our graph theory approach to RNA structures has implications for RNA genomics, structure analysis and design. PMID- 12771220 TI - Specific interactions of distamycin with G-quadruplex DNA. AB - Distamycin binds the minor groove of duplex DNA at AT-rich regions and has been a valuable probe of protein interactions with double-stranded DNA. We find that distamycin can also inhibit protein interactions with G-quadruplex (G4) DNA, a stable four-stranded structure in which the repeating unit is a G-quartet. Using NMR, we show that distamycin binds specifically to G4 DNA, stacking on the terminal G-quartets and contacting the flanking bases. These results demonstrate the utility of distamycin as a probe of G4 DNA-protein interactions and show that there are (at least) two distinct modes of protein-G4 DNA recognition which can be distinguished by sensitivity to distamycin. PMID- 12771221 TI - A novel engineered meganuclease induces homologous recombination in yeast and mammalian cells. AB - Homologous gene targeting is the ultimate tool for reverse genetics, but its use is often limited by low efficiency. In a number of recent studies, site- specific DNA double-strand breaks (DSBs) have been used to induce efficient gene targeting. Engineering highly specific, dedicated DNA endonucleases is the key to a wider usage of this technology. In this study, we present two novel, chimeric meganucleases, derived from homing endonucleases. The first one is able to induce recombination in yeast and mammalian cells, whereas the second cleaves a novel (chosen) DNA target site. These results are a first step toward the generation of custom endonucleases for the purpose of targeted genome engineering. PMID- 12771222 TI - Efficient clustering of large EST data sets on parallel computers. AB - Clustering expressed sequence tags (ESTs) is a powerful strategy for gene identification, gene expression studies and identifying important genetic variations such as single nucleotide polymorphisms. To enable fast clustering of large-scale EST data, we developed PaCE (for Parallel Clustering of ESTs), a software program for EST clustering on parallel computers. In this paper, we report on the design and development of PaCE and its evaluation using Arabidopsis ESTs. The novel features of our approach include: (i) design of memory efficient algorithms to reduce the memory required to linear in the size of the input, (ii) a combination of algorithmic techniques to reduce the computational work without sacrificing the quality of clustering, and (iii) use of parallel processing to reduce run-time and facilitate clustering of larger data sets. Using a combination of these techniques, we report the clustering of 168 200 Arabidopsis ESTs in 15 min on an IBM xSeries cluster with 30 dual-processor nodes. We also clustered 327 632 rat ESTs in 47 min and 420 694 Triticum aestivum ESTs in 3 h and 15 min. We demonstrate the quality of our software using benchmark Arabidopsis EST data, and by comparing it with CAP3, a software widely used for EST assembly. Our software allows clustering of much larger EST data sets than is possible with current software. Because of its speed, it also facilitates multiple runs with different parameters, providing biologists a tool to better analyze EST sequence data. Using PaCE, we clustered EST data from 23 plant species and the results are available at the PlantGDB website. PMID- 12771223 TI - A novel helper phage that improves phage display selection efficiency by preventing the amplification of phages without recombinant protein. AB - Phage display is a widely used technology for the isolation of peptides and proteins with specific binding properties from large libraries of these molecules. A drawback of the common phagemid/helper phage systems is the high infective background of phages that do not display the protein of interest, but are propagated due to non-specific binding to selection targets. This and the enhanced growth rates of bacteria harboring aberrant phagemids not expressing recombinant proteins leads to a serious decrease in selection efficiency. Here we describe a VCSM13-derived helper phage that circumvents this problem, because it lacks the genetic information for the infectivity domains of phage coat protein pIII. Rescue of a library with this novel CT helper phage yields phages that are only infectious when they contain a phagemid-encoded pIII-fusion protein, since phages without a displayed protein carry truncated pIII only and are lost upon re infection. Importantly, the CT helper phage can be produced in quantities similar to the VCSM13 helper phage. The superiority of CT over VCSM13 during selection was demonstrated by a higher percentage of positive clones isolated from an antibody library after two selection rounds on a complex cellular target. We conclude that the CT helper phage considerably improves the efficiency of selections using phagemid-based protein libraries. PMID- 12771224 TI - Use of a three-color cDNA microarray platform to measure and control support bound probe for improved data quality and reproducibility. AB - Construction methodologies for cDNA microarrays lack the ability to determine array integrity prior to hybridization, leaving the array itself a source of uncontrolled experimental variation. We solved this problem through development of a three-color cDNA array platform whereby printed probes are tagged with fluorescein and are compatible with Cy3 and Cy5 target labeling dyes when using confocal laser scanners possessing narrow bandwidths. Here we use this approach to: (i) develop a tracking system to monitor the printing of probe plates at predicted coordinates; (ii) define the quantity of immobilized probe necessary for quality hybridized array data to establish pre-hybridization array selection criteria; (iii) investigate factors that influence probe availability for hybridization; and (iv) explore the feasibility of hybridized data filtering using element fluorescein intensity. A direct and significant relationship (R2 = 0.73, P < 0.001) between pre-hybridization average fluorescein intensity and subsequent hybridized replicate consistency was observed, illustrating that data quality can be improved by selecting arrays that meet defined pre-hybridization criteria. Furthermore, we demonstrate that our three-color approach provides a means to filter spots possessing insufficient bound probe from hybridized data sets to further improve data quality. Collectively, this strategy will improve microarray data and increase its utility as a sensitive screening tool. PMID- 12771225 TI - Precise determination of mitochondrial DNA copy number in human skeletal and cardiac muscle by a PCR-based assay: lack of change of copy number with age. AB - Deletions in mitochondrial DNA (mtDNA) accumulate with age in humans without overt mitochondriopathies, but relatively limited attention has been devoted to the measurement of the total number of mtDNA molecules per cell during ageing. We have developed a precise assay that determines mtDNA levels relative to nuclear DNA using a PCR-based procedure. Quantification was performed by reference to a single recombinant plasmid standard containing a copy of each target DNA sequence (mitochondrial and nuclear). Copy number of mtDNA was determined by amplifying a short region of the cytochrome b gene (although other regions of mtDNA were demonstrably useful). Nuclear DNA content was determined by amplification of a segment of the single copy beta-globin gene. The copy number of mtDNA per diploid nuclear genome in myocardium was 6970 +/- 920, significantly higher than that in skeletal muscle, 3650 +/- 620 (P = 0.006). In both human skeletal muscle and myocardium, there was no significant change in mtDNA copy number with age (from neonates to subjects older than 80 years). This PCR-based assay not only enables accurate determination of mtDNA relative to nuclear DNA but also has the potential to quantify accurately any DNA sequence in relation to any other. PMID- 12771226 TI - A novel multiplex quantitative DNA array based PCR (MQDA-PCR) for quantification of transgenic maize in food and feed. AB - We have developed a novel multiplex quantitative DNA array based PCR method (MQDA PCR). The MQDA-PCR is general and may be used in all areas of biological science where simultaneous quantification of multiple gene targets is desired. We used quantification of transgenic maize in food and feed as a model system to show the applicability of the method. The method is based on a two-step PCR. In the first few cycles bipartite primers containing a universal 5' 'HEAD' region and a 3' region specific to each genetically modified (GM) construct are employed. The unused primers are then degraded with a single-strand DNA-specific exonuclease. The second step of the PCR is run containing only primers consisting of the universal HEAD region. The removal of the primers is essential to create a competitive, and thus quantitative PCR. Oligo nucleotides hybridising to internal segments of the PCR products are then sequence specifically labelled in a cyclic linear signal amplification reaction. This is done both to increase the sensitivity and the specificity of the assay. Hybridisation of the labelled oligonucleotides to their complementary sequences in a DNA array enables multiplex detection. Quantitative information was obtained in the range 0.1-2% for the different GM constructs tested. Seventeen different food and feed samples were screened using a twelve-plex system for simultaneous detection of seven different GM maize events (Bt176, Bt11, Mon810, T25, GA21, CBH351 and DBT418). Ten samples were GM positive containing mainly mixtures of Mon810, Bt11 and Bt176 DNA. One sample contained appreciable amounts of GA21. An eight-plex MQDA-PCR system for detection of Mon810, Bt11 and Bt176 was evaluated by comparison with simplex 5' nuclease PCRs. There were no significant differences in the quantifications using the two approaches. The samples could, by both methods, be quantified as containing >2%, between 1 and 2%, between 0.1 and 1%, or <0.1% in 43 out of 47 determinations. The described method is modular, and thus suited for future needs in GM detection. PMID- 12771227 TI - MALDI mass spectrometry analysis of single nucleotide polymorphisms by photocleavage and charge-tagging. AB - High-throughput procedures are an important requirement for future large-scale genetic studies such as genotyping of single nucleotide polymorphisms (SNPs). Matrix-assisted laser desorption/ ionisation mass spectrometry (MALDI-MS) has revolutionised the analysis of biomolecules and, in particular, provides a very attractive solution for the rapid typing of DNA. The analysis of DNA by MALDI can be significantly facilitated by a procedure termed 'charge-tagging'. We show here a novel approach for the generation of charge-tagged DNA using a photocleavable linker and its implementation in a molecular biological procedure for SNP genotyping consisting of PCR, primer extension, photocleavage and a chemical reaction prior to MALDI target preparation and analysis. The reaction sequence is amenable to liquid handling automation and requires no stringent purification procedures. We demonstrate this new method on SNPs in two genes involved in complex traits. PMID- 12771228 TI - In vitro analysis of nuclear mRNA export using molecular beacons for target detection. AB - A detailed molecular characterization of nuclear mRNA export will require an in vitro system, allowing a biochemical reconstitution of transport. To this end, an mRNA export assay has been developed using digitonin-permeabilized HeLa cells and 2'-O-methyl oligoribonucleotide molecular beacons for target detection. These probes allow the homogeneous detection of poly(A)+ RNA at subnanomolar concentrations in the presence of cytosol, without the need for RNA purification and time-consuming methods like northern blotting or RT-PCR. Nuclear export of endogenous mRNA in permeabilized cells occurs in a time- and temperature dependent manner and can be inhibited by wheat germ agglutinin, indicative of specific transport through nuclear pore complexes. Nuclear export in vitro is insensitive to the depletion of ATP and does not depend on the addition of cytosolic factors, suggesting that shuttling proteins are not required for efficient transport. This is the first demonstration of molecular beacons as powerful tools for the analysis of nucleocytoplasmic RNA transport. PMID- 12771229 TI - An automated microplate-based method for monitoring DNA strand breaks in plasmids and bacterial artificial chromosomes. AB - A method is described for high-throughput monitoring of DNA backbone integrity in plasmids and artificial chromosomes in solution. The method is based on the denaturation properties of double-stranded DNA in alkaline conditions and uses PicoGreen fluorochrome to monitor denaturation. In the present method, fluorescence enhancement of PicoGreen at pH 12.4 is normalised by its value at pH 8 to give a ratio that is proportional to the average backbone integrity of the DNA molecules in the sample. A good regression fit (r2 > 0.98) was obtained when results derived from the present method and those derived from agarose gel electrophoresis were compared. Spiking experiments indicated that the method is sensitive enough to detect a proportion of 6% (v/v) molecules with an average of less than two breaks per molecule. Under manual operation, validation parameters such as inter-assay and intra-assay variation gave values of <5% coefficient of variation. Automation of the method showed equivalence to the manual procedure with high reproducibility and low variability within wells. The method described requires as little as 0.5 ng of DNA per well and a 96-well microplate can be analysed in 12 min providing an attractive option for analysis of high molecular weight vectors. A preparation of a 116 kb bacterial artificial chromosome was subjected to chemical and shear degradation and DNA integrity was tested using the method. Good correlation was obtained between time of chemical degradation and shear rate with fluorescence response. Results obtained from pulsed- field electrophoresis of sheared samples were in agreement with those obtained using the microplate-based method. PMID- 12771230 TI - Genotyping single nucleotide polymorphisms directly from genomic DNA by invasive cleavage reaction on microspheres. AB - Here we report proof-of-principle for a microsphere-based genotyping assay that detects single nucleotide polymorphisms (SNPs) directly from human genomic DNA samples. This assay is based on a structure-specific cleavage reaction that achieves single base discrimination with a 5'-nuclease which recognizes a tripartite substrate formed upon hybridization of target DNA with probe and upstream oligonucleotides. The assay is simple with two easy steps: a cleavage reaction, which generates fluorescent signal on microsphere surfaces, followed by flow cytometry analysis of the microspheres. Genomic DNA samples were genotyped for the SNP in the Apolipoprotein E gene at amino acid position 158. The assay successfully scored wild type, heterozygous and homozygous mutants. To our knowledge, this is the first report of a solid-support assay for detection of SNPs directly from genomic DNA without PCR amplification of the target. PMID- 12771231 TI - Melanoma cell adhesion molecule (MCAM/CD146) is expressed on human luteinizing granulosa cells: enhancement of its expression by hCG, interleukin-1 and tumour necrosis factor-alpha. AB - Melanoma cell adhesion molecule (MCAM) was originally reported to be involved in the invasion and progression of melanoma. It was also shown to be responsible for the attachment of cells to endothelial cells. In this study, we demonstrated by immunohistochemistry that immunoreactive MCAM was not expressed on granulosa cells in the pre-ovulatory follicle, but it was clearly detected in large luteal cells in corpora lutea from the mid-luteal phase of the menstrual cycle. Northern blotting analysis confirmed the expression of MCAM mRNA in corpus luteum. MCAM was weakly detected by immunocytochemical staining in human luteinizing granulosa cells isolated from patients undergoing IVF treatment. Its expression was found to be increased during time in culture of these cells. Flow cytometry and Northern blot analysis revealed that MCAM expression on luteinizing granulosa cells was enhanced when the cells were cultured for 5 days in the presence of hCG (1 IU/ml) or cytokines such as interleukin-1alpha (10 ng/ml) and tumour necrosis factor-alpha (10 ng/ml). No significant difference of MCAM expression was observed between the cultures under normoxic (20% oxygen) and hypoxic (1% oxygen) conditions. These results indicate that luteinizing granulosa cells express MCAM and that MCAM expression is regulated by LH/hCG and cytokines during luteinization. Since MCAM has been reported to mediate cellular interaction with endothelial cells, this molecule may play a role in neovascularization during corpus luteum formation in the human ovary. PMID- 12771232 TI - Characterization, expression pattern and chromosomal localization of the spermatogenesis associated 6 gene (Spata6). AB - We report the cloning and characterization of the spermatogenesis associated 6 gene (Spata6) encoding a predicted protein of 488 amino acids. It exhibits similarity with the motor domain of kinesin related proteins and with the Caenorhabditis elegans neural calcium sensor protein (NCS-2). The gene encodes three mRNAs of approximately 2.6, approximately 1.8 and approximately 1.2 kb. The expression of the 2.6 kb mRNA is detected at low levels in testis, ovary, thymus and placenta, while the 1.8 and 1.2 kb transcripts are exclusively expressed in testis. The 1.8 and 1.2 kb transcripts are specifically expressed in haploid germ cells. Data from in situ hybridization experiments suggested that mRNA expression of Spata6 in spermatids is higher than in spermatocytes and spermatogonia. RT-PCR analysis and whole mount in situ hybridization demonstrate that the Spata6 transcript is expressed during embryonic development and is localized in neural tube, somites and limb buds of mouse embryo. The Spata6 gene consists of 15 exons ranging in size between 40 and 596 bp. The 2.6 and 1.8 kb transcripts have different 5' untranslated sequences but have the same translational initiation site and therefore may encode the same protein with a predicted molecular weight of 49.7 kDa. The 1.2 kb transcript is derived from a proximal promoter between exons 7 and 8, and contains a translation initiation codon AUG, which is in frame with initiator AUG codon of the 2.6 and 1.8 kb transcripts. Therefore, the 1.2 kb transcript may code for a truncated protein of 32 kDa. Western blot analysis with the antiserum raised against a synthetic peptide from the C-terminal of the deduced Spata6 protein detects only a single protein of 53 kDa in all tissues studied. The Spata6 gene was localized to chromosome 5, region q34-35 in the rat and to chromosome 1, region p32-35 in the human. In an effort to determine the function of Spata6, we inactivated the mouse gene in embryonic stem cells through homologous recombination. Although the heterozygous mutant cells were able to generate low coat colour chimeric mice, all chimeras did not transmit the targeted allele to their progeny suggesting that a high contribution of Spata6(+/ ) cells lead to the lethality of the chimeric embryos. PMID- 12771233 TI - Decreased protamine-1 transcript levels in testes from infertile men. AB - Infertile men exhibit an aberrant protamine-1 (Prm1) to protamine-2 (Prm2) ratio at both the mRNA and protein level. We therefore investigated whether male infertility could be related to the amount of Prm1 and Prm2 mRNA by applying real time quantitative PCR following RNA extraction from routinely Bouin-fixed and paraffin-embedded testicular biopsies. Samples (n = 51) were normalized to the same amount and similar size of tissue sections. The threshold cycle (C(T)) representing a measure of the initial number of mRNA copies was significantly (P < 0.001) higher for Prm1, but not Prm2, and thus the amount of Prm1 mRNA was lower in men with at least qualitatively normal spermatogenesis (Prm1: 29.88 +/- 2.99; Prm2: 34.28 +/- 2.26) and impaired spermatogenesis (Prm1: 31.89 +/- 2.54; Prm2: 35.59 +/- 2.09) compared with men with obstructive azoospermia and quantitatively normal spermatogenesis (Prm1: 29.04 +/- 1.02; Prm2: 34.91 +/- 1.40). In addition, the Prm1 - Prm2 C(T) difference (deltaC(T)) was significantly (P < 0.001) decreased in these two groups. A negative correlation (r = -0.504; P < 0.001) was demonstrated between the score for efficiency of spermatogenesis and the C(T) for Prm1. These data suggest that the decreasing amount of Prm1 and, as a consequence, the aberrant Prm1:Prm2 mRNA ratio plays an important role for male infertility and may serve as a possible predictive factor for the outcome of ICSI. PMID- 12771234 TI - Epitope analysis for human sperm-immobilizing monoclonal antibodies, MAb H6-3C4, 1G12 and campath-1. AB - Human monoclonal antibody, MAb H6-3C4, possesses strong sperm immobilizing activity. MAb H6-3C4 has been suggested by several research groups to react with a carbohydrate moiety of male reproductive tract CD52 (mrtCD52). In the present study, we analysed the epitope on mrtCD52 for MAb H6-3C4 and found that it was polymorphic in Western blot analysis and disappeared after enzymatic removal of the N-linked carbohydrate moiety. Two other monoclonal antibodies (1G12, campath 1) with sperm-immobilizing activity recognized mrtCD52 in a polymorphic manner similar to MAb H6-3C4. Further analysis showed that 1G12 recognized a structure formed by the peptide and/or a glycosylphosphatidylinositol (GPI) anchor portion as does campath-1. Results of a lectin binding assay suggested the presence of O linked carbohydrates on mrtCD52. Our results also indicated that the peptide portion of CD52 could serve as an epitope for sperm-immobilizing antibodies. It was concluded that the epitope of MAb H6-3C4 is similar to, but distinct from, those of 1G12 and campath-1, and that mrtCD52 contains different antigenic epitopes. PMID- 12771235 TI - A frequent allele codes for a truncated variant of semenogelin I, the major protein component of human semen coagulum. AB - Human semen coagulum predominantly consists of high molecular mass complexes of the seminal vesicle secreted semenogelin I (SgI) and semenogelin II (SgII). Here we describe a previously unknown variant of the SgI gene that is present at an allele frequency of approximately 3% in the Swedish population. It gives rise to a protein with a molecular mass of 43 kDa, SgI(43), which compared with the 50 kDa variant, SgI(50), is lacking a tandem repeat of 60 amino acid residues that was probably deleted by homologous recombination. In spite of the size difference, SgI(43) has many properties in common with SgI(50), such as a very high iso-electric point and susceptibility to proteolytic degradation by prostate specific antigen. Heterozygous carriers of the SgI(43) allele neither show impaired fertility nor do they significantly differ from individuals homozygous for SgI(50) with respect to sperm parameters such as semen volume, sperm count and fraction of motile spermatozoa. PMID- 12771236 TI - Gestational profile of matrix metalloproteinases in rat uterine artery. AB - Mechanisms underlying structural reorganization of the uterine artery in pregnancy remain largely unknown. Matrix metalloproteinases (MMPs) which are involved in degradation of vascular wall matrix are likely to play a key role. In this investigation of rat uterine artery, key MMPs and the specific tissue inhibitors of MMPs (TIMPs) together with three housekeeping genes were studied before, during and after pregnancy, using real time PCR. Data were analysed by partial least squares analysis as well as by conventional univariate methods. Each gene studied [MMP-2, MMP-3, MMP-7, MMP-9, MMP-12, MMP-13, membrane-type 1 (MT1)-MMP, TIMP-1, TIMP-2, GAPDH, cyclophilin and beta-actin] increased in late pregnancy (day 21). MMP-2, MT1MMP, MMP-3 and TIMP-1 transcripts were also elevated at day 7. TIMP-1 and MMP-3 mRNA expression returned to virgin control values in the post-partum, whereas others remained elevated or increased further (MMP-9, MMP-13). Gelatin zymography showed maximum elevation of MMP-2 at day 21. A novel 43-45 kDa gelatinolytic doublet was observed which increased in density with gestation and may represent an active MMP-2 fragment. Together, these data strongly suggest that MMPs and TIMPs are likely to play an important role in remodelling uterine arteries in rat pregnancy and may represent means by which vasodilatation is maintained in later pregnancy. Continued elevated levels of some MMPs post-partum may contribute to vessel regression and return to a non pregnant physiological state. PMID- 12771237 TI - Cyclic AMP enhances the expression of an extravillous trophoblast marker, melanoma cell adhesion molecule, in choriocarcinoma cell JEG3 and human chorionic villous explant cultures. AB - Human trophoblasts consist of two main cell lineages, villous trophoblasts (VT) and extravillous trophoblasts (EVT). To identify the molecules which are involved in EVT differentiation, we have raised a monoclonal antibody (mAb) designated CHL1, by immunizing a mouse against human chorion laeve which is composed of EVT. By immunohistochemical analysis, the CHL1 antigen was found to be expressed on the majority of EVT but not on VT in addition to its expression on endothelial and myometrial cells. A subsequent cDNA panning method revealed that the CHL1 antigen was identical to melanoma cell adhesion molecule (MCAM, Mel-CAM, S-endo 1 or MUC18/CD146), which has been previously reported as one of the EVT markers. MCAM expression on JEG3 cells, a human choriocarcinoma-derived cell line, was significantly enhanced when they were co-cultured with isolated human decidual tissue. Various cytokines and growth factors that were reportedly present in decidual tissue failed to increase MCAM expression in JEG3 cells, but decidua induced MCAM expression in JEG3 cells was attenuated by the addition of protein kinase A inhibitor H89. In addition, cAMP, which is known to stimulate differentiation of VT, enhanced MCAM expression in JEG3 cells. Its promoting effect on MCAM expression was also observed in human chorionic villous explant cultures. These findings suggest that a cAMP-dependent intracytoplasmic signalling pathway is involved in the differentiation mechanism of human EVT. PMID- 12771238 TI - Differential activity of the gelatinases (matrix metalloproteinases 2 and 9) in the fetal membranes and decidua, associated with labour. AB - Degradation of the extracellular matrix in fetal membranes has been implicated in the rupture of fetal membranes, the process of parturition and placental detachment from the decidua after parturition. In this study we assessed labour associated changes in gelatinase activity in cultured human amnion, chorion and decidua, as well as in amniotic fluid. We found that in media conditioned by decidua, following the establishment of uterine contractions, matrix metalloproteinase-2 (MMP-2) activity is increased while the protein tissue inhibitors of matrix metalloproteinase-1 (TIMP-1) level is decreased. The formation of a 130 kDa gelatinase band was also significantly increased after contractions began. In media conditioned by chorion, the initiation of uterine contractions did not change MMP activity or TIMP-1 levels. However, an increase in MMP-9 activity and a decrease in TIMP-1 protein levels were observed following the establishment of uterine contractions in media conditioned by amnion. We suggest that this differential spatial regulation provides a form for modulatory hieratical activity of the MMPs in the onset of labour allowing rupture of the membranes while avoiding premature placental separation. PMID- 12771239 TI - Neurologic manifestations of celiac disease: proven, or just a gut feeling? PMID- 12771240 TI - Idebenone for treatment of Friedreich's ataxia? PMID- 12771241 TI - Dementia with Lewy bodies: improving diagnostic criteria. PMID- 12771242 TI - Is language laterality established by 5 years of age? PMID- 12771244 TI - Stroke following Bothrops spp. snakebite. AB - OBJECTIVE: To determine the prevalence and subtypes of cerebrovascular complications of Bothrops spp. snakebites. METHODS: The authors studied 309 consecutive patients bitten by Bothrops spp. who attended a large general hospital. Special emphasis was placed on the time elapsed between the bite and admission to the hospital, clinical manifestations, laboratory findings, severity of envenomation, CT findings, and outcome. RESULTS: Eight of 309 patients (2.6%) developed a cerebrovascular event. Six presented >8 hours after being bitten, and all had clinical and laboratory evidence of severe envenoming. CT revealed intracranial bleeding in seven patients and multiple brain infarcts in one. Hemorrhages were located in the subcortical white matter of the cerebral hemispheres in five patients, in the cerebellum in one, and in the subarachnoid space in one. All patients with intracranial hemorrhage had a bleeding disorder, and the patient with cerebral infarcts may have had angiitis. Five of the eight patients died despite therapy, and the remaining three had irreversible sequelae. CONCLUSIONS: The prevalence of cerebrovascular complications related to Bothrops spp. bites is 2.6%. Intracranial hemorrhages are more frequent than cerebral infarcts. The prognosis of these patients is poor. PMID- 12771245 TI - Celiac neuropathy. AB - BACKGROUND: Celiac disease (CD) is a chronic inflammatory enteropathy resulting from sensitivity to ingested gluten. Neurologic complications are estimated to occur in 10% of affected patients, with ataxia and peripheral neuropathy being the most common problems. The incidence and clinical presentation of patients with CD-associated peripheral neuropathy have not previously been investigated. OBJECTIVE: To determine the incidence of CD in patients with neuropathy and to characterize the clinical presentation. METHODS: The records of 20 patients with neuropathy and biopsy-confirmed CD were reviewed. RESULTS: Six of the 20 patients had neuropathic symptoms alone without gastrointestinal involvement, and neuropathic symptoms preceded other CD symptoms in another 3 patients. All patients had burning, tingling, and numbness in their hands and feet, with distal sensory loss, and nine had diffuse paresthesias involving the face, trunk, or lumbosacral region. Only two had weakness. Results of electrophysiologic studies were normal or mildly abnormal in 18 (90%) of the patients. Sural nerve biopsies, obtained from three patients, revealed mild to severe axonopathy. Using the agglutination assay, 13 (65%) of the patients were positive for ganglioside antibodies. Excluding patients who were referred with the diagnosis of celiac neuropathy, CD was seen in approximately 2.5% of all neuropathy patients and in 8% of patients with neuropathy and normal electrophysiologic studies seen at our center. CONCLUSION: CD is commonly associated with sensory neuropathy and should be considered even in the absence of gastrointestinal symptoms. PMID- 12771246 TI - Influence of Alzheimer pathology on clinical diagnostic accuracy in dementia with Lewy bodies. AB - OBJECTIVE: To determine whether AD neurofibrillary pathology influences clinical diagnostic accuracy in dementia with Lewy bodies (DLB). BACKGROUND: Pathologic diagnosis of DLB mandates Lewy bodies but also allows for AD pathology in the form of plaques and tangles. Because clinical diagnostic accuracy of DLB remains low, the authors questioned whether the severity of AD pathology in the form of tangles might affect the clinician's ability to correctly diagnose DLB in life. DESIGN/METHODS: Ninety-eight subjects with autopsy-proven DLB who had been evaluated annually at the University of California San Diego AD Research Center were identified. The clinical diagnosis used was the last diagnosis before death. Pathologic diagnosis of DLB was made according to Consensus guidelines, and Braak staging was used to assess the degree of neurofibrillary AD pathology. The clinical characteristics of subjects with DLB with low vs high Braak stages were compared and the clinical diagnostic accuracy for subjects stratified according to Braak stage was determined. RESULTS: Only 27% of the subjects with DLB demonstrated both visual hallucinations and spontaneous extrapyramidal signs (EPS). The low Braak stage (0 to 2, n = 24) subjects had a higher frequency of visual hallucinations (65%) than did subjects with DLB with higher (3 to 6, n = 66) Braak stages (33%, p = 0.008), and showed a slightly greater but not significant degree of EPS. Although clinical diagnostic accuracy for DLB was relatively low (49%), it was higher for subjects with low (75%) compared to high (39%) Braak stages (p = 0.0039). CONCLUSIONS: The degree of concomitant AD tangle pathology has an important influence on the clinical characteristics and, therefore, the clinical diagnostic accuracy of DLB. PMID- 12771247 TI - Regular use of nonsteroidal anti-inflammatory drugs and cognitive function in aging women. AB - OBJECTIVE: To examine the relationship of nonsteroidal anti-inflammatory drug (NSAID) use and cognitive decline in young-old women. METHODS: The authors prospectively studied 16,128 Nurses' Health Study participants, aged 70 to 81 years at baseline, who provided information on NSAID use and potential confounders in biennial questionnaires from 1976 through 1998. From 1995 through 2001, we administered, by telephone, six tests of cognitive function, including the Telephone Interview of Cognitive Status (TICS). Second interviews were begun 2 years later and completed on 13,255 women to date. The authors used multiple logistic regression to estimate relative risks (RR) of low baseline scores (defined as the bottom 10%) and substantial decline (worst 10%). RESULTS: Compared to never users, the RR was 0.75 (95% CI 0.59, 0.96) for a low baseline TICS score with current aspirin use of 15+ years duration, and 0.79 (95% CI 0.62, 1.02) for current use of NSAID (primarily ibuprofen) lasting 8+ years. Results for aspirin users were weaker on other tests, but long-term ibuprofen users had a RR of 0.75 (95% CI 0.56, 1.00) for a low baseline global score (combination of all six tests). The RR for substantial global cognitive decline was 0.93 (95% CI 0.68, 1.26) with long-term aspirin use, and 0.77 (95% CI 0.57, 1.05) with long term ibuprofen use. CONCLUSIONS: In these young-old women, current, long-term NSAID users, especially of nonaspirin agents, showed reduced odds of low cognitive function and possibly lower rates of substantial cognitive decline over 2 years. Continued follow-up will help determine if associations differ at older ages. PMID- 12771248 TI - Auditory comprehension of language in young children: neural networks identified with fMRI. AB - OBJECTIVE: The organization of neuronal systems that process language in young children is poorly understood. The authors used fMRI to identify brain regions underlying auditory comprehension in healthy young children. METHODS: Fifteen right-handed children (mean age 6.8 years) underwent fMRI at 1.5-T using blood oxygen level dependent echoplanar imaging. They listened to stories with a reverse speech control condition. Group data were analyzed with statistical parametric mapping. Individual subject data were analyzed with a region of interest approach based on t-maps. An asymmetry index (AI = [(L-R)/(L+R)]) was calculated for each region. RESULTS: Group analysis showed significant activation in the left middle temporal gyrus (Brodmann area [BA] 21) and left superior temporal gyrus (BA 22) along the superior temporal sulcus extending back to the angular gyrus (BA 39). Individual maps showed lateralized activation in temporal regions (AI > 0.49 +/- 0.39). There was minimal activation in the frontal lobe. There were no significant correlations between age and regional AI. CONCLUSION: Networks for auditory language processing are regionally localized and lateralized by age 5. These data may provide a means to interpret language fMRI studies performed in preparation for brain surgery, and may be employed to investigate the effect of chronic disease states, such as epilepsy, on language organization during critical periods for plasticity. PMID- 12771249 TI - Parkin-proven disease: common founders but divergent phenotypes. AB - OBJECTIVE: To compare and contrast clinical and genetic findings in six probands with parkinsonism with a parkin exon 3 438- to 477-bp deletion (Ex3Delta40) to search for evidence of a common founder. METHOD: Clinical review, parkin gene sequencing, dosage studies, and high-resolution genotype/haplotype analysis were performed. RESULTS: All subjects had two or more signs consistent with a diagnosis of possible or probable PD with age at onset younger than 45 years (mean +/- SD 29.3 +/- 10.2 years, range 16 to 42 years). Affected individuals were either homozygotes, compound heterozygotes, or Ex3Delta40 carriers with one normal parkin allele. Haplotype analysis revealed both Ex3Delta40 and Ex7 924 C- >T (R275W) mutations originated from common founders, the former most probably of Irish descent. Although three cases had Ex7 924 C-->T (R275W) and Ex3Delta40 mutations, their clinical presentation and mode of inheritance were variable. CONCLUSION: Parkin mutations on common parkin haplotypes provide testable hypotheses of parkin function in genetically defined parkinsonism. PMID- 12771250 TI - Head trauma preceding PD: a case-control study. AB - OBJECTIVE: To investigate the association of PD with preceding head trauma using a case-control study design. METHODS: The medical records-linkage system of the Rochester Epidemiology Project was used to identify 196 subjects who developed PD in Olmsted County, MN, from 1976 through 1995. Each incident case was matched by age (+/-1 year) and sex to a general population control. The complete medical records of cases and controls in the system were reviewed to detect preceding episodes of head trauma. RESULTS: The frequency of head trauma overall was significantly higher in cases than in controls (odds ratio [OR] = 4.3; 95% CI = 1.2 to 15.2). Compared with subjects who never experienced a trauma, subjects who experienced a mild head trauma with only amnesia had no increased risk; however, subjects who experienced a mild head trauma with loss of consciousness or a more severe trauma had an OR of 11.0 (95% CI = 1.4 to 85.2). Although not significant, head trauma resulting in hospitalization was more frequent in cases than in control subjects (OR = 8.0; 95% CI = 1.0 to 64.0). Whereas the OR was higher for men than women and for patients with later onset of PD than for patients with earlier onset, these differences were not significant. CONCLUSIONS: These results suggest an association between head trauma and the later development of PD that varies with severity. Although the OR is high (4.3), the population attributable risk is only 5% because head trauma is a relatively rare event. PMID- 12771251 TI - Evidence for more widespread cerebral pathology in early HD: an MRI-based morphometric analysis. AB - BACKGROUND: Most clinical symptoms of Huntington disease (HD) have been attributed to striatal degeneration, but extrastriatal degeneration may play an important role in the clinical symptoms because postmortem studies demonstrate that almost all brain structures atrophy. OBJECTIVE: To fully characterize the morphometric changes that occur in vivo in HD. METHODS: High-resolution 1.5 mm T1 weighted coronal scans were acquired from 18 individuals in early to mid-stages of HD and 18 healthy age-matched controls. Cortical and subcortical gray and white matter were segmented using a semiautomated intensity contour-mapping algorithm. General linear models for correlated data of the volumes of brain regions were used to compare groups, controlling for age, education, handedness, sex, and total brain volumes. RESULTS: Subjects with HD had significant volume reductions in almost all brain structures, including total cerebrum, total white matter, cerebral cortex, caudate, putamen, globus pallidus, amygdala, hippocampus, brainstem, and cerebellum. CONCLUSIONS: Widespread degeneration occurs in early to mid-stages of HD, may explain some of the clinical heterogeneity, and may impact future clinical trials. PMID- 12771252 TI - Novel prion protein insert mutation associated with prolonged neurodegenerative illness. AB - BACKGROUND: Mutations in the prion protein gene (PRNP) are found in approximately 13 to 15% of persons classified as dying from a transmissible spongiform encephalopathy. Point and octapeptide repeat insert and deletion mutations are described in the open reading frame (ORF) of PRNP. The authors present a clinicopathologic study of a patient with a family history of a lengthy and progressive neurodegenerative disorder associated with a novel large octapeptide repeat insert mutation. METHODS: Neuropathologic examination, including immunohistochemistry for the prion protein, was undertaken. The ORF of PRNP was amplified by PCR, cloned, and sequenced. Homogenate of cerebral tissue underwent Western blot analysis for the prion protein before and after proteinase K treatment. RESULTS: The proband died after a 16-year illness commencing at age 29 years. Confident premortem clinical diagnosis was not achieved despite a brain biopsy. Autopsy examination of the brain confirmed a spongiform encephalopathy. Prion protein immunohistochemistry revealed occasional granular deposits in the cerebellar granular layer. The proband was found to harbor a novel PRNP 168 base pair (bp) insert mutation. CONCLUSION: The authors have identified a novel 168 bp octapeptide repeat insert mutation. Prion protein immunohistochemistry differs from previous cases harboring seven octapeptide repeat and other long insert mutations. Optimization of PRNP analysis, especially PCR conditions, is essential to avoid overlooking this type of mutation and delay the correct molecular genetic diagnosis. PMID- 12771253 TI - Oculoleptomeningeal amyloidosis in a large kindred with a new transthyretin variant Tyr69His. AB - OBJECTIVE: To describe the clinical, radiologic, and pathologic findings of a kindred with oculoleptomeningeal amyloidosis and a newly associated transthyretin mutation. BACKGROUND: Transthyretin (TTR) amyloidosis can present in the form of oculoleptomeningeal amyloidosis. Clinical features include dementia, seizures, stroke-like episodes, subarachnoid hemorrhage, ataxia, myelopathy, deafness, radiculopathy, and ocular amyloidosis. Eight TTR mutations associated with oculoleptomeningeal amyloidosis have been described. METHODS: Fourteen individuals from a kindred with oculoleptomeningeal amyloidosis were examined clinically and radiologically. Analysis of the TTR gene was performed. Neuropathologic examination was obtained on the index patient. RESULTS: Affected individuals had vitreous amyloid, radiculopathy, seizures, stroke-like episodes, encephalopathy, and dementia. Severely affected individuals died by the end of the fifth decade. Leptomeningeal enhancement on contrast MRI and elevated CSF protein were the defining features on investigations. Sequencing of exon 3 in the TTR gene found a base pair substitution at codon 69. This resulted in heterozygosity for normal tyrosine and variant histidine (ATTR Tyr69His) in affected family members. Domino liver transplantation was attempted as treatment for one family member. CONCLUSIONS: The ATTR Tyr69His mutation is associated with oculoleptomeningeal amyloidosis. Expression of the genotype is variable. This has implications for treatment of affected individuals and counseling of family members. Efficacy of liver transplantation in patients with oculoleptomeningeal amyloidosis remains unknown. The authors advocate the investigation of liver transplantation in patients with severe symptoms due to oculoleptomeningeal amyloidosis. PMID- 12771254 TI - Dose-response trial of pregabalin adjunctive therapy in patients with partial seizures. AB - BACKGROUND: Pregabalin is an alpha(2)-delta ligand that has anxiolytic, analgesic, and anticonvulsant properties. OBJECTIVE: To establish the efficacy, safety, and tolerability of pregabalin administered twice-daily (BID) without dose titration as adjunctive treatment in patients with partial seizures and to confirm the dose-response relationship. METHODS: This 76-center, double-blind, randomized, placebo-controlled, parallel-group study consisted of an 8-week baseline and a 12-week double-blind phase. Patients with refractory partial seizures on one to three antiepileptic drugs were randomly assigned to one of five treatment groups (placebo or 50, 150, 300, and 600 mg/d pregabalin, all administered BID). Efficacy was assessed using seizure frequency reduction and responder rate (> or =50% seizure reduction from baseline). Pharmacokinetic parameters were estimated. RESULTS: A total of 453 patients were included in the intent-to-treat analysis. The median baseline seizure rate was 10 per month. Seizure frequency reductions from baseline were 7% (placebo; n = 100), 12% (50 mg/d; n = 88), 34% (150 mg/d; n = 86), 44% (300 mg/d; n = 90), and 54% (600 mg/d; n = 89). Responder rates (> or =50% seizure reduction) were 14% (placebo), 15% (50 mg/d), 31% (150 mg/d), 40% (300 mg/d), and 51% (600 mg/d). Discontinuation rates due to adverse events were 5% (placebo), 7% (50 mg/d), 1% (150 mg/d), 14% (300 mg/d), and 24% (600 mg/d). The 150-, 300-, and 600-mg/d pregabalin groups were associated with greater reductions in seizures (p < or = 0.0001) and greater responder rates compared with the placebo group (p < or = 0.006). There was a favorable dose-response trend for both seizure reductions (p < or = 0.0001) and responder rate (p < or = 0.001). CONCLUSION: Adjunctive therapy with pregabalin 150, 300, and 600 mg/d, given in twice-daily doses without titration, is significantly effective and well tolerated in the treatment of patients with partial seizures as demonstrated in patients with refractory partial seizures. PMID- 12771255 TI - Magnetization transfer imaging in focal epilepsy. AB - OBJECTIVES: To test the hypothesis that magnetization transfer imaging (MTI), analyzed on a voxel-by-voxel basis, would identify areas of abnormal magnetization transfer ratio (MTR) in patients with focal epilepsy. METHODS: The authors used MTI maps and statistical parametric mapping (SPM) to objectively compare the cerebral structures of 15 patients with malformations of cortical development (MCD), 10 with partial seizures and acquired lesions, and 42 with partial seizures and normal conventional MRI with those of 30 control subjects. RESULTS: Significant reductions of MTR were identified in all 10 patients with acquired nonprogressive cerebral lesions and partial seizures. In all, the areas of decreased MTR concurred with abnormalities identified on visual inspection of conventional MRI. In 13 of the 15 patients with MCD, SPM detected regions of significantly reduced MTR, all of which corresponded to abnormalities identified on visual inspection of conventional MRI. In addition, in both groups, there were areas that were normal on conventional imaging that demonstrated abnormal MTR. There was a significant reduction of MTR in 15 of the 42 patients with cryptogenic focal epilepsy. In all of these, the areas of reduced MTR concurred with epileptiform EEG abnormality and clinical seizure semiology. CONCLUSIONS: Magnetization transfer imaging analyzed using statistical parametric mapping was sensitive in identifying malformations of cortical development and acquired cerebral lesions. Abnormalities of magnetization transfer ratio in individual MRI negative patients suggest that minor structural disorganization exists in occult epileptogenic cerebral lesions. PMID- 12771256 TI - N-acetyl-aspartate, total creatine, and myo-inositol in the epileptogenic human hippocampus. AB - BACKGROUND: Mesial temporal lobe epilepsy (mTLE) is characterized by hippocampal atrophy, decreased N-acetyl-aspartate, and a low N-acetyl-aspartate/total creatine ratio, often attributed to neuron loss and gliosis. Qualitative studies reported that N-acetyl-aspartate content was significantly lower in hippocampal sclerosis. OBJECTIVE: It was proposed to measure the effects of neuron loss and gliosis on the hippocampal content of N-acetyl-aspartate, total creatine, and myo inositol in mTLE. METHODS: Twenty hippocampal specimens were obtained during temporal lobectomy and frozen quickly. Perchloric acid extracts of the small metabolites were prepared and analyzed by proton MRS at 11.75 T. Adjacent samples were used for cell counts. RESULTS: There were no significant associations between hippocampal neuron loss and the cellular content of N-acetyl-aspartate, total creatine, or myo-inositol, despite more than a threefold difference in neuron loss and a twofold increase in glial density. Metabolite concentrations varied two- to fourfold. Variation in the cellular content of total creatine accounted for more than three-quarters of the rank-order variance of the N-acetyl aspartate concentrations. There were no associations between myo-inositol and N acetyl-aspartate or total creatine. Overall, mean N-acetyl-aspartate levels were below those reported by in vivo MRS studies of control subjects. CONCLUSIONS: These data suggest that decreased N-acetyl-aspartate in mesial temporal lobe epilepsy reflects altered mitochondrial metabolism, not merely neuron loss or gliosis. It is hypothesized that the altered N-acetyl-aspartate and creatine metabolism could reflect mitochondrial dysfunction or proliferation of immature glial cells that could contribute to the epileptogenic state. PMID- 12771257 TI - Survival after pulmonary edema due to enterovirus 71 encephalitis. AB - BACKGROUND: A distinctive pattern of enterovirus 71 (EV71) infection, characterized by fever, exanthem, acute pulmonary edema (PE), brainstem encephalitis, and flaccid paresis, affects infants and young children. Most die rapidly owing to respiratory failure and fulminant PE. METHOD: The authors report short- and long-term outcome of six survivors of the acute illness. RESULTS: In the context of acute PE and widespread weakness, recognition of the underlying neurologic disorder was facilitated by the distinctive pattern of MRI signal abnormalities in posterior pons and medulla. EV71-specific PCR of clinical samples helped confirm the diagnosis. Acute PE was managed with mechanical ventilation, afterload reduction, and inotrope support, and resolved completely over days. One patient with minimal neurologic recovery died 9 weeks after disease onset. The other patients have residual neurologic dysfunction, varying from subtle monoparesis to severe bulbar dysfunction, central and peripheral respiratory failure, and flaccid quadriparesis. Faster neurologic recovery was associated with less long-term deficit. Long-term outcome was similar in patients treated with and without pleconaril or IV immunoglobulin. Three long-term survivors treated with IV corticosteroids had less severe long-term neurologic disability than two not treated with steroids. CONCLUSION: Acute pulmonary edema and encephalomyelitis occurs with EV71 infection in infants. Long-term neurologic outcome varied from minor, focal weakness to profound, global motor dysfunction with respiratory failure. PMID- 12771258 TI - Altered cerebral vasoregulation in hypertension and stroke. AB - BACKGROUND: Autoregulation of blood flow that maintains steady perfusion over the range of systemic blood pressure is compromised by stroke. OBJECTIVES: To determine whether cerebral vasoregulation is impaired in stroke during orthostatic stress. METHODS: Subjects included 30 control subjects, 30 with hypertension, and 20 with minor stroke and were studied using transcranial Doppler. Bilateral blood flow velocities (BFVs) from middle cerebral arteries, heart rate, blood pressure (BP), and CO2 were obtained during hyperventilation and CO2 rebreathing during supine rest and tilt at 80 degrees. Side-to-side BFV difference, vasomotor range (VMR), and cerebrovascular resistance (CVR) were calculated during normo-, hypo-, and hypercapnia. RESULTS: Mean BFVs were similar between groups in supine position but differed during tilt. BFV diminished on the stroke side during tilt with hyperventilation and CO2 rebreathing (p < 0.0001). CVR increased (p < 0.0001) and VMR decreased (p < 0.01) on the stroke side. Vasoregulation was preserved on the normal side. BFV asymmetry differentiated patients with stroke from the other groups (p < 0.0001). BFV difference between the normal vs stroke side was the largest in stroke-normotensive (n = 7) compared with stroke-hypertensive (n = 13) patients and the two other groups (p < 0.0001). BFV asymmetry in stroke was associated with lower orthostatic BP (p < 0.0001). CONCLUSIONS: Cerebral vasoregulation is impaired with minor stroke, and cerebral blood flow depends on blood pressure. Decline of blood flow velocities during orthostatic stress may pose a risk of silent hypoperfusion. PMID- 12771260 TI - FDG-PET in paraneoplastic neuropathy. PMID- 12771259 TI - A novel form of pontocerebellar hypoplasia maps to chromosome 7q11-21. AB - OBJECTIVE: To describe a novel form of pontocerebellar hypoplasia (PCH) and map its genetic locus. BACKGROUND: PCH is a heterogeneous group of disorders that are characterized by abnormally small cerebellum and brainstem. Autosomal recessive inheritance has been implied in many cases, but no genetic loci have been mapped to date. METHODS: The authors studied a consanguineous family from the Sultanate of Oman with three siblings with a novel form of PCH. The authors performed clinical studies and linkage analysis of this pedigree. RESULTS: The clinical features of the affected children include developmental delay, progressive microcephaly with brachycephaly, seizures during the first year of life, hypotonia with hyperreflexia, short stature, and optic atrophy. Imaging studies showed a small pons and cerebellum, prominent sulci and lateral ventricles, and decreased cerebral white matter volume. A lack of dyskinesias distinguishes this pedigree from PCH type 2. Genetic studies of this family revealed evidence of significant linkage to chromosome 7q11-21 (maximum multipoint lod score 3.23). CONCLUSIONS: This pedigree represents a novel form of autosomal recessive PCH, which the authors propose to call cerebellar atrophy with progressive microcephaly (CLAM). This disorder maps to chromosome 7q11-21, and this locus was named CLAM. This report represents the first identification of a genetic locus for PCH. PMID- 12771261 TI - Direct access of drugs to the human brain after intranasal drug administration? AB - It has been suggested that intranasal (IN) drug delivery could be used to administer drugs directly to the brain, bypassing the blood-brain barrier. Conclusive evidence of this proposed route of drug transport has not been observed by IN-IV comparison. In eight neurosurgery patients with a CSF drain, the uptake in CSF and plasma after IN and IV drug administration was compared. No evidence of direct access of the drugs from the nose to the CSF was found. PMID- 12771262 TI - Anti-ganglioside antibodies in idiopathic and hereditary cerebellar degeneration. AB - Cerebellar degeneration has been associated with gluten sensitivity and celiac disease. Patients with celiac disease may have neuropathy and antibodies to gangliosides. The authors investigated the presence of antiganglioside antibodies in 22 patients with hereditary and nonhereditary ataxia and found 64% reactive in a novel agglutination test. PMID- 12771263 TI - Prevalence of antigliadin antibodies in ataxia patients. AB - We determined antigliadin antibodies in 95 ataxia patients and 73 controls. Antibodies were positive in 8% of the controls, 19% of patients with sporadic ataxia, 8% of patients with recessive ataxia, and 15% of patients with dominant ataxia. Statistical comparison using chi(2) statistics did not reveal significant differences between the groups. Although we found a trend toward a higher prevalence of antigliadin antibodies in patients with sporadic ataxia and dominant ataxia, our data do not support an association of ataxia with antigliadin antibodies. PMID- 12771264 TI - Idebenone treatment in Friedreich patients: one-year-long randomized placebo controlled trial. AB - The authors carried out a 1-year, randomized, placebo-controlled trial of idebenone in 29 patients with Friedreich ataxia. They found significant reductions of interventricular septal thickness and left ventricular mass in the idebenone group vs the placebo group, with no improvement in other heart ultrasound measures or neurologic condition. The absolute cardiac changes were modest, but the findings suggest that larger trials should assess whether idebenone reduces ventricular hypertrophy in patients with Friedreich ataxia. PMID- 12771265 TI - Idebenone treatment in Friedreich's ataxia: neurological, cardiac, and biochemical monitoring. AB - The authors report 1-year prospective data on eight patients with Friedreich ataxia. Idebenone did not halt the progression of ataxia. At the end of therapy, cardiac ultrasound demonstrated significant reduction of cardiac hypertrophy in six of eight patients. Cardiac strain and strain rate imaging showed that the reduction of hypertrophy is preceded by an early and linear improvement in cardiac function. Idebenone reduced erythrocyte protoporphyrin IX levels in five of six patients with elevated baseline levels; however, changes did not consistently relate to cardiac improvement. PMID- 12771266 TI - Rates and predictors for relapse in medication overuse headache: a 1-year prospective study. AB - The authors prospectively studied 98 patients with medication overuse headache. The 1-year relapse rate was 38% but was lower for patients with migraine compared with tension-type headache (22% vs 73%, p < or = 0.002) and combination of migraine and tension-type headache (22% vs 77%, p < or = 0.0001). The rate was also lower for patients overusing triptans than ergots (19% vs 20%, NS) and analgesics (19% vs 58%, p < or = 0.001). The long-term success of withdrawal depends on the type of primary headache and the type of overused medication. PMID- 12771267 TI - Immediate reocclusion following a successful thrombolysis in acute stroke: a pilot study. AB - Immediate reocclusion after successful thrombolysis was angiographically examined in 29 consecutive stroke patients. The reocclusion, which was associated with a high-grade residual stenosis of initially perfused arteries, occurred in four patients. IV abciximab successfully dissolved the thrombus in all four patients. Rapid thrombotic reocclusion after successful thrombolysis is commonly observed in patients after an acute stroke. Administration of abciximab may lead to reperfusion of the reoccluded artery. PMID- 12771268 TI - Novel LGI1 mutation in a family with autosomal dominant partial epilepsy with auditory features. AB - Autosomal dominant partial epilepsy with auditory features (ADPEAF) is a rare idiopathic epilepsy syndrome caused by mutations in the leucine-rich, glioma inactivated 1 (LGI1) gene. The authors report that molecular genetic studies in seven affected family members identified a novel F318C substitution that alters a highly conserved residue in a predicted repeat domain of unknown function. This report suggests that this domain may participate in the development of the ADPEAF phenotype. PMID- 12771269 TI - Cardiac troponin levels following monitored epileptic seizures. AB - Cardiac arrhythmia associated with myocardial injury is a proposed mechanism for sudden unexplained death in epilepsy. The authors measured serial cardiac troponin levels in 11 patients after monitored seizures. Using highly sensitive assays and criteria, no troponin elevations were seen, indicating that myocardial injury does not occur during uncomplicated seizures. An elevation in postictal troponin elevations should suggest the presence of cardiac injury secondary to neurocardiogenic mechanisms or primary cardiac factors, prompting further evaluation. PMID- 12771270 TI - Transient obstructive sleep apnea and asystole in association with presumed viral encephalopathy. AB - Evidence suggests that untreated obstructive sleep apnea (OSA) can lead to hypertension, cardiovascular disease, and stroke. Conversely, the systemic effects of a wide variety of critical illnesses can lead to CNS dysfunction, which can precipitate respiratory failure. Reported is a patient in whom an acute encephalopathy may have been responsible for transient OSA. PMID- 12771271 TI - Post-irradiation polyradiculopathy mimics leptomeningeal tumor on MRI. AB - Three patients with a remote history of Hodgkin's disease treated with total or subtotal lymphoid radiation 17 to 24 years earlier developed lumbosacral polyradiculopathy associated with nodular meningeal enhancement of the conus medullaris and cauda equina on MRI. None had evidence of recurrent Hodgkin's disease or second malignancy, and the MRI findings may be sequelae of radiation therapy. PMID- 12771272 TI - Contrasting effects of IFNbeta and IVIG in children with central and peripheral demyelination. AB - Reported are three children with MS who responded dramatically to interferon-beta (IFNbeta) therapy. While on immunomodulatory therapy, they developed chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) that responded to IV immunoglobulin (IVIG) administration. These cases emphasize two interesting observations: 1) IFNbeta treatment did not prevent development of CIDP; 2) CIDP in the context of MS responded to IVIG, even though IVIG had no therapeutic effect on the central demyelinating disease. PMID- 12771273 TI - Nodulus infarction mimicking acute peripheral vestibulopathy. AB - The authors report two patients with cerebellar infarctions in the territory of the medial branch of the posterior inferior cerebellar artery who had vertigo, spontaneous ipsilesional nystagmus, and contralesional truncal lateropulsion. Although one of the two patients had slight dysmetria, overall signs closely mimicked those of acute peripheral vestibulopathy. The authors suggest that interruption of nodulouvular inhibitory projections to vestibular nuclei may account for the vestibular signs. PMID- 12771274 TI - Canavan's leukodystrophy is associated with defects in cochlear neurodevelopment and deafness. AB - The authors present the temporal bone histopathology of two siblings (4 months old and 6 months old at autopsy) with Canavan's disease, an autosomal recessive leukodystrophy that is variably associated with sensorineural hearing loss. The histopathology demonstrated bilateral absence of the organ of Corti throughout the apical and basal cochlea and mild secondary atrophy of the spiral ganglia neurons. The vestibular end organs and ganglia were normal. These findings implicate a role of aminoacylase II in the neurodevelopment of the organ of Corti. PMID- 12771275 TI - Palmaris brevis spasm: an occupational syndrome. AB - Described are five patients who developed palmaris brevis (PB) spasm syndrome following prolonged use of a computer mouse and keyboard. Electromyography showed spontaneous activity characterized by irregular discharges of single motor unit potentials and myokymia from PB muscle that did not disappear after ulnar nerve block at the wrist, suggesting a distal lesion. PMID- 12771276 TI - Art and the brain: the influence of frontotemporal dementia on an accomplished artist. AB - A talented artist developed a progressive aphasia syndrome associated with frontotemporal dementia (FTD). As her disease progressed, language and executive skills declined, but her paintings became freer and more original. She demonstrates that artistic development can occur in the setting of language dominant types of FTD. The study of artistic development in the setting of FTD suggests that language is not required for, and may even inhibit, certain types of visual creativity. PMID- 12771277 TI - Treatment of slow-channel congenital myasthenic syndrome with fluoxetine. AB - The authors found that fluoxetine significantly shortens at 5 microM/L and nearly normalizes at 10 microM/L the prolonged opening bursts of slow-channel congenital myasthenic syndrome (SCCMS) acetylcholine receptors (AChR) expressed in fibroblasts. Prompted by this observation, they treated two SCCMS patients allergic to quinidine with up to 80 to 120 mg of fluoxetine per day over 3 years (serum fluoxetine + norfluoxetine levels 8 to 11 microM/L). Both patients showed marked subjective and objective improvement by quantitative muscle strength testing and electromyography. PMID- 12771278 TI - Differential activation on fMRI of monozygotic twins discordant for AD. AB - This is the first report of fMRI in monozygotic twins discordant for AD. FMRI brain activation patterns were examined during visuospatial and verbal working memory tasks. The affected twin had greater parietal involvement bilaterally during both working memory tasks and reduced left dorsolateral prefrontal cortex activity on the visuospatial memory task. Thus, fMRI may identify additional brain regions recruited in patients with AD to perform a given cognitive task. PMID- 12771279 TI - Acute cerebellitis with hydrocephalus. PMID- 12771280 TI - Enteropathy-associated T-cell lymphoma with initial manifestation in the CNS. PMID- 12771281 TI - A family with Campylobacter enteritis: anti-GD1a antibody with/without Guillain Barre syndrome. PMID- 12771282 TI - Does CMT1A homozygosity cause more severe disease with root hypertrophy and higher CSF proteins? PMID- 12771283 TI - Hemicrania horologica ("clock-like hemicrania"). PMID- 12771284 TI - Reversible hypoglycemic coma despite bilateral absence of the median nerve N20 evoked potential. PMID- 12771285 TI - A child with spinal cord AVM presenting with raised intracranial pressure. PMID- 12771286 TI - Ophthalmomyiasis-like pigmentary retinopathy in ALS/PDC in the Kii peninsula of Japan. PMID- 12771287 TI - Ophthalmoplegia in Powassan encephalitis. PMID- 12771288 TI - Patient pages. Daily headaches due to medication overuse. PMID- 12771289 TI - Management of secondary hyperparathyroidism. Proceedings of an international symposium on secondary hyperparathyroidism held by the Japanese Society for Parathyroid Intervention. August 17, 2002. Tokyo, Japan. PMID- 12771290 TI - Medical management of secondary hyperparathyroidism in chronic renal failure. AB - Abnormalities in calcium and phosphorus metabolism are common, and metabolic bone disease develops often in patients with chronic renal failure (CRF). Effective clinical management includes measures to control phosphorus retention and prevent hyperphosphataemia, to maintain serum calcium concentrations within the normal range and to prevent excess parathyroid hormone (PTH) secretion by the judicious use of vitamin D sterols. Certain of these interventions appear to increase the risk of soft tissue and vascular calcification in patients with end-stage renal disease (ESRD), changes that may contribute to the development of cardiovascular disease. Current therapeutic approaches are thus being re-evaluated in an effort to limit these risks. Despite the importance of controlling phosphorus retention and preventing hyperphosphataemia in patients with CRF, current management strategies often are inadequate, particularly in those ingesting diets containing adequate amounts of protein. Results from clinical trials using daily haemodialysis strongly suggest that thrice-weekly haemodialysis regimens are only marginally adequate for achieving weekly phosphorus balance in many patients with ESRD. The safety of large oral doses of calcium as a phosphate-binding agent in patients with ESRD has also been questioned because excess amounts of calcium that are absorbed from the gastrointestinal tract may lead to ongoing calcium retention in those with little or no residual renal function. Arterial calcification and cardiac valve calcification are two serious complications that adversely affect cardiovascular haemodynamics. The use of large, often supraphysiological, doses of calcitriol or other vitamin D sterols to treat secondary hyperparathyroidism may aggravate hypercalcaemia and hyperphosphataemia, further increasing the risk of soft tissue and vascular calcification. Phosphate-binding agents that do not contain calcium, new vitamin D analogues and calcimimetic compounds offer new therapeutic alternatives for managing renal osteodystrophy. The integration of these novel agents into existing treatment regimens may provide safer and more effective methods for controlling secondary hyperparathyroidism and renal bone disease, while limiting the risks of soft tissue and vascular calcification in patients with CRF. PMID- 12771291 TI - Parathyroid cell growth in patients with advanced secondary hyperparathyroidism: vitamin D receptor and cyclin-dependent kinase inhibitors, p21 and p27. AB - Uraemic patients with advanced secondary hyperparathyroidism (2HPT) have nodular hyperplastic glands with a decreased vitamin D receptor (VDR) density. Previous studies have shown that nodular hyperplasia expressed a significantly lower VDR density as compared with diffuse hyperplasia, and the VDR density negatively correlated with both the glandular weight and the marker of cell proliferation. However, the mechanism by which the decreased VDR density leads to parathyroid cell proliferation remains unclear. In the myelomonocytic cell line, active vitamin D(3) is known to activate the transcription of both p21 and p27, cyclin dependent kinase inhibitors (CDKIs), regulating the transition from the G(1) to the S phase of the cell cycle, in a VDR-dependent manner. Moreover, the overexpression of p21 and p27 inhibits cell proliferation. In order to elucidate the mechanism of parathyroid cell proliferation, the expression of CDKIs, p21 and p27, and the VDR was analysed immunohistochemically, and compared among nodular and diffuse hyperplastic parathyroid glands, and histologically normal parathyroid glands. The VDR expression in nodular hyperplasias was significantly decreased compared with either diffuse hyperplasias or normal parathyroid glands. The expression of both p21 and p27 was also significantly lower in nodular hyperplasias than in diffuse hyperplasias or normal parathyroid glands. Sections of parathyroid glands with a high expression of nuclear VDR highly expressed both p21 and p27. In nodular hyperplasias, the expression of both p21 and p27 correlated either positively with the nuclear VDR expression or inversely with the glandular weight. Therefore, the reduced expression of p21 and p27, being VDR dependent, is a major pathogenic factor for nodular parathyroid gland growth in advanced 2HPT. PMID- 12771292 TI - Control of parathyroid cell growth by calcimimetics. AB - Parathyroid cell hyperplasia is commonly observed in patients with chronic renal insufficiency and largely accounts for refractory secondary hyperparathyroidism. Calcimimetics are newly synthesized compounds that activate a calcium receptor on the parathyroid cell and can suppress parathyroid hormone secretion. The calcimimetic compound AMG 073 has been examined in clinical trials, and the data obtained so far demonstrate that the compound can lower the circulating levels of parathyroid hormone and calcium-phosphorus product in patients with secondary hyperparathyroidism. Furthermore, experimental evidence indicates that calcimimetics have the potential to inhibit parathyroid cell proliferation and block the progression of parathyroid hyperplasia. These beneficial effects, especially the potential to control parathyroid cell proliferation, would place calcimimetics among the essential therapeutic agents for treating secondary hyperparathyroidism. PMID- 12771293 TI - Adenovirus-mediated functional gene transfer into parathyroid cells in vivo and in vitro. AB - Chronic renal failure patients usually develop secondary hyperparathyroidism and, as the disease progresses, there is a decrease in the number of vitamin D and calcium-sensing receptors (CaRs) in the parathyroid glands. Parathyroid cell function can be controlled if a functional gene is transferred into these cells using an adenovirus vector. Vitamin D or CaR genes transferred by the infected adenovirus vector induced a reduction in parathyroid hormone secretion. These results suggest that adenovirus-mediated gene transfer is a useful technique for control of parathyroid cell function. PMID- 12771294 TI - Role of parathyroid intervention in the management of secondary hyperparathyroidism. AB - Measurement of the size of the parathyroid glands is mandatory for the selection of the optimal therapy for secondary hyperparathyroidism. Surgical parathyroidectomy or parathyroid intervention is indicated for patients with nodular hyperplasia as this form is seldom responsive to medical therapy. Selection of the intervention should be determined by the number, size and location of the parathyroid glands with nodular hyperplasia, including ectopic glands. PMID- 12771295 TI - Ultrasonographic diagnosis of parathyroid glands and percutaneous ethanol injection therapy. AB - The first choice for imaging diagnosis of parathyroid gland (PTG) abnormalities is ultrasonography with a high-frequency probe. The patient must be positioned correctly when performing either imaging or percutaneous ethanol injection (PEIT) of the PTG. The enlarged PTGs are examined by ultrasonic tomography using 3D measurements, and it is important to evaluate blood flow; the PTGs are hypervascular in comparison with a nodular lesion of the thyroid. Based on the 3D data, the estimated volume of the gland is calculated (a x b x c x pi/6), from which the volume of ethanol to be injected is determined. The operator performing the puncture should be skilled in interventional ultrasonography, including needle biopsy of superficial organs, because when the PTG enlargement is advanced, ethanol must be injected in several steps while checking for residual blood flow. After locating the tip of the needle by ultrasonography, the ethanol is injected, the jet echoes that spread from the tip of the needle are confirmed, and when there is no leakage from the gland, a volume of ethanol corresponding to approximately 50% of the volume of the enlarged gland is injected. If residual blood flow is observed, additional ethanol is injected at the site. After completion of PEIT of the PTG, colour Doppler imaging is repeated and, if there is no blood flow, then the procedure is considered to be successful. PMID- 12771296 TI - Guidelines for percutaneous ethanol injection therapy of the parathyroid glands in chronic dialysis patients. AB - Percutaneous ethanol injection therapy (PEIT) of the parathyroid was originally introduced as an alternative to surgical parathyroidectomy. After the recent elucidation of the pathogenesis of parathyroid hyperplasia in uraemia, 'selective PEIT of the parathyroid glands' was developed, in which enlarged parathyroid glands with nodular hyperplasia are 'selectively' destroyed by ethanol injection, and other glands with diffuse hyperplasia are then managed by medical therapy. The 'Guidelines for percutaneous ethanol injection therapy of the parathyroid glands in chronic dialysis patients' proposed by the Japanese Society for Parathyroid Intervention are presented, including indications, techniques, and post-PEIT management. These guidelines also apply to direct injection therapy using drugs other than ethanol, such as calcitriol and 22-oxacalcitriol. PMID- 12771297 TI - Evaluation of blood supply to the parathyroid glands in secondary hyperparathyroidism compared with histopathology. AB - BACKGROUND: In chronic renal failure patients, the parathyroid glands progress from diffuse hyperplasia to nodular hyperplasia, and it is important to distinguish between these as the latter form is more aggressive. This progress can be confirmed histologically, but the present study aimed to determine whether the different types of hyperplasia could be distinguished by power-Doppler ultrasonography (US). METHODS: Twenty-one consecutive renal failure patients were scheduled to undergo parathyroidectomy (PTx). Of 70 resected parathyroid glands, 63 were assessed by pre-operative power-Doppler US, classified into four groups based on the flow signal pattern and then correlated with the post-operative histopathology. RESULTS: With power-Doppler US imaging, 60.0% of glands without a signal inside the gland were diagnosed as diffuse hyperplasia or diffuse hyperplasia with early nodularity. Of glands with in-gland signals, 83.7% were nodular or had a single nodule typical of nodular hyperplasia. Even when the focus was on parathyroid glands weighing 2000 mm(3)) appeared to be resistant to this intervention and an intrathoracic parathyroid gland was found in a non responsive case. None of the patients had any severe complications, such as nerve palsy or massive haemorrhage. CONCLUSION: This new approach to the control of 2HPT is recommended as an alternative pharmacological parathyroidectomy to surgical therapy. PMID- 12771299 TI - Effect of percutaneous calcitriol injection therapy on secondary hyperparathyroidism in uraemic patients. AB - BACKGROUND: The impetus to develop percutaneous calcitriol injection therapy (PCIT) was the lack of therapeutic tools to treat secondary hyperparathyroidism (2HPT) resistant to medical therapy. METHODS: Nine dialysis patients resistant to intravenous calcitriol or calcitriol analogues underwent daily PCIT 5-10 times consecutively. The PCIT involved the injection of a volume of calcitriol equal to that of the enlarged parathyroid glands (PTGs) under ultrasonographic guidance. All patients had follow-up intravenous calcitriol after PCIT. RESULTS: The serum intact PTH concentration was markedly reduced following PCIT and was maintained for 12 weeks with intravenous calcitriol without significant changes in serum adjusted calcium and phosphorus concentrations. All patients tolerated PCIT without serious adverse events. Serum bone alkaline phosphatase concentrations and the volume of the enlarged PTGs were also significantly reduced. CONCLUSION: PCIT is a safe and effective treatment, which may also suppress parathyroid hyperplasia and improve bone turnover for refractory 2HPT. PMID- 12771301 TI - Surgical verification of percutaneous maxacalcitol injection therapy on enlarged parathyroid glands in chronic dialysis patients. AB - Selective percutaneous ethanol injection therapy (PEIT) has been used to control parathyroid function in patients with secondary hyperparathyroidism (2HPT) when one or more parathyroid gland (PTG) progresses to the nodular hyperplasia stage. However, PEIT can have adverse side effects, such as nerve paralysis and adhesion, because the ethanol is destructive. Intraparathyroid injection of a vitamin D analogue has been designed as a treatment to control parathyroid function without destruction of the PTG or causing adhesions to the surrounding tissue, and the present study aimed to verify the effect of percutaneous maxacalcitol (22-oxacalcitriol) as the vitamin D analogue. The study group comprised two haemodialysis patients who needed parathyroidectomy for uncontrolled 2HPT with a maximal PTG diameter >20 mm. The treatment began with an ultrasonographically guided injection of 10 microg of maxacalcitol solution into the largest PTG and, 1 week later, parathyroidectomy was performed to examine the effect of the maxacalcitol injection both macroscopically and microscopically. The injected glands were swollen and inflamed, and adhesions made it difficult to remove them. There was macroscopic and microscopic evidence of haemorrhagic necrosis and adhesions to the surrounding tissue. Direct vitamin D analogue injection should not be performed as a primary treatment option because the adverse side effects are not overcome by this technique. PMID- 12771302 TI - Time course of change in calcium x phosphorus product after percutaneous ethanol injection therapy. AB - BACKGROUND: Percutaneous ethanol injection therapy (PEIT) effectively suppresses PTH secretion, but the change in the serum calcium and phosphorus product (Ca x P) after PEIT has not been fully evaluated. METHODS: Twenty-seven haemodialysis patients with severe secondary hyperparathyroidism (2HPT) were divided into two groups according to their intact PTH (i-PTH) concentrations 6 months after PEIT: (i). effective (E) group, i-PTH concentration <360 pg/ml; and (ii). non-effective (N) group i-PTH > or = 360 pg/ml. The changes in serum calcium and phosphorus concentrations and the Ca x P were recorded for the following 2 years under post PEIT medical treatment with oral calcitriol or intravenous 22-oxacalcitriol (OCT). RESULT: In the E group, the i-PTH concentrations decreased to <300 pg/ml 1 year after PEIT (801+/-302 to 280+/-134 pg/ml), then increased to 435+/-201 pg/ml at 2 years. Serum calcium concentration did not show any significant change except for a transient reduction at 1 month after PEIT. The Ca x P decreased for 1 year (from 66.3+/-15.3 to 56.2+/-10.3 mg(2)/dl(2); P<0.05), in agreement with the course of phosphorus concentration, and continued to be <60 mg(2)/dl(2) up to 2 years after PEIT. The Ca x P tended to decrease more with OCT than oral calcitriol. In the N group, calcium and Ca x P increased significantly at 6 months after PEIT and remained at a high value. CONCLUSION: Treatment with PEIT suppresses serum PTH concentration as well as Ca x P in the long term. PMID- 12771303 TI - Long-term (3 years) prognosis of parathyroid function in chronic dialysis patients after percutaneous ethanol injection therapy guided by colour Doppler ultrasonography. AB - BACKGROUND: Secondary hyperparathyroidism (2HPT) is one of the most important complications in chronic dialysis (CD) patients. Percutaneous ethanol injection therapy (PEIT) of the parathyroid glands was introduced initially as an alternative treatment to parathyroidectomy and, with the technical progress of parathyroid imaging, it has now become a useful adjunct to medical therapy. The present study examined the possibility of maintaining parathyroid function in the long term (3 years) after PEIT. METHOD: PEIT, guided by power-Doppler flow mapping, was performed in 33 CD patients with severe 2HPT, and all glands >5.0 mm in diameter were destroyed. RESULTS: All patients showed a decline in the serum intact parathyroid hormone (i-PTH) concentration, on average from 695.5 to 248.0 pg/ml, after 1 year. After 3 years, the i-PTH concentration was controlled at <300 pg/ml in 85% of patients. The mean serum alkaline phosphatase (ALP) concentration also decreased from 322.7 to 154.4 IU/l after 1 year. In 76% of patients, ALP was maintained within the normal range (between 76 and 260 IU/l) at 3 years. Patients were classified into four groups according to the number of parathyroid glands detected by ultrasonography: one gland in group 1, two in group 2, three in group 3, and four in group 4. At 3 years after PEIT, i-PTH was controlled at <300 pg/ml in 100, 79, 83 and 82% of the patients in groups 1-4, respectively. CONCLUSIONS: It was possible to maintain long-term parathyroid function after PEIT in patients with 2HPT by using medical therapy, such as oral calcitriol pulse therapy and additional PEIT. PMID- 12771304 TI - Intervention for recurrent secondary hyperparathyroidism from a residual parathyroid gland. AB - There is a significant recurrence rate of secondary (renal) hyperparathyroidism after total parathyroidectomy (PTx) with forearm autograft. The lesions responsible for recurrent hyperparathyroidism are mainly the parathyroid autografts, but in some cases there are previously undetected residual or ectopic parathyroid glands. In Kojinkai hospitals, 155 haemodialysis out-patients had total PTx and forearm autograft for severe renal hyperparathyroidism and, during the past 18 years, 40 of them developed recurrent or persistent renal hyperparathyroidism. Five patients were treated by percutaneous ethanol injection therapy (PEIT): four patients had residual parathyroid glands and one patient had an ectopic parathyroid gland. The results of PEIT depended on the functioning of the parathyroid autografts. In two patients with non-functioning autografts, the effect of PEIT was remarkable; both showed 'hungry bone' syndrome and became hypoparathyroid. In the three patients with functioning autografts, the clinical course after PEIT was mild, but resection of the autograft was required in one patient. When an echo-guided approach is possible, PEIT for residual parathyroid glands is an effective intervention for the management of recurrent renal hyperparathyroidism; however, there is a risk of hypoparathyroidism in patients with non-functioning parathyroid autografts. As parathyroid autografts consist of multiple nodules, echo-guided injection of ethanol or calcitriol to each nodule is almost impossible and therefore resection of the autograft is indicated for autograft-dependent recurrent renal hyperparathyroidism. PMID- 12771305 TI - Re-operation is frequently required when parathyroid glands remain after initial parathyroidectomy for advanced secondary hyperparathyroidism in uraemic patients. AB - BACKGROUND: Parathyroidectomy (PTx) is the most successful treatment for advanced secondary hyperparathyroidism (2HPT) not responsive to medical treatment. However, persistent HPT remains problematic after PTx if some glands remain. The clinical course in patients with persistent 2HPT was evaluated to clarify the risk for re-operation after PTx. METHODS: Between March 1981 and December 2001, initial total PTx with forearm autograft were performed in 1156 uraemic patients. Persistent HPT cases were defined as those in which the lowest post-operative intact parathyroid hormone (i-PTH) concentration was >60 pg/ml, and patients were classified into groups A, B and C, with i-PTH concentrations of >or=500, 300-500 and 60-300 pg/ml, respectively. These patients were followed for 7-234 months after PTx. RESULTS: Persistent HPT was identified in 49/1156 patients (4.2%), with nine cases in group A, 10 in group B and 30 in group C. Re-operation was required in 21/49 (42.8%) cases, and in seven of these the last i-PTH concentration was >or=500 pg/ml. All cases in group A required re-operation. In group C, 11/30 (36.7%) patients required re-operation. The missed glands removed at re-operation were supernumerary in 14 cases, and located in the mediastinum in 13 cases. The frequency of advanced HPT and re-operation was not negligible. CONCLUSIONS: To prevent persistent 2HPT, all parathyroid glands must be found and resected during the initial operation. Even if small parathyroid glands remain, there is a risk of progression. Complete PTx is the first treatment choice for advanced 2HPT. PMID- 12771306 TI - Long-term prognosis of parathyroid function for chronic dialysis patients after minimally invasive radioguided parathyroidectomy (MIRP). AB - BACKGROUND: Minimally invasive radioguided parathyroidectomy (MIRP) for primary hyperparathyroidism for one gland, located by scanning with technetium 99m labelled sestamibi (MIBI), has been performed. Total parathyroidectomy with autotransplantation or percutaneous ethanol injection therapy (PEIT) for severe secondary hyperparathyroidism (2HPT) has also been performed. METHODS: The present study examined the possibility of maintaining parathyroid function within a target range [intact parathyroid hormone (i-PTH) 2.2% (P < 0.055). Alternate explanations for this shift were deemed improbable. These findings show that FMNP goals were advanced through a coordinated, collaborative initiative with activities at state and local levels, resulting in increased utilization of FMNP benefits by WIC participants and increased income to local farmers. PMID- 12771328 TI - Fermentable fiber reduces recovery time and improves intestinal function in piglets following Salmonella typhimurium infection. AB - Diarrhea is a leading cause of morbidity and mortality in infants. The addition of fiber to infant formulas reduces recovery time following pathogenic infection in infants > 6 mo old, but effects on neonates are unknown. The hypothesis that fermentable fiber reduces infection-associated symptoms and enhances intestinal structure and function in neonates was examined. Piglets (2 d old) were randomly assigned to receive formula alone (control) or formula containing methylcellulose (MCEL), soy polysaccharides (SPS) or fructooligosaccharides (FOS) for 14 d. On d 7, piglets were further randomly assigned to receive an oral gavage of Salmonella typhimurium or serve as noninfected controls. S. typhimurium infection produced diarrhea in controls and MCEL groups, but not in the SPS and FOS groups. Postinfection physical activity was lower (P = 0.0001) in the controls than in all other groups. Ileal lactase activity was reduced (P < 0.05) following infection in the control group but not in the MCEL, SPS and FOS groups. Ileal mucosal barrier function, measured as resistance, was impaired by infection (P < 0.05) in the control and SPS groups, but was unaltered in the jejunum and colon. Total ion transport and basal short-circuit current were higher (P < 0.05) in jejunum than in ileum and colon, irrespective of diet or infection. SPS and FOS increased (P < 0.05) ileal glutamine transport relative to piglets fed MCEL, irrespective of infection. Because fermentable fiber enhances intestinal function and reduces the severity of S. typhimurium infection-associated symptoms, it may be a cost-effective way in which to reduce the severity of pathogenic infection associated symptoms in infants. PMID- 12771329 TI - Chlorogenic acid bioavailability largely depends on its metabolism by the gut microflora in rats. AB - Chlorogenic acid, the ester of caffeic acid with quinic acid, is one of the most abundant polyphenols in the human diet with coffee, fruits and vegetables as its major sources. Its antioxidant and anticarcinogenic properties have been well established in animal studies. However, little is known about its gut absorption and metabolism. In the present work, four groups of rats (n = 8) were fed a diet supplemented with chlorogenic, caffeic or quinic acids (250 micromol/d) or an unsupplemented diet for 8 d. Parent compounds and their metabolites were estimated in urine (24-h collection) and plasma by HPLC-electrospray ionization tandem mass spectrometry. Significant differences in their levels were observed among the groups. The recovery of chlorogenic acid in urine was low (0.8%, mol/mol), and the total urinary excretion of caffeic acid liberated by hydrolysis of chlorogenic acid and its tissular methylated metabolites (ferulic and isoferulic acids) did not account for >0.5% (mol/mol) of the dose ingested. On the other hand, the metabolites of microbial origin, namely, m-coumaric acid and derivatives of phenylpropionic, benzoic and hippuric acids, represented the major compounds in both urine and plasma. Hippuric acid largely originated from the transformation of the quinic acid moiety, and all other metabolites from the caffeic acid moiety. These microbial metabolites accounted for 57.4% (mol/mol) of the chlorogenic acid intake. Such a high abundance of microbial metabolites shows that the bioavailability of chlorogenic acid depends largely on its metabolism by the gut microflora. Their potential importance in explaining the biological effects of dietary polyphenols is emphasized. PMID- 12771330 TI - Apple pectin and a polyphenol-rich apple concentrate are more effective together than separately on cecal fermentations and plasma lipids in rats. AB - To evaluate the effect of apple components on cecal fermentations and lipid metabolism, rats were fed diets containing 5 g/100 g apple pectin (PEC), 10 g/100 g high polyphenol freeze-dried apple (PL) or both (PEC + PL). The cecal pH was slightly acidic (6.49) only in rats fed the PEC + PL diet (controls, 7.02). The cecal short-chain fatty acid pool was enlarged by all the apple fractions, with a peak of 560 micromol in rats fed the PEC + PL diet compared with 189 micromol in controls. Butyrate concentrations were 2-fold greater in rats fed the PL diet than in controls. Substantial concentrations of galacturonate and succinate (approximately 40 mmol/L) were found in the cecum of rats fed the PEC diet and, to a lesser extent, the PEC + PL diet. The PEC + PL diet significantly lowered plasma cholesterol, whereas both the PL and PEC + PL diets lowered plasma triglycerides. Liver cholesterol and triglyceride concentrations were lower in rats fed the PEC and PEC + PL diets. Fecal bile acid excretion was markedly reduced, whereas sterol excretion was significantly increased by dietary PEC. Rats fed the PEC and PEC + PL diets also had lower apparent cholesterol absorption than controls (30 compared with 43%). In conclusion, apple pectin and the polyphenol-rich fraction were more effective when fed combined together than when fed separately on large intestine fermentations and lipid metabolism, suggesting interactions between fibers and polyphenols of apple. PMID- 12771331 TI - Weight gain in gonadectomized normal and lipoprotein lipase-deficient male domestic cats results from increased food intake and not decreased energy expenditure. AB - Gonadectomy predisposes domestic cats to undesired body weight gain and obesity. The disturbance responsible for this disregulation of energy balance has not been clearly identified. Energy intake and expenditure, body composition and plasma concentrations of leptin, insulin, glucose and triacylglycerol were determined during a 36-wk period in adult male (2-5 y) gonadectomized (n = 8) and intact (n = 8) normal cats and gonadectomized (n = 8) and intact (n = 8) lipoprotein lipase (LPL)-deficient cats. Cats were housed individually in temperature- and light controlled rooms and continuously provided a commercial dry-type diet. In normal and LPL-deficient cats, body weight increased (P < 0.05) after gonadectomy by 27 to 29%, mostly as a result of fat accretion. There was a rapid increase (P < 0.05) in food intake of approximately 12% after gonadectomy of normal and LPL deficient cats. The metabolic rate (kJ.kg(-1).d(-1)), determined in normal intact (319 +/- 20, n = 5) and gonadectomized (332 +/- 36, n = 5) cats, did not differ after gonadectomy. After gonadectomy, plasma concentrations of glucose and triacylglycerol did not change, whereas plasma insulin and leptin concentrations increased (P < 0.05), but not coincidentally with body weight gain. A stair-step increase in energy intake, and not decreased energy expenditure, appears to drive the weight gain associated with gonadectomy. Body fat mass appears to increase until the energy intake supports no further expansion. Adiposity signaling through insulin or leptin does not appear to mediate the energy intake effect. LPL deficiency did not preclude development of the overweight body condition. Therefore, gonadectomy-induced weight gain in cats is not a result of changed adipose LPL activity, as previously suggested. PMID- 12771332 TI - Colitis increases albumin synthesis at the expense of muscle protein synthesis in macronutrient-restricted piglets. AB - Our aim was to examine the effect of acute inflammation localized in the colon and early macronutrient restriction on protein synthesis in a piglet model. In a 2 x 2 factorial design, piglets (n = 32) were fed an adequate or macronutrient restricted diet with or without dextran sulfate-induced colitis for 7 d. The stable isotope tracer L-[5,5,5-(2)H(3)]leucine was infused to determine protein kinetics at the whole-body level and synthesis of tissue and plasma proteins. In the well-nourished state, colitis did not affect weight gain or protein kinetics except for an increase in albumin synthesis (P < 0.05). Macronutrient restriction alone caused a general slowing of protein metabolism including decreased weight gain (P < 0.0004), whole-body protein turnover (P < 0.0001), and liver (P < 0.01) and plasma protein (P < 0.03) synthesis. However, in the presence of macronutrient restriction, colitis compromised weight gain further (P < 0.02) and decreased muscle protein synthesis (P < 0.05) due to a redistribution of protein metabolism that supported enhanced synthesis of plasma proteins. The increased contribution of plasma protein synthesis to whole-body protein turnover was attributable mainly to increased synthesis of albumin (P < 0.006). Concentrations of plasma proteins were unaffected despite dramatic changes in their synthesis rates, thereby underestimating the effects of malnutrition and colitis on protein metabolism. Increased synthesis of plasma proteins, particularly the negative acute phase reactant albumin, compromises weight gain and muscle protein synthesis only when macronutrient intake is inadequate, underscoring the role of adequate nutrition in preventing growth impairment and muscle wasting in acute inflammation. These results suggest that the hypoalbuminemia of inflammatory bowel disease should not be attributed to decreased synthesis. PMID- 12771333 TI - Dietary vitamin C and folic acid supplementation ameliorates the detrimental effects of heat stress in Japanese quail. AB - We evaluated the effects of vitamin C (L-ascorbic acid) and folic acid supplementation on performance, carcass characteristics and concentrations of the oxidative stress markers [malondialdehyde (MDA), homocysteine], adrenocorticotropic hormone (ACTH), vitamins C, E, A, B-12 and folic acid, and mineral status in broiler Japanese quail (Coturnix coturnix japonica) exposed to high ambient temperature (34 degrees C, 8 h/d, 0900-1700 h). The birds (n = 150; 10-d-old) kept at 34 degrees C were fed a basal diet (HS group) or the basal diet supplemented with 250 mg of L-ascorbic acid/kg of diet (Vit C group), 1 mg of folic acid/kg of diet (FA group) or both (Vit C + FA group), whereas birds kept at 22 degrees C were fed the basal diet (TN group). Supplementing heat-stressed quail with vitamin C and folic acid improved performance compared to the HS group. Effects generally were greatest in quail supplemented with both. Although supplementation did not consistently restore concentrations to those of the TN group, it increased serum concentrations of the vitamins under study. Furthermore, serum and tissue MDA, homocysteine and ACTH concentrations were lower in the supplemented groups than in the heat-stressed controls. Retention of N, ash, Ca, P, Zn, Fe, Cu and Cr were highest in the Vit C + FA group and lowest in the HS group (P < 0.05). The results of the study indicate that vitamin C and folic acid supplementation attenuates the decline in performance and antioxidant status caused by heat stress. Such supplementation may offer protection against heat stress-related depression in performance of Japanese quail. PMID- 12771334 TI - Epsilon-polylysine inhibits pancreatic lipase activity and suppresses postprandial hypertriacylglyceridemia in rats. AB - Epsilon-polylysine (epsilon-PL) has been used as a food additive in Japan for many years. In this study, it inhibited human and porcine pancreatic lipase activity in substrate emulsions containing bile salts and phosphatidylcholine, in the concentration range of 10-1000 mg/L. At the same concentrations, it also destroyed the emulsifying activity, suggesting that lipase inhibitory activity and emulsion breakdown activity were associated. Epsilon-PL inhibited porcine pancreatic lipase activity and destroyed emulsion breakdown activity at 1000 mg/L in the substrate containing bile salts and phosphatidylcholine alone. Epsilon-PL did not inhibit lipase activity or affect emulsifying activity at 1000 mg/L in the substrates containing arabic gum and polyvinyl alcohol. A comparison of lipase inhibitory activity between epsilon-PL and three types of alpha-PL with differing polymerization rates was performed. The lipase inhibitory activity of epsilon-PL was not different from that of alpha-PL (44 lysine residues). Epsilon PL maintained its inhibitory activity after incubation with trypsin, alpha chymotrypsin and pepsin, whereas alpha-PL did not. The effect of epsilon-PL on postprandial hypertriacylglyceridemia was investigated in rats. The plasma triacylglycerol concentration in rats intragastrically administered > or =15 mg/kg of both fat emulsion and epsilon-PL was significantly lower at 2 and 3 h after administration than that in rats administered fat emulsion alone (P < 0.05). These results strongly suggest that epsilon-PL is able to suppress dietary fat absorption from the small intestine by inhibiting pancreatic lipase activity. PMID- 12771335 TI - Hesperidin, a citrus flavonoid, inhibits bone loss and decreases serum and hepatic lipids in ovariectomized mice. AB - The purpose of this study was to examine whether hesperidin inhibits bone loss in ovariectomized mice (OVX), an animal model of postmenopausal osteoporosis. Forty 8-wk-old female ddY mice were assigned to five groups: a sham-operated group fed the control diet (AIN-93G), an OVX group fed the control diet, an OVX+HesA group fed the control diet containing 0.5 g/100 g hesperidin, and an OVX+HesB group fed the control diet containing 0.7 g/100 g alpha-glucosylhesperidin and an OVX+17beta-estradiol (E(2)) group fed the control diet and administered 0.03 micro g E(2)/d with a mini-osmotic pump. After 4 wk, the mice were killed and blood, femoral, uterine and liver were sampled immediately. Hesperidin administration did not affect the uterine weight. In OVX mice, the bone mineral density of the femur was lower than in the sham group (P < 0.05) and this bone loss was significantly prevented by dietary hesperidin or alpha glucosylhesperidin. The Ca, P and Zn concentrations in the femur were significantly higher in the hesperidin-fed and E(2) groups than in the OVX group. Histomorphometric analyses showed that the trabecular bone volume and trabecular thickness in the femoral distal metaphysis were markedly decreased (P < 0.05) by OVX, and alpha-glucosylhesperidin significantly prevented this bone loss. Furthermore, hesperidin decreased the osteoclast number of the femoral metaphysis in OVX mice, as did E(2). Serum and hepatic lipids were lower in mice that consumed the hesperidin-containing diets (P < 0.05) than in the OVX group fed the control diet. These results suggest a possible role for citrus flavonoids in the prevention of lifestyle-related diseases because of their beneficial effects on bone and lipids. PMID- 12771336 TI - Adult cockatiels (Nymphicus hollandicus) at maintenance are more sensitive to diets containing excess vitamin A than to vitamin A-deficient diets. AB - The purpose of this experiment was to examine the physiological responses of adult cockatiels at maintenance to dietary vitamin A (VA) concentrations, and to identify concentrations associated with deficiency and toxicity. Adult cockatiels at maintenance (n = 22, 2-3 y of age) were fed a diet of 0, 600, 3000 or 30,000 microg VA/kg (0, 2000, 10,000 or 100,000 IU), and monitored for signs of VA deficiency or toxicity for up to 706 d. The analyzed diet concentrations were 0, 835, 2815 and 24,549 microg/kg, respectively. After 269 d, birds fed the 30,000 microg/kg VA diet had greater plasma retinal concentrations, markedly intensified vocalization patterns, pancreatitis and multifocal accumulation of lymphocytes in the lamina propria of the duodenum compared to birds fed the 600 microg/kg diet (P < 0.05). The 3000 microg/kg VA diet induced increased plasma retinol, splenic hemosiderosis and altered vocalization patterns (P < 0.05), although not as striking as those induced by the 30,000 microg/kg VA diet. The secondary antibody response was reduced after 225 d and vocalization patterns were altered in birds fed 0 microg/kg VA (P < 0.05), but after almost 2 y there were no changes in body condition, plasma retinol, organ pathology or classical signs of deficiency such as squamous metaplasia of nasal epithelia. Thus, adult cockatiels at maintenance were more susceptible to VA toxicity than to VA deficiency and concentrations > or = 3000 microg VA/kg diet can cause toxicity. It is possible that disturbances in VA nutrition contribute to the widespread incidence of behavioral problems reported in companion birds. PMID- 12771337 TI - Oligofructose protects against the hypertriglyceridemic and pro-oxidative effects of a high fructose diet in rats. AB - Recent findings indicate that in addition to its hyperlipemic effect, a high fructose diet has a pro-oxidant effect in rats compared with a starch-based diet. Oligofructose (OFS) has already been shown to decrease plasma lipids in rats. We assessed the impact of fructose on oxidative stress by supplementing a high fructose diet with OFS. Rats were fed either a high fructose diet or a starch based diet, with or without supplementation of 10 g/100 g oligofructose for 4 wk. Regardless of the type of carbohydrate, OFS in the diet produced an enlargement of the cecum and led to a significant increase in the SCFA cecum pool. Fructose feeding was associated with significantly higher insulin plasma concentrations (+63%) in the control groups, whereas insulin plasma concentrations did not differ in rats fed the fructose diet supplemented with OFS. Plasma leptin concentration was significantly lower (approximately 50%) in the OFS-supplemented fructose group compared with the other three groups. Fructose feeding in rats also significantly increased plasma (P < 0.001) and liver (P < 0.001) triglyceride (TG) concentrations and the addition of OFS prevented the TG accumulation induced by fructose in the liver (P < 0.05) and hyperlipemia (P < 0.05). OFS consumption prevented (P < 0.05) the lower plasma vitamin E/TG ratio in rats fed the fructose diet. Control rats fed the fructose diet had high plasma TBARS values compared with rats fed the starch diet, whereas TBARS values remained unchanged when rats were supplemented with OFS. Control rats fed the fructose diet had higher TBARS urine values and higher heart tissue susceptibility to peroxidation compared with rats fed the starch diet, and this effect was significantly reduced by OFS consumption. Further studies are required to identify the mechanisms underlying the protective effect of OFS against the pro-oxidant effect of fructose. However, the potential nutritional benefits of OFS supplementation in fructose-rich diets are suggested. PMID- 12771338 TI - Safety assessment of recombinant green fluorescent protein orally administered to weaned rats. AB - Several proposed biotechnological applications of green fluorescent protein (GFP) are likely to result in its introduction into the food supply of domestic animals and humans. We fed pure GFP and diets containing transgenic canola expressing GFP to young male rats for 26 d to evaluate the potential toxicity and allergenicity of GFP. Animals (n = 8 per group) were fed either AIN-93G (control), control diet plus 1.0 mg of purified GFP daily, modified control diet with 200 g/kg canola (Brassica rapa cv Westar), or control diet with 200 g/kg transgenic canola containing one of two levels of GFP. Ingestion of GFP did not affect growth, food intake, relative weight of intestine or other organs, or activities of hepatic enzymes in serum. Comparison of the amino acid sequence of GFP to known food allergens revealed that the greatest number of consecutive amino acid matches between GFP and any food allergen was four, suggesting the absence of common allergen epitopes. Moreover, GFP was rapidly degraded during simulated gastric digestion. These data indicate that GFP is a low allergenicity risk and provide preliminary indications that GFP is not likely to represent a health risk. PMID- 12771340 TI - Intermittent food deprivation improves cardiovascular and neuroendocrine responses to stress in rats. AB - Stressful events may trigger disease processes in many different organ systems, with the cardiovascular system being particularly vulnerable. Five-mo-old male rats had ad libitum (AL) access to food or were deprived of food every other day [intermittent food deprivation (IF)] for 6 mo, during which time their heart rate (HR), blood pressure (BP), physical activity and body temperature were measured by radiotelemetry under nonstress and stress (immobilization or cold-water swim) conditions. IF rats had significantly lower basal HR and BP, and significantly lower increases in HR and BP after exposures to the immobilization and swim stressors. Basal levels of adrenocorticotropic hormone (ACTH) and corticosterone were greater in the IF rats. However, in contrast to large stress-induced increases in ACTH, corticosterone and epinephrine levels in AL rats, increases in these hormones in response to repeated immobilization stress sessions were reduced or absent in IF rats. Nevertheless, the IF rats exhibited robust hypothalamic/pituitary and sympathetic neuroendocrine responses to a different stress (swim). The IF treatment improved glucose metabolism, as indicated by lower basal levels of circulating glucose and insulin, but with maintenance of glucose and insulin responses to stress. We concluded that improvements in cardiovascular risk factors and cardiovascular and neuroendocrine stress adaptation occur in response to IF. PMID- 12771339 TI - Dietary (n-3) polyunsaturated fatty acids remodel mouse T-cell lipid rafts. AB - In vitro evidence indicates that (n-3) polyunsaturated fatty acids (PUFA) suppress T-cell activation in part by displacing proteins from lipid rafts, specialized regions within the plasma membrane that play an important role in T cell signal transduction. However, the ability of (n-3) PUFA to influence membrane microdomains in vivo has not been examined to date. Therefore, we compared the effect of dietary (n-3) PUFA on raft (liquid ordered) vs. soluble (liquid disordered) microdomain phospholipid composition in mouse T cells. Mice were fed diets containing either 5 g/100 g corn oil (control) or 4 g/100 g fish oil [contains (n-3) PUFA] + 1 g/100 g corn oil for 14 d. Splenic T-cell lipid rafts were isolated by density gradient centrifugation. Raft sphingomyelin content (mol/100 mol) was decreased (P < 0.05) in T cells isolated from (n-3) PUFA-fed mice. Dietary (n-3) PUFA were selectively incorporated into T-cell raft and soluble membrane phospholipids. Phosphatidylserine and glycerophosphoethanolamine, which are highly localized to the inner cytoplasmic leaflet, were enriched to a greater extent with unsaturated fatty acids compared with sphingomyelin, phosphatidylinositol and glycerophosphocholine. These data indicate for the first time that dietary (n-3) PUFA differentially modulate T cell raft and soluble membrane phospholipid and fatty acyl composition. PMID- 12771341 TI - Variations in plasma lycopene and specific isomers over time in a cohort of U.S. men. AB - Epidemiologic and laboratory studies suggest a possible role for tomato products, a rich source of the carotenoid lycopene, in the prevention of certain cancers and cardiovascular disease. Lycopene is consumed primarily as the all-trans isomer, but the majority of lycopene in blood and tissue exists as a variety of cis-isomers. Specific isomers may be involved in different biological reactions, and patterns of isomers may provide insight into the risk or pathogenesis of disease processes. Total lycopene concentration and the concentrations of the cis and trans-lycopene isomers were measured by HPLC in plasma samples taken 3-4 y apart from 144 mostly nonsmoking male participants in the Health Professionals Follow-up Study. Correlations between plasma concentrations determined 3-4 y apart ranged from 0.55 (all-trans-isomer) to 0.70 (cis-isomer 5 -cis) (P < 0.001). For total lycopene, the correlation was 0.63 (P < 0.001). Total cis lycopene contributed approximately 67% of total lycopene (range 50-79%). At each time point, the various lycopene isomer concentrations were highly correlated with one another with Spearman correlation coefficients ranging from 0.90 to 0.99 (P < 0.001). Plasma concentrations of total lycopene and its most abundant isomers in samples taken 3-4 y apart were strongly correlated, indicating that dietary patterns and metabolic processes defining lycopene concentrations are stable over time. Because the patterns of lycopene isomers showed limited between person variability, our results suggest that measuring specific lycopene isomers in epidemiologic studies may not provide additional information beyond that provided by total lycopene concentration. Single plasma samples quantitating plasma lycopene are a valid predictor of long-term exposure for epidemiologic studies. PMID- 12771342 TI - Risk of human ovarian cancer is related to dietary intake of selected nutrients, phytochemicals and food groups. AB - Intakes of specific nutrients and food groups have been shown previously to be related to ovarian cancer risk, but no studies, to our knowledge, have emphasized the effect of phytochemical intakes on this cancer. We conducted a case-control study of diet and ovarian cancer in western New York involving 124 primary, histologically confirmed ovarian cancer cases and 696 population-based controls, frequency matched to cases on age and county of residence. Diet was assessed with a detailed food-frequency questionnaire. Nutrient and phytochemical intakes were calculated from published food composition data. The odds ratios (OR) and 95% CI for risk of ovarian cancer with each nutrient, phytochemical and food group were estimated with unconditional logistic regression adjusting for age, education, total months menstruating, difficulty becoming pregnant, oral contraceptive use, menopausal status and energy intake. Compared with women in the lowest quintile of intake, reduced risks were observed for women in the highest quintile of intake of dietary fiber (OR 0.43, 95% CI, 0.20-0.94), total carotenoids (OR 0.33, 95% CI, 0.16-0.68), stigmasterol (OR 0.42, 95% CI, 0.20-0.87), total lignans (OR 0.43, 95% CI, 0.21-0.85), vegetables (OR 0.47, 95% CI, 0.23-0.97) and poultry (OR 0.45, 95% CI, 0.22-0.92). These results support a protective effect on ovarian cancer of phytoestrogen intakes, and our results support the hypothesis that a plant-based diet may be important in reducing risks of hormone-related neoplasms. PMID- 12771343 TI - Adenoma growth stimulation by the trans-10, cis-12 isomer of conjugated linoleic acid (CLA) is associated with changes in mucosal NF-kappaB and cyclin D1 protein levels in the Min mouse. AB - Conjugated linoleic acid (CLA) is a term used to describe the different conjugated isomers of linoleic acid. CLA has been found to be anticarcinogenic in mammary cancer, but its effects on colon carcinogenesis are still inconclusive. In this study, the isomer-specific effects of the cis-9, trans-11 and trans-10, cis-12 CLA isomers were investigated in the Min mouse model for intestinal carcinogenesis. The Min mice (n = 10/group) were fed either an AIN-93G control diet or a diet containing 1 g/100 g cis-9, trans-11 or trans-10, cis-12 CLA for 8 wk. The number and size of adenomas were measured and the proteins from the small intestinal tissues extracted for immunoblotting analysis. The number of adenomas did not differ, but the size of the adenomas was greater in the distal part of the small intestine in mice fed the trans-10, cis-12 isomer than in controls (1.19 +/- 0.16 vs. 0.94 +/- 0.21 mm, mean +/- SD, P < 0.01). The same isomer caused an increase in lipid peroxidation, measured as urinary 8-iso-prostaglandin (PG)F(2alpha). Nuclear p65 protein of the mucosal tissue was not detectable in the trans-10, cis-12 group, which differed (P < 0.05) from the control group. Cyclin D1, a target for the nuclear factor (NF)-kappaB pathway, was elevated in the trans-10, cis-12 group compared with the control group (P < 0.01), but cyclooxygenase-2 levels were not higher. There was no difference in beta-catenin protein levels between the groups. The results indicate that the trans-10, cis-12 isomer of CLA can act as a cancer promoter in colon carcinogenesis possibly through pathways affecting NF-kappaB and cyclin D1. PMID- 12771344 TI - The National Nutrition Summit: history and continued commitment to the nutritional health of the U.S. population. AB - Over the past several decades, great progress has been made in developing national nutrition policies and research agendas that address numerous problems faced by various segments of our population. Despite the success of these initiatives and our understanding of the importance of nutrition throughout the life cycle, ongoing nutrition-related problems have been identified and still need to be adequately addressed through research and policy. The recognition of these various issues inspired the development of an agenda for the National Nutrition Summit held in May 2000, jointly sponsored by the U.S. Departments of Agriculture and of Health and Human Services. The major goal of the Summit was to forge partnerships among policy makers and program planners at the federal, state, and community levels to examine what accomplishments were made since 1969 and to identify existing gaps in the areas of food, nutrition and health. This review provides an overview of the purpose and overarching themes that emerged from the Summit, along with a review of additional resources from the Summit available on a website sponsored by the U.S. Departments of Health and Human Services and Agriculture. PMID- 12771345 TI - Dietary supplement use in women: current status and future directions introduction and conference summary. PMID- 12771346 TI - Folic acid supplements and fortification affect the risk for neural tube defects, vascular disease and cancer: evolving science. AB - Folic acid supplements reduce the risk of neural tube defects and may be associated with reduced risk for vascular disease and cancer. Research data from both observational and controlled intervention studies provide strong support for the existing public health policies related to folic acid and neural tube defects. However, educational efforts to promote daily intake of folic acid supplements by women of reproductive age have not, in most cases, resulted in increased supplement use. In contrast, food fortification appears to be associated with a reduction in neural tube defects in the United States and Canada but is not practiced universally. The potential for folic acid supplements to reduce the incidence and severity of vascular disease and cancer is the focus of major research efforts including ongoing intervention studies. PMID- 12771347 TI - Micronutrients and reproductive health issues: an international perspective. AB - Micronutrients may have a role in enhancing reproductive health of women living in the developing world. Two illustrative micronutrients, zinc and vitamin A, have received some attention in this regard. Numerous animal experiments and observational studies suggest the potential role of zinc deficiency in labor and delivery-related complications such as premature rupture of membrane, placental abruption, preterm labor and inefficient uterine contraction. These associations have not been confirmed in supplementation studies. Zinc does not appear to be a limiting factor in intrauterine growth in the developing world, contrary to some evidence of its suggested benefit among women residing in industrialized countries. One study in Nepal found that maternal vitamin A or beta-carotene supplementation reduces pregnancy-related mortality but not infant mortality. These findings are corroborated by observations of the significantly higher risk of mortality among night-blind women compared to non-night-blind women long after the termination of pregnancy and the resolution of night blindness. Maternal multiple micronutrient supplementation needs more careful evaluation before its use in large-scale programs. Two recent trials indicated that a prenatal multiple micronutrient supplement provides no added advantage over iron and folate in reducing outcomes such as low birth weight and probably no survival benefit. Data are also suggestive that adding zinc may negate the beneficial effect of iron and folic acid on birth weight. Research is needed to further our understanding of nutrient-nutrient interactions. PMID- 12771348 TI - Iron supplement use among women in the United States: science, policy and practice. AB - The use of iron supplements is an accepted treatment for nonhereditary anemia. The use of iron supplements as prophylaxis is more controversial. We estimated the proportion of persons who consumed supplements that contain iron among the following groups: nonpregnant, nonlactating adolescents, aged 14-18 y (n = 992); women aged 19-50 y (n = 5,062); women aged 51 y and older (n = 3,593); pregnant women (n = 295); and lactating women (n = 97) using data from the National Health and Nutrition Examination Survey, 1988-1994. We found that the proportion (% +/- SE) of U.S. women consuming supplements containing iron in the previous month was 9 +/- 2% among nonpregnant, nonlactating adolescents; 23 +/- 1% among women aged 19 y and older; 72 +/- 4% among pregnant women; and 60 +/- 8% among lactating women. Low income women were less likely to consume supplements containing iron. Minority women were less likely to consume supplements containing iron in all groups except adolescents. Among consumers of supplements that contain iron, the median intake of iron was 11 mg/d among nonpregnant adolescents, approximately 17 mg/d among nonpregnant women, 58 mg/d among pregnant women and 57 mg/d among lactating women. Use of supplements that contain iron was associated with a significantly reduced prevalence of iron deficiency among women 19-50 y but not among other groups. Groups at highest risk of iron deficiency (e.g., low income and minority women) are often least likely to consume supplements containing iron, suggesting that supplement use is unrelated to actual need. PMID- 12771349 TI - Environmental influences: factors influencing a woman's decision to use dietary supplements. AB - Use of dietary supplements by women, particularly those over 40 years of age may be widespread in the United Kingdom. However, from surveillance data, there appears to be a disparity between nutrition and health needs and the rationale for and actual use of dietary supplements by women. This apparent paradox forms the basis for an inverse supplement hypothesis (i.e., supplement use in women appears to be most prevalent among those with least need). Little research has been done to examine the factors underlying the decision to use dietary supplements. Reasons for consuming dietary supplements are often complex, combining social, psychological, knowledge and economic factors. The theory of planned behavior is a widely used model for assessing factors influencing behavioral motivation and action that may be useful for assessing specific diet- and nutrition-related practices. It provided the basis for the development of a questionnaire to explore overall dietary supplement use in a cohort of women in the United Kingdom. The analysis of factors related to beliefs underlying dietary supplement use revealed differences between supplement users and nonusers. Differences included a stronger belief by users than nonusers that taking dietary supplements ensures against possible ill health. Both users and nonusers of supplements also perceived the media (books and magazines) to be a powerful influence on a person's decision to use supplements. These findings highlight the potential of the theory of planned behavior in exploring supplement-taking behavior while throwing light on the factors influencing an individual's motivations to use dietary supplements. PMID- 12771350 TI - Phytoestrogen supplement use by women. AB - Phytoestrogens are weak estrogens found concentrated in soybeans. Americans consume phytoestrogens primarily in traditional soy foods, soymilk and isolated soy protein added during food processing or consumed as a beverage. Extracted phytoestrogens are also marketed in numerous forms as dietary supplements regulated under the Dietary Supplement Health and Education Act. Consumers of phytoestrogen supplements tend to be peri- and postmenopausal women looking for an alternative to hormone therapy. Although there are no approved health claims for phytoestrogens at this time, numerous claims are being made regarding benefits to heart, bone, breast and general menopausal health. The data supporting these claims are generally not strong. The strongest data show that phytoestrogens reduce the number and intensity of hot flashes, although the reduction is a modest 10-20%. The studies showing cholesterol lowering have used soy protein rather than phytoestrogen extracts. The soy protein appears to be required for this effect, although phytoestrogen extracts may have other beneficial effects on the cardiovascular system. The data on bone metabolism are suggestive of possible benefits whereas the effects on the breast are the most poorly understood. Although most animal studies have shown cancer-preventive effects, a few recent studies suggest that soy phytoestrogens may stimulate breast cancer cell growth under certain circumstances. Before recommendations regarding phytoestrogen supplements can be safely made, we must have more information on the effects of the extracts on bone, heart and breast health. Until safety with respect to breast cancer is established, phytoestrogen supplements should not be recommended, particularly for women at high risk of breast cancer. PMID- 12771351 TI - Interaction of science, consumer practices and policy: calcium and bone health as a case study. AB - Data to support a relationship between calcium and bone health are a major part of the body of evidence that underlie calcium-related policy in the United States. Examples of these policies include dietary intake recommendations, health claims for calcium and osteoporosis on food labels and an objective to improve calcium intake of the U.S. population in Healthy People 2010. Median calcium intakes among females fall below recommended levels after childhood even when supplemental calcium intakes are included. This is a concern in light of data that support a positive relationship between calcium and bone health. Most of the studies on the calcium-bone relationship have focused on older women, and several have used fracture as the endpoint; a meta-analysis of their results suggests that increased calcium intake is associated with approximately 30% decrease in fracture risk. Studies in children, adolescents and premenopausal women have focused on the relationship between calcium and bone mineral density rather than fracture; most of these also support a positive relationship between calcium intake and skeletal health although some data gaps remain. Calcium appears to be a threshold nutrient (e.g., intakes above a certain level do not result in further benefit to bone). The effect of increased calcium intake on bone density does not appear to persist unless the higher intakes are sustained. There are certain conditions, such as lactation, during which calcium intake does not appear to influence bone. Other factors that may influence the effect of calcium on bone include bone-specific genotypes and physical activity. PMID- 12771352 TI - Dietary supplement use by American women: challenges in assessing patterns of use, motives and costs. AB - The 1994 passage of Public Law 103-417, the Dietary Supplement and Health Education Act, resulted in an exponential increase in the number and variety of dietary supplements available for over-the-counter purchase. Surveys conducted on random samples of U.S. residents have shown that approximately half of all American women use dietary supplements regularly, but very little is known about the risks and benefits of long-term and widespread supplement use. To accurately evaluate the health effects of supplement use, it is important to characterize usage patterns, motivations and costs of supplement use. However, this is a considerable challenge because accurate supplement data are difficult to collect, product databases with consistent and reliable information are lacking and survey instruments or interview protocols currently in use may not capture information about product selection for specific health conditions, motivations for use, or out-of-pocket expenditures. Future research would benefit from collaborative efforts among governmental scientists, academic scientists and industry to improve dietary supplement data collection methods and develop appropriate tools for analysis. PMID- 12771353 TI - Pregnancy and lactation: physiological adjustments, nutritional requirements and the role of dietary supplements. AB - Nutritional needs are increased during pregnancy and lactation for support of fetal and infant growth and development along with alterations in maternal tissues and metabolism. Total nutrient needs are not necessarily the sum of those accumulated in maternal tissues, products of pregnancy and lactation and those attributable to the maintenance of nonreproducing women. Maternal metabolism is adjusted through the elaboration of hormones that serve as mediators, redirecting nutrients to highly specialized maternal tissues specific to reproduction (i.e., placenta and mammary gland). It is most unlikely that the heightened nutrient needs for successful reproduction can always be met from the maternal diet. Requirements for energy-yielding macronutrients increase modestly compared with several micronutrients that are unevenly distributed among foods. Altered nutrient utilization and mobilization of reserves often offset enhanced needs but sometimes nutrient deficiencies are precipitated by reproduction. There are only limited data from well-controlled intervention studies with dietary supplements and with few exceptions (iron during pregnancy and folate during the periconceptional period), the evidence is not strong that nutrient supplements confer measurable benefit. More research is needed and in future studies attention must be given to subject characteristics that may influence ability to meet maternal and infant demands (genetic and environmental), nutrient-nutrient interactions, sensitivity and selectivity of measured outcomes and proper use of proxy measures. Consideration of these factors in future studies of pregnancy and lactation are necessary to provide an understanding of the links among maternal diet; nutritional supplementation; and fetal, infant and maternal health. PMID- 12771354 TI - National nutrition data: contributions and challenges to monitoring dietary supplement use in women. AB - Survey data from three nationally representative surveys--the National Health and Nutrition Examination Survey, National Health Interview Survey and Continuing Survey of Food Intakes by Individuals--indicate that, in general, women are greater consumers of dietary supplements than men in terms of overall prevalence of use and number of supplements taken. However, monitoring dietary supplement use over time and aggregation or comparison of findings over different surveys is hampered by a lack of comparability between survey data collection and analysis. Differences exist in the types of dietary supplements queried, use of a referent time frame, specificity regarding the supplement taken and level of detail collected relating to personal usage. Some comparability in supplement data collection may be possible but some inconsistencies may persist because of differences in survey goals or collection procedures. Collection of data on dietary supplement use is challenging and collection of very detailed and precise data are time consuming and expensive. Consequently, the level of detail and precision necessary for monitoring, research, and policy uses is an issue that should be addressed in view of the high monetary and time costs of detailed dietary supplement data collection, as well as increased demands on survey respondent time. PMID- 12771355 TI - Dietary supplement use in women: the role of the media. AB - Women and other consumers obtain information on supplements from a variety of sources, including health professionals, but most frequently the media. Although scientific conclusions are methodically scrutinized, news stories are judged by instant appeal. Dietary supplements add a complicating twist because folkloric use predates scientific research by thousands of years. The incongruity between science and the media perpetuates misinformation and fails to provide the context that gives scientific research meaning. The International Food Information Council (IFIC) and the IFIC Foundation(3) seek to bridge the practice and the communication of science. The IFIC Foundation's biannual analysis of food news revealed that in 1999 and 2001, science experts were the primary source of information for articles but context was often absent or incomplete. The media's major obstacle in communicating science is a lack of understanding of the scientific process. It is imperative that emerging science is meaningfully translated for the public. The Harvard School of Public Health and IFIC Foundation convened an advisory group in 1997 to examine the communications process. The result was a set of questions meant to guide the communications process: Will your communication enhance public understanding of diet and health? Have you put the study findings into context? Have the findings been peer reviewed? Have you disclosed the important facts about the study? Have you disclosed all key information about the study's findings? The advisory group agreed that funding sources should be disclosed but that findings ought to stand on their own merit. In addition, guidelines for various specific communicators were created. PMID- 12771356 TI - Dietary supplement use in the context of health disparities: cultural, ethnic and demographic determinants of use. AB - Women of African American, Hispanic, Asian, Pacific Islander, Native American and Alaskan descent constitute 29% of the female population in the United States but they experience health problems disproportionately. Compared with white women as a group, they are in poorer health and use fewer health services. We know from recent studies that the daily use of multivitamins has been associated with lower risk of coronary disease, colon cancer and breast cancer, particularly for alcohol drinkers. In addition, daily multivitamin and multimineral usage by the elderly can reduce the number of days of illness due to infections by 50%. However, supplement use among women tends to be more prevalent among the middle and older age categories; white, well-educated and higher income women; and those residing in the western part of the United States. This examination of the current health disparities and usage patterns indicates that the women who could benefit most from supplements are not typical users. Qualitative data collected on iron and folic acid supplementation programs in developing countries indicate that diverse cultural practices, attitudes and beliefs among vulnerable populations may influence supplement use. However, data in the U.S literature that describe these factors by culture or ethnicity are sparse. If we are to promote dietary supplements to women who are most vulnerable, more research is warranted in the area of health beliefs, attitudes and sociodemographic determinants of supplement use by culture and or ethnicity, particularly among underprivileged groups. PMID- 12771357 TI - Introduction to the 2nd Amino Acid Assessment Workshop. AB - The proportions of amino acids in diets typical of human populations usually differ from the proportions in which they are required, although adverse effects due to such differences are not common. However, there is little systematic information about the adverse effects and the pathophysiological mechanisms of excessive intakes of single or mixtures of amino acids in human subjects. To promote the safe and effective application of amino acids in clinical nutrition and for health promotion it is necessary to establish a sound scientific basis for evaluating their efficacy and safety under various conditions of use. Hence, a series of Amino Acid Assessment Workshops (AAAW) are being organized to bring together experts in amino acid nutrition, metabolism, cell and molecular biology, toxicology and regulation/policy with the eventual purpose of establishing a paradigm for the characterization of risks associated with the ingestion of specific intakes of amino acids by humans. In this introductory paper I summarize the major issues arising at the 1st AAAW, held in Tokyo June, 2001, and provide an introductory context to the present, 2nd AAAW. PMID- 12771358 TI - Application of the concepts of risk assessment to the study of amino acid supplements. AB - Risk assessment is defined as the use of available scientific information to characterize potentially adverse effects associated with exposure of humans to an agent under known or expected conditions. Practically, risk assessment is intended to provide the scientifically sound basis for regulatory or nonregulatory action to manage the risks in humans from the agent. Therefore, the final goal of risk assessment for supplementary use of amino acids is to provide scientific evidence and scientific logic for the establishment of tolerable upper intake levels (UL) for amino acids. At present, however, execution of risk assessment for amino acids is hampered by deficiencies in necessary scientific information, particularly, experimental or clinical/epidemiological data related to the estimation of no-observed-adverse-effect-levels (NOAEL), and scientific principles for the allocation of uncertainty factors (UF) in extrapolation from experimental/clinical data to the general human population. This paper attempts to identify the scientific data and scientific thoughts/methodologies required for deriving UL or assessing the margin of safety for the supplementary use of amino acids. PMID- 12771359 TI - Approaches to risk assessment for macronutrients and amino acids. AB - Risk assessment is a well-established framework for organizing and evaluating diverse, and sometimes conflicting information, to assess the likelihood that agents in the environment may harm human health under known or expected conditions of exposure. Risk assessments are used by regulatory and public health officials to guide judgments and actions regarding the need for risk reduction and the appropriate means to achieve it. These judgments and actions are called risk management, and are guided by law, historical precedent and public health, economic and social concerns. Those in the nutrition community who have been called upon to make recommendations regarding adequate nutrient intakes have long been engaged in the practice of risk assessment. That is, they have assessed the harmful health effects of inadequate intakes and defined intakes likely to avoid such harm. During the past decade attention has turned to the potential health risks of excessive nutrient and nutritional supplement intakes. A recent study released by a committee of the Institute of Medicine illustrates the difficulties in deriving risk-based upper levels of intake for nutrients and nutritional supplements. Amino acids were the subject of extensive discussions by this committee, but in no case was an upper level of intake recommended. It is clear that the extent of scientific investigation of the harmful effects of amino acids has been highly uneven, and that significant questions remain regarding the appropriate methodologies to study such effects. PMID- 12771360 TI - The safety testing of amino acids. AB - The risk assessment of compounds added to foods or taken as supplements is usually based on hazard characterization studies performed in animal test species. A large default uncertainty factor, or margin or exposure, is usually required to allow for possible species differences and human variability in the toxicokinetics and toxicodynamics of the compound. The development of biomarkers offers the potential to rationalize the risk assessment of amino acids, and to refine the extrapolation of data from animals to humans. The use of high resolution nuclear magnetic resonance spectroscopy applied to readily accessible biological fluids, such as urine and plasma, offers great potential for the identification of toxicologically relevant biomarkers in animal studies that can then be applied to studies in humans. PMID- 12771361 TI - Amino acid pharmacokinetics and safety assessment. AB - Tracer kinetic studies of amino acid metabolism during periods of high amino acid intake should allow insights into adaptive or maladaptive regulatory mechanisms controlling amino acid catabolic or disposal events before clinically evident effects. The principles of amino acid tracer kinetics have been well defined, but their application to establishing upper safe intake levels has been essentially nonexistent. Similarly, the pharmacology field has well-established disciplines of toxicokinetics (the relationship of toxicant dose and delivery to its site of action) and toxicodynamics (the relationship of toxicant at its site of action and downstream functional consequences), but these principles have not been transferred to the field of amino acid metabolism. In this context, a theoretical framework is presented for tracer kinetic experiments to help establish upper tolerable levels of amino acid infusion and/or ingestion. In addition, experiments to couple specific amino acid intake levels with their consequent physiological dynamic effects are suggested to lead to the construction of benefit-risk curves that may permit definition of safe amino acid intake ranges for the population. PMID- 12771362 TI - Recent advances in the understanding of amino acid regulation of gene expression. AB - In mammals, the impact of nutrients on gene expression has become an important area of research. Because amino acids have multiple and important functions, their homeostasis has to be finely maintained. However, amino acidemia can be affected by certain nutritional conditions or various forms of stress. Consequently, mammals must adjust several of the physiological functions involved in the adaptation to amino acid availability by regulating expression of numerous genes. It has been shown that amino acids alone can modify the expression of target genes. However, understanding of amino acid-dependent control of gene expression has just started to emerge. This review focuses on recent advances in the understanding of mechanisms involved in the amino acid control of gene expression. PMID- 12771363 TI - Amino acids as regulators of gene expression at the level of mRNA translation. AB - Amino acids act through a number of signaling pathways and mechanisms to mediate control of gene expression at the level of mRNA translation. This report reviews recent findings that illustrate the manner through which amino acids act to regulate the initiation phase of mRNA translation. The report focuses on signaling pathways that involve the eukaryotic initiation factor-2 (eIF2) protein kinase, general control non-derepressing kinase-2 and the mammalian target of rapamycin (mTOR) protein kinase. It also describes the mechanisms through which amino acid-induced modulation of eIF2 phosphorylation and mTOR-mediated signaling cause derepression of translation of specific mRNAs and result in an overall change in the pattern of gene expression. Finally, it provides examples of mRNAs whose translation is modulated through these mechanisms. PMID- 12771364 TI - Amino acids as regulators of proteolysis. AB - Proteolysis, as well as protein synthesis, is a major process that contributes to the body protein turnover. Despite the huge variety of proteases in the body, there are very few proteolytic systems contributing to the complete hydrolysis of proteins to amino acids. The autophagic-lysosomal pathway is responsible for bulk proteolysis, whereas the ubiquitin-proteasome pathway plays a significant role in the fine control of the degradation of specific proteins. Both systems can produce free amino acids as a final product, but only the autophagy system is physiologically controlled by plasma amino acids. Recently, the study of amino acids as regulators of macromolecular turnover has been focused on for their signal transduction mechanism. In autophagic proteolysis, several amino acids have a direct regulatory potential: Leu, Gln, Tyr, Phe, Pro, Met, Trp and His in the liver, and Leu in the skeletal muscle. These amino acids are recognized at the plasma membrane, indicating the possible existence of an amino acid receptor/sensor for their recognition and subsequent intracellular signaling. Another line of evidence has emerged that protein kinase cascades such as mTOR, Erk, eIF2alpha etc. may be involved in the regulation of autophagy, and that amino acids, in combination with insulin, may exert their effects through these pathways. From the viewpoint of amino acid safety, the contribution of proteolysis to possible adverse effects caused by excessive amino acid intake is not clear. At present, there is one report that excess glutamine at 10-fold the plasma level has an abnormal inhibitory effect on hepatic proteolysis, due to a lysosomotropic toxicity of ammonia derived from glutamine degradation. Whether this may lead to an adverse effect in humans remains to be clarified. PMID- 12771365 TI - Amino acids as regulators and components of nonproteinogenic pathways. AB - Amino acids are not only important precursors for the synthesis of proteins and other N-containing compounds, but also participate in the regulation of major metabolic pathways. Glutamate and aspartate, for example, are components of the malate/aspartate shuttle and their concentrations control the rate of mitochondrial oxidation of glycolytic NADH. Glutamate also controls the rate of urea synthesis, not only as the precursor of ammonia and aspartate, but as substrate for synthesis of N-acetylglutamate, the essential activator of carbamoyl-phosphate synthase. This mechanism allows large variations in urea synthesis at relatively constant ammonia concentrations. Increases in intracellular amino acid concentration increase cell volume. Cell swelling per se has anabolic effects on protein, carbohydrate and lipid metabolism: enhanced synthesis of macromolecules compensates for increases in intracellular osmolarity. Mechanisms responsible for cell swelling-induced changes in pathway fluxes include changes in intracellular ion concentrations and in signal transduction. Specific amino acids (e.g., leucine) stimulate protein synthesis and inhibit (autophagic) protein degradation independent of changes in cell volume because they stimulate mTOR (mammalian target of rapamycin), a protein kinase, which is one of the components of a signal transduction pathway used by insulin. When the cellular energy state is low, stimulation of mTOR by amino acids is prevented by activation of AMP-dependent protein kinase. Amino acid dependent signaling also promotes insulin production by beta-cells. This further adds to the anabolic properties of amino acids. It is concluded that amino acids are important regulators of major metabolic pathways. PMID- 12771366 TI - Animal models reveal pathophysiologies of tyrosinemias. AB - The activity of the enzyme 4-hydroxyphenylpyruvic acid dioxygenase (HPD) is regulated by transcription factors. Mutations in the HPD locus are related to two known distinct diseases: hereditary tyrosinemia type 3 and hawkinsinuria. HPD deficient mice are a good model with which to examine the biological effects of 4 hydroxyphenylpyruvic acid, which is a keto acid that causes no apparent visceral damage. In contrast, hereditary tyrosinemia type 1, a genetic disease caused by a deficiency of fumarylacetoacetate hydrolase (FAH), induces severe visceral injuries. Mice with FAH deficiency are lethal after birth; thus, efforts to elucidate the mechanisms of the disease process have been impeded. The use of Fah(-/-) Hpd(-/-) double-mutant mice has enabled studies on tyrosinemias, and essential features of visceral injury have been reveale. PMID- 12771367 TI - Interorgan amino acid transport and its regulation. AB - Interorgan amino acid transport is a highly active and regulated process that provides amino acids to all tissues of the body, both for protein synthesis and to enable amino acids to be used for specific metabolic functions. It is also an important component of plasma amino acid homeostasis. Net movement of amino acids depends on the physiological and nutritional state. For example, in the fed state the dominant flux is from the intestine to the other tissues. In starvation the dominant flux is from muscle to the liver and kidney. A number of general principles underlie many amino acid fluxes: i) The body does not have a store for amino acids. This means that dietary amino acids, in excess of those required for protein synthesis, are rapidly catabolized; ii) Amino acid catabolism must occur in a manner that does not elevate blood ammonia. Thus, extrasplanchnic amino acid metabolism often involves an innocuous means of transporting nitrogen to the liver; iii) Because most amino acids are glucogenic, there will be a considerable flux of amino acids to the gluconeogenic organs when there is a need to produce glucose. In addition to these bulk flows, fluxes of many specific amino acids underlie specific organ function. These include intestinal oxidation of enteral amino acids, the intestinal/renal axis for arginine production, the brain uptake of neurotransmitter precursors and renal glutamine metabolism. There is no single means of regulating amino acid fluxes; rather, such varied mechanisms as substrate supply, enzyme activity, transporter activity and competitive inhibition of transport are all found. PMID- 12771368 TI - Evaluation of the effects of the dietary intake of proteins and amino acids by DNA microarray technology. AB - The DNA microarray technique has been increasingly utilized in various fields of life sciences. It allows us to analyze the expression levels of thousands of genes simultaneously. The high productivity will facilitate the evaluation of changes in amino acid metabolism and their consequences in response to dietary proteins and amino acids. We compared the expression profiles by the GeneChip system in the liver and other tissues among three groups of rats fed with a 12% casein, a 12% gluten or a protein-free diet. Feeding the gluten or the protein free diet up- or down-regulated a few hundred genes in the liver compared to the casein diet. Although some of the genes were already known to respond to changes in the protein nutritional state, the majority was newly identified responders. This paper also discusses the possibility of a use this technology for safety evaluation of excessive intake of dietary components, especially of amino acids. PMID- 12771369 TI - Genomics and metabolomics as markers for the interaction of diet and health: lessons from lipids. AB - Foods are not purified compounds acting on single molecular targets, but complex mixtures of molecules that modulate many biochemical pathways simultaneously. Diet affects the probability of developing various diseases. Nevertheless, specific recommendations for individual diets are not simple. Recommending nutrient intakes above and beyond those needed to provide adequacy requires scientific knowledge and regulatory scrutiny to ensure the efficacy and safety even of essential nutrients. Designing a diet to improve metabolic health is a bold and ambitious goal. It is possible to design foods that will alter metabolism, but what change will make everyone who is otherwise healthy even healthier? Changing one aspect of metabolism to lower the risk of one disease does not improve overall health if it comes at the expense of disrupting another aspect of metabolism that increases the risk of another disease. This issue has: 1) frustrated nutritional recommendations that could provide benefits to the health of large subsets of the population, 2) caused the recall of drugs with many beneficial effects and 3) caused harm by implying that single nutrients/foods could be healthy for everyone. An individualized system for metabolic assessment would establish the efficacy and safety of nutrients such as amino acids or fatty acids when these are designed to be consumed at levels providing improved metabolic health. The need to document the lack of an adverse effect of a food or drug on physiology necessitates a global, i.e. metabolomic approach. PMID- 12771370 TI - Plasma protein synthesis measurements using a proteomics strategy. AB - The analysis of the synthesis of proteins has been the subject of many studies in animals and humans. Plasma proteins can be used as an easy accessible source of specific proteins. In this paper, an innovative method to study the synthetic rate of plasma proteins is described. This methodology, based on the proteomics approach, enables the direct observation of the effects of posttranslational modifications of protein synthesis and/or degradation. The methodology is based on 1D or 2D electrophoresis and subsequent electrospray ionization liquid chromatography mass spectrometry (ESI-LC-MS). Protein synthesis is measured in isotopically labeled peptides of the identified proteins. This innovative method can be used to assess amino acid adequacy and safety by studying protein synthesis and posttranslational modification of plasma proteins in more detail. PMID- 12771371 TI - Proteomics: advanced technology for the analysis of cellular function. AB - Proteomics developed initially from the decade-long study of comprehensive protein visualization on two-dimensional electrophoresis gels has been expanded by mass spectrometry and the growth in searchable sequence databases. Currently, by use of more sophisticated technology such as a combination of multidimensional chromatography and mass spectrometry, thousands of proteins can automatically be identified in a day along with semiquantitative information on differential protein expression. As with differential gene expression by cDNA-chips, the differential-protein analysis is useful for monitoring and identifying proteins involved in various physiological changes in cells or organisms, although the analysis alone does not necessarily provide information regarding the cause of the change or the function of the proteins. However, proteomics also provides the tools to expand into more sophisticated biochemical approaches, such as the study of protein interactions that can be determined directly by performing a pull-down assay with a bait protein followed by mass spectrometric identification of the bound proteins. Proteomics, thus, is useful for both large-scale surveys of proteins and detailed studies of the functional relationships among the proteins of interest. Certainly this approach can be applicable to the assessment of amino acid adequacy and safety. PMID- 12771373 TI - General discussion at 2nd Amino Acid Assessment Workshop (2nd AAAW). PMID- 12771372 TI - Metabolomics and its potential for assessment of adequacy and safety of amino acid intake. AB - In this short paper, we suggest some concepts and methods that may be useful in considering a metabolomic approach to the study of amino acid adequacy and safety. We suggest that the analysis of a metabolomic subset, such as amino acids, may yield useful information, and that correlation-based analyses could be useful in the analysis of metabolomic data to determine which metabolites may be responsible for the effects of excessive intakes of amino acids. The use of correlation-based analyses would allow the use of arbitrarily scaled pseudoquantitative data, making it possible to analyze unidentified peaks, which may be observed when the object sample has gone through chromatographic analysis, together with known ones. We suggest that these basic concepts and methods could serve as the basis for a metabolomic approach for the assessment of the range of adequacy of amino acid intakes. PMID- 12771374 TI - Folding and misfolding of the papillomavirus E6 interacting peptide E6ap. AB - All-atom Langevin dynamics simulations have been performed to study the folding pathways of the 18-residue binding domain fragment E6ap of the human papillomavirus E6 interacting peptide. Six independent folding trajectories, with a total duration of nearly 2 micros, all lead to the same native state in which the E6ap adopts a fluctuating alpha-helix structure in the central portion (Ser-4 Leu-13) but with very flexible N and C termini. Simulations starting from different core configurations exhibit the E6ap folding dynamics as either a two- or three-state folder with an intermediate misfolded state. The essential leucine hydrophobic core (Leu-9, Leu-12, and Leu-13) is well conserved in the native state structure but absent in the intermediate structure, suggesting that the leucine core is not only essential for the binding activity of E6ap but also important for the stability of the native structure. The free energy landscape reveals a significant barrier between the basins separating the native and misfolded states. We also discuss the various underlying forces that drive the peptide into its native state. PMID- 12771375 TI - Peripheral heme substituents control the hydrogen-atom abstraction chemistry in cytochromes P450. AB - We elucidate the hydroxylation of camphor by cytochrome P450 with the use of density functional and mixed quantum mechanics/molecular mechanics methods. Our results reveal that the enzyme catalyzes the hydrogen-atom abstraction step with a remarkably low free-energy barrier. This result provides a satisfactory explanation for the experimental failure to trap the proposed catalytically competent high-valent heme Fe(IV) oxo (oxyferryl) species responsible for this hydroxylation chemistry. The primary and previously unappreciated contribution to stabilization of the transition state is the interaction of positively charged residues in the active-site cavity with carboxylate groups on the heme periphery. A similar stabilization found in dioxygen binding to hemerythrin, albeit with reversed polarity, suggests that this mechanism for controlling the relative energetics of redox-active intermediates and transition states in metalloproteins may be widespread in nature. PMID- 12771376 TI - Crystal structure of soybean 11S globulin: glycinin A3B4 homohexamer. AB - Most plant seeds contain 11S globulins as major storage proteins for their nutrition. Soybean glycinin belongs to the 11S globulin family and consists of five kinds of subunits. We determined the crystal structure of a homohexamer of the glycinin A3B4 subunit at 2.1-A resolution. The crystal structure shows that the hexamer has 32-point group symmetry formed by face-to-face stacking of two trimers. The interface buries the highly conserved interchain disulfide. Based on the structure, we propose that an ingenious face-to-face mechanism controls the hexamer formation of the 11S globulin by movement of a mobile disordered region to the side of the trimer after posttranslational processing. Electrostatic analysis of the faces suggests that the interchain disulfide-containing face has high positive potential at acidic pH, which induces dissociation of the hexamer into trimers that may be susceptible to proteinases after seed imbibition. This dissociation might result in the degradation and mobilization of 11S globulins as storage proteins in embryos during germination and seedling growth. PMID- 12771377 TI - Spatial parasite transmission, drug resistance, and the spread of rare genes. AB - The transmission of many parasitic worms involves aggregated movement between hosts of "packets" of infectious larvae. We use a generic metapopulation model to show that this aggregation naturally promotes the preferential spread of rare recessive genes, compared with the expectations of traditional nonspatial models. A more biologically realistic model also demonstrates that this effect could explain the rapid observed spread of recessive or weakly dominant drug-resistant genotypes in nematode parasites of sheep. This promotion of a recessive trait arises from a novel mechanism of inbreeding arising from the metapopulation dynamics of transmission. PMID- 12771378 TI - Absolute quantification of proteins and phosphoproteins from cell lysates by tandem MS. AB - A need exists for technologies that permit the direct quantification of differences in protein and posttranslationally modified protein expression levels. Here we present a strategy for the absolute quantification (termed AQUA) of proteins and their modification states. Peptides are synthesized with incorporated stable isotopes as ideal internal standards to mimic native peptides formed by proteolysis. These synthetic peptides can also be prepared with covalent modifications (e.g., phosphorylation, methylation, acetylation, etc.) that are chemically identical to naturally occurring posttranslational modifications. Such AQUA internal standard peptides are then used to precisely and quantitatively measure the absolute levels of proteins and posttranslationally modified proteins after proteolysis by using a selected reaction monitoring analysis in a tandem mass spectrometer. In the present work, the AQUA strategy was used to (i) quantify low abundance yeast proteins involved in gene silencing, (ii) quantitatively determine the cell cycle-dependent phosphorylation of Ser-1126 of human separase protein, and (iii) identify kinases capable of phosphorylating Ser-1501 of separase in an in vitro kinase assay. The methods described here represent focused, alternative approaches for studying the dynamically changing proteome. PMID- 12771379 TI - The BglF sensor recruits the BglG transcription regulator to the membrane and releases it on stimulation. AB - The Escherichia coli BglF protein is a sugar-sensor that controls the activity of the transcriptional antiterminator BglG by reversibly phosphorylating it, depending on beta-glucoside availability. BglF is a membrane-bound protein, whereas BglG is a soluble protein, and they are both present in the cell in minute amounts. How do BglF and BglG find each other to initiate signal transduction efficiently? Using bacterial two-hybrid systems and the Far-Western technique, we demonstrated unequivocally that BglG binds to BglF and to its active site-containing domain in vivo and in vitro. Measurements by surface plasmon resonance corroborated that the affinity between these proteins is high enough to enable their stable binding. To visualize the subcellular localization of BglG, we used fluorescence microscopy. In cells lacking BglF, the BglG-GFP fusion protein was evenly distributed throughout the cytoplasm. In contrast, in cells producing BglF, BglG-GFP was localized to the membrane. On addition of beta glucoside, BglG-GFP was released from the membrane, becoming evenly distributed throughout the cell. Using mutant proteins and genetic backgrounds that impede phosphorylation of the Bgl proteins, we demonstrated that BglG-BglF binding and recruitment of BglG to the membrane sensor requires phosphorylation but does not depend on the individual phosphorylation sites of the Bgl proteins. We suggest a mechanism for rapid response to environmental changes by preassembly of signaling complexes, which contain transcription regulators recruited by their cognate sensors-kinases, under nonstimulating conditions, and release of the regulators to the cytoplasm on stimulation. This mechanism might be applicable to signaling cascades in prokaryotes and eukaryotes. PMID- 12771380 TI - Apparent homology of expressed genes from wood-forming tissues of loblolly pine (Pinus taeda L.) with Arabidopsis thaliana. AB - Pinus taeda L. (loblolly pine) and Arabidopsis thaliana differ greatly in form, ecological niche, evolutionary history, and genome size. Arabidopsis is a small, herbaceous, annual dicotyledon, whereas pines are large, long-lived, coniferous forest trees. Such diverse plants might be expected to differ in a large number of functional genes. We have obtained and analyzed 59,797 expressed sequence tags (ESTs) from wood-forming tissues of loblolly pine and compared them to the gene sequences inferred from the complete sequence of the Arabidopsis genome. Approximately 50% of pine ESTs have no apparent homologs in Arabidopsis or any other angiosperm in public databases. When evaluated by using contigs containing long, high-quality sequences, we find a higher level of apparent homology between the inferred genes of these two species. For those contigs 1,100 bp or longer, approximately 90% have an apparent Arabidopsis homolog (E value < 10-10). Pines and Arabidopsis last shared a common ancestor approximately 300 million years ago. Few genes would be expected to retain high sequence similarity for this time if they did not have essential functions. These observations suggest substantial conservation of gene sequence in seed plants. PMID- 12771381 TI - Glycerol replacement corrects defective skin hydration, elasticity, and barrier function in aquaporin-3-deficient mice. AB - Mice deficient in the epidermal water/glycerol transporter aquaporin-3 (AQP3) have reduced stratum corneum (SC) hydration and skin elasticity, and impaired barrier recovery after SC removal. SC glycerol content is reduced 3-fold in AQP3 null mice, whereas SC structure, protein/lipid composition, and ion/osmolyte content are not changed. We show here that glycerol replacement corrects each of the defects in AQP3 null mice. SC water content, measured by skin conductance and 3H2O accumulation, was 3-fold lower in AQP3 null vs. wild-type mice, but became similar after topical or systemic administration of glycerol in quantities that normalized SC glycerol content. SC water content was not corrected by glycerol like osmolytes such as xylitol, erythritol, and propanediol. Orally administered glycerol fully corrected the reduced skin elasticity in AQP3 null mice as measured by the kinetics of skin displacement after suction, and the delayed barrier recovery as measured by transepidermal water loss after tape-stripping. Analysis of [14C]glycerol kinetics indicated reduced blood-to-SC transport of glycerol in AQP3 null mice, resulting in slowed lipid biosynthesis. These data provide functional evidence for a physiological role of glycerol transport by an aquaglyceroporin, and indicate that glycerol is a major determinant of SC water retention, and mechanical and biosynthetic functions. Our findings establish a scientific basis for the >200-yr-old empirical practice of including glycerol in cosmetic and medicinal skin formulations. PMID- 12771382 TI - The transient receptor potential channel on the yeast vacuole is mechanosensitive. AB - Ca2+ is released from the vacuole into the yeast cytoplasm on an osmotic upshock, but how this upshock is perceived was unknown. We found the vacuolar channel, Yvc1p, to be mechanosensitive, showing that the Ca2+ conduit is also the sensing molecule. Although fragile, the yeast vacuole allows limited direct mechanical examination. Pressures at tens of millimeters of Hg (1 mmHg = 133 Pa) activate the 400-pS Yvc1p conductance in whole-vacuole recording mode as well as in the excised cytoplasmic-side-out mode. Raising the bath osmolarity activates this channel and causes vacuolar shrinkage and deformation. It appears that, on upshock, a transient osmotic force activates Yvc1p to release Ca2+ from the vacuole. Mechanical activation of Yvc1p occurs regardless of Ca2+ concentration and is apparently independent of its known Ca2+ activation, which we now propose to be an amplification mechanism (Ca2+-induced Ca2+ release). Yvc1p is a member of the transient receptor potential-family channels, several of which have been associated with mechanosensation in animals. The possible use of Yvc1p as a molecular model to study mechanosensation in general is discussed. PMID- 12771383 TI - Effective behavioral treatment of focal hand dystonia in musicians alters somatosensory cortical organization. AB - New perspectives in neurorehabilitation suggest that behavioral treatments of movement disorders may modify the functional organization of central somatosensory neural networks. On the basis of the assumption that use-dependent reorganization in these networks contributes to the fundamental abnormalities seen in focal dystonia, we treated 10 affected musicians and measured the concomitant somatosensory changes by using whole-head magnetoencephalography. We found that effective treatment, using the method of sensory motor retuning, leads to alterations in the functional organization of the somatosensory cortex. Specifically, before treatment, somatosensory relationships of the individual fingers differ between the affected and unaffected hands, whereas after treatment, finger representations contralateral to the dystonic side become more similar to the less-affected side. Further, somatosensory finger representations are ordered more according to homuncular principles after treatment. In addition, the observed physiologic changes correlated with behavioral data. These results confirm that plastic changes in parallel with emergent neurological dysfunction may be reversed by context-specific, intensive training-based remediation. PMID- 12771385 TI - Recombineering with overlapping single-stranded DNA oligonucleotides: testing a recombination intermediate. AB - A phage lambda-based recombination system, Red, can be used for high-efficiency mutagenesis, repair, and engineering of chromosomal or episomal DNA in vivo in Escherichia coli. When long linear double-stranded DNA with short flanking homologies to their targets are used for the recombination, the lambda Exo, Beta, and Gam proteins are required. The current model is: (i) Gam inhibits the host RecBCD activity, thereby protecting the DNA substrate for recombination; (ii) Exo degrades from each DNA end in a 5' --> 3' direction, creating double-stranded DNA with 3' single-stranded DNA tails; and (iii) Beta binds these 3' overhangs to protect and anneal them to complementary sequences. We have tested this model for Red recombination by using electroporation to introduce overlapping, complementary oligonucleotides that when annealed in vivo approximate the recombination intermediate that Exo should create. Using this technique we found Exo-independent recombination. Surprisingly, a similarly constructed substrate with 5' overhangs recombined more efficiently. This 5' overhang recombination required both Exo and Beta for high levels of recombination and the two oligonucleotides need to overlap by only 6 bp on their 3' ends. Results indicate that Exo may load Beta onto the 3' overhang it produces. In addition, multiple overlapping oligonucleotides were successfully used to generate recombinants in vivo, a technique that could prove useful for many genetic engineering procedures. PMID- 12771387 TI - Protection of lungs from hyperoxic injury: gene expression analysis of cyclosporin A therapy. AB - We have previously shown that cyclosporin A (CsA), an inhibitor of protein phosphatase 2B (calcineurin), attenuates hyperoxia-induced reductions in murine lung compliance. CsA protected against hyperoxia-induced changes in neutrophil infiltration, capillary congestion, edema, and hyaline membrane formation. Gene expression studies were conducted to identify the gene expression patterns underlying the protective effects of CsA during hyperoxic lung injury. After 72 h of simultaneous treatment with >95% oxygen and CsA (50 mg x kg(-1) x day(-1)), RNA was isolated from murine lungs. RNA from treated and untreated lungs was reverse transcribed to cDNA, competitively hybridized, and used to probe 8,734 complimentary DNAs on the Incyte mouse GEM 1 array. Several known genes and expressed sequence tags (ESTs) showed increased (GenBank accession numbers: AA125385, AA241295, W87197, syntaxin, and cyclin G) or decreased [AA036517, AA267567, AA217009, W82577, uteroglobin, stromal cell-derived factor 1, and surfactant protein C (SP-C)] expression after hyperoxia. Hyperoxia-stimulated reductions in SP-C gene expression were confirmed through Northern blot analysis. The increase in gene expression of one expressed sequence tag (AA125385) with hyperoxia was reversed by CsA treatment. Sequence data demonstrated that this EST has high homology to murine cyclin B1. Western blot analysis did not demonstrate any changes in distal lung cyclin B1 expression after hyperoxia. Protein expression of cyclin B1 in the distal lung was observed in the endothelial cells, bronchiolar epithelial cells, and both the type I and type II alveolar epithelial cells. Further analysis of cyclin B1 may elucidate the protective actions of CsA in hyperoxic injury. PMID- 12771384 TI - RGS3 mediates a calcium-dependent termination of G protein signaling in sensory neurons. AB - G proteins modulate synaptic transmission. Regulators of G protein signaling (RGS) proteins accelerate the intrinsic GTPase activity of Galpha subunits, and thus terminate G protein activation. Whether RGS proteins themselves are under cellular control is not well defined, particularly in native cells. In dorsal root ganglion neurons overexpressing RGS3, we find that G protein signaling is rapidly terminated (or "desensitized") by calcium influx through voltage-gated channels. This rapid desensitization is most likely mediated by direct binding of calcium to RGS3, as deletion of an EF-hand domain in RGS3 abolishes both the desensitization (observed physiologically) and a calcium-RGS3 interaction (observed in a gel-shift assay). A naturally occurring variant of RGS3 that lacks the EF hand neither binds calcium nor produces rapid desensitization, giving rise instead to a slower calcium-dependent desensitization that is attenuated by a calmodulin antagonist. Thus, activity-evoked calcium entry in sensory neurons may provide differential control of G protein signaling, depending on the isoform of RGS3 expressed in the cells. In complex neural circuits subjected to abundant synaptic inhibition by G proteins (as occurs in dorsal spinal cord), rapid termination of inhibition by electrical activity by EF hand-containing RGS3 may ensure the faithful transmission of information from the most active sensory inputs. PMID- 12771388 TI - The economic burden of pneumoconiosis in China. PMID- 12771386 TI - Probucol prevents early coronary heart disease and death in the high-density lipoprotein receptor SR-BI/apolipoprotein E double knockout mouse. AB - Mice with homozygous null mutations in the high-density lipoprotein receptor SR BI (scavenger receptor class B, type I) and apolipoprotein E genes fed a low-fat diet exhibit a constellation of pathologies shared with human atherosclerotic coronary heart disease (CHD): hypercholesterolemia, occlusive coronary atherosclerosis, myocardial infarctions, cardiac dysfunction (heart enlargement, reduced systolic function and ejection fraction, and ECG abnormalities), and premature death (mean age 6 weeks). They also exhibit a block in RBC maturation and abnormally high plasma unesterified-to-total cholesterol ratio (0.8) with associated abnormal lipoprotein morphology (lamellar/vesicular and stacked discoidal particles reminiscent of those in lecithin/cholesterol acyltransferase deficiency and cholestasis). Treatment with the lipid-lowering, antiatherosclerosis, and antioxidation drug probucol extended life to as long as 60 weeks (mean 36 weeks), and at 5-6 weeks of age, virtually completely reversed the cardiac and most RBC pathologies and corrected the unesterified to total cholesterol ratio (0.3) and associated distinctive abnormal lipoprotein morphologies. Manipulation of the timing of administration and withdrawal of probucol could control the onset of death and suggested that critical pathological changes usually occurred in untreated double knockout mice between approximately 3 (weaning) and 5 weeks of age and that probucol delayed heart failure even after development of substantial CHD. The ability of probucol treatment to modulate pathophysiology in the double knockout mice enhances the potential of this murine system for analysis of the pathophysiology of CHD and preclinical testing of new approaches for the prevention and treatment of cardiovascular disease. PMID- 12771389 TI - Lung hyperpermeability and asthma prevalence in schoolchildren: unexpected associations with the attendance at indoor chlorinated swimming pools. AB - AIMS: To study whether exposure to nitrogen trichloride in indoor chlorinated pools may affect the respiratory epithelium of children and increase the risk of some lung diseases such as asthma. METHODS: In 226 healthy children, serum surfactant associated proteins A and B (SP-A and SP-B), 16 kDa Clara cell protein (CC16), and IgE were measured. Lung specific proteins were measured in the serum of 16 children and 13 adults before and after exposure to NCl(3) in an indoor chlorinated pool. Relations between pool attendance and asthma prevalence were studied in 1881 children. Asthma was screened with the exercise induced bronchoconstriction test (EIB). RESULTS: Pool attendance was the most consistent predictor of lung epithelium permeability. A positive dose-effect relation was found with cumulated pool attendance and serum SP-A and SP-B. Serum IgE was unrelated to pool attendance, but correlated positively with lung hyperpermeability as assessed by serum SP-B. Changes in serum levels of lung proteins were reproduced in children and adults attending an indoor pool. Serum SP-A and SP-B were already significantly increased after one hour on the pool side without swimming. Positive EIB and total asthma prevalence were significantly correlated with cumulated pool attendance indices. CONCLUSIONS: Regular attendance at chlorinated pools by young children is associated with an exposure dependent increase in lung epithelium permeability and increase in the risk of developing asthma, especially in association with other risk factors. We therefore postulate that the increasing exposure of children to chlorination products in indoor pools might be an important cause of the rising incidence of childhood asthma and allergic diseases in industrialised countries. Further epidemiological studies should be undertaken to test this hypothesis. PMID- 12771390 TI - Clinical and histopathological changes of the nasal mucosa induced by occupational exposure to sulphuric acid mists. AB - AIMS: To assess potential alterations of the nasal mucosa by clinical and histopathological evaluation of workers exposed to sulphuric acid mists at anodising plants, correlating the findings with duration of exposure and sulphuric acid concentrations in the air, and comparing them with a control group. METHODS: Fifty two workers from five plants underwent a clinical evaluation (standard questionnaire, clinical, and ear, nose, and throat examination including nasal endoscopy). For the histopathological study, 20 of the 52 subjects (study group) were randomly selected, as well as 11 unexposed subjects (control group), matched by sex, age, and smoking habits. Nasal biopsy specimens were obtained from the anterior septum mucosa and the anterior curvature of the middle turbinate in each individual. A total of 56 nasal mucosa specimens (37 in the study group and 19 in the control group) were evaluated with regard to normal respiratory epithelium or metaplastic epithelium, atypia or dysplasia, and alterations of the lamina propria. RESULTS: The histopathological study revealed squamous metaplasia in 29 (79%) and atypia in 13 (35%) of the 37 study group samples. No association was found between exposure duration and the clinical and histopathological variables, but a significant association was found between sulphuric acid concentrations higher than 200 micro g/m(3) and pale mucosal patches and ulcerations in the exposed subjects. Logistic regression analysis showed that the exposed subjects had a fivefold risk of developing atypia compared with the unexposed subjects. CONCLUSIONS: Workers exposed to sulphuric acid mists presented with a high incidence of nasal symptoms, and macroscopic and microscopic changes of the nasal mucosa, including squamous atypia and dysplasia. The risk for these histopthological lesions increased with higher sulphuric acid concentrations in the air, revealing an exposure-response relation. PMID- 12771392 TI - Radiographic abnormalities among construction workers exposed to quartz containing dust. AB - BACKGROUND: Construction workers are exposed to quartz containing respirable dust, at levels that may cause fibrosis in the lungs. Studies so far have not established a dose-response relation for radiographic abnormalities for this occupational group. AIMS: To measure the extent of radiographic abnormalities among construction workers primarily exposed to quartz containing respirable dust. METHODS: A cross sectional study on radiographic abnormalities indicative of pneumoconiosis was conducted among 1339 construction workers mainly involved in grinding, (jack)-hammering, drilling, cutting, sawing, and polishing. Radiological abnormalities were determined by median results of the 1980 International Labour Organisation system of three certified "B" readers. Questionnaires were used for assessment of occupational history, presence of respiratory diseases, and symptoms and smoking habits. RESULTS: An abnormality of ILO profusion category 1/0 and greater was observed on 10.2% of the chest radiographs, and profusion category of 1/1 or greater on 2.9% of the radiographs. The average duration of exposure of this group was 19 years and the average age was 42. The predominant type of small opacities (irregularly shaped) is presumably indicative of mixed dust pneumoconiosis. The prevalence of early signs of nodular silicosis (small rounded opacities of category 1/0 or greater) was low (0.8%). CONCLUSIONS: The study suggests an elevated risk of radiographic abnormalities among these workers with expected high exposure. An association between radiographic abnormalities and cumulative exposure to quartz containing dust from construction sites was observed, after correction for potentially confounding variables. PMID- 12771391 TI - DNA-protein crosslinks and p53 protein expression in relation to occupational exposure to formaldehyde. AB - BACKGROUND: Formaldehyde (FA) is classified as a probable human carcinogen. AIMS: To examine DNA protein crosslinks (DPC) and p53, which are generally known to be involved in carcinogenesis, in peripheral blood lymphocytes of workers exposed to FA. METHODS: DPC and p53 ("wild type" and mutant) were examined in peripheral blood lymphocytes of 186 workers exposed to FA (mean years of exposure = 16) and 213 unexposed workers. Every worker completed a questionnaire on demographic data, occupational and medical history, smoking, and hygiene. RESULTS: The adjusted mean level of DPC in the exposed and the unexposed workers differed significantly. Adjustment was made for age, sex, years of education, smoking, and origin. Exposure to FA increased the risk of having a higher level of pantropic p53 above 150 pg/ml (OR 1.6, 95% CI 0.8 to 3.1). A significant positive correlation was found between the increase of pantropic p53 protein and mutant p53 protein, as well as between pantropic p53 >150 pg/ml and mutant p53 protein. In the exposed group a significantly higher proportion of p53 >150 pg/ml was found among workers with DPC >0.187 (55.7%) (0.187 = median level of DPC) than among workers with DPC < or =0.187 (33.3%). The risk of having pantropic p53 protein >150 pg/ml was determined mainly by levels of DPC. Workers with DPC above the median level had a significantly higher risk of having pantropic p53 >150 pg/ml (adjusted OR 2.5, 95% CI 1.2 to 5.4). CONCLUSIONS: Results suggest that DPC and mutation in p53 may represent steps in FA carcinogenesis and a possible causal relation between DPC and mutation in p53. These biomarkers can be applied in the assessment of the development of cancer due to FA exposure. PMID- 12771393 TI - Evaluation of decompression safety in an occupational diving group using self reported diving exposure and health status. AB - BACKGROUND: Many occupational diving groups have substantially different diving patterns to those for which decompression schedules are validated. AIMS: To evaluate tuna farm occupational diving practice against existing decompression models and describe a method for collecting and modelling self reported field decompression data. METHODS: Machine readable objective depth/time profiles were obtained from depth/time recorders worn by tuna farm occupational divers. Divers' health status was measured at the end of each working day using a self administered health survey that produces an interval diver health score (DHS) with possible values ranging from 0 to 30. Depth/time profiles were analysed according to existing decompression models. The contribution of diving exposure and between diver variability to DHS was evaluated using linear regression. RESULTS: The mean risk of decompression sickness was calculated as 0.005 (SD 0.003, n = 383). The mean DHS following diving was 3 (SD 2, n = 383) and following non-diving activities was 1 (SD 1, n = 41). After accounting for between diver variability in intercept, DHS was found to increase one unit for every 1% increase in the risk of decompression sickness. CONCLUSIONS: A method has been established for the collection and analysis of self reported objective decompression data from occupational diving groups that can potentially be used as the basis for development of purpose designed occupational diving decompression schedules. PMID- 12771394 TI - Lung cancer risk in workers exposed to poly(vinyl chloride) dust: a nested case referent study. AB - BACKGROUND: There have been few investigations of an association between poly(vinyl chloride) (PVC) dust exposure and an increase in lung cancer incidence, and their conclusions have been inconsistent. AIMS: To determine whether PVC and/or vinyl chloride monomer (VCM) is the associated risk factor(s), by means of a nested case-referent study, in order to estimate lung cancer risk, avoiding selection, information, or confounding biases. METHODS: Thirty eight cases of histologically verified lung cancer and 224 control subjects without a history of cancer were selected from an Italian cohort of 1658 vinyl chloride workers. Information sources included clinical records (diagnosis, smoking habits) and plant records (occupational history). The risk of lung cancer was estimated by odds ratios (OR) with 95% confidence intervals (CI), calculated using logistic regression models. RESULTS: In PVC baggers exposed to high levels of respirable PVC particles in the workplace, the lung cancer OR increases by 20% for each extra year of work (OR = 1.2003; 95% CI 1.0772 to 1.3469; p = 0.0010), when the influence of age and smoking habits is controlled. No relation was found between lung cancer and cumulative VCM exposure. CONCLUSION: This nested case control study showed, in the VCM/PVC industry, an increased risk of lung cancer associated with exposure to PVC dust; previous cohort studies failed to recognise such excess, probably because they used VCM exposure as the risk indicator. PMID- 12771395 TI - Reducing long term sickness absence by an activating intervention in adjustment disorders: a cluster randomised controlled design. AB - AIMS: To compare an innovative activating intervention with "care as usual" (control group) for the guidance of employees on sickness leave because of an adjustment disorder. It was hypothesised that the intervention would be more effective than care as usual in lowering the intensity of symptoms, increasing psychological resources, and decreasing sickness leave duration. METHODS: A prospective, cluster randomised controlled trial was carried out with 192 patients on first sickness leave for an adjustment disorder. Symptom intensity, sickness duration, and return to work rates were measured at 3 months and 12 months. Analyses were performed on an intention to treat basis. RESULTS: At 3 months, significantly more patients in the intervention group had returned to work compared with the control group. At 12 months all patients had returned to work, but sickness leave was shorter in the intervention group than in the control group. The recurrence rate was lower in the intervention group. There were no differences between the two study groups with regard to the decrease of symptoms. At baseline, symptom intensity was higher in the patients than in a normal reference population, but decreased over time in a similar manner in both groups to approximately normal levels. CONCLUSION: The experimental intervention for adjustment disorders was successful in shortening sick leave duration, mainly by decreasing long term absenteeism. PMID- 12771396 TI - Evaluation of the quality of coding of job episodes collected by self questionnaires among French retired men for use in a job-exposure matrix. AB - BACKGROUND: and AIMS: The ESPACES study was intended to identify retirees who may have been, according to their job descriptions, exposed to asbestos during their working lives. As part of this study, we analysed the quality of the occupation and activity sector coding as well as its effect on the subjects' exposure status. METHODS: The occupation and activity sector for a sample of 450 retired men were coded twice (with the second coder blinded to the first result), according to the international codes for industries (ISIC-1975) and occupations (ISCO-1968). For each series, linking the information about a job episode (dates, ISIC code, ISCO code) with the matrix allowed attribution of a probability of asbestos exposure to each episode and each subject. The asbestos exposure in the two series was compared by the kappa reproducibility coefficient. RESULTS: The analysis concerned 425 questionnaires. There was at least one difference in the code for either activity sector (ISIC) or occupation (ISCO) in half the episodes (50.2%). The exposure status estimated by the job-exposure matrix did not change between the series for 84.7% of the subjects. The kappa coefficient was 0.64 for all questionnaires, 0.70 when the questionnaire was coded twice by the same coder, and 0.62 when coded by two different coders. CONCLUSIONS: Despite intra- and inter-differences between coders, the coding of job episodes for the ESPACES study appears satisfactory and hence indicates that the assessment of the subjects' asbestos exposure was assessed without major distortions. This study underlines the usefulness of employing coders specifically trained for this technique. PMID- 12771397 TI - Upper airway inflammation in waste handlers exposed to bioaerosols. AB - AIMS: To examine work associated upper airway inflammation in 31 waste handlers, and to correlate these findings with personally monitored exposure to different bioaerosol components. METHODS: Cell differentials, interleukin 8 (IL-8), myeloperoxidase (MPO), and eosinophilic cationic protein (ECP) were examined in NAL (nasal lavage), and swelling of the nasal mucosa was determined by acoustic rhinometry before work start on Monday and the following Thursday. Bioaerosol exposure was determined by personal full shift exposure measurements on Monday, Tuesday, and Wednesday and analysed for total bacteria, fungal spores, endotoxin, and beta(1-->3)-glucans. RESULTS: The increased percentage of neutrophils from Monday (28%) to Thursday (46%) correlated with increases in ECP (r(S) = 0.71, p < 0.001) and MPO (r(S) = 0.38, p < 0.05), and showed a close to significant correlation with nasal swelling (r(S) = -0.55, p = 0.07). The Thursday levels of neutrophils, MPO, and IL-8 were associated with the exposure to fungal spores (range 0-2.0 x 10(6)/m(3)) and endotoxin (range 4-183 EU/m(3)) measured the day before, and the median exposure to beta(1-->3)-glucans (range 3-217 ng/m(3)), respectively (r(S) = 0.47-0.54, p < 0.01). Swelling of the nasal mucosa was associated with the fungal spore and beta(1-->3)-glucan exposure (r(S) = 0.58 0.59, p < 0.05). CONCLUSION: These results are based on a relatively small population, and conclusions must be drawn with care. The results suggested that a moderate exposure to fungal spores, endotoxins, and beta(1-->3)-glucans during waste handling induced upper airway inflammation dominated by neutrophil infiltration and swelling of the nasal mucosa. PMID- 12771398 TI - Lung cancer mortality among chromate production workers. AB - AIMS: To assess mortality in 1997 among 493 former workers of a US chromate production plant employed for at least one year between 1940 and 1972. METHODS: Cohort members were followed for mortality to 31 December 1997. Standardised mortality ratios (SMRs) were calculated for selected cause specific categories of death including lung cancer. Lung cancer mortality was investigated further by calculation of SMRs stratified by year of hire, duration of employment, time since hire, and categories of cumulative exposure to Cr(VI). RESULTS: Including 51 deaths due to lung cancer, 303 deaths occurred. SMRs were significantly increased for all causes combined (SMR = 129), all cancers combined (SMR = 155), and lung cancer (SMR = 241). A trend test showed a strong relation between lung cancer mortality and cumulative hexavalent exposure. Lung cancer mortality was increased for the highest cumulative exposure categories (> or =1.05 to <2.70 mg/m(3)-years, SMR = 365; > or =2.70 to 23 mg/m(3)-years, SMR = 463), but not for the first three exposure groups. Significantly increased SMRs were also found for year of hire before 1960, 20 or more years of exposed employment, and latency of 20 or more years. CONCLUSIONS: The finding of an increased risk of lung cancer mortality associated with Cr(VI) exposure is consistent with previous reports. Stratified analysis of lung cancer mortality by cumulative exposure suggests a possible threshold effect, as risk is significantly increased only at exposure levels over 1.05 mg/m(3)-years. Though a threshold is consistent with published toxicological evidence, this finding must be interpreted cautiously because the data are also consistent with a linear dose response. PMID- 12771399 TI - Personal exposure assessment in the epidemiology of air pollutants. PMID- 12771400 TI - Predictions of mortality from mesothelioma. PMID- 12771401 TI - Mortality results for polyurethane manufacture understated. PMID- 12771402 TI - Triple gene block: modular design of a multifunctional machine for plant virus movement. AB - Many plant virus genera encode a 'triple gene block' (TGB), a specialized evolutionarily conserved gene module involved in the cell-to-cell and long distance movement of viruses. The TGB-based transport system exploits the co ordinated action of three polypeptides to deliver viral genomes to plasmodesmata and to accomplish virus entry into neighbouring cells. Although data obtained on both the TGB and well-studied single protein transport systems clearly demonstrate that plant viruses employ host cell pathways for intra- and intercellular trafficking of genomic nucleic acids and proteins, there is no integral picture of the details of molecular events during TGB-mediated virus movement. Undoubtedly, understanding the molecular basis of the concerted action of TGB-encoded proteins in transporting viral genomes from cell to cell should provide new insights into the general principles of movement protein function. This review describes the structure, phylogeny and expression of TGB proteins, their roles in virus cell-to-cell movement and potential influence on host antiviral defences. PMID- 12771403 TI - Use of Spring beauty latent virus to identify compatible interactions between bromovirus components required for virus infection. AB - Spring beauty latent virus (SBLV) is a member of the genus Bromovirus, and is closely related to Brome mosaic virus (BMV) and Cowpea chlorotic mottle virus (CCMV). Compatible interactions between viral components are required for successful infection of plants by BMV and CCMV. To further our understanding of interactions between bromovirus components, we used SBLV to produce reassortants among the three bromoviruses. We found that SBLV RNA 2 functioned with heterologous bromovirus RNA 1 in infections of whole plants and protoplasts of Nicotiana benthamiana, although SBLV RNA 1 did not function with heterologous bromovirus RNA 2. A DNA-based transient assay for 1a and 2a proteins, which are encoded by RNAs 1 and 2, respectively further suggested that SBLV 2a protein may function in combination with heterologous bromovirus 1a protein. Moreover, analysis of the ability of reassortants to spread locally revealed that an RNA 2 mediated interaction between viral components may be required for efficient cell to-cell movement of bromoviruses. PMID- 12771404 TI - Infectivity of recombinant strawberry vein banding virus DNA. AB - Infectivity of the cloned DNA genome of strawberry vein banding virus (SVBV) was demonstrated by particle bombardment of 4-week-old strawberry (Fragaria vesca L. var. UC-5) plants with gold particles coated with the putative full-length 7.9 kb viral DNA. Vein banding symptoms developed on 15 % of inoculated plants 6-7 weeks post-inoculation. An approximate 1.25-mer of the viral DNA was cloned into the binary vector pCGN1547. Particle bombardment of this construct into strawberry plants gave an infection rate of 75 %. The construct was used for transformation of Agrobacterium tumefaciens, and infiltration of these cells into healthy strawberry leaves resulted in development of vein banding symptoms in 100 % of inoculated plants. Gel electrophoresis, Southern blot hybridization with an SVBV probe and sequence analyses of PCR-amplified DNA fragments were used to confirm SVBV infection in symptomatic plants. PMID- 12771405 TI - Comparison of virus production in chicken embryo fibroblasts infected with the WR, IHD-J and MVA strains of vaccinia virus: IHD-J is most efficient in trans Golgi network wrapping and extracellular enveloped virus release. AB - Modified vaccinia virus Ankara (MVA) is an attenuated strain derived from vaccinia virus (VV) Ankara that grows efficiently in primary chicken embryo fibroblasts (CEFs) and baby hamster kidney cells only. MVA produces significantly more of the enveloped forms of VV in infected CEFs compared with VV strain Copenhagen. In the present study, production of the different infectious forms of VV was compared in CEFs infected with MVA or with two well-characterized replication-competent VV strains, WR and IHD-J. In a time-course experiment, the infectivity associated with the extracellular enveloped virus (EEV), the cell associated enveloped virus (CEV) and intracellular mature and enveloped viruses was determined. Further, the production of the different viral forms was quantified by electron microscopy (EM). The data collectively indicate that IHD-J is most efficient in producing all of the trans-Golgi network-wrapped forms and releases the highest titres of EEVs into the extracellular medium, with WR being least efficient. MVA initially replicated with faster kinetics, resulting in more intracellular virus and CEVs between 8 and 24 h post-infection (p.i.). As assessed by EM, the faster growth kinetics of MVA resulted in 3.5-fold more CEVs at the cell surface at 24 h p.i., compared with both WR and IHD-J. Accordingly, we found that despite the presence of two in-frame deletions in the A36R gene of MVA, this virus was able to make actin tails in CEFs. PMID- 12771406 TI - Phenotypic and genotypic characterization of clinical isolates of herpes simplex virus resistant to aciclovir. AB - A panel of 10 clinical isolates of herpes simplex virus (HSV) deficient in the expression of thymidine kinase (TK) and phenotypically resistant to aciclovir was characterized. Sequence analysis revealed a variety of mutations in TK (nucleotide substitutions, insertions and deletions), most of which resulted in truncated TK polypeptides. In line with previous reports, the most common mutation was a single G insertion in the 'G-string' motif. One HSV-1 isolate and two HSV-2 isolates appeared to encode full-length polypeptides and, in each case, an amino acid substitution likely to be responsible for the phenotype was identified. Pathogenicity was determined using a zosteriform model of HSV infection in BALB/c mice. The majority of isolates appeared to show impaired growth at the inoculation site compared with wild-type virus. They also showed poor replication in the peripheral nervous system and little evidence of zosteriform spread. One exception was isolate 4, which had a double G insertion in the G-string but, nevertheless, exhibited zosteriform spread. These studies confirmed that TK-deficient viruses display a range of neurovirulence with respect to latency and zosteriform spread. These results are discussed in the light of previous experience with TK-deficient viruses. PMID- 12771407 TI - Characterization of a neurovirulent aciclovir-resistant variant of herpes simplex virus. AB - A clinical isolate of herpes simplex virus type 1 that is aciclovir resistant but neurovirulent in mice was described previously. The mutation in this virus is a double G insertion in a run of seven G residues that has been shown previously to be a mutational hotspot. Using a sensitive assay, it has been demonstrated that preparations of this virus are able to induce low but consistent levels of thymidine kinase (TK) activity. However, this activity results from a high frequency mutational event that inserts a further G into the 'G-string' motif and thus restores the TK open reading frame. Passage of this virus through the nervous system of mice results in the rapid selection of the TK-positive variant. Thus, this variant is the major component in virus reactivated from latently infected ganglia. Mutation frequency appears to be influenced by the genetic background of the virus. PMID- 12771408 TI - The genome of herpesvirus papio 2 is closely related to the genomes of human herpes simplex viruses. AB - Infection of baboons (Papio species) with herpesvirus papio 2 (HVP-2) produces a disease that is clinically similar to herpes simplex virus (HSV-1 and HSV-2) infection of humans. The development of a primate model of simplexvirus infection based on HVP-2 would provide a powerful resource to study virus biology and test vaccine strategies. In order to characterize the molecular biology of HVP-2 and justify further development of this model system we have constructed a physical map of the HVP-2 genome. The results of these studies have identified the presence of 26 reading frames that closely resemble HSV homologues. Furthermore, the HVP-2 genome shares a collinear arrangement with the genome of HSV. These studies further validate the development of the HVP-2 model as a surrogate system to study the biology of HSV infections. PMID- 12771409 TI - Five unique open reading frames of infectious laryngotracheitis virus are expressed during infection but are dispensable for virus replication in cell culture. AB - The chicken alphaherpesvirus infectious laryngotracheitis virus (ILTV) exhibits several unique genetic features including an internal inversion of a conserved part of the unique long genome region. At one end, this inversion is preceded by a cluster of five open reading frames (ORFs) of 335-411 codons, designated ORF A to ORF E, that are not present in any other known herpesvirus genome. In this report we analysed expression of these genes and identified the corresponding viral RNA and protein products. Northern blot analyses showed 3'-coterminal transcripts of ORFs A and B, and monocistronic mRNAs of ORFs C and D. ORF E is part of a 3'-coterminal transcription unit that includes the conserved glycoprotein H and thymidine kinase genes. Monospecific antisera obtained after immunization of rabbits with bacterial fusion proteins allowed detection of the protein products of ORF A (40 kDa), ORF B (34 kDa), ORF C (38 and 30 kDa), ORF D (41 kDa) and ORF E (44 kDa) in ILTV-infected cells. For functional analyses, five virus recombinants possessing deletions within the individual ORFs and concomitant insertions of a reporter gene cassette encoding green fluorescent protein were generated. All virus mutants were replication competent in cell culture, but exhibited reduced virus titres or plaque sizes when compared to wild type ILTV. These findings indicate that the ILTV-specific ORF A to ORF E genes might be important for virus replication in the natural host organism. PMID- 12771410 TI - Genetic characterization of the unique short segment of phocid herpesvirus type 1 reveals close relationships among alphaherpesviruses of hosts of the order Carnivora. AB - To further characterize phocid herpesvirus type 1 (PhHV-1) at the molecular level, a cluster of genes comprising the complete unique short (Us) region of PhHV-1 has been cloned and sequenced. Within this region, ORFs were detected that code for the equivalent of the Us 2- protein of herpes simplex virus (HSV), a putative protein kinase, and for the glycoprotein equivalents gG, gD, gI and gE. In addition, two small ORFs downstream of gE, homologous to the Us 8.5 and Us 9 proteins of HSV were identified. Comparative analysis of the ORF encoding the gD equivalent of PhHV-1 identified the corresponding proteins of the alphaherpesviruses canine herpesvirus and, to lesser degree, feline herpesvirus as the closest relatives. PMID- 12771411 TI - Rescue of human cytomegalovirus strain AD169 tropism for both leukocytes and human endothelial cells. AB - Endothelial cell-tropism- and leukocyte- (polymorphonuclear- and monocyte-) tropism (leukotropism) are two important biological properties shared by all recent clinical isolates of human cytomegalovirus (HCMV). These properties are lost during extensive propagation of HCMV isolates in human fibroblasts, as shown by reference laboratory-adapted strains AD169 and Towne. Here we show that strain AD169 may reacquire both properties in vitro, endothelial (both venous and arterial) cell-tropism preceding leukotropism (predominantly involving monocytes). Restriction fragment length polymorphism analysis and sequencing performed on the original virus inoculum from human fibroblasts and serial passages on endothelial cells confirmed virus identity. Thus, fundamental biological properties may be lost and reacquired in vitro according to the cell culture system employed. The lack of a 15 kb DNA fragment in the strain AD169 genome does not prevent the rescue of these biological functions, thus indicating that they are likely to be encoded by viral genes located elsewhere. PMID- 12771412 TI - Human cytomegalovirus requires cellular deoxycytidylate deaminase for replication in quiescent cells. AB - We have previously observed that the expression of two thymidylate biosynthesis enzymes, dihydrofolate reductase and thymidylate synthase (TS), is upregulated in quiescent human fibroblasts infected with human cytomegalovirus (HCMV). Here, we have demonstrated that HCMV increases expression of the cellular deoxycytidylate deaminase (dCMP deaminase), which provides the substrate for TS by converting dCMP to dUMP. We observed an increase in dCMP deaminase protein levels, whereas deoxyuridine triphosphatase (dUTPase), another cellular enzyme that may provide dUMP by hydrolysing dUTP, was undetectable. The essential requirement of cellular dCMP deaminase for productive HCMV replication was further emphasized by showing that a precursor of a potent dCMP deaminase inhibitor, zebularine, suppressed virus replication and DNA synthesis. These results suggest that HCMV exploits the host's dCMP deaminase activity to replicate in quiescent cells. PMID- 12771413 TI - Updated Epstein-Barr virus (EBV) DNA sequence and analysis of a promoter for the BART (CST, BARF0) RNAs of EBV. AB - Two sequences required for activity of the Epstein-Barr virus BART RNA promoter in transfection assays have been identified by site-directed mutagenesis. One contains a consensus AP-1 site; the other has some similarity to Ets and Stat consensus binding sites. Candidate sequences were suggested by mapping a region of unmethylated DNA in EBV around the BART promoter followed by in vivo footprinting the promoter in the C666-1 nasopharyngeal carcinoma cell line, which expresses BART RNAs. The data are presented in the context of a revised EBV DNA sequence, known as EBV wt, that is proposed as a future standard sequence for EBV. PMID- 12771414 TI - The latency-associated nuclear antigen of Kaposi's sarcoma-associated herpesvirus interacts preferentially with the terminal repeats of the genome in vivo and this complex is sufficient for episomal DNA replication. AB - The genome of Kaposi's sarcoma-associated herpesvirus (KSHV) persists in latently infected cells as a circular episome. The latency-associated nuclear antigen (LANA) has been shown to tether viral DNA fragments to chromosomes and is proposed to maintain the KSHV genome. In order to identify the in vivo-binding sites for LANA on the whole KSHV genome and to analyse the function of this protein-DNA interaction, different in vivo systems have been developed. Chromatin immunoprecipitation experiments using three different cell lines latently infected with KSHV demonstrated that LANA binds preferentially and directly to the terminal repeats (TRs) but not to other regions of the viral chromosome in vivo. In contrast, in vitro LANA-DNA binding was much less specific. To identify autonomously replicating sequences within the KSHV genome, BCBL-1 cells were transfected with cosmids representing the entire genome. Cosmid Z2, consisting of the right end of the unique region and TRs, persisted as an episome in short-term assays. Long term, stable episome replication was observed with constructs derived from Z2 containing TRs only. LANA expression constructs containing a variable number of TRs replicated stably as episomes in uninfected cells. A 424 bp subfragment of the 801 bp TR could mediate episome replication. These studies show that LANA is a trans-acting protein that binds preferentially to TRs in vivo and these two elements are sufficient for episome replication. These results also suggest that the LANA expression plasmids reported here could be utilized as episomal vectors in a manner similar to Epstein-Barr virus-based vectors. PMID- 12771415 TI - The latency protein LANA2 from Kaposi's sarcoma-associated herpesvirus inhibits apoptosis induced by dsRNA-activated protein kinase but not RNase L activation. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) uses several strategies to counteract the interferon (IFN) system. In this study, the relationship of the protein LANA2 from KSHV to the IFN-activated protein kinase (PKR) and 2-5A system was analysed. It was found that LANA2 could not abrogate apoptosis or RNA degradation mediated by the 2-5A system. However, expression of LANA2 inhibited apoptosis triggered by PKR. LANA2 also counteracted the PKR-mediated inhibition of protein synthesis and partially blocked PKR-induced phosphorylation of eIF 2alpha. Analysis of PKR-induced activation of caspases 3 and 9 revealed that LANA2 abrogated activation of caspase 3 but not of caspase 9. These findings show that LANA2 is able to interfere with downstream events triggered by PKR. Hence, LANA2 should be considered as a KSHV defence protein against IFN. PMID- 12771416 TI - Transcription mapping of human herpesvirus 8 genes encoding viral interferon regulatory factors. AB - The human herpesvirus 8 (HHV-8) genome contains four tandemly arranged genes encoding viral interferon regulatory factors (vIRF-1 to 4) located between genes 57 and 58. Transcript mapping techniques were employed to determine the sizes, ends and splicing patterns of mRNAs specified by these genes in HHV-8-infected cell lines untreated or chemically induced into the lytic growth cycle. Depending on the cell line used, vIRF-3 transcription was minimally or not induced (i.e. expressed with latent kinetics), whereas the other vIRFs were inducible (i.e. expressed with lytic kinetics). Each gene possessed its own promoter (or promoters) and polyadenylation sites, and all but vIRF-1 were spliced from two exons. vIRF-1 was transcribed in uninduced and induced cells from a single initiation site preceded by a TATA box, with the possible use of an additional TATA box and initiation site in uninduced cells. In induced cells, vIRF-2 was transcribed from a single major initiation site preceded by a TATA box, and vIRF 4 was expressed from two sites each preceded by a TATA box. Transcripts for these genes were insufficiently abundant in uninduced cells to map the 5'-ends. vIRF-3 lacks an obvious TATA box and exhibited heterogeneous 5'-ends in uninduced and induced cells. These data clarify and extend our understanding of the structure and transcription of the HHV-8 vIRF genes. PMID- 12771418 TI - Human papillomavirus type 6 virus-like particles present overlapping yet distinct conformational epitopes. AB - The epitope for a human papillomavirus (HPV) type 6 conformation-dependent, neutralizing monoclonal antibody (mAb) was partially mapped using HPV L1 recombinant virus-like particles (VLPs). The mAb H6.J54 is cross-reactive with the closely related HPV types 6 and 11. By making HPV-6-like amino acid substitutions in the cottontail rabbit papillomavirus (CRPV) major capsid protein L1, we were able to transfer H6.J54 binding activity into a CRPV/HPV-6 hybrid L1 protein. Full binding activity was achieved with only nine amino acid changes and identified a region centred on the HPV-6 residues 49-54. This region has previously been shown to be a critical part of HPV-6 type-specific epitopes. Fine mapping of the region by scanning a series of alanine substitution mutations showed that in HPV-6 VLPs this type-common epitope overlaps HPV-6 type-specific epitopes. PMID- 12771417 TI - Glycoproteins M and N of human herpesvirus 8 form a complex and inhibit cell fusion. AB - Glycoproteins M (gM) and N (gN) are well conserved across the herpesvirus family and their involvement in virus penetration and egress is well described, especially for alphaherpesviruses. Because there was no previous study on the homologues of human herpesvirus 8 glycoproteins M (gM8) and N (gN8), we analysed their biochemical and functional characteristics. We found that: (i) gM8 aggregated following heat treatment; (ii) gM8 was a virion component; (iii) gM8 and gN8 were N-glycosylated; (iv) gM8 formed a specific complex with gN8; and (v) gN8 was required for functional processing of gM8. Co-expression of gM8 and gN8 inhibited cell fusion induced either by a combination of herpes simplex virus type 1 glycoproteins or by Molony murine leukaemia virus envelope protein. These results indicate that, in addition to the similar biochemical properties, the fusion inhibition reported previously only for alphaherpesviruses is a function conserved in the gammaherpesvirus subfamily. PMID- 12771419 TI - Seroepidemiology of the human polyomaviruses. AB - To assess the stability of polyomavirus antibodies in serial samples over time and the incidence and age-specific prevalence of polyomavirus infections, we established enzyme immunoassays (EIAs) using purified yeast-expressed virus-like particles (VLPs) containing the VP1 major capsid proteins of JC virus (JCV) and the AS and SB strains of BK virus (BKV). A random subsample of 150 Finnish women who had serum samples taken during the first trimester of pregnancy and had a second pregnancy during a 5 year follow-up period was selected, grouped by age of first pregnancy. The polyomavirus antibody levels were similar in samples taken during the first and second pregnancies (correlation coefficient 0.93 for BKV SB and 0.94 for JCV). Analysis of serum samples from 290 Swedish children aged 1-13 years, grouped by age in 2 year intervals, demonstrated that BKV seropositivity increased rapidly with increasing age of the children, reaching 98 % seroprevalence at 7-9 years of age, followed by a minor decrease. JCV seroprevalence increased only slowly with increasing age and reaching 72 % positivity among mothers >25 years of age. The age-specific seroprevalence of the human polyomaviruses measured using this VLP-based EIA was similar to previous serosurveys by other methods. The stability of the antibodies over time indicates that polyomavirus seropositivity is a valid marker of cumulative virus exposure, and polyoma VLP-based EIAs may therefore be useful for epidemiological studies of these viruses. PMID- 12771420 TI - Genetic variability of porcine parvovirus isolates revealed by analysis of partial sequences of the structural coding gene VP2. AB - The 3'-terminal 853 nt (and the putative 283 aa) sequence of the VP2-encoding gene from 29 field strains of porcine parvovirus (PPV) were determined and compared both to each other and with other published sequences. Sequences were examined using maximum-parsimony and statistical analyses for nucleotide diversity and sequence variability. Among the nucleotide sequences of the PPV field strains, 26 polymorphic sites were encountered; 22 polymorphic sites were detected in the putative amino acid sequence. Mapping polymorphic sites of protein data onto the three-dimensional (3D) structure of PPV VP2 revealed that almost all substitutions were located on the external surface of the viral capsid. Mapping amino acid substitutions to the alignment between PPV VP2 sequences and the 3D structure of canine parvovirus (CPV) capsid, many PPV substitutions were observed to map to regions of recognized antigenicity and/or to contain phenotypically important residues for CPV and other parvoviruses. In spite of the high sequence similarity, genetic analysis has shown the existence of at least two virus lineages among the samples. In conclusion, these results highlight the need for close surveillance on PPV genetic drift, with an assessment of its potential ability to modify the antigenic make-up of the virus. PMID- 12771421 TI - Transcriptional analysis of the white spot syndrome virus major virion protein genes. AB - White spot syndrome virus (WSSV) is a member of a new virus family (Nimaviridae) infecting crustaceans. The regulation of transcription of WSSV genes is largely unknown. Transcription of the major WSSV structural virion protein genes, vp28, vp26, vp24, vp19 and vp15, was studied to search for common promoter motifs for coordinate expression. The temporal expression of these genes and both 5' and 3' ends of the mRNA were determined, using infected crayfish gill tissue as a RNA source. RT-PCR showed that all five genes are expressed late in infection compared to the early ribonucleotide reductase large subunit gene. 5' RACE studies revealed a consensus late transcription initiation motif for only two of the five major virion protein genes. This motif was only found in one other upstream region of the putative translational start site of a gene with unknown function (ORF 158). No other conserved sequence motifs could be detected in the sequences surrounding the transcriptional start sites of the five major virion protein genes. All 5' ends were located about 25 nt downstream of an A/T rich sequence, including the consensus TATA-box sequence for vp15. The absence of a consensus motif is distinct from gene regulation of other large dsDNA viruses and suggests a unique regulation of WSSV transcription, in line with its unique taxonomic position. PMID- 12771422 TI - Amino acid mutations in the replicase protein nsP3 of Semliki Forest virus cumulatively affect neurovirulence. AB - It has been shown previously that an avirulent Semliki Forest virus (SFV) clone, rA774, engineered to carry the nsP3 gene of the virulent clone SFV4 becomes highly neurovirulent and is lethal for adult BALB/c mice. rA774, like several other alphaviruses, has an opal termination codon close to the 5' end of nsP3 (aa 469), while SFV4 has an arginine residue at this position. Mutation of the opal codon to an arginine residue increases the virulence of rA774 but does not reconstruct the severe neurovirulence of SFV4. Additionally, nsP3 amino acid sequences differ between these two strains by eight amino acids and by a deletion of seven amino acids in the C-terminal third of rA774 nsP3. This study shows that neurovirulence can be reconstituted gradually by exchanging individual amino acids and is fully retained when combinations of two nsP3 mutations, V(11)-->I and L(201)-->F, V(11)-->I and D(249)-->N, A(48)-->E and G(70)-->A or T(435)-->A and F(442)-->L, are introduced into an rA774 derivative carrying R(469). The critical role of the arginine codon for neurovirulence was confirmed further by the acquisition of a fully lethal phenotype following the introduction of R(469) into a moderately virulent rA774 recombinant carrying the SFV4 nsP1 and nsP2 genes. In conclusion, virulence determinants in SFV are distributed over a wide region of the nonstructural genes. PMID- 12771423 TI - The major envelope protein, GP5, of a European porcine reproductive and respiratory syndrome virus contains a neutralization epitope in its N-terminal ectodomain. AB - A set of neutralizing monoclonal antibodies (mAbs) directed against the GP(5) protein of European type porcine reproductive and respiratory syndrome virus (PRRSV) has been produced previously (Weiland et al., 1999). This set reacted with a plaque-purified virus (PPV) subpopulation of Dutch isolate Intervet-10 (I 10), but not with the European prototype PRRSV LV. In order to map the neutralization epitope in the GP(5) protein of the PPV strain, the ORF5 nucleotide sequence of PPV was determined. When the amino acid sequence derived from this nucleotide sequence was compared with that of PRRSV LV, four amino acid differences were found. Using site-directed mutagenesis, we showed that a proline residue at position 24 of the GP(5) sequence of the PPV strain enabled recognition by the neutralizing mAbs. Pepscan analysis demonstrated that the epitope recognized by the neutralizing mAbs stretched from residues 29 to 35. Surprisingly, the reactivity of the mAbs in the Pepscan system was independent of the presence of a proline in position 24. Moreover, residue 24 is located within the predicted signal peptide, implying that either the signal peptide is not cleaved or is cleaved due to the presence of Pro(24) such that the epitope remains intact. Our results demonstrate the presence of a neutralization epitope in the N-terminal ectodomain of the GP(5) protein of PRRSV and imply a role for the ectodomain of GP(5) in the infection of PRRSV. PMID- 12771424 TI - Characterization of the structure and variability of an internal region of hepatitis C virus RNA for M1 RNA guide sequence ribozyme targeting. AB - Accessibility to folded RNA and low potential of variation in the target RNA are crucial requirements for ribozyme therapy against virus infections. In hepatitis C virus (HCV), the sequence of the 5'UTR is conserved but the highly folded RNA structure severely limits the number of accessible sites. To expand investigation of targeting in the HCV genome, we have considered an internal genomic region whose sequence variation has been widely investigated and which has a particularly conserved RNA structure, which makes it accessible to the human RNase P in vitro. We have first mapped the accessibility of the genomic RNA to complementary DNAs within this internal genomic region. We performed a kinetic and thermodynamic study. Accordingly, we have designed and assayed four RNase P M1 RNA guide sequence ribozymes targeted to the selected sites. Considerations of RNA structural accessibility and sequence variation indicate that several target sites should be defined for simultaneous attack. PMID- 12771425 TI - Hepatitis C virus IRES efficiency is unaffected by the genomic RNA 3'NTR even in the presence of viral structural or non-structural proteins. AB - Hepatitis C virus (HCV) translation is mediated by an IRES structure. Instead of a poly(A) tail, the 3' end of the genome contains a tripartite 3'NTR composed of a non-conserved region, a polypyrimidine tract and a highly conserved stretch of 98 nt, termed the 3'X region. Using a set of bicistronic recombinant DNA constructs expressing two reporter genes separated by the HCV IRES, it was determined whether the HCV 3'NTR sequence, in the presence or absence of HCV proteins, played a role in the efficiency of HCV IRES-dependent translation ex vivo. Bicistronic expression cassettes were transfected into hepatic and non hepatic cell lines. These results show that neither the entire 3'NTR nor the 3'X sequence alters IRES-dependent translation efficiency, whatever the cell line tested. A potential effect of the 3'NTR on IRES-dependent translation in the presence of HCV proteins was investigated further. Neither non-structural nor structural HCV proteins had any effect on the efficiency of IRES in this system. In addition, in order to mimic HCV genome organization, monocistronic expression cassettes containing the IRES and a Core-DsRed fusion gene were constructed with or without the 3'NTR. In this context, no effect of the 3'NTR on IRES translation efficiency was observed, even in the presence of HCV proteins. These data demonstrate that HCV translation is not modulated by the viral genomic 3'NTR sequence, even in the presence of HCV structural or non-structural proteins. PMID- 12771426 TI - T helper responses to Japanese encephalitis virus infection are dependent on the route of inoculation and the strain of mouse used. AB - T helper cytokine and IgG subtype responses were studied in three strains of mice (C57BL/6J, Swiss albino, BALB/c; n=90 per strain) immunized with live Japanese encephalitis virus (JEV) by intraperitoneal (IP), subcutaneous (SC) and peroral (PO) routes. Lymphocytes obtained from the spleens of immunized and control mice were stimulated in vitro with JEV for 48 h and the supernatants were assayed for the presence of the cytokines IL-4 and IFN-gamma. JEV-specific IgG isotypes were also measured in the sera of immunized mice. T helper cytokine responses in mice immunized with JEV were found to be strain- and route-specific in the three species tested. Moreover, they were also dependent on the type of immunogen used (live vs killed virus), as well as the number of doses administered. C57BL/6J and BALB/c mice were more uniform in their T helper responses compared with the outbred Swiss albino mice and induced a good Th1 response (P<0.001). Among the three routes evaluated, the IP and SC routes consistently elicited a Th1 response compared with the PO route (P<0.001), where an initial Th2-type response reverted to a Th1 response after repeated immunization. Live JEV induced a Th1 response while the commercial killed vaccine induced a predominant Th2 profile. PMID- 12771427 TI - Molecular epidemiology of dengue virus type 3 in Venezuela. AB - During the past 40 years, dengue haemorrhagic fever and dengue shock syndrome (DHF/DSS) have emerged in humans, with approximately 3 million cases reported and over 58 000 deaths. Dengue virus serotypes 1, 2 and 4 (DENV-1, -2 and -4) have been co-circulating in Venezuela for at least the past 10 years, causing minor or major outbreaks of dengue fever (DF) and DHF/DSS. The first recorded outbreak due to DENV-3 in Venezuela dates to 1964 and the virus then seems to have disappeared. However, DENV-3 re-appeared recently (in July, 2000) in Venezuela after 32 years of absence and produced a prolonged major outbreak, which, by the end of 2001, involved 83 180 cases of dengue, mostly DF (92 %). Previous phylogenetic studies revealed that the DENV-3 circulating during the 1960s Latin American outbreak was a genotype V virus. To gain a better understanding of the nature of the current epidemic, the complete sequence was determined of the envelope (E) gene of 15 Venezuelan DENV-3 viruses isolated during 2000 and 2001 from patients presenting with different disease severity. Sequence data were used in phylogenetic comparisons with global samples of DENV-3. Analysis revealed that the strain circulating in Venezuela is closely related to isolates that were previously present in Panama and Nicaragua in 1994 and since then have spread through Central American countries and Mexico. This study also confirms previous reports showing that the DENV-3 strain currently circulating in the Americas is related to the strain that caused DHF epidemics in Sri Lanka and India in 1989 1991 (genotype III). Finally, no evidence of the re-emergence of the strain that circulated in Venezuela in the late 1960s and 1970s (genotype V) was found. PMID- 12771428 TI - Infection competition against grouper nervous necrosis virus by virus-like particles produced in Escherichia coli. AB - Dragon grouper (Epinephelus lanceolatus) nervous necrosis virus (DGNNV) comprises 180 copies of capsid protein that encapsulate a bipartite genome of single stranded (+)-RNAs. This study reports that virus-like particles (VLPs) are formed in Escherichia coli expressing the full-length ORF encoding the DGNNV capsid protein. Two sizes of VLPs are observed. The heavier particles resemble the native piscine nodavirus in size and stain permeability, while the lighter ones are approximately two-thirds of the full size. The recombinant VLPs block attachment of native virus to the surface of cultured fish nerve cells, blocking infection by the native virus. PMID- 12771429 TI - Comparisons of the complete genomes of Asian, African and European isolates of a recent foot-and-mouth disease virus type O pandemic strain (PanAsia). AB - During the last 12 years, a strain of foot-and-mouth disease (FMD) virus serotype O, named PanAsia, has spread from India throughout Southern Asia and the Middle East. During 2000, this strain caused outbreaks in the Republic of Korea, Japan, Russia (Primorsky Territory), Mongolia and South Africa (KwaZulu-Natal Province), areas which last experienced FMD outbreaks in 1934, 1908, 1964, 1974 and 1957, respectively. In February 2001, the PanAsia strain spread to the United Kingdom where, in just over 7 months, it caused outbreaks on 2030 farms. From the UK, it quickly spread to the Republic of Ireland, France and the Netherlands. Previous studies that utilized RT-PCR to sequence the VP1-coding region of the RNA genomes of approximately 30 PanAsia isolates demonstrated that the UK virus was most closely related to the virus from South Africa (99.7 % nucleotide identity). To determine if there was an obvious genetic reason for the apparently high level of fitness of this new strain, and to further analyse the relationships between the PanAsia viruses and other FMDVs, complete genomes were amplified using long-range PCR techniques and the PCR products were sequenced, revealing the sequences for the entire genomes of five PanAsia isolates as well as an animal-passaged derivative of one of them. These genomes were compared to two other PanAsia genomes. These analyses revealed that all portions of the genomes of these isolates are highly conserved and provided confirmation of the close relationship between the viruses responsible for the South Africa and UK outbreaks, but failed to identify any genetic characteristic that could account for the unprecedented spread of this strain. PMID- 12771430 TI - The implications of virus diversity within the SAT 2 serotype for control of foot and-mouth disease in sub-Saharan Africa. AB - SAT 2 is the serotype most often associated with outbreaks of foot-and-mouth disease (FMD) in livestock in southern and western Africa and is the only SAT type to have been recorded outside the African continent in the last decade. Its epidemiology is complicated by the presence of African buffalo (Syncerus caffer), which play an important role in virus maintenance and transmission. To assess the level of genetic complexity of this serotype among viruses associated with both domestic livestock and wildlife, complete VP1 gene sequences of 53 viruses from 17 countries and three different host species were analysed. Phylogenetic analysis revealed eleven virus lineages, differing from each other by at least 20 % in pairwise nucleotide comparisons, four of which fall within the southern African region, two in West Africa and the remaining five in central and East Africa. No evidence of recombination between these lineages was detected, and thus we conclude that these are independently evolving virus lineages which occur primarily in discrete geographical localities in accordance with the FMD virus topotype concept. Applied to the whole phylogeny, rates of nucleotide substitution are significantly different between topotypes, but most individual topotypes evolve in accordance with a molecular clock at an average rate of approximately 0.002 substitutions per site per year. This study provides an indication of the intratypic complexity of the SAT 2 serotype at the continental level and emphasizes the value of molecular characterization of diverse FMD field strains for tracing the origin of outbreaks. PMID- 12771431 TI - Mapping epitopes in equine rhinitis A virus VP1 recognized by antibodies elicited in response to infection of the natural host. AB - Equine rhinitis A virus (ERAV) is an important respiratory pathogen of horses and is of additional interest because of its close relationship and common classification with foot-and-mouth disease virus (FMDV). As is the case with FMDV, the VP1 capsid protein of ERAV has been shown to be a target of neutralizing antibodies. In FMDV VP1, such antibodies commonly recognize linear epitopes present in the betaG-betaH loop region. To map linear B cell epitopes in ERAV VP1, overlapping fragments spanning its length were expressed in Escherichia coli as glutathione S-transferase (GST) fusion proteins. These fusion proteins were tested for reactivity with sera from ERAV-infected horses and with polyclonal sera from ERAV-immunized rabbits and mice. Regions at the N- and C termini as well as the betaE-betaF and the betaG-betaH loop regions contained B cell epitopes that elicited antibodies in the natural host. GST fusion proteins of these regions also elicited antibodies following immunization of rabbits and mice, which, in general, strongly recognized native ERAV VP1 but which were non neutralizing. It is concluded that the N-terminal region of ERAV VP1, in particular, contains strong B cell epitopes. PMID- 12771432 TI - Rabies virus matrix protein regulates the balance of virus transcription and replication. AB - RNA synthesis by negative-strand RNA viruses (NSVs) involves transcription of subgenomic mRNAs and replication of ribonucleoprotein complexes. In this study, the envelope matrix (M) protein of rabies virus (RV) was identified as a factor which inhibits transcription and stimulates replication. Transcription, but not replication, of RV minigenomes or of full-length RV was decreased by expression of heterologous M. Since RV assembly involving M and the glycoprotein G renders virus synthetically quiescent, an RV was generated with the M and G genes substituted by placeholders. Surprisingly, RNA synthesis by this recombinant was characterized not only by an increased transcription rate but also by a diminished accumulation of replication products. This phenotype was reversed in a dose-dependent manner by providing M in trans, showing that M is a replication stimulatory factor. The role of M in determining the balance of replication and transcription was further exploited by generation of a recombinant RV with attenuated M expression, which is highly active in transcription. Regulation of RNA synthesis by matrix proteins may represent a general mechanism of nonsegmented NSVs, which is probably obscured by the silencing activity of M during virus assembly. PMID- 12771433 TI - An immunostimulatory oligodeoxynucleotide containing a cytidine-guanosine motif protects senescence-accelerated mice from lethal influenza virus by augmenting the T helper type 1 response. AB - The SAM-P1 strain of senescence-accelerated model mice shows an impaired T helper type 1 (Th1) immune response upon infection with influenza virus, which results in high susceptibility to the virus. Treatment of spleen cells from SAM-P1 mice with an immunostimulatory oligodeoxynucleotide containing a cytidine-guanosine motif (CpG ODN) in vitro increased the ratio of the titre of IFN-gamma to that of IL-4. Administration of CpG ODN to SAM-P1 mice generated satisfactory virus specific cytotoxic T-lymphocyte responses and natural killer cell activation and the virus-specific immunoglobulin (Ig) isotype switched from IgG1 to IgG2a. Virus growth in the lungs of CpG ODN-treated SAM-P1 mice was cleared quickly and mice survived the lethal influenza virus infection. It could be inferred that a possible mechanism of CpG ODN for normalization of senescence-associated dysregulation of the Th1/Th2 balance involves the upregulated expression of CD154 and CD40 molecules on immune-competent cells. These results suggest that CpG ODN could contribute to the development of a protective strategy against infectious diseases, especially among immunocompromised elderly persons, by stimulating Th1 immune responses. PMID- 12771434 TI - Evidence that avian reovirus sigmaA protein is an inhibitor of the double stranded RNA-dependent protein kinase. AB - The results of a previous study demonstrated that avian reovirus is highly resistant to the antiviral effects of interferon and suggested that the double stranded RNA (dsRNA)-binding sigmaA protein might play an important role in that resistance. To gather more evidence on the interferon-inhibitory activity of sigmaA protein, its gene was cloned into the prokaryotic maltose-binding protein (MBP) gene fusion vector pMalC and into the recombinant vaccinia virus WRS2. The two recombinant sigmaA proteins displayed a dsRNA-binding affinity similar to that of sigmaA protein synthesized in avian reovirus-infected cells. Interestingly, MBP-sigmaA, but not MBP, was able to relieve the translation inhibitory activity of dsRNA in reticulocyte lysates by blocking the activation of endogenous dsRNA-dependent enzymes. In addition, transient expression of sigmaA protein in HeLa cells rescued gene expression of a vaccinia virus mutant lacking the E3L gene, and insertion of the sigmaA-encoding gene into vaccinia virus conferred protection for the virus against interferon in chicken cells. Further studies demonstrated that expression of recombinant sigmaA in mammalian cells interfered with dsRNA-dependent protein kinase (PKR) function. From these results we conclude that sigmaA is capable of reversing the interferon-induced antiviral state by down-regulating PKR activity in a manner similar to other virus-encoded dsRNA-binding proteins. PMID- 12771435 TI - Mutational analysis of two zinc-finger motifs in the nucleocapsid protein of simian immunodeficiency virus mac239. AB - To clarify the physiological function of two zinc-finger (ZF) motifs in the nucleocapsid (NC) protein of simian immunodeficiency virus (SIV), we constructed three mutant viruses with alterations in either or both motifs using a molecular clone of SIVmac (SIVmac239). An immunoblot analysis of the cell lysates transfected with DNA mutated in either the first (ZF1) or second (ZF2) motif showed that the amount of partially processed Gag products (Pr46) was greater than that produced by the wild-type (WT). The genomic RNA contents in the viral particles released from the transfected cells were measured by quantitative RT PCR. Values for the ZF1 and ZF2 mutants and the double mutant were 26, 20 and 7 % that of the WT, respectively, indicating that the two ZF motifs of SIVmac239 NC protein function almost equivalently with respect to RNA encapsidation and processing of Gag precursors. Despite the presence of some genomic RNA in the mutant viruses, they lost all viral infectivity. To determine the reason for this, we examined (using PCR) to which step viral DNA synthesis proceeded in the mutant viruses. We did not see any block up to the step of minus-strand DNA synthesis. However, plus-strand DNA synthesis after plus-strand transfer did not occur in any of the mutant viruses. These findings indicated that the mutations in the ZF motifs of SIVmac led to a loss of infectivity due partly to impairment of DNA synthesis, in addition to inefficient encapsidation of genomic RNA. PMID- 12771436 TI - Isotype control of B cell signaling. AB - The B cell receptor (BCR) consists of an antigen-binding membrane immunoglobulin (mIg) associated with the CD79alpha and CD79beta heterodimer. Naive B cells express the IgM and IgD isotypes, which have very short cytoplasmic tails and therefore depend on CD79alpha and CD79beta for signal transduction. After antigenic stimulation, B cells undergo isotype switching to yield IgG, IgE, or IgA. Recent research suggests that the ability of the B cell coreceptor CD22 to regulate BCR signaling depends on the isotype of the mIg cytoplasmic tail. Cell lines that express a BCR with the cytoplasmic tail from IgG, the isotype found in memory B cells, are not subject to CD22 regulation, whereas cell lines that express BCRs with IgM cytoplasmic tails are subject to CD22 regulation. Moreover, stimulation through BCRs containing an IgG cytoplasmic tail causes increased numbers of antigen-specific clones to accumulate. These observations are a valuable step toward understanding the difference in B cell signaling between na ve and memory cells. Here, we discuss the implications of these findings for CD22 regulation and signaling through the mIgG-containing BCR. PMID- 12771437 TI - Cryofibrinogenaemia complicated by amyloidosis: a new association. PMID- 12771438 TI - Triple primary sarcoma in Werner syndrome with a novel mutation. PMID- 12771439 TI - Haemophagocytic syndrome in a rheumatoid arthritis patient treated with infliximab. PMID- 12771440 TI - Chronic pulmonary toxicity of methotrexate and rheumatoid arthritis. PMID- 12771442 TI - Development of systemic lupus erythematosus in a young child affected with chronic granulomatous disease following withdrawal of treatment with interferon gamma. PMID- 12771443 TI - Antioxidant status and lipid peroxidation in seminal plasma and spermatozoa of patients with ankylosing spondylitis. PMID- 12771444 TI - The high prevalence of low bone density in men aged 55 yr and over presenting with low trauma fractures to an accident and emergency department. PMID- 12771445 TI - Pseudotumour cerebri presentation of systemic lupus erythematosus: more than an association. PMID- 12771446 TI - Arthritis as a cause of prolonged in-patient admission in acute medical beds. PMID- 12771447 TI - Screening for extensor tendon rupture in rheumatoid arthritis. PMID- 12771448 TI - Frequency and causes of osteoporosis in men. PMID- 12771450 TI - Invisible microscopic endometriosis: how wrong is the sampson hypothesis of retrograde menstruation to explain the pathogenesis of endometriosis? PMID- 12771451 TI - 'Invisible' microscopic endometriosis: a review. AB - Endometriosis can have very subtle visual appearances. All theories of origin imply some early and presumably tiny form of the disease which potentially cannot be detected by the unaided human eye. A review of the literature indicates that with increasing magnification at surgery, virtually all endometriosis can be visualized. Invisible microscopic endometriosis is an unproven entity which retards intellectual progress in the study of this disease. PMID- 12771453 TI - On the use of classical naturopathy and complementary medicine procedures in hospitals and clinics practicing gynecology and obstetrics in Germany. Results of a questionnaire survey. AB - BACKGROUND: Classical naturopathy and complementary medicine procedures are used in all industrial countries, the only difference being the extent to which they are used. However, information pertaining to effects, safety and costs of such therapies remains incomplete. An important aim of this study was to establish to what extent women's hospitals and clinics in Germany adopt complementary medicine and naturopathy as a means of treatment and how often they are used and by whom. MATERIALS AND METHODS: In a questionnaire regarding the use of complementary medicine and naturopathy, we contacted 1054 gynecology/obstetrics institutions. Of the 1054 contacted, 481 (45.6%) of the returned questionnaires could be analyzed and evaluated using descriptive statistical methods. RESULTS: Altogether per clinic an average of four different complementary medicine therapies was named which were used in the field of gynecology and obstetrics. Acupuncture (used in 94.1% of responding clinics) and homeopathy (used in 83.0% of responding clinics) were without doubt the two procedures that were used most often. Clear differences regarding both choice of therapy and frequency of use of a given therapy could be seen when comparing areas of the former German Democratic Republic (GDR) with those of the former Federal Republic of Germany (FRG). No differences could be found pertaining to the comparison of university and non university hospitals. CONCLUSION: Interesting tendencies could be seen regarding the willingness of hospital departments to widen their therapy spectrum by adopting complementary medicine procedures. These in turn led us to valuable conclusions as to the various strategies of patient care. Given the frequency of use of complementary medicine therapies in the field of gynecology and obstetrics, it must surely be concluded that research into complementary medicine approaches in such areas must be intensified. PMID- 12771452 TI - 'In vitro' cytokine secretion by peripheral blood and decidual lymphocytes during the third trimester of normal pregnancy. AB - If the 'Th2 phenomenon' is dependent on trophoblastic antigens and cytokines, the profile of cytokines secreted by decidual lymphocytes (DL) should indicate stronger Th2 shift than that of peripheral blood lymphocytes (PBL). We studied spontaneous and mitogen-stimulated 'in vitro' cytokine secretion of cultured lymphocytes isolated from peripheral blood (n = 21) and decidua (n = 11) of third trimester healthy pregnant women not being in labor. The ELISA method was used for estimation of IL-2, IL-4, IL-6, IL-10, IL-12, IFN-gamma and TGF-beta. The results (given in pg/ml) were found to be statistically significant in the Mann Whitney U-test (p < 0.05). Compared to PBL, DL produced spontaneously decreased concentrations of IL-12, IL-6, IFN-gamma and TGF-beta and upon mitogen stimulation increased quantities of IL-2, IL-4 and IL-10. It demonstrates that DL play an active role in local Th2-like immunoregulation and that their cytokine secretion pattern estimated in 'in vitro' conditions is biased towards Th2 activity more than that of PBL. PMID- 12771454 TI - The ALUI calcitonin receptor gene polymorphism (TT) is associated with low bone mineral density and susceptibility to osteoporosis in postmenopausal women. AB - Osteoporosis is a common disorder with a strong genetic component. Our aim was to evaluate the correlation of the ALUI calcitonin receptor gene polymorphism to bone mineral density and their relationship to osteoporosis. We determined the ALUI calcitonin receptor gene polymorphism using polymerase chain reaction-based restriction analysis in 167 postmenopausal women in Taiwan. The polymorphism was detected by the restriction enzyme ALUI, where the C allele indicated the absence of the cuttable site and the T allele indicated its presence. Bone mineral density of the lumbar spine and proximal femur were measured using dual-energy X ray absorptiometry. The allelic frequencies for the 167 postmenopausal women in Taiwan were 86.5% for C and 13.5% for T in ALUI restriction fragment length polymorphisms. The prevalence of each genotype in the study population was 2.4% TT, 22.2% CT, and 75.4% CC. The three genotypic groups differed significantly in unadjusted and adjusted bone mineral density at the lumbar spine and the femoral neck. Unadjusted and adjusted bone mineral density values were lowest in women with the TT genotype. The ALUI calcitonin receptor genotype showed a positive association with prevalence of osteoporosis in the subjects. That is, women with genotype TT had a greater risk for developing osteoporosis at the lumbar spine and at the femoral neck. The ALUI calcitonin receptor gene polymorphism is associated with reduced bone mineral density and predisposes women to osteoporosis, but should be interpreted with caution because of the small number of subjects in the unfavorable TT genotype. PMID- 12771455 TI - Effect of two modes of antiandrogen treatment on insulin sensitivity and serum leptin in women with PCOS. AB - Androgens are suggested to interact with leptin production and with insulin sensitivity in both polycystic ovary syndrome (PCOS) and obesity. The aim of the study was to follow these interactions along with two forms of antiandrogen treatment. Twenty women with PCOS were treated with ethinylestradiol and high dose of cyproteroneacetate (EE-CA) and 8 with the gonadotrophin-releasing hormone (GnRH) analogue goserelin for 6 months. The patients were divided into a low and a high body weight group and compared with a group of overweight women without PCOS. Both treatments resulted in a significant reduction of free testosterone but the concentration of leptin remained unchanged. EECA treatment resulted in deterioration and GnRH in improvement of insulin sensitivity. Serum leptin correlated only with body weight and body fat. It is concluded that leptin levels do not adequately reflect changes in insulin sensitivity or androgen levels after short-term antiandrogen or antigonadotropin treatment. PMID- 12771456 TI - GnRH analogues, transvaginal ultrasound-guided drainage and intracystic injection of recombinant interleukin-2 in the treatment of endometriosis. AB - We performed a double-blind, randomised controlled trial to evaluate the results of ultrasound-guided aspiration of endometriomas under the effect of GnRH analogues and a possible additional beneficial effect by leaving 600000 IU of recombinant interleukin-2 (rIL-2) in the cysts. Twenty-four women with endometriosis-related symptoms, increased values of CA-125 and transvaginal ultrasonography showing endometriomas >3 cm who were initially sent to us for laparotomy and conservative surgery for endometriosis were included. Main outcome measures were severity of symptoms, size and percentage of echographical reduction of endometriomas and CA-125 levels after 2 menses post-GnRH analogues. Secondary outcome measures were the time until recurrence of abnormal parameters and the need for surgery after treatment. We found moderate clinical results after treatment with drainage plus GnRH analogues and significantly improved results in women having received rIL-2 intracystically. There were no side effects. Two out of 3 previously infertile patients became pregnant after therapy. Though the rates of recurrence of endometriomas >or=3 cm were similar in both groups, the period until recurrence was significantly greater when rIL-2 was used, and the rates of recurrence of symptoms and increased CA-125 values were also significantly lower in patients who received rIL-2. Surgery was finally performed on 10 patients (4 with and 6 without previous rIL-2 treatment) during follow-up (30 +/- 12.7 months). These findings led to the conclusion that transvaginal ultrasound-guided puncture and aspiration of endometriomas under endometrial suppressive therapy with GnRH analogues have some value for endometriosis treatment, improving the clinical manifestations and avoiding some surgical therapies, and that rIL-2 left in the cyst increases these beneficial effects significantly. PMID- 12771457 TI - Laparoscopically assisted vaginal hysterectomy versus total abdominal hysterectomy: a study of 100 cases on light-endorsed transvaginal section. AB - The objective of this study was to compare the results of a modified laparoscopically assisted vaginal hysterectomy (LAVH) procedure, using light endorsed transvaginal section by two puncture trocars, with those of total abdominal hysterectomy (TAH) in a prospective, randomized, short-term study. A new, modified LAVH technique using Endo GIA stapler and two puncture trocars was established. For the laparoscopic phase, each adnexum was dissected, and the vesicouterine junction was identified clearly with the laparoscopic light from the vaginal side. Vaginal-phase surgery was performed as usual. Two hundred patients scheduled for abdominal hysterectomy were randomized to either LAVH (n = 100) or TAH (n = 100). Duration of hospitalization, time of surgery, dose of analgesics, and rates of complications were significantly lower in the LAVH group (p < 0.001). The average operating time was 77 +/- 30 min for LAVH and 102 +/- 18 min for TAH. The duration of hospitalization was 3.2 +/- 0.7 days for LAVH and 5.5 +/- 1.3 days for TAH. There were three complications in the LAVH group and 15 in the TAH group. Postoperative meperidine requirements (1.2 vs. 3.7 ampoules, 1 ampoule = 50 mg) were significantly fewer in the LAVH group. Regarding the training time, the mean operating time in the first 20 cases was 98 min, and in the last 20 cases it was 70.9 min. As compared with TAH and other modified LAVH procedures reported previously, the present technique is easy to learn and timesaving with fewer complications. PMID- 12771458 TI - Surface area of the human vagina as measured from vinyl polysiloxane casts. AB - Sixty-two vinyl polysiloxane casts from subjects in the lying, supine position were measured for surface area with five shapes: conical, parallel sides, heart, slug, and pumpkin seed being included. The surface areas ranged from 65.73 to 107.07 cm(2) with a mean of 87.46 cm(2) and a standard deviation of 7.80 cm(2). There were no significant differences in surface area among the shape groups. And no correlations with surface area were found by parity or by rod length, introital diameter, posterior length of casts as measured with string, or greatest width and greatest depth of casts. 70% of the casts had a surface area of 84.50 cm(2) or above, and 90% had a surface area of 77.72 cm(2) or above. Variation within the interquartile range (75th to 25th percentile) was +/-5.2% of the median. PMID- 12771459 TI - Two episodes of hemoperitoneum from luteal cysts rupture in a patient with congenital factor X deficiency. AB - The clinical manifestation of two episodes of hemoperitoneum from ruptured corpus luteum cysts, during the luteal phase of the cycle in a young patient with the rare congenital factor X deficiency, is reported for the first time in literature. The correct diagnosis of the underlying disorder, the gynecological management and the regular follow-up can minimize the risks of this potentially life-threatening hematological disorder. PMID- 12771460 TI - Primary omental pregnancy presenting with hemorrhagic shock. A case report. AB - Abdominal pregnancy is a very rare and serious type of extrauterine gestation that accounts for approximately 0.003% of all ectopic pregnancies. Omental pregnancy, an extremely rare form of abdominal pregnancy, can be primary or secondary to a tubal pregnancy that aborts out of the fimbria and reimplants in the peritoneal cavity. We present an additional case of primary omental pregnancy at 12 gestational weeks presenting with symptoms of hemoperitoneum and acute abdomen. PMID- 12771461 TI - Regulation of vascular endothelial growth factor by tamoxifen in vitro and in vivo. AB - OBJECTIVE: To evaluate the effects of tamoxifen on human endometrial vascular endothelial growth factor (VEGF) gene expression in vitro and in vivo. STUDY DESIGN: Ishikawa cells were transiently co-transfected with plasmids expressing human estrogen receptor (ER)-alpha or -beta and a 2.3-kb human VEGF promoter luciferase construct. Transfected cells were treated with tamoxifen and the increase in luciferase activity was compared to controls. In addition, immunohistochemical analysis of VEGF expression in endometrial biopsies from 11 postmenopausal women receiving tamoxifen was compared to 24 postmenopausal women on no hormone replacement. RESULTS: In transfected cells expressing ERalpha, tamoxifen induced a significant VEGF promoter activation (3.2-fold, p < 0.01), whereas cells expressing ERbeta showed a 1.6-fold stimulation (p = NS). No significant differences in VEGF immunostaining were observed in tamoxifen-exposed women. CONCLUSIONS: Although tamoxifen exposure increases VEGF expression in vitro, a significant increase in VEGF expression is not apparent in endometrial biopsies of postmenopausal women treated with this drug. PMID- 12771462 TI - An unusual case of bilateral ureteric obstruction after anterior colporrhaphy and vaginal hysterectomy for pelvic organ prolapse. AB - We report a case of bilateral ureteric obstruction after anterior colporrhaphy. The excessive folding of the bladder trigone after anterior colporrhaphy led to occlusion of both ureteric orifices. PMID- 12771463 TI - [Oral and written language production in prematures children: results in 7 1/2 year-old]. AB - Prematurity entails risk factors of cognitive delay, particularly in language production and reading. These risk factors may be related to physiological and/or environmental variables. This study has two main objectives: (1) to track down which preterm children are most at risk of developing cognitive delays, particularly language production and reading; (2) to study whether screening of oral language in 3 1/2-year-olds predicts their future reading ability. A cohort of 50 pre-term children (23 boys and 27 girls) was followed from the ages of 2 to 7 1/2 years. The results show that: (a) 28% of the premature children present delayed language production at 3 1/2 years of age; (b) 34% of these children present delayed reading at 7 1/2 years of age; (c) language production at 3 1/2 years predicts the reading ability at 7 1/2 years, and (d) premature children who are most at risk are those who belong to low sociocultural groups. PMID- 12771464 TI - An automatic method to quantify the vibration properties of human vocal folds via videokymography. AB - The study offers an automatical quantitative method to obtain vibration properties of human vocal folds via videokymography. The presented method is based on image processing, which combines an active contour model with a genetic algorithm to improve detecting precision and processing speed, can accurately extract the vibration wave in videokymograms and quantify the vibration properties in terms of eight typical parameters automatically. To verify the precision of the proposed algorithm, an indirect simulation setup of vocal folds has been performed. The verification result shows that the relative error of the entire simulation system is less than 5%. Applying the method to analyzing hundreds of videokymograms from 12 subjects, the result indicates that the vibration characteristics of vocal folds can be recognized more exactly, and diseases of the vocal folds can be diagnosed quantitatively. PMID- 12771465 TI - [Intrinsic and co-intrinsic vowel characteristics in cerebellar and parkinsonian dysarthrias]. AB - Few studies were made on the intrinsic and co-intrinsic vowel characteristics in neurologic dysarthrias. This work evaluates these parameters in a group of 4 subjects with cerebellar dysarthria suffering from a cerebellar degenerative syndrome and a group of 4 subjects with Parkinson disease suffering from parkinsonian dysarthria. These subjects were compared with 10 control subjects. An intensive speech treatment inspired from the Lee Silverman voice treatment (LSVT) was applied to one of the parkinsonian subjects of the study, with assessment before and after rehabilitation. The acoustic measures assessed the duration and the frequency of the vowels [a, i, ul indifferent contexts as well as their formant frequency and sentence duration. The results show different impairments according to the type of the dysarthria, and some forms of contrast transposition. Speech therapy restores subnormal contrasts. PMID- 12771466 TI - A comparison of voice quality ratings made by Japanese and American listeners using the GRBAS scale. AB - The GRBAS scale is a widely used method for perceptual evaluation of voice quality. Two linguistically diverse groups of listeners (Japanese and American) rated 35 voice samples using the GRBAS scale. The ratings obtained from the two groups were compared to determine if the different linguistic background affected the use of the GRBAS scale. Results show that there are no significant differences between the Japanese and American listeners in the use of the Grade, Roughness and Breathiness scales. Ratings on the Asthenia and Strain scales, however, were different between the two groups of listeners. Despite these discrepancies, the GRBAS scale may be an excellent tool for perceptual evaluation of voice quality by linguistically diverse groups. PMID- 12771467 TI - Enhancement of the depigmenting effect of hydroquinone and 4-hydroxyanisole by all-trans-retinoic acid (tretinoin): the impairment of glutathione-dependent cytoprotection? AB - Many of the well-known depigmenting agents such as hydroquinone and 4 hydroxyanisole are, in fact, melanocytotoxic chemicals which are oxidized in melanocytes to produce highly toxic compounds such as quinones. These cytotoxic compounds are responsible for the destruction of pigment cells, which results in skin depigmentation. However, cells are capable of protecting themselves against cytotoxic agents by intracellular glutathione (GSH). This protection takes place under the enzymatic action of the detoxification enzyme glutathione S-transferase (GST), which is responsible for the conjugation of toxic species to GSH. The depigmenting effect of hydroquinone is shown to be potentiated by buthionine sulfoximine (BSO) and cystamine as the result of the reduction of intracellular levels of GSH by these two agents. Additionally, BSO and cystamine are shown to inhibit the activity of GST. The combination of all-trans-retinoic acid (tretinoin, TRA) with hydroquinone or 4-hydroxyanisole is also known to produce synergetic skin depigmentation. TRA serves as a potent inhibitor of mammalian GSTs and is known to make cells more susceptible to the cytotoxic effect of chemicals by inhibiting the activity of this enzyme. This agent is also shown to reduce the level of intracellular GSH in certain cells. We have proposed that the mechanism of action of TRA to synergistically enhance the melanocytotoxic effect of chemicals involves the inhibition of GST and the impairment of glutathione dependent cytoprotection against melanocytotoxic agents. PMID- 12771468 TI - Effect of lesion size on the diagnostic performance of dermoscopy in melanoma detection. AB - BACKGROUND: Dermoscopy is able to correctly classify a higher number of melanomas than naked-eye examination. Little is known however about factors which may influence the diagnostic performance during practice. The aim of the study was to analyze the effect of size of the lesion on diagnostic performance of dermoscopy in melanoma detection. METHODS: Eight dermatologists examined clinical and, separately, clinical and dermoscopic (combined examination) images of 200 melanocytic lesions previously excised [64 melanomas, 24 in situ and 40 invasive (median thickness 0.30 mm) and 136 melanocytic nevi]. After examination, diagnostic performance was analyzed in accordance with the major diameter of the lesions divided into 3 groups, i.e. small (less than 6 mm), intermediate (between 6 and 9 mm) and large (10 mm or more) lesions. These groups were shown to be highly comparable concerning the microstaging of melanomas (median thickness value 0.30, 0.22 and 0.32 mm, respectively). RESULTS: Dermoscopy increased the diagnostic performance of naked-eye examination of both intermediate and large lesions [sensitivity value: +19.3 (p = 0.002) and +10.3 (p = 0.007); diagnostic accuracy value: +7.4 (p = 0.004) and +6.1 (p = 0.07)]. On the contrary, no statistically significant increase was found dealing with small lesions (sensitivity +3.7, p = 0.66; diagnostic accuracy -1.7, p = 0.55). CONCLUSIONS: The diagnostic improvement associated with the addition of dermoscopy to naked eye examination is influenced by the size of the lesion, i.e. it is lacking with lesions up to 6 mm in diameter. The optimized use of dermoscopy in melanoma detection is obtained dealing with melanocytic lesions 6 mm in diameter or larger. PMID- 12771469 TI - Association of urticaria pigmentosa with cafe-au-lait spots, neurofibromas and neurofibroma-like neoplasms: a mere coincidence? AB - BACKGROUND: Urticaria pigmentosa (UP) is characterized by dense aggregates of mast cells in the dermis. There is consistent evidence from the literature that mast cells may play a pathogenetic role in the development of neurofibromas and other tumors. OBJECTIVE: To study the concomitant appearance of UP with neurofibromas and neurofibroma-like neoplasms. METHODS: We analyzed 31,752 records of patients examined at the Department of Dermatology in the year 2000, looking for UP and associated neurofibromas and neurofibroma-like neoplasms in persons younger than 18 years. RESULTS: We identified a total of 27 patients suffering from UP, with 16 persons younger than 18 years. One 12-year-old Caucasian boy demonstrated multiple cutaneous mastocytomas consistent with the diagnosis of UP. On his trunk, four cafe-au-lait spots were found. A cutaneous neurofibroma was confirmed by skin biopsy. Magnetic resonance imaging detected multiple neoplasms located at the nerve roots of the spine, resembling plexiform neurofibromas. CONCLUSIONS: There may be a concomitant appearance of UP and neoplasms, with mast cells possibly playing a causative role. The existence of neoplasms, including neurofibromas and neurofibroma-like lesions should be considered when examining UP cases. PMID- 12771470 TI - Lupus-erythematosus-like eruption induced by Trichophyton mentagrophytes infection. AB - BACKGROUND: Several factors are known to trigger acute manifestations of lupus erythematosus (drugs, ultraviolet radiation, bacterial or viral infections, pregnancy), but fungal infections have never been reported to induce lupus-like lesions. We describe 2 children with tinea capitis caused by Trichophyton mentagrophytes(TM), who developed transient skin and serological manifestations of systemic lupus erythematosus. PATIENTS: Case 1, a 3-year-old girl, had a kerion due to TM transmitted by an octodon, and a facial skin eruption suggestive of systemic lupus erythematosus. Antinuclear antibodies (ANA) were positive at 1:250. After griseofulvin treatment, the lupus-like rash completely regressed, and the ANA titre decreased. Case 2, a 4-year-old girl, had occipital kerion and an annular scaly erythema on the shoulder caused by TM. She also had a non-scaly rash on the face and a recent history of photosensitivity. The ANA titre was positive at 1:8,000. Cutaneous lesions disappeared after an 8-week griseofulvin therapy, and ANA titres decreased progressively. CONCLUSIONS: 'New pets' are usually the vectors of TM, especially familiar rodents like the degu of Chile (also named octodon). In our 2 cases, lupus-like rashes began before the onset of griseofulvin treatment, suggesting a pathogenic role of the dermatophyte. A wide variety of lesions named 'mycides' was described several decades ago associated with TM infections. We hypothesize that these mycides and the TM-induced lupus like lesions reported in our 2 cases could represent the same entity. PMID- 12771471 TI - Intramuscular alefacept improves health-related quality of life in patients with chronic plaque psoriasis. AB - BACKGROUND: Psoriasis profoundly affects patient quality of life (QOL). Amevive (alefacept), a novel and selective biologic agent, was clinically effective and significantly improved QOL in a phase 2 study. METHODS: The present placebo controlled, randomized phase 3 study examined the effects of a 12-week course of intramuscular alefacept on QOL in 507 patients with chronic plaque psoriasis using both dermatology-specific questionnaires [Dermatology Life Quality Index (DLQI); Dermatology Quality of Life Scales (DQOLS)] and the Short Form-36 Health Survey (SF-36), a generic, general health questionnaire. RESULTS: All 3 QOL measures (DLQI, DQOLS, SF-36) demonstrated that alefacept 15 mg was significantly more effective than placebo in improving QOL in patients with chronic plaque psoriasis. In addition, the improvements in QOL for patients in the alefacept 15 mg group were of similar magnitude for all 3 QOL measures. CONCLUSION: The findings of this study are an important addition to the QOL literature for psoriasis treatments. PMID- 12771472 TI - A randomized, double-blind, placebo-controlled trial of vitamin C iontophoresis in melasma. AB - BACKGROUND: Vitamin C is known to both inhibit melanin formation and reduce oxidized melanin. However, vitamin C does not easily penetrate the skin. In this study, vitamin C iontophoresis was employed in order to enhance vitamin C penetration. OBJECTIVE: The purpose of this study was to evaluate the efficacy of vitamin C iontophoresis for melasma patients. METHODS: Twenty-nine females with melasma were enrolled. For iontophoresis, a vitamin C solution was applied to one side of the face, while distilled water was applied to the other side as a control. The L (luminance) value was measured by a colorimeter to obtain an objective pigmentation parameter. RESULTS: Twelve weeks after iontophoresis, the colorimeter of the treated site showed a significant decrease in the L value (from 4.60 to 2.78, p = 0.002), compared to that of the control site (from 4.45 to 3.87, p = 0.142). CONCLUSION: Vitamin C iontophoresis may be an effective treatment modality for melasma. PMID- 12771473 TI - Type 2 segmental manifestation of congenital multiple glomangiomas. AB - Congenital multiple glomus tumors are extremely rare, and less than 20 cases have been well documented. We report an uncommon case of congenital multiple glomangiomas with a segmental manifestation in a 9-year-old girl. Since birth, the child had presented asymptomatic angiomatous macules arranged in a segmental pattern on the neck and trunk. During a follow-up period of 9 years, disseminated smaller papulonodular lesions developed on both arms and the left leg with a segmental distribution. Histopathology of congenital and acquired lesions confirmed the diagnosis of glomangiomas. The family history was negative for glomus tumors. This type of presentation supports the recently described type 2 segmental manifestation of multiple glomus tumors, with a segmental involvement of congenital early developing lesions. PMID- 12771474 TI - Papulonodular mucinosis associated with subacute cutaneous lupus erythematosus. AB - Mucin deposition is a common histopathologic finding in lupus erythematosus (LE) but is rarely present in sufficient quantity to produce clinically apparent skin lesions. Until now, fewer than 40 cases of papulonodular mucinosis associated with LE have been reported in the literature, and it was associated with either systemic LE or discoid LE. For the first time, to our knowledge, we describe 2 patients with subacute cutaneous LE who presented with papulonodular mucinosis as a major clinical manifestation of their disease. PMID- 12771476 TI - Transformation between Kaposiform hemangioendothelioma and tufted angioma. AB - Kaposiform hemangioendothelioma and tufted angioma are viewed as two separate disease entities in the current classification of vascular tumors. However, the concurrence of features of both vascular tumors in the same specimen of some patients raises the question whether these tumors exist on a continuum. Herein, we report a dynamic transformation between both tumors within a single patient, which suggests they are two variants of the same vascular tumor. PMID- 12771475 TI - Primary cutaneous CD30+ large T-cell lymphoma in a patient with psoriasis treated with cyclosporine. AB - We report the case of a 61-year-old woman who developed an anaplastic CD30+ cutaneous T-cell lymphoma during oral cyclosporine (CsA) therapy for recalcitrant psoriasis. Two months after CsA discontinuation, clinical and histological resolution of the lymphoma was observed. However, 3 years later extracutaneous involvement of the lymphoma could be detected. The association between CsA administration and the occurrence of the lymphoma may be casual, but a relationship with immunosuppression may also be hypothesized. We have reviewed all relevant data in the literature. To our knowledge, this is the first case of primary cutaneous CD30+ anaplastic large T-cell lymphoma in a patient treated with CsA for psoriasis. PMID- 12771477 TI - Recurrent E413K mutation of hHb6 in a Japanese family with monilethrix. AB - Monilethrix is an autosomal dominant hair disorder characterized by a beaded appearance of the hair due to periodic thinning of the shaft. This disorder has been reported to be caused by mutations in the helix termination motif of two type II cortex keratins, hHb1 and hHb6. Here we describe a Japanese monilethrix family that has the most frequent mutation, the E413K mutation in hHb6, so far found in 26 families. Genotype/phenotype correlation was not obvious in our case or in the previously reported cases. PMID- 12771478 TI - Allergic contact dermatitis from etofenamate without cross-sensitization to other anthranilic acid derivatives. PMID- 12771479 TI - Acute idiopathic scrotal edema in children. PMID- 12771480 TI - Malignant melanoma in a tattoo. PMID- 12771481 TI - Subacute lichenoid eruption due to L-thyroxine overdosage. PMID- 12771482 TI - Short-contact therapy with tazarotene in psoriasis vulgaris. PMID- 12771483 TI - Epper syndrome: an additional case. PMID- 12771484 TI - Nail destruction in pemphigus vulgaris. AB - Involvement of the nails in pemphigus vulgaris (PV) is rare and is usually seen when the disease is severe. The most common clinical manifestations are chronic paronychia and onychomadesis. Finger nails are more frequently involved than toe nails. We report a case of severe and persistent PV in which an exacerbation was preceded by an erosive and destructive lesion of the right hallux nail unit. A nail bed biopsy was performed to rule out a subungual tumour. The image of suprabasal acantholysis yielded the diagnosis of PV. This localisation and clinical manifestation of PV have only been reported once. PMID- 12771485 TI - Management of drug rash with eosinophilia and systemic symptoms (DRESS syndrome): an update. AB - Drug rash with eosinophilia and systemic symptoms (DRESS) syndrome is a rather distinct severe adverse drug reaction characterised by skin rash, fever, lymph node enlargement and internal organ involvement. Our aim was to review the available data regarding the management of this probably underrecognised subset of drug reaction. So far, the only undisputed way to treat severe hypersensitivity reactions is prompt withdrawal of the offending drug. The use of systemic corticosteroids remains controversial. The benefit of therapies aimed at accelerating the elimination of the causative drug deserves further studies. In the absence of a well-established therapy, primary and secondary prevention have a key role in the management of DRESS syndrome. PMID- 12771486 TI - Nudging epidermal field cancerogenesis by imiquimod. AB - BACKGROUND: The coexistence of malignancies close to each other on the skin has been occasionally reported. The concept of field cancerogenesis applies to such cases. Given the purported mechanism of action of imiquimod, it should not be surprising that this treatment could inhibit epidermal field cancerogenesis. AIM: To assess the effect of imiquimod applied twice weekly on incipient bowenoid changes disclosed in the vicinity of basal cell carcinomas. MATERIALS AND METHOD: Biopsies were taken before treatment and after 4-6 weeks and 12 weeks of imiquimod treatment. RESULTS: Large atypical bowenoid keratinocytes and dyskeratotic cells were cleared in time while factor-XIIIa-positive dermal dendrocytes appeared boosted and admixed with a brisk lymphocytic infiltration. CONCLUSION: Epidermal field cancerogenesis appears to be controlled by imiquimod. Dermal dendrocytes might play a pivotal role in this regression phenomenon. PMID- 12771487 TI - Prolonged imiquimod treatment and graft-versus-host reaction: histological mimicry in the skin infiltration pattern of the monocyte-macrophage-dendrocyte lineage. AB - Factor-XIIIa-positive dendrocytes belong to the monocyte-macrophage-dendrocyte (MMD) lineage which is considered to play a pivotal role in the skin response to the immune response modifier imiquimod. The same cells are also boosted in low grade graft-versus-host reaction (GVHR) with skin manifestations. Both conditions are characterized by specific epidermal damage. The aim of the present study was to compare them using immunohistochemistry to identify MMD subsets and other inflammatory cells in the dermis. We compared 3 cases of long-term (4, 9 and 11 months) imiquimod topical applications on normal skin, 25 low-grade GVHR controlled by immunosuppressive therapy and 25 control cases with normal skin. Compared to the normal dermis, cells of the MMD lineage were considerably boosted in the dermis of GVHR and at the imiquimod-treated sites. By contrast, only minimal accumulations of lymphocytes and Langerhans cells were disclosed in the dermis. The pattern of dermal infiltration by MMD cells was similar in GVHR and after imiquimod treatment. However, intraindividual differences in densities were obvious irrespective of the skin condition. In conclusion, there is great mimicry in the MMD involvement in the dermis during low-grade GVHR and after chronic applications of imiquimod. PMID- 12771488 TI - Histometric assessment of psoriatic plaques treated by vitamin D3 derivatives. AB - BACKGROUND: Psoriasis is an immunogenetic disorder. Factor XIIIa+ dermal dendrocytes (DD) are part of the pathobiological changes in the plaque type of the disease. OBJECTIVE: The present study aimed at comparing the effect of 3 vitamin D(3) derivatives on the epidermis, microvasculature and DD in psoriasis. METHOD: Twenty men suffering from chronic plaques of psoriasis on the trunk were enrolled in this study. They applied twice a day for 3 weeks calcipotriol, tacalcitol and calcitriol, each to one plaque. Another similar lesion received petrolatum as a placebo treatment. Skin biopsies were taken at entry and at completion of the 3-week treatment phase. Immunohistochemistry was performed using the lectin of Ulex europaeus and an antibody to factor XIIIa. Computerized image analysis served to measure the stratum Malpighii area, the microvasculature area and the DD numerical density in the papillary dermis. RESULTS: At entry in the study, the 4 test sites were indistinguishable with regard to the stratum Malpighii area, the papillary microvasculature area and the papillary DD density. The 3 histometric parameters appeared correlated with each other. At completion of the 3-week treatment phase, the 3 vitamin D derivatives had decreased the size of the stratum Malpighii. In addition, calcitriol had also reduced the DD density in the papillary dermis. No other significant changes were yielded. CONCLUSION: As assessed by histometry, the psoriatic epidermis responded to a short treatment using the 3 vitamin D derivatives. The better result compared to the control site was achieved by calcitriol. DD appeared to be most controlled by the same drug. The microvasculature did not appear to be decreased at the 3-week time point in treatment. PMID- 12771489 TI - Early treatment of hemangiomas with lasers. A review. AB - Hemangiomas can present a difficult problem in management. Much controversy exists over whether it is better to watch and wait for natural involution or to be more aggressive and attempt to prevent some of the potential negative sequelae. Different modalities have been employed in the treatment of hemangiomas, including systemic therapy - antiangiogenic drugs, i.e. systemic corticosteroids and alpha-interferon - and local therapy - surgical procedures, arterial embolization and cryotherapy. Overall, a very conservative approach to therapy has been recommended because of treatment risks, treatment inadequacy and lack of evidence showing superiority over natural involution. Recently laser treatment has been used in the therapeutic approach of hemangiomas. This review discusses the pros and contras of early laser treatment of hemangiomas. For superficial hemangiomas, the flashlamp pumped pulsed dye laser in particular has proven itself in numerous studies. In the treatment of hemangiomas with subcutaneous components, the Nd:YAG laser is the treatment of choice. PMID- 12771490 TI - Factor-XIIIa-positive dendrocytes in drug-induced toxic epidermal necrolysis (Lyell's syndrome): paradoxical activation in skin and rarefaction in lymph nodes. AB - BACKGROUND: Drug-induced toxic epidermal necrolysis (TEN) is a rare life threatening disease characterized by the extensive destruction of the epidermis contrasting with the discreteness of lymphoid cell infiltration. The precise pathomechanism of the disease remains unclear. METHODS: Skin specimens and peripheral and thoracic lymph nodes (LNs) were collected from 2 TEN patients. They were examined by conventional histology and immunohistochemistry using antibodies directed to factor XIIIa (dermal dendrocytes), CD1a (Langerhans cells), CD15 (granulocytes), CD20 (B lymphocytes), CD45RO (activated T lymphocytes), CD68 (macrophages) and the proliferation marker Ki-67. LNs from patients with mycosis fungoides, from subjects dead from acute cardiac failure or traumatic injuries, as well as metastasis-free sentinel LNs of cutaneous melanoma served as positive and negative controls. RESULTS: TEN LNs showed absence of germinal centers but a moderate hyperplasia of the paracortical T cell zone. Immunohistochemistry did not reveal any distinctive aspect in LN cellular densities between TEN and other control conditions except for the factor-XIIIa+ dendritic cells which were dramatically reduced in numbers in TEN LNs. This rarefaction in LNs contrasted with the great number of these cells in the skin of the same patients. CONCLUSION: The structure of TEN LNs rules out the involvement of an antibody-mediated response. As dendritic factor-XIIIa+ cells are involved in most cutaneous T-cell-mediated responses, their depletion in TEN LNs could explain the sparse lymphoid cell infiltrate in lesional TEN skin. PMID- 12771491 TI - Two cases of cross-sensitivity in subjects allergic to paraphenylenediamine following ingestion of Polaronil. AB - We report the cases of 2 women presenting allergy to paraphenylenediamine (PPD). Both patients had a history of eczema that worsened following the ingestion of the antihistamine Polaronil (dexamethasone/dexchlorpheniramine). This clinical presentation could be explained by cross-sensitivity to sulfanilic acid (4 aminobenzene sulfonic acid), a metabolite of sunset yellow (FD&C No. 6). Sunset yellow is an azo dye present in this tablet. Indeed, PPD-allergic subjects may suffer from cross-sensitivity to related compounds, especially to those that can be ingested such as azo dyes. Such compounds are used in some instances by the food and pharmaceutical industries, but their presence is often undisclosed. PMID- 12771492 TI - Thalidomide in refractory vulvar ulcerations associated with Crohn's disease. AB - We report a case of recurrent vulvar ulcerations developed on vegetations associated with Crohn's disease. Lesions responded to low-dose thalidomide treatment. A biopsy revealed a neutrophilic infiltrate with rare giant cells. Vulvar manifestations were consistent with metastatic Crohn's disease or with a reactive neutrophilic dermatitis. Thalidomide reduces the activity of tumor necrosis factor (TNF) alpha by accelerating the degradation of its mRNA. Nowadays, it has become a promising drug in a broad variety of disorders in which TNF-alpha seems to play a pivotal role, including inflammatory bowel diseases. This is the first case report presenting the efficacy of thalidomide in the treatment of vulvar involvement in Crohn's disease. PMID- 12771493 TI - Tinea capitis in Brussels: epidemiology and new management strategy. AB - BACKGROUND: We found pre-established directives inadequate to cope with the current increase in anthropophilic tinea capitis in Brussels. OBJECTIVES: To study new epidemiological profiles and to define new strategies for management and prevention. PATIENTS AND METHODS: A total of 122 children affected by tinea capitis were followed in our department from October 1, 2001, until September 30, 2002. The results were assessed retrospectively. RESULTS: Anthropophilic tinea capitis represented 89.34% of the cases. The implicated anthropophilic dermatophytes were by decreasing frequency: Microsporum langeronii (39.34%), Trichophyton soudanense (28.69%), Trichophyton violaceum (18.03%) and Trichophyton tonsurans (3.28%). CONCLUSION: The responsible pathogens reflect immigration flows originating mostly from Black and North Africa. Precise recommendations for each visit are detailed. PMID- 12771494 TI - Adverse skin reactions to anti-TNF-alpha monoclonal antibody therapy. AB - Various adverse cutaneous reactions to anti-TNF-alpha monoclonal antibody have been reported. In clinical studies with infliximab (Remicade) adverse drug reactions were most frequently reported in the respiratory system and in the skin and appendages. We describe here 6 patients receiving anti- TNF-alpha therapy (infliximab) for Crohn's disease or rheumatoid arthritis who consulted our out patient department for adverse cutaneous reactions between November 1999 and February 2002. The following diagnoses were made: leukocytoclastic vasculitis, lichenoid drug reaction, perniosis-like eruption (2 patients), superficial granuloma annulare and acute folliculitis. PMID- 12771495 TI - Lassueur-Graham Little-Piccardi syndrome: a 20-year follow-up. AB - Lassueur-Graham Little-Piccardi (LGLP) syndrome is a distinctive entity associating a scarring patchy alopecia of the scalp, a non-cicatricial axillary and pubic hair loss and a lichenoid follicular eruption. We present a 20-year follow-up of a demonstrative case, which had begun as typical lichen planus. In the literature, the relationship with lichen planus remains unclear. The diagnosis of LGLP syndrome should be evoked for all scarring alopecias. PMID- 12771496 TI - Granulomatous slack skin: treatment with extensive surgery and review of the literature. AB - We report an 11-year follow-up of a case of granulomatous slack skin. The patient was first treated surgically followed by a rapid relapse. Then he was treated by alpha-interferon during 15 months. When this treatment was stopped, the disease relapsed again. Extensive surgery was undertaken. Recently, a new relapse has occurred which was again treated by extensive surgery. No other manifestation of a lymphoproliferative disorder appeared. The clinical, histological, immunohistochemical, ultrastructural, molecular biological and genetic studies with a review of 39 other cases described in the literature are presented. PMID- 12771497 TI - Framing the future of antifungals in atopic dermatitis. AB - Atopic dermatitis (AD) is a frequent chronic inflammatory skin disease. Some fungal colonization or infection of the skin may exacerbate AD severity, particularly the so-called head and neck variant. In addition, excessive intestinal colonization by Candida albicans may represent an additional triggering factor. Hence, there is a rationale to use antifungals in selected AD patients. Early trials with topical ketoconazole in head and neck AD showed a decrease in Malassezia colonization, but no significant improvement was observed in the clinical severity. In contrast, clinical improvement and decreased serum IgE were obtained in patients with positive Malassezia radioallergosorbent tests (RASTs) who were treated by oral ketoconazole. Some preliminary data suggested that oral itraconazole treatment in AD patients reduced the need for topical corticosteroids, provided clinical improvement particularly in head and neck AD, reduced the cutaneous and intestinal fungal colonization that may trigger AD, reduced the percentage of positive Malassezia cultures and demonstrated a decrease in C. albicans and Malassezia RAST values. Furthermore, beside its antifungal action, itraconazole in part relieves pruritus and inflammation. In conclusion, oral itraconazole treatment can alleviate AD severity in selected patients. Fluconazole is also effective. Further research is warranted to identify whether the load in skin surface fungal agents, the fungal RAST values and specific prick testing should be assessed in order to optimize the antifungal management in AD patients. PMID- 12771498 TI - Longitudinal study of the three-dimensional development of subsurface enamel lesions during in vitro demineralisation. AB - A longitudinal study was made of the 3D development of subsurface enamel lesions in whole human molars. X-ray microtomography (XMT) was used to measure the 3D distribution of linear attenuation coefficients in the tissue at 8-15 stages during cumulative times of 36-107 days demineralisation through approximately 1 mm-wide windows. Although lesion morphology was consistent with preferential anisotropic dissolution parallel to enamel prisms at the advancing front, detail (at a scale of approximately 100 microm) varied in relation to exposed surface sites separated by <1 mm. The distribution of mineral in the most superficial region varied across the exposed face of each lesion. Within lesions, localised foci of low mineral concentration (at a scale of approximately 200 microm) retained their general form through successive stages of demineralisation before coalescing. The most advanced regions within a lesion seemed to correspond with surface regions with lowest mineral concentration. These findings indicate that local variations in fractional pore volume of partially demineralised enamel influence the subsequent spatial development of lesions. PMID- 12771499 TI - Six-year success rates of occlusal amalgam and glass-ionomer restorations placed using three minimal intervention approaches. AB - The present randomised clinical trial was aimed at comparing three minimally invasive restorative treatment approaches for managing dental caries in occlusal surfaces using a non-gamma-2 amalgam and a low-viscosity glass-ionomer as the restorative material. The treatment approaches tested in parallel groups were: conventional in a university setting, modified-conventional and ultraconservative (Atraumatic Restorative Treatment, ART) approaches in a field setting. A split mouth design was used in which the two restorative materials were randomly placed in 430 matched contralateral pairs of permanent molar teeth. A total of 152 children from five primary schools were recruited and treated by a dental therapist. The restorations were evaluated after 6 years by 2 calibrated independent examiners. The 6-year successes for all occlusal amalgam and glass ionomer restorations were 72.6 and 72.3%, respectively. There were no statistically significant differences observed between the successes for both amalgam and glass-ionomer restorations placed either by the ART (68.6%, with 95% CI = 61-76%) approach or by the conventional (74.5%, with 95% CI = 65-82%) and the modified-conventional (75.8%, with 95% CI = 67-83%) approaches after 6 years. There was also no statistically significant difference observed between the successes of occlusal ART restorations with glass-ionomer (67.1%, with 95% CI = 56-77%) and occlusal conventional restorations with amalgam (74%, with 95% CI = 61-85%) after 6 years. 'Restoration fracture/marginal defects' and 'loss of material' were the most common causes for failure. The former was more often recorded in amalgam restorations and the latter in glass-ionomer restorations. Secondary caries was observed for 2% of glass-ionomer and for 10% of amalgam restorations. This difference was statistically significant (p = 0.001). The ART approach using glass-ionomer performed equally well as conventional restorative approaches using electrically driven equipment and amalgam for treating dentinal lesions in occlusal surfaces after 6 years. PMID- 12771500 TI - In vivo cariostatic effect of black copper cement on carious dentine. AB - This study compared the effect of a copper phosphate cement (BCC) and a conventional glass ionomer cement (GIC) on carious dentine that remains under restorations in vivo. Using a split-mouth design, 45 primary molar pairs with dentine caries were sampled microbiologically. Without further removal of carious dentine, the molar pairs were randomly allocated to three restorative groups: (1) one cavity was lined with BCC and restored with GIC and the other was kept under review as an untreated control; (2) one cavity was restored with GIC, whilst the other was kept under review; (3) one cavity was lined with BCC and restored with GIC, whilst the other was filled with GIC. The dentine was re-sampled microbiologically at 1 month (30 pairs) and 6 months (15 pairs). BCC demonstrated a significant effect on the total anaerobic count over 1 month, when paired with both the control and GIC, whereas the antibacterial effects of GIC compared with no treatment were not statistically significant. In addition, BCC performed significantly better than no treatment in reducing mutans streptococci and lactobacilli over 1 month. Over 6 months, BCC caused a significantly greater reduction in mutans streptococci than GIC. In conclusion, BCC demonstrated a significant antibacterial effect on carious dentine in vivo. PMID- 12771501 TI - Non-mutans streptococci in patients receiving radiotherapy in the head and neck area. AB - OBJECTIVE: To study mutans and non-mutans streptococci in patients after radiotherapy of the head and neck. METHODS: Oral rinse samples collected from nasopharyngeal carcinoma patients before and after radiotherapy were diluted and cultured on nonselective and selective media for enumeration of total cultivable plaque flora, mutans and non-mutans streptococci and lactobacilli. Non-mutans streptococci were identified biochemically and by 16S rDNA sequence homology analysis. RESULTS: After irradiation, mutans streptococci were not isolated; the levels of Streptococcus mitis and lactobacilli increased significantly. The level of Streptococcus salivarius increased, but the significance was the borderline. The level of Streptococcus sanguis decreased significantly after irradiation. The abundance of other oral streptococci species showed no significant changes. CONCLUSIONS: S. mitis and S. salivarius are the predominant non-mutans streptococci in the high-caries-risk oral flora following radiotherapy. PMID- 12771502 TI - Routine replacement of tunneled, cuffed, hemodialysis catheters eliminates paraspinal/vertebral infections in patients with catheter-associated bacteremia. AB - BACKGROUND: Management of tunneled, cuffed, central venous catheters in hemodialysis (HD) patients with bacteremia remains a challenge. Attempts to salvage the catheter with systemic antibiotics alone have been associated with increased risk of metastatic infectious complications. METHODS: Retrospective case series of patients with infectious complications in a chronic dialysis unit, affiliated with a tertiary care university hospital. RESULTS: Between 1996 and 1999, when we treated HD catheter-associated bacteremia with systemic antibiotics alone, we encountered a clustering of 8 cases of paraspinal/vertebral infections in a population of 162 patients. After changing our protocol, i.e., routine catheter exchange over a guide wire at approximately 48 h, while on systemic antibiotic therapy, we did not encounter any new cases of paraspinal/vertebral infections over a 15-month period. CONCLUSION: Our experience suggests that routine exchange of tunneled, cuffed catheters over a guide wire in HD patients presenting with bacteremia may significantly reduce serious infectious complications, e.g., epidural abscess/vertebral osteomyelitis. PMID- 12771503 TI - Direct contact between human peripheral blood mononuclear cells and renal fibroblasts facilitates the expression of monocyte chemoattractant protein-1. AB - BACKGROUND: Cell-to-cell interaction is thought to be an important feature of a variety of biological processes. As far as the proinflammatory process is concerned, the interaction between mesangial cells and monocytes/macrophages induces the expression of monocyte chemoattractant protein-1 (MCP-1), and this may play a role in glomerulonephritis. In this study, we investigated whether the cell-to-cell interaction between immune cells and renal fibroblasts induces MCP-1 gene expression, which may be involved in interstitial inflammation in the kidney. METHODS: Human renal fibroblast cell lines, tNKF (from a normal kidney) and tFKIF (from a kidney with fibrosis), and peripheral blood mononuclear cells (PBMC) were used to assess the effect of cell-to-cell contact on the expression of MCP-1 mRNA in the fibroblasts. The expression of the MCP-1 gene in the fibroblasts was also examined after stimulation with tumor necrosis factor-alpha (TNF-alpha) and the culture supernatant from PBMC. RT-PCR was used to detect MCP 1 mRNA expression. Neutralizing antibodies to intercellular adhesion molecule-1 (ICAM-1) and vascular endothelial adhesion molecule-1 (VCAM-1) were used to block the cell-to-cell contact between the fibroblasts and PBMC. RESULTS: TNF-alpha and the culture supernatant from PBMC increased MCP-1 gene expression in tNKF cells. Contact culture with PBMC also significantly increased MCP-1 gene expression in tNKF cells. Although the basal level of MCP-1 mRNA was higher in tFKIF than tNKF cells, tFKIF cells did not respond significantly to any stimulation in this study. Following pretreatment with anti-ICAM-1 antibody, MCP-1 gene expression in tNKF cells was significantly suppressed in contact culture with PBMC. Anti-VCAM-1 antibody treatment had no effects. CONCLUSION: It is suggested that the interaction between renal fibroblasts and PBMC was mediated through direct contact and by secreted humoral factors. ICAM-1 on renal fibroblasts may be involved in the direct cell-to-cell interaction inducing MCP-1 gene expression, which seems to be involved in renal interstitial inflammation. PMID- 12771504 TI - Animal models for asthma: controversial aspects and unsolved problems. AB - The incidence of asthma in children and adults is continuously increasing in the developed countries. Its pathophysiology is still not yet clarified in sufficient detail. Despite a large number of animal models for asthma (mainly rodents), these only partly mimic the situation in humans. Some examples of limitations of using the mouse as an animal model for asthma will be discussed and directions of future research outlined. PMID- 12771505 TI - Endotoxins and allergy: lessons from the murine model. AB - Exposure early in life to organic dusts containing immunomodulatory components such as endotoxins and immunizing components such as aeroallergens may greatly influence whether subsequent encounters with allergens lead rather to sensitization or unresponsiveness. We investigated the effects of endotoxin in the context of allergen-mediated immune responses in a murine model of allergen sensitization. Systemic sensitization with ovalbumin induced high serum levels of allergen-specific IgE, predominant Th2-type cytokine production, eosinophilic airway inflammation and in vivo airway hyperreactivity. Endotoxins were either applied systemically prior to sensitization, or via the airways prior to airway challenges, or by repeated inhalation during the first weeks of life prior to subsequent sensitization. Different effects of endotoxins on allergen-induced immune responses may be attributed to differences in dosing, route of application, time relationship with allergen sensitization and the concurrent exposure to endotoxin and allergen. The results of these studies may help to define the effects of endotoxin on allergen-mediated immune reactions and to further delineate the important interrelationships between environment and disease development. Finally, this may lead to new strategies in the prevention and treatment of allergic diseases. PMID- 12771507 TI - Stimulation of bronchus-associated lymphoid tissue in rats by repeated inhalation of aerosolized lipopeptide MALP-2. AB - OBJECTIVE: Bronchus-associated lymphoid tissue (BALT) is a part of the integrated mucosal immune system. It may play an important functional role for antigen uptake and induction of specific immune reactions. The aim of this study was to investigate whether it is possible to induce or modulate BALT by the repetitive inhalation of the synthetic lipopeptide MALP-2. METHODS: Female Lewis rats (245 +/- 19 g) inhaled 25 microg of MALP-2 six times at intervals of 1 week. One week after the last inhalation, they were sacrificed. Cells of the bronchoalveolar lavage and the left lung were investigated by flow cytometry. The middle lobe of the right lung was embedded in paraffin. BALT was semiquantitatively measured in 15 serial cross sections per animal. RESULTS: After repetitive inhalation of the diluent as well as MALP-2, BALT was found. The total area was increased after repetitive treatment with MALP-2. In addition, the preferential incidence of BALT was higher after MALP-2 application, in association with a bronchial diameter of 0.6-1 mm. The cellular analysis revealed no differences in the number of leukocyte subsets between the control and MALP-2 group. CONCLUSION: MALP-2 is a potent local stimulator and can be used to modulate BALT by repetitive inhalant treatment. The functional significance of enlarged or activated BALT has to be elucidated in future studies. PMID- 12771506 TI - Recruitment of lymphocytes and dendritic cells from the blood to the bronchoalveolar space and the draining lymph nodes after a single intrabronchial application of the lipopeptide MALP-2. AB - OBJECTIVE: It has been shown previously that the synthetic macrophage-activating lipopeptide, MALP-2, is a potent stimulator of the respiratory immune system and an effective adjuvant in the induction of mucosal immune responses. In this study, the migration route of leukocytes from the blood to the bronchoalveolar space and then to the draining lymph nodes was investigated. METHODS: MALP-2 was intratracheally instilled into lungs of Lewis rats. Bronchoalveolar lavage cells as well as cell preparations of other lung compartments such as the marginal vascular pool, the interstitial pool and also the draining lymph nodes were examined 3 days later. RESULTS: The application of MALP-2 induced a pronounced leukocyte accumulation in the bronchoalveolar space and the lung interstitium but not in the marginal vascular pool. A tendency to increased lymphocyte and dendritic cell numbers was observed in the draining lymph nodes. CONCLUSION: Our data indicate the migration of blood cells into the lung interstitium and the bronchoalveolar space in response to MALP-2. Thus, the immune reaction induced by MALP-2 might be of relevance as an adjuvant treatment in inhalant vaccination strategies in the lung. PMID- 12771508 TI - Inflammation versus immunoregulation: what is the key to the development of an effective antiallergy vaccine? AB - The incidence and severity of allergic disorders is currently increasing worldwide. The reason for this development is unclear. However, epidemiological studies and experimental data suggest that this increase may be due to a decline in infectious diseases which induce T helper type 1 (Th1) and/or T regulatory (Tr) responses. Based on these observations, antiallergy vaccines are currently being tested in animal studies aimed at inducing Th1 or Tr responses. This review focuses on novel approaches potentially leading to a vaccine protecting humans from the development of allergic disorders. PMID- 12771509 TI - Animal models of chronic obstructive pulmonary disease: some critical remarks. AB - A variety of animal models of chronic obstructive pulmonary disease (COPD) are available, comprising elastase instillation or inhalation of noxious agents and genetic models like mouse mutants, gene-targeted and transgenic mice. The present review discusses some critical aspects which should be taken into account when evaluating these animal models of human COPD. Critical aspects related to the human disease itself: (1) diagnosis is largely symptom based, and symptoms cannot be mimicked in animals; (2) COPD is not a well-defined entity, but comprises patients with variable contribution of chronic bronchitis, mucus hypersecretion and emphysema, and (3) various factors contribute to the development of COPD, indicating that different pathways may converge into a single endpoint. Critical aspects related to the animals used: (1) species-, strain- and gender-related differences in lung structure and function preclude mimicking some features of the human disease; (2) genetic models frequently present with air space enlargement as a result of the disturbance of early postnatal alveolization (conditionally controlled transgenic animals are recommended); (3) inhalation models frequently use young animals during lung growth, which may preclude distinguishing effects owing to growth retardation or to loss of existing alveolar walls; (4) inhalation of noxious agents may result in reduction of food intake and loss of body weight, which itself may result in emphysema, and (5) the presence of emphysema cannot be concluded based on measurement of air space enlargement alone, but should include the determination of total alveolar surface area. PMID- 12771510 TI - Long-term NO2 exposure induces pulmonary inflammation and progressive development of airflow obstruction in C57BL/6 mice: a mouse model for chronic obstructive pulmonary disease? AB - Chronic obstructive pulmonary disease (COPD) is characterized by chronic inflammation of the airways, development of emphysema and irreversible airway obstruction. Macrophages, neutrophils and CD8+ T cells and their products have been shown to play an important role in the initiation and maintenance of these processes. In contrast, the mechanisms underlying COPD pathogenesis still remain uncertain. This article focuses on the generation of an animal model that mirrors some features of human COPD in association with a progredient airflow limitation. PMID- 12771511 TI - Surfactant proteins SP-A and SP-D as modulators of the allergic inflammation in asthma. AB - Surfactant proteins A and D (SP-A and SP-D) are members of the collectin family and bind to various motifs of microorganisms, particles and allergens. They play an important role in the first-line defense within the lung. Recent research has highlighted that these proteins not only augment innate immune responses to invading microorganisms but also act on adaptive immune functions like dendritic cell maturation and T cell proliferation. Both SP-A and SP-D downregulate the eosinophilic inflammation in murine asthma models and shift the cytokine profile towards a T helper cell type 1 response. In addition, they are effective at alleviating bronchial hyperresponsiveness. Although our knowledge about surfactant proteins as modulators of the allergic inflammatory reaction in asthma is still limited, the idea that surfactant proteins play a role in asthma has attracted increasing attention. In this review, the impact of the lung collectins SP-A and SP-D on asthmatic allergic inflammation and vice versa will be discussed. PMID- 12771512 TI - Transcription factors: new targets for antiallergic therapy. AB - Due to their pivotal role in the fast transmission of signals from cell surface receptors to the nucleus, resulting in gene expression, transcriptional factors play an important part in the physiology of cells. Recently, a number of transcription factors have been identified that are involved in the pathogenesis of allergic diseases. In order to attenuate the development of allergen-mediated diseases, new strategies have been developed to influence these transcription factors in vivo. Among these, the use of oligodeoxynucleotide decoy has emerged as a new tool with promising results for therapeutical and experimental purposes. PMID- 12771513 TI - Development of synthetic pan-selectin antagonists: a new treatment strategy for chronic inflammation in asthma. AB - Asthma is characterized by chronic inflammation of large and small airways maintained by extravasation of leukocytes from the bloodstream into the surrounding peribronchial tissue. The process of extravasation is of crucial importance in inflammation and is mediated by a sequenced and concerted action between different adhesion molecules on endothelial cells and ligands on leukocytes. In this context, initial rolling and tethering is generally considered to be the primary event which is mediated by selectins, a family of glycoproteins comprised of E-, P- and L-selectin. Their role in asthma has been demonstrated in a variety of animal models, showing that all three selectins are involved in the chronic inflammation in asthma. Therefore, selectins are an attractive target where pan-selectin antagonism is the desired treatment strategy. Here, we give an overview of the status of the preclinical and clinical development of bimosiamose, the most advanced synthetic pan-selectin antagonist as a treatment for asthma. PMID- 12771514 TI - Effect of secretin on serum pancreatic enzymes and on the Wirsung duct in chronic nonpathological pancreatic hyperenzymemia. AB - AIM: Chronic nonpathological pancreatic hyperenzymemia (CNPH) is a new syndrome characterized by an increase in serum pancreatic enzymes in the absence of pancreatic disease. The aim of this study was to increase our understanding of this condition by determining the serum pancreatic enzyme response as well as changes in the caliber of the Wirsung duct after secretin stimulation. METHODS: Twenty subjects with CNPH and 9 healthy subjects without CNPH were studied. Blood samples were drawn 5 and 0 min before and 5, 10, 15, 30, 45, and 60 min after intravenous injection of secretin (1 U/kg). Amylase, pancreatic isoamylase, and lipase concentrations were determined. The caliber of the Wirsung duct was measured by ultrasonography. RESULTS: The injection of secretin caused a marked and statistically significant (p < 0.05) increase in serum pancreatic enzymes in the subjects with CNPH that persisted for the duration of the study. The increase over the basal value was in the range of 1.2- to 1.6-fold for amylase, 1.4- to 2.1-fold for pancreatic isoamylase, and 2.6- to 4.2-fold for lipase. In the control subjects the increase was mild, but statistically significant (p < 0.05), ranging from 1.1- to 1.2-fold for amylase, 1.2- to 1.4-fold for pancreatic isoamylase, and 1.5- to 2.2-fold for lipase. The injection of secretin caused a slight increase in the diameter of the Wirsung duct in both groups of subjects, but this was statistically significant only during the first 30 min of the study. CONCLUSIONS: The serum pancreatic enzyme response to secretin was more marked in CNPH than in controls. The Wirsung duct showed no alterations after secretin injection that would help to explain the hyperenzymemia. PMID- 12771515 TI - The bifunctional rat pancreatic secretory trypsin inhibitor/monitor peptide provides protection against premature activation of pancreatic juice. AB - BACKGROUND: In the rat, two forms of the pancreatic secretory trypsin inhibitor, PSTI-I and PSTI-II, are secreted into pancreatic juice. It is assumed that their role is to protect the pancreas from premature activation of the protease-rich pancreatic juice. In the small intestine, PSTI-I, also called 'monitor peptide', is thought to have a different role: PSTI-I competes with protein for activated trypsin. In the presence of a protein-rich meal, free PSTI induces a release of cholecystokinine from the intestine. METHODS: To investigate whether its role as monitor peptide is compatible with the inhibitory, protective function in the pancreas, PSTI-I was chemically synthesized and then renatured. RESULTS: The peptide was almost completely trypsin resistant and exhibited a dose-dependent inhibitory activity to bovine and partially purified rat trypsin. Furthermore, experiments with trypsin- and endopeptidase-activated pancreatic juice demonstrated that its inhibitory capacity was sufficient to prevent premature activation. Binding studies of (125)I-labeled PSTI-I with the putative intestinal receptor using isolated membranes indicated the presence of high-affinity binding sites (k(d) = 5 x 10(-8)M). Binding of PSTI-I could be competed with excess PSTI I or trypsin. In a biological assay system, injections of PSTI-I displayed monitor peptide activity by inducing a dose-dependent trypsinogen release from the pancreas. CONCLUSION: Our experiments support a dual function of PSTI-I: monitoring protein in the gut due to its 'moderate' affinity for trypsin and a protective role in the pancreas. PMID- 12771516 TI - Pancreas cell physiology and pancreatitis cell biology. Summary of a symposium held at the joint meeting of the EPC and the IAP, Heidelberg 2002. PMID- 12771517 TI - Adjuvant chemotherapy in breast cancer: back to the future. PMID- 12771519 TI - Feasibility and morbidity of combined hyperthermia and radiochemotherapy in recurrent rectal cancer--preliminary results. AB - BACKGROUND: The local recurrence rate of colorectal cancer has been significantly reduced due to the use of combined radiochemotherapy. Despite this improvement regarding locally advanced tumour recurrences, the treatment strategy for pre treated patients remains difficult and unresolved. PATIENTS AND METHODS: We analysed treatment and follow-up data of 14 patients with local recurrence of rectal cancer who were treated with radiation therapy (RT), chemotherapy (CT) and regional hyperthermia (RHT) from November 1997 to December 2001. Nine of these patients had received irradiation and CT (= pre-treated patients) in the past. For this group, 30.6-39.6 Gy RT, 5-fluorouracil (5-FU) as a continuous infusion over 5 days per week (350 mg/m(2)/24 h) combined with RHT twice a week was given. The 5 remaining patients (= not pre-treated) received conformal irradiation of 45 Gy with a boost between 9 and 14.4 Gy, combined with continuous infusion of 5-FU on days 1-4, and 29-33 (500 mg/m(2)/ 24 h), and RHT twice a week. Response to therapy was evaluated by means of computed tomography (CT) or magnetic resonance imaging (MRI) and by clinical follow-up. RESULTS: Among 13 evaluated cases, the overall objective response rate was 54% (5 complete responses, 2 partial responses). At mean follow-up of 13.9 months (range 5-32 months) 7 patients were alive. CONCLUSION: The therapeutic regimen appears to be active in the treatment of local recurrences of rectal cancer. Larger-scaled studies are needed to evaluate the potency of hyperthermia in this therapeutic strategy. PMID- 12771518 TI - Randomised trial: One cycle of anthracycline-containing adjuvant chemotherapy compared with six cycles of CMF treatment in node-positive, hormone receptor negative breast cancer patients. AB - AIM: A randomised, controlled clinical trial was initiated in 1984 to test whether 1 cycle of anthracycline-containing adjuvant chemotherapy improves the outcome of breast cancer patients presenting with stage II disease and negative oestrogen and progesterone receptors (ER, PgR), as compared with 6 cycles of dose reduced CMF. PATIENTS AND METHODS: Within 7 years 263 women with stage II breast cancer were randomised either to receive 1 cycle of doxorubicin, vinblastine, cyclophosphamide, methotrexate and 5- fluorouracil (AV-CMF) or to receive 6 cycles of cyclophosphamide, methotrexate and 5-fluorouracil (CMF). Patients were stratified for tumour stage, nodal stage, menopausal status, type of surgery and participating centre. RESULTS: After a median follow-up of 100 months, neither disease-free (DFS) nor overall survival (OS) differed significantly between the two groups. CONCLUSIONS: Compared to 6 cycles of a non-standard low-dose CMF regimen 1 cycle of anthracycline- containing adjuvant chemotherapy failed to improve the outcome in women with stage II receptor-negative breast cancer in terms of DFS and OS. PMID- 12771520 TI - Weekly paclitaxel (Taxol) with or without trastuzumab (Herceptin) in advanced breast cancer: a community-based observation study. AB - BACKGROUND: We evaluated the efficacy and safety of weekly paclitaxel therapy in women with locally advanced or metastatic breast cancer in an unselected hospital based patient population. PATIENTS AND METHODS: Using a specific search tool we were able to find 49 cases in our department-based database MedicoHelp containing more than 36,000 files. All cases were evaluable. The mean number of involved sites was 2.4 (range 1-4) and 38/49 patients had visceral predominant disease. 44 patients received prior antitumor therapies, 33 patients had received prior anthracycline- and/or taxane-containing chemotherapy. Weekly paclitaxel was given at a mean dose of 80 mg/m(2) (day 1, 8, 15, repeated every 28 days), 38 patients with HER2 overexpression had a combination therapy with trastuzumab (Herceptin). Median treatment duration with paclitaxel was 4.4 months (range 0.25-9 months). A dose reduction was necessary in 5 patients. RESULTS: Therapy was tolerated well, with a 12% incidence of hematologic grade 3/4 toxicity and 22% cumulative nonhematologic grade 3 toxicity (asthenia: 5 patients, mucosa/skin: 3 patients, neurotoxicity: 3 patients). Other toxicities were rare. The overall response rate was 63%. 22% of the patients achieved disease stabilization. Symptom improvement was seen in 73% of patients. The mean time to progression was 6.0 months (range 0.25-23 months). CONCLUSIONS: Weekly paclitaxel (mostly in combination with trastuzumab) is a well-tolerated regimen and had a surprisingly high efficacy in this observation study in heavily pre-treated patients. Symptom improvement was seen in the majority of patients who responded to the treatment. PMID- 12771521 TI - Intrapericardial instillation of mitoxantrone in palliative therapy of malignant pericardial effusion. AB - BACKGROUND: Pericardial effusions occurring with pericardial or myocardial metastases often cause serious complications, necessitating temporary or emergency relief by percutaneous pericardiocentesis. However, this often results in recurrences. For long-term therapy success, the intrapericardial instillation of anti-neoplastic agents is an alternative to surgical methods, which are stressful for the patient. Following our positive experiences with mitoxantrone in the treatment of malignant pleural effusions, we applied this substance for the therapy of malignant pericardial effusions. PATIENTS AND METHODS: 16 patients with cytologically verified malignant pericardial effusions (8 with bronchial carcinoma, 7 with carcinoma of the breast, 1 with adenocarcinoma of the stomach) received an intrapericardial instillation of mitoxantrone 1-3 x10-20 mg. Responses were evaluated by echocardiography 30 days after completion of therapy. RESULTS: 12 of 16 patients showed complete remission (no recurrence of a detectable effusion). 3 patients showed a partial remission (recurrence of non drainage-dependent effusion) (CR + PR = 94%). Within the mean follow-up period of 189 days no recurrences occurred. The rate of side effects was low. CONCLUSION: Intrapericardial instillation of mitoxantrone is a feasible and effective palliative method for the control of malignant pericardial effusions with little strain on the patients, short duration of hospital stay, cytotoxic characteristics of the substance with a correspondingly high rate of response and low side effects. PMID- 12771522 TI - Fatigue in the general population. AB - BACKGROUND: Fatigue symptoms are often found in cancer patients. One test to assess fatigue is the Multidimensional Fatigue Inventory (MFI-20). It has been successfully applied to specific groups of cancer patients. However, until now population-based norm values are missing. METHODS: We conducted an investigation on a representative sample of the adult German population, which comprised 2,037 subjects aged 14-92 years. RESULTS: The reliability of the 5 MFI-20 subscales (general fatigue, physical fatigue, reduced activity, reduced motivation, and mental fatigue) is satisfying. The correlations between MFI-20 subscales and the fatigue scale of the quality-of-life questionnaire EORTC indicate convergent validity. As an important result we found that all subscales of the MFI-20 showed a clear and nearly linear dependency on age with higher fatigue values for older subjects (p < 0.0001). Females as compared with males are characterized by higher mean values in all MFI-20 subscales (p < 0.001). CONCLUSIONS: The results show that it is necessary to take into account age and sex when different groups of cancer patients have to be compared. PMID- 12771523 TI - Interferon-a2b reduces neo-microvascular density in the 'normal' urothelium adjacent to the tumor after transurethral resection of superficial bladder carcinoma. AB - BACKGROUND: As angiogenesis represents one of the hallmarks of cancer we investigated whether intravesically administered interferon-a (IFN-a2b) reduces neo-angiogenesis in the 'normal' urothelium adjacent to the tumor in patients with superficial bladder carcinoma after complete transurethral resection (TUR) of the tumor. PATIENTS AND METHODS: In the present study 47 patients after TUR of the tumor were examined. 10 patients (group A) received no further treatment (control group); 37 patients (group B) received intravesical treatment with IFN a2b. The instillations started within 7 days after TUR, were performed weekly for 2 months, twice a month for the next 4 months, and thereafter monthly for 6 more months. Cold cup biopsies were taken before TUR of the transitional cell carcinoma (TCC): from the tumor (T), near tumor (NT) and from normal epithelium (N). Cold cup biopsies 'near tumor', were also taken during follow-up cystoscopy (C1, C2, and C3) 2, 6, and 12 months after TUR, respectively. Angiogenesis was estimated by counting the microvessels detected with CD31 immunostaining. RESULTS: Significant differences of microvascular density (MVD) between patients of group A and B appear after TUR (p < 0.005, Kruskal-Wallis and Wilcoxon test). The MVD difference was maximal 6 months after TUR (C2(A)-C2(B), second cystoscopy) and measured at 12.17 microvessels/ mm(2) (26.2%). CONCLUSION: Our results show that the intravesical administration of IFN-a2b after TUR significantly decreases the angiogenic potential of the 'healthy' urothelium adjacent to the tumor in patients with TCC. This observation could possibly explain, to a certain extent, the mechanism by which IFN-a2b reduces the recurrence rate of primary TCC. PMID- 12771524 TI - Diagnosis of a contralateral second testicular carcinoma by F18-FDG PET. AB - BACKGROUND: In testicular carcinoma the early diagnosis is very important because with an early therapy there are good chances for long-term survival (50-90%). Metastases of a testicular carcinoma are at first lymphogenous, hematogenous only in late stages. CASE REPORT: This is a case report about a 28-year-old man, whose testicular carcinoma (left testis) had already been operated on (unripe teratoma with parts of an embryonic carcinoma, an endodermal sinus tumor and a chorion carcinoma). Because of the elevated tumor marker AFP an FDG PET (F18 fluorodeoxyglucose positron emission tomography) investigation was made. CT (covering thorax, abdomen and pelvis) and ultrasound of the testis showed no pathological results. In the FDG PET a significant pathological FDG uptake in the right testis was found. Histology showed an unripe teratoma with parts of an embryonic carcinoma, an endodermal sinus tumor, and a chorion carcinoma. It was a second carcinoma of the contralateral testis. CONCLUSION: F18-FDG PET was a sensitive and reliable modality for diagnosis in this patient. PMID- 12771525 TI - Treatment of a patient with advanced esophageal cancer with a combination of mitomycin C and capecitabine: activation of the thymidine phosphorylase as active principle? AB - BACKGROUND: Both capecitabine, an oral prodrug of 5-fluorouracil (5-FU), and mitomycin C (MMC) have demonstrated activity as single agents in patients with gastrointestinal cancer. Furthermore, a combination of MMC with infusional 5-FU can induce tumor remission even in patients pretreated with 5-FU. Capecitabine and MMC act synergistically due to an upregulation of the thymidine phosphorylase activity by MMC in a human xenograft model. PATIENT: We sought to exploit these preclinically observed effects in a patient with esophageal cancer who was progressive after a first-line radiochemotherapy with 5-FU and cisplatin. He was treated with a combination of MMC and capecitabine on a compassionate use basis. A rapid remission lasting for about 6 months was observed. CONCLUSION: This is the first report on a combination therapy with capecitabine and MMC. The remission observed in our patient suggests that the preclinically observed synergy has clinical impact. This combination should be further investigated in prospective clinical trials. PMID- 12771526 TI - Adnexal masses in pregnancy. AB - With the widespread use of routine abdominal ultrasound examination during pregnancy, adnexal masses are observed with increasing frequency. Most patients are clinically asymptomatic at the time of presentation, and most of the adnexal masses detected during early pregnancy disappear during the first 16 weeks of pregnancy. Ovarian tumors are estimated to occur in about 1 in 1,000 pregnancies and of these 3% are malignant. Here we present an overview about frequency, diagnostic procedures and pathological characteristics of these ovarian tumors. Moreover, current modalities for treatment during pregnancy are summarized. Surgical treatment of the adnexal masses has to be performed with adequate staging and debulking equal to the treatment of non-pregnant women. However, whereas during organogenesis abortion has to be considered prior to chemotherapy, later in pregnancy surgical debulking as complete as possible, followed by taxol platinum chemotherapy is indicated. If the fetus is not viable at the time of primary surgery, neoadjuvant chemotherapy and complementation of surgery after delivery of the baby should be performed. It should be stressed that chemotherapy for ovarian cancer applied during pregnancy appears to be safe. However, no studies have evaluated the long-term consequences for children exposed to intra uterine chemotherapy. Aspiration of cysts should be avoided, as the correlation between the histological evaluation of an ovarian malignancy and the cytological evaluation of aspirates is poor. Moreover, spillage of malignant cysts is hazardous for the patient. PMID- 12771527 TI - Multidrug resistance in prostate cancer. AB - Advanced hormone-refractory prostate cancer remains a therapeutic challenge, because all available pharmaceutical concepts have been ineffective in improving cancer-specific survival. Failure of chemotherapy may be caused by multidrug resistance (MDR) mechanisms protecting cancer cells against cytotoxic drugs, and the question arises whether prostate cancer is also using MDR principles resulting in resistance against chemotherapeutic agents. In consequence, an array of diverse pathways known to lead to MDR such as MDR1, MRPs, glutathione, and apoptosis have been examined and partially established at varying degrees in hormone-refractory prostate cancer. Thus, evidence keeps accumulating for the involvement of some MDR mechanisms in the chemoresistance of prostate cancer in vitro and in vivo. For some of them, e.g. MRP1, functional expression appears to be probable. This lends credit to the idea that reversal, circumvention, or overcoming of MDR pathways in advanced prostate cancer may be feasible and will lead to new avenues with improved treatment efficacy in otherwise intractable disease. PMID- 12771528 TI - [Home care in Berlin--data on home care of severely ill and dying tumor patients]. PMID- 12771529 TI - Inflammation in uremic patients: what is the link? AB - Uremic patients suffer to an extremely high degree from cardiovascular disease. Cardiovascular disease results mainly from atherosclerotic remodeling of the arterial system. Inflammation is considered to contribute significantly to development of atherosclerosis, and albeit many different factors may lead to inflammation, generation of enhanced oxidative stress is believed to be an important common feature of pro-inflammatory causes. Studies in the general population without renal disease could clearly show that markers of inflammation, in particular C-reactive protein, predict the cardiovascular risk. In this review article, we discuss the presence and the predictive value of inflammation in patients with end-stage renal disease, and analyze whether uremic patients are exposed to specific pro-inflammatory and pro-oxidative conditions. Particular emphasis is set on oxidative stress induced by oxidatively modified lipoproteins and angiotensin II. Based on pathophysiological considerations valid for uremic patients, we discuss therapeutical options that might help to reduce cardiovascular disease in uremic patients. PMID- 12771530 TI - Hypertension and dialysis. AB - The high mortality rate seen in dialysis patients is related to long-standing high blood pressure and the presence of other traditional as well as non traditional risk factors for cardiovascular disease. Hypertension is associated with increased risk for left ventricular hypertrophy, coronary artery disease, congestive heart failure and cerebrovascular complications. High blood pressure is frequent and difficult to control in the dialysis population. Available therapeutic options to normalize blood pressure in these patients include dietary salt and fluid restriction in combination with reduction of dialysate sodium concentration. A possible treatment option for these patients may be long, slow hemodialysis (3 x 8 h per week); short daily hemodialysis (2-3 h 7 times per week); nocturnal hemodialysis (6-7 times overnight per week). Reduction of residual renal function is a major cause of blood pressure increase in the peritoneal dialysis patient population. Therefore, hyperhydration should be avoided. If antihypertensive medication is needed, ACE inhibitors, beta-blockers and/or calcium channel blockers are recommended. Optimal blood pressure in dialysis patients is not different from recommendations for the general population. PMID- 12771531 TI - Rationale for the use of blood volume and temperature control devices during hemodialysis. AB - Despite substantial progress in blood purification techniques over the last three decades, treatment-related hypotensive episodes remain one of the major problems in hemodialysis therapy. There are two main reasons for hypotension occurring during dialysis treatments. First, hypovolemia is frequently induced by rapid fluid removal from the blood compartment which is in excess of refilling of fluids from the interstitial space. Second, many patients fail to support blood pressure by adequate vasoconstriction or increased heart rate as a response to hypovolemia. The capacity to respond adequately to volume contraction may be reduced due to patient- or treatment-related factors, among which heat accumulation within the body plays a major role. Recently, two newer technical developments became commercially available for use in hemodialysis therapy: devices for blood volume and blood temperature control were designed to reduce the incidence of intradialytic hypotension. Although blood volume and temperature measurements are easy to perform today, there is some uncertainty in the dialysis community how and when their use may be helpful and in which patients it is indicated. This review critically discusses the application of blood volume- and temperature-measuring devices with regard to their usefulness in the clinical setting. PMID- 12771532 TI - Treatment options to intensify hemodialysis. AB - At present, three methods are practiced to intensify hemodialysis (HD): 3 times weekly, 8-hour HD, short daily HD and slow daily nocturnal HD. Three times weekly 8-hour dialysis increase both the dialysis dose and time. The longest experiences are in Tassin. Five-year survival in Tassin was better in all age groups compared with the major registries - Japan, EDTA and US Medicare and more obvious for older age groups. The data of Tassin show that increasing the dialysis time provides better blood pressure control, need for no or less antihypertensive drugs, less intradialytic complications, better middle molecule and phosphate clearance, better nutritional status, less requirement for erythropoietin and increased survival. These data of Tassin could be mainly confirmed in our dialysis center. The main difference to Tassin was that in our center most of the patients still need few antihypertensive drugs. The reasons for the difference are that in Tassin the patients are on very low sodium diet (<5 g/day) and in Tassin extracellular volume (ECV) is reduced as far as possible independent of residual renal function. The concept of our center was to preserve residual renal function and accept slightly higher ECV and few antihypertensive drugs. Another concept to intensify HD is short daily HD (6 times/week for 90-180 min). This form of dialysis is offered as in-center and home HD with and without dialysis partner. All studies demonstrated significant improvement of nutritional status, quality of life, control of blood pressure, phosphate and anemia. Survival of AV fistula even with daily double punctures was excellent. The most extensive form of dialysis is slow daily nocturnal dialysis (6 times/week for 8-10 h). This form of dialysis provides excellent urea, phosphate clearance and fourfold increase of beta(2)-microglobulin clearance. Patients discontinue phosphate binders and several patients need phosphate addition to dialysis. Blood pressure control is excellent, all patients are off antihypertensive drugs. Improvement of nutrition, anemia, blood pressure and quality of life is even more pronounced compared to short daily HD or 3 times weekly 8-hour HD. Nocturnal dialysis was able to improve sleep apnea. Nocturnal dialysis is offered only as home HD with and without dialysis partner. Any patient who could be trained for home HD was eligible. Presence of co-morbidities was not a contraindication. PMID- 12771533 TI - Experience with the GENIUS hemodialysis system. AB - The late B. Tersteegen devised a clever way to combine the advantages of a closed tank hemodialysis system with the efficacy and bacteriological safety of a single pass system. The Teerstegen equipment is now marketed as the GENIUS hemodialysis system. For each treatment, fresh dialysis fluid is prepared according to the physician's prescription by mixing sterile ingredients (electrolytes and glucose) with preheated ultrapure water. The total amount of dialysis fluid is put into a thermally insulated glass tank (volume 75 l) of the hemodialysis machine. The filling and emptying process is completely automated. An UV radiator is used for desinfection. Due to a consequent hygienic concept, the system operates with an almost sterile and usually pyrogen-free dialysis fluid. During treatment, fresh dialysis fluid is taken from the top of the system, and the used dialysate is returned to the bottom. There is a sharp interface between the fresh and used dialysis fluids because of a small difference in temperature (1 degrees C). True volumetric ultrafiltration control is simply achieved; ultrafiltration rates between 100 and 1,000 ml/h can be selected as considered appropriate by the physician. MICROBIOLOGICAL EXAMINATION: We cultured more than 2,000 dialysate specimens to examine bacterial contamination, and found either no bacterial growth at all (in the vast majority of the cases) or less than 1 CFU/ml dialysate. CLINICAL EXPERIENCE: We have utilized the GENIUS system since 1994. Meanwhile, more than 40,000 treatments have been performed in our center. Biochemical results of the first 19 stable hemodialysis patients (mean age 66 years, range 45-82), who had been treated with conventional hemodialysis systems for at least 6 months (range 6-157) before changing to GENIUS, were evaluated. We observed an increase of mean serum albumin concentration from 4.1 (+/-0.4 SD) g/dl to 4.8 (+/-0.3) g/dl (p < 0.01) within 6 months. Most patients reported improved well-being. Of these 19 patients, 18 preferred further treatment with the GENIUS system in comparison to conventional hemodialysis machines. CONCLUSION: The GENIUS hemodialysis system permits an individualized therapy of high quality; most patients prefer this system to conventional hemodialysis machines. Serum albumin levels increased significantly from normal to high normal values after change from conventional hemodialysis machines to GENIUS, probably due to less catabolic stress during the hemodialysis sessions. PMID- 12771534 TI - Vitamin C in chronic kidney disease and hemodialysis patients. AB - Increments of oxidative stress have been addressed as one potential cause for the accelerated atherosclerosis of chronic kidney disease patients. Ascorbate represents one of the most prominent antioxidants both in plasma as well as intracellulary, exerting beneficial effects by an inhibition of lipid peroxidation and by reducing endothelial dysfunction. However, in the presence of transition metals like iron, ascorbate may give rise to an increased generation of oxidants, and ascorbylation may impose additional carbonyl stress to uremic patients. Unsupplemented dialysis patients have reportedly lower plasma levels of ascorbate in comparison to healthy controls, mostly due to a loss into the dialysate or, in case of not dialyzed patients, increased urinary losses. Currently, 60 mg of ascorbate are recommended for chronic kidney disease patients, and 1-1.5 g of oral ascorbate/week in case of suspected subclinical ascorbate deficiency or 300 mg parenteral ascorbate/dialysis session, respectively. Ascorbate's role in modifying arterial blood pressure remains unclear, but anemic patients with functional iron deficiency might benefit from short-term, moderately dosed ascorbate supplements. PMID- 12771535 TI - Angiogenin: a novel inhibitor of neutrophil lactoferrin release during extracorporeal circulation. AB - Degranulation of polymorphonuclear leukocytes (PMNL) occurs during extracorporeal circulation. A degranulation-inhibiting protein identical to angiogenin was recently isolated from high-flux dialyzer ultrafiltrate. This protein inhibits the release of lactoferrin and metalloproteinases from PMNL in vitro. In the present study, we investigated end-stage renal disease patients undergoing regular hemodialysis treatment with either high-flux dialyzers (n = 51) or low flux dialyzers (n = 44), and chronically uremic patients undergoing hemodiafiltration (n = 30). Hemodialysis therapy with low-flux polysulfone or cellulose triacetate membranes caused no or only minimal reduction (6 months) with short-term PD treatment, patients with high small solute transport properties (MTAC >11 ml/min, d/p ratio of creatinine >0.72) to patients with low small solute transport and CAPD with APD patients. A peritoneal equilibration test was performed with 1.36% glucose. Dialysate/plasma (D/P) ratio and mass transfer area coefficient (MTAC) of creatinine were calculated and the 4 hour effluent concentration of CA 125 was determined. CA 125 tended to be lower in the long-term PD patients and also in APD patients, but statistical significance was missing. Effluent CA 125 was significantly increased in patients with an MTAC of creatinine >11 ml/min (40.2 +/- 11.2 vs. 20.7 +/- 1.2 U/ml) and in patients with a d/p ratio of creatinine >0.72 (48.2 +/- 11.0 vs. 21.6 +/- 1.6 U/ml). CA 125 and the d/p ratio of creatinine were positively correlated (r = 0.68). The positive correlation of CA 125 with peritoneal small solute transport especially in the early phase of PD treatment indicates an initial correlation of the mesothelial cell mass with the peritoneal surface area. A direct relation between the CA 125 concentration and peritoneal transport is unlikely. In our study the CA 125 effluent concentration tended to be lower in long-term PD patients and also in APD patients, possibly indicating a cell depletory influence of the conventional PD fluid. PMID- 12771539 TI - Cytokine removal in septic patients with continuous venovenous hemofiltration. AB - Despite the progress that has been made in intensive care medicine, multiple organ failure is still associated with high mortality. Apart from the prevention of infectious complications, numerous efforts are being made to improve the treatment of sepsis through adequate antibiotic therapy, the development of new respirator therapies, better control of the hemodynamic situation, and adequate renal replacement therapy. Some authors advocate continuous renal replacement therapy not only for acute renal failure but also for the elimination of inflammatory molecules such as cytokines. Continuous renal replacement therapy improves the cardiovascular hemodynamics in patients with multiple organ failure. Therapeutic options such as volume control, clearance of uremic toxins, correction of acid base disturbances and temperature control are improved. Suitable renal replacement therapy improves not only cardiovascular hemodynamics but also patient survival. In current practice, continuous renal replacement therapy is not used to eliminate mediators such as cytokines. In patients with multiple organ failure and compromised cardiovascular hemodynamics, renal replacement therapy should be carried out as early as possible. In the following review, experimental and clinical findings concerning mediator elimination by continuous and intermittent renal replacement therapy are summarized. PMID- 12771540 TI - How to calculate clearance of highly protein-bound drugs during continuous venovenous hemofiltration demonstrated with flucloxacillin. AB - BACKGROUND: Flucloxacillin is an important antimicrobial drug in the treatment of infections with Staphylococcus aureus and therefore is often used in staphylococcal infections. Furthermore, flucloxacillin has a high protein binding rate as for example ceftriaxone or teicoplanin--drugs which have formerly been characterized as not being dialyzable. METHODS: The pharmacokinetic parameters of 4.0 g flucloxacillin every 8 h were examined in 10 intensive care patients during continuous venovenous hemofiltration (CVVH) using a polyamide capillary hemofilter. In addition, the difficulty of calculating the hemofiltration clearance of a highly protein-bound drug is described. RESULTS: Flucloxacillin serum levels were significantly lowered (56.9 +/- 24.0%) even though only 15% of the drug was detected in the ultrafiltrate. Elimination half-life, total body clearance and sieving coefficient were 4.9 +/- 0.7 h, 117.2 +/- 79.1 ml/min and 0.21 +/- 0.09, respectively. These discrepancies can be explained by the high protein binding of flucloxacillin, the adsorbing property of polyamide and the equation in order to calculate hemofiltration clearance. The unbound fraction of a 4.0 g flucloxacillin dosage facilitates time above the minimum inhibitory concentration (T > MIC) of 60% only for strains up to a minimum inhibitory concentration (MIC) of 0.5 mg/l. CONCLUSION: Based on the data of this study, we conclude that intensive care patients with staphylococcal infections on CVVH should be treated with 4.0 g flucloxacillin every 8 h which was safe and well tolerated. Moreover, further studies with highly protein-bound drugs are recommended to check the classical 'hemodialysis' equation as the standard equation in calculating the CVVH clearance of highly protein-bound drugs. PMID- 12771541 TI - Role of IgE in Th2 cell-mediated allergic airway inflammation. AB - Recent studies with gene knockout mice have demonstrated that T helper 2 (Th2) cell-derived cytokines, including IL-4, IL-5, and IL-13, play important roles in causing allergic airway inflammation. In addition to Th2 cytokines, IgE-dependent activation of mast cells has been suggested to play a role in allergic airway inflammation. In this review, we will discuss the role of IgE in Th2 cell mediated allergic airway inflammation. We used IgE transgenic mice, which enabled us to investigate the role of IgE without the influence of activated T cells and other immunoglobulins. Whereas IgE cross-linking by antigens did not induce eosinophil recruitment into the airways or airway hyperreactivity, IgE cross linking induced CD4+ T cell recruitment into the airways. In addition, when antigen-specific Th2 cells were transferred to IgE transgenic mice, IgE cross linking significantly enhanced antigen-induced eosinophil recruitment into the airways. These findings suggest that IgE-dependent mast cell activation plays an important role in allergic airway inflammation by recruiting Th2 cells into the site of allergic inflammation. PMID- 12771542 TI - The roles of cysteinyl leukotrienes in eosinophilic inflammation of asthmatic airways. AB - Cysteinyl leukotrienes (CysLTs), including LTC(4), LTD(4) and LTE(4), are potent biological mediators generated from arachidonic acid and are produced by inflammatory cells, including eosinophils. Classically, CysLTs have been recognized as powerful spasmogens for bronchial smooth muscle and thus have been implicated in the pathogenesis of asthma. There is increasing evidence that CysLTs also contribute to accumulation of eosinophils within asthmatic airways; CysLTs have been reported to be chemotactic for eosinophils both in vitro and in vivo. CysLTs are also able to enhance the survival of eosinophils. Moreover, LTD(4) augments eosinophil adhesion via beta(2) integrins to intercellular cell adhesion molecule (ICAM)-1, which is constitutively expressed on airway epithelium. Interaction with ICAM-1 enhances the effector functions of eosinophils including the release of specific granule proteins, initiation of the respiratory burst, and generation of additional CysLTs. Thus, CysLTs can augment both accumulation and activation of eosinophils, and may thereby contribute to the phenotypic changes observed in airway eosinophils. We also discuss the effect of CysLT receptor antagonists on airway inflammation in asthma. PMID- 12771543 TI - Th2 immune responses in GATA-3-transgenic mice infected with Heligmosomoides polygyrus. AB - To examine the participation of transcription factor GATA-3 in Th2 immune responses in vivo, we generated transgenic mice having several copies of GATA-3 with LCK promotor. Mice were infected with the intestinal nematode Heligmosomoides polygyrus to induce Th2 immune responses. Upon antigen stimulation, IL-5 and IL-13 production of mesenteric lymph node cells from H. polygyrus-infected mice, was significantly enhanced in GATA-3-transgenic mice compared with nontransgenic control mice. However, IL-4 production was the same in GATA-3-transgenic and control mice. H. polygyrus-infected GATA-3-transgenic mice exhibited significantly more peripheral blood eosinophils and total IgE levels compared with control mice. These results suggest that GATA-3 promotes IL 5 and IL-13 production, and that the function of these cytokines results in eosinophilia and hyper-IgE, respectively. PMID- 12771544 TI - Distinct isoforms of protein kinase C are involved in human eosinophil functions induced by platelet-activating factor. AB - BACKGROUND: Platelet-activating factor (PAF) is a potent stimulator of eosinophils. Recently, treatment with a protein kinase C (PKC) inhibitor which generally inhibits PKC isoforms has been shown to modulate several eosinophil functions in distinct manners, in that PKC inhibition enhanced CD11b expression and cellular adhesion, but inhibited superoxide generation and degranulation in PAF-stimulated human eosinophils. These results suggested that distinct PKC isoforms were likely to be involved in each eosinophil function induced by PAF. We have therefore investigated whether or not the PKC isoforms involved in PAF induced CD11b expression and superoxide generation were different. METHODS: Human eosinophils prepared from healthy volunteers were treated with PKC inhibitors, bis-indolylmaleimide I (BisI; a general PKC inhibitor), myristoylated PKC inhibitor peptide (myr-psiPKC; a PKCalpha, beta and delta inhibitor) and rottlerin (a PKCdelta inhibitor), followed by stimulation with PAF. CD11b expression was determined using flow cytometry and superoxide generation was evaluated using a cytochrome c reduction assay. RESULTS: BisI treatment led to enhancement of PAF-induced CD11b expression, while myr-psiPKC and rottlerin did not. In contrast, PAF-induced superoxide generation was inhibited by treatment with BisI, myr-psiPKC and rottlerin. CONCLUSIONS: PKCalpha, beta and delta are not involved in PAF-induced CD11b expression, but PKCdelta is involved in the PAF induced activation of superoxide anion generation. PMID- 12771545 TI - Interaction with monocytes enhances IL-5 gene transcription in peripheral T cells of asthmatic patients. AB - BACKGROUND: The regulatory mechanisms of IL-5 gene transcription in human peripheral T cells are unclear because the transfection efficiency of plasmid constructs into nontransformed T cells is very low. METHODS: Concanavalin A (ConA)-stimulated blastocytes derived from peripheral blood lymphocytes of asthmatic subjects were transiently transfected with the human IL-5 gene promoter/enhancer-luciferase gene construct, pIL-5 (-511)Luc, and cultured with THP-1 cells (human monocytoid cells) and anti-CD3 monoclonal antibody (mAb). IL-5 level in the culture medium was determined by an enzyme-linked immunosorbent assay. Transcriptional activity of the IL-5 gene was measured by luciferase reporter analysis. RESULTS: ConA-blast lymphocytes of asthmatic patients produced a significant amount of IL-5 upon combined stimulation with anti-CD3 mAb and THP 1 cells, but not with anti-CD3 mAb alone. Costimulation with anti-CD28 mAb also enhanced the anti-CD3 mAb- induced IL-5 production. Accordingly, luciferase activity induced by anti-CD3 mAb stimulation in pIL-5(-511)Luc-transfected ConA blast lymphocytes was increased 1.9- and 3.4-fold by the addition of anti-CD28 mAb and THP-1 cells, respectively. Serial 5' deletion analysis of the reporter gene demonstrated that the cis-regulatory element located at -119 to -80 is critical for anti-CD3 mAb-induced IL-5 gene transcription. CONCLUSIONS: Our present findings provide a useful model reflecting IL-5 gene transcription in human peripheral T cells in vivo, and clearly demonstrate that an interaction with monocytes enhances IL-5 gene transcription. PMID- 12771546 TI - Analysis of gene expression in peripheral blood eosinophils from patients with atopic dermatitis by differential display. AB - To identify the genes related to atopic dermatitis (AD), we compared gene expression in eosinophils from AD patients and healthy volunteers. RNA was prepared from peripheral blood eosinophils. Gene expression was monitored by fluorescent differential display (DD) and real-time RT-PCR. Eighteen new sequences, including expressed sequence tags (ESTs), were expressed at higher levels in eosinophils from AD patients than in those from healthy volunteers. The functions of most of these genes are unknown. We found no correlation between the expression of a particular gene and clinical markers such as the number of eosinophils and the amount of IgE. Multivariate analysis of the gene expression data in each sample showed a very high coefficient of correlation among the copy numbers of each gene. The genes under investigation were also expressed in cultured blood eosinophils after IL-4, IL-5 and IFN-gamma stimulation. We were able to predict the function of some of the sequences by scanning for homologies within either the human or mouse genome databases. The mouse counterpart of one of these genes, intersectin 2, was expressed dramatically, as measured by ear edema, in 1-fluoro-2,4-dinitrobenzene-induced mouse contact dermatitis and in NC/Nga mouse dermatitis. PMID- 12771547 TI - Eosinophil degranulation during pregnancy and after delivery by cesarean section. AB - BACKGROUND: The physiological roles of eosinophils that accumulate in the uterus during pregnancy and of uterus-dwelling mast cells remain unknown. In the present study, we investigated the degranulation of eosinophils and mast cells within the normal course of pregnancy and after delivery by measuring the urinary concentrations of eosinophil-derived neurotoxin (EDN) and N-methylhistamine. METHODS: Spot urine samples from 65 pregnant women and 15 nonpregnant, age matched women were examined. Urinary EDN and N-methylhistamine concentrations were measured by radioimmunoassay and standardized with urinary creatinine concentration. RESULTS: A significant increase in the urinary EDN concentration was observed until the second trimester in the normal pregnancies. The elevated urinary EDN levels decreased after the onset of labor in the third trimester and normalized within 1 month after normal vaginal delivery. In women who underwent a cesarean section, the urinary EDN concentration was significantly higher for up to 1 month after delivery, compared to that in women who underwent a vaginal delivery. In contrast, the urinary N-methylhistamine concentration did not change until the second trimester and was significantly decreased during the third trimester. No significant correlation between the peripheral blood eosinophil count and the urinary EDN concentration was observed in these subjects. CONCLUSIONS: Eosinophils appear to play a role in the progression of pregnancy and recovery after a cesarean section through the degranulation of eosinophils. In addition, mast cell degranulation does not appear to be related to the contraction of uterine smooth muscle during labor. PMID- 12771548 TI - Theophylline attenuates the adhesion of eosinophils to endothelial cells. AB - BACKGROUND: Bronchial asthma is characterized by infiltration of eosinophils and other inflammatory cells into the airways. Binding to adhesion molecules expressed on endothelial cells is an initial step of eosinophil accumulation in the airways of asthmatic patients. Theophylline has been widely used in the treatment of bronchial asthma mainly due to its bronchodilating effect. It has recently been suggested that theophylline induces modulating effects on the survival or functional status of eosinophils. The objective of this study was to determine whether theophylline modifies the adhesive interaction between eosinophils and endothelial cells. METHODS: Eosinophils were isolated from blood of patients with mild asthma. The effects of theophylline on eosinophil adhesion to human umbilical vein endothelial cells (HUVEC) or recombinant adhesion proteins were evaluated. The effect of theophylline on the expression of adhesion molecules on HUVEC was also examined. RESULTS: Theophylline significantly inhibited the eosinophil adhesion induced by formyl-methionyl-leucil phenylalanine (FMLP) or interleukine-5 (IL-5) at a concentration within the therapeutic range. The effect of theophylline on eosinophil adhesion was mimicked by rolipram, a selective phosphodiesterase inhibitor, and N6,2'-O dibutyladenosine 3'5'-cyclic monophosphate (Db-cAMP; cAMP analogue). Finally, theophylline inhibited the expression of ICAM-1 and VCAM-1 on HUVEC stimulated with IL-4 plus TNF-alpha. CONCLUSION: Theophylline showed inhibitory effects on both the adhesive property of eosinophils and the expression of adhesion molecules on endothelial cells, thus suggesting that theophylline attenuates the adhesive interaction between eosinophils and endothelial cells. PMID- 12771549 TI - Single nucleotide polymorphism haplotypes in the cholesteryl-ester transfer protein (CETP) gene and lipid phenotypes. AB - We studied the association between high (HDL) and low-density (LDL) cholesterol concentrations and family-derived haplotypes based on six common SNPs in the cholesteryl-ester transfer protein (CETP) gene. We based our analysis on 201 founders from families recruited throughout Germany. The analysis revealed one subhaplotype block with complete, pairwise, linkage disequilibrium between 5 SNPs located in the promoter and intron 1. The sixth SNP was the well known 1405V polymorphism in exon 14, close to the 3' end of the gene. Four haplotypes accounted for 86% of the entire sample. We found that haplotype associations with HDL, LDL, and the LDL/HDL ratio were more robust than associations with individual SNPs. Moreover, the associations were robust for men, but not for women. Our data suggest an interaction between gender and genetic variation within the CETP gene. PMID- 12771550 TI - A comprehensive method for genome scans. AB - In applications involving the use of genome scans the problem of correcting for multiple testing figures prominently. A frequently used approach is the Bonferroni adjustment, but this is known to be often severely conservative. As an alternative we use the method of importance sampling to accurately and efficiently obtain required exceedance probabilities. This method is comprehensive in the sense that it has application to exceedance probabilities for other classes of test statistics, such as those for linkage disequilibrium or Hardy-Weinberg equilibrium at multiple loci. We illustrate the importance sampling technique by focusing on affected sib pair tests done at a large number of fully informative markers. We demonstrate how our approach can be used to obtain exceedance probabilities for arbitrary marker spacings, and we compare our approach with that of Feingold et al. [1993], which uses the method of large deviations and does not provide the means for adjusting for unequal marker spacing. PMID- 12771551 TI - Entropy as a measure for linkage disequilibrium over multilocus haplotype blocks. AB - OBJECTIVE: The presence of linkage disequilibrium (LD) forms the basis for a range of uses, including the fine-mapping of diseases and studies on human genealogy. Recent findings indicate that single nucleotide polymorphisms (SNP) can occur in blocks of limited haplotypic diversity with high degrees of LD. Commonly used measures for LD, such as r(2) and D', consider only two loci and might miss information to appropriately describe LD in larger haplotypic structures. METHODS: We introduce the Normalized Entropy Difference, epsilon, as a new multilocus measure for LD. A related quantity, deltaS, provides an approximate chi(2) test for the significance of LD. The ability of the measure to detect haplotype blocks is investigated using simulated data sets as well as a real data set previously analyzed by Daly et al. (2001). RESULTS: epsilon allows for arbitrary numbers of loci, describes LD with regard to the loci sequence, and can be interpreted as a multilocus extension of r(2). The application of epsilon to the data sets demonstrated the measure's ability to appropriately describe simultaneous multilocus LD and to detect haplotype blocks. CONCLUSIONS: epsilon is a reasonable multilocus LD measure and might be of potential use in the construction of the human haplotype map. PMID- 12771552 TI - Fine mapping of the schizophrenia susceptibility locus on chromosome 1q22. AB - Schizophrenia is a serious neuropsychiatric illness estimated to affect approximately 1% of the general population. As part of a genome scan for schizophrenia susceptibility loci, we have previously reported a maximum heterogeneity four-point lod score of 6.50 on chromosome 1q21-22 in a group of 22 medium-sized Canadian families, selected for study because multiple relatives were clinically diagnosed with schizophrenia or schizoaffective disorder. We have now conducted fine mapping of this locus in the same set of individuals using 15 genetic markers spanning an approximately 15-cM interval. Parametric linkage analysis with GENEHUNTER v2.1 and VITESSE v2.0 produced a maximum multipoint heterogeneity lod score of 6.50, with a Zmax-1 support interval of <3 cM, corresponding to approximately 1 Mb. Physical mapping and sequence analysis from this region confirmed the presence of an approximately 81-kb tandem duplication, containing low-affinity IgG receptor genes and heat shock protein genes. The sequences of the two copies of this duplication are approximately 97% identical, which has led to the collapse of the two copies into one in the June 2002 NCBI Build 30 of the Human Genome. This duplication may be involved in genomic instability, leading to gene deletion, and so presents an intriguing candidate locus for schizophrenia susceptibility. PMID- 12771553 TI - Confidence intervals for genotype relative risks and allele frequencies from the case parent trio design for candidate-gene studies. AB - OBJECTIVES: Confidence intervals for genotype relative risks, for allele frequencies and for the attributable risk in the case parent trio design for candidate-gene studies are proposed which can be easily calculated from the observed familial genotype frequencies. METHODS: Likelihood theory and the delta method were used to derive point estimates and confidence internals. We used Monte Carlo simulations to show the validity of the formulae for a variety of given modes of inheritance and allele frequencies and illustrated their usefulness by applying them to real data. RESULTS: Generally these formulae were found to be valid for 'sufficiently large' sample sizes. For smaller sample sizes the estimators for genotype relative risks tended to be conservative whereas the estimator for attributable risk was found to be anti-conservative for moderate to high allele frequencies. CONCLUSIONS: Since the proposed formulae provide quantitative information on the individual and epidemiological relevance of a genetic variant they might be a useful addition to the traditional statistical significance level of TDT results. PMID- 12771554 TI - Gastric cancer prevalence in patients with liver cirrhosis. AB - An increased risk for gastric cancer in patients with liver cirrhosis has recently been reported in epidemiological studies. The present endoscopic study was performed to further evaluate whether people with cirrhosis are at increased risk for gastric cancer development. We reviewed the medical records of all cirrhotic patients referred to our Endoscopic Service for portal hypertension screening and, therefore, cases of latent gastric cancer were observed. For a comparison, the prevalence (age and sex standardized) of latent gastric cancer in the general population was estimated hypothesizing a latency period of 5 years. Overall, 1379 patients with cirrhosis were selected from a total of 15 791 endoscopically examined different patients observed during the period 1982-1997. Histological assessment revealed the presence of gastric cancer in 10 patients (9 males and 1 female). There was a significant 2.6-fold (P<0.01) increase in prevalence of gastric cancer compared with that expected in our cirrhotic patients. In conclusion, our findings confirm that liver cirrhosis would seem to be a risk factor for the development of gastric cancer. Other studies are needed to evaluate the pathogenic mechanisms involved. PMID- 12771555 TI - Gastric cancer and other organ cancer history in the parents of patients with gastric cancer. AB - A case-control study in a population from the Eastern Black Sea region of Turkey was performed to evaluate the risk of gastric cancer development in patients with gastric cancer reporting gastric cancer or other organ cancer history in their parents. Gastric cancer and/or other organ cancer history in the parents were found in 215 of 1240 patients with gastric cancer versus 73 of 1240 controls (odds ratio (OR) 3.35, P<0.001). The frequency of gastric cancer history among the parents was significantly different between the patients and the controls (148 versus 25, respectively; OR 6.59, P<0.001). The frequency of other organ cancer history was not significantly different between the patients and the controls (63 versus 48, respectively; OR 1.33, P>0.05). The frequency of gastric cancer history was significantly higher than the frequency of other organ cancer history in the parents of the patients (OR 4.51, P<0.001). Gastric cancer history was significantly higher in first- to third-degree relatives of the patients reporting gastric cancer and/or other organ cancer history in their parents than in the controls (OR 14.72, P<0.001). Familial clustering of gastric cancer defined by the presence of at least four cancer cases in family members, including parents, was reported by 12% of the patients. Overall, the result of this study of gastric cancer and other organ cancer history in the parents of patients with gastric cancer may suggest a genetic susceptibility for gastric carcinoma. A high risk of gastric cancer occurred in subjects reporting a gastric cancer history in their parents, and there was a higher predisposition to gastric cancer compared with other organ cancer in relatives and a familial clustering of the disease. PMID- 12771556 TI - Malignant carcinoids in the USA, SEER 1992-1999. An epidemiological study with 6830 cases. AB - Most of our knowledge on the epidemiology of carcinoid tumours in the USA relies on studies updated during the 1980s. The aim of this study was to describe USA population-based case-series incidence in the period 1992-1999. Data were retrieved from a Surveillance Epidemiology and End Results (SEER) Program public use file. Incidence rates, male-to-female standardized ratios, estimated annual percentage changes and 5-year relative survival rates were computed, and 6830 malignant carcinoid incident during 1992-1999 were analysed. The overall age adjusted incidence rates were 1.9 per 100 000 for men and women. There was an excess in women for lung and stomach carcinoids, while rates were higher among men for carcinoids of the small intestine and rectum. From 1992 to 1999, incidence rates showed a 3% estimated annual increase. Thirteen per cent of the cases had metastasis at diagnosis. In the present series, 24% of carcinoid patients had more than one tumour. The overall 5-year relative survival rate was 82%. In conclusion, improvements in diagnostic techniques seem to have influenced the epidemiology of carcinoid tumours, leading to a slightly different site distribution and a better stage distribution at diagnosis. Increasing incidence and improving survival rates were shown. PMID- 12771557 TI - Tomato and garlic can modulate azoxymethane-induced colon carcinogenesis in rats. AB - Tomato (Lycopersicon esculentum) and garlic (Allium cepa) are important constituents of the human diet. Compounds like diallyl sulfides, diallyl disulfides and quercetin, which are active components of garlic, have known anti inflammatory, antimutagenic activities. Similarly, active components in tomato, such as kaempferol and chlorogenic acid, have antimutagenic activities and lycopene is the most active oxygen quencher with potential chemopreventive activities. In view of this, an endeavour was made to evaluate the anticarcinogenic effect, if any, of tomato and garlic consumption individually and in combination on azoxymethane-induced colonic precancerous lesion, the aberrant crypt foci in animal model. Sprague-Dawley rats (4-5 weeks old) were injected with azoxymethane (15 mg/kg b.w.) and orally administered with 2% (w/v) of tomato, garlic and a combination of both. After 12 weeks of first azoxymethane injection, colons were assessed for aberrant crypt foci and compared with the carcinogen control group. Lipid peroxidation level and glutathione-S-transferase (GST) activity were assessed in liver as well as in colon. Furthermore, in situ cell proliferation and apoptosis were estimated using the Brdu incorporation method and TUNEL method respectively. It was observed that aberrant crypt foci were reduced in all treated groups (by 32.11% in garlic, by 76.14% in tomato and by 55.96% in the combination group). Among treated groups, GST activity was found to be induced in both liver and colon, whereas considerable reduction in lipid peroxidation level was observed in liver as well as in colon with respect to the carcinogen control group. Significant reduction in Brdu labelling index and increase in apoptotic index in colon was noted in the treated groups. These results suggest that tomato and garlic suspensions have a protective effect on colon carcinogenesis, which is mediated by modulation of different biological pathways during carcinogenesis. PMID- 12771558 TI - Practice of opportunistic PSA screening in the Florence District. AB - The widespread use of the prostate-specific antigen (PSA) test had a great impact on the rise of incidence of prostate cancer. The magnitude of opportunistic screening by PSA varies across countries, being highest in the US, and considerably lower in Europe. To estimate the opportunistic use of PSA over the period of one year (2000) in the District of Florence, we analysed the Regional Database of diagnostic exams. According to the Regional Database, subjects having at least one PSA test during the year 2000 ranged from 5.8 to 6.7%, 8.8-10.8%, 11.7-15.0%, 16.5-22.3%, 18.0-24.3%, 17.0-23.8% and 14.1-18.3% in the 50-54, 55 59, 60-64, 65-69, 70-74, 75-79 and >79 years age groups, respectively. Minimum and maximum values are reported according to exclusion or inclusion of PSA determinations with incomplete subject identification data. Such a high use of PSA should deserve special attention from both the medical and health care provider community. The aim of the present study was to evaluate the frequency of opportunistic screening by PSA in the District of Florence in the general population. PMID- 12771559 TI - Association between CYP2E1 polymorphisms and susceptibility to prostate cancer. AB - Several genetic alterations have been associated with sporadic prostate cancer (PCa). In this study, the association between RsaI and DraI polymorphisms of CYP2E1 and PCa risk was analysed in a case-control study of 227 individuals using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Regarding DraI polymorphisms, the DD genotype is over-represented in PCa cases when compared with the control group (odds ratio (OR) 2.12; 95% confidence interval (CI) 1.11-4.05; P=0.022). Regarding the RsaI polymorphism, no significant differences were found. The results of this study indicate that DraI polymorphisms of the CYP2E1 gene may be associated with a twofold increased risk for the development of PCa. PMID- 12771560 TI - Reporting of performance indicators of mammography screening in Europe. AB - We compared short-term indicators for service mammography screening in Europe. Data were available from 17 programmes, although not all programmes provided a comprehensive reporting. More than 90% of the target population had been screened within the last 3 years in the WE trial, whereas only two-thirds of women in England and Copenhagen had been screened within the last 3 years, which will delay or reduce the effect of screening compared with the trial. Participation was highest in sparsely populated areas. Detection rates at first screen reached three times the baseline in Copenhagen, the Netherlands and North-West England. The clinical characteristics of screen-detected cases were badly reported. Given their importance for the long-term effect of screening, further data are warranted. Sensitivity and specificity could be measured only indirectly; they showed, however, considerable variation between programmes. Fyn, Florence and Stockholm had succeeded in combining high specificity with high sensitivity. With different recall policies, different proportions of women will experience a false positive test; expected numbers after three screens were 14%, 10% and 1%, for England, Copenhagen and the Netherlands, respectively. Based on the observed wide variation in short-term indicators, a similar wide variation is expected in the effect of screening on breast cancer mortality. PMID- 12771561 TI - Performance indicators of organized cervical screening in Romagna (Italy). AB - The availability of published data from organized cervical screening programmes in southern Europe is scant. In the Italian area of Romagna, a first round of organized screening (based on a 3-yearly Pap smear for women aged 25-64 years) was initiated between December 1995 and January 1997 and was completed in an average of 42 months (range 36-48 months). The target population included 305 478 women. Of these, 253 949 were eligible and received a personal letter of invitation. Age-specific screening performance indicators were calculated according to standard methods. The response rate within 6 months of invitation was 49.1% (n=124 621). The total participation rate including women who presented later was 61.7% (n=156 735). The recall rate was 35.2 per 1000 of participants (n=5514). Positive cytology results were distributed as follows: atypical squamous cells of un-determined significance/atypical glandular cells of undetermined significance (ASCUS/AGUS) 40.1%, low-grade squamous intraepithelial neoplasia (LGSIL) 48.6%, high-grade squamous intraepithelial neoplasia (HGSIL) 10.7% and carcinoma 0.7%. Compliance to colposcopy follow-up was 93.4% (n=5149). The biopsy rate was 52.4% (n=2696) of patients undergoing colposcopy. The detection rate was 4.5 per 1000 of participants (n=707) for CIN2-3 and 0.5 (n=75) for invasive carcinoma. The proportion of microinvasive carcinomas was 36.0% (n=27). The positive predictive value for CIN2-3/carcinoma was 5.8% for the cytology reports of ASCUS/AGUS, 7.6% for those of LGSIL, 76.5% for those of HGSIL, and 100.0% for those of carcinoma (80.4% for combined HGSIL/carcinoma). The ratio of observed to expected (or prevalent to incident) cases of invasive carcinoma was 2.35 (95% confidence interval (CI) 1.85-2.95). In conclusion, most early results of the programme were compatible with an acceptable performance. PMID- 12771562 TI - Dietary and lifestyle factors in relation to plasma insulin-like growth factor I in a general population sample. AB - There is evidence that the insulin-like growth factor system (IGF), particularly IGF-I, is important in human carcinogenesis. We studied in a general, though not strictly random population sample of 620 adults, the relationship of IGF-I to demographic, lifestyle and dietary factors, the latter ascertained through an extensive validated questionnaire. Plasma IGF-I levels declined significantly with age and the decline was more evident among women than among men. Tobacco smoking, body mass index and regular physical activity were unrelated to this hormone and a positive association with height was not statistically significant. Neither protein nor carbohydrate intake was related to plasma IGF-I levels but there was inconsistent evidence that ethanol intake may be inversely associated with plasma IGF-I and saturated and polyunsaturated lipids may be positively associated with it. The findings are evaluated in conjunction with evidence indicating that the incidence of cancer is lower among women than among men, height is a risk factor for several forms of cancer, and saturated and polyunsaturated lipids have been more closely linked to human and animal carcinogenesis than monounsaturated lipids. PMID- 12771563 TI - Tobacco-smoking prevalence among physicians and nurses in countries with different tobacco-control activities. AB - The aim of the study was to compare rates of smokers among physicians and nurses in the USA, a country with relatively high levels of activity in tobacco control, with those in a country with low levels of tobacco-control efforts. Analysis of interview data in three cross-sectional population studies was carried out. The tobacco-smoking rate of the physicians in the country with low prevention activity dropped to 18%, which is still much higher than the smoking rate in the US and other European countries. In conclusion, prevention activity on a national level might contribute to reducing the rate of current smokers among physicians to a large extent, less so in nurses. PMID- 12771564 TI - A regular contribution from Lars Ovesen, which we hope will help the busy scientist in keeping up with the literature Fine Arts. PMID- 12771565 TI - Increased risk of breast cancer in relatives of malignant melanoma patients from families with strong cancer familial aggregation. AB - The aim of this study was to evaluate the risk of occurrence of malignancies of different site of origin in patients with malignant melanoma (MM) of the skin and their first-degree relatives from families with cancer familial aggregations with unknown pathogenetic background (CFA). We analysed tumour spectrum and age at diagnosis of malignancies in 51 families with MM/CFA. In addition, we evaluated observed frequency (OF); expected frequency (EF); and relative risk (RR) of occurrence of malignancies in these families. In all cases peripheral blood examination of common Polish founder BRCA1 mutations was performed. In 25 families, we analysed loss of heterozygosity of BRCA1 and BRCA2 genes. We identified two subgroups of cases: 22 MM/CFA families with MM diagnosed before 55 years (< or =55 MM/CFA) and 29 MM/CFA families with MM diagnosed after 55 (>55 MM/CFA). In these families we observed increased proportion of breast cancers: 17.52% in the first subgroup (mean age of diagnosis 48.5) and 12.15% in the second subgroup. The odds ratio for breast tumours occurring before 50 in < or =55 MM/CFA families was 3.71. We also observed increased numbers of liver cancers, CSU and leukaemias. OF and EF analyses revealed increased risk of occurrence of cancers of breast (OF 10.4%, EF 4.5%) and liver (OF 1.9%, EF 0.8%) in women from MM/CFA families, RR for breast tumours was approximately 3.3 in < or =55 MM/CFA families. Molecular examination of MM/CFA families revealed no alterations within the BRCA2 gene and one germline mutation of the BRCA1 gene. In conclusion, it seems to be justified to consider systematic breast surveillance beginning at the age around 35-40 years as an option in women from < or =55 MM/CFA families. PMID- 12771566 TI - Effects of hypothermia on energy metabolism in Mammalian central nervous system. AB - This review analyzes, in some depth, results of studies on the effect of lowered temperatures on cerebral energy metabolism in animals under normal conditions and in some selected pathologic situations. In sedated and paralyzed mammals, acute uncomplicated 0.5- to 3-h hypothermia decreases the global cerebral metabolic rate for glucose (CMR(glc)) and oxygen (CMRo(2)) but maintains a slightly better energy level, which indicates that ATP breakdown is reduced more than its synthesis. Intracellular alkalinization stimulates glycolysis and independently enhances energy generation. Lowering of temperature during hypoxia-ischemia slows the rate of glucose, phosphocreatine, and ATP breakdown and lactate and inorganic phosphate formation, and improves recovery of energetic parameters during reperfusion. Mild hypothermia of 12 to 24-h duration after normothermic hypoxic ischemic insults seems to prevent or ameliorate secondary failures in energy parameters. The authors conclude that lowered head temperatures help to protect and maintain normal CNS function by preserving brain ATP supply and level. Hypothermia may thus prove a promising avenue in the treatment of stroke and trauma and, in particular, of perinatal brain injury. PMID- 12771567 TI - No role for interleukin-18 in acute murine stroke-induced brain injury. AB - There is now extensive evidence to show that the cytokine interleukin-1 (IL-1) contributes directly to reversible and permanent ischemic brain damage in rodents. Because interleukin-18 (IL-18) shares many structural and functional similarities with IL-1, the authors tested the hypothesis that IL-18 contributes directly to ischemic brain damage in mice exposed to focal, reversible (15-minute or 30-minute) middle cerebral artery occlusion. IL-18 expression was not induced acutely by middle cerebral artery occlusion, and deletion of the IL-18 gene (IL 18 knockout mice) did not affect infarct volume. The present results suggest that IL-18 does not contribute to acute ischemic brain damage. PMID- 12771568 TI - Relation between cerebral blood flow and metabolism explained by a model of oxygen exchange. AB - The cerebral blood flow (CBF) and cerebral metabolic rate of oxygen (CMRo(2)) are major determinants of the contrast in functional magnetic resonance imaging and optical imaging. However, the coupling between CBF and CMRo(2) during cerebral activation remains controversial. Whereas most of the previous models tend to show a nonlinear coupling, experimental studies have led to conflicting conclusions. A physiologic model was developed of oxygen transport through the blood-brain barrier (BBB) for dynamic and stationary states. Common model simplifications are proposed and their implications for the CBF/CMRo(2) relation are studied. The tissue oxygen pool, the BBB permeability, and the hemoglobin dissociation curve are physiologic parameters directly involved in the CBF/CMRo(2) relation. We have been shown that the hypothesis of a negligible tissue oxygen pool, which was admitted by most of the previous models, implies a tight coupling between CBF and CMRo(2). By relaxing this hypothesis, a real uncoupling was allowed that gives a more coherent view of the CBF/CMRo(2) relation, in better agreement with the hypercapnia data and with the variability reported in experimental works for the relative changes of those two variables. This also allows a temporal mismatch between CBF and CMRo(2), which influences the temporal shape of oxygenation at the capillary end. PMID- 12771569 TI - The hemodynamic impulse response to a single neural event. AB - This article investigates the relation between stimulus-evoked neural activity and cerebral hemodynamics. Specifically, the hypothesis is tested that hemodynamic responses can be modeled as a linear convolution of experimentally obtained measures of neural activity with a suitable hemodynamic impulse response function. To obtain a range of neural and hemodynamic responses, rat whisker pad was stimulated using brief (or=2 days of duration, the longest being 22.8 days. Thirty-one of 40 patients (77.5%) required intubation and mechanical ventilation during 36 of the 57 admissions, and 15 of these episodes (41.6%) ended with the patient's discharge from the ICU. Of ten patients with respiratory failure associated with pulmonary infection, there were no survivors among those who remained ventilated at 48 hrs (n = 8). Four patients who required mechanical ventilation (12.9%) were alive at the 6 month follow-up. The majority of patients who died in the ICU did so after either withdrawal (65%) or limitation (22%) of treatment. CONCLUSIONS: Despite the generally poor prognosis for pediatric patients admitted to the ICU after BMT, intensive care continues to play an important role in the care of these patients. Although it is clear that patients who require mechanical ventilation have a worse prognosis, we were unable to identify factors that accurately predict with 100% sensitivity which patients will not survive. Those patients requiring mechanical ventilation due to pneumonitis have a particularly poor outcome, and our findings support the limitation of intensive care in certain circumstances. Decisions regarding treatment options and limitation of care in this group of patients should be based on ongoing outcome research in this field. PMID- 12771595 TI - Mechanical effects of airway humidification devices in difficult to wean patients. AB - OBJECTIVE: To evaluate the influence of airway humidification devices on the efficacy of ventilation in difficult to wean patients. DESIGN: A prospective, randomized, controlled physiologic study. SETTING: A 22-bed medical intensive care unit in a university hospital. PATIENTS: Chronic respiratory failure patients. INTERVENTIONS: Performances of a heated humidifier and a heat and moisture exchanger were evaluated on diaphragmatic muscle activity, breathing pattern, gas exchange, and respiratory comfort during weaning from mechanical ventilation by using pressure support ventilation. Eleven patients with chronic respiratory failure were submitted to four pressure support ventilation sequences by using the heated humidifier and the heat and moisture exchanger at two different levels of pressure support ventilation (7 and 15 cm H(2)O). MEASUREMENT AND MAIN RESULTS: Compared with the heated humidifier and regardless of the pressure support ventilation level used, the heat and moisture exchanger significantly increased all of the inspiratory effort variables (inspiratory work of breathing expressed in J/L and J/min, pressure time product, changes in esophageal pressure, and transdiaphragmatic pressure; p <.05) and dynamic intrinsic positive end-expiratory pressure (p <.05). Similarly, the heat and moisture exchanger produced a significant increase in Paco(2) (p <.01) responsible for severe respiratory acidosis (p <.05), which was insufficiently compensated for despite a significant increase in minute ventilation (p <.05). This resulted in respiratory discomfort for all patients with the heat and moisture exchanger (p <.01). Adverse effects were partially counterbalanced by increasing the pressure support ventilation level with the heat and moisture exchanger by >or=8 cm H(2)O. CONCLUSIONS: The type of airway humidification device used may negatively influence the mechanical efficacy of ventilation and, unless the pressure support ventilation level is considerably increased, the use of a heat and moisture exchanger should not be recommended in difficult or potentially difficult to wean patients with chronic respiratory failure. PMID- 12771596 TI - Outcome and attributable cost of ventilator-associated pneumonia among intensive care unit patients in a suburban medical center. AB - OBJECTIVE: To determine the attributable cost of ventilator-associated pneumonia from a hospital-based cost perspective, after adjusting for potential confounders. DESIGN: Patients admitted between January 19, 1998, and December 31, 1999, were followed prospectively for the occurrence of ventilator-associated pneumonia. Hospital costs were defined by using the hospital cost accounting database. SETTING: The medical and surgical intensive care units at a suburban, tertiary care hospital. PATIENTS: Patients requiring >24 hrs of mechanical ventilation. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: We measured occurrence of ventilator-associated pneumonia, in-hospital mortality rate, total intensive care unit (ICU) and hospital lengths of stay (LOS), and total hospital cost per patient. Ventilator-associated pneumonia occurred in 127 of 819 patients (15.5%). Compared with uninfected, ventilated patients, patients with ventilator associated pneumonia had a higher Acute Physiology and Chronic Health Evaluation II score on admission (p <.001) and were more likely to require multiple intubations (p <.001), hemodialysis (p <.001), tracheostomy (p <.001), central venous catheters (p <.001), and corticosteroids (p <.001). Patients with ventilator-associated pneumonia were more likely to be bacteremic during their ICU stay (36 [28%] vs. 22 [3%]; p <.001). Patients with ventilator-associated pneumonia had significantly higher unadjusted ICU LOS (26 vs. 4 days; p <.001), hospital LOS (38 vs. 13 days; p <.001), mortality rate (64 [50%] vs. 237 [34%]; p <.001), and hospital costs (70,568 dollars vs. 21,620 dollars, p <.001). Multiple linear regression, controlling for other factors that may affect costs, estimated the attributable cost of ventilator-associated pneumonia to be 11,897 dollars (95% confidence interval = 5,265 dollars-26,214 dollars; p <.001). CONCLUSIONS: Patients with ventilator-associated pneumonia had significantly longer ICU and hospital LOS, with higher crude hospital cost and mortality rate compared with uninfected patients. After we adjusted for underlying severity of illness, the attributable cost of ventilator-associated pneumonia was approximately 11,897 dollars. PMID- 12771597 TI - Antiseptic chamber-containing hub reduces central venous catheter-related infection: a prospective, randomized study. AB - OBJECTIVE: The hub connecting the catheter and the infusion equipment is a common portal of entry for bacteria causing catheter-related sepsis. We assessed the efficacy of a new hub model (Segur-Lock) that incorporates an antiseptic chamber filled with 3% iodinated alcohol in preventing endoluminal catheter contamination and catheter-related bloodstream infection arising at the hub. DESIGN: Prospective, randomized, multicenter study. SETTING: Seven medical and surgical adult intensive care units in Spain. PATIENTS: A total of 230 patients aged 18 yrs or older requiring the insertion of a nontunneled central venous catheter for >or=6 days from January 1, 1998, to April 30, 1999. INTERVENTIONS: Patients were randomized at the time of catheter insertion to receive catheters with standard Luer-lock connector (control group, n = 114) or catheters with the new hub model (n = 116). MEASUREMENTS AND MAIN RESULTS: Skin, catheter tip, and hub cultures were performed at the time the catheter was withdrawn because therapy was terminated or due to suspicion of sepsis, in which case peripheral blood and infusate cultures were simultaneously taken. Catheter-related bloodstream infection was diagnosed in 19 (8.3%) patients. Catheters were more often withdrawn because of suspicion of infection in the control group (43.8%) than in the new hub model group (30.1%, p <.035). The prevalence of culture-positive catheter hubs without associated bacteremia (colonization) was higher in the control group (14.4% vs. 4.3%, p <.001). Catheter-related bloodstream infection from the catheter hub also occurred more frequently in controls than in patients assigned to the new hub model (7% vs. 1.7%; p <.049; relative risk, 4.14; 95% confidence interval, 0.8-19). CONCLUSIONS: This new antiseptic chamber-containing hub has proved to be effective in preventing endoluminal bacterial colonization and catheter-related bloodstream infection from hub contamination in intensive care unit patients with central venous catheters inserted for >or=6 days. PMID- 12771598 TI - Acute respiratory distress syndrome after kidney transplantation: epidemiology, risk factors, and outcomes. AB - OBJECTIVE: To determine the rate of the acute respiratory distress syndrome (ARDS) after kidney transplantation and to identify risk factors associated with the development of ARDS after kidney transplantation and outcomes for patients diagnosed with ARDS in this setting. DESIGN: Retrospective analysis of the national registry for end-stage renal disease in the United States. PATIENTS: We studied all patients who underwent kidney transplantation between July 1, 1994 and June 30, 1998 and identified patients diagnosed with ARDS. The diagnosis of ARDS was based on coding of patients records. We also compared the rate of ARDS after kidney transplantation with the rate of ARDS in the remainder of the U.S. population based on the results of the National Hospital Discharge Survey for 1997. MEASUREMENTS AND MAIN RESULTS: During the study period, 42,190 kidney transplantations were performed in the United States and ARDS was diagnosed in 86 of these subjects (0.2%) resulting in an annualized rate of ARDS of 51.0 cases per 100,000 patients per year. The rate of ARDS after kidney transplantation was significantly higher than the reported rate of ARDS in the U.S. population (p <.050). Demographic factors, indications for transplantation, comorbid illness, antigen mismatch, cytomegalovirus status, and development of rejection did not correlate with the development of ARDS. Of the immunosuppressive agents (e.g., cyclosporine, FK-506, mycophenolate mofetil, azathioprine, OKT-3, antilymphocyte globulin), only the use of antilymphocyte globulin when used to treat rejection was linked with an increased risk for ARDS (odds ratio: 3.85; 95% confidence interval: 1.36 to 10.87). Subjects with graft failure were 2.70 (95% confidence interval: 1.33 to 5.52) times more likely to develop ARDS. The 28-day mortality in subjects with ARDS was 52.1%. The 3-yr survival after kidney transplantation was 88.9% in those without ARDS compared with 57.8% in persons with ARDS (p <.001). CONCLUSIONS: Although ARDS is a rare event after kidney transplantation, undergoing renal transplantation increases the risk for ARDS. Among patients receiving kidney transplants, graft failure and the use of antilymphocyte globulin for rejection are associated with the development of ARDS. Patients who develop ARDS after kidney transplantation face significant mortality. PMID- 12771599 TI - Endogenous glycosides in critically ill patients. AB - OBJECTIVE: To determine the incidence of critically ill patients displaying endogenous digitalis-like-immunoreactive substances (DLIS) and to examine the relationship of these hormones to routine laboratory variables, the underlying disease, myocardial function, hemodynamic status, severity of illness, systemic inflammation, and mortality rate. DESIGN: Sera of 401 consecutive critically ill patients, not treated with cardiac glycosides, were analyzed for DLIS (digitoxin and digoxin, TDx; Abbott Diagnostics, North Chicago, IL) and endogenous ouabain. Normal values of endogenous ouabain were determined in 62 healthy volunteers. We measured pro- and anti-inflammatory mediators (L-selectin, tumor necrosis factor alpha, interleukin-1beta, interleukin-2, interleukin-6, interleukin-10), C reactive protein, and serum amyloid A protein as well as patients' Acute Physiology and Chronic Health Evaluation II and Goris scores. In a subgroup of patients with a pulmonary artery catheter (n = 95), we determined cardiac output, pulmonary artery occlusion pressure, systemic and pulmonary vascular resistance, left ventricular stroke volume, and right and left stroke work. SETTING: Two surgical intensive care units of an university hospital. SUBJECTS: Sera of 401 consecutive critically ill patients. INTERVENTIONS: Blood sampling. MEASUREMENTS AND MAIN RESULTS: Of the 401 patients tested, 343 had nonmeasurable DLIS concentrations (DLIS-negative), and 58 (14.5%) had positive digoxin (n = 18) or digitoxin (n = 34) concentrations (DLIS-positive) or were positive in both tests (n = 6). Mean endogenous ouabain concentrations were nine-fold increased in DLIS positive (3.59 +/- 1.43 nmol/L) and three-fold increased in DLIS-negative (1.34 +/-.81 nmol/L) patients compared with controls (0.38 +/- 0.31 nmol/L). DLIS and ouabain concentrations closely correlated with the Acute Physiology and Chronic Health Evaluation II and Goris score and were associated with increased concentrations of transaminases, bilirubin, aldosterone, cortisol, serum creatinine, fractional sodium excretion, proinflammatory mediators, C-reactive protein, and serum amyloid A (p 100 pg/mL had a significantly lower nadir ERMBT compared with subjects with a peak interleukin-6 of <100 pg/mL (35.5% +/- 5.2% vs. 74.7% +/- 5.1%, p <.001). CONCLUSIONS: Acute inflammation after elective surgery was associated with a significant decline in cytochrome P450 3A4 activity, which is predictive of clinically important changes in the metabolism of commonly used drugs that are substrates for this enzyme. PMID- 12771601 TI - Bioavailability of gatifloxacin by gastric tube administration with and without concomitant enteral feeding in critically ill patients. AB - OBJECTIVE: Sequential intravenous-to-oral antimicrobial therapy with highly bioavailable antiinfective agents such as the fluoroquinolones may improve patient safety and decrease cost of infection management. However, physiologic changes associated with critical illness may alter drug absorption, distribution, and clearance, and concomitant enteral feeding may decrease fluoroquinolone bioavailability. We evaluated the effect of critical illness and concomitant gastric tube feeding on gatifloxacin bioavailability. DESIGN: Prospective, randomized, single-dose, two-way crossover, pharmacokinetic study. SETTINGA tertiary, level-one, trauma center. PATIENTS: Sixteen critically ill patients (baseline Acute Physiology and Chronic Health Evaluation II score >or=16) tolerating enteral nutrition administered by gastric tube (NG) for >or=12 hrs were randomized to receive gatifloxacin concurrently with continuous tube feeding or with interrupted tube feeds. Patients with renal insufficiency or those receiving concomitant fluoroquinolone therapy or postpyloric feeding were excluded. Patients received gatifloxacin 400 mg either by the intravenous or NG route followed by the alternative dosage form after a 72-hr washout period. MEASUREMENTS AND MAIN RESULTS: Serial serum gatifloxacin concentrations (from 5 mins to 24 hrs) were analyzed using a validated high-performance liquid chromatography method. Bioavailability was determined as the ratio of NG/intravenous area under the concentration-time curve (AUC infinity ) measured by the trapezoidal method. Although there was no difference in the bioavailability between NG (AUC infinity : 38.0 [range 20.1 to 48.5] microg x h/mL) and intravenous (AUC infinity : 39.5 [range 24.1 to 63.1] microg x h/mL, p =.60) gatifloxacin (bioavailability: 98.5% [range 61.1% to 119.7%]), a wide variability was observed in three of eight patients (>30% reduction in bioavailability). Concomitant gastric tube feeding did not affect gatifloxacin bioavailability (interrupted tube feeds: 98.5% [range 61.1% to 119.7%]; continuous tube feeding: 109.0% [range 86.2% to 142.1%]; p =.42). Neither a period nor differential carryover effect was observed. CONCLUSIONS: Although concomitant tube feeding did not affect gatifloxacin bioavailability, critical illness resulted in significant variability that may complicate the role of gatifloxacin in sequential intravenous-to-oral therapy. More research is needed to identify those patients in whom gatifloxacin bioavailability is reduced and for whom an empirical increase in gatifloxacin dose should be considered. PMID- 12771602 TI - Comparison between replacement at 4 days and 7 days of the infection rate for pulmonary artery catheters in an intensive care unit. AB - OBJECTIVE: To compare the relationship between the time of pulmonary artery catheter replacement (4 days or 7 days after insertion) and the occurrence of catheter-associated infections. DESIGN: One-year prospective, randomized, controlled clinical trial. SETTING: Surgical and medical intensive care units at a 2,700-bed medical center. PATIENTS: A total of 258 patients in critical condition who underwent pulmonary artery catheter insertion were recruited. INTERVENTIONS: All patients were randomized into two groups (4 days or 7 days) according to the length of time before the pulmonary artery catheter and pressure monitoring system were replaced. MEASUREMENTS AND MAIN RESULTS: Over a 12-month period, 331 catheters were inserted in 258 patients. In the per-protocol analysis, 98 patients (73.7%) in the 4-day group and 85 patients (68%) in the 7 day group were enrolled. Twelve patients (14.1%) in the 7-day group and 5 patients (5.1%) in the 4-day group (odds ratio, 3.06; 95% confidence interval, 0.94-10.48) had pulmonary artery catheter-tip colonization. Nine patients (10.5%) in the 7-day group and 7 patients (7.1%) in the 4-day group (odds ratio, 1.54; 95% confidence interval, 0.50-4.85) had bacteremia. In the 7-day group, pulmonary artery catheter-related bacteremia was found in only one patient (1.1%, 1.1 episodes per 1,000 catheter-days) compared with no patients in the 4-day group. The frequency of positive cultures from different sources between the 4-day and 7 day groups was not significantly different in the intention-to-treat analysis (p >.05). CONCLUSIONS: No statistically significant difference was found for pulmonary artery catheter-associated infection when intervals of 4 or 7 days between insertion and replacement were compared. Patients with prolonged pulmonary artery catheterization must be carefully examined for signs or symptoms of infection. The time until pulmonary artery catheter replacement can be extended to 7 days if there is no evidence of catheter-related infection. PMID- 12771603 TI - Lipoprotein metabolism in patients with severe sepsis. AB - OBJECTIVE: Lipoproteins have been implicated to play a role in innate immunity. Changes in lipoprotein levels have been reported in a variety of inflammatory disorders. Not much is known about lipoprotein metabolism in patients with severe sepsis. We conducted an ancillary study in a multiple-center phase III sepsis trial to investigate the dynamics of plasma lipoproteins in patients with severe sepsis. DESIGN: Prospective analysis in patients meeting criteria for severe sepsis as part of a multiple-center sepsis study (KyberSept) with antithrombin III (Kybernin P). SETTING: University hospital intensive care unit. PATIENTS: Seventeen patients were included in the study. INTERVENTIONS: Randomized patients received a loading dose of 6000 IU of antithrombin III (Kybernin P) or placebo followed by a 96-hr continuous infusion of 250 IU/hr antithrombin III (Kybernin P) or placebo. In each patient, serial blood samples for total cholesterol, lipoprotein cholesterol, triglycerides, apolipoprotein A-1, apolipoprotein B, and C-reactive protein determination as well as clinical data were collected over 28 days. MEASUREMENTS AND MAIN RESULTS: Plasma cholesterol levels rapidly decreased from 2.67 +/- 2.02 mmol/L on day 0 to a nadir of 1.41 +/- 0.70 mmol/L on day 3, followed by a slow increase to 4.18 +/- 1.94 mmol/L on day 28. High-density lipoprotein (HDL) cholesterol concentrations decreased rapidly from 0.84 +/- 0.92 mmol/L to a nadir of 0.42 +/- 0.35 mmol/L on day 3, to show a slow increase during the following 4 wks to 0.84 +/- 0.42 mmol/L. The low-density lipoprotein (LDL) cholesterol concentrations were already low (0.94 +/- 0.81 mmol/L) at study entry, to show a progressive increase to subnormal values (2.01 +/- 0.94 mmol/L) at 4 wks. Nadir and recovery lipoprotein concentrations were significantly different (paired Student's t-test, p <.05). A significant correlation was found between HDL cholesterol and apolipoprotein A-1 (r =.714, p <.05) and between LDL cholesterol and apolipoprotein B (r =.733, p <.05). There was no statistical difference in lipoprotein concentrations either between survivors and nonsurvivors or between patients receiving antithrombin III or placebo. Serum amyloid A was a major apoprotein (45%) in HDL at the start of the sepsis and was slowly replaced by apolipoprotein A-1 during recovery. A positive correlation was found between plasma C-reactive protein concentrations and serum amyloid A concentrations in HDL (r =.684, p <.05). No other relevant correlations were found between inflammatory and lipoprotein parameters. CONCLUSIONS: In patients with severe sepsis, lipoprotein concentrations rapidly change and can be reduced to 50% of recovery concentrations. The pattern of early rapid decline is found primarily in the HDL and a slow recovery in both HDL and LDL fractions. The correlation between apolipoprotein and lipoprotein cholesterol concentrations suggests a decline in lipoprotein particles. During severe sepsis, HDL is shifted to acute phase HDL, which is enriched in serum amyloid A and depleted of cholesterol and apolipoprotein A-1. Lipoprotein concentrations are unable to discriminate between survivors and nonsurvivors. PMID- 12771604 TI - Heat shock protein 70-2+1267 AA homozygotes have an increased risk of septic shock in adults with community-acquired pneumonia. AB - OBJECTIVE: Heat shock protein (HSP)70-2 is an important immunomodulatory protein induced in response to inflammatory stimuli. We assessed whether HSP70-2+1267 genotype influenced the risk of septic shock in a prospective cohort study of community-acquired pneumonia and whether HSP70-2+1267 genotype is a better predictor of septic shock than the genotype at lymphotoxin-alpha +250. DESIGN: Prospective cohort study. SETTING: A large, nonprofit, private hospital system in Memphis, TN. PATIENTS: Adults admitted with community-acquired pneumonia between 1998 and 2001. Septic shock was defined according to consensus criteria (American College of Chest Physicians/Society of Critical Care Medicine, 1992). INTERVENTIONS: Blood sampling. MEASUREMENTS AND MAIN RESULTS: A total of 343 subjects were enrolled; 30 had septic shock. HSP70-2+1267 and lymphotoxin-alpha +250 genotype was determined using polymerase chain reaction and restriction enzyme digestion. HSP70-2+1267 AA genotype was the strongest predictor of septic shock (p =.0005; relative risk, 3.5). Lymphotoxin-alpha +250 AA genotype was also associated with an increased risk of septic shock (p =.002; relative risk, 2.7). Logistic regression analysis found only age (p =.04) and HSP70-2+1267 genotype (p =.006) were predictors of septic shock. The greatest risk of septic shock was associated with carriage of the HSP70-2+1267 A/lymphotoxin-alpha +250 A haplotype (p <.0001). CONCLUSIONS: HSP70-2+1267 genotype is a stronger predictor of septic shock in patients with community-acquired pneumonia than lymphotoxin-alpha +250 genotype. PMID- 12771605 TI - Morbidity, mortality, and quality-of-life outcomes of patients requiring >or=14 days of mechanical ventilation. AB - OBJECTIVE: To determine the outcome and health-related quality of life of patients requiring >or=14 days of mechanical ventilation in the intensive care unit (ICU). DESIGN: Prospective cohort study with post-ICU, cross-sectional, health-related quality-of-life survey. SETTING: A 17-bed ICU in a university hospital. PATIENTS: A consecutive cohort of 347 patients receiving mechanical ventilation for >or=14 days. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Of the patients enrolled in the study, 150 (44%) died in the ICU and 197 were discharged (58 of 197 died 1-57 months after discharge). Factors associated with ICU death according to multivariate logistic regression analysis were age >or=65 yrs, preadmission New York Heart Association functional class of >or=3, a preadmission immunocompromised status, septic shock at ICU admission, renal replacement therapy in the ICU, and nosocomial septicemia. Cox proportional hazards multivariate analysis identified age of >or=65, a preadmission immunocompromised status, and duration of mechanical ventilation for >35 days as independent predictors of death after ICU discharge. By contrast, postcardiac surgery patients had a better outcome. Health-related quality of life was evaluated for 87 of the 99 long-term survivors after a median follow-up of 3 yrs by using the Nottingham Health Profile and St. George's Respiratory questionnaires. Compared with those of a general French population, their scores were significantly worse for each of the Nottingham Health Profile domains, except social isolation. Nottingham Health Profile scores did not significantly differ between postcardiac and nonpostcardiac surgery patients, men and women (except that women felt more socially isolated), and patients with and without acute respiratory distress syndrome (except for more sleep disorders in those with acute respiratory distress syndrome). Finally, pulmonary-specific St. George's Respiratory Questionnaire global score was worse for acute respiratory distress syndrome survivors. CONCLUSIONS: Prolonged mechanical ventilation is associated with impaired health-related quality of life compared with that of a matched general population. Despite these handicaps, 99% of the patients evaluated were independent and living at home 3 yrs after ICU discharge. Future studies should focus on physical or psychosocial rehabilitation that could lead to improved management of patients after their ICU stay. PMID- 12771606 TI - Adrenocortical hormones in survivors and nonsurvivors of severe sepsis: diverse time course of dehydroepiandrosterone, dehydroepiandrosterone-sulfate, and cortisol. AB - OBJECTIVE: Activation and suppression of immune responses are crucial events during sepsis. Based on substantial new data, a complex picture of differential immune-enhancing and immunosuppressive actions of adrenocortical steroids is emerging. The adrenal androgen dehydroepiandrosterone and its precursor, dehydroepiandrosterone-sulfate, show a considerable decrease with increasing age and serve as functional antagonists to endogenous glucocorticoids. Therefore, we examined time-dependent changes in dehydroepiandrosterone, dehydroepiandrosterone sulfate, cortisol, adrenocorticotropin, and inflammatory variables in surviving and nonsurviving patients with severe sepsis. DESIGN: Prospective observational study in consecutive patients. SETTING: Medical and interdisciplinary intensive care units in two university hospitals and one city hospital. PATIENTS: Thirty nonsurgical patients (25 men and 5 women) with severe sepsis (American College of Chest Physicians/Society of Critical Care Medicine criteria); 15 survivors (mean age, 54 +/- 14 yrs; Acute Physiology and Chronic Health Evaluation III score, 59 +/- 35) and 15 nonsurvivors (mean age, 63 +/- 15 yrs; Acute Physiology and Chronic Health Evaluation III score, 67 +/- 24) were included. Hormones were compared individually and between survivors/nonsurvivors by sequential blood drawings from early sepsis till time of recovery/death. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: During early sepsis, cortisol (nmol/L) was not significantly higher in survivors than nonsurvivors (750 +/- 121 vs. 454 +/- 92, p <.08) and decreased in survivors (p <.01) during late sepsis. During early sepsis, dehydroepiandrosterone-sulfate (percentage of age-matched normal levels) was higher in survivors than nonsurvivors (85 +/- 19 vs. 22 +/- 7, p <.01). Dehydroepiandrosterone-sulfate decreased in survivors (p =.0001) but remained low in nonsurvivors during late sepsis. Dehydroepiandrosterone (percentage of age matched normal levels) was not significantly elevated in survivors compared to nonsurvivors during early sepsis (282 +/- 42 vs. 214 +/- 63, p <.08). Dehydroepiandrosterone decreased in survivors (p <.01) but not in nonsurvivors during late sepsis. Linear regression for dehydroepiandrosterone levels showed a reconstitution of age dependence only in survivors during recovery. Adrenocorticotropin levels did not change. The dehydroepiandrosterone sulfate/cortisol ratio decreased significantly in both survivors and nonsurvivors, whereas dehydroepiandrosterone/cortisol ratio only decreased in survivors during course of sepsis. CONCLUSIONS: During sepsis, adrenal androgens and glucocorticoids show a diverse time-dependent course in survivors and nonsurvivors. PMID- 12771607 TI - Albumin-adjusted calcium is not suitable for diagnosis of hyper- and hypocalcemia in the critically ill. AB - OBJECTIVE: To evaluate whether calcium adjusted for albumin can be used to monitor calcium homeostasis in critically ill patients. DESIGN: Prospective single-single center observational study. SETTING: Clinical laboratory and critical care unit of a regional teaching hospital. PATIENTS: Fifty-three paired samples were from 36 patients requiring intensive care treatment. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Total calcium, albumin-adjusted calcium, and ionized calcium were measured in critically ill patients during an 8-wk period. Calcium was adjusted for albumin using the formula that is most frequently used in The Netherlands. Using ionized calcium as the gold standard, albumin-adjusted calcium overestimated hypercalcemia and totally missed hypocalcemia. The same seemed to be true for other formulas used for albumin or protein adjustment of calcium concentrations. CONCLUSIONS: Albumin-adjusted calcium cannot be used in an intensive care setting to monitor reliably the calcium levels in critically ill patients and should be replaced by measurement of ionized calcium. PMID- 12771608 TI - Ischemic skin lesions as a complication of continuous vasopressin infusion in catecholamine-resistant vasodilatory shock: incidence and risk factors. AB - OBJECTIVE: To report on the incidence and risk factors associated with the development of ischemic skin lesions (ISL) in critically ill patients with catecholamine-resistant vasodilatory shock treated with a continuous infusion of arginine-vasopressin (AVP). DESIGN: Retrospective analysis. SETTING: Twelve-bed general and surgical intensive care unit in a university hospital. PATIENTS: A total of 63 critically ill patients with catecholamine-resistant vasodilatory shock. INTERVENTIONS: Continuous AVP infusion. MEASUREMENTS AND MAIN RESULTS: Demographic, hemodynamic, laboratory data, and skin status were evaluated 24 hrs before and during AVP therapy (24 and 48 hrs). Patients were grouped according to development of new ISL during AVP therapy. A mixed-effects model was used to compare groups. A multiple logistic regression analysis was used to identify independent risk factors for the development of ISL. ISL developed in 19 of 63 patients (30.2%). Thirteen of 19 patients (68%) developed ISL in distal limbs, two patients (10.5%) developed ISL of the trunk, four patients (21%) developed ISL in distal limbs and in the trunk. Five patients (26%) had additional ischemia of the tongue. Body mass index, preexistent peripheral arterial occlusive disease, presence of septic shock, and norepinephrine requirements were significantly higher in patients developing ISL. ISL patients received significantly more units of fresh frozen plasma and thrombocyte concentrates than patients without ISL. Preexistent peripheral arterial occlusive disease and presence of septic shock were independently associated with the development of ISL during AVP therapy. CONCLUSIONS: ISLs are a common complication during continuous AVP infusion in patients with catecholamine-resistant vasodilatory shock. The presence of septic shock and a history of peripheral arterial occlusive disease are independent risk factors for the development of ISL. PMID- 12771609 TI - Usefulness of left ventricular stroke volume variation to assess fluid responsiveness in patients with reduced cardiac function. AB - OBJECTIVE: Stroke volume variation as measured by the analysis of the arterial pressure waveform enables prediction of volume responsiveness in ventilated patients with normal cardiac function. The aim of this study was to investigate the ability of monitoring stroke volume variation to predict volume responsiveness and to assess changes in preload in patients with reduced left ventricular function after cardiac surgery. DESIGN: Prospective study. SETTING: University hospital. PATIENTS: Fifteen mechanically ventilated patients with a left ventricular ejection fraction <0.35 (study group) and 15 patients with an ejection fraction >0.50 (control group) after coronary artery bypass grafting following admission to the intensive care unit. INTERVENTIONS: Volume loading with 10 mL of hetastarch 6% times body mass index. If stroke volume index increased >5%, successive volume loading was performed until no further increase in stroke volume index was reached. MEASUREMENTS AND MAIN RESULTS: Stroke volume variation, central venous pressure, pulmonary artery occlusion pressure (PAOP), and left ventricular end-diastolic area index (LVEDAI) were measured at baseline and immediately after each volume loading step. In both groups, stroke volume variation at baseline correlated significantly with changes in stroke volume index caused by volume loading (p <.01). Further, changes in stroke volume variation as a result of volume loading correlated significantly with the concomitant changes in stroke volume index in both groups (p <.01). Using receiver operating characteristic analysis, in the study group areas under the curve for stroke volume variation, PAOP, central venous pressure, and LVEDAI did not differ significantly. In the control group, the area under the curve for stroke volume variation was statistically larger than for PAOP, central venous pressure, and LVEDAI. CONCLUSIONS: Continuous and real-time monitoring of stroke volume variation by pulse contour analysis can predict volume responsiveness and allows real-time assessment of the hemodynamic effect of volume expansion in patients with reduced left ventricular function after cardiac surgery. PMID- 12771610 TI - Prophylactic anticoagulation with enoxaparin: Is the subcutaneous route appropriate in the critically ill? AB - BACKGROUND: Subcutaneously administered low-molecular-weight heparins are widely used for prevention of venous thromboembolism. The appropriateness of the subcutaneous route in critically ill patients has never been established. OBJECTIVE: To determine anti-Xa activities in critically ill patients and in noncritically ill patients receiving prophylactic doses of subcutaneous enoxaparin. DESIGN: Prospective, controlled, open-labeled study. SETTING: Tertiary medical-cardiologic-postoperative intensive care unit and a general medical ward at a university hospital. PATIENTS: A total of 16 intensive care unit patients (group 1; age, 61.1 +/- 16 yrs; male/female ratio, 7/9; Acute Physiology and Chronic Health Evaluation II score, 20.9 +/- 7; mechanical ventilation, n = 15; vasopressors, n = 13) and 13 noncritically ill medical patients (group 2; age, 61.7 +/- 9 yrs; male/female ratio, 7/6) were studied. Body mass index (25.7 +/- 5 vs. 24 +/- 6 kg/m2, p = not significant) was comparable and serum creatinine levels (0.83 +/- 0.25 vs. 1.07 +/- 0.3 mg/dL, group 1 vs. 2) were within the normal range in both groups. Patients with impaired renal function, receiving hemofiltration, or requiring therapeutic anticoagulation were not eligible. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Anti-Xa activities were determined at 0, 1, 3, 6, and 12 hrs after a single daily subcutaneous dose of 40 mg enoxaparin on day 1 and at 3 hrs after 40 mg of enoxaparin on days 2-5. Mean anti-Xa levels at 0 to 12 hrs were consistently lower in group 1 compared with group 2 by analysis of variance (p =.001 between groups and over time), as was the area under the curve at 0 to 12 hrs (2.6 +/- 1 vs. 4.2 +/- 1.7 units x mL(-1) x hr(-1), group 1 vs. 2, p =.008). Significant differences in anti-Xa activity were also found on days 2-5 (p =.001). Peak anti-Xa activities at 3 hrs after administration were negatively correlated with the body mass index (r = -.41, p <.03). No correlation was found between the anti-Xa activity at 3 hrs and the dose of norepinephrine (r =.12, p =.7). CONCLUSION: Critically ill patients with normal renal function demonstrated significantly lower anti-Xa levels in response to a single daily dose of subcutaneous enoxaparin when compared with medical patients in the normal ward. PMID- 12771611 TI - Intensive care in a field hospital in an urban disaster area: lessons from the August 1999 earthquake in Turkey. AB - OBJECTIVE: To describe our experience with the implementation of intensive care in the setting of a field hospital, deployed to the site of a major urban disaster. DESIGN: Description of our experience during mission to Turkey; conclusions regarding implementation of intensive care at disaster sites. SETTING: Military Field Hospital at Adapazari in Turkey. PATIENTS: Civilian patients admitted for care at the field hospital. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: On August 17, 1999 a major earthquake occurred in western Turkey, causing approximately 16,000 fatalities and leaving >44,000 injured. Approximately 66,000 buildings were severely damaged or destroyed. A medical unit of the Israeli Defense Forces Medical Corps, consisting of 23 physicians, 13 nurses, nine paramedics, 13 medics, laboratory and roentgen technicians, pharmacists, and associated support personnel, were sent to Adapazari in Turkey. The field hospital treated approximately 1,200 patients over a period of 2 wks, 70 surgical operations were performed, 20 babies were delivered, and a variety of medical, surgical, orthopedic, and pediatric/neonatal care was provided. The 12-bed intensive care unit operated by the unit, was staffed by three physicians and eight nursing/paramedic personnel. Patient mix was: a total of 63 patients, among them five with major trauma, 20 with acute cardiac disease, 15 patients with various acute medical conditions, and 11 surgical and postoperative patients. Three patients were intubated and mechanically ventilated (one cardiogenic pulmonary edema and two major trauma). The intensive care unit provided the following functions to the field hospital: care of the critically ill and injured, preparation for and implementation of transportation of such patients, pre- and postoperative care for major surgical procedures, expertise, and equipment for the care of very ill patients throughout the field hospital. CONCLUSIONS: In suitable circumstances, an intensive care capability should be an integral part of medical expeditions to major disasters. PMID- 12771612 TI - Helium/oxygen mixture reduces the work of breathing at the end of the weaning process in patients with severe chronic obstructive pulmonary disease. AB - OBJECTIVE: To test the hypothesis that helium/oxygen mixture can reduce the work of breathing at the end of the weaning process in patients with chronic obstructive pulmonary disease. DESIGN: Prospective, randomized, crossover study. SETTING: Two medical intensive care units at two university tertiary care centers. PATIENTS: Thirteen patients with chronic obstructive pulmonary disease evaluated just before and after extubation. INTERVENTIONS: Helium/oxygen and air/oxygen mixtures were administered sequentially, for 20 mins each, in a randomized order, just before extubation. It was possible to repeat the study after extubation in five patients. MEASUREMENTS AND MAIN RESULTS: Before extubation, the helium/oxygen mixture induced no significant variation in the breathing pattern. By contrast, it reduced the work of breathing from 1.442 +/- 0.718 J/L (mean +/- sd) to 1.133 +/- 0.500 J/L (p <.05). This reduction was explained mainly by a reduction in the resistive component of the work of breathing from 0.662 +/- 0.376 to 0.459 +/- 0.256 J/L (p <.01). We also observed a slight reduction in the intrinsic positive end-expiratory pressure from 2.9 +/- 2.1 cm H(2)O to 2.1 +/- 1.8 cm H(2)O (p <.05). Similar results were also observed after extubation in five patients in whom the repetition of the study was possible. CONCLUSIONS: In spontaneously breathing intubated patients with chronic obstructive pulmonary disease recovering from an acute exacerbation, helium/oxygen mixture reduces the work of breathing as well as intrinsic positive end-expiratory pressure without modifying the breathing pattern. PMID- 12771614 TI - Mechanical ventilation may increase susceptibility to the development of bacteremia. AB - OBJECTIVE: We examined the hypothesis that mechanical ventilation with a potentially injurious strategy would predispose animals to the detrimental effects of subsequent instillation of bacteria. DESIGN: Interventional animal study. SETTING: A university hospital research laboratory. SUBJECTS: Fifty Sprague-Dawley male rats. INTERVENTIONS: Rats were anesthetized and randomized to receive a protective (tidal volume 7 mL/kg, positive end-expiratory pressure 5 cm H(2)O, n = 25) or an injurious ventilatory strategy (tidal volume 21 mL/kg, zero positive end-expiratory pressure, n = 25). Hemodynamics were similar during the 1 hr ventilation period in the two groups. Animals were then disconnected from the ventilator and Pseudomonas aeruginosa was instilled intratracheally before extubation. Thereafter, animals breathed spontaneously; mortality rate was assessed up to 48 hrs, at which time the animals were killed. MEASUREMENTS AND MAIN RESULTS: The 48-hr mortality rate was 28% in the protective group and 40% in the injurious group (p = not significant). A positive bacterial culture from the lung was obtained in 56% of the surviving rats in the low tidal volume group and 67% in the high tidal volume group (p =.059). A positive blood bacterial culture was found in 11% of the low tidal volume group and 33% in the high tidal volume group (p <.05). The absolute bacterial count in the blood was lower in the low tidal volume group compared with the high tidal volume group (p <.05). Concentrations of blood tumor necrosis factor-alpha and macrophage inflammatory protein-2, and lung macrophage inflammatory protein-2 at 48 hrs were significantly higher in the low tidal volume group than in the high tidal volume group. CONCLUSIONS: An injurious ventilatory strategy predisposes animals to subsequent bacteremia associated with an impaired host defense reflected by cytokine response. PMID- 12771613 TI - Beneficial hemodynamic and renal effects of intravenous enalaprilat following coronary artery bypass surgery complicated by left ventricular dysfunction. AB - OBJECTIVE: Angiotensin-converting enzyme inhibitors are an effective therapy for all stages of heart failure due to reduced systolic left ventricular function. Because sufficient data on intravenous angiotensin-converting enzyme inhibitors following coronary artery bypass surgery complicated by postoperative left ventricular dysfunction are unavailable, the efficacy and safety of intravenously administered enalaprilat were evaluated. DESIGN: A placebo-controlled, randomized, double-blind protocol. SETTING: Postoperative intensive care unit at the German Heart Institute Berlin. PATIENTS: Forty patients with a left ventricular ejection fraction <35% following coronary artery bypass surgery on the second postoperative day or after weaning from intra-aortic balloon counterpulsation. INTERVENTIONS: A loading dose of enalaprilat 0.625 mg infused over 1 hr was followed by 5 mg/24 hrs administered continuously for up to 72 hrs. MEASUREMENTS AND MAIN RESULTS: Systemic and pulmonary hemodynamic variables, blood gases, hormonal variables, renal function, and electrolytes were measured before and repeatedly during therapy. Acute effects were as follows: At 1 hr, enalaprilat increased the cardiac index (p <.001), stroke volume index (p <.001), and right ventricular stroke work index (p <.03) compared with placebo, whereas mean arterial pressure (p <.008) and both systemic (p <.001) and pulmonary (p <.02) vascular resistance decreased. Continuous effects were as follows: Over 72 hrs, enalaprilat decreased diastolic pulmonary artery pressure (p <.019), pulmonary artery occlusion pressure (p <.02), and central venous pressure (p <.02). The cardiac and stroke volume indexes were consistently higher in the enalaprilat group, whereas systemic and pulmonary vascular resistances were lower. The arterial blood-pressure lowering effect was blunted and heart rate remained unchanged. Mixed venous oxygenation (p <.02) was higher and arterial oxygenation was not modified. Finally, enalaprilat increased creatinine clearance (p <.002) and decreased creatinine (p <.02) and urea (p <.03). CONCLUSIONS: Intravenous enalaprilat safely and effectively improves cardiac and renal function following coronary artery bypass surgery complicated by postoperative left ventricular dysfunction. PMID- 12771615 TI - Measurement of changes in respiratory mechanics during partial liquid ventilation using jet pulses. AB - OBJECTIVE: To compare the changes in respiratory mechanics within the breathing cycle in healthy lungs between gas ventilation and partial liquid ventilation using a special forced-oscillation technique. DESIGN: Prospective animal trial. SETTINGS: Animal laboratory in a university setting. SUBJECTS: A total of 12 newborn piglets (age, <12 hrs; mean weight, 725 g). INTERVENTIONS: After intubation and instrumentation, lung mechanics of the anesthetized piglets were measured by forced-oscillation technique at the end of inspiration and the end of expiration. The measurements were performed during gas ventilation and 80 mins after instillation of 30 mL/kg perfluorocarbon PF 5080. MEASUREMENTS AND MAIN RESULTS: Brief flow pulses (width, 10 msec; peak flow, 16 L/min) were generated by a jet generator to measure the end-inspiratory and the end-expiratory respiratory input impedance in the frequency range of 4-32 Hz. The mechanical variables resistance, inertance, and compliance were determined by model fitting, using the method of least squares. At least in the lower frequency range, respiratory mechanics could be described adequately by an RIC single-compartment model in all piglets. During gas ventilation, the respiratory variables resistance and inertance did not differ significantly between end-inspiratory and end-expiratory measurements (mean [sd]: 4.2 [0.7] vs. 4.1 [0.6] kPa x L(-1) x sec, 30.0 [3.2] vs. 30.7 [3.1] Pa x L(-1) x sec2, respectively), whereas compliance decreased during inspiration from 14.8 (2.0) to 10.2 (2.4) mL x kPa( 1) x kg(-1) due to a slight lung overdistension. During partial liquid ventilation, the end-inspiratory respiratory mechanics was not different from the end-inspiratory respiratory mechanics measured during gas ventilation. However, in contrast to gas ventilation during partial liquid ventilation, compliance rose from 8.2 (1.0) to 13.0 (3.0) mL x kPa(-1) x kg(-1) during inspiration. During expiration, when perfluorocarbon came into the upper airways, both resistance and inertance increased considerably (mean with 95% confidence interval) by 34.3% (23.1%-45.8%) and 104.1% (96.0%-112.1%), respectively. CONCLUSIONS: The changes in the respiratory mechanics within the breathing cycle are considerably higher during partial liquid ventilation compared with gas ventilation. This dependence of lung mechanics from the pulmonary gas volume hampers the comparability of dynamic measurements during partial liquid ventilation, and the magnitude of these changes cannot be detected by conventional respiratory-mechanical analysis using time-averaged variables. PMID- 12771616 TI - Effect of granulocyte colony-stimulating factor on bleomycin-induced acute lung injury and pulmonary fibrosis. AB - OBJECTIVE: Potentially fatal pulmonary toxicity is a dreaded complication of bleomycin. Increased use of granulocyte colony-stimulating factor in patients receiving chemotherapy has been paralleled by an increased incidence of bleomycin induced pulmonary toxicity. We investigated whether granulocyte colony stimulating factor (25 microg x kg(-1) x day(-1), 4 days) enhanced endotracheal bleomycin-induced (5 mg/kg) acute lung injury and fibrosis in rats. SETTING: University laboratory. SUBJECTS: Sprague-Dawley rats. INTERVENTIONS: We compared the effects of alveolar instillation of bleomycin in rats treated with either granulocyte colony-stimulating factor or saline. MEASUREMENTS AND MAIN RESULTS: Mortality was 25% with bleomycin only and 50% with bleomycin + granulocyte colony stimulating factor. Granulocyte colony-stimulating factor increased alveolar neutrophil recruitment, pulmonary edema, and lung myeloperoxidase activity on day 4. Lung static compliance on day 15 was severely decreased with bleomycin alone and showed a further significant decrease when granulocyte colony-stimulating factor was added (controls, 3.85 +/- 0.14 mL/kPa; bleomycin, 1.44 +/- 0.06 mL/kPa; and bleomycin + granulocyte colony-stimulating factor, 0.65 +/- 0.09 mL/kPa; control vs. bleomycin, p <.0001; and bleomycin vs. bleomycin + granulocyte colony-stimulating factor, p =.0003). Lung morphology with bleomycin + granulocyte colony-stimulating factor showed, in addition to the changes observed with bleomycin alone, four patterns indicating more severe disease: honeycomb foci, pleural thickening with hyaline fibrosis, interstitial granuloma with increased number of macrophages but not neutrophils, and established interstitial fibrosis. Lidocaine, which prevents neutrophil adhesion to endothelial cells, inhibited granulocyte colony-stimulating factor-related exacerbation of acute lung injury (bronchoalveolar lavage fluid cells and pulmonary edema) and pulmonary fibrosis (lung static compliance and morphologic changes). CONCLUSIONS: Granulocyte colony-stimulating factor enhances bleomycin induced lung toxicity by a mechanism that probably involves neutrophils. PMID- 12771617 TI - Tumor necrosis factor as a mediator of cardiac toxicity following snake envenomation. AB - OBJECTIVE: To investigate the possible role of tumor necrosis factor in mediating cardiotoxicity following venom injection in a rat. DESIGN: A randomized controlled experimental study using a Langendorff isolated heart model. SETTING: Animal laboratory. SUBJECTS: Adult male Wistar rats. INTERVENTIONS: The control group (n = 10) was injected with saline only. Each animal in the experimental groups 1-3 (n = 10 each) was injected with Vipera aspis venom 500 microg/kg intramuscularly. Group 1 animals received no additional substance beforehand, group 2 animals were injected intramuscularly with 250 microg of soluble tumor necrosis factor receptor (sTNF-R p55) 15 mins before the venom injection, and group 3 animals were injected intraperitoneally with 40 microg of anti-tumor necrosis factor 60 mins before the venom injection. MEASUREMENTS AND MAIN RESULTS: Cardiac performances were investigated following envenomation. Cardiac histology and myocardial tumor necrosis factor-RNA concentrations were assessed. Serum tumor necrosis factor concentrations rose and peaked 2 hrs following envenomation. A reduction in peak systolic pressures, maximum and minimum change in pressure over time, time-pressure integral, and coronary flow occurred in the venom-only-injected rats compared with controls, whereas blocking tumor necrosis factor activity prevented the deleterious cardiac effects of the envenomation. No histologic changes or increases in myocardial tumor necrosis factor-RNA concentrations were detected. CONCLUSION: These results strongly suggest that systemic release of tumor necrosis factor mediates cardiac toxicity following Vipera aspis envenomation. PMID- 12771618 TI - Intravascular infusion of acid promotes intrapulmonary inducible nitric oxide synthase activity and impairs blood oxygenation in rats. AB - OBJECTIVE: To test the hypothesis that intravascular acid infusion promotes intrapulmonary nitric oxide formation by promoting inducible nitric oxide synthase (iNOS) and inhibiting endothelial nitric oxide synthase (eNOS) expression in rats. DESIGN: Prospective, placebo controlled, randomized laboratory study. SETTING: University laboratory. SUBJECTS: Twelve male Sprague Dawley rats weighing 317 +/- 30 g served as study subjects. All animals were anesthetized, paralyzed, and mechanically ventilated throughout the experiment. INTERVENTIONS: The animals were randomized to receive either 0.1 N hydrochloric acid or 0.9% saline intravenously. The infusions were initially given at a rate of 11 mL/kg/hr for 15 mins and then at a rate of 0.95 mL/kg/hr for the remainder of the experiment. Exhaled nitric oxide concentrations and hemodynamic measurements were monitored throughout the experiment. Lung tissues were harvested for Western blot analysis and immunostaining 4 hrs after starting the intravascular infusion. MEASUREMENT AND MAIN RESULTS: At the end of the experiment, we found more than a four-fold higher concentration of exhaled nitric oxide in the acid-treated animals than in the saline-treated animals (p <.001). Western blot analysis revealed that the acid infusion increased intrapulmonary iNOS concentrations (p <.001), yet it decreased intrapulmonary eNOS concentrations (p =.009). Acid-related lung injury manifested as a decrease in blood oxygen tensions (p =.045) and as an increase in lung homogenate interleukin 6 concentrations (p =.003). CONCLUSIONS: Our results reveal that hydrochloric acid infusion stimulates intrapulmonary nitric oxide formation at least in part by promoting the expression of iNOS. Our findings suggest that correcting acidosis should attenuate iNOS formation. Our data also support the idea that metabolic acidosis itself can lead to impaired intrapulmonary gas exchange and increased expression of pro-inflammatory cytokines such as interleukin-6. Whether the induction of intrapulmonary nitric oxide formation mediates or simply indicates lung injury warrants further investigation. PMID- 12771619 TI - Glottic-modulated lung ventilation during continuous transtracheal gas insufflation: an experimental study. AB - We investigated a new method of pulmonary ventilation that included a minitracheostomy, a reverse thrust catheter to deliver continuous flow of gas to the carina, and a threshold valve to avoid lung overinflation. In six lightly sedated healthy sheep, at a continuous flow of 5, 10, or 15 L/min and a threshold valve of 5, 10, 15, or 20 cm H(2)O, we observed a novel respiratory pattern that was characterized either by active lung inflation followed by passive and prolonged inspiratory hold (mixed pattern) or by an absence of all active inspiratory effort and only passive inflation of the lungs (passive pattern). We correlated airway pressure changes with direct visualization of the glottic opening through a fiberoptic bronchoscope. We measured airway pressures at the level of the carina, the subglottic level, and in the pleural space, and respiratory events were monitored through inductive plethysmography. An increase in continuous flow, threshold valve, or both resulted in 1) an increase in glottic breathing; 2) a decrease in respiratory rate, with a decrease in inspiratory pleural pressure excursion; or 3) an increased inspiratory/expiratory ratio and mean airway pressure. During transtracheal gas insufflation, as in this study, a novel respiratory pattern evolved that was modulated by the glottis, accompanied by a decreased effort of breathing; coughing and swallowing remained, and vocalization remained unimpaired. PMID- 12771620 TI - Methylprednisolone inhibits low-flow hypoxia-induced mitochondrial dysfunction in isolated perfused rat liver. AB - OBJECTIVE: To investigate the mechanism by which methylprednisolone protects the liver from hypoxia-induced injury. DESIGN: Prospective control study using the isolated rat liver. SETTING: Animal research facility. SUBJECTS: Male, fasted, pathogen-free Sprague-Dawley rats. INTERVENTIONS: Low-flow hypoxia was produced by reducing afferent perfusate pressure from 10 to 2.5 cm H(2)O; isolated livers were portally perfused for 2 hrs. MEASUREMENTS AND MAIN RESULTS: We measured mitochondrial membrane potential and hydrogen peroxide production by imaging rhodamine 123 and 2'-7'-dichlorofluorescein fluorescence, respectively. Leakage of mitochondrial enzymes was also monitored by assaying mitochondrial aspartate aminotransferase activity in the outflow perfusate, and the radical-scavenging effect of methylprednisolone was assessed by measuring luminol-dependent hydrogen peroxide chemiluminescence. Apoptosis in liver cells was determined by using terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick-end labeling. Rhodamine 123 fluorescence was significantly diminished in the hypoxic liver, especially in the region of the terminal hepatic venules, which is indicative of membrane depolarization in the mitochondria in those areas. Hypoxia induced mitochondrial dysfunction was indicated by leakage of aspartate aminotransferase into the outflow perfusate, and increased 2'-7' dichlorofluorescein fluorescence indicated increased hydrogen peroxide levels, particularly in the midzone. Pretreatment with 30, 10, or 3 mg/kg of methylprednisolone inhibited the hypoxia-induced mitochondrial membrane depolarization and enzyme leakage, although hydrogen peroxide levels and apoptosis in sinusoidal endothelial cells were unaffected. CONCLUSIONS: Methylprednisolone does not protect the liver from hypoxia-induced injury by suppressing hydrogen peroxide production. Instead, the beneficial effect of methylprednisolone seems to be related to its ability to protect against mitochondrial membrane depolarization under hypoxic conditions. PMID- 12771621 TI - Therapeutic potential for transient inhibition of adenosine deaminase in systemic inflammatory response syndrome. AB - OBJECTIVE: We sought to determine the potential usefulness of 2'-deoxycoformycin (pentostatin), an inhibitor of adenosine deaminase, as a postinsult, or prophylactic treatment for systemic inflammatory response syndrome resulting from fecal peritonitis. DESIGN: Prospective, randomized, controlled experiment. SETTING: Small animal basic science laboratory. SUBJECTS: Male Spague-Dawley rats, weighing 300 to 350 g. INTERVENTIONS: Rats with fecal peritonitis (intraperitoneal cecal slurry) were treated with 1 mg/kg pentostatin intraperitoneally 24 hrs before, or intravenously when signs of illness presented (2 hrs after induction of peritonitis). Signs of illness included tachycardia, tachypnea, and leukopenia. All rats received 50 mL/kg 0.9% saline resuscitative fluid at 2 hrs. MEASUREMENTS AND MAIN RESULTS: Survival to day 6 was 100% in nonseptic sham rats, but 33% in untreated septic rats. In rats given pentostatin either 2 hrs after the insult, or 24 hrs before the insult, 6-day survival improved to 81% and 78%, respectively. Histology revealed diffuse peritonitis, and evidence of systemic inflammatory response syndrome, including local and distant site vascular damage and leukocyte activation. These responses to the septic challenge were abrogated by pentostatin treatment. Return of significant amount of tissue adenosine deaminase activity by 24 hrs and later recovery of white blood cell counts argue against any potential for inappropriate immunosuppression by pentostatin. CONCLUSIONS: These data indicate that the novel use of pentostatin to prevent systemic inflammatory response syndrome secondary to fecal peritonitis shows uncommon promise as a therapeutic tool. All indices of systemic inflammatory response syndrome were abrogated and survival improved when pentostatin was not given until after signs of the illness became manifest. Because protection was afforded with treatment 24 hrs in advance of the inciting insult, pentostatin also has the unique potential for use as a true prophylactic agent. PMID- 12771622 TI - Extracellular glutamate and other amino acids in experimental intracerebral hemorrhage: an in vivo microdialysis study. AB - OBJECTIVE: To determine whether extracellular concentrations of glutamate and other amino acids are significantly elevated after intracerebral hemorrhage and, if so, the temporal characteristics of these changes. Although the role of excitotoxic amino acids, particularly that of glutamate, has been described in ischemic stroke and head trauma, no information exists regarding their possible contribution to the pathogenesis of neuronal injury in intracerebral hemorrhage. DESIGN: Prospective, controlled, laboratory trial. SETTINGS: Animal research laboratory. SUBJECTS: Sixteen anesthetized New Zealand rabbits. INTERVENTION: We introduced intracerebral hemorrhage in each of eight anesthetized New Zealand rabbits by injecting 0.4 mL of autologous blood under arterial pressure into the deep gray matter of the cerebrum. MEASUREMENTS AND MAIN RESULTS: Extracellular fluid samples were collected from the perihematoma region and contralateral (right) hemisphere by in vivo microdialysis at 30-min intervals for 6 hrs. Corresponding samples were similarly collected from both hemispheres in each of eight control animals that underwent needle placement without introduction of a hematoma. Concentrations of amino acids (glutamate, aspartate, asparagine, glycine, taurine, and gamma-aminobutyric acid) in the samples were measured by use of high-pressure liquid chromatography with fluorescence detection. Glutamate concentrations (mean +/- sem) were significantly higher in the hemisphere ipsilateral to the hematoma than in the contralateral hemisphere (92 +/- 22 pg/microL vs. 22 +/- 6 pg/microL) at 30 mins after hematoma creation. A significant increase was observed at 30 mins posthematoma creation in the hemisphere ipsilateral to the hematoma compared with the baseline value. A nonsignificant increase in glutamate concentration persisted in the hemisphere ipsilateral to the hematoma, ranging from 134% to 187% of baseline value between 1 and 5 hrs after hematoma creation. In the hemisphere ipsilateral to the hematoma, a three-fold increase in the concentration of glycine was observed at 30 mins after hematoma creation compared with the baseline level (890 +/- 251 pg/microL vs. 291 +/- 73 pg/microL). There was a significant difference between the hemisphere ipsilateral to the hematoma compared with the ipsilateral (corresponding) hemisphere of the control group at 30 mins posthematoma (890 +/- 251 pg/microL vs. 248 +/- 66 pg/microL). A similar transient increase was observed in taurine and asparagine concentrations at 30 mins after hematoma creation, compared with baseline measurements. Taurine concentrations in the hemisphere ipsilateral to the hematoma were significantly higher than the ipsilateral hemisphere of the control group (622 +/- 180 pg/microL vs. 202 +/- 64 pg/microL) at 30 mins after hematoma creation. CONCLUSIONS: The present study suggests that glutamate and other amino acids accumulate transiently in extracellular fluids in the perihematoma region during the early period of intracerebral hemorrhage. The exact role of these amino acids in the pathogenesis of neuronal injury observed in intracerebral hemorrhage needs to be defined. PMID- 12771623 TI - Pathogenic role of interleukin-6 in the development of sepsis. Part I: Study in a standardized contact burn murine model. AB - OBJECTIVE: To establish a representative model for the evaluation of interleukin (IL)-6 and IL-6 receptor for pathogenicity and lethality in the postburn period. DESIGN: Ten-week-old C 57 BL/6J mice received a 20% body surface area contact burn and/or lipopolysaccharide (LPS) 48 hrs later. Standardized burns were created with a metal stamp of 150 degrees C of defined pressure and surface area (2.4525 Newton/0.00166 m2) over a period of 11 secs. The depth of dermal injury was verified histologically. The following groups were formed: I: no burn, no LPS (n = 35); II: burn, no LPS (n = 140); III, no burn, LPS (n = 56); and IV, burn, LPS (n = 80), to study the effect of burn alone, sepsis alone, or the combination. Lethal LPS dose (LD100) was determined by application of LPS in increasing doses (200, 300, 400, and 500 microg, n = 32) after burns. MEASUREMENTS: Concentrations of interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma), and leukocytes, platelets and organ pathology were evaluated. SETTING: Research laboratory. RESULTS: Burn and LPS showed an additive effect on the release of IL-6 but not of TNF-alpha and IFN gamma. Leukocyte and platelet numbers decreased significantly (group IV) compared with the other groups (I-III). The maximal levels of IL-6 in group IV were reached earlier than those of TNF-alpha. The contact burn model has a mortality rate of 30%, which is close to clinical outcome. We found the model of contact burn superior to scald or flame burn models. A dose of 400-microg LPS was found to be the lethal LPS dose (LD100). CONCLUSIONS: Our data suggest that preexisting burn injury increases the response to endotoxin. TNF-alpha is not involved in priming. IL-6 on the other hand is a very representative parameter for priming. Because TNF-alpha was obviously not the causative factor, it was concluded that the application of anti-IL-6-mAb should be of great value. Therefore, a therapeutic application was designed, see part II. PMID- 12771624 TI - Pathogenic role of interleukin-6 in the development of sepsis. Part II: Significance of anti-interleukin-6 and anti-soluble interleukin-6 receptor-alpha antibodies in a standardized murine contact burn model. AB - OBJECTIVE: The in vivo effects of anti-interleukin-6 (anti-IL-6) and anti interleukin-6-alpha receptor (anti-IL-6R) monoclonal antibodies on immune response and survival rate of a burn with subsequent infection were assessed. SUBJECTS: Ten-week-old C 57 BL/6J mice received a standardized contact burn; 48 hrs later endotoxin (LPS) was injected intraperitoneally to induce systemic inflammation. Ten different groups were studied. Groups I-IV sustained a burn and/or a LPS-stimulus but did not receive any anti-cytokines and served as controls. Treatment groups V-X sustained the same injuries but also received anti IL-6 and anti-IL-6R intravenously either before or after the LPS stimulus. In a further part of the study, a lethal dose of LPS was injected (LPS-LD(100) group) followed by an injection of anti-IL-6 antibody and/or anti-IL-6R antibody. MEASUREMENTS: Serum concentrations of IL-6, tumor necrosis factor (TNF)-alpha, interferon (IFN)-gamma, and white blood cell and platelet counts were determined, and the survival rate over a 2-wk period was assessed. RESULTS: Treatment with anti-IL-6 slightly decreased the inflammatory response when it was given before or after LPS application. The inflammatory response was not decreased after treatment with anti-IL-6R. In the groups that received a combination of anti-IL-6 and anti-IL-6R, there was a significant reduction of the inflammatory response. This was more pronounced when the anti-cytokines were applied after LPS application. A significant reduction in mortality could be shown with both antibodies in the treatment groups and the groups that had received a lethal dose of LPS (LPS-LD(100) group). CONCLUSIONS: IL-6 has a low inflammatory potential, and IL-6R has no inflammatory potential by itself. In contrast, the IL-6/IL-6R complexes have a higher inflammatory potential. Mortality could be reduced by each antibody alone as well as by the combination, supporting the hypothesis that the inflammatory and lethal potentials of IL-6 are not identical. The study suggests that the use of antibodies against IL-6 or IL-6R is effective in the prevention of systemic inflammation in a murine burn model. PMID- 12771625 TI - Effects of titrated arginine vasopressin on hemodynamic variables and oxygen transport in healthy and endotoxemic sheep. AB - OBJECTIVE: To determine the effects of titrated arginine vasopressin (AVP) alone or in combination with norepinephrine (NE) on hemodynamics and oxygen transport in healthy and endotoxemic sheep. DESIGN: Prospective controlled trial. SETTING: University research laboratory. SUBJECTS: Six adult ewes. INTERVENTIONS: Healthy sheep received AVP as a titrated infusion, initiated with 0.6 units/hr and increased by 0.6 units/hr every 15 mins, either until mean arterial pressure was increased by 20 mm Hg vs. baseline or a maximum of 3.6 units/hr was administered. After 90 mins, AVP infusion was continued with the investigated dosage, and NE (0.2 microg x kg(-1) x min(-1)) was also infused for 90 mins. After a 24-hr period of recovery, endotoxemia was induced and maintained (Salmonella typhosa endotoxin, 10 ng x kg(-1) x min(-1)) in the same sheep for the next 19 hrs. After 16 hrs of endotoxemia, AVP and NE were administered as described previously. MEASUREMENTS AND MAIN RESULTS: Hemodynamics were obtained at baseline, every 15 mins during the titration period, and 60 and 90 mins after additional NE infusion. Variables of oxygen transport were calculated before and after the titration period. In healthy and endotoxemic sheep, AVP reduced heart rate and cardiac index (p <.001) and compromised oxygen delivery (p <.001) and oxygen consumption (healthy sheep, p =.003; endotoxemic sheep, p <.001). Vasopressin infusion did not alter mean pulmonary arterial pressure but increased pulmonary vascular resistance index in both groups (p <.001). Additional infusion of NE further augmented mean arterial pressure and increased cardiac index during endotoxemia (p <.001). This was accompanied by an increase in oxygen delivery and consumption (p <.05 each). CONCLUSIONS: During ovine endotoxemia, AVP decreased cardiac index, compromised oxygen delivery, and increased pulmonary vascular resistance index. These side effects may limit its use as a sole vasopressor during sepsis. Potentially, a simultaneous infusion of AVP and NE could represent a useful therapeutic option. PMID- 12771626 TI - Species-specific modulation of the nitric oxide pathway after acute experimentally induced endotoxemia. AB - OBJECTIVE: The derangement of the nitric oxide pathway is an important contributing factor to the pathogenesis of septic shock. The aim of this study was to investigate potential differences in modulation of such a pathway in two experimental models of endotoxemia. DESIGN: Prospective, randomized, placebo controlled animal investigation. SETTING: Cardiovascular research laboratory. SUBJECTS: Male, anesthetized, and mechanically ventilated New-Zealand rabbits (n = 24) and Sprague-Dawley rats (n = 24). INTERVENTIONS: After pretreatment with 1400W (1 mg kg(-1) subcutaneously), an inhibitor of inducible nitric oxide synthase, animals received an intravenous bolus of Escherichia Coli lipopolysaccharides (5 mg kg(-1)). After 4 hrs, lungs, myocardial left ventricles, and aortas were collected. MEASUREMENTS AND MAIN RESULTS: Blood mean arterial pressure, pH, and nitrite/nitrate were monitored. Nitric oxide in the exhaled air was measured by chemiluminescence. Tissue activity of both constitutive nitric oxide synthase and inducible nitric oxide synthase was determined by measuring the conversion of [3H]L-arginine to [3H]L-citrulline. In lipopolysaccharide-treated animals, both mean arterial pressure (after 60 to 90 mins) and blood pH (after 4 hrs) decreased with respect to baseline values. 1400W prevented lipopolysaccharide-induced hypotension only in rats (p <.01). Exhaled nitric oxide decreased in lipopolysaccharide-treated rabbits by 120 mins (from 12.6 +/- 0.6 to 8.4 +/- 0.6 ppb, p <.01) and remained low until the end of the experiment (p <.01 vs. baseline). Conversely, exhaled nitric oxide increased in lipopolysaccharide-treated rats by 120 mins (from 0.4 +/- 0.1 to 5.3 +/- 1.7 ppb, p <.01) and reached a plateau by 210 mins (19.8 +/- 3.1 ppb, p <.01 vs. baseline). 1400W prevented the lipopolysaccharide-induced increase in exhaled nitric oxide and blood nitrite/nitrate in rats (p <.05). Inducible nitric oxide synthase activity increased in endotoxemic rabbit heart (0.19 +/- 0.05 vs. 0.07 +/- 0.02 pmol L-citrulline/min/mg protein in the control group, p <.05) and in all rat tissues, being more striking in the lungs (25.00 +/- 0.01 vs. 0.19 +/- 0.04 pmol L-citrulline/min/mg protein in the control group, p <.001). CONCLUSIONS: The nitric oxide pathway is differently modulated between endotoxemic rabbits and rats. PMID- 12771627 TI - Effects of albumin and Ringer's lactate on production of lung cytokines and hydrogen peroxide after resuscitated hemorrhage and endotoxemia in rats. AB - RATIONALE AND HYPOTHESIS: Acute lung injury is a frequent complication of severe sepsis or blood loss and is often associated with an excessive inflammatory response requiring mechanical ventilation. We tested the hypothesis that the types of fluids used during early resuscitation have an important effect on the evolution of lung injury. METHODS: Rats were subjected to either hemorrhage or endotoxemia for 1 hr, followed by resuscitation to a controlled mean blood pressure with Ringer's lactate, 5% albumin, or 25% albumin for 1 hr. After resuscitation, blood cytokine levels were measured. The lung was then excised and ventilated with a tidal volume of 30 mL/kg for 2 hrs. RESULTS: The volume of fluids required was significantly smaller in the albumin-treated groups than in the Ringer's lactate groups. In the hemorrhagic shock model, plasma concentrations of tumor necrosis factor-alpha, interleukin-6, and macrophage inflammatory protein-2 were significantly lower and interleukin-10 was significantly higher in the albumin-treated groups compared with the Ringer's lactate-treated group. The levels of tumor necrosis factor-alpha and macrophage inflammatory protein-2 in bronchoalveolar lavage fluid were lower and interleukin 10 was higher in the albumin-treated groups than in the Ringer's lactate group. The decreased cytokine production was associated with a reduction of hydrogen peroxide formation with albumin resuscitation. The lung wet/dry ratio was lower in the 5% albumin (0.54 +/- 0.01) and 25% albumin (0.55 +/- 0.02) groups than in the Ringer's lactate group (0.62 +/- 0.02; both p <.05). These effects of albumin seen in the hemorrhagic shock model were not observed in the endotoxic shock model. CONCLUSIONS: We conclude that resuscitation with albumin may have utility in reducing ventilator-induced lung injury after hemorrhagic shock, but not after endotoxic shock. These findings suggest that the mechanisms leading to ventilator induced lung injury after hemorrhage differ from those after endotoxemia. PMID- 12771628 TI - Survival without brain damage after clinical death of 60-120 mins in dogs using suspended animation by profound hypothermia. AB - OBJECTIVES: This study explored the limits of good outcome of brain and organism achievable after cardiac arrest (no blood flow) of 60-120 mins, with preservation (suspended animation) induced immediately after the start of exsanguination cardiac arrest. DESIGN: Prospective experimental comparison of three arrest times, without randomization. SETTING: University research laboratory. SUBJECTS: Twenty-seven custom-bred hunting dogs (17-25 kg). INTERVENTIONS: Dogs were exsanguinated over 5 mins to cardiac arrest no-flow of 60 mins, 90 mins, or 120 mins. At 2 mins of cardiac arrest, the dogs received, via a balloon-tipped catheter, an aortic flush of isotonic saline at 2 degrees C (at a rate of 1 L/min), until tympanic temperature reached 20 degrees C (for 60 mins of cardiac arrest), 15 degrees C (for 60 mins of cardiac arrest), or 10 degrees C (for 60, 90, or 120 mins of cardiac arrest). Resuscitation was by closed-chest cardiopulmonary bypass, postcardiac arrest mild hypothermia (tympanic temperature 34 degrees C) to 12 hrs, controlled ventilation to 20 hrs, and intensive care to 72 hrs. MEASUREMENTS AND MAIN RESULTS: We assessed overall performance categories (OPC 1, normal; 2, moderate disability; 3, severe disability; 4, coma; 5, death), neurologic deficit scores (NDS 0-10%, normal; 100%, brain death), regional and total brain histologic damage scores at 72 hrs (total HDS >0-40, mild; 40-100, moderate; >100, severe damage), and morphologic damage of extracerebral organs. For 60 mins of cardiac arrest (n = 14), tympanic temperature 20 degrees C (n = 6) was achieved after flush of 3 mins and resulted in two dogs with OPC 1 and four dogs with OPC 2: median NDS, 13% (range 0-27%); and median total HDS, 28 (range, 4-36). Tympanic temperature of 15 degrees C (n = 5) was achieved after flush of 7 mins and resulted in all five dogs with OPC 1, NDS 0% (0-3%), and HDS 8 (0-48). Tympanic temperature 10 degrees C (n = 3) was achieved after flush of 11 mins and resulted in all three dogs with OPC 1, NDS 0%, and HDS 16 (2-18). For 90 mins of cardiac arrest (n = 6), tympanic temperature 10 degrees C was achieved after flush of 15 mins and resulted in all six dogs with OPC 1, NDS 0%, and HDS 8 (0 37). For 120 mins of cardiac arrest (n = 7), three dogs had to be excluded. In the four dogs within protocol, tympanic temperature 10 degrees C was achieved after flush of 15 mins. This resulted in one dog with OPC 1, NDS 0%, and total HDS 14; one with OPC 1, NDS 6%, and total HDS 20; one with OPC 2, NDS 13%, and total HDS 10; and one with OPC 3, NDS 39%, and total HDS 22. CONCLUSIONS: In a systematic series of studies in dogs, the rapid induction of profound cerebral hypothermia (tympanic temperature 10 degrees C) by aortic flush of cold saline immediately after the start of exsanguination cardiac arrest-which rarely can be resuscitated effectively with current methods-can achieve survival without functional or histologic brain damage, after cardiac arrest no-flow of 60 or 90 mins and possibly 120 mins. The use of additional preservation strategies should be pursued in the 120-min arrest model. PMID- 12771629 TI - Surfactant kinetics in preterm infants on mechanical ventilation who did and did not develop bronchopulmonary dysplasia. AB - OBJECTIVE: To characterize surfactant kinetics in vivo in two groups of premature infants on different levels of mechanical ventilation and at different risk of developing bronchopulmonary dysplasia. DESIGN: Controlled observational study in two independent groups of infants. SETTING: Neonatal intensive care unit. PATIENTS: Thirteen preterm infants (26 +/- 0.5 wks, birth weight 801 +/- 64 g) on high ventilatory setting and who finally all developed bronchopulmonary dysplasia (MechVentBPD), and eight (26 +/- 0.5 wks, birth weight 887 +/- 103 g) who had minimal or no lung disease and of whom none developed bronchopulmonary dysplasia (MechVentNoBPD). MEASUREMENTS AND MAIN RESULTS: Endotracheal 13C-labeled dipalmitoyl-phosphatidylcholine was administered and subsequent measurements of the 13C enrichment of surfactant-disaturated phosphatidylcholine (DSPC) from serial tracheal aspirates were made by gas chromatography-mass spectrometry. We calculated disaturated phosphatidylcholine pharmacokinetic variables in terms of half-life and apparent pool size from the enrichment decay curves over time. DSPC concentration from tracheal aspirates was expressed as milligrams/milliliter epithelial lining fluid (ELF-DSPC). Data are presented as mean +/- se. In MechVentBPD infants vs. MechVentNoBPD, ELF-DSPC was much reduced, 2.9 +/- 0.6 vs. 9.4 +/- 3.0 mg/mL ELF (p =.03), half-life was shorter, 19.4 +/- 2.8 vs. 42.5 +/- 6.3 hrs (p =.002), and apparent pool size larger, 136 +/- 21 vs. 65.8 +/- 16.0 mg/kg (p =.057). In MechVentBPD, apparent DSPC pool size positively correlated with mean airway pressure x Fio(2) and inversely correlated with ELF-DSPC. ELF DSPC was inversely correlated with mean airway pressure x Fio(2). No significant correlations were found in the MechVentNoBPD group. CONCLUSIONS: MechVentBPD infants showed profound alteration of surfactant kinetics compared with preterm infants with minimal lung disease, and these alterations were correlated with severity of ventilatory support. PMID- 12771630 TI - Intracranial pressure monitoring and case mix-adjusted mortality in intracranial hemorrhage. AB - OBJECTIVE: Intracranial pressure (ICP) monitoring is frequently used in intensive care treatment of patients with intracranial hemorrhage. Data demonstrating an improved outcome from this intervention are lacking. We analyzed standardized mortality ratios in patients with and without ICP monitoring to determine its efficacy. DESIGN: A nonrandomized study of case records of consecutively admitted intensive care unit (ICU) patients with intracranial hemorrhage. SETTING: General and medical ICU of a 900-bed tertiary-care hospital. PATIENTS: A total of 225 patients with intracranial hemorrhage (mainly nontraumatic) admitted consecutively between April 1997 and March 2000. MEASUREMENTS: Simplified Acute Physiology Score (SAPS) II, diagnosis, age, sex, use of ICP monitoring, and in hospital mortality rates were collected from the hospital's ICU database. Expected mortality was provided by means of SAPS II. Standardized mortality ratios were calculated and compared in 119 patients with ICP monitoring and 106 patients without ICP monitoring. MAIN RESULTS: The case mix-adjusted hospital mortality in the group with ICP monitoring was in the expected range (standardized mortality ratio, 1.09 [95% confidence interval (CI), 0.87-1.31]). Patients without ICP monitoring had a significantly higher standardized mortality ratio than expected (1.26 [95% CI, 1.06-1.46]). CONCLUSIONS: A beneficial effect of ICP monitoring in patients with intracranial hemorrhage may be reflected in an improved standardized mortality ratio. PMID- 12771631 TI - Do-not-resuscitate order after 25 years. AB - BACKGROUND: In 1976, the first hospital policies on orders not to resuscitate were published in the medical literature. Since that time, the concept has continued to evolve and evoke much debate. Indeed, few initials in medicine today evoke as much symbolism or controversy as the Do-Not-Resuscitate (DNR) order. OBJECTIVE: To review the development, implementation, and present standing of the DNR order. DESIGN: Review article. MAIN RESULTS: The DNR order concept brought an open decision-making framework to the resuscitation decision and did much to put appropriate restraint on the universal application of cardiopulmonary resuscitation for the dying patient. Yet, even today, many of the early concerns remain. CONCLUSIONS: After 25 yrs of DNR orders, it remains reasonable to presume consent and attempt resuscitation for people who suffer an unexpected cardiopulmonary arrest or for whom resuscitation may have physiologic effect and for whom no information is available at the time as to their wishes (or those of their surrogate). However, it is not reasonable to continue to rely on such a presumption without promptly and actively seeking to clarify the patient's (or surrogate's) wishes. The DNR order, then, remains an inducement to seek the informed patient's directive. PMID- 12771632 TI - Surgeons, intensivists, and the covenant of care: administrative models and values affecting care at the end of life--Updated. AB - CONTEXT: End-of-life care remains a challenging and complex activity in critical care units. There is little information concerning the influence of administrative models of care delivery on end-of-life care. OBJECTIVE: To compare and contrast end-of-life care delivery in intensive care units using "semiclosed," "open," and "closed" administrative models. DESIGN: Ethnographic study of three critical care units. SETTING: University hospitals in the United States and New Zealand. SUBJECTS: Approximately 600 physicians, nurses, allied health personnel, patients, family members, and friends. MEASUREMENTS AND MAIN RESULTS: Ethnographic observations were made at three sites for 75, 3, and 10 wks, respectively. Eighty end-of-life care episodes were observed. The interactions among care personnel and families varied according to the administrative model, depending on whether surgeons or intensivists had primary patient responsibility. This led to differential timing on the shift from "cure" to "comfort," and differential decision-making power for families. CONCLUSIONS: End-of-life care varies according to the administrative model. When surgeons have primary responsibility for the patient, the most important goal is defeating death. When intensivists have sole patient responsibility, scarce resources are considered and quality of life is a significant variable. Discussions about improving the way end-of-life decisions are carried out in intensive care units rarely consider the administrative models and personal, professional, and national values affecting such decisions. To improve care at the end of life, we must critically examine these features. PMID- 12771633 TI - Measures, markers, and mediators: toward a staging system for clinical sepsis. A report of the Fifth Toronto Sepsis Roundtable, Toronto, Ontario, Canada, October 25-26, 2000. AB - BACKGROUND: Sepsis is not a single disease but a complex and heterogeneous process. Its expression is variable, and its severity is influenced by the nature of the infection, the genetic background of the patient, the time to clinical intervention, the supportive care provided by the clinician, and a number of factors as yet unknown. The evaluation of effective therapies has been hampered by limitations in our ability to characterize the process and to stratify patients into more homogeneous groups with respect to pathogenesis. OBJECTIVES: To develop a taxonomy of markers relevant to clinical research in sepsis and to propose a testable candidate system for stratifying patients into more therapeutically homogeneous groups. DATA SOURCE: An expert roundtable discussion and a MEDLINE review using search terms "marker" and "sepsis." RESULTS: Markers provide information in one or more of three domains: diagnosis, prognosis, and response to therapy. More than 80 putative markers of sepsis have been described. All correlate with the risk of mortality (prognosis), yet none has shown utility in stratifying patients with respect to therapy (diagnosis) or in titrating that therapy (response). Their limitations arise from the challenges of establishing causality in a complex disease process such as sepsis and of stratifying patients into more homogeneous populations. The former limitation may be addressed through a modification of Koch's postulates to differentiate causality from simple association. The latter suggests the need for a staging system analogous to those used in other complex disease processes such as cancer. A candidate framework for such a system, based on the infection, the host response, and the extent of organ dysfunction (the IRO system) is described. CONCLUSIONS: Advances in the understanding and management of patients with sepsis will necessitate more rigorous approaches to disease description and stratification. Models should be developed, tested, and modified through clinical studies rather than through consensus. PMID- 12771634 TI - The physician's role in discussing organ donation with families. AB - Federal Conditions of Participation from the Health Care Financing Administration (now the Centers for Medicare and Medicaid Services) introduced in 1998 require that all families be presented the option of organ and tissue donation when death is imminent. The perception that physicians were being excluded from participating in this process led to a resolution at the American Medical Association House of Delegates meeting in December 1999, calling on the American Medical Association Council on Scientific Affairs to review the Conditions of Participation "to ensure that there is no prohibition of physician involvement in the organ donation process..." The number of organs procured for transplantation in the United States is insufficient to meet needs. Families' hospital experiences significantly affect their decisions to donate organs. Discussing severe brain injury, brain death, and organ donation after brain death with families is a specialized form of end-of-life decision-making and care in the intensive care unit; however, the knowledge, skills, and attitudes necessary for physicians and nurses to promote good end-of-life decision-making are widely variable. The federal Conditions of Participation require that those making requests of families for organ donation receive specific training. They do not prohibit physician involvement in initiating organ donation requests, provided these individuals are properly trained. Physicians have an important role in caring for patients and families in these circumstances, and the care they provide is enhanced through training, attention to the special issues involved, and collaboration with organ procurement organization personnel. PMID- 12771635 TI - Tracheal stenosis and obliteration above the tracheostoma after percutaneous dilational tracheostomy. AB - Percutaneous dilational tracheotomy (PDT) as opposed to the conventional surgical tracheostomy is a procedure that allows airway control in critically ill patients without surgical exposure of the trachea. Based on the Seldinger technique, dilators are passed along a guiding wire through a small neck incision into the trachea under endoscopic surveillance. This separates the tracheal rings and results in a stoma. As opposed to the regular surgical tracheostoma, a PDT-stoma is not epithelialized. The procedure is cost effective and little time consuming. Considering the increasing number of performed PDTs in the last few years, we feel a need to be aware of possible long-term complications. Thus, in this report, we describe three cases of tracheal stenosis/obliteration after a PDT procedure. In all cases, tracheal narrowing occurred above the level of the stoma. This suggests a procedure-related mechanism, i.e., tracheal ring invagination and the consecutive development of granulation tissue, rather than a mechanism based on the duration of the cannula's placement, which would normally produce the stenosis below the stoma in the area of the cuff. Toward the end of the article, we provide evidence for this hypothesis and thus present a new subset of long-term complications after PDT. PMID- 12771636 TI - Successful treatment of Candida krusei infection with caspofungin acetate: a new antifungal agent. AB - OBJECTIVE: Systemic fungal infections have high mortality, and therapy is often toxic. Caspofungin acetate, a new antifungal agent with minimal toxicity, may provide a better alternative to typical therapy for Candida krusei. DESIGN: Case report. SETTING: Multidisciplinary intensive care unit (ICU) of a community teaching hospital. PATIENT: A 22-yr-old male with acute lymphoblastic leukemia and Candida krusei fungemia failed therapy with fluconazole and amphotericin B. INTERVENTIONS: Caspofungin acetate given intravenously as a 70-mg loading dose, followed up by 50 mg daily along with standard ICU care. RESULTS: Survival without toxicity from therapy. CONCLUSION: Efficacy of caspofungin acetate in a patient with life-threatening Candida Krusei infection. PMID- 12771637 TI - Bone marrow transplantation and intensive care unit admission: what really matters? PMID- 12771638 TI - Difficult to wean chronic obstructive pulmonary disease patients: avoid heat and moisture exchangers? PMID- 12771639 TI - Dollars and sense in the intensive care unit: the costs of ventilator-associated pneumonia. PMID- 12771640 TI - Habit and the hub. PMID- 12771641 TI - The long and short of pulmonary artery catheter monitoring and catheter-related infections: is less still best? PMID- 12771642 TI - More than skin deep? Complications associated with vasopressin infusion: a cautionary tale. PMID- 12771643 TI - A bioavailability study in the proposed patient population--with much more needed now. PMID- 12771644 TI - Intensive care in a field hospital in an urban disaster area: are we ready? PMID- 12771645 TI - Is there an "ACE" in the hole for postcoronary artery bypass graft myocardial dysfunction? PMID- 12771646 TI - Desperate appliance. PMID- 12771647 TI - Do not resuscitate: ordering nonassault and charting patients' decisions to forgo cardiopulmonary resuscitation. PMID- 12771648 TI - Physicians and organ procurement organizations--an essential partnership that must not be neglected. PMID- 12771649 TI - Family satisfaction with intensive care unit care: influenced by workload, staffing, and patient selection? PMID- 12771650 TI - Activated protein C: more effective than nonactivated protein C? PMID- 12771651 TI - Avoiding biased publishing. PMID- 12771652 TI - Sedation in pediatric patients. PMID- 12771653 TI - Selective decontamination of the digestive tract: use in liver transplantation is evidence based. PMID- 12771654 TI - Administration of methylene blue in human septic shock: renaissance of an old drug? PMID- 12771655 TI - General anesthesia or sedation in the intensive care unit: are we aware of the difference when we use propofol? PMID- 12771656 TI - Which came first? PMID- 12771657 TI - A technique that improves the safety of feeding tube insertion. PMID- 12771658 TI - The last bifocal trial? PMID- 12771659 TI - Ocular manifestations of Donohue's syndrome. AB - INTRODUCTION: Donohue's syndrome, also known as Leprechaunism, is a rare autosomal recessive disease that manifests at birth with symptoms of endocrine dysfunction. Metabolic characteristics of the disease include postprandial hyperglycemia, fasting hypoglycemia, insulin resistance, hyperinsulinemia, and failure to thrive. The physical features most often associated with this condition include hypertrichosis, pachyderma, acanthosis nigricans, prominent genitalia, and elfin-like facial characteristics of prominent eyes, wide nostrils, thick lips, and large, low-set ears. Not only is this syndrome rare, but it often results in infant and early childhood mortality. The literature regarding ocular manifestations is limited. CASE REPORT: We present a case of a 29-year-old male with Donohue's syndrome and significant ocular findings including a subluxated mature cataract, retinal detachment, high myopia, and optic atrophy. DISCUSSION: These ocular sequelae are discussed with regard to the noted endocrine dysfunction and its effects on tissue development and growth. PMID- 12771660 TI - Uveal melanoma and poor treatment compliance: an atypical outcome with literature review. AB - BACKGROUND: A uveal melanoma is the most common primary intraocular malignancy and is the primary intraocular disease that can be fatal in adults. Until recently, data were limited regarding growth characteristics and at what stage metastases are likely to occur. CASE REPORT: A 49-year-old white male presented on January 19, 1989, for a second opinion regarding a spot that had been discovered in the right eye 13 months prior. Ophthalmologic evaluation and echography were consistent for a uveal melanoma of 5 mm elevation and 13.5 x 6 mm in diameter. The patient refused treatment or enrollment into the Collaborative Ocular Melanoma Study. On December 9, 1991, the eye was enucleated with a modified exenteration due to severe proptosis, tumor thickness of 22 mm, and three areas of extrascleral extension. The patient refused postoperative radiation treatment and refused clinical follow-up. This case was reviewed from the initial evaluation date through the last clinical visit. The patient was recently interviewed with respect to his use of homeopathic medicine, change in lifestyle, and the effect he felt it might have had on his life. DISCUSSION: The literature published by the Collaborative Ocular Melanoma Study regarding the histopathologic characteristics of eyes enucleated and the initial mortality findings of a large choroidal melanoma indicate that this is a most unusual case. The patient has defied the mortality findings of the most recent and largest study to date (mean survival rate, 82 months). The patient is still alive 181 months after the original finding of the lesion and refusal of advised treatment recommendations in favor of homeopathic medicine. PMID- 12771661 TI - Transcranial magnetic stimulation as an investigative tool in the study of visual function. AB - Transcranial magnetic stimulation (TMS) is a novel and powerful probe to study the relationship between human brain function and behavior. TMS is being widely used to investigate memory, language, attention, learning, and motor function and is even being utilized therapeutically in the treatment of depression. Some of the earliest applications of TMS have been directed toward the investigation of human visual perception. For example, a strong TMS pulse delivered to the occipital cortex in a sighted or even blind individual can evoke the sensation of perceiving light (visual phosphenes). TMS can also be used to suppress visual perception and investigate the timing of visual information processing. Furthermore, the functional connectivity between different brain areas can be mapped using TMS, thus establishing a causal link between visual cortical function and visual perception. The present article is meant as an overview of the technique of TMS and a review of the literature as it pertains to the study of visual function. The application of TMS in the diagnosis as well as possible therapeutic use in various visual disorders is also discussed. PMID- 12771662 TI - Prevalence of myopia among 12- to 13-year-old schoolchildren in northern Mexico. AB - PURPOSE: The aim of this article was to report the prevalence of refractive errors, mainly myopia, among 12- to 13-year-old children in a metropolitan setting in Mexico. METHODS: A total of 1035 schoolchildren were examined in a field study in Monterrey, Mexico. The examination included best-corrected visual acuity and refraction during cycloplegia. A sample of the children was sent to a pediatric eye clinic and underwent cycloplegic refraction with an autorefractor. RESULTS: We found a prevalence of myopia (>/=-0.5 D SE) of 44%, whereas bilateral myopia was present in 37% of the children. In the total sample, high myopia (>/= 5D) was found in 1.4%. The prevalence of myopia was significantly higher in girls. Only 20% of children with bilateral myopia used prescription glasses; 8% had prescribed glasses, but did not use them. Hyperopia (>/=+1 D) was present in 6.0% of the total population, and astigmatism (>/=-1.5 D) was present in 9.5%. CONCLUSIONS: The prevalence of myopia among 12- to 13-year-old children in Mexico is high. The majority of cases are low grade, and a large number of the myopic children do not have, or do not use, prescription glasses. PMID- 12771663 TI - Continuous ambient lighting and lens compensation in infant monkeys. AB - PURPOSE: Protracted daily lighting cycles do not promote abnormal ocular enlargement in infant monkeys as they do in a variety of avian species. However, observations in humans suggest that ambient lighting at night may reduce the efficiency of the emmetropization process in primates. To test this idea, we investigated the ability of infant monkeys reared with continuous light to compensate for optically imposed changes in refractive error. METHODS: Beginning at about 3 weeks of age, a hyperopic or myopic anisometropia was imposed on 12 infant rhesus monkeys by securing either a -3 D or +3 D lenses in front of one eye and a zero-powered lens in front of the fellow eye. Six of these monkeys were reared with the normal vivarium lights on continuously, whereas the other six lens-reared monkeys were maintained on a 12-h-light/12-h-dark lighting cycle. The ocular effects of the lens-rearing procedures were assessed periodically during the treatment period by cycloplegic retinoscopy, keratometry, and A-scan ultrasonography. RESULTS: Five of six animals in each of the lighting groups demonstrated clear evidence for compensating anisometropic growth. In both lighting groups, eyes that experienced optically imposed hyperopic defocus (-3 D lenses) exhibited faster axial growth rates and became more myopic than their fellow eyes. In contrast, eyes treated with +3 D lenses showed relatively slower axial growth rates and developed more hyperopic refractive errors. The average amount of compensating anisometropia (continuous light, 1.6 +/- 0.5 D vs. control, 2.3 +/- 0.5 D), the structural basis for the refractive errors, and the ability to recover from the induced refractive errors were also not altered by continuous light exposure. CONCLUSION: Ambient lighting at night does not appear to overtly compromise the functional integrity of the vision-dependent mechanisms that regulate emmetropization in higher primates. PMID- 12771664 TI - Dynamic accommodative changes in rhesus monkey eyes assessed with A-scan ultrasound biometry. AB - PURPOSE: Prior studies in humans measured time constants of biometric accommodative changes as a function of amplitude, and prior studies in monkeys used slit lamp videography to analyze dynamic lenticular accommodative movements. Neither of these studies related biometric changes to refractive changes. We wished to develop and test methodology to begin to test the hypothesis that ocular biometric changes are well correlated with accommodative refractive changes in rhesus monkeys. METHODS: Methodology is described to dynamically measure biometric accommodative changes with A-scan ultrasonography. Lens thickness, anterior chamber depth, and anterior segment length (anterior chamber depth plus lens thickness) were measured dynamically during Edinger-Westphal stimulated accommodation in two eyes of one rhesus monkey. In addition, dynamic accommodative refractive changes were measured with infrared photorefraction. Functions were fit to the accommodative and disaccommodative responses to obtain time constants. Derivatives of these functions allow peak velocities to be determined for each amplitude. Dynamic changes in lens thickness and anterior chamber depth measured with A-scan biometry were compared with dynamic measures of accommodation using infrared photorefraction. RESULTS: Lens thickness and anterior segment length increase and anterior chamber depth decreases during accommodation. The biometric changes are well correlated with the accommodative optical changes. Peak velocities of accommodative changes in lens thickness and anterior chamber depth increase with amplitude and peak velocities for disaccommodation were higher than those for accommodation. CONCLUSIONS: Dynamic A scan provides a method for dynamic analysis of the accommodative biometric changes during Edinger-Westphal-stimulated accommodation in monkeys, although the measurement resolution of this approach is limited. PMID- 12771665 TI - Stray light of spectacle lenses compared with stray light in the eye. AB - Stray light in spectacle lenses may affect the overall vision. It may also affect the measurement of ocular stray light, contrast sensitivity, or glare sensitivity. This article describes common stray light characteristics for glass and plastic spectacle lenses and compares this to the stray light characteristics of the eye, which are well known from the literature. Stray light is described by the skirts of the point-spread function (PSF), which were measured for angles from 4 degrees to 30 degrees. The PSF of spectacle lenses appears to follow the equation PSF = a(10). (theta/10)b, with a(10) and b representing fitting parameters and theta representing the stray light angle. The slope b is on average -2, which similar to that of the eye. For clean spectacle lenses, the PSF is usually at least an order of magnitude lower than that of the eye, whereas "as worn" (uncleaned) spectacle lenses may approach the PSF of the eye. To reach the PSF of the eye, the spectacle lens needs to be contaminated by as much as one or two fingerprints. The article also shows that plastic spectacle lenses degrade much faster than glass spectacle lenses when looking at the amount of stray light. PMID- 12771667 TI - Cardiorespiratory interactions and blood flow generation during cardiac arrest and other states of low blood flow. AB - PURPOSE OF REVIEW: Recent advances in cardiopulmonary resuscitation have shed light on the importance of cardiorespiratory interactions during shock and cardiac arrest. This review focuses on recently published studies that evaluate factors that determine preload during chest compression, methods that can augment preload, and the detrimental effects of hyperventilation and interrupting chest compressions. RECENT FINDINGS: Refilling of the ventricles, so-called ventricular preload, is diminished during cardiovascular collapse and resuscitation from cardiac arrest. In light of the potential detrimental effects and challenges of large-volume fluid resuscitations, other methods have increasing importance. During cardiac arrest, active decompression of the chest and impedance of inspiratory airflow during the recoil of the chest work by increasing negative intrathoracic pressure and, hence, increase refilling of the ventricles and increase cardiac preload, with improvement in survival. Conversely, increased frequency of ventilation has detrimental effects on coronary perfusion pressure and survival rates in cardiac arrest and severe shock. Prolonged interruption of chest compressions for delivering single-rescuer ventilation or analyzing rhythm before shock delivery is associated with decreased survival rate. SUMMARY: Cardiorespiratory interactions are of profound importance in states of cardiovascular collapse in which increased negative intrathoracic pressure during decompression of the chest has a favorable effect and increased intrathoracic pressure with ventilation has a detrimental effect on survival rate. PMID- 12771668 TI - Strategies for reversing shock-resistant ventricular fibrillation. AB - PURPOSE OF REVIEW: Shock-resistant ventricular fibrillation is defined as ventricular fibrillation persisting after three defibrillation attempts. In approximately 10 to 25% of all cardiac arrests, shock-resistant ventricular fibrillation develops, and 87 to 98% of these patients die. RECENT FINDINGS: In the treatment of shock-resistant ventricular fibrillation, defibrillation using biphasic waveforms is considered as an intervention of choice. Intravenous amiodarone is also acceptable, safe, and useful, based on evidence from two randomized clinical trials. Intravenous vasopressin is acceptable and probably safe and useful, but the evidence supporting this recommendation is coming from a small, randomized clinical trial. Procainamide is acceptable but not recommended. In the presence of acute myocardial infarction and recurrent ventricular fibrillation, if all other therapies fail, beta-blockers can be considered. Magnesium, lidocaine, and bretylium are not recommended in the treatment of shock resistant ventricular fibrillation. SUMMARY: Biphasic defibrillation and intravenous amiodarone are useful in shock-resistant ventricular fibrillation. PMID- 12771669 TI - Preconditioning and the oxidants of sudden death. AB - Sudden cardiac death remains a daunting medical challenge. Rescuers have minutes to defibrillate the heart and prevent ischemic injury to critical organs. Cardiopulmonary resuscitation can extend the window for successful therapy but not for long. Complicating this further is the fact that few new therapies have been proven to protect against the postresuscitation phase of cardiac arrest, when as many as 90% of patients die despite successful defibrillation. Oxidants (both reactive oxygen and nitrogen) likely play critical roles during cardiac arrest, affecting defibrillation success by affecting cardiac gap junctions and after successful defibrillation causing multiorgan damage via direct and programmed cell death. Preconditioning is an intrinsic adaptive response to stress that targets this sequence of events and is highly protective against ischemia/reperfusion injury in the heart, brain, and other critical organs. Thus, how oxidants are affected by preconditioning could provide new insights and therapies for improving both defibrillation success and oxidant-mediated postresuscitation injury of sudden cardiac death. PMID- 12771670 TI - Myocardial protection during resuscitation from cardiac arrest. AB - PURPOSE OF REVIEW: Successful treatment of cardiac arrest requires that an electrically stable and mechanically competent cardiac activity be promptly reestablished. However, many interventions used to attempt to reestablish cardiac activity may also inflict additional myocardial injury and, in turn, compromise resuscitability. In this review, we examine mechanisms of such myocardial injury and discuss potential new strategies for myocardial protection during resuscitation from cardiac arrest. RECENT FINDINGS: Efforts are currently directed at understanding underlying mechanisms of myocardial injury associated with current resuscitation methods, with the purpose of developing alternative approaches that are safer and more effective. These new approaches include, among others, the development of alternative low-energy defibrillation waveforms, methods for optimizing the timing for attempting defibrillation, and the use of vasopressor agents devoid of beta-agonist effects. There is also interest in understanding the role that activation of pathways of ischemic and reperfusion injury could play during resuscitation from cardiac arrest. To this end, activation of the sarcolemmal sodium-hydrogen exchanger isoform 1 seems to play an important role. Other potentially important pathways involve adenosine metabolism, activation of potassium ATP channels, and generation of oxygen radical species. These pathways may become novel pharmacologic targets for cardiac resuscitation. SUMMARY: The growing body of research in these areas is bringing hope that in a not so distant future new approaches and interventions for cardiac resuscitation could be available for resuscitation of humans in various clinical settings. PMID- 12771671 TI - Hypothermia after cardiac arrest: a treatment that works. AB - PURPOSE OF REVIEW: Sudden death from cardiac arrest is a major health problem that still receives too little publicity. Current therapy after cardiac arrest concentrates on resuscitation efforts because, until now, no specific therapy for brain protection after restoration of spontaneous circulation was available. Therapeutic mild or moderate resuscitative hypothermia is a novel therapy with multifaceted chemical and physical effects by preventing or mitigating the derangements seen in the postresuscitation syndrome. RECENT FINDINGS AND SUMMARY: In 2002, two prospective, randomized studies reported improved outcomes when deliberate hypothermia was induced in comatose survivors after resuscitation from cardiac arrest. However, several issues with regard to resuscitative cooling are still unanswered and should be studied further. These include the optimal timing to initiate cooling, the optimal cooling period, the optimal temperature level, and rewarming strategy. Even important questions, such as which cooling technique will be available in the near future that would combine ease of use with high efficacy, are not answered yet. PMID- 12771672 TI - Monitoring during cardiac arrest: are we there yet? AB - Advancements in electronic data acquisition have translated into improved monitoring of victims of cardiac arrest, but initial techniques remain direct observation of pulses and respirations. The most essential monitor continues to be the electrocardiogram. However, monitoring diastolic blood pressure, myocardial perfusion pressure, and end-tidal carbon dioxide are extremely useful. Most of the current research on monitoring during cardiopulmonary resuscitation focuses on methods for analyzing ventricular fibrillation waveforms. By analyzing the waveform, defibrillation shocks may be delivered at the time when the chance of success is optimal. Low-amplitude and low-frequency fibrillation waveforms are associated with increased rates of asystole and pulseless electrical activity after defibrillation. Methods of analyzing the ventricular fibrillation waveform include measuring the amplitude and frequency and combining the contributions of amplitude and frequency by various methods to improve discrimination. Other types of monitoring being studied for their usefulness during cardiac arrests include sonography, Bispectral Index monitoring, tissue carbon dioxide monitors, and pupil observation. The test of these monitoring techniques is ultimately their ability to improve patient survival to hospital discharge, which is a major challenge for resuscitation researchers. PMID- 12771673 TI - Cardiopulmonary Resuscitation Guidelines 2000 update: what's happened since? AB - PURPOSE OF REVIEW: To examine the literature for new resuscitation science since the publication of the Guidelines 2000 for Cardiopulmonary Resuscitation and Emergency Cardiac Care. RECENT FINDINGS: The two and a half years since the publication of the Guidelines 2000 have seen the advent of a number of new and important resuscitation studies. Such studies highlight the importance of simplification of cardiopulmonary resuscitation techniques and guidelines, including the elimination of the layperson pulse check and the need for a simple form of basic life support cardiopulmonary resuscitation that decreases interruptions of chest compressions. Automatic external defibrillators, even in the hands of nontraditional first responders, are effective and safe. A second prospective, randomized clinical trial of amiodarone for refractory ventricular fibrillation has again shown positive results in improving survival to hospital admission. Finally, mild hypothermia appears to be the first effective therapy at decreasing central nervous system injury when administered after resuscitation. SUMMARY: In this report, we review these new studies and discuss how they corroborate or alter the published 2000 guidelines. PMID- 12771674 TI - Rationale for cardiovascular monitoring. AB - A primary aspect of cardiovascular support of the critically ill patient is the titration of cardiopulmonary therapies based on the baseline cardiopulmonary status and the subsequent physiologic response. Implicit in this paradigm is the monitoring of the processes being titrated. The degree to which a specific physiologic variable, such as mean arterial pressure or arterial oxygen saturation, needs to be assessed is a function of the therapy used, the stability of the patient, the relation among the variables defining the hemodynamic profile, and the ability of the support staff to remain in close attendance at the bedside. In an otherwise stable patient in heart failure being treated with mild afterload reduction and diuretics, periodic measures of heart rate, urine output, and daily measures of body weight are all that are reasonably needed to titrate therapy. However, in the management of a patient with cardiogenic shock with pulmonary edema and respiratory failure, continuous measures of mean arterial pressure, left ventricular filling pressure, cardiac output, arterial oxygen content, and end-organ function may be necessary as more potent and risky therapies are used. How, then, does one arrive at the correct formula to prescribe appropriate physiologic monitoring for the patient in the intensive care unit setting? To a large extent this is unknown, primarily because the utility of monitoring techniques to diagnose pathophysiologic processes and the resultant effect of therapy to reverse it is not known for most of the diseases treated in the intensive care unit. Few monitoring techniques have progressed through a logical progression of development to their present level of use. Thus, their use in the management of the critically ill patient cannot be vigorously defended, except under specific conditions. PMID- 12771675 TI - Cardiovascular monitoring tools: use and misuse. AB - PURPOSE OF REVIEW: To review important areas of current and novel hemodynamic monitoring practice in the intensive care unit and to highlight potential areas of physiologic and clinical use or misuse, as well as areas of uncertainty and ongoing controversy. RECENT FINDINGS: To truly determine when hemodynamic monitoring tools are misused would require randomized controlled evidence of a measurable improvement in relevant clinical (as opposed to physiologic) outcomes. Unfortunately, little evidence of this kind exists, and that which does exist is highly controversial in nature. Because of the limited evidence of an effect of hemodynamic monitoring on clinical outcomes, the use and misuse of hemodynamic monitoring tools is typically judged on physiologic grounds (Does it improve physiology? Does it predict physiology? Is it physiologically rational?). The relation between physiologic gain and final clinical outcome, however, is tenuous. Recent investigations confirm this lack of a clear link. They also suggest that new technology that is now emerging to less invasively measure cardiac output and intrathoracic fluid compartments is ready for formal evaluations of efficacy and effectiveness. SUMMARY: The effectiveness of hemodynamic monitoring in the intensive care unit remains inadequately tested and unproven. New tools are now rapidly emerging to challenge established technologies. Formal assessment of their efficacy and effectiveness is needed to avoid a repeat of the pulmonary artery catheter experience. PMID- 12771676 TI - Respiratory monitoring tools in the intensive care unit. AB - PURPOSE OF REVIEW: Respiratory monitoring tools remain important in the overall assessment of a patient in the ICU. A working knowledge of the various tools is helpful in applying their use to patient care. RECENT FINDINGS: The past year has seen tremendous research in this domain of patient care, fueled by rapid advances in modern technology. Still, the ICU clinician continues to rely on tried and true established tools while awaiting proof of clinical efficacy of newer modalities. SUMMARY: This review discusses traditional methods of respiratory system evaluation and their current controversies. Attention is also given to newer modalities, including those that are investigational or currently limited to bench application, that show promise of future application in ICU clinical practice. PMID- 12771677 TI - Treatment algorithms and protocolized care. AB - PURPOSE OF REVIEW: Excess information in complex ICU environments exceeds human decision-making limits and likely contributes to unnecessary variation in clinical care, increasing the likelihood of clinical errors. I reviewed recent critical care clinical trials searching for information about the impact of protocol use on clinically pertinent outcomes. RECENT FINDINGS: Several recently published clinical trials illustrate the importance of distinguishing efficacy and effectiveness trials. One of these trials illustrates the danger of conducting effectiveness trials before the efficacy of an intervention is established. The trials also illustrate the importance of distinguishing guidelines and inadequately explicit protocols from adequately explicit protocols. Only adequately explicit protocols contain enough detail to lead different clinicians to the same decision when faced with the same clinical scenario. SUMMARY: Differences between guidelines and protocols are important. Guidelines lack detail and provide general guidance that requires clinicians to fill in many gaps. Computerized or paper-based protocols are detailed and, when used for complex clinical ICU problems, can generate patient-specific, evidence based therapy instructions that can be carried out by different clinicians with almost no interclinician variability. Individualization of patient therapy can be preserved by these protocols when they are driven by individual patient data. Explicit decision-support tools (eg, guidelines and protocols) have favorable effects on clinician and patient outcomes and can reduce the variation in clinical practice. Guidelines and protocols that aid ICU decision makers should be more widely distributed. PMID- 12771678 TI - Powerful partnerships achieve nursing's greatest outcomes. PMID- 12771679 TI - Embracing "the 'A' word". PMID- 12771681 TI - Medication management update. AB - To ensure compliance with the Joint Commission's newest medication standards, review the following details. PMID- 12771682 TI - Strategies at work: simple steps to satisfaction. PMID- 12771683 TI - Overcome hidden expenses, migrating staff. PMID- 12771684 TI - How heart failure complicates care. AB - Review why heart failure (HF) develops and how it affects the nursing care of patients with other health problems. PMID- 12771685 TI - Bring barcoding to the bedside. AB - A 240-bed regional hospital shares best practices for implementing a patient safety initiative that targets point-of-care barcode technology. PMID- 12771687 TI - Shining through a merger. AB - Two Nebraska hospitals that merged offer staff retreats to help rebuild trust and the organization's nursing identity. PMID- 12771686 TI - Peer-to-peer education: nighttime is the right time. AB - The Atlanta VA Hospital created a peer-oriented committee to advance the continuing education of its nonday-shift personnel. PMID- 12771688 TI - Nursing burn injuries. AB - Explore a cross-section of current burn injury practices and nurses' roles in these practices. PMID- 12771689 TI - Monitor outcomes for safer infusion therapy, better resource management. AB - To determine how to properly implement new CDC evidence-based practices, collect, analyze, and apply outcomes data to provided infusion care. PMID- 12771691 TI - Is the paper strip printed directly from the electronic fetal monitor still necessary for ongoing fetal heart rate interpretation during labor? PMID- 12771693 TI - Use of upright positioning with epidural analgesia: findings from an observational study. AB - PURPOSE: To present research findings and related nursing implications from an observational study designed to evaluate the use of upright positioning during second stage labor with patients who had received low-dose epidural analgesia. STUDY DESIGN AND METHODS: This descriptive study evaluated outcomes from a sample of 74 healthy women having their first childbirth. They had all received epidural analgesia during the first and second stages of labor. Data were also collected by nurses on the use of birthing beds, and the extent of physical and emotional support the women needed while following the upright positioning study protocol. RESULTS: All women were able to maintain upright positions throughout the second stage of labor following epidural analgesia administration. No adverse neonatal outcomes or maternal problems (such as excessive vaginal bleeding) were documented. CLINICAL IMPLICATIONS: Although women were capable of assuming upright positions during second stage, the study results indicated that constant physical and emotional support was necessary for most women. Future research on methods to prepare women for multiple position options after administration of low-dose epidural analgesia should be undertaken. In addition, nurses should evaluate the benefits of upright positioning in terms of facilitating progress of labor. PMID- 12771694 TI - Maternal management behaviors for young children with type 1 diabetes. AB - PURPOSE: To describe the process that mothers raising young (0-4 years old) children who are newly diagnosed with type 1 diabetes move through to attain the necessary skills to care for their children. STUDY DESIGN AND METHODS: A mixed methods design was used, including qualitative interviews with 28 mothers of young children with type 1 diabetes. Principles of naturalistic inquiry were used to guide the data collection process, management, and analysis of the qualitative findings. RESULTS: The process paralleled two of three management approaches and associated behaviors previously described by Gallo and Knafl. Strict adherence behaviors included rigidly following the team recommendations and avoiding strange environments outside the home. Flexible adherence behaviors strove to bring spontaneity back into family life. Selective adherence was not used by this population. CLINICAL IMPLICATIONS: Nurses working with these mothers can provide information and support to help them transition from using strict adherence to the more user-friendly flexible adherence, while avoiding the pitfalls of the possibly harmful third approach of selective adherence. Nurses need to remember to praise the parents' efforts at managing their children's diabetes, for our acknowledgment of their work is empowering and affirming. PMID- 12771695 TI - Men in obstetrical nursing: perceptions of the role. AB - PURPOSE: To explore the role of men as obstetrical nurses. DESIGN: Exploratory cross sectional survey using structured and open-ended questions. SAMPLE: Three groups: 599 men licensed as Registered Nurses by the Commonwealth of Massachusetts, 337 District I AWHONN members, and 130 pregnant women. ANALYSIS: Univariate, bivariate, and logistic regression were performed for the AWHONN and pregnant women groups. Descriptive and narrative summaries for the men who were Registered Nurses. RESULTS: Seventy-three percent of AWHONN members had positive attitudes toward men in obstetric nursing. The experience of having worked with men in obstetrical nursing roles was the most significant predictor of positive perceptions by AWHONN members. Nurses in the positions of clinical nurse specialists or nurse educators in academia had more negative perceptions of men in this specialty. No predictor variables reached significance for the group of pregnant women. Only 6.8% of the male RNs questioned had ever worked in the specialty, and most reported a lack of interest in working in the specialty of obstetrics. CLINICAL IMPLICATIONS: Findings suggest that both clinical and academic settings may need to adopt more nontraditional recruitment and teaching strategies to encourage men to pursue this specialty. PMID- 12771696 TI - Components of developmental care and the evidence for their use in the NICU. AB - Developmental care, a philosophy of care that requires rethinking the relationships between infants, families, and healthcare providers, is in place in the majority of neonatal intensive care units in the United States. Developmental care includes a variety of activities designed to manage the environment and individualize the care of the premature infant based on behavioral observations. The goal is to promote a stable, well-organized infant who can conserve energy for growth and development. Research about the effects of developmental care has shown a trend toward improved short-term physiologic, development, and resource utilization outcomes for infants up to 24 months of age, but benefits beyond this age are unclear. Most of the research has focused on developmental care as a whole, but there is also strong scientific evidence for specific components of developmental care. The NICU care provider should use developmental care interventions that are clearly supported by evidence, and use others based on judgment and the infant's responses. PMID- 12771697 TI - Pica in pregnancy: does it affect pregnancy outcomes? AB - PURPOSE: To discover the prevalence of pica, the documentation of pica on medical records, and any relationship of pica to pregnancy outcomes in rural socioeconomically disadvantaged pregnant women. STUDY DESIGN AND METHODS: Prospective, descriptive, correlational investigation with 128 women who sought prenatal care from two rural community health agencies. Demographic and sociocultural variables, pica practices, pica substances ingested, and pregnancy outcomes were collected. RESULTS: Thirty-eight percent of these pregnant women practiced pica. African-American women reported practicing pica more often than other ethnicities. Substances ingested included ice (>1 cup/day), freezer frost, laundry starch, cornstarch, clay dirt, and baked clay dirt. Polypica (ingestion of more than one substance) was practiced by 11 women. Women practicing pica were more likely to have been underweight prior to pregnancy, and smoked fewer cigarettes. Women reporting daily pica practice were significantly more likely to have lower prenatal hematocrits than women who did not practice pica, or who practiced pica less frequently than daily. No specific pregnancy complication was associated with the practice of pica. CLINICAL IMPLICATIONS: Pica exists, and might be more common than healthcare providers assume. Although this study did not show specific pregnancy complications associated with pica, other studies have shown anemia and lead poisoning among women who practice pica. It is not clear that patients volunteer information about pica, so it would be helpful if nurses queried patients at each prenatal visit regarding pica practice. Discussion of pica practices should be based on a nonjudgmental model, for pica may have strong cultural implications, and may be practiced for cultural reasons unknown to the nurse. PMID- 12771698 TI - What we can learn from the British maternal child health system. AB - Infants born in Great Britain have a better chance of living until their first birthdays than do infants born in the United States. Although Great Britain spends less than half what the United States spends on healthcare, its estimated infant mortality rate was 5.5 as compared to 6.8 in the United States in 2001 (Central Intelligence Agency [CIA], 2001). What is different about healthcare in Great Britain? This article details some of the differences, suggesting that there are ways the United States can improve its infant mortality rates. PMID- 12771699 TI - Emergency contraception for prevention of adolescent pregnancy. AB - Adolescent pregnancy remains a significant problem in the United States today, despite availability of effective contraceptive methods. Not all sexually active adolescents use contraception, and even those who do use contraception sometimes use it incorrectly. Emergency contraception, which refers to methods of pregnancy prevention used after unprotected intercourse, has the potential to prevent most unplanned adolescent pregnancies. Emergency contraceptive pills (ECP) containing estrogen and progestin or progestin alone are more than 75% effective when the first dose is taken within 72 hours after unprotected sex and the second dose is taken 12 hours later. However, barriers to accessing ECPs include lack of knowledge of the method, fear of loss of privacy, difficulties in finding a provider, and cost. Another barrier is that controversy exists about the mechanisms of action of emergency contraception about its role in pregnancy prevention. As a result, some nurses are not comfortable with suggesting emergency contraception to their patients. Nurses can play a critical role in providing ECPs to adolescents by developing programs to streamline distribution of ECPs, while maintaining adolescent privacy. Other essential roles for nurses include providing education about ECPs to parents, other healthcare providers and community members, and advocating for political and legal changes that will ease restrictions on ECP distribution. Nurses who are personally uncomfortable discussing emergency contraception can refer their patients to other providers for information and access to this method. PMID- 12771700 TI - Adolescent girls' personal experience with Baby Think It Over infant simulator. AB - PURPOSE: To explore adolescent girls' personal experience with an infant simulator that had to be cared for over a period of 1 to 2 weeks. STUDY DESIGN AND METHODS: This qualitative study employed the phenomenological approach and utilized Colaizzi's method of analysis. Participants were nine adolescent high school girls who were interviewed and audiotaped. Interview data were coded using NUD*IST 4 software. RESULTS: Three themes emerged from the data. They were (1) a parenting journey incorporating intellectual, emotive, and physical faculties; (2) recognizing the illusionary nature of previously held ideas about parenting an infant; and (3) offering counsel based on the BTIO experience. CLINICAL IMPLICATIONS: The new perspectives gained by the participants about parenting a newborn infant differed markedly from the romantic fantasies they held prior to the experience. The teens started thinking more seriously about the consequences of sexual activity. Used for 1 to 2 weeks, in conjunction with budgeting exercises and provocative small-group discussions, the BTIO shows promise as an effective tool in helping to modify unrealistic and idealized teen views of adolescent pregnancy and parenting. PMID- 12771701 TI - How nurses can help women whose spouses are deployed to a war zone. PMID- 12771702 TI - Tumor markers in the diagnosis of primary bladder cancer. A systematic review. AB - PURPOSE: We systematically reviewed the available evidence, and obtained and compared summary estimates of the sensitivity and specificity of cytology and the urine based markers bladder tumor antigen, BTA stat (Polymedco, Redmond, Washington), BTA TRAK (Polymedco), NMP22 (Matritech, Cambridge, Massachusetts), telomerase and fibrin degradation product in detecting primary bladder cancer. MATERIALS AND METHODS: Studies on the diagnosis of primary bladder cancer published from 1990 through November 2001 in English and German were retrieved from MEDLINE and EMBASE data bases. In our research we included studies that evaluated 1 or more of the markers, used cystoscopy as the reference standard and allowed the construction of a 2 x 2 contingency table for a per patient analysis. The data plus items on study and clinical characteristics were extracted by 2 observers. Sensitivity and specificity for each marker were estimated using a bivariate random effect meta-analysis. A multivariable analysis was performed to explain study variation. RESULTS: A total of 42 studies were included in our review. Only 2 studies were available on fibrin degradation product, hence a meta analysis was not possible. Cytology had the best specificity at 94% (95% CI: 90% to 96%). This figure was significantly better than that of the other markers except for telomerase (specificity 86% [71% to 94%]). Telomerase had the best sensitivity (75% [71% to 79%]) but it was not significantly better than that of BTA stat (70% [66% to 74%]). Case control designs yielded lower values for sensitivity for the tumor markers cytology, bladder tumor antigen and BTA stat. CONCLUSIONS: Cytology has the best specificity and telomerase the best sensitivity. However, none of the markers studied here is sensitive enough to be recommended for daily routine. PMID- 12771703 TI - Improving and predicting radiosensitivity in muscle invasive bladder cancer. AB - PURPOSE: Muscle invasive bladder cancer is a common urological malignancy with a relatively poor prognosis and 5-year survival rates ranging from 20% to 90%. We review methods of improving the outcome of this condition, with particular emphasis on the principal bladder preserving treatment modality of radiation therapy. MATERIALS AND METHODS: We performed a literature search using MEDLINE and the ISI Web of Science using the keywords radiotherapy, radiosensitization and bladder neoplasia to ascertain the current status of radiation therapy and radiosensitizing agents in the treatment of muscle invasive bladder cancer. RESULTS: Several methods aimed at improving outcome following radiation therapy for muscle invasive bladder cancer are described. These methods range from modifications in the application of radiation therapy to use of conventional radiosensitizing agents, such as accelerated radiotherapy with carbon dioxide, oxygen and nicotinamide, and finally to use of more novel agents that interact with oncogenic products. The use of assays that predict tumor sensitivity on an individual basis represents an additional potential method to improve prognosis following radiation therapy. CONCLUSIONS: The ability to predict tumor radiosensitivity and the subsequent implementation of radiosensitizing techniques are likely to improve the results of treatment centered on radiation therapy, suggesting that bladder sparing approaches will remain a treatment option for muscle invasive bladder cancer. PMID- 12771704 TI - The management of high risk prostate cancer. AB - PURPOSE: A major problem that urologists encounter is the recurrence of prostate cancer after local therapy or the failure of primary therapy. There is no consensus on the treatment of these groups of patients. The role of radiotherapy, hormonal therapy or chemotherapy must be defined. MATERIALS AND METHODS: A comprehensive literature review of the current management of recurrence after primary therapy in prostate cancer was performed using MEDLINE, a review of current urology and oncology journals, and abstracts from recent urology meetings. The data collected focused on the role of radiotherapy, hormonal therapy and chemotherapy in this setting. RESULTS: Defining a high risk group of patients using Gleason score, seminal vesicle or pelvic lymph node involvement and prostate specific antigen recurrence time is important for treating the problem early in disease course. Adjuvant radiotherapy cannot offer a survival advantage but it provides longer biochemical recurrence-free survival. Early administration of salvage radiotherapy in adequate doses provides a success rate similar to that of adjuvant radiotherapy in patients with low prostate specific antigen. Although there is no good evidence that early androgen deprivation definitely prolongs patient survival, increased time to progression in addition to excellent palliation with early hormonal therapy was reported in the majority of trials. Early trials of the use of chemotherapy showed improved survival rates with adjuvant chemotherapy alone or with a combination of hormonal therapy, especially in patients with nonmetastatic disease. CONCLUSIONS: Although analysis of current literature revealed that patients who have recurrence after primary therapy would benefit from radiotherapy, hormonal therapy, chemotherapy or a combination of therapies, additional prospective randomized studies are needed to support these findings. PMID- 12771705 TI - Erectile dysfunction in the elderly: epidemiology, etiology and approaches to treatment. AB - PURPOSE: Erectile dysfunction is experienced at least some of the time by most men who have reached 45 years of age, and it is projected to affect 322 million men worldwide by 2025. The prevalence of erectile dysfunction is high in men of all ages and increases greatly in the elderly. MATERIALS AND METHODS: This paper reviews the epidemiology of erectile dysfunction with an emphasis on the experience of older men, normal age related changes in the structure and function of the penis that may contribute to increased risk with age, how the accumulation of risk factors with age may contribute to the high prevalence of the disease in older men, and established and emerging therapies. The normal aging process and age related risk factor accumulation contribute to the increased prevalence of erectile dysfunction in the elderly. RESULTS: Remarkable progress has been made in the treatment of erectile dysfunction. At present inhibition of phosphodiesterase 5 with oral agents such as sildenafil would appear to be the initial treatment of choice. These drugs have been shown to be safe and effective, and sildenafil has demonstrated efficacy in patients with many of the comorbidities observed in older men with erectile dysfunction. New treatments, in particular transfection with genes for key mediators of erectile function that are known to be down-regulated in elderly men, also hold promise. CONCLUSIONS: Further research into the neural, vascular and molecular mechanisms involved in penile erection will lead to the development of even safer, more effective and more convenient therapies for men with erectile dysfunction. PMID- 12771706 TI - Randomized controlled trial of zoledronic acid to prevent bone loss in men receiving androgen deprivation therapy for nonmetastatic prostate cancer. AB - PURPOSE: A multicenter double-blind, randomized, placebo controlled clinical trial was performed to assess the effect of zoledronic acid, a potent new bisphosphonate, on bone mineral density during androgen deprivation therapy for nonmetastatic prostate cancer. MATERIALS AND METHODS: Men with M0 (no distant metastases) prostate cancer beginning androgen deprivation therapy were randomly assigned to receive 4 mg. zoledronic acid or placebo intravenously every 3 months for 1 year. The primary efficacy variable was the percent change from baseline to 1 year in bone mineral density of the lumbar spine as measured by dual energy x ray absorptiometry. RESULTS: A total of 106 men were enrolled in the trial. Mean bone mineral density in the lumbar spine increased by 5.6% in men receiving zoledronic acid and decreased by 2.2% in those given placebo (mean difference 7.8%, 95% confidence interval 5.6%-10.0%, p <0.001). Mean bone mineral density of the femoral neck, trochanter and total hip also increased in the zoledronic acid group and decreased in the placebo group. Zoledronic acid was well tolerated. CONCLUSIONS: Zoledronic acid increases bone mineral density in the hip and spine during androgen deprivation therapy for nonmetastatic prostate cancer. PMID- 12771707 TI - Multiple arteries in live donor renal transplantation: surgical aspects and outcomes. AB - PURPOSE: This retrospective study describes the surgical techniques and outcomes of live donor renal allografts with multiple arteries. MATERIALS AND METHODS: Between 1976 and 2000, 1,200 consecutive live donor renal transplants were done, including 1,087 with single (group 1) and 113 with multiple (group 2) arteries. Intracorporeal in situ anastomotic techniques were used for 94 grafts with multiple arteries, while ex vivo techniques were used for 19. During in situ surgery each one of the multiple arteries was anastomosed separately to an individual artery. In ex vivo surgery 2 or more arteries were joined together on the bench to form a common stem, which was then anastomosed to an iliac artery or the aorta. RESULTS: Patient and graft survival were comparable in groups 1 and 2. The 2 groups were comparable regarding complications, including arterial bleeding, hematoma, renal artery stenosis, acute rejection, new onset hypertension, acute tubular necrosis and urological complications. Mean serum creatinine +/- SD at 1 year was 1.4 +/- 0.5 and 1.5 +/- 0.6 mg./dl., and at 5 years it was 1.8 +/- 1 and 2.1 +/- 1.4 mg./dl. for the 2 groups, respectively. The difference was only significant at 1 year (p = 0.02). Graft and patient survival, and the incidence of the described complications were comparable for the ex vivo bench anastomotic techniques and intracorporeal in situ techniques in the group with multiple renal arteries. CONCLUSIONS: The use of multiple arteries in renal allografts does not adversely affect patient or graft survival. It is not associated with an increased rate of complications except for significantly higher mean serum creatinine at 1 year. Extracorporeal bench surgery was as effective as intracorporeal surgery for the anastomosis of multiple renal arteries with no increase in the incidence of relevant complications. PMID- 12771708 TI - Quality of life, pain and return to normal activities following laparoscopic donor nephrectomy versus open mini-incision donor nephrectomy. AB - PURPOSE: We evaluated pain, convalescence and health related quality of life in patients undergoing laparoscopic and open mini-incision donor nephrectomy. MATERIALS AND METHODS: We compared the records of consecutive patients who underwent laparoscopic and mini-incision open donor nephrectomy from our donor nephrectomy data base in retrospective fashion using 2 questionnaires. The first questionnaire evaluated postoperative pain, return to functioning time and satisfaction. The second questionnaire was the RAND 36-Item Health Survey, version 2, a standardized and validated health survey quality of life assessment tool. Mean patient sex, age and followup were similar for the 2 groups. All data were analyzed using the 2-tailed t test for independent variables with commercially available statistical analysis software. RESULTS: Pain in the laparoscopic group was significantly less than in the mini-incision group at all followup time points (p <0.05). Statistically significant differences demonstrated that laparoscopy led to more rapid recovery time in certain categories, including walking, discontinuation of prescribed oral pain relievers, return to driving, and resumption of normal work and home daily activities. More subjective questions in the survey showed high levels of acceptance for the 2 procedures. Using the RAND 36-Item Health Survey, version 2 health related quality of life was significantly higher in the laparoscopy group in 3 domains that measure bodily pain, physical functioning and emotional role functioning. However, each group scored at or above age matched American averages in all domains. CONCLUSIONS: The laparoscopy group had significantly less postoperative pain and required less time to return to normal functional activities than the mini-incision group. In addition, the laparoscopic group showed significantly higher quality of life scores than the mini-incision group in 3 domains. PMID- 12771709 TI - Management of lymphoceles after renal transplantation: laparoscopic versus open drainage. AB - PURPOSE: Laparoscopic surgery has become widely accepted for the treatment of lymphoceles following kidney transplantation. In this single center study we retrospectively reviewed our results of the surgical management of post transplant lymphoceles, assessing indication and outcome of laparoscopic versus open drainage. MATERIALS AND METHODS: The records of 60 patients who underwent surgical treatment for a symptomatic lymphocele following kidney transplantation or combined kidney/pancreas transplantation were retrospectively reviewed. RESULTS: Between 1995 and 2002, 1,836 patients received a kidney transplant at the University of California San Francisco. In 60 patients (3.3%) a symptomatic lymphocele developed and either laparoscopic (20) or open drainage (40) was completed. The conversion rate from laparoscopic to open drainage was 16.5%. The most common indications for open lymphocele drainage were noninfectious wound complications (13 patients) and a high risk of vessel or ureter injury (8) due to proximity of the lymphocele to hilar structures. Additional surgery on the graft was required in 5 patients. Intraoperative blood loss was significantly lower in the laparoscopy group. Median hospital stay was 1 day in the laparoscopy group versus 4 days in the open drainage group. No perioperative complications were observed in either group. After a median followup of 38 months, 2 patients in each treatment group had a symptomatic recurrence. CONCLUSIONS: Although both surgical approaches are safe and effective, laparoscopic drainage should remain the method of choice for the treatment of post-transplant lymphocele. However, open drainage should be performed in patients with wound complications and in those with a small lymphocele adjacent to vital renal structures. PMID- 12771710 TI - Conversion of calcium oxalate to calcium phosphate with recurrent stone episodes. AB - PURPOSE: We have extended our previous observation that the percent occurrence of calcium oxalate stones decreased while that of calcium phosphate stones increased with each new stone event. MATERIALS AND METHODS: The National VA Crystal Identification Center has analyzed veteran patient urinary tract stones from VA hospitals throughout the United States since 1983. We reviewed the composition of 33,198 stones with emphasis on the changes in composition. More than 11,786 stones came from 5,088 recurrent stone formers. Stones were analyzed using high resolution x-ray powder diffraction and Fourier transform infrared spectroscopic techniques. When the stones were investigated as a function of time, it was determined that there was greater variability when samples were more than 30 days apart. RESULTS: The percent occurrence of whewellite, weddelite, apatite, brushite and uric acid in stones increased between 1.0% and 5.9% since our previous study. The percent occurrence of struvite decreased by 2.6%. The percent of calcium oxalate stones decreased while that of calcium phosphate stones increased with each new event. However, the total percent occurrence of all calcium containing stones did not significantly change with recurrent stone events. CONCLUSIONS: Our study suggests a strong trend for the conversion of stone disease from calcium oxalate to calcium phosphate containing stones, which could influence the progression and severity of disease. PMID- 12771711 TI - The urinary response to an oral oxalate load in recurrent calcium stone formers. AB - PURPOSE: Dietary oxalate may contribute up to 50% to 80% of the oxalate excreted in urine. We studied the urinary response to an oral oxalate load in male and female idiopathic recurrent calcium oxalate stone formers with and without mild hyperoxaluria to evaluate the potential pathophysiological significance of dietary oxalate. MATERIALS AND METHODS: A total of 60 recurrent calcium stone formers underwent an oral oxalate load test. Urine samples were obtained after an overnight fast. Each patient then received an oral oxalate load (5 mM. sodium oxalate dissolved in 250 ml. distilled water) and 3, 2-hour urine samples were obtained 2, 4 and 6 hours after the oxalate load. We compared the response to the oxalate load in patients with and without mild hyperoxaluria, and in male and female patients without hyperoxaluria. RESULTS: The peak urinary response occurred 4 hours after the oral oxalate load in all patients. Those with mild hyperoxaluria had a mean fasting urinary oxalate-to-creatinine ratio +/- SE of 0.027 +/- 0.003 and a mean peak urinary oxalate-to-creatinine ratio of 0.071 +/- 0.006. In comparison, patients with normal oxalate excretion had a fasting and peak urinary oxalate-to-creatinine ratio of 0.018 +/- 0.001 and 0.056 +/- 0.004, respectively (p <0.05). The mean 6-hour increment for urinary oxalate excretion after the oxalate load for patients with hyperoxaluria versus those with normal urinary oxalate excretion was 17.2 +/- 1.9 versus 12.1 +/- 0.98 mg. (p <0.05). In the subset of patients with normal urinary oxalate excretion mean 6-hour cumulative urinary oxalate excretion was 16.8 +/- 1.3 and 13.3 +/- 1.4 mg. in males and females, respectively (p not significant). CONCLUSIONS: Recurrent calcium stone formers with mild hyperoxaluria have higher fasting urinary oxalate and an exaggerated urinary response to an oral oxalate load compared with recurrent calcium stone formers with normal urinary oxalate excretion. Men and women stone formers without hyperoxaluria excrete similar fractions of an oral oxalate load. Increased gastrointestinal absorption and renal excretion of dietary oxalate may be a significant pathophysiological mechanism of stone formation in patients with mild hyperoxaluria. PMID- 12771712 TI - Immediate closure of nephrostomy tube wounds using a tissue adhesive: a novel approach following percutaneous endourological procedures. AB - PURPOSE: We assessed the feasibility of immediate sealing of nephrostomy tube wounds after percutaneous surgery using a tissue adhesive. MATERIALS AND METHODS: The study represents a prospective series of 27 consecutive percutaneous procedures. After nephrostographic exclusion of infrarenal urinary obstruction the nephrostomy tubes were removed and the wound edges were glued together using 2-octyl cyanoacrylate. The wound was covered by gauze to assess the efficiency of sealing and the patients were followed clinically. Another consecutive series of 20 patients who had been treated during 6 months before the current study were used for comparison. The nephrostomy wound in this group was dressed and left to close spontaneously. RESULTS: A total of 27 percutaneous procedures were performed in 25 patients with a median age of 51 years (range 9 to 77). There were 26 cases of percutaneous nephrolithotomy for an average stone burden of 32.6 mm. (range 16 to 70) and 1 pediatric case of percutaneous antegrade balloon dilation of ureteral stricture related to Cohen reimplantation. Median size of the nephrostomy tubes was 16Fr (range 12Fr to 24Fr) and they were maintained a median of 4 days (range 1 to 16) postoperatively. Urinary leakage ceased immediately after tissue adhesive application in all cases. One patient in whom renal colic developed secondary to edema of the ureteral orifice underwent temporary stenting in retrograde fashion. There were no additional complications at a median followup of 5 months (range 3 to 7). The study group had a significantly shorter hospital stay than the wound dressing group (p <0.001). CONCLUSIONS: Wound sealing following nephrostomy tube removal using 2-octyl cyanoacrylate appears to be a safe, simple and efficient method for immediate abolishment of urinary leakage. This novel approach avoids patient and medical personnel inconvenience, permitting early release from the hospital without physical and social limitations related to persistent wound urinary discharge. PMID- 12771713 TI - Laparoscopic pyeloplasty for secondary ureteropelvic junction obstruction. AB - PURPOSE: Laparoscopic pyeloplasty has become a viable option for the treatment of select patients with primary ureteropelvic junction obstruction with success rates similar to those of open surgery. However, little has been written on the application of this technique for secondary ureteropelvic junction obstruction. We report the largest series of secondary ureteropelvic junction obstruction managed by laparoscopic pyeloplasty. MATERIALS AND METHODS: Between March 1994 and March 2001, 36 patients underwent laparoscopic transperitoneal pyeloplasty for secondary ureteropelvic junction obstruction. The patients had undergone an average of 1.3 ureteropelvic junction procedures (range 1 to 4) prior to presentation, including cutting balloon retrograde endopyelotomy in 28, antegrade endoscopic endopyelotomy in 7, retrograde endoscopic endopyelotomy in 4, retrograde balloon dilation in 4 and open pyeloplasty in 3. A preoperative diagnosis of recurrent obstruction was confirmed by renal scan in 31 cases, retrograde pyelography in 2 and computerized tomography in 3. Of the 31 patients who underwent spiral computerized tomography angiogram 87% had crossing vessels. Laparoscopic repair comprised dismembered pyeloplasty in 31 cases, Fengerplasty in 3 and flap repair in 2. Postoperative renal scan or excretory urography objective followup was available for all patients at a mean of 10 months (range 3 to 40). Postoperative subjective patient well-being was assessed using an analog pain scale at a mean followup of 21.8 months (range 3 to 85). RESULTS: Average operative time was 6.2 hours (range 2.7 to 10). Average hospital stay was 2.9 days (range 1 to 7). One intraoperative complication occurred, that is bleeding necessitating conversion to an open procedure. Postoperative complications occurred in 8 cases, including anastomotic leakage in 4, and urinary tract infection, pneumonia, atelectasis, fever, bilateral upper extremity weakness and stone formation 2 months postoperatively in 1 each. On excretory urography, furosemide renal scan or the Whitaker test 32 of 36 patients (89%) had a widely patent ureteropelvic junction. Two patients (5.5%) had equivocal radiographic studies but were asymptomatic. In 2 patients the ureteropelvic junction was obstructed by renal scan. One patient had an indwelling stent for renal function deterioration and 1 was asymptomatic. Hence, 34 of the 36 patients (94%) had a reasonable objective response. Overall a 50% or greater decrease in pain was seen in 32 of 36 patients (89%). In the 4 patients with a less than 50% decrease in pain objective renal scans showed an open ureteropelvic junction. As such, the overall success rate of a greater than 50% decrease in pain, a patent ureteropelvic junction and stable or improved function of the affected renal unit was 83% (30 of 36 patients). CONCLUSIONS: For secondary ureteropelvic junction obstruction, laparoscopic pyeloplasty can be performed safely with a success rate comparable to that of standard open pyeloplasty. The patient benefits of laparoscopic ureteropelvic junction repair of secondary ureteropelvic junction obstruction are similar to the benefits of laparoscopic repair of primary ureteropelvic junction obstruction. PMID- 12771714 TI - A simplified 5-step model for training laparoscopic urethrovesical anastomosis. AB - PURPOSE: We developed a training model and program aimed at improving the skills of urologists with no previous experience in laparoscopy to perform a running suture urethrovesical anastomosis as is done during laparoscopic radical prostatectomy. MATERIALS AND METHODS: Our program is performed on a pelvic trainer with a videolaparoscopic unit and consists of passage of a ligature, intracorporal knotting, intracorporal suturing, linear anastomosis and circular running suture anastomosis. The trainees performed the first 3 tasks during the initial lessons and then advanced to the 2 final tasks. At the end of each lesson time was recorded and progression curve was plotted for each participant for each task. The end point of the study was participant ability to perform an accurate circular anastomosis. Logarithmic regression analysis was used to assess the significance of progression. RESULTS: All 10 urologists who participated in this study showed a rapid and significant decrease in the time required to perform the first 3 tasks accurately. The participants were able to perform a linear anastomosis after 3 to 5 lessons and an accurate circular anastomosis after 5 to 10 lessons. CONCLUSIONS: By using this model and dividing a complicated surgical step to simplified tasks, we were able to improve trainee performance significantly in a short time. A training program for basic and advanced laparoscopic skills should be incorporated into the syllabus of urologists-in training and available to those who wish to gain experience in laparoscopic surgery. PMID- 12771715 TI - Comparison of quality of life following laparoscopic and open prostatectomy for prostate cancer. AB - PURPOSE: We compare the quality of life after laparoscopic prostatectomy to that after standard radical prostatectomy. MATERIAL AND METHODS: The quality of life of 52 and 54 patients who underwent laparoscopic and open radical prostatectomy, respectively, was analyzed using the European Organization for the Research and Treatment of Cancer Prostate Cancer quality of life questionnaire for general health related quality of life, International Index of Erectile Function 5 for screening erectile dysfunction and International Continence Society MaleSF questionnaire to evaluate urinary status. These questionnaires were given to patients before and 6 months after surgery. RESULTS: The general health related quality of life survey revealed no significant differences in health before and after laparoscopic and open prostatectomy. However, sexual quality of life was markedly lower after surgery (p <0.01). In addition, the International Index of Erectile Function score was markedly abrogated by surgery (p <0.05) and quality of life due to urinary incontinence was significantly disturbed by surgery (p <0.05). In contrast, quality of life due to voiding dysfunction was impaired before surgery and significantly improved by surgery (p <0.05). Patients were also asked if they would choose the same treatment if suffering from the same disease, with more patients treated laparoscopically choosing the same treatment than those treated with open surgery (p <0.05). CONCLUSIONS: While general health related quality of life was not impaired, sexual quality of life was diminished by surgery. Patients were generally satisfied with postoperative urinary status. Although patients who underwent laparoscopic prostatectomy expressed a more favorable attitude toward surgery, there was no significant difference in quality of life at 6 months after surgery between the 2 groups. PMID- 12771716 TI - Positive surgical margins in laparoscopic radical prostatectomy: the impact of apical dissection, bladder neck remodeling and nerve preservation. AB - PURPOSE: We assess the incidence and location of positive surgical margins in a consecutive series of patients who underwent laparoscopic radical prostatectomy. MATERIALS AND METHODS: Between May 1998 and September 2001, 235 men underwent laparoscopic radical prostatectomy at our institution. During this period modifications were made in the technique. We stopped preserving the puboprostatic ligaments during 1999 and stopped preserving the bladder neck during 2000. We compared the incidence of apical and bladder neck margins before and after these changes in surgical technique. Nerve sparing status was documented by the surgeon at the end of the operation and its effect on the incidence of lateral positive margins was evaluated. To concentrate on the surgically induced margins this study focused on pT2 cases. RESULTS: There were 169 pT2 cases and 32 (18.9%) positive margin locations. A constant decrease was noted in the overall incidence of positive margins but, while apical margins were the most abundant in 1998, posterolateral margins were most abundant in 2001. Avoiding bladder neck preservation eliminated positive bladder neck margins in 2001. The overall incidence of positive lateral surgical margins in pT2 cases treated with a nerve sparing procedure was 8.4%. CONCLUSIONS: The experience gained in this procedure led to a decrease in the incidence of positive margins. Wide resection of the bladder neck and cutting the puboprostatic ligaments decreased bladder neck and apical positive margins. Nerve preservation did not increase the incidence of positive margins. These technical modifications may improve the outcome of laparoscopic radical prostatectomy. PMID- 12771717 TI - Laparoscopic radical nephrectomy: incorporating advantages of hand assisted and standard laparoscopy. AB - PURPOSE: We present an approach to laparoscopic radical nephrectomy and intact specimen extraction, which incorporates hand assisted and standard laparoscopic techniques. MATERIALS AND METHODS: A refined approach to laparoscopic radical nephrectomy is described and our experience is reviewed. A low, muscle splitting Gibson incision is made just lateral to the rectus muscle and the hand port is inserted. A trocar is placed through the hand port and pneumoperitoneum is established. With the laparoscope in the hand port trocar 2 additional trocars are placed under direct vision. The laparoscope is then repositioned through the middle trocar and standard laparoscopic instruments are used through the other 2 trocars including the one in the hand port. If at any time during the procedure the surgeon believes the hand would be useful or needed, the trocar is removed from the hand port and the hand is inserted. RESULTS: This approach has been applied to 7 patients. Mean estimated blood loss was 200 cc (range 50 to 300) and mean operative time was 276.7 minutes (range 247 to 360). Mean specimen weight was 767 gm. (range 538 to 1,170). Pathologically 6 specimens were renal cell carcinoma (grades 2 to 4) and 1 was oncocytoma. Mean length of hospital stay was 3.71 days (range 2 to 7). There were no major complications. CONCLUSIONS: We believe that this approach enables the surgeon to incorporate the advantages of the hand assisted and standard laparoscopic approaches. PMID- 12771718 TI - Laparoscopy and urologic oncology--I now pronounce you man and wife. PMID- 12771719 TI - Mayo Clinic Scottsdale experience with laparoscopic nephron sparing surgery for renal tumors. AB - PURPOSE: Nephron sparing surgery is an accepted treatment for small renal masses, of which many have been detected incidentally due to the widespread use of advanced imaging techniques. We report our experience with laparoscopic nephron sparing surgery. MATERIALS AND METHODS: From May 2000 to May 2002 a total of 20 laparoscopic partial nephrectomies were performed in 19 patients. The kidney was mobilized to allow adequate dissection, hemostasis and inspection of the kidney. Cautery, a harmonic scalpel and a TissueLink (TissueLink Medical, Inc., Dover, New Hampshire) device were variably used for dissection and hemostasis. Further hemostasis was then achieved using an argon beam laser with Fibrillar (Fibrillar Ethicon, Somerville, New Jersey), fibrin glue or the TissueLink device. Intact removal and biopsy of the lesion base were done to assess margin status. RESULTS: Mean patient age was 66 years (range 41 to 80). Mean tumor size was 2.1 cm. (range 1 to 7) and average operative time was 130 minutes (range 60 to 210). Mean hospital stay was 2.2 days. Mean estimated blood loss was 120 ml. (range 20 to 400) and no blood transfusions or conversions to an open procedure were required. Complications included intraoperative fragmentation of a tumor in 1 case, postoperative dyspnea, postoperative bleeding and pneumonia in 1. CONCLUSIONS: Laparoscopic partial nephrectomy for small renal tumors was performed safely and effectively. Technique depended on the size and location of the mass. Long-term followup is required to compare cancer control with that of open nephron sparing surgery. PMID- 12771720 TI - Laparoscopic radical prostatectomy: decreasing the learning curve using a mentor initiated approach. AB - PURPOSE: Laparoscopic radical prostatectomy is being evaluated at several centers in the United States as a treatment option for localized prostate cancer. It is a technically difficult operation to perform with a steep learning curve. It has been stated that 50 procedures are necessary to satisfy the learning curve. To expedite performance and evaluation of laparoscopic radical prostatectomy a surgeon (mentor) who had performed 200 cases was invited to instruct a fellowship trained laparoscopist (trainee). MATERIALS AND METHODS: From March 2001 through September 2001 we performed 30 laparoscopic radical prostatectomies. The mentor performed the first 12 procedures with the trainee acting as assistant (group 1). The subsequent 18 procedures were performed by the trainee with the mentor acting as assistant (group 2). A final set of 20 procedures was performed by the trainee alone using 1 of 3 urological residents as the assistant (group 3). The transperitoneal approach was used and all suturing was intracorporeal. Preoperative data included prostate specific antigen, clinical stage, Gleason grade and median patient age. Intraoperative data included operative time, the blood loss/transfusion rate and intraoperative complications. Postoperative data included pathological stage, prostate specific antigen, the positive margin rate, catheter dwell time and hospital stay. When applicable, statistical significance was determined using the standard paired t test. RESULTS: There was no statistical difference in median operative time in groups 1 and 2 (248 and 258 minutes, respectively, p = 0.15). Similarly there was no difference in groups 2 (trainee and mentor assistant) and 3 (trainee alone) (p = 0.26). There was a difference in operative time in groups 1 and 3 (p = 0.04). Mean estimated blood loss was comparable in groups 1 to 3 and not statistically different (150, 250 and 250 cc, respectively, p = 0.15). Mean organ weight was also comparable (64, 59 and 55 gm., respectively). Hospital stay was 3 days in all groups. Catheter time decreased as confidence was gained with the procedure (range 6 to 33 days). Final pathological stage was compared among the 3 groups. There was an overall increase in positive margins in groups 1 to 3 (16%, 22% and 30%, respectively, p not significant). However, the positive margin rate for stage pT2 disease was similar at 15.5% for groups 1 and 2, and 14% for group 3. CONCLUSIONS: Laparoscopic radical prostatectomy is a technically challenging operation that is in the early stages of evolution and evaluation. We present an intensive, mentor initiated approach to decrease the learning curve and maintain outcomes. PMID- 12771721 TI - Endoscopic extraperitoneal radical prostatectomy: initial experience after 70 procedures. AB - PURPOSE: After our initial experience with 70 transperitoneal laparoscopic radical prostatectomies we developed a totally extraperitoneal retropubic approach to radical prostatectomy using laparoscopic instruments. We report our initial experience with 70 endoscopic extraperitoneal radical prostatectomy procedures. MATERIALS AND METHODS: A total of 70 patients underwent endoscopic extraperitoneal radical prostatectomy. Mean patient age was 63.4 years (range 49 to 76). Mean preoperative prostate specific antigen was 12.48 ng./ml. (range 1.4 to 50.7). There were no specific selection criteria for the procedure. The steps of the procedure are preparation of the preperitoneal space with the help of a balloon trocar, trocar placement (a 3 x 5 and a 2 x 12 mm. port), pelvic lymph node dissection, exposure of the prostate and the bladder neck, incision of the endopelvic fascia, ligation of Santorini's plexus, bladder neck dissection, mobilization of the seminal vesicles, incision of Denonvilliers' fascia, sectioning of the prostatic pedicles with or without preservation of the neurovascular bundles, dissection of Santorini's plexus and apex, urethrovesical anastomosis with 7 to 9 interrupted sutures and removal of the specimen via an extraction bag. During the 70 endoscopic prostatectomies 11 hernia defects were treated in 9 patients concomitantly. RESULTS: There was no conversions and no re interventions. Mean operative time was 155 minutes (range 90 to 260). One patient required transfusion with 2 units of blood cells. Pathological stage was pT2a in 19 patients, pT2b in 14, pT3a in 25, pT3b in 9 and pT4 in 3. Positive surgical margins were found in 2 of the 33 patients (6.1%) with pT2 tumors and in 13 of the 37 (35.1%) with pT3 and pT4 tumors. Postoperatively edema and hematoma of the penis in 10 cases was treated conservatively. Furthermore, 4 patients had asymptomatic lymphoceles, 1 required lymphocele drainage and 2 had partial obturator nerve paralysis, which resolved spontaneously. In 1 patient deep venous thrombosis developed. CONCLUSIONS: The preliminary results of this series are promising. Operative and perioperative morbidity was low. Functional results and oncological control were similar to the results of laparoscopic radical prostatectomy. The data demonstrate that endoscopic extraperitoneal radical prostatectomy can be performed with efficacy and results equal to those of laparoscopic radical prostatectomy, while providing the benefits of a totally extraperitoneal approach. Therefore, totally endoscopic extraperitoneal radical prostatectomy represents a technical improvement of laparoscopic technique because it completely obviates intra-abdominal complications and combines the advantages of minimally invasive laparoscopy and the retropubic open approach. PMID- 12771722 TI - Oncological safety of laparoscopic surgery for urological malignancy: experience with more than 1,000 operations. AB - PURPOSE: Although laparoscopy is being increasingly used to treat urological malignancies, there is still concern regarding the induction of local recurrence and port site metastasis. To our knowledge no major clinical study with long-term followup has been presented in the field of urological laparoscopy. We assessed the oncological safety of laparoscopy with emphasis on incidence of local recurrence and port site metastasis, analyzing the risk factors for such events based on a 10-year experience. MATERIALS AND METHODS: From June 1992 to May 2002 we performed 1,098 laparoscopic procedures for urological malignancies, including 450 radical prostatectomies, 478 pelvic and 80 retroperitoneal lymph node dissections, 45 radical nephrectomies, 22 radical nephroureterectomies, 12 partial nephrectomies and 11 adrenalectomies. In 418 cases of laparoscopic radical prostatectomy pelvic lymphadenectomy was done simultaneously. Of the procedures 917 were performed transperitoneally, including 181 via retroperitoneal or extraperitoneal access. A total of 567 procedures were performed in case of histologically proven cancer, whereas 531 represented only staging operations. RESULTS: Median followup was 58 months (range 4 to 127). Eight local recurrences were observed (0.73% overall, 1.41% of histologically proven cases). There were recurrences after nephroureterectomy for transitional cell carcinoma of the ureter in 1 patient, after radical nephrectomy for renal cell carcinoma in 1, growing teratoma after retroperitoneal lymph node dissection in 2, local recurrence of prostate cancer in 3 and after removal of an adrenal metastasis of melanoma in 1. Two port site metastases (0.18% overall, 0.35% of histologically proved cases) occurred, including metastasis of small cell lung carcinoma after adrenalectomy and a residual mass following 2 cycles of chemotherapy after retroperitoneal lymph node dissection. CONCLUSIONS: According to our experience the incidence of local recurrence and the risk of port site metastases is low and seems to be mainly related to the aggressiveness of the tumor and immunosuppression status of the patient, respectively rather than to technical aspects of the laparoscopic approach. PMID- 12771723 TI - Renal cell carcinoma with retroperitoneal lymph nodes: role of lymph node dissection. AB - PURPOSE: We better defined the benefits and morbidity of lymph node dissection in patients with localized renal cell carcinoma using the experience of patients treated at our institution. MATERIALS AND METHODS: A retrospective cohort study was performed with outcome assessment based on the chart review of demographic, clinical and pathological data in 1,087 patients with renal cell carcinoma treated at our institution. Patients with renal cell carcinoma who did not undergo nephrectomy as part of cancer treatment, those with bilateral disease and those for whom nodal status was unknown were not included in this study. A total of 900 patients meeting these criteria who underwent nephrectomy for unilateral renal cell carcinoma at our medical center form the principal study population. RESULTS: Positive lymph nodes were associated with larger, higher grade, locally advanced primary tumors that were more commonly associated with sarcomatoid features. Positive nodes were 3 to 4 times more common in patients with metastatic disease and the majority of these patients could be identified preoperatively. The survival of patients with regional lymph node involvement only was identical to that of patients with distant metastatic disease only. Patients with regional nodes and distant metastases had significantly inferior survival to those with either condition alone. In node negative cases lymph node dissection can be performed with no additional morbidity but it confers no survival advantage. In node positive cases lymph node dissection can also be performed safely but it is associated with improved survival and a trend toward an improved response to immunotherapy. CONCLUSIONS: Regional lymph node dissection is unnecessary in patients with clinically negative lymph nodes since it offers extremely limited staging information and no benefit in terms of decreasing disease recurrence or improving survival. In patients with positive lymph nodes lymph node dissection is associated with improved survival when it is performed in carefully selected patients undergoing cytoreductive nephrectomy and postoperative immunotherapy. When lymph nodes are present, they should be resected when technically feasible. PMID- 12771725 TI - Molecular detection of von Hippel-Lindau gene mutations in urine and lymph node samples in patients with renal cell carcinoma: potential biomarkers for early diagnosis and postoperative metastatic status. AB - PURPOSE: Organ confined renal cell carcinoma can be cured in the majority of patients, whereas more extensive lesions have a poor prognosis. Therefore, the development of a useful biomarker for early diagnosis as well as postoperative metastatic status would contribute to the appropriate therapy for renal cell carcinoma. To diagnose renal cell carcinoma preoperatively we developed a novel urinary test and detected occult lymph node micrometastasis using a molecular approach. MATERIALS AND METHODS: Urine samples were obtained preoperatively from 27 patients with renal cell carcinoma and von Hippel-Lindau (VHL) gene mutations in the tumors, and were analyzed for VHL gene mutations using a nested single strand conformational polymorphism analysis. Lymph nodes without evidence of histological metastasis were obtained from 15 patients with renal cell carcinoma and VHL gene mutations, and analyzed for VHL gene mutations using mutation specific nested reverse transcription polymerase chain reaction method. RESULTS: In urine samples 5 of 27 VHL gene mutations (18.5%) were found and each mutation pattern was the same as that detected in each renal cell carcinoma. One lymph node micrometastasis was found. CONCLUSIONS: These data indicate the presence of detectable levels of tumor derived DNA in the urine of patients with renal cell carcinoma and suggest that nested single strand conformational polymorphism analysis of VHL gene of urine samples provides a possible tool for the early detection of renal cell carcinoma. Furthermore, mutation specific nested reverse transcription polymerase chain reaction is useful to detect occult lymph node micrometastasis and may predict patients at risk for local recurrence. These 2 combined approaches using VHL gene mutations may contribute to the total therapy for and prognosis of renal cell carcinoma. PMID- 12771724 TI - Heterozygosity or homozygosity for 2 HLA class II haplotypes predict favorable outcomes for renal cell carcinoma treated with cytokine therapy. AB - PURPOSE: Durable responses to cytokine therapy occur in a small subset of patients with renal cell carcinoma. We determined if a common HLA genotype existed among these patients which might be associated with response and survival. MATERIALS AND METHODS: The study population consisted of 80 patients with metastatic renal cell carcinoma who had received cytokine therapy. DNA obtained from these patients was used for high resolution typing of HLA A, B, C, DRB1, DQA1 and DQB1 alleles. RESULTS: The class II alleles from patients with prolonged disease-free survival were predominantly composed of haplotype DRB1*0301/DQA1*0501/DQB1*0201 and DRB1*1501/DQA1*0102/DQB1*0602. The frequency of heterozygosity or homozygosity for these alleles was significantly greater in the good outcome group of patients than in those whose disease progressed during therapy. Heterozygosity or homozygosity at these loci was also associated with significant prolongation of survival. CONCLUSIONS: We conclude that heterozygosity or homozygosity for the class II haplotypes DRB1*0301/DQA1*0501/DQB1*0201 and DRB1*1501/DQA1*0102/DQB1*0602 is associated with durable response and survival in patients with metastatic renal cell carcinoma treated with cytokine therapy. PMID- 12771726 TI - Immunocytochemical analysis of prostate stem cell antigen as adjunct marker for detection of urothelial transitional cell carcinoma in voided urine specimens. AB - PURPOSE: Prostate stem cell antigen (PSCA), a homologue of the Ly-6/Thy-1 family of cell surface antigens, has been shown to be increased in a majority of human transitional cell carcinomas. We tested the possibility of using PSCA as an adjunct marker for urine cytology. MATERIALS AND METHODS: Immunocytochemical analysis was performed on 44 archived voided urine samples obtained from 3 groups of patients based on initial voided urine cytological results and subsequent followup biopsy findings. Group 1 (14 of 44 patients) had positive findings on cytology and histology, group 2 (16 of 44) had negative cytology but positive histology, and group 3 (14 of 44) had negative findings on cytology and histology. Cytological slides prepared from 10 fresh voided urine samples were also analyzed. Papanicoloau stained archived urine slides were de-stained and re stained immunocytochemically with a monoclonal antibody against PSCA. Immunofluorescence followed by laser scanning cytometer analysis was also performed on archived slides from 2 representative cases. RESULTS: Sensitivity and specificity were 46.7% and 100% for cytology alone, respectively, and 80% and 85.7% for PSCA alone, respectively. PSCA immunostaining was positive in 92.8% group 1, 68.8% group 2 and only 14.3% group 3 samples. The difference in positive PSCA findings in groups 2 and 3 were statistically significant at p <0.01 by chi square test. Whereas some superficial umbrella cells showed slight staining by immunocytochemistry, it was feasible to distinguish the expression levels between tumor and normal superficial umbrella cells quantitatively using immunofluorescence coupled with laser scan cytometry analysis. CONCLUSIONS: Immunocytochemical analysis of PSCA on archived voided urine samples may provide a simple and quantitative adjunct marker for cytological diagnosis of urothelial carcinoma. PMID- 12771727 TI - Multitarget fluorescence in situ hybridization assay detects transitional cell carcinoma in the majority of patients with bladder cancer and atypical or negative urine cytology. AB - PURPOSE: The multitarget fluorescence in situ hybridization (FISH) probe set UroVysion (Vysis, Downers Grove, Illinois), containing probes to chromosomes 3, 7 and 17, and to the 9p21 band, has been recently shown to have high sensitivity and specificity for detecting transitional cell carcinoma. In this study we retrospectively tested 120 urine samples from patients with atypical, suspicious and negative cytology for whom concurrent and followup bladder biopsy data were available. We evaluated the ability of FISH to identify malignant cells in cytologically equivocal or negative cases. MATERIALS AND METHODS: Archived slides from 120 voided (47) or instrumented (73) urine cytology specimens from patients with concurrent bladder biopsy and a minimum of 12 months of biopsy followup were subjected to hybridization with UroVysion. The cohort included patients with biopsy proven transitional cell carcinoma, which was grades 1 to 3 in 23, 35 and 24, respectively, and stages pTis in 3, pTa in 64, pT1 in 6, pT2 in 6 and pT4 in 3, while it showed negative histology in 38. Cytology findings were suspicious, atypical and negative for transitional cell carcinoma in 31, 49 and 40 cases, respectively. A positive FISH result was defined as 5 transitional cells or greater with a gain of 2 or more of chromosomes 3, 7 or 17, 12 cells or greater with 9p21 deletion, or 10% or greater of cells with isolated trisomy of 1 of chromosomes 3, 7 and 17. RESULTS: All except 12 of the 82 biopsy proven transitional cell carcinoma cases (11 pTa and 1 pT1 tumors) were positive by FISH (85% sensitivity). Sensitivity in patients with suspicious, atypical and negative cytology was 100%, 89% and 60%, respectively. Nine patients with atypical cytology had positive FISH in the setting of a negative concurrent bladder biopsy. However, 8 of these 9 patients (89%) had biopsy proven transitional cell carcinoma within 12 months following the date when the sample tested by FISH was obtained. The last of these patients with false-positive results had previously documented pTis disease, which was also present in the next bladder biopsy 15 months following the positive FISH result. The remaining 29 specimens from patients with negative biopsy and a negative 12-month followup tested negative by FISH (97% overall specificity). CONCLUSIONS: The UroVysion FISH assay provides high sensitivity and specificity to detect transitional cell carcinoma in cytologically equivocal and negative urine samples. These results emphasize the important role of this assay in the management of bladder cancer. PMID- 12771728 TI - Grade progression and regression in recurrent urothelial cancer. AB - PURPOSE: Recurrent urothelial cancers are reported to have characteristics similar to those of the primary tumor, with 10% to 25% of low grade tumors recurring as high grade disease. We determined how often grade progression and regression occur and whether abnormalities in p53 protein expression in original tumors are preserved in recurrences. MATERIALS AND METHODS: Two groups of patients treated for recurrent stages Ta/T1 urothelial bladder cancers with at least 1 tumor-free examination between the index and recurrent tumors were reviewed. Group 1 included 115 patients in whom the first available tumor was compared with the last recurrence and group 2 included 42 in whom the initial tumor was compared with the first recurrence. Immunohistochemical analysis of p53 expression was performed on a subset of 34 tumor pairs. RESULTS: In group 1, 33 grade 3 tumors (45%) recurred as grade 1 or 2 tumors, while 9 of 82 grades 1 and 2 tumors (11%) recurred as grade 3 tumors. Five of 7 group 2 grade 3 tumors (71%) recurred as grade 1 or 2 disease, while 1 of 35 grades 1 and 2 tumors (3%) recurred as grade 3 disease. In the 34 pairs studied immunohistochemically 6 of 14 grade 3 tumors recurred at lower grades. Nuclear p53 over expression occurred in 21 index tumors (12 of 14 grade 3, 8 of 17 grade 2 and 1 of 3 grade 1) and in 9 recurrences (6 of 10 grade 3, 2 of 17 grade 2 and 1 of 7 grade 1). Only 7 of 21 p53 positive and 2 of 12 p53 negative index tumors were p53 positive on recurrence. CONCLUSIONS: While progression from low to high grade occurred in less than 15% of patients, grade regression was observed in almost 50%. The loss of p53 positivity in regressing tumors indicates that these recurrences are molecularly distinct from the corresponding initial tumor. PMID- 12771729 TI - Intravesical bacillus Calmette-Guerin therapy for stage T1 grade 3 transitional cell carcinoma of the bladder: recurrence, progression and survival in a study of 57 patients. AB - PURPOSE: Stage T1 grade 3 transitional cell carcinoma of the bladder is associated with a high risk of tumor recurrence and progression. We report our experience with stage T1 grade 3 bladder tumors treated with bacillus Calmette Guerin (BCG) therapy in the last 10 years. MATERIALS AND METHODS: We analyzed the outcome in 57 consecutive patients treated with intravesical BCG for stage T1 grade 3 bladder cancer between 1991 and 2001. After initial transurethral resection all patients received a 6-week course of BCG therapy consisting of 1 instillation weekly. All patients underwent systematic biopsies at the end of the first BCG course. Patients with negative biopsies received maintenance BCG therapy, consisting of intravesical instillations each week for 3 weeks given 3, 6, 12, 18, 24, 30 and 36 months after the first course. Patients with residual tumor received a second course of 6 weekly instillations. Time to tumor recurrence and progression, and the rate of patient survival were retrospectively analyzed. RESULTS: Median followup was 53 months (range 9 to 110). Minimum followup was 2 years in 36 cases (63.2%) and 5 years in 28 (49.1%). After the first BCG course 50 patients (87.7%) had no residual disease, while 7 (12.3%) had residual tumor. The recurrence and progression rates were 42.1% and 22.8%, respectively. The rate of delayed cystectomy was 14%. The rate of disease specific survival was 87.7%. CONCLUSIONS: Our study confirms that BCG therapy is effective conservative treatment for patients with stage T1 grade 3 bladder tumors. PMID- 12771730 TI - Is there a therapeutic role for post-chemotherapy retroperitoneal lymph node dissection in metastatic transitional cell carcinoma of the bladder? AB - PURPOSE: We identified a subset of patients with bladder cancer (transitional cell carcinoma) and regional nodal metastasis to the retroperitoneal lymph nodes without detectable systemic dissemination. While the majority of these patients respond initially to chemotherapy, most have disease relapse at the same site within a year. We report the results of a phase II study exploring the potential benefit of retroperitoneal lymph node dissection in patients with transitional cell carcinoma of the bladder in whom disease has shown a significant response to chemotherapy. MATERIALS AND METHODS: A total of 11 patients with biopsy proven metastatic transitional cell carcinoma in the retroperitoneal lymph nodes and no evidence of visceral metastatic disease in whom disease showed a significant response to chemotherapy underwent complete bilateral retroperitoneal lymph node dissection. The end point of study was disease specific survival, calculated from the time of retroperitoneal lymph node dissection to death from transitional cell carcinoma of the bladder. RESULTS: Four patients underwent delayed retroperitoneal lymph node dissection. Seven patients underwent concurrent cystectomy, and pelvic and retroperitoneal lymph node dissection. There was no perioperative mortality. Nine patients had evidence of residual disease in the retroperitoneal nodes. Seven patients have recurrence outside of the original surgical field with a median time to recurrence of 7 months and 6 died at a median time to death of 8 months (range 5 to 14). One patient with retrocrural recurrence attained a complete response to salvage chemotherapy and remained disease-free 57 months after retroperitoneal lymph node dissection. For all 11 patients median disease specific and recurrence-free survival rates were 14 and 7 months, respectively. Four-year disease specific and recurrence-free survival rates were 36% and 27%, respectively. We stratified the patients based on the number of involved lymph nodes at retroperitoneal lymph node dissection and noted that viable tumor in no more than 2 lymph nodes correlated with greater disease specific and recurrence-free survival (p = 0.006 and 0.01, respectively). CONCLUSIONS: Retroperitoneal lymph node dissection can be safely performed for metastatic transitional cell carcinoma. Retroperitoneal lymph node dissection has curative potential, particularly in patients with viable tumor in no more than 2 lymph nodes after chemotherapy. PMID- 12771731 TI - Combined laser treatment for penile carcinoma: results after long-term followup. AB - PURPOSE: We evaluated local disease control, side effects and cause specific survival of penile carcinoma treated with laser therapy. MATERIALS AND METHODS: In a prospective study from 1986 to 2002 we included 67 men with a mean age of 60 years with newly diagnosed penile carcinoma. No patient was lost to followup. RESULTS: At a median followup of 42 months (range 12 to 186) 59 patients were alive and 8 had died of penile carcinoma (2) and concurrent disease (6). Of the 13 patients (19%) with local recurrence during the study period 10 underwent repeat laser treatment successfully. Side effects were few but 5 patients (7%) had postoperative bleeding. Overall cosmetic and functional results were highly satisfactory. CONCLUSIONS: The results of this study show that treating penile carcinoma with the combination of carbon dioxide and neodymium:YAG lasers can safely be done with highly satisfactory cosmetic results as well as good local tumor control. PMID- 12771732 TI - Histology and clinical outcomes in patients with bilateral testicular germ cell tumors: the Memorial Sloan Kettering Cancer Center experience 1950 to 2001. AB - PURPOSE: The optimal management of bilateral testicular tumors continues to evolve. We examined the incidence, characteristics, histology, treatment and clinical outcomes of patients with bilateral testicular cancer. MATERIALS AND METHODS: Between 1950 and 2001, 3,984 patients with testicular cancer were treated at our center. A total of 58 patients with bilateral testicular germ cell tumors were identified. The clinical records of these 58 patients were reviewed for age, histology of the 2 tumors, stage at presentation of the first and second tumor, interval between tumors, treatment and clinical outcome. Median followup was 60 months. RESULTS: Ten of the 58 patients (17%) had synchronous tumors, while the other 48 (83%) had metachronous tumors with a median interval between tumors of 50.5 months. Overall seminoma was the most common histology of the synchronous and metachronous tumors. Most patients in the synchronous and metachronous tumor groups presented with low stage disease. Of the 58 patients 52 (89%) had no evidence of disease and 6 (11%) were dead of disease at the last followup. Treatment of the second tumor appeared to be influenced by therapy for the first tumor in 16.7% of cases. CONCLUSIONS: Patients with a history of testicular germ cell tumor require careful long-term monitoring of the contralateral testicle due to the risk of bilateral disease and potentially long latent period between the first and second tumors. Overall the clinical outcome is good in these patients when they are treated appropriately for histology and stage. In patients with metachronous tumors treatment of the contralateral tumor is rarely altered by prior treatment of the initial tumor. PMID- 12771733 TI - Is post-chemotherapy resection of seminomatous elements associated with higher acute morbidity? AB - PURPOSE: A seminomatous element in patients undergoing retroperitoneal lymph node dissection for testicular cancer is associated with a desmoplastic reaction that renders retroperitoneal surgery more challenging. We examined the impact of seminomatous elements on the rate of complications and the need for additional intraoperative procedures in patients undergoing post-chemotherapy retroperitoneal lymph node dissection. MATERIALS AND METHODS: The testis cancer data base at our institution was retrospectively reviewed and 1,366 patients were identified who underwent post-chemotherapy retroperitoneal lymph node dissection between 1973 and 2001. In 97 patients there was an element of seminoma in the dissection specimen and/or pure seminoma in the testicular primary specimen (seminoma group). The remaining 1,269 patients underwent post-chemotherapy retroperitoneal lymph node dissection for nonseminomatous testicular tumors. The rates of intraoperative complications and additional procedures as well as postoperative complications were analyzed. RESULTS: Of the 97 patients in the seminoma group 37 (38.1%) required a total of 47 additional intraoperative procedures, including 25 nephrectomies, 9 inferior vena caval resections, 5 arterial grafts, 5 bowel resections and 3 hepatic resections/biopsies, compared with 340 of the 1,269 patients (26.8%) in the group without seminomatous elements (p = 0.02). Postoperatively complications occurred in 24 of 97 patients (24.7%) in the seminoma group versus 257 of 1,269 (20.3%) in the group without seminomatous elements (p = 0.29). One of the 97 patients in the seminoma group died secondary to postoperative complications. CONCLUSIONS: A seminomatous element in patients undergoing post-chemotherapy retroperitoneal lymph node dissection is associated with a higher rate of additional intraoperative procedures and postoperative complications than in patients without seminomatous elements. However, resection is still possible with acceptable morbidity when indicated in appropriately selected patients. PMID- 12771734 TI - Improved clinical staging system combining biopsy laterality and TNM stage for men with T1c and T2 prostate cancer: results from the SEARCH database. AB - PURPOSE: A number of studies have failed to show significant differences in outcome following radical prostatectomy between men with palpable, clinically localized prostate cancer (cT2) and those whose tumors are not palpable (cT1c). We determined whether we could improve the prognostic value of the TNM staging system in men with cT1c and cT2 cancers by including information on whether prostate needle biopsy was unilaterally or bilaterally positive. MATERIALS AND METHODS: A retrospective survey of 992 patients from the SEARCH (Shared Equal Access Regional Cancer Hospital) Database treated with radical prostatectomy at 4 equal access medical centers between 1988 and 2002 was done. TNM 1992 clinical stage was T1c in 421 patients, T2a in 287, T2b in 202 and T2c in 82. Multivariate analysis was used to examine whether biopsy laterality and clinical stage were significant predictors of surgical margin status, nonorgan confined disease, seminal vesicle invasion, and time to prostate specific antigen (PSA) recurrence following radical prostatectomy. RESULTS: Patients with clinical stages T2b and T2c cancers had similar rates of PSA recurrence, which were significantly higher than in patients with T1c and T2a disease, who also had similar rates of PSA recurrence. Bilateral positive biopsy further stratified patients with T1c and T2a disease (p = 0.01) but not those with T2b and T2c cancers (p = 0.207). Grouping these 1992 clinical stages with biopsy laterality resulted in a new clinical staging system, which was a significant predictor of PSA recurrence following radical prostatectomy (p <0.001). On multivariate analysis whether TNM clinical stage was evaluated as a categorical or continuous variable only PSA, biopsy Gleason score and the new clinical staging system (1992 TNM stage groupings combined with biopsy laterality) were significant independent predictors of time to biochemical recurrence following radical prostatectomy. CONCLUSIONS: Combining low (T1c and T2a) and high (T2b and T2c) risk 1992 clinical stages with biopsy laterality (unilateral versus bilateral positive) resulted in a new clinical staging system that was a stronger predictor of PSA recurrence following radical prostatectomy than the 1992 or 1997 TNM clinical staging system. If confirmed at other centers and in men who undergo with other treatment modalities, consideration should be given to revising the current TNM staging system to reflect these findings. PMID- 12771735 TI - Percent of prostate needle biopsy cores with cancer is significant independent predictor of prostate specific antigen recurrence following radical prostatectomy: results from SEARCH database. AB - PURPOSE: Recent studies have suggested that the percent of positive cores in the prostate needle biopsy is a significant predictor of outcome among men undergoing radical prostatectomy or radiation therapy for prostate cancer. We evaluate whether either percent of cores with cancer or percent of cores positive from the most and least involved side of the prostate needle biopsy was associated with a worse outcome among men treated with radical prostatectomy. MATERIALS AND METHODS: A retrospective survey of 1,094 patients from the SEARCH Database treated with radical prostatectomy at 4 different equal access medical centers in California between 1988 and 2002 was undertaken. We used multivariate analysis to examine whether total percent of prostate needle biopsy cores with cancer, percent of cores positive from each side of the prostate and other clinical variables were significant predictors of adverse pathology and time to prostate specific antigen (PSA) recurrence following radical prostatectomy. RESULTS: On multivariate analysis serum PSA and percent of positive cores were significant predictors of positive surgical margins, nonorgan confined disease and seminal vesicle invasion. Percent of positive cores (p <0.001), serum PSA (p = 0.008) and biopsy Gleason score (p = 0.014) were significant independent predictors of time to biochemical recurrence. On a separate multivariate analysis that included the variables of total percent of positive cores, percent of positive cores from the most involved side of the biopsy, percent of positive cores from the least involved side of the biopsy and whether the biopsy was positive unilaterally or bilaterally, only the percent of positive cores from the most involved side of the biopsy was a significant independent predictor of PSA failure following radical prostatectomy. Percent of positive cores was used to separate patients into a low risk (less than 34%), intermediate risk (34% to 50%) and high risk (greater than 50%) groups, which provided significant preoperative risk stratification for PSA recurrence following radical prostatectomy (p <0.001). Percent of positive cores cut points were able to further risk stratify men who were at low (p = 0.001) or intermediate (p = 0.036) but not high (p = 0.674) risk for biochemical failure based on serum PSA and biopsy Gleason score. CONCLUSIONS: Percent of positive cores in the prostate needle biopsy was a significant predictor of adverse pathology and biochemical failure following radical prostatectomy, and the cut points of less than 34%, 34% to 50% and greater than 50% can be used to risk stratify patients preoperatively. The finding that percent of positive cores from the most involved side of the biopsy was a stronger predictor of PSA failure than the total percent of cores involved suggests that multiple positive biopsies from a single side might be a better predictor of a larger total cancer volume and thus correlate with clinical outcome. PMID- 12771736 TI - Should a positive surgical margin following radical prostatectomy be pathological stage T2 or T3? Results from the SEARCH database. AB - PURPOSE: The finding of a positive surgical margin associated with extracapsular extension at radical prostatectomy is a poor prognostic factor. However, whether a positive surgical margin with no documented extracapsular extension portends a similarly poor prognosis is unclear. We examined the significance of the pathological features of positive surgical margin and extracapsular extension for predicting biochemical failure following radical prostatectomy. MATERIALS AND METHODS: We examined data on 1,621 men from the SEARCH Database of patients treated with radical prostatectomy without lymph node metastasis. Patients were separated into 5 groups based on the pathological findings of positive surgical margin, extracapsular extension, and/or seminal vesicle invasion. Preoperative clinical variables were compared across the groups and the groups were compared for time to biochemical recurrence using Cox proportional hazards analysis. RESULTS: Men with seminal vesicle invasion had the highest prostate specific antigen (PSA) recurrence rates, while men with a negative surgical margin and no extracapsular extension had the lowest PSA recurrence rates. There were no differences in PSA failure rates between men with a positive surgical margin and no extracapsular extension versus men with a negative surgical margin and extracapsular extension versus men with extracapsular extension and a positive surgical margin. In this subset of patients with a positive surgical margin and/or extracapsular extension but no seminal vesicle invasion only serum PSA was a significant independent predictor of biochemical recurrence. CONCLUSIONS: Men with a positive surgical margin but no extracapsular extension had PSA recurrence rates similar to those in men with extracapsular extension with or without positive margins. Men with extracapsular extension had similar biochemical recurrence rates whether the surgical margin was positive or negative. If confirmed at other institutions, consideration should be given to modifying the current TNM staging system to reflect these findings. PMID- 12771737 TI - The New York University nerve sparing algorithm decreases the rate of positive surgical margins following radical retropubic prostatectomy. AB - PURPOSE: We developed an algorithm that prospectively defines when to excise the neurovascular bundles during radical retropubic prostatectomy with the goal of maximizing the performance of nerve sparing procedures while minimizing positive surgical margins. MATERIALS AND METHODS: From January 1 to December 31, 2000 a single surgeon performed 272 radical retropubic prostatectomies and 263 were performed from January 1 to December 31, 2001. A single pathologist analyzed all specimens with positive margins. There were no prospectively defined criteria to guide decisions regarding excision of the neurovascular bundles in the 2000 study cohort. Gleason score, percent tumor volume and perineural invasion were independently analyzed in the biopsy specimens according to the site of origin (right versus left side) for the 2001 group only. The ipsilateral neurovascular bundle was excised for Gleason 6 or less tumors when there were 50% or greater tumor volume in the biopsy specimen and perineural invasion, for Gleason 7 tumors when there was 30% or greater tumor volume, or perineural invasion and for Gleason 8 to 10 tumors when there was 10% or greater tumor volume, or perineural invasion. RESULTS: There were no statistically significant differences between the 2000 and 2001 groups in regard to preoperative prostate specific antigen, clinical and pathological stage, biopsy Gleason score and percent tumor volume in the surgical specimen. There was a statistically significant decrease in the incidence of positive margins between the 2000 and 2001 groups (14% versus 8%, p = 0.027). The lower positive margin rate was not achieved because of a tendency to excise more neurovascular bundles since a significantly greater percent of neurovascular bundles was preserved in the 2001 group. The sensitivity, specificity, positive and negative predictive values, and accuracy of our algorithm were 18%, 93%, 28%, 89% and 84%, respectively. In sides of the prostate with extraprostatic extension ipsilateral wide excision of the neurovascular bundle was associated with positive margins in 33% of cases compared with 22% when the neurovascular bundle was preserved (p = 0.42). CONCLUSIONS: The New York University nerve sparing algorithm prospectively defines when to excise the neurovascular bundle based on Gleason score, perineural invasion and tumor volume in the biopsy specimen. Use of this algorithm decreases positive surgical margin rates, while significantly increasing the preservation of neurovascular bundles. PMID- 12771738 TI - Modifying the American Society for Therapeutic Radiology and Oncology definition of biochemical failure to minimize the influence of backdating in patients with prostate cancer treated with 3-dimensional conformal radiation therapy alone. AB - PURPOSE: Adoption of the American Society for Therapeutic Radiology and Oncology (ASTRO) consensus definition has been critical for evaluating and comparing outcome following treatment with radiation. However, since its almost universal adoption, several points have remained controversial, notably backdating the date of failure to the point midway between the posttreatment prostate specific antigen (PSA) nadir and the first increase. We evaluated the impact of backdating on no biochemical evidence of disease (bNED) control and suggest changes in the definition. MATERIALS AND METHODS: Between April 1, 1989 and November 30, 1998, 1,017 patients with nonmetastatic prostate cancer were treated with 3-dimensional conformal radiation therapy alone. bNED control was defined using the ASTRO consensus definition. bNED failure was calculated from the time midway between the posttreatment PSA nadir and the first of the 3 consecutive increases in PSA (date of failure A). Four alternate failure time points were chosen, including backdating to the date of the first increase in PSA after the nadir, the date between the first and second consecutive PSA increases, the date between the second and third consecutive PSA increases, and the date of the third increase in PSA after the nadir (dates of failure 1 to 4). Kaplan-Meier estimates were calculated for all definitions of failure as well as hazard functions with time. Subset analyses based on prognostic group and followup time were also performed. RESULTS: The 10-year Kaplan-Meier bNED control rates were 64%, 52%, 47%, 42% and 39% using dates of failure A and 1 to 4, respectively. These differences persisted when patients were stratified by prognostic group. These same differences in bNED control were observed for the long-term followup subset, in which 10-year bNED control rates were 48%, 47%, 44%, 41% and 39% using dates of failure A and 1 to 4, respectively. CONCLUSIONS: Adoption of the ASTRO consensus definition has been crucial for evaluating outcome in the radiation oncology community. However, the date of failure should be moved from the current point to one closer to the point at which failure is declared. Additional analysis with large numbers of patients from multiple institutions is necessary to determine the point. PMID- 12771739 TI - Hazard rates of disease progression after external beam radiotherapy for clinically localized carcinoma of the prostate. AB - PURPOSE: We established hazard rates for disease progression at different intervals after external beam radiotherapy alone in patients with clinically localized prostate cancer. We determined the likelihood of biochemical failure in those free of disease 5 years after radiotherapy and identified those at risk for early versus late biochemical failure. MATERIALS AND METHODS: We reviewed the records of 964 patients treated with full dose external beam radiotherapy alone for T1 to T4, NxM0 prostate cancer. Followup prostate specific antigen (PSA) was measured 3 months after the completion of radiotherapy and every 3 to 6 months thereafter. Yearly hazard rates for biochemical failure were calculated each followup year. RESULTS: Median followup of the whole study group was 48 months. Overall 5 and 10-year biochemical disease-free survival rates were 63.7% and 53.6%, respectively. Patients had a peak overall hazard rate 2 years after treatment. The hazard rate then decreased until year 6, when it increased slightly and remained elevated even at year 10. This late increase in the overall hazard rate was associated with late increases in the hazard rate in men with favorable prognostic factors, namely pretreatment PSA less than 10 ng./ml., Gleason score less than 5, clinical T stage T1 or T2, posttreatment PSA nadir less than 0.99 ng./ml. or radiation dose less than 68 Gy. In 13 of the 307 patients (3.9%) with biochemical failure the failure occurred more than 5 years after initial treatment. CONCLUSIONS: The peak risk of biochemical failure is 2 years after radiotherapy. Patients are at slight but persistent risk for biochemical failure more than 5 years after treatment. Hazard rate calculations beyond the median followup of 4 years can underestimate the true hazard. PMID- 12771741 TI - Early catheter removal after radical retropubic prostatectomy: long-term followup. AB - PURPOSE: We examine the complication and continence rates with early catheter removal (day 3 or 4) after radical retropubic prostatectomy. MATERIALS AND METHODS: A total of 365 patients with localized prostate cancer underwent radical retropubic prostatectomy at Indiana University Hospital with planned urethral catheter removal before discharge home. Low pressure cystograms were performed on postoperative day 3 or 4 to determine if catheter removal was possible. A subset of patients were analyzed using a validated prostate cancer specific questionnaire (University of California, Los Angeles Prostate Cancer Symptom Index) to determine quality of life outcomes. RESULTS: The catheter was removed on postoperative day 3 or 4 in 263 patients (72%). The reasons for leaving the catheter indwelling were significant leak on cystogram or excessive suprapubic drainage (21%), extensive bladder neck reconstruction (1%) and prolonged hospitalization because of an ileus or other complicating factor (6%). Thirteen patients (3.6%) were either unable to void after catheter removal or presented with retention (not associated with hematuria or clots) after hospital discharge, requiring reinsertion of the Foley catheter. A total of 41 patients (11%) had either an early or late complication (excluding incontinence). There were 3 complications (0.8%) that were considered major because they were potentially life threatening or required a return to the operating room. A pelvic abscess developed in 2 patients and a lymphocele in 1, which required percutaneous drainage. After at least 6 months (mean 20.9 months) 140 patients (89.2%) and 14 (8.9%) reported excellent and good continence, respectively. The patient questionnaire demonstrated bother scores to be minimal to no bother for 95% to 98% of patients at 6 and 12 months. CONCLUSIONS: This study confirms that it is safe to remove catheters in most patients 3 to 4 days after prostatectomy if a cystogram demonstrates no extravasation. Complication rates and continence rates with this approach compare favorably with series in which catheters are left indwelling for longer periods. PMID- 12771742 TI - The incidence and causes of erectile dysfunction after pelvic fractures associated with posterior urethral disruption. AB - PURPOSE: Erectile dysfunction is a common sequel of pelvic fractures, particularly those associated with posterior urethral injury when it can be neurogenic or arteriogenic due to damage to the cavernous nerves or branches of the pudendal arteries. We studied erectile function of patients with posterior urethral injuries due to pelvic fractures. MATERIALS AND METHODS: Patients referred for posterior urethral reconstruction and strictures due to pelvic fractures were evaluated before reconstruction. All patients underwent nocturnal penile tumescence testing, and if those results were abnormal, penile duplex ultrasound with intracavernous injection was performed. Patients with normal vascular function on duplex ultrasound were diagnosed with neurogenic erectile dysfunction. Those patients with abnormal arterial function on duplex ultrasound underwent arteriography to further define the extent and location of arterial damage. RESULTS: The study included 25 consecutive patients with posterior urethral strictures and a mean age of 28.6 years. Of the patients 18 (72%) had erectile dysfunction as demonstrated by nocturnal penile tumescence and all underwent penile duplex ultrasound. Ultrasound confirmed normal vascular response in 13 of the 18 patients and they were diagnosed with probable neurogenic erectile dysfunction. The remaining 5 patients (28%) with erectile dysfunction had an abnormal arterial response, and significant arterial pathology was confirmed by arteriography. CONCLUSIONS: Erectile dysfunction is common in patients with pelvic fractures associated with urethral injury. We believe that erectile function should be assessed and documented in such patients before attempting urethroplasty. In the majority of these patients erectile dysfunction is caused by disruption of the cavernous nerves with sparing of arterial inflow. PMID- 12771740 TI - Is there an optimal time for intracavernous prostaglandin E1 rehabilitation following nonnerve sparing radical prostatectomy? Results from a hemodynamic prospective study. AB - PURPOSE: Previous studies have shown that early intracavernous prostaglandin E1 injection may reduce significantly the incidence of veno-occlusive dysfunction before spontaneous erections recover after nerve sparing radical prostatectomy. We identify the more convenient postoperative timing for successful intracavernous injection rehabilitation in a series of patients who underwent nonnerve sparing radical prostatectomy. MATERIALS AND METHODS: A total of 73 patients with a normal preoperative International Index of Erectile Function score were randomly allocated to undergo dynamic color Doppler ultrasound study 20 mg. prostaglandin E1 at 1, 2 to 3, 4 to 6 and 7 to 12 months postoperatively, respectively. In all cases the peak systolic velocity, end diastolic velocity and resistance index were evaluated at 5, 10 and 20 minutes after injection. RESULTS: Of the patients 36 received the intracavernous injection within the first 3 months (group 1) and 37 received it at 4 to 12 months (group 2). A significantly higher proportion of group 1 patients had grade 3 erection compared with group 2. Peak systolic velocity less than 30 cm. per second in at least 1 cavernosal artery was recorded in 22.2% of group 1 patients and 51.3% of group 2 (p >0.05). CONCLUSIONS: Intracavernous injections after nonnerve sparing radical prostatectomy produce valid erectile responses in a significantly higher proportion of patients when started within month 3 after the operation. Injection given in postoperative month 1 gives the best response rate but with significant complications and poor patient compliance. Arteriogenic and venogenic factors seem to be involved with failure. PMID- 12771743 TI - The significance of measuring the time course of serum malondialdehyde concentration in patients with torsion of the testis. AB - PURPOSE: We determined the time course of malondialdehyde, a measure of free radical damage, in patients undergoing standard surgical treatment for testicular torsion. MATERIALS AND METHODS: Patients presenting with testicular torsion were studied prospectively. Blood samples were obtained after administering general anesthesia but before surgical incision, and 10 minutes, 30 minutes and 24 hours after detorsion. Orchiopexy was performed in patients with viable testes (group 1) and orchiectomy was performed in those with nonviable testes (group 2). Further blood samples were obtained 1 and 3 months after surgery. Similar blood samples were taken from controls, including patients younger than 40 years undergoing other operations involving manipulation of the testis, such as hydrocelectomy or orchiopexy (group 3). The level of malondialdehyde in each serum sample was determined by the thiobarbituric acid reaction. RESULTS: A total of 65 patients were studied, including 56 with testicular torsion and 9 controls (group 3). Of the 56 patients 11 (19.6%) with testicular torsion underwent ipsilateral orchiectomy and contralateral orchiopexy (group 2). The remaining 45 patients (80.4%) underwent bilateral orchiopexy (group 1). However serum malondialdehyde was estimated in only 34 of the 56 patients with torsion. Mean malondialdehyde at 0, 10 and 30 minutes, 24 hours, and 3 and 6 months was 3.3, 3.69, 3.69, 2.9, 2.65 and 2.39 nmol./ml. on the 24 group 1 patients, 3.53, 4.56, 3.87, 2.87, 2.82 and 2.64 nmol./ml. in the 10 group 2 patients, and 3.6, 3.08, 3.18, 2.95, 2.88 and 2.65 nmol./ml. in the 9 group 3 controls, respectively. The highest serum malondialdehyde was at 10 minutes after detorsion in groups 1 and 2. There was a statistically significant difference in malondialdehyde between groups 1 and 2 compared with group 3 at 10 minutes (p <0.04). Serum malondialdehyde returned to baseline at 24 hours in all patients. CONCLUSIONS: The results of this study indicate that testicular torsion and its treatment with detorsion is an example of ischemia-reperfusion injury, producing measurable changes in malondialdehyde in humans. Thus, serum malondialdehyde could be used to determine the extent of injury. PMID- 12771744 TI - Urethrorectal fistula: transanal, transsphincteric approach with locally based pedicle interposition flaps. AB - PURPOSE: We report preliminary experience with the transsphincteric, transanal surgical approach to correct acquired urethrorectal fistula. MATERIALS AND METHODS: Five men with an acquired urethrorectal fistula underwent surgical correction. Mean patient age was 56.6 years (range 37 to 72). The etiology was surgical (radical prostatectomy) in 3 cases, traumatic in 1 and idiopathic in 1. All 5 patients had urinary tract infection and pneumaturia, and 3 (60%) had watery stool. Conservative treatments had failed in all cases. The time from the diagnosis of urethrorectal fistula to surgery was 4 weeks to 4 years. RESULTSFive men underwent excision and closure of a urethrorectal fistula with diverting colostomy. In 4 men (80%) urinary continence subsequently returned with adequate sphincter tone, while in 1 (20%) with perineal trauma and active proctitis the fistula recurred 6 weeks after surgery. CONCLUSIONS: Urethrorectal fistulas are rare and surgically challenging. The transsphincteric, transanal surgical approach provides many advantages, including easy access and identification of the fistula tract, good surgical exposure, adequate resection to well vascularized tissue and access to several vascularized flaps for interposition between the repaired urinary and gastrointestinal tracts. PMID- 12771745 TI - Continent lower urinary tract reconstruction in the cervical spinal cord injured population. AB - PURPOSE: Patients with cervical spinal cord injury and upper extremity dysfunction are treated primarily with indwelling or condom catheters. We present our experience with a select group of patients with limited upper extremity function to determine long-term success and patient satisfaction after lower urinary tract reconstruction. MATERIALS AND METHODS: Between May 1988 and November 1996, 28 patients with cervical spinal cord injury underwent lower urinary tract reconstruction. Postoperative information was obtained on 21 patients. Charts were reviewed and patients were contacted by an independent reviewer to ascertain patient satisfaction and quality of life. Patient age was 17 to 51 years (average 34.6). Reconstructive procedures requiring catheterization included augmentation ileocystoplasty in 4 patients plus Mitrofanoff appendicovesicostomy in 7, a Kock ileal reservoir in 8 and an Indiana pouch in 2. RESULTS: Catheterization was regularly performed by 20 of the 21 patients (95%). A total of 12 patients (60%) performed self-intermittent catheterization and 8 (40%) relied on an attendant or family member. Of the patients 80% reported improved quality of life since reconstruction, citing such reasons as lack of a need for urinary drainage bags, increased freedom and an improved sense of body image. Using a visual analog scale to grade satisfaction from 1 to 10 (1-extremely unsatisfied to 10-extremely satisfied) 14 patients (67%) reported a score of 8 or more. CONCLUSIONS: With appropriate preoperative selection of the cervical spinal cord injured patient intermittent catheterization is successfully maintained in the long term, allowing greater flexibility in choice, and a resultant high level of patient satisfaction and improved quality of life. PMID- 12771746 TI - Hautmann and Studer orthotopic neobladders: a contemporary experience. AB - PURPOSE: The development of Studer and Hautmann orthotopic neobladders has revolutionized the management of urinary diversion. Several series have promoted 1 technique over the other. We examine the clinical outcome of a contemporary cohort of patients with bladder cancer who underwent either Hautmann or Studer orthotopic reconstruction. MATERIALS AND METHODS: Retrospective analysis was performed on 130 patients who underwent cystectomy and orthotopic diversion (Studer 93 and Hautmann 37) between March 1, 1995 and September 30, 2000. Clinical parameters evaluated were age, gender, procedure time, length of hospital stay, blood loss, pathological stage, and rates of complication, continence and reoperation. Diversion type was compared with clinical parameters using Fisher's exact test, t-test and Wilcoxon rank sum analysis. RESULTS: Studer and Hautmann techniques had mean procedure times of 5.9 and 5.3 hours, respectively (p = 0.003), and mean length of stay was shorter for Hautmann than Studer (7.0 versus 8.3 days, respectively, p = 0.02). When comparing both populations there was no difference in age, gender, estimated blood loss, pathological stage or rates of complication and reoperation. Total continence was similar in the Hautmann and Studer cohorts, whether during the day (67% versus 67%) or at night (47% versus 40%), respectively. CONCLUSIONS: The data suggest that both orthotopic techniques can be performed in men and women in a safe and timely fashion. Nighttime continence rates are slightly higher with the Hautmann technique, although not statistically different. The shortened Hautmann procedure time likely relates to variations in the ureteral anastomosis. Decreased length of stay may reflect recent trends of early hospital discharge. PMID- 12771747 TI - Enterocystometry: impact of filling rate. AB - PURPOSE: We evaluated different enterocystometric parameters at different filling rates. MATERIALS AND METHODS: A total of 20 male patients who underwent radical cystoprostatectomy and orthotopic detubularized ileal reservoirs with at least 1 year of followup were the material of this study. Enterocystometry was done with a slow fill rate (10 ml. per minute). Under the same circumstances it was then repeated with a medium fill rate (50 ml. per minute). RESULTS: Maximum enterocytometric capacity for slow fill was significantly lower than for medium fill (median 475 versus 610 ml., p <0.001). Volume at first uninhibited contraction for slow fill was significantly lower than for medium fill (median 247 versus 450 ml., p <0.001). Volume at first desire for slow fill was significantly lower than for medium fill (median 306 versus 436 ml., p = 0.012). Moreover, volume at highest contraction for slow fill was significantly lower than for medium fill (median 451 versus 557 ml., p <0.001). Other pressure parameters were comparable without any significant difference between the 2 filling rates. The difference in contraction frequency calculated in the last 100 ml. of filling was significantly higher for slow than for medium fill enterocystometry. CONCLUSIONS: Higher rates of filling during enterocystometry resulted in significantly higher capacity, delayed onset of contractions, delayed onset of first desire (or sense of fullness) and delayed onset of highest contraction. Therefore, we recommend slower filling rates during enterocystometric studies, which mimics physiological states and allows accurate and earlier identification of enterocystometric changes that could be masked by higher rates of filling. PMID- 12771748 TI - Incidence and significance of positive urine cultures in patients with an orthotopic neobladder. AB - PURPOSE: With an intact normal bladder bacterial colonization is uncommon unless intermittent catheterization is instituted. Because intestine, which is normally colonized with bacteria, is used to form an orthotopic neobladder, we determined whether patients with orthotopic urinary diversion are at increased risk for urinary tract infection and urosepsis. MATERIALS AND METHODS: A total of 66 patients who received an orthotopic neobladder after radical cystectomy were prospectively evaluated with urinalysis and culture 2 months to 4 years postoperatively. No patient was on suppressive antibiotics unless they had recurrent urinary tract infections. RESULTS: A total of 55 voided normally and 11 performed intermittent catheterization at least once daily due to high post-void residual urine. Of the patients who voided normally 78% had at least 1 positive urinalysis. If a patient had a positive urinalysis, bacteria was identified on culture in 50%. Overall 26 (39%) and 8 (12%) patients had a urinary tract infection and urosepsis, respectively. The estimated 5-year probability of urinary tract infection and urosepsis for patients who voided independently were 58% and 18%, respectively. Urine culture with greater than 100,000 cfu bacteria and female gender were the only factors predictive of urinary tract infection on multivariate analysis. Recurrent urinary tract infection was the only predictor for urosepsis. Intermittent catheterization or hydronephrosis was not related to urinary tract infection or urosepsis. CONCLUSIONS: The presence of small bowel intestine appears to promote asymptomatic bacterial colonization but urosepsis rarely occurs unless the patient has recurrent urinary tract infections. Prophylactic antibiotics are recommended only for patients with recurring urinary tract infections but treating a positive urinary culture in the absence of specific voiding symptoms is not advocated in this patient population. PMID- 12771749 TI - Treatment of radiation induced hemorrhagic cystitis with hyperbaric oxygen. AB - PURPOSE: Hemorrhagic cystitis can occur 6 months to 10 years after pelvic radiation therapy with moderate to severe persistent rates of hematuria as 3% to 5% after radiotherapy for pelvic malignancies. Current treatment modalities for hemorrhagic cystitis include oral and intravenous agents, intravesical therapy and selective embolization of the hypogastric arteries. Hyperbaric oxygen therapy is now a widely accepted treatment option for radiation induced hemorrhagic cystitis. We assess the efficacy of hyperbaric oxygen for treatment of hemorrhagic cystitis. MATERIAL AND METHODS: From May 1988 through March 2001, 62 patients with radiation induced hemorrhagic cystitis were treated with hyperbaric oxygen at our institution. Followup ranged from 10 to 120 months. The primary pathological conditions were prostate cancer (81%) and bladder cancer (10%). Mean patient age was 70 years (range 15 to 88). Mean time between completion of radiation therapy and onset of hematuria was 48 months (range 0 to 355). Patients received an average of 33 hyperbaric oxygen treatments (range 9 to 68). RESULTS: Of the 62 patients treated information on 57 was available for analysis. Of the 57 patients (86%) 49 experienced complete resolution or marked improvement of hematuria following hyperbaric oxygen treatment. Of the 8 patients who did not improve 4 received fewer than 40 hyperbaric oxygen treatments and 7 prematurely terminated treatment (medical co-morbidities 4, claustrophobia 2, temporary resolution of symptoms 1). CONCLUSIONS: Hyperbaric oxygen therapy for radiation induced hemorrhagic cystitis is an efficacious treatment modality for patients in whom other forms of management have failed. PMID- 12771750 TI - Bullous lesion in the prostatic urethra: morphological change caused by putative chlamydial infection. AB - PURPOSE: The cause of benign bullous lesions in the prostatic urethra, which we encountered in 10 patients during the last 18 years and which has not been described in literature, was studied. MATERIALS AND METHODS: Among 1,236 patients who underwent cystourethroscopy for a urological complaint 10 had bullous lesions in the prostatic urethra which were empirically thought to be inflammatory rather than tumorous lesions at initial cystourethroscopy. We retrospectively searched for common clinical characteristics for these 10 patients who had a median age of 33.5 years (range 20 to 47). The reasons for cystourethroscopy were terminal gross hematuria in 3 patients, initial gross hematuria in 1, total gross hematuria in 2, blood stain on underpants without scrotal keratoangioma in 1, hemospermia in 1 and voiding difficulty in 2. RESULTS: Based on the results of urinalysis, expressed prostatic secretion, transrectal examination of the prostate, cystourethroscopy, urethral swab test for Chlamydia trachomatis and punch biopsy of urethral mucosa with or without immunofluorescence staining with fluorescein isothiocyanate labeled monoclonal antibody for C. trachomatis, inflammation of the prostatic urethra was considered the cause of these bullous lesions. CONCLUSIONS: Bullous lesions in the prostatic urethra appear to be due to an inflammatory change. We should consider these lesions when we encounter young patients with asymptomatic gross hematuria. We should also note microscopic pyuria in such patients and subsequently perform C. trachomatis polymerase chain reaction test using urine initially as a noninvasive examination before cystourethroscopy. PMID- 12771751 TI - Penile calciphylaxis: analysis of risk factors and mortality. AB - PURPOSE: Penile calciphylaxis is a rare condition resulting in infection and gangrene. Most cases are associated with systemic calciphylaxis. The pathophysiology, diagnosis and management of penile calciphylaxis as a distinct entity have received little attention. We reviewed the literature to increase understanding of this disease. MATERIALS AND METHODS: A retrospective review of the literature was performed after treating a case of penile calciphylaxis. Patient characteristics, presentation, serum chemistry studies, management and outcomes are reported. RESULTS: A total of 34 cases of penile calciphylaxis were identified in the literature including our patient. Average patient age was 58 years. All patients had end stage renal disease, and diabetes mellitus was a co morbidity in 76%. Additional areas of gangrene beyond the genitalia were found in two-thirds of patients. Average calcium phosphate product was 78.5 mg.2/dl.2 (range 20.6 to 52.5) and mean parathormone level was 553 pg./ml. (10 to 65). Parathyroidectomy was performed in 8 patients. All patients were treated with either local debridement/wound care or partial/total penectomy. Survival was better in patients who underwent parathyroidectomy (75%) than in those treated with local debridement or penectomy alone (28%). The overall mortality associated with this disease was 64% with a mean time to death of 2.5 months. CONCLUSIONS: Penile calciphylaxis is a result of medial calcification and fibrosis of blood vessels. The co-morbidity and mortality associated with this disease are extremely high. Secondary hyperparathyroidism and an increased calcium phosphate are characteristic and require aggressive medical management. Surgical management of penile lesions and parathormone is controversial. Our review suggests that parathyroidectomy may improve survival and that survival is independent of the type of local treatment for the penile lesions. PMID- 12771752 TI - Urodynamic effect of acute transcutaneous posterior tibial nerve stimulation in overactive bladder. AB - PURPOSE: Of the various treatments proposed for urge incontinence, frequency and urgency electrostimulation has been widely tested. Different techniques have been used with the necessity of surgical implantation (S3 neuromodulation or sacral root stimulation) or without requiring surgery (perineal transcutaneous electrostimulation). Recently peripheral electrical stimulation of the posterior tibial nerve was proposed for irritative symptoms in first intention or for intractable incontinence. Clinical studies have demonstrated good results and urodynamic parameters were improved after chronic treatment. However, to our knowledge no data concerning acute stimulation and immediate cystometry modifications have been reported. We verified urodynamic changes during acute posterior tibial nerve stimulation. MATERIALS AND METHODS: A total of 44 consecutive patients with urge incontinence, frequency and urgency secondary to overactive bladder were studied. There were 29 women and 15 men with a mean age +/-SD of 53.3 +/- 18.2 years. Of the patients 37 had detrusor hyperreflexia due to multiple sclerosis (13), spinal cord injury (15) or Parkinson's disease (9), and 7 had idiopathic detrusor instability. Routine cystometry at 50 ml. per minute was done to select the patients with involuntary detrusor contractions appearing before 400 ml. maximum filling volume. Repeat cystometry was performed immediately after the first study during left posterior tibial nerve stimulation using a surface self-adhesive electrode on the ankle skin behind the internal malleolus with shocks in continuous mode at 10 Hz. frequency and 200 milliseconds wide. Volume comparison was done at the first involuntary detrusor contraction and at maximum cystometric capacity. The test was considered positive if volume at the first involuntary detrusor contraction and/or at maximum cystometric capacity increased 100 ml. or 50% during stimulation in compared with standard cystometry volumes. RESULTS: Mean first involuntary detrusor contraction volume on standard cystometry was 162.9 +/- 96.4 ml. and it was 232.1 +/- 115.3 ml. during posterior tibial nerve stimulation. Mean maximum cystometric capacity on standard cystometry was 221 +/- 129.5 ml. and it was 277.4 +/- 117.9 ml. during stimulation. Posterior tibial nerve stimulation was associated with significant improvement in first involuntary detrusor contraction volume (p <0.0001) and significant improvement in maximum cystometric capacity (p <0.0001). The test was considered positive in 22 of the 44 patients. CONCLUSIONS: These results suggest an objective acute effect of posterior tibial nerve stimulation on urodynamic parameters. Improved bladder overactivity is an encouraging argument to propose posterior tibial nerve stimulation as a noninvasive treatment modality in clinical practice. PMID- 12771753 TI - Effects of ejaculation by penile vibratory stimulation on bladder capacity in men with spinal cord lesions. AB - PURPOSE: We examined the effects of ejaculation by penile vibratory stimulation on bladder capacity in men with spinal cord lesions. MATERIAL AND METHODS: Included in our study were 14 men with spinal cord lesions from C4 to T7 with detrusor hyperreflexia. Cystometry was performed before and immediately after ejaculation by penile vibratory stimulation to establish baseline conditions and repeated after 1 month of ejaculation by penile vibratory stimulation every third day. The third cystometry study was done after 1 month of ejaculation by penile vibratory stimulation every third day at home to determine any long-term effects of treatment. This third cystometry was performed 72 hours after the last ejaculation to exclude any acute effects of ejaculation by penile vibratory stimulation on detrusor hyperreflexia. In addition, 1 to 3 days later ejaculation was induced by penile vibratory stimulation and immediately followed by cystometry to examine whether it was possible to achieve an acute effect as well as a potential long-term effect. RESULTS: Baseline urodynamic investigations revealed bladder hyperreflexia and external sphincter dyssynergia in all individuals. There was no statistically significant difference in bladder capacity at leak point before and immediately after ejaculation by penile vibratory stimulation. However, after 4 weeks of frequent penile vibratory stimulation treatment bladder capacity at leak point increased significantly from a median of 190 ml. (range 17 to 700) at baseline to 293 (range 30 to 700) (Wilcoxon signed rank test p = 0.03). Furthermore, there was a trend toward decreased intravesical pressure during the filling phase. CONCLUSIONS: Ejaculation by penile vibratory stimulation was associated with a significant increase in bladder capacity at leak point after 4 weeks of frequent treatment. This finding may have implications in the management of incontinence in men with spinal cord lesions. PMID- 12771754 TI - A longitudinal assessment of bowel related symptoms and fecal incontinence following radical perineal prostatectomy. AB - PURPOSE: Recent studies have suggested an increased incidence of fecal incontinence following radical perineal prostatectomy. We provide a prospective and longitudinal assessment of bowel related symptoms of patients undergoing radical perineal prostatectomy. MATERIALS AND METHODS: A total of 78 patients who underwent radical perineal prostatectomy between January 1 and December 31, 2001 and had a minimal followup of 6 months were included in the analysis. Patient information was obtained from the chart and the bowel domain specific questions of a validated quality of life questionnaire, the Expanded Prostate Cancer Index Composite. The questionnaire was administered to the candidates preoperatively, at 4 weeks following surgery and subsequently at 3-months intervals. A mean bowel function, bother and summary health related quality of life score was calculated at each interval. The duration of new or worsened symptoms with respect to baseline was evaluated using Kaplan-Meier analysis. RESULTS: Symptoms of involuntary stool leakage and rectal urgency were reported by 11.5% (9 of 78) and 19.2% (15) of patients preoperatively. While all bowel related symptoms transiently increased following surgery, rectal urgency was the most persistent symptom, yet normalized in more than 90% of patients within 9 1/2 months. Compared to individual baseline 15.4%, 7.7%, 5.1% and 3.9% of patients reported worsened symptoms of fecal incontinence after 3, 6, 9 and 12 months, respectively. In the subset of 69 patients who denied preoperative fecal incontinence the incidence of involuntary stool leakage was 2.9% by 12 months following radical perineal prostatectomy. Of 10 patients 9 recovered individual health related quality of life score by 6 months after prostatectomy. CONCLUSIONS: Longitudinal assessment of self-reported questionnaire data suggests that fecal incontinence and bowel related symptoms are more prevalent following radical perineal prostatectomy compared to baseline, yet resolve in the majority of patients with time in the early postoperative period. PMID- 12771755 TI - Intraoperative nerve stimulation with measurement of urethral sphincter pressure changes during radical retropubic prostatectomy: a feasibility study. AB - PURPOSE: We evaluated the feasibility of using intraoperative nerve stimulation and real-time urodynamic monitoring to identify the intrapelvic innervation of the urethral sphincter during radical retropubic prostatectomy. MATERIALS AND METHODS: Using an intraurethral balloon pressure transducer and nerve stimulator changes in urethral pressure were measured in response to stimulation of the neurovascular bundles, pelvic side wall, bladder neck, rectus muscle and other structures in 8 patients undergoing nerve sparing radical retropubic prostatectomy. Intraurethral pressure changes were charted on an urodynamic monitor and correlated with the anatomical location of stimulation. RESULTS: Stimulation of the neurovascular bundles resulted in measurable and significant (greater than 10 cm. H(2)O) increases in intraurethral pressure in all 8 patients. The mean pressure increase was 22 cm. H(2)O. Neither control structure, that is the bladder neck or rectus, resulted in pressure changes with stimulation. In 60% of the subjects pelvic side wall stimulation resulted in urethral pressure increases, while in 40% this stimulation caused pelvic contraction floor but no pressure increase. The mean pressure changes with side wall stimulation was 14 cm. H(2)O. CONCLUSIONS: Intraoperative stimulation of pelvic neural structures and measurement of changes in urethral pressure in response to stimulation are feasible during radical retropubic prostatectomy. Stimulating the neurovascular bundle consistently results in significant increases in urethral pressure. The finding of an intrapelvic urethral innervation supports the previously published observation that nerve sparing radical retropubic prostatectomy may result in improved continence postoperatively. PMID- 12771756 TI - Macroplastique implantation system for the treatment of female stress urinary incontinence. AB - PURPOSE: We evaluated the 12-month followup outcome of the Macroplastique (Uroplasty, Minneapolis, Minnesota) implantation system for the treatment of stress urinary incontinence caused by intrinsic sphincter deficiency using objective and subjective measures, including quality of life impact. MATERIALS AND METHODS: A total of 21 consecutive women with a mean age of 47.4 years and a mean body mass index of 25.65 kg./m.2 in whom intrinsic sphincter deficiency was urodynamically diagnosed were enrolled in the study. Patients were preoperatively assessed by physical examination, quality of life questionnaire, Stamey grading of incontinence, pad use, pad weight test and urodynamic testing. Patients underwent periurethral injection under local anesthesia with the Macroplastique implantation system. The mean volume of silicone elastomers injected was 6.3 ml. RESULTS: As assessed by the King health questionnaire, patient quality of life improved in all domains and in most lower urinary tract symptoms. Patient satisfaction and subjective surgeon evaluation were assessed by Stamey incontinence grading. From patient point of view 12 (57.1%) considered themselves cured, 4 (19%) were improved and 5 (23.8%) had failure. According to subjective surgeon grading 8 patients (38.1%) were considered cured 6 (28.6%) were improved and 7 (33.3%) had failure. Pad use decreased from a mean of 4.38 to 1.29 units daily. According to the pad weight test 13 patients (62%) were dry, 4 (19%) were improved and 4 (19%) had failure. Urodynamic testing demonstrated that 8 patients (40%) were dry and 1 (5%) was improved. CONCLUSIONS: The Macroplastique implantation system proved to have an acceptable outcome for patient and surgeon. The procedure can be done with local anesthesia and without cystoscopic guidance. PMID- 12771757 TI - Preoperative urodynamic evaluation may predict voiding dysfunction in women undergoing pubovaginal sling. AB - PURPOSE: We determine which urodynamic parameters can best predict postoperative voiding dysfunction following pubovaginal sling surgery. MATERIALS AND METHODS: The records of 98 consecutive women who had undergone pubovaginal sling surgery with allograft fascia lata between July 1998 and July 2000 were reviewed. Urodynamic and followup data were sufficient for evaluation for 73 patients. Urodynamic and clinical parameters were correlated with urinary retention, time to return of efficient voiding and development of postoperative urgency symptoms. RESULTS: Average time to return of efficient voiding was 3.92 days (median 3). Of 21 women who voided without a detrusor contraction urinary retention developed in 4 (23%) versus 0 of 48 who voided with detrusor contraction (p = 0.007). Urinary retention was defined as the need to perform even occasional self catheterization. All 4 women with urinary retention had a detrusor pressure of less than 12 cm. H(2)O (0 in 3, 4 in 1). None of the women with a detrusor pressure of greater than 12 cm. H(2)O had urinary retention (p = 0.047). The presence of Valsalva voiding in women without a detrusor contraction did not affect the incidence of urinary retention (11.1%) compared to those who did not demonstrate Valsalva voiding (5.1%) (p = 0.603). Peak flow rate, detrusor instability on preoperative urodynamics and post-void residual urine volume were not associated with postoperative urinary retention. Finally, post-void residual urine volume predicted delayed return to normal voiding (p = 0.001). There were no other urodynamic parameters that were significantly associated with urinary retention, delayed return to normal voiding or postoperative urgency symptoms including peak flow rate, capacity or compliance. CONCLUSIONS: Women who void without or with a weak detrusor contraction are most likely to have urinary retention postoperatively. Therefore, we conclude that preoperative urodynamic evaluation may be used to counsel women regarding the risk of urinary retention following the pubovaginal sling procedure. PMID- 12771758 TI - Transvaginal tape lysis for urinary obstruction after tension-free vaginal tape placement. AB - PURPOSE: We examine the outcome of tape lysis in subjects with obstructive urinary retention after initial tension-free vaginal tape placement. MATERIALS AND METHODS: Charts from 109 consecutive subjects who underwent tension-free vaginal tape placement from 1998 to 2001 were retrospectively reviewed. Of the subjects 78 (72%) consented to participation and completion of a questionnaire. All subjects underwent a routine preoperative evaluation before surgery and followup was implemented through 2002. Tape lysis using a standard technique was performed in 5 patients with documented obstructive urinary retention. Subjective assessment by the validated Bristol Lower Urinary Tract Symptoms questionnaire and a postoperative survey characterized perceptions of incontinence, results of initial tension-free vaginal tape placement and outcome of subsequent tape lysis. RESULTS: The objective incontinence cure rate was 89%. Of the 78 patients 17 (22%) and 21 (27%) experienced transience in either dysuria or incomplete bladder emptying, respectively. Transection for obstructive urinary retention was successfully accomplished in 5 subjects with 4 (80%) remaining continent. CONCLUSIONS: Tape lysis is simple and effective for relieving obstruction, and is not associated with a high rate of recurrent stress urinary incontinence. PMID- 12771759 TI - Management of vaginal erosion of polypropylene mesh slings. AB - PURPOSE: The SPARC (American Medical Systems, Minneapolis, Minnesota) polypropylene sling has recently been introduced as an alternative delivery system to TVT (Ethicon, New Brunswick, New Jersey) tension-free vaginal tape for placement of a tension-free mid urethral sling. Erosion must always be considered a risk of synthetic materials. We present 4 cases of vaginal erosion of polypropylene mesh placed with this system and the successful conservative management done. MATERIALS AND METHODS: A total of 90 patients received a SPARC polypropylene pubovaginal sling at our institution between October 1, 2001 and October 1, 2002. During followup 3 of our patients and 1 patient with tension free vaginal tape who was referred from elsewhere presented with vaginal exposure of the mesh. RESULTS: Two patients described persistent vaginal discharge 6 weeks postoperatively, including 1 who complained primarily of partner discomfort during sexual intercourse. Two patients were completely asymptomatic and mesh erosion was discovered at routine physical examination 6 weeks postoperatively. Pelvic examination demonstrated vaginal exposure of the mesh in all cases. Each patient was observed conservatively and 3 months postoperatively all 4 had complete spontaneous epithelialization over the mesh. None had stress incontinence, urgency or urge incontinence, all emptied the bladder to completion and all patients were completely satisfied with the procedure. CONCLUSIONS: The recent literature suggests that polypropylene mesh erosion should be treated with complete removal of the sling material. We present 4 cases of vaginal erosion of polypropylene slings that were managed conservatively with observation and resulted in complete spontaneous healing. Sling preservation with continued patient continence and satisfaction is a feasible option in those with vaginal exposure of polypropylene mesh. PMID- 12771760 TI - Bone anchor 4-corner cystourethropexy: long-term results. AB - PURPOSE: We evaluate the long-term results of incision less transvaginal bone anchor cystourethropexy to treat genuine urinary stress incontinence. MATERIALS AND METHODS: Between August 1995 and January 1997, 31 women 36 to 81 years old (mean age 58) with types I and II genuine urinary stress incontinence were treated. Four miniature bone anchors (2 on each side of the urethra) attached to a suture were fired transvaginally into the retropubic bone using a bone anchor stapler. The ipsilateral sutures were tied, thus creating a Marshall-Marchetti colposuspension. RESULTS: Of the patients 3 were lost to followup and the remaining 28 were followed for at least 60 months. Only 6 patients (21.4%) are continent. In 5 patients 11 sutures passed through the bladder 5 of which were removed intraoperatively and the other 6 were detected and removed during followup. Incontinence recurred in 1 patient because the sutures cut through the vaginal tissue and were found loose in the retropubic space. In 8 patients 12 anchors had become detached from the bone of which 7 were detected on x-ray in the retropubic area, 2 were removed cystoscopically 1 year later and 3 were spontaneously expulsed through the vagina 10 months to 5 years postoperatively. In 1 patient with intravesical sutures a vesicovaginal fistula developed which was successfully repaired, and in another pubic osteomyelitis developed. CONCLUSIONS: We regard 4-corner bone anchor cystourethropexy as unsuitable for genuine urinary stress incontinence based on the unfavorable outcome of incontinence and high rate of complications, including a relatively high incidence of confirmed anchor detachment. PMID- 12771761 TI - Outcome of urethral reconstructive surgery in a series of 74 women. AB - PURPOSE: We evaluated the results of vaginal flap reconstructive surgery of the female urethra. MATERIALS AND METHODS: A series of 74 consecutive women who presented with urethral pathology requiring surgical reconstruction were assessed for anatomical and functional outcome. Followup was 1 to 15 years (median 1.5). Outcome assessment included success or failure of the anatomical repair based on physical examination as well as voiding habits and continence assessed by voiding diary, questionnaire and patient self-assessment. RESULTS: A total of 74 procedures were performed, including 72 vaginal flap repairs (56 with a concomitant pubovaginal sling). Five vaginal flap repairs were performed with a concomitant modified Pereyra procedure and 1 was done with Kelly plication. A patient in whom vaginal flap repair was planned underwent a Tanagho anterior bladder flap procedure, 1 underwent extensive lysis of urethral and periurethral scarring with primary urethral repair and 12 underwent vaginal flap repair without an incontinence procedure. A total of 72 women with an average age +/- SD of 54 +/- 13 years were followed a median of 1.5 years after vaginal flap reconstruction and 2 were lost to followup. Of the 62 patients with preoperative incontinence 54 (87%) considered themselves cured or improved. Successful anatomical repair by single procedure vaginal flap repair was achieved in all except 5 patients (93%). CONCLUSIONS: Single stage vaginal flap reconstruction with concurrent pubovaginal sling and Martius flap graft has a high degree of anatomical and functional success for treating a difficult surgical problem. PMID- 12771762 TI - Pregnancy outcomes after vasectomy reversal for female partners 35 years old or older. AB - PURPOSE: We review the outcomes after vasectomy reversal for couples with female partners 35 years old or older. MATERIALS AND METHODS: A retrospective review of experience at 2 institutions was performed. Patency was defined as the presence of motile sperm. Patients with less than 6 months of followup were excluded from the patency rate analysis unless they had sperm in the semen. Similarly, patients with less than 12 months of followup or no ongoing interest in establishing conception were excluded from the pregnancy rate analysis unless they had established a pregnancy or they were azoospermic with sufficient followup. RESULTS: A total of 46 men with partners 35 years old or older underwent vasectomy reversal at 2 institutions. Mean partner age was 37 +/- 2 years, and median obstructive interval was 10 years. Bilateral vasovasostomy was performed in 43 men, unilateral vasovasostomy in 2 and vasovasostomy/vasoepididymostomy in 1. Of the 46 men 27 had followup semen analyses with a patency rate of 81% (22). Transient patency occurred in 2 cases (7%). Pregnancy occurred in 35% of the couples (14 of 40 patients) with sufficient followup. The ongoing/live delivery rate was 33% (13 of 40 cases). The pregnancy and ongoing/delivery rates were 46% (12 of 26 patients) and 46% (12 of 26) for female partners 35 to 39 years old, and 14% (2 of 14) and 7% (1 of 14) for female partners older than 40, respectively. CONCLUSIONS: Vasectomy reversal offers reasonable chance for success when the female partner is 35 years old or older. The chance for success is similar to that of a single cycle of in vitro fertilization with intracytoplasmic sperm injection. These couples should not be eliminated from consideration for reversal simply because the female partner is 35 years old or older. PMID- 12771763 TI - Combination treatment with an alpha-blocker plus an anticholinergic for bladder outlet obstruction: a prospective, randomized, controlled study. AB - PURPOSE: We evaluate the effect of tolterodine combined with tamsulosin on quality of life in patients with bladder outlet obstruction and concomitant detrusor instability. MATERIALS AND METHODS: The study included 50 consecutive patients with urodynamically proven mild or moderate bladder outlet obstruction and concomitant detrusor instability. All patients were initially treated with 0.4 mg. tamsulosin orally once a day. A week later the patients were randomly allocated into group 1-25 who continued treatment with tamsulosin only and, group 2-25 who also received 2 mg. tolterodine orally twice daily. Reevaluation with a quality of life questionnaire and urodynamic study was performed after 3 months. RESULTS: Two patients from group 2 stopped tolterodine while 1 patient from each group stopped tamsulosin because of hypotension. Analysis revealed statistically significant improvement in quality of life scores only in group 2 patients (mean score 525.0 and 628.4 before and after treatment, respectively, 2-sided t test p = 0.0003). A significant difference was noted in both groups after treatment for maximum flow rate and volume at first contraction. Additionally, in group 2, a statistically significant difference was observed for maximum detrusor pressure and maximum unstable contraction pressure after treatment. CONCLUSIONS: Combination treatment with an alpha-blocker (tamsulosin) plus an anticholinergic (tolterodine) improves quality of life in patients with bladder outlet obstruction and concomitant detrusor instability. Interestingly, no acute urinary retention was observed and tolterodine did not affect the quality of urine flow or residual urine volume. The proposed combination appears to be an effective and relatively safe treatment option in patients with bladder outlet obstruction and detrusor instability. PMID- 12771764 TI - Sexual dysfunction in 1,274 European men suffering from lower urinary tract symptoms. AB - PURPOSE: We determine the prevalence and bothersomeness of erectile and ejaculatory dysfunction in men with lower urinary tract symptoms suggestive of bladder outlet obstruction in real life practice, and analyze predictors of sexual dysfunction in this population. MATERIALS AND METHODS: Sexual function of 1,274 European men with lower urinary tract symptoms was assessed by the DAN PSSsex questionnaire. The relationship between sexual dysfunction and selected clinical characteristics was analyzed. RESULTS: The proportion of sexually active men decreased from 91% in those 60 years old or younger to 60% in those 70 years old or older. Erectile dysfunction, reduced ejaculation and pain/discomfort on ejaculation were reported by 62%, 63% and 23% of patients, respectively. Erectile dysfunction strongly related with age (40% younger than 60 years, 80% 70 years old or older), lower urinary tract symptom severity (55% mild, 70% severe) and body mass index, hypertension and concomitant treatment with calcium channel antagonists. Reduced ejaculation was significantly related to age (42% younger than 60 years, 82% 70 years old or older), lower urinary tract symptom severity (55% mild, 68% severe) and previous benign prostatic hyperplasia surgery. Pain/discomfort on ejaculation was only related to lower urinary tract symptom severity (7% mild, 31% severe). Erectile dysfunction, reduced ejaculation and pain/discomfort on ejaculation were considered a problem by 96%, 82% and 91% of patients younger than 60 years, respectively. These 3 sexual symptoms were still highly bothersome in patients 70 years old or older. CONCLUSIONS: Erectile dysfunction and reduced ejaculation are highly prevalent in men with lower urinary tract symptoms, and are strongly related to increasing age and lower urinary tract symptom severity. Both these aspects of sexual dysfunction are also highly bothersome, even in advanced age. Sexual function should be carefully assessed in the initial evaluation of patients with lower urinary tract symptoms and in deciding on treatment options, as it may have a negative impact on sex life. PMID- 12771765 TI - Laboratory evaluations of erectile dysfunction: an evidence based approach. AB - PURPOSE: We evaluate the prevalence of laboratory abnormalities in men presenting for initial evaluation and therapy of erectile dysfunction. MATERIALS AND METHODS: The computerized charts of men receiving treatment for erectile dysfunction from 1987 to 2002 were retrospectively reviewed. We pooled laboratory data for 3,547 men with erectile dysfunction to assess the prevalence of laboratory abnormalities. Values of the common laboratory screening tests for erectile dysfunction were recorded for testosterone, prolactin, luteinizing hormone, thyroid-stimulating hormone, hemoglobin A(Ic), prostate specific antigen, hemoglobin, cholesterol and creatinine. RESULTS: Of those patients evaluated 18.7% had low testosterone, 4.6% had increased prolactin, 14.6% had abnormal luteinizing hormone, 4.0% had increased thyroid-stimulating hormone, 8.3% had increased prostate specific antigen, 26.5% had anemia and 11.9% tested had renal insufficiency. A high percentage of patients presenting with a primary complaint of erectile dysfunction had increased hemoglobin A(Ic) and total serum cholesterol levels (52.9% and 48.4%, respectively). CONCLUSIONS: An evidence based approach to standardization of laboratory evaluations for men presenting with erectile dysfunction is recommended. Laboratory screening should be directed to identify those risk factors that may benefit from lifestyle modification and pharmacological intervention. PMID- 12771766 TI - Male sexual dysfunction--how little do we know? PMID- 12771767 TI - Quantitative somatosensory testing of the penis: optimizing the clinical neurological examination. AB - PURPOSE: Quantitative somatosensory testing, including vibration, pressure, spatial perception and thermal thresholds of the penis, has demonstrated neuropathy in patients with a history of erectile dysfunction of all etiologies. We evaluated which measurement of neurological function of the penis was best at predicting erectile dysfunction and examined the impact of location on the penis for quantitative somatosensory testing measurements. MATERIALS AND METHODS: A total of 107 patients were evaluated. All patients were required to complete the erectile function domain of the International Index of Erectile Function (IIEF) questionnaire, of whom 24 had no complaints of erectile dysfunction and scored within the "normal" range on the IIEF. Patients were subsequently tested on ventral middle penile shaft, proximal dorsal midline penile shaft and glans penis (with foreskin retracted) for vibration, pressure, spatial perception, and warm and cold thermal thresholds. RESULTS: Mixed models repeated measures analysis of variance controlling for age, diabetes and hypertension revealed that method of measurement (quantitative somatosensory testing) was predictive of IIEF score (F = 209, df = 4,1315, p <0.001), while site of measurement on the penis was not. To determine the best method of measurement, we used hierarchical regression, which revealed that warm temperature was the best predictor of erectile dysfunction with pseudo R(2) = 0.19, p <0.0007. There was no significant improvement in predicting erectile dysfunction when another test was added. Using 37C and greater as the warm thermal threshold yielded a sensitivity of 88.5%, specificity 70.0% and positive predictive value 85.5%. CONCLUSIONS: Quantitative somatosensory testing using warm thermal threshold measurements taken at the glans penis can be used alone to assess the neurological status of the penis. Warm thermal thresholds alone offer a quick, noninvasive accurate method of evaluating penile neuropathy in an office setting. PMID- 12771768 TI - Blood gas changes in the corpora cavernosa: metabolic and histomorphometric implications in the patient with erectile dysfunction. AB - PURPOSE: We evaluated corpora cavernosa metabolism in flaccidity and in erection, analyzing some blood gas analytical parameters and comparing them by histomorphometric examination to find a direct relation between biochemical metabolic parameters and histological data. MATERIALS AND METHODS: We selected 33 patients with erectile dysfunction and divided them into 2 groups, including 1 those with congenital penile deviation who were responders to prostaglandin E1, and 2-those with severe organic erectile dysfunction who were not responders to prostaglandin E1. We evaluated O(2) and CO(2) pressure, pH and O(2) saturation in blood samples. We then made a histomorphometric study of cavernous tissue. We obtained specimens by cavernous biopsies and calculated O(2) and CO(2) exchange, the Haldane effect and the respiratory quotient into the corpora cavernosa. All data were evaluated by statistical analysis. RESULTS: Mean O(2) arterial pressure and saturation +/- SD were lower in group 2 than in group 1 (74.85 +/- 8.78 versus 96.43 +/- 14.87 and 94.98 +/- 1.4 versus 97.35 +/- 0.83, respectively). Mean CO(2) arterial pressure was 35.59 +/- 4.78 group 1 versus 38.8 +/- 2.71 in group 2 with borderline statistical significance. The Haldane effect was superior in flaccidity than in erection because of the influence of arterial-venous O(2) difference and the respiratory quotient, which was also an inverse ratio. Cavernous histomorphometry showed that in group 1 smooth muscle was a mean of 38.8 +/- 8.94% of cavernous tissue versus 24.9% in group 2. CONCLUSIONS: Our study shows that starting with blood gas analytical data we can completely study the metabolism of the corpora cavernosa and its relationships to erectile dysfunction. Cavernous histomorphometry can suggest the presence of smooth muscle into cavernous tissue, whereas our mathematical elaboration allowed us to evaluate all data in a more complete manner. PMID- 12771769 TI - The influence of paternal age on down syndrome. AB - PURPOSE: Children born to older parents are at greater risk for genetic abnormalities, such as Down syndrome. The influence of maternal age on Down syndrome is well established but little is known about the genetic consequences of advanced paternal age. MATERIALS AND METHODS: Data on the incidence of Down syndrome from 1983 to 1997 (3,419 cases) were obtained from the New York State Department of Health congenital malformations registry. Parental age was modeled as individual age groups and by a single linear covariate (drift model). The log linear chi-square test and a test of significance of different explanatory variables were used to evaluate these models to determine significance. We compared actual Down syndrome rates by maternal age with the estimated rate corrected for paternal age. RESULTS: From 1983 to 1997 a dramatic increase in the number of infants born to parents 35 years or older was observed. During the 15 year study period there was an increase of 111% and 60% in the number of mothers and fathers 35 years old or older, respectively. There was no parental age influence on Down syndrome until age 35 years and older. A paternal age effect was seen in association with a maternal age of 35 years and older, and it was most pronounced when maternal age was 40 years and older (p = 0.0004). In this later maternal age group the paternal contribution to Down syndrome was 50%. CONCLUSIONS: Advanced paternal age combined with maternal age significantly influences the incidence of Down syndrome. This effect may represent a paradigm for other genetic abnormalities in children of older fathers. PMID- 12771770 TI - Regret in men treated for localized prostate cancer. AB - PURPOSE: We identify the predictors of medical regret in men treated for localized prostate cancer. MATERIALS AND METHODS: Patients previously treated for early stage prostate cancer were assessed for treatment regret using validated items. Univariate and multivariate analyses identified associations between regret and demographic characteristics, clinical outcomes, medical knowledge, and general and disease specific health related quality of life as measured by the general health perceptions domain of the RAND 36-Item Health Survey and a validated short form of the University of California, Los Angeles Prostate Cancer Index. RESULTS: Of 96 respondents (mean age 64 years, mean followup 2.8 years) 16% expressed regret with treatment decisions. Regretful men were almost twice as likely as nonregretful men to have less than a college education (60% versus 33%, p = 0.05) and worse current health related quality of life (p <0.05). In addition, regretful men tended to be unable to recall the most recent prostate specific antigen accurately (p = 0.06). Men with and without regret did not differ in other demographic characteristics, treatment choice or clinical outcomes. Regretful men were more likely to say they would choose a different treatment if they could. In multivariate analyses worse quality of life predicted regret but decline in quality of life with time was not associated with regret. CONCLUSIONS: Men expressing regret over treatment choice for localized prostate cancer have poorer health related quality of life. Further study is needed to identify factors that predict posttreatment regret. Such information will allow patients and physicians to individualize treatment decisions, optimize quality of life and avoid medical regret. PMID- 12771771 TI - Management of obliterated urinary segments using a laser fiber for access. AB - PURPOSE: We describe a modification of the cut to the light and core-through procedures using a laser fiber to gain through and through access for treatment of complete occlusion of the ureter or urethra. MATERIALS AND METHODS: Three patients presented with complete obliteration of the ureter (2) and vesicourethral anastomosis (1). In each case the laser fiber was advanced through the obliterated segment under direct vision and then exchanged for a standard guide wire using an open ended catheter. RESULTS: Through and through access was obtained in all 3 cases and allowed subsequent incision for repair. In all 3 patients the area of incision was stented and urinary continuity was restored. CONCLUSIONS: Loss of access can lead to complications and termination of any endoscopic procedure. Use of the laser fiber to incise through the obliterated segment and subsequently act as a guide wire in our experience minimized the possibility for loss of access. The disadvantage is the cost of the laser fiber, which cannot be reused. Use of a laser fiber as a guide wire can be a viable and effective option for gaining access across strictures when alternative methods fail. PMID- 12771772 TI - Novel technique of renal entrapment for morcellation. AB - PURPOSE: Renal entrapment for morcellation can be a difficult task, often necessitating additional port placement and awkward maneuvering as well as concern with specimen sack breakage. We describe a novel technique that combines the best features of existing technology to allow easy entrapment and safe morcellation. MATERIALS AND METHODS: We combined a 15 mm. EndoCatch II (United States Surgical Corp., Norwalk, Connecticut) introducer and frame with an 8 x 10 inch LapSac (Cook Urological, Spencer, Indiana) to facilitate entrapment with a strong reliable specimen sack. The LapSac is sutured in place onto the frame of the EndoCatch device with 3-zero sutures. The device is then rearmed and lubricated prior to introduction. RESULTS: The described modification greatly enhances our ability more rapidly and easily to entrap nephrectomy specimens prior to tissue morcellation. No additional ports are needed and no morbidity has resulted from the use of the modified entrapment device. Introduction and deployment of the modified entrapment device is straightforward and easily reproducible. CONCLUSIONS: We recommend this easy and secure technique when tissue morcellation is being considered. PMID- 12771773 TI - Vattikuti Institute prostatectomy: technique. AB - PURPOSE: We have performed more than 250 radical prostatectomies using the da Vinci (Intuitive Surgical, Mountain View, California) surgical system. Our initial cases were done using the classic Montsouris approach. However, after gaining familiarity with the robot we modified our technique to reflect our experience with open radical retropubic prostatectomy. We detail the Vattikuti Institute prostatectomy technique that we currently use. MATERIALS AND METHODS: The robotic technique requires 2 teams, namely a skilled laparoscopic team at the patient and a skilled open surgeon at the console. Dissection is started anterior to the bladder and it continues extraperitoneally. The endopelvic fascia is opened and the dorsal vein complex is secured. The apex of the prostate is dissected free, releasing the neurovascular bundles at the apex. The bladder neck is then incised, and the seminal vesicles and vasa are transected. Posterior dissection is done within the posterior layer of Denonvilliers' fascia, preserving the neurovascular bundles and lateral prostatic fascia. The apex is transected and frozen sections are obtained from the parietal margins. Vesicourethral anastomosis is formed with 2 continuous sutures. RESULTS: In the last 100 cases mean operative time was 2.5 hours and average blood loss was 150 ml. (range 25 to 525 cc.). Median specimen Gleason score was 7 and mean tumor volume was 7 cc. Four patients had a positive surgical margin, which was focal in 3. Of the patients 95% were discharged home within 23 hours. Mean catheterization time was 4.2 days. CONCLUSIONS: Vattikuti Institute prostatectomy is a precise and safe minimally invasive technique of radical retropubic prostatectomy. PMID- 12771775 TI - Laparoscopic distal ureterectomy for low grade transitional cell carcinoma. PMID- 12771774 TI - The nutcracker syndrome. PMID- 12771776 TI - Renal cell carcinoma with tumor thrombus extension into the proximal pulmonary artery. PMID- 12771777 TI - Unusual mass of the spermatic cord. PMID- 12771778 TI - Incisional bladder hernia after rectus fascial sling. PMID- 12771779 TI - Vaginal granulation tissue secondary to bone anchors: experience in two patients. PMID- 12771780 TI - Priapism following ingestion of tamsulosin. PMID- 12771781 TI - Re: Chromophobe renal cell carcinoma in a patient with the Birt-Hogg-Dube syndrome. PMID- 12771782 TI - Re: Outcome of endoscopic treatment for vesicoureteral reflux in children using polydimethylsiloxane. PMID- 12771783 TI - Re: Effect of nonsteroidal anti-inflammatory agents and finasteride on prostate cancer risk. PMID- 12771784 TI - Re: Chromogranin A concentration as a serum marker to predict prognosis after endocrine therapy for prostate cancer. PMID- 12771785 TI - Re: Lymphatic mapping and gamma probe guided laparoscopic biopsy of sentinel lymph node in patients with clinical stage I testicular tumor. PMID- 12771786 TI - Re: Androgen imprinting of the brain in animal models and humans with intersex disorders: review and recommendations. PMID- 12771787 TI - The role of gadolinium enhanced magnetic resonance imaging for children with suspected acute pyelonephritis. AB - PURPOSE: The diagnosis of pyelonephritis is primarily clinical. However, the history and physical findings can be confusing in children, leading to adjunctive nuclear renal cortical scintigraphic studies (99mtechnetium dimercapto-succinic acid [DMSA]) to confirm the diagnosis. Nonetheless, ambiguity occurs when differentiating between acute pyelonephritis and chronic scarring. We report our initial experience with gadolinium enhanced inversion recovery magnetic resonance imaging (MRI) to diagnose acute pyelonephritis. MATERIALS AND METHODS: Nine patients 7 months to 18 years old (mean age 81 months) underwent MRI to confirm radiographically a clinical suspicion of acute pyelonephritis. All patients had at least 1 prior episode of clinical pyelonephritis. Data were collected to determine whether acute pyelonephritic changes could be differentiated from chronic pyelonephritis on the basis of MRI characteristics. RESULTS: Of the 9 patients 4 were identified as having acute pyelonephritis on MRI (persistently high signal intensity after gadolinium), 2 demonstrated evidence of postpyelonephritic scar (parenchymal loss without change in signal intensity), 1 had evidence of acute pyelonephritis and chronic changes, and 2 had a completely normal examination (decreased signal intensity after gadolinium). At our institution the billable cost of MRI to the patient is $1,329, while the billable cost of 99mtechnetium DMSA is $1,459. All patients younger than 6 years required intravenous sedation for MRI, whereas 70% of those younger than 6 years require intravenous sedation for DMSA scanning at our institution. MRI provided greater anatomical detail regarding the renal architecture without radiation exposure, and allowed the unambiguous diagnosis of acute versus chronic pyelonephritis scar in a 1-time (versus often multipart for DMSA) imaging study. CONCLUSIONS: In cases where adjunctive imaging studies are useful to make a diagnosis gadolinium enhanced inversion recovery magnetic resonance imaging allows the detection of acute pyelonephritis rapidly, cost-effectively and safely in the pediatric population. PMID- 12771788 TI - Measuring glomerular filtration rate with cystatin C and beta-trace protein in children with spina bifida. AB - PURPOSE: We compared the diagnostic performance of cystatin C and beta-trace protein with serum creatinine and the Schwartz formula for the estimation of glomerular filtration rate in children with spina bifida. MATERIALS AND METHODS: Cystatin C, beta-trace protein and serum creatinine in children who underwent routine measurement of glomerular filtration rate were obtained in a prospective study. A spina bifida group (27 measurements in 27 patients 1.4 to 20.0 years old, 15 female, 12 male) was compared with a group of controls (211 measurements in 201 patients with various other renal pathologies 1.0 to 20.2 years old, 89 female, 122 male). All patients underwent nuclear medicine clearance investigations with 99mtechnetium diethylenetetraminepentaacetic acid. Z scores were calculated for body mass index, height and weight. Receiver operating characteristic (ROC) analysis and ROC plots were done to assess their diagnostic accuracy. RESULTS: Serum creatinine and the Schwartz formula did not correlate with glomerular filtration rate in the patients with spina bifida. Correlation coefficients for cystatin C were 0.45 and 0.84 in the spina bifida and control groups, respectively, while correlation coefficients for beta-trace protein were 0.31 and 0.75, respectively. For both groups ROC area was highest for cystatin C (up to 0.97 ROC area in control group). CONCLUSIONS: Only cystatin C served as a suitable marker to estimate glomerular filtration rate in patients with spina bifida. Cystatin C proved to be a superior marker in patients with spina bifida. In the control group the Schwartz formula was comparable to cystatin C and beta trace protein, although cystatin C remained superior. PMID- 12771790 TI - Modified cecal flap neoappendix for the Malone antegrade continence enema procedure: a novel technique. AB - PURPOSE: We report our experience with the creation of a tubularized cecal flap to construct a neoappendix. This flap effectively serves as an alternative means of creating a conduit for antegrade continence enemas in patients who have either an absent or an unusable appendix. MATERIALS AND METHODS: We reviewed our results from our initial 4 patients in whom we used this technique. A neoappendix is created from a medially based flap of distal cecum that is tubularized over a catheter and then buried in a seromuscular tunnel to create an antireflux mechanism. A skin stoma is then created. RESULTS: All stomas are continent, functional and without any evidence of stenosis or necrosis of the cecal flap with followup ranging from 3 months to 8 years. CONCLUSIONS: A medially based cecal flap neoappendix is an easily created and reliable conduit for antegrade enemas. This procedure should be considered within the surgical armamentarium of urologists and pediatric surgeons when the appendix is absent, atretic or fibrotic. PMID- 12771789 TI - Aerosol transfer of bladder urothelial and smooth muscle cells onto demucosalized colonic segments: a pilot study. AB - PURPOSE: We developed a cell transfer technology for covering demucosalized colonic segments with bladder urothelium. This covering would be achieved through aerosol spraying of single cell suspension of bladder urothelial and smooth muscle cells with fibrin glue onto the demucosalized colonic segments. MATERIALS AND METHODS: In 6 piglets (20 kg.) a 4 cm.2 area of bladder was excised. Single cell suspension of bladder urothelial and smooth muscle cells was prepared. A segment of detubularized sigmoid colon was isolated on its vascular pedicle and demucosalized. The single cell suspensions were combined with an equal volume of fibrin glue and sprayed over the raw submucosal surface of the sigmoid segment. The sigmoid segment was retubularized and sutured to the posterior peritoneum. Animals were sacrificed 4 weeks later, and the segment was submitted to histological and immunohistochemical analysis. RESULTS: Sigmoid segments appeared grossly intact with no reduction in surface area. Hematoxylin and eosin architecture revealed an intact urothelial layer. Deep to this layer was a randomly aligned but distinctly segregated layer of smooth muscle cells. The urological new smooth muscle layer stained positive for calponin and the urothelial layer was cytokeratin-7 and uroplakin III positive. CONCLUSIONS: Separation, cell suspension and aerosol delivery of bladder urothelial and smooth muscle cells in fibrin glue can successfully transfer these urological cell populations to a new host tissue commonly used in urological reconstruction. In vivo co-culture of bladder smooth muscle and urothelial cells results in coverage of a large area of demucosalized gut providing new potential for transfer and reconstitution of urologically functionally appropriate tissue to the bladder itself. PMID- 12771791 TI - Mathieu urethroplasty as a salvage procedure: 20-year experience. AB - PURPOSE: We assessed the results of the Mathieu procedure in a series of patients who had undergone multiple prior hypospadias repairs. MATERIALS AND METHODS: Medical records of 55 boys who underwent repeat surgery with the Mathieu procedure were retrospectively reviewed. Prior surgeries, level of hypospadias defect, number of prior surgeries and complications were recorded. Statistical analyses were performed on factors that would impact the outcome. RESULTS: Mean patient age at the time of the salvage Mathieu procedure was 10.33 years (range 2 to 30). In 14 of the 55 patients (25.4%) fistula occurred following the salvage repair. Complications were split glans in 1 case, skin dehiscence in 1 and complete breakdown of the repair in 1. Fistula formation did not differ in groups categorized according to the level of the urethral meatus. Fistula occurred in 6 of 37 (16.2%) and 8 of 18 (44.4%) boys who underwent 1 and more than 1 prior surgery, respectively (p = 0.04). CONCLUSIONS: Although the Mathieu operation had a favorable outcome (74.6% cure) as a salvage procedure in 1 stage, fistula occurred more frequently as the number of prior surgeries increased. Therefore, if primary surgery fails, repeat operation should be performed by an experienced "hypospadiologist." PMID- 12771792 TI - Prospective evaluation of human chorionic gonadotropin in the differentiation of undescended testes from retractile testes. AB - PURPOSE: We prospectively evaluated the efficacy of human chorionic gonadotropin (HCG) in the treatment of undescended testis and sought to determine whether HCG assists in the differentiation of undescended testis from retractile testis. MATERIALS AND METHODS: Patients with undescended testes were offered HCG. Testis position, laterality and the presence or absence of a hypoplastic scrotum were noted. The same physician (G. W. K.) recorded physical findings prospectively and stated clinical impression of descent. RESULTS: A total of 67 patients with 90 undescended or retractile testes were treated and evaluated with HCG. Of the 64 undescended testes 13 (20%) descended with HCG therapy, with none requiring subsequent surgery. Of the 26 retractile testes 15 (58%) descended with HCG (p <0.001). Based on physical examination, 100% of retractile testes descended if the testis was in the high scrotal position but only 40% descended if the testis was in the superficial pouch or inguinal area. In the undescended testes group no ectopic or nonpalpable testis descended with HCG. Evaluation of HCG with age demonstrated minimal response (15%) to HCG at less than 24 months, and a peak response between ages 2 and 6 years (75%) with response decreasing thereafter. CONCLUSIONS: HCG may have a limited role in the evaluation of undescended testis in patients younger than 2 years. HCG can serve as an adjunct in the clinical diagnosis of retractile testis in older children. PMID- 12771793 TI - Outpatient management of phimosis following newborn circumcision. AB - PURPOSE: We reviewed our experience in treating patients with a trapped penis due to phimosis following newborn circumcision. The outcome of treatment of this condition at the outpatient clinic was examined. Possible etiological factors leading to this complication were determined. METHODS: A total of 521 pediatric patients underwent Gomco circumcision while under local anesthesia at our outpatient clinic between 1994 and 1999. Phimosis was noted in 15 patients (2.9%), and an additional 13 patients were referred to our clinic with phimosis after undergoing circumcision elsewhere. These 28 patients underwent treatment for the phimosis at the outpatient clinic. The phimotic ring was manually dilated with a fine hemostat. Careful traction with sterile gauze was then used to reduce the phimotic band below the glans penis. No evidence of glans ischemia was noted. Mild edema of the pericoronal skin collar was common. Cases referred after age 6 months or those weighing more than 14 pounds were treated as an elective outpatient surgical procedure. RESULTS: Three patients had recurrence of the phimosis. These cases were managed with manual retraction of the prepuce at the clinic. At 1 month followup 27 patients had resolution of the phimosis. One infant was lost to followup. Factors contributing to development of phimosis after newborn circumcision were reviewed. Ten of the patients had obvious poor penile skin attachment with concealment of the penis even after resolution of the phimosis. Phimosis occurred more frequently in older patients undergoing circumcision-15 patients were older than 1 month at circumcision. Mean body weight of the latter group at circumcision was 12 pounds. CONCLUSIONS: Phimosis with a trapped penis is an infrequent but important complication of circumcision. This condition is more likely to occur in older infants and those with poor attachment of the penile skin to the shaft. Early recognition allows outpatient treatment with excellent results, avoiding operative intervention with general anesthesia. PMID- 12771794 TI - Pelviureteral junction obstruction: correlation of renal cell apoptosis and differential renal function. AB - PURPOSE: We evaluated the relationship between renal biopsy changes and preoperative and postoperative renal scans in 29 male and 14 female infants with prenatal severe hydronephrosis and unilateral ureteropelvic junction obstruction. We also verified on immunohistochemical studies glomerular changes, degeneration of the epithelium of the proximal tubules, interstitial fibrosis and inflammation and apoptotic nuclei. MATERIALS AND METHODS: In the children, all with prenatal diagnosis of hydronephrosis, ureteropelvic junction obstruction was diagnosed with 99mtechnetium mercaptoacetyltriglycine renal scan performed in all patients at ages 4 to 6 weeks to establish baseline differential renal function. All patients underwent renal biopsies at the time of pyeloplasty. The biopsy samples were examined with histological and immunohistochemical methods for antigens associated with apoptosis, such as clusterin, CD95, TDAG51 and bcl-2. Renograms were performed 3 months after surgical repair. RESULTS: The male-to-female ratio was 2.1:1. There was no difference between males and females in regard to baseline differential renal function of the affected kidney. All biopsy samples confirmed degeneration of the epithelium of the proximal tubules, interstitial focal fibrosis was found in 4 samples, mild chronic inflammation with lymphoid aggregates in 4 and focal Bowman's space dilatation in 1. No specimen demonstrated apoptotic nuclei as confirmed by immunohistochemical study which showed the presence of bcl-2 and absence of CD95, TDAG51 and clusterin, probably because there was no dysplasia in the samples examined. CONCLUSIONS: These results indicate absence of apoptosis in the kidney with preoperative uptake less than 40% and minimal histological changes. PMID- 12771795 TI - The PIC cystogram: a novel approach to identify "occult" vesicoureteral reflux in children with febrile urinary tract infections. AB - PURPOSE: When the standard cystogram does not show vesicoureteral reflux in children who have experienced febrile urinary tract infections (UTIs), clinical management is controversial. We postulated that vesicoureteral reflux accounts for such UTIs but is "occult." We tested this hypothesis by using a novel method, PIC cystography (Positioning the Instillation of Contrast at the ureteral orifice) at the time of cystoscopy. MATERIALS AND METHODS: We performed PIC cystography with instillation of contrast medium at the ureteral orifice consecutively and prospectively in 57 children who underwent cystoscopy between November 1999 and February 2002 to evaluate febrile UTIs in 40 patients, dysfunctional voiding in 14 and hydronephrosis in 3. The control group (27 patients, 54 renal units) was used to assess the accuracy of PIC by comparing the results against those with the standard cystogram in children who did not have febrile infection and did not demonstrate vesicoureteral reflux (15 patients, 30 renal units), and in those who had febrile infection and vesicoureteral reflux (12 patients, 24 renal units). The study group (30 patients, 60 renal units) served to assess the incidence of "occult" vesicoureteral reflux in children who experienced febrile UTIs yet did not have vesicoureteral reflux on standard cystography. RESULTS: CONTROL GROUP: In children without febrile UTIs all 30 ureteral orifices had a normal endoscopic appearance and no vesicoureteral reflux by PIC cystography. In children with febrile UTIs 15 ureteral orifices with known vesicoureteral reflux were lateral and/or patulous and demonstrated vesicoureteral reflux by PIC, 4 appeared normal and did not exhibit vesicoureteral reflux on standard cystography or by PIC, and 5 were lateral and/or patulous in appearance and did not display vesicoureteral reflux on standard cystography, but did show vesicoureteral reflux by PIC. These findings reveal that PIC cystography is 100% sensitive at demonstrating reflux already known by standard cystogram, is 87% specific as it showed reflux in 5 of 39 renal units not revealed by a standard cystogram and has an overall accuracy of 91%. STUDY GROUP: At cystoscopy all 30 children demonstrated an abnormal appearance of one or both ureteral orifices. PIC cystography showed vesicoureteral reflux in all 30 children (48 renal units, 12 unilateral and 18 bilateral). The remaining ureteral orifices (12), which appeared normal, did not permit vesicoureteral reflux. Children with vesicoureteral reflux by PIC were treated with antimicrobial prophylaxis (26) or ureteral reimplantation (4, 2 unilateral and 2 bilateral reimplantation) Postoperatively, these children did not experience a febrile UTI during followup (average 8 months). CONCLUSIONS: PIC cystography is simple to perform using routinely available operating room equipment and does not artifactually induce vesicoureteral reflux. The incidence of "occult" vesicoureteral reflux in children who experience febrile UTIs without vesicoureteral reflux on standard cystography is 100% by PIC cystography. PIC cystography should be done when vesicoureteral reflux is suspected in children who experience febrile UTIs but do not exhibit reflux on standard cystography. PMID- 12771796 TI - Interobserver and intra-observer agreement in interpreting urodynamic measurements in children. AB - PURPOSE: We evaluated the comparability and repeatability of analyses based on urodynamic reports. MATERIALS AND METHODS: We assessed interobserver and intra observer agreement when interpreting urodynamic examinations performed in children. Four pediatric urologists analyzed 17 sets of urodynamic data and gave their interpretations of the possible underlying condition of the tested subjects. The consensus among observers was analyzed using the kappa statistic. RESULTS: Poor agreement was observed on suggested diagnoses based on uroflow curves. Good consensus was noted on maximal flow rates but the interpretations of urethral pressure profile curves and the resulting diagnoses regarding sphincter function differed significantly. It also appeared to be difficult to reach agreement regarding bladder compliance and detrusor activity. CONCLUSIONS: The consistency of the interpretations of urodynamic studies should be improved before they are accepted as measures for comparing urodynamic diagnoses or effects of treatment between patient series. PMID- 12771798 TI - Laparoscopic ureterocalicostomy: a feasibility study. AB - PURPOSE: Ureterocalicostomy is occasionally indicated for reconstruction of recurrent, recalcitrant ureteropelvic junction obstruction associated with postoperative fibrosis and a relatively inaccessible renal pelvis. We investigated the feasibility of performing laparoscopic ureterocalicostomy in a survival porcine model. Anatomical, histological and chronic functional outcomes were evaluated. MATERIALS AND METHODS: Laparoscopic ureterocalicostomy was performed in 10 survival female swine. A ureteropelvic junction obstruction model was created by laparoscopic ligation of a 2 to 3 cm. segment of upper ureter. After an interval of complete ureteropelvic junction obstruction laparoscopic ureterocalicostomy was performed in a manner duplicating the steps of conventional open surgery. After transverse amputation of the lower renal pole end-to-end anastomosis of the proximal ureter to the inferior calix was formed by laparoscopic freehand suturing and knot-tying techniques. RESULTS: Mean ureter stricture length was 2.2 cm. (range 1.7 to 3.1). Mean duration of obstruction before laparoscopic ureterocalicostomy was 6.3 days (range 2 to 18). Mean operative time for laparoscopic ureterocalicostomy was 165.3 minutes (range 105 to 240). Mean estimated blood loss was 145 cc (range 25 to 400). Mean stent duration in 6 pigs was 8.7 days (range 7 to 11). Excretory urograms demonstrated immediate function with symmetrical and unobstructed drainage in all operated renal units. At 4 to 8 weeks of followup no urine leaks were noted and histological examination documented complete urothelial healing without fibrosis or scar formation. CONCLUSIONS: Laparoscopic ureterocalicostomy is technically feasible in the porcine model and it effectively duplicates the established principles of open surgery. Our technique further extends the application of laparoscopic surgery for difficult ureteropelvic junction obstruction. PMID- 12771797 TI - Cyclooxygenase-2 as a potential target in the prevention and treatment of genitourinary tumors: a review. AB - PURPOSE: Recent years have seen a dramatic expansion in our discovery and knowledge of the molecular mechanisms of cancer development and progression. The discovery and elucidation of prostaglandin pathways, in particular the molecular and clinical role of cyclooxygenase (COX)-2 function, has had important application to neoplasms. Current understanding of the role of COX-2 activity and, thereby, the potential clinical usefulness of COX-2 specific inhibitors as they apply to urological oncology are discussed. MATERIALS AND METHODS: The discovery of prostaglandin pathways, the molecular and clinical role of COX-2 function, and the corresponding application to neoplasms were reviewed in the scientific literature (MEDLINE from 1960 to the present). In particular, a thorough review of the current literature and recent abstract presentations at scientific meetings was done regarding the potential role of COX-2 in urological cancers (MEDLINE from 1960 to the present, and American Urological Association and American Society of Clinical Oncology annual meeting abstracts from1998 to the present). RESULTS: Decreased apoptosis, increased angiogenesis and immunosuppression are just some of the known sequelae of COX-2 over expression and each effect may have an important role in tumor formation and progression. Preclinical research and pilot clinical studies in urological oncology, in particular prostate, bladder and kidney cancer, have proved to be quite promising to date. CONCLUSIONS: Currently we are just beginning to understand the molecular mechanisms and clinical effects of COX-2 function and inhibition, and the potential for COX-2 specific inhibitors to affect potentially tumor biology and growth and, thereby, serve as antitumor drugs with therapeutic and chemopreventive roles for urological cancers. The absence of complete scientific understanding in these areas provides a generous opportunity for innovative and important scientific study. PMID- 12771799 TI - DNA mismatch repair genes in renal cell carcinoma. AB - PURPOSE: DNA mismatch repair is one of the correcting mechanisms that preserves genetic stability during replication or chemically induced damage. We hypothesized that genetic instability is due to a defect in mismatch repair genes in renal cell carcinoma. To test this hypothesis mismatch repair genes hMLH1, hMSH2, hMSH3, hMSH6, hPMS1 and hPMS2 were analyzed in renal cell carcinoma cell lines and tissues. We further investigated the mechanisms of inactivation of these genes through CpG methylation pathways. MATERIALS AND METHODS: We analyzed 41 fresh normal and renal cell carcinoma samples for gene and protein expression of various mismatch repair genes (hMLH1, hMSH2, hMSH3, hMSH6, hPMS1 and hPMS2) using reverse transcriptase-polymerase chain reaction and immunohistochemistry techniques. To investigate the mechanisms of inactivation of these genes cultured renal cancer cell lines (A498, Caki-1 and Caki-2) were treated with demethylating agent (5-aza-2'-deoxycytidine), and mismatch repair genes and protein expression were analyzed before and after treatment. RESULTS: hMLH1 and hMSH3 mRNA expression was significantly lower in renal cell carcinoma tissues than in normal tissues. Similarly nuclear positivity of hMSH3 was significantly lower in renal cell carcinoma tissues than in normal tissues. Moreover, at the mRNA and protein levels hMSH3 expression in high grade renal cell carcinomas was significantly lower than in low grade tumors. However, there was no significant difference in hMSH2, hMSH6, hPMS1 or hPMS2 expression in renal cell carcinoma tissues versus normal kidney tissues. In renal cancer cell lines demethylation with 5-aza-2' deoxycytidine did not affect the expression of hMLH1 and hMSH3 genes. CONCLUSIONS: To our knowledge this is the first comprehensive study demonstrating the down-regulation of mismatch repair genes in renal cell carcinoma. Selective defect in some mismatch repair genes can cause genomic instability and activate the malignant transformation as well as the progression of renal cell carcinoma. PMID- 12771800 TI - Induction of apoptosis in human bladder cancer cells in vitro and in vivo caused by FTY720 treatment. AB - PURPOSE: FTY720 is a unique immunosuppressant that induces apoptosis in activated lymphocytes but not in other hematopoietic cells. We examined whether FTY720 has anticancer effects on human bladder cancer cells by inducing apoptosis and we investigated its molecular pathway. MATERIALS AND METHODS: We used the 3 human bladder cancer cell lines T24, UMUC3 and HT1197, and the human fibroblast derived cell line CRL-2096 (American Type Tissue Collection, Rockville, Maryland) in this study. The difference in drug susceptibility to FTY720 in cancer cells and fibroblasts was examined by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide) and cell growth assays. FTY720 induced apoptosis was determined by morphological analysis under light and electron microscopy, and DNA electrophoresis, and its molecular pathway was evaluated by Western blot analysis focusing on the p42/p44 mitogen activated protein kinase pathway. We then tested the in vivo effect of this agent using 2 mouse models of human bladder cancer xenograft. RESULTS: FTY720 treatment in vitro induced selective apoptosis in cancer cells at a concentration of less than 10 microM. Morphological analysis revealed features characteristic of apoptosis, including small cytoplasm with fragmented nuclei and condensed chromatin. DNA electrophoresis confirmed apoptosis, as evidenced by a distinct oligosomal ladder. Western blot analysis revealed that the agent significantly inhibited hepatocyte growth factor induced p42/p44 mitogen activated protein kinase activity. The in vivo anticancer effect was clearly confirmed by significantly decreased tumor growth without notable side effects in the 2 xenograft models. CONCLUSIONS: FTY720 treatment may induce selective apoptosis in vitro as well as in vivo in cancer cells. We suggest that FTY720 is a potent and clinically applicable anticancer agent for bladder cancer. PMID- 12771801 TI - Association of V89L SRD5A2 polymorphism with prostate cancer development in a Japanese population. AB - PURPOSE: The SRD5A2 gene codes the steroid 5-reductase type II, a critical mediator of androgen action, and the V89L and A49T polymorphisms of this gene may be associated with a distinct enzyme activity. We explored the association among these polymorphisms and the risk of prostate cancer or benign prostatic hyperplasia (BPH) in a Japanese population. MATERIALS AND METHODS: This study included 302 patients with prostate cancer, 228 with BPH and 243 male controls. V89L and A49T polymorphisms were analyzed by the polymerase chain reaction restriction fragment length polymorphism method. Genotypes were evaluated by electrophoresis on agarose gel. RESULTS: For the V89L polymorphism there were no significant differences in genotype frequencies in patients with prostate cancer and controls (p = 0.071) or in patients with BPH and male controls (p = 0.219). However, males with the VV or VL genotype were at significantly increased risk for prostate cancer compared with those with the LL genotype (adjusted OR 1.69, 95% CI 1.07 to 2.65, p = 0.024). The risk of BPH in males with the VV or VL genotype was not significantly elevated in comparison with those with the LL genotype (adjusted OR 1.37, 95% CI 0.85 to 2.20, p = 0.194). The V89L variant was not associated with the grade or stage of prostate cancer, or with patient age. For the A49T polymorphism all subjects had the AA genotype. CONCLUSIONS: The V allele of the V89L polymorphism in the SRD5A2 gene may dominantly increase the risk of prostate cancer. PMID- 12771802 TI - Characterization of Escherichia coli strains causing urinary tract infections in patients with transposed intestinal segments. AB - PURPOSE: Transposition of intestinal segments into the urinary tract predisposes to urinary tract infections. We characterized bacterial infections in these patients and examined the virulence genotype and persistence of Escherichia coli isolates. MATERIALS AND METHODS: We followed 26 patients who underwent bladder reconstructive surgery using transposed intestinal segments. E. coli strains isolated from the urine of these patients were genotyped for established virulence determinants and the frequency of carriage was compared with E. coli strains isolated from community acquired urinary infections and the fecal flora of anonymous volunteers. A longitudinal study of E. coli strains in 9 patients was also done using pulsed field gel electrophoresis. RESULTS: E. coli was the most frequently isolated organism, responsible for 59% (62 of 105) of monobacterial infections. Other bacteria isolated included Klebsiella species, Proteus species and Enterococcus faecalis. Community acquired E. coli strains were more likely to carry multiple determinants for particular adhesins (P and S fimbriae) and toxins (alpha-hemolysin and cytotoxic necrotizing factor) than fecal strains. Carriage frequency for bladder reconstruction strains was intermediary and not significantly different. The key finding was that E. coli strains persisted for prolonged periods, including 2 years in certain patients, often despite various antimicrobial treatments. CONCLUSIONS: This study highlights that further steps must be taken to prevent and treat urinary tract infections in this susceptible group. Particular attention should be given to the treatment of persistent infections. PMID- 12771803 TI - Signaling through PI3K/Akt mediates stretch and PDGF-BB-dependent DNA synthesis in bladder smooth muscle cells. AB - PURPOSE: Smooth muscle cells (SMC) of the bladder undergo hypertrophy and hyperplasia following exposure to sustained mechanical overload. Although superficial similarities in the response of the heart and bladder to hypertrophic stimuli suggest that similar molecular mechanisms may be involved, this remains to be demonstrated. In this study we compared signal transduction pathway activation in primary culture bladder SMC and cardiac myofibroblasts in response to cyclic stretch. The effects of growth factor stimulation on pathway activation in bladder SMC were also investigated. MATERIALS AND METHODS: Primary culture rodent bladder SMC or cardiac myofibroblasts were subjected to cyclic stretch relaxation in the absence or presence of pharmacologic inhibitors of the phosphoinositide-3-kinase, (PI3K)/Akt, extracellular signal-regulated kinase mitogen activated protein kinase (Erk-MAPK) or the p38 stress-activated protein kinase-2 (SAPK2) pathways. In parallel experiments human bladder SMC were treated with platelet-derived growth factor-BB (PDGF-BB), heparin-binding EGF-like growth factor (HB-EGF) or fibroblast growth factor-2 (FGF-2). In each case the extent of DNA synthesis was determined by uptake of tritiated thymidine, and activation of specific signaling intermediates was determined by immunoblot analysis using antibodies to the non-phosphorylated and phosphorylated (activated) forms of Akt, p38 and Erk1/2. RESULTS: Akt and p38 were rapidly phosphorylated in stretched bladder SMC and cardiac myofibroblasts, and stretch-induced DNA synthesis in these cells was ablated with inhibitors of PI3K or p38 but not Erk-MAPK. Similarly, PDGF-BB up-regulated DNA synthesis in bladder SMC in a p38 and Akt dependent manner. CONCLUSIONS: We conclude that distinct stimuli, such as mechanical stretch and PDGF-BB, promote DNA synthesis in bladder SMC through shared downstream signaling pathways. Furthermore, phenotypically similar cells from the bladder and heart show comparable pathway activation in response to stretch. These findings suggest that similar molecular mechanisms underlie the altered growth responses of the bladder and heart to mechanical overload. This study also provides the first report of Akt activation in bladder SMC and suggests that Akt, consistent with its pivotal role in cardiac hypertrophy, may also be a key regulator of remodeling in the SMC compartment of the bladder exposed to hypertrophic/hyperplastic stimuli in vivo. PMID- 12771804 TI - Phosphoinositide signaling in urological disease. PMID- 12771805 TI - Functional response of bladder strips from streptozotocin diabetic rats depends on bladder mass. AB - PURPOSE: We investigated the relationship of bladder mass to responses to electrical field stimulation and adrenergic agonists in diabetic rat bladders. MATERIALS AND METHODS: Longitudinal strips were removed from the ventral and dorsal detrusor of age matched control, 2-month diabetic and sucrose drinking rats. Contractile responses to electrical field stimulation, KCl and phenylephrine, and relaxation in response to norepinephrine and isoproterenol were measured. RESULTS: Bladders from sucrose drinking and diabetic rats weighed significantly more than those of controls. Diabetic rats were divided into 2 groups with the bladder weighing less than or greater than 265 mg. Strips from small diabetic bladders were generally more responsive to field stimulation and norepinephrine than those from control or sucrose drinking rats. Conversely decreased function was especially apparent in dorsal strips from large diabetic bladders. Ventral strips were significantly more sensitive to the relaxant actions of norepinephrine and isoproterenol than dorsal strips. CONCLUSIONS: Our results suggest that the responsiveness of diabetic rat bladder to electrical field stimulation and adrenergic agonists is related to bladder mass, analogous to observations after partial outlet obstruction. Decreased function was particularly apparent in dorsal strips from diabetic rats with a large bladder. PMID- 12771806 TI - Acute intravesical infusion of a cobalt solution stimulates a hypoxia response, growth and angiogenesis in the rat bladder. AB - PURPOSE: Experimental partial bladder outlet obstruction of rats induces a bladder growth and remodeling process similar to that in humans with benign prostatic hyperplasia. Previously we have proposed that bladder hypoxia associated with partial bladder outlet obstruction is a stimulus of this bladder growth process. We report our results of testing the acute effects of a simple chemical agent (cobaltous ion) known to mimic hypoxia in the rat bladder. We measured its ability to effect bladder gene expression, angiogenesis and growth processes. MATERIALS AND METHODS: Adult rats were divided into 2 groups. One group (controls) received intravesical saline 3 times for 30 minutes in 6 days and the other received intravesical saline with 100 microM. CoCl(2) at the same times. All animals also received continuous infusion of BrdU for the 6-day period through an implanted osmotic pump. Portions of the bladders from these rats were fixed, sectioned, stained for microscopic analysis and immunohistochemically stained to identify BrdU positive cells and vascular elements via factor VIII staining. Other portions were frozen, extracted for proteins and the proteins were comparatively analyzed for the expression of hypoxia inducible factor-1alpha and vascular endothelial growth factor on Western blots. RESULTS: Bladders infused with CoCl(2) showed extensive expansion of the submucosal region, which was significant compared with that in saline infused bladders. Cells in this expanded region as well as cells within the urothelium were found to be extensively labeled with BrdU, in contrast to control bladders, which had rare BrdU labeled cells in any region. Immunohistochemical analysis for factor VIII showed that the submucosal region of cobalt treated rats contained numerous small vessels and microvessels that were not apparent in controls. These cellular changes were consistent with our finding of increased hypoxia inducible factor 1alpha and vascular endothelial growth factor protein expression in cobalt treated bladders compared with controls. CONCLUSIONS: Acute intravesical instillation of cobalt ion solution into the rat bladder initiated a hypoxia response accompanied by increased bladder angiogenesis and growth. This finding supports the idea that hypoxia is a stimulus for bladder growth subsequent to partial bladder outlet obstruction. PMID- 12771807 TI - Expression of nitric oxide synthase immunoreactivity in the human female intramural striated urethral sphincter. AB - PURPOSE: While we have recently detected neuronal nitric oxide synthase (nNOS) immunoreactivity in a heterogeneous population of human male urethral striated muscle, to our knowledge the association of nNOS in the female counterpart is unknown. We investigated the association of nNOS with female urethral striated muscle and re-investigated muscle fiber types. MATERIALS AND METHODS: Cryostat sections were taken from the middle third of 4 human female urethras. Nicotinamide adenine dinucleotide phosphate diaphorase histochemistry and nNOS immunohistochemistry were performed. Muscle fiber types were identified by myofibrillar adenosine triphosphatase histochemistry and fast twitch troponin T immunohistochemistry. The association between nNOS immunoreactivity and muscle fiber type was analyzed. RESULTS: Positive staining for nicotinamide adenine dinucleotide phosphate diaphorase histochemistry and nNOS immunoreactivity were recognized in the sarcolemma of 43.9% of female urethral striated muscle fibers. Immunoreactivity for fast twitch troponin T immunohistochemistry was demonstrated by 2% of the striated fibers. The use of myofibrillar adenosine triphosphatase showed that all fibers darkly stained uniformly at a pH of 9.6, 4.6 and 4.3, suggesting that they were myofibrillar intermediate muscle fibers. The results allowed the differentiation of 2 subgroups of fibers, namely smaller fibers (modal diameter 10.1 to 15.0 microm.) without nNOS immunoreactivity and larger fibers (modal diameter 15.1 to 20.0 microm.) with nNOS immunoreactivity. CONCLUSIONS: To our knowledge female urethral striated muscle has for the first time been found to consist of myofibrillar intermediate fibers and nNOS was positively localized in the sarcolemma of a subgroup of the fibers. This study provides a basis for further investigation into female urethral striated sphincter function and changes in pathological conditions. PMID- 12771810 TI - Knee kinematics and total knee replacement design. PMID- 12771808 TI - Modulation of Ca2+ sensitivity regulates contractility of rabbit corpus cavernosum smooth muscle. AB - PURPOSE: We examined the role of the modulation of Ca2+ sensitivity for regulating the contractility of corpus cavernosum smooth muscle. MATERIALS AND METHODS: We applied simultaneous measurements of intracellular Ca2+ concentration and tension in fura-PE3 loaded intact strips and receptor coupled permeabilization by alpha-toxin. RESULTS: In intact fura-PE3 loaded strips the tension induced by 10 microM. phenylephrine was significantly greater than that produced by depolarization with 118 mM. K+, although the extent of intracellular Ca2+ concentration elevations was similar. During sustained contraction induced by 10 microM. phenylephrine the application of 10 microM. Y-27632 (a Rho kinase inhibitor) induced relaxation with a slight decrease in intracellular Ca2+ concentration, while the application of 3 microM. GF109203X (a protein kinase C inhibitor) induced relaxation without changing intracellular Ca2+ concentration. In alpha-toxin permeabilized strips 10 microM. phenylephrine induced a larger increase in force at a constant intracellular Ca2+ concentration and produced a leftward shift in the intracellular Ca2+ concentration-tension relationship, a response that was partially inhibited by pretreatment with Y-27632 or GF109203X. CONCLUSIONS: These results indicate that in rabbit corpus cavernosum smooth muscle phenylephrine induces contraction not only by increasing intracellular Ca2+ concentration, but also by increasing Ca2+ sensitivity of the contractile apparatus in a Rho kinase and protein kinase C dependent manner. Antagonism of Ca2+ sensitization pathways in the corpus cavernosum smooth muscle represents an alternate target for the treatment of erectile dysfunction. PMID- 12771812 TI - The anatomic literature relating to the knee from 1836 to 1917: an historic note. PMID- 12771811 TI - Joint laxity as a criterion for the design of condylar knee prostheses. 1974. PMID- 12771813 TI - Functional morphologic features of the human knee: an evolutionary perspective. AB - The complex functional morphologic characteristics of the knee are of ancient origin. The multiple asymmetries of anatomy can be traced back more than 300 million years to the pelvic appendages of Sarcoptorigian lobe-finned fish. The knee functions as a biologic transmission with ligaments acting as sensate linkages and the menisci acting as sensate, mobile bearings. Cine-computed tomography and magnetic resonance imaging studies of intact knees from cadavers reveal a combined rolling and gliding motion, with posterior displacement of the femorotibial contact point with increasing flexion. The posterior displacement of the femorotibial contact point is greater in the lateral compartment by approximately a factor of two. The anatomy of the lateral compartment, including the inferior sloping of the posterior tibial plateau, reflects and accommodates this greater motion. This asymmetry of kinematics between the medial and lateral compartment, an established characteristic of human and many other extant mammalian knees, results in an internal rotation of the tibia relative to femur with increasing flexion. By taking into account the complexities of functional morphologic features of the knee, the design of joint replacements and bracing systems may be improved. PMID- 12771814 TI - Kinematics of the human knee using an open chain cadaver model. AB - There continues to be controversy about the kinematics of the human knee. This study used seven knees from cadavers moved by pulling on the quadriceps tendon in an open chain fashion using video motion analysis to determine the instantaneous helical axis of movement. Computed tomography scans of the specimens allowed the axes to be related to condyles. The parameter beta was defined by the relationship of the helical axis to the center of the condyle (pure spinning motion) and the contact point of the condyle on the tibia (pure rolling motion). Axes above the center of the condyle represent countertranslation, those between the center and the contact point combined spinning and rolling, and those below represent concordant translation. If the motion of the knee is guided by the crossed four-bar link then this model, that allows the knee to 'seek its own path' throughout the range of motion, should show the rollback that commonly is thought to be an important feature of knee motion. The results of this study show that the medial side of the knee stays stable in spinning kinematics whereas the lateral side has a rolling motion in full flexion progressing to a spinning motion in midflexion and counter-translation near full extension. The kinematics that would be expected from rollback were not observed. PMID- 12771815 TI - The movement of the knee studied by magnetic resonance imaging. AB - The author's work using magnetic resonance imaging to study the relative movements (the kinematics) of the tibia and femur is reviewed. The description is understood best by reference to comparative anatomy and by dividing the flexion arc into three components. Knee activities take place mainly between 10 degrees and 120 degrees. Over this arc, the articulating surfaces of the femoral condyles are circular in sagittal section and rotate around their center. The medial condyle does not move anteroposteriorly (roll-back does not occur medially). The lateral condyle tends to roll back producing tibial internal rotation with flexion. From full extension to 10 degrees to 30 degrees tibial internal rotation is coupled with flexion. The articulating surfaces medially are a larger radiused anterior femoral facet, which articulates with an upward-sloping tibial facet. Laterally, the femoral condyle rolls forward onto the anterior horn. Flexion beyond 120 degrees only can be achieved passively. Medially, the femur rolls up onto the posterior horn. Laterally, the femur and the posterior horn drop over the posterior tibia. PMID- 12771816 TI - The use of functional analysis in evaluating knee kinematics. AB - The importance of understanding the six-degrees-of-freedom kinematics of the knee during ambulatory activities was examined in the context of the function of total knee arthroplasty. Studies of knee kinematics during walking, stair climbing, and a deep flexion squat indicate that knee kinematics is activity-dependent. A comparative study of patients and healthy subjects during stair climbing indicates the importance of maintaining the function of the posterior cruciate ligament. A second study used walking kinematics derived from patient testing as input to a wear simulator. There was increased wear relative to standard simulator input that was related to the slip velocity at the contact surface. Finally, results from a study of deep flexion indicate that substantial femoral rotation is required during deep flexion activities. The current study shows the importance of studying in vivo knee kinematics for future enhancement in the treatment of the arthritic knee. PMID- 12771817 TI - Characterizing the motion of total knee replacements in laboratory tests. AB - The purposes of determining the motion of a total knee replacement in vitro are to characterize the stability and laxity characteristics and to predict the kinematic behavior of the total knee replacement when implanted. With this information, different total knee replacement designs can be compared, the stability of the implanted knee replacement with and without the retention of ligaments can be assessed, and tests can be formulated to measure the wear and deformation of the materials, the strength of the components, and the fixation to the bone. Initially, different ways are described for characterizing the kinematics of the natural knee. We propose a rationale for the kinematic testing of total knee replacements in vitro using mechanical tests. One of the key questions is whether there is an objective method of measuring the kinematics of a total knee replacement in vitro, which will relate to or predict its behavior in vivo. PMID- 12771818 TI - In vivo fluoroscopic analysis of the normal human knee. AB - The objective of the current study was to use fluoroscopy and computed tomography to accurately determine the three-dimensional, in vivo, weightbearing kinematics of five normal knees. Three-dimensional computer-aided design models of each subject's femur and tibia were recreated from the three-dimensional computed tomography bone density data. Three-dimensional motions for each subject then were determined for five weightbearing activities. During gait, the lateral condyle experienced -4.3 mm (range, -1.9--10.3 mm) of average motion, whereas the medial condyle moved only -0.9 mm (range, 3.4--5.8 mm). One subject experienced 5.8 mm of medial condyle motion. On average, during deep flexion activities, subjects experienced -12.7 mm (range, 1.4--29.8 mm) of lateral condyle motion, whereas the medial condyle motion only was -2.9 mm (range, 3.0--9.0 mm). One subject experienced 5.8 and 9.0 mm of medial condyle motion during gait and a deep knee bend, respectively leading to the occurrence of a lateral pivot motion. During the deep flexion activities, the subjects experienced significantly more axial rotation (> 13 degrees) than gait (< 5 degrees). During all five activities, the lateral condyle experienced significantly more anteroposterior translation, leading to axial rotation of the tibia relative to the femur. PMID- 12771819 TI - Rotational constraint in posterior-stabilized total knee prostheses. AB - Rotational stresses from box-post impingement have been implicated in the loosening of posterior-stabilized total knee prostheses. A bench model was constructed to assess the forces generated by tibiofemoral rotation. Rotational torque under load was measured in two different posteriorstabilized total knee prostheses using an axial-torsion load cell at 0 degrees, 20 degrees, and 40 degrees flexion over 20 degrees internal and external rotation. The Sigma posterior-stabilized prosthesis generated little torque through 5 degrees internal and external rotation. An increase in torque then occurred because of box-post impingement, generating peak torques of 17 to 18 N-m at 12 degrees to 14 degrees rotation. The bench model produced the same deformation of the polyethylene post as seen on retrieved specimens. The Scorpio posterior stabilized prosthesis had a relatively continuous rise in generated torque from tibiofemoral conformity. Box-post impingement did not occur resulting in 32% lower torque between 12 degrees and 14 degrees rotation. Peak rotational torques of 15 to 16 N-m were reached at 19 degrees to 20 degrees rotation. Tibiofemoral conformity is the primary source of rotational constraint. Box-post impingement can be a source of additional rotational constraint. Depending on specific design features, small changes in relative tibiofemoral component rotation can more than double the generated torque. Axial rotation of the knee in vivo can generate substantial torque. Relative tibiofemoral rotational position is an important factor influencing component function and fixation. PMID- 12771820 TI - The effects of external torque on polyethylene tibial insert damage patterns. AB - The forces and torques that occur during walking gait, particularly during toe off, promote articulation in the posteromedial quadrant of tibial inserts. Retrieved components of failed knee arthroplasties show ultrahigh molecular weight polyethylene damage patterns in this region. Component-designed constraint, compromised polymer, and surgical factors account for these observations. The current authors compare the contact stresses that developed on four implant designs during toe-off for optimally aligned and externally torqued components using the finite element method. Under 16 N-m of torque, the four designs studied varied regarding their centers of rotation and magnitude of external rotation, which are related directly to their specific articulating surface geometry. Designs with conforming condylar geometry had greater rotational constraint and therefore, less external rotation. These conforming designs offer the benefits of lower stresses and tend to limit contact near the edge of the plateau. However, because of their increased rotational constraint, torque is transmitted more readily to the implant-bone interface, increasing the potential for implant loosening. The data presented serve as an indicator of the potential for polyethylene tibial component surface damage and define the role that implant geometry plays in resisting external rotation. PMID- 12771821 TI - Femoral rollback after cruciate-retaining and stabilizing total knee arthroplasty. AB - Limited data comparing the kinematics of posterior cruciate ligament-retaining or substituting total knee arthroplasty with its own intact knee under identical loadings is available. In the current study, posterior femoral translation of the lateral and medial femoral condyles under unloaded conditions was examined for intact, cruciate-retaining, cruciate ligament-deficient cruciate-retaining and posterior-substituting knee arthroplasties. Cruciate-retaining and substituting total knee arthroplasties behaved similarly to the cruciate-deficient cruciate retaining total knee arthroplasty between 0 degrees and 30 degrees flexion. Beyond 30 degrees, the posterior cruciate-retaining arthroplasty showed a significant increase in posterior translation of both femoral condyles. The posterior cruciate-substituting arthroplasty only showed a significant increase in posterior femoral translation after 90 degrees. At 120 degrees, both arthroplasties restored approximately 80% of that of the native knee. Posterior translation of the lateral femoral condyle was greater than that observed in the medial condyle for all knees, indicating the presence of internal tibial rotation during knee flexion. The data showed that the posterior cruciate ligament is an important structure in posterior cruciate-retaining total knee arthroplasty and proper balancing is imperative to the success of the implant. The cam-spine engagement is valuable in restoring posterior femoral translation in posterior cruciate-substituting total knee arthroplasty. PMID- 12771822 TI - In vivo fluoroscopic analysis of fixed-bearing total knee replacements. AB - In vivo kinematic patterns were determined for subjects (patients participating in the study), having either a fixed-bearing posterior-stabilized or posterior cruciate-retaining total knee arthroplasty. While under fluoroscopic surveillance, subjects did normal gait and a deep knee bend. Video images were downloaded to a workstation computer and analyzed in three dimensions using an iterative model-fitting approach. Femorotibial contact paths for the medial and lateral condyles, axial rotation, and condylar lift-off were determined. During a deep knee bend, subjects having a posterior-stabilized total knee arthroplasty routinely experienced posterior femoral rollback of their lateral condyle and normal axial rotational patterns, whereas random subjects having a posterior cruciate-retaining total knee arthroplasty experienced paradoxical anterior sliding and opposite axial rotational patterns. During gait, posterior-stabilized and posterior cruciate-retaining total knee arthroplasties experienced similar kinematic patterns, with the presence of paradoxical sliding and opposite axial rotational patterns. Subjects having posterior-stabilized and posterior cruciate retaining total knee arthroplasties experienced condylar lift-off. Subjects having a posterior cruciate-retaining total knee arthroplasty predominantly experienced lateral condylar lift-off whereas subjects with posterior-stabilized total knee arthroplasties experienced either medial or lateral condylar lift-off. Subjects having a posterior-stabilized total knee arthroplasty experienced significantly greater weightbearing range of motion. PMID- 12771823 TI - Knee motions during maximum flexion in fixed and mobile-bearing arthroplasties. AB - Full flexion is a critical performance requirement for patients in Asia and the Middle East, and increasingly for patients in Europe and North America who have total knee arthroplasty. There has been considerable work characterizing maximum flexion in terms of clinical, surgical, and preoperative factors, but less in vivo experimental work after rehabilitation. The purpose of the current investigation was to determine whether anteroposterior tibiofemoral translation influenced maximum weightbearing knee flexion in patients with good or excellent clinical and functional outcomes. One hundred twenty-one knees in 93 subjects, including 16 different articular surface designs, were studied using fluoroscopy and shape matching to determine knee kinematics in a weightbearing deep flexion activity. A relatively posterior position of the femur on the tibia was significantly correlated with greater maximum knee flexion. Posterior-stabilized arthroplasties had significantly more posterior femoral position and maximum flexion than posterior cruciate-retaining fixed-bearing arthroplasties, which had more posterior femoral position and greater maximum flexion than mobile-bearing arthroplasties. Posterior bone-implant impingement was observed in 28% of knees. Tibiofemoral motions influence the mechanics of weightbearing deep flexion in well-functioning knee arthroplasties. PMID- 12771824 TI - Fluoroscopic analyses of cruciate-retaining and medial pivot knee implants. AB - Contemporary posterior cruciate-retaining total knee designs have provided pain relief and improved knee function but have failed to reproduce the kinematics and stability of the normal nonarthritic knee. The Medial Pivot total knee design features a near constant radius of curvature of the femoral component. The tibial surface is highly congruent and asymmetric, permitting a medial pivot motion during knee flexion. The purpose of the current study was to analyze and compare the gait kinematics of the Sigma posterior cruciate-retaining total knee implant, the Advance Traditional posterior cruciate-retaining total knee implant, and the Advance Medial Pivot knee implant using fluoroscopic analysis. In vivo kinematics were determined for 15 clinically successful total knee arthroplasties. Five knee implants were evaluated from each group. The authors analyzed the kinematics of knee motion during the stance phase of gait for each patient. On average, subjects with the Medial Pivot knee implant had a medial pivot motion. Both posterior cruciateretaining designs had a paradoxical roll forward of the tibia on femur during knee flexion and had greater excursion of both condyles during knee flexion than the medial pivot design. Nine of 10 of the posterior cruciate retaining designs had condylar lift-off averaging 1.7 mm whereas only one Medial Pivot knee implant had condylar lift-off measuring 1.1 mm. PMID- 12771825 TI - The impact of femoral component rotational alignment on condylar lift-off. AB - The surgical technique and implant design influence femoral condylar lift-off in total knee arthroplasty. With the use of the classic method of bone resection and appropriate soft tissue releases, alignment of the femoral component along the transepicondylar axis will create a symmetric and balanced flexion space. This surgical technique will reduce the incidence of condylar lift-off. If condylar lift-off does occur, it may be beneficial to have a prosthesis that is designed with a conforming frontal articulation. Reducing lift-off with a relatively conforming articulation reduces edge loading and in turn should reduce polyethylene damage. PMID- 12771826 TI - Metrology to quantify wear and creep of polyethylene tibial knee inserts. AB - Assessment of damage on articular surfaces of ultrahigh molecular weight polyethylene tibial knee inserts primarily has been limited to qualitative methods, such as visual observation and classification of features such as pitting, delamination, and subsurface cracking. Semiquantitative methods also have been proposed to determine the linear penetration and volume of the scar that forms on articular surfaces of tibial knee inserts. The current authors report a new metrologic method that uses a coordinate measuring machine to quantify the dimensions of this scar. The articular surface of the insert is digitized with the coordinate measuring machine before and after regular intervals of testing on a knee simulator. The volume and linear penetration of the scar are calculated by mathematically taking the difference between the digitized surface maps of the worn and unworn articular surfaces. Three conventional polyethylene tibial knee inserts of a posterior cruciate-sparing design were subjected to five million cycles of normal gait on a displacement driven knee wear simulator in bovine serum. A metrologic method was used to calculate creep and wear contributions to the scar formation on each tibial plateau. Weight loss of the inserts was determined gravimetrically with the appropriate correction for fluid absorption. The total average wear volume was 43 +/- 9 and 41 +/- 4 mm3 measured by the metrologic and gravimetric methods, respectively. The wear rate averaged 8.3 +/- 0.9 and 8.5 +/- 1.6 mm3 per million cycles measured by the metrologic and gravimetric methods, respectively. These comparisons reflected strong agreement between the metrologic and gravimetric methods. PMID- 12771827 TI - Polyethylene wear particles in synovial fluid after total knee arthroplasty. AB - The aims of the current study were to examine polyethylene particles in synovial fluid at an early stage, and to compare a newly introduced medial pivot total knee prosthesis with an established posterior-stabilized total knee prosthesis. Synovial fluid was obtained 1 year after knee arthroplasty from 17 patients with well-functioning prostheses (22 knees, 11 posterior-stabilized prostheses and 11 medial pivot prostheses) under complete sterile conditions. Polyethylene particles were isolated and analyzed by scanning electron microscopy. Particle size (equivalent circle diameter) was 0.78 +/- 0.08 microm (mean +/- standard error) in posterior-stabilized prostheses and 0.67 +/- 0.06 microm in medial pivot prostheses. Particle shape (aspect ratio) was 2.30 +/- 0.22 in posterior stabilized prostheses and 1.90 +/- 0.16 in medial pivot prostheses. The total numbers of particles were 1.16 +/- 0.57 x 10(8) in posterior-stabilized prostheses and 9.01 +/- 2.95 x 10(6) in medial pivot prostheses. Particles were smaller and rounder in medial pivot prostheses than in posterior-stabilized prostheses, but the differences were not significant. The difference in the common logarithm of particle number was significant. The medial pivot prosthesis generated less wear particles than the posteriorstabilized prosthesis, and these findings may have an impact on the incidence of osteolysis and aseptic loosening. PMID- 12771828 TI - An analysis of rotating-platform total knee replacements. AB - Rotating-platform, mobile-bearing total knee replacements have been developed to improve knee kinematics, lower contact stresses on the polyethylene tibial component, minimize constraint, and allow implant self-alignment. The purpose of the current study was to examine some of these parameters. Gait studies during normal gait showed that the stance phase was associated with knee flexion between 8 degrees and 15 degrees. Contact area studies have shown two types of rotating platform total knee replacements, namely gait congruous (congruous only during the stance phase of gait) and totally congruous (congruous up to 90 degrees knee flexion) implants. Knee simulator studies have shown increased gravimetric wear with rotating-platform total knee replacements compared with their fixed-bearing counterparts. Rotate-only implants had less gravimetric wear than rotate and translate rotating-platform total knee replacements. Clinical studies show similar outcomes (knee scores, range of motion, and complications) when rotating platform and fixed-bearing total knee replacements are compared. Although attractive, the benefits of rotating-platform total knee replacements still need to be proven. PMID- 12771829 TI - Gait cycle finite element comparison of rotating-platform total knee designs. AB - Functional load transmission and kinematic performance were compared for standard versus posterior-stabilized versions of a rotating-platform total knee implant, over a standardized loading cycle, using three-dimensional contact finite element analysis. These two design variants differ primarily in terms of the latter's polyethylene insert having a cam that engages with the femoral component during appreciable flexion, thereby inducing femoral component rollback. The finite element model, previously validated experimentally, afforded direct comparisons of anterior lift-off of the insert from the tibial tray, of bearing mobility (insert rotation about the pivot post), of femoral rollback, and of metal-on polyethylene contact stresses at the bearing and backside surfaces of the insert. Both design variants generally performed comparably, exhibiting an internal and external rotation range of approximately 5 degrees, approximately 1.5 mm peak lift-off at the anterior aspect of the insert, and approximately 15 mm of posterior rollback, the respective maxima for both designs occurring at approximately the same instants in the gait cycle. However, the posterior stabilized design had slightly more rollback, and slightly less anterior lift-off and rotation, than did the standard rotating-platform design. Peak polyethylene stresses occurred on the backside of the insert near the posterior edge of the medial compartment, the magnitude being approximately 18% higher for the posterior-stabilized design (21 MPa) than for the standard design. PMID- 12771830 TI - Effect of femoral component design on unresurfaced patellas in knee arthroplasty. AB - Three total knee designs were evaluated to test the hypothesis that femoral component design affects the clinical and mechanical functions of the unresurfaced patella after total knee arthroplasty. Patients with the Ortholoc II, Advantim, and Profix femoral components were followed up for as many as 14 years and revision rate, anterior knee pain, and generalized knee pain were compared. A laboratory protocol was devised to evaluate pressure in the patellofemoral joint of knees from cadavers with a pressure-sensitive transducer using the same three designs at various degrees of knee flexion. Thirty Ortholoc II knee components were followed up for 14 years. Nineteen patients (63%) had severe anterior knee pain and 15 patients (50%) had reoperation to resurface the patella within 2 years. Two hundred one patients (222 knees) with Advantim components were followed up for 10 years and 305 patients (330 knees) with Profix components were followed up for 5 years. No patients with these two knee designs had severe anterior knee pain or reoperation for patellar resurfacing. A significantly higher rate of mild anterior knee pain was seen in the patients with Advantim components than in the patients with Profix components. No apparent relationship was seen between the severity of patellar wear found at the time of surgery and the incidence of anterior knee pain. Patients with rheumatoid arthritis receiving either the Advantim or Profix knee component performed as well as patients with osteoarthritis when the patella was not resurfaced. Pressure was significantly higher in the patellofemoral joints of the laboratory knee specimens with Ortholoc II components than in the specimens with either the Advantim or Profix components. The specimens with Advantim components had significantly higher pressure than did the specimens with normal knees, and the specimens with Profix components differed little from those with normal knees. PMID- 12771831 TI - Femoral component design and patellar clunk syndrome. AB - The effect of patellar design of the femoral component on the prevalence of patellar clunk was examined by comparing 179 consecutive Insall-Burstein II posterior-stabilized total knee replacements with 210 consecutive primary Advanced posterior-stabilized total knee replacements. In the Advanced posterior stabilized knee replacements, the patellofemoral groove had been extended posteriorly 7.5 mm compared with the Insall-Burstein II implant. At a minimum followup of 2 years, the prevalence of patellar clunk syndrome in the patients with a Insall-Burstein II knee replacement was 3.9% (seven of 179) compared with 0% in the patients with an Advanced posterior-stabilized knee replacement. Based on these data, it seems that the design change in patellofemoral groove of the Advanced posterior-stabilized knee replacement has eliminated the problem of patellar clunks. PMID- 12771832 TI - Treatment of the late sequelae of septic arthritis of the hip. AB - Numerous salvage procedures have been proposed for the treatment of late sequelae of septic arthritis of the hip. Despite this, there are no reports in the literature discussing treatment with simultaneous hip reconstruction and femoral lengthening using the hybrid advanced Ilizarov method. The authors reviewed their experience with this technique, and present 15 patients treated between 1982 and 1997. The average age of the patients was 21.1 years and the average limb length discrepancy was 6.5 cm. All the hips were classified according to the classification of Choi et al. The time the external fixator was worn was 225.5 days and the average followup was 108 months. The results were classified based on pain relief, residual deformity, range of motion, Trendelenburg sign, and limb length discrepancy. Ten patients had a good or excellent result, three patients had a fair result, and only two patients had a poor result. Major complications included a common peroneal nerve palsy in one patient, loss of angulation of the proximal femoral osteotomy in two patients, and a mild knee subluxation in three patients all of whom responded to treatment. At the latest followup, 13 patients were satisfied with the treatment, all had returned to their previous occupations, and no patient had a total hip arthroplasty. PMID- 12771833 TI - Alcohol-induced adipogenesis in bone and marrow: a possible mechanism for osteonecrosis. AB - The effect of alcohol on rabbit bone marrow and on the differentiation of mouse bone marrow stromal cells was investigated. Alcohol was administered intragastrically at a dose of 10 mL/kg/day for 1 to 6 months. Alcohol induced a significant increase in serum lipid peroxides, triglyceride, and cholesterol, and a reduction in superoxide dismutase activity. Fatty infiltration in the liver and adipogenesis in bone marrow were found histologically after alcohol administration. Fat cell hypertrophy and proliferation and diminished hematopoiesis in the subchondral area of the femoral head were observed. Triglycerides were deposited in osteocytes, which became pyknotic, and the percentage of empty osteocyte lacunae increased. None of these abnormal changes were detectable in the control group. In the in vitro study, the marrow stromal cells were treated with increasing (0.03, 0.09, and 0.15 mol/L) concentrations of ethanol for 4 to 21 days. Alcohol induced the differentiation of the cells into adipocytes. The number of adipocytes increased with longer durations of exposure to ethanol and with higher concentrations. Cells treated with ethanol also showed diminished alkaline phosphatase activity and expression of osteocalcin. These novel findings indicate that alcohol can directly induce adipogenesis, decrease osteogenesis in bone marrow stroma, and produce intracellular lipid deposits resulting in the death of osteocytes, which may be associated with the development of osteonecrosis, especially in patients with long-term and excessive use of alcohol. PMID- 12771834 TI - Comparison of clinic- and home-based rehabilitation programs after total knee arthroplasty. AB - One hundred sixty patients (mean age, 68 +/- 8 years) having primary total knee arthroplasty were assigned randomly to two rehabilitation programs: (1) clinic based rehabilitation provided by outpatient physical therapists; or (2) home based rehabilitation monitored by periodic telephone calls from a physical therapist. Both rehabilitation programs emphasized a common home exercise program. Before surgery, and at 12 and 52 weeks after surgery, no statistically significant differences were observed between the clinic- and the home-based groups on any of the following measures: (1) total score on the Knee Society clinical rating scale; (2) total score on the Western Ontario and McMaster Universities Osteoarthritis Index; (3) total score on the Medical Outcomes Study Short Form; (4) pain scale of the Knee Society clinical rating scale; (5) pain scale of the Western Ontario and McMaster Universities Osteoarthritis Index; (6) functional scale of the Western Ontario and McMaster Universities Osteoarthritis Index; (7) distance walked in 6 minutes; (8) number of stairs ascended and descended in 30 seconds; and (9) knee flexion range of motion, on either the per protocol or the intent-to-treat or the analyses. After primary total knee arthroplasty, patients who completed a home exercise program (home-based rehabilitation) performed similarly to patients who completed regular outpatient clinic sessions in addition to the home exercises (clinic-based rehabilitation). Additional studies need to determine which patients are likely to benefit most from clinic-based rehabilitation programs. PMID- 12771835 TI - Kinematics of three variations of the Freeman-Samuelson total knee prosthesis. AB - The effect of changes in the articulating surfaces on the kinematics of the Freeman-Samuelson total knee replacement was evaluated. Twenty-two patients (22 knees) (median age, 69 years) were randomized to a standard design with a fixed polyethylene bearing, a design with a mobile bearing, or a new design with a spherical medial femoral condyle with a fixed bearing. The patients were studied with radiostereometry and film-exchangers at 0 degrees nonweightbearing and during active weightbearing extension from 45 degrees to 15 degrees. The center of the tibial tray with a mobile-bearing prosthesis occupied a more anterior position than observed in the designs with a fixed bearing. The medial femoral condyle center had a more posterior position in the standard design than in the design with a spherical medial condyle. From a distally displaced position at 45 degrees, the medial condylar center displaced a mean of 1.8 mm proximally during extension in the standard design and had a constant position in the meniscal design. In the spherical design the medial condyle was displaced proximally at 45 degrees and displaced a mean of 1.6 mm distally during extension. The reduced anteroposterior and proximodistal translations in the meniscal design are compatible with improved congruency between the components. The design with a spherical medial condyle stabilized anteroposterior motions, but showed more pronounced proximodistal displacement medially than did the other two designs. PMID- 12771836 TI - Polyethylene damage on the nonarticular surface of modular total knee prostheses. AB - Modular tibial implants submitted for retrieval analysis were examined for evidence of cold flow, wear, or polyethylene failure. All retrieved components showed areas of cold flow and wear. Significant damage, defined as pitting, gouging, or delamination was observed in 77% of the retrieved implants. Cold flow deformation commonly was observed at the junction of the polyethylene and locking mechanisms or junction of the polyethylene and the edge of the tibial tray. Control implants did not show such areas of damage. Clinical factors such as age, activity status, length of implantation, limb alignment before revision, and thickness of the tibial insert did not correlate with backside wear. No association was found between polyethylene thickness, time of implantation, limb or prosthesis alignment, or activity status or weight of the patient. PMID- 12771837 TI - Blood management after bilateral total knee arthroplasty. AB - Four hundred sixty-one patients who had bilateral one-stage total knee replacements were reviewed to evaluate their blood management. Overall, patients received an average of 2.1 units of autologous blood and 0.9 units of allogenic blood. Seventy-six percent of the patients who preoperatively donated one unit of autologous blood required allogenic blood transfusions compared with 51% of patients who donated two units, 29% of patients who donated three units, and 27% of patients who donated four units of autologous blood. Ninety-eight percent of the patients who did not donate autologous blood required allogenic blood. Donating two units of autologous blood in combination with a perioperative cell saver reduced the incidence of allogenic blood transfusions to 8% but increased the amount of unused autologous blood to 54%. If the indication for wound drainage recovery is guided by the preoperative hematocrit (< or = 40%) or postoperative hemoglobin (> or = 11 mg/dL) the incidence of allogenic blood transfusions decreased to 17% and 13%, respectively and the amount of unused autologous blood decreased to 39% and 30%. There is no statistical difference among the three protocols regarding the need for allogenic blood transfusions and associated costs. Based on this retrospective evaluation the combination of preoperative donation of two units autologous blood and use of a postoperative cell salvage system in all patients is recommended. PMID- 12771838 TI - Failure of operative treatment in a child with osteoporosis-pseudoglioma syndrome. AB - A 6-year-old girl with osteoporosis-pseudoglioma syndrome had operative treatment of a distal femur fracture that failed. Osteoporosis-pseudoglioma syndrome is an autosomal recessive syndrome combining severe premature osteoporosis with a bilateral eye disorder leading to early onset of blindness. Beginning in early childhood, the patient sustained multiple fractures of the left distal femur that were treated nonoperatively. At the age of 5 years the patient had a fracture of the left distal femur with an 80 degrees angulation in the sagittal plane. The patient was treated with internal stabilization because of gross bowing of the femur at the fracture site. Intraoperatively, anatomic reduction was achieved by insertion of a flexible nail but not without some intraoperative problems. Because of the severe osteoporosis, iatrogenic penetration and fracture of the femoral cortex by the nail occurred intraoperatively in the subtrochanteric region. In addition, a hip spica cast was applied. The nail was removed 1 week later. After 6 weeks wearing the hip spica cast, the patient's fracture healed with some shortening but with correction of the angulation in the sagittal plane. Internal stabilization seems to be potentially troublesome in patients with severe forms of this syndrome and severe bony deformities. PMID- 12771839 TI - Multicentric giant cell tumor of bone: a case report and review of the literature. AB - Multicentric giant cell tumor of bone is the rare variant of a lesion that is relatively common in a skeletally mature population. An otherwise healthy 13-year old boy presenting with this entity was followed up for 6 years. During this period, the patient was diagnosed with and treated for six individual lesions. One recurrence required resection, Ilizarov bone lengthening, and subsequent ankle arthrodesis. He remains fully active and free of distant metastasis. PMID- 12771840 TI - Xanthoma of bone: first sign of hyperlipidemia type IIB: a case report. AB - A xanthoma, located in the ulna, not accompanied by the traditional cutaneous and tendinous manifestations (xanthoma and xanthelasma) and with a late onset of alterations in lipid values, was diagnosed in a 56-year-old man. The lesion had a slow but constant growth leading to internal calcifications. Hyperlipidemia Type IIB occurred 15 years after the xanthoma first was detected by radiographs. Therefore, in this patient, xanthoma of bone was the first sign of dyslipidemia. PMID- 12771841 TI - Prospective randomized surgical treatments for calcifying tendinopathy. AB - Thirty-eight patients with chronic calcifying tendinopathy of the shoulder were randomized prospectively into two groups: 19 patients had endoscopic decompression and 19 had open decompression. Thirty-three patients (19 with open and 14 with endoscopic decompression) were available for followup. Clinical and ultrasonographic investigations were done at followup. The purpose of the current study was to evaluate prospectively the clinical and sonographic results after open decompression and after endoscopic decompression. Apportionment of age and gender was similar in both groups. The followup was 15.7 months for patients after open decompression and 17.1 months for patients after endoscopic decompression. The average Patte score was 84.5 points (84.4 for patients who had endoscopic decompression and 84.6 for patients who had open decompression). Corresponding results were found with a Constant and Murley score of 96.6 points (97.6 for patients who had endoscopic decompression and 95.8 for patients who had open decompression). In the endoscopic group physiotherapy was recommended for 5 weeks more than in the open surgery group. Similar results were seen for the incapacity to work in both groups with 4.6 weeks for the patients who had endoscopic surgery and 5 weeks for the patients who had open surgery. No nerve injuries, wound infections, or other postoperative complications were found. Comparison of both surgical methods showed good clinical and score results. Endoscopic and open surgery are equally effective in the treatment of chronic calcifying tendinopathy. PMID- 12771842 TI - Functional free latissimus dorsi muscle flap to the proximal lower extremity. AB - Surgical treatment of lower extremity sarcoma often requires complete resection of muscle compartments resulting in disabling functional loss. Free muscle transfer has been used to restore function of the face and upper extremities, but few reports exist describing functional restoration of a lower extremity. A case report of a 21-year-old man requiring complete resection of the quadriceps musculature with successful functional reconstruction using a free latissimus dorsi muscle flap is described. PMID- 12771843 TI - Treatment of isolated type I open fractures: is emergent operative debridement necessary? AB - A retrospective review was done of treatment of isolated Type I open fractures. Ninety-one patients with isolated Type I open fractures were included in the study. Patients with multiple injuries, gunshot wounds, hand injuries, compartment syndromes, intraarticular fractures, or higher type open fractures were excluded. All patients received antibiotics and were followed up until fracture union. The patients' charts were reviewed for the type of fracture, mechanism of injury, type of treatment, length of hospital stay, and complications encountered, especially infections. There was a 0% incidence of infection in all patients. Only one patient received operative debridement within 12 hours. A prospective randomized study of the treatment of Type I open fractures is needed to determine whether immediate operative debridement is necessary to prevent infection. Immediate operative debridement may not be necessary in the isolated, low-energy Type I open fracture with stable fracture patterns. PMID- 12771844 TI - The use of acrylic bone cement for suture anchoring. AB - Healing of tendon or ligament sutured to bone depends among other parameters on the mechanical stability of the suture fixation in or to the bone. The authors propose a method of anchoring suture material using bone cement as a substitute for conventional suture anchors. Conditions for secure fixation of suture material in bone cement were assessed and the technique of anchoring suture material with acrylic cement in bone was developed. Mechanical testing and microcomputed tomography of the suture-cement-bone compound were done. It was found that the suture always should be knotted before embedding it at least 2 mm deep in the bone cement. The holes drilled into the bone in which the sutures are secured with cement should be at least 3.5 mm in diameter and 10 mm deep; in cortical bone a tapped thread is required. Sutures can be secured safely using cement anchors which provide higher pull-out strength of a factor two to five than conventional metallic suture anchors of comparable size. They also adapt to anatomic situations where conventional anchors cannot be used and are more favorable in osteoporotic bone. Cement anchoring of sutures seems to be a cost effective and valuable alternative when there is poor bone quality or extraordinarily high mechanical load. PMID- 12771845 TI - Calcium sulfate used as bone graft substitute in acetabular fracture fixation. AB - The purpose of this study was to determine the natural history of calcium sulfate pellets implanted during acetabular fracture surgery. The study group consisted of patients sustaining an acetabular fracture with intraarticular comminution or marginal impaction or both in whom calcium sulfate pellets were implanted in lieu of autologous bone graft. Between 1997 and 1999, 32 fractures were treated. Followup adequate to delineate pellet outcome, including radiographs and computed tomography, was obtained in 31 patients. Evaluation of plain radiographs showed that the calcium sulfate pellets became undifferentiated from the surrounding bone at an average of 7 weeks postoperatively. In no case was a residual bony deficit seen. Computed tomography analysis showed that in 22 patients, the pellets essentially had been (> 90%) replaced by bone and in four patients, the majority (> 50%-90%) of the pellets had been replaced by bone. However, in five patients, less than 50% of the pellets had been replaced by bone, including one showing no bony replacement. The common finding in patients with an extensive residual deficit was direct communication of the pellets with the joint space shown on the postoperative computed tomography scan. Patients with the best results had complete containment of the pellets within bone. Therefore, it seems that implanted calcium sulfate pellets in contact with joint synovial fluid are at risk for resorption without significant bony response. If calcium sulfate pellets are to be implanted in a periarticular location, complete bony containment is desirable. Evaluation of the periacetabular bony response requires computed tomography scans, as plain radiographs are inadequate for this purpose. PMID- 12771847 TI - Superior peroneal retinaculoplasty: a surgical technique for peroneal subluxation. AB - A modification of a soft tissue reconstruction procedure for treatment of peroneal tendon subluxation and dislocation is presented. The procedure is based on creating one periosteal flap from the posterior fibula in the region of the peroneal groove. The flap is used to reinforce the superior peroneal retinaculum and prevent anterior displacement of the tendons. This reconstructive procedure has several advantages over existing procedures. The major advantage is that it recreates the normal anatomy of the superior peroneal retinaculum with little disturbance of the surrounding tissues. In addition, the procedure is simple and can be done relatively quickly for patients with acute and chronic subluxation and/or dislocation. Finally, because metallic hardware (such as screws or suture anchors) is not used, artifacts are minimized on magnetic resonance imaging scans. PMID- 12771846 TI - Bioabsorbable miniplating versus metallic fixation for metacarpal fractures. AB - Bioabsorbable implants offer an attractive alternative to metallic implants to stabilize small bone fractures in the hand. Self-reinforced bioabsorbable miniplating for metacarpal fractures was studied in bones from cadavers and compared with standard metallic fixation methods. One hundred twelve fresh-frozen metacarpals from humans had three-point bending and torsional loading after transverse osteotomy followed by fixation using seven methods: (1) dorsal and (2) dorsolateral 2-mm self-reinforced polylactide-polyglycolide 80/20 plating, (3) dorsal and (4) dorsolateral 2-mm self-reinforced poly-L/DL-lactide 70/30 plating, (5) dorsal 1.7-mm titanium plating, (6) dorsal 2.3-mm titanium plating, and (7) crossed 1.25-mm Kirschner wires. In apex dorsal and palmar bending, dorsal self reinforced polylactide-polyglycolide and poly-L/DL-lactide plates provided stability comparable with dorsal titanium 1.7-mm plating. When the bioabsorbable plates were applied dorsolaterally, apex palmar rigidity was increased and apex dorsal rigidity was decreased. Bioabsorbable platings resulted in higher torsional rigidity than 1.7-mm titanium plating and in failure torque comparable with 2.3-mm titanium plating. Low-profile selfreinforced polylactide polyglycolide and poly-L/DL-lactide miniplates provide satisfactory biomechanical stability for metacarpal fixation. These findings suggest that bioabsorbable miniplating can be used safely in the clinical stabilization of metacarpal and phalangeal fractures. PMID- 12771849 TI - Humerus mass in a 25-year-old woman. PMID- 12771848 TI - Painful limp in a 10-year-old boy. PMID- 12771850 TI - Non-invasive exclusion and diagnosis of pulmonary embolism by sequential use of the rapid ELISA D-dimer assay, clinical score and spiral CT. AB - Pulmonary angiography is the gold standard for segmental pulmonary embolism (PE) but no longer for its subsegmental PE, because the inter-observer agreement for angiographically documented subsegmental PE is only 60%. Two non-invasive tools exclude PE with a negative predictive value of > 99%: a normal perfusion lung scan and a normal rapid ELISA VIDAS D-dimer test. The positive predictive value of a high probability ventilation-perfusion lung scan (VP-scan) is only 85% to 87%. The combination of a low clinical score and a non-diagnostic VP-scan safely exclude PE without the need of angiography. The prevalence of PE and that of an alternative diagnosis in symptomatic patients with a non-diagnostic VP-scan are 10% to 20% and 30% to 45%, respectively. Helical spiral computed tomography (CT) detects all clinically relevant PE and a large number of alternative diagnoses in symptomatic patients with a non-diagnostic or high probability VP-scan. The positive predictive value of the spiral CT is > 95%. Single-slice helical CT as the primary diagnostic test in patients with suspected PE in retrospective outcome studies and in prospective multicenter management studies indicate that the negative predictive value of a negative spiral CT preceded or followed by a negative compression ultrasonography (CUS) is > 99%. Therefore, a helical spiral CT can replace both the VP-scan and pulmonary angiography to safely rule in and out PE. A negative rapid ELISA VIDAS D-dimer test result will reduce the need for helical spiral CT by 25% to 35%. PMID- 12771851 TI - High sensitivity C-reactive protein in cardiovascular risk assessment. CRP mania or useful screening? AB - Elevated level of the acute phase reactant C-reactive protein (CRP) is a very sensitive marker of acute inflammatory reactions. Using high sensitivity assays for CRP, recent observations indicate that slightly elevated CRP levels which would be in the normal range of conventional assays are a novel marker for an increased risk for cardiovascular events, especially coronary artery disease and myocardial infarction. Various large scale prospective trials including the Physicians' Health Study and the Women's Health Study revealed that slightly increased hsCRP levels at base line in apparently healthy persons are associated with a 2-fold increase in the risk of a future myocardial infarction. The predictive value of hsCRP was found to be independent from classic risk factors, in particular from elevated serum cholesterol. An increase in hsCRP levels was also associated with a higher risk to develop peripheral artery disease and with a faster progression of carotid artery disease. Until now, treatment with HMG-CoA reductase inhibitors (statins) has been found to be the only medication to consistently decrease hsCRP levels, although by about 15% only. Despite the association of elevated hsCRP levels with future cardiovascular events, routine measurement of hsCRP for cardiovascular risk assessment is currently not recommended because of its low accuracy, the lack of a clear cut-off point for elevated hsCRP levels and the lack of an absolute predictive value. So far, hsCRP remains an interesting potential risk marker for cardiovascular disease whose definite relevance remains to be established. PMID- 12771852 TI - Efficacy of a 6-month treatment with Daflon 500 mg in patients with venous leg ulcers associated with chronic venous insufficiency. AB - AIM: Epidemiological data show that standard compression therapy for leg ulceration in chronic venous insufficiency (CVI) often fails to effectively improve patients' condition. This study assesses the contribution of Daflon 500 mg added to conventional therapy in the healing of hypostatic ulcers of CVI patients. METHODS: Patients of about 65 years were included, with ulcers > or = 2 and > or = 10 cm diameter on 1 or 2 limbs, Doppler ankle/arm pressure index > 0.9, and no recent history of skin graft. Controls (n=68) remained on compression alone while the tested group (n=82) also received Daflon 500 mg 2 tablets/day during 6 months. Treatment could be stopped as soon as the reference ulcer appeared fully healed. Primary endpoints were the rate of healed ulcers and the time to complete healing assessed by planimetry/photography and clinical examination. Variations of the ulcer surface, appearance of the skin, and clinical symptoms of CVI were the secondary criteria. RESULTS: Only 7% of Daflon 500 mg patients necessitated the full 6 month therapy. Whatever the lesion size, from W8 significantly more healed ulcers were observed under Daflon 500 mg (p=0.004), and the ulcer surface was more reduced (p=0.012). For large ulcers, the rate of healing was approximately 2-fold higher with Daflon 500 mg, and the percentage of ulcers healed before W24 was significantly higher (p=0.008). Heavy leg sensation was significantly improved by Daflon 500 mg from W4 (p < 0.05). No treatment-related side effects were reported and the acceptability was considered excellent by 85% of Daflon 500 mg patients. CONCLUSION: Six months of Daflon 500 mg in addition to compression significantly improve some clinical symptoms and accelerate the healing process in patients with ulcerous complications of CVI, with a good acceptability. PMID- 12771853 TI - Midterm experience with the endovascular treatment of isolated iliac aneurysms. AB - AIM: The aim of this retrospective, single institution study was to describe our 4-year experience with the endovascular repair of isolated iliac artery aneurysms. METHODS: Between May 1997 and June 2001, 16 patients (15 males; mean age 64+/-9 years), were treated with covered stent grafts. Twelve of the endovascular procedures were performed under epidural and 4 under local anaesthesia. The percutaneous approach was employed in 13 cases and the femoral artery had to be exposed in 3 cases that demanded simultaneous revascularization of the peripheral circulation (n=2) or required a 16 F sheath to employ a Baxter Lifepath stent graft (n=1). The mean size of the iliac aneurysms was 4.5 cm (range 3.5 to 5.2 cm). Four aneurysms involved the hypogastric ostium in absence of any distal neck. RESULTS: All the patients underwent initially successful endovascular treatment of isolated iliac aneurysms and were followed from 3 to 52 months (mean 18 months). No procedural deaths and no acute or late graft thrombosis occurred. The perioperative complications included 1 dissection of the external iliac artery that required a further endovacular procedure and 1 case of endovascular leak fed to the hypogastric artery. A CT scan 4 months later showed spontaneous thrombosis of aneurysm and no further leakage. Two patients had undergone combinated femoro-popliteal arterial bypass. CONCLUSION: In our early clinical experience the use of self-expandable covered stent graft successful treated isolated iliac artery aneurysms. Endovascular repair is a safe and effective technique with good midterm results in patients at standard and high risk. PMID- 12771855 TI - Increased mast cell infiltration in familial varicose veins: pathogenetic implications? AB - AIM: Increased infiltration of activated mast cells has been recently implicated in the pathophysiology of varicose veins. The aim of the present study was to investigate a possible association between mast cell infiltration of primary varicose veins and clinical features, which could clarify further varicose vein pathophysiology. METHODS: Seventeen patients, operated on for primary varicose veins and greater saphenous vein incompetence, participated in the study. Mast cells, distributed within the adventitia of grossly abnormal segments of the greater saphenous vein and calf varicosities removed during surgery, were identified and measured in stained tissue sections. The mast cell count, expressed as mast cells per 10 high-power fields, was subsequently associated with clinical features, including age, gender, body mass index, familial varicose veins, duration of varicose vein disease and relation to previous pregnancies, leg symptoms and findings on physical examination, clinical class and score of chronic venous insufficiency (CEAP classification). RESULTS: Patients with family history of varicose veins (n=7) had a significantly increased mast cell infiltration (median, interquartile range) of the abnormal venous segments (16, 8.4) in comparison with those (n=10) without such a history (9.2, 7.3), p=0.005. Mast cell infiltration had a significant inverse association with age (r= -0.49, p=0.046), but not with the remaining clinical features. CONCLUSION: Our findings support the hypothesis that the increased mast cell infiltration in varicose veins is not a consequence of venous hypertension. Furthermore, the increased mast cell infiltration in familial varicose veins implies a rather primary role and therefore the presence of a distinct pathophysiology. Further investigation testing the activity of mast cells in cases of family history might reveal another step in the pathogenic mechanism of varicose veins, leading to a more rational treatment. PMID- 12771854 TI - Costs in follow-up of endovascularly repaired abdominal aortic aneurysms. Magnetic resonance imaging with MR angiography versus EUROSTAR protocols. AB - AIM: Endovascular repair of abdominal aortic aneurysms (AAA) necessitates a long term follow-up. These patients are often old and renal insufficiency is not unusual. Cost-effectiveness needs to be addressed in evaluating methods of follow up. The aim of this study was to compare costs of 5 years follow-up with magnetic resonance imaging with contrast enhanced three-dimensional magnetic resonance angiography (MRI/MRA) with follow-up using CT with DSA, or CTA. We also assessed the impact of contrast media induced (CMI) nephropathy on follow-up costs. METHODS: We have implemented Swedish costs of CT with DSA, and CTA on the reported follow-up examinations from the EUROSTAR progress report 2000. The costs of follow-up with CT with DSA, or CTA were compared to a follow-up protocol with MRI/MRA. A cost analysis including a risk analysis of CMI nephropathy was made between MRI/MRA and CT with DSA, or CTA. RESULTS: Excluding the risk of CMI nephropathy, the 5 years follow-up cost in Euro ( ) with MRI/MRA ( 5715) is substantially higher than CT with DSA ( 3 095) or CTA ( 3573). The cost analysis favours MRI/MRA if the risk of CMI nephropathy from CT with DSA, or CTA is more than 5%. CONCLUSION: MRI/MRA can be cost-effective for follow-up of endovascularly repaired AAA depending on the risk of CMI nephropathy for CT with DSA, and CTA. MRI/MRA should be the method of choice for patients with pre existing renal insufficiency. PMID- 12771856 TI - Epidemiological characteristics of acute pulmonary thromboembolism in Japan. AB - AIM: In Japan, acute pulmonary thromboembolism (APTE) is still rare, but the number of patients with APTE has been steadily increasing. It is important for early diagnosis and early management of APTE to recognize epidemiological characteristics of this condition. METHODS: We investigated the epidemiological characteristics of 252 patients with APTE who were admitted to our institutions between 1975 and 2001. APTE was more prevalent in women that in men. It was observed the most in the age group between 50s to 70s, especially in women. Many patients had prolonged immobilization, recent major operation, obesity, or cancer, as risk factors for venous thromboembolism. One hundred and thirty-eight patients developed APTE in hospital; 60 patients were in Department of Internal Medicine, 28 in General Surgery, 15 in Orthopedics, 15 in Obstetrics and Gynecology, and 20 in other services. RESULTS: Among 58 patients with malignancy, 43% had cancers in digestive organs, 21% in gynecological, and 17% in urological. Among 61 patients who were examined for the presence of thrombophilia, 13 patients had inherited thrombophilia (8 protein C deficiency, 4 protein S deficiency, and 1 antithrombin III deficiency) 11 had antiphospholipid antibodies which indicated thrombophilia. Five out of the above 61 patients (8%) had no obvious risk factors including thrombophilia. CONCLUSION: The findings in our patients were almost the same as those in Western patients, except for some points. These results might be useful to establish a preventive approach for APTE in Japan. PMID- 12771857 TI - Prevalence of the G20210A prothrombin gene mutation in Northwestern Greece and association with venous thromboembolism. AB - AIM: The G20210A mutation of the prothrombin gene is a genetic risk factor for venous thromboembolism (VTE). Variability exists in the mutation prevalence in both normal individuals and VTE patients. The aim of this study was to determine the mutation prevalence in Northwestern Greece and evaluate its association with VTE. METHODS: Presence of the G20210A mutation was investigated using DNA analysis in 176 consecutive patients with a history of venous thrombosis or pulmonary embolism and in 300 healthy controls, all Caucasian residents of Northwestern Greece. RESULTS: The mutation was present 12 patients (6.8%) and 8 controls (2.7%). The odds ratio for presence of the mutation versus the normal genotype in VTE was 2.7 (95% CI: 1.1 to 6.7), which was statistically significant. The prevalence of the G20210A prothrombin gene mutation in Northwestern Greece is 2.7% (95% CI: 0.8% to 4.4%) with an allele frequency of 1.3% (95% CI: 0.4% to 2.3%). CONCLUSION: The G20210A mutation of the prothrombin gene is associated with VTE in the Caucasian residents of this geographic region. PMID- 12771858 TI - PGE(1) short term therapy in critical lower limb ischemia. AB - AIM: Our study aims to evaluate the efficiency of short-term therapy with alprostadil (a PGE molecular derivative) on patients affected by critical ischemia of the lower limbs and unsuitable for surgical revascularization. The study was carried out on two groups of patients treated with the traditional long term or a short-term protocol respectively. METHODS: The parameters evaluated and statistically compared to existing studies were, the side effects, subjective pain measured on an analogic scale, objective pain calculated according to analgesic intake, and change in trophic lesions. RESULTS: Our results revealed some differences between the two groups. The manifestation of side effects led to treatment suspension in 8% of long-term therapy cases only. Subjective pain was reduced or disappeared in 83.83% of cases (p<0.001) and there was no statistically significant difference between the two groups. The course of analgesic intake was again similar in both groups. Trophic lesions improved or completely healed in 64.7% (p<0.005) and although a greater tendency towards complete healing was evident in the short-term patients, it was not statistically significant. There was no significant difference between the two groups in the ankle/arm pressure index, but a significant improvement has been observed in 30.88% of cases. The results we obtained from both groups are similar and confirm the valid therapeutic use of alprostadil in patients with peripheral arterial occlusive disease (PAOD). CONCLUSION: Our study reveals the presence of intrinsic advantages to the physician with the short-term schedule, through its higher tolerability, improved and more frequent patient and therapy controls and shorter hospitalization. PMID- 12771860 TI - Elevated plasma fibrinogen levels in patients with essential hypertension are related to vascular complications. AB - AIM: We investigated whether or not fibrinogen is related to the cardiovascular risk profile and complications in hypertensive subjects. METHODS: Plasma fibrinogen and laboratory tests including factor VII, homocysteine and microalbuminuria were evaluated in 127 consecutive hypertensive subjects stratified according to cardiovascular risk. Parameters were age, gender, smoking, cholesterol, diabetes, target organ damage: left ventricular hypertrophy (LVH), carotid atherosclerotic complications and retinical vessels. RESULTS: Fibrinogen levels were significantly different between patients according to risk levels (low 290+/-73, n=20, high 342+/-94 mg/dl, n=39, very high risk 350+/-72, n=29, p=0.01), hypertension grade (II-III) and organ damage. Fibrinogen was significantly higher in patients with more severe carotid atherosclerotic lesions and vascular retinal lesions (grades II-III vs 0 and I). Also in patients, matched for age and sex, without and with carotid atherosclerotic lesions, fibrinogen was significantly higher in the latter group. No significant differences were found on the basis of IVS, creatinine and microalbuminuria. In hypertensive patients, fibrinogen directly correlated with age, by multiple linear regression. In hypertensive patients with diabetes, fibrinogen was significantly higher (466+/-176 mg/dL, n=14) than in those hypertensive without diabetes (333+/-87 mg/dL, n=113, p=0.001) and in all patients there was a a significant correlation (r=0.474, p<0.001) between blood glucose and fibrinogen. CONCLUSION: Hyperfibrinogenemia is a marker of vascular damage and could be an important factor contributing to the evolution of the complications. PMID- 12771859 TI - Spontaneous recanalization of internal carotid artery occlusion evaluated with color flow imaging and contrast arteriography. AB - AIM: In strokes of embolic origin a partial recanalization of the intracranial occluded vessel occurs with a high incidence (as high as 80%). In the literature, we find few cases of revascularization, detected with color flow imaging (CFI) or with arteriography (AGF), at carotid siphon or at the origin of an occluded internal carotid artery (ICA). Up to now there have been no reliable data on the incidence and clinical consequences of SR of an extracranial ICA occlusion. In this case-report we document 8 cases of SR of occluded ICA observed in the last 10 years in our Care Unit. METHODS: We observed 8 complete ICA occlusion at the origin, detected with CFI (8 of 8) and with AGF (7 of 8). All symptomatic patients and 2 of 5 asymptomatic patients underwent CT scan in the acute phase of stroke. All patients underwent CFI follow-up (every 6-12 monhts) to evaluate contralateral CCA and ICA and the presence of new focal neurological symptoms. All patients assumed BMT (antiplatelet or anticoagulant therapy). RESULTS: SR occurred in 6 patients between 24 and 96 months, in 1 patient within 8 months and 1 patient within 6 months from the diagnosis of ICA occlusion. Diagnosis of SR was based in all patients with CFI and in 4 patients confirmed with AGF. Five patients underwent CT scan that excluded haemorrhagic transformation of previous ischemic areas or new ischemic events (2 patients did CT scan only after SR). All patients underwent CFI follow-up in a 3-88 months period. There were no new focal neurological symptoms in 7 of 8 patients, 1 patient presented aspecific neurological symptoms. CONCLUSION: Diagnosing SR of occluded extracranical ICA seems to be more frequent than expected. SR is an event that has to be researched in follow-up of these patients; besides, it seems to have a relatively benign outcome with respect to the onset of new neurological symptoms. PMID- 12771861 TI - The impact of heterozygosity for the factor V Leiden and factor II G20210A mutations on the risk of thrombosis in Greek patients. AB - AIM: There is growing evidence that a number of genetic risk factors predispose independently to venous thrombosis and the coexistence of defective genes is involved in the manifestation and recurrence of thrombotic events. The goal of this study was to examine the efficiency of the selection criteria for performing a genetic test for the factor V G1691A (Leiden) and factor II G20210A mutations. METHODS: Blood samples were drawn from 119 patients referred to us by their physicians. FV and prothrombin (FII) mutations were detected by polymerase chain reaction (PCR) followed by digestion with restriction endonucleases MnlI (FV), HindIII and MspI (FII). RESULTS: Patient carrier frequencies were 16.8% and 10.08% for FV Leiden and FII G20210A, respectively. Heterozygosity for FII G20210A was observed in 10.0% of FV Leiden carriers whereas FV Leiden homozygosity was noted in 1.68% of the patients. Genotype frequencies were in conformity with Hardy-Weinberg equilibrium by the chi square goodness of fit test. CONCLUSION: The obtained data provided a substantial genetic explanation of the thrombotic phenotype in approximately 25% of the patients and thus the physicians selection criteria were sufficient for genetic testing. Furthermore, coinheritance of both genetic defects were significantly associated with increased thrombosis risk and that of recurrent thrombosis. PMID- 12771862 TI - Interrelationship between peripheral arterial occlusive disease, carotid atherosclerosis and flow mediated dilation of the brachial artery. AB - AIM: Atherosclerosis is considered a systemic disease. Therefore, in patients with atherosclerotic disease effects on various sections of the arterial system are expected. The aim of our study was to determine whether patients with evident peripheral arterial occlusive disease (PAOD) of the lower limbs have any subclinical functional or structural arterial wall changes in other sections of the arterial system. METHODS: The study included 54 patients with PAOD, Fontaine stage II and a claudication distance from 50 to 500 m (average 250+/-170 m). Their mean age was 64. None of them had any symptoms or signs of coronary or cerebrovascular atherosclerosis (CVD). The control group consisted of 50 healthy volunteers with a mean age of 64 years without any risk factors of atherosclerosis. In all subjects the carotid intima-media thickness (IMT), was measured, the presence of atherosclerotic plaques in the carotid artery (CA) was registered and the endothelium-dependent dilation capability of the brachial artery (BA) during reactive hyperemia was measured using the B-mode ultrasound technique. RESULTS: The average IMT was significantly greater in PAOD patients than in controls (0.8+/-0.2 mm vs 0.6+/-0.1 mm, p<0.001). In patients atherosclerotic plaques in the CA were also more numerous than in controls (38 vs 4, p<0.001). The IMT of patients was related to body mass index (BMI), ankle brachial pressure index (ABI), LDL cholesterol and to the number of atherosclerotic plaques. In PAOD patients flow-mediated dilation of the BA was significantly lower than in controls (7.2+/-4.9% vs 12.3+/-2.1%, p<0.001). The dilation capability of the BA was linearly related to the BMI, ABI and IMT. CONCLUSION: The results of our study show that PAOD patients without clinical evidence of CVD have morphological changes of the CA, increased IMT and numerous atherosclerotic plaques. Furthermore, in PAOD patients flow-mediated endothelium dependent dilation of the peripheral arteries is decreased. These results support the hypothesis that atherosclerosis is a generalized disease, leading to functional and structural changes in several segments of the arterial system. PMID- 12771863 TI - Off-pump reconstruction of pseudoaneurysm of the descending thoracic aorta. AB - We report 2 cases of postoperative pseudoaneurysm of the descending thoracic aorta, repaired successfully by temporary bypass graft without extracorporeal bypass. One patient presented with a large recurrent pseudoaneurysm that developed 30 years after ligation of a patent ductus arteriosus. The other patient presented with a pseudoaneurysm 18 years after thoracic aortic reconstruction for a traumatic aneurysm. In both cases, a temporary bypass graft was created from a major branch of the aortic arch to the femoral artery prior to definitive reconstruction. The postoperative course was uncomplicated by organ dysfunction or neurologic deficit. Creation of a temporary bypass graft can avoid the need for cardiopulmonary bypass in selected patients. By avoiding heparin use, blood loss is decreased, especially when dense pulmonary adhesions from previous surgery are present. PMID- 12771864 TI - Post-sternotomy internal mammary arteriovenous fistula. AB - Arteriovenous fistula from the mammary artery is a rare complication following cardiac surgery. The fistula usually develops within the first 2 weeks after surgery and is initially asymptomatic. Typically, a continuous machinery murmur is heard along the parasternal border of the chest wall. A patient with an arteriovenous fistula between the right internal mammary artery and mammary vein following a combined aortic valve and coronary bypass operation is described. A transthoracic colour Doppler scan led to the diagnosis of the fistula. Because of potential late complications endovascular embolisation of the fistula was successfully performed. PMID- 12771865 TI - Hypereosinophilia-induced digital necrosis in a smoking patient. AB - Raynaud's phenomenon and digital necrosis of the fingers are rare complications of hypereosinophilia. We report a case of a smoking male who developed Raynaud's phenomenon and digital necrosis of the fingers associated with idiopathic hypereosinophilia with angiographically documented occlusion of small and medium arteries of the extremities. The eosinophils may play a thrombotic role in vascular pathology and eosinophil blood count should be checked when investigating patients with digital gangrene. PMID- 12771867 TI - The operation. PMID- 12771868 TI - Glucose control during cardiac surgery: How sweet it is. PMID- 12771869 TI - Staging of esophageal carcinoma. PMID- 12771870 TI - Should the esophageal cancer staging system be revised? PMID- 12771871 TI - Gene therapy in cardiac surgery: is there a role? PMID- 12771872 TI - Multigene adenoviral therapy for the attenuation of ischemia-reperfusion injury after preservation for cardiac transplantation. AB - OBJECTIVE: The protective effect of adenovirus-mediated ex vivo multigene transfer with superoxide dismutase, a free radical scavenger, and nitric oxide, a vasodilator with anti-inflammatory properties, was examined in the rat heart during experimental ischemia-reperfusion mimicking preservation for cardiac transplantation. METHODS: Donor rat hearts (n = 6 per group) were perfused with solution containing adenoviral vector carrying genes for beta-galactosidase (group A), endothelial nitric oxide synthase (group B), manganese superoxide dismutase (group C), or both endothelial nitric oxide synthase and manganese superoxide dismutase (group D). Hearts were then implanted heterotopically into the abdomens of recipient rats. Four days later, transplanted hearts were collected, connected to a Langendorff perfusion apparatus, and subjected to 6 hours of ischemia followed by 1 hour of reperfusion. Cardiac function was evaluated with an intraventricular balloon at the beginning of Langendorff perfusion and after ischemia-reperfusion. RESULTS: Effective gene transfection was confirmed with X-gal staining in group A hearts. Positive immunoreactivity for endothelial nitric oxide synthase, manganese superoxide dismutase, or both was present predominantly in cardiomyocytes in group B, C, and D hearts. Percentage recovery of preischemic left ventricular developed pressure was 62.1% +/- 7.36% in group A; recoveries were increased to 79.6% +/- 6.4%, 86.8% +/- 9.1%, and 79.4% +/- 6.2% in groups B, C, and D, respectively. CONCLUSION: These results indicate that adenoviral gene transfer of manganese superoxide dismutase and endothelial nitric oxide synthase can attenuate myocardial ischemia reperfusion injury, with the former providing the most significant protection. Combined overexpression of manganese superoxide dismutase and endothelial nitric oxide synthase did not enhance myocardial recovery any further. PMID- 12771873 TI - Continuous insulin infusion reduces mortality in patients with diabetes undergoing coronary artery bypass grafting. AB - OBJECTIVE: Diabetes mellitus is a risk factor for death after coronary artery bypass grafting. Its relative risk may be related to the level of perioperative hyperglycemia. We hypothesized that strict glucose control with a continuous insulin infusion in the perioperative period would reduce hospital mortality. METHODS: All patients with diabetes undergoing coronary artery bypass grafting (n = 3554) were treated aggressively with either subcutaneous insulin (1987-1991) or with continuous insulin infusion (1992-2001) for hyperglycemia. Predicted and observed hospital mortalities were compared with both internal and external (Society of Thoracic Surgeons 1996) multivariable risk models. RESULTS: Observed mortality with continuous insulin infusion (2.5%, n = 65/2612) was significantly lower than with subcutaneous insulin (5.3%, n = 50/942, P <.0001). Likewise, glucose control was significantly better with continuous insulin infusion (177 +/ 30 mg/dL vs 213 +/- 41 mg/dL, P <.0001). For internal comparison, multivariable analysis showed that continuous insulin infusion was independently protective against death (odds ratio 0.43, P =.001). Conversely, cardiogenic shock, renal failure, reoperation, nonelective operative status, older age, concomitant peripheral or cerebral vascular disease, decreasing ejection fraction, unstable angina, and history of atrial fibrillation increased the risk of death. For external comparison, observed mortality with continuous insulin infusion was significantly less than that predicted by the model (observed/expected ratio 0.63, P <.001). Multivariable analysis revealed that continuous insulin infusion added an independently protective effect against death (odds ratio 0.50, P =.005) to the constellation of risk factors in the Society of Thoracic Surgeons risk model. CONCLUSION: Continuous insulin infusion eliminates the incremental increase in in-hospital mortality after coronary artery bypass grafting associated with diabetes. The protective effect of continuous insulin infusion may stem from the effective metabolic use of excess glucose to favorably alter pathways of myocardial adenosine triphosphate production. Continuous insulin infusion should become the standard of care for glycometabolic control in patients with diabetes undergoing coronary artery bypass grafting. PMID- 12771874 TI - Efficacy of esmolol as a myocardial protective agent during continuous retrograde blood cardioplegia. AB - OBJECTIVE: Esmolol, an ultra-short-acting beta-blocker, is known to attenuate myocardial ischemia-reperfusion injury. The aim of this study was to compare the effects of esmolol and potassium on myocardial metabolism during continuous normothermic retrograde blood cardioplegia. METHODS: Forty-one patients operated on for isolated aortic valve stenosis were randomly assigned to continuous coronary infusion with either potassium or esmolol during cardiopulmonary bypass. Myocardial metabolism was assessed by measuring the transmyocardial gradient of oxygen content indexed to left ventricular mass of glucose, lactate, and nitric oxide. To do so, blood samples were simultaneously withdrawn upstream (in the cardioplegia line) and downstream of the myocardium (in the left coronary ostium) 10 and 30 minutes after aortic crossclamping. RESULTS: Although the cardioplegia flow rate and pressure were similar, esmolol markedly reduced the transmyocardial gradient of oxygen content indexed to left ventricular mass compared with potassium: 13 +/- 6 vs 20 +/- 6 mL of oxygen per liter of blood per 100 g of myocardium, respectively, at 10 minutes and 16 +/- 8 vs 24 +/- 8 mL of oxygen per liter of blood per 100 g of myocardium, respectively, at 30 minutes (P =.009). Coronary glucose and lactate transmyocardial gradients were similar in both groups, indicating adequate myocardial perfusion in all patients at all times. In addition, during retrograde cardioplegia, esmolol showed a lower nitric oxide release compared with that caused by potassium (39 +/- 49 micro mol x L(-1) for potassium vs 14 +/- 8 micro mol x L(-1) for esmolol at 10 minutes and 39 +/- 47 micro mol x L(-1) for potassium vs 6 +/- 8 micro mol x L(-1) for esmolol at 30 minutes, P =.05). However, hemodynamic parameters and plasma troponin I levels remained unchanged postoperatively between the 2 types of cardioplegia. CONCLUSION: Esmolol provides potent myocardial protection in hypertrophied hearts, at least in part, by reducing myocardial oxygen metabolism. PMID- 12771875 TI - Early ischemic preconditioning without hypotension prevents spinal cord injury caused by descending thoracic aortic occlusion. AB - OBJECTIVE: Postoperative neurologic deficits after thoracic aortic reconstruction vary widely. Our previous study showed that delayed ischemic preconditioning could prevent spinal cord injury caused by occlusion of the descending thoracic aorta in pigs. We investigated early ischemic preconditioning in the same model. METHODS: Twenty-eight pigs were divided into 4 groups: group 1 (n = 6) underwent a sham operation, group 2 (n = 6) underwent aortic occlusion for 20 minutes, group 3 (n = 8) underwent aortic occlusion for 35 minutes, and group 4 (n = 8) underwent aortic occlusion for 20 minutes and underwent aortic occlusion 80 minutes later without hypotension for 35 minutes. Aortic occlusion was accomplished by using 2 balloon occlusion catheters placed fluoroscopically at T6 to T8 above the diaphragm and at the aortic bifurcation. Neurologic evaluation was performed by an independent observer according to the Tarlov scale (0-4). The lower thoracic and lumbar spinal cords were harvested at 120 hours and examined histologically with hematoxylin-and-eosin stain. Histologic results (number of neurons and grade of inflammation) were scored (0-4) and were similarly analyzed. Statistical analysis was by means of the Kruskal-Wallis test. RESULTS: Group 4 had a better neurologic outcome at 24, 48, and 120 hours in comparison with group 3 (P <.001). The histologic changes were proportional to the neurologic test scores, with the more severe and extensive gray matter damage in animals of group 3 (number of neurons, P <.001; grade of inflammation, P <.001). CONCLUSION: Early ischemic preconditioning without hypotension protects against spinal cord injury after aortic occlusion, as confirmed by using the Tarlov score and histopathology. PMID- 12771877 TI - Efficiency of a gas diffuser and influence of suction in carbon dioxide deairing of a cardiothoracic wound cavity model. AB - OBJECTIVE: In cardiac surgery, insufflation of carbon dioxide is used for deairing of the heart and great vessels. The aim of this study was to assess a new insufflation device for efficient deairing and to study the influence of suction. METHODS: We measured the content of remaining air at two positions in the cardiothoracic wound model. A new insufflation device, a gas diffuser, was compared with a conventional 0.25-inch tube. Carbon dioxide flow (5 and 10 L/min) and suction (0, 1.5, 10, and 25 L/min) were varied. Suction was studied in combination with the gas diffuser. RESULTS: With the tube the median air content in the wound model was 19.5% to 51.7% at the studied carbon dioxide flows, whereas with the gas diffuser the median air content was no greater than 1.2% at 5 L/min and no greater than 0.31% at 10 L/min (P <.001). When suction of 1.5 L/min was applied, the median air content in the model remained low (/=40 neonates), and procedural variables (shunt originating from aorta, longer circulatory arrest time, and management of the ascending aorta). Of neonates undergoing cavopulmonary shunt, 91% had reached a subsequent transition state by 6 years after cavopulmonary shunt, consisting of Fontan operation (79%), death (9%), or cardiac transplantation (3%). Risk factors for death occurring before subsequent transition included younger age at cavopulmonary shunt and need for right atrioventricular valve repair. CONCLUSIONS: Competing risks analysis defines the prevalence of the various outcomes after Norwood operation and predicts improved outcomes with successful modification of controllable risk factors. PMID- 12771882 TI - Nitric oxide synthase expression by pulmonary arteries: a predictive marker of Fontan procedure outcome? AB - OBJECTIVES: We retrospectively analyzed lung biopsy specimens from patients who underwent the Fontan procedure to identify predictive markers of outcome. METHODS: We studied the intra-acinar pulmonary arteries present in lung biopsy specimens from 17 patients undergoing the Fontan procedure. We evaluated both their morphology and their expression of endothelial nitric oxide synthase and endothelin 1. We compared these data with those of 6 patients who died of no pulmonary cause (control group). RESULTS: Eight patients had a good surgical outcome (group 1). Their distal arteries were thin and weakly expressed endothelin 1 and endothelial nitric oxide synthase. The procedure failed in 9 patients (group 2). Their distal arteries displayed muscle extension with an increased wall thickness (P <.01 vs group 1). Their endothelin 1 expression remained low (not significant vs group 1). By contrast, endothelial nitric oxide synthase was markedly overexpressed (P <.001 vs group 1). CONCLUSION: Distal pulmonary arteries of patients in whom the Fontan procedure failed exhibited a markedly increased wall thickness and a clear endothelial nitric oxide synthase overexpression. In addition to giving clues to the pathogenesis of the procedure's failure, our study might help to define reliable predictive markers of its outcome. PMID- 12771883 TI - Role of clinically determined depth of tumor invasion in the treatment of esophageal carcinoma. AB - OBJECTIVE: We sought to evaluate the effectiveness of clinical staging of depth of tumor invasion (cT), the relationship of cT to survival, the benefits of downstaging cT, and the role of cT in treatment decisions. METHODS: The accuracy of determining T by means of endoscopic ultrasonography and the relationship of cT to survival were assessed in 209 patients undergoing esophagectomy alone for esophageal carcinoma. The benefit of downstaging cT was assessed in 128 patients undergoing induction therapy and esophagectomy. The role of cT in treatment decisions was determined by integrating these results with the results of previous work. RESULTS: Compared with pathologic T (pT), cT was 87% accurate, 82% sensitive, 91% specific, 89% positively predictive, and 86% negatively predictive of tumors confined to (< or =T2) or invading beyond (>T2) the esophageal wall. In cN0, increasing cT was predictive of progressively poorer survival. For each category of pT N0, cT accurately predicted survival, except for pT3, which was underestimated (P <.0001). In cN0, downstaging by induction therapy was beneficial only if tumors invaded beyond the wall (> or =cT3, P =.0003). In cN1, it was beneficial only when downstaging was synchronous in cT3/T4 (P <.001). CONCLUSIONS: cT should be the principal determinant of treatment in cN0. In cN0, if endoscopic ultrasonography identifies tumors of greater than cT2, multimodality therapy should be considered. However, only when cT3/T4 tumors are downstaged to pT2 or less will patients benefit, but their survival will not equal that of patients with tumors of cT2 or less having esophagectomy alone. If endoscopic ultrasonography identifies tumors of cT2 or less, esophagectomy alone should be used because induction therapy might adversely affect survival. PMID- 12771884 TI - Refining esophageal cancer staging. AB - OBJECTIVE: Cancer staging is dynamic, reflecting accrual of knowledge and experience in treatment. The objectives of this study were to assess current esophageal cancer staging and to determine whether refinements of classification and stage grouping are necessary. METHODS: From 1983 through November 2000, 480 patients underwent esophagectomy without induction therapy. Depth of tumor invasion (T), regional lymph node status (N), distant status (M), number of metastatic regional lymph nodes, and histopathologic type and grade were subjected to survival-tree analysis, multivariable Cox and hazard function analysis, and residual misclassification risk analysis. RESULTS: Inhomogenity of survival was found within and lack of distinction was found between current American Joint Committee on Cancer staging groups, supporting the need for refinement. T1 and N1 were redefined on the basis of survival differences. T1a is intramucosal cancer, T1b is submucosal cancer (P =.008), N1 is 1 or 2 metastatic regional lymph nodes, and N2 is 3 or more metastatic regional lymph nodes (P =.01). Current subclassification of M1 is not warranted (P =.9). Histopathologic type (P =.17) and grade (P =.3) minimally refined staging. Reassignment of staging groups constrained by American Joint Committee on Cancer definitions of stages 0 and IV produced less monotonic, distinctive, and homogeneous survival than free assignment of staging groups. CONCLUSIONS: Current American Joint Committee on Cancer staging of esophageal cancer is inadequate. Refinement requires redefinition of T1, N1, and M1 classifications. Stage grouping within the constraints of American Joint Committee on Cancer definitions produces less accurate prognosis than free assignment based on survival data. PMID- 12771885 TI - Transthoracic versus transhiatal esophagectomy: a prospective study of 945 patients. AB - OBJECTIVE: Debate continues as to whether transhiatal esophagectomy results in lower morbidity and mortality than transthoracic esophagectomy. Most data addressing this issue are derived from single-institution studies. To investigate this question from a nationwide multicenter perspective, we used the Veterans Administration National Surgical Quality Improvement Program to prospectively analyze risk factors for morbidity and mortality in patients undergoing transthoracic esophagectomy or transhiatal esophagectomy from 1991 to 2000. METHODS: Univariate and multivariate analyses were performed on 945 patients (mean age, 63 +/- 10 years). There were 562 transthoracic esophagectomies and 383 transhiatal esophagectomies in 105 hospitals, with complete 30-day outcomes recorded. RESULTS: There were no differences in recorded preoperative variables between the groups that might bias any comparisons. Overall mortality was 10.0% (56/562) for transthoracic esophagectomy and 9.9% (38/383) for transhiatal esophagectomy (P =.983). Morbidity occurred in 47% (266/562) of patients after transthoracic esophagectomy and in 49% (188/383) of patients after transhiatal esophagectomy (P =.596). Risk factors for mortality common to both groups included a serum albumin value of less than 3.5 g/dL, age greater than 65 years, and blood transfusion of greater than 4 units (P <.05). When comparing transthoracic esophagectomy with transhiatal esophagectomy, there was no difference in the incidence of respiratory failure, renal failure, bleeding, infection, sepsis, anastomotic complications, or mediastinitis. Wound dehiscence occurred in 5% (18/383) of patients undergoing transhiatal esophagectomy and only 2% (12/562) of patients undergoing transthoracic esophagectomy (P =.036). CONCLUSIONS: These data demonstrate no significant differences in preoperative variables and postoperative mortality or morbidity between transthoracic esophagectomy and transhiatal esophagectomy on the basis of a 10-year, prospective, multi-institutional, nationwide study. PMID- 12771886 TI - Clinical implications of p53 tumor suppressor gene mutation and protein expression in esophageal adenocarcinomas: results of a ten-year prospective study. AB - OBJECTIVE: This study was undertaken to characterize the spectrum of p53 alterations (mutations and protein expression) in surgically resected esophageal adenocarcinomas, and to correlate molecular alterations with clinicopathologic findings and outcome. METHODS: Between 1991 and 2001, 91 consecutive patients with esophageal adenocarcinomas underwent subtotal esophagectomy. No patient received induction therapy. Strict clinicopathologic criteria were used to define primary esophageal adenocarcinomas. Genomic DNA was extracted from esophageal tumors, each matched with histologically normal esophageal epithelium (internal control) from the resection margin. Polymerase chain reaction was used to amplify p53 exons 4 through 10. Mutations were studied by single-strand conformation polymorphism analysis and direct DNA sequencing. Immunohistochemical testing (monoclonal antibody DO7) was used to evaluate p53 protein distribution. RESULTS: Five-year overall survival was 27.3%. No p53 alterations (mutations and/or protein overexpression) were found in normal esophageal epithelium. A total of 57.1% (n = 52) of tumors had p53 alterations (mutations and/or protein overexpression), which on univariate analysis were associated with poor tumor differentiation (P =.001), advanced pTNM stage (P =.009), and number of involved lymph nodes (0, 1-3, >3; P =.04). Patients with p53 alterations had significantly reduced 5-year overall survival relative to patients with wild-type p53 (15% vs 46%; P =.004). The p53 mutations were predominantly G:C to A:T transitions at CpG dinucleotides (52.2%, 24/46) CONCLUSIONS: We conclude that p53 alterations (mutations and/or protein overexpression) are a predictor of reduced postoperative survival after surgical resection of esophageal adenocarcinomas and that p53 may be a clinically useful molecular marker for stratifying patients in future clinical trials. Patterns of p53 mutations suggest endogenous mutational mechanisms. PMID- 12771887 TI - Enhancement of depsipeptide-mediated apoptosis of lung or esophageal cancer cells by flavopiridol: activation of the mitochondria-dependent death-signaling pathway. AB - OBJECTIVE: Treating cancer cells with depsipeptide, a novel antitumor agent currently in a phase II clinical trial, causes potent upregulation of p21/WAF1 expression and cell arrest at G1 and G2 checkpoints. p21/WAF1 upregulation, however, impedes the ability of depsipeptide to induce significant apoptosis. This study was designed to determine whether flavopiridol, a synthetic cyclin dependent kinase inhibitor known to inhibit p21 expression in tumor cells, could enhance depsipeptide-mediated apoptosis in cultured lung and esophageal cancer cells. METHODS: Lung or esophageal cancer cells were exposed to depsipeptide, flavopiridol, or a combination of depsipeptide and flavopiridol. Cytotoxicity and apoptosis were quantitated by means of (4,5-dimethylthiazo-2-yl)-2,5-diphenyl tetrazolium bromide and terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling-based assays, respectively. Cytosolic cytochrome c levels, caspase 9 activity, mitochondrial membrane depolarization, and dependence of apoptosis on caspase 9 in treated cells were studied to determine the role of the mitochondria in mediating apoptosis induced by this drug combination. RESULTS: Flavopiridol completely abolished depsipeptide-mediated dose-dependent upregulation of p21/WAF1 expression. Combining flavopiridol with depsipeptide resulted in a 3- to 8-fold reduction of depsipeptide inhibitory concentration of 50% values that was closely paralleled by synergistic enhancement of apoptosis (4- to 10-fold higher than levels of cell death induced by either drug alone) in all cancer cell lines. The essential role of mitochondria in mediating cell death was indicated by robust translocation of cytochrome c from the mitochondria into the cytosol, 2.5- to 5-fold activation of caspase 9, severe disruption of mitochondrial inner membrane potential, and complete inhibition of apoptosis by the selective caspase 9 inhibitor. More important, this drug combination was not toxic to primary normal epithelial cells derived from the airway or skin. CONCLUSION: The depsipeptide plus flavopiridol combination exhibits powerful and selective cytocidal activity against cancer but not normal cells. Apoptosis induced by this combination is mediated by the mitochondria-dependent death pathway. PMID- 12771888 TI - Late outcomes of mitral valve repair for floppy valves: Implications for asymptomatic patients. AB - OBJECTIVES: We sought to evaluate the long-term results of mitral valve repair in patients with mitral regurgitation caused by floppy mitral valves and compare the outcomes of asymptomatic patients with those of symptomatic patients. METHODS: A retrospective review of 488 consecutive patients who had mitral valve repair for floppy mitral valve disclosed 199 patients who were asymptomatic or had minimal symptoms and 289 patients who were symptomatic at the time of the operation. Asymptomatic patients were younger, had better ventricular function, had a lower incidence of coronary artery disease, and had higher rates of atrial fibrillation than symptomatic patients. RESULTS: Survival at 15 years was 61% for all patients. Survival was 76% for asymptomatic patients, which was identical to that for the general population matched for age and sex, whereas the survival of symptomatic patients was 53% and significantly lower than that of the general population. Cox regression analyses validated by means of bootstrap methodology identified the following predictors of late death: age by increments of 5 years (risk ratio = 1.2), New York Heart Association functional classes 3 and 4 (risk ratio = 3.0), left ventricular ejection fraction of less than 40% (risk ratio = 2.7), preoperative stroke or transient ischemic attack (risk ratio = 3.1), previous cardiac operation (risk ratio = 4.6), and severe chronic obstructive pulmonary disease (risk ratio = 3.1). Freedom from reoperation at 15 years was 91%, and it was similar for asymptomatic and symptomatic patients. Freedom from mitral regurgitation of greater than 2+ at 15 years was 85% for all patients, 96% for asymptomatic patients, and 76% for symptomatic patients. CONCLUSIONS: This study supports the recommendation of surgical intervention in asymptomatic patients with mitral regurgitation caused by a floppy mitral valve if mitral valve repair is feasible and associated with low operative mortality and morbidity. PMID- 12771889 TI - Ruptured bronchial artery aneurysm associated with sarcoidosis. PMID- 12771890 TI - Thoracic splenosis: mimicry of a neurogenic tumor. PMID- 12771891 TI - Thoracosternotomy for off-pump coronary artery bypass. PMID- 12771892 TI - Bilateral diaphragmatic paralysis after aortic surgery with topical hypothermia: Ventilatory assistance by means of nasal mask bilevel positive pressure. PMID- 12771893 TI - Intracoronary artery shunt: an assessment of possible coronary artery wall damage. PMID- 12771895 TI - Combined heart and liver transplantation for familial amyloidotic polyneuropathy. PMID- 12771894 TI - Robotically assisted division of a vascular ring in children. PMID- 12771896 TI - Simultaneous biventricular pacemaker implantation for a surgical case of aortic regurgitation with severe left ventricular dysfunction and left bundle branch block. PMID- 12771897 TI - Indications for gadolinium for coronary angiography. PMID- 12771898 TI - Risk adjustment in analysis of surgery for congenital heart disease. PMID- 12771899 TI - Ranking institutions. PMID- 12771900 TI - Symmetry aortic connector system. PMID- 12771901 TI - Symmetry aortic connector system. PMID- 12771902 TI - Does normothermic cardiopulmonary bypass influence clinical outcomes, cytokine production, and in vitro platelet function? PMID- 12771903 TI - An economic evaluation of lung transplantation. PMID- 12771904 TI - To close or not to close? PMID- 12771905 TI - Which cell dose supports motor neurons' survival? PMID- 12771906 TI - A gauze sponge cannot act as a gas diffuser in cardiac surgery when it gets wet. PMID- 12771907 TI - Thromboembolic complications after beating heart coronary artery bypass operations. PMID- 12771908 TI - Single fiber skeletal muscle transplantation or purified myoblast engraftment? PMID- 12771909 TI - Large cell neuroendocrine carcinoma of the lung: what we have to do. PMID- 12771910 TI - Tuberculous aneurysms of the aorta. PMID- 12771911 TI - To the editor. PMID- 12771913 TI - Prophylactic options in patients with 5-fluorouracil-associated cardiotoxicity. AB - At present, the various mechanisms involved in 5-fluorouracil (5-FU)-correlated cardiotoxicity remain to be elucidated and a universally accepted prophylaxis or treatment for this specific toxicity is not available. Although it may improve time to progression, survival and clinical benefit, a 5-FU-based regimen usually has to be discontinued if a patient experiences cardiotoxicity. Here, we describe our experience with three cases of 5-FU-associated cardiotoxicity. The angina like pain that appeared approximately 95 h after beginning 5-FU therapy was apparently independent of the drug's administration modality. In the two patients receiving 5-FU 12-h flat continuous infusion from 22.00 to 10.00 h (5-FU 12-h c.i.) in combination with other drugs, the dose of 5-FU was reduced by 10-20% and patients received prophylactic transepidermal nitroglycerin. In the third patient, 5-FU administration modality was changed and prophylactic therapy was not given. By taking these precautions, the patients no longer complained of anginal pain and none of them discontinued chemotherapy. PMID- 12771912 TI - Cancer and the FRA3B/FHIT fragile locus: it's a HIT. AB - The FHIT gene encompassing the most active common human chromosomal fragile region, FRA3B, was discovered in 1996 and proposed as a tumour suppressor gene for important human cancers. Seven years and more than 350 reports later, early questions concerning its tumour suppressor role have been answered. Recent studies on the role of Fhit loss in major types of human cancers report association with high proliferative and low apoptotic indices, node positivity, loss of mismatch repair protein, likelihood of progression and reduced survival. PMID- 12771914 TI - Analysis of the time course and prognostic factors determining toxicity due to infused fluorouracil. AB - This study used a prospectively managed clinical database in order to identify 1470 patients with gastrointestinal cancers receiving protracted venous infusion (PVI) fluorouracil (5FU). It aimed to determine the time course of toxicity due to PVI 5FU and to analyse factors predicting toxicity. The initial development of stomatitis occurred more rapidly than diarrhoea or palmar plantar erythema (PPE). The percentage of patients with National Cancer Institute Common Toxicity Criteria (CTC) grade 2 or worse PPE peaked at 9% between weeks 8 and 17, whereas this peak occurred earlier for stomatitis and diarrhoea. The development of CTC grade 1 toxicity in the first 28 days after commencement of chemotherapy was classified as early grade 1 toxicity. Multivariate Cox regression analysis showed that female sex, better performance status, elevated bilirubin, early grade 1 PPE and early grade 1 diarrhoea were independent prognostic factors for the development of CTC grade 2 or worse PPE (P<0.01). Female sex, increased age, elevated alanine transaminase and urea and early grade 1 PPE were significant independent prognostic factors for the development of CTC grade 2 or worse stomatitis (P<0.01). Early CTC grade 1 diarrhoea predicted CTC grade 2 or worse diarrhoea (P<0.01). Older, female patients with good performance status and impaired liver and renal function who develop early grade 1 PPE alone or in combination with diarrhoea are at highest risk of subsequently developing grade 2 or worse PPE or stomatitis during treatment with PVI 5FU. Reduction of infused 5FU dose should be considered for these patients. Such an approach could both reduce severe toxicity owing to chemotherapy and minimise treatment delays, and should be evaluated prospectively. PMID- 12771915 TI - IPM chemotherapy in cytokine refractory renal cell cancer. AB - Renal cell carcinoma (RCC) is notoriously chemoresistant. Current management of metastatic disease usually includes immunological agents of which the most clearly evaluated is alpha interferon. Following the failure of such agents no clear second-line therapy exists. The use of a novel combination of cisplatin, irinotecan and mitomycin may offer some palliative benefit in this situation. Thirty-three patients with cytokine refractory RCC and documented progression and documented active progressive disease with performance status 0-3 were enrolled. Therapy consisted of cisplatin 40 mg m(-2) on day 1 and day 15, irinotecan 100 mg m(-2) on day 1 and day 15, and mitomycin 6 mg m(-2) on day 1 of a 28-day cycle. The results showed that one patient (3%) had a partial response, eight (24%) had minor responses and nine (27%) had stable disease, overall 61% had symptomatic responses. Quality-of-life (QOL) assessment did not change significantly during therapy. Seventy-one percent of those who had primary refractory disease to cytokine therapy subsequently responded to IPM. The median progression-free interval was 4.8 months in this cohort on chemotherapy, compared to 3.9 months with their previous cytokine treatment. In conclusion, IPM produced symptomatic relief for a majority of patients with cytokine refractory RCC without any deterioration in QOL. Disease stabilisation on radiological assessment and symptomatic improvement were associated with prolonged survival. A degree of non crossresistance to cytokine therapy was seen. IPM may be considered in patients with renal cancer following failure of cytokines. PMID- 12771916 TI - Correlation of elevated level of blood midkine with poor prognostic factors of human neuroblastomas. AB - The heparin-binding growth factor midkine (MK) is the product of a retinoic acid responsive gene, and is implicated in neuronal survival and differentiation, and carcinogenesis. We previously reported that MK mRNA expression is elevated in neuroblastoma specimens at all stages, whereas pleiotrophin, the other member of the MK family, is expressed at high levels in favourable neuroblastomas. As MK is a secretory protein, it can be detected in the blood. Here, we show a significant correlation of the plasma MK level with prognostic factors of neuroblastomas. The plasma MK level was determined in 220 patients with neuroblastomas, and compared with that in children without malignant tumors (n=17, <500 pg ml(-1)). The plasma MK level became significantly elevated with advancing stages (stage 1: 445 pg ml( 1) (median), n=73; stage 2: 589, n=39; stage 3: 864, n=40; stage 4: 1445, n=56; and stage 4S: 2439, n=12). More importantly, a higher MK level was strongly correlated with poor prognostic factors: over 1 year of age (P=0.0299), MYCN amplification (P<0.0001), low TrkA expression (P=0.0005), nonmass screening, sporadic neuroblastomas (P<0.0001), and diploidy/tetraploidy (P=0.0007). Thus, these results demonstrate that the plasma MK level is a good marker for evaluating the progression of neuroblastomas. Moreover, considering the ability of antisense MK oligodeoxyribonucleotide to suppress tumour growth of colorectal carcinoma cells in nude mice, as recently reported, the present study suggests that MK is a possible candidate molecular target for therapy for neuroblastomas. PMID- 12771917 TI - Expression of costimulatory molecules in human neuroblastoma. Evidence that CD40+ neuroblastoma cells undergo apoptosis following interaction with CD40L. AB - Tumour cells display low to absent expression of costimulatory molecules. Here, we have investigated the expression of costimulatory molecules (CD40, CD80, CD86, PD-1L, B7H2, OX40L and 4-1BBL) in human neuroblastoma (NB) cells, since virtually no information is available on this issue. Both established NB cell lines and primary tumours were tested by RT-PCR and flow cytometry. Neuroblastoma cell lines expressed the transcripts of all costimulatory molecule genes, but not the corresponding proteins. Culture of NB cell lines with human recombinant (r)IFN gamma induced surface expression of CD40 in half of them. Primary NB cells showed CD40, CD80, CD86, OX40L, 4-1BBL, but not PD-1L and B7H2, mRNA expression. Surface CD40 was consistently detected on primary NB cells by flow cytometry. Interferon gamma gene-transfected NB cells expressed constitutively surface CD40 and were induced into apoptosis by incubation with rCD40L through a caspase-8-dependent mechanism. CD40 may represent a novel therapeutic target in NB. PMID- 12771918 TI - Long-term outcome of medullary thyroid carcinoma in patients with normal postoperative medical imaging. AB - Imaging-detected relapses are observed in a significant proportion of patients with medullary thyroid carcinoma (MTC) with normal postoperative imaging studies. The aim of this study was to search for prognostic factors of imaging-detected relapse. This retrospective study was performed in 63 consecutive MTC patients with normal postoperative medical imaging. After surgery, the basal calcitonin (CT) level was undetectable in 35 patients and elevated in 28. During follow-up, 18 patients developed a clinical or imaging-detected relapse (29%) in the neck and/or at distant sites: 15 had an elevated postoperative basal CT level and three had an undetectable postoperative basal CT level. At multivariate analysis, the significant parameters predictive of imaging-detected relapse were the postoperative plasma CT level and the tumour extension (pT). The 3- and 5-year relapse-free survival rates were 94 and 90% in patients with an undetectable postoperative basal CT level, and 78 and 61% in patients with a detectable basal CT level (P<0.05). The 3- and 5-year relapse-free survival rates were 92 and 85% in the pT1-3 patients, and 57 and 46% in the pT4 patients (P<0.01). These results show that postoperative CT level and tumour extension are critical prognostic factors for the identification of patients at a high risk of relapse. PMID- 12771919 TI - The LIM-only protein, LMO4, and the LIM domain-binding protein, LDB1, expression in squamous cell carcinomas of the oral cavity. AB - Carcinoma cells can lose their epithelial cell characteristics and dedifferentiate into a fibroblast-like cell during progression of a neoplasm. Aberrant expression of oligomeric transcriptional complexes contributes to progression of carcinomas. Although individual transcription factors initiating progression remain unknown, LIM-only protein (LMO) and LIM-domain binding protein (LDB) negatively regulate breast carcinoma cell differentiation. In this study, we investigated the expression of LMO4 and LDB in squamous cell carcinomas of the oral cavity. LMO4 mRNA was amplified in four of six carcinoma tissues and eight of 12 carcinoma cell lines, and LDB1 in three carcinoma tissues and 11 cell lines examined. Immunoprecipitation studies revealed that LMO4 and LDB1 interact with each other in the nuclear milieu of the carcinoma cells indicating the presence of an LMO4-LDB1-mediated transcription complex. Both LMO4 and LDB1 proteins were preferentially localised in the nuclei of carcinoma cells at the invasive front and the immunoreactivity was increased in less-differentiated carcinoma tissues (P<0.01). Carcinoma cells metastasised to the cervical lymph nodes with increased immunoreactivity compared to the primary site of neoplasm (P<0.05). These data suggest that the LMO4-LDB1 complexes may be involved in carcinoma progression possibly through dedifferentiation of squamous carcinoma cells of the oral cavity. PMID- 12771920 TI - Prognostic significance of circumferential resection margin involvement following oesophagectomy for cancer. AB - The factors affecting long-term survival following oesophagectomy for oesophageal cancer are poorly understood. We examined the significance of microscopic tumour involvement at the circumferential resection margin (CRM) on postoperative survival following oesophagectomy. The case notes of 329 patients who underwent a potentially curative oesophagectomy for squamous or adenocarcinoma were reviewed retrospectively. As part of the procedure, all patients underwent an en-bloc resection of their periesophageal tissue. The presence of tumour either at, or within, 1 mm of the CRM was recorded and correlated with their TNM and survival data. A total of 67 patients (20%) were noted to have a positive CRM, of which 40 cases (12%) had tumour at the resection margin and the remainder had tumour within 1 mm of the margin. Univariate analysis showed no statistically significant association between survival and either category of CRM involvement. Multivariate analysis showed that only T-stage, nodal status and tumour grade were prognostic markers. In conclusion, the presence of microscopic tumour at the CRM following an en-bloc oesophagectomy is not a significant prognostic marker. PMID- 12771922 TI - Absence of cyclin D2 expression is associated with promoter hypermethylation in gastric cancer. AB - Expression of cyclin D2 is absent in 30-70% of gastric cancers. We investigated the role of promoter hypermethylation in the transcriptional silencing of cyclin D2 in five gastric cell lines and 47 primary gastric carcinomas. CpG island methylation status of the cyclin D2 gene was studied by methylation-specific polymerase chain reaction and bisulphite sequencing. RNA and protein expression was analysed by reverse transcription-PCR and Western blot, respectively. Dense methylation of cyclin D2 was detected in three cell lines (KATOIII, AGS and NCI N87), which also lacked cyclin D2 mRNA and protein expression. Bisulphite DNA sequencing revealed that loss of cyclin D2 expression was closely associated with the density of methylation in the promoter region. Treatment with DNA methyltransferase inhibitor, 5-aza-2'-deoxycytidine, restored the cyclin D2 expression level in methylated gastric cells. Among the 47 primary gastric cancers, cyclin D2 hypermethylation was detected in 23 (48.9%) cases. None of the 23 normal gastric biopsies from noncancer patients showed hypermethylation. Hypermethylation was associated with loss of mRNA (P&<0.001) and protein (P=0.006) expressions. Our study showed that cyclin D2 hypermethylation is associated with loss of cyclin D2 expression in a subset of gastric cancers, which may suggest an alternative gastric carcinogenesis pathway in the absence of cyclin D2 expression. PMID- 12771921 TI - Matrix metalloproteinases 2 and 9 (gelatinases A and B) expression in malignant mesothelioma and benign pleura. AB - Matrix metalloproteinases (MMPs), in particular the gelatinases (MMP-2 and -9), play a significant role in tumour invasion and angiogenesis. The expression and activities of MMPs have not been characterised in malignant mesothelioma (MM) tumour samples. In a prospective study, gelatinase activity was evaluated in homogenised supernatants of snap frozen MM (n=35), inflamed pleura (IP, n=12) and uninflammed pleura (UP, n=14) tissue specimens by semiquantitative gelatin zymography. Matrix metalloproteinases were correlated with clinicopathological factors and with survival using Kaplan-Meier and Cox proportional hazard models. In MM, pro- and active MMP-2 levels were significantly greater than for MMP-9 (P=0.006, P<0.001). Active MMP-2 was significantly greater in MM than in UP (P=0.04). MMP-2 activity was equivalent between IP and MM, but both pro- and active MMP-9 activities were greater in IP (P=0.02, P=0.009). While there were trends towards poor survival with increasing total and pro-MMP-2 activity (P=0.08) in univariate analysis, they were both independent poor prognostic factors in multivariate analysis in conjunction with weight loss (pro-MMP-2 P=0.03, total MMP-2 P=0.04). Total and pro-MMP-2 also contributed to the Cancer and Leukemia Group B prognostic groups. MMP-9 activities were not prognostic. Matrix metalloproteinases, and in particular MMP-2, the most abundant gelatinase, may play an important role in MM tumour growth and metastasis. Agents that reduce MMP synthesis and/or activity may have a role to play in the management of MM. PMID- 12771923 TI - Activation of the Epstein-Barr virus lytic cycle by the latex of the plant Euphorbia tirucalli. AB - Exposure to the plant Euphorbia tirucalli has been proposed to be a cofactor in the genesis of endemic Burkitt's lymphoma (eBL). The purpose of this study was to examine the effects of unpurified E. tirucalli latex on Epstein-Barr virus (EBV) gene expression. A Burkitt lymphoma cell line was treated with varying dilutions of the latex and the effects on EBV gene expression were measured. We observed that the latex was capable of reactivating the EBV lytic cycle in a dose dependent manner and at dilutions as low as 10(-6). Simultaneous treatment of cells with E. tirucalli latex and the protein kinase C inhibitor 1-(5 isoquinolinesulphonyl)-2-methylpiperazine dihydrochloride blocked lytic cycle activation. These data suggest that environmental exposure to the latex of E. tirucalli could directly activate the EBV lytic cycle and provide further evidence of a role for E. tirucalli in the aetiology of eBL. PMID- 12771924 TI - Inclusion of HPV testing in routine cervical cancer screening for women above 29 years in Germany: results for 8466 patients. AB - In a prospective cohort study 8466 women attending routine cervical cancer screening were recruited. Colposcopy was performed on women with any degree of atypia on cytology and/or a positive high-risk human papillomavirus (HPV)-DNA test (HC2; Hybrid Capture 2((c))), and for a randomly selected sample of 3.4% women with negative findings on both. Quality control included reviews of cytology, histology, colposcopy images and retesting of samples with polymerase chain reaction. Test diagnostic performances were based on 7908 women who had complete baseline and follow-up results. Routine histology identified 86 women with high-grade cervical intraepithelial neoplasia (CIN2+), which was confirmed by review histology in only 46 cases. Sensitivity of routine cytology for the detection of CIN2+ was 43.5%, with a specificity, positive predictive value (PPV), negative predictive value (NPV) of 98.0, 11.4 and 99.7%, respectively. Sensitivity of the HC2 test for the detection of CIN2+ was 97.8%, with a specificity, PPV and NPV, of 95.3, 10.9 and 100%, respectively. No high-grade neoplasia was detected in the randomly selected control group. A negative HPV test result, even in combination with a positive Papanicolaou (Pap) result, virtually excluded any risk of underlying high-grade disease, but this was not the case for a negative Pap result. These data show that HPV testing is of value for the detection or exclusion of prevalent CIN in a routine cervical cancer screening setting and could be used for further risk classification of women for follow-up management. PMID- 12771925 TI - Cytogenetic alterations in ovarian clear cell carcinoma detected by comparative genomic hybridisation. AB - Ovarian clear cell carcinoma (OCCC) accounts for a small but significant proportion of all ovarian cancers and is a distinct clinical and pathological entity. It tends to be associated with poorer response rates to chemotherapy and with a worse prognosis. Little is known about possible underlying genetic changes. DNA extracted from paraffin-embedded samples of 18 pure OCCC cases was analysed for genetic imbalances using comparative genomic hybridisation (CGH). All of the 18 cases showed genomic alterations. The mean number of alterations detected by CGH was 6 (range 1-15) indicating a moderate level of genetic instability. Chromosome deletions were more common than amplifications. The most prominent change involved chromosome 9 deletions in 10 cases (55%). This correlates with changes seen in other epithelial ovarian cancers. This deletion was confirmed using microsatellite markers to assess loss of heterozygosity (LOH) at four separate loci on chromosome 9. The most distinct region of loss detected was around the IFNA marker at 9p21 with 41% (11 out of 27 cases) LOH. Other frequent deletions involved 1p (five out of 18; 28%); 11q (four out of 18; 22%) and 16 (five out of 18; 28%). Amplification was most common at chromosome 3 (six out of 18; 33%); 13q (four out of 18; 22%) and 15 (three out of 18; 17%). No high level amplifications were identified. These features may serve as useful prognostic indicators in the management of OCCC. PMID- 12771926 TI - Genetic alterations in the coding region of the bak gene in uterine cervical carcinoma. AB - A significant frequency of mutations (six missense and one silent) was found, for the first time, at the coding region of the bak gene (exons 3, 4 and 6) in 42 carcinomas of the uterine cervix, while no mutations were detected in 32 non neoplastic cervix tissues. Bak mutations were observed more frequently in the advanced stage and mutated cancer tissues were more resistant to radiotherapy, although trends were not statistically significant because of small sample size. PMID- 12771927 TI - Agreement between chromogenic in situ hybridisation (CISH) and FISH in the determination of HER2 status in breast cancer. AB - Determination of the HER2/neu (HER2) status in breast carcinoma has become necessary for the selection of breast cancer patients for trastuzumab therapy. Amplification of the gene analysed by fluorescence in situ hybridisation (FISH) or overexpression of the protein determined by immunohistochemistry (IHC) are the two major methods to establish this status. A strong correlation has been previously demonstrated between these two methods. However, FISH is not always feasible in routine practice and weakly positive IHC tumours (2+) do not always correspond to a gene amplification. Our study was performed in order to evaluate the contribution of chromogenic in situ hybridisation (CISH), which enables detection of the gene copies through an immunoperoxidase reaction. CISH was performed in 79 breast carcinomas for which the HER2 status was previously determined by IHC and FISH. The results of IHC, FISH and CISH were compared for each tumour. CISH procedures were successful in 95% of our cases. Whatever the IHC results, we found a very good concordance (96%) between CISH and FISH. Our study confirms that CISH may be an alternative to FISH for the determination of the gene amplification status in 2+ tumours. Our results allow us to think that, in many laboratories, CISH may also be an excellent method to calibrate the IHC procedures or, as a quality control test, to check regularly that the IHC signal is in agreement with the gene status. PMID- 12771928 TI - Tumour dose response to the antivascular agent ZD6126 assessed by magnetic resonance imaging. AB - ZD6126 is a vascular targeting agent that disrupts the tubulin cytoskeleton of proliferating neo-endothelial cells. This leads to the selective destruction and congestion of tumour blood vessels in experimental tumours, resulting in extensive haemorrhagic necrosis. In this study, the dose-dependent activity of ZD6126 in rat GH3 prolactinomas and murine RIF-1 fibrosarcomas was assessed using two magnetic resonance imaging (MRI) methods. Dynamic contrast-enhanced (DCE) MRI, quantified by an initial area under the time-concentration product curve (IAUC) method, gives values related to tumour perfusion and vascular permeability. Multigradient recalled echo MRI measures the transverse relaxation rate T(2)*, which is sensitive to tissue (deoxyhaemoglobin). Tumour IAUC and R(2)* (=1/T(2)*) decreased post-treatment with ZD6126 in a dose-dependent manner. In the rat model, lower doses of ZD6126 reduced the IAUC close to zero within restricted areas of the tumour, typically in the centre, while the highest dose reduced the IAUC to zero over the majority of the tumour. A decrease in both MRI end points was associated with the induction of massive central tumour necrosis measured histologically, which increased in a dose-dependent manner. Magnetic resonance imaging may be of value in evaluation of the acute clinical effects of ZD6126 in solid tumours. In particular, measurement of IAUC by DCE MRI should provide an unambiguous measure of biological activity of antivascular therapies for clinical trial. PMID- 12771929 TI - Upregulation of cyclooxygenase-2 is accompanied by increased expression of nuclear factor-kappa B and I kappa B kinase-alpha in human colorectal cancer epithelial cells. AB - Cyclooxygenase-2 (COX-2) is selectively overexpressed in colorectal tumours. The mechanism of COX-2 induction is not fully understood, but requires de novo messenger RNA and protein synthesis, indicating regulation at the transcriptional level. Sequence analysis of the 5'-flanking region of the COX-2 gene shows two nuclear factor-kappa B (NF-kappa B) sites. Inhibition of this protein in model cell culture systems attenuates COX-2 expression and implies that NF-kappa B plays an important role in COX-2 induction. We measured COX-2, NF-kappa B and I kappa B kinase alpha (IKK alpha) protein expression in matched colonic biopsy samples comprising both nontumour and adjacent tumour tissue from 32 colorectal cancer patients using immunohistochemistry. There was none or very little expression of COX-2, NF-kappa B and IKK alpha in non-neoplastic colon epithelial cells, while the expression of all three of these proteins was significantly increased (P<0.05, Wilcoxon's signed rank test) in adjacent cancerous cells. Moreover, all three proteins were found to be coexpressed in the neoplastic epithelium, with the expression of COX-2 and NF-kappa B highly correlated (Pearson's correlation, P<0.005). There was no apparent correlation between enhanced COX-2, NF-kappa B or IKK alpha expression and tumour Dukes' stages. Our results are compatible with the hypothesis that IKK alpha and NF-kappa B are involved in COX-2 induction in these tumours and the lack of association between COX-2 expression and severity of disease as measured by Dukes' stage is consistent with the proposal that COX-2 expression is an early postinitiation event. PMID- 12771930 TI - A novel IL-10 signalling mechanism regulates TIMP-1 expression in human prostate tumour cells. AB - We have previously reported that interleukin 10 (IL-10) signalling stimulated activation of a specific enhancer element, termed HTE-1, to promote tissue inhibitor of matrix metalloproteinase1 (TIMP-1) expression in human bone metastatic PC-3 subclone (PC-3 ML) cells. Recently, we have identified an IL-10 responsive signal molecule, termed IL-10E1, which binds the HTE-1 element and cloned the gene encoding for the 22 kDa protein. In this paper, we have examined the mechanism of IL-10/IL-10 receptor signalling in two distinct human prostate cell lines, a 'normal' prostate epithelial cell line, termed NPTX-1532 and highly metastatic PC-3 ML tumour cells. Signalling cascade studies revealed that IL-10 stimulated tyrosine phosphorylation of JAK1 and TYK2 receptor kinases and tyrosine phosphorylation of IL-10E1. Phosphorylation, triggered IL-10E1's rapid translocation to the nucleus by 10-30 min. Deletion analysis combined with transient transfection experiments revealed that the n-terminal domain (approximately 74 a.a.) of the IL-10E1 protein, the nt-nls peptide, was stimulated by IL-10 to translocate to the nucleus and induce TIMP-1 expression. Site-directed mutagenesis further showed that phosphorylation of two tyrosine moieties (Y57 and Y62) of the nt-nls peptide was required for IL-10 activation of signalling and TIMP-1 expression. The data demonstrate, for the first time, that IL-10 receptor signalling of TIMP-1 expression is regulated by tyrosine phosphorylation of a novel gene, IL-10E1, in human prostate cells. PMID- 12771931 TI - Quinazoline-based alpha 1-adrenoceptor antagonists induce prostate cancer cell apoptosis via TGF-beta signalling and I kappa B alpha induction. AB - Previous studies documented the ability of quinazoline-based alpha1-adrenoceptor antagonists to induce apoptosis in prostate cancer cells via an alpha 1 adrenoceptor-independent mechanism. In this study we investigated the molecular events initiating this apoptotic effect. Since transforming growth factor-beta 1 (TGF-beta 1) mediates prostate epithelial cell apoptosis, we hypothesised that the activation of the TGF-beta 1 pathway underlies the quinazoline-based apoptotic effect in prostate cancer cells. Treatment of the androgen-independent human prostate cancer cells PC-3 with doxazosin resulted in a strong caspase-3 activation within 24 h, whereas tamsulosin, a sulphonamide-based alpha 1 adrenoceptor antagonist, had no significant apoptotic effect against prostate cancer cells. To identify the molecular components involved in this quinazoline mediated apoptosis, cDNA microarray analysis of PC-3 prostate cancer cells treated with doxazosin (3 h) was performed. Induced expression of several genes was observed including p21(WAF-1) and I kappa B alpha (inhibitor of NF-kappa B alpha). Relative quantitative reverse transcription-polymerase chain reaction analysis revealed induction of several TGF-beta1 signalling effectors: Induction of mRNA for Smad4 and the TGF-beta1-regulated apoptosis-inducing transcription factor TGF-beta1-inducible early gene (TIEG1) was detected within the first 6 h of doxazosin treatment. Upregulation of I kappa B alpha at both the mRNA and protein level was also detected after 6 h of treatment. Furthermore, doxazosin resulted in a considerable elevation in Smad4 and TIEG protein expression (6 h). A 'latent' increase in TGF-beta mRNA expression was detected after 48 h of treatment. These findings suggest that the quinazoline-based doxazosin mediates prostate cancer apoptosis by initially inducing the expression of TGF-beta1 signalling effectors and subsequently I kappa B alpha. The present study provides an initial insight into the molecular targets of the apoptotic action of quinazolines against prostate cancer cells. PMID- 12771932 TI - A novel vascular endothelial growth factor-directed therapy that selectively activates cytotoxic prodrugs. AB - We have generated fusion proteins between vascular endothelial growth factor (VEGF) and the bacterial enzyme carboxypeptidase G2 (CPG2) that can activate the prodrug 4-[(2-chloroethyl)(2-mesyloxyethyl)amino]benzoyl-L-glutamic acid (CMDA). Three asparagine residues of CPG2 were mutated to glutamine (CPG2(Q)3) to prevent glycosylation during secretion, and truncations of VEGF(165) were fused to either the C- or N-terminal of CPG2. The K(m) of the fusion proteins (37.5 microM) was similar to that of secreted CPG2(Q)3 (29.5 microM) but greater than that of wild type CPG2 (8 microM). The affinity of the fusion proteins for VEGF receptor-2 (VEGFR2) (K(d)=0.5-1.1 nM) was similar to that of [(125)I]VEGF (K(d)=0.5 nM) (ELISA) or slightly higher (K(d)=1.3-9.6 nM) (competitive RIA). One protein, VEGF(115)-CPG2(Q)3-H(6), possessed 140% of the enzymic activity of secreted CPG2(Q)3, and had a faster half-maximal binding time for VEGFR2 (77 s), than the other candidates (330 s). In vitro, VEGF(115)-CPG2(Q)3-H(6) targeted CMDA cytotoxicity only towards VEGFR-expressing cells. The plasma half-life of VEGF(115)-CPG2(Q)3-H(6) in vivo was 3 h, comparable to equivalent values observed in ADEPT. We conclude that enzyme prodrug therapy using VEGF as a targeting moiety represents a promising novel antitumour therapy, with VEGF(115)-CPG2(Q)3 H(6) being a lead candidate. PMID- 12771933 TI - New insights into the actions of bisphosphonate zoledronic acid in breast cancer cells by dual RhoA-dependent and -independent effects. AB - Zoledronic acid (ZOL) is a nitrogen-containing bisphosphonate and its use in reducing osteoporosis and cancer-induced osteolysis is increasing. Recent findings indicated that ZOL has a direct effect on cancer cells. In this study, the effect of ZOL was examined on the aggressive MDA-MB-231 breast cancer cell line. ZOL induces an important inhibition of cell invasion at low concentrations (1 microM). This is not explained by modifications of proteases involved in cell invasiveness (matrix metalloproteinases and urokinase-type plasminogen activator), but by a disorganisation of actin cytoskeleton due to RhoA inhibition related to its defective prenylation as it was reversed by geranylgeraniol (GGOH) and mimicked by the Rho selective inhibitor C3 exoenzyme. In addition, ZOL inhibits the chemotactic effect induced by stromal cell-derived factor 1(SDF-1), a chemokine greatly involved in cancer metastasis to bone. This effect is related to both reduction of cell motility induced by RhoA inhibition and to a decreased expression of CXCR-4, the SDF-1 receptor. Finally, ZOL reduces Cox-2 expression and, consequently, the secretion of prostaglandins E2 (PGE2) in a RhoA independent manner. This inhibition could contribute to bone protection in breast cancers because PGE2 stimulates osteoclast-mediated bone resorption. In summary, new insights in the mechanism of ZOL action on aggressive breast cancer cells are demonstrated and could explain its beneficial action in both the reduction of osteolysis and prevention of metastasis. PMID- 12771934 TI - Improved antitumour immunity in murine neuroblastoma using a combination of IL-2 and IL-12. AB - Neuroblastoma immunotherapy using cytokine-modified tumour cells has been tested in clinical trials. However, because of the complex nature of antitumour immune responses, a number of therapies may be required for complete tumour eradication and generation of systemic immunity. We report here the improved antitumour effect of two cytokines, interleukin-2 (IL-2) and interleukin-12 (IL-12), when coexpressed by neuroblastoma cell lines. Initially, transfection of human and mouse neuroblastoma cell lines resulted in high expression levels of biologically active IL-2 and IL-12 in vitro. These cytokines when expressed by transfected Neuro-2A cells completely abolished their in vivo tumorigenicity in a syngeneic neuroblastoma model. Vaccination of established tumours with IL-12-producing cells exhibited a clear effect with reduced tumour growth in the presence of IL 2. In vivo depletion studies showed that CD4(+) and CD8(+) T cells mediate the response against cytokine-producing cells. These results suggest that IL-2 and IL 12, when cotransfected in tumour cells, are effective against established disease and provide a promising immunotherapeutic approach for the treatment of neuroblastoma. PMID- 12771935 TI - Delayed expression of apoptosis in human lymphoma cells undergoing low-dose taxol induced mitotic stress. AB - The links between low-dose range taxol-induced mitotic arrest and the subsequent engagement of apoptosis are important for identifying the routes to therapeutic action. Here we have investigated the timing of cell-cycle perturbation and cell death responses following continuous exposure to clinically relevant drug concentrations (1-20 nM). Following 8 h of exposure to taxol, the cell line DoHH2 (p53 wild type) exhibited mitotic arrest and engagement of apoptosis, whereas the cell line SU-DHL-4 (p53 mutant) breached cell-cycle arrest with progression to an abnormal cycle and a 24 h delay in the engagement of apoptosis. Imaging showed equivalent dysfunction of mitotic spindles in both cell lines. The results of kinetic analyses indicated that although cell death may occur at different stages of progression through mitosis and subsequent cell cycles, the overall kinetics of cell death relate to the rate of arrival at a critical event window in the cell cycle. We propose a simple model of low-dose taxol-induced cell death for cycling populations in which mitotic stress acts as a primary trigger for apoptosis with equivalent but potentially delayed outcomes. This view provides a rationale for the clinical effectiveness of this agent, independent of the initial capacity of the tumour cell to engage apoptosis due, for example, to mutant p53 expression. The results provide a perspective for the design of combination regimens that include low-dose taxol and a component that may disturb mitotic delivery. PMID- 12771937 TI - Nitric oxide induces phosphorylation of p53 and impairs nuclear export. AB - The tumor suppressor p53 accumulates under diverse stress conditions and affects cell cycle progression and/or apoptosis. This has been exemplified for endogenously produced or exogenously supplied nitric oxide (NO) and thus accounts at least in part for pathophysiological signaling of that bioactive molecule, although detailed mechanisms remain to be elucidated. By using luciferase reporter assays, we show that NO stabilized a transcriptionally active p53 protein. Considering that p53 is targeted by murine double minute (Mdm2) for ubiquitination and subsequent proteasomal degradation and knowing that this interaction is impaired by, for example, UV-treatment with concomitant stabilization of p53 we questioned the p53/Mdm2 interaction in the presence of NO. Although p53 became phosphorylated at serine 15 under the impact of NO, coimmunoprecipitation with Mdm2 and ubiquitination remained intact, thus excluding any interference of NO with this pathway. The importance of N-terminal p53 phosphorylation was verified with p53 mutants where the first six serine residues have been converted to alanine, and which do not accumulate in response to NO. Regulation of p53 stability can be also achieved by affecting nuclear cytoplasmic shuttling and it was presented that leptomycin B, an inhibitor of nuclear export, caused p53 accumulation. Cell fractionation and immunofluorescence staining following NO-treatment revealed predominant nuclear accumulation of p53 in close association with serine 15-phosphorylation, which suggests impaired nuclear-cytoplasmic shuttling. This was verified by heterokaryon analysis. We conclude that attenuated nuclear export contributes to stabilization and activation of p53 under the influence of NO. PMID- 12771938 TI - Activation of ERK1/2 protects melanoma cells from TRAIL-induced apoptosis by inhibiting Smac/DIABLO release from mitochondria. AB - We have previously shown that Smac/DIABLO release from mitochondria appears to be the principal pathway by which TRAIL induces apoptosis of human melanoma. We report that TRAIL-induced release of Smac/DIABLO appears to be downregulated by concomitant signaling through the MEK Erk1/2 kinase pathway and that this inhibits TRAIL-induced apoptosis. Inhibition of Erk1/2 signaling by either the MEK inhibitor U0126 or a dominant-negative mutant of MKK1 markedly sensitized melanoma cells to TRAIL-induced apoptosis. The site in the apoptotic pathway acted on by U0126 appeared to be downstream of caspase-8 and Bid but upstream of caspase-3 in that the levels of proteolytic cleavage of caspase-8 and Bid by TRAIL were similar in cells with or without exposure to U0126. Caspase-3 activation and cleavage of its substrates, PARP, ICAD and XIAP, were however increased by cotreatment with U0126. This was associated with a rapid reduction in mitochondrial transmembrane potential (MMP) and increased release of Smac/DIABLO into the cytosol. Exploration of events leading to the changes in MMP revealed an increased translocation of Bax from the cytosol to mitochondria in the presence of U0126. There was also a delayed decrease in the levels of expression of Mcl-1. Bcl-2 and Bcl-X(L). Over expression of Bcl-2 blocked TRAIL induced apoptosis in the presence of U0126. Cytochrome c appeared not to play a major role in sensitization of melanoma to TRAIL in that caspase-9 activation was not detected in most of the cell lines. These results suggest that Erk1/2 signaling may protect melanoma cells against TRAIL-induced apoptosis by inhibiting the relocation of Bax from the cytosol to mitochondria and that this may reduce TRAIL-mediated release of Smac/DIABLO and induction of apoptosis. PMID- 12771939 TI - Identification and characterization of a PU.1/Spi-B binding site in the bovine leukemia virus long terminal repeat. AB - Bovine leukemia virus (BLV) is a B-lymphotropic oncogenic retrovirus whose transcriptional promoter is located in the viral 5' long terminal repeat (LTR). To date, no B-lymphocyte-specific cis-regulatory element has been identified in this region. Since ETS proteins are known to regulate transcription of numerous retroviruses, we searched for the presence in the BLV promoter region of binding sites for PU.1/Spi-1, a B-cell- and macrophage-specific ETS family member. In this report, nucleotide sequence analysis of the viral LTR identified a PUbox located at -95/-84 bp. We demonstrated by gel shift and supershift assays that PU.1 and the related Ets transcription factor Spi-B interacted specifically with this PUbox. A 2-bp mutation (GGAA-->CCAA) within this motif abrogated PU.1/Spi-B binding. This mutation caused a marked decrease in LTR-driven basal gene expression in transient transfection assays of B-lymphoid cell lines, but did not impair the responsiveness of the BLV promoter to the virus-encoded transactivator Tax(BLV). Moreover, ectopically expressed PU.1 and Spi-B proteins transactivated the BLV promoter in a PUbox-dependent manner. Taken together, our results provide the first demonstration of regulation of the BLV promoter by two B-cell-specific Ets transcription factors, PU.1 and Spi-B. The PU.1/Spi-B binding site identified here could play an important role in BLV replication and B-lymphoid tropism. PMID- 12771940 TI - ARHI is a Ras-related small G-protein with a novel N-terminal extension that inhibits growth of ovarian and breast cancers. AB - Our group recently identified Ras homolog member I (ARHI), a novel maternally imprinted tumor suppressor gene that encodes a 26 kDa GTP-binding protein with high homology to Ras and Rap. Unlike other Ras family members, ARHI exhibits several unusual structural and functional properties. ARHI contains a unique 34 amino-acid extension at the N-terminus, and differs from Ras in residues critical for GTPase activity and in its putative effector domain. Like Ras, ARHI can bind to GTP with high affinity but has low intrinsic GTPase activity. In addition, while Ras is an oncogene, ARHI functions as an inhibitor for cell growth. (32)Phosphorus labeling showed that ARHI is maintained in a constitutively activated GTP-bound state in resting cells, possibly because of impaired GTPase activity. ARHI is associated at the cell membrane through its prenylation at the C-terminal cysteine residue. Mutation of the conserved CAAX box at the C-terminus led to a loss of its membrane association and a decreased ability to inhibit cell growth. Conversion of Ser(51) to Asn decreased GTP binding and reduced ARHI's biological activity. Mutation of Ala(46) to Val increased the ability of ARHI to inhibit cell growth, associated with a further decrease of its intrinsic GTPase activity. Moreover, conversion of residues in ARHI that are conserved in the Ras family for GTPase activity partially restored the GTPase activity in ARHI. Most strikingly, deletion of ARHI's unique N-terminal extension nearly abolished its inhibitory effect on cell growth, suggesting its importance in ARHI's inhibitory function. Thus, ARHI is a unique Ras family member that retains basic small GTPase function, but exhibits many unusual features. In contrast to most other Ras family members, ARHI has a long N-terminal extension, modest GTPase activity, and constitutive GTP binding in resting cells. Furthermore, unlike the Ras oncogene, ARHI inhibits cell growth, and loss of its expression in cells may contribute to the development of breast and ovarian cancers. PMID- 12771941 TI - HBX causes cyclin D1 overexpression and development of breast cancer in transgenic animals that are heterozygous for p53. AB - Transgenic mice, which selectively express the WAP-HBX transgene in mammary gland epithelial cells (ME-cells), were established in order to elucidate the consequences of HBX gene expression on organ differentiation, cell death program and tumor development. Transgene expression was demonstrable by RT-PCR, Northern and Western blot analysis during pregnancy, lactation and after weaning. HBX synthesis neither affect mammary gland differentiation nor apoptosis in ME-cells. Although breast cancer formation was rare in WAP-HBX animals (<1%), WAP-HBX*p53+/ hybrid animals developed breast tumors at an increased rate (12/85) after a latency period of 8-18 months. We also show here for the first time that HBX can immortalize ME-cells generated from mammary gland tissue segments in a p53 independent fashion. HBX causes cyclin D1 gene overexpression during early pregnancy, and this is maintained in ME-cells isolated either from mammary gland or from breast tumors. Intranuclear cyclin D1 accumulation also occurs in the absence of external growth factors and the BrdU incorporation rate remains high under serum starvation conditions. Finally, both cyclin D1 induction and HBX mitotic activity are dependent on p38 and c-Jun N-terminal kinase, but not on MEK 1 kinase activity. PMID- 12771942 TI - REPS2/POB1 is downregulated during human prostate cancer progression and inhibits growth factor signalling in prostate cancer cells. AB - During progression of prostate cancer, cellular changes occur, leading to a transition from androgen-dependent to androgen-independent growth. One aspect of this transition is a switch from androgens to growth factors, like epidermal growth factor (EGF), as primary regulators of proliferation. We examined the involvement of REPS2/POB1 in this process. REPS2/POB1 is an EH domain-containing protein, reported to be involved in signalling via RalBP1 and to play a role in endocytosis of EGF receptors. Furthermore, the protein is relatively highly expressed in androgen-dependent as compared to androgen-independent human prostate cancer cell lines and xenografts. Next to the known REPS2/POB1 protein, an open reading frame encoding REPS2/POB1, with 139 additional amino-acid residues at the NH(2)-terminus, was cloned and found to be expressed in prostate cancer cells. Overexpression, by transient transfection, of both forms of REPS2/POB1 in prostate cancer cell lines, induced apoptosis within 48 h. At shorter time intervals after transfection, signalling towards a TPA response element luciferase reporter was found to be inhibited. From these experiments, it is concluded that REPS2/POB1, through its influence on the Ral signalling pathway, is involved in growth factor signalling. Decreased expression of REPS2/POB1 during progression of prostate cancer may therefore result in loss of control of growth factor signalling and consequently in loss of control of cell proliferation. PMID- 12771943 TI - Cell cycle and genetic background dependence of the effect of loss of BRCA2 on ionizing radiation sensitivity. AB - Carriers of mutations in the BRCA2 gene are at a highly elevated risk of breast and other cancers. The BRCA2 gene encodes a very large protein thought to play a role in DNA repair. To examine the effect of mutation of BRCA2 on sensitivity to ionizing radiation, we used a previously described mouse model system (Brca2(Tr)) in which the Brca2 open reading frame is truncated. Mouse embryo fibroblasts carrying this mutation have a proliferative defect, which we show here can be substantially rescued by genetic ablation of p53. Proliferating Brca2(Tr/Tr)/p53( /-) cells, like Brca2(Tr/Tr) cells, show genomic instability. We used the clonogenic survival assay, which depends on the ability of cells to proliferate, to examine the cell cycle dependence of radiation sensitivity of Brca2(Tr/Tr)/p53(-/-) compared to p53(-/-) and wild-type cells. This showed that the Brca2 mutation had little effect on cells irradiated in quiescence but sensitized proliferating cells to ionizing radiation on a p53(-/-) background. These results suggest that the major role of Brca2 in mediating cell survival after irradiation is in the S and G(2) phases of the cell cycle. PMID- 12771944 TI - Transactivation mediated by B-Myb is dependent on TAF(II)250. AB - B-Myb is a highly conserved member of the Myb family of transcription factors, which has been implicated in cell cycle regulation. B-Myb is expressed in most proliferating cells and its activity is highly regulated around the G1/S-phase border of the cell cycle. It is generally assumed that B-Myb regulates the expression of genes that are crucial for cell proliferation; however, the identity of these genes, the molecular mechanisms by which B-Myb stimulates their expression and the involvement of other proteins have not been sufficiently clarified. We have employed the hamster cell line ts13 as a tool to demonstrate a functional link between B-Myb and the coactivator TAF(II)250, a key component of the transcriptional machinery which itself is essential for cell proliferation. ts13 cells express a point-mutated version of TAF(II)250 whose intrinsic histone acetyl transferase activity is temperature sensitive. Transactivation of Myb responsive reporter genes by B-Myb is temperature-dependent in ts13 cells but not in ts13 cells, which have been rescued by transfection with an expression vector for wild-type TAF(II)250. Furthermore, B-Myb and TAF(II)250 can be coprecipitated, suggesting that both proteins are present in a complex. The formation of this complex is dependent on the DNA-binding domain of B-Myb and not on its transactivation domain. Taken together, these observations provide the first evidence that the coactivator TAF(II)250 is involved in the activation of Myb responsive promoters by B-Myb. The finding that B-Myb transactivation is dependent on a key coactivator involved in cell cycle control is consistent with and strengthens the idea that B-Myb plays a crucial role as a transcription factor in proliferating cells. PMID- 12771945 TI - Protein kinase Calpha activation by RET: evidence for a negative feedback mechanism controlling RET tyrosine kinase. AB - We have studied the role of protein kinase C (PKC) in signaling of the RET tyrosine kinase receptor. By using a chimeric receptor (E/R) in which RET kinase can be tightly controlled by the addition of epidermal growth factor (EGF), we have found that RET triggering induces a strong increase of PKCalpha, PKCdelta and PKCzeta activity and that PKCalpha, not PKCdelta and PKCzeta, forms a ligand dependent protein complex with E/R. We have identified tyrosine 1062 in the RET carboxyl-terminal tail as the docking site for PKCalpha. Block of PKC activity by bisindolylmaleimide or chronic phorbol esters treatment decreased EGF-induced serine/threonine phosphorylation of E/R, while it caused a similarly sized increase of EGF-induced E/R tyrosine kinase activity and mitogenic signaling. Conversely, acute phorbol esters treatment, which promotes PKC activity, increased the levels of E/R serine/threonine phosphorylation and significantly decreased its phosphotyrosine content. A threefold reduction of tyrosine phosphorylation levels of the constitutively active RET/MEN2A oncoprotein was observed upon coexpression with PKCalpha. We conclude that RET binds to and activates PKCalpha. PKCalpha, in turn, causes RET phosphorylation and downregulates RET tyrosine kinase and downstream signaling, thus functioning as a negative feedback loop to modulate RET activity. PMID- 12771947 TI - Resistance to DNA-damaging agents is discordant from experimental metastatic capacity in MEF ras-transformants-expressing gain of function MTp53. AB - Tumor cells can acquire aggressive phenotypes secondary to the loss of expression of the wild-type p53 (WTp53) protein or by the gain of function for selected mutant p53 (MTp53) proteins. However, it is unclear as to whether the development of aggressive phenotypes is inter-related. Herein we report the radiosensitivity, chemosensitivity, and in vivo growth characteristics of isogenic p53(-/-) MEF ras transformants that variably express an MTp53 protein. Initial experiments revealed significant clonal heterogeneity with respect to cellular sensitivity to DNA-damaging agents (i.e. ionizing radiation, ultraviolet radiation, cis platinum, and methotrexate) within subclones of a pre-existing p53(-/-) MEF cell population. Moreover, this differential sensitivity was also observed within subclones of p53(-/-) MEF cells transformed with an activated ras allele, suggesting that secondary genetic events and clonal selection, but not cellular transformation per se, may drive the resistance patterns for certain null-p53 tumors. In contrast, uniform resistance was observed following the additional transfection of an MTp53 allele (MTp53pro193) into p53(-/-) MEF transformants and p53(-/-) DP-16 Friend erythroleukemia cells, consistent with a gain of MTp53 function for this allele. Relative tumor growth rate and experimental metastatic ability was not enhanced by MTp53pro193 expression. Our results support the concept that gain of MTp53pro193 function leads to the selection of dominant clones, which may exhibit cellular resistance following cancer therapy. PMID- 12771946 TI - A major role for Mcl-1 antiapoptotic protein in the IL-6-induced survival of human myeloma cells. AB - Interleukin-6 (IL-6) is a major survival factor for malignant plasma cells involved in multiple myeloma. Using an RNase protection assay, we looked for gene expression of 10 anti- and proapoptotic Bcl-2-family proteins in 12 IL-6 dependent human myeloma cell lines (HMCL). A high Mcl-1 gene expression was found in all HMCLs and the other genes were variably expressed. Out of the 10 Bcl-2 family members, only the Mcl-1 gene was regulated by IL-6. Upon starvation of IL 6, Mcl-1 gene expression decreased in association with myeloma cell apoptosis and was upregulated after adding IL-6 again in association with myeloma cell survival. A constitutive Mcl-1 expression was induced with an Mcl-1-GFP retrovirus in two IL-6-dependent HMCLs. The Mcl-1 HMCLs have a marked reduced apoptosis upon IL-6 starvation compared to HMCLs transduced with control GFP retrovirus and may grow without adding IL-6. These data emphasize the major role of Mcl-1 antiapoptotic protein in the IL-6-induced survival of human myeloma cells. PMID- 12771949 TI - SASH1: a candidate tumor suppressor gene on chromosome 6q24.3 is downregulated in breast cancer. AB - Loss of heterozygosity (LOH) and in silico expression analysis were applied to identify genes significantly downregulated in breast cancer within the genomic interval 6q23-25. Systematic comparison of candidate EST sequences with genomic sequences from this interval revealed the genomic structure of a potential target gene on 6q24.3, which we called SAM and SH3 domain containing 1 (SASH1). Loss of the gene-internal marker D6S311, found in 30% of primary breast cancer, was significantly correlated with poor survival and increase in tumor size. Two SASH1 transcripts of approximately 4.4 and 7.5 kb exist and are predominantly transcribed in the human breast, lung, thyroid, spleen, placenta and thymus. In breast cancer cell lines, SASH1 is only expressed at low levels. SASH1 is downregulated in the majority (74%) of breast tumors in comparison with corresponding normal breast epithelial tissues. In addition, SASH1 is also downregulated in tumors of the lung and thyroid. Analysis of the protein domain structure revealed that SASH1 is a member of a recently described family of SH3/SAM adapter molecules and thus suggests a role in signaling pathways. We assume that SASH1 is a new tumor suppressor gene possibly involved in tumorigenesis of breast and other solid cancers. We were unable to find mutations in the coding region of the gene in primary breast cancers showing LOH within the critical region. We therefore hypothesize that other mechanisms as for instance methylation of the promoter region of SASH1 are responsible for the loss of expression of SASH1 in primary and metastatic breast cancer. PMID- 12771948 TI - Alteration of the PATCHED locus in superficial bladder cancer. AB - Chromosome 9 alterations are the most frequently encountered cytologic anomalies in urothelial carcinoma (UC). We previously screened 139 low-stage UCs for loss of heterozygosity on chromosome 9, and identified five distinct regions likely to harbour tumour-suppressor genes. The present study focused on deletion mapping in the 9q22 region with 11 additional microsatellite markers. New deletions in the 9q22 region were found in five tumours. Deletion mapping allowed us to identify a 0.5 CM common minimal region of deletion between markers D9S280 and D9S1809, encompassing PATCHED (PTC), a gene identified as a tumour suppressor in basal cell carcinoma and in medulloblastoma. A marker located in the first intron of this gene showed the highest percentage of deletion (45%). cDNA sequencing in 15 tumours with deletion of PTC showed no mutation in the remaining allele. However, average expression of PTC mRNA measured by semiquantitative RT-PCR was significantly decreased in tumours with LOH in the 9q22 region, compared to normal urothelium (P=0.04), while it showed marked fluctuations in tumours without deletion. Our results suggest that the PTC gene is a putative suppressor at the 9q22 locus and that haploinsufficiency of this gene may be an early event in the development of papillary bladder tumours. PMID- 12771950 TI - Deletion mapping using quantitative real-time PCR identifies two distinct 3p21.3 regions affected in most cervical carcinomas. AB - We report chromosome 3p deletion mapping of 32 cervical carcinoma (CC) biopsies using 26 microsatellite markers located in frequently deleted 3p regions to detect loss of heterozygosity and homozygous loss. In addition, two STS markers (NLJ-003 and NL3-001) located in the 3p21.3 telomeric (3p21.3T) and 3p21.3 centromeric (3p21.3C) regions, respectively, were used for quantitative real-time PCR as TaqMan probes. We show that quantitative real-time PCR is reliable and sensitive and allows discriminating between 0, 1 and 2 marker copies per human genome. For the first time, frequent (five of 32 cases, i.e. 15.6%) homozygous deletions were demonstrated in CCs in both 3p21.3T and 3p21.3C regions. The smallest region homozygously deleted in 3p21.3C was located between D3S1568 (CACNA2D2 gene) and D3S4604 (SEMA3F gene) and contains 17 genes previously defined as lung cancer candidate Tumor suppressor genes (TSG(s)). The smallest region homozygously deleted in 3p21.3T was flanked by D3S1298 and NL1-024 (D3S4285), excluding DLEC1 and MYD88 as candidate TSGs involved in cervical carcinogenesis. Overall, this region contains five potential candidates, namely GOLGA4, APRG1, ITGA9, HYA22 and VILL, which need to be analysed. The data showed that aberrations of either NLJ-003 or NL3-001 were detected in 29 cases (90.6%) and most likely have a synergistic effect (P<0.01). The study also demonstrated that aberrations in 3p21.3 were complex and in addition to deletions, may involve gene amplification as well. The results strongly suggest that 3p21.3T and 3p21.3C regions harbor genes involved in the origin and/or development of CCs and imply that those genes might be multiple TSG(s). PMID- 12771951 TI - Genome-wide comparison of human keratinocyte and squamous cell carcinoma responses to UVB irradiation: implications for skin and epithelial cancer. AB - To gain insight into the transformation of epidermal cells into squamous carcinoma cells (SCC), we compared the response to ultraviolet B radiation (UVB) of normal human epidermal keratinocytes (NHEK) versus their transformed counterpart, SCC, using biological and molecular profiling. DNA microarray analyses (Affymetrix), approximately 12000 genes) indicated that the major group of upregulated genes in keratinocytes fall into three categories: (i). antiapoptotic and cell survival factors, including chemokines of the CXC/CC subfamilies (e.g. IL-8, GRO-1, -2, -3, SCYA20), growth factors (e.g. HB-EGF, CTGF, INSL-4), and proinflammatory mediators (e.g. COX-2, S100A9), (ii). DNA repair-related genes (e.g. GADD45, ERCC, BTG-1, Histones), and (iii). ECM proteases (MMP-1, -10). The major downregulated genes are DeltaNp63 and PUMILIO, two potential markers for the maintenance of keratinocyte stem cells. NHEK were found to be more resistant than SCC to UVB-induced apoptosis and this resistance was mainly because of the protection from cell death by secreted survival factors, since it can be transferred from NHEK to SCC cultures by the conditioned medium. Whereas the response of keratinocytes to UVB involved regulation of key checkpoint genes (p53, MDM2, p21(Cip1), DeltaNp63), as well as antiapoptotic and DNA repair-related genes - no or little regulation of these genes was observed in SCC. The effect of UVB on NHEK and SCC resulted in upregulation of 251 and 127 genes, respectively, and downregulation of 322 genes in NHEK and 117 genes in SCC. To further analyse these changes, we used a novel unsupervised coupled two way clustering method that allowed the identification of groups of genes that clearly partitioned keratinocytes from SCC, including a group of genes whose constitutive expression levels were similar before UVB. This allowed the identification of discriminating genes not otherwise revealed by simple static comparison in the absence of UVB irradiation. The implication of the changes in gene profile in keratinocytes for epithelial cancer is discussed. PMID- 12771952 TI - Differential gene expression in distinct virologic types of hepatocellular carcinoma: association with liver cirrhosis. AB - Using oligonucleotide microarray data of 45 hepatocellular carcinoma (HCC) samples, we evaluated gene expression in hepatitis B virus-positive and hepatitis C virus-positive HCCs (HBV- and HCV-HCCs) for an association with liver cirrhosis (LC). In all, 89 genes were expressed differentially between HBV-HCCs associated with LC and those not associated with LC. Among them, tumors from LC patients showed significantly lower expression levels of 72 genes and significantly higher levels of 17 genes than the levels found in tumors from non-LC patients. The former included genes responsible for signal transduction, transcription, metabolism, and cell growth. The latter included a tumor suppressor gene and a cell-growth-related gene. Only eight genes were expressed differentially between HCV-HCCs associated with and without LC. Our findings provide as a framework for clarifying the role of LC in HBV- and HCV-related hepatocarcinogenesis. PMID- 12771955 TI - Basic neuroscience research has important implications for child development. PMID- 12771956 TI - Synchrony is stubborn in feedforward cortical networks. PMID- 12771957 TI - A signature of salience in the Drosophila brain. PMID- 12771958 TI - Aging, spatial behavior and the cognitive map. PMID- 12771959 TI - Motion perception: read my LIP. PMID- 12771962 TI - Rheb is a direct target of the tuberous sclerosis tumour suppressor proteins. AB - Mutations in the TSC1 or TSC2 genes cause tuberous sclerosis, a benign tumour syndrome in humans. Tsc2 possesses a domain that shares homology with the GTPase activating protein (GAP) domain of Rap1-GAP, suggesting that a GTPase might be the physiological target of Tsc2. Here we show that the small GTPase Rheb (Ras homologue enriched in brain) is a direct target of Tsc2 GAP activity both in vivo and in vitro. Point mutations in the GAP domain of Tsc2 disrupted its ability to regulate Rheb without affecting the ability of Tsc2 to form a complex with Tsc1. Our studies identify Rheb as a molecular target of the TSC tumour suppressors. PMID- 12771963 TI - Validation of a semi-quantitative adolescent food frequency questionnaire applied at a public school in Sao Paulo, Brazil. AB - OBJECTIVE: To develop a food frequency questionnaire for adolescents (AFFQ) and demonstrate its relative validity. DESIGN: The final version of the AFFQ was composed of 76 food items previously identified according to their contribution in nutrients and overall importance within the eating habits of this population group. The validation study, which was undertaken during a 6 month period (June to November 1999), was administered to a sample of 79 who answered at least three 24 h dietary recalls (R24 h) applied at intervals of 45 days and one AFFQ at the end of the study. Applying the paired t-test and calculating Pearson correlation coefficients on nutrient data, differences in the mean of nutrients were obtained. Correlation coefficients between the mean energy-adjusted nutrients computed by the two methods were calculated, and correction was made for within person variability. Agreement was evaluated by distribution of the adolescents according to quartiles of consumption. LOCUS: A public school within the metropolitan region of Sao Paulo city. RESULTS: A high variability in the dietary intake of adolescents was observed, with high rates of variability for cholesterol, retinal and vitamin C. The Pearson correlation coefficients, after being adjusted and corrected for variability, ranged from 0.10 to 0.72 among females and from 0.16 and 0.91 among males. The mean correlation coefficient for the entire group was 0.52. CONCLUSIONS: These results indicate that the AFFQ provides a potentially reliable scale for categorizing individuals by level of past intake of most nutrients, excluding retinol and iron. PMID- 12771964 TI - Swiss pilot study of individual seasonal fluctuations of urinary iodine concentration over two years: is age-dependency linked to the major source of dietary iodine? AB - OBJECTIVE: Because of known significant seasonal fluctuations of iodine concentration in cow's milk (ICM) in Switzerland (winter/summer (w/s) ICM ratio averages 5.6), we looked for seasonal variations and familial aspects in urinary iodine and sodium concentrations (UIC, UNaC). DESIGN: : Prospective sequential cohort investigation. SETTING: University hospital. SUBJECTS: Thirteen children (six girls, seven boys) aged 3-10 y and their parents (n=9) aged 30-47 y. INTERVENTIONS: The volunteers collected 1729 spot urine samples (5.1996-5.1998). UIC data from winter (UIC(w): December-February) and summer seasons (UIC(s): July September) were compared with UNaC and average milk consumption. Iodine intakes from milk and salt were modelized. RESULTS: Highest seasonal fluctuations of UIC were found in six pre-school children (w/s UIC group average=1.56, significant), followed by seven school children (w/s UIC group average=1.24, N.S.); none existed in adults. UIC/UNaC showed corresponding seasonal fluctuations in children, but not in adults. Winter milk was an important iodine source for children, as proven indirectly by similar seasonal fluctuations of ICM, UIC, UIC/UNaC and an important part within UIC due to milk. CONCLUSIONS: Contribution to UIC from milk intake during winter was high in children (40-50%) and lower in adults (about 20%). Compared with children, dietary habits of adults are more complex and their iodine supply depends mainly on iodized salt and not on milk, so the effect of seasonal ICM variations on UIC is less marked. Because of significant seasonal UIC fluctuations in consumers of fresh milk products in Switzerland, results of future UIC studies conducted during summer and winter seasons should be compared cautiously, especially in young children. Furthermore, consumption of milk is to be promoted, since, besides calcium and vitamins, it is an essential source of iodine. UNaC determination should be included in epidemiologic studies if the dietary source of iodine is questioned. PMID- 12771965 TI - Total energy expenditure and physical activity level in healthy young Swedish children 9 or 14 months of age. AB - OBJECTIVES: To measure total energy expenditure (TEE) and total body water (TBW) in healthy Swedish children 9 or 14 months of age. To compare their TEE with current recommendations for energy intake. To define their body composition and relate this to energy expenditure. DESIGN: : Children were investigated at 9 or 14 months. The following variables were measured: TEE and TBW (by the doubly labelled water method), weight and length. Total body fat (TBF), sleeping metabolic rate, activity energy expenditure and physical activity level (PAL) were calculated. SUBJECTS: Thirty infants 9 months of age and 29 children 14 months of age. RESULTS: : TEE was 323+/-38, 322+/-29, 313+/-23 and 331+/-28 kJ/kg/day in 9-month-old girls, 9-month-old boys, 14-month-old girls and 14-month old boys, respectively. At 9 months of age girls and boys contained 29.6+/-4.8 and 29.7+/-4.5% TBF, respectively. At 14 months the corresponding figures were 29.1+/-4.3 and 28.2+/-4.3%. There was a significant negative relationship between PAL and %TBF (r=-0.81, P<0.001, n=59). CONCLUSIONS: Measured TEE plus calculated energy cost of growth confirm previous estimates that the physiological energy requirements of children 9 and 14 months of age are 15-20% lower than current recommendations for energy intake. One possible interpretation of the relationship between PAL and %TBF is that children with a high TBF content are less physically active than children with less TBF. However, this relationship needs further studies. PMID- 12771967 TI - Evaluation of a dietary targets monitor. AB - OBJECTIVE: To evaluate a two-page food frequency list for use as a Dietary Targets Monitor in large scale surveys to quantify consumptions of the key foods groups targeted in health promotion. DESIGN: Intakes of fruit and vegetables, starchy foods and fish estimated from a validated food frequency questionnaire (FFQ) were compared with a short food frequency list (the Dietary Targets Monitor) specifically designed to assess habitual frequency of consumption of foods in relation to dietary targets which form the basis of a National (Scottish) Food and Health Policy. SUBJECTS: A total of 1085 adults aged 25-64 y from the Glasgow MONICA Study. RESULTS: : The two questionnaires both collected data on frequencies of food consumption for fruit and vegetables, starchy foods and fish. Comparing the two questionnaires, there were consistent biases, best expressed as ratios (FFQ:Dietary Targets Monitor) between the methods for fruit and vegetables (1.33, 95% CI 1.29, 1.38) and 'starchy foods' (1.08, 95% CI 1.05, 1.12), the DTM showing systematic under-reporting by men. For fish consumption, there was essentially no bias between the methods (0.99, 95% CI 0.94, 1.03). Using calibration factors to adjust for biases, the Dietary Targets Monitor indicated that 16% of the subjects were achieving the Scottish Diet food target (400 g/day) for fruit and vegetable consumption. Nearly one-third (32%) of the subjects were eating the recommended intakes of fish (three portions per week). The Dietary Targets Monitor measure of starchy foods consumption was calibrated using FFQ data to be able to make quantitative estimates: 20% of subjects were eating six or more portions of starchy food daily. A similar estimation of total fat intake and saturated fat intake (g/day) allowed the categorization of subjects as low, moderate or high fat consumers, with broad agreement between the methods. The levels of agreement demonstrated by Bland-Altman analysis, were insufficient to permit use of the adjusted DTM to estimate quantitative consumption in smaller subgroups. CONCLUSIONS: The Dietary Targets Monitor provides a short, easily administered, dietary assessment tool with the capacity to monitor intakes for changes towards national dietary targets for several key foods and nutrients. PMID- 12771966 TI - Nutrient intake and iron status of urban poor and rural poor without access to rice fields are affected by the emerging economic crisis: the case of pregnant Indonesian women. AB - OBJECTIVE: We evaluated the adequacy of nutrient intake in comparison with the Indonesian Estimated Average Requirement (EARs) among pregnant Indonesian women and explain the short-term effect of economic crisis on nutrient intake and iron status. DESIGN: Cross-sectional study. SETTING: Purworejo District, Central Java, located 60 km west of Yogyakarta Province, Indonesia. SUBJECTS: During the period from 1996 to 1998, up to six 24 h recalls were performed during the second trimester of pregnancy among 450 women. Nutrient intake and iron status was evaluated in relation to date of data collection relative to the economic crisis that emerged in August 1997. A computer program (Inafood) was developed to calculate nutrient intake. RESULTS: : Forty percent of the pregnant women were at risk of inadequate intake of energy and protein, and 70% were at risk of inadequate intake of vitamin A, calcium and iron even before the crisis. Our results also demonstrate an effect of short-term economic crisis on nutrient intake and iron status. When the crisis emerged, urban poor experienced a decrease in intake of most nutrients. During the crisis, rich women experienced a significant decrease in fat (P<0.05). Negative changes in fat density during crisis were experienced by the rich and the rural, poor, and access to rice field subgroups (P<0.01). A significant increase in carbohydrate densities was seen for the rich and rural, poor, and access to rice fields groups (P<0.05). Urban poor experienced decreased serum ferritin concentration (P<0.05), whereas rich women experienced a significant increase (P<0.05). CONCLUSIONS: Urban poor and rural poor landless women were most directly affected by the emerging economic crisis. PMID- 12771968 TI - Dietary calcium intake in premenopausal Bangladeshi women: do socio-economic or physiological factors play a role? AB - OBJECTIVE: Evaluation of data on dietary calcium intake in premenopausal women of two socio-economic groups in Bangladesh. DESIGN: A cross sectional study. Three days dietary records were used to estimate habitual calcium intake. SETTING: Two regions of Bangladesh. The Dhaka city area and the Betagair Union in the sub district Nandail, Mymensingh. SUBJECTS: A total of 191 subjects of two groups (low socio-economic group=group L, n=101 and high socio-economic group=group H, n=90) of Bangladeshi women aged 16-40 y. About 87% of the subjects were housewives and the rest 13% were distributed in other different professions. Each group consisted of three sub-groups (non-pregnant non-lactating=1, pregnant=2 and lactating=3). RESULTS: : The influence of socio-economic status on dietary intake of calcium (P<0.001) was observed in this study. The dietary intake of calcium was influenced by physiological status (PS) in high income group only (P<0.005). The mean dietary calcium intake was significantly higher (P&<0.005) in all sub groups of this group compared with the corresponding sub-groups in low income group. Although in group H, 47% of subjects failed to meet even the lowest level (400-500 mg/day) of WHO recommended dietary allowances (RDA) of calcium for adult women. No subject in group L was found to meet the RDA level. Moreover, 63% of the women in group L had calcium intake lower than 200 mg/day. These figure could be more critical in both groups if we consider the recent USA-RDAs of calcium for adult women (1000 mg/day). The observed sources of dietary calcium were different in the two groups. CONCLUSIONS: The results of the study suggested that low calcium intake could reduce the bone accretion rates and increase the risk of osteoporosis in the subjects of the present study. Calcium rich food may be recommended for women in both groups. PMID- 12771970 TI - Low body weight and its association with bone health and pubertal maturation in Chinese girls. AB - OBJECTIVE: To investigate the prevalence of low body weight in Beijing pubertal girls and to establish the cut-off for body mass index (BMI) for underweight for Chinese pubertal girls. DESIGN: Cross-sectional study. SETTING: Three socioeconomic areas (rural, suburban and urban) in Beijing, China. SUBJECTS: Random sample of 1214 adolescent girls aged 12-14 y from 13 middle schools. RESULTS: Using a modified Chinese reference, the rate of low body weight (BMI<18) was 32.2% (95% CI 29.6-34.8%). Compared with desirable weight girls (BMI=18-21), girls with low body weight had a lower bone age, delayed breast and pubic hair development, a lower rate of menarche, lower distal one-third radius and ulna bone mineral content (BMC), bone mineral density and bone width. Logistic regression showed that BMI was one of the predictors of one-third ulna BMC after adjustment for confounding variables. When comparing BMI<18 vs BMI=18-21, the risk of BMC being less than the median increased by 82% (odds ratio 1.82, 95% CI 1.06-3.13). Thinness and stunting rates assessed by WHO recommended cut-offs are also reported. CONCLUSIONS: High prevalence of low body weight (BMI<18) was found to be a major health problem among Beijing pubertal girls. BMI<18 is confirmed as the cut-off for delayed general growth and development for Chinese girls and for screening girls at risk of lower bone mineral status. PMID- 12771969 TI - Safety of long-term consumption of plant sterol esters-enriched spread. AB - OBJECTIVE: To evaluate both efficacy and safety in humans of long-term consumption of spreads containing plant sterol esters. DESIGN: Randomized double blind placebo-controlled parallel trial. SUBJECTS: : Hundred and eighty-five healthy volunteers (35-64 y). INTERVENTION: Volunteers daily consumed 20 g spread enriched with 1.6 g plant sterols as fatty acid esters or a control spread for 1 y. They continued their habitual diet and lifestyle. Outcome measures included efficacy markers such as total and LDL-cholesterol, a large range of safety parameters, and reporting of adverse events. RESULTS: Consumption of the plant sterol ester-enriched spread consistently lowered total and LDL cholesterol during the 1 y period on average by 4 and 6%, respectively (0.01 < P < 0.05). Plant sterols intake did on average not result in a lower carotenoid concentration (when expressed per LDL-cholesterol) after 52 weeks (P>0.05). However, carotenoid concentrations changed over time. Plant sterols intake reduced lipid adjusted alpha- and beta-carotene-concentrations by only 15-25% after 1 y, relative to control. Lipid-adjusted fat-soluble vitamin concentrations remained unchanged. Plant sterol concentrations in serum were increased from 2.76 to 5.31 ( micro mol/mmol total cholesterol) for campesterol (P<0.0001) and from 1.86 to 2.47 ( micro mol/mmol total cholesterol) for beta-sitosterol (P<0.0001). The increase in total plant sterol concentration in red blood cells (5.29-9.62 micro g/g) did not affect red blood cell deformability. Hormone levels in males (free and total testosterone) and females (luteinizing hormone, follicle stimulating hormone, beta-estradiol and progesterone) as well as all clinical chemical and hematological parameters measured were unaffected. Adverse events reported were not different between subjects consuming control spread and subjects consuming plant sterol esters-enriched spread. CONCLUSION: Consumption of a plant sterol esters-enriched spread is an effective way to consistently lower blood cholesterol concentrations and is safe to use over a long period of time. PMID- 12771971 TI - Tolerance of symptomatic lactose malabsorbers to lactose in milk chocolate. AB - OBJECTIVE: To study tolerance to lactose in milk chocolate among symptomatic lactose maldigesters. DESIGN: Randomized cross-over study. SUBJECTS: Twenty-seven adult lactose maldigesters with symptomatic lactose intolerance. METHODS: A 100 g chocolate sample prepared with whole milk (12 g lactose), whole-milk powder (12 g lactose), low-lactose milk powder (2 g lactose) or lactose-free milk powder was eaten after an overnight fast. Gastrointestinal symptoms (flatulence, abdominal bloating, abdominal pain, borgorygmi and nausea) were recorded in a questionnaire during the following 8 h. Bowel movements and stool consistency were also registered during the test day. RESULTS: The numbers of persons reporting different gastrointestinal symptoms or any of the symptoms did not differ significantly after eating the chocolate samples. No statistical differences were found in the estimated strength of the different symptoms or the total strength of all symptoms combined. Differences in the bowel frequency and stool consistency were also non-significant. CONCLUSIONS: Lactose malabsorbers with self-reported lactose intolerence did not differ in their response to milk chocolate samples containing different amounts of lactose. PMID- 12771972 TI - Copper and zinc intake and serum levels in patients with juvenile rheumatoid arthritis. AB - OBJECTIVE: To evaluate the copper and zinc intake and serum levels in patients with juvenile rheumatoid arthritis (JRA), considering the pauci and polyarticular types, the disease activity and duration, the number of inflamed joints and the use of corticosteroids therapy. DESIGN: Cross-sectional study with control group. SETTING: Outpatients of the pediatric rheumatology public health clinic, of the Universidade Federal de Sao Paulo/Escola Paulista de Medicina, Brazil. SUBJECTS: Forty-one patients with JRA were evaluated and 23 patients' brothers, as a control group. INTERVENTIONS: Copper and zinc intake evaluation by Food Register method. Copper and zinc serum levels by atomic absorption spectrophotometry. RESULTS: The disease activity did not determine difference in copper (P=0.624) and zinc (P=0.705) intake, being predominantly below the Recommended Dietary Allowances. The serum copper in relation to control was statistically greater (P=0.018), showing that the number of inflamed joints is statistically significantly related with its variation (P=0.001). The serum zinc was not different either in relation to control (P=0.940) or to the disease characteristics. CONCLUSIONS: The evaluation of copper intake seems to be of fundamental importance. It may influence the efficiency of the organic serum response. More research is needed to indicate, with security, adequate zinc intake. PMID- 12771973 TI - Influence of fish oil supplementation on in vivo and in vitro oxidation resistance of low-density lipoprotein in type 2 diabetes. AB - OBJECTIVE: Fish oil supplement has been proposed as a non-pharmacological strategy to correct the atherogenic lipid profile associated with type 2 diabetes mellitus. However, fish oil may have deleterious effects on lipid peroxidation and glycemic control. DESIGN: In this study, 44 type 2 diabetic patients were randomized to vitamin E standardized (53.6 mg/day) supplementation (capsules) with 4 g daily of either fish oil (n=23) or corn oil (n=21) for 8 weeks preceded by a 4 week run-in period of corn oil supplementation. LDL was isolated by density gradient ultracentrifugation and oxidized in vitro with Cu(2+). As a marker of in vivo oxidation malondialdehyde concentration in LDL (LDL-MDA) was measured. RESULTS: Fish oil reduced both mean lag time (before, 57.8; after, 48.8 min, P<0.001) and mean propagation rate (before, 0.018 DeltaOD/min; after, 0.015 DeltaOD/min, P<0.001), whereas corn oil had no influence on lag time and propagation rate. The changes in lag time and propagation rate differed significantly between fish oil and corn oil treatment. LDL-MDA changes differed borderline significantly between groups (FO, 110.4 pmol/mg protein; CO, 6.7 pmol/mg protein; P=0.057). Fish oil supplementation had no influence on glycemic control as assessed from HbA(1c) and fasting blood glucose. CONCLUSION: According to our findings, fish oil supplementation leads to increased in vivo oxidation and increased in vitro oxidation susceptibility of LDL particles. More studies are needed to clarify the clinical importance of this finding. PMID- 12771974 TI - Chitosan decreases total cholesterol in women: a randomized, double-blind, placebo-controlled trial. AB - BACKGROUND: Hypercholesterolemia is an important risk factor for cardiovascular disease. Orally administered chitosan binds lipids in the small intestine and reduces their absorption. Chitosan has been shown to decrease serum cholesterol in animal and human studies. This study investigated the effectiveness of chitosan in reducing serum cholesterol without concomitant diet therapy. METHODS: Ninety female volunteers (age 34-70 y) with confirmed mild to moderate hypercholesterolemia were enrolled into the study. They were randomly assigned to receive chitosan (1.2 g per day) or placebo in a double-blind manner. Serum lipids, body weight and adverse events were assessed at baseline and after 28 and 56 days of treatment. Subjects maintained their usual diet and documented the type and gross amount of food consumed. RESULTS: Eighty-four subjects (41 chitosan, 43 placebo) were included in the analysis. Chitosan significantly (F=3.19, P=0.04) reduced total cholesterol compared to placebo. In a subgroup of subjects with over 60 y of age, chitosan group significantly reduced total and LDL cholesterol (F=4.21, P=0.02, and F=3.46, P=0.04, respectively) compared with placebo. Adverse effects were few; no serious events were reported. CONCLUSION: Our results demonstrate that chitosan is safe and effective for lowering cholesterol. However, the effect of chitosan for decreasing cholesterol is mild. PMID- 12771976 TI - Recruiting children into cancer trials--role of the United Kingdom Children's Cancer Study Group (UKCCSG). AB - The UK Children's Cancer Study Group (UKCCSG), established in 1977, provides a highly organised structure for both service provision and research, and represents the model to which the adult cancer community is currently aspiring. Since childhood cancer is so rare, it is both essential and feasible for the vast majority of children to be referred into the network of specialist centres, and also for the maximum number of children to be recruited into national and international clinical trials. Over the last 30-40 years there have been major advances in treatment, such that now approximately 70% of children diagnosed with cancer will be cured of their disease. The conduct of clinical trials in this patient population does, however, raise a number of specific issues and these are discussed in the paper. PMID- 12771977 TI - Persistent excess mortality from lung cancer in patients with stage I non-small cell lung cancer, disease-free after 5 years. AB - Among patients with non-small-cell lung cancer (NSCLC), those with pathological stage I have the best expectation of survival; however, survival is reduced to less than 50% in the long term. At present, it is unclear when patients can be reasonably defined as cured, and if they experience a higher incidence of malignant/nonmalignant diseases and a lower expectation of survival than the general population. A total of 134 stage I NSCLC patients, who had undergone resection at the Thoracic Surgery Unit of the General Hospital of Verona (north eastern Italy) from October 1987 to December 1993, were still disease-free at 5 years. These subjects were further followed up, and morbidity and mortality rates were compared with those recorded in the general population of the same geographical area. The standardised incidence ratios (SIRs) for all malignancies and for lung cancer were higher than expected (2.39, 95% CI=1.6-3.5, P<0.001; 10.1, 95% CI=6.2-15.6, P<0.0001, respectively). The standardised mortality ratio (SMR) was also significantly increased (1.73, 95% CI=1.1-2.6, P=0.013). The excess mortality could be entirely explained by an increase in mortality from lung cancer (5.7, 95% CI=2.8-10.1, P<0.0001). This study shows that patients, resected for pathological stage I NSCLC and tumour-free after 5 years, have a higher incidence of new lung cancer compared with the general population, which in turn determines an excess in all-cause mortality in the following years. PMID- 12771978 TI - Phase II multicentre study of docetaxel plus 5-fluorouracil in patients with anthracycline-pretreated metastatic breast cancer. AB - The purpose of the study was to determine the efficacy and safety of docetaxel plus continuous infusion of 5-fluorouracil (5-FU) in patients with metastatic breast cancer previously treated with anthracyclines. A total of 41 patients with histologically proven metastatic breast cancer and performance status 0-2, who had received at least one anthracycline-containing regimen, received docetaxel 85 mg m(-2) followed by continuous infusion of 5-FU 750 mg m(-2) day(-1) for 5 days every 3 weeks for up to eight cycles. All patients received corticosteroid premedication, but there was no prophylactic colony-stimulating factor support. The most frequent metastatic sites were the liver (61%), bone (29%), and lung (29%). All 41 patients were assessable for toxicity and 30 were eligible and assessable for efficacy. The objective response rate was 70.0% (95% CI: 53.6 86.4%) for the per protocol group and 53.7% (95% CI: 38.4-68.9%) for the intent to-treat (ITT) population. For the ITT population, median duration of response was 8.4 months (95% CI: 6.7-12.2 months), median time to progression was 6.7 months (95% CI 5.5-8.6 months), and median survival was 17 months (95% CI: 12.3 not recorded months). Grade 3/4 neutropenia occurred in 54% of patients, with febrile neutropenia in 24% of patients and 5% of cycles, but infections were rare. Stomatitis was frequent, grade 3 in 24% of patients and grade 4 in one patient (2%), but manageable. Diarrhoea was rare, grade 3 in 7% of patients and 1% of cycles. Other grade 3/4 nonhaematological toxicities were infrequent. In conclusion, this docetaxel/5-FU regimen is highly active and well tolerated in patients with anthracycline-pretreated metastatic breast cancer. The efficacy is particularly promising, as one-third of patients were either second-line and/or anthracycline-resistant/refractory. PMID- 12771979 TI - Genetic counselling and the intention to undergo prophylactic mastectomy: effects of a breast cancer risk assessment. AB - Scientific reports suggest that women at risk for familial breast cancer may benefit from prophylactic mastectomy. However, few data are available about how women decide upon this clinical option, and in particular, what role an objective risk assessment plays in this. The purpose of the present study is to assess whether this objective risk information provided in genetic counselling affects the intention for prophylactic mastectomy. Additionally, the (mediating) effects of breast cancer worry and perceived risk are investigated. A total of 241 women completed a questionnaire before and after receiving information about their familial lifetime breast cancer risk in a genetic counselling session. Path analysis showed that the objective risk information had a corrective effect on perceived risk (beta=0.38; P=0.0001), whereas the amount of breast cancer worry was not influenced by the counselling session. The objective risk information did not directly affect the intention for prophylactic mastectomy. The intention was influenced by perceived risk after counselling (beta=0.23; P=0.002), and by the precounselling levels of perceived risk (beta=0.27; P=0.00025) and breast cancer worry (beta=0.32; P=0.0001), that is, higher levels of perceived risk and breast cancer worry imply a stronger intention for prophylactic mastectomy. A personal history of breast cancer did not directly influence the intention for prophylactic mastectomy, but affected women who had undergone a mastectomy as surgical treatment were more positively inclined to have a prophylactic mastectomy than women who had had breast-conserving therapy. The impact of objective risk information on the intention for prophylactic mastectomy is limited and is mediated by perceived risk. Important determinants of the intention for prophylactic mastectomy were precounselling levels of breast cancer worry and perceived risk, suggesting that genetic counselling is only one event in the entire process of decision making. Therefore, interventions aimed at improving decision making on prophylactic mastectomy should explicitly address precounselling factors, such as personal beliefs and the psychological impact of the family medical history. PMID- 12771980 TI - Are diet-prostate cancer associations mediated by the IGF axis? A cross-sectional analysis of diet, IGF-I and IGFBP-3 in healthy middle-aged men. AB - We examined the association of diet with insulin-like growth factors (IGF) in 344 disease-free men. Raised levels of IGF-I and/or its molar ratio with IGFBP-3 were associated with higher intakes of milk, dairy products, calcium, carbohydrate and polyunsaturated fat; lower levels with high vegetable consumption, particularly tomatoes. These patterns support the possibility that IGFs may mediate some diet cancer associations. PMID- 12771982 TI - Up-to-date survival curves of children with cancer by period analysis. AB - Survival rates of children with cancer have strongly improved during the past decades, but much of this improvement has been disclosed with substantial delay by traditional methods of survival analysis, which reflect survival experience of patients diagnosed many years ago. In this paper, the use of a new method of survival analysis, denoted period analysis, for providing more up-to-date estimates of 10-year survival curves of children with cancer is empirically evaluated using data of the Surveillance, Epidemiology, and End Results Program of the United States National Cancer Institute. It is shown that period analysis provides much more up-to-date estimates of survival curves than traditional cohort-based survival analysis indeed, at least as long as there is ongoing improvement in survival rates over time, as it seems to be the case for many forms of childhood cancer. The most recent 10-year period survival estimates indicate that survival rates of children with cancer achieved by the end of the 20th century are substantially higher than previously available survival statistics have suggested. Application of period analysis may be particularly useful in the field of childhood cancer as it may help to prevent patients, their families and clinicians from being burdened by outdated, often too pessimistic survival expectations. PMID- 12771981 TI - Risk of cancer in a large cohort of nonaspirin NSAID users: a population-based study. AB - There is increasing evidence of an inverse association between use of nonsteroidal anti-inflammatory drugs (NSAIDs) and risk of colorectal cancer. However, data regarding other cancer sites are limited. Using data from the population-based North Jutland Prescription Database and the Danish Cancer Registry, we compared cancer incidence among 172 057 individuals prescribed nonaspirin NSAIDs with expected incidence (based on county-specific cancer rates) during a 9-year study period. A total of 6081 incident cancer cases were diagnosed among NSAID users vs 5722 expected (standardised incidence ratio (SIR) 1.1, 95% confidence interval (CI)1.0-1.1). The SIRs for colon and rectal cancer among persons who obtained 10 or more prescriptions were 0.7 (95% CI 0.6-0.9) and 0.6 (95% CI 0.4-0.9), respectively. Similarly, reduced risk estimates were found for stomach (SIR 0.7, 95% CI 0.4-1.1) and ovarian cancer (SIR 0.7, 95% CI 0.4 1.0). Standardised incidence ratios for other cancers among those with 10 or more prescriptions tended to be close to 1.0, except for lung, kidney, and prostate cancers with SIRs of 1.3 (95% CI 1.1-1.6), 1.4 (95% CI 0.9-2.1), and 1.6 (95% CI 1.3-2.0), respectively. We found protective associations of NSAIDs against colon, rectal, stomach, and ovarian cancer. Reasons for the increased risk for some cancer sites are not clear. PMID- 12771983 TI - Cancer survival in parents who lost a child: a nationwide study in Denmark. AB - Psychological stress has been suggested to shorten cancer survival, but few studies have examined the effect of parental bereavement, and the results have been inconsistent. We identified all 21 062 parents who lost a child in Denmark from 1980 to 1996 and among them, 1630 parents with subsequent incident cancer formed the exposed cohort. We recruited 6237 incident cancer patients from a group of 293 745 randomly selected unexposed parents matched on family structure at the same time as the bereaved parents. All incident cancers in the two cohorts were followed to the end of 1997, or until they died. Cox proportional-hazards regression models were used to evaluate the hazard ratio (HR) of dying in exposed parents with cancer. The overall HR of dying from an incident cancer in exposed parents was 1.23 (95% confidence interval 1.03-1.47) compared to parents with cancer who did not lose a child. The HRs were nearly identical to those in the unexposed parents for site-specific cancers like lung cancer, breast cancer, and other groups of cancers like cancers in all digestive organs, smoking-related cancers, alcohol-related cancers, hormone-related cancers, virus/immune-related cancers, and lymphatic/haematopoietic cancers. Death of a child is not a strong prognostic factor for cancer survival among parents diagnosed with cancer after the bereavement. However, a small impairment in overall cancer survival cannot be ruled out. PMID- 12771985 TI - Specialisation and breast cancer survival in the screening era. AB - It is recommended that specialist surgeons treat all breast cancer, although the limited evidence to support this is based on treatment patterns prior to the introduction of screening. Whether a specialist survival advantage exists in the post-screening era is uncertain, as referral and treatment patterns may have changed, in addition to the effect of screening on the natural history of breast cancer. Our aim was to determine the impact of screening on the caseload and case mix of specialist surgeons, to determine if the survival advantage associated with specialist care is maintained with longer follow-up and persists after the introduction of screening. Using the West of Scotland Cancer Registry, all 7197 women treated for breast cancer in a 15-year time period (1980-1994) in a geographically defined cohort were followed up for an average of 9 years, and pathological stage and socioeconomic status were linked with mortality data. We show that the caseload of specialists has increased substantially (from 11 to 59% of the total workload) and that smaller cancers have been selectively referred. However, even after allowing for pathological stage, socioeconomic status and method of detection, specialist treatment was associated with a significantly lower risk of dying (prescreening: relative risk of dying=0.83, 95% CI=0.75-0.92; post-screening: relative risk of dying=0.89, 95% CI=0.78-1.00). We conclude that this survival benefit is most consistent with effective surgical management rather than selective referral, the influx of screen-detected cancers or adjuvant therapies. PMID- 12771984 TI - Lifestyles, environmental and phenotypic factors associated with lip cancer: a case-control study in southern Spain. AB - The aim of this study was to identify factors related to lip cancer (LC) considering individual characteristics and sociodemographic factors. A case control study was carried out in the province of Granada (Andalusia, southern Spain). The cases were 105 males with squamous-cell carcinoma of the lip, diagnosed between 1987 and 1989 (aged 20-70 years) and identified by means of a population-based Cancer Registry. As controls, a randomised populational sample of 239 males, stratified by age, was used. Multiple logistic regression analysis showed that risk factors are lifetime cumulative tobacco consumption and alcohol consumption. An interaction was found between alcohol consumption and the smoking habit (leaving the cigarette on the lip): OR=23.6; 95% CI: 3.9-142.0. Other risk factors identified are clear eyes (OR=3.5; CI: 95% 1.5-8.0), sun exposure early in life and cumulative sun exposure during outdoor work (OR=11.9; 95%: CI: 1.3 108.9), and skin reaction to sun exposure (Fitzpatrick levels). Another interaction was found between skin reaction and a previous history of common sporadic warts (OR=4.4; 95% CI: 1.01-19.1). We conclude that LC is related to phenotype, skin reaction to sun exposure, cumulative and early sunlight exposure, and tobacco and alcohol consumption, as well as a low educational level. Leaving the cigarette on the lip is predictive of LC risk irrespective of cumulative tobacco consumption. PMID- 12771986 TI - Human papillomavirus infection and use of oral contraceptives. AB - Human papillomavirus (HPV) infection is thought to be a necessary but not sufficient cause of most cases of cervical cancer. Since oral contraceptive use for long durations is associated with an increased risk of cervical cancer, it is important to know whether HPV infection is more common in oral contraceptive users. We present a systematic review of 19 epidemiological studies of the risk of genital HPV infection and oral contraceptive use. There was no evidence for a strong positive or negative association between HPV positivity and ever use or long duration use of oral contraceptives. The limited data available, the presence of heterogeneity between studies and the possibility of bias and confounding mean, however, that these results must be interpreted cautiously. Further studies are needed to confirm these findings and to investigate possible relations between oral contraceptive use and the persistence and detectability of cervical HPV infection. PMID- 12771988 TI - Neural-cadherin expression associated with angiogenesis in non-small-cell lung cancer patients. AB - An immunohistochemical analysis for E(epithelial)-cadherin and N(neural)-cadherin expression in relation to tumour angiogenesis was performed in 150 patients with nonsmall cell lung cancer (NSCLC). In all, 71 carcinomas (47.3%) were E-cadherin negative. Epithelial-cadherin-negative tumours had lymph node metastases significantly more frequently than E-cadherin-positive tumours (P=0.0100). On the other hand, 46 carcinomas (30.7%) were N-cadherin-positive. Regarding tumour vascularity, there was no significant correlation between E-cadherin expression and tumour vascular. In contrast, the frequency of hypervascular tumours was significantly higher for N-cadherin-positive carcinomas than for N-cadherin negative carcinomas (P=0.0373). Regarding prognosis, the 5-year survival rate of patients with E-cadherin-negative NSCLCs was significantly lower than that of patients with E-cadherin-positive NSCLCs (P=0.0146). In contrast, of the patients with large cell carcinomas, the 5-year survival rate of patients with N-cadherin positive tumours was significantly lower than that of patients with N-cadherin negative tumours (P=0.0013). A multivariate analysis demonstrated that E-cadherin status (P=0.0339) and tumour vascularity (P=0.0295) were significant indicators for survival. In conclusion, E-cadherin expression and tumour vascularity are significant prognostic factors of NSCLC patients. Furthermore, N-cadherin expression is associated with tumour angiogenesis, and its expression is one of prognostic factors of patients with large cell carcinomas. Thus, N-cadherin also might play a specific role in undifferentiated large cell carcinomas. PMID- 12771987 TI - Prognostic value of serum levels of interleukin 6 and of serum and plasma levels of vascular endothelial growth factor in hormone-refractory metastatic breast cancer patients. AB - Prediction of survival for patients with metastatic breast cancer is often inaccurate and may be helped by new biological parameters. Tumour growth being angiogenesis-dependent, it has been hypothesised that the assessment of angiogenic factor production might reflect the clinical behaviour of cancer progression. This study was designed to investigate the clinical significance of vascular endothelial growth factor (VEGF) and interleukin 6 (IL-6) in hormone refractory metastatic breast cancer. Serum and plasma concentrations of VEGF and serum concentration of IL-6 were measured in 87 patients with a fully documented history of metastatic breast cancer using an enzyme-linked immunoassay. All patients had detectable levels of VEGF, whereas 39% patients had detectable serum levels of IL-6. There was a positive correlation between IL-6 levels and the theoretical VEGF load of platelets (P<0.001). The presence of high levels of serum IL-6, but not VEGF, was significantly correlated to a shorter survival. In a multivariate analysis along with clinical prognostic parameters, serum IL-6 was identified as an independent adverse prognostic variable for overall survival (P&<0.001). These results indicate that serum IL-6 levels correlate to poor survival in patients with hormone-refractory metastatic breast cancer. Vascular endothelial growth factor serum and plasma levels are not useful indicators of prognosis for these patients. PMID- 12771989 TI - Reduced expression of Axin correlates with tumour progression of oesophageal squamous cell carcinoma. AB - Axin is a negative regulator of the Wnt signalling pathway, and genetic alterations of AXIN1 have been suggested to be an important factor of carcinogenesis in some tumours. The objective of this study was to clarify the clinicopathologic and prognostic significance of Axin in oesophageal squamous cell carcinoma (SCC). Immunohistochemical staining for Axin was performed on surgical specimens obtained from 81 patients with oesophageal SCC. Western and Northern blottings were performed on proteins and RNA from oesophageal SCC cell lines. Then polymerase chain reaction-single-strand conformational analysis (PCR SSCP) was performed on DNA from oesophageal SCC patients and cell lines. Axin expression was found to be correlated inversely with depth of invasion, lymph node metastasis, and lymphatic invasion. Although univariate analysis showed Axin to be a negative predictor, multivariate analysis showed that it was not an independent prognostic marker. In all but one of the seven cell lines examined, the levels of protein expression were equivalent to RNA expression. PCR-SSCP showed that five patients and three cell lines had polymorphisms in exon 4 or 5 of the AXIN1 gene, but none of the 81 patients with oesophageal SCC had mutations. Our findings suggest that reduced expression of Axin is correlated with tumour progression of oesophageal SCC. However, additional studies will be necessary to elucidate the mechanism responsible for loss of Axin expression in tumour cells. PMID- 12771990 TI - Cytology vs molecular analysis for the detection of head and neck squamous cell carcinoma in oesopharyngeal brush samples: a prospective study in 56 patients. AB - Oesopharyngeal brush (OPB) sampling with cytological analysis can yield exfoliated cells from asymptomatic tumours of the upper aero-digestive tract and the oesophagus. In this study, we compared cytological evaluation and molecular analysis for the detection of exfoliated cancer cells sampled with an OPB. A total of 56 patients with a known unique head and neck squamous cell carcinoma (HNSCC) and five healthy controls were enrolled prospectively. Exfoliated cells from these 61 patients were collected with an OPB before initial endoscopy. p53 mutations and UT 5085 microsatellite instability (MI) were analysed in the HNSCC tumour, lymphocytes and the corresponding OPB DNA samples. p53 mutations and UT5085 MI were detected in 31 out of 56 and 14 out of 56 HNSCC, respectively, but not in any of the five controls. Direct sequencing of p53 was able to detect mutations in OPB DNA in only two out of 29 patients harbouring a p53-mutated primary tumour. Microsatellite instability was detected in OPB DNA of 11 out of 13 informative (bandshift detected in tumour) patients, whereas cytological analysis detected abnormal cells in only six of the same 13 patients (P=0.03). In informative patients, all positive OPB samples at cytological analysis were also positive at molecular analysis of UT5085, and both analyses confirmed the two negative samples. Molecular analysis of OPB from eight uninformative patients and from five healthy controls were all negative. OPB sampling with MI-based molecular analysis could be efficient for early detection of recurrent HNSCC. This result prompts us to use other microsatellite markers in order to maximise the percentage of informative patients. PMID- 12771991 TI - NESP55, a novel chromogranin-like peptide, is expressed in endocrine tumours of the pancreas and adrenal medulla but not in ileal carcinoids. AB - Neuroendocrine secretory protein 55, NESP55, is an acidic protein belonging to the chromogranin family. The distribution of NESP55 in human tumours is not known. The aim of the present study was to study the expression of NESP55 in human gastrointestinal, pancreatic and adrenal tumours. A total of 118 human endocrine and nonendocrine tumours were examined by immunocytochemistry, and compared to the expression of chromogranin A (CgA) in the same tumours. Pancreatic endocrine tumours (14 out of 25), pheochromocytomas (19 out of 19), and neuroblastomas (seven out of 14) expressed NESP55, with the same strong labelling pattern in both benign and malignant tumours. Expression of NESP55 in pancreatic endocrine tumours and pheochromocytomas was confirmed by Western and Northern blot analysis. Immunocytochemical analysis demonstrated no labelling in ileal carcinoids (zero out of 15), and adrenocortical adenomas (zero out of 15). The majority of gastrointestinal and pancreatic carcinomas were negative for NESP55, with focal staining observed in two out of 30 tumours. In contrast, CgA was present in all neuroendocrine tumours examined (25 out of 25 pancreatic endocrine tumours, 19 out of 19 pheochromocytomas, 14 out of 14 neuroblastomas and 15 out of 15 ileal carcinoids). Thus, the expression of NESP55 in endocrine tumours of the gastrointestinal tract, pancreas and adrenals differs from that of CgA. Neuroendocrine secretory protein 55 is found in a subset of neuroendocrine tumours showing differentiation towards adrenal chromaffin cells and pancreatic islets cells. PMID- 12771992 TI - Characterisation of integrin-linked kinase signalling in sporadic human colon cancer. AB - The putative oncogene, integrin-linked kinase (ILK) is a protein serine/threonine kinase that has been reported to regulate a number of biological properties including anchorage-independent cell cycle progression, tumour cell invasion and apoptosis. Overexpression of ILK has been documented in a wide variety of human malignancies including Ewing's sarcoma (ES), primitive neural ectodermal tumours (PNETs) and prostate tumours (PT). We recently reported that ILK signalling was also dysregulated in patients with the genetic condition familial adenomatous polyposis (FAP), a precursor to colon cancer. In this study, we extended our previous work by investigating the ILK-signalling pathway in sporadic human colon cancer and representative lymph node metastases. The data indicate that the ILK protein is significantly hyperexpressed in malignant acini in relation to normal crypts. Moreover, overexpression of ILK not only coincided with increased MBP phosphotransferase activity but as well with effects on downstream targets like GSK3beta. Based upon the presented data, we propose that ILK signalling is dysregulated early during the development of human colon cancer, and that selective inhibition of this molecule alone or in combination with the standard therapeutic modality might be a more effective means of treating colon cancer. PMID- 12771993 TI - In vivo evolution of tumour cells after the generation of double-strand DNA breaks. AB - In vitro, the ratio of single- to double-strand DNA breaks (DSB) and their absolute values determine the cell death pathway. The consequences of the generation of various numbers of DSB generated in vivo in tumour cells have been analysed in two different experimental tumour models. Synchronisation of DSB generation and control of their number have been achieved using different doses of bleomycin (BLM) and tumour cell permeabilisation by means of locally delivered electric pulses. According to BLM dose, different cell death pathways are observed. At a low therapeutic dose, a mitotic cell death pathway is detected. It is characterised by the appearance of 'atypical mitosis', TUNEL and caspase-3 positive, 24 h after the treatment, and later by the presence of typical apoptotic figures, mainly TUNEL positive but caspase-3 negative. Caspase-3 is thus an early marker of apoptosis. Mitotic cell death is also followed by lymphocytic infiltration reaction. At high doses of BLM, pseudoapoptosis is detected within a few minutes after the treatment. These cell death pathways are discussed as a function of the number of DSB generated, by comparison with previous results obtained in vitro using BLM or ionising radiation. PMID- 12771994 TI - Role of cytokines in photodynamic therapy-induced local and systemic inflammation. AB - Photodynamic therapy (PDT) of tumour results in the rapid induction of an inflammatory response that is considered important for the activation of antitumour immunity, but may be detrimental if excessive. The response is characterised by the infiltration of leucocytes, predominantly neutrophils, into the treated tumour. Several preclinical studies have suggested that suppression of long-term tumour growth following PDT using Photofrin((R)) is dependent upon the presence of neutrophils. The inflammatory pathways leading to the PDT-induced neutrophil migration into the treated tumour are unknown. In the following study, we examined, in mice, the ability of PDT using the second-generation photosensitiser 2-[1-hexyloxyethyl]-2-devinyl pyropheophorbide-a (HPPH) to induce proinflammatory cytokines and chemokines, as well as adhesion molecules, known to be involved in neutrophil migration. We also examined the role that these mediators play in PDT-induced neutrophil migration. Our studies show that HPPH PDT induced neutrophil migration into the treated tumour, which was associated with a transient, local increase in the expression of the chemokines macrophage inflammatory protein (MIP)-2 and KC. A similar increase was detected in functional expression of adhesion molecules, that is, E-selectin and intracellular adhesion molecule (ICAM)-1, and both local and systemic expression of interleukin (IL)-6 was detected. The kinetics of neutrophil immigration mirrored those observed for the enhanced production of chemokines, IL-6 and adhesion molecules. Subsequent studies showed that PDT-induced neutrophil recruitment is dependent upon the presence of MIP-2 and E-selectin, but not on IL 6 or KC. These results demonstrate a PDT-induced inflammatory response similar to, but less severe than obtained with Photofrin((R)) PDT. They also lay the mechanistic groundwork for further ongoing studies that attempt to optimise PDT through the modulation of the critical inflammatory mediators. PMID- 12771995 TI - Photodynamic diagnosis of ovarian cancer using hexaminolaevulinate: a preclinical study. AB - The unfailing detection of micrometastases during surgery of patients suffering from ovarian cancer is mandatory for the optimal management of this disease. Thus, the present study aimed at determining the feasibility of detecting micrometastases in an ovarian cancer model using the intraperitoneal administration of the photosensitiser precursor hexaminolaevulinate (HAL). For this purpose, HAL was applied intraperitoneally at different concentrations (4-12 mM) to immunocompetent Fischer 344 rats bearing a syngeneic epithelial ovarian carcinoma. The tumours were visualised laparoscopically using both white and blue light (D-light, Karl Storz, Tuttlingen, Germany), and the number of peritoneal micrometastases detected through HAL-induced photodiagnosis (PD) was compared to standard white light visualisation. Fluorescence spectra were recorded with an optical fibre-based spectrofluorometer and the fluorescence intensities were compared to the protoporphyrin IX (PpIX) fluorescence induced by 5-aminolevulinic acid under similar conditions. The number of metastases detected by the PD blue light mode was higher than when using standard white light abdominal inspection for all applied concentrations. Twice as many cancer lesions were detected by fluorescence than by white light inspection. The hexyl-ester derivative produced higher PpIX fluorescence than its parent substance aminolevulinic acid at the same concentration and application time. Fluorescence contrast between healthy and cancerous tissue was excellent for both compounds. To overcome poor diagnostic efficiency and to detect peritoneal ovarian carcinoma foci in the large surface area of the human peritoneal cavity, HAL fluorescence-based visualisation techniques may acquire importance in future and lead to a more correct staging of early ovarian cancer. PMID- 12771996 TI - Recombinant mistletoe lectin induces p53-independent apoptosis in tumour cells and cooperates with ionising radiation. AB - Mistletoe extracts are used as alternative cancer treatment in addition to standard chemotherapy and radiation treatment and have an immunostimulatory and pain-relieving effect. A direct antitumour effect of mistletoe extracts against tumour cells of lymphoid origin has been linked to the D-galactoside-specific mistletoe lectin I. In this study, we investigated the cellular effect of bacterially expressed, recombinant mistletoe lectin alone or in combination with ionising radiation in a genetically defined p53-wild-type and p53-deficient E1A/ras-transformed murine tumour cells system. Downregulation of the proliferative activity and cell killing by recombinant mistletoe lectin occurred in a clear dose response (0.1-1 ng ml(-1)). Induction of apoptosis was p53 independent, but apoptosis-associated factor-1-dependent. Cellular treatment with lectin in combination with ionising radiation resulted in both p53-wild-type and p53-deficient tumour cells in an at least additive, antiproliferative effect and enhanced activation of caspase-3. Combined treatment with ionising radiation and lectin revealed a similar cytotoxic effect in human, p53-mutated adenocarcinoma cells. Thus, recombinant mistletoe lectin alone and in combination with ionising radiation bypasses often prevalent apoptotic deficiencies in treatment-resistant tumour cells. PMID- 12771998 TI - Apoptosis induction in renal cell carcinoma by TRAIL and gamma-radiation is impaired by deficient caspase-9 cleavage. AB - TNF-related apoptosis-inducing ligand (TRAIL APO-2L) is a member of the TNF family and induces apoptosis in cancer cells without affecting most non neoplastic cells. The present investigation is focused on apoptosis induction by combined exposure to TRAIL and ionising radiation (IR) in human renal cell carcinoma (RCC) cell lines. Here, we demonstrate that all RCC cell lines coexpress TRAIL and the death-inducing receptors, TRAIL-R1 and TRAIL-R2. Exposure to TRAIL alone induced marked apoptosis in three out of eight RCC cell lines. Combined exposure to TRAIL and IR resulted in a sensitisation to TRAIL-induced apoptosis in one RCC cell line only. Enhanced apoptosis induction by TRAIL in combination with IR was paralleled by an increase in PARP cleavage and activation of executioner caspase-3, whereas caspases-6 and -7 were not involved. Moreover, exposure to TRAIL and/or IR resulted in a marked activation of initiator caspase 8, possibly augmented by the observed reduction of inhibitory c-FLIP expression. In contrast to other tumour types, activation of initiator caspase-9 was not detectable in our RCC model system after exposure to TRAIL and/or IR. This lack of caspase-9 activation might be related to an impaired 'crosstalk' with the caspase-8 pathway as suggested by the missing Bid cleavage and to the appearance of an XIAP cleavage product known to inhibit caspase-9 activation. Deficient activation of caspase-9, therefore, might contribute to the clinically known resistance of human RCC against IR and also argues against an effective combination therapy with TRAIL and IR in this tumour type. PMID- 12771999 TI - Increased gastrin-releasing peptide (GRP) receptor expression in tumour cells confers sensitivity to [Arg6,D-Trp7,9,NmePhe8]-substance P (6-11)-induced growth inhibition. AB - [Arg(6),D-Trp(7,9),N(me)Phe(8)]-substance P (6-11) (SP-G) is a novel anticancer agent that has recently completed phase I clinical trials. SP-G inhibits mitogenic neuropeptide signal transduction and small cell lung cancer (SCLC) cell growth in vitro and in vivo. Using the SCLC cell line series GLC14, 16 and 19, derived from a single patient during the clinical course of their disease and the development of chemoresistance, it is shown that there was an increase in responsiveness to neuropeptides. This was paralleled by an increased sensitivity to SP-G. In a selected panel of tumour cell lines (SCLC, non-SCLC, ovarian, colorectal and pancreatic), the expression of the mitogenic neuropeptide receptors for vasopressin, gastrin-releasing peptide (GRP), bradykinin and gastrin was examined, and their sensitivity to SP-G tested in vitro and in vivo. The tumour cell lines displayed a range of sensitivity to SP-G (IC(50) values from 10.5 to 119 microM). The expression of the GRP receptor measured by reverse transcriptase-polymerase chain reaction, correlated significantly with growth inhibition by SP-G. Moreover, introduction of the GRP receptor into rat-1A fibroblasts markedly increased their sensitivity to SP-G. The measurement of receptor expression from biopsy samples by polymerase chain reaction could provide a suitable diagnostic test to predict efficacy to SP-G clinically. This strategy would be of potential benefit in neuropeptide receptor-expressing tumours in addition to SCLC, and in tumours that are relatively resistant to conventional chemotherapy. PMID- 12771997 TI - Drug resistance associated with loss of p53 involves extensive alterations in microtubule composition and dynamics. AB - In the present study, we compared the dynamics and composition of microtubules in cell lines derived from the human breast adenocarcinoma MCF-7 containing either the wild-type p53 (wt-p53; MN1) or a dominant-negative variant of p53 gene (mut p53; MDD2). Mut-p53 cells were significantly resistant to the cytotoxicity of the microtubule-targeted drugs (vinca alkaloids and taxanes), as compared with wt-p53 cells. Studies by high-resolution time-lapse fluorescence microscopy in living cells indicated that the dynamics of microtubules of mut-p53 cells were altered in complex ways and were significantly increased as compared with microtubules in wt-p53 cells. The percentage of time microtubules spent in growing and shortening phases increased significantly, their catastrophe frequency increased, and their overall dynamicity increased by 33%. In contrast, their shortening rate and the mean length shortened decreased. Cells containing mut-p53 displayed increased polymerisation of tubulin, increased protein levels of the class IV beta-tubulin isotype, STOP and survivin, and reduced protein levels of class II beta-tubulin isotype, MAP4 and FHIT. We conclude that p53 protein may contribute to the regulation of microtubule composition and function, and that alterations in p53 function may generate complex microtubule-associated mechanisms of resistance to tubulin-binding agents. PMID- 12772000 TI - An alternative, non-intrauterine hypothesis, based on maternal mitochondrial oocyte inheritance, to explain inconsistent findings of birth weight on (breast) cancer risk. PMID- 12772004 TI - Validation of the GSFQ, a self-administered symptom frequency questionnaire for patients with gastroesophageal reflux disease. AB - BACKGROUND: Although the diagnosis of gastroesophageal reflux disease (GERD) is based primarily on symptoms experienced by a patient, relatively little attention has been paid to the development and validation of self-administered questionnaires specific to GERD symptoms. The present article presents the validation of the short, self-administered GERD Symptom Frequency Questionnaire (GSFQ). METHODS: Patients with GERD participating in a randomized clinical trial comparing pantoprazole and nizatidine were asked to complete the GSFQ together with validated instruments for measurement of health-related quality of life (Medical Outcome Study Short Form 12) and gastrointestinal symptoms (Gastrointestinal Symptom Rating Scale). Completion of the GSFQ, Medical Outcome Study Short Form 12 and Gastrointestinal Symptom Rating Scale took place upon entry into the trial (baseline) and during the trial (days 7 and 28). Endoscopy was performed at baseline and after 28 days. Cronbach alpha was used to assess the internal consistency of the questionnaire. The test-retest reliability of the GSFQ was examined by the intraclass correlation coefficient among the 36 patients with stable GERD symptoms between day 7 and day 28. Construct validity was assessed by comparing the GSFQ with previously validated instruments. Known group validity was determined by comparing GSFQ scores across groups of patients known to differ clinically. Responsiveness to change was assessed by the Guyatt's statistic. RESULTS: Two hundred twenty-one patients formed the study baseline group. The analysis demonstrated that the GSFQ questionnaire had excellent psychometric properties shown by the high internal consistency (Cronbach alpha 0.84); that the test-retest reliability was satisfactory (intraclass correlation coefficient 0.64); that there was good evidence that the GSFQ indeed measured what it was intended to measure (validity); and that the GSFQ was highly responsive to change (Guyatt's statistic 1.48). DISCUSSION: The GSFQ is a short, self-administered, easy to use, GERD-specific questionnaire which should be considered as a useful assessment tool in the evaluation of patients with GERD and in the assessment of treatment outcomes. PMID- 12772005 TI - A survey of Canadian gastroenterologists about the management of Barrett's esophagus. AB - The aims of the present study were to determine practice patterns of Canadian gastroenterologists for screening patients with Barrett's esophagus and to compare current practice patterns with published guidelines. A secondary goal was to evaluate whether gastroenterologists recommend a "once in a lifetime" endoscopy for patients with chronic gastroesophageal reflux disease. A structured questionnaire regarding screening for Barrett's esophagus was sent to members of the Canadian Association of Gastroenterology. The overall response rate was 51% (203 of 396). Of the 203 respondents, 165 (81%) performed endoscopies in adults and form the basis of this report. The majority of Canadian gastroenterologists followed published guidelines, with 62% screening patients without dysplasia every two years. Patients with low grade dysplasia were screened more frequently, with 54% of respondents performing endoscopy every six months, and 35% on a yearly basis. Biopsy protocols showed the greatest variation, with 46% of gastroenterologists taking four-quadrant biopsies at 2 cm intervals along the columnar-lined (Barrett's) esophagus. Seventy-six per cent of gastroenterologists agreed that all patients with chronic gastroesophageal reflux should have a "once in a lifetime" endoscopy to screen for Barrett's esophagus. The majority of Canadian gastroenterologists follow current guidelines for the management of Barrett's esophagus and support the concept of "once in a lifetime" endoscopy. PMID- 12772006 TI - 6-mercaptopurine and inflammatory bowel disease: hidden ground for the cytomegalovirus. AB - 6-mercaptopurine (6-MP) and azathioprine are important drugs for the treatment of inflammatory bowel disease (IBD) but their actions suppress host defense against infection. A challenging case of a 19-year-old female patient with quiescent Crohn's disease maintained with 6-MP presenting with dyspnea and a normal chest exam and x-ray is presented. She became ventilator-dependent and only after numerous investigations was diagnosed with cytomegalovirus (CMV) pneumonitis. A systematic literature review of CMV infections in IBD patients was performed. The present case is the first report of a patient with quiescent IBD maintained on 6 MP who developed CMV pneumonitis. Other reports have identified patients with active disease on multiple immunosuppressants who developed CMV pneumonitis and also highlight the risk of CMV colitis in refractory IBD. The authors review the approach to the diagnosis of CMV infections in IBD patients with atypical pneumonia and colitis and highlight the importance of considering CMV infection in these settings. PMID- 12772007 TI - Prognostic indicators in acute pancreatitis. AB - Several approaches have been used in an attempt to predict the severity and prognosis of attacks of acute pancreatitis. The Ranson and Glasgow criteria include a variety of simple laboratory parameters that are measured on admission and again within 48 h. They are the most widely used indices in clinical practice. The Acute Physiological and Chronic Health Evaluation II system is more complicated, but can be applied to a wide variety of conditions, especially in intensive care settings. The usefulness of this system depends on the threshold score for defining severe pancreatitis; a score of eight appears to be the most appropriate. The finding of nonperfused areas in the pancreas at contrast enhanced computed tomography is indicative of pancreatic necrosis and portends an unfavourable prognosis. Other clinical and laboratory indices have been proposed, but the most important predictive factor of early mortality seems to be the presence and persistence of a Marshall organ failure score of two or more. This is especially true if organ dysfunction persists beyond 36 h. Radiological findings do not always correlate well with the presence of organ dysfunction, and more investigations are required. PMID- 12772008 TI - Management of anticoagulants before and after endoscopy. AB - The risk of procedure-related bleeding while taking anticoagulants needs to be weighed against the risk of thromboembolism from discontinuing these drugs. It is not necessary to adjust anticoagulation for low-risk procedures, such as upper endoscopy with biopsy, colonoscopy with biopsy or endoscopic retrograde cholangiopancreatography with stent insertion (but without sphincterotomy). Procedures that incur a high risk of bleeding include polypectomy, endoscopic sphincterotomy, laser therapy, mucosal ablation and treatment of varices. For these procedures, warfarin should be discontinued four to five days beforehand. Depending on the risk of thromboembolism, that is based on the nature of the underlying condition, the patient may require vitamin K and/or fresh frozen plasma (to ensure that coagulation parameters are within the normal range) or heparin infusions (to ensure that some degree of anticoagulation is maintained). Low molecular weight heparin is an alternative to unfractionated heparin for select cases with a high risk of thromboembolism. Warfarin therapy may generally be resumed on the night of the procedure and may be supplemented by heparin in patients with a high risk of thromboembolism. It is not necessary to discontinue acetylsalicylic acid or nonsteroidal anti-inflammatory drugs, when used in standard doses, for endoscopic procedures. There are insufficient data to make recommendations regarding newer antiplatelet drugs, such as ticlopidine or clopidogrel, but it is prudent to discontinue these medications seven to 10 days before a high-risk procedure. PMID- 12772009 TI - Motion--Cyclo-oxygenase-2 selective nonsteroidal anti-inflammatory drugs are as safe as placebo for the stomach: arguments against the motion. AB - Cyclo-oxygenase (COX) exists in two isoforms, COX-1 and COX-2, that direct the synthesis of prostaglandins, prostacyclin and thromboxane. Traditional nonsteroidal anti-inflammatory drugs (NSAIDs) inhibit both isoenzymes, resulting in damage to the mucosa of the stomach and duodenum, but also in cardioprotection. Selective COX-2 inhibitors are less likely to damage the upper gastrointestinal tract, as has been shown by large, randomized, controlled trials. Specifically, the newer agents are superior to ibuprofen and naproxen in this regard, but celecoxib and diclofenac were not significantly different in patients who were not also taking low-dose acetylsalicylic acid. These studies did not include a placebo arm, however, and controlled comparisons of COX-2 inhibitors with placebo have not enlisted enough subjects to demonstrate conclusively that they are equally safe. Selectivity for the COX-2 isoform affords protection against upper gastrointestinal toxicity possibly at the expense of the cardioprotective effect of traditional NSAIDs. This might explain the higher rate of nonfatal myocardial infarction in patients who are given rofecoxib compared with naproxen. A traditional NSAID, combined with either misoprostol or a proton pump inhibitor, is still a suitable alternative to selective COX-2 inhibitors for the treatment of arthritis. PMID- 12772011 TI - Activities of the Canadian Association of Gastroenterology Endoscopy Committee. PMID- 12772010 TI - Motion--Cyclo-oxygenase-2 selective nonsteroidal anti-inflammatory drugs are as safe as placebo for the stomach: arguments for the motion. AB - Traditional nonsteroidal anti-inflammatory drugs (NSAIDs) are known to cause gastritis, gastric and duodenal ulcers, and gastrointestinal (GI) blood loss, as well as alterations in small bowel permeability. Patients at a high risk for these complications include those who are older than 60 years of age, those with a previous history of complicated peptic disease and bleeding, and those who take high dose or multiple NSAIDs, including low dose aspirin, corticosteroids or anticoagulants. The introduction of selective inhibitors of cyclo-oxygenase-2 (COX-2) has provided effective treatment of inflammatory arthritis and musculoskeletal pain, with dramatic reductions in the risk of GI adverse events. The two most widely prescribed coxibs are celecoxib and rofecoxib, and others are being developed. Endoscopic studies have revealed that coxibs are only half as likely to induce upper GI ulceration than are traditional NSAIDs, and are as safe as placebo. Furthermore, the newer drugs do not cause excessive blood loss from the GI tract and do not affect small bowel permeability. The Vioxx Gastrointestinal Outcomes Research Study (VIGOR) revealed that the incidence of myocardial infarction was significantly lower with naproxen than rofecoxib, although this study was not designed to look at this endpoint. Coxibs are an important addition to the pharmacotherapy of inflammatory disease. PMID- 12772012 TI - Working together. PMID- 12772013 TI - Quelling the fire. PMID- 12772014 TI - 2001 Canadian Cardiovascular Society Consensus Conference on cardiac transplantation. PMID- 12772015 TI - An overview of the methods and data used in the CCORT Canadian Cardiovascular Atlas project. AB - The Canadian Cardiovascular Atlas project, an initiative of the Canadian Cardiovascular Outcomes Research Team (CCORT), will be published as a series of 20 articles in future issues of the Canadian Journal of Cardiology. Through a wide range of data sources and analyses from a number of collaborators across Canada, the CCORT Atlas will provide a comprehensive overview of the current state of cardiac care and disease in Canada. Administrative data, clinical registries and community survey data will be analyzed at the provincial and health region levels. The purposes of this article are to 1) provide an overview of the data types and sources used in the Atlas project, 2) give a general description of the methods and analyses used to report Atlas data and 3) describe how Atlas maps were created and how they can be interpreted. PMID- 12772016 TI - Rapid increase in statins newly dispensed to Ontario seniors between 1994 and 2000. AB - BACKGROUND: The Ontario government spent approximately 10% (CDN $148 million) of the provincial medication budget on statins in 1998. This number can be expected to grow in light of new guidelines from the United States recommending that three times as many patients should be receiving antihyperlipidemic therapy. There is scant population-based information on the age and sex distributions of patients receiving these medications. OBJECTIVES: To estimate the rates of new use of statin medications among community-dwelling elderly adults in Ontario between 1994 and 2000. METHODS: Data from the Ontario Drug Benefit program were used on all medications dispensed to noninstitutionalized Ontarians over 65 years of age to estimate age- and sex-specific annual rates of patients newly dispensed a statin. Changes in rates were estimated using Poisson regression. RESULTS: The number of elderly Ontarians newly dispensed a statin increased rapidly between 1994 and 2000, with age-standardized rates rising from approximately 840 to 2600/100,000 women and from 810 to about 3100/100,000 men. The highest rates of new use were observed among patients of both sexes aged 65 to 74 years. However, the rate of change increased with advancing age so that the biggest increase was observed among those aged 85 years and above, among whom there was an 8.5-fold increase among women and a 12-fold increase among men. CONCLUSIONS: Statins have been shown to be safe, efficacious and cost effective in reducing the risk of sudden cardiac death and other acute coronary events among middle-aged patients with pre-existing cardiovascular disease. The present study showed that there was a rapid increase in the rate of statins newly dispensed to elderly patients in Ontario, among whom estimates of safety, efficacy and cost effectiveness are not well quantified. Better estimates of these parameters in the elderly are required because of the high costs and benefits and potential unintended beneficial and harmful effects of statins. PMID- 12772017 TI - Intima-media thickness of the common carotid artery is the significant predictor of angiographically proven coronary artery disease. AB - BACKGROUND: Previous studies have shown a relationship between intima-media thickness (IMT) of the common carotid artery and coronary artery disease (CAD). The role of IMT in the prediction of significant CAD has not been established. OBJECTIVES: To investigate the diagnostic accuracy of IMT measurement and the detection of carotid plaques in relation to cardiovascular risk factors in the prediction of significant CAD. PATIENTS AND METHODS: One hundred and seventy patients (121 men and 49 women; average age 58 +/- 11 years) undergoing selective coronary angiography were examined by carotid ultrasound. IMT was measured. Plasma lipid concentrations and other risk factors were determined. RESULTS: Angiographically proven significant CAD was found in 138 (81%) of all patients. Carotid plaques were detected in 98 (58%) of all patients. Presence of carotid plaques in common carotid artery (P<0.001) and male sex (P<0.005) were found to be categorical risk factors for significant CAD but in multiple regression analysis only age (P=0.15), IMT (P<0.01), high density lipoprotein (HDL) cholesterol (P=0.02) and, less significantly, total cholesterol (P=0.09) were found to be independent parameters for the prediction of significant CAD. IMT of 0.75 mm was determined as a cut-off point for the detection of significant CAD (sensitivity 78%, specificity 79%, positive predictive value 95%, negative predictive value 41%, odds ratio 12.9, 95% CI 3.5 to 47.6). CONCLUSION: The increase in IMT is the significant positive predictor of angiographically proven CAD; other predictors are high age, low HDL cholesterol and, less significantly, high total cholesterol. Presence of carotid plaques and male sex do not add any new information for the prediction of CAD once the predictors are considered. PMID- 12772018 TI - Prevention of radial artery graft spasm: a survey of Canadian surgical centres. AB - BACKGROUND: The radial artery (RA) is used as an alternative conduit to the traditional saphenous vein bypass graft in some cardiac surgery centres. Despite limited clinical trial data, many centres appear to be using routine drug therapy to prevent spasm of the RA graft in the immediate and late postoperative period. OBJECTIVES: To determine the proportion of patients in Canadian cardiac surgery centres having RA bypass grafts and the percentage of patients routinely receiving drug therapy to prevent spasm of these grafts. METHODS: A questionnaire was sent to all cardiac surgery centres in Canada requesting data for the year 2000 on the total number of bypass surgery operations and the number of patients having RA bypass grafts. Information on approaches taken toward prophylaxis against spasm of RA grafts was also requested. RESULTS: The use of RA grafts ranged between 1% and 50%. Twenty-five of 27 centres used antispasm prophylaxis with calcium channel blockers with or without long acting nitrates both in the intensive care unit and on the surgical ward. Calcium channel blockers were prescribed routinely in all 25 centres with 12 centres also using long acting nitrates. Only two centres routinely continued nitrates on the ward. The duration of antispasm therapy varied from several weeks up to six months. CONCLUSIONS: Prophylaxis against spasm of RA grafts is practised in over 95% of Canadian surgical centres despite limited clinical outcome data to support this practice. PMID- 12772019 TI - A pilot study of exercise training in adult patients with repaired tetralogy of Fallot. AB - BACKGROUND: Positive effects of physical training in adults with acquired heart disease have been reported. The role of exercise training in adults with congenital heart disease, however, is less well defined. OBJECTIVES: To assess the level of physical activity in adult patients with repaired tetralogy of Fallot (TOF), as well as the safety and effect of exercise training in these patients. PATIENTS AND METHODS: A questionnaire of physical activity was filled upon entering the study by the participants. Eighteen adult patients with repaired TOF were then randomly assigned to participate in a three-month structured exercise program (exercise group, nine patients) or to continue their usual exercise routine (control group, nine patients). Each patient in the exercise group received an individualized exercise program to be performed three times a week, whereas the control group was told to continue living their life as usual. Cardiopulmonary testing was performed in all patients at baseline (before randomization) and at the end of the study. RESULTS: No death or incidents occurred during the study period. All patients but one were physically inactive at the onset of the study. There was a significant increase in peak oxygen consumption in the exercise group by the end of the study (22.1 mL/kg/min [6 metabolic equivalents (METS)] versus 24.3 mL/kg/min [7 METS], P=0.049), whereas it remained unchanged in the control group (21.8 mL/kg/min [6 METS] versus 22.1 mL/kg/min [6 METS], P=0.825). CONCLUSION: In clinically stable adult patients with repaired TOF, a moderate level of exercise training improves aerobic capacity. Regular exercise should be encouraged in these sedentary patients. PMID- 12772020 TI - Effects of radial stretch on target lesion revascularization after percutaneous coronary intervention: an intravascular ultrasound study. AB - OBJECTIVE: To analyze possible associations between radial stretch during coronary angioplasty and the incidence of target lesion revascularization (TLR). PATIENTS AND METHODS: Intravascular ultrasound images were obtained before and after revascularization in 182 native coronary lesions. The quantitative intravascular ultrasound parameters (external elastic membrane cross-sectional area [EEM-A], lumen areas [LA], plaque area [PA], calculated as EEM-A-LA, and changes between pre- and postinterventional LA [DLA], EEM-A [DEEM-A] and PA [DPA]) were correlated with the incidence of TLR. RESULTS: TLR was performed in 60 (33%) patients, while 122 (67%) patients remained event free. Postinterventional PA remained significantly larger in the TLR group than in the event-free group (9.2 +/- 3.3 mm2 versus 7.9 +/- 3.3 mm2; P=0.02). The radial stretch during intervention, expressed as DEEM-A, and the balloon to artery ratio was significantly larger in the TLR group (DEEM-A: 1.9 +/- 2.1 mm2 versus 1.3 +/- 2.0 mm2; P=0.03; balloon to artery ratio: 1.3 +/- 0.2 versus 1.0 +/- 0.5; P=0.04). Multivariate analysis revealed DEEM-A (P=0.01), DPA (P=0.03), diabetes (P=0.001, odds ratio 5.2, 95% CI 4.9 to 6.5) and adaptive remodelling (P<0.001, odds ratio 4.1, 95% CI 3.5 to 6.4) as independent predictors for TLR. CONCLUSION: Whereas patients in whom lumen gain is achieved primarily by plaque reduction with less wall stretch tend to remain event free, patients with significant radial stretch (ie, less reduction of the PA, but a radial outward shift of the plaque mass) experience a higher incidence of TLR. PMID- 12772021 TI - Paced breathing can prevent vasovagal syncope during head-up tilt testing. AB - BACKGROUND: Clinical experience suggests that, through the control of heart rate, paced breathing may prevent syncope. OBJECTIVE: To assess the effect of metronome paced breathing (0.2 Hz) on the outcome of head-up tilt test (HUT) in patients with vasovagal syncope. PATIENTS AND METHODS: Ten patients (two men), mean +/- SD age 18.4 +/- 5.1 years, with a positive HUT with spontaneous breathing and no evidence of cardiovascular or neurological disease, were exposed to a second HUT under metronome-paced breathing (0.2 Hz). Continuous electrocardiographic recordings were obtained during the two tests, and heart rate variability was analyzed off-line. RESULTS: During HUT with metronome-paced breathing, only one of the 10 patients developed bradycardia, hypotension and syncope (P<0.01, Wilcoxon). Compared with HUT with spontaneous breathing, HUT with paced breathing induced a larger increase in the heart rate in the first minute of tilt (4 +/- 6 beats/min versus 16 +/- 12 beats/min, P<0.05). CONCLUSIONS: The results show that paced breathing can prevent vasovagal syncope induced by HUT. This finding suggests that respiratory training could be useful to prevent vasovagal syncope. PMID- 12772022 TI - Primary and secondary prevention of cardiac disease: bridging the two solitudes. AB - Clinical cardiovascular medicine uses two models for atherosclerosis: primary and secondary prevention. The essential goal of both strategies is risk factor modification, but intervention strategies and funding mechanisms to achieve these goals have traditionally differed, leading to professional and clinical segregation. The current system defies biological reality, patients and families are not well served, and important opportunities for improved care are lost. Developing a more inclusive paradigm of primary and secondary prevention remains a significant challenge. Such a paradigm could transform a longstanding trend of two solitudes where efforts in one area rarely benefit from new developments in the other. New models need to be developed that balance and integrate diagnostic, curative and preventive care, and that coordinate public health and personal health services. Working on 'bridging strategies' will help create synergies that will lead the way to further integration and will help improve the heart health of the population. PMID- 12772023 TI - Monckeberg's sling. PMID- 12772024 TI - Chorea in a pregnant woman with rheumatic mitral stenosis. AB - Chorea gravidarum is a rare movement disorder of pregnancy with a broad differential diagnosis. Although often a benign condition, it may indicate underlying acute rheumatic fever, antiphospholipid antibody syndrome or a hypercoagulable state. However, now that rheumatic fever is rare in western countries, chorea gravidarum occurs most commonly in patients with chronic rheumatic heart disease. Heightened awareness of chorea gravidarum and the morbidity of the often associated rheumatic heart disease, particularly in immigrants from developing countries, is essential for early diagnosis and effective management. A case of chorea gravidarum in a woman with rheumatic mitral stenosis is described. The diagnostic approach, pathophysiology and management of this rare condition are discussed. PMID- 12772025 TI - Myocardial infarction in a young woman secondary to a rupture of a noncoronary sinus of Valsalva aneurysm without coronary artery disease. AB - A 22-year-old woman presented with sudden onset of chest pain. Echocardiography showed a ruptured aneurysm of the noncoronary sinus of Valsalva in the right atrium. Slight ST segment depression was observed on initial electrocardiography (ECG). The patient was transferred to a tertiary care centre 10 h after the onset of symptoms. Surgery consisted of patch closure of the noncoronary sinus and tricuspid valve resuspension through the right atrium. Postoperatively, myocardial infarction (MI) was diagnosed based on a significant increase in cardiac enzymes and a new septal and apical akinesia on echocardiography. The etiology of MI in such a setting is multifactorial. Decreased coronary perfusion secondary to the severe aortic valve regurgitation and increased left ventricular end diastolic pressure, coupled with increased myocardial workload and delay before surgery may be implicated in the genesis of MI. PMID- 12772026 TI - Evidence-based decision-making. PMID- 12772027 TI - Occurrence of a secondary primary papillary fibroelastoma. PMID- 12772037 TI - [Madrid Declaration on Ethical Standards for Psychiatric Practice]. PMID- 12772038 TI - [Attitudes towards change in eating disorders (ACTA). Development and psychometric properties]. AB - INTRODUCTION: The aim of the present study was the development of a self-reported instrument in Spanish to assess attitudes towa rds change in eating disord e rs (AC TA ) and to analyze its reliability and validity. METHODS: The questionnaire was elaborated following the transtheoretical approach of stages of changes, proposed by Prochaska and DiClemente and using the clinical records systematically registered regarding patients cognitions, behaviors and emotions related to the disorder. It was administered to 186 patients who where diagnosed an eating disorder according DSM-IV criteria. Subsequently, the process of refinement and validation of the scale was initiated. Moreover, a set of self reported instruments was used to assess the eating disorder psychopathology: the Eating Attitudes Test (EAT), the Bulimic Investigatory Test Edinburgh (BITE), the Eating Disorders Inventory (EDI-2) and the Body Shape Questionnaire (BSQ). RESULTS: The final version consisted of 59 items divided into six subscales: precontemplation, contemplation, determination, action, maintenance and relapse. All of them s h owed an internal consistency over 0.70 which corresponded to the six factor obtained after the factorial analysis. Furthermore, the subscales were logically correlated to each other and to the questionnaires measuring eating psychopathology. CONCLUSIONS: The present results suggest that the ACTA be an easily administered, reliable and valid questionnaire, which could be used withinthe motivational approach. This could provide interesting information regarding the knowledge of the therapeutical process. PMID- 12772039 TI - [GEOPTE Scale of social cognition for psychosis]. AB - INTRODUCTION: Despite the large number of scales to assess cognitive function, these are rarely used in clinical practice, both because of the time they require and because they do not give useful information to the clinician. The aim of this article is to present the characteristics and psychometric properties of a scale which aims, with its simplicity of use and design, to be of use in the clinical practice for measuring social cognition in psychosis. METHODS: The new GEOPTE Scale gathers information from two sources: the patient's subjective perception of his/her deficits and that of the informant or caregiver. It consists of 15 items (7 for basic cognitive functions and 8 for social cognition). The scale was applied to 87 patients with a diagnosis of psychosis (according to DSM-IV), and general clinical data, clinical global impression, mood and degree of insight were gathered. RESULTS: The GEOPTE Scale presented excellent internal consistency (Cronbach's alpha 0.84 for patient and 0.87 for informants). Factorial analysis identified two factors which explained a total variance of 39%. The first factor was related to the basic cognitive function items and the second to the social cognition items. Regarding the validity of the construct, the scores on the scale are closely related to clinical global impression, degree of insight and depressive symptoms. CONCLUSIONS: The GEOPTE Scale for measuring social cognition in psychosis has an excellent psychometric behavior both in the degree of internal consistency and in correlation with clinical global variables, mood and degree of insight. PMID- 12772040 TI - [Detection of bulimia nervosa in primary health care consultations]. AB - INTRODUCTION: To estimate the prevalence of bulimia nervosa and bulimic behavior in women who come to Primary Health Care consultations, using the DSM-IV criteria. DESIGN: Descriptive, observational study Setting. Primary Health Care. PARTICIPANTS: Women over 14 years who come to general medicine consultations. Selection by systematic sampling of 175 women (proportion expected: 4%; confidence index: 95%; accuracy: 3%). MAIN MEASURES: Examination, by semi structured interview, of diagnostic criteria of bulimia (recurrent binge eating, compensatory behaviors and excessive concern about weight or body image), socio demographic variables and variables on morbidity (health problems according to CIPSAP-2-defined). RESULTS: Prevalence of bulimia was 5.3% (95% CI: 2.4-9.7), there being recurrent binge-eating in 23.4%. Among the compensatory behaviors, fasting (13.5%), intense exercise (8.2%) and self-induced vomiting (4.7%) were the most frequent. In women with bulimia, a background of anorexia and establishment of goals to lose weight appeared more frequently (p <0.05). Their average age was significantly lower (p <0.05) (31.2 14.7 SD). By logistic regression, the associated variables with the presence of bulimia were age (OR: 0.94), existence of previous psychiatric morbidity (OR: 9.0) and having previously set goals to lose weight (OR:7.3). CONCLUSIONS: In the women who came to the Primary Health Care consultations, prevalence of bulimia is greater than that described in the general female population. This disorder is more frequent in younger women and in those who present a background of psychiatric morbidity. Examination of the diagnostic criteria makes its detection easy in Primary Health Care consultations. PMID- 12772041 TI - [Study of translation and reliability of the Wisconsin personality disorders inventory (WISPI-IV)]. AB - INTRODUCTION: The Wisconsin personality disorders inventory (WISPI-IV) is a self report instrument for the assessment of DSM-IV personality disorders. The aim of this study was to translate the WISPI-IV into Spanish and report its internal consistency and temporal stability. METHODS: The Spanish version was obtained by translation and back translation method, using two independent translators. Then 270 subjects of both genders, between 18 nd 65 years of age, who knew how to read and write and accepted to participate voluntarily were selected and answered the instrument. A subgroup of 20 subjects completed the instrument for the second time, at fifteen days of the first application. The analysis of internal consistency and coefficients of temporal stability were calculated with Cronbach's alpha coefficients and Pearson's correlation, respectively. RESULTS: All Spanish version WISPI-VI scales mean scores and internal consistency coefficients were similar to their original counterparts (alphas=0.64-0.86). Except for avoidant personality disorder scale, test-retest coefficients were also moderate to high and statistically significant (r = 0.46-0.92). CONCLUSIONS: The Spanish version of the WISPI-IV behaved similarity with the English version, and demonstrated adequate internal consistency and temporal stability coefficients to evaluate the presence of personalisty disorders. PMID- 12772042 TI - [Enhanced suppression of cortisol after dexamethasone in borderline personality disorder. A pilot study]. AB - INTRODUCTION: Some studies have suggested the etiological role of childhood traumatic events in borderline personality disorder (BPD), involving the stress response mechanisms and the activity of hypothalamic-pituitary adrenal (HPA) axis. Recent preliminary results show that BPD, similar to that found in post traumatic stress disorder (PSD), might have a hypersensitive response to the dexamethasone test. METHODS: Fourteen BPD patients, diagnosed according to DSM-IV criteria, without a major depressive episode or history of bipolar or psychotic disorder, were compared with 10 patients with other personality disorders (OPD). Plasma cortisol was measured at baseline and following an oral test with 0.25 mg of dexamethasone. RESULTS: Nine out of 14 (64%) BPD patients were cortisol suppressors in the test versus only 2 out of 10 (20%) patients with other personality disorders (chi square 4.6, degree factors [df] 18, p<0.05). The degree of cortisol suppression was significantly greater for BPD patients (73% ) than for patients with other personality disorder (34 %). Baseline cortisol concentrations, although lower in BPD patients, were not significantly different among groups. CONCLUSIONS: BPD could be associated with hypersensitivity of feedback mechanisms of the HPA axis similar to PSD, which suggests a possible role for traumatic experiences in the pathogenesis of the disorder. PMID- 12772044 TI - [Up-date in the treatment of rapid cycling and other refractory bipolar disorders]. AB - INTRODUCTION: Up-date in the treatment of rapid cycling and other resistant bipolar disorders. METHODS: A Medline research of the literature was performed in several databases as Pub-Med, Cochrane and Embasse, from 1998 to December 2001. We have also reviewed bibliography supplied by different laboratories and several monographies. RESULTS: 30 articles were selected: 11 reviews, 17 openlabeled studies and 2 articles on general recommendations. From the 17 open-labaled studies, 10 were on topiramate (25 to 400 mg/day) as a coadjuvant of another stabilizer. Improvement ranged from 40 to 70%; 4 adding gabapentin to the previous treatment, at the dosage of 60 to 5,600 mg/day, 27 and 92% showed improvement; 1 with mexiletine (200 to 1,200 mg/day) in which 46% were full responders, 15% partial responders and 38 % had no response, 100% response in manic or mixed and 38 % in depressed patients; and 2 with lamotrigine (50 to 500 mg/day) in which 52 to 80 % showed improvement. With risperidone at the dosage of 2-3 mg/day as coadjuvant, improvement was seen in 62 %. Olanzapine had direct short-term antimanic effects, with 49 % improvement in single drug therapy and 57 % as coadjuvant. CONCLUSIONS: More double-blind studies are necessary to assess efficacy in monotherapy or as coadjuvants, in short-term or even in monotherapy, and to compare the different treatments with each other as well as with the conventional treatment. The authors agree in pointing out the efficacy of gabapentin and topiramate associated to another stabilizer, and also of lamotrigine in depressed phases. PMID- 12772043 TI - [Family and psychosocial variables in the choice of university studies]. AB - INTRODUCTION: Family order of the children and family size as well as other psychosocial variables on University of the Basque Country (UBC) students are compared in order to relate these data with the choice of type of university studies. METHODS: As a sample, we studied 6,013 students from the UBC in different careers and courses. Mean age was 20.26 years. The following instruments were used: a sociodemographic questionnaire, the Eysenck Personality Scale and Ylla Alexithymia Scale. All them were validated for our country. RESULTS: The presence of women was greater in all the career studies except for engineering where the proportion of men was greater. The data verify the relationship between being the youngest or intermediate child of a family of three or more children and studying Journalism and Fine Arts and that the first born of families of two or more children are more represented in Engineering. It is interesting that there are fewer only children in Medicine where children of families of three or more, both first-born as well as intermediate, go. These variables, as well as extraversion, neuroticism and alexithymia, were different in the different career studies. It was also observed that the female university students scored higher in the neuroticism scale and that the levels of Alexithymia were higher among the men. Further, relations were found between child birth order and family size and personality, in the sense that the Medical and Odontology careers presented lower scores in Alexithymia while the more technical careers such as Engineering are those that present a higher alexithymia. Medicine and Odontology, followed by Mathematics and Journalism, obtained the highest scores in neuroticism. Engineering students obtain the lowest neuroticism. The most extroverted students are those from Journalism, Chemistry, Economics and Odontology. CONCLUSIONS: The choice of university studies is associated to gender, birth order, family size and personality patterns. Personality variables are related to gender, birth order and family size. PMID- 12772045 TI - [Basic elements of ethnopsychopharmacology]. AB - Ethnopsychopharmacology is the pharmacology area that studies the effect of psychoactive drugs on patients having different ethnic characteristics. Some important factors in the management of these patients are the differences in enzymatic characteristics, cultural practices in relation with medicine and food, and the biases of medical professionals in their relationship with patients from different ethnic group (which is called <>). At present, when immigration has become an important issue in our country, some basic knowledge on this subject seems compulsory. PMID- 12772046 TI - [Familiar history of suicidal behavior]. AB - Suicide is such a serious public health problem that it has been proposed as an indicator of mental health of a society. Self-harm, a behavior related with suicide, is also a public health problem with a prevalence 8 to 15 times higher than suicide. Suicide behavior is the result of different social, cultural, biological and psychopathological factors and affects all the cultures. This clinical case of 4 brothers from a family of 8 siblings seen repeatedly due to suicide attempts make it possible to discuss these factors. The family and social report describes a low economical and cultural level. The family climate is marked by aggressive environment and inappropriate care of the children. Different members of the siblings initiated drug consumption as a teenager. It is interesting to point out the high frequency of suicide attempts in this family and the health resources used as well as the deterioration in the quality of life associated. The elevated weight of the fa m i ly factors in the development of the suicide behavioris observed and offers the opportunity of questioning if whether it is the environmental factors, genetic vulnerability to mental disorder or specific predisposition to suicidal behavior that is transmitted in the family. PMID- 12772047 TI - [Severity in dysmorphophobia: description of two cases]. AB - Many authors consider dysmorphophobia (body dysmorphic disorder) as a disorder within the obsessivecompulsive spectrum. Both the phenomenological features and response to serotonergic drugs support this idea. Nonetheless, it is included in the somatoform disorder section within the main diagnostic classifications. Two cases of dysmorphophobia are presented. Both patients were admitted to the psychiatric unit due to secondary complicacions of the disorder. After reviewing the available literature, we discuss the severity that this condition may reach. Comorbid disorders and other complications such as suicide attempts are often present. In addition patients usually hide their suffering. Furthemore, it is important for clinicians to assess the body dysmorphic disorder carefully and rule it out. PMID- 12772049 TI - Capsule endoscopy versus push enteroscopy in patients with occult gastrointestinal bleeding. AB - BACKGROUND: Wireless capsule endoscopy is a new method enabling non-invasive diagnostic endoscopy of the entire small intestine. In this study we prospectively examined the diagnostic precision of capsule endoscopy compared with push enteroscopy in patients with occult gastrointestinal bleeding. METHODS: Between July 2001 and October 2002 we examined 48 patients with suspected disorders of the small intestine using capsule endoscopy. 33 patients with obscure bleeding (19 men, 14 women, mean age 58 +/- 23 years) were prospectively examined using capsule endoscopy and push enteroscopy. RESULTS: On average, the patients had been suffering from chronic gastrointestinal bleeding for 30 +/- 36 (1-120) months. The lowest haemoglobin level was 6.5 +/- 1.6 g/dl (2.3-9.6) and on average 9 +/- 10 (0-50) blood units were transfused. Each patient underwent 4 +/- 2 (1-10) hospitalisations, with a mean 9 +/- 4 (5-17) diagnostic procedures before capsule endoscopy was used. Definitive bleeding sites were diagnosed by push enteroscopy in 7 patients (angiodysplasia [n = 5], ulcers [n = 1], multiple jejunal diverticula [n = 1]). Capsule endoscopy showed a bleeding source in 25 cases (76 %) (angiodysplasias [n = 15], Meckel's diverticulum [n = 1], ulcers [n = 7], ileum diverticulosis [n = 1], B-cell lymphoma [n = 1]). Push enteroscopy localised an additional bleeding source in comparison with capsule endoscopy (multiple jejunal diverticula) in one patient. Both methods of examination were safe and showed no complications. DISCUSSION: The present study shows that capsule endoscopy had the highest diagnostic yield and was superior to push enteroscopy in patients with chronic gastrointestinal bleeding. By using the capsule at an early stage the subsequent therapeutic procedure could be considerably shortened and diagnostic processes could possibly be optimised. PMID- 12772050 TI - Measurement of gastric emptying by 13C-octanoic acid breath test versus scintigraphy in diabetics. AB - In this prospective study, we compared the assessment of gastric emptying by the 13C-octanoic acid breath test to gastric emptying scintigraphy in diabetics. We also examined the relationship between gastric emptying parameters and gastric symptoms and cardiovascular autonomic function. The 13C-octanoic acid breath test and scintigraphy were performed simultaneously in 24 diabetics with a solid test meal (1 egg, doubly labelled with 91 mg 13C-octanoic acid and 50 MBq 99mTechnetium-Nanocoll, 60 g white bread, 5 g margarine and 150 ml water). At fifteen-minute intervals, breath samples were taken over 4 hours and examined by mass spectrometry. In parallel, scintigraphy was performed for 2 hours at one minute intervals. Using breath test data, gastric emptying half time (t (1/2) ), lag-phase (t lag ) and gastric emptying coefficient (GEC) were calculated. Subsequently, the correlation of these results with the equivalent data from scintigraphy were determined employing a regression method. To detect a cardiovascular autonomic neuropathy, a 24-h ECG recording was performed. The prevalence of gastrointestinal symptoms in our collective was assessed by a standardized questionnaire. There was a highly significant positive correlation of both 13C-octanoic acid breath test t (1/2) and scintigraphic t (1/2) (r = 0.8257; p < 0.0001) and 13C-octanoic acid breath test t lag and scintigraphic t lag (r = 0.6302; p < 0.001). The sensitivity of the 13C-octanoic acid breath test was 1 and the specificity was 0.73. In our study, there was no significant association of cardiovascular and gastrointestinal autonomic neuropathy. Furthermore, there was no significant relationship between the prevalence of gastrointestinal symptoms and gastric emptying disorders. We conclude that the 13C-octanoic acid breath test represents a suitable method to measure disordered gastric emptying in diabetics due to its highly significant positive correlation to scintigraphy and due to its validity. It is not possible to predict diabetic gastroparesis on the basis of other autonomic function disorders or because of dyspeptic symptoms. PMID- 12772051 TI - High interobserver variability in endosonographic staging of upper gastrointestinal cancers. AB - Mostly based on results of experienced examiners, endoscopic ultrasound (EUS) has been reported to be highly accurate for locoregional staging of upper gastrointestinal cancers. However, data on interobserver variability among EUS examiners, depending on their experience levels, is sparse. A study was therefore conducted to analyse well-documented videotapes of EUS examinations of 108 patients with resected cancers of the esophagus (n = 55) or stomach (n = 53) in a strictly blinded fashion by 5 examiners, all of whom were experienced in EUS (more than 300 examinations: n = 3, more than 100 examinations: n = 2). Besides the individual accuracy rates in cancer staging, a kappa-statistic was calculated to check for interobserver variability. Under the conditions described, the staging accuracy of all investigators was lower than that usually achieved under clinical routine conditions. The mean T staging accuracy was 41.1 % +/- 9.4 and 46.9 % +/- 5.4 in gastric and esophageal cancers, respectively. For N-staging the respective values were 47.9 % +/- 5.1 (stomach) and 67.7 % +/- 5.4 (oesophagus). Kappa-values were above 0.4 only in the staging of non-invasive esophagogastric tumours of the N0 and T1-category, corresponding to a fairly good agreement among the five investigators. Differences depending on experience levels could not be consistently found. Hence, it can be concluded that endosonographic cancer staging performed in a blinded manner results in a low accuracy and high interobserver variability even among experienced examiners. PMID- 12772052 TI - [Kava, kavapyrones and toxic liver injury]. AB - Kava extracts are obtained from the rhizoma of the kava shrub (Piper methysticum) and contain various pyrones which are used as herbal anxiolytic remedies for generalized anxiety syndromes of low and intermediate grades. The commonly recommended daily dose of 60-120 mg kavapyrones and the duration of the therapy of up to 3 months should not be increased without consultation of a physician and were not followed by most patients, since herbal drugs are considered by the population not only as effective but also as safe. Whereas kava extracts are well tolerated by most patients and rare side effects are rapidly reversible upon drug discontinuation, there are suspected hepatotoxic reactions reported during the last years in temporal and not necessarily causal association with a therapy with kava extracts. Almost 80 % of the patients took kavapyrones in overdose (maximally 480 mg/d) and/or for a prolonged time of more than 3 months up to 2 years. Additional risks factors include co-medication with up to 5 other chemically defined or herbal drugs with in part potentially hepatotoxic properties as well as a genetic deficiency of the hepatic microsomal cytochrome P450 2D6. Severe clinical courses with liver transplantation and possible fatal outcome occurred in 7 patients with overdose and/or long duration of the therapy with kavapyrones. Preventive measures should therefore include a dose of 120, maximally 210 mg kavapyrones per day for 1 month, maximally 2 months, as well as a prescription by a physician. Laboratory test (ALT and gamma-GT) should be done before and during the therapy, and co-medication and alcohol consumption should be avoided. With these measures the hepatotoxic risks under the treatment with kavapyrones might be minimized which are also available via internet and from abroad with possible severe consequences when taken without medical supervision. PMID- 12772053 TI - [Hypnosis in gastroenterology]. AB - Hypnosis is one of the oldest remedies against physical diseases and mental disorders of mankind. The term hypnosis is used for the description of a technique as well as for the description of an altered state of consciousness which is induced by this technique. Hypnosis is a scientific tool in psychophysiological studies of gastrointestinal functions (secretion, motility, visceral sensitivity) and their processing in the central nervous system. Hypnosis is an empirically validated treatment of the irritable bowel syndrome even refractory to medical treatment which is recommended by international expert groups (Rome II) and the British Society of Gastroenterology. In diagnostic upper gastrointestinal endoscopy the relevance of hypnosis as an alternative of intravenous sedation needs to be clarified. Hypnosis cannot be recommended as an alternative for intravenous analgosedation in painful endoscopic therapeutic procedures of the gastrointestinal tract. PMID- 12772055 TI - [Sporadic acute hepatitis E in Germany: an underdiagnosed phenomenon?]. AB - Hepatitis E virus infection is the leading cause of enterically transmitted hepatitis worldwide. Case reports of hepatitis E in individuals in Germany so far related to travel to endemic areas. A 33-year-old man presented with painless jaundice. By serology and liver biopsy, no cause of hepatitis could be identified. After discharge transaminases were persistently elevated. Serology (IgG and IgM) confirmed acute hepatitis E. The transaminases declined to normal values within six months. Detailed anamnestic questioning revealed no travel to an endemic region or contact with individuals who had visited such areas. In addition to our patient, a total of 34 cases of acute hepatitis E were reported to the Robert-Koch-Institute (German center of disease control) in 2001. In five of them, the disease had obviously been acquired in Germany. These data indicate that community acquired hepatitis E virus infection may occur sporadically in Germany and should be considered as a cause of seronegative (non-A-non-B-non-C) hepatitis. PMID- 12772054 TI - The transjugular intrahepatic portosystemic stent-shunt (TIPS) as rescue therapy for complete Budd-Chiari syndrome and portal vein thrombosis. AB - We present a 40-year-old female patient with epigastric pain, ascites, and progressive liver failure, caused by Budd-Chiari syndrome (BCS) with thrombotic occlusion of the right and middle hepatic veins. As underlying diseases, essential thrombocythemia and resistance to activated protein C (APC) due to heterozygote factor V Leiden were found. Initial therapy with heparin caused thrombocytopenia (HIT) type II culminating in thrombosis of the last patent left hepatic vein and further deterioration of liver function. The decision against a surgical shunt and liver transplantation by our surgeons on the basis of the risks involved, prompted us to insert a transjugular intrahepatic portosystemic stent-shunt (TIPS). There was no measurable flow signal in the doppler sonography of the portal vein presumably due to thrombosis. A further evaluation with magnetic resonance tomography and angiography was impossible due to movement artefacts. TIPS initially served as a diagnostic tool allowing direct angiography diagnosed thrombosis of the portal vein, the superior mesenteric and the splenic vein respectively. However, insertion of the TIPS shunt and subsequent fragmentation led to an effective hepatic decompression and full recanalisation of the portal vein. In the present case TIPS simultaneously allowed the diagnosis of portal vein thrombosis and served as rescue therapy of complicated Budd-Chiari syndrome. The potential development of HIT type II should be kept in mind when heparin is given, especially to patients with thrombophilia. PMID- 12772056 TI - [Severe exacerbation of chronic hepatitis B during therapy with corticosteroids and lamivudine therapy and successful short-term combination therapy with interferon-beta and interferon-gamma]. AB - Following an acute episode of undifferentiated collagenosis/autoimmune disease with joint pain, a 50-year-old female patient with known long-standing chronic hepatitis B was treated orally with corticosteroids (30 mg prednisolone in decreasing doses over a period of six months). During this treatment, exacerbation of hepatitis B and massive flare-up under simultaneous treatment with lamivudine occurred (GOT 530 U/L, GPT 791 U/L). Serology was positive for HBs-Ag and anti-HBc-lgM. HBV-DNA titer was > 400,000 copies/ml in polymerase chain reaction. Considering the increased risk of reactivation of autoimmune phenomena during a six months therapy with interferon-alpha, an intensive combination therapy with interferon-beta and -gamma (2 weeks: 1 x 3 MIU nIFN-beta Fiblaferon i.v.; 3 weeks: 1 x 3 MIU nIFN-beta i.v. plus 1 x 50 microg rIFN-gamma Imukine) was carried out over five weeks. After two months this resulted in a complete viral and biochemical response. During an observation period of twelve months no reactivation of the autoimmune disease occurred. PMID- 12772057 TI - [The origin of our journal - together with a laudation for Manfred Mayer]. PMID- 12772058 TI - [Comparison of functional results after reconstruction of the thumb]. AB - Between 1980 and 2000, 26 pollicizations of the index finger and 32 transplantations of the second toe were performed for reconstruction of the thumb after traumatic loss. Twenty-three patients with index finger pollicization and 26 patients with second toe transplantation could be reviewed after an average follow-up of 6.4 years. Vascular complications occurred in two toe transplantations and one index finger pollicization. One transplanted toe was lost after vascular complications. Mobility of the interphalangeal joints was reduced after both procedures. Pinch grip was achieved in 20 of 23 patients after pollicization and 20 of 26 patients after second toe transplantation. Sensitivity was better after pollicization. The most common complaints were cold intolerance after both procedures. Fourty of 49 patients would have the operation again, nine patients would refuse thumb reconstruction. PMID- 12772059 TI - [Finger reconstruction by microvascular second toe-to-finger transplantation in patients with traumatic loss of all fingers]. AB - PURPOSE: To assess the utility of second toe-to-finger transplantation with neurovascular reconstruction in patients with loss of all four digits. METHOD: Analysis of 24 toe-to-finger transplantations in 18 patients regarding over-all survival, complications and secondary procedures, sensibility, function, foot symptoms, and patient satisfaction. The original metacarpophalangeal joint was preserved in 50 % of the transplantations. In six patients, a second transplantation was performed. RESULTS: A mean follow-up of 5.8 years was available in 94 % of the patients. 92 % of the transplanted toes survived, in 38 % of the cases complications occurred followed by a secondary procedure. Two point-sensibility was present in 62 % of the transplantations, the largest range of motion of ca. 50 degrees was obtained in the metacarpophalangeal joint. An increase in the range of motion could be achieved by preservation of the original finger joint. The mean extension lag was 37 degrees independent of the preservation of the metacarpophalangeal joint. Foot symptoms were mild in four patients, in two cases severe donor-site-problems were observed. Cold intolerance was present in 47 % of the transferred toes. Overall patient satisfaction was high with 83 % of the patients confirming their decision to undergo operative treatment. CONCLUSION: Second toe-to-finger transplantation is indicated in patients with traumatic loss of all digits. By this method, a great functional gain could be achieved as well as a high level of satisfaction. The rate of complications and possible foot symptoms should be considered. The preservation of the original metacarpophalangeal joint seems to be of importance for the function of the transplanted toe. PMID- 12772060 TI - [Combined palmar and dorsal approach for complex distal radius fractures]. AB - PURPOSE: The purpose of this paper was to describe our management of complex fractures of the distal radius using a combined dorsal and palmar approach and to evaluate the radiological and clinical outcome. PATIENTS AND METHODS: Twenty out of 26 patients with complex fractures of the distal radius, which were operated on from 1996 to 2000 using a combined dorsal and palmar approach, were followed up on an average of 28 months. According to AO fracture classification, there were one A3, two B3, three C1, two C2 and twelve C3 fractures. Six patients had associated injuries. The treatment was standardized: closed reduction on the day of injury; depending on the soft-tissue situation an external fixator or a forearm splint was applied; every patient underwent a computed tomography before the definitive surgical procedure, which was performed between one and 28 days after the injury when the swelling had subsided. According to the fracture patterns, the distal radius was stabilized using double plates (3), a palmar plate (11) or a dorsal plate (4), screws (1) or K-wires (1) combined with the initially applied external fixator (6). Follow-up included a radiological and clinical examination. The results were graded with use of the modified Mayo Wrist Score. Disability was assessed using the DASH score. RESULTS: All fractures healed, some with minor malunion. Fourteen of the 20 patients had osteoarthrotic changes. Compared to the opposite wrist, range of motion for flexion/extension was reduced by 25 %, radial/ulnar deviation by 20 % and forearm rotation by 5 %. Grip strength averaged 15 % less than that of the opposite side. Modified Mayo Wrist Score revealed 65 % excellent, 30 % good and 5 % poor results. DASH score averaged 18 points. PMID- 12772061 TI - [The BIAX total wrist prosthesis as an alternative to arthrodesis in degenerative and posttraumatic arthritis--early results in twenty-one patients]. AB - The BIAX total wrist prosthesis was first implanted 1983 by Beckenbaugh in the United States. From a total of twenty-one BIAX-prostheses between March 2001 and February 2002, we have inserted eleven in patients for degenerative arthrosis, eight for posttraumatic arthrosis and two for rheumatoid arthritis of the wrist. The average age of the patients was 53.0 +/- 10.7 years with a follow-up between eleven and fourteen months on an average of one year. The motion of the operated joints was preserved on average. We have taken pain as the main criterion for the success of the operation and distinguished between pain at rest, in motion and under exertion. All patients showed a very good pain relief of 5 1/2 values from 6 1/2 before to one after surgery on a scale from one to ten. Pain relief is less under exertion, thus we are reserved with heavy workers of the implantation of the prosthesis. Apart from that, the indication corresponds to the arthrodesis of the wrist. The satisfaction of patients with the prosthesis is very high: 17 patients declared it was much better, three declared it was better and only one patient said it was unchanged. Complications occurred in six patients. The main complication was the postoperative dislocation of the prosthesis in three patients. After repositioning, the joints were long-term stable. The straight secured surgical technique is described in detail. For the majority of patients with posttraumatic or degenerative arthrosis, as well as with rheumatoid arthritis, the BIAX wrist prosthesis is recommended as a first-time operation as an alternative to wrist arthrodesis due to pain relief with preservation of joint motion and good acceptance among patients. PMID- 12772062 TI - [Myosonographic and electromyographic (EMC) evaluation of donor-site morbidity after microsurgical TRAM flap]. AB - Due to improved microsurgical techniques the role of autologous breast reconstruction is expanding. One reason is the decreased donor-site morbidity compared to other techniques. Perforator based flaps (DIEP flaps) seemingly decrease the damage of the abdominal wall due to flap harvesting. However, complication rate in perforator flaps is still considered to be higher than in TRAM flaps. Additionally, operating time is increased due to the meticulous microsurgical dissection of perforators. To evaluate whether donor-site morbidity is caused by harvesting a part of the rectus muscle (as TRAM flaps), 15 patients after unilateral muscle sparing TRAM flap underwent EMG and myosonographic examination of the rectus muscle. EMG is the only technique to assess the muscle functionally, as the electrical activity of the muscle is recorded, and motor unit recruitment can be shown. EMG examination clearly revealed that harvesting of a muscle sparing TRAM flap did not impair the rectus muscle compared to the intact contralateral muscle. However, motor units were enlarged. Myosonography (in contrast to MRI) detected particular damage of isolated muscle fibres indicating nerve-related muscle changes. Taking all findings together, harvesting of a part of the rectus muscle shows a distinct damage to the muscle. The muscle however still acts functionally. This indicates that problems after harvesting a TRAM flap (bulging, hernias etc.) are not due to the partial removal of muscle, but due to the general dissection of the abdominal wall, as seen in other abdominal operations. PMID- 12772063 TI - Functional latissimus dorsi island pedicle musculocutaneous flap to restore elbow flexion in replantation or revascularisation of above-elbow amputations. AB - Two total and one subtotal above-elbow amputations had replantation or revascularization for their severely damaged upper extremities followed by functional latissimus dorsi island pedicle musculocutaneous flap to restore elbow flexion. The mean follow-up was 68 months (range: 14 to 121 months). At final follow-up examinations, the patients had sufficient range-of-motion of their elbows with good strength. Restoring elbow function eliminates one of the most important limiting factors for above-elbow replantations. Functional latissimus dorsi island pedicle musculocutaneous flap is very reliable, has minimal donor site morbidity and offers a wider choice when deciding about arm replantation in the upper arm region by providing a chance of restoring functions. PMID- 12772064 TI - [Endoscopic carpal tunnel release without iatrogenic complication--a report about 1000 procedures]. AB - This is a report about 1000 endoscopic carpal tunnel release procedures using a single-portal system. Not a single iatrogenic lesion was found out at the follow up check. Under consideration of certain technical details there is no risk of lesions. PMID- 12772066 TI - Haemangioma originating from a tendon sheath as an unusual cause of trigger wrist: case report. PMID- 12772067 TI - [Hans Schaefer: the future of social medicine]. PMID- 12772070 TI - [Precocity and prevention of violence]. PMID- 12772071 TI - [Official health certificates between the legal obligation to professional secrecy and the patient's privacy]. AB - Public health officers have to write a medical certificate for public authorities requesting them to do so. This certificate is to contain all the results of medical examinations required for the respective purpose, thus enabling the recipient authorities to take the decisions incumbent on them, such as those judging fitness for service or entitlement to claim. In case the persons concerned agree, a transmission of data is admissible independent of statutory provisions. However, this should always be oriented towards minimum data transmission. Nevertheless, lack of consent or even an adverse intention of the person concerned cannot serve as a blockade in so far as the public health officer has complied with current criminal and data privacy regulations. PMID- 12772072 TI - [Turkish and German patients of general practitioners--diseases, drug expectations and drug prescriptions]. AB - AIM OF THE STUDY: to assess the diseases and complaints and the concepts for treatment of patients and drug prescriptions relative to their ethnic origin: Turkish immigrants (T) or German citizens (D). METHODS: Questionnaire survey of patients of general practitioners before and after consultation. The survey was conducted separately for Turkish and German patients, involving nine GP's practices for each group. Sample sizes were 253 Turkish and 637 German patients, respectively. Only responses of patients younger than 60 years of age were evaluated (T: 216/G: 357). Reason: Relative to the entire Turkish population in Germany the number of ethnic Turks older than 60 is too small for comparative purposes. RESULTS: The two most frequent reasons for a visit to the doctor by Turkish migrants were pain of varied origin (T: 44 % / G: 21 %; p > 0.001) and colds or diseases of the respiratory tract (T: 41 % / G: 25 %; p > 0.001). Turkish and German patients differ significantly with respect to their mentioning of pain and colds. The concept of treatment of the Turkish patients is compared to German patients more adjusted to medicaments and less to counselling and discussion with the doctor. The physicians' prescribing frequency on the other hand conforms primarily to the disease of the patients and not to their ethnic group. To confirm this, compare the share of drug recipients per disease group: Respiratory tract: T: 79 % / D: 84 %, alimentary system: T: 58 % / D: 60 % and locomotor system: T: 49 % / D: 39 %. Secondly the physicians' prescribing frequency conforms to the patients' expectations of medicaments: Of the patients who expected a prescription T: 79 % and D: 77 % respectively were given a prescription and of those who did not expect a prescription T: 55 % / D: 51 % did receive it. Again the patients' ethnic group had no influence on the prescribing frequency. Significantly more Turkish than German patients (T: 23 % / D: 9 %; p < 0.001) received pain-relieving drugs. This is especially true for Turkish patients with illnesses of the respiratory, alimentary and locomotor system. This is the explanation: More Turkish than German patients name pain as their consulting reason. Again the prescribing frequency does not depend on the ethnic group. It depends on the patient' s pain. CONSEQUENCES: The treatment concept of Turkish patients is more directed to drugs. They request a drug more intensively and are more convinced of the medicaments' effectiveness than German patients are. In spite of this, physicians give a prescription according to the indication of the patient independent of his ethnic origin. The insistence on a prescription is significantly higher in Turkish than in German patients. PMID- 12772073 TI - [Hygienic procedures in operation theatres--guidelines and reality. Data obtained on hygiene control measures by public health service at Frankfurt am Main]. AB - PROBLEM: Good hygienic practice in hospitals and other medical institutions is mandatory to avoid or to minimise nosocomial infections. In Germany, official recommendations of the commission for hospital hygiene and infection control have been published as guidelines for hygiene management in hospitals as well as guidelines for control measures of the public health services who are obliged by law to control the hygienic situation in hospitals. In this paper the degree of implementation of these guidelines in the hospitals is studied in respect of the hygiene management in the operation theatre. MATERIAL AND METHOD: The constructive and functional situation and the personal hygienic management were documented in 22 community and private hospitals in Frankfurt am Main, including 40 operation suites with 111 operation theatres. Data of the layout of the rooms were obtained by questionnaire and the hygienic management was observed by means of standardised checklists. RESULTS: Layout of rooms was not compatible with the recommendations of the Guidelines of 1990 and 2000 in many cases--especially with regard to the demand that doors to the operating theatres be closed. Automatic door closers were often either not available or defective. In some theatre areas correct hand disinfection while entering the suite was not possible for lack of disinfectant distributors at appropriate sites. Recommendations regarding surgical drapes and gowns were correctly observed in most cases, with the exception of changing gowns after having gone to the toilet room. Quite often, omission of hand disinfection after glove removal was observed. A discrepancy in hygienic procedures of nursing service and physicians was seen. In 40 % of the operation suites, powdered gloves were still in use. Though in all but one suite air conditioning systems were available, they were not correctly serviced in many cases. Cleaning and disinfection after single operations and at the end of the operating day were carried out correctly in most cases--except in operation theatres for ophthalmologic operations. Here too, manual preparation and disinfection/sterilisation of surgical instruments was common, and inappropriate use of sterilisers as well as the use of so-called "quick"-sterilisers. CONCLUSIONS: The recommendations of the commission for hospital hygiene and infection prevention are generally accepted as guidelines, although the degree of translation into practice was often unsatisfactory. PMID- 12772074 TI - [Experiences with anthrax emergency measures during 2001 and 2002 in the city of Essen]. AB - After the terrorist attacks of 11 September 2001 suspected cases of anthrax also occurred in Germany. No case could be confirmed. From October 2001 to November 2002 the fire brigade of the city of Essen was called in 110 cases of suspected anthrax contamination. In 78 cases specimens were transported to diagnostic laboratories, in 22 cases persons were transported. Only in the first few days patients with suspected contamination had to stay in hospital and underwent chemoprophylaxis. Cooperation between the fire brigade, the local health authorities and the hygienists of the involved hospitals was very intense. It seems necessary to evaluate all the German experiences with suspected anthrax cases to develop risk estimations for different exposure situations and to develop specific recommendations for decontamination, disinfection and initial therapy. PMID- 12772076 TI - [Health and health care in Vietnam]. AB - The South East Asian state of Vietnam is currently undergoing a transition from a centralised socialism to a so-called socialist market economy strongly promoting the private sector. For the last 17 years economy experienced an impressive growth. If the assumption is true that economic growth is positively correlated with the health status of the population, the strengthened economy of Vietnam must go along with an improved health situation and health care system of this country. The following paper evaluates this assumption. It is demonstrated that there is indeed a strongly positive correlation between health and development in many aspects. However, it becomes obvious that economic growth is definitely accompanied by increasing regional and social disparity challenging the health care policy of Vietnam and her international partners. PMID- 12772075 TI - [10 years of observation by public health offices in Baden-Wurttemberg- assessment of human biomonitoring for mercury due to dental amalgam fillings and other sources]. AB - Since 1992, in Baden-Wurttemberg, ten-year old children have been surveyed in the project "Sentinel Health Departments" to study their exposure to environmental pollutants and possible health effects. In the four study areas 1200 children have been investigated every year initially, since 1996 every second year. The data for mercury in body fluids are reported here. The decrease in the body burden of mercury as a result of the declining usage of dental amalgam fillings, was been verified. In 1992/93, of all the children who had been surveyed, the 95 percentile for the body burden of mercury was 3.1 microg/l and in 2000/01 1.35 microg/l. Also to be discussed is the reason why mercury-based cosmetic ointments seriously exceed the HBM-II-intervention-value. Because of using these ointments, concentrations of mercury in urine up to 1400 microg/l were found. A study within the project "Sentinel Health Departments" compared the concentrations of mercury in the urine of adults with those in blood and salvia. The results support the opinion that mercury in urine is appropriate for estimating the mercury uptake from dental amalgam fillings. It can be assumed that these results reflect the situation in the entire Federal Republic of Germany. The ten years' experience confirms that the concept of the "Sentinel Health Departments" is excellently suited to obtain data relevant for environmental health of children. Environmental health protection and the essential gathering of data for future health observation in Baden-Wurttemberg. PMID- 12772077 TI - [Reflections on the World Congress for Nephrology]. PMID- 12772079 TI - [Acute renal failure and hypertension crisis after a technoparty]. AB - HISTORY: A 22 year old patient had noted progressive flank pain without recognizable urinary abnormalities for five days. INVESTIGATIONS: A urologist had noted increased serum creatine (2.1 mg/dl), hypertension (180/100 mmHg) and microhematuria. A post-renal cause was excluded by excretory urography. An interview revealed that the patient had consumed cocaine on weekends since age 19; the acute episode was preceded by a rave party with consumption of a total of 3 g of street quality cocaine. DIAGNOSIS AND TREATMENT: Because of microhematuria with a suggestive nephritic urinary sediment, the patient underwent renal biopsy. It showed acute tubular necrosis and interstitial edema, but no signs of glomerulonephritis and negative immunohistology. The patient received antihypertensive treatment. This led to rapid reversal of elevated serum creatinine and microhematuria was noted, but hypertension persisted. Currently the patient receives ACE inhibitors. CONCLUSION: Similar to what is seen in the US, cocaine use has to be considered in the differential diagnosis of acute renal failure with hypertension. PMID- 12772078 TI - [Uremic patients--late referral, early death]. AB - BACKGROUND AND OBJECTIVE: A number of reports from various countries document that patients with renal failure who are referred late to renal units, have more complications e. g. lack of vascular access when dialysis has to be started as well as longer hospitalisation and have also a higher risk of early death. No data on these points are available from Germany. PATIENTS AND METHODS: In a retrospective analysis the timing of referral to the nephrologists was studied in two Departments of Medicine, e. g. Heidelberg and Vienna, for all patients who started renal replacement therapy. For patients in Heidelberg the relation between timing of referral and survival on dialysis was analysed using the Kaplan Maier-technique. RESULTS: In Heidelberg 280 patients were analysed, 174 men, 106 women, age 61.8 +/- 14.5 years; 136 diabetic patients (9 type 1). They had been referred from GPs (n = 131), specialists (diabetologists, cardiologists; n = 20), emergency departments (n = 33), other hospitals (n = 90) or other institutions (n = 16). The measured median creatinine clearance at the time of referral was 14 ml/min (5-34). The median interval between referral and start of dialysis was 17 weeks. 137 patients had been referred < 17 weeks and 143 patients > or = 17 weeks prior to the start of dialysis. 97 of the 111 patients referred < or = 4 weeks prior to dialysis and 59 of the 169 patients referred > 4 weeks had to be dialysed with a central catheter. There were clear differences in patient survival. In patients referred < 17 weeks before the start of dialysis, the actuarial risk of death during the first 12 month was 34.2 % compared to 5.5 % (p < 0.0001) in patients referred > or = 17 weeks. Even the mortality in the interval between 12 and 24 months after the start of dialysis was clearly higher (15.3 %) in patients with late compared to early referral (11.4 %). CONCLUSION: Late referral of patients with impaired renal function to renal units causes more frequent problems of vascular access, longer hospitalisation, more medical complications, higher costs and higher mortality. Early referral of patients with renal failure is indispensable to improve dialysis outcomes. PMID- 12772081 TI - [Acute renal failure--an review]. PMID- 12772080 TI - [Gitelman's syndrome: an important differential diagnosis of hypokalemia]. AB - HISTORY AND CLINICAL FINDINGS: A 26-year-old woman presented with fatigue, muscle cramps and weakness. Since the age of 8 years she had moderate hypokalemia of unknown origin that was confirmed on multiple occasions. There was no family history of disease. INVESTIGATIONS: Laboratory tests showed moderate to severe hypokalemia with a serum potassium concentration of 2.7 to 3.0 mmol/l, hypomagnesemia, metabolic alkalosis and pronounced stimulation of the renin angiotensin-aldosterone system. Despite normal serum calcium levels, urinary calcium excretion was below the detection threshold. Increased natriuresis was observed after administration of furosemide, but not after administration of hydrochlorothiazide. This finding pointed to the presence of a non-functional thiazide-sensitive sodium/chloride cotransporter in the distal convoluted tubule, characteristic for Gitelman's syndrome. Genetic analysis confirmed the diagnosis of Gitelman's syndrome and documented two heterozygous mutations in the gene encoding the sodium/chloride cotransporter. TREATMENT AND COURSE: The patient was treated with 160 mmol potassium and 30 mmol magnesium supplementation per day. Serum potassium was normalized and magnesium serum levels increased. Weakness and fatigue improved markedly. CONCLUSION: Gitelman's syndrome is an important differential diagnosis in the evaluation of the normotensive patient with hypokalemia. PMID- 12772082 TI - [Simultaneous pancreas and kidney transplantation--a chance for diabetic patients with renal failure]. PMID- 12772083 TI - [IgA-nephropathy: the most common type of glomerulonephritis]. PMID- 12772084 TI - [Moderate renal failure: a significant cardiovascular risk factor]. PMID- 12772085 TI - [Commentary on the ALLHAT study. Antihypertensive and Lipid-Lowering Treatment to Prevent Heart Attack Trial]. PMID- 12772086 TI - Hot and cold spots of recombination in the human genome: the reason we should find them and how this can be achieved. PMID- 12772087 TI - Mutations in the fumarate hydratase gene cause hereditary leiomyomatosis and renal cell cancer in families in North America. AB - Hereditary leiomyomatosis and renal cell cancer (HLRCC) is an autosomal dominant disorder characterized by smooth-muscle tumors of the skin and uterus and/or renal cancer. Although the identification of germline mutations in the fumarate hydratase (FH) gene in European families supports it as the susceptibility gene for HLRCC, its role in families in North America has not been studied. We screened for germline mutations in FH in 35 families with cutaneous leiomyomas. Sequence analysis revealed mutations in FH in 31 families (89%). Twenty different mutations in FH were identified, of which 18 were novel. Of these 20 mutations, 2 were insertions, 5 were small deletions that caused frameshifts leading to premature truncation of the protein, and 13 were missense mutations. Eleven unrelated families shared a common mutation: R190H. Eighty-one individuals (47 women and 34 men) had cutaneous leiomyomas. Ninety-eight percent (46/47) of women with cutaneous leiomyomas also had uterine leiomyomas. Eighty-nine percent (41/46) of women with cutaneous and uterine leiomyomas had a total hysterectomy, 44% at age < or =30 years. We identified 13 individuals in 5 families with unilateral and solitary renal tumors. Seven individuals from four families had papillary type II renal cell carcinoma, and another individual from one of these families had collecting duct carcinoma of the kidney. The present study shows that mutations in FH are associated with HLRCC in North America. HLRCC is associated with clinically significant uterine fibroids and aggressive renal tumors. The present study also expands the histologic spectrum of renal tumors and FH mutations associated with HLRCC. PMID- 12772088 TI - Genomewide linkage analyses of bipolar disorder: a new sample of 250 pedigrees from the National Institute of Mental Health Genetics Initiative. AB - We conducted genomewide linkage analyses on 1,152 individuals from 250 families segregating for bipolar disorder and related affective illnesses. These pedigrees were ascertained at 10 sites in the United States, through a proband with bipolar I affective disorder and a sibling with bipolar I or schizoaffective disorder, bipolar type. Uniform methods of ascertainment and assessment were used at all sites. A 9-cM screen was performed by use of 391 markers, with an average heterozygosity of 0.76. Multipoint, nonparametric linkage analyses were conducted in affected relative pairs. Additionally, simulation analyses were performed to determine genomewide significance levels for this study. Three hierarchical models of affection were analyzed. Significant evidence for linkage (genomewide P<.05) was found on chromosome 17q, with a peak maximum LOD score of 3.63, at the marker D17S928, and on chromosome 6q, with a peak maximum LOD score of 3.61, near the marker D6S1021. These loci met both standard and simulation-based criteria for genomewide significance. Suggestive evidence of linkage was observed in three other regions (genomewide P<.10), on chromosomes 2p, 3q, and 8q. This study, which is based on the largest linkage sample for bipolar disorder analyzed to date, indicates that several genes contribute to bipolar disorder. PMID- 12772089 TI - Crossover interference in humans. AB - Crossing-over between homologous chromosomes facilitates proper disjunction of chromosomes during meiosis I. In many organisms, gene functions that are essential to crossing-over also facilitate the intimate chromosome pairing called "synapsis." Many organisms--including budding yeast, humans, zebrafish, Drosophila, and Arabidopsis--regulate the distribution of crossovers, so that, most of the time, each chromosome bundle gets at least one crossover while the mean number of crossovers per chromosome remains modest. This regulation is obtained through crossover interference. Recent evidence suggests that the organisms that use recombination functions to achieve synapsis have two classes of crossovers, only one of which is subject to interference. We statistically test this two-pathway hypothesis in the CEPH data and find evidence to support the two-pathway hypothesis in humans. PMID- 12772091 TI - Reference accuracy in the Journal of Hand Surgery. AB - PURPOSE: An accurate listing of cited references in journal articles is important for the reader. Many studies have revealed errors in the references cited in a variety of subspecialty journals. We believe that a similar amount of errors, relative to other subspecialty journals, exists in the Journal of Hand Surgery. METHODS: One hundred randomly selected references from each of the 1985 and 1995 Journal of Hand Surgery volumes were analyzed for accuracy of the journal name, title, author name(s), year, volume, and page number(s). References were considered without error if they matched the original article exactly. RESULTS: In 1985, 30% of the references examined contained one or more errors as compared with 11% in 1995. CONCLUSIONS: These error rates are similar to those found in studies of other biomedical journals. Perhaps the decrease in the quantity of errors present in 1995 versus that of 1985 is related to the implementation of a more stringent, revised set of instructions to submitting authors imposed in 1995. PMID- 12772092 TI - Hematoma and distraction arthroplasty for thumb basal joint osteoarthritis: a prospective, single-surgeon study including outcomes measures. AB - PURPOSE: Many surgeons have abandoned the simple trapeziectomy as a surgical treatment option for thumb basal joint arthritis secondary to reports of postoperative weakness. The thumb metacarpal subsiding into the trapezial void has been proposed as the causative factor. The goal of the present study was to evaluate the results of trapeziectomy and postoperative K-wire immobilization of the thumb metacarpal in a distracted position without the use of ligament reconstruction or tendon interposition. METHODS: Twenty-six thumbs in 26 patients from a single surgeon's practice were entered into a prospective single-arm study for surgical treatment of peritrapezial arthritis. Treatment consisted of piecemeal excision of the entire trapezium and 5 weeks of K-wire immobilization of the first metacarpal in slight distraction and opposition. No ligament reconstruction or tendon interposition was used. Motion, strength, stress radiographs, standardized dexterity tests, and outcomes questionnaires including the Arthritis Impact Measurement Scales 2 (AIMS2) were evaluated before surgery and 6 and 24 months after surgery. RESULTS: At 6 months 19 of 26 patients (73%) reported complete relief of pain and at 24 months 92% were entirely pain free. Range of motion evaluation showed 24 of 26 thumbs adducted fully into the plane of the palm and 25 of 26 opposed to the fifth metacarpal head. Comparisons between preoperative and 24-month postoperative strength measurements showed an average 47% increase in grip strength, 33% increase in key pinch strength, and a 23% increase in tip pinch strength over preoperative values. AIMS2 data showed postoperative improvement in "hand and finger function" and "arthritis pain" scales. CONCLUSIONS: After trapezial excision K-wire immobilization in a slightly overcorrected position without tissue interposition or ligament reconstruction restores a stable, pain-free thumb that has superior strength and motion compared with published reports of the more complicated interventions. PMID- 12772093 TI - Trapezial space height measurement after trapeziectomy: a comparison of the use of standard and stress radiographs. AB - PURPOSE: To assess whether the behavior of the trapezial space under stress after excision of the trapezium and insertion of a K-wire to hold the thumb metacarpal base away from the distal pole of the scaphoid for 4 weeks is improved by either palmaris longus interposition or ligament reconstruction (LRTI). METHODS: The trapezial space height and trapezial space ratios were assessed on 50 pairs of standard and stress views obtained 1 year after trapeziectomy. Seventeen of the 50 thumbs had simple trapeziectomy, 15 had trapeziectomy plus LRTI and 18 had trapeziectomy with palmaris longus tendon interposition. The reproducibility of these measurements also was assessed. RESULTS: The 95% limits of agreement for intra- and interobserver differences in the measurement of the trapezial space height were -1 to +1 mm and -2 to +1 mm, respectively. The mean differences between the trapezial heights on the standard and stress radiographs were 1.6 mm (SD, 2.5) for simple trapeziectomy, 1.2 mm (SD, 3.1) for trapeziectomy and LRTI, and 1.2 mm (SD, 2.4) for trapeziectomy with interposition of palmaris longus. The differences between the trapezial space height and trapezial space ratios on the standard and stress radiographs after the 3 different surgeries were not notably different. CONCLUSIONS: Neither the creation of a suspensory ligament nor palmaris longus tendon interposition, as opposed to the isolated placement of a K wire across the trapezial void for 4 weeks, alters the behavior of the trapezial space under stress at the 1-year follow-up examination. It is thus appropriate to use standard radiographs at the 1-year follow-up examination to assess and compare the trapezial space after these different techniques of trapeziectomy. PMID- 12772094 TI - Scaphotrapeziotrapezoid arthrodesis: a follow-up study. AB - PURPOSE: The purpose of this study was to review the results of 800 scaphotrapeziotrapezoid fusions performed over a 27-year period. METHODS: The patients were evaluated for postoperative range of motion, grip and pinch strength, pain, return to work, arthritis, and complications. RESULTS: The overall range of motion was 70% to 80% of the nonoperated side and strength was 69% to 89%. Of the patients 88% returned to previous employment. Arthritis developed in 1.8% of the patients. The overall complication rate was 13.4%. CONCLUSIONS: Scaphotrapeziotrapezoid fusion is a viable treatment option especially for a high-functional-demand young population with significant wrist pathology. Complications, specifically nonunion, can be minimized with careful technique. PMID- 12772095 TI - Total wrist arthroplasty with Destot prostheses in patients with posttraumatic arthritis. AB - PURPOSE: To analyze the functional and radiologic results after Destot arthroplasty, a wrist prosthesis designed for posttraumatic arthritis, and to define the indications for the use of the implant. METHODS: Using the Meuli point score system, 28 Destot total-wrist arthroplasties in 25 patients with stage 2 or 3 scaphoid nonunion advanced collapse and scapholunate advanced collapse were evaluated for 12 to 96 months after surgery. RESULTS: The overall ratings of the study group were excellent in 17 cases, good in 6, fair in 1, and poor in 4. Eighty-four percent showed improved range of motion and grip strength. Four patients experienced postoperative complications. No imbalance or dislocation was noted after surgery. CONCLUSIONS: The Destot implant seems to be a good solution to restore functional range of motion after posttraumatic wrist arthritis when arthrodesis is required by nonmanual laborers older than 50 years of age. PMID- 12772096 TI - The "clenched pencil" view: a modified clenched fist scapholunate stress view. AB - Traditional radiographic imaging of the scapholunate interval is complicated by the lack of a view that combines the dynamism of the clenched-fist stress view with the ability to reliably duplicate the precise angle of pronation that shows optimally the scapholunate interval. We describe a simple radiographic technique that combines these 2 criteria that we have found helpful in the office diagnosis of scapholunate dissociation. PMID- 12772098 TI - Treatment of Madelung's deformity in adults by ulna reduction osteotomy. AB - PURPOSE: Adult patients with Madelung's deformity may present with ulnar-sided wrist pain. Treatment often involves addressing the distal radial deformity. If there is focal wrist pathology and a positive ulnar variance, however, then an isolated ulnar-shortening osteotomy may provide symptomatic relief in these patients. The purpose of this study was to report our results of ulnar-shortening osteotomy without radial osteotomy in adult patients with Madulung's deformity. METHODS: From 1988 to 2001 9 wrists in 9 adult patients with Madelung's deformity and ulnar-sided wrist pain underwent ulnar-shortening osteotomy. The distal radius abnormality was not addressed. All of the patients were women and the average age at the time of surgery was 34 years (range, 29-45 y). Two of the individuals were mesomelic dwarfs and the remaining 7 patients were otherwise normal. Surgery was performed after the patients failed at least 6 months of nonsurgical management. RESULTS: All patients had improvement of their symptoms at an average follow-up evaluation of 42 months (range, 6-112 mo). All of the osteotomies united. One patient required replating for a delayed union. There were no infections and no ulnar carpal subluxation. Ulnar-positive variance correction averaged 4.4 mm. Postoperative range of motion and grip strength were equivalent to the contralateral wrist. CONCLUSIONS: Ulnar-shortening osteotomy is a safe and reliable surgical procedure that can relieve ulnar-sided wrist pain in adult patients with symptomatic Madelung's deformity and positive ulnar variance. PMID- 12772099 TI - Longitudinal radiographic analysis of rheumatoid arthritis in the hand and wrist. AB - PURPOSE: To assess the longitudinal radiographic osseous changes of the wrist and hand other than interphalangeal joints in rheumatoid disease. METHODS: Serial wrist and hand x-rays in 96 patients with long-standing rheumatoid disease were reviewed. The average number of years between initial and most recent x-rays was 15.1. The Larsen scoring system was used to assess the degree and severity of joint involvement. We identified patterns of involvement in the wrist, thumb, and finger metacarpophalangeal (MCP) joints. RESULTS: The radioscaphoid and radiolunate joints had the earliest and most severe progression of all joints studied. Scaphoid erosions often were seen early (27%) and their presence was a predictor of progressive involvement. Ulnar styloid erosions commonly were seen as early isolated findings (25%). The distal radioulnar joint showed a rapid increase in Larsen score and was involved in 78% of patients on late x-rays. The thumb showed considerable late MCP joint disease that often led to boutonniere deformity and the trapeziometacarpal joint had the least rate of progression of all joints studied. The most severely and frequently involved MCP joints were the radial (index and middle), which also had the greatest increase in score over the span of the study. Finger MCP joint disease was observed to progress temporally in a predictable pattern: first radial MCP joints of the dominant hand, followed by the nondominant radial MCP joints, and last the ulnar MCP joints of the nondominant hand with small finger involvement preceding that of the ring finger. Of all MCP joints, the ring finger was least affected. CONCLUSIONS: This study clarified the longitudinal osseous radiographic changes of the wrist and hand (excluding interphalangeal joint) in rheumatoid disease. PMID- 12772100 TI - The efficacy of silver nitrate cauterization for pyogenic granuloma of the hand. AB - PURPOSE: We report a series of pyogenic granulomas of the hand treated effectively in the office with silver nitrate cauterization. METHODS: Twelve patients with 13 pyogenic granulomas of the hand were treated. The average duration of symptoms was 2 months. Masses were removed bluntly, pressure was held proximal to the lesion to effect hemostasis, and the base of the lesion was cauterized with silver nitrate applicators. After treatment patients were instructed to keep the lesions absolutely dry for 2 weeks. RESULTS: Complete resolution of the pyogenic granuloma was achieved in 85% of patients after an average of 1.6 treatments (range, 1-3 treatments). CONCLUSIONS: Pyogenic granuloma of the hand can be treated simply and effectively with silver nitrate cauterization in the office setting. PMID- 12772101 TI - Transition from diffuse cellular infiltration to extensive nodular granuloma as a manifestation of isolated sarcoidosis in the hand: a case report. AB - A patient with soft-tissue sarcoidosis of the hand is described. She initially presented with diffuse soft-tissue infiltration in the digits. After a 12-year time period without steroid treatment the patient developed extensive, tumor-like nodular masses involving multiple digits with lesions extending to the fingertips. Variable clinical features of the soft-tissue sarcoidosis of the hand have been reported as distinctive, independent manifestations of the disease. A transition from the diffuse cellular infiltration to the nodular, tumor-like lesions in the present case shows that the isolated soft-tissue sarcoidosis of the hand may exhibit variable clinical features in a single case. PMID- 12772102 TI - Long-term results of free vascularized second toe joint transfers to finger proximal interphalangeal joints. AB - PURPOSE: To review the long-term clinical results of free vascularized second toe joint transfers for severely damaged finger proximal interphalangeal (PIP) joints. METHODS: Eleven joints in ten patients (1 woman, 9 men) were reviewed between 10 and 22 years after surgery. The average patient age at the time of the surgery was 32 years. Patients were evaluated at a mean follow-up time of 15 years; evaluation included range of motion of the transferred PIP joints and the remaining healthy second toe PIP joints, grip strength, finger and toe pain, finger function, gait disturbance, patient satisfaction, and x-ray changes. RESULTS: The mean active range of motion was 47 degrees that lacks 41 degrees extension and flexes to 88 degrees. The mean grip strength was more than 80% that of the nonaffected hand, and no finger pain or gait disturbances were reported. All patients had some extension lag and flexion contractures but most were satisfied with the clinical and functional results of surgery. In all joints, the joint spaces were preserved, although 1 patient had arthritis and 3 had osteophytes. CONCLUSIONS: It is important to prevent extension lag and flexion contracture to get better results from free vascularized second toe joint transfers. The transferred toe PIP joint is durable. PMID- 12772103 TI - Treatment of chronic, traumatic hyperextension deformities of the proximal interphalangeal joint with flexor digitorum superficialis tenodesis. AB - PURPOSE: To our knowledge, there are no reports in the literature regarding treatment of chronic, posttraumatic proximal interphalangeal (PIP) joint hyperextension deformities with flexor digitorum superficialis tenodesis. The purpose of this study was to describe the surgical treatment and results of flexor digitorum superficialis tenodesis for the treatment of chronic, posttraumatic PIP joint hyperextension deformities. METHODS: Twelve patients were reviewed retrospectively and re-examined at a mean follow-up period of 35 months (range, 6-108 mo). Evaluation included completion of a Disabilities of the Arm, Shoulder, and Hand questionnaire and range of motion (ROM) measurements. RESULTS: There were 5 excellent, 5 good, and 2 fair results. Five patients had a residual flexion contracture at the PIP joint of 5 degrees to 15 degrees, although this did not create any functional impairment as determined by responses to the Disabilities of the Arm, Shoulder, and Hand questionnaire at follow-up evaluation. The 2 patients with fair results had postoperative PIP flexion contractures of 30 degrees and 60 degrees. All 12 patients returned to their previous occupations and recreational activities. CONCLUSIONS: Flexor digitorum superficialis tenodesis is an effective method with predictable results for the treatment of chronic, traumatic hyperextension deformities of the PIP joint. PMID- 12772104 TI - Extension block pinning for large mallet fractures. AB - PURPOSE: The purpose of this report is to review the results of displaced mallet fractures treated with an extension block pin and transarticular fixation of the distal interphalangeal joint. METHODS: We retrospectively reviewed 23 patients with 24 fractures to determine the results of treatment, time to union, range of motion, and associated complications. RESULTS: The average patient age was 24 years and the average fracture size was 40% of the joint surface. Ten patients were treated acutely (less than 10 days), 10 subacutely (10-30 days), and 3 chronically (greater than 30 days). Average time to fracture union was 35 days. At 1-year or greater follow-up evaluation the average extension loss was 4 degrees and the average flexion was 77 degrees. There were no major complications and there were 5 minor complications. Using the established outcome criteria for mallet injuries, 92% had excellent or good results. CONCLUSIONS: The results of this study showed that this surgical technique resulted in rapid fracture union with only minor complications and has excellent functional outcome based on established criteria. PMID- 12772105 TI - The pulp ring avulsion lesion as a new indication for the free dorsal middle phalangeal finger flap: a case report. AB - An ideal reconstruction of fingertip injuries should provide good sensibility and no pain in the activities of daily life. We report a case of a professional trumpet player who sustained a severe avulsion injury when his right index finger was crushed in a slamming door. The soft tissue of the distal finger was debrided circumferentially. To cover the defect we performed a free dorsal middle phalangeal finger flap from the ipsilateral ring finger. This flap provides excellent sensory recovery and an aesthetic outcome. PMID- 12772106 TI - The effect of increased peripheral suture purchase on the strength of flexor tendon repairs. AB - PURPOSE: Previous studies have hypothesized unequal load sharing between peripheral and core sutures in flexor tendon repairs. Most commonly peripheral sutures are placed very near the repair site and characteristically fail before the core strands. We hypothesized that placement of the peripheral sutures farther from the repair site would better optimize load sharing and resist suture pullout, yielding a stronger overall repair. METHODS: To test the hypothesis we developed a mathematical model of the load sharing between core and peripheral sutures. By using this model we predicted that placement of peripheral sutures 2 mm from the repair site would optimize the balance of load between core and peripheral sutures. We then divided and repaired 27 flexor digitorum profundus tendons in 6 ways (core plus peripheral or peripheral sutures only at 1 mm, 2 mm, or 3 mm from the repair site). Tendons were clamped to a custom-built linear loading machine and distracted to failure. RESULTS: There was a clinically and statistically significant increase in strength with an increased distance of the peripheral suture from the repair site showing that core sutures augmented by a 2 mm peripheral repair were stronger than those performed with 1-mm peripheral repairs (50.8 vs 37.1 N). CONCLUSIONS: A peripheral stitch placement approximately 2 mm from the repair site represents a simple modification that can significantly increase the ultimate strength of flexor tendon repairs. PMID- 12772107 TI - Two-portal repair of canine flexor tendon insertion site injuries: histologic and immunohistochemical characterization of healing during the early postoperative period. AB - PURPOSE: In vivo animal studies have indicated that the complex structure of the tendon-bone interface may not be restored after repair even under optimal conditions. Controversy exists about the histologic findings in the early postoperative period after tendon reattachment to bone; this may have impact on biomechanical properties. The objective was to study the histologic structure and immunohistochemical staining of the tendon-bone interface in a large model of digital flexor tendon-bone repair. The hypothesis was that the tendon-bone interface matures and assumes a progressively more anatomic histologic and immunohistochemical appearance during the first 6 weeks after repair. METHODS: Twenty-four canine flexor digitorum profundus tendons were released from their insertion by sharp dissection and repaired to bone. The forelimb was immobilized after surgery and 10 minutes of daily passive motion rehabilitation was performed. Dogs were killed at 10, 21, and 42 days after surgery. Hematoxylin eosin and immunohistochemical staining for types I, II, and II collagen were performed. RESULTS: Although at both 10 and 21 days after surgery substantial inflammation was seen at the tendon-bone repair site, this had decreased markedly by 42 days. Although direct apposition of tendon to bone was seen at 42 days, the mature tendon-bone insertion site was not recreated by this time. Staining for types I and III collagen was diffuse throughout the tendon-bone insertion throughout the interval examined. CONCLUSION: These findings suggest that at 6 weeks after surgery the intact tendon-bone repair site shows minimal histologic and molecular similarity when compared with unoperated specimens. PMID- 12772108 TI - Endoscopic versus open carpal tunnel release: a randomized trial. AB - PURPOSE: This study compared the outcomes in patients assigned to either endoscopic carpal tunnel release (ECTR) or traditional open carpal tunnel release (OCTR). METHODS: An unbalanced randomized clinical trial (91 endoscopic, 32 open) was conducted. Short-term and long-term outcomes were evaluated by a blinded assessor. The primary outcome measures were symptom severity measured on a self report scale and nerve/vascular complications. Secondary outcomes included the McGill pain questionnaire, grip strength, pinch strength, sensory threshold (NK PSSD device, NK Biotechnical Corp, Minneapolis, MN), and time to return to work. RESULTS: Both groups improved on all outcomes. No differences were observed in primary outcomes between the groups at either baseline or follow-up at 1 week, 6 weeks, or 12 weeks after surgery. No significant complications occurred in either group. Grip strength and pain were significantly better at 1 and 6 weeks in the endoscopic group although differences dissipated by 12 weeks. No significant differences occurred in other secondary outcomes. Long-term satisfaction was lower in the endoscopic group, attributable to a 5% rate of re-operation. Lower rates of endoscopic release have occurred at our center once these results were available to surgeons and patients. CONCLUSIONS: No substantive difference in benefit was shown for these 2 methods of carpal tunnel release. PMID- 12772109 TI - The acute effect of peripheral nerve transection on digital thermoregulatory function. AB - PURPOSE: This study tests the hypothesis that major peripheral nerves serve as important routes for adrenergic neural fibers and therefore peripheral nerve injury affects cutaneous perfusion within the nerve's sensory innervation territory. The specific aim of the study was to determine whether an acute isolated peripheral nerve injury would result in alteration of blood flow to a specific digit, digital pain, and/or cold sensitivity. METHODS: The influence of peripheral nerves on their dominant area of autonomic vasomotor function was evaluated in 9 patients who had primary neurorrhaphy after a single complete median or ulnar nerve transection. At a mean follow-up of 47 days the response of digital perfusion to a cold stress was examined in these patients by using isolated cold stress testing and laser Doppler fluxmetry. Patients also completed subjective questionnaires to evaluate their symptoms. RESULTS: The affected hands showed a decreased cutaneous perfusion when compared with the contralateral hands during all 3 phases of the cold stress test. In addition cutaneous perfusion patterns evaluated by laser Doppler perfusion imaging showed markedly decreased cutaneous perfusion patterns in the digits within the innervation territory of the injured nerve. There was no notable difference in digital baseline temperatures and temperature changes after an isolated cold stress test between the most affected digits of the injured hands and corresponding digits of the unaffected, contralateral hands. All 9 patients reported mild to severe digital numbness, 8 reported mild to severe pain, and 4 reported mild to moderate cold sensitivity of the upper extremity. CONCLUSIONS: This study documents that cutaneous perfusion and thermoregulatory function is altered after isolated peripheral nerve injury. PMID- 12772110 TI - Upper limb surgery for tetraplegia: a 10-year re-review of hand function. AB - PURPOSE: To perform a 10-year re-review of hand function outcome for 24 tetraplegic persons who had received bilateral tendon transfers and tenodeses. METHODS: The Lamb and Chan questionnaire with additional questions, the Quadriplegic Index of Function (QIF), the Swanson sphygmomanometer technique for hook grip, the Preston Pinch Meter (PP) for key pinch, and a digital analyzer (DA) for both hook and key pinch were the test instruments used. The QIF and DA had not been used previously. RESULTS: Levels of functional independence and expectations were maintained. Mean hook grip values were maintained for the right hand but increased significantly for the left to reach right hand values. Mean pinch grip values decreased significantly. DA measurements confirmed similar hook grip values for both hands but key pinch values were significantly higher than the PP values. Active transfers averaged approximately twice the strength of tenodeses. CONCLUSIONS: Hand function improvements gained from tendon transfers and tenodeses are maintained over time. PMID- 12772111 TI - Distal biceps tendon repair: comparison of surgical techniques. AB - PURPOSE: Various surgical repair techniques for distal biceps tendon ruptures have been reported, however, the optimal technique is unknown. METHODS: Over a 4 year period 19 distal biceps tendon ruptures were repaired: 9 using a single anterior incision and 10 using a modified 2-incision Boyd and Anderson technique. The patients were followed-up prospectively and independently reviewed. RESULTS: Patient-rated elbow evaluation and Short Form-36 (SF-36) scores improved with time independent of surgical technique. At 1 year the 1-incision group regained more flexion (142.8 degrees vs 131.1 degrees ) than the 2-incision group. There was no difference between groups in supination motion, supination strength, or flexion strength, although recovery of flexion strength was initially more rapid for the 2-incision group. Complications were encountered in 44% of cases treated with a 1-incision technique and in 10% of cases treated with the 2-incision technique; however, most of these were minor transient paresthesias. CONCLUSIONS: The differences between the 2 groups were relatively minor with the Morrey 2 incision technique showing a slightly more rapid recovery of flexion strength and fewer complications as compared with the 1-incision technique. PMID- 12772112 TI - Biochemical composition and histologic structure of the forearm interosseous membrane. AB - PURPOSE: The purpose of this study was to determine the structure and composition of the forearm interosseous membrane (IOM). METHODS: The IOM of 12 cadaver forearms was fixed in formalin. After fixation 5 individual IOM fiber bundles per arm were separated by dissection, excised, and processed with hematoxylin-eosin, trichrome, and Verhoff-vanGeison stains. Nine additional fresh forearms were dissected and 5 IOM fiber bundles per arm were analyzed using the hydroxyproline assay. Bundles were evaluated at ulnar, central, and radial locations. RESULTS: Histologic analysis of the IOM bundles obtained from the 12 fixed forearms showed an abundance of collagen in the main bundle central location (84% +/- 7.8%). A progressive increase in collagen was noted from distal to proximal bundles (r =.72). The hydroxyproline assay of collagen content of the main IOM bundle's central location from the 9 additional fresh forearms was 99.3% +/- 16.5%. There was no difference between bundles or location (power = 0.25 and 0.46). CONCLUSIONS: We found that the IOM possesses a large collagen content arranged in fibrillar structures surrounded by elastin. Collagen was abundant in the proximal bundles and decreased in the distal bundles. PMID- 12772113 TI - Upper-extremity sarcomas in the United States: analysis of the surveillance, epidemiology, and end results database, 1973-1998. AB - PURPOSE: Upper-extremity soft-tissue sarcomas are a rare disease with unclear epidemiology and evolving treatment strategies. Our purpose is to evaluate the incidence of upper-extremity soft-tissue sarcomas and the use of adjuvant radiotherapy in this patient population. METHODS: Using the Surveillance, Epidemiology, and End Results (SEER) database, a national population-based database, we identified all cases of primary upper-extremity sarcoma reported to the 9 national SEER registries in the Untied States from 1973 to 1998. RESULTS: Of the 1,286 upper-extremity soft-tissue sarcomas the average incidence rate is 2.2 cases/million/y, which has not changed significantly from 1973 to 1998. Caucasians' average incidence rate is significantly higher than African Americans' (incidence rate ratio [IRR] = 1.3, p =.02); and men are at a significantly higher risk compared with women (IRR = 1.3, p <.01). The use of adjuvant radiotherapy has increased significantly from 17% in 1973 to 47% in 1998 (p <.01). African-Americans, compared with Caucasians, are significantly less likely to receive adjuvant radiotherapy (odds ratio [OR] =.5, p =.01). CONCLUSIONS: The incidence of upper-extremity sarcomas has not changed significantly over the past 3 decades; however, the disease differentially affects the population with higher rates in Caucasians and men. Based on the results of this study the use of adjuvant radiotherapy is increasing but African Americans are less likely than Caucasians to receive this treatment for upper extremity sarcoma. PMID- 12772114 TI - Treatment of mucous cysts of the fingers: review of 134 cases with minimum 2-year follow-up evaluation. AB - PURPOSE: To evaluate the results of a single surgeon's treatment of mucoid cysts, comparing outcomes between injection and surgery. METHODS: One hundred thirty four cysts were treated, with a minimum 2-year follow-up period. Thirty-one patients had nail ridging or deformity at presentation. Eighty patients had multiple soft-tissue punctures into the cyst with a 25-gauge needle and injection with local anesthetic and steroid. Fifty-four patients had surgical excision and joint debridement. RESULTS: In the injection group, complete resolution of the cyst occurred in 48 cases (60%). Among the 32 that recurred, repeat injections were performed in 8 cases; 3 resolved. No recurrences were noted in the surgery group. Nail ridging resolved after surgery in 25 digits; the remaining 6 digits had partial improvement or persistent ridging. Five infections occurred and were treated successfully with antibiotics (4) or debridement (1), or both. CONCLUSIONS: Aspiration and injection was convenient but had a 40% recurrence rate. Surgery provided definitive treatment with no major long-term problems. PMID- 12772115 TI - Atypical palmar fibromatosis with giant fibrous nodule: a case report. AB - We present an unusual case of palmar fibromatosis. A 62-year-old man had a giant fibrous nodule in the deep fascia of the right palm without any pretendinous cord or flexion deformity at the metacarpophalangeal joint. Primarily soft-tissue tumor and palmar fibromatosis were considered as differential diagnosis of nodular mass but finally the latter was confirmed histologically after resection of the giant nodule. PMID- 12772116 TI - Historic origin of the "Arcade of Struthers". AB - John Struthers wrote in 1848 and 1854 about sites of compression of the median nerve from axilla to elbow. He is best known for describing the rare median nerve entrapment by a ligament from a supracondylar process extending to the medial humeral epicondyle. In 1973, observation of ulnar nerve entrapment associated with a midshaft humeral fracture and subsequent anatomic dissections led to the creation of the term "Arcade of Struthers." Review of Struthers' original writings fails to identify either the use of word "arcade" or description of ulnar nerve compression. Review of published anatomic dissections identifies variations in the origin of the medial head of the triceps, not described by Struthers, that may cause failure of an anterior transposition of the ulnar nerve. Continued use of the term "Arcade of Struthers" is historically incorrect. PMID- 12772117 TI - Renal cell carcinoma with metastases to the triquetrum: case report. PMID- 12772121 TI - Abductor digiti minimi musculocutaneous island flap as an opposition transfer. PMID- 12772119 TI - Anatomy of the ulnar artery as it relates to the transverse carpal ligament. PMID- 12772123 TI - Mechanical thrombectomy: basic principles, current devices, and future directions. PMID- 12772125 TI - Surgical thrombectomy: current role in thromboembolic occlusions. AB - Thromboembolic disease occurs in a heterogeneous group of patients with significant co-morbidities and variable presentations, resulting in high morbidity and mortality rates. To decrease these complications, multiple different endovascular modalities have been developed and used to treat this challenging problem. Physicians are now left with a broad array of endovascular and surgical options. Unfortunately, there are little data that accurately compare these modalities. Therefore, an individual approach to each patient with acute ischemia from thromboembolic disease must be employed. In order to tailor the treatment to the needs of each patient, the physician must be familiar with the techniques, complications, technology, and relative strengths and weaknesses of each treatment modality. This article will first outline the main technical considerations for performing surgical thromboembolectomy. The complications of operative thromboembolectomy will then be discussed followed by the data comparing operative and nonoperative therapy. Finally, clinical scenarios that are best treated by surgical intervention will be described. PMID- 12772124 TI - Adjunctive techniques in percutaneous mechanical thrombectomy. AB - Percutaneous mechanical thrombectomy is an established method in interventional radiology and refers to the removal of acute embolic or thrombotic occlusive material in arteries, veins, or vascular grafts using percutaneous transluminal methods. However, initial complete removal of occlusive material can be achieved only in a minority of patients. The amount of removed material varies with the age and composition of the occlusive material. To achieve sufficient revascularization, adjunctive use of a variety of percutaneous endovascular recanalization techniques is necessitated. Additional treatment with local intra arterial fibrinolysis, balloon angioplasty, stent implantation, endoluminal atherectomy, and other measures results in primary technical success rates of 70% to 100% for revascularization of acutely occluded vessels. The above-mentioned different techniques should not be viewed as competitive treatment modalities, rather a synergistic approach should be offered. The aim of this report is to review different adjunctive techniques in percutaneous mechanical thrombectomy with emphasis on techniques, mechanisms of action, experimental and clinical results, potential complications, and their potential role in view of clinical pathways to treat acute limb ischemia. PMID- 12772126 TI - Peripheral arterial applications of percutaneous mechanical thrombectomy. AB - Various devices have been introduced for the purpose of percutaneous mechanical thrombectomy (PMT). These devices show promise as a valuable treatment option in acute arterial thrombotic occlusions, in addition to the gold-standard surgical method, the Fogarty balloon embolectomy, and local fibrinolysis therapy. Local fibrinolytic therapy cannot be used in the presence of contraindications, and can be time-consuming in limb threatening situations. Surgical intervention can also result in intimal vessel wall injury and is of limited value in infrageniculate occlusions. In this review, currently available PMT devices for peripheral arterial applications will be introduced, and their advantages, drawbacks and finally the reported clinical experience with these devices will be presented. PMID- 12772127 TI - Mechanical thrombectomy devices to treat thrombosed hemodialysis grafts. AB - Treatment of a thrombosed hemodialysis graft is an ideal application for a mechanical thrombectomy device. The soft, acute thrombus contained within the graft can be quickly macerated and removed with minimal risk of damaging native vascular endothelium. There are no universally accepted techniques for using mechanical thrombectomy devices to treat a thrombosed vascular access. To the contrary, there are a variety of thrombectomy methods that are currently utilized by interventional radiologists. Each device has its own niche and is best utilized for certain specific situations. A busy interventionalist should be familiar with several of these thrombectomy devices to most effectively treat the variety of clinical problems that may be encountered. PMID- 12772128 TI - Application of percutaneous mechanical thrombectomy in autogenous fistulae. AB - Articles proving that the majority of thrombosed autogenous fistulae can be recovered by interventional radiology are relatively recent, and the largest series to date have favored mechanical methods including manual catheter-directed aspiration, the Hydrolyzer, Amplatz Thrombectomy device, Arrow-Trerotola device, and the rotating pigtail. Rare contraindications to declotting include infection, fistula immaturity, and large aneurysms. The success rates range from 76% to 100% and compare well with the surgical approach whose effectiveness is not supported by comparable publications. The technical challenges of this outpatient procedure can include difficulties in initial catheterization of the vein or in crossing tight stenosis and actual removal of large thrombi. Although more challenging to declot than grafts, forearm autogenous fistulae are more rewarding. Better long term patency has been achieved in the largest series to date as long as the underlying stenoses are sufficiently dilated (50% 1-year primary and 80% secondary patency rates). The results reported in the upper arm are less good. The unmasking of stenoses in close to 100% of cases warrants stenosis detection programs similar to those for grafts. PMID- 12772129 TI - Mechanical thrombectomy for the treatment of lower extremity deep vein thrombosis. AB - Deep venous thrombosis (DVT) has potentially debilitating long-term sequelae if left untreated. Conventional treatment (systemic anticoagulation with heparin followed by coumadin or low molecular weight heparin) often does not adequately relieves clot burden or symptoms, and patients may be left with post-thrombotic syndrome. Although the advent of catheter-directed thrombolysis has markedly improved the treatment of DVT and long-term outcomes of patients treated for DVT, it remains only partially effective on subacute or chronic clot. Mechanical thrombolysis may work synergistically with catheter-directed thrombolysis to decrease clot burden, treatment time, and complication rates, thereby improving outcomes. PMID- 12772130 TI - Applications of percutaneous mechanical thrombectomy in pulmonary embolism. AB - Percutaneous mechanical thrombectomy (PMT) has matured into a reliable and valuable therapeutic tool in acute vascular diseases. PMT devices are designed to achieve rapid clearance of acute occlusion in large arteries and veins. This article provides a summary of cumulated experience on pulmonary embolism (PE) treatment with PMT devices. PMT devices are a heterogeneous group of devices that uses different forms of energy. Most of the devices do not totally eliminate thrombus rather fragment in small particles. The rationale of PMT is based on the rapid relief of central pulmonary obstruction. PMT in massive PE provides efficacious and safe debulking of centrally located thrombus in PE, lowering pulmonary artery pressures and improving hemodynamics and blood oxygenation. This results in lowering mortality if compared with natural history of PE, and reduced procedure time if compared with pharmacological thrombolysis. The clinical indications for percutaneous intervention in PE are discussed in the text. PMID- 12772131 TI - Applications of percutaneous mechanical thrombectomy in transjugular intrahepatic portosystemic shunt and portal vein thrombosis. AB - Portal vein thrombosis (PVT) is an uncommon cause for presinusoidal portal hypertension. PVT can be caused by one of three broad mechanisms: (1) spontaneous thrombosis when thrombosis develops in the absence of mechanical obstruction, usually in the presence of inherited or acquired hypercoagulable states; (2) intrinsic mechanical obstruction because of vascular injury and scarring or invasion by an intrahepatic or adjacent tumor; or (3) extrinsic constriction by adjacent tumor, lymphadenopathy or inflammatory process. Usually, several combined factors are necessary to result in PVT. The consequences of portal vein thrombosis are mostly related to the extension of the clot within the vein. Gastrointestinal bleeding from gastroesophageal varices is the most frequent presentation. Noninvasive imaging techniques are currently used for the screening of patients and the initial diagnosis of PVT. The invasive techniques are reserved for cases when noninvasive techniques are inconclusive, before percutaneous interventional treatment, or in preoperative assessment of patients who are candidates for surgery. Recanalization of the portal vein with anticoagulation alone may not be consistent or appropriate in highly symptomatic patients. Catheterization of the superior mesenteric artery (SMA) is helpful for diagnosis as well as for therapy by allowing the intra-arterial infusion of thrombolytic drugs in the same setting. Direct transhepatic portography allows precise determination of the degree of stenosis and extension within the portal vein, as well as pressure measurements. Thrombotic occlusions of the portal, mesenteric, and splenic veins can be managed by mechanical thrombectomy (MT) or pharmacologic thrombolysis. Underlying occlusions because of organized or refractory thrombus or fixed venous stenosis are best corrected by balloon angioplasty and stent placement. Access into the portal venous system can also be established through creating a transjugular intrahepatic portosystemic shunt (TIPS). Creating a TIPS is also important in the setting of PVT associated with cirrhosis to decompress portal hypertension and improve portal venous flow. PVT involving the portal, splenic, and/or mesenteric veins can also complicate a preexisting TIPS in which case the shunt can be readily used as therapy access. Several techniques may be used to recanalize the shunt and portal venous system, including thrombolytic therapy, balloon angioplasty/embolectomy, suction embolectomy, basket extraction of clots, and mechanical thrombectomy with a variety of devices. Advantages of MT include the potential to rapidly remove thrombus without the need for prolonged thrombolytic infusions, and reducing the potential life-threatening complications of thrombolytic therapy. Possible drawbacks include the risk of intimal or vascular trauma to the portal vein, which may promote recurrent thrombosis. PMID- 12772132 TI - Anticoagulation and neuraxial regional anesthesia: perspectives. PMID- 12772133 TI - Lipid reversal of bupivacaine toxicity: has the silver bullet been identified? PMID- 12772134 TI - Between innovation and proven value: achieving a balance in technical reporting. PMID- 12772135 TI - Regional anesthesia in the anticoagulated patient: defining the risks (the second ASRA Consensus Conference on Neuraxial Anesthesia and Anticoagulation). PMID- 12772136 TI - Lipid emulsion infusion rescues dogs from bupivacaine-induced cardiac toxicity. AB - BACKGROUND AND OBJECTIVES: We previously demonstrated in rats that intravenous infusion of a lipid emulsion increases survival in resuscitation from severe bupivacaine cardiac toxicity. The present studies were undertaken to determine if this method is similarly effective in a non-rodent model using a larger animal. METHODS: Bupivacaine, 10 mg/kg, was administered intravenously over 10 seconds to fasted dogs under isoflurane general anesthesia. Resuscitation included 10 minutes of internal cardiac massage followed with either saline or 20% lipid infusion, administered as a 4-mL/kg bolus followed by continuous infusion at 0.5 mL/kg/min for 10 minutes. Electrocardiogram (EKG), arterial blood pressure (BP), and myocardial pH (pHm) and pO2 (pmO2) were continuously measured. RESULTS: Survival after 10 minutes of unsuccessful cardiac massage was successful for all lipid-treated dogs (n = 6), but with no survivors in the saline controls (n = 6) (P <.01). Hemodynamics, PmO2, and pHm were improved during resuscitation with lipid compared with saline treatment in which dogs did not recover. CONCLUSIONS: We found that infusing a lipid emulsion during resuscitation from bupivacaine induced cardiac toxicity substantially improved hemodynamics, pmO2, and pHm and increased survival in dogs. PMID- 12772137 TI - Fascia iliaca compartment block for femoral bone fractures in prehospital care. AB - BACKGROUND AND OBJECTIVES: The fascia iliaca compartment block provides a faster and more consistent simultaneous blockade of the lateral cutaneous and femoral nerves than the "3 in 1" block. We studied the effectiveness of this technique for analgesia after a femoral bone fracture in pre-hospital care. METHODS: Patients with an isolated femoral shaft fracture were included. A fascia iliaca compartment block was performed on all of them. Twenty milliliters of lidocaine 1.5 % with epinephrine were injected under the fascia iliaca. The intensity of pain was measured using a simplified verbal scale (SVS) from 0 (no pain) to 4 (extreme pain). The SVS was noted before the block was performed, 10 minutes later, and then on admission to the trauma care center. Sensory blockade was evaluated using cold perception in the lateral, medial, and internal part of the thigh 10 minutes after block performance and on arrival at the trauma care center. RESULTS: Twenty-seven patients were enrolled in this study. The SVS was 3 (3-4) before the block, 1 (0-2) 10 minutes after the block, and 0 (0-1) when arriving at the trauma care center (P <.05). The SVS was lower when the internal part of the thigh was blocked. CONCLUSION: The fascia iliaca compartment block is a simple, inexpensive, and effective method of prehospital analgesia for femoral shaft fracture. A sensory block of the internal part of the thigh is an early predictive sign of optimal pain relief. PMID- 12772138 TI - Efficacy of 1% ropivacaine at sacral segments in lumbar epidural anesthesia. AB - BACKGROUND AND OBJECTIVES: It is suggested that the potency of 1% ropivacaine is comparable to that of 0.75% bupivacaine and higher than that of 2% lidocaine. Alkalinized lidocaine reportedly enhances the block of sacral segments during lumbar epidural anesthesia. We hypothesized that 1% ropivacaine might also block at the lumbosacral segments adequately during lumbar epidural anesthesia. METHODS: Forty-two patients undergoing lumbar epidural anesthesia at L4-5 or L5 S1 were randomly divided into 3 groups and received either 14 mL 2% lidocaine (lidocaine group), 2% lidocaine with epinephrine 1:200,000 and bicarbonate (lidocaine-epinephrine-bicarbonate group), or 1% ropivacaine (ropivacaine group). Pain threshold after repeated electrical stimulation was used to assess sensory block at the L2, S1, and S3 segments while motor block was evaluated using the modified Bromage Scale. RESULTS: Demographic data were comparable between the groups. Significant differences in the pH of each local anesthetic solution were found between the 3 groups. Pain thresholds at the S1 and S3 segments in the lidocaine-epinephrine-bicarbonate group were significantly higher and sensory block onset faster than in the other groups. However, no significant differences were found in either the pain threshold or the onset of sensory block of the L2 segment between the groups. No significant differences in the pain threshold, onset of sensory block, or Bromage Scale were found between the lidocaine and ropivacaine groups. CONCLUSIONS: We conclude that 1% ropivacaine does not improve block of sacral segments within 20 minutes following epidural ropivacaine administration. PMID- 12772139 TI - Free lidocaine concentrations during continuous epidural anesthesia in geriatric patients. AB - BACKGROUND AND OBJECTIVES: To evaluate the effects of aging on lidocaine pharmacokinetics, the plasma concentrations of total and free lidocaine and its metabolites were measured during continuous thoracic epidural anesthesia in middle-aged (age 41 +/- 9 years, n = 7) and elderly (age 72 +/- 2 years, n = 7) male patients. METHODS: After establishment of general anesthesia, 7 mL 1.5% lidocaine with epinephrine 1:200,000 was injected into the epidural space and subsequently infused at a rate of 5 mL/h for 5 hours. Plasma concentrations of total and free lidocaine, monoethylglycinexylidide (MEGX), and glycinexylidide (GX) were measured at 10, 15, 20, 30, 45, 60, 90, 120, 150, 180, 240, and 300 minutes after initial lidocaine injection using high-performance liquid chromatography (HPLC) with ultraviolet (UV) detection. RESULTS: The elderly group showed a stronger upward trend in the corrected free lidocaine concentrations and lower corrected total MEGX concentrations than the middle-aged group. CONCLUSIONS: Lidocaine metabolite activity in the elderly male patients was lower than that in the middle-aged male patients. Free lidocaine concentration is prone to increase in elderly patients. Caution must be exercised during continuous thoracic epidural anesthesia combined with general anesthesia in geriatric patients. PMID- 12772140 TI - A paramedian approach for epidural block: an anatomic and radiologic description. AB - BACKGROUND AND OBJECTIVES: Certain factors, such as intervertebral disc herniation, previous spinal surgery, and osteoarthritis may cause problems during the midline epidural procedure. The purpose of this study was to investigate the placement of the needle tip in an alternative paramedian approach in a cadaver, and radiographically measure the target interlaminar area at different levels (L3/L4; L4/L5; L5/S1) in different age groups. METHODS: The placement of epidural needles by an anesthetist using an alternative paramedian approach on 36 cadavers was evaluated by subsequent dissection along the needle to determine the location of the needle's terminal end. Radiographic measurements performed on anteroposterior lumbar spine x-rays in different age groups were used to determine the dimensions of the interlaminar area (horizontal, left vertical paramedian, and right vertical paramedian). RESULTS: Measurements of the interlaminar area significantly diminished with increasing age at L3/L4 (except for the horizontal measurement), L4/L5 (except for the vertical left paramedian measurement), as well as at L5/S1 in all measurements. CONCLUSION: This anatomically-based study confirms needle placement through the interlaminar space using an alternative paramedian approach and may explain the difficulties encountered during placement of an epidural needle with increasing age. PMID- 12772141 TI - Posterior perineal block with ropivacaine 0.75% for pain control during and after hemorrhoidectomy. AB - BACKGROUND AND OBJECTIVES: As perioperative pain management is a difficult challenge during hemorrhoidectomy, we tested the hypothesis that posterior perineal block (PPB) with local anesthetics alone is able to provide adequate pain control during and after surgery. METHODS: In a prospective, blinded, randomized study, we studied analgesic conditions and side effects of PPB in American Society of Anesthesiologists (ASA) I-II patients undergoing hemorrhoidectomy. Patients received general anesthesia (GA) either with PPB (0.75% ropivacaine, 40 mL (PPB group) or without PPB (control group). All patients received intravenous morphine patient-controlled analgesia (PCA) for postoperative pain control (morphine, 1.5 mg-boluses, 8-minute lockout interval). Intra- and postoperative opioids consumption was recorded, and pain assessments were performed at 1, 2, 4, 8, 12, and 24 hours using a visual analog scale (VAS). RESULTS: VAS scores were significantly lower during the first 8 postoperative hours in the PPB group as compared with the control group (P <.001). The PPB group required significantly less opioids during anesthesia (P <.001) and during the first postoperative day (P <.001) as compared with the control group. Time to first defecation and duration of hospitalization were identical in both groups. CONCLUSIONS: The present study shows that PPB with 40 mL 0.75% ropivacaine (300 mg) was a simple, effective, and safe method to provide better postoperative analgesia than PCA alone following surgical hemorrhoidectomy. In addition, PPB was shown to significantly reduce opioid consumption intraoperatively and during the first postoperative day. PMID- 12772142 TI - Adverse effects of local anesthetic infiltration on wound healing. PMID- 12772143 TI - Paravertebral approach to the brachial plexus: an anatomic improvement in technique. PMID- 12772144 TI - Complex regional pain syndrome type I after myocardial infarction treated with spinal cord stimulation. AB - OBJECTIVE: A rare case of Complex Regional Pain Syndrome (CRPS) type I after myocardial infarction (MI) and significant comorbid illness with few treatment options for pain control was successfully managed with the placement of a spinal cord stimulator (SCS). CASE REPORT: A 44-year-old man presented with left upper extremity burning pain after MI. His past medical history included insulin dependent diabetes mellitus, oxygen-dependent idiopathic pulmonary fibrosis, and recent coronary revascularization surgery. His pain was presumed to be related to his MI and a clinical diagnosis of CRPS type I (or reflex sympathetic dystrophy) was made. Facing limited medical and less invasive options for his pain relief, he underwent a spinal cord stimulation trial with excellent response. He had more than 70% pain relief from the spinal cord stimulation at the last follow-up, 2 years later. CONCLUSION: CRPS type I after MI can be difficult to treat because of other comorbid illnesses. SCS can be a safe and effective mode of therapy for patients facing limited treatment options. PMID- 12772145 TI - Cimetidine for chronic calcifying tendinitis of the shoulder. AB - BACKGROUND AND OBJECTIVES: Calcium deposits of the shoulder may persist for many years with resulting pain and impairment of mechanical function. The effects of different treatments vary significantly and do not show consistent and reliable long-term results. Cimetidine decreases calcium levels and improves symptoms in patients with hyperparathyroidism. We evaluated cimetidine as a treatment for chronic calcifying tendinitis of the shoulder in patients who did not respond to conservative treatment. METHODS: Cimetidine, 200 mg twice daily, was given orally for 3 months in 16 patients who did not respond to more than 6 months of conservative treatment. We recorded subjective, functional, and radiologic findings at 1 day before, at 2 weeks after, and at 2 and 3 months after the start of cimetidine. We also performed a follow-up study (4 to 24 months). RESULTS: After treatment, peak pain score (visual analogue scale: 0 - 100) decreased significantly from 63 +/- 13 to 14 +/- 19 (mean +/- SD, P <.01) and 10 patients (63%) became pain free. Physical impairment was also significantly improved. Calcium deposits disappeared in 9 patients (56%), decreased in 4 patients (25%), and did not change in 3 patients (19%). Follow-up data showed that improvement of symptoms was sustained. No recurrence or enlargement of calcium deposits was observed. Plasma concentrations of calcium and parathyroid hormone did not change significantly. CONCLUSIONS: Our results indicate that cimetidine is effective in treating chronic calcifying tendinitis of the shoulder; however, the mechanism by which cimetidine improves the symptoms is unknown. PMID- 12772146 TI - Lowest dose of spinal ropivacaine and adjuvant should be used in cesarean delivery. PMID- 12772150 TI - The charge-reverse analogy as an inspiration for the preparation of polydentate Lewis acidic boranes. PMID- 12772147 TI - Percutaneous electrode guidance and subcutaneous stimulating electrode guidance: modifications of the original technique. PMID- 12772151 TI - Triple bonds of the heavy main-group elements: acetylene and alkylidyne analogues of group 14. PMID- 12772152 TI - Watching photoinduced chemistry and molecular energy flow in solution in real time. AB - Energized molecules are the essential actors in chemical transformations in solution. As the rearrangement of bonds requires a movement of nuclei, vibrational energy is often the driving force for a reaction. Vibrational energy can be redistributed within the "hot" molecule, or relaxation can occur when molecules interact. Both processes govern the rates, pathways, and quantum yields of chemical transformations in solution. Unfortunately, energy transfer and the breaking, formation, and rearrangement of bonds take place on ultrafast timescales. This Review highlights experimental approaches for the direct, ultrafast measurement of photoinduced femtochemistry and energy flow in solution. In the first part of this Review, we summarize recent experiments on intra- and intermolecular energy transfer. The second part discusses photoinduced decomposition of large organic peroxides, which are used as initiators in free radical polymerization. The mechanisms and timescales of their decarboxylation determine the initial steps of polymerization and the microstructure of the polymer product. PMID- 12772153 TI - Analysis of interaction in patterned multienzyme layers by using scanning electrochemical microscopy. PMID- 12772154 TI - Towards "mechanochemistry": mechanically induced isomerizations of thiolate-gold clusters. PMID- 12772155 TI - Reduction of a 1-aza-1,3-diene to a 1-azabut-2-ene-1,4-diyl dianion: an unusual reaction course. PMID- 12772156 TI - Secondary isotope effects on the deslipping reaction of rotaxanes: high-precision measurement of steric size. PMID- 12772157 TI - cis-bromination of encapsulated alkenes. PMID- 12772158 TI - SmI2-induced umpolung of the C=N bond: first reductive conjugate addition of nitrones to alpha,beta-unsaturated esters. PMID- 12772159 TI - A conformational flexibility scale for amino acids in peptides. PMID- 12772160 TI - Optical logic operation based on polymer Langmuir-Blodgett-film assembly. PMID- 12772161 TI - Incorporation of selenomethionine into proteins through selenohomocysteine mediated ligation. PMID- 12772162 TI - Calix[n]bipyrroles: synthesis, characterization, and anion-binding studies. PMID- 12772163 TI - Stereoelectronic, strain, and medium effects on the protonation of cubylamine, a Janus-like base. PMID- 12772164 TI - From angstroms to micrometers: self-organized hierarchical structure within a polymer film. PMID- 12772165 TI - A self-assembled 2D molecule-based magnet: the honeycomb layered material [Co3Cl4(H2O)2[Co(Hbbiz)3]2]. PMID- 12772166 TI - A pseudorotaxane on gold: formation of self-assembled monolayers, reversible dethreading and rethreading of the ring, and ion-gating behavior. PMID- 12772167 TI - Remarkable positional discrimination in bistable light- and heat-switchable hydrogen-bonded molecular shuttles. PMID- 12772168 TI - Self-assembled receptors that stereoselectively recognize a saccharide. PMID- 12772169 TI - Living templates for the hierarchical assembly of gold nanoparticles. PMID- 12772170 TI - Direct-write dip-pen nanolithography of proteins on modified silicon oxide surfaces. PMID- 12772172 TI - The bitter truth: sensitivity to saccharin's bitterness predicts overactivity in highly arousable female dieters. AB - OBJECTIVE: The interaction between taste sensitivity and emotionality in rats provides a provocative view of hyperactivity. Rats that have been bred selectively for their reactivity to saccharin exhibit characteristic emotionality. When placed on restrictive diets, these rats exhibit excessive activity levels, relative to rats that are not sensitive to saccharin. Because humans who are highly arousable (i.e., reactive to environmental stimuli) also exhibit an increase in sensitivity to saccharin's bitterness, the current study evaluated whether women who are highly arousable, currently dieting, and sensitive to saccharin's bitterness engage in excessive exercise. METHOD: Participants completed a questionnaire packet, which assessed emotionality, eating patterns, and exercise patterns. On another occasion, they completed a body contour drawings handout, and their weight and height were measured. They also rated saccharin's bitterness and sweetness following a stressful event. RESULTS AND DISCUSSION: As hypothesized, sensitivity to saccharin's bitterness predicted overactivity in highly arousable female dieters, which reveals the multidimensionality of activity anorexia. PMID- 12772173 TI - A test of the continuity perspective across bulimic and binge eating pathology. AB - OBJECTIVE: This article examines the continuity/discontinuity perspective of eating pathology among 375 women seeking treatment. METHODS: Participants were categorized into five separate groups: obese nonbingers, subthreshold binge eating disorder (BED), BED, subthreshold bulimics, and bulimics. We tested whether differences in core eating pathology (drive for thinness, body dissatisfaction, current body image, body image ideal) and psychiatric symptoms (depression, interoceptive awareness) differentiated the groups quantitatively (supporting the continuity perspective) or qualitatively (supporting the discontinuity perspective). RESULTS: Our results, overall, supported the continuity perspective of eating pathology. A discriminant function analysis using the eating pathology and psychiatric symptom variables as predictor variables found that one primary factor differentiated the five groups on both core eating pathology and psychiatric variables. DISCUSSION: The implications of testing this model within a treatment-seeking sample are discussed. PMID- 12772174 TI - The relationship among binge severity, personality psychopathology, and body mass index. AB - OBJECTIVE: To determine the relationship among binge severity, personality psychopathology, and body mass index (BMI) among individuals with binge eating disorder (BED). METHOD: Participants were 50 individuals from the community who responded to advertisements for a medication treatment study of compulsive overeating and met BED criteria. Participants completed the Eating Disorder Examination and the Structured Clinical Interview for DSM-IV Personality Disorders. RESULTS: Binge eating severity was significantly and positively correlated with BMI and personality disorder symptomatology. The frequency of binge eating episodes, but not binge size, was associated with greater levels of personality psychopathology. BMI and personality psychopathology were unrelated. Twelve percent of the participants met full criteria for avoidant personality disorder (APD) and 10% met full criteria for obsessive-compulsive personality disorder (OCPD). DISCUSSION: Findings suggest that eating pathology, rather than weight, is related to personality psychopathology. APD and OCPD traits may be of particular relevance when considering treatment interventions for individuals with BED. PMID- 12772175 TI - Adverse effects of social pressure to be thin on young women: an experimental investigation of the effects of "fat talk". AB - OBJECTIVE: Experiments have found that pressure to be thin from the media promotes body dissatisfaction and negative affect, but the effects of social pressure to be thin have not been examined experimentally. Thus, this study tested whether social pressure to be thin fosters body dissatisfaction and negative affect. METHOD: Young women (N = 120) were randomly assigned to a condition wherein an ultra-thin confederate complained about how fat she felt and voiced intentions to lose weight or a control condition wherein she discussed a neutral topic. RESULTS: Exposure to social pressure to be thin resulted in increased body dissatisfaction but not negative affect. The effects were not moderated by initial thin-ideal internalization, body dissatisfaction, or social support. DISCUSSION: Results support the assertion that peer pressure to be thin promotes body dissatisfaction but suggest that this factor may not contribute to negative affect. PMID- 12772176 TI - Anger and core beliefs in the eating disorders. AB - OBJECTIVE: The link between emotion and eating pathology has long been established, but relatively little is known about the role of anger, partly because the existing literature has tended to concentrate on anger as a unitary construct. Nor is there any understanding of the cognitive factors that drive this affect in the eating disorders. This study had two aims: to determine levels of different facets of anger across eating disorder diagnoses and behaviors; and to investigate whether facets of anger are related to the individual's negative core beliefs. METHOD: The sample consisted of 140 women who met DSM-IV criteria for eating disorders, and 50 female control participants (university undergraduates). The women completed self-report questionnaires of anger levels and unhealthy core beliefs, and the presence of bulimic behaviors was recorded at assessment. RESULTS: The eating-disordered women had higher levels of state anger and anger suppression, particularly if the diagnosis included bulimic symptoms. Different aspects of anger were associated with specific bulimic behaviors. Unhealthy core beliefs were associated with higher levels of trait anger in both groups but with anger suppression in the clinical women only. DISCUSSION: Suggestions are made regarding ways in which state anger and anger suppression might be understood and treated in women with eating disorders. PMID- 12772177 TI - Gender differences in implicit weight identity. AB - OBJECTIVE: This study examined gender differences in explicit and implicit attitudes toward overweight and explicit and implicit weight identity. METHOD: Normal weight women (n=22) and men (n=20) and overweight women (n=20) and men (n=21) completed the Implicit Association Test and portions of the Eating Disorders Questionnaire. RESULTS: Although explicit and implicit anti-fat attitudes were ubiquitous, gender differences emerged for weight identity. Both men and women provided accurate explicit appraisals of their weight status. However, men implicitly identified themselves as light regardless of actual weight status. Women's implicit weight identity was associated with their actual weight status, explicit weight appraisal, and implicit self-esteem. DISCUSSION: These findings may provide additional insight into why men are underrepresented among those seeking weight loss and why women are at increased risk for developing eating disorders. PMID- 12772179 TI - Body image and eating disturbance among south Asian-American women: the role of racial teasing. AB - OBJECTIVE: Acculturation and loss of ethnic identification have been proposed as risk factors for eating and body image disturbance among women of color. This study investigated whether being teased about racial or ethnic features might also play a role in these disturbances in minority women. METHODS: One hundred twenty-two college women of South Asian descent completed questionnaire measures of disturbed eating behavior, body image dissatisfaction, distress, self-esteem, acculturation, ethnic identification, and racial teasing. RESULTS: History of hurtful racial teasing, but not acculturation or ethnic disidentification, was associated with disturbed eating and body image, even after controlling for distress, self-esteem, and body mass. CONCLUSION: The psychological impact of racial teasing may be a potent but neglected source of eating and body image disturbance among minority women. PMID- 12772178 TI - A comparison of cognitive and behavioral symptoms between Mexican and American eating disorder patients. AB - OBJECTIVE: Mexican and U.S. patients with anorexia nervosa or bulimia (complete and partial syndromes) were compared on severity and types of preoccupations and rituals related to eating disorders and the motivation to change. METHOD: One hundred seventy-four patients who met DSM-IV criteria for anorexia nervosa, bulimia nervosa, or ED-NOS participated. Eighty-seven subjects entered treatment at the Instituto Nacional de Psiquiatria "Ramon de la Fuente" in Mexico City and were matched with 87 patients treated at the New York Presbyterian Hospital. Patients were interviewed with the Yale-Brown-Cornell Eating Disorder Scale (YBC EDS) (English or Spanish version). RESULTS: All YBC-EDS scores were higher for the Mexican group, which also had a greater number of current preoccupations and rituals. U.S. and Mexican patients were quite similar in their endorsement of current preoccupations, but the Mexican group was more likely to have rituals in all checklist categories, and the rituals were more egosyntonic. More U.S. patients had received previous treatment for their eating disorder. A negative correlation was obtained between the amount of previous treatment and motivation to change the preoccupations and rituals (higher scores indicating lower motivation). DISCUSSION: Cultural factors may influence characteristics of eating disorders. The previous treatment experiences of the U.S. patients seemed to positively influence their motivation for further treatment. PMID- 12772180 TI - History of anorexia nervosa in bulimic patients: its influence on body composition. AB - OBJECTIVE: To analyze the body composition of a group of patients fulfilling DSM IV criteria for bulimia nervosa (BN), comparing the patients who had a history of DSM-IV anorexia nervosa (AN) with those without such a history to determine (1) whether both groups differed in terms of body composition and (2) whether the differences between groups could be the consequence of the past condition. METHODS: The sample consisted of 104 patients, 43 with prior AN (BN-AN group) and 61 without a history of AN (BN-nonAN). The patients were assessed using anthropometric and bioelectrical methods: height and weight, body circumferences, abdominal diameter, skinfold thickness, and body impedance analysis. In a later phase and to test the influence of current low weight on the results, two experimental groups were created: the first group was composed of the patients from the BN-AN group who had a normal body mass index (BMI range in this group, 20.1-23.4 kg/m2; n = 23); the second group consisted of 23 patients from the BN nonAN group in the same BMI range. In both phases unpaired sample t tests were performed for statistical analysis. RESULTS: More than 40% of the bulimic patients with a history of AN had a BMI less than 20. They had a lower percentage of body fat, lower muscle mass, and higher percentage of extracellular water. Nevertheless, most of these differences disappeared in the second phase of the analysis, when only the patients within a normal weight range were compared. DISCUSSION: According to these results, a significant number of bulimic patients with a history of AN tend to retain some clinical traits of the past condition and could be viewed as remaining in a "subclinical anorexic status": they are thinner and seem to have less difficulties in maintaining low weight than patients without a history of AN. Nevertheless, in patients who have reached a normal weight after AN, all these differences disappear. This fact raises some important questions related to the boundaries between AN and BN, the shifting from one to another, or when AN patients really recover. PMID- 12772181 TI - Resting energy expenditure and plasma leptin levels in adolescent girls with anorexia nervosa. AB - OBJECTIVE: Our aim was to examine the relationship between resting energy expenditure (REE) and circulating leptin levels during long-term refeeding in adolescent girls with anorexia nervosa (AN). METHOD: We measured REE, the plasma leptin level, the free T(3) level, body surface area (BSA), and dietary energy intake in AN adolescents before and 8 weeks after refeeding therapy. RESULTS: All parameters in AN subjects before the refeeding therapy were significantly lower than those in AN subjects after the therapy and in the controls. There were no significant differences in REE and BSA between posttherapy AN subjects and controls, but plasma leptin levels in the posttherapy AN subjects were still significantly lower than those in the controls. Significant correlations between REE and BSA were detected in the AN subjects both before and after the refeeding therapy, but REE did not correlate significantly with plasma leptin level in either of the AN groups. DISCUSSION: Our results suggest that the REE in AN subjects after long-term refeeding therapy is not associated with the changes in plasma leptin levels but with BSA. PMID- 12772182 TI - Body size estimation: testing a new mirror-based assessment method. AB - OBJECTIVE: The aim of the current study was to compare a new ecologically valid method of body size estimation with an existing, commonly used method. METHOD: The new method uses a "mirror-sized" image and measures perception rather than memory of body size. Fifty-five healthy women participated in the study. RESULTS: Body size estimations with the new method were significantly larger than with the existing method (p <.001). This difference was specific to body size and was not seen for estimation of neutral images. DISCUSSION: The results of the study show that different body size estimations are made with the new more ecologically valid method compared with the existing method. This finding has important implications for research into the factors influencing the perception of body size. PMID- 12772183 TI - Sociotropy and bulimic symptoms in clinical and nonclinical samples. AB - OBJECTIVE: The purpose of this study is to examine the relation between sociotropy and bulimic symptoms. Studies of interpersonal functioning among individuals with bulimia nervosa consistently reveal issues of social dependency, need for approval, and fear of rejection. These themes are conceptually related to sociotropy, a cognitive-personality factor that has been implicated in the development and maintenance of depression. Individuals high in sociotropy are keenly invested in attaining others' approval and avoiding social rejection. METHODS: The relationship between sociotropy and bulimic symptoms was examined in two samples of women: undergraduate women and community women seeking treatment at a private eating disorder facility. RESULTS: In both samples, sociotropy was significantly associated with bulimic symptoms beyond the shared relation with depressed mood. DISCUSSION: Findings are discussed in terms of the maintenance and treatment of bulimia nervosa. PMID- 12772184 TI - The effects of restraint, gender, and body mass index on the accuracy of self reported weight. AB - OBJECTIVE: Previous research has shown that women, restrained eaters, and overweight/obese individuals have a tendency to underreport their weight. This study investigated gender, dietary restraint, and BMI to determine which is the best predictor of errors in self-reported weight. METHOD: A total of 241 college students completed questionnaires pertaining to height and weight and then were weighed in the laboratory. RESULTS: Although overall students were fairly accurate in reporting their weight, restraint status was the best predictor of weight underestimation. DISCUSSION: These results suggest that restrained eaters, regardless of gender or BMI, have a tendency to underestimate their weight. PMID- 12772185 TI - Prognostic value of circulating prostate cells in patients with a rising PSA after radical prostatectomy. AB - BACKGROUND: To predict poor outcome in patients with a biochemical recurrence (rising PSA) after radical prostatectomy (RP), urologists rely primarily on Gleason score, PSA doubling time, and time from surgery to biochemical (i.e., PSA) recurrence. In the present study, we assess the value of RT-PCR detection circulating prostate cells in blood of patients with a rising PSA. METHODS: RNA from blood samples was obtained from 55 patients with a rising PSA and from 45 patients without evidence of biochemical failure (PSA < 0.1 ng/ml). Both groups were matched for age, Gleason score, pT stage, and interval between radical prostatectomy and PCR testing. RESULTS: PSA positive cells were detected in 1/45 (2%) patients without a PSA recurrence and 19/55 (34%) patients with a PSA recurrence. In the rising PSA group, mean PSA doubling time was significantly shorter in patients with positive RT-PCR (5 months) than in patients with negative RT-PCR (16 months; P = 0.001). An earlier onset of recurrence was also detected in patients with a positive RT-PCR (31 months for positive RT-PCR vs. 50 months for negative RT-PCR) but this result did not achieve statistical significance (P = 0.102). Salvage radiation therapy was administered in 15 patients. Three of the five patients with a positive RT-PCR progressed during radiotherapy whereas 7 of the 10 patients with a negative RT-PCR obtained a complete response and none have progressed. CONCLUSIONS: These preliminary results suggest that RT-PCR detection of prostate cells in blood of patients after RP correlates with rapidly progressing biochemical failure after RP. PMID- 12772187 TI - Response of patients with advanced prostatic cancer to administration of somatostatin analog RC-160 (vapreotide) at the time of relapse. AB - BACKGROUND: The aim of this study was to evaluate the effects of administration of the somatostatin analog RC-160 (vapreotide) at the time of relapse in patients with androgen independent prostate cancer. METHODS: Our study included 13 patients with biopsy-proven prostate cancer, stage D3. Eight patients had been treated with a depot formulation of the agonist D-Trp-6-LH-RH, with a median remission time of 68 (range 48-102 months). Five patients were initially treated by surgical orchiectomy, but relapsed after a median time of 33 months (range 17 91 months). A new remission period with a median duration of 10 months (range 2 29 months) was induced with Ketoconazole in the orchiectomy group. At the relapse time, all the patients received 1 mg of vapreotide t.i.d., by subcutaneous route, in addition to D-Trp-6-LH-RH, or Ketoconazole in the orchiectomy group. RESULTS: Eight of 13 patients demonstrated clinical improvement after 3 months of therapy with vapreotide, six showing a decrease in serum prostate specific antigen (PSA) from 234.5 +/- 308.5 to 68.2 +/- 60.5 ng/ml (mean decline 71 +/- 8%; P < 0.05). Two additional patients presented a fall in serum prostatic acid phosphatase (PAP). Responding patients showed a decrease in the bone pain score from 2.62 +/- 0.48 to 0.37 +/- 0.69 and an increase in the Karnofsky performance status from 72.3 +/- 4.21 to 83.6 +/- 23.2 (P < 0.05). In accord with the ECOG criteria, two patients had a complete response; four had partial response, and two had a stable response. Four patients did not respond and one was not evaluable. Two patients died in remission, one at 16 months due to myocardial infarction and the other at 24 months due to pneumonia. Three patients relapsed at 5, 17, and 19 months respectively. Three patients who have been followed-up for more than 3 years continued in remission (79, 45, and 45 months) respectively. Vapreotide was well tolerated, only three patients having transitory mild diarrhea. CONCLUSIONS: Our results indicate that therapy with the somatostatin analog vapreotide at the time of relapse can induce objective clinical responses in some patients with prostate cancer who are refractory to androgen ablation induced by LH-RH analogs or orchiectomy. PMID- 12772186 TI - Expression and function of pro-inflammatory interleukin IL-17 and IL-17 receptor in normal, benign hyperplastic, and malignant prostate. AB - INTRODUCTION AND OBJECTIVES: To investigate factors involved in inflammation of the prostate besides IL-15, we screened prostatic cells and tissues for IL-17 and IL-17 receptor expression. METHODS: Normal prostate (n = 1), BPH (n = 19), and carcinoma (CaP, n = 12) specimens were screened for IL-17, IL-17 receptor, CD45, IL-6, and IL-8 mRNA expression. The carcinoma cell lines DU145, PC3, LNCaP, and BPH-epithelial (EC), stromal cell (SC) preparations, and BPH-T-cell lines were analyzed for IL-17 production by RT-PCR and ELISA. The effect of IL-17 on IL-6, IL-8, TGF-beta1, and fibroblast growth factor (FGF-2) mRNA expression and/or release of SC was analyzed using real-time PCR and/or ELISA. Immunohistochemistry was used to localize both IL-17 and IL-17 receptor. RESULTS: In the normal prostate, IL-17 expression was very weak and restricted to lymphocytes. In 79% of BPH and 58% of CaP specimens, IL-17 mRNA and protein expression was increased. IL 17 mRNA expression could be shown for activated BPH-T-cells and to some extend for BPH-EC. Expression of IL-17 receptor was ubiquitous. Release of IL-17 was shown only for activated BPH-T-cells. IL-17 stimulated expression of IL-6 (13 fold) and IL-8 (26-fold) by prostatic BPH-SC. In situ, however, the amount of IL 17mRNA in BPH-tissue did not correlate with the amount of IL-6 and IL-8 mRNA. In CaP tissue, significant correlation was found only between the amount of IL-6 and IL-8 mRNA. CONCLUSIONS: Activated BPH-T-cells abundantly express IL-17. The increase of IL-17 in BPH-tissues goes hand in hand with elevated levels of IL-15, a pro-inflammatory cytokine with T-cell growth factor properties. A clinical relevance of increased IL-17 expression under pathological conditions is suggested by the demonstration of significant upregulation of IL-6 and IL-8 production of prostatic SC by IL-17. PMID- 12772188 TI - Expression of androgen receptor coactivators in normal and cancer prostate tissues and cultured cell lines. AB - BACKGROUND: In prostate cancer cell lines, androgen receptor (AR) coactivators modulate the transcriptional activity of AR. However, very little is known about their expression in normal prostate tissue and during progression to cancer. METHODS: AR and coactivators ARA54, ARA55, ARA70, and SRC1 RNA were analyzed by RT-PCR in normal and tumoral tissues of the same prostate, in prostate cell lines, and after hormonal treatments of prostate epithelial cells. RESULTS: AR coactivators were expressed in normal and tumoral tissues and in cultured prostate cells; only ARA55 expression was decreased in tumoral relative to normal tissue of all seven prostates analyzed. It was not expressed in LNCaP and DU145 cancer cells and low in PNT2 immortalized cells in which all coactivator's expression were down regulated by DHT and up regulated by E2. In addition, coactivator's expression was increased in hyperplastic relative to normal prostate fibroblasts. CONCLUSIONS: ARA55 is both an AR coactivator and a focal adhesion protein (Hic-5). Its role in the progression of prostate carcinoma may therefore involve these two different functions. Its decrease in cancer tissue suggests that it plays a different role than that expected, namely, facilitate cell proliferation and therefore mobility and metastasis. PMID- 12772189 TI - A low-fat diet and/or strenuous exercise alters the IGF axis in vivo and reduces prostate tumor cell growth in vitro. AB - BACKGROUND: Prostate cancer is the most common solid-tumor cancer in US males but is rare in Asian males. When Asian men adopt the US lifestyle, clinical prostate cancer increases greatly. Epidemiological data from men in the US indicate that regular activity may reduce the risk for prostate cancer. METHODS: Serum was obtained from three groups of similar-aged men, Control, Diet and Exercise, and Exercise alone were used to stimulate LNCaP cells in culture. Growth and apoptosis of tumor cells were measured. Serum samples were also used to measure insulin, IGF-1, IGFBP-1. RESULTS: The Diet and Exercise and the Exercise alone groups had lower serum insulin and IGF-1 but higher IGFBP-1 compared to Controls. LNCaP cell growth was reduced in both groups compared to Control and there was a major increase in apoptosis of tumor cells. CONCLUSIONS: A low-fat diet and/or intensive exercise results in change in serum hormones and growth factors in vivo that can reduce growth and induce apoptosis of LNCaP prostate tumor cells in vitro. PMID- 12772190 TI - Recent trends in mortality from benign prostatic hyperplasia. AB - BACKGROUND: We have considered trends in mortality from benign prostatic hyperplasia (BPH) over the last decades in Europe and, for comparative purposes, the USA and Japan. METHODS: Cancer death certification data for benign prostatic hyperplasia were derived from the World Health Organisation database. RESULTS: Between the early 1950s and the late 1990s, overall mortality from BPH in the European Union (EU) fell from 5.9 to 3.5 per million, and the decline since the late 1950s was over 96%. Comparable falls were observed in the USA and Japan, and BPH mortality rates in the late 1990s were lower than in the EU (1.8/10(6) in the USA, 1.4 in Japan). BPH mortality trends were downwards also in the Eastern Europe, although rates in the late 1990s were about fourfold higher than in the EU. CONCLUSION: BPH rates have been steadily declining in developed countries. The excess BPH mortality in Eastern Europe indicates the scope for further reduction too. PMID- 12772191 TI - Recombinant prostate specific antigen inhibits angiogenesis in vitro and in vivo. AB - BACKGROUND: Prostate specific antigen (PSA) is a kallikrein family member with serine protease activity commonly used as a diagnostic marker for prostate cancer. We recently described anti-angiogenic properties of PSA [Fortier et al.: JNCI 91:1635-1640]. METHODS: Two forms of PSA were cloned and expressed in Pichia pastoris: one, an intact PSA with an N-terminus of IVGGVS em leader; the second, an N-1 PSA variant. The recombinant proteins were tested for serine protease activity and for anti-angiogenic activity in vitro and in vivo. RESULTS: The rate of substrate hydrolysis by the intact recombinant PSA was similar to that of PSA isolated and purified from human seminal plasma. In contrast, the N-1 PSA variant lacked serine protease activity. In an endothelial cell migration assay, the concentration that resulted in 50% inhibition (IC(50)) was: 0.5 microM for native PSA, 0.5 microM for intact recombinant protein, and 0.1 microM for the N-1 variant PSA. Both the intact recombinant and the N-1 recombinant PSA inhibited angiogenesis in vivo. CONCLUSIONS: Purified recombinant PSA inhibits angiogenesis, proving the concept that PSA is an anti-angiogenic, and serine protease activity, as determined by synthetic substrate hydrolysis, is distinct from the anti-angiogenic properties of PSA. PMID- 12772192 TI - Prostatic androgen repressed message-1 (PARM-1) may play a role in prostatic cell immortalisation. AB - BACKGROUND: Prostatic androgen-repressed message-1 (PARM-1) has been cloned from the prostate. The transcript of the PARM-1 gene is overexpressed during regression of the prostate after androgen withdrawal. The regulation of PARM-1 by androgens is limited to this organ. We have studied the effects of PARM-1 overexpression in malignant prostate cells. METHODS: The PARM-1 cDNA was introduced into the rat cancer cell line MAT LyLu along with a doxycycline dependent regulator. RESULTS: Maximal expression of PARM-1 (fivefold induction) was achieved by incubating the cells with 2 microM doxycycline for 48 hr. A study investigating the effect of PARM-1 overexpression on the transcription of 588 genes has shown that the TLP1 gene (encoding rat telomerase protein component 1) was the most up-regulated (fourfold). In addition, a dose-dependent increase in telomerase activity was observed in cells overexpressing PARM-1. In vivo, the androgen-deprived prostate showed an increased TLP1 level and increased telomerase activity. CONCLUSIONS: Increased telomerase activity is often associated with the immortalisation of cancer cell lines, particularly prostatic ones. This could mean that PARM-1 is involved, via increased telomerase activity, in a survival program enabling certain prostatic cells to resist apoptosis, thus conferring a selective advantage to pre-cancerous or cancerous cells. PMID- 12772193 TI - Establishment of a prostatic small-cell carcinoma cell line (SO-MI). AB - BACKGROUND: Prostatic small-cell carcinoma is an extremely rare, highly aggressive disease. We established a cell line from this tumor. MATERIALS AND METHODS: Tumor tissue obtained from a 24-year-old Japanese man was used to establish the cell line. Cultured cells and tumors transplanted into nude mice were characterized by histologic, immunohistologic, immunocytologic, and molecular biologic methods. RESULTS: An immortal culture cell line (SO-MI) was successfully established. SO-MI cells adhered weakly to plastic surfaces in vitro, showing a 52- to 72-hr doubling time. SO-MI cells were heterotopically and orthotopically transplantable in nude mice. The cells were immunoreactive for NSE, chromogranin A, and NCAM, but not for ACTH, calcitonin, serotonin, gastrin, insulin, glucagons, LCA, EMA, PAP, PSA, androgen receptor, and p53. SO-MI cells secreted NSE in vitro and in vivo. SO-MI cells at passage 30 contained 50-59 chromosomes with a modal number of 55. PCR suggested that the p53 gene was deleted in SO-MI cells. RT-PCR detected no mRNA encoding androgen receptor in these cells. CONCLUSIONS: SO-MI cells retain the neuroendocrine nature of the original tumor, and should be useful in studying possible etiologies and new treatments. PMID- 12772194 TI - Breast cancer-have we been chasing the wrong target? PMID- 12772196 TI - Emerging patterns of practice in the implementation and application of sentinel lymph node biopsy in breast cancer patients in Canada. PMID- 12772197 TI - Emerging patterns of practice in the implementation and application of sentinel lymph node biopsy in breast cancer patients in the United States. PMID- 12772198 TI - Surgeon volume compared to hospital volume as a predictor of outcome following primary colon cancer resection. AB - BACKGROUND AND OBJECTIVES: A strong association between high hospital procedure volume and survival following colon cancer resection has been demonstrated. However, the importance of surgeon case volume as a determinant of outcome has been less well studied, and it is unclear whether hospital or surgeon volume is the more powerful predictor of outcomes. METHODS: A retrospective population based cohort study utilizing the Surveillance, Epidemiology, and End Results (SEER)-Medicare linked database identified 24,166 colon cancer patients aged 65 years and older who had surgery for a primary tumor diagnosed in 1991-1996 in a SEER area. Hospital and surgeon-specific procedure volume was ascertained based on the number of claims submitted over the 6-year study period. Outcome measures were mortality at 30 days and 2 years, overall survival, and the frequency of operations requiring an intestinal stoma. Age, sex, race, comorbid illness, cancer stage, socioeconomic status, emergent hospitalization, and the presence of obstruction/perforation were used to adjust for differences in case-mix. RESULTS: After adjusting for surgeon procedure volume, high hospital procedure volume remained a strong predictor of low post-operative mortality rates (P < 0.001 for each outcome with and without adjustment for surgeon procedure volume). Surgeon specific procedure volume was also an important predictor of surgical outcomes (P = 0.002 for 30-day mortality, P = 0.001 for 2-year mortality), although this effect was attenuated after adjusting for hospital volume (P = 0.03 for 30-day mortality, P = 0.02 for 2-year mortality). Hospital volume and surgeon volume were each an important predictor of the ostomy rate. Among high volume institutions and surgeons, individual providers with unusually high ostomy rates could be identified. CONCLUSIONS: Both hospital and surgeon-specific procedure volume predict outcomes following colon cancer resection; but hospital volume may exert a stronger effect. Therefore, efforts to optimize the quality of colon cancer surgery should focus on multidisciplinary aspects of hospital care rather than solely on intraoperative technique. PMID- 12772200 TI - The role of preoperative ultrasound scan in detecting lymph node metastasis before sentinel node biopsy in melanoma patients. AB - BACKGROUND AND OBJECTIVES: To evaluate the efficacy of preoperative ultrasound (US) scanning in identifying lymph node metastasis before sentinel node biopsy (SNB), we conducted a prospective study on 125 patients with primary cutaneous melanoma (CM). METHODS: We prospectively enrolled 125 patients with >1 mm thick CM and candidate for SNB. Preoperatively, patients underwent US scanning of regional lymphatic basins and FNA of suspected lymph nodes (LN). All patients underwent lymphatic mapping and SNB. RESULTS: Combined with fine-needle aspirate (FNA) of suspect LN, US scan allowed the correct preoperative detection of 12 out of 31 histologically positive lymphatic basins, specificity and sensitivity being 100 and 39%, respectively. The false negative rate (61%) was mainly linked to tumor deposits less than 2 mm in diameter, which can be considered the current spatial resolution limit of this technique. CONCLUSIONS: Preoperative US scan could reduce the number of SNB, thus avoiding the stress of this surgical procedure in approximately 10% of patients and reducing health care costs. As a non-invasive and relatively inexpensive technique, lymph node US scan can be part of the preoperative staging process of patients' candidate for SNB in order to avoid unnecessary surgical procedures. PMID- 12772201 TI - Heterogeneic expression of estrogen receptor between the primary tumor and the corresponding involved lymph nodes in patients with node-positive breast cancer and its implications in patient outcome. AB - BACKGROUND AND OBJECTIVES: The estrogen-receptor (ER) status of breast cancers has typically been evaluated in primary tumors (PTs), and the influence of ER expression in the involved lymph nodes (LNs) on prognosis of the patients with node-positive breast cancer remains unclear. The expression of ER was compared between PT and corresponding involved LNs in patients with node-positive breast cancer. METHODS: Eighty-seven patients with node-positive breast cancer were immunohistochemically assessed to measure the expression of ER in PT and involved LN with anti-ER monoclonal antibody. RESULTS: Thirty-five (40.2%) of PTs and 26 (29.9%) of involved LNs were evaluated as ER(+). The ER expression in PTs was significantly correlated with that in involved LNs (P < 0.0001), and the ER expression was almost the same in both the PT and corresponding involved LN in 66 cases (75.9%): both (+), 20 (23.0%); and both (-), 46 (52.9%). However, ER expression differed in 21 cases (24.1%) between involved LN and the corresponding PT: PT-ER(+)/LN-ER(-), 15 (17.2%); and PT-ER(-)/LN-ER(+), 6 (6.9%). The survival of patients with ER(+)-PT was significantly better than that of ER(-)-PT patients (P = 0.0086), but ER expression in involved LN did not have any significant effect on patient survival. Furthermore, when survival periods were grouped by coexpression of ER in the PT and involved LN, the best survival was seen in the PT-ER(+)/LN-ER(-) group (P < 0.001 vs. others). In the patients, who received adjuvant endocrine therapy, the survival of the PT-ER(+) group was significantly better than that of PT-ER(-) group (P = 0.0145), but LN-ER expression did not show any significant influence on the survival rate. Multivariate analysis demonstrated that clinical stage and ER-expression in the PT were significant variables, but ER expression in the involved LN was not. CONCLUSIONS: The ER expression was discordant between PT and involved LN in about 24% of the breast cancers evaluated, and ER expression in involved LNs had less influence on patient prognosis than ER expression in the corresponding PT. PMID- 12772202 TI - Clinicopathological characteristics of superficial spreading type early gastric cancer. AB - BACKGROUND AND OBJECTIVES: Superficial spreading type early gastric cancer is characterized by its atypical growth pattern and occasionally indistinct tumor margin. Because it is a rare form of early gastric cancer, the clinicopathological details are not apparent. The aim of this study was to clarify the clinicopathological features of the superficial spreading type of early gastric cancer. METHODS: A retrospective study was conducted in 1,062 surgically resected patients with early gastric cancer. Hospital records were compared between patients with superficial spreading type early gastric cancer and those with more common types of early gastric cancer. RESULTS: Sixty-nine patients (6.9%) had superficial spreading lesions. The male to female ratio was 1.2:1. The most frequent histological type was signet-ring cell carcinoma (32%). The distinguishing histopathological features were submucosal invasion (67%), lymphatic invasion (32%), and lymph node metastasis (30%). There were discrepancies in tumor area between surgical findings and pathological diagnosis in 24 patients (35%) with superficial spreading type. More extensive lymph node dissection was performed and all patients survived in the group with superficial spreading lesions. CONCLUSIONS: The most appropriate treatment for the superficial spreading type of early gastric cancer is wide surgical resection with extensive lymph node dissection. PMID- 12772203 TI - Apolipoprotein D expression in cutaneous malignant melanoma. AB - BACKGROUND AND OBJECTIVES: Apolipoprotein D (Apo D) is a protein component of the human plasma lipid transport system, and an androgen-regulated protein in both breast and prostate cancer cell lines. Our goal was to evaluate the expression of Apo D in malignant cutaneous melanomas, as well as to assess its possible relationship to clinical and pathological parameters. METHODS: Apo D expression was analyzed in 32 paraffin-embedded tissues from patients with invasive cutaneous malignant melanomas, in 8 samples from in situ melanoma, and in 10 samples from 10 benign lesions (4 dermal melanocytic nevi, 4 compound melanocytic nevi, and 2 dysplastic melanocytic nevi), using immunohistochemical techniques. RESULTS: The benign lesions were consistently negative for Apo D, whereas 3 of the 8 "in situ" melanomas (37.5%) and 12 of the 32 invasive melanomas (37.5%) showed positive immunostaining for Apo D. The percentage of Apo D-positive tumors was significantly higher in nodular than in superficial spreading melanomas (P = 0.011) and in melanomas with vertical growth phase than in melanomas with radial growth phase (P = 0.02). In addition, the percentage of Apo D-positive tumors was positively and significantly correlated with Clark's level of invasion (P = 0.046). CONCLUSIONS: Apo D may be a new prognostic factor of unfavorable evolution in cutaneous malignant melanoma. PMID- 12772204 TI - Clinicopathologic and immunohistochemical study of surgically treated primary gastric MALT lymphoma. AB - BACKGROUND AND OBJECTIVES: B-cell MALT lymphoma is a well-recognized entity and its characterization as low-grade (LG) and high-grade (HG) lymphoma has been widely accepted. In the present study we reviewed a series of 95 surgical specimens of primary gastric MALT lymphoma selected between 1979 and 1998. Immunohistochemical expression of p53, bcl-2, and Ki67 and Helicobacter pylori (Hp) infection was evaluated, along with a correlation with clinical outcome. METHODS: A morphologic and immunohistochemical analysis, including p53, bcl-2, and Ki67 expression, was carried out in all cases. A complete follow-up was obtained in 49 patients and in these cases a survival analysis was performed. RESULTS: bcl-2 protein was highly expressed in 25 of 25 assessed LG tumors and in 20 of 24 assessed HG tumors. p53 protein was expressed in 13 of 25 assessed LG tumors and in 21 of 24 assessed HG tumors. High proliferation rate as expressed by Ki67 was detected in 15 of 25 assessed LG tumors and in 23 of 24 assessed HG tumors. Hp infection was detected in 11 of 16 assessed LG tumors and 2 of 10 assessed HG tumors. Median survival rates were 72 months for LG tumors and 24 months for HG tumors. CONCLUSIONS: A significant inverse relationship between Hp infection and histological grade was found. High p53 expression and high proliferation rate correlated with HG tumors. However, a correlation between p53, bcl-2, and Ki67 expression with clinical outcome was not found. PMID- 12772205 TI - Soluble Fas (sFas) and soluble Fas ligand (sFas-L) balance in laryngeal carcinoma before and after surgical treatment. AB - BACKGROUND AND OBJECTIVES: Fas and its specific ligand (Fas-L), both of which are involved in apoptosis, exist in membrane-bound and soluble forms. The soluble forms (sFas and sFas-L) have been observed in various tumours, but their clinical significance has not yet been clarified. The aim of this study was to assess serum sFas and sFas-L levels in patients with laryngeal squamous cell carcinoma (LSCC) and their possible correlations with surgical treatment. METHODS: Serum sFas and sFas-L levels were determined by ELISA in samples taken from 26 LSCC patients on the day before surgery (T0), and 2 weeks (T1) and 6 months after surgery (T2), and in samples taken from 35 healthy volunteers. RESULTS: The mean serum sFas levels in the 35 healthy volunteers and the 26 LSCC patients at T0 were respectively 5941 +/- 411 pg/ml and 6290 +/- 652 pg/ml (P = 0.63), and the mean serum sFas-L levels were 0.1 +/- 0.05 ng/ml and 2.95 +/- 0.8 ng/ml (P < 0.0001). After surgery, there was a statistically significant decrease in sFas at both T1 (P < 0.05) and T2 (P < 0.01), and in sFas-L at T2 (P < 0.01). CONCLUSIONS: The decrease in sFas and sFas-L levels after surgery suggest that they may be produced by or closely linked to tumour cells. Larger prospective clinical studies of patients with LSCC will be needed to establish the clinical significance of sFas and sFas-L, as reported for other neoplasms. PMID- 12772207 TI - Superelastic Ti-Ni alloy wire intramedullary nails for metastatic femoral pathologic fracture: a case report. PMID- 12772206 TI - Current management of mucosal melanoma of the head and neck. AB - While mucosal-based melanomas of the head and neck region are uncommon lesions, when they do arise they usually follow an inexorably aggressive course. Experience with these tumors is, necessarily, limited; as such, well-worked out treatment protocols for the treatment of such lesions are in short supply. It appears as though mucosal melanomas (MuMs) develop more frequently in the nasal cavity and paranasal sinus region, and less often in the oral cavity. It seems that the incidence of nodal metastasis is significantly lower for sinonasal MuMs than it is for MuMs of the oral cavity; this observation may influence decisions about performing neck dissection as a function of location of the primary MuM. At present, surgical excision remains the mainstay of treatment; however, anatomical complexities within the region can hamper attempts at complete excision. Radiotherapy has not traditionally been relied on for routine treatment of MuM, although some recent reports have challenged this view. Chemotherapy is, at present, employed principally in the treatment of disseminated disease and for palliation. As a diagnostic matter, MuM belongs to the class of tumors that, on light microscopy, may with some regularity be confused with other malignancies (including sarcomas, plasmacytomas, and carcinomas); as a consequence, this is a diagnosis which is often best confirmed by way of ancillary testing via immunohistochemical studies. A better grasp of the best means of treating MuM will likely come only when large referral centers are able to pool their experiences with these uncommon yet virulent malignancies. PMID- 12772208 TI - In memoriam: Gabriel Ward Lasker (April 29, 1912-August 27, 2002). PMID- 12772209 TI - New approaches in hominoid taxonomy: morphometrics. AB - We report here on new cranial data relevant to hominoid taxonomic analyses, based on a study of 438 skulls belonging to 13 nonhuman living hominoid taxa. Nineteen landmarks were selected to describe the overall shape of the maxillofacial complex, in order to investigate its discriminative power in taxonomic analyses. We used a geometric morphometrics approach to depict morphological variation from the genus down to the subspecific level, and we evaluated whether our morphologic criteria are relevant to discriminating species and subspecies among living hominoids. Considering previous genetic studies, we discuss whether our results can be extrapolated to the hominin fossil record, providing a reference for species and subspecies morphologic differentiation. Our results indicate that the relative warp method, as applied to facial landmarks, provides a powerful tool to discriminate taxa down to a subspecific level. Results show a noticeable divergence of P. t. verus compared to P. t. troglodytes and P. t. schweinfurthii. According to our data, the distance between eastern and western gorilla populations as well as between Bornean and Sumatran orangutan subspecies is as great as between the two species of Pan. In the same manner, differences between Hylobates and Symphalangus are similar to those between Pan and Gorilla genera. Congruence between the morphological distances computed in this study and previous morphological and genetical studies strongly supports their relevance for morphological species recognition in paleoanthropology. Our data provide an objective standard for assessing taxonomic differences among hominoids, and will enable us to define more precisely the significance of morphological differences in the fossil record. PMID- 12772210 TI - Variation in ancient Egyptian stature and body proportions. AB - Stature and the pattern of body proportions were investigated in a series of six time-successive Egyptian populations in order to investigate the biological effects on human growth of the development and intensification of agriculture, and the formation of state-level social organization. Univariate analyses of variance were performed to assess differences between the sexes and among various time periods. Significant differences were found both in stature and in raw long bone length measurements between the early semipastoral population and the later intensive agricultural population. The size differences were greater in males than in females. This disparity is suggested to be due to greater male response to poor nutrition in the earlier populations, and with the increasing development of social hierarchy, males were being provisioned preferentially over females. Little change in body shape was found through time, suggesting that all body segments were varying in size in response to environmental and social conditions. The change found in body plan is suggested to be the result of the later groups having a more tropical (Nilotic) form than the preceding populations. PMID- 12772211 TI - Understanding muscle markers: aggregation and construct validity. AB - Musculoskeletal markers are frequently used to reconstruct past lifestyles and activity patterns. Yet, the reliability of muscle marker measurements has been called into question because they allegedly fail to correlate with cross sectional properties and exercise patterns, and are confounded by body size. In this study, the principle of aggregation was used to sum muscle markers over 7 insertion sites (4 humeral, 2 radial, and 1 ulnar) and examine the effects on them of body size, age, sex, and cross-sectional properties. Analyses were made of a sample of 91 (66 males, 25 females) Native British Columbians (3500-1500 years BP) and 18th century Quebec prisoners. Muscle markers were measured using three-point observer rating scales; size was measured by standard methods; age and sex were determined through pelvic, cranial, and dental morphology; and cross sectional properties were calculated from radiographs. Whereas any single muscle marker component failed to correlate with age, size, sex, or cross-sections, aggregate muscle marker correlated with: age, r = 0.49; size, r = 0.38; sex, r = 0.40; and, cross-sections, r = 0.38; P < 0.001. Older individuals had greater muscle markers, as did larger individuals, males, and those with more robust cross-sections. Based on partial correlations and regression analyses, age was the best overall predictor of aggregate muscle marker. PMID- 12772212 TI - Characterization of biological diversity through analysis of discrete cranial traits. AB - In the present study, the frequency distributions of 20 discrete cranial traits in 70 major human populations from around the world were analyzed. The principal coordinate and neighbor-joining analyses of Smith's mean measure of divergence (MMD), based on trait frequencies, indicate that 1). the clustering pattern is similar to those based on classic genetic markers, DNA polymorphisms, and craniometrics; 2). significant interregional separation and intraregional diversity are present in Subsaharan Africans; 3). clinal relationships exist among regional groups; 4). intraregional discontinuity exists in some populations inhabiting peripheral or isolated areas. For example, the Ainu are the most distinct outliers of the East Asian populations. These patterns suggest that founder effects, genetic drift, isolation, and population structure are the primary causes of regional variation in discrete cranial traits. Our results are compatible with a single origin for modern humans as well as the multiregional model, similar to the results of Relethford and Harpending ([1994] Am. J. Phys. Anthropol. 95:249-270). The results presented here provide additional measures of the morphological variation and diversification of modern human populations. PMID- 12772213 TI - Effect of parity on bone mineral density in female rhesus macaques from Cayo Santiago. AB - This cross-sectional study investigates the relationship between parity, bone mineral density, and spontaneous osteopenia/osteoporosis in a large skeletal population of female rhesus macaques (Macaca mulatta) from the free-ranging colony of Cayo Santiago, Puerto Rico. The sample consists of 119 mature female monkeys aged 4.0-22.2 years at time of death. The data consist of measurements of bone mineral content (BMC) and bone mineral density (BMD), obtained from dual energy X-ray absorptiometry (DEXA) of the last lumbar vertebra. After controlling for age, there is a significant increase in BMD of the spine with increasing parity (P = 0.0006), up to a parity of 7 offspring. Thus, high parity initially has a positive effect on BMD in female rhesus monkeys, but this positive effect disappears with parities that are greater than 7 offspring. After controlling for parity, however, age has a negative (P = 0.015) effect on BMD, beginning several years after the attainment of peak BMD (age 9.5 years). Thus, it appears that parity initially mitigates the effects of aging, but the positive effect of parity on BMD is eventually overwhelmed by the aging process. Mean BMC and BMD values are higher in parous females compared to nulliparous females in the same age range. Similarly, females with low parity have significantly lower mean BMD values than do age-matched high-parity controls, and the frequency of osteopenia and osteoporosis is greater in low-parity females. Forty-three percent (43%) of the osteopenic/osteoporotic females in the sample are members of the low-parity group, even though it composes only 13% (16/119) of the entire sample. This study demonstrates that the free-ranging female rhesus monkeys from Cayo Santiago are a good nonhuman primate model for the study of bone mineral density, parity, osteopenia, and osteoporosis. PMID- 12772214 TI - Peopling of three Mediterranean islands (Corsica, Sardinia, and Sicily) inferred by Y-chromosome biallelic variability. AB - An informative set of biallelic polymorphisms was used to study the structure of Y-chromosome variability in a sample from the Mediterranean islands of Corsica and Sicily, and compared with data on Sardinia to gain insights into the ethnogenesis of these island populations. The results were interpreted in a broader Mediterranean context by including in the analysis neighboring populations previously studied with the same methodology. All samples studied were enclosed in the comparable spectrum of European Y-chromosome variability. Pronounced differences were observed between the islands as well as in the percentages of haplotypes previously shown to have distinctive patterns of continental phylogeography. Approximately 60% of the Sicilian haplotypes are also prevalent in Southern Italy and Greece. Conversely, the Corsican sample had elevated levels of alternative haplotypes common in Northern Italy. Sardinia showed a haplotype ratio similar to that observed in Corsica, but with a remarkable difference in the presence of a lineage defined by marker M26, which approaches 35% in Sardinia but seems absent in Corsica. Although geographically adjacent, the data suggest different colonization histories and a minimal amount of recent gene flow between them. Our results identify possible ancestral continental sources of the various island populations and underscore the influence of founder effect and genetic drift. The Y-chromosome data are consistent with comparable mtDNA data at the RFLP haplogroup level of resolution, as well as linguistic and historic knowledge. PMID- 12772215 TI - The names of Spain: a study of the isonymy structure of Spain. AB - In order to estimate the isonymy structure of Spain, we studied surname distribution in 283 Spanish towns based on 3.625 million telephone users selected from 6.328 million users, downloaded from a commercial CD-ROM which contains all 13 million users in the country. Since in Spain the surname is made by the paternal and the maternal surname, it was possible to classify surnames according to parental origin. Two matrices of isonymy distances, one for paternal and one for maternal surnames, were constructed and tested for correlation with geographic distance. For the whole of Spain, Euclidean distance was significantly but weakly correlated with geographic distance both for paternal and maternal surnames, with r = 0.205 +/- 0.013 and r = 0.263 +/- 0.012, respectively. Two dendrograms of the 283 sampled towns were built from the two matrices of Euclidean distance. They are largely colinear. Four main clusters identified by the dendrograms are correlated with geography. Given the surname structure of Spain, we were able to calculate from isonymy and for each town 1). total or expressed inbreeding, 2). random or expected inbreeding, and 3). local inbreeding. Total inbreeding, F(IT), was highest in the North Atlantic regions and lowest along the Mediterranean Coast. The lowest levels were found in Andalusia, Catalunyia, Valencia, and Navarra. Random inbreeding, F(ST), had a similar geographical pattern. Local inbreeding, F(IS), was relatively uniform in the whole of Spain. In towns, random inbreeding dominates over local inbreeding. From the analysis, it emerges that the northwestern area of Spain is the most inbred. PMID- 12772217 TI - Nonclassic cystic fibrosis: a clinical conundrum. PMID- 12772216 TI - What is the role of tests of lung function in the management of infants with lung disease? AB - This review considers whether there is a role for lung function tests in the clinical management of infants with lung disease. The purpose of testing lung function in older subjects, the tests available for infants, and the practical problems of testing lung function in infants are considered. After reviewing all the facts, we suggest that there are four situations in which lung function testing should be recommended for infants, as follows: (1) the infant who presents with unexplained tachypnea, hypoxia, cough, or respiratory distress in whom a definitive diagnosis is not apparent from physical examination and other, less difficult investigations; (2) the infant with severe, continuous, chronic obstructive lung disease who does not respond to an adequate clinical trial of combined corticosteroid and bronchodilator therapy; (3) the infant with known respiratory disease of uncertain severity in whom there is need to justify management decisions; and (4) research and development. A review of 62 recent publications to determine how lung function tests are being used at the present time showed that they are being used overwhelmingly for research. The role of lung function testing in the clinical management of infants has not been established, and research is needed to clarify this situation. We suggest that such studies should explore the role of lung function tests in infants with specific symptoms, signs, or diagnoses, taking into account information from other types of investigation and the cost/benefit/risk ratios. PMID- 12772218 TI - Role of budesonide as maintenance therapy for children with asthma. AB - Asthma is a chronic inflammatory disease of the airways. Inhaled corticosteroids are recognized as the preferred long-term control medication for persistent asthma based on their anti-inflammatory properties and significant evidence of efficacy. Inhaled budesonide is the most carefully characterized inhaled corticosteroid for childhood asthma. It is available for administration in children down to six months of age and to date has an excellent safety profile. PMID- 12772219 TI - Hospital readmissions for asthma in children and young adults in Canada. AB - To examine the incidence rate of hospital readmission for asthma in relation to sex and age among Canadian children and young adults, we used data from 86,863 subjects under age 20 years when they had a first admission for asthma as 1 of first 5 diagnoses in Canada between April 1, 1994 and March 31, 1997. We calculated age- and sex-specific incidence rates, and used the Cox proportional hazards model for multivariate analysis. Of these subjects, 20,277 (23.3%) were readmitted to hospital for asthma during the study period. After adjusting for length of stay for first admission and province, the rate ratio for females vs. males was 0.86 for those under age 1 year, and close to unity for the 1-4-year and 5-9-year age groups, whereas it was 1.47 and 1.35 for the 10-14-year and 15 19-year age groups, respectively. The data showed similar trends for rehospitalization asthma as a primary diagnosis. The incidence rate of rehospitalization showed little sex difference between ages 1-9 years, but was markedly higher in females than in males 10-19 years of age. Airway size, female hormonal changes, increased use of cosmetic products, and cigarette smoking among adolescent girls may contribute to the age- and sex-differences in adolescence. PMID- 12772221 TI - Pulmonary gas trapping increases in asthmatic children and adolescents in the supine position. AB - This study aimed to see if gas trapping or ventilation inhomogeneity during tidal breathing increases in young asthmatic subjects when recumbent over a period of 1 hr. Ten asthmatic children and teenagers and 12 control subjects performed tidal breathing multiple-breath N(2)-washout in the sitting position and immediately, 30 min, and 60 min after assuming the supine position. The asthma group also performed the same lung function tests on another day, but then remained in the sitting position throughout. When assuming the supine position, both groups showed significant falls in vital capacity (4-8%) and in functional residual capacity (FRC) (>20%). Gas trapping increased significantly after 30 and 60 min in the supine position only in the asthma group, but other indices of ventilation inhomogeneity did not change with body position. When in the supine position, the trapped gas volume was on average 10% of FRC in the asthma group, which was twice as much as when sitting and three times higher than in supine control subjects. It is concluded that assuming the supine position leads to increased gas trapping in young asthmatics but not in healthy subjects; this could be one of several mechanisms that contribute to nocturnal worsening of asthma. PMID- 12772220 TI - Echocardiographic parameters in adolescents with sleep-related breathing disorders. AB - Few data are known about the effects of sleep-related breathing disorders (SRBD) on the cardiovascular system in adolescents. Forty healthy adolescents (mean age, 13.7 +/- 1.6 years) answered a questionnaire regarding symptoms of sleep-related breathing disorders (SRBD) and underwent a cardiorespiratory polygraphy and echocardiography. Echocardiographic data in snorers with polygraphic abnormalities suggestive of SRBD (positive group, n = 29) were compared with the results in nonsnorers without polygraphic abnormalities (n = 11) who were included in the control group. Ventricular dimensions and indices of left ventricular systolic function were within normal limits and were not significantly different between the two groups. Indices of left ventricular diastolic function were also within normal limits, but isovolumetric relaxation time (IVRT) was significantly longer among the positive group (72.5 +/- 8.4 msec) than among the controls (65.1 +/- 7.9 msec) (P = 0.018). Multiple regression analysis showed that posterior wall thickness was predicted by a model that included cardiac events related with respiratory events and/or desaturations, and respiratory disturbance index (RDI) in supine position. Deceleration of early diastolic flow was predicted by RDI, percentage of total recording time with SaO(2) < 90% (CT(90)), and age; the variability of isovolumetric relaxation time was predicted by a model that included RDI in supine position. We found a significant relationship between polygraphic parameters suggestive of SRBD and echocardiographic measurements of ventricular dimensions and diastolic function. Also, echocardiographic parameters suggestive of some degree of left ventricular diastolic dysfunction were found in snoring adolescents with polygraphic abnormalities. PMID- 12772222 TI - Duration of effect of intravenous antibiotics on spirometry and sputum cytokines in children with cystic fibrosis. AB - Intravenous (IV) antibiotics are a mainstay of therapy in children with cystic fibrosis. It is unclear, however, over what period associated improvements in pulmonary function are maintained, and to what extent the underlying inflammatory process is impeded in children admitted for a course of IV antibiotics. This was a prospective, interventional study of 14 children (median age, 14 years; interquartile range, 10-14) with cystic fibrosis who were regular sputum producers and who required admission for a 2-week course of IV antibiotics. Children performed spirometry and provided a sputum sample prior to starting IV antibiotics and then weekly for 6 weeks, the first 2 weeks of which IV antibiotics were given. Sputum IL-8, TNF-alpha, IL-6, IL-10, MIP1-alpha, and elastase were measured. Seven children were asked to repeat the protocol in a subsequent exacerbation to assess similarities in response to therapy. Significant improvements were seen in forced expired volume in 1 sec (FEV(1)) in association with IV antibiotics (27% relative improvement in predicted from baseline to end of week 1, median FEV(1) 41.3% increasing to 52.2%), but this continued only 1 week following cessation of antibiotics. Although IL-8 demonstrated a trend for reduction in association with antibiotics, no significant profile was demonstrated for any of the cytokines assessed. IL-10 was detectable in 64% of samples (all <100 pg/ml). In children with two episodes assessed, although there was a close correlation of FEV(1) and FVC between exacerbations (before antibiotics), no significant correlation was seen for IL-8, TNF-alpha, or IL-10 measured in both sets of samples at any sample point (indeed, a discordant response was seen between sample points in the two exacerbations). Although FEV(1) temporarily improves in response to admission for IV antibiotics, no such response is seen in sputum cytokine values. In addition, assessment of cytokines in subsequent exacerbations does not show a similar pattern of response to treatment. PMID- 12772223 TI - Exercise-induced bronchospasm in children: comparison of FEV1 and FEF25-75% responses. AB - The response of asthmatic children to exercise has usually been evaluated by forced expiratory volume in 1 sec (FEV(1)). We reasoned that other respiratory indexes derived from the forced vital capacity maneuver such as forced expiratory flow between 25-75% of vital capacity (FEF(25-75%)) would add significant information in the evaluation of the relationship between asthma severity and response to exercise. We studied 164 children with intermittent (n = 63), mild persistent (n = 30), moderate persistent (n = 40), and severe persistent asthma (n = 31). Subjects exercised for 6 min on a cycle ergometer at 80% of their maximum heart rate, and spirometry was performed before and 5, 10, and 20 min after exercise. There was good correlation between changes in FEV(1) and FEF(25 75%) after exercise (r = 0.60, P < 0.001 for intermittent asthma and r = 0.80, P < 0.001 for severe persistent asthma). The presence of a fall in both FEV(1) (>/=10%) and in FEF(25-75%) (>/=26%) when compared to a decrease in only one of these two indexes was significantly greater in children with more severe asthma (60.0% for intermittent asthma and 94.4% for severe persistent asthma, P = 0.022). FEF(25-75%) can decrease in response to exercise without changes in FEV(1), mainly in children with mild asthma. In the evaluation of the response to exercise in children with different asthma severities, more than one maximum expiratory flow-volume parameter should be used. PMID- 12772224 TI - Follow-up of compliance with tuberculosis treatment in children: monitoring by urine tests. AB - This study was designed to follow up patient compliance by detection of antituberculous drugs in urine during the course of treatment. It was conducted in the Outpatient Clinic of Pediatric Infectious Diseases, Sisli Etfal Hospital (Istanbul, Turkey). In total, 45 children with pulmonary tuberculosis participated. Patients were seen twice in the first month and once a month thereafter during the 6-month course of treatment. The second urine of the day was collected at each visit. Urine was tested for isoniazid (INH), rifampicin (RIF), and pyrazinamide (PZA). In the presence of these drugs or their metabolites, the addition of certain chemicals caused a color change in the urine. On day 15 of treatment, urine tested positive for INH in 82% of patients, for RIF in 67%, and for PZA in 73%. At the end of the second month, the ratio of adherence was 96, 89, and 96% for each drug, respectively. All patients were found to be adherent at months 5 and 6. We recommend detection of antituberculous drugs in urine to assess compliance to treatment. Once the defaulting patients were identified, adherence was improved by repeatedly providing patient education throughout the treatment. PMID- 12772225 TI - Sputum induction as a diagnostic tool for community-acquired pneumonia in infants and young children from a high HIV prevalence area. AB - Sputum induction is a standard diagnostic procedure to identify pathogens in lower respiratory tract secretions in adults with pneumonia, but has rarely been studied or used in infants and young children. Our aim was to determine the usefulness of induced sputum (IS) as a diagnostic method for infants and children hospitalized with community-acquired pneumonia (CAP) in a high HIV prevalence area. Children hospitalized for CAP were prospectively enrolled over a year. IS was obtained by nebulization with hypertonic (5%) saline, physiotherapy, and suctioning. Sputum was submitted for bacterial and mycobacterial culture and P. carinii detection. Gastric lavages (GLs) were done for M. tuberculosis culture; a nasopharyngeal aspirate (NPA) was obtained for bacterial culture and P. carinii detection. IS was obtained in 210 children (median age, 7 (25th to 75th percentile, 3-18) months); 138 (66%) were HIV-infected; 148 (70%) were receiving supplemental oxygen. Bacteria were isolated from 101 (50%) IS and 141 (70%) NPA paired specimens (P < 0.001). A significantly higher rate of S. aureus, H. influenzae, M. catarrhalis, and S. pneumoniae was found in NPAs compared to IS; this pattern was particularly evident in HIV-infected children. M. tuberculosis was cultured from sputum in 19 patients (9%); GLs performed in 142 children were positive in only 9 (6%). The difference (95% confidence interval) between yields for M. tuberculosis from culture of IS compared to GL was 4.3% (95% CI, 0-5.6%; P = 0.08). P. carinii was identified from IS in 12 (5.7%) children; all corresponding NPAs were negative. Seven (3%) children could not tolerate sputum induction. Side effects included increased coughing in 4%, epistaxis in 3%, and wheezing responsive to bronchodilators in 1%. In conclusion, induced sputum is a useful and safe diagnostic procedure in infants and children with CAP from a high HIV prevalence area. PMID- 12772226 TI - Majority of children aged 3 years and above can reliably inhale through the Clickhaler. AB - Guidelines suggest that pressurized metered dose inhalers (pMDI) plus spacers are the delivery system of choice for children. However, they are bulky, which makes them inconvenient. It was suggested that the smaller dry-powder inhalers (DPIs) may be suitable for delivering short-acting bronchodilators to preschool children. This study considered whether preschool children could reliably generate sufficient inspiratory flows to use the Clickhaler DPI. Twenty-seven asthmatic and 34 nonasthmatic children, aged 2-5 years, were recruited. Following training, they were asked to inhale four times through a Clickhaler flow monitoring system, twice "steadily" and twice "forcefully." Inspiratory flow data were collected during each inhalation. Of the 3-, 4-, and 5-year-old asthmatics, 62.5, 100, and 100%, respectively, could reliably differentiate between inhaling and exhaling through the DPI. For nonasthmatics, the figures were 66, 60, and 88%, respectively. All but one of the children who understood the instructions generated a PIF of greater than 15 l/min (range, 13.9-88.3 l/min and 21.2-84.5 l/min in asthmatic and nonasthmatic children, respectively). These data indicate that the majority of children aged 3 years and above could reliably inhale rather than exhale through a dry-powder inhaler, and that they generate inspiratory flows sufficient to use the Clickhaler effectively. The results indicate that the device could be a suitable delivery system for beta(2)-agonists in preschool children. PMID- 12772227 TI - Familial neonatal pneumothorax associated with transient tachypnea of the newborn. AB - We describe neonatal spontaneous pneumothorax associated with transient tachypnea of the newborn in siblings of two families. Familial spontaneous pneumothorax is extremely rare in neonates. Was our observation just an incidental finding, or is there a familial predisposition to spontaneous pneumothorax? PMID- 12772228 TI - Psychogenic cough in a schoolboy: evaluation using an ambulatory cough recorder. AB - Psychogenic cough occurs most commonly in patients under 18 years of age. Making the diagnosis on clinical features alone is problematic, and it is usually a diagnosis of exclusion after several negative clinical investigations. We report on the case of a 13-year-old schoolboy with a 3-month history of persistent dry cough with no other associated symptoms. Clinical examination and investigations revealed no abnormality, and empirical trials of antiasthma and antacid medications proved unsuccessful. An objective assessment of his cough frequency was made using an ambulatory cough monitor. A large number of cough episodes were recorded during the day, but during the time he was in bed there were very few episodes recorded. This suggested a diagnosis of psychogenic cough, and he underwent behavior modification therapy under the guidance of a clinical psychologist, with good result. Objective cough monitoring may therefore improve the evaluation and management of chronic cough. PMID- 12772229 TI - Granulomatous pulmonary disease in a child: an unusual presentation of Crohn's disease. AB - Crohn's disease (CD) is a chronic inflammatory disorder of the bowel which may be associated with an extensive list of extraintestinal manifestations involving almost every organ system. The most common organs involved are the eyes, skin, joints, and liver. Symptomatic bronchopulmonary disorders have been reported only rarely in pediatric CD. We report on an 11-year-old child who had a recurrent cough and increasing dyspnea with exercise for 8 months before developing any gastrointestinal symptoms. He was demonstrated to have granulomatous inflammation of the lung, as well as of the gastrointestinal tract. Similarities between CD and sarcoidosis are discussed. PMID- 12772230 TI - Subcutaneous emphysema secondary to foreign-body aspiration. AB - Pneumomediastinum is uncommon in pediatric medical practice, outside the neonatal period. While asthma or respiratory infections are the most frequent underlying causes, it is important not to forget the possibility of foreign body aspiration, particularly after the clinical presentation. PMID- 12772231 TI - Trained and supervised physician assistants can safely perform diagnostic cardiac catheterization with coronary angiography. AB - Using a prospectively collected database of patients undergoing cardiac catheterization, we sought to compare the outcomes of procedures performed by supervised physician assistants (PAs) with those performed by supervised cardiology fellows-in-training. Outcome measures included procedural length, fluoroscopy use, volume of contrast media, and complications including myocardial infarction, stroke, arrhythmia requiring defibrillation or pacemaker placement, pulmonary edema requiring intubation, and vascular complications. Class 3 and 4 congestive heart failure was more common in patients who underwent procedures by fellows compared with those undergoing procedures by PAs (P = 0.001). PA cases tended to be slightly faster (P = 0.05) with less fluoroscopic time (P < 0.001). The incidence of major complications within 24 hr of the procedure was similar between the two groups (0.54% in PA cases and 0.58% in fellow cases). Under the supervision of experienced attending cardiologists, trained PAs can perform diagnostic cardiac catheterization, including coronary angiography, with complication rates similar to those of cardiology fellows-in-training. PMID- 12772232 TI - Hydrophilic coating aids radial sheath withdrawal and reduces patient discomfort following transradial coronary intervention: a randomized double-blind comparison of coated and uncoated sheaths. AB - Radial artery spasm may cause severe discomfort during radial artery sheath removal. A hydrophilic-coated sheath may reduce the force required to remove a radial sheath. This force may be quantified using an automatic pullback device (APD). The objective of this study was to assess if a hydrophilic coating reduces the required force and discomfort associated with removal of a radial sheath following transradial coronary intervention. Ninety patients undergoing percutaneous coronary intervention via the radial artery were randomly assigned to two groups receiving either coated or uncoated introducer sheaths. Radifocus Introducer II (Terumo) 25 cm, 6 Fr radial sheaths and sheaths that were identical apart from the presence of the coating were used in all patients. The APD was used for sheath removal at the end of the procedure. Three patients (7%) in the coated group experienced discomfort during automatic sheath removal, compared to 12 patients (27%) in the uncoated group (P = 0.02). The maximum pullback force (MPF) was significantly lower in the coated compared to the uncoated group (0.24 +/- 0.31 vs. 0.44 +/- 0.33 kg; P = 0.003). Similarly, the mean pullback force was significantly lower in the coated group (0.14 +/- 0.23 vs. 0.32 +/- 0.24 kg; P < 0.001). Only one patient (2%) in each group had an MPF greater than 1.0 kg together with clinical evidence of radial artery spasm. Removal of the coated Terumo Radifocus sheath requires less force than an identical uncoated sheath. The coated sheath was also associated with less discomfort for the patient. PMID- 12772233 TI - Vessel caliber and restenosis: a prospective clinical and angiographic study of NIR stent deployment in small and large coronary arteries in the same patient. AB - Retrospective analyses of patient cohorts undergoing stent deployment have shown that small vessel diameter and long lesion length are two angiographic predictors of increased restenosis. We determined the effects of these factors in patients with lesions treated in both small- and large-diameter coronary arteries. This multicenter prospective quantitative angiographic study evaluated patients with de novo coronary disease undergoing intervention who had at least two lesions < or = 16 mm length, one in a vessel < or = 2.75 mm diameter (9 or 16 mm length seven-cell NIR stent) and the other in a vessel > or = 3.0 mm diameter (9 or 16 mm nine-cell NIR stent). Of 94 patients enrolled, 76% were male, mean age was 62 years (range, 40-85), 41% were hypertensive, 18% had diabetes, 15% were current smokers, and 64% had hypercholesterolemia. Additional lesions were treated in 23% of patients. The procedural success rate was 99%. Six months postprocedure, there were no deaths or late stent occlusions. One patient suffered a Q-wave myocardial infarction, one a non-Q-wave infarction, eight underwent percutaneous reintervention, two coronary artery bypass graft surgery operations, and five stenting of other nonstudy lesions. The mean reference diameter for the small vessel was 2.35 mm and the large vessel 3.22 mm. Six-month angiography was performed in 87 patients (92% of those eligible). The overall restenosis rate was 24% in the small vessel (9 mm length stent, 17%; 16 mm length stent, 30%) and 15% in the large vessel (9 mm length stent, 3%; 16 mm length stent, 22%), respectively. Multivessel stenting including treatment of lesions in small caliber vessels can be performed with a good clinical and angiographic outcome. When the patient, operator, technique, and stent type are the same, vessel caliber and stent length both appear to influence the restenosis rate. PMID- 12772234 TI - Percutaneous interventions in radial artery grafts: clinical and angiographic outcomes. AB - In the modern era, radial artery graft is being used with increasing frequency to replace saphenous vein as a conduit for coronary artery bypass surgery. Several reports have shown encouraging early results of radial grafts compared to saphenous grafts. Despite these advantages, radial artery graft failure requiring revascularization does occur. We report on the clinical, angiographic, and technical characteristics and the follow-up results of 22 patients who underwent percutaneous intervention of radial grafts. PMID- 12772236 TI - Is adjunctive balloon postdilatation necessary after coronary stent deployment? Final results from the POSTIT trial. AB - Early-generation balloon-expandable stents required postdilatation with noncompliant balloons at high pressure to optimize stent deployment. The need for adjunctive balloon postdilatation with modern stent delivery systems is unknown. Patients undergoing elective stenting were randomized to Boston Scientific NIR, Guidant Tri-Star/Tetra, and Medtronic AVE S670 stents. The primary endpoint was optimum stent deployment defined as a minimal stent diameter (MSD) >/= 90% of the average reference lumen diameter assessed by intravascular ultrasound (IVUS) performed immediately following stent deployment. If, by operator assessment, the primary endpoint was not achieved with the stent delivery system, adjunctive postdilatation with noncompliant balloons was performed. Of 256 patients with IVUS studies adequate for core laboratory analysis, only 29% achieved optimum stent deployment with the stent delivery system. None of the baseline clinical or angiographic variables predicted optimum stent deployment. Of the procedural variables, the type of stent and nominal stent size were not predictors, but higher deployment pressures were associated with a higher frequency of optimum stent deployment (< 12 atm 14% vs. >/= 12 atm 36%; P = 0.007). The inability to achieve optimum stent deployment was not due to undersizing the stent delivery balloon, but rather to an inability of the stent delivery balloon to expand fully the stent to nominal size. In patients who underwent postdilatation, the frequency of achieving optimum stent deployment increased from 21% to 42%, minimal stent area increased from 6.6 +/- 2.2 to 7.8 +/- 2.3 mm(2), and MSD increased from 2.6 +/- 0.5 to 2.8 +/- 0.4 mm. These data stress the continued need for adjunctive balloon postdilatation with modern stent delivery systems. Cathet Cardiovasc Intervent 2003;59:184-192. PMID- 12772235 TI - Correlates of failure following treatment with Sr-90 beta irradiation for in stent restenosis. AB - We sought to determine the correlates of failure following intracoronary radiation therapy (IRT) with Sr-90 using the Novoste Beta-Cath system for the treatment of in-stent restenosis (ISR) in a broad range of patients. IRT has been shown to be more efficacious compared to placebo for the treatment of ISR in large randomized trials. However, even in patients treated with IRT, major adverse cardiac events occur in approximately 20% of cases on follow-up. This trial sought to elucidate the correlates of failure following successful IRT for ISR. To determine the correlates of IRT failure, we retrospectively compared the demographics, lesion characteristics, and clinical outcomes of 102 consecutive patients with ISR treated with Sr-90 from September 1998 to July 2001. IRT failure was defined as death, myocardial infarction (MI), or target vessel revascularization (TVR) due to repeat ISR on follow-up. A comparison of the clinical and angiographic profile of IRT failures (n = 16) vs. IRT successes (n = 86) revealed that a history of smoking (75% vs. 40%; P = 0.012), current use of calcium channel blockers (84% vs. 45%; P = 0.013), ostial location of target lesion (44% vs. 16%; P = 0.020), and mean posttreatment minimal luminal diameter (MLD; 1.64 +/- 0.19 vs. 2.21 +/- 0.29 mm; P < 0.001), respectively, were correlated with failure using univariate analysis. After multivariate regression analysis, the correlates of failure that remained significant were treatment of an ostial lesion (OR = 31.2; 95% CI = 2.6-382.7; P = 0.007) and final posttreatment MLD (P < 0.001). Ostial location of target lesion and smaller posttreatment MLD are correlated with subsequent death, MI, and TVR following therapy with Sr-90 for ISR. PMID- 12772237 TI - Three-two-one? PMID- 12772238 TI - The dilemma of success: percutaneous coronary interventions in patients > or = 75 years of age-successful but associated with higher vascular complications and cardiac mortality. AB - Elderly patients are increasingly referred to percutaneous coronary interventions (PCIs). Recent reports suggest complications rates are declining in the elderly. We sought to determine whether procedural and in-hospital outcomes are different in patients aged > or = 75 years undergoing nonemergent PCI as compared to patients age < 75 years. The outcome of 266 consecutive patients age > or = 75 years undergoing nonemergent PCI was compared to that of 1,681 consecutive patients age < 75 years. Compared with younger patients, greater proportions of elderly patients were women and had a history of hypertension, peripheral vascular disease, and cerebral vascular events. Elderly patients had more extensive coronary involvement. Procedural success was similar in both groups (94%). The in-hospital cardiac death rate was significantly higher in the elderly patients (2.3% vs. 0.7%; P = 0.03). Aged patients also had a significantly higher incidence of vascular and bleeding complications. Blood transfusion was required more often in the elderly group (4.5% vs. 2.6%; P = 0.07). The hospitalization length was significantly higher in the elderly group (4.1 +/- 6.0 vs. 2.5 +/- 4.3 day; P = 0.0004). By multivariate logistic regression (adjusted for baseline clinical and angiographic variables), age > or = 75 years was found to be an independent predictor of in-hospital cardiac death (odds ratio = 3.9; 95% CI = 1.3-11.5; P = 0.015). Although PCI is technically successful in patients aged > or = 75 years; it is associated with more acute cardiac and vascular complications and higher in-hospital cardiac mortality. PMID- 12772239 TI - Percutaneous coronary interventions in the elderly: Just the tip of the iceberg. PMID- 12772240 TI - Endovascular hypothermia in the setting of myocardial infarction complicated by cardiac arrest: a case report. PMID- 12772241 TI - Nonhealing wound resulting from a foreign-body reaction to a radial arterial sheath. AB - Several patients developed sterile inflammation at their radial arterial access site. Pathologic examination of biopsy material from one patient demonstrated a foreign-body reaction to material most likely from the gel-coated arterial access sheath. Surgical excision of the inflamed tissue resulted in healing. PMID- 12772242 TI - Sterile inflammation associated with transradial catheterization and hydrophilic sheaths. AB - In 1999, we noted the development of inflammation and/or abscesses at the site of radial access in a group of patients. Over a 3-year period, we noted this inflammation in 33 patients out of 2,038 (1.6%) who had catheterization via the radial approach. The radial abscesses occurred in 30 patients out of 1,063 (2.8%) in whom we could confirm the use of a hydrophilic-coated sheath, but in no patient for whom we can document that an uncoated sheath was used. No infectious agent could be implicated, and the time course for the development of the abscess, typically 2 to 3 weeks, seemed long for a bacterial infection. Later patients had biopsies, and granulomatous reactions were seen in most. Additionally, a few of the biopsies showed an amorphous extravascular substance consistent with the catheter coating. All patients had good long-term outcomes. PMID- 12772243 TI - Catheter-induced left main dissection: a treatment dilemma. AB - Diagnostic angiography is a routine diagnostic procedure in modern day cardiological practice. However, there are potential life threatening complications which can occasionally occur. We describe one such complication and detail the management and clinical outcome. PMID- 12772244 TI - Left main dissection: a time bomb? PMID- 12772245 TI - Rapid thrombectomy for treatment of macroembolization during percutaneous coronary intervention in the setting of acute myocardial infarction. AB - We report the use of the Export catheter as an urgent modality to aspirate thrombus that embolized down the left anterior descending artery during acute myocardial infarction. PMID- 12772246 TI - Recanalization of chronic and long occlusive in-stent restenosis using optical coherence reflectometry-guided radiofrequency ablation guidewire. AB - Chronic total occlusions remain a challenge to interventionalists due to failure of crossing or perforation by strong wires. We describe the use of a system using optical coherence reflectometry for navigation and radiofrequency ablation to enable wire passage through occlusions that could not be crossed by conventional guidewires. PMID- 12772248 TI - Closure of postmyocardial infarction VSD: a call for new devices. PMID- 12772247 TI - Compassionate use of the amplatzer ASD closure device for residual postinfarction ventricular septal rupture following surgical repair. AB - We report successful transcatheter closure of a post-MI ventricular septal rupture acutely following unsuccessful surgical repair. Catheter closure was accomplished by the use of a 26-mm Amplatzer atrial septal occluder. Initial attempts to close the defect with the use of 28-mm and 33-mm CARDIOSEAL were unsuccessful. Closure technique, immediate and long-term follow-up outcomes are reported. PMID- 12772249 TI - Primary angioplasty in CADASIL. AB - We report an acute coronary event in a young patient suffering from cerebral autosomal dominant arteriopathy with sub-cortical infarcts and leucoencephalopathy (CADASIL) successfully treated by primary angioplasty and direct stenting. Coronary involvement and pathophysiology of this genetic disorder is discussed. PMID- 12772250 TI - Transcatheter closure of recurrent postmyocardial infarction ventricular septal defects utilizing the Amplatzer postinfarction VSD device: a case series. AB - The initial therapy for postmyocardial infarction ventricular septal defects is surgical repair of the defect. Unfortunately, a significant number of patients develop recurrent ventricular septal defects (VSDs) following operative repair. Transcatheter closure offers an alternative to reoperation in these critically ill patients. We present a series of four patients in whom recurrent ventricular septal defects were closed using an Amplatzer VSD device. PMID- 12772251 TI - Unusual vertebral artery origins: examples and related pathology. AB - Anomalies of the vertebral arteries are uncommon, but important to recognize in the diagnosis and catheter based evaluation and treatment of patients suffering cerebrovascular disease. This article illustrates our experience with such anomalies. These include the vertebral artery arising as the fourth and most distal branch of the aortic arch, as a right subclavian artery branch arising distal to the right thyrocervical trunk, as a right common carotid artery branch in a patient with an aberrant right subclavian artery, and a case of left vertebral artery proximal duplication, with both aortic and left subclavian vertebral arteries present in the same patient; the latter join to form a single distal cervical vertebral artery. PMID- 12772252 TI - Endovascular brachytherapy for treatment of bilateral renal artery in-stent restenosis. AB - Percutaneous transluminal angioplasty of renal artery stenosis is an attractive alternative to surgical therapy. However, even with endovascular stenting, the overall rate of restenosis is 21%. While brachytherapy for coronary in-stent restenosis has proven efficacy, its use for renal artery in-stent restenosis has not been formally evaluated. We report a case of bilateral in-stent renal artery restenosis treated with endovascular brachytherapy. PMID- 12772253 TI - First case reports of controlled blunt microdissection for percutaneous transluminal angioplasty of chronic total occlusions in peripheral arteries. AB - Percutaneous transluminal angioplasty (PTA) can fail to revascularize peripheral arteries when a chronic total occlusion (CTO) cannot be crossed by guidewires. This article describes application of a new controlled blunt microdissection (CMD) catheter designed to cross CTOs. Two men presenting with severe claudication had iliac CTOs that resisted crossing with guidewires. Using standard techniques, the CMD catheter was advanced to the CTO. Following attempts to cross the CTO with guidewires, the jaw of the CMD distal assembly was actuated, advancing through the CTO as plaque was blunt-dissected. After angioplasty and stenting, restored distal flow was restored. Ischemic symptoms had not recurred at 1- and 28-month follow-up. The concept of blunt intraluminal microdissection has been applied to convert failing to successful PTA of peripheral arteries. CTOs that had resisted guidewire crossing were successfully crossed using the CMD catheter, allowing treatment by angioplasty and stenting. PMID- 12772254 TI - Total occlusion: old vs. new technology. PMID- 12772255 TI - Transjugular percutaneous inoue balloon mitral commissurotomy in a patient with inferior vena cava obstruction after liver transplantation. AB - Percutaneous transvenous mitral commissurotomy was performed successfully via the transjugular approach in a patient with severe rheumatic mitral stenosis and obstruction of the inferior vena cava due to prior liver transplantation. This case demonstrates the advantage of the jugular approach in patients with difficult anatomy. PMID- 12772256 TI - All roads lead to Rome: transjugular or transfemoral approach to percutaneous transseptal balloon mitral valvuloplasty? PMID- 12772257 TI - Iliofemoral arterial malformation associated with single umbilical artery: implications for interventional catheterization? PMID- 12772258 TI - One-step treatment of patent ductus arteriosus and pulmonary artery stenosis by cardiac catheterization. AB - Coexistence of pulmonary artery (PA) stenosis and patent ductus arteriosus (PDA) may lead to overestimating the relative importance of these malformations. This article reports on a patient with a trivial PDA and severe PA stenosis erroneously diagnosed as moderate PDA. He was successfully submitted to simultaneous PA stenting and PDA embolization. PMID- 12772259 TI - A one-step treatment and avoidance of a misstep. PMID- 12772260 TI - Invasive echocardiography: the use of catheter imaging by the interventional cardiologist. AB - Ultrasound imaging is frequently used for diagnostic purposes or guidance during procedures in the pediatric and congenital cardiac catheterization laboratory. As new imaging modalities emerged, many interventional cardiologists rather than noninvasive specialists are now performing the ultrasound imaging as part of the catheterization. The focus of this discussion will be to detail the technique and application of echocardiography by the interventional cardiologist. PMID- 12772261 TI - Anterograde balloon aortic valvuloplasty in the neonate via the umbilical vein. PMID- 12772262 TI - Use of a guidewire circuit in balloon valvuloplasty of the aortic valve. PMID- 12772263 TI - The patient. PMID- 12772264 TI - Isolation of ammonium-N as 1-sulfonato-iso-indole for measurement of delta15N. AB - The conversion of ammonium (NH(4) (+)) to 1-sulfonato-iso-indole has been examined as a method for natural abundance measurement of delta(15)N of NH(4) (+). The reaction is complete within 2 h and is based on the derivatisation of NH(4) (+) by o-phthaldialdehyde and sodium sulfite at a high pH, 11.2. The product is readily concentrated from dilute solutions by reverse-phase solid phase extraction (SPE). The method is compound-specific despite partial derivatisation of potentially interfering amino acids, as their derivatives are not extracted by SPE. delta(15)N values of NH(4) (+) in KCL soil extracts can be measured within 48 h by automated continuous-flow IRMS with a precision of 0.23 per thousand (1 SD). Parallel measurements of NH(4) (+) standards of known delta(15)N are made to allow correction for the isotopic dilution by non-sample NH(4) (+). The practicality of this method is demonstrated by measuring the changes in NH(4) (+) concentration and delta(15)N following the addition of urea as a nitrogen source to inorganic N-depleted soil. PMID- 12772265 TI - Sample preparation effects in matrix-assisted laser desorption/ionisation time-of flight mass spectrometry of partially depolymerised carboxymethyl cellulose. AB - Sample preparation effects in matrix-assisted laser desorption/ionisation time-of flight mass spectrometry (MALDI-TOFMS) of partially depolymerised carboxymethyl cellulose (CMC) have been investigated. The depolymerisation was either enzymatic or acidic. Fractions of enzymatically depolymerised CMC were collected from size exclusion chromatography (SEC) and further investigated by MALDI-TOFMS. 2,5 Dihydroxybenzoic acid was used as matrix, dissolved in H(2)O due to the poor solubility of CMC in suitable organic solvents. The samples were dried by two methods, in ambient atmosphere and at reduced pressure. Under reduced pressure the sample spot homogeneity increased. This drying method, however, produced additional adduct peaks in the mass spectra originating from ion exchange on the CMC oligomers. Analysis of CMC could be performed in both negative and positive ion modes. Mass discrimination and variation in ionisation efficiency were demonstrated by comparing mass spectra with SEC data. Measurements of the degree of substitution (DS) were performed on three CMCs with different DS values, which were depolymerised in trifluoroacetic acid. The three CMCs were easily distinguished from one another, but the obtained DS values deviated from the values supplied by the manufacturer. PMID- 12772266 TI - Sample preparation effects in matrix-assisted laser desorption/ionisation time-of flight mass spectrometry of partially depolymerised methyl cellulose. AB - Methyl cellulose (MC) was partially depolymerised and the oligomers thus obtained were studied by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOFMS). The depolymerisation was either enzymatic or acidic. Fractions of enzymatically depolymerised MC were collected from size-exclusion chromatography and subjected to a sample preparation investigation. Several MALDI matrices and solvents were evaluated. The results showed that the solvent choice had a significant effect on the measured degree of substitution (DS). Aprotic solvents produced higher DS values, which was most likely due to poor solubility of species with low DS. The obtained signal intensity, however, did not correlate with the solubility but seemed to be more dependent on certain matrix/solvent combinations. All the matrices attempted produced mass spectra with sufficient signal intensity for accurate peak area calculation. The choice of matrix did not have any significant effect on the measured DS. Sample spots obtained from organic solvents had a more homogeneous distribution of the analyte and smaller crystals than those obtained from water. This increased both the reproducibility and peak resolution and in addition the analysis time was shorter. DS measurements were performed on two acidically depolymerised MCs with different nominal DS values. It was easy to distinguish between the two MCs, and the measured DS values agreed well with the values supplied by the manufacturers. PMID- 12772268 TI - Analysis of urinary nucleosides. III. Identification of 5'-deoxycytidine in urine of a patient with head and neck cancer. AB - Modified nucleosides in the urine have been postulated to be diagnostic indices of disease, particularly of cancer. The urine of a patient with terminal head and neck cancer has been found to contain a modified nucleoside with a protonated molecule of m/z 228. By means of high-performance liquid chromatography/ion trap mass spectrometry (HPLC/ITMS) and capillary liquid chromatography/triple quadruple mass spectrometry (CapLC/TQMS) we have identified the compound as 5' deoxycytidine. This is the first report of 5'-deoxycytidine in man: in addition to the elucidation of its structure, its possible origins and the potential significance of its occurrence are discussed. PMID- 12772267 TI - Identification of diastereomeric chlorophyll allomers by atmospheric pressure chemical ionisation liquid chromatography/tandem mass spectrometry. AB - Atmospheric pressure chemical ionisation liquid chromatography/mass spectrometry (APCI-LC/MS) has been used for identification of the epimers of hydroxy, methoxy and methoxylactone allomers of chlorophyll a (13(2)-HO-chl a, 13(2)-MeO-chl a and 15(1)-MeO-lact-chl a), the hydroxy allomer of bacteriochlorophyll a (13(2)-HO bchl a) and the hydroxy and methoxylactone allomers of bacterioviridin a (13(2) HO-bvir a and 15(1)-MeO-lact-bvir a). The APCI mass spectra show that facile fragmentations involve the methoxyl or hydroxyl groups at the C-13(2) or C-15(1) chiral centres. Losses involving the C-13(2) or C-15(1) hydroxyl or methoxyl groups occur more easily from the S-epimer than from the R-epimer due to the greater relief of the steric strain associated with interaction with the bulky C 17 substituent. The differences in mass spectrometric fragmentation can be used as a diagnostic tool for the assignment of the stereochemical configuration at the C-13(2) or C-15(1) chiral centres. PMID- 12772269 TI - Use of a quadrupole linear ion trap mass spectrometer in metabolite identification and bioanalysis. AB - A new type of quadrupole linear ion trap mass spectrometer, Q TRAP trade mark LC/MS/MS system (Q TRAP trade mark ), was evaluated for its performance in two studies: firstly, the in vitro metabolism of gemfibrozil in human liver microsomes, and, secondly, the quantification of propranolol in rat plasma. With the built-in information-dependent-acquisition (IDA) software, the instrument utilizes full scan MS in the ion trap mode and/or constant neutral loss scans as survey scans to trigger product ion scan (MS(2)) and MS(3) experiments to obtain structural information of drug metabolites 'on-the-fly'. Using this approach, five metabolites of gemfibrozil were detected in a single injection. This instrument combines some of the unique features of a triple quadrupole mass spectrometer, such as constant neutral loss scan, precursor ion scan and multiple reaction monitoring (MRM), together with the capability of a three-dimensional ion trap. Therefore, it becomes a powerful instrument for metabolite identification. The fast duty cycle in the ion trap mode allows the use of full product ion scan for quantification. For the quantification of propranolol, both MRM mode and full product ion scan in the ion trap mode were employed. Similar sensitivity, reproducibility and linearity values were established using these two approaches. The use of the product ion scan mode for quantification provided a convenient tool in selecting transitions for improving selectivity during the method development stage. PMID- 12772270 TI - Confirmatory analysis of sulfonamide antibacterials in bovine liver and kidney: extraction with hot water and liquid chromatography coupled to a single- or triple-quadrupole mass spectrometer. AB - A simple, specific, and rapid confirmatory method for determining 12 sulfonamide (SAs) antibacterials in bovine liver and kidney is presented. This method is based on the matrix solid-phase dispersion technique with hot water as extractant followed by liquid chromatography/mass spectrometry (LC/MS) with an electrospray ion source. The method was tailored for use with both single-quadrupole MS (I) and triple-quadrupole MS (II) instruments. After acidification and filtration of the aqueous extract, a 250-microL aliquot was injected into instrument I while only 25 microL was analyzed by instrument II. With instrument I MS data acquisition was performed in the selected ion monitoring (SIM) mode, selecting at least three ions for each target compound. With instrument II the selected reaction monitoring (SRM) mode with three fragmentation reactions for each compound was chosen. With the exception of sulfaquinoxaline (SQX), recovery of the analytes at the 50 ppb level in both liver and kidney was 72-96% with relative standard deviations (RSDs) ranging between 3 and 11%. The very poor recovery of SQX was due to its rapid enzymatic oxidation when in contact with the two tissues. With instrument I, limits of quantification (LOQs, S/N = 10) were 5 14 ppb of SAs. Even lower LOQs (1-8 ppb) were estimated by using instrument II, even though the extract volume analyzed was ten times lower than that with instrument I. With both matrices and using instrument I, severe ion signal suppression was experienced for the early-eluted SAs when trying to fractionate analytes by using a short chromatographic run time. This effect was traced to polar endogenous co-extractives eluted in the first part of the chromatographic run that interfered with gas-phase ion formation for SAs. Adopting more selective chromatographic conditions minimized this effect. PMID- 12772271 TI - Simultaneous quantitation of enalapril and enalaprilat in human plasma by 96-well solid-phase extraction and liquid chromatography/tandem mass spectrometry. AB - A sensitive and rapid method based on liquid chromatography/tandem mass spectrometry (LC/MS/MS) combined with rapid solid-phase extraction (SPE) has been developed and validated for the quantitative determination of enalapril and its active metabolite enalaprilat in human plasma. After addition of internal standard to human plasma, samples were extracted by 96-well SPE cartridge. The extracts were analyzed by HPLC with the detection of the analyte in the multiple reaction monitoring (MRM) mode. This method for the simultaneous determination of enalapril and enalaprilat was accurate and reproducible, with respective limits of quantitation of 0.2 and 1.0 ng/mL in plasma. The standard calibration curves for both enalapril and enalaprilat were linear (r(2) = 0.9978 and 0.9998) over the concentration ranges 0.2-200 and 1.0-100 ng/mL in human plasma, respectively. The intra- and inter-day precision over the concentration range for enalapril and enalaprilat were lower than 13.3 and 15.4% (relative standard deviation, %RSD), and accuracy was between 89.2-105.0 and 91.9-104.7%, respectively. PMID- 12772272 TI - Approaches towards the automated interpretation and prediction of electrospray tandem mass spectra of non-peptidic combinatorial compounds. AB - Combinatorial chemistry is widely used within the pharmaceutical industry as a means of rapid identification of potential drugs. With the growth of combinatorial libraries, mass spectrometry (MS) became the key analytical technique because of its speed of analysis, sensitivity, accuracy and ability to be coupled with other analytical techniques. In the majority of cases, electrospray mass spectrometry (ES-MS) has become the default ionisation technique. However, due to the absence of fragment ions in the resulting spectra, tandem mass spectrometry (MS/MS) is required to provide structural information for the identification of an unknown analyte. This work discusses the first steps of an investigation into the fragmentation pathways taking place in electrospray tandem mass spectrometry. The ultimate goal for this project is to set general fragmentation rules for non-peptidic, pharmaceutical, combinatorial compounds. As an aid, an artificial intelligence (AI) software package is used to facilitate interpretation of the spectra. This initial study has focused on determining the fragmentation rules for some classes of compound types that fit the remit as outlined above. Based on studies carried out on several combinatorial libraries of these compounds, it was established that different classes of drug molecules follow unique fragmentation pathways. In addition to these general observations, the specific ionisation processes and the fragmentation pathways involved in the electrospray mass spectra of these systems were explored. The ultimate goal will be to incorporate our findings into the computer program and allow identification of an unknown, non-peptidic compound following insertion of its ES-MS/MS spectrum into the AI package. The work herein demonstrates the potential benefit of such an approach in addressing the issue of high-throughput, automated MS/MS data interpretation. PMID- 12772273 TI - Characterization of soluble oligomers produced by electrochemical oxidation of o phenylenediamine by electrospray ionization sequential mass spectrometry. AB - Soluble species generated during the electropolymerization of o-phenylenediamine (o-PD) on platinum electrodes in aqueous buffers at different pH values were investigated by electrospray ionization ion trap sequential mass spectrometry (ESI-ITMS(n)). The main protonated molecules (MH(+)) detected in the full scan ESI-MS spectra of the electrolytic solutions were isolated in the ion trap and sequentially fragmented (MS(n), with n up to 5) to obtain fragmentation patterns. The latter led to hypotheses as to the molecular structures of the soluble products of o-PD electropolymerization; it appeared that all of them are actually oligomeric species in different oxidation states. In particular, o-PD dimers, trimers and tetramers could be identified and three common structural features were found, namely: the presence of phenazine, 1,4-benzoquinonediimine, and secondary amine (acting as bridges between benzene rings) units. These findings are in agreement with those already reported for the surface structure of the polymeric films formed on the platinum electrodes during o-PD polymerization, thus suggesting that a close relationship exists between the soluble oligomers and the polymer itself. PMID- 12772274 TI - Silica-based monoliths for rapid peptide screening by capillary liquid chromatography hyphenated with electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry. AB - Capillary liquid chromatography based on particulate and monolithic stationary phases was used to screen complex peptide libraries by fast gradient elution coupled on-line to electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (ESI-FTICRMS). A slightly modified commercial electrospray interface consisting of a fused-silica transfer capillary and low dead volume stainless steel union at which the electrospray voltage was grounded enabled the effluent of all the capillary columns to be directly sprayed into the mass spectrometer. Stable electrospray conditions were generated over a wide range of mobile phase compositions, alleviating the need for a tapered end of the spray capillary, pneumatic assistance or preheated nebulizer gas. Since the identification of complex samples containing numerous isobaric substances is facilitated by chromatographic separation prior to mass spectrometry, stationary phase materials have been employed which offer a fast, efficient elution and, due to the complexity of samples, a high loading capacity. Silica-based monolithic capillary columns combine these three characteristics in a unique manner due to a tailored adjustment of both macro- and mesopore sizes in the highly porous silica structure. As we demonstrate by a comparative study of the silica-based monolithic and packed capillaries for LC/MS analysis of complex peptide libraries, silica monoliths show superior performance over packed beds of small diameter particles with respect to analysis time and separation efficiency. Libraries with more than 1000 different peptides could be screened in less than 20 min. PMID- 12772275 TI - Identification of dinucleoside polyphosphates by matrix-assisted laser desorption/ionisation post-source decay mass spectrometry. AB - Dinucleoside polyphosphates are a group of intra- and extracellular mediators controlling numerous physiological functions. In this study dinucleoside polyphosphates were examined by positive ion matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MADLI-TOFMS). 3 Hydroxypicolinic acid was used as UV-absorbing matrix. For the individual dinucleoside polyphosphates Ap(n)A (n = 2-7), Ap(n)G (n = 2-6) and Gp(n)G (n = 2 6), MALDI post-source decay (PSD) mass spectra were measured. Each mass peak in the MALDI-PSD mass spectra could be assigned to individual fragments of dinucleoside polyphosphates. The comparison of the fragmentation patterns of the dinucleoside polyphosphates presented here demonstrates that dinucleoside polyphosphates preferably cleave to fragment ions consisting of the corresponding mononucleoside polyphosphates as well as the corresponding nucleosides and bases during flight in the field-free drift path of the MALDI mass spectrometer. Therefore, the MALDI-PSD approach described here is suitable for identification of other dinucleoside polyphosphates. The present MALDI-PSD mass spectra may be used as MALDI-PSD mass reference spectra for future identification of dinucleoside polyphosphates and other nucleotides. PMID- 12772276 TI - A standard protocol for single nucleotide primer extension in the human genome using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. AB - Analysis of single nucleotide polymorphisms (SNPs) has become an increasingly important area of research, with numerous applications in medical genetics, population genetics, forensic science, and agricultural biotechnology. Large scale SNP analyses require the development of methodologies that are economical, flexible, accurate and capable of automation. Primer extension in conjunction with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) is currently emerging as a potential method for high-throughput SNP genotyping. We have evaluated a number of published primer extension methods and refined a simple and robust protocol to analyze human autosomal disease-causing mutations and population genetic markers on the Y-chromosome. Twelve different variant sites were examined, and homozygotes, heterozygotes and hemizygotes were accurately typed. A 100% concordance was observed between SNP genotypes obtained using the MALDI-TOFMS technique and alternative genotyping methods, such as restriction fragment length polymorphism (RFLP) assays and denaturing high performance liquid chromatography (DHPLC). Since multiple polymorphisms can be detected in single reactions, the method provides a cost-effective approach for SNP analysis. The protocol is also extremely flexible (able to accommodate new markers) and can be adapted to a number of platforms without the use of commercial kits. PMID- 12772278 TI - On-target oxidation of methionine residues using hydrogen peroxide for composition-restricted matrix-assisted laser desorption/ionisation peptide mass mapping. PMID- 12772277 TI - Observation of different ceramide species from crude cellular extracts by normal phase high-performance liquid chromatography coupled to atmospheric pressure chemical ionization mass spectrometry. AB - Normal-phase high-performance liquid chromatography (NP-HPLC) coupled to atmospheric pressure chemical ionization mass spectrometry (APCI-MS) allows qualitative analysis of endogenous ceramide and dihydroceramide species from crude lipid extracts utilizing chromatographic methods readily adaptable from commonly used thin layer chromatography (TLC) conditions. Qualitative information for the species comes from observation of differences in chromatographic and mass spectrometric behavior between species. Application to the analysis of ceramide and dihydroceramide from various cell lines is demonstrated. The results show the species profile in each cell line to be unique despite growth under identical conditions. The results from APCI-MS analysis corroborate and enhance information acquired from use of the diacylglycerol kinase assay for total ceramide measurement. This technique readily allows the previously difficult distinction between ceramide and dihydroceramide species. PMID- 12772279 TI - Investigations into the accuracy and precision obtainable on accurate mass measurements on a quadrupole orthogonal acceleration time-of-flight mass spectrometer using liquid chromatography as sample introduction. PMID- 12772280 TI - Characteristic fragmentation behavior of deoxytetranucleotides by positive ion electrospray ionization tandem mass spectrometry. PMID- 12772281 TI - Electrospray ionization mass spectrometry in the structural characterization of a mixed nitrido-Tc heterocomplex of interest for myocardial imaging. PMID- 12772283 TI - Preparation and characterization of organotin-oxomolybdate coordination polymers and their use in sulfoxidation catalysis. AB - The organotin-oxomolybdates [(R(3)Sn)(2)MoO(4)].n H(2)O (R=methyl, n-butyl, cyclohexyl, phenyl, benzyl) have been prepared and tested as catalysts for the oxidation of benzothiophene with aqueous hydrogen peroxide, at 35 degrees C and atmospheric pressure. In all cases, the 1,1-dioxide was the only observed product. The kinetic profiles depend on the nature of the tin-bound R group and also on the addition of a co-solvent. For the tribenzyltin derivative, the apparent activation energies for sulfoxidation as a function of the co-solvent are in the order 1,2-dichloroethane (5 kcal mol(-1))CH(3)F+X(-) (X=F, Cl, CN, OH, SH, NH(2) and PH(2)) have been definitively computed using focal point analyses. These analyses entailed extrapolation to the one-particle limit for the Hartree-Fock and MP2 energies using basis sets of up to aug-cc-pV5Z quality, inclusion of higher-order electron correlation [CCSD and CCSD(T)] with basis sets of aug-cc-pVTZ quality, and addition of auxiliary terms for core correlation and scalar relativistic effects. The final net activation barriers for the forward reactions are: E (b/F,F)=-0.8, E (b/F, Cl)=-12.2, E (b/F,OH)=+13.6, E b/F,OH=+16.1, E b/F,SH=+2.8, Eb/F, NH=+32.8, and E b/F,PH =+19.7 kcal x mol(-1). For the reverse reactions E b/F,F= -0.8, Eb/Cl,F =+18.3, E b/CN,F=+12.2, E b/OH,F =-1.8, E b/SH,F =+13.2, E b/NH(2),=-1.5, and E b/PH(2) =+9.6 kcal x mol(-1). The change in energetics between the CCSD(T)/aug-cc-pVTZ reference prediction and the final extrapolated focal point value is generally 0.5-1.0 kcal mol(-1). The inclusion of a tight d function in the basis sets for second-row atoms, that is, utilizing the aug-cc-pV(X+d)Z series, appears to change the relative energies by only 0.2 kcal x mol(-1). Additionally, several decomposition schemes have been utilized to partition the ion-molecule complexation energies, namely the Morokuma-Kitaura (MK), reduced variational space (RVS), and symmetry adapted perturbation theory (SAPT) techniques. The reactant complexes fall into two groups, mostly electrostatic complexes (FCH(3).F(-) and ClCH(3).F(-)), and those with substantial covalent character (NCCH(3).F(-), CH(3)OH.F(-), CH(3)SH.F(-), CH(3)NH(2).F(-) and CH(3)PH(2).F(-)). All of the product complexes are of the form FCH(3).X(-) and are primarily electrostatic. PMID- 12772293 TI - The structure of the sugar residue in glycated human serum albumin and its molecular recognition by phenylboronate. AB - Quantification of the extent of glycation of human serum albumin (HSA) and of haemoglobin provides a record of average mid- and long-term blood-sugar concentrations, respectively; this is very useful for the management of diabetes. The reaction of D-glucose with propylamine affords the corresponding Schiff base, N-propylamino-D-glucoside, in the cyclic form. This compound is not stable: upon standing or treatment with acid it is converted, by an Amadori rearrangement, into N-propylfructosamine. Both amino sugars occur predominantly in the beta pyranose form. Phenylboronate forms highly stable boronate esters through binding of the cis 1,2-diol moiety in the furanose form of N-propylfructosamine. Between pH 5 and 10, an electrostatic interaction between the protonated amino group and the negatively charged boronate moiety affords an additional stabilisation of the ester. The Schiff base, however, has no observable interaction with phenylboronate. In aqueous solution the Schiff base is in equilibrium with propylamine and glucose. Upon addition of phenylboronate, this equilibrium shifts to the side of glucose due to the formation of highly stable phenylboronate esters of the beta-furanose form of this compound. After Amadori rearrangement, the sugar moieties in glycated human serum albumin have a similar structure, they occur as an equilibrium of the beta-pyranose (59%), alpha-furanose (19%) and beta furanose (24%) anomers. The open form was not observed. The beta-furanose anomer is selectively recognised by phenylboronate. PMID- 12772294 TI - Synthesis and reactions of terminal osmium and ruthenium complexed phosphinidenes [(eta6-Ar)(L)Md=PMes*]. AB - Novel, very stable ruthenium and osmium containing terminal phosphinidene complexes [(eta(6)-Ar)(L)M=Mes*] (Ar=benzene, p-cymene; L=PR(3), CO, and RNC) have been prepared by dehydrohalogenation of novel [(eta(6)-Ar)MX(2)(PH(2)Mes*)] complexes in the presence of a stabilizing ligand. Xray crystal structures are reported for [(eta(6)-C(6)H(6))(PPh(3))Rud=PMes*] (9) and [(eta(6) pCy)(PPh(3))Os=PMes*] (4). Dehydrohalogenation in the absence of a stabilizing ligand resulted in the new P-spiroannulated Ru(2)P(2)-ring structure 16. Dehydrohalogenation in the presence of but-2-yne gave a novel phosphaallyl complex [(eta(6)-Ar)Ru(eta(3)-R(2)PC(Me)CHMe)] 26, for which an X-ray crystal structure is reported. The mechanism by which 16 and 26 are obtained is presumed to involve the intermediate formation of the 16-electron (eta(6) benzene)Rud=PMes* phosphinidene complex. PMID- 12772295 TI - Structure-nucleophilicity relationships for enamines. AB - The kinetics of the reactions of benzhydryl cations with 22 enamines, three pyrroles, and three indoles were investigated photometrically in dichloromethane. The nucleophilicity parameters N and slope parameters s of these electron-rich pi systems were derived from equation log k (20 degrees C)=s(E+N) and compared with the nucleophilicities of other pi-systems (silyl enol ethers, silyl ketene acetals) and carbanions. It is shown that the nucleophilic reactivities of enamines cover more than ten orders of magnitude, comparable to enol ethers on the low reactivity end and to carbanions on the high reactivity end. Since the products of N-attack are thermodynamically less stable than the reactants, the observed rate constants refer to the formation of the carbon bond;carbon bonds. In some cases, equilibrium constants for the formation of iminium ions were measured, which allow one to determine the intrinsic rate constants of these reactions. PMID- 12772296 TI - Low-temperature IR and NMR studies of the interaction of group 8 metal dihydrides with alcohols. AB - The reactions of the octahedral dihydrido complexes [MH(2)(PP(3))] [M=Fe, Ru, Os; PP(3)=P(CH(2)CH(2)PPh(2))(3)] with a variety of weak ROH acids have been studied by IR and NMR methods in either CH(2)Cl(2) or THF in the temperature range from 190 to 290 K. This study has allowed the determination of the spectral and thermodynamic properties associated with the formation of dihydrogen bonds (DHB) between the terminal hydrides and the OH group. Both the DHB enthalpy values and the hydride basicity factors (E(j)) have been found to increase in the order Fe < Ru < Os. The proton transfer process, leading to the DHB complexes, and eventually to eta(2)-H(2) products, has been found to depend on the acidic strength of the alcohol as well as the nature of the solvent. Low temperature IR and NMR techniques have been used to trace the complete energy profile of the proton transfer process involving the osmium complex [OsH(2)(PP(3))] with trifluoroethanol. PMID- 12772297 TI - Synthesis and characterization of ion-exchangeable titanate nanotubes. AB - Titanate nanotubes were synthesized under hydrothermal conditions. The optimized synthesis (100-180 degrees C, longer than 48 h), thermal and hydrothermal stability, ion exchangeability and consequent magnetic and optical properties of the titanate nanotubes were systematically studied in this paper. First, nanotubes with monodisperse pore-size distribution were prepared. The formation mechanism of the titanate nanotubes was also studied. Second, the thermal and hydrothermal stability were characterized with X-ray diffraction (XRD), high resolution transmission electron microscopy (HRTEM), Fourier transform infrared (FTIR), and Raman spectroscopy. Results showed that sodium ions played a significant role in the stability of the frameworks. Third, the selective ion exchangeability was demonstrated with a series of ions. The ion substitution also enlarged the BET surface area of the titanate nanotubes to 240 m(2) x g(-1). Combination of these two features implied that these nanotubes might be functionalized by substitution of different transitional-metal ions and consequently used for selective catalysis. Magnetism, photoluminescence, and UV/Vis spectra of the substituted titanate nanotubes revealed that the magnetic and optical properties of the nanotubes were modifiable. PMID- 12772298 TI - Formation of two isomeric closo-[(eta5-C5H5)FePC2B8H10] phosphadicarbaborane analogues of ferrocene via isolable eta1-bonded complexes of the [7-[(eta5 C5H5)Fe(CO)2]-(eta1-nido-PC2B8H10)] type. AB - The reaction of nido-[7,8,9-PC(2)B(8)H(11)] (1) with [[CpFe(CO)(2)](2)] (Cp=eta(5)-C(5)H(5) (-)) in benzene (reflux, 3 days) gave an eta(1)-bonded complex [7-Fp-(eta(1)-nido-7,8,9,-PC(2)B(8)H(10))] (2; Fp=CpFe(CO)(2); yield 38 %). A similar reaction at elevated temperatures (xylene, reflux 24 h) gave the isomeric complex [7-Fp-(eta(1)-nido-7,9,10-PC(2)B(8)H(10))] (3; yield 28 %) together with the fully sandwiched complexes [1-Cp-closo-1,2,4,5 FePC(2)B(8)H(10)] 4 a (yield 30%) and [1-Cp-closo-1,2,4,8-FePC(2)B(8)H(10)] 4 b (yield 5%). Compounds 2 and 3 are isolable intermediates along the full eta(5) complexation pathway of the phosphadicarbaborane cage; their heating (xylene, reflux, 24 h) leads finally to the isolation of compounds 4 a (yields 46 and 52%, respectively) and 4 b (yields 4 and 5%, respectively). Moreover, compound 3 is isolated as a side product from the heating of 2 (yield 10%). The structure of compound 4 a was determined by an X-ray structural analysis and the constitution of all compounds is consistent with the results of mass spectrometry and IR spectroscopy. Multinuclear ((1)H, (11)B, (31)P, and (13)C), two-dimensional [(11)B-(11)B]-COSY, and (1)H[(11)B(selective)] magnetic resonance measurements led to complete assignments of all resonances and are in excellent agreement with the structures proposed. PMID- 12772299 TI - Thioformaldehyde S-methylide and thioacetone S-methylide: an ab initio MO study of structure and cycloaddition reactivity. AB - The mechanisms of cycloaddition of thioformaldehyde S-methylide and thioacetone S methylide, as models for an alkyl-substituted ylide, to thioformaldehyde and thioacetone, as well as to ethene as a model for a C=C double bond have been studied by ab initio calculations. Restricted and unrestricted B3LYP/6-31G* calculations were performed for the geometries of ground states, transition structures, and intermediates. Although basis sets with more polarization functions were tested, the 6-31G* basis set was applied throughout. Single-point CASPT2 calculations are reported for analysis of the unsubstituted system. The stabilities of structures with high biradical character seem to be overestimated by DFT methods in comparison to CASPT2. The general trends of the results are independent of the level of theory. Thioformaldehyde adds to thioformaldehyde S methylide without activation energy, and the activation energies for two-step biradical pathways to 1,3-dithiolane are low. C,S biradicals are more stable than C,C biradicals. The two-step cycloaddition is not competitive with the concerted cycloaddition. Methyl substitution in the 1,3-dipole and the dipolarophile does not change the mechanistic relationships. TSs for the concerted formation of the regioisomeric cycloadducts of thioacetone Smethylide and thioacetone were located. Concerted addition remains the preferred reaction. The reactivity of the C=S double bond is high relative to that of the C=C double bond. PMID- 12772300 TI - 1,3-dithiolanes from cycloadditions of alicyclic and aliphatic thiocarbonyl ylides with thiones: regioselectivity. AB - The regiochemistry of 1,3-dithiolanes obtained from thiocarbonyl ylides 9 and thiones 10 shows a striking dependence on substituents. Previously and newly performed experiments indicate that sterically hindered cycloalkanethione S methylides and dialkylthioketone S-methylides react with alicyclic and aliphatic thiones to give the 2,2,4,4-tetrasubstituted 1,3-dithiolanes 11 exclusively. Aryl groups in one or both reactants lead to a preference for, or even complete formation of, 4,4,5,5-tetrasubstituted 1,3-dithiolanes 12. Several mechanisms appear to be involved, but the paucity of experimental criteria is troubling. Quantum-chemical calculations (see preceding paper) on the cycloaddition between thioacetone S-methylide and thioacetone furnish lower activation energies for the concerted process than for the two-step pathways via C,S- or C,C-biradicals; the favoring of the 2,4-substituted 1,3-dithiolanes over the 4,5-substituted type would be expected to increase with growing bulk of substituents. Aryl groups stabilize intermediate biradicals. Experimental criteria for the differentiation of regioisomeric dithiolanes are discussed. Thiocarbonyl ylides 9 are prepared by 1,3-cycloadditions between diazomethane and thioketones and subsequent N(2) elimination from the usually isolable 2,5-dihydro-1,3,4-thiadiazoles 17; different ratios of the two rate constants lead to divergent product formation scenarios. PMID- 12772301 TI - Novel conformationally locked inositols: from aromatics to annulated cyclitols. AB - A new family of ring-annulated inositols with "locked" conformations has been designed to deliver a range of these biologically important entities in "unnatural conformations" while retaining their "natural configurations". The simple "tool" of trans ring fusion has been used to "lock" the conformation of the annulated inositols. Short, simple syntheses of a range of these novel cyclitols have been achieved from readily available aromatic precursors such as tetralin and indane. Along the way, annulated C(2)-symmetric cyclohexadiene-trans diol (trans-CHD) derivatives have been prepared for the first time and serve as the pivotal building blocks for generating the oxy-functionalization pattern of inositols. The presence of chemo-differentiated hydroxyl groups in our novel inositols is expected to facilitate the installation of phosphate diversity to harness the biological potential of these entities. PMID- 12772302 TI - Unusual coupling reactions of aldehydes and alkynes: a novel preparation of substituted phthalic acid derivatives by automated synthesis. AB - Based upon a highly versatile multicomponent methodology, a new one-pot synthesis of substituted phthalic acid derivatives from alpha,beta-unsaturated aldehydes was developed. The reaction involves the intermediacy of an acetamidodiene species which undergoes Diels-Alder addition to diethyl acetylenedicarboxylate. The resultant acetamidocyclohexadiene is subject to elimination of acetamide under the reaction conditions to give rise to substituted diethyl phthalates in good yields. This domino condensation-cycloaddition-elimination sequence has been applied to a variety of alpha,beta-unsaturated aldehydes. Furthermore, we demonstrated the exploitation of parallelized and automated synthesis technology for the rapid screening of reaction conditions and compositions. Detailed studies revealed the catalytic role of the employed acetamide and the occurrence of a stereoselective 1,4-syn elimination pathway under standard conditions. PMID- 12772303 TI - meso-substituted aromatic 34pi core-modified octaphyrins: syntheses, characterization and anion binding properties. AB - Modified octaphyrins with 34pi electrons have been synthesized and characterized following a simple synthetic methodology. An acid-catalyzed alpha,alpha coupling of tetrapyrranes containing furan, thiophene and selenophene rings resulted in the formation of the respective octaphyrins in relatively good yield. Solution studies by (1)H NMR and 2D NMR methods and single crystal Xray structural characterization reveal an almost flat structure with two heterocyclic rings inverted. Specifically, in 14 two selenophene rings (one on each biselenophene unit) are inverted while in 15 two furan rings (one on each bifuran unit) are inverted when the meso substituent are mesityl groups. On changing the mesityl substituent to m-xylyl group as in 19, the location of ring inversion shifts to pyrrole rings (one on each bipyrrole unit) indicating the dependence of structure on the meso substituents. UV/Vis studies, both in freebase and protonated forms reveal typical porphyrinic character and the aromatic nature of the octaphyrins. The Deltadelta values evaluated by (1)H NMR spectroscopy also support their aromatic nature. The protonated forms of octaphyrins bind TFA anion in a 1:2 ratio. The TFA anions are located one above and below the plane of the octaphyrin macrocycle and they are held by weak electrostatic NH-O interactions similar to that observed for protonated rubyrins. However, in the present case, there is an additional non-electrostatic CH-O interaction involving beta-CH of the inverted heterocyclic ring and the carbonyl oxygen of the TFA. Furthermore, inter molecular interactions between the Cbond;H of the meso-mesityl group and the fluorine of CF(3) group of bound TFA leads to the formation of one-dimensional supramolecular arrays with interplanar distance of 13 A between two octaphyrins. PMID- 12772304 TI - One-step preparation, structural assignment, and X-ray study of 2,2-di-n-butyl- and 2,2-diphenyl-6-aza-1,3-dioxa-2-stannabenzocyclononen-4-ones derived from amino acids. AB - Twenty-four 2,2-di-n-butyl- and 2,2-diphenyl-6-aza-1,3-dioxa-2 stannabenzocyclononen-4-ones, each having a transannular N-->Sn bond, have been prepared by one-step reactions of alpha-amino acids (1 a-l), salicylaldehyde (2), and either di-n-butyltin(IV) oxide (3) or diphenyltin(IV) oxide (4). The new methodology constitutes an easy, highly efficient one-step synthesis of diorganotin(IV) derivatives, such as 5 a-l and 6 a-l, from iminic tridentate ligands without isolation of the Schiff bases. The structures of all the compounds have been established by a combination of (1)H, (13)C, (15)N, and (119)Sn NMR spectroscopy, IR spectroscopy, mass spectrometry, and elemental analysis. In all cases, the (119)Sn chemical shifts, as well as the (1)J((119)Sn,(13)C) coupling constants, are indicative of pentacoordinated tin atoms in solution. The structures of compounds 5 a, d, f, 6 a, b, b-racemic, c, d, f, g, and l have been established by single-crystal X-ray diffraction analyses. The tin atoms in 5 d, f, 6 a, b, b-racemic, c, d, g, and l each have a distorted trigonal-bipyramidal (TBP) geometry, with the oxygen atoms from the phenol and carboxylate moieties occupying the axial positions, and the imine nitrogen and phenyl or n-butyl substituents occupying the equatorial positions. Compounds 5 a and 6 f show distorted octahedral (DOC) geometries due to intermolecular coordination of the carbonyl oxygen to the tin atom, in a trans disposition to the N-->Sn bond, leading to trimeric 5 a and a polymeric structure for compound 6 f. Additionally, measurement of the one-bond coupling constants (1)J((119)Sn,(13)C) in diphenyltin(IV) complexes (6 a-l) and their correlation with the CSn-C bond angles has allowed the derivation of an equation that can be applied to assess the geometry around the tin atom for other diphenyltin(IV) compounds in solution. PMID- 12772305 TI - Role of the coordination of the azido bridge in the magnetic coupling of copper(II) binuclear complexes. AB - It is well-known that the azido bridge gives rise antiferromagnetic (AF) or ferromagnetic (F) coupling depending on its coordination mode, namely end-to-end or end-on, respectively. The aim of the present work is to analyse the factors contributing to this different magnetic behaviour. The difference dedicated configuration interaction (DDCI) method is applied to several binuclear Cu(II) azido-bridged models with both types of coordination. In end-on complexes, the direct exchange and the spin polarisation contributions are found to be responsible for the ferromagnetic coupling. In end-to-end complexes, both the direct exchange and the spin polarisation are small and the leading term is the antiferromagnetic dynamical polarisation contribution. The most relevant physical effects are included in the DDCI calculations so that good quantitative agreement is reached for the coupling constant as well as the spin densities. PMID- 12772306 TI - The magnetic properties of myoglobin as studied by NMR spectroscopy. AB - Deoxymyoglobin has been investigated by NMR spectroscopy to determine the magnetic anisotropy through pseudocontact shifts and the total magnetic susceptibility through Evans measurements. The magnetic anisotropy values were found to be Deltachi(ax)=-2.03+/-0.08 x 10(-32) m(3) and Deltachi(rh)=-1.02+/ 0.09 x 10(-32) m(3). The negative value of the axial susceptibility anisotropy originates from the z tensor axis lying in the heme plane, unlike all other heme systems investigated so far. This magnetic axis is almost exactly orthogonal to the axial histidine plane. The other two axes lie essentially in the histidine plane, the closest to the heme normal being tilted by about 36 degrees from it, towards pyrrole A on the side of the proximal histidine. From the comparison with cytochrome c' it clearly appears that the position of the one axis lying in the heme plane is related to the axial histidine orientation. Irrespective of the directions, the magnetic anisotropy is smaller than that of the analogous reduced cytochrome c' and of the order of that of low-spin iron(III). The magnetic anisotropy of the system permits the measurement of residual dipolar couplings, which, together with pseudocontact shifts, prove that the solution structure is very similar to that in the crystalline state. Magnetic measurements, at variance with previous data, demonstrate that there is an orbital contribution to the magnetic moment, micro(eff)=5.5 micro(B). Finally, from the magnetic anisotropy data, the hyperfine shifts of iron ligands could be separated in pseudocontact and contact components, and hints are provided to understand the spin delocalisation mechanism in S=2 systems by keeping in mind the delocalisation patterns in low-spin S=1/2 and high-spin S= 5/2 iron(III) systems. PMID- 12772307 TI - Eta6-mesityl,eta1-imidazolinylidene-carbene-ruthenium(II) complexes: catalytic activity of their allenylidene derivatives in alkene metathesis and cycloisomerisation reactions. AB - The reaction of electron-rich carbene-precursor olefins containing two imidazolinylidene moieties [(2,4,6-Me(3)C(6)H(2)CH(2))NCH(2)CH(2)N(R)Cdbond;](2) (2a: R=CH(2)CH(2)OMe, 2 b R=CH(2)Mes), bearing at least one 2,4,6-trimethylbenzyl (R=CH(2)Mes) group on the nitrogen atom, with [RuCl(2)(arene)](2) (arene=p cymene, hexamethylbenzene) selectively leads to two types of complexes. The cleavage of the chloride bridges occurs first to yield the expected (carbene) (arene)ruthenium(II) complex 3. Then a further arene displacement reaction takes place to give the chelated eta(6)-mesityl,eta(1)-carbene-ruthenium complexes 4 and 5. An analogous eta(6)-arene,eta(1)-carbene complex with a benzimidazole frame 6 was isolated from an in situ reaction between [RuCl(2)(p-cymene)](2), the corresponding benzimidazolium salt and cesium carbonate. On heating, the RuCl(2)(imidazolinylidene) (p-cymene) complex 8, with p-methoxybenzyl pendent groups attached to the N atoms, leads to intramolecular p-cymene displacement and to the chelated eta(6)-arene,eta(1)-carbene complex 9. On reaction with AgOTf and the propargylic alcohol HCtbond;CCPh(2)OH, compounds 4-6 were transformed into the corresponding ruthenium allenylidene intermediates (4-->10, 5-->11, 6-->12). The in situ generated intermediates 10-12 were found to be active and selective catalysts for ring-closing metathesis (RCM) or cycloisomerisation reactions depending on the nature of the 1,6-dienes. Two complexes [RuCl(2)[eta(1) CN(CH(2)C(6)H(2)Me(3)-2,4,6)CH(2)CH(2)N- (CH(2)CH(2)OMe)](C(6)Me(6))] 3 with a monodentate carbene ligand and [RuCl(2)[eta(1)-CN[CH(2)(eta(6)-C(6)H(2)Me(3) 2,4,6)]CH(2)CH(2)N-(CH(2)C(6)H(2)Me(3)-2,4,6)]] 5 with a chelating carbene-arene ligand were characterised by X-ray crystallography. PMID- 12772308 TI - Self-assembled monolayers of novel surface-bound dendrons: peripheral structure determines surface organization. AB - The synthetic and functional versatility of dendrimers and their well-defined shapes make them attractive molecules for surface modification. We synthesized six structurally very similar surface-bound dendrons and used them as building blocks for the preparation of self-assembled monolayers (SAMs) on a gold surface. We studied the effects of the surface-bound dendron's main structure, peripheral substituents, and the coadsorption process on its self-assembling behavior. Using scanning tunneling microscopy (STM), we observed nanostripes for SAMs of the surface-bound dendron consisting of symmetrical benzene rings. When we changed the symmetrical dendron's structure slightly, by increasing or decreasing the numbers of benzene rings at one wedge, we found no ordered structures were formed by the asymmetrical dendrons. We also introduced two kinds of substituents, heptane chains and oligo(ethylene oxide) chains, to the symmetrical dendron's periphery. Heptane chains appear to enhance the interaction between symmetrical backbones, leading to the formation of stripes, while oligo(ethylene oxide) chains appear to weaken the interaction between symmetrical backbones, resulting in a homogeneous structure. Dendrons with both heptane and oligo(ethylene oxide) chains exhibit nanophase separation in a confined state, leading to the formation of a honeycomb structure. Electrochemical studies provide additional evidence for understanding the resulting surface organizations: surface-bound dendrons with symmetrical structures form denser monolayers than their asymmetrical analogues; SAMs comprising peripherally substituted dendrons exhibit blocking effects proportionate to their hydrophilic fraction. PMID- 12772309 TI - Molecular nitrides containing group 4 and 5 metals: single and double azatitanocubanes. AB - Treatment of [[Ti(eta(5)-C(5)Me(5))(micro-NH)](3)(micro(3)-N)] (1) with the imido complexes [Ti(NAr)Cl(2)(py)(3)] (Ar=2,4,6-C(6)H(2)Me(3)) and [Ti(NtBu)Cl(2)(py)(3)] in toluene affords the single azatitanocubanes [[Cl(2)(ArN)Ti]( micro(3)-NH)(3)[Ti(3)(eta(5)-C(5)Me(5))(3)(micro(3) N)]].(C(7)H(8)) (2.C(7)H(8)) and [[Cl(2)Ti](micro(3)-N)(2)(micro(3) NH)[Ti(3)(eta(5)-C(5)Me(5))(3)(micro(3)-N)]] (3), respectively. Similar reactions of complex 1 with the niobium and tantalum imido derivatives [[M(NtBu)(NHtBu)Cl(2)(NH(2)tBu)](2)] (M=Nb, Ta) in toluene give the single azaheterometallocubanes [[Cl(2)(tBuN)M](micro(3)-N)(micro(3)-NH)(2)[Ti(3)(eta(5) C(5)Me(5))(3)(micro(3)-N)]] (M=Nb (4), Ta (5)), both complexes react with 2,4,6 trimethylaniline to yield the analogous species [[Cl(2)(ArN)M](micro(3) N)(micro(3)-NH)(2)[Ti(3)(eta(5)-C(5)Me(5))(3)(micro(3)-N)]].(C(7)H(8)) (Ar=2,4,6 C(6)H(2)Me(3), M=Nb (6.C(7)H(8)), Ta (7.C(7)H(8))). Also the azaheterodicubanes [M[micro(3)-N)(2)(micro(3)-NH)](2)[Ti(3)(eta(5)-C(5)Me(5))(3)(micro(3) N)](2)].2C(7)H(8) [M=Ti (8.2C(7)H(8)), Zr (9.2C(7)H(8))], and [M[(micro(3) N)(5)(micro(3)-NH)][Ti(3)(eta(5)-C(5)Me(5))(3)(micro(3)-N)](2)].2 C(7)H(8) (Nb (10.2C(7)H(8)), Ta (11.2C(7)H(8))) were prepared from 1 and the homoleptic dimethylamido complex [M(NMe(2))(x)] (x=4, M=Ti, Zr; x=5, M=Nb, Ta) in toluene at 150 degrees C. X-ray crystal structure determinations were performed for 6 and 10, which revealed a cube- and double-cube-type core, respectively. For complexes 2 and 4-7 we observed and studied by DNMR a rotation or trigonal-twist of the organometallic ligands [[Ti(eta(5)-C(5)Me(5))(micro-NH)](3)(micro(3)-N)] (1) and [(micro(3)-N)(micro(3)-NH)(2)[Ti(3)(eta(5)-C(5)Me(5))(3)(micro(3)-N)]](1-). Density functional theory calculations were carried out on model complexes of 2, 3, and 8 to establish and understand their structures. PMID- 12772310 TI - A non-radical mechanism for methane hydroxylation at the diiron active site of soluble methane monooxygenase. AB - We propose a non-radical mechanism for the conversion of methane into methanol by soluble methane monooxygenase (sMMO), the active site of which involves a diiron active center. We assume the active site of the MMOH(Q) intermediate, exhibiting direct reactivity with the methane substrate, to be a bis(mu-oxo)diiron(IV) complex in which one of the iron atoms is coordinatively unsaturated (five coordinate). Is it reasonable for such a diiron complex to be formed in the catalytic reaction of sMMO? The answer to this important question is positive from the viewpoint of energetics in density functional theory (DFT) calculations. Our model thus has a vacant coordination site for substrate methane. If MMOH(Q) involves a coordinatively unsaturated iron atom at the active center, methane is effectively converted into methanol in the broken-symmetry singlet state by a non radical mechanism; in the first step a methane C-H bond is dissociated via a four centered transition state (TS1) resulting in an important intermediate involving a hydroxo ligand and a methyl ligand, and in the second step the binding of the methyl ligand and the hydroxo ligand through a three-centered transition state (TS2) results in the formation of a methanol complex. This mechanism is essentially identical to that of the methane-methanol conversion by the bare FeO(+) complex and relevant transition metal-oxo complexes in the gas phase. Neither radical species nor ionic species are involved in this mechanism. We look in detail at kinetic isotope effects (KIEs) for H atom abstraction from methane on the basis of transition state theory with Wigner tunneling corrections. PMID- 12772311 TI - Direct catalytic asymmetric mannich reactions of malonates and beta-keto esters. AB - The first catalytic asymmetric direct Mannich reaction of malonates and beta-keto esters has been developed. Malonates react with an activated N-tosyl-alpha-imino ester catalyzed by chiral tert-butyl-bisoxazoline/Cu(OTf)(2) to give the Mannich adducts in high yields and with up to 96% ee. These reactions create a chiral quaternary carbon center and it is demonstrated that this new direct Mannich reactions provides for example a new synthetic procedure for the formation of optically active beta-carboxylic ester alpha-amino acid derivatives. A series of different beta-keto esters with various ester substituents has been screened as substrates for the catalytic asymmetric direct Mannich reaction and it was found that the best results in terms of yield, diastereo- and enantioselectivity were obtained when tert-butyl esters of beta-keto esters were used as the substrate. The reaction of different beta-keto tert-butyl esters with the N-tosyl-alpha imino ester gave the Mannich adducts in high yields, diastereo- and enantioselectivities (up to 95% ee) in the presence of chiral tert-butyl bisoxazoline/Cu(OTf)(2) as the catalyst. To expand the synthetic utility of this direct Mannich reaction a diastereoselective decarboxylation reaction was developed for the Mannich adducts leading to a new synthetic approach to attractive optically active beta-keto alpha-amino acid derivatives. Based on the stereochemical outcome of the reactions, various approaches of the N-tosyl-alpha imino ester to the chiral bisoxazoline/Cu(II)-substrate intermediate are discussed. PMID- 12772312 TI - A new strategy for the design of water-soluble synthetic receptors: specific recognition of DNA intercalators and diamines. AB - Water-soluble zinc bisporphyrin receptors 1 and 2 having two Lewis acidic sites (zinc) in the hydrophobic environment consisting of alkyl chains and a bisporphyrin framework, and covered with hydrophilic exterior (twelve or eighteen carboxyl groups) were prepared. The receptors show high affinity for diamines and DNA intercalators in water where the binding constants K(a) are of the order of 10(7) and 10(8) M(-1), respectively. Diamines and DNA intercalators are bound to the receptor through different mechanisms. Diamines are bound through hydrophobic interactions and zinc-nitrogen interactions, while DNA intercalators are bound through hydrophobic interactions and charge-transfer interactions. Flexible alkyl chains can make van der Waals contact with guests and create a hydrophobic environment around the bound guest by an induced-fit-type mechanism. For the binding of DNA intercalators, the following features are noteworthy: 1). Binding constants are similar between the zinc porphyrins and zinc-free porphyrins; 2). the binding constant is larger for the guest having the lower LUMO; this indicates the important contribution of charge-transfer interactions to binding; 3). the hydrophobic and cationic nature of DNA intercalators is substantially important, and 4). higher ionic strength reduced the binding affinities; this shows a moderate contribution of electrostatic interactions. The conformational instability of the receptors also contributes to the tight binding: hydrophobic and electrostatic interactions cannot both be favorable at the same time in the guest-free receptor. Enthalpy-entropy compensation observed for the binding of diamines and DNA intercalators is characterized by a relatively small slope (alpha=0.74) and a large intercept (beta=7.75 kcal mol(-1)) in the DeltaH degrees versus TDeltaS degrees plot; this shows that a conformational change of receptors and a significant desolvation occur upon binding. The receptor can competitively bind to propidium iodide to deprive DNA of the intercalated propidium iodide. These features of water-soluble receptors consisting of a rigid framework and flexible side chains with a large solvent-accessible area are in contrast to highly preorganized rigid receptors, and they can provide useful guidelines for rational design of induced-fit artificial receptors in water. PMID- 12772314 TI - [Gene chip screen in allergic rhinitis and preliminary analysis of the differential expressed genes]. AB - OBJECTIVE: To discover the etiology of allergic rhinitis(AR) at the genomic level. METHODS: Microarray analysis on a genomic scale was used to find changes in genes expression in AR. The total RNAs were isolated from nasal mucosa of AR and normal, then purified to mRNAs and were reversely transcribed to cDNAs with the incorporation of fluorescent-labeled dUTP to prepare for the hybridization probes. The genes differential expressed in AR were screened with the gene chip which contained 12,800 human genes in all 8 samples. RESULT: 734 genes were shown in differential expressed profile with 430 upregulated genes and 304 downregulated genes, and all 8 samples were 15 genes upregulated and 6 genes downregulated. The differential expressed genes involved in the functions of inflammation, immunology, neuroendocrine and signal conduct, and they had potential value in AR study. CONCLUSION: The investigation based on gene chip in researching related genes in AR afforded a new idea in studying pathogenesis of nasal allergic diseases. PMID- 12772315 TI - [Allergic fungal sinusitis: report of three cases]. AB - OBJECTIVE: To discuss and analyze the diagnosis and management of allergic fungal sinusitis. METHOD: Three middle-aged patients with allergic fungal sinusitis (AFS) were observed. The histological diagnosis was made on the mucous material and allergy evaluation based on the strong history of inhalant mold allergies, a positive result of skin test, total immunoglobulin E level and radioallergosorbent test to fungal antigens. The management included wide local endoscopic sinus debridement, adequate sinus aeration, postoperative use of systemic and intranasal corticosteroid. Systemic antifungal therapy was not used. RESULTS: All 3 patients were immunocompetent and none demonstrated histologic evidence of tissue invasion. Two patients remained disease-free during follow-up ranging for a mean of 16 months, 1 patient recurred 17 months after operation and symptom-free for 2 months after re-cleaning of the sinus cavity. CONCLUSION: Diagnosis of AFS required a high index of suspicion. A confirmatory diagnosis was made from the inspissated mucus, clinical and CT findings along with careful communication with mycologists and immunologists about the possibilities of fungal growth and allergy. Costly and lengthy courses of antibiotics, aggressive surgery should be avoided. Surgical debridement and systemic corticosteroids followed by intranasal corticosteroids might provide long-term control of AFS. PMID- 12772316 TI - [Expression of tenascin and fibronectin in nasal polyps]. AB - OBJECTIVE: To explore the role of tenascin (TN) and fibronectin (FN) in the pathophysiology of nasal polyp. METHODS: The expression of TN and FN in nasal polyps from 34 patients and in inferior turbinates from 20 patients with deviation of nasal septum was studied with immunohistochemical method. In patients with nasal polyps, the relations between expression and histopathologic characteristics, eosinophilias (EOS) infiltration, clinical staging and the size of nasal polyps were analyzed. RESULTS: (1) The gray score of TN and FN expression was 163.10 +/- 10.54 and 163.24 +/- 11.52 in nasal polyps respectively, whereas it was 175.49 +/- 9.29 and 173.93 +/- 7.92 in inferior turbinates respectively. The difference between two groups was significant(P < 0.01); (2) The expression of TN and FN in edematous type was significantly stronger than that in cystic fibrous and glandular type (P < 0.05); (3) The association between FN expression and EOS infiltration was significant(r = -0.60, P < 0.01); (4) The expression of TN and FN did not correlate with clinical staging and size(P > 0.05). CONCLUSION: Abnormal ECM may contribute to proliferation of epithelium, accumulation of EOS and edema formation, as a result, to enhance the development of nasal polyps. PMID- 12772317 TI - [Measurement of nasal airway resistance and olfactory function before and after endoscopic sinus surgery]. AB - OBJECTIVE: To quantitatively analyse the changes of nasal airway resistance (NAR) and olfactory function in patients before and after endoscopic sinus surgery(ESS). METHODS: NAR and olfactory functions in 127 patients suffering from chronic sinusitis and/or nasal polyps were measured with anterior rhinomanometry and T&T olfactometer standard odors. RESULTS: 1. The degree of NAR increased and olfactory dysfunction were accompanied with varied clinic classifications. 2. After the ESS, NAR decreased and olfactory functions improved obviously, the effective rates were 93.4% (85/91) and 71.9% (64/89) respectively. 3. Allergy was one of the major factors that affected the outcome of nasal function after ESS. CONCLUSIONS: 1. The improvements of NAR and olfactory function in patients with chronic sinusitis and polyps were significant after ESS. 2. As the determining method of nasal function, the measurement of NAR and olfactory function ought to be used widely in clinical practice. PMID- 12772318 TI - [Effects of botulinum toxin-A on facial motoneurons]. AB - OBJECTIVE: To examine effects of botulinum toxin-A(BTX-A) intramuscular injection at rat whisker pad on facial motoneurons. METHODS: A total of 54 male Sprague Dawley rats were used in the present study. A single intramuscular injection of the BTX-A at rat whisker pad as model was employed, using retrograde labeling and immunohistochemical staining methods, to observe the effects of BTX-A on the facial motoneurons survival and the expression of choline acetyltransferase (ChAT) and vescular acetylcholine transporter (VAChT). Rats received intramuscular injection of 0.9% NaCl served as control. RESULTS: The fluorescence labeled facial motoneurons were all in the lateral subnucleus of facial nucleus after BTX-A injection at rat whisker pad. There was no significant difference of labeled motoneuron number in BTX-A injection group vs control at observed time points (t = 0.022. P > 0.05). At 7 d after BTX-A injection, optical density of ChAT and VAChT immunostaining began to increase in facial motoneurons of the lateral subnucleus ipsilateral to the injection side. A maximal increase of this data was observed 14 d after injection; thereafter it declined gradually to normal level. The patterns of changes in optical density of ChAT and VAChT immunostaining in facial motoneurons of the lateral subnucleus contralateral to the injection side were similar to that of ipsilateral. There were significant difference of optical density of ChAT and VAChT immunostaining in facial motoneurons of the lateral subnucleus in BTX-A injection group vs control(t = 3.054, P < 0.05; t = 3.366, P < 0.05). CONCLUSION: After BTX-A injection at rat one side whisker pad, the number of facial motoneurons innervating muscles of whisker pad in the lateral subnucleus has no change. However, the expression of ChAT and VAChT increase in the facial motoneurons of the lateral subnucleus. These data indicate that BTX-A intramuscular injection at facial nerve target organ doesn't induce the loss of facial motoneurons, and only results in the expression of protein related to acetylcholine changing. PMID- 12772319 TI - [Measurement of facial nerve blood flow in rabbits using laser Doppler flowmetry]. AB - OBJECTIVE: To study the microcirculation disorder of the facial nerve entrapment in rabbits. METHODS: Laser Doppler Flowmetry (LDF) was used in the blood flow measurement of the horizontal segment of the facial nerve and other adjacent structures in 32 New Zealand White rabbits. The repeated test was used to evaluate the stability of LDF. RESULTS: An animal model was established to study the microcirculation of the facial nerve in rabbits. The record of repeat test showed that LDF is a stable method. LDF of the horizontal segment of the facial nerve is higher in the proximal end but lower in distal. LDF in the distal part of the horizontal facial nerve (57.96 +/- 24.59) is higher than the semicircle canal(21.16 +/- 6.39), the medial wall of the attic(21.17 +/- 6.21), the tendon of the tensor tympani muscle(10.40 +/- 5.97) and other adjacent structures (P < 0.05). LDF probe may become a detector for surgeon to find the facial nerve in the complicated operation fields and prevent the iatrogenic facial paralysis. Because the press on the nerve can induce the decrease of the LDF, we estimate LDF may be very useful in the study of facial nerve entrapment. CONCLUSIONS: LDF is a new, stable, objective method for microcirculation study on facial nerve, and very useful in pathophysiological investigation of peripheral facial paralysis. PMID- 12772320 TI - [Enzyme histochemistry study of non-focused ultrasound irradiating the cochlea in guinea pigs]. AB - OBJECTIVE: To study the enzyme histochemistry changes of non-focused ultrasound (NFU) irradiating the cochlea in guinea pigs. METHODS: After 30 minutes or 8 hours of 2.5 MHz, 8 MHz NFU irradiating the cochlea in guinea pigs (70 ears) for 6 hours, the SDH activity of the hair cells of the cochlea was observed. RESULTS: The non-focused ultrasound (2.5 MHz or 8 MHz) irradiation for 6 hours has lad to enervation of the activity of SDH in hair cells, especially in outer hair cells, in the 0.87 +/- 0.20 mm areas of the second turn or in the 0.80 +/- 0.20 mm areas of the first turn, respectively. The activity of SDH can be part restorable in the 8 hours groups. CONCLUSION: NFU (2.5 MHz, 8 MHz) irradiating the cochlea for 6 hours can cause pathological changes in the hair cells of in different areas of cochlear. Moreover, the pathological changes could be restorable or part restorable in some amounts of NFU irradiation. The results suggest that the cochlear hair cell of the different area might be related to ultrasonic perception. PMID- 12772321 TI - [Effects of mtDNA deletion associated with abnormal expression in rat cochlear with presbycusis]. AB - OBJECTIVE: To investigate the effects of the common deletion (4834-bp) of mitochondrial DNA (mtDNA) in rat cochlear with presbycusis. METHODS: The mtDNA 4834-bp deletion was analyzed by PCR. The mitochondrial-encoded cytochrome c oxidase subunit I (COXI) transcript level and cytochrome c oxidase (COX) activity were measured by RT-PCR and histochemical methods respectively. RESULTS: The 4834 bp deletion occurred in all the senescent rat cochlear. The COXI transcript level was decreased associated with the decline of COX activity. CONCLUSION: The mtDNA 4834-bp deletion presented in the rat cochlear with presbycusis, which lead to the decrease of COXI transcript level and COX activity, may play an important role in the pathogens of presbycusis. PMID- 12772322 TI - [Effects of alcoholism on auditory event-related potential in fetal rats]. AB - OBJECTIVE: To study the effects of embryonic alcoholism, with an animal model of auditory event-related potential (AERP) recorded in the conditions of various alcohol dosages administrated during the auditory center nervous system development, on the perceptive functions in young Sprague-Dawley rats. METHODS: Normal female rats were divided into 3 groups. From the days 8.5 to 14 after mating, pregnant rats were fed 75% analytical reagent alcohol via stomach tube with 2.0, 1.0 and 0.1 g.kg-1.d-1 respectively. On the days 30-40 after birth, AERP was recorded in young rats with passive mode under anesthesia. Normal young rats with parallel age were chosen as the normal group. RESULTS: AERP was not affected after administration of the low dosage alcohol. Nevertheless, AERP was affected after administration of the middle and high dosages, manifesting the incidence rate lowing or response disappearing, latency delaying, amplitude reducing and waveform distorting, especially shorter waves of AERP. CONCLUSION: The alcoholism would produce apparent effects with dosage-effect relationship on the responsibility of auditory center during development in fetal rats, which reflected that there was a direct tie between the responsibility of AERP and the function of auditory center. PMID- 12772323 TI - [Missed diagnostic cholesteattomaous otitismedia]. AB - OBJECTIVE: To explore the missed diagnostic cholesteattomaous otitismedia lacked of clinical symptoms in order to minimize the mistake in clinical diagnose. METHODS: A retrospective study of 31 cases with the missed diagnostic cholesteattomaous otitismedia, confirmed by surgery and pathology, was conducted. RESULTS: Twenty-one cases (67.7%) had not obvious otorrhea. There were only slight finding such as attic retraction, apophysis, small granulation tissue, ear wax covered or tiny, even non, perforation in the pars flaccida or the superior and posterior pars tensa of the ear-drum. In general, the hearing loss is slight or medium conductive deafness. The type B curve was showed in tympanogram. The diagnosis rate of X-ray was only 41.9%, and 80.7% for the computed tomography. The middle ear structures aggressed by cholesteames was epitympanum, ossicular chain, tympanic antrum, mastoid process and mesotympanum. The complications occurred in 11 cases (35.5%) were the expose of facial nerve or and the meninges, or, and the labyrinth fistula. CONCLUSION: The cases with the cholesteatoma lack obvious clinic symptoms would result in the missed diagnosis because of slow and long term during cholesteatoma development. Sometimes misdiagnosed or missed diagnosis happened to in such cases of cholesteatoma with little clinical manifestation, and it is so dangerous for the patients in such case. Therefore, otolaryngologist must pay more attention to the missed diagnostic cholesteattomaous otitismedia. PMID- 12772324 TI - [Management of sinonasal inverting papilloma with recurrence and malignant transformation]. AB - OBJECTIVE: To investigate the clinical characteristics and optimum management of sinonasal inverting papilloma with recurrence and malignant transformation. METHODS: Retrospective analysis of inverting papilloma with recurrence and malignant transformation treated from 1990 to 2000 in PLA general hospital. RESULTS: All the 22 patients with recurrence had the history of sinonasal surgery. Among them, 13 patients recurred once, 7 recurred 2 times, 1 recurred 3 times and 1 recurred 4 times. The recurrence time was from 3 months to 11 years. Among the 22 patients with recurrence, the recurrence ratio of different surgery was different, for example intranasal approach was 100%, maxillectomy 60% and lateral rhinotomy 42%, 10 patients were operated with endoscope or endoscopic assisted surgery with no recurrence after 1-5 year's follow-up. Among 8 patients with malignant transformation, 4 received lateral rhinotomy, 1 Caldwell-Luc operation, all of them were given radiation therapy after operation (40-70 Gy) and no recurrence were found during 2 to 6 years follow-up. Of the other 3 patients, 2 died and 1 lost follow-up. CONCLUSION: The endoscopic surgery(including the method of endoscopy combined with extranasal incision) is the optimum management for inverting papilloma in nasal cavity and paranasal sinuses. To the patients with malignant tumors, extensive excision and post operative radiotherapy should be considered, but the prognosis is not satisfactory. PMID- 12772325 TI - [Traumatic optic neuropathy: importance and result of surgical treatment]. AB - OBJECTIVE: To explore the importance and result of surgical treatment in traumatic optic neuropathy (TON). METHODS: The results of optic canal and orbital apex decompression by the endonasal endoscopic approach were analyzed in 52 cases (53 eyes), follow up 6 months to 3 years. RESULTS: Twenty one eyes (39.6%) consisting of 14 eyes without effect by corticosteroids treatment(14/37, 37.8%) and 7 eyes with single surgery(7/16, 43.8%) showed improved vision after surgery. 18 eyes with no light perception (18/49, 36.7%) and 3 eyes with poor sight (3/4, 75.0%) had improved vision after surgery. CONCLUSION: The surgical treatment is important for elevating curative effect of TON. PMID- 12772326 TI - [Reinnervation of the posterior cricoarytenoid muscle by the phrenic nerve for bilateral vocal cord paralysis in humans]. AB - OBJECTIVE: To reestablish the respiratory abduction of the paralyzed vocal cord through reinnervation of the posterior cricoarytenoid(PCA) muscle by the phrenic nerve in humans. METHODS: In six cases with bilateral recurrent laryngeal nerve paralysis, the phrenic nerve was anastomosed to the anterior branch of recurrent laryngeal nerve, while the adductor branch of recurrent laryngeal nerve was severed and its proximal end was implanted into the PCA muscle belly in one side, for the other side nerve-muscle pedicle technique was used. All cases had been subjected to preoperative and postoperative video laryngoscopy, stroboscopy, electromyography, voice recording and acoustic analysis. RESULTS: Among the 6 patients, it is observed in five cases' phrenic nerve reinnervation side the inspiratory abducent motion evidently recovered, and the abducent range was from 3 to 5 mm, While only slight abductent motion or no motion could be recorded on the other side reinnervated with nerve-muscle pedicle technique, and the vocal cord excursion on this side was less than 1 mm in all cases. It is because the glottis is broad enough for the patients to have daily activities without short of breath, so all of them were decannulated postoperatively. The reinnervated PCA muscle by the phrenic nerve showed typical inspiratory high frequency discharge with 100-200 ms delay as compared with the other side, indicating the phrenic motoneuron pattern. No long-term diaphragmatic paralysis and lesion of respiratory function was found. All cases' voice was not weakened, and no aspiration occurred. CONCLUSION: The phrenic reinnervation is feasible clinically for treating vocal cord paralysis, and it is found to be more effective for restoring inspiratory abducent function than the nerve-muscle pedicle technique. PMID- 12772327 TI - [Research into the human leucocyte antigen I deletion and local lymphocytes infiltration in laryngeal carcinoma]. AB - OBJECTIVE: To detect the deletion of human leucotyte antigen (HLA) class I antigen expression in laryngeal carcinoma tissue and discuss the relationship between its expression and tumor infiltrating lymphocyte (TIL) cell infiltration or/and malignancy of tumor cell. METHODS: From Feb 2001 to May 2001 specific mouse antihuman HLA class I monoclonal antibody was combined to examined tissue, 64 samples of laryngeal carcinoma were detected by streptavidin-peroxidase (SP) method immunohistochemistry. RESULTS: All of 64 cases, 57 were positive HLA class I antigen and 7 were negative, the negative expression rate was 10.9% (7/64). CD3+ and CD8+ T cell infiltrated in HLA class I positive tumor mass were significantely more than those in negative tissue. the malignant degree elevated along with the declination of HLA class I antigen expression. By 3 years follow up, the mortalitywas not statistically significant in both HLA- group and HLA+ group (P > 0.05). CONCLUSION: There is deletion of HLA class I antigen in laryngeal tissue, which favours penetration of cytotoxic lymphoid cells into tumor mass; alteration in HLA class I expression may be used by cancer cells to avoid immune surveillance. PMID- 12772328 TI - [Evaluate the curative effect of CO2 laser in treatment of glottic carcinoma]. AB - OBJECTIVE: To evaluate the curative effect of CO2 laser in treatment of glottic carcinoma. METHODS: Retrospective analysis of 217 cases of early glottic laryngeal carcinoma treated with laser surgery were carried out. Among these cases, Tis were 22 cases, T1a 108 cases, T1b 38 cases, T2 46 cases, T3 3 cases. 1 case being applied vocal cord excision due to failure of tumor exposure. The follow-up period was 3 to 9 years. RESULTS: Relapse were detected in 21 out of 217 cases of glottic laryngeal carcinoma after CO2 laser surgery under self retaining laryngoscope. The recurrent rate was 9.7% (21/217). Recurrent ratio of T1a, T1b, T2, T3 were 5.6% (6/108), 21% (8/38), 13.0% (6/46), 1/3 respectively, with significant differences among groups (chi 2 = 6.102, P < 0.01). The recurrent rate was 21.6% of tumors offended the anterior commissure, versus 5.1%(8/157) with no involvement of anterior commissure (chi 2 = 13.64, P < 0.01). 217 cases received laser surgery as the only treatment, 201 cases were still alive. 4 failed to be followed-up(taken into dead number), 12 cases were dead. 3 year survival rate was 97.2%(211/217), 5 year survival rate was 89.4%(118/132). CONCLUSION: It was reliable to use laser surgery in treatment of early glottic laryngeal cancer. The advantages of it included lower complication rate and vocal function was well retained. PMID- 12772330 TI - The state Medicaid crisis: benefits and eligibility on the block. PMID- 12772329 TI - [Utility of spiral computed tomography in the study of dislocation of cricoarytenoid joint]. AB - OBJECTIVE: To demonstrate the cricoarytenoid relationship presented with spiral computed tomography and to show that the reconstraction of arytenoid dislocation can be presented by using multiplanar reconstruction algorithms. METHODS: Fifteen patients with arytenoid dislocation documented by fiberoptic laryngoscopy and strobovideolaryngoscopy and 10 normal person were displayed with spiral CT. A making design of our own has been used to diagnose arytenoid dislocation on axial CT image. RESULTS: Dislocation of cricoarytenoid joint were consistently demonstrated on several of the overlapping thin axial reconstructions in each of the 15 patients, in whom asymmetry of the bilateral cricoarytenoid joints was noted on axial images. Especially, by means of ordinary protractor, it has been found on the glottic-fissure level that the basal angle of abnormal side in 8 patients is larger than the one of normal side, and smaller in 7 patients, the value-difference of two basal angles ranges from 5 to 45 in 15 patients, 13 of whom ranges from 10 to 30. Whereas right basal angle is equal to left one in 8 except 2 whose value-difference of two basal angles is very small, respectively 2 and 3, in 10 normal person. High-quality sagittal and coronal reconstructive images often were helpful in confirming or clarifying the complex arytenoid orientations. CONCLUSION: The findings that two-side basal angle is not equal in triangle of glottic fissure can be used as a parameter to arytenoid dislocation. Spiral CT is a useful adjunct in diagnosing and treating dislocation of cricoarytenoid joint. PMID- 12772331 TI - Lessons from Canada about a single-payer system. PMID- 12772332 TI - James Peppiatt-Combes, Executive Vice President and CFO, Trinity Health, Novi, Michigan. PMID- 12772333 TI - Closing the gap in market share. PMID- 12772334 TI - Selling real estate to meet capital needs. AB - Real estate can provide a means for hospitals to raise capital. Selling a building and investing proceeds in revenue-producing operations may yield greater return than rental income. In a ground lease, a hospital can require the buyer to adhere to certain limitations that are beneficial to the hospital's strategic goals. PMID- 12772335 TI - Surefire strategies to reduce claim denials. AB - Denials can be reduced by undertaking some simple steps: Transfer patients automatically from scheduling to pre-registration to have benefits verified and authorizations obtained. Make sure system data fields are large enough to enter all information needed. Track the frequency of denials for noncontracted services and either include them in the contract at renegotiation or discontinue the billing. PMID- 12772336 TI - The capital challenge needs versus resources. PMID- 12772337 TI - The risks and rewards of swaps. PMID- 12772338 TI - Today's key drivers of physician loyalty. PMID- 12772339 TI - Developing an assessment process for new technologies. PMID- 12772340 TI - Core competencies: a simplified look at a complicated issue. PMID- 12772341 TI - HIPAA compliance: have you completed a risk analysis? PMID- 12772342 TI - Synthetic refunding: a financial tool for a low interest-rate environment. PMID- 12772343 TI - A great honor; great responsibility. PMID- 12772344 TI - 100 percent! Care model increases patient and employee satisfaction. PMID- 12772345 TI - EI, not IQ, is the key to outstanding leadership. PMID- 12772346 TI - How does excellence become an everyday occurrence? PMID- 12772347 TI - Connecting for health. PMID- 12772348 TI - Developing new and current employees. PMID- 12772349 TI - Hospitalists increase care. PMID- 12772350 TI - Exploring some of Michigan's best practices. PMID- 12772351 TI - "IT" outsourcing gives Michigan hospitals a boost. PMID- 12772352 TI - Healthy hearts. Technology that detects possible heart disease, cancer and more. PMID- 12772353 TI - How to keep your customers. PMID- 12772354 TI - Do you influence? PMID- 12772355 TI - Good news about hospitals. PMID- 12772356 TI - Whiplash: still a pain in the neck. PMID- 12772357 TI - Whiplash: still a pain in the neck. PMID- 12772358 TI - The sexually transmitted infection 'check up'. AB - BACKGROUND: Many sexually transmitted infections (STIs) and blood borne viruses (BBV) such as HIV are asymptomatic. Early detection is important for minimising associated risks. With appropriate treatment and management (including contact tracing) it is possible to substantially reduce morbidity as well as transmission to sexual partners and the neonate. OBJECTIVE: This paper outlines which tests should be administered to otherwise 'well' individuals. It also examines the questions of when, why and how to respond to requests for an STI 'screen' or 'check up'. DISCUSSION: Testing and screening for asymptomatic STIs and BBV are important, especially in situations where proven interventions can decrease morbidity and transmission. Screening for STIs also provides the opportunity in a one-on-one consultation for health promotion. Sexually transmitted infection testing can also initiate a conversation about 'safer sex' and may help address other concerns patients may have. PMID- 12772359 TI - Pelvic inflammatory disease. AB - BACKGROUND: Pelvic inflammatory disease (PID) is a treatable condition with serious long term sequelae. Due to the wide spectrum of disease and clinical presentation, diagnosis can often be challenging. OBJECTIVE: This article discusses the diagnosis, management and treatment of PID in the general practice setting. DISCUSSION: Prompt and effective treatment is essential in cases of PID. A high index of suspicion for symptomatic women at risk may help decrease the burden of serious associated morbidity. PMID- 12772360 TI - Human papillomavirus: beware the infection you can't see. AB - BACKGROUND: Genital human papillomavirus (HPV) is a common sexually transmitted infection. In the first 10 years of sexual activity, point prevalence rates approach 25% and the lifetime risk of acquisition of this infection may be as high as 80%. There are over 200 types of HPV, of which approximately 50 infect the genital area. OBJECTIVE: This article aims to discuss HPV detection and its role in cervical cancer development. DISCUSSION: The HPV types that cause genital warts do not cause cervical cancer. The subclinical types (especially types 16 and 18) are most frequently found in high grade epithelial abnormalities and therefore can potentially cause anogenital cancers. Human papillomavirus is a 'necessary but not sufficient cause for cervical cancer'. Most genital HPV infection is transient. Cervical cancer is actually a rare outcome of HPV infection. However, only 5% of women in developing countries have had a Pap smear in the past five years, and worldwide approximately 250 thousand women die of this disease every year. The role of HPV DNA testing has not yet been defined, but is no doubt a potential tool for the future. Meanwhile, international vaccine trials using HPV virus-like particles are taking place, and look promising. PMID- 12772361 TI - Recognising acute HIV infection. AB - BACKGROUND: Early diagnosis and appropriate management of acute or primary human immunodeficiency virus (HIV) infection may significantly alter the long term course of the disease and help reduce further transmission. OBJECTIVE: General practitioners are the doctors most likely to see a person with acute or primary HIV infection. This article aims to assist GPs in the early diagnosis and management of these patients. DISCUSSION: The symptoms of acute HIV infection include headache, fever, malaise and rash. Good history taking including assessing possible HIV transmission risk and careful examination can alert GPs to the illness. Appropriate investigations require knowledge of the time course of serological and virological markers and expert laboratory advice may be needed. Management may include general supportive measures for physical, psychological and social issues and some clinicians advocate the use of antiretrovirals. Postexposure prophylaxis is becoming more widely available in Australia. Most patients can be cared for by their own GP with the support and advice of a more HIV experienced colleague. PMID- 12772363 TI - Patient education. How to use a condom. PMID- 12772362 TI - Patient education. What is safer sex? PMID- 12772365 TI - Primary health care for adults with intellectual disability. AB - BACKGROUND: In recent decades people with intellectual disability have moved from institutional care into the community and are now reliant on general practice services for their primary medical care. Their complex health needs present a considerable challenge to the general practitioner. OBJECTIVE: This article aims to outline the common physical and psychiatric conditions found in adults with intellectual disability, and the need for a structured approach to health care in patients with limited communication skills. DISCUSSION: Adults with intellectual disability frequently have undiagnosed health problems. In many cases these will only be identified by specific examination and testing. Health assessment tools offer a useful guide to which conditions to look out for. Early identification and treatment enhances quality of life for both people with intellectual disability and their carers. PMID- 12772364 TI - Which drug class and why? AB - BACKGROUND: Having made the diagnosis, decided on therapeutic goals and the therapeutic approach, the next step is choice of drug class. OBJECTIVE: This article discusses the four factors to be considered when choosing an appropriate drug class: efficacy, safety, suitability and cost. DISCUSSION: The first consideration in choosing a drug class is the relative efficacy compared with other agents for the particular therapeutic goal. Safety is a broad issue involving adverse reactions, toxicity, tolerance and dependence, teratogenicity, and consideration of special at risk populations. Suitability involves consideration of contraindications to the medication as well as other factors such as the requirements for monitoring, drug formulation and the number of daily doses. Cost includes consideration of drug acquisition as well as the total cost of therapy, including by whom the cost is borne. PMID- 12772366 TI - Creating informed consumers and achieving shared decision making. AB - BACKGROUND: The internet has become a powerful tool to disseminate health information and there are millions of people accessing this free information for a second medical opinion. Consumers seeking information from this resource are more responsible for their own health. However, the quality of health information on the internet varies and there is no simple instrument that can effectively evaluate its quality. OBJECTIVE: This paper reviews recent studies on the validity of instruments or guidelines developed for evaluating health information. It describes government initiatives around the world in establishing quality health information and proposes an alternative way of empowering consumers with reliable health information. It emphasises the importance of the role of primary family physicians in this consumer education process. DISCUSSION: The debate over the quality of health information on the internet and the attempt to use instruments to evaluate the quality of web information are discussed. The relationship between family physicians and consumer informations in this information age is also addressed. PMID- 12772367 TI - Abdominal sepsis and communication breakdown. PMID- 12772368 TI - When things go wrong. AB - Case histories are based on actual medical negligence claims, however, certain facts have been omitted or changed by the author to ensure the anonymity of the parties involved. Despite the best intentions of medical practitioners, things do go wrong from time to time and a patient may experience an adverse outcome as a result of a medical practitioner's actions. This article discusses some of the issues that general practitioners should consider when things go wrong. PMID- 12772369 TI - Test your knowledge. Patient with a skin rash. AB - A fit and healthy 22 year old of Asian ancestry, living in New Zealand with no previous history of asthma or dermatitis, visited the Gold Coast, Queensland, for a vacation. Having been cautioned about the intensity of the Australian sun, he arrived armed with a hat and new SPF130 sunscreen that he had obtained from Japan. On the third day of his vacation he noticed the appearance of many erythematous papules and some vesicles on the dorsum of the forearm, arm and the 'v' of the neck. His face was spared. On the fourth day, the papules coalesced to form swollen, warm, very pruritic, erythematous plaques (Figure 1). The patient commenced taking loratadine 10 mg per day, but did not improve after seven days of treatment. PMID- 12772370 TI - Test your knowledge. Chronic red eye. AB - A 62 year old woman has come to the practice complaining of an irritable, red eye (Figure 1). This has occurred intermittently over a number of years. The condition tends to resolve over a week or so with the help of a series of lid bathes and hot compresses. For mild to moderate episodes she has previously been prescribed Predsol steroid eye drops for which she has responded quickly. The patient feels that the episodes are increasing and the eye is remaining 'gritty' for longer. Vision in the right eye has become poorer than the left eye. PMID- 12772371 TI - Sex in the suburbs. AB - BACKGROUND: Many suburban sexual health clinics exist in Australasia. However, sexual health research has hitherto been restricted to populations from major metropolitan centres. OBJECTIVE: To describe the demographic, sexual characteristics and presenting diagnoses of 6521 new patients attending a sexual health clinic in the western suburbs of Sydney (New South Wales). DISCUSSION: Patients originated from over 50 different countries. Postcode analysis suggested the clinic sample was representative of the socioeconomic distribution of the area, but there were notable ethnic disparities. The classic bacterial sexually transmitted infections constituted only a small proportion of presentations, with asymptomatic infections and those requiring complex therapies more commonly encountered. This has resource implications for general practitioners and specialist clinics alike. Improved liaison between specialist sexual health clinics and GPs, together with a raised awareness of sexual health services in general, has the potential to significantly improve overall public health. PMID- 12772372 TI - Cognitive behavioural therapy skills training for adolescent depression. AB - BACKGROUND: Adolescent depression is mainly managed in primary care. However, there are many problems associated with access to general practice and detection, assessment and treatment of youth depression by general practitioners. Clinical guidelines recommend the use of cognitive behavioural therapy (CBT) in treating adolescent depression. Stage 1 of the 'Time for a Future' project involved establishing a new service in Geelong, Victoria, 'The Clockwork Young People's Health Service'. The service aimed to address the barriers to optimal management of adolescent depression. OBJECTIVE: This article aims to outline Stage 2 of the Time for a Future project which involved training of 'Clockwork' GPs and other practitioners in CBT skills. DISCUSSION: The training program was well received by 68 community and Clockwork GPs, who reported significant changes in their confidence to detect, assess and treat adolescent depression. Ten GPs from Clockwork were a small but important subset of the study because the Clockwork model overcame many of the structural barriers that deter optimal management of adolescent depression by GPs. When real and perceived barriers were addressed concurrently with training, further positive outcomes were possible. PMID- 12772373 TI - 'Optimising cancer care in Australia'. AB - 'Optimising Cancer Care in Australia' is a consultative report prepared by the Clinical Oncological Society of Australia (COSA), The Cancer Council Australia (TCAA) and the National Cancer Control Initiative (NCCI). It is a blueprint for the reform of cancer care in Australia and was launched on World Cancer Day (4th February 2003). This groundbreaking document (available on the NCCI website: www.ncci.org.au) resulted from consultation with a wide range of stakeholders including consumers, health care professionals and policy makers. Themes identified by this process were developed into a workable number of key issues through a stakeholder workshop, input by a steering committee and a consultative committee, and finally by a wider process involving organisations with an interest in cancer care and reference to published evidence and international reforms. Formal input to the report was received from 140 individuals or groups, with many more having a less formal input. The report identifies key strategic steps that could make a substantial difference to cancer care, are achievable within a reasonable timeframe and at reasonable cost. These are given as recommendations, intended for early consideration, and action items intended for consideration by an implementation team or others in due course. PMID- 12772374 TI - The politics of public sector change. AB - BACKGROUND: As the changes underpinning the Coordinated Care Trials in South Australia have become more apparent, similarities have emerged between the rationalisation of public schooling in the mid 1980s and the transformation of public health in the 1990s. OBJECTIVE: This article aims to discuss the evolution of health services in South Australia and help us answer the question of how best to manage our public and private health infrastructure in a changing economic and social context. DISCUSSION: Both strategies in education and health share common elements of cost cutting, attempts at improving efficiencies, a flirting with the private sector and the attendant risk of reduced quality of services to the public. This situation in both sectors is indicative of a shift in public policy and a growth in the belief that private management of public sector infrastructure can help resolve the funding crises around our education and health systems. PMID- 12772375 TI - Priority setting and Australian general practice research. PMID- 12772376 TI - Priorities for research in the area of primary health care. How relevant are recently completed General Practice Evaluation Program projects? AB - INTRODUCTION: Seven priority areas were recently identified for research into primary health care. We report on the relevance of research funded under the previous General Practice Evaluation Program (GPEP) to the areas identified. METHOD: We analysed reports of 52 projects completed between January 1999 and September 2001. RESULTS: Projects related to the priority areas as follows: quality of care (n = 20, 38%), evidence based practice (n = 11, 21%), models of organisation and care delivery (n = 9, 17%), integration (n = 8, 15%), economic issues (n = 3, 6%), prevention and health promotion (n = 1, 2%) and health inequalities (n = 4, 8%). Thirty-two projects (57%) have implications for further research in quality of care, models of organisation and care delivery, integration, economic issues, and prevention and health promotion. CONCLUSION: Completed GPEP projects have relevance to the identified priority areas. They provide information to support research applications in the primary health care area funded through the National Health and Medical Research Council, and identify areas for further research. PMID- 12772377 TI - Training GPs to screen for diabetic retinopathy. The impact of short term intensive education. AB - OBJECTIVE: To improve general practitioners' knowledge and skills about diabetic retinopathy. SETTING: An upskilling program of 11 Sydney (New South Wales) GPs. MAIN OUTCOME MEASURES: Level of competency determined by an educational assessment before and six weeks after the program. RESULTS: Mean scores for multichoice questions increased from 53% before the education to 84% after (p < 0.001); for a photographic examination from 44 to 53% (p = 0.08); and for clinical examination from 41 to 69% (p < 0.001). The screening specificity increased from 47 to 73%. CONCLUSION: General practitioners' knowledge and skills about diabetic retinopathy improved at least short term following an intensive educational program. PMID- 12772378 TI - Should we screen diabetic patients using biguanides for megaloblastic anaemia? AB - BACKGROUND: Patients taking biguanides on a continuous basis sometimes develop vitamin B12 deficient megaloblastic anaemia. The prevalence of this side effect has not been estimated. METHODS: We screened 600 patients with type 2 diabetes treated with biguanides (phenformin or metformin) for a mean of 11.8 years (SD: 3.6 years) with complete blood counts, red cell indices and red cell morphology. If this showed macrocytosis, we measured total serum vitamin B12 and antiparietal cells antibodies (APCA). Patients with macrocytosis and low serum vitamin B12 levels were treated with cyanocobalamin 1 mg injection daily for seven days. RESULTS: Fifty-four (9%) of the patients had megaloblastic anaemia with low serum total vitamin B12 levels, only three (0.5%) also had abnormally raised APCA. All 54 patients responded to cyanocobalamin with a reticulocyte increase within 10 days. CONCLUSION: Annual screening for megaloblastic anaemia in patients on long term treatment with biguanides may be worthwhile. The anaemia is easily remediable and does not necessitate withdrawal of the drug. PMID- 12772379 TI - Bring on the substitutes. PMID- 12772381 TI - Soul survivors. PMID- 12772380 TI - Donating identity. PMID- 12772382 TI - Meet the flying eye. PMID- 12772383 TI - A little knowledge. PMID- 12772384 TI - Right to relief. PMID- 12772385 TI - Chronic fatigue syndrome: the nurse's role. AB - BACKGROUND: Chronic fatigue syndrome (CFS), a disorder of no proven cause, is characterised by extended periods of extreme, debilitating fatigue and related symptoms. This article discusses this distressing disorder and identifies the needs of those who have it. The authors suggest that the nurse could fulfil the role of key professional carer, interacting with the individual with CFS to co ordinate care and form the pivotal therapeutic relationship. CONCLUSION: CFS remains a chronic illness of uncertain cause and prognosis. For those with CFS, care priorities involve validation, information and advocacy, a therapeutic relationship and co-ordinated care. The particular philosophy of care held by nurses makes them potentially ideal co-ordinators of care for those with CFS. PMID- 12772386 TI - Improving patient hand hygiene. AB - The hand hygiene of patients has been neglected and should be emphasised. This article outlines strategies to empower patients to improve their hand hygiene, bearing in mind that they should be allowed to make their own choices. PMID- 12772387 TI - Lymphoedema. AB - Lymphoedema is a symptom of a malfunctioning lymphatic system. It is characterised by chronic swelling caused by the accumulation of lymph. Rebecca Penzer examines the nursing management of lymphoedema and the ways in which non specialist nurses can offer support and promote self-care in patients with this potentially debilitating condition. PMID- 12772388 TI - [Physicians take care of violent events themselves]. PMID- 12772389 TI - [Admission prevents violence]. PMID- 12772390 TI - [Marriages between cousins]. PMID- 12772391 TI - [Consanguinity and congenital abnormalities]. AB - Knowledge of consanguinity is relevant for employees in the Danish national health service, since about 7.5% of the Danish population has another ethnic background than Danish and the majority comes from cultures where consanguineous marriages are not unusual. In the literature it is found that consanguineous couples have a higher risk of having children with congenital malformations. The risk is increased by a factor 2 to 2 1/2. The average risk in Denmark is about 3%. Primarily, the autosomal recessive diseases are expressed in children with consanguineous parents. In order to advise and diagnose it is essential to clarify the consanguinity state. In case of pregnancy with consanguineous parents, we recommend: 1) Counselling to estimate the risk of foetal illness and information about possible examination possibilities. 2) An ultrasound scan at the gestational age of 11-14 weeks in order to measure nuchal translucency and an early malformation scan. 3) An ultrasound scan for malformations at the gestational age of 18-20 weeks. 4) An ultrasound scan especially in order to detect foetal heart malformations at the gestational age of 20-24 weeks. PMID- 12772392 TI - [Traumatic basal subarachnoid hemorrhage]. AB - Massive subarachnoid hemorrhage may occur on a traumatic basis. The injury is most often sustained by a blow with a clenched fist against the posterolateral part of the cranial basis, but the injury may also occur in relation to an accident. The condition is rare, most often occurring in alcohol intoxicated men. The victim typically collapses immediately and usually dies within a few minutes. The origin of the bleeding may be the vertebral artery on the neck or the intercranial basal brain arteries. In some cases the origin of the bleeding cannot be located. The pathogenetic mechanisms have been a subject of discussion. The damage to the artery may occur in relation to a fracture of the transverse process of the atlas or in relation to subluxations in the cervical vertebral column. The arterial rupture may occur in both normal and abnormal arteries. In many of the cases the trauma may be very slight. This has, of course, important legal implications. PMID- 12772393 TI - [Clopidogrel: background information and use in clinical practice]. AB - Acute manifestations of atherosclerosis, e.g. myocardial infarction and ischaemic stroke, are among the most common causes of death in the western part of the world. Rupture of an atherosclerotic plaque results in activation, adhesion and aggregation of platelets. Antithrombotic treatment has shown to reduce mortality and morbidity and is used in both primary and secondary prevention. The thienopyridine Clopidogrel specifically and irreversibly inhibits the binding of ADP to platelet surface ADP-receptors, thereby inhibiting platelet activation and aggregation. In this publication the pharmacological background and clinical documentation for the use of Clopidogrel both as monotherapy and in combination with Acetylicsalisylic acid are reviewed. PMID- 12772394 TI - [Dupuytren's contracture]. AB - Dupuytren's contracture is a common disease affecting the palmar fascia. It is mostly found in the elderly population, although more aggressive cases are not uncommon in the younger population. After a historic and epidemiological review, the clinical picture, histology and pathogenesis, the surgical treatment, and the latest therapeutic options are presented. PMID- 12772395 TI - [Heparin as adjuvant in the treatment of colorectal cancer?]. PMID- 12772396 TI - [Effect of Coenzyme Q10 and Ginkgo biloba on warfarin dosage in patients on long term warfarin treatment. A randomized, double-blind, placebo-controlled cross over trial]. AB - INTRODUCTION: A few case-stories claim that the anti-oxidant Coenzyme Q10 and possibly also Ginkgo biloba interact with warfarin treatment. A decreased response to warfarin in the Coenzyme Q10 cases and an increased response in the Ginkgo biloba case have been described. MATERIAL AND METHODS: Twenty-four outpatients on stable, long-term warfarin treatment were included in a randomised, double blind, placebo-controlled crossover trial. Coenzyme Q10 100 mg daily, Ginkgo-Biloba 100 mg daily and placebo were given in random order over treatment periods of four weeks, each followed by a two week wash out period. The international normalized ratio (INR) INR was kept between 2.0 and 4.0 by appropriate adjustment of the warfarin dosage. RESULTS: Fourteen women and ten men, median ages 64.5 years (33-79) were included. Three patients withdrew from the study for personal reasons. The INR was stable during all treatment periods. The geometric mean dosage of warfarin did not change during the treatment periods: Ginkgo biloba 36.7 mg/week (95% confidence interval: 29.2-46.0); CoQ10 36.5 mg/week (29.1-45.8); placebo 36.0 mg/week (28.6-45.1). CONCLUSION: The study indicated that Coenzyme Q10 and Ginkgo biloba do not influence the clinical effect of warfarin. PMID- 12772397 TI - [How many patients are on oral anticoagulant therapy in Denmark? Methods to estimate the number]. AB - INTRODUCTION: The aim of this study was to estimate the extension of oral anticoagulant therapy (OAT) in Denmark between 1997 and 1999. MATERIAL AND METHODS: 1) Data from persons who were registered at the Danish Medicines Agency with at least one prescription of a vitamin K-antagonist, and 2) persons registered in the Laboratory Information System, Aarhus County Hospital, Denmark, with three or more consecutive INR tests, with no more than four months between each test, and with at least one INR test > 1.9 were examined. RESULTS: The prevalence of OAT increased from 663 patients per 100,000 inhabitants in 1997 to 784 patients in 1999 (method 1). The prevalence increased significantly from 1997 to 1998 and from 1998 to 1999 and had a mean increase of 73.1 patients per 100,000 inhabitants (95% CI: 61.3-84.8) and a mean increase of 52.8 patients per 100,000 inhabitants (95% CI: 44.3-61.2), respectively. The prevalence of OAT was highest within the Country of Copenhagen and lowest within the County of Ringkobing. The difference between these two counties in 1997 was 0.0024 (p < 0.0001; 95% CI: 0.0017-0.0031). Within the admission area of Aarhus County Hospital (method 2), the prevalence increased from 518 patients per 100,000 inhabitants in 1997 to 630 patients in 1999. There was no statistical difference in the prevalence identified by the two methods. DISCUSSION: The study shows: 1) a significant increase in the prevalence throughout the study period, 2) a significant variation between the counties. A precise estimate of prevalence is identified by method 1. A more detailed survey is, however, provided by method 2. We recommend further investigations on the identified regional variation. PMID- 12772398 TI - [Criteria validation of the Roland Morris questionnaire. A Danish translation of the international scale for the assessment of functional level in patients with low back pain and sciatica]. AB - INTRODUCTION: The main concern of patients with low back pain is the functional limitation which the symptoms cause. Therefore it is important to find a valid tool by means of which their functional level can be assessed. The aim of this study was to validate the first Danish translation of the Roland Morris Questionnaire (RMQ), which evaluates the functional level of activity of patients with low back pain and possible sciatica. The RMQ was compared with the functional scale of a Danish functional and pain level questionnaire, The Low Back Pain Rating Scale (RS), and the functional scale (PF) of the SF-36. MATERIAL AND METHODS: Independently, three professionals made a retroversion of the RMQ. Then 135 patients with lumbar disc herniation completed the RMQ, RS and the SF 36. The patients comprised 47% women and 53% men with an average age of 43 years. RESULTS: A significant correlation between the RMQ and the other two questionnaires was found. RMQ-RS; r = 0.89, p < or = 0.001, RMQ-PF (SF-36); r = 0.88 p < or = 0.001. A Chronbach Alpha showed a value of 0.94 (scale 0-1). Differences in scores between the three questionnaires were within the 95% limits of agreement. The average RMQ score was 10% higher than the score of RS and PF (SF-36). CONCLUSION: According to this study the RMQ can be used as a valid tool in the assessment of the functional level of patients with lumbar pain and previous lumbar herniation. Due to the fact that the questionnaire is fast and easy to complete, it is valid, reliable and sensitive and widely used internationally. We recommend the questionnaire to be used as a supplement to the clinical examination both in clinical practice and in research. PMID- 12772399 TI - [Vena cava inferior atresia as the cause of deep venous thrombosis]. AB - The paper describes an 18-year-old woman with vena cava inferior atresia. She was hospitalised with deep venous thrombosis after she had taken contraceptives for two months and had been confined to bed for three weeks because of back pain. Anticoagulant treatment was successful. Vena cava inferior atresia is a rare condition, but it must be suspected in younger people with proximal bilateral deep vein thrombosis. PMID- 12772400 TI - [Cerebral venous thrombosis. Clinical presentation of three cases]. AB - Cerebral venous thrombosis is a rare but dangerous condition. We present three cases of cerebral venous thrombosis. All three patients were females. CT scan was performed. One patient had a haemorrhagic infarct in the left temporal lobe, but CT scan was normal in the last two cases. MRI later revealed that cerebral venous thrombosis was the diagnosis in all three cases. MRI is the examination of choice when cerebral venous thrombosis is suspected. The patients were treated with heparin. One developed blindness due to optic nerve atrophy. One had discreet cognitive deficits, and the last patient had no neurological sequelae after she was discharged from the hospital. PMID- 12772401 TI - [Prolonged hyperthermia after isocarboxazid poisoning]. AB - A 44-year-old unconscious man was admitted 24 hours after intake of approximately one gram of isocarboxazid. He was subsequently intubated for seven days. Muscular hyperactivity and prolonged hyperthermia were the main therapeutic problems which were treated with dantrolene, levomepromazine, pancuronium, and external cooling. Massive rhabdomyolysis was treated with forced diuresis. PMID- 12772402 TI - [Disinfection 2. A big problem in general practice?]. PMID- 12772403 TI - [A publication made in haste]. PMID- 12772404 TI - [Apropos the campaign material "It's not your fault"]. PMID- 12772405 TI - [Bilirubin induced apoptosis of human neuroblastoma cell line SH-SY5Y and affected the mitochondrial membrane potential]. AB - OBJECTIVE: To study whether bilirubin induces apoptosis of human neuroblastoma cell line SH-SY5Y and affects the mitochondrial membrane potential. METHODS: SH SY5Y cells were exposed to bilirubin, then morphological changes of nucleus were assayed by Hoechst 33258 staining. The percentage of apoptotic cells was measured by flow cytometry. The mitochondrial membrance potential(MMP) was detected by flow cytometry through Rhodamine123 staining. RESULTS: MMP of SH-SY5Y cells significantly decreased from (16.6 +/- 1.556) U to (7.11 +/- 0.701) U (P < 0.01) after exposed to 0.02 g/L bilirubin for 1 h. The percentage of apoptotic cells was 19.4% after exposed for 3 h, and significantly increased to 76.4% after 4 h, but typical apoptotic nuclear changes occured after 6 h. CONCLUSION: Bilirubin could early cause disruption of MMP, then induce the apoptosis of SH-SY5Y cells. PMID- 12772406 TI - [A report on a large family with X-linked recessive nonsyndromic hereditary low frequency neuropathic hearing impairment]. AB - OBJECTIVE: To analyze the genetic causes of low frequency neuropathic hearing impairment. METHODS: Using the network established by our institute, the proband of the low frequency neuropathic hearing loss pedigree was found. Then, investigation was carried out in the family from the proband. Cyrillic 2.1 software was set up to draw the pedigree and genetic characterization and phenotypes were analyzed in this family. RESULTS: One-hundred and one alive family members were investigated and the clinic audiologic examinations were performed in 43 of 101 individuals. Six of forty-three individuals appeared to be low frequency neuropathic hearing loss and all patients were males without systemic disorders except hearing loss. The clinic phenotypes were mild, middle, severe and profound hearing loss with disappearing of the auditory brainstem response(ABR) and partial normal results of distortion product otoacoustic emission (DPOAE) in the affected individuals. The onset of hearing loss was at 10 16 years old and the age of all patients was arranged from 18 to 26 years old. CONCLUSION: A large five generations family with hereditary low frequency neuropathic hearing impairment was found in our study. The genetic pattern in this family is male dominant X-linked recessive (XR) nonsyndromic hearing loss. Our findings suggest that some low frequency neuropathic disorders might be attributed to genetic factors. PMID- 12772407 TI - [Audiological characteristics of auditory neuropathy]. AB - OBJECTIVE: To study the auditory electrophysiological characteristics of the auditory neuropathy. METHODS: Ten patients were diagnosed as auditory neuropathy. The history, pure tone audiometry, stapedial reflex, auditory brainstem response, electrocochleogram, distortion product otoacoustic emission and his contralateral suppression of white noise, middle latency response and slow cortical response were assessed. Five patients were investigated with brain CT or MRI. RESULTS: The patients were of both genders. The average age was 20.3 years old and most of them were teenagers. Nine patients complained of bilaterally hearing impairment, while one patient who did not complain of hearing loss had a tinnitus of both ears. The pure tone audiogram showed a middle or moderate sensorineural hearing loss at low frequencies 0.5 kHz and/or 0.25 kHz in nineteen ears but a normal audiogram in one ear. Stapedial reflex was absent in nineteen ears, and threshold elevated in one ear while his pure tone audiogram revealed an ascending curve. Auditory brainstem response cannot be recorded or only wave V or/and wave I presented. All of the patients were evoked a distortion product OAE, which cannot be suppressed by contralateral white noise. Electrocochleogram demonstrated AP absent or very small amplitude in most ear and the--SP amplitude were 0.595 microV, while they had a normal slow cortical response but except one patient presented normal middle latency response. The findings of CT or MRI were normal. CONCLUSION: The pure tone audiogram could preserve unchanged at early stage of auditory neuropathy, but their stapedial reflex and auditory brainstem response are absent or threshold elevated. The precise pathologic site of this disorder is yet to be defined, but it may locate in the auditory system below brainstem. PMID- 12772408 TI - [Therapeutic trial with corticosteroid for auditory neuropathy]. AB - OBJECTIVE: To explore the feasibility of corticosteroid for the treatment of auditory neuropathy. METHODS: Six patients with auditory neuropathy, diagnosed in the first affiliated hospital of Nanjing medical university between July 1998 and December 2001, were treated with corticosteroid therapy for one month (30-40 mg of prednisone daily for the first fifteen days, and 20 mg daily for following fifteen days). Prednisone was tapered gradually if a positive response to therapy was obtained. Hearing improvement was defined as 15 dB or more threshold drops in at least two of the standard audiometric frequencies in the same ear during the trial period. RESULTS: After one month's steroid treatment, significant hearing improvement was evident in two patients with raised speech discrimination scores from 48% to 95% and from 72% to 100%, respectively. In these two cases, prednisone was used continually up to 14 and 16 weeks, respectively, and the patients still had excellent pure tone hearing abilities and speech discriminations during follow-up of three years and four months, respectively. In other four patients, positive response for corticosteroid was not found within the period of one-month trail. CONCLUSION: Immunological damages of hearing system might play part role in the pathophysiological mechanisms of auditory neuropathy, and corticosteroids are potentially useful drugs for these cases. PMID- 12772409 TI - [Changes of trkB and trkC mRNA in facial nucleus after axotomy and end-to-end anastomosis in the rat]. AB - OBJECTIVE: To explore the changes of receptor tyrosine kinase B and C (trkB and trkC) mRNA in facial motoneurons following facial nerve injury and repair. METHODS: The time course of trkB and trkC mRNA in facial motoneurons following facial nerve transection and anastomosis was analyzed using in situ hybridization (ISH) and reverse transcriptase-polymerase chain reaction(RT-PCR) in adult rats. RESULTS: In the facial nerve simple transection group, the intensity of trkB mRNA signal began to increase to 3d post-operation and to its maximum at 7d. Then, the intensity started to decrease at 14d, but it was still higher than that in the contralateral side at 35 d. After facial nerve injury, the changing trend of trkC mRNA was in the contrary to that of trkB. In the group of anastomosis, the changing trend of the intensity of trkB and trkC mRNA signal in the lesioned side was almost the same as that in the transection group. However, the intensity of trkB and trkC mRNA signal in the lesioned side returned to normal level at 35d. The finding obtained by RT-PCR was consistent with that of ISH. The statistic t test showed significant differences between the two groups for the relative density level of positive band for trkB and trkC at 21d, 35d post-operation, respectively, (ttrkB,21 = 13.795, ttrkC,21 = 7.445, ttrkB,35 = 8.560, ttrkC,35 = 10.132, P < 0.01). CONCLUSIONS: Transection of the facial nerve results in a upregulation of trkB mRNA, but a downregulation of trkC mRNA in axotomized facial motoneurons. The immediate anastomosis can't counteract and attenuate the changes of trkB and trkC induced by the nerve injury, but whether the changes of trkB and trkC return to normal level depends on the regeneration facial nerve. PMID- 12772410 TI - [Effect of staphylococcal enterotoxin B on ion electrophysiology and permeability in rabbit maxillary sinus epithelium]. AB - OBJECTIVE: To study the barrier role of staphylococcal enterotoxin B (SEB) in rabbit maxillary sinus mucosa. METHODS: Rabbit maxillary sinus mucosal membrane was dissected under a surgical microscope. The short-circuit current (Isc), conductance (G) and its permeability to horseradish peroxidase (HRP) were evaluated by employing Ussing chamber technique. RESULTS: The stimulation of SEB enhanced Isc, G and its permeability to HRP of sinus epithelial barrier. The same results were obtained by applying tumor necrosis factor alpha (TNF-alpha) to rabbit maxillary sinus. Increase in TNF-alpha level in sinus mucosa was observed by the stimulation of SEB. Those pathophysiological abnormalities of sinus mucosa were abolished by pretreatment with anti-TNF-alpha antibody. CONCLUSION: The SEB has potential effects on rabbit maxillary sinus mucosal barrier, activates the immune cells in the mucosa, and plays certain roles in maxillary sinusitis. PMID- 12772411 TI - [Immunoregulatory effect of Bacillus Calmette Guerin aerosol to allergic airway diseases]. AB - OBJECTIVE: To investigate whether bacillus calmette guerin (BCG) is effective used transairway and its preventive mechanisms to allergic airway diseases. METHODS: Animal experiment was finished. Adult rats were devided into four groups: control group, ovalbumin-sensitized group, BCG used transairway group, BCG used transairway + ovalbumin-sensitized group. Then these animals symptoms were studied and the pathology change were studied under microscope about nasal and bronchi mucosa and bronchoalveolar lavege fluid cells. IL-4 mRNA and IFN gamma mRNA in lung tissue were detected through RT-PCR, the protein production of IL-4 and IFN-gamma about bronchoalveolar lavege fluid and serum were detected through ELISA. RESULTS: In ovalbumin-sensitized group, allergic animal model were made successfully. In only BCG used transairway group, the symptoms of animals were normal, few inflammation cells infiltrated into the mucosa of nasal and bronchi, the numbers of macrophage were greatly increased in smear of bronchoalveolar lavege fluid, IFN-gamma mRNA and protein production were greatly increased. In BCG used transairway + ovalbumin-sensitized group, the allergic symptoms and inflammation were greatly reduced, not only IFN-gamma mRNA and protein production were increased but also IL-4 mRNA and protein production were greatly decreased. CONCLUSION: It is a good pathway that BCG used transairway. The immunoloregulation mechanisms of BCG to allergic airway diseases are to enhance Th1 response, in the meantime, to suppress Th2 response. PMID- 12772412 TI - [Functional and pathological study of intranasal absorption of insulin aided by azone]. AB - OBJECTIVE: To distinguish the different permeation function between sodium glycocholic acid (SGC) and Azone, and study the pathological changes induced by Azone on nasal mucosa. METHODS: Insulin was given intravenouslly at the dose of 4.0 U. The blood samples were determined by radioimmunity method. SGC or Azone was administered respectively via nasal route with Insulin 270 U/ml. Insulin nasal drops 10 microL were given to each of rabbits via nasal route. The Azone 50 microL was administered via nasal route, t.i.d., lasted for 1 week and 3 months. RESULTS: The level of blood Insulin was as follows: (3,954.47 +/- 644.47) mU/L, AUC = (1,264.87 +/- 148.00) mU.L-1.min-1 x 10(-3) in i.v. group; (740.44 +/- 94.16) mU/L, AUC = (298.02 +/- 63) mU.L-1.min-1 x 10(-3) in SGC group; (908.18 +/ 201.19) mU/L, AUC = (596.92 +/- 84.00) mU.L-1.min-1 x 10(-3) in Azone group. The bioavailability was 23.56% and 47.19% respectively. The pathological changes included cilia detachment from epithelium, glandular hypertrophy and connective tissue proliferation. CONCLUSION: It was found that Azone was an excellent permeation enhance which promoted more Insulin to be absorbed into blood stream through nasal mucosa. However, Azone also damaged the nasal mucosa in the view of pathology. PMID- 12772413 TI - [Ultraearly treatment on sudden deafness and the influence on superoxide dismutase and malonyldialdehyde of serum]. AB - OBJECTIVE: To explore the effect of treatment on sudden deafness at ultra-early stage and the influence on Malonyldialdehyde (MDA) and superoxide dismutase(SOD) of serum. METHODS: One hundred and forty-seven cases of sudden deafness (168 ears) from July 1995 to June 2001 were divided into three groups, 29 cases (36 ears) in group I treated within 6 hours after deafness, 55 cases(63 ears) in group II treated within 15 days after deafness, and 63 cases (69 ears) in group III treatment after 15 days after deafness. Total cases were treated with medicine and hyperbaric oxygen and the volume of SOD and MDA of serum were observed at the same time. There were 30 healthy volunteers as control group. RESULTS: In the group I, the hearing threshold was recovered in 18 ears, significant improved in 9 ears, improved in 7 ears and not changed in 2 ears after treatment. In the group II, the hearing threshold was recovered in 11 ears, significant improved in 16 ears, improved in 15 ears and no changed in 21 ears after treatment. In the group III, the hearing threshold was recovered in 14 ears, significant improved in 20 ears, improved in 13 ears and no changed in 22 ears after treatment. There was a significant difference in improved level of hearing between the group I and group II. (chi 2 = 9.90, P < 0.01) and between group I and group III (chi 2 = 9.30, P < 0.01). There was no significant difference between group II and group III (chi 2 = 0.03, P > 0.01). CONCLUSION: The treatment on sudden deafness in ultra-early stage is very important. The use of free radical scavenger early can protect audio nerve. PMID- 12772414 TI - [Preliminary clinical study of application of cyntridini triphosphatis and dexamethasone in treatment of sudden deafness]. AB - OBJECTIVE: To investigate the effect of application of cytridini triphosphatis (CTP) combined with dexamethasone on sudden deafness. METHODS: Two-hundred and seven patients (312 ears) with sudden deafness were randomly divided into CTP group (n = 159 ears) and control group (n = 153 ears). Besides intravenous administration of CTP, another treatments in CTP group were the same as that in control group including hyperbaric oxygen, vasodilator, energy preparation such as ATP, CoA and dexamethasone (DXM). RESULTS: The hearing threshold was (75.56 +/ 30.24) dB HL in CTP group and (72.50 +/- 40.50)dB HL in control group (P > 0.05) before treatment. The average value of the hearing threshold after treatment was decreased by (50.08 +/- 21.47) dB HL in CTP group and (19.45 +/- 19.12) dB in control group(P < 0.05), respectively. CONCLUSION: The application of CTP combined with DXM can significantly improve the effect of treatment on sudden deafness and CTP may enhance the recovery rate in the patients with sudden deafness, which was much higher than that treated by the traditional methods. However, the mechanism of the effect of CTP coupled with DXM is still unknown and further study is necessary. PMID- 12772415 TI - [Revision endoscopic sinus surgery for chronic sinusitis and polyps]. AB - OBJECTIVE: To investigate common reasons for revision endoscopic sinus surgery (RESS) and evaluate the effect of RESS. METHODS: Before the revision surgery 114 patients were examined by CT scans and nasal endoscopy, after surgery all the patients were followed up for more than 2 years. RESULTS: There were 16 cases of recurrence of polyps, 98 cases of recurrence of sinusitis. Among 114 cases there were 18 cases of anterior ethmoid sinus disease, 2 frontal recess stenosis, 31 anterior ethmoid sinuse disease with maxillary ostium obstruction, 6 middle turbinate adhesion, 18 posterior ethmoid sinus disease, 36 posterior ethmoid sinus disease with sphenoid sinus disease, 3 isolated sphenoid sinusitis. There were 15 cases with septal deviation. 99 cases were successful and 15 unsatisfied. CONCLUSION: Sinus disease that was not completely resected during the original surgical procedure due to the surgeon's less experience was the most common reason for the RESS, another reason was recurrent polyposis. The results were very rewarding except nasal polyposis. PMID- 12772416 TI - [Polygraphic characteristics of upper airway resistance syndrome]. AB - OBJECTIVE: (1) To analyze upper airway resistance syndrome (UARS) patients' polysomnographic characteristics, to lower the misdianosis of UARS. (2) To improve the understanding of UARS. METHODS: (1) Select 12 UARS patients diagnosed by PSG and continuous esophageal pressure measurement, the sleep structure, arousal index, apnea hyponea index (AHI) were analyzed, the body mass index (BMI), epworth sleepiness scale (ESS) were calculated. (2) Select 16 simple snoring cases as the control group, obstructive sleep apnea syndrome (OSAS) and UARS were ruled out by PSG and continuous esophageal pressure measurement for this group of cases. (3) Compare the 2 groups' first night PSG indexes (4) Compare the 2 nights PSG indexes of the UARS group, to see the continuous esophageal pressure measurement effects on their sleep. RESULTS: (1) There are no statistical difference of the 2 group's BMI and AHI. (2) Compared with the control group, UARS patients got higher arousal index and ESS score, more sleep stage 1 and stage 2, less sleep stage 3 and stage 4. (3) The 2 nights PSG indexes of UARS patients are not statistically different. CONCLUSIONS: (1) Compared with simple snoring cases, the UARS patients have some special characteristics, i.e., higher arousal index, less deep sleep, and higher ESS score, combined with the clinical manifestations, it is possible to screen UARS patients. (2) No evident effects of continuous esophageal pressure measurement on UARS patients sleep. PMID- 12772417 TI - [Primary culture and biologic characters of human laryngeal papillomavirus positive epithelial cells in children]. AB - OBJECTIVE: To investigate the biologic characters of HPV positive laryngeal epithelial cells in vitro. METHODS: To draw a cell growth curve by observing the biologic characters of HPV positive cells' growth pattern, PCR and nuclear acid hybridization were applied before and after culture for confirming HPV positive cells DNA. RESULTS: Primary HPV positive cells often remain viable in culture for more than six weeks, there have HPV DNA before and after culture. We found that in these cells had common characters: lag phase, growth phase and plateau phase. The cells culture showed that the first to forth day there have many cells emigration from tissue pieces. The fifth to Seventh day was latent phase, it was mainly adhere growth in this period, proliferation of culture cells wasn't active. The eighth to eighteenth day was growth phase, cells number was increased quickly, speed of growth was faster. Then cells growth entered to plateau phase, cells number was increased slowly, cells showed vacuoles obviously. It indicated that these cells came to die. CONCLUSION: Cells derives from HPV positive laryngeal epithelial have been successfully cultured, but to establish an animal model for HPV infection is difficult. PMID- 12772418 TI - [Laryngeal reinnervation for unilateral traumatic recurrent laryngeal nerve injuries]. AB - OBJECTIVE: To investigate 5 procedures of laryngeal reinnervation for unilateral vocal cord paralysis induced by traumatic recurrent laryngeal nerve injury. METHODS: 35 cases were selected for our study, all patients had unilateral recurrent laryngeal nerve injury, including 8 for nerve decompression, 6 for end to end anastomosis of recurrent laryngeal nerve, 16 for main branch of ansa cervicalis anastomosis to recurrent laryngeal nerve, 3 for nerve muscular pedicle and 2 for nerve implantation. All cases have been subjected to preoperative and postoperative voice recording, acoustic analysis, videolaryngoscopy, strobscopy and electromyography. RESULTS: It is found the adductory and abductory motion of the vocal cord restored in 5 cases with less than 4 months course who received nerve decompression. Although functional motion of vocal cord was not seen in two patients who received nerve decompression with a course longer than 4 months and one less than 4 months, and in all cases who received ansa cervicalis anastomosis and end to end anastomosis of recurrent laryngeal nerve, these procedures resulted in medialization of vocal cord and the mass and tension of the reinnervated vocal cord may become much the same as the contralateral normal vocal cord, thus resuming symmetric vibration of the vocal cords and physiological phonation. Nerve muscular pedicle technique and nerve implantation enabled adductory muscles to be reinnervated, thus improving severe hoarseness, but they didn't restore normal voice. CONCLUSIONS: (1) Nerve decompression seems to be the best procedure in laryngeal reinnervation; (2) Main branch of ansa cervicalis technique raises satisfactory reinnervation of adductor muscles; (3) Selection of the laryngeal reinnervation protocols should depend on the course, severity and type of nerve injury. PMID- 12772419 TI - [Phonomicrosurgical management of the disease of vocal fold]. AB - OBJECTIVE: To find a way of reserving normal high quality phonatory function after vocal cord surgery. METHODS: Various benign lesions (vocal nodules, polyp of vocal cord, cyst of vocal cord, etc) as well as premalignant lesion and early glottic cancer were treated with minimally excision, lateral microflap, medial microflap, submucosal infusion, mucosal stripping or epithelium stripping of phonomicrosurgical techniques to achieve minimally invasion. RESULTS: The 1,044 (99.8%) patients suffering from vocal nodules, polyp or cyst and 12 Reinke's edema achieved a normal phonation within a week. The recovery of normal phonation in 20 Reinke's edema, 31 vocal cases needed 1-3 months, by mucosal stripping. The restoration of phonation on 5 papilloma, 36 early malignancy were better than trans-cervical operation. After 5 year follow-up, 3 cases of vocal cord premalignant lesions received the re-stripping operation. 32 cases of early glottic cancer remained tumor free, 5 cases relapsed and 2 cases received the re stripping operation, and 2 cases received laryngectomy operation. CONCLUSION: The phonomicrosurgery may cure vocal disease and reserves good voice function. PMID- 12772420 TI - [Preservative surgery of T4 glottic cancer]. AB - OBJECTIVE: To explore new methods of preservative surgery for T4 glottic cancer and to evaluate their long term results. METHODS: 22 cases with T4 glottic cancer were treated surgically from 1982 to 1998. The lesions and involved cartilage and extrinsic laryngeal tissues were removed entirely, the sternohyoid myofascial flap, platysma myocutaneous flap, platysma myofascial flap, thyroid perichondral flap and hypopharyngeal mucosa flap were utilized to restore the defects of larynx, while the epiglottis or lamina of cricoid cartilage were reserved for laryngeal function restruction. RESULTS: The 3- and 5-year survival rates were 86.4% and 75.0% in all cases. 68.2% had laryngeal functions(voice, respiration and deglutition) restored and 31.8% partially restored(voice and deglutition). CONCLUSION: Despite the fact that the T4 glottic cancer usually involved with cartilage and extrinsic laryngeal tissues, preservative surgery can be carried out for the carefully selected cases with the lesions entirely removed. PMID- 12772421 TI - [Pulmonary embolism in head and neck surgery]. AB - OBJECTIVE: To improve the early diagnosis of pulmonary embolism. METHODS: A retrospective analysis was done of 5 cases of postoperative pulmonary embolism in the department of head and neck surgery from 1993 to 1999. RESULTS: 5 patients: male 4, female 1; age ranged from 42 to 64 years. The unexplained dyspnea, chest pain, and sudden death were the most frequent symptoms. 4 patients were death suddenly because of the incorrect diagnosis and treatment. Only one patient was alive for the correct diagnosis and thrombolytic treatment. CONCLUSION: Postoperative pulmonary embolism is a major cause of mortality patients undergoing head and neck surgical procedure. Mortality of pulmonary embolism could be reduced by early and correct diagnosis and treatment. PMID- 12772422 TI - [Application of rat tail collagen in patch clamp experiment with vestibular hair cells]. AB - OBJECTIVE: To study the feasibility of application of rat-tail collagen in patch clamp research in vestibular hair cells. METHODS: The effect of self-made rat tail collagen on promoting adhesion of vestibular hair cells in whole cell patch clamp experiment was observed. RESULTS: A seal was hard to be formed when the vestibular hair cells suspended among the external solution without rat-tail collagen. However, when the vestibular hair cells were firmly adhesive to the bottom of the recording chamber with rat-tail collagen, a seal can be formed easily, which fitted to the long-term observation and recording. The effect of rat-tail collagen on adhesive to vestibular hair cells is obvious. CONCLUSION: Rat-tail collagen could facilitate the vestibular hair cells to adhesive with the bottom of the recording chamber, which is helpful for long-term patch clamp research, and the collagen is considered as an optimal adhesive reagent to vestibular hair cells for patch clamp research. PMID- 12772423 TI - Dangerous ideas. Five beliefs that propel groups toward conflict. AB - The toll in death, suffering, and displacement caused by conflicts engaging groups defined by ethnicity, nationality, religion, or other social identities has reached staggering proportions over the past decade. With expertise in research and intervention, psychologists have critical contributions to make to more fully understanding and more effectively confronting this distressing global phenomenon. The authors focus on the parallels between the core beliefs of individuals and the collective worldviews of groups that may operate to trigger or constrain violent struggles. On the basis of a review of relevant literatures, 5 belief domains--superiority, injustice, vulnerability, distrust, and helplessness--are identified as particularly important for further study. PMID- 12772424 TI - Psychology, population, and reproductive behavior. AB - This special section on international perspectives in psychology focuses on population issues and reproductive behavior across national and cultural boundaries, emphasizing the responsibility of psychologists and colleagues in related disciplines to generate and communicate culturally mindful findings likely to shape public health policy and improve human well-being. After considering the historical and policy context, it highlights internationally collaborative research projects in the Czech Republic, Egypt, Mexico, and the United States that address reproductive behavior, women's roles and status, responsible parenthood, and abortion legislation in terms of cultural context, gender equity, the needs of adolescents, and public health policy. The articles illustrate the importance of the cultural context within which psychological research is conducted and how psychology can be informed by international perspectives. PMID- 12772425 TI - Reproductive behavior in the context of global population. AB - In 1900, the world's population was estimated at about 1.6 billion. With improved health and a youthful population, the world's total population more than tripled to 6 billion by 1999. There is consensus that the world's population will reach about 8.9 billion by 2050. Despite the persistent growth of population, the reproductive behavior of individuals and couples has changed significantly over the past 50 years with smaller families and lower fertility becoming the norm in many countries. Psychology not only may provide the theoretical frameworks and empirical insights into reproductive behavior but also can foster ways in which the debate on these highly personal issues can be productive for the welfare of people around the world. PMID- 12772426 TI - Being born female is dangerous for your health. AB - In this article, the author explores how gender inequity is manifested in poor reproductive and mental health outcomes, including unwanted pregnancy, unsafe abortion, maternal mortality, sexually transmitted infections, depression, and psychosomatic symptoms. Briefly described is a landmark 1994 United Nations conference emphasizing that gender inequity adversely affects women's reproductive health, particularly in developing countries, and the implementation of its recommendations is tracked. Although there is increased recognition of oppression's toll on women's physical and emotional health as well as their intellectual and social potential, progress toward equity goals is uneven and slow. Psychologists as a group play many roles--for example, in research, education, policy, law, communications, industry, international development, and private practice--through which they can make professional contributions to gender equity as a focus or underlying principle. PMID- 12772427 TI - Abortion among adolescents. AB - In most countries, adolescents' access to abortion is limited by restrictions on legal abortion. Abortion is legal in the United States, but many states require parental consent or notification. Legislation mandating parental consent has been justified by several assumptions, including high risk of psychological harm from abortion, adolescents' inability to make an adequately informed decision, and benefits of parental involvement. Empirical data raise questions about the first 2 assumptions: Studies suggest a relatively low risk associated with abortion, and adolescents seeking abortion appear to make an informed choice. Less is known about effects of parental involvement. The authors review available research and discuss policy debates over parental consent in the United States and the international context. PMID- 12772428 TI - The psychological well-being of women of Menoufiya, Egypt. Relationships with family planning. AB - Researchers surveyed the psychological well-being of 795 women of reproductive age from Menoufiya, Egypt. Five years earlier, these women had provided data relevant to their family planning behavior. This analysis links these data sets to investigate the impact of family planning on women's sense of well-being, within the context of beliefs about appropriate gender-related behaviors. Well being measures are derived for trait and state dimensions. Use of family planning and number of children born within the preceding 5 years predicted state ratings of happiness, and number of children predicted anxious pride. Neither are related to any of the trait ratings. Further, 3 different gender-role attitudes are vital to the explanation of how women define and feel good about themselves. PMID- 12772429 TI - Born unwanted. Observations from the Prague Study. AB - The authors followed 220 children born in 1961-1963 to women twice denied abortion for the same pregnancy in Prague (Czech Republic) and 220 pair-matched controls, whose mothers were pair-matched for age and socioeconomic status and the partner's presence in the home. They were medically, psychologically, and socially assessed at ages 9, 14-16, 21-23, 30, and 35. Although differences in psychosocial development between the unwanted study participants and pair-matched controls were not dramatic and changed over time, the differences were consistently in disfavor of the unwanted pregnancy participants. When siblings were used as controls, the findings supported the hypothesis that being born from an unwanted pregnancy is a risk factor for poor mental health in adulthood. PMID- 12772430 TI - Sexuality and life skills education. A multistrategy intervention in Mexico. AB - This contribution presents a set of intervention programs in the area of reproductive health, sexuality, and life-skills education that are now widely used in Mexico and with Latin American populations elsewhere. First, the authors briefly indicate how a research-informed approach has been central to the development and implementation of these interventions. Thereafter they describe how the programs were initially designed, taking into consideration the Mexican cultural context and its implications for Mexican adolescents. They also mention subsequent evaluation-based extensions of the programs to reach other target populations, to broaden the range of issues addressed, to promote their dissemination, and to advocate society-wide sexuality and life-skills education. PMID- 12772431 TI - NMCD is growing up: short thoughts on a teenager's life. PMID- 12772432 TI - Alcoholic beverage preference and characteristics of drinkers and nondrinkers in western New York (United States). AB - BACKGROUND AND AIM: Dietary and lifestyle characteristics may differ for drinkers of specific alcoholic beverages and nondrinkers which would have important implications for studies of alcohol and disease. Our aim in this study was to describe differences in dietary and lifestyle characteristics associated with alcoholic beverage preference in a population-based sample of healthy study participants. METHODS AND RESULTS: Data were collected as part of a series of case-control studies of alcohol use, myocardial infarction, and lung, breast and prostate cancer in western New York from 1846 men and 1910 women aged 35 to 79, randomly selected from the general population of Erie and Niagara Counties. Beverage preference was defined for noncurrent vs current drinkers, and drinkers of beer, wine, liquor, and mixed beverages. Generalized linear models for continuous variables and Cochran-Mantel-Haenszel statistics for categorical variables were computed for the entire sample and stratified by gender. Participant characteristics differed by alcoholic beverage preference and drinking status. In general, wine drinkers had higher education and household incomes, lower prevalence of current smoking, higher intakes of dietary fiber, potassium, vitamin E, and total carotenoids, lower total fat intakes and higher amounts of fruits, vegetables, and grain products than consumers of other beverages. Conversely, beer and liquor drinkers had somewhat lower education and household incomes, higher rates of current smoking, higher energy and total fat intakes and consumed lower amounts of fruits, vegetables, and grain products. Finally, current nondrinkers were more likely to be older, less educated, have lower household incomes, and consume diets less consistent with dietary guidelines than current drinkers. CONCLUSIONS: These results suggest that usual beverage preference may encompass other health-related behaviors and underline the importance of accurate exposure measurement and use of statistical methods to accommodate these interrelationships. PMID- 12772433 TI - Estimation of fruit and vegetable intake using a two-item dietary questionnaire: a potential tool for primary health care workers. AB - BACKGROUND AND AIM: High fruit and vegetable intake is associated with health benefits for cancer and cardiovascular disease. An increase is therefore integral in recommendations for the prevention of chronic disease. However, measuring intake requires either extensive dietary assessment or the measurement of specific bio-markers which is neither cheap nor feasible for the routine assessment of an individual's diet in a community or primary care setting. Within the context of a study evaluating a dietary counselling programme to increase fruit and vegetable intake our aim was to assess the use of a simple tool to estimate fruit and vegetable intake. METHODS AND RESULTS: We studied associations between bio-markers [plasma ascorbic acid, beta-carotene and alpha-tocopherol 24 hour urinary potassium excretion] and a two-item fruit and vegetable consumption questionnaire in 271 subjects (105 men and 166 women), aged 18 to 70 years. After controlling for age, sex, vitamin supplement use, smoking and body mass, those reporting a daily intake of > or = 5 portions of fruit and vegetables had higher potassium excretion (difference 15.6 [95% confidence interval: 6.2 to 25.0] mmol/24 h), urinary potassium/creatinine ratio (1.2 [0.5 to 2.0]) and plasma vitamin C (10.0 [-0.9 to 20.8] mumol/L) than those reporting < or = 2.5 portions per day. beta-carotene (p = 0.04), vitamin C (p = 0.01) and potassium excretion (p < 0.001) were associated with fruit rather than vegetable intake. The two-item questionnaire had high specificity; over 3/4 of participants who reported low intake also had bio-markers below the upper third of the distribution. CONCLUSION: Self report of fruit and vegetable intake through a simple questionnaire is confirmed by bio-markers for those eating less than five portions of fruit and vegetables a day. Although the tool is amenable to improvements for the detection of vegetable portions, it may prove useful for monitoring dietary preventive approaches in primary care without the use of invasive and costly biochemical measurements. PMID- 12772435 TI - Postprandial variations in the cholesteryl ester transfer protein activity, phospholipid transfer protein activity and plasma cholesterol efflux capacity in normolipidemic men. AB - BACKGROUND AND AIM: Plasma cholesterol efflux capacity is stimulated during postprandial (PP) hypertriglycerdemia. Plasma cholesteryl ester transfer protein (CETP) and phospholipid transfer protein (PLTP) are the key proteins in lipoprotein metabolism and remodelling, but their role during the PP cholesterol efflux process remains indeterminate. The aim of this study was to determine the effect of a fatty meal intake on plasma CETP and PLTP activities, and the capacity of plasma to promote cholesterol efflux, as well as to evaluate the relationship between these three key mechanisms of the reverse cholesterol transport process. METHODS AND RESULTS: CETP and PLTP activities and the cholesterol efflux capacity of plasma were measured over eight hours following a fatty meal (1000 kcal, 62% fat) in 13 normolipidemic men. CETP activity and the cholesterol efflux capacity of plasma from Fu5AH cells increased after the meal, reaching a maximum after eight hours (respectively 32%, p = 0.06, and 6.5%, p = 0.045), whereas PLTP activity remained unchanged. CETP and PLTP activities did not correlate with plasma cholesterol efflux capacity in the fasting or PP state. Plasma CETP activity in the fasting state positively correlated with the plasma non-esterified fatty acid (NEFA) levels, but no correlation was found with any lipid or apolipoprotein postprandially. The cholesterol efflux capacity of plasma correlated positively with high-density lipoprotein (HDL) components, the best correlation being with the HDL phospholipid fraction in both the fasting and PP states. CONCLUSIONS: These findings suggest that plasma CETP and PLTP activities in healthy normolipidemic subjects are differently regulated in the PP state, and are not correlated with the increased cholesterol efflux capacity of PP plasma. HDL-phospholipid remains the key factor in the regulation of the capacity of plasma to promote Fu5AH cell cholesterol efflux. PMID- 12772434 TI - Effects of long-term administration of methionine on vascular endothelium in rabbits. AB - BACKGROUND AND AIM: We studied the effects of long-term methionine administration on the vascular endothelium of Japanese white rabbits. METHODS AND RESULTS: Eleven rabbits were divided into a control group (n = 6) and a methionine-fed group (n = 5), and reared for 22 weeks. Blood samples were collected at baseline and after 22 weeks for the measurement of serum homocysteine and cysteine, serum lipids and serum superoxide dismutase activity. At the end of experiments, the animals were sacrificed, and the thoracic aorta was removed for the measurement of isometric tension and histopathological examination. The blood samples taken from the methionine group in the 22nd week showed slight but significant increases in serum homocysteine and cysteine levels (Hcy: 13.7 +/- 1.4 vs 21.0 +/ 4.9, p < 0.01; Cys: 241.6 +/- 37.8 vs 342.6 +/- 35.0, p < 0.01). In the isometric tension experiments, the methionine group had a significantly decreased (p < 0.01) vasodilatation reaction induced by acetylcholine, an endothelium dependent vasodilator. The histopathological examination (immunostaining in response to eNOS and tissue factor) showed significant increases in endothelium expression in the methionine group before atherosclerotic changes appeared. CONCLUSIONS: The above results suggest that vascular endothelial dysfunction played an important role in the atherosclerosis occurring after excess methionine feeding. PMID- 12772436 TI - Histomorphometric, biochemical and ultrastructural changes in the aorta of salt loaded stroke-prone spontaneously hypertensive rats fed a Japanese-style diet. AB - BACKGROUND AND AIM: It is demonstrated that dietary habits play a role in cardiovascular diseases. In stroke-prone spontaneously hypertensive rats (SHRsp), concomitant salt loading and a Japanese-style diet greatly accelerate hypertension and the appearance of cerebrovascular lesions by directly damaging arterial vessels. A number of studies have characterised medium and small vessel lesions in SHRsp, but little attention has been paid to the changes in the wall structure of large arteries induced by exposure to a salt-enriched diet. The aim of this study was to investigate the effects of a Japanese-style diet and salt loading on the thoracic aorta. METHODS AND RESULTS: Two-month-old SHRsp were kept on a Japanese-style diet with 1% sodium chloride solution replacing tap water. Two months later, they were sacrificed and compared with age-matched or two-month old control SHRsp kept on a standard diet and tap water in terms of the histomorphometry, ultrastructure and biochemical composition of the thoracic aorta. The vessel was consistently thicker in the four-month-old SHRsp (+20%, p < 0.05 vs two-month-old rats) regardless of diet. The salt-loaded SHRsp showed a significant reduction in elastic fibre density (-20%, p < 0.05 vs two-month-old rats) and an increase in the other matrix components (%), whereas the four-month old controls showed preserved elastic fibres and a significant increase in the other matrix components (+65%, p < 0.05 vs two-month-old rats). There was a considerable increase in the amounts of 4-OH-proline (+147%), 5-OH-lysine (+174%) and desmosines (+360%) in the four-month-old controls vs their two-month-old counterparts (p < 0.01), but not in the salt-loaded animals. Ultrastructural analysis revealed clear damage and accelerated aging in the thoracic aorta of the salt-loaded SHRsp. CONCLUSIONS: Salt loading and a Japanese-style diet destabilize thoracic aorta architecture in SHRsp after two months of treatment. PMID- 12772437 TI - Plasma lipoproteins, apoproteins and cardiovascular disease in type 2 diabetic patients. A nine-year follow-up study. AB - BACKGROUND AND AIM: To evaluate the role of lipoprotein abnormalities as risk factors for macroangiopathy in Type 2 diabetes. METHODS AND RESULTS: This prospective nine-year follow-up study involved 113 Type 2 diabetic patients (50 men and 63 women, mean age 66.9 +/- 9.9 years), 37 of whom had clinical signs of coronary heart disease (CHD) and cerebrovascular disease (CVD) at baseline. During the follow-up, 32 patients died: 17 of CHD, five of CVD, and 10 of non vascular causes. The patients who died because of vascular disease were more frequently smokers, and had baseline symptoms of vascular disease; they were also significantly different from the other patients insofar as they were older, and had higher fasting plasma glucose levels, lower fasting C-peptide levels, and lower apoprotein (apo) AII, apo CII, apo CIII and apo E levels. Univariate analysis showed that baseline symptoms of vascular disease, current smoking, age, high fasting plasma glucose levels, low fasting C-peptide levels, and low apo AII, apo CII, apo CIII and apo E levels [but not cholesterol, triglyceride, high density lipoprotein (HDL)-cholesterol or qualitative low-density lipoprotein or HDL abnormalities] were associated with cardiovascular mortality. Multivariate analysis showed that only age, smoking, glycated hemoglobin (HbA1c) and fasting C peptide levels were significant independent determinants of macrovascular death. CONCLUSIONS: In Type 2 normolipidemic diabetic patients, only age, smoking, HbA1c and fasting C-peptide levels are independent vascular risk factors. The differences in apo concentrations between patients with and without vascular disease may reflect qualitative abnormalities in plasma lipoproteins related to vascular disease. PMID- 12772438 TI - The psychosocial determinants of hypertension. AB - AIM: To present currently available evidence on the role that adverse psychosocial factors play in the pathogenesis of hypertension. The specific objectives of the review were to (1) provide a picture of what is known about the relationship between psychosocial factors and hypertension, (2) summarize the major methodological and conceptual pitfalls, and (3) identify gaps in the literature and suggest areas for future research. DATA SYNTHESIS: The scope of the literature review was adults and the literature published since 1990 (acknowledging that some articles published earlier would need to be taken into account). A number of journal searches were carried out. They included Medline, PsychInfo, and SocioFile, with keywords, such as hypertension, blood pressure, psychosocial, psychological, social, acculturation, occupation, socio-economic status, social class, education, depression, anger, and anxiety. The search included articles related to hypertension in developed countries and was limited to studies in the English language. CONCLUSION: A growing body of evidence supports the thesis that psychosocial factors play a role in the pathogenesis for hypertension. PMID- 12772439 TI - WHO Expert Committee on Drug Dependence. AB - This report presents the recommendations of a WHO Expert Committee responsible for reviewing information on dependence-producing drugs to assess the need for their international control. The first part of the report contains a description of the procedure for the review of dependence-producing substances. This is followed by a critical review of five psychoactive substances (amfepramone, amineptine, buprenorphine, delta-9-tetrahydrocannabinol and tramadol). The report also discusses the substances that were pre-reviewed by the Committee, four of which (ketamine, zopiclone, butorphanol and khat) were recommended for critical review at a future meeting. The final section discusses the problems of the terminology used in reporting abuse-related adverse drug reactions and describes how confusion affects the reporting of adverse effects using the example of the selective serotonin reuptake inhibitors. PMID- 12772440 TI - Small employers and health benefits: findings from the 2002 small employer health benefits survey. AB - Overall, 19 percent of small employers offering health benefits made changes to their health plan between 2001 and 2002. Sixty-five percent increased deductibles and co-pays; 35 percent switched insurers; 30 percent increased the employee share of the premium; and 29 percent cut back on the scope of benefits. Twenty six percent increased the scope of benefits offered. Nearly one-quarter of small employers offering health benefits think their firm would change coverage and 3 percent think it would drop coverage if the cost were to increase an additional 5 percent. Most small employers offer sound business reasons for offering health benefits to workers. Many report that it helps with employee recruitment and retention, and increases productivity. More than three-quarters report that offering health benefits is "the right thing to do." Most small employers that do offer health benefits report that it has a positive impact on various aspects of the business, such as recruitment, retention, employee attitude and performance, employee health status, and the overall success of the business. Most small employers that do not offer health benefits tend to think that not offering them has no negative impact on the above aspects of their business or the overall success of the business. However, those not offering benefits are more likely than those offering them to report that most of their employees are high-turnover and stay on the job only a few months. Small employers that offer health benefits tend to be distinctly different from those not offering them. Worker income in firms not offering health benefits tends to be considerably lower than in firms that do offer them. Employers not offering health benefits are more likely than those offering them to have a smaller proportion of full-time employees, and employers that do not offer health benefits have a larger proportion of females, workers under age 30, and minority employees. Of small employers that offer dependent coverage, more than 40 percent report that workers do not take coverage for their dependents because the dependents have coverage from somewhere else, but 35 percent report that employees decline dependent coverage because they cannot afford the premiums. Many small employers that do not offer health benefits are potential purchasers. Eleven percent are either extremely or very likely to start offering health benefits in the next two years, and 22 percent are somewhat likely to start offering health benefits. PMID- 12772442 TI - Gender-role attitudes among Egyptian adolescents. AB - Using nationally representative survey data, this study explores gender-role attitudes among unmarried adolescents aged 16-19 in Egypt, a society characterized by distinct and often segregated roles for men and women. Adolescents' views about desirable qualities in a spouse as well as more direct indicators of gender-role attitudes are examined, including opinions about whether wives should defer to their husbands, share in household decision making, and have the responsibility for performing domestic tasks. The findings regarding spousal characteristics reflect strong gender differentiation. Girls and boys provide divergent profiles of an ideal spouse, profiles that reflect traditional gender roles. Girls are significantly less likely than boys to favor educational inequality between spouses, however. Neither boys nor girls have egalitarian gender-role attitudes, although girls are significantly more likely to express less traditional attitudes. Multivariate analyses indicate that girls' and boys' attitudes do not vary consistently and significantly by socioeconomic background; in particular, increased schooling does not always promote egalitarian attitudes. The implications of these findings are discussed. PMID- 12772441 TI - The proximate determinants of the decline to below-replacement fertility in Addis Ababa, Ethiopia. AB - Between 1990 and 2000, the total fertility rate (TFR) in Ethiopia declined moderately from 6.4 to 5.9 children per woman of reproductive age. During the same period, the TFR in the capital city of Addis Ababa declined from 3.1 to 1.9 children per woman. Even more striking than the magnitude of this decline is that it occurred in the absence of a strong and effective national family planning program. In this study, the components of this fertility decline are identified using the Bongaarts framework of the proximate determinants of fertility. The results of a decomposition analysis indicate that a decrease in the age-specific proportions of women who are married, followed by an increase in contraceptive use are the most important mechanisms by which fertility has declined in Addis Ababa. Poor employment prospects and relatively high housing costs are likely factors that encourage couples to delay marriage and reduce marital fertility. PMID- 12772443 TI - Associations of mass media exposure with family planning attitudes and practices in Uganda. AB - This study examines the associations between multimedia behavior change communication (BCC) campaigns and women's and men's use of and intention to use modern contraceptive methods in target areas of Uganda. Data are drawn primarily from the 1997 and 1999 Delivery of Improved Services for Health (DISH) evaluation surveys, which collected information from representative samples of women and men of reproductive age in the districts served by the DISH project. Additional time trend analyses rely on data from the 1995 Uganda Demographic and Health Survey. Logistic regressions are used to assess the associations between BCC exposure and family planning attitudes and practices, controlling for individuals' background characteristics. To minimize the biases of self-reported exposure, the analyses also explore cluster-level indexes of the penetration of BCC messages in the community. Results indicate that exposure to BCC messages was associated with increased contraceptive use and intention to use. Some evidence of self-reported bias is found, and the pathways to fertility-related behavioral change appear different for women and men. PMID- 12772444 TI - Inequality and changes in women's use of maternal health-care services in Tajikistan. AB - Using recently available survey data for Tajikistan, this study explores changes in the pattern of maternal health care during the last decade and the extent to which inequalities in access to that care have emerged. In particular, the links between poverty and women's educational status and the use of maternal health care services are investigated. The survey findings demonstrate a significant decline in the use of maternal health-care services in Tajikistan since the country gained independence from the Soviet Union in 1991. They show changes in the location of delivery and the person providing assistance, with a clear shift away from giving birth in a medical facility toward giving birth at home. More than two-fifths of all women who gave birth in the year prior to the survey delivered their baby at home. Women from the poorest quintile are three times more likely than women from the richest quintile to undergo a home delivery without a trained assistant. PMID- 12772445 TI - Contraceptive use before and after marriage in Shanghai. AB - Data from a cohort study of 7,336 newly married fertile couples conducted between 1987 and 1995 were used to analyze contraceptive method choice, switching, and discontinuation in two districts of Shanghai. Twelve percent of couples reported that they had had sexual intercourse before marriage. Only one-third of those exposed to premarital risk of conception were protected by some form of contraception, mostly by withdrawal and periodic abstinence. As a consequence, a majority of these couples conceived, prompting rapid marriage in most cases and induced abortion among one-fourth of them. After marriage, about half of the couples used contraceptives to postpone the birth of their first child, but of these, 40 percent experienced an unintended pregnancy. Method choice was dominated by condoms, withdrawal, and abstinence. After the birth of their first child, almost all couples (98 percent) adopted contraceptives, but one-third of them used ineffective methods. Failure and discontinuation rates were high, giving rise to a high incidence of induced abortion. Increasing numbers of couples switched to the IUD, and this was the preferred method for the majority by the third year following childbirth. These results suggest that wider method choice that includes hormonal contraceptives should be provided to meet couples' needs in Shanghai and that the family planning program's attention should be focused specifically on sexually active unmarried individuals and on the availability of postpartum services. PMID- 12772446 TI - Cote d'Ivoire 1998-99: results from the Demographic and Health Survey. PMID- 12772447 TI - Turkmenistan 2000: results from the Demographic and Health Survey. PMID- 12772448 TI - [Distribution of L-type calcium channels on outer hair cells among various turns of guinea pig cochlea]. AB - OBJECTIVE: To probe the relationship between the density of L-type calcium channels on outer hair cells(OHCs) among various turns and the tonotopic map in the guinea pig cochlea. METHODS: The L-type calcium channels in outer hair cells of guinea pigs were labeled with DM-BODIPY DHP and RH414 and monitored using a laser scanning confocal microscope. The fluorescence ratio of DM-BODIPY DHP/RH414 revealed the distribution of L-type calcium channels on the cellular membrane. RESULTS: The results showed that the cellular membrane had lesser DM-BODIPY DHP staining when pretreated with nifedipine, indicating that calcium channels on OHCs had the specificity of a L-type calcium channel. The density of L-type calcium channels had a tendency of increasing gradually from low frequency areas to high frequency areas. Variance analysis showed that the difference of the density of L-type calcium channels on OHCs between the fourth turn and the second turn was significant. There were no apparent differences in the distribution of L type calcium channels among various regions on the cellular membrane of OHCs. CONCLUSIONS: The descripency of the distribution of L-type calcium channels on OHCs among various turns of the guinea pig cochlea may be one of the molecular mechanisms of the tonotopic map of the cochlea. PMID- 12772449 TI - [Effect and its mechanism of gentamicin on melanin in stria vascularis of guinea pig]. AB - OBJECTIVE: To investigate the effect and its mechanism of gentamicin(GM) on melanin in stria vascularis of guinea pig. METHODS: The differences of auditory thresholds between pigmented and albino guinea pigs, given GM of 150 mg/kg for 7 days, were studied. Moreover, the content of melanosomes, activity of tyrosinase and expression of proliferating cell nuclear antigen(PCNA) in intermediate cells of stria vascularis in gentamicin-treated pigmented guinea pigs were compared with those in control animals by electron microscope and immunohistochemistry, respectively. RESULTS: After gentamicin exposure, the auditory thresholds of all animals increased significantly (P < 0.001), whereas threshold shifts averaged across all frequencies of pigmented animals were much less than those of the albinos(P < 0.001). The number of melanosomes of each examined area (300 microns 2) in intermediate cells was obviously increased from 19.83 +/- 2.74 to 58.33 +/- 16.22. The ratio of tyrosinase reaction products area to the total measured area was significantly increased from 1.65% +/- 0.40% to 3.45% +/- 0.41% after gentamicin exposure. However, the numbers of positive intermediate cells expressing PCNA were 14.08 +/- 2.76 and 13.58 +/- 2.09 before and after gentamicin treatment, respectively. But there was no statistically significant difference between them (P > 0.05). CONCLUSION: The increase of content of melanin in stria vascularis after GM exposure does not result from the change of proliferating activity of melanocytes, but from the enhanced tyrosinase activity. Melanins in stria vascularis may possess the ability to protect the inner ear from ototoxicity of gentamicin. PMID- 12772450 TI - [Tissue-engineered auricled cartilage: an experimental study]. AB - OBJECTIVE: To study the feasibility of engineering auricled neocartilage with chitosan/polylacticacid-polycrylactone (PLA-PCL) network scaffolds and to search the difference between dynamic and silent chondrocytal culture techniques. METHODS: Chondrocytes from auricled cartilage of 4 weeks old New Zealand White rabbit were seeded onto chitosan/PLA-PCL network. Ten cell-polymer scaffolds were divided into two groups: dynamic group (n1 = 5) with rotating bioreactors and silent group (n2 = 5) with ordinary dishes to culture. Using scan electroscope, grossly histological and immunohistological techniques, the morphological evaluation was done individually at 1st week in vitro, 4th and 8th week in vivo. RESULTS: Chondrocytes adhered and grew up well on the network, but more quantities of chondrocytes, Glycoaminoglygan (GAG) and type II collagen were found in dynamic group. There was an obvious difference between dynamic and silent group (P < 0.05). CONCLUSION: Chitosan/PLA-PCL network scaffold is good for adhesions and growth of chondrocytes. Furthermore, dynamic cell culture method is better than silent method for formation of neocartilage. PMID- 12772451 TI - [The neuroattract growth effect of ciliary neurotrophic factor on recurrent laryngeal nerve regeneration]. AB - OBJECTIVE: To investigate the neuroattract growth effect of Ciliary neurotrophic factor (CNTF) on recurrent laryngeal nerve (RLN) regeneration. METHODS: 28 canines were divided into 4 groups (n = 7). RLN was severed and sutured in silicone chamber. CNTF or saline were injected into silicone chambers during nerve repair and injected subcutaneously every day after surgery in group A and B respectively. In group C and D, end to end anastomosis of RLN was performed, and CNTF or saline was used respectively for one month. The nerve regeneration and neuroattract growth effect were evaluated with electrophysiological, morphological examination and horseradish peroxidase(HRP) retrograde tracing 2, 3 and 4 months postoperatively. RESULTS: Physiological adductory and adductory motion was observed in all 7 cases in group A and C respectively, and in 3 cases in group B. No motion in group D. Misdirected electrical activity by misdirected nerve regeneration was found in 3 cases in group B and 6 cases in group D. On average, the potential of misdirected electrical activity in group D was significantly greater than that in group B (P < 0.05). Compared with group B, the number of HRP retro-gradely labeled motoneuron increased significantly in group A in ambiguous nucleus, but in nodosity shape ganglia the number of the labeled neurons were not significantly different between group A and B. The average number and diameter of myelinated axons in the group A and C were significantly greater than those in group B and D, but no significant difference was found between group A and B, D and C respectively. CONCLUSION: The results suggested that exogenous CNTF can enhance motor axon regeneration and have neuroattract growth effect on recurrent laryngeal nerve regeneration. PMID- 12772452 TI - [Mitochondrial DNA mutations and non-syndromic sensorineural hearing loss]. AB - OBJECTIVE: To analyze the incidence of three types of mitochondrial DNA mutations in the non-syndromic sensorineural hearing loss (NSSNHL) patients and control subjects in order to investigate the possible role of mitochondrial DNA mutations in NSSNHL. METHODS: Sixty-one sporadic NSSNHL patients (from 3 to 84 years old) and 19 control subjects matched for age were selected. DNA was extracted from isolated blood leukocytes. Interrupt polymerase chain reaction (PCR) and primer shift PCR were used to detect the mtDNA4977 deletion; mtDNA1555A-->G and mtDNA3243A-->G point mutation were detected by PCR and restriction fragment length polymorphism (RFLP) analysis. PCR products were sequenced by automated laser fluorescent DNA sequencer. RESULTS: The detection rate of mtDNA4977 deletion in deafness groups and control groups are 68.85%(42/61) vs. 5.26%(1/19). Among all the samples, neither any mtDNA1555A-->G mutation nor mtDNA3243A-->G point mutation was detected. CONCLUSIONS: MtDNA4977 deletion had a high detection rate in patients with NSSNHL. MtDNA1555A-->G mutation and mtDNA3243A-->G point mutation may not be common mutations in patients with NSSNHL. PMID- 12772453 TI - [KCNQ4 gene mutations affected a pedigree with autosomal dominant hereditary hearing loss]. AB - OBJECTIVE: To investigate if the KCNQ4 gene contributes to a Chinese non syndromic hearing loss pedigree and to detect the gene mutations in the pedigree using candidate approach. METHODS: PCR-SSCP and clone sequencing were performed to identify the mutations and polymorphism in PCR products of KCNQ4 coding sequence in the six-generations pedigree of autosomal dominant hereditary hearing loss. RESULTS: Mutations and polymorphism detection were performed on the KCNQ4 coding sequence in 36 family members of the pedigree. A molecular polymorphism marker located in the exon2 and exon3 intron sequence, which resulted from a copy variation of 47 base pairs insertion or deletion, was found in KCNQ4 sequence. CONCLUSION: A new molecular polymorphism marker with different genotypes was proved to locate at the intron sequence between at exon2 and exon3. The correlation between genotype and phenotype was analyzed. Deaf individuals were accompanied by the increase of the intron copies in the family. These findings suggest that the changes of the copies of intron between exon2 and exon3 of KCNQ4 might be a specific marker for the hearing loss of the pedigree. PMID- 12772454 TI - [Mutation analysis of Cx26 gene in Chinese hereditary nonsyndromic deafness sufferers]. AB - OBJECTIVE: To analyze the mutations in the code region of Cx26 gene in Chinese hereditary nonsyndromic hearing impairment (NSHI) sufferers. METHODS: Thirty three cases (29 cases in the families of 8 students who were picked out from the Deafness and Muteness School of Tianjin, 2 cases as control and 2 normal cases of genetic counseling) were included in this study. The blood samples were obtained to distill the DNA templates. Using polymerase chain reaction (PCR), the code region of Cx26 gene was amplified. The mutations were screened by restriction endonucleases fingerprinting-single strand conformation polymorphism (REF-SSCP). Afterwards we inspected the polymorphous changes or mutations of these segments with DNA sequence. RESULTS: There were 30 cases with the nucleotide changes in the Cx26 code region. The rate was 90.9% (30/33). Eight kinds of mutations were found, 79G-->A, 109G-->A, 161A-->T, 235delC, 240G-->A, 341A-->G, 571T-->C and 608T-->C. 161A-->T, 240G-->A and 571T-->C were detected primarily. There were 3 cases with 235delC in 22 deafness sufferers and the rate was 13.64% (3/22). CONCLUSIONS: The 235delC of Cx26 gene is the main mutation in Chinese NSHI sufferers, and in NSHI cases many polymorphous changes exist. PMID- 12772455 TI - [The diagnosis and treatment of primary tumors of facial nerve: a report of 14 cases]. AB - OBJECTIVE: To summarize the clinic characteristics of the tumors of facial nerve and discuss the diagnosis and treatment. METHODS: Fourteen cases of primary facial nerve tumors confirmed pathologically and treated in the Chinese PLA General Hospital from 1986-2000 were reviewed. The clinic manifestation, diagnosis and treatment in this series were analyzed. RESULTS: Among the 14 cases, there were 9 of facial neurilemmoma, 3 of facial neurofibroma and 2 of facial nerve hemangioma. The commonest first symptom was facial paralysis (10/14), the next was hearing loss(6/10). The commonest sign was the facial paralysis. The following was tumor in tympanic cavity (5/14) and the next was tumor in external auditory canal(4/14). The tumors in all 14 cases were totally resected surgically. The function of the facial nerve was normal in one case with nerve being preserved intact during operation, and in grade II in one case and grade III in other one case with nerves being preserved partly continuous. Facial -great auricular-facial nerve cable grafting were done in 6 cases, as result their facial nerve were in grade II in 2 cases, grade III in 3 case and grade IV in 1 case. The facial-lateral femoral cutaneous-facial nerve cable grafting was done in one case, and the facial nerve function was in grade III with 6 years of following-up. The second-stage facial-hypoglossal nerve anastomosis was done in one case, and the facial function consequently recover to grade II with 3 years of following-up. Three cases of facial nerve remained discontinuity and their facial nerve function showed no recovery (in grade VI). CONCLUSIONS: Once the facial nerve tumors were diagnosed, the surgical treatment should be done as early as possible. At the same time of tumor resection, the continuity of facial nerve should be kept and reconstructed in one stage as possible as we can, otherwise the second-stage surgery have be conducted soon after. PMID- 12772456 TI - [Expression of P-selectin in nasal polyps and its mechanisms]. AB - OBJECTIVE: To investigate the expression of P-selectin and its mechanism in nasal polyps. METHODS: The expression of P-selectin was detected with immunohistochemistry and reverse transcriptase polymerase chain reaction (RT-PCR) in nasal polyps from 24 patients and nasal epithelium from 9 normals. The production of P-selectin in supernatans of isolated cultured vascular endothelial cells induced by interleukin-5 (IL-5) was detected with enzyme-linked immunosorbent assay (ELISA). RESULTS: The positive rate of P-selectin in nasal polyps was 79.1% (19/24) while it was 22.2%(2/9) in normals. The relative density ratio of P-selectin and beta-actin were 0.68 +/- 0.11 in nasal polyps and 0.29 +/ 0.08 in normals indicated by RT-PCR and agrose electrophoresis. The concentration of P-selectin was (53.4 +/- 2.9) ng/L and (21.3 +/- 3.5) ng/L in the two groups respectively. CONCLUSIONS: P-selectin upregulated in nasal polyps correlated with the infiltrated eosinophils and IL-5 could induce the expression of P-selectin protein in the isolated cultured endothelial cells. It suggested that upregulation of P-selectin may play a role in the pathogenesis of nasal polyps and can be enhanced by cytokine IL-5. PMID- 12772457 TI - [Orbital surgery by transnasal endoscopic ethmoidal-lamina papyracea approach]. AB - OBJECTIVE: To explore the feasibility of transnasal endoscopic surgery to treat some kinds of diseases within orbit by ethmoidal-lamina papyracea approach, and put forward the basic principle of surgical management. METHODS: A total of 10 cases undergone the endoorbital operation by transnasal endoscopic ethmoidal lamina papyracea approach, including foreign body 4 cases, sponge haemangioma 1 case, invaded and metastatic carcinoma 5 cases. RESULTS: Successful removing of foreign body from endoorbital for 3 cases, and 1 failure. One case of sponge haemangioma was removed completely. One case of metastatic tumour from nasopharyngeal carcinoma was resected, with no relapse and vision recovered to 0.6 during follow-up period of 4 years. Four cases of invaded carcinoma from ethmosphenoidal sinus were resected completely. Follow-up for 1-4 years showed no relapse. CONCLUSION: The endoorbital region inside the optic nerve can be controlled by transnasal endoscopic ethmoidal-lamina papyracea approach. The advanced radiotherapy provides favourable conditions for conservative surgery of endoorbital carcinoma. PMID- 12772458 TI - [Endoscopic intrasellar tumor surgery]. AB - OBJECTIVE: To report the experience with the perioperative management of endoscopic intrasellar tumor surgery in fifty-two patients with intrasellar tumor who underwent an endoscopic transsphenoidal approach to the sella. METHODS: The application of transnasal endoscopic technique to the management of pituitary adenomas in 49 patients, of craniopharyngioma in two patients and of meningeoma in one patient. RESULTS: Forty-eight of forty-nine cases with pituitary adenomas and two cases with craniopharyngioma were removed completely, the majority of one case with invasive pituitary adenoma and the meningeoma were removed on purpose to decompress. The symptoms of headache, visual desturbance, amenorrhea, galactorrhea ect. were cured or improved. The endocrinological evaluation: the high preoperative prolactin (PRL) and growth hormone (GH) level were decreased to normal values after the operation. Transient and permanent diabetes insipidus occurred in five patients and one patient with pituitary adenomas respectively after the operation, only one case required the treatment. One patient with craniopharyngioma had postoperative epilypsy. Two patients with pituitary adenomas and one patient with craniopharyngioma had recurrence within the one year (recurrence rate 5.8%) during the follow-up 3-72 months after the surgery. No patient developed a delayed cerebrospinal fluid leak, meningitis, optic nerves injury, internal carotid injury, or other endocrine complications. CONCLUSION: This new technique offers simple and rapid access to the sella turcica and also, direct visualization, improves the surgeons ability to resect tumors, it is a safe, minimally invasive and efficient procedure for removing intrasellar tumor. PMID- 12772459 TI - [Surgical management of cerebrospinal fluid rhinorrhea]. AB - OBJECTIVE: To summarize the clinical experience in managing cerebrospinal fluid (CSF) rhinorrhea. To ascertain the best effective surgical management for CSF rhinorrhea. METHODS: A retrospective study of 83 patients with CSF rhinorrhea from March 1991 to August 2001 was conducted. Among them, 17 patients were treated with conservative therapy, 8 with transcranial repair, 2 with extranasal repair, 3 with transnasal repair under microscope and 54 with transnasal endoscopic management. The conservative time, causes, complications, leak sites, repairing materials and surgical methods were analysed. RESULTS: All patients were followed -up for 6 months to 9 years. 17 cases had no CSF rhinorrhea after conservative treatment with an average time 16 days. Endoscopic repair of CSF leaks was successful in 90% (49/54) of the cases after a first attempt. Two persistent leaks (2/4) were closed after a second attempt. The other persistent leaks were also closed, one case after third and one after fourth attempt. Ultimately 98% (53/54) of the leaks were repaired using an endoscopic approach. Only one failed after transendoscopic management, and succeeded with a second extranasal repair. The CSF rhinorrhea repaired with other surgical methods were all successful. CONCLUSION: Transnasal endoscopic management is the best method for repairing CSF rhinorrhea, other surgical methods can be chosen based on different conditions. The conservative time are 2 to 4 weeks if a CSF leak is caused by trauma or operation, 6 to 8 weeks conservative time for special cases. PMID- 12772461 TI - [Genetic polymorphism of cytochrome P4501A1 and susceptibility to laryngeal carcinoma]. AB - OBJECTIVE: To assess the possible relation between the MspI polymorphism of Cytochrome P4501A1(CYP1A1) gene and the susceptibility to laryngeal cancer. METHODS: The genotypes of CYP1A1 MspI site were detected using the methods of polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) in 62 cases of laryngeal squamous carcinoma and 56 healthy controls. Genetic risk of CYP1A1 genotypes was analyzed by smoking index (SI, cigarettes smoked per day x years of smoking). RESULTS: Three genotypes of CYP1A1 MspI were classified into the predominant homozygotes (A), heterozygotes (B) and the rare homozygotes (C). The frequency of heterozygote B (58.1%) and genotype C (14.5%) in the patients with laryngeal cancer were higher than that of the controls (39.3%, 7.1%, P < 0.05), while their odds ratios were 2.89 (95% confidence interval, CI: 1.31-6.37) and 3.97 (95% CI: 1.10-14.28), respectively. The odds ratios of genotype C was 9 (95% CI: 1.60-50.74) in the high dose cigarette smoking group, but was 4.5 (95% CI: 0.64-31.61) in the low dose cigarette smoking group. CONCLUSIONS: With the carcinogenesis and development of laryngeal cancer, the polymorphism of CYP1A1 gene and smoking exposure together may play an important role. The individuals with genotype C are at especially high risk of laryngeal cancer, which grows with increasing cigarette consumption. PMID- 12772460 TI - [Reconstruction of anterior skull base defect by pedicle frontal muscle compound flap]. AB - OBJECTIVE: To evaluate the effect of one stage reconstruction for anterior skull base defect by pedicled frontal muscle compound flap after craniofacial combined approach operation. METHODS: Twelve patients with frontal skull base tumor and fracture (rudimentary cancer 5, malignant melanoma 1, neurogliocytoma 1, ethmoid sinus rhabdomyosarcoma 1, malignant papilloma 1, osteofibroma 2, underwent surgery 1) were included. The compound flap with pedicled frontal muscle galea aponeuroses pericranium was adopted which brought single blood vessel or double, and splinter skull bone. The compound flaps covered 8 cm -12 cm x 10 cm -15 cm. RESULTS: All 12 patients were successfully treated with no complication during follow-up from 1 to 48 months. CONCLUSION: The compound flaps with pedicled frontal muscle galea aponeuroses pericranium, had ample blood supply and thin pliable and strong tissue which was a good reconstruction material for frontal skull base defect. PMID- 12772462 TI - [Rib cartilage graft laryngotracheal reconstruction for severe laryngotracheal stenosis]. AB - OBJECTIVE: To study the success and failure factors with rib cartilage graft laryngotracheal reconstruction for severe laryngotracheal stenosis for improving the rate of successful decannulation. METHODS: Thirty-six cases with severe laryngotracheal stenosis, received rib cartilage graft for laryngotracheal reconstruction during the last 18 years in our hospital, were analyzed retrospectively. RESULTS: Twenty-nine of the 36 patients (80.5%) were successfully decannulated after one procedure. The follow-up time ranged from 1 to 10 years. All of the 29 patients had a stable airway and normal exercise tolerance. The duration of T-tube stent was 10 days to one year (average 3 months). Seven patients (19.5%) failed to decannulate. The factors of failure were disturbance of local blood circulation, wound infection followed rib necrosis, T-tube injured mucomembrane then became a new stenosis, and restenosis for scar constitution. CONCLUSION: Laryngotracheal reconstraction with rib cartilage graft is one of effective method for severe laryngotracheal stenosis. The procedure is relatively simple, and shorter duration of stenting and higher rate of success. The drawbacks of this technique are ischemia necrosis easily occurred for isolated cartilage and required additional surgery for the donor sit. The patient with scar constitution or undertaken radiotherapy is contraindications. All the managements, appropriate selection of patients, diligent postoperative care, prevented wound infection and using T-tube properly, may increase the rate of successful decannulation. PJP]. PMID- 12772463 TI - [The management of stomal recurrence after laryngectomy]. AB - OBJECTIVE: To study effective prevention and treatment of stoma recurrence after laryngectomy. METHODS: A retrospective review of 23 patients with stomal recurrence after laryngectomy was conducted. Among all 23 cases, 20 cases were referred to Cancer Hospital of Chinese Academy of Medical Science from other hospitals. The etiopathology of stomal recurrence after laryngectomy, feasibilities of various kinds of operative treatment and outcomes of clinical management in this series were analyzed. RESULTS: The subglottic extension and invasion of paraglottic portion of laryngeal cancer and peritreacheal metastasis may result in stomal recurrence. The survival period of 18 cases surgically treated was longer than that of 5 cases untreated or irradiated and/or chemotherapied. The 1- and 5-year survival rates were 40.7% and 20.4%, respectively. The common complication was pharyngeal fistula. Surgical treatment may relieve local pain, improve breathing, resume swallowing and decrease cancer ulcer bleeding. CONCLUSIONS: The overall prognosis of peristomal recurrence after laryngectomy was awful and dismal. Prevention should be stressed in surgical treatment of the subglottic extension, invasion of paraglottic portion and peritreacheal node metastasis. A proper surgical treatment in some cases of stomal recurrence would prolong the lifetime and advance the life quality of the patients. PMID- 12772464 TI - [Preservation of laryngeal and glossal function in surgical treatment of oropharyngeal glossal epiglottic carcinoma]. AB - OBJECTIVE: To study the feasibility, surgical techniques and results of surgical treatment of oropharyngeal glossal epiglottic carcinoma with laryngeal and glossal functions preserved. METHODS: A retrospective review of 21 (17 males, 4 females, age range 44 to 72 years) cases with carcinoma extending from the oropharynx, glossa to the epiglottis treated during 1990 to 2000 was conducted. The clinical stage was T2N0 in 4, T2N1 in 5 and T3N1 in 12 according to 1992 UICC. All the cases were undergone partial resection of the glossal base epiglottis and suproglottic horizontal partial laryngectomy. Seventeen cases received selective neck dissections and all the patients received postoperative radiotherapy (50-60 Gy). RESULTS: The 3-year and 5-year survival rates in this series were 73.3% and 57.1%, respectively. Decannulation and complete preservation of laryngeal and glossal functions were achieved in 20 cases (95.7%), while partial preservation (speaking and deglutition) in one case (4.2%). CONCLUSION: Preservation of the laryngeal and glossal functions in surgical treatment of oropharyngeal glossal epiglottic carcinoma is feasible. Subhyoid approach was proved to be the best way for tumor resection in such an extent. PMID- 12772465 TI - Practice-based interprofessional learning. PMID- 12772466 TI - Lessons learned from implementing multidisciplinary health professions educational models in community settings. AB - This article addresses sustainability elements for multidisciplinary health professions education in communities. The results are based on a 5 year program involving multiple health professions education institutions in seven states within the USA. We offer observations and suggestions to others who plan and implement multidisciplinary programs for health professions educators. Our findings are based on the outcomes of the Community Partnerships in Health Professions Education (CPHPE) initiative funded by the W. K. Kellogg Foundation. Data sources included pre and post surveys of students and faculty, site visits, published evaluations and site reports, and a 2 year follow-up evaluation after the external funding phase. Elements of the partnership that facilitated the sustainability of the models were: leadership, complementary missions, curriculum that mirrors clinical community practice, change from outside in, partnership boards, and faculty development. Those elements that were barriers to sustaining the efforts were: professional identities and territorial boundaries, structural differences, costs, and unclear goals. The multidisciplinary components of the community partnership initiative were the most challenging. However, in most programs, they did succeed and are currently in place. By paying attention to the elements that enhance and detract from teaming, one can plan for more successful multidisciplinary education. PMID- 12772467 TI - Interprofessional perceptions of health care students. AB - Effective interprofessional working, which is widely considered as essential to high-quality health care, is influenced by the attitudes of health care professionals towards their own and other professional groups. Relatively little is known, however, about interprofessional attitudes, particularly of students in health care professions. This study aimed to increase our understanding of students' attitudes towards their own and other professional groups on entry to a programme of professional education. Hypothesised relationships between stereotypes, professional identity and readiness for professional learning were tested by means of a questionnaire survey of 933 undergraduate health care students drawn from five health care groups (medicine, nursing, dietetics, pharmacy and physiotherapy) within a multi-faculty UK university. Positive statistically significant correlations were found between stereotypes, professional identity and readiness for interprofessional learning. As predicted, students identified strongly with their own professional group at the start of pre-registration education. They were also willing to engage in interprofessional learning. More unexpected was the positive association found between heterostereotype and professional identity scores. There are potential benefits of introducing active interprofessional education activities at an early stage of professional preparation to capitalise on students' positive attitudes towards their own and other professional groups. PMID- 12772468 TI - Initial experiences of interprofessional problem-based learning: a comparison of male and female students' views. AB - Few studies have considered the contribution of problem-based learning (PBL) to interprofessional education, and even fewer have examined whether women and men evaluate PBL differently. This paper examines first year occupational therapy and physiotherapy students' evaluations of their initial participation in PBL, during an interprofessional module focusing on communication skills and patient-focused approaches to care. Questionnaire data included attitude ratings and qualitative evaluations of PBL. A total of 133 females and 24 males responded (comprising 83% of the total cohort). Most students were positive that PBL contributed to both personal learning and team-working skills. Both female and male students felt able to express their opinions within the seminar groups and were positive that their understanding of therapists' roles within the multidisciplinary team had increased. However, women expressed rather more trust in the information provided by other students, confirmed greater enjoyment in taking responsibility for their own learning and had more positive views about working with students from another course. In their qualitative comments, more women made reference to enjoying the social aspects of PBL (such as group work, support and collaboration). The gender differences were not substantial but those that were observed support previous researchers' arguments that women are more inclined to be 'connected learners' who value the social aspects of learning contexts. The findings overall suggested that PBL made a positive, well-received contribution to learning during an interprofessional module. PMID- 12772469 TI - Using co-teaching as a means of facilitating interprofessional collaboration in health and social care. AB - In this paper we report the findings of a collaborative enquiry on our experience as tutors co-teaching interprofessional collaboration to a multidisciplinary group of undergraduates. We have different professional/academic backgrounds and the student group included health and social work professionals alongside a number of non-professionals. Our data included our perceptions of the co-teaching experience collected by means of our reflective diaries and reflective conversations during planning and after teaching sessions. We also collected data on student perceptions elicited by means of student evaluations and a student focus group discussion. The data illuminate the process of using co-teaching to role model shared learning and collaborative working within the classroom and highlight the importance of carefully planning co-teaching interaction, including the use of humour, tension, different knowledge bases and styles of debate. The deliberate use of the interactions made possible by coteaching enabled us to create an active learning environment that facilitated the teaching of collaboration. Drawing on our experience, we discuss the considerable potential of using co-teaching to role model collaborative working for multidisciplinary student groups. PMID- 12772470 TI - Multi-agency training for quality: reflections and recommendations. AB - Clinical governance within the National Health Service (NHS) and best value across local authorities in England have emphasised the need for decisions and actions being demonstrably evidence based. In an attempt to progress these related initiatives and facilitate a closer working relationship between NHS Trusts and local government departments, a multi-agency and interprofessional training event on evidence-based practice (EBP) was organised across three boroughs within Tyne and Wear. While there are advantages to different agencies learning and working in partnership to develop quality systems for health improvement, the reality of introducing EBP concepts and creating an evaluative culture within and across public sector organisations represents a major challenge. This article has adopted a case study approach to describe the key lessons learnt through critically reflecting on the planning and delivery of this training event, and it is hoped that it will be of value to other attempting similar projects in the future. PMID- 12772471 TI - Interagency and interprofessional collaboration in community care: the interdependence of structures and values. AB - This paper considers the problems of interagency and interprofessional collaboration in community care in Great Britain from the combined perspectives of UK and US researchers. The research team drew on empirical and theoretical literature from both countries to construct a framework for analysing inter- and intra-organisational theories of joint working. This analysis, supplemented and supported by local case studies conducted by the researchers, generates a framework recommendation against which the government's initiatives for partnership working in the NHS plan 2000 and subsequently can be critically reviewed. In particular, at a time when structural integration--via Care Trusts- is being seriously considered, they highlight the vital importance of integrated systems of goal setting, authority and multidisciplinary service delivery rather than a narrow focus on structural integration alone. PMID- 12772472 TI - Professional predictions: June Huntington's perspectives on joint working, 20 years on. AB - In 1981, June Huntington published a study of social work and general practice which rapidly became influential. In it she offered a sociological analysis of these two occupations to explain the difficulties and conflicts which had arisen between them and to predict the future course of their relationship. In this paper we revisit Huntington's propositions and assess the extent to which her predictions have been realised in the UK context. We adopt her perspective of the two organisational cultures of general practice and social work to consider her claims, particularly with respect to developments in social work. We then outline her depiction of the likely evolution of interprofessional relationships over the past two decades, considering the generalisability of experiments in collaboration, conflict with the medical profession, tensions within social work itself, and the sense of 'crisis' in relationships between professions. To promote debate with colleagues in different disciplines, this paper ends with a series of questions about the likely impact of care trust formation on social work. PMID- 12772473 TI - A portrait of district nursing: its contribution to primary health care. AB - This paper explores the role and contribution of district nursing within primary health care. Examples of how this service is organised within different health care systems are used to discuss the context-dependent nature and challenges of the work. By drawing on UK policy change, health priorities and recent research into district nursing, the paper concludes that, in attempting to deal with the challenges of health care in the 21st century, the significance and potential of district nursing services are frequently overlooked. It is recommended that a research strategy for district nursing should build on the profession's comprehensive understanding of the needs of families and communities and should aim to provide evidence for practice that will further improve patient and client outcomes. PMID- 12772474 TI - Evaluating the introduction of ward-based medical teams on collaboration and patient care. PMID- 12772475 TI - Employers' responsibilities regarding military reservist employees. PMID- 12772476 TI - The nurse's role in changing health policy related to patient safety. AB - Nurses' roles in shaping health policy in the United States related to patient safety have not been fully expanded. This article explores various patient safety issues and how nurses can become involved to shape health policy in this area. PMID- 12772477 TI - The global challenge of road traffic injuries. Foreword. PMID- 12772478 TI - Introduction: the global challenge of road traffic injuries: can we achieve equity in safety? PMID- 12772479 TI - Road traffic safety and health equity: a call to action. PMID- 12772480 TI - Road traffic crashes: operationalizing equity in the context of health sector reform. PMID- 12772481 TI - Equity dimensions of road traffic injuries in low- and middle-income countries. AB - Globally, poorer population groups bear a disproportionate burden of avoidable morbidity and mortality from road traffic injuries. The distribution of road traffic injuries is generally influenced by socioeconomic factors. Poor countries bear a disproportionate burden of injuries and fatalities, and within countries, poor people account for a disproportionate portion of the ill health due to road traffic injuries. The main source of data for this paper was the road traffic injury database of the WHO World Health Report for 1999 supplemented by the WHO Global Burden of Disease Study 2000 report, and published and unpublished works. Fatality rates for 0-4 and 5-14 year olds in low- and middle-income regions, measured as deaths per 100,000 population, were six times the rates for high income regions, while within low- and middle-income regions the rates varied widely. Within poor countries, poor people--represented by pedestrians, passengers in buses and trucks, and cyclists--suffer a higher burden of morbidity and mortality from traffic injuries. In rich countries, children from poor socioeconomic classes suffer more injuries and deaths from road crashes than their counterparts from high-income groups. The disproportionate burden of morbidity and mortality in low- and middle-income countries, and among low socioeconomic groups in those countries, illustrates problems of global inequities in health. The problems can be addressed through policies that focus on the road safety of vulnerable groups. PMID- 12772482 TI - A contemporary analysis of road traffic crashes, fatalities and injuries in Trinidad and Tobago. AB - Road safety, in particular pedestrian safety, is a problem in Trinidad and Tobago. Data were derived from the database of the Traffic and Highway Patrol Unit of the Trinidad and Tobago Police Service. Road traffic crashes in Trinidad and Tobago are largely an urban problem. Four urban areas accounted for nearly three-quarters of reported road traffic crashes, fatalities and injuries. Pedestrians, passengers and drivers accounted for 93% of fatalities and 95% of injuries due to road traffic crashes in 2000. Pedestrians alone accounted for 42% of fatalities and 34% of injuries in 2000. Trends over time show that there has been a decline in fatality rates from 17 deaths per 100,000 population in 1960 to 10 deaths per 100,000 population in 2000, despite rapid motorization. Motorization increased four-fold from 63 registered vehicles per 1000 population in 1960 to 250 vehicles per 1000 population in 2000. In conclusion, effort should be intensified to ensure safety for all road users and, in particular, pedestrians, passengers and drivers. Improved data collection and operational research would improve monitoring and evaluation of policy interventions. PMID- 12772483 TI - Road traffic injuries in Colombia. AB - Road traffic injuries are a leading public health problem in Colombia. Pedestrians are the most vulnerable road users, especially in the main urban centers of Bogota, Medellin and Cali. Data analyzed in this report include official statistics from the National Police and the National Institute of Legal Medicine and Forensic Sciences for 1996-2000, and results of a study conducted at the National University of Colombia in 2000. Methods from the Highway Capacity Manual were used for determining physical and technical variables, and a Geographical Information System tool was used for the location and spatial analysis of the road traffic crashes. Pedestrians accounted for close to 32% of injuries and 40% of the deaths from road traffic crashes. The problem of road traffic crashes existed predominately in urban areas. In the main urban centers, pedestrians constituted nearly 68% of road traffic crash victims. The high level of risky road use behaviors demonstrated by pedestrians and drivers, and inadequate infrastructure for safe mobility of pedestrians in some sections of the road network were the main contributing factors. Major improvements were achieved in Bogota following enhancements to the municipal transport system and other policies introduced since 1995. In conclusion, policies and programs for improving road safety, in particular pedestrian safety, and strengthening urban planning are top priority. PMID- 12772484 TI - Pedestrian traffic injuries in Mexico: a country update. AB - Road traffic injuries in general and pedestrian injuries in particular are a major public health problem in Mexico, especially in large urban areas. Analysis of mortality and road crashes at the national level was done using routine data recorded on death certificates. Fatality rates for different age groups were estimated by region for the year 2000. These data were supplemented by a cross sectional study of pedestrian injuries in Mexico City based on death certificates information for pedestrians who lived and died in Mexico City between 1994 and 1997. Participant observation of physical spaces where crashes occurred was carried out. The spaces were filmed and in-depth interviews of survivors conducted. Road traffic crashes were responsible for approximately 17,500 deaths in Mexico during 2000. The mean age of the victims was 37 years. Mexico lost an average of 30 years of productive life for each individual who died in a traffic crash--525,000 years in 2000. An estimated 9500 (54.3%) of all fatalities were pedestrians, and for every pedestrian death there were 13 others who sustained nonfatal injuries requiring medical care. The overall crude mortality rate for pedestrian injuries in Mexico City was 7.14 per 100,000 (CI 6.85-7.42). A concentration of deaths was observed in 10 neighborhoods at specific types of street environments. The underlying factors included dangerous crossings and the absence or inadequacy of pedestrian bridges, as well as negative perceptions of road safety by pedestrians. In conclusion, this study demonstrates the importance of elucidating the underlying contextual determinants of pedestrian injuries. PMID- 12772485 TI - Strengthening care for injured persons in less developed countries: a case study of Ghana and Mexico. AB - In all countries, the priority for reducing road traffic injuries should be prevention. Nonetheless, there are low-cost ways to strengthen the care of injured persons, that will help to lower the toll from road traffic. The purpose of this review was to elucidate ways to accomplish this goal in the context of less developed countries. Studies selected for this review were obtained by Medline review, selecting on key words such as trauma, injury, trauma care, essential health services, and developing country. Articles pertaining to any country and all available years were considered. In addition, the authors utilized articles from the gray literature and journals from Mexico and Ghana that are not Medline referenced. Studies surveyed point to road safety and other forms of injury prevention, as well as prehospital care, as likely priorities for developing countries. Nonetheless, hospital-based improvements can contribute to decreases in mortality and, especially, decreases in disability. For both prehospital and hospital based care, studies revealed several critical weak points to address in: (1) human resources (staffing and training); (2) physical resources (equipment, supplies, and infrastructure); and (3) administration and organization. The 'essential services' approach, which has contributed to progress in a variety of fields of international health, needs to be developed for the care of the injured. This would define the trauma treatment services that could realistically be made available to virtually every injured person. It would then address the inputs of human resources, physical resources, and administration necessary to assure these services optimally in the different geographic and socioeconomic environments worldwide. Finally, it would identify and target deficiencies in these inputs that need to be strengthened. PMID- 12772486 TI - Road traffic injuries in Kenya: magnitude, causes and status of intervention. AB - Road traffic crashes exert a huge burden on Kenya's economy and health care services. Current interventions are sporadic, uncoordinated and ineffective. This report offers a descriptive analysis of secondary data obtained from a variety of published literature and unpublished reports. Over three thousand people are killed annually on Kenyan roads. A four-fold increase in road fatalities has been experienced over the last 30 years. More than 75% of road traffic casualties are economically productive young adults. Pedestrians and passengers are the most vulnerable; they account for 80% of the deaths. Buses and matatus are the vehicles most frequently involved in fatal crashes. Characteristics of crashes vary considerably between urban and rural settings: pedestrians are more likely to be killed in urban areas, whereas passengers are the majority killed on intercity highways that transverse rural settings. Road safety interventions have not made any measurable impact in reducing the numbers, rates and consequences of road crashes. Despite the marked increase in road crashes in Kenya, little effort has been made to develop and implement effective interventions. Impediments to road traffic injury prevention and control include ineffective coordination, inadequate resources and qualified personnel, and limited capacity to implement and monitor interventions. There is need to improve the collection and availability of accurate data to help in recognising traffic injury as a priority public health problem, raising awareness of policymakers on existing effective countermeasures and mobilizing resources for implementation. Establishment of an effective lead agency and development of stakeholder coalitions to address the problem are desirable. PMID- 12772487 TI - Road traffic injuries in Mozambique. AB - Road traffic injuries affect the economy, health and quality of life of the people of Mozambique. Current road safety programmes are inadequate and inefficient given the magnitude of the problem. Data reported on road traffic crashes in the period 1990 to 2000 from the National Institute for Road Safety, the traffic police and the Central Hospital of Maputo were reviewed. The burden of road traffic injuries in Mozambique is rising, with at least three people killed daily. The age group most affected is 25-38 (39.35%), followed by 16-24 (20.79%). The main causes of crashes include reckless driving, drunken driving, roads with potholes, inadequate signs, lack of protection for pedestrians, and inadequate traffic law enforcement. However, the data are not adequate to reveal the true magnitude of the problem. Data collected by different sources are incomplete and not coordinated with other sources and databases. In urban areas, however, better response to crashes, treatment of the injured, reporting and data collection is attributable to a greater concentration of police and medical facilities. Road traffic safety programmes in Mozambique are inadequate and inefficient, starting with the data collection system. Improvement of injury surveillance systems is needed to help make road traffic safety a national development agenda priority and for developing and implementing road safety policies. For road safety programmes to be effective, government must facilitate stakeholders' involvement, and the clear definition of government activities, civil society activities and public-private partnerships need to be established. PMID- 12772488 TI - Pattern of road traffic injuries in Ghana: implications for control. AB - Road traffic injuries and fatalities are increasing in Ghana. Police-collected crash and injury data for the period 1994-1998 were aggregated and analyzed using the MAAP5 accident analysis package developed by the Transport Research Laboratory, U.K. Published results of recent transport-related epidemiological and other surveys provided an additional data source. According to the 1994-1998 police data, road traffic crashes were a leading cause of death and injuries in Ghana. The other leading causes of death and injuries are occupational injuries which involve non-mechanized farming and tribal conflicts. The majority of road traffic fatalities (61.2%) and injuries (52.3%) occurred on roads in rural areas. About 58% more people died on roads in the rural areas than in urban areas, and generally more severe crashes occurred on rural roads compared with urban areas. Pedestrians accounted for 46.2% of all road traffic fatalities. The majority of these (66.8%) occurred in urban areas. The second leading population of road users affected was riders in passenger-ferrying buses, minibuses and trucks. The majority of these (42.8%) were killed on roads that pass through rural areas. Pedestrian casualties were overrepresented (nearly 90%) in five regions located in the southern half of the country. Efforts to tackle pedestrian safety should focus on the five regions of the country where most pedestrian fatalities occur in urban areas. Policies are also needed to protect passengers in commercially operated passenger-ferrying buses, minibuses and trucks because these vehicles carry a higher risk of being involved in fatal crashes. PMID- 12772489 TI - Speed control in developing countries: issues, challenges and opportunities in reducing road traffic injuries. AB - Speed has been determined to be one of the most common contributing factors in vehicle crashes. This study explores vehicle speed as a factor in the causation of road traffic crashes, using the example of Ghana. It examines the effectiveness of various speed control measures, based on police-reported traffic crashes in Ghana and published works on speed control measures in both industrialized and developing countries. In Ghana, pedestrians were the main victims of road traffic injuries. The dominant driver error assigned by traffic police was loss of control, with the underlying factor being excessive vehicle speeds. The 'speed factor' alone accounted for more than 50% of all Ghanaian road traffic crashes between 1998 and 2000. While the enforcement of speed limits by traffic police may not be affordable for most developing countries, rumble strips and speed humps were found to be effective on Ghanaian roads. Rumble strips installed on the main Accra-Kumasi highway reduced crashes by about 35% and fatalities by about 55%. Reducing vehicle speeds may be one of the most effective interventions to stem traffic crashes in low-income countries. However, setting lower speed limits is not an effective intervention without the traffic law enforcement resources to ensure that limits are followed. Developing countries must also look to other speed reduction measures such as speed bumps and rumble strips, roads that segregate high- and low-speed users, and technological solutions such as speed governors, as well as greater public awareness of the problem. PMID- 12772490 TI - Trends in road traffic crashes and associated injury and fatality in the People's Republic of China, 1951-1999. AB - The burden of road traffic injuries in the People's Republic of China is increasing as evidenced by trends since 1951. Data from the National Statistical Office, Ministry of Communications and the Traffic Administration Bureau were analyzed. Absolute numbers of crashes, fatalities, and injuries, as well as fatalities per 100,000 population and motorization (number of vehicles per 1000 population) were used as indices to measure trends. Regional variations in trends and the characteristics of people injured or killed were also analyzed. Road traffic crashes increased 68-fold, from around 6000 in 1951 to 413,000 in 1999. Excessive speed was the main reported cause of the crashes. The injuries increased 56-fold--from around 5000 to 286,000--and fatalities 97-fold--from 852 to around 84,000--over the same period. The crash, fatality and injury rates also increased after 1985, due to increased motorization spurred by rapid economic growth. The number of four-wheel motor vehicles increased from 60,000 in 1951 to just under a million four-wheel motor vehicles in 1975 and to 10 million in 1987. The number of four-wheel motor vehicles then rose to 50 million in 1999, with an additional 30 million motorcycles. The increase in motorization and fatalities affected all the provinces. Road traffic injuries are the leading cause of death for populations up to the age of 45 years and the leading cause of working-life years lost in China. PMID- 12772491 TI - Road traffic accidents and policy interventions in Korea. AB - The high rate of road traffic crashes, in conjunction with the absence of order on the road, has long been considered a critical social problem in Korea. The Korean public seems to agree that high priority ought to be placed on policies for improving road safety. Using data from government sources, this study describes what has happened in the area of road traffic crashes since 1970, the causes of traffic crashes, and the relative importance of traffic injuries as a cause of death in Korea. Road traffic crashes in Korea increased nearly eight fold, from 37,000 in 1970 to 290,481 in 2000. The fatalities increased three-fold and injuries ten-fold over the same period. Road traffic injuries were the leading cause of death for people under 29. However, through multiple policy interventions, partly in response to the 2002 FIFA World Cup, about two thousand road traffic deaths and nine thousand traffic-related disabilities were averted in 2001 alone. The policy interventions included enforcement of penalties for seven risky driving behaviours, including drunk driving and speeding, installation of traffic-monitoring cameras, financial rewards for citizens who reported traffic violations, introduction of a road safety evaluation system, correction of accident black spots in existing roads, and road safety education programs. Through multiple policy interventions, road traffic crashes in Korea were reduced in a relatively short time period, along with their associated injuries and fatalities. However, road traffic crashes still pose a major public health problem, threatening the quality of life of the Korean people. PMID- 12772492 TI - Road traffic injuries in Thailand: trends, selected underlying determinants and status of intervention. AB - Injuries and deaths from traffic crashes have become a major public health and socio-economic problem in Thailand. Injuries, fatalities and economic losses due to traffic crashes have increased with the rising level of motorization. This study analyzes hospital-based data compiled by the Ministry of Public Health, data compiled by the National Police Office and data compiled by the traffic engineering division of the Department of Highways, Ministry of Transport and Communications. Analysis reveals that 70% of the people injured or killed in traffic crashes are aged 10-39. Men are at four to five times higher risk of death and injury due to traffic crashes than women. The number and rate of traffic injury in Thailand swung from a record low during the economic recovery in the 1980's to record a high during the bubble economy, then declined with the economic crisis in 1997. The economic costs were estimated at U.S.$1.6 billion in 1995. An urban-rural difference in traffic injuries has been recorded with a higher rural case-fatality rate. A number of known behavioral risk factors have been identified, i.e., drunk driving, speeding, substance abuse and failure to use helmets and seat belts. However, determinants of behavior need further investigation. Hazardous road locations have also been mapped. Trends of traffic injuries seem to follow trends of economic growth. Without effective policy and implementation programs to control the determinants, it is expected that traffic injuries will increase as the country recovers from economic crisis. A major pitfall to many current government programs is that they incorporate no systematic evaluation. The fragmented structure of road safety authorities further complicates collaboration and coordination. A broad coalition of stakeholders is needed to catalyze policy action. PMID- 12772493 TI - Injury surveillance. PMID- 12772494 TI - Traffic-related injury prevention interventions for low-income countries. AB - Traffic-related injuries have become a major public health concern worldwide. However, unlike developed or high-income countries (HICs), many developing or low income countries (LICs) have made very little progress towards addressing this problem. Lack of the progress in LICs is attributable, in part, to their economic situation in terms of their governments' lack of resources to invest in traffic safety, cultural beliefs regarding the fatalism of injuries, competing health problems particularly with the emergence of HIV/AIDS, distinctive traffic mixes comprising a substantial number of vulnerable road users for whom less research has been done, low literacy rates precluding motorists to read and understand road signs, and peculiar political situations occasionally predominated by dictatorship and non-democratic governments. How then can LICs tackle the challenge of traffic safety from the experiences of HICs without reinventing the wheel? This paper reviews selected interventions and strategies that have been developed to counter traffic-related injuries in HICs in terms of their effectiveness and their applicability to LICs. Proven and promising interventions or strategies such as seat belt and helmet use, legislation and enforcement of seat belt use, sidewalks, roadway barriers, selected traffic-calming designs (e.g., speed ramps/bumps), pedestrian crossing signs combined with clearly marked crosswalks, and public education and behavior modification targeted at motorists are all feasible and useable in LICs as evidenced by data from many LICs. While numerous traffic-related injury policy interventions and strategies developed largely in HICs are potentially transferable to LICs, it is important to consider country-specific factors such as costs, feasibility, sustainability, and barriers, all of which must be factored into the assessment of effectiveness in specific LIC settings. Almost all interventions and strategies that have been proven effective in HICs will need to be evaluated in LICs and particular attention paid to the effectiveness of enforcement measures. It behooves LIC governments, however, to ensure that only standard, approved safety devices like helmets are imported into their countries. Additionally, LICs may need to improvise and innovate in the traffic safety technology transfer. PMID- 12772495 TI - Afterword: speaking for the victims. PMID- 12772496 TI - Web alert. Oligonucleotide, antibody and peptide therapeutics. PMID- 12772497 TI - Recent advances in immunostimulatory CpG oligonucleotides. AB - The vertebrate immune system has evolved a mechanism to detect CpG motifs within microbial DNA (CpG DNA). Synthetic oligonucleotides containing CpG motifs (CpG ODNs) are potent immunomodulatory molecules and outstanding vaccine adjuvants. A number of recent findings have greatly improved our understanding of the biology of CpG DNA, and the immunological effects of CpG DNA are now recognized to be distinct in mouse and human. The plasmacytoid dendritic cell (PDC) was identified to play a pivotal role in mediating CpG-induced immune responses. So far, the B cell is the only other immune cell subset in humans besides the PDC, equipped with the toll-like receptor-9 to detect CpG motifs. The information on these two prime CpG-sensitive cells has allowed the identification of novel CpG ODNs with distinct functional activity. Together with exciting contributions from animal studies, the way seems to be paved for the successful clinical development of this novel class of molecular therapeutics. PMID- 12772498 TI - Development of novel decoy oligonucleotides: advantages of circular dumb-bell decoy. AB - The inhibition of specific transcription regulatory proteins is a novel approach to regulate gene expression. The transcriptional activities of DNA binding proteins can be inhibited by the use of double-stranded oligonucleotides (ODNs) that compete for binding to their specific target sequences in promoters and enhancers. Transfection of this cis-element double-stranded ODN, referred to as decoy ODN, has been reported to be a powerful tool that provides a new class of anti-gene strategies to gene therapy and permits examination of specific gene regulation. We have demonstrated the usefulness of this decoy ODN strategy in animal models of restenosis, myocardial infarction, glomerulonephritis and rheumatoid arthritis. However, one of the major limitations of decoy ODN technology is the rapid degradation of phosphodiester ODNs by intracellular nucleases. To date, several different types of double-stranded decoy ODNs have been developed to overcome this issue. Circular dumb-bell (CD) double-stranded decoy ODNs that were developed to resolve this issue have attracted a high level of interest. In this review, the applications of decoy ODN strategy and the advantages of modified CD double-stranded decoy ODNs will be discussed. PMID- 12772499 TI - RNA interference: silencing in the cytoplasm and nucleus. AB - Although the discovery that double-stranded RNA is able to silence gene expression was only made five years ago, methods for experimentally silencing genes have already been extended into a broad diversity of organisms, including human cells. RNA interference has also been discovered to function in physiological gene silencing. RNA interference works by causing degradation of targeted mRNAs in the cytoplasm. However, recent results suggest that RNA interference may also silence gene activity in the nucleus by remodeling chromatin and repressing the transcription of targeted genes. PMID- 12772500 TI - Antisense oligonucleotide therapy in cancer. AB - The sequencing of the human genome has highlighted some of the genes that are of importance in disease states. This has provided opportunities for the development of new therapeutics to target a wide range of human diseases. These new drugs are intended to be highly specific; antisense oligonucleotides (ONs) are one such class of new drugs. ONs are short pieces of DNA which hybridize to a specific target mRNA blocking its translation to protein, thereby inhibiting the action of the gene. Several genes known to be of importance in the regulation of apoptosis, cell growth, metastasis and angiogenesis provide a tantalizing prospect for the development of anticancer agents. The phosphorothioate antisense ONs are the current choice for antisense therapy. This article reviews the current strategies for antisense targets in cancer therapy. PMID- 12772502 TI - Cellular uptake and intracellular fate of antisense oligonucleotides. AB - Antisense oligonucleotides and short interfering RNAs are routinely used for gene function analysis and are being developed for clinical applications. The mechanism underlying internalization of free oligonucleotides into cells is poorly understood and inefficient in most cases. Antisense oligonucleotide delivery into ex vivo cells is routinely improved by the addition of cationic lipids. New chemical modifications and vectors allowing improved cellular delivery in vivo are being developed. PMID- 12772501 TI - Peptide-assisted delivery of steric-blocking antisense oligomers. AB - Cationic transport peptides conjugated to steric blocking antisense oligomers (oligos) increase oligo uptake in eukaryotic cell lines, bacteria and mice. Recent reports of arginine-rich transport peptide conjugates strongly suggest that the mechanism of uptake is primarily endocytotic and that previous assay techniques produced confounding artifacts that led to the old non-endocytotic, membrane-penetrating peptide model. The artifacts result from fixing cells for fluorescent microscopy and from using non-trypsinized cells for flow cytometry. Fixing cells redistributes the peptide or peptide-oligo conjugates associated with the outside of cell membranes and trapped in endosomes, giving apparent diffuse cytosolic and nuclear fluorescence. Cationic peptides bound to the outer surface of cells, if not removed, skew fluorescence data obtained by flow cytometry, leading to the earlier conclusions. Upregulation assays now provide a tool for comparing the efficacy of conjugates, measuring oligo uptake by quantitating antisense activity of conjugates. These assays, developed in cell culture and mouse models, are faster and have higher signal-to-noise ratios than downregulation assays. Thus, a convenient and effective method now exists to screen transport peptides. PMID- 12772503 TI - Therapeutic recombinant proteins: trends in US approvals 1982 to 2002. AB - Recombinant proteins have become an important class of therapeutics since the first product was approved for marketing in the US in 1982. To determine trends in the clinical development and approval processes for therapeutic recombinant proteins, data for 270 products that entered clinical study in the period 1980 to 2002 were collected and analyzed. Mean clinical and approval phase lengths for 54 US-approved products stratified by time period, therapeutic indication, protein type, review rating and FDA review center are compared. Approval success rates, calculated using three methods that differ only in the definition of success, and the variation of approval success rates over five time periods are presented. PMID- 12772504 TI - Therapeutic antibodies against viral hepatitis. AB - Antibodies have the potential to be immunotherapeutic agents, used either as stand-alone therapy or as an adjunct for managing chronic viral infection. In addition, antibodies may be used prophylactically in individuals who have been accidentally exposed to hepatitis B virus (HBV) or hepatitis C virus (HCV), or to prevent re-infection of the liver in patients who have undergone liver transplantation. Human monoclonal antibodies to HBV and HCV were generated and their ability to reduce viral load was tested in different animal model systems, the Trimera mouse model and HBV-carrier chimpanzees. These antibodies were further developed and are currently being studied in clinical trials for chronic HBV or HCV and in liver transplant patients. The antibodies were shown to be safe, tolerable and could significantly reduce viral load. Their ability to inhibit HCV re-infection in the transplanted liver is being evaluated. PMID- 12772505 TI - Monoclonal antibodies to two epitopes of beta-human chorionic gonadotropin for the treatment of cancer. AB - Human chorionic gonadotropin (hCG) is expressed by many histological types of cancer and may play an important role in tumor maintenance and progression. Vaccination of patients with the therapeutic peptide Avicine (CTP37; AVI BioPharma Inc/SuperGen Inc), that contains 37 amino acids from the carboxyl terminus (CTP37; AVI BioPharma Inc/SuperGen Inc) of hCG, can result in two distinct antibody responses to separate epitopes within the peptide. Colorectal cancer patients who develop both anti-hCG responses show a significant improvement in median survival time. These observations provide a compelling rationale for the development of two human monoclonal antibodies (mAbs), one for each of the epitopes within the 37 amino acid peptide region of hCG. Two such human mAbs, both exhibiting a high degree of specificity and affinity have been prepared using XenoMouse technology. These mAbs may prove useful in multiple clinical settings for the treatment of various cancers. Treatment options may include passive immunotherapy with both mAbs, mixed passive supplement to active specific immunotherapy with Avicine and conjugation of the mAbs with radioisotopes or cytotoxic drugs. The requirement for dual mAb therapy is consistent with current trends in the development of complex, non-toxic therapies for cancer. PMID- 12772507 TI - The immunogenicity of therapeutic cytokines. AB - Therapeutic cytokines that modulate immune responses are designed to enhance host defense to combat tumors and chronic infections. In general, cytokines are pleiotropic molecules and mediate both systemic and local immune activities. Therapeutic recombinant human cytokines currently in clinical use include interferons (IFN alpha, IFN beta and IFN gamma), interleukins (IL-2 and IL-12) and hematopoietic factors such as granulocyte colony-stimulating factor, granulocyte-macrophage colony-stimulating factor, erythropoietin and thrombopoietin. Their use as therapeutic agents has been challenging since the safety and efficacy of these products are complicated by immunogenicity issues. PMID- 12772506 TI - Selective gene regulation with designed transcription factors: implications for therapy. AB - The prospect of selectively regulating gene expression is highly appealing, both for laboratory investigations and for potential therapeutic applications. This possibility has recently become a reality through the use of designed transcription factors (DTFs). This approach makes use of the DNA-binding domains of native transcription factors as scaffolds, but then introduces new DNA recognition capabilities that allow selective recognition of the regulatory regions of individual genes. The DTF strategy allows either upregulation or downregulation of transcription. This differs from approaches such as antisense or RNA interference that are intended only to inhibit gene expression. Much of the work to date on DTFs has exploited the modular nature of Cys2-His2 zinc finger (Zif) modules that, when coupled with the use of phage display or other combinatorial library techniques, allow the creation of novel and selective DNA recognition domains. These domains can then be linked to transcriptional regulatory moieties, including transactivator or repressor domains, to modulate gene expression. Recent progress has demonstrated that DTFs can selectively regulate the transcription of endogenous genes in mammalian cells and in living animals. PMID- 12772508 TI - Immunogenicity of therapeutic monoclonal antibodies. AB - There are currently 13 monoclonal antibodies or antibody constructs approved for therapeutic use in the US and at least another 400 products are in clinical testing or undergoing regulatory review. Despite widespread use and development of therapeutic monoclonal antibodies, limited information exists regarding the incidence and consequence of immune antibodies generated against many of these products. In this review, analytical methodology and recent data on the immunogenicity of approved therapeutic monoclonal antibodies will be presented. The implications of these findings in relation to potential concerns for repeated or chronic treatment with these powerful and specific therapeutic agents will be discussed. PMID- 12772509 TI - Enhancing immunogenicity by CpG DNA. AB - Bacterial DNA and oligonucleotides containing unmethylated CpG dinucleotides (CpG DNA) can stimulate immune responses and have potential for use as novel agents to enhance immunogenicity. CpG DNA can interact with toll-like receptor 9 and cause activation through a myeloid differentiation primary response gene (MyD88) dependent signaling pathway. Due to its pattern of immune cell activation, CpG DNA can induce a cytokine milieu to promote T-helper cell responses and serve as an adjuvant. Furthermore, CpG DNA can provide protection against pathogens in animal models and has therapeutic applications in clinical settings such as in cancer and allergy. PMID- 12772510 TI - Technology evaluation: reticulose, advanced viral research. AB - Advanced Viral Research Corp is developing reticulose, a nontoxic peptide nucleic acid preparation, for the potential treatment of viral infection. The compound is undergoing phase II clinical trials for the potential topical treatment of human papillomavirus infection in the US. Reticulose has also been investigated in clinical studies for the treatment of HIV infection in Barbados. Additionally, the drug is being investigated for the potential treatment of adenovirus infection, and is being evaluated in adjuvant arthritis models by the Weizmann Institute of Science. Investigations into the use of reticulose to reduce the toxic effects of cancer treatment are underway in Israel. Phase I trials in patients with cachexia associated with HIV, lymphoma and solid tumor, have also been initiated in Israel. PMID- 12772511 TI - Technology evaluation: epratuzumab, Immunomedics/Amgen. AB - Immunomedics and Amgen are developing the unconjugated form of the anti-CD22 humanized monoclonal antibody, epratuzumab, for the potential treatment of lymphoma, non-Hodgkin's lymphoma (NHL) and certain autoimmune conditions. Immunogenics is also developing the radioisotope-linked (90Y) form of epratuzumab. The monoclonal antibody is currently undergoing phase III clinical trials for the potential treatment of aggressive lymphoma. By January 2003, epratuzumab had received Orphan Drug status in the US for its potential in the treatment of NHL. PMID- 12772512 TI - Technology evaluation: lerdelimumab, Cambridge Antibody Technology. AB - Cambridge Antibody Technology is developing lerdelimumab (CAT-152), the lead in a series of human anti-TGF beta 2 antibodies, for its potential in preventing post operative scarring in patients undergoing surgery for glaucoma (trabulectomy). It also has potential for the treatment of fibrosis, cataract, retinopathy and connective tissue disease. PMID- 12772513 TI - Technology evaluation: TriGem, Titan. AB - Titan (formerly Trilex) is developing TriGem, a monoclonal antibody directed against I17, for the potential treatment of advanced cancers that express the GD2 ganglioside, including melanoma, small-cell lung cancer, neuroblastoma and sarcoma. Phase II trials of TriGem for use in advanced melanoma were ongoing by June 2002. TriGem also has potential for the treatment of psoriasis, for which phase II trials had begun by 1996. PMID- 12772514 TI - Recent advances in the total management of colorectal cancer. PMID- 12772515 TI - Surgical management of colorectal metastases to the liver. AB - Liver metastases develop in 50% to 60% of patients with colorectal carcinoma. Surgical resection offers a good five-year survival rate and the only chance of cure. In this article, the natural history and risk factors for the development of colorectal liver metastases are discussed, followed by the diagnostic work-up towards surgical resection. Various issues pertaining to surgical management, such as patient selection, timing of resection, perioperative mortality and morbidity, prognostic factors, adjuvant and neoadjuvant therapy and repeat resection are reviewed. PMID- 12772516 TI - Flat and depressed lesions of the colon and rectum: Pathogenesis and clinical management. AB - INTRODUCTION: It is not clear whether flat lesions play a role in the pathogenesis of colorectal carcinoma. Flat lesions are being increasingly recognised with new colonoscopic techniques. MATERIALS AND METHODS: A total of 10,939 consecutive colonoscopies were performed over a 9-year period. After bowel preparation with polyethylene glycol electrolyte lavage solution, high-resolution video colonoscopy and indigocarmine spraying were performed to detect flat lesions. All lesions suggesting neoplastic change were removed by polypectomy or surgery. Cancers invading beyond the submucosal layer were excluded from this analysis. The gross appearance of flat-type lesions was classified as flat elevated type or flat depressed type based on the presence or absence of central depression. RESULTS: A total of 5408 neoplastic lesions were index lesions, including 5035 adenomas and 373 carcinomas (124 with submucosal invasion). The prevalence of flat depressed and flat elevated lesions were 2.8% and 18.1%, respectively. Submucosal invasion rates were 17.1% in the flat depressed, 0.8% in the flat elevated, 1.6% in the sessile, 4.0% in pedunculated lesions and 9.3% in creeping lesions. The submucosal invasion rate in the flat depressed lesions was significantly higher than in any others, except for creeping lesions (P = 0.06). The percentage of flat elevated and flat depressed carcinomas among all carcinomas invading the submucosa was 6.5% and 21.0%, respectively. CONCLUSION: Flat lesions were common during routine colonoscopy. One-quarter of colorectal cancers may be derived from flat lesions. Training in dye spray technique may result in a higher detection rate of flat colonic lesions. PMID- 12772517 TI - Total mesorectal excision (TME)--twenty years on. AB - INTRODUCTION: The results of total mesorectal excision (TME) for rectal cancer were first reported 20 years ago. The superior outcome in the original two papers was attributed to the complete excision of the mesorectum, preference for anterior resection over abdominoperineal resection, application of cytotoxic solutions to eradicate exfoliated tumour cells and specialisation in rectal cancer surgery. A review was conducted to examine the evidence from the literature accumulated over the past 20 years in support of this thesis. METHODS: A Medline search of all studies was carried out pertaining to the issues of mesorectal metastases, anterior resection versus abdominoperineal resection, exfoliated tumour cells in rectal cancer and specialisation in rectal cancer surgery. RESULTS: Mesorectal metastases can be found up to 5 cm from the apparent distal edge of a rectal cancer. They occur in pT3 and pT4 tumours. Their presence indicates a poorer prognosis. There is evidence that TME improves outcomes in mid and low-rectal cancers but its role in upper rectal and rectosigmoid cancers is controversial. It has been difficult to demonstrate the superiority of anterior resection over abdominoperineal resection as comparative studies report similar outcomes. Exfoliated tumour cells have been demonstrated to contribute to anastomotic recurrence and this probability may be reduced by the use of cytotoxic agents to irrigate the rectal stump. Multiple studies have shown that specialisation in rectal cancer surgery result in lower postoperative morbidity and mortality, local and distant recurrence rates and higher rates of sphincter saving resections. CONCLUSION: The majority of tenets espoused in the original papers on TME have found support in follow-up studies since their publication. PMID- 12772518 TI - Practical issues in adjuvant therapy for rectal cancer. AB - INTRODUCTION: While the standard adjuvant therapy for rectal cancer includes radiation therapy (RT) and chemotherapy (CT), the optimal treatment combination and sequencing have yet to be determined. In recent years, a large number of clinical trials have been reported in this rapidly evolving field. MATERIALS AND METHODS: A review of pivotal trials in rectal cancer was undertaken with a focus on the important recent literature that has determined the current approach to adjuvant treatment. RESULTS: Multiple well-conducted, randomised studies have demonstrated that, for patients with stage II and III rectal cancer, the combination of adjuvant CT and RT reduces the risk of local recurrence (LR) and distant recurrence, improves overall survival and should form part of the standard treatment. Due to the reduced risk of LR, the absolute benefit from adjuvant RT is less if optimal oncologic surgery has been performed. Theoretical advantages and preliminary clinical results suggest that delivering part of the adjuvant therapy preoperatively will produce superior results. Ongoing randomised trials will define the relative merits of long-course RT (with CT) and short course RT alone as preoperative therapy. Preoperative combined modality therapy might increase the rate of sphincter preservation, although definitive data supporting this is still being awaited, whereas RT alone does not facilitate sphincter preservation. CONCLUSION: Management of patients with rectal cancer is complex and requires ongoing close collaboration between the colorectal surgeon, medical oncologist and radiation therapist in order to achieve the best possible outcomes. The optimal combination of CT and RT will be defined by current clinical trials. PMID- 12772519 TI - Chemoprevention of colorectal cancer--experimental approach and clinical applications. AB - INTRODUCTION: The holy grail of cancer treatment is prevention. Chemoprevention is the use of specific agents to prevent, inhibit or reverse the process of cancer formation before malignancy. This review aims to explore the clinical approaches to investigation for chemoprevention and to discuss the more promising agents that have proceeded on to clinical trials. METHODS: A Medline search was done using the words colorectal cancer, chemoprevention, models, surrogate markers and prevention clinical trials. RESULTS: Research into chemopreventive agents is highly dependent on an effective model in which studies may be performed and from which results may be extrapolated into the human or clinical setting. As carcinogenesis is a prolonged biological event, intermediate surrogate markers that will reliably predict future cancer risk are usually studied. Various models are used with varying impact. Among some of the more recent agents being explored, the COX-2 inhibitors are emerging as strong potential agents. However, there are significant side-effects that need to be overcome first before routine use may be considered. CONCLUSIONS: Although there is presently no agent available for the safe, widespread use for the chemoprevention of colorectal cancer, research into this area is rapidly progressing and may in the future change the paradigm of cancer treatment. PMID- 12772520 TI - Current concepts of tumour metastasis. AB - BACKGROUND: Tumour metastasis remains the principal cause of treatment failure and poor prognosis in patients with cancer. Recent advances in our understanding of the biology of metastasis are providing novel potential targets for anti cancer therapies. AIM: This paper reviews the current concepts in tumour metastasis. METHODS: A review of Medline publications relating to the molecular biology and therapy of human tumour metastasis was conducted. RESULTS AND DISCUSSION: Early metastasis models were based upon the premise of uninterrupted tumour growth, with the inevitable formation of distant metastases and eventual death of the patient. However, current research suggests that metastasis is an inefficient process governed by several rate-limiting steps, and that failure to negotiate these steps can lead to tumour dormancy. Successful metastatic tumour growth depends upon appropriate tumour-host microenvironment interactions and, ultimately, the development of vascularised metastases post-extravasation in the target organ. An understanding of the molecular mechanisms involved in this dynamic process will aid in the identification of therapeutic targets that may allow earlier diagnosis and more specific therapies for patients with metastasis. PMID- 12772521 TI - Current status in imaging of colorectal liver metastases. AB - Colorectal cancer is a common malignancy that often metastasizes to the liver. Evaluation of the liver for metastases is important for staging and prognosis. Resection of limited hepatic metastases can result in prolonged survival. Imaging is vital to select appropriate patients to avoid unnecessary surgery. A variety of modern imaging tools can be used to detect and characterise hepatic lesions. These include contrast enhanced helical computed tomography (CT), CT arterial portography, magnetic resonance imaging (MRI) and intraoperative ultrasound (IOUS). Each has their advantages as well as disadvantages. Besides accuracy, availability, cost and presence of expertise determine the optimal modality or combination of modalities to use. The appearance of metastases on imaging can be correlated with histopathological abnormalities. Though metastases can often be distinguished from benign lesions, their appearances are by no means pathognomonic. Follow-up imaging for interval change and for assessment of response is often required. PMID- 12772522 TI - Computed tomographic appearance of colorectal hepatic metastases. AB - INTRODUCTION: This article reviews the various computed tomography (CT) appearances of hepatic metastases from colorectal primaries and assesses the frequency of occurrence of the various patterns. MATERIALS AND METHODS: This is a retrospective study of the CT appearances of histologically proven colorectal hepatic metastases in a group of 52 patients who had undergone surgical hepatic resection between January 1994 and December 2001. A total of 74 hepatic metastatic lesions were reviewed. All lesions were examined in the portal venous phase. RESULTS: A discernible rim was seen in 54 lesions (73%). Thick rim was present in 36 lesions (48.6%) and thin rim in 18 lesions (24.3%). Enhancement of the rim was present in 62 cases (83.8%). Increased central attenuation was seen in 38 lesions (51.4%). Of these, the centre was heterogeneous in 76.3% and scar like in 23.7%. A non-enhancing rim was seen in 12 lesions (16.2%) which appeared as lesions with "bevelled edge". Thick enhancing rim with non-enhancing centre was the most common combination in 15 lesions (20.3%). CONCLUSION: An enhancing rim could be seen in 83.8% of lesions. Increased central attenuation was present in 51.4% of the lesions. Familiarity with the various CT appearances may facilitate identification and diagnosis of colorectal liver metastases. PMID- 12772523 TI - Hepatic resection for colorectal metastases to the liver: The National Cancer Centre/Singapore General Hospital experience. AB - OBJECTIVE: To review and evaluate trends for short and long-term outcomes of patients who have undergone liver resection for colorectal metastases in a single institution over a period of 7 years. MATERIALS AND METHODS: A retrospective review and analysis of clinicopathologic data and outcome of potentially curative liver resection for colorectal metastases in a series of 96 patients from January 1994 to December 2001 was done. Data were statistically analysed. Perioperative mortality, morbidity, overall survival and disease-free survival rates were reported. RESULTS: A total of 96 patients underwent potentially curative liver resection for colorectal metastases. There were 64 males (66.7%) and 32 females (33.3%) with a median age of 60 years. There was no perioperative mortality. Postoperative morbidity was 7.2%. The median length of follow-up for the entire cohort of post hepatic resection was 29 months with 1-year and 3-year overall survival rates of 99% and 71%, respectively. The disease-free survival rates were 76% and 48%, respectively. Minor resections for the liver metastases showed significantly better overall survival. Primary tumour in the colon compared to rectum, disease-free interval (DFI) < 12 months and adjuvant chemotherapy showed a trend towards shorter overall survival. Carcinoembryonic antigen (CEA) levels greater than 100 ng/mL significantly shortens the disease-free survival. CONCLUSION: Liver resection for colorectal metastases in our institution is a safe and effective treatment option in properly selected patients, which currently represents the best chance for long-term survival and possibly "cure". PMID- 12772524 TI - Neoadjuvant chemotherapy in the management of colorectal metastases: A review of the literature. AB - INTRODUCTION: Five-year survival rates of between 20% and 45% are achievable in patients following hepatectomy for liver metastases from colorectal cancer. Sadly, a great majority of patients present with unresectable liver disease with virtually no long-term survival. This paper reviews the development and role of preoperative chemotherapy in trying to downstage unresectable liver disease. METHODS: The author retrieved most of the citations by performing a PubMed search for studies related to preoperative chemotherapy followed by surgical resection of liver metastases. RESULTS: Larger series have reported resectability rates ranging from 10% to 40% for unresectable colorectal liver metastases after treatment with preoperative chemotherapy. Chemotherapy generally consisted of oxaliplatin added to 5-fluorouracil/leucovorin administered in a chronomodulated fashion. A median of about 5 to 10 months of treatment was necessary for adequate tumour reduction. The 5-year survival rates, after surgery, in completely resected patients were between 30% and 60%. The majority of these patients will relapse, of which half of these will remain confined to the liver. Patients who are able to undergo a repeat hepatectomy may achieve long-term survival rates similar to the initial resection. Preliminary data from a recent study suggest that initial treatment with an oxaliplatin-containing regimen may lead to a higher resectability rate compared to an irinotecan-containing combination (18% versus 7%). Selective intra-arterial infusion has not demonstrated a clear advantage over systemic infusion and, if administered preoperatively, it may even compromise the safety of surgery. CONCLUSIONS: Palliative chemotherapy remains the standard of care for unresectable colorectal liver metastases. However, in carefully selected patients, neoadjuvant chemotherapy may be able to downstage the disease adequately for curative resection. PMID- 12772525 TI - Colorectal hepatic metastases: Role of radiofrequency ablation. AB - INTRODUCTION: Radiofrequency ablation (RFA) is a new minimally invasive treatment that has been increasingly used in the treatment of liver metastases. This review aims to outline the principles governing the use of RFA and to examine its role when applied to the management of colorectal liver metastases. METHODS: A Medline search of experimental and clinical studies relating to the use of RFA in the management of colorectal hepatic metastasis was carried out. RESULTS: RFA is currently used as an alternative to surgery in patients with unresectable disease, and sometimes as its companion, allowing patients hitherto considered unsuitable for resection to become surgical candidates. RFA has been shown to be safe and well tolerated, with few major complications and minimal patient discomfort. Although its use is unlikely to achieve cure in such patients, it has a definite role in palliation and relief of symptoms. Long-term data, when these become available, may also show improved survival. However, because RFA is a local ablative therapy, it does not address the progressive and systemic nature of colorectal carcinoma. CONCLUSIONS: RFA is an important alternative/complimentary tool in the management of colorectal hepatic metastases. Combining RFA with surgery or chemotherapy may reduce the incidence of local and systemic relapse. PMID- 12772526 TI - The molecular biology of peritoneal carcinomatosis from gastrointestinal cancer. AB - INTRODUCTION: Peritoneal carcinomatosis is a frequent form of disease progression in gastrointestinal cancer, and all too often is a preterminal event with a median survival of only 6 months. Despite the introduction of aggressive surgical and chemotherapeutic approaches, any significant improvement in survival is unlikely until we better understand the molecular biology of peritoneal metastasis. METHODS: A Medline search and review of references was undertaken to identify all manuscripts in the English language concerned with peritoneal metastasis from gastrointestinal cancer. RESULTS: Peritoneal carcinomatosis involves a complex sequence of interdependent steps. The injured peritoneum is a rich source of cytokines and growth factors that facilitate tumour proliferation and invasion in the postoperative abdomen. Peritoneal tumour adhesion is dependent on adhesion molecules, such as CD44, and the beta-1 integrins. Invasion of the mesothelium involves, at least in part, a process of tumour-induced mesothelial apoptosis. Matrix metalloproteinases, such MMP-7, facilitate stromal invasion, but the role of other proteases in invasion remains to be elucidated. To date, the significance of angiogenesis in the peritoneal metastatic cascade is unknown. CONCLUSION: The molecular biology of peritoneal carcinomatosis is only just beginning to be understood. Further research into the mediators of the peritoneal metastatic cascade is needed if more effective therapeutic strategies are to be developed for this invariably fatal, yet unfortunately common, condition. PMID- 12772527 TI - 16th Gordon Arthur Ransom Oration. Navigating in the sea of science and technology. PMID- 12772528 TI - Stuart Gan Memorial Lecture 2002. HIV/AIDS in children. AB - The incidence of human immunodeficiency virus (HIV) infection and acquired immunodeficiency syndrome (AIDS) in children has risen steadily over the years and has become an increasing cause of morbidity and mortality during childhood. More than 90% of all children with HIV infection acquired their infection at birth from HIV-infected mothers. With increasing evidence of heterosexual HIV transmission, the number of infected women and consequently their children is increasing. The transmission rate of HIV from mother to infant varies from 20% to 40%. Children can be affected directly through HIV infection and AIDS, and indirectly by the effects of their parents' HIV infection, including being orphaned, discrimination, infant abandonment and negative impact on children's education. Another concern regards children who are most vulnerable to HIV infection and AIDS, such as child prostitutes and children in difficult circumstances. Recommended actions include prevention of parental HIV infection, prevention of mother-to-child HIV transmission, addressing child prostitution, prevention of child labour and improving work conditions, assistance to street children, addressing discrimination, solving the problem of children orphaned by AIDS, reducing HIV-related child abandonment and creating new roles for schools in reducing negative social impact. Although the largest number of HIV-infected persons are from sub-Saharan Africa, the annual incidence of HIV infection in Asia is escalating alarmingly. As reported by the Joint United Nations Programme on HIV/AIDS (UNAIDS) and the World Health Organization in December 2001, HIV infection has already killed more than 20 million people worldwide since the beginning of the epidemic and has infected more than 40 million people; almost half of them were women. There are at least 7 million people living with HIV/AIDS in the Asia-Pacific region. The numbers of HIV-infected women and children are increasing at an alarming rate. Over 1 million children are HIV-infected; in the year 2000 alone, 600,000 children acquired HIV infection. PMID- 12772529 TI - Autoimmune liver disease in children. AB - Autoimmune liver disorders are characterised by an inflammatory liver histology, circulating non-organ specific autoantibodies and increased levels of immunoglobulin G (IgG) in the absence of a known aetiology. They respond to immunosuppressive treatment, which should be instituted as soon as diagnosis is made. Liver disorders with a likely autoimmune pathogenesis include autoimmune hepatitis (AIH) and autoimmune sclerosing cholangitis (ASC). Two types of AIH are recognised according to seropositivity for smooth muscle and/or antinuclear antibody (SMA/ANA, type 1) or liver kidney microsomal antibody (LKM1, type 2). There is a female predominance in both. LKM1-positive patients tend to present more acutely, at a younger age, and commonly have immunoglobulin A (IgA) deficiency, while duration of symptoms before diagnosis, clinical signs, family history of autoimmunity, presence of associated autoimmune disorders, response to treatment and long-term prognosis are similar in both groups. The most common type of paediatric sclerosing cholangitis is ASC. The clinical, biochemical, immunological and histological presentation of ASC is often indistinguishable from that of AIH. In both, there are high IgG, non-organ specific autoantibodies and interface hepatitis. Diagnosis is made by cholangiography. Children with ASC respond to immunosuppression satisfactorily and similarly to AIH in respect to remission and relapse rates, times to normalisation of biochemical parameters and decreased inflammatory activity on follow-up liver biopsies. However, the cholangiopathy can progress and there may be an evolution from AIH to ASC over the years, despite treatment. Whether the juvenile autoimmune form of sclerosing cholangitis and AIH are 2 distinct entities, or different aspects of the same condition, remains to be elucidated. PMID- 12772530 TI - Craniofacial anomalies: Clinical and molecular perspectives. AB - The first three disorders discussed are abnormalities of bone: too little bone in cleidocranial dysplasia caused by mutations in RUNX2; too much bone in fibrodysplasia ossificans progressiva with overexpression of BMP4; and abnormal bone in McCune-Albright syndrome and fibrous dysplasia caused by mutations in GNAS1. Disorders of the sonic hedgehog signaling network are discussed next, including holoprosencephaly and the nevoid basal cell carcinoma syndrome, the former being caused by sonic hedgehog (SHH) mutations and the latter being caused by patched mutations (PTCH). PMID- 12772531 TI - 7th Yahya Cohen Lecture. Significance of the lateral epiphysis of the acetabulum to hip joint stability. AB - Normal development of the acetabulum is crucial to the growth and stability of the hip. Twenty-five New Zealand white rabbits (postweaning) weighing 1.5 to 2.2 kg were used. Both hips were exposed through an anterolateral approach. On the right hip, a fixed area of superolateral physeal cartilage was damaged with drills. Sham open reduction was performed for the left hip. Radiographic changes of the right hips were evident at 6 weeks. At 12 weeks, the right hip dislocated in a posterior and superior direction. The left hip remained normal. Histopathologic analysis correlated strongly with the radiographic findings. There were thinning of cartilage cells of the acetabulum, with disorganisation. The acetabular roof was poorly formed. The lateral acetabular epiphysis is vital to the development of the acetabular roof. Damage to this epiphysis may result in acetabular maldevelopment and subsequent hip instability. We tested the hypothesis that abnormality in this epiphysis can give rise to abnormal acetabular development. PMID- 12772532 TI - Predicting significant hyperbilirubinaemia and early discharge for glucose-6 phosphate dehydrogenase deficient newborns. AB - INTRODUCTION: This study aims to assess the usefulness of day 3 (49 to 72 hours) pre-phototherapy total serum bilirubin (TSB) in predicting subsequent significant hyperbilirubinaemia (SHB) and the feasibility of early discharge for term and near-term glucose-6-phosphate dehydrogenase (G6PD) deficient newborns. MATERIALS AND METHODS: This prospective cohort study involved in born G6PD deficient neonates who were > or = 35 weeks and weighted > or = 2000 g at birth. TSB levels and phototherapy requirements in their first two weeks of life were studied. Day 3 pre-phototherapy TSB in the subgroup weighing > or = 2500 g at birth was analysed for its value in predicting subsequent SHB. RESULTS: Of the 129 neonates, 58 (45%) required phototherapy in the first week. Of these, only 4 patients (3.1%) needed phototherapy to be restarted in the second week. Seventy one (55%) neonates did not require phototherapy at all. In the absence of SHB in the first week, the probability of its development in the second week was zero (95% confidence interval, 0 to 0.057). In the subgroup weighing > or = 2500 g at birth, day 3 pre-phototherapy TSB < or = 154 umol/L predicted no measurable risk of subsequent SHB (sensitivity, 100%; 95% confidence interval, 91.4% to 100%; negative predictive value, 100%; 95% confidence interval, 86.7% to 100%). CONCLUSIONS: G6PD deficient newborns without SHB in their first week of life were at no measurable risk of its development in the second week. Day 3 pre phototherapy TSB in the subgroup weighing > or = 2500 g was useful for predicting the risk of subsequent SHB. Low-risk infants, thus identified, may be eligible for discharge on or before day 7 of life. Evidence-based early discharge can decrease the social and financial burden of G6PD deficiency in Singapore. PMID- 12772533 TI - Runme Shaw Memorial Lecture 2002. Modern management of colorectal polyps: Are they all premalignant? AB - There are various types of polyps in the colon and rectum. Most colorectal polyps in the colon and rectum are either adenomas or hyperplastic polyps. In general, adenomas are considered to be premalignant. Adenoma-carcinoma sequence is an established theory of carcinogenesis in the large bowel in Western countries. However, small adenomas are ubiquitous phenomena, and it is questionable whether they are actually precancerous. Most small adenomas did not become larger in our colonoscopic follow-up study of small adenomas measuring 5 mm or less. It was considered safe to leave such small adenomas behind, particularly in a situation in which follow-up examination is periodically performed, such as in patients who have undergone colorectal cancer surgery before. In Japan, small flat carcinomas have been reported as an important precursor of advanced colorectal carcinoma. They are also found in Western population. The malignancy rate of small flat and depressed lesions was significantly higher than that of polypoid lesions. Although small flat carcinomas are less frequently seen than polypoid-type carcinomas in the early stages, they are considered to be an important precursor of advanced colorectal cancer. PMID- 12772534 TI - [Scarring, a complex phenomenon]. PMID- 12772535 TI - [Wound dressing, the instruments to use]. PMID- 12772536 TI - [Make-up for the correction of medical problems]. PMID- 12772537 TI - [How to recognize hyperactivity syndromes in children]. PMID- 12772538 TI - [Diabetes. The fact of daily injections]. PMID- 12772540 TI - [The diet for the severely burned]. PMID- 12772539 TI - [A person in a precarious situation. When nurses question their practice]. PMID- 12772541 TI - [Meeting with a president "determined to succeed"]. PMID- 12772542 TI - Improving health outcomes in diverse and vulnerable populations: building on the experience of the Centers for Medical Treatment Effectiveness in Diverse Populations (MEDTEP). PMID- 12772543 TI - Establishing centers to address treatment effectiveness in diverse ethnic groups: the MEDTEP experience. AB - OBJECTIVE: To investigate racial, ethnic, and socioeconomic disparities in health outcomes amenable to improvement through more effective delivery of health care services. DESIGN: The Medical Treatment Effectiveness Program (MEDTEP) Research Centers on Minority Populations were a group of centers in the USA funded to improve the effectiveness of medical diagnosis and treatment, to provide technical assistance to ethnic minority health researchers, to train new researchers, and to disseminate information to help ethnic minority patients and their health care providers. RESULTS: Centers often provided many specific findings related to assessment of the magnitude of disparities in health outcomes and to approaches for eliminating these outcomes. The Centers were able to build community partnerships using an approach now defined as community-based participatory research. Centers changed the culture of their institutions by making them more aware of the need to train diverse investigators and do more to eliminate health disparities. CONCLUSION: A key to the success of the Centers has been the unification of a cadre of committed investigators dedicated to the mentoring of minority health researchers and to the elimination of ethnic and socioeconomic disparities in health. The MEDTEP Centers provide a model but there remains a need for continued work. PMID- 12772544 TI - Perceptions of diabetes among patients in an urban health care system. AB - OBJECTIVES: A major problem facing health care providers today is adherence to treatment regimens by patients. Adherence is of even greater significance for patients with diabetes, who shoulder a great deal of responsibility in their disease management. Perceptions of diabetes have been found to play a major role in adherence. The effects of race and socioeconomic status on the disease perceptions remain unclear. This exploratory study encompassed two themes: (1) assessing perceptions of diabetes among African American and white American adults with diabetes who were patients in 1994 in a large, Midwestern, urban health care system and (2) examining the psychometric properties of the measurement instruments used to study perceptions. DESIGN: A stratified random sampling scheme (by race and socioeconomic status (SES)) was used. Diabetes perceptions were measured using three scales from the Meaning of Illness Questionnaire assessing the impact, loss, and stress associated with diabetes. Perceptions of physician efficacy were also measured. The study population consisted of 50 (68% response rate) African American and white American patients aged 18-65 years. RESULTS: No differences in SES were found between the African American and white American participants (p = 0.44). However, the African Americans in the study indicated a greater sense of loss associated with diabetes than the white Americans in the study (p < 0.05). In the combined racial group, the reliability coefficients, as measured by Cronbach's alpha, were 0.76, 0.78, 0.68, and 0.68 for the Impact, Loss, Stress, and Perceptions of Physician Efficacy scales, respectively. However, the results of within-racial-group analyses tell a different story. The Impact scale fitted the African American and white American subgroups, although there was some item variation by racial group. The Loss scale did not fit the white American subgroup, and the Stress and Perceptions of Physician Efficacy scales did not fit the African American subgroup. CONCLUSION: Perceptions of diabetes may vary by race, even when controlling for SES. Health professionals are in a unique position to develop diabetes management programs to help patients understand more about their diabetes, and thus to reduce some of the negative outcomes of diabetes by promoting adherence to recommended treatment regimens. In addition, overall measures of reliability may mask the instability of scales within specific study groups. PMID- 12772545 TI - Development of a culturally sensitive educational intervention program to reduce the high incidence of tuberculosis among foreign-born Vietnamese. AB - OBJECTIVE: In Los Angeles County and Orange County, 98% of Asians with tuberculosis (TB) were foreign born; newly arrived Vietnamese immigrants had the highest TB risk: i.e. > 100 times higher than that of the USA. The study objective was to find ways to reduce the high incidence of tuberculosis among the Vietnamese by: (1) identifying and understanding the cultural health beliefs and barriers among the Vietnamese population of Orange County, California; and (2) developing a partnership between UCLA/VA/RAND MEDTEP, key Vietnamese community based organizations, and community leaders in Orange County to develop and implement a comprehensive, culturally sensitive educational intervention program. DESIGN: Vietnamese residing in Orange County were recruited to obtain qualitative and quantitative data in 1998 and 1999, respectively. The study design included focus groups, in-depth interviews, and community surveys. Data were collected via qualitative reports, computer-assisted random telephone interviews, and self administered questionnaires. RESULTS: A conceptual schema was identified within the Vietnamese health belief system that recognized two different forms of tuberculosis: non-infectious psychological and infectious physical tuberculosis. It was possible to engage community organizations in developing programs to combat TB in the Vietnamese population. CONCLUSION: The Health beliefs of Vietnamese will be important considerations in developing a culturally sensitive educational intervention program to service this at-risk population. PMID- 12772546 TI - Recruitment of African Americans with type 2 diabetes to a randomized controlled trial using three sources. AB - OBJECTIVES: To assess the usefulness of three sources (faith-based organizations (FBOs), health system, and community) for recruitment of African Americans with type 2 diabetes to a randomized controlled trial (RCT). DESIGN: African Americans with type 2 diabetes were recruited to a diabetes self-management program at four FBO sites. An observational study of a multifaceted recruitment strategy to enroll subjects in the RCT that evaluated the effectiveness of a diabetes self management program and the effect of recruitment source on retention after enrollment. Self-administered demographic surveys and weekly class attendance records were collected. RESULTS: Of 184 interested individuals, 109 (59.2%) were enrolled. Of those enrolled, 60.6% recruited through the health system, 13.8% FBOs, and 19.2% the community. The highest yield was achieved through the health system. However, for both the intervention (I) and control (C) groups, respectively, participants recruited from FBOs (85.7% I; 62.5% C) were more likely to attend four or more sessions than those from the health system (75.0% I; 43.3% C) and community (55.6% I; 25.0% C). Despite similar class size, participants in the intervention group (74.5%; n = 41) were more likely to attend four or more of the seven classes than those in the delayed intervention (control group) (40.7%; n = 22). CONCLUSIONS: The findings suggest that African American adults with diabetes can be successfully recruited and retained in a racially targeted RCT conducted in FBOs. Key elements to consider are the use of a multifaceted approach for participant recruitment, particularly the benefit of health system physician involvement in recruitment since the highest yield was achieved through health system providers, and importance of site location for retention. PMID- 12772547 TI - Improving health outcomes in diverse populations: competency in cross-cultural research with indigenous Pacific islander populations. AB - OBJECTIVE: There is a large disparity in health status between the indigenous peoples of the US Associated Pacific compared to any population in the USA. The research process that has been supported by US academic institutions and federal agencies has been limited in its ability to address the disparate health issues and may be part of the problem. We define culturally competent research and review approaches to developing competency in cross-cultural research with indigenous Pacific Islander populations. DESIGN: This is a descriptive review of the investigators' experience in the Hawaii MEDTEP Center experience and of the experience of others conducting research with the indigenous people of the Pacific Islands. RESULTS: Culturally competent cross-cultural research with the indigenous peoples of the Pacific requires an understanding and application of indigenous peoples' paradigms of health, knowledge, science, and research. It is not sufficient to train more indigenous Pacific Islanders to do more Western style research. Unraveling the complex health situation and determining the changes that need to be made is dependent on the dominant culture engaging the indigenous Pacific populations in a way that bridges cultural paradigms. CONCLUSION: Positively affecting the disparity of health in the indigenous populations of the Pacific is, in part, dependent on employing an indigenous peoples-centered model of research. The model can have application to the study of indigenous peoples in other parts of the world. PMID- 12772548 TI - From publication to public action: Agency for Healthcare Research and Quality (AHRQ) perspectives on ethnicity and race-related outcomes research. PMID- 12772549 TI - [Mechanisms disordering wound healing on the lip after bilateral crossing of the inferior alveolar nerve and experimental validation of correction methods]. AB - The mechanisms of lip wound healing after bilateral crossing of the inferior alveolar nerve (IAN) were studied on Chinchilla rabbits in 3 experimental series, 6 animals per series. In group 1 bilateral crossing of IAN was carried out, in group 2 bilateral crossing of IAN was paralleled by removal of a mucous flap in the middle of the lower lip, and in group 3 the same wound as in group 2 was created, after which the wounds in this group were daily treated with a special ointment and a single injection of lidocaine (1% solution) under the wound. The nerve crossing led to development of ulcer on the lip with degenerative changes in the vascular walls, destruction of nerve fibers, and fragmentation of some axial cylinders. Crossing of IAN simultaneously with removal of the lower lip flap led to more severe degenerative changes in the tissue. Daily treatment of the lip with the ointment and lidocaine blocking normalized wound healing. A possible mechanism of the changes observed is discussed. PMID- 12772550 TI - [Evolution of tissue structures in the mandible after implantation of plate from polymethylmethacrylate and its compositions with hydroxyapatite]. AB - Tissue reactions to active modified composite material were studied in experiments on Chinchilla rabbits. In the first series of experiments plates from composite material were implanted: polymethylmethacrylate (PMMA) plates were used in group 1, PMMA composition with hydroxyapatite (HAP) in group 2, and PMMA composition with HAP modified by polyacrylic acid in group 3. The content of HAP in each group was 30% by weight. Implantation of plates from pure PMMA induced atrophic processes with formation of connective tissue capsule at the site of implantation. In the second series of experiments the manifestations of inflammatory reactions in the bone and adjacent tissues, caused by HAP release into tissue, were observed, which were paralleled by activation of bone formation, which sometimes acquired the pattern of a hyperproductive reaction. PMMA/HAP composite modified by chemical bond proved to be the most effective: its implantation caused no manifest pathological reactions and inflammatory infiltration; no intraosseous deposition of crystals were detected. Intensive osteogenesis round the implanted plate was observed. PMID- 12772551 TI - [Results of comparative studies of psychophysiological factors essential for self evaluation of pain during oral therapeutic interventions]. AB - The efficiency of infiltration, conduction, and intraligamental anesthesia with different anesthetics for oral therapeutic interventions was evaluated in 331 patients without concomitant somatic diseases. The levels of reactivity and personal anxiety, as well as the patient's mood have an essential impact on their own appraisal of the quality of analgesia, which the dentist should bear in mind when planning and carrying out anesthesia. Mean and high levels of anxiety are characterized by decreased subjective confidence in the quality of analgesia and can mislead the dentist, if he/she neglects these characteristics. PMID- 12772552 TI - [Efficacy of interval normobaric hypoxic stimulation in combined therapy of periodontal inflammations]. AB - Eighty patients with periodontal inflammations and 15 subjects matched for age and sex without periodontal inflammations were examined. Periodontal status was evaluated using clinical and paraclinical methods of examination. Blood content and oxygen saturation of periodontal tissues were evaluated with an Escort II MDE saturator (USA). The results indicate that interval normobaric hypoxic stimulation promoted a decrease of bacterial contamination of periodontal pouches and notably improved blood content and oxygen saturation of periodontal tissues. PMID- 12772553 TI - [Metabolic prerequisites for the development of dental diseases in pneumonia]. AB - The parameters of oral homeostasis were studied in patients with acute pneumonia before and after therapy. The redox potential, a shift of pH towards the acid values, decreased content of total protein, total and effective concentrations and reserve of albumin binding, and a drastic increase of toxicity index were detected. Analysis of oral fluid showed increased level of ionized calcium, nitrate anion, and decreased content of magnesium and ammonium cations. PMID- 12772554 TI - [Clinical laboratory characteristics, diagnosis, and treatment of acute odontogenic maxillary sinusitis]. AB - Clinical laboratory characteristics of acute odontogenic maxillary sinusitis are presented. All the patients (n = 132) were examined in accordance with a special diagnostic protocol. Two variants of acute inflammatory process were distinguished: serous and destructive. Conservative and surgical methods of treatment are suggested, depending on the disease pattern and complications. The treatment efficiency is compared to the results of traditional treatment. PMID- 12772555 TI - [Surgical methods used in the treatment of local gingival recession]. AB - Surgical methods used for repair of gingival recession are reviewed. Modern methods and means of plastic periodontal surgery solve many esthetic problems. The prognosis of the results of gingival plasty in cases of recession of different classes is now becoming more and more favorable. The method of purposeful tissue regeneration is a good supplement to surgical treatment of gingival recession. PMID- 12772556 TI - [Effect of ozone on antibiotic sensitivity of microorganisms]. AB - Ozone therapy of wounds of the face and neck notably increased the sensitivity of microorganisms to antibiotics in 79% cases. PMID- 12772557 TI - [Acute musculo-articular disorders in mandibular fractures of different location and methods of their correction]. AB - Ninety patients with acute musculo-articular disorders associated with mandibular fractures of different location were examined. Two groups of patients were distinguished, based on the presence of fragments dislocation, soft tissue injuries, and occlusion impairment: 1) 45 patients with mandibular fractures without essential dislocation of fragments and pronounced reaction of soft tissues and 2) 45 patients with notable impairment of soft tissues and dislocation of fragments. Clinical examinations and accessory diagnostic methods after removal of the splints demonstrated the relationship between the severity of musculo-articular disorders and type and location of mandibular fracture; purposeful methods of treatment have been developed. PMID- 12772558 TI - [A case with palatine torus of rare shape and size]. AB - Palatine torus is a bone formation on the palate at the site of the palatine suture, a result of connection of the palatine mandibular processes grown together. The shape and size of the palatine torus vary. Large size of the palatine torus makes difficult the designing of removable dentures for orthodontic treatment. PMID- 12772559 TI - [Guying denture systems. Part 1. Tooth splinting]. AB - One of the elements of gyuing dentures suggested by the author is described in detail. It serves for redistribution of masticatory loading between natural teeth and can be used alone (for tooth splinting) and in combination with other elements of the proposed denture system (for orthodontic treatment of partial defects of dentition). The schemes of splinting and their biomechanics are described in detail. High reliability of the proposed design is emphasized. PMID- 12772560 TI - [Effects of removable plate dentures made of acrylic plastic on the structure and function of oral mucosal cell membranes]. AB - The results of orthodontic treatment of 120 patients with complete and partial adentia are presented. The patients were fitted with removable plate dentures on the base of acrylic polymers trevalon, fluorax, ethacryl, and redont. The results of biochemical and morphological studies recommend the basic material trevalon for practical orthodontics. Measurements of medium molecular weight oligopeptides, histamine, malonic dialdehyde, and activities of antioxidant enzymes are recommended for predicting the time and quality of patients' adaptation to removable plate dentures from acrylic polymers. PMID- 12772562 TI - [Methods for evaluating tooth mobility]. PMID- 12772561 TI - [Time of tooth eruption in children born preterm in the arid climatic zone]. AB - Dental examinations were carried out in 2768 preschool children in the city of Tashkent. 407 (14.7%) of these children were born preterm. The time of tooth eruption was evaluated from analysis of 295 case histories and 112 infants were observed from the age of 3 months till 2 years. The terms of decidual teeth eruption in preterm children are different in comparison with children born after full-term gestation and depend on the baby's maturity by birth. Diseases of the first months of life have a negative impact on the time of tooth eruption. PMID- 12772563 TI - [Mechanisms of preventive and therapeutic action of permanent magnetic field and magnetic laser radiation]. PMID- 12772564 TI - [Motivation aspects of patients' application for care under conditions of a territorial system of dental care organization]. PMID- 12772565 TI - [The present and the past of dentistry service in the Republic of Tajikistan]. PMID- 12772567 TI - [Pathology of gastric MALT lymphoma]. PMID- 12772566 TI - [Molecular mechanism for hematogenous metastasis of gastrointestinal cancer and its clinical application for therapy]. PMID- 12772568 TI - [Diagnosis and treatment of gastric MALT lymphoma]. PMID- 12772569 TI - [An economic evaluation on the clinical pathway for gastrectomy]. AB - OBJECTIVE: To analyze the costs of gastrectomy patients treated with the clinical pathway. PATIENTS AND METHODS: Seventy-six patients (path group 44, control 32) had undergone gastrectomy in our hospital in 2001. The clinical pathway included the same care map. Treatment costs were estimated from medical cost receipt data. The economical analysis was performed from the point of the direct cost payer's view. RESULTS: The length of hospital stay in the path group was 27.1 +/- 5.9 days and decreased 8.3 days in comparison with the control(p < 0.001). The cost of the path group was 145.290 +/- 23.773 points and 19.278 points less than the control(p < 0.005). In the path group the operation case per bed was increased 30% and the cost per bed was also increased 15% more than the control. CONCLUSIONS: The implementation of the clinical pathway decreased the length and the cost of hospital stay. The clinical pathway is effective to use the hospital resources, such as bed. PMID- 12772570 TI - [A case of esophageal pyogenic granuloma which showed change of form in a short term]. PMID- 12772571 TI - [A case of refractory ulcerative colitis treated successfully with a combination of low-dose heparin and 5-aminosalicylic acid]. PMID- 12772572 TI - [A case of gastrointestinal stromal tumor of the small intestine, preoperatively diagnosed by the enteroscopy]. PMID- 12772573 TI - [A case of catheter tip dislocation to the duodenal bulb for hepatic arterial infusion chemotherapy]. PMID- 12772574 TI - [A case of early syphilitic hepatitis with jaundice]. PMID- 12772576 TI - [A case of biliary tract cancer shown rare progress and development]. PMID- 12772575 TI - [A resectable case of hepatocellular carcinoma complicated with hepatic alveolar echinococcosis]. PMID- 12772577 TI - [A case of splenic abscess associated with pancreatic tail cancer]. PMID- 12772578 TI - [A case of abdominal wall actinomycosis]. PMID- 12772579 TI - [A case of peritoneal malignant mesothelioma]. PMID- 12772580 TI - Tissue reactions after intraosseous implantation of three retrofilling materials. AB - Bone tissue reactions to EBA, IRM, and cyanoacrylate cement (Base Liner) were studied in the rat mandible using an intraosseous implant method. Osseous cavities (1.4 mm in diameter) were surgically created in the mandibles, and materials were implanted in 60 male Wistar rats. Each specimen was evaluated histologically after 4 and 8 weeks. The development of fibrous connective tissue in direct apposition to the material was observed in the EBA and IRM groups at 4 weeks. A slight degree of macrophage infiltration was seen in the EBA group. After the 8-week observation period, IRM and EBA were frequently separated from the bone cavity by a fibrous connective tissue layer (p < 0.01). The Base Liner appeared to be in direct apposition to the osseous tissue in several areas (p < 0.01). These findings indicate that Base Liner reacts favorably with osseous tissue, compared with the EBA and IRM materials tested and seems to be a biocompatible material. PMID- 12772581 TI - Effect of chitosan rinsing on reduction of dental plaque formation. AB - The purpose of the present study was to investigate whether the use of a chitosan mouthrinse could be efficacious in reducing plaque and saliva mutans streptococci level. A randomized crossover clinical trial was performed to evaluate the effect of a rinse with 0.5% chitosan for 14 days on plaque formation and mutans streptococci counts in saliva. Twenty-four subjects were randomly assigned either the chitosan rinse or a placebo rinse in addition to their usual oral hygiene procedures. Following the baseline examination, each subject was given a prophylaxis. They were instructed to rinse with 20 ml of the mouthrinse twice daily for 30 seconds. Plaque scores were measured after a 14-day rinsing period, and mutans streptococci counts in saliva were also determined at the start and the end of the each rinsing period. The procedures were repeated with the alternate rinse after a 14-day washout period. Rinsing with 0.5% chitosan was significantly more effective in plaque reduction using the Quigley & Hein Index (chitosan: 1.44, placebo: 1.62, p < 0.001) and Plaque Severity Index (chitosan: 0.138, placebo: 0.186, p = 0.003). The mutans streptococci count in saliva was less after the chitosan rinsing (chi 2 cal = 13.51, p = 0.035) than placebo rinsing. In conclusion, the chitosan rinsing was effective in reducing plaque formation and counts of salivary mutans streptococci after a 14-day rinsing period. These results would appear to warrant further investigation into the potential value of chitosan as an effective anti-plaque agent for use in oral hygiene products. PMID- 12772582 TI - A juvenile case of cerebellar arteriovenous malformation (AVM) with gradual onset of headache and ataxia. AB - An 11-year-old male was admitted because of frequent vomiting and truncal ataxia which had lasted for over one week. He had clear consciousness but slowly progressive mild headache and ataxic gait. Cranial CT revealed a 4 cm hematoma in the right cerebellar hemisphere. Angiography showed a 2 x 2 cm nidus of a pial arteriovenous malformation (AVM) in the right hemisphere fed from the right posterior inferior cerebellar artery and draining into the inferior hemispheric vein. We performed a surgical resection of the AVM after decompression therapy to counteract the brain edema. He recovered completely without any neurological deficits. This case suggests that cerebellar hemorrhage caused by AVM should be considered as a possible diagnosis when mild symptoms of headache and ataxia proceed gradually. PMID- 12772583 TI - PACAP-induced depolarizations in hamster submandibular ganglion neurons. AB - In this study, we have investigated the effects of pituitary adenylate cyclase activating polypeptide (PACAP) on in vitro hamster submandibular ganglion neurons using the conventional intracellular recording technique. PACAP (10 microM) induced slow depolarizations in approximately 70% of tested cells. PACAP-induced depolarizations were approximately 10 mV in the peak amplitude, and their durations were approximately 10 min. The slow depolarizations were accompanied by a decrease in membrane conductance (gm) at the initial phase and an increase in gm at the peak phase. Membrane input resistance increased by 14.8 +/- 2.2% (mean +/- S.E., max.) of the resting value at the initial phase and decreased by 30.8 +/- 4.3% (max.) at the peak phase. Anodal break spikes were elicited at the initial phase during PACAP-induced depolarization. In one neuron, anodal break spikes were elicited at the peak. Spikes which followed the anodal break spike were also elicited at 4 Hz in the initial phase during the slow depolarizations. The decrease in gm was probably produced by an inhibition of calcium conductance and an inhibition of slow Ca(2+)-activated K+ channels, while the increase in gm might have been produced by an activation of nonselective cation channels. The slow depolarizations by PACAP might be mediated by a membrane-delimited signal transduction cascade involving G protein in the submandibular ganglion neurons. PMID- 12772584 TI - [Studies on the syntheses of the hypermodified nucleosides of phenylalanine transfer ribonucleic acids]. AB - This review highlights the first total synthesis of (alpha S, beta S)-beta hydroxywybutosine, the identity of which was established with fluorescent nucleoside isolated from rat liver tRNA(Phe). Three general prerequisites for the synthesis of the target are emphasized: construction of new 9-beta-D ribofuranosyl-3-methylpurines; the first syntheses of optically active gamma-aryl alpha,beta-unsaturated amino acids; and a new method for hydrogenolysis of glycols through their cyclic carbonates to give the gamma-aryl-beta-hydroxy amino acid derivatives. The mechanism for the formation of cyclic carbonates from reactions of glycols with (COCl)2 and highly selective preparation of the cyclic oxalates are also described. PMID- 12772585 TI - [Development of carbon radical addition to imine derivatives]. AB - This review summarizes the new carbon-carbon bond construction methods based on the radical reaction of imine derivatives. The intermolecular carbon radical addition to oxime ethers proceeded smoothly in the presence of BF3.OEt2. A high degree of stereocontrol in the reaction of oxime ethers was achieved to give amino acid derivatives with excellent diastereoselectivities. The radical reaction of imine derivatives in water has also been investigated. The radical cyclization of oxime ethers proceeded effectively to provide the functionalized heterocycles via a carbon-carbon bond-forming process. These reactions were extended to the solid-phase radical reactions using triethylborane or diethylzinc as a radical initiator. PMID- 12772586 TI - Regulation by prostaglandin E2 and histamine of angiogenesis in inflammatory granulation tissue. AB - In an air pouch-type carrageenin-induced inflammation model in rats, the selective cyclooxygenase (COX)-2 inhibitor NS-398 dose dependently inhibited the granulation tissue formation, angiogenesis and the level of vascular endothelial growth factor (VEGF) in the granulation tissue. In culture of the minced granulation tissue, PGE2 induced VEGF production in a concentration-dependent manner. Histamine also induced VEGF production in the granulation tissue in vitro. The H2 receptor antagonist cimetidine, the cAMP antagonist Rp-cAMP and the protein kinase A inhibitor H-89 suppressed the histamine-induced VEGF production in the granulation tissue. However, the H1 receptor antagonist pyrilamine maleate, the H3 receptor antagonist thioperamide, the protein kinase C inhibitors Ro 31-8425 and calphostin C or the tyrosine kinase inhibitor genistein showed no effect. Subcutaneous implantation of a cotton thread in the dorsum of histidine decarboxylase-deficient (HDC-/-) mice, but not in mast cell-deficient (WBB6F1 W/Wv) mice, induced less angiogenesis with lower levels of VEGF in the granulation tissue than in their corresponding wild-type (HDC+/+ and WBB6F1( )+/+) mice. In HDC-/- mice, the topical injection of histamine or the H2 receptor agonist dimaprit rescued the defective angiogenesis and granulation tissue formation. In addition, cimetidine but not pyrilamine maleate and thioperamide inhibited the histamine-induced angiogenesis in the granulation tissue in HDC-/- mice. These findings suggest that PGE2 and histamine augment angiogenesis in the inflammatory granulation tissue by inducing VEGF production, and histamine induces VEGF production possibly through the H2 receptor--cAMP--protein kinase A pathway. PMID- 12772587 TI - [Spectroscopic analyses of the interaction between hammerhead ribozymes and metal ions]. AB - Biologically active RNA molecules utilize metal ions to fold into specific conformations or to form a catalytic center. Hammerhead ribozymes also contain a metal ion-binding motif in their conserved core region. This motif and bound metal ions have recently been focused on from the viewpoint of whether the metal ions are a catalytic metal or a structural metal. We studied the interaction between metal ions and an RNA oligomer, r(GGACGAGUCC), which mimics the metal ion binding motif of hammerhead ribozymes, using NMR spectroscopy in solution. Using 15N-labeled RNA oligomers, the chemical shift of N7 of the G7 residue [N7(G7)] in the metal ion-binding motif was specifically perturbed upon the addition of Cd(II). It was also found that the 31P resonance of the phosphate of the A9 residue showed the largest perturbation of all the 31P resonances during titration. These data indicate that divalent cations can bind to the metal ion binding motif in solution, and that the binding sites for cations are N7(G7) and the phosphate of A9. More importantly, the metal ion-binding motif is an independent functional module that can capture divalent cations without the assistance of other conserved residues in hammerhead ribozymes. PMID- 12772588 TI - [Studies on the new antiarthritic drug candidate S-2474]. AB - Various 1,2-isothiazolidine-1,1-dioxide (gamma-sultam) derivatives containing an antioxidant moiety, 2,6-di-tert-butyl-phenol substituent, were prepared. Some compounds that have a lower alkyl group at the 2-position of the gamma-sultam skeleton showed potent inhibitory effects on both cyclooxygenase (COX)-2 and 5 lipoxygenase (5-LO), as well as production of interleukin-1 (IL-1) in in vitro assays. They also proved to be effective in several animal arthritic models without any ulcerogenic activities. Among these compounds, (E)-(5)-(3,5-di-tert butyl-4-hydroxybenzylidence)-2-ethyl-1,2- isothiazolidine-1,1-dioxide (S-2474) was selected as an antiarthritic drug candidate. The structure-activity relationships examined and some pharmacological evaluations are described. Furthermore, we have developed an efficient and E-selective synthesis of S-2474, in which alpha-methoxy-p-quinone methide is used as a key intermediate. alpha Methoxy-p-quinone methide was revealed to be equivalent to a p-hydroxy-protected benzaldehyde. It reacts smoothly with alpha-sulfonyl carbanion to give 1,6 addition intermediates, which can be further processed to provide S-2474 directly in the presence of a base. This procedure gives S-2474 as an almost single isomer on the benzylidene double bond in excellent yield and thus is a very practical method adaptable to large-scale synthesis. The detailed mechanistic aspects are studied and discussed. PMID- 12772589 TI - [Establishment and evaluation of a system for preventing mis-administration of powder using bar codes printed on drug envelopes]. AB - Even if drugs are accurately dispensed, it is difficult to ensure their safety in the absence of an error-free medication system when or after drugs are given to patients. In particular, most powders are white and are impossible to distinguish based on appearance after they are placed in powder packages. In this study, we newly developed a system in which prescription order information is bar-coded on drug envelopes, and read when powder is folded, facilitating efficient and accurate printing of the patient's name and drug name on powder packages. Seventy one (86%) of 83 nurses surveyed indicated that the system was useful for resolving high-risk problems they had experienced: "when powder was taken from the envelope, the contents were unknown", and "the powder was administered to another patient." PMID- 12772590 TI - [Structure activity relation study of matrine-type alkaloids. Part III]. AB - 6-Acyldecahydro [1,6]naphthyridines were synthesized as derivatives of matrine type and allomatrine-type alkaloids, and the structure-activity relations were examined by the acetic acid-induced abdominal contraction test. All synthesized derivatives produced the antinociception in mice. The antinociceptive potencies of 15a-c and 16a-c were lower than those of 17a-c, 18a-c, 19a-c and 20a-c. Furthermore, those of the matrine-type derivatives 17b and 17c are greater than other derivatives. These findings suggest that less hindered tertiary amine and highly lipophilic acyl group are better functional groups for the greater antinociceptive potencies. Furthermore, these findings suggest that A or B ring of 1 and 2 are not essential for the antinociceptive effect. PMID- 12772591 TI - [Precision of internal standard method in HPLC analysis]. AB - The internal standard methods are known to compensate for the errors from sample preparation and injection into an analytical instrument. However, recent HPLC apparatuses have injectors of excellent repeatability and it is dubious whether the cancellation of injection error can lead to substantial improvement in the precision of analysis. This paper answers the above question experimentally and theoretically. The HPLC analysis of butylscopolamine bromide is taken as an example. The relative standard deviations (RSD) of measurements in the internal standard method and absolute calibration curve method are compared and the advantages of these methods are discussed. The measurement RSD is shown to be well estimated by the (function of mutual information) (FUMI) theory without repeating measurements. This report also demonstrates simple equations for calculating the measurement RSD at an arbitrary concentration of analyte and for selecting the better method between the internal standard method and absolute calibration curve method under specific experimental conditions. PMID- 12772592 TI - [The association between dispensing error factors and behavioral characteristics of pharmacists]. AB - We evaluated error prevention education by clarifying the association between dispensing error factors and behavioral characteristics of pharmacists. The subjects were 98 pharmacists (27 men and 71 women) with a mean age of 29.7 years who gave informed consent for participation in our survey. Between November 2001 and January 2002, a questionnaire survey on dispensing errors was performed using the "Tokyo University Egogram, New Version" for the assessment of behavioral characteristics and the "Safety Behavior Questionnaire" for the assessment of error factors. An association was observed between the incidence of dispensing errors and behavioral characteristics. There was also an association between error contents and behavioral characteristics as well as error factors. With more experience, errors associated with becoming accustomed increased, suggesting that error prevention education is necessary not only for newly qualified pharmacists but also for managers. PMID- 12772593 TI - [A case in which tofisopam was effective for treatment of paroxysmal supraventricular tachycardia]. AB - Paroxysmal supraventricular tachycardia (PSVT) is a reentrant tachycardia, and the autonomic nervous system influences the paroxysms of PSVT. Tofisopam (Grandaxin) is a derivative of benzodiazepines. It is reported that heart rate variability (HRV) reveals the function of the autonomic nervous system and is used as an index of the effects of the autonomic nervous system on the heart rhythm. In previous studies, it was reported that tofisopam improved HRV. In this case report, the frequency of PSVT was significantly decreased and subjective symptoms of arrhythmia were eliminated after the administration of tofisopam. R-R interval variability and high-frequency power (HF; 0.15-0.40 Hz) were increased, and [low-frequency power (LF; 0.04-0.15 Hz)]/HF was decreased after administration of tofisopam. These results suggest that tofisopam overcame the existing imbalance of the sympathetic and vagus nervous system. In this case report, it is suggested that tofisopam is effective for the treatment of PSVT. PMID- 12772594 TI - Tissue distribution of mRNA expression of human cytochrome P450 isoforms assessed by high-sensitivity real-time reverse transcription PCR. AB - Pairs of forward and reverse primers and TaqMan probes specific to each human cytochrome P450 isoform were prepared. Analysis of the mRNA level of each CYP isoform in total RNA from pooled specimens of various human organs was performed by real-time reverse transcription PCR using an ABI PRISM 7700 sequence detector system. The expression of CYP3A4 mRNA was similar to that of CYP3A7 mRNA in the fetal liver, and CYP3A4 mRNA levels in the fetal liver were about 0.1 times lower than in the adult liver. CYP2E1 showed the highest level of mRNA expression in the liver. The mRNA expression of 30 CYP isoforms (CYP1A1, 1A2, 1B1, 2A6, 2A7, 2B6, 2C8, 2C9, 2C18, 2C19, 2D6, 2E1, 2J2, 3A4, 3A5, 3A7, 4A11, 4F2, 4F3, 5A1, 7B1, 8A1, 8B1, 17, 26A1, 27, 27B1, 39A1, 46, and 51) in the liver was successfully detected by this method. CYP2F1, 4B1, 4F8, 11s (11A, 11B1, and 11B2), 19, and 24 mRNA levels were the highest in the lung, lung, prostate, adrenal gland, placenta, and kidney, respectively; however, the mRNA expression of these eight CYP isoforms in the liver was not detected by this method. The mRNA levels of the CYP isoforms determined in various human tissues were in good agreement with previously reported data. The method described here has the advantages of high specificity and excellent quantification over a wide range of mRNA concentrations, making it suitable for the evaluation of a large number of samples in the assessment of the expression profile of drug-metabolizing enzymes. PMID- 12772596 TI - [A case of M. avium lung disease complicated with adenocarcinoma]. AB - A 56-year-old man was admitted to our hospital for examination of an abnormal shadow found in a chest radiograph. Chest CT demonstrated a thick-walled cavity surrounded by small centrilobular nodules in the upper lobe of the left lung. Chemotherapy with rifampin, isoniazid, ethambutol and pyrazinamide was started, because acid-fast bacilli were observed in the bronchial brushing smear. Since PCR examination of the bronchial lavage was positive for M. avium, the rifampin and ethambutol were retained, while the other drugs were replaced with streptomycin and clarithromycin. However, in this case the radiographic findings did not point to non-tuberculous mycobacterial (NTM) infection, since the thickness of the cavity was irregular in the caudal portion. In addition, the patient did not accept long-term NTM therapy. Therefore, we chose surgical therapy. In a portion of the surgical specimen, adenocarcinoma was detected. The possibility of lung cancer should be remembered in cases with a thick-walled lung cavity. PMID- 12772595 TI - [Telemedicine support system in home care of patients with chronic respiratory failure: preliminary results]. AB - We report the preliminary results of an advanced telemedicine system for the delivery of home care services. We have been trying out a system for a ternary telemedicine network for remote consultation in patients with chronic respiratory failure. The elements of this system are: the patients at home, the home doctor (and/or community hospital), and an institution with respiratory staff (Shinshu University), which were linked by ISDN lines. The system allows real-time visualization, at home, of the patients and their physiological data, which are transmitted simultaneously to both the community hospital and Shinshu University. Respiratory specialists can advise the home doctor on the treatment and follow-up evaluations of distant patients. We consider that the telemedicine network system has the potential to improve clinical outcomes and to provide home care services to patients with chronic health conditions. PMID- 12772597 TI - [A case of pulmonary actinomycosis with recurrent hemoptysis]. AB - A 68-year-old man was admitted to our hospital because of hemoptysis in September 1999. Chest CT scans showed a nodular shadow with infiltration in the right S 7. Bronchial arteriography showed vascularization in the right S 7, and bronchial artery embolization was performed. However, in April and October 2000 hemoptysis recurred, and bronchial arteriography showed recurrence of vascularization in the same area, so embolization was performed again. Then, the patient was admitted in March 2001 because of recurrent hemoptysis. CT scans showed growth of the nodular shadow. Right lower lobectomy was performed, and the microscopic findings in the tissue from the resected lobe showed branching filamentous bacteria, and pulmonary actinomycosis was diagnosed. We concluded that pulmonary actinomycosis should be considered in the differential diagnosis of nodular shadows with recurrent hemoptysis. PMID- 12772598 TI - [Paragonimiasis westermani caused by ingestion of Chinese freshwater crabs]. AB - We encountered three cases of paragonimiasis westermani caused by ingestion of Chinese freshwater crabs. All patients are Chinese living in Japan who had a history of eating raw Shanghai crabs at the Chinese restaurant owned by one of them. A few months later they complained of cough, chest pain and dyspnea. On examination, their peripheral blood showed a remarkable increase in eosinophils. Chest radiographs revealed the presence of pleural effusion in all cases, and of pneumothorax in one. Pleural fluid obtained from two cases showed an increased number of eosinophils, an elevated LDH, and a decreased glucose level. Paragonimus westermani was determined in a positive serum antibody test. The patients were treated with praziquantel for 3 days at a daily dosage of 75 mg/kg and their symptoms were relieved, but one case needed additional treatment with praziquantel for 3 days at a daily dosage of 50 mg/kg because of sustained pleural effusion and a persistently high antibody titer. Most cases of paragonimiasis found in Japan have been sporadic and individual, but three cases occurring close to each other in both distance and time have recently been reported. Such multiple infections are likely to increase because of the gourmet food boom and the internationalization of eating habits. Paragonimiasis must be kept in mind in the differential diagnosis of cases of pleural effusion with eosinophilia. We would therefore like to emphasize the importance of the detection of parasite-specific antibodies and of careful history-taking from patients. PMID- 12772599 TI - [A case of intrapulmonary teratoma presenting with expectoration of white hair and cheese-like material]. AB - A 41-year-old man presented with expectoration of white hair and cheese-like material. The chest radiograph showed a round shadow adjacent to the right hilum. Bronchofiberscopy revealed strands of white hair and an obstruction consisting of sebaceous material in the right B3b bronchus. Right upper lobectomy was performed. Histological examination confirmed the diagnosis of intrapulmonary mature teratoma. This characteristic clinical presentation and the bronchofiberscopic findings may have been specific for the diagnosis of teratoma in this case. PMID- 12772600 TI - [A case of idiopathic pulmonary upper lobe fibrosis complicated by invasive pulmonary aspergillosis]. AB - A 72-year-old man with idiopathic pulmonary upper lobe fibrosis who had been followed for a year developed a high fever and yellow sputum in July 2001. Chest radiography and chest computed tomography (CT) showed a rapidly enlarging cavity with an internal mass and infiltration in the left upper lung field. Pulmonary aspergillosis was diagnosed by examination of bronchoalveolar lavage fluid (BALF). Administration of itraconazole improved his condition. The concentrations of surfactant proteins A (SP-A) and D (SP-D) in serum and in BALF were decreased during the clinical course. It is known that SP-A and SP-D are critical factors for host defense against aspergillus. The lowering of SP-A and SP-D in the serum and BALF seemed to reflect destructive changes of lung structure and impaired innate lung immunity that could to lead invasive pulmonary aspergillosis. PMID- 12772601 TI - [Pulmonary actinomycosis diagnosed through transbronchial lung biopsy (TBLB)]. AB - Pulmonary actinomycosis is a rather rare chronic pulmonary infection. Its diagnosis may be difficult without biopsy, because its culture from sputum or bronchial secretion is rarely successful. We report a case of pulmonary actinomycosis diagnosed by means of TBLB. A 57-year-old man was admitted to our hospital because of hemosputum. The chest radiograph at the first visit revealed a nodular lesion in the right middle lobe. The biopsy examination showed sulfur granules of which PAS-positive mycelium was arranged in a radiating pattern. On the basis of these findings, we diagnosed pulmonary actinomycosis. So far, there have been only 4 reported cases of pulmonary actinomycosis diagnosed by TBLB in Japan. TBLB is worth trying in the diagnosis of pulmonary actinomycosis, because it can avoid surgical procedures, if successful. PMID- 12772602 TI - [A case of sarcoidosis with rheumatic features (Lofgren's syndrome)]. AB - Rheumatoid arthritis was diagnosed in a 30-year-old woman with erythema nodosum and arthritic symptoms since 1994, and she was treated with anti-rheumatic agents. Mediastinal and bilateral hilar lymphadenopathy and abnormal pulmonary shadows were detected in 1996, and she was admitted to our hospital in 1997. We also recognized the elevation of ACE and lysozyme, and found granulomas in a transbronchial lung biopsy and an arthrosis synovia biopsy. From these findings, sarcoidosis was diagnosed. Sarcoidosis demonstrating erythema nodosum, arthritis, and bilateral hilar lymphadenopathy is called Lofgren's syndrome. In Caucasians, Lofgren's syndrome is frequently encountered, but it is rare in Japanese. Our case had coexisting arthrosis symptoms, and satisfied the diagnosis criteria of rheumatic arthritis. Therefore, the differential diagnosis was important. We emphasize that it is necessary to consider Lofgren's syndrome when diagnosing patients with rheumatic features, even in Japan. PMID- 12772603 TI - [Two cases of Legionella pneumophila pneumonia improved by parenteral ciprofloxacin administration]. AB - We report here two cases of Legionella pneumophila pneumonia that were markedly improved by parenteral ciprofloxacin administration. A 69-year-old man who had previously visited a hot spring was admitted to our hospital with severe pneumonia and a 48-year-old man with dilated cardiomyopathy as an underlying disease was also hospitalized because of heart failure and pneumonia. In both cases a urinary antigen test for L. pneumophila was negative at the incipient stage, and the initial treatment with a beta-lactam agent was ineffective. However, the high titer of L. pneumophila serogroup 6 antigen in the serum at the convalescent stage revealed that these two pneumonia cases were caused by L. pneumophila, and the following intravenous administration of ciprofloxacin was highly effective. We concluded that intravenous treatment with ciprofloxacin could be effective against L. pneumophila pneumonia, which is sometimes hard to diagnose in the acute phase. PMID- 12772604 TI - [A case of paraneoplastic syndrome improved following chemoradiotherapy for lung cancer]. AB - Paraneoplastic neurosyndrome (PNS) is a group of neurological disorders caused by or associated with neoplasms that are not direct effects of the primary tumor or of a metastasis to the involved organs. Chemotherapy, radiotherapy, immunosuppressive therapy, and plasmapheresis have been performed to treat PNS, but improvement of the neurological disorder is rather rare. A 64-year-old man was referred to our hospital with dysesthesia of the extremities and ataxic gait. Small cell lung cancer was diagnosed in another hospital and chemotherapy (CDDP 80 mg/m2 + VP-16 100 mg/m2) was performed. A partial response was obtained with this treatment, but the neurological dysfunction was exacerbated. Three months later, the patient was admitted to our hospital. On treatment with CDDP 80 mg/m2 (day 1) and CPT-11 80 mg/m2 (days 1, 8 and 15) and subsequent radiation therapy (60 Gy), his neurological disorder improved. We consider that neurological symptoms are important signs of malignancy in PNS and that a full course of treatment could improve the neurological disorders. PMID- 12772605 TI - [A case of juvenile pulmonary infarction associated with diet therapy]. AB - A 31-year-old man experienced chest pain, fever, bloody sputum and cough after diet therapy. Chest radiography and chest CT showed infiltration in the right lower lung field and right pleural effusion. Pulmonary embolism and infarction was diagnosed using 99mTc-MAA perfusion scans and chest enhanced CT. The patient did not have a thrombotic disposition and deep vein thrombosis in the lower extremities. This case did not have an acute onset or dyspnea, and was not typical of pulmonary embolism. The diet therapy may have caused dehydration and acted as a predisposing cause of pulmonary embolism. PMID- 12772607 TI - [Two cases of allergic bronchopulmonary mycosis caused by Schizophyllum commune in young asthmatic patients]. AB - We report on two patients, a 27-year-old and a 33-year-old woman, with allergic bronchopulmonary mycosis (ABPM) caused by the basidiomycetous fungus Schizophyllum commune (S. commune). Each patient had bronchial asthma. Both were admitted to our institution for further examination of cough, sputum, and abnormal chest shadows. ABPM was strongly suspected, because they showed eosinophilia in both peripheral blood and sputum, and increased serum IgE levels. A mold was isolated from their sputum, but identification was not possible. Systemic corticosteroid therapy relieved their symptoms and chest abnormal shadows. Later, S. commune, a basidiomycetous fungus, was detected from further examination of their sputum cultures, and serum anti-S. commune IgG was elevated. Finally, both cases were diagnosed as ABPM caused by S. commune. It is reported that this syndrome typically develops in women in middle age, but our patients were young women. It is important to take into account the possibility of ABPM caused by S. commune even in young patients when Aspergillus species are not isolated. PMID- 12772606 TI - [A case of HTLV-1-associated bronchiolitis with Sjogren's syndrome]. AB - A 58-year-old woman was admitted because of bilateral diffuse nodular shadows on chest radiograph. Video-assisted thoracoscopic lung biopsy specimens showed marked lymphocytic infiltration into the bronchiolar wall. HTLV-1 antibody was examined because the patient's parents were born in Okinawa Prefecture. The patient turned out to be an adult T-cell leukemia-lymphoma (ATLL) carrier with positive HTLV-1 antibody. Therefore, HTLV-1 associated bronchiolitis was diagnosed. Although no dry symptoms appeared, it became clear by the saxon test that she was also complicated with Sjogren syndrome. Generally, it is reported that there is coherence with HTLV-1 infection and Sjogren syndrome. So the HTLV-1 infection might have accounted for the pathogenesis of these three disorders (HTLV-1 infection, bronchiolitis, Sjogren syndrome). PMID- 12772608 TI - [A case of unification anomalies of the superior vena cava and absence of the left brachiocephalic vein with dilated azygos and hemiazygos veins]. AB - We report a case of dilated azygos vein simulating a tumor in the posterior mediastinum. The patient was a 47-year-old male who was found to have anomalies of unification of the superior vena cava and absence of the left brachiocephalic vein. He was admitted to our hospital because chest radiographs and computed tomography (CT) scans showed a well-circumscribed oval mass 2.5 cm in diameter, behind the bifurcation of the trachea. This lesion, which was markedly enhanced in chest CT, magnetic resonance imaging (MRI) and MR angiography (MRA) was diagnosed as a dilated azygos vein. An enlarged hemiazygos vein and collaterals were also recognized. Upon digital subtraction venography of both upper extremities, the right atrium and the left brachiocephalic vein could not been distinguished from the superior vena cava and the infraclavicular vein, respectively. Collaterals entering the azygos and hemiazygos veins were, however, recognized. It was considered that MRA and disital subtraction venography were very useful for confirmation of the diagnosis in this case. PMID- 12772609 TI - [Lifestyle differences between conjugal bereaved women and non-bereaved women in later life]. AB - PURPOSES: In Japan, there has been limited research regarding lifestyle of elderly bereaved. The present study was conducted to clarify lifestyle characteristics in conjugal bereaved women compared to non-bereaved women. METHODS: Subjects consisted of 872 bereaved and 643 non-bereaved elderly women aged 65 or over in T prefecture. Bereaved subjects experienced the death of a spouse in 1994 and their family structure remained unchanged after the bereavement. Non-bereaved subjects were randomly selected from community registration lists in 1995 and matched for age and area of residence. Lifestyle characteristics were obtained by home visit and interview. For the analysis, we divided the subjects into 4 groups by age and family structure. Multiple logistic regression analysis was performed to identify lifestyle characteristics of bereaved elderly women, after adjustment for physical ability and disease, as confounding factors. RESULTS: Bereaved elderly women were significantly associated with no roles at home, no hobbies, lack of contact with friends, physical inactivity, sleeplessness and irregular meal patterns compared to the non-bereaved. Analysis of the 4 groups revealed bereaved women living with family members have more unfavorable lifestyle characteristics than bereaved women living alone. Bereaved women living with family members had no roll at home and sleeplessness. CONCLUSION: Bereaved women have unfavorable lifestyle characteristics. Those living with family members had more unfavorable lifestyle than bereaved women living alone. From the preventive point of view, interventions for bereaved women are needed. This is particularly the case for bereaved women living with family members. Not only health services but also social welfare services are needed to reconstruct favorable lifestyles and maintain health and well-being. PMID- 12772610 TI - [Health-related lifestyle and mental health among inhabitants of a city in Japan]. AB - AIM: Few studies have linked mental health to lifestyle factors in the Japanese general population. The present investigation was conducted to examine whether mental health is associated with a health-related lifestyle and how the strength of the association, if any, differs across sex and age groups, using health survey data for inhabitants of Japan. METHODS: The data used were obtained from a questionnaire survey on lifestyle and health. In 1998, the questionnaires were mailed to 2,288 subjects selected from among the inhabitants of a city in Japan, of whom 1,642 subjects (71.8%) responded. The present study included 1,343 subjects who completed all the items that were used in the analysis. The General Health Questionnaire (GHQ) -12 was used as an indicator of mental health, with 4 points or higher (high score) indicating poor mental health. Information on eight different lifestyle parameters was also obtained. Logistic regression was used to calculate the odds ratio (OR) of having poor mental health for each lifestyle. RESULTS: Subjects in poor mental health had a lower mean score for a healthy lifestyle than those in good mental health. The difference in the mean score increased with age among men, while it decreased with age among women. The OR for having poor mental health among non-exercisers was higher in the older age group among men, while it was higher in the younger age group among women. A strong association between sleep hours and mental health was observed in the older age group among men, while it was seen in the younger age groups among women. Dietary factors in general showed a clearer association with mental health among women than among men, except for the consumption of salty foods, the association of which with mental health was more evident among men. CONCLUSIONS: Mental health was found to be significantly associated with health-related lifestyles in the Japanese population. The strength of the association for each lifestyle varied considerably across sex and age groups. PMID- 12772611 TI - [Resource utilization of gastric cancer patients aggregated item-by-item and comparison of the medical costs with the reimbursement level and length of hospital stay]. AB - OBJECTIVE: Using the new hospital cost accounting method per case based on cost per service, we compared the medical costs with the reimbursement level and length of hospital stay for gastric cancer patients. METHOD: The subjects were 158 gastric cancer patients who were admitted for surgery in a public hospital in Tokyo between 1995 and 1997. The new cost accounting method that we developed according to the activity-based costing method was applied in the following four levels; major items of expenditure for the hospital; costs incurred in each department; costs per medical service units; and costs of all the services per case. RESULTS: 1) 158 patients were studied. All the cost figures are adjusted to those in the 1998 fiscal year. The mean length of stay (LOS) of the 158 cases were 52 +/- 16 days. The average charge was 1,835,000 yen, and the average costs was 2,034,000 yen. 2) The per capita ratio of charge to cost (RCC) was 0.90. RCC for medications, procedure treatments, laboratory tests, and medical management/accommodation were 1.04, 1.44, 1.35, and 0.31, respectively. 3) The peason's correlation coefficient between the total costs and LOS was 0.80 (P < 0.001). A high correlation was noted for costs for medical management/accommodation and nursing with LOS (r = 0.98, P < 0.001; r = 0.97, P < 0.001; respectively), while that for cost for operations was low (r = 0.03, P > 0.05). The partial correlation coefficient between the total costs and the total charges with the LOS adjustment was 0.58 (P < 0.001). The coefficient for costs and charges for medications and procedure were high (r = 0.99, P < 0.001; r = 1.00, P < 0.001), while that for medical management/accommodation was low (r = 0.16, P < 0.001). CONCLUSIONS: LOS reflected the cost for room and nursing, but not the resource consumption for medical treatment per case. While the present fee schedules overestimate the costs of medication and laboratory tests, they underestimate those for medical management/accommodation. LOS and charges did not correctly reflect the medical costs per case. PMID- 12772612 TI - [A follow-up study on mental and physical function function among stroke patients: comparisons between admission and discharge status in a rehabilitation specialized hospital]. AB - PURPOSE: To assess results of a 5 year-follow-up, in terms of disability, depression, quality of life, and social network in cases of stroke patients. This report deal with results at admission and discharge (in average, from 2.5 to 6 months of the onset) in a rehabilitation-specialzed hospital. OBJECT AND METHOD: The subjects were 87 stroke patients hospitalized in the Ibaraki Prefectural University Hospital. They were 64 men and 23 women, and their age were between 42 and 79 with an average of 59 years. Systematic evaluation for mental and physical functions was conducted at admission and discharge, and it is intended that the evaluation will be repeated 1, 2, 3 and 5 years from the stroke onset. RESULTS: There were substantial improvements in motor functions, general intelligence, and the status of dementia during the hospitalization. Activities of daily living and degrees of job execution and satisfactory were also improved. The prevalence of depression remained 40%, and the acceptance of limb paralysis or reduced quality of life did not change. Social network scores declined. DISCUSSION: Lack of improvement of depressive status and quality of life, and deterioration social network may increase the rist of major difficulties after discharge of stoke patients. Prevention programs at hospital may be necessary to cope with these potential problems. PMID- 12772613 TI - [Utility of RFLP analysis for tuberculosis case management in Okinawa Prefecture]. AB - SETTING: Routine contact actions for tuberculosis (TB) case management at Public Health Centers (PHCs) in Okinawa Prefecture, Japan. OBJECTIVE: To examine the utility of RFLP analysis in routine contact actions for tuberculosis at PHCs and in the epidemiological surveillance of tuberculosis in Okinawa Prefecture, Japan. DESIGN: An RFLP analysis was conducted for all isolates of TB bacilli cultured from newly registered TB patients in Okinawa Prefecture between April 1996 and September 1997. When an identical band pattern was seen in isolates from two or more patients, the staff of the PHCs discussed the possible epidemiological link based on information collected from the patients by public health nurses at interviews in routine patient service. RESULTS: The main hospitals with TB beds, which diagnose most of the TB patients in the prefecture, and the bacteriological laboratories of the PHCs, submitted TB isolates to the Research Institute of Tuberculosis, Tokyo, for analysis. The prefecture and the PHCs conducted an additional epidemiological investigation based on the analysis results. All these activities were supervised by the Tuberculosis Surveillance Committee of the Prefecture. This organization has proven to work well, so that more than 75% of bacteriologically confirmed TB patients' strains were able be analyzed during this study period. A total of 263 samples recovered from 229 TB patients were collected and analyzed. From these, seventeen clusters have been identified, varying in size from 2 to 10. Five clusters had members that appeared to be family or friends. One cluster appeared to have a common infectious source that had not clearly been detected before the RFLP analysis. However, there was also one large cluster with members that were difficult to connect to a common source of infection, suggesting the existence of an endemic strain causing isolated infections. In many cases, links suspected through the routine contact actions by public health nurses were further supported by the RFLP analysis results. CONCLUSION: RFLP analysis can be a useful tool for TB control activities of the community level, such as in Okinawa Prefecture. First, it gives clues to understanding the epidemiological situation of TB infection in a population. Second, it facilitates detection common infectious sources that might otherwise remain undetected by routine contact actions. Third, it assist public health nurses' activities in routine epidemiological investigations of patients and their contacts in TB case management. PMID- 12772614 TI - [The effects of a practical diabetes prevention training course for public health care providers in Aichi Prefecture]. AB - PURPOSE: Lifestyle intervention for high-risk people is one of the most important issues for reduction of diabetic patients. Public health care providers should update their knowledge and enhance their educational skills. The purpose of this study was to evaluate the effects of a diabetes prevention training course for public health care providers in Aichi Prefecture. METHODS: This course included not only lectures about up-to-date knowledge about diabetes but also many practical aspects such as lifestyle assessment and coaching methods. The participants were made aware of an adequate diet by choosing foods in nutrition sessions, and they experienced walking and aerobic exercise with heart-rate monitors and pedometers in an exercise program. They also learned how to interview and provide health guidance to impaired glucose tolerance (IGT) people through role play. There were 373 participants from 90% of municipal health care centers in Aichi Prefecture. We made inquiries about the effectiveness of this practical training course immediately after the training period and 6-months later for all participants. In follow-up inquiries, we asked if the knowledge obtained in this training course had been useful and had improved their services. RESULTS: The aim and methods were well-accepted by more than 90% of participants. The follow-up examination (6 month later) showed improvement with reference to teaching methods (60%), evaluation of the diabetes prevention service (53%), cooperation with other facilities (48%) and survey on municipal health problems related to diabetes (48%). New methods, which they experienced in this course, were adopted in their own services in many municipalities. Many municipal health care centers started the diabetes individual health promotion program, because the health care providers got aware of importance of diabetes prevention by attending this training course. DISCUSSION: In order to make effective interventions, public health care providers should obtain up-to-date knowledge about diabetics and educational skills. Administrative organ can play important roles in diabetes prevention by training public health care providers and setting up a network between related facilities. PMID- 12772615 TI - [Test-retest variation in the Tokyo Metropolitan Institute of Gerontology Index of Competence in community-dwelling older people independent in daily living toward individual assessment of functional capacity]. AB - OBJECTIVES: To assess individual variation in the functional capacity of community-dwelling older people who are almost independent in daily living, using the Tokyo Metropolitan Institute of Gerontology Index of Competence (TMIG-Index of Competence). METHODS: Out of 74 elderly outpatients showing a total score of TMIG-Index of Competence above 10 points at baseline, 61 who remained stable during subsequent two months in terms of medical and social aspects were assessed for their functional capacity with the test-retest method using the TMIG-Index of Competence. We assessed individual variation between the tests of one month interval with correspondence rate. RESULTS: The ranges for score differences within which correspondence rates of 95% and over applied were as follows: within 1 point for the total score of the Index (95.1%), 0 point in the Instrumental Self-Maintenance subscale (IADL) (95.1%), and 1 point in the Intellectual Activity subscale (Intellectual Activity) (98.4%) and the Social Role subscale (Social Role) (98.3%). CONCLUSIONS: The variation of 1 point for the total score of the Index, and subscales of Intellectual Activity and Social Role was regarded as a possible measurement error. In other words, variations of 2 points and over for total score, subscales of Intellectual Activity and Social Role, and variations of 1 point and over for IADL should not be ignored in screening of functional capacity among older people. PMID- 12772616 TI - Qualitative versus quantitative methodologies: and never the twain shall meet? PMID- 12772617 TI - Development of a family assessment instrument for transcultural use. AB - PURPOSE: The article describes challenges related to the transfer of a family assessment instrument, the Assessment of Strategies in Families-Effectiveness (ASF-E), and its theoretical basis to another culture. DESIGN: Instrument development encompassed translation and discussion of theoretical and cultural relevance of the items. Patients from two clinics in Tampere, Finland (N = 196) completed the ASF. Psychometric tests involved factor and reliability analysis. RESULTS: The theoretical constructs of stability, growth, control and connectedness/spirituality were represented in the factor structure, and reliability ranged from .65 to .83. DISCUSSION: There were cultural differences in the interpretation of certain items. Characteristics of Finnish culture with impact on instrument validity were homogeneity and family emphasis on conflict resolution. Insights contributed to further instrument development and broader understanding of culture. PMID- 12772618 TI - A discussion of chain referral as a method of sampling hard-to-reach populations. AB - Nursing research often requires inquiry into sensitive topics that involve hidden or hard- to reach populations. However, identifying and sampling these populations for research purposes is often fraught with difficulties. Barriers include society's lack of tolerance of diverse groups, social stigma, concern for issues of confidentiality, and fear of exposure because of possible threats to security. Chain referral sampling techniques are proposed to minimize bias while maintaining privacy and confidentiality. Techniques of chain referral sampling are detailed for use in researching sensitive topics and hidden populations. When carefully planned and executed, this sampling design offers transcultural nurse researchers a reasonable method for accessing and studying special populations that are particularly hard-to-reach. PMID- 12772619 TI - Physical activity beliefs, behaviors, and cardiovascular fitness of Mexican immigrant women. AB - This descriptive study examined physical activity beliefs, behaviors, and cardiovascular fitness of a convenience sample of 51 married Mexican immigrant women (MIW) with children in Northern California. Standardized tools/procedures were used to measure the variables of interest and to determine cardiovascular fitness levels. The majority (78%) was not involved in regular physical activity and had below average or poor cardiovascular fitness (76%). However, 93% had a positive attitude, were well-informed of the benefits, and perceived physical activity to be a health-promoting behavior. Cultural values and beliefs about physical activity, gender roles, and social and physiological factors were described as barriers to women's intention to engage in physical activity. PMID- 12772620 TI - Come bien, camina y no se preocupe--eat right, walk, and do not worry: selective biculturalism during pregnancy in a Mexican American community. AB - Mexican American childbearing women appear to offer a healthy model for pregnancy. However, statistics suggest that they may be at increased risk for poor birth outcome as they acculturate to a U.S. lifestyle. An ethnographic study in Watsonville, California, examined the influence of acculturation on pregnancy beliefs and practices of 29 Mexican American childbearing women. Data from formal semi-structured interviews were submitted to content analysis. During pregnancy, women balanced well-documented, traditional Mexican cultural beliefs with the individualistic beliefs common to Anglo-Americans. Selective biculturalism emerged as a protective approach to stress reduction and health promotion. Stress reduction interventions as part of routine prenatal care have potential benefit for all pregnant women. Future research on cultural barriers to family-based social support during pregnancy is needed. PMID- 12772621 TI - Multiple role stress and patterns of coping of Egyptian women in clerical jobs. AB - Egyptian women are increasingly becoming a significant work force; however, many of these women continue to occupy low-income jobs. In the study reported here, 190 women in Egypt employed in clerical jobs were asked about the satisfaction and stress they experience in their work and maternal roles. They were also asked about their coping approaches and the demands in their daily lives. The Women's Role Interview Protocol was used to collect data. The results were analyzed within the symbolism of the Arab/Egyptian language using thematic and content analysis. A prevailing theme is the interconnectedness among all women's roles when describing their stress, satisfaction, and coping. Another major theme that transcends all roles is the perception of inequality and how it affects their daily lives. Emergent stressors were grouped under employment role stress, maternal role stress, marital role stress, and relational role stress. Women coped through learning to be self-reliant and by using cognitive and emotion focused coping approaches. Women's stress was embedded in inequality in gender roles, and the women are empowered to cope through relying on endurance and outliving conflict. PMID- 12772622 TI - Nursing in a global community: a study abroad program. AB - Nursing in a global community is a study abroad program at Georgia Southern University School of Nursing. The program hosted at the University of Cape Coast in Ghana, West Africa, provides opportunity for undergraduate and graduate health profession students to experience nursing, health care service and delivery in a developing country. Description of the program and benefits of study abroad programs are discussed as well as a student perspective on the transcutural experience in Ghana. PMID- 12772623 TI - A look into the Amish culture: what should we learn? AB - It is important to understand the Amish culture in order to provide appropriate, acceptable, and accessible health care to this culturally diverse group. A case study pertaining to the care of a dying elderly Amish woman living in a rural Amish community is examined. This allows for exploration into the world of the Amish community in greater detail. Their overall beliefs, values, and behavior are discussed as well as how their lifestyle affects their health care decisions, access to health care, and reimbursement of services. Nurse practitioners can offer culturally sensitive and appropriate health care to the Amish population by recognizing important cultural values that have survived for more than three hundred years. PMID- 12772624 TI - Diversity in care values and expressions among Turkish family caregivers and Dutch community nurses in The Netherlands. AB - Aside from being a world harbor, Rotterdam is known for its large numbers of migrant groups. The Turkish community comprises one of the largest groups of migrants. This article reflects on the findings of an ethnonursing research, Transcultural Care for the Elderly at Home, which explored diversity and universality in care values and meanings relevant to care of the elderly at home between Turkish lay caregivers and Dutch professional nurses in the community. It compares the caring values and patterns of these two groups of caregivers within the cultural context of Turkish families in Rotterdam. Based on the comparative findings of the research, recommendations for implementing culturally congruent nursing care for Turkish elders at home are presented. PMID- 12772625 TI - Founder's focus: transcultural nursing care makes a big outcome difference. PMID- 12772626 TI - Simplifying periodontal care. PMID- 12772627 TI - Getting back to basics. Part 1: An ounce of prevention is worth 1.2 parts per million of fluoride. PMID- 12772628 TI - Single-tooth implants in the maxillary incisor region with immediate provisionalization: 2-year prospective study. AB - Osseointegrated implants have been increasingly used for aesthetic, predictable restorative treatment. This study presents the 2-year postoperative results of patients treated with immediate, single, tapered implants in the maxillary incisor region and the simultaneous placement of provisional implant-supported crowns. Of the total of 92 implants placed, 6 were lost. Eighty-six implants were reevaluated and judged to have no signs of mobility, peri-implant inflammation, or adverse reactions. When used in selected cases, this technique facilitated maintenance of the gingival architecture adjacent to immediate transalveolar implants. PMID- 12772629 TI - Technical strategy: maintaining open patient communication for anterior restorative success. PMID- 12772630 TI - CT scans for endodontic diagnosis. PMID- 12772631 TI - The role of the implant housing in obtaining aesthetics: generation of peri implant gingivae and papillae--Part 1. AB - The use of osseointegrated implants provides predictable, long-term aesthetics, providing that the implant housing and peri-implant gingivae are adequately maintained. Restoration-driven implant placement must, therefore, result in the development of harmonious peri-implant soft tissue contour as compared to the adjacent natural teeth. This article analyzes the implant housing features that influence aesthetics in implant therapy. Case presentations that include surgical and restorative innovations are described to illustrate methods used to achieve aesthetics with single-tooth implant restorations. PMID- 12772632 TI - Is there a better way to select dental laboratories? PMID- 12772633 TI - Lasers in the dental office: treatment considerations for hard and soft tissue contouring. PMID- 12772634 TI - Primary stability and osseointegration: preliminary clinical results with a tapered diminishing-thread implant. AB - Due to their success and predictability, osseointegrated implants are used with increasing frequency for the prosthetic restoration of edentulous patients. Analysis of implant failures, however, enables clinicians to improve the clinical results achieved with this modality. While the literature discusses varying etiological factors (e.g., poor surgical technique, patient immune factors, insufficient bone quality or quantity), this article addresses primary stability as a predicator of implant success. The article also demonstrates the use of an anatomically shaped implant system that may improve primary stability in common clinical scenarios. PMID- 12772635 TI - Gingival reconstruction with an onlay-interpositional graft following excision of a pyogenic granuloma: description of a surgical approach. PMID- 12772636 TI - Cures from the cosmos. PMID- 12772637 TI - Bohn voyage. PMID- 12772638 TI - Unsettling practices. PMID- 12772639 TI - Buying health care off the rack. PMID- 12772640 TI - Assessing your operations. PMID- 12772641 TI - Community preceptors' attitudes toward and practices of complementary and alternative medicine: a Texas survey. AB - To assess attitudes toward and practices of complementary and alternative medicine (CAM) in Texas, a survey was developed and sent to 143 community preceptor physicians in September 2001. The results of the survey represent what the physicians of Texas believe about CAM as a practice and about the relationship of CAM to the medical community. Results indicate that the concept of CAM is relatively well accepted, continuing education is enthusiastically accepted, and support for medical school education is strong; however, physicians also report concerns over the attitudes of their colleagues toward CAM as well as a wide range of concerns about CAM therapy in general. The physicians verify that patients seem to expect them to be knowledgeable about CAM therapies and note some frustrations with the lack of opportunity for fact-based continuing education. PMID- 12772642 TI - Oral complications of cancer therapy. AB - Oral complications of cancer therapy include a multitude of conditions that compromise a patient's quality of life and can be potentially life threatening. Severe oral pain, infection, and bleeding can result. Often, these oral complications cause postponement of cancer therapy and hospitalization for management. Oral intervention before and after radiation, chemotherapy, and bone marrow and stem cell transplantation can greatly reduce these complications. A close working relationship between oncologists and dental professionals should be established to successfully manage patients undergoing cancer treatment. PMID- 12772643 TI - [A positive vision]. PMID- 12772644 TI - [Microbicides and women]. PMID- 12772645 TI - AIDSWatch 2003 brings advocates to D.C. PMID- 12772646 TI - CDC unveils HIV testing strategy. PMID- 12772647 TI - Corporate responsibility. Shareholders challenge Pepsi over AIDS response in Africa. PMID- 12772648 TI - Medical center ordered to repay Medicaid payments to state. PMID- 12772649 TI - Advocates say CDC's new testing initiatives are off target. PMID- 12772650 TI - HIV/AIDS prevention programs to lose federal funding. PMID- 12772651 TI - Politics. Grant applicants told to avoid 'controversial' language. PMID- 12772652 TI - Malpractice. Conviction upheld for doctor who treated AIDS with fen/phen. PMID- 12772653 TI - Disability. Supreme Court decision further erodes ADA protections. PMID- 12772654 TI - HIV-positive prisoner loses excessive force claim against officers. PMID- 12772655 TI - Budget, policy woes continue to squeeze HIV/AIDS programs. PMID- 12772656 TI - First amendment. Government officials denied immunity in retaliation case. PMID- 12772657 TI - [Music heals us. Report on Musica Against Drugs]. PMID- 12772658 TI - [Creating music, creating conscience]. PMID- 12772659 TI - [How the immune system protects us against disease]. PMID- 12772661 TI - [Tenofovir (Viread)]. PMID- 12772660 TI - [Should I worry if my cholesterol is high?]. PMID- 12772662 TI - Mitochondrial toxicity and lactic acidosis. AB - Mitochondrial toxicity and one of its symptoms called lactic acidosis have been highlighted recently as a previously undiagnosed side effect of anti-HIV drugs. Some researchers believe that mitochondrial toxicity contributes to the fat redistribution (lipodystrophy) associated with anti-HIV therapy. Although much more research is needed to fully understand this issue, this article explores the current thoughts about the connections among mitochondrial toxicity, lactic acidosis and lipodystrophy. PMID- 12772663 TI - Resistance testing comes of age. AB - The Antiviral Drugs Advisory Committee (ADAC) to the Food and Drug Administration (FDA) recently held a two-day meeting to discuss HIV drug resistance testing. The discussion centered on currently available results from drug resistance tests and not on any one specific test or drug. For more information on HIV drug resistance and resistance testing, call Project Inform's Hotline and ask for the Geno- & Phenotypic Resistance Tests Quick Sheet. PMID- 12772664 TI - New information on opportunistic infections and co-infections. AB - Many reports have shown fewer opportunistic infections (OIs) and deaths since the wide-scale availability and use of potent anti-HIV therapy. Also, there have been many reports of people stopping preventive therapies for OIs. Now several studies further confirm that it may be safe for some people to stop preventive therapies when their CD4+ cell count increases are sustained over time. In addition, ongoing research into hepatitis co-infection (with HIV) is beginning to show progress. PMID- 12772665 TI - Treating and preventing fungal infections naturally. AB - There is a strong connection between what you eat and the health of your immune system. Nevertheless, nutritional approaches to prevent and treat conditions like candidiasis (fungal, yeast infections) are complicated and controversial. While there isn't a magic recipe that prevents or treats yeast infections in everyone, following some basic guidelines may lower the risk of yeast becoming a problem. PMID- 12772666 TI - Candidiasis update. AB - Sustained CD4+ cell count increases as a result of potent anti-HIV therapy have been credited with a steep drop in many HIV-related conditions. Indeed, the most recent guidelines for preventing opportunistic infections (OIs) reflect the growing practice of stopping preventive and maintenance therapies. While the risk of developing OIs or having them reappear still exists only after therapy fails, the possibility of stopping OI therapy after achieving an improved immune response is promising for many. Unfortunately, despite success with many OIs, declines in the rate of the fungal infection candidiasis have not been observed. PMID- 12772667 TI - 1999 National Conference on Women and HIV/AIDS. AB - The 1999 National Conference on Women and HIV/AIDS, held last October in Los Angeles, convened its largest number of participants. Over 2,000 researchers, clinicians, policymakers, AIDS activists and HIV-positive women came to discuss new treatment, behavioral and political issues affecting women. The strong and diverse turnout provided a lively and sometimes conflicting forum that reflected the growing visibility of women's issues. Following are selected highlights from the conference. For more on the 1999 Conference, call Project Inform's Hotline and ask for WISE Words #5, which fully covers the conference. PMID- 12772668 TI - Summary of the eighth Immune Restoration Think Tank. AB - Project Inform hosted the eighth meeting of the Immune Restoration Think Tank: The Dobson Project (IRTT), October 8-10, 1999 in Chicago. Researchers from around the world, in partnership with community activists, a convened to review data from ongoing projects, previously inspired through the Think Tank effort, and discuss new strategies and areas of focus for immune reconstitution research in people with advanced stage AIDS. PMID- 12772669 TI - Immune-based therapies in brief. AB - Immune-based therapy is considered by many to be the "holy grail" of AIDS research. Most successful therapies to date have been anti-HIV drugs that suppress reproduction of HIV, the virus that causes AIDS. Success, however, usually comes at the price of many side effects. Researchers and patients alike have longed for therapy that doesn't so much try to wage war against the virus, but rather strengthens the weakening immune response associated with AIDS. The hope is that such an approach might be more natural or less toxic. Although health food stores offer an abundance of products that claim to "boost" or "strengthen" immunity, those are mostly just advertising claims. Helping the immune system in its battle against HIV is far more complex than simply "boosting" anything. PMID- 12772670 TI - Post exposure prevention. AB - There has been growing interest in the use of Post Exposure Prevention (PEP) as a means of preventing HIV infection. PEP involves a person beginning anti-HIV therapy within hours of suspected HIV exposure, with a goal of blocking the establishment of HIV infection. PMID- 12772671 TI - The global crises: AIDS in Tanzania. AB - More than 90% of people living with HIV/AIDS are in the developing world, most of whom live in Africa, followed by Asia and Latin America. In an effort to increase awareness of the HIV epidemic in these regions, PI Perspective will feature short profiles of the epidemic in some of these countries. In this issue, PI Perspective looks at the United Republic of Tanzania. PMID- 12772672 TI - Global strategies for mother-to-child HIV transmission prevention. PMID- 12772673 TI - Reznik named to PACHA. PMID- 12772674 TI - Will sneezing be a felony in Georgia? PMID- 12772675 TI - "The personal is STILL the political". PMID- 12772676 TI - Women and disclosure. PMID- 12772677 TI - Spotlight on SisterLove. PMID- 12772678 TI - PACHA's growing pains. PMID- 12772679 TI - Women's health issues from the 2003 retrovirus conference. PMID- 12772680 TI - Preventing and treating vaginal yeast infections. PMID- 12772681 TI - Burns. What to do right away. PMID- 12772682 TI - Health tips. Leaves of three, leave them be. PMID- 12772684 TI - Behavior therapy and urge incontinence. PMID- 12772685 TI - Being a cancer survivor. Rediscovering 'normal'. PMID- 12772683 TI - New treatment options for atopic dermatitis. PMID- 12772686 TI - Staying mentally sharp. Can you train your brain? PMID- 12772687 TI - Pleurisy. When breathing hurts. PMID- 12772688 TI - I get occasional diarrhea. Is there anything nonprescription that's particularly effective for that? PMID- 12772689 TI - I have a little sore on the side of my tongue. It's tender to the touch and has been there almost 3 weeks. Should I see a doctor? PMID- 12772690 TI - Protecting children in a radiation disaster. PMID- 12772691 TI - Echinacea for the common cold. PMID- 12772692 TI - Bullying and violence. PMID- 12772693 TI - Child safety. Too many are too small for a seatbelt. PMID- 12772694 TI - Toilet training: does an early start help? PMID- 12772696 TI - Product recalls. Recalls: peanut allergy alert...party poppers sold at Wal Mart...and Dollar Tree Stores plush bears and snowman dolls. PMID- 12772695 TI - Resource: where to get the latest information on SARS. PMID- 12772697 TI - Special issue: gravitational biology. PMID- 12772698 TI - In vitro fertilization may increase the risk of Beckwith-Wiedemann syndrome related to the abnormal imprinting of the KCN1OT gene. PMID- 12772699 TI - To trust or not to trust an idiosyncratic mitochondrial data set. PMID- 12772701 TI - Improved specificity of Y-STR typing in DNA mixture samples. AB - Y-STR loci are beneficial for the analysis of forensic samples especially in sexual assault cases or other situations where mixtures of male and female cells are present. However, the amplification of Y-chromosomal STRs is also known to result in the formation of artefactual amplification products, mainly due to insufficient PCR specificity. This is a major drawback of the method, as the sensitivity as well as the correct Y-STR interpretation are affected. In our study, the addition of a PCR enhancer to the reaction master-mix resulted in a significant increase of specificity of Y-STR typing. This was clearly demonstrated by a loss of artefactual signal with increasing enhancer concentration, while the peak heights of the Y-STR alleles were not significantly affected by the enhancer. Mixtures of up to 1:500 (200 pg male and 100 ng female DNA) gave correct Y-STR profiles when the PCR enhancer was added to the reaction, while artefactual amplification succeeded over Y-specific amplification when no PCR enhancer was present. PMID- 12772700 TI - A multicolor FISH assay does not detect DUP25 in control individuals or in reported positive control cells. PMID- 12772703 TI - Health care workforce issues in Massachusetts. PMID- 12772704 TI - Medical errors and patient safety in Massachusetts: what is the role of the Commonwealth? PMID- 12772705 TI - Dynamic contrast-enhanced MRI of the prostate: is this the way to proceed for characterization of prostatic carcinoma? PMID- 12772706 TI - Eliminating racial and ethnic health disparities: opportunities and challenges for the Commonwealth. PMID- 12772707 TI - Richard H. Ward, Ph.D. (June 7, 1943-February 14, 2003): wild ride of the Valkyries. PMID- 12772708 TI - Acute right upper quadrant pain and anaemia. Ruptured hepatic adenoma. PMID- 12772709 TI - Multiple retained acupuncture needle fragments. PMID- 12772711 TI - An unusual cause of knee pain (2003:5a). PMID- 12772710 TI - En-plaque tuberculomas of tentorium in a pregnant woman: follow-up with MRI(2003:2b). PMID- 12772712 TI - Antacid powders. PMID- 12772713 TI - The pseudohumerus. Aneurysmal dilatation of axillary and brachial arteries secondary to a radiocephalic fistula. PMID- 12772714 TI - Spotlight on research: psychosocial intervention for newly diagnosed cancer patients. PMID- 12772715 TI - The quota: "an equally serious problem" for us all. AB - The author explores the question--was a quota applied to the admission of women to the medical school of the University of Toronto? She uses a variety of sources, including class photographs, archives, statistics, personal recollections, and oral history. The existence of a quota from 1944 to 1968 is inferred from statistical patterns and confirmed by a surprising source. In closing, the historiographic implications of this project for research on other politically charged topics are considered. PMID- 12772717 TI - Current world literature. Epidemiology and prevention. PMID- 12772716 TI - Current world literature. Renal immunology and pathology. PMID- 12772718 TI - [The 103rd annual meeting of the Japan surgical Society. Hokkaido, Japan. June 4 6,2003. Abstracts]. PMID- 12772719 TI - [Transition in the articles published in the Japanese Journal of Public Health (from Volumes 1 to 49)]. PMID- 12772720 TI - Role of oxygen in corneal epithelial homeostasis during extended contact lens wear. PMID- 12772721 TI - Can variability in corneal metabolism explain the variability in corneal swelling? AB - PURPOSE: To examine if the variability in hypoxic corneal swelling in non-contact lens wearing and contact lens wearing groups can be explained by the variability in corneal metabolic activity or endothelial function. METHODS: Corneal thickness was measured by Orbscan before and after 2 hr of closed-eye hydrogel lens wear. De-swelling rates (percentage recovery per hour) were determined by continued corneal thickness measurements until open-eye steady-state thickness was reached. Corneal oxygen consumption was estimated by measuring tear oxygen tension while wearing hydrogels of known oxygen transmissibility. The change in stromal pH because of hypoxia was measured by ratio fluorescence of stromal fluorescein. RESULTS: Corneal swelling was inversely correlated with open- or closed-eye tear oxygen tension while wearing the test hydrogels. Corneal swelling was directly correlated with the change in stromal pH during hypoxic lens wear. Unexpectedly, corneal swelling was also inversely correlated with the percent recovery per hour. These parameters could not explain all the swelling variability. The contact lens wearing group showed significantly less swelling and change in stromal pH with the same lenses. However, percent recovery per hour and tear oxygen tension was the same. CONCLUSION: Corneal swelling is associated with corneal metabolic activity and endothelial function; however, other factors must also be involved. Contact lens wearers do not adapt by modifying oxygen consumption but probably by using glucose more efficiently. PMID- 12772722 TI - Innate defense of the ocular surface. AB - PURPOSE: The mucosal surface of the eye interfaces with the environment and can detect the presence of pathogens and irritants. The mucosal cells respond by up regulating genes whose products either directly attack the contaminant or recruit leukocytes through the release of cytokines. Mucin, an epithelial glycoprotein, traps contaminants and aids in their removal through normal tear clearance mechanisms. METHODS AND RESULTS: Using Northern analysis and reverse transcription-polymerase chain reaction, we have found that MUC1, a membrane bound ocular surface mucin, is up-regulated following exposure to the products of both bacteria and activated lymphocytes. Consistent with this, MUC1 gene expression is also up-regulated in corneal epithelial cells exposed to precorneal tear film collected from two rigid contact lens wearers. CONCLUSION: The changes in MUC1 gene expression during exposure to potential ocular irritants suggest that analysis of the molecular mechanisms mediating these changes may reveal control points amenable to drug therapy. PMID- 12772723 TI - Effects of daily and overnight wear of hyper-oxygen transmissible rigid and silicone hydrogel lenses on bacterial binding to the corneal epithelium: 13-month clinical trials. AB - PURPOSE: For 14 years, the U. S. Food and Drug Administration (FDA) and eye care practitioners have strongly discouraged patients from sleeping in contact lenses. In the past 9 months however, the FDA has approved three new hyper-oxygen transmissible lenses for up to 30-night extended wear. Is this a great advance or another clinical triumph of hope over experience? What should the public know? What should patients do? METHODS: Our research group has studied all three new lenses in prospective, randomized, masked, parallel clinical trials in a single center. As an outcome measure, we looked at whether lens wear caused more bacterial binding to surface corneal cells. RESULTS: Compared with conventional lens use, the new lenses caused no or only small increases in bacterial binding in either daily or extended wear. Furthermore, the increases seen stratified with known infection risks by both lens type (hard or soft) and wearing schedule. Indeed, early epidemiologic reports indicate that this new generation of lenses may reduce lens-related microbial infection risks by 10- to 40-fold. CONCLUSION: This represents a true clinical paradigm shift of the first magnitude in safety of contact lens wear. Taken together, this data suggest that patients will soon replace their current, conventional lenses with this new generation of materials for any schedule of wear. PMID- 12772724 TI - Oxygen permeability of the hypertransmissible contact lenses. AB - PURPOSE: Oxygen permeability (Dk) was assayed in a masked fashion using the polarographic method for three materials found capable of hypertransmissibility as contact lenses: tisilfocon A, balafilcon A, and lotrafilcon A. METHODS: Two sets of tisilfocon rigid lenses were tested in six thicknesses and two sets in nine thicknesses for the hydrogels. The two sets of tisilfocon were from two different lots and of lotrafilcon were from different base curve radii (8.6 and 8.4 mm). The six test sets were measured with the inclusion of rigid reference materials for the purpose of simultaneous calibration. RESULTS: Oxygen permeability (in barrers) of the two lots of tisilfocon A was 159.6 (95% confidence interval [CI], 146.3-175.5) and 175.1 (95% CI, 154.9-201.6). The Dk values for balafilcon A were 111.3 (95% CI, 103.6-120.3) and 102.0 (95% CI, 90.5 116.7), and the Dk values for lotrafilcon A were 176.1 (95% CI, 165.1-188.7) and 190.2 (95% CI, 177.5-205.0) The coefficients of determination (R2) of the corrected resistance versus thickness plots were 0.98 or more. Paired-comparison t tests were performed between the sets of each test material paired by thickness. These revealed the oxygen transmissibilities (Dk/L) of the lenses made from the two lots of tisilfocon A to be significantly different (t = 2.62; P < 0.047; df = 5) and lenses of the two base curve radii made of lotrafilcon A to be significantly different (t = 4.42; P < 0.002; df = 8). These differences were attributed to a material difference in Dk between lots of tisilfocon A and a thickness difference between lens designs of lotrafilcon A. CONCLUSION: Oxygen permeability of tisilfocon was found consistent with that claimed by the manufacturer (163 barrers). Dk of balafilcon was found marginally greater than claimed (99 barrers) and of lotrafilcon, greater than claimed (140 barrers). Preliminary Dk/L values of two of the three test materials were statistically different between Sets 1 and 2, yet they were inside the American National Standards Institute Z80.20-1998 tolerance (+/-20%). Resistance-versus-thickness plots were linear up to a Dk of at least 190 barrers. The Dk/L values over the central zone of minus and low-plus contact lenses made of these materials are predicted to decrease on the upper asymptotic portion of the Dk/L versus equivalent oxygen percentage curve. Hence, the physiologic effects of oxygen variability from lot-to-lot, lens-to-lens, and site-to-site on the same lens will be minimized. PMID- 12772725 TI - Clinical signs of hypoxia with high-Dk soft lens extended wear: is the cornea convinced? AB - PURPOSE: To assess the effectiveness of high-Dk soft contact lenses with oxygen transmissibility (Dk/L) beyond the critical level required to avoid corneal edema during overnight wear. METHODS: The most up-to-date data available on clinical signs of hypoxia with high-Dk contact lenses is reviewed. RESULTS: Chronic corneal edema associated with hypoxia is responsible for the development of large numbers of microcysts, limbal hyperemia, neovascularization, and small increases in myopia. Silicone hydrogel lenses worn continuously for up to 30 nights prevent corneal edema during overnight wear and do not induce a microcyst response. Long term clinical trials indicate the mean level of limbal redness for patients wearing high-Dk lenses during continuous wear are equivalent to nonlens wearers. No changes in refractive error are associated with continuous wear of high-Dk lenses. CONCLUSION: High-Dk silicone hydrogel lenses can be worn for up to 3 years with virtual elimination of the hypoxic consequences observed with low-Dk lenses made from conventional lens materials. PMID- 12772726 TI - Continuous wear contact lens surface chemistry and wearability. AB - Continuous wear (CW) contact lenses are defined as lenses composed of hydrogel polymers containing elements other than carbon, hydrogen, oxygen, and nitrogen that enhance oxygen permeability to an extent greater than water alone. Those elements are silicon and fluorine. Silicon is incorporated as siloxanes, and fluorine is used as fluoroalkyl. Despite the water present in CW lenses, they are not wearable without surface modification because of the tendency of siloxanes and fluoroakyls to move in the soft polymers, orient, and become enriched at the surface. Various methods of surface modification are discussed, with emphasis on the plasma technologies used by the two commercial CW lens products, Focus Night & Day and PureVision. Speculation about future directions in surface chemistry are also presented. PMID- 12772727 TI - The effects of ocular and lens parameters on the postlens tear thickness. AB - PURPOSE: To assess the effects of soft contact lens base curve radius (BCR), sex, ethnicity, central corneal curvatures, and vertical palpebral aperture size (PAS) on the postlens tear thickness (PLTT). METHODS: The PLTT was measured using optical pachometry on 114 experienced lens wearers who were fitted with lathe-cut soft lenses (Alden 47, polymacon, 35.5% H2O, -2.00 diopter, and 14.0 mm). Each subject was randomly allocated to one lens group receiving a BCR of 7.9, 8.3, or 8.7 mm. Pachometry measurements were taken at 30 min after lens insertion. Vertical PAS and keratometry readings were measured for 94 of the 114 subjects. RESULTS: The mean (95% confidence interval) PLTT was 15.7 microm (13.2-18.0 microm), 12.8 microm (10.9-14.7 microm), and 12.1 microm (10.2-14.0 microm) for the 7.9-mm, 8.3-mm, and 8.7-mm BCR groups, respectively. The differences in PLTT among the three BCR groups was significant (analysis of variance F-test; P=0.039). Post hoc testing using the Tukey honestly significant difference statistic showed that only the two extreme BCR groups (7.9 mm and 8.7 mm) were significantly different. Sex had no effect on the PLTT; however, the PLTT was significantly thinner for the Asian compared with non-Asian eyes (P=0.0001). The Asian PLTT did not vary with different soft lens BCRs. The non-Asian PLTT was thicker with lenses of the steep BCR compared with the flat BCR. CONCLUSION: These results show that the base curve radius of a soft contact lens and several ocular characteristics can affect the thickness of the postlens tear film. PMID- 12772728 TI - Contact lens chemistry and giant papillary conjunctivitis. AB - OBJECTIVE: To review the relationship between contact lens chemistry and the occurrence of giant papillary conjunctivitis (GPC). DESIGN: A review of the literature. RESULTS: GPC can occur with any type of contact lenses. The coating on the lens and the contact lens trauma to the conjunctiva are probable factors. Changing the polymer of the contact lens that a patient with GPC wears can decrease the chance of the condition recurring. Also, replacing a soft contact lens at intervals of less than 3 weeks, rather than 4 or more weeks, significantly reduces the chance of developing GPC. GPC can occur with high Dk silicone contact lenses. Two forms of GPC have been reported: a generalized form similar to that seen with conventional soft contact lenses and a localized form in which the papillae are confined to one or two areas of the tarsal conjunctiva, near the lid margin. CONCLUSION: Contact lens chemistry plays a role in the development of GPC; however, other factors such as edge design, surface properties, fitting characteristics, and replacement cycle are also important variables in the pathophysiology of GPC. PMID- 12772729 TI - Surface treatment or material characteristic: the reason for the high level of Acanthamoeba attachment to silicone hydrogel contact lenses. AB - PURPOSE: To determine the reason for the high level of attachment of Acanthamebic to silicone hydrogel (SH) contact lenses. The effect surface treatment has on attachment is determined using silicone elastomer (SE) lenses. METHODS: All test lenses were unworn. SH (PureVision), conventional hydrogel (Acuvue), treated SE (Silsoft), and untreated SE (Silsoft) lens quarters were incubated for 90 min with plate-cultured Acanthamoeba castellanii trophozoites. After incubation and rinsing, the trophozoites attached to one surface of each quarter were counted by direct light microscopy. Sixteen replicates were performed for each lens type. Logarithmic transformation of data allowed the use of parametric analysis of variance. RESULTS: Lens polymer had a significant effect on attachment (P<0.001), with higher numbers of trophozoites attaching to the SH and SE (treated and untreated) lenses as compared with the conventional hydrogel. No significant difference in attachment was detected between the SH and SE (treated and untreated) lenses. CONCLUSION: Acanthamoeba attachment to the SH lens was significantly greater than to the conventional hydrogel. The similarity in attachment to surface-treated and non-surface-treated SE lenses suggests that the increased attachment found with the SH lens may be an inherent characteristic of the polymer rather than an effect of the surface treatment procedure. It is possible that SH lenses are at greater risk of promoting Acanthamoeba infection if exposed to the organism because of the enhanced attachment characteristic of this new material. PMID- 12772730 TI - Potential for transmission of prion disease by contact lenses: an assessment of risk. AB - Prions are small proteinaceous infectious agents known to cause central nervous system infections in both animals and humans. Interest in the pathogenesis of these diseases has grown since the emergence of bovine spongiform encephalopathy (BSE or "mad cow disease") in the United Kingdom and several other countries in Europe. Ingestion of meat products from animals infected with BSE has resulted in transmission of the disease to humans as a variant form of Creutzfeldt-Jakob Disease (vCJD). CJD has a long asymptomatic incubation period, is untreatable, and universally fatal. Hence concern has arisen over other possible routes of disease transmission. Because it is known that prions are found in low levels in the corneas of animals with experimentally induced prion disease, and that a case of human CJD transmission by corneal transplantation has occurred, the question of possible prion transmission through the reuse of diagnostic fitting of contact lenses has surfaced. This article reviews prion diseases of animals and humans, and the data regarding the presence and location of prions in the eye. Issues inherent in the question of corneal and contact lens transmission are discussed. Although some key information has yet to be derived, it appears that the chance of obtaining prion disease through contact lens use is negligible. PMID- 12772731 TI - Orthokeratology and corneal refractive therapy: a review and recent findings. AB - PURPOSE: To review the past and current literature and present recent findings on orthokeratology and corneal refractive therapy. METHODS: Various articles on contact lens corneal reshaping were analyzed. Common clinical procedures and interference measurement of tear-film thickness were also used. RESULTS: Although the numbers of patients tested to date do not allow conclusions of great certainty, based on a review of the current literature, our recent study of 60 patients, and the Food and Drug Administration approval of overnight contact lens corneal refractive therapy, there is a low incidence of complications, and unaided visual acuity of 20/20 in the morning is possible in most (74%) successful cases. Refractive error change of 2.25diopter (D) +/- 1.00D is common. A presumed iron ring may appear in some patients in the midperipheral corneal epithelium. CONCLUSION: Overnight orthokeratology and corneal refractive therapy with modern design reverse-return zone lenses in high-Dk rigid gas-permeable contact lens materials is an option for transient vision correction for some myopic patients. PMID- 12772732 TI - Retrospective case series of therapeutic applications of a lotrafilcon A silicone hydrogel soft contact lens. AB - PURPOSE: To report a series of consecutive cases for which a lotrafilcon A silicone hydrogel soft contact lens was used in therapeutic applications. METHODS: Three practitioners in Spain, Denmark, and Germany supplied 41 consecutive case reports on 39 patients for whom a lotrafilcon A silicone hydrogel soft contact lens was used in therapeutic applications for erosion or recurrent erosion, bullous keratopathy, corneal edema, corneal dystrophy, neurotrophic corneal ulcer, entropion, and after corneal surgeries. Practitioners reported their judgements of pain relief, change in corneal signs by slitlamp evaluation, additional complications, and success of therapeutic lens treatment. Twenty cases that had completed treatment and 21 that remained under ongoing treatment were reported. RESULTS: Complete pain relief was reported for 78% of cases with 94% reporting pain relief of 50% or more. Corneal signs were restored to normal for 74% of cases with 100% showing at least partial improvement. No additional complications were reported in 83% of cases. Therapeutic contact lens application was judged fully successful for 71% of cases with 93% showing at least partial success. CONCLUSION: The high-Dk lotrafilcon A lens met or exceeded the outcomes reported from historic reports of therapeutic applications of low-Dk soft contact lenses. Whereas the case reporting conditions may vary among the historic studies and from the current study, there is the indication that success may be improved with the lotrafilcon A high-Dk lens. PMID- 12772734 TI - The causes of and cures for contact lens-induced peripheral ulcer. AB - PURPOSE: Contact lens-induced peripheral ulceration (CLPU) is a relatively common adverse response associated with wearing hydrogel lenses, especially on an extended wear schedule. Bacteriologic examination of lenses at the time of an event has demonstrated an association with Staphylococci spp. We sought to investigate the causes of CLPU in a rabbit model of contact lens wear. METHODS: Rabbits wore contact lenses for a period of 24 hr in the presence or absence of bacteria or in presence or absence of epithelial scratches made in the periphery of the cornea before lens wear. Bacteria tested were a strain of Staphylococcus aureus or a strain of Staphylococcus epidermidis isolated from human CLPUs. Rabbits were also challenged with S. aureus in the presence of an epithelial defect and in the absence of a contact lens. Corneas were monitored by slitlamp, histology, and microbial culture after 24 hr. RESULTS: No CLPU-like lesions were detected under the following conditions: corneal scratch plus lens wear with no bacteria; corneal scratch plus S. epidermidis colonized lenses; corneal scratch without lens wear and with S. aureus applied to eyes; no scratch plus S. aureus colonized lenses; and corneal scratch plus contact lenses colonized by dead S. aureus. CLPU-like lesions were found only when the corneas were scratched and contact lenses colonized by viable S. aureus were applied to the eye. The histology of the lesions demonstrated a frank epithelial break with underlying stromal infiltration. Only low numbers of bacteria could be cultivated from the corneas with CLPU-like lesions. The CLPU-like lesions shared many similarities with CLPU in humans. CONCLUSION: CLPU-like lesions were only produced by S. aureus and not S. epidermidis in the presence of an epithelial abrasion and contact lens. Thus, we hypothesize that to reduce the incidence of CLPU, contact lenses designed to reduce corneal interaction and repel microbial colonization should be produced. PMID- 12772735 TI - Contact lens infections: can they ever be eradicated? AB - Since their introduction, the most significant complication of wearing soft contact lenses has been the development of vision-threatening microbial keratitis. In lens-wearing corneas, microbial infection is thought to develop in the absence of overt injury, leading to the hypothesis that microbe interactions with the corneal epithelium are critical to the pathogenesis of this disease. Thus, we have focused our research efforts on understanding microbial virulence mechanisms aimed at corneal epithelial cells and the innate defenses that normally protect them using the gram-negative bacterium Pseudomonas aeruginosa as a model organism. This report summarizes those results and explores their relevance to understanding contact lens-related infections. PMID- 12772733 TI - Differential diagnosis of microbial keratitis and contact lens-induced peripheral ulcer. AB - Differentiating a contact lens-induced peripheral ulcer (CLPU) from early stage microbial keratitis (MK) is primarily based on clinical judgment rather than on microbiologic or histopathologic investigations. For this reason, tests do not provide valuable information at the early stages in the clinical course of MK. Whereas in gross terms, the clinical picture of MK is more acute and severe than CLPU, clinical features of the two can overlap, sometimes resulting in errors of judgment and mismanagement. This article provides clinical clues that help distinguish the two conditions. In addition, a scoring system has been devised for MK and CLPU. Microbial keratitis (MK) is a dreaded complication for contact lens wearers. Although the risk is small, the large population of contact lens (CL) users have made CL wear a major predisposing factor for corneal infection. The reported incidence of MK among CL wearers may be inaccurate because it can be easily confused with its sterile counter part, contact lens-induced peripheral ulcer (CLPU). An accurate initial clinical impression is critical in avoiding mismanagement of these conditions. Clinical differentiation between infected and sterile corneal infiltrates in CL wearers has been addressed in the literature. We suggest a scheme for distinguishing early stage MK from CLPU. PMID- 12772736 TI - The role of hypoxia in the limbal vascular response to soft contact lens wear. AB - PURPOSE: Increases in limbal vascular blood flow frequently occur as an accompaniment to conventional soft contact lens wear. This study attempted to discover whether these changes can be induced by the direct influence of oxygen alone in the absence of contact lenses. METHODS: Nine people were subjected to monocular anoxia by bathing one randomly chosen eye with 100% nitrogen using gas tight goggles. Their other eyes served as controls and were exposed to normal air throughout the study. Vascular changes were quantified by image capture and subsequent analysis, with the change in the proportion of the image corresponding to blood vessels (per area) being used as the measure of hyperemia. RESULTS: Nitrogen-exposed eyes showed significantly greater hyperemia (mean perarea change +/- 95% confidence interval, 0.023 +/- 0.016 compared with control eyes, -0.013 +/- 0.013; P=0.004, paired t test). CONCLUSION: Reduced oxygen concentration at the ocular surface induces more blood flow in limbal vessels. This result provides further evidence that similar changes occurring during soft contact lens wear are caused by lens-induced hypoxia. PMID- 12772737 TI - Lysozyme and lipid deposition on silicone hydrogel contact lens materials. AB - PURPOSE: We sought to determine whether there were differences in lysozyme (quantity and conformation) and lipid deposition on in vivo worn conventional (etafilcon) and silicone hydrogel (balafilcon and lotrafilcon) contact lenses. METHODS: After extraction, lysozyme concentration in each extract was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. Lysozyme activity was determined by the rate of lysis of Micrococcis lysodeikticus cells. Lipid deposition was determined by high performance liquid chromatography. RESULTS: Lysozyme deposition on etafilcon lenses was significantly greater than that measured on silicone hydrogel (SH) lenses (985 microg per lens versus 10 and 3 microg per lens for balafilcon and lotrafilcon materials, respectively; P<0.001). The degree to which lysozyme was denatured was influenced by the lens material, with the lowest degree of denaturation (22%) seen on the conventional lens material, as compared with 50% for balafilcon and 80% for lotrafilcon (P<0.001). Lipid deposition was greatest on the SH materials, with up to 600 microg per lens of certain lipid classes being deposited on balafilcon, as compared with 20 microg per lens on etafilcon (P<0.001). CONCLUSION: The quantity and conformation of lysozyme and the quantity of lipid deposited on hydrogel contact lenses is significantly influenced by the composition of the lens material. SH contact lens materials deposit low levels of lysozyme and high levels of lipid deposition compared with ionic contact lens materials. Although SH materials deposit only small amounts of lysozyme, the degree of lysozyme denaturation that occurs is higher relative to that seen on ionic lens materials. PMID- 12772738 TI - Tear turnover and immune and inflammatory processes in the open-eye and closed eye environments: relationship to extended wear contact lens use. PMID- 12772739 TI - Contact lens-related microbial keratitis: what can epidemiologic studies tell us? AB - Since the publication of large-scale population based studies to estimate the risks of contact lens-related microbial keratitis, the lens types and modes of wear have changed considerably, with the introduction of silicone hydrogel contact lenses for continuous wear of up to 30 nights and daily disposable contact lenses. It is time to consider epidemiologic studies to estimate the absolute and relative risk of microbial keratitis associated with these new exposures. This article summarizes current knowledge of the estimates and determinants of risk associated with re-useable hydrogel lenses and discusses study design considerations for future studies. The absolute risk of microbial keratitis to the wearer has been remarkably stable since 1989. Risk factors consistently associated with increased risk have included overnight wear, the duration of continuous overnight wear, lower socioeconomic class, smoking, and lens hygiene practice, specifically in daily wear lenses. Men seem to be at a slightly higher risk compared to women. Factors such as the lens age, duration of wear or wear history, and time since the last aftercare visit do not seem to modulate risk. Outstanding questions include whether the risk has changed over time, whether the risk is modified with daily disposable or silicone hydrogel wear, whether the risk is different for 6 or 30 nights of continuous wear, or whether the severity of the disease or spectrum of causative organisms has been modified with new lens types. Different study designs have been used to derive risk data in contact lens wear, including cohort, surveillance, and case-control designs. An appreciation of the advantages and limitations of each of these approaches will assist in the planning of future studies to address these outstanding questions. PMID- 12772740 TI - Tear exchange and oxygen reservoir effects in silicone hydrogel systems. AB - PURPOSE: The effects of oxygen reservoir and tear exchange are known for conventional hydrogel contact lenses. This study attempted to (1) confirm their presence in lenses of a silicone hydrogel (SH) material and (2) evaluate their individual and combined contributions to hypoxic relief of the cornea. METHODS: Corneal oxygen uptake rates were measured polarographically for 10 OD corneas immediately after 300-sec periods (equivalent to 60+ blink cycles) of: (1) nonblink wear of a 0 Dk/L polymethylmethacrylate (PMMA) cap lens; (2) nonblink wear of that cap lens with a SH lens inserted between it and the cornea; (3) regular blinked wear of that same lens combination; (4) nonblinked wear of the SH lens alone; and (5) normal open-eye, non-lens wear. The rates for each eye had a ratio with its own baseline (no lens) rate, and the total sample mean was calculated for each condition from those individual eye means. RESULTS: (1) Under nonblink conditions, a 28% reduction in corneal oxygen demand was observed when a SH lens was inserted under the PMMA cap lens versus without. (2) When regular blinking was added to that lens combination, corneal oxygen demand decreased another 8%, for a total of 36%. The combination of SH insertion and blinking did achieve a statistically significant difference (P<0.05 by the Dunnett test) from the nonblink, 0 Dk/L, maximum deprivation condition. CONCLUSION: The statistically significant hypoxia reduction observed with the SH lens insertion and blinking indicates the additive presence of two factors: (1) a lens reservoir effect caused by the SH lens and (2) a bulk-flow tear exchange effect caused by blinking. Their respective contributions to the reduction of corneal oxygen demand over the period studied were found to be in the ratio of 3.4:1. PMID- 12772742 TI - What is dry eye and what does it mean to the contact lens wearer? AB - PURPOSE: To define dry eye disease and describe the evolution of our understanding of the clinical expression and pathophysiology of dry eye. To relate tear function to contact lens wear and review the problems dry eye causes with contact lens tolerance and performance. METHODS: Review of literature. RESULTS: The rudimentary definition of dry eye as a lack of tears is expanded through an increased understanding of the clinical presentations of dry eye. The classification of dry eye is historically reviewed. The pathophysiology is related to the evolution of therapy for dry eye disorders. The function of the tear film is related to contact lens wear and the problems induced by dry eye correlated with contact lens intolerance. CONCLUSION: The tear film is critical to successful contact lens wear. Disturbances of the quantity or quality of the tear film, whether because of aqueous deficiency or evaporative tear problems, results in intolerance of contact lens wear and damage to the ocular surface. PMID- 12772741 TI - Eyelid disorders: the meibomian gland, blepharitis, and contact lenses. AB - PURPOSE: To present our results to date on the evaluation of the pathophysiology of chronic blepharitis and its relationship to the tear lipid layer, and to propose a structure to that layer. METHODS: Compilation of work to date. RESULTS: A classification system of chronic blepharitis has been developed that has use in diagnosis, planning therapy, and assessment of mechanisms. Abnormalities in meibum have been identified that provide insight into disease mechanisms and have led to the development of a proposed structure to the tear lipid layer. A defect in polar lipids explaining the frequently associated evaporative dry eye has been identified. Mechanisms accounting for the therapeutic benefit of minocycline have been elucidated. CONCLUSION: Chronic blepharitis has associated meibum abnormalities that explain defects in the tear lipid layer that may result in a frankly unstable tear film and that explain the frequently associated evaporative dry eye. PMID- 12772743 TI - Dryness and discomfort with silicone hydrogel contact lenses. AB - PURPOSE: The purpose of this study was to determine whether symptoms of dryness and discomfort are experienced differently with silicone hydrogel lenses compared to conventional hydrogels, in symptomatic and asymptomatic subjects. METHODS: Thirty-nine symptomatic and asymptomatic subjects wore four types of lenses: Focus NIGHT & DAY (CIBA Vision), Focus DAILIES (CIBA Vision), ACUVUE 2 (Johnson & Johnson Visioncare) and Proclear Compatibles (CooperVision) contralaterally for 7 hours and rated comfort and dryness on a zero-to-100 point visual analog scale at 0, 1, 3, 5 and 7 hours. RESULTS: In both groups, no lens differences were found for comfort and dryness, but the comfort and dryness ratings of the symptomatic group decreased significantly (became worse) over the 7-hour period. CONCLUSIONS: These results demonstrate that dryness and comfort is the same over time with silicone hydrogel as with the other three lenses. PMID- 12772744 TI - Interactions of Pseudomonas aeruginosa and Staphylococcus epidermidis in adhesion to a hydrogel. AB - PURPOSE: Bacterial strains with known capability to bind to hydrogel lenses were examined for their relative capacities to attach irreversibly to a hydrogel contact lens in pure and mixed cultures. METHODS AND RESULTS: Time course studies of adhesion through adenosine triphosphate (ATP) or radiolabel analyses indicated that primary adhesion by Pseudomonas aeruginosa to a 37% water-content hydrogel lens was more dense and rapid than that of Staphylococcus epidermidis. Primary adhesion of P. aeruginosa was similar after 5 min and 2 hr, whereas S. epidermidis showed an incremental increase in adhesion over the first 30 min. Sequential challenge of lenses with P. aeruginosa followed by S. epidermidis gave levels of primary adhesion for each species similar to those obtained with single pure cultures. When lenses were challenged with S. epidermidis first, primary adhesion of P. aeruginosa was significantly diminished. Exposures of multiple sets of lenses (n = 4) to a standard inoculum of P. aeruginosa resulted in levels of primary adhesion per lens after 1 hr equivalent to that found for a single lens in 5 min or in 2 hr. CONCLUSION: These data further support that normal biota may be protective for the eye. The data also suggest that under conditions of the primary adhesion-screening test, a limited number of cells of GSU no. 3 (selected phenotype) are involved in adhesion to the hydrogel. PMID- 12772745 TI - Can postlens tear thickness be measured using three-dimensional in vivo confocal microscopy? AB - PURPOSE: Confocal microscopy through focusing (CMTF) of the cornea produces a three-dimensional display of corneal structure and intensity profiles that allow objective measurements of corneal sublayer thickness. This study investigated the feasibility of using in vivo CMTF to measure postlens tear thickness (PLTT). METHODS: Two rabbits and one human were evaluated in this study. Both rigid gas permeable (RGP) and silicone hydrogel contact lenses were used. After contact lens insertion, CMTF scans were performed by rapidly focusing from the tip of the objective, through the contact lens, PLTT, and full thickness of the cornea, and into the anterior chamber of the eye. PLTT was calculated by measuring the distance between the posterior surface of the lens and the anterior surface of the epithelium. RESULTS: With a steep RGP lenses, the position of the posterior lens surface and the superficial epithelial surface was easily visualized using three-dimensional CMTF, and the PLTT could be measured. However, when a properly fit RGP or silicone hydrogel contact lens was used, the strong signal from the posterior surface of the lens washed out the signal from the front surface of the cornea in most cases, and the PLTT could not be directly assessed. We also found that the PLTT was sensitive to applanation of the confocal objective lens. CONCLUSION: The data suggest that the tear film under a properly fit contact lens is less than the 9-microm axial resolution of our confocal microscope. Furthermore, the potential for undetected eye movement and accidental touching of the contact lens raises questions regarding the overall reliability of the measurements. PMID- 12772746 TI - The case for rigid lenses. AB - The case is made for rigid lenses in the face of increasing market share for soft lenses. Safety, visual function, long-term comfort and durability are key positive features for these devices. PMID- 12772747 TI - The case against rigid contact lenses. AB - Rigid lens fitting has been an integral part of contact lens practice for decades. However, rigid lens fitting has been constantly declining since soft lenses were introduced in the 1970s to whereby, world wide, rigid lenses constitute less than 10% of new contact lens fits. It seems that many practitioners correctly or incorrectly believe that rigid lenses only need to be prescribed in specialized or extenuating circumstances. This discussion article examines the international decline in rigid lens fitting and seeks to explain the cause of this trend. I conclude that rigid lenses will be virtually obsolete by the year 2010. PMID- 12772748 TI - Comfort response to rigid and soft hyper-transmissible contact lenses used for continuous wear. AB - PURPOSE: To assess subjective comfort during the first three months of continuous wear of hyper-transmissible soft and rigid contact lenses. METHODS: One hundred subjects (50 experienced wearers and 50 neophytes) were fitted with either the Focus Night & Day lens (CIBA Vision, Duluth, GA) or the Z alpha lens (Menicon, Nagoya, Japan). Comfort was monitored at regular intervals using a vertical analog comfort scale. RESULTS: Good comfort scores (generally indicating very comfortable lenses) were recorded for the two soft lens groups (experienced and neophytes) and the experienced rigid lens group; these scores remained approximately constant throughout the study. Comfort reported by neophyte rigid lens wearers was initially recorded as being between slightly uncomfortable and comfortable; however, after 1 week of daily wear and 1 night of overnight wear, the mean comfort score of this group reached a level that was similar to that recorded by subjects in the other three study groups. CONCLUSION: The comfort data presented in this study can be used by clinicians to anticipate the subjective response of both new and existing lens wearers to new-generation hyper transmissible soft and rigid contact lenses. PMID- 12772749 TI - Microbial keratitis and vision loss with contact lenses. AB - PURPOSE: Microbial keratitis is the only sight-threatening adverse event that occurs with contact lens wear. This article gives a preliminary estimation of the incidence of microbial keratitis and vision loss with continuous-wear contact lenses made from highly oxygen permeable (Dk) materials. METHODS: The most up-to date data available on microbial keratitis and vision loss are collected from a range of sources including industry, private practice, and publications and is reviewed. RESULTS: There have been 16 cases of microbial keratitis with high-Dk silicone hydrogel lenses. Of the 13 where data are available, none have lost two or more lines of best-corrected visual acuity (BCVA). CONCLUSION: First approximation indicates that the incidence of microbial keratitis with high-Dk silicone hydrogel lenses may be lower than the incidence with low-Dk soft lenses during extended wear. The rate of loss of more than two lines of BCVA is low in patients that develop microbial keratitis with low- and high-Dk soft lenses. PMID- 12772750 TI - Loss of vision after laser in situ keratomileusis. AB - PURPOSE: To research the incidence of visual loss after laser in situ keratomileusis (LASIK). METHODS: We reviewed an extensive review of the literature published by The Ophthalmic Technology Assessment Committee of the American Academy of Ophthalmology regarding the safety and efficacy of LASIK. We also reviewed other recent literature. RESULTS: For low to moderate myopia and astigmatism, LASIK is efficient, safe, and predictable with few eyes losing two or more lines of best-corrected visual acuity (BCVA). For moderate to high myopia, the results are more variable and more eyes lost two or more lines BCVA. In both cases, this BCVA occurred in studies published on or before 1999. CONCLUSION: Serious adverse complications leading to significant visual loss are probably rare. Continued development of microkeratomes and laser technology are continuing to reduce the rare complications that do occur and improve functional outcomes and patient satisfaction. PMID- 12772751 TI - Are we nearsighted when it comes to myopia treatment? AB - PURPOSE: The purpose of this article is to highlight the current research on myopia treatment and to speculate about the future of myopia treatment. METHODS: A review of the literature on myopia treatment in humans was performed using Medline, CRISP, and a variety of web pages. A summary of the research is presented in the article. RESULTS: Most of myopia research can be categorized into three areas: control, treatment, and prevention. Most of independent myopia research currently conducted is in the area of control. Great advances in the myopia treatment are constantly being practiced, but few independent investigations are conducted. Methods of predicting the future onset of myopia are being investigated to some day allow us to apply preventive agents before myopia occurs. CONCLUSION: Two randomized clinical trials are scheduled for completion within the next year, and they should offer answers about mechanisms of myopia control. Treatments for myopia are moving beyond questions of safety and efficacy and on to reductions of complications and refinement of vision to produce super vision. At least one eye drop, pirenzepine, is being investigated for myopia control and may show promise for prevention of myopia in the future. PMID- 12772752 TI - Why daily wear is still better than extended wear. AB - Among the many problems complicating contact lens wear, microbial keratitis is of most concern because of its potential for substantial morbidity. Three decades of basic and clinical research suggest that risk factors include poor care compliance and extended wear through one or multiple sleep cycles. Many believe the latter problem is caused in part or in whole to contact lens-induced hypoxia. New contact lenses, both rigid and soft, have been developed that allow oxygen delivery equivalent to the noncontact lens state, under open-eye conditions and close to the same even for closed-eye conditions. But will such lenses reduce the risk for microbial keratitis? The authors argue that until such a conclusion is reached through clinical trials, the question remains in doubt. PMID- 12772753 TI - The case for continuous wear. PMID- 12772754 TI - Menicon Z 30-day continuous wear lenses: a clinical comparison to Acuvue 7-day extended wear lenses. AB - PURPOSE: To establish equivalent safety and efficacy of the clinical performance of a novel hyper-O2 transmissible rigid gas-permeable (RGP) lens (Menicon Z, tisilfocon A) when worn continuously for 30 days (29 nights) when compared with a conventional control hydrogel lens (Acuvue; etafilcon A) worn for 7 days (6 nights). METHODS: This study was a prospective, open-label, 24-center, concurrent, cohort-controlled clinical trial. This abbreviated report will focus on the length of wear achieved, slitlamp findings, and adverse event rates. RESULTS: Two-thirds (66.5%) of patients wearing the RGP lens achieved greater than or equal to 22 nights of continuous wear; slitlamp observations revealed two categories of findings: those associated with lens modality and those with lens wearing time. Adverse events were mostly associated with foreign body abrasions for RGP lenses, whereas the hydrogel lens showed a higher number of bacterial infections. CONCLUSION: The results of this study revealed some interesting differences in the classification and severity of findings. The results demonstrate that the continuous wear of Menicon Z (tisilfocon A) RGP lenses for up to 30 nights is a safe and equivalent alternative to 7-day (6 nights) hydrogel wear lenses. PMID- 12772755 TI - Risk factors for corneal infiltrative events with 30-night continuous wear of silicone hydrogel lenses. AB - PURPOSE: This analysis reports on risk factors for corneal infiltrative events from a 1-year, randomized clinical trial of lotrafilcon A lenses. METHODS: Six hundred fifty-eight subjects randomized to lotrafilcon A lenses were observed in a 1-year clinical trial in the United States. Risk factors gathered at baseline were analyzed to determine their association with corneal infiltrative events. RESULTS: Factors examined but not found to be associated with events were male sex, history of extended or daily lens wear, refractive error, neovascularization, or assessment of lens fit. Significant factors included subject age 18 to 29 years, smoking, smoking and young age combined, history of corneal scar, contact lens acute red eye (CLARE), and corneal infiltrates. CONCLUSION: Risk factors for corneal infiltrative events with silicone hydrogel lenses worn for 30 nights of continuous wear have been identified. Wearers with a history of inflammatory events such as contact lens peripheral ulcer or CLARE should consider 30-night continuous wear with these lenses. Careful counseling of wearers who smoke is advised. PMID- 12772756 TI - Assessing the safety of the new 30-night contact lenses. PMID- 12772757 TI - The U. S. Food and Drug Administration's role in contact lens development and safety. AB - PURPOSE: To provide information concerning the U. S. Food and Drug Administration's (FDA) role as a regulatory agency in the development and safe use of contact lenses. METHODS: The FDA's organizational structure and regulatory classification of medical devices are explained. Specific regulatory authorities are described and examples provided with respect to the FDA's role in the regulation of the new hyper-oxygen permeable continuous wear contact lenses. RESULTS: The application of medical device regulations addresses the rights and safety of subjects in preapproval clinical studies and provides a basis to generate valid scientific evidence to demonstrate the safety and effectiveness of a medical device. Once on the market, there are postmarket requirements with which a manufacturer must comply. CONCLUSION: For continuous wear 30-day lenses, the FDA is using additional regulatory authorities new to the contact lens area to assure that product safety is maintained and consumer information is improved. These include postapproval studies, more informative labeling, and advertising restrictions. Whereas it is the FDA's mission to promote and protect the health of the public by ensuring the safety and effectiveness of medical devices, the agency works in partnership with other stakeholders to maintain the safe use of medical devices. PMID- 12772758 TI - Design principles and limitations of wave-front guided contact lenses. AB - The concept of the wave-front guided design of contact lenses is presented from three vantage points: ray optics, wave front aberrations, and optical path-length errors. We argue that the goal of contact lenses is to make all of the optical paths from a distant object to the retina equal in length, regardless of where the path intersects the plane of the eye's pupil. The aberration map of an eye is a prescription for such a lens. Unfortunately, variability of measured aberration maps is a fundamental limit to our knowledge of the true aberration structure of an eye. Variability arises because the eye is a biologic system that changes over time for normal, physiologic reasons. Furthermore, uncertainty in our measurement of the aberration map because of such variable factors, such as alignment of the aberrometer to the eye by the clinician or small fixation errors committed by the patient, will make it difficult to achieve a full measure of success with aberration-correcting contact lenses. The clinical implication of these findings is that multiple measurements of the aberration map should be collected using a protocol that includes realignment of the instrument and then averaging the aberration maps to reduce the level of uncertainty associated with any single measurement. PMID- 12772759 TI - Toric contact lens designs in hyper-oxygen materials. AB - A high proportion of eyes show significant astigmatism, and therefore toric silicone hydrogel lenses are required to satisfy the demand for continuous wear. Two concerns arise from the inevitable, increased thickness associated with toric designs: reduced oxygen transmissibility and increased rigidity leading to possible mechanical trauma. An improved understanding of the mechanisms governing toric soft-lens orientation and fit will enable the development of thinner designs. Gravity has little effect on prism-ballasted soft lenses within approximately 30 degrees of the base-down position. Some correlations have been noted between lid position and orientation. Measurements from video recordings of toric soft lenses during reorientation indicate that lid-induced rotation takes place during rather than between blinks. However, high-speed video recordings indicate that the influence of the lower lid can override that of the upper lid depending on its position, tightness, and amount of lateral movement. This knowledge gives some pointers toward optimizing toric soft lens designs in hyper oxygen materials. PMID- 12772761 TI - New bifocal designs in hyper-oxygen materials. AB - PURPOSE: To address the question, "How can we increase the bifocal contact lens market?" METHODS: The design, lens parameters, and the results of a clinical study with this new bifocal lens are discussed. The contact lens market and current bifocal contact lens designs are reviewed and factors that should be considered in future lens designs are recommended. RESULTS: The only hyper-oxygen bifocal available is the Menifocal Z (Menicon, Nagoya, Japan). CONCLUSION: Simultaneous vision, the optical design used in most soft lens bifocals, reduces visual performance. Industry and vision scientists need to develop new bifocal lens designs to help the bifocal contact lens market grow. PMID- 12772760 TI - Can UV radiation-blocking soft contact lenses attenuate UV radiation to safe levels during summer months in the southern United States? AB - PURPOSE: Peak solar UV radiation (UVR) intensities are typically experienced in summer months. People living in the southern states of the United States, where the UVR frequently exceeds the recommended minimum erythema dose (MED), are at particular risk, especially outdoor workers. The present study analyzed summertime MED readings in Houston, TX, to assess the frequency of intensities regarded as unhealthy. The study also sought to assess whether UV-blocking hydrogel contact lenses provide ocular protection from these high doses. METHODS: Readings, taken at midday using a UVR biometer, were analyzed to assess the potential UVR risk. The spectral response of the meter, modified by the spectral transmission curves of the contact lenses, allowed us to mathematically assess the ocular protection provided. In addition, ambient UVR measurements were taken at midday, using a portable UVR radiometer. The detector was adapted so that a standard diameter hydrogel contact lens could be placed over it to quantify the UV-blocking capabilities of the lens. RESULTS: The MED readings showed that the recommended safety standards were exceeded approximately at local midday 90% of the time. Model calculations and empirical data demonstrated that contact lenses attenuated the MED readings by up to 90%, bringing them well within the recommended Environmental Protection Agency safety standards. CONCLUSION: The efficacy of the model used in this study was verified through direct comparison of the modeled and measured data. UV-blocking hydrogel soft contact lenses reduce the MED to the human eye and therefore limit the lifetime ocular dose. These lenses are highly recommended to prevent the development of UVR-related ocular pathologic conditions. PMID- 12772762 TI - Aberration correction with soft contact lens: is the postlens tear film important? AB - PURPOSE: A number of factors may influence the performance of soft contact lenses designed to correct the aberrations of the eye. Although the effect of many of these factors has been investigated, the effect of postlens tear film has not. We estimated the optical effect of the postlens tear film by using simplified models of postlens tear thickness profiles. METHODS: Three models were analyzed, including (1) a worst-case scenario before lens settling, (2) representative components of thickness profile based on Zernike-like forms, and (3) a series of undulations (ripples) across the postlens tear film. A range of thicknesses of the tear film was studied. Ray-tracing analyses were conducted on all models to calculate the modulation transfer function response between 0 and 100 cycles per degree and Strehl ratio. RESULTS: The results show that in only a few scenarios does the reduction in optical performance because of the postlens tear film approach those introduced by conventional spherical surface soft contact lenses. CONCLUSION: The postlens tear film can introduce discernable amounts of optical degradation in the lens-wearing eye. However, the amount of this effect is likely to be small compared with other sources, and benefits afforded by the use of aberration-corrected soft contact lenses should not be severely affected. PMID- 12772763 TI - Noninflammatory silicone hydrogel contact lens complications. AB - High Dk silicone hydrogel lenses have overcome many of the hypoxic problems associated with traditional extended wear, and the popularity of continuous wear with these lens types is increasing. Despite the elimination of hypoxia, several noninflammatory clinical complications have been reported to occur as a result of mechanical disturbances or trauma when wearing silicone hydrogel lenses. This article reviews the origin and presentation of mucin balls, superior epithelial arcuate lesions, contact lens papillary conjunctivitis, and corneal erosions in silicone hydrogel lens wearers. A number of management strategies and approaches to minimize the occurrence of these clinical complications are also discussed. PMID- 12772764 TI - Righteousness and reason. PMID- 12772765 TI - Transnational licensure: view from Texas. AB - Texas has a diverse population with many underserved communities and a dentist-to population ratio only 80% of the national average. Currently dentists trained in other countries can be licensed after completing four years of dental school and passing either the Western or Central Regional Testing Association's examination or by completing a graduate program in an ADA-accredited specialty. Although there have been careful and lengthy analyses of comparability of training across countries, a fundamental issue is that standards for education are developed by a national voluntary organization (Commission on Dental Accreditation) and standards for licensure are determined by individual states in the United States. Elsewhere in the world, the federal government performs these roles. PMID- 12772766 TI - Portability of licensure in Canada based on accreditation and certification. AB - Although registration and licensure of dentists in Canada is the purview of the ten provincial licensing authorities in Canada, establishment of standards and certification of candidates has for fifty years been the responsibility of the National Dental Examining Board. The NDEB administers a validated set of written and objective standardized clinical examinations to educationally qualified candidates. Such candidates include those who have graduated from accredited Canadian and United States schools and graduates of non-accredited schools who have satisfactorily finished a qualifying program. The NDEB cooperates closely with U.S. educational and accreditation programs and the American Dental Association. PMID- 12772768 TI - Transnational licensure: foreign dentists in America reclaim their profession through the Program for Advanced Standing Students (PASS). AB - The Program for Advanced Standing Students at the University of Pennsylvania School of Dental Medicine accepts highly qualified individuals who have earned a dental degree in a foreign country. They earn an American dental degree in two years, following a program that is nearly identical to the final years of the traditional DMD program. PASS students have been highly successful in their educational program and they contribute in diverse ways to the dental profession upon graduation. PMID- 12772767 TI - Towards higher standards in dental and stomatological education in Europe. AB - In the creation of the European Union, attention was given to portability of licensure for professionals. Considerable differences exist among countries in culture, economic conditions, and educational resources and practices. In dentistry, these differences in professional training have been addressed through a peer consultative process rather than through political and legal means. The process of visits to dental schools throughout Europe and the organizational structure (DentEd) used to conduct the visits and summarize findings are described. PMID- 12772769 TI - A retrospective of America's second National Dental Association. AB - This article describes one hundred and fifty years of history concerning organized dentistry among African Americans. African Americans were welcomed in the National Negro Medical Association of Physicians, Dentists and Pharmacists (NMA) as early as 1895 and later created a dental professional group, the Interstate Dental Association, in the mid-Atlantic states. The "second" National Dental Association, a national-wide African American association was created in 1932. African Americans earned full membership in the American Dental Association in 1964. This article traces these developments against the background of changing American society and concludes by mentioning some of the recent accomplishments and collaborative relationships of the current NDA. PMID- 12772770 TI - My way or the highway: do dental patients really have autonomy? AB - Ethical dilemmas arise when it is not possible to simultaneously optimize conflicting values, each of which independently is worthy. This paper analyzes three cases where patients' autonomy is in conflict with dentists' professional judgments about their own practice patterns and what is in the best interests of patients' oral health. The hierarchy of values proposed by Ozar and Sokol is a valuable aid in addressing such dilemmas, but the dentist must still engage in a detailed analysis of the situation. PMID- 12772771 TI - The mumpsimus. AB - It is an unfortunate side of human nature, on occasion, to stubbornly refuse to change our minds or listen to evidence that challenges our beliefs. In the extreme form, such individuals are called mumpsimuses. This column describes what research has been able to tell us about why individuals continue to escalate their commitments to failing causes and why beliefs are so resistant to change, even in the light of reason. In fact, it is usually best to leave a sleeping mumpsimus lie. PMID- 12772772 TI - Participation of corticosteroids and effects of indomethacin on the acute inflammatory response of rats fed n-6 or n-3 polyunsaturated fatty acid-rich diets. AB - We have previously shown that both n-3 (fish oil) and n-6 (soybean oil) PUFA-rich diets reduce carrageenan-induced paw edema in rats. The present study evaluated the role of corticosteroids, and the effect of indomethacin on this response. Basal (pre-carrageenan) levels of corticosterone were elevated in both lipid diets compared to the chow diet. During inflammation, corticosterone levels increased to a similar extent in the chow and lipid diets. With 2.0 mg/kg indomethacin, edema was reduced in the chow diet and the n-3 diet, while it was not changed in the n-6 diet. In contrast, the 16.6 mg/kg dose of indomethacin induced a mild increase in edema in the chow diet but a pronounced edema increase in the lipid diets. The increase in corticosterone levels induced by carrageenan was either reduced (chow) or completely abolished (lipids) by the treatment with the higher dose of indomethacin, compared to both the control (untreated) group, and the lower dose of indomethacin. These data indicate that both acute inflammation and the response to an antiinflammatory drug were attenuated by n-3 or n-6 PUFA-rich diets. They also showed that indomethacin can have anti- or proinflammatory properties reflecting the extent of the corticosterone inhibition by indomethacin. PMID- 12772773 TI - Glucocorticoids modulate TGF-beta production by human fetal lung fibroblasts. AB - TGF-beta1 is thought to play a central role in pulmonary fibrosis inducing fibroblast differentiation and extracellular matrix synthesis. In human lung fibroblasts, it is still unclear how various TGF-beta isoforms affect TGF-beta production and whether glucocorticoids, commonly used agents to treat fibrotic lung disease, modulate these processes. To this end, human fetal lung fibroblasts (HFLF) were cultured with various concentrations of glucocorticoids (budesonide, dexamethasone or hydrocortisone) with and without TGF-beta1, -beta2, or -beta3. Post-culture media were collected for ELISA assays of TGF-beta1, -beta2, and beta3. TGF-beta mRNA was assessed by real time RT-PCR. Smad 2, 3, and 4 and AP-1 complex (c-fos and c-Jun) cellular localization were evaluated by immunostaining. TFG-beta2 and -beta3 stimulated TGF-beta1 production significantly (p < 0.01 relative to control). TGF-beta1 stimulated TGF-beta2 production (p < 0.01 relative to control). TGF-beta3 was undetectable. Glucocorticoids significantly inhibited TGF-beta1 and TGF-beta2 production and reduced expression of the up regulated TGF-beta1 and TGF-beta2 mRNA induced by exogenous TGF-beta1, -beta2, or -beta3 (p < 0.01 for each) but had no effect on Smads. Although c-jun-related nuclear staining was not intensified in TGF-beta-stimulated cells, it was reduced by glucocorticoids. Thus, TGF-beta isoforms may stimulate production of various TGF-beta isoforms in the lung. Glucocorticoids then may block TGF-beta production by modulating mRNA levels and c-Jun. PMID- 12772774 TI - Selective COX-2 inhibition is associated with decreased mucosal damage induced by acid and pepsin in rabbit esophagitis. AB - OBJECTIVE: To evaluate the effects of COX-1 and/or COX-2 inhibition in a model of chronic esophagitis in rabbits. METHODS: Both high- and low-grade esophagitis were induced in rabbits by the perfusion of acidified pepsin. Rabbits were treated with either a selective COX-2 inhibitor (DFU[3-(3-Fluorophenyl)-4-(4 Methanesulfonyl)-5,5-Dimethyl-5H-Furan-2-One];30 mg/Kg/day), a nonspecific COX inhibitor (indomethacin; 2 mg/Kg/day), or a COX-1 preferential inhibitor (piroxicam; 2 mg/Kg/12 h). RESULTS: Prostaglandins are derived from COX-1 activity in the normal esophagus. Both low- and high-grade esophagitis are associated with a progressive increase of COX activity, which is partially dependent on the COX-2 isoform. DFU reduced muscosal damage in both models of esophagitis. However, indomethacin did not affect significantly mucosal damage, and piroxicam increased damage in low-grade esophagitis. CONCLUSIONS: COX-1 activity is constitutive in the rabbit esophageal mucosa, but both COX-2 and COX 1 activity are increased under the impact of acidified pepsin. Treatment with the COX-2 inhibitor DFU is associated with improvement of mucosal damage, which may have therapeutic implications. PMID- 12772775 TI - Characterization of the in vitro anti-inflammatory activity of AL-5898 and related benzopyranyl esters and amides. AB - Selected ester- (AL-5898 and AL-8417) and amide-linked benzopyran analogues (AL 7538 and AL-12615) were evaluated in vitro for their ability to inhibit key enzymes/processes of the inflammatory response. AL-7538 and AL-12615 exhibited weak intrinsic cyclooxygenase inhibitory activity (IC50 = 13 microM, 37 microM). In contrast, 5-HETE and LTB4 synthesis in A(23187)-stimulated neutrophils was effectively inhibited by both ester and amide analogs (IC50 = 2-3 microM). While there was some indication for differing sensitivities among benzopyran esters and amides in the suppression of cytokine synthesis in stimulated U-937 cells, there appeared to be no great discrimination when assessing their effect on U-937 cell adhesion to IL-1beta activated HMVEC-L cells. Inhibition of cell adhesion was concentration-dependent, with IC50 values ranging between 18 microM and 30 microM for AL-5898. Concentration-dependent inhibition of inflammatory cytokine production (i.e., IL-1beta, TNF-alpha, GM-CSF and IL-6) was also apparent in LPS stimulated, cultured PBMC as well as in PMA/A(23187) activated U-937 cells monitoring the synthesis of IL-1beta, IL-8, TNF-alpha, and MCP-1. Notably, the hydrolysis products of the benzopyranyl ester, AL-5692 and (S)-6-methoxy-alpha methyl-2-naphthaleneacetic acid, were devoid of pharmacological activity when assessed for inhibition of monocyte adhesion or IL-1beta synthesis. Collectively, our data demonstrate the unique in vitro polypharmacology of a novel series of benzopyran analogs that suppress pivotal enzymes and processes in the inflammatory response. PMID- 12772776 TI - Inhibition of the neutrophil NADPH oxidase by adenosine is associated with increased movement of flavocytochrome b between subcellular fractions. AB - Adenosine is a potent inhibitor of reactive oxygen species (ROS) production by the NADPH oxidase in fMLF-stimulated neutrophils. Although much is known about the pharamacology and signal transduction of this effect, it is not known how adenosine affects assembly and localization of the NADPH oxidase components within the neutrophil. We report here that adenosine pretreatment of fMLF stimulated neutrophils results in decreased plasma membrane/secretory granule content of the flavocytochrome b components (p22phox and gp91phox) of the NADPH oxidase, which correlates with inhibition of ROS production. Adenosine treatment did not affect upregulation of secretory and specific granule surface markers, confirming that degranulation was not impaired by adenosine. However, adenosine treatment did result in increased movement of cell-surface flavocytochrome b to heavy granule fractions in fMLF-stimulated neutrophils. These data suggest that adenosine-mediated effects on neutrophil ROS production are due, in part to endocytosis and/or redistribution of flavocytochrome b between various subcellular compartments. PMID- 12772777 TI - Treatment of gastroparesis with electrical stimulation. AB - Electrically stimulating the stomach to treat gastroparesis has been proposed by investigators for decades. With the development of techniques of implantable pacing devices and electrodes and promising preliminary results in chronic pacing studies, gastric electrical stimulation (GES) has received increasing attention recently among researchers and clinicians. A number of studies have been performed to investigate the effects of GES on gastric motility, gastric emptying, and gastrointestinal symptoms in both dogs and humans. Based on the frequency of the electrical stimulus used for chronic treatment of gastroparesis, gastric electrical stimulation can be classified into low-frequency stimulation (LFS) and high-frequency stimulation (HFS). Although some of the results are still controversial, the majority of these studies seem to indicate that LFS is able to normalize gastric dysrhythmias and entrain gastric slow waves and accelerate gastric emptying. On the other hand, HFS has no effect on gastric emptying but is able to significantly reduce symptoms of nausea and vomiting in gastroparetic patients. GES has provided an exciting new advance in the treatment of gastroparesis and management of upper gastrointestinal symptoms. This paper will review the available studies of GES in the treatment of gastroparesis and current status of this field. PMID- 12772778 TI - Constipation and lack of colonic interstitial cells of Cajal. PMID- 12772779 TI - Gene therapy for cancer: current status and prospects. PMID- 12772780 TI - Expression of EGF-receptor related protein (ERRP) decreases in gastric mucosa during aging and carcinogenesis. AB - Aging and gastrointestinal malignancies, including that of the stomach are associated with increased activation of EGF-receptor (EGFR). Although the intracellular events that regulate this process are poorly understood, we hypothesize that loss of ERRP (EGFR-related protein; GenBank accession number AF187818), a recently identified negative regulator of EGFR, that possesses a substantial homology to the ligand binding extracellular domain of EGFR, may contribute to this event. In support of our hypothesis, we have observed that in Fischer-344 rats, whereas aging is associated with increased activation of EGFR in the gastric mucosa, expression of ERRP decreases inthis tissue during this period. The latter is accompanied by a concomitant reduction in the amount of TGF alpha bound to ERRP. In contrast, the amount of TGF-alpha bound to EGFR is found to be higher in the gastric mucosa of aged than in young rats. This is accompanied by a concomitant rise in EGFR levels. In the gastric mucosa, EGFR and ERRP are found to be colocalized. Gastric adenocarcinoma in humans, which has been shown to be associated with increased activation of EGFR, shows a substantial reduction in ERRP expression, when compared with benign tissues. We conclude that increased activation of EGFR in the gastric mucosa during aging and carcinogenesis may partly be due to the loss of ERRP. PMID- 12772781 TI - Detection of mutant p53 in hepatocellular cancer from Turkey and its correlation with clinicopathologic parameters. AB - The samples of hepatocellular carcinoma from Turkey, a country with a high prevalence of hepatitis B virus and hepatitis C virus, but low dietary exposure to aflatoxin B1, were examined in order to detect the frequency of mutant p53 and its association with clinical and pathological data. Fifty-two samples of hepatocellular cancer from the patients who were diagnosed in our clinic were included in this study. The mutant p53 protein was searched for by specific enzyme-linked immunosorbent assay. Of 52 patients with hepatocellular carcinoma, 26 (50%) had the mutant p53. The incidence of p53 mutation in hepatocellular cancer patients with chronic liver disease due to hepatitis B virus infection was significantly higher than in those with chronic liver disease due to alcohol, indicating that not alcohol but hepatitis B virus, in fact induces the mutations in p53 gene. In addition, it has been shown that the p53 mutation was significantly associated with the diameter of tumor nodule and the degree of cellular differentiation in hepatocellular cancer. The p53 mutation rate found in our study is concordant for a geography where hepatitis B virus and hepatitis C virus are common. Hepatitis B virus and possibly hepatitis C virus, but not alcohol, should be responsible, to a degree, for the mutational change in p53 protein in hepatocellular cancer patients with chronic liver disease. The p53 mutation is a late event in hepatocarcinogenesis because it is related with cellular differentiation and tumor diameter. The specific ELISA can be a useful screening test in future studies to select the patients for gene therapy using wild-type p53. PMID- 12772782 TI - Intestinal polyp formation in the Apcmin mouse: effects of levels of dietary calcium and altered vitamin D homeostasis. AB - This study evaluated the effects of various levels of dietary calcium on polyp formation, vitamin D homeostasis, and fecal bile acids in the Apcmin mouse. Female Apcmin mice were randomized to three groups and fed a purified diet with either half or double the level of calcium in control AIN-93G. Serum 25-OH-D and fecal bile acids were measured at weeks 0 and 12 of treatment. Mice were killed for polyp scoring by two observers blinded to treatment after 12 weeks. Results show there was no difference in polyp number or tumor load with dietary calcium in any treatment group. Serum 25-OH-D was reduced and total fecal bile acids were increased in animals that received the high calcium diet. We have previously shown that vitamin D supplementation diminishes polyp load; the lack of effect of an altered calcium diet seen here may be due to a disturbance in vitamin D homeostasis. PMID- 12772783 TI - Systemic combined chemotherapy with low dose of 5-fluorouracil, cisplatin, and interferon-alpha for advanced hepatocellular carcinoma: a pilot study. AB - The purpose of this pilot study was to evaluate the efficacy and adverse events of systemic combined chemotherapy with low dose of 5-fluorouracil (250 mg/m2, 5 days), cisplatin (10 mg/m2, 5 days), and interferon-alpha (2.5 million units, three times weekly) for advanced hepatocellular carcinoma (HCC) underlying liver cirrhosis. Six patients who had advanced HCC with tumor thrombi in the main portal trunk were enrolled in this study. Partial response was achieved in 2, stable disease in 1, and disease progressed in 3. Objective responses were achieved in 2 (33%), however, marked decreases of alpha-fetoprotein protein and protein-induced vitamin K antagonist or absence (PIVKAII) levels were also seen in one patient (stable disease). Four patients showed hematologic or renal toxicity, which were well tolerated and managed. Our systemic combined chemotherapy resulted in favorable response and was well tolerated in those with advanced HCC underlying liver cirrhosis, complicated by leukocytopenia and thromobocytopenia. PMID- 12772784 TI - Effect of N-acetylcysteine on microcirculation of mucosa in rat ileum in a model of intestinal inflammation. AB - Oxygen radicals are formed by the endothelium and blood cells and have specific functions in various organs systems. On the level of the microcirculation, oxygen radicals take part in the regulation of the leukocyte-endothelial interaction. The involvement of oxygen radicals has previously been found in conditions such as sepsis, ischemia-reperfusion, and inflammation. Indomethacin is a clinically applied nonsteroidal antiphlogistic, and in previous studies in the rat, it has been found to induce an inflammatory reaction in the small intestine characterized by edema and reddening of the intestinal epithelium, ulceration, and dysregulation in the intestinal-epithelial barrier function. In the present study, we investigated the effect of N-acetylcysteine on erythrocyte velocity and the arteriolar diameter of the main arteriole in single villi, thus providing insight in the perfusion of the mucosa in indomethacin-induced intestinal inflammation. N-Acetylcysteine is known to inactivate superoxide and its precursors. Therefore, we used N-acetylcysteine to investigate whether superoxide and its precursors participate in the regulation of blood supply to single villi in this animal model. We found that indomethacin induced an increase in villous perfusion that was significantly reduced by N-acetylcysteine, indicating that superoxide and its precursors may participate in the regulation of blood supply to the mucosa in this animal model of intestinal inflammation. PMID- 12772785 TI - Gallbladder ejection fraction and symptom outcome in patients with acalculous biliary-like pain. AB - Patients with acalculous biliary-like pain present a difficult clinical challenge. Our aim was to evaluate the outcome of patients with recurrent biliary like pain without gallstones who underwent testing of gallbladder ejection fraction (GBEF) by cholecystokinin-cholescintigraphy (CCK-CS) in order to determine clinical factors that may predict symptom resolution. We reviewed the records of patients with recurrent acalculous biliary-like pain who underwent CCK CS from January 1995 to December 1999. For comparison, we also studied an age- and sex-matched group of patients who underwent cholecystectomy for symptomatic cholelithiasis. Outcome was obtained by telephone interview, using a scale from 0 to 3 where 0 = no improvement and 3 = clinical remission. Patient demographics, predominant symptom(s), method of management, gallbladder pathology, and response to treatment were recorded. One hundred twenty-nine patients underwent CCK-CS. Of 69 with an abnormal GBEF, 48 (70%) were available for interview. Forty patients underwent cholecystectomy. Twenty-seven patients reported symptom resolution after surgery while 4 nonsurgical patients reported the same (P = NS). Univariate analysis revealed no association between symptom outcome and presence of gastrointestinal symptom(s), severity and duration of abdominal pain, management, or gallbladder pathology. In addition, no GBEF cutoff level predicted symptom outcome. Of the remaining 60 patients with a normal GBEF, 30 (50%) were available for interview. Twenty-eight patients in this group were managed medically and 2 patients underwent cholecystectomy. Eighteen patients managed medically were asymptomatic, as were the 2 who underwent cholecystectomy. There was no difference in symptom outcome between patients who had GBEF >35% vs <35%. In conclusion, in a group of patients with recurrent acalculous biliary-like pain who underwent CCK-CS, we found a high rate of symptom resolution following cholecystectomy; however, this was not statistically different from a smaller cohort who did not undergo surgery. We were unable to determine any variable predictive of symptom resolution. PMID- 12772787 TI - Cytotoxicity of streptozotocin on neuroendocrine cells of the pancreas and the gut. AB - Streptozotocin has been used to induce diabetes mellitus in experimental animals and has been thought to have a selective cytotoxic effect on the beta-cells in the islets of Langerhans. The aim of the present study was to determine whether streptozotocin has any cytotoxic effect on other neuroendocrine cells of the gastrointestinal tract. Eight female Sprague-Dawley rats received intraperitoneal injections of 100 mg/kg streptozotocin in citric acid buffer; the concentration of streptozotocin was adjusted to 25 mg/ml buffer. Seven rats, serving as controls, received an equivalent volume of the vehicle. The rats were killed after three days and the fundus, antrum, small intestine and pancreas were examined for neuroendocrine cells. Our study confirms that streptozotocin is cytotoxic towards beta-cells. In addition, it is cytotoxic towards neuroendocrine cells of the oxyntic mucosa of the stomach. This finding may have clinical significance and suggests that streptozotocin may be used in the treatment of gastric neuroendocrine tumors as well as insulinomas. PMID- 12772786 TI - Influence of cholecystitis state on pharmacological response to cholecystokinin of isolated human gallbladder with gallstones. AB - We studied the influence of the inflammatory state of the gallbladder with gallstones on its response to cholecystokinin (CCK). Responses to CCK were evaluated in isolated human gallbladder strips incubated with pharmacological antagonists. Gallbladders from patients with gallstones were classified as having mild and severe chronic cholecystitis. Healthy gallbladders were collected from liver donors. In donor gallbladders, the CCK contraction was abolished with the CCK-A receptor antagonist, L-364718, and significantly reduced by indomethacin. In gallbladders with gallstones, only mild cholecystitis showed a decreased contraction to CCK. In gallbladders with gallstones, no involvement of prostaglandins in the CCK response was observed. In severe cholecystitis, CCK contractile effect was reduced by the serotonin receptor antagonist methysergide. In healthy gallbladder, the contraction provoked by CCK is mediated by CCK-A receptors and modulated by prostaglandins. The presence of gallstones in the gallbladder is correlated with a loss of prostaglandins-modulated CCK contraction. However, the excessive release of serotonin in advanced cholecystitis normalizes the contraction to CCK, suggesting that the state of cholecystitis affects the pool of inflammatory mediators responsible for gallbladder CCK-altered motility. PMID- 12772788 TI - Primary sclerosing cholangitis associated with lupus nephritis: a rare association. PMID- 12772789 TI - Pancreas divisum and intraductal papillary mucinous tumor occurring simultanously in a patient presenting with recurrent acute pancreatitis. PMID- 12772790 TI - Clinical experience of hepatic hemangioma undergoing hepatic resection. AB - The indications for surgery on cavernous hemangiomas, the most common benign tumors of the liver, remain unclear. This study reviewed 43 patients with cavernous hemangioma of the liver who underwent hepatic resection from 1984 to 2000. Patients were divided into three groups based on the reasons for surgery. Group I comprised 13 patients whose lesions presented symptoms and dimensions that were the main indications for operation. Group II consisted of 28 patients diagnosed with malignant tumors or who displayed malignant growth that could not be ruled out preoperatively. Group III comprised 2 patients with tumors found incidentally at laparotomy for other malignancies. No surgical mortality related to hepatectomy was noted. Postoperative bile leak was found in 2 (morbidity rate: 4.7%). Patients were followed up from 6 months to 12 years. Thirteen residual tumors progressed in size. The clinical status or symptoms changed only slightly in 10 patients with recurrence. The results suggest that resection therapy is an effective indicator for patients with symptoms and a questionable diagnosis. Hepatic resection may and should be carried out with no mortality and minimal morbidity risks since the lesion is benign. PMID- 12772791 TI - Combination treatment of IFNalpha2b and ribavirin in patients with chronic hepatitis C and persistently normal ALTs. AB - Combination therapy of interferon-alpha2b and ribavirin was prospectively evaluated in 20 patients with chronic replicative hepatitis and persistently normal ALTs. Patients with normal ALTs on three or more occasions within 6 months received interferon-alpha2b 3 MU three times a week with ribavirin 1000-1200 mg everyday for 12 months and had a follow-up of 6 months. HCV genotype 1 was found in 16, and HCV genotype 2 or 3 in 4 patients. No patient experienced an ALT elevation during therapy. Ten of 20 patients (50%) cleared virus at the end of treatment. In an intent-to-treat analysis, a sustained virological response (SR) was achieved in 8 of 20 patients (40%). Nonresponse occurred in 5 patients. Relapse and breakthrough were seen in 2 patients each. Treatment was discontinued in 3 patients due to side effects. Interferon (IFN) ribavirin combination therapy is effective in patients with normal ALTs and appears superior to IFN monotherapy. PMID- 12772792 TI - Effects of daily, light and moderate-heavy ethanol exposure on extent of hepatic injury and recovery following toxin-induced acute hepatitis in rats. AB - Daily, light ethanol consumption enhances hepatic regeneration following 70% partial hepatectomy in rats. Whether such consumption has a beneficial effect on the outcome following toxin-induced acute hepatitis has yet to be determined. One hundred ten adult male Spragne-Dawlay rats (200-250 g) were randomized to receive daily gavages with ethanol 1.0 g/kg (light ethanol group), 3.0 g/kg (moderate heavy ethanol group), or an equal volume of tap water (controls). On day 30, a single injection of D-galactosamine hydrochloride (1.0 g/kg) (D-gal), a potent hepatotoxin that induces liver failure within 24-48 hr, was administered intraperitoneally. Gavages were discontinued and rats killed (N = 4-6/group) on days 1, 3, 5, 7, and 10 after D-gal. Serum AST, bilirubin, and liver histology served to document the extent of liver injury and [3H] thymidine incorporation into hepatic DNA: hepatic regenerative activity. Compared to controls, peak serum AST levels were significantly decreased in the light (-40%, P < 0.05) and increased in the moderate-heavy (+32%, P < 0.05) ethanol groups. Serum bilirubin levels approximately doubled in the light ethanol group while increasing sixfold in the moderate-heavy and control groups (P < 0.05). Histologic evidence of hepatic injury (graded 0-IV) was limited in the light ethanol group, intermediate in controls, and most extensive in the moderate-heavy ethanol group (P < 0.05). Despite less hepatic injury, hepatic regeneration was similar in the light ethanol group compared to controls and significantly impaired in the moderate heavy ethanol group (P < 0.01). In conclusion, the results of this study indicate that daily, light ethanol administration attenuates hepatic injury, improves hepatic function, and enhances hepatic regeneration following toxin-induced hepatitis in rats. PMID- 12772793 TI - Correlation between histopathological findings of the liver and IgA class antibodies to 2-oxo-acid dehydrogenase complex in primary biliary cirrhosis. AB - Although anti-mitochondrial antibody (AMA) is the characteristic serological feature of primary biliary cirrhosis (PBC), its pathogenetic role remains unclear. We tested sera from 72 Japanese patients with histologically confirmed PBC for AMA by indirect immunofluorescence, anti-pyruvate dehydrogenase complex (PDC) by enzyme inhibition assay, immunoglobulin (Ig) G class anti-PDC by ELISA, and IgG, IgM, and IgA class anti-2-oxo-acid dehydrogenase complex (2-OADC) by immunoblotting. Of the 72 sera, 60 (83%), 50 (69%), 42 (58%), and 71 (99%) were positive for AMA by immunofluorescence, enzyme inhibition assay, ELISA, and immunoblotting, respectively. There was no significant correlation between histological stages and AMA by immunofluorescence, PDC inhibitory antibodies by enzyme inhibition assay, IgG class anti-PDC antibodies by ELISA, or IgG and IgM class anti-2-OADC by immunoblotting. IgA class anti-2-OADC by immunoblotting was more frequent in stages 2-4 than in stage 1 (P = 0.0083). Of the IgA class anti-2 OADC, anti-PDC-E2 (74 kDa) and anti-E3BP (52 kDa) were more frequent in stages 2 4 than in stage 1 (P = 0.0253 and 0.0042, respectively). Further examination of histopathological findings in 53 of 72 liver biopsy specimens showed that IgA class anti-PDC-E2 and IgA class anti-E3BP were associated with bile duct loss, and IgA class anti-PDC-E2 was also associated with interface hepatitis and atypical ductular proliferation. IgA is known to be secreted into the bile through biliary epithelial cells, implying that IgA class anti-PDC-E2 and E3BP may have a specific pathogenetic role during their transport into the bile by binding to their target antigen(s) in biliary epithelial cells, and this may be followed by dysfunction and finally destruction of biliary epithelial cells. Our present results suggest that these autoantibodies against 2-OADC detected by immunoblotting may be associated with the pathogenesis and pathologic progression of PBC. PMID- 12772794 TI - Sertraline hepatotoxicity: a case report and review of the literature on selective serotonin reuptake inhibitor hepatotoxicity. AB - The causal role of sertraline in rare cases of liver failure in patients taking the drug has not been proven in a manner consistent with usually accepted standards. We describe an individual who developed clinically significant hepatitis while being treated with sertraline. This case is significant because it is the only one of which we are aware in which the diagnosis of sertraline hepatotoxicity was confirmed when inadvertent rechallenge with the medication resulted in recurrent hepatitis. We review this case and the general role of this widely prescribed class of drugs in causing hepatitis. PMID- 12772795 TI - Detection of Chlamydiae pneumoniae but not Helicobacter pylori DNA in atherosclerosis plaques. AB - Chronic infections have been associated with cardiovascular disease. We used bacterial culture, polymerase chain reaction (PCR), and immunohistochemical staining with anti-vacA and anticagA antibodies to search for Helicobacter pylori and Chlamydiae pneumoniae in atherosclerotic plaques obtained at endarterectomy. Serum IgG antibodies to H. pylori and C. pneumoniae were also determined. Thirty two patients were enrolled. Anti-H. pylori and anti-C. pneumoniae IgG were present in 72% and 81%, respectively. Culture and PCR for H. pylori of vessel walls and plaques were negative. Atherosclerotic plaque and normal vessel sections from H. pylori-negative and- positive patients showed reactivity with anti-vacA and anti-cagA antibodies. C. pneumoniae DNA was amplified in three atherosclerotic lesions. These findings suggest that the association between H. pylori infection and atherosclerosis does not result from continuing direct effects of H. pylori antigens in the vessel walls. Antigens within vessel atherosclerotic plaques cross-react with H. pylori virulence factors and could act as cofactors in determining instability for the atherosclerotic plaques. PMID- 12772796 TI - Utility of ambulatory 24-hour esophageal pH and motility monitoring in noncardiac chest pain: report of 90 patients and review of the literature. AB - It is unclear whether prolonged motility monitoring improves the diagnostic yield of standard esophageal tests in patients with noncardiac chest pain. Our aim was to assess the diagnostic value of ambulatory 24-hr pH and pressure monitoring in patients with noncardiac chest pain. Stationary manometry, edrophonium testing, and ambulatory pH and motility studies were performed in 90 consecutive patients with recurrent chest pain and normal coronary angiograms. Normality limits of ambulatory 24-hr motility were established in 30 healthy controls. The diagnoses of specific esophageal motility disorders (nutcracker esophagus and diffuse esophageal spasm) by stationary and ambulatory manometry were discordant in 48% of the patients. Edrophonium testing was positive in 9 patients, but correlated poorly with esophageal diagnoses. During ambulatory studies, 144 chest pain events occurred in 42 patients, and 72 (50%) were related to esophageal dysfunction. Strict temporal associations of events with esophageal dysfunction in relation to ambulatory 24-hr pH/motility scores permitted four patient categorizations: true positives (event-related and abnormal tests), N = 15; true negatives (event-unrelated and abnormal tests), N = 10; reduced esophageal pain threshold (event-related and normal tests), N = 4; and indeterminate origin (event-unrelated and normal tests), N = 13. Overall, 19 patients (21%) had a probable esophageal cause for chest pain (14 esophageal motility disorder, 4 acid reflux, 1 both). In conclusion, ambulatory manometry increases the diagnostic yield of standard esophageal testing in noncardiac chest pain, but the gain is small. Causes of chest pain other than high esophageal pressures and acid reflux must still be sought in most patients with chest pain of unknown origin after a negative cardiac work-up. PMID- 12772797 TI - Esophageal motility disorders in HIV patients. AB - Opportunistic esophageal infections (Candida, cytomegalovirus, herpes simplex virus) and idiophatic esophageal ulcerations are commonly found in HIV patients. However, motility disorders of the esophagus have seldom been investigated in this population. The aim of this prospective study was to determine the presence of motility disorders in HIV patients with esophageal symptoms (with or without associated lesions detected by endoscopy) and in HIV patients without esophageal symptoms and normal esophagoscopy. Eigthteen consecutive HIV patients (10 male, 8 female, ages 20-44 years, mean age 33.5; 8 HIV positive and 10 AIDS) were studied prospectively. Nine patients complained of esophageal symptoms, e.g, dysphagia/odynophagia (group 1) and 9 had symptoms not related to esophageal disease, such as diarrhea, abdominal pain, or gastrointestinal bleeding (group 2). All patients underwent upper endoscopy; mucosal biopsies were taken when macroscopic esophageal lesions were identified or when the patients were symptomatic even if the esophageal mucosa was normal. Esophageal manometry was performed in the 18 patients, using a 4-channel water-perfused system according to a standardized technique. Sixteen of the 18 patients (88.8%) had baseline manometric abnormalities. In group 1, 8/9 patients had esophageal motility disorders: nutcrackeresophagus in 1, hypertensive lower esophageal sphincter (LES) with incomplete relaxation in 2, nonspecific esophageal motility disorders (NEMD) in 3, diffuse esophageal spasm in 1, esophageal hypocontraction with low LES pressure in 1. Six of these 9 patients had lesions detected by endoscopy: CMV ulcers in 2, idiopathic ulcers in 1, candidiasis in 1, idiopathic ulcer + candidiasis in 1, nonspecific esophagitis in 1; and 3/9 had normal endoscopy and normal esophageal biopsies. In group 2, 8/9 patients had abnormal motility: hypertensive LES with incomplete relaxation in 1, nutcracker esophagus in 2, esophageal hypocontraction in 3, and NEMD in 2. All these patients had a normal esophageal mucosa at endoscopy. In conclusion, our findings suggest that HIV patients have esophageal motility disorders independent of esophageal symptoms and/or the presence of mucosal esophageal lesions. PMID- 12772798 TI - Nodular gastritis in adults is caused by Helicobacter pylori infection. AB - A close relationship exists between nodular gastritis and Helicobacter pylori infection in children. The pathogenesis and optimal management of nodular gastritis in adults, however, are unclear. This study describes the clinicopathologic features of nodular gastritis in adults and correlates treatment with outcome. Of 97,262 adult patients who underwent endoscopy, 187 (0.19%) were diagnosed with nodular gastritis, 151 (81%) of whom had dyspepsia. Nodular gastritis predominantly affects young women (49 men and 138 women, mean age, 32.6 years). All 134 patients tested for Helicobacter pylori infection were infected, and 65/66 (98%) had inflammation of both the antrum and the corpus. Twenty-five (13%) had associated lesions (peptic ulcers or cancer). Dyspepsia improved after eradication of Helicobacter pylori infection, but did not improve spontaneously. Nodular gastritis in adults is caused by Helicobacter pylori infection and shows a predilection for females and young adults. Helicobacter pylori eradication decreases symptoms and reduces the risk of peptic ulcers and possibly gastric cancer. PMID- 12772799 TI - Decreased gastric bacterial killing and up-regulation of protective genes in small intestine in gastrin-deficient mouse. AB - Gastrin regulates gastric acid secretion, believed to be primarily responsible for killing ingested microbes. We examined gastric killing of gavaged E. coli in gastrin-deficient mice, which have decreased gastric acid production. Additionally, the expression of intestinal genes involved in epithelial protection were analyzed: the mucus layer glycoprotein muclin, the polymeric Ig receptor, trefoil factor 3, and small proline-rich protein 2a (sprr2a). Gastric pH was 2.5 pH units greater in gastrin-deficient mice, and E. coli survival was increased greater than 20-fold at 10 min after gavage compared to control. Muclin and sprr2a gene expression were significantly increased (2.0- and 2.6-fold) in the intestine, and antibiotic treatment reversed these effects. In conclusion, reduced gastric acid secretion results in increased survival of ingested microorganisms in gastrin-deficient mice. Bacterial survival is associated with increased expression of muclin and sprr2a in the intestine, indicating that these genes play protective roles in the intestine. PMID- 12772800 TI - Preventive effect of zaprinast and 3-isobutyl, 1-methylxanthine (phosphodiesterase inhibitors) on gastric injury induced by nonsteroidal antiinflammatory drugs in rats. AB - Cyclic GMP plays an important role in maintaining homeostasis in the gastric mucosa. NSAIDs damage the mucosa by mechanisms that may be mediated by alterations in the intragastric concentration of cyclic GMP. To test this hypothesis we studied the effects of the oral administration of acetylsalicylic acid (100, 300, and 500 mg/kg), piroxicam (5, 10, and 20 mg/kg) and sodium diclofenac (10, 25, 50, and 100 mg/kg), and of their interaction with zaprinast (5 mg/kg) and IBMX (10 mg/kg), on intragastric concentrations of cyclic GMP and the gastric erosive index in rats. All determinations were done 3 hr after the NSAID was given. All NSAIDs induced dose-dependent decreases in mucosal concentrations of cyclic GMP, which correlated inversely with the surface area showing mucosal injury. In contrast, cyclic GMP concentrations remained normal, and no intragastric damage was seen in rats given zaprinast (cyclic GMP-specific phosphodiesterase inhibitor) or IBMX (non-specific phosphodiesterase inhibitor) or in combination with NSAIDs. These findings are in line with the hypothesis that cyclic GMP is involved in the biochemical mechanisms of NSAID-induced gastric injury. PMID- 12772801 TI - Crohn's disease with Parkinsonism due to long-term total parenteral nutrition. PMID- 12772802 TI - Isolation of human small intestinal brush border membranes using polyethylene glycol and effect of exposure to various oxidants in vitro. AB - This study presents a method of brush border membrane (BBM) preparation from the human small intestine using polyethylene glycol (PEG) precipitation and also looks at the effect of in vitro oxidant exposure on structural and functional alterations in the membrane. Isolated BBM were relatively pure as judged by 10- to 14-fold enrichment of marker enzymes with less than 1% contamination by other subcellular organelles. These membranes showed uphill transport of glucose and lipid analysis showed a cholesterol-phospholipid (C/P) ratio of 1.19. Isolated BBM were found to be susceptible to superoxide generated by xanthine oxidase (XO), resulting in lipid and protein oxidation along with altered glucose uptake. Superoxide exposure also resulted in phospholipid alterations, especially generation of lyso phospholipids. These changes were prevented by inhibiting XO by allopurinol or scavenging superoxide by superoxide dismutase (SOD). Other oxidants studied did not have significant affect on these membranes. These studies suggest that PEG can be used for preparation of BBM from the human small intestine and these membranes undergo structural and functional alterations on exposure to superoxide. PMID- 12772803 TI - Outpatient treatment of moderately severe active ulcerative colitis with pulsed steroid therapy and conventional steroid therapy. AB - Pulsed steroid therapy may induce rapid remission in patients with moderately severe ulcerative colitis in outpatient clinics. A total of 19 patients with moderately severe active ulcerative colitis who refused hospitalization were treated between October 1999 and September 2001 in the outpatient clinic. Patients were treated with either conventional oral steroid therapy or intravenous pulsed steroid therapy followed by conventional oral steroid therapy. Eight patients received conventional steroid therapy and 11 patients received pulsed steroid therapy followed by conventional steroid therapy. The efficacies of the two types of steroid therapy were equal, but patients with active colitis responded more quickly to pulsed steroid therapy than to conventional steroid therapy. No serious adverse effects were observed. Moderately severe colitis can be safely treated with either conventional or pulsed steroid therapy in the outpatient clinic, but pulsed steroid therapy may induce clinical remission more quickly than conventional steroid therapy. PMID- 12772804 TI - Scanning electron microscopic study of cold-stored small bowel: comparison of Euro-Collins and Lactated Ringer's solutions. AB - The increasing use of the small bowel for transplantation necessitates the development of optimum preservation techniques. The aim of our study was to investigate time-related morphological changes of the rat small bowel during preservation of the organ in hypothermic Euro-Collins and Lactated Ringer's solutions. It is the only study comparing these widely used preservation solutions with scanning electron microscopy, which is especially suitable to show the intestinal mucosa, one of the tissues of the intestine most susceptible to ischemia. Small bowels were perfused with either Euro-Collins, Lactated Ringer's or physiological saline solution and placed in respective preservation solutions at 4 degrees C for 0, 3, 6 and 12 hours. Biopsy samples were prepared for examination at the light and scanning electron microscopic levels. According to the results of this study we conclude that, even Lactated Ringer's solution showed better tissue preservation than Euro-Collins solution, both preservation solutions are suitable for short term preservation of the small bowel and that further ameliorations of the preservation solutions and/or techniques are required for long term preservation. PMID- 12772806 TI - Foreign body removal through appendicostomy. PMID- 12772807 TI - Recurrent mantle cell lymphoma after allogeneic bone marrow transplantation presenting as isolated rectal mass. PMID- 12772805 TI - Eosinophilic gastroenteritis with ascites: a case report and review of the literature. PMID- 12772808 TI - A study of transition: the new nurse graduate at 3 months. AB - BACKGROUND: Nursing shortages, increased patient acuity, and early discharges have increased demands on newly graduated nurses. At the same time, financial constraints have curtailed orientation programs. METHOD: Open-ended interviews were conducted with 11 nurses employed in acute care settings at 3 months following graduation. RESULTS: New nurses defined their work as a set of skills and attended to procedural aspects of care. Most lacked the capacity for helpful communications with patients and families. They relied heavily on routines learned from experienced nurses. CONCLUSION: At 3 months, new graduates are apprehensive about their work. Approaches to nursing are largely procedural and guided by the routines of senior colleagues. PMID- 12772809 TI - A strategy for involving on-campus and distance students in a nursing research course. AB - BACKGROUND: Teaching research at the undergraduate level can be a challenge. This is particularly true for distance education courses, in which students often feel isolated from each other and from faculty. Faculty teaching on-campus and distance education research courses designed a student research project and a method for bringing distance and on-campus students together to present their research findings. METHOD: By actively engaging students in the research process and providing an on-campus research day for the presentation of student posters (Year 1) and research papers (Year 2), course faculty were able to achieve their two goals of directly involving students in the nursing research process and creating a greater student sense of belonging within the college learning community. RESULTS: Statistical analysis of an evaluation survey among on-site and off-site registered nurse to bachelor of science in nursing students indicated both groups ranked the Year 1 poster research day as good to very good. Students attending the Year 2 research day, where they presented papers, ranked the day as good. Statistical analysis indicated a significant difference between the two research days, with a clear student preference for posters. CONCLUSION: Students valued their active participation in the research process and the opportunity to get together and present their work. However, students clearly preferred a poster presentation format to presenting their findings as a research paper. PMID- 12772810 TI - Start out: building healthcare careers for minority teenagers. AB - BACKGROUND: The healthcare industry is currently experiencing a severe shortage of nurses and other allied healthcare professionals. To compound this crisis, the current workforce does not mirror the growing linguistic and cultural diversity of the population needing health care. This article describes a community partnership approach to creating educational and career opportunities in nursing and other health sciences for young, multicultural, bilingual teenagers. METHOD: Several community organizations joined together to design and implement an 8-week summer program entitled "Start Out," which served to integrate life planning, mentorship, nursing assistant training, and college application assistance while providing summer salary stipends and work scholarship opportunities. Twenty-seven students participated in the program. RESULTS: Twenty-four bilingual, economically disadvantaged teenagers 16 to 19 years old, from the greater Seattle metropolitan area, completed the first session of this innovative program and passed the certified nursing assistant examination. CONCLUSION: Culturally focused assistance programs may provide an avenue to meet the current nursing shortage and provide healthcare workers who can be sensitive to the cultural and linguistic needs of the growing ethnic population. PMID- 12772811 TI - Nurses' needs for education on cancer and end-of-life care. AB - BACKGROUND: Changes in cancer and end-of-life care require frequent assessment of educational needs of nurses. METHODS: The Nurse Oncology Education Program surveyed a random sample of Texas registered nurses about their continuing education practices, level of knowledge, and educational needs. RESULTS: The 352 nurses responding to the survey primarily obtained continuing education from workshops, inservice education, and independent studies citing cost, location, content, and length of course as influencing factors. Their cancer educational needs included clinical trials, genetics, complementary therapies, and pain management. Nurses' perceptions of end-of-life needs were for physical needs, "what to expect" concerns, and transition to palliative care. CONCLUSION: These findings provide specific direction for future continuing education programs about cancer and end-of-life issues for nurses. PMID- 12772812 TI - The development of an educational workshop on complementary and alternative medicine: what every nurse should know. AB - As complementary and alternative medicine (CAM) is becoming more accepted in Western culture, healthcare consumers are choosing CAM as an adjunct to conventional healthcare practices. A variety of cultural backgrounds contributes to the need to advance knowledge and evaluate outcomes of healthcare practices related to CAM. The issues presented in this article provide critical information for nurses and other healthcare providers to integrate conventional medicine with CAM practices to improve healthcare outcomes for patients who use CAM. PMID- 12772813 TI - Multicenter, open-label evaluation of hyperemia associated with use of bimatoprost in adults with open-angle glaucoma or ocular hypertension. AB - This month-long multicenter, open-label study evaluated the onset and progression of hyperemia associated with bimatoprost 0.03% once daily in 39 patients with open-angle glaucoma or ocular hypertension. Current glaucoma medication(s) was either replaced with or augmented by bimatoprost. Previous users of bimatoprost were excluded. Primary outcome measures were mean hyperemia scale scores (ciliary, conjunctival, and episcleral, graded on a seven-point scale) and incidence of hyperemia. Secondary outcome measures were fluorescein staining, patient assessment of ocular redness (take-home diary), and patient and investigator evaluations. Patients were asked how troubled they were by their ocular redness. Investigators were asked if they would continue the patient on bimatoprost despite the hyperemia. Overall, the frequency and severity of hyperemia peaked approximately 1 day after the first instillation of bimatoprost and decreased consistently throughout the study, returning to near-baseline levels by day 28. At day 1, 84.6% of patients were hardly troubled or not troubled by their ocular redness; only 15.4% were somewhat or moderately troubled. Investigators indicated that they would continue bimatoprost therapy in 92.3% of the patients. Hyperemia did not represent a significant safety concern. PMID- 12772814 TI - Sequential chemoradiotherapy versus radiotherapy in the management of locally advanced non-small-cell lung cancer. AB - This prospective randomized trial compared neoadjuvant chemotherapy followed by local-regional radiotherapy (study group) with radiotherapy alone (control group) in 506 patients with locally advanced non-small-cell lung cancer. The study group received three cycles of cisplatin, ifosfamide with Mesna, and mitomycin C at 21 day intervals. Radiotherapy was delivered up to a dose of 60 Gy. Evaluable patients numbered 460. In the study group, 228 patients were assessed for tumor response after chemotherapy: 13 (5.7%) had a complete response; 103 (43.2%), a partial response; 48 (21%) showed no change; and 74 patients (31.1%) had progressive disease. These patients then received chest irradiation, which was well tolerated. On completion of radiotherapy, 16.2% of the study group and 6% of the control group had a complete response. No change was seen in 10.1% of study patients and 15.5% of control patients. Respective rates for disease progression were 27.6% and 42.3%. Actuarial 2-year survival was 20% in the study group and 7.4% in the control group. PMID- 12772815 TI - Brimonidine Purite and bimatoprost compared with timolol and latanoprost in patients with glaucoma and ocular hypertension. AB - This 3-month multicenter, investigator-masked, parallel-group study compared brimonidine Purite and bimatoprost (brimP/bim) with timolol gel-forming solution and latanoprost (tim/latan) in 28 patients with open-angle glaucoma or ocular hypertension. IOP was measured at baseline and 2 hours after morning instillation at weeks 2, 4, and 12. The primary outcome measure was the mean IOP reduction from baseline. Secondary measures were the percentage of patients in each group who achieved specific low target pressures and the incidence of adverse events. Mean IOP at baseline was 24.8 mm Hg in each group. At follow-up visits, mean reductions from baseline ranged from 8.5 to 9.0 mm Hg with brimP/bim and from 7.5 to 7.7 mm Hg with tim/latan. More patients achieved low target pressures with brimP/bim. At week 12, 69.2% of brimP/bim patients and 27.3% of tim/latan patients had IOPs of 16 mm Hg or lower (P = .024). Both regimens were well tolerated, and adverse events were infrequent. The combination of brimonidine Purite and bimatoprost was well tolerated and at least as effective as Timoptic XE and latanoprost in reducing IOP. More patients achieved low target IOPs with brimonidine Purite and bimatoprost than with Timoptic-XE and latanoprost. A larger study is needed to confirm these results. PMID- 12772816 TI - Two mast cell stabilizers, pemirolast potassium 0.1% and nedocromil sodium 2%, in the treatment of seasonal allergic conjunctivitis: a comparative study. AB - This randomized, double-masked, active-control, parallel-group trial compared the mast cell stabilizers pemirolast potassium 0.1% and nedocromil sodium 2% in the treatment of seasonal allergic conjunctivitis. Pemirolast is currently indicated for four-times-daily administration, nedocromil, for twice-daily dosing. Both ophthalmic solutions were instilled bilaterally twice a day for 8 weeks. The study involved four office visits and two telephone contacts. Participants evaluated their symptoms daily in take-home diaries (itching was the primary efficacy variable) and completed questionnaires to assess comfort. Of a total enrollment of 80, 78 patients completed the study. No significant differences were found between pemirolast and nedocromil on any signs or symptoms of allergic conjunctivitis (redness, chemosis, itching, eyelid swelling). At each visit, pemirolast was rated significantly more comfortable than nedocromil. A significantly higher percentage of the pemirolast group experienced no signs or symptoms at work or school (58% vs 28%; P = .005). The number of adverse events did not differ significantly between groups. Twice-daily administration of pemirolast potassium was as efficacious and safe as twice-daily nedocromil sodium in the 8-week treatment of ragweed allergic conjunctivitis and was superior to nedocromil in comfort. Increased comfort with pemirolast may increase patient satisfaction and compliance with therapy. PMID- 12772817 TI - Efficacy of a herbal preparation in patients with functional dyspepsia: a meta analysis of double-blind, randomized, clinical trials. AB - A meta-analysis was performed of double-blind, randomized clinical studies that evaluated the efficacy of the herbal preparation Iberogast in patients with functional dyspepsia. All studies had the same duration and used the same dosage of active treatment and the same primary outcome measure, a dyspepsia-specific gastrointestinal symptom score. Of the 592 trial participants, 196 were treated with Iberogast and 192 with placebo or cisapride (positive control). The individual studies all showed a substantial improvement of symptoms with Iberogast but varying results regarding its statistically significant superiority to placebo. The meta-analysis of all studies, however, demonstrated a clear, highly significant overall therapeutic effect of Iberogast in the treatment of functional dyspepsia. Tolerability of the preparation was excellent. PMID- 12772818 TI - Effects of soy and other natural products on LDL:HDL ratio and other lipid parameters: a literature review. AB - Abnormal lipid levels contribute significantly to the risk of coronary heart disease, a major cardiovascular disease and a serious health problem. Various dietary and pharmacologic treatments have been devised to reduce elevated blood cholesterol levels. Soy protein, soluble fiber, and plant sterol/ester-containing margarines are promising new food-component candidates that may help to realize this goal. Of particular interest in this context is the LDL:HDL ratio, a strong predictor of cardiac events. This report is a review of more than 50 recent trials to determine how such dietary components and garlic affect the LDL:HDL ratio and other lipid parameters. Consumption of new soy products containing high, fixed levels of isoflavones, cotyledon soy fiber, and soy phospholipids (Abaco and Abalon) significantly reduced the LDL:HDL ratio by up to 27%. Soluble dietary fibers such as psyllium and beta glucan from oat bran had a variable effect on LDL-cholesterol levels in the studies analyzed. Plant sterol esters, when consumed in margarines, lowered the LDL:HDL ratio by up to 22%. On average, Abacor and Abalon reduced the LDL:HDL ratio by 20%, LDL cholesterol by 15%, total cholesterol by 10%, and triglycerides by 6%, and increased HDL cholesterol by 5%. The new soy-based supplements may therefore play a valuable role in reducing cardiovascular risk. PMID- 12772819 TI - Ecohydrology--why hydrologists should care. PMID- 12772820 TI - Perspectives on Turkish ground water resources. PMID- 12772821 TI - Putting shared knowledge to work. PMID- 12772822 TI - A fresh water odyssey: some observations on the global resource. AB - Appreciable increases in the human population are expected to continue in the next 50 to 100 years. This population will require additional water for nonfungible (nonexchangeable) uses such as irrigated agriculture, livestock watering, domestic supply, and ecosystem support. Because most of the world's easily captured water is already identified and allocated, society must improve efficiency, change the present allocations, and/or develop new sources to meet the expected demands. As the global economy expands, apparently unrelated changes in policy or technology may have large, unexpected consequences for water resources. Foreshadowing these changes in stress on these resources will be the result of nonlinear thinking. Whereas policy cannot create new water, it can provide strategies to promote more efficient use of present water and foster an environment in which important technological improvements can be made. This manuscript provides a brief overview of some of the physical and policy considerations relating to fresh water resources. PMID- 12772823 TI - A digital procedure for ground water recharge and discharge pattern recognition and rate estimation. AB - A digital procedure to estimate recharge/discharge rates that requires relatively short preparation time and uses readily available data was applied to a setting in central Wisconsin. The method requires only measurements of the water table, fluxes such as stream baseflows, bottom of the system, and hydraulic conductivity to delineate approximate recharge/discharge zones and to estimate rates. The method uses interpolation of the water table surface, recharge/discharge mapping, pattern recognition, and a parameter estimation model. The surface interpolator used is based on the theory of radial basis functions with thin-plate splines. The recharge/discharge mapping is based on a mass-balance calculation performed using MODFLOW. The results of the recharge/discharge mapping are critically dependent on the accuracy of the water table interpolation and the accuracy and number of water table measurements. The recharge pattern recognition is performed with the help of a graphical user interface (GUI) program based on several algorithms used in image processing. Pattern recognition is needed to identify the recharge/discharge zonations and zone the results of the mapping method. The parameter estimation program UCODE calculates the parameter values that provide a best fit between simulated heads and flows and calibration head-and-flow targets. A model of the Buena Vista Ground Water Basin in the Central Sand Plains of Wisconsin is used to demonstrate the procedure. PMID- 12772824 TI - A spreadsheet method of estimating best-fit hydraulic gradients using head data from multiple wells. AB - Hydraulic gradients from planar water tables, or piezometric surfaces, and horizontal flow regimes can be quickly and conveniently calculated from data sets involving numerous wells. The matrix-solving functions of a modem spreadsheet program (Excel) were used to determine the equation of a water-table plane, Ax + By + Cz - D = 0, and the equation coefficients were then used to determine the magnitude of the hydraulic gradient, according to gradient = square root of A2 + B2/C2, and its direction, according to alpha = arctan B/A, where alpha is the angle measured from the x-axis. A pre-prepared Excel file constructed to handle data from up to 20 wells at once is available for free downloading at www.geo.ku.edu/hydro/KUHydro.html. PMID- 12772825 TI - The permeability of the Elkhorn fault zone, South Park, Colorado. AB - The purposes of this study are to use both field and modeling approaches to characterize the permeability of a fault and to assess the role of the fault on regional ground water flow. The study subject is the Elkhorn fault, a low-angle reverse fault that brings Precambrian crystalline rocks over the sediments of Colorado's South Park Basin. The fault is hypothesized to act as a low permeability barrier to flow, restricting interaction between the crystalline aquifer and the basin sediments. To test this hypothesis and to better predict the permeability structure of the fault, we synthesized geologic data to create a geologic model of the fault, conducted aquifer tests to estimate the hydrogeologic properties of the fault zone, and used ground water modeling to test the influence of a range of hydraulic properties for the fault zone on ground water flow in the region. Our study suggests that the fault is a low permeability feature. Estimated heads are best matched to observations by modeling the fault as a 10-foot-thick interval of low-permeability fault gouge. Steady-state flow models show that much of the flow in the study area is topographically driven near land surface. Flow rates decrease with depth in the aquifers. In the footwall, ground water moves updip in the Michigan-San Isabel syncline to discharge in the South Park Basin. In the hanging wall, ground water moves east to a regional ground water divide. Sensitivity analyses indicate that hydraulic heads are most sensitive to changes in hydraulic conductivity and recharge. PMID- 12772826 TI - Flow dimension as an indicator of hydraulic behavior in site characterization of fractured rock. AB - We examine the possibility of using the flow dimension identified from constant pressure injection tests as a tool for characterizing the hydraulic conditions of fractured media. The data comes from a low-conductivity crystalline rock site, from depths of up to 450 m, and is obtained with 2 m and 10 m measurement scales. In the analysis, the general solution for n-dimensional flow by Barker (1988) is applied. The results show that the most prominent characteristics of the medium can be identified; that is, linear and sublinear flow dimensions as distinguished from dimensions higher than two. In many cases, however, there is significant difficulty in distinguishing the dimensions n = 2, 2.5, and 3 from each other. This is usually because of the experimental difficulties in achieving the ideal conditions required by the theory during the early part of the experiment. In such cases, a full flow curve is not available for the type-curve fitting. In the nonunique cases the higher dimensions typically correspond to higher, sometimes unrealistically high, values of specific storage and to the less reliable and less representative early part of the experiment. Therefore, most of the dimensions in categories n = 3 can be excluded, thus leaving the majority observations in the categories of n = 2 and n = 2-2.5. The dominance of dimension n = 2 is more pronounced for data related to fracture zones in comparison to that related to "average" rock, in particular in the 2 m scale data. The proportion of low (n < 1.5) flow dimensions is small, but for the 10 m scale data it is relatively higher at greater depths and corresponds to lower conductivities. For the smaller 2 m scale data, the low dimensions are not linked to greater depths or systematically smaller conductivities, giving preliminary indication of different flow dimension behavior for the two different scales. PMID- 12772827 TI - MODFLOW 2000 head uncertainty, a first-order second moment method. AB - A computationally efficient method to estimate the variance and covariance in piezometric head results computed through MODFLOW 2000 using a first-order second moment (FOSM) approach is presented. This methodology employs a first-order Taylor series expansion to combine model sensitivity with uncertainty in geologic data. MODFLOW 2000 is used to calculate both the ground water head and the sensitivity of head to changes in input data. From a limited number of samples, geologic data are extrapolated and their associated uncertainties are computed through a conditional probability calculation. Combining the spatially related sensitivity and input uncertainty produces the variance-covariance matrix, the diagonal of which is used to yield the standard deviation in MODFLOW 2000 head. The variance in piezometric head can be used for calibrating the model, estimating confidence intervals, directing exploration, and evaluating the reliability of a design. A case study illustrates the approach, where aquifer transmissivity is the spatially related uncertain geologic input data. The FOSM methodology is shown to be applicable for calculating output uncertainty for (1) spatially related input and output data, and (2) multiple input parameters (transmissivity and recharge). PMID- 12772828 TI - Analytical model for computing residence times near a pumping well. AB - An analytical solution for calculating the residence time of fluid flowing toward a pumping well in an unconfined aquifer has been developed. The analytical solution was derived based on a radial, steady-state, Dupuit-Forchheimer flow model. The resulting integral expression involved computing the imaginary error function, for which a simple series expansion is proposed. The validity of the analytical expression is demonstrated by testing its results against numerical results for an example problem. The analytical solution compared favorably with the numerical approximation. PMID- 12772829 TI - MAROS: a decision support system for optimizing monitoring plans. AB - The Monitoring and Remediation Optimization System (MAROS), a decision-support software, was developed to assist in formulating cost-effective ground water long term monitoring plans. MAROS optimizes an existing ground water monitoring program using both temporal and spatial data analyses to determine the general monitoring system category and the locations and frequency of sampling for future compliance monitoring at the site. The objective of the MAROS optimization is to minimize monitoring locations in the sampling network and reduce sampling frequency without significant loss of information, ensuring adequate future characterization of the contaminant plume. The interpretive trend analysis approach recommends the general monitoring system category for a site based on plume stability and site-specific hydrogeologic information. Plume stability is characterized using primary lines of evidence (i.e., Mann-Kendall analysis and linear regression analysis) based on concentration trends, and secondary lines of evidence based on modeling results and empirical data. The sampling optimization approach, consisting of a two-dimensional spatial sampling reduction method (Delaunay method) and a temporal sampling analysis method (Modified CES method), provides detailed sampling location and frequency results. The Delaunay method is designed to identify and eliminate redundant sampling locations without causing significant information loss in characterizing the plume. The Modified CES method determines the optimal sampling frequency for a sampling location based on the direction, magnitude, and uncertainty in its concentration trend. MAROS addresses a variety of ground water contaminants (fuels, solvents, and metals), allows import of various data formats, and is designed for continual modification of long-term monitoring plans as the plume or site conditions change over time. PMID- 12772830 TI - Estimating ground water discharge by hydrograph separation. AB - Iron Mountain is located in the West Shasta Mining District in California. An investigation of the generation of acid rock drainage and metals loading to Boulder Creek at Iron Mountain was conducted. As part of that investigation, a hydrograph separation technique was used to determine the contribution of ground water to total flow in Boulder Creek. During high-flow storm events in the winter months, peak flow in Boulder Creek can exceed 22.7 m3/sec, and comprises surface runoff, interflow, and ground water discharge. A hydrograph separation technique was used to estimate ground water discharge into Boulder Creek during high-flow conditions. Total ground water discharge to the creek approaches 0.31 m3/sec during the high-flow season. The hydrograph separation technique combined with an extensive field data set provided reasonable estimates of ground water discharge. These estimates are useful for other investigations, such as determining a corresponding metals load from the metal-rich ground water found at Iron Mountain and thus contributing to remedial alternatives. PMID- 12772831 TI - Transport of three herbicides in ground water at Twin Lake test site, Chalk River, Ontario, Canada. AB - A field tracer test performed under natural flow conditions at the Twin Lake test site, Chalk River Laboratories of the Atomic Energy of Canada Ltd. in Chalk River, Ontario, Canada, using tritium and three herbicides (Chlortoluron, Terbuthylazine, and Pendimethalin) was interpreted using the dispersion equation with a combined reaction model. The reaction model couples an instantaneous equilibrium reaction governed by a linear adsorption isotherm with a reversible or irreversible kinetic reaction of the first order, and decay. An improved interpretation method consists of a simultaneous fitting of theoretical concentration and mass-recovery curves to the experimental data, which leads to a more reliable determining of reaction models and improves the accuracy of fitting. Tritium served as the reference tracer to determine the flow velocity, dispersivity, and the recovery of the herbicides. Chlortoluron was slightly delayed by equilibrium exchange with strongly reduced concentration due to an irreversible kinetic reaction and/or decay. Terbuthilazine was slightly delayed by equilibrium exchange, with strongly reduced concentration due to a reversible kinetic reaction with some influence of decay. A strong equilibrium reaction and a strong reversible kinetic reaction without degradation governed the transport of Pendimethalin, reducing considerably its concentration. The results obtained show that simulations based only on Kd and decay constant, especially if these parameters are found in the laboratory, may considerably differ from those performed with reaction parameters determined in properly performed field tests. The dominant reaction types, and the values of parameters found in the study, supply useful information on the transport of the investigated herbicides in sandy aquifers under natural flow conditions. PMID- 12772832 TI - Revisit of well function approximation and an easy graphical curve matching technique for Theis' solution. AB - Some approximations of Theis' solution are reviewed and compared with the numerical evaluation of the exponential integral by means of a short FORTRAN code. A graphical technique of Theis' curve matching is proposed to make this procedure as easy as plotting a graph. PMID- 12772833 TI - "Stream-aquifer interactions: evaluation of depletion volume and residual effects from ground water pumping," by Xunhong Chen and Longcang Shu, May-June 2002 issue, v. 40, no. 3: 284-290. PMID- 12772834 TI - Jacob Bear: an autobiography. PMID- 12772835 TI - Stockpiled forage or limit-fed corn as alternatives to hay for gestating and lactating beef cows. AB - In Exp. 1, 31, 24, and 17 mature, pregnant Simmental x Angus cows (initial BW = 662.0 +/- 10.4 kg) in each of 3 yr were used to determine the efficacy of stockpiled orchardgrass, limit-fed corn, or ad libitum hay for maintaining cows in mid- to late gestation, respectively. In Exp. 2, 24 mature, pregnant crossbred cows (initial BW = 677.7 +/- 9.4 kg) per treatment in each of 3 yr were used to determine the efficacy of stockpiled orchardgrass, limit-fed corn, or ad libitum hay for maintaining cows in late gestation and early lactation, respectively. Each year, cows were assigned to treatment by BW. From November to February or from January to April, respectively, nutritional needs for mid- to late gestation (Exp. 1) or late gestation and early lactation (Exp. 2) were met either by 1) rotating cows on approximately 15.2 or 21.7 ha of predominantly orchardgrass pasture, set aside and fertilized in late August, 2) limit-feeding approximately 5.8 kg of whole shelled corn, 1.1 kg of a pelleted supplement, and 1.2 kg of hay daily, or 3) ad libitum feeding of round-baled hay. During extreme weather conditions, cows grazing stockpiled orchardgrass were limit-fed a grain-based diet. Postcalving weight (P < 0.10) was greatest for limit-fed cows in Exp. 1 and lowest for cows grazed on stockpiled orchardgrass; cows given ad libitum access to hay were intermediate in weight and did not differ from cows limit-fed or cows grazed on stockpiled orchardgrass (641.8, 657.4, and 634.0 kg, respectively). Calving date, calf birth and weaning weight, and conception rate did not differ among treatments (P > 0.15) in Exp. 1. In Exp. 2, weight at weaning did not differ among treatments (P > 0.17); however, postcalving weight (P < 0.01) was greatest for cows given ad libitum access to hay, intermediate for limit-fed cows, and lowest for cows grazed on stockpiled orchardgrass (674.8, 652.4, and 624.5 kg). Body condition score at any time point did not differ among treatments (P > 0.38), nor did calving date, calf birth and weaning weights, and conception rate (P > 0.12). Because of the few differences in cow performance, selection of energy sources for beef cows can be made based on economics. The cost to feed a cow hay in early to mid-gestation was nearly double that of limit-feeding the corn-based diet or grazing stockpiled orchardgrass. Because of lower quality pastures, the cost to graze cows on stockpiled orchardgrass during late gestation and early lactation was not as cost effective as limit feeding a corn-based diet. PMID- 12772836 TI - Development of a model to describe the compositional growth and dietary lysine requirements of pigs fed ractopamine. AB - The objective of this research was to use recent ractopamine research data to develop an updated mathematical model to describe the daily compositional growth of pigs fed ractopamine. Mean increases of 18.2, 23.1, and 25.0% for daily protein accretion were assumed for 5, 10, and 20 ppm of ractopamine for an overall gain of 40 kg of BW gain during the feeding period. The relative effect of ractopamine described the rapid increase and subsequent decrease in the effect of ractopamine as a function of BW gain or days on test and ractopamine concentration (RC, ppm). The reduction in ME intake produced by ractopamine was described as 0.036 x (RC/20)(0.7) multiplied by the ME intake for the first 20 kg of BW gain, and then increasing to 0.078 x (RC/20)(0.7) at 40 kg of BW gain feeding period. The ratio of fat-free muscle gain to protein accretion increased by 14 to 16% with the feeding of ractopamine, depending on the dietary lysine/essential AA levels. The ratio of carcass fat gain to empty body lipid gain was increased when lysine and essential AA requirements were met. Daily protein accretion and fat-free lean growth were described as functions of dietary lysine/essential AA intakes. The percentage of lysine in protein accretion increased with the feeding of ractopamine from 6.80 to 7.15%, depending on ractopamine concentration. Equations predicting carcass measurements, such as fat and longissimus muscle depths from carcass weight and composition, were modified to incorporate prediction biases produced by ractopamine. For the four concentrations of ractopamine (0, 5, 10, and 20 ppm, respectively) during a 78 to 110 kg of BW feeding period, the model predicted performance levels for ADG (1.03, 1.15, 1.16, and 1.16 kg/d), gain:feed (kg of ADG/kg of ADFI; 0.360, 0.401, 0.412, and 0.425), dressing percentage (75.1, 76.0, 76.3, and 76.4), percentage fat-free lean (48.7, 51.0, 51.5, and 52.2), longissimus muscle area (38.8,41.8,42.5, and 43.5 cm2), 10th-rib fat depth (22.1, 19.8, 19.3, and 18.7 mm), and fat-free lean gain (321, 446, 467, and 495 g/d), comparable to recent research data. The model allows the effect of ractopamine to be added to farm specific pig growth curves. It can be used to evaluate ways to optimize the use of ractopamine, including duration of ractopamine feeding, concentration of ractopamine, and dietary lysine concentration. PMID- 12772838 TI - The growth performance of the progeny of two swine sire lines reared under different floor space allowances. AB - A total of 736 pigs was used in a study with a 2 x 2 x 2 factorial arrangement to investigate the effects of and interactions between sire line (Line A vs. B), floor space (unrestricted vs. restricted), and gender (barrow vs. gilt) on growth performance, BW, and protein and estimated lipid accretion curves from 40 to 120 kg of BW. Pigs were by eight Line-A and nine Line-B sires mated with PIC C22 dams. Line A was of Pietrain ancestry and Line B was a synthetic line. The unrestricted floor space treatment consisted of small groups (four pigs) with 0.93 m2/pig of floor space for the entire grow-finish period. Pigs in the restricted floor space were in larger groups (12 pigs) with 0.37 and 0.56 m2/pig of floor space for the grower and finisher phases, respectively. Pigs were given ad libitum access to a three-phase dietary program, and one and three nipple waterers were available in the groups of 4 and 12 pigs, respectively. No sire line x floor space interactions were found for any of the traits measured. Line A pigs grew more slowly (50 g/d, P < 0.05), took longer (4.1 d, P < 0.05) to reach harvest weight (120.3 kg), and had similar feed intakes, but a lower gain:feed ratio (2.8%, P < 0.05) than Line B pigs. Line A pigs had greater longissimus muscle depth (P < 0.05) and estimated protein accretion rate (P < 0.05) than Line B pigs, but Line A and Line B pigs had a similar estimated percentage of lipid free soft tissue. Pigs reared in the restricted floor space grew more slowly (105 g/d, P < 0.05) and consumed less feed (280 g/d, P < 0.05) but had a similar (P > 0.05) gain:feed ratio to pigs reared in the unrestricted floor space. Pigs reared in the unrestricted floor space had greater (P < 0.05) predicted protein and lipid accretion rates throughout the growth period than pigs reared in the restricted floor space. Differences between genders for growth traits and carcass measurements were in agreement with previous research. The differences in growth performance, carcass measures, and compositional growth curves between these two sire lines were similar in the two floor spaces. PMID- 12772837 TI - Effect of transportation and commingling on the acute-phase protein response, growth, and feed intake of newly weaned beef calves. AB - The objective of this study was to investigate the effect of transportation and commingling on measures of the acute-phase protein response in newly weaned beef calves. Thirty-two (Exp. 1; average BW = 266 +/- 20.8 kg) and thirty-six (Exp. 2; average BW = 222 +/- 34.6 kg) Brahman-crossbred calves were randomly allotted to one of four treatments (2 x 2 factorial arrangement [transportation x commingling] in a completely randomized design). Body weight and jugular blood were collected at weaning, after shipment, and 1, 3, and 7 d after transport for Exp. 1, and at weaning and 1, 5, 9, 13, 17, and 21 d after transport for Exp. 2. Feed intake within pen was recorded daily for Exp. 2. Plasma fibrinogen, ceruloplasmin, haptoglobin, and cortisol concentrations were determined for all collection times. Additionally, serum amyloid-A and alpha-acid glycoprotein concentrations were determined in Exp. 1 and 2, respectively. In Exp. 2, commingled calves tended (P = 0.13) to have a higher DMI than noncommingled calves (5.3 and 4.8 kg/d, respectively). Transported calves lost more BW than nontransported calves from the time of weaning to d 1 (2.0 and 3.1% more BW loss for Exp. 1 and 2, respectively). With the exception of haptoglobin in Exp. 1, each of the acute-phase proteins measured in these studies increased over each sampling day. In Exp. 1, transported calves had higher (P < 0.05) mean serum amyloid-A concentrations than nontransported calves (48.9 vs. 33.4 microg/mL). There was a significant sampling day x transportation interaction (P < 0.01) for fibrinogen, ceruloplasmin, and haptoglobin in Exp. 1; transported calves had higher concentrations of fibrinogen following transport and on d 2 and 3, and ceruloplasmin on d 3. Haptoglobin concentrations were higher (P = 0.04) in nontransported calves on d 1 and 2 of Exp. 1. In Exp. 2, overall mean haptoglobin concentrations were higher in nontransported vs. transported calves. The results of these studies indicate that stressors associated with transportation affect the acute-phase protein response in newly weaned beef calves. More research is needed to determine whether these proteins might be valuable indicators of stress following the weaning process. PMID- 12772839 TI - Effect of early weaning on the performance of three-year-old, first-calf beef heifers and calves reared in the subtropics. AB - The objective of this study was to investigate the effectiveness of early weaning fall-born calves on heifer and calf performance in Florida. Over two consecutive years, 3-yr-old Braford and Brahman x Angus first-calf heifers were assigned randomly to one of two treatments; early-weaned (EW, n = 20 and 30 for yr 1 and 2, respectively) and normal-weaned (NW, n = 20 and 38 for yr 1 and 2, respectively). Calves were EW on January 23 and 3 for yr 1 and 2, respectively. Following EW, all first-calf heifers were returned to bahiagrass (Paspalum notatum) pastures with the mature cowherd. Early-weaned calves were maintained on annual ryegrass (Lolium multiflorum) pastures at 8.2 and 10.7 calves/ha for yr 1 and 2, respectively, and were provided supplemental grain mixture (14% CP) at 1.0% of BW daily. Normal-weaned calves remained with their dams in the mature cowherd on bahiagrass. Final calf BW was collected on April 17 (d 84) and April 24 (d 111) for yr 1 and 2, respectively. Early-weaned calves had a greater (P < 0.001) ADG (0.17 kg/d) in yr 1, but a lower (P < 0.001) ADG (-0.24 kg/d) in yr 2 compared with NW calves. Early weaning resulted in heavier first-calf heifers with greater BCS at the time of normal weaning (August 1; 491 vs. 452 kg, with BCS = 6.34 vs. 4.75 for EW and NW heifers, respectively; SEM = 5.0 and 0.07). Heifers with EW calves had a higher (P < 0.07) pregnancy rate during both years than normal-weaned heifers (89.5 vs. 50.0 and 96.7 vs. 80.0% pregnant during yr 1 and 2, respectively). Early-weaned, first-calf heifers also had a lower (P < 0.05) calving interval in yr 2 (384 vs. 404 d; SEM = 6.0). These data suggest that EW will improve body condition of first-calf heifers resulting in an increased pregnancy rate. Early-weaned calves maintained on winter ryegrass provide producers with the ability to optimize early-weaned calf performance, while capitalizing on low cost of gain and favorable spring markets. PMID- 12772840 TI - Growth performance, dry matter and nitrogen digestibilities, serum profile, and carcass and meat quality of pigs with distinct genotypes. AB - We investigated the effect of distinct genotypes on growth performance, DM and N digestibilities, serum metabolite and hormonal profiles, and carcass and meat quality of pigs. Eight control-line and eight select-line pigs with an equal number of gilts and castrated males per genotype were chosen from the group of pigs subjected to selection for lean growth efficiency. Pigs were housed individually and allowed ad libitum access to common grower, finisher 1, and finisher 2 diets when they reached approximately 20, 50, and 80 kg, respectively, and water throughout the study. Although genotype had no effect on growth performance during the finisher 2 phase and overall, select-line pigs grew faster and more efficiently (P < 0.05) during the grower and finisher 1 phases than did control-line pigs. Dry matter and N digestibilities during the grower phase were lower (P < 0.05) in select-line pigs compared with control-line pigs. Select-line pigs had less ultrasound backfat (P < 0.05) at the end of the grower and finisher 2 phases. Serum urea N (P < 0.05) and leptin concentrations were lower in select line pigs than in control-line pigs, but the effect of genotype on serum glucose, triglyceride, or insulin concentration was rather inconsistent. Select-line pigs had heavier heart (P < 0.05), liver (P = 0.08), and kidneys (P < 0.01), implying a higher metabolic activity. Less 10th-rib carcass backfat (P < 0.01) and a trend for larger carcass longissimus muscle area (P = 0.10) were reflected in the greater (P < 0.01) rate and efficiency of lean accretion in select-line pigs. Select-line pigs had lower subjective meat color (P < 0.01), marbling (P < 0.05), and firmness (P < 0.01) scores. Final serum leptin concentration was correlated positively with carcass backfat thickness (r = 0.73; P < 0.01) and negatively with overall feed intake (r = -0.77; P < 0.01). These results indicate that pigs with distinct genotypes exhibited differences in the growth rate, metabolite and hormonal profiles, and body composition. Further research is necessary to determine whether pigs with distinct genotypes respond differently to dietary manipulations, which would have an effect on developing optimal feeding strategies for efficient and sustainable pig production. PMID- 12772841 TI - Evaluation of strategies for selection for lean growth rate in pigs. AB - Lean growth rate (LGR) in pigs is a nonlinear biological function of growth rate and lean quantity. According to animal breeding theory, genetic progress for LGR is maximized with selection on a linear index of its component traits, but selection on direct EBV for LGR is also common. In this study, the performance of five criteria for selection on estimated LGR in pigs was evaluated through simulation over five generations: linear indexes of multiple-trait EBV of component traits with or without updating index weights in each generation; a nonlinear index of multiple-trait EBV of component traits; and direct selection on EBV for LGR from a single-trait model or a multiple-trait model that included LGR and component traits. The nonlinear index yielded the highest response in LGR in Generation 5, but the linear index with updating performed almost as well. Not updating weights for the linear index reduced response in LGR by 1.1% in Generation 5 (P < 0.05). Direct selection on single-trait EBV for LGR yielded the lowest responses in Generation 5. Direct selection on EBV for LGR from a multiple trait animal model yielded a 3.1% greater response in LGR in Generation 5 than direct selection on EBV for LGR based on a single-trait animal model (P < 0.05), but yielded a 1.9% lower response than the nonlinear index. Although differences in response in LGR were limited, alternative selection criteria resulted in substantially different responses in component traits. Linear index selection for LGR placed more emphasis on lean quantity, whereas direct selection for LGR emphasized growth rate. Based on the relative changes in the responses in LGR, selection for estimated LGR based on a nonlinear index or a linear index with updating is recommended for use in the swine industry. PMID- 12772842 TI - Quantitative trait locus detection in commercial broiler lines using candidate regions. AB - A QTL that explained a large proportion of the phenotypic difference between broiler and layer chickens in an experimental cross was evaluated in a commercial broiler line. A three-generation design, consisting of 15 grandsires, 608 half sib hens, and more than 50,000 third-generation offspring, was implemented within the existing breeding scheme of a broiler breeding company. Four markers from a candidate region on chicken chromosome 4 were selected for their informativeness in the grandsires and used to genotype the first two generations. Using half-sib analyses, linkage was studied between these markers and 13 growth and carcass traits. The QTL analyses confirmed the presence of significant QTL for body weight (P < 0.01) and residual feed intake (P < 0.05) on chicken chromosome 4. Furthermore, evidence was found for QTL affecting the relative weight of bone and muscle in the thigh. Four more markers were added to increase resolution of the QTL positions. This increased the significance of the QTL for body weight (P < 0.001) and residual feed intake (P < 0.01) and showed evidence (P < 0.05) for additional QTL affecting carcass weight and conformation score. This study showed for the first time that a QTL that explains differences between broilers and layers was segregating in lines that have been selected for body weight over 50 generations. A possible explanation could be a pleiotropic or closely linked effect on fitness-related traits that are not part of the present study. The results demonstrate the feasibility of QTL detection and the potential for marker assisted selection within a commercial broiler line without altering the existing breeding scheme. PMID- 12772844 TI - Comparison of different methods to assess the composition of pig bellies in progeny testing. AB - The objective of the study was to validate methods that assess the belly composition of stationary tested progenies of Pietrain boars. In German performance test stations, there are currently three methods of determining belly compositon in use: 1) a regression equation that contains different carcass characteristics, such as fat thickness and muscle area; 2) planimetric analysis of video or digital images acquired at the cut between the 13th and 14th ribs; and 3) estimation of the belly composition using ultrasound data from a three dimensional ultrasound image produced an online carcass-grading system. Validation of these techniques was performed on 400 carcasses of stationary tested Pietrain and Pietrain-sired crossbred pigs, which were slaughtered at a mean carcass weight of 85 and 97 kg. Magnetic resonance imaging (MRI) served as a reference to determine the lean content of the bellies. The correlation to MRI lean content ranged from 0.71 to 0.81, and corresponding correlation values were 0.62 to 0.64 for the digital imaging technique, and 0.53 to 0.59 for the AutoFOM online carcass-grading system. An increase in precision was achieved when information from digital imaging and linear carcass measures were included in the regression equation. Accuracy of the AutoFOM system does not seem to be sufficient to assess the belly composition for the special breeds in performance testing. However, extracting and combining 127 AutoFOM-base recordings into modified equations using partial least squares techniques yielded an improvement in the prediction accuracy for all tested breed and/or weight groups. PMID- 12772843 TI - The effects of thermal environment and spray-dried plasma on the acute-phase response of pigs challenged with lipopolysaccharide. AB - Forty barrows (TR4 x C22) were weaned at 17 d of age (BW = 6.27 +/- 0.30 kg), housed (two pigs/pen) in a thermal-neutral environment (TN; constant 26.7 degrees C), and fed diets with or without 7% (as-fed basis) spray-dried plasma (SDP). On d 7, one pig/ pen was moved into a cold environment (CE; constant 15.6 degrees C). Pigs were fitted with jugular catheters on d 11. On d 12, 16 pigs per environment (eight pigs per dietary treatment) were challenged i.v. with 75 microg of lipopolysaccharide (LPS)/kg of BW. Blood samples were collected over a 4.5-h period. Pigs were then killed and tissue samples were harvested for messenger RNA (mRNA) analysis. From d 0 to 7, pigs fed SDP diets had a lower gain:feed ratio (G/F) than pigs fed no SDP (533 +/- 14 vs. 585 +/- 17 g/kg; P < 0.03). Pigs housed in the CE consumed more feed and had a lower G/F than pigs housed in TN from d 7 to 11 (P < 0.001). There were no environment x diet interactions from d 7 to 11 (P > 0.78). Baseline concentrations of serum ACTH and cortisol were lower in the TN pigs than in the CE pigs (P < 0.001). Pigs fed diets without SDP had lower serum cortisol concentrations over the 4.5-h period than pigs fed SDP (time x diet, P < 0.001). Serum concentrations of tumor necrosis factor-alpha (TNF-alpha) were highest for pigs consuming SDP in the CE, whereas there were no differences among the other treatments (time x diet x environment, P < 0.02). Pigs housed in the CE had higher serum interleukin-1beta (IL-1beta) (P < 0.001) and interleukin-6 (IL-6; P < 0.001) than TN pigs. Pigs fed SDP also had slightly higher serum IL-1beta concentrations (P < 0.10) and higher (P < 0.001) IL-6 concentrations than pigs fed no SDP. Pigs fed SDP had 9% lower liver and 13% lower thymus mRNA expression of tumor necrosis factor-alpha (TNF alpha) than pigs that consumed no SDP (P < 0.06). Liver IL-1beta, IL-6, and LPS binding protein mRNA were higher in the CE than in the TN (P < 0.03, P < 0.001, and P < 0.05; respectively). In addition, spleen TNF-alpha (P < 0.03) and IL-6 (P < 0.01) mRNA levels were higher in the CE than in the TN. Pigs consuming SDP and challenged with LPS responded with elevated serum concentrations of cortisol and cytokines compared with pigs fed diets with no SDP. Housing pigs in a CE increased the baseline concentrations of ACTH and cortisol, and when coupled with an LPS challenge, resulted in elevated serum and tissue mRNA levels of cytokines. Cold stress and feeding SDP during a LPS challenge may result in increased stress and immune responses in young pigs. PMID- 12772845 TI - Amino acid sequences of multiple fast and slow troponin T isoforms expressed in adult bovine skeletal muscles. AB - Multiple nucleotide sequences of complementary DNA (cDNA) of bovine troponin T (TnT) isoforms expressed in the adult skeletal muscles were determined to facilitate the elucidation of the TnT degradation progress during postmortem aging of muscles. Fresh muscle samples were excised from the lingual, masseter, pectoralis, diaphragm, psoas major, longissimus thoracis, spinnalis, semitendinosus, semimembranosus, and biceps femoris muscles of three Holstein cows within 1 h of slaughter. Complementary DNA fragments of fast and slow TnT isoforms expressed in each muscle were amplified by reverse-transcribed PCR. Consequently, four major fragments of fast TnT and two fragments of slow TnT, all of which contained the complete coding region, were obtained. The sequence determination of these fragments revealed that at least eight and two isoforms were generated by the alternative splicing from bovine fast and slow TnT messenger RNA, respectively. In the fast TnT isoforms, five small variable exons were observed; three of these five exons were in the amino (N)-terminal region. The calculated molecular weight of fast and slow TnT isoforms ranged from 29,816 to 32,125 and from 30,166 to 31,284, respectively. The deduced amino acid sequences revealed that the N-terminal region of all the TnT isoforms was extremely glutamic acid-rich. Reverse-transcribed PCR analysis revealed that expression of each of these isoforms was distributed in a fast or slow muscle specific manner. Given that TnT degradation has been reported to accompany a decrease in glutamic acid content in the conventional 30-kDa degradation product, the sequence data suggested that the 30-kDa fragment seem to be generated by the proteolytic removal of the glutamic acid-rich N-terminal ends. The multiplicity of TnT isoforms may result in a complicated pattern of TnT degradation on SDS PAGE gel during beef aging. PMID- 12772846 TI - Evaluation of the E+V video image analysis system as a predictor of pork carcass meat yield. AB - This study was conducted to assess the ability of the VCS2001 (E+V, Oranienburg, Germany) video image analysis system to predict pork carcass composition. Pork carcasses (n = 278) were selected from a commercial packing plant to differ in weight, Fat-O-Meater (FOM) predicted percentage lean, and gender. Carcasses were imaged three times with the VCS2001, chilled overnight, and then sequentially fabricated into boneless subprimals. The VCS2001 accurately predicted the weight of total saleable product (R2 = 0.88, root mean square error [RMSE] = 1.84) and fat-corrected lean (R2 = 0.92, RMSE = 1.66), but autocorrelation existed between dependent and independent variables. The VCS2001 was acceptably accurate and precise in predicting weights of bone-in ham (R2 = 0.83, RMSE = 0.80), bone-in loin (R2 = 0.74, RMSE = 1.17), loin lean (R2 = 0.77, RMSE = 0.82), belly (R2 = 0.78, RMSE = 0.94), sparerib (R2 = 0.55, RMSE = 0.28), and boneless shoulder (R2 = 0.73, RMSE = 0.79). Weights were more accurately predicted than yields (as a percentage of hot carcass weight) of total saleable product (R2 = 0.47, RMSE = 1.97) or total fat-corrected lean (R2 = 0.44, RMSE = 1.89) using VCS2002, and it did not accurately predict percentages of bone-in ham (R2 = 0.45, RMSE = 1.13), ham lean (R2 = 0.32, RMSE = 1.46), bone-in loin (R2 = 0.29, RMSE = 1.36), loin lean (R2 = 0.56, RMSE = 0.90), belly (R2 = 0.43, RMSE = 1.08), sparerib (R2 = 0.08, RMSE = 0.32), or boneless shoulder (R2 = 0.30, RMSE = 0.88). New prediction models and equations were developed using VCS2001 output variables plus hot carcass weight to predict weight of total saleable product (R2 = 0.89, RMSE = 1.72) and fat-corrected lean (R2 = 0.93, RMSE = 1.55) with very minimal increases in accuracy and precision over that achieved using E+V-programmed models and equations. Use of new prediction models and equations marginally improved accuracy and precision of estimations of total saleable product yield (R2 = 0.56, RMSE = 1.81) and fat-corrected lean yield (R2 = 0.57, RMSE = 1.67) over that achieved using E+V-programmed models and equations. The VCS2001 was not able to predict pork carcass composition more accurately than existing technology; therefore, further development is needed to assure commercial viability of this instrument. PMID- 12772847 TI - Dietary betaine supplementation affects energy metabolism of pigs. AB - The effect of dietary betaine supplementation on energy partitioning in growing pigs under energy-restricted dietary conditions was assessed. The effect of betaine on the adaptation in energy metabolism of pigs over time after a change in diet and housing also was studied. Six groups of 14 group-housed barrows were assigned to one of two experimental diets: control or betaine-supplemented (0 or 1.29 g/kg of feed). Diets were corn- and soybean meal-based and were formulated to be limiting in energy content but sufficient in amino acids. The experiment comprised a 3-wk adaptation and a 3-wk experimental period. At the start of the experimental period, initial BW was 46 kg, each group of pigs was housed in a climate-controlled respiration chamber, and all pigs were subjected to a change in diet. During the experimental period, diets were diluted with 10% oat hulls. Pigs were fed at 2.5 times the energy requirements for maintenance, and during the experimental period, heat production, energy, and nitrogen balances were measured weekly. Metabolizibility of energy did not differ (P > 0.10) between diets. Averaged over the experimental period, betaine reduced heat production (P < 0.05) and energy requirements for maintenance (P < 0.10) and consequently increased energy retention (P < 0.10). Moreover, the difference in heat production between diets increased with time (P < 0.05). Similarly, the effect of betaine on the energy requirements for maintenance changed with time (P < 0.05). Maintenance requirements were similar in wk 1 and were decreased by betaine supplementation by 5.5% during wk 3 (477 vs. 452 kJ/[kg(0.75) x d]). Results of this study show that dietary betaine supplementation affects energy partitioning by growing pigs. However, based on the observed time-related changes in energy partitioning, it was concluded that dietary betaine supplementation did not influence adaptation by pigs to a change in housing and diet. PMID- 12772848 TI - Effects of two different dietary fermentable carbohydrates on activity and heat production in group-housed growing pigs. AB - The effects of two sources of dietary fiber (DF) on behavior and heat production (HP) in group-housed growing pigs were studied. Twenty clusters of 14 barrows (50 kg) were fed one of 10 diets. Diets differed mainly in type and content of fermentable DF (fDF) and in content of digestible starch. Five diets contained solvent-extracted coconut meal (SECM) and five diets contained soybean hulls (SBH) as the main fDF source. On an as-fed basis, pigs received 3.5, 13.2, 23.0, 32.7, or 42.4 g x kg(-0.75) x d(-1) of SECM or SBH. A total of 280 crossbred growing pigs were used, divided into clusters of 14 pigs each. Pigs were group housed and fed at 2.5 times the assumed maintenance energy requirements. All clusters were fed similar amounts of NE, ileal-digestible protein and amino acids, vitamins, and minerals. Consequently, DMI differed among diets because NE content decreased with increasing DF content. After a 32-d preliminary period, HP was measured per cluster during a 7-d experimental period in environmentally controlled respiration chambers. Behavior of the pigs was recorded using time lapse video recordings during two different days within the experimental period. Intake of digestible starch and fDF was different (P < 0.001) among diets, whereas intake of digestible CP was similar among diets. On average, pigs spent 153 min standing, 42 min sitting, 202 min lying on their chest, and 1,043 min lying on their flanks each day. Pigs fed SECM diets spent, on average, less time (P < 0.05) lying on their chest than pigs fed SBH diets. Total time spent on physical activity (i.e., standing plus sitting, 195 min/d) was not affected by diet. Total HP and resting HP were affected by diet and were on average lower (P < 0.01) for pigs fed SECM diets than for pigs fed SBH diets. Activity-related heat production (AHP) averaged 65 kJ x kg(-0.75) x d(-1) and was not affected by diet. There was a linear relationship (P < 0.001) between fDF intake and HP, but there was no relationship between fDF intake and AHP. During different parts of the day, fDF intake also affected HP. The saving effect of physical activity on the NE values of fDF from SECM and SBH were 0.56 and 0.84 kJ/g of fDF intake, respectively. Neither of these saving effects was significantly different from zero. We conclude that fDF from SECM and SBH did not affect energy expended on physical activity by growing pigs, and that the NE value of fDF from SECM and SBH was not affected by changes in physical activity. PMID- 12772849 TI - Effect of fishmeal replacement with spray-dried animal plasma and colistin on intestinal structure, intestinal microbiology, and performance of weanling pigs challenged with Escherichia coli K99. AB - We evaluated spray-dried animal plasma (SDAP) as an alternative to antimicrobial medication with colistin sulfate in weanling pigs challenged with Escherichia coli K99. Forty-eight piglets weaned at 24 d of age were distributed into 12 pens, and each pen was assigned to one of four dietary treatments. All the piglets were given an oral dose of 5 x 10(7) cfu of E. coli K99 at weaning. The dietary treatments followed a factorial arrangement with two levels of SDAP (0 and 7%) and two levels of colistin (0 and 300 mg/kg of diet). The ADG and ADFI were measured on d 7 and 14 of trial. Three piglets from each treatment were killed on d 7 and 14 to remove the small intestine, and to obtain ileal and cecal digestive contents. The inclusion of SDAP improved ADG by 68 g (P < 0.05) and ADFI by 41 g (P < 0.10) in wk 1 of trial. During wk 2, SDAP improved ADG by 41 g (P < 0.10) and gain:feed ratio (G:F) by 25% (P < 0.01). On the other hand, whereas colistin had no effect on performance in wk 1, it improved ADG by 102 g (P < 0.01), ADFI by 62 g (P < 0.01), and G:F by 26% (P < 0.01) in wk 2. Over the 14 d of the trial, ADG was improved by 54 (P < 0.05) and 75 g (P < 0.05), and G:F was improved by 35 (P < 0.05) and 32% (P < 0.05) due to SDAP and colistin, respectively. There was interaction between colistin and SDAP for ADFI in wk 2 and between d 0 to 14 (P < 0.05), which indicates that their effects were not additive. The use of colistin was advantageous in the maintenance of the integrity of the intestinal mucosa of the pigs, as suggested by a small intestine that was 93 g heavier (P < 0.10) and with the tallest villi 106 microm longer (P < 0.10) than in pigs without colistin. The inclusion of SDAP in the diet favored the growth of lactobacilli in the ileum (P < 0.10) and the cecum (P < 0.05), whereas colistin reduced the number of enterococci in the cecum (P < 0.05) and of Escherichia coli both in the ileum and the cecum (P < 0.001). These results suggest that SDAP may be an alternative to medicated feed with antibiotics since it provided a level of protection against an experimental challenge with E. coli K99 similar to that obtained with colistin, an antibiotic of proven efficacy. The current situation in which the use of antimicrobials in animal feeding is being questioned should encourage further investigation into the use of SDAP as a means of preventing disease in pigs at weaning. PMID- 12772850 TI - Effect of dietary energy concentration and apparent ileal digestible lysine:metabolizable energy ratio on nitrogen balance and growth performance of young pigs. AB - Two experiments were conducted to determine the optimal apparent ileal digestible lysine:ME (Lys:ME) ratio and the effects of lysine and ME levels on N balance (Exp. 1) and growth performance (Exp. 2) in growing pigs. Diets were designed to contain Lys:ME ratios of 0.6, 0.7, 0.8, and 0.9 g/MJ at 13.5 and 14.5 MJ of ME/kg of diet in a 4 x 2 factorial arrangement. In Exp. 1, conventional N balances were determined on 48 crossbred barrows (synthetic line 990, initial BW = 13.1 +/- 0.7 kg) at approximately 15, 20, and 25 kg of BW with six pigs per diet. At 15 kg of BW, an energy density x Lys:ME ratio interaction on daily N retention was observed (P < 0.05). At each BW, N retention improved with an increase in N intake associated with increasing ME concentration. In 15-kg BW pigs, increasing the Lys:ME ratio increased daily N retention at the 13.5 (linear, P < 0.001) and 14.5 MJ of ME level (linear, P < 0.01; quadratic, P < 0.05). In 20-kg BW pigs, N retention (g/d) increased (linear, P < 0.001; quadratic, P < 0.01) and N retention (percentage) increased (linear, P < 0.001) as the Lys:ME ratio increased. At 25 kg of BW, N retention (g/d) increased quadratically (P < 0.05) with an increase in Lys:ME ratio. The Lys:ME ratios that maximized daily N retention at 15 kg of BW were 0.88 and 0.85 g/MJ at the 13.5 and 14.5 MJ of ME levels, respectively and 0.81 and 0.77 g/MJ (for both ME levels) at 20 and 25 kg of BW, respectively. Over the 28-d trial, an energy density x Lys:ME ratio interaction on ADG was observed (P < 0.05). Increasing energy density increased growth performance, whereas increasing the Lys:ME ratio in high-energy diets increased ADG (linear, P < 0.05; quadratic, P < 0.01) and gain:feed ratio (G/F) quadratically (P < 0.01). Average daily gain and G/F ratio were greatest in pigs fed the 14.5 MJ of ME diet and the Lys:ME ratio of 0.82 g/MJ. In Exp. 2, 128 individually housed crossbred barrows and gilts (initial BW = 12.8 +/- 1.6 kg) were used to determine the effect of diets used in Exp. 1 on growth performance in a 4 x 2 x 2 factorial arrangement. The ME level increased ADG and G/F from d 0 to 14 and from d 0 to 28. Increasing the Lys:ME ratio increased ADG from d 0 to 14, whereas growth performance was maximized in pigs fed Lys:ME ratio of 0.82 g/MJ. These results suggest that pigs from 13 to 20 and from 20 to 30 kg of BW fed diets containing 14.5 MJ of ME/kg had maximum N retention and ADG at 0.85 and 0.77 g of apparent ileal digestible lysine/MJ of ME, respectively. PMID- 12772851 TI - The toxicity and kinetics of larkspur alkaloid, methyllycaconitine, in mice. AB - Larkspur poisoning sporadically kills from 5 to 15% of the cattle on North American mountain rangelands. Of the 40 different diterpenoid larkspur alkaloids, the one that is thought to be responsible for much of the toxicity has been identified as methyllycaconitine (MLA). Little is known of MLA toxicokinetics or excretion. The purpose of this study was to further characterize the clinical effects of MLA toxicity in mice and determine the toxicokinetics of MLA excretion. Eight groups of mice were dosed intravenously with 2.0 mg/kg of BW of MLA, killed, and necropsied at 0, 1, 2, 5,10,15, 30, and 60 min after injection. Treated animals were reluctant to move, trembled, and developed dyspnea, muscular twitches, and convulsions. Within several minutes, the clinical signs abated and behavior slowly returned to normal over approximately 20 min. At necropsy serum, brain, liver, kidney, and skeletal muscle were collected and frozen. Blood and tissues were extracted and analyzed for MLA with HPLC and electron spray mass spectrometry. Blood MLA elimination followed a normal biphasic redistribution and excretion pattern (r = 0.99) with a K of elimination of 0.0376 and half-life of 18.4 min. Other tissues had similar clearance rates. These data indicate the MLA is rapidly distributed and excreted. In mice, the clinical effects of poisoning seem to affect the central nervous system, causing dyspnea and "explosive" muscular twitches and convulsions. Because livestock commonly eat larkspur at subclinical doses, they are likely to have larkspur alkaloids in many tissues. These results suggest that animals exposed to larkspur should rapidly excrete MLA (within several hours) and that the residues in animal tissues are not likely to be a problem if animals are given several days to allow toxin clearance. PMID- 12772853 TI - Effect of repeated ketoprofen administration during surgical castration of bulls on cortisol, immunological function, feed intake, growth, and behavior. AB - To determine the effect of repeated ketoprofen (K) administration to surgically castrated bulls on cortisol, acute-phase proteins, immune function, feed intake, growth and behavior, 50 Holstein x Friesian bulls (11 mo old; 300 +/- 3.3 kg) were assigned to one of five treatments: 1) untreated control (C); 2) surgical castration at 0 min (S); 3) S following an i.v. injection of 3 mg/kg of BW of K at -20 min (SK1); 4) S following 1.5 mg/kg of BW of K at -20 and 0 min (SK2); or 5) S following 1.5 mg/kg of BW of K at -20 and 0 min and 3 mg/kg of BW of K at 24 h (SK3). Castration acutely increased plasma cortisol concentrations in S- and K treated animals compared with C, with no differences in peak and interval to peak cortisol responses among the castration groups. Overall, the integrated cortisol response was greater (P < 0.05) in the castrates than in C, whereas K treatments decreased (P < 0.05) this response compared with S alone, with no differences between K treatments. Plasma haptoglobin and fibrinogen concentrations were increased (P < 0.05) on d 3 in the castration groups compared with C as the result of tissue trauma induced by castration, whereas SK1 and SK2 had lower (P < 0.05) haptoglobin concentrations than S animals. On d 1, concanavalin A-induced interferon-gamma production was suppressed (P < 0.05) in S and SK3 compared with C, SK1, and SK2 animals. Overall from d 1 to 33, DMI were lower (P < 0.05) in S, SK1, and SK3 than in C animals. From d -1 to 35, ADG were lower (P < 0.05) in S, SK2, and SK3 compared with C animals. A higher (P < 0.05) incidence of standing postures and lower incidence of lying postures was observed in S compared with C during the first 6 h after treatment. However, the higher (P = 0.02) incidence of abnormal standing activities observed for S was reversed (P < 0.05) by the K treatments. In conclusion, surgical castration increased plasma cortisol and acute-phase proteins and decreased immune function, feed intake, and growth rate. Ketoprofen effectively reduced the cortisol response to castration, but there was no advantage in treating with two split doses of K (1.5 mg/kg of BW per dose). A repeated K dose 24 h after treatment (3 mg/kg of BW) had no influence on changes in acute-phase proteins and immune response. Systemic analgesia with K is an effective method for alleviating acute inflammatory stress associated with castration. PMID- 12772852 TI - Progesterone increases susceptibility of gilts to uterine infections after intrauterine inoculation with infectious bacteria. AB - In cattle and sheep, a progestogenated uterus is susceptible to infections, but this is not well documented for pigs. Therefore, the effects of day of the estrous cycle and progesterone on the susceptibility to uterine infections were evaluated. Gilts (n = 5 per group) were assigned to treatments in 2 x 2 factorial arrays. In Exp. 1, day of cycle and bacterial challenge were main effects. On d 0 or 8, uteri were inoculated with either 70 x 10(7) cfu of Escherichia coli and 150 x 10(7) cfu of Arcanobacterium pyogenes in PBS or with PBS. In Exp. 2, ovariectomy (OVEX) and progesterone treatment were main effects. On d 0, gilts were ovariectomized or a sham procedure was performed. After surgery, gilts received i.m. injections of progesterone (10 mg/5 mL) or 5 mL of safflower oil diluent twice daily. On d 8, gilts were inoculated with the same doses of bacteria as in Exp. 1. In Exp. 1 and 2, vena caval blood was collected for 4 d, after which uteri were collected. Sediment and ability to culture E. coli and A. pyogenes from uterine flushings were used to diagnose infections. Differential white blood cell counts and lymphocyte response to concanavalin A (Con A) and lipopolysaccharides (LPS) were used to measure lymphocyte proliferation. Progesterone, estradiol-17beta, prostaglandin F2alpha, (PGF2alpha), and prostaglandin E2 (PGE2) were measured in vena caval blood. In Exp. 1, d-8 gilts receiving bacteria developed infections, but d-0 gilts receiving bacteria did not. Daily percentages of neutrophils and lymphocytes changed (P < 0.05) with cycle day and bacterial challenge. Basal- and Con A-stimulated lymphocyte proliferation were greater (P < 0.05) for d-0 than for d-8 gilts. Concentrations of PGF2, (P < 0.01) and PGE2 (P < 0.05) increased after bacterial challenge, regardless of stage of the estrous cycle at the time of inoculation. In Exp. 2, OVEX decreased (P < 0.001) and progesterone treatment increased (P < 0.001) progesterone concentrations, and OVEX decreased (P < 0.01) estradiol-17beta. Gilts with ovarian and/or exogenous progesterone developed infections. Daily percentages of neutrophils and lymphocytes changed in response to OVEX, and neutrophils changed (P < 0.05) in response to endogenous and exogenous progesterone. Lymphocyte proliferation in response to Con A and LPS increased (P < 0.05) with OVEX and decreased (P < 0.05) with progesterone treatment. We conclude that endogenous and exogenous progesterone reduce the ability of the uterus in gilts to resist infections. PMID- 12772854 TI - Relationships between uterine and fetal traits in rabbits selected on uterine capacity. AB - The aim of this study was to investigate whether uterine capacity (UC) in rabbits was related to uterine horn length and weight and whether these uterine traits and vascular supply were related to fetal development and survival. Data from 48 unilaterally ovariectomized (ULO) does of the High and 52 ULO does of the Low UC lines of a divergent selection experiment on UC were used. Does were slaughtered on d 25 of fifth gestation. The High line showed higher ovarian weight (0.08 g, P < 0.05) linked to a higher ovulation rate (1 ovum, P < 0.05) and greater length of the empty uterine horn. There were no differences between lines in the remaining doe traits. The number of implanted embryos and live fetuses, fetal survival, and uterine weight and length were positively associated and explained most of the observed variation. Average weights of the live fetuses and their fetal and maternal placentae were not related to uterine weight and length. The linear regression coefficient of full uterine horn length on the number of live fetuses was 2.43 +/- 0.21. The weight of the full uterine horn showed a small quadratic relationship (P < 0.05) with the number of live fetuses. Full uterine horn length, after adjusting for the number of embryos, was negatively associated (P < 0.001) with the number of dead fetuses. The linear regression coefficient of average fetal placental weight of the live fetuses on number of implanted embryos was higher (P < 0.10) in the Low line (-0.23 +/- 0.04 vs. -0.12 +/- 0.04). The linear regression coefficient of average weight of the live fetuses on the average weight of their fetal placentae was higher (P < 0.10) in the High line (2.56 +/- 0.47 vs. 1.27 +/- 0.57). The High line was more efficient, most likely because an increase in intrauterine crowding has a lesser effect on the development of fetal placentae and fetuses. The fetal position within the uterus did not affect the proportion of dead embryos. Fetuses with placentae receiving a single blood vessel had a higher probability of death (P < 0.001) and the lowest weight. There was no difference between lines for individual weight of the live fetuses, but the High line showed higher individual weights of fetal (P < 0.01) and maternal placentae (P < 0.10). Live fetuses in the midportion of the uterus were lighter in weight (P < 0.05) than in the oviductal and cervical regions (20.3 vs. 21.6 and 21.7g). Increasing uterine capacity increases uterine length and decreases weights of fetus and fetal placenta in rabbits. PMID- 12772855 TI - Central and peripheral concentrations of tumor necrosis factor-alpha in Chinese Meishan pigs stimulated with lipopolysaccharide. AB - The objective of this study was to investigate the presence of tumor necrosis factor-alpha (TNF-alpha) in the central nervous system and the effects of lipopolysaccharide on central and peripheral concentrations of TNF-alpha, behavioral conditions (standing or lying), elimination scores (defecation or urination), rectal temperature, and food intake (as-fed basis) in Chinese Meishan pigs. Intravenous injections of lipopolysaccharide resulted in increased (P < 0.05) plasma concentrations of TNF-alpha and cortisol. Although urination was not affected by the administration of lipopolysaccharide, defecation was stimulated (P < 0.05). Lipopolysaccharide increased (P < 0.05) rectal temperature and standing rate, and inhibited (P < 0.05) food intake in pigs. To determine whether TNF-alpha is present in the porcine central nervous system as well as in peripheral blood, TNF-alpha and its specific transcripts in brain tissues (hypothalamus, amygdala, or hippocampus) and the pituitary were determined. The abundance of TNF-alpha messenger RNA and immunoreactive TNF-alpha were observed in all tissues, and the concentrations of TNF-alpha were increased (P < 0.05) by the intramuscular injection of lipopolysaccharide. These results suggest that TNF alpha is present in the central nervous system, and plays some roles in its biological regulation in Chinese Meishan pigs. PMID- 12772856 TI - Effect of ketoprofen, lidocaine local anesthesia, and combined xylazine and lidocaine caudal epidural anesthesia during castration of beef cattle on stress responses, immunity, growth, and behavior. AB - To determine the effects of burdizzo castration alone or in combination with ketoprofen (K), local anesthesia (LA), or caudal epidural anesthesia (EPI) on plasma cortisol, acute-phase proteins, interferon-gamma production, growth, and behavior of beef cattle, 50 Holstein x Friesian bulls (13 mo old, 307 +/- 5.3 kg) were assigned to (n = 10/treatment): 1) control (handled; C); 2) burdizzo castration (B); 3) B following K (3 mg/ kg of BW i.v.; BK); 4) B following LA (8 mL into each testis and 3 mL s.c. along the line where the jaws of the burdizzo were applied with 2% lidocaine HCl; BLA); and 5) B following EPI (0.05 mg/kg of BW of xylazine HCl and 0.4 mg/kg of BW of lidocaine HCl as caudal epidural; BEPI). The area under the cortisol curve against time was lower (P < 0.05) in BK than in B, BLA, or BEPI animals. On d 1 after treatment, plasma haptoglobin concentrations were higher (P < 0.05) in B, BLA, and BEPI than in BK animals. On d 3, haptoglobin and plasma fibrinogen concentrations were higher (P < 0.05) in all castration groups than in C. On d 7, haptoglobin and fibrinogen concentrations remained higher (P < 0.05) in BLA than in B and C animals. On d 1, concanavalin A-induced interferon-gamma production was lower (P < 0.05) in B, BLA, and BEPI than in C, but there was no difference between BK and C animals. From d -1 to 35, ADG was lower (P < 0.05) in B, BLA, and BEPI animals, but not in BK compared with C animals. Overall, there was a higher (P < 0.05) incidence of combined abnormal postures in B than in C, BK and BEPI animals. Although the use of K and EPI decreased (P < 0.05) these postures compared with B alone or B with LA, there was no difference between the K and EPI treatment. In conclusion, burdizzo castration increased plasma cortisol and acute-phase proteins, and suppressed immune function and growth rates. Local anesthesia prolonged the increase in acute-phase proteins. Ketoprofen was more effective than LA or EPI in decreasing cortisol and partially reversed the reduction in ADG following castration. The use of K or EPI was more effective than LA in decreasing pain associated behavioral responses observed during the first 6 h after treatment. Systemic analgesia with ketoprofen, a non-steroidal antiinflammatory drug, was more effective in reducing inflammatory responses associated with castration than LA or EPI. PMID- 12772857 TI - Technical note: time-resolved immunofluorometric assay for growth hormone in ruminants. AB - A noncompetitive, time-resolved immunofluorometric assay (TRIFMA) was developed using a selected pair of monoclonal antibodies (mab) raised against recombinant bovine GH, with the catching mab immobilized on microtiter plate wells and the detection mab labeled with Eu3+ as a tracer, arranged as a sandwich. Plates were coated with mab1.15 (680 ng/well) using a phosphate buffer (pH 4.9), and then blocked with assay buffer containing 1% (wt/vol) BSA. The assay procedure involved incubation of 50 microL of sample (plasma or serum) and 200 microL of assay buffer containing 25 ng of mab1.2-Eu3+ conjugate for 4 h at 25 degrees C. Plates were then washed six times, incubated for 5 to 10 min with 250 microL of enhancement solution, and fluorescence read with a time-resolved fluorometer. The sensitivity of the assay was 0.1 ng/mL, and the working range was 0.2 to 200 ng/mL. Recovery of quantitative amounts of bovine GH added to plasma samples was close to 100%. Cross-reactivity with other bovine pituitary hormones or with GH from nonbovidae or cervidae species was not significant. Intra- and interassay CV during routine operation was 4.4 and 10.7%, respectively (mean = 3.54 ng/mL). Plasma concentrations of bovine GH determined by TRIFMA correlated closely (r2 > or = 0.93) with RIA results, with a conversion ratio of 0.62 when the higher specificity of the monoclonal antibodies was taken into account. The TRIFMA is a reliable alternative to RIA methods because the assay employs no radiolabeled or hazardous chemicals, delivers results rapidly, and has little risk of down periods. PMID- 12772858 TI - Effects of deslorelin acetate implants in horses: single implants in stallions and steroid-treated geldings and multiple implants in mares. AB - Three experiments were performed to test the following hypotheses: 1) stallions and/or progesterone-estradiol-treated geldings could serve as models for the effects of a single implant of the GnRH analog, deslorelin acetate, on LH and FSH secretion by mares; and 2) multiple implants of deslorelin acetate could be used as a means of inducing ovarian atrophy in mares for future study of the mechanisms involved in the atrophy observed in some mares after a single implant. In Exp. 1, nine light horse stallions received either a single deslorelin implant (n = 5) or a sham injection (n = 4) on d 0. In Exp. 2, 12 geldings received daily injections of progesterone on d -20 through -4, followed by twice-daily injections of estradiol on d -2 to 0. On the morning of d 0, geldings received either a single deslorelin implant (n = 6) or a sham injection (n = 6). Daily injections of progesterone were resumed on d 2 through 15. In Exp. 1, plasma LH and FSH were elevated (P < 0.05) in the treatment group relative to controls at 4, 8, and 12 h after implant insertion. In the treated stallions, FSH was decreased (P < 0.05) on d 3 to 13, and LH was decreased on d 6 to 13. In Exp. 2, plasma LH and FSH were elevated (P < 0.05) at 4,8, and 12 h after deslorelin implant insertion. Plasma LH was suppressed (P < 0.05) below controls on d 2 to 7, 9, and 11 to 15; plasma FSH was suppressed (P < 0.05) on d 4 to 15. In Exp. 3, 21 mares were used to determine whether multiple doses of deslorelin would cause ovarian atrophy. Mares received one of three treatments: 1) sham injections; 2) three implants on the first day; or 3) one implant per day for 3 d (n = 7 per group). Treatment with multiple implants increased (P < 0.05) the interovulatory interval by 14.8 d and suppressed (P < 0.01) LH and FSH concentrations for approximately 25 d; no mare exhibited ovarian atrophy. In conclusion, after an initial short-term increase in LH and FSH secretion, deslorelin implants caused long-term suppression of both gonadotropins in stallions as well as in geldings treated with progesterone and estradiol to mimic the estrous cycle. It is likely that either of these models may be useful for further study of this suppression in horses. Although multiple implants in mares suppressed gonadotropin secretion longer than a single implant, the lack of ovarian atrophy indicates that the atrophy observed after a single implant in previous experiments was likely due to the susceptibility of individual mares. PMID- 12772859 TI - Characterization of season and sampling method effects on measurement of forage quality in fescue-based pastures. AB - Information on seasonal changes and effects of sampling methods on the measurement of forage quality is limited for fescue-based pastures. Eight continuously grazed, 0.76-ha, fescue-based pastures were used to compare forage type, method of collection, and seasonal effects on forage quality in a repeated measures, split-plot design. Four pastures were interseeded with red clover in March 2000. Masticate (M; from four ruminally cannulated steers) and hand- clipped (C) samples were collected every 28 d from April to October 2000. Interseeding red clover did not affect (P > 0.10) OM, CP, NDF, and ADF concentrations or CP degradability. Sampling method and season interacted (P < 0.03) for OM, CP, NDF, and ADF concentrations. Concentrations of OM averaged 5 percentage units more (P < 0.01) in C than in M in all months and were more variable with M than with C. Samples clipped between April and September averaged 5.5 percentage units greater NDF (P < 0.01), 3.0 percentage units greater ADF (P < 0.01), and 4.5 percentage units less CP (P < 0.01) than masticate samples obtained during the same time period. Fiber and CP concentrations did not differ (P > 0.10) between C and M samples obtained in October. Differences in CP degradability estimates (using Streptomyces griseus protease) between the two sample types were greater in late-season samples than in samples obtained from April to June. When S. griseus protein degradability estimates were compared with in situ estimates for masticate samples, no differences (P > 0.10) were detected early in the season (April to June). However, the S. griseus procedure overestimated in situ values (P < 0.01) by an average of 3 percentage units in samples obtained between July and October. Differences in composition of C and M samples were substantial until late season, when opportunities for selective grazing were minimal. Small differences between S. griseus and in situ estimates of CP degradability indicate that the S. griseus procedure can yield useful CP degradability estimates for fescue-based pasture samples. Although it might be possible to apply correction values to clipped samples to estimate CP and fiber concentrations of diets selected by grazing cattle, inconsistent relationships preclude this approach for estimates of CP degradability. PMID- 12772860 TI - Use of nonergot alkaloid-producing endophytes for alleviating tall fescue toxicosis in sheep. AB - Nonergot alkaloid-producing endophytes from New Zealand were inserted into tall fescue (Festuca arundinacea) cultivars in an attempt to address the problem of fescue toxicosis in grazing sheep. A 3-yr grazing study was conducted to determine lamb performance and to evaluate toxicosis in lambs grazing nonergot alkaloid-producing endophyte-infected (AR542 or AR502), endophyte-free (E-), or wild-type toxic endophyte-infected (E+) Jesup tall fescue or nonergot alkaloid producing endophyte-infected (AR542) Georgia-5 tall fescue. Replicated 0.11-ha tall fescue paddocks were established at the central Georgia Branch Station during September 1997 and stocked with lambs from spring 1998 through autumn 2000. Mean ergot alkaloid concentrations were higher (P < 0.01) in E+ forage than in AR542, AR502, and E- tall fescue, and ergot alkaloid concentrations in E- plants and plants infected with AR542 and AR502 were low. Forage availability did not differ (P = 0.92) across treatments during autumn and was higher (P < 0.05) in Georgia-5 AR542 than in Jesup AR502 and E+ pastures. Initial serum prolactin (PRL) concentrations did not differ (P = 0.58) across treatments during autumn, but were higher on Jesup AR542 than E+ during spring. Post-treatment serum PRL concentrations were depressed (P < 0.01) on E+ compared with AR542, AR502, and E- in both spring and autumn. Signs of heat stress were observed in E+ lambs during periods of high ambient temperatures. Mean post-treatment rectal temperature and mean stocking rate exhibited treatment x year interactions (P < 0.05). Lamb ADG was higher (P < 0.05) on AR542, AR502, and E- than on E+ tall fescue. Similarly, gain/hectare was higher (P < 0.015) on AR542, AR502, and E- than on E+. Tall fescue pastures containing AR542 and AR502 endophytes yielded lamb performance that did not differ from that on E- tall fescue and which was superior to performance on E+ tall fescue. Depressed PRL concentrations and elevated rectal temperatures as indicators of toxicosis were evident only in lambs grazing E+ tall fescue, suggesting that nonergot alkaloid-producing endophyte-infected tall fescue is a viable alternative for alleviating tall fescue toxicosis. PMID- 12772861 TI - Influence of substrate and/or neurohormonal mimic on in vitro pancreatic enzyme release from calves postruminally infused with partially hydrolyzed starch and/or casein. AB - Our objectives were to determine the effects of neuroendocrine challenge and substrates on in vitro alpha-amylase and trypsin release in pancreatic tissue collected from Holstein calves (n = 24; 88 +/- 3 kg) abomasally infused for 10 d with tap water (control), partially hydrolyzed starch (SH; 4 g/[kg of BW x d]) and/ or casein (0.6 g/[kg of BW x d]). The caudal portion of the pancreas was removed, rinsed with ice-cold saline, cut into approximately 2 x 2-mm segments, and incubated in oxygenated Krebs Ringer bicarbonate buffer containing no substrate (control), glucose, amino acids, or VFA at 39 degrees C. After 60 min of incubation, neurohormonal mimics (none; control), carbachol (acetylcholine analog; 10 microM final), or caerulein (cholecystokinin mimic; 100 nM final) were added to the flasks and tissue was incubated for 60 min. Pancreatic tissue concentrations and in vitro release of alpha-amylase and trypsin decreased (P < 0.001) in calves abomasally infused with SH. Carbachol increased (P < 0.10) alpha amylase and trypsin release in tissue collected from all calves. An effect of caerulein to increase alpha-amylase release (P < 0.10) was only observed with prior exposure to abomasal casein infusion in vivo or with simultaneous incubation with amino acids in vitro. Caerulein increased (P < 0.10) trypsin release in tissue collected from all calves except for those receiving SH + casein. Glucose decreased (P < 0.10) alpha-amylase release from pancreatic tissue collected from calves receiving abomasal control and casein treatments. Amino acids decreased (P < 0.10) alpha-amylase and trypsin release from pancreatic tissue collected from calves receiving the abomasal control treatment. Glucose, amino acids, and VFA decreased (P < 0.10) trypsin release from tissue collected from calves receiving abomasal SH. These data indicate that carbachol can stimulate pancreatic enzyme release in vitro. Caerulein, however, is only effective in stimulating in vitro pancreatic enzyme release in tissue from calves with an increased postruminal protein supply or in tissue incubated with amino acids. The results indicate that postruminal and local nutrients might be important in altering the responsiveness to a neuroendocrine challenge and could be an important regulatory event involved with dietary adaptation in ruminants. PMID- 12772862 TI - Synchrony of nutrient supply to the rumen and dietary energy source and their effects on the growth and metabolism of lambs. AB - The objective of the current series of experiments was to assess the effects of dietary synchrony of OM and N supply to the rumen, achieved by altering the sequence of feeding individual ingredients and in diets with different energy sources, on the metabolism and performance of growing lambs. In Exp. 1, the in situ degradability coefficients of OM and N were determined for five feed ingredients and subsequently was used to formulate two diets, based either on barley or sugar beet pulp, to have a similar predicted nutrient content. Within each diet, specific ingredients were shifted between the 0900 and 1600 feeding to provide either a synchronous, intermediate, or asynchronous supply of OM and N to the rumen. In Exp. 2, these diets were fed at a restricted level to 48 growing lambs with an initial live weight of 25.1 +/- 4.22 kg and a slaughter weight of 41.4 +/- 1.94 kg. There was no significant effect of dietary treatment on live weight gain or feed conversion efficiency. Lambs fed the synchronous diets deposited more kidney knob and channel fat than lambs on the asynchronous or intermediate diets (P < 0.05), whereas lambs fed the barley-based diets deposited more carcass (P < 0.05) and noncarcass (P < 0.001) fat than lambs on the sugar beet-based diets. Lambs fed the asynchronous diets retained less energy over the course of the experiment than lambs on the intermediate or synchronous diets (P < 0.05), and had a lower energy efficiency (0.079, 0.097, and 0.093 MJ retained/ MJ of intake, respectively, P < 0.05). Lambs fed the barley-based diets retained more energy than lambs on the sugar beet-based (P < 0.001) and had a higher energy balance (0.095 vs. 0.084 MJ retained/MJ intake, respectively; P < 0.01). Plasma ammonia concentrations mirrored ruminal ammonia concentrations on the barley-based diets, but not sugar beet-based diets. In Exp. 3, lambs fed the sugar beet-based diets had a higher digestibility of OM and NDF (P < 0.001). By contrast, lambs on the barley-based diets had a higher level of purine derivative excretion and microbial N production (P < 0.001). The results indicate that neither dietary synchrony nor energy source significantly influenced growth rate. However, both the asynchronous and sugar beet pulp-based diets resulted in a lower efficiency of dietary energy use, and the avoidance of asynchronous patterns of nutrient release within the rumen can improve energy efficiency in growing lambs. PMID- 12772864 TI - Triage and approach to the acute abdomen. AB - The presentation of a patient with acute abdominal pain often is both a diagnostic and therapeutic challenge for the veterinary practitioner. Rapid physical examination and assessment of the patient's cardiovascular status are critical to initiating appropriate and often aggressive medical management as diagnostic tests are being performed. A number of diagnostic tests can be performed, including complete blood count, serum biochemistry profile, serum amylase and lipase, abdominal radiographs, abdominal ultrasound, abdominal paracentesis or diagnostic peritoneal lavage, cytology, and fluid analysis. In some cases, surgical exploration may be indicated, depending on each patient's clinical signs and response to therapy. Of the most important aspects of medical management of the patient with acute abdominal pain is to maintain oxygen delivery to tissues and to treat aggressively for pain or discomfort. The initial triage and approach to the patient with acute abdomen are discussed. PMID- 12772863 TI - A long-term study of knowledge, risk, and ethics for students enrolled in an introductory biotechnology course. AB - Since 1989, an undergraduate course addressing issues concerning biotechnology in agriculture has been taught annually. To determine the extent to which students were engaged in classroom instruction and assimilated knowledge from the course, students were asked to complete an "Attitudes About Biotechnology" questionnaire. Students were queried about their knowledge of biotechnology, perceptions of risks associated with biotechnology, and ethics and attitudes toward the use of biotechnology in agriculture. Responses to the survey questions were evaluated both pre-and postcourse. Data were collected from 13 classes over a 10-yr period. Inspection of the precourse grade point average and the course grades for these students revealed no trends up or down over the period of this study. Similarly, inspection of the data revealed no effect from problem-based learning, student facilitated group work, or technology on student attainment of knowledge, perception of risk, or ethical views. For each of the five knowledge questions, the average score increased as a result of taking the course. On two of the knowledge questions, the average score increase for females from pre- to postcourse was greater than for males. Based on our measurements using the postcourse questionnaire, the correlation between perceived and actual knowledge was not significantly different from zero. In two of the four risk questions, there was no change in average score as a result of the course; however, student perception of risk associated with genetic engineering of plants increased. Although average scores for student perception of risk due to the perceived impact of genetically engineered products on people or the environment decreased from pre- to postcourse assessment, the average score was higher (P < 0.01) for females than for males. Males were more accepting of genetic manipulation of cells in a laboratory than were females both pre- and postcourse. Although student knowledge of biotechnology was increased and the perceived risk due to biotechnology was altered, there was no evidence that students altered their ethical position on biotechnology as a result of this course. PMID- 12772865 TI - Radiographic techniques and interpretation of the acute abdomen. AB - Radiography is a familiar and available imaging modality for the evaluation of patients with acute abdominal distress. Potential causes for acute abdominal distress include the hepatobiliary system, spleen, urogenital tract, and gastrointestinal tract. Radiographic signs associated with specific conditions are described, including gastric-dilation volvulus, urinary bladder rupture, ureteral rupture, urethral rupture, pancreatitis, and small intestinal obstruction. Additionally, contrast procedures that can be beneficial in evaluating the patient with acute abdomen, including positive contrast cystography, urethrography, excretory urography, and peritoneography, are described. PMID- 12772866 TI - Ultrasonography of the acute abdomen. AB - For many years, ultrasonography has been considered to be one of the most valuable imaging techniques for evaluation of the abdominal cavity and its organs. In most cases, abdominal ultrasonography helps obtain information that leads to a definitive diagnosis. Ultrasonography is also valuable to narrow the list of differential diagnoses obtained with other diagnostic techniques. This article discusses the role of ultrasound for the diagnosis of the most common diseases that can produce clinical signs of acute abdominal pain. Abdominal organs that can be evaluated using ultrasound include the liver, biliary system, pancreas, stomach, small intestine, kidney, urinary bladder, uterus, and prostate. Pathologies of the abdominal cavity and retroperitoneal space can be also diagnosed with ultrasonography. Interventional ultrasound techniques are useful to either assist in getting the definitive diagnosis or to treat certain pathologic conditions. PMID- 12772867 TI - Abdominal paracentesis and diagnostic peritoneal lavage. AB - Abdominal paracentesis, the percutaneous removal of abdominal fluid for diagnostic and therapeutic purposes, provides a rapid, easy, and safe method of diagnosing diseases with abdominal effusion. Commonly diagnosed diseases include peritonitis, hemoperitoneum, uroabdomen, and neoplasia. Other indications for abdominal paracentesis include shock without a known apparent cause, undiagnosed disease within the abdominal cavity, suspicion of postoperative gastrointestinal wound dehiscence, blunt or penetrating abdominal injury, and refractory abdominal pain for which a cause cannot be determined. In such cases, simple abdominal paracentesis or four-quadrant paracentesis can be performed and requires minimal equipment. Diagnostic peritoneal lavage is indicated when peritonitis or other effusive disease is suspected, but other techniques have failed to provide a diagnostic sample. Both abdominal paracentesis and diagnostic peritoneal lavage are effective methods for the early detection of disease before overt clinical signs are present. PMID- 12772868 TI - Cytology and fluid analysis of the acute abdomen. AB - In patients with acute abdominal pain, abdominal paracentesis and diagnostic peritoneal lavage often yield fluid samples for cytologic and biochemical evaluation. Cytology of the effusion from a patient with acute abdominal disease can be a crucial tool for the rapid diagnosis necessary for initiation of timely and appropriate therapy. Appropriate sample collection, handling, and preparation are essential to obtain an accurate diagnosis. Analysis of the fluid sample should include gross examination of the effusion, measurement of total nucleated cell count, packed red blood cell volume, and protein concentration, as well as examination for the presence of other cells, bacteria, food particles, or plant material. Biochemical evaluation should proceed based on the clinician's index of suspicion for a particular disease process. Abdominal effusions are generally classified as transudate, modified transudate, or exudate, depending on the total nucleated cell count and protein concentration. Cytology of all fluids collected should be performed systematically, utilizing progressively higher magnifications with a microscope. Specific diseases with associated abdominal effusions include septic peritonitis, nonseptic peritonitis, hemoabdomen, uroabdomen, pancreatitis, bile peritonitis, chylous effusion, and neoplasia. A complete description of sample preparation and evaluation is reviewed. PMID- 12772870 TI - The acute abdomen: a surgeon's approach to diagnosis and treatment. AB - Clinical approach to a patient with acute abdominal distress involves decision making: whether to take a patient to surgery, or manage the patient medically. A thorough and systematic approach requires the use of diagnostic imaging modalities, including radiology and ultrasonography, performing diagnostic peritoneal lavage or abdominal paracentesis techniques, and bloodwork evaluation, including complete blood count and serum biochemistry profiles. In some cases, the results of diagnostic tests may lead to surgical versus medical management, particularly when a patient fails to respond to medical management alone. In other cases, such as GDV syndrome, penetrating abdominal wounds, pyometra, or uncontrolled abdominal hemorrhage, rapid surgical management is necessary for patient survival. This article describes a systematic approach to a small animal patient with acute abdomen. PMID- 12772869 TI - Anesthesia for the acute abdomen patient. AB - Patients with acute abdomen often have marked physiologic and pathologic changes, making anesthesia both challenging and potentially hazardous for the patient. A thorough understanding of the pathophysiologic mechanisms of cardiovascular function under anesthesia and selection of appropriate anesthetic protocols are critical to a successful anesthetic outcome. The goal is to produce anesthesia while minimizing depression of the cardiovascular system. Monitoring and management of acid-base and cardiovascular function serve to ensure appropriate oxygen delivery to the tissues during anesthesia. Postoperative management can significantly influence patient outcome following anesthesic recovery, and must therefore be considered in the anesthetic plan. Finally, pain management in all patients is an important aspect of case management, and should not be overlooked. This article serves to educate the clinician in the above-described areas in regard to the acute abdomen patient. PMID- 12772871 TI - Men and women at war. PMID- 12772873 TI - Cement pressurization after provisional repair of femoral cortical defects. AB - Cortical defects are common and problematic in cemented revision hip arthroplasty. Extruded cement can cause thermal injury, pain, and impingement. Decreased cement pressure limits bony interdigitation and leads to loosening. Historically, surgeons have used a finger to contain cement and improve pressure, and decrease porosity, but, with large or multiple defects, fingers are ineffective. Novel solutions--such as wrapping foil suture packaging or half a syringe barrel around the defects--have been previously published. In the study reported here, we used modern cementing techniques, continuous pressure monitoring, and porosity calculations to analyze the utility of the 3 provisional defect-fixation techniques. The foil and the hemisyringe worked as well as a finger (P > .05). All 3 techniques enhanced pressurization and maintained the porosity reduction. Although manually pressurizing cement and feeling resistance provide the surgeon with more tactile feedback, using the gun and a proximal adapter was more effective in improving pressure. Using these provisional defect fixation techniques as well as a cement gun and proximal adapter can improve cement pressure and decrease porosity. These techniques are particularly useful with large or multiple cortical defects encountered in revision arthroplasty or total hip arthroplasty after open reduction. PMID- 12772872 TI - Osteopetrosis. AB - Osteopetrosis is a rare skeletal condition characterized by skeletal sclerosis caused by aberrant osteoclast-mediated bone resorption. Three clinically distinct forms of osteopetrosis are recognized--the infantile malignant autosomal recessive form, the intermediate autosomal recessive form, and the adult benign autosomal dominant form. The disease represents a spectrum of clinical variants because of the heterogeneity of genetic defects resulting in osteoclast dysfunction. The pathogenic defects may be intrinsic to either the osteoclast monocyte lineage or the mesenchymal cells that constitute the microenvironment that supports osteoclast ontogeny and activation. Implicated factors include specific proto-oncogenes, growth factors, and immune regulators. A subset of patients with the intermediate autosomal recessive form has been characterized with carbonic anhydrase II isoenzyme deficiency. Management of patients with osteopetrosis requires a comprehensive approach to characteristic clinical problems including hematologic and metabolic abnormalities, fractures, deformity, back pain, bone pain, osteomyelitis, and neurologic sequelae. Medical treatment of osteopetrosis is based on efforts to stimulate host osteoclasts or provide an alternative source of osteoclasts. Stimulation of host osteoclasts has been attempted with calcium restriction, calcitrol, steroids, parathyroid hormone, and interferon. Bone marrow transplant has been used with cure for infantile malignant osteopetrosis. As osteopetrosis likely represents a spectrum of underlying etiologies resulting in osteoclast dysfunction, effective therapies most likely need to be individualized. PMID- 12772874 TI - Bilateral knee and bilateral elbow osteochondritis dissecans. AB - A 12 1/2-year-old white girl presented with a 3-year history of bilateral knee and bilateral elbow pain. Radiographs showed osteochondritis dissecans (OCD) lesions of both patellas and bilateral intra-articular bodies within the elbows- consistent with previous OCD lesions. After unsuccessful conservative management, an arthroscopy was performed on the right knee; the lesion was removed, and a chondroplasty was performed on the patella. There were no operative complications, and the patient did well postoperatively. At 12-week follow-up, a 2+ effusion was found in the right knee. Locking and pain in the left elbow were treated with arthroscopy/arthrotomy, and the 3 loose osteochondral bodies were removed. The patient regained full, painless range of motion of the left elbow. This is the first report of OCD lesions located on both patellas and elbows in an adolescent. PMID- 12772875 TI - Migration of a Steinmann pin from the proximal humerus to juxtapose the axillary artery. AB - Cases of hardware migration may be rare, but consequences of migration can be disastrous. We present a case of pin migration from the proximal humerus as a reminder of the potential complications involving use of hardware around the shoulder. PMID- 12772876 TI - Postfracture extensor pollicis longus tenosynovitis and tendon rupture: a scientific study and personal series. AB - Rupture of the extensor pollicis longus (EPL) tendon after a distal radius fracture is an uncommon event; the incidence is 3%,according to a review of treatment of 200 consecutive patients with Colles fractures. Diagnosis is based on persistent dorsal wrist pain and a positive retroflexion sign. Recommended treatments in the prerupture setting include a third dorsal compartment release with or without an extensor retinacular patch graft. Also recommended are a palmaris longus graft in the acute rupture setting and a transfer from the extensor indicis proprius to the EPL tendon in the subacute or chronic setting. Results of all treatments seem to be clinically satisfactory. PMID- 12772877 TI - Shoulder pain and the overhand athlete. AB - There has been much controversy surrounding the issue of shoulder pain and the overhand athlete. Several hypotheses have been suggested as to the cause of the shoulder pain--subacromial impingement, anterior capsular laxity, internal or posterosuperior impingement, traction injury, and scapular imbalance. Treatment for the patient depends on the specific cause of the pain. As always, a full routine of nonoperative therapeutic measures should be administered before resorting to surgical intervention. PMID- 12772878 TI - Lateral elbow ecchymosis as a clinical sign of lateral humeral condylar fractures. AB - Given the appropriate clinical history and mechanism of injury, the presence of localized lateral elbow ecchymosis in a young child is usually the sign of lateral condyle fracture of the humerus. Recognition of this clinical sign helps in directing the diagnostic studies and empiric treatment toward a lateral condyle fracture of the humerus not apparent on initial radiographs. PMID- 12772879 TI - Challenge and opportunities in the treatment of gastric cancer. PMID- 12772880 TI - The epidemiology of gastric cancer. AB - Gastric cancer mortality has declined markedly around the world. In South Australia, the reduction approximated 40% over the last 20 years. Possible reasons include: better refrigeration; reduced consumption of salted, smoked, and chemically preserved foods; increased intake of fruit and vegetables; and improved living standards and a greater use of antibiotics, which may have reduced Helicobacter pylori infection. Reductions generally have been greater for intestinal than diffuse histopathologies. Gastric cancer remains the second leading cause of cancer death worldwide, probably accounting for about 10% of newly diagnosed cancers. High rates apply to Japan, China. Central and South America, Eastern Europe, and parts of the Middle East, and low rates to North America, Australia and New Zealand, Northern Europe, and India. Rates usually are higher in lower socioeconomic groups. Five-year relative survivals of around 20% or less are frequently reported. A figure of 50% or more has been cited for Japan, where there has been radiological screening, although this exceptional figure could have been affected artificially by lead-time and related effects. Male-to-female incidence ratios generally are in the 1.5-2.5 range, with higher ratios for intestinal than diffuse cancers and higher-risk populations. In South Australia, the ratio has been 1.8 to one, although higher at 4.6 to one for cardia lesions. Recent increases in cardia cancers, especially in males in populations of European extraction, often are accompanied by increases for esophageal adenocarcinoma. It is estimated that the global burden of gastric cancer could be reduced by up to 50% by dietary changes that included an increased intake of fruit and vegetables. PMID- 12772881 TI - The role of surgery in the current treatment of gastric carcinoma. AB - Surgery currently is the only curative option in the treatment of gastric cancer. For early gastric cancer, an endoscopic mucosal resection (EMR) is adequate for intramucosal cancer less than 2 cm in diameter without ulcer. For early cancers ineligible for EMR, limited surgical operation (proximal gastrectomy, segmental resection, and pylorus-preserving distal gastrectomy) can be recommended to reduce surgical risk and achieve improvements in quality of life without decreasing survival. Subtotal/total gastrectomy plus D2 lymph node dissection is the standard surgery for advanced gastric cancer in Japan. Pancreas-preserving total gastrectomy is recommended due to the reduced risk of pancreatic fistula and postoperative diabetes. Regarding extended surgery, results of a phase III study to evaluate the role of paraaortic node dissection will be analyzed in a few years' time after the accrual of more than 500 patients in a Japan Clinical Oncology Group (JCOG) study. For scirrhous gastric cancer, left upper abdominal exenteration appears to be associated with improved survival and should be tested in another controlled trial. PMID- 12772882 TI - The role of chemotherapy in the current treatment of gastric cancer. AB - Metaanalyses of adjuvant systemic chemotherapy in gastric cancer have shown at best marginal benefits, although a recent trial of postoperative chemoradiotherapy has demonstrated significantly increased overall survival. Neoadjuvant chemotherapy and chemoradiotherapy are also feasible and are the subject of several ongoing studies. This setting may provide valuable evidence of the activity of new agents and the chemosensitivity of specific tumors. In metastatic disease, chemotherapy confers benefit when compared with best supportive care alone. No particular regimen has been shown to be superior to others in this context, and there is a pressing need for the evaluation of newer agents such as the oral fluoropyrimidines, taxanes, irinotecan, and oxaliplatin. PMID- 12772884 TI - European experience of docetaxel and cisplatin in advanced gastric cancer. AB - The combination of docetaxel with cisplatin in gastric cancer is a promising development. In a phase II study, 85-100 mg/m2 docetaxel plus 75 mg/m2 cisplatin was established as an active regimen in advanced gastric cancer (with an overall response rate of 56%) with a manageable safety profile. Up to 300 mg/m2 5 fluorouracil (5-FU) given by continuous infusion for 2 of 3 weeks can be added to this regimen without an increase in appreciable toxicity. The efficacy of docetaxel-based regimens remains to be established by a randomized phase III study. However, the results of such trials are eagerly awaited, as are data from studies of docetaxel-based combinations in the adjuvant and neoadjuvant settings. PMID- 12772883 TI - Docetaxel and cisplatin in patients with advanced gastric cancer: results of Japanese phase I/II study. AB - Two phase II monotherapy studies of docetaxel in patients with advanced gastric cancer were performed in Japan. In group A, docetaxel showed an overall response rate of 23.7% (14/59) in 59 patients, and the adverse reactions were tolerable. In group B, this agent showed the same response rate, of 23.7% (14/59) and the same adverse reaction profile. We then conducted the phase II studies of docetaxel and cisplatin for advanced gastric cancer. A Japanese phase I study in patients with advanced gastric cancer established that docetaxel 60 mg/m2 could be given together with cisplatin 80 mg/m2 without any dose-limiting toxicity. In a phase II trial of this combination regimen in 25 evaluable patients with advanced or recurrent disease, the overall response rate was 28% (7/25), and the response rate for liver metastases was 40% (6/15). Hematological and nonhematological toxicities were acceptable. It is concluded that this regimen is feasible and might warrant further investigation in respect to its relatively high response rate for liver metastases. PMID- 12772885 TI - Docetaxel in combination for advanced gastric cancer. AB - Docetaxel is considered to be active in untreated and previously treated patients with gastric carcinoma. In a multinational phase II trial (TAX 325), 158 untreated patients with advanced gastric cancer (99% without prior chemotherapy) were randomized to receive, every 3 weeks, either docetaxel 85 mg/m2 plus cisplatin 75 mg/m2 (TC) or docetaxel 75 mg/m2 plus cisplatin 75 mg/m2, plus a 5 day continuous infusion of 750 mg/m2 5-fluorouracil (FU; TCF). By intent-to-treat analysis, patients receiving TCF had a significantly higher response rate and longer time to progression. Overall survival in the two arms was not significantly different. Toxicity (particularly gastrointestinal toxicity) was greater with the TCF combination than in the TC arm, and there was a greater need for dose reduction. However, adverse events in both arms were manageable and there were no deaths associated with either regimen. Following these findings, a phase III trial comparing a control arm of cisplatin plus 5-FU against an experimental arm consisting of the TAX 325 phase II docetaxel/cisplatin/5-FU regimen is now in progress. PMID- 12772887 TI - Diamond will shine brightly for chemistry. PMID- 12772888 TI - Anionic triazacyclononanes: new supporting ligands in main group and transition metal organometallic chemistry. AB - In the course of developing new ligands to support chemistry with main-group, transition and lanthanide elements, a number of research groups have focused attention on functionalized triazacyclononanes; this article provides a summary of the more recent findings with an emphasis on the organometallic chemistry of one particular class of tacn ligands, namely those involving the anionic tacn moiety. PMID- 12772886 TI - Advances in the therapy of gastric cancer. AB - Many issues remain unclear in the management of gastric cancer. Randomized trials have failed to show the superiority of D2 over D1 dissection, and comparisons between countries showing higher survival rates following more extensive surgery may be influenced at least in part by the fact that D1 dissection underestimates disease stage in many patients. No studies have yet shown a benefit from adjuvant chemotherapy. However, the Southwest Oncology Group (SWOG) 9008 trial provides convincing evidence that a regimen of postoperative 5-fluorouracil (5-FU)-based chemoradiotherapy improves disease-free and overall survival when compared with observation alone. In the chemotherapy of advanced disease, use of the epirubicin plus cisplatin plus 5-FU (ECF) regimen leads to significantly longer median survival than 5-FU plus adriamycin plus methotrexate-C (FAMTX), but the rate of complete response remains low and the effect on longterm survival minimal. There are indications that neoadjuvant chemotherapy may increase the resectability of tumors and reduce risk of postoperative recurrence. Substantial improvements in outcome are likely to depend on the integration into multimodality strategies of novel, molecularly targeted agents in all stages of gastric cancer treatment. PMID- 12772889 TI - Facile resolution of constrained geometry indenyl-phenoxide ligation. AB - The 2-(inden-3-yl)phenoxide ligand can be resolved at both tetrahedral and octahedral Group 4 metal centers using chiral binaphthoxide ligands. PMID- 12772890 TI - Coordination polymers based on square planar Co(II) node and linear spacer: solvent-dependent pseudo-polymorphism and an unprecedented interpenetrating structure containing both 2D and 3D topological isomers. AB - Reaction of Co(NCS)2 with 1-methyl-1'-(4-pyridyl)-2-(4-pyrimidyl)ethylene (mppe) in different solvents yields two kinds of novel coordination polymer structures of [Co(mppe)2(NCS)2]n: one of them shows an unprecedented interpenetrating structure with both 2D and 3D topological isomers. PMID- 12772892 TI - Dispersing palladium nanoparticles using a water-in-oil microemulsion- homogenization of heterogeneous catalysis. AB - Palladium nanoparticles dispersed by a water-in-oil microemulsion are very effective catalysts for hydrogenation of olefins in an organic solvent. PMID- 12772891 TI - Electrical property and water repellency of a networked monolayer film prepared from Au nanoparticles. AB - Gold nanoparticles, modified with alkyl thiol, formed a film on polystyrene substrate, and it was found that the deposited film drastically changes its conductivity and hydrophobicity, depending on the alkyl chain length of the thiol used. PMID- 12772893 TI - Silica coated fullerenols: seeded growth of silica spheres under acidic conditions. AB - Liquid phase deposition of silica in the presence of fullerenol C60(OH)n, results in the formation of uniform silica spheres, whereas the use of C60 gives large non-uniform agglomerates as a result of homogeneous nucleation. Raman and UV spectroscopy indicate the C60 is retained as the core of the silica spheres. PMID- 12772894 TI - Cyclodehydrogenation of di- and tetra(benzimidazol-2-yl)benzenes to give model heteroaromatic discotic systems. AB - Di-and tetra(benzimidazol-2-yl)benzenes upon oxidation undergo cyclodehydrogenation with formation of N-N bonds to form planarized polycyclic compounds which are models for the cores of heteroatom-containing discotic materials, and which can be readily reduced back to the original compounds, thus demonstrating a molecular redox switch. PMID- 12772895 TI - Palladium catalysed cyclisation-carbonylation of enynes to give cyclic gamma,delta-unsaturated acids. AB - In the presence of acetic acid, trifurylphosphine and CO (2 atm), palladium catalyses the conversion of a range of enynes to cyclic delta,gamma-unsaturated carboxylic acids in good yield. PMID- 12772896 TI - Non-aggregating octasubstituted dendritic phthalocyanines. AB - Eight poly(aryl) ether dendritic wedges attached to the periphery of a phthalocyanine significantly decreases self-association of these chromophores in solution and the condensed state. PMID- 12772897 TI - Synthesis of ITQ-21 in OH- media. AB - ITQ-21 has been synthesized from F- free gels; aluminum can be incorporated and this does not affect the rate of nucleation. The introduction of stoichiometric amounts of NH4F with respect to the number of double four rings (D4R) present in the structure leads to an increase in the nucleation and crystallization rate with a quantitative incorporation of fluoride ions into the solid. PMID- 12772898 TI - Inhibited chelation in the new gamma-phosphino-beta-diketiminate to give phosphine --> arsine coordination. AB - A gamma-diphenylphosphino-beta-diketimine has been synthesised and the 31P NMR handle allows for in situ analysis of reaction mixtures revealing unprecedented reactivity with AsCl3 to give a novel phosphinoarsine intramolecular coordination complex. PMID- 12772899 TI - The first non-acid catalytic synthesis of tert-butyl ether from tert-butyl alcohol using ionic liquid as dehydrator. AB - Methyl tert-butyl ether (MTBE), ethyl tert-butyl ether (ETBE), and isopropyl tert butyl ether (IPTBE) have been synthesized for the first time over a non-acid ionic liquid as catalyst and dehydrator with high conversion (> 90%) and selectivity (> 90%) under mild conditions. PMID- 12772900 TI - Micropatterns constructed from Au nanoparticles. AB - Covalently linked Au-NPs micropatterns have been successfully fabricated from the self-assembly film composed of 4-mercaptophenol-capped Au nanoparticles (Au-NPs) and -N2+ containing polymers of nitro-diazoresin (NDR) by selective exposure to UV light and development in sodium dodecyl sulfate (SDS) aqueous solution. The resultant well-defined micropatterns were characterized with AFM and XPS. PMID- 12772901 TI - Highly efficient epoxidation of cyclic alkenes catalyzed by ruthenium complex. AB - The epoxidation of cyclic alkenes with molecular oxygen was efficiently completed in excellent epoxide yield using a novel ruthenium complex as catalyst under mild reaction conditions. PMID- 12772902 TI - Electrocatalytic dimerisation of non-heteroatom-substituted manganese alkynylcarbene complexes. AB - Upon controlled potential electrolysis (-1.30 V vs. SCE), the alkynylcarbene complexes MeCp(CO)2Mn=C(R)C [triple bond]CR' are seen to undergo an electrocatalytic dimerisation by coupling of the remote alkynyl carbon atoms to afford the ene-diyne dimanganese complexes [MeCp(CO)2Mn]2[eta 4-(E)-RC [triple bond] C(R')C=C(R')C [triple bond] CR]. PMID- 12772903 TI - Inner C-cyanide addition and nucleophilic addition to Ni(II) N-confused porphyrins. AB - Inner C-cyanide addition and subsequent addition of a methoxy group were observed in the reactions of Ni(II) N-confused tetra(p-tolyl)porphyrin with sodium methoxide and DDQ. PMID- 12772904 TI - Stereoselective formation of dinuclear complexes with anomalous CD spectra. AB - A novel chiral tetradentate Schiff base ligand forms dinuclear lambda,lambda [M2L2Cl2]2+ complexes with high diastereoselectivity; the complexes exhibit anomalous CD spectra due to exciton coupling of chromophores located on different metal centres. PMID- 12772905 TI - Insertion of organoindium carbenoids into rhodium halide bonds: revisiting a classic type of transition metal-group 13 metal bond formation. AB - Insertion of the carbenoid group 13 metal species InCp* (Cp* = pentamethylcyclopentadienyl) and InC(SiMe3)3 into the Rh-Cl bonds of [[RhCp*Cl2]2] yields the new complexes [Cp*Rh(InCp*)3(Cl)2] 1 and [Cp*Rh(mu2 Cl)2(InC(SiMe3)3)3] 2, respectively, exhibiting novel cage-like intermetallic complexes with In-Cl-In bridges. PMID- 12772906 TI - Photoluminescent supramolecular networks from metal-mediated assembly of polythia conjugated dieneyne. AB - Polythia conjugated dieneyne 1,1,2,5,6,6-hexakis(phenylthio)-1,5-diene-3-yne and its one- and two-dimensional silver(I) coordination polymers, which exhibited fluorescence and redox properties, were prepared and investigated. PMID- 12772907 TI - Remarkable effects of counter ions on scandium ion-promoted electron transfer reactions. AB - Scandium ion-promoted electron transfer reactions of p-benzoquinone are remarkably accelerated when tetrakis(pentafluorophenyl)borate anion is used instead of trifluoromethanesulfonate anion as the counter anion of scandium ion. Only a catalytic amount of scandium borate salt (Sc[B(C6F5)4]3) accelerates significantly the Diels-Alder reaction of 9,10-dimethylanthracene with p benzoquinone, which proceeds via Sc(3+)-promoted electron transfer from the anthracene to p-benzoquinone. PMID- 12772908 TI - Photomediated asymmetric synthesis of (-)-cuparene. AB - Generation of a benzylic quaternary stereocentre via the photomediated cyclisation of a chiral alpha-(aminobutyl)styrene followed by a microwave assisted Cope elimination has led to a total synthesis of the sesquiterpene (-) cuparene. PMID- 12772909 TI - Porphyrin hetero-dimer as charge separating system for photocurrent generation. AB - Introducing a porphyrin bearing electron acceptor onto a self-assembled monolayer (SAM) using a supramolecular method to form a hetero-dimer increased the photocurrent value compared with using porphyrin without an electron acceptor. PMID- 12772910 TI - A two-step field-induced magnetic transition in a novel layered cobalt diphosphonate. AB - Compound Na6Co7(hedp)2(hedpH)4(H2O)4 x 8H2O (hedp = 1 hydroxyethylidenediphosphonate) with a novel layer structure is reported in this paper. The layer maybe viewed as made up from two types of chains each of composition Co3(hedp)2 and Co2(hedpH)2. Magnetic measurements at 1.8 K reveal a metamagnetic transition at ca. 6.5 kOe, followed by a second transition to a ferromagnetic state at ca. 30 kOe. PMID- 12772911 TI - Preparation of ZnO nanocrystals via ultrasonic irradiation. AB - A simple and rapid process has been developed for the preparation of nanometer sized ZnO crystals via ultrasonic irradiation, by which pure ZnO nanocrystals with an average size of 6 nm and narrow size distribution can be synthesized in a short time and without using any solvents for the precipitation of ZnO. PMID- 12772912 TI - Opal gel templated synthesis of oblate titania opal materials. AB - Ordered arrays of oblate titania bodies were template synthesized using opal gel templates under uniaxial compression at ambient temperature during the favorable titania sol/gel process. The aspect ratio was controllable by the compression degree. PMID- 12772913 TI - Highly regio- and stereoselective four-component iodoamination of Se-substituted allenes. An efficient synthesis of N-(3-organoseleno-2-iodo-2(Z)-propenyl) acetamides. AB - Z-selectivity was observed for iodohydroxylation of Se-substituted allenes with I2 and H2O, which is opposite to that of 1,2-allenyl sulfoxides. With n-hexane as the co-solvent Z-iodoamination leading to N-(3-organoseleno-2-iodo-2(Z) propenyl)acetamide was observed. A brief rational for the stereoselectivity of this reaction is provided. PMID- 12772914 TI - Optical and electrical properties of the wide gap, n-type semiconductors: ZnBi2O6 and MgBi2O6. AB - Characterization of polycrystalline samples of the trirutile oxides ZnBi2O6 and MgBi2O6 reveals temperature independent conductivity (0.4 and 0.01 S cm(-1)), a negative Seebeck coefficient (-0.035 and -0.025 mV K(-1)), and an optical band gap that falls at the low energy end of visible region (1.7 and 1.8 eV), this combination of attributes, indicating that these compounds are degenerate n-type semiconductors, has not previously been observed in a Bi(5+) oxide. PMID- 12772915 TI - Fraction of the CoMoS phases accessible to NO in Co-Mo hydrodesulfurization catalysts. AB - It is established by using Co-Mo model sulfide catalysts, XAFS and FTIR that Co atoms constituting CoMoS phases are not oxidized by NO adsorption and that only 55% of the CoMoS phases is susceptible to NO adsorption even at the maximum coordinative unsaturation attainable under usual HDS reaction conditions (623-673 K). PMID- 12772916 TI - Stereoselective fluorescence quenching by photoinduced electron transfer in naphthalene-amine dyads. AB - Intramolecular chiral recognition in electron-transfer-induced fluorescence quenching has been observed for diastereomeric dyads composed of a naphthalene chromophore and an amine. PMID- 12772917 TI - Zn-proline catalyzed direct aldol reaction in aqueous media. AB - Zn complexes of proline, lysine and arginine are efficient catalysts for the aldol addition of p-nitrobenzaldehyde and acetone in aqueous medium, giving quantitative yields and enantiomeric excesses up to 56% with 5 mol% of the catalysts at room temperature. PMID- 12772918 TI - Synthesis of a eta 2-2,3-diphosphabutadiene complex of zerovalent platinum from the corresponding eta 2-phosphaalkyne complex. AB - Hydrozirconation of the eta 2-phosphaalkyne complex [Pt(dppe)(eta 2-tBuCP)] with [ZrHCl(eta 5-C5H5)2], followed by treatment with the chlorophosphaalkene ClP=C(SiMe3)2 affords the eta 2-2,3-diphosphabutadiene complex [Pt(dppe)(eta 2 tBuC(H)=PP=C(SiMe3)2]. In the presence of [Pt(PPh3)2] the latter undergoes an addition reaction with water to afford the structurally characterised Pt(II) complex [Pt(dppe)(tBuCH2P(O)HPC(SiMe3)2]. PMID- 12772919 TI - A simple and efficient method for synthesis of 5-substituted 2'-deoxyuridine nucleosides using metal-halogen exchange reaction of 5-iodo-2'-deoxyuridine sodium salt. AB - Treatment of the sodium salt of 2'-deoxy-3', 5'-bis-O-(tert-butyldimethylsilyl)-5 iodouridine (3) with n-BuLi effected regioselective lithiation at the 5-position and the following reaction with various electrophiles afforded 5-substituted 2' deoxyuridines including 1b, the precursor of stable spin-labeled 1a, in good yields. PMID- 12772920 TI - Synthesis of meso-beta doubly linked porphyrin tapes. AB - The oxidation of 5,15-bis(3,5-di-tert-butylphenyl) Ni(II)-porphyrin 1b with Sc(OTf)3 and DDQ led to production of meso-beta doubly linked Ni(II)-porphyrin tapes that have large pi-electronic communications over the arrays. PMID- 12772921 TI - Ni(III) vs. Ni(II)-thiyl radical: charge-delocalisation in a binuclear Ni(III)Ni(II)-dithiolate complex. AB - Multi-frequency EPR spectroscopy on 61Ni-labelled samples of [Ni2(L)]3+ confirms extensive charge-delocalisation between the Ni(III) centre and thiolate donors in the Ni(II)Ni(III) complex. PMID- 12772922 TI - Chiral Co(III)(salen)-catalysed hydrolytic kinetic resolution of racemic epoxides in ionic liquids. AB - In the chiral Co(III)(salen)-catalysed HKR of racemic epoxides, in the presence of ionic liquids, Co(II)(salen) complex is oxidised without acetic acid to catalytically active Co(III)(salen) complex during reaction and, moreover, this oxidation state is stabilised against reduction to Co(II) complex which enables the reuse of the recovered catalyst for consecutive reactions without extra reoxidation. PMID- 12772923 TI - Direction control in DNA binding of chiral D-lysine-based peptide nucleic acid (PNA) probed by electrospray mass spectrometry. AB - The DNA binding abilities of peptide nucleic acids (PNAs), both achiral and bearing three adjacent D-lysine-based monomers in the middle of the strand ("chiral box" PNA), were studied by means of electrospray mass spectrometry (ESI MS). In contrast with achiral PNA, "Chiral box" PNA was confirmed to exert high direction control (antiparallel vs. parallel DNA target) in DNA binding. PMID- 12772924 TI - Poly(9,9'-spirobifluorene-manganese porphyrin): a new catalytic material for oxidation of alkenes by iodobenzene diacetate and iodosylbenzene. AB - Anodic oxidation of tetraspirobifluorene-manganese porphyrin lead to the coating of the working electrode by insoluble electroactive poly(9,9'-spirobifluorene manganese porphyrin) films for which electrochemical behaviour and physicochemical properties are described; these polymeric materials are able to catalyze the heterogeneous epoxidation of styrene with iodobenzene diacetate and iodosylbenzene. PMID- 12772925 TI - Potential bias in study on career choice? PMID- 12772926 TI - Appropriate use of funds. PMID- 12772927 TI - How US departments of FM conduct grand rounds. PMID- 12772928 TI - Web site disseminates information about sun protection program. PMID- 12772929 TI - The preceptor in the 21st century. PMID- 12772930 TI - The BELIEF Instrument: a preclinical teaching tool to elicit patients' health beliefs. AB - PURPOSE: The BELIEF Instrument is a cultural interviewing tool for preclinical medical students that does not require diagnostic or therapeutic skills. METHODS: An expert panel developed and taught the instrument to 200 first-year medical students in (1) a didactic session, (2) standardized patient interviews, and (3) clinical correlation sessions with community physicians and third-year medical students. Standardized patients evaluated students on the BELIEF questions in a graded interview. RESULTS: A total of 93.5% (range 86% to 97%) of 197 students elicited information on each of the BELIEF items. CONCLUSIONS: The BELIEF instrument works as a cultural interviewing tool. It is unknown if students' interviewing behavior generalizes to real patients in clinical settings. PMID- 12772931 TI - Surprises. PMID- 12772932 TI - Teaching the realities of child abuse. PMID- 12772933 TI - Family practice graduate preparedness in the six ACGME competency areas: prequel. AB - BACKGROUND AND OBJECTIVES: Since July 2002, family practice residency program accreditation requires evidence of teaching and assessing residents in six competency areas. This study was conducted to obtain baseline information about family practice graduates' perceptions of the importance of specific competencies and the extent to which residency training prepared them to perform skills representative of the six competency areas. METHODS: A national, cross-sectional survey was conducted of family physicians who had graduated from residency programs from 1998 to 2000. RESULTS: The response rate was 54% (n=1,228). Graduates reported the most preparation in patient care skills, followed by interpersonal and communication skills and then professionalism. The least preparation was reported for skills pertinent to practice-based learning and improvement, systems-based practice, and some areas of professionalism. CONCLUSIONS: Areas of residency education that appear to warrant improvement include education about system aspects of care, practice-based learning and improvement, and selected professionalism issues. PMID- 12772934 TI - Benchmarking the costs of residency training in family practice. AB - Financial and operational benchmarking data for family practice residency programs within the University of Washington Network were established for the year 2000. Data were systematically collected by standardized questionnaire, evaluated for quality and verified, and then analyzed. Revenues, expenses, faculty structures, productivity, and family practice center staffing models are reported, using program averages and ranges or standard deviations for individual data elements. Variations and data problems included data line definitions, difficulties obtaining data from sponsoring institutions, indirect program costs, and widely differing program structures. Limited conclusions can be made regarding "best practices," but the results contribute to the establishment of normative data for budgeting and operational evaluation of family practice programs. PMID- 12772935 TI - Task-oriented processes in care (TOPIC): a proven model for teaching ambulatory care. AB - BACKGROUND AND OBJECTIVES: The TOPIC model conceptualizes the work of family physicians as five prototypical visits (new problem, checkup, chronic illness, psychosocial problem, and behavioral change) and four major processes (physician information processing, patient-physician relationship development, integration of information and relationship, and lifelong learning). This paper describes methods for teaching this model to clerkship students and reports 4 years of outcome data. METHODS: The instruction included seminars at the beginning of the rotation, a laminated pocket card as a reminder in clinical sites, a clerkship "passport" with examples of TOPIC verbal presentations, and student self-study for the end-of-rotation Clinical Performance Examination (CPX). Evaluation outcomes include student perceptions of the seminars and student performance on the CPX. RESULTS: Students rated the usefulness of the seminars favorably. The CPX scores showed that students performed most of the tasks expected for each prototypical visit. CONCLUSIONS: Students learned the tasks presented in the seminars and demonstrated them during the CPX. The model is robust since students learned the material despite some changes in teaching methods. PMID- 12772936 TI - Methods for teaching physical examination skills to medical students. AB - BACKGROUND AND OBJECTIVES: Teaching physical examination (PE) skills is a fundamental component of medical education. However, there is little information available about how medical school faculty teach PE skills. This study surveyed PE course directors to determine how they presently teach PE skills, the methods that are perceived to be the best, and how standardized patients (SPs) are recruited and paid. METHODS: A written survey was mailed to PE course directors at US allopathic and osteopathic medical schools. RESULTS: PE course directors at 83 (58%) schools responded. Results indicate that the top three methods presently used for teaching (demonstration on an SP, practice on an SP, and practice on patients) and those perceived to be the best methods are the same but in different rank order. A significant difference was observed in the overall mean scores of the present methods versus the best methods in 8 of the 10 teaching methods; thus, instructors are not always using the best methods to teach PE. There were also differences in methods used to teach "sensitive areas" (ie, genital or breast exam). CONCLUSIONS: The highest-ranked methods for teaching PE are demonstrations and practice with SPs and practice on real patients. However, PE instructors are not always using the teaching methods they deem best. PMID- 12772937 TI - Predictors of screening for breast and colorectal cancer among middle-aged women. AB - BACKGROUND AND OBJECTIVES: Rates of breast and colorectal cancer screenings among women in the United States fall below current guidelines set forth by the American Cancer Society and Healthy People 2010. This study examined predictors of breast and colorectal cancer screening behaviors among women patients (ages 50 75) at a university-based family practice clinic. METHODS: A random sample of 600 women patients were sent a comprehensive written questionnaire assessing demographic characteristics and breast and colorectal cancer screening behaviors and beliefs via the US Postal Service. RESULTS: Rates of clinical breast examinations, mammography, fecal occult blood testing, and sigmoidoscopy/colonoscopy exceeded national and state trends. A greater proportion of women reported receiving breast cancer information as compared to colorectal cancer information. Receiving information from one's physician was significantly related to having had a clinical breast examination, a fecal occult blood test, and sigmoidoscopy/colonoscopy. Significant predictors of fecal occult blood testing included lower perceived barriers, married, receiving information from physician, and greater perceived benefits. Significant predictors of sigmoidoscopy/colonoscopy included receiving information from physician, lower perceived barriers, and advanced age. CONCLUSIONS: The family physician can play an instrumental role in encouraging patients to adhere to current breast and colorectal cancer screening guidelines. PMID- 12772938 TI - Sources of error in delayed payment of physician claims. AB - BACKGROUND AND OBJECTIVES: Our objectives were to determine the distribution of errors and estimate the magnitude of the burden of delayed payments in a large physician group practice. METHODS: A 25% random sample (n=775) was taken from all billed records of a physician group practice in the Pacific Northwest that were delayed 6 months or more as of June 30, 2001. The source and specific reasons for payment delays, as well as the amount of each unpaid invoice, were determined by electronic documentation or telephone calls to the payor. Analysis of variance was used to determine whether the amount of the invoice was associated with the source and reason of error. RESULTS: The source of delayed payments due to provider, payor, patient, and technical error were 36.1%, 28.1%, 14.5%, and 21.3%, respectively. The most-frequent reasons for delayed payment were that the provider incorrectly set up the account (15.2%), the provider did not follow up on denial (12.9%), and the payor incorrectly processed the invoice (11.6%). Analysis of variance suggested that the invoice amount was not significantly associated with the source but was significantly different across reasons for delayed payment. The potential financial impact of earlier recovery of payment was $262,270. CONCLUSIONS: In these data, provider and payor errors accounted for almost two thirds of delayed payments. The most promising avenue for providers to reduce delayed payments is by reducing their own errors. Eliminating the two most common errors would result in a more timely recovery of nearly $70,000 in revenues. PMID- 12772939 TI - Evidence-based medicine meets goal-directed health care. AB - Evidence-based medicine and goal-directed, patient-centered health care seem, at times, like parallel universes, though, at a conceptual level, they are perfectly compatible. Part of the problem is that many of the kinds of information required for decision making in primary care are often unavailable or difficult to find. Several case examples are used to illustrate this problem, and reasons and solutions are suggested. The goal-directed health care model could be helpful for directing the search for evidence that is relevant to the decisions that patients and their primary care physicians must make on a regular basis. PMID- 12772940 TI - Family medicine clinicians, teachers, and researchers touching patients' lives. PMID- 12772941 TI - Residency as a hero's quest. PMID- 12772942 TI - The legacy of a family practice. PMID- 12772943 TI - Occupational rhinitis. AB - OBJECTIVE: To define occupational rhinitis, classify its various causes, review the steps in diagnosis, and describe the nonpharmacologic and pharmacologic management principles. DATA SOURCES: A review of MEDLINE articles in English on occupation rhinitis for January 1, 1970, through December 31, 2001, was performed. In addition, references were identified from bibliographies of relevant articles and books. STUDY SELECTION: The expert opinion of the author was used to select the relevant articles for the review. RESULTS: Occupational rhinitis is the episodic, work-related occurrence of sneezing, nasal discharge, and nasal obstruction. It frequently coexists with asthma but may present alone. Occupational rhinitis can be caused by heightened olfactory awareness, nonspecific inflammation of the nose, exposure to a high concentration of irritating and soluble chemical gases, or IgE mechanisms. The history and physical examination are the most important components to the workup of the patient. A site visit to the specific work area may give helpful insights to the patients' exposure. In the case of IgE-mediated allergic occupational rhinitis, skin testing or serologic testing may be useful. Greater objectivity to the diagnosis can be obtained through nasal challenge and the rapidly developing technique of rhinomanometry. Nonpharmacologic management (environmental control) and pharmacotherapy, such as that used in allergic rhinitis, should both be instituted. CONCLUSIONS: Although it does not have the same impact as occupational asthma, occupational rhinitis causes distress, discomfort, and work inefficiency. Attention to principles of management involving nonpharmacologic and pharmacologic measures will spare the patient the symptoms of occupational rhinitis. PMID- 12772944 TI - What can we learn about asthma from studying occupational asthma? AB - OBJECTIVE: To review the clinical features and underlying mechanisms of occupational asthma in an attempt to glean insights into various other forms of asthma. DATA SOURCES: Published literature, including consensus guidelines on diagnosis and management of occupational asthma. STUDY SELECTION: This article represents a synthesis of these data sources and the opinion of the author. RESULTS: Occupational asthma may be caused by a variety of mechanisms, including both IgE-dependent and non-IgE-dependent immunological processes. IgE-dependent mechanisms are responsible for reactions to all high-molecular-weight occupational antigens and to some but not all low-molecular-weight antigens. Factors in sensitization and onset include the general genetic predisposition to make IgE and the specific responsiveness of the individual to particular allergens. Once sensitized, the main factor that influences the onset of symptoms is the degree of exposure. In general, the higher the level of exposure, the more likely the sensitized person is to develop asthma. CONCLUSIONS: Occupational asthma can be induced by a variety of agents that appear to use different mechanisms to affect the airway. Studies of the remission of occupational asthma indicate that resolution is a slow process. However, the study of occupational asthma may eventually allow us to identify treatments that will accelerate remission or induce remission in other forms of asthma. PMID- 12772945 TI - I are we closer to developing threshold limit values for allergens in the workplace? AB - OBJECTIVE: To define threshold limit values and legally binding occupational exposure limits. DATA SOURCES: Review of suitable literature. STUDY SELECTION: Studies based on detailed descriptions and/or measurements of airborne allergenic dust, total allergens, or even key allergens were selected. RESULTS: Prevalences of IgE-mediated sensitization and occupational asthma are related to the aeroallergen load in workplaces. Data to set threshold limit values for flour, latex, a-amylase, and isocyanates are already sufficiently available. CONCLUSIONS: To optimize primary prevention in workplaces,health-based occupational exposure limits should be set for major occupational allergens. PMID- 12772946 TI - Workplace irritant exposures: do they produce true occupational asthma? AB - OBJECTIVE: To describe the features of irritant-induced asthma and discuss the diagnosis in relation to differing workplace irritant exposures and symptomatic responses. DATA SOURCES: A review of MEDLINE articles on this topic from January 1, 1985, through December 31, 2001 was performed. STUDY SELECTION: The author selected relevant articles for inclusion in the review. RESULTS: Many reports indicate that unintentional high-level respiratory irritant exposures can induce the new onset of asthma. Cases that meet strict criteria for a syndrome of irritant-induced asthma, termed reactive airways dysfunction syndrome, can be diagnosed with relative certainty. Several reports of irritant-induced asthma, especially prevalence studies, have relied on historical data or have otherwise modified the reactive airways dysfunction syndrome criteria for diagnosis (eg, expanding the definition to include the symptom onset several days after exposure). Such modifications, or inclusion of cases with incomplete documentation, likely increase diagnostic sensitivity but may reduce the certainty of diagnosis for individual cases. Expanding exposure criteria to moderate or long-term low-level irritant exposures causes difficulty in excluding transient irritant exacerbation of underlying asthma or coincidental onset of asthma during working life. Although recent population studies suggest a greater relative risk of asthma in occupations with expected low-to-moderate respiratory irritant exposures, currently no objective laboratory tests exist to exclude coincidental asthma in such patients. CONCLUSIONS: Irritant-induced asthma can be produced by high-level unintentional respiratory irritant exposures at work or outside the workplace. Lower levels of exposure to respiratory irritants at work are more common, and additional studies are needed to determine the airway effects of such exposures. PMID- 12772947 TI - HLA phenotype and exposure in development of occupational asthma. AB - OBJECTIVE: To review relevant scientific literature to understand the contribution of exposure to the risk of developing occupational asthma and whether HLA class 2 molecules contribute to individual susceptibility to sensitization and asthma caused by low-molecular-weight chemicals. STUDY SELECTION: The author's expert opinion was used to select relevant articles based on systematic reviews of relevant literature. RESULTS: Studies during the past decade have shown that intensity of exposure is an important determinant of asthma induced by inhaled respiratory sensitizers, both proteins and low molecular-weight chemicals. There is evidence that HLA class 2 alleles contribute to the risk of sensitization and asthma caused by low-molecular-weight chemical sensitizers. HLA-DR3 is associated with an increased risk of developing specific IgE to trimellic anhydride (TMA) and complex platinum salts such as ammonium hexachloroplate (ACP). In those exposed to ACP and possibly also to TMA, risk is greater in those who have experienced lower intensity of exposure. CONCLUSIONS: Exposure intensity is an important determinant of sensitization and asthma caused by respiratory sensitizers. HLA class 2 alleles contribute to individual susceptibility to low-molecular-weight chemicals. For some chemicals, the contribution of HLA class 2 alleles is greater in those less exposed at work to the relevant chemical. PMID- 12772948 TI - What makes a chemical an allergen? AB - OBJECTIVE: To consider the factors that confer on chemicals the ability to cause allergic sensitization, with particular emphasis on the induction of skin sensitization. DATA SOURCES: Original and review articles available in the scientific literature. STUDY SELECTION: The expert opinion of the authors was used to select studies for inclusion in this review. RESULTS: A number of requirements must be met if a chemical is to induce skin sensitization. The most important requirements are access to the viable epidermis, the formation of stable conjugates with proteins, elicitation of cytokine production by skin cells, and the initiation of T-lymphocyte responses. In addition, qualitative aspects of induced immune responses influence the form of allergic sensitization. CONCLUSIONS: An increasingly sophisticated understanding of the factors required for the development of skin sensitization and other forms of chemical-induced allergy provides new opportunities for toxicological investigation and clinical management. PMID- 12772949 TI - Control of occupational asthma and allergy in the detergent industry. AB - OBJECTIVE: To provide an overview of how a comprehensive preclinical, clinical, and industrial hygiene program has been successfully used to control allergy and asthma to enzymes used in the detergent industry. DATA SOURCES: The author performed a PubMed and ToxLine search of English-language articles with the keywords enzymes, occupational allergy, occupational asthma, detergent, and detergent industry from January 1, 1995, to January 1, 2002. Scientific meeting abstracts, books, and industry association papers on allergy and asthma in the detergent industry were also reviewed. In addition, the practical experience of one major detergent company was included in the review. STUDY SELECTION: All published work on this topic was reviewed, and the work that discussed the key highlights of control of occupational allergy and asthma to enzymes used in the detergent industry was selected for this review. RESULTS: The detergent industry has developed guidelines for the safety assessment of enzymes, control of exposure to enzymes, and medical surveillance of enzyme-exposed workers. Because of these guidelines, occupational allergy and asthma to enzymes used in the detergent industry have become uncommon events. Cases of disease have been documented in some manufacturing sites that have had poor adherence to the guidelines. Those manufacturing sites that have adhered to the guidelines have had few cases of allergy and asthma to enzymes among exposed workers. A review of medical data from these sites has shown that workers who have developed IgE antibody to enzymes can continue to work with enzymes and remain symptom free. CONCLUSIONS: Occupational allergy and asthma to enzymes used in the detergent industry have been successfully controlled via the use of preclinical, clinical, and industrial hygiene safety programs designed to minimize sensitization to enzymes and development of disease. The basic principles of these programs can be applied to other industries where occupational allergy and asthma to proteins are common. PMID- 12772950 TI - Recent developments in diisocyanate asthma. AB - OBJECTIVE: To summarize the latest experimental findings on diisocyanate asthma and discuss the impact of these data on our understanding of disease pathogenesis and diagnosis. DATA SOURCES: The literature reviewed includes articles from PubMed (National Library of Medicine) published within the last 3 years (1999 2001). In addition, pertinent older references are discussed to provide a historical perspective and background. STUDY SELECTION: The data discussed were chosen to highlight key concepts relevant to diisocyanate asthma pathogenesis and are grouped accordingly. RESULTS: In many ways, diisocyanate-induced asthma mirrors allergic asthma caused by other stimuli; however, the immune-mediated pathways believed to be central to the disease have been difficult to define. Recent studies on the human immune response to diisocyanates provide additional evidence supportive of an immune basis for pathogenesis but also highlight well recognized differences between diisocyanate asthma and common atopic asthma. Studies on the antigenic form of diisocyanates and their interaction with epithelial tissues provide new insights that may help explain these apparent immunologic differences. Genetic factors that influence disease have begun to be identified but remain poorly characterized. Associations of particular major histocompatibility complex class II alleles with diisocyanate asthma further fuel the hypothesis that immune-dependent mechanisms underlie pathogenesis, whereas associations of glutathione S-transferase polymorphisms (in conjunction with recent studies defining the effects of diisocyanates on thiol-redox homeostasis) may implicate additional antigen-independent mechanisms. Long-term follow-up studies of diisocyanate asthma patients have confirmed the prognostic value of early removal of symptomatic patients from exposure and highlight the need for effective diagnostic tests of sensitivity and susceptibility. CONCLUSIONS: Diisocyanate-induced asthma appears to be a multifactorial disease involving the immune system, airway epithelium, and genetic factors. The potential long-term adverse effects of diisocyanate exposure in sensitized patients underscore the need for further studies to elucidate the pathogenesis of this disease and identify biomarkers for sensitization and susceptibility. PMID- 12772951 TI - Strategies for distinguishing asymptomatic latex sensitization from true occupational allergy or asthma. AB - OBJECTIVE: To evaluate strategies for distinguishing those individuals with true occupational asthma caused by exposure to natural rubber latex from those individuals with allergic disease related to other allergens. DATA SOURCES: Article published between January 1, 1981, and December 31, 2001, were identified via a MEDLINE search with the following keywords: latex, allergy, asthma, occupation asthma, and adverse reactions. STUDY SELECTION: English-language reports concerning diagnostic methods in latex and other forms of occupational asthma. RESULTS: Many methods have been evaluated for the diagnosis of latex allergy, including medical history, skin prick tests, in vitro tests, and various challenge tests. Skin prick tests with well characterized latex extracts are highly sensitive and specific predictors of latex-specific IgE antibodies; however, direct pulmonary challenge with latex allergen appears to be the only highly reliable method for diagnosis of latex-related occupational asthma. CONCLUSIONS: It is difficult to optimally distinguish between occupational latex asthma and asymptomatic latex sensitization in a person with preexisting asthma using currently available techniques. PMID- 12772952 TI - Use of pulmonary function tests in the diagnosis of occupational asthma. AB - OBJECTIVE: To discuss the different methods of assessing lung function measurements for the diagnosis of occupational asthma, focusing in particular on serial peak expiratory flow rate (PEFR) monitoring, including details on how PEFR records should be kept, plotted, and analyzed and limitations of the method. DATA SOURCES: Published studies on the use of diagnostic methods in occupational asthma, expert opinion, and recently obtained data from studies performed at a large occupational lung disease clinic. STUDY SOURCES: The expert opinion of the author was used to select the relevant data for review. RESULTS: Objective methods are necessary for the diagnosis of occupational asthma, since clinical history alone is not a satisfactory means of diagnosis. Serial PEFR monitoring has a high diagnostic sensitivity and specificity for occupational asthma and is more useful than evaluation of cross-shift change in forced expiratory volume in 1 second or change in nonspecific bronchial hyperresponsiveness. Interpretation is best performed by expert visual evaluation of plots of maximum, mean, and minimum daily PEFR readings. CONCLUSIONS: Despite some limitations of the method, serial PEFR monitoring is usually the most appropriate first-line investigation in workers suspected of having occupational asthma. PMID- 12772954 TI - Emerging research needs for membrane processes. PMID- 12772953 TI - Persistence of bronchial reactivity to occupational agents after removal from exposure and identification of associated factors. AB - OBJECTIVES: To examine the outcome of specific bronchial reactivity to occupational agents in patients with occupational asthma who have been removed from exposure to the offending agents and to identify risk factors for the persistence of specific bronchial reactivity. DATA SOURCES: A PubMed (National Library of Medicine) search of articles published from January 1, 1985, through December 31, 2001, with the keywords occupational asthma and bronchial reactivity was performed. STUDY SELECTION: This article represents a synthesis of these PubMed sources and the opinion of the author. RESULTS: Specific bronchial reactivity persists in most cases even a long time after removal from exposure to the offending agents. This condition does not seem to be correlated with the persistence of bronchial reactivity to methacholine but seems to be related to the persisting level of specific IgE to the occupational agents in the case of high-molecular-weight agents and platinum salts. CONCLUSIONS: Patients who have been diagnosed as having occupational asthma are at risk of developing an asthmatic reaction in the minutes following re-exposure to the offending agent, even if they no longer experience asthma symptoms, have normalized their bronchial reactivity, and have not been exposed to the offending agent for years. It is, therefore, crucial that these patients remain unexposed to the agent that caused their occupational asthma. PMID- 12772955 TI - Simazine and metolachlor removal by subsurface flow constructed wetlands. AB - A subsurface flow constructed wetlands system was used in this pilot-scale field study to treat runoff from a container nursery. The goal of the study was to determine the statistical significance of certain factors on the removal of simazine (2-chloro-4,6-bis[ethylamino]-s-triazine) and metolachlor (2-chloro-N-[2 ethyl-6-methylphenyl]-N-[2-methoxy-1-methylethyl] acetamide). The factors analyzed included the hydraulic flowrate, media depth, cell aspect ratio, and the presence or absence of Scirpus validus. Analysis of variance using the general linear model procedure determined that vegetation and flowrate significantly (alpha = 0.05) affected herbicide mass removal. The results showed that vegetated subsurface flow cells generally remove more simazine and metolachlor than nonvegetated cells. Decreasing the hydraulic loading rate and, therefore, the areal simazine mass loading rate to vegetated cells from 1659 to 415 mg/m2 caused an increase in the percentage of total mass removed from approximately 60 to 96%. Similarly, metolachlor mass removal percentage increased from approximately 62 to 96% when the areal mass loading to vegetated cells decreased from 1037 to 260 mg/m2. Media depth and aspect ratio did not significantly (alpha = 0.05) affect effluent simazine and metolachlor concentrations. Peak effluent metolachlor concentrations were less than 300 microg/L from vegetated cells receiving hydraulic loading rates of 204 and 102 m3/d x ha. Peak simazine concentrations in the effluent of cells receiving similar hydraulic loadings were approximately 1400 and 750 microg/L (204 and 102 m3/d x ha), respectively. PMID- 12772956 TI - Preferential transport of Cryptosporidium parvum oocysts in variably saturated subsurface environments. AB - When oocysts of the protozoan Cryptosporidium parvum contaminate drinking water supplies, they can cause outbreaks of Cryptosporidiosis, a common waterborne disease. Of the different pathways by which oocysts can wind up in drinking water, one has received little attention to date; that is, because soils are often considered to be perfect filters, the transport of oocysts through the subsoil to groundwater is generally ignored. To evaluate the significance of this pathway, a series of laboratory experiments investigated subsurface transport of oocysts. Experiment 1 was carried out in a vertical 18-cm-long column filled either with glass beads or silica sand, under conditions known to foster fingered flow. Experiment 2 involved undisturbed, macroporous soil columns subjected to macropore flow. Experiment 3 aimed to study the lateral flow on an undisturbed soil block. The columns and soil samples were subjected to artificial rainfall and were allowed to reach steady state. At that point, feces of contaminated calves were applied at the surface along with a known amount of potassium chloride to serve as a tracer, and rainfall was continued at the same rate. The breakthrough of oocysts and chloride, monitored in the effluent, demonstrate the importance of preferential flow on the transport of oocysts. Compared with chloride, peak oocyst concentrations were not appreciably delayed and, in some cases, occurred even before the chloride peak. Recovery rates for oocysts were low, ranging from 0.1 to 10.4% of the oocysts originally applied on the columns. However, the numbers of oocysts present in the effluents were still orders of magnitude higher than 10 oocysts, the infectious dose considered by the U.S. Food and Drug Administration, Center for Food Safety and Applied Nutrition, to be sufficient to cause Cryptosporidiosis in healthy adults. These results suggest that the transport of oocysts in the subsurface via preferential flow may create a significant risk of groundwater contamination in some situations. PMID- 12772957 TI - Examination of mechanisms for odor compound generation during lime stabilization. AB - Lime-stabilized biosolids produced from a wastewater treatment plant often emit odors, especially those described as "fishy" and "decaying". These odors can generate public opposition to biosolids land-application programs even though they represent an environmentally friendly recycling of organic material that is beneficial to the agricultural industry. Therefore, it is critical to examine the controlling factors involved in odor production during the lime stabilization process. Results from preliminary experiments examining added polymer and protein material to dewatered limed biosolids show increased trimethylamine (TMA) production with further increases in 1-hour and 4-hour storage times prior to liming. Further experiments with water-silica slurry reaction media reveal that enzymatically facilitated degradation of polymer and protein is the overriding factor in TMA and dimethyldisulfide (DMDS) production. It is hypothesized that macromolecules such as polymer and proteins in biosolids are first broken down enzymatically, upon which the addition of lime causes TMA and DMDS to be released. PMID- 12772958 TI - Relative efficacy of intrinsic and extant parameters for modeling biodegradation of synthetic organic compounds in activated sludge: steady-state systems. AB - The performance of intrinsic and extant kinetic parameters as predictors of synthetic organic compound (SOC) concentration in biotreatment systems operated at steady state was evaluated. Two laboratory-scale, completely mixed activated sludge systems were sampled on a routine basis, and SOC concentrations were quantified using gas chromatography with flame-ionization detection coupled with solid-phase microextraction for analyte concentration. At the same time, intrinsic and extant respirometric tests were performed periodically, and the kinetic parameter estimates obtained were used to predict effluent SOC concentrations for comparison with the measured values. Out of 28 comparisons that could be made between intrinsic and extant predictions, extant parameters were superior in 27 cases and intrinsic parameters were comparable, at best, to extant parameters in the remaining case. Given their superior performance and relative ease of measurement, extant parameters are preferable for use in design and operational decision-making. PMID- 12772959 TI - Simultaneous biological nutrient removal: evaluation of autotrophic denitrification, heterotrophic nitrification, and biological phosphorus removal in full-scale systems. AB - Simultaneous biological nutrient removal (SBNR) is the biological removal of nitrogen and phosphorus in excess of that required for biomass synthesis in a biological wastewater treatment system without defined anaerobic or anoxic zones. Evidence is growing that significant SBNR can occur in many systems, including the aerobic zone of systems already configured for biological nutrient removal. Although SBNR systems offer several potential advantages, they cannot be fully realized until the mechanisms responsible for SBNR are better understood. Consequently, a research program was initiated with the basic hypothesis that three mechanisms might be responsible for SBNR: the reactor macroenvironment, the floc microenvironment, and novel microorganisms. Previously, the nutrient removal capabilities of seven full-scale, staged, closed-loop bioreactors known as Orbal oxidation ditches were evaluated. Chemical analysis and microbiological observations suggested that SBNR occurred in these systems. Three of these plants were further examined in this research to evaluate the importance of novel microorganisms, especially for nitrogen removal. A screening tool was developed to determine the relative significance of the activities of microorganisms capable of autotrophic denitrification and heterotrophic nitrification-aerobic denitrification in biological nutrient removal systems. The results indicated that novel microorganisms were not substantial contributors to SBNR in the plants studied. Phosphorus metabolism (anaerobic release, aerobic uptake) was also tested in one of the plants. Activity within the mixed liquor that was consistent with current theories for phosphorus-accumulating organisms (PAOs) was observed. Along with other observations, this suggests the presence of PAOs in the facilities studied. PMID- 12772960 TI - High nitrite buildup during nitrification in a rotating disk reactor. AB - Incomplete nitrification with high nitrite accumulation has three practical advantages: lower oxygen consumption, less need for organics for denitrification, and lower sludge production during denitrification. Nitrification leading to high nitrite formation was experimentally studied in a continuous single rotating disk reactor (RDR) and compared to a modeled continuous completely stirred tank reactor (CSTR). The results of this model show that to accumulate nitrite greater than 50% at oxygen levels higher than 3.5 mg O2/L, pH levels higher than 8.5 and 9.0 are required for a CSTR with and without cell washout, respectively. For a CSTR without cell washout at pH 7 and 1 mg O2/L, it was predicted that a nitrite accumulation less than 5% could be reached. Conversely, for a partially submerged continuous RDR without any additional aeration supply (already at pH 7 and 1.3 mg O2/L), high nitrite accumulation (more than 75%) was achieved and the influence of pH from 7 to 9 was not significant. This difference is believed to be caused by mass transfer. In addition, nitrification was observed to occur under oxygen transport limitation for a totally submerged continuous RDR. PMID- 12772961 TI - Detection of infectious human adenoviruses in tertiary-treated and ultraviolet disinfected wastewater. AB - A pilot-scale study investigating the use of low-pressure, high-intensity UV radiation for disinfection of urban wastewater was conducted. The inactivation of coliform bacteria, wastewater-indigenous enteric viruses, seeded poliovirus, and seeded F-specific coliphage was studied. During the course of the pilot study, infectious human adenoviruses were isolated from 15 of 16 large-volume samples of UV-disinfected secondary- and tertiary-treated wastewater. Half of the tertiary treated, UV-disinfected effluent samples from which the adenoviruses were isolated had total coliform concentrations that complied with California's Water Recycling Criteria. To determine the relative UV resistance of the adenovirus isolates, purified viruses were seeded into tertiary-treated waste-water and exposed to low-pressure, high-intensity, collimated UV radiation. A dose of approximately 170 mW-s/cm2 was required to achieve 99.99% inactivation. These findings suggest that UV doses effective at meeting certain wastewater regulations for total coliform bacteria may not provide suitable inactivation of the UV-resistant human adenoviruses. PMID- 12772962 TI - Decolorization of the dye, Reactive Blue 19, using ozonation, ultrasound, and ultrasound-enhanced ozonation. AB - Reactive dyes constitute a significant portion of colorants used in industries ranging from the textile industry to the paper industry. In most cases, the effluent streams from textile plants are highly colored, and treatment methods for dye decolorization such as chemical oxidation need to be explored. The oxidation processes investigated in this study are those of ozonation, ultrasound, and ultrasound-enhanced ozonation. The oxidation of an anthraquinone dye was studied under conditions of varying ultrasonic power, dye concentration, ozone concentration, pH, and temperature. Laboratory experiments were performed using a semibatch reactor by ozonating dye samples with and without ultrasound. Under conditions of constant ultrasonic radiation and continuous gas application, decolorization rates have been enhanced by ultrasound. The apparent first-order rate constants increased between 35 and 204% for the ultrasonic power inputs between 40 W/L and 120 W/L compared with ozonation alone. The effects of ultrasonic power input on the gas-liquid mass-transfer coefficient were also investigated and the results indicate that an increase in ultrasonic power input increases the mass-transfer coefficient. The mass-transfer coefficient increased between 89 and 93% for ozone inlet concentrations between 5.4 and 9.4 mg/L at an ultrasonic power of 120 W/L compared with ozonation alone. The reactions of the dye with ultrasound-enhanced ozone occurred through the hydroxyl radical pathway. PMID- 12772963 TI - Catalytic wet oxidation: mathematical modeling of multicompound destruction. AB - A mathematical model of a three-phase catalytic reactor, CatReac, was developed for analysis and optimization of a catalytic oxidation reactor that is used in the International Space Station potable water processor. The packed-bed catalytic reactor, known as the volatile reactor assembly (VRA), is operated as a three phase reactor and contains a proprietary catalyst, a pure-oxygen gas phase, and the contaminated water. The contaminated water being fed to the VRA primarily consists of acetic acid, acetone, ethanol, 1-propanol, 2-propanol, and propionic acid ranging in concentration from 1 to 10 mg/L. The Langmuir-Hinshelwood Hougen Watson (L-H) (Hougen, 1943) expression was used to describe the surface reaction rate for these compounds. Single and multicompound short-column experiments were used to determine the L-H rate parameters and calibrate the model. The model was able to predict steady-state multicomponent effluent profiles for short and full scale reactor experiments. PMID- 12772964 TI - Pulp biology: progress during the past 25 years. AB - During the past 25 years there has been a rapid expansion in our knowledge of the dentine and pulp complex. This paper provides representative examples of important advances that researchers have made in this field. Topics to be considered include: differentiation of odontoblasts, dentine matrix proteins, extent of odontoblast processes, pulpal stem cells, apoptosis, interstitial fluid pressure in normal and inflamed pulps, class II antigen-presenting cells of the pulp, cytokines, antibodies, pulpal calcifications, tertiary dentine and pulpal inflammation associated with bacterial contamination of exposed dentine beneath restorations. PMID- 12772965 TI - To do a "root canal". AB - Endodontic treatment is often referred to as "a root canal". This article discusses the need to be more refined and descriptive in the selection of treatment alternatives. The pathological involvement of teeth needing endodontic treatment varies significantly from superficial pulp inflammation to pulp necrosis and infection, often complicated with a periradicular osteolytic process. This diversity should lead to a variety of treatment modalities based on the severity of the disease. Failure to do so and use a one-size-fits-all approach--"a root canal"--is an oversimplification and a disservice to the patient and the profession. PMID- 12772966 TI - Cytotoxicity of a new endodontic filling material. AB - In vitro cell cultures have been widely used as a means of evaluating cytotoxicity of root canal filling materials. Following ANSI/ADA spec. no. 41, the aim of the present study was to investigate the biological compatibility of a new sealer (FibreFill) and compare it with some commercially available endodontic sealers (Bioseal and Acroseal). Mouse 3T3 fibroblasts were seeded and cultured and subsequently extracts of the cements were added. After 24 hours incubation, the cellular vitality of fibroblasts was evaluated by the neutral red uptake test (NRU), which measures the membrane permeability. Data were collected and statistically analysed. Results showed that all tested materials exhibited mild cytotoxic effects, which are compatible with normal clinical use, and no statistically significant difference was noted between FibreFill and the other tested materials. Therefore, selection amongst these sealers should be based on other factors. PMID- 12772967 TI - Adhesion of a glass-ionomer root canal sealer to human dentine. AB - This in vitro study evaluated the effect of smear layer removal on adhesion to human dentine of Ketac-Endo, Grossman, and Sealer 26 root canal sealers. A total of 60 extracted human maxillary and mandibular molars with their crowns ground flat were used. The teeth were divided into two groups: Group 1, the dentine surface received no treatment; Group 2, EDTA-C was applied to the dentine surface for 5 min. Ten samples were tested for each sealer and each group. Adhesion was measured with a Universal Testing Machine. Sealer 26 showed greater adhesion both with and without smear layer (p < 0.01). Ketac-Endo and Grossman sealers were statistically equal, having the lowest values of adhesion. Application of EDTA-C did not alter the adhesion of Ketac-Endo and Grossman sealers. However, the use of EDTA-C increased the adhesion of Sealer 26. The epoxy resin-based root canal sealer (Sealer 26) adhered better to dentine prepared with and without EDTA-C than glass-ionomer-based sealers and the zinc oxide eugenol-based (Ketac-Endo and Grossman) sealers. PMID- 12772968 TI - Effective cleaning protocols for rotary niclkel-titanium files. AB - Cleaning of dental instruments before sterilisation is an essential part of processing for re-use. Rotary nickel-titanium (NiTi) files as obtained from the manufacturer are mostly not sterile and require cleaning and sterilisation both before first use and for re-use. We describe here two simple, practical cleaning protocols that are effective in removing organic debris from files. Rotary NiTi files can be safely re-used from an infection control perspective. PMID- 12772969 TI - The use of an ultrasonic technique to remove a fractured rotary nickel-titanium instrument from the apical third of a curved root canal. AB - The frequency of use of nickel-titanium rotary instruments in endodontic practice is increasing. One occasionally mentioned problem is fracture of an instrument within the root canal. The broken instrument may prevent access to the root apex, impeding optimal cleaning and shaping procedures. A case report is presented which uses a modern technique, utilising a staging platform and the use of newly designed ultrasonic tips with direct visualisation through a dental operating microscope, to remove a fractured rotary nickel-titanium instrument from the apical third of a curved root canal. PMID- 12772970 TI - Proliferative activity in periapical lesions. AB - The identification of proliferative activity in periapical lesions may be useful in discovering the biological behaviour of different cell types. Proliferant cell nuclear antigen (PCNA), Ki67, CD3 and p53 have been used for the evaluation of the proliferative ability of many lesions. In this study, 16 periapical granulomas and 8 radicular cysts were analysed. A semi-quantitative method was used for evaluation of each lesion and antigen. According to the intensity of staining the following scores were assigned: 0 = no reactivity; 1 = reactivity above background; 2 = moderate reactivity; 3 = strong reactivity. Ki67 positive cells were present in all the periapical lesions. The antibody Ki67 was positive in pathological keratinic cells, while it was only slightly positive in normal epithelial cells. PCNA positive cells were present in 22 of the 24 cases examined. CD3 reactivity was also highly positive in periapical lesions. The reaction of oncoprotein p53 in periapical lesions was negative. The expression of the positive reaction of Ki67 and PCNA in periapical lesions is therefore an indicator of cell proliferation as a result of a chronic irritative stimulus. PMID- 12772971 TI - The use and predictable placement of Mineral Trioxide Aggregate in one-visit apexification cases. AB - Endodontic treatment of the pulpless tooth with an immature root apex poses a special challenge for the clinician. The main difficulty encountered is the lack of an apical stop against which to compact an interim dressing of calcium hydroxide (Ca(OH)2), or the final obturation material. In these situations the unpredictability of the result, the difficulty in creating a leak-proof temporary restoration for the duration of treatment, and the difficulty in protecting the thin root from fracture may lead to complications when using traditional (Ca(OH)2 based) apexification techniques. Furthermore, given the increased mobility of today's society, lengthy treatment protocols are fraught with problems, and may not be followed through to completion. This may lead to ultimate failure of the case. Mineral Trioxide Aggregate (MTA) has recently been introduced for use in endodontics. Current literature supports its efficacy in a multitude of procedures including apexification. The focus of this paper is to propose a one visit apexification protocol with MTA as an alternative to the traditional treatment practices with Ca(OH)2. One-visit apexification may shorten the treatment time between the patient's first appointment and the final restoration. The importance of this approach lies in the expedient cleaning and shaping of the root canal system, followed by its apical seal with a material that favours regeneration. Furthermore, the potential for fractures of immature teeth with thin roots is reduced, as a bonded core can be placed immediately within the root canal. PMID- 12772972 TI - A comparative analysis of Mineral Trioxide Aggregate and Portland cement. AB - The purpose of this study was to compare the composition of Portland cement and Mineral Trioxide Aggregate (MTA). Samples of MTA and Portland cement were analysed for fifteen different elements by inductively coupled plasma emission spectrometry (ICP-ES). Comparative analysis revealed there was significant similarity except there was no detectable quantity of Bismuth in Portland cement. Quantitative results are given in both parts per million (p.p.m.) and wt%. It was concluded that there is no significant difference between the 14 different elements in both Portland cement and MTA. PMID- 12772974 TI - Specialist registration in Victoria--training of specialists in endodontics. AB - A historical review is presented of the growth and development of the MDSc programme in Endodontics at the University of Melbourne. PMID- 12772975 TI - Decrowning teeth and immediate extirpation of pulp. 1899 [classical article]. PMID- 12772973 TI - MTA apexification combined with conventional root canal retreatment. AB - Apexification aims to induce apical closure of the open root apex with a hard tissue barrier, against which a root filling can be compacted. Despite the popularity of the apexification procedure, calcium hydroxide therapy has some disadvantages that include variability of treatment time, unpredictability of apical closure, difficulties with patient follow-up and delayed treatment. Mineral trioxide aggregate (MTA) is a potential apical barrier material with good sealability and a high degree of biocompatibility. This paper demonstrates the placement of an apical barrier using MTA as an alternative to conventional long term calcium hydroxide therapy. PMID- 12772976 TI - A time for reflection. PMID- 12772977 TI - Compared with inverse-planning, forward planning is preferred for IMRT stereotactic radiosurgery. For the proposition. PMID- 12772978 TI - Compared with inverse-planning, forward planning is preferred for IMRT stereotactic radiosurgery. Against the proposition. PMID- 12772979 TI - Investigating the effect of cell repopulation on the tumor response to fractionated external radiotherapy. AB - In this work we study the descriptive power of the main tumor control probability (TCP) models based on the linear quadratic (LQ) mechanism of cell damage with cell recovery. The Poisson, binomial, and a dynamic TCP model, developed recently by Zaider and Minerbo are considered. The Zaider-Minerbo model takes cell repopulation into account. It is shown that the Poisson approximation incorporating cell repopulation is conceptually incorrect. Based on the Zaider Minerbo model, an expression for the TCP for fractionated treatments with varying intervals between two consecutive fractions and with cell survival probability that changes from fraction to fraction is derived. The models are fitted to an experimental data set consisting of dose response curves that correspond to different fractionation regimes. The binomial TCP model based on the LQ mechanism of cell damage solely was unable to fit the fractionated response data. It was found that the Zaider-Minerbo model, which takes tumor cell repopulation into account, best fits the data. PMID- 12772980 TI - MLC quality assurance techniques for IMRT applications. AB - Intensity modulated radiotherapy (IMRT) requires extensive knowledge of multileaf collimator (MLC) leaf positioning accuracy, precision, and long-term reproducibility. We have developed a technique to efficiently measure the absolute position of each MLC leaf, over the range of leaf positions utilized in IMRT, based on dosimetric information. A single radiographic film was exposed to 6 MV x-rays for twelve exposures: one open field with a radio-opaque marker tray present, and eleven fields (1 x 28 cm strips via 1 cm gaps between opposed leaf pairs) separated by 2 cm center to center. The process was repeated while varying direction of leaf travel; each film was digitized using a commercial film dosimetry system. The digital images were manipulated to remove translation and rotation of the film data with respect to the collimator coordinate system by extraction of radiation dose profiles perpendicular to the MLC leaf motion and measuring the center of the x-ray leakage between leaves. Radiation dose profiles in the direction of leaf motion were acquired through the center of each leaf pair (leaves 2-28), which provided leaf position information every 2 cm with 0.2 mm precision. Nine separate leaf reproducibility studies over a 90 day period which evaluated 600 measurement points on each film show 0.3 mm precision for 95% confidence, while hysteresis studies show 0.5 mm precision. Absolute leaf position error measurements demonstrated a radial dependence, with a maximum of 1.5 mm at 16.4 cm from central axis, due to rotational error at calibration. Recalibration of the MLC leaves based utilizing this tool yields absolute leaf position measurements where 91.5% of all leaves/positions were within 0.5 mm, with a mean error of 0.1 mm and a maximum error less than 1.0 mm. PMID- 12772981 TI - The use of linear programming in optimization of HDR implant dose distributions. AB - The introduction of high dose rate brachytherapy enabled optimization of dose distributions to be used on a routine basis. The objective of optimization is to homogenize the dose distribution within the implant while simultaneously satisfying dose constraints on certain points. This is accomplished by varying the time the source dwells at different locations. As the dose at any point is a linear function of the dwell times, a linear programming approach seems to be a natural choice. The dose constraints are inherently linear inequalities. Homogeneity requirements are linearized by minimizing the maximum deviation of the doses at points inside the implant from a prescribed dose. The revised simplex method was applied for the solution of this linear programming problem. In the homogenization process the possible source locations were chosen as optimization points. To avoid the problem of the singular value of the dose at a source location from the source itself we define the "self-contribution" as the dose at a small distance from the source. The effect of varying this distance is discussed. Test cases were optimized for planar, biplanar and cylindrical implants. A semi-irregular, fan-like implant with diverging needles was also investigated. Mean central dose calculation based on 3D Delaunay-triangulation of the source locations was used to evaluate the dose distributions. The optimization method resulted in homogeneous distributions (for brachytherapy). Additional dose constraints--when applied--were satisfied. The method is flexible enough to include other linear constraints such as the inclusion of the centroids of the Delaunay-triangulation for homogenization, or limiting the maximum allowable dwell time. PMID- 12772982 TI - Removal of arterial wall calcifications in CT angiography by local subtraction. AB - CT Angiography (CTA) is an established technique for the minimally invasive imaging of arteries. The technique of maximum intensity projection (MIP) is often used to get a comprehensive overview of the vascular anatomy. On a MIP, however, arterial wall calcifications may hinder the visualization of the arterial lumen. These calcifications are in direct contact with the contrast-enhanced blood, which makes removal difficult. We present a local subtraction method for the automatic removal of these calcifications. In our approach a second CT scan has to be made, prior to contrast injection. The calcifications in both scans are registered prior to subtraction to compensate for displacements in between the two scans. Local subtraction results are compared with results obtained by thresholding. The method was tested in a phantom and with data from four patients. The phantom represented an artery with different types of stenosis. Data were used from patients for which CTA of the renal arteries was performed. For two patients the electrocardiogram (ECG) was recorded during the CTA examination, making retrospective cardiac gated reconstructions possible. Both the phantom and the patient study showed that the local subtraction method is capable of removing calcifications and visualizing the residual lumen. In the patient study it appeared that some artifacts remained for higher pitch values. We conclude that the local subtraction method is less subjective and more accurate than thresholding. Best results are obtained by use of a small pitch, at the expense of the volume covered during a single breath hold. PMID- 12772983 TI - Thin CdTe detector in diagnostic x-ray spectroscopy. AB - A CdTe Schottky diode detector of 1 mm thickness was employed in diagnostic x-ray spectroscopy. The detector response to monoenergetic photons was investigated with gamma rays from the calibration sources (241Am and 133Ba). As spectral distortion due to carrier trapping, known as tailing, was small in gamma-ray spectra, the effects of carrier trapping were not taken into account in the calculation of response functions. The distortion due to the transmission of primary x rays and the escape of secondary x rays (K-fluorescent x rays and Compton-scattered x rays) from the crystal was included in the calculated response functions. X-ray spectra corrected using the response functions were in good agreement with the reference spectra obtained with a high-purity germanium detector. The results indicated that correction for the distortion due to carrier trapping is not necessary when using a thin CdTe detector in diagnostic x-ray spectroscopy. PMID- 12772984 TI - Evaluation of cost functions for gray value matching of two-dimensional images in radiotherapy. AB - In external beam radiotherapy, portal imaging is applied for verification of the patient setup. Current automatic methods for portal image registration, which are often based on segmentation of anatomical structures, are especially successful for images of the pelvic region. For portal images of more complicated anatomical structures, e.g., lung, these techniques are less successful. It is desirable to have a method for image registration that is applicable for a wide range of treatment sites. In this study, a registration method for two-dimensional (2D) registration of portal and reference images based on intensity values was tested on portal images of various anatomical sites. Tests were performed with and without preprocessing (unsharp mask filtering followed by histogram equalization) for 96 image pairs and six cost functions. The images were obtained from treatments of the rectum, salivary gland, brain, prostate, and lung. To get insight into the behavior of the various cost functions, cost function values were computed for each portal image for 20,000 transformations of the corresponding reference image, translating the reference image in a range of +/- 1 cm and rotating +/- 10 degrees with respect to the clinical match. The automatic match was defined as the transformation associated with the global minimum (found by an exhaustive search). Without preprocessing, the registration reliability was low (less than 27%). With preprocessing, about 90% of the matches were successful, with a difference with our gold standard (manual registration) of about 1 mm and 1 degree SD. All tested cost functions performed similarly. However, the number of local minima using mutual information was larger than for the other tested cost functions. A cost function based on the mean product of the corresponding pixel values had the least number of local minima. In conclusion, gray value based registration of portal images is applicable for a wide range of treatment sites. However, pre-processing of the images is essential. PMID- 12772985 TI - Accurate and efficient detection of pulmonary seed embolization in prostate iodine-125 permanent brachytherapy with a collimated gamma scintillation survey meter. AB - Pulmonary seed embolization is frequently observed in permanent prostate brachytherapy. Postoperative chest radiographic examination does not always detect seed embolization. To overcome this deficiency, a low energy gamma scintillation survey meter was converted to a seed-migration detector by adding a cone-shaped single-hole collimation cap to the window end of the scintillation probe. The response functions of the seed-migration detector to iodine-125 (I 125) for different source-to-detector distances in air and in water were measured. The spatial discrimination power of the survey meter, represented by the full width at half maximum measured in water, is typically improved from more than 7 cm to about 3 cm. Seventy-nine patients with I-125 implantation were scanned with the seed-migration detector at the patients' 30-day postevaluation visit. Fifteen patients showed single-seed embolization to the chest region and four patients displayed two-seed embolization. In other words, 24% of the patients present with embolized seeds. The detection accuracy of each patient was validated by a comprehensive investigation procedure. The comprehensive investigation consists of reviewing the patient's treatment history, orally questioning the patient for possible seed loss via the urethra route outside the hospital, examining all available chest radiographs before and after the seed implantation, and counting the seeds on the postevaluation CT scans. In comparison, examinations relying only on the analysis of postoperative chest radiographs yielded a false-positive detection in four patients and a false negative detection in two patients. Another advantage of the seed-migration detector is that multiple seed-migration scans can be performed without exposing the patient to any additional radiation, for this device is a passive detector. Our clinical implementation also demonstrated that the seed-migration detector is a convenient and cost-effective method. As a result of this study, we stopped ordering the postoperative chest radiographs in a patient's regular postevaluation visit. Only if the detector shows radioactivity outside a patient's pelvis are a pair of anteroposterior and lateral chest radiographs of the patient ordered to document the location of the embolized seeds. PMID- 12772986 TI - Tissue-specific dosimetry for radioiodine therapy of the autonomous thyroid nodule. AB - A tissue-specific dosimetric method based on gamma camera acquisitions was developed to determine the 131I activity to administer to patients with autonomous thyroid nodules (ATN) to deliver 200 Gy to the nodule and to evaluate the correspondent dose to extranodular tissue. Twenty patients with ATN were given 111 MBq of 123I i.v. and their neck was imaged 2, 4, 24, 48, and 120 hours after administration to evaluate separate iodine kinetics for nodule and contralateral lobe. The volumes of nodule and lobe were measured on the 4 hour scintigraphic image, after optimization of the method on a thyroid phantom. Three simplified dosimetric methods were then considered and compared to the reference method in terms of 131I activity: (a) three point method, based on 4, 24, and 120 h acquisitions, (b) fixed T1/2 method, that measures only the 24 h uptake and assumes an effective half-life of 5 days for the nodule, (c) fixed activity method, based on the administration of 413 MBq of 131I. The mean 131I activity to administer to the 20 patients was 413 MBq (range 65-1327) and the mean dose to the contralateral lobe was 43 Gy (range 11-121). The percentage differences in 131I activity between the reference method and the simplified methods were in the ranges: (a) -14%, 13%, (b) -42%, 74%, (c) -69%, 533%. The relevant dose to extranodular tissue and the great interpatient variability of the radioiodine activity required to give a predetermined dose to ATN suggest that a tissue specific dosimetric approach based on gamma camera acquisitions is fundamental. A simple method based on only three uptake measurements is a reliable alternative to the five point method when the clinical workload of a Nuclear Medicine department is particularly heavy. PMID- 12772987 TI - Validation of dynamic MLC-controller log files using a two-dimensional diode array. AB - Intensity-modulated radiation therapy (IMRT) delivered with multi-leaf collimator (MLC) in the step-and-shoot mode uses multiple static MLC segments to achieve intensity modulation. For typical IMRT treatment plans, significant numbers of segments are delivered with monitor units (MUs) of much less than 10. Verification of the ability of the linear accelerator (linac) to deliver small MU segments accurately is an important step in the IMRT commissioning and quality assurance (QA) process. Recent studies have reported large discrepancies between the intended and delivered segment MUs. These discrepancies could potentially cause large errors in the delivered patient dose. We have undertaken a systematic study to evaluate the accuracy of the dynamic MLC log files, which are created automatically by our commercial MLC workstation after each delivery, in recording the fractional MU delivered in the step-and-shoot mode. Two linac models were evaluated with simple-geometry leaf sequences and delivered with different total MUs and different nominal dose rates. A commercial two-dimensional diode array was used for the measurement. Large discrepancies between the intended and delivered segment MUs were found. The discrepancies were larger for small MU segments at higher dose rate, with some small MU segments completely undelivered. The recorded fractional MUs in the log files were found to agree with what was delivered within the limits of our experimental uncertainty. Our results indicate that it is important to verify the delivery accuracy of small MU segments that could potentially occur in a patient treatment and that the log files are useful in checking the integrity of the linac delivery once validated. Thus validated log files can be used as a QA tool for general IMRT delivery and patient-specific plan verification. PMID- 12772988 TI - Analytical linear energy transfer calculations for proton therapy. AB - As the relative biological effectiveness of protons depends on the linear energy transfer (LET), simple methods for LET calculations are desired for the optimization of proton therapy. This work provides an analytical model for the LET on the central axis of broad proton beams in water, which can also be applied to spread-out Bragg peaks. For realistic treatment situations with polyenergetic beams, the LET is here defined as a local mean of the proton stopping power, weighted by the local energy spectrum. The proposed model considers only Coulomb interactions and neglects nonelastic nuclear interactions. By assuming a Gaussian shape for the energy spectrum and by using a suitable parametrization of the stopping power, analytical expressions for the track averaged and the dose averaged LET are derived, which account for range straggling as well as for the initial width of the energy spectrum. The analytical model was evaluated by Monte Carlo simulations with GEANT 3.21. Local energy spectra were simulated to obtain LET distributions for several cases, using clinical energies between 70 and 250 MeV and varying widths of the initial energy spectrum. Good agreement was found between the analytical model and the Monte Carlo simulations (with maximum deviations of 0.5 keV per micrometer), which justifies the assumptions used in the derivation of the analytical model. PMID- 12772989 TI - A pointwise limit theorem for filtered backprojection in computed tomography. AB - Computed tomography (CT) is one of the most important areas in the modern science and technology. The most popular approach for image reconstruction is filtered backprojection. It is essential to understand the limit behavior of the filtered backprojection algorithms. The classic results on the limit of image reconstruction are typically done in the norm sense. In this paper, we use the method of limited bandwidth to handle filtered backprojection-based image reconstruction when the spectrum of an underlying image is not absolutely integrable. Our main contribution is, assuming the method of limited bandwidth, to prove a pointwise limit theorem for a class of functions practically relevant and quite general. Further work is underway to extend the theory and explore its practical applications. PMID- 12772990 TI - The use of a priori information in the detection of mammographic microcalcifications to improve their classification. AB - In this work, we present a calcification-detection scheme that automatically localizes calcifications in a previously detected cluster in order to generate the input for a cluster-classification scheme developed in the past. The calcification-detection scheme makes use of three pieces of a priori information: the location of the center of the cluster, the size of the cluster, and the approximate number of calcifications in the cluster. This information can be obtained either automatically from a cluster-detection scheme or manually by a radiologist. It is used to analyze only the portion of the mammogram that contains a cluster and to identify the individual calcifications more accurately, after enhancing them by means of a "Difference of Gaussians" filter. Classification performances (patient-based Az=0.92; cluster-based Az=0.72) comparable to those obtained by using manually-identified calcifications (patient based Az=0.92; cluster-based Az=0.82) can be achieved. PMID- 12772991 TI - Image reconstruction and image quality evaluation for a 16-slice CT scanner. AB - We present a theoretical overview and a performance evaluation of a novel approximate reconstruction algorithm for cone-beam spiral CT, the adaptive multiple plane reconstruction (AMPR), which has been introduced by Schaller, Flohr et al. [Proc. SPIE Int. Symp. Med. Imag. 4322, 113-127 (2001)] AMPR has been implemented in a recently introduced 16-slice CT scanner. We present a detailed algorithmic description of AMPR which allows for a free selection of the spiral pitch. We show that dose utilization is better than 90% independent of the pitch. We give an overview on the z-reformation functions chosen to allow for a variable selection of the spiral slice width at arbitrary pitch values. To investigate AMPR image quality we present images of anthropomorphic phantoms and initial patient results. We present measurements of spiral slice sensitivity profiles (SSPs) and measurements of the maximum achievable transverse resolution, both in the isocenter and off-center. We discuss the pitch dependence of image noise measured in a centered 20 cm water phantom. Using the AMPR approach, cone beam artifacts are considerably reduced for the 16-slice scanner investigated. Image quality in MPRs is independent of the pitch and equivalent to a single slice CT system at pitch p approximately 1.5. The full width at half-maximum (FWHM) of the spiral SSPs shows only minor variations as a function of the pitch, nominal, and measured values differ by less than 0.2 mm. With 16 x 0.75 mm collimation, the measured FWHM of the smallest reconstructed slice is about 0.9 mm. Using this slice width and overlapping image reconstruction, cylindrical holes with 0.6 mm diameter can be resolved in a z-resolution phantom. Image noise for constant effective mAs is nearly independent of the pitch. Measured and theoretically expected dose utilization are in good agreement. Meanwhile, clinical practice has demonstrated the excellent image quality and the increased diagnostic capability that is obtained with the new generation of multislice CT systems. PMID- 12772992 TI - Dosimetric evaluation of partially overlapping intensity modulated beams using dynamic mini-multileaf collimation. AB - The dose distribution resulting from partially overlapping intensity modulated beams (IMBs) assigned to different isocenters for the treatment of the same planning target volume (PTV) was evaluated. These partially overlapping IMBs are used in static intensity modulated radiation therapy (IMRT) treatments with the Novalis system using the mini-MultiLeaf Collimator (mini-MLC) in Dynamic MultiLeaf Collimation (DMLC) mode. The resultant dose distribution was verified dosimetrically for a cylindrical target defined in a homogeneous cubic phantom. The phantom positioning can introduce dose nonuniformities in the resultant dose distribution by nonperfect positioning of the isocenters in accordance with each other. The dose inhomogeneities are quantified mathematically by summation of the dose profiles of the used IMBs and experimentally by measurement of the resulting dose profiles with radiographic film and thermoluminescent detectors (TLD). The mathematical estimation of the resulting dose profile of the treatment with a perfect positioning of the isocenters showed a good agreement with the planned dose profile. The magnitude of the maximum dose inhomogeneities introduced by the simulated supplementary shifts between the isocenters decreases by -8.54% mm(-1) as the shift changes from -0.30 +/- 0.10 cm to +0.30 +/- 0.10 cm. The TLD measurements showed a similar variation of the magnitude of the maximum dose inhomogeneities: -8.77% mm(-1). The amount of dose variation was underestimated with the radiographic film measurements, which showed a variation of -7.17% mm( 1). The film measurements demonstrated that the magnitude of the introduced maximum dose inhomogeneities did not alter significantly throughout the PTV. The approach of using partially overlapping IMBs assigned to different isocenters to enlarge the treatment region introduces smaller dose inhomogeneities in the resultant dose distribution than when abutting treatment fields are used. The resultant dose distribution of this treatment technique is less sensitive to positioning errors of the used treatment isocenters. PMID- 12772993 TI - Thermoacoustic computed tomography using a conventional linear transducer array. AB - We report on methodology for employing a conventional linear transducer array as a thermoacoustic detector in a thermoacoustic computed tomography (TCT) device, which has been designed for imaging small animals, e.g., athymic nude mice. We tested this concept using a 5 MHz, 128-element linear array (Acuson model L538). Thermoacoustic emissions were induced in a tissue-mimicking phantom using a Nd:YAg laser, operated at 1064 nm. Two-dimensional, axial "slice" images were formed using a filtered-backprojection algorithm. In-plane spatial resolution was measured as better than 200 microns with a slice thickness of 1.5 mm (full width at half maximum). The same detector, when operated as a conventional phased array, produced conventional ultrasound images in perfect registration with the TCT images. PMID- 12772994 TI - Mesh-based enhancement schemes in diffuse optical tomography. AB - Two mesh-based methods including dual meshing and adaptive meshing are developed to improve the finite element-based reconstruction of both absorption and scattering images of heterogeneous turbid media. The idea of dual meshing scheme is to use a fine mesh for the solution of photon propagation and a coarse mesh for the inversion of optical property distributions. The adaptive meshing method is accomplished by the automatic mesh refinement in the region of heterogeneity during reconstruction. These schemes are validated using tissue-like phantom measurements. Our results demonstrate the capabilities of the dual meshing and adaptive meshing in both qualitative and quantitative improvement of optical image reconstruction. PMID- 12772995 TI - A 2-D diode array and analysis software for verification of intensity modulated radiation therapy delivery. AB - An analysis is made of a two-dimensional array of diodes that can be used for measuring dose generated in a plane by a radiation beam. This measuring device is the MapCHECK Model 1175 (Sun Nuclear, Melbourne, FL). This device has 445 N-type diodes in a 22 x 22 cm2 2-D array with variable spacing. The entire array of diodes is easily calibrated to allow for measurements in absolute dose. For IMRT quality assurance, each beam is measured individually with the beam central axis oriented perpendicular to the plane of diodes. Software is available to do the analytical comparison of measurements versus dose distributions calculated by a treatment planning system. Comparison criteria of percent difference and distance to-agreement are defined by the operator. Data are presented that show the diode array has linear response when beam fluence changes by over 300-fold, which is typical of the level of modulation in intensity modulated radiation therapy, IMRT, beams. A linear dependence is also shown for a 100-fold change in monitors units delivered. Methods for how this device can be used in the clinic for quality assurance of IMRT fields are described. Measurements of typical IMRT beams that are modulated by compensators and MLCs are presented with comparisons to treatment planning system dose calculations. A time analysis is done for typical IMRT quality assurance measurements. The setup, calibration, and analysis time for the 2-D diode array are on the order of 20 min, depending on numbers of fields. This is significantly less time than required to do similar analysis with radiographic film. The 2-D diode array is ideal for per-plan quality assurance after an IMRT system is fully commissioned. PMID- 12772996 TI - A multiportal compensator system for IMRT delivery. AB - We have developed a multiportal compensator system for IMRT delivery, comprising a rotational compensator mount for a linac head, cylindrical compensator enclosures positioned in the mount, a vacuum-formed thermoplastic sheet with heavy alloy granules inside the enclosure, and a vacuum thermoforming device. The mount rotates like a revolver by a stepping motor, thus allowing automatic multiportal IMRT without exchanging compensators by human operators during treatment. The thermoforming device has servo-motor-driven 10 x 10 metal rod elements to actualize an arbitrary intensity profile. The thermoplastic sheet is preheated by a built-in biplanar heater and then it is placed over the rod elements. Subsequently, vacuum forming is performed through corner cutouts of the rod elements. After forced cooling down, the heavy alloy granules are fed into the formed sheet. Preliminary experiment using solid water phantoms and an x-ray film has shown that the intensity profile on the film agrees reasonably well with the desired profile. PMID- 12772997 TI - Semiautomatic three-dimensional segmentation of the prostate using two dimensional ultrasound images. AB - In this paper, we report on two methods for semiautomatic three-dimensional (3-D) prostate boundary segmentation using 2-D ultrasound images. For each method, a 3 D ultrasound prostate image was sliced into the series of contiguous 2-D images, either in a parallel manner, with a uniform slice spacing of 1 mm, or in a rotational manner, about an axis approximately through the center of the prostate, with a uniform angular spacing of 5 degrees. The segmentation process was initiated by manually placing four points on the boundary of a selected slice, from which an initial prostate boundary was determined. This initial boundary was refined using the Discrete Dynamic Contour until it fit the actual prostate boundary. The remaining slices were then segmented by iteratively propagating this result to an adjacent slice and repeating the refinement, pausing the process when necessary to manually edit the boundary. The two methods were tested with six 3-D prostate images. The results showed that the parallel and rotational methods had mean editing rates of 20% and 14%, and mean (mean absolute) volume errors of -5.4% (6.5%) and -1.7% (3.1%), respectively. Based on these results, as well as the relative difficulty in editing, we conclude that the rotational segmentation method is superior. PMID- 12772998 TI - Prostate brachytherapy seed identification on post-implant TRUS images. AB - TRUS is a conceptually appealing alternative to CT-based dosimetry, offering the substantial practical advantage of being readily available intraoperatively. To test the feasibility and reliability of seed identification on post-implant TRUS using standard two-dimensional images, ten patients treated with I-125 or Pd-103 brachytherapy were studied. A set of transverse images (6 MHz) were taken immediately following completion of the implant procedure. Original thermal images were sent to four physicians and the sources were identified independently by placing marks on a cellophane overlay, with grids to match the axial TRUS images. The number and type of seed implanted were not revealed to the investigators. Instead, they were instructed to mark the positions of what they would consider, with reasonable certainty, to be seeds. The overlays were then manually compared for source identification and agreement between observers regarding each alleged source. The actual number of implanted seeds ranged from 44 to 108 (median: 60). In contrast, the mean number of seeds allegedly identified per patients ranged from 26 to 82 (median: 43). The average percent of the seeds allegedly identified per patient ranged from 51% to 83% (mean: 74%). The four physician investigators--KW, JS, BH, and GM--identified an alleged median of 90%, 44%, 63%, and 91% of the total seeds, respectively. There were five instances in which investigators alleged more seeds than were actually implanted. The consistency of seed identification among the investigators was evaluated by noting how many investigators identified each bright spot on the images. The percent of bright spots identified by all four investigators ranged from 8% to 33% (median: 20%). Despite considerable interest among some of our clinical and commercial colleagues in developing TRUS-based intraoperative post implant dosimetry, the use of TRUS-based seed identification for post-implant dosimetry should be viewed with skepticism. PMID- 12772999 TI - A submillimeter resolution fluorescence molecular imaging system for small animal imaging. AB - Most current imaging systems developed for tomographic investigations of intact tissues using diffuse photons suffer from a limited number of sources and detectors. In this paper we describe the construction and evaluation of a large dataset, low noise tomographic system for fluorescence imaging in small animals. The system consists of a parallel plate-imaging chamber and a lens coupled CCD camera, which enables conventional planar imaging as well as fluorescence tomography. The planar imaging data are used to guide the acquisition of a Fluorescence Molecular Tomography (FMT) dataset containing more than 106 measurements, and to superimpose anatomical features with tomographic results for improved visual representation. Experimental measurements exhibited good agreement with the diffusion theory models used to predict light propagation within the chamber. Tests of the instrument's capacity to quantitatively reconstruct fluorochrome distributions in three dimensions showed less than 5% errors between actual fluorochrome concentrations and FMT findings, and suggested a detection threshold of approximately 100 femptomoles for small localized objects. Experiments to assess the instrument's spatial resolution demonstrated the ability of the system to resolve objects placed at clear distances of less than 1 mm. This is a significant resolution increase over previously developed systems for animal imaging, and is primarily due to the large dataset employed and the use of inversion methods. Finally, the in vivo imaging capacity is showcased. It is expected that the large dataset collected can enable superior imaging of molecular probes in vivo and improve quantification of fluorescence signatures. PMID- 12773000 TI - Characteristics of sensitometric curves of radiographic films. AB - A new type of radiographic film, EDR (extended dose range) film, has been recently become available for film dosimetry. It is particularly attractive for composite isodose verification of intensity modulated radiation therapy because of its low sensitivity relative to the more common Kodak XV film. For XV film, the relationship between optical density and dose, commonly known as the sensitometric curve, depends linearly on the dose at low densities. Unlike XV film, the sensitometric curve of EDR film irradiated by megavoltage x rays is not linearly dependent on the dose at low densities. In this work, to understand the mechanisms governing the shape of the sensitometric curves, EDR film was studied with kilovoltage x rays, 60Co gamma rays, megavoltage x rays, and electron beams. As a comparison, XV film was also studied with the same beams mentioned above. The model originally developed by Silberstein [J. Opt. Soc. Am. 35, 93-107, 1945)] is used to fit experimental data. It is found that the single hit model can be used to predict the sensitometric curve for XV films irradiated by all beams used in this work and for EDR films exposed to kilovoltage x rays. For EDR film irradiated by 60Co gamma rays, megavoltage x rays, and electron beams, the double hit model is used to fit the sensitometric curves. For doses less than 100 cGy, a systematic difference between measured densities and that predicted by the double hit model is observed. Possible causes of the observed differences are discussed. The results of this work provide a theoretical explanation of the sensitometric behavior of EDR film. PMID- 12773001 TI - Interpretation of pre- versus postimplant TRUS images. AB - In order to summarize the inter-observer variability of pre- and postimplant TRUS image interpretation. Ten patients treated with Pd-103 brachytherapy were studied. Preimplant prostatevolumes ranged from 21 to 51 cm3. The number of sources implanted ranged from 74 to 155, and the number of sources per cm3 prostate volume ranged from 3.0 to 4.3. A set of transverse images (6 MHz) were taken immediately prior to and following source placement. Original printer images were sent to four investigators and the prostate outlined independently on a cellophane overlay. The overlays were digitized into a Varian MMS 7.0 treatment planning system (Charlottesville, VA) for volume determinations. There was moderate interobserver variability in TRUS volume determination, accentuated for the postimplant images. The standard deviations varied from 2% to 13% of the mean (median: 7%) for preimplant volumes, versus 7% to 32% (median: 13%) for postimplant volumes. Interobserver prostatic edge (border) localization variability was greatest at the base and apex, with closer agreement along the posterior border. For preimplant images, the majority of edge points were within 1.0 mm of the mean. At each coordinate, with the exception of the anterior base, the majority of points were within 2.0 mm of the mean. In general, border identification variability was greater in the post implant images. While all prostate imaging modalities suffer from interobserver variability, preimplant and postimplant TRUS appears capable of consistently determining prostatic volume and borders. It appears that intraoperative TRUS-based dosimetry is a practical goal, provided that seed location coordinates can be added to the prostatic edge information derived from TRUS images. PMID- 12773002 TI - Validation of intensity modulation on a commercial treatment planning system. AB - For two years now, a study on intensity modulated radiotherapy (IMRT) has been in progress at the Antoine Lacassagne Hospital Center for Cancer Therapy (in Nice) in collaboration with the University of Nice-Sophia Antipolis. The kind of intensity modulation that was used is the "step and shoot" technique in which the modulated beam is created both by adding andjoining elementary fields. Before carrying out clinical tests, several problems regarding the production of modulated beams has to be mastered. The current developments of our study enable us to dosimetrically produce (in water phantom and in the PMMA phantom) complexmodulated whose segmentation was calculated by one commercial treatment planning system (TPS). Nevertheless, we showed and studied some critical discrepancies between standard clinical calculations and the calculations using field segmentation. We showed that with nonoptimal conditions of segmentation the discrepancies, which are due to the type of algorithm used, could bring about significant errors inside the field of up to 10% of maximum dose. Another point of our study is the quantification and resolution of differences between measurements and calculations due to the internal segmentation of calculated modulated fields and their realization on Linac. Once again, in none optimal conditions of segmentation and inside the field we obtained discrepancies up to 20% of maximum dose between calculations using field segmentation and measurements. That was mainly due to the tongue and groove effect and penumbra phenomena. This study allows us to show that the discrepancies between segmentation calculations and standard clinical calculations should be solved by the use of penumbra models during segmentation calculations. We will introduce both the study and its near-future perspectives. PMID- 12773003 TI - Magnetic resonance imaging-guided attenuation and scatter corrections in three dimensional brain positron emission tomography. AB - Reliable attenuation correction represents an essential component of the long chain of modules required for the reconstruction of artifact-free, quantitative brain positron emission tomography (PET) images. In this work we demonstrate the proof of principle of segmented magnetic resonance imaging (MRI)-guided attenuation and scatter corrections in three-dimensional (3D) brain PET. We have developed a method for attenuation correction based on registered T1-weighted MRI, eliminating the need of an additional transmission (TX) scan. The MR images were realigned to preliminary reconstructions of PET data using an automatic algorithm and then segmented by means of a fuzzy clustering technique which identifies tissues of significantly different density and composition. The voxels belonging to different regions were classified into air, skull, brain tissue and nasal sinuses. These voxels were then assigned theoretical tissue-dependent attenuation coefficients as reported in the ICRU 44 report followed by Gaussian smoothing and addition of a good statistics bed image. The MRI-derived attenuation map was then forward projected to generate attenuation correction factors (ACFs) to be used for correcting the emission (EM) data. The method was evaluated and validated on 10 patient data where TX and MRI brain images were available. Qualitative and quantitative assessment of differences between TX guided and segmented MRI-guided 3D reconstructions were performed by visual assessment and by estimating parameters of clinical interest. The results indicated a small but noticeable improvement in image quality as a consequence of the reduction of noise propagation from TX into EM data. Considering the difficulties associated with preinjection TX-based attenuation correction and the limitations of current calculated attenuation correction, MRI-based attenuation correction in 3D brain PET would likely be the method of choice for the foreseeable future as a second best approach in a busy nuclear medicine center and could be applied to other functional brain imaging modalities such as SPECT. PMID- 12773004 TI - Application of likelihood ratio to classification of mammographic masses; performance comparison to case-based reasoning. AB - The likelihood ratio (LR) is an optimal approach for deciding which of two alternate hypotheses best describes a given situation. We adopted this formalism for predicting whether biopsy results of mammographic masses will be benign or malignant, aiming to reduce the number of biopsies performed on benign lesions. We compared the performance of this LR-based algorithm (LRb) to a case-based reasoning (CBR) classifier, which provides a solution to a new problem using past similiar cases. Each classifier used mammographers' BI-RADS descriptions of mammographic masses as input. The database consisted of 646 biopsy-proven mammography cases. Performance was evaluated using Receiver Operating Characteristic (ROC) analysis, Round Robin sampling, and bootstrap. The ROC areas (AUC) for the LRb and CBR were 0.91+/- 0.01 and 0.92 +/- 0.01, respectively. The partial ROC area index (0.90AUC) was the same for both classifiers, 0.59 +/- 0.05. At a sensitivity of 98%, the CBR would spare 204 (49%) of benign lesions from biopsy; the LRb would spare 209 (51%) benign lesions. The performance of the two classifiers was very similar, with no statistical differences in AUC or 0.90AUC. Although the CBR and LRb originate from different fields of study, their implementations differ only in the estimation of the probability density functions (PDFs) of the feature distributions. The CBR performs this estimation implicitly, while using various similarity metrics. On the other hand, the estimation of the PDFs is specified explicitly in the LRb implementation. This difference in the estimation of the PDFs results in the very small difference in performance, and at 98% sensitivity, both classifiers would spare about half of the benign mammographic masses from biopsy. The CBR and LRb are equivalent methods in implementation and performance. PMID- 12773005 TI - Dosimetry characterization of a 32P source wire used for intravascular brachytherapy with automated stepping. AB - Depth-dose curve measurements and Monte Carlo simulations for a catheter-based 32P intravascular brachytherapy source wire are described. The measured dose rates were obtained using both radiochromic-dye film and an extrapolation chamber (EC). Calibrated radiochromic-dye films were irradiated at distances between 0.5 and 5 mm from the source axis in polystyrene phantoms, and scanned with high resolution densitometers. Measurements with an automated EC with a 1 mm diameter collecting electrode were also performed at a distance of 2 mm from the source in polystyrene. The measured dose rates obtained from the film and EC were divided by the measured source activity to obtain measured values of dose rate per unit contained activity. Dosimetric calculations of the catheter-based 32P wire geometry were also obtained using several Monte Carlo codes (CYLTRAN, MCNP, PENELOPE, and EGS). The measured and calculated values of dose rate per unit contained activity are in good agreement (<10%) within the relevant treatment distances (1 to 4 mm). With carefully selected input parameters, the calculated depth-dose curves using these codes were within 5% at 4 mm depth. At greater depths the discrepancies between the codes increase. We discuss likely mechanisms for these differences. PMID- 12773006 TI - Verification of absolute ionization chamber dosimetry in a proton beam using carbon activation measurements. AB - Reference ionization chamber dosimetry implemented in a clinical proton beam and based on the ICRU 59 recommendations has been verified with an independent carbon activation method. The 12C(p,pn)11C nuclear reaction was used to measure the beam fluence and entrance dose. A method to transfer from the entrance dose to the dose at the ion chamber calibration position has been developed. Measurements performed in a monochromatic 200 MeV beam show that the ratio of absolute doses measured using the carbon activation and the ion chamber methods is 1.017 +/- 0.03 (type A uncertainty). This result is within the uncertainties of both methods employed, which are estimated at +/- 4.3% (carbon activation) and +/- 2.7% (ion chamber calibration). PMID- 12773008 TI - Determination of the 4 mm Gamma Knife helmet relative output factor using a variety of detectors. AB - Though the 4 mm Gamma Knife helmet is used routinely, there is disagreement in the Gamma Knife users community on the value of the 4 mm helmet relative output factor. A range of relative output factors is used, and this variation may impair observations of dose response and optimization of prescribed dose. To study this variation, measurements were performed using the following radiation detectors: silicon diode, diamond detector, radiographic film, radiochromic film, and TLD cubes. To facilitate positioning of the silicon diode and diamond detector, a three-dimensional translation micrometer was used to iteratively determine the position of maximum detector response. Positioning of the films and TLDs was accomplished by manufacturing custom holders for each technique. Results from all five measurement techniques indicate that the 4 mm helmet relative output factor is 0.868 +/- 0.014. Within the experimental uncertainties, this value is in good agreement with results obtained by other investigators using diverse techniques. PMID- 12773007 TI - CERR: a computational environment for radiotherapy research. AB - A software environment is described, called the computational environment for radiotherapy research (CERR, pronounced "sir"). CERR partially addresses four broad needs in treatment planning research: (a) it provides a convenient and powerful software environment to develop and prototype treatment planning concepts, (b) it serves as a software integration environment to combine treatment planning software written in multiple languages (MATLAB, FORTRAN, C/C++, JAVA, etc.), together with treatment plan information (computed tomography scans, outlined structures, dose distributions, digital films, etc.), (c) it provides the ability to extract treatment plans from disparate planning systems using the widely available AAPM/RTOG archiving mechanism, and (d) it provides a convenient and powerful tool for sharing and reproducing treatment planning research results. The functional components currently being distributed, including source code, include: (1) an import program which converts the widely available AAPM/RTOG treatment planning format into a MATLAB cell-array data object, facilitating manipulation; (2) viewers which display axial, coronal, and sagittal computed tomography images, structure contours, digital films, and isodose lines or dose colorwash, (3) a suite of contouring tools to edit and/or create anatomical structures, (4) dose-volume and dose-surface histogram calculation and display tools, and (5) various predefined commands. CERR allows the user to retrieve any AAPM/RTOG key word information about the treatment plan archive. The code is relatively self-describing, because it relies on MATLAB structure field name definitions based on the AAPM/RTOG standard. New structure field names can be added dynamically or permanently. New components of arbitrary data type can be stored and accessed without disturbing system operation. CERR has been applied to aid research in dose-volume-outcome modeling, Monte Carlo dose calculation, and treatment planning optimization. In summary, CERR provides a powerful, convenient, and common framework which allows researchers to use common patient data sets, and compare and share research results. PMID- 12773009 TI - Total body skin imaging as an aid to melanoma detection. AB - Total body skin imaging (TBSI) is being increasingly used as an aid to melanoma detection in high-risk individuals. In this article, we review the rationale, techniques, advantages, and potential pitfalls of TBSI as an aid to melanoma detection. We highlight the technical and clinical considerations relevant to implementation of TBSI in clinical practice. PMID- 12773010 TI - Principles of dermatoscopy of pigmented skin lesions. AB - There has been a dramatic increase in the incidence of malignant melanoma in most parts of the world. Because the tumor thickness is the most important prognostic factor for the prognosis of the malignant melanoma, the early detection of thin melanomas is essential. Dermatoscopy allows the physician to discriminate between melanocytic and nonmelanocytic lesions with high diagnostic accuracy and to detect initial malignant melanomas. We review the principles of dermatoscopy and the differential diagnosis of pigmented skin lesions. Before using the ABCD rule of dermatoscopy to classify melanocytic lesions into benign, suspicious, or malignant, the distinction between melanocytic and nonmelanocytic lesions is necessary. An essential prerequisite for the usefulness of this technique is adequate training. PMID- 12773011 TI - Congenital melanocytic nevi: treatment modalities and management options. AB - Congenital melanocytic nevi can be cosmetically disfiguring, give rise to melanoma, and suggest the presence of neurocutaneous melanocytosis. Management decisions must be tailored for each patient and each nevus, taking into consideration the risk for developing malignancy, risk for developing symptomatic neurocutaneous melanocytosis, cosmetic implications of having the nevus, cosmetic implications of any resultant surgical scars from their removal, adverse effects that the nevus may have on psycho-social development, and the adverse effects and long-term sequelae of any surgical intervention. The advantages and disadvantages of different modalities used in the treatment of congenital melanocytic nevi are discussed. Organizational flow diagrams are presented to help clinicians in managing patients with different sized congenital melanocytic nevi. PMID- 12773012 TI - Current diagnostic problems in melanoma pathology. AB - The histopathological diagnosis of cutaneous melanocytic lesions may be difficult to assess. Frequently encountered diagnostic problems include: 1) Dysplastic nevus or melanoma in situ?; 2) Melanoma in situ or superficial spreading melanoma?; 3) Lentigo maligna or lentigo maligna melanoma?; 4) Compound nevocellular nevus or nevoid melanoma?; and 5) Spitz nevus or Spitzoid melanoma? Moreover, less frequently encountered diagnostic challenges are discussed: 1) Deep penetrating nevus or nodular melanoma?; and 2) Cellular blue nevus or melanoma metastasis? In this contribution, these problems are discussed after a systematic approach involving a concise histopathological description of the classic lesions considered in the differential diagnoses, a presentation of the deviating histopathological features that give rise to the diagnostic problems, and finally diagnostic recommendations on the classification of the problematic lesions. We also briefly discuss the contribution of additional immunohistochemistry and molecular pathology in aiding to establish a correct diagnosis. PMID- 12773013 TI - The new melanoma staging system. AB - The new melanoma staging system from The American Joint Committee on Cancer (AJCC) is described. This major revision includes new criteria for staging the primary tumor (T), metastatic nodes (N) and distant metastases (M) as well as stage groupings. These criteria more accurately reflect those prognostic features of the primary and metastatic melanoma that correlate with survival outcome. Physicians managing melanoma should use this staging system in their clinical practice and in the conduct of melanoma clinical trials. PMID- 12773014 TI - Adjuvant therapy of melanoma. PMID- 12773015 TI - Melanoma vaccines: early progress and future promises. AB - Melanoma vaccines aim to stimulate host immune responses against the patient's own tumor. A large number of immunotherapeutic interventions have been already studied in small-scale phase I-II trials. These approaches have been based upon presumptions of investigators, including the format in which antigen would be best administered (peptide, protein, tumor lysate, whole tumor cells or genetic material coding for the proteins of interest), the antigen delivery system and adjuvants (cytokines and dendritic cells, gene-therapy), and the route/schedule of administration. Several approaches have already demonstrated an impact on the immune system but very rarely to date upon the patients' clinical disease outcome. Recent developments in cancer immunology have helped elucidate the role of the main players in the development of host anti-tumor immune responses (including the tumor cells, different T cell subsets, and dendritic cells). These research efforts have provided the basis for the multiple vaccine trial interventions that have been proposed to boost host anti-tumor immune responses. They also have allowed the development of multiple new tools to monitor immune responses in vaccinated patients (including ELISPOT, tetramers and intracellular cytokine-release assays, as well as real-time-PCR and analysis of the T cell receptor). Although multiple cancer vaccine approaches have successfully stimulated T cell immune responses in patients with cancer, the most promising ones need to be formally tested for their ability to improve both the immunological and clinical status of the vaccinated patients in phase III randomized clinical trials. PMID- 12773016 TI - Study samples are too small to produce sufficiently precise reliability coefficients. AB - In a survey of journal articles, test manuals, and test critique books, the author found that a mean sample size (N) of 260 participants had been used for reliability studies on 742 tests. The distribution was skewed because the median sample size for the total sample was only 90. The median sample sizes for the internal consistency, retest, and interjudge reliabilities were 182, 64, and 36, respectively. The author presented sample size statistics for the various internal consistency methods and types of tests. In general, the author found that the sample sizes that were used in the internal consistency studies were too small to produce sufficiently precise reliability coefficients, which in turn could cause imprecise estimates of examinee true-score confidence intervals. The results also suggest that larger sample sizes have been used in the last decade compared with those that were used in earlier decades. PMID- 12773017 TI - The S-S construct of expectancy versus the S-R construct of fear: which motivates the acquisition of avoidance behavior? AB - The authors provided a differential test between stimulus-stimulus (S-S) and stimulus-response (S-R) theory predictions in regard to the roles that the constructs of expectancy and of fear play in maintaining classically conditioned fear responding within the context of a human conditioned-avoidance paradigm. After the participants had developed sustained avoidance responding, their shock electrodes and avoidance response apparatus were removed to enhance the cognitive expectancy that the conditioned stimulus (CS) would not be followed by the unconditioned stimulus (UCS). This manipulation of expectancy was successful in 96% of the participants. The study was conducted over a 2-day period and involved 1 experimental group and 3 control groups. During the test trials, the authors used autonomic and self-report indices of fear to assess the presence or absence of fear to the CS. The data disconfirmed the prediction of the S-S theory that fear to the CS would be extinguished. The authors discuss the implications of this finding for S-S theories and for approaches in cognitive behavior therapy. PMID- 12773018 TI - A Monte Carlo evaluation of tests for comparing dependent correlations. AB - The authors conducted a Monte Carlo simulation of 8 statistical tests for comparing dependent zero-order correlations. In particular, they evaluated the Type I error rates and power of a number of test statistics for sample sizes (Ns) of 20, 50, 100, and 300 under 3 different population distributions (normal, uniform, and exponential). For the Type I error rate analyses, the authors evaluated 3 different magnitudes of the predictor-criterion correlations (rho(y,x1) = rho(y,x2) = .1, .4, and .7). For the power analyses, they examined 3 different effect sizes or magnitudes of discrepancy between rho(y,x1) and rho(y,x2) (values of .1, .3, and .6). They conducted all of the simulations at 3 different levels of predictor intercorrelation (rho(x1,x2) = .1, .3, and .6). The results indicated that both Type I error rate and power depend not only on sample size and population distribution, but also on (a) the predictor intercorrelation and (b) the effect size (for power) or the magnitude of the predictor-criterion correlations (for Type I error rate). When the authors considered Type I error rate and power simultaneously, the findings suggested that O. J. Dunn and V. A. Clark's (1969) z and E. J. Williams's (1959) t have the best overall statistical properties. The findings extend and refine previous simulation research and as such, should have greater utility for applied researchers. PMID- 12773019 TI - An examination of the value of the generation effect for learning new material. AB - The generation effect involves an improvement in memory when learners must complete or modify materials. Several researchers have suggested that this effect involves enhanced access to learners' existing memory representations; therefore, the effect should be less effective with meaningless, low meaningful, or unfamiliar material. In the present study, the authors conducted 4 experiments in which legal nonwords were used, and they found no generation effect. In another 2 experiments in which familiar cliches were contrasted with new sentences and with unfamiliar sentences from textbooks, the results showed a greatly reduced generation effect for the new, unfamiliar material. Those findings suggest that memory strategies that depend on the generation effect will have limited effectiveness when they are applied to new or unfamiliar material. PMID- 12773020 TI - Thought suppression in spider-fearful and nonfearful individuals. AB - The authors examined the suppression of spider-related thoughts in spider-fearful (n = 23) and nonfearful (n = 22) individuals. Participants were primed with vivid pictures of spiders and a story about spiders. Next, they were randomly assigned to 1 of 2 conditions: (a) suppression of thoughts associated with the previously presented spider-related stimuli or (b) free expression of any thoughts, including those related to the spider-related stimuli. All participants completed a subsequent free-expression exercise. Results indicated that spider-fearful individuals expressed thoughts about the spider-related stimuli for a longer length of time than did nonfearful individuals, particularly in the suppression condition. Participants in both groups demonstrated a rebound of thoughts associated with the spider-related stimuli following suppression. The authors propose that the priming of feared stimuli makes suppression of fear-related thoughts particularly difficult for fearful individuals, perhaps by activating a state of heightened arousal. PMID- 12773021 TI - Coloring only a single letter does not eliminate color-word interference in a vocal-response Stroop task: automaticity revealed. AB - The presence of an interference effect in naming the print color of color words (J. R. Stroop, 1935) suggests that responses associated with the irrelevant-word dimension of the display are activated involuntarily. In the present study, the author examined the conditions under which coloring a single letter in a word reduced interference in vocal responding (D. Kahneman & A. Henik, 1981) or eliminated it in manual responding (D. Besner, J. A. Stolz, & C. Boutilier, 1997). In Experiment 1, color-word interference was significant under vocal responding for the Besner et al. displays. In Experiment 2, the author replicated the Kahneman and Henik effect with the Besner et al. stimuli. The results of Experiment 3 showed that semantic effects are not eliminated by coloring only a single letter. Coloring a single letter does not prevent the activation of the irrelevant-word dimension of the colored color word. PMID- 12773022 TI - Attitude and peer influences on adolescent substance use: the moderating effect of age, sex, and substance. AB - Many studies have suggested the importance of peer influence and personal attitudes (e.g., expectancies, resistance self-efficacy, and perceived harm) in predicting adolescent use of illegal substances. The present study examined these variables in relation to self-reported use of alcohol, cigarettes, and marijuana for 213 younger adolescents (12-15 years old) and 219 older adolescents (18-22 years old). A series of logistic regressions was performed to assess variables relating to use of each substance by age group and gender. Friends' use was significantly related to substance use for both age groups, both sexes, and all substances examined in this study. Perceived harm was not significantly related to use for any group. Finally, outcome expectancies and resistance self-efficacy were differentially related to use depending on age, gender, and substance. The implications of these findings for prevention programming and future research are also discussed. PMID- 12773023 TI - Predictors of substance abuse treatment outcomes in Tennessee. AB - In planning and implementing programs to treat substance abuse, it is important to understand which factors influence post-treatment abstinence. This article identifies and analyzes several variables important in predicting the likelihood of abstinence among substance abuse clients. The data used in this study was collected from 1,350 clients treated for alcohol or drug abuse in residential, halfway house, or outpatient facilities in Tennessee. We analyzed 22 variables as possible treatment outcome predictors by using two statistical procedures: stepwise logistic regression analysis and Quick, Unbiased, Efficient, Statistical Tree (QUEST) analysis, a tree-structured classification algorithm analysis. We found one pretreatment, five in-treatment, and three post-treatment variables to be significant predictors of treatment outcome: previous treatment history, perceived helpfulness of the treatment, simultaneous treatment for mental health, number of days in treatment,completion of treatment, special skills training during treatment, obtaining healthcare services for major physical health problem after treatment, living with someone using alcohol or drugs post treatment, and arrest record since treatment. PMID- 12773024 TI - The relationship between sexual abuse and drug use: findings from Houston's Safer Choices 2 program. AB - Self-report drug use data were collected from 282 female alternative school students surveyed through the Safer Choices 2 study in Houston, Texas. Data collection took place between October 2000 and March 2001 via audio-enabled laptop computers equipped with headphones. Logistic regression analyses indicated that sexual abuse history was significantly associated with lifetime use (OR = 1.9, p < or = 0.05). While the relationships tested in this study are exploratory, they provide evidence for an important connection between sexual abuse and substance use among female alternative school students. PMID- 12773025 TI - Correlates of ecstasy use among students surveyed through the 1997 College Alcohol Study. AB - Anecdotal reports have suggested that the use of 3,4 methylenedioxymethamphetamine (MDMA or "ecstasy") is a growing problem across the United States, primarily among college students and rave attendees. To assess this contention, the drug-using behaviors of 14,520 college students were examined with data collected through the 1997 College Alcohol Study (CAS). Prevalence estimates of ecstasy use were generated and associations between ecstasy use, demographic characteristics, and alcohol and other drug (AOD) use were explored. Six percent of the sample reported lifetime ecstasy use, 3 percent reported use within the past 12 months, and 1 percent reported use within the past 30 days. Compared to non-users, 12-month ecstasy users were significantly more likely to be white, to be a member of a fraternity/sorority, and to have used all other drugs of abuse during the past 12 months. Implications for these findings are discussed. PMID- 12773026 TI - Attitudinal and normative predictors of alcohol use by older adolescents and young adults. AB - A model of alcohol use based on the theory of planned behavior, expectancy theory, and the developmental literature on the influence of parents and peers was examined with 87 eleventh grade students, 105 college freshmen, and 107 college juniors. Specifically, the influence of attitudes about the positive and negative consequences of drinking, perceived parental and peer norms about alcohol consumption, and perceived control over drinking predicted self-reported alcohol use. The results suggest that, during adolescence, decisions to consume alcohol are rational, based on the consideration of the positive consequences of alcohol use and perceptions of control over drinking; however, the negative consequences of alcohol use are discounted. While perceived peer norms predicted alcohol consumption in all three age groups, the influence of perceived parental norms varied such that they predicted alcohol use only among the college juniors. Implications and avenues for future research are discussed. PMID- 12773027 TI - Risk, protection, and substance use in adolescents: a multi-site model. AB - This article reports findings from a national longitudinal cross-site evaluation of high-risk youth to clarify the relationships between risk and protective factors and substance use. Using structural equation modeling, baseline data on 10,473 youth between the ages of 9 and 18 in 48 high-risk communities around the nation are analyzed. Youth were assessed on substance use (cigarette, alcohol, and marijuana use), external risk factors including family, school, peer and neighborhood influences, and individual risk and protective factors including self-control, family connectedness, and school connectedness. Findings indicate strong direct relationships between peer and parental substance use norms and substance use. Individual protective factors, particularly family and school connectedness were strong mediators of individual substance use. These findings suggest that multi-dimensional prevention programming stressing the fostering of conventional anti-substance use attitudes among parents and peers, the importance of parental supervision, and development of strong connections between youth and their family, peers, and school may be most effective in preventing and reducing substance use patterns among high-risk youth. PMID- 12773028 TI - Provision of hepatitis C education in a nationwide sample of drug treatment programs. AB - Hepatitis C virus (HCV) has reached epidemic proportions among drug users, and drug programs are in a unique position to provide each of their patients with HCV education. Using a nationwide sample (N = 434) of drug treatment programs, we report the results of a logistic regression analysis that differentiates programs providing HCV education to all of their patients versus programs that do not. Fifty-four percent of the programs provide HCV education to all of their patients. Programs are about four and a half times as likely to provide HCV education to all patients if they dispense methadone; almost four times as likely to provide this service if they educate most of their staff about HCV; twice as likely if they are residential; and almost twice as likely if they conduct HIV testing on-site. Our findings indicate that there is a need to increase HCV educational services in drug treatment programs. PMID- 12773029 TI - Angiotensin II AT1 receptor antagonists. Clinical implications of active metabolites. PMID- 12773030 TI - Isoform-selective substrates of nitric oxide synthase. AB - Because of the double-edged nature of NO, the development of isoform-selective NOS substrates is a highly desirable goal. Given the striking similarity in the heme active sites of the three NOS isoforms, it presents an challenging problem. Several N-aryl-N'-hydroxyguanidines have recently been shown as substrates that are selective for iNOS over nNOS. Here, we report the first success that 3 is a good substrate for nNOS (70% activity of NOHA, K(m) approximately 40 +/- 6 microM) over iNOS. PMID- 12773031 TI - Design and synthesis of highly potent benzodiazepine gamma-secretase inhibitors: preparation of (2S,3R)-3-(3,4-difluorophenyl)-2-(4-fluorophenyl)-4- hydroxy-N ((3S)-1-methyl-2-oxo-5- phenyl-2,3-dihydro-1H-benzo[e][1,4]-diazepin-3 yl)butyramide by use of an asymmetric Ireland-Claisen rearrangement. AB - Novel benzodiazepine-containing gamma-secretase inhibitors for potential use in Alzheimer's disease have been designed that incorporate a substituted hydrocinnamide C-3 side chain. A syn combination of alpha-alkyl or aryl and beta hydroxy or hydroxymethyl substituents was shown to give highly potent compounds. In particular, (2S,3R)-3-(3,4-difluorophenyl)-2-(4-fluorophenyl)-4-hydroxy-N ((3S)-2-oxo-5-phenyl-2,3-dihydro-1H-benzo[e][1,4]diazepin-3-yl)butyramide (34) demonstrated excellent in vitro potency (IC(50) = 0.06 nM). 34 could also be selectively methylated to give [(3)H]-28, which is of use in radioligand binding assays. PMID- 12773032 TI - 3-(4-[[Benzyl(methyl)amino]methyl]phenyl)-6,7-dimethoxy-2H-2-chromenone (AP2238) inhibits both acetylcholinesterase and acetylcholinesterase-induced beta-amyloid aggregation: a dual function lead for Alzheimer's disease therapy. AB - In recent years, the investigation of acetylcholinesterase (AChE) inhibitors has gained further interest, because the involvement of the peripheral site of the enzyme in the beta-amyloid (Abeta) aggregation process has been disclosed. We present here, for the first time, a direct evidence of the Abeta antiaggregating action of an AChE inhibitor (AP2238) purposely designed to bind at both the catalytic and the peripheral sites of the human enzyme. PMID- 12773033 TI - A highly selective, non-hydantoin, non-carboxylic acid inhibitor of aldose reductase with potent oral activity in diabetic rat models: 6-(5-chloro-3 methylbenzofuran- 2-sulfonyl)-2-H-pyridazin-3-one. AB - We report here on the discovery path that led to a structurally unprecedented non hydantoin, non-carboxylic acid aldose reductase inhibitor, 24, which shows remarkably potent oral activity in normalizing elevated sorbitol levels and, more significantly, fructose levels in the sciatic nerve of chronically diabetic rats, with ED(90) values of 0.8 and 3 mpk, respectively. It is well absorbed in rats (oral bioavailability, 98%) and has a long plasma t(1/2) (26 +/- 3 h). PMID- 12773034 TI - Comparative evaluation of 11 scoring functions for molecular docking. AB - Eleven popular scoring functions have been tested on 100 protein-ligand complexes to evaluate their abilities to reproduce experimentally determined structures and binding affinities. They include four scoring functions implemented in the LigFit module in Cerius2 (LigScore, PLP, PMF, and LUDI), four scoring functions implemented in the CScore module in SYBYL (F-Score, G-Score, D-Score, and ChemScore), the scoring function implemented in the AutoDock program, and two stand-alone scoring functions (DrugScore and X-Score). These scoring functions are not tested in the context of a particular docking program. Instead, conformational sampling and scoring are separated into two consecutive steps. First, an exhaustive conformational sampling is performed by using the AutoDock program to generate an ensemble of docked conformations for each ligand molecule. This conformational ensemble is required to cover the entire conformational space as much as possible rather than to focus on a few energy minima. Then, each scoring function is applied to score this conformational ensemble to see if it can identify the experimentally observed conformation from all of the other decoys. Among all of the scoring functions under test, six of them, i.e., PLP, F Score, LigScore, DrugScore, LUDI, and X-Score, yield success rates higher than the AutoDock scoring function. The success rates of these six scoring functions range from 66% to 76% if using root-mean-square deviation < or =2.0 A as the criterion. Combining any two or three of these six scoring functions into a consensus scoring scheme further improves the success rate to nearly 80% or even higher. However, when applied to reproduce the experimentally determined binding affinities of the 100 protein-ligand complexes, only X-Score, PLP, DrugScore, and G-Score are able to give correlation coefficients over 0.50. All of the 11 scoring functions are further inspected by their abilities to construct a descriptive, funnel-shaped energy surface for protein-ligand complexation. The results indicate that X-Score and DrugScore perform better than the other ones at this aspect. PMID- 12773035 TI - Novel dihydrofolate reductase inhibitors. Structure-based versus diversity-based library design and high-throughput synthesis and screening. AB - Novel 2,4-diaminopyrimidines bearing N,N-disubstituted aminomethyl residues at the 5-position were designed as dihydrofolate reductase (DHFR) inhibitors. These compounds were obtained by treatment of 1-[(2,4-diamino-5 pyrimidinyl)methyl]pyridinium bromide with secondary amines in a polar solvent and in the presence of triethylamine at room temperature. The procedure was found to be very efficient and suitable for application in high-throughput synthesis. In addition, we found that high-throughput screening for enzymatic and in vitro antibacterial activity could be performed on crude reaction mixtures, thus avoiding any purification step. Over 1200 proprietary secondary amines were selected for high-throughput synthesis, based on structural and diversity-related criteria, and the resulting products were submitted to high-throughput screening. A greater number of hits, and significantly more active compounds, were obtained through structure-based library design than through diversity-based library design. Different classes of inhibitors of DHFR were identified in this way, including compounds derived from di-, tri-, and tetracyclic amines. In general, these products showed high activity against the enzymes derived from both TMP sensitive and TMP-resistant Streptococcus pneumoniae. Some compounds possessed appreciable selectivity for the bacterial over the human enzyme, whereas other compounds were not at all selective. In most cases, active enzyme inhibitors also displayed antibacterial activity. PMID- 12773036 TI - Predicting drug metabolism: a site of metabolism prediction tool applied to the cytochrome P450 2C9. AB - The aim of the present study is to develop a method for predicting the site at which molecules will be metabolized by CYP 2C9 (cytochrome P450 2C9) using a previously reported protein homology model of the enzyme. Such a method would be of great help in designing new compounds with a better pharmacokinetic profile, or in designing prodrugs where the compound needs to be metabolized in order to become active. The methodology is based on a comparison between alignment independent descriptors derived from GRID Molecular Interaction Fields for the CYP 2C9 active site, and a distance-based representation of the substrate. The predicted site of metabolism is reported as a ranking list of all the hydrogen atoms of each substrate molecule. Eighty-seven CYP 2C9-catalyzed oxidative reactions reported in the literature have been analyzed. In more than 90% of these cases, the hydrogen atom ranked at the first, second, or third position was the experimentally reported site of oxidation. PMID- 12773037 TI - Introduction of a chemical constraint in a short peptide derived from human acidic fibroblast growth factor elicits mitogenic structural determinants. AB - Fibroblast growth factors (FGFs) are regulatory proteins associated with a number of physiological and pathological states. On the basis of data suggesting a functional role for specific regions of human acidic FGF (aFGF), a linear peptide encompassing residues 99-108 (peptide1) and its cyclic analogue (peptide 2) were synthesized and their functional and structural features were investigated. While peptide 1 is inactive on Balb/c 3T3 fibroblasts, peptide 2 is mitogenic with ED(50) of approximately 50 microM. Moreover, peptide 1 is not able to inhibit the binding of human aFGF to cellular receptors whereas peptide 2 exhibits significant inhibitory activity. The NMR-derived solution conformers indicated the presence, only in peptide 2, of structural elements that we believe are related to its ability to emulate the biological activity of the native protein. These results suggest that the expression of mitogenic activity in short peptides, besides the presence of specific amino acids, requires the existence of stable structural features. In addition, they indicate that the introduction of chemical restraints in peptides can provide novel possibilities for the development of receptor agonists or antagonists. PMID- 12773038 TI - A novel somatostatin mimic with broad somatotropin release inhibitory factor receptor binding and superior therapeutic potential. AB - A rational drug design approach, capitalizing on structure-activity relationships and involving transposition of functional groups from somatotropin release inhibitory factor (SRIF) into a reduced size cyclohexapeptide template, has led to the discovery of SOM230 (25), a novel, stable cyclohexapeptide somatostatin mimic that exhibits unique high-affinity binding to human somatostatin receptors (subtypes sst1-sst5). SOM230 has potent, long-lasting inhibitory effects on growth hormone and insulin-like growth factor-1 release and is a promising development candidate currently under evaluation in phase I clinical trials. PMID- 12773039 TI - Three dimensional pharmacophore modeling of human CYP17 inhibitors. Potential agents for prostate cancer therapy. AB - We report here a molecular modeling investigation of steroidal and nonsteroidal inhibitors of human cytochrome P450 17alpha-hydroxylase-17,20-lyase (CYP17). Using the pharmacophore perception technique, we have generated common-feature pharmacophore model(s) to explain the putative binding requirements for two classes of human CYP17 inhibitors. Common chemical features in the steroid and nonsteroid human CYP17 enzyme inhibitors, as deduced by the Catalyst/HipHop program, are one to two hydrogen bond acceptors (HBAs) and three hydrophobic groups. For azole-steroidal ligands, the 3beta-OH group of ring A and the N-3 of the azole ring attached to ring D at C-17 act as hydrogen bond acceptors. A model that permits hydrogen bond interaction between the azole functionality on ring D and the enzyme is consistent with experimental deductions for type II CYP17 inhibitors where a sixth ligating atom interacts with Fe(II) of heme. In general, pharmacophore models derived for steroid and nonsteroidal compounds bear striking similarities to all azole sites mapping the HBA functionality and to three hydrophobic features describing the hydrophobic interactions between the ligands and the enzyme. Using the pharmacophore model derived for azole-steroidal inhibitors as a 3D search query against several 3D multiconformational Catalyst formatted databases, we identified several steroidal compounds with potential inhibition of this enzyme. Biological testing of some of these compounds show low to high inhibitory potency against the human CYP17 enzyme. This shows the potential of our pharmacophore model in identifying new and potent CYP17 inhibitors. Further refinement of the model is in progress with a view to identifying and optimizing new leads. PMID- 12773040 TI - Design, synthesis, and structure-activity relationships of alkylcarbamic acid aryl esters, a new class of fatty acid amide hydrolase inhibitors. AB - Fatty acid amide hydrolase (FAAH), an intracellular serine hydrolase enzyme, participates in the deactivation of fatty acid ethanolamides such as the endogenous cannabinoid anandamide, the intestinal satiety factor oleoylethanolamide, and the peripheral analgesic and anti-inflammatory factor palmitoylethanolamide. In the present study, we report on the design, synthesis, and structure-activity relationships (SAR) of a novel class of potent, selective, and systemically active inhibitors of FAAH activity, which we have recently shown to exert potent anxiolytic-like effects in rats. These compounds are characterized by a carbamic template substituted with alkyl or aryl groups at their O- and N-termini. Most compounds inhibit FAAH, but not several other serine hydrolases, with potencies that depend on the size and shape of the substituents. Initial SAR investigations suggested that the requirements for optimal potency are a lipophilic N-alkyl substituent (such as n-butyl or cyclohexyl) and a bent O aryl substituent. Furthermore, the carbamic group is essential for activity. A 3D QSAR analysis on the alkylcarbamic acid aryl esters showed that the size and shape of the O-aryl moiety are correlated with FAAH inhibitory potency. A CoMSIA model was constructed, indicating that whereas the steric occupation of an area corresponding to the meta position of an O-phenyl ring improves potency, a region of low steric tolerance on the enzyme active site exists corresponding to the para position of the same ring. The bent shape of the O-aryl moieties that best fit the enzyme surface closely resembles the folded conformations observed in the complexes of unsaturated fatty acids with different proteins. URB524 (N cyclohexylcarbamic acid biphenyl-3-yl ester, 9g) is the most potent compound of the series (IC(50) = 63 nM) and was therefore selected for further optimization. PMID- 12773041 TI - Synthesis and structure-activity relationship of alpha-sulfonylhydroxamic acids as novel, orally active matrix metalloproteinase inhibitors for the treatment of osteoarthritis. AB - The matrix metalloproteinases (MMPs) are a family of zinc-containing endopeptidases that play a key role in both physiological and pathological tissue degradation. These enzymes are strictly regulated by endogenous inhibitors such as tissue inhibitors of MMPs and alpha(2)-macroglobulins. Overexpression of these enzymes has been implicated in various pathological disorders such as arthritis, tumor metastasis, cardiovascular diseases, and multiple sclerosis. Developing effective small-molecule inhibitors to modulate MMP activity is one approach to treat these degenerative diseases. The present work focuses on the discovery and SAR of novel N-hydroxy-alpha-phenylsulfonylacetamide derivatives, which are potent, selective, and orally active MMP inhibitors. PMID- 12773042 TI - Synthesis and structure-activity relationship of N-substituted 4 arylsulfonylpiperidine-4-hydroxamic acids as novel, orally active matrix metalloproteinase inhibitors for the treatment of osteoarthritis. AB - The matrix metalloproteinases (MMPs) are a family of zinc-containing endopeptidases that play a key role in both physiological and pathological tissue degradation. In our preceding paper, we have reported on a series of novel and orally active N-hydroxy-alpha-phenylsulfonylacetamide derivatives. However, these compounds had two drawbacks (moderate selectivity and chirality issues). To circumvent these two problems, a series of novel and orally active N-substituted 4-benzenesulfonylpiperidine-4-carboxylic acid hydroxyamide derivatives have been synthesized. The present paper deals with the synthesis and SAR of these compounds. Among the several compounds synthesized, derivative 55 turned out to be a potent, selective, and an orally active MMP inhibitor in the clinically relevant advanced rabbit osteoarthritis model. Detailed pharmacokinetics and metabolism data are described. PMID- 12773043 TI - Synthesis of ethyl 5-phenyl-6-oxa-1-azabicyclo[3.1.0]hexane-2-carboxylate derivatives and evaluation of their antimalarial activities. AB - Derivatives of ethyl 5-phenyl-6-oxa-1-azabicyclo[3.1.0]hexane-2-carboxylate (14 20), with side chains varying from three to five carbon atoms and bearing various substituents, have been prepared from ethyl 2-phenyl-1-pyrroline-5-carboxylate (12). Their in vitro activity against P. falciparum (K1 strain) and antimycobacterium and also their cytotoxic activity against Vero cell have been evaluated. PMID- 12773044 TI - Benzoylalanine-derived ketoamides carrying vinylbenzyl amino residues: discovery of potent water-soluble calpain inhibitors with oral bioavailability. AB - Novel benzoylalanine-derived ketoamides were prepared and evaluated for calpain I inhibition. Derivatives carrying vinylbenzyl amino residues in the P(2)-P(3) region inhibited calpain in nanomolar concentrations and thus represent a novel class of nonpeptidic calpain inhibitors. Selected examples exhibited an improved pharmacokinetic profile including improved water-solubility and metabolic stability. In particular, these calpain inhibitors showed oral bioavailability in rats as demonstrated by N-(1-benzyl-2-carbamoyl-2-oxoethyl)-2-[E-2-(4 diethylaminomethylphenyl)ethen-1-yl]benzamide (5d). The closely related derivative N-(1-carbamoyl-1-oxohex-1-yl)-2-[E-2-(4-dimethylaminomethylphenyl) ethen-1-yl]benzamide (5b) was evaluated for neuroprotective efficacy after experimental traumatic brain injury in a fluid percussion model in rats. When administered after injury, 5b reduced the number of damaged neurons by 41%, and this result would be in line with the suggested neuroprotective efficacy of calpain inhibition. PMID- 12773045 TI - Optimization of a tertiary alcohol series of phosphodiesterase-4 (PDE4) inhibitors: structure-activity relationship related to PDE4 inhibition and human ether-a-go-go related gene potassium channel binding affinity. AB - A SAR study on the tertiary alcohol series of phosphodiesterase-4 (PDE4) inhibitors related to 1 is described. In addition to inhibitory potency against PDE4 and the lipopolysaccharide-induced production of TNFalpha in human whole blood, the binding affinity of these compounds for the human ether-a-go-go related gene (hERG) potassium channel (an in vitro measure for the potential to cause QTc prolongation) was assessed. Four key structural moieties in the molecule were studied, and the impact of the resulting modifications in modulating these activities was evaluated. From these studies, (+)-3d (L-869,298) was identified as an optimized structure with respect to PDE4 inhibitory potency, lack of binding affinity to the hERG potassium channel, and pharmacokinetic behavior. (+)-3d exhibited good in vivo efficacy in several models of pulmonary function with a wide therapeutic index with respect to emesis and prolongation of the QTc interval. PMID- 12773046 TI - Modifications to the N-terminus but not the C-terminus of calcitonin gene-related peptide(8-37) produce antagonists with increased affinity. AB - Seventeen novel analogues of human calcitonin gene-related peptide(8-37) (hCGRP(8 37)) were synthesized by solid-phase methods and purified to apparent homogeneity by semipreparative cation exchange and/or reversed-phase high-performance liquid chromatography. The C-terminal Phe was replaced by Gly, cyclohexylalanine (Cha), Tyr, all four isomers of beta-methylphenylalanine (beta-MePhe), and l- and d tetrahydroisoquinoline carboxylic acid (Tic), resulting in analogues 3-11. For the synthesis of the beta-MePhe-containing analogues 6-9, crystallization was used to separate a mixture of all four isomers of beta-MePhe into the erythro pair of enantiomers (2S,3S, 2R,3R) and the threo pair of enantiomers (2S,3R, 2R,3S), which were then converted to Fmoc derivatives and used in two separate syntheses. Two diastereomeric peptides were obtained from each synthesis and were separated by RP-HPLC to yield enantiomerically pure 6-9. Substitution of Tyr for Phe caused no change in binding affinity at CGRP receptors. All other substitutions for Phe resulted in substantial reductions in binding affinity. Indeed, no binding was observed for analogues 7, 9, and 11, all of which contained a d-amino acid residue in the C-terminal position, and the binding affinities of the remaining analogues were >10-fold lower than that of h-alpha CGRP(8-37). These data suggest that a conformationally flexible phenyl ring in the C-terminal position of h-alpha-CGRP(8-37) is preferred for high-affinity binding to CGRP receptors. Acetylation, benzoylation, and benzylation of the N termini of h-alpha-CGRP(8-37) and h-beta-CGRP(8-37) produced analogues 12-14 and 16-18, respectively. A byproduct was isolated by RP-HPLC from the resin-cleaved crude product of each benzylated analogue, which was characterized as the dibenzylated derivative of h-alpha-CGRP(8-37) and h-beta-CGRP(8-37) (analogues 15 and 19, respectively). Amino acid analysis and (1)H NMR showed that the second benzyl group was located on the C4 carbon of the imidazole ring of His(10). Radioligand binding experiments showed that derivatizing the N-termini substantially increased binding affinities at CGRP receptors. The benzoylated and dibenzylated derivatives had the highest affinities, which were approximately 50 fold greater than those of h-alpha-CGRP(8-37). Functional experiments confirmed that the N-terminally derivatized analogues of h-alpha-CGRP(8-37) are antagonists that are more potent than h-alpha-CGRP(8-37). In conclusion, these studies underscore the importance of Phe(37) of h-alpha-CGRP(8-37) for binding to CGRP receptors and have identified the N-terminus and His(10) as two positions that can be used for the design of antagonists with increased affinity for CGRP receptors. PMID- 12773047 TI - Synthesis and biological activity of various derivatives of a novel class of potent, selective, and orally active prostaglandin D2 receptor antagonists. 1. Bicyclo[2.2.1]heptane derivatives. AB - Novel prostaglandin D(2) (PGD(2)) receptor antagonists were synthesized as a potential new class of antiallergic agents having a bicyclo[2.2.1]heptane ring system with sulfonamide groups. Some of them exhibit extremely potent antagonism of the PGD(2) receptor in radioligand binding and cAMP formation assays with IC(50) values below 50 nM and much less antagonism of TXA(2) and PGI(2) receptors. These potent PGD(2) receptor antagonists, when given orally, dramatically suppress various allergic inflammatory responses such as increased vascular permeability in allergic rhinitis, conjunctivitis, and asthma models. The excellent pharmacological profiles of PGD(2) receptor antagonists, originally synthesized in our laboratories, are of potentially great clinical significance. This study also provides experimental evidence suggesting that PGD(2) plays an important role in the pathogenesis of allergic diseases. PMID- 12773048 TI - Synthesis and biological activity of various derivatives of a novel class of potent, selective, and orally active prostaglandin D2 receptor antagonists. 2. 6,6-Dimethylbicyclo[3.1.1]heptane derivatives. AB - In an earlier paper, we reported that novel prostaglandin D(2) (PGD(2)) receptor antagonists having the bicyclo[2.2.1]heptane ring system as a prostaglandin skeleton were a potent new class of antiallergic agents and suppressed various allergic inflammatory responses such as those observed in conjunctivitis and asthma models. In the present study, we synthesized PGD(2) receptor antagonists having the 6,6-dimethylbicyclo[3.1.1]heptane ring system. These derivatives have the amide moiety, in contrast to those with the bicyclo[2.2.1]heptane ring system, which have the sulfonamide group. The derivatives having the 6,6 dimethylbicyclo[3.1.1]heptane ring also exhibited strong activity in PGD(2) receptor binding and cAMP formation assays. In in vivo assays such as allergic rhinitis, conjunctivitis, and asthma models, these series of derivatives showed excellent pharmacological profiles. In particular, compound 45 also effectively suppressed eosinophil infiltration in allergic rhinitis and asthma models. This compound (45, S-5751) is now being developed as a promising alternative antiallergic drug candidate. PMID- 12773049 TI - Structure-activity relationships for 4-nitrobenzyl carbamates of 5 aminobenz[e]indoline minor groove alkylating agents as prodrugs for GDEPT in conjunction with E. coli nitroreductase. AB - Twelve substituted 4-nitrobenzyl carbamate prodrugs of the 5-aminobenz[e]indoline class of DNA minor groove alkylating agents were prepared and tested as prodrugs for gene-directed enzyme prodrug therapy (GDEPT) using a two-electron nitroreductase (NTR) from E. coli B. The prodrugs and effectors were tested in a cell line panel comprising parental and transfected human (SKOV/Skov-NTR(neo), WiDr/WiDr-NTR(neo)), Chinese hamster (V79(puro)/V79-NTR(puro)), and murine (EMT6/EMT6-NTR(puro)) cell line pairs. In the human cell line pairs, several analogues bearing neutral methoxyethoxy-, 2-hydroxyethoxy-, or 3-hydroxypropoxy substituted side chains were good substrates for NTR as measured by cytotoxicity ratios, with NTR-ve/NTR+ve ratios similar to the established NTR substrates CB1954 (an aziridinyl dinitrobenzamide) and the analogous bromomustard. Selectivity for NTR decreased with increasing side-chain size or the presence of a basic amine group. Low to modest selectivity was observed in the Chinese hamster-derived cell line pair; however, in the murine EMT6/EMT6-NTR(puro) cell line pair, the above hydroxyalkoxy analogues again showed significant selectivity for NTR. The activity of the 2-hydroxyethoxy analogue was evaluated against NTR expressing EMT6 tumors comprising ca. 10% NTR+ve cells at the time of tumor treatment. A small decrease in NTR+ve cells was observed after treatment, with a lesser effect against NTR-ve target cells, but these effects were not statistically significant and were much less than for the dinitrobenzamides. These results suggest that useful GDEPT prodrugs based on the 4-nitrobenzyl carbamate and 5-aminobenz[e]indoline motifs may be developed if optimization of pharmacokinetics can be addressed. PMID- 12773050 TI - Discovery and structure-activity relationships of novel piperidine inhibitors of farnesyltransferase. AB - A novel piperidine series of farnesyltransferase (FTase) inhibitors is described. Systematic medicinal chemistry studies starting with the lead compound, discovered from a 5-nitropiperidin-2-one combinatorial library, resulted in a potent series of novel FTase inhibitors. We found that all of four substituents of the piperidine core played an important role for FTase inhibition. A 10-fold increase in potency was observed by changing the piperidine-2-one core to the corresponding piperidine core. This class of compounds was found to inhibit farnesyltransferase in a Ras competitive manner. Optical resolution of several potent inhibitors revealed that the (+)-enantiomers showed potent farnesyltransferase inhibition. (+)-8 inhibited FTase with an IC(50) of 1.9 nM. PMID- 12773051 TI - Discovery of substituted N-phenyl nicotinamides as potent inducers of apoptosis using a cell- and caspase-based high throughput screening assay. AB - By applying a novel cell- and caspase-based HTS assay, a series of N-phenyl nicotinamides has been identified as a new class of potent inducers of apoptosis. Through SAR studies, a 20-fold increase in potency was achieved from a screening hit N-(4-methoxy-2-nitrophenyl)pyridine-3-carboxamide (1) to lead compound 6 methyl-N-(4-ethoxy-2-nitrophenyl)pyridine-3-carboxamide (10), with an EC(50) of 0.082 microM in the caspase activation assay in T47D breast cancer cells. The N phenyl nicotinamides also were found to be active in the growth inhibition assay where compound 10 had a GI(50) value of 0.21 microM in T47D cells. More importantly, compound 10 was found to be equipotent in MES-SA cells and paclitaxel-resistant, p-glycoprotein overexpressed MES-SA/DX5 cells. Compounds 1 and 6-chloro-N-(4-ethoxy-2-nitrophenyl)pyridine-3-carboxamide (8), a more potent analogue, were found to arrest T47D cells in G(2)/M phase of the cell cycle followed by induction of apoptosis as measured by flow cytometry. Compound 8, which was more potent than 1 in the caspase activation assay, also was found to be more potent in G(2)/M arrest and apoptosis assay. These data confirm that the cell-based caspase activation assay is useful for screening for inducers of apoptosis, as well as for SAR studies and lead optimization. Upon further characterization, N-phenyl nicotinamides were found to be inhibitors of microtubule polymerization in vitro. The identification of N-phenyl nicotinamides as a novel series of inducers of apoptosis demonstrates that our cell- and caspase-based HTS assay is useful for the discovery and optimization of potentially novel anticancer agents. PMID- 12773052 TI - Novel indolyl aryl sulfones active against HIV-1 carrying NNRTI resistance mutations: synthesis and SAR studies. AB - The potent anti-HIV-1 activities of L-737,126 (2) and PAS sulfones prompted us to design and test against HIV-1 in acutely infected MT-4 cells a number of novel 1- and 3-benzenesulfonylindoles. Indoles belonging to the 1-benzenesulfonyl series were found poorly or totally inactive. On the contrary, some of the 3 benzenesulfonyl derivatives turned out to be as potent as 2, being endowed with potencies in the low nanomolar concentration range. In particular, (2 methylphenyl)sulfonyl (72) and (3-methylphenyl)sulfonyl (73) derivatives showed EC(50) values of 1 nM. Introduction of two methyl groups at positions 3 and 5 of the phenyl ring of 2 furnished derivatives (80 and 83) which showed very potent and selective anti-HIV-1 activity not only against the wt strain, but also against mutants carrying NNRTI-resistant mutations at positions 103 and 181 of the reverse transcriptase gene. PMID- 12773053 TI - Design, synthesis, and evaluation of analogues of 3,3,3-trifluoro-2-hydroxy-2 phenyl-propionamide as orally available general anesthetics. AB - We have recently discovered a novel class of compounds that have oral general anesthetic activity, potent anticonvulsant activity, and minimal hemodynamic effects. The 3,3,3-trifluoro-2-hydroxy-2-phenyl-propionamide (1) demonstrated potent ability to reduce the minimum alveolar concentration (MAC) of isoflurane, with no effects on heart rate or blood pressure at therapeutic concentrations. Analogue 1 also had potent oral anticonvulsant activity against maximal electroshock (MES) and subcutaneous metrazol (scMET) models with a therapeutic index of 10 for MES activity. In this study, we further synthesized nine new racemic analogues and evaluated these compounds for effects on isoflurane MAC reduction and blood pressure. Preliminary data demonstrate potent reduction in the isoflurane MAC for two new compounds. Current mechanistic studies were unrevealing for effects on voltage-gated ion channels as a putative mechanism. Liposomal partitioning studies using (19)F NMR reveal that the aromatic region partitions into the core of the lipid. This partitioning correlated with general anesthetic activity of this class of compounds. Further, compound 1 was used at a concentration of 1 mM and slightly enhanced GABA(A) current in hippocampal neurons at 10 microM. Altogether, 3,3,3-trifluoro-2-hydroxy-2-phenyl-propionamide exhibited excellent oral general anesthetic activity and appears devoid of significant side effects (i.e., alterations in blood pressure or heart rate). PMID- 12773054 TI - Neuromuscular blocking activity and therapeutic potential of mixed tetrahydroisoquinolinium halofumarates and halosuccinates in rhesus monkeys. AB - Structure-activity relationships in rhesus monkeys for a novel mixed-onium class of ultra-short-acting nondepolarizing tetrahydroisoquinolinium neuromuscular blockers (NMBs) are described. Bis-onium chlorofumarate 20a with (1R,2S) benzyltetrahydroisoquinolinium groups was a potent lead compound (ED(95) = 0.079 mg/kg) with an ultra-short duration of NMB effect (7.1 min) and a selectivity index (SI: defined as a ratio of the cardiovascular threshold dose to the ED(95)) similar to that of mivacurium (3). The mean threshold dose for cardiovascular effects with 20a was ca. 20 times its ED(95) value (SI = 20). A novel mixed-onium analogue of 20a was prepared by replacing the benzyltetrahydroisoquinolinium group distal to the fumarate chlorine atom with a (1S,2R) phenyltetrahydroisoquinolinium moiety. The resulting mixed-onium chlorofumarate 24a displayed good NMB potency (ED(95) = 0.063 mg/kg), ultra-short duration of action (5.6 min) and an improved selectivity index (SI = 57). Several other mixed onium derivatives containing octanedioate (25a; ED(95) = 0.103 mg/kg), difluorosuccinate (27c; ED(95) = 0.056 mg/kg), and fluorofumarate (28a; ED(95) = 0.137 mg/kg) linkers were also potent, ultra-short-acting NMBs with good to excellent selectivity index values (SI = 37-96). Octanedioate 25a was longer acting at higher doses compared to difluorosuccinate 27c and chlorofumarate 24a. Durations of NMB effect following a 0.4 mg/kg bolus dose (100% block) of 25a, 27c, and 24a were 16.9, 13.0, and 10.0 min, respectively. Recovery time for mixed onium chlorofumarate 24a following a 1 h continuous infusion at 10-20 microg/kg/min (95-100% block) was ca. 5 min which is similar to that observed following a 0.2 mg/kg bolus dose of this compound and indicates a lack of cummulative effects. Preliminary studies with chlorofumarate 24a in whole human blood revealed that mixed-onium thiazolidine 29 was the major metabolite and that plasma cholinesterases do not play the primary role in duration of NMB effect. The NMB properties of 24a in rhesus monkeys led to its clinical evaluation as a possible alternative to succinylcholine. PMID- 12773055 TI - A short synthesis and biological evaluation of potent and nontoxic antimalarial bridged bicyclic beta-sulfonyl-endoperoxides. AB - The syntheses and in vitro antimalarial screening of 50 bridged, bicyclic endoperoxides of types 9-13 are reported. In contrast to antimalarial trioxanes of the artemisinin family, but like yingzhaosu A and arteflene, the peroxide function of compounds 9-13 is contained in a 2,3-dioxabicyclo[3.3.1]nonane system 6. Peroxides 9 and 10 (R(1) = OH) are readily available through a multicomponent, sequential, free-radical reaction involving thiol-monoterpenes co-oxygenation (a TOCO reaction). beta-Sulfenyl peroxides 9 and 10 (R(1) = OH) are converted into beta-sulfinyl and beta-sulfonyl peroxides of types 11-13 by controlled S oxidation and manipulation of the tert-hydroxyl group through acylation, alkylation, or dehydration followed by selective hydrogenation. Ten enantiopure beta-sulfonyl peroxides of types 12 and 13 exhibit in vitro antimalarial activity comparable to that of artemisinin (IC(50) = 6-24 nM against Plasmodium falciparum NF54). In vivo testing of a few selected peroxides against Plasmodium berghei N indicates that the antimalarial efficacies of beta-sulfonyl peroxides 39a, 46a, 46b, and 50a are comparable to those of some of the best antimalarial drugs and are higher than artemisinin against chloroquine-resistant Plasmodium yoelii ssp. NS. In view of the nontoxicity of beta-sulfonyl peroxides 39a, 46a, and 46b in mice, at high dosing, these compounds are regarded as promising antimalarial drug candidates. PMID- 12773056 TI - Synthesis and antitumor activity of novel O-carbamoylmethyloxime derivatives of radicicol. AB - Radicicol (1), a macrocyclic antifungal antibiotic, is the lead compound of a novel class of heat shock protein 90 (Hsp90) inhibitors that result in the inhibition or degradation of Hsp90-associated proteins, such as v-src and Raf-1 kinases. New O-carbamoylmethyloxime derivatives of 1 were synthesized and evaluated for their in vitro antiproliferative activities against v-src- and K ras-transformed cells and for their inhibitory activity against v-src tyrosine kinase. O-(Piperidinocarbonyl)methyloxime 9b, one of the most potent of these derivatives, exhibited more potent antiproliferative activity than 1 and its hydroxime KF25706 (2) and had an IC(50) of 25 nM for the inhibition of v-src kinase activity. Compound 9b was also found to decrease the Raf-1 protein level of KNRK5.2 cells. Furthermore, compound 9b exhibited significant antitumor activity when tested against MX-1 and A431 xenografts in nude mice. PMID- 12773057 TI - Elicitation, a new window into plant chemodiversity and phytochemical drug discovery. AB - Plant extracts collected from the wild are important sources for drug discovery. However, these extracts suffer from a lack of reproducible bioactivity and chemical composition caused by the highly inducible, variable, and transitory nature of plant secondary metabolism. Here, we demonstrate that exposing roots of hydroponically grown plants to chemical elicitors selectively and reproducibly induced the production of bioactive compounds, dramatically increased the hit rate, and more than doubled the number of plant species showing in vitro activity against bacteria, fungi, or cancer. Elicitation performed under controlled conditions dramatically improves reliability and efficiency of plant extracts in drug discovery while preserving wild species and their habitats. PMID- 12773058 TI - Analysis of 6-(2,2-Dichloroacetamido)chrysene interaction with the hypoxanthine phosphoribosyltransferase from Trypanosoma cruzi. AB - Selective inhibition is needed for drugs targeting the hypoxanthine phosphoribosyltransferase of Trypanosoma cruzi, etiologic agent of Chagas' disease. 6-(2,2-Dichloroacetamido)chrysene, was shown herein to be a selective inhibitor of the trypanosomal enzyme. SAR analysis revealed that the 6-amido moiety was essential, but the dichloroaceto moiety was not essential for achieving the low K(i) for this inhibitor. Understanding the molecular basis for these interactions could facilitate the design of selective inhibitors without a chrysene moiety. PMID- 12773059 TI - Linezolid resistance since 2001: SENTRY Antimicrobial Surveillance Program. AB - BACKGROUND: The oxazolidinone class of antimicrobials has demonstrated remarkable activity against gram-positive cocci. Linezolid has proven to be a first-line therapeutic option for vancomycin-resistant strains. Linezolid clinical trial results and subsequent published case reports cite rare resistance emerging in patients receiving prolonged therapy. OBJECTIVE: To report the initial linezolid resistant organisms from cases obtained through the SENTRY Antimicrobial Surveillance Program, after screening >40,000 gram-positive cocci without resistance between 1998 and 2000. METHODS: During 2001-2002, 8 resistant strains (from 8 different patients) located in 6 states from 7 different participating SENTRY institutions in the US were identified among bloodstream, respiratory, skin and soft tissue, and urinary tract infection isolates of Enterococcus faecalis, Enterococcus faecium, Staphylococcus epidermidis, and Streptococcus oralis. Resistance was detected by reference broth microdilution methods and confirmed by identical results using Etest (AB BIODISK, Solna, Sweden) and the standardized disk diffusion method. RESULTS: Minimum inhibitory concentration (MIC) and disk diffusion tests showed elevated MICs (> or =8 microg/mL) and small inhibitory zone diameters (< or =15 mm) for all strains to both linezolid and the investigational oxazolidinone AZD2563. Vancomycin resistance was detected in 2 of the 8 linezolid-resistant strains. All enterococci and the viridans-group streptococcus (S. oralis) strain showed resistance to erythromycin. E. faecium strains were resistant to penicillins, but susceptible to quinupristin/dalfopristin. Only 3 of the patients had previously received the drug. CONCLUSIONS: Linezolid resistance remains rare, with only 8 isolates among 9833 (0.08%) monitored isolates identified between January 1, 2001, and June 30, 2002. Resistance, however, was no longer limited to enterococci. Clinical laboratories should test linezolid more widely to detect emerging resistance, especially for patients receiving oxazolidinone therapy. Longitudinal surveillance programs are warranted to detect a trend in the development of resistance, determine the molecular mechanism of resistance, and recommend alternative therapies or epidemiologic interventions. PMID- 12773061 TI - Absorption of enteral recombinant human erythropoietin by neonates. AB - BACKGROUND: Erythropoietin (EPO) is present in amniotic fluid, colostrum, and human milk. One possible function of ingested EPO might be to stimulate erythropoiesis. However, it is unclear whether human neonates absorb recombinant human erythropoietin (rhEPO) after oral administration. OBJECTIVE: To determine whether circulating EPO concentrations increase following enteral administration of rhEPO to neonates. METHODS: The study was designed as a 2-center prospective, blinded, randomized, 2 x 2 crossover study, with each infant receiving 1 dose of rhEPO 1000 units/kg and 1 dose of placebo. Serum EPO concentrations were measured at baseline, 2, and 4 hours following study drug administration. The rhEPO and placebo dosing were separated by a mean of 72 hours. Analysis was stratified by gestational age (< or =35 wk, >35 wk) and feeding type (human milk, infant formula). RESULTS: No significant change in serum EPO concentration was identified at 2 or 4 hours following enteral administration of rhEPO. CONCLUSIONS: Enteral administration of a large dose of rhEPO to neonates <4 months of age did not result in increased circulating EPO concentrations at 2 or 4 hours following dosing, regardless of whether it was administered in human milk or infant formula. PMID- 12773060 TI - Factors affecting patient adherence to highly active antiretroviral therapy. AB - OBJECTIVE: To determine the clinical and demographic variables related to adherence to highly active antiretroviral therapy (HAART) in patients treated in our hospital and identify the characteristics of nonadherent patients. METHODS: Outpatients receiving treatment with HAART (n = 283) were asked about variables related to adherence and to complete the APGAR (family support), State-Trait Anxiety questionnaire (STAI) (emotional situation), and IAS (social support) questionnaires. Patients were classified in 2 groups depending on whether adherence was > or =95% or <95%. Adherence was defined as the percentage of dosage forms prescribed that were obtained by the patient at the hospital pharmacy. A multivariate analysis was created to analyze how each significant variable affected adherence. RESULTS: Our data showed significant nonadherence for patients with the following factors: low level of education, unemployed, emotional situation, and abuse of substances including intravenous drugs. All significant variables were included in a logistic regression model to optimize the results. This model considered 4 variables: age (95% CI 0.89 to 0.99), number of antiretroviral drugs (95% CI 1.05 to 2.11), STAI Anxiety/Trait test (95% CI 2.02 to 6.02), and abuse of drugs (95% CI 1.20 to 3.95). CONCLUSIONS: We recommended special intervention to reinforce adherence for younger patients, patients taking a high number of antiretroviral drugs, those who have a history of intravenous drug use, and those with high anxiety status. PMID- 12773062 TI - Topical corticosteroid prescribing patterns following changes in drug benefit status. AB - OBJECTIVE: To examine changes in prescribing patterns for topical corticosteroid products dispensed to elderly patients in Nova Scotia, Canada, after all but 2 combination topical corticosteroid products were removed from the Nova Scotia Seniors' Pharmacare Program benefit list. METHODS: Administrative prescription claims from the Nova Scotia Seniors' Pharmacare Program were used to identify the number and costs of topical corticosteroid, antifungal, antibiotic, and combination corticosteroid products dispensed. Time-series analysis was used to compare the periods before (April 1, 1999-March 31, 2000) and after (April 1, 2000-March 31, 2001) the delisting. RESULTS: In 1999-2000, 26,031 of 103 400 eligible elderly patients (25%) and in 2000-2001, 22,837 of 95,550 eligible elderly (24%) received a prescription for a defined topical product. Nova Scotia Seniors' Pharmacare Program expenditures for all topical products decreased from 11.88 to 10.60 Canadian dollars per beneficiary per year (11%) after the policy revision. Topical combination products decreased from 18% of all topical products dispensed to 14%, while the percentage of potent corticosteroid products dispensed increased from 24% to 27% over the study period. Pre- and post-policy time-trend analysis showed statistically significant increasing trends in cost per beneficiary for all topical products and potent corticosteroid products. Combination corticosteroid products showed no change in trends for costs per beneficiary before, and a slight increasing trend after, the policy revision. CONCLUSIONS: Prescribing of topical corticosteroid combination products in Nova Scotia decreased following the formulary revision. There was an increase in potent topical corticosteroid prescribing. Future study involving evaluation of patient outcomes would be useful. PMID- 12773063 TI - Comparing methods of establishing the aPTT therapeutic range of heparin. AB - BACKGROUND: The American College of Chest Physicians (ACCP) recommends that the activated partial thromboplastin time (aPTT) therapeutic range for unfractionated heparin be defined as the aPTT corresponding to a heparin concentration of 0.3 0.7 micro/mL by heparin anti-factor Xa assay. This recommendation suggests that a therapeutic range defined in this manner should be superior to traditional empiric therapeutic ranges of 1.5-2.5 times the control. A pilot study was conducted to evaluate the ACCP recommendation for heparin monitoring. OBJECTIVE: To compare heparin dosage adjustments guided by a heparin concentration-derived therapeutic range (HCDTR) with those influenced by traditional empiric therapeutic ranges for the aPTT. METHODS: This study was conducted in 2 phases. In phase 1, the various aPTT therapeutic ranges were established and/or defined. The first empiric therapeutic range (E1) was established by performing an aPTT test on healthy volunteers. This E1 was defined as 1.5-2.5 times the mean normal aPTT. A second empiric therapeutic range (E2) was defined as 1.5-2.5 times the patient's baseline aPTT. The aPTT HCDTR had been defined in a previous study as 48-61 seconds. In phase 2, heparin dosage adjustment decisions guided by each empiric range and the HCDTR for the aPTT were compared with heparin dosage adjustment decisions guided by actual heparin concentrations. Decisions were in agreement when both the aPTT result and plasma heparin concentration indicated the same dosage change. Forty patients had a bedside aPTT determined prior to receiving continuous infusion heparin and again within 48 hours of heparin initiation. Plasma heparin concentration by anti-factor Xa assay was performed on the blood samples obtained after heparin initiation. Heparin dosage adjustment decisions were evaluated by determining the agreement of each aPTT test result with the corresponding plasma heparin concentration. An overall level of agreement (defined as the % of decisions that were in agreement) for each aPTT therapeutic range was determined. RESULTS: The level of agreement in dosage adjustment decisions between heparin concentration and E1, E2, and HCDTR was 28/40 (70%), 28/39 (72%), and 23/40 (58%), respectively (p = 0.34). Heparin dosage adjustment decisions based on an aPTT HCDTR did not significantly differ from heparin dosage adjustment decisions guided by traditional empiric therapeutic ranges for a bedside aPTT. CONCLUSIONS: This pilot study showed similar heparin dosage adjustment decisions using an empiric aPTT therapeutic range versus a heparin concentration-derived aPTT therapeutic range. PMID- 12773065 TI - Stability of cefepime hydrochloride in AutoDose Infusion System bags. AB - OBJECTIVE: To evaluate the physical and chemical stability of cefepime (as the hydrochloride) 1 g/100 mL and 4 g/100 mL admixed in NaCl 0.9% injection and packaged in AutoDose Infusion System bags. DESIGN: Triplicate test samples of cefepime hydrochloride in NaCl 0.9% injection were packaged in ethylene vinyl acetate plastic containers, AutoDose bags, designed for use in the AutoDose Infusion System. Samples were stored protected from light and evaluated at appropriate intervals for up to 7 days at room temperature of approximately 23 degrees C and 30 days under refrigeration at 4 degrees C. Physical stability was assessed using turbidimetric and particulate measurement, as well as visual inspection. Chemical stability was assessed by HPLC. RESULTS: All of the admixtures were initially clear and light yellow when viewed in normal fluorescent room light and with a Tyndall beam. Measured turbidity and particulate content were low initially but increased over time, eventually becoming a yellow or orange precipitate. The higher concentration precipitated earlier; refrigeration slowed precipitation for both test concentrations. HPLC analysis found that the 1-g/100 mL concentration maintained adequate stability for 2 days at 23 degrees C and up to 30 days at 4 degrees C. The 4-g/100 mL concentration maintained adequate stability for 1 day at room temperature and 7 days under refrigeration; however, unacceptable drug loss and precipitation developed after those time points. CONCLUSIONS: Cefepime hydrochloride exhibited physical and chemical stability consistent with previous stability studies. The AutoDose Infusion System bags were not found to affect adversely the physical and chemical stability of this drug. PMID- 12773064 TI - Assessment of an age-adjusted warfarin initiation protocol. AB - OBJECTIVE: To examine the time taken to reach a stable international normalized ratio (INR), as well as the incidence of overanticoagulation of an age-adjusted warfarin initiation protocol. METHODS: Inpatients and outpatients commencing warfarin therapy at 2 teaching hospitals were dosed according to the age-adjusted protocol. Data were collected prospectively. MAIN OUTCOME MEASURES: Time to reach a stable INR of 2-3 and the number of patients experiencing an INR > or =4 during the first week of warfarin therapy. RESULTS: Seventy-three patients were assessed; at the completion of the 4-day titration protocol, 63% had achieved a stable INR. After an additional 2 days of empiric dosage adjustment by the attending physician, 86% of the subjects demonstrated a stable INR. Five patients (7%) experienced an INR > or =4. These patients had a nonsignificant trend toward a lower plasma albumin level compared with other patients (p = 0.057, Student's t test). The INR-driven dose adjustments on days 3 and 4 of this protocol coped with other variables that have been shown to affect maintenance warfarin dosing. These included weight, gender, pharmacologic factors affecting clearance, and the presence of certain predesignated risk factors. CONCLUSIONS: The age-adjusted dosing protocol rapidly achieved a stable INR with minimal overanticoagulation. Patients with low serum albumin levels (<3.0 g/dL) may be sensitive to the effects of warfarin during the loading phase. PMID- 12773066 TI - Rhabdomyolysis causing AV blockade due to possible atorvastatin, esomeprazole, and clarithromycin interaction. AB - OBJECTIVE: To report rhabdomyolysis (RML) causing third-degree atrioventricular block secondary to a possible interaction between atorvastatin, esomeprazole, and clarithromycin. CASE SUMMARY: A 51-year-old white woman presented to the emergency department with severe weakness, near syncope, shortness of breath, and chest pain. On admission, her electrocardiogram demonstrated bradycardia (40 beats/min) and third-degree heart block. A creatine kinase (CK) level was >7000 U/L. Her medication history was significant for long-term use of atorvastatin (>1 y), a 6-week history of esomeprazole use, and three 500-mg doses of clarithromycin just prior to admission. Her symptoms of weakness, shortness of breath, and chest pain coincided with starting the esomeprazole. During her hospitalization, the woman required pacemaker placement and her CK continued to rise to >40,000 U/L. Screening for other causes of RML, such as thyrotoxicosis, infection, and immune or hepatic diseases, was negative. She gradually improved over a 26-day hospitalization. DISCUSSION: This is a case of RML resulting in third-degree atrioventricular blockade. An objective causality assessment of the adverse reaction via the Naranjo probability scale revealed a probable association with atorvastatin and a possible association with esomeprazole and clarithromycin. The pharmacokinetic profiles of these agents suggest that a possible contribution to this reaction was P-glycoprotein (PGP) inhibition by esomeprazole altering atorvastatin's normally significant first-pass clearance. CONCLUSIONS: PGP drug interactions with atorvastatin and other hydroxymethylglutaryl coenzyme A reductase inhibitors (statins) may be associated with unreported risks for RML. Further investigation into PGP impact on HMG-CoA appears warranted. PMID- 12773067 TI - Possible cefotaxime-induced Stevens-Johnson syndrome. AB - OBJECTIVE: To report a case of possible cefotaxime-induced Stevens-Johnson syndrome (SJS). CASE SUMMARY: A 72-year-old woman with an upper urinary tract infection developed erosions and blisters on the skin and the mucous membranes, as well as fever and prostration, soon after the administration of cefotaxime. This presentation is consistent with the features of SJS. Resolution of the clinical manifestations was observed after discontinuation of the drug; all other drugs, infections, or immunologic disorders that could have caused this syndrome were carefully excluded. An objective causality assessment revealed that SJS was possibly associated with the use of cefotaxime. DISCUSSION: Although cephalosporins have been associated with an increased risk for SJS and cefotaxime has been suspected of being associated with SJS in a previous case-control study, this is the first full report for cefotaxime-related SJS in the literature. An immunologically mediated reaction may be the underlying mechanism. CONCLUSIONS: Although cefotaxime administration seems to be the underlying cause of the SJS observed in our patient, establishment of a definite causal relationship requires additional cases and supportive data. PMID- 12773068 TI - Milk transfer and neonatal safety of tacrolimus. AB - OBJECTIVE: To report the first case of tacrolimus measurement in human milk following maternal dosing in a woman who breast-fed while taking the medication. CASE REPORT: A 32-year-old white woman who had taken tacrolimus 0.1 mg/kg/d throughout pregnancy contacted the Motherisk Program at 35 weeks' gestation inquiring about the safety of breast-feeding during maternal tacrolimus therapy. After benefit-risk assessment, the mother decided to breast-feed the baby. METHODS: Manually expressed milk samples were collected over 12 hours following the first tacrolimus dose of the day; pre-dosing and 1-hour post-dosing blood concentrations were also determined. The samples were analyzed for tacrolimus by tandem-mass spectrometry. Breast milk and blood samples were collected at steady state. RESULTS: The highest and mean concentrations of tacrolimus in milk were 0.57 and 0.429 ng/mL, respectively. From these measurements, the exclusively breast-fed infant would ingest, on average, 0.06 micro g/kg/d, which corresponds to 0.06% of the mother's weight-adjusted dose. Given the low oral bioavailability of tacrolimus, the maximum amount the baby would receive is 0.02% of the mother's weight-adjusted dose. The milk-to-blood ratios of tacrolimus at pre-dosing and 1 hour post-dosing concentrations were calculated to be 0.08 and 0.09, respectively. At 2.5 months of age, the infant was developing well both physically and neurologically. COMMENT: This report is the first to measure tacrolimus concentrations in established human milk using tandem-mass spectrometry to detect drug while the infant was exclusively breast-fed by the mother, and in which the infant's growth and development were reported. CONCLUSIONS: Our results suggest that maternal therapy with tacrolimus for liver transplant may be compatible with breast-feeding. PMID- 12773069 TI - Valproic acid-induced neutropenia. AB - OBJECTIVE: To report a case of severe neutropenia caused by valproic acid (VPA). CASE SUMMARY: A 56-year-old white woman with an infectious brain abscess causing tonic-clonic seizure activity was treated with VPA. She developed severe neutropenia after 2 days of VPA therapy. The absolute neutrophil count reached a nadir of 47 cells/mm(3) during VPA use and returned to normal upon its discontinuation. DISCUSSION: VPA is considered to be a well-tolerated antiepileptic drug. While neutropenia has been reported, it has been mild and transient. This patient developed severe neutropenia during effective treatment with VPA, making her significantly susceptible to infection. The Naranjo probability scale indicates VPA as the probable cause of neutropenia in this case. CONCLUSIONS: This report of severe neutropenia caused by VPA emphasizes the importance of monitoring complete blood cell counts during therapy with this agent. PMID- 12773070 TI - Enoxaparin-induced retroperitoneal hematoma. AB - OBJECTIVE: To report 2 cases of retroperitoneal hematoma in elderly patients receiving enoxaparin. CASE SUMMARIES: Two white men, aged 70 and 71 years, received enoxaparin 80 mg subcutaneously twice a day for 8 and 4 days, respectively. Baseline hemoglobin and hematocrit values were 9.5 g/dL and 28.9% and 11.2 g/dL and 32.8%, respectively. In both cases, after the hemoglobin and hematocrit values decreased to 6.6 g/dL and 20.4% and 5.1 g/dL and 15.2%, respectively, a computed tomography scan revealed a retroperitoneal hematoma. DISCUSSION: Enoxaparin is a frequently used anticoagulant. Major bleeding episodes are reported to occur at a rate of up to 5.2%. Factors that increase the risk of bleeding in patients receiving enoxaparin are the use of high doses of enoxaparin, advanced age, renal impairment, and the concomitant use of drugs affecting hemostasis. Both of these patients received relatively high doses of 0.80 and 0.94 mg/kg subcutaneously twice a day, were elderly, and had mild renal impairment; 1 received aspirin concomitantly, while the other received aspirin 4 days prior and warfarin 1 day prior to bleeding. CONCLUSIONS: There are very few published reports implicating enoxaparin as a factor in retroperitoneal hematoma. It is hoped that the addition of these 2 cases to the medical literature creates more awareness that retroperitoneal hematoma should be considered in the differential diagnosis in patients receiving enoxaparin and experiencing unexplained decreases in hemoglobin and hematocrit. PMID- 12773071 TI - Imipramine-associated hyperpigmentation. AB - OBJECTIVE: To inform clinicians of the potential for severe and persistent facial hyperpigmentation with the long-term use of imipramine. CASE SUMMARY: A 65-year old white male veteran with a history of paranoid schizophrenia was referred to the psychiatry service by a dentist who thought that the patient was both cyanotic and psychotic. The history and biopsy results indicated the possibility of imipramine-associated hyperpigmentation, only the second reported case in a male patient. The presentation was complex, with a history of neuroleptic exposure and multiple signs of parkinsonism. A brain single photon-emission computed tomography scan demonstrated frontal lobe hypermetabolism and bilateral caudate hypermetabolism, which normalized 14 months later. Despite discontinuation of imipramine, the patient continued to appear cyanotic, leading to worsening social isolation. He became known as "the man with the purple face." On his rare ventures outside the home, he was embarrassed by sporadic calls to 911 by persons fearing he was ill. DISCUSSION: Although facial hyperpigmentation secondary to the use of phenothiazines has been reported frequently, it is much less common with imipramine, and is very rare in males. Failure to recognize this adverse reaction led to continuing treatment with imipramine and to an apparently irreversible condition. The brain imaging findings have no link with the hyperpigmenting process, but raise questions about neuroleptic-induced metabolic changes in the brain. CONCLUSIONS: Clinicians need to be aware of rare adverse reactions such as hyperpigmentation, and be prepared to take appropriate and early action to prevent such reactions from becoming irreversible. PMID- 12773072 TI - Carbimazole-related gastroschisis. AB - OBJECTIVE: To report a case of gastroschisis in a newborn secondary to carbimazole exposure in utero. CASE SUMMARY: A 25-year-old white woman was treated for Graves disease with carbimazole throughout pregnancy. A boy was born prematurely by vaginal delivery, with a gastroschisis without associated malformative syndrome. Death occurred in the 25th hour of life after surgical repair. DISCUSSION: Carbimazole is completely metabolized to methimazole after absorption. Carbimazole or methimazole intake during pregnancy has been associated with an increased incidence of scalp aplasia. Abdominal wall defects secondary to carbimazole or methimazole exposure in utero seem to be a rare occurrence. However, other cases of abdominal wall defects have been reported in 4 newborns, 2 of them associated with scalp aplasia. An objective causality assessment revealed that the relationship between the gastroschisis and the exposure to carbimazole in utero was possible. CONCLUSIONS: It is important to emphasize the possible risk of abdominal wall defects in newborns to pregnant women taking carbimazole or methimazole. PMID- 12773073 TI - Anaphylactic reaction to a dietary supplement containing willow bark. AB - OBJECTIVE: To report a case of anaphylaxis resulting from the use of a willow bark-containing dietary supplement in a patient with a history of an aspirin allergy. CASE SUMMARY: A 25-year-old white woman presented to the emergency department of a community teaching hospital with anaphylaxis requiring epinephrine, diphenhydramine, methylprednisolone, and volume resuscitation to which she responded favorably. Medication history revealed that she had ingested 2 capsules of Stacker 2 (NVE Pharmaceuticals, Newton, NJ), a dietary supplement promoted for weight loss, prior to experiencing her initial symptoms. Among other active ingredients, this product contains willow bark. Of significance is that this patient also reported a history of allergy to acetylsalicylic acid. No other causes for anaphylaxis were identified. She continued to receive routine supportive care and the remaining hospital course was uncomplicated. DISCUSSION: Dietary supplements, including herbal products, are used by many individuals who consider them to be inherently safe despite limited regulatory oversight by the Food and Drug Administration. While there may be value to specific botanical ingredients, a potential for adverse effects also exists. The popular product consumed by our patient is used for weight loss and contains willow bark, a source of salicylates. Based on the Naranjo probability scale, it is probable that this case of anaphylaxis was due to this dietary supplement. CONCLUSIONS: The use of any willow bark-containing dietary supplement may present a risk of anaphylactic reaction to patients with a history of allergy to salicylates. Clinicians need to recognize the potential for adverse effects from dietary supplements. PMID- 12773074 TI - Interaction between warfarin and the herbal product quilinggao. AB - OBJECTIVE: To describe a patient with loss of anticoagulation control and bleeding after consumption of the combination herbal product quilinggao. CASE SUMMARY: A 61-year-old man who was stable on warfarin therapy regularly consumed a jelly-like herbal product called quilinggao ("essence of tortoise shell"). Five days after the daily consumption of a second brand of quilinggao, he developed easy gum bleeding, epistaxis, and skin bruising with an international normalized ratio (INR) >6.0. Warfarin therapy was temporarily withdrawn until the INR decreased to 1.9. On the day of hospital discharge, he took a third brand of quilinggao against medical advice. Three days later, his INR was 5.2. Warfarin therapy was again temporarily withheld and the patient counseled about an apparent herb-warfarin interaction. He could not remember the name of the third brand of quilinggao. DISCUSSION: Quilinggao is a very popular Chinese herbal product. There are many different brands, and the composition of herbal products varies between manufacturers. Chuanbeimu (Fritillaria cirrhosa) in the first brand and beimu (Fritillaria spp.), chishao (Paeoniae rubra, Chinese peony), jinyinhua (Lonicera japonica), and jishi (Poncirus trifoliata) in the second brand of quilinggao have antiplatelet and/or antithrombotic effects. Loss of anticoagulation control occurred after consumption of the second and third brands of quilinggao, possibly due to the presence of a greater number of interacting herbs. An objective causality assessment revealed that the observed reaction was highly probable to be related to the ingestion of the second and third brands of quilinggao. CONCLUSIONS: Quilinggao contains herbal ingredients that can interact with warfarin. Patients on warfarin therapy should be discouraged from taking herbal medicines, especially preparations that are already known to have antiplatelet and antithrombotic effects. PMID- 12773075 TI - Ezetimibe for management of hypercholesterolemia. AB - OBJECTIVE: To review the primary literature describing the pharmacology of ezetimibe and clinical trials investigating its use in the management of hypercholesterolemia. DATA SOURCES: A MEDLINE search (1966-December 2002) was performed using SCH 48461, SCH 58235, ezetimibe, and 2-azetidinone as key words. English-language articles were identified and the references of these articles were used to further identify pertinent articles and abstracts. Given the paucity of published articles available on ezetimibe, many of the references cited are abstracts. STUDY SELECTION: All acquired articles that discussed the pharmacology, pharmacokinetics, chemistry, and clinical efficacy of ezetimibe were reviewed. DATA EXTRACTION: Articles were selected based on content regarding the medicinal chemistry, pharmacology, and clinical use of ezetimibe. DATA SYNTHESIS: Ezetimibe, approved for use in October 2002, belongs to a new class of antihyperlipidemic agents that uniquely inhibit the absorption of cholesterol by inhibiting the cholesterol transport system located within intestinal cell walls. In humans, ezetimibe reduced cholesterol absorption by >50%. In clinical trials, ezetimibe 10 mg/d reduced low-density lipoprotein cholesterol (LDL-C) by approximately 18% and further enhanced the LDL-C-lowering effect of statin medications by an additional 15-20%. In addition, ezetimibe lowered triglycerides about 5% and increased high-density lipoprotein cholesterol (HDL-C) approximately 3%. Ezetimibe is well tolerated. At present, no serious adverse effects have been directly attributable to ezetimibe. CONCLUSIONS: Based on the data currently available, it appears that ezetimibe has a potential role in the treatment of primary hypercholesterolemia; however further data are needed to determine its long-term tolerability and efficacy. The potential roles for ezetimibe include its concurrent use with a statin to further enhance the lowering of LDL-C. Other possible roles for ezetimibe include its concurrent use with a statin to permit a lowering of statin dosage to avoid statin-related complications or its use as monotherapy to treat hypercholesterolemia when statin use cannot be tolerated or is contraindicated. Outcome data demonstrating that cardiovascular morbidity and/or mortality are reduced by ezetimibe therapy have yet to be generated. PMID- 12773076 TI - Tenofovir disoproxil fumarate. AB - OBJECTIVE: To review the pharmacology, virology, pharmacokinetics, efficacy, safety, resistance profile, and clinical use of tenofovir disoproxil fumarate. DATA SOURCES: A MEDLINE search was performed (1966-August 2002) using the following terms: tenofovir, tenofovir disoproxil fumarate, PMPA (9-(R)-[2 (phosphonomethoxy)propyl]adenine), and Viread. Abstracts from HIV-related meetings were reviewed. DATA EXTRACTION AND STUDY SELECTION: Publications and meeting abstracts regarding tenofovir were reviewed. The most recent and pertinent items were included. DATA SYNTHESIS: Tenofovir disoproxil fumarate is a nucleotide prodrug that is diphosphorylated to its active moiety, tenofovir diphosphate. In this form, tenofovir acts as a reverse transcriptase inhibitor to inhibit HIV-1 replication. In clinical trials, tenofovir was effective at suppressing HIV-1 RNA and boosting CD4+ cell counts. Tenofovir has a long intracellular half-life, which permits once-daily dosing. Since tenofovir does not interact with the cytochrome P450 pathway, it exhibits minimal drug interactions, with the exception of didanosine. Compared with other reverse transcriptase inhibitors, tenofovir may have advantages in terms of toxicity and medication adherence profiles. Ongoing studies are also analyzing tenofovir's activity against hepatitis B virus. CONCLUSIONS: Tenofovir has been shown to be active against HIV-1 in combination with other antiretrovirals. The drug's benefit as a single-agent intensifier of highly active antiretroviral therapy in treatment-experienced patients has been established, and preliminary data for treatment-naive patients are encouraging. PMID- 12773077 TI - Glycoprotein IIb/IIIa receptor inhibitors in percutaneous coronary intervention and acute coronary syndrome. AB - OBJECTIVE: To review the contemporary role of the glycoprotein (GYP) IIb/IIIa receptor inhibitors abciximab, eptifibatide, and tirofiban in patients undergoing percutaneous coronary intervention (PCI) and those with an acute coronary syndrome (ACS), and to provide an algorithm based on currently available evidence for specific agents. DATA SOURCES: Primary articles were identified by a MEDLINE search (1966-January 2003); references cited in these articles provided additional resources. STUDY SELECTION AND DATA EXTRACTION: All of the articles identified from data sources were considered for relevant information; this article primarily addresses large, controlled or comparative studies, and meta analyses. DATA SYNTHESIS: The role of GYP IIb/IIIa inhibitors in patients undergoing PCI and those with ACS has progressed markedly. To date, abciximab has the most robust data in patients undergoing PCI, particularly high-risk individuals. In PCI patients with lower risk (e.g., elective stenting), eptifibatide is a reasonable first-line option. Data do not support tirofiban for routine use in patients undergoing PCI. For individuals with signs and symptoms of ACS, specifically unstable angina or non-ST-segment elevation myocardial infarction (MI), eptifibatide or tirofiban is recommended in high-risk patients when a conservative approach is used (PCI is not planned). Abciximab is not recommended in this situation. In patients with ST-segment elevation MI (STEMI), abciximab is the only GYP IIb/IIIa inhibitor evaluated in large, well-designed investigations. For medical management in combination with a fibrinolytic agent, the role of abciximab remains unclear. For patients undergoing primary PCI for the management of STEMI, the available evidence supports the use of abciximab, albeit further investigation is warranted. CONCLUSIONS: The role of GYP IIb/IIIa inhibitors in clinical cardiology continues to evolve. Choice of the agent depends on situation of use, patient-specific characteristics and risk stratification, and, in the case of ACS, chosen management strategy (medical management or intervention). PMID- 12773078 TI - Chromium as adjunctive treatment for type 2 diabetes. AB - OBJECTIVE: To review the chemistry, pharmacology, efficacy, and safety of trivalent chromium in the treatment of type 2 diabetes and hyperlipidemia. DATA SOURCES: The English literature was searched from 1966 through May 2002 using MEDLINE, International Pharmaceutical Abstracts, and EMBASE. The key words included chromium, glucose, lipids, and diabetes. Pertinent references from review articles and studies were used as additional sources. DATA SYNTHESIS: Trivalent chromium is an essential nutrient and has a key role in lipid and glucose metabolism. Supplementation with chromium does not appear to reduce glucose levels in euglycemia. It may, however, have some efficacy in reducing glucose levels in hyperglycemia. The effects of chromium on lipid levels are variable. Chromium in doses <1000 microg/d appears to be safe for short-term administration. Kidney function and dermatologic changes need to be monitored. CONCLUSIONS: Chromium appears to be a safe supplement and may have a role as adjunctive therapy for treatment of type 2 diabetes. Additional large-scale, long term, randomized, double-blind studies examining the effect of various doses and forms of chromium are needed. PMID- 12773079 TI - Combination ACE inhibitors and angiotensin II receptor blockers for hypertension. AB - OBJECTIVE: To review data concerning combined angiotensin-converting enzyme (ACE) inhibitor and angiotensin II receptor blocker (ARB) therapy for hypertension. DATA SOURCES: MEDLINE (1966-April 2003), IPA (1970-April 2003), and EMBASE (1974 April 2003) with search terms of ACE inhibitor, angiotensin receptor blocker, essential hypertension, and combination therapy. DATA SYNTHESIS: ACE inhibitors provide incomplete blockade of the renin-angiotensin system, sometimes leading to loss of blood pressure control. Addition of ARBs may in theory further reduce blood pressure. Studies of combined ACE inhibitor and ARB therapy for managing hypertension were evaluated. CONCLUSIONS: While studies have shown statistically significant blood pressure reductions with ACE/ARB combination therapy, clinical significance is lacking. Further trials are needed before routine use of the combination can be recommended. PMID- 12773080 TI - Role of fluoxetine in anorexia nervosa. AB - OBJECTIVE: To evaluate the efficacy of fluoxetine in the treatment of anorexia nervosa. DATA SOURCES: Literature obtained through searching MEDLINE (1966 January 2003). DATA SYNTHESIS: Studies conducted on the efficacy of fluoxetine in treating anorexia nervosa have conflicting results. Study design and methodology should be carefully evaluated. CONCLUSIONS: Fluoxetine played a role in the reduction of symptoms of obsessive-compulsive disorder and depression in anorexic patients. While psychotherapy, nutritional therapy, and behavioral therapy should be the mainstays of treatment, fluoxetine should be considered as an option to prevent relapse or to treat associated symptoms of anorexia nervosa following adequate weight restoration as a part of maintenance therapy and not as a primary or acute therapy. Although fluoxetine appears to be promising for the treatment of patients with anorexia nervosa, further studies need to be performed with a larger sample size and/or better design. PMID- 12773081 TI - Quality assessment programs for dietary supplements. AB - BACKGROUND: In the US, consumers and healthcare professionals have been forced to depend on self-regulation by the dietary supplement industry for product quality, consistency, potency, and purity, a system that has yielded mixed results at best. OBJECTIVE: To characterize the current state of product quality verification in the dietary supplement industry and review recent developments and future plans to improve supplement safety and regulation. METHODS: Studies describing the quality of dietary supplements are reviewed, along with recent Food and Drug Administration (FDA) plans to improve supplement regulation. Several independent dietary supplement quality certification programs are described and evaluated. RESULTS: Quality control standards for dietary supplements run the gamut from good to nonexistent. Contamination, false labeling, and incomplete labeling are not uncommon problems--as are significant discrepancies in disintegration, dissolution, and in vitro release characteristics of various dietary supplements. The need for improved regulation has led to plans for increased FDA involvement through enforcement of good manufacturing practices and truths in labeling. Meanwhile, independent product certification companies have emerged and provide some reassurances; however, they possess several limitations as well. CONCLUSIONS: Until proposed FDA initiatives are finalized and implemented, certification programs can help provide assurances of safety and assist in product selection. Although they do not testify to effectiveness (and may even inadvertently mislead some consumers), certification programs may help curb what has been called "the Wild West nature of the dietary supplement industry." PMID- 12773082 TI - Use of OTC and herbal products in patients with cardiovascular disease. AB - BACKGROUND: The use of nonprescription and herbal products by the public is rising, resulting in an increased potential for adverse reactions or drug interactions in cardiac patients. OBJECTIVE: To describe the utilization patterns for nonprescription medications and herbal products in patients with cardiovascular disease across Canada. METHODS: Patients admitted to 8 teaching hospitals during the winter of 1998/1999 were interviewed by a pharmacist using a structured survey instrument. RESULTS: Interviews were conducted with 306 patients (mean age 66 y; 60% men). The majority (74%) had coronary artery disease; however, hypertension, congestive heart failure, and arrhythmias were also common. The most common product categories used were pain relievers (51%), single-entity vitamin/mineral (38%), multivitamin/mineral (23%), antacids (21%), laxatives (17%), and herbals (17%). As compared with western (28%) and central Canada (26%), fewer patients in the Atlantic region (11%) reported daily use of multivitamin/mineral products. Overall, the usage of specific single-entity vitamin/mineral products was most commonly vitamin E (24%), vitamin C (16%), calcium (9%), and B vitamins (8%). Central Canada reported the highest rates (25%) of daily or weekly use of herbal products. The most common herbal products used were garlic (13%), cayenne pepper (2%), and ginseng (2%). More than half of the patients consulted with their pharmacist at least occasionally regarding the use of these products. CONCLUSIONS: Canadian patients with cardiovascular disease commonly report the use of herbal products and vitamins. Allied health professionals need to be aware of the widespread use of these products and their potential for adverse reactions and drug interactions. PMID- 12773083 TI - Initiating warfarin therapy. PMID- 12773084 TI - Resistance to linezolid. Are we surprised? How hard should we look? PMID- 12773085 TI - Hormonal birth control and pregnancy: a comparative analysis of thromboembolic risk. PMID- 12773086 TI - Quetiapine impact on quality of life in patients with schizophrenia. PMID- 12773087 TI - Pediatric fatality due to anticonvulsant hypersensitivity syndrome. PMID- 12773088 TI - Obese man treated with drotrecogin alfa (activated). PMID- 12773089 TI - Nonketotic hyperosmolar syndrome from olanzapine, lithium, and valproic acid cotreatment. PMID- 12773092 TI - Effects of euglycaemic and hypoglycaemic hyperinsulinaemia on sympathetic and parasympathetic regulation of haemodynamics in healthy subjects. AB - The effects of hypoglycaemia during hyperinsulinaemia, occurring under various pathophysiological conditions, on the cardiovascular regulatory system and vasculature are largely unknown. The aim of the present study was to investigate regulatory and haemodynamic responses to acute hyperinsulinaemia and consequent hypoglycaemia in 18 healthy subjects. Blood sampling and 5 min ECG and blood pressure recordings were performed at baseline and during the euglycaemic and hypoglycaemic phases of a hyperinsulinaemic clamp. Heart rate variability (HRV) and blood pressure variability (BPV) were assessed by using power spectral analysis, and baroreflex sensitivity (BRS) was assessed using the cross-spectral method. Stroke volume was assessed from the non-invasive blood pressure signal by the arterial pulse contour method. Euglycaemic hyperinsulinaemia did not change plasma catecholamine concentrations, HRV, BPV, BRS, heart rate, blood pressure, stroke volume, cardiac output or peripheral resistance. However, hyperinsulinaemic hypoglycaemia resulted in an 11.7-fold increase in the plasma adrenaline concentration (from 0.19+/-0.03 to 1.68+/-0.32 nmol/l; P <0.001), and a modest 1.3-fold increase in the plasma noradrenaline concentration (from 1.74+/ 0.22 to 2.02+/-0.19 nmol/l; P <0.05) compared with baseline. Furthermore, we observed significant decreases in diastolic blood pressure (from 68+/-3 to 60+/-3 mmHg; P <0.05) and peripheral resistance (from 24.1+/-1.2 to 18.5+/-1.1 mmHg.min( 1) x l(-1); P <0.01). Stroke volume and cardiac output increased markedly from the euglycaemic to the hypoglycaemic period only ( P <0.01 for both). Hypoglycaemia did not influence HRV, BPV or BRS. Our findings indicate that hyperinsulinaemic hypoglycaemia is characterized by a significant increase in the plasma adrenaline concentration and by decreases in peripheral resistance and blood pressure. Counter-regulation during hyperinsulinaemic hypoglycaemia involves selective adrenomedullary sympathetic activation, and does not influence cardiac parasympathetic regulation or baroreflex control of heart rate. PMID- 12773093 TI - Association between the cholesteryl ester transfer protein TaqI-detectable B polymorphism and low high-density lipoprotein cholesterol concentration in Saudis. AB - Plasma concentrations of HDL (high-density lipoprotein) cholesterol are low in the Saudi Arabian population. A B polymorphism at the CETP (cholesteryl ester protein transfer) locus that is detectable with the restriction enzyme Taq I is a genetic determinant of the plasma HDL cholesterol concentration. We assessed the relationship between the Taq I B CETP polymorphism and lipid and apolipoprotein concentrations in a study sample of 335 Saudi residents. The Taq I B1 and B2 allele frequencies were 0.54 and 0.46 respectively, similar to those in other populations. HDL cholesterol levels in B2B2 homozygotes were significantly higher than in B1B1 homozygotes [1.01 (0.3) compared with 0.92 (0.2) mmol/l; mean (S.D.); P=0.03]. There was also a significant difference between the B2B2 and B1B1 homozygotes with regard to apolipoprotein AI concentration [123.6 (16.4) compared with 113.7 (13.9) mg/dl; P=0.04]. This genetic variation was independent of metabolic risk factors known to influence HDL cholesterol levels. The allele frequency of the Taq I B CETP polymorphism and its relatively modest impact on HDL cholesterol concentrations argue against an important role for this allele, or for strongly linked loci, in determining the low levels of HDL cholesterol seen in the Saudi population. PMID- 12773094 TI - A role for Sec1/Munc18 proteins in platelet exocytosis. AB - A critical aspect of haemostasis is the release of clot-forming components from the three intra-platelet stores: dense-core granules, alpha granules and lysosomes. Exocytosis from these granules is mediated by soluble proteins [N ethylmaleimide-sensitive fusion protein (NSF) and soluble NSF attachment proteins (SNAPs)] and integral membrane proteins [vesicle and target SNAP receptors (v- and t-SNAREs)]. Three Sec1/Munc18 proteins (SM proteins) are present in platelets (Munc18a, Munc18b and Munc18c) and they bind to and potentially regulate specific syntaxin t-SNAREs. In resting platelets, these SM proteins associate with granules and open canalicular system membranes predominantly but not with the plasma membrane. Munc18a binds to syntaxin 2 alone and does not associate with other members of the core SNARE complex. Munc18b associates with a larger complex that contains synaptosome-associated protein of 23 kDa (SNAP-23) and cellubrevin/vesicle-associated membrane protein 3. Munc18c associates with both syntaxins 2 and 4, with synaptosome-associated protein of 23 kDa (SNAP-23) and with a v-SNARE. On stimulation, most of the platelet SM proteins are still found in membrane fractions. Phosphorylation of each Munc18 increases in thrombin treated cells and phosphorylated Munc18c remains associated with syntaxins 2 and 4, but its affinity for the SNAREs appears to be reduced. To determine the functional role of the platelet SM proteins, we examined the effects of Munc18 based peptides (Munc18a peptide 3 and Munc18c peptide 3). Addition of the peptides to permeabilized platelets inhibits secretion from all three platelet granules. These peptides also inhibit agonist-induced aggregation in saponin permeabilized platelets. These studies demonstrate a clear role for SM proteins in platelet exocytosis and aggregation and suggest a dominant role for Munc18c in all three granule-release events. PMID- 12773096 TI - L-tyrosine prevents aggregation of therapeutic proteins by gamma-irradiation. AB - The present paper describes a protective role of L-tyrosine against aggregation of caeruloplasmin and haemoglobin therapeutic proteins during their sterilization by gamma-irradiation in the aqueous phase. Irradiation of proteins, known to induce their degradation in the presence of oxygen, generates aggregation under oxygen-free conditions. It was found that L-tyrosine present during irradiation in deoxygenated media prevents protein aggregation even at high doses (10 kGy), as asserted by SDS/PAGE and high-performance size-exclusion chromatography. The protective role of L-tyrosine, allowing the gamma-irradiation treatment of therapeutic proteins in solution without conformational alteration, is probably exerted by scavenging oxygen radicals produced by irradiation-induced water radiolysis. It was also found that haemoglobin had a greater stability than caeruloplasmin under gamma-irradiation treatment. PMID- 12773097 TI - CO2 hydration by immobilized carbonic anhydrase. AB - The enzyme carbonic anhydrase (isoform II) from bovine and human erythrocytes was immobilized using different covalent coupling methods on inert matrices. Immobilized carbonic anhydrase may enable concentration of CO2 for Rubisco (ribulose-1,5-bisphosphate carboxylase/oxygenase)-catalysed fixation in bioreactors. In the present study the activity of carbonic anhydrase with respect to hydration of CO2 using soluble and immobilized enzymes was determined. The stability of the immobilization matrix, the properties of the immobilized enzymes subjected to a variation in operation variables and the activity profile with respect to storage are reported. Immobilization imparted greater thermal and storage stability and enhanced reusability. PMID- 12773098 TI - Caeruloplasmin sterilized by gamma-irradiation in the presence of L-tyrosine maintains structural and catalytic characteristics. AB - It was previously shown that protein aggregation induced by gamma-irradiation can be prevented by L-tyrosine, even when irradiation was done in solution at doses as high as 10 kGy. It is now reported that caeruloplasmin irradiated at low to moderate doses appears more resistant to trypsin-mediated proteolysis than native caeruloplasmin. Ceruloplasmin presents an irradiation-dose-dependent decrease of oxidase activity and a slight increase of ferroxidase activity at low irradiation doses (2 kGy), followed by a decrease at high doses (4-8 kGy). In all cases, the catalytic activities are higher when caeruloplasmin is irradiated under tyrosine protection. Irradiated caeruloplasmin in the presence of tyrosine preserves its oxidase and ferroxidase activities. Caeruloplasmin irradiated in solution at a dose of 2-3 kGy and higher, in the presence and absence of L-tyrosine, remained sterile during at least 8 weeks of storage at 4 degrees C, as evaluated by the plate-count-agar method for assessing total viable lactic acid bacteria content after incubation for 48 h at 30 degrees C. PMID- 12773099 TI - Proliferation and functional maturation of Sertoli cells, and their relevance to disorders of testis function in adulthood. AB - Disorders of testicular function may have their origins in fetal or early life as a result of abnormal development or proliferation of Sertoli cells. Failure of Sertoli cells to mature, with consequent inability to express functions capable of supporting spermatogenesis, is a prime example. In a similar way, failure of Sertoli cells to proliferate normally at the appropriate period in life will result in reduced production of spermatozoa in adulthood. This review focuses on the control of proliferation of Sertoli cells and functional maturation, and is motivated by concerns about 'testicular dysgenesis syndrome' in humans, a collection of common disorders (testicular germ-cell cancer, cryptorchidism, hypospadias and low sperm counts) which are hypothesized to have a common origin in fetal life and to reflect abnormal function of Sertoli (and Leydig) cells. The timing of proliferation of Sertoli cells in different species is reviewed, and the factors that govern the conversion of an immature, proliferating Sertoli cell to a mature, non-proliferating cell are discussed. Protein markers of maturity and immaturity of Sertoli cells in various species are reviewed and their usefulness in studies of human testicular pathology are discussed. These markers include anti-Mullerian hormone, aromatase, cytokeratin-18, GATA-1, laminin alpha5, M2A antigen, p27(kip1), sulphated glycoprotein 2, androgen receptor and Wilms' tumour gene. A scheme is presented for characterization of Sertoli-cell only tubules in the adult testis according to whether or not there is inherent failure of maturation of Sertoli cells or in which the Sertoli cells have matured but there is absence, or acquired loss, of germ cells. Functional 'de differentiation' of Sertoli cells is considered. It is concluded that there is considerable evidence to indicate that disorders of maturation of Sertoli cells may be a common underlying cause of human male reproductive disorders that manifest at various life stages. This recognition emphasizes the important role that animal models must play to enable identification of the mechanisms via which failure of proliferation and maturation of Sertoli cells can arise, as this failure probably occurs in fetal life. PMID- 12773100 TI - Maternal and fetal amino acid concentrations and fetal outcomes during pre eclampsia. AB - The pathophysiology of pre-eclampsia is contested, but one hypothesis indicates that it is a heterogeneous condition in which only a subset of affected women bear small-for-gestational age (SGA) babies. In intrauterine growth-restricted (IUGR) infants, placental transport of amino acids is diminished and the resulting decrease in cord-blood amino acid concentrations is thought to contribute to their stunted growth. In contrast, the metabolic syndrome (dyslipidaemia, hyperinsulinaemia, hyperglycaemia, hypertension and obesity) which is associated with high amino acid concentrations is more prevalent in women with pre-eclampsia. The focus of this study was to compare maternal and fetal serum amino acid concentrations during normal pregnancy and pre-eclampsia and to evaluate the associations between the amino acid concentrations and fetal growth. The results indicate that maternal and cord-blood amino acid concentrations were significantly higher in women with pre-eclampsia compared with normal pregnant women and the concentrations were inversely associated with measures of infant growth. Maternal and cord-blood amino acid concentrations were also significantly higher in pre-eclamptic mothers with SGA infants compared with pre-eclamptic mothers whose babies were not SGA. These data indicate that, in contrast to IUGR, pre-eclampsia is associated with enhanced placental amino acid transport or reduced fetal amino acid utilization. Furthermore, the data are consistent with the hypothesis that pre-eclampsia is a heterogeneous disease associated with the metabolic syndrome. PMID- 12773102 TI - Effect of GnRH conjugated to pokeweed antiviral protein on reproductive function in adult male dogs. AB - This study evaluated the effect of a GnRH analogue conjugated to the cytotoxin, pokeweed antiviral protein (PAP), on reproductive function in adult, male dogs. Four dogs received 0.0042 mg GnRH-PAP kg(-1) hourly for 36 h, and four other dogs received 0.1 mg GnRH-PAP kg(-1) as one bolus injection daily for three consecutive days. One dog received a single bolus (0.1 mg x kg(-1)). Three adult male dogs received GnRH without the PAP conjugate, as controls. Twenty-five weeks after the initial treatment, all treated dogs received 0.1 mg GnRH-PAP kg(-1) as a single administration, whereas dogs in the control group received 0.0045 mg kg( 1) of the GnRH analogue. Serum concentrations of testosterone and LH were determined by radioimmunoassay, and testis size was measured for 9 months after treatment. Stimulation tests (5 microg GnRH kg(-1)) were used to evaluate LH release (-15, 0, 30, 60, 90, 120 min), which was assessed by measuring area under the curve. Serum testosterone concentrations were significantly lower (P<0.05) after treatment in the bolus and hourly groups than in the control group. Testosterone concentrations fell to less than 50 pg x ml(-1) in three of four dogs in the bolus group and one of four dogs in the hourly group by week 8-9 after treatment. Basal LH was lower (P<0.05) in the bolus and hourly groups than in the control group between weeks 0 and 33 after treatment. Treatment with GnRH PAP reduced (P<0.05) LH release after GnRH stimulation in the bolus and hourly groups compared with the control group. Testis volume was lower (P<0.05) in all treated versus control dogs. In conclusion, administration of the conjugate GnRH PAP at a 25 week interval resulted in a major disruption of reproductive parameters in male dogs; this effect was maintained for 11-12 weeks after a second injection of GnRH-PAP. PMID- 12773095 TI - Principles of interleukin (IL)-6-type cytokine signalling and its regulation. AB - The IL (interleukin)-6-type cytokines IL-6, IL-11, LIF (leukaemia inhibitory factor), OSM (oncostatin M), ciliary neurotrophic factor, cardiotrophin-1 and cardiotrophin-like cytokine are an important family of mediators involved in the regulation of the acute-phase response to injury and infection. Besides their functions in inflammation and the immune response, these cytokines play also a crucial role in haematopoiesis, liver and neuronal regeneration, embryonal development and fertility. Dysregulation of IL-6-type cytokine signalling contributes to the onset and maintenance of several diseases, such as rheumatoid arthritis, inflammatory bowel disease, osteoporosis, multiple sclerosis and various types of cancer (e.g. multiple myeloma and prostate cancer). IL-6-type cytokines exert their action via the signal transducers gp (glycoprotein) 130, LIF receptor and OSM receptor leading to the activation of the JAK/STAT (Janus kinase/signal transducer and activator of transcription) and MAPK (mitogen activated protein kinase) cascades. This review focuses on recent progress in the understanding of the molecular mechanisms of IL-6-type cytokine signal transduction. Emphasis is put on the termination and modulation of the JAK/STAT signalling pathway mediated by tyrosine phosphatases, the SOCS (suppressor of cytokine signalling) feedback inhibitors and PIAS (protein inhibitor of activated STAT) proteins. Also the cross-talk between the JAK/STAT pathway with other signalling cascades is discussed. PMID- 12773101 TI - E- and N-cadherin expression and distribution during luteinization in the rat ovary. AB - Cadherins, a family of Ca(2+)-dependent cell adhesion molecules, play an important role in ovarian tissue remodelling processes. The aim of this study was to examine the expression pattern of E- and N-cadherin in rat preovulatory follicles, luteinizing follicles and corpora lutea. Immature female rats were treated with equine chorionic gonadotrophin (eCG) to promote preovulatory follicle development. At 48 h after eCG treatment, the rats were injected with an ovulatory dose of hCG. Ovaries were analysed by western blot analysis and immunofluorescence for E- and N-cadherin expression at 48 h after eCG injection, and at 24 and 72 h after hCG injection. Ovaries of cyclic adult rats were examined to assess whether the changes in the expression pattern of cadherin were in agreement with those of the gonadotrophin-treated rats. Finally, expression of E-cadherin in luteinizing granulosa cells in vitro was assessed by RT-PCR and western blot analysis. Immunofluorescence results indicate that E-cadherin is expressed in the theca-interstial cells surrounding preovulatory follicles. N cadherin expression is prominent in the membrana granulosa of these follicles. The initiation of luteinization with hCG leads to a decreased expression of N cadherin in the membrana granulosa, whereas expression of E-cadherin starts within the luteinizing follicle. Both cadherins are prominently expressed in the fully formed corpus luteum at 72 h after hCG treatment. Immunofluorescence results revealed that the patterns of E- and N-cadherin expression in the gonadotrophin-treated rats were similar to those of the cyclic adult rats. Western blot analysis reflected similar changes for N-cadherin in the ovaries of both the cyclic adults and gonadotrophin-treated rats; however, they were different in E-cadherin expression. The expression of E-cadherin mRNA and protein was induced in vitro in luteinized granulosa cells. These results support the hypothesis that modulation of cadherin expression is an integral component of remodelling processes, including corpus luteum formation, in the ovary. The results also indicate that expression of E- and N-cadherin in granulosa-lutein cells appear to be under hormonal control. PMID- 12773103 TI - Regulation of nitric oxide synthase isoforms and role of nitric oxide during prostaglandin F2alpha-induced luteolysis in rabbits. AB - Total activity of nitric oxide synthase (NOS) and the gene expression of both endothelial NOS (eNOS) and inducible NOS (iNOS) isoforms in corpora lutea of pseudopregnant rabbits were examined during prostaglandin F(2alpha) (PGF(2alpha)) induced luteolysis. Corpora lutea were collected at 0, 6, 12, 24 and 48 h after an injection of PGF(2alpha) at day 9 of pseudopregnancy. At 12 h after PGF(2alpha) administration, luteal mRNA encoding eNOS decreased (Por=5.7 mm (t = 0 h). Oestradiol antiserum (100 ml) was given in a single injection at t = 12 h. Follicles of the wave were defined as F1 (largest) and F2, according to the diameter at each examination. Blood samples were collected at 12 h intervals during t = 0-72 h. Treatment with progesterone lowered the circulatory concentrations of LH by 12 h after the start of treatment (P < 0.05), and concentrations remained low compared with those of controls during the treatment period. Treatment with oestradiol antiserum had no effect on LH. Both progesterone and the antiserum treatments increased the FSH concentrations compared with controls (P < 0.05), which supports the first hypothesis. The interval from t = 0 h to the beginning of deviation was longer in the progesterone- (51.0 +/- 7.6 h; P < 0.06) and antiserum (51.4 +/- 6.3 h; P < 0.05) treated groups than in the controls (38.0 +/- 3.7 h), which supports the second hypothesis. There was no difference among groups in the diameters of F1 and F2 at deviation. Reduced diameter (P < 0.05 or P < 0.06) of both F1 and F2 occurred in both the progesterone- and antiserum-treated groups at t = 36 h and 48 h, compared with controls. Follicle retardation occurred in both the progesterone- and antiserum-treated groups despite the high FSH concentrations, whereas LH was altered only in the progesterone-treated group. Therefore, the follicle effect can be attributed to inadequate intrafollicular oestradiol. This interpretation implies a functional local role for oestradiol in the deviation process, independent of the systemic negative effect on FSH. PMID- 12773108 TI - Production of capsular material by equine trophoblast transplanted into immunodeficient mice. AB - A novel xenogeneic transplantation approach was used to determine whether it is embryonic or maternal tissue that produces the material that gives rise to the mucin-like glycoprotein of the equine embryonic capsule. Endometrial biopsy samples and conceptuses from six mares at days 13-15 after ovulation were prepared as 1 mm(3) grafts of endometrium, trophoblast and capsule for transplantation, alone or in combination, into various sites in 88 immunodeficient (severe combined immunodeficient or RAG2/gamma(c) double mutant) mice. The overall recovery rate of grafts was over 50%, reaching 100% with experience and use of the renal subcapsular space exclusively. Periodic acid Schiff (PAS) staining demonstrated capsule-like extracellular glycoprotein secretions at the graft site in 11 of 22 sites examined. Strong PAS-positive reactions (5-7 microm thick) were found in four of six sites containing trophoblast alone, five of six endometrium plus trophoblast sites, and zero of eight grafts of endometrium alone. Two recovered grafts of capsule were also PAS positive. The secreted glycoprotein was identified as equine embryonic capsule material by using a monoclonal antibody (mAb) specific to equine capsule (mAb OC 1) in two experiments. In the first, in cryosections, this antibody bound to 19 of 19 recovered trophoblast graft secretions (including those in 12 from mice that had not received endometrium at any site), ten of ten recovered endometrium plus trophoblast grafts, and zero of 12 recovered endometrial grafts from mice in which trophoblast had been grafted to the same site or another site in the same mouse. In the second experiment, in paraformaldehyde-fixed sections of grafts from 11 mice, specific staining, identical to that shown by grafted capsule, was obtained with grafts of trophoblast (both alone and in combination with endometrium) but not with grafts of endometrium. These results support the contention that trophoblast is the principal source of equine embryonic capsule. In addition, they demonstrate that xenogeneic grafting is a useful means of culturing endometrium and conceptus tissues outside the mare when in vitro techniques do not suffice. PMID- 12773109 TI - Changes in oocyte plasma membrane binding sites on boar spermatozoa with capacitation and acrosome reactions. AB - The objective of this study was to determine the localization and distribution of oocyte plasma membrane binding sites on capacitated and acrosome-reacting live boar spermatozoa. Localization of oocyte plasma membrane binding sites on boar spermatozoa was determined with fluorescence microscopy and population distribution was examined with flow cytometry. The number of spermatozoa with oocyte plasma membrane bound to the equatorial segment and postacrosomal region of the sperm head significantly increased with capacitation. Equatorial segment labelling further increased with induced acrosome reactions. When the population distribution of oocyte plasma membrane binding sites on live boar spermatozoa was analysed, the percentage of spermatozoa with bound oocyte plasma membrane significantly increased after capacitation compared with that of washed spermatozoa. Binding of oocyte plasma membrane did not increase in control spermatozoa incubated under non-capacitating conditions and was not correlated with the percentage of dead spermatozoa. A change in localization of oocyte plasma membrane binding sites on the sperm head was demonstrated using fluorescence microscopy and an increase in oocyte plasma membrane binding sites after capacitation was shown using flow cytometry. PMID- 12773110 TI - Association of segmentation of the epididymal interstitium with segmented tubule function in rats and mice. AB - The epithelium of the epididymal tubule has different biological functions in different regions of the tubule. Each region is further organized into lobules or intra-regional segments surrounded by connective tissue septa (CTS). Epididymal segmentation has received little direct attention, yet there is considerable evidence that expression of mRNA and protein often begins or ends precisely at the CTS border of a segment. How such 'on-off' regulation occurs coincident with the passing of the tubule from one segment to the next is unknown. This study examined the segmentation of epididymides in rats and mice. The average adult Sprague-Dawley rat and C57BL/6 mouse caput, corpus and cauda epididymides has seven, two and four, and three, one and two segments, respectively. The apoptosis response of the caput epididymal epithelium to deprivation of lumicrine factors 24 h after efferent duct ligation in rats and the epididymal expression of a marker protein, beta-galactosidase, in mice were segmented precisely. This validated both at a general response and at a specific protein level that many epididymal functions are regulated within segments. Blue dextran (molecular weight 20000) and erythrocine red (molecular weight 880) dyes infused into the interstitial space of specific segments by micropuncture were retained by the CTS of the segments. In similar micropuncture experiments, [(3)H]H(2)O (molecular weight 18) was able to diffuse into an adjacent segment relatively freely whereas [(14)C]polyethylene glycol (molecular weight 4000) could not. These studies indicate that the interstitium of intra-regional segments is organized into different physiological compartments and that these compartments play a role in regulating the epididymal epithelium. PMID- 12773111 TI - Arrested apoptosis without nuclear fragmentation produced by efferent duct ligation in round spermatids and multinucleated giant cells of rat testis. AB - The apoptotic process evoked by efferent duct ligation in the testes of adult rats was followed for 10 days by differential staining for haematoxylin-eosin, periodic acid-Schiff and a modified trichrome technique in optical microscopy and by ultrastructural localization of acid phosphatase. Round spermatids showed the first effects of efferent duct ligation. At day 3 after ligation, annular clumps of chromatin with typical apoptotic characteristics appeared against the nuclear membrane of these cells. Afterwards, membranous structures and a wide separation between the two layers of the nuclear membrane were observed but nuclear fragmentation did not occur and apoptotic granules were not seen. Cytoplasmic components were also altered, and severely damaged organoids and empty vacuoles lacking acid phosphatase reaction were frequently seen. On day 2 after efferent duct ligation, multinucleated giant cells appeared, which displayed similar characteristics as spermatids and showed no acid phosphatase reaction. Although abnormal spermatids and multinucleated giant cells were surrounded by the cytoplasm of Sertoli cells, neither lysosomal acid phosphatase nor phagocytic activity was detected. It is concluded that efferent duct ligation specifically affects round immature spermatids eliciting a partial nuclear apoptotic response that is not accompanied by autophagic or heterophagic activity and without lysosomal participation in Sertoli cells. PMID- 12773113 TI - Steroidogenic factor-1: a key regulator of gonadotroph gene expression. PMID- 12773114 TI - Tumour necrosis factor alpha: a key regulator of adipose tissue mass. AB - In addition to its established role in the immune system, tumour necrosis factor alpha (TNFalpha) exerts complex regulatory actions on adipose tissue. TNFalpha is produced in and secreted by the adipocyte and thus is in a position to exert a paracrine and/or autocrine role within adipose tissue. TNFalpha affects many aspects of adipocyte function, from adipocyte development to lipid metabolism. Bringing together all of these diverse actions, TNFalpha appears to play a general role in reducing adipose tissue mass. Dysregulation of TNFalpha production and/or action could be one facet in the development of cachexia and obesity, as well as associated metabolic disorders such as insulin resistance. PMID- 12773115 TI - Adenosine signalling pathways in the pituitary gland: one ligand, multiple receptors. AB - Adenosine receptors are widely distributed in most species and mediate a diverse range of physiological and pathological effects. Although adenosine receptors have been identified in the pituitary gland, the distribution of the individual subtypes (A(1), A(2A), A(2B), A(3)) has not been well defined. Furthermore, the effects of adenosine on pituitary trophic activity and function are not well established despite good evidence for growth- and immune-modulating properties of the nucleoside elsewhere. Recent advances have provided a more detailed description of adenosine receptor distribution and function in the anterior pituitary and this commentary reviews these observations and highlights some of the possible implications in relation to the control of the hypothalamic pituitary-adrenal axis and the regulation of inflammation and pituitary cell growth. PMID- 12773116 TI - Proopiomelanocortin gene expression and DNA methylation: implications for Cushing's syndrome and beyond. AB - Proopiomelanocortin gene (POMC) is recognised as playing an important role in the regulation of the hypothalamo-pituitary-adrenal axis, adrenal development and obesity. POMC is activated in ACTH-dependent Cushing's syndrome. The syndrome may occur when the highly tIssue-specific 5' promoter of human POMC is activated in pituitary and non-pituitary sites. Whilst the factors involved in transcription in the corticotrophs of the anterior pituitary gland are becoming well delineated, the mechanism of activation in non-pituitary sites is not fully understood. This promoter is embedded within a defined CpG island, and, in contrast to somatically expressed CpG island promoters reported to date, is methylated in normal non-expressing tIssues, but is specifically unmethylated in expressing tIssues, tumours and the POMC-expressing DMS-79 small-cell lung cancer cell line. Methylation in vitro is sufficient for silencing of expression. In particular, methylation near the response element for the tIssue-specific POMC activator PTX1, diminishes POMC expression. Sites outside the PTX1 response element may be important for binding, and this may have implications for pituitary development. DMS-79 cells lack POMC-demethylating activity, implying that the methylation and expression patterns are likely to be set early or prior to neoplastic transformation, and that targeted de novo methylation might be a potential therapeutic strategy. It is conceivable that in POMC neurons of the hypothalamus the POMC promoter is subject to a variable density of methylation with clear implications for the signalling of satiety and obesity. PMID- 12773117 TI - Corticotrophin-releasing activity of desmopressin in Cushing's disease: lack of correlation between in vivo and in vitro responsiveness. AB - Desmopressin (DDAVP), an arginine vasopressin analogue, markedly stimulates ACTH secretion in patients with Cushing's disease, in contrast to its minimal effect in normal subjects. However, little is known about the mechanisms underlying this action and it appeared to be of interest to evaluate the effect of DDAVP on ACTH secreting pituitary adenomas in vitro, in comparison with its effect in the same patients in vivo. Pituitary adenomas from 14 patients with Cushing's disease were incubated with DDAVP, corticotrophin-releasing hormone (CRH) and DDAVP together with vasopressin receptor antagonists or CRH. Incubation with DDAVP induced a modest dose-dependent increase in ACTH concentrations which appeared maximal at 10 nM. CRH stimulated ACTH to a greater extent compared with DDAVP and potentiated the effect of DDAVP alone. The DDAVP-induced ACTH increase appeared blunted by vasopressin V(2) and V(3) receptor antagonists. V(3) receptor gene expression was detected by RT-PCR in all adenoma samples except for two which were not responsive to DDAVP in vitro but responsive to the peptide in vivo. Surprisingly, no difference in the in vitro ACTH secretory response was observed between in vivo DDAVP-responsive (ACTH peak>150% baseline) and -unresponsive (ACTH peak<120% baseline) patients, suggesting that the pituitary adenoma is not the sole mediator of the ACTH-releasing effect of DDAVP. In conclusion, the marked stimulatory effect of DDAVP observed in patients with Cushing's disease appears to be mainly dependent on an extrapituitary action, possibly the inhibition of a corticotrophin release-inhibitory factor. PMID- 12773118 TI - Short-term glucocorticoid treatment of prepubertal mice decreases growth and IGF I expression in the growth plate. AB - The insulin-like growth factor (IGF) system is an important mediator of postnatal longitudinal growth, and the growth inhibiting effects of glucocorticoid (GC) treatment are suggested to be due to impaired action of the IGF system. However, the precise changes of the IGFs and the IGF-binding proteins (IGFBPs) in the growth plate, occurring upon short-term GC treatment have not been characterized. Prepubertal mice treated daily with dexamethasone (DXM) for 7 days, showed significant growth inhibition of total body length and weight and weight of the liver, thymus and spleen, whereas the weight of the kidneys was not affected. Analysis of the tibial growth plate showed that the total growth plate width significantly decreased to 84.5% of control values, caused by a significant decrease in the proliferative zone. The number of proliferating cell nuclear antigen (PCNA)-positive chondrocytes in the proliferative zone decreased significantly (to 40%) and TUNEL staining showed a significant 1.6-fold increase in apoptotic hypertrophic chondrocytes. In the growth plates, both IGF-I and IGF II, as well as IGFBP-2 mRNAs were detected, mainly in the proliferative and prehypertrophic zones. DXM treatment significantly decreased the number of chondrocytes expressing IGF-I, whereas the number of chondrocytes expressing IGF II and IGFBP-2 were not affected. The decrease in IGF-I expression in the growth plate indicates that GC treatment affects IGF-I at the local level of the growth plate, which could contribute to the GC-induced growth retardation. PMID- 12773119 TI - Engineered IGF-I expression induces glandular enlargement in the murine prostate. AB - IGF-I has been implicated as a factor that may predispose one to prostate cancer and to benign prostatic hypertrophy (BPH). We established murine IGF-I transgenic mice under the control of rat probasin promoter and analysed the histology of the murine IGF-I-overexpressing prostate. Immunohistochemically, IGF-I was expressed in prostatic epithelial cells or basement membranes of the ventral, dorsal and lateral lobes in a line of IGF-I transgenic mice, but not in their control littermates. The anterior lobe did not express IGF-I. IGF-binding protein-3 (IGFBP-3), inhibitory to the mitogenic action of IGF-I, was detected in epithelial cells of prostatic ventral lobes, but not in those of the dorsal, lateral or anterior lobes of IGF-I transgenic mice. In controls, IGFBP-3 was not detected in epithelial cells of any prostatic lobe. Macroscopic prostatic size and the appearance of IGF-I transgenic mice were comparable with those of their control littermates of the same age. With a computed morphometric analysis, epithelial glands and intraglandular lumens in the prostatic lobes except the ventral lobe were smaller at 17 Months of age than at 14 Months both in IGF-I transgenic mice and controls. Glands and intraglandular lumens in the ventral prostatic lobes of IGF-I transgenic mice expressing more IGF-I protein in the prostate than controls were dense and enlarged similar to cysts compared with those of non-transgenic littermates without showing epithelial growth. Glands and lumens in the dorsal and lateral lobes of the IGF-I transgenic mice were also larger than controls at 14 and/or 17 Months of age. Glands in the anterior prostatic lobe of the IGF-I transgenic mice were not morphologically or morphometrically different from those of non-transgenic littermates. In conclusion, IGF-I transgenic mice under the control of rat probasin promoter showed more dense and enlarged epithelial glands in their prostatic ventral, dorsal and lateral lobes. PMID- 12773120 TI - Insulin inhibition of the proteasome is dependent on degradation of insulin by insulin-degrading enzyme. AB - A consequence of insulin-dependent diabetes mellitus is the loss of lean muscle mass as a result of accelerated proteolysis by the proteasome. Insulin inhibition of proteasomal activity requires interaction with insulin-degrading enzyme (IDE), but it is unclear if proteasome inhibition is dependent merely on insulin-NIDE binding or if degradation of insulin by IDE is required. To test the hypothesis that degradation by IDE is required for proteasome inhibition, a panel of insulin analogues with variable susceptibility to degradation by IDE binding was used to assess effects on the proteasome. The analogues used were [Lys(B28), Pro(B29)] insulin (lispro), [Asp(B10)]-insulin (Asp(B10)) and [Glu(B4), Gln(B16), Phe(B17)] insulin (EQF). Lispro was as effective as insulin at inhibition of degradation of iodine-125 ((125)I)-labeled insulin, but Asp(B10) and EQF were somewhat more effective. All agents inhibited cross-linking of (125)I-insulin to IDE, suggesting that all were capable of IDE binding. In contrast, although insulin and lispro were readily degraded by IDE, Asp(B10) was degraded more slowly, and EQF degradation was undetectable. Both insulin and lispro inhibited the proteasome, but Asp(B10) was less effective, and EQF had little effect. In summary, despite effective IDE binding, EQF was poorly degraded by IDE, and was ineffective at proteasome inhibition. These data suggest that insulin inhibition of proteasome activity is dependent on degradation by IDE. The mechanism of proteasome inhibition may be the generation of inhibitory fragments of insulin, or by displacement of IDE from the proteasome. PMID- 12773121 TI - Glucose-dependent insulinotropic polypeptide and insulin-like immunoreactivity in saliva following sham-fed and swallowed meals. AB - Gastrointestinal peptides, including insulin, glucagon and glucose-dependent insulinotropic polypeptide (GIP) have previously been reported in salivary glands. Recent evidence has suggested they might influence postprandial macronutrient metabolism. This study therefore investigated and compared postprandial hormone concentrations in saliva and plasma to determine whether their secretion was influenced by oral food stimuli. In a within-subject randomised cross-over comparison of hormone concentrations in plasma and saliva following a mixed meal, 12 subjects were given two 1708 kJ mixed meals. On one occasion the meal was chewed and swallowed (swallowed meal), on the other it was chewed and expectorated (sham-fed meal). Salivary and plasma levels of immunoreactive insulin, GIP and glucagon-like peptide-1 (GLP-1), total protein, alpha-amylase, glucose and non-esterified fatty acid were measured before and for 90 min following the meals. Saliva total protein and alpha-amylase rose following both meals, indicating that the stimulus for salivary protein release is related to the presence of food in the mouth. GLP-1 was not detected in saliva. Fasting salivary insulin levels were lower in saliva than plasma (28+/-6 vs 40+/-25 pmol/l respectively). Both increased following the swallowed meal but the rise in saliva was slower and less marked than in plasma (peak levels 96+/-18 and 270+/ 66 pmol/l for saliva and plasma respectively, P<0.01). Both were unchanged following the sham-fed meal. GIP was detected in saliva. Fasting GIP levels were significantly higher in saliva than plasma (183+/-23 compared with 20+/-7 pmol/l, P<0.01). They decreased in saliva following both swallowed and sham-fed meals to nadirs of 117+/-17 and 71+/-12 pmol/l respectively, but rose following the swallowed meal to peak levels of 268+/-66 pmol/l. These findings are consistent with insulin in saliva being an ultrafiltrate of that circulating in blood, but GIP in saliva being the product of local salivary gland synthesis, whose secretion is influenced, directly or indirectly, by oral stimuli. The function of salivary GIP is unknown, but we speculate that it may play a role in the regulation of gastric acid secretion in the fasting state. PMID- 12773122 TI - Hedgehog signaling enhances core-binding factor a1 and receptor activator of nuclear factor-kappaB ligand (RANKL) gene expression in chondrocytes. AB - Hedgehog signaling is considered to play a crucial role in chondrogenesis by regulation through a network of cytokine actions, which is not fully understood. We examined the effect of hedgehog signaling on the expression of core-binding factor a1 (Cbfa1), a critical transcription factor for the development of bone and cartilage. Primary chondrocytes prepared from the costal cartilage of newborn mice were treated with N-terminal fragment of recombinant murine sonic hedgehog (rmShh-N). Northern blot analysis indicated that Cbfa1 mRNA expression levels in the chondrocyte cultures were elevated by the treatment with rmShh-N. rmShh-N treatment enhanced 1.8 kb Cbfa1 promoter activity in chondrocytes, suggesting the presence of transcriptional control. As Cbfa1-binding site(s) have been located in the promoter of the receptor activator of nuclear factor-kappaB (RANK) ligand (RANKL) gene, we also examined RANKL expression. rmShh-N treatment upregulated RANKL and RANK mRNA expression levels in chondrocytes. Interestingly, RANKL suppressed the hedgehog enhancement of alkaline phosphatase activity in chondrocytes, suggesting the presence of a link between these signaling molecules. We conclude that hedgehog signaling activates Cbfa1 gene expression through its promoter in chondrocytes, and also activates and interacts with RANKL to maintain cartilage development. PMID- 12773123 TI - Interleukin-6 (IL-6), IL-1, receptor activator of nuclear factor kappaB ligand (RANKL) and osteoprotegerin production by human osteoblastic cells: comparison of the effects of 17-beta oestradiol and raloxifene. AB - Oestrogen inhibits bone resorption, at least in part, by regulating the production of several cytokines, including interleukin-6 (IL-6), IL-1, receptor activator of nuclear factor kappaB ligand (RANKL) and osteoprotegerin (OPG) by cells of the osteoblastic lineage. The selective oestrogen receptor modulator raloxifene (RAL) acts on bone in a similar manner to oestrogen, although the mechanisms of action of RAL on osteoblasts still remain unclear. We investigated and compared the effects of 17-beta oestradiol (E(2)) and RAL on the regulation of IL-6, IL-1, RANKL and OPG in vitro in primary human osteoblastic (HOB) cells and in an immortalised clonal human bone marrow stromal cell line (HCC1) with osteoblastic characteristics. We tested E(2) and RAL at concentrations ranging from 10(-12) to 10(-6) M. IL-6, IL-1alpha and IL-1beta, OPG and RANKL were measured by ELISA. RANKL and OPG mRNA steady state level was assessed by quantitative PCR analysis. Both E(2) and RAL led to a significant reduction in IL 6 production in the HOB cells, although the effect was more marked with E(2) (P<0.05). IL-1alpha and IL-1beta also decreased significantly following treatment with E(2) and RAL in the HCC1 cells (E(2) (10(-8), 10(-7) and 10(-6) M), % reduction (means+/-S.E.M.) compared with vehicle-treated cells - IL-1alpha: 84+/ 7.4, 70.8+/-2.9*, 78.2+/-4.8*; IL-1beta: 79+/-10, 72.8+/-8.2*, 66.6+/-2.8*; RAL (10(-8), 10(-7) and 10(-6) M) - IL-1alpha: 72.4+/-5*, 79+/- 5.2*, 102+/-7.7; IL 1beta: 67.9+/-3.2*, 69+/-2.5*, 73.8+/- 6.2*; *P<0.05). OPG protein concentration decreased significantly in a dose-dependent manner following treatment with E(2) and RAL (% reduction E(2) (10(-8), 10(-7) and 10(-6) M) - HOB: 72.5+/-8.4*, 80+/ 6.7*, 62.8+/-8.9*; HCC1: 109+/-4, 98.8+/-6, 54.5+/-3.4*; RAL (10(-8), 10(-7) and 10(-6) M) - HOB: 81.5+/-5.5*, 62.7+/-7.4*, 55.2+/-10.9*; HCC1: 92.7+/-7.4, 67+/ 12.2*, 39+/-4.5*; *P<0.05). In the HCC1 cells, RANKL protein did not change significantly following E(2). In contrast, a significant reduction in RANKL was seen with RAL at 10(-7) and 10(-6) M (66+/-6.4% and 74+/-3% respectively). There was no change in OPG mRNA expression following E(2) or RAL in the HCC1 cells, although in the HOB cells we observed a significant reduction in OPG mRNA. RANKL mRNA decreased significantly in the HCC1 cells following RAL (10(-8), 10(-7)and 10(-6) M) treatment (% change from controls: 52+/-2*, 62+/-1*, 53+/-5.8*; *P<0.05). Similar results were seen in the HOB cells with RAL at 10(-6) M (RANKL mRNA: 72+/-5.5, P<0.05). In addition, there was a significant decrease in the RANKL/OPG ratio after RAL at 10(-6) M (HOB: 65.6+/-5*, HCC1: 56.9+/-20*; *P<0.05). RANKL/OPG ratio did not change significantly in the HCC1 cells following E(2). However, in contrast to RAL, we observed an increase in the RANKL/OPG ratio in the HOB cells following treatment with E(2). In conclusion, the study shows that RAL and E(2) have divergent cell-specific effects on the regulation of cytokines. The data also suggest that, in contrast to E(2), RAL may exert its anti-resorptive actions, at least in part, via the RANKL/OPG pathway. Further in vivo studies are required to confirm this. PMID- 12773124 TI - The Booroola mutation in sheep is associated with an alteration of the bone morphogenetic protein receptor-IB functionality. AB - The hyperprolificacy phenotype of Booroola ewes is due to the presence of the FecB(B) allele at the FecB locus, recently identified as a single amino acid substitution (Q249R) in the bone morphogenetic protein (BMP) type-IB receptor (BMPR1B), and is associated with a more precocious differentiation of ovarian granulosa cells (GCs). To evaluate the consequences of the Booroola mutation on BMPR1B functions, the action of ligands of the transforming growth factor-beta (TGFbeta)/BMP family that act through (growth and differentiation factor-5, BMP 4) or independently of (activin A, TGFbeta-1) BMPR1B were studied on primary cultures of GCs from homozygous FecB(+) and FecB(B) ewes. All the tested TGFbeta/BMP family ligands inhibited progesterone secretion by FecB(+) GCs. Those inhibitory effects were lower for GCs from preovulatory (5-7 mm diameter) than from small antral follicles (1-3 mm diameter). The presence of the Booroola mutation was associated with a 3- to 4-fold (P<0.001) decreased responsiveness of GCs from FecB(B) compared with FecB(+) small follicles to the action of BMPR1B ligands. In contrast, TGFbeta-1 and activin A had similar inhibitory effects on progesterone secretion by GCs from FecB(+) and FecB(B) small follicles. No difference between genotypes was observed with GCs from preovulatory follicles. In transfection experiments with HEK-293 cells, co-expression of FecB(+) BMPR1B and BMPR2 resulted in a 2.6-fold (P<0.01) induction of the activity of a BMP specific luciferase reporter construct by BMP-4. Interestingly, no response to BMP-4 was observed when cells were transfected with the FecB(B) form of the BMPR1B receptor. Overall, these data strongly suggest that the Q249R mutation is associated with a specific alteration of BMPR1B signaling in hyperprolific Booroola ewes. PMID- 12773125 TI - Expression of 11beta-hydroxysteroid dehydrogenases in bovine follicle and corpus luteum. AB - In glucocorticoid target organs, local concentrations of active glucocorticoid are determined by the relative expression of two 11beta-hydroxysteroid dehydrogenases (HSDs): bi-directional 11beta-HSD type1 (11HSD1) that mainly activates cortisone to cortisol, and dehydrogenase 11beta-HSD type2 (11HSD2) that inactivates cortisol to cortisone. In this study, we examined the expression of mRNA encoding these two 11beta-HSDs in bovine granulosa cells harvested from preovulatory follicles and corpora lutea (CL). Ovaries were obtained from Holstein cows at a local slaughterhouse. Follicles larger than 10 mm in diameter and CL were dissected and follicular fluid and granulosa cells were taken. Corpora lutea were weighed and their stages were morphologically assessed (stage I, days 1-4; stage II, days 5-10; stage III, days 11-17; stage IV, days 8-20). Follicles were classified into four groups according to their hormonal status (oestradiol (E(2)): progesterone (P(4))>1: oestrogen active; E(2):P(4)<1: oestrogen inactive) and stage of the oestrous cycle (luteal or follicular phase). Total RNA was extracted with phenol-chloroform and subjected to a semi quantitative RT-PCR for 11HSD1, 11HSD2 and beta-actin. Concentrations of steroids in follicular fluid were determined by an enzyme immunoassay. In granulosa cells, only 11HSD1 mRNA was detected. There was a negative correlation between the expression of 11HSD1 and the concentration of cortisol in follicular fluid (P<0.05), indicating 11HSD1 may act as a dehydrogenase in the bovine follicle. Both types of 11beta-HSDs were expressed in CL. The levels of mRNA for both isozymes were high in stage I and II, and were decreased in stage III CL. In stage IV CL, the expression of 11HSD2 but not 11HSD1 mRNA increased. These results indicate that the bovine granulosa cells and CL express 11HSD1 and 11HSD2, and they may play an important physiological role in the bovine ovary through modulating the local glucocorticoid environment. PMID- 12773126 TI - Prolactin release during the estradiol-induced LH surge in ewes: modulation by progesterone but no evidence for prolactin-releasing peptide involvement. AB - An estradiol-induced prolactin surge accompanies the LH surge in several species, including sheep. However, the neural mechanisms underlying this surge remain poorly understood. A first study on estradiol- and progesterone-treated ovariectomized ewes examined whether the prolactin surge, like the LH surge, is sensitive to progesterone. Our data clearly showed that the estradiol-induced prolactin surge in the ewe is blocked by continuous exposure to progesterone and, importantly, that this blockade is overcome by pretreatment with the progesterone receptor antagonist, RU486. In a second study, we established that the generation of the prolactin surge is not dependent on the co-secretion of a prolactin releasing peptide in the hypophyseal portal blood or cerebrospinal fluid. The neuronal pathways targeted by estradiol and progesterone to modulate prolactin secretion at the time of the LH surge remain to be identified. Importantly, it has not been established whether there is any overlap in the neuronal systems generating the gonadotropin-releasing hormone and prolactin surges. PMID- 12773127 TI - Distribution and changes in amounts of the androgen receptor in the pig uterus during the estrous cycle, early pregnancy and after treatment with sex steroids. AB - Two experiments were performed to examine the expression of the androgen receptor (AR) gene in the pig uterus. In experiment 1, immunohistochemistry (IHC) was used to determine the distribution of the AR in uterine tIssue of pigs when collected at the first day of estrus (day 0) and the mid-luteal phase (day 12) of the estrous cycle, or early pregnancy (day 12, n=4 gilts per group). In experiment 2, AR immunostaining and AR mRNA in uterine tIssue were compared among ovariectomized gilts (n=4 per group) following treatment for 4 days with daily injections of: (1) progesterone (2 mg/kg bodyweight (BW)), (2) estradiol-17beta (E(2,) 2 micro g/kg BW), (3) E(2) plus progesterone (same dosages as 1 and 2 combined), (4) 5alpha-dihydrotestosterone (DHT, 7 micro g/kg BW), or (5) vehicle (corn oil). Data were analyzed using ANOVA. In experiment 1, nuclear staining for AR in luminal and glandular epithelia was strong and did not differ in intensity between the two locations. Immunostaining of AR in the myometrium was less (P<0.001) intense than in the luminal and glandular epithelia. Nuclei of stromal cells contained AR immunostaining that varied in intensity from strong (mainly in subepithelial stroma) to weak or no staining. Stages of the estrous cycle or early pregnancy did not influence AR immunostaining in the endometrial epithelia and myometrium. In experiment 2, immunostaining of AR in glandular and luminal epithelia and myometrium of ovariectomized gilts treated with vehicle or DHT was less (P<0.05) than in gilts treated with E(2), progesterone, or E(2) plus progesterone. Immunostaining of AR did not differ between ovariectomized gilts treated with vehicle or DHT, or between gilts treated with E(2), progesterone, or E(2) plus progesterone. In both experiments, intensity of AR immunostaining was greater in glandular epithelium located at the adluminal region compared with glandular epithelium located at the basal region of the endometrium. Competitive reverse-transcription PCR (RT-PCR) indicated a stimulatory effect (P<0.01) of E(2) on amounts of AR mRNA in whole endometrium. This increase in AR mRNA after E(2) treatment was not detected when E(2) was combined with progesterone. Endometrial AR mRNA was not influenced by DHT or progesterone relative to vehicle treated gilts. In conclusion, immunoreactive AR is mainly present in luminal and glandular epithelia of the pig uterus and to a lesser extent in the myometrium, and does not change significantly during the estrous cycle or early pregnancy. Expression of the AR gene in the pig endometrium and myometrium appears to be regulated by E(2) and progesterone. PMID- 12773128 TI - Methimazole-induced hypothyroidism in rats ameliorates oxidative injury in experimental colitis. AB - Depression of metabolism by hypothyroidism decreases oxidant production and thus protects tIssues against oxidant damage. Moreover, it is well-known that abnormal gut motility is a common manifestation in hypo/hyperthyroidism. In this study, we aimed to investigate the putative beneficial effects of methimazole on oxidative injury and dysmotility in a rat colitis model. Methimazole (0.04%) was administered in drinking water starting 15 days prior to induction of colitis. Colitis was induced by intracolonic administration of trinitrobenzene sulfonic acid (30 mg/ml; 0.8 ml) in ethanol. Six days after the induction of colitis, the fecal output was measured and used as an index for colonic motility. All rats were decapitated on the seventh day. The distal colon was weighed and the mucosal lesions were scored. Colonic lipid peroxidation (LP) and glutathione (GSH) measurements were performed. The macroscopic score, the colonic wet weight and LP values of the euthyroid colitis group were found to be higher than those of the control group (P<0.05-0.001). All these parameters were reduced in the methimazole-treated colitis group (P<0.01-0.001). The decrease in colonic GSH levels in the colitis group was completely abolished in the methimazole-treated colitis rats (P<0.01). Induction of colitis increased the average fecal output compared with the control group (P<0.05) and methimazole in the colitis group exaggerated the fecal output (P<0.001). In conclusion, methimazole reduces colonic oxidative injury probably due to hypometabolism, which is associated with a decrease in the production of reactive oxygen intermediates and an increase in the response of antioxidant systems. PMID- 12773129 TI - Outcome in patients with differentiated thyroid cancer with negative diagnostic whole-body scanning and detectable stimulated thyroglobulin. AB - BACKGROUND: Management of patients with differentiated thyroid carcinoma with negative diagnostic radioiodide scanning and increased serum thyroglobulin (Tg) concentrations is a widely debated problem. High-dose iodine-131 treatment of patients who have a negative (131)I diagnostic whole-body scan (WBS) is advocated. However, the therapeutic benefit of this "blind" treatment is not clear. OBJECTIVE: To investigate the course of serum Tg during thyroid hormone suppression therapy (Tg-on) and clinical outcome in patients with negative diagnostic (131)I scanning and increased serum Tg concentrations during thyroid hormone withdrawal (Tg-off), after treatment with high-dose (131)I. DESIGN: Retrospective single-center study. METHODS: Fifty-six patients were treated with a blind therapeutic dose of 150 mCi (131)I. Median follow-up from this treatment until the end of observation was 4.2 Years (range 0.5-13.5 Years). RESULTS: The post-treatment WBS revealed (131)I uptake in 28 patients, but none in the remaining 28 patients. In this study the Tg-on values did not change after treatment in either the positive or the negative post-treatment WBS group. During follow-up, 18 of the 28 patients with a positive post-treatment WBS achieved complete remission, compared with 10 of the 28 patients with a negative post treatment WBS. Nine patients in the negative group died, but no patients died in the positive post-treatment group (P=0.001). CONCLUSIONS: High-dose iodine treatment in diagnostically negative patients who have a negative post-treatment scan seems to confer no additional value for tumor reduction and survival. In patients with a positive post-treatment scan, high-dose iodine treatment can be used as a diagnostic tool to identify tumor location, and a therapeutic effect may be present in individual cases. PMID- 12773130 TI - Influence of renal function on clinico-pathological features of primary hyperparathyroidism. AB - OBJECTIVE: Disturbed renal function may play an important role in the clinico pathological presentation of primary hyperparathyroidism (pHPT). We studied the influence of renal function on the clinico-pathological characteristics of 141 patients (123 women and 18 men) with surgically proven pHPT. METHODS: The 141 patients were assigned to one of two groups based on creatinine clearance (C(cr)) level: a renal insufficiency group (n=37) in which C(cr) of patients was <70 ml/min and a normal renal function group (n=104) in which C(cr) was > or =70 ml/min. Clinical presentation and biochemical indices were evaluated and compared between the two groups. RESULTS: Age, and frequency of hypertension and of diabetes mellitus were significantly (P<0.001, P<0.05 and P<0.05 respectively) higher in the renal insufficiency group than in the normal renal function group. Serum levels of calcium, intact parathyroid hormone and bone Gla protein were significantly (P<0.05) higher and the excised parathyroid weighed significantly more (P<0.05) in the renal insufficiency group than in the normal renal function group; however, serum 1,25-dihydroxyvitamin D (1,25(OH)(2)D) and 24 h urinary calcium excretion were significantly (P<0.001 and P<0.05 respectively) lower in the former than in the latter group. There was a significant inverse correlation between C(cr) level and serum calcium (r=0.315, P<0.001) and a significant positive correlation between C(cr) level, 1,25(OH)(2)D (r=0.315, P<0.001), and 24 h calcium excretion (r=0.458, P<0.0001). CONCLUSIONS: Clinico-pathological features of pHPT were notably influenced by even moderate renal insufficiency. Urinary calcium excretion decreased according to the decrease in glomerular filtration rate. Therefore, endocrinologists need to appraise urinary calcium excretion and renal function of pHPT patients when considering surgery or in discriminating familial hypocalciuric hypercalcemia. PMID- 12773131 TI - Calcium-sensing receptor gene polymorphism is not associated with bone mineral density in Italian postmenopausal women. AB - OBJECTIVE: Calcium-sensing receptor (CaR) is a candidate gene for osteoporosis susceptibility. Several CaR polymorphisms have been identified and an association between the A986S genotype and serum calcium levels has been found in Canadian postmenopausal women. We investigated whether the presence of 986S allele was associated with bone mineral density (BMD) and osteoporotic fractures. DESIGN: The study group consisted of 164 Italian postmenopausal women without fragility fracture (Fx(-)) and 55 women with fracture (Fx(+)). METHODS: A fragment of exon 7 of CaR gene containing three polymorphisms (A986S, R990G and Q1011E) was amplified by PCR and sequenced. Anthropometric characteristics and BMD were evaluated. RESULTS: The A986S polymorphism was the most commonly observed (27.9%), whereas the other two CaR polymorphisms, R990G and Q1011E, occurred in a minority of cases (8.8 and 5.5% respectively). There was no significant difference in the frequency distribution of any CaR allele between Fx(-) and Fx(+) patients. Body mass index was found to predict BMD at the lumbar spine and femoral neck. The A986S polymorphism and Years since menopause were not independent predictors of BMD at any site. As far as fracture occurrence, there was no statistically significant difference in the prevalence of fractures between women carrying or not carrying the 986S allele. CONCLUSIONS: Our data do not support a role of A986S CaR polymorphism in BMD and in the prevalence of fragility fractures in Italian postmenopausal women. PMID- 12773132 TI - Severe hyponatremia due to hypopituitarism with adrenal insufficiency: report on 28 cases. AB - OBJECTIVE: Severe hyponatremia due to hypopituitarism and adrenal insufficiency can be life-threatening, and treatment with glucocorticoids is very effective once the diagnosis of the underlying disorder has been made. In our experience, the diagnosis of hypopituitarism in hyponatremic patients is often overlooked. METHODS: In a retrospective study we screened the files of 185 patients with severe hyponatremia (<130 mmol/l) that had been seen in one endocrinological unit of a university hospital between 1981 and 2001 in order to describe the clinical spectrum of patients with hyponatremia and hypopituitarism including secondary adrenal insufficiency. RESULTS: In 139 cases it was possible to clearly ascribe the patients to the pathophysiological groups of (i) primary sodium deficiency, (ii) edematous disorders, and (iii) normovolemic disorders including the "syndrome of inappropriate secretion of antidiuretic hormone" (SIADH). Twenty eight patients with severe "normovolemic hyponatremia" (serum sodium: 116+/-7 mmol/l, mean+/-s.d.) had hypopituitarism and secondary adrenal insufficiency as shown by basal cortisol measurements and dynamic tests of adrenal function. In 25 cases of this group hypopituitarism (mostly due to empty sella, Sheehan's syndrome and pituitary tumors) had not been recognized previously, and in 12 cases recurrent hyponatremia during previous hospital admissions (up to four times) could be documented. The mean age of these patients (21 women, seven men) was 68 Years. The most frequently occurring clinical signs were missing or scanty pubic and axillary hair, pale and doughy skin, and small testicles in the men. Frequent symptoms like nausea and vomiting, confusion, disorientation, somnolence or coma were similar to those in 91 patients with SIADH. Basal serum cortisol levels in the acutely ill state ranged from 20 to 439 nmol/l (mean+/-s.d.: 157+/ 123), while in 30 other severely hyponatremic patients it ranged from 274 to 1732 nmol/l (732+/-351 nmol/l). In most patients with hyponatremic hypopituitarism, plasma antidiuretic hormone levels were inappropriately high, probably due to a failure of endogenous cortisol to suppress the hormone in a stressful situation. All patients recovered after low-dose hydrocortisone substitution. Most patients had other pituitary hormone deficiencies and were appropriately substituted subsequently. CONCLUSIONS: Hypopituitarism including secondary adrenal insufficiency seems to be a frequently overlooked cause of severe hyponatremia. A high level of suspicion is the best way to recognize the underlying disorder. Treatment with hydrocortisone is very effective. PMID- 12773133 TI - The R271W mutant form of Pit-1 does not act as a dominant inhibitor of Pit-1 action to activate the promoters of GH and prolactin genes. AB - OBJECTIVE: Genetic abnormalities of the pituitary specific transcription factor, Pit-1, have been reported in several patients with GH, prolactin (PRL) and TSH deficiencies. The most common is a mutation altering an arginine to a tryptophan in codon 271 (R271W) in one allele of the Pit-1 gene. According to the previous in vitro expression study, R271W acted as a dominant negative inhibitor of the wild type to activate the GH promoter. However, healthy carriers with this mutation, who should be affected by the dominant negative effect of R271W, have also been reported. The aim of this study was to clarify in more detail the function of this mutant form of Pit-1. METHODS: Transcriptional activity of R271W for the expression of Pit-1-associated genes was investigated in COS7 cells with the aid of transient transfection assays. The 1.8 kb rat GH, 0.6 kb rat PRL or 1.9 kb rat PRL 5'-flanking regions were inserted upstream of the luciferase reporter gene and were used for functional analysis of R271W. Another reporter gene containing seven Pit-1 responsive elements was also used. The same experiments were also performed using JEG3 and CHO cells. RESULTS: We could not confirm the dominant negative effect of R271W on wild type Pit-1. Furthermore, our expression study revealed that R271W could activate the promoters of GH and PRL genes to levels similar to the wild type. CONCLUSION: Taken together with the evidence that phenotypically normal cases have been reported with this mutation, our results deny the relationship between R271W and combined pituitary hormone deficiency. PMID- 12773134 TI - Serum insulin-like growth factor binding protein-2 levels as an indicator of functional ability in elderly men. AB - BACKGROUND: In a cross-sectional study in 403 healthy, independently living elderly men (mean age 78 years), we determined which are the main physiological determinants of functional ability in the elderly, and which components of the somatotropic system contribute to the maintenance of functional ability. METHODS: Functional ability was assessed by the number of problems in activities of daily living and by a measure of physical performance. Other physical characteristics included leg extensor strength, bone mineral density of total body and proximal femur, and body composition, including lean mass and fat mass. Serum insulin-like growth factor (IGF)-I and its binding proteins (IGFBP) -1, -2 and -3 concentrations were all measured by RIA. RESULTS: Muscle strength was related to a lower degree of disability. Further, it was positively related to physical performance and bone mineral density (all P<0.001). Fat mass influenced activities of daily living and physical performance negatively and bone mineral density positively (all P<0.001). Serum concentrations of IGF-I and IGFBP-3 were not related to any of the physical characteristics. High serum IGFBP-2 concentrations were related to a higher degree of disability (P<0.001), a lower physical performance (P=0.006), muscle strength (P=0.002), bone mineral density of proximal femur (P=0.007), lean mass and fat mass (both P<0.001). Serum insulin and IGFBP-1 concentrations were independently, positively related to lean mass (P=0.003) and fat mass (P<0.001). CONCLUSIONS: In independently living elderly men, functional ability appears to be determined by muscle strength (positive) and fat mass (negative). Low serum IGFBP-2 concentrations are a powerful indicator for overall good physical functional status, probably inversely reflecting the integrated sum of nutrition and the biological effects of growth hormone, IGF-I and insulin. PMID- 12773135 TI - Acute hyperglycemia and activation of the beta-adrenergic system exhibit synergistic inhibitory actions on growth hormone (GH) releasing hormone-induced GH release. AB - OBJECTIVE: Acute hyperglycemia stimulates somatostatin (SRIH) release by the hypothalamus which, in turn, suppresses growth hormone (GH) secretion from the anterior pituitary gland. Although it has been suggested that the cholinergic pathway mediates glucose-induced SRIH release, other regulatory systems have not been examined. Therefore, we investigated whether blocking or activating the beta adrenergic pathway alters glucose-mediated inhibition of GH release. DESIGN AND METHODS: One set of experiments was performed with a beta-adrenergic antagonist, propranolol, and the other set with a beta-adrenergic agonist, isoproterenol. Each set of experiments was performed in ten healthy subjects and consisted of four tests. Test 1, a 100 microg GHRH bolus i.v. at 0 min; test 2, 100 g glucose orally at -30 min, followed by a 100 microg GHRH bolus at 0 min; test 3, after a 100 microg GHRH bolus i.v. at 0 min, a continuous infusion of propranolol (0.2 mg/kg) or isoproterenol (0.012 microg/kg) was administered between 0 and 120 min; test 4, after a 100 g glucose oral load at -30 min, and a 100 microg GHRH bolus i.v. at 0 min, a continuous infusion of propranolol (0.2 mg/kg) or isoproterenol (0.012 microg/kg) was administered between 0 and 120 min. Blood was drawn every 10 min from -30 min to 120 min to measure GH and glucose concentrations. RESULTS: Pretreatment with glucose significantly suppressed GHRH-induced GH secretion. Propranolol infusion significantly increased the GHRH-induced GH secretion, but it did not block glucose-induced suppression of GH secretion. Isoproterenol infusion alone significantly suppressed GHRH-induced GH secretion and augmented the inhibitory action of glucose on GH release. CONCLUSION: This study demonstrates that glucose-induced suppression of GHRH-stimulated GH release is independent of beta-adrenergic tone. Since previous data supports a role for SRIH in both glucose and beta-adrenergic suppression of GH release, the current results suggest that subsets of SRIH neurons are differentially responsive to these external cues. Therefore, a combined glucose and isoproterenol test may provide a useful assessment of hypothalamic somatostatinergic activity. PMID- 12773136 TI - Comparison of the impact of the I27L polymorphism of the hepatocyte nuclear factor-1alpha on estimated and measured beta cell indices. AB - OBJECTIVE: We investigated the impact of the I27L polymorphism of the hepatocyte nuclear factor-1alpha gene on measured and estimated beta cell indices. We also examined the conservation of this amino acid among different species. DESIGN AND METHODS: Estimated first and second phase insulin responses (1stPH(S) and 2ndPH(S)) were estimated from oral glucose tolerance tests in 78 glucose tolerant subjects. Among these subjects, first and second phase insulin responses (1stIR and 2ndIR) were measured in 60 subjects. The I27L genotypes were determined from genomic DNA. We also reviewed the published peptide sequence data on this polymorphism. RESULTS: The estimated beta cell indices correlated well with the measured indices. Although the impact of this polymorphism was noted in the measured indices (P<0.01 for 1stIR and P=0.04 for 2ndIR) from 60 subjects, the differences in the estimated indices were only noted in the extended sample set with 78 subjects (P=0.05 for 1stPH(S) and P=0.04 for 2ndPH(S)). This polymorphism occurs in the dimerization domain, which is completely conserved within human, rat, mouse and hamster. This amino acid is also conserved in chicken and zebrafish, but not in the frog. This conservation suggests a possible biological importance of this amino acid. CONCLUSIONS: By increasing the sample size, we demonstrated the role of the I27L polymorphism in the pathogenesis of type 2 diabetes by using estimated beta cell indices. The conservation among species suggests a possible biological importance of this amino acid. Analysis of the published data confirms a modest role of this polymorphism in type 2 diabetes. PMID- 12773137 TI - The effect of pegylated human recombinant leptin (PEG-OB) on neuroendocrine adaptations to semi-starvation in overweight men. AB - OBJECTIVE: Starvation induces a complex neuroendocrine response in humans thought to have evolved to defend against reduced energy intake. The drop in leptin levels observed during fasting has been implicated as a factor that triggers this adaptive response. To explore this hypothesis, we executed a randomized, double blind, placebo-controlled study to investigate whether elevated leptin levels using long-acting pegylated human recombinant leptin (PEG-OB) influenced the neuroendocrine responses to semi-starvation in human subjects. DESIGN: Twenty four overweight male subjects (mean+/-s.e.m.; 34.8+/-1.3 yrs; 28.8+/-0.5 kg/m(2)) were prescribed a very low energy diet (2.1 MJ/day) to induce a state of semi starvation for the next 46 days. In addition, all subjects received a weekly treatment of 80 mg PEG-OB or matching placebo. Hormone measurements were performed throughout the study period and included 5-h frequent hormone samplings and 24-h urine collections. RESULTS: Weekly subcutaneous administration of PEG-OB led to significant additional weight loss (2.8 kg) but it did not reverse the fasting-induced changes in the thyroid, corticotropic, somatotropic axes and sympathetic nervous system activity. However, after adjustment for weight loss, the drop in mean luteinizing hormone levels was attenuated in the PEG-OB group compared with the placebo group. CONCLUSIONS: These results suggest that a reduced level of leptin accompanying food restriction might be a component of the fasting-induced neuroendocrine inhibition of the human reproductive axis. PMID- 12773138 TI - Decreased plasma adiponectin concentrations in women with low-grade C-reactive protein elevation. AB - OBJECTIVE: Inflammation has been suggested as a risk factor for the development of atherosclerosis, while some components of metabolic syndrome X have been related to inflammatory markers. We hypothesized that adipocyte secreting protein, adiponectin and leptin, for which have been demonstrated an association with metabolic syndrome X and coronary artery disease, may be associated with inflammatory markers in nondiabetic humans. DESIGN AND METHODS: We measured high sensitivity C-reactive protein (hs-CRP), as an inflammatory marker, and adiponectin and leptin concentrations in 384 nondiabetic Japanese women (mean+/ s.e.m. age 53.6+/-0.8 Years, body mass index (BMI) 23.0+/-0.2 kg/m(2)) undergoing measurement of markers of metabolic syndrome X. RESULTS: The women who had a low grade hs-CRP elevation (>2.0 mg/l) were significantly older and had higher BMI, body fat mass (BFM), total cholesterol (TC), triglyceride (TG), atherogenic index (AI=(TC-HDLC)/HDLC), where HDLC is high-density lipoproten-cholesterol), fasting blood glucose and leptin concentrations before and after adjustment for BMI or BFM, while lower HDLC and adiponectin concentrations before and after adjustment compared with women with normal CRP levels (<0.5 mg/l). BMI, BFM, TG, AI and leptin before and after adjustment were found to be correlated with hs-CRP levels, while HDLC and adiponectin before and after adjustment were inversely correlated (all P<0.0001). hs-CRP was independently associated with white blood cell count, blood urea nitrogen and AI and inversely with adiponectin/BFM in the stepwise regression analysis model. CONCLUSIONS: These data demonstrate a significant decrease in plasma adiponectin in low-grade chronic inflammation, and suggest that there is an important linkage between inflammation/adipose tIssue/atherosclerosis. PMID- 12773139 TI - Adrenal function during coronary artery bypass grafting. AB - OBJECTIVE: To assess adrenal function in patients undergoing coronary artery bypass grafting (CABG) by means of the low-dose (1 microg) ACTH test, and to correlate the adrenal function with clinical outcome. METHODS: During a 5-Month period we prospectively included 45 patients undergoing elective CABG with cardiopulmonary bypass and without symptoms of endocrine disease. Low-dose (1 microg) ACTH tests were performed on the day before surgery (day -1), immediately after the operation (day 0), on the two subsequent days in the intensive care unit (day 1 and day 2), and on the day of discharge from the hospital. A number of clinical, hemodynamic and laboratory parameters were monitored throughout. RESULTS: On day -1, 75% of the study patients had normal stimulated plasma cortisol concentrations. Eleven patients (25%) had an impaired adrenal response to 1 microg ACTH. The stimulated plasma cortisol concentrations in patients who had an inadequate adrenal response on day -1 remained significantly reduced on day 1 (756+/-205 vs 949+/-259 nmol/l, P=0.03) (mean+/-s.d.), day 2 (644 (580-793) vs 885 (713-1087), P=0.03) (median (interquartile range)), and on the day of discharge (698+/-201 vs 854+/-186, P=0.05). In patients with a normal adrenal response in the preoperative setting peak cortisol concentrations were reached on day 1, in patients with a blunted adrenal response they were reached on day 2. There were significant correlations between the stimulated plasma cortisol concentrations and the blood loss (r=-0.50, P=0.002) and Volume balance (r=0.41, P=0.015). CONCLUSIONS: Occult (partial) adrenal insufficiency is common in patients undergoing CABG who are otherwise asymptomatic as regards endocrine disease. The adrenal function in these patients differs both in the magnitude of cortisol response to ACTH and in the time course, with significantly delayed peak cortisol concentrations. Adequate regulation of Volume balance and the amount of blood loss seem to correlate with adequacy of adrenal function. PMID- 12773141 TI - Antibiotic glycosyltransferases. AB - In the biosynthesis of several classes of antibiotics, sugars are attached to aglycone scaffolds by antibiotic-specific glycosyltransferases in the latter stages of the pathways. Two glycosylation pathways will be examined: the glycopeptide antibiotics of the vancomycin class and the aminocoumarin antibiotics of the novobiocin class. An oxidatively cross-linked heptapeptide scaffold is sequentially glucosylated and vancosaminylated by GtfE and GtfD, respectively, in vancomycin maturation, while in chloroeremomycin assembly the same heptapeptide is glucosylated by GtfB, then epivancosaminylated at two distinct sites by GtfA and GtfC. The specificity and mechanism of these glycosyltransferases will be discussed. In novobiocin biosynthesis, three enzymes (NovM, NovP and NovN) are thought to act sequentially to transfer an L-noviose moiety to the novobiocic acid aglycone (NovM), followed by 4'-hydroxyl methylation (NovP) and 3'-hydroxyl carbamoylation to produce the mature antibiotic structure, targeting the GyrB subunit of DNA gyrase. Initial characterization of NovM and NovP will be discussed. PMID- 12773142 TI - Oxygen activation in a copper-containing amine oxidase. AB - The process by which molecular oxygen is activated to enable it to function as an electron acceptor in biology is poorly understood. The quinoprotein copper containing amine oxidase (CuAO) catalyses the conversion of primary amines into aldehydes. As well as copper, the enzyme contains an organic cofactor, 2,4,5 trihydroxyphenylalanine quinone (TPQ). Following the formation of aldehyde, the enzyme is left as the two-electron reduced aminoquinol form. Reoxidation of the enzyme back to the resting state uses molecular oxygen, which is reduced to H(2)O(2) in the process, with the additional release of NH(3). To understand the structural basis of oxygen activation in Escherichia coli CuAO (ECAO), catalytically competent crystals were used to trap catalytic intermediates by exposing then to amine substrate and then freeze-trapping under aerobic and anaerobic conditions. Single-crystal visible microspectrophotometry was used to probe the oxidation state of the quinone in the intermediates, as TPQ exhibits a rich palette of colour changes during catalytic turnover. This review will focus on one of these structures, that of the rate-determining species in the crystal under steady-state conditions. This structure has revealed many details regarding oxygen activation in ECAO, including the site of dioxygen binding, and the proton transfer pathways involved in H(2)O(2) formation. PMID- 12773140 TI - Intralipid/heparin infusion suppresses serum leptin in humans. AB - BACKGROUND/AIM: Our previous studies showed that administration of dexamethasone plus food increased serum leptin levels 100% more than dexamethasone alone. We hypothesized that this increase in leptin from the meal could result directly from the provision of fuel metabolites rather than from the meal-induced rise in insulin. In the current study, we tested the effect of an i.v. lipid fuel source (Intralipid 20%/heparin) that would incur only a modest increase in insulin. This study was undertaken because the role of lipid in the regulation of human leptin levels has been controversial, with differing effects reported: stimulatory, inhibitory, or no effect at all. METHODS: In order to evaluate how lipids affect serum leptin in humans, we administered the following to seven lean, healthy, fasting subjects: (i) Intralipid 20% at 0.83 ml/kg.h plus heparin (800 IE/h) infused i.v. for 7 h (LIPID), (ii) LIPID with one initial pulse of insulin (0.09 U/kg) given s.c. (LIPID+INS), (iii) LIPID with dexamethasone (2 mg i.v. push) given at the start of the infusion (LIPID+DEX), and (iv) LIPID with insulin plus dexamethasone (LIPID+INS+DEX). Control trials in another 14 subjects matched hormonal conditions but lacked the LIPID infusion. Blood levels were collected over 8 h for determination of free fatty acids (FFA), glucose, insulin, and leptin under each experimental condition. RESULTS: Over the 420 min of LIPID infusion, FFA levels rose four-fold from 0.28+/-0.05 mmol/l to 0.99+/-0.05 mmol/l. Serum leptin levels were suppressed by 10-20% in the LIPID condition as compared with control (no LIPID) between 90 min (P=0.008) and 360 min (P=0.045). LIPID+DEX did not increase leptin. A pulse of insulin (INS) increased serum insulin levels to 49.9+/-6.1 U/ml at 90 min and increased serum leptin by 21.3+/ 6.6% at 480 min (P=0.054). LIPID decreased leptin in the face of this insulin induced increase (LIPID+INS), between 360 min (P=0.017) and 420 min (P=0.003), with a 23% suppressive effect at 420 min. LIPID+DEX elevated leptin levels by 112.5+/-35.8% at 480 min (P=0.037), however, the Intralipid/heparin infusion did not blunt the rise of leptin under these conditions. CONCLUSIONS: These data showed that Intralipid/heparin: (i) are not sufficient to trigger the effect of dexamethasone on leptin, (ii) have an acute inhibitory effect on both fasting and insulin-stimulated leptin levels, and (iii) that this inhibitory effect cannot reverse the strong stimulatory effect of dexamethasone and insulin on serum leptin. PMID- 12773143 TI - Electron transfer in human cytochrome P450 reductase. AB - Cytochrome P450 reductase (CPR) is a diflavin enzyme responsible for electron donation to mammalian cytochrome P450 enzymes in the endoplasmic reticulum. Dissection of the enzyme into functional domains and studies by site-directed mutagenesis have enabled detailed characterization of the mechanism of electron transfer using stopped-flow and equilibrium-perturbation methods, and redox potentiometry. These studies and the mechanism of electron transfer in CPR are reported herein. PMID- 12773144 TI - Calmodulin-dependent regulation of mammalian nitric oxide synthase. AB - The nitric oxide synthases are large, modular, dimeric enzymes composed of a reductase domain, which is related to cytochrome P450 reductase, and a structurally unique oxygenase domain containing a Cys-ligated haem. Both the neuronal and endothelial isoforms are activated by the reversible binding of calmodulin (CaM) at elevated intracellular Ca(2+) levels to produce NO as part of a number of cell signalling pathways. CaM binds to the linker region between the two domains and activates the enzyme by inducing intramolecular electron transfer. Protein-engineering experiments have shown that a series of unusual autoinhibitory inserts found only in the CaM-dependent NOS isoforms control both CaM binding and the structural rearrangement it induces. These lie in the reductase domain of the enzyme and include a 40-amino-acid autoinhibitory loop in the FMN-binding module, a 30-amino-acid extension to the C-terminus and the CaM binding site itself. The substrate (NADPH) also plays an important role in defining the CaM-dependence of the reductase domain by inducing a tight conformational lock in the absence of CaM. Both the substrate and the conformational lock appear to be released on CaM binding; the resultant domain mobility leads to activation. PMID- 12773145 TI - Cofactor processing in galactose oxidase. AB - Galactose oxidase (GO; EC 1.1.3.9) is a monomeric 68 kDa enzyme that contains a single copper and an amino acid-derived cofactor. The mechanism of this radical enzyme has been widely studied by structural, spectroscopic, kinetic and mutational approaches and there is a reasonable understanding of the catalytic mechanism and activation by oxidation to generate the radical cofactor that resides on Tyr-272, one of the copper ligands. Biogenesis of this cofactor involves the post-translational, autocatalytic formation of a thioether cross link between the active-site residues Cys-228 and Tyr-272. This process is closely linked to a peptide bond cleavage event that releases the N-terminal 17 amino-acid pro-peptide. We have shown using pro-enzyme purified in copper-free conditions that mature oxidized GO can be formed by an autocatalytic process upon addition of copper and oxygen. Structural comparison of pro-GO (GO with the prosequence present) with mature GO reveals overall structural similarity, but with some regions showing significant local differences in main chain position and some active-site-residue side chains differing significantly from their mature enzyme positions. These structural effects of the pro-peptide suggest that it may act as an intramolecular chaperone to provide an open active-site structure conducive to copper binding and chemistry associated with cofactor formation. Various models can be proposed to account for the formation of the thioether bond and oxidation to the radical state; however, the mechanism of prosequence cleavage remains unclear. PMID- 12773146 TI - The role of iron and 2-oxoglutarate oxygenases in signalling. AB - Sensing of ambient dioxygen levels and appropriate feedback mechanisms are essential processes for all multicellular organisms. In animals, moderate hypoxia causes an increase in the transcription levels of specific genes, including those encoding vascular endothelial growth factor and erythropoietin. The hypoxic response is mediated by hypoxia-inducible factor (HIF), an alphabeta heterodimeric transcription factor in which both the HIF subunits are members of the basic helix-loop-helix PAS (PER-ARNT-SIM) domain family. Under hypoxic conditions, levels of HIFalpha rise, allowing dimerization with HIFbeta and initiating transcriptional activation. Two types of dioxygen-dependent modification to HIFalpha have been identified, both of which inhibit the transcriptional response. Firstly, HIFalpha undergoes trans -4-hydroxylation at two conserved proline residues that enable its recognition by the von Hippel Lindau tumour-suppressor protein. Subsequent ubiquitinylation, mediated by an ubiquitin ligase complex, targets HIFalpha for degradation. Secondly, hydroxylation of an asparagine residue in the C-terminal transactivation domain of HIFalpha directly prevents its interaction with the co-activator p300. Hydroxylation of HIFalpha is catalysed by enzymes of the iron(II)- and 2 oxoglutarate-dependent dioxygenase family. In humans, three prolyl hydroxylase isoenzymes (PHD1-3) and an asparagine hydroxylase [factor inhibiting HIF (FIH)] have been identified. The role of 2-oxoglutarate oxygenases in the hypoxic and other signalling pathways is discussed. PMID- 12773147 TI - Haem-oxygen reactive intermediates: catalysis by the two-step. AB - The catalytic schemes of a variety of haem enzymes, including the P450 mono oxygenases, consist of a number of common reactive haem-oxygen adducts. The characterization of these intermediates by optical and EPR spectroscopies has reinforced the similarity of these intermediate states in a number of haem enzyme systems. Furthermore, the reactivity of these states in P450 and horseradish peroxidase, in which multiple potent oxidants are formed, provides a paradigm for many other haem enzymes. PMID- 12773148 TI - What can structure tell us about in vivo function? The case of aminoglycoside resistance genes. AB - Resistance to antibiotics used in the treatment of bacterial infections is an expanding clinical problem. Aminoglycosides, one of the oldest classes of natural product antibiotics, exert their bactericidal effect as the result of inhibiting bacterial protein synthesis by binding to the acceptor site of the 30 S ribosomal subunit. The most common mechanism of clinical resistance to aminoglycosides results from the expression of enzymes that covalently modify the aminoglycoside. We will discuss the enzymology and structure of two representative chromosomally encoded aminoglycoside N-acetyltransferases, Mycobacterium tuberculosis AAC(2') Ic and Salmonella enterica AAC(6')-Iy, and speculate about their possible physiological function and substrates. PMID- 12773149 TI - Mapping the conformational itinerary of beta-glycosidases by X-ray crystallography. AB - The conformational agenda harnessed by different glycosidases along the reaction pathway has been mapped by X-ray crystallography. The transition state(s) formed during the enzymic hydrolysis of glycosides features strong oxocarbenium-ion-like character involving delocalization across the C-1-O-5 bond. This demands planarity of C-5, O-5, C-1 and C-2 at or near the transition state. It is widely, but incorrectly, assumed that the transition state must be (4)H(3) (half-chair). The transition-state geometry is equally well supported, for pyranosides, by both the (4)H(3) and (3)H(4) half-chair and (2,5)B and B(2,5) boat conformations. A number of retaining beta-glycosidases acting on gluco -configured substrates have been trapped in Michaelis and covalent intermediate complexes in (1)S(3) (skew boat) and (4)C(1) (chair) conformations, respectively, pointing to a (4)H(3) conformed transition state. Such a (4)H(3) conformation is consistent with the tight binding of (4)E- (envelope) and (4)H(3)-conformed transition-state mimics to these enzymes and with the solution structures of compounds bearing an sp (2) hybridized anomeric centre. Recent work reveals a (1)S(5) Michaelis complex for beta-mannanases which, together with the (0)S(2) covalent intermediate, strongly implicates a B(2,5) transition state for beta-mannanases, again consistent with the solution structures of manno -configured compounds bearing an sp (2) anomeric centre. Other enzymes may use different strategies. Xylanases in family GH-11 reveal a covalent intermediate structure in a (2,5)B conformation which would also suggest a similarly shaped transition state, while (2)S(0)-conformed substrate mimics spanning the active centre of inverting cellulases from family GH-6 may also be indicative of a (2,5)B transition-state conformation. Work in other laboratories on both retaining and inverting alpha-mannosidases also suggests non-(4)H(3) transition states for these medically important enzymes. Three-dimensional structures of enzyme complexes should now be able to drive the design of transition-state mimics that are specific for given enzymes, as opposed to being generic or merely fortuitous. PMID- 12773150 TI - Molecular mechanism for inhibition of 3-hydroxy-3-methylglutaryl CoA (HMG-CoA) reductase by rosuvastatin. AB - The statins are inhibitors of 3-hydroxy-3-methylglutaryl CoA (HMG-CoA) reductase (HMG-CoAR), and are utilized to decrease levels of atherogenic lipoproteins in patients with, or who are at high risk of, cardiovascular disease. This study describes the inhibition of a recombinant, catalytic fragment of human HMG-CoAR by a new statin, rosuvastatin (CRESTOR(R)). Binding is reversible and involves an initial complex [inhibition constant involving the enzyme-inhibitor complex (E.I), K (i), approximately 1 nM], which undergoes a slow transition ( t ((1/2)) to reach steady state is 33-360 s) to give tighter association [steady-state inhibition constant involving E.I and the second E.I complex in a two-step mechanism (E.I*), K (i)*, approximately 0.1 nM]. At steady state, rosuvastatin is at least as potent as atorvastatin, cerivastatin and simvastatin. It is more potent than fluvastatin and pravastatin. For rosuvastatin, inhibition kinetics are competitive with respect to HMG-CoA and non-competitive when NADPH is varied. At 37 degrees C, binding is linked to a large favourable enthalpy change [Delta H degrees =-69.0 kJ/mol (-16.5 kcal/mol)] and a small entropic penalty [ T Delta S degrees =-9.6 kJ/mol (-2.3 kcal/mol)]. These characteristics, and the high affinity relative to that of 3 S -HMG-CoA ( K (d) approximately 6.6 microM), are discussed in relation to the crystal structures of complexes with HMG-CoAR. PMID- 12773151 TI - A structural perspective on the enzymes that convert dTDP-d-glucose into dTDP-l rhamnose. AB - Bacteria have a rich collection of biochemical pathways for the synthesis of complex metabolites. These conversions often involve chemical reactions that are hard to reproduce in the laboratory. An area of considerable interest is in the manipulation and synthesis of carbohydrates. In contrast with amino acids, carbohydrates are densely functionalized (each carbon atom is attached to at least one heteroatom) and this holds out the prospect of discovering novel enzyme mechanisms. The results from the study of the biosynthetic dTDP-L-rhamnose pathway are discussed. dTDP-L-rhamnose is a key intermediate in many pathogenic bacteria, as it is the donor for L-rhamnose, which is found in the cell wall of important human pathogens, such as Mycobacteria tuberculosis and Salmonella typhimurium. All four enzymes have been structurally characterized; in particular, the acquisition of structural data on substrate complexes was extremely useful. The structural data have guided site-directed-mutagenesis studies that have been used to test mechanistic hypotheses. The results shed light on three classes of enzyme mechanism: nucleotide condensation, short-chain dehydrogenase activity and epimerization. PMID- 12773152 TI - Structure and reactivity in the non-mevalonate pathway of isoprenoid biosynthesis. AB - The function, structure and mechanism of two Escherichia coli enzymes involved in the non-mevalonate route of isoprenoid biosynthesis, 2C-methyl-D-erythritol 4 phosphate cytidylyltransferase and 2C-methyl-D-erythritol 2,4-cyclodiphosphate synthase, are reviewed. Comparisons of each with enzymes from microbial pathogens highlight important conservation of sequence suggestive of similarities in secondary structure, subunit folds, quaternary structure and active sites. Since both enzymes are validated drug targets, the models provide templates for structure-based design of anti-microbial agents targeting a number of serious human diseases. PMID- 12773153 TI - Twists and turns: a tale of two shikimate-pathway enzymes. AB - We are studying two enzymes from the shikimate pathway, dehydroquinate synthase (DHQS) and 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS). Both enzymes have been the subject of numerous studies to elucidate their reaction mechanisms. Crystal structures of DHQS and EPSPS in the presence and absence of substrates, cofactors and/or inhibitors are now available. These structures reveal movements of domains, rearrangements of loops and changes in side-chain positions necessary for the formation of a catalytically competent active site. The potential for using complementary small-angle X-ray scattering (SAXS) studies to confirm the presence of these structural differences in solution has also been explored. Comparative analysis of crystal structures, in the presence and absence of ligands, has revealed structural features critical for substrate-binding and catalysis. We have also analysed these structures by generating GRID energy maps to detect favourable binding sites. The combination of X-ray crystallography, SAXS and computational techniques provides an enhanced analysis of structural features important for the function of these complex enzymes. PMID- 12773154 TI - Experiences with the shikimate-pathway enzymes as targets for rational drug design. AB - The background and current context of work on the shikimate-pathway enzymes as potential targets for anti-bacterial, anti-fungal and anti-parasitic drugs is reviewed. Recent work on the third enzyme of the pathway, dehydroquinase, which occurs in two structurally and mechanistically distinct forms, is used to illustrate the present state of studies into rational drug design. PMID- 12773155 TI - A two-faced molecule offers NO explanation: the proximal binding of nitric oxide to haem. AB - Cytochrome c ' (cyt c ') is found in the periplasmic space of denitrifying bacteria where it is thought to mediate the transfer of NO between the nitrogen cycle enzymes dissimilatory nitrite reductase and nitric oxide reductase. It contains a 5-coordinate (5c) His-ligated haem that shares spectroscopic and ligand-binding properties with the haem group in the sensory domain of soluble guanylate cyclase (sGC). The latter is an extremely important enzyme involved in the control of vasodilation and blood clotting. Curiously, the enzyme is activated up to 200-fold by the binding of NO to the haem, whereas the binding of CO gives rise to only a mild stimulation of activity. Through X-ray crystallography we have studied NO and CO binding to cyt c '. CO binds to the distal face to give a 6-coordinate (6c) adduct. By contrast, NO binding gives rise to a 5c adduct through the displacement of the proximal His, to give a novel and unexpected proximal binding mode for NO. These results are also supported by a range of spectroscopies. In the absence of a crystal structure for sGC we propose that cyt c ' provides a structural model for the haem domain of this enzyme and thereby helps to explain the differential effects of NO and CO on its activity. PMID- 12773156 TI - Engineering substrate recognition in catalysis by cytochrome P450cam. AB - We have a continuing interest in applying the current knowledge of cytochrome P450cam substrate recognition to engineer the enzyme for the biotransformation of unnatural substrates with the long-term aim of applications in the synthesis of fine chemicals and bioremediation of environmental contaminants. Comparisons of the structure of target substrates with that of camphor, the natural substrate, led to the design of active-site mutants with greatly enhanced activity for the oxidation of chlorinated benzenes and selectivity of (+)-alpha-pinene oxidation. The crystal structures of the F87W/Y96F/V247L mutant with 1,3,5-trichlorobenzene or (+)-alpha-pinene bound have revealed the enzyme-substrate contacts and provided insights into the activity and selectivity patterns. The structures have also provided a novel basis for further engineering of P450cam for increased activity in the oxidation of the highly inert pentachlorobenzene and hexachlorobenzene, and increased selectivity of (+)-alpha-pinene oxidation. PMID- 12773157 TI - Structural insights into the evolution of the pantothenate-biosynthesis pathway. AB - Pantothenate is synthesized in bacteria, fungi and plants, and as vitamin B5 is a dietary requirement in animals. The three-dimensional structures of the four Escherichia coli enzymes involved in the production of pantothenate have been determined. We describe the use of comparative analyses of the sequences and structures to identify distant homologues of the four enzymes in an attempt to understand the evolution of the pathway. We conclude that it is likely to have evolved via a patchwork mechanism, whereby the individual enzymes were recruited separately. PMID- 12773158 TI - United at last: the tuberous sclerosis complex gene products connect the phosphoinositide 3-kinase/Akt pathway to mammalian target of rapamycin (mTOR) signalling. AB - The molecular interplay between the phosphoinositide 3-kinase (PI3K) pathway and mammalian target of rapamycin (mTOR) signalling in the control of cell growth and proliferation has been the subject of much interest and debate amongst cell biologists. A recent escalation of research in this area has come from the discovery of the tuberous sclerosis complex gene products, tuberin and hamartin, as central regulators of mTOR activation. The PI3K effector Akt/protein kinase B has been found to directly phosphorylate tuberin and is thereby thought to activate mTOR through inhibition of the tuberin-hamartin complex. The many recent studies aimed at defining the molecular nature of this revamped PI3K/Akt/mTOR pathway are reviewed here. The collective data discussed have laid the groundwork for important new insights into the many cancers caused by aberrant PI3K activation and the clinically challenging tuberous sclerosis complex disease and have suggested a possible means of treatment for both. PMID- 12773159 TI - Aspects of tuberous sclerosis complex (TSC) protein function in the brain. AB - Tuberous sclerosis complex (TSC), an autosomal dominant disease caused by mutations in either TSC1 or TSC2, is characterized by the development of hamartomas in a variety of organs. Concordant with the tumour-suppressor model, loss of heterozygosity (LOH) is known to occur in these hamartomas at both TSC1 and TSC2 loci. LOH has been documented in renal angiomyolipomas, but loss of the wild-type allele in cortical tubers appears very uncommon. We analysed 24 hamartomas from 10 patients for second-hit mutations by several methods, and found no evidence for the inactivation of the second allele in many of the central nervous system (CNS) lesions, including tumours that appear to be clonally derived. We believe that somatic mutations in TSC1 and TSC2 resulting in the loss of wild-type alleles may not be necessary in some tumour types, and other mechanisms may contribute to tumorigenesis in this setting. We have shown that hamartin interacts with neurofilament light chain (NF-L) and could integrate the neuronal cytoskeleton through its direct interaction with NF-L and ERM (ezrin/radixin/moeisin) proteins. Our unpublished work further documents the binding of tuberin with Pam, a protein associated with c-Myc, which is enriched in brain. All these observations suggest that the tuberin-hamartin complex is likely to have distinct functions in the CNS. PMID- 12773160 TI - The tuberous sclerosis complex (TSC) pathway and mechanism of size control. AB - We have identified three groups of growth-constraint genes using mosaic genetic screens in Drosophila melanogaster, including PTEN (phosphatase and tensin homologue deleted on chromosome 10), and the tuberous sclerosis complex (TSC) genes, Tsc1 and Tsc2. Our studies show that all three groups of genes participate in mechanisms that regulate organ and organism size in animals. We propose that mechanisms of organ size control are critical targets for diseases, such as tumorigenesis, which require an increase in tissue size and total mass, and for evolutionary events that alter the size of organisms. Using genetic and biochemical methods, we have shown that Tsc1 and Tsc2 function in the insulin/phosphoinositide 3-kinase (PI3K)/Akt pathway. We have shown that Akt regulates the Tsc1-Tsc2 complex by directly phosphorylating Tsc2. We have shown further that S6 kinase (S6K) is a downstream component of the PI3K/Akt/TSC pathway and reduction of S6K activity can block TSC defects. Recent studies from many laboratories have now confirmed our findings in mice, rats and human patients, and have shown that drugs that antagonize S6K activities, such as rapamycin, diminish tumours in TSC-deficient mice and rats. Clinical trials based on these findings have begun. Given that other components of the pathway, such as PTEN, are also mutated in a large number of cancer patients and that these components regulate intracellular insulin signalling, therapeutics based on the knowledge of the pathway could have effects beyond the TSC patient population. PMID- 12773161 TI - Regulation of tuberous sclerosis complex (TSC) function by 14-3-3 proteins. AB - Tuberous sclerosis complex (TSC) is a genetic disorder characterized by seizures, mental disability, renal dysfunction and dermatological abnormalities. The disease is caused by inactivation of either hamartin or tuberin, the products of the TSC1 and TSC2 tumour-suppressor genes. Hamartin and tuberin form a complex and antagonise phosphoinositide 3-kinase/protein kinase B/target of rapamycin signal transduction by inhibiting p70 S6 kinase, an activator of translation, and activating 4E-binding protein 1, an inhibitor of translation initiation. Phosphorylation-dependent binding between tuberin and members of the 14-3-3 protein family indicates how the tuberin-hamartin complex may interact with upstream and downstream effectors, and suggests how phosphorylation-dependent regulation of the complex may be controlled. PMID- 12773162 TI - TSC1 and TSC2: genes that are mutated in the human genetic disorder tuberous sclerosis. AB - The tuberous sclerosis complex genes TSC1 and TSC2 were first identified by positional cloning strategies in the heritable human disorder tuberous sclerosis. They encode previously unknown proteins, termed hamartin and tuberin respectively, that form a functional complex. The phenotypic manifestations of tuberous sclerosis are extremely diverse and suggest normal roles for TSC1 and TSC2 in regulating the growth, proliferation, migration and differentiation of many cell types. Investigations of TSC1 and TSC2 in a number of model organisms and cell-culture systems have provided new insights into the mechanisms through which these roles are effected. Most promisingly, the hamartin-tuberin complex has been shown to function as a negtive regulator of the insulin receptor/phosphoinositide 3-kinase/S6 kinase pathway. Drugs that act to inhibit this pathway may have therapeutic potential for tuberous sclerosis and the related disorder lymphangioleiomyomatosis. PMID- 12773163 TI - Tuberous sclerosis complex (TSC) gene involvement in sporadic tumours. AB - In tuberous sclerosis patients, inactivation of the tuberous sclerosis complex tumour-suppressor genes TSC1 and TSC2 contributes to the development of a wide range of hamartomatous lesions. These patients do not, however, show an increased risk of the common adult solid cancers. Recent evidence that the TSC genes play a role in the phosphoinositide 3-kinase pathway, a pathway whose dysregulation is implicated in a wide range of human malignancies, raises the possibility that their inactivation could contribute to the development of some sporadic cancers. To date the only evidence for this comes from the finding of mutations of TSC1 in bladder cancer. The mutation spectrum of TSC1 in bladder cancer and functional evidence from TSC1 -gene-replacement studies in bladder tumour cells will be presented. The literature on genetic changes in several other sporadic epithelial cancers reveals relatively common deletions in the region of the TSC genes. In ovarian and gall bladder carcinoma and non-small-cell carcinoma of the lung, deletions in both 16p13 and 9q34 are found at significant frequency. Mutation analyses in such tumours are now merited. PMID- 12773164 TI - Similarity-based approaches to virtual screening. AB - Current similarity measures for virtual screening are based on the use of molecular fingerprints and the Tanimoto coefficient. This paper describes two ways in which one can increase the effectiveness of similarity-based virtual screening: using similarity coefficients other than the Tanimoto coefficient for the comparison of molecular fingerprints; and using a graph-theoretic similarity measure based on the largest substructure common to a pair of molecules. PMID- 12773165 TI - Targeting metabolic pathways in microbial pathogens: oxidative stress and anti folate drug resistance in trypanosomatids. AB - The large quantity of genomic, biochemical and metabolic data on microbial pathogens provides information that helps us to select biological problems of interest and to identify targets, metabolic pathways or constituent enzymes, for therapeutic intervention. One area of potential use in developing novel anti parasitic agents concerns the regulation of oxidative stress, and we have targeted the trypanothione peroxidase pathway in this respect. In order to characterize this pathway, we have determined crystal structures for each of its components, and are now studying enzyme-ligand complexes of the first enzyme, trypanothione reductase. Also with regard to trypanosomatids, a question that arose was: why do anti-folates not provide useful therapies? The enzyme pteridine reductase has been shown to contribute to anti-folate drug resistance, and we have determined the enzyme structure and mechanism to understand this aspect of drug resistance. PMID- 12773166 TI - In silico approaches to predicting drug metabolism, toxicology and beyond. AB - The discovery and optimization of new drug candidates is becoming increasingly reliant upon the combination of experimental and computational approaches related to drug metabolism, toxicology and general biopharmaceutical properties. With the considerable output of high-throughput assays for cytochrome-P450-mediated drug drug interactions, metabolic stability and assays for toxicology, we have orders of magnitude more data that will facilitate model building. A recursive partitioning model for human liver microsomal metabolic stability based on over 800 structurally diverse molecules was used to predict molecules with known log in vitro clearance data (Spearman's rho -0.64, P <0.0001). In addition, with solely published data, a quantitative structure-activity relationship for 66 inhibitors of the potassium channel human ether-a-gogo (hERG) that has been implicated in the failure of a number of recent drugs has been generated. This model has been validated with further published data for 25 molecules (Spearman's rho 0.83, P <0.0001). If continued value is to be realized from these types of computational models, there needs to be some applied research on their validation and optimization with new data. Some relatively simple approaches may have value when it comes to combining data from multiple models in order to improve and focus drug discovery on the molecules most likely to succeed. PMID- 12773167 TI - Arylamine N-acetyltransferases: a pharmacogenomic approach to drug metabolism and endogenous function. AB - The arylamine N-acetyltransferases (NATs) are a unique family of enzymes that catalyse the transfer of an acetyl group from acetyl-CoA to the terminal nitrogen of hydrazine and arylamine drugs and carcinogens. The NATs have been shown to be important in drug detoxification and carcinogen activation, with humans possessing two isoenzymes encoded by polymorphic genes. This polymorphism has pharmacogenetic implications, leading to different rates of inactivation of drugs, including the anti-tubercular agent isoniazid and the anti-hypertensive drug hydralazine. Mice provide a good model for human NAT, allowing genetic manipulation of expression to explore possible endogenous roles of these enzymes. The first three-dimensional NAT structure was resolved for NAT from Salmonella typhimurium, and subsequently the structure of NAT from Mycobacterium smegmatis has been elucidated. These identified a 'Cys-His-Asp' catalytic triad (conserved in all NATs), which is believed to be responsible for the activation of the active site cysteine residue. As more genomic data become available, NAT homologues continue to be found in prokaryotic species, many of which are pathogenic, including Mycobacterium tuberculosis. The discovery of NAT in M. tuberculosis is particularly significant, since this enzyme participates in inactivation of isoniazid in the bacterium, with implications for isoniazid resistance. Structural studies on NAT proteins and phenotypic analyses of organisms (both mice and prokaryotes) following genetic modifications of the nat genes are leading to an understanding of the potentially diverse roles of NAT in endogenous and xenobiotic metabolism. These studies have indicated that NAT, particularly in Mycobacteria, has the potential to be a drug target. Combinatorial chemical approaches, together with in silico structural studies, will allow for advances in the identification of NAT substrates and inhibitors, both as experimental tools and as potential drugs. PMID- 12773168 TI - Human carboxylesterase 1: from drug metabolism to drug discovery. AB - Human carboxylesterase 1 (hCE1) is a serine esterase involved in both drug metabolism and activation, as well as other biological processes. hCE1 catalyses the hydrolysis of heroin and cocaine, and the transesterification of cocaine in the presence of ethanol to the toxic metabolite cocaethylene. We have determined the crystal structures of hCE1 in complex with either the cocaine analogue homatropine or the heroin analogue naloxone. These are the first structures of a human carboxylesterase, and they provide details about narcotic metabolism in humans. hCE1's active site contains rigid and flexible pockets, explaining the enzyme's ability to act both specifically and promiscuously. hCE1 has also been reported to contain cholesteryl ester hydrolase, fatty acyl-CoA hydrolase and acyl-CoA:cholesterol acyltransferase activities, and thus appears to be involved in cholesterol metabolism. Since the enzyme may be useful as a treatment for cocaine overdose, and may afford protection against chemical weapons like Sarin, Soman and VX gas, hCE1 could serve as both a drug and a drug target. Selective hCE1 inhibitors targeted to several sites on the enzyme may also pave the way for novel clinical tools to manage cholesterol homoeostasis in humans. PMID- 12773169 TI - Cytochromes P450: novel drug targets in the war against multidrug-resistant Mycobacterium tuberculosis. AB - Novel drug strategies are desperately needed to combat the global threat posed by multidrug-resistant strains of Mycobacterium tuberculosis (Mtb). The genome sequence of Mtb has revealed an unprecedented number of cytochrome P450 enzymes in a prokaryote, suggesting fundamental physiological roles for many of these enzymes. Several azole drugs (known inhibitors of cytochromes P450) have been shown to have potent anti-mycobacterial activity, and the most effective azoles have extremely tight binding constants for one of the Mtb P450s (CYP121). The structure of CYP121 has been determined at atomic resolution, revealing novel features of P450 structure, including mixed haem conformations and putative proton-relay pathways from protein surface to haem iron. The structure provides both a platform for investigation of structure/mechanism of cytochrome P450, and for design of inhibitor molecules as novel anti-tubercular agents. PMID- 12773170 TI - Circular dichroism and synchrotron radiation circular dichroism spectroscopy: tools for drug discovery. AB - CD spectroscopy is an established and valuable technique for examining protein structure, dynamics and folding. Because of its ability to sensitively detect conformational changes, it has important potential for drug discovery, enabling screening for ligand and drug binding, and detection of potential candidates for new pharmaceuticals. The binding of the anti-tumour agent Taxol to the anti apoptosis protein Bcl-2 [Rodi, Janes, Sanganee, Holton, Wallace and Makowski (1999) J. Mol. Biol. 285, 197-204] and the binding of the anti-epileptic drug lamotrigine to voltage-gated sodium channels [Cronin, O'Reilly, Duclohier and Wallace (2003) J. Biol. Chem. 278, 10675-10682] are used as examples to show changes detectable by CD involving secondary structure, and are contrasted with the binding of the agonist carbamylcholine to acetylcholine receptors [Mielke and Wallace (1988) J. Biol. Chem. 263, 8177-8182], an example where binding does not involve a secondary structural change. Synchrotron radiation CD spectroscopy offers significant enhancements with respect to conventional CD spectroscopy, which will enable its usage for high-throughput screening and as a tool in 'chemical genomics' or 'reverse chemical genetics' strategies for ligand identification. The lower wavelength data available enable more detailed, sensitive and accurate detection, the higher light intensity permits much smaller amounts of both proteins and drug candidates to be used in the screening, and future technological developments in sample handling and detection should enable automated high-throughput screening to be performed. PMID- 12773171 TI - Nicotinic acetylcholine receptors: alpha-conotoxins as templates for rational drug design. AB - Nicotinic acetylcholine receptors (nAChRs) mediate the passage of potassium and sodium ions across synaptic membranes. Two classes of receptors exist: the neuromuscular nAChRs, which mediate signals between nerve and muscle cells, and the neuronal nAChRs, which are found throughout the nervous system. For treatment of diseases involving nAChRs, drugs must be designed with a high level of selectivity towards only one of these classes or subclasses (in the case of neuronal receptors). alpha-Conotoxins, small polypeptides isolated from the venoms of marine snails, represent molecules with just this type of selectivity, with specificity even towards certain subclasses of nAChRs. The availability of high-resolution crystal structures of alpha-conotoxins provides the opportunity to examine the structural features that orchestrate their preferential blocking action. In the present study of a neuromuscular- and a neuronal-specific alpha conotoxin, SI and EpI respectively, important and significant differences can be seen in the shapes of the molecules, which must reflect topological features of the different types of target receptor subunits. These then provide a template for computational docking studies with the homologous acetylcholine-binding protein, whose structure is known, so drug analogues of the naturally occurring toxins can be developed with the desired specificities. PMID- 12773172 TI - Role of adapters in Toll-like receptor signalling. AB - Toll-like receptors (TLRs) play a critical role in the detection of invading pathogens within the body and the subsequent immune response. Individual TLRs recognize distinct microbial components. The TLRs are a type 1 transmembrane receptor that possess an extracellular leucine-rich repeat domain and cytoplasmic domain homologous with that of the interleukin 1 receptor (IL-1R) family. Upon stimulation, TLR recruits the IL-1R-associated kinase (IRAK) via the adapter MyD88, ultimately leading to the activation of nuclear factor-kappaB. Cytokine production in response to all TLR ligands is completely abolished in MyD88 deficient cells, indicating that MyD88 is an essential signalling molecule shared among members of the IL-1R/Toll family. However, several novel adaptor molecules have recently been identified. Evidence is now accumulating showing that differential utilization of these adaptors may activate overlapping as well as distinct signalling pathways, and ultimately give rise to distinct biological effects exerted by individual TLR family members. PMID- 12773173 TI - The role of MyD88-like adapters in Toll-like receptor signal transduction. AB - Signal-transduction pathways activated by Toll-like receptors (TLRs) have been the subject of intense investigation because of the key role played by TLRs in the recognition and elimination of microbes. Signalling is initiated by a domain termed the Toll/interleukin-1 (IL-1) receptor (TIR) domain that occurs on the cytosolic face of TLRs. This recruits, via homotypic interactions, adapter proteins that contain TIR domains. Three such adapter proteins have been discovered to date, and have been named MyD88, Mal [MyD88 adapter-like; also known as TIRAP (TIR domain-containing adapter protein)] and Trif (TIR-domain containing adapter inducing interferon-beta). Differences are emerging between TLRs in terms of which adapter is recruited by which TLR. This may lead to specificities in TLR signalling, with pathways being triggered that are specific for the elimination of the invading microbe. However, signals that separate Mal from MyD88 have yet to emerge, although biochemical differences between the two proteins imply that each will have a specific function. PMID- 12773175 TI - Polymorphisms in Toll-like receptor 4 and the systemic inflammatory response syndrome. AB - The systemic inflammatory response syndrome (SIRS) is a major cause of morbidity and mortality, and is thought to be due to an over-amplification of an inflammatory response. The Toll-like receptor 4 ( TLR4 ) Asp-299-->Gly polymorphism has been shown to reduce lipopolysaccharide responsiveness. We examined whether this TLR4 polymorphism is associated with severity of SIRS. A trend was found between the minor allele and mortality in SIRS (odds ratio of 4.3; P =0.076), suggesting a role for TLR4 signalling in the severity of SIRS. PMID- 12773174 TI - Toll and Toll-like receptors in Drosophila. AB - The Drosophila Toll receptor controls the immune response to Gram-positive bacteria and fungi by activating a signalling pathway partially conserved throughout evolution. The Drosophila genome encodes eight additional Toll-related receptors, most of which appear to carry out developmental rather than immune functions. One exception may be Toll-9, which shares structural and functional similarities with mammalian TLRs. PMID- 12773176 TI - Toll-like receptor 9: modulation of recognition and cytokine induction by novel synthetic CpG DNAs. AB - Bacterial and synthetic DNA containing unmethylated 2'-deoxyribo(cytidine phosphate-guanosine) (CpG) dinucleotides in specific sequence contexts activate the vertebrate innate immune system. A molecular pattern recognition receptor, Toll-like receptor 9 (TLR9), recognizes CpG DNA and initiates the signalling cascade, although a direct interaction between CpG DNA and TLR9 has not been demonstrated yet. TLR9 in different species exhibits sequence specificity. Our extensive structure-immunostimulatory activity relationship studies showed that a number of synthetic pyrimidine (Y) and purine (R) nucleotides are recognized by the receptor as substitutes for the natural nucleotides deoxycytidine and deoxyguanosine in a CpG dinucleotide. These studies permitted development of synthetic YpG, CpR and YpR immunostimulatory motifs, and showed divergent nucleotide motif recognition pattern of the receptor. Surprisingly, we found that synthetic immunostimulatory motifs produce different cytokine induction profiles compared with natural CpG motifs. Importantly, we also found that some of these synthetic immunostimulatory motifs show optimal activity in both mouse and human systems without the need to change sequences, suggesting an overriding of the species-dependent specificity of the receptor by the use of synthetic motifs. In the present paper, we review current understanding of structural recognition and functional modulation of TLR9 receptor by second-generation synthetic CpG DNAs and their potential application as wide-spectrum therapeutic agents. PMID- 12773177 TI - Evolutionary relationships, but functional differences, between the Drosophila and human Toll-like receptor families. AB - The Toll receptor was first found to function in the dorsoventral patterning pathway of Drosophila embryos. It is activated by a specific protein ligand, Spatzle, generated at ventral positions in the early embryo. Drosophila Toll (dToll) also functions in innate immune responses to Gram-positive bacteria and fungi, and Spatzle is required for this response. We have shown that Spatzle is necessary and sufficient for activation of the dToll pathway, and that it probably acts by cross-linking two molecules of Toll to form homodimers. In the present paper, we contrast this mode of regulation with that proposed for the vertebrate Toll-like receptor family, which mediate analogous responses to pathogen pattern antigens. In contrast with dToll, these receptors appear to be activated by direct exposure to pathogen patterns, such as peptidoglycan and lipopolysaccharide. We discuss the evolutionary basis of this functional divergence of the vertebrate and invertebrate Toll-like receptors. PMID- 12773178 TI - Toll-like receptor 4 signalling: new perspectives on a complex signal transduction problem. AB - We previously reported that Toll-like receptor-2 (TLR2) agonists induce expression of a more limited repertoire of pro-inflammatory genes than TLR4 agonists. Murine macrophages stimulated with the TLR4 agonist, Escherichia coli lipopolysaccharide, induced signal transducer and activator of transcription 1 ('STAT1') tyrosine phosphorylation that was secondary to the autocrine/paracrine action of interferon (IFN)-beta, an immediate early gene. In contrast, TLR2 agonists failed to activate IFN-beta gene expression. TLR4-induced IFN-beta mRNA was found to be MyD88- and PKR (double-stranded RNA-dependent protein kinase) independent, but TIRAP (Toll/interleukin-1 receptor domain-containing adapter protein)/Mal (MyD88-adapter-like)-dependent. In the present paper, we outline the recent controversy over the role of TIRAP/Mal in TLR2 and TLR4 signalling in the context of the current molecular tools used for such studies. Collectively, our findings provide the first mechanistic basis for differential patterns of gene expression activated by TLR4 and TLR2 agonists. PMID- 12773179 TI - Initiation of archaeal DNA replication. AB - The identification of DNA as the genetic material and the elucidation of its structure by Watson and Crick [Watson and Crick, (1953) Nature (London) 171, 737 738], which has its 50th anniversary this year, first suggested the simple elegance with which the problem of passing on precise genetic information from one generation to the next could be solved. Semi-conservative replication is perhaps one of the simplest biological concepts to explain and understand. However, despite an enormous amount of effort in the intervening years, details of the way in which this process is regulated and performed are still unclear in many organisms. Recent work suggests that, due to their simplicity, the Archaea may make a good model for understanding some of the aspects of eukaryotic replication that still elude us. PMID- 12773180 TI - DNA replication in the hyperthermophilic archaeon Sulfolobus solfataricus. AB - Studies of the DNA-replication machinery of Archaea have revealed striking similarities to that of eukaryotes. Indeed, it appears that in most cases Archaea possess a simplified version of the eukaryotic replication apparatus. Studies of Archaea are therefore shedding light on the fundamental processes of DNA replication in both domains of life. PMID- 12773181 TI - Halophilic adaptation of protein-DNA interactions. AB - Pyrococcus woesei ( Pw ) is an archaeal organism adapted to living in conditions of elevated salt and temperature. Thermodynamic data reveal that the interaction between the TATA-box-binding protein (TBP) from this organism and DNA has an entirely different character to the same interaction in mesophilic counterparts. In the case of the Pw TBP, the affinity of its interaction with DNA increases with increasing salt concentration. The opposite effect is observed in all known mesophilic protein-DNA interactions. The halophilic behaviour can be attributed to sequestration of cations into the protein-DNA complex. By mutating residues in the Pw TBP DNA-binding site, potential sites of cation interaction can be removed. These mutations have a significant effect on the binding characteristics, and the halophilic nature of the Pw TBP-DNA interaction can be reversed, and made to resemble that of a mesophile, in just three mutations. The genes of functionally homologous proteins in organisms existing in different environments show that adaptation is most often accompanied by mutation of an existing protein. However, the importance of any individual residue to a phenotypic characteristic is usually difficult to assess amongst the multitude of changes that occur over evolutionary time. Since the halophilic nature of this protein can be attributed to only three mutations, this reveals that the important phenotype of halophilicity could be rapidly acquired in evolutionary time. PMID- 12773182 TI - The chaperonins: perspectives from the Archaea. AB - Heat-shock protein (Hsp) 60 chaperones are almost ubiquitous and almost always essential. They can be divided on the basis of sequence homology into two broad types: group I (found in bacteria, mitochondria and chloroplasts) and group II (found in Archaea and the eukaryotic cytosol). Of the two, the group I chaperones are the better understood. Data on their structure, mechanism of action and cellular role will be briefly presented. The group II chaperones are less well studied. In eukaryotes they form large complexes with 8-fold symmetry containing eight different subunits, all of which are essential. They appear to have a major role in the folding of actin and tubulin, although they may also act on other substrates. No crystal structures are available for these complexes. The situation in the Archaea is simpler, with organisms containing between one and three genes for these chaperones. A 2.6 A structure exists for one archaeal group II chaperone complex. Some progress has been made in defining the reaction cycle of the archaeal group II chaperones and this has shown that they have some properties distinct from the group I chaperones. To date, the in vivo role and importance of the archaeal group II Hsp60 chaperones has not been determined. We have now shown that in the halophilic archaeon Haloferax volcanii not all the genes for these proteins are essential. Further analysis of these proteins in the Archaea should be very productive in yielding more information about these important chaperones and their cellular functions. PMID- 12773183 TI - The archaeal twin-arginine translocation pathway. AB - The twin-arginine translocation (Tat) pathway is a system with the unique ability to export proteins in a fully folded conformation. Its main components are TatA, TatB and TatC, all of which are required for Tat-dependent export. The Tat pathway is found in several Archaea, and in most of them a moderate number of predicted Tat-dependent substrates are present. Putative substrates include those binding cofactors such as iron-sulphur clusters and molybdopterin. In these Archaea, the role of the Tat pathway seems to be similar to that of bacteria: the export of a small subset of proteins that fold before translocation across the cytoplasmic membrane. The exception to this is the Tat system of the halophilic archaeon Halobacterium sp. NRC-1. In this organism, the majority of extra cytoplasmic proteins are predicted to use the Tat pathway, which is, most likely, a specific adaptation to its particular lifestyle in highly saline conditions. PMID- 12773184 TI - Archaeal DNA repair: paradigms and puzzles. AB - It is now generally accepted that the Archaea share many similarities in their information-processing pathways with eukarya. Archaeal and eukaryal DNA replication and transcriptional machineries show particularly striking similarities, and the archaeal processes have been used extensively as simpler models of the much more complex eukaryal ones. Archaeal DNA-repair pathways are not yet well characterized, and their relationship with repair pathways in bacteria and eukarya are still open to question. There are also strong distinctions between the major subdivisions crenarchaea and euryarchaea within the archaeal domain. This review highlights some of these similarities and differences using specific examples arising from our studies of the double stranded and single-stranded DNA-binding proteins and the repair endonuclease XPF in the crenarchaeote Sulfolobus solfataricus. PMID- 12773185 TI - Repair of UV damage in Halobacterium salinarum. AB - Halobacterium is one of the few known Archaea that tolerates high levels of sunlight in its natural environment. Photoreactivation is probably its most important strategy for surviving UV irradiation and we have shown that both of the major UV photoproducts, cyclobutane pyrimidine dimers (CPDs) and (6-4) photoproducts, can be very efficiently repaired by photoreactivation in this organism. There are two putative photolyase gene homologues in the published genome sequence of Halobacterium sp. NRC-1. We have made a mutant deleted in one of these, phr2, and confirmed that this gene codes for a CPD photolyase. (6-4) photoproducts are still photoreactivated in the mutant so we are currently establishing whether the other homologue, phr1, codes for a (6-4) photolyase. We have also demonstrated an excision repair capacity that operates in the absence of visible light but the nature of this pathway is not yet known. There is probably a bacteria-type excision-repair mechanism, since homologues of uvrA, uvrB, uvrC and uvrD have been identified in the Halobacterium genome. However, there are also homologues of eukaryotic nucleotide-excision-repair genes ( Saccharomyces cerevisiae RAD3, RAD25 and RAD2 ) so there may be multiple repair mechanisms for UV damage in Halobacterium. PMID- 12773186 TI - Uracil recognition by archaeal family B DNA polymerases. AB - Archaeal family-B DNA polymerases possess a novel uracil-sensing mechanism. A specialized pocket scans the template, ahead of the replication fork, for the presence of uracil; on encountering this base, DNA synthesis is stalled. The structural basis for uracil recognition by polymerases is described and compared with other uracil-recognizing enzymes (uridine-triphosphate pyrophophatases and uracil-DNA glycosylases). Remarkably, protein-protein interactions between all three archaeal uracil sensors are observed; possibly the enzymes co-operate to efficiently eliminate uracil from archaeal genomes. PMID- 12773187 TI - Homologous recombination in Archaea: new Holliday junction helicases. AB - Homologous recombination (HR) maintains genome stability by promoting high fidelity DNA repair. Several recent reports have established that the primary function of HR enzymes is to underpin DNA replication, resetting forks that are blocked or collapsed at sites of DNA damage remote from replication origins. These functions are crucial to ensuring that genomes are transmitted successfully into subsequent generations of cells. Enzymes of HR have been unearthed in all three domains of life: bacteria, Archaea and eukarya. Helicases that specifically unwind branched DNA molecules are pivotal in linking HR and DNA replication in bacteria. However, knowledge of helicases with these functions in eukaryotes is vague and is wholly absent in Archaea. We are using the archaeal species Methanothermobacter thermautotrophicus to identify new DNA helicases of homologous recombination. PMID- 12773188 TI - Genetic analysis of homologous recombination in Archaea: Haloferax volcanii as a model organism. AB - Homologous recombination is a fundamental cellular process that rearranges genes both within and between chromosomes, promotes repair of damaged DNA and underpins replication. Much of our understanding of recombination stems from pioneering studies of bacterial and eukaryotic systems such as Escherichia coli and Saccharomyces cerevisiae. Since most archaeal species are extremophilic and difficult to cultivate, current knowledge of recombination in the Archaea is confined largely to comparative genomics and biochemistry. A clear view of what we can learn will not emerge until genetic and molecular systems have been established. We are developing such systems using Haloferax volcanii as a model organism, as it can be cultivated in the laboratory with ease and offers great potential for establishing tractable and informative genetic systems. PMID- 12773189 TI - It can be a complicated life being an enzyme. AB - It is becoming increasingly apparent that many well-known enzymes have alternative non-enzymic functions. Similarly, several proteins that were identified as having non-catalytic functions were subsequently found to have enzyme activities. Some examples are considered to illustrate the diversity of alternative functions. The semicarbazide-sensitive amine oxidase (EC 1.4.3.6) is considered in more depth as an example. It was originally believed to be a detoxifying enzyme, but the reaction products may have important signalling functions. Furthermore, this enzyme, from some sources, also behaves as a vascular-adhesion protein. Finally, the challenges posed by such multiplicity of functions for the interpretation of genetic deletion, in vivo inhibition and the development of functional protein databases are briefly considered. PMID- 12773190 TI - Immunoglobulin-binding domains: Protein L from Peptostreptococcus magnus. AB - Protein L is a multidomain cell-wall protein isolated from Peptostreptococcus magnus. It belongs to a group of proteins that contain repeated domains that are able to bind to Igs without stimulating an immune response, the most characterized of this group being Protein A ( Staphylococcus aureus ) and Protein G ( Streptococcus ). Both of these proteins bind predominantly to the interface of C(H)2-C(H)3 heavy chains, while Protein L binds exclusively to the V(L) domain of the kappa -chain. The function of these proteins in vivo is not clear but it is thought that they enable the bacteria to evade the host's immune system. Two binding sites for kappa -chain on a single Ig-binding domain from Protein L have recently been reported and we give evidence that one site has a 25-55-fold higher affinity for kappa -chain than the second site. PMID- 12773191 TI - Implications of enzyme kinetics. AB - Of the many examples of oscillatory kinetic behaviour known, several are briefly reviewed, including those of glycolysis, the peroxidase-oxidase reaction and oscillations in cellular calcium concentration. It is shown that simple mathematical models employing allosteric rate laws are sufficient to explain the instability of the steady state and the appearance of sustained oscillations. The cAMP-signalling systems of cellular slime moulds and the dynamics of intracellular calcium oscillations illustrate the importance of such oscillophores to inter- and intra-cellular communication and differentiation. PMID- 12773192 TI - Exploring the structure and function of zinc metallopeptidases: old enzymes and new discoveries. AB - Neprilysin [or neutral endopeptidase (NEP)] and angiotensin-converting enzyme (ACE) are zinc metallopeptidases involved in the extracellular metabolism of biologically active peptides. Recent genomic advances have led to the identification of novel homologues of each of these ectoenzymes and new physiological and pathological roles are emerging for them. The structures of each of these peptidases have recently been solved providing insight into their distinct catalytic sites. In addition to its originally identified role in neuropeptide metabolism in the nervous system, NEP is implicated in regulation of the cardiovascular system and is protective in prostate and certain other cancers. Hence the cellular concentration of NEP is critical to tissue homoeostasis. Most recently, NEP has been shown to exert neuroprotective actions, principally through its ability to catabolize the neurotoxic Alzheimer's amyloid peptide. The only known homologue of ACE, termed ACE2, is critical to cardiovascular function, but its physiological substrates and precise metabolic roles remain to be elucidated. Other members of these growing metallopeptidase families await further characterization and possible exploitation as therapeutic targets. PMID- 12773193 TI - How to tell the time: the regulation of phosphoenolpyruvate carboxylase in Crassulacean acid metabolism (CAM) plants. AB - Crassulacean acid metabolism (CAM) plants exhibit persistent circadian rhythms of CO(2) metabolism. These rhythms are driven by changes in the flux through phosphoenolpyruvate carboxylase, which is regulated by reversible phosphorylation in response to a circadian oscillator. This article reviews progress in our understanding of the circadian expression of phosphoenolpyruvate carboxylase kinase. PMID- 12773194 TI - Human porphobilinogen deaminase mutations in the investigation of the mechanism of dipyrromethane cofactor assembly and tetrapyrrole formation. AB - Porphobilinogen deaminase mutants that cause acute intermittent porphyria have been investigated as recombinant proteins expressed in Escherichia coli, yielding important insight into the mechanism of dipyrromethane cofactor assembly and tetrapyrrole chain polymerization. A mutation that affects a key catalytic residue, D99G, results in an inactive holo -protein that exists as a complex with two substrate molecules covalently bound to the dipyrromethane cofactor arising from the reaction between the apo -protein and pre-uroporphyrinogen. The R149Q mutant is also devoid of catalytic activity but the mutant protein is unable to assemble the dipyrromethane cofactor from pre-uroporphyrinogen and persists as an unstable, heat-labile apo -protein. The mutant, R173Q, has very low activity and, like R149Q, also exhibits largely as an apo -protein. The inability to reconstitute either R149Q or R173Q with exogenous pre-uroporphyrinogen confirms the importance of these two arginine residues for dipyrromethane cofactor assembly. In contrast, the mutant R167Q exists as a holo -enzyme but the catalytic cycle is severely compromised, leading to the accumulation of stable enzyme-substrate intermediates from the catalytic cycle. PMID- 12773195 TI - Clathrin-coated vesicle formation: a paradigm for coated-vesicle formation. AB - Clathrin-coated pits are the major ports of entry into the cell and are responsible for the internalization of a variety of biologically important macromolecules. These transport intermediates form as a result of the co ordinated assembly of a number of cytosolic proteins on to the membrane which results in specific cargo recruitment. We have used a variety of approaches including permeabilized cell assays and light and electron microscopy to identify and characterize the proteins and enzymes involved in coated vesicle formation. PMID- 12773196 TI - Cardiovascular and hormonal effects of subcutaneous administration of ghrelin, a novel growth hormone-releasing peptide, in healthy humans. AB - Ghrelin is a novel GH (growth hormone)-releasing peptide isolated from the stomach. The cardiovascular and hormonal effects of the subcutaneous administration of ghrelin in humans remain unknown. Six healthy volunteers each received subcutaneous administration of three doses of ghrelin (1, 5 or 10 microg/kg) and placebo; the order of administration was randomized, and separate doses were given at least 24 h apart. The serum GH level dose-dependently increased from 0.5 +/- 0.4 to 3.6 +/- 2.1 ng/ml (1 microg/kg ghrelin; P=0.99 compared with baseline), 27.1 +/- 12.0 ng/ml (5 microg/kg; P<0.01 compared with baseline) and 45.4 +/- 12.8 ng/ml (10 microg/kg; P<0.01 compared with baseline) 30 min after ghrelin administration. Subcutaneous administration of ghrelin did not significantly alter circulating levels of corticotropin, cortisol, insulin like growth factor-1, noradrenaline or adrenaline, although 10 microg/kg ghrelin slightly increased the prolactin level. No significant changes in heart rate or mean arterial pressure were observed. In contrast, the left ventricular ejection fraction, as assessed by echocardiography, increased dose-dependently from 63.5 +/- 0.6% to 65.1 +/- 0.9% (1 microg/kg ghrelin; P=0.97 compared with baseline), 69.6 +/- 1.3% (5 microg/kg; P<0.01 compared with baseline) and 71.5 +/- 0.9% (10 microg/kg; P<0.01 compared with baseline) 30 min after ghrelin administration. These haemodynamic and hormonal changes were still apparent 60 min after ghrelin injection. In conclusion, subcutaneous administration of ghrelin dose-dependently induced relatively specific GH release and enhanced cardiac performance in humans. PMID- 12773197 TI - A physiological role for alpha2-HS glycoprotein: stimulation of macrophage uptake of apoptotic cells. AB - This comment describes the study by Jersmann and co-workers in this issue of Clinical Science reporting the results of a study of the role of the serum glycoprotein fetuin in the uptake of apoptotic cells by macrophages. They show that fetuin is able to stimulate the macropinocytosis of apoptotic cells in vivo, which would be therapeutically useful following chemotherapy when the increased numbers of apoptotic cells could exceed the capacity of the macrophage network. PMID- 12773198 TI - Central Mucoepidermoid carcinoma of mandible - A case report and review of the literature. AB - BACKGROUND: Primary central mucoepidermoid carcinoma of jaws is a rare lesion comprising 2-3% of all mucoepidermoid carcinomas reported in literature. CASE PRESENTATION: The case presented here illustrates the hypothesis that its specific pathogenesis is unknown. CONCLUSIONS: Mucoepidermoid carcinoma of the jaw is a rare tumour of unknown aetiology. Although about a 100 cases has been reported in literature, the speculation on its aetiopathogenesis has mainly centred on the pluripotential capabilities of the epithelial lining of odontogenic cysts. PMID- 12773199 TI - Development, validity and responsiveness of the Clinical COPD Questionnaire. AB - BACKGROUND: The new Global Obstructive Lung Disease (GOLD) guidelines advice to focus treatment in Chronic Obstructive Pulmonary Disease (COPD) on improvement of functional state, prevention of disease progression and minimization of symptoms. So far no validated questionnaires are available to measure symptom and functional state in daily clinical practice. The aim of this study was to develop and validate the Clinical COPD Questionnaire (CCQ). METHODS: Qualitative research with patients and clinicians was performed to generate possible items to evaluate clinical COPD control. Thereafter, an item reduction questionnaire was sent to 77 international experts. Sixty-seven experts responded and the 10 most important items, divided into 3 domains (symptoms, functional and mental state) were included in the CCQ (scale: 0 = best, 6 = worst). RESULTS: Cross-sectional data were collected from 119 subjects (57 COPD, GOLD stage I-III; 18 GOLD stage 0 and 44 (ex)smokers). Cronbach's alpha was high (0.91). The CCQ scores in patients (GOLD 0-III) were significantly higher than in healthy (ex)smokers. Furthermore, significant correlations were found between the CCQ total score and domains of the SF-36 (rho = 0.48 to rho = 0.69) and the SGRQ (rho = 0.67 to rho = 0.72). In patients with COPD, the correlation between the CCQ and FEV1%pred was rho =-0.49. Test-retest reliability was determined in 20 subjects in a 2-week interval (Intra Class Coefficient = 0.94). Thirty-six smokers with and without COPD showed significant improvement in the CCQ after 2 months smoking cessation, indicating the responsiveness of the CCQ. CONCLUSION: The CCQ is a self-administered questionnaire specially developed to measure clinical control in patients with COPD. Data support the validity, reliability and responsiveness of this short and easy to administer questionnaire. PMID- 12773200 TI - Self-reported health-related quality of life in persons with HIV infection: results from a multi-site interview project. AB - BACKGROUND: To examine demographic and behavioral associations with self-reported health-related quality of life (HRQOL) among persons with HIV infection or AIDS. METHODS: Analysis of interviews with persons > or = 18 years of age reported through routine disease surveillance with HIV infection or AIDS to nine state and local health departments from January 1995 through December 1996. Scales were constructed from validated measures of HRQOL, and mean scores were calculated (lower scores signified poorer HRQOL). Measures of HRQOL included Overall Health, Pain, Physical Functioning, Role Functioning, Social Functioning, Mental Health, Energy/Fatigue, and Cognitive Functioning. Differences in HRQOL were examined by various demographic and behavioral factors, including taking antiretroviral medication. RESULTS: HRQOL data were available for 3778 persons. Factors associated with lower HRQOL scores included older age, female sex, black or Hispanic race/ethnicity, injection drug use, lower education and income, no private health insurance, and lower CD4 count. In multivariate analysis, lower CD4 count was the factor most consistently associated with lower HRQOL. Taking antiretroviral medication was not associated with differences in HRQOL regardless of CD4 count. CONCLUSIONS: Perception of HRQOL varied in a population with HIV infection or AIDS. On most HRQOL measures, lower CD4 count was associated with lower HRQOL. Measurement of HRQOL can assist in understanding the long-term effects of disease and treatment on persons with HIV. PMID- 12773201 TI - Phase angle correlates with n-3 fatty acids and cholesterol in red cells of Nigerian children with sickle cell disease. AB - OBJECTIVE: To determine the cholesterol content and fatty acid composition of red cell membrane phospholipids (PL) of children with sickle cell disease (SCD) and to correlate these levels with whole body phase angle that is related to the integrity and function of cell membranes. STUDY DESIGN: Blood samples were obtained from 69 children with SCD and 72 healthy age- and gender-matched controls in Nigeria for the determination of the cholesterol content and proportions of fatty acids in red cell PL. Bioelectrical impedance analysis was used to obtain resistance (R) and reactance (Xc) from which phase angle was calculated as arctan Xc/R. Cholesterol (normalized to lipid phosphorus) and the proportions of individual fatty acids were correlated with phase angle. RESULTS: The proportions of palmitic (p < 0.001), stearic acid (p = 0.003) and cholesterol (p < 0.001) were significantly higher in the red cells of children with SCD, whereas the proportions of arachidonic acid and docosahexaenoic acid were reduced (p = 0.03 and < 0.001, respectively) compared to controls. The phase angle was inversely correlated with the proportions of palmitic acid (p = 0.03) and oleic acid (p < 0.001) and cholesterol (p = 0.003). Three n-3 polyunsaturated fatty acids-eicosapentaenoic acid, docosapentaenoic acid and docosahexaenoic acid- were positively correlated with phase angle (p < 0.001). CONCLUSIONS: The fatty acid composition and cholesterol content of tissue membranes in SCD correlate with the phase shift measured by bioelectrical impedance analysis. Phase angle measurements may provide a non-invasive method for monitoring interventions aimed at altering the lipid composition of membranes. PMID- 12773203 TI - A step too far? Making health equity interventions in Namibia more sufficient. AB - BACKGROUND: Equality of health status is the health equity goal being pursued in developed countries and advocated by development agencies such as WHO and The Rockefeller Foundation for developing countries also. Other concepts of fair distribution of health such as equity of access to medical care may not be sufficient to equalise health outcomes but, nevertheless, they may be more practical and effective in advancing health equity in developing countries. METHODS: A framework for relating health equity goals to development strategies allowing progressive redistribution of primary health care resources towards the more deprived communities is formulated. The framework is applied to the development of primary health care in post-independence Namibia. RESULTS: In Namibia health equity has been advanced through the progressive application of health equity goals of equal distribution of primary care resources per head, equality of access for equal met need and equality of utilisation for equal need. For practical and efficiency reasons it is unlikely that health equity would have been advanced further or more effectively by attempting to implement the goal of equality of health status. CONCLUSION: The goal of equality of health status may not be appropriate in many developing country situations. A stepwise approach based on progressive redistribution of medical services and resources may be more appropriate. This conclusion challenges the views of health economists who emphasise the need to select a single health equality goal and of development agencies which stress that equality of health status is the most important dimension of health equity. PMID- 12773202 TI - Frequent promoter methylation of CDH1, DAPK, RARB, and HIC1 genes in carcinoma of cervix uteri: its relationship to clinical outcome. AB - BACKGROUND: Cervical cancer (CC), a leading cause of cancer-related deaths in women worldwide, has been causally linked to genital human papillomavirus (HPV) infection. Although a host of genetic alterations have been identified, molecular basis of CC development is still poorly understood. RESULTS: We examined the role of promoter hypermethylation, an epigenetic alteration that is associated with the silencing tumor suppressor genes in human cancer, by studying 16 gene promoters in 90 CC cases. We found a high frequency of promoter methylation in CDH1, DAPK, RARB, and HIC1 genes. Correlation of promoter methylation with clinical characteristics and other genetic changes revealed the following: a) overall promoter methylation was higher in more advanced stage of the disease, b) promoter methylation of RARB and BRCA1 predicted worse prognosis, and c) the HIC1 promoter methylation was frequently seen in association with microsatellite instability. Promoter methylation was associated with gene silencing in CC cell lines. Treatment with methylation or histone deacetylation-inhibiting agents resulted in profound reactivation of gene expression. CONCLUSIONS: These results may have implications in understanding the underlying epigenetic mechanisms in CC development, provide prognostic indicators, and identify important gene targets for treatment. PMID- 12773204 TI - Electromagnetic interference of GSM mobile phones with the implantable deep brain stimulator, ITREL-III. AB - BACKGROUND: The purpose was to investigate mobile phone interference with implantable deep brain stimulators by means of 10 different 900 Mega Hertz (MHz) and 10 different 1800 MHz GSM (Global System for Mobile Communications) mobile phones. METHODS: All tests were performed in vitro using a phantom especially developed for testing with deep brain stimulators. The phantom was filled with liquid phantom materials simulating brain and muscle tissue. All examinations were carried out inside an anechoic chamber on two implants of the same type of deep brain stimulator: ITREL-III from Medtronic Inc., USA. RESULTS: Despite a maximum transmitted peak power of mobile phones of 1 Watt (W) at 1800 MHz and 2 W at 900 MHz respectively, no influence on the ITREL-III was found. Neither the shape of the pulse form changed nor did single pulses fail. Tests with increased transmitted power using CW signals and broadband dipoles have shown that inhibition of the ITREL-III occurs at frequency dependent power levels which are below the emissions of GSM mobile phones. The ITREL-III is essentially more sensitive at 1800 MHz than at 900 MHz. Particularly the frequency range around 1500 MHz shows a very low interference threshold. CONCLUSION: These investigations do not indicate a direct risk for ITREL-III patients using the tested GSM phones. Based on the interference levels found with CW signals, which are below the mobile phone emissions, we recommend similar precautions as for patients with cardiac pacemakers: 1. The phone should be used at the ear at the opposite side of the implant and 2. The patient should avoid carrying the phone close to the implant. PMID- 12773206 TI - Consumption of fruits and vegetables in relation to the risk of developing acute coronary syndromes; the CARDIO2000 case-control study. AB - BACKGROUND: The relation between diet and human health has long been investigated. The aim of this work is to evaluate the association between CHD risk and the consumption of fruit and vegetable, in a large sample of cardiac patients and controls. METHODS: Stratified sampling from all Greek regions, consisted of 848 (700 males, 58 +/- 10 years old and 148 females, 65 +/- 9 years old) randomly selected patients, admitted to the cardiology clinic for a first event of an acute coronary syndrome (ACS). In addition we selected 1078 frequency paired, by sex-age-region, controls in the same hospitals but without any clinical suspicion of CHD. Using validated food-frequency questionnaires we assessed total diet, including fruit and vegetable intake, on a weekly basis. Multiple logistic regression analysis estimated the relative risk of developing ACS by level of fruits and vegetables intake after taking into account the effect of several potential confounders. RESULTS: Data analysis revealed that the benefit of fruit or vegetable consumption increases proportionally by the number of servings consumed (P for trend < 0.001). After adjusting for the conventional cardiovascular risk factors, those in the upper quintile of fruit consumption (5 or more items/day) had 72% lower risk for CHD (odds ratio = 0.28, 95% CI 0.11 0.54, P < 0.001), compared with those in the lowest quintile of intake (< 1 items/day). Similarly, consumption of vegetable more than 3 days/week was associated with 70% lower risk for CHD (odds ratio = 0.30, 95% CI 0.22-0.40, P < 0.001), compared with those that they did not consume vegetables. Of particular interest, a 10% reduction in coronary risk was observed for every one piece of fruit consumed per day (odds ratio = 0.90, 95% CI 0.85-0.97, P = 0.004). CONCLUSIONS: Consumption of fruits and vegetables seems to offer significant protection against CHD. PMID- 12773205 TI - Alpha-tocopherol modulates the low density lipoprotein receptor of human HepG2 cells. AB - The aim of this study was to determine the effects of vitamin E (alpha tocopherol) on the low density lipoprotein (LDL) receptor, a cell surface protein which plays an important role in controlling blood cholesterol. Human HepG2 hepatoma cells were incubated for 24 hours with increasing amounts of alpha, delta, or gamma-tocopherol. The LDL receptor binding activity, protein and mRNA, 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase mRNA, cell cholesterol and cell lathosterol were measured. The effect of alpha-tocopherol was biphasic. Up to a concentration of 50 microM, alpha-tocopherol progressively increased LDL receptor binding activity, protein and mRNA to maximum levels 2, 4 and 6-fold higher than control, respectively. The HMG-CoA reductase mRNA and the cell lathosterol concentration, indices of cholesterol synthesis, were also increased by 40% over control by treatment with 50 microM alpha-tocopherol. The cell cholesterol concentration was decreased by 20% compared to control at 50 microM alpha-tocopherol. However, at alpha-tocopherol concentrations higher than 50 microM, the LDL receptor binding activity, protein and mRNA, the HMG-CoA reductase mRNA and the cell lathosterol and cholesterol concentrations all returned to control levels. The biphasic effect on the LDL receptor was specific for alpha-tocopherol in that delta and gamma-tocopherol suppressed LDL receptor binding activity, protein and mRNA at all concentrations tested despite the cells incorporating similar amounts of the three homologues. In conclusion, alpha tocopherol, exhibits a specific, concentration-dependent and biphasic "up then down" effect on the LDL receptor of HepG2 cells which appears to be at the level of gene transcription. Cholesterol synthesis appears to be similarly affected and the cell cholesterol concentration may mediate these effects. PMID- 12773207 TI - Polymorphism in two merozoite surface proteins of Plasmodium falciparum isolates from Gabon. AB - BACKGROUND: Plasmodium falciparum antigenic diversity and polymorphism confuses the issue of antimalarial vaccine development. Merozoite surface protein (MSP)-1 and -2 are two highly polymorphic vaccine candidates. Characterisation of their precise polymorphism in endemic regions may facilitate the design of an effective vaccine. METHODS: Isolates obtained in 52 Gabonese children presenting with uncomplicated malaria were genotyped by nested-PCR of msp-1 block 2, and msp-2 block 3, to analyze both parasite population polymorphism and clone fluctuations. RESULTS: Twenty-five and 19 different alleles were respectively obtained for msp 1 and msp-2 loci, the RO33 family of msp-1 being poorly polymorphic. Four cases of non-random distribution of alleles were reported of the FC27, and/or 3D7 families of msp-2. All but two isolates were composed of more than one genotype, and the multiplicity of infection (MOI) was 4.0. Neither parasite density nor age was related to MOI. Clone fluctuations were studied for ten subjects who were sampled again at reappearance of parasites in blood. Disappearance and reappearance of alleles were observed following treatment, suggesting difficulties in assessing polymorphism and in distinguishing reinfection from recrudescence. CONCLUSION: P. falciparum polymorphism is extensive in Southeast Gabon, and most of infections are composed of multiple clones. The fluctuation of clones contributes to parasite diversity. PMID- 12773208 TI - Prevention of poxvirus infection by tetrapyrroles. AB - BACKGROUND: Prevention of poxvirus infection is a topic of great current interest. We report inhibition of vaccinia virus in cell culture by porphyrins and phthalocyanines. Most previous work on the inhibition of viruses with tetrapyrroles has involved photodynamic mechanisms. The current study, however, investigates light-independent inhibition activity. METHODS: The Western Reserve (WR) and International Health Department-J (IHD-J) strains of vaccinia virus were used. Virucidal and antiviral activities as well as the cytotoxicity of test compounds were determined. RESULTS: Examples of active compounds include zinc protoporphyrin, copper hematoporphyrin, meso(2,6-dihydroxyphenyl)porphyrin, the sulfonated tetra-1-naphthyl and tetra-1-anthracenylporphyrins, selected sulfonated derivatives of halogenated tetraphenyl porphyrins and the copper chelate of tetrasulfonated phthalocyanine. EC50 values for the most active compounds are as low as 0.05 microg/mL (40 nM). One of the most active compounds was the neutral meso(2,6-dihydroxyphenyl)porphyrin, indicating that the compounds do not have to be negatively charged to be active. CONCLUSIONS: Porphyrins and phthalocyanines have been found to be potent inhibitors of infection by vaccinia virus in cell culture. These tetrapyrroles were found to be active against two different virus strains, and against both enveloped and non-enveloped forms of the virus, indicating that these compounds may be broadly effective in their ability to inhibit poxvirus infection. PMID- 12773210 TI - Population health metrics: crucial inputs to the development of evidence for health policy. AB - Valid, reliable and comparable measures of the health states of individuals and of the health status of populations are critical components of the evidence base for health policy. We need to develop population health measurement strategies that coherently address the relationships between epidemiological measures (such as risk exposures, incidence, and mortality rates) and multi-domain measures of population health status, while ensuring validity and cross-population comparability.Studies reporting on descriptive epidemiology of major diseases, injuries and risk factors, and on the measurement of health at the population level - either for monitoring trends in health levels or inequalities or for measuring broad outcomes of health systems and social interventions - are not well-represented in traditional epidemiology journals, which tend to concentrate on causal studies and on quasi-experimental design. In particular, key methodological issues relating to the clear conceptualisation of, and the validity and comparability of measures of population health are currently not addressed coherently by any discipline, and cross-disciplinary debate is fragmented and often conducted in mutually incomprehensible language or paradigms. Population health measurement potentially bridges a range of currently disjoint fields of inquiry relating to health: biology, demography, epidemiology, health economics, and broader social science disciplines relevant to assessment of health determinants, health state valuations and health inequalities.This new journal will focus on the importance of a population based approach to measurement as a way to characterize the complexity of people's health, the diseases and risks that affect it, its distribution, and its valuation, and will attempt to provide a forum for innovative work and debate that bridge the many fields of inquiry relevant to population health in order to contribute to the development of valid and comparable methods for the measurement of population health and its determinants. PMID- 12773209 TI - Survival of the fittest before the beginning of life: selection of the first oligonucleotide-like polymers by UV light. AB - BACKGROUND: A key event in the origin of life on this planet has been formation of self-replicating RNA-type molecules, which were complex enough to undergo a Darwinian-type evolution (origin of the "RNA world"). However, so far there has been no explanation of how the first RNA-like biopolymers could originate and survive on the primordial Earth. RESULTS: As condensation of sugar phosphates and nitrogenous bases is thermodynamically unfavorable, these compounds, if ever formed, should have undergone rapid hydrolysis. Thus, formation of oligonucleotide-like structures could have happened only if and when these structures had some selective advantage over simpler compounds. It is well known that nitrogenous bases are powerful quenchers of UV quanta and effectively protect the pentose-phosphate backbones of RNA and DNA from UV cleavage. To check if such a protection could play a role in abiogenic evolution on the primordial Earth (in the absence of the UV-protecting ozone layer), we simulated, by using Monte Carlo approach, the formation of the first oligonucleotides under continuous UV illumination. The simulations confirmed that UV irradiation could have worked as a selective factor leading to a relative enrichment of the system in longer sugar-phosphate polymers carrying nitrogenous bases as UV-protectors. Partial funneling of the UV energy into the condensation reactions could provide a further boost for the oligomerization. CONCLUSION: These results suggest that accumulation of the first polynucleotides could be explained by their abiogenic selection as the most UV-resistant biopolymers. PMID- 12773211 TI - Estimating the prevalence of breast cancer using a disease model: data problems and trends. AB - BACKGROUND: Health policy and planning depend on quantitative data of disease epidemiology. However, empirical data are often incomplete or are of questionable validity. Disease models describing the relationship between incidence, prevalence and mortality are used to detect data problems or supplement missing data. Because time trends in the data affect their outcome, we compared the extent to which trends and known data problems affected model outcome for breast cancer. METHODS: We calculated breast cancer prevalence from Dutch incidence and mortality data (the Netherlands Cancer Registry and Statistics Netherlands) and compared this to regionally available prevalence data (Eindhoven Cancer Registry, IKZ). Subsequently, we recalculated the model adjusting for 1) limitations of the prevalence data, 2) a trend in incidence, 3) secondary primaries, and 4) excess mortality due to non-breast cancer deaths. RESULTS: There was a large discrepancy between calculated and IKZ prevalence, which could be explained for 60% by the limitations of the prevalence data plus the trend in incidence. Secondary primaries and excess mortality had relatively small effects only (explaining 17% and 6%, respectively), leaving a smaller part of the difference unexplained. CONCLUSION: IPM models can be useful both for checking data inconsistencies and for supplementing incomplete data, but their results should be interpreted with caution. Unknown data problems and trends may affect the outcome and in the absence of additional data, expert opinion is the only available judge. PMID- 12773212 TI - A generic model for the assessment of disease epidemiology: the computational basis of DisMod II. AB - Epidemiology as an empirical science has developed sophisticated methods to measure the causes and patterns of disease in populations. Nevertheless, for many diseases in many countries only partial data are available. When the partial data are insufficient, but data collection is not an option, it is possible to supplement the data by exploiting the causal relations between the various variables that describe a disease process. We present a simple generic disease model with incidence, one prevalent state, and case fatality and remission. We derive a set of equations that describes this disease process and allows calculation of the complete epidemiology of a disease given a minimum of three input variables. We give the example of asthma with age-specific prevalence, remission, and mortality as inputs. Outputs are incidence and case fatality, among others. The set of equations is embedded in a software package called 'DisMod II', which is made available to the public domain by the World Health Organization. PMID- 12773213 TI - A longitudinal analysis of the risk factors for diabetes and coronary heart disease in the Framingham Offspring Study. AB - BACKGROUND: The recent trends in sedentary life-styles and weight gain are likely to contribute to chronic conditions such as hypertension, diabetes, and cardiovascular diseases. The temporal sequence and pathways underlying these conditions can be modeled using the knowledge from the biomedical and social sciences. METHODS: The Framingham Offspring Study in the U.S. collected information on 5124 subjects at baseline, and 8, 12, 16, and 20 years after the baseline. Dynamic random effects models were estimated for the subjects' weight, LDL and HDL cholesterol, and blood pressure using 4 time observations. Logistic and probit models were estimated for the probability of diabetes and coronary heart disease (CHD) events. RESULTS: The subjects' age, physical activity, alcohol consumption, and cigarettes smoked were important predictors of the risk factors. Moreover, weight and height were found to differentially affect the probabilities of diabetes and CHD events; body weight was positively associated with the risk of diabetes while taller individuals had lower risk of CHD events. CONCLUSION: The results showed the importance of joint modeling of body weight, LDL and HDL cholesterol, and blood pressure that are risk factors for diabetes and CHD events. Lower body weight and LDL concentrations and higher HDL levels achieved via physical exercise are likely to reduce diabetes and CHD events. PMID- 12773214 TI - Monitoring health inequalities: life expectancy and small area deprivation in New Zealand. AB - BACKGROUND: Socioeconomic and ethnic inequalities in health are of great concern, and life expectancy provides a readily understood means of monitoring such inequalities. The objectives of this study are to (1) measure life expectancy by socioeconomic deprivation and ethnicity, and (2) describe trends in the deprivation gradient in life expectancy since the mid-1990s. METHODS: Three years of national mortality data have been combined with mid-point population denominators to produce life tables within nationally determined levels of small area deprivation (NZDep96) for three ethnic group: European, Maori and Pacific peoples. This process has been repeated for the periods 1995-97, 1996-98, 1997-99 and 1998-2000. RESULTS: There was a strong relationship between increasing small area deprivation and decreasing life expectancy. Through the mid- to late 1990s, males living in the most deprived small areas in New Zealand experienced life expectancies at birth approximately nine years less than their counterparts living in the least deprived areas; for females the corresponding difference was under seven years.Maori and Pacific life expectancies at birth were lower than those of Europeans at each level of deprivation.Over the study period (1995-2000) the gradient in life expectancy across deprivation deciles remained stable. CONCLUSION: Small area deprivation analyses of life expectancy could be repeated routinely at regular intervals, which would provide a useful approach to monitoring trends in socioeconomic, geographic, ethnic and gender inequalities in mortality. PMID- 12773215 TI - PopMod: a longitudinal population model with two interacting disease states. AB - This article provides a description of the population model PopMod, which is designed to simulate the health and mortality experience of an arbitrary population subjected to two interacting disease conditions as well as all other "background" causes of death and disability. Among population models with a longitudinal dimension, PopMod is unique in modelling two interacting disease conditions; among the life-table family of population models, PopMod is unique in not assuming statistical independence of the diseases of interest, as well as in modelling age and time independently. Like other multi-state models, however, PopMod takes account of "competing risk" among diseases and causes of death.PopMod represents a new level of complexity among both generic population models and the family of multi-state life tables. While one of its intended uses is to describe the time evolution of population health for standard demographic purposes (e.g. estimates of healthy life expectancy), another prominent aim is to provide a standard measure of effectiveness for intervention and cost effectiveness analysis. PopMod, and a set of related standard approaches to disease modelling and cost-effectiveness analysis, will facilitate disease modelling and cost-effectiveness analysis in diverse settings and help make results more comparable. PMID- 12773216 TI - Why prioritize when there isn't enough money? AB - In an informal address to the 4th International Conference on Priorities in Health (Oslo, 23 September 2002), Professor Jeffrey Sachs - Chairperson of the WHO Commission on Macroeconomics and Health - maintained that the real causes of the inability of the world's poorest people to receive help for the lethal diseases that burden them did not include the "usual suspects" (corruption, mismanagement, and wrong priorities). Rather, the root cause was argued to be an inherent lack of money, indicating that the burden of disease would be lifted only if rich countries gave more money to poor ones.Without taking exception to anything that Sachs said in his address, there nevertheless remain a number of justifications for efforts to improve priority setting in the face of severely shortages of resources, including the following three defenses: prioritization is needed if we are to know that prioritization is insufficient; prioritization is most important when there is little money; prioritization can itself increase resources. PMID- 12773217 TI - Separate spheres and indirect benefits. AB - On any plausible account of the basis for health care resource prioritization, the benefits and costs of different alternative resource uses are relevant considerations in the prioritization process. Consequentialists hold that the maximization of benefits with available resources is the only relevant consideration. Non-consequentialists do not reject the relevance of consequences of benefits and costs, but insist that other considerations, and in particular the distribution of benefits and costs, are morally important as well. Whatever one's particular account of morally justified standards for the prioritization of different health interventions, we must be able to measure those interventions' benefits and costs.There are many theoretical and practical difficulties in that measurement, such as how to weigh extending life against improving health and quality of life as well as how different quality of life improvements should be valued, but they are not my concern here. This paper addresses two related issues in assessing benefits and costs for health resource prioritization. First, should benefits be restricted only to health benefits, or include as well other non health benefits such as economic benefits to employers from reducing the lost work time due to illness of their employees? I shall call this the Separate Spheres problem. Second, should only the direct benefits, such as extending life or reducing disability, and direct costs, such as costs of medical personnel and supplies, of health interventions be counted, or should other indirect benefits and costs be counted as well? I shall call this the Indirect Benefits problem. These two issues can have great importance for a ranking of different health interventions by either a cost/benefit or cost effectiveness analysis (CEA) standard. PMID- 12773218 TI - Econometric estimation of country-specific hospital costs. AB - Information on the unit cost of inpatient and outpatient care is an essential element for costing, budgeting and economic-evaluation exercises. Many countries lack reliable estimates, however. WHO has recently undertaken an extensive effort to collect and collate data on the unit cost of hospitals and health centres from as many countries as possible; so far, data have been assembled from 49 countries, for various years during the period 1973-2000. The database covers a total of 2173 country-years of observations. Large gaps remain, however, particularly for developing countries. Although the long-term solution is that all countries perform their own costing studies, the question arises whether it is possible to predict unit costs for different countries in a standardized way for short-term use. The purpose of the work described in this paper, a modelling exercise, was to use the data collected across countries to predict unit costs in countries for which data are not yet available, with the appropriate uncertainty intervals.The model presented here forms part of a series of models used to estimate unit costs for the WHO-CHOICE project. The methods and the results of the model, however, may be used to predict a number of different types of country specific unit costs, depending on the purpose of the exercise. They may be used, for instance, to estimate the costs per bed-day at different capacity levels; the "hotel" component of cost per bed-day; or unit costs net of particular components such as drugs.In addition to reporting estimates for selected countries, the paper shows that unit costs of hospitals vary within countries, sometimes by an order of magnitude. Basing cost-effectiveness studies or budgeting exercises on the results of a study of a single facility, or even a small group of facilities, is likely to be misleading. PMID- 12773220 TI - Programme costs in the economic evaluation of health interventions. AB - Estimating the costs of health interventions is important to policy-makers for a number of reasons including the fact that the results can be used as a component in the assessment and improvement of their health system performance. Costs can, for example, be used to assess if scarce resources are being used efficiently or whether there is scope to reallocate them in a way that would lead to improvements in population health. As part of its WHO-CHOICE project, WHO has been developing a database on the overall costs of health interventions in different parts of the world as an input to discussions about priority setting.Programme costs, defined as costs incurred at the administrative levels outside the point of delivery of health care to beneficiaries, may comprise an important component of total costs. Cost-effectiveness analysis has sometimes omitted them if the main focus has been on personal curative interventions or on the costs of making small changes within the existing administrative set-up. However, this is not appropriate for non-personal interventions where programme costs are likely to comprise a substantial proportion of total costs, or for sectoral analysis where questions of how best to reallocate all existing health resources, including administrative resources, are being considered.This paper presents a first effort to systematically estimate programme costs for many health interventions in different regions of the world. The approach includes the quantification of resource inputs, choice of resource prices, and accounts for different levels of population coverage. By using an ingredients approach, and making tools available on the World Wide Web, analysts can adapt the programme costs reported here to their local settings. We report results for a selected number of health interventions and show that programme costs vary considerably across interventions and across regions, and that they can contribute substantially to the overall costs of interventions. PMID- 12773219 TI - Costs of stroke and stroke services: Determinants of patient costs and a comparison of costs of regular care and care organised in stroke services. AB - BACKGROUND: Stroke is a major cause of death and long-term disability in Western societies and constitutes a major claim on health care budgets. Organising stroke care in a stroke service has recently been demonstrated to result in better health effects for patients. This paper discusses patient costs after stroke and compares costs between regular and stroke service care. METHODS: Costs were calculated within the framework of the evaluation of three experiments with stroke services in the Netherlands. Cost calculations are base on medical consumption data and actual costs. RESULTS: 598 patients were consecutively admitted to hospital after stroke. The average total costs of care per patient for the 6 month follow-up are estimated at 16,000. Costs are dominated by institutional and accommodation costs. Patients who die after stroke incur less costs. For patients that survive the acute phase, the most important determinants of costs are disability status and having a partner - as they influence patients' stroke careers. These determinants also interact. The most efficient stroke service experiment was most successful in co-ordinating patient flow from hospital to (nursing) home, through capacity planning and efficient discharge procedures. In this region the costs of stroke service care are the same as for regular stroke care. The other experiments suffered from waiting lists for nursing homes and home care, leading to "blocked beds" in hospitals and nursing homes and higher costs of care. Costs of co-ordination are estimated at about 3% of total costs of care. CONCLUSION: This paper demonstrates that by organising care for stroke patients in a stroke service, better health effects can be achieved with the same budget. In addition, it provides insight in need, predisposing and enabling factors that determine costs of care after stroke. PMID- 12773221 TI - Unusual presentation of choriocarcinoma. AB - BACKGROUND: Choriocarcinoma is an aggressive neoplasm arising in the body of the uterus. The disease normally spreads to lung and brain. CASE REPORT: A case of malignant trophoblastic disease with brain metastasis, raised intra cranial pressure and small bowel metastasis presenting with acute abdomen is reported. CONCLUSIONS: Malignant transformation in a hydatidiform mole is rare event. Involvement of gastrointestinal tract is rarer even in presence of disseminated disease. Surgery is the treatment of choice for gastrointestinal complications. PMID- 12773222 TI - Tuberculosis and metastatic carcinoma coexistence in axillary lymph node: A case report. AB - BACKGROUND: Coexistence of cancer and tuberculosis in axillary lymph nodes is rare. Only seven cases have been reported in the literature. CASE REPORT: We report here a case of infiltrating ductal carcinoma breast metastasizing to the axillary lymph node along with tubercular granuloma in the same lymph node without primary mammary or pulmonary tuberculosis. CONCLUSION: Primary tuberculosis coexisting with carcinoma is of rare occurrence. A possibility should always be borne in mind especially in patients from endemic areas. PMID- 12773223 TI - Pleomorphic adenoma of minor salivary gland in the parapharyngeal space. AB - BACKGROUND: Majority of tumours occurring in minor salivary gland are malignant. Pleomorphic adenoma is of rare occurrence in parapharangeal space. CASE PRESENTATION: A rare case of a minor salivary gland pleomorphic adenoma of the parapharyngeal space is reported. Review of literature, clinical features, pathology, radiological findings and treatment of these tumours are discussed. CONCLUSIONS: Parapharangeal salivary tumours presents as a painless progressive swelling with majority of these being malignant. Pleomorphic adenoma in parapharangeal space is of rare occurrence. Complete surgical excision is the treatment of choice. PMID- 12773224 TI - Festschrift in honour of Dr. Ernest Ramsey. PMID- 12773225 TI - An integral view of the neuroendocrine aspects of male sexual dysfunction and aging. AB - Age is the most important factor associated with sexual dysfunction. The traditional thinking explained this association by the neurovascular events developing with the aging process. More recently, central neuro-endocrine mechanisms resulting from apoptosis in the hypothalamic areas involved in both the production of sex hormones and the control of sexual processes have added a new dimension to human sexuality. This evidence is still controversial but of unquestionable importance in our understanding of endocrine alterations and sexual shortcomings associated with aging. PMID- 12773226 TI - A comparison of extracorporeal shock wave lithotripsy and ureteroscopy under intravenous sedation for the management of distal ureteric calculi. AB - INTRODUCTION: We have performed a study to compare shock wave lithotripsy (SWL) and ureteroscopy under intravenous sedation for the management of distal ureteric calculi. MATERIALS AND METHODS: Patient tolerance, procedure times and treatment outcomes were prospectively evaluated in 110 patients undergoing 138 SWL treatments, and 172 patients undergoing ureteroscopy under intravenous sedation for the management of distal ureteric calculi. RESULTS: Men tolerated SWL better than ureteroscopy. Over 90% of women tolerated both procedures well. Procedure times were 52 minutes for SWL and 27 minutes for ureteroscopy. Treatment was successful in 72% of patients undergoing SWL, and 95% of patients undergoing ureteroscopy. CONCLUSIONS: In women with distal ureteric calculi requiring treatment, we recommend ureteroscopy under intravenous sedation as the treatment of first choice. In men the better tolerance of SWL must be weighed against the higher success rate of ureteroscopy. If both treatment modalities are available, patients with small distal ureteric calculi, in whom ureteroscopy is likely to be successful, should be informed of and offered their choice of either treatment modality. PMID- 12773227 TI - Probiotics and the urologist. AB - Emerging from the stigma of once being referred to as "snake oil", excellent scientific and clinical evidence now exists to indicate that probiotics do indeed have a role to play in medicine. The proper definition of probiotics is important "Live microorganisms which when administered in adequate amounts confer a health benefit on the host", for several reasons. It rules out so-called probiotics that have no clinically proven, peer-reviewed data, and it states the need to have viable bacteria present, unlike these pseudo products which are often wrongly labeled, poorly manufactured, with low or no viability at time of use. Guidelines, prepared by the United Nations and World Health Organization are now available to guide physicians and consumers as to the types of strains with documented benefits. In urology, the most studied strains are Lactobacillus rhamnosus GR-1 and L. fermentum B-54 and RC-14. Their use daily in oral form, or once to three times weekly as a vaginal suppository, have been shown to reduce the urogenital pathogen load and the risk of urinary tract and vaginal infections. Organisms such as Oxalobacter formigenes, still in the R&D phase, offer great potential to reduce kidney stone formation via oxalate degradation in the intestine. Some studies using L. casei Shirota suggest a possible effect against bladder cancer, while studies using L. plantarum 299 show significantly reduced infection rates in patients undergoing major surgical procedures. In short, specific probiotic strains hold much promise for use in the urology setting. PMID- 12773228 TI - Management of bacillus Calmette-Guerin (BCG) refractory superficial bladder cancer: results with intravesical BCG and Interferon combination therapy. AB - INTRODUCTION AND OBJECTIVE: BCG is the most efficacious intravesical treatment for superficial bladder cancer. However, 30%-40% of tumors are refractory. BCG failure is an indication for cystectomy but several salvage intravesical (IVe) strategies have been proposed. Early results with reduced dose BCG in combination with IFN-a in patients are currently the most promising. We have adopted this approach and now report our preliminary results. This is the first report of this salvage therapy from Canada, the birthplace of IVe BCG therapy for superficial bladder cancer. METHODS: The "O'Donnell protocol" of reduced dose IVe BCG plus IFN-a was followed in 12 patients with BCG refractory superficial transitional cell carcinoma. A retrospective review of the efficacy and toxicity of the treatment was conducted. RESULTS: One year from induction therapy with salvage BCG/IFN-a, 6 of the 12 (50%) of patients were tumor free. Of the six recurrences, 3(50%) did not respond to the IVe therapy and had residual/recurrent tumor at the first follow-up visit. Risk factors for treatment failure were identified. The combinative therapy was well tolerated with minimal toxicity compared to previous full dose BCG. CONCLUSION: Our 12 month data with reduced dose IVe BCG plus IFN-a salvage therapy for BCG refractory superficial TCC confirm previous reports of >50% complete response rates. We need longer follow up in a larger patient population to determine the durability of this promising therapy in patients who would otherwise undergo radical cystectomy. PMID- 12773229 TI - Apoptosis in the prostate. AB - The prostate requires androgens for development and glandular maintenance, dying by the process of apoptosis following their removal. Anti-androgen therapy is targeted to induce this process but eventually fails with the emergence of an androgen independent cancer. These cells have development mechanisms to survive with out androgen impart due to the expression of anti-apoptotic factors. PMID- 12773230 TI - Benign prostatic hyperplasia: from A - Z. AB - The management of lower urinary tract symptoms (LUTS) secondary to benign prostatic hyperplasia over the last decade underwent many changes. The introduction of many medical options including alpha blockers and 5 alpha reductase inhibitors provided alternatives to what used to be surgery or "watchful waiting". Alpha blockers evolved over the years from non specific alpha 1, and 2 blockers to alpha 1 selective and then to alpha 1a selective with a wider acceptance due to lack of need to titrate and a better safety profile. 5 alpha reductase inhibitor (finasteride) passed through a lot of changes from being the first medication directed at treating the disease process to less acceptance because of time to response and early data supporting no added benefit when combined to alpha blockers for a short period. Longer studies now demonstrate a benefit to combination causing a reduction of progression parameters and an advantage over 4 years in reducing endpoints, namely acute urinary retention and surgery. Surgical options have also undergone many changes over the last decade with introduction of minimally invasive options as well as the introduction of new energy sources to reduce complications and allow for management of larger glands such as Holmium laser enucleation of the prostate or the use of bipolar loops. The journey has been long and exciting and we are sure Ernie Ramsey enjoyed being in the forefront of the evolution. PMID- 12773231 TI - Clinical impact of adjunctive donor microvascular reconstruction on renal transplantation. AB - INTRODUCTION: Microvascular reconstruction was incorporated into our donor organ harvesting algorithm for kidneys with anatomic anomalies or injury of the vasculature. The impact of adjunctive microsurgery was appraised in terms of organ availability and graft quality procedures. METHODS: Out of a total of 441 renal transplant procedures performed by one surgeon (JLC) between 1984 and 1997, 104 allografts (83 cadaveric, 21 living related) required ex-vivo microvascular reconstruction. Micro-reconstruction using 2.5-10 X magnification was employed to create a single artery and vein for subsequent in-situ anastomosis. Side-to-side or end-to-side anastomosis was performed, depending on the vascular arrangement. Multiple vessels and those injured during harvesting were reconstructed with a combination of the above techniques. RESULTS: Eleven kidneys had two or more arterial anastomoses; 12 had combination (arterial and venous) anastomoses while 74 required a single micro-reconstruction. In addition, seven kidneys with severely traumatized vessels were salvaged. Average bench surgery times were 30 and 50 minutes for single and multiple reconstructions respectively. Mean warm ischemic time was 29 minutes. Three kidneys were lost due to vascular thrombosis (two venous, one arterial) where in-situ technical difficulties were encountered in all three cases. With mean follow-up of 30 months, 23 kidneys had been lost due to chronic rejection with the remainder functioning. CONCLUSION: Extensive microvascular reconstruction salvaged 30 suboptimal or previously deemed unusable grafts (30/439 = 7%) and facilitated the vascular anastomosis in another 74 cases (17%). The warm ischemic time and the possibility of in-situ technical errors with small-calibre vessels were minimized. This report affirms the contention that microvascular reconstruction should be available as an adjunctive technique for renal transplantation, to maximize the quantity and quality of donor kidneys. PMID- 12773232 TI - Intermittent androgen suppression in prostate cancer: an update of the Vancouver experience. AB - INTRODUCTION: This report will review the long-term follow-up of a prospective Phase II evaluation of intermittent androgen suppression in the treatment of prostate cancer. Specifically, this analysis will address completed cycle characteristics, the concept of prolonged off-treatment cycles, the time to cancer progression, cancer-specific survival and the association between PSA and bone scan changes. METHODS: A total of 102 patients have been entered into this protocol. Treatment was initiated with combined androgen blockade and continued for 6 months or longer to reach a serum PSA nadir. Medication was then withheld until the serum PSA increased to predetermined trigger points based on initial parameters. Each cycle of treatment and no-treatment was repeated until the regulation of PSA became biochemically androgen independent. RESULTS: One hundred two patients have been commenced on IAS with an average follow-up time of 219 weeks (range: 14.5 to 588). Ninety-one patients have completed at least one therapeutic cycle with a total of 188 completed cycles available for analysis. The average time off therapy (percentage time off therapy) for cycles 1, 2, 3 and 4 was 13 months (53%), 11 months (51%), 10 months (47%) and 8 months (45%), respectively. A prolonged off-treatment time of greater than 72 weeks was observed in 33 (18%) of all completed cycles, and was most common in the men being treated for radiation failure stage C. Progression and survival data was calculated for the entire trial cohort (n=102). The average time to androgen independent progression in the 29 (28%) patients who progressed was 194 weeks. Death from prostate cancer occurred in 19 (18%) patients at an average of 258 weeks following treatment initiation. A review of bone scans revealed 22 events of newly detected lesions, all but 2 of which were preceded by a rise in serum PSA. CONCLUSIONS: Longer duration follow-up of a single cohort continues to support IAS as a viable treatment option for men with prostate cancer. This approach affords an improved quality of life when the patient is off therapy, with reduced toxicity and costs. There is a trend toward extended times to progression and death compared to contemporary studies of continuous androgen suppression. Randomized, prospective protocols are currently underway to determine whether survival is affected in a beneficial or adverse way in men with locally recurrent or metastatic cancer. PMID- 12773233 TI - Genitourinary tract preservation versus pelvic exenteration for advanced colorectal tumors. AB - INTRODUCTION: Total pelvic exenteration (TPE) is the standard of care for locally advanced colorectal cancer. This is a morbid procedure often leaving the patient with two ostomy sites and an extended recovery. Bladder preservation with complete tumor resection is often possible in these cases and we set out to determine if limited resection of the GU tract was as effective in tumor control as TPE. MATERIALS AND METHODS: This is a retrospective review of all patients over a 7-year period with colorectal tumors invading the urinary system. These patients were divided into two groups based upon the surgical procedure they received: TPE or GU tract sparing surgery. Tumor stage, adjuvant cancer therapy, and complications were reviewed. Recurrence and survival rates were calculated. RESULTS: There were 19 patients with colorectal tumors invading the GU tract. Eight patients were treated with TPE and 11 patients received GU tract sparing procedures. Tumor stage and extent of disease were similar for both groups as were adjuvant/neoadjuvant therapy received. The average follow up from surgery for TPE and GU tract sparing procedures was 40 months (range 9 - 96) and 53 months (range 21.5 - 94), respectively. The limited resection and TPE groups experienced similar complication rates: 7/11 (63.6%) and 6/8 (75%) respectively. The 5-year survival rate was 37.5% and 61.4% for TPE and limited GU resections, respectively (p=0.07). CONCLUSION: The cancer recurrence and complication rates were similar in both groups offering no clear advantage to TPE. GU sparing surgery in the face of locally invasive colorectal tumors is a viable option allowing for treatment of the disease and reducing the morbidity of total pelvic exenteration. PMID- 12773234 TI - [The latest consensus for hepatic encephalopathy]. PMID- 12773235 TI - [Hepatitis B virus genotypes and the heterogeneity of its polymerase gene]. AB - OBJECTIVE: To study the heterogeneity of polymerase gene (P gene) within hepatitis B virus (HBV) genotypes based on a systematic analysis of 202 HBV P genes, providing some useful references for further studies on the relationship among HBV genotypes, P gene mutations, replication and nucleoside analogues drug resistance. METHODS: 202 HBV complete sequences containing P genes were obtained from GenBank and were analysed using computer softwares. RESULTS: There were some genotype-related characteristics of HBV P genes. As reverse transcriptase domain was concerned, there were more amino acid divergences in genotype C and D compared with these in genotype A. There were also amino acid substitutions in the A-F conserved regions of the reverse transcriptase domain within and between HBV genotypes. CONCLUSIONS: There are divergences of P genes and amino acids within and between HBV genotypes, which should be considered when amino acid changes are analyzed whether they are proposed to be drug-resistance mutations or the results from quasispecies-selected. Moreover, these divergences may affect the antiviral effect of nucleoside analogues on HBV with different genotypes. PMID- 12773236 TI - [Effect and mechanism of beta-L-D4A (a novel nucleoside analog) against hepatitis B virus]. AB - OBJECTIVE: To explore the effect and the molecular targets of anti-hepatitis B virus (HBV) by beta-L-D4A in vitro. METHODS: 2.2.15 cells were cultured and treated with various concentrations of beta-L-D4A for 6 hours, then the effect of anti-HBV was examined by Southern blot and the replicating core particles from the cells were isolated. The endogenous polymerase reaction and activity gel experiment were performed to monitor the activities of the DNA polymerase and reverse transcriptase. RESULTS: The replication of HBV DNA was inhibited in a dose-dependent manner. The endogenous polymerase reaction showed both the two enzymatic activities were irreversibly inactivated in a concentration -dependent manner, with IC50 at 0.51 micromol/L and 0.55 micromol/L, respectively. But the activities of DNA polymerase and reverse transcriptase were found to remain active by activity gel with exogenous templates. CONCLUSIONS: The mechanism of inhibiting HBV replication by beta-L-D4A may be in that either the DNA replication priming is blocked or the elongation of DNA chain is terminated irreversibly. PMID- 12773237 TI - [Heat shock protein 70-HBsAg complex inducing antigen-specific cytotoxic T lymphocyte immune response]. AB - OBJECTIVE: To explore the possibility of cell-medicated immune response induced with heat shock protein 70 (HSP70)-HBsAg protein complex in vitro. METHODS: HSP70 HBsAg complex was reconstituted in vitro which was injected into mice in order to observe that whether HSP70-HBsAg would stimulate humoral and cellular immune responses. HSP70, HSP70-HBsAg complex and HBsAg were used to activate the dentritic cell (DC) individually, which would initiate homogeneic T lymphocyte to transform to cytotoxic T lymphocyte (CTL). The cytotoxicity of CTL was detected with MTT assay. RESULTS: HSP70-HBsAg complex elicited both humoral and cellular immune responses against HBsAg in mice. Specific CD8+ CTL response was readily induced by HSP70-HBsAg complex and HBsAg, especially the former. CONCLUSIONS: HSP70-HBsAg complex is immunogenic and HSP70 can lead to great efficient CTL response. And HSP70-HBsAg complex may be used as a protein vaccine for immunotherapy for chronic hepatitis B. PMID- 12773238 TI - [Identifying the gene encoding hALR-associated proteins using yeast two-hybrid system]. PMID- 12773239 TI - [Long-term health-related quality of life in chronic hepatitis B patients]. AB - OBJECTIVE: To evaluate their long-term outcome and the efficacy and economic significance of antiviral drugs by investigating the long-term health-related quality of life (HQL) in chronic hepatitis B (CHB) patients. METHODS: The HQL of 101 CHB patients with biopsy-proven 6 to 18 years ago and 105 persons of general population as control was studied with revised SF-36 questionnaire. RESULTS: The HQL in CHB patients was lower than that in general population in physical functioning, role physical, general health, mental health, and specific symptoms (mu > or = 2.10, P<0.05). CONCLUSIONS: The long-term HQL in chronic hepatitis B patients is poor. PMID- 12773240 TI - [Expression and clinical significance of p27kipl and Rb proteins in primary liver cancer]. PMID- 12773241 TI - [Effect of ribozyme against platelet-derived growth factor receptor beta subunit mRNA on the biological characters of hepatic stellate cells]. AB - OBJECTIVE: To study the cleavage activity of hammerhead ribozyme targeting at platelet-derived growth factor receptor beta subunit (PDGFR- beta) mRNA in hepatic stellate cells (HSCs) and its effect on the biological characters of HSCs. METHODS: Expression vector of anti-PDGFR- beta ribozyme was constructed and transfected into rat-derived HSC-T6 cells with lipofectin. The positive cell clones were gained by G418 selection. The expression of PDGFR- beta, alpha-smooth muscle actin (alpha-SMA), and type I and type III collagen was detected by means of northern blot, Western blot and immunocytochemical staining respectively. The cell proliferation was determined with MTT colorimetric assay. The cell apoptosis was demonstrated with flow cytometry, acridine orange fluorescence vital staining and transmission electron microscopy. RESULTS: The expression of PDGFR- beta at mRNA and protein level was markedly reduced in ribozyme-transfected HSCs only 43% to 51% of that in control cells (t > or = 3.957, P < 0.05), and alpha-SMA expression level, type I and type III collagen synthesis ability were also reduced (t > or = 6.790, P < 0.01). The proliferation of ribozyme-transfected HSCs was significantly decreased (t > or = 3.858, P < 0.05), and the proliferation response to PDGF BB was markedly inhibited. However the apoptotic rate was significantly increased in ribozyme-transfected HSCs (chi2 > or = 14.157, P < 0.01), and typical apoptotic cells could be found under transmission electron microscopy. CONCLUSIONS: The anti-PDGFR- beta ribozyme can be expressed stably in HSCs, cleave the target RNA effectively, inhibit HSCs proliferation and collagen synthesis, and induce HSC apoptosis. The results suggest that inhibiting PDGFR- beta expression in HSCs may be a new therapy for liver fibrosis. PMID- 12773242 TI - [Effect of blocking transforming growth factor beta signalling on culture activated rat hepatic stellate cells]. AB - OBJECTIVE: To study the effects of blocking transforming growth factor beta (TGF beta) signalling on culture-activated rat hepatic stellate cells (HSCs). METHODS: After cultured in plastic dish for two days, HSCs were infected with adenovirus vector AdT beta-ExR or AdLacZ (control) at 10 multiplicity of infection (MOI) and incubated for four days. The expression of type I collagen, alpha-smooth muscle actin (alpha-SMA) and the proliferation of HSCs were analyzed by ELISA, western blot, immunocytochemistry and BrdU uptake respectively. RESULTS: The expression level of type I collagen in HSCs infected with AdT beta-ExR was 42.99% of that in HSCs infected with AdLacZ (q = 9.100, P < 0.001). The expression of alpha-SMA in HSCs infected with AdTbeta-ExR was also inhibited evidently. But the BrdU uptake in HSCs infected with AdLacZ was 49.24% of that in HSCs infected with AdTbeta-ExR (q = 7.835, P < 0.001). CONCLUSIONS: The blockade of TGF-beta signalling in cultured rat HSCs can inhibit their activation significantly, but promote their proliferation. PMID- 12773243 TI - [Effect of HanDanGanLe on the cytokines in fibrotic rats]. AB - OBJECTIVE: To study the expression of connective tissue growth factor (CTGF) mRNA and transforming growth factor beta 1 (TGFbeta1) mRNA in immunity-induced liver fibrosis rats and the effect of HanDanGanLe on them. METHODS: Male wistar rats were given intraperitoneal injection of porcine serum twice a week. At the beginning, the rats in HanDanGanLe-treatment group were feed with HanDanGanLe for precaution. The rats were killed after twelve weeks, then CTGF mRNA and TGFbeta1 mRNA were detected in liver samples with in situ hybridization, and the formation of liver fibrosis was observed with HE stain. The semi-quantitative RESULTS: of the two genes expression were analysed along with the stages of hepatic fibrosis. RESULTS: Typical liver fibrosis developed in the model group rats, and the positive stain of CTGF mRNA and TGFbeta1 mRNA increased, which was distributed in the areas where fibrosis occurred. There was obvious correlation between the expression strength of CTGF mRNA and TGFbeta1 mRNA (r = 0.799, P < 0.05). In the rats receiving HanDanGanLe, CTGF mRNA expression index decreased markedly (12.5+/ 2.3 vs 28.8+/-1.4, t = 5.208, P < 0.01), so did TGFbeta1 mRNA expression index (25.4+/-3.2 vs 37.3+/-5.4, t = 5.655, P < 0.01). There was also significant correlation between the scores of CTGF mRNA expression and the stages of hepatic fibrosis (rs = 0.822, 0.808 in the model group and HanDanGanLe-treatment group, P < 0.05). CONCLUSIONS: The expression of CTGF mRNA and TGFbeta1 mRNA is correlated closely with hepatic fibrosis degree. HanDanGanLe can effectively prevent the expression of CTGF and TGFbeta1. One of the mechanisms of the intervention may be its blocking the intracellular signalling pathways involved in liver fibrogenesis. PMID- 12773244 TI - [Effect and mechanism of Tanshensu on fibrotic rats]. AB - OBJECTIVE: To investigate the effect and mechanism of Tanshensu on experimental fibrotic rats. METHODS: Pigs serum was used to induce liver fibrosis in Wistar rats. The rats in Tanshensu-treated group were injected peritoneally with Tanshensu solution at the dose of 300 mg x kg(-1) x d(-1), and the rats in model control group and normal-control group received the same volume of double distill water. At the end of the twelfth week, the hepatic stellate cells (HSCs) were isolated from the liver of one rat in model-control group using in-situ perfusion with pronase and collagenase, then density gradient centrifugation, and the other rats were killed to take the serum and liver samples. MTT colorimetric assay was used for detecting the proliferation of HSCs and flow cytometry was used for observing the cell cycles of HSCs under different concentrations of Tanshensu. The hyaluronic acid (HA) level in serum was detected and the morphological changes of liver tissue were observed. RESULTS: There was a decline of serum HA level in Tanshensu-treated group compared to that of the model-control group (231.4 ng/ml +/- 41.1 ng/ml vs. 398.7 ng/ml +/- 54.5 ng/ml, F =154.796, P < 0.05). Both HE and VG stain showed a decline of liver fibrosis degree in Tanshensu-treated group. And Tanshensu had an inhibition effect on the proliferation of HSCs at the concentrations of 50 mg/L, 100 mg/L, and 200 mg/L (1.60x10(-2) +/- 8.17x10(-4), 1.10x10(-2) +/- 1.41x10(-3), and 6.75x10(-3) +/- 3.30x10(-3) vs. 7.18x10(-2) +/- 1.71x10(-3), F =1154.221, P <0.01). CONCLUSIONS: Tanshensu shows a therapeutic effect on liver fibrosis in rats induced by pig's serum through inhibiting the proliferation of hepatic stellate cells. PMID- 12773245 TI - [Inhibition of HBV DNA replication and expression in 2.2.15 hepatoma cells infected with AFP-mediated HBX antisense RNA]. AB - OBJECTIVE: To study the specific expression of the antisense RNA against hepatitis B virus X (HBX) gene in hepatoblastoma cell line and its anti -HBV activity. METHODS: HBX gene (nt.1370-1827) was amplified by PCR, then cloned into EB virus vector pEBAF which contained human alpha-fetoprotein promoter and enhancer. After transfected into 2.2.15 hepatoma cells and ECV304 human endothelial cells by lipofectin, northern blot, ELISA and real-time qualitative PCR were carried out to assay the expression of HBX mRNA, HBV antigens and HBV DNA level, respectively. RESULTS: The HBX antisense RNA expression vector pEBAF as-HBX which could be expressed specifically in 2.2.15 hepatoblastoma cells was successfully constructed. Both HBV DNA level and the expressions of hepatitis B virus surface antigen (HBsAg) and e antigen (HBeAg) in 2.2.15 hepatoblastoma cells were inhibited by pEBAF-as-HBX. Compared with those in sense control (pEBAF s-HBX), the inhibitory rates of HBsAg, HBeAg, and HBV DNA were 37.9%, 36.8%, and 25%, respectively. CONCLUSIONS: The pEBAF-as-HBX expression vector may lead to targeted-expression of HBX antisense RNA in hepatoma cells and shows great inhibition effect on HBV. PMID- 12773246 TI - [Values of soluble thrombomodulin and von Willebrand factor judging reject reaction in liver transplantation]. AB - OBJECTIVE: To find sensitive and specific laboratory examination items for early diagnosing and monitoring liver transplantation reject reaction. METHODS: Randomly investigate 41 liver transplantation patients, among them there were 16 patients with reject reaction (including 12 with acute rejection, 4 with chronic rejection). Plasma soluble thrombomodulin (STM) and von Willebrand factor (vWF) levels were measured before operation and every other day after operation. RESULTS: Plasma STM level increased significantly after operation, two days before rejection and after acute rejection (5.58 ng/ml +/- 0.42 ng/ml, 5.93 ng/ml +/- 0.45 ng/ml, and 7.88 ng/ml +/- 0.29 ng/ml, respectively), so did vWF level (101.2% +/- 4.68%, 104.3% +/- 5.78%, and 127.7% +/- 5.74%, respectively). STM level was much higher in acute rejection than that in chronic rejection (7.88 ng/ml +/- 0.29 ng/ml vs. 6.35 ng/ml +/- 0.54 ng/ml, t = 2.46, P < 0.05), in no reaction group after impacting therapy than in effective group (8.30 ng/ml +/- 0.19 ng/ml vs. 3.82 ng/ml +/- 0.22 ng/ml, t = 12.98, P < 0.01), and in dead group after treatment than in living group (7.98 ng/ml +/- 0.18 ng/ml vs. 6.51 ng/ml +/ 0.41 ng/ml, t = 3.39, P < 0.01). CONCLUSIONS: Plasma STM and vWF can be taken as laboratory items for monitoring liver transplantation rejection. Plasma STM can act as not only an early prognosticating marker, but also suitable to distinguish acute from chronic reject reaction, and as a marker for monitoring impacting therapy effect and judging prognosis. PMID- 12773247 TI - [Effect of Tauroursodeoxycholic acid on cytochrome C-mediated apoptosis in HepG2 cells]. AB - OBJECTIVE: To investigate the effect of Tauroursodeoxycholic acid (TUDCA) on Taurodeoxycholic acid (TDCA)-induced HepG2 cell apoptosis and to clarify the molecular mechanism of its anti-apoptosis effect of TUDCA. METHODS: Morphologic evaluation of apoptotic cells was performed by Hoechst 33258 staining and electron microscope. DNA fragment was detected by electrophoresis on 1.5% agarose gels. Apoptosis rate was measured by flow cytometry using PI dye. Following incubation of HepG2 cells either with TDCA alone, or coincubation with TUDCA and TDCA, the releasing level of cytochrome c from mitochondria into cytosol was determined by western blot, also the activity of caspase-3, 8, 9. RESULTS: Incubating the cells with 400 micromol/L TDCA for 12 h induced the cells apoptosis significantly. The apoptotic rate decreased from 50.35% +/- 2.20% to 13.78% +/- 0.84% after coincubation with TUDCA, and this anti-apoptotic effect of TUDCA was confirmed by morphological and DNA ladder detection. TUDCA significantly inhibited the release of cytochrome C from mitochondria into cytosol, and the activity of caspase-9, 3 (t > or = 13.00, P < 0.01), especially at 12 h, caspase-3 activity decreased by 54.9% (t = 16.88, P < 0.01) and 52.5%, however it had no obvious effect on the activity of caspase-8 (t = 1.94, P > 0.05). CONCLUSIONS: TUDCA prevents HepG2 cells apoptosis induced by TDCA through modulating mitochondrial membrane stability, inhibiting the release of cytochrome c and the activation of procaspase-9 and 3. Anti-apoptotic mechanism of TUDCA may be considered to be one of the most important reasons that TUDCA exerts significant efficacy in the treatment of cholestatic liver diseases. PMID- 12773248 TI - [Types and emergence time of YMDD motif mutation in hepatitis B virus polymerase gene during lamivudine treatment]. AB - OBJECTIVE: To study the types and emergence time of YMDD motif mutation in hepatitis B virus (HBV) polymerase gene during lamivudine treatment. METHODS: The serum samples were collected from 33 patients with HBV DNA rebounding and 2 non responders after at least one year lamivudine treatment. HBV polymerase gene was amplificated by PCR, then the products were detected by restriction fragment length polymorphism (RFLP) and by direct sequence analysis. RESULTS: The variants with YMDD mutation were 14 out of the 35 patients. Mutation patterns detected in these patients included four YIDD, six YVDD, three YI/VDD and one YI/MDD. The mean emergence time of YMDD variants was 11.07+/-3.65 months after the treatment, and the earliest one and the latest one occurred 5 months and 17 months after the treatment respectively. The emergence times of YIDD, YVDD, YI/VDD were (10.00 +/- 1.41) months, (11.67 +/- 4.41) months and (13.33 +/- 3.31) months respectively, which had no statistical significance (F = 0.543, P < 0.05). Three patients treated with lamivudine 200 mg every day after the mutation were followed up for 6 months, whose HBV variants had not vanished. CONCLUSIONS: There are many kinds of HBV variants after lamivudine treatment, including YIDD, YVDD, YI/VDD and YI/MDD. The emergence time of variants is quite variable between different types and the mean time is (11.07 +/- 3.65) months after treatment, and there is no relationship between the type of YMDD mutation and the time of lamivudine administration. PMID- 12773249 TI - [Therapy effect of lamivudine combination with alpha interferon on patients with chronic hepatitis B]. AB - OBJECTIVE: To evaluate the short-term therapeutic efficacy and safety of lamivudine (LAM) combining with alpha interferon (IFNalpha) on patients with chronic hepatitis B. METHODS: 90 chronic hepatitis B patients with HBV DNA and HBeAg positive were subdivided by 1:1:1 proportion into three groups: (1) LAM+IFN group: 6 months therapy of IFNalpha plus lamivudine followed by 6 months of lamivudine; (2) LAM group: lamivudine alone for 12 months; (3)IFN group: IFNalpha alone for 6 months. RESULTS: At the end of treatment, the HBV DNA undetectable rate in LAM+IFN group (90.0%) was much higher than that in LAM group (80.0%) and IFN group (46.7%) (chi2 = 13.017, P < 0.001). ALT normalization occurred 90.0%, 80.0%, and 53.3% in LAM+IFN group, LAM group, and IFN group, respectively (chi2 = 9.932, P = 0.002). HBeAg/anti-HBe seroconversion rates achieved 46.7%, 13.3%, and 33.3% in LAM+IFN group, LAM group, and IFN group, respectively (chi2 = 7.937, P = 0.005). YMDD mutation was not detected in serum samples from LAM+IFN group patients. CONCLUSIONS: LAM+IFN therapy for chronic hepatitis B is tolerated and more effective than IFN monotherapy in inhibiting viral replication and getting ALT normalization. The HBeAg/anti-HBe seroconversion rate with LAM+IFN therapy is higher than that with lamivudine monotherapy. LAM+IFN combination therapy seems to inhibit or postpone YMDD variants appearing in patients with chronic hepatitis B. PMID- 12773250 TI - [Effects of herbal compound 861 on hepatic stellate cell expressing endothelin-1 protein and mRNA]. PMID- 12773251 TI - [Effects of antisense oligonuclotide on c-myb expression in rats with liver fibrosis]. PMID- 12773252 TI - [Establishment and application of a genotyping method for hepatitis B virus using polymerase chain reaction]. PMID- 12773253 TI - [Application of acute physiology and chronic health evaluation II scoring system in severe hepatitis]. PMID- 12773254 TI - [Changes and significance of tumor necrosis factor-alpha and interleukin-6 level in plasma and bile during the formation of acute intrahepatic cholestasis in New Zealand white rabbits]. PMID- 12773255 TI - [Role of caspase-8 and caspase-3 in hepatoma cells apoptosis induced by 5 fluorouracil]. PMID- 12773257 TI - [Retrospect and progress of hepatopulmonary syndrome]. PMID- 12773256 TI - [Smad 7 and post-receptor signal regulation of transforming growth factor-beta1]. PMID- 12773258 TI - Early thrombolysis for the treatment of acute myocardial infarction: a systematic review and economic evaluation. PMID- 12773259 TI - Evaluation of molecular tests for prenatal diagnosis of chromosome abnormalities. PMID- 12773260 TI - Systematic review of the effectiveness and cost-effectiveness, and economic evaluation, of home versus hospital or satellite unit haemodialysis for people with end-stage renal failure. PMID- 12773261 TI - Effect of naphthalan on epidermal proliferation activity and CD3, CD4, and CD8 lymphocyte count. AB - Immunohistochemistry analysis by cell immunophenotyping before and after a 3 - week treatment with naphthalan oil was performed on biopsy specimens from 10 patients with psoriasis vulgaris. For immunohistochemistry staining of 3- micro m paraffin block sections, anti-CD3, anti-CD4, anti-CD8, and Ki-67 antibodies were used. Peroxidase reaction for T cell (CD3, CD4, and CD8) quantification was done in the epidermis and dermis of each skin sample as total positive cell count per mm sample. Positive Ki-67 (proliferation index) was determined as percentage of positive cells per 100 cells in the basal layer of the epidermis. Results showed naphthalan treatment to decrease the mean CD3 lymphocyte count by 83% and 59%, CD4 lymphocyte count by 81% and 73%, and CD8 lymphocyte count by 60% and 49% in the epidermis and dermis, respectively. The mean proliferation index also decreased with naphthalan therapy. PMID- 12773262 TI - Correlation of clinical symptoms and laboratory findings in children with milk allergy. AB - Food hypersensitivities can be divided into toxic and nontoxic, and the latter can further be subdivided into immune and nonimmune hypersensitivities. Cow's milk allergy or intolerance occurs in 5-15% of infants, mostly during the first year of life, or occasionally later. The symptoms may involve different organ systems, especially the gastrointestinal system, skin, and respiratory system. For the diagnosis of cow's milk protein allergy/intolerance, double-blind, placebo-controlled food challenge has been used as a gold standard. Since the test suffers from some drawbacks, many reports have pointed to the need for novel and simpler diagnostic procedures and criteria. In our study, clinical symptoms and laboratory findings of patients with cow's milk protein allergy were compared to assess the possible correlation between particular laboratory findings, clinical picture, and the organ system predominantly involved. There were no significant differences in the levels of IgE, cow's milk protein specific IgE, eosinophilia, prick test results, rectal mucosa biopsy histology, and atopy incidence in patient families among the children with gastrointestinal, cutaneous, and combined gastrointestinal and cutaneous symptoms. Improvement in the symptoms with dietary therapy irrespective of clinical presentation and type of hypersensitivity underlying the symptoms in all these patients strongly suggests that clinical response should be a basic criterion for the diagnosis of cow's milk protein allergy. PMID- 12773263 TI - Frequency of standard and occupational contact allergens in Tuzla area, Bosnia and Herzegovina: retrospective study. AB - Allergic contact dermatitis is acute or chronic inflammatory skin disease of allergic etiology, which develops as a result of delayed type of hypersensitivity, i.e. type IV reaction according to the Gell and Coombs classification. In the retrospective study, we reviewed medical records of 495 patients diagnosed with allergic contact dermatitis in the 1988-1998 period. The records were obtained from the Cabinet of Allergology of the Department of Dermatology and Venerology, Tuzla University Hospital Center. There were 312 women and 183 men, aged between 18 and 60 years. The patients were divided into 6 groups according to their occupation. Contact sensitization was established in 295 or 59.6% of them. The type and frequency of causative agent in allergic contact dermatitis depended on working environment. Potassium dichromate, a component part of cement, caused positive reaction in 48% of construction workers tested, which was significantly more than in the "other occupation" group, where 14.6% of patients showed positive reaction to potassium dichromate (p<0.001). Formaldehyde, used in leather processing, was the most frequent among the four leading allergens in the group of shoe workers (13.3%), whereas charcoal tar (used in the metal processing) was the most frequent allergen in the group of metal workers (13.9%). Nickel sulfate, potassium dichromate, cobalt chloride, and urushiol were frequent allergens in the "other occupation" group, housewives, and textile workers. The listed allergens are present at large in everyday life as well as in particular occupations. PMID- 12773264 TI - The use of emollients as sophisticated therapy in dermatology. AB - Emollients are agents designed to make the stratum corneum softer and more plant by increasing its hydration. A large number of preparations are available today, many of which are marketed as cosmetic and therapeutic moisturizers. They are the most prescribed products in dermatology. Their structure and function are surprisingly complex and sophisticated, and many are equidistant between cosmetics and drugs. The use of the emollients corrects the problems in scaling disorders. It is well known that the electrical properties of the stratum corneum change after application of an emollient. It is also possible that they have suppressive effects on epidermal thickening. Emollients have an anti-inflammatory activity and also give some transient relief from irritation. In clinical use emollients are employed as treatments for ichthyoses, xeroderma and disorders of keratinization, atopic dermatitis, psoriasis, and photodamaged skin. Emollients of the new millennium include agents that mimic natural ingredients and function as botanicals, including vitamins, hydroxy acids, and retinoids. Emollients can cause a few side effects, such as irritant dermatitis, allergic contact dermatitis, fragrance allergy or allergy to other constituents (preservatives or additives), stinging, cosmetic acne, and pigmentary disorders. We can conclude that emollients, continuously evolving to ever more sophisticated products, are very important in the treatment of different dermatoses. PMID- 12773266 TI - Risperidone and borderline personality disorder. PMID- 12773265 TI - Thyroid hormone and mood modulation: new insights from functional brain imaging techniques. PMID- 12773268 TI - Selective serotonin reuptake inhibitors in women with borderline personality disorder. PMID- 12773270 TI - Chronic hallucinatory psychosis, bouffee delirante, and the classification of psychosis in French psychiatry. AB - The conflict between French nosology and international classifications is mainly linked to the French concepts of chronic hallucinatory psychosis and bouffee delirante. However, these discrepancies are now largely reduced by the evolution of the recent versions of international classifications. The term chronic hallucinatory psychosis is used to describe a chronic hallucinatory and delusional disorder that differs from paranoid schizophrenia in the absence of formal thought disorder and intellectual impairment. This concept appears to be quite similar to paranoid schizophrenia as defined by the Diagnostic and Statistical Manual of Mental Disorders (DSM) and International Classification of Disease (ICD). However, the recent statement that deficit and nondeficit schizophrenia are separate diseases rediscovered French distinctions between chronic hallucinatory psychosis and schizophrenia. The term bouffee delirante describes an acute nonaffective and nonschizophrenic psychotic disorder, which is largely similar to DSM-III-R and DSM-IV brief psychotic and schizophreniform disorders, and was taken up in ICD-10 under the name acute polymorphic psychotic disorder. PMID- 12773269 TI - Acute and transient psychotic disorders: comparison with schizophrenia. AB - Concept and nosologic status of acute and transient psychotic disorders, as they appear in the tenth edition of the International Classification of Disease, have seen review from the standpoint of validation and delineation from schizophrenia and affective disorders. Current research, particularly on the epidemiology, course, and outcome, and family genetic studies indicate that these disorders are common among women in developing countries, as well as among lower socioeconomic status and rural subjects. These patients have greater frequency of exposure to stress before childbirth, a family history of acute and transient psychotic disorder (and not of schizophrenia), and a course and outcome that is different from that of schizophrenia. The findings so far support the argument that acute and transient psychotic disorders are different from schizophrenia. PMID- 12773272 TI - Psychotic disorder and traumatic brain injury. AB - Traumatic brain injury (TBI) can result in serious and disabling neuropsychiatric disorders, such as cognitive deficits and personality change, as well as severe and chronic psychosis. This review focuses on the relationship between TBI and schizophrenia-like psychosis (SLP) including its epidemiology, diagnostic criteria, clinical presentation, psychopathology, risk factors, and pathophysiology. The relationships between post-traumatic epilepsy and SLP, and brain trauma and schizophrenia, are also discussed. The risk of SLP does increase after TBI. The clinical presentation has considerable overlap with primary schizophrenic disorder, with a prominence of persecutory and other delusions and auditory hallucinations, as well as a lack of negative symptoms. The onset is often gradual, with a subacute or chronic course. More severe and diffuse brain injury, especially of the temporal and frontal lobes, is the most prominent risk factor. Genetic load may also play a role, but presence of epilepsy could be a protective factor. Further large and systematic longitudinal studies are needed. PMID- 12773273 TI - Schizoaffective disorder: clinical aspects, differential diagnosis, and treatment. AB - Schizoaffective disorders are a clinical reality and their concept has a long history, but there are still problems regarding their diagnostic definition and nosologic categorization. The present definitions of the International Classification of Disease and the Diagnostic and Statistical Manual of Mental Disorders are not sufficient enough to define schizoaffective disorders, especially because of deficits on the longitudinal axis. Schizoaffective disorders occupy a position between schizophrenia and pure mood disorders, especially regarding prognosis and premorbid and sociodemographic variables. Schizoaffective disorders are recurrent disorders and, therefore, need prophylactic treatment. Suicidal symptomatology is extremely frequent in patients with schizodepressive episodes. The most severe type of schizoaffective disorders is the schizoaffective mixed type. Schizoaffective patients with mixed episodes retire more frequently and at younger ages than other patients with bipolar disorder. PMID- 12773271 TI - Outcome of schizophreniform disorder. AB - The aim of this review is to find data about the outcome of schizophreniform disorder. As different definitions of schizophreniform disorder were found in the literature, it was not surprising that data on its outcome, and on the relationship of schizophreniform disorder to schizophrenia and to mood disorders (as this relationship is linked to outcome), were often different and opposite. Its classic description of an acute onset psychotic episode with mood instability and a relatively brief duration should be the focus of future studies. Current studies, apart from a small number, lump together different (and probably distinct) subtypes of schizophreniform disorder (one with good prognostic features, and one without good prognostic features, according to the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition). Studies of mixed samples led to different results, probably depending on the relative prevalence of one subtype over the other one. The few studies on schizophreniform disorder with good prognostic features found more often an episodic, recurrent course, and a family history of mood disorders. These features link this schizophreniform disorder subtype more to mood disorders than to schizophrenia. If confirmed by future studies, these preliminary findings can have very important treatment implications, given the very different treatment strategies in mood disorders compared with schizophrenia. PMID- 12773274 TI - Gender differences in antidepressant response. PMID- 12773275 TI - Fibromyalgia: an overview. AB - In this article, the authors review current concepts in fibromyalgia. Findings regarding diagnosis, prevalence, comorbidities, and potential pathophysiologic links are discussed. Although fibromyalgia continues to be a complex disorder, there are specific criteria one must meet. Fibromyalgia questionnaires, along with commonalities of age, gender, menopause status, sleep disturbances, and mood symptoms, may aid in the diagnosis. Additionally, the close relationship between fibromyalgia and other chronic disorders should alert the physician to explore for comorbid illness. The relationship between fibromyalgia and irritable bowel syndrome, migraine headaches, and obesity are addressed. The roles of the hypothalamic-pituitary-axis, potential effects of neurotransmitters, and gender specific hormones all substantiate this diagnosis and provide clues to causality, as well as venues for future treatment. PMID- 12773276 TI - Management of fibromyalgia. AB - Fibromyalgia is characterized by widespread pain, persistent fatigue, nonrestorative sleep, and generalized morning stiffness. The diagnosis is based on patients' reports of pain and fatigue, clinical findings of multiple tender points, and exclusion of a range of connective tissue and other medical disorders. Treatment of fibromyalgia is multidisciplinary with an emphasis on active patient participation, medications, cognitive behavioral therapy, and physical modalities. No single medication has been found to effectively control all the symptoms, and a rational combination of different medications is often necessary. Currently available medication classes include the selective serotonin uptake inhibitors, the serotonin and norepinephrine reuptake inhibitors, tricyclic antidepressants, analgesics, hypnotic agents, and anticonvulsants. Treatment modalities should be individualized for patients based on target symptoms and impairment in functioning. As is the case with several chronic disorders, the treatment is often prolonged and improvement may occur slowly. Patience and positive attitude on part of the physician and active involvement of patients and their families in treatment are likely to enhance improvement. PMID- 12773278 TI - Cerebrovascular diseases and depression. AB - Cerebrovascular diseases constitute a leading health hazard. The association between stroke and depression has been recognized for many years. Depression is the most common psychiatric disorder associated with cerebrovascular diseases, most episodes of post-stroke depression occur in the first 2 years after a cerebrovascular accident. Studies have found an association between lesion location, physical impairment, cognitive impairment, aphasia, and post-stroke depression. The location of the lesion in terms of proximity to the left frontal pole of the brain has a profound impact on the frequency and severity of post stroke depression. Treatment modalities include pharmacotherapy, psychotherapy, electroconvulsive therapy, and rehabilitation. Understanding the psychologic and physical morbidity of post-stroke depression, as well as its timely, comprehensive treatment, are important for effective management. PMID- 12773281 TI - Intake of calcium-rich foods, body image, and self-esteem. PMID- 12773277 TI - Electroconvulsive therapy in the medically ill. AB - The use of electroconvulsive therapy (ECT) in the US continues to expand in the treatment of psychiatric disorders. Electroconvulsive therapy remains the "gold standard" for the treatment of major depression and a variety of other psychiatric and neurologic disorders. Because of the effectiveness and resurgence of ECT, more patients are considered good candidates for this treatment option. Overall, these patients are medication refractory and elderly, and thus more sensitive to polypharmacy. Additionally, these patients tend to have more coexisting medical problems, and often require that a practitioner have solid clinical knowledge of the fields of medicine and psychiatry. This article will review some of the most common comorbid conditions found in patients referred for ECT. PMID- 12773282 TI - Influences on diet and physical activity among middle-class African American 8- to 10-year-old girls at risk of becoming obese. AB - OBJECTIVE: To understand diet, physical activity, and inactivity influences among preadolescent African American girls at risk of becoming obese. DESIGN: Interviews and group qualitative discussions (i.e., mixed qualitative research method) were conducted separately with 8- to 10-year-old African American girls and their parents. SETTING: Greater Houston, Texas. PARTICIPANTS: Eight- to 10 year-old African American girls above the 50th percentile body mass index with a home computer (n = 82) and a parent (n = 74). VARIABLES MEASURED: Influences on dietary practices and physical activity/inactivity among preadolescent African American girls. ANALYSIS: Discussions were audiotaped, manually recorded, transcribed, and coded. The primary coder analyzed the transcribed notes. The secondary coder reviewed and critiqued the initial coding. RESULTS: Parents and girls were concerned about overweight and viewed physical activity as a weight control practice. Mothers facilitated daughters' physical activity, while fathers and siblings were coparticipants. Girls had access to physical activity equipment and facilities. Snack food items and carbonated beverages were often limited by the parents, and water consumption was encouraged. Discrepancies were apparent between girls' and parents' responses. CONCLUSIONS AND IMPLICATIONS: Parental convenience and girls' food preferences influenced dietary intake. Obesity prevention programs should capitalize on parental motivation for their child's health and provide practical strategies to facilitate healthful eating and physical activity. PMID- 12773284 TI - Women's perspectives on nutrition, health, and breast cancer. AB - OBJECTIVE: To explore women's beliefs about diet, health, and breast cancer. DESIGN: Individual interviews exploring women's perceptions of their eating habits, health status, and diet, health, and breast cancer beliefs. SETTING: Vancouver, British Columbia, Canada. PARTICIPANTS: A culturally diverse convenience sample of women aged 40 to 60 years, including breast cancer survivors (n = 29) and women who had not had breast cancer (n = 32). ANALYSIS: Verbatim interview transcripts were coded and themes developed by sorting and summarizing coded transcript segments. RESULTS: Three perspectives on healthful eating were identified. A "traditional" perspective stressed regular meals of meat, potatoes, and vegetables and made no specific claims about the diet-breast cancer relationship. A "mainstream" perspective stressed increasing vegetables and fruits and decreasing fat intake to improve health, reduce cardiovascular disease risk, and, possibly, reduce breast cancer risk. An "alternative" perspective focused on the role of toxins, carcinogens, and protective factors in food in affecting cancer risk. CONCLUSIONS AND IMPLICATIONS: Although overall perspectives on healthful eating were not related to participants' breast cancer status, they were related to how participants thought about the relationships between diet and breast cancer. Specific diet-cancer beliefs, degree of conviction about those beliefs, and reasons why foods are believed to be protective or harmful are indicative of overall nutrition perspectives. PMID- 12773283 TI - Accuracy of fourth-graders' dietary recalls of school breakfast and school lunch validated with observations: in-person versus telephone interviews. AB - OBJECTIVE: To investigate the accuracy of children's dietary recalls of school breakfast and school lunch validated with observations and obtained during in person versus telephone interviews. DESIGN: Each child was observed eating school breakfast and school lunch and was interviewed that evening about that day's intake. SETTING: Ten elementary schools. PARTICIPANTS: A sample of fourth-graders was randomly selected within race (black, white) and gender strata, observed, and interviewed in person (n = 33) or by telephone (n = 36). MAIN OUTCOMES MEASURED: Rates for omissions (items observed but not reported) and intrusions (items reported but not observed) were calculated to determine accuracy for reporting items. A measure of total inaccuracy was calculated to determine inaccuracy for reporting items and amounts combined. ANALYSIS: Analysis of variance; chi-square. RESULTS: Interview type (in person, telephone) did not significantly affect recall accuracy. For omission rate, intrusion rate, and total inaccuracy, means were 34%, 19%, and 4.6 servings for in-person recalls and 32%, 16%, and 4.3 servings for telephone recalls of school breakfast and school lunch. CONCLUSIONS AND IMPLICATIONS: The accuracy of children's recalls of school breakfast and school lunch is not significantly different whether obtained in person or by telephone. Whether interviewed in person or by telephone, children reported only 67% of items observed; furthermore, 17% of items reported were not observed. PMID- 12773285 TI - Mothers' acculturation and eating behaviors of Korean American families in California. AB - OBJECTIVE: To compare eating behaviors of Korean American (KA) families with mothers of different acculturation stages with those of families in Korea. DESIGN: Survey research using a self-report questionnaire. SETTINGS: Field survey on convenience sample in California and in Seoul, Korea. PARTICIPANTS: 225 KA and 216 Korean mothers. MAIN OUTCOME MEASURES: Frequency of eating at home, preparation of Korean foods, favorite dishes of families, and for special occasions. ANALYSIS: Chi-square, t test, and analysis of variance. RESULTS: Families of the more acculturated mothers dined out more frequently and had lower preferences for Korean foods than did the families with less acculturated mothers. The more acculturated mothers prepared Korean foods, including kimchi, less at home. Fewer Korean dishes were included among the 5 favored dishes, both for family and for special occasions, in KA families than in Korean families. The acculturation stage of the mothers affected preference for Korean dishes of the families but not for special occasions. CONCLUSIONS AND IMPLICATIONS: Mothers' acculturation stage influenced the family's eating habits. Changes of preference of typical Korean dishes varied by acculturation stage. Nutrition educators should be aware that dietary behaviors of KA families may differ by acculturation stage and tailor interventions appropriately. PMID- 12773286 TI - Food insecurity and food supplies in Latino households with young children. AB - OBJECTIVE: To examine the relationship between food insecurity and food supplies in Latino households. DESIGN: Cross-sectional survey, conducted February to May 2001. SETTING: Six California counties. PARTICIPANTS: Convenience sampling was used to recruit 274 low-income Latino families with preschool children from the Special Supplemental Nutrition Program for Women, Infants and Children (WIC), Head Start, and other community-based organizations. Complete data were available for 256 families. VARIABLES MEASURED: Food security, household food scores. ANALYSIS: Pearson correlations, Kruskal-Wallis test, and logistics regression. Significance level at P <.05. RESULTS: Controlling for maternal education, food insecurity over the past 3 months was associated with lower household food supplies: dairy, r = -.18, P <.01; fruit, r = -.36, P <.001; grains, r = -.27, P <.0001; meats, r = -.22, P <.001; snack foods, r = -.23, P <.001; and vegetables, r = -.29, P <.001. CONCLUSIONS AND IMPLICATIONS: In Latino households, greater food insecurity is associated with a lower variety of most foods, particularly fruits and vegetables. Future research in Latino households should explore the effects of seasonal food insecurity and household food shortages on food intake of individual household members, especially young children. PMID- 12773287 TI - Use of the think aloud method to examine fruit and vegetable purchasing behaviors among low-income African American women. AB - The purpose of this report is to describe the development and implementation of the think aloud method in relation to fruit and vegetable purchasing behaviors of low-income African American mothers. Women (n = 70) were audiotaped as they thought aloud while selecting fruits and vegetables during a routine shopping trip. Audiotapes were transcribed, text was coded, and coded text was sorted using a database software program. Data were analyzed using content analysis procedures. The method was found to be useful in its ability to provide verbalization data for the majority of the women in the sample that reflected a typical shopping experience, were not excessively affected by the presence of the investigator, and captured information processing in relation to salient factors that influenced food purchasing decisions. Because a few women indicated that the method itself may have influenced behavior, future research is needed to test the reactivity of the think aloud method and its relationship to final choice of products. PMID- 12773288 TI - Integrating service-learning into a college-level nutrition course. PMID- 12773291 TI - MR imaging of epilepsy: state of the art at 1.5 T and potential of 3 T. AB - Shortly after being introduced in the nineteen eighties, magnetic resonance imaging (MRI) became a key tool for the investigation of patients with epilepsy, due to its ability to acquire high quality images. The strength of the magnetic field of a scanner is measured in tesla (T). This review addresses the clinical and research potential in epilepsy of MR imaging at 1.5 T and 3 T. A typical clinical scanning protocol at 1.5 T for a patient with refractory epilepsy may include T1- and T2-weighted imaging, fluid-attenuated inversion recovery (FLAIR) imaging, and a 3D volume acquisition sequence. A research protocol may add quantification of structural imaging, such as volumetric assessment and T2 relaxometry, together with functional measures, such as MR-spectroscopy, functional MRI and diffusion weighted sequences. MR-spectroscopy assesses the metabolites of the seizure focus and other brain areas. Functional MRI allows localisation of cognitive and sensori-motor function and the ability to assess the spatial relationship of these functions to the seizure focus. Whereas these techniques can be performed at 1.5 T, particularly MR-spectroscopy and functional MRI benefit from increased magnetic field-strength. Higher magnetic field strength is associated with a higher signal-to-noise ratio (SNR). The increased SNR can allow shorter imaging times for a given resolution, higher resolution for a given imaging time, or combination of both. The use of higher magnetic field strengths is therefore indicated for the (fast) imaging of ill subjects, for long protocols, including structural, metabolic and functional imaging, and for novel applications, such as continuous EEG recording and functional MRI for the detection of the seizure focus. Disadvantages of MR imaging in epilepsy at a high field-strength of 3 T and above are, apart from engineering and technical challenges, the greater energy deposition into tissue and increased susceptibility to artefacts. So far, magnets of 3 T and above have been used mainly for research applications, however the benefits of high field-strength for MR spectroscopy and functional MRI, and the usefulness of these techniques for the investigation of epilepsy patients are obvious incentives for the use of 3 T systems in routine clinical investigations. PMID- 12773293 TI - Epilepsy and hypothalamic hamartoma: look at the hand Pallister-Hall syndrome. AB - We report the case of a 29-year-old patient, who suffered from drug resistant laughing seizures since childhood. The clinical examination was normal, except for sequelae of hand and feet surgery during infancy for post-axial polydactyly. Cerebral MRI showed a hypothalamic hamartoma. The association of complex limb abnormalities with hypothalamic hamartoma lead to the diagnosis of Pallister-Hall syndrome. This syndrome is related to a mutation of gene GLI3, located on chromosome 7p13, and its inheritance is autosomal dominant. In the case of laughing seizures, a cerebral MRI should be performed to look for a hypothalamic hamartoma. The observation of such lesions indicates the necessity of standard radiographies of the hands and feet, to search for associated abnormalities. These findings might help to recognize a Pallister-Hall syndrome, thus allowing genetic counseling. PMID- 12773292 TI - Familial severe myoclonic epilepsy of infancy: truncation of Nav1.1 and genetic heterogeneity. AB - BACKGROUND: Severe myoclonic epilepsy of infancy (SMEI) or Dravet syndrome has been long suspected of having a genetic origin. Recently, mutations in SCN1A and GABRG2 have been described in SMEI patients. The sporadic nature of the SMEI syndrome and the occurrence of SCN1A and GABRG2 mutations in a mild familial phenotype, termed generalized epilepsy with febrile seizure plus complicates genotype-phenotype correlations. In order to further investigate the role of SCN1A and GABRG2 in the pathogenesis of SMEI we have screened for mutations three families with at least two members affected by Dravet syndrome. METHODS: Clinical criteria followed the international classification of epileptic syndromes. Mutational screening of SCN1A and GABRG2 genes was performed by denaturing high performance liquid chromatography (DHPLC) and direct sequencing of DNA fragments showing a variant chromatogram. RESULTS: Thirty-eight fragments spanning 26 exons of SCN1A and nine exons of GABRG2 were analysed in three probands. Five variant chromatograms were identified; four corresponded to known polymorphisms, one to a novel dinucleotide insertion on exon 26 of SCN1A. The mutation leads to a frameshift and a premature stop codon at amino acid 1 779 of the protein. The mutation was present in the affected sibling and was inherited from the mother who had experienced a single febrile seizure in childhood. CONCLUSIONS: Among three families analysed, a single family was mutant for SCN1A. Our study suggests that the syndrome is genetically heterogeneous. The variable expressivity we observed for the c5240insAA mutation suggests that other factors are needed for the development of the full SMEI phenotype. PMID- 12773294 TI - Continuous giggling and autistic disorder associated with hypothalamic hamartoma. AB - We present the case of a child affected since early infancy from recurring episodes of giggling mixed with stereotypical behaviours, mingled with head drops, and eventually with falls, in the context of an autistic disorder. Scalp video-EEG recordings revealed an epileptic encephalopathy with generalized slow spike-and-wave complexes alternating with electrodecremental periods, which generally corresponded to the onset of the aforementioned clinical sequences. A resection of a hypothalamic hamartoma was achieved at the age of two. Since then, after 22 months of follow-up, the child is totally free from the giggling and the drops, and the autistic behaviour significantly improved during the second year of follow-up. This case illustrates the difficulties to recognize some subtle ictal manifestations during infancy and childhood, and encourages the early surgical treatment of hypothalamic hamartomas when associated with epileptic encephalopathy and when technically possible. PMID- 12773295 TI - Mesial temporal lobe epilepsy with focal photoparoxysmal response. AB - Intermittent photic stimulation (IPS) may produce epileptiform discharges and seizures, most of which are generalized. There are several cases of focal seizures of occipital origin induced by IPS and only five reported cases originating from the temporal lobe (TL). We report an unusual case of TL epilepsy, supported by electroclinical and neuroimaging data, with rhythmic focal TL discharges precipitated by photic stimulus. CASE REPORT: A 37 year old male with refractory epilepsy due to a right mesial temporal sclerosis presented a focal right TL photoparoxysmal response during IPS. A routine, interictal SPECT study obtained with injection of HMPAO, carried out during continuous pulses of the photic stimulus at 16 Hz, disclosed hyperperfusion over the right TL. Presurgical ictal studies demonstrated seizures arising from the same region. A temporal lobectomy rendered the patient seizure-free and further EEGs with IPS were normal. CONCLUSION: To our knowledge, this is the sixth case in medical literature showing evidence of photosensitivity associated with a TL lesion and it is the first in which photic stimulation induced a focal electrographic response of the involved TL. We hypothesize that in rare cases, photosensitivity may be related to a mesial TL lesion without occipital cortex involvement. PMID- 12773296 TI - Benign familial and non-familial infantile seizures: a study of 64 patients. AB - INTRODUCTION: The recently proposed diagnostic scheme for people with epileptic seizures and with epilepsy (Epilepsia 2001) includes two idiopathic focal epileptic syndromes with onset during the first year of life, the benign familial and non-familial infantile seizures. OBJECTIVES: To analyze the electroclinical features and evolution in patients with benign familial and non-familial infantile seizures. PATIENTS AND METHODS: Sixty-four patients (36 males and 28 females) were evaluated at the Neurology Department of the J. P. Garrahan Children's Hospital between February 1990 and December 2001. We analyzed gender, age at onset, duration, manifestations, circadian distribution and frequency of seizures, family history of epilepsy and paroxysmal dyskinesias. EEG and neuroradiological studies were performed. The semeiology of the seizures was analyzed only according to the description in the clinical history. Ictal EEGs could not be recorded. RESULTS: Twenty-five patients, 14 girls and 11 boys, had a family history of similar seizures with an age at seizure onset of 3 to 22 months (median of 5.5 months). Nine patients (36%) had apparently generalized convulsions only; five patients (20%) partial seizures only and ten patients (50%) had both partial and generalized seizures. Convulsions were brief, during wakefulness in all, and occurred in clusters in 12 patients (48%). Interictal EEG was normal in 24 patients (96%). Similar seizures and age at onset were found in 14 fathers, ten mothers and one uncle of these patients. Twenty-two patients (88%) had their last seizure before the age of 30 months. Two siblings of the same family later had brief episodes of paroxysmal kinesigenic dyskinesia, which in one of them were associated with infantile seizures. Later, two fathers developed paroxysmal kinesigenic dystonia. One of them also had had infantile seizures. A second group of 39 patients (25 boys and 14 girls) showed similar electroclinical features and evolution but there was no history of infantile seizures in first-degree relatives family history of epilepsy was found in 12.8% of second-degree relatives, but the type of epilepsy could not be defined. CONCLUSION: This study confirms the existence of a familial benign epileptic syndrome in infancy, of probable dominant autosomical transmission. A large group also had similar electroclinical features but without a family history. We discuss the possible relationships between the two groups and suggest that further genetic studies may solve the problem. PMID- 12773297 TI - Pre-clinical studies with the GABAergic compounds vigabatrin and tiagabine. AB - In the last decade, nine new antiepileptic drugs have reached the global marketplace. These new agents can be categorised according to their principal mechanisms of action. Vigabatrin and tiagabine are the only new compounds with selective effects on inhibitory neurotransmission. The principal inhibitory neurotransmitter in mammalian brain is gamma-aminobutyric acid (GABA). Both vigabatrin and tiagabine exert their pharmacological effects by reducing the inactivation of GABA. Vigabatrin attenuates the metabolism of GABA by inhibiting the enzyme GABA-transaminase, whereas tiagabine blocks the uptake of GABA from the synaptic cleft by an action on the GAT-1 transporter. These mechanistic differences are borne out in a range of experimental seizure models in which vigabatrin and tiagabine have very different anticonvulsant profiles. Pre clinical neurotoxicity and pharmacokinetic profiles also differ. Long-term vigabatrin treatment is associated with intramyelinic oedema in white matter tracts of several brain regions and further studies have revealed an accumulation of vigabatrin in the retina. In contrast, it seems that tiagabine does not precipitate any significant neurotoxicity and does not appear to accumulate in the retina. The results of these pre-clinical investigations suggest that vigabatrin and tiagabine are pharmacologically distinct compounds with different anticonvulsant, neurotoxicity and pharmacokinetic profiles. It is possible that they will ultimately prove to have different clinical efficacies and spectra of activity. PMID- 12773298 TI - Benign idiopathic occipital epilepsy: report of a case of the late (Gastaut) type [corrected]. AB - We report the electro-clinical characteristics of a case with Gastaut type idiopathic occipital epilepsy. The visual seizures, a cardinal feature of the syndrome, are described and illustrated by video sequences. A number of stereotyped seizures are shown where the child, an 11-year-old boy, clearly describes the visual phenomena experienced. Prognosis of this common form of partial idiopathic epilepsy is relatively good but, as in our case, seizures may be very frequent during a long period of time. In such cases the term 'benign' is probably not the most appropriate. PMID- 12773299 TI - Prolonged infusion of angiotensin II into normal rats induces stellate cell activation and proinflammatory events in liver. AB - Recent evidence indicates that angiotensin II (ANG II) plays an important role in liver fibrogenesis. However, the underlying mechanisms are largely unknown. In advanced chronic liver diseases, circulating levels of ANG II are frequently elevated. We investigated the hepatic effects of prolonged systemic infusion of ANG II in normal rats. Saline or ANG II at subpressor and pressor doses (15 and 50 ng.kg-1.min-1, respectively) were infused to normal rats for 4 wk through a subcutaneous osmotic pump. Infusion of ANG II resulted in liver injury, as assessed by elevated serum liver enzymes. Livers from ANG II-perfused rats showed activation of JNK and ERK as well as increased NF-kappaB and activating protein-1 DNA-binding activity. Moreover, ANG II perfusion induced oxidative stress, increased concentration of proinflammatory cytokines, and upregulated the inflammatory proteins inducible nitric oxide synthase and cyclooxygenase-2. Histological examination of the livers from ANG II-infused rats showed mild portal inflammation as well as thickening and thrombosis of small hepatic vessels. ANG II-treated livers showed accumulation of CD43-positive inflammatory cells and activated hepatic stellate cells (HSCs) at the pericentral areas. A slight increase in collagen synthesis was observed, as assessed by Sirius red staining and hepatic hydroxyproline. All of these effects were observed when ANG II was perfused at subpressor and pressor doses. ANG II also accelerated the activation of primary cultured rat HSCs. In conclusion, increased systemic ANG II can induce liver injury by promoting proinflammatory events and vascular damage. ANG II-induced hepatic effects are not dependent on increase in arterial pressure. PMID- 12773300 TI - Cultured gallbladder epithelial cells synthesize apolipoproteins A-I and E. AB - Gallbladder epithelial cells (GBEC) are exposed to high and fluctuating concentrations of biliary cholesterol on their apical (AP) surface. GBEC absorb and efflux cholesterol, but the mechanisms of cholesterol uptake, intracellular trafficking, and efflux in these cells are not known. We previously reported that ATP binding cassette (ABC)A1 mediates basolateral (BL) cholesterol efflux in cultured polarized GBEC. In addition, the nuclear hormone receptors liver X receptor (LXR)alpha and retinoid X receptor (RXR) mediate both AP and BL cholesterol efflux. An interesting finding from our previous study was that apolipoprotein (apo)A-I applied to the AP surfaces of cells elicited BL ABCA1 mediated cholesterol efflux. Because ABCA1-mediated cholesterol efflux requires the presence of a cholesterol acceptor, we hypothesized that GBEC synthesize and secrete endogenous apo into the BL compartment. Here, we demonstrate that cholesterol loading of cells with model bile and AP apoA-I treatment is associated with an increase in the synthesis of apoE mRNA and protein. Furthermore, apoE is secreted into the BL compartment. LXRalpha/RXR ligands stimulate the synthesis of endogenous apoA-I mRNA and protein, as well as apoE mRNA. BL secretion of apoA-I is elicited by LXRalpha/RXR ligands. Therefore, GBEC synthesize apoA-I and -E and efflux cholesterol using ABCA1- and non-ABCA1- mediated pathways. These processes may alter gallbladder biliary cholesterol concentrations and thereby influence gallstone formation. PMID- 12773301 TI - IL-6 is essential for development of gut barrier dysfunction after hemorrhagic shock and resuscitation in mice. AB - We sought to determine the role of IL-6 as a mediator of the alterations in gut barrier function that occur after hemorrhagic shock and resuscitation (HS/R). C57Bl/6 wild-type (WT) and IL-6 knockout (KO) mice on a C57Bl/6 background were subjected to either a sham procedure or HS/R. Organ and tissue samples were obtained 4 h after resuscitation. In WT mice, HS/R significantly increased ileal mucosal permeability to fluorescein isothiocyanate-labeled dextran (average molecular mass, 4 kDa) and bacterial translocation to mesenteric lymph nodes. These alterations in gut barrier function were not observed in IL-6 KO animals. HS/R increased ileal steady-state mRNA levels for IL-6, TNF, and IL-10 in WT but not in IL-6 KO mice. Ileal mucosal expression of the tight junction protein, ZO 1, decreased after HS/R in WT but not IL-6 KO mice. Collectively, these data support the view that expression of IL-6 is essential for the development of gut barrier dysfunction after HS/R. PMID- 12773302 TI - Caerulein-induced acute pancreatitis inhibits protein synthesis through effects on eIF2B and eIF4F. AB - Acute pancreatitis (AP) has been shown in some studies to inhibit total protein synthesis in the pancreas, whereas in other studies, protein synthesis was not affected. Previous in vitro work has shown that high concentrations of cholecystokinin both inhibit protein synthesis and inhibit the activity of the guanine nucleotide exchange factor eukaryotic initiation factor (eIF)2B by increasing the phosphorylation of eIF2alpha. We therefore evaluated in C57BL/6 mice the effects of caerulein-induced AP on pancreatic protein synthesis, eIF2B activity and other protein translation regulatory mechanisms. Repetitive hourly injections of caerulein were administered at 50 microg/kg ip. Pancreatic protein synthesis was reduced 10 min after the initial caerulein administration and was further inhibited after three and five hourly injections. Caerulein inhibited the two major regulatory points of translation initiation: the activity of the guanine nucleotide exchange factor eIF2B (with an increase of eIF2alpha phosphorylation) and the formation of the eIF4F complex due, in part, to degradation of eIF4G. This inhibition was not accounted for by changes in the upstream stimulatory pathway, because caerulein activated Akt as well as phosphorylating the downstream effectors of mTOR, 4E-BP1, and ribosomal protein S6. Caerulein also decreased the phosphorylation of the eukaryotic elongation factor 2, implying that this translation factor was not inhibited in AP. Thus the inhibition of pancreatic protein synthesis in this model of AP most likely results from the inhibition of translation initiation as a result of increased eIF2alpha phosphorylation, reduction of eIF2B activity, and the inhibition of eIF4F complex formation. PMID- 12773303 TI - Suppression of glucose production by GLP-1 independent of islet hormones: a novel extrapancreatic effect. AB - Glucagon-like peptide-1 (GLP-1) is an intestinal hormone that stimulates insulin secretion and decreases glucagon release. It has been hypothesized that GLP-1 also reduces glycemia independent of its effect on islet hormones. Based on preliminary evidence that GLP-1 has independent actions on endogenous glucose production, we undertook a series of experiments that were optimized to address this question. The effect of GLP-1 on glucose appearance (Ra) and glucose disposal (Rd) was measured in eight men during a pancreatic clamp that was performed by infusing octreotide to suppress secretion of islet hormones, while insulin and glucagon were infused at rates adjusted to maintain blood glucose near fasting levels. After stabilization of plasma glucose and equilibration of [3H]glucose tracer, GLP-1 was given intravenously for 60 min. Concentrations of insulin, C-peptide, and glucagon were similar before and during the GLP-1 infusion (115 +/- 14 vs. 113 +/- 11 pM; 0.153 +/- 0.029 vs. 0.156 +/- 0.026 nM; and 64.7 +/- 11.5 vs. 65.8 +/- 13.8 ng/l, respectively). With the initiation of GLP-1, plasma glucose decreased in all eight subjects from steady-state levels of 4.8 +/- 0.2 to a nadir of 4.1 +/- 0.2 mM. This decrease in plasma glucose was accounted for by a significant 17% decrease in Ra, from 22.6 +/- 2.8 to 19.1 +/- 2.8 micromol. kg-1. min-1 (P < 0.04), with no significant change in Rd. These findings indicate that, under fasting conditions, GLP-1 decreases endogenous glucose production independent of its actions on islet hormone secretion. PMID- 12773304 TI - Insulin effects on acetate metabolism. AB - Acetate metabolism was studied in patients with insulin resistance. To evaluate the interaction between glucose and acetate metabolism, we measured acetate and glucose turnover with a hyperinsulinemic euglycemic clamp (hot clamp) in obese and diabetic patients with insulin resistance (n = 8) and in a control group with normal insulin sensitivity (n = 6). At baseline, acetate turnover and plasma concentrations were similar between the two groups (group means: 4.3 +/- 0.4 micromol x kg-1 x min-1 and 128.2 +/- 11.1 micromol/l). Acetate concentrations decreased in both groups with hyperinsulinemia but were significantly lower in the insulin-resistant group (20% vs. 12%, P < 0.05). After the hot clamp treatment, acetate turnover increased for the two groups and was higher in the group with normal insulin sensitivity: 8.1 +/- 0.7 vs. 5.5 +/- 0.5 micromol x kg 1 x min-1 (P < 0.001). No change related to insulin action was observed in either group in the percentage of acetate oxidation. This was approximately 70% of overall utilization at baseline and during the clamp. No correlation between glucose and acetate utilization was observed. Our results support the hypothesis that, like glucose metabolism, acetate metabolism is sensitive to insulin. PMID- 12773305 TI - Characterization of rat iodothyronine sulfotransferases. AB - Sulfation appears to be an important pathway for the reversible inactivation of thyroid hormone during fetal development. The rat is an often used animal model to study the regulation of fetal thyroid hormone status. The present study was done to determine which sulfotransferases (SULTs) are important for iodothyronine sulfation in the rat, using radioactive T4, T3, rT3, and 3,3'-T2 as substrates, 3'-phosphoadenosine-5'-phosphosulfate (PAPS) as cofactor, and rat liver, kidney and brain cytosol, and recombinant rat SULT1A1, -1B1, -1C1, -1E1, -2A1, -2A2, and -2A3 as enzymes. Recombinant rat SULT1A1, -1E1, -2A1, -2A2, and -2A3 failed to catalyze iodothyronine sulfation. For all tissue SULTs and for rSULT1B1 and rSULT1C1, 3,3'-T2 was by far the preferred substrate. Apparent Km values for 3,3' T2 amounted to 1.9 microM in male liver, 4.4 microM in female liver, 0.76 microM in male kidney, 0.23 microM in male brain, 7.7 microM for SULT1B1, and 0.62 microM for SULT1C1, whereas apparent Km values for PAPS showed less variation (2.0-6.9 microM). Sulfation of 3,3'-T2 was inhibited dose dependently by other iodothyronines, with similar structure-activity relationships for most enzymes except for the SULT activity in rat brain. The apparent Km values of 3,3'-T2 in liver cytosol were between those determined for SULT1B1 and -1C1, supporting the importance of these enzymes for the sulfation of iodothyronines in rat liver, with a greater contribution of SULT1C1 in male than in female rat liver. The results further suggest that rSULT1C1 also contributes to iodothyronine sulfation in rat kidney, whereas other, yet-unidentified forms appear more important for the sulfation of thyroid hormone in rat brain. PMID- 12773306 TI - Chronic hepatic artery ligation does not prevent liver from differentiating portal vs. peripheral glucose delivery. AB - Infusion of glucose into the hepatic artery blocks the stimulatory effect of the "portal signal" on net hepatic glucose uptake (NHGU) during portal glucose delivery. We hypothesized that hepatic artery ligation (HAL) would result in enhanced NHGU during peripheral glucose infusion because the arterial glucose concentration would be perceived as lower than that in the portal vein. Fourteen dogs underwent HAL approximately 16 days before study. Conscious 42-h-fasted dogs received somatostatin, intraportal insulin, and glucagon infusions at fourfold basal and at basal rates, respectively, and peripheral glucose infusion to create hyperglycemia. After 90 min (period 1), seven dogs (HALpo) received intraportal glucose (3.8 mg. kg-1. min-1) and seven (HALpe) continued to receive only peripheral glucose for 90 min (period 2). These two groups were compared with nine non-HAL control dogs (control) treated as were HALpe. During period 2, the arterial plasma insulin concentrations (24 +/- 3, 20 +/- 1, and 24 +/- 2 microU/ml) and hepatic glucose loads (39.1 +/- 2.5, 43.8 +/- 2.9, and 37.7 +/- 3.7 mg. kg-1. min-1) were not different in HALpe, HALpo, and control, respectively. HALpo exhibited greater (P < 0.05) NHGU than HALpe and control (3.1 +/- 0.3, 2.0 +/- 0.4, and 2.0 +/- 0.1 mg. kg-1. min-1, respectively). Net hepatic carbon retention was approximately twofold greater (P < 0.05) in HALpo than in HALpe and control. NHGU and net hepatic glycogen synthesis during peripheral glucose infusion were not enhanced by HAL. Even though there exists an intrahepatic arterial reference site for the portal vein glucose concentration, the failure of HAL to result in enhanced NHGU during peripheral glucose infusion suggests the existence of one or more comparison sites outside the liver. PMID- 12773307 TI - Endothelin-1-stimulated glucose uptake is desensitized by tumor necrosis factor alpha in 3T3-L1 adipocytes. AB - Tumor necrosis factor-alpha (TNF-alpha) is a potent inducer of insulin resistance, and increased TNF-alpha expression is associated with impaired glucose disposal. Although insulin is the primary regulator of glucose transport in adipose, endothelin-1, a vasoconstrictor peptide that signals through the heterotrimeric G proteins Galphaq/11, potently stimulates glucose uptake in 3T3 L1 adipocytes by a mechanism independent of phosphatidylinositol (PI) 3-kinase. Here, we report that exposure of 3T3-L1 adipocytes to TNF-alpha for 48 h dose dependently decreased endothelin-1-stimulated glucose uptake and translocation of GLUT4 to the plasma membrane. TNF-alpha exposure had no effect on endothelin-1 receptor number at the cell surface. In contrast, TNF-alpha treatment reduced the quantity of Galphaq/11 and proline-rich tyrosine kinase 2 (PYK2) and decreased endothelin-1-stimulated PYK2-Tyr402 tyrosine phosphorylation. Taken together, these results suggest that TNF-alpha-induced desensitization of endothelin-1 stimulated GLUT4 translocation and glucose uptake in 3T3-L1 adipocytes is due, at least in part, to a decreased expression of Galphaq/11, leading to a suppression in tyrosine phosphorylation of PYK2. PMID- 12773308 TI - Endotoxin induces differential regulation of mTOR-dependent signaling in skeletal muscle and liver of neonatal pigs. AB - In the present study, differential responses of regulatory proteins involved in translation initiation in skeletal muscle and liver during sepsis were studied in neonatal pigs treated with lipopolysaccharide (LPS). LPS did not alter eukaryotic initiation factor (eIF) 2B activity in either tissue. In contrast, binding of eIF4G to eIF4E to form the active mRNA-binding complex was repressed in muscle and enhanced in liver. Phosphorylation of eIF4E-binding protein, 4E-BP1, and ribosomal protein S6 kinase, S6K1, was reduced in muscle during sepsis but increased in liver. Finally, changes in 4E-BP1 and S6K1 phosphorylation were associated with altered phosphorylation of the protein kinase mammalian target of rapamycin (mTOR). Overall, the results suggest that translation initiation in both skeletal muscle and liver is altered during neonatal sepsis by modulation of the mRNA-binding step through changes in mTOR activation. Moreover, the LPS induced changes in factors that regulate translation initiation are more profound than previously reported changes in global rates of protein synthesis in the neonate. This finding suggests that the initiator methionyl-tRNA-rather than the mRNA-binding step in translation initiation may play a more critical role in maintaining protein synthesis rates in the neonate during sepsis. PMID- 12773309 TI - Obesity in transgenic female mice with constitutively elevated luteinizing hormone secretion. AB - Transgenic (TG) female mice, expressing a chimeric bovine luteinizing hormone (LH) beta-subunit/human chorionic gonadotropin beta-subunit COOH-terminal extension (bLHbeta-CTP) gene, produce high levels of circulating LH and serve as a model for functional ovarian hyperandrogenism and follicular cysts. We report here that obesity is a typical feature of these female mice. The mean body weight of the bLHbeta-CTP females was significantly higher than in controls at, and beyond 5 wk of age, and at 5 mo, it was 32% increased. At this age, the amount of white adipose tissue in the bLHbeta-CTP females was significantly increased, as reflected by the weight difference of the retroperitoneal fat pad. In addition, the expression of leptin mRNA in white adipose tissue of the TG females was elevated about twofold. Serum leptin and insulin levels, and food intake, were also increased significantly in the TG females. Brown adipose tissue (BAT) thermogenic activity, as measured by GDP binding to BAT mitochondria, was reduced (P < 0.05). Ovariectomy at the age of 3 wk totally prevented the development of obesity. In summary, the present results show that intact female bLHbeta-CTP mice are obese, have increased food consumption, and reduced BAT thermogenic activity. The weight gain can be explained partly by elevated androgens but is probably also contributed to the increased adrenal steroidogenesis. Hence, the bLHbeta-CTP mice provide a useful model for studying obesity related to elevated LH secretion, with consequent alterations in ovarian and adrenal function. PMID- 12773310 TI - Hydrogen peroxide stimulates ubiquitin-conjugating activity and expression of genes for specific E2 and E3 proteins in skeletal muscle myotubes. AB - Reactive oxygen species (ROS) are thought to promote muscle atrophy in chronic wasting diseases, but the underlying mechanism has not been determined. Here we show that H2O2 stimulates ubiquitin conjugation to muscle proteins through transcriptional regulation of the enzymes (E2 and E3 proteins) that conjugate ubiquitin to muscle proteins. Incubation of C2C12 myotubes with 100 microM H2O2 increased the rate of 125I-labeled ubiquitin conjugation to muscle proteins in whole cell extracts. This response required at least 4-h exposure to H2O2 and persisted for at least 24 h. Preincubating myotubes with cycloheximide or actinomycin D blocked H2O2 stimulation of ubiquitin-conjugating activity, suggesting that gene transcription is required. Northern blot analyses revealed that H2O2 upregulates expression of specific E3 and E2 proteins that are thought to regulate muscle catabolism, including atrogin1/MAFbx, MuRF1, and E214k. These results suggest that ROS stimulate protein catabolism in skeletal muscle by upregulating the ubiquitin conjugation system. PMID- 12773311 TI - Lanthanum is transported by the sodium/calcium exchanger and regulates its activity. AB - La3+ uptake was measured in fura 2-loaded Chinese hamster ovary cells expressing the bovine cardiac Na+/Ca2+ exchanger (NCX1.1). La3+ was taken up by the cells after an initial lag phase of 50-60 s and achieved a steady state within 5-6 min. Neonatal cardiac myocytes accumulated La3+ in a similar manner. La3+ uptake was due to the activity of the exchanger, because no uptake was seen in nontransfected cells or in transfected cells that had been treated with gramicidin to remove cytosolic Na+. The low rate of La3+ uptake during the lag period resulted from insufficient cytosolic Ca2+ to activate the exchanger at its regulatory sites, as shown by the following observations. La3+ uptake occurred without a lag period in cells expressing a mutant of NCX1.1 that does not exhibit regulatory activation by cytosolic Ca2+. The rate of La3+ uptake by wild-type cells was increased, and the lag phase was reduced or eliminated, when the cytosolic Ca2+ concentration was increased before initiating La3+ uptake. La3+ could substitute for Ca2+ at very low concentrations to activate exchange activity. Thus preloading cells expressing NCX1.1 with a small quantity of La3+ increased the rate of exchange-mediated Ca2+ influx by 20-fold; in contrast, cytosolic La3+ partially inhibited Ca2+ uptake by the regulation-deficient mutant. With an estimated KD of 30 pM for the binding of La3+ to fura 2, we conclude that cytosolic La3+ activates exchange activity at picomolar concentrations. We speculatively suggest that endogenous trace metals might activate exchange activity under physiological conditions. PMID- 12773312 TI - Expression of SERCA2a is independent of innervation in regenerating soleus muscle. AB - The speed of contraction of a skeletal muscle largely depends on the myosin heavy chain isoforms (MyHC), whereas the relaxation is initiated and maintained by the sarcoplasmic reticulum Ca2+-ATPases (SERCA). The expression of the slow muscle type myosin heavy chain I (MyHCI) is entirely dependent on innervation, but, as we show here, innervation is not required for the expression of the slow-type sarcoplasmic reticulum Ca2+-ATPase (SERCA2a) in regenerating soleus muscles of the rat, although it can play a modulator role. Remarkably, the SERCA2a level is even higher in denervated than in innervated regenerating soleus muscles on day 7 when innervation is expected to resume. Later, the level of SERCA2a protein declines in denervated regenerated muscles but it remains expressed, whereas the corresponding mRNA level is still increasing. SERCA1 (i.e., the fast muscle-type isoform) expression shows only minor changes in denervated regenerating soleus muscles compared with innervated regenerating controls. When the soleus nerve was transected instead of the sciatic nerve, SERCA2a and MyHCI expressions were found to be even more uncoupled because the MyHCI nearly completely disappeared, whereas the SERCA2a mRNA and protein levels decreased much less. The transfection of regenerating muscles with constitutively active mutants of the Ras oncogene, known to mimic the effect of innervation on the expression of MyHCI, did not affect SERCA2a expression. These results demonstrate that the regulation of SERCA2a expression is clearly distinct from that of the slow myosin in the regenerating soleus muscle and that SERCA2a expression is modulated by neuronal activity but is not entirely dependent on it. PMID- 12773313 TI - Cross-bridge blocker BTS permits direct measurement of SR Ca2+ pump ATP utilization in toadfish swimbladder muscle fibers. AB - Because the major processes involved in muscle contraction require rapid utilization of ATP, measurement of ATP utilization can provide important insights into the mechanisms of contraction. It is necessary, however, to differentiate between the contribution made by cross-bridges and that of the sarcoplasmic reticulum (SR) Ca2+ pumps. Specific and potent SR Ca2+ pump blockers have been used in skinned fibers to permit direct measurement of cross-bridge ATP utilization. Up to now, there was no analogous cross-bridge blocker. Recently, N benzyl-p-toluene sulfonamide (BTS) was found to suppress force generation at micromolar concentrations. We tested whether BTS could be used to block cross bridge ATP utilization, thereby permitting direct measurement of SR Ca2+ pump ATP utilization in saponin-skinned fibers. At 25 microM, BTS virtually eliminates force and cross-bridge ATP utilization (both <4% of control value). By taking advantage of the toadfish swimbladder muscle's unique right shift in its force Ca2+ concentration ([Ca2+]) relationship, we measured SR Ca2+ pump ATP utilization in the presence and absence of BTS. At 25 microM, BTS had no effect on SR pump ATP utilization. Hence, we used BTS to make some of the first direct measurements of ATP utilization of intact SR over a physiological range of [Ca2+]at 15 degrees C. Curve fits to SR Ca2+ pump ATP utilization vs. pCa indicate that they have much lower Hill coefficients (1.49) than that describing cross-bridge force generation vs. pCa (approximately 5). Furthermore, we found that BTS also effectively eliminates force generation in bundles of intact swimbladder muscle, suggesting that it will be an important tool for studying integrated SR function during normal motor behavior. PMID- 12773314 TI - More than apical: Distribution of SGLT1 in Caco-2 cells. AB - We investigated the distribution of the endogenous sodium-d-glucose cotransporter (SGLT1) in polarized Caco-2 cells, a model for enterocytes. A cellular organelle fraction was separated by free-flow electrophoresis and subjected to the analysis of endogenous and exogenous marker enzymes for various membrane vesicle components. Furthermore, the presence of SGLT1 was tested by an ELISA assay employing newly developed epitope specific antibodies. Thereby it was found that the major amount of SGLT1 resided in intracellular compartments and only a minor amount in apical plasma membranes. The distribution ratio between intracellular SGLT1 and apical membrane-associated SGLT1 was approximately 2:1. Further immunocytochemical investigation of SGLT1 distribution in fixed Caco-2 cells by epifluorescence and confocal microscopy revealed that the intracellular compartments containing SGLT1 were associated with microtubules. Elimination of SGLT1 synthesis by incubation of cells with cycloheximide did not significantly reduce the size of the intracellular SGLT1 pool. Furthermore, the half-life of SGLT1 in Caco-2 cells was determined to be 2.5 days by metabolic labeling followed by immunoprecipitation. Our data suggest that most of the intracellular SGLT1 are not transporters en route from biosynthesis to their cellular destination but represent an intracellular reserve pool. We therefore propose that intracellular compartments containing SGLT1 are involved in the regulation of SGLT1 abundance at the apical cell surface. PMID- 12773315 TI - Transcriptional regulation of IGF-I expression in skeletal muscle. AB - The present study investigated the role of transcription in the regulation of insulin-like growth factor (IGF)-I expression in skeletal muscle. RT-PCR was used to determine endogenous expression of IGF-I pre-mRNA and mRNA in control (Con) and functionally overloaded (FO) rat plantaris. The transcriptional activities of five different-length IGF-I promoter fragments controlling transcription of a firefly luciferase (FLuc) reporter gene were tested in vitro by transfection of myoblasts or in vivo during FO by direct gene transfer into the plantaris. Increased endogenous IGF-I gene transcription during 7 days of plantaris FO was evidenced by an approximately 140-160% increase (P < 0.0001) in IGF-I pre-mRNA (a transcriptional marker). IGF-I mRNA expression also increased by approximately 90% (P < 0.0001), and it was correlated (R = 0.93; P < 0.0001) with the pre-mRNA increases. The three longest IGF-I exon 1 promoters induced reporter gene expression in proliferating C2C12 and L6E9 myoblasts. In differentiated L6E9 myotubes, promoter activity increased approximately two- to threefold over myoblasts. Overexpression of calcineurin and MyoD increased the activity of the 852/+192 promoter in C2C12 myotubes by approximately 5- and approximately 18 fold, respectively. However, FO did not induce these exogenous promoter fragments. Nevertheless, the present findings are consistent with the hypothesis that the IGF-I gene is transcriptionally regulated during muscle hypertrophy in vivo as evidenced by the induction of the endogenous IGF-I pre-mRNA during plantaris FO. The exon 1 promoter region of the IGF-I gene is sufficient to direct inducible expression in vitro; however, an in vivo response to FO may require elements outside the -852/+346 region of the exon 1 IGF-I promoter or features inherent to the endogenous IGF-I gene. PMID- 12773316 TI - Plasma membrane delivery of the gastric H,K-ATPase: the role of beta-subunit glycosylation. AB - The factors determining trafficking of the gastric H,K-ATPase to the apical membrane remain elusive. To identify such determinants in the gastric H,K-ATPase, fusion proteins of yellow fluorescent protein (YFP) and the gastric H,K-ATPase beta-subunit (YFP-beta) and cyan fluorescent protein (CFP) and the gastric H,K ATPase alpha-subunit (CFP-alpha) were expressed in HEK-293 cells. Then plasma membrane delivery of wild-type CFP-alpha, wild-type YFP-beta, and YFP-beta mutants lacking one or two of the seven beta-subunit glycosylation sites was determined using confocal microscopy and surface biotinylation. Expression of the wild-type YFP-beta resulted in the plasma membrane localization of the protein, whereas the expressed CFP-alpha was retained intracellularly. When coexpressed, both CFP-alpha and YFP-beta were delivered to the plasma membrane. Removing each of the seven glycosylation sites, except the second one, from the extracellular loop of YFP-beta prevented plasma membrane delivery of the protein. Only the mutant lacking the second glycosylation site (Asn103Gln) was localized both intracellularly and on the plasma membrane. A double mutant lacking the first (Asn99Gln) and the second (Asn103Gln) glycosylation sites displayed intracellular accumulation of the protein. Therefore, six of the seven glycosylation sites in the beta-subunit are essential for the plasma membrane delivery of the beta subunit of the gastric H,K-ATPase, whereas the second glycosylation site (Asn103), which is not conserved among the beta-subunits from different species, is not critical for plasma delivery of the protein. PMID- 12773317 TI - Effects of osmotic swelling on voltage-gated calcium channel currents in rat anterior pituitary cells. AB - Decrease in extracellular osmolarity ([Os]e) results in stimulation of hormone secretion from pituitary cells. Different mechanisms can account for this stimulation of hormone secretion. In this study we examined the possibility that hyposmolarity directly modulates voltage-gated calcium influx in pituitary cells. The effects of hyposmolarity on L-type (IL) and T-type (IT) calcium currents in pituitary cells were investigated by using two hyposmotic stimuli, moderate (18 22% decrease in [Os]e) and strong (31-32% decrease in [Os]e). Exposure to moderate hyposmotic stimuli resulted in three response types in IL (a decrease, a biphasic effect, and an increase in IL) and in increase in IT. Exposure to strong hyposmotic stimuli resulted only in increases in both IL and IT. Similarly, in intact pituitary cells (perforated patch method), exposure to either moderate or strong hyposmotic stimuli resulted only in increases in both IL and IT. Thus it appears that the main effect of decrease in [Os]e is increase in calcium channel currents. This increase was differential (IL were more sensitive than IT) and voltage independent. In addition, we show that these hyposmotic effects cannot be explained by activation of an anionic conductance or by an increase in cell membrane surface area. In conclusion, this study shows that hyposmotic swelling of pituitary cells can directly modulate voltage-gated calcium influx. This hyposmotic modulation of IL and IT may contribute to the previously reported hyposmotic stimulation of hormone secretion. The mechanisms underlying these hyposmotic effects and their possible physiological relevance are discussed. PMID- 12773318 TI - Our strategies for fighting severe acute respiratory syndrome (SARS). PMID- 12773319 TI - The role of innate immunity in acute allograft rejection after lung transplantation. AB - Although innate immunity is crucial to pulmonary host defense and can initiate immune and inflammatory responses independent of adaptive immunity, it remains unstudied in the context of transplant rejection. To investigate the role of innate immunity in the development of allograft rejection, we assessed the impact of two functional polymorphisms in the toll-like receptor 4 (TLR4) associated with endotoxin hyporesponsiveness on the development of acute rejection after human lung transplantation. Patients and donors were screened for the TLR4 Asp299Gly and Thr399Ile polymorphisms by polymerase chain reaction using sequence specific primers. The rate of acute rejection at 6 months was significantly reduced in recipients, but not in donors, with the Asp299Gly or Thr399Ile alleles as compared with wild type (29 vs. 56%, respectively, p = 0.05). This association was confirmed in Cox proportional hazards and multivariate logistic regression models. Our results suggest activation of innate immunity in lung transplant recipients through TLR4 contributes to the development acute rejection after lung transplantation. Therapies directed at inhibition of innate immune responses mediated by TLR4 may represent a novel and effective means to prevent acute rejection after lung transplantation. PMID- 12773320 TI - Complement receptor 1 gene polymorphisms are associated with idiopathic pulmonary fibrosis. AB - Idiopathic pulmonary fibrosis (IPF) is a chronic, fibrotic disorder underlain by aberrant wound healing of repeated lung injury. Environmental triggers and genetic background are likely to act as modifiers of the fibrotic response. Erythrocyte complement receptor 1 is a membrane protein mediating the transport of immune complexes to phagocytes. Three gene polymorphisms are related to the erythrocyte surface density of complement receptor 1 molecules, which in turn are related to the rate of immune complexes' clearance. There is evidence of association between sarcoidosis and the complement receptor 1 gene. We wondered whether IPF is associated with the complement receptor 1 gene alleles coding for a reduced molecule/erythrocyte ratio. We studied 74 patients and 166 control subjects. Three polymorphic sites of the gene, A3650G exon 22, HindIII RFLP intron 27, and C5507G exon 33, were analyzed and found to be in linkage disequilibrium. The GG genotype for the C5507G exon 33 polymorphism was significantly more common in patients with IPF than in control subjects (odds ratio = 6.232, 95% confidence interval = 2.198-18.419, p = 0.00023). The significant difference was found in both sexes. These findings agree with speculations on the role of the complement receptor 1 gene in idiopathic pulmonary fibrosis. PMID- 12773321 TI - Contributions of upper airway mechanics and control mechanisms to severity of obstructive apnea. AB - The contributions of pharyngeal mechanical abnormalities, flow demand, and compensatory effectiveness to obstructive sleep apnea severity were determined in 82 patients. Flow demand was estimated from mean inspiratory flow on continuous positive airway pressure. Mechanical load on upper airway muscles was estimated from minimal effective continuous positive airway pressure, flow demand, and minimum flow observed during brief pressure dial downs. Compensatory effectiveness was estimated by relating polysomnographic severity and mechanical load. Mechanical load was more severe in men, in supine position, and in older and heavier patients. Higher flow demand contributed significantly to mechanical load in men and in those who are obese. At the same mechanical load, severity was independent of age, sex, or body mass index but was greater in the supine position and in REM sleep. Mechanical load accounted for only 34% of variability in severity. Eighty-two percent of patients experienced periods of stable breathing despite mechanical loads that would produce continuous cycling without compensation. I conclude that most patients can adequately compensate for the abnormal mechanics, at least part of the time. Higher flow demand contributes to severity in men and in obesity. Severity is largely due to factors other than mechanical load. Compensatory effectiveness is impaired in the supine position and in REM sleep, but not by age, sex, or body mass index. PMID- 12773322 TI - Risk factors for tuberculosis infection in sub-Saharan Africa: a contact study in The Gambia. AB - Few studies have investigated the risk factors for tuberculosis (TB) infection in highly endemic countries. We conducted a household study in The Gambia, in which a tuberculin skin test (TST) was performed in members of the households of 315 smear-positive pulmonary TB cases and 305 community control subjects. The risk of being TST positive (10 mm or more) was higher in contacts of cases than in contacts of control subjects. It increased with age, male sex, and duration of stay in the household but was not associated with the presence of a bacille de Calmette-Guerin scar. Within the households of the TB cases, the risk of TST positivity was higher in males and was increased with age, social proximity to the case, and the radiologic extent of the disease in the case's chest X-ray. Adjusting on these, the risk of TST positivity was higher in first-degree relatives compared with more distant relatives and nongenetically related household members, but the effect was not statistically significant. In highly endemic areas, the risk of TB infection in contacts of TB infectious cases is associated with age, sex, intensity of exposure to the case, and severity of disease in the case, but it is possible that genetic factors contribute to the susceptibility to Mycobacterium tuberculosis infection. PMID- 12773323 TI - Cellular antiendotoxin activities of lung surfactant protein C in lipid vesicles. AB - The respiratory system is continuously exposed to airborne particles containing lipopolysaccharide. Our laboratory established previously that the hydrophobic surfactant protein C (SP-C) binds to lipopolysaccharide and to one of its cellular receptors, CD14. Here we examined the influence of SP-C, and of a synthetic analog, on some cellular in vitro effects of lipopolysaccharide. When associated with vesicles of dipalmitoylphosphatidylcholine, SP-C inhibits the binding of a tritium-labeled lipopolysaccharide to the macrophage cell line RAW 264.7. Under similar conditions of presentation, SP-C inhibits the mitogenic effect of lipopolysaccharide on mouse splenocytes, and inhibits the lipopolysaccharide-induced production of tumor necrosis factor-alpha by peritoneal and alveolar macrophages, and of nitric oxide by RAW 264.7 cells. In contrast, tumor necrosis factor-alpha production induced by a lipopeptide, and nitric oxide production induced by picolinic acid, were not affected by SP-C. The lipopolysaccharide-binding capacity of SP-C is resistant to peroxynitrite, a known mediator of acute lung injury formed by reaction of nitric oxide with superoxide anions. These results indicate that SP-C may play a role in lung defense; SP-C resists degradation under inflammatory conditions and traps lipopolysaccharide, preventing it from inducing production of noxious mediators in alveolar cells. PMID- 12773324 TI - Snoring, intermittent hypoxia and academic performance in primary school children. AB - Sleep-disordered breathing is associated with impaired attention and neurocognitive deficits. We assessed the association of snoring and intermittent hypoxia with poor academic performance in third grade school children (1,144 children). Snoring frequency and intermittent hypoxia were investigated using parental questionnaire and nocturnal home pulse oximetry. Intermittent hypoxia was specified as desaturation events of 90% or less pulse oximeter saturation. Poor academic performance was defined as grade 4-6 on a six-point scale (i.e., approximately the lowest quintile grades) in mathematics, science, reading, spelling, and/or handwriting in the most recent school report. Snoring "always" was significantly associated with poor academic performance in mathematics (odds ratio; 95% confidence interval: 3.6; 1.3-10.1), science (4.3; 1.3-14.6), and spelling (3.5; 1.2-10.3). Snoring "frequently" was also significantly associated with poor academic performance in mathematics (2.4; 1.3-4.7) and spelling (2.0; 1.04-3.8). A significant relationship between snoring and poor academic performance was also found in children without intermittent hypoxia, whereas intermittent hypoxia did not show an independent association with poor academic performance. Thus, habitual snoring (i.e., snoring frequently or always) was associated with poor academic performance in these primary school children. PMID- 12773325 TI - Changes in clinical and physiologic variables predict survival in idiopathic pulmonary fibrosis. AB - There is significant heterogeneity in survival time among patients with idiopathic pulmonary fibrosis. Studies of baseline clinical and physiologic variables as predictors of survival time have reported inconsistent results. We evaluated the predictive value of changes in clinical and physiologic variables over time for survival time in 81 patients with biopsy-proven idiopathic pulmonary fibrosis. Six-month changes in dyspnea score, total lung capacity, thoracic gas volume, FVC, FEV1, diffusing capacity of carbon monoxide, partial pressure of arterial oxygen, oxygen saturation, and alveolar-arterial oxygen gradient were predictive of survival time even after adjustment for baseline values. Analyses were repeated on 51 patients with 12-month change data. Twelve month changes in dyspnea score, total lung capacity, FVC, partial pressure of arterial oxygen, oxygen saturation, and alveolar-arterial oxygen gradient were predictive of survival time after adjustment for baseline values. Evaluation of changes in clinical and physiological variables over 6 and 12 months may provide clinicians with more accurate prognostic information than baseline values alone. PMID- 12773326 TI - Cyclooxygenase 2 and intermittent hypoxia-induced spatial deficits in the rat. AB - Intermittent hypoxia (IH) during sleep, a critical feature of sleep apnea, induces significant neurobehavioral deficits in the rat. Cyclooxygenase (COX)-2 is induced during stressful conditions such as cerebral ischemia and could play an important role in IH-induced learning deficits. We therefore examined COX-1 and COX-2 genes and COX-2 protein expression and activity (prostaglandin E2 [PGE2] tissue concentration) in cortical regions of rat brain after exposure to either IH (10% O2 alternating with 21% O2 every 90 seconds) or sustained hypoxia (10% O2). In addition, the effect of selective COX-2 inhibition with NS-398 on IH induced neurobehavioral deficits was assessed. IH was associated with increased COX-2 protein and gene expression from Day 1 to Day 14 of exposure. No changes were found in COX-1 gene expression after exposure to hypoxia. IH-induced COX-2 upregulation was associated with increased PGE2 tissue levels, neuronal apoptosis, and neurobehavioral deficits. Administration of NS-398 abolished IH induced apoptosis and PGE2 increases without modifying COX-2 mRNA expression. Furthermore, NS-398 treatment attenuated IH-induced deficits in the acquisition and retention of a spatial task in the water maze. We conclude that IH induces upregulation and activation of COX-2 in rat cortex and that COX-2 may play a role in IH-mediated neurobehavioral deficits. PMID- 12773327 TI - Serotonin transporter inhibitors protect against hypoxic pulmonary hypertension. AB - Pulmonary hypertension (PH) results from constriction and remodeling of pulmonary vessels. Serotonin contributes to both phenomena through different signaling pathways. The mitogenic effect of serotonin on pulmonary vascular smooth muscle cells is mediated by the serotonin transporter (5-hydroxytryptamine transporter [5-HTT]), whereas its constricting effect is mediated by 5-HT1B/1D and 5-HT2A receptors. Here, we investigated the respective roles of 5-HTT and 5-HT receptors on the development of chronic hypoxic PH in mice. During exposure to hypoxia (10% O2 for 2 weeks), the animals received one of the specific 5-HTT inhibitors citalopram and fluoxetine (10 mg/kg/day), the selective 5-HT1B/1D receptor antagonist GR127935 (2 and 10 mg/kg/day), or the 5-HT2A receptor antagonist ketanserin (2 mg/kg/day). Mice treated with the 5-HTT inhibitors showed less right ventricle hypertrophy (ratio of right ventricle/left ventricle + septum = 36.7 +/- 2.0% and 35.8 +/- 1.3% in citalopram- and fluoxetine-treated mice, respectively, vs. 41.5 +/- 1.5% in vehicle-treated mice) and less pulmonary vessel muscularization (p < 0.01) than those receiving the vehicle. Neither GR127935 nor ketanserin affected these parameters. These data indicate that 5-HTT plays a key role in hypoxia-induced pulmonary vascular remodeling. The effects of serotonin transporter inhibitors on PH in humans deserve investigation. PMID- 12773328 TI - Type 1 and type 2 cytokine imbalance in acute respiratory syncytial virus bronchiolitis. AB - We examined the in vivo immune response of infants to natural respiratory syncytial virus (RSV) infection through analysis of cytokine levels in nasal lavage fluid and stimulated peripheral blood mononuclear cells. Eighty-eight babies with at least one parent with atopy and asthma were prospectively studied through their first winter. Twenty-eight infants had an upper respiratory tract infection where RSV was detected, of whom nine developed signs of acute bronchiolitis. Nasal lavage specimens were assayed for interferon-gamma, interleukin (IL)-4, IL-10, and IL-12 and the RSV load determined by quantitative polymerase chain reaction. Messenger RNA (mRNA) was extracted from stimulated peripheral blood mononuclear cells and interferon-gamma, IL-4, IL-12, and IL-18 mRNA levels determined by polymerase chain reaction. Cytokine profiles were analyzed in relation to clinical outcome. The IL-4/interferon-gamma ratio for infants with acute bronchiolitis was elevated in nasal lavage fluid on both Days 1-2 (p = 0.014) and Days 5-7 (p = 0.001) of the illness compared with infants with upper respiratory tract infection alone. Those with acute bronchiolitis demonstrated a higher IL-10/IL-12 ratio (p = 0.0015) on Days 1-2. IL-18 mRNA levels were reduced (p = 0.019) and the IL-4/interferon-gamma ratio elevated (p = 0.01) in stimulated peripheral blood mononuclear cells from infants with acute bronchiolitis. There was no difference in initial RSV load. These data strongly implicate excess type 2 and/or deficient type 1 immune responses in the pathogenesis of RSV bronchiolitis. PMID- 12773329 TI - Prognostic implications of physiologic and radiographic changes in idiopathic interstitial pneumonia. AB - Idiopathic interstitial pneumonias are a diverse group of lung diseases with varied prognoses. We hypothesized that changes in physiologic and radiographic parameters would predict survival. We retrospectively examined 80 patients with usual interstitial pneumonia and 29 patients with nonspecific interstitial pneumonia. Baseline characteristics were examined together with 6-month change in forced vital capacity, diffusing capacity for carbon monoxide, and ground glass infiltrate and fibrosis on high resolution computed tomography. Patients with usual interstitial pneumonia were more likely to have a statistically significant or marginally significant decline in lung volume, diffusing capacity for carbon monoxide, and an increase in ground glass infiltrates (p < or = 0.08) compared with patients with nonspecific interstitial pneumonia. For patients with usual interstitial pneumonia, change in forced vital capacity was the best physiologic predictor of mortality (p = 0.05). In a multivariate Cox proportional hazards model controlling for histopathologic diagnosis, gender, smoking history, baseline forced vital capacity, and 6-month change in forced vital capacity, a decrease in forced vital capacity remained an independent risk factor for mortality (decrease > 10%; hazard ratio 2.47; 95% confidence interval 1.29, 4.73; p = 0.006). We conclude that a 6-month change in forced vital capacity gives additional prognostic information to baseline features for patients with idiopathic interstitial pneumonia. PMID- 12773330 TI - Effects of acute hypoxia and lipopolysaccharide on nitric oxide synthase-2 expression in acute lung injury. AB - The potential role of nitric oxide synthase-2 (NOS2) in acute lung injury (ALI) has gained increasing attention. This study evaluates the effects of hypoxia, an important feature of ALI, on NOS2 expression in a rat model of ALI caused by exposure to hypoxia and LPS. Exposure to hypoxia alone had no effect on the expression of NOS2 in rat lungs. LPS treatment resulted in a significant increase in NOS2 in the lungs, which was further enhanced by concomitant exposure to hypoxia. Immunohistochemical analysis and in situ hybridization showed no changes in the expression of NOS2 in lung resident cells under any conditions. The increase in NOS2 levels is mainly due to the influx of NOS2-expressing inflammatory cells. By morphologic analysis, these inflammatory cells were identified as neutrophils, lymphocytes, and monocytes. In vitro experiments of lung epithelial and endothelial cell lines showed no detectable expression of NOS2 with any of the treatments. In a macrophage cell line, LPS-induced NOS2 expression was not affected by the concomitant exposure to hypoxia. In conclusion, LPS increases NOS2 expression in rat lungs through the recruitment of NOS2-producing leukocytes. Simultaneous exposure to LPS and hypoxia results in a greater influx of inflammatory cells that further enhances NOS2 expression. PMID- 12773331 TI - Sex differences in thoracic dimensions and configuration. AB - The volume of adult female lungs is typically 10-12% smaller than that of males who have the same height and age. In this study, we investigated how this volume difference is distributed between the rib cage and the diaphragm abdomen compartments. Internal rib cage dimensions, diaphragm position relative to spine, and diaphragm length were compared in 21 normal male and 19 normal female subjects at three different lung volumes using anterior-posterior and lateral chest radiographs. At all lung volumes examined, females had smaller radial rib cage dimensions in relationship to height than males, a greater inclination of ribs, a comparable diaphragm dome position relative to the spine, and a shorter diaphragm length. Female subjects exhibited a greater inspiratory rib cage muscle contribution during resting breathing than males, presumably reflecting an improved mechanical advantage conferred to these muscles by the greater inclination of ribs. Because of a greater inclination of ribs, female rib cages could accommodate a greater volume expansion. The results suggest a disproportionate growth of the rib cage in females relative to the lung, which would be well suited to accommodate large abdominal volume displacements as in pregnancy. PMID- 12773332 TI - Influence of talc dose on extrapleural talc dissemination after talc pleurodesis. AB - This study was designed to ascertain, in a rabbit model, extrapleural talc deposition and the related inflammatory response after talc slurry pleurodesis with two clinical doses, 200 and 50 mg/kg. Histopathologic evaluations revealed that whereas numerous rabbits receiving a high dose had talc in the ipsilateral (70%) and contralateral (55%) lung, mediastinum (90%), pericardium (30%), and liver (25%), a small number of animals treated with a low dose showed talc in the ipsilateral lung (10%) and mediastinum (20%) and none in the contralateral lung, pericardium, or liver. Hematologic and immunocytochemical analyses showed that a systemic inflammatory response develops shortly after pleurodesis with a high talc dose involving massive accumulation of neutrophils and macrophages in lung tissue. Zymography also revealed that the pulmonary expression of matrix metalloproteinases 2 and 9 was up-regulated in both lungs in a dose-dependent manner soon after talc instillation. Furthermore, microscopic examination of lung specimens revealed that the higher the dose of talc, the greater the development of both fibrotic visceral pleural thickening and foreign-body granulomas. These findings show pleurodesis with a high talc dose to be associated with an increased risk of extrapleural talc deposition, which may originate undesirable acute and chronic inflammatory responses. PMID- 12773333 TI - Transdiaphragmatic pressure control of airway pressure support in healthy subjects. AB - We designed a new servoventilator that proportionally adjusts airway pressure to transdiaphragmatic pressure (Pdi) generated by the subject during inspiration. Each cycle is triggered by either a preset Pdi increase or a preset inspiratory flow value (whichever is reached first), whereas cycling-off is flow-dependent. We evaluated the servoventilator in seven healthy subjects at normocapnia and three levels of hypercapnia (normocapnia + 3, + 6, and + 9 mm Hg) comparatively with spontaneous breathing. Triggering was by Pdi in six subjects and flow in one. At all end-tidal carbon dioxide pressure levels, time from onset of diaphragm electromyographic activity to inspiratory flow was similar with and without the servoventilator. Airway pressure increased proportionally to Pdi variation during servoventilator breathing. Flow, tidal volume, respiratory rate, intrinsic positive end-expiratory pressure, and esophageal and transdiaphragmatic pressure-time products increased significantly with hypercapnia with and without the servoventilator. Breathing pattern parameters were similar in the two breathing modes, and no differences were found for intrinsic positive end expiratory pressure or gastric pressure variation during exhalation. Esophageal and transdiaphragmatic pressure-time products were lower with than without the servoventilator. The Pdi-driven servoventilator was well synchronized to the subjects effort, delivering a pressure proportional to Pdi and reducing respiratory effort at normocapnia and hypercapnia. PMID- 12773335 TI - Odds ratios for suicide attempts among gay and bisexual men. PMID- 12773337 TI - The negative binomial process and arrests for drug use. PMID- 12773338 TI - HIV returns. PMID- 12773339 TI - When plagues don't end. PMID- 12773340 TI - Black men who have sex with men and the HIV epidemic: next steps for public health. PMID- 12773341 TI - Ruth Rice Puffer. PMID- 12773342 TI - Anthropological assessment for culturally appropriate interventions targeting men who have sex with men. AB - Although social and cultural factors play a fundamental role in the health of sexual minority populations and the development of culturally appropriate interventions, public health activities and research have sometimes lacked appropriate sophistication or attention to issues of cultural competency. In areas such as HIV prevention for men who have sex with men (MSM), biomedical interpretations of same-sex phenomena should be applied with caution. Communities and societies may broadly understand same-sex desire, attraction, behavior, and identity through age-structured/initiatory, gender-defined, profession/social role-defined, or egalitarian/gay frameworks. When more detailed, locally specific information is required, such as for youth, ethnic minorities, or urban versus rural populations, the approach to rapid anthropological assessment presented can provide nuanced insights for effective health programs targeting MSM. PMID- 12773344 TI - Field-delivered therapy increases treatment for chlamydia and gonorrhea. AB - In 1998, treatment completion was low among chlamydia and gonorrhea cases reported to the San Francisco Department of Public Health and assigned for treatment follow-up. To improve treatment completion among growing numbers of chlamydia and gonorrhea cases, the department implemented field-delivered therapy (FDT), a single-dose, directly observed therapy protocol for uncomplicated chlamydial and gonococcal infections. After the protocol was implemented in March 1999, the proportion of cases completing treatment increased significantly, from 61.8% in 1998 to 81.0% in 2000. The greatest increases in treatment completion were observed for females and individuals younger than 20 years old. FDT is an effective, feasible, and convenient way to reach and treat individuals who are unlikely to return for chlamydia and gonorrhea treatment. PMID- 12773343 TI - The second wave will drown us. PMID- 12773345 TI - Characterizing a "new" disease: epizootic and epidemic anthrax, 1769-1780. AB - In 1876, Robert Koch established anthrax as the first disease linked to a microbial agent. But Koch's efforts had followed more than 150 years of scientific progress in characterizing anthrax as a specific human and veterinary disease. Focusing on France and the period between 1769 and 1780, this brief review examines noteworthy early events in the characterization of anthrax. It suggests that some "new" diseases like anthrax might be "discovered" not only by luck, brilliance, or new technologies, but by clinical/epidemiological "puzzle fitting," which can assemble a cohesive picture of a seemingly specific disease entity. If such processes have operated over 2 or more centuries, studying them may yield clues about desirable interactions between epidemiology/public health and experimental science in the characterization of new diseases. PMID- 12773346 TI - Sexual behavior in the human male. 1948. PMID- 12773347 TI - Alfred C. Kinsey: a pioneer of sex research. PMID- 12773348 TI - One size does not fit all in the transgender community. PMID- 12773349 TI - Value of consistent condom use: a study of sexually transmitted disease prevention among African American adolescent females. PMID- 12773350 TI - HIV prevalence, risk behaviors, and high-risk sexual and injection networks among young women injectors who have sex with women. PMID- 12773351 TI - HIV and AIDS risk behaviors in juvenile detainees: implications for public health policy. PMID- 12773352 TI - Condom use and HIV risk among US adults. PMID- 12773353 TI - Validation of race/ethnicity and transmission mode in the US HIV/AIDS reporting system. PMID- 12773354 TI - HIV- and AIDS-related knowledge, awareness, and practices in Madagascar. PMID- 12773355 TI - Hepatitis B virus immunization among young injection drug users in San Francisco, Calif: the UFO Study. PMID- 12773356 TI - The use of complementary/alternative medicine therapies for the self-treatment of pain among residents of urban, suburban, and rural communities. PMID- 12773357 TI - High-risk behaviors among men who have sex with men in 6 US cities: baseline data from the EXPLORE Study. AB - OBJECTIVES: We describe the prevalence of risk behaviors at baseline among men who have sex with men (MSM) who were enrolled in a randomized behavioral intervention trial conducted in 6 US cities. METHODS: Data analyses involved MSM who were negative for HIV antibodies and who reported having engaged in anal sex with 1 or more partners in the previous year. RESULTS: Among 4295 men, 48.0% and 54.9%, respectively, reported unprotected receptive and insertive anal sex in the previous 6 months. Unprotected sex was significantly more likely with 1 primary partner or multiple partners than with 1 nonprimary partner. Drug and alcohol use were significantly associated with unprotected anal sex. CONCLUSIONS: Our findings support the continued need for effective intervention strategies for MSM that address relationship status, serostatus of partners, and drug and alcohol use. PMID- 12773358 TI - An individually tailored intervention for HIV prevention: baseline data from the EXPLORE Study. AB - OBJECTIVES: We describe the intervention tested in EXPLORE, an HIV prevention trial aimed at men who have sex with men (MSM), and test the empirical basis of the individually tailored intervention. METHODS: Data on participants' self efficacy, communication skills, social norms, and enjoyment of unprotected anal intercourse were examined in relation to sexual risk. Combinations of these factors, together with alcohol use and noninjection drug use, were also examined. RESULTS: The individual factors examined were associated with sexual risk behavior. The cohort was shown to be heterogeneous in regard to the presence of combinations of these risk-related factors. CONCLUSIONS: Baseline data from the EXPLORE study support the efficacy of the individually tailored intervention used. PMID- 12773359 TI - Association of co-occurring psychosocial health problems and increased vulnerability to HIV/AIDS among urban men who have sex with men. AB - OBJECTIVES: We measured the extent to which a set of psychosocial health problems have an additive effect on increasing HIV risk among men who have sex with men (MSM). METHODS: We conducted a cross-sectional household probability telephone sample of MSM in Chicago, Los Angeles, New York, and San Francisco. RESULTS: Psychosocial health problems are highly intercorrelated among urban MSM. Greater numbers of health problems are significantly and positively associated with high risk sexual behavior and HIV infection. CONCLUSIONS: AIDS prevention among MSM has overwhelmingly focused on sexual risk alone. Other health problems among MSM not only are important in their own right, but also may interact to increase HIV risk. HIV prevention might become more effective by addressing the broader health concerns of MSM while also focusing on sexual risks. PMID- 12773360 TI - HIV infections and associated costs attributable to syphilis coinfection among African Americans. AB - OBJECTIVES: We estimated the number and cost of syphilis-attributable HIV cases among African Americans. METHODS: A mathematical model of HIV transmission was used to estimate the number of partnerships consisting of HIV-discordant African Americans in which infectious syphilis was present and the number of new HIV cases attributable to syphilis in these partnerships. RESULTS: In 2000, an estimated 545 new cases of HIV infection among African Americans could be attributed to the facilitative effects of infectious syphilis, at a cost of about 113 million dollars. CONCLUSIONS: Syphilis prevention could reduce HIV incidence rates and the disproportionate burden of HIV/AIDS on the African American community, resulting in substantial reductions in future HIV/AIDS medical costs. PMID- 12773361 TI - Sex without disclosure of positive HIV serostatus in a US probability sample of persons receiving medical care for HIV infection. AB - OBJECTIVES: We estimated the proportion of HIV-positive adults who have any sexual contact without disclosure and the proportion of their sexual partnerships that involve unprotected sex without disclosure. METHODS: We drew participants from the HIV Cost and Services Utilization Study (n = 1421). Interviews assessed disclosure and sexual activities with up to 5 recent partners. RESULTS: Overall, 42% of the gay or bisexual men, 19% of the heterosexual men, and 17% of all the women reported any sex without disclosure, predominately within nonexclusive partnerships (P <.001). Across all groups, 13% of serodiscordant partnerships involved unprotected anal or vaginal sex without disclosure, with no significant difference between groups. CONCLUSIONS: Risky sex without disclosure of serostatus is not uncommon among people with HIV. PMID- 12773362 TI - Condom availability programs in Massachusetts high schools: relationships with condom use and sexual behavior. AB - OBJECTIVES: This study assessed relationships between condom availability programs accompanied by community discussion and involvement and adolescent sexual practices. METHODS: Sexual practice and condom use differences were assessed in a representative sample of 4166 adolescents enrolled in high schools with and without condom availability programs. RESULTS: Adolescents in schools where condoms were available were more likely to receive condom use instruction and less likely to report lifetime or recent sexual intercourse. Sexually active adolescents in those schools were twice as likely to use condoms, but less likely to use other contraceptive methods, during their most recent sexual encounter. CONCLUSIONS: The strategy of making condoms available, an indication of socioenvironmental support for condom use, may improve HIV prevention practices. PMID- 12773363 TI - The efficacy of a relationship-based HIV/STD prevention program for heterosexual couples. AB - OBJECTIVES: This study examined the efficacy of a relationship-based HIV/sexually transmitted disease prevention program for heterosexual couples and whether it is more effective when delivered to the couple or to the woman alone. METHODS: Couples (n = 217) were recruited and randomized to (1) 6 sessions provided to couples together (n = 81), (2) the same intervention provided to the woman alone (n = 73), or (3) a 1-session control condition provided to the woman alone (n = 63). RESULTS: The intervention was effective in reducing the proportion of unprotected and increasing the proportion of protected sexual acts. No significant differences in effects were observed between couples receiving the intervention together and those in which the woman received it alone. CONCLUSIONS: This study demonstrates the efficacy of a relationship-based prevention program for couples at risk for HIV infection. PMID- 12773364 TI - Rapid assessment of the HIV/AIDS crisis in racial and ethnic minority communities: an approach for timely community interventions. AB - OBJECTIVES: The US Department of Health and Human Services, in collaboration with the Congressional Black Caucus, created a new initiative to address the disproportionate ongoing HIV/AIDS crisis in racial/ethnic minority populations. METHODS: This initiative included deploying technical assistance teams through the Office of HIV/AIDS Policy. The teams introduced rapid assessment and response methodologies and trained minority communities in their use. RESULTS: The first 3 eligible cities (Detroit, Miami, and Philadelphia) focused assessments in small geographic areas, using multiple methodologies to obtain data. CONCLUSIONS: Data from the first 3 eligible cities provided critical information about changing the dynamics of the HIV/AIDS epidemic at the local level, including program and policy changes and infrastructure redeployment targeted at the most serious social and environmental conditions. PMID- 12773365 TI - Use of alternative therapists among people in care for HIV in the United States. AB - OBJECTIVES: This study examined the influence of sociodemographic, clinical, and attitudinal variables on the use of alternative therapists by people in care for HIV. METHODS: Bivariate and multivariate analyses of baseline data from the nationally representative HIV Cost and Services Utilization Study were conducted. RESULTS: Overall, 15.4% had used an alternative therapist, and among users, 53.9% had fewer than 5 visits in the past 6 months. Use was higher for people who were gay/lesbian, had incomes above 40,000 dollars, lived in the Northeast and West, were depressed, and wanted more information about and more decisionmaking involvement in their care. Among users, number of visits was associated with age, education, sexual orientation, insurance status, and CD4 count. CONCLUSIONS: Among people receiving medical care for HIV, use of complementary care provided by alternative therapists is associated with several sociodemographic, clinical, and attitudinal variables. Evaluation of the coordination of provider-based alternative and standard medical care is needed. PMID- 12773366 TI - The outing of Philip Morris: advertising tobacco to gay men. AB - OBJECTIVES: This case study describes the events surrounding the first time a major tobacco company advertised in gay media. METHODS: We analyzed internal tobacco company documents, mainstream newspapers, and the gay press. RESULTS: Philip Morris was unprepared for the attention its entry into the gay market received. The company's reaction to this incident demonstrates that its approach to the gay community both parallels and diverges from industry strategies toward other marginalized communities. CONCLUSIONS: The tobacco industry's relationship to the gay community is relatively undeveloped, a fact that may provide tobacco control advocates an opportunity for early intervention. The gay community's particular vulnerabilities to the industry make development of gay tobacco control programs crucial to reducing gay smoking prevalence and industry presence in the community. PMID- 12773367 TI - Cigarette smoking and cognitive decline in midlife: evidence from a prospective birth cohort study. AB - OBJECTIVES: The authors investigated the effects of cigarette smoking on midlife cognitive performance. METHODS: Multiple regression was used to test the association between cigarette smoking and changes in cognitive test scores among male and female members of the British 1946 birth cohort aged between 43 and 53 years. RESULTS: Smoking was associated with faster declines in verbal memory and with slower visual search speeds. These effects were largely accounted for by individuals who smoked more than 20 cigarettes per day and were independent of sex, socioeconomic status, previous (adolescent) cognitive ability, and a range of health indicators. CONCLUSIONS: The present results show that heavy smoking is associated with cognitive impairment and decline in midlife. Smokers who survive into later life may be at risk of clinically significant cognitive declines. PMID- 12773368 TI - Outcomes, safety, and resource utilization in a collaborative care birth center program compared with traditional physician-based perinatal care. AB - OBJECTIVE: We compared outcomes, safety, and resource utilization in a collaborative management birth center model of perinatal care versus traditional physician-based care. METHODS: We studied 2957 low-risk, low-income women: 1808 receiving collaborative care and 1149 receiving traditional care. RESULTS: Major antepartum (adjusted risk difference [RD] = -0.5%; 95% confidence interval [CI] = -2.5, 1.5), intrapartum (adjusted RD = 0.8%; 95% CI = -2.4, 4.0), and neonatal (adjusted RD = -1.8%; 95% CI = -3.8, 0.1) complications were similar, as were neonatal intensive care unit admissions (adjusted RD = -1.3%; 95% CI = -3.8, 1.1). Collaborative care had a greater number of normal spontaneous vaginal deliveries (adjusted RD = 14.9%; 95% CI = 11.5, 18.3) and less use of epidural anesthesia (adjusted RD = -35.7%; 95% CI = -39.5, -31.8). CONCLUSIONS: For low risk women, both scenarios result in safe outcomes for mothers and babies. However, fewer operative deliveries and medical resources were used in collaborative care. PMID- 12773369 TI - Rethinking the socioeconomics and geography of tuberculosis among foreign-born residents of New Jersey, 1994-1999. AB - OBJECTIVES: This study investigated the socioeconomic profile of foreign-born tuberculosis patients in New Jersey. METHODS: Foreign- and US-born tuberculosis patients in 1994-1999 were compared using various measures of socioeconomic status. RESULTS: Out of 4295 tuberculosis patients, 2005 (47%) were foreign-born. Foreign-born patients resided in more affluent, more educated, and less crowded areas than did US-born patients (P <.005). They were also more likely to have been employed during the 2 years before diagnosis (62% vs 41%, P <.001). Private physicians treated the majority of South Asian-born patients. CONCLUSIONS: Substantial numbers of employed foreign-born tuberculosis patients now reside in affluent New Jersey locations. Changes in tuberculosis control programs may be required when the socioeconomic status and place of residence of foreign-born populations diverge from traditional assumptions linking poverty with tuberculosis. PMID- 12773370 TI - Regulation of neuronal survival and death by extracellular signals during development. AB - Cell death is a prominent feature of the developing vertebrate nervous system, affecting neurons, glial cells and their progenitors. The most extensively studied and best understood phase of cell death occurs in populations of neurons shortly after they begin establishing connections with other neurons and/or non neural tissues. This phase of cell death makes appropriate adjustments to the relative sizes of interconnected groups of neurons and matches the size of neuronal populations that innervate non-neural tissues to the optimal requirements of these tissues. The fate of neurons during this period of development is regulated by a variety of secreted proteins that either promote survival or bring about cell death after binding to receptors expressed on the neurons. These proteins may be derived from the targets the neurons innervate, the afferents they receive or from associated glial cells, or they may be secreted by the neurons themselves. In this review, I will outline the established and emerging principles that modulate neuronal number in the developing nervous system. PMID- 12773371 TI - Dendritic cell regulation of immune responses: a new role for interleukin 2 at the intersection of innate and adaptive immunity. AB - Dendritic cells are professional antigen-presenting cells able to initiate innate and adaptive immune responses against invading pathogens. In response to external stimuli dendritic cells undergo a complete genetic reprogramming that allows them to become, soon after activation, natural killer cell activators and subsequently T cell stimulators. The recent observation that dendritic cells produce interleukin 2 following microbial stimulation opens new possibilities for understanding the efficiency of dendritic cells in regulating immune system functions. This review discusses how dendritic cells control natural killer, T- and B-cell responses and the relevance of interleukin 2 in these processes. PMID- 12773372 TI - NF-kappaB and virus infection: who controls whom. AB - Among the different definitions of viruses, 'pirates of the cell' is one of the most picturesque, but also one of the most appropriate. Viruses have been known for a long time to utilize a variety of strategies to penetrate cells and, once inside, to take over the host nucleic acid and protein synthesis machinery to build up their own components and produce large amounts of viral progeny. As their genomes carry a minimal amount of information, encoding only a few structural and regulatory proteins, viruses are largely dependent on their hosts for survival; however, despite their apparent simplicity, viruses have evolved different replicative strategies that are regulated in a sophisticated manner. During the last years, the study of the elaborate relationship between viruses and their hosts has led to the understanding of how viral pathogens not only are able to alter the host metabolism via their signaling proteins, but are also able to hijack cellular signaling pathways and transcription factors, and control them to their own advantage. In particular, the nuclear factor-kappaB (NF-kappaB) pathway appears to be an attractive target for common human viral pathogens. This review summarizes what is known about the control of NF-kappaB by viruses, and discusses the possible outcome of NF-kappaB activation during viral infection, which may benefit either the host or the pathogen. PMID- 12773373 TI - Insights into ssDNA recognition by the OB fold from a structural and thermodynamic study of Sulfolobus SSB protein. AB - Information processing pathways such as DNA replication are conserved in eukaryotes and archaea and are significantly different from those found in bacteria. Single-stranded DNA-binding (SSB) proteins (or replication protein A, RPA, in eukaryotes) play a central role in many of these pathways. However, whilst euryarchaea have a eukaryotic-type RPA homologue, crenarchaeal SSB proteins appear much more similar to the bacterial proteins, with a single OB fold for DNA binding and a flexible C-terminal tail that is implicated in protein protein interactions. We have determined the crystal structure of the SSB protein from the crenarchaeote Sulfolobus solfataricus to 1.26 A. The structure shows a striking and unexpected similarity to the DNA-binding domains of human RPA, providing confirmation of the close relationship between archaea and eukaryotes. The high resolution of the structure, together with thermodynamic and mutational studies of DNA binding, allow us to propose a molecular basis for DNA binding and define the features required for eukaryotic and archaeal OB folds. PMID- 12773375 TI - Structure of Rhodococcus erythropolis limonene-1,2-epoxide hydrolase reveals a novel active site. AB - Epoxide hydrolases are essential for the processing of epoxide-containing compounds in detoxification or metabolism. The classic epoxide hydrolases have an alpha/beta hydrolase fold and act via a two-step reaction mechanism including an enzyme-substrate intermediate. We report here the structure of the limonene-1,2 epoxide hydrolase from Rhodococcus erythropolis, solved using single-wavelength anomalous dispersion from a selenomethionine-substituted protein and refined at 1.2 A resolution. This enzyme represents a completely different structure and a novel one-step mechanism. The fold features a highly curved six-stranded mixed beta-sheet, with four alpha-helices packed onto it to create a deep pocket. Although most residues lining this pocket are hydrophobic, a cluster of polar groups, including an Asp-Arg-Asp triad, interact at its deepest point. Site directed mutagenesis supports the conclusion that this is the active site. Further, a 1.7 A resolution structure shows the inhibitor valpromide bound at this position, with its polar atoms interacting directly with the residues of the triad. We suggest that several bacterial proteins of currently unknown function will share this structure and, in some cases, catalytic properties. PMID- 12773374 TI - Structural basis for SH3 domain-mediated high-affinity binding between Mona/Gads and SLP-76. AB - SH3 domains are protein recognition modules within many adaptors and enzymes. With more than 500 SH3 domains in the human genome, binding selectivity is a key issue in understanding the molecular basis of SH3 domain interactions. The Grb2 like adaptor protein Mona/Gads associates stably with the T-cell receptor signal transducer SLP-76. The crystal structure of a complex between the C-terminal SH3 domain (SH3C) of Mona/Gads and a SLP-76 peptide has now been solved to 1.7 A. The peptide lacks the canonical SH3 domain binding motif P-x-x-P and does not form a frequently observed poly-proline type II helix. Instead, it adopts a clamp-like shape around the circumfence of the SH3C beta-barrel. The central R-x-x-K motif of the peptide forms a 3(10) helix and inserts into a negatively charged double pocket on the SH3C while several other residues complement binding through hydrophobic interactions, creating a short linear SH3C binding epitope of uniquely high affinity. Interestingly, the SH3C displays ion-dependent dimerization in the crystal and in solution, suggesting a novel mechanism for the regulation of SH3 domain functions. PMID- 12773376 TI - Crystal structure of tRNA(m1G37)methyltransferase: insights into tRNA recognition. AB - tRNA(m(1)G37)methyltransferase (TrmD) catalyzes the transfer of a methyl group from S-adenosyl-L- methionine (AdoMet) to G(37) within a subset of bacterial tRNA species, which have a G residue at the 36th position. The modified guanosine is adjacent to and 3' of the anticodon and is essential for the maintenance of the correct reading frame during translation. Here we report four crystal structures of TrmD from Haemophilus influenzae, as binary complexes with either AdoMet or S adenosyl-L-homocysteine (AdoHcy), as a ternary complex with AdoHcy and phosphate, and as an apo form. This first structure of TrmD indicates that it functions as a dimer. It also suggests the binding mode of G(36)G(37) in the active site of TrmD and the catalytic mechanism. The N-terminal domain has a trefoil knot, in which AdoMet or AdoHcy is bound in a novel, bent conformation. The C-terminal domain shows structural similarity to trp repressor. We propose a plausible model for the TrmD(2)-tRNA(2) complex, which provides insights into recognition of the general tRNA structure by TrmD. PMID- 12773377 TI - Structures of immature flavivirus particles. AB - Structures of prM-containing dengue and yellow fever virus particles were determined to 16 and 25 A resolution, respectively, by cryoelectron microscopy and image reconstruction techniques. The closely similar structures show 60 icosahedrally organized trimeric spikes on the particle surface. Each spike consists of three prM:E heterodimers, where E is an envelope glycoprotein and prM is the precursor to the membrane protein M. The pre-peptide components of the prM proteins in each spike cover the fusion peptides at the distal ends of the E glycoproteins in a manner similar to the organization of the glycoproteins in the alphavirus spikes. Each heterodimer is associated with an E and a prM transmembrane density. These transmembrane densities represent either an EE or prMprM antiparallel coiled coil by which each protein spans the membrane twice, leaving the C-terminus of each protein on the exterior of the viral membrane, consistent with the predicted membrane-spanning domains of the unprocessed polyprotein. PMID- 12773378 TI - A reducing system of the superoxide sensor SoxR in Escherichia coli. AB - The soxRS regulon functions in protecting Escherichia coli cells against superoxide and nitric oxide. When SoxR is activated by oxidation of its [2Fe-2S] cluster, it increases the synthesis of SoxS, which then activates its target gene expression. How the oxidized SoxR returns to and is maintained in its reduced state has been under question. To identity genes that constitute the SoxR reducing system, we screened an E.coli mutant library carrying a chromosomal soxSp::lacZ fusion, for constitutive mutants. Mutations mapped to two loci: the rsxABCDGE operon (named for reducer of SoxR) that is highly homologous to the rnfABCDGE operon in Rhodobacter capsulatus involved in transferring electrons to nitrogenase, and the rseC gene in the rpoE-rseABC operon. In-frame deletion of each open reading frame in the rsxABCDGE operon produced a similar constitutive phenotype. The double mutation of rsx and rseC suggested that rsxABCDGE and rseC gene products act together in the same pathway in reducing SoxR. Electron paramagnetic resonance analysis of SoxR and measurement of re-reduction kinetics support the proposal that rsx and rseC gene products constitute a reducing system for SoxR. PMID- 12773379 TI - NADPH oxidase AtrbohD and AtrbohF genes function in ROS-dependent ABA signaling in Arabidopsis. AB - Reactive oxygen species (ROS) have been proposed to function as second messengers in abscisic acid (ABA) signaling in guard cells. However, the question whether ROS production is indeed required for ABA signal transduction in vivo has not yet been addressed, and the molecular mechanisms mediating ROS production during ABA signaling remain unknown. Here, we report identification of two partially redundant Arabidopsis guard cell-expressed NADPH oxidase catalytic subunit genes, AtrbohD and AtrbohF, in which gene disruption impairs ABA signaling. atrbohD/F double mutations impair ABA-induced stomatal closing, ABA promotion of ROS production, ABA-induced cytosolic Ca(2+) increases and ABA- activation of plasma membrane Ca(2+)-permeable channels in guard cells. Exogenous H(2)O(2) rescues both Ca(2+) channel activation and stomatal closing in atrbohD/F. ABA inhibition of seed germination and root elongation are impaired in atrbohD/F, suggesting more general roles for ROS and NADPH oxidases in ABA signaling. These data provide direct molecular genetic and cell biological evidence that ROS are rate limiting second messengers in ABA signaling, and that the AtrbohD and AtrbohF NADPH oxidases function in guard cell ABA signal transduction. PMID- 12773380 TI - Role of the CD47-SHPS-1 system in regulation of cell migration. AB - SHPS-1 is a transmembrane protein whose extracellular region interacts with CD47 and whose cytoplasmic region undergoes tyrosine phosphorylation and there by binds the protein tyrosine phosphatase SHP-2. Formation of this complex is implicated in regulation of cell migration by an unknown mechanism. A CD47-Fc fusion protein or antibodies to SHPS-1 inhibited migration of human melanoma cells or of CHO cells overexpressing SHPS-1. Overexpression of wild-type SHPS-1 promoted CHO cell migration, whereas expression of the SHPS-1-4F mutant, which lacks the phosphorylation sites required for SHP-2 binding, had no effect. Antibodies to SHPS-1 failed to inhibit migration of CHO cells expressing SHPS-1 4F. SHPS-1 ligands induced the dephosphorylation of SHPS-1 and dissociation of SHP-2. Antibodies to SHPS-1 also enhanced Rho activity and induced both formation of stress fibers and adoption of a less polarized morphology in melanoma cells. Our results suggest that engagement of SHPS-1 by CD47 prevents the positive regulation of cell migration by this protein. The CD47- SHPS-1 system and SHP-2 might thus contribute to the inhibition of cell migration by cell-cell contact. PMID- 12773381 TI - Rabaptin-5alpha/rabaptin-4 serves as a linker between rab4 and gamma(1)-adaptin in membrane recycling from endosomes. AB - Rab4 regulates recycling from early endosomes. We investigated the role of the rab4 effector rabaptin-5alpha and its putative partner gamma(1)-adaptin in membrane recycling. We found that rabaptin-5alpha forms a ternary complex with the gamma(1)-sigma(1) subcomplex of AP-1, via a direct interaction with the gamma(1)-subunit. The binding site for gamma(1)-adaptin is in the hinge region of rabaptin-5alpha, which is distinct from rab4- and rab5-binding domains. Endogenous or ectopically expressed gamma(1)- adaptin localized to both the trans Golgi network and endosomes. Co-expressed rabaptin-5alpha and gamma(1)-adaptin, however, co-localized in a rab4-dependent manner on recycling endosomes. Transfection of rabaptin-5alpha caused enlarged endosomes and delayed recycling of transferrin. RNAi of rab4 had an opposing effect on transferrin recycling. Collectively, our data show that rab4-GTP acts as a scaffold for a rabaptin 5alpha- gamma(1)-adaptin complex on recycling endosomes and that interactions between rab4, rabaptin-5alpha and gamma(1)-adaptin regulate membrane recycling. PMID- 12773382 TI - A cholesterol-regulated PP2A/HePTP complex with dual specificity ERK1/2 phosphatase activity. AB - The acute depletion of membrane cholesterol causes the concentration of pERK1/2 in caveola/raft lipid domains and the cytosol of human fibroblasts to dramatically increase. This increase could be caused by either the activation of MEK-1 or the inhibition of a pERK phosphatase. Here we describe the isolation of a high molecular weight ( approximately 440 kDa), cholesterol-regulated pERK phosphatase that dephosphorylates both the phosphotyrosine and the phosphothreonine residues in the activation loop of the enzyme. The dual activity in the complex appears to be due to the combined activities of the serine/threonine phosphatase PP2A and the tyrosine phosphatase HePTP. Acute depletion of cholesterol causes the disassembly of the complex and a concomitant loss of the dual specificity pERK phosphatase activity. The existence of a cholesterol-regulated HePTP/PP2A activity provides a molecular explanation for why ERK activity is sensitive to membrane cholesterol levels, and raises the possibility that ERK plays a role in regulating the traffic of cholesterol to caveolae/rafts and other membranes. PMID- 12773383 TI - Haploinsufficiency-based large-scale forward genetic analysis of filamentous growth in the diploid human fungal pathogen C.albicans. AB - Candida albicans is the most prevalent human fungal pathogen. Here, we take advantage of haploinsufficiency and transposon mutagenesis to perform large-scale loss-of-function genetic screen in this organism. We identified mutations in 146 genes that affect the switch between its single-cell (yeast) form and filamentous forms of growth; this switch appears central to the virulence of C.albicans. The encoded proteins include those involved in nutrient sensing, signal transduction, transcriptional control, cytoskeletal organization and cell wall construction. Approximately one-third of the genes identified in the screen lack homologs in Saccharomyces cerevisiae and other model organisms and thus constitute candidate antifungal drug targets. These results illustrate the value of performing forward genetic studies in bona fide pathogens. PMID- 12773384 TI - TCGAP, a multidomain Rho GTPase-activating protein involved in insulin-stimulated glucose transport. AB - Insulin stimulates glucose uptake in fat and muscle cells via the translocation of the GLUT4 glucose transporter from intracellular storage vesicles to the cell surface. The signaling pathways linking the insulin receptor to GLUT4 translocation in adipocytes involve activation of the Rho family GTPases TC10alpha and beta. We report here the identification of TCGAP, a potential effector for Rho family GTPases. TCGAP consists of N-terminal PX and SH3 domains, a central Rho GAP domain and multiple proline-rich regions in the C-terminus. TCGAP specifically interacts with cdc42 and TC10beta through its GAP domain. Although it has GAP activity in vitro, TCGAP is not active as a GAP in intact cells. TCGAP translocates to the plasma membrane in response to insulin in adipocytes. The N-terminal PX domain interacts specifically with phos phatidylinositol-(4,5)-bisphosphate. Overexpression of the full-length and C terminal fragments of TCGAP inhibits insulin-stimulated glucose uptake and GLUT4 translocation. Thus, TCGAP may act as a downstream effector of TC10 in the regulation of insulin-stimulated glucose transport. PMID- 12773385 TI - Glycoprotein hormone receptors: determinants in leucine-rich repeats responsible for ligand specificity. AB - Glycoprotein hormone receptors [thyrotropin (TSHr), luteinizing hormone/chorionic gonadotropin (LH/CGr), follicle stimulating hormone (FSHr)] are rhodopsin-like G protein-coupled receptors with a large extracellular N-terminal portion responsible for hormone recognition and binding. In structural models, this ectodomain is composed of two cysteine clusters flanking nine leucine-rich repeats (LRRs). The LRRs form a succession of beta-strands and alpha-helices organized into a horseshoe-shaped structure. It has been proposed that glycoprotein hormones interact with residues of the beta-strands making the concave surface of the horseshoe. Gain-of-function homology scanning of the beta strands of glycoprotein hormone receptors allowed identification of the critical residues responsible for the specificity towards human chorionic gonadotropin (hCG). Substitution of eight or two residues of the LH/CGr into the TSHr or FSHr, respectively, resulted in constructs displaying almost the same affinity and sensitivity for hCG as wild-type LH/CGr. Molecular dynamics simulations and additional site-directed mutagenesis provided a structural rationale for the evolution of binding specificity in this duplicated gene family. PMID- 12773386 TI - Defective valvulogenesis in HB-EGF and TACE-null mice is associated with aberrant BMP signaling. AB - Heparin-binding epidermal growth factor (HB-EGF) and betacellulin (BTC) are activating ligands for EGF receptor (EGFR/ErbB1) and ErbB4. To identify their physiological functions, we disrupted mouse HB-EGF and BTC alleles by homologous recombination. Most HB-EGF(-/-) mice died before weaning, and survivors had enlarged, dysfunctional hearts and reduced lifespans. Although BTC(-/-) mice were viable and fertile and displayed no overt defects, the lifespan of double null HB EGF(-/-)/BTC(-/-) mice was further reduced, apparently due to accelerated heart failure. HB-EGF(-/-) newborns had enlarged and malformed semilunar and atrioventricular heart valves, and hypoplastic, poorly differentiated lungs. Defective cardiac valvulogenesis was the result of abnormal mesenchymal cell proliferation during remodeling, and was associated with dramatic increases in activated Smad1/5/8. Consistent with the phenotype, HB-EGF transcripts were localized to endocardial cells lining the margins of wild-type valves. Similarly defective valvulogenesis was observed in newborn mice lacking EGFR and tumor necrosis factor-alpha converting enzyme (TACE). These results suggest that cardiac valvulogenesis is dependent on EGFR activation by TACE-derived soluble HB EGF, and that EGFR signaling is required to regulate bone morphogenetic protein signaling in this context. PMID- 12773387 TI - Caspase 3 activation is essential for efficient influenza virus propagation. AB - Apoptosis is a hallmark event observed upon infection with many viral pathogens, including influenza A virus. The apoptotic process is executed by a proteolytic system consisting of a family of cysteinyl proteases, termed caspases. Since the consequences of apoptosis induction and caspase activation for the outcome of an influenza virus infection are not clear, we have addressed this issue by interfering with expression or function of a major virus-induced apoptosis effector, caspase 3. Surprisingly, influenza virus propagation was strongly impaired in the presence of an inhibitor that blocks caspase 3 and in cells where caspase 3 was partially knocked down by small interfering RNAs. Consistent with these findings, poor replication efficiencies of influenza A viruses in cells deficient for caspase 3 could be boosted 30-fold by ectopic expression of the protein. Mechanistically, the block in virus propagation appeared to be due to retention of the viral RNP complexes in the nucleus, preventing formation of progeny virus particles. Our findings indicate that caspase 3 activation during the onset of apoptosis is a crucial event for efficient influenza virus propagation. PMID- 12773388 TI - HBXIP functions as a cofactor of survivin in apoptosis suppression. AB - Survivin is an anti-apoptotic protein that is overexpressed in most human cancers. We show that survivin forms complexes with a cellular protein, hepatitis B X-interacting protein (HBXIP), which was originally recognized for its association with the X protein of hepatitis B virus (HBX). Survivin-HBXIP complexes, but neither survivin nor HBXIP individually, bind pro-caspase-9, preventing its recruitment to Apaf1, and thereby selectively suppressing apoptosis initiated via the mitochondria/cytochrome c pathway. Viral HBX protein also interacts with the survivin- HBXIP complex and suppresses caspase activation in a survivin-dependent manner. Thus, HBXIP functions as a cofactor for survivin, and serves as a link between the cellular apoptosis machinery and a viral pathogen involved in hepatocellular carcinogenesis. PMID- 12773389 TI - The magnitude of hedgehog signaling activity defines skin tumor phenotype. AB - Gain-of-function mutations in SMO have been implicated in constitutive activation of the hedgehog signaling pathway in human basal cell carcinomas (BCCs). We used a truncated keratin 5 (DeltaK5) promoter to assess the potential role of the human M2SMO mutant in BCC development in adult transgenic mice. DeltaK5-M2SMO mouse epidermis is hyperproliferative, ex presses BCC protein markers and gives rise to numerous epithelial downgrowths invading the underlying dermis. Lesions strikingly similar to human basaloid follicular hamartomas develop, but BCCs do not arise even in elderly mice. Hedgehog target gene transcripts were only modestly upregulated in mouse and human follicular hamartomas, in contrast to the high levels detected in BCCs. Cyclins D1 and D2 were selectively upregulated in mouse BCCs. Our data suggest that the levels of hedgehog pathway activation and G(1) cyclins are major determinants of tumor phenotype in skin, and strongly implicate deregulated hedgehog signaling in the genesis of human basaloid follicular hamartomas. Expression of an activated SMO mutant in keratinocytes appears to be insufficient for the development and/or maintenance of full-blown BCCs. PMID- 12773390 TI - The role of Ppe1/PP6 phosphatase for equal chromosome segregation in fission yeast kinetochore. AB - Mis12 is a kinetochore protein essential for equal chromosome segregation and is evolutionarily conserved from yeast to human. In this study, we report the isolation and characterization of suppressors of the mis12 mutant in fission yeast. Our results indicate that Mis12 is negatively regulated by a highly conserved protein phosphatase Ppe1 (scSit4/dmPPV/hPP6) or its bound partner Ekc1 (scSAP), and it is positively regulated by a counteracting kinase Gsk3. Mass spectrometry analysis shows that at least two sites in Mis12 are phosphorylated. This mechanism of suppression occurs at the level of localization recovery of Mis12 to the kinetochore chromatin. Consistently, Mis12 and a subpopulation of Ppe1/Ekc1 were found to behave like non-histone-type chromatin-associating proteins in the chromatin fractionation assay. Mutant analysis of Ppe1 and Ekc1 revealed that they are important for faithful chromosome segregation, as the mutants exhibited unequal chromosome segregation similar to mis12 in the presence of a low concentration of tubulin poison. Ppe1/PP6 directly or indirectly modulates kinetochore chromatin protein Mis12 to ensure progression into normal anaphase. PMID- 12773391 TI - Condensin but not cohesin SMC heterodimer induces DNA reannealing through protein protein assembly. AB - Condensin and cohesin are chromosomal protein complexes required for chromosome condensation and sister chromatid cohesion, respectively. They commonly contain the SMC (structural maintenance of chromosomes) subunits consisting of a long coiled-coil with the terminal globular domains and the central hinge. Condensin and cohesin holo-complexes contain three and two non-SMC subunits, respectively. In this study, DNA interaction with cohesin and condensin complexes purified from fission yeast was investigated. The DNA reannealing activity is strong for condensin SMC heterodimer but weak for holo-condensin, whereas no annealing activity is found for cohesin heterodimer SMC and Rad21-bound heterotrimer complexes. One set of globular domains of the same condensin SMC is essential for the DNA reannealing activity. In addition, the coiled-coil and hinge region of another SMC are needed. Atomic force microscopy discloses the molecular events of DNA reannealing. SMC assembly that occurs on reannealing DNA seems to be a necessary intermediary step. SMC is eliminated from the completed double-stranded DNA. The ability of heterodimeric SMC to reanneal DNA may be regulated in vivo possibly through the non-SMC heterotrimeric complex. PMID- 12773392 TI - Alp13, an MRG family protein, is a component of fission yeast Clr6 histone deacetylase required for genomic integrity. AB - The post-translational modifications of histones are key to the modulation of chromatin structure. Distinct patterns of modifications established by histone modifying enzymes control diverse chromosomal processes. Here, we report the purification and molecular characterization of the fission yeast Clr6 histone deacetyl ase involved in higher order chromatin assembly. We show that a chromodomain protein Alp13, which belongs to the conserved MRG protein family linked to cellular senescence in humans, is associated with Clr6. In addition, Clr6 interacts with homologs of the mammalian transcriptional co-repressors Sin3, Pst1 and Pst2, and a WD40 repeat-containing protein, Prw1. Alp13, Pst2 and Prw1 form a stable complex with Clr6 in the nucleus. Deletion of any of these factors causes progressive loss of viability and sensitivity to DNA-damaging agents, and impairs condensation/resolution of chromosomes during mitosis. This is accompanied by hyperacetylation of histones and a reduction in histone H3 Ser10 phosphorylation, which correlates with chromosome condensation during mitosis. These results link the MRG family protein Alp13 to histone deacetylation, and suggest that Clr6 and its associated factors are essential for fundamental chromosomal events. PMID- 12773393 TI - MSK2 and MSK1 mediate the mitogen- and stress-induced phosphorylation of histone H3 and HMG-14. AB - Cells respond to mitogenic or stress stimuli by the rapid induction of immediate early (IE) genes, which occurs concomitantly with the phosphorylation of histone H3 and the high-mobility-group protein HMG-14. In mammalian cells this response is mediated via ERK and p38 MAP kinase pathways, but the identity of the downstream kinase that phosphorylates histone H3 has been contentious. One study, based on Coffin- Lowry cells defective in RSK2, reported that RSK2 was the histone H3 kinase, while a second study, based on the efficiency of RSKs and MSKs as in vitro histone H3 kinases, and their relative susceptibility to kinase inhibitors, suggested that MSKs were responsible. We show here that the histone H3 phosphorylation response is normal in Coffin-Lowry cells. Further more, we show that histone H3 and HMG-14 phosphorylation is severely reduced or abolished in mice lacking MSK1 and MSK2. We also show that, despite this, histone H3 acetylation is unimpaired in these cells and that IE genes can be induced, although at a reduced efficiency. We conclude that MSKs are the major kinases for histone H3 and HMG-14 in response to mitogenic and stress stimuli in fibroblasts. PMID- 12773394 TI - Epigenetic consequences of AML1-ETO action at the human c-FMS locus. AB - Although many leukaemia-associated nuclear oncogenes are well characterized, little is known about the molecular details of how they alter gene expression. Here we examined transcription factor complexes and chromatin structure of the human c-FMS gene in normal and leukaemic cells. We demonstrate by in vivo footprinting and chromatin immunoprecipitation assays that this gene is bound by the transcription factor AML1 (RUNX1). In t(8;21) leukaemic cells expressing the aberrant fusion protein AML1-ETO, we demonstrate that this protein is part of a transcription factor complex binding to extended sequences of the c-FMS intronic regulatory region rather than the promoter. The AML1-ETO complex does not disrupt binding of other transcription factors, indicating that c-FMS is not irreversibly epigenetically silenced. However, AML1-ETO binding correlates with changes in the histone modification pattern and increased association of histone deacetylases. Our experiments provide for the first time a direct insight into the chromatin structure of an AML1-ETO-bound target gene. PMID- 12773395 TI - p53 represses RNA polymerase III transcription by targeting TBP and inhibiting promoter occupancy by TFIIIB. AB - The tumor suppressor p53 is a transcription factor that controls cellular growth and proliferation. p53 targets include RNA polymerase (pol) III-dependent genes encoding untranslated RNAs such as tRNA and 5S rRNA. These genes are repressed through interaction of p53 with TFIIIB, a TATA-binding protein (TBP)-containing factor. Although many studies have shown that p53 binds to TBP, the significance of this interaction has remained elusive. Here we demonstrate that the TBP-p53 interaction is of functional importance for regulating RNA pol III-transcribed genes. Unlike RNA pol II-dependent promoter repression, overexpressing TBP can reverse inhibition of tRNA gene transcription by p53. p53 does not disrupt the direct interaction between the TFIIIB subunits TBP and Brf1, but prevents the association of Brf1 complexes with TFIIIC2 and RNA pol III. Using chromatin immunoprecipitation assays, we found that TFIIIB occupancy on tRNA genes markedly decreases following p53 induction, whereas binding of TFIIIC2 to these genes is unaffected. Together our results support the idea that p53 represses RNA pol III transcription through direct interactions with TBP, preventing promoter occupancy by TFIIIB. PMID- 12773396 TI - Recognition of GU-rich polyadenylation regulatory elements by human CstF-64 protein. AB - Vertebrate polyadenylation sites are identified by the AAUAAA signal and by GU rich sequences downstream of the cleavage site. These are recognized by a heterotrimeric protein complex (CstF) through its 64 kDa subunit (CstF-64); the strength of this interaction affects the efficiency of poly(A) site utilization. We present the structure of the RNA-binding domain of CstF-64 containing an RNA recognition motif (RRM) augmented by N- and C-terminal helices. The C-terminal helix unfolds upon RNA binding and extends into the hinge domain where interactions with factors responsible for assembly of the polyadenylation complex occur. We propose that this conformational change initiates assembly. Consecutive Us are required for a strong CstF-GU interaction and we show how UU dinucleotides are recognized. Contacts outside the UU pocket fine tune the protein-RNA interaction and provide different affinities for distinct GU-rich elements. The protein-RNA interface remains mobile, most likely a requirement to bind many GU rich sequences and yet discriminate against other RNAs. The structural distinction between sequences that form stable and unstable complexes provides an operational distinction between weakly and strongly processed poly(A) sites. PMID- 12773397 TI - Pti1p and Ref2p found in association with the mRNA 3' end formation complex direct snoRNA maturation. AB - Eukaryotic RNA polymerase II transcribes precursors of mRNAs and of non-protein coding RNAs such as snRNAs and snoRNAs. These RNAs have to be processed at their 3' ends to be functional. mRNAs are matured by cleavage and polyadenylation that require a well-characterized protein complex. Small RNAs are also subject to 3' end cleavage but are not polyadenylated. Here we show that two newly identified proteins, Pti1p and Ref2p, although they were found associated with the pre-mRNA 3' end processing complex, are essential for yeast snoRNA 3' end maturation. We also provide evidence that Pti1p probably acts by uncoupling cleavage and polyadenylation, and functions in coordination with the Nrd1p-dependent pathway for 3' end formation of non-polyadenylated transcripts. PMID- 12773399 TI - The genome-linked protein VPg of the Norwalk virus binds eIF3, suggesting its role in translation initiation complex recruitment. AB - The positive-strand RNA genomes of caliciviruses are not capped, but are instead covalently linked at their 5' ends to a viral protein called VPg. The lack of a cap structure typical of eukaryotic mRNA and absence of an internal ribosomal entry site suggest that VPg may function in translation initiation on calicivirus RNA. This hypothesis was tested by analyzing binding of Norwalk virus VPg to translation initiation factors. The eIF3d subunit of eIF3 was identified as a binding partner of VPg by yeast two-hybrid analysis. VPg bound to purified mammalian eIF3 and to eIF3 in mammalian cell lysates. To test the effects of the VPg- eIF3 interaction on translation, VPg was added to cell-free translation reactions programmed with either capped reporter RNA, an RNA containing an EMCV internal ribosomal entry site (IRES) or an RNA with a cricket paralysis virus IRES. VPg inhibited translation of all reporter RNAs in a dose-dependent manner. Together, the data suggest that VPg may play a role in initiating translation on calicivirus RNA through unique protein-protein interactions with the translation machinery. PMID- 12773398 TI - Coordinated nuclear export of 60S ribosomal subunits and NMD3 in vertebrates. AB - 60S and 40S ribosomal subunits are assembled in the nucleolus and exported from the nucleus to the cytoplasm independently of each other. We show that in vertebrate cells, transport of both subunits requires the export receptor CRM1 and Ran.GTP. Export of 60S subunits is coupled with that of the nucleo- cytoplasmic shuttling protein NMD3. Human NMD3 (hNMD3) contains a CRM-1-dependent leucine-rich nuclear export signal (NES) and a complex, dispersed nuclear localization signal (NLS), the basic region of which is also required for nucleolar accumulation. When present in Xenopus oocytes, both wild-type and export-defective mutant hNMD3 proteins bind to newly made nuclear 60S pre-export particles at a late step of subunit maturation. The export-defective hNMD3, but not the wild-type protein, inhibits export of 60S subunits from oocyte nuclei. These results indicate that the NES mutant protein competes with endogenous wild type frog NMD3 for binding to nascent 60S subunits, thereby preventing their export. We propose that NMD3 acts as an adaptor for CRM1-Ran.GTP-mediated 60S subunit export, by a mechanism that is conserved from vertebrates to yeast. PMID- 12773401 TI - Expression pattern of metalloproteinases and tissue inhibitors of matrix metalloproteinases in cycling human endometrium. AB - The cyclic growth, differentiation, and cell death of endometrium represents the most dynamic example of steroid-driven tissue turnover in human adults. Key effectors in these processes-matrix metalloproteinases (MMPs) and their specific inhibitors (TIMPs)-are regulated by ovarian steroids and, locally, by cytokines. We used reverse transcription-polymerase chain reaction to evaluate the expression of both transcriptionally regulated molecules such as estrogen receptor-alpha, progesterone receptor, and prolactin and a large array of MMPs and TIMPs (MMP-1, -2, -3, -7, -8, -9, -11, -12, -19, -26, MT1-MMP, MT2-MMP, MT3 MMP, TIMP-1, -2, -3). Altogether, three distinct patterns of MMP and two patterns of TIMP expression were detected in cycling endometrium: 1). MMPs restricted to the menstrual period (MMPs-1, -3, -8, -9, -12); 2). MMPs and TIMPs expressed throughout the cycle (MMP-2, MT1-MMP, MT2-MMP, MMP-19, TIMP-1, and TIMP-2); 3). MMPs predominantly expressed during the proliferative phase (MMP-7, MMP-11, MMP 26, and MT3-MMP); and 4). TIMP-3, which, contrary to the other TIMPs, shows significant modulations, with maximum expression during the late secretory and menstrual phases. These specific patterns of MMP expression associated with each phase of the cycle may point to specific roles in the processes of menstruation, housekeeping activities, angiogenesis, tissue growth, and extracellular matrix remodeling. PMID- 12773400 TI - A subset of ATM- and ATR-dependent phosphorylation events requires the BRCA1 protein. AB - BRCA1 is a central component of the DNA damage response mechanism and defects in BRCA1 confer sensitivity to a broad range of DNA damaging agents. BRCA1 is required for homologous recombination and DNA damage-induced S and G(2)/M phase arrest. We show here that BRCA1 is required for ATM- and ATR-dependent phosphorylation of p53, c-Jun, Nbs1 and Chk2 following exposure to ionizing or ultraviolet radiation, respectively, and is also required for ATM phosphorylation of CtIP. In contrast, DNA damage-induced phosphorylation of the histone variant H2AX is independent of BRCA1. We also show that the presence of BRCA1 is dispensable for DNA damage-induced phosphorylation of Rad9, Hus1 and Rad17, and for the relocalization of Rad9 and Hus1. We propose that BRCA1 facilitates the ability of ATM and ATR to phosphorylate downstream substrates that directly influence cell cycle checkpoint arrest and apoptosis, but that BRCA1 is dispensable for the phosphorylation of DNA-associated ATM and ATR substrates. PMID- 12773402 TI - Human oocytes reversibly arrested in prophase I by phosphodiesterase type 3 inhibitor in vitro. AB - This study addresses the role of cAMP hydrolytic isoenzyme phosphodiesterase type 3 (PDE 3) modulation on human oocyte maturation in vitro. Presence of phosphodiesterase type 3 A (PDE 3A) mRNA was confirmed in human germinal vesicle stage (GV) oocytes. Making use of a selective PDE 3 inhibitor, Org 9935 (10 microM), oocytes retrieved from immature follicles were arrested in prophase I with a high efficiency for up to 72 h. Cumulus oocyte complexes (COCs) were retrieved in the follicular phase of the cycle before or after exposure to endogenous LH or hCG administration in vivo and randomly distributed into maturation medium with or without the PDE 3 inhibitor. Previous exposure of small follicles to LH activity in vivo had no influence on the arresting capacity of the PDE 3 inhibitor. Reversal from pharmacological arrest leads to a progression through meiosis in a normal time frame with formation of a well-aligned metaphase plate. Ultrastructure analysis of COC derived from follicles between 8 and 12 mm showed that the induced extension of prophase I arrest in vitro resulted in cytoplasm changes but not in apparent nuclear changes during culture. PMID- 12773403 TI - Zygote arrest 1 (Zar1) is an evolutionarily conserved gene expressed in vertebrate ovaries. AB - Zygote arrest 1 (ZAR1) is an ovary-specific maternal factor that plays essential roles during the oocyte-to-embryo transition. In mice, the Zar1 mRNA is detected as a 1.4-kilobase (kb) transcript that is synthesized exclusively in growing oocytes. To further understand the functions of ZAR1, we have cloned the orthologous Zar1 cDNA and/or genes for mouse, rat, human, frog, zebrafish, and pufferfish. The entire mouse Zar1 gene and a related pseudogene span approximately 4.0 kb, contain four exons, and map to adjacent loci on mouse chromosome 5. The human ZAR1 orthologous gene similarly consists of four exons and resides on human chromosome 4p12, which is syntenic with the mouse Zar1 chromosomal locus. Rat (Rattus norvegicus) and pufferfish (Fugu rubripes) Zar1 genes were recognized by database mining and deduced protein alignment analysis. The rat Zar1 gene also maps to a region that is syntenic with the mouse Zar1 gene locus on rat chromosome 14. Frog (Xenopus laevis) and zebrafish (Danio rerio) Zar1 orthologs were cloned by reverse transcription-polymerase chain reaction and rapid amplification of cDNA ends analysis of ovarian mRNA. Unlike mouse and human, the frog Zar1 is detected in multiple tissues, including lung, muscle, and ovary. The Zar1 mRNA appears in the cytoplasm of oocytes and persists until the tailbud stage during frog embryogenesis. Mouse, rat, human, frog, zebrafish, and pufferfish Zar1 genes encode proteins of 361, 361, 424, 295, 329, and 320 amino acids, respectively, and share 50.8%-88.1% amino acid identity. Regions of the N termini of these ZAR1 orthologs show high sequence identity among these various proteins. However, the C-terminal 103 amino acids of these proteins, encoded by exons 2-4, contain an atypical eight-cysteine Plant Homeo Domain motif and are highly conserved, sharing 80.6%-98.1% identity among these species. These findings suggest that the carboxyl-termini of these ZAR1 proteins contain an important functional domain that is conserved through vertebrate evolution and that may be necessary for normal female reproduction in the transition from oocyte to embryonic life. PMID- 12773404 TI - ESP13.2, a member of the beta-defensin family, is a macaque sperm surface-coating protein involved in the capacitation process. AB - Female macaques produced isoantibodies to a limited number of sperm surface proteins following immunization with sperm components released by phosphatidylinositol-specific phospholipase C (PI-PLC). Washed, acrosome-intact, fixed sperm injected into rabbits elicited a major immune response to one of the same PI-PLC-released proteins, which was shown to be a sperm surface-coating protein. After purification and digestion of the glycoprotein, four peptides were analyzed for amino acid sequence, and all had 100% homology with an epididymal secretory protein, ESP13.2, reported previously to be a small, cationic-rich peptide and a member of the beta-defensin family. Antibodies to purified ESP13.2 recognized a number of protein bands on Western blots of nonreduced PI-PLC released sperm components and nonreduced whole-sperm extracts. After chemical disulfide reduction, only a single, broad band from 31 to 35 kDa was recognized by anti-ESP13.2 antibodies. Indirect immunofluorescence showed ESP13.2 over the entire surface of ejaculated macaque sperm. Fluorescence was only slightly reduced after sperm were washed through 80% Percoll. A 24-h incubation in capacitating medium significantly reduced the amount of ESP13.2 over the head and midpiece, whereas exposure of the incubated sperm to dbcAMP and caffeine (capacitation activators) resulted in almost complete loss of ESP13.2 from the sperm surface. After activation, ESP13.2 was the primary component released into the medium as judged electrophoretically. Lignosulfonic acid, a potent inhibitor of macaque fertilization in vitro, completely blocked release of ESP13.2 from the sperm surface, even following treatment with activators. These findings suggest that the beta-defensin, ESP13.2, has a function in the capacitation of macaque spermatozoa and may modulate sperm surface-receptor presentation at the time of fertilization. PMID- 12773405 TI - Numerical chromosomal abnormalities in rat epididymal spermatozoa following chronic cyclophosphamide exposure. AB - Chronic, low-dose treatment of male rats with cyclophosphamide, a chemotherapeutic agent, is known to affect progeny outcome adversely in a dose dependent and time-specific manner, resulting in increased pre- and postimplantation loss as well as malformations. Concern exists regarding the genetic quality of mature gametes exposed to cyclophosphamide during mitosis and meiosis. The goal of the present study was to determine the effect of chronic cyclophosphamide treatment during spermatogenesis on the frequency of numerical chromosomal anomalies in epididymal spermatozoa. Male rats were treated with either saline or cyclophosphamide (6 mg kg-1 day-1) for 6 or 9 wk, and cauda epididymal spermatozoa were collected. The rat sperm Y-4 fluorescence in situ hybridization assay was used to assess the induction of spermatozoal disomy, nullisomy, and diploidy involving chromosomes Y and 4. The overall frequency of numerically abnormal spermatozoa was elevated approximately 2-fold (P < 0.001) after 9 wk of cyclophosphamide treatment. Exposure for 9 wk, but not for 6 wk, significantly increased the frequency of spermatozoa with chromosome 4 disomy (P < 0.02) and nullisomy (P < 0.05), but disomy Y and diploidy were not significantly increased with treatment compared to corresponding controls. Independent of treatment, only 27% of aneuploid spermatozoa presented with morphological abnormalities, but all diploid spermatozoa were approximately twice the size of normal cells. Thus, cyclophosphamide disrupts meiotic events before pachynema during spermatogenesis, emphasizing the potential for adverse progeny outcomes following genotoxic damage. PMID- 12773406 TI - Seasonal differences in the effect of isolation and restraint stress on the luteinizing hormone response to gonadotropin-releasing hormone in hypothalamopituitary disconnected, gonadectomized rams and ewes. AB - Stress responses are thought to act within the hypothalamopituitary unit to impair the reproductive system, and the sites of action may differ between sexes. The effect of isolation and restraint stress on pituitary responsiveness to GnRH in sheep was investigated, with emphasis on possible sex differences. Experiments were conducted during the breeding season and the nonbreeding season. In both experiments, 125 ng of GnRH was injected i.v. every 2 h into hypothalamopituitary disconnected, gonadectomized rams and ewes on 3 experimental days, with each day divided into two periods. During the second period on Day 2, isolation and restraint stress was imposed for 5.5 h. Plasma concentrations of LH and cortisol were measured in samples of blood collected from the jugular vein. In the second experiment (nonbreeding season), plasma concentrations of epinephrine, norepinephrine, 3,4-dihydroxyphenylalanine, and 3,4-dihydroxyphenylglycol were also measured. In both experiments, there was no effect of isolation and restraint stress on plasma concentrations of cortisol in either sex. During the breeding season, there was no effect of isolation and restraint stress on plasma concentrations of LH in either sex. During the nonbreeding season, the amplitude of the first LH pulse after the commencement of stress was significantly reduced (P < 0.05) in rams and ewes. In the second experiment, during stress there was a significant increase (P < 0.05) in plasma concentrations of epinephrine in rams and ewes and significantly higher (P < 0.05) basal concentrations of norepinephrine in ewes than in rams. These results suggest that in sheep stress reduces responsiveness of the pituitary gland to exogenous GnRH during the nonbreeding season but not during the breeding season, possibly because of mediators of the stress response other than those of the hypothalamus-pituitary adrenal gland axis. PMID- 12773407 TI - Generation and characterization of a transgenic mouse with a functional human TSPY. AB - To generate an animal model that is suitable for the analysis of regulation and expression of human testis-specific protein, Y-encoded TSPY, a transgenic mouse line, TgTSPY9, harboring a complete structural human TSPY gene was generated. Fluorescence in situ hybridization and Southern analyses show that approximately 50 copies of the human TSPY transgene are integrated at a single chromosomal site that maps to the distal long arm of the Y chromosome. The transgene is correctly transcribed and spliced according to the human pattern and is mainly expressed in testicular tissue, with spermatogonia and early primary spermatocytes (leptotene and zygotene) as expressing germ cells. TSPY transgenic mice are phenotypically normal, and spermatogenesis is neither impaired nor enhanced by the human transgene. The present study shows that a human TSPY gene integrated into the mouse genome follows the human expression pattern although murine tspy had lost its function in rodent evolution millions of years ago. PMID- 12773408 TI - Does alligator testis produce estradiol? A comparison of ovarian and testicular aromatase. AB - Testicular secretion of estradiol is necessary for normal spermatogenesis and male reproductive physiology in humans and rodents. The role of estradiol in nonmammalian vertebrates remains unknown, but elevated circulating estradiol has been reported in male lizards, alligators, and various bird species. We have been unable to detect circulating estradiol in male alligators; therefore, we reexamined the question of testicular production of estradiol in alligators using more rigorous assay procedures. A large pool of plasma from a male alligator was extracted and run through an HPLC column. Immunoreactive estradiol-like material eluted coincident with authentic estradiol. By using an ultrasensitive RIA and processing large volumes of male plasma (1000 microl), we were able to measure estradiol. Estradiol in male alligators ranged from 0.23 to 3.14 pg/ml, whereas estradiol in immature female alligators ranged from 14 to 66 pg/ml. Aromatase activity in microsomes from adult alligator ovarian tissue was 36.2 +/- 1.6 pmol mg-1 h-1, whereas activity in testicular microsomes ranged between 0.92 and 2.38 pmol mg-1 h-1. Ovarian aromatase activity was inhibited in a concentration dependent fashion by Fadrozole, but the essentially background activity of testicular aromatase was not inhibited at any concentration of Fadrozole. Likewise, a comparison of alligator testicular and ovarian aromatase mRNA expression gave a similar result: the ovarian expression was 600-fold higher and brain tissue was 10-fold higher than that of the testis. Circulating estradiol in male alligators is probably of extragonadal origin, and the testis produces little if any of this steroid. PMID- 12773409 TI - Equatorial segment protein defines a discrete acrosomal subcompartment persisting throughout acrosomal biogenesis. AB - The equatorial segment of the acrosome underlies the domain of the sperm that fuses with the egg membrane during fertilization. Equatorial segment protein (ESP), a novel 349-amino acid concanavalin-A-binding protein encoded by a two exon gene (SP-ESP) located on chromosome 15 at q22, has been localized to the equatorial segment of ejaculated human sperm. Light microscopic immunofluorescent observations revealed that during acrosome biogenesis ESP first appears in the nascent acrosomal vesicle in early round spermatids and subsequently segregates to the periphery of the expanding acrosomal vesicle, thereby defining a peripheral equatorial segment compartment within flattened acrosomal vesicles and in the acrosomes of early and late cap phase, elongating, and mature spermatids. Electron microscopic examination revealed that ESP segregates to an electron lucent subdomain of the condensing acrosomal matrix in Golgi phase round spermatids and persists in a similar electron-lucent subdomain within cap phase spermatids. Subsequently, ESP was localized to electron-dense regions of the equatorial segment and the expanded equatorial bulb in elongating spermatids and mature sperm. ESP is the earliest known protein to be recognized as a marker for the specification of the equatorial segment, and it allows this region to be traced through all phases of acrosomal biogenesis. Based on these observations, we propose a new model of acrosome biogenesis in which the equatorial segment is defined as a discrete domain within the acrosomal vesicle as early as the Golgi phase of acrosome biogenesis. PMID- 12773410 TI - Mouse preimplantation embryos developed from oocytes injected with round spermatids or spermatozoa have similar but distinct patterns of early messenger RNA expression. AB - Quantitative real-time polymerase chain reaction assay was used to compare the temporal transcriptional activation and mRNA removal for a number of genes in mouse embryos derived by round spermatid injection (ROSI) or intracytoplasmic sperm injection. A number of marker genes with widely different cellular functions were analyzed. Similar patterns of activation were found for the transcription factor Oct 4, the translation initiation factor eukaryotic initiation factor 1A, the L1 ribosomal protein, the chromatin modifying protein histone deacetylase 1, the enzyme hypoxanthine phosphoribosyl transferase, the murine endogenous retrovirus-like element, and the repetitive DNA LINE retrotransposons. Expression of the retrovirus-like mobile element intracisternal A particle, however, was markedly elevated from the two-cell to the blastocyst stages in ROSI embryos. Analyses performed for various paternal mRNAs introduced into the oocyte by the round spermatid, including protamines 1 and 2, transition protein 2, ropporin, and glyceraldehydes 3-phosphate dehydrogenase, revealed all were removed from the preimplantation embryos, albeit with distinct temporal patterns. PMID- 12773412 TI - Possible expansion of "Window of Implantation" in pseudopregnant mice: time of implantation of embryos at different stages of development transferred into the same recipient. AB - Blastocyst implantation and successful establishment of pregnancy require delicate interactions between the embryo and maternal environment. It is believed that the growth of transferred embryos of different ages is synchronized during preimplantation development and that such embryos are implanted in the uterus at the same time. To define the time of synchronization for developing embryos of different ages, embryos at two different stages of development were transferred separately into the oviducts of the same recipient. We then examined the subsequent development of the embryos at various time intervals after transfer. Pronucleus (PN) stage eggs were transferred separately to the right or left oviduct of recipients on Day 0, while eight-cell embryos (8C) were transferred to the other oviduct. For 8C, 5%, 63%, and 74% of transferred embryos were implanted in the uterus at 42, 66, and 90 h posttransfer, respectively. In contrast, none of the transferred PN was implanted until 90 h posttransfer. At 90 h posttransfer, 59% of the PN had successfully implanted. Histological examination revealed that developmental stage of the embryos in both groups synchronized around 162 h posttransfer, even though the implantation was accelerated in 8C compared with PN. Our results indicate that embryos of advanced stage transferred to the oviduct implant in the uterus in advance of younger embryos and that the uterine development is synchronized at the neural plate, presomite stage. Our results strongly suggest that uterine receptivity for implantation is expandable in pseudopregnant mice. PMID- 12773411 TI - A role for tachykinins in female mouse and rat reproductive function. AB - Tachykinins may be involved in reproduction. A reverse transcription-polymerase chain reaction assay was used to analyze the expression of tachykinins and tachykinin receptors in different types of reproductive cells from mice. The preprotachykinin (PPT) genes, PPT-A, PPT-B and PPT-C, that encode substance P/neurokinin A, neurokinin B, and hemokinin-1, respectively, and the genes that encode the tachykinin NK1, NK2, and NK3 receptors were all expressed, at different levels, in the uterus of superovulated, unfertilized mice. The mRNA of neprilysin (NEP), the main enzyme involved in tachykinin metabolism, was also expressed in the uterus. Isolated cumulus granulosa cells expressed PPT-A, PPT-B, PPT-C, and NEP and low levels of the tachykinin NK1 and NK2 receptors. Mouse oocytes expressed PPT-A and -B mRNA transcripts. A low expression of the three tachykinin receptors was observed but PPT-C and NEP were undetectable. Two- and 8 to 16-cell mouse embryos expressed only a low-abundance transcript corresponding to the NK1 receptor. However, the mRNAs of PPT-B, PPT-C and NEP appeared in blastocyst-stage embryos. A low-abundance transcript corresponding to the NK2 receptor was the only target gene detected in mice sperm. Female mice or rats treated neonatally with capsaicin showed a reduced fertility. A reduction in litter size was observed in female rats treated in vivo with the tachykinin NK3 receptor antagonist SR 142801. These data show that tachykinins of both neuronal and nonneuronal origin are differentially expressed in various types of reproductive cells and may play a role in female reproductive function. PMID- 12773413 TI - Generation of live fry from intraperitoneally transplanted primordial germ cells in rainbow trout. AB - Germ cell transplantation has tremendous applications in transgenic animal production, assisted reproductive technology, and germline stem cell research. Here, we report for the first time the production of individuals from intraperitoneally transplanted primordial germ cells (PGCs) in animals. To trace the behavior of exogenous PGCs in recipients, PGCs visualized by a green fluorescent protein gene were used as donors. The PGCs prepared from the genital ridges of hatching embryos were transplanted into recipients at various developmental stages. The PGCs injected into the peritoneal cavities of hatching embryos had the ability to migrate toward, and to colonize, the genital ridges of recipient embryos. Furthermore, donor-derived PGCs proliferated and differentiated into mature eggs and sperm in the allogenic gonads; the resulting gametes produced live fry, showing the donor-derived phenotype, through fertilization. Combined with in vitro culture, genetic modification, and cryopreservation of PGCs, this technique provides new approaches for fish bioengineering. PMID- 12773414 TI - Somatostatin-14 neurons in the ovine hypothalamus: colocalization with estrogen receptor alpha and somatostatin-28(1-12) immunoreactivity, and activation in response to estradiol. AB - Pituitary gland growth hormone (GH) secretion is influenced by two hypothalamic neuropeptides: growth hormone-releasing hormone (GHRH) and somatostatin. Recent data also suggest that estrogen modulates GH release, particularly at the time of the preovulatory luteinizing hormone surge, when a coincident surge of GH is observed in sheep. The GHRH neurons do not possess estrogen receptor alpha (ERalpha), suggesting that estrogen does not act directly on GHRH neurons. Similarly, few somatotropes express ERalpha, suggesting a weak pituitary effect of estradiol on GH. It was hypothesized, therefore, that estradiol may affect somatostatin neurons to modulate GH release from the pituitary. Using immunocytochemical approaches, the present study revealed that although somatostatin neurons were located in several hypothalamic sites, only those in the arcuate nucleus (13% +/- 2%) and ventromedial nucleus (VMN; 29% +/- 1%) expressed ERalpha. In addition, we found that all neurons immunoreactive for somatostatin-14 were also immunoreactive for somatostatin-28(1-12). To determine whether increased GH secretion in response to estradiol is through modulation of GHRH and/or somatostatin neuronal activity, a final study investigated whether c fos expression increased in somatostatin- and GHRH-immunoreactive cells at the time of the estradiol-induced LH surge in intact anestrous ewes. Estradiol significantly (P < 0.05) increased the percentage of GHRH (estradiol, 75% +/- 3%; no estradiol, 19% +/- 2%) neurons expressing c-fos in the hypothalamus. The percentage of somatostatin-immunoreactive neurons coexpressing c-fos in the estradiol-treated animals was significantly (P < 0.05) higher (periventricular, 44% +/- 3%; arcuate, 72% +/- 5%; VMN, 81% +/- 5%) than in the control animals (periventricular, 22% +/- 1%; arcuate, 29% +/- 3%; VMN, 31% +/- 3%). The present study suggests that estradiol modulates the activity of GHRH and somatostatin neurons but that this effect is most likely mediated through an indirect interneuronal pathway. PMID- 12773415 TI - Sodium-inorganic phosphate cotransporter NaPi-IIb in the epididymis and its potential role in male fertility studied in a transgenic mouse model. AB - Analysis by cDNA microarrays showed that in the murine epididymis, NaPi-IIb was the predominantly expressed epithelial isoform of the sodium-inorganic phosphate cotransporter and was markedly overexpressed in the proximal region in the infertile knockout (KO) compared to the fertile heterozygous (HET) c-ros transgenic mouse. The apparent up-regulation in the KO mouse confirmed by Northern and Western blot analyses could be explained by the absence of NaPi-IIb from the initial segment of the HET epididymis, as revealed by immunohistochemistry, and its presence on the epithelial brush border throughout the proximal epididymis of KO mice, where differentiation of the initial segment fails to occur. Both NaPi-IIb mRNA and protein were scarce or absent from the cauda epididymidis of both genotypes. A high content of inorganic phosphate was measured enzymatically in the HET cauda luminal fluid, with a 27% decrease in the KO mice. This decrease, presumably from a greater reabsorption of inorganic phosphate, particularly in the initial part of the KO epididymis, may disturb the normal process of sperm maturation in these infertile males. By contrast, no apparent consequences were observed for the transport of Na+ and Ca2+, the concentrations of which (approximately 26 mM and approximately 30 microM, respectively) were measured by microelectrodes to be identical in the caudal fluid from both genotypes. PMID- 12773416 TI - Ammonium induces aberrant blastocyst differentiation, metabolism, pH regulation, gene expression and subsequently alters fetal development in the mouse. AB - The presence of ammonium in the culture medium has significant detrimental effects on the regulation of embryo physiology and genetics. Ammonium levels build up linearly over time in the culture medium when media containing amino acids are incubated at 37 degrees C. Ammonium in the culture media significantly reduces blastocyst cell number, decreases inner cell mass development, increases apoptosis, perturbs metabolism, impairs the ability of embryos to regulate intracellular pH, and alters the expression of the imprinted gene H19. In contrast, the rate of blastocyst development and blastocyst morphology appear to be normal. The transfer of blastocysts exposed to ammonium results in a significant reduction in the ability to establish a pregnancy. Furthermore, of those embryos that manage to implant, fetal growth is significantly impaired. Embryos exposed to 300 microM ammonium are retarded by 1.5 days developmentally at Day 15 of pregnancy. It is therefore essential that culture conditions for mammalian embryos are designed to minimize the buildup of ammonium to prevent abnormalities in embryo physiology, genetic regulation, pregnancy, and fetal development. PMID- 12773417 TI - Deviation of the blastocyst axis from the first cleavage plane does not affect the quality of mouse postimplantation development. AB - Several researchers have suggested recently that the embryonic-abembryonic (Em Ab) axis of the mouse blastocyst is orthogonal to the first cleavage plane of the two-cell embryo. To determine the universality of this relationship, we used embryos of two different genotypes, F1 (C57BL/6 x DBA/2) and CD-1. The position of the first cleavage plane in the early blastocyst was determined by labeling a blastomere with the fluorescent lineage tracer DiI (1,1'-dioctadecyl-3,3,3',3' tetramethylindocarbocyanine perchlorate) at the two-cell stage. Approximately one quarter of the blastocysts from both genotypes possessed an Em-Ab axis that respected the orthogonal relationship with the first cleavage plane. However, the remainder of the blastocysts deviated from the orthogonal relationship. This result indicates that the orthogonal orientation of the Em-Ab axis to the first cleavage plane is not a universal phenomenon. We also tested whether the angular relationship between the Em-Ab axis and first cleavage plane influences postimplantation embryo development. We sorted the blastocysts that had the Em-Ab axis orthogonal to the first cleavage plane from the ones that did not. These two types of blastocysts were transferred separately into surrogates, and fetal development was examined in late gestation. The results revealed that both types of blastocysts produced normal fetuses at a similar frequency. Thus, the relationship of the blastocyst axis to the first cleavage plane does not significantly influence later development. PMID- 12773418 TI - Production of cloned pigs by whole-cell intracytoplasmic microinjection. AB - Cloning by somatic cell nuclear transfer has been successfully achieved by both fusing of a donor cell with and injecting an isolated donor cell nucleus into an enucleated oocyte. However, each of the above methods involves extended manipulation of either the oocytes (fusion) or the donor cells (nucleus isolation). Additionally, cloning efficiency can be reduced by low fusion rate of the cell fusion method, and specialized micromanipulation equipment and exacting nucleus isolation techniques are required for the nucleus injection method. Here we report a whole-cell injection technique for nuclear transfer in pigs and the production of cloned piglets with comparable, if not higher, efficiency than the other two nuclear transfer procedures. First, we tested the feasibility of this technique with three types of frequently used donor cells (cumulus, mural granulosa, and fibroblasts) and obtained the optimal nuclear reprogramming conditions for these cells. We further improved our protocol by avoiding ultraviolet exposure during enucleation and achieved a 37% blastocyst rate. We then conducted whole-cell injection using skin fibroblasts from the ear of a sow transgenic for two genes, the porcine lactoferrin and the human factor IX, and produced four live-born cloned transgenic piglets from three recipients. The present study demonstrated the applicability of producing normal, cloned piglets by the simple and less labor-intensive whole-cell intracytoplasmic injection. PMID- 12773419 TI - Concentrations of progesterone, follistatin, and follicle-stimulating hormone in peripheral plasma across the estrous cycle and pregnancy in merino ewes that are homozygous or noncarriers of the Booroola gene. AB - The circulating concentrations of progesterone, FSH, and follistatin across the estrous cycle and gestation were compared in Australian merino sheep that were homozygous for the Booroola gene, FecB, or were noncarriers. The Booroola phenotype is due to a point mutation in the bone morphogenetic protein receptor 1B. Progesterone concentrations began to rise earlier and were higher in the Booroola ewes than in the noncarriers on most days of the luteal phase but not during the follicular phase of the cycle. Follistatin concentrations remained unchanged across the estrous cycle in both groups of ewes, with no differences between genotypes. FSH concentrations were higher in Booroola ewes than in noncarrier ewes on most days of the estrous cycle, with a significantly higher and broader peak of FSH around the time of estrus. Progesterone concentrations were significantly higher in early and midgestation in Booroola ewes but were lower toward the end of gestation than those in noncarriers. FSH declined in both groups across gestation, with lower concentrations of FSH in Booroola ewes during midgestation. Follistatin remained unchanged across gestation in Booroola ewes and noncarrier ewes with a twin pregnancy but declined across gestation in noncarrier ewes with a singleton pregnancy. These results suggest that follistatin concentration is not regulated by the FecB gene during the estrous cycle and pregnancy but is influenced by the number of fetuses. However, the FecB gene appears to positively affect both progesterone and FSH during the estrous cycle and across pregnancy, which suggests that bone morphogenetic proteins play an important role in the regulation of both hormones. PMID- 12773420 TI - Developmental and hormonal regulation of the monocarboxylate transporter 2 (MCT2) expression in the mouse germ cells. AB - During spermatogenesis, postmeiotic germ cells utilize lactate produced by Sertoli cells as an energy metabolite. While the hormonal regulation of lactate production in Sertoli cells has been relatively well established, the transport of this energy substrate to the germ cells, particularly via the monocarboxylate transporters (MCTs), as well as the potential endocrine control of such a process remain to be characterized. Here, we report the developmentally and hormonally regulated expression of MCT2 in the testis. At Day 18, MCT2 starts to be expressed in germ cells as detected by Northern blot. The mRNA are translated into protein (40 kDa) in elongating spermatids. Ultrastructural analysis demonstrated that MCT2 protein is localized to the outer face of the cell membrane of spermatid tails. MCT2 mRNA levels are under the control of the endocrine, specifically follicle-stimulating hormone (FSH) and testosterone, and paracrine systems. Indeed, a 35-day-old rat hypophysectomy resulted in an 8-fold increase in testicular MCT2 mRNA levels. Conversely, FSH and LH administration to the hypophysectomized rats reduced MCT2 mRNA levels to the basal levels observed in intact animals. The decrease in MCT2 mRNA levels was confirmed in vitro using isolated seminiferous tubules incubated with FSH or testosterone. FSH or testosterone inhibited in a dose-dependent manner MCT2 mRNA levels with maximal inhibitory doses of 2.2 ng/ml and 55.5 ng/ml for FSH and testosterone, respectively. In addition to the endocrine control, TNFalpha and TGFbeta also exerted an inhibitory effect on MCT2 mRNA levels with a maximal effect at 10 ng/ml and 6.6 ng/ml for TGFbeta and TNFalpha, respectively. Together with previous studies, the present data reinforce the concept that among the key functions of the endocrine/paracrine systems in the testis is the control of the energy metabolism occurring in the context of Sertoli cell-germ cell metabolic cooperation where lactate is produced in somatic cells and transported to germ cells via, at least, MCT2. PMID- 12773421 TI - Acquisition of arylsulfatase A onto the mouse sperm surface during epididymal transit. AB - Arylsulfatase A (AS-A) is localized to the sperm surface and participates in sperm-zona pellucida binding. We investigated how AS-A, usually known as an acrosomal enzyme, trafficked to the sperm surface. Immunocytochemistry of the mouse testis confirmed the existence of AS-A in the acrosomal region of round and elongating spermatids. However, immunofluorescence and flow cytometry indicated the absence of AS-A on the surface of live testicular sperm. In contrast, positive AS-A staining was observed in the heads of live caudal epididymal and vas deferens sperm. The results suggested that acquisition of AS-A on the sperm surface occurred during epididymal transit. Immunocytochemistry of the epididymis revealed AS-A in narrow and apical cells in the initial segment and in clear cells in all epididymal regions. However, these epithelial cells are in the minority and are not involved in secretory activity. In the caudal epididymis and vas deferens, AS-A was also localized to principal cells, the major epithelial cells. Because principal cells have secretory activity, they may secrete AS-A into the epididymal fluid. This hypothesis was supported by our results revealing the presence of AS-A in the epididymal and vas deferens fluid (determined by immunoblotting and ELISA) and an AS-A transcript in the epididymis (by reverse transcription polymerase chain reaction). Alexa-430 AS-A bound to epididymal sperm with high affinity (Kd = 46 nM). This binding was inhibited by treatment of sperm with an antibody against sperm surface sulfogalactosylglycerolipid. This finding suggests that AS-A in the epididymal fluid may deposit onto sperm via its affinity to sulfogalactosylglycerolipid. PMID- 12773422 TI - Chronology of apoptosis in bovine embryos produced in vivo and in vitro. AB - The postimplantation developmental potential of embryos can be affected by various forms of cell death, such as apoptosis, at preimplantation stages. However, correct assessment of apoptosis is needed for adequate inference of the developmental significance of this process. This study is the first to investigate the independent chronological occurrence of apoptotic changes in nuclear morphology and DNA degradation (detected by the TUNEL reaction) and incidences of nuclei displaying these features at various preimplantation stages of bovine embryos produced both in vivo and in vitro. Different elements of apoptosis were observed at various developmental stages and appeared to be differentially affected by in vitro production. Nuclear condensation was observed from the 6-cell stage in vitro and the 8-cell stage in vivo, whereas the TUNEL reaction was first observed at the 6-cell stage in vitro and the 21-cell stage in vivo. Morphological signs of other forms of cell death were also observed in normally developing embryos produced both in vivo and in vitro. The onset of apoptosis seems to be developmentally regulated in a stage-specific manner, but discrete features of the apoptotic process may be differentially regulated and independently modulated by the mode of embryo production. Significant differences in indices of various apoptotic features were not evident between in vivo- and in vitro-produced embryos at the morula stage, but such differences could be observed at the blastocyst stage, where in vitro production was associated with a higher degree of apoptosis in the inner cell mass. PMID- 12773423 TI - Localized expression of angiopoietin 1 and 2 may explain unique characteristics of the rat testicular microvasculature. AB - The testicular vasculature is unique in several ways. The unfenestrated endothelial cells constitute one part of the blood-testis barrier, and testicular microvessels are normally resistant to inflammation mediators. At the same time that angiogenic factors and inflammation mediators are constitutively produced, the proportion of proliferating endothelial cells is considerably higher than in other organs, but new blood vessels are not formed. Hormonal stimulation of the testis with hCG increase endothelial cell proliferation, vascular permeability, and sensitivity to locally injected inflammation mediators. In the present study, we examined whether local expression of angiopoietin (ang) 1, an inhibitor of vascular leakage and sprouting angiogenesis, and its antagonist, ang 2, could be involved in establishing this vascular phenotype. Using reverse transcription polymerase chain reaction and immunohistochemistry, we demonstrate that testicular vascular endothelial growth factor-A (VEGF-A), ang 1, ang 2, and the ang-receptor tie 2 are expressed in the testis and that hormonal stimulation with hCG is accompanied by increased expression of VEGF-A and ang 2. The ang 1 protein is expressed in testicular microvessels under basal conditions, and it is largely unaffected after hCG stimulation. Expression of ang 2 in microvessels, in contrast, is low under basal conditions and is up-regulated by hCG. Intratesticular injection of human recombinant ang 1 protein inhibits hCG-induced increase in vascular permeability. Injection of ang 2 in the testis increases endothelial cell proliferation and the volume of the interstitial space. We therefore suggest that ang 1 stabilizes testicular microvessels under basal conditions and that a shift in this balance caused by increased ang 2, together with increased VEGF-A, allows vascular leakage, high endothelial cell proliferation, and presumably, vascular growth after hormonal stimulation. PMID- 12773424 TI - Effects of pulsatile shear stress on nitric oxide production and endothelial cell nitric oxide synthase expression by ovine fetoplacental artery endothelial cells. AB - Placental blood flow, endothelial nitric oxide (NO) production, and endothelial cell nitric oxide synthase (eNOS) expression increase during pregnancy. Shear stress, the frictional force exerted on endothelial cells by blood flow, stimulates vessel dilation, endothelial NO production, and eNOS expression. In order to study the effects of pulsatile flow/shear stress, we adapted Cellco CELLMAX artificial capillary modules to study ovine fetoplacental artery endothelial (OFPAE) cells for NO production and eNOS expression. OFPAE cells were grown in the artificial capillary modules at 3 dynes/cm2. Confluent cells were then exposed to 10, 15, or 25 dynes/cm2 for up to 24 h. NO production by OFPAE cells exposed to pulsatile shear stress was inhibited to nondetectable levels by the NOS inhibitor l-NMMA and reversed by excess NOS substrate l-arginine. NO production and expression of eNOS mRNA and protein by OFPAE cells were elevated by shear stress in a graded fashion (P < 0.05). The rise in NO production with 25 dynes/cm2 shear stress (8-fold) was greater (P < 0.05) than that observed for eNOS protein (3.6-fold) or eNOS mRNA (1.5-fold). The acute shear stress-induced rise in NO production by OFPAE cells was via eNOS activation, whereas the prolonged NO rise occurred by elevations in both eNOS expression and enzyme activation. Thus, elevations of placental blood flow and physiologic shear stress may be partly responsible for the increases in placental arterial endothelial eNOS expression and NO production during pregnancy. PMID- 12773425 TI - Expression of vascular endothelial growth factor receptors during male germ cell differentiation in the mouse. AB - Overexpression of vascular endothelial growth factor (VEGF) in the testis of transgenic mice induces infertility, suggesting a potential role for VEGF in the process of spermatogenesis. Spermatogenesis occurs within the confines of the seminiferous tubules, and the seminiferous epithelium lining these tubules consists of Sertoli cells and germ cells in various stages of maturation. We investigated the source of VEGF and VEGF-target cells within the seminiferous tubules of the normal mouse testis. Sections of testes fixed in Bouin solution and embedded in paraffin were subjected to immunofluorescent staining with specific antibodies against VEGF, and its receptors, VEGFR-1 (Flt-1) and VEGFR-2 (Flk-1). Total RNA was extracted from isolated populations of Sertoli cells, type A spermatogonia, pachytene spermatocytes, and spermatids. Primer pairs specific for VEGF and its receptors were designed and reverse-transcriptase polymerase chain reaction (RT-PCR) was performed. Immunofluorescent studies indicated that VEGF is strongly expressed in the cytoplasm of Sertoli cells. VEGFR-1 and VEGFR-2 were not expressed by the Sertoli cell. In contrast, a differential expression of VEGF receptors was observed in germ cells. Although VEGFR-2 was expressed in the cytoplasm of type A spermatogonia, VEGFR-1 was expressed in the acrosomal region of spermatids and spermatozoa. Pachytene spermatocytes did not exhibit any staining. Further, we examined the transcription of VEGF and its receptors by RT PCR. VEGF was actively transcribed only in Sertoli cells. The transcription of VEGFR-2 was confined to type A spermatogonia. Interestingly, VEGFR-1 was transcribed both in pachytene spermatocytes and round spermatids. The mRNA expression of VEGFR-1 and VEGFR-2 in germ cells was inversely correlated during postnatal development of the mouse testis. Thus, VEGF may play a potential role in regulating the initial stages of the process of spermatogonial proliferation through VEGFR-2 and spermiogenesis through VEGFR-1. PMID- 12773426 TI - Nuclear transfer of synchronized african wild cat somatic cells into enucleated domestic cat oocytes. AB - The African wild cat is one of the smallest wild cats and its future is threatened by hybridization with domestic cats. Nuclear transfer, a valuable tool for retaining genetic variability, offers the possibility of species continuation rather than extinction. The aim of this study was to investigate the ability of somatic cell nuclei of the African wild cat (AWC) to dedifferentiate within domestic cat (DSH) cytoplasts and to support early development after nuclear transplantation. In experiment 1, distributions of AWC and DSH fibroblasts in each cell-cycle phase were assessed by flow cytometry using cells cultured to confluency and disaggregated with pronase, trypsin, or mechanical separation. Trypsin (89.0%) and pronase (93.0%) yielded higher proportions of AWC nuclei in the G0/G1 phase than mechanical separation (82.0%). In contrast, mechanical separation yielded higher percentages of DSH nuclei in the G0/G1 phase (86.6%) than pronase (79.7%) or trypsin (74.2%) treatments. In both species, pronase induced less DNA damage than trypsin. In experiment 2, the effects of serum starvation, culture to confluency, and exposure to roscovitine on the distribution of AWC and DSH fibroblasts in various phases of the cell cycle were determined. Flow cytometry analyses revealed that the dynamics of the cell cycle varied as culture conditions were modified. Specifically, a higher percentage of AWC and DSH nuclei were in the G0/G1 phase after cells were serum starved (83% vs. 96%) than were present in cycling cells (50% vs. 64%), after contact inhibition (61% vs. 88%), or after roscovitine (56% vs. 84%) treatment, respectively. In experiment 3, we evaluated the effects of cell synchronization and oocyte maturation (in vivo vs. in vitro) on the reconstruction and development of AWC-DSH- and DSH-DSH-cloned embryos. The method of cell synchronization did not affect the fusion and cleavage rate because only a slightly higher percentage of fused couplets cleaved when donor nuclei were synchronized by serum starvation (83.0%) than after roscovitine (80.0%) or contact-inhibition (80.0%). The fusion efficiency of in vivo and in vitro matured oocytes used as recipient cytoplasts of AWC donor nuclei (86.6% vs. 85.2%) was similar to the rates obtained with DSH donor nuclei, 83.7% vs. 73.0%, respectively. The only significant effect of source of donor nucleus (AWC vs. DSH) was on the rate of blastocyst formation in vitro. A higher percentage of the embryos derived from AWC nuclei developed to the blastocyst stage than did embryos produced from DSH nuclei, 24.2% vs. 3.3%, respectively (P < 0.05). In experiment 4, the effect of calcium in the fusion medium on induction of oocyte activation and development of AWC-DSH-cloned embryos was determined. The presence of calcium in the fusion medium induced a high incidence of cleavage of DSH oocytes (54.3%), while oocyte cleavage frequency was much lower in the absence of calcium (16.6%). The presence or absence of calcium in the fusion medium did not affect the fusion, cleavage, and blastocyst development of AWC-DSH-cloned embryos. In experiment 5, AWC-DSH-cloned embryos were transferred to the uteri of 11 synchronized domestic cat recipients on Day 6 or 7 after oocyte aspiration. Recipients were assessed by ultrasonography on Day 21 postovulation, but no pregnancies were observed. In the present study, after NT, AWC donor nuclei were able to dedifferentiate in DSH cytoplasts and support high rates of blastocyst development in vitro. Incomplete reprogramming of the differentiated nucleus may be a major constraint to the in vivo developmental potential of the embryos. PMID- 12773427 TI - A role for kit receptor signaling in Leydig cell steroidogenesis. AB - Kit and its ligand, Kitl, function in hematopoiesis, melanogenesis, and gametogenesis. In the testis, Kitl is expressed by Sertoli cells and Kit is expressed by spermatogonia and Leydig cells. Kit functions are mediated by receptor autophosphorylation and subsequent association with signaling molecules, including phosphoinositide (PI) 3-kinase. We previously characterized the reproductive consequences of blocking Kit-mediated PI 3-kinase activation in KitY(719F)/Kit(Y719F) knockin mutant male mice. Only gametogenesis was affected in these mice, and males are sterile because of a block in spermatogenesis during the spermatogonial stages. In the present study, we investigated effects of the Kit(Y719F) mutation on Leydig cell development and steroidogenic function. Although the seminiferous tubules in testes of mutant animals are depleted of germ cells, the testes contain normal numbers of Leydig cells and the Leydig cells in these animals appear to have undergone normal differentiation. Evaluation of steroidogenesis in mutant animals indicates that testosterone levels are not significantly reduced in the periphery but that LH levels are increased 5-fold, implying an impairment of steroidogenesis in the mutant animals. Therefore, a role for Kit signaling in steroidogenesis in Leydig cells was sought in vitro. Purified Leydig cells from C57Bl6/J male mice were incubated with Kitl, and testosterone production was measured. Kitl-stimulated testosterone production was 2-fold higher than that in untreated controls. The Kitl-mediated testosterone biosynthesis in Leydig cells is PI 3-kinase dependent. In vitro, Leydig cells from mutant mice were steroidogenically more competent in response to LH than were normal Leydig cells. In contrast, Kitl-mediated testosterone production in these cells was comparable to that in normal cells. Because LH levels in mutant males are elevated and LH is known to stimulate testosterone biosynthesis, we proposed a model in which serum testosterone levels are controlled by elevated LH secretion. Leydig cells of mutant males, unable to respond effectively to Kitl stimulation, initially produce lower levels of testosterone, reducing testosterone negative feedback on the hypothalamic pituitary axis. The consequent secretion of additional LH, under this hypothesis, causes a restoration of normal levels of serum testosterone. Kitl, acting via PI 3-kinase, is a paracrine regulator of Leydig cell steroidogenic function in vivo. PMID- 12773428 TI - Carrageenan formulation prevents macrophage trafficking from vagina: implications for microbicide development. AB - Considerable evidence suggests that human immunodeficiency virus (HIV)-infected macrophages and/or lymphocytes may mediate sexual transmission of HIV. We and others have previously demonstrated that when vitally stained donor mouse lymphocytes or macrophages are placed in the vaginas of mice, some of the stained cells can later be found in the iliac lymph nodes. The aim of this study was to assess the extent of mononuclear cell trafficking from the vagina and to test the possibility that carrageenan formulation, a sulfated polysaccharide formulation containing 3% PDR98-15 carrageenan (PC-515; FMC Biopolymer, Rockland, ME), a vaginal microbicide, would prevent vaginal transmigration of macrophages. When supravitally stained mouse macrophages and T cells were inoculated into the vagina of recipient mice, discrete numbers of donor cells migrated to the recipient iliac and inguinal lymph nodes and spleen. When recipient mice were preinoculated with the carrageenan formulation, the number of macrophages in lymph nodes and spleen was reduced by >90%. In contrast, a methylcellulose formulation, which is believed to be inactive, did not significantly reduce migration to the lymphoid organs. Our findings suggest that the carrageenan formulation blocks cell trafficking of macrophages from vagina and that blocking does not result from cytotoxicity. Blocking cell trafficking may help to prevent sexual transmission of HIV. PMID- 12773429 TI - Production of nuclear transfer-derived piglets using porcine fetal fibroblasts transfected with the enhanced green fluorescent protein. AB - A system for somatic cell nuclear transfer (SCNT) was developed and led to the successful production of GFP-transfected piglets. In experiment 1, two groups of SCNT couplets reconstructed with porcine fetal fibroblasts (PFF) and enucleated sow (S) or gilt oocytes (G): 1). received a simultaneous electrical fusion/activation (S-EFA or G-EFA groups), or 2). were electrically fused followed by activation with ionomycin (S-EFIA or G-EFIA groups), or 3). were subjected to electrical fusion and subsequent activation by ionomycin, followed by 6-dimethylaminopurine treatment (S-EFIAD or G-EFIAD groups). The frequency of blastocyst formation was significantly higher in S-EFA (26%) compared with that observed in the other experimental groups (P < 0.05), but not with S-EFIA (23%). Sow oocytes yielded significantly higher cleavage frequencies (68%-69%) and total cell numbers of blastocysts when compared with gilt oocytes, regardless of fusion/activation methods (P < 0.05). However, the ratio of inner cell mass (ICM)/total cells in G-EFA and S-EFA was significantly lower than in the other groups (P < 0.05). In experiment 2, SCNT couplets reconstructed with PFF cultured in the presence or absence of serum and enucleated sow oocytes were subjected to EFA. There were no effects of serum starvation on cell-cycle synchronization, developmental competence, total cell numbers, and ratio of ICM/total cells. In experiment 3, SCNT couplets reconstructed with PFF transfected with an enhanced green fluorescence protein (EGFP) gene using FuGENE-6 and enucleated sow oocytes were subjected to EFA and cultured for 7 days. Expression frequencies of GFP gene during development were 100%, 78%, 72%, 71%, and 70% in fused, two-cell, four to eight cells, morulae, and blastocysts, respectively. In experiment 4, SCNT embryos derived from different recipient cytoplasts (sows or gilts) and donor karyoplasts (PFF or GFP-transfected) were subjected to EFA and transferred to the oviducts of surrogates. The pregnancy rates in SCNT embryos derived from sow oocytes (66%-69%) were higher than those with gilt oocytes (23%-27%) regardless of donor cell types. One live offspring from GFP-SCNT embryos and two from PFF SCNT embryos were delivered. Microsatellite analysis confirmed that the clones were genetically identical to the donor cells and polymerase chain reaction (PCR) from genomic DNA of cloned piglets and subsequent southern blot analysis confirmed the integration of EGFP gene into chromosomes. PMID- 12773430 TI - Increased messenger RNA for allograft inflammatory factor-1, LERK-5, and a novel gene in 17.5-day relative to 15.5-day bovine embryos. AB - Considerable embryonic loss occurs between Gestation Days 15 and 18 in cattle when critical cellular and molecular events occur, including maternal recognition of pregnancy. To gain insight into these events, mRNA differential display analysis was used to identify eight unique cDNA fragments present in greater abundance in 17.5-day than in 15.5-day bovine embryos. Four cDNA fragments, confirmed to be upregulated in 17.5-day embryos using Northern analysis, were cloned and sequenced. Three cDNA fragments shared sequence identities with known homologs: human allograft inflammatory factor-1 (AIF-1), human LERK-5, and bovine interferon-tau. One novel cDNA fragment did not share sequence identity to previously reported genes, except for a similar DNA sequence in the human genome. AIF-1 mRNA was present in developing placenta through Gestation Day 36, and abundant levels were observed in adult bovine spleen and lung. The novel gene, which we have named periattachment factor (PAF), was not detected in adult tissues using Northern analysis or in conceptuses between Days 30 and 36 of pregnancy. Additional sequence information for bPAF was obtained from a cDNA library constructed from a 25-day bovine embryo. The protein corresponding to the open reading frame has four protein kinase C phosphorylation sites, two casein kinase II phosphorylation sites, a nuclear targeting sequence, but no obvious DNA or RNA binding motifs. Abundant expression of this gene during a narrow but critical window of embryonic development makes it worthy of further study. PMID- 12773432 TI - Effect of mouse uterine stromal cells on epithelial cell transepithelial resistance (TER) and TNFalpha and TGFbeta release in culture. AB - Recognizing that uterine stromal cells regulate several uterine epithelial cell function(s), the current study was undertaken to more fully define cell-cell communication in the uterus and to examine the role of uterine stromal cells in regulating epithelial cell monolayer integrity and cytokine release. Uterine epithelial and stromal cells from adult intact mice were isolated and cultured separately on cell culture inserts and/or in culture plates. Epithelial cells, which reach confluence as indicated by high transepithelial resistance (TER > 1000 ohms/well), preferentially release transforming growth factor-beta (TGFbeta) into the basolateral chamber ( approximately 70% > apical) and tumor necrosis factor-alpha (TNFalpha) into the apical compartment ( approximately 30% > basolateral). When epithelial cells on cell culture inserts were transferred to plates containing stromal cells, coculture for 24-48 h increased epithelial cell TER ( approximately 70% higher than control) and decreased TNFalpha release into both the apical and basolateral chambers ( approximately 30%-50%). In contrast, TGFbeta release was not affected by the presence of stromal cells. In other studies, the effects of stromal cells on epithelial cell TER and TNFalpha release persisted for 5-7 days following the removal of stromal cells and were also seen in coculture studies in which conditioned stromal media (CSM) was placed in the basolateral chamber. These studies indicate that uterine stromal cells produce a soluble factor(s) that regulates epithelial cell TER and release of TNFalpha without effecting TGFbeta release. These results suggest that uterine stromal cells communicate with epithelial cells via a soluble factor(s) to maintain uterine integrity and epithelial secretory function. PMID- 12773431 TI - Determination of steroidogenic potential of ovarian cells of the brushtail possum (Trichosurus vulpecula). AB - The ovary of the brushtail possum (Trichosurus vulpecula) secretes steroids; however, little is known about the identity of the steroidogenic cells in the ovary. The aim of the present study was to determine the identity of the ovarian cell types expressing mRNAs encoding proteins important for steroidogenesis and determine at what stage of follicular development they are expressed. The genes examined were those for steroidogenic factor-1 (SF-1), steroidogenic acute regulatory protein (StAR), cytochrome p450 side chain cleavage (P450scc), 3beta hydroxysteroid dehydrogenase/Delta5,Delta4 isomerase (3betaHSD), cytochrome p45017alphahydroxylase (p45017alphaOH), and p450 aromatase (p450arom). None of the genes examined were expressed in oocytes at any stage of follicular development. SF-1 was expressed in granulosa cells from the type 2 or the primary stage of development and thereafter to the preovulatory stage. In addition, the theca interna of small and medium-size antral but not preovulatory follicles and the interstitial glands and corpora lutea expressed SF-1 mRNA. Granulosa cells of preantral and small to medium-size antral follicles were not capable of synthesizing steroids from cholesterol because they did not contain p450scc mRNA. However, granulosa cells of many of the small to medium-size antral follicles expressed p450arom and 3betaHSD mRNA. The interstitial glands, theca interna, and corpus luteum expressed StAR, p450scc, 3betaHSD, and p45017alphaOH mRNA, suggesting that these tissues are capable of synthesizing progestins and androgens. The corpus luteum expressed p450arom, indicating that this tissue also has the potential to secrete estrogens in this species. PMID- 12773433 TI - Effects of endotoxin and macrophage-related cytokines on the contractile activity of the gravid murine uterus. AB - Immune cell trafficking and activity are implicated in the parturition process, but little is known about the role of macrophages in control of uterine contractility at term. In the present study, we tested the hypothesis that endotoxin (lipopolysaccharide [LPS]) enhances uterine contractile activity through a mechanism that involves activation of resident macrophages. Various uterotonins and anti-inflammatory mediators were added to a standard muscle bath preparation that contained strips of uterus from Day 15 pregnant C3H/HeN mice. Spontaneous and agonist-induced contractile activity was enhanced following LPS treatment. LPS increased amplitude but not frequency of contractions. Addition of anti-inflammatory cytokines, interleukin 10 or transforming growth factor beta, to suppress macrophage activation did not block LPS-induced increases in contractility. By contrast, indomethacin given to block prostaglandin production suppressed the LPS-induced increase in amplitude of contractions. These findings suggest that an inflammatory response, possibly mediated by activation of macrophages and prostaglandins, participates in the regulation of amplitude but not frequency of contractile activity by the murine uterus before onset of parturition. PMID- 12773434 TI - Effect of growth hormone (GH) on in vitro nuclear and cytoplasmic oocyte maturation, cumulus expansion, hyaluronan synthases, and connexins 32 and 43 expression, and GH receptor messenger RNA expression in equine and porcine species. AB - The aim of this study was to investigate the role of growth hormone (GH) on in vitro cumulus expansion and oocyte maturation in equine and porcine cumulus oocyte complexes (COCs), and to approach its way of action. Equine COCs were cultured in a control medium (TCM199, 5 mg/ml BSA, 1 microg/ml estradiol, and antibiotics) supplemented with either 0.5 microg/ml equine GH or 5 microg/ml equine LH. Porcine COCs were cultured in a basal medium (TCM199 with 570 microM cysteamine) supplemented with 0, 0.1, 0.5, or 1 microg/ml porcine GH or in a control medium (basal medium with 10 ng/ml epidermal growth factor and 400 ng/ml FSH) supplemented with 0 or 0.5 microg/ml porcine GH. After culture, cumulus expansion and nuclear stage were assessed. The cytoplasmic maturation of porcine oocytes was evaluated by in vitro fertilization and development for 7 days. The modifications of the expression of proteins implicated in cumulus expansion were analyzed in equine COCs by SDS-PAGE with antibodies against connexins 32 and 43 and hyaluronan synthases (Has) 1, 2, and 3. The expression of GH receptor mRNA was studied in oocytes and cumulus cells of the two species using reverse transcription-polymerase chain reaction with specific primers. The addition of GH in maturation medium increased cumulus expansion in equine but not porcine COCs. It improved nuclear maturation in equine and porcine, but had no effect on porcine fertilization and embryo development. The GH receptor mRNA was detected in equine and porcine oocytes and cumulus cells. GH did not influence the expression of Has 1, Has 3, and connexin 43 in equine cumulus cells. PMID- 12773436 TI - Early ovarian ageing: a hypothesis. Detection and clinical relevance. AB - An important scientific breakthrough of the 20th century, IVF helped open new horizons in medicine, such as pre-implantation genetic diagnosis and embryonic stem-cell therapy. A further contribution of assisted reproduction technology has been the better understanding of reproductive ageing. Data from IVF cycles suggest that there is a fixed time-interval between accelerated decline of fertility and the menopause. This leads to the hypothesis that a significant proportion of asymptomatic women in the early thirties may be at risk of early onset of subfertility. IVF provides a model for the development of ovarian reserve tests, some of which appear promising as potential screening tools for early ovarian ageing in the general population. PMID- 12773435 TI - Homocysteine induces trophoblast cell death with apoptotic features. AB - Hyperhomocysteinemia has been suggested as a possible risk factor in women suffering from habitual abortions, placental abruption or infarcts, preeclampsia, and/or intrauterine growth retardation. However, little is known about the pathogenic mechanisms underlying the action of homocysteine. The present study investigated the in vitro ability of homocysteine to affect trophoblast gonadotropin secretion and to induce cell death. In primary human trophoblast cells, homocysteine treatment (20 micromol/L) resulted in cellular flattening and enlargement, extension of pseudopodia, and cellular vacuolization. Cellular detachment, apoptosis, and necrosis were favored. With in situ nick end labeling, we investigated DNA degradation, and we used M30 CytoDEATH to selectively stain the cytoplasm of apoptotic cells. Cytochrome c release from mitochondria to the cytosol and DNA cleavage in agarose gel have been investigated. Homocysteine, but not cysteine, induced trophoblast apoptosis and significantly reduced human chorionic gonadotropin secretion. These findings suggest that trophoblast cell death might represent a pathogenic mechanism by which homocysteine may cause pregnancy complications related to placental diseases. PMID- 12773437 TI - Prediction of ovarian hyperstimulation syndrome (OHSS). Estradiol level has an important role in the prediction of OHSS. AB - The cascade of events that leads to the development of ovarian hyperstimulation syndrome (OHSS) is almost always accompanied by elevated estradiol (E(2)) levels. The role of estrogen level in OHSS has not been confirmed; however, it was shown repeatedly in the literature that elevated levels of E(2) constitute a risk for OHSS. Monitoring E(2) was found to be effective in reducing the incidence of OHSS. It was also reported that reducing the E(2) level by coasting helps to prevent OHSS. It is believed that irrespective of the debatable role of estrogen in the pathogenesis of OHSS, there is a general agreement that E(2) assay is an important marker to detect the majority of patients at risk of OHSS. PMID- 12773438 TI - Recombinant FSH in alternative doses or versus urinary gonadotrophins for ovulation induction in subfertility associated with polycystic ovary syndrome: a systematic review based on a Cochrane review. AB - This paper is based on a Cochrane review published in The Cochrane Library, issue 2, 2001 (see www.CochraneLibrary. net for information) with permission from The Cochrane Collaboration and Update Software. Cochrane reviews are regularly updated as new evidence emerges and in response to comments and criticisms, and The Cochrane Library should be consulted for the most recent version of the review. This systematic review was performed to study the efficacy and safety of recombinant FSH (rFSH) versus urinary FSH (uFSH) and to compare different dose regimens of rFSH for ovulation induction in women with clomiphene-resistant polycystic ovary syndrome (PCOS). Six randomized controlled trials were included in the review, according to the principles of the Cochrane Menstrual Disorders and Subfertility Group. Three trials compared rFSH with uFSH and three trials compared two different treatment regimens of rFSH. Participants were women with clomiphene citrate-resistant PCOS. Main outcome measures were ovulation, clinical pregnancy, miscarriage, multiple pregnancy, ovarian stimulation syndrome (OSS), total gonadotrophin dose used and total duration of stimulation. Summary statistics were expressed as odds ratios. Data from the trials comparing rFSH and uFSH could be pooled. There was no evidence of a difference between rFSH and uFSH in any of the outcomes. Data from the trials comparing different dose regimens of rFSH could not be combined, and for each comparison there was insufficient evidence of a difference. More randomized clinical trials with sufficient power are necessary to estimate the difference, if one exists, between rFSH and uFSH and between different dose regimens of rFSH. PMID- 12773439 TI - A detailed cytogenetic analysis of large numbers of fresh and frozen-thawed human sperm after ICSI into mouse oocytes. AB - BACKGROUND: Since information about chromosome aberrations in micro-manipulated sperm is still inadequate, cytogenetic analysis was performed on large numbers of fresh and frozen-thawed (FT) human sperm after injection into mouse oocytes. The effects of the ICSI procedure on oocytes are also discussed based on analysis of the mouse chromosome complements. METHODS: After the injection of fresh and FT human sperm into mouse oocytes, chromosomes of the hybrid oocytes were analysed at first cleavage metaphase. RESULTS: Incidences of the hybrid oocytes at the first cleavage metaphase were significantly different between fresh (71.5%) and FT sperm groups (80.1%) (P < 0.05). The chromosome analysis of 477 fresh and 141 FT sperm showed no difference in the incidences of aneuploidy (1.6/0.7%), structural aberrations (8.8/7.8%) or diploidy (0.0/0.0%) between these categories. The cytogenetic result did not differ from our previous result using IVF between human sperm and hamster oocytes. In an additional cytogenetic study on 615 mouse chromosome complements, the incidence of diploidy (5.4%) was significantly higher than those (0.3-2.8%) in the previous mouse cytogenetic studies, and the hybrid oocytes with no mouse chromosomes (2.0%) existed. CONCLUSIONS: This result suggests that the ICSI procedure induces no sperm chromosome aberrations but increases numerical aberrations in oocyte chromosome complements. PMID- 12773440 TI - Density and distribution of primordial follicles in single pieces of cortex from 21 patients and in individual pieces of cortex from three entire human ovaries. AB - BACKGROUND: At the time of cryopreservation of ovarian tissue for fertility preservation a small biopsy of ovarian cortex is usually taken for histological evaluation of the follicular reserve. The purpose of this study was to evaluate the distribution and density of primordial follicles in single pieces of cortex from individual patients and in pieces of cortex comprising entire ovaries, all prepared for cryopreservation. METHODS: Cortical biopsies from 21 patients and the whole cortex of one ovary were evaluated histologically prior to cryopreservation. In addition, the cortex of two whole ovaries was cryopreserved before histological evaluation. The volume of each cortical fragment was measured, all follicles counted and the follicular density calculated. RESULTS: In individual pieces of cortex follicular density showed a significant inverse linear correlation with age. The follicular density per cortical fragment prepared from each of the three entire ovaries varied from 1.8 to 166, 0.007 to 140 and 0.04 to 4.48 follicles/mm(3) cortical tissue. CONCLUSION: The density of primordial follicles varied more than two orders of magnitude in cortical fragments from each of the three ovaries. Primordial follicles were very unevenly distributed throughout the cortex of these ovaries, although a significant linear correlation between age and follicular density was found. PMID- 12773441 TI - Transplantation of intact rat gonads using vascular anastomosis: effects of cryopreservation, ischaemia and genotype. AB - BACKGROUND: A limited store of ovarian follicles is present at birth and its progressive decline during ageing is hastened by alkylating agents and ionizing radiation during treatment for cancer or autoimmune disease. Oligo- or azoospermia can arise for similar reasons in men. There is some experimental evidence showing that targeted gene deletion or drugs to produce hypogonadotrophism can protect germ cells from wastage. Another strategy for conserving fertility is to cryopreserve ovarian or testicular tissue for subsequent transplantation. To maximize gonadal function, it is desirable to preserve whole gonads for transplantation using vascular anastomosis. METHODS AND RESULTS: We investigated this strategy in the rat model. All freshly isotransplanted ovaries (n = 8) survived and resumed follicle growth and secretion and, although ischaemia for 24 h at 4 degrees C did not disrupt ovarian function, the organs had fewer follicles. Four out of seven (57%) cryopreserved transplants survived for > or =60 days, were ovulatory and one pregnancy was established, but the ovarian reserve was compromised by fewer follicles. Ovarian allotransplants were vigorously rejected, even with moderate immunosuppression using cyclosporin A. On the other hand, only three out of seven (42%) fresh testicular isotransplants had active spermatogenesis, and none of the cryopreserved testes was functional. CONCLUSIONS: The effects of gonadectomy in rats can be reversed by isotransplants, but the results are more successful with ovaries than testes, and allotransplants were never successful. Intact cryopreserved ovaries can be restored to function after transplantation with vascular anastomoses. PMID- 12773442 TI - Cytokine production by peripheral blood monocytes during the normal human ovulatory menstrual cycle. AB - BACKGROUND: There is considerable evidence that hormone-driven changes resembling an inflammatory response occur in the vascular compartment during the menstrual cycle, and peripheral blood monocytes may be central in the process. We investigated whether there is a cyclical change in intrinsic production of pro inflammatory cytokines by monocytes in the ovulatory menstrual cycle, and whether there is a circulating factor that influences the pattern of cytokine production in a cyclical manner. METHODS: Monocytes were purified by density-gradient centrifugation followed by countercurrent centrifugal elutriation, from the blood of normal women (n = 10) pre- and post-ovulation. Monocytes were cultured under basal conditions with bacterial lipopolysaccharide (LPS), and to determine the effects of circulating factors, incubations were also conducted in the presence of autologous serum. Concentrations of interleukin (IL)-1alpha, IL-1beta, IL-6, tumour necrosis factor (TNF)-alpha and IL-1 receptor antagonist (IL-1Ra) were measured by sandwich ELISA. RESULTS: The majority of IL-1alpha and IL-1beta was cell associated, while the other cytokines were almost entirely secreted. Basal levels of IL-1alpha, IL-1beta, and TNF-alpha were significantly increased following ovulation, while there was no significant change in levels of secretion of IL-6 or IL-1Ra. These effects were present in unstimulated cells, suggesting prior activation in vivo. Cytokine production was increased in response to LPS; however, there was no consistent effect of autologous serum. CONCLUSIONS: Intrinsic production of pro-inflammatory cytokines by monocytes is increased following ovulation. PMID- 12773443 TI - Leukocyte behaviour and permeability in the rat mesenteric microcirculation following induction of ovulation. AB - BACKGROUND: Ovarian hyperstimulation syndrome (OHSS), a highly dangerous and incompletely understood complication of ovulation induction with exogenous gonadotrophins, can include haemoconcentration, hypovolaemia, hypotension, acute renal insufficiency, thromboembolism and ultimately death. Using intravital microscopy, we examined microvascular permeability and leukocyte-endothelial cell interactions in the rat mesenteric microcirculation associated with induction of ovulation. METHODS: In female rats treated with hMG and hCG, mesenteric venules were observed by intravital microscopy assisted by a video imager. Erythrocyte velocity was monitored, and rolling and adhesion of leukocytes were studied by transmission video images. Transvascular leakage of fluorescein isothiocyanate labelled albumin was assessed by epi-illumination. RESULTS: Administration of hMG and hCG significantly increased vascular protein leakage within a few hours, and also reduced rolling velocities of leukocytes in venules and increased numbers of leukocytes adherent to endothelium at 16 h following hCG injection. The administration of antibodies against intracellular adhesion molecule (ICAM)-1 inhibited these reactions. CONCLUSION: By induction of ovulation, vascular permeability is increased not only at the surface of the ovary but also in the mesentery. Alteration of leukocyte behaviour in the microcirculation through mechanisms involving ICAM-1 is one likely cause of the protein leakage. PMID- 12773444 TI - Immunocytochemical assessment of mitotic activity with an antibody to phosphorylated histone H3 in the macaque and human endometrium. AB - BACKGROUND: Determination of the mitotic index in sections of endometrium stained with haematoxylin and eosin (H&E) is difficult and time-consuming. We assessed the value of two mitotic markers as immunocytochemical reagents for measuring mitotic rates in endometrium. METHODS: Mitotic protein monoclonal antibody 2 (MPM 2) and anti-phosphorylated histone H3 (Phospho H3) were applied to paraffin sections of rhesus macaque and human endometrium. RESULTS: In estrogen-treated macaque endometrium the mean +/- SEM mitotic indices were: H&E 1.5 +/- 0.25%, Phospho H3 antibody 1.02 +/- 0.23% and MPM-2 antibody 0.69 +/- 0.17%; these were not statistically significantly different, but the Phospho H3 antibody gave a stronger and cleaner signal than the MPM-2 antibody. Comparisons were made between a computer-determined Phospho H3 index, the H&E-determined mitotic index and the Ki-67 index in samples of human endometrium across the cycle. All revealed that the highest proliferative rate occurred during the follicular phase, but the Phospho H3 and the mitotic indices were more highly correlated (R(2) = 0.89, P < 0.001) than the Ki-67 and mitotic indices (R(2) = 0.74, P < 0.05). CONCLUSIONS: The exceptionally high contrast staining and the excellent correlation between the Phospho H3 and mitotic indices validates the specificity of the Phospho H3 antibody as a new tool for the assessment of endometrial mitotic activity. PMID- 12773445 TI - A prospective, randomized, controlled trial comparing highly purified hMG with recombinant FSH in women undergoing ICSI: ovarian response and clinical outcomes. AB - BACKGROUND: To assess the clinical profile and efficacy in assisted reproductive treatment of a new human-derived highly purified (HP) menotropin, we compared HP hMG and recombinant (r) FSHalpha use in ICSI within a prospective, randomized, controlled study. METHODS: 100 infertile women were treated with HP hMG (50 patients) or rFSHalpha (50 patients). All patients received the same daily gonadotrophin dose (150 IU) following GnRH agonist suppression (long regimen) until more than three follicles >17 mm and estradiol (E(2)) levels >600 pg/ml were reached. Patients were monitored with daily LH, FSH, hCG, estradiol (E(2)), progesterone, and testosterone measurements; and alternate day pelvic ultrasound. RESULTS: Treatment duration (11.1 +/- 0.4 versus 12.9 +/- 0.5 days, P < 0.05) and gonadotrophin dose (22.4 +/- 1.0 versus 27.0 +/- 1.5 ampoules, P < 0.05) were lower in the HP hMG group. Conversely, peak pre-ovulatory E(2) (1342 +/- 127 versus 933 +/- 109 pg/ml, P < 0.005); and area under the curve of E(2) (3491 +/- 350 versus 2602 +/- 349 pg/ml.day, P < 0.05), immunoreactive serum FSH (65.9 +/- 2.1 versus 48.8 +/- 1.8 IU/l.day, P < 0.001). and hCG (1.7 +/- 0.3 versus 0.0 +/- 0.0 IU/l/day, P < 0.001) during treatment were higher in the HP hMG group. Cycle cancellation rates, transferred embryo number, pregnancy rates per started cycle (30 versus 28%) and per embryo transfer (35 versus 35%) and miscarriage rates (6 versus 6%) were not significantly different. CONCLUSIONS: HP hMG treatment was associated with: (i) a more efficient patient response, as reflected by reduced treatment duration and gonadotrophin requirements; (ii) increased serum levels of hCG, E(2), and immunoreactive FSH during treatment; (iii) an ICSI outcome indistinguishable from rFSHalpha. PMID- 12773447 TI - Inverse correlation between peritoneal fluid leptin concentrations and the extent of endometriosis. AB - BACKGROUND: The role of leptin in reproductive processes has received increasing attention. Because leptin has intrinsic angiogenic properties, may be induced by inflammatory cytokines and induces matrix metalloproteinases, we examined peritoneal fluid (PF) leptin concentrations in women with endometriosis. METHODS: PF samples were collected from 60 women undergoing laparoscopy for endometriosis, and 18 controls undergoing tubal sterilization. Fifty of the women with endometriosis had received no prior hormonal treatment, while 10 with moderate- severe endometriosis were using GnRH agonists. RESULTS: Women with untreated endometriosis had significantly higher (mean +/- SD) PF leptin levels (34.9 +/- 7.9 ng/ml) than controls (17.9 +/- 4.1 ng/ml; P < 0.001). However, PF leptin levels were inversely correlated with the stage of disease (r = -0.62; P < 0.001). Nevertheless, women with stage III-IV endometriosis maintained significantly higher PF leptin levels (26.3 +/- 4.8 ng/ml; P < 0.001) than controls. Although PF leptin levels were significantly higher in the secretory versus proliferative phase of the menstrual cycle, they remained higher in both phases in women with untreated endometriosis. PF leptin levels in women on GnRH agonists were similar to controls. CONCLUSIONS: PF leptin levels are elevated in women with endometriosis, but inversely correlated with extent of disease. These findings suggest a potential role for leptin in the pathogenesis of peritoneal endometriosis. PMID- 12773446 TI - An open, randomized single-centre study to compare the efficacy and convenience of follitropin beta administered by a pen device with follitropin alpha administered by a conventional syringe in women undergoing ovarian stimulation for IVF/ICSI. AB - BACKGROUND: A pen device, similar to an insulin pen, has been recently marketed for the administration of follitropin beta in cartridges. A randomized controlled trial was performed to compare the efficacy and convenience of this pen device delivering follitropin beta with a conventional syringe delivering follitropin alpha. METHODS: A total of 200 patients needing IVF/ICSI treatment and willing to self-inject were enrolled in the study. All subjects had ovarian stimulation according to a long protocol and were randomized to the pen or the conventional syringe group during down-regulation by means of a computer-generated randomization list using random numbers. Patients were asked to fill in a daily local tolerance book after each injection. On the day of hCG the patients scored a Visual Analogue Scale (VAS) for pain and convenience. RESULTS: The average duration, total dose of recombinant FSH and number of cumulus oocyte complexes retrieved were 10.8/12.0 days (P = 0.001), 1880/2226 IU (P < 0.001) and 15.2/13.1 respectively in the pen device and conventional syringe groups; the presence of pain after the daily injection was significantly higher in the conventional syringe group (P = 0.027); the visual analogue scale score was similar for pain but significantly more convenient for the pen device (P < 0.001). The live birth rate per embryo transfer was 32.9 and 34.4% respectively in the pen device and conventional syringe groups. CONCLUSIONS: Self-injection with the pen device is safe and easy, more convenient and less painful for the patient, requires less FSH and shortens the treatment duration. PMID- 12773448 TI - Selective effects of pioglitazone on insulin and androgen abnormalities in normo- and hyperinsulinaemic obese patients with polycystic ovary syndrome. AB - BACKGROUND: To investigate the effectiveness and safety of pioglitazone (45 mg/day) on clinical and endocrine-metabolic features of polycystic ovary syndrome (PCOS), we studied 18 obese PCOS patients, classified as normoinsulinaemic (N PCOS, n = 6) and hyperinsulinaemic (H-PCOS, n = 12) according to their insulin secretion. METHODS: Evaluation of clinical signs, hormonal and lipid profile assays, oral glucose tolerance tests and euglycaemic hyperinsulinaemic clamps were performed at baseline and after 2 (visit 2), 4 (visit 3) and 6 (visit 4) months of treatment. RESULTS: Body weight, body fat distribution and blood pressure remained stable throughout the treatment; hirsutism and acne significantly improved (P < 0.001 for visits 3 and 4 versus baseline) in both groups. A restoration of menstrual cyclicity was observed at visit 4 in 83% (P < 0.001) of H-PCOS and in 33% of N-PCOS. A decrease in LH, LH/FSH ratio, androstenedione and 17-hydroxy-progesterone was observed in both groups, attaining statistical significance in H-PCOS. A significant amelioration of insulin secretion, sensitivity and clearance was obtained in H-PCOS. A trend towards improvement was observed in lipid assessment of both groups. Therapy was well-tolerated. CONCLUSIONS: Present data suggest that there is a selective effect, partially independent of insulin secretion, of pioglitazone on the clinical and hormonal disturbances of PCOS. PMID- 12773449 TI - Dual renin-angiotensin blockade therapy in patients at high risk of early ovarian hyperstimulation syndrome receiving IVF and elective embryo cryopreservation: a case series. AB - BACKGROUND: Ovarian hyperstimulation syndrome (OHSS) is an important and dangerous aspect of assisted reproduction techniques. Although elective cryopreservation of all embryos can prevent pregnancy-induced late OHSS, it cannot prevent early OHSS, which is induced by hCG administration. METHODS: We undertook this trial to assess the efficacy with which the combined oral administration of angiotensin-converting enzyme inhibitor (ACEI) and angiotensin II receptor blocker (ARB) could prevent early OHSS in IVF patients at very high risk for this syndrome. Four women, who had serum estradiol concentration > or =8000 pg/ml on the day of hCG injection, were treated with the combination of the ACEI alacepril and the ARB candesartan cilexetil for 8 days starting the day after oocyte retrieval. Embryos were cryopreserved and embryo transfer was postponed until later cycles. RESULTS: Despite the extremely enlarged ovaries, no ascites was accumulated in any of the cases. Haematocrit (34.1 +/- 1.0) and serum albumin concentration (4.1 +/- 0.2 g/dl) were normal throughout the treatment period. These patients showed elevated plasma renin and angiotensin II concentration before the treatment. CONCLUSIONS: The dual renin-angiotensin blockade therapy used here would be worth exploring further in a study with more patients and a prospective, randomized design. PMID- 12773450 TI - Hysteroscopic sterilization using a micro-insert device: results of a multicentre Phase II study. AB - BACKGROUND: Unlike laparoscopic surgery for interval tubal sterilization, a hysteroscopic approach obviates surgical incision and requires only local anaesthesia or intravenous sedation. The safety, tolerability and efficacy of an hysteroscopically placed micro-insert device was evaluated. METHODS: A cohort of 227 previously fertile women participated in this prospective international multicentre trial. Micro-inserts were placed bilaterally into the proximal Fallopian tube lumens under hysteroscopic visualization in outpatient procedures. RESULTS: Successful bilateral micro-insert placement was achieved in 88% of women. The majority of women reported that intraprocedural pain was less than or equal to that expected, and 90% rated tolerance of the device placement procedure as good to excellent. Most women could be discharged in an ambulatory state within 1-2 h. Adverse events occurred in 7% of the women, but none was serious. Correct device placement was confirmed in 97% of cases at 3 months. Over 24 months follow-up, 98% of study participants rated their tolerance of the micro insert as very good to excellent. After 6015 woman-months of exposure to intercourse, no pregnancies have been recorded. CONCLUSIONS: Hysteroscopic sterilization resulted in rapid patient recovery without unacceptable post procedure pain, as well as high long-term patient tolerability, satisfaction and effective permanent contraception. PMID- 12773451 TI - Management of a perforated levonorgestrel-medicated intrauterine device--a pharmacokinetic study: case report. AB - Intrauterine contraception is a widely used, highly effective method of birth control. Uterine perforation is a serious albeit rare complication with the use of an intrauterine device (IUD). Although uterine perforation by the levonorgestrel-releasing intrauterine system (LNG-IUS) has already been described, no plasma LNG concentrations in this setting were reported. Neither has the management of LNG-IUS been commented on to date. Two months after insertion of an LNG-IUS into a 33-year-old woman, it was noted to be in the peritoneal cavity. Laparoscopy for IUD removal was conducted 5 months after insertion. LNG and sex hormone-binding globulin plasma concentrations were measured prior to and following the laparoscopic removal of the IUD. Intra peritoneal dislocated LNG-IUS resulted in plasma LNG levels 10 times higher (4.7 nmol/l) than the plasma level of LNG observed with LNG-IUS placed in utero. This high plasma LNG level suppresses ovulation. Therefore a misplaced LNG-IUS should be removed when pregnancy is desired. PMID- 12773452 TI - Morbidity in a Danish national cohort of 472 IVF/ICSI twins, 1132 non-IVF/ICSI twins and 634 IVF/ICSI singletons: health-related and social implications for the children and their families. AB - BACKGROUND: There is a lack of knowledge on child health as well as family well being in IVF/ICSI twins. METHODS: These data originated from questionnaires completed by mothers taking part in a national cohort study of twin and singleton births occurring in Denmark in 1997. The overall response rate was 83%. The three cohorts consisted of all IVF/ICSI twin children (n = 472), all IVF/ICSI singletons (n = 634) and all non-IVF/ICSI twin children (n = 1132) born in Denmark in 1997. RESULTS: No major differences in physical health were observed between IVF/ICSI twins and non-IVF/ICSI twins. Compared with IVF/ICSI singletons, more IVF/ICSI twins were admitted to a neonatal intensive care unit (NICU) (P < 0.01) and more had surgical interventions (P = 0.03) and special needs (P = 0.02), moreover they had poorer speech development (P < 0.01). Correspondingly, IVF/ICSI twin mothers rated their infant's general health poorer than IVF/ICSI singleton mothers did. All discrepancies between IVF/ICSI twins and singletons disappeared after stratification for birthweight except for NICU admissions and speech development. Multiple logistic regression analyses showed that both IVF/ICSI and non-IVF/ICSI twin parents experienced more marital stress [odds ratio (OR) 2.9, 95% CI 2.2-3.8] and that twins had more impact on the mother's life (OR 1.7, 95% CI 1.2-2.4) compared with singletons. Nevertheless, the only predictor of low divorce/separation risk was IVF/ICSI treatment. CONCLUSION: Our study indicates that physical health of IVF/ICSI twins is comparable with that of non-IVF/ICSI twins. However, physical health of IVF/ICSI twins is poorer and the implications for the families stronger compared with IVF/ICSI singletons. PMID- 12773454 TI - Human oocyte cryopreservation as an adjunct to IVF-embryo transfer cycles. AB - BACKGROUND: The purpose of this work was to develop methods for successful cryopreservation of human oocytes. METHODS: Two cryopreservation procedures were used. Method 1 involved use of 1.5 mol/l propanediol (PrOH)-0.1 mol/l sucrose with medium containing sodium (Na) as cryoprotectant medium, seeding at -7 degrees C, and stepwise dilution of cryoprotectant post-thaw. Method 2 used Na depleted media with 1.5 mol/l PrOH-0.2 mol/l sucrose for freezing, seeding at -6 degrees C, and use of high sucrose (0.5 and 0.2 mol/l) for cryoprotectant removal. RESULTS: The first method was used in seven patients, and gave poor (12.3%) survival results and no pregnancies. The second method was used in 15 patients (16 cycles), and yielded good survival and fertilization rates (74.4 and 59% respectively), with four pregnancies and five healthy infants born to 11 women receiving an embryo transfer. CONCLUSIONS: Using Na-depleted media along with other alterations in freezing and thawing procedures, human oocyte cryopreservation can provide excellent survival and pregnancy rates. PMID- 12773453 TI - Assisted reproduction techniques for HIV serodiscordant couples: 18 months of experience. AB - BACKGROUND: Assisted reproduction techniques can minimize the risk of infection and treat possible sterility associated with serodiscordant couples. METHODS: We assessed the efficacy of these techniques in 57 couples in which at least one partner had human immunodeficiency virus (HIV-1) infection that was currently under control (47 men and 10 women). The semen of seropositive men was prepared and tested for viruses. Assisted reproduction techniques included intrauterine insemination (IUI), IVF and especially ICSI, with ovarian stimulation that used a long agonist protocol and recombinant FSH. Embryos were transferred on day 3 after oocyte retrieval. RESULTS: For couples with seropositive men, five IUI and 49 IVF or ICSI attempts were perfomed, whilst for seropositive women these numbers were three IUI and 12 IVF or ICSI. No pregnancy occurred following the eight IUI trials. Seroconversion was not observed in any partners of seropositive men. Efficacy of treatment for these couples with ICSI was good, the clinical pregnancy rate per embryo transfer was 48.8%. The results for seropositive women were disappointing, with a clinical pregnancy rate per embryo transfer of 9.1%. Fourteen babies from 47 treated couples have so far been born and no pregnancies from IUI. CONCLUSIONS: Assisted reproduction techniques and particularly ICSI provide HIV-1-seropositive men with a safe and highly effective means of fathering children. These techniques may be less effective for seropositive women. PMID- 12773455 TI - The administration of the GnRH antagonist, cetrorelix, to ooctye donors simplifies oocyte donation. AB - BACKGROUND: We report our experience on the efficacy of a new regimen of the GnRH antagonist, cetrorelix, and recombinant FSH, Gonal-F, for controlled ovarian stimulation in a donor oocyte programme. METHODS AND RESULTS: Six oocyte donors were commenced on Gonal-F (150 IU) and two on Gonal-F 225 IU daily on day 4 together with cetrorelix 0.25 mg daily on day 8 until the day of administration of hCG. Six premenopausal recipients were down-regulated with intranasal Nafarelin 400 micro g twice daily; two women with premature menopause did not require down-regulation for synchronization between donor and recipient cycles. The median (range) of oocytes retrieved and the median (range) fertilization rates were 7 (3-13) and 50% (0-71%) respectively. With the exception of a recipient who had failed fertilization, seven recipients had two embryos transferred. The median (range) number of days of ovarian stimulation, cetrorelix administration and number of Gonal-F ampoules administered for ovarian stimulation were 9 (7-12) days, 5 (3-8) and 18 (14-24) respectively. The clinical pregnancy rate per cycle was 50% (4/8) and one of the latter women miscarried at eight weeks gestation. Three women (37.3%) had full term deliveries. CONCLUSION: This preliminary study has shown that using a combination of cetrorelix and Gonal F resulted in a high pregnancy rate, reduced the duration of treatment for the donor and simplified oocyte donation. PMID- 12773456 TI - Concentrations of carotenoids, retinol and alpha-tocopherol in plasma and follicular fluid of women undergoing IVF. AB - BACKGROUND: Carotenoids are transferred into follicular fluid where they might serve as antioxidants and/or as precursors of retinoids which might modulate follicular or oocyte functions. METHODS AND RESULTS: In 77 women undergoing IVF differences between plasma and follicular fluid in the levels of carotenoids, retinol and alpha-tocopherol were evaluated especially with regard to fertilization success. Concentration of total carotenoids, retinol and alpha tocopherol determined by HPLC in follicular fluid and plasma were 0.06 +/- 0.02 versus 0.56 +/- 0.23 micro mol/l, 1.26 +/- 0.52 versus 1.66 +/- 0.37 micro mol/l and 4.89 +/- 2.13 versus 21.0 +/- 5.7 micro mol/l (mean +/- SD) respectively (P < 0.001 for all). Differences between plasma and follicular fluid were greater for beta-carotene and lycopene (<20% of plasma concentration) than for lutein and zeaxanthin (>40%). Intrafollicular retinol and retinol-binding protein (RBP) levels represented 58 +/- 23% and 60 +/- 19% of plasma level. Similar molar ratios of retinol/RBP were observed. While no differences in the mean values of all components investigated were observed in plasma and follicular fluid between women with and without reproductive success, the variability in the concentration was much greater in follicular fluid obtained from women without reproductive success. CONCLUSIONS: It remains to be elucidated, if this is indicative of a disturbed sieving effect of the blood-follicle barrier with possibly negative consequences for oocyte maturation. PMID- 12773457 TI - Oxidative DNA damage in human sperm influences time to pregnancy. AB - BACKGROUND: Oxidative stress and related DNA damage in human sperm may be important for fecundity and pregnancy outcome. METHODS: We studied the level of oxidative DNA damage in terms of 7-hydro-8-oxo-2'-deoxyguanosine (8-oxodG) in sperm DNA among 225 first-pregnancy planners. Over the six menstrual cycle follow up time, after cessation of contraception, 135 pregnancies were conceived. RESULTS: The likelihood of pregnancy occurring in a single menstrual cycle was inversely associated with the 8-oxodG level (P < 0.01). The odds ratio of pregnancy in each of the first three or all six follow-up menstrual cycles was 0.42 (0.23-0.78; 95% CI) and 0.61 (0.36-0.91) per unit increase in the log 8 oxodG/100 000 dG ratio after adjustment for potential confounders, (including sperm concentration) respectively. The intra-individual coefficient of variation of 8-oxodG in 2-6 monthly repeated sperm samples from 116 men was 19% for the 8 oxodG/dG ratio, whereas the inter-individual coefficient of variation was 49%. The 8-oxodG level was not significantly associated with smoking, consumption of alcohol or caffeine, exposure to welding fumes or the plasma levels of sex hormones. CONCLUSIONS: The data suggest that oxidative damage to sperm DNA influences fecundity and the level of damage is relatively constant within an individual and not influenced by smoking. PMID- 12773458 TI - Pregnancy by the tubal transfer of embryos developed after injection of round spermatids into oocyte cytoplasm of the cynomolgus monkey (Macaca fascicularis). AB - BACKGROUND: Round spermatids have been used as substitute gametes in basic reproductive research and in infertility clinics. In humans, however, the efficiency of fertilization and pregnancy is generally much lower after round spermatid injection (ROSI) than after injection with mature sperm. We examined the ability of round spermatids to support embryonic development using a non human primate as a model. We chose cynomolgus monkeys because, as in humans, their round spermatids have the oocyte-activating capacity of mature sperm. METHODS: We examined fertilization and subsequent development of embryos after ROSI and then transferred the embryos into the oviducts of female monkeys. RESULTS: Seventy-seven per cent of survived oocytes were activated and had formed pronuclei or the second polar body; 79% of the oocytes cultured developed to the 2-cell stage, and 23% developed to the blastocyst stage. Ultrasonography showed a normal-sized fetus in the uterus of a recipient, but the fetus spontaneously aborted at day 103. CONCLUSIONS: The round spermatids of cynomolgus monkeys can be used as substitute gametes to support embryonic development at least to mid gestation. This non-human primate is a suitable animal model for round spermatid conception in mammals, especially humans, and for biological and genetic characterization of events following ROSI. PMID- 12773459 TI - Testicular sperm extraction (TESE) and ICSI in patients with permanent azoospermia after chemotherapy. AB - BACKGROUND: Patients persistently azoospermic after chemotherapy have been considered traditionally as sterile unless sperm was frozen before therapy. Recent advances during the last decade combining testicular sperm extraction (TESE) and ICSI in patients with non-obstructive azoospermia allow these males to father their own genetic offspring. METHODS: A retrospective study was conducted of 12 patients with non-obstructive azoospermia after chemotherapy undergoing TESE between 1995 and 2002. Cancer type and anti-neoplastic treatments were recorded, together with maximum testicular volume, serum FSH levels and testicular histopathology. When TESE was successful, spermatozoa were cryopreserved for performing ICSI later. RESULTS: In five patients (41.6%) motile spermatozoa for cryopreservation and ICSI were retrieved. Four of them had received chemotherapy for testicular cancer, and one had been treated by chemotherapy/radiotherapy for Hodgkin's disease. Clinical and histological parameters were unable to predict with certainty TESE outcome in an individual patient. Eight ICSI cycles were performed on five couples and one pregnancy was obtained which resulted in the delivery of a healthy girl. CONCLUSION: Some patients with permanent azoospermia after chemotherapy can be successfully treated by TESE-ICSI. This procedure, however, may have potential genetic risks. Therefore, freezing semen before starting gonadotoxic therapy is the strategy of choice, and patients should be counselled accordingly. PMID- 12773460 TI - Different fertilization rates between immotile testicular spermatozoa and immotile ejaculated spermatozoa for ICSI in men with Kartagener's syndrome: case reports. AB - We report two cases of infertility treatment in couples where males suffered from Kartagener's syndrome (KS) and a total absence of motile sperm in the ejaculate. A total of three ICSI cycles was carried out. In all cycles, viable ejaculated or testicular spermatozoa were selected using the hypo-osmotic swelling (HOS) test. Case 1: In the first ICSI cycle total fertilization failure occurred after using ejaculated spermatozoa. In the following cycle testicular spermatozoa were used for ICSI, resulting in 75% fertilized oocytes and a pregnancy. Case 2: In the same ICSI cycle 50% of the oocytes were injected with ejaculated and 50% with testicular spermatozoa. The fertilization rates were 44 and 56% respectively and high quality embryos were achieved in both groups. One single embryo derived from testicular sperm was transferred with a resulting singleton pregnancy. In conclusion, testicular sperm for ICSI seem to have reliable fertilization capacity in men with KS, while ejaculated sperm, even if tested viable, seem more unpredictable. HOS test for selection of viable sperm for ICSI is recommended when ejaculated as well as testicular sperm are used for ICSI. PMID- 12773461 TI - Relationship between meiotic spindle location with regard to the polar body position and oocyte developmental potential after ICSI. AB - BACKGROUND: The recent development of a computer-assisted polarization microscopy system (Polscope) with which the meiotic spindle can be visualized in living oocytes on the basis of its birefringence permits analysis of the meiotic spindles of oocytes subjected to ICSI. Previous studies have shown that the meiotic spindle is not always aligned with the first polar body (PB) in metaphase II human oocytes prepared for ICSI. In the present study, the relationship between the degree of meiotic spindle deviation from the first PB location and ICSI outcome was analysed. METHODS: Oocytes were divided into four groups according to the angle of meiotic spindle deviation from the PB position. The angle of deviation was 0-5 degrees, 6-45 degrees, 46-90 degrees and >90 degrees for groups I to IV respectively. RESULTS: The rates of normal [2 pronuclei (PN)] and abnormal (1PN or >2PN) fertilization did not differ between groups I, II and III. However, the rate of normal fertilization was lower among oocytes in which the meiotic spindle deviation angle was >90 degrees; this led to an increased proportion of tripronucleated zygotes that failed to extrude the second PB. When embryos developed from normally fertilized oocytes were evaluated on day 3 after ICSI, no relationship was found between the angle of meiotic spindle deviation and embryo quality. The meiotic spindle was not detected in only 9% of oocytes, and these showed a higher incidence of fertilization and cleavage abnormalities than did oocytes in which the spindle was detected. When oocytes at metaphase I after cumulus oophorus and corona radiata removal were matured in vitro, the meiotic spindle was detected in 53.8% of those that reached metaphase II. In these in-vitro-matured oocytes the meiotic spindle was always aligned with the first PB, suggesting that misalignment seen in those oocytes matured in vivo resulted from PB displacement during manipulations for cumulus and corona removal. CONCLUSION: High degrees of misalignment between the meiotic spindle and the first PB predict an increased risk of fertilization abnormalities. However, when normal fertilization had occurred, the cleavage potential of embryos developing from such oocytes was not impaired. These findings facilitate the selection of oocytes for ICSI in situations when the creation of supernumerary embryos is to be avoided. PMID- 12773462 TI - Developmental competence of oocytes showing increased cytoplasmic viscosity. AB - BACKGROUND: The objective of the study was to investigate the developmental fate of oocytes with increased cytoplasmic viscosity as assessed by the persistence of the injection funnel after withdrawal of the ICSI pipette. METHODS: For this purpose, 1008 oocytes showing a characteristic injection funnel during ICSI were subdivided into two groups according to the oocyte's ability to restore its spherical shape within 2-3 min after ICSI. Fertilization and further development was evaluated in both groups. In addition, implantation and pregnancy rates were analysed. RESULTS: In the funnel positive cohort (group 1) significantly fewer oocytes degenerated after injection (P < 0.01) compared with oocytes without persistent funnel (group 2). However, at zygote stage, presence of a halo (P < 0.05) and a optimal pronuclear pattern 0 (P < 0.01) was increased in group 2. In addition, significantly fewer poor quality embryos were found in this group (P < 0.01). The number of good quality blastocysts but not blastocyst formation was increased in group 2 (P < 0.05). This resulted in an increased clinical pregnancy rate if embryos which derived exclusively from funnel negative oocytes were transferred (P < 0.05). CONCLUSIONS: Our data suggest that cytoplasm of higher viscosity delays development up to cleavage stage and impairs optimal development. Injection funnel persistence was found to be a negative prognostic marker of preimplantation development. PMID- 12773463 TI - The predictive value of using a combined Z-score and day 3 embryo morphology score in the assessment of embryo survival on day 5. AB - BACKGROUND: The purpose of this study was to evaluate the ability of using the Z score alone, or, in combination with the day 3 embryo morphology score, to predict embryo viability at day 5 from a large cohort of embryos derived from patients undergoing treatment with IVF/ICSI. METHODS AND RESULTS: In this retrospective study, a total of 1894 zygotes from 346 treatment cycles (295 couples) was analysed between January 2001 and May 2002. The Z-scoring system was useful in predicting day 5 embryo survival. The mean +/- SD day 5 embryo survival rates were 78.2 +/- 1.7, 49.0 +/- 2.5, 21.4 +/- 3.2 and 11.8 +/- 5.6% for Z-1, Z 2, Z-3 and Z-4 zygotes groups respectively. Embryos derived from Z-1 scores and grade I day 3 embryo scores showed the best day 5 embryo survival and a very high implantation potential. CONCLUSIONS: These data suggest that a combined evaluation of the Z-score and day 3 embryo morphology is highly predictive of embryo outcome after IVF/ICSI. The Z-score could be of great help in the selection of embryos for cultures extended to later stages. The Z-score alone, or preferably in combination with day 3 embryo morphology, is useful in the determination of the most suitable embryos and the number of embryos for transfer, thus achieving the optimal chance of conception while reducing the risk of high order multiple pregnancy. PMID- 12773464 TI - Sequential assessment of individually cultured human embryos as an indicator of subsequent good quality blastocyst development. AB - BACKGROUND: It is of fundamental importance for IVF clinics to determine the most viable embryos for transfer. The challenge for ART clinics is to transfer fewer embryos, thereby minimizing the risk of multiple-infant births, while still maintaining the greatest chance of pregnancy for their patients. In this study, an investigation was made to determine if developmental markers on the day of fertilization (day 1) can predict good subsequent blastocyst development. METHODS AND RESULTS: A total of 1550 individually cultured 2PN embryos from 191 patients undergoing IVF/ICSI treatment at the Yale University Center for Reproductive Medicine and Infertility from February to December 2001 was included. The results showed a significant positive relationship between early-cleaving 2-cell embryos and subsequent good quality > or =4-cell, > or =7-cell and blastocyst development (P < 0.05). PN symmetry (the relative size of the PN to each other), when checked at the time fertilization, is also a significant indictor of good quality > or =4 cell, > or =7-cell stage embryos and blastocysts. Combined, a developing embryo showing PN symmetry with early cleavage and subsequent good > or =4-cell and > or =7-cell cleavage, has a one in two chance of developing into a good-quality blastocyst. CONCLUSION: Early embryo assessment can be used as an indicator of subsequent good blastocyst development. PMID- 12773465 TI - Cryo-thawed embryos obtained from conception cycles have double the implantation and pregnancy potential of those from unsuccessful cycles. AB - BACKGROUND: The purpose of this study was to evaluate the influence of fresh IVF/ICSI cycle outcome on the prognosis of the related frozen embryo replacement (FER) cycle. METHODS: 459 FER cycles, involving 2049 cleavage stage embryos with no or up to 10% fragmentation, were performed for which the outcome of the fresh cycle was recorded. The cycles were divided into two groups; group A included cycles in which cryopreserved embryos were obtained from fresh cycles in which conception occurred. Group B were cycles in which cryopreserved embryos originated from unsuccessful fresh cycles. RESULTS: Groups A and B were comparable with respect to mean (+/- SD) age at cryopreservation (33 +/- 3.9 versus 33.2 +/- 4 years, P = not significant), mean number of oocytes retrieved and fertilized normally in the fresh cycle (11 +/- 5.2 versus 11.2 +/- 4.8, P = not significant) and mean age at the cryo-thawed transfer (34.5 +/- 4.2 versus 33.9 +/- 4 years, P = not significant). No significant difference was found between the two groups with regard to mean number of embryos cryopreserved (6.5 +/- 3.9 versus 6.2 +/- 3.6) and subsequently thawed (4.5 +/- 2.5 versus 4.5 +/- 1.8) per cycle and number of cryo-thawed embryos transferred per cycle (2.0 +/- 0.7 versus 2.1 +/- 0.8). However, the implantation rate per transferred embryo in group A was double that in group B (23 versus 11.2%, P < 0.0001). Moreover, the clinical pregnancy and ongoing pregnancy rates per cycle were significantly higher in group A compared with group B (34.8 and 27.3% versus 15.6 and 13.1%, P < 0.0001 and P = 0.0003 respectively). The difference in FER cycle outcome could not be explained by confounding variables. CONCLUSIONS: After thawing, cryopreserved embryos originating from conception IVF/ICSI cycles achieve double the implantation and pregnancy rates of those obtained from unsuccessful cycles. PMID- 12773466 TI - Normal birth after microsurgical enucleation of tripronuclear human zygotes: case report. AB - Microsurgical enucleation of a single pronucleus from each of three tripronuclear zygotes was performed and the embryos were transferred to a 38-year-old woman on day 3 after fertilization. A normal healthy baby boy was born at 38 weeks and 4 days gestation, demonstrating that with polyspermic fertilization, removal of the extra male pronucleus allows the zygote to develop normally. PMID- 12773467 TI - Histopathological extent of rectal invasion by rectovaginal endometriosis. AB - BACKGROUND: We aimed to evaluate the microscopic extent of endometriosis in surgical en-bloc specimens of vaginal skin, rectovaginal septum, cul-de-sac, and part of the rectosigmoid bowel. METHODS: From December, 1997 to October, 2001, 50 patients with the trias of intestinal pain, palpable disease in the rectovaginal septum, and laparoscopic diagnosis of endometriosis of the cul-de-sac and/or rectosigmoid colon underwent combined laparoscopic-vaginal en-bloc resection of the cul-de-sac with partial resection of the posterior vaginal wall and rectum with reanastomosis by minilaparotomy. All surgical specimens were histopathologically evaluated in a standardized fashion. RESULTS: The mean length of the bowel specimen was 7.48 cm. Endometriosis involved the serosa and muscularis propria in all patients, the submucosa in 17 patients (34%), and the mucosa in five patients (10%). After a mean follow-up of 32 months, 90% of patients reported a considerable improvement or were completely free of symptoms and the rate of recurrence was 4% (two patients). CONCLUSIONS: Partial bowel resection indicates the depth and multifocality of endometriosis affecting the recto-sigmoid colon. Such extensive surgery appears justified by the extent of the lesions and the long-term relief of symptoms achieved. PMID- 12773468 TI - Characteristic images of deeply infiltrating rectosigmoid endometriosis on transvaginal and transrectal ultrasonography. AB - BACKGROUND: To evaluate the usefulness of transvaginal and transrectal ultrasonography for diagnosis and management of deeply infiltrating rectosigmoid endometriosis. METHODS: A series of six patients (aged 32-39 years) with rectosigmoid endometriosis underwent transvaginal and transrectal ultrasonography. In three patients undergoing surgical resection of the intestine, the ultrasonographic findings were compared with macroscopic and microscopic findings. In one patient, sequential observations of the lesion using ultrasonography were conducted before and after medical treatment and following childbirth. RESULTS: In all cases, the lesion was detected as a hypoechoic irregular-shaped area surrounded by a hyperechoic rim located posterior to the uterus, with size ranging from 18 x 17 to 29 x 28 mm in diameter. The comparison of the ultrasonographical findings with histology revealed that the hypoechoic irregular-shaped area corresponded to a layer of hypertrophic muscularis propria of the lesion, while the hyperechoic rim represented the layer including the mucosa, submucosa and serosa. In one patient, the lesion decreased in size and lost its central hypoechoic area after childbirth in association with pain relief. CONCLUSIONS: Transvaginal and transrectal ultrasonography provides characteristic appearances for rectosigmoid endometriosis that correlate well with its histological findings. The procedures would be useful in the management of rectosigmoid endometriosis. PMID- 12773469 TI - Surrogacy: the experience of commissioning couples. AB - BACKGROUND: Findings are presented of a study of families with a child created through a surrogacy arrangement. This paper focuses on the commissioning couples' reports of their experiences. METHODS: A total of 42 couples with a 1-year-old child born through surrogacy were assessed using a standardized semi-structured interview. Data were obtained on motivations for surrogacy, details about the surrogate mother, experience of surrogacy during pregnancy and after birth and disclosure of the surrogacy to friends and family. RESULTS: Couples had considered surrogacy only after a long period of infertility or when it was the only option available. Couples retrospectively recalled their levels of anxiety throughout the pregnancy as low, and relationships between the couple and the surrogate mother were found to be generally good. This was the case regardless of whether or not the couple had known the surrogate mother prior to the arrangement. After the birth of the child, positive relations continued with the large majority of couples maintaining some level of contact with the surrogate mother. All couples had told family and friends about the surrogacy and were planning to tell the child. CONCLUSIONS: Commissioning couples generally perceived the surrogacy arrangement as a positive experience. PMID- 12773470 TI - Determining access to assisted reproductive technology: reactions of clinic directors to ethically complex case scenarios. AB - BACKGROUND: Our aim was to increase understanding of how patient selection is handled by assisted reproductive technology (ART) clinicians. METHODS: Ethically complex case scenarios were evaluated by the directors of USA ART clinics. Scenarios included using a son as sperm donor for his father, sex selection without associated disease, treatment of morally irresponsible couples, and a dispute over embryo disposition. Respondents reviewed eight scenarios and gave their opinions on whether to offer treatment. Reasons given for these decisions were placed into one of 13 categories. RESULTS: Survey response rate was 57%. Between 3 and 50% of respondents would treat in each case. Of reasons given, 'conditional' responses (requiring counselling, blood tests or agreement to other 'conditions') were common (31.4%). Non-maleficence (risk) accounted for 29.4% of responses, philosophy of medicine 18.9%, respect for patient autonomy 5.9% and legal concerns 4.6%. Discrimination and threats were each significant in one case. Reasons evoking absolutist beliefs, personal discomfort, commitment to justice, religion and ethical relativism were rare. CONCLUSIONS: Clinicians felt conflict between a desire to respect patient autonomy and their discomfort over the risk associated with the procedure. They raised concerns about misuse of medical technology. Attempts to resolve complex issues through negotiation and compromise were common. PMID- 12773471 TI - Donation of embryos for stem cell research--how many couples consent? AB - BACKGROUND: The huge potential of human embryonic stem cells has been a subject of wide discussion as regards the ethical and legal justification of using human embryos for establishing such cell lines. The opinions of infertile couples and their willingness to donate their supernumerary embryos for stem cell research have not been investigated earlier. METHODS: We conducted an analysis of the answers of couples who were asked to give informed consent as regards donating their embryos for stem cell research in our IVF unit in 2001-2002. RESULTS: Ninety-two percent of the couples gave informed consent as regards establishing and characterizing embryonic stem cell lines from the embryos which could not be used in their infertility treatment. Discussion in the Swedish media during May to December, 2001 regarding the importance and ethical justification of stem cell research made informing the couples easier. CONCLUSION: A high proportion, 92%, of couples who underwent infertility treatment in Sweden preferred donating their supernumerary embryos for stem cell research rather than letting them be discarded. PMID- 12773472 TI - Effects of LH on oocyte yield and developmental competence. PMID- 12773474 TI - A health-economic decision-analytic model comparing double with single embryo transfer in IVF/ICSI: a sensitivity analysis. PMID- 12773475 TI - Detailed glycan analysis of serum glycoproteins of patients with congenital disorders of glycosylation indicates the specific defective glycan processing step and provides an insight into pathogenesis. AB - The fundamental importance of correct protein glycosylation is abundantly clear in a group of diseases known as congenital disorders of glycosylation (CDGs). In these diseases, many biological functions are compromised, giving rise to a wide range of severe clinical conditions. By performing detailed analyses of the total serum glycoproteins as well as isolated transferrin and IgG, we have directly correlated aberrant glycosylation with a faulty glycosylation processing step. In one patient the complete absence of complex type sugars was consistent with ablation of GlcNAcTase II activity. In another CDG type II patient, the identification of specific hybrid sugars suggested that the defective processing step was cell type-specific and involved the mannosidase III pathway. In each case, complementary serum proteome analyses revealed significant changes in some 31 glycoproteins, including components of the complement system. This biochemical approach to charting diseases that involve alterations in glycan processing provides a rapid indicator of the nature, severity, and cell type specificity of the suboptimal glycan processing steps; allows links to genetic mutations; indicates the expression levels of proteins; and gives insight into the pathways affected in the disease process. PMID- 12773476 TI - Glycosylation efficiency of Asn-Xaa-Thr sequons is independent of distance from the C-terminus in membrane dipeptidase. AB - In vitro transcription/translation studies with model proteins have shown that glycosylation of Asn-Xaa-Thr sequons is reduced when the sequon is within 60 residues of the C-terminus of the protein. We have previously shown that in living cells N-glycosylation of the prion protein (PrP) is also abolished when its Asn-Ile-Thr and Asn-Phe-Thr sequons are less than 60 residues from the C terminus (Walmsley and Hooper [2003] Biochemical Journal, 370, 351-355). To investigate whether sequon distance to the C-terminus is a general determinant of N-glycosylation in living cells, Asn-Ile/Phe-Thr sequons were introduced into another glycosylphosphatidylinositol (GPI) anchored protein, membrane dipeptidase (MDP), at similar distances from the C-terminus as those in PrP. When expressed in the human neuroblastoma SH-SY5Y cell line, the introduced sequons were fully N glycosylated even when they were less than 60 residues from the C-terminus in both GPI-anchored and secreted forms of MDP. These data demonstrate that the utilization of sequons in some proteins is independent of their distance from the C-terminus. PMID- 12773477 TI - Modulation of prion protein structural integrity by geldanamycin. AB - The cellular prion protein PrPc is of crucial importance for the development of neurodegenerative diseases called transmissible spongiform encephalopathies. We investigated if the function of members of the HSP90 family is required for the integrity of the normal, nonpathogenic prion protein called PrPc. Eukaryotic cells were treated with the structurally unrelated HSP90-inhibitors geldanamycin (GA) or radicicol (RC). In either case the cellular prion protein was induced and exhibited faster migrating bands on western blot analysis, whereas geldampicin (GE), an analog of GA known not to bind to HSP90, had no effect. Ongoing protein and messenger RNA synthesis during treatment were found to be necessary for the appearance of these bands. Cotreatment with tunicamycin abrogated any effect of HSP90 inhibitors on the cellular prion protein. Finally, enzymatic deglycosylation with peptide:N-glycosidase F of the normal prion protein as well as the variant induced by benzoquinone ansamycins resulted in very similar band patterns. These experiments indicate that either altered glycosylation, or a change in conformation, or both are involved in the induction of faster migrating bands by HSP90 inhibitors. Thus the inhibition of the function of members of the HSP90 family of molecular chaperones results in profound changes in the physicochemical properties of PrPc. PMID- 12773478 TI - Improved and simple micro assay for sulfated glycosaminoglycans quantification in biological extracts and its use in skin and muscle tissue studies. AB - This article describes a simple and selective procedure used for direct measurement of sulfated glycosaminoglycans (GAGs) in biological samples and its application to the determination of GAGs during tissue regeneration and myogenic differentiation. We describe a modified procedure of previous GAG assays that has improved specificity, reproducibility, and sensitivity. The assay is based on the ability of sulfated GAGs to bind the cationic dye 1,9-dimethylmethylene blue. We describe conditions that allow isolation of the GAG-dye complex. This complex was dissociated; the optical density measurement of the dissociated dye permitted quantification of GAGs in biological samples. Applied to the study of myogenic cell differentiation in vitro, muscle repair, and skin ulceration, this method revealed significant modifications in the patterns of expression of different sulfated GAGs in these tissues. In particular, application of the method after nitrous acid treatment revealed that heparan sulfate and chondroitin sulfate ratio changed during muscle regeneration process. PMID- 12773479 TI - Binding of the CC-chemokine RANTES to syndecan-1 and syndecan-4 expressed on HeLa cells. AB - It is believed that proteoglycans influence biological properties of chemokines. We show that the CC chemokine RANTES binds not only to high-affinity binding sites on CCR5-positive HeLa cells but also to low-affinity binding sites on HeLa cells expressing or lacking RANTES G protein-coupled receptors. Coimmunoprecipitation studies demonstrate that RANTES forms complexes with glycanated syndecan (SD)-1 and -4, in addition to CCR5 on the CCR5-positive HeLa cells. Moreover, confocal microscopy analysis shows the colocalization of RANTES with SD-1 and -4. Glycosaminoglycans removal from the cells by glycosaminidases treatment prevented RANTES binding to SD-1 and -4 and decreased RANTES binding to CCR5 on the CCR5-positive HeLa cells. Removal of glycosaminoglycans by glycosaminidases treatment of the complexes, RANTES/SD-1/SD-4/+/-CCR5, immobilized on beads, reversed SD-1 and -4 bindings. Therefore, RANTES bindings to SD-1 and -4 depend on glycosaminoglycans and facilitate RANTES interaction with CCR5. Extracting plasma membrane cholesterol abolished the coimmunoprecipitation of SD-1 with RANTES, suggesting that rafts are involved in RANTES association to SD-1. Confocal microscopy analysis as well as coimmunoprecipitation experiments show a RANTES-independent heteromeric complex on the CCR5-positive HeLa cells, SD-1, SD-4, and CCR5. This complex is likely a functional unit in which proteoglycans may modulate RANTES binding to CCR5. PMID- 12773480 TI - The dsRNA binding protein family: critical roles, diverse cellular functions. AB - The dsRNA binding proteins (DRBPs) comprise a growing family of eukaryotic, prokaryotic, and viral-encoded products that share a common evolutionarily conserved motif specifically facilitating interaction with dsRNA. Proteins harboring dsRNA binding domains (DRBDs) have been reported to interact with as little as 11 bp of dsRNA, an event that is independent of nucleotide sequence arrangement. More than 20 DRBPs have been identified and reportedly function in a diverse range of critically important roles in the cell. Examples include the dsRNA-dependent protein kinase PKR that functions in dsRNA signaling and host defense against virus infection and DICER, which is implicated in RNA interference (RNAi) -mediated gene silencing. Other DRBPs such as Staufen, adenosine deaminase acting on RNA (ADAR), and spermatid perinuclear RNA binding protein (SPNR) are known to play essential roles in development, translation, RNA editing, and stability. In many cases, homozygous and even heterozygous disruption of DRBPs in animal models results in embryonic lethality. These results implicate the recognition of dsRNA as an evolutionarily conserved mechanism important in the regulation of gene expression and in host defense and underscore the diversity of essential biological tasks performed by dsRNA-related processes in the cell. PMID- 12773481 TI - CD105 is important for angiogenesis: evidence and potential applications. AB - Angiogenesis is the propelling force for tumor growth and metastasis, and antiangiogenic therapy represents one of the most promising modalities for cancer treatment. CD105 (endoglin) is a proliferation-associated and hypoxia-inducible protein abundantly expressed in angiogenic endothelial cells (EC). It is a receptor for transforming growth factor (TGF) -beta1 and -beta3 and modulates TGF beta signaling by interacting with TGF-beta receptors I and/or II. Immunohistochemistry studies have revealed that CD105 is strongly expressed in blood vessels of tumor tissues. Intratumoral microvessel density (MVD) determined using antibodies to CD105 has been found to be an independent prognostic indicator, wherein increased MVD correlates with shorter survival. CD105 is able to be shed into the circulation, with elevated levels detected in patients with various types of cancer and positively correlated with tumor metastasis. Tangible evidence of its proangiogenic role comes from knockout studies in which CD105 null mice die in utero as a result of impaired angiogenesis in the yolk sac and heart defects. The potential usefulness of CD105 for tumor imaging has been evaluated in tumor-bearing mice and dogs that have shown the rapid accumulation of radiolabeled anti-CD105 monoclonal antibody in the tumors with a high tumor-to background ratio. The anti-CD105 antibody conjugated with immunotoxins and immunoradioisotopes efficiently suppressed/abrogated tumor growth in murine models bearing breast and colon carcinoma without any significant systemic side effects. Immunoscintigraphy in patients with renal cell carcinomas has shown specific localization of 99Tcm-labeled CD105 mab in tumor endothelial cells. Thus, CD105 is a promising vascular target that can be used for tumor imaging, prognosis, and bears therapeutic potential in patients with solid tumors and other angiogenic diseases. PMID- 12773482 TI - Recovery of hepatocellular ATP and "pericentral apoptosis" after hemorrhage and resuscitation. AB - Progressive liver dysfunction contributes significantly to the development of multiple organ failure after trauma/hemorrhage. This study tested the relative impact of necrotic and apoptotic cell death in a graded model of hemorrhagic shock (mean arterial blood pressure=35+/-5 mmHg for 1, 2, or 3 h, followed by 2 h, 1 h, or no resuscitation, respectively) in rats. Prolonged periods of hemorrhagic hypotension (3 h) were paralleled by a profound decrease of hepatic ATP levels and occurrence of pericentral necrosis. Resuscitation after shorter periods of hemorrhagic hypotension resulted in restoration of tissue ATP whereas hepatocellular function as assessed by indocyanine green clearance remained depressed (49.9+/-1.6 mL/(min x kg) at baseline, 28.8+/-1.2 mL/(min x kg) after 2 h of resuscitation; P<0.05). Under these conditions, induction of caspase activity and DNA fragmentation were observed in pericentral hepatocytes that could be prevented by the radical scavenger tempol. Pretreatment with z-Val-Ala Asp(O-methyl)-flouromethylketone prevented de novo expression of caspase generated cytokeratin 18, DNA fragmentation, and depression of hepatocellular indocyanine green clearance. These data suggest that prolonged low flow/hypoxia induces ATP depletion and pericentral necrosis and restoration of oxygen supply and ATP levels after shorter periods of low flow ischemia propagate programmed cell death or "pericentral apoptosis." PMID- 12773483 TI - Regulation by complement C3a and C5a anaphylatoxins of cytokine production in human umbilical vein endothelial cells. AB - C3a and C5a anaphylatoxins are cytokine-like polypeptides generated during complement (C) system activation and released at the inflammatory site. They exert several biological activities through binding to the G-protein-coupled receptors C3aR and C5aR, respectively. Cloning and Northern blot experiments have indicated that both receptors are expressed by myeloid as well as nonmyeloid cells (e.g., endothelial and epithelial cells). To better understand the roles of C anaphylatoxins during inflammation, we investigated their effects on the expression of cytokine and chemokine genes by cultured human umbilical cord endothelial cells (HUVEC). HUVEC constitutively expressed both anaphylatoxin receptors, and addition of physiological concentrations of C3a or C5a (nM range) caused a strong up-regulation of IL-8, IL-1beta, and RANTES mRNA in a time- and dose-dependent manner. Conversely, a decrease in IL-6 mRNA was observed, but only with C5a stimulation. These variations in mRNA levels were inhibited by pretreatment with anti-C5aR and anti-C3aR antibodies as well as pertussis toxin, indicating that G-proteins are involved in anaphylatoxin-activated signal transduction pathways. Finally, we showed that C3a and C5a both strongly activate downstream MAP kinase signaling pathways (p44 and p42 Erk kinases). PMID- 12773484 TI - Heparanase mediates cell adhesion independent of its enzymatic activity. AB - Heparanase is an endo-beta-D-glucuronidase that cleaves heparan sulfate and is implicated in diverse physiological and pathological processes. In this study we report on a novel direct involvement of heparanase in cell adhesion. We demonstrate that expression of heparanase in nonadherent lymphoma cells induces early stages of cell adhesion, provided that the enzyme is expressed on the cell surface. Heparanase-mediated cell adhesion to extracellular matrix (ECM) results in integrin-dependent cell spreading, tyrosine phosphorylation of paxillin, and reorganization of the actin cytoskeleton. The surface-bound enzyme also augments cell invasion through a reconstituted basement membrane. Cell adhesion was augmented by cell surface heparanase regardless of whether the cells were transfected with active or point mutated inactive enzyme, indicating that heparanase functions as an adhesion molecule independent of its endoglycosidase activity. The combined feature of heparanase as an ECM-degrading enzyme and a cell adhesion molecule emphasizes its significance in processes involving cell adhesion, migration, and invasion, including embryonic development, neovascularization, and cancer metastasis. PMID- 12773485 TI - T helper (Th) 2 predominance in atopic diseases is due to preferential apoptosis of circulating memory/effector Th1 cells. AB - T cells constitute a large population of cellular infiltrate in atopic/allergic inflammation and a dysregulated, Th2-biased peripheral immune response appears to be an important pathogenetic factor. In atopic dermatitis, circulating cutaneous lymphocyte-associated antigen-bearing (CLA+) CD45RO+ T cells with skin-specific homing property represent an activated memory/effector T cell subset. They express high levels of Fas and Fas ligand and undergo activation-induced apoptosis. The freshly purified CLA+ CD45RO+ T cells of atopic individuals display distinct features of in vivo-triggered apoptosis such as pro-caspase degradation and active caspase-8 formation. In particular, the Th1 compartment of activated memory/effector T cells selectively undergoes activation-induced cell death, skewing the immune response toward surviving Th2 cells in atopic dermatitis patients. The apoptosis of circulating memory/effector T cells was confined to atopic individuals whereas non-atopic patients such as psoriasis, intrinsic-type asthma, contact dermatitis, intrinsic type of atopic dermatitis, bee venom allergic patients, and healthy controls showed no evidence for enhanced T cell apoptosis in vivo. These results define a novel mechanism for peripheral Th2 response in atopic diseases. PMID- 12773486 TI - Nitric oxide prevents 6-hydroxydopamine-induced apoptosis in PC12 cells through cGMP-dependent PI3 kinase/Akt activation. AB - Nitric oxide (NO) functions not only as an important signaling molecule in the brain by producing cGMP, but also regulates neuronal cell apoptosis. The mechanism by which NO regulates apoptosis is unclear. In this study, we demonstrated that NO, produced either from the NO donor S-nitroso-N-acetyl-d,l penicillamine (SNAP) or by transfection of neuronal NO synthase, suppressed 6 hydroxydopamine (6-OHDA)-induced apoptosis in PC12 cells by inhibiting mitochondrial cytochrome c release, caspase-3 and -9 activation, and DNA fragmentation. This protection was significantly reversed by the soluble guanylyl cyclase inhibitor 1H-(1,2,4)-oxadiazole[4,3-a]quinoxalon-1-one, indicating that cGMP is a key mediator in NO-mediated anti-apoptosis. Moreover, the membrane permeable cGMP analog 8-Br-cGMP inhibited 6-OHDA-induced apoptosis. These anti apoptotic effects of SNAP and 8-Br-cGMP were suppressed by cGMP-dependent protein kinase G (PKG) inhibitor KT5823, indicating that PKG is a downstream signal mediator in the suppression of apoptosis by NO and cGMP. Both SNAP and 8-Br-cGMP induced endogenous Akt activation and Bad phosphorylation, resulting in the inhibition of Bad translocation to mitochondria; these effects were inhibited by KT5823 and the phosphatidylinositol 3-kinase (PI3K) inhibitors LY294002 and Wortmannin. Our data suggest that the NO/cGMP pathway suppresses 6-OHDA-induced PC12 cell apoptosis by suppressing the mitochondrial apoptosis signal via PKG/PI3K/Akt-dependent Bad phosphorylation. PMID- 12773487 TI - TNF-alpha increases ubiquitin-conjugating activity in skeletal muscle by up regulating UbcH2/E220k. AB - In some inflammatory diseases, TNF-alpha is thought to stimulate muscle catabolism via an NF-kappaB-dependent process that increases ubiquitin conjugation to muscle proteins. The transcriptional mechanism of this response has not been determined. Here we studied the potential role of UbcH2, a ubiquitin carrier protein and homologue of murine E220k. We find that UbcH2 is constitutively expressed by human skeletal and cardiac muscles, murine limb muscle, and cultured myotubes. TNF-alpha stimulates UbcH2 expression in mouse limb muscles in vivo and in cultured myotubes. The UbcH2 promoter region contains a functional NF-kappaB binding site; NF-kappaB binding to this sequence is increased by TNF-alpha stimulation. A dominant negative inhibitor of NF-kappaB activation blocks both UbcH2 up-regulation and the increase in ubiquitin conjugating activity stimulated by TNF-alpha. In extracts from TNF-alpha-treated myotubes, ubiquitin-conjugating activity is limited by UbcH2 availability; activity is inhibited by an antiserum to UbcH2 or a dominant negative mutant of UbcH2 and is enhanced by wild-type UbcH2. Thus, UbcH2 up-regulation is a novel response to TNF-alpha/NF-kappaB signaling in skeletal muscle that appears to be essential for the increased ubiquitin conjugation induced by this cytokine. PMID- 12773488 TI - Folding of HIV-1 envelope glycoprotein involves extensive isomerization of disulfide bonds and conformation-dependent leader peptide cleavage. AB - Human immunodeficiency virus binds and enters cells via the Envelope glycoprotein gp160 at its surface. In infected cells, gp160 is found not only on the plasma membrane but also in the endoplasmic reticulum (ER). Our aim was to establish rate-determining steps in the maturation process of gp160, using a radioactive pulse-chase approach. We found that gp160 has an intricate folding process: disulfide bonds start to form during synthesis but undergo extensive isomerization until the correct native conformation is reached. Removal of the leader peptide critically depends on formation of at least some disulfide bonds in subunit gp120 during folding. Envelope folds extremely slowly and therefore resides in the ER longer than other proteins, but the yield of properly folded molecules is high and degradation is undetectable. The large quantity of gp160 in the ER hence is a result of its slow transit through this compartment. We show here that newly synthesized HIV-1 Envelope glycoprotein apparently follows a slow but high-yield folding path in which co- and post-translational formation of disulfide bonds in gp120, disulfide isomerization and conformation dependent removal of the leader sequence are determining and intertwined events. PMID- 12773489 TI - Interaction of low molecular weight group IIA phospholipase A2 with apoptotic human T cells: role of heparan sulfate proteoglycans. AB - Human group IIA phospholipase A2 (hIIA PLA2) is a 14 kDa secreted enzyme associated with inflammatory diseases. A newly discovered property of hIIA PLA2 is the binding affinity for the heparan sulfate proteoglycan (HSPG) glypican-1. In this study, the binding of hIIA PLA2 to apoptotic human T cells was investigated. Little or no exogenous hIIA PLA2 bound to CD3-activated T cells but significant binding was measured on activated T cells induced to undergo apoptosis by anti-CD95. Binding to early apoptotic T cells was greater than to late apoptotic cells. The addition of heparin and the hydrolysis of HSPG by heparinase III only partially inhibited hIIA PLA2 binding to apoptotic cells, suggesting an interaction with both HSPG and other binding protein(s). Two low molecular weight HSPG were coimmunoprecipitated with hIIA PLA2 from apoptotic T cells, but not from living cells. Treatment of CD95-stimulated T cells with hIIA PLA2 resulted in the release of arachidonic acid but not oleic acid from cells and this release was blocked by heparin and heparinase III. Altogether, these results suggest a role for hIIA PLA2 in the release of arachidonic acid from apoptotic cells through interactions with HSPG and its potential implication in the progression of inflammatory diseases. PMID- 12773490 TI - Methylation status of uPA promoter as a molecular mechanism regulating prostate cancer invasion and growth in vitro and in vivo. AB - Urokinase plasminogen activator (uPA) promotes tumor invasion and metastasis in several malignancies including prostate cancer, one of the most commonly detected male cancers that result in a high incidence of mortality. In the present study we have examined the differential regulation of uPA gene expression in different stages of prostate cancer by DNA methylation. We determined levels of uPA expression in normal prostate epithelial cells (PrEC) and in hormone-responsive (LNCaP) and -insensitive (PC-3) prostate cancer cell lines. We found that uPA is expressed only in the highly invasive PC-3 cells where the uPA promoter is unmethylated. The lack of uPA expression in PrEC and LNCaP cells, where uPA promoter is highly methylated, is due to suppression of uPA gene transcription by DNA methylation. Treatment of LNCaP cells with 5'-azacytidine, a potent demethylating agent, resulted in induction of uPA mRNA expression, uPA activity, and higher invasive capacity in vitro. Additionally, a marked increase in tumor volume was observed after inoculation of these cells into the flank of male BALB/c (nu/nu) mice. Collectively these studies have demonstrated that DNA methylation is the underlying molecular mechanism responsible for uPA gene silencing in normal and early stages of prostate cancer, which has a direct effect on tumor cell invasion and growth in vitro and in vivo. PMID- 12773491 TI - Histamine enhances TGF-beta1-mediated suppression of Th2 responses. AB - Susceptibility of T cells to TGF-beta1 produced by regulatory T cells has an important impact on the induction and maintenance of peripheral tolerance and therefore on the development of autoimmunity, cancer, and allergy. Histamine not only mediates the deleterious effects of allergic reactions, it can also modulate the Th1/Th2 cell balance. We demonstrate that histamine dose-dependently enhanced TGF-beta1-mediated suppression and TGF-beta1 responsiveness of CD4+ T cells. This effect was mediated by the histamine 2 receptor (H2R), as demonstrated by receptor-specific agonists and antagonists. Furthermore, the histamine effect on TGF-beta1 responsiveness was cAMP/PKA dependent. This pathway is activated by the H2R, which is preferentially expressed on Th2 cells. Thus a higher additive effect of histamine on TGF-beta1 responsiveness was found in Th2 cells compared with Th1 cells. In fact, findings are confirmed by analysis of cytokine regulation, since activation of the H2R/cAMP pathway promoted TGF-beta1-mediated IL-4 inhibition but was ineffective in suppressing IFN-gamma. These results demonstrate that histamine supports TGF-beta1 susceptibility of T cells. Moreover, Th2 cells are more affected by histamine-enhanced TGF-beta1 suppression, which is particularly important for the regulation of allergen specific T cells in allergic immune responses. PMID- 12773492 TI - Role of development in reorganization of the SI forelimb-stump representation in fetally, neonatally, and adult amputated rats. AB - Rats that sustain forelimb removal on postnatal day (P) 0 exhibit numerous multi unit recording sites in the forelimb-stump representation of primary somatosensory cortex (SI) that also respond to hindlimb stimulation when cortical GABAA+B receptors are blocked. Most of these hindlimb inputs originate in the medial SI hindlimb representation. Although many forelimb-stump sites in these animals respond to hindlimb stimulation, very few respond to stimulation of the face (vibrissae or lower jaw), which is represented in SI just lateral to the forelimb. The lateral to medial development of SI may influence the capacity of hindlimb (but not face) inputs to "invade" the forelimb-stump region in neonatal amputees. The SI forelimb-stump was mapped in adult (>60 days) rats that had sustained amputation on embryonic day (E) 16, on P0, or during adulthood. GABA receptors were blocked and subsequent mapping revealed increases in nonstump inputs in E16 and P0 amputees: fetal amputees exhibited forelimb-stump sites responsive to face (34%), hindlimb (10%), and both (22%); neonatal amputees exhibited 10% face, 39% hindlimb, and 5% both; adult amputees exhibited 10% face, 5% hindlimb, and 0% both, with approximately 80% stump-only sites. These results indicate age-dependent differences in receptive-field reorganization of the forelimb-stump representation, which may reflect the spatiotemporal development of SI. Results from cobalt chloride inactivation of the SI vibrissae region and electrolesioning of the dysgranular cortex suggest that normally suppressed vibrissae inputs to the SI forelimb-stump area originate in the SI vibrissae region and synapse in the dysgranular cortex. PMID- 12773493 TI - Neural changes in cat auditory cortex after a transient pure-tone trauma. AB - Here we present the changes in cortical activity occurring within a few hours after a 1-h exposure to a 120-dB SPL pure tone (5 or 6 kHz). The changes in primary auditory cortex of 16 ketamine-anesthetized cats were assessed by recording, with two 8-microelectrode arrays, from the same multiunit clusters before and after the trauma. The exposure resulted in a peripheral threshold increase that stabilized after a few hours to on average 40 dB in the frequency range of 6-32 kHz, as measured by the auditory brain stem response. The trauma induced a shift in characteristic frequency toward lower frequencies, an emergence of new responses, a broadening of the tuning curve, and an increase in the maximum of driven discharges. In addition, the onset response after the trauma was of shorter duration than before the trauma. The results suggest the involvement of both a decrease and an increase in inhibition. They are discussed in terms of changes in central inhibition and its implications for tonotopic map plasticity. PMID- 12773494 TI - Sustained muscle contractions maintained by autonomous neuronal activity within the human spinal cord. AB - It is well known that muscle contraction can be easily evoked in the human soleus muscle by applying single-pulse electrical stimulation to the tibial nerve at the popliteal fossa. We herein reveal the unexpected phenomenon of muscle contractions that can be observed when train stimulation is used instead. We found, in 11 human subjects, that transient electrical train stimulation (1-ms pulses, 50 Hz, 2 s) was able to induce sustained muscle contractions in the soleus muscle that outlasted the stimulation period for greater than 1 min. Subjects were unaware of their own muscle activity, suggesting that this is an involuntary muscle contraction. In fact, the excitability of the primary motor cortex (M1) with the sustained muscle contractions evaluated by transcranial magnetic stimulation was lower than the excitability with voluntary muscle contractions even when both muscle contraction levels were matched. This finding indicates that M1 was less involved in maintaining the muscle contractions. Furthermore, the muscle contractions did not come from spontaneous activity of muscle fibers or from reverberating activity within closed neuronal circuits involving motoneurons. These conclusions were made based on the respective evidence: 1) the electromyographic activity was inhibited by stimulation of the common peroneal nerve that has inhibitory connections to the soleus motoneuron pool and 2) it was not abolished after stopping the reverberation (if any) for approximately 100 ms by inducing the silent period that followed an H-reflex. These findings indicate that the sustained muscle contractions induced in this study are most likely to be maintained by autonomous activity of motoneurons and/or interneurons within the human spinal cord. PMID- 12773495 TI - Role of nucleus tractus solitarius 5-HT3 receptors in the defense reaction induced inhibition of the aortic baroreflex in rats. AB - Different stressful conditions elicit a typical behavior called the defense reaction. Our aim was to determine whether 5-HT3 receptors in the nucleus tractus solitarius (NTS) are involved in 1) the inhibition of the baroreflex bradycardia and 2) the rise in blood pressure, which are known to occur during the defense reaction. In urethane-anesthetized rats, the defense reaction was elicited by electrical stimulation of the dorsomedial nucleus of the hypothalamus (DMH) or the dorsal part of the periaqueductal gray (dPAG). Direct electrical stimulation of the aortic depressor nerve was used to trigger the typical baroreflex responses. Aortic stimulation at high (100-150 microA) and low (50-90 microA) intensity produced a decrease in heart rate of -39 to -44% (relative to baseline, Group 1 responses, n = 113) and -19 to -24% (Group 2 responses, n = 43), respectively. In spontaneously breathing rats, Group 1 and Group 2 bradycardiac responses were inhibited during DMH (-75 +/- 4% and -96 +/- 4%, n = 38 and n = 11, respectively), as well as dPAG (-81 +/- 3% and -95 +/- 4%, n = 36 and n = 10, respectively) stimulation. The aortic baroreflex bradycardia was hardly affected by DMH or dPAG stimulation when bicuculline (5 pmol), a specific GABAA receptor antagonist, had previously been microinjected into the NTS. Likewise, NTS microinjections of granisetron, a specific 5-HT3 receptor antagonist, prevented, in a dose-dependent manner, the baroreflex bradycardia inhibition. In addition, intra-NTS granisetron did not affect the rise in blood pressure induced by either site stimulation. These data show that 5-HT3 receptors in the NTS are involved in the GABAergic inhibition of the aortic baroreflex bradycardia, but not in the rise in blood pressure, occurring during the defense reaction elicited by DMH or dPAG stimulation. PMID- 12773496 TI - Human cortical responses to water in the mouth, and the effects of thirst. AB - In an event-related functional magnetic resonance imaging (fMRI) study in humans it was shown, first, that water produces activations in cortical taste areas (in particular the frontal operculum/anterior insula which is the primate primary taste cortex, and the caudal orbitofrontal/secondary taste cortex) comparable to those produced by the prototypical tastants salt and glucose. Second, the activations in the frontal operculum/anterior insula produced by water when thirsty were still as large after the subjects had consumed water to satiety. Third, in contrast, the responses to water in the caudal orbitofrontal cortex were modulated by the physiological state of the body, in that responses to the oral delivery of water in this region were not found after the subjects had drunk water to satiety. Fourth, further evidence that the reward value or pleasantness of water is represented in the orbitofrontal cortex was that a positive correlation with the subjective ratings of the pleasantness of the water was found with activations in the caudal and anterior orbitofrontal cortex, and also in the anterior cingulate cortex. Fifth, it was found that a region of the middle part of the insula was also activated by water in the mouth, and further, that this activation only occurred when thirsty. Sixth, analyses comparing pre- and postsatiety periods (i.e., when thirsty and when not thirsty) independently of stimulus delivery revealed higher activity levels in the rostral anterior cingulate cortex. The activity of the rostral anterior cingulate cortex thus appears to reflect the thirst level or motivational state of the subjects. PMID- 12773497 TI - Vestibular contributions to gaze stability during transient forward and backward motion. AB - The accuracy with which the vestibular system anticipates and compensates for the visual consequences of translation during forward and backward movements was investigated with transient motion profiles in rhesus monkeys trained to fixate targets on an isovergence screen. Early during motion when visuomotor reflexes remain relatively ineffective and vestibular-driven mechanisms have an important role for controlling the movement of the eyes, a large asymmetry was observed for forward and backward heading directions. During forward motion, ocular velocity gains increased steeply and reached near unity gains as early as 40-50 ms after motion onset. In addition, instantaneous directional errors also remained <10 degrees for forward headings. In contrast, backward motion was characterized by smaller vestibular gains and larger directional errors during the first 70 ms of the movement. To evaluate the accumulated retinal slip and vergence errors during the early epochs of motion when vestibular-driven mechanisms dominate gaze stability, the movement of a virtual fixation point defined by the intersection of the two gaze lines was quantitatively compared with the respective movement of the extinguished target in head coordinates. Both conjugate retinal slip and vergence errors were <0.2 degrees during the first 70 ms of the movement, with forward motion conjugate errors typically being smaller as compared with backward motion directions. Thus vestibularly driven gaze stabilization mechanisms can effectively minimize conjugate retinal slip errors as well as keep binocular disparity errors low during the open loop interval of head movement. PMID- 12773498 TI - Experimental autoimmune autonomic neuropathy. AB - Antibodies specific for the neuronal ganglionic nicotinic acetylcholine receptor (nAChR) are found in high titer in serum of patients with subacute autonomic failure. This clinical disorder is known as autoimmune autonomic neuropathy (AAN). Rabbits immunized with a neuronal nAChR alpha3 subunit fusion protein produce ganglionic nAChR antibodies and develop autonomic failure (experimental AAN, or EAAN). We used quantitative measures of autonomic function to demonstrate that this animal model of neuronal nAChR autoimmunity recapitulates the cardinal autonomic features of AAN in humans. The severity of dysautonomia in the rabbit ranges from isolated cardiovagal impairment to severe panautonomic failure with fixed mydriasis, gastroparesis, dry eyes, impaired heart rate variability, hypotension, and low plasma catecholamines. The severity of autonomic failure correlates with serum antibody levels. Immunohistochemical staining of superior cervical ganglia and myenteric plexus neurons demonstrates intact presynaptic nerve terminals and intact postsynaptic neurons containing cytoplasmic nAChR, but lacking surface nAChR. These findings define the autonomic physiology and histopathology of this novel animal model and support the concept that AAN in humans is a disorder of ganglionic cholinergic synaptic transmission caused by ganglionic nAChR antibodies. PMID- 12773499 TI - Graded limb targeting in an insect is caused by the shift of a single movement pattern. AB - Grooming responses are movements of a multi-jointed limb that are targeted toward a stimulus site on the body. To be successful, they require a continuous transformation of stimulus location into a corresponding motor pattern or selection and blending of a subset of cardinal motor patterns. Tactile stimulation of one forewing of a locust elicits characteristic grooming movements of the ipsilateral hind leg. An initial targeted trajectory that moves the tarsus toward the site of stimulation is followed by a cyclic trajectory in the region of the stimulus. We have analyzed both components of this behavior to quantify the relative effects of somatotopic stimulus position and leg start posture on three parameters: initial movement direction, accuracy, and grooming distribution. Accuracy and grooming distribution were significantly affected by the stimulus location but were not influenced by the initial leg posture. Both cues systematically shifted the initial movement direction from the onset of the response. The subsequent cyclic component of grooming movements forms a behavioral continuum with no clustering in joint angle space. We therefore conclude that forewing grooming in locusts is generated by a single movement pattern that is continuously shifted by a sensory cue signaling position on the forewing surface. Both vertebrates and invertebrates can switch between distinct movement forms to groom different parts of their bodies. Our data provide the first evidence that invertebrates, like vertebrates, also have graded control of limb targeting within the somatosensory receptive field of a single form of motor response. PMID- 12773500 TI - Temporally irregular mnemonic persistent activity in prefrontal neurons of monkeys during a delayed response task. AB - An important question in neuroscience is whether and how temporal patterns and fluctuations in neuronal spike trains contribute to information processing in the cortex. We have addressed this issue in the memory-related circuits of the prefrontal cortex by analyzing spike trains from a database of 229 neurons recorded in the dorsolateral prefrontal cortex of 4 macaque monkeys during the performance of an oculomotor delayed-response task. For each task epoch, we have estimated their power spectrum together with interspike interval histograms and autocorrelograms. We find that 1). the properties of most (about 60%) neurons approximated the characteristics of a Poisson process. For about 25% of cells, with characteristics typical of interneurons, the power spectrum showed a trough at low frequencies (<20 Hz) and the autocorrelogram a dip near zero time lag. About 15% of neurons had a peak at <20 Hz in the power spectrum, associated with the burstiness of the spike train; 2). a small but significant task dependency of spike-train temporal structure: delay responses to preferred locations were characterized not only by elevated firing, but also by suppressed power at low (<20 Hz) frequencies; and 3). the variability of interspike intervals is typically higher during the mnemonic delay period than during the fixation period, regardless of the remembered cue. The high irregularity of neural persistent activity during the delay period is likely to be a characteristic signature of recurrent prefrontal network dynamics underlying working memory. PMID- 12773501 TI - Decisions on life and death: FOXO Forkhead transcription factors are in command when PKB/Akt is off duty. AB - Forkhead transcription factors of the FOXO family are important downstream targets of protein kinase B (PKB)/Akt, a kinase shown to play a decisive role in cell proliferation and cell survival. Direct phosphorylation by PKB/Akt inhibits transcriptional activation by FOXO factors, causing their displacement from the nucleus into the cytoplasm. Work from recent years has shown that this family of transcription factors regulates the expression of a number of genes that are crucial for the proliferative status of a cell, as well as a number of genes involved in programmed cell death. As such, these transcription factors appear to play an essential role in many of the effects of PKB/Akt on cell proliferation and survival. Indeed, in cells of the hematopoietic system, mere activation of a FOXO factor is sufficient to activate a variety of proapoptotic genes and to trigger apoptosis. In contrast, in most other cell types, activation of FOXO blocks cellular proliferation and drives cells into a quiescent state. In such cell types, FOXO factors also provide the protective mechanisms that are required to adapt to the altered metabolic state of quiescent cells. Thus, as PKB/Akt signaling is switched off, FOXO factors take over to determine the fate of a cell, long-term survival in a quiescent state, or programmed cell death. This review summarizes our current understanding of the mechanisms by which PKB/Akt and FOXO factors regulate these decisions. PMID- 12773502 TI - Nondisposable materials, chronic inflammation, and adjuvant action. AB - Why inflammatory responses become chronic and how adjuvants work remain unanswered. Macrophage-lineage cells are key components of chronic inflammatory reactions and in the actions of immunologic adjuvants. One explanation for the increased numbers of macrophages long term at sites of chronic inflammation could be enhanced cell survival or even local proliferation. The evidence supporting a unifying hypothesis for one way in which this macrophage survival and proliferation may be promoted is presented. Many materials, often particulate, of which macrophages have difficulty disposing, can promote monocyte/macrophage survival and even proliferation. Materials active in this regard and which can initiate chronic inflammatory reactions include oxidized low-density lipoprotein, inflammatory microcrystals (calcium phosphate, monosodium urate, talc, calcium pyrophosphate), amyloidogenic peptides (amyloid beta and prion protein), and joint implant biomaterials. Additional, similar materials, which have been shown to have adjuvant activity (alum, oil-in-water emulsions, heat-killed bacteria, CpG oligonucleotides, methylated bovine serum albumin, silica), induce similar responses. Cell proliferation can be striking, following uptake of some of the materials, when macrophage-colony stimulating factor is included at low concentrations, which normally promote mainly survival. It is proposed that if such responses were occurring in vivo, there would be a shift in the normal balance between cell survival and cell death, which maintains steady-state, macrophage-lineage numbers in tissues. Thus, there would be more cells in an inflammatory lesion or at a site of adjuvant action with the potential, following activation and/or differentiation, to perpetuate inflammatory or antigen specific, immune responses, respectively. PMID- 12773504 TI - Severe meningococcal disease is characterized by early neutrophil but not platelet activation and increased formation and consumption of platelet neutrophil complexes. AB - Approximately 25% of polymorphonuclear leukocytes (PMNL) circulate in heterotypic complexes with one or more activated platelets. These platelet-neutrophil complexes (PNC) require platelet CD62P expression for their formation and represent activated subpopulations of both cell types. In this study, we have investigated the presence, time course, and mechanisms of PNC formation in 32 cases of severe pediatric meningococcal disease (MD) requiring intensive care. There were marked early increases in PMNL CD11b/CD18 expression and activation, and reduced CD62L expression compared with intensive care unit control cases. Minimal platelet expression of the active form of alphaIIbbeta3 (GpIIb/IIIa) was seen. PNC were reduced on presentation and fell to very low levels after 24 h. Immunostaining of skin biopsies demonstrated that PNC appear outside the circulation in MD. In vitro studies of anticoagulated whole blood inoculated with Neisseria meningitidis supported these clinical findings with marked increases in PMNL CD11b/CD18 expression and activation but no detectable changes in platelet activated alphaIIbbeta3 or CD62P expression. In vitro PMNL activation with N. meningitidis (or other agonists) potentiated the formation of PNC in response to platelet activation with adenine diphosphate. Therefore, in severe MD, PMNL activation is likely to promote PNC formation, and we suggest that the reduced levels of PNC seen in established MD reflect rapid loss of PNC from the circulation rather than reduced formation. PMID- 12773503 TI - Essential involvement of IL-6 in the skin wound-healing process as evidenced by delayed wound healing in IL-6-deficient mice. AB - To clarify interleukin (IL)-6 roles in wound healing, we prepared skin excisions in wild-type (WT) and IL-6-deficient BALB/c [knockout (KO)] mice. In WT mice, the wound area was reduced to 50% of original size at 6 days after injury. Microscopically, leukocyte infiltration was evident at wound sites. Furthermore, the re-epithelialization rate was approximately 80% at 6 days after injury with increases in angiogenesis and hydroxyproline contents. The gene expression of IL 1, chemokines, adhesion molecules, transforming growth factor-beta1, and vascular endothelial growth factor was enhanced at the wound sites. In contrast, the enhanced expression of these genes was significantly reduced in KO mice. Moreover, in KO mice, the reduction of wound area was delayed with attenuated leukocyte infiltration, re-epithelialization, angiogenesis, and collagen accumulation. Finally, the administration of a neutralizing anti-IL-6 monoclonal antibody significantly delayed wound closure in WT mice. These observations suggest that IL-6 has crucial roles in wound healing, probably by regulating leukocyte infiltration, angiogenesis, and collagen accumulation. PMID- 12773505 TI - Ly49E expression points toward overlapping, but distinct, natural killer (NK) cell differentiation kinetics and potential of fetal versus adult lymphoid progenitors. AB - Using a new antibody, we found previously that contrary to adult natural killer (NK) cells, fetal NK cells have a unique phenotype, as they exclusively express Ly49E. This can be explained by an intrinsic different NK differentiation potential of fetal versus adult lymphoid progenitors, by immaturity of fetal NK cells or by instability of Ly49E expression. Here, we show that adult progenitor cells were still capable of differentiating into Ly49E-expressing NK cells but at a much lower frequency. Surprisingly, Ly49E expression in vitro did not require stromal cells. Kinetic analysis in vivo showed that Ly49E was expressed early, together with CD94/NKG2 and Ly49G2, followed by Ly49C, and finally Ly49D. Transfer of sorted Ly49E-positive fetal NK cells showed stable Ly49E expression, and later, part of these cells up-regulated other Ly49 members. These data indicate that although there are intrinsic differences, there is no strict fetal and adult wave of NK cell differentiation. PMID- 12773506 TI - 5-Lipoxygenase knockout mice exhibit a resistance to pleurisy and lung injury caused by carrageenan. AB - In the present study, by comparing the responses in wild-type (WT) mice and mice lacking [knockout (KO)] the 5-lipoxygenase (5-LO), we investigated the role played by 5-LO in the development of acute inflammation. When compared with carragenan-treated 5-LOWT mice, 5-LOKO mice, which had received carrageenan, exhibited a reduced degree of pleural exudation, polymorphonuclear cell migration. Lung myeloperoxidase activity, an index of neutrophil infiltration, was significantly reduced in 5-LOKO mice in comparison with 5-LOWT. Lung-tissue sections from carrageenan-treated 5-LOWT mice showed positive staining for intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), P-selectin, and E-selectin, which were mainly localized around vessels. The intensity and degree of ICAM-1, VCAM-1, P-selectin, and E-selectin were markedly reduced in tissue section from carrageenan-5-LOKO mice, which also improved the histological status of the inflamed lungs. Taken together, our results clearly demonstrate that 5-LO modulates neutrophil infiltration in the acute lung inflammation. PMID- 12773507 TI - A role for CD1d-restricted NKT cells in injury-associated T cell suppression. AB - Natural killer T (NKT) cells are known to modulate T cell responses during autoimmunity, tolerance, and antitumor immunity; however, their potential role in regulating the immune response to injury has not been reported. Using a murine model of burn injury, we investigated whether CD1d-restricted NKT cells played a role in the T cell suppression that occurs early after injury. A functional role for CD1d stimulation of NKT cells in the injury-related immune suppression was demonstrated by experiments in which the suppression of antigen (Ag)-specific delayed-type hypersensitivity and in vitro T cell-proliferative responses were prevented if mice were given anti-CD1d monoclonal antibody (mAb) systemically just before injury. The CD1d-NKT cell-dependent suppression of the T cell response after injury occurred in the absence of quantitative changes in NKT cells themselves or CD1d(+) Ag-presenting cells. We observed that elevated production of the immunosuppressive cytokine interleukin (IL)-4 correlated with burn-induced immune dysfunction, and we found that NKT cells but not conventional T cells were the source of IL-4 early after injury. Lastly, we observed that the injury-induced production of NKT cell-derived IL-4 could be blocked by systemic treatment of burn-injured mice with anti-CD1d mAb. Together, our results reveal a novel mechanism involving CD1d stimulation of NKT cells in the onset of T cell suppression that occurs subsequent to injury. PMID- 12773508 TI - Prostaglandin E2 modulates dendritic cell function via EP2 and EP4 receptor subtypes. AB - We have reported previously that PGE(2) inhibits dendritic cells (DC) functions. Because E prostanoid receptor (EPR) subtypes involved in this action are unknown, expression and functions of these receptors were examined in DC. Western blot and flow cytometry analyses showed that all EPRs were coexpressed in DC. In a dose dependent manner, lipopolysaccharide (LPS) enhanced EP(2)R/EP(4)R but not EP(1)R/EP(3)R expressions. NS-398, a cyclooxygenase (COX)-2-selective inhibitor, suppressed LPS-enhanced EP(2)R/EP(4)R expression, suggesting that COX-2-issued prostaglandin E(2) (PGE(2)) modulates DC function through stimulation of specific EPR subtypes. Using selective agonists, we found that butaprost, an EP(2)R agonist, and PGE(1) alcohol, an EP(2)R and EP(2)R/EP(4)R agonist, inhibited major histocompatibility complex class II expression and enhanced interleukin-10 production from DC. However, no effect was observed with sulprostone and 17 phenyl-omega-trinor-PGE(2), selective agonists for EP(1)R and EP(1)R/EP(3)R, respectively. Treatment of DC with dibutyryl cyclic adenosine monophosphate (cAMP), an analog of cAMP, mimics PGE(2)-induced, inhibitory effects. Taken together, our data demonstrate that EP(2)R/EP(4)R are efficient for mediating PGE(2)-induced modulation of DC functions. PMID- 12773509 TI - IgD-receptor (IgD-R) cross-linking partially protects murine T cells from dexamethasone-induced apoptosis. AB - Based on our previous findings that immunoglobulin D (IgD) receptor (IgD-R) cross linking with oligomeric IgD (IgD-R-xL) led to T cell activation, we examined the effect of IgD-R-xL on the expression of Fas antigen and apoptosis induction. In splenic T cells, IgD-R-xL followed by dexamethasone (dex) treatment resulted in a decreased percentage of Fas-positive cells as well as a decreased mean fluorescence intensity (P<0.05) when compared with cells treated with dex alone. There are significant differences in annexin-fluorescein isothiocyanate (FITC) and phosphatidylinositol (PI) staining between samples treated with dex alone and IgD-R-xL followed by dex-treated samples (P<0.05), suggesting a protective role for IgD-R-xL. No significant differences are seen in Fas antigen expression, annexin-FITC staining, and/or PI staining in murine T hybridoma (7C5) cells cultured under similar conditions (P<0.07). We hypothesize that ligation of IgD-R may predispose antigen-specific T lymphocytes for survival during primary immune responses when IgD-positive B cells serve as antigen-presenting cells. PMID- 12773510 TI - Antibody-mediated blockade of the CXCR3 chemokine receptor results in diminished recruitment of T helper 1 cells into sites of inflammation. AB - Naive T cells, when activated by specific antigen and cytokines, up-regulate adhesion molecules as well as chemokine receptors on their surface, which allows them to migrate to inflamed tissues. Human studies have shown that CXCR3 is one of the chemokine receptors that is induced during T cell activation. Moreover, CXCR3-positive T cells are enriched at inflammatory sites in patients with autoimmune diseases such as rheumatoid arthritis and multiple sclerosis. In this study, we use a mouse model of inflammation to demonstrate that CXCR3 is required for activated T cell transmigration to inflamed tissue. Using an anti- mCXCR3 antibody, we have shown that in vitro-differentiated T helper (Th) 1 and Th2 cells up-regulated CXCR3 upon stimulation with specific antigen/major histocompatibility complex. However, only Th1 cells, when adoptively transferred to syngeneic recipients, are efficiently recruited to the peritoneum in an adjuvant-induced peritonitis model. Furthermore, the neutralizing anti-mCXCR3 antibody profoundly inhibits the recruitment of Th1 cells to the inflamed peritoneum. Real-time, quantitative reverse transcriptase-polymerase chain reaction analysis demonstrates that the CXCR3 ligands, interferon (IFN)-inducible protein 10 (CXCL10) and IFN-inducible T cell alpha chemoattractant (CXCL11), are among the many chemokines induced in the adjuvant-treated peritoneum. The anti mCXCR3 antibody is also effective in inhibiting a delayed-type hypersensitivity response, which is largely mediated by enhanced trafficking of activated T cells to peripheral inflammatory sites. Collectively, our results suggest that CXCR3 has a critical role in T cell transmigration to sites of inflammation and thus, may serve as a molecular target for anti-inflammatory therapies. PMID- 12773512 TI - Role of activin A in murine mast cells: modulation of cell growth, differentiation, and migration. AB - Activins, members of the transforming growth factor-beta (TGF-beta) superfamily, are potent growth and differentiation factors. Our previous studies revealed that activin A, a homodimer of inhibin/activin beta(A), was induced in mast cells and peritoneal macrophages in response to their activation. In the present study, we examined the roles of activin A in murine bone marrow-derived, cultured mast cell progenitors (BMCMCs), which expressed gene transcripts for molecules involved in activin signaling, suggesting that BMCMCs could be target cells of activin A. Treatment of activin A inhibited 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide uptake into BMCMCs in a dose-dependent manner. The IC(50) concentration was 2.1 nM, which was less potent than 185 pM TGF-beta(1). Activin A treatment caused morphological changes toward the differentiated cells at 2 nM and up-regulated mRNA of mouse mast cell protease-1 (mMCP-1), a marker enzyme of mature mucosal mast cells, at 1 nM. Activin A also showed activity in inducing migration of BMCMCs; the optimal concentration for maximal migration was 10 pM, which was much lower than the concentrations to inhibit cell growth and to activate the mMCP-1 gene. Taking the present results together with our previous results, it is suggested that activin A secreted from activated immune cells recruits mast cell progenitors to sites of inflammation and that with increasing activin A concentration, the progenitors differentiate into mature mast cells. Thus, activin A may positively regulate the functions of mast cells as effector cells of the immune system. PMID- 12773511 TI - Identification of a novel CpG DNA class and motif that optimally stimulate B cell and plasmacytoid dendritic cell functions. AB - Recent reports have identified two major classes of CpG motif-containing oligodeoxynucleotide immunostimulatory sequences (ISS): uniformly modified phosphorothioate (PS) oligodeoxyribonucleotides (ODNs), which initiate B cell functions but poorly activate dendritic cells (DCs) to make interferon (IFN) alpha, and chimeric PS/phosphodiester (PO) ODNs containing runs of six contiguous guanosines, which induce very high levels of plasmacytoid DC (PDC)-derived IFN alpha but poorly stimulate B cells. We have generated the first reported ISS, C274, which exhibits very potent effects on all human immune cells known to recognize ISS. C274 is a potent inducer of IFN-gamma/IFN-alpha from peripheral blood mononuclear cells and exhibits accelerated kinetics of activity compared with standard ISS. This ODN also effectively stimulates B cells to proliferate, secrete cytokines, and express costimulatory antigens. In addition, C274 specifically activates PDCs to undergo maturation and secrete cytokines, including very high levels of IFN-alpha. Sequence variation studies based on C274 were used to identify the general motif requirements for this novel and distinct class of ISS. In contrast, chimeric PO/PS CpG-containing ODNs with polyguanosine sequences exert a differential pattern of ISS activity compared with C274, perhaps in part as a result of their greatly different structural nature. This pattern is composed of high IFN-alpha/IFN-gamma induction and low DC maturation in the absence of B cell stimulation. In conclusion, we have generated a novel class of ISS that transcends the limitations ascribed to classes described previously in that it provides excellent stimulation of B cells and simultaneously activates PDCs to differentiate and secrete large amounts of type I IFN. PMID- 12773513 TI - Activation of CR3-mediated phagocytosis by MSP requires the RON receptor, tyrosine kinase activity, phosphatidylinositol 3-kinase, and protein kinase C zeta. AB - Macrophage-stimulating protein (MSP) promotes the phagocytosis of C3bi-coated erythrocytes by resident peritoneal macrophages, although the mechanism by which this occurs is largely unknown. We show that MSP-induced complement-mediated phagocytosis requires the RON receptor tyrosine kinase and the alphaMbeta2 integrin, as evidenced by the inability of RON-/- and alphaM-/- peritoneal macrophages to augment phagocytosis of complement-coated sheep erythrocytes in response to MSP. MSP stimulation of macrophages results in tyrosine phosphorylation and AKT activation, and inhibitor studies demonstrate a phagocytic requirement for tyrosine kinase and phosphatidylinositol 3-kinase (PI 3K) activity as well as activity of the atypical protein kinase C (PKC) isoform zeta, which localizes to MSP-induced phagosomes containing complement-coated beads. Additionally, MSP augments the ability of peritoneal macrophages to bind to intercellular adhesion molecule-1 (ICAM-1) via the alphaMbeta2 integrin. MSP induced ICAM-1 adhesion is also dependent on tyrosine kinase activity, PI-3K, and PKC zeta, indicating that these signaling requirements are upstream of complement receptor 3 activation. PMID- 12773514 TI - Up-regulated expression of MICA on activated T lymphocytes involves Lck and Fyn kinases and signaling through MEK1/ERK, p38 MAP kinase, and calcineurin. AB - Major histocompatibility complex class I-related chain (MICA) is a cell stress regulated molecule recognized by cytotoxic cells expressing the NKG2D molecule. MICA can be induced on T cells after CD3 or CD28 engagement. Here, we investigated the intracellular pathways leading to activation-induced expression of MICA. The Src kinase inhibitor PP1 inhibited up-regulated expression of MICA on anti-CD3-stimulated T cells. Downstream signaling routes involved mitogen activated protein kinase (MAPK) kinase (MEK)1/extracellular signal-regulated kinase (ERK), p38 MAPK, and calcineurin, as MICA expression was prevented by U0126, SB202190, cyclosporin A, and FK506. Also, Lck and Fyn as well as MEK1/ERK and p38 MAPK were found to regulate MICA expression in anti-CD28/phorbol 12 myristate 13-acetate-stimulated T cells. Expression of MICA on activated T cells involved interleukin-2-dependent signaling routes triggered by Janus tyrosine kinases/signal transducer and activators of transcription and p70(S)(6) kinase, as it could be inhibited by AG490 and rapamycin. This is the first demonstration of the intracellular pathways involved in activation-induced expression of MICA, which may reveal potential targets for immune intervention to modulate MICA expression in pathological disorders. PMID- 12773515 TI - The effect of phosphatases SHP-1 and SHIP-1 on signaling by the ITIM- and ITAM containing Fcgamma receptors FcgammaRIIB and FcgammaRIIA. AB - Inositol and tyrosine phosphatases have been implicated in inhibitory signaling by an Fc receptor for immunoglobulin G, FcgammaRIIB, in B cells, mast cells, and monocytes. Here, we propose a role for the Src homology 2 (SH2)-containing tyrosine phosphatase-1 (SHP-1) in FcgammaRIIB-mediated inhibition of FcgammaR signaling. Coexpression of SHP-1 enhances FcgammaRIIB-mediated inhibition of FcgammaRIIA phagocytosis in COS-1 cells. SHP-1 also enhances the reduction in FcgammaRIIA tyrosine phosphorylation that accompanies this inhibition. Significantly, tyrosine phosphorylation of Syk kinase is substantially inhibited by SHP-1. Furthermore, the activation of SHP-1 tyrosine phosphorylation is observed following stimulation of FcgammaRII in COS-1 cells and in human monocytes. The SH2 domain containing inositol phosphatase (SHIP), SHIP-1 also enhances FcgammaRIIB-mediated inhibition of FcgammaRIIA, indicating that FcgammaRIIB can use more than one pathway for its inhibitory action. In addition, SHP-1 and SHIP-1 can inhibit FcgammaRIIA phagocytosis and signal transduction in the absence of FcgammaRIIB. The data support emerging evidence that SH2 containing phosphatases, such as SHP-1 and SHIP-1, can modulate signaling by "activating" receptors. PMID- 12773516 TI - Role of Rho-family GTPase Cdc42 in polarized expression of lymphocyte appendages. AB - Lymphocytes polarize for motility by developing a broad anterior, where lamellipodia arise, and a simple stalk-like posterior appendage, the uropod. Through time-lapse analysis of normal and leukemic human T cells, it was found that this polarized form is maintained by a mechanism that excludes lamellipodia from the uropod. Lamellipodia regularly traveled rearward to encroach upon the uropod but disassembled abruptly at the uropod border. This exclusion of lamellipodia from the uropod required the Rho-family guanosine triphosphatase Cdc42. Reduction of Cdc42 activity by expression of dominant-negative Cdc42 resulted in "two headed" cells in which lamellipodia persisted at the distal end of the uropod. Random and chemotactic motility were impaired. Increased Cdc42 activity, induced by expression of activated, mutant Cdc42, was accompanied by a general loss of lamellipodia. The results suggest that one role of Cdc42 in lymphocyte motility is to preserve polarity by concentrating lamellipodial disassembly signals in the uropod. PMID- 12773517 TI - Expression of myeloperoxidase (MPO) by neutrophils is necessary for their activation by anti-neutrophil cytoplasm autoantibodies (ANCA) against MPO. AB - Anti-neutrophil cytoplasm autoantibodies (ANCA) directed against proteinase-3 and myeloperoxidase (MPO) activate tumor necrosis factor-alpha-primed neutrophils in vitro. We used neutrophils from one completely and one partially MPO-deficient donor to assess the requirement of MPO expression for neutrophil activation by anti-MPO antibodies. The MPO deficiencies were defined enzymatically, by immunocytochemistry and by immunoblotting. The mutations in the MPO genes of these donors were identified as a combination of a novel splice-site mutation at the 3' end of intron 11 (A-2-->C), a deletion of 14 nucleotides in exon 9 (A1555 C1568), and a novel C1907 --> T (636Thr-->Met) substitution in exon 11 in the completely MPO-deficient donor and as the same splice-site mutation and a novel C995 --> T (332Ala-->Val) substitution in exon 7 in the partially MPO-deficient donor. Monoclonal antibody 4.15 against MPO and MPO-ANCA-immunoglobulin G induced no superoxide anion production in these MPO-deficient neutrophils despite a normal production induced by other stimuli. Thus, the presence of MPO is a conditio sine qua non for neutrophil activation by anti-MPO antibodies. Moreover, we demonstrated that by means of these MPO-deficient cells, hydrogen peroxide may diffuse from neutrophils to surrounding cells, which may contribute to the damage induced by oxygen radicals in the pathology of systemic vasculitides. PMID- 12773519 TI - Histone acetylation and chromatin conformation are regulated separately at the TNF-alpha promoter in monocytes and macrophages. AB - Tumor necrosis factor alpha (TNF-alpha) is a proinflammatory cytokine, which participates in a wide range of immunoregulatory activities. It is generally produced at highest levels by cells of the myeloid lineage in response to activation of pathogen recognition receptors such as Toll-like receptors. Impaired production predisposes to infection with intracellular organisms, and overproduction results in systemic or organ-specific inflammation. Control of expression is essential to maintain homeostasis, and this control is mediated via multiple strategies. We examined two separate aspects of chromatin accessibility in this study of the human TNF-alpha promoter. We examined the role of histone acetylation and chromatin remodeling in cell lines and primary cells and identified two individual steps associated with activation of TNF-alpha production. Histone H3 and H4 acetylation was found to be strongly dependent on the developmental stage of human monocytes. It did not appear to be regulated by acute stimuli, and instead, chromatin remodeling was found to occur after acute stimuli in a cell line competent to produce TNF-alpha. These data suggest that there is a hierarchy of controls regulating expression of TNF-alpha. Acetylation of histones is a prerequisite but is insufficient on its own for TNF-alpha production. PMID- 12773518 TI - p53 deficiency and defective mitotic checkpoint in proliferating T lymphocytes increase chromosomal instability through aberrant exit from mitotic arrest. AB - During the proliferation of T cells for successful immune responses against pathogens, the fine regulation of cell cycle is important to the maintenance of T cell homeostasis and the prevention of lymphoproliferative disorders. However, it remains to be elucidated how the cell cycle is controlled at the mitotic phase in proliferating T cells. Here, we show that during the proliferation of primary T cells, the disruption of the mitotic spindle leads to cell-cycle arrest at mitosis and that prolonged mitotic arrest results in not only apoptosis but also the form of chromosomal instability observed in human cancers. It is interesting that in response to spindle damage, the phosphorylation of BubR1, a mitotic checkpoint kinase, was significantly induced in proliferating T cells, and the expression of the dominant-negative mutant of BubR1 compromised mitotic arrest and subsequent apoptosis and thus led to the augmentation of polyploidy formation. We also show that in response to prolonged spindle damage, the expression of p53 but not of p73 was significantly induced. In addition, following sustained mitotic arrest, p53-deficient T cells were found to be more susceptible to polyploidy formation than the wild type. These results suggest that during flourishing immune response, mitotic checkpoint and p53 play important roles in the prevention of chromosomal instability and in the maintenance of the genomic integrity of proliferating T cells. PMID- 12773520 TI - Environmentally-induced changes in protein composition in developing grains of wheat are related to changes in total protein content. AB - Nitrogen (N) nutrition, post-anthesis temperature and drought-induced changes in the kinetics of accumulation of dry mass, total grain N and protein fractions (albumins-globulins, amphiphils, gliadins, and glutenins) contents were examined for winter wheat (Triticum aestivum L.). Crops were grown in controlled environment tunnels in 1994 and 1998. In 1994, five post-anthesis temperatures averaging from 15-25 degrees C were applied during grain-filling. In 1998 two post-anthesis temperatures averaging 13 degrees C and 20 degrees C were applied and factorized with two post-anthesis water regimes. In 1994 crops also were grown in the field, where different application rates and timing of N nutrition were tested. When expressed in thermal time, the kinetics of accumulation of the protein fractions were not significantly affected by post-anthesis temperature or drought; whereas N nutrition significantly increased the rate and duration of accumulation of storage proteins. Albumin-globulin proteins accumulated during the early stage of grain development. The rate of accumulation of that fraction decreased significantly at c. 250 degrees Cd after anthesis, when the storage proteins (gliadins and glutenins) started to accumulate significantly. Single allometric relationships for the different environmental conditions exist between the quantity of each protein fraction and the total quantity of N per grain. From these results it was concluded that (1) the process of N partitioning is neither significantly affected by post-anthesis temperature or drought nor by the rate and timing of N nutrition and (2) at maturity, variations in protein fraction composition are mainly because of differences in the total quantity of N accumulated during grain-filling. PMID- 12773521 TI - The responses of guard and mesophyll cell photosynthesis to CO2, O2, light, and water stress in a range of species are similar. AB - High resolution chlorophyll a fluorescence imaging was used to compare the photosynthetic efficiency of PSII electron transport (estimated by Fq'/Fm') in guard cell chloroplasts and the underlying mesophyll in intact leaves of six different species: Commelina communis, Vicia faba, Amaranthus caudatus, Polypodium vulgare, Nicotiana tabacum, and Tradescantia albifora. While photosynthetic efficiency varied between the species, the efficiencies of guard cells and mesophyll cells were always closely matched. As measurement light intensity was increased, guard cells from the lower leaf surfaces of C. communis and V. faba showed larger reductions in photosynthetic efficiency than those from the upper surfaces. In these two species, guard cell photosynthetic efficiency responded similarly to that of the mesophyll when either light intensity or CO2 concentration during either measurement or growth was changed. In all six species, reducing the O2 concentration from 21% to 2% reduced guard cell photosynthetic efficiency, even for the C4 species A. caudatus, although the mesophyll of the C4 species did not show any O2 modulation of photosynthetic efficiency. This suggests that Rubisco activity is significant in the guard cells of these six species. When C. communis plants were water-stressed, the guard cell photosynthetic efficiency declined in parallel with that of the mesophyll. It was concluded that the photosynthetic efficiency in guard cells is determined by the same factors that determine it in the mesophyll. PMID- 12773522 TI - 12CO2 emission from different metabolic pathways measured in illuminated and darkened C3 and C4 leaves at low, atmospheric and elevated CO2 concentration. AB - The detection of 12CO2 emission from leaves in air containing 13CO2 allows simple and fast determination of the CO2 emitted by different sources, which are separated on the basis of their labelling velocity. This technique was exploited to investigate the controversial effect of CO2 concentration on mitochondrial respiration. The 12CO2 emission was measured in illuminated and darkened leaves of one C4 plant and three C3 plants maintained at low (30-50 ppm), atmospheric (350-400 ppm) and elevated (700-800 ppm) CO2 concentration. In C3 leaves, the 12CO2 emission in the light (Rd) was low at ambient CO2 and was further quenched in elevated CO2, when it was often only 20-30% of the 12CO2 emission in the dark, interpreted as the mitochondrial respiration in the dark (Rn). Rn was also reduced in elevated CO2. At low CO2, Rd was often 70-80% of Rn, and a burst of 12CO2 was observed on darkening leaves of Mentha sativa and Phragmites australis after exposure for 4 min to 13CO2 in the light. The burst was partially removed at low oxygen and was never observed in C4 leaves, suggesting that it may be caused by incomplete labelling of the photorespiratory pool at low CO2. This pool may be low in sclerophyllous leaves, as in Quercus ilex where no burst was observed. Rd was inversely associated with photosynthesis, suggesting that the Rd/Rn ratio reflects the refixation of respiratory CO2 by photosynthesizing leaves rather than the inhibition of mitochondrial respiration in the light, and that CO2 produced by mitochondrial respiration in the light is mostly emitted at low CO2, and mostly refixed at elevated CO2. In the leaves of the C4 species Zea mays, the 12CO2 emission in the light also remained low at low CO2, suggesting efficient CO2 refixation associated with sustained photosynthesis in non photorespiratory conditions. However, Rn was inhibited in CO2-free air, and the velocity of 12CO2 emission after darkening was inversely associated with the CO2 concentration. The emission may be modulated by the presence of post-illumination CO2 uptake deriving from temporary imbalance between C3 and C4 metabolism. These experiments suggest that this uptake lasts longer at low CO2 and that the imbalance is persistent once it has been generated by exposure to low CO2. PMID- 12773523 TI - Al-induced efflux of organic acid anions is poorly associated with internal organic acid metabolism in triticale roots. AB - The secretion of organic acid anions from roots has been identified as a mechanism of resistance to Al. However, the process leading to the secretion of organic acid anions is poorly understood. The effect of Al on organic acid metabolism was investigated in two lines of triticale (xTriticosecale Wittmark) differing in Al-induced secretion of malate and citrate and in Al resistance. The site of Al-induced secretion of citrate and malate from a resistant line was localized to the root apices (terminal 5 mm). The levels of citrate (root apices and mature root segments) and malate (mature segments only) in roots increased during exposure to Al, but similar changes were observed in both triticale genotypes. The in vitro activities of four enzymes involved in malate and citrate metabolism (citrate synthase, phosphoenolpyruvate carboxylase, malate dehydrogenase, and NADP-isocitrate dehydrogenase) were similar for sensitive and resistant lines in both root apices and mature root segments. The response of these enzymes to pH did not differ between tolerant and sensitive lines or in the presence and absence of Al. Moreover, cytoplasmic and vacuolar pH were not affected by exposure to Al in either line. Together, these results indicate that the Al-dependent efflux of organic acid anions from the roots of triticale is not regulated by their internal levels in the roots or by the capacity of the root cells to synthesize malate and citrate. PMID- 12773524 TI - Nitrate-independent expression of plant nitrate reductase in Lotus japonicus root nodules. AB - Nitrate-independent nitrate reductase (NR) activity is generally found in legume root nodules. Therefore, the effects of nitrate on plant NR activity and mRNA were investigated in the root nodules of Lotus japonicus (L. japonicus). Both NR activity and mRNA levels in roots and root nodules were up-regulated by the addition of nitrate. In the absence of nitrate, NR activity and mRNA were detected in root nodules but not in roots. Southern blotting analysis indicates that NR is encoded by a single gene in L. japonicus. No nitrate was detected in the root nodules or roots of plants grown in the absence of nitrate, while its accumulation was observed in plants supplied with exogenous nitrate. These results indicate that inducible-type NR can be expressed in root nodules in the absence of nitrate. The activation state of the nitrate-independent activity of NR was as high as that of NR activity induced by nitrate. NR mRNA expressed independently of nitrate in root nodules without nitrate was localized in the infected regions of the root nodules. Thus, the expression could be related to the specific structure and environment of root nodules. PMID- 12773525 TI - Sugar uptake and proton release by protoplasts from the infected zone of Vicia faba L. nodules: evidence against apoplastic sugar supply of infected cells. AB - Symbiotic dinitrogen fixation of legume nodules is fuelled by phloem-imported carbohydrates. These have to pass several cell layers to reach cells infected with Rhizobium bacteroids. It is unclear whether apoplastic steps are involved in carbohyd-rate translocation within the nodule. Protoplasts were isolated from the infected and uninfected cells of the central tissue of Vicia faba nodules using a recently developed protocol. These protoplasts were used to elucidate pathways for sugar transport in this tissue. Both types of protoplasts released protons into the medium. Acidification was inhibited by vanadate and erythrosin B. However, it was stimulated by fusicoccin only in uninfected cells. A symport of sugars with protons can therefore be energized in both cell types. Uptake of 14C labelled sugars was determined using a phthalate centrifugation technique. Uninfected protoplasts accumulated glucose through high-affinity H+/glucose symport that was not competitively inhibited by fructose or sucrose. Uninfected protoplasts also absorbed sucrose with biphasic kinetics. At 0.1, 1, and 10 mM sucrose, uptake was inhibited by CCCP. Fusicoccin did not stimulate the linear phase of sucrose uptake. Glucose inhibited sucrose uptake nearly completely. This was not related to sucrose cleavage in the medium because sucrose was absorbed at a much higher rate than glucose, and glucose concentration did not increase in sucrose-containing protoplast suspensions. By contrast with uninfected protoplasts, infected cells did not show transporter-mediated glucose or sucrose uptake. The findings underline a role of uninfected cells in sugar translocation. Infected cells are not apoplastically supplied with sugars and possibly depend on uninfected cells for carbon supply. PMID- 12773526 TI - Sialoside specificity of the siglec family assessed using novel multivalent probes: identification of potent inhibitors of myelin-associated glycoprotein. AB - Ten of the 11 known human siglecs or their murine orthologs have been evaluated for their specificity for over 25 synthetic sialosides representing most of the major sequences terminating carbohydrate groups of glycoproteins and glycolipids. Analysis has been performed using a novel multivalent platform comprising biotinylated sialosides bound to a streptavidin-alkaline phosphatase conjugate. Each siglec was found to have a unique specificity for binding 16 different sialoside-streptavidin-alkaline phosphatase probes. The relative affinities of monovalent sialosides were assessed for each siglec in competitive inhibition studies. The quantitative data obtained allows a detailed analysis of each siglec for the relative importance of sialic acid and the penultimate oligosaccharide sequence on binding affinity and specificity. Most remarkable was the finding that myelin-associated glycoprotein (Siglec-4) binds with 500-10,000-fold higher affinity to a series of mono- and di-sialylated derivatives of the O-linked T antigen (Galbeta(1-3)-GalNAc(alpha)OThr) as compared with alpha-methyl-NeuAc. PMID- 12773527 TI - Novel aldoxime dehydratase involved in carbon-nitrogen triple bond synthesis of Pseudomonas chlororaphis B23. Sequencing, gene expression, purification, and characterization. AB - Analysis of the nitrile hydratase gene cluster involved in nitrile metabolism of Pseudomonas chlororaphis B23 revealed that it contains one open reading frame encoding aldoxime dehydratase upstream of the amidase gene. The amino acid sequence deduced from this open reading frame shows similarity (32% identity) with that of Bacillus phenylacetaldoxime dehydratase (Kato, Y., Nakamura, K., Sakiyama, H., Mayhew, S. G., and Asano, Y. (2000) Biochemistry 39, 800-809). The gene product expressed in Escherichia coli catalyzed the dehydration of aldoxime into nitrile. The Pseudomonas aldoxime dehydratase (OxdA) was purified from the E. coli transformant and characterized. OxdA shows an absorption spectrum with a Soret peak that is characteristic of heme, demonstrating that it is a hemoprotein. For its activity, this enzyme required a reducing reagent, Na2S2O4, but did not require FMN, which is crucial for the Bacillus enzyme. The enzymatic reaction was found to be catalyzed when the heme iron of the enzyme was in the ferrous state. Calcium as well as iron was included in the enzyme. OxdA reduced by Na2S2O4 had a molecular mass of 76.2 kDa and consisted of two identical subunits. The kinetic parameters of OxdA indicated that aliphatic aldoximes are more effective substrates than aromatic aldoximes. A variety of spectral shifts in the absorption spectra of OxdA were observed upon the addition of each of various compounds (i.e. redox reagents and heme ligands). Moreover, the addition of the substrate to OxdA gave a peak that would be derived from the intermediate in the nitrile synthetic reaction. P. chlororaphis B23 grew and showed the OxdA activity when cultured in a medium containing aldoxime as the sole carbon and nitrogen source. Together with these findings, Western blotting analysis of the extracts using anti-OxdA antiserum revealed that OxdA is responsible for the metabolism of aldoxime in vivo in this strain. PMID- 12773528 TI - Domain truncation studies reveal that the streptokinase-plasmin activator complex utilizes long range protein-protein interactions with macromolecular substrate to maximize catalytic turnover. AB - To explore the interdomain co-operativity during human plasminogen (HPG) activation by streptokinase (SK), we expressed the cDNAs corresponding to each SK domain individually (alpha, beta, and gamma), and also their two-domain combinations, viz. alphabeta and betagamma in Escherichia coli. After purification, alpha and beta showed activator activities of approximately 0.4 and 0.05%, respectively, as compared with that of native SK, measured in the presence of human plasmin, but the bi-domain constructs alphabeta and betagamma showed much higher co-factor activities (3.5 and 0.7% of native SK, respectively). Resonant Mirror-based binding studies showed that the single-domain constructs had significantly lower affinities for "partner" HPG, whereas the affinities of the two-domain constructs were remarkably native-like with regards to both binary mode as well as ternary mode ("substrate") binding with HPG, suggesting that the vast difference in co-factor activity between the two- and three-domain structures did not arise merely from affinity differences between activator species and HPG. Remarkably, when the co-factor activities of the various constructs were measured with microplasminogen, the nearly 50-fold difference in the co-factor activity between the two- and three-domain SK constructs observed with full-length HPG as substrate was found to be dramatically attenuated, with all three types of constructs now exhibiting a low activity of approximately 1-2% compared to that of SK.HPN and HPG. Thus, the docking of substrate through the catalytic domain at the active site of SK-plasmin(ogen) is capable of engendering, at best, only a minimal level of co-factor activity in SK.HPN. Therefore, apart from conferring additional substrate affinity through kringle mediated interactions, reported earlier (Dhar et al., 2002; J. Biol. Chem. 277, 13257), selective interactions between all three domains of SK and the kringle domains of substrate vastly accelerate the plasminogen activation reaction to near native levels. PMID- 12773529 TI - Cell membrane lipid rafts mediate caveolar endocytosis of HIV-1 Tat fusion proteins. AB - The transactivator protein of human immunodeficiency virus type 1 Tat has the unique property of mediating the delivery of large protein cargoes into the cells when present in the extracellular milieu. Here we show that Tat fusion proteins are internalized by the cells through a temperature-dependent endocytic pathway that originates from cell membrane lipid rafts and follows caveolar endocytosis. These conclusions are supported by the study of the slow kinetics of the internalization of Tat endosomes, by their resistance to nonionic detergents, the colocalization of internalized Tat with markers of caveolar endocytosis, and the impairment of the internalization process by drugs that disrupt lipid rafts or disturb caveolar trafficking. These results are of interest for all those who exploit Tat as a vehicle for transcellular protein delivery. PMID- 12773531 TI - Apoptosome-independent pathway for apoptosis. Biochemical analysis of APAF-1 defects and biological outcomes. AB - Induction and execution of apoptosis programs are generally believed to be mediated through a hierarchy of caspase activation. By using two cellular variants obtained from the L1210 cell line (L1210/S and L1210/0), we have shown previously that staurosporine induces apoptotic cell death through both caspase dependent and caspase-independent pathways. Both pathways normally coexisted in L1210/S cells, whereas L1210/0 cells lacked the ability to activate caspases despite the confirmed presence of both procaspase-3 and -9. Here we show that this defect in caspase activation is not due to mechanisms such as an absence of cytochrome c release, the expression of non-functional caspases, or the presence of an endogenous inhibitor but results from the loss of apoptosis protease activator protein-1 (APAF-1) expression. This absence of APAF-1 protein results from multiple alterations at both genomic and transcriptional levels. However, although this lack of APAF-1 delays the apoptotic program, it does not hamper its execution. Importantly, in these cells, apoptosis develops not only in an APAF-1 independent way but also in the absence of caspase-3 and -9 activation. Altogether these findings provide evidence that apoptosis may occur through alternative signaling pathways independent of APAF-1 expression and totally dissociated from any caspase processing. Therefore, the L1210/0 variant sub-line provides a valuable tool for the elucidation of these pathways. PMID- 12773532 TI - ATP-induced reverse temperature effect in isohemoglobins from the endothermic porbeagle shark (Lamna nasus). AB - The evolutionary convergence of endothermic tunas and lamnid sharks is unique. Their heat exchanger-mediated endothermy represents an interesting example of the evolutionary pressure associated with this specific characteristic. To assess the implications of endothermy for gas transport and the possible contribution of hemoglobin (Hb), we investigated the effect of temperature on the oxygen equilibria of purified isohemoglobin components V and III from the porbeagle shark (Lamna nasus). In the absence of ATP the effect of temperature on oxygen affinity is normal in both Hb III (P50 = 0.9 and 2.2 torr at 10 and 26 degrees C, respectively) and Hb V (P50 = 1.5 and 2.5 torr at 10 and 26 degrees C, respectively). In the presence of this effector P50 decreases with increasing temperature in both components (P50 at 10 and 26 degrees C = 9.9 and 8.4 torr (Hb III), respectively, and 9.6 and 7.4 torr (Hb V), respectively. The reverse temperature effect in the presence of ATP will reduce the risk of oxygen loss from the arterial to the venous blood by lowering the oxygen tension gradient between the blood vessels. The mechanism behind the reverse temperature effect resembles that found in the bluefin tuna (Thunnus thynnus), an endothermic teleost, thus evidencing further convergent evolution. PMID- 12773530 TI - The functional integrity of the serpin domain of C1-inhibitor depends on the unique N-terminal domain, as revealed by a pathological mutant. AB - C1-inhibitor (C1-Inh) is a serine protease inhibitor (serpin) with a unique, non conserved N-terminal domain of unknown function. Genetic deficiency of C1-Inh causes hereditary angioedema. A novel type of mutation (Delta 3) in exon 3 of the C1-Inh gene, resulting in deletion of Asp62-Thr116 in this unique domain, was encountered in a hereditary angioedema pedigree. Because the domain is supposedly not essential for inhibitory activity, the unexpected loss-of-function of this deletion mutant was further investigated. The Delta 3 mutant and three additional mutants starting at Pro76, Gly98, and Ser115, lacking increasing parts of the N terminal domain, were produced recombinantly. C1-Inh76 and C1-Inh98 retained normal conformation and interaction kinetics with target proteases. In contrast, C1-Inh115 and Delta 3, which both lack the connection between the serpin and the non-serpin domain via two disulfide bridges, were completely non-functional because of a complex-like and multimeric conformation, as demonstrated by several criteria. The Delta 3 mutant also circulated in multimeric form in plasma from affected family members. The C1-Inh mutant reported here is unique in that deletion of an entire amino acid stretch from a domain not shared by other serpins leads to a loss-of-function. The deletion in the unique N-terminal domain results in a "multimerization phenotype" of C1-Inh, because of diminished stability of the central beta-sheet. This phenotype, as well as the location of the disulfide bridges between the serpin and the non-serpin domain of C1-Inh, suggests that the function of the N-terminal region may be similar to one of the effects of heparin in antithrombin III, maintenance of the metastable serpin conformation. PMID- 12773533 TI - The number of repeat sequences in microtubule-associated protein 4 affects the microtubule surface properties. AB - The microtubule-binding domain of MAP4, a ubiquitous microtubule-associated protein, contains a Repeat region with tandemly organized repeat sequences. In this study, we focused on the variations of the Repeat region, and searched for MAP4 isoforms with diverse Repeat region organizations. We successfully isolated four types of MAP4 cDNAs, which differed from each other in both the number and the arrangement of the repeat sequences, from a single source (bovine adrenal gland). To examine the functional differences among the isoforms, we prepared the microtubule-binding domain polypeptides of three of the four isoforms, and examined their activities. The isoform fragments showed similar degrees of microtubule assembly promoting activity and microtubule binding affinity. This result suggested that the Repeat region variation is not important for the control of microtubule dynamics, which is believed to be the main function of MAPs. On the other hand, the microtubule bundle-forming activity differed among the isoform fragments. The bundle formation was augmented by increasing the number of repeat sequences in the fragments. Based on these results, we propose the hypothesis that the role of the MAP4 isoforms is to regulate the surface charge of microtubules. PMID- 12773535 TI - Generation of a FasL-based proapoptotic fusion protein devoid of systemic toxicity due to cell-surface antigen-restricted Activation. AB - We describe the construction of a FasL fusion protein devoid of systemic toxicity, inducing apoptosis only on cell-surface antigen-positive cells. The fusion protein consists carboxyl-terminally of the extracellular domain of FasL and amino-terminally of a fibroblast activation protein (FAP)-specific single chain antibody fragment (sc40-FasL). The latter allows immobilization-dependent conversion of the inactive soluble FasL fusion protein into an entity with membrane FasL-like activity. Thus, sc40-FasL efficiently induced apoptosis only in FAP-expressing cells. In accordance with a strict target-selective activity of sc40-FasL, the intravenous application of this reagent in mice revealed no signs of systemic toxicity and prevented growth of xenotransplanted FAP-positive (but not FAP-negative) tumor cells. The principle described here for the first time, in which cell-surface antigen-mediated activation of Fas permits local activation of Fas in vivo, opens novel avenues for the use of Fas signaling in cancer therapy. PMID- 12773534 TI - 11,12-Epoxyeicosatrienoic acid-induced inhibition of FOXO factors promotes endothelial proliferation by down-regulating p27Kip1. AB - Cytochrome P450-derived epoxyeicosatrienoic acids (EETs) stimulate endothelial cell proliferation and angiogenesis. In this study, we investigated the involvement of the forkhead box, class O (FOXO) family of transcription factors and their downstream target p27Kip1 in EET-induced endothelial cell proliferation. Incubation of human umbilical vein endothelial cells with 11,12 EET induced a time- and dose-dependent decrease in p27Kip1 protein expression, whereas p21Cip1 was not significantly affected. This effect on p27Kip1 protein was associated with decreased mRNA levels as well as p27Kip1 promoter activity. 11,12-EET also stimulated the time-dependent phosphorylation of Akt and of the forkhead factors FOXO1 and FOXO3a, effects prevented by the phosphatidylinositol 3-kinase inhibitor LY 294002. Transfection of endothelial cells with either a dominant-negative or an "Akt-resistant"/constitutively active FOXO3a mutant reversed the 11,12-EET-induced down-regulation of p27Kip1, whereas transfection of a constitutive active Akt decreased p27Kip1 expression independently of the presence or absence of 11,12-EET. To determine whether these effects are involved in EET-induced proliferation, endothelial cells were transfected with the 11,12 EET-generating epoxygenase CYP2C9. Transfection of CYP2C9 elicited endothelial cell proliferation and this effect was inhibited in cells co-transfected with CYP2C9 and either a dominant-negative Akt or constitutively active FOXO3a. Reducing FOXO expression using RNA interference, on the other hand, attenuated p27Kip1 expression and stimulated endothelial cell proliferation. These results indicate that EET-induced endothelial cell proliferation is associated with the phosphatidylinositol 3-kinase/Akt-dependent phosphorylation and inactivation of FOXO factors and the subsequent decrease in expression of the cyclin-dependent kinase inhibitor p27Kip1. PMID- 12773536 TI - Crystal structure of the C-terminal 10-kDa subdomain of Hsc70. AB - The 70-kDa heat shock proteins (Hsp70), including the cognates (Hsc70), are molecular chaperones that prevent misfolding and aggregation of polypeptides in cells under both normal and stressed conditions. They are composed of two major structural domains: an N-terminal 44-kDa ATPase domain and a C-terminal 30-kDa substrate binding domain. The 30-kDa domain can be divided into an 18-kDa subdomain and a 10-kDa subdomain. Here we report the crystal structure of the 10 kDa subdomain of rat Hsc70 at 3.45 A. Its helical region adopted a helix-loop helix fold. This conformation is different from the equivalent subdomain of DnaK, the bacterial homologue of Hsc70. Moreover, in the crystalline state, the 10-kDa subdomain formed dimers. The results of gel filtration chromatography further supported the view that this subdomain was self-associated. Upon gel filtration, Hsc70 was found to exist as a mixture of monomers, dimers, and oligomers, but the 60-kDa fragment was predominantly found to exist as monomers. These findings suggest that the alpha-helical region of the 10-kDa subdomain dictates the chaperone self-association. PMID- 12773537 TI - Structure-function analysis of recombinant substrate protein 22 kDa (SP-22). A mitochondrial 2-CYS peroxiredoxin organized as a decameric toroid. AB - Bovine mitochondrial SP-22 is a member of the peroxiredoxin family of peroxidases. It belongs to the peroxiredoxin 2-Cys subgroup containing three cysteines at positions 47, 66, and 168. The cloning and overexpression in Escherichia coli of recombinant wild type SP-22 and its three cysteine mutants (C47S, C66S, and C168S) are reported. Purified His-tagged SP-22 was fully active with Cys-47 being confirmed as the catalytic residue. The enzyme forms a stable decameric toroid consisting of five basic dimeric units containing intermolecular disulfide bonds linking the catalytically active Cys-47 of one subunit and Cys 168 of the adjacent monomer. The disulfide bonds are not required for overall structural integrity. The toroidal units have average external and internal diameters of 15 and 7 nm, respectively, and can form stacks in a lateral arrangement of two or three rings. C47S had a pronounced tendency to stack in long tubular structures containing up to 60 rings. Further unusual structural features are the presence of radial spikes projecting from the external surface and ordered electron-dense material within the central cavity of the toroid. PMID- 12773539 TI - Role of proline residues in the folding of serine hydroxymethyltransferase. AB - Previous studies on the folding mechanism of Escherichia coli serine hydroxymethyltransferase (SHMT) showed that the final rate determining folding step was from an intermediate that contained two fully folded domains with N terminal segments of approximately 55 residues and interdomain segments of approximately 50 residues that were still solvent exposed and subject to proteolysis. The interdomain segment contains 3 Pro residues near its N terminus and 2 Pro residues near its C terminus. The 5 Pro residues were each mutated to both a Gly and Ala residue, and each mutant SHMT was purified and characterized with respect to kinetic properties, stability, secondary structure, and folding mechanism. The results showed that Pro214 and Pro218 near the N terminus of the interdomain segment are not critical for folding, stability, or activity. The P216A mutant also retained most of the characteristics of the native enzyme, but its folding rate was altered. However, the P216G mutant was severely compromised in folding into a catalytically competent enzyme. Mutation of both Pro258 and Pro264 had altered folding kinetics and resulted in enzymes that expressed little catalytic activity. The Phe257-Pro258 bond is cis in its configuration, and the P258A mutant SHMT showed reduced thermal stability. Pro216, Pro258, and Pro264 are conserved in all 53 known sequences of this enzyme. The results are discussed in terms of the role of each Pro residue in maintaining the structure and function of SHMT and a possible role in pyridoxal 5'-phosphate addition to the apo-enzyme. PMID- 12773538 TI - Evidence for distinct sodium-, dopamine-, and cocaine-dependent conformational changes in transmembrane segments 7 and 8 of the dopamine transporter. AB - Previously we obtained evidence based on engineering of Zn2+ binding sites that the extracellular parts of transmembrane segment 7 (TM7) and TM8 in the human dopamine transporter are important for transporter function. To further evaluate the role of this domain, we have employed the substituted cysteine accessibility method and performed 10 single cysteine substitutions at the extracellular ends of TM7 and TM8. The mutants were made in background mutants of the human dopamine transporter with either two (E2C) or five endogenous cysteines substituted (X5C) that render the transporter largely insensitive to cysteine modification. In two mutants (M371C and A399C), treatment with the sulfhydryl-reactive reagent [2 (trimethylammonium)-ethyl]methanethiosulfonate (MTSET) led to a substantial inhibition of [3H]dopamine uptake. In M371C this inactivation was enhanced by Na+ and blocked by dopamine. Inhibitors such as cocaine did not alter the effect of MTSET in M371C. The protection of M371C inactivation by dopamine required Na+. Because dopamine binding is believed to be Na+-independent, this suggests that dopamine induces a transport-associated conformational change that decreases the reactivity of M371C with MTSET. In contrast to M371C, cocaine decreased the reaction rate of A399C with MTSET, whereas dopamine had no effect. The protection by cocaine can either reflect that Ala-399 lines the cocaine binding crevice or that cocaine induces a conformational change that decreases the reactivity of A399C. The present findings add new functionality to the TM7/8 region by providing evidence for the occurrence of distinct Na+-, substrate-, and perhaps inhibitor-induced conformational changes critical for the proper function of the transporter. PMID- 12773540 TI - The Rho guanine nucleotide exchange factor Lsc homo-oligomerizes and is negatively regulated through domains in its carboxyl terminus that are absent in novel splenic isoforms. AB - Rho GTPases control fundamental cellular processes, including cytoskeletal reorganization and transcription. Rho guanine nucleotide exchange factors (GEFs) compose a large (>65) and diverse family of related proteins that activate Rho GTPases. Lsc/p115-RhoGEF is a Rho-specific GEF required for normal B and T lymphocyte function. Despite its essential role in lymphocytes, Lsc/p115-RhoGEF signaling in vivo is not well understood. To define Lsc/p115-RhoGEF signaling pathways in vivo, we set out to identify proteins that interact with regulatory regions of Lsc. The 146-amino acid C terminus of Lsc contains a predicted coiled coil domain, and we demonstrated that deletion of this C terminus confers a gain of function in vivo. Surprisingly, a yeast two-hybrid screen for proteins that interact with this regulatory C terminus isolated a larger C-terminal fragment of Lsc itself. Co-immunoprecipitation experiments in mammalian cells demonstrated that Lsc specifically homo-oligomerizes and that the coiled-coil domain in the C terminus is required for homo-oligomerization. Mutagenesis experiments revealed that homo-oligomerization and negative regulation are distinct functions of the C terminus. Two novel isoforms of Lsc found in the spleen lack portions of this C terminus, including the coiled-coil domain. Importantly, the C termini of both isoforms confer a gain of function and eliminate homo-oligomerization. These results define two important features of Lsc signaling. First, Lsc homo oligomerizes and is negatively regulated through domains in its C terminus; and second, functionally distinct isoforms of Lsc lacking these domains are present in the spleen. PMID- 12773541 TI - Plastocyanin is indispensable for photosynthetic electron flow in Arabidopsis thaliana. AB - Plastocyanin is a soluble copper-containing protein present in the thylakoid lumen, which transfers electrons to photosystem I. In the chloroplast of the flowering plant Arabidopsis thaliana, a cytochrome c6-like protein is present, which was recently suggested to function as an alternative electron carrier to plastocyanin. We show that Arabidopsis plants mutated in both of the two plastocyanin-coding genes and with a functional cytochrome c6 cannot grow photoautotrophically because of a complete block in light-driven electron transport. Even increased dosage of the gene encoding the cytochrome c6-like protein cannot complement the double mutant phenotype. This demonstrates that in Arabidopsis only plastocyanin can donate electrons to photosystem I in vivo. PMID- 12773542 TI - Nickel and extracellular acidification inhibit the water permeability of human aquaporin-3 in lung epithelial cells. AB - Nickel is a common cause of pneumoconiosis. Here, we show that nickel inactivates aquaporin (AQP)-3, the water channel expressed apically in epithelial cells of human terminal airways. Human AQP3 was transiently transfected into human lung cells, and water permeability was measured in transfected and neighboring untransfected cells. Incubation with NiCl2 rapidly, dose-dependently, and reversibly decreased water permeability in AQP3-expressing cells. Acidification of the extracellular medium also caused rapid, dose-dependent, and reversible inhibition of AQP3. Sensitivity of AQP3 to nickel was lower at alkaline pH than at neutral and acidic pH. Cells transfected with human AQP4 and AQP5, which are also expressed in airway epithelia, were insensitive to nickel and extracellular acidification. Zinc and cadmium, other common causes of pneumoconiosis, had no effect on the water permeability of AQP3. Three extracellular residues, Trp128, Ser152, and His241, were responsible for the blocking effect of nickel on human AQP3. Ser152 was identified as a common site for nickel and pH sensitivity. His53, Tyr124, and His154 were also involved in regulation of AQP3 by extracellular pH. In addition, the aromatic side chain of His154 was shown to be important for the water permeability of AQP3. Our results imply that nickel and extracellular pH may modulate lung water clearance and that defective water clearance may be an early component of nickel-induced lung disease. PMID- 12773543 TI - Leporipoxvirus Cu,Zn-superoxide dismutase (SOD) homologs are catalytically inert decoy proteins that bind copper chaperone for SOD. AB - Many Chordopoxviruses encode catalytically inactive homologs of cellular Cu-Zn superoxide dismutase (SOD). The biological function of these proteins is unknown, although the proteins encoded by Leporipoxviruses have been shown to promote a slow decline in the level of superoxide dismutase activity in virus-infected cells. To gain more insights into their function, we have further characterized the enzymatic and biochemical properties of a SOD homolog encoded by Shope fibroma virus. Shope fibroma virus SOD has retained the zinc binding properties of its cellular homolog, but cannot bind copper. Site-directed mutagenesis showed that it requires at least four amino acid substitutions to partially restore copper binding activity, but even these changes still did not restore catalytic activity. Reciprocal co-immunoprecipitation experiments showed that recombinant Shope fibroma virus SOD forms very stable complexes with cellular copper chaperones for SOD and these observations were confirmed using glutathione-S transferase tagged proteins. Similar viral SOD/chaperone complexes were formed in cells infected with a closely related myxoma virus, where we also noted that some of the SOD antigen co-localizes with mitochondrial markers using confocal fluorescence microscopy. About 2% of the viral SOD was subsequently detected in gradient-purified mitochondria extracted from virus-infected cells. These poxviral SOD homologs do not form stable complexes with cellular Cu,Zn-SOD or affect its concentration. We suggest that Leporipoxvirus SOD homologs are catalytically inert decoy proteins that are designed to interfere in the proper metallation and activation of cellular Cu,Zn-SOD. This reaction might be advantageous for tumorigenic poxviruses, since higher levels of superoxide have been proposed to have anti-apoptotic and tumorigenic activity. PMID- 12773544 TI - Production, characterization, and immunogenicity of a soluble rat single chain T cell receptor specific for an encephalitogenic peptide. AB - The encephalitogenic rat T cell clone C14 recognizes the myelin basic protein 69 89 peptide in the context of the RT1B major histocompatibility complex (MHC) class II molecule. Modeling of the C14 TCR molecule indicated that previously identified CDR3 motifs are likely to be central to interaction with MHC class II presented peptide. Here we report the cloning and expression of C14-derived single chain TCR (scTCR) molecules in an Escherichia coli expression system. The recombinant molecule consists of the Valpha2 domain connected to the Vbeta8.2 domain via a 15-residue linker. Soluble C14 scTCR was purified using conventional chromatography techniques and refolded by a rapid dilution procedure. C14 scTCR was able to bind soluble rat MHC class II molecules bearing covalently coupled Gp BP-(69-89) peptide, as analyzed using surface plasmon resonance. Immune recognition of the C14 scTCR protein as an antigen revealed that limited regions of the TCR may be more likely to induce responsiveness. PMID- 12773545 TI - Agrin is a chimeric proteoglycan with the attachment sites for heparan sulfate/chondroitin sulfate located in two multiple serine-glycine clusters. AB - Agrin is a large extracellular matrix protein that plays a key role in the formation and maintenance of the vertebrate neuromuscular junction. The amino acid sequence of agrin encodes a protein with a molecular size of 220 kDa, whereas SDS-PAGE shows a diffuse band around 400 kDa. Further studies showed that agrin is highly glycosylated and belongs to the family of heparan sulfate proteoglycans. By expressing different protein fragments, we localized the glycosaminoglycan (GAG) attachment sites to two locations within the agrin molecule. One site that is located between the seventh and eight follistatin-like domain includes 3 closely spaced serine-glycine (SG) consensus sequences and carries exclusively heparan sulfate side chains. The second site is located further downstream in the centrally located serine-threonine-rich domain and contains a cluster of 4 closely packed SG consensus sequences. This site predominantly carries chondroitin sulfate side chains. Investigating the contribution of individual serines in GAG priming by site-directed mutagenesis showed that each serine of the two SG clusters has the potential to carry GAGs. In accordance with the mixed GAG glycosylation of agrin peptide fragments, it was found that recombinant and in vivo-derived full-length agrin are not exclusively heparan sulfate proteoglycans but also carry chondroitin sulfate side chains. PMID- 12773546 TI - Fluorescent indicators for Akt/protein kinase B and dynamics of Akt activity visualized in living cells. AB - Akt/protein kinase B (PKB) is a serine/threonine kinase that regulates a variety of cellular responses. To provide information on the spatial and temporal dynamics of Akt/PKB activity, we have developed genetically encoded fluorescent indicators for Akt/PKB. The indicators contain two green fluorescent protein mutants, an Akt/PKB substrate domain, flexible linker sequence, and phosphorylation recognition domain. A phosphorylation of the substrate domain in the indicators caused change in the emission ratio based on fluorescent resonance energy transfer between the two green fluorescent protein mutants. To let the fluorescent indicators behave as endothelial nitric-oxide synthase and Bad, which are endogenous Akt/PKB substrates, they were fused with the Golgi target domain and mitochondria target domain, respectively. The indicators thus colocalized with the endogenous substrates conferred their susceptibilities to phosphorylation by Akt/PKB. We showed that the Golgi-localized indicator responded to the stimulation with 17beta-estradiol (E2) and insulin in endothelial cells. In addition, E2 elicited the phosphorylation of the mitochondria-localized indicator in the endothelial cells, but no phosphorylation was observed by E2 or by insulin of the diffusible indicator that has no targeting domain. The difference in the results with the three indicators suggests that the activated Akt/PKB is localized to subcellular compartments, including the Golgi apparatus and/or mitochondria, rather than diffusing in the cytosol, thereby efficiently phosphorylating its substrate proteins. E2 triggered the phosphorylation of the mitochondria-localized indicator, whereas insulin did not induce this phosphorylation, which suggests that the localization of the activated Akt/PKB to the mitochondria is directed differently between insulin and E2 via distinct mechanisms. PMID- 12773547 TI - Kinectin anchors the translation elongation factor-1 delta to the endoplasmic reticulum. AB - Kinectin has been proposed to be a membrane anchor for kinesin on intracellular organelles. A kinectin isoform that lacks a major portion of the kinesin-binding domain does not bind kinesin but interacts with another resident of the endoplasmic reticulum, the translation elongation factor-1 delta (EF-1 delta). This was shown by yeast two-hybrid analysis and a number of in vitro and in vivo assays. EF-1 delta provides the guanine nucleotide exchange activities on EF-1 alpha during elongation step of protein synthesis. The minimal EF-1 delta-binding domain on kinectin resides within a conserved region present in all the kinectin isoforms. Overexpression of the kinectin fragments in vivo disrupted the intracellular localization of EF-1 delta proteins. This report provides evidence of an alternative kinectin function as the membrane anchor for EF-1 delta on the endoplasmic reticulum and provides clues to the EF-1 complex assembly and anchorage on the endoplasmic reticulum. PMID- 12773548 TI - Reactivation of formyl peptide receptors triggers the neutrophil NADPH-oxidase but not a transient rise in intracellular calcium. AB - In neutrophils, coupling of chemoattractants to their cell surface receptor at low temperature (W). Treatment with depsipeptide, an inhibitor of histone deacetylase, was unable to induce p53-independent p21(Cip1) expression because the promoter of p21(Cip1) in these cells is hypermethylated. By analyzing luciferase activity of transfected p21(Cip1) promoter vectors, we demonstrate that depsipeptide functions on Sp1 binding sites to induce p21(Cip1) expression. We hypothesize that hypermethylation may interfere with Sp1/Sp3 binding. By using an electrophoretic mobility shift assay, we show that, although methylation within the consensus Sp1 binding site did not reduce Sp1/Sp3 binding, methylation outside of the consensus Sp1 element induced a significant decrease in Sp1/Sp3 binding. Depsipeptide induced p21(Cip1) expression was reconstituted when cells were pretreated with 5 aza-2'-deoxycytidine. Our data suggest, for the first time, that hypermethylation around the consensus Sp1-binding sites may directly reduce Sp1/Sp3 binding, therefore leading to a reduced p21(Cip1) expression in response to depsipeptide treatment. PMID- 12773552 TI - Synergism between calcium and cyclic GMP in cyclic AMP response element-dependent transcriptional regulation requires cooperation between CREB and C/EBP-beta. AB - Calcium induces transcriptional activation of the fos promoter by activation of the cyclic AMP response element (CRE)-binding protein (CREB), and in some cells its effect is enhanced synergistically by cyclic GMP (cGMP) through an unknown mechanism. We observed calcium-cGMP synergism in neuronal and osteogenic cells which express type II cGMP-dependent protein kinase (G-kinase); the effect on the fos promoter was mediated by the CRE and proportional to G-kinase activity. Dominant negative transcription factors showed involvement of CREB- and C/EBP related proteins but not of AP-1. Expression of C/EBP-beta but not C/EBP-alpha or -delta enhanced the effects of calcium and cGMP on a CRE-dependent reporter gene. The transactivation potential of full-length CREB fused to the DNA-binding domain of Gal4 was increased synergistically by calcium and cGMP, and overexpression of C/EBP-beta enhanced the effect, while a dominant negative C/EBP inhibited it. With a mammalian two-hybrid system, coimmunoprecipitation experiments, and in vitro binding studies, we demonstrated that C/EBP-beta and CREB interacted directly; this interaction involved the C terminus of C/EBP-beta but occurred independently of CREB's leucine zipper domain. CREB Ser(133) phosphorylation was stimulated by calcium but not by cGMP; in cGMP-treated cells, (32)PO(4) incorporation into C/EBP-beta was decreased and C/EBP-beta/CRE complexes were increased, suggesting regulation of C/EBP-beta functions by G-kinase-dependent dephosphorylation. C/EBP-beta and CREB associated with the fos promoter in intact cells, and the amount of promoter-associated C/EBP-beta was increased by calcium and cGMP. We conclude that calcium and cGMP transcriptional synergism requires cooperation of CREB and C/EBP-beta, with calcium and cGMP modulating the phosphorylation states of CREB and C/EBP-beta, respectively. PMID- 12773553 TI - The RNA binding domain of Jerky consists of tandemly arranged helix-turn helix/homeodomain-like motifs and binds specific sets of mRNAs. AB - A deficit in the Jerky protein in mice causes recurrent seizures reminiscent of temporal lobe epilepsy. Jerky is present in mRNA particles in neurons. We show that the N-terminal 168 amino acids of Jerky are necessary and sufficient for mRNA binding. The binding domain is similar to the two tandemly arranged homeodomain-like helix-turn-helix DNA binding motifs of centromere binding protein B. The putative helix-turn-helix motifs of Jerky can also bind double stranded DNA and represent a novel mammalian RNA/DNA binding domain. Microarray analysis identified mRNAs encoding proteins involved in ribosome assembly and cellular stress response that specifically bound to the RNA binding domain of Jerky both in vitro and in vivo. These data suggest that epileptogenesis in Jerky deficient mice most likely involves pathways associated with ribosome biogenesis and neuronal survival and/or apoptosis. PMID- 12773554 TI - Yeast Lsm1p-7p/Pat1p deadenylation-dependent mRNA-decapping factors are required for brome mosaic virus genomic RNA translation. AB - Previously, we used the ability of the higher eukaryotic positive-strand RNA virus brome mosaic virus (BMV) to replicate in yeast to show that the yeast LSM1 gene is required for recruiting BMV RNA from translation to replication. Here we extend this observation to show that Lsm1p and other components of the Lsm1p Lsm7p/Pat1p deadenylation-dependent mRNA decapping complex were also required for translating BMV RNAs. Inhibition of BMV RNA translation was selective, with no effect on general cellular translation. We show that viral genomic RNAs suitable for RNA replication were already distinguished from nonreplication templates at translation, well before RNA recruitment to replication. Among mRNA turnover pathways, only factors specific for deadenylated mRNA decapping were required for BMV RNA translation. Dependence on these factors was not only a consequence of the nonpolyadenylated nature of BMV RNAs but also involved the combined effects of the viral 5' and 3' noncoding regions and 2a polymerase open reading frame. High-resolution sucrose density gradient analysis showed that, while mutating factors in the Lsm1p-7p/Pat1p complex completely inhibited viral RNA translation, the levels of viral RNA associated with ribosomes were only slightly reduced in mutant yeast. This polysome association was further verified by using a conditional allele of essential translation initiation factor PRT1, which markedly decreased polysome association of viral genomic RNA in the presence or absence of an LSM7 mutation. Together, these results show that a defective Lsm1p 7p/Pat1p complex inhibits BMV RNA translation primarily by stalling or slowing the elongation of ribosomes along the viral open reading frame. Thus, factors in the Lsm1p-7p/Pat1p complex function not only in mRNA decapping but also in translation, and both translation and recruitment of BMV RNAs to viral RNA replication are regulated by a cell pathway that transfers mRNAs from translation to degradation. PMID- 12773555 TI - TATA-binding protein-like protein (TLP/TRF2/TLF) negatively regulates cell cycle progression and is required for the stress-mediated G(2) checkpoint. AB - The TATA-binding protein (TBP) is a universal transcription factor required for all of the eukaryotic RNA polymerases. In addition to TBP, metazoans commonly express a distantly TBP-related protein referred to as TBP-like protein (TLP/TRF2/TLF). Although the function of TLP in transcriptional regulation is not clear, it is known that TLP is required for embryogenesis and spermiogenesis. In the present study, we investigated the cellular functions of TLP by using TLP knockout chicken DT40 cells. TLP was found to be dispensable for cell growth. Unexpectedly, TLP-null cells exhibited a 20% elevated cell cycle progression rate that was attributed to shortening of the G(2) phase. This indicates that TLP functions as a negative regulator of cell growth. Moreover, we found that TLP mainly existed in the cytoplasm and was translocated to the nucleus restrictedly at the G(2) phase. Ectopic expression of nuclear localization signal-carrying TLP resulted in an increase (1.5-fold) in the proportion of cells remaining in the G(2)/M phase and apoptotic state. Notably, TLP-null cells showed an insufficient G(2) checkpoint when the cells were exposed to stresses such as UV light and methyl methanesulfonate, and the population of apoptotic cells after stresses decreased to 40%. These phenomena in G(2) checkpoint regulation are suggested to be p53 independent because p53 does not function in DT40 cells. Moreover, TLP was transiently translocated to the nucleus shortly (15 min) after stress treatment. The expression of several stress response and cell cycle regulatory genes drifted in a both TLP- and stress-dependent manner. Nucleus-translocating TLP is therefore thought to work by checking cell integrity through its transcription regulatory ability. TLP is considered to be a signal-transducing transcription factor in cell cycle regulation and stress response. PMID- 12773557 TI - Xkid is degraded in a D-box, KEN-box, and A-box-independent pathway. AB - During mitosis, the Xenopus chromokinesin Kid (Xkid) provides the polar ejection forces needed at metaphase for chromosome congression, and its degradation is required at anaphase to induce chromosome segregation. Despite the fact that the degradation of Xkid at anaphase seems to be a key regulatory factor to induce chromosome movement to the poles, little is known about the mechanisms controlling this proteolysis. We investigated here the degradation pathway of Xkid. We demonstrate that Xkid is degraded both in vitro and in vivo by APC/Cdc20 and APC/Cdh1. We show that, despite the presence of five putative D-box motifs in its sequence, Xkid is proteolyzed in a D-box-independent manner. We identify a domain within the C terminus of this chromokinesin, with sequence GxEN, whose mutation completely stabilizes this protein by both APC/Cdc20 and APC/Cdh1. Moreover, we show that this degradation sequence acts as a transposable motif and induces the proteolysis of a GST-GXEN fusion protein. Finally, we demonstrate that both a D-box and a GXEN-containing peptides completely block APC-dependent degradation of cyclin B and Xkid, indicating that the GXEN domain might mediate the recognition and association of Xkid with the APC. PMID- 12773556 TI - Effect of damage type on stimulation of human excision nuclease by SWI/SNF chromatin remodeling factor. AB - To investigate the repair of different types of DNA lesions in chromatin, we prepared mononucleosomes containing an acetylaminofluorene-guanine adduct (AAF G), a (6-4) photoproduct, or a cyclobutane pyrimidine dimer (CPD) and measured the repair of these lesions by reconstituted 6-factor human excision nuclease. We find that incorporation into nucleosomes inhibits the repair of CPD more severely than repair of the AAF-G adduct and the (6-4) photoproduct. Equally important, we find that SWI/SNF stimulates the removal of AAF-G and (6-4) photoproduct but not of CPD from nucleosomal DNA. These results shed new light on the low rate of repair of CPDs in human cells in vivo. PMID- 12773558 TI - Regulation and substrate specificity of the SR protein kinase Clk/Sty. AB - SR proteins constitute a family of splicing factors that play key roles in both constitutive and regulated splicing in metazoan organisms. The proteins are extensively phosphorylated, and kinases capable of phosphorylating them have been identified. However, little is known about how these kinases function, for example, whether they target specific SR proteins or whether the kinases themselves are regulated. Here we describe properties of one such kinase, Clk/Sty, the founding member of the Clk/Sty family of dual-specificity kinases. Clk/Sty is autophosphorylated on both Ser/Thr and Thr residues, and using both direct kinase assays and SR protein-dependent splicing assays, we have analyzed the effects of each type of modification. We find not only that the pattern of phosphorylation on a specific SR protein substrate, ASF/SF2, is modulated by autophosphorylation but also that the ability of Clk/Sty to recognize different SR proteins is influenced by the extent and nature of autophosphorylation. Strikingly, phosphorylation of ASF/SF2 is sensitive to changes in Tyr, but not Ser/Thr, autophosphorylation while that of SC35 displays the opposite pattern. In contrast, phosphorylation of a third SR protein, SRp40, is unaffected by autophosphorylation. We also present biochemical data indicating that as expected for a factor directly involved in splicing control (but in contrast to recent reports), Clk/Sty is found in the nucleus of several different cell types. PMID- 12773559 TI - Role of the promoter in maintaining transcriptionally active chromatin structure and DNA methylation patterns in vivo. AB - Establishment and maintenance of differential chromatin structure between transcriptionally competent and repressed genes are critical aspects of transcriptional regulation. The elements and mechanisms that mediate formation and maintenance of these chromatin states in vivo are not well understood. To examine the role of the promoter in maintaining chromatin structure and DNA methylation patterns of the transcriptionally active X-linked HPRT locus, 323 bp of the endogenous human HPRT promoter (from position -222 to +102 relative to the translation start site) was replaced by plasmid sequences by homologous recombination in cultured HT-1080 male fibrosarcoma cells. The targeted cells, which showed no detectable HPRT transcription, were then assayed for effects on DNase I hypersensitivity, general DNase I sensitivity, and DNA methylation patterns across the HPRT locus. In cells carrying the deletion, significantly diminished DNase I hypersensitivity in the 5' flanking region was observed compared to that in parental HT-1080 cells. However, general DNase I sensitivity and DNA methylation patterns were found to be very similar in the mutated cells and in the parental cells. These findings suggest that the promoter and active transcription play a relatively limited role in maintaining transcriptionally potentiated epigenetic states. PMID- 12773561 TI - Spatial organization of protein-RNA interactions in the branch site-3' splice site region during pre-mRNA splicing in yeast. AB - A series of efficiently spliced pre-mRNA substrates containing single 4 thiouridine residues were used to monitor RNA-protein interactions involving the branch site-3' splice site-3' exon region during yeast pre-mRNA splicing through cross-linking analysis. Prior to the assembly of the prespliceosome, Mud2p and the branch point bridging protein cross-link to a portion of this region in an ATP-independent fashion. Assembly of the prespliceosome leads to extensive cross linking of the U2-associated protein Hsh155p to this region. Following the first step of splicing and in a manner independent of Prp16p, the U5 small nuclear ribonucleoprotein particle-associated protein Prp8p also associates extensively with the branch site-3' splice site-3' exon region. The subsequent cross-linking of Prp16p to the lariat intermediate is restricted to the 3' splice site and the adjacent 3' exon sequence. Using modified substrates to either mutationally or chemically block the second step, we found that the association of Prp22p with the lariat intermediate represents an authentic transient intermediate and appears to be restricted to the last eight intron nucleotides. Completion of the second step leads to the cross-linking of an unidentified approximately 80-kDa protein near the branch site sequence, suggesting a potential role for this protein in a later step in intron metabolism. Taken together, these data provide a detailed portrayal of the dynamic associations of proteins with the branch site 3' splice site region during spliceosome assembly and catalysis. PMID- 12773560 TI - Deacetylase activity is required for recruitment of the basal transcription machinery and transactivation by STAT5. AB - The signal transducer and activator of transcription STAT5 plays a major role in the cellular response to cytokines, but the mechanism by which it activates transcription remains poorly understood. We show here that deacetylase inhibitors (trichostatin A, suberoylanilide hydroxamic acid, and sodium butyrate) prevent induction of endogenous STAT5 target genes, implying that a deacetylase activity is required for that process. Microarray analyses revealed that this requirement is common to all STAT5 target genes. Using chromatin immunoprecipitation, we show that, following STAT5 DNA binding, deacetylase inhibitors block transcription initiation by preventing recruitment of the basal transcription machinery. This inhibition is not due to effects on histone H3 and H4 acetylation or chromatin remodeling within the promoter region. This novel mechanism of transactivation by STAT5 provides a rationale for the use of deacetylase inhibitors for therapeutic intervention in STAT5-associated cancers. PMID- 12773562 TI - Regulation of subnuclear localization is associated with a mechanism for nuclear receptor corepression by RIP140. AB - Regulation of gene transcription by nuclear receptors involves association with numerous coregulators. Receptor-interacting protein 140 (RIP140) is a corepressor that negatively regulates the ligand-induced activity of several nuclear receptors, including the glucocorticoid receptor (GR). In the present study, we have characterized the role of the intranuclear localization of RIP140 in its corepressor activity. In the absence of ligand-activated GR, RIP140 is localized in small nuclear foci targeted by a 40-amino-acid-long sequence. Although the focus-targeting domain overlaps with a binding sequence for the corepressor CtBP (C-terminal binding protein), interaction with CtBP is not involved in the localization. RIP140 foci do not correspond to PML bodies but partly colocalize with domains harboring the corepressor SMRT. Upon ligand binding, GR and RIP140 are redistributed to large nuclear domains distinct from the RIP140 foci. The redistribution requires regions of RIP140 with corepressor activity, as well as the DNA-binding domain of GR. Furthermore, we show that full RIP140 corepressor activity is contributed both by C-terminal receptor-binding LXXLL motifs and interaction with the CtBP corepressor. In conclusion, our results suggest that the corepressor function of RIP140 is multifaceted and involves binding to nuclear receptors, as well as additional functions mediated by the formation and intranuclear relocalization of a repressive protein complex. PMID- 12773563 TI - CD33/Siglec-3 binding specificity, expression pattern, and consequences of gene deletion in mice. AB - Mouse CD33/Siglec-3 (mCD33) is the apparent ortholog of human CD33/Siglec-3 (hCD33), a member of the Siglec (sialic acid-binding Ig superfamily lectin) family of sialic acid-recognizing cell-surface lectins. We examined the binding specificity and expression pattern of mCD33 and explored its functions by generating mice deficient in this molecule. Like hCD33, mCD33 is expressed on myeloid precursors in the bone marrow, albeit mostly in the more mature stages of the granulocytic lineage. Moreover, unlike hCD33, mCD33 in peripheral blood is primarily expressed on granulocytes. Also, unlike hCD33, mCD33 did not bind to alpha2-3- or alpha2-6-linked sialic acids on lactosamine units. Instead, it showed distinctive sialic acid-dependent binding only to the short O-linked glycans of certain mucins and weak binding to the sialyl-Tn epitope. Binding was enhanced by removal of 9-O-acetyl groups and attenuated by truncation of the glycerol-like side chain of sialic acids. Mice deficient in CD33 were viable and fertile in a controlled-access specific-pathogen-free vivarium, showed no major morphological or histological abnormalities, had no changes in bone marrow or peripheral leukocyte subpopulations, and had very minor differences in biochemical and erythrocyte parameters. Cellular responses to intraperitoneally injected proinflammatory stimulants, as well as subsequent interleukin-6 secretion, were also apparently unaffected. These results indicate substantial species differences in CD33 expression patterns and ligand recognition and suggest functional degeneracy between mCD33 and the other CD33-related Siglec proteins expressed on cells of the myeloid lineage. PMID- 12773564 TI - Methylation of histone H3 by Set2 in Saccharomyces cerevisiae is linked to transcriptional elongation by RNA polymerase II. AB - Set2 methylates Lys36 of histone H3. We show here that yeast Set2 copurifies with RNA polymerase II (RNAPII). Chromatin immunoprecipitation analyses demonstrated that Set2 and histone H3 Lys36 methylation are associated with the coding regions of several genes that were tested and correlate with active transcription. Both depend, as well, on the Paf1 elongation factor complex. The C terminus of Set2, which contains a WW domain, is also required for effective Lys36 methylation. Deletion of CTK1, encoding an RNAPII CTD kinase, prevents Lys36 methylation and Set2 recruitment, suggesting that methylation may be triggered by contact of the WW domain or C terminus of Set2 with Ser2-phosphorylated CTD. A set2 deletion results in slight sensitivity to 6-azauracil and much less beta-galactosidase produced by a reporter plasmid, resulting from a defect in transcription. In synthetic genetic array (SGA) analysis, synthetic growth defects were obtained when a set2 deletion was combined with deletions of all five components of the Paf1 complex, the chromodomain elongation factor Chd1, the putative elongation factor Soh1, the Bre1 or Lge1 components of the histone H2B ubiquitination complex, or the histone H2A variant Htz1. SET2 also interacts genetically with components of the Set1 and Set3 complexes, suggesting that Set1, Set2, and Set3 similarly affect transcription by RNAPII. PMID- 12773565 TI - RhoE binds to ROCK I and inhibits downstream signaling. AB - RhoE belongs to the Rho GTPase family, the members of which control actin cytoskeletal dynamics. RhoE induces stress fiber disassembly in a variety of cell types, whereas RhoA stimulates stress fiber assembly. The similarity of RhoE and RhoA sequences suggested that RhoE might compete with RhoA for interaction with its targets. Here, we show that RhoE binds ROCK I but none of the other RhoA targets tested. The interaction of RhoE with ROCK I was confirmed by coimmunoprecipitation of the endogenous proteins, and the two proteins colocalized on the trans-Golgi network in COS-7 cells. Although RhoE and RhoA were not able to bind ROCK I simultaneously, RhoE bound to the amino-terminal region of ROCK I encompassing the kinase domain, at a site distant from the carboxy-terminal RhoA-binding site. Overexpression of RhoE inhibited ROCK I induced stress fiber formation and phosphorylation of the ROCK I target myosin light chain phosphatase. These data suggest that RhoE induces stress fiber disassembly by directly binding ROCK I and inhibiting it from phosphorylating downstream targets. PMID- 12773566 TI - Molecular basis of p53 functional inactivation by the leukemic protein MLL-ELL. AB - The Eleven Lysine-rich Leukemia (ELL) gene undergoes translocation and fuses in frame to the Multiple Lineage Leukemia (MLL) gene in a substantial proportion of patients suffering from acute forms of leukemia. Molecular mechanisms of cellular transformation by the MLL-ELL fusion are not well understood. Although both MLL ELL and wild-type ELL can reduce functional activity of p53 tumor suppressor, our data reveal that MLL-ELL is a much more efficient inhibitor of p53 than is wild type ELL. We also demonstrate for the first time that ELL extreme C terminus [ELL(eCT)] is required for the recruitment of p53 into MLL-ELL nuclear foci and is both necessary and sufficient for the MLL-ELL inhibition of p53-mediated induction of p21 and apoptosis. Finally, our results demonstrate that MLL-ELL requires the presence of intact ELL(eCT) in order to disrupt p53 interactions with p300/CBP coactivator and thus significantly reduce p53 acetylation in vivo. Since ELL(eCT) has recently been shown to be both necessary and sufficient for MLL-ELL-mediated transformation of normal blood progenitors, our data correlate ELL(eCT) contribution to MLL-ELL transformative effects with its ability to functionally inhibit p53. PMID- 12773567 TI - PML colocalizes with and stabilizes the DNA damage response protein TopBP1. AB - The PML tumor suppressor gene is consistently disrupted by t(15;17) in patients with acute promyelocytic leukemia. Promyelocytic leukemia protein (PML) is a multifunctional protein that plays essential roles in cell growth regulation, apoptosis, transcriptional regulation, and genome stability. Our study here shows that PML colocalizes and associates in vivo with the DNA damage response protein TopBP1 in response to ionizing radiation (IR). Both PML and TopBP1 colocalized with the IR-induced bromodeoxyuridine single-stranded DNA foci. PML and TopBP1 also colocalized with Rad50, Brca1, ATM, Rad9, and BLM. IR and interferon (IFN) coinduce the expression levels of both TopBP1 and PML. In PML-deficient NB4 cells, TopBP1 was unable to form IR-induced foci. All-trans-retinoic acid induced reorganization of the PML nuclear body (NB) and reappearance of the IR-induced TopBP1 foci. Inhibition of PML expression by siRNA is associated with a significant decreased in TopBP1 expression. Furthermore, PML-deficient cells express a low level of TopBP1, and its expression cannot be induced by IR or IFN. Adenovirus-mediated overexpression of PML in PML(-/-) mouse embryo fibroblasts substantially increased TopBP1 expression, which colocalized with the PML NBs. These studies demonstrated a mechanism of PML-dependent expression of TopBP1. PML overexpression induced TopBP1 protein but not the mRNA expression. Pulse-chase labeling analysis demonstrated that PML overexpression stabilized the TopBP1 protein, suggesting that PML plays a role in regulating the stability of TopBP1 in response to IR. Together, our findings demonstrate that PML regulates TopBP1 functions by association and stabilization of the protein in response to IR induced DNA damage. PMID- 12773568 TI - Cysteine 64 of Ref-1 is not essential for redox regulation of AP-1 DNA binding. AB - Ref-1 participates in DNA repair as well as in redox regulation of transcription factor function. The redox function of Ref-1 involves reduction of oxidized cysteine residues within the DNA binding domains of several transcription factors, including Fos and Jun. Reduction of these residues is required for DNA binding, providing a redox-dependent mechanism for regulation of target gene expression. Previous in vitro studies implicated cysteine 65 of human Ref-1 (cysteine 64 of mouse Ref-1) as the redox catalytic site. We analyzed the in vivo role of cysteine 64 in redox regulation of AP-1 activity by introducing a cysteine-to-alanine point mutation into the endogenous mouse Ref-1 gene (ref 1(C64A)). Unlike Ref-1 null mice, which die very early in embryonic development, homozygous ref-1(C64A) mice are viable, they survive to normal life expectancy, and they display no overt abnormal phenotype. Although Ref-1 provides the major AP-1-reducing activity in murine cells, ref-1(C64A) cells retain normal levels of endogenous AP-1 DNA binding activity in vivo as well as normal Fos- and Jun reducing activity in vitro. These results demonstrate that Ref-1 cysteine 64/65 is not required for redox regulation of AP-1 DNA binding in vivo, and they challenge previous hypotheses regarding the mechanism by which Ref-1 regulates the redox-dependent activity of specific transcription factors. PMID- 12773569 TI - JAM4, a junctional cell adhesion molecule interacting with a tight junction protein, MAGI-1. AB - MAGI-1 is a membrane-associated guanylate kinase protein at tight junctions in epithelial cells. It interacts with various molecules and functions as a scaffold protein at cell junctions. We report here a novel MAGI-1-binding protein that we named junctional adhesion molecule 4 (JAM4). JAM4 belongs to an immunoglobulin protein family. JAM4 was colocalized with ZO-1 in kidney glomeruli and in intestinal epithelial cells. Biochemical in vitro studies revealed that JAM4 bound to MAGI-1 but not to ZO-1, whereas JAM1 did not bind to MAGI-1. JAM4 and MAGI-1 interacted with each other and formed clusters in COS-7 cells when coexpressed. JAM4 mediated calcium-independent homophilic adhesion and was accumulated at cell-cell contacts when expressed in L cells. MAGI-1, ZO-1, and occludin were recruited to JAM4-based cell contacts. JAM4 also reduced the permeability of CHO cell monolayers. MAGI-1 strengthened JAM4-mediated cell adhesion in L cells and sealing effects in CHO cells. These findings suggest that JAM4 together with MAGI-1 provides an adhesion machinery at tight junctions, which may regulate the permeability of kidney glomerulus and small intestinal epithelial cells. PMID- 12773571 TI - Targeted disruption of the murine Bin1/Amphiphysin II gene does not disable endocytosis but results in embryonic cardiomyopathy with aberrant myofibril formation. AB - The mammalian Bin1/Amphiphysin II gene encodes an assortment of alternatively spliced adapter proteins that exhibit markedly divergent expression and subcellular localization profiles. Bin1 proteins have been implicated in a variety of different cellular processes, including endocytosis, actin cytoskeletal organization, transcription, and stress responses. To gain insight into the physiological functions of the Bin1 gene, we have disrupted it by homologous recombination in the mouse. Bin1 loss had no discernible impact on either endocytosis or phagocytosis in mouse embryo-derived fibroblasts and macrophages, respectively. Similarly, actin cytoskeletal organization, proliferation, and apoptosis in embryo fibroblasts were all unaffected by Bin1 loss. In vivo, however, Bin1 loss resulted in perinatal lethality. Bin1 has been reported to affect muscle cell differentiation and T-tubule formation. No striking histological abnormalities were evident in skeletal muscle of Bin1 null embryos, but severe ventricular cardiomyopathy was observed in these embryos. Ultrastructurally, myofibrils in ventricular cardiomyocytes of Bin1 null embryos were severely disorganized. These results define a developmentally critical role for the Bin1 gene in cardiac muscle development. PMID- 12773570 TI - Effects of rho kinase and actin stress fibers on sustained extracellular signal regulated kinase activity and activation of G(1) phase cyclin-dependent kinases. AB - We recently reported that Rho kinase is required for sustained ERK signaling and the consequent mid-G(1) phase induction of cyclin D1 in fibroblasts. The results presented here indicate that these Rho kinase effects are mediated by the formation of stress fibers and the consequent clustering of alpha5beta1 integrin. Mechanistically, alpha5beta1 signaling and stress fiber formation allowed for the sustained activation of MEK, and this effect was mediated upstream of Ras-GTP loading. Interestingly, disruption of stress fibers with ML-7 led to G(1) phase arrest while comparable disruption of stress fibers with Y27632 (an inhibitor of Rho kinase) or dominant-negative Rho kinase led to a more rapid progression through G(1) phase. Inhibition of either MLCK or Rho kinase blocked sustained ERK signaling, but only Rho kinase inhibition allowed for the induction of cyclin D1 and activation of cdk4 via Rac/Cdc42. The levels of cyclin E, cdk2, and their major inhibitors, p21(cip1) and p27(kip1), were not affected by inhibition of MLCK or Rho kinase. Overall, our results indicate that Rho kinase-dependent stress fiber formation is required for sustained activation of the MEK/ERK pathway and the mid-G(1) phase induction of cyclin D1, but not for other aspects of cdk4 or cdk2 activation. They also emphasize that G(1) phase cell cycle progression in fibroblasts does not require stress fibers if Rac/Cdc42 signaling is allowed to induce cyclin D1. PMID- 12773572 TI - TAF10 (TAF(II)30) is necessary for TFIID stability and early embryogenesis in mice. AB - TAF10 (formerly TAF(II)30), is a component of TFIID and the TATA box-binding protein (TBP)-free TAF-containing complexes (TFTC/PCAF/STAGA). To investigate the physiological function of TAF10, we disrupted its gene in mice by using a Cre recombinase/LoxP strategy. Interestingly, no TAF10(-/-) animals were born from intercrosses of TAF10(+/-) mice, indicating that TAF10 is required for embryogenesis. TAF10(-/-) embryos developed to the blastocyst stage, implanted, but died shortly after ca. 5.5 days postcoitus. Surprisingly, trophoblast cells from TAF10(-/-) blastocysts were viable, whereas inner cell mass cells failed to survive, highlighting that TAF10 is not generally required for transcription in all cells. TAF10-deficient cells express normal levels of TBP and TAFs other than TAF10 but contain only partially formed TFIID, are endocycle arrested, and have undetectable levels of transcription. Thus, our results demonstrate that TAF10 is required for TFIID stability, cell cycle progression, and transcription in the early mouse embryo. PMID- 12773573 TI - Molecular origins for the dominant negative function of human glucocorticoid receptor beta. AB - This study molecularly elucidates the basis for the dominant negative mechanism of the glucocorticoid receptor (GR) isoform hGRbeta, whose overexpression is associated with human glucocorticoid resistance. Using a series of truncated hGRalpha mutants and sequential mutagenesis to generate a series of hGRalpha/beta hybrids, we find that the absence of helix 12 is neither necessary nor sufficient for the GR dominant negative phenotype. Moreover, we have localized the dominant negative activity of hGRbeta to two residues and found that nuclear localization, in addition to heterodimerization, is a critical feature of the dominant negative activity. Molecular modeling of wild-type and mutant hGRalpha and hGRbeta provides structural insight and a potential physical explanation for the lack of hormone binding and the dominant negative actions of hGRbeta. PMID- 12773574 TI - Altered skeletal muscle phenotypes in calcineurin Aalpha and Abeta gene-targeted mice. AB - Calcineurin is a calcium-regulated serine-threonine protein phosphatase that controls developmental and inducible biological responses in diverse cell types, in part through activation of the transcription factor nuclear factor of activated T cells (NFAT). In skeletal muscle, calcineurin has been implicated in the regulation of myoblast differentiation, hypertrophy of mature myofibers, and fiber type switching in response to alterations in intracellular calcium concentration. However, considerable disagreement persists about the functional role of calcineurin signaling in each of these processes. Here we evaluated the molecular phenotypes of skeletal muscle from both calcineurin Aalpha and calcineurin Abeta gene-targeted mice. Calcineurin Aalpha was observed to be the predominant catalytic isoform expressed in nearly all skeletal muscles examined. Neither calcineurin Aalpha or Abeta null mice showed any gross growth-related alterations in skeletal muscle, nor was fiber size or number altered in glycolytic/fast muscle types. In contrast, both calcineurin Aalpha and Abeta gene targeted mice demonstrated an alteration in myofiber number in the soleus, an oxidative/slow-type muscle. More significantly, calcineurin Aalpha and Abeta gene targeted mice showed a dramatic down-regulation in the oxidative/slow fiber type program in multiple muscles (both slow and fast). Associated with this observation, NFAT-luciferase reporter transgenic mice showed significantly greater activity in slow fiber-containing muscles than in fast. However, only calcineurin Aalpha null mice showed a defect in NFAT nuclear occupancy or NFAT luciferase transgene activity in vivo. Collectively, our results suggest that calcineurin signaling plays a critical role in regulating skeletal muscle fiber type switching but not hypertrophy. Our results also suggest that fiber type switching occurs through an NFAT-independent mechanism. PMID- 12773576 TI - A novel jmjC domain protein modulates heterochromatization in fission yeast. AB - The heterochromatin domain at the mat locus of Schizosaccharomyces pombe is bounded by the IR-L and IR-R barriers. A genetic screen for mutations that promote silencing beyond IR-L revealed a novel gene named epe1, encoding a conserved nuclear protein with a jmjC domain. Disruption of epe1 promotes continuous spreading of heterochromatin-associated histone modifications and Swi6 binding to chromatin across heterochromatic barriers. It also enhances position effect variegation at heterochromatic domains, suppresses mutations in silencing genes, and stabilizes the repressed epigenetic state at the mat locus. However, it does not enhance silencing establishment. Our analysis suggests that the jmjC domain is essential for Epe1 activity and that Epe1 counteracts transcriptional silencing by negatively affecting heterochromatin stability. Consistent with this proposition, the meiotic stability of established heterochromatin beyond IR-L is diminished by Epe1 activity, and overexpression of Epe1 disrupts heterochromatin through acetylation of H3-K9 and H3-K14 and methylation of H3-K4. Furthermore, overexpression of Epe1 elevates the rate of chromosome loss. We propose that Epe1 helps control chromatin organization by down-regulating the stability of epigenetic marks that govern heterochromatization. PMID- 12773575 TI - The putative GTPases Nog1p and Lsg1p are required for 60S ribosomal subunit biogenesis and are localized to the nucleus and cytoplasm, respectively. AB - We characterized two essential putative GTPases, Nog1p and Lsg1p, that are found associated with free 60S ribosomal subunits affinity purified with the nuclear export adapter Nmd3p. Nog1p and Lsg1p are nucleolar and cytoplasmic, respectively, and are not simultaneously on the same particle, reflecting the path of Nmd3p shuttling in and out of the nucleus. Conditional mutants of both NOG1 and LSG1 are defective in 60S subunit biogenesis and display diminished levels of 60S subunits at restrictive temperature. Mutants of both genes also accumulate the 60S ribosomal reporter Rpl25-eGFP in the nucleolus, suggesting that both proteins are needed for subunit export from the nucleolus. Since Lsg1p is cytoplasmic, its role in nuclear export is likely to be indirect. We suggest that Lsg1p is needed to recycle an export factor(s) that shuttles from the nucleus associated with the nascent 60S subunit. PMID- 12773577 TI - Heart and liver defects and reduced transforming growth factor beta2 sensitivity in transforming growth factor beta type III receptor-deficient embryos. AB - The type III transforming growth factor beta (TGFbeta) receptor (TbetaRIII) binds both TGFbeta and inhibin with high affinity and modulates the association of these ligands with their signaling receptors. However, the significance of TbetaRIII signaling in vivo is not known. In this study, we have sought to determine the role of TbetaRIII during development. We identified the predominant expression sites of TbetaRIII mRNA as liver and heart during midgestation and have disrupted the murine TbetaRIII gene by homologous recombination. Beginning at embryonic day 13.5, mice with mutations in TbetaRIII developed lethal proliferative defects in heart and apoptosis in liver, indicating that TbetaRIII is required during murine somatic development. To assess the effects of the absence of TbetaRIII on the function of its ligands, primary fibroblasts were generated from TbetaRIII-null and wild-type embryos. Our results indicate that TbetaRIII deficiency differentially affects the activities of TGFbeta ligands. Notably, TbetaRIII-null cells exhibited significantly reduced sensitivity to TGFbeta2 in terms of growth inhibition, reporter gene activation, and Smad2 nuclear localization, effects not observed with other ligands. These data indicate that TbetaRIII is an important modulator of TGFbeta2 function in embryonic fibroblasts and that reduced sensitivity to TGFbeta2 may underlie aspects of the TbetaRIII mutant phenotype. PMID- 12773578 TI - Developmentally regulated recruitment of transcription factors and chromatin modification activities to chicken lysozyme cis-regulatory elements in vivo. AB - Expression of the chicken lysozyme gene is upregulated during macrophage differentiation and reaches its highest level in bacterial lipopolysaccharide (LPS)-stimulated macrophages. This is accompanied by complex alterations in chromatin structure. We have previously shown that chromatin fine-structure alterations precede the onset of gene expression in macrophage precursor cells and mark the lysozyme chromatin domain for expression later in development. To further examine this phenomenon and to investigate the basis for the differentiation-dependent alterations of lysozyme chromatin, we studied the recruitment of transcription factors to the lysozyme locus in vivo at different stages of myeloid differentiation. Factor recruitment occurred in several steps. First, early-acting transcription factors such as NF1 and Fli-1 bound to a subset of enhancer elements and recruited CREB-binding protein. LPS stimulation led to an additional recruitment of C/EBPbeta and a significant change in enhancer and promoter structure. Transcription factor recruitment was accompanied by specific changes in histone modification within the lysozyme chromatin domain. Interestingly, we present evidence for a transient interaction of transcription factors with lysozyme chromatin in lysozyme-nonexpressing macrophage precursors, which was accompanied by a partial demethylation of CpG sites. This indicates that a partially accessible chromatin structure of lineage-specific genes is a hallmark of hematopoietic progenitor cells. PMID- 12773579 TI - Accumulation of Fra-1 in ras-transformed cells depends on both transcriptional autoregulation and MEK-dependent posttranslational stabilization. AB - The AP-1 transcription factor plays an essential role in cell proliferation and tumorigenesis. It was previously shown that the fra-1 gene product is upregulated by various oncogenes and is involved in the in vitro and in vivo transformation of thyroid cells. Here we show that the ras oncogene-dependent accumulation of Fra-1 is mediated by a positive feedback mechanism which requires both transcriptional autoregulation and posttranslational stabilization of the protein. The oncogene-dependent transcriptional activation involves the cooperation between both Raf-dependent and Raf-independent pathways and is mediated by an AP-1 site within the fra-1 first intron, which becomes stably occupied by a transcriptionally active Fra-1-containing complex in ras transformed cells. The posttranslational stabilization results in a drastic increase in the Fra-1 half-life in ras-transformed cells and is totally dependent on the activity of the MEK/ERK phosphorylation pathway. The analysis of the Fra-1 transactivation potential shows that the protein is able to stimulate a heterologous promoter in a ras-dependent manner, but the transactivating activity requires the recruitment of a heterodimeric partner. These data show that the alteration of multiple regulatory mechanisms is required for the constitutive activation of Fra-1 as a nuclear target of ras transformation. PMID- 12773580 TI - Perirhinal cortex lesions impair context aversion learning. AB - Rats with perirhinal cortex lesions were compared with sham controls on a conditional discrimination in which saccharin was paired with LiCl in context 1, but paired with saline in context 2. Perirhinal-lesioned rats were slightly slower to acquire the discrimination but reached control levels by the end of acquisition. Both groups showed transfer to familiar tap water, consuming less in context 1 than in context 2. Unlike sham rats, perirhinal rats failed to show an aversion to context 1 on a place choice test. These data provide neuroanatomical support for the postulated difference between Pavlovian conditioning and conditional learning. PMID- 12773581 TI - Sleep deprivation selectively impairs memory consolidation for contextual fear conditioning. AB - Many behavioral and electrophysiological studies in animals and humans have suggested that sleep and circadian rhythms influence memory consolidation. In rodents, hippocampus-dependent memory may be particularly sensitive to sleep deprivation after training, as spatial memory in the Morris water maze is impaired by rapid eye movement sleep deprivation following training. Spatial learning in the Morris water maze, however, requires multiple training trials and performance, as measured by time to reach the hidden platform is influenced by not only spatial learning but also procedural learning. To determine if sleep is important for the consolidation of a single-trial, hippocampus-dependent task, we sleep deprived animals for 0-5 and 5-10 h after training for contextual and cued fear conditioning. We found that sleep deprivation from 0-5 h after training for this task impaired memory consolidation for contextual fear conditioning whereas sleep deprivation from 5-10 h after training had no effect. Sleep deprivation at either time point had no effect on cued fear conditioning, a hippocampus independent task. Previous studies have determined that memory consolidation for fear conditioning is impaired when protein kinase A and protein synthesis inhibitors are administered at the same time as when sleep deprivation is effective, suggesting that sleep deprivation may act by modifying these molecular mechanisms of memory storage. PMID- 12773582 TI - Orbital prefrontal cortex and guidance of instrumental behavior of rats by visuospatial stimuli predicting reward magnitude. AB - The orbital prefrontal cortex (OPFC) is part of a circuitry mediating the perception of reward and the initiation of adaptive behavioral responses. We investigated whether the OPFC is involved in guidance of the speed of instrumental behavior by visuospatial stimuli predictive of different reward magnitudes. Unoperated rats, sham-lesioned rats, and rats with bilateral lesions of the OPFC by N-methyl-D-aspartate (NMDA) were trained in a visuospatial discrimination task. The task required a lever press on the illuminated lever of two available to obtain a food reward. Different reward magnitudes were permanently assigned to lever presses to respective sides of the operant chamber; that is, responses to one lever (e.g., the left one) were always rewarded with one pellet and responses to the other lever with five pellets. On each trial, the position of the illuminated lever was pseudorandomly determined in advance. Results revealed that OPFC lesions did not impair acquisition of the task, as the speed of conditioned responses was significantly shorter with expectancy of a high reward magnitude. In addition, during reversal, shift and reshift of lever position-reward magnitude contingencies and under extinction conditions, performance of the OPFC-lesioned and control groups did not differ. It is concluded that the OPFC in rats might not be critical for adapting behavioral responses to changes of stimulus-reward magnitude contingencies signaled by visuospatial cues. PMID- 12773584 TI - A modified version of the unique cue theory accounts for olfactory compound processing in honeybees. AB - We investigated the capability of honeybees to discriminate between single odorants, binary olfactory mixtures, and ternary olfactory mixtures in olfactory conditioning of the proboscis extension reflex. In Experiment 1, three single odorants (A+, B+, and C+) and three binary mixtures of these odors (AB+, AC+, and BC+) were reinforced while the ternary compound, consisting of all three odors (ABC-), was nonreinforced. In Experiment 2, only one single odorant (A+) and one binary olfactory compound (BC+) were reinforced while the ternary compound (ABC-) consisting of the single odor and the binary compound was nonreinforced. We studied whether bees can solve these problems and whether the course of differentiation can be predicted by the unique cue theory, a modified unique cue theory, or Pearce's configural theory. Honeybees were not able to differentiate reinforced from nonreinforced stimuli in Experiment 1. However, summation to ABC observed at the beginning of training contradicts the predictions of Pearce's configural theory. In Experiment 2, differentiation between the single odorant A and the ternary compound developed more easily than between the binary compound BC and ABC. This pattern of differentiation is in line with a modified unique cue theory and Pearce's configural theory. Summation to ABC at the beginning of training, however, again was at odds with Pearce's configural theory. Thus, olfactory compound processing in honeybees can best be explained by a modified unique cue theory. PMID- 12773583 TI - Effects of environmental enrichment on spatial memory and neurochemistry in middle-aged mice. AB - The present study compared the effects of environmental enrichment on spatial memory, glutamic acid decarboxylase (GAD) activity, and synaptophysin levels in middle-aged male and female mice. Prior to testing, a subset of 18-month-old male and female C57BL/6 mice was housed with two to three toys and a running wheel in the home cage for up to 29 d. Adult mice (7 mo) of both sexes and the remaining middle-aged mice were group (social) housed, but not exposed to enriching objects. After the enrichment period, all mice were tested in a 1-day version of the Morris water maze, in which both spatial and nonspatial memory were assessed. Immediately after testing, the hippocampus and frontoparietal cortex were dissected, and GAD activity and synaptophysin levels were measured. Environmental enrichment reduced the age-related impairment in spatial acquisition and retention; relative to adult social controls, middle-aged enriched mice were unimpaired, whereas middle-aged social controls were impaired. This reduction was similar in middle-aged males and females. Enrichment did not affect cued memory in either sex. Although hippocampal GAD activity was increased by enrichment in males, all other neurochemical measurements were unaffected by enrichment or aging in either sex. These data suggest that environmental enrichment initiated at middle age can reduce age-related impairments in spatial memory in males and females, although the underlying neurobiological mechanisms of this effect remain unknown. PMID- 12773585 TI - Neural correlates of Pavlovian conditioning in components of the neural network supporting ciliary locomotion in Hermissenda. AB - Pavlovian conditioning in Hermissenda consists of pairing light, the conditioned stimulus (CS) with activation of statocyst hair cells, the unconditioned stimulus (US). Conditioning produces CS-elicited foot shortening and inhibition of light elicited locomotion, the two conditioned responses (CRs). Conditioning correlates have been identified in the primary sensory neurons (photoreceptors) of the CS pathway, interneurons that receive monosynaptic input from identified photoreceptors, and putative pedal motor neurons. While cellular mechanisms of acquisition produced by the synaptic interaction between the CS and US pathways are well-documented, little is known about the mechanisms responsible for the generation or expression of the CR. Here we show that in conditioned animals light reduced tonic firing of ciliary activating pedal neurons (VP1) below their pre-CS baseline levels. In contrast, pseudorandom controls expressed a significant increase in CS-elicited tonic firing of VP1 as compared to pre-CS baseline activity. Identified interneurons in the visual pathway that have established polysynaptic connections with VP1 were examined in conditioned animals and pseudorandom controls. Depolarization of identified type Ie interneurons with extrinsic current elicited a significant increase in IPSPs recorded in VP1 pedal neurons of conditioned animals as compared with pseudorandom controls. Conditioning also enhanced intrinsic excitability of type Ie interneurons of conditioned animals as compared to pseudorandom controls. Light evoked a modest increase in IPSP frequency in VP1 of conditioned preparations and a significant decrease in IPSP frequency in VP1 of pseudorandom controls. Our results show that a combination of synaptic facilitation and intrinsic enhanced excitability in identified components of the CS pathway may explain light-elicited inhibition of locomotion in conditioned animals. PMID- 12773586 TI - Olfactory learning in individually assayed Drosophila larvae. AB - Insect and mammalian olfactory systems are strikingly similar. Therefore, Drosophila can be used as a simple model for olfaction and olfactory learning. The brain of adult Drosophila, however, is still complex. We therefore chose to work on the larva with its yet simpler but adult-like olfactory system and provide evidence for olfactory learning in individually assayed Drosophila larvae. We developed a differential conditioning paradigm in which odorants are paired with positive ("+" fructose) or negative ("-" quinine or sodium chloride) gustatory reinforcers. Test performance of individuals from two treatment conditions is compared-one received odorant A with the positive reinforcer and odorant B with a negative reinforcer (A+/B-); animals from the other treatment condition were trained reciprocally (A-/B+). During test, differences in choice between A and B of individuals having undergone either A+/B- or A-/B+ training therefore indicate associative learning. We provide such evidence for both combinations of reinforcers; this was replicable across repetitions, laboratories, and experimenters. We further show that breaks improve performance, in accord with basic principles of associative learning. The present individual assay will facilitate electrophysiological studies, which necessarily use individuals. As such approaches are established for the larval neuromuscular synapse, but not in adults, an individual larval learning paradigm will serve to link behavioral levels of analysis to synaptic physiology. PMID- 12773588 TI - Chronic complex depression. PMID- 12773587 TI - Analysis of probabilistic classification learning in patients with Parkinson's disease before and after pallidotomy surgery. AB - This study examined the characteristics of probabilistic classification learning, a form of implicit learning previously shown to be impaired in patients with basal ganglia dysfunction (e.g., Parkinson's disease). In this task, subjects learn to predict the weather using associations that are formed gradually across many trials, because of the probabilistic nature of the cue-outcome relationships. Patients with Parkinson's disease, both before and after pallidotomy, and age-matched control subjects, exhibited evidence of probabilistic classification learning across 100 training trials. However, pallidotomy appears to hinder the learning of associations most implicit in nature (i.e., weakly associated cues). Although subjects were most sensitive to single-cue associations when learning the task, there is evidence that cue combinations contribute significantly to probability learning. The utility of multiple dependent measures is discussed. PMID- 12773589 TI - APA issues blueprint for mental health system in advance of presidential commission's report. PMID- 12773590 TI - Datapoints: perceived benefits of substance abuse treatments. PMID- 12773591 TI - Characteristics of patients referred by police to a psychiatric emergency service. PMID- 12773592 TI - La-la land meets DSM-IV: the pleasures and pitfalls of celluloid diagnostics. PMID- 12773593 TI - Health plan options at fortune 100 companies: available coverage for mental health care consumers. PMID- 12773594 TI - A VA inpatient respite program for patients with dementia. PMID- 12773595 TI - On confronting myself and the world. PMID- 12773596 TI - The five-site health and risk study of blood-borne infections among persons with severe mental illness. AB - This article outlines the history and rationale of a multisite study of blood borne infections among persons with severe mental illness reported in this special section of Psychiatric Services. The general problem of blood-borne diseases in the United States is reviewed, particularly as it affects people with severe mental illness and those with comorbid substance use disorders. The epidemiology and natural history of three of the most important infections are reviewed: the human immunodeficiency virus (HIV), the hepatitis B virus, and the hepatitis C virus. Current knowledge about blood-borne diseases among people with severe mental illness as well as information on current treatment advances for hepatitis C are summarized. A heuristic model, based on the pragmatic, empirical, and conceptual issues that influenced the final study design, is presented. The specific rationale of the five-site collaborative design is discussed, as well as the sampling frames, measures, and procedures used at the participating sites. Alternative strategies for analyzing data deriving from multisite studies that use nonrandomized designs are described and compared. Finally, each of the articles in this special section is briefly outlined, with reference to the overall hypotheses of the studies. PMID- 12773597 TI - Risk factors for HIV, hepatitis B, and hepatitis C among persons with severe mental illness. AB - OBJECTIVE: Previous reports have indicated that persons with severe mental illness have an elevated risk of contracting HIV, hepatitis B, and hepatitis C compared with the general population. This study extends earlier findings by examining the factors that are most predictive of serologic status among persons with severe mental illness. METHOD: S: A total of 969 persons with severe mental illness from five sites in four states were approached to take part in an assessment involving testing for blood-borne infections and a one-time standardized interview containing questions about sociodemographic characteristics, substance use, risk behaviors for sexually transmitted diseases, history of sexually transmitted diseases, and health care. RESULTS: The greater the number of risk behaviors, the greater was the likelihood of infection, both for persons in more rural locations (New Hampshire and North Carolina), where the prevalence of infection was lower, and those in urban locations (Hartford, Connecticut; Bridgeport, Connecticut; and Baltimore, Maryland), where the prevalence was higher. Although no evidence was found that certain behaviors increase a person's risk of one blood-borne infection while other behaviors increase the risk of a different infection, it is conceivable that more powerful research designs would reveal some significant differences among the risks. CONCLUSION: S: Clinicians should be attentive to these risk factors so as to encourage appropriate testing, counseling, and treatment. PMID- 12773598 TI - Substance abuse and the transmission of hepatitis C among persons with severe mental illness. AB - OBJECTIVES: The authors sought to better understand the relationship of substance abuse to higher rates of transmission of hepatitis C among persons with severe mental illness. METHOD: S: The authors assessed 668 persons with severe mental illness for HIV, hepatitis B, and hepatitis C infection through venipuncture. Demographic characteristics, substance abuse, and risk behaviors for blood-borne infections were assessed through interviews and collection of clinical data. RESULTS: Eighty-two percent of the assessed persons were not infected, and 18 percent had hepatitis C. Among those with hepatitis C infection, 546 (82 percent) tested negative for all viruses. Of the 122 (18 percent) who had hepatitis C, 53 (8 percent) had only hepatitis C, 56 (8 percent) had both hepatitis C and hepatitis B, three (1 percent) had hepatitis C and HIV, and ten (2 percent) had all three infections. More than 20 percent of the sample reported lifetime intravenous drug use, and 14 percent reported lifetime needle sharing. Fifty seven percent had sniffed of snorted cocaine, and 39 percent had smoked crack. A stepwise regression model was used to identify interaction effects of these behaviors and risk of hepatitis C infection among persons with severe mental illness. Use of needles and of crack cocaine were associated with a large increase in the likelihood of hepatitis C infection. CONCLUSION: S: The high rates of co-occurring substance use disorders among persons with severe mental illness, coupled with the role of substance abuse as the primary vector for hepatitis C transmission, warrants special consideration. PMID- 12773599 TI - Gender differences in hepatitis C infection and risks among persons with severe mental illness. AB - OBJECTIVES: The authors assessed gender differences in hepatitis C infection and associated risk behaviors among persons with severe mental illness. METHOD: S: The sample consisted of 777 patients (251 women and 526 men) from four sites. RESULTS: Across sites, the rate of hepatitis C infection among men was nearly twice that among women. Clear differences were noted in hepatitis C risk behaviors. Men had higher rates of lifetime drug-related risk behaviors: needle use (23.1 percent compared with 12.5 percent), needle sharing (17.6 percent compared with 7.7 percent), and crack cocaine use (45.2 percent compared with 30.8 percent). Women had significantly higher rates of lifetime sexual risk behaviors: unprotected sex in exchange for drugs (17.8 percent compared with 11.2 percent), unprotected sex in exchange for money or gifts (30.6 percent compared with 17 percent), unprotected vaginal sex (94 percent compared with 89.7 percent), and anal sex (33.7 percent compared with 22.6 percent). Gender appeared to modify some sex risks. Unprotected sex in exchange for drugs increased the risk of hepatitis C seropositivity for both men and women. In the multivariate model, gender was not significantly associated with hepatitis C seropositivity after adjustment for other risk factors. CONCLUSION: S: Gender differences in the lifetime rates of drug risks explain the higher rates of hepatitis C infection among men with severe mental illness. PMID- 12773600 TI - Regular sources of medical care among persons with severe mental illness at risk of hepatitis C infection. AB - OBJECTIVE: An estimated 19.6 percent of persons with severe mental illness are infected with the hepatitis C virus. Given the pressing need to identify and treat persons with severe mental illness who are at risk of hepatitis C infection and transmission, the authors sought to estimate the proportion of hepatitis C positive and -negative persons with severe mental illness who have a regular source of medical care. METHOD: S: Data for this study were obtained from 777 adults with severe mental illness at four diverse geographic sites at which respondents with severe mental illness participated in a structured interview and laboratory testing for HIV infection, AIDS, hepatitis B infection, and hepatitis C infection. RESULTS: In bivariate analyses, 54.2 percent of hepatitis C-positive and 62.5 percent of hepatitis C-negative study participants with severe mental illness had a regular source of medical care. In multivariate analyses in which potential confounders were statistically controlled for, hepatitis C-positive persons with severe mental illness were less than half as likely as hepatitis C negative persons to have a regular source of care. Being older, married, insured, or employed or having self-reported health problems increased the likelihood of receiving care. Being black or male or living in a community with high exposure to community violence lowered those odds. CONCLUSION: There is an urgent need to improve access to medical care for persons with severe mental illness, especially those who may be at high risk of or are already infected with the hepatitis C virus. PMID- 12773601 TI - Responding to blood-borne infections among persons with severe mental illness. AB - The Five-Site Health and Risk Study estimated prevalence rates of blood-borne infections, including the human immunodeficiency virus (HIV), hepatitis B, and hepatitis C, and addressed risk factors and correlates of infection among persons with severe mental illness. In this final article of the special section in this issue of Psychiatric Services, the authors review public health recommendations and best practices and discuss the implications of these results for community mental health care of clients with severe mental illness. Standard public health recommendations could be modified for use by community mental health providers. In addition, expansion of integrated dual disorders treatments and improving linkage with specialty medical care providers are recommended. PMID- 12773602 TI - Ethical exploration of the least restrictive alternative. AB - Although there have been critiques about the application of the "least restrictive alternative" to mental health policies in the past three decades, no critical analysis of the ethical logic of this principle has been put forward. The author explores three main ethical theories-liberalism, utilitarianism, and communitarianism-and explains how liberalism and subjective utilitarianism, which evidently uphold the least restrictive alternative in the name of individual rights and preference for liberty, effectively disenfranchise patients and members of their family. Contrarily, objective utilitarianism, with its highlight on cost-effectiveness analysis, would actually lay solid ground for mental health policies that attend not simply to ideological belief but rather to the question of which treatments can most cost-effectively meet the complex needs of patients. In addition, under communitarianism the evidence derived from cost-effectiveness analysis can be used to reinterpret the traditional value in mental health policies and thereby lead to the policy in favor of the most cost-effective alternative. PMID- 12773603 TI - Patterns and predictors of service use and unmet needs among aging families of adults with severe mental illness. AB - OBJECTIVE: This study examined patterns and predictors of use of and unmet need for support services among aging families of adults with severe mental illness by using an expanded version of the Andersen Behavioral Model. METHOD: S: Mailed surveys were completed by 157 mothers from 41 states who lived with and provided care to adult offspring with serious mental disorders, primarily schizophrenia or schizoaffective disorders. The mean age of the mothers was 67 years (range, 50 to 88 years). RESULTS: Although unique patterns were observed among individual services, overall service use was low despite high perceived need for services. The greatest unmet needs were for social or recreational programming, training in behavior management, and information on planning for the future. Regression analyses based on the expanded Andersen model revealed that greater service use occurred when offspring spent their days away from home, mothers received higher levels of informal support, and offspring were in poorer physical health. Greater unmet needs for services were reported when mothers experienced higher burden, perceived more age-related changes in themselves, and had offspring who typically spent their days at home. CONCLUSION: S: The needs and resources of the entire family, including access to informal social support, should be considered in attempts to identify predictors of the use of and need for services among persons with chronic and severe mental illness. The findings of this study also point to the need for family education in how to locate community services as well as for better and more sensitive community services intended for the entire aging family. PMID- 12773604 TI - Mental health services received by depressed persons who visited general practitioners and family doctors. AB - OBJECTIVES: This study estimated the rates of mental health service provision and of specialist referral in primary care in Canada and investigated factors associated with receiving mental health services and with referral to mental health specialists among persons who reported major depressive episodes. METHOD: S: Data from the 1998-1999 Canadian National Population Health Survey were used. The 608 respondents who reported having major depressive episodes in the 12 months preceding the survey and who reported contacting a general practitioner or family doctor during that time were included in the study. The rates of provision of mental health services by general practitioners and family doctors and of referral to mental health specialists were calculated. Demographic, socioeconomic, and clinical characteristics associated with receiving mental health services and with referral to specialists were investigated. RESULTS: Among the 608 respondents who had contacted general practitioners or family doctors for any reason, 153 had contacted them for emotional or mental problems. Of this subgroup of 153, 64.5 percent received mental health services either from these practitioners or by referral to specialists, and 26 percent were referred to mental health specialists. Depressed respondents who reported having talked to a general practitioner or family doctor about mental health problems, who reported impairment, and whose depressive symptoms had lasted eight or more weeks were more likely to have received mental health services. Respondents aged 12 to 24 years were more likely to be referred to mental health specialists. CONCLUSION: S: Impairment associated with depression and chronicity of depressive symptoms appear to be the primary determinants of the decisions made by general practitioners and family doctors about providing mental health services. Patients' willingness to consult with general practitioners or family doctors for mental health problems may also be a key factor, both for effective management of depression in primary care settings and for referral to mental health specialists. PMID- 12773605 TI - Cost-effectiveness of critical time intervention to reduce homelessness among persons with mental illness. AB - OBJECTIVES: Cost-effective programs are needed to assist homeless persons with severe mental illness in their transition from shelters to community living. The authors investigated the cost-effectiveness of the critical time intervention program, a time-limited adaptation of intensive case management, which has been shown to significantly reduce recurrent homelessness among men with severe mental illness. METHOD: S: Ninety-six study participants recruited from a psychiatric program in a men's public shelter from 1991 to 1993 were randomly assigned to the critical time intervention program or to usual services. Costs and housing outcomes for the two groups were examined over 18 months. RESULTS: Over the study period, the critical time intervention group and the usual services group incurred mean costs of 52,374 dollars and 51,649 dollars, respectively, for acute care services, outpatient services, housing and shelter services, criminal justice services, and transfer income. During the same period, the critical time intervention group experienced significantly fewer homeless nights than the usual care group (32 nights versus 90 nights). For each willingness-to-pay value--the additional price society is willing to spend for an additional nonhomeless night- greater than 152 dollars, the critical time intervention group exhibited a significantly greater net housing stability benefit, indicating cost effectiveness, compared with usual care. CONCLUSION: S: Although difficult to conduct, studies of the cost-effectiveness of community mental health programs can yield rich information for policy makers and program planners. The critical time intervention program is not only an effective method to reduce recurrent homelessness among persons with severe mental illness but also represents a cost effective alternative to the status quo. PMID- 12773606 TI - Detection of illicit substance use among persons with schizophrenia by radioimmunoassay of hair. AB - OBJECTIVE: Illicit substance use is a potent risk factor for poor outcomes in schizophrenia, yet methods for detecting substance use consistently underestimate the problem. The purpose of this study was to assess whether use of a relatively new method of detection, radioimmunoassay of hair, improved detection and was acceptable to patients with serious mental illness. METHOD: S: Persons already participating in a longitudinal naturalistic study of schizophrenia treatment were approached for participation in this study. The 203 persons who consented were interviewed and submitted urine and hair samples for laboratory measures of potential substances of abuse. Radioimmunoassay of hair was used to detect the use of amphetamines, cocaine, marijuana, opiates, and phencyclidine (PCP) in the preceding three months. RESULTS: Of the 203 participants, only 33 (16.3 percent) self-reported illicit substance use, and only 25 (12.4 percent) had a positive urine test, but 63 (31.0 percent) had a positive hair assay. When all detection methods were combined--self-report, urine test, and hair assay--78 participants (38.4 percent) were classified as users in the preceding three months. Few of those asked to participate (20, or 9.9 percent) refused hair analysis. CONCLUSION: S: Radioimmunoassay of hair appears to be a promising method for improving assessment of illicit substance use among persons with schizophrenia. Most participants appeared to find hair analysis an acceptable procedure, although this conclusion warrants further study. The test's three-month window of detection may make it a valuable method for assessing and monitoring use over time. PMID- 12773607 TI - Psychiatry's remarkable journey: the past 40 years. PMID- 12773608 TI - A comparison of gynecological variables and service use among older women with and without schizophrenia. AB - Gynecological variables and service use were examined in a sample of 65 older women outpatients who had schizophrenia and in a control group of 51 women who were not known to have a psychiatric diagnosis. The groups were similar in their age at menarche and at menopause, use of oral contraceptives, and number of pregnancies and births. The groups differed in their receipt of several gynecological services; women with schizophrenia were less likely to have received mammograms or pelvic examinations and Pap tests or to have ever been prescribed hormone replacement therapy. These results suggest that women with schizophrenia receive fewer gynecological services than other women. Interventions at the patient, provider, and system levels may be needed to address this disparity in service use. PMID- 12773609 TI - The effect of housing interventions on neuropsychological functioning among homeless persons with mental illness. AB - The authors tested the hypotheses that neuropsychological functioning would improve after homeless persons with severe and persistent mental illness were provided with housing and that executive functioning would improve more among those placed in group homes than among those placed in independent apartments. A total of 114 persons with serious and persistent mental illness who were stable residents of homeless shelters completed neuropsychological testing and were randomly assigned to group homes or independent apartments; 91 participants (52 assigned to group homes and 39 assigned to independent apartments) were retested after 18 months. Overall neuropsychological functioning improved significantly across the full study sample. Executive performance, measured by the Wisconsin Card Sorting Test, decreased significantly among persons assigned to independent apartments and increased, but not significantly, among those assigned to group homes. The findings suggest that providing housing for persons who have severe and persistent mental illness improves cognitive functioning but that independent living may diminish executive functioning. PMID- 12773611 TI - Home care nursing and detection of depression. PMID- 12773612 TI - Clinical care of veterans seeking compensation. PMID- 12773613 TI - Working with people who have mental retardation. PMID- 12773614 TI - GAO finds inappropriate child placements in welfare, juvenile justice systems to obtain mental health care. PMID- 12773616 TI - Mapping the genetic variation of executive attention onto brain activity. AB - Brain imaging data have repeatedly shown that the anterior cingulate cortex is an important node in the brain network mediating conflict. We previously reported that polymorphisms in dopamine receptor (DRD4) and monoamine oxidase A (MAOA) genes showed significant associations with efficiency of handling conflict as measured by reaction time differences in the Attention Network Test (ANT). To examine whether this genetic variation might contribute to differences in brain activation within the anterior cingulate cortex, we genotyped 16 subjects for the DRD4 and MAOA genes who had been scanned during the ANT. In each of the two genes previously associated with more efficient handling of conflict in reaction time experiments, we found a polymorphism in which persons with the allele associated with better behavioral performance showed significantly more activation in the anterior cingulate while performing the ANT than those with the allele associated with worse performance. The results demonstrate how genetic differences among individuals can be linked to individual differences in neuromodulators and in the efficiency of the operation of an appropriate attentional network. PMID- 12773615 TI - Redirection of B cell responsiveness by transforming growth factor beta receptor. AB - The multifunctional transforming growth factor beta receptor (TbetaR) ligand pair plays a central role in the regulation of lymphocyte homeostasis and prevention of autoimmunity. Although the mechanisms underlying the induction of transcriptional modulators by TbetaR have been studied in considerable detail, relatively little is known about the regulatory pathways targeted. To shed light on the mechanisms involved in negative regulation of B cell responses we identified TbetaR-dependent transcriptome changes by comparative gene expression profiling of normal and TbetaR-deficient primary B cells. The data reveal TbetaR mediated induction of inhibitors of antigen receptor signaling (Ship-1, CD72) as well as inhibitors of the Jak/Stat pathway and signaling by means of Toll-like receptors (SOCS1,3). These inhibitory effects are complemented by induction of antiproliferative transcription factors. In contrast to this inhibition, G protein-coupled receptors such as CXCR4 and agonists mediating Ca2+ flux (inositol trisphosphate receptor subtype 2) are induced by TbetaR, indicating enhancement of the Ca2+ storage/ release system and chemotactic responses. Suppression of proapoptotic genes suggests support of cell survival. Confirming the shift in B cell responsiveness, antigen-receptor-mediated activation of Syk and phospholipase C-gamma2, as well as Stat6 phosphorylation, is inhibited, whereas chemotaxis, Ca2+ release, and cell survival are enhanced in transforming growth factor-beta-sensitive B cells. The data provide a molecular basis for TbetaR-mediated inhibition of B cell responsiveness and indicate that TbetaR maintains homeostasis not only through inhibition of the cell cycle but also by delivering a coherent instructive signal that redirects responsiveness to microenvironmental cues. PMID- 12773617 TI - Competition and representation during memory retrieval: roles of the prefrontal cortex and the posterior parietal cortex. AB - In this functional-MRI study we examined the hypothesis that the prefrontal cortex responds differently to the extent of competition during retrieval, whereas the parietal cortex is responsible for problem representation that should not be directly related to the competition. Participants mastered arbitrary person-location pairs, and their recognition memory was tested in a functional MRI session. The pairs were constructed such that a person was associated with one, two, or three different locations and vice versa. The recognition time increased with the number of associations, reflecting increased competition. A confirmatory analysis of imaging data with prespecified prefrontal and parietal regions showed that, although both regions were highly involved during memory retrieval, only the prefrontal region responded to the levels of competition. This result was consistent with predictions of an information-processing model as well as with an exploratory identification of regions of interest. PMID- 12773618 TI - Direct observation of photolysis-induced tertiary structural changes in hemoglobin. AB - Human Hb, an alpha2beta2 tetrameric oxygen transport protein that switches from a T (tense) to an R (relaxed) quaternary structure during oxygenation, has long served as a model for studying protein allostery in general. Time-resolved spectroscopic measurements after photodissociation of CO-liganded Hb have played a central role in exploring both protein dynamical responses and molecular cooperativity, but the direct visualization and the structural consequences of photodeligation have not yet been reported. Here we present an x-ray study of structural changes induced by photodissociation of half-liganded T-state and fully liganded R-state human Hb at cryogenic temperatures (25-35 K). On photodissociation of CO, structural changes involving the heme and the F-helix are more marked in the alpha subunit than in the beta subunit, and more subtle in the R state than in the T state. Photodeligation causes a significant sliding motion of the T-state beta heme. Our results establish that the structural basis of the low affinity of the T state is radically different between the subunits, because of differences in the packing and chemical tension at the hemes. PMID- 12773619 TI - A unique mechanism for protein processing and degradation in Arabidopsis thaliana. AB - Precursor protease vesicles are plant-specific compartments containing precursors of enzymes that are thought to participate in the degradation of cellular components in organs undergoing senescence. We report in vivo evidence that the precursor protease vesicle-localized vacuolar processing enzyme-gamma (VPEgamma) is critical for maturation of the plant vacuolar protease AtCPY. We also provide biochemical and functional evidence that VPEgamma is involved in degradation of the vacuolar invertase AtFruct4 in aging tissues. Moreover, a proteomics-based approach identified various proteins found in the vacuoles of aging vpegamma mutants but not in WT plants, suggesting a unique role of VPEgamma in protein processing and degradation in Arabidopsis. PMID- 12773620 TI - Decorin-binding proteins A and B confer distinct mammalian cell type-specific attachment by Borrelia burgdorferi, the Lyme disease spirochete. AB - Host cell binding is an essential step in colonization by many bacterial pathogens, and the Lyme disease agent, Borrelia burgdorferi, which colonizes multiple tissues, is capable of attachment to diverse cell types. Glycosaminoglycans (GAGs) are ubiquitously expressed on mammalian cells and are recognized by multiple B. burgdorferi surface proteins. We previously showed that B. burgdorferi strains differ in the particular spectrum of GAGs that they recognize, leading to differences in the cultured mammalian cell types that they efficiently bind. The molecular basis of these binding specificities remains undefined, due to the difficulty of analyzing multiple, potentially redundant cell attachment pathways and to the paucity of genetic tools for this pathogen. In the current study, we show that the expression of decorin-binding protein (Dbp) A and/or DbpB, two B. burgdorferi surface proteins that bind GAGs, is sufficient to convert a high-passage nonadherent B. burgdorferi strain into one that efficiently binds 293 epithelial cells. Epithelial cell attachment was mediated by dermatan sulfate, and, consistent with this GAG-binding specificity, these recombinant strains did not bind EA-Hy926 endothelial cells. The GAG binding properties of bacteria expressing DbpB or DbpA were distinguishable, and DbpB but not DbpA promoted spirochetal attachment to C6 glial cells. Thus, DbpA and DbpB may each play central but distinct roles in cell type-specific binding by Lyme disease spirochetes. This study illustrates that transformation of high passage B. burgdorferi strains may provide a relatively simple genetic approach to analyze virulence-associated phenotypes conferred by multiple bacterial factors. PMID- 12773621 TI - Ligand dynamics in a protein internal cavity. AB - We have studied the temperature dependence of the IR stretch bands of carbon monoxide (CO) in the Xe 4 internal cavity of myoglobin mutant L29W-S108L at cryogenic temperatures. Pronounced changes of band areas and positions were analyzed quantitatively by using a simple dynamic model in which CO rotation in the cavity is constrained by a static potential. The librational dynamics of the CO causes a decrease of the total spectral area. A strong local electric field splits the CO stretch absorption into a doublet, indicating that CO can assume opposite orientations in the cavity. With increasing temperature, the two peaks approach each other, because the average angle of the CO with respect to the electric field increases. A combined classical and quantum-mechanical analysis precisely reproduces the observed temperature dependencies of both spectral area and peak shifts. It yields the height of the energy barrier between the two wells associated with opposite CO orientations, V0 approximately 2 kJ/mol, and the frequency of oscillation within a well, omega approximately 25 cm(-1). The electric field in the protein cavity was estimated as 10 MV/cm. PMID- 12773622 TI - Protein structure change studied by hydrogen-deuterium exchange, functional labeling, and mass spectrometry. AB - An automated high-throughput, high-resolution deuterium exchange HPLC-MS method (DXMS) was used to extend previous hydrogen exchange studies on the position and energetic role of regulatory structure changes in hemoglobin. The results match earlier highly accurate but much more limited tritium exchange results, extend the analysis to the entire sequence of both hemoglobin subunits, and identify some energetically important changes. Allosterically sensitive amide hydrogens located at near amino acid resolution help to confirm the reality of local unfolding reactions and their use to evaluate resolved structure changes in terms of allosteric free energy. PMID- 12773623 TI - The hSK4 (KCNN4) isoform is the Ca2+-activated K+ channel (Gardos channel) in human red blood cells. AB - The question is, does the isoform hSK4, also designated KCNN4, represent the small conductance, Ca2+-activated K+ channel (Gardos channel) in human red blood cells? We have analyzed human reticulocyte RNA by RT-PCR, and, of the four isoforms of SK channels known, only SK4 was found. Northern blot analysis of purified and synchronously growing human erythroid progenitor cells, differentiating from erythroblasts to reticulocytes, again showed only the presence of SK4. Western blot analysis, with an anti-SK4 antibody, showed that human erythroid progenitor cells and, importantly, mature human red blood cell ghost membranes, both expressed the SK4 protein. The Gardos channel is known to turn on, given inside Ca2+, in the presence but not the absence of external Ko+ and remains refractory to Ko+ added after exposure to inside Ca2+. Heterologously expressed SK4, but not SK3, also shows this behavior. In inside-out patches of red cell membranes, the open probability (Po) of the Gardos channel is markedly reduced when the temperature is raised from 27 to 37 degrees C. Net K+ efflux of intact red cells is also reduced by increasing temperature, as are the Po values of inside-out patches of Chinese hamster ovary cells expressing SK4 (but not SK3). Thus the envelope of evidence indicates that SK4 is the gene that codes for the Gardos channel in human red blood cells. This channel is important pathophysiologically, because it represents the major pathway for cell shrinkage via KCl and water loss that occurs in sickle cell disease. PMID- 12773625 TI - Rapid biosynthesis of N-linolenoyl-L-glutamine, an elicitor of plant volatiles, by membrane-associated enzyme(s) in Manduca sexta. AB - In response to elicitors in the oral secretions of caterpillars, plants produce and release volatile chemicals that attract predators and parasitoids of the caterpillar while it feeds. The most prevalent elicitors are fatty acid amides consisting of 18-carbon polyunsaturated fatty acids coupled with l-glutamine. We demonstrate rapid CoA- and ATP-independent in vitro biosynthesis of the fatty acid amide elicitor, N-linolenoyl-l-glutamine, by microsomal fractions of several alimentary tissues in Manduca sexta. N-linolenoyl-l-glutamine is a structural analog of several other elicitors including volicitin, the first fatty acid amide elicitor identified in caterpillars. The enzyme(s) that catalyzed biosynthesis of N-linolenoyl-l-glutamine was localized within the integral membrane protein fraction extracted from microsomes by Triton X-114 detergent phase partitioning and had maximum activity at alkaline pH. We found no evidence suggesting microbial or tissue-independent biosynthesis of N-linolenoyl-l-glutamine in M. sexta. The in vitro biosynthesis of N-linolenoyl-l-glutamine by membrane associated enzyme(s) in M. sexta represents direct evidence of fatty acid amide synthesis by caterpillar tissues. PMID- 12773624 TI - Aberrant lamination in the cerebral cortex of mouse embryos lacking DNA topoisomerase IIbeta. AB - We have examined corticogenesis in mouse embryos lacking DNA topoisomerase IIbeta (IIbeta) in the brain or in all tissues. The absence of IIbeta, a type II DNA topoisomerase normally expressed in postmitotic cells in the developing cortex, severely affects cerebral stratification: no subplate is discernible, and neurons born at later stages of corticogenesis fail to migrate to the superficial layers. This abnormal pattern of neuron positioning in the cerebral cortex is reminiscent of that observed in mouse mutants defective in the reelin-signaling pathway. Significantly, the level of reelin in the neocortex is much reduced when IIbeta is absent. These results implicate a role of IIbeta in brain development. The enzyme may be required in implementing particular genetic programs in postmitotic cells, such as reelin expression in Cajal-Retzius cells, perhaps through its action on nucleoprotein structure of particular chromosomal regions. PMID- 12773626 TI - A major role for the TATA box in recruitment of chromatin modifying complexes to a globin gene promoter. AB - The developmentally regulated mammalian beta-globin genes are activated by a distant locus control region/enhancer. To understand the role of chromatin remodeling complexes in this activation, we used stably replicated chromatin templates, in which transcription activation of the human embryonic epsilon globin gene depends on the tandem Maf-recognition elements (MAREs) within the beta-globin locus control region HS2 enhancer, to which the erythroid factor NF E2 binds. The HS2 MAREs are required for nucleosome mobilization and histone hyperacetylation at the distant promoter. Nucleosome mobilization also requires the promoter TATA box, and is independent of histone hyperacetylation. In contrast, promoter hyperacetylation requires the promoter GATA-1, and CACC-factor activator motifs, as well as the TATA box. ChIP analysis reveals that NF-E2 is associated with the active epsilon-globin promoter, which lacks an NF-E2 binding sequence, in a TATA box and HS2/MARE-dependent fashion. NF-E2 association with the epsilon-globin promoter coincides with that of RNA polymerase II at both regulatory sites. The results emphasize MARE-TATA box interactions in the recruitment of complexes modifying promoter chromatin for transcription activation and imply close physical interaction between widely separated regulatory sequences mediated through these sites. PMID- 12773627 TI - Cu/Zn superoxide dismutase mutants associated with amyotrophic lateral sclerosis show enhanced formation of aggregates in vitro. AB - Mutations in Cu/Zn superoxide dismutase (SOD) are associated with the fatal neurodegenerative disorder amyotrophic lateral sclerosis (ALS). There is considerable evidence that mutant SOD has a gain of toxic function; however, the mechanism of this toxicity is not known. We report here that purified SOD forms aggregates in vitro under destabilizing solution conditions by a process involving a transition from small amorphous species to fibrils. The assembly process and the tinctorial and structural properties of the in vitro aggregates resemble those for aggregates observed in vivo. Furthermore, the familial ALS SOD mutations A4V, G93A, G93R, and E100G decrease protein stability, which correlates with an increase in the propensity of the mutants to form aggregates. These mutations also increase the rate of protein unfolding. Our results suggest three possible mechanisms for the increase in aggregation: (i) an increase in the equilibrium population of unfolded or of partially unfolded states, (ii) an increase in the rate of unfolding, and (iii) a decrease in the rate of folding. Our data support the hypothesis that the gain of toxic function for many different familial ALS-associated mutant SODs is a consequence of protein destabilization, which leads to an increase in the formation of cytotoxic protein aggregates. PMID- 12773628 TI - Gender differences in the membrane transport of endogenous and exogenous compounds. AB - Gender differences have been well described in pharmacokinetics and contribute to the interindividual variation in drug disposition, therapeutic response, and drug toxicity. Sex-related differences in the membrane transport of endogenous substrates and xenobiotics have been reported in various organs of the body including kidney, liver, intestine, and brain. These gender-related differences in transport systems could also contribute to interindividual variability in pharmacokinetics and pharmacodynamics. This review will focus on current knowledge of gender-associated differences in the transport of endogenous and exogenous compounds in a variety of body organs and will discuss the implications and the clinical significance of these observations. PMID- 12773629 TI - Interleukin-10 therapy--review of a new approach. AB - Interleukin (IL)-10 is an important immunoregulatory cytokine produced by many cell populations. Its main biological function seems to be the limitation and termination of inflammatory responses and the regulation of differentiation and proliferation of several immune cells such as T cells, B cells, natural killer cells, antigen-presenting cells, mast cells, and granulocytes. However, very recent data suggest IL-10 also mediates immunostimulatory properties that help to eliminate infectious and noninfectious particles with limited inflammation. Numerous investigations, including expression analyses in patients, in vitro and animal experiments suggest a major impact of IL-10 in inflammatory, malignant, and autoimmune diseases. So IL-10 overexpression was found in certain tumors as melanoma and several lymphomas and is considered to promote further tumor development. Systemic IL-10 release is a powerful tool of the central nervous system to prevent hyperinflammatory processes by activation of the neuro endocrine axis following acute stress reactions. In contrast, a relative IL-10 deficiency has been observed and is regarded to be of pathophysiological relevance in certain inflammatory disorders characterized by a type 1 cytokine pattern such as psoriasis. Recombinant human IL-10 has been produced and is currently being tested in clinical trials. This includes rheumatoid arthritis, inflammatory bowel disease, psoriasis, organ transplantation, and chronic hepatitis C. The results are heterogeneous. They give new insight into the immunobiology of IL-10 and suggest that the IL-10/IL-10 receptor system may become a new therapeutic target. PMID- 12773630 TI - The pharmacology of nitric oxide in the peripheral nervous system of blood vessels. AB - Unanticipated, novel hypothesis on nitric oxide (NO) radical, an inorganic, labile, gaseous molecule, as a neurotransmitter first appeared in late 1989 and into the early 1990s, and solid evidences supporting this idea have been accumulated during the last decade of the 20th century. The discovery of nitrergic innervation of vascular smooth muscle has led to a new understanding of the neurogenic control of vascular function. Physiological roles of the nitrergic nerve in vascular smooth muscle include the dominant vasodilator control of cerebral and ocular arteries, the reciprocal regulation with the adrenergic vasoconstrictor nerve in other arteries and veins, and in the initiation and maintenance of penile erection in association with smooth muscle relaxation of the corpus cavernosum. The discovery of autonomic efferent nerves in which NO plays key roles as a neurotransmitter in blood vessels, the physiological roles of this nerve in the control of smooth muscle tone of the artery, vein, and corpus cavernosum, and pharmacological and pathological implications of neurogenic NO have been reviewed. This nerve is a postganglionic parasympathetic nerve. Mechanical responses to stimulation of the nerve, mainly mediated by NO, clearly differ from those to cholinergic nerve stimulation. The naming "nitrergic or nitroxidergic" is therefore proposed to avoid confusion of the term "cholinergic nerve", from which acetylcholine is released as a major neurotransmitter. By establishing functional roles of nitrergic, cholinergic, adrenergic, and other autonomic efferent nerves in the regulation of vascular tone and the interactions of these nerves in vivo, especially in humans, progress in the understanding of cardiovascular dysfunctions and the development of pharmacotherapeutic strategies would be expected in the future. PMID- 12773631 TI - Generation of the melatonin endocrine message in mammals: a review of the complex regulation of melatonin synthesis by norepinephrine, peptides, and other pineal transmitters. AB - Melatonin, the major hormone produced by the pineal gland, displays characteristic daily and seasonal patterns of secretion. These robust and predictable rhythms in circulating melatonin are strong synchronizers for the expression of numerous physiological processes in photoperiodic species. In mammals, the nighttime production of melatonin is mainly driven by the circadian clock, situated in the suprachiasmatic nucleus of the hypothalamus, which controls the release of norepinephrine from the dense pineal sympathetic afferents. The pivotal role of norepinephrine in the nocturnal stimulation of melatonin synthesis has been extensively dissected at the cellular and molecular levels. Besides the noradrenergic input, the presence of numerous other transmitters originating from various sources has been reported in the pineal gland. Many of these are neuropeptides and appear to contribute to the regulation of melatonin synthesis by modulating the effects of norepinephrine on pineal biochemistry. The aim of this review is firstly to update our knowledge of the cellular and molecular events underlying the noradrenergic control of melatonin synthesis; and secondly to gather together early and recent data on the effects of the nonadrenergic transmitters on modulation of melatonin synthesis. This information reveals the variety of inputs that can be integrated by the pineal gland; what elements are crucial to deliver the very precise timing information to the organism. This also clarifies the role of these various inputs in the seasonal variation of melatonin synthesis and their subsequent physiological function. PMID- 12773633 TI - Introduction of Wx transgene into rice wx mutants leads to both high- and low amylose rice. AB - The Waxy (Wx) gene encodes a granule-bound starch synthase (GBSS) that plays a key role in the amylose synthesis of rice and other plant species. Two functional Wx alleles of rice exist: Wx(a), which produces a large amount of amylose, and Wx(b), which produces a smaller amount of amylose because of the mutation at the 5' splice site of intron 1. Wx(b) is largely distributed in Japonica cultivars, and high amylose cultivars do not exist in Japonica cultivars. We introduced the cloned Wx(a) cDNA into null-mutant Japonica rice (wx). The amylose contents of these transgenic plants were 6-11% higher than that of the original cultivar, Labelle, which carries the Wx(a) allele, although the levels of the Wx protein in the transgenic rice were equal to those of cv. Labelle. We also observed a gene dosage effect of the Wx(a) transgene on Wx protein expression, but a smaller dosage effect was observed in amylose production with over 40% of amylose content in transgenic rice. Moreover, one transgenic line carrying eleven copies of the transgene showed low levels of Wx expression and amylose in the endosperm. This suggested that the integration of excessive copies of the transgene might lead to gene silencing. PMID- 12773632 TI - Characterization of a rice chlorophyll-deficient mutant using the T-DNA gene-trap system. AB - We have previously generated a large pool of T-DNA insertional lines in rice. In this study, we screened those T-DNA pools for rice mutants that had defective chlorophylls. Among the 1,995 lines examined in the T2 generation, 189 showed a chlorophyll-deficient phenotype that segregated as a single recessive locus. Among the mutants, 10 lines were beta-glucuronidase (GUS)-positive in the leaves. Line 9-07117 has a T-DNA insertion into the gene that is highly homologous to XANTHA-F in barley and CHLH in Arabidopsis: This OsCHLH gene encodes the largest subunit of the rice Mg-chelatase, a key enzyme in the chlorophyll branch of the tetrapyrrole biosynthetic pathway. In the T2 and T3 generations, the chlorina mutant phenotypes are co-segregated with the T-DNA. We have identified two additional chlorina mutants that have a Tos17 insertion in the OsCHLH gene. Those phenotypes were co-segregated with Tos17 in the progeny. GUS assays and RNA blot analysis showed that expression of the OsCHLH gene is light inducible, while TEM analysis revealed that the thylakoid membrane of the mutant chloroplasts is underdeveloped. The chlorophyll content was very low in the OschlH mutants. This is the first report that T-DNA insertional mutagenesis can be used for functional analysis of rice genes. PMID- 12773634 TI - Modulation of pumpkin glutathione S-transferases by aldehydes and related compounds. AB - Induction of pumpkin (Cucurbita maxima Duch.) glutathione S-transferase (GST, EC 2.5.1.18) by aldehydes and related compounds was examined. All of the tested compounds induced pumpkin GST to different degrees, and it was found that (1) aldehydes induce GST directly and alcohols induce GST indirectly, and (2) alpha,beta-unsaturated aldehydes are the most effective inducers and their potency is related to the Michael acceptors reaction. The results of Western blot analysis showed that the patterns of induction of CmGSTU1, CmGSTU2 and CmGSTU3 were similar to the patterns of activity with the exception of alpha,beta unsaturated carbonyl compounds. Among the three compounds, crotonaldehyde caused the highest activity induction (9.2-fold), but Western blot expression was the highest only for CmGSTU3. CmGSTF1 was almost non-responsive to all of the tested stresses. Results of induction studies suggested that efficient pumpkin GST inducers have distinctive chemical features. The in vitro activity of the enzyme was inhibited by ethacryanic acid, trans-2-hexenal, crotonaldehyde, and pentanal. Ethacryanic acid was found to be the most potent inhibitor with an apparent I(50) value of 6.90+/-2.06 micro M, while others were weak to moderate inhibitors. The results presented here indicate that plant GSTs might be involved in the detoxification of physiologically and environmentally hazardous aldehydes/alcohols. PMID- 12773635 TI - A mechanism for light-induced translation of the rbcL mRNA encoding the large subunit of ribulose-1,5-bisphosphate carboxylase in barley chloroplasts. AB - Translational regulation plays a key role in light-induced expression of photosynthesis-related genes at various levels in chloroplasts. We here present the results suggesting a mechanism for light-induced translation of the rbcL mRNA encoding the large subunit (LS) of ribulose-1,5-bisphosphate carboxylase (Rubisco). When 8-day-old dark-grown barley seedlings that have low plastid translation activity were illuminated for 16 h, a dramatic increase in synthesis of large subunit of Rubisco and global activation of plastid protein synthesis occurred. While an increase in polysome-associated rbcL mRNA was observed upon illumination for 16 h, the abundance of translation initiation complexes bound to rbcL mRNA remained constant, indicating that translation elongation might be controlled during this dark-to-light transition. Toeprinting of soluble rbcL polysomes after in organello plastid translation showed that ribosomes of rbcL translation initiation complexes could read-out into elongating ribosomes in illuminated plastids whereas in dark-grown plastids, read-out of ribosomes of translation initiation complexes was inhibited. Moreover, new rounds of translation initiation could also occur in illuminated plastids, but not in dark grown plastids. These results suggest that translation initiation complexes for rbcL are normally formed in the dark, but the transition step of translation initiation complexes entering the elongation phase of protein synthesis and/or the elongation step might be inhibited, and this inhibition seems to be released upon illumination. The release of such a translational block upon illumination may contribute to light-activated translation of the rbcL mRNA. PMID- 12773637 TI - Metabolic flux analysis of the phenylpropanoid pathway in wound-healing potato tuber tissue using stable isotope-labeled tracer and LC-MS spectroscopy. AB - The metabolic flux of two phenylpropanoid metabolites, N-p-coumaroyloctopamine (p CO) and chlorogenic acid (CGA), in the wound-healing potato tuber tissue was quantitatively analyzed by a newly developed method based upon the tracer experiment using stable isotope-labeled compounds and LC-MS. Tuber disks were treated with aqueous solution of L-phenyl-d(5)-alanine, and the change in the ratio of stable isotope-labeled compound to non-labeled (isotope abundance) was monitored for p-CO and CGA in the tissue extract by LC-MS. The time-dependent change in the isotope abundance of each metabolite was fitted to an equation that was derived from the formation and conversion kinetics of each compound. Good correlations were obtained between the observed and calculated isotope abundances for both p-CO and CGA. The rates of p-CO formation and conversion (i.e. fluxes) were 1.15 and 0.96 nmol (g FW)(-1) h(-1), respectively, and for CGA, the rates 4.63 and 0.42 nmol (g FW)(-1) h(-1), respectively. This analysis enabled a direct comparison of the biosynthetic activity between these two compounds. PMID- 12773636 TI - Sucrose synthase catalyzes the de novo production of ADPglucose linked to starch biosynthesis in heterotrophic tissues of plants. AB - By using barley seeds, developmental changes of ADPglucose (ADPG)-producing sucrose synthase (SS) and ADPG pyrophosphorylase (AGPase) have been compared with those of UDPglucose (UDPG), ADPG, sucrose (Suc) and starch contents. Both ADPG synthesizing SS and AGPase activity patterns were found to correlate well with those of ADPG and starch contents. Remarkably, however, maximal activities of ADPG-synthesizing SS were found to be several fold higher than those of AGPase throughout seed development, the highest rate of starch accumulation being well accounted for by SS. Kinetic analyses of SS from barley endosperms and potato tubers in the Suc cleavage direction showed similar K(m) values for ADP and UDP, whereas apparent affinity for Suc was shown to be higher in the presence of UDP than with ADP. Moreover, measurements of transglucosylation activities in starch granules incubated with purified SS, ADP and [U-(14)C]Suc revealed a low inhibitory effect of UDP. The ADPG and UDPG contents in the transgenic S-112 SS and starch deficient potato mutant [Zrenner et al. (1995) Plant J. 7: 97] were found to be 35% and 30% of those measured in wild-type plants, whereas both glucose-1-phosphate and glucose-6-phosphate contents were found to be normal as compared with those of wild-type plants. The overall results thus strongly support a novel gluconeogenic mechanism reported previously [Pozueta-Romero et al. (1999) CRIT: Rev. Plant Sci. 18: 489] wherein SS catalyses directly the de novo production of ADPG linked to starch biosynthesis in heterotrophic tissues of plants. PMID- 12773638 TI - Is Ca2+ release from internal stores involved in membrane excitation in characean cells? AB - An action potential in characean cells is accompanied by an increase in the cytosolic Ca(2+) concentration ([Ca(2+)](c)) which subsequently causes cessation of cytoplasmic streaming. Two Ca(2+ )origins are postulated for the increase in [Ca(2+)](c), extracellular and intracellular ones. For the extracellular origin, a Ca(2+) influx through voltage-dependent Ca(2+)-permeable channels is postulated. For the intracellular origin, a chain of reactions is assumed to occur, involving phosphoinositide-specific phospholipase C (PI-PLC) activation, production of inositol 1,4,5-trisphosphate (IP(3)) and IP(3)-dependent Ca(2+) release from internal stores [Biskup et al. (1999) FEBS Lett. 453: 72]. The hypothesis of the intracellular Ca(2+) origin was tested in three ways: injection of IP(3) into the streaming endoplasm, application of inhibitors of PI-PLC (U73122 and neomycin) and application of an inhibitor of IP(3)-receptor (2 aminoethoxydiphenyl borate; 2APB). Injection of 1 mM IP(3) into Chara cells did not change the rate of cytoplasmic streaming. Both U73122 (20 micro M) and neomycin (200 micro M) did not affect the generation of the action potential, cessation of cytoplasmic streaming and the increase in [Ca(2+)](c) caused by electric stimulus even 20-30 min after application. 2APB depolarized the membrane and inhibited the excitability of the plasma membrane. The results are not consistent with the data obtained by Biskup et al. (1999) who found inhibition of the excitatory inward current by neomycin and U73122. The hypotheses of internal and external Ca(2+) origins are discussed in the light of the present results. PMID- 12773639 TI - Low-temperature induced transmembrane potential changes in the liverwort Conocephalum conicum. AB - Intracellular microelectrode measurements revealed that the liverwort Conocephalum conicum generates all-or-none action potentials (APs) in response to a sudden temperature drop. In plants with anion and potassium conductance blocked, dose-dependent voltage transients (VTs) were evoked by cold stimuli. These VTs did not propagate. When the external concentration of Ca(2+) was decreased or calcium channel inhibitors (La(3+), Gd(3+), verapamil, Mg(2+), Mn(2+)) were used, inhibition of VTs was observed. Amplitudes of both APs and VTs grew when Sr(2+) ions, known to release calcium from internal stores, were added to the medium. Neomycin, which suppresses phospholipase C and indirectly affects inositol triphosphate formation, caused substantial inhibition of both APs and VTs. It is concluded that a temperature drop elucidated membrane potential changes due to calcium influx both from external and internal stores. PMID- 12773640 TI - Effects of low CO2 on NAD(P)H dehydrogenase, a mediator of cyclic electron transport around photosystem I in the cyanobacterium synechocystis PCC6803. AB - The expression and activity of type-1 NAD(P)H dehydrogenase (NDH-1) was compared between cells of Synechocystis PCC6803 grown in high (H-cells) and low (L-cells) CO(2) conditions. Western analysis indicated that L-cells contain higher amounts of the NDH-1 subunits, NdhH, NdhI and NdhK. An NADPH-specific subcomplex of NDH-1 showed higher NADPH-nitroblue tetrazolium oxidoreductase activity in L-cells. The activities of both NADPH-menadione oxidoreductase and light-dependent NADPH oxidation driven by photosystem I were much higher in L-cells than in H-cells. The initial rate of re-reduction of P700(+) following actinic light illumination in the presence of DCMU under background far-red light was enhanced in L-cells. In addition, rotenone, a specific inhibitor of NDH-1, suppressed the relative rate of post-illumination increase in Chl fluorescence of L-cells more than that of H-cells, suggesting that the involvement of NDH-1 in cyclic electron flow around photosystem I was enhanced by low CO(2). Taken together, these results suggest that NDH-1 complex and NDH-1-mediated cyclic electron transport are stimulated by low CO(2) and function in the acclimation of cyanobacteria to low CO(2). PMID- 12773641 TI - Toxicity of free proline revealed in an arabidopsis T-DNA-tagged mutant deficient in proline dehydrogenase. AB - The toxicity of proline (Pro) to plant growth has raised questions despite its protective functions in response to environmental stresses. To evaluate Pro toxicity, we isolated an Arabidopsis T-DNA-tagged mutant, pdh, that had a defect in Pro dehydrogenase (AtProDH), which catalyzes the first step of Pro catabolism. The pdh mutant showed hypersensitivity to exogenous application of < or =10 mM L Pro, at which wild-type plants grew normally. A dose-dependent increase in internal free Pro accumulation was observed in pdh plants during external Pro supply. These results do not just prove the toxicity of Pro, but also suggest that AtProDH is the only enzyme acting as a functional ProDH in Arabidopsis: To further analyze the targets of Pro toxicity, we compared the expression of thousands of genes by pdh plants with that by wild-type plants by cDNA microarray analysis. Most genes were unaffected. Here we demonstrate Pro toxicity by using the pdh mutant and discuss a cause-and-effect action between an excess of free Pro and growth inhibition in Arabidopsis. PMID- 12773642 TI - The CaTin1 (Capsicum annuum TMV-induced clone 1) and CaTin1-2 genes are linked head-to-head and share a bidirectional promoter. AB - CaTin1 was expressed relatively early in the TMV-inoculated leaves of hot pepper which is resistant to TMV-P(0) infection. Interestingly, there was another homologous gene (CaTin1-2) located in front of CaTin1 in a head-to-head fashion and they shared a single promoter. The expression profile of the CaTin1-2 was very similar to CaTin1 in all the treatments except the slower induction time compared to CaTin1 upon TMV-P(0) inoculation. The promoter analysis of CaTin1 and CaTin1-2 revealed bidirectionality both in cis-elements and activity. The CaTin1 2 promoter had two TATA-boxes, four GCC-boxes, the root responsive element, and a W1-box. The ethylene-inducible promoter activity depended on GCC-boxes and TMV inducible activity of the CaTin1-2 promoter reached its highest activity when this promoter had a W1-box. PMID- 12773643 TI - HIV infection--a new drug and new costs. PMID- 12773644 TI - West Nile virus--not a passing phenomenon. PMID- 12773645 TI - Efficacy of enfuvirtide in patients infected with drug-resistant HIV-1 in Europe and Australia. AB - BACKGROUND: The T-20 vs. Optimized Regimen Only Study 2 (TORO 2) compared the efficacy and safety of 24 weeks of treatment with the fusion inhibitor enfuvirtide in combination with an optimized background antiretroviral regimen with the efficacy and safety of the optimized background regimen alone. METHODS: The patients had previous treatment with each of the three classes of antiretroviral drugs, documented resistance to each class, or both and a plasma level of human immunodeficiency virus type 1 (HIV-1) RNA of at least 5000 copies per milliliter. They were randomly assigned in a 2:1 ratio to receive either enfuvirtide (90 mg twice daily) plus a background regimen optimized with the aid of resistance testing (enfuvirtide group) or the background regimen alone (control group). RESULTS: Of the 512 patients who underwent randomization, 335 in the enfuvirtide group and 169 in the control group received at least one dose of study medication and had at least one follow-up measurement of plasma HIV-1 RNA. The median base-line plasma HIV-1 RNA level was 5.1 log10 copies per milliliter in both groups. The median CD4+ cell count was 98.0 cells per cubic millimeter in the enfuvirtide group and 101.5 cells per cubic millimeter in the control group. Patients had a median of seven years of previous treatment and had received a median of 12 antiretroviral drugs. The background regimen comprised a mean of four antiretroviral drugs in both groups. At 24 weeks, the least-squares mean change from base line in the plasma viral load (intention-to-treat, last observation carried forward) was a decrease of 1.429 log10 copies per milliliter in the enfuvirtide group and a decrease of 0.648 log10 copies per milliliter in the control group, a difference of 0.781 log10 copies per milliliter (P<0.001). The mean increase in the CD4+ cell count was greater in the enfuvirtide group (65.5 cells per cubic millimeter) than in the control group (38.0 cells per cubic millimeter, P=0.02). CONCLUSIONS: The addition of enfuvirtide to an optimized background regimen provided significant viral suppression and immunologic benefit over a 24-week period in HIV-1-infected patients who had previously received multiple antiretroviral drugs. PMID- 12773646 TI - Transmission of West Nile virus from an organ donor to four transplant recipients. AB - BACKGROUND: In August 2002, fever and mental-status changes developed in recipients of organs from a common donor. Transmission of West Nile virus through organ transplantation was suspected. METHODS: We reviewed medical records, conducted interviews, and collected blood and tissue samples for testing with a variety of assays. Persons who donated blood to the organ donor and associated blood components were identified and tested for West Nile virus. RESULTS: We identified West Nile virus infection in the organ donor and in all four organ recipients. Encephalitis developed in three of the organ recipients, and febrile illness developed in one. Three recipients became seropositive for West Nile virus IgM antibody; the fourth recipient had brain tissue that was positive for West Nile virus by isolation and nucleic acid and antigen assays. Serum specimens obtained from the organ donor before and immediately after blood transfusions showed no evidence of West Nile virus; however, serum and plasma samples obtained at the time of organ recovery were positive on viral nucleic acid testing and viral culture. The organ donor had received blood transfusions from 63 donors. A review of blood donors and follow-up testing identified one donor who had viremia at the time of donation and who became seropositive for West Nile virus IgM antibodies during the next two months. CONCLUSIONS: Our investigation of this cluster documents the transmission of West Nile virus by organ transplantation. Organ recipients receiving immunosuppressive drugs may be at high risk for severe disease after West Nile virus infection. Blood transfusion was the probable source of the West Nile virus viremia in the organ donor. PMID- 12773647 TI - Pernicious anemia with neuropsychiatric dysfunction in a patient with sickle cell anemia treated with folate supplementation. PMID- 12773648 TI - Images in clinical medicine. Neurologic manifestations of vitamin B12 deficiency. PMID- 12773649 TI - Regionalization and the underuse of angiography in the Veterans Affairs Health Care System as compared with a fee-for-service system. AB - BACKGROUND: Policies to concentrate or regionalize invasive procedures at high volume medical centers are under active consideration. Such policies could improve outcomes among those who undergo procedures while increasing their underuse among those who never reach such centers. We compared the underuse of needed angiography after acute myocardial infarction in a traditional Medicare fee-for-service system with underuse in the regionalized Department of Veterans Affairs (VA) health care system. METHODS: We studied 1665 veterans from 81 VA hospitals and 19,305 Medicare patients from 1530 non-VA hospitals, all of whom were elderly men. We compared adjusted angiography use and one-year mortality among patients for whom angiography was rated as clinically needed. We compared underuse in models before and after controlling for the on-site availability of cardiac procedures. RESULTS: After adjustment for the need for angiography, underuse was present in both groups, but VA patients remained significantly less likely than Medicare patients to undergo angiography (43.9 percent vs. 51.0 percent; odds ratio, 0.75; 95 percent confidence interval, 0.57 to 0.96). After also controlling for on-site availability of cardiac procedures at the admitting hospital, we found no significant difference in the underuse of angiography among VA patients as compared with Medicare patients (odds ratio, 1.02; 95 percent confidence interval, 0.82 to 1.26) or in one-year mortality (odds ratio, 1.08; 95 percent confidence interval, 0.89 to 1.28). CONCLUSIONS: There is underuse of needed angiography after acute myocardial infarction in both the VA and Medicare systems, but the rate of underuse is significantly higher in the VA. These differences appear to be associated with limited on-site availability of cardiac procedures in the regionalized VA health care system. Further work should focus on how regionalization policies could be improved with effective referral and triage processes. PMID- 12773650 TI - Effect of the transformation of the Veterans Affairs Health Care System on the quality of care. AB - BACKGROUND: In the mid-1990s, the Department of Veterans Affairs (VA) health care system initiated a systemwide reengineering to, among other things, improve its quality of care. We sought to determine the subsequent change in the quality of health care and to compare the quality with that of the Medicare fee-for-service program. METHODS: Using data from an ongoing performance-evaluation program in the VA, we evaluated the quality of preventive, acute, and chronic care. We assessed the change in quality-of-care indicators from 1994 (before reengineering) through 2000 and compared the quality of care with that afforded by the Medicare fee-for-service system, using the same indicators of quality. RESULTS: In fiscal year 2000, throughout the VA system, the percentage of patients receiving appropriate care was 90 percent or greater for 9 of 17 quality of-care indicators and exceeded 70 percent for 13 of 17 indicators. There were statistically significant improvements in quality from 1994-1995 through 2000 for all nine indicators that were collected in all years. As compared with the Medicare fee-for-service program, the VA performed significantly better on all 11 similar quality indicators for the period from 1997 through 1999. In 2000, the VA outperformed Medicare on 12 of 13 indicators. CONCLUSIONS: The quality of care in the VA health care system substantially improved after the implementation of a systemwide reengineering and, during the period from 1997 through 2000, was significantly better than that in the Medicare fee-for-service program. These data suggest that the quality-improvement initiatives adopted by the VA in the mid-1990s were effective. PMID- 12773651 TI - Novel therapies based on mechanisms of HIV-1 cell entry. PMID- 12773652 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 17-2003. A 38-year-old woman with fever, headache, and confusion. PMID- 12773653 TI - Fusion inhibition--a major but costly step forward in the treatment of HIV-1. PMID- 12773654 TI - The right care. PMID- 12773655 TI - Case records of the Massachusetts General Hospital--continuing to learn from the patient. PMID- 12773656 TI - Notice of duplicate publication. PMID- 12773657 TI - Inappropriate advertising of dietary supplements. PMID- 12773658 TI - Human growth hormone and aging. PMID- 12773659 TI - Nephropathy induced by contrast medium. PMID- 12773660 TI - Selective estrogen-receptor modulators. PMID- 12773661 TI - Is the match illegal? PMID- 12773662 TI - Readability of informed-consent forms. PMID- 12773663 TI - Residents' suggestions for reducing medical errors. PMID- 12773664 TI - Cardiac rupture. PMID- 12773665 TI - A mutation conferring resistance to imatinib at the time of diagnosis of chronic myelogenous leukemia. PMID- 12773666 TI - Correlation and simple linear regression. AB - In this tutorial article, the concepts of correlation and regression are reviewed and demonstrated. The authors review and compare two correlation coefficients, the Pearson correlation coefficient and the Spearman rho, for measuring linear and nonlinear relationships between two continuous variables. In the case of measuring the linear relationship between a predictor and an outcome variable, simple linear regression analysis is conducted. These statistical concepts are illustrated by using a data set from published literature to assess a computed tomography-guided interventional technique. These statistical methods are important for exploring the relationships between variables and can be applied to many radiologic studies. PMID- 12773667 TI - Understanding and enhancing work performance. PMID- 12773668 TI - Pseudohypocalcemia with MR imaging contrast agents: a cautionary tale. PMID- 12773669 TI - Hard- versus soft-copy interpretation. PMID- 12773670 TI - American College of Radiology Imaging Network: future clinical trials. PMID- 12773671 TI - Gadodiamide administration causes spurious hypocalcemia. AB - PURPOSE: To evaluate the prevalence of spurious hypocalcemia after gadodiamide enhanced magnetic resonance (MR) imaging. MATERIALS AND METHODS: Eight hundred ninety-six inpatients with available serum calcium data obtained before and after gadodiamide-enhanced MR imaging were identified. Changes in serum calcium measurements following gadodiamide administration in 1,049 MR imaging examinations performed in these patients were correlated with gadodiamide dose, renal function, and time between gadodiamide administration and phlebotomy. RESULTS: Following 42 gadodiamide-enhanced examinations, serum calcium measurements spuriously decreased by more than 2 mg/dL (0.5 mmol/L), resulting in laboratory reports of "critical" hypocalcemia (ie, calcium level < 6 mg/dL [1.5 mmol/L]) in 25 examinations. These reduced calcium measurements were correlated with serum creatinine level (r = 0.39, P <.001), gadodiamide dose (r = 0.37, P <.001), and time between gadodiamide injection and phlebotomy (r = -0.28, P <.001). Spurious reductions in calcium measurements after administration of 0.1 mmol of gadodiamide per kilogram of body weight were greater in patients with renal insufficiency (0.6 mg/dL [0.15 mmol/L] +/- 0.5 [0.125, SD]) than in those with normal renal function (0.14 mg/dL [0.035 mmol/L] +/- 0.4 [0.1]) (P <.001). After administration of more than 0.2 mmol/kg of gadodiamide, spurious calcium measurement decreases were greater in patients with renal insufficiency (2.4 mg/dL [0.6 mmol/L] +/- 3.6 [0.9]) than in those with normal renal function (0.4 mg/dL [0.1 mmol/L] +/- 0.7 [0.175]) (P <.001). Patients with renal insufficiency had spuriously low calcium measurements up to 4(1/2) days after gadodiamide administration. Seven patients were inappropriately treated with intravenous calcium and eleven with oral calcium in response to false-positive laboratory reports of critical hypocalcemia. No patient had characteristic symptoms of hypocalcemia or injuries attributed to the inappropriate medical treatment. CONCLUSION: Gadodiamide administration causes spurious hypocalcemia, particularly at doses of 0.2 mmol/kg or higher and in patients with renal insufficiency. PMID- 12773672 TI - Cost-effectiveness targets for multi-detector row CT angiography in the work-up of patients with intermittent claudication. AB - PURPOSE: To determine the costs, sensitivity for detection of significant stenoses, and proportion of equivocal multi-detector row computed tomographic (CT) angiography results in the work-up of patients with intermittent claudication that would make this imaging examination cost-effective compared with gadolinium-enhanced magnetic resonance (MR) angiography. MATERIALS AND METHODS: A decision model was used to compare the societal cost-effectiveness of a new imaging modality with that of gadolinium-enhanced MR angiography. Main outcome measures were quality-adjusted life years (QALYs) and lifetime costs. By using threshold analysis of a given willingness to pay per QALY, target values for costs, sensitivity for detection of significant stenoses, and proportion of cases requiring additional work-up with intraarterial digital subtraction angiography owing to equivocal results of the new modality were determined. The base case evaluated was that of 60-year-old men with severe intermittent claudication and assumed an incremental cost-effectiveness threshold of 100,000 US dollars per QALY. RESULTS: If treatment were limited to angioplasty, a new imaging modality would be cost-effective if the costs were 300 US dollars and the sensitivity was 85%, even if up to 35% of patients needed additional work-up. When both angioplasty and bypass surgery were considered as treatment options, a new imaging modality was cost-effective if the costs were 300 US dollars, the sensitivity was higher than 94%, and 20% of patients required additional work-up. CONCLUSION: Multi-detector row CT angiography, as compared with currently used imaging modalities such as MR angiography, has the potential to be cost-effective in the evaluation of patients with intermittent claudication. PMID- 12773673 TI - Use of abciximab during infrainguinal peripheral vascular interventions: initial experience. AB - PURPOSE: To investigate the use of abciximab as an adjunct during infrainguinal angioplasty. MATERIALS AND METHODS: Abciximab was used in conjunction with recanalization techniques and angioplasty to treat stenoses or occlusions in 16 patients: 10 patients had high-grade stenoses or occlusions longer than 5 cm in the superficial femoral artery and popliteal artery, and six patients had stenoses or occlusions below the popliteal artery. All patients received a bolus of heparin after arterial access was achieved. Abciximab was administered by means of a weight-base nomogram that consisted of an initial bolus followed by an overnight infusion. Technical success was defined as a less than 30% residual stenosis after angioplasty. Patients were carefully followed up for possible postprocedural complications. Platelet counts were monitored for 24 hours. RESULTS: Technical success was achieved in 15 (94%) of 16 patients. One occlusion of the superficial femoral artery could not be crossed. There was a mean postprocedural increase of 0.23 in the ankle-brachial index. There was no significant effect on platelet count. Two minor complications occurred. One major complication occurred; this was a groin hematoma, which required a blood transfusion. CONCLUSION: The use of abciximab as an adjunct during complex infrainguinal arterial interventions appears promising, on the basis of initial experience. PMID- 12773674 TI - Conventional radiography, rapid MR imaging, and conventional MR imaging for low back pain: activity-based costs and reimbursement. AB - PURPOSE: To incorporate personnel and equipment use time in an activity-based cost comparison of conventional radiography and conventional and rapid magnetic resonance (MR) imaging for low back pain (LBP). MATERIALS AND METHODS: At each of four Seattle Lumbar Imaging Project (SLIP) sites, patients were randomized to undergo conventional radiography or rapid MR imaging of the lumbar spine. For sample SLIP patients and for similar non-SLIP patients undergoing conventional lumbar spine MR imaging as usual care in calendar year 2000, measured imaging room use and technologist and radiologist times were multiplied by costs per minute of standard equipment acquisition, personnel compensation, and related expenses. Resulting provider-perspective costs and Seattle area Medicare reimbursements for conventional MR imaging and radiography for calendar year 2001 were used to estimate future "normative" reimbursement for rapid MR imaging. RESULTS: For 23 conventional radiography, 27 rapid MR imaging, and 38 conventional MR imaging examinations timed in calendar year 2000, all rapid MR imaging times exceeded those of conventional radiography but were less than those of conventional MR imaging. All 0.3- and 0.35-T MR imaging room and technologist times exceeded those for 1.5-T MR imaging. Average costs (in 2001 dollars) were $44 for conventional radiography, 126 US dollars for 1.5-T rapid MR imaging, 128 US dollars for 0.3-0.35-T rapid MR imaging, 267 US dollars for 1.5-T conventional MR imaging, and 264 US dollars for 0.3-0.35-T conventional MR imaging. Conclusions regarding cost differences between conventional radiography and rapid MR imaging were robust to plausible parameter value changes evaluated in sensitivity analyses. Conventional radiography reimbursement was 44 US dollars. Applying the ratio of reimbursement (620 US dollars) to costs (264-267 US dollars) for conventional MR imaging to rapid MR imaging costs predicted reimbursement of 292-300 US dollars for the new modality. CONCLUSION: Times and costs for rapid MR imaging are roughly three times those for conventional radiography but about half those for conventional MR imaging for LBP. While current conventional radiography costs exceed reimbursement, current conventional MR and projected rapid MR imaging reimbursements exceed costs. PMID- 12773675 TI - Imaging of osteoid osteoma with dynamic gadolinium-enhanced MR imaging. AB - PURPOSE: To compare dynamic gadolinium-enhanced T1-weighted magnetic resonance (MR) imaging with nonenhanced T1-weighted and T2-weighted MR imaging and thin section computed tomography (CT) for the demonstration of osteoid osteomas. MATERIALS AND METHODS: The images of 11 patients with pathologically proven osteoid osteomas who underwent nonenhanced MR imaging, dynamic gadolinium enhanced MR imaging, and CT were retrospectively reviewed. Images obtained with all three techniques were scored for conspicuity of the osteoid osteoma relative to the surrounding bone. Time-enhancement curves were generated from signal intensity measurements of these lesions and the adjacent bone marrow. The mean imaging scores of the four techniques were compared, and the statistical significance was calculated by using a linear model with terms for method and patient. Pairwise comparisons were made by using the Tukey-Kramer adjustment for multiple comparisons. RESULTS: Compared with CT, dynamic gadolinium-enhanced MR imaging demonstrated the osteoid osteoma equally well in eight of 11 patients and with better conspicuity in three of 11 patients, although this difference was not statistically significant (P =.69). The dynamic gadolinium-enhanced MR images demonstrated the osteoid osteomas significantly better than the nonenhanced T1 weighted (P <.001) and T2-weighted (P <.001) MR images. On the dynamic gadolinium enhanced MR images, nine (82%) of 11 patients had peak enhancement of the osteoid osteoma in the arterial phase with early partial washout, compared with slower, progressive enhancement of the adjacent marrow. This resulted in greatest lesion to marrow contrast material enhancement in the arterial phase. One osteoid osteoma had peak enhancement in the venous phase, and one showed progressive enhancement through all phases to 150 seconds. CONCLUSION: Osteoid osteomas can be imaged with greater conspicuity by using dynamic gadolinium-enhanced instead of nonenhanced MR imaging and with conspicuity equal to or better than that obtained with thin-section CT. PMID- 12773676 TI - Internal derangement of the wrist: indirect MR arthrography versus unenhanced MR imaging. AB - PURPOSE: To compare indirect magnetic resonance (MR) arthrography with unenhanced MR imaging of the wrist for evaluation of the central disk of the triangular fibrocartilage complex (TFCC) and the scapholunate and lunotriquetral interosseous ligaments. MATERIALS AND METHODS: Eighty-six wrists were evaluated at MR imaging (41 indirect MR arthrography and 45 unenhanced MR imaging examinations). Three musculoskeletal radiologists independently evaluated the central disk of the TFCC and scapholunate and lunotriquetral ligaments and compared the results with those of wrist arthroscopy. Sensitivity and specificity were calculated for each of the readers, and the means were obtained. Sensitivities and specificities were compared with the Student t test. RESULTS: Thirty-three tears of the central disk of the TFCC and 13 scapholunate and 18 lunotriquetral ligament tears were identified at arthroscopy. Sensitivities and specificities were 54%-73% and 83%-91%, respectively, in the evaluation of the central disk of the TFCC, with no significant difference between indirect MR arthrography (P =.666) and unenhanced MR imaging (P =.559). Sensitivities and specificities in the evaluation of the scapholunate ligament were 38%-69% and 75% 99%, respectively, with a significant improvement in sensitivity at indirect MR arthrography (P =.017) and no significant difference in specificity (P =.876). Sensitivities in the evaluation of the lunotriquetral ligament were poor, 0%-22%, though the specificities were 88%-99%, with no significant difference between indirect MR arthrography and unenhanced MR imaging (P =.592 and P =.354, respectively, for sensitivity and specificity. CONCLUSION: Indirect MR arthrography significantly improves sensitivity in the evaluation of the scapholunate ligament when compared with unenhanced MR imaging of the wrist but does not significantly improve the ability to evaluate the central disk of the TFCC or the lunotriquetral ligament. PMID- 12773677 TI - The optic nerve tram-track sign. PMID- 12773678 TI - Endovascular treatment of ruptured intracranial aneurysms with detachable coils: long-term clinical and serial angiographic results. AB - PURPOSE: To evaluate the stability of aneurysm occlusion over time, the need for additional treatments, and the long-term clinical outcome of patients, with emphasis on late recurrences of bleeding. MATERIALS AND METHODS: The records of 160 patients with aneurysmal subarachnoid hemorrhage who were treated with coils were retrospectively reviewed. Follow-up angiography was performed 6 and 18 months after coil placement, and the results were classified as complete, near complete, and incomplete occlusion. RESULTS: Six (4%) of the 160 patients experienced procedural mortality or dependency. After a mean follow-up of 36 months, 134 (84%) patients had a good outcome. Outcome was independent of aneurysm size and location and timing of treatment. Reopening of the aneurysm occurred exclusively during the first 6 months after coil placement, mainly in aneurysms larger than 15 mm. Between 6 and 18 months, no change in aneurysm occlusion was observed. Additional coil placement was performed in 15 (9%) patients. After this second coil placement, nine (7%) aneurysms were still incompletely occluded. Additional therapy was performed in eight (5%) patients. Two recurrences of bleeding were observed in two incompletely occluded large aneurysms. No recurrences of bleeding occurred in patients with completely or near completely occluded aneurysms. CONCLUSION: Coil placement is an effective and safe treatment strategy for patients with aneurysmal subarachnoid hemorrhage. If aneurysm occlusion is sufficient at 6 months, the yield of further follow-up angiography is very low. PMID- 12773679 TI - Surgically staged focal liver lesions: accuracy and reproducibility of dual-phase helical CT for detection and characterization. AB - PURPOSE: To assess the accuracy and reproducibility of dual-phase helical computed tomography (CT) in enabling preoperative detection and characterization of surgically staged focal liver lesions. MATERIALS AND METHODS: Surgically and histopathologically proven liver lesions were evaluated by three experienced CT readers. These lesions were present in 77 patients who underwent dual-phase helical CT. Images were interpreted separately by the three blinded reviewers. Each lesion was graded on a nine-point scale of confidence, with 1 being definitely benign, 9 being definitely malignant, and 5 being indeterminate. The chi2 test was used to determine if the distribution of lesion classifications was different between readers. RESULTS: There was a total of 237 lesions: 73 were benign and 164 were malignant. Sensitivity for lesion detection was 69%, 70%, and 71% for the three reviewers, respectively. Specificity was 91%, 86%, and 90%, and the area under the curve for the alternative-free response receiver operating characteristic curve was 0.84, 0.83, and 0.85, respectively. The difference in the distributions of lesion classification between the three reviewers was not statistically significant (P =.67) as determined by chi2 analysis. CONCLUSION: Dual-phase CT has sensitivity of 69%-71% and high specificity (86%-91%) in enabling the detection and characterization of focal liver lesions. Interpretation is highly reproducible, as there is minimal variation between experienced reviewers. PMID- 12773680 TI - Metastasis to regional lymph nodes in patients with esophageal squamous cell carcinoma: CT versus FDG PET for presurgical detection prospective study. AB - PURPOSE: To prospectively compare the accuracy of fluorine 18 fluorodeoxyglucose (FDG) positron emission tomography (PET) and computed tomography (CT) for detection of primary tumor and metastasis to individual lymph node groups and for nodal staging. MATERIALS AND METHODS: From February 2000 to July 2001, 81 patients with squamous cell carcinoma of the esophagus (78 men and three women; age range, 31-90 years; mean age, 63 years) underwent CT and FDG PET before esophagectomy and lymph node dissection. During surgery, all visible and palpable lymph nodes in the surgical fields were removed. The accuracies of CT and FDG PET for depiction of metastasis to lymph nodes were compared. RESULTS: For depiction of malignant nodal groups in each lymph node group, the sensitivity, specificity, and accuracy, respectively, of CT were 11% (11 of 96 nodal groups), 95% (553 of 581), and 83% (564 of 677), whereas those of FDG PET were 30% (29 of 96), 90% (525 of 581), and 82% (554 of 677) (P values: < .001, .009, and .382, respectively). Twenty-eight false-positive interpretations were rendered at CT in evaluations of 11 mediastinal, four hilar, and 13 abdominal nodal groups, and 56 false-positive interpretations were rendered at FDG PET in evaluations of 23 mediastinal, 32 hilar, and one abdominal nodal group. CONCLUSION: FDG PET is more sensitive than CT for depicting nodal metastases in patients with squamous cell carcinoma of the esophagus. FDG PET is slightly less specific than CT for depicting metastases, but the difference in specificity between the two modalities is statistically significant. Both FDG PET and CT have low sensitivity for depicting nodal metastasis. The relatively low specificity of FDG PET for depiction of nodal metastasis compared with that of CT is caused mainly by a high rate of false-positive hilar node interpretations. PMID- 12773682 TI - Case 59: Angiolipoma of the breast. PMID- 12773683 TI - Effects of nonionic intravenous contrast agents at PET/CT imaging: phantom and canine studies. AB - PURPOSE: To investigate the effects of intravenous contrast agents on quantitative values obtained with a combined positron emission tomographic (PET) and computed tomographic (CT) scanner by using several phantoms and a dog. MATERIALS AND METHODS: Fluorine 18 fluorodeoxyglucose (FDG) was mixed with different concentrations of contrast agent with the same syringe (phantom 1), and the phantom was scanned. After image reconstruction with various attenuation maps, radioactivity concentrations were compared. Then, FDG solutions with (phantom 2) or surrounded by (phantom 3) various concentrations of contrast agent were scanned repeatedly, and radioactivity concentration was compared. Finally, PET and CT with and without contrast agent were performed in a dog. PET images were reconstructed by using different attenuation maps, and radioactivity concentrations were compared. The radioactivity concentration on germanium 68 (68Ge)-based corrected images was regarded as standard, and percentage bias, defined as difference divided by measured activity of 68Ge-based corrected images, was assessed. The relationship between the concentration of contrast agent and the percentage bias was assessed with the Pearson coefficient r, and the significance of correlations was evaluated with the Fisher z test. RESULTS: All phantom studies demonstrated that presence of a contrast agent resulted in overestimation of emission data. CT numbers showed a strong positive correlation with the percentage bias in phantoms 2 (r = 0.999) and 3 (r = 0.987); the maximum percentage bias at 1,360 HU reached approximately 45%. These effects were independent of FDG concentration. In a canine model, presence of a contrast agent also increased emission activity, but the percentage bias was less than 15% in the liver and smaller in all other organs except the kidney (26%). CONCLUSION: High concentrations of a contrast agent caused considerable overestimation of apparent tracer activity in phantom studies; however, the emission bias was relatively modest in vivo, except in areas with very high contrast agent concentrations. PMID- 12773684 TI - Normal and ischemic epiphysis of the femur: diffusion MR imaging study in piglets. AB - PURPOSE: To evaluate normal diffusion characteristics in the femur in piglets and changes in diffusion with increasing duration of femoral head ischemia. MATERIALS AND METHODS: Normal epiphyses, physes, and metaphyses of piglets were evaluated with line-scan diffusion imaging (n = 12) and diffusion-tensor imaging (n = 4). Apparent diffusion coefficient (ADC) differences between normal proximal and distal femoral structures, epiphyseal and physeal cartilage, and epiphyseal and metaphyseal marrow were compared (Mann-Whitney test). Short-term femoral ischemia was investigated after maximal abduction of the hips for 3 hours (n = 6); ADCs before and after abduction were compared (Wilcoxon signed rank test). Prolonged ischemia was investigated with placement of a ligature around the neck of a femur (n = 7); the ADC of the femur in this condition was compared (Wilcoxon signed rank test) with that of the normal contralateral femur. Changes in ADC ratios at three durations of ischemia (Kruskal-Wallis test) were compared. RESULTS: ADC was greater in epiphyseal cartilage (mean +/- 1 SD, 1.62 x 10(-3) mm2/sec +/- 0.38) than it was in physeal cartilage (1.28 x 10(-3) mm2/sec +/- 0.31) (P <.007) and greater in epiphyseal marrow (1.26 x 10(-3) mm2/sec +/- 0.38) than it was in metaphyseal marrow (0.91 x 10(-3) mm2/sec +/- 0.35) (P <.001). There was columnar arrangement of tensors in the physis. ADC decreased 26% after 3 hours of maximal abduction. After femoral neck ligature, ADC increased a mean of 27% after 6 hours and a mean of 75% after 96 hours. CONCLUSION: Normal line-scan diffusion imaging findings indicate relative restriction of diffusion in the metaphysis and parallel orientation of tensors in the physis. Diffusion is initially restricted with decreased blood flow but increases if ischemia lasts longer. PMID- 12773685 TI - Breast lesions detected with MR imaging: utility and histopathologic importance of identification with US. AB - PURPOSE: To determine the utility and histopathologic importance of ultrasonographic (US) depiction of breast lesions detected with magnetic resonance (MR) imaging. MATERIALS AND METHODS: Retrospective review was performed of 654 consecutive breast MR examinations performed from November 1999 to February 2001. This yielded 64 patients with 93 suspicious, nonpalpable, mammographically occult lesions evident on MR images and recommended for biopsy, for which directed US assessment was performed. Images, pathologic records, and medical records of these patients were reviewed for imaging findings, patient demographics, and histopathologic findings from subsequent biopsy. Statistical analysis was performed with the Fisher exact test. RESULTS: The median size of MR abnormalities was 0.9 cm (range, 0.3-5.0 cm). The pattern of enhancement was a focal mass in 76 (82%) lesions and nonmass in 17 (18%). A US correlate was identified in 21 (23%) lesions detected with MR--19 (25%) of 76 focal mass and two (11%) of 17 nonmass lesions. Carcinoma was found in nine (43%) of these lesions, of which seven (78%) were invasive carcinoma and two (22%) were ductal carcinoma in situ (DCIS). Among the lesions without a US correlate, 10 (14%) yielded carcinoma, of which five (50%) were invasive carcinoma and five (50%) were DCIS. The frequency of cancer was significantly higher for lesions that were detected with MR imaging and had a US correlate than for those that did not have a US correlate (43% vs 14%, P =.01). Overall, 19 (20%) lesions detected with MR imaging and examined with US yielded carcinoma at biopsy, and nine (47%) malignant lesions were seen on US images. US depicted seven (58%) of 12 invasive cancers and two (29%) of seven instances of DCIS. CONCLUSION: The likelihood of carcinoma was significantly higher among lesions with a US correlate (43% carcinoma) than lesions without a US correlate (14% carcinoma). PMID- 12773686 TI - Quantitative assessment of left ventricular function with interactive real-time spiral and radial MR imaging. AB - An interactive real-time spiral gradient-echo and an interactive real-time radial steady-state free precession sequence were investigated for the quantitative assessment of left ventricular function. Data were acquired in 18 patients without electrocardiographic triggering and breath holding. With the interactive real-time spiral gradient-echo sequence, significant underestimation of endocardial and epicardial volumes was demonstrated; with the interactive real time radial steady-state free precession sequence, excellent agreement was shown with standard cardiac-triggered segmented k-space breath-hold steady-state free precession MR imaging. Interactive real-time radial steady-state free precession imaging allows accurate quantitative assessment of left ventricular volumes. PMID- 12773687 TI - Three-dimensional CT angiography of the hepatic artery: use of multi-detector row helical CT and a contrast agent. AB - The administration of a contrast agent to obtain optimal images at three dimensional computed tomographic (CT) angiography of the hepatic artery by using multi-detector row helical CT was investigated. Three-dimensional CT angiographic images were evaluated visually, and quantitative evaluation was performed by measuring the postcontrast CT number of the aorta. The injection rate of 5 mL/sec was significantly superior to that of 4 mL/sec in both evaluations. At administration of a contrast agent with 300 or 350 milligrams of iodine per milliliter, there were no significant differences in either evaluations. The preferable injection rate to obtain sufficient three-dimensional CT angiographic data was 5 mL/sec. PMID- 12773688 TI - Metallic renal artery MR imaging stent: artifact-free lumen visualization with projection and standard renal MR angiography. AB - A cardiac-triggered free-breathing three-dimensional (3D) balanced fast field echo projection renal magnetic resonance (MR) angiographic sequence was investigated for in-stent lumen visualization of a dedicated metallic renal artery stent. Fourteen prototype stents were deployed in the renal arteries of six pigs (in two pigs, three stents were deployed). Projection renal MR angiography was compared with standard contrast material-enhanced 3D breath-hold MR angiography. Artifact-free in-stent lumen visualization was achieved with both projection MR angiography and contrast-enhanced MR angiography. These promising results warrant further studies for visualization of in-stent restenosis. PMID- 12773689 TI - Screening mammography. PMID- 12773690 TI - MR fluid sign in osteoporotic vertebral fracture. PMID- 12773691 TI - Pneumothorax detection with thoracic US: more research is needed. PMID- 12773692 TI - Hemodialysis arteriovenous fistula maturity: US evaluation. PMID- 12773693 TI - Role of MR imaging and iodine 123 MIBG scintigraphy in staging of pediatric neuroblastoma. PMID- 12773694 TI - Intestinal pathophysiology in autism. AB - Autism is a life-long developmental disorder affecting as many as 1 in 500 children. The causes for this profound disorder are largely unknown. Recent research has uncovered pathology in the gastrointestinal tract of autistic children. The pathology, reported to extend from the esophagus to the colon, is described here along with other studies pointing to a connection between diet and the severity of symptoms expressed in autism. The evidence that there is impaired intestinal permeability in autism is reviewed, and various theories are discussed by which a leaky gut could develop. Lastly, some possible ways in which impaired gastrointestinal function might influence brain function are discussed. PMID- 12773695 TI - Asbestos-induced pulmonary toxicity: role of DNA damage and apoptosis. AB - Asbestos causes asbestosis and various malignancies by mechanisms that are not clearly defined. Here, we review the accumulating evidence showing that asbestos is directly genotoxic by inducing DNA strand breaks (DNA-SB) and apoptosis in relevant lung target cells. Although the exact mechanisms by which asbestos causes DNA damage and apoptosis are not firmly established, some of the implicated mechanisms include the generation of iron-derived reactive oxygen species (ROS) as well as reactive nitrogen species (RNS), alteration in the mitochondrial function, and activation of the death receptor pathway. We focus on the accumulating evidence implicating ROS. DNA repair mechanisms have a key role in limiting the extent of DNA damage. Recent studies show that asbestos activates DNA repair enzymes such as apurinic/apyrimidinic endonuclease (APE) and poly (ADP ribose) polymerase (PARP). Asbestos-induced neoplastic transformation may result in the setting where DNA damage overwhelms DNA repair in the face of a persistent proliferative signal. Strategies aimed at limiting asbestos-induced oxidative stress may reduce DNA damage and, as such, prevent malignant transformation. PMID- 12773696 TI - Neurodevelopmental disorders after thimerosal-containing vaccines: a brief communication. AB - We were initially highly skeptical that differences in the concentrations of thimerosal in vaccines would have any effect on the incidence rate of neurodevelopmental disorders after childhood immunization. This study presents the first epidemiologic evidence, based upon tens of millions of doses of vaccine administered in the United States, that associates increasing thimerosal from vaccines with neurodevelopmental disorders. Specifically, an analysis of the Vaccine Adverse Events Reporting System (VAERS) database showed statistical increases in the incidence rate of autism (relative risk [RR] = 6.0), mental retardation (RR = 6.1), and speech disorders (RR = 2.2) after thimerosal containing diphtheria, tetanus, and acellular pertussis (DTaP) vaccines in comparison with thimerosal-free DTaP vaccines. The male/female ratio indicated that autism (17) and speech disorders (2.3) were reported more in males than females after thimerosal-containing DTaP vaccines, whereas mental retardation (1.2) was more evenly reported among male and female vaccine recipients. Controls were employed to determine if biases were present in the data, but none were found. It was determined that overall adverse reactions were reported in similar aged populations after thimerosal-containing DTaP (2.4 +/- 3.2 years old) and thimerosal-free DTaP (2.1 +/- 2.8 years old) vaccinations. Acute control adverse reactions such as deaths (RR = 1.0), vasculitis (RR = 1.2), seizures (RR = 1.6), ED visits (RR = 1.4), total adverse reactions (RR = 1.4), and gastroenteritis (RR = 1.1) were reported similarly after thimerosal-containing and thimerosal-free DTaP vaccines. An association between neurodevelopmental disorders and thimerosal containing DTaP vaccines was found, but additional studies should be conducted to confirm and extend this study. PMID- 12773697 TI - Dietary magnesium intake influences circulating pro-inflammatory neuropeptide levels and loss of myocardial tolerance to postischemic stress. AB - Severe dietary Mg restriction (Mg(9), 9% of recommended daily allowance [RDA], plasma Mg = 0.25 mM) induces a pro-inflammatory neurogenic response in rats (substance P [SP]), and the associated increases in oxidative stress in vivo and cardiac susceptibility to ischemia/reperfusion (I/R) injury were previously shown to be attenuated by SP receptor blockade and antioxidant treatment. The present study assessed if less severe dietary Mg restriction modulates the extent of both the neurogenic/oxidative responses in vivo and I/R injury in vitro. Male Sprague Dawley rats maintained on Mg(40) (40% RDA, plasma Mg = 0.6 mM) or Mg(100) (100% RDA, plasma Mg = 0.8 mM) diets were assessed for plasma SP levels (CHEM-ELISA) during the first 3 weeks and were compared with the Mg(9) group; red blood cell (RBC) glutathione and plasma malondialdehyde levels were compared at 3 weeks in Mg(9), Mg(20) (plasma Mg = 0.4 mM), Mg(40), and Mg(100) rats; and 40-min global ischemia/30-min reperfusion hearts from 7-week-old Mg(20), Mg(40), and Mg(100) rats were compared with respect to functional recovery (cardiac work, and diastolic, systolic, and developed pressures), tissue LDH release, and free radical production (ESR spectroscopy and alpha-phenyl-N-tert butylnitrone [PBN; 3 mM] spin trapping). The Mg(40) diet induced smaller elevations in plasma SP (50% lower) compared with Mg(9), but with a nearly identical time course. RBC glutathione and plasma malondialdehyde levels revealed a direct relationship between the severity of oxidative stress and hypomagnesemia. The dominant lipid free radical species detected in all I/R groups was the alkoxyl radical (PBN/alkoxyl: alpha(H) = 1.93 G, alpha(N) = 13.63 G); however, Mg(40) and Mg(20) hearts exhibited 2.7- and 3.9-fold higher alkoxyl levels, 40% and 65% greater LDH release, and lower functional recovery (Mg(20) < Mg(40)) compared with Mg(100). Our data suggest that varying dietary Mg intake directly influences the magnitude of the neurogenic/oxidative responses in vivo and the resultant myocardial tolerance to I/R stress. PMID- 12773698 TI - Chronically administered acetaminophen and the ischemia/reperfused myocardium. AB - Male and female Hartley strain guinea pigs weighing 280 +/- 10 g were given acetaminophen-treated water ad libitum for 10 days. Sham-treated control animals were given similar quantities of untreated tap water (vehicle-treated control group). On Day 10, hearts were extracted, instrumented, and exposed to an ischemia (low-flow, 20 min)/reperfusion protocol. Our objective was to compare and contrast ventricular function, coronary circulation, and selected biochemical and histological indices in the two treatment groups. Left ventricular developed pressure in the early minutes of reperfusion was significantly greater in the presence of acetaminophen, e.g., at 1 min, 40 +/- 4 vs 21 +/- 3 mmHg (P < 0.05). Coronary perfusion pressure was significantly less from 3 to 40 min of reperfusion in the presence of acetaminophen. Creatine kinase release in vehicle treated hearts rose from 42 +/- 14 (baseline) to 78 +/- 25 units/liter by the end of ischemia. Corresponding values in acetaminophen-treated hearts were 36 +/- 8 and 44 +/- 14 units/liter. Acetaminophen significantly (P < 0.05) attenuated release of creatine kinase. Chemiluminescence, an indicator of the in vitro production of peroxynitrite via the in vivo release of superoxide and nitric oxide, was also significantly attenuated by acetaminophen. Electron microscopy indicated a well-preserved myofibrillar ultrastructure in the postischemic myocardium of acetaminophen-treated hearts relative to vehicle-treated hearts (e.g., few signs of contraction bands, little or no evidence of swollen mitochondria, and well-defined light and dark bands in sarcomeres with acetaminophen; opposite with vehicle). We conclude that chronic administration of acetaminophen provides cardioprotection to the postischemic, reperfused rodent myocardium. PMID- 12773699 TI - The effect of fructooligosaccharides with various degrees of polymerization on calcium bioavailability in the growing rat. AB - Maximizing peak bone mass during adolescence may be the key to postponing and perhaps preventing bone fractures due to osteoporosis in later life. One mechanism to maximize peak bone mass is to maximize calcium absorption, and it has been suggested that inulin and oligofructose might be one of the ways of doing so. In this study, fructooligosaccharides with various degrees of polymerization have been compared in terms of impact on calcium absorption, bone density, and excretion of collagen cross-links in the young adult male rat. The various oligosaccharides were oligofructose (DP2-8), inulin (DP>23), and a mixture of 92% inulin and 8% short-chain oligofructose (DP2-8). Measuring ex vivo bone mineral density (BMD) and bone mineral content (BMC) showed that BMD was significantly higher in the group fed inulin (DP>23) in both femurs, whereas BMC was significantly higher in the spine. The excretion of fragments of Type 1 collagen decreased in all groups over the 4 weeks of feeding, but the decrease was most significant in the group fed inulin (DP>23). Several hypotheses have been offered to explain the effect of the fructooligosaccharides on calcium absorption and retention. These include the production of organic acids that would acidify the luminal contents and enhance solubility and hence absorption, or possibly a mechanism via calbindinD9k. This study is unique in that it compares the different fructooligosaccharides in the same model, and it clearly shows that the various fructans do not have the same effect. In our model, inulin (DP>23) had the most significant effect on calcium bioavailability. PMID- 12773700 TI - Zinc accumulation in N-methyl-N-nitrosourea-induced rat mammary tumors is accompanied by an altered expression of ZnT-1 and metallothionein. AB - Zinc is essential for cell proliferation. Several human studies have shown that in breast cancer tissues, zinc concentration expressed on a per tissue weight basis is higher than that in normal breast tissues. However, the mechanisms involved are unknown. N-methyl-N-nitrosourea (MNU)-induced rat mammary tumorigenesis is one of the most widely used rodent mammary tumorigenesis models for studying human breast cancer due to their similarities in hormone dependency, pathogenesis, histological classification, and immunocytochemical markers. This study was to establish if there was an accumulation of zinc in MNU-induced rat mammary tumors and, if there was, to explore the possible mechanisms involved. Sprague-Dawley rats were sham-treated or MNU-treated (50 mg/kg; n = 12) for 100 days. In MNU-induced mammary tumors (mammary tumors), zinc concentration expressed on a per dry weight basis was 12 times of that in normal mammary glands. Moreover, the mRNA level of ZnT-1 (a transporter involved in zinc efflux) in mammary tumors was reduced by 55% as compared with that in normal mammary glands. The mRNA level of Nramp2 (a divalent cation importer) and ZnT-4 (another transporter involved in zinc efflux) was unaffected by MNU-induced mammary tumorigenesis. The mRNA and protein levels of metallothionein (a putative zinc storage protein) in mammary tumors were 1.3 and 3.5 times of that in normal mammary glands, respectively. Collectively, our observations showed that zinc is accumulated in MNU-induced rat mammary tumors and this accumulation is accompanied by an altered expression of ZnT-1 and metallothionein, suggesting that zinc homeostasis might be altered in MNU-induced rat mammary tumorigenesis. Because zinc is essential to cell proliferation and cell proliferation is increased in mammary tumors, zinc accumulation is likely a part of an integrated effort to ensure sufficient zinc supply to sustain tumor growth. PMID- 12773701 TI - Regulation of vascular endothelial growth factor gene expression in murine macrophages by nitric oxide and hypoxia. AB - Vascular endothelial growth factor (VEGF) expression in murine peritoneal macrophages is strongly upregulated by hypoxia via transcriptional and posttranscriptional mechanisms. Interferon-gamma (IFN-gamma) with Escherichia coli lipopolysaccharide (LPS) also upregulates expression of VEGF, as well as of the inducible nitric oxide synthase (iNOS). Hypoxia (1% O(2)) upregulates VEGF expression in macrophages from both wild-type and iNOS knockout mice, indicating that hypoxic upregulation of VEGF is independent of iNOS. However, the iNOS inhibitor aminoguanidine (AG) decreases the VEGF expression induced by LPS/IFN gamma, indicating an important role for NO. NO-dependent induction of VEGF is strongly dependent on cell density. LPS/IFN-gamma treatment induces minimal VEGF protein expression in macrophages cultured at low cell densities (<0.25 x 10(6) cells/cm(2)); at higher cell densities (>0.25 x 10(6) cells/cm(2)) that lead to conditions of pericellular hypoxia, however, induction of VEGF expression was strong. Transient transfection of RAW 264.7 cells with luciferase reporter constructs of the murine VEGF promoter indicates that both hypoxia and LPS/IFN gamma independently induce VEGF promoter activity, irrespective of cell density. Although LPS/IFN-gamma treatment induces transcriptional activation of the VEGF promoter, significant levels of VEGF protein are only expressed by cells at high density under conditions of pericellular hypoxia. This suggests an important regulatory role for hypoxia at the posttranscriptional level. Deletion analysis of the VEGF promoter shows that the hypoxia response element region and its immediate flanking sequences are essential for both hypoxia and LPS/IFN-gamma induced VEGF promoter activation. PMID- 12773702 TI - Myostatin gene expression is reduced in humans with heavy-resistance strength training: a brief communication. AB - This study examined changes in myostatin gene expression in response to strength training (ST). Fifteen young and older men (n = 7) and women (n = 8) completed a 9-week heavy-resistance unilateral knee extension ST program. Muscle biopsies were obtained from the dominant vastus lateralis before and after ST. In addition to myostatin mRNA levels, muscle volume and strength were measured. Total RNA was reverse transcribed into cDNA, and myostatin mRNA was quantified using quantitative PCR by standard fluorescent chemistries and was normalized to 18S rRNA levels. A 37% decrease in myostatin expression was observed in response to ST in all subjects combined (2.70 +/- 0.36 vs 1.69 +/- 0.18 U, arbitrary units; P < 0.05). Though the decline in myostatin expression was similar regardless of age or gender, the small number of subjects in these subgroups suggests that this observation needs to be confirmed. No significant correlations were observed between myostatin expression and any muscle strength or volume measure. Although further work is necessary to clarify the findings, these data demonstrate that myostatin mRNA levels are reduced in response to heavy-resistance ST in humans. PMID- 12773703 TI - Subcutaneous adipose tissue pattern in lean and obese women with polycystic ovary syndrome. AB - The new optical device, Lipometer, permits the noninvasive, quick, safe, and precise measurement of the thickness of subcutaneous adipose tissue (SAT) layers at any given site of the human body. Fifteen anatomically well-defined body sites from neck to calf describe the SAT topography (SAT-Top) like an individual "fingerprint." SAT-Top was examined in 33 women with polycystic ovary syndrome (PCOS), in 87 age-matched healthy controls and in 20 Type-II diabetic women. SAT Top differences of these three groups were described, and, based on a hierarchical cluster analysis, two distinctly different groups of PCOS women, a lean (PCOS(L)) and an obese (PCOS(O)) cluster, were found. For visual comparison of the different types of body fat distribution, the 15-dimensional body fat information was condensed to a two-dimensional factor plot by factor analysis. For comparison of the PCOS like body fat distribution with the "healthy" fat pattern, the (previously published) SAT-Top results of 590 healthy women and men (20-70 years old) and 162 healthy girls and boys (7-11 years old) were added to the factor plot. PCOS(O) women showed a SAT-Top pattern very similar to that of women with Type-II diabetes, even though the diabetic women were on average 30 years older. Compared with their healthy controls, SAT-Top of these PCOS(O) patients was strongly skewed into the android direction, providing significantly decreased leg SAT development and significantly higher upper body obesity. Compared with healthy women, PCOS(L) patients had significantly lower total SAT development (even though height, weight, and body mass index did not deviate significantly), showing a slightly lowered amount of body fat in the upper region and a highly significant leg SAT reduction. This type of fat pattern is the same as found in girls and boys before developing their sex specific body fat distribution. We conclude that women with PCOS develop an android SAT-Top, but compared in more detail, we found two typical types of body fat distribution: the "childlike" SAT pattern in lean PCOS patients, and the "diabetic" body fat distribution in obese PCOS women. PMID- 12773704 TI - Effects of hypoxia on the development of intestinal enzymes in neonatal and juvenile rats. AB - Hypoxia in the neonate is known to alter the activity of hepatic and pancreatic enzymes involved in lipid and carbohydrate metabolism. The purpose of this study was to evaluate the effect of neonatal hypoxia on the activity of intestinal enzymes, and to determine whether the administration of glucocorticoids to neonates can mimic the effects of hypoxia. Hypoxia in neonatal rats (0-7 days) increased protein content, and lactase and maltase activity in the duodenal and the jejunal segments of the small intestine compared with normoxic controls. Hypoxia in juvenile rats (28-35 days) did not change these enzymes. Two weeks after returning hypoxic (0-7 days) pups to normoxia, their body weight remained lower than the age-matched controls. In the group recovering from hypoxia, sucrase, maltase, and leucine aminopeptidase activities were lower in the duodenal and the jejunal segment. Compared with controls, LDH activity was lower only in the jejunal intestine in the group recovering from hypoxia. All enzyme activities returned to control levels 3 weeks after recovery. Neonatal rats treated with dexamethasone had a decrease in body weight, but increases in sucrase and maltase activity in both the duodenal and the jejunal segment. Hypoxia in newborn rats caused a delayed maturation of small intestinal enzymes. Increases in serum glucocorticoids after hypoxic exposure probably do not play a major role in the delayed maturation of the disaccharidase activity in the small intestine. PMID- 12773705 TI - Promotive effects of far-infrared ray on full-thickness skin wound healing in rats. AB - The biological effects of far-infrared ray (FIR) on whole organisms remain poorly understood. The aim of our study was to investigate not only the hyperthermic effect of the FIR irradiation, but also the biological effects of FIR on wound healing. To evaluate the effect of FIR on a skin wound site, the speed of full thickness skin wound healing was compared among groups with and without FIR using a rat model. We measured the skin wound area, skin blood flow, and skin temperature before and during FIR irradiation, and we performed histological inspection. Wound healing was significantly more rapid with than without FIR. Skin blood flow and skin temperature did not change significantly before or during FIR irradiation. Histological findings revealed greater collagen regeneration and infiltration of fibroblasts that expressed transforming growth factor-beta1 (TGF-beta1) in wounds in the FIR group than in the group without FIR. Stimulation of the secretion of TGF-beta1 or the activation of fibroblasts may be considered as a possible mechanisms for the promotive effect of FIR on wound healing independent of skin blood flow and skin temperature. PMID- 12773706 TI - Norepinephrine enhances adhesion of HIV-1-infected leukocytes to cardiac microvascular endothelial cells. AB - Recent reports have indicated that norepinephrine (NE) enhances HIV replication in infected monocytes and promotes increased expression of select matrix metalloproteinases associated with dilated cardiomyopathy (DCM) in vitro in co cultures of HIV-infected leukocytes and human cardiac microvascular endothelial cells (HMVEC-C). The influence of NE on HIV infection and leukocyte-endothelial interactions suggests a pathogenic role in AIDS-related cardiovascular disease. This study examined the effects of norepinephrine (NE) and HIV-1 infection on leukocyte adhesion to HMVEC-C. Both flow and static conditions were examined and the expression of selected adhesion molecules and cytokines were monitored in parallel. NE pretreatment resulted in a detectable, dose-dependent increase of leukocyte-endothelial adhesion (LEA) with both HIV-1-infected and -uninfected peripheral blood mononuclear cells (PBMCs) relative to media controls after 48 hr in co-culture with HMVEC-C in vitro. However, the combination of NE plus HIV infection resulted in a significant (P < 0.0001) 18-fold increase in LEA over uninfected media controls. Increased levels in both cell-associated and -soluble ICAM-1 and E-Selectin but not VCAM-1 correlated with increased LEA and with HIV-1 infection or NE pretreatment. Blocking antibodies specific for ICAM-1 or E Selectin inhibited HIV-NE-induced LEA. These data suggest a model in which NE primes HIV-1-infected leukocytes for enhanced adhesion and localization in HMVEC C where they can initiate and participate in vascular injury associated with AIDS related cardiomyopathy. PMID- 12773707 TI - Influence of nitric oxide on the secretory function of the bovine corpus luteum: dependence on cell composition and cell-to-cell communication. AB - The objective of the present study was to investigate the role of cell-to-cell contact in the influence of nitric oxide (NO) on the secretory function of the bovine corpus luteum (CL). In Experiment 1, separate small luteal cells (SLC) or large (LLC) luteal cells were perfused with 100 micro M spermineNONOate, a NO donor, or with 100 micro M Nomega-nitro-L-arginine methyl ester (L-NAME), a NO synthase (NOS) inhibitor; in Experiment 2, a mixture of LLC and SLC and endothelial cells was cultured and incubated with spermineNONOate or L-NAME; in Experiment 3, spermineNONOate was perfused into the CL (100 mg/4 hr) by a microdialysis system in vivo. Perfusion of isolated SLC and LLC with the NO donor or NOS inhibitor (Experiment 1) did not affect (P > 0.05) secretion of progesterone (P(4)) or oxytocin (OT). L-NAME perfusion increased (P < 0.05) leukotriene C(4) (LTC(4)) secretion by both SLC and LLC cells. Treatment of mixtures of luteal cells with an NO donor (Experiment 2) significantly decreased (P < 0.001) secretion of P(4) and OT and increased (P < 0.001) production of prostaglandin F(2alpha) (PGF(2alpha)) and LTC(4). L-NAME stimulated (P < 0.001) P(4) secretion, but did not influence (P > 0.05) OT, PGF(2alpha) or LTC(4) production. Intraluteal administration (Experiment 3) of spermineNONOate increased (P < 0.001) LTC(4) and PGF(2alpha), decreased OT, but did not change P(4) levels in perfusate samples. These data indicate that cell-to-cell contact and cell composition play important roles in the response of bovine CL to treatment with NO donors or NOS inhibitors, and that paracrine mechanisms are required for the full secretory response of the CL in NO action. Endothelial cells appear to be required for the full secretory response of the CL to NO. PMID- 12773708 TI - Activation of selective transcription factors and cytokines by water-soluble extract from Lentinus lepideus. AB - We isolated a water-soluble extract, PG101, from cultured mycelia of Lentinus lepideus. Treatment of human peripheral blood mononuclear cells (PBMCs) with PG101 increased levels of TNF-alpha, IL-1beta, IL-10, and IL-12 by 100- to 1000 fold, whereas GM-CSF and IL-18 were activated by an order of magnitude. On the contrary, IFN-gamma and IL-4 were not affected. The response to PG101 occurred in a dose- and time-dependent manner. From the human PBMCs treated with PG101, TNF alpha was a first cytokine to be activated, detectable at 2 hr post-treatment followed by IL-1beta at 6 hr post-treatment. IL-12 and IL-10 were the next to follow. GM-CSF and IL-18 both showed significant increases 24 hr after treatment. When PBMCs were sorted into various cell types, monocyte/macrophages, but not T and B cells, were the major target cell type responsive to PG101. Consistent with this result, the profile of cytokine expression upon PG101 treatment was comparable between PBMCs and a human promonocytic cell line (U937), whereas cell lines of T cell and myeloid origins did not respond to PG101. Data from a transient transfection assay involving specific reporter plasmids indicated that cellular transcription factor such as NF-kappaB, but not AP-1, was highly activated by PG101. Results from a gel retardation assay and the experiment involving a specific NF-kappaB inhibitor confirmed the involvement of NF-kappaB. Despite its significant biological effect on various cytokines, PG101 remained nontoxic in both rats and PBMCs even at a biological concentration approximately 20 times greater. PG101 demonstrates great potential as a therapeutic immune modulator. PMID- 12773709 TI - Enhancement of repopulation and hematopoiesis of bone marrow cells in irradiated mice by oral administration of PG101, a water-soluble extract from Lentinus lepideus. AB - PG101 is a water-soluble extract from Lentinus lepideus. It is a potential biological response modifier that activates selective cytokines in vitro, mainly by controlling cellular transcription factor NF-kappaB. Effects of PG101 were tested on bone marrow cells in irradiated mice. Mice were irradiated with a dose of 6 Gy and were given PG101 by gavages daily for 24 days. In PG101-treated mice, the number of colony-forming cells, including colony-forming units (CFU) granulocytes/macrophages (GM) and erythroid burst-forming units (BFU-E), were increased to almost the levels seen in nonirradiated control as early as 8 days after irradiation. Two-color flow cytometric analysis using antibodies to ER-MP12 and ER-MP20 suggested that in the bone marrow cell population, PG101 increased the number of granulocytes (ER-MP12(-)20(med)) and myeloid progenitors (ER MP12(+)20(+)). Analysis of surface c-Kit and Gr-1 proteins in bone marrow cells indicated that PG101 might induce differentiation of progenitor cells to granulocytes and/or proliferation of the committed cells. Lastly, oral administration of PG101 highly increased serum levels of GM-CSF, IL-6, and IL 1beta. Interestingly, the level of TNF-alpha was elevated by irradiation in control mice, but was maintained at the background level in PG101-treated mice, suggesting that PG101 might effectively suppress TNF-alpha-related pathologic conditions. Our results strongly suggest the great potential of PG101 as an immune enhancer during radiotherapy and/or chemotherapy. PMID- 12773710 TI - Other people's smoke: what's in a name? PMID- 12773711 TI - Social sources of cigarettes for youth: broadening the research base. PMID- 12773712 TI - A promise is a promise. AB - Philip Morris promised to "shut down instantly"if cigarettes were found to be harmful. Now that the company has admitted that "smoking causes lung cancer, heart disease, emphysema, and other serious diseases", when will they keep their promise to stop making cigarettes? PMID- 12773713 TI - Vietnam: smuggling adds value. PMID- 12773714 TI - Serbia: tough times ahead. PMID- 12773715 TI - India: PM's bravery awards "nothing to do with our products". PMID- 12773716 TI - Sri Lanka: film's big puff for smoking. PMID- 12773717 TI - USA: Big Tobacco and the lighter side of security. PMID- 12773718 TI - Pakistan: still room for brand launches. PMID- 12773719 TI - Uruguay: ants versus elephants. PMID- 12773720 TI - Estimating the health consequences of replacing cigarettes with nicotine inhalers. AB - BACKGROUND: A fast acting, clean nicotine delivery system might substantially displace cigarettes. Public health consequences would depend on the subsequent prevalence of nicotine use, hazards of delivery systems, and intrinsic hazards of nicotine. METHODS: A spreadsheet program, DEMANDS, estimates differences in expected mortality, adjusted for nicotine delivery system features and prevalence of nicotine use, by extending the data and methods of the SAMMEC 3 software from the US Centers for Disease Control and Prevention. The user estimates disease risks attributable to nicotine, other smoke components, and risk factors that coexist with smoking. The public health consequences of a widely used clean nicotine inhaler replacing cigarettes were compared to historical observations and public health goals, using four different risk attribution scenarios and nicotine use prevalence from 0-100%. MAIN OUTCOME MEASURES: Changes in years of potential life before age 85 (YPL85). RESULTS: If nicotine accounts for less than a third of smokers' excess risk of SAMMEC diseases, as it most likely does, then even with very widespread use of clean nicotine DEMANDS predicts public health gains, relative to current tobacco use. Public health benefits accruing from a widely used clean nicotine inhaler probably equal or exceed the benefits of achieving Healthy People 2010 goals. CONCLUSIONS: Clean nicotine inhalers might improve public health as much as any feasible tobacco control effort. Although the relevant risk estimates are somewhat uncertain, partial nicotine deregulation deserves consideration as part of a broad tobacco control policy. PMID- 12773721 TI - Dimensions underlying legislator support for tobacco control policies. AB - OBJECTIVE: To propose and test a new classification system for characterising legislator support for various tobacco control policies. DESIGN: Cross sectional study. SUBJECTS: Federal and provincial legislators in Canada serving as of October 1996 who participated in the Canadian Legislator Study (n = 553; response rate 54%). MAIN OUTCOME MEASURES: A three factor model (Voters, Tobacco industry, Other interest groups) that assigns nine tobacco control policies according to legislators' hypothesised perceptions of which group is more directly affected by these policies. RESULTS: Based on confirmatory factor analysis, the proposed model had an acceptable fit and showed construct validity. Multivariate analysis indicated that three of the predictors (believing that the government has a role in health promotion, being a non-smoker, and knowledge that there are more tobacco than alcohol caused deaths) were associated with all three factor scales. Several variables were associated with two of the three scales. Some were unique to each scale. CONCLUSIONS: Based on our analyses, legislator support for tobacco control policies can be grouped according to our a priori factor model. The information gained from this work can help advocates understand how legislators think about different types of tobacco control policies. This could lead to the development of more effective advocacy strategies. PMID- 12773723 TI - Development of a model of the tobacco industry's interference with tobacco control programmes. AB - OBJECTIVE: To construct a conceptual model of tobacco industry tactics to undermine tobacco control programmes for the purposes of: (1) developing measures to evaluate industry tactics, (2) improving tobacco control planning, and (3) supplementing current or future frameworks used to classify and analyse tobacco industry documents. DESIGN: Web based concept mapping was conducted, including expert brainstorming, sorting, and rating of statements describing industry tactics. Statistical analyses used multidimensional scaling and cluster analysis. Interpretation of the resulting maps was accomplished by an expert panel during a face-to-face meeting. SUBJECTS: 34 experts, selected because of their previous encounters with industry resistance or because of their research into industry tactics, took part in some or all phases of the project. RESULTS: Maps with eight non-overlapping clusters in two dimensional space were developed, with importance ratings of the statements and clusters. Cluster and quadrant labels were agreed upon by the experts. CONCLUSIONS: The conceptual maps summarise the tactics used by the industry and their relationships to each other, and suggest a possible hierarchy for measures that can be used in statistical modelling of industry tactics and for review of industry documents. Finally, the maps enable hypothesis of a likely progression of industry reactions as public health programmes become more successful, and therefore more threatening to industry profits. PMID- 12773724 TI - Social exchange of cigarettes by youth. AB - OBJECTIVE: This study seeks to determine the magnitude, nature, and correlates of social exchange of cigarettes among youth who smoke. DESIGN: Cross sectional survey. SETTING: Schools in 29 Minnesota communities. PARTICIPANTS: All students in grades 8, 9, and 10 (ages 13-16 years) in each participating school. The parent/child response rate for the survey was 90%. Analyses included 4124 respondents who smoked at least one cigarette in the month before the survey. OUTCOME MEASURES: Social exchange scale, consisting of 16 items assessing aspects of social provision and acquisition of cigarettes. RESULTS: Almost 90% of youth in this study had obtained a cigarette from, and about 75% of them had provided cigarettes to, another teen in the prior month. Daily smokers provided to more teens and provided more often than those who smoked less than daily. Daily smokers also reported having more social sources, both teens and adults, than lighter smokers, and were more likely to have both bought from and sold cigarettes to other teens (p < 0.0001 for all comparisons between daily and less than daily smokers). In a multivariate analysis, social exchange was associated with grade, whether siblings and friends smoke, level of smoking, age of smoking initiation, parental influences and community norms about teen smoking, and buying cigarettes. CONCLUSIONS: Social provision and acquisition of cigarettes among teens are widespread, reciprocal behaviours. Parental and community expectations about smoking influence social exchange, possibly by providing opportunities or barriers for social smoking. Commercial and social availability are not mutually exclusive; rather social exchange extends the reach of commercial sources. PMID- 12773725 TI - Youth preferences for cigar brands: rates of use and characteristics of users. AB - OBJECTIVE: Youth use of cigars has increased in the USA. Understanding brand preference among youth could help explain the attraction to cigars, and develop prevention strategies. This study reports on youth characteristics associated with preferences for cigar brands. DESIGN: Data are presented on 5006 students in grades 7-12 (ages 12-18 years) in classrooms administered the cigar use reasons evaluation (CURE), a questionnaire assessing alternative tobacco use (cigars, bidis, and kreteks). SETTING: Twelve middle and high schools across Massachusetts. MAIN OUTCOME MEASURES: Preferred brands of cigars. RESULTS: Cigar smoking was reported by 16.4% of the sample. The brands preferred by over 5% of cigar users were Phillies (overall prevalence: 31.0%, 95% confidence interval (CI) 27.9% to 34.3%), Black & Mild (12.3%, 95% CI 9.8% to 15.4%), Garcia y Vega (11.4%, 95% CI 9.3% to 13.8%), Swisher Sweets (9.6%, 95% CI 7.6% to 12.2%), Backwoods (6.1%, 95% CI 4.6% to 8.1%), and non-listed brands (16.4%, 95% CI 14.0% to 19.0%). Male cigar smokers were significantly more likely than females to prefer five brands: Backwoods, Dutch Masters, Garcia y Vega, Phillies, and Swisher Sweets, while females were more likely to prefer Black & Mild and non listed brands. Brand preferences were also distinguished by self, friends' and siblings' tobacco use, race/ethnicity, parents' education and cigar use, GPA (grade point average), college plans, and community type. Urban students were more likely to prefer Black & Mild; students whose parents smoke cigars preferred non-listed brands. CONCLUSIONS: Youth prefer certain brands, most notably Phillies. Particular brands are preferred by particular youth subgroups, raising the possibility of selective marketing toward these groups. PMID- 12773726 TI - A method to guide community planning and evaluation efforts in tobacco control using data on smoking during pregnancy. AB - BACKGROUND: Effective community based tobacco control programmes are critical for state and nationwide impact. However, there is little discussion in the literature of methods for setting local objectives which use locally collected data and account for historical variation in progress. OBJECTIVES: To develop and illustrate a method that uses locally available birth certificate data to model trends in tobacco use during pregnancy among women giving birth, predict future prevalence, and use predictions to set community specific tobacco control objectives. DATA SOURCE: Vital statistics. Wisconsin standard birth certificates, 1990-2000, which record the smoking status of the mother during pregnancy. DATA ANALYSIS: Trends in the prevalence of smoking during pregnancy in Wisconsin statewide and in all counties (n = 72) were modelled using linear regression of log prevalence on year. Model fit was assessed using R(2). Regression slopes, indicating estimated relative annual percentage change in prevalence, were used to predict prevalence in 2005, and objectives were calculated as a 20% reduction from the predicted prevalence in 2005. CONCLUSIONS: Modelling trends in the prevalence of smoking using locally collected data enables communities to set reasonable future tobacco control objectives that account for historical trends in progress. PMID- 12773727 TI - A cluster randomised controlled trial of smoking cessation in pregnant women comparing interventions based on the transtheoretical (stages of change) model to standard care. AB - OBJECTIVES: To evaluate the effectiveness in helping pregnant women stop smoking of two interventions (Pro-Change for a healthy pregnancy) based on the transtheoretical model of behaviour change (TTM) compared to current standard care. DESIGN: Cluster randomised trial. SETTING: Antenatal clinics in West Midlands, UK general practices. PARTICIPANTS: 918 pregnant smokers INTERVENTIONS: 100 general practices were randomised into the three trial arms. Midwives in these practices delivered three interventions: A (standard care), B (TTM based self help manuals), and C (TTM based self help manuals plus sessions with an interactive computer program giving individualised smoking cessation advice). MAIN OUTCOME MEASURES: Biochemically confirmed smoking cessation for 10 weeks previously, and point prevalence abstinence, both measured at 30 weeks of pregnancy and 10 days after delivery. RESULTS: There were small differences between the TTM arms. Combining the two arms, the odds ratios at 30 weeks were 2.09 (95% confidence interval (CI) 0.90 to 4.85) for 10 week sustained abstinence and 2.92 (95% CI 1.42 to 6.03) for point prevalence abstinence relative to controls. At 10 days after delivery, the odds ratios were 2.81 (95% CI 1.11 to 7.13) and 1.85 (95% CI 1.00 to 3.41) for 10 week and point prevalence abstinence respectively. CONCLUSIONS: While there is a small borderline significant increase in quitting in the combined intervention arms compared with the controls, the effect of the intervention is small. At 30 weeks gestation and at 10 days postnatal, only about 3% of the intervention groups achieved sustained cessation, with numbers needed to treat of 67 (30 weeks of gestation) and 53 (10 weeks postnatal) for one additional woman to achieve sustained confirmed cessation. Given also that the intervention was resource intensive, it is of doubtful benefit. PMID- 12773728 TI - Improving the self reporting of tobacco use: results of a factorial experiment. AB - OBJECTIVES: To examine divergent estimates of smoking prevalence in two random digit dial surveys for the same population. Based upon internal and external reviews of survey procedures, differences in survey introductions (general health versus tobacco specific introduction) and/or differences in the use of filter questions were identified as the most likely explanations. This prompted an experiment designed to investigate these potential sources of measurement error. DESIGN: A randomised 2 x 2 factorial experiment. SETTING: A random digit dial telephone survey from July to September 2000. SUBJECTS: 3996 adult Californian respondents. MAIN OUTCOME MEASURES: A series of smoking prevalence questions in the context of a tobacco or general health survey. RESULTS: Logistic regression analyses suggest that, among females, prior knowledge (from the survey introduction) that a survey is concerned with tobacco use may decrease self reported smoking prevalence (approximately 4% absolute prevalence difference). Differences in the use of filter questions resulted in almost no misclassification of respondents. CONCLUSIONS: The tobacco specific survey introduction is causing some smokers to deny their tobacco use. The data suggest that these smokers tend to be women that smoked occasionally. A desire by the participants to minimise their personal time costs or a growing social disapproval of tobacco use in the USA may be contributing to the creation of previously undetected survey artefacts in the measurement of tobacco related behaviours. PMID- 12773729 TI - How tobacco companies ensure prime placement of their advertising and products in stores: interviews with retailers about tobacco company incentive programmes. AB - BACKGROUND: About 81% of cigarette manufacturers' marketing expenditures in the USA is spent to promote cigarette sales in stores. Relatively little is known about how these expenditures help the manufacturers achieve their marketing goals in stores. A better understanding of how tobacco companies influence the retail environment would help researchers and tobacco control activists to monitor industry presence in stores. OBJECTIVE: To describe the types of tobacco company incentive programmes offered to retailers, how these programmes impact the store environments, and possible visual indicators of retailer participation in incentive programmes. STUDY DESIGN: In-depth qualitative interviews with a convenience sample of 29 tobacco retailers were conducted in 2001. SETTING: USA. MAIN OUTCOME MEASURES: The types and requirements of retailer incentive programmes provided by tobacco companies, and how participation in a programme alters their stores. RESULTS: The retailers provided insights into how tobacco companies convey promotional allowances and special offers to them and how these incentives shape the retail environment. Retailers noted that tobacco companies exert substantial control over their stores by requiring placement of products in the most visible locations, and of specific amounts and types of advertising in prime locations in the store. Retailers also described how tobacco companies reduce prices by offering them volume based discounts, "buy two, get one free" specials, and "buying down" the price of existing product. CONCLUSIONS: Tobacco companies are concentrating their marketing dollars at the point-of-sale to the extent that the store is their primary communication channel with customers. As a result, all shoppers regardless of age or smoking status are exposed to pro smoking messages. Given the financial resources spent by tobacco companies in stores, this venue warrants closer scrutiny by researchers and tobacco control advocates. PMID- 12773730 TI - Preferences and practices among renters regarding smoking restrictions in apartment buildings. AB - OBJECTIVE: This study assessed renters' preferences for official smoking policies in their buildings and their practices concerning restricting tobacco smoking in their apartments. DESIGN: Renters (n = 301) living in large apartment complexes in a suburb of Minneapolis, Minnesota, completed a mail survey. MAIN OUTCOME MEASURES: The survey asked about the official smoking policies in place in their apartment buildings, their preferences for policies, whether they had smelled tobacco smoke coming into their apartments from without, and, if so, what they had done about it. RESULTS: The majority of non-smokers (79%) preferred that their building be smoke-free. When asked to identify the current smoking policy in their buildings, residents disagreed substantially. Most renters (60%) reported smoke-free policies in their own apartments and another significant proportion (23%) restricted smoking to certain areas or occasions or persons. 75% thought that enforcing a smoke-free policy for guests would not be difficult. 53% of those in non-smoking households had smelled tobacco smoke in their apartments; most of these reported being bothered by it. However, very few complained to the building owner or manager (15.5%) or to the smoker (6.9%). CONCLUSIONS: The majority of non-smokers preferred that their buildings be smoke-free. A failure to report problems to apartment managers might be an impediment to instituting smoke-free policies in apartment buildings. The considerable disagreement among residents within apartment complexes about the current official smoking policy in their buildings suggests that policies are lacking or are not well communicated. PMID- 12773731 TI - Tobacco industry strategies to undermine the 8th World Conference on Tobacco or Health. AB - OBJECTIVE: To demonstrate that Philip Morris and British American Tobacco Company attempted to initiate a wide ranging campaign to undermine the success of the 8th World Conference on Tobacco or Health held in Buenos Aires, Argentina, in 1992. DATA SOURCES: Publicly available tobacco industry documents housed in Minneapolis, Minnesota, USA; Guilford, UK; on-line document websites; and telephone interviews with informed parties. STUDY SELECTION: Those documents determined to be relevant to the companies' campaigns against the 8th World Conference on Tobacco or Health. DATA EXTRACTION: Revision of chapter VIII of the July 2000 WHO report by a committee of experts, entitled: Tobacco company strategies to undermine tobacco control activities at the World Health Organization: report of the committee of experts on tobacco industry documents. DATA SYNTHESIS: Internal documents describe proposed media and science orientated campaigns developed by BAT, Philip Morris, and their consultants to divert attention away from the conference. RESULTS AND CONCLUSION: This work shows that the tobacco industry has the resources and vested interest to combat perceived threats in its regional operating markets, in this case its Latin American market. It is important for the worldwide public heath community to become aware of the numerous ways in which the tobacco industry and its front groups can work against international tobacco control meetings, even including the manipulation of or working with other public health groups to oppose tobacco control efforts. Future world conference planners and participants should be aware that the tobacco industry is likely to continue to employ such methodology. There is no reason to think that the industry is paying less attention to such conferences in the present or future. Rather, it is likely the industry will adopt and expand strategies that were successful while abandoning those that were not effective. Required disclosure of financial support by all participants at all tobacco scientific conferences is recommended. For the tobacco control community, we also recommend careful coalition building and networking with other public health groups on the ways tobacco is implicated in other public health issues. PMID- 12773732 TI - From adversary to target market: the ACT-UP boycott of Philip Morris. AB - BACKGROUND: In 1990, the AIDS Coalition to Unleash Power (ACT-UP) sparked a year long boycott of Philip Morris's Marlboro cigarettes and Miller beer. The boycott protested the company's support of Senator Jesse Helms (R-North Carolina), a leading opponent of AIDS funding and civil rights for lesbian, gay, bisexual and transgender (LGBT) people. ACT-UP demanded that Philip Morris sever its ties with Helms and acknowledge its responsibility to the LGBT community and to people with AIDS. OBJECTIVE: To assess the impact of the boycott on the LGBT community, the tobacco industry, and the tobacco control movement; and to determine what lessons tobacco control advocates can extract from this case. DATA SOURCES: Internal tobacco industry documents and newspaper archives. METHODS: Search of tobacco industry documents websites using "boycott", "ACT-UP", "gay", and other terms. RESULTS: Philip Morris used the boycott to its own advantage. It exploited differences within the community and settled the boycott by pledging large donations to combat AIDS. Through corporate philanthropy, Philip Morris gained entree to the LGBT market without appearing gay friendly. Many LGBT organisations, thirsty for recognition and funding from mainstream corporations, welcomed Philip Morris's overtures without considering the health hazards of tobacco. CONCLUSIONS: Unless the goal of a boycott is to convince the tobacco industry to abandon tobacco altogether, such actions invite the industry to expand its marketing under the guise of philanthropy. Tobacco control advocates should be clear about goals and acceptable settlement terms before participating in a boycott of a tobacco company. PMID- 12773733 TI - Thinking the "unthinkable": why Philip Morris considered quitting. AB - OBJECTIVE: To investigate the genesis and development of tobacco company Philip Morris's recent image enhancement strategies and analyse their significance. DATA SOURCES: Internal Philip Morris documents, made available by the terms of the Master Settlement Agreement between the tobacco companies and the attorneys general of 46 states, and secondary newspaper sources. STUDY SELECTION: Searches of the Philip Morris documents website (www.pmdocs.com) beginning with terms such as "image management" and "identity" and expanding as relevant new terms (consultant names, project names, and dates), were identified, using a "snowball" sampling strategy. FINDINGS AND CONCLUSIONS: In the early 1990s, Philip Morris, faced with increasing pressures generated both externally, from the non-smokers' rights and public health communities, and internally, from the conflicts among its varied operating companies, seriously considered leaving the tobacco business. Discussions of this option, which occurred at the highest levels of management, focused on the changing social climate regarding tobacco and smoking that the tobacco control movement had effected. However, this option was rejected in favour of the image enhancement strategy that culminated with the recent "Altria" name change. This analysis suggests that advocacy efforts have the potential to significantly denormalise tobacco as a corporate enterprise. PMID- 12773734 TI - Development and destruction of the first state funded anti-smoking campaign in the USA. AB - BACKGROUND: Minnesota was the first state in the USA to implement a large state funded tobacco control programme (in 1985). Despite evidence of effectiveness, it was dismantled in 1993. OBJECTIVE: To describe and analyse how and why these events transpired and identify lessons for tobacco control advocates facing similar challenges in the 21st century. DESIGN: Case study based on previously secret tobacco industry documents, news reports, research reports, official documents, and interviews with health advocates and state government officials. RESULTS: Unable to defeat funding for this campaign in 1985, the tobacco industry organised groups which eliminated it later. Despite the programme's documented effectiveness, it was dismantled based on claims of fiscal crisis. These claims were not true; the real debate was what to do with the state's surplus. Health advocates failed to challenge the claim of fiscal crisis or mobilise public support for the programme. CONCLUSIONS: Simply quoting evidence that a tobacco control programme is effective does not ensure its continuing survival. Claims of fiscal crisis are an effective cover for tobacco industry efforts to dismantle successful programmes, particularly if health advocates accept these claims and fail to mobilise political pressure to defend the programme. PMID- 12773735 TI - The descriptive epidemiology of local restaurant smoking regulations in Massachusetts: an analysis of the protection of restaurant customers and workers. AB - OBJECTIVES: To describe the range of restaurant smoking regulations in the 351 cities and towns in Massachusetts, and to analyse the level of protection from secondhand smoke exposure guaranteed by these regulations. DESIGN: We obtained the local restaurant smoking regulations for each town, analysing them in terms of the protection of restaurant workers, bar workers, and adult and youth restaurant customers. MAIN OUTCOME MEASURE: The percentage of restaurant patrons and workers and bar workers who are protected from secondhand smoke exposure by the current smoking regulations in Massachusetts. RESULTS: As of June 2002, 225 towns had local restaurant smoking regulations. Of these, 69 (30.7%) do not allow smoking in restaurants, 10 (4.4%) restrict smoking to adult only restaurants, 64 (28.4%) restrict smoking to enclosed, separately ventilated areas, and 82 (36.4%) restrict smoking to areas that need not be enclosed and separately ventilated. Of the 174 towns that, at a minimum, restrict smoking to bar areas or separately ventilated areas, 35 (20.1%) allow variances. Overall, 60 towns, covering only 17.7% of the population, completely ban smoking in restaurants. As a result, 81.3% of adult restaurant customers, 81.2% of youth customers, 82.3% of restaurant workers, and 87.0% of bar workers are not guaranteed protection from secondhand smoke in restaurants. CONCLUSIONS: Despite the widespread adoption of local restaurant smoking regulations in Massachusetts, the majority of restaurant customers and workers remain unprotected from secondhand smoke exposure. In light of this, public health practitioners must stop compromising the protection of customers and workers from secondhand smoke exposure in restaurants. PMID- 12773736 TI - The impact of learning of a genetic predisposition to nicotine dependence: an analogue study. AB - OBJECTIVES: To examine the consequences of informing smokers of a genetic predisposition to nicotine dependence and of providing treatment efficacy information tailored to genetic status. DESIGN: Analogue study using four vignettes; 2 (genetic status) x 2 (whether treatment efficacy information provided) between subjects design. PARTICIPANTS: 269 British adult smokers. OUTCOME MEASURES: Preferred cessation methods and perceived control over quitting. RESULTS: Gene positive participants were significantly more likely to choose the cessation method described as effective for their genetic status, but significantly less likely to choose to use their own willpower. Providing tailored treatment information did not alter these effects. Perceived control was not significantly affected by either genetic status or information provision. CONCLUSIONS: Learning of a genetic predisposition to nicotine dependence may increase desire for effective cessation methods, but may undermine the perceived importance of willpower in stopping smoking. PMID- 12773737 TI - Smoking and gambling: a trance inducing ritual. AB - "Smoking is a powerful re-inforcement for the trance-inducing rituals associated with gambling." PMID- 12773738 TI - Three soliloquies on tobacco industry funding of university research. AB - The view on tobacco industry funding for university research, from three very differing perspectives PMID- 12773740 TI - Subsidised nicotine replacement therapy. PMID- 12773741 TI - "At Face Value": age progression software provides personalised demonstration of the effects of smoking on appearance. PMID- 12773742 TI - Smoking among workers from small companies in the Paris area 10 years after the French tobacco law. PMID- 12773744 TI - The forgotten myeloproliferative disorder: myeloid metaplasia. AB - Myelofibrosis with myeloid metaplasia is a hematologic disorder currently classified with polycythemia vera and essential thrombocythemia as a chronic myeloproliferative disease. The median age at diagnosis is 60 years, and more than 90% of patients are diagnosed after age 40 years. Clinical manifestations include massive splenomegaly, progressive anemia, profound constitutional symptoms, and extramedullary hematopoiesis. The diagnosis is confirmed by bone marrow examination after other causes of myelofibrosis are ruled out. Median survival is 5 years and causes of death include leukemic transformation. Prognosis is adversely affected by the presence of anemia (hemoglobin <10 g/dl), leukopenia or leukocytosis (white blood cells >30,000/ micro l), circulating blasts, and hypercatabolic symptoms. Conventional treatment is palliative and does not improve survival. In this regard, androgen preparations, corticosteroids, and erythropoietin are useful for the treatment of disease associated anemia. Symptomatic splenomegaly is best managed by cytoreductive therapy or surgical removal. Radiation therapy is most useful in the treatment of nonhepatosplenic extramedullary hematopoiesis. New treatment approaches include the use of thalidomide alone or in combination with prednisone and hematopoietic stem cell transplantation. PMID- 12773745 TI - Population-based pharmacoeconomic model for adopting capecitabine/docetaxel combination treatment for anthracycline-pretreated metastatic breast cancer. AB - PURPOSE: To model the cost-effectiveness of adopting capecitabine/docetaxel combination therapy in place of single-agent taxane therapy for women in the province of Ontario, Canada, receiving treatment for anthracycline-pretreated metastatic breast cancer. METHODS: Clinical effectiveness and economic data were combined in a population model, from the perspective of a universal health care system. Estimates of clinical effectiveness and medical resource utilization were derived prospectively during a phase III randomized controlled trial comparing single-agent docetaxel with capecitabine/docetaxel combination therapy. Population data were obtained from the Cancer Care Ontario Registry and provincial prescription claims data. RESULTS: During 1999-2000, 542 patients were eligible for taxane monotherapy. As capecitabine/docetaxel treatment confers a median 3-month survival benefit compared with docetaxel monotherapy, the projected survival gain in these patients was 136 life-years. The results of the cost-effectiveness analysis demonstrate that the survival benefit provided by the addition of capecitabine to single-agent docetaxel is afforded at a small incremental cost of Canadian $3,691 per life-year gained. Hospitalization costs for treatment of adverse events were less for patients receiving capecitabine/docetaxel combination therapy than for those receiving docetaxel monotherapy. The results were robust for adjustments in treatment costs and adverse effects costs. CONCLUSION: Due to its 3-month survival gain and small incremental treatment cost, capecitabine/docetaxel is judged to be a highly cost effective treatment in anthracycline-pretreated advanced breast cancer. From the perspective of the Ontario health care system, the addition of capecitabine to docetaxel in this patient population is a clinically appropriate and economically acceptable treatment strategy. PMID- 12773746 TI - A role for curative surgery in the treatment of selected patients with metastatic breast cancer. AB - Although metastatic breast cancer is widely believed to carry a grim prognosis, treatment developments over the past 25 years have greatly improved survival outcomes in these patients. In selected cases, aggressive treatment approaches may occasionally result in long-term survival of 15 years or more. This review considers the role of surgery in the treatment of single or multiple metastatic lesions restricted to one site. For each site, available literature from 1992 2002 was assessed to determine the role of surgery on survival outcomes and to determine appropriate criteria for selecting the best candidates for surgery. For lung, liver, brain, and sternum metastases, the use of surgery with or without adjuvant therapy resulted in greater median survival times and 5-year survival rates. The best candidate for surgery had no evidence of additional metastatic disease, good performance status, and a long disease-free interval after treatment of the primary tumor. Current treatment standards for breast cancer follow-up do not include imaging studies other than mammography. The addition of chest x-rays as part of routine follow-up should be considered as a cost effective approach for early assessment of metastases to the lung or sternum that may be appropriate for surgical excision. PMID- 12773747 TI - Clinical application of cDNA microarrays in oncology. AB - DNA microarrays represent an important new tool to analyze human tissues. The technology enables investigators to measure the expression of several thousand mRNA species simultaneously in a biological specimen. This process, called transcriptional profiling, represents a technological breakthrough in the analysis of biological specimens. It may be used to screen for individual genes that are differentially expressed between normal and diseased tissues in the hope of finding novel targets for drug development or finding new single-gene markers of clinical outcome. Microarrays are also applied to learn about the complex biology of cancer by simultaneously monitoring interactions between hundreds of genes during experimental conditions in vitro or during therapy in vivo. Analysis of gene expression patterns may also be used as a classification tool to sort cancer into various clinically relevant subgroups that is not currently possible with other methods. The first clinically important applications of this technology will likely be its use as a tool to refine diagnosis and improve the accuracy of predictions of prognosis and response to therapy. DNA microarrays in several "proof-of-principle" experiments have demonstrated that they can predict important clinical outcomes, including outcomes that cannot currently be predicted with other methods, but the true clinical utility and the limits of this exciting new technology are yet to be established. This paper reviews the current methodology and applications of this technique as they relate to clinical oncology. PMID- 12773748 TI - Management of high-risk populations with locally advanced prostate cancer. AB - Prostate cancer that extends beyond the confines of the prostatic gland on clinical and/or radiographic assessment, without evidence of lymph node or distant metastases, is regarded as locally advanced. The locally advanced prostate cancer patient population consists of a heterogeneous group of men, some of whom have tumors that may be amenable to primary curative intent with local definitive therapy associated with acceptable long-term cancer control rates. In order to optimally manage this group of patients, it is important to be able to recognize who is at a high risk of tumor recurrence after primary local therapy. In this brief review, we discuss the factors that contribute to the prediction of high risk in populations with locally advanced disease and the treatment options available. PMID- 12773749 TI - Chemotherapy options for the elderly patient with advanced non-small cell lung cancer. AB - Combination chemotherapy has been shown to improve overall survival compared with best supportive care in patients with advanced non-small cell lung cancer (NSCLC). The survival advantage is modest and was initially demonstrated with cisplatin-containing regimens in a large meta-analysis of randomized trials reported in 1995. Newer chemotherapy combinations have been shown to be better tolerated than older cisplatin-based combinations, and some trials have also shown greater efficacy and survival benefits with these newer combinations. Combination chemotherapy is, therefore, the currently accepted standard of care for patients with good performance statuses aged less than 70 years with advanced NSCLC. However, there are limited data from clinical trials to support the use of combination chemotherapy in elderly patients over 70 years of age with advanced NSCLC. Subgroup analyses of large randomized phase III trials suggest that elderly patients with good performance statuses do as well as younger patients treated with combination chemotherapy. There are few randomized trials reported that evaluate chemotherapy in patients aged greater than 70 years only. Based on data from trials performed by an Italian group, single-agent vinorelbine has been shown to have significant activity in elderly patients with advanced NSCLC and to be well tolerated by those patients with Eastern Cooperative Oncology Group performance statuses of two or less, with associated improvements in measures of global health. PMID- 12773750 TI - Advances in the diagnosis and treatment of neuroblastoma. AB - Neuroblastoma, a childhood neoplasm arising from neural crest cells, is characterized by a diversity of clinical behavior ranging from spontaneous remission to rapid tumor progression and death. To a large extent, outcome can be predicted by the stage of disease and the age at diagnosis. However, the molecular events responsible for the variability in response to treatment and the rate of tumor growth remain largely unknown. Over the past decade, transformation linked genetic changes have been identified in neuroblastoma tumors that have contributed to the understanding of tumor predisposition, metastasis, treatment responsiveness, and prognosis. The Children's Oncology Group recently developed a Neuroblastoma Risk Stratification System that is currently in use for treatment stratification purposes, based on clinical and biologic factors that are strongly predictive of outcome. This review discusses the current risk-based treatment approaches for children with neuroblastoma and recent advances in biologic therapy. PMID- 12773751 TI - The molecular perspective: ubiquitin and the proteosome. PMID- 12773753 TI - Role of double-stranded RNA-activated protein kinase R (PKR) in deoxynivalenol induced ribotoxic stress response. AB - Trichothecene mycotoxins and other protein synthesis inhibitors activate mitogen activated protein kinase (MAPKs) via a mechanism that has been termed the "ribotoxic stress response." MAPKs are believed to mediate the leukocyte apoptosis that is observed following experimental exposure to these chemical agents in vitro and in vivo. The purpose of this research was to test the hypothesis that double-stranded, RNA-activated protein kinase R (PKR) is a critical upstream mediator of the ribotoxic stress response induced by the trichothecene deoxynivalenol (DON) and other translational inhibitors. DON was found to readily induce phosphorylation of JNK 1/2, ERK 1/2, and p38 in the murine macrophage RAW 264.7 cell line, within 5 min of culture addition, in a concentration-dependent fashion. Effects were maximal from 15 to 30 min and lasted up to 6 h. The translational inhibitors anisomycin and emetine also had similar effects when added to cultures at equipotent concentrations to DON. DON rapidly activated PKR within 1 to 5 min, as evidenced by autophosphorylation and by phosphorylation of eukaryotic initiation factor 2alpha (eIF2alpha). Interestingly, the latter effect was associated with rapid degradation of eIF2alpha. Pretreatment of RAW 264.7 cells with two inhibitors of PKR, 2 aminopurine (2-AP) or adenine (Ad), markedly impaired MAPK phosphorylation in RAW 264.7 cells according to the following rank order JNK>p38>ERK. The capacity of DON to induce MAPK phosphorylation was also markedly suppressed in a stable transformant of the human promonocytic U-937 cell line containing an antisense PKR expression vector. This suppression followed a rank order of JNK>p38>ERK in this PKR-deficient cell line when compared to control cells transfected with vector only. Apoptosis induction by DON and two other translational inhibitors, anisomycin and emetine, was almost completely abrogated in PKR-deficient cells. Together, the results indicate that PKR plays a critical upstream role in the ribotoxic stress response inducible by translational inhibitors. PMID- 12773754 TI - Immune changes during acute cold/restraint stress-induced inhibition of host resistance to Listeria. AB - Experiments were conducted to delineate the cellular changes modulated by acute cold/restraint stress (ACRS), a physical and psychological stressor, in response to a Listeria monocytogenes(LM) infection. In addition to wild type (WT) BALB/c mice, CD4-deficient (CD4-/-) BALB/c mice, which have no effective adaptive immunity, were used to determine the involvement of adaptive versus innate immunity. ACRS-induced suppression of host resistance to LM was not observed in CD4-/- mice, suggesting the involvement of CD4+T cells in the acute cold/restraint stress (ACRS)-induced inhibition. The in vivo splenic leukocyte phenotypes and activities of WT BALB/c mice after infection and in vitro lymphocyte responses to heat-killed LM (HKLM) also were examined. There were no significant differences in the numbers of splenic T and B lymphocytes, natural killer cells, macrophages, or neutrophils between nonstressed and ACRS-treated WT mice. However, higher levels of activated T cells and non-T lymphocytes were observed in the ACRS-treated mice; beta-adrenergic receptor (beta-ADR) antagonists (propranolol and atenolol) eliminated these elevated levels of activation, as well as the ACRS-induced suppression of host resistance. ACRS and control mice also had equivalent activation of macrophages. With in vitro HKLM stimulation, splenocytes from ACRS-treated mice produced significantly higher levels of IFNgamma and slightly higher levels of IL-6 in comparison with the nonstressed mice, although equivalent levels of lymphocyte proliferation were obtained. Additionally, ACRS-treated mice showed comparable elevation of serum nitric oxide after infection, indicating macrophage bactericidal activity similar to nonstressed mice. Thus, it appears that ACRS inhibits host resistance through regulatory CD4+ T cells and/or effector cell functions downstream of CD4+ T cell activation, as well as through beta-ADR signaling, in that blockage of these receptors appears to aid host defenses by means other than elevation of helper T cell activity. Because CD4 T cell deficiency and beta-ADR blockage produced equivalent effects, beta-ADR+ CD4+ T cells may have a negative role on host defenses after ACRS. PMID- 12773755 TI - Species-dependent variations in the in vitro myotoxicity of death adder (Acanthophis) venoms. AB - Based on early studies on Acanthophis antarcticus (common death adder) venom, it has long been thought that death adder snake venoms are devoid of myotoxicity. However, a recent clinical study reported rhabdomyolysis in patients following death adder envenomations, in Papua New Guinea, by a species thought to be different to A. antarcticus. Subsequently, a myotoxic phospholipase A2 component was isolated from A. rugosus (Irian Jayan death adder) venom. The present study examined the venoms of A. praelongus (northern), A. pyrrhus (desert), A. hawkei (Barkly Tableland), A. wellsi (black head), A. rugosus, A. sp. Seram and the regional variants of A. antarcticus for in vitro myotoxicity. Venoms (10-50 microg/ml) were examined for myotoxicity using the chick directly (0.1 Hz, 2 ms, supramaximal V) stimulated biventer cervicis nerve-muscle preparation. A significant contracture of skeletal muscle and/or inhibition of direct twitches were considered signs of myotoxicity. This was confirmed by histological examination. All venoms displayed high phospholipase A2 activity. The venoms (10 50 microg/ml) of A. sp. Seram, A. praelongus, A. rugosus,and A. wellsi caused a significant inhibition of direct twitches and an increase in baseline tension compared to the vehicle (n=4-6; two-way ANOVA, p<0.05). Furthermore, these venoms caused dose-dependent morphological changes in skeletal muscle. In contrast, the venoms (10-50 microg/ml; n=3-6) of A. hawkei, A. pyrrhus, and regional variants of A. antarcticus were devoid of myotoxicity. Prior incubation (10 min) of CSL death adder antivenom (5 U/ml) prevented the myotoxicity caused by A. sp. Seram, A. praelongus, A. rugosus, and A. wellsi venoms (50 microg/ml; n=4-7). In conclusion, clinicians may need to be mindful of possible myotoxicity following envenomations by A. praelongus, A. rugosus, A. sp. Seram, and A. wellsi species. PMID- 12773756 TI - Comparative scoring of micronucleated reticulocytes in rat peripheral blood by flow cytometry and microscopy. AB - A flow cytometric technique for scoring the incidence of micronucleated reticulocytes in rat peripheral blood was compared to a standard microscopy-based procedure. For these studies, groups of five male Sprague-Dawley rats were treated with vehicle or a broad range of chemical genotoxicants: 6-thioguanine, N methyl-N'-nitro-N-nitrosoguanidine, vincristine, methylaziridine, acetaldehyde, methyl methanesulfonate, benzene, monocrotaline, and azathioprine. Animals were treated once a day for up to 2 days, and peripheral blood was collected between 24 and 48 h after the final administration. These samples were processed for flow cytometric scoring and microscopy-based analysis using supravital acridine orange staining, and the percentage of reticulocytes and micronucleated reticulocytes was determined for each sample. The resulting data demonstrate good agreement between these scoring methodologies, although careful execution of the flow cytometric method was found to enhance the micronucleus assay by reducing both scoring time and scoring error. These data add further support to the premise that the peripheral blood compartment of rats can be used effectively to detect genotoxicant-induced micronuclei. PMID- 12773757 TI - Gene expression analysis of the acute phase response using a canine microarray. AB - The safety of pharmaceuticals is typically assessed in the dog and rat prior to investigation in humans. As a result, a greater understanding of adverse effects in these preclinical testing species would improve safety assessment. Despite this need, there is a lack of tools to examine mechanisms and identify biomarkers in the dog. To address this issue, we developed an Affymetrix-based oligonucleotide microarray capable of monitoring the expression of thousands of canine genes in parallel. The custom canine array contains 22,774 probe sets, consisting of 13,729 canine and 9045 human-derived probe sets. To improve cross species hybridization with human-derived probes, the detection region was moved from the variable 3' UTR to the more homologous coding region. Testing of this strategy was accomplished by comparing hybridization of naive dog liver RNA to the canine array (coding region design) and human U133A array (standard 3' design). Although raw signal intensity was greater with canine-specific probe sets, human-derived probes detected the expression of additional liver transcripts. To assess the ability of this tool to detect differential gene expression, the acute phase response was examined in beagle dogs given lipopolysaccharide (LPS). Hepatic gene expression 4 and 24 h post-LPS administration was compared to gene expression profiles of vehicle-treated dogs (n=3/group). Array data was consistent with an acute inflammatory response, with transcripts for multiple cytokines and acute phase proteins markedly induced 4 h after LPS challenge. Robust changes in the expression of transcripts involved with glucose homeostasis, biotransformation, and extracellular matrix remodeling were observed 24 h post-dose. In addition, the canine array identified several potential biomarkers of hepatic inflammation. Strong correlations were found between gene expression data and alterations in clinical chemistry parameters such as serum amyloid A (SAA), albumin, and alkaline phosphatase (ALP). In summary, this new genomic tool successfully detected basal canine gene expression and identified novel aspects of the acute phase response in dog that shed new light on mechanisms underlying inflammatory processes. PMID- 12773758 TI - Evaluation of the Tg.AC assay: specificity testing with three noncarcinogenic pharmaceuticals that induce selected stress gene promoters in vitro and the inhibitory effects of solvent components. AB - Understanding the strengths and limitations of alternative models, such as the Tg.AC assay, for evaluation of the potential carcinogenicity of pharmaceuticals requires assessment of assay specificity through studies that specifically target biologically active compounds that are known to not be carcinogens in rodents. To identify drugs that might provoke a false positive response in the Tg.AC assay, we screened pharmaceuticals for in vitro induction of the gadd153 promoter and the zeta-globin promoter. We have previously found a high correlation between induction of the gadd153 promoter in HepG2 cells and activity in the Tg.AC assay. The three drugs selected through screening 99 noncarcinogenic pharmaceuticals were amiloride, dipyridamole, and pyrimethamine. A 26-week skin paint study was conducted in hemizygous Tg.AC mice with the three drugs at two doses selected by a 4-week dose range finding study. Evidence of systemic toxicity was observed in animals dosed chronically with pyrimethamine or amiloride, but no skin papillomas were observed in mice treated with amiloride, dipyridamole, or pyrimethamine for 26 weeks. All male mice and 80% of female mice treated with 12-O tetradecanoylphorbol-13-acetate (TPA) in acetone developed a maximal tumor burden. However, mice treated with TPA in a vehicle containing 2.4% DMSO had greatly reduced incidences of papillomas. In summary, the correct negative response was shown in the Tg.AC assay for three noncarcinogenic pharmaceuticals, which adds further favorable evidence of appropriate specificity of this model system. However, vehicle composition must be carefully selected because the outcome of this assay can be confounded by certain commonly used solvents. PMID- 12773759 TI - The long (LINEs) and the short (SINEs) of it: altered methylation as a precursor to toxicity. AB - Although once thought of as "junk" DNA, the importance of interspersed elements in the genome has become increasingly appreciated in recent years. In a broad sense these are collectively referred to as transposable elements, which encompass both transposons and retrotransposons. The latter include long interspersed nuclear elements (LINEs) and short interspersed nuclear elements (SINEs). Expression of these elements leads to genetic instability. Therefore, it is important that they remain transcriptionally silenced, and DNA methylation plays a key role in this regard. A framework for understanding the possible interplay between altered DNA methylation, an epigenetic change, and mutational events is presented. A case is made as to how retrotransposable elements, specifically LINEs and SINEs, are likely to emerge as key players in furthering our understanding of mechanisms underlying a variety of toxicities, including carcinogenesis but not limited to this endpoint. PMID- 12773760 TI - Nickel-induced histone hypoacetylation: the role of reactive oxygen species. AB - The carcinogenicity of specific insoluble nickel compounds is mainly due to their intracellular generation of Ni2+ ion and its suppression on gene transcription, while the inhibition of Ni2+ on histone acetylation plays an important role in the suppression or silencing of genes. Recent studies on Ni2+ and histone H4 acetylation suggest that Ni2+ inhibits the acetylation of histone H4 through binding with its N-terminal histidine-18. It is well known that bound Ni2+ readily produces reactive oxygen species (ROS) in vivo, a critical factor inversely related with the occurrence of resistance of mammalian cells to Ni2+. Thus, we tried to find the possible role of ROS in the induction of Ni2+ on histone acetylation in the present study. We found that a high concentration of Ni2+ (no less than 600 microM) caused a significant decrease of histone acetylation in human hepatoma cells. This inhibition was shown to result mainly from the influence of Ni2+ on the overall histone acetyltransferase (HAT) activity indicated by the histone acetylation assay with the presence of a specific histone deacetylase (HDAC) inhibitor, trichostatin A (TSA). The in vitro HAT and HDAC assays further confirmed this result. At the same time, we found that the exposure of hepatoma cells to Ni2+ generated ROS. Coadministration of hydrogen peroxide with Ni2+ generated more ROS and more histone acetylation inhibition. Addition of the antioxidants 2-mercaptoethanol (2-ME) at 2 mM or N acetyl-cysteine (NAC) at 1 mM, with Ni2+ together, completely suppressed ROS generation and significantly diminished the induced histone hypoacetylation. The data presented here prove that the ROS generation plays a role in the inhibition of histone acetylation, and, hence, the gene suppression and carcinogenesis caused by Ni2+ exposure, providing a new door for us to continuously understand the mechanism of ROS in the carcinogenicity of Ni2+ and the resistance of mammalian cells to Ni2+. PMID- 12773761 TI - Carcinogenicity of inhaled vanadium pentoxide in F344/N rats and B6C3F1 mice. AB - Vanadium pentoxide (V2O5) is a slightly soluble compound found in airborne particle emissions from metallurgical works and oil and coal burning. Because the carcinogenic potential of V2O5 was not known, F344/N rats and B6C3F1 mice (N=50/sex/species) were exposed to V2O5 at concentrations of 0, 0.5 (rats only), 1, 2, or 4 (mice only) mg/m3, by whole-body inhalation for 2 years. The survival and body weights of rats were minimally affected by exposure to V2O5. The survival and body weights of male mice exposed to 4 mg/m3 and body weights of all exposed groups of female mice were lower than the controls. Alveolar/bronchiolar (A/B) neoplasms occurred in male rats exposed to 0.5 and 2 mg/m3 at incidences exceeding the National Toxicology Program (NTP) historical control ranges. A marginal increase in A/B neoplasms was also observed in female rats exposed to 0.5 mg/m3. Increases in chronic inflammation, interstitial fibrosis, and alveolar and bronchiolar hyperplasia/metaplasia and squamous metaplasia were observed in exposed male and female rats. A/B neoplasms were significantly increased in all groups of exposed mice. As with rats, increases in chronic inflammation, interstitial fibrosis, and alveolar and bronchiolar epithelial hyperplasia were observed in mice exposed to V2O5. Thus, V2O5 exposure was a pulmonary carcinogen in male rats and male and female mice. The marginal tumor response in the lungs of female rats could not be attributed conclusively to exposure to V2O5. These responses were noted at and slightly above the OSHA permissible occupational exposure limit of 0.5 mg/m3 (dust) (National Institute for Occupational Safety and Health, NIOSH Pocket Guide to Chemical Hazards, U.S. Department of Health and Human Services, Washington, DC, 1997, p. 328). PMID- 12773762 TI - Pyruvate attenuates myoglobin in vitro toxicity. AB - Myoglobinuria is a complication of crush injury as well as substance abuse. This study examined whether pyruvate modified myoglobin in vitro renal toxicity. Renal slices from Fischer-344 rats were incubated for 120 min with 0-12 mg/ml myoglobin. In an initial study, gluconeogenesis was stimulated by the addition of 10 mM pyruvate during the final 30 min. In all other studies, renal slices were incubated with myoglobin in the presence of 0 or 10 mM pyruvate for 120 min. Myoglobin increased lactate dehydrogenase (LDH) release and this was not modified by the presence of pyruvate for the last 30 min of the incubation. Myoglobin toxicity was reduced by coincubation of myoglobin with pyruvate for 120 min. LDH leakage was increased 1.2-, 1.7-, and 1.8-fold above control by 4, 10, and 12 mg/ml myoglobin, compared to 1.2, 1.3, and 1.3 fold in slices coincubated with 10 mM pyruvate, respectively. Myoglobin diminished adenosine triphosphate (ATP) levels but pyruvate maintained a 5x higher level of ATP within the slices. Glucose (10 mM) provided protection only for the low concentration (4 mg/ml) of myoglobin. Myoglobin induced oxidative stress while pyruvate prevented the rise in lipid peroxidation and glutathione disulfides by myoglobin. Myoglobin diminished total glutathione levels in pyruvate-treated tissue, but glutathione levels remained higher than tissues incubated in the absence of pyruvate. These results indicate that pyruvate reduced toxicity by preventing oxidative stress and via a supply of an energy substrate. PMID- 12773763 TI - Metabolic activation of 2-hydroxyamino-1-methyl-6-phenylimidazo[4,5-b]pyridine in Syrian hamsters congenic at the N-acetyltransferase 2 (NAT2) locus. AB - 2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) is a heterocyclic amine carcinogen prevalent in the human diet. To exert its mutagenic and carcinogenic effects, PhIP undergoes bioactivation to N-hydroxy-PhIP followed by O esterification via cytosolic acetyltransferases or sulfotransferases to form DNA adducts. We investigated the role of cytosolic acetyltransferases and sulfotransferases and the role of the N-acetyltransferase 2 genetic polymorphism on PhIP DNA-adduct levels in a congenic Syrian hamster model. DNA adduct levels were detected in all hepatic and extrahepatic tissues tested following administration of PhIP (4x100 mg/kg) or N-hydroxy-PhIP (1x50 mg/kg), with the highest levels in pancreas. DNA-adduct levels were higher in the gastrointestinal tract of rapid and slow acetylator hamsters administered N-hydroxy-PhIP. N hydroxy-PhIP O-acetyltransferase and O-sulfotransferase activities were detected in most hepatic and extrahepatic cytosols derived from rapid and slow acetylator congenic hamsters. N-hydroxy-PhIP O-acetyltransferase activity was significantly higher (p<0.05) in liver, small intestine, and esophagus in rapid than in slow acetylator congenic hamsters. N-hydroxy-PhIP O-acetyltransferase activities correlated significantly with N-acetyltransferase 2 activities across tissues in rapid (r=0.83; p=0.0004) but not in slow (r=0.46; p=0.1142) acetylator congenic hamsters, suggesting catalysis primarily by NAT2 in rapid acetylators but NAT1 in slow acetylators. N-hydroxy-PhIP O-sulfotransferase activities did not vary with acetylator genotype. DNA-adduct levels following administration of PhIP or N hydroxy-PhIP did not correlate with either N-hydroxy-PhIP O-acetyltransferase or O-sulfotransferase catalytic activities. PMID- 12773764 TI - Suppressive effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on the high affinity antibody response in C57BL/6 mice. AB - In the humoral immune response to an invasion of foreign antigens, B cells differentiate into low-affinity antibody-forming cells (AFCs) that mainly secrete IgM or, through germinal center (GC) formation, into high-affinity AFCs that secrete IgG-class antibodies with a higher affinity for the antigen. Previous studies have established the suppressive effects of 2,3,7,8-tetrachlorodibenzo-p dioxin (TCDD) on low-affinity antibody responses to antigens. However, whether and how TCDD affects the high-affinity antibody response to antigens has not yet been clarified. In this paper we investigate the effects of TCDD on GC formation, high-affinity AFC generation, and high-affinity antibody production in the primary humoral immune response. C57BL/6 mice were orally administered 0 or 20 microg/kg of TCDD and subsequently immunized with alum-precipitated ovalbumin (OVA) on day 0. Then the GC formation in the spleen and OVA-specific antibodies in the plasma, was evaluated until day 14 postimmunization. TCDD exposure reduced the production of OVA-specific IgG1 on days 10 and 14. GC formation in the spleen was also suppressed by TCDD exposure, and the suppression persisted from day 7 until day 14. In TCDD-administered mice, on day 7, cellular proliferation in the GCs was significantly suppressed, although apoptosis was not markedly affected. In order to measure high-affinity antibody and high-affinity AFCs, the mice were administered TCDD followed by immunization with alum-precipitated (4-hydroxy-3 nitrophenyl) acetyl linked to chicken gamma-globulin (NP-CG). The frequency of high-affinity NP-specific AFCs that bind to low-haptenated antigen was clearly shown to be reduced in the spleen on days 10 and 14. Furthermore, the high affinity anti-NP IgG1 levels on days 10 and 14 postimmunization were significantly reduced by TCDD exposure. Taken together, the results of this paper demonstrate that TCDD exposure inhibits the generation of high-affinity AFCs and high-affinity antibody production during the primary humoral immune response and suggest that these alterations were caused by the suppression of antigen responding B-cell proliferation induced by TCDD during GC formation. PMID- 12773765 TI - Effects of in utero tributyltin chloride exposure in the rat on pregnancy outcome. AB - Tributyltin, an organotin, is ubiquitous in the environment. The consumption of contaminated marine species leads to human dietary exposure to this compound. Tributyltin is an endocrine disruptor in many wildlife species and inhibits aromatase in mammalian placental and granulosa-like tumor cell lines. We investigated the effects of tributyltin chloride exposure on pregnancy outcome in the Sprague-Dawley rat. Timed pregnant rats were gavaged either with vehicle (olive oil) or tributyltin chloride (0.25, 2.5, 10, or 20 mg/kg) from days 0-19 or 8-19 of gestation. On gestational day 20, dams were sacrificed, and pregnancy outcome was determined. Tributyltin and its metabolites (dibutyltin, monobutyltin) were measured in maternal blood by gas chromatography. Both tributyltin and dibutyltin were present in maternal blood at approximately equal concentrations, whereas monobutyltin contributed minimally to total organotins. Organotin concentrations increased in a dose-dependent pattern in dams, independent of the window of exposure. Tributyltin chloride administration significantly reduced maternal weight gain only at the highest dose (20 mg/kg); a significant increase in post-implantation loss and decreased litter sizes, in addition to decreased fetal weights, was observed in this group. Tributyltin chloride exposure did not result in external malformations, nor was there a change in sex ratios. However, exposure to 0.25, 2.5, or 10 mg/kg tributyltin chloride from gestation days (GD) 0-19 resulted in a significant increase in normalized anogenital distances in male fetuses; exposure from days 8-19 had no effect. There was a dramatic increase in the incidence of low weight (< or =0.75 of the mean) fetuses after exposure to 20 mg/kg tributyltin chloride. Delayed ossification of the fetal skeleton was observed after in utero exposure to either 10 mg/kg or 20 mg/kg tributyltin chloride. Serum thyroxine and triiodothyronine levels were reduced significantly in dams exposed to 10 and 20 mg/kg tributyltin chloride throughout gestation; in dams treated with tributyltin from GD 8-19, serum thyroxine concentrations, but not triiodothyronine, were significantly decreased at both the 2.5 and 10 mg/kg exposures. Thus, maternal thyroid hormone homeostasis may be important in mediating the developmental toxicity of organotins. PMID- 12773766 TI - The coagulation system contributes to synergistic liver injury from exposure to monocrotaline and bacterial lipopolysaccharide. AB - Coexposure to a noninjurious dose of bacterial lipopolysaccharide (LPS; 7.4 x 106 EU/kg) and a nontoxic dose of the food-borne toxin monocrotaline (MCT; 100 mg/kg) leads to synergistic hepatotoxicity in Sprague-Dawley rats. Inflammatory factors, such as Kupffer cells (KCs), tumor necrosis factor-alpha (TNF)-alpha, and neutrophils (polymorphonuclear leukocytes; PMNs), are critical to the pathogenesis. Inasmuch as activation of the coagulation system and sinusoidal endothelial cell (SEC) injury precede hepatic parenchymal cell (HPC) injury, and since fibrin deposition occurs within liver lesions, the coagulation system might be a critical component of injury. In this study, this hypothesis is tested, and the interdependence of the coagulation system and inflammatory factors is explored. Administration of the anticoagulants heparin or warfarin to MCT/LPS cotreated animals attenuated HPC and SEC injury. Morphometric analysis revealed that anticoagulant treatment significantly reduced the area of centrilobular and midzonal lesions. Heparin treatment also reduced fibrin deposition in these regions. Furthermore, anticoagulant treatment decreased hepatic PMN accumulation but did not affect plasma TNF-alpha concentration. Neither KC inactivation nor TNF-alpha depletion prevented activation of the coagulation system. PMN depletion, however, prevented coagulation system activation, suggesting that PMNs are needed for this response. These results provide evidence that the coagulation system and its interplay with PMNs are important in the pathogenesis of MCT/LPS induced liver injury. PMID- 12773767 TI - Effects of in utero exposure to finasteride on androgen-dependent reproductive development in the male rat. AB - Finasteride is a specific inhibitor of type II 5alpha-reductase, the enzyme that converts testosterone (T) to the more potent androgen receptor agonist dihydrotestosterone (DHT). In utero exposure to androgen receptor antagonists and T biosynthesis inhibitors have induced permanent effects on androgen-sensitive end points such as anogenital distance (AGD), nipple retention, and malformations of the male rat reproductive tract. The objectives of this study were to (1) characterize the dose response of finasteride-mediated alterations in androgen dependent developmental end points, (2) determine whether prenatal exposure to finasteride permanently decreases AGD or results in nipple retention, and (3) evaluate whether AGD or nipple retention is predictive of adverse alterations in the male reproductive tract. Pregnant Crl:CD(SD)BR rats (n=5-6/group) were gavaged with either vehicle or finasteride at 0.01, 0.1, 1.0, 10, or 100 mg/kg/day on gestation days 12 to 21. All male offspring were monitored individually until necropsy on postnatal day (PND) 90. The present study design has been used previously for other antiandrogens and is sensitive to perturbations of the male rat reproductive tract. Decreases in AGD on PND 1 and increases in areolae-nipple retention on PND 13 were significantly different from controls in all finasteride-exposed male rats. Finasteride-induced changes in AGD and nipple retention were permanent in male rats exposed to finasteride at and above 0.1 mg/kg/day. On PND 90, dorsolateral and ventral prostate lobes were absent in 21 to 24% of rats exposed to 100 mg/kg/day finasteride and weighed significantly less at and above 10 mg/kg/day. In the highest dose group, 73% of animals had ectopic testes, much higher than previously reported. The most sensitive malformation other than decreased AGD and nipple retention was the dose dependent increase in hypospadias. The lowest observed adverse effect level (LOAEL) for finasteride-induced permanent effects in this study was 0.1 mg/kg/day based on permanent changes in AGD and nipple retention. Finasteride-induced changes in AGD and retention of nipples were highly predictive of hypospadias, ectopic testes, and prostate malformations even though some animals with retained nipples or decreased AGD may not have had other reproductive tract malformations. In summary, prenatal exposure to finasteride specifically inhibited DHT-mediated development with little to no change in T-mediated development. PMID- 12773768 TI - Thimerosal induces DNA breaks, caspase-3 activation, membrane damage, and cell death in cultured human neurons and fibroblasts. AB - Thimerosal is an organic mercurial compound used as a preservative in biomedical preparations. Little is known about the reactions of human neuronal and skin cells to its micro- and nanomolar concentrations, which can occur after using thimerosal-containing products. A useful combination of fluorescent techniques for the assessment of thimerosal toxicity is introduced. Short-term thimerosal toxicity was investigated in cultured human cerebral cortical neurons and in normal human fibroblasts. Cells were incubated with 125-nM to 250-microM concentrations of thimerosal for 45 min to 24 h. A 4', 6-diamidino-2-phenylindole dihydrochloride (DAPI) dye exclusion test was used to identify nonviable cells and terminal transferase-based nick-end labeling (TUNEL) to label DNA damage. Detection of active caspase-3 was performed in live cell cultures using a cell permeable fluorescent caspase inhibitor. The morphology of fluorescently labeled nuclei was analyzed. After 6 h of incubation, the thimerosal toxicity was observed at 2 microM based on the manual detection of the fluorescent attached cells and at a 1-microM level with the more sensitive GENios Plus Multi-Detection Microplate Reader with Enhanced Fluorescence. The lower limit did not change after 24 h of incubation. Cortical neurons demonstrated higher sensitivity to thimerosal compared to fibroblasts. The first sign of toxicity was an increase in membrane permeability to DAPI after 2 h of incubation with 250 microM thimerosal. A 6-h incubation resulted in failure to exclude DAPI, generation of DNA breaks, caspase-3 activation, and development of morphological signs of apoptosis. We demonstrate that thimerosal in micromolar concentrations rapidly induce membrane and DNA damage and initiate caspase-3-dependent apoptosis in human neurons and fibroblasts. We conclude that a proposed combination of fluorescent techniques can be useful in analyzing the toxicity of thimerosal. PMID- 12773769 TI - Glutathione depletion exacerbates methylenedianiline toxicity to biliary epithelial cells and hepatocytes in rats. AB - Methylenedianiline (DAPM) initially injures epithelial cells of major bile ducts, which is followed by cholestasis, cholangitis, and hepatocellular damage. This pattern of biliary injury resembles that produced by alpha-naphthylisothiocyanate (ANIT), a classic bile duct toxicant. Our goal was to determine whether prior depletion of hepatic total glutathione (GSx), a condition reported to protect against biliary tract injury by ANIT, would also protect against DAPM-induced bile duct injury. A new protocol for extensive, sustained depletion of GSx was established. We found that administration of 1-bromoheptane followed 1 h later by buthionine sulfoximine resulted in an approximately 96% depletion of hepatic GSx that persisted through 6 h without biochemical or morphological signs of hepatic injury. Treatment of rats with a minimally hepatotoxic dose of DAPM (without GSx depletion) produced at 6 h injury similar to previous studies: moderate oncosis of biliary epithelial cells (BEC), mild edema of portal triads, and increases in glutathione S-transferase (GST) activities without alterations in hepatic GSx/glutathione disulfide (GSSG), coenzyme A (CoASH)/coenzyme A-glutathione disulfide (CoASSG), or thiobarbituric acid-reactive substances (TBARS). In contrast, DAPM treatment of GSx-depleted rats produced severe oncosis of BEC, marked inflammatory and edematous alterations to portal tracts, and oncosis/apoptosis in scattered hepatocytes. The observed acceleration and enhancement of DAPM-induced liver injury by GSx depletion was associated with a concurrent sevenfold increase in hepatic CoASSG and a fourfold decrease in the ratio of CoASH to CoASSG, compounds presumably localized to mitochondria and a purported index of mitochondrial thiol/disulfide status. These results indicate that: (1) GSx depletion exacerbates BEC and hepatocellular injury induced by DAPM, and (2) the mechanism by which DAPM causes liver injury is likely different from that of the classic bile duct toxicant, ANIT. PMID- 12773770 TI - Gene expression patterns as potential molecular biomarkers for malignant transformation in human keratinocytes treated with MNNG, arsenic, or a metal mixture. AB - In previous studies, treatment with 1-methyl-3-nitro-1-nitrosoguanidine (MNNG) enhanced malignant transformation of immortal human epidermal (RHEK-1) keratinocytes. In contrast, arsenic (As) alone or in a mixture of As, cadmium (Cd), chromium (Cr), and lead (Pb) inhibited this process. Microarray analysis showed unique gene expression patterns in RHEK-1 exposed to MNNG, As, or the metal mixture. From this analysis, we have selected 16 genes potentially involved in the enhancement or inhibition of transformation. These 16 genes, nine (IFN inducible protein 9-27, MAA A32, CCLB protein, integrin beta4, XRCC1, K8, K18, MT3, MAPKK6) of which were altered in a chemical-specific manner and seven (MIC1, bikunin, MTS1, BMP4, RAD23A, DOC2, vimentin) of which were commonly affected by the MNNG and As or mixture treatments, were examined for expression in detail by real-time RT-PCR. Qualitatively, both microarray and real-time RT-PCR analyses gave comparable results for 15 of 16 genes, i.e., genes were consistently induced or suppressed under the different treatment regimens when measured by either technique. Of the seven genes altered in their expression by multiple chemical treatments, five showed patterns consistent with a role in the transformation process, i.e., they were oppositely regulated in MNNG-transformed RHEK-1 cells (designated as OM3) as compared to the nonmalignant As- and mixture-exposed cells. Through time-course studies, we also identified markers whose expression correlates with acquisition of transformation-associated characteristics in OM3. Identification of a battery of genes altered during progressive transformation of RHEK-1 should aid in developing a mechanistic understanding of this process, as well as strengthening the utility of these genes as biomarkers. PMID- 12773771 TI - Growth and angiogenesis are inhibited in vivo in developing tissues by pyrazine and its derivatives. AB - Sidestream cigarette smoke solution was previously screened to identify the groups of chemicals in smoke that inhibit growth and angiogenesis in the chick chorioallantoic membrane (CAM). Pyrazine and several pyrazine derivatives were identified as a major chemical group in this screen. In the current study, purified pyrazine and six pyrazine derivatives identified in the screen were tested in dose response experiments to measure their effects on CAM growth, embryo growth, and angiogenesis. Chemicals or control medium were placed on CAMs in ovo on day 5 of development, and results were evaluated on day 6. Of the chemicals tested, pyrazine was the most potent and inhibited both CAM and embryo growth at picomolar doses. 2-Ethylpyrazine and 2,3-dimethylpyrazine were inhibitory at nanomolar doses. Inhibition of growth by pyrazine was correlated with inhibition of DNA synthesis. The pattern of blood vessel development in CAMs was disturbed by micromolar doses of pyrazine and 2,3-dimethylpyrazine. Migration of mesodermal blood vessels to the ectoderm of CAMs and their subsequent differentiation into the capillary plexus was impaired by nanomolar doses of pyrazine. In summary, these data show that pyrazine and some of its derivatives inhibit growth and certain processes important in angiogenesis at very low doses. Since pyrazine and some of its derivatives are considered safe food additives, further toxicological testing of pyrazine, in particular on developing tissues, should be done to fully evaluate its safety as a consumer product additive. PMID- 12773772 TI - Exposure to perfluorooctane sulfonate during pregnancy in rat and mouse. II: postnatal evaluation. AB - The postnatal effects of in utero exposure to perfluorooctane sulfonate (PFOS, C8F17SO3-) were evaluated in the rat and mouse. Pregnant Sprague-Dawley rats were given 1, 2, 3, 5, or 10 mg/kg PFOS daily by gavage from gestation day (GD) 2 to GD 21; pregnant CD-1 mice were treated with 1, 5, 10, 15, and 20 mg/kg PFOS from GD 1 to GD 18. Controls received 0.5% Tween-20 vehicle (1 ml/kg for rats and 10 ml/kg for mice). At parturition, newborns were observed for clinical signs and survival. All animals were born alive and initially appeared to be active. In the highest dosage groups (10 mg/kg for rat and 20 mg/kg for mouse), the neonates became pale, inactive, and moribund within 30-60 min, and all died soon afterward. In the 5 mg/kg (rat) and 15 mg/kg (mouse) dosage groups, the neonates also became moribund but survived for a longer period of time (8-12 h). Over 95% of these animals died within 24 h. Approximately 50% of offspring died at 3 mg/kg for rat and 10 mg/kg for mouse. Cross-fostering the PFOS-exposed rat neonates (5 mg/kg) to control nursing dams failed to improve survival. Serum concentrations of PFOS in newborn rats mirrored the maternal administered dosage and were similar to those in the maternal circulation at GD 21; PFOS levels in the surviving neonates declined in the ensuing days. Small but significant and persistent growth lags were detected in surviving rat and mouse pups exposed to PFOS prenatally, and slight delays in eye opening were noted. Significant increases in liver weight were observed in the PFOS-exposed mouse pups. Serum thyroxine levels were suppressed in the PFOS-treated rat pups, although triiodothyronine and thyroid-stimulating hormone [TSH] levels were not altered. Choline acetyltransferase activity (an enzyme that is sensitive to thyroid status) in the prefrontal cortex of rat pups exposed to PFOS prenatally was slightly reduced, but activity in the hippocampus was not affected. Development of learning, determined by T-maze delayed alternation in weanling rats, was not affected by PFOS exposure. These results indicate that in utero exposure to PFOS severely compromised postnatal survival of neonatal rats and mice, and caused delays in growth and development that were accompanied by hypothyroxinemia in the surviving rat pups. PMID- 12773773 TI - Exposure to perfluorooctane sulfonate during pregnancy in rat and mouse. I: maternal and prenatal evaluations. AB - The maternal and developmental toxicities of perfluorooctane sulfonate (PFOS, C8F17SO3-) were evaluated in the rat and mouse. PFOS is an environmentally persistent compound used as a surfactant and occurs as a degradation product of both perfluorooctane sulfonyl fluoride and substituted perfluorooctane sulfonamido components found in many commercial and consumer applications. Pregnant Sprague-Dawley rats were given 1, 2, 3, 5, or 10 mg/kg PFOS daily by gavage from gestational day (GD) 2 to GD 20; CD-1 mice were similarly treated with 1, 5, 10, 15, and 20 mg/kg PFOS from GD 1 to GD 17. Controls received 0.5% Tween-20 vehicle (1 ml/kg for rats and 10 ml/kg for mice). Maternal weight gain, food and water consumption, and serum chemistry were monitored. Rats were euthanized on GD 21 and mice on GD 18. PFOS levels in maternal serum and in maternal and fetal livers were determined. Maternal weight gains in both species were suppressed by PFOS in a dose-dependent manner, likely attributed to reduced food and water intake. Serum PFOS levels increased with dosage, and liver levels were approximately fourfold higher than serum. Serum thyroxine (T4) and triiodothyronine (T3) in the PFOS-treated rat dams were significantly reduced as early as one week after chemical exposure, although no feedback response of thyroid-stimulating hormone (TSH) was observed. A similar pattern of reduction in T4 was also seen in the pregnant mice. Maternal serum triglycerides were significantly reduced, particularly in the high-dose groups, although cholesterol levels were not affected. In the mouse dams, PFOS produced a marked enlargement of the liver at 10 mg/kg and higher dosages. In the rat fetuses, PFOS was detected in the liver but at levels nearly half of those in the maternal counterparts, regardless of administered doses. In both rodent species, PFOS did not alter the numbers of implantations or live fetuses at term, although small deficits in fetal weight were noted in the rat. A host of birth defects, including cleft palate, anasarca, ventricular septal defect, and enlargement of the right atrium, were seen in both rats and mice, primarily in the 10 and 20 mg/kg dosage groups, respectively. Our results demonstrate both maternal and developmental toxicity of PFOS in the rat and mouse. PMID- 12773774 TI - Ozone exposure enhances endotoxin-induced mucous cell metaplasia in rat pulmonary airways. AB - Coexposure to different airborne pollutants can be more toxic to airway epithelium than an inhalation exposure to a single pollutant. We have previously reported that coexposure to ozone, the primary oxidant gas in photochemical smog, and unique inflammatory biogenic substances such as allergens or bacterial endotoxin, results in augmented epithelial and inflammatory responses in rat nasal airways (M. V. Fanucchi et al., 1998, Toxicol. Appl. Pharmacol. 152, 1-9; J. G. Wagner et al., 2002a, Toxicol. Sci.67, 284-294). In the present study, we investigated the toxic interaction of ozone and endotoxin on the respiratory epithelium in the pulmonary airways of laboratory rodents. F344 rats were intranasally instilled with 0, 2, or 20 microg endotoxin dissolved in sterile saline (150 microl/nasal passage). Six h after instillation rats were exposed to air or 1 ppm ozone for 8 h. One day later, endotoxin and ozone exposures were repeated. Three days after the last exposure, rats were sacrificed, the lungs were lavaged with saline, and the collected bronchoalveolar lavage fluid (BALF) was analyzed for inflammatory cells and secreted mucosubstances (mucin 5AC). Lung tissues were processed for light microscopic examination and morphometric analysis of numeric density of epithelial cell populations and volume densities of intraepithelial mucosubstances (IM). Conducting airways were microdissected and analyzed by quantitative RT-PCR to determine steady-state mucin gene (rMuc5AC) mRNA levels in respiratory epithelium. Endotoxin instillation caused a dose-dependent increase in BALF neutrophils that was further increased twofold in ozone-exposed rats given 20 microg endotoxin. Mucin glycoprotein 5AC was elevated in BALF from rats exposed to 20 microg, but not 2 microg endotoxin. Exposure to ozone alone did not cause mucus hypersecretion, but ozone potentiated mucus secretion in rats given 2 or 20 microg endotoxin. Airways of rats exposed to air or ozone alone had scant amounts of IM. Endotoxin instillation induced a dose dependent increase in IM in airway epithelium that was significantly increased (twofold) in rats that were also exposed to ozone. Expression of rMuc5AC was induced in axial pulmonary airways by 2 and 20 microg endotoxin, and was increased further by ozone-exposure in rats instilled with 20 microg endotoxin. These data demonstrate that ozone exposure potentiates neutrophilic inflammation and mucus production and secretion elicited by a biogenic substance in rat pulmonary airways. PMID- 12773775 TI - Role of IL-1(beta) in endotoxin potentiation of deoxynivalenol-induced corticosterone response and leukocyte apoptosis in mice. AB - Endotoxin (lipopolysaccharide, LPS) and the trichothecenes are microbial toxins that are frequently encountered in food and the environment. Coexposure to LPS and the trichothecene deoxynivalenol (DON, vomitoxin) induces corticosterone dependent apoptosis in thymus, Peyer's patches, and bone marrow in mice. The purpose of this study was to test the hypothesis that interleukin-1beta (IL 1beta) plays a central role in corticosterone induction and subsequent leukocyte apoptosis in this model. Coexposure to LPS (0.1 mg/kg, ip) plus DON (12.5 mg/kg, po) was found to significantly upregulate splenic IL-1beta mRNA and IL-1beta protein expression in B6C3F1 mice, as compared to treatments with vehicle or either of the toxins alone. B6.129S7-IL1r1tm1Imx mice, which are functionally deficient for the IL-1 receptor 1, produced significantly less corticosterone upon coexposure to LPS plus DON than did corresponding wild-type (WT) C57BL/6J mice. Consistent with these findings, IL-1 receptor 1-deficient mice were recalcitrant to apoptosis induction in leukocytes as determined by assessment of DNA fragmentation assay and flow cytometry. Furthermore, intraperitoneal injection of IL-1 receptor antagonist (100 microgram/mouse, twice at 3 h intervals) in B6C3F1 mice significantly inhibited LPS plus DON-induced increases in plasma corticosterone, as well as apoptosis in thymus, Peyer's patches, and bone marrow. To confirm IL-1beta's capacity to induce apoptosis, B6C3F1 mice were injected with the cytokine (500 ng/mouse, ip) three times at 2 h intervals, and then corticosterone and apoptosis were monitored. Plasma corticosterone levels and thymus and Peyer's patch apoptosis in IL-1beta-injected mice were significantly higher at 12 h than in control mice. Plasma adrenocorticotropic hormone (ACTH) levels in LPS plus DON-treated B6C3F1 mice did not correlate with the induction of plasma corticosterone or leukocyte apoptosis. Taken together, the results indicate that IL-1beta is an important mediator of LPS plus DON induced corticosterone and subsequent leukocyte apoptosis and, furthermore, this cytokine possibly acts through an ACTH-independent mechanism. PMID- 12773776 TI - Comparing the effects of stage and duration of retinoic Acid exposure on amphibian limb development: chronic exposure results in mortality, not limb malformations. AB - Recently, high frequencies of malformations have been reported in amphibians across the United States. It has been suggested that the malformations may be the result of xenobiotic disruption of retinoid signaling pathways during embryogenesis and tadpole development. Therefore, a series of experiments were undertaken to examine life-stage specific effects of continuous retinoid exposure on Xenopus laevis. Continuous all-trans retinoic acid (RA) concentrations were delivered using a column saturator and a flow-through diluter system. Stage 8 embryos were exposed to RA concentrations ranging from 0.013 to 2 microgram/l. At the onset of hindlimb bud emergence (NF stage 48), a subset of tadpoles was moved to clean water, and remaining organisms were exposed continuously through metamorphosis. In addition, early limb-bud-stage tadpoles were exposed for 1 week, 2 weeks, or until tail resorption was complete, to eight concentrations of RA in the range of 0.031-3 microgram/l. RA exposure resulted in a concentration dependent increase in mortality and dysmorphogenesis in embryos at concentrations of 0.24 microgram/l and above. However, this early embryonic exposure did not result in hindlimb abnormalities in surviving tadpoles allowed to mature in clean water. RA did not induce limb malformations in any surviving tadpole exposed during larval stages. We are confident that the concentrations used were high enough, given that the highest concentration used resulted in 100% mortality within 2 weeks of initiating the exposure. This result suggests that other aspects of growth and development, which are not externally obvious, are more sensitive to retinoids than skeletal development. From these experiments and our previous work, we conclude that it is unlikely that retinoid mimics would produce the spectrum of limb malformations which recently have been observed in amphibians collected from the field. PMID- 12773777 TI - The effect on sperm production in adult Sprague-Dawley rats exposed by gavage to bisphenol A between postnatal days 91-97. AB - M. Sakaue et al. (2001,J. Occup. Health vol. 43, pp. 185-190) have described how oral exposure of sexually mature male rats to bisphenol A (BPA) between postnatal days (PND) 91-97 led to a reduction in daily sperm production (DSP) 5 weeks later (18 weeks of age). Activity was observed over the dose range 20 microgram/kg-200 mg/kg BPA, with an absence of activity over the dose range 2 ng/kg-2 microgram/kg BPA. There was no evidence of a dose response relationship over the active dose range (five orders of magnitude range). The observation of endocrine disruption (ED) effects for BPA at such low doses, and in sexually mature animals, was unexpected. It was therefore decided to mount an independent repeat of their study. A total of four independent studies were conducted according to the protocol used by Sakaue et al. Doses of 20 microgram/kg, 2 mg/kg, or 200 mg/kg BPA were administered to adult Sprague-Dawley (SD) rats over PND 91-97, and the studies were terminated when the rats reached the age of 18 weeks. Three different rodent diets were employed (RM3, Purina 5002, and CE2), the last of which had been used by Sakaue et al. BPA failed to give any evidence of ED activities, including the changes in DSP reported by Sakaue et al. 2001. During the course of these studies, the test protocol was adapted to coincide more precisely with that used by Sakaue et al.; this included restricting the number of animals per cage, removing bedding from the cages, and changing to the use of glass water bottles in the cages. The only thing of interest to emerge from our studies was the observation of a significant difference in DSP between the control groups of our first and second study. As the change in diet from RM3 to Purina 5002 was the major difference between those two studies, we conducted a repeat of the second study, but we were unable to confirm the differences seen between the first and second study. The probability that those differences arose either by chance, or as the result of intrinsic study-to-study variability, was strengthened by the absence of significant differences in the sperm parameters in a final (fifth) study where the sperm parameters for control animals maintained on the three different diets were compared under the conditions of the main experiments. No explanation for our failure to replicate the effects reported by Sakaue et al. is evident. A review of DSP values reported in the recent literature is provided and discussed, and it is concluded that use of the term DSP/g testis rather than DSP/testis could increase the sensitivity of DSP assessments. PMID- 12773778 TI - Pleural dosimetry and pathobiological responses in rats and hamsters exposed subchronically to MMVF 10a fiberglass. AB - Interspecies differences in pulmonary and pleural responses to the inhalation of natural mineral and synthetic vitreous fibers have been observed in chronic and subchronic studies. However, the reasons for these differences are not clearly understood. There are also fiber-specific differences in the outcome of chronic inhalation exposure to natural mineral and synthetic vitreous fibers. Whether these differences are dependent upon the ability of these fibers to translocate to the pleural space is unknown. The present study was conducted to compare retained fiber burdens and selected pathological responses in the pleural compartments of rats and hamsters following subchronic inhalation of MMVF 10a fiberglass, a fiber negative for tumorigenesis or fibrosis in chronic studies. Fischer 344 rats and Syrian golden hamsters were exposed for 4 or 12 weeks by nose-only inhalation at nominal aerosol mass concentrations of 45 mg/m3 (610 WHO fibers/cc). Pulmonary fiber burdens and pulmonary inflammatory responses were greater in rats than in hamsters. The total number of fibers in the lung was approximately three orders of magnitude greater than in the pleural compartment. Pleural burdens in the hamster (160 fibers/cm2 surface area) were significantly greater than burdens in similarly exposed rats (60 fibers/cm2 surface area) following 12 weeks of exposure. With time postexposure, pleural burdens decreased in hamsters but were essentially unchanged in rats. Pleural inflammatory responses in both species were minimal. In rats, pleural inflammation was characterized by increased numbers of macrophages and increases in mesothelial cell replication during the period of fiber exposure. In contrast, hamsters had increased numbers of macrophages and lymphocytes, and mesothelial-cell replication indices were elevated on the parietal pleura of the costal wall and diaphragm, with some of these responses persisting through 12 weeks of postexposure recovery. Taken together, the results suggest that differences among rodent species in pleural responses to inhaled fibers are due to a delivered dose of fibers and to the biological responses to the presence of the fibers. PMID- 12773779 TI - Innovative approaches to youth tobacco control. PMID- 12773780 TI - Innovative approaches to youth tobacco control: introduction and overview. PMID- 12773781 TI - Teen penalties for tobacco possession, use, and purchase: evidence and issues. AB - A handful of empirical studies have related changes in youth smoking to popular laws that penalise tobacco possession, use, and purchase (PUP). In this paper, we review the literature and outline reasons why PUP laws may be unlikely to reduce youth smoking significantly at the population level. In theoretical terms, we argue that PUP laws lack important features required for punishment to be effective in changing behaviour. In practical terms, PUP transgressions seem difficult to detect. Conceptually, there is potential for PUP laws to undermine conventional avenues of discipline, such as the parent-child relationship and the school environment. Strategically, PUP laws may divert policy attention from effective tobacco control strategies, relieve the tobacco industry of responsibility for its marketing practices, and reinforce the tobacco industry's espoused position that smoking is for adults only. To assist further debate and discussion, we identify research issues requiring attention. PMID- 12773782 TI - Regulatory strategies to reduce tobacco addiction in youth. AB - Preventing tobacco addiction and achieving cessation in established users are the cornerstones of efforts to reduce tobacco use and disease. It has been increasingly recognised that reducing tobacco toxin exposure has theoretical potential to reduce disease in continuing tobacco users. This has been controversial because such approaches also carry the potential to undermine prevention and cessation. As complicated as harm reduction issues are for adults, they are still more complicated for youth. Harm reduction is not a singular approach, but rather a concept that encompasses an extremely diverse array of potential approaches. These carry equally diverse potential risks and benefits. The regulatory framework (for example, whether or not the Food and Drug Administration regulates the approach) is also predicted to be a major factor in determining the consequences of harm reduction approaches. This paper examines the various issues and potential approaches concerning the application of harm reduction to youth. We conclude that although some carry great risk, others may actually support broader tobacco control efforts to prevent tobacco use and foster cessation in youth and adults. PMID- 12773783 TI - Teen smoking cessation. AB - Interest in adolescent smoking cessation has increased dramatically over the past several years, as researchers and practitioners have acknowledged the high rates of adolescents who smoke regularly and the low probability that adolescents who are regular smokers will stop on their own. The evidence base behind smoking cessation interventions for adolescents is also now starting to grow, but unfortunately the studies to date have frequently been plagued by major methodological problems. This paper summarises research conducted on adolescent smoking cessation, notes some of the methodological limitations of prior work, highlights approaches that show promise, discusses some of the challenges involved in addressing adolescent smoking cessation, and makes recommendations for future work. PMID- 12773784 TI - Youth tobacco prevention mass media campaigns: past, present, and future directions. AB - This paper focuses on countermarketing efforts aimed at curbing youth smoking. We review the literature on the effectiveness of tobacco countermarketing campaigns, characterise current state and national campaign approaches, present findings from qualitative approaches and laboratory experiments that explore a variety of messages (for example, health consequences, industry manipulation), and discuss newer, non-traditional approaches to countermarketing. In conclusion, we outline research needed to fill gaps in our existing knowledge and discuss future directions in tobacco countermarketing aimed at youth. PMID- 12773785 TI - The potential of the internet as a medium to encourage and discourage youth tobacco use. AB - The internet is fast becoming a new battleground between tobacco control advocates and pro-tobacco forces, and this new media will certainly have a greater impact on tobacco use behaviour in the future. This paper reviews how the internet can encourage youth smoking by providing youth access to tobacco products and offering content that glamorises smoking lifestyle and culture, particularly in hundreds of websites and chat rooms. These sites feature pictures of celebrity smokers, provide information about smoking in movies, and provide smoking advice to teen smokers. In contrast, youth smoking is discouraged on online grassroots advocacy and countermarketing websites. Although these strategies show promise, more research is needed to evaluate their impact. Recommendations are made for future research to study pro-smoking internet content and ways to counteract it, as well as to monitor the online activities of the tobacco companies. Finally, some of the challenges in addressing tobacco related internet content are discussed. PMID- 12773787 TI - Group discussions. PMID- 12773786 TI - Smoking on the rise among young adults: implications for research and policy. AB - This paper investigates hypotheses regarding the cause of the recent apparent increase in young adult smoking, compares trends in smoking among young adults with trends in the use of other substances, and considers the implications for youth tobacco control research and policy. Time series analyses of national data suggest that the recent observed increase in smoking among young adults is primarily an artefact of the almost simultaneous increase in smoking among high school students. In addition, however, it also appears that there have been real changes in smoking patterns among young adults. While many questions remain regarding recent trends in tobacco and other drug use among adolescents and young adults, what is known leads to a clarion call for increased intervention and policy action for the prevention and control of tobacco use among young adults in the USA. PMID- 12773789 TI - Chromosome pairing in tetraploid rye with monosomic-substitution wheat chromosomes. AB - In tetraploid rye with single-substitution wheat chromosomes - 1A, 2A, 5A, 6A, 7A, 3B, 5B, 7B - chromosome pairing was analysed at metaphase I in PMCs with the C-banding method. The frequency of univalents of chromosome 1A was considerably higher than that of the other four wheat chromosomes of genome A (6A, 5A, 7A and 2A). Among chromosomes of genome B, the lowest mean frequency of univalents was observed for chromosome 5B. In monosomic lines, wheat chromosomes 1A, 2A, 5A, 6A, 7A and 5B paired with rye homoeologues most often in rod bivalents and in chain quadrivalents (also including 3B). The 47% pairing of 5B-5R chromosomes indicate that the rye genomes block the suppressor Ph1 gene activity. In monosomic plants with chromosomes 5A, 2A, 6A, 7A and 5B, a low frequency of rye univalents was observed. It was also found that the wheat chromosomes influenced the pairing of rye genome chromosomes, as well as the frequency of ring and rod bivalents and tri- and quadrivalents. However, the highest number of terminal chiasmata per chromosome occurred in the presence of chromosomes 5A and 2A, and the lowest - in the presence of chromosomes 3B and 7B. In the presence of chromosome 5B, the highest frequency of bivalents was observed. The results of the present study show that the rye genome is closer related to the wheat genome A of than to genome B. The high pairing of wheat-rye chromosomes, which occurs in tetraploid rye with substitution wheat chromosomes, indicates that there is a high probability of incorporating wheat chromosome segments into rye chromosomes. PMID- 12773790 TI - The influence of rye cytoplasm on meiotic stability of tetraploid Secalotriticum. AB - Chromosome pairing in tetraploid Secalotriticum was analysed. In the studied plants wheat chromosomes in PMCs during metaphase I showed a higher degree of pairing, in comparison to the rye genome. This is reflected in a very low frequency of univalents and a higher frequency of ring bivalents. The occurrence of wheat univalents was dependent on wheat mixogenome. In plants with an unstabilized fourth homoeologous group, a heteromorphic bivalent 4A-4B was observed in 39.9% of PMCs, whereas in plants with an unstabilized seventh homoeologous group, chromosome 7A-7B pairing was found in all analysed cells. Rye univalents were present in all plants studied. The highest mean frequency of univalents and rod bivalents, both in wheat and in rye genomes, were recorded in plants whose first homoeologous group contained chromosome 1A. The mean number of terminal chiasmata per chromosome amounted to 1.78 in the wheat genome and 1.36 in the rye genome. It may be concluded that the plasmagenes in Secalotriticum did not increase the meiotic stability of the rye genome and also did not stabilize plant fertility. PMID- 12773791 TI - Leaf rust resistance genes of wheat: identification in cultivars and resistance sources. AB - Thirty-seven wheat cultivars originating from seven European countries were examined by using sequence tagged site (STS) markers for seven Lr (leaf rust = brown rust) resistance genes against the fungal pathogen of wheat Puccinia recondita f. sp. tritici (Lr9, Lr10, Lr19, Lr24, Lr26 and Lr37). Additionally, 22 accessions with various Lr genes from two germplasm collections were tested. A Scar (sequence-characterized amplified region) marker for Lr24 and a CAPS (Cleaved Amplified Polymorphic Sequence) marker for Lr47 were also used to identify those genes in the wheat accessions. Each marker amplified one specific DNA fragment. Three Lr gene markers were identified in wheat cultivars (Lr10, Lr26 and Lr37). Another four markers (Lr9, Lr19, Lr24 and Lr47) were found in breeding lines carrying leaf rust resistance genes. The results were compared with leaf rust resistance gene postulations made in previous studies, based on multipathotype testing. Markers for Lr10, Lr26 and Lr37 may be useful in marker assisted breeding. PMID- 12773792 TI - Determination of the ploidy level in chamomile (Chamomilla recutita (L.) Rausch.) strains rich in alpha-bisabolol. AB - The present study aimed to identify morphological traits whose performance depends on plant ploidy in Chamomilla recutita. Flowerhead diameter, 100 flowerhead weight, 1000-seed weight, pollen grain diameter, stoma length, and number of chloroplasts per guard cell, were examined in 5 tetraploid and 5 diploid strains. Out of these traits, stoma length, number of chloroplasts per guard cell, 100-flowerhead weight, and 1000-seed weight, proved to be significantly higher in tetraploid than in diploid strains, and can be used for indirect identification of tetraploid and diploid genotypes in various developmental stages of this species. PMID- 12773793 TI - Evaluation of genetic diversity and uniformity of head cabbage DH lines by the use of RAPD markers. AB - DH lines derived from cabbage cvs. Kamienna Glowa, Slawa z Enkhuizen and Langendijker, representing R1 generation, were analysed by the use of RAPD markers for their diversity and uniformity. For the evaluation of genetic diversity, eight primers yielding informative bands were used. Of the total of 83 RAPD bands scored in this study, 16.9% were polymorphic between a set of 13 DH lines. The similarity of the DH lines, estimated by Jaccard's coefficient, was depicted in the UPGMA dendrogram. Fourteen generated informative RAPD bands allowed the identification of DH lines developed from each cultivar. The evaluation of the uniformity for six closely related DH lines was possible by the use of three primers which generate one or two polymorphic bands. The lack of differences among ten plants of the five investigated DH lines manifested their uniformity. One line showed intraline polymorphism with two RAPD primers. The occurrence of the differences at the molecular level among ten plants indicated that their parental R0 plant was probably obtained from somatic cells, not by androgenesis. PMID- 12773794 TI - Transgenic rabbit producing human growth hormone in milk. AB - The gene construct WAP(6xHisThr):hGH containing the entire human growth hormone gene (hGH) under the rat whey acidic protein (WAP) promoter regulating the expression in mammary glands of mammals was prepared. The 5' end of the gene was modified by the addition of a sequence encoding six histidine residues and a sequence recognized by thrombin. The gene construct was introduced by microinjection into the male pronucleus of a fertilized oocyte. The founder male rabbit was obtained with the transgene mapping to chromosome 7. The presence of the growth hormone was confirmed in samples of milk collected during the lactation of F1 generation females. The growth hormone can be easily purified by affinity chromatography and cleavage by thrombin to an active form. PMID- 12773795 TI - Inbreeding effects on lifetime in David's deer (Elaphurus davidianus, Milne Edwards 1866) population. AB - The current population of David's deer is derived from 18 individuals kept in Woburn Abbey Park (England). The aim of this study is to evaluate the inbreeding rate as well as inbreeding depression in longevity. The recorded data have been extracted from the International Species Information System (ISIS). Complete records of 2042 individuals (born in 1947-2000) from zoological gardens were studied. The following four subsets of data were formed: all individuals, individuals over 31 days of life, sexually mature individuals (above 450 days old) and individuals with identified sex. Two models (including inbreeding coefficient as linear and quadratic covariables, respectively) have been employed. These computations were performed by the use of the DFREML package programs. This study has shown that average levels of inbreeding in the David's deer are relatively low (no exceed 0.028). The highest level of inbreeding was registered for mature individuals. In general, the average inbreeding on length of life was small for the above mentioned the inbreeding level (from 24 days to 77 days). However, on the basis estimated regression coefficients it can be suggested that an increase of inbreeding could lead to a drastic reduction of longevity. Relationships between inbreeding level and longevity are usually better described by quadratic partial regression (except for the oldest individuals). On the other hand, from a statistical point of view, a relatively low inbreeding level of the population studied is not suitable to derive the slope of the dependencies. PMID- 12773796 TI - Polymerase chain reaction-heteroduplex (PCR-HD) polymorphism within the bovine STAT5A gene. AB - Signal transducers and activators of transcription (STATs) are transcription factors mediating signals of various hormones and cytokines. STAT5A, previously known as the mammary gland factor (MGF), mediates the action of prolactin on milk protein gene expression in mammary epithelial cells. We used polymerase chain reaction-heteroduplex (PCR-HD) and sequencing methods for detection of nucleotide sequence polymorphism in intron 15 of the bovine STAT5A gene. A 281-bp gene fragment, from nt 12525 to nt 12806 (GenBank AJ 237937), was amplified with PCR, denatured and subjected to polyacrylamide gel electrophoresis to detect PCR-HD polymorphism. Three genotypes and two alleles were identified. DNA samples derived from homozygotes AA and BB were sequenced. A trinucleotide CCT deletion was found in the variant B of the STAT5A gene at position 12549. In a group of 72 beef bulls of various breeds and 49 Friesian bulls genotyped by PCR-HD mostly the AA genotype was found (from 83 to 61% depending on breed). The frequency of allele A varied between 0.91 and 0.77. Animals of genotype BB were found in Charolaise and Limousine breeds only. PMID- 12773797 TI - Prion octapeptide-repeat polymorphism in Polish Black-and-White cattle. AB - The study was carried out in a Polish Black-and-White cattle population, represented by 167 AI sires, 200 young tested bulls, 190 bull-dams, and 606 randomly chosen cows from commercial herds. The fragment of the bovine prion protein gene (PRNP) coding the octapeptide-repeat sequence, was identified by PCR analysis. Two different gene variants of 349 bp and 373 bp in size, produced three genotypes: PRNP 6/6, PRNP 6/5 and PRNP 5/5, respectively. Allele frequency in all examined populations, on average 0.894 for PRNP 6 and 0.106 for PRNP 5, shows a significant difference between the group of cows from commercial herds, characterised by high frequency of PRNP 5 (q = 0.137) in comparison to AI sires (q = 0.077), young tested bulls (q = 0.052) and bull-dams (q = 0.084). Moreover, both analysed female groups of bull-dams and cows from commercial herds are distinguished by the presence of PRNP 5/5 homozygous animals, which were not recorded in the AI sires and young tested bulls, and had never been recognised in earlier examined Holstein-Friesian populations. Analysis of the genetic equilibrium indicates a very high conformity between observed and expected number of animals in the separate PRNP genotype groups. However, some tendency of difference is observed in highly selected cows, qualified as bull-dams on the basis of very high level of milk performance traits. PMID- 12773798 TI - Genetic basis of autosomal dominant nocturnal frontal lobe epilepsy. AB - In this review the current literature regarding autosomal dominant nocturnal frontal lobe epilepsy (ADNFLE) is presented and discussed. This disease is caused by mutations of genes coding for subunits of neuronal acetylcholine receptor comprising the sodium/potassium ion channel. To date, three types of mutations of the gene encoding alpha4 subunit of acetylcholine receptor were described in multi-generation families in Australia, Spain, Norway and Japan. Two other types of mutations of the beta2 subunit were also reported in two families, one from Italy and the other from Scotland. Mutations were caused by substitutions of a single nucleotide or by several-nucleotide insertions and result in a decrease or an increase in the activity of the receptor, or its changes in the affinity to the ligand. Recent advances in molecular genetics have provided the means for a better understanding of human epileptogenesis at a molecular level, which facilitates clinical diagnosis and provides a more rational basis of therapy of this form of epilepsy. PMID- 12773799 TI - Prenatal detection of maternal UPD15 in a new case with i(15p) by Timing Replication Test (TRT) and methylation analysis. AB - DNA replication kinetics of the Prader-Willi/Angelman Critical Region (PWACR) was studied with and without synchronisation in human amniotic cell cultures obtained from 20 cases with normal karyotype and 4 cases with a marker of chromosome 15, respectively. A Timing Replication Test (TRT) was performed by synchronisation of amniotic cell cultures and followed by interphase FISH to analyse and compare the early/late replication patterns in SNRPN and UBE3A genes between the homologues of chromosome 15. Asynchronous replication patterns of the analysed genes were observed in both amniotic cell cultures but the percentage of interphase nuclei presenting with asynchronous replication was significantly increased in the cultures with synchronisation (40-51%), as compared to those without synchronisation (20-23%). The evaluations, performed by means of TRT, showed asynchronous replication patterns on control values: between 39% and 46% of cells in all the cases with inv dup(15). In contrast, the percentage of cells with asynchronous replication in the case with i(15p) was significantly decreased (3 6%), as compared to the control value, and it may be indicated by uniparental disomy of chromosome 15 (UPD15). In addition, those results have been confirmed by molecular evaluation, using the methylation diagnostic test for diagnosis of the Prader-Willi Syndrome. PMID- 12773800 TI - Analysis of polymorphism of three human Y-chromosome STR loci: DYS390, DYS392 and DYS393 in the population of northern Poland. AB - The study aimed at development of a multiplex PCR system for amplification of three Y-chromosome STR loci: DYS390, DYS392 and DYS393, and its application in haplotype polymorphism analysis in the population of northern Poland. Due to interactions between originally published primers, a new DYS392 primer pair was proposed. In a population of 158 unrelated males, 28 different haplotypes could be observed, 12 of which were seen only once. The haplotype diversity is 0.805. Distribution of haplotypes of the studied loci is specific to the population of northern Poland and distinguishes it from compared West-European populations. To our knowledge, this is the first report on a Y-STR multiplex system that can be analysed on native polyacrylamide gels. PMID- 12773801 TI - Inv(10) in a patient with hypogonadotropic hypogonadism. AB - Hypogonadotropic hypogonadism (HH) was diagnosed in a 22-year-old patient with 46,XY,inv(10) karyotype. It may be associated with some gene mutations of chromosome X, (KAL-1: Kallman syndrome; and DAX-1: congenital adrenal hypoplasia), as well as of certain autosomes, including chromosome 10. This study aimed to: (1) elucidate the aetiopathogenesis of the disease in the studied case: (2) diagnose chromosome aberrations as accurately as possible: and (3) determine if the observed clinical picture can be referred to the diagnosed chromosomal aberration or it is a mere coincidence. The FISH technique, with the use of non commercial DNA probes, was applied for a precise description of chromosome breaking points. The application of FISH enabled karyotype description: 46,XY, inv(10)(p15.2q11.22).ish inv(10)(p15.2q21.3)(p15x3)(q21x3)(p15conq21x2). The SSCP method revealed no mutation within the DAX-1 gene and no deletion in the KAL-1 gene. PMID- 12773802 TI - Retinoids: fascinating up-and-coming scenario. AB - Retinoids have been in sharp focus ever since their introduction 30 years ago. They include any drug (s) that bind to retinoid receptors and elicit a biological response. Enormous information on the subject seems to embroil the recent literature. Practically it is impossible to clearly comprehend the undercurrents. The meticulously dispensing text envisages surmounting the perspective reader's predicaments. Accordingly, retinoids and their related facets namely retinoid receptors, classification, mode of action, and the pharmacological diversity have been precisely defined. Commonly used systemic retinoids too have been given a substantial fresh look along with their monitoring. Overall, adverse effects and relative and absolute contraindications have been scrupulously incorporated. Human immuno deficiency virus (HIV) and isoretinoid for acne, in particular, have been highlighted. Micronized isotretinoin formulations have also been taken care so also commonly used topical retinoids. Tretinoin and their newer formulation have also been accounted for along with tretinoin polymer cream. Adapalene, a new chemical entity possessing a unique physico-chemical activity similar to that of tretinoin has also been dealt with. Newer retinoids are likely to be a subject of intrigue. A focus on future potentials of retinoids is its special ingredient. The inclusion of details of rexinoid the most recent introduction in their purview is likely to invoke interest to further consolidate its reckoning in future. All in all the text of the paper should provide an insight into the current rumbling around retinoids. PMID- 12773803 TI - Phakomatosis pigmentovascularis II A and II B: clinical findings in 24 patients. AB - Nearly 200 cases of phakomatosis pigmentovascularis (PPV) have been reported worldwide, most of them of Japanese origin. There are 5 types and 10 subtypes of PPV. Its etiology might be explained by the twin spotting phenomenon. The relative frequency of PPV at the National Institute of Pediatrics was 5.8 per 100,000 pediatric patients and 0.634 per 100,000 dermatological patients. We report 24 cases of PPV with an average follow up of 5 years and the following findings: PPV type II A in 4 male and 2 female patients with melanosis bulbi in 3 and glaucoma in 1. PPV type II B in 7 male and 11 female patients, with melanosis bulbi in 9, glaucoma in 9, iris mammillations in 2, Sturge Weber syndrome in 6 female patients, and Klippel-Trenaunay syndrome in 2 males, hemifacial, hemicorporal, or limb hypertrophy without venous insufficiency in 6 female and 4 male patients. During the follow-up time of 60 months, progressive fading of melanotic and vascular macules were observed in 7 patients. No other types of PPV were found. Systemic involvement in PPV was related to the body surface area affected by the vascular macules. Ectodermal and mesodermal migration disorders might be involved in the pathogenesis of PPV. PMID- 12773804 TI - A cardiac allograft recipient with Bowen's disease on a finger and concurrent perianal bowenoid papulosis. AB - We report a patient who developed Bowen's disease of the finger and bowenoid papulosis of the perianal area after cardiac transplantation. Human papillomavirus (HPV) type 16 only, not any skin-related or epidermodysplasia verruciformis-related types, was detected in both lesions by in situ hybridization and polymerase chain reaction. The same virus type was identified in both the tumor of the finger and the perianal area, which suggests contact transmission. HPV 16 has often been associated with malignant changes and may be at least one source of the malignancies that are more common in immunosuppressed patients. The presence of a potentially oncogenic type of the HPV in an immunosuppressed patient highlights the importance of regular follow-up of such patients. PMID- 12773805 TI - Inguinal keratotic Basal cell carcinoma mimicking giant solitary trichoepithelioma. AB - Keratotic basal cell carcinoma may not only clinically but also histologically share more or less the same features with giant solitary trichoepithelioma. It can be difficult to distinguish these two entities from each other, even for an experienced dermatopathologist. We present an unusual case of inguinal keratotic basal cell carcinoma mimicking giant solitary trichoepithelioma in a 56-year-old woman with a finger-like tumor of 20 years duration. The patient presented with an asymptomatic, skin colored, firm, nonulcerative, nodular lesion. Scanty mitotic activity and apoptotic cells were the histopathologic findings against basal cell carcinoma, whereas absence of papillary mesenchymal bodies, presence of peritumoral lacunae detected only around the solid areas, and accumulation of amyloid-like hyalinized material were the findings in favor of basal cell carcinoma. This case illustrates that keratotic basal cell carcinoma must be taken into account in the differential diagnosis of inguinally located solitary, polypoid masses, especially giant solitary trichoepithelioma. PMID- 12773806 TI - Hydroa vacciniforme-like eruptions in a patient with chronic active EB virus infection. AB - We report a case of chronic active Epstein-Barr (EB) virus infection (CAEBV) associated with skin eruptions mimicking hydroa vacciniforme (HV) in a 4-year-old boy. The patient had repeated episodes of vesiculo-necrotic eruptions on the face, scalp, and bilateral forearms one year before the first visit to our department. General symptoms including fever, hepatosplenomegaly, abnormal liver function, and cervical lymph node swelling were noted three months before the first visit. At the first visit, small, bean-sized, erythemic papules with central necrosis were observed on the face and anterior chest wall. Thumb-sized ulcers with crust were present on the bilateral forearms. Histopathological examination of an erythematous lesion in the submandibular area revealed parakeratosis with a thick crust, mild spongiosis in the epidermis, and a dense infiltration of lymphoid cells into the dermis and perivascular space. Laboratory examination showed EBNA x 40, EBV VCA IgG x 1,280, and EBV DNA (PCR) 8 x 10(4). EBV-encoded small nuclear RNA (EBER) positive cells were detected in the dermis by an in situ hybridization (ISH) method. Large granular lymphocytes (65%) with the NK cell phenotype were found in the peripheral blood. A real time PCR method showed 171,741 copies/ micro g DNA in CD 16 positive cells. Although latent EBV infection-associated eruptions have been documented, detailed skin manifestations in CAEBV are less well known. PMID- 12773807 TI - Disseminated cellulitic cryptococcosis in the setting of prednisone monotherapy for pemphigus vulgaris. AB - Cryptococcal infections are seen mainly in immunocompromised hosts. The cutaneous manifestations of infection can include papules, plaques, nodules, vesicles, ulcers, ecchymosis, and, rarely, cellulitis. Cryptococcal meningitis is a life threatening complication of this disease. Pemphigus vulgaris is a rare auto immune blistering disease that can also be life-threatening. Treatment of pemphigus commonly entails both corticosteroids and steroid-sparing agents. We present a case of disseminated cryptococcal infection in a patient with pemphigus vulgaris treated with high dose corticosteroids as monotherapy. This case provides an opportunity to discuss the difficulty of managing two potentially mortal conditions in which the treatment of pemphigus vulgaris may exacerbate the disseminated cryptococcal infection. PMID- 12773808 TI - Oral ivermectin treatment in two cases of scabies: effective in crusted scabies induced by corticosteroid but ineffective in nail scabies. AB - We report two cases of scabies treated with oral ivermectin (200 micro g/kg). Case 1, a 72-year-old man, developed crusted scabies with the use of oral corticosteroids due to a misdiagnosis by an earlier physician. The patient was successfully treated with two doses of oral ivermectin at a 7 day interval with concomitant topical use of crotamiton and keratolytic agents. However, the nail scabies in this patient failed to respond to these treatments. Live mites were detected from all his toenails two weeks after the second dose of ivermectin. A complete cure of the nail scabies was achieved by occlusive dressing of 1% gamma BHC on all toenails for one month. Case 2, a 52-year-old woman, had been treated with oral corticosteroid for mesangial nephritis. She developed common scabies, but a topical scabicide, crotamiton, was not effective. Two weeks after treatment with a single dose of oral ivermectin, eggs were still detected from a burrow on her trunk. Her treatment was completed after a further two doses of oral ivermectin were administered at 7 day intervals. In both patients, the administration of oral ivermectin did not induce any clinical or laboratory side effects. Oral ivermectin is effective for crusted scabies, but not effective for nail scabies. Two doses of oral ivermectin, administered with a one-week interval, is an appropriate treatment regimen. PMID- 12773809 TI - Unilesional mycosis fungoides mimicking Bowen's disease. AB - Cutaneous T-cell lymphoma includes a heterogeneous group of lymphomas that share the unique feature of T lymphocytes which are tropic to the skin. Clinically, the presentation of this disease is protean and can range from a single lesion to generalized erythroderma. Unilesional mycosis fungoides, a rare variant, is presented here. Its morphology mimicked squamous cell carcinoma in situ, and was refractory to conventional therapies. A review of the treatment of this subset is also presented. PMID- 12773810 TI - Porokeratosis of mibelli with nail dystrophy. PMID- 12773811 TI - A case of median rhomboid glossitis. PMID- 12773812 TI - The Saccharomyces cerevisiae bud-neck proteins Kcc4 and Gin4 have distinct but partially-overlapping cellular functions. AB - In the budding yeast S. cerevisiae, Swe1 delays the onset of mitosis by phosphorylation and inactivation of the cyclin-dependent kinase Cdc28, thereby relaying the morphogenetic signal to the cell cycle. Hsl1/Nik1, Kcc4 and Gin4 are structurally homologous protein kinases that localize to the bud neck and negatively regulate Swe1 by phosphorylation. We report here that Kcc4 and Gin4 have partially overlapping but essentially distinct cellular functions. Deletion of KCC4 had a similar effect to GIN4 deletion, causing moderate defects in bud formation at stationary phase; overexpression of Kcc4 inhibited cell growth. KCC4 showed functional interaction with GIN4 in cdc28 mutants, and both Kcc4 and Gin4 proteins physically interacted with Swe1 in vitro. However, unlike gin4delta cells, kcc4Delta cells were not elongated but multi-budded at stationary phase, and showed resistance to 0.04% SDS and 0.003% calcofluor white. In light of the observation that Kcc4 and Gin4 specifically associate with distinct septin proteins, we propose that the observed functional distinction between Kcc4 and Gin4 is due to differences in septin association partners. PMID- 12773813 TI - Cloning and characterization of two full-length cDNAs, TaGA1 and TaGA2, encoding G-protein alpha subunits expressed differentially in wheat genome. AB - In the present study, we identified and characterized two cDNAs, named TaGA1 and TaGA2, encoding alpha subunits of heterotrimeric G proteins synthesized from one week-old seedling mRNAs of common wheat cv. S615 using RACE PCR and RT-PCR methods. The clone TaGA1 contained an open reading frame that encoded a protein consisting of 383 amino acid residues with a molecular mass of 51.3 kDa, whereas the clone TaGA2 contained an open reading frame encoding 390 amino acids with a molecular mass of 52.5 kDa. At the amino acid level, both cDNAs (TaGA1 and TaGA2) showed 70-96% and 30-40% homologies to plant and animal G-protein alpha (G alpha) subunits, respectively, and 97.7% homology to each other. The regions essential for binding to GTP were conserved among all G alpha subunits in higher plants and mammals examined. However, the C-terminal amino acid sequences of TaGA1 and TaGA2 were similar to those of cereal G alpha subunits (rice and barley) but were different from the analogous sequences of mammalian G alpha subunits as well as from those of the leguminous and Solanaeceous G alpha subunits. Southern analysis revealed that the hexaploid wheat genome contained three major copies of G alpha subunit gene with a few less homologous copies. The analysis of the expression for G alpha subunit genes in wheat showed that both TaGA1 and TaGA2 mRNAs were abundant in one-week-old seedlings, immature seeds harvested one-week after anthesis, young spikes and internodes, indicating constitutive expression patterns in all of the organs tested. Especially, young spikes and internodes exhibited increased levels of mRNA accumulation, suggesting that G alpha subunit gene is highly expressed in actively elongating and fast growing tissues. Moreover, both TaGA1 and TaGA2 showed genome-specific expressions in wheat and may participate in the light-regulated growth and development of the seedlings. PMID- 12773814 TI - Topological incongruence between nuclear and chloroplast DNA trees suggesting hybridization in the urophyllum group of the genus Fagopyrum (Polygonaceae). AB - We performed phylogenetic analyses of Fagopyrum species in the urophyllum group based on nucleotide sequences of two nuclear genes, FLORICAULA/LEAFY (FLO/LFY) and AGAMOUS (AG), and three segments of chloroplast DNA (cpDNA), rbcL-accD, trnK intron, and trnC-rpoB spacer. The FLO/LFY and AG sequences turned out to be phylogenetically more informative at the intrageneric level than the cpDNA sequences. Congruence among these gene trees, inferred by a maximum-likelihood (ML) method, demonstrated that topologies were partially incongruent between the nuclear and chloroplast DNA phylogenies. The nuclear DNA sequence data supported a monophyletic relation of F. statice, F. gilesii, and F. jinshaense, whereas the former two species formed another monophyletic relation with the F. capillatum-F. gracilipes-F. gracilipedoides-F. rubifolium clade excluding F. jinshaense in the synthetic cpDNA phylogeny. In addition, two divergent sequences of FLO/LFY were found in F. rubifolium (tetraploid). One of these was sister to F. gracilipedoides and another was sister to F. statice, and a monophyletic relation of these two genes was rejected by a bootstrap analysis. These results suggest that hybridization may have occurred during diversification of Fagopyrum species in the urophyllum group, and that F. rubifolium is possibly allotetraploid species. PMID- 12773815 TI - Development of a quantitative pachytene chromosome map in Oryza sativa by imaging methods. AB - A higher GC content region of an Oryza sativa chromosome can be specifically visualized by double staining with propidium iodide (PI) and 4, 6-diamidino-2 phenylindole (DAPI). This procedure allows identification of chromosome 9 from the other rice chromosomes at the pachytene stage. Using rice chromosome 9 as a model, an imaging method to construct a pachytene chromosomal map was developed by quantifying the fluorescence profile (FP) of each chromomere. The pachytene map of chromosome 9 consists of twenty-two chromomeres including four chromomeres within the nucleolar organizing region (NOR) and satellite region. The pachytene map was compared with the corresponding somatic prometaphase map and the linkage map. The differences among the three maps indicate that each map depicts specific biological information, which is difficult to be substituted by the other maps. PMID- 12773816 TI - Genetic studies on premating isolation in Drosophila simulans. II. A D. simulans line highly crossable to males of D. yakuba and D. teissieri. AB - Females of the S1 line of Drosophila simulans, an isofemale line from a natural population at Mishima, Japan, were found to show high crossability to males of D. yakuba and D. teissieri. With D. yakuba males, 24.0-58.5% of the S1 females were inseminated, while the rate was only 0.0-6.3% for the control lines. The high crossability was ascribed to the genes on the X and third chromosomes. With D. teissieri males, 38.0-51.5% of the S1 females were inseminated, while the rate was only 0.0-6.2% for the control lines. The crossability was ascribed to the genes on the third chromosome, and possibly the second chromosome genes acted additively. The crossabilities of hybrid females between the S1 and the control lines were intermediate between the parental lines both to D. yakuba males and to D. teissieri males. PMID- 12773817 TI - cDNA cloning of the chicken DDB1 gene encoding the p127 subunit of damaged DNA binding protein. AB - DDB (damaged DNA-binding protein) is a heterodimer, comprised of p48 (DDB2) and p127 (DDB1) subunits, which has a high affinity for a variety of DNA lesions including UV-photoproducts. The mutations in DDB2 gene have been found in a subset of xeroderma pigmentosum complementation group E patients. However, no natural mutation has been identified so far in the cDNA of human DDB1 and the precise roles of DDB1 are still unknown. We have cloned the DDB1 cDNA from the chicken B lymphocyte line DT40 and revealed an open reading frame of 3420 bp encoding a polypeptide of 1140 amino acids, which is identical in size to the orthologs of human, monkey, mouse, rat and Drosophila melanogaster in databases. The amino acid sequence deduced from the chicken DDB1 cDNA shows a high homology to the mammalian DDB1 orthologs (96-97% identity). Northern blot analysis using 5' portion of the chicken DDB1 cDNA as a probe detected a single transcript of ~ 4.3 kb in chicken DT40 cells as well as in human HeLa cells and mouse embryonic fibroblasts. Furthermore, the chicken DDB1 (tagged with enhanced GFP) transiently expressed in human cells mainly localized in the cytoplasm, and coexpression of human DDB2 dramatically changed the localization from the cytoplasm to nucleus. These results suggest that DDB1 is evolutionarily conserved in the primary structure and function, and may play a fundamental role in higher eukaryotes. PMID- 12773818 TI - Molecular phylogeny of Japanese Rhinolophidae based on variations in the complete sequence of the mitochondrial cytochrome b gene. AB - Microchiroptera have diversified into many species whose size and the shapes of the complicated ear and nose have been adapted to their echolocation abilities. Their speciation processes, and intra- and interspecies relationships are still under discussion. Here we report on the geographical variation of Japanese Rhinolophus ferrumequinum and R. cornutus using the complete sequence of the mitochondrial cytochrome b gene to clarify the phylogenetic positions of the 2 species as well as that of Rhinolophidae within the Microchiroptera. We have found that sequence divergence values within each of the 2 species are unexpectedly low (0.07%-0.94%). We have also found that there is no local specificity of their mtCytb alleles. On the other hand, the divergence values for Japanese Microchiroptera (12.7%-16.6%) are much higher than those for other mammalian genera. Similarly, the values among five genera of Vespertilionidae were 20.5%-27.3%. Phylogenetic analysis shows that the 2 species of family Rhinolophidae in the suborder Microchiroptera belong to the Megachiroptera cluster in the constructed maximum parsimony tree. These results suggest that the speciation of Rhinolophidae involved its divergence as an independent lineage from other Microchiroptera, and other microbats might be paraphyletic. In addition, the tree also shows that the order Chiroptera is monophylitic, and the closest group to Chiroptera is the ungulates. PMID- 12773819 TI - Characterization of the S-RNase promoters from sweet cherry (Prunus avium L.). AB - Genomic DNA fragments containing the S(3)-, S(4)-, and S(6)-RNase genes were isolated from the sweet cherry (Prunus avium L.) and sequenced. Comparison of the 5'-flanking sequences of these three S-RNases indicated that a highly conserved region (designated CR) existed just upstream from the putative TATA boxes. We postulate that CR contains cis-regulatory element(s) involved in pistil expression. To examine the activity of the isolated S-RNase promoters of sweet cherry in the pistil, we transiently introduced approximately 650-bp fragments of the S(4)- and S(6)-RNase promoters fused to beta-glucuronidase (GUS) gene into the pistil of the petunia using a particle bombardment technique. Histochemical analysis showed that the 5'-flanking region of each S-RNase was active in the pistil. This suggests that cis-regulatory element(s) for pistil-specific expression may exist(s) within the 650-bp region upstream from the TATA box in the sweet cherry S-RNase promoter. PMID- 12773820 TI - Presence of isochore structures in reptile genomes suggested by the relationship between GC contents of intron regions and those of coding regions. AB - Vertebrate genomes are mosaics of isochores. On the assumption that marked differences exist in the isochore structure between warm-blooded and cold-blooded animals, variations among vertebrates were previously attributed to adaptation to homeothermy. However, based on the data of coding regions from representatives of extant vertebrates, including a turtle, a crocodile (Archosauromorpha) and a few kinds of snakes (Lepidosauromorpha), it was recently hypothesized that the common ancestors of mammals, birds and extant reptiles already had the "warm-blooded" isochore structure. To test this hypothesis, the nucleotide sequences of alpha globin genes including non-coding regions (introns) from two snakes, N. kaouthia and E. climacophora, were determined (accession number: AB104824, AB104825). The correlation between the GC contents in the introns and exons of alpha-globin genes from snakes and those from other vertebrates supports the above hypothesis. Similar analysis using data for exons and introns of other genes obtained from the GenBank (Release 131) also support the above hypothesis. PMID- 12773821 TI - The cost of VA care: lessons of the evaluating VA costs study. Preface. PMID- 12773822 TI - Estimating private sector values for VA health care: an overview. AB - OBJECTIVES: To provide an overview of methods used to establish what taxpayer costs would be if all Veterans Health Administration (VA) patient care were paid for by the federal government but provided in the private sector. METHODS: Study assumptions included (1) that there would be a hypothetical policy change to pay for VA care through a Medicare-based fee-for-service program, (2) that the VA coverage benefit would not change, (3) that practice styles would remain the same, and (4) that there would be no impact on market values. To achieve the objective, project staff adapted Medicare payment schedules and guidelines, where available, with oversight of an advisory committee with VA and non-VA expertise in costs and data. For six sites, detailed payments were estimated using VA utilization databases and software and Medicare rate schedules available in the private sector. Overhead, interest on capital, and malpractice costs were added to VA-reported operating costs. Patient severity was examined, and patient-level costs were explored. FINDINGS: Detailed methods for pricing seven types of health services are presented. Three methods articles focus on process issues. DISCUSSION: Because VA care is not directly comparable with private sector health care as a result in part of differences in benefits covered and the scope of services provided, estimating costs for this care based on a private sector model requires careful consideration of market valuation approaches. The articles in this supplement describe the methods used to estimate market values for VA care so that other researchers can use them in future studies. PMID- 12773823 TI - Methods for estimating private sector payments for VA acute inpatient stays. AB - OBJECTIVES: To describe methods for estimating hypothetical private sector payments for Veterans Health Administration (VA) acute inpatient stays. METHODS: We assumed all VA hospitalizations would have occurred under a hypothetical VA system that paid private sector providers but had the current benefit package for VA patients. We compared aggregate budgets for VA inpatient care (less physician salaries) at six VA hospitals over federal fiscal year 1999 to aggregated hypothetical private sector payments developed using VA diagnosis-related groups matched to metropolitan-based average Medicare payments. Counts of care came from the VA's statistical analysis system (SAS) inpatient files. Inpatient stays with both medical or surgical and psychiatric or rehabilitation care were counted as two stays. An external auditor conducted three reviews of VA coding practices during the study year, and the appropriateness of admissions was examined using a commercial utilization review tool. RESULTS: For 30,518 inpatient discharges, hypothetical payments were $188 million, compared with the VA budget of $171 million. Fifteen of the 25 most frequent diagnosis-related groups in the VA were also in the top 25 for Medicare in 1998 and 1999. Audits established that the overall financial impact of VA coding problems was similar to that in the private sector. DISCUSSION: Differences in organization, practice, and incentives limit estimates of the financial impact of shifting VA acute inpatient care to the private sector. PMID- 12773824 TI - Estimating private sector professional fees for VA providers. AB - OBJECTIVES: To describe new methods used to estimate inpatient and outpatient Medicare-based professional fees for Veterans Health Administration (VA) services. METHODS: National VA utilization files provided estimates of inpatient physician services, whereas local provider and utilization files gave counts of outpatient services by physicians, nurse practitioners, physician assistant, clinical psychologists, and clinical social workers. Services from ambulatory surgery, emergency room, and clinics (eg, dermatology and gastroenterology) were coded by study health information management staff (coders). VA-based billing information was edited against Medicare guidelines. Estimates for VA services without comparable Medicare fees were obtained from other commercial sources. RESULTS: Hypothetical professional fees for VA services were 17% more ($109 million vs. $93 million) than the VA budget for physicians over 1 fiscal year at six sites. Total payments of nearly $21 million were generated for VA inpatient care. In fiscal year 1999, there were 30,209 admissions (of which 4549 were psychiatric) to the study sites; 30,518 discharges; 229,783 inpatient days, including 27,235 in critical care units; and 38,348 surgical days of care. DISCUSSION: Differences between the VA and the private sector maybe overstated because VA salaries of nonphysicians were not included in the VA budgets. Conversely, the extent to which VA professional services were undercounted in VA information systems used in this study may understate the difference. Future research may consider additional data collection approaches or information systems enhancements to enumerate more accurately all provider services that are reimbursable in the private sector. PMID- 12773825 TI - Methods to determine private sector payment for VA outpatient services: institutional payments to providers. AB - OBJECTIVES: To describe methods used to estimate hospital institutional (facility) payments for providing Veterans Health Administration (VA) outpatient services. METHODS: A series of audits compared the accuracy of outpatient coding at six VA medical centers in federal fiscal year 1999 with private sector standards. Outpatient records were processed through industry standard software to determine validity and remove inappropriate services. Private sector payments were estimated by applying average payment data from Medicare cost reports and Medicare outpatient prospective payment schedules to counts of VA services. RESULTS: Coding audits found little difference in accuracy between VA and the community. Physician visits generated the most estimated payments and deviated most from Medicare payment experience. Radiology and laboratory services were the next highest expenditure categories for both the VA and Medicare. The proportion of radiology payments in VA data was notably lower and ambulatory surgery notably higher than Medicare's experience. Within major categories, the relative rankings of VA and Medicare services were consistent. DISCUSSION: Differences in payment criteria make exact cost comparisons of hospital-based and office-based settings difficult, particularly physician visits. Two VA clinical software applications, radiology and laboratory, provide information not readily convertible to a claims format; these applications need significant changes to be used for these purposes. They understate radiology services and overstate laboratory services compared with private sector standards. In addition, the laboratory application contains inappropriate or unspecified codes that cannot be accurately valued for many reasons. PMID- 12773826 TI - Estimating private sector payments for VA specialized inpatient care. AB - OBJECTIVES: To describe methods for estimating what payments to private sector providers might be for specialized inpatient care in the absence of Veterans Health Administration (VA) facilities. METHODS: Psychiatric, rehabilitation, domiciliary, partial and day hospitalization, and psychiatric residential treatment programs at six study sites were audited for program content that would meet Medicare criteria for excluding providers from the prospective payment system. A 10% sample of patients in each program was also audited to see if they met VA program criteria. For programs similar to prospective payment system exempt community-based providers, total days of care were valued at per diem rates calculated for those Medicare providers. RESULTS: Not all specialized programs at the study sites were similar to private sector programs. Day hospitalization programs did not involve physicians, and inpatient psychiatric care was judged to be payable under acute diagnosis-related groups. Blind rehabilitation was different from any private sector program identified. For programs qualifying under Medicare exclusion rules, a majority of patients would meet criteria with minor changes in VA documentation. DISCUSSION: Researchers need to separate specialized inpatient care from acute services in estimating payments. This caution applies especially to rehabilitation, psychiatric, and long-term care, often provided to patients in VA acute bedsections. As with Medicare, benefits extend to higher costs of care not correlated with current measures of acuity or diagnoses. Medicare bases payments on the costs of the specialized providers of this care. PMID- 12773827 TI - What would VA nursing home care cost? Methods for estimating private sector payments. AB - OBJECTIVES: To describe the methods used to validate Veterans Health Administration (VA) nursing home acuity data and estimate hypothetical payments for nursing home patients in VA-based and community-based units. METHODS: For a sample of VA-based and community-based nursing home patients at six sites, auditors validated the resource utilization classifications from the most recent complete full or quarterly assessments. Scores were averaged to obtain an acuity index for each nursing home population. Per diem rates were calculated for a fully phased-in Medicare prospective payment system, a transitional prospective payment system for free-standing and hospital-based nursing homes, and average national Medicaid benefits based on VA patients in community nursing facilities. Days of care came from each site's end of year gains and losses financial statement. Nursing home estimates were calculated by multiplying together the number of days of care, the per diem, and the acuity index. RESULTS: The VA acuity information was valid. Generally, veterans' dependencies and depression were underscored (similar to the practice for non-VA patients). The cost of patients' nursing home care absent VA facilities would depend on the types of nursing homes in which they were placed. The most costly option (hospital-based facilities with cost exemptions) would cost 3.5 times the least costly. Only the Medicaid-only estimate was lower than actual VA expenditures. DISCUSSION: Future research on nursing homes must relate quality to the cost of care to help policy makers assess the value of different options for providing that care. PMID- 12773828 TI - Methods for estimating and comparing VA outpatient drug benefits with the private sector. AB - OBJECTIVES: To estimate and compare Veterans Health Administration (VA) expenditures for outpatient pharmaceuticals for veterans at six VA facilities with hypothetical private sector costs. METHODS: Using the VA Pharmacy Benefits Management Strategic Health Care Group (PBM) database, we extracted data for all dispensed outpatient prescriptions from the six study sites over federal fiscal year 1999. After extensive data validation, we converted prescriptions to the same units and merged relevant VA pricing information by National Drug Code to Redbook listed average wholesale price and the Medicaid maximal allowable charge, where available. We added total VA drug expenditures to personnel cost from the pharmacy portion of that medical center's cost distribution report. RESULTS: Hypothetical private sector payments were $200.8 million compared with an aggregate VA budget of $118.8 million. Using National Drug Code numbers, 97% of all items dispensed from the six facilities were matched to private sector price data. Nonmatched pharmaceuticals were largely generic over-the-counter pain relievers and commodities like alcohol swabs. The most commonly prescribed medications reflect the diseases and complaints of an older male population: pain, cardiovascular problems, diabetes, and depression or other psychiatric disorders. CONCLUSIONS: Use of the VA PBM database permits researchers to merge expenditure and prescription data to patient diagnoses and sentinel events. A critical element in its use is creating similar units among the systems. Such data sets permit a deeper view of the variability in drug expenditures, an important sector of health care whose inflation has been disproportionate to that of the economy and even health care. PMID- 12773829 TI - Methods to estimate and compare VA expenditures for assistive devices to Medicare payments. AB - OBJECTIVE: To describe the methods used to estimate and compare Veterans Health Administration (VA) annual expenditures for assistive devices and their repair at six VA hospitals with payments for those same devices in the private sector. METHODS: Information about dispensed assistive devices and their costs was extracted from (1) the VA's National Prosthetic Patient Database, (2) each site's listing of the VA's Denver Distribution Center cost center in the Cost Distribution Jurisdictional Report, and (3) review of invoices for implanted prosthetics at each study site. We estimated private sector payments by applying Medicare geographically adjusted rates for purchases or rentals, where rates existed, or by inflating VA costs by 30%. RESULTS: The VA spent a total of $30.6 million for prosthetics at the six sites in fiscal year 1999, of which $14.2 million was for items captured in the National Prosthetic Patient Database, $3.4 million for the Denver Distribution Center, and more than $8.1 million for implants. Indirect VA costs were estimated at $4.8 million. Hypothetical private sector payments were estimated at $49.8 million. CONCLUSIONS: Unlike Medicare, VA both contracts to provide assistive devices (through a competitive bidding process) and dispenses devices it has purchased. This approach results in significantly lower expenditures, consistent with other reports. Generalizing these cost savings to other private or federal programs covering assistive devices requires further study. PMID- 12773830 TI - Using the cost distribution report in estimating private sector payments: what adjustments should researchers make? AB - OBJECTIVES: To reapportion Veterans Health Administration (VA) annual expenditures into benefit categories for comparison with estimated payments by private sector providers. METHODS: Total expenditures for six VA medical centers for federal fiscal year 1999 were reapportioned by benefit category using the cost distribution report (CDR). Health benefit categories were based on those of health care insurers. Cost reapportionment was based on CDR data and reviews of source accounting and payroll documents. RESULTS: Actual expenditures for many benefits can be accurately identified and reapportioned using CDR data, but other expenditures were not identifiable in the CDR and required inspection of source documents. Inpatient expenditures amounting to $75,110,094 US dollars and outpatient expenditures amounting to $73,594,284 US dollars were reapportioned into other benefit categories, primarily professional fees. Expenditures for some VA benefits could not be identified because of differences in accounting and clinical practice between the VA and the community. DISCUSSION: Revisions to bring the CDR more in line with private sector payment categories would improve effectiveness for internal VA analyses and external expenditure comparisons. CDR revisions would require changes in recording some clinical workload (eg, rehabilitation and extended care) and classifying residential and domiciliary programs separate from inpatient care. Benefits that were not assigned expenditures for comparison with payments represent a potential liability if the VA were to purchase health care services in the marketplace. Variation among hospitals on expenditures not clearly identified in the CDR was significant and raises questions about the effectiveness of capitated budget methodologies using either the CDR or the decision support system. PMID- 12773831 TI - Applying diagnostic cost groups to examine the disease burden of VA facilities: comparing the six "Evaluating VA Costs" study sites with other VA sites and Medicare. AB - OBJECTIVES: To compare the disease burden of Veterans Health Administration (VA) patients at six study sites with all other VA patients and the Medicare population. DESIGN: A 60% random sample of all VA veteran patients during federal fiscal year 1997 was obtained from administrative databases. A split-sample technique provided a 40% sample (n = 1,046,803) for development and a 20% sample (n = 524,461) for validation. We selected the six study sites from the 40% sample, yielding a total of 50,080 patients in those sites. METHODS: We used Diagnostic Cost Groups to classify patients into clinical groupings based on age, gender, and International Classification of Diseases, Ninth Revision, Clinical Modification diagnoses. The Diagnostic Cost Group model produces relative risk scores that describe patients' expected resource use normalized to the Medicare population. We compared the severity of the six sites with each other and with all other VA facilities and the severity of VA patients with that of Medicare beneficiaries. RESULTS: There were minor statistically significant differences between the study sites and all other VA facilities. Compared with the Medicare population, VA's population was younger and had lower expected resource use (relative risk scores were 1.0 and 0.76, respectively). CONCLUSIONS: Disease burden of the six study sites is representative of all other VA facilities. Although lower relative risk scores suggest that VA patients are healthier than Medicare beneficiaries, when age is taken into account, scores are more comparable. Interpreting the expected resource utilization of the VA population against other benchmarks should be performed carefully. PMID- 12773832 TI - Applying the 3M All Patient Refined Diagnosis Related Groups Grouper to measure inpatient severity in the VA. AB - OBJECTIVES: To assess the severity level of acute inpatient care in the Veterans Health Administration (VA) using the 3M All Patient Refined Diagnosis Related Groups (APR-DRGs) Grouper and compare severity levels in the six study sites with other Veterans Affairs Medical Centers. METHODS: Acute inpatient stays were generated based on bedsection movement information in VA Inpatient Medical SAS data sets from federal fiscal years 1997 and 1998. All nonacute bedsections were excluded. The APR-DRG Grouper generated APR-DRG and severity level for each acute inpatient stay using relevant VA data in a fixed format. Severity and length of stay (LOS) within each major APR-DRG (those accounting for at least 0.5% of all acute inpatient stays or days) were compared between study sites and other centers using z scores. RESULTS: Of 315 APR-DRGs, 63 major groups accounted for more than two thirds of all stays and days of care in both years. The study sites were similar in average patient severity and LOS to other centers for most APR DRGs. For those with significant differences, the six centers had shorter LOS and higher severity. The magnitude of differences was large in LOS and small in severity. CONCLUSIONS: The study sites are generally representative of the overall VA acute inpatient stays. Some adjustments were needed to reflect that the six sites had relatively sicker patients and lower LOS in some of APR-DRGs when resource utilization estimations in the six sites were generalized to the entire VA system. The severity measure of the 3M APR-DRG Grouper can be adapted to the VA controlling for the complicated nature of VA inpatient care. PMID- 12773833 TI - Evaluating VA patient-level expenditures: decision support system estimates and Medicare rates. AB - OBJECTIVES: To make preliminary comparisons of Veterans Health Administration (VA) Decision Support System (DSS) patient-level cost information with Medicare allowable reimbursements. METHODS: For six VA facilities in the Evaluating VA Costs study for federal fiscal year 1999, DSS cost estimates for outpatient inguinal hernia and cataract operations and inpatient stays for chronic obstructive pulmonary disease, simple pneumonia, diabetes, and detoxification were compared with Medicare allowable reimbursement amounts for the same procedures and diagnosis-related groups. Medicare average base payments were adjusted for disproportionate share, capital, and indirect medical education costs. The amounts include Medicare's geographic adjustments for wages and capital. Medicare professional fees were a weighted average of site-specific fees paid for the indicated procedure. RESULTS: For the chosen types of care in fiscal year 1999, average DSS cost estimates were generally higher than estimated Medicare allowable reimbursement amounts, but included different amounts of professional services per discharge or outpatient procedure. The difference was greatest for inguinal hernia repair ($3253 US dollars compared with $1506 US dollars). Two diagnosis-related groups for detoxification (434 and 435) were least comparable between the systems because some VA discharges undoubtedly included both acute and nonacute portions of the hospitalizations, whereas the Medicare rates are for acute stays only. CONCLUSIONS: Researchers and managers need DSS detail records to make any meaningful comparisons of the VA's DSS costs and non-VA reimbursement amounts such as those of Medicare. Non-VA reimbursement estimates should include an average of all professional services, including those of anesthesiologists and consultants. Separating acute and nonacute bedsections in DSS data would improve the VA's capability for comparison. Current information is insufficient for make or buy decisions. PMID- 12773834 TI - Self-management education: more than a nice extra. PMID- 12773835 TI - Having issues with parental psychopathology and children's mental health risk and service use. PMID- 12773836 TI - Can a disease self-management program reduce health care costs? The case of older women with heart disease. AB - BACKGROUND: There is increasing interest in the potential for chronic disease self-management interventions to provide health benefits while reducing health care costs. OBJECTIVES: To assess the impact of a heart disease management program on use of hospital services; to estimate associated hospital cost savings; and to compare potential cost savings with the cost of delivering the program. RESEARCH DESIGN: Randomized, controlled study design. Data were collected from hospital billing records during a 36 month period. Multivariate models were used to compare health care use with cost between treatment and control groups. Estimated differences were then compared with the program costs to determine cost-effectiveness. SUBJECTS: Participants were recruited from 6 hospital sites. Screening criteria included: female, 60 years or older, diagnosed cardiac disease, and seen by a physician approximately every 6 months. The study included 233 women in the intervention group and 219 in the control group. The "Women Take PRIDE" program utilizes a self-regulation process for addressing a problematic area of the heart regimen recommended by each woman's physician. It is tailored to the unique needs of older women. MEASURES: Hospital admissions, in patient days, emergency department visits. RESULTS: Program participants experienced 46% fewer in-patient days (P <0.05) and 49% lower in-patient costs (P <0.10) than women in the control group. No significant differences in emergency department utilization were found. Hospital cost savings exceeded program costs by a ratio of nearly 5-to-1. CONCLUSIONS: A heart disease self-management program can reduce health care utilization and potentially yield monetary benefits to a health plan. PMID- 12773837 TI - Parental depression, child mental health problems, and health care utilization. AB - BACKGROUND: Small controlled studies suggest that depression and other mental health problems are more common in children of parents with depression than in children of parents without depression. OBJECTIVES: This article examines relationships between parental depression and children's mental health problems and health care utilization in a nationally representative household sample of parents and their children. RESEARCH DESIGN: Cross sectional comparisons of sociodemographic characteristics, mental health problems and health expenditures of children whose parents either do or do not report depression in the 1997 Medical Expenditure Panel Survey data. SUBJECTS: A nationally representative sample of children, 3 to 18 years of age (n = 8,360) with one or more parents living in the household. MEASURES: Mental health problems, total health expenditures, and mental health expenditures. RESULTS: Children of parents with depression were approximately twice as likely as children of parents without depression to have a variety of mental health problems and were 2.8 times more likely to use mental health services in adjusted analyses. Among children with health and mental health expenditures, those whose parents report depression had significantly higher mean total annual child health expenditures ($282 vs. $214, t = 3.5, P = 0.0006) and child mental health expenditures ($513 vs. $338, t = 2.0, P = 0.05) than children whose parents did not report depression. CONCLUSIONS: Children of parents with depression are at increased risk for a range of health problems. Parental depression is also related to an increased child health and mental health service utilization and expenditure. PMID- 12773838 TI - Late stage cancers in a Medicaid-insured population. AB - BACKGROUND: Medicaid insurance promotes screening for early stage cancers. However, previous research suggests that Medicaid recipients are at risk for late stage disease. OBJECTIVE: To identify differences in stage of diagnosis between cancer patients enrolled in Medicaid before versus after their disease was identified, as well as differences in diagnostic stage between Medicaid enrollees and other patients. DESIGN: Analyses of a linked database including information from the 1996 and 1997 Michigan Cancer Registry and Medicaid enrollment files. PATIENTS: All persons ages 25 to 64 diagnosed with incident cases of breast, cervical, colorectal, or lung cancer (n = 5852). Patients enrolled in Medicaid before their cancer diagnosis and those enrolled in the same month or after their diagnosis were identified. MAIN OUTCOME MEASURE: Early (in situ, local) versus late (regional, distant, invasive/unknown) cancer stage at diagnosis was modeled using multivariate logistic regression. RESULTS: In each site of disease with the exception of breast, persons who enrolled in Medicaid after a cancer diagnosis were approximately 2 to 3 times more likely to have late stage disease compared with persons who were enrolled in Medicaid before the month of diagnosis. Odds ratios (OR) and 95% confidence intervals (C.I.) were: 1.28 (95% C.I. = 0.95, 1.67) for breast cancer, 2.96 (95% C.I. = 1.85, 4.75) for cervical cancer, 2.08 (95% C.I. = 1.30, 3.33) for colorectal cancer, and 3.40 (95% C.I. = 2.13, 5.43) for lung cancer. Relative to non-Medicaid enrollees, Medicaid enrollees were 2 to 5 times more likely to be diagnosed with late stage disease. CONCLUSIONS: Cancer patients enrolled in Medicaid after their diagnosis were disproportionately likely to have late stage disease relative to patients previously enrolled in Medicaid or non-Medicaid enrollees. PMID- 12773839 TI - The effect of publicly reporting hospital performance on market share and risk adjusted mortality at high-mortality hospitals. AB - BACKGROUND: It is unclear whether publicly reporting hospitals' risk-adjusted mortality affects market share and mortality at outlier hospitals. OBJECTIVES: To examine hospitals' market share and risk-adjusted mortality from 1991 to 1997 at hospitals participating in Cleveland Health Quality Choice (CHQC). RESEARCH DESIGN: Time series. SUBJECTS: Changes in market share were examined for all patients hospitalized with acute myocardial infarction, heart failure, gastrointestinal hemorrhage, obstructive pulmonary disease, pneumonia, or stroke at all 30 nonfederal hospitals in Northeast Ohio. Patients insured by Medicare were used to examine changes in mortality. MEASURES: Trends in market share (proportion of patients with the target conditions discharged from a given hospital) and risk-adjusted 30-day mortality. RESULTS: CHQC identified several hospitals with consistently higher than expected mortality. The five hospitals with the highest mortality tended to lose market share (mean change -0.6%, 95% CI -1.9-0.6), but this was not significant. The only outlier hospital with a large decline in market share had declining volume for 2 years before being declared an outlier. Risk-adjusted mortality declined only slightly at hospitals classified by us as "below average" (-0.8%; 95% CI, 2.9-1.8%) or "worst" (-0.4%; 95% CI -2.3 1.7). However, risk-adjusted mortality at one hospital changed from consistently above expected to consistently below expected shortly after first being declared an outlier. CONCLUSION: Despite CHQC's strengths, identifying hospitals with higher than expected mortality did not adversely affect their market share or, with one exception, lead to improved outcomes. This failure may have resulted from consumer disinterest or difficulty interpreting CHQC reports, unwillingness of businesses to create incentives targeted to hospitals' performance, and hospitals' inability to develop effective quality improvement programs. PMID- 12773840 TI - Improving risk adjustment for Medicare capitated reimbursement using nonlinear models. AB - OBJECTIVES: This article compares a linear risk-adjusted model of medical expenditures for Medicare patients with a model that explicitly account for skewness in distribution of expenditures. METHODS: A model of expenditures and a model of the square root of expenditures, each expressed as linear combinations of risk adjusters, are estimated using data from the 1992 through 1994 Medicare Current Beneficiary Surveys. Five sets of risk adjusters are considered. Each combination of model and set of risk adjusters is tested for linearity, heteroscedasticity, in-sample fit (R2), forecast performance (forecast bias and forecast mean squared error), and overfitting the data. We analyze forecast performance (1)based on forecasts in same year used for estimation, and (2)based on forecasts in the year following that used for estimation. RESULTS: In the first analysis, the model using a square root transformation of expenditures as the dependent variable and the more parsimonious specification of risk adjusters performs best in terms of forecast squared error and overfitting. The untransformed model performs best in terms of forecast bias in each group based on severity of disability, with the exception of the severely disabled for whom the square root model is best. In the second analysis, the square root model performs better than the untransformed model in terms of forecast squared error, but neither model is statistically distinguishable from zero in terms of bias. CONCLUSIONS: Accounting for skewness in expenditures tends to improve precision but not necessarily bias, except among the severely disabled. Adjusting for health status improves risk adjustment. PMID- 12773841 TI - Predicting costs of care using a pharmacy-based measure risk adjustment in a veteran population. AB - BACKGROUND: Although most widely used risk adjustment systems use diagnosis data to classify patients, there is growing interest in risk adjustment based on computerized pharmacy data. The Veterans Health Administration (VHA) is an ideal environment in which to test the efficacy of a pharmacy-based approach. OBJECTIVE: To examine the ability of RxRisk-V to predict concurrent and prospective costs of care in VHA and compare the performance of RxRisk-V to a simple age/gender model, the original RxRisk, and two leading diagnosis-based risk adjustment approaches: Adjusted Clinical Groups and Diagnostic Cost Groups/Hierarchical Condition Categories. METHODS: The study population consisted of 161,202 users of VHA services in Washington, Oregon, Idaho, and Alaska during fiscal years (FY) 1996 to 1998. We examined both concurrent and predictive model fit for two sequential 12-month periods (FY 98 and FY 99) with the patient-year as the unit of analysis, using split-half validation. RESULTS: Our results show that the Diagnostic Cost Group /Hierarchical Condition Categories model performs best (R2 = 0.45) among concurrent cost models, followed by ADG (0.31), RxRisk-V (0.20), and age/sex model (0.01). However, prospective cost models other than age/sex showed comparable R2: Diagnostic Cost Group /Hierarchical Condition Categories R2 = 0.15, followed by ADG (0.12), RxRisk-V (0.12), and age/sex (0.01). CONCLUSIONS: RxRisk-V is a clinically relevant, open source risk adjustment system that is easily tailored to fit specific questions, populations, or needs. Although it does not perform better than diagnosis-based measures available on the market, it may provide a reasonable alternative to proprietary systems where accurate computerized pharmacy data are available. PMID- 12773842 TI - Construction and characteristics of the RxRisk-V: a VA-adapted pharmacy-based case-mix instrument. AB - BACKGROUND: Assessment of disease burden is the key to many aspects of health care management. Patient diagnoses are commonly used for case-mix assessment. However, issues pertaining to diagnostic data availability and reliability make pharmacy-based strategies attractive. Our goal was to provide a reliable and valid pharmacy-based case-mix classification system for chronic diseases found in the Veterans Health Administration (VHA) population. OBJECTIVE: To detail the development and category definitions of a VA-adapted version of the RxRisk (formerly the Chronic Disease Score); to describe category prevalence and reliability; to check category criterion validity against ICD-9 diagnoses; and to assess category-specific regression coefficients in concurrent and prospective cost models. RESEARCH DESIGN: Clinical and pharmacological review followed by cohort analysis of diagnostic, pharmacy, and utilization databases. SUBJECTS: 126,075 veteran users of VHA services in Washington, Oregon, Idaho, and Alaska. METHODS: We used Kappa statistics to evaluate RxRisk category reliability and criterion validity, and multivariate regression to estimate concurrent and prospective cost models. RESULTS: The RxRisk-V classified 70.5% of the VHA Northwest Network 1998 users into an average of 2.61 categories. Of the 45 classes, 33 classes had good-excellent 1-year reliability and 25 classes had good excellent criterion validity against ICD-9 diagnoses. The RxRisk-V accounts for a distinct proportion of the variance in concurrent (R2 = 0.18) and prospective cost (R2 = 0.10) models. CONCLUSIONS: The RxRisk-V provides a reliable and valid method for administrators to describe and understand better chronic disease burden of their treated populations. Tailoring to the VHA permits assessment of disease burden specific to this population. PMID- 12773843 TI - Nonterrorist suicidal deaths involving explosives. AB - Suicidal deaths involving explosives unconnected to terrorism are rare. The investigation of deaths from explosive devices requires a multidisciplinary collaborative effort, as demonstrated in this study. Reported are 2 cases of nonterrorist suicidal explosive-related deaths with massive craniocerebral destruction. The first case involves a 20-year-old man who was discovered in the basement apartment of his father's home seconds after an explosion. At the scene investigators recovered illegal improvised power-technique explosive devices, specifically M-100s, together with the victim's handwritten suicide note. The victim exhibited extensive craniofacial injuries, which medicolegal officials attributed to the decedent's intentionally placing one of these devices in his mouth. The second case involves a 46-year-old man who was found by his wife at his home. In the victim's facial wound, investigators recovered portions of a detonator blasting cap attached to electrical lead wires extending to his right hand. A suicide note was discovered at the scene. The appropriate collection of physical evidence at the scene of the explosion and a detailed examination of the victim's history is as important as documentation of injury patterns and recovery of trace evidence at autopsy. A basic understanding of the variety of explosive devices is also necessary. This investigatory approach greatly enhances the medicolegal death investigator's ability to reconstruct the fatal event as a means of separating accidental and homicidal explosive-related deaths from this uncommon form of suicide. PMID- 12773844 TI - Traumatic subarachnoid hemorrhage and extracranial vertebral artery injury: a case report and review of the literature. AB - The case is presented of a 19-year-old man who was assaulted and died shortly afterward from a large traumatic basal subarachnoid hemorrhage (TBSAH) that arose from rupture of the left vertebral artery, proximal to the point at which the artery penetrated the dura. The literature regarding TBSAH and vertebral artery rupture is reviewed, and a number of points are highlighted: patients with TBSAH may remain conscious for a period of hours after injury, subcutaneous or muscular bruising may be contralateral to the ruptured vessel, fractures of the transverse processes of the cervical vertebrae and significant pathology of the vertebral artery are not typically associated with TBSAH, and rupture of the vertebral artery may be intracranial, junctional, or extracranial. PMID- 12773845 TI - Sudden death during arrest and phencyclidine intoxication. AB - Deaths of individuals being arrested are important and complex medicolegal cases. Conclusions regarding the cause and manner of death for such cases must take into account multiple factors that may have played a role, as well as anticipate the forensic issues that will arise. In this article, we review the deaths of 2 individuals in which phencyclidine intoxication was a factor that contributed to death during arrest. Most cases of sudden death during arrest have involved cocaine intoxication; because phencyclidine's pharmacologic properties are quite different from those of cocaine, these cases allow for comparisons to those factors that may have greater importance. PMID- 12773846 TI - Flow cytometric evaluation of postmortem pulp DNA degradation. AB - During postmortem autolysis, cellular organelles and nuclear DNA break down into their constituent parts. DNA flow cytometric analysis was applied to study the denaturation of splenic cell DNA as a possible method for postmortem interval determination. DNA denaturation continued for 72 hours at a constant rate, with no intact DNA peaks thereafter. The value of using dental pulp tissue for flow cytometric determination of postmortem interval was investigated. The pulps of 57 routinely removed impacted third molars from patients 15 to 30 years of age were obtained. Pulp tissue was removed at 24, 48, 72, 96, 120, and 144 hours postextraction. Debris (degraded DNA) was defined as all signals left of the standardized mean 2n peak and expressed as a percentage of the total number of signals. In contrast to the splenic cell DNA, dental pulp tissue exhibited minimal DNA degradation by 144 hours postextraction, and no constant relation was found between time and DNA degradation during this time. In this study, pulp tissue was found to be unreliable to determine the early postmortem interval but might be of greater value in the later stages. PMID- 12773847 TI - Fatal intravenous injection of potassium in hospitalized patients. AB - We describe 4 cases of fatal intravenous injection of potassium in the hospital setting. These cases illustrate the subtlety of findings in such deaths and remind the forensic pathologist to consider this type of event in sudden, unexpected death of hospitalized patients. PMID- 12773848 TI - Blood-spatter patterns: hands hold clues for the forensic reconstruction of the sequence of events. AB - Biologic and nonbiologic traces on the hands are of particular importance for the forensic reconstruction of shooting incidents; gunpowder residue analysis in particular helps determine whether the gunshot is close range or distant. In crime scene investigation, knowledge about the morphology of bloodstain patterns including gunshot-related back spatter-has increased since various experimental examinations have been performed in the last years; nowadays, these traces are frequently used for forensic crime scene reconstruction. The goal of this study was to deduce the position and orientation of the hands, and therefore the firearm, according to the bloodstain patterns on the hands of the deceased. For this purpose blood-spatter stains on the hands were examined on site in 5 suicides caused by gunshot. In all cases, forensically relevant conclusions regarding forensic reconstruction were enabled through close examination of the spatter marks. Therefore, in shooting incidents, analysis and documentation of blood-spatter findings on the hands is recommended before hands are tested for gunpowder residue or wrapped for the transport of the body. PMID- 12773849 TI - Mortality of children in the Transkei region of South Africa. AB - This study attempted to unfold, perhaps for the first time, the problem of childhood mortality resulting from trauma in the Eastern Cape Province of South Africa. This study was carried out in the Umtata and Ngqeleni magisterial districts, which have a combined population of about 400,000. Most people there have very few resources and have historically relied on money repatriated by migrant workers. In the Transkei region, unemployment is at a very high level: 48.5%. Assault on children is very common in this region, and this may result in death. The aim of this study was to establish the state of deaths resulting from pediatric trauma, and to formulate recommendations that could probably help prevent or reduce these deaths. The objective was to gather epidemiologic information on the victims of pediatric trauma. The study was designed as a descriptive study, using reviews of traumatic deaths in pediatric age groups during the period January 1993 to December 1999. This study was carried out on cases that were brought to the medicolegal laboratory at Umtata General Hospital, Umtata, in the Transkei region of the Eastern Cape Province, South Africa. There were 6181 autopsies conducted from 1993 to 1999. All the medicolegal autopsies were divided into two groups: pediatric (15 years of age or younger) and adult (older than 15 years). Of the autopsies, 89.4% (n = 5587) were in the adult group, and 10.6% (n = 594) were in the pediatric group. Of the pediatric deaths, 64% (n = 383) were related to trauma. The highest numbers were in the 11- to 15 year (n = 146, 38%) and the 6- to 10-year (n = 135, 34%) age groups. Of the children who died of trauma, 112 (28%) were aged 0 to 5 years. Unintentional injuries from motor vehicle accidents were the leading cause of death (59%), whereas intentional injuries (41%) were associated with murder. Nearly a quarter (22%) of pediatric traumatic deaths were due to penetrating injuries: stab (12%) and gunshot (10%) wounds. Most of the pediatric deaths occurred during the festive months of December, January, and April; the death toll during these months was three to four times higher than in the rest of the year. It was concluded that 64% of pediatric deaths in the Transkei region were the result of trauma. This represents 1 pediatric trauma death for every 10 adult trauma deaths. Forty-one percent of the child trauma deaths were due to intentional injuries, usually murder. It is recommended that the government set targets and put into place strategies for the reduction of pediatric deaths due to trauma. The different deaths so called "rule of reversal" could be considered as an indicator of social health. PMID- 12773850 TI - Identification of two fire victims by comparative nuclear DNA typing of skeletal remains and stored umbilical tissues. AB - We describe here our collaborative efforts in identifying 2 fatalities of a fire disaster by using a variety of identification techniques. Postmortem findings in both cases were reinforced using Short Tandem Repeat (STR) DNA technology to establish with a high degree of certainty the identities of 2 child victims. STR markers used in the present study include HUMAMEL, HUMCSFIPO, HUMTHO1, HUMvWA, HUMFES/FPS, HUMF13A01, HUMFOLP23, D8S3O6, HUMFGA, and HUMTPOX. Unambiguous identification was made possible through matching DNA profiles generated from skeletal remains with those from umbilical tissues. These tissues were kept by their mothers in accordance with a Philippine tradition and were submitted for DNA analysis. Of the DNA profiles generated from exhumed bone samples of 21 child victims, comparison with the genetic profiles of children A and B obtained from umbilical tissues showed consistent DNA matches with remains 1756 and 1758, respectively. PMID- 12773851 TI - Laryngeal obstruction by heroin packets. AB - A 28-year-old healthy man collapsed while being arrested by the police for alleged possession of heroin and was found dead on admission to the hospital. Autopsy revealed complete occlusion of the laryngeal opening by a cellophane bag containing 24 packets of heroin powder. PMID- 12773852 TI - Reconstruction of the angle of shot by using computed radiography of the head. AB - In a case of murder by a single .22 caliber bullet fragmented in the head, determination of the trajectory was a key issue in identifying which of 2 windows, 1 of which was associated with 2 suspects, was the site of fire. For this purpose, we processed the computed tomography of the victim's head by using a radiation therapy planning system called Plato, which is routinely used in radiation oncology. This tool provided a 3-dimensional reconstruction of the cranio-cerebral lesions and a spatial location of bullet and bone fragments. In this article is described the more intuitive relationship with forensic and ballistic data obtained from the 3-dimensional visualization and the physical and mathematical analysis used to calculate the angle of shot. PMID- 12773853 TI - Four major disasters in Aquitaine, France: use of odontologic techniques for identification. AB - The 4 mass disasters that have affected the Aquitaine region (France) in the previous 15 years are related through the specificity and the efficiency of dental methodology in identifying the victims. The results point to the decisive influence and the efficiency of dental methodology for identification of bodies that are burned or disfigured after an accident or a prolonged period in water. PMID- 12773854 TI - A case of electrocution during illegal fishing activities. AB - The passage of electric current through the human body causes variable harm, ranging from loss of consciousness to death caused by paralysis of the bulbar nerve centers. This report describes a fatal case caused by an illegal fishing practice involving stunning fish with electric shocks produced by an electric generator carried through the water inside the carcass of an old refrigerator to keep it afloat. This occurrence is unusual because of the circumstances giving rise to the use of electric current and the cause of death, which resulted from the combination of the electric current produced by the generator and the saltwater, bringing about death by drowning. PMID- 12773855 TI - Determination of sex and race from the subpubic angle in Ugandan subjects. AB - The accurate determination of sex and race are important tools to forensic and physical anthropologists. In this study, the sex and race of 205 adult Ugandans were determined from the antero-posterior radiographs of the pelvis by measuring their subpubic angles. The angle ranged from 50 to 140 degrees with a mean of 93.86 degrees and standard deviation (SD) of 21.12 degrees for males and 75 to 155 degrees with a mean of 116.11 degrees (SD, 17.79 degrees) for females. The angle was significantly wider in women than men (P < 0.05), as indeed has previously been shown in other population groups studied. Using the demarking point method, 31.82% of Ugandan men and 10.53% of Ugandan women could be accurately sexed. However, more Malawians of both sexes previously studied had lower values for the angle and could be more accurately sexed. This study has also documented regional and racial variability of this angle among different population groups previously documented. Using the watershed subpubic angle derived from the means of this study and those of previously documented studies, 63% of Ugandans and 71% of Malawians could be accurately assigned to the black race. This method is recommended to physical and forensic anthropologists in the developing world. PMID- 12773856 TI - Death during theft from electric utilities. AB - Utilities that supply power are subject to theft of service and theft of hardware used to provide service. Individuals who try to steal from a power utility risk electrocution. We conducted a retrospective study of all individuals examined by the Jefferson County Coroner Medical Examiner Office from January 1981 through December 2001 and found that 8 individuals died trying to steal from a power utility. All decedents were men, with an average age of 33 years. Seven decedents were stealing copper, and 1 was stealing electricity. Only 1 decedent survived long enough to be admitted to a hospital. Five decedents were intoxicated, 3 with ethanol, 1 with cocaine, and 1 with both cocaine and ethanol. Seven decedents sustained electrical burns in keeping with the high voltage to which they were subjected. The unburned decedent died of laceration of the aorta at its origin from the heart, an injury sustained when the decedent fell from the power pole. All 8 decedents attempted their theft during daylight. Items found in the decedent's pockets, the presence of a ladder nearby, or the presence of electrical tools around the body may help to establish the attempted act of theft. PMID- 12773857 TI - Choking death on a live fish (Dicologoglossa cuneata). AB - A choking death of a healthy fisherman, who put a type of live sole between his jaws to free up his hands so that he could collect more fish to put into his basket, is described. The fish squirmed into the larynx and upper trachea and the attempts to rescue the man by his colleagues who used pliers did not succeed, and the man died. Other published cases are reviewed, and risk factors and rescue possibilities discussed. PMID- 12773858 TI - Analysis of neck injuries in hanging. AB - Retrospective study of suicidal hangings was made on 175 cases (133 male victims, 42 female victims) for a 5-year period. The mean age was 47.33 +/- 17.51 years. The sample was divided in 4 groups according to the position of the ligature knot (anterior, right, left, and posterior hanging). The authors analyzed all visible injuries of soft tissues and bones and cartilage of the neck, and in 150 cases (85.7%), we established that there was at least one injury of these structures (hematoma or fracture for example). The most frequent injury was to sternocleidomastoid muscles. Fracture of throat skeleton was detected in 119 cases (68%). The proportion of fractures increases with age of the deceased. There was no clear correlation between frequency of neck injuries and type of hanging. PMID- 12773859 TI - Forensic medicine in Bordeaux in the 16th century. AB - In the 15th and 16th centuries, the Renaissance was accompanied by a real cultural revolution in Europe and France. Montaigne, the Bordeaux humanist and writer, was highly involved in this movement, particularly by his thinking on medicine, physicians, and illness. The 2 forensic reports presented are the oldest known testimonies of forensic medical activity in Bordeaux in the 16th century. They concern a visit to prison in 1556 and the solemn transfer of the body of a man hanged in 1579. The authors also describe how necropsies were performed before 1573 in Bordeaux, the year in which medical studies were reformed. However, the first official teaching of forensic medicine in Bordeaux (Ecole Royale de Medecine) began only in 1814. PMID- 12773860 TI - Evaluation of 15 biparental STR loci in human identification and genetic study of the Kannada-speaking groups of India. AB - Thirteen tetranucleotide and 2 pentanucleotide repeat units were analyzed in 167 unrelated Kannada-speaking individuals belonging to 3 important communities, namely, Kuruva, Bhovi, and Christians, residing in different districts of Karnataka, India. Allele frequency data obtained from the analysis of 15 short tandem report (STR) markers of the subpopulation groups included in the study were observed to be similar, indicating a common ancestry or gene flow among these communities. Departures from Hardy-Weinberg equilibrium were observed in Kuruva population at locus D5S818 and D18S51 and at locus CSF1PO in the Christian community. The data of these communities were analyzed with allele frequency data of 4 other populations from Karnataka, India-Iyengar Brahmin, Gowda, Lingayat, and Muslim-to compute the combined power of discrimination, ranged from 0.962 to 0.974, with negligible difference between populations. The combined power of exclusion, however, remained constant at 0.999 for all populations evaluated in the study. Thus, the 15 markers selected for this study were found to be highly suitable in human identification and for providing information on genetic polymorphism. PMID- 12773861 TI - Pulmonary embolization by chorionic villi causing maternal death after a car crash. AB - Throughout the last century, there has been a marked decline in obstetric maternal deaths, resulting in an increase in the proportion of nonobstetric deaths among pregnant women. Trauma, in particular, has become a leading cause of maternal death. We report the case of a 20-year-old primigravid woman who was involved in a motor vehicle crash at 36 weeks gestation. The woman developed abruptio placentae, followed by disseminated intravascular coagulation, adult respiratory distress syndrome, and shock, and died the day after the crash. Widespread pulmonary embolization by chorionic villi was identified at autopsy. This report discusses traumatic maternal deaths, with emphasis on the differences in injury pattern observed in pregnant trauma victims in comparison with other adults. It is important that the pathologist be aware of these problems so that an accurate cause of death can be identified in cases of maternal death after trauma. Also discussed is the relationship between trauma and placental abruption and the mechanism of death in the patient. To the authors' knowledge, this is the first reported case of extensive embolism of chorionic villi to the lungs after trauma. PMID- 12773862 TI - Case report of a homicide by a shotgun loaded with unusual ammunition. AB - Reported is the case of a man killed by a neighbor with a shotgun loaded with a unusual ammunition. The cartridge contained several shots chained together by metallic wires. Such ammunition has been used for hunting purposes but is now banned, so it can be handmade only. The joined pellets produce a wound with features that are different from those of a wound made by a regular load of pellets. Several interpretive problems are caused by the unknown number of pellets contained in the shell and the effects of the wire holding the shot together. PMID- 12773863 TI - Custody-related deaths in Durban, South Africa 1998-2000. AB - Custody-related deaths have become a major human rights issue in South Africa since the 1994 democratic elections and led to the formation of the Independent Complaints Directorate who, in April 2000, had recorded 2174 custody-related deaths since their inception. A 3-year retrospective descriptive study on all custody-related death cases at a Durban Medico-legal mortuary from January 1998 to December 2000 was undertaken. The study included deaths that occurred during arrest and deaths within the South African Police Services holding cells. Deaths of prisoners within the Correctional Services and psychiatric institutions were excluded. A total of 117 cases were reviewed, of which 88 (75.21%) were due to police shootings, 11 (9.40%) were due to assault by the police during arrest, and 3 (2.57%) were due to assault by police dogs. There were 102 (87.18%) "police action" related deaths and 16 (13.68%) police custody deaths, of which 7 (5.99%) were due to suicide by hanging and 2 (1.71%) due to natural causes. The results from this study differ significantly from published reports from other countries, with respect to the high numbers of deaths caused by police action. Prevention of custody-related deaths should be recognized as an area of concern and should be given greater priority by the government and the South African Police Services. PMID- 12773864 TI - Medicolegal diagnostic value and clinical significance of traumatic incomplete tears of the basilar artery. AB - Ruptures of arteries of the vertebrobasilary system are relatively frequent in medicolegal practice, and their origin may be both natural and violent. Tears that affects the whole thickness of the basilar artery cause subarachnoid hemorrhage (SAH), with an often rapid fatal outcome. 1-3 However, in some situations, arterial tears may be incomplete, involving the intima or both the intima and the media, but with preserved adventitia. 1, 4 Although such incomplete tears are not the source of immediate subarachnoid bleeding, their presence may be important from both a medicolegal and a clinical point of view. The aim of this article is to point out the significance of incomplete tears of basilar artery as a possible diagnostic sign of traumatic origin of SAH as well as a certain mechanism of injury, which involves forcible hyperextension and rotational movements of the head. The authors also describe their method of performing longitudinal section of the basilar artery, both at autopsy and for histologic examination, which is convenient for identifying multiple transversal incomplete tears of this blood vessel. The article is based on the analysis of three cases from the autopsy material of the Institute of Forensic Medicine in Belgrade. PMID- 12773865 TI - Suicide in children and adolescents at a province in Turkey. AB - Despite the well-documented increases in the rate of completed suicide among children, accurate knowledge of the characteristics of these suicides is limited. We examined all suicides by children and adolescents in Diyarbakir during 1999 and 2001. All of the cases were analyzed regarding the age, sex, method and location of suicide, time of year, and predictive factors. There were 56 children aged 11 through 19 years. Adolescents between 15 and 19 years of age constituted the overwhelming number (87%) of all the childhood suicides. Female children and adolescents predominated among the suicide victims (71%). The most frequent means of suicide were firearms (43%), hanging (28%), and jumping (16%), respectively. The majority of suicides occurred at the decedent's home (88%). Reasons identified were mainly psychiatric disorders, followed by troubles within the family. A prior suicide attempt or a note that was left by the decedent and elucidated the reason for the suicide was rare. There was no seasonal difference in rates of suicides. PMID- 12773866 TI - [Cerebral ischemic events and anti-cancer therapy]. AB - Antineoplastic agents have been associated with cerebral hemorrhage, infarction and cerebral venous thrombosis. Infarctions have been reported in association with L-asparaginase, cisplatinium, methotrexate and 5-fluro-uracil. The mechanisms by which antineoplastic agents may lead to stroke include endothelium toxicity and abnormalities of coagulation factors. PMID- 12773867 TI - [At the crossroads between developmental and degenerative diseases: the cerebellar disorders of early infancy. Classification and practical approach]. AB - The developmental characteristics of the cerebellum, including its histogenesis which persists well beyond birth, explain, at least in part,why the mechanisms of cerebral disorders of infancy remain equivocal. The nosology of certain congenital ataxias, especially those with cerebellar hypoplasia, remains ambiguous, at the crossroads between early degenerative disease and congenital non-progressive anomalies. We have revisited the clinical approach to the most frequent situations: (1) the careful dysmorphology work-up must search for any element of various recognizable syndromes, especially those transmitted by autosomal recessive inheritance. An update of list of such syndromes is provided. (2) Cerebellum imaging must be obtained as early as possible and re-documented with a long-term follow-up. Emerging 3D techniques should help improve morphological evaluation. (3) One the contrary, a complex biochemical work-up, looking for metabolic diseases, is required only when the clinical and radiological evaluations provide unusual data. (4) Mental status is always the most relevant element of prognosis. t is frequently compromised, including in congenital non-progressive ataxia with normal imaging. Beyond the classical strategies, the genetic approach must take into consideration possible phenotypic homologies with natural or experimental animal models. This approach is illustrated by the recent discovery of mutations with the human homolog of the Reeler gene in a subset of cerebellar agenesis associated with other dysgenetic elements. PMID- 12773868 TI - [Relapsing-remitting inflammatory disease of the central nervous system with normal MRI: multiple sclerosis or phenocopy in a series of 15 patients]. AB - Multiple sclerosis is a demyelinating disease of central nervous system. Although many sub-types and clinical forms are identified, diagnosis is clearly related to the detection of MS lesions on brain MRI. We report data of 15 patients admitted in Nice for suspicion of MS after clinical relapsing-remitting or progressive symptoms. Extensive screening tests (i.e blood sample, CSF, MRI, spectroscopy) were performed at onset and at each relapse. All patients had normal-appearing white matter on spinal cord and brain MRI. Nevertheless, 11 patients can be considered as MS according to McDonald criteria. PMID- 12773869 TI - [Prevalence of dementia and Alzheimer's disease among subjects aged 75 years or over: updated results of the PAQUID cohort]. AB - The progression of the prevalence of dementia in developed countries will increase the difficulties of medical and psycho-social management of demented patients and their family. The aim of this study was to estimate the prevalence of dementia among subjects aged 75 years and over. These subjects have been followed-up ten years after the baseline examination of the PAQUID study, a prospective population-based cohort study on normal and pathological aging after 65 years. A total of 1,461 subjects, aged 75 years and over, have been visited. The prevalence of dementia was estimated to be at 17.8 p.cent. About 38.5 p.cent of these demented subjects ware living in an institution, which leads to the fact that more than the two thirds of the subjects living in an institution were demented persons. Alzheimer's disease was the main etiology of dementia (79.6 p.cent). Based on the French population census, currently there would be about 769,000 demented people aged 75 years or over in France. Dementia accounts for 72 p.cent of the potential need for the French subvention for dependent persons, called Allocation Personnalisee d'Autonomie P. These results point out the importance of regarding dementia as a Public Health priority and of developing necessary measures for caring for these subjects, whether residing in their home or an institution. PMID- 12773870 TI - ["Progressive acalculia": a variety of focal degenerative atrophy affecting number processing]. AB - A 72-year-old man experienced increasing difficulties in calculating and processing numbers. He made many syntactic errors when he read and wrote numbers, both in arabic and number-word forms. Transcoding between arabic numerals and number-words was severely impaired. His calculation abilities were grossly impaired. He could not perform even simple multiplications, but additions and subtractions were preserved. He was unable to recite multiplication tables or the alphabet. In contrast, numerosity judgment, magnitude comparisons,and semantic evaluation of numbers were intact. The patient's deficit could be summarized as an Impairment in all tasks involving numbers in a verbal format, corresponding to the so-called "verbal anarithmetia'." It was noteworthy that verification of operations, including multiplications, was accurately performed. For example, the patient could not solve a multiplication like 8 x 9, but could estimate that 9 x 2=1 1 was closer to the exact result than 9 X 2=17, or could state that 8 x 2=16 was exact even though, at the same time, he expressed verbally the terms of the corresponding addition "eight plus two is ten." Other cognitive deficits were detected by neuropsychological tests which demonstrated mainly anomic aphasia, dysorthographic agraphia without alexia, impairment of short term and episodic memory, digital agnosia. However, the patient could not be considered to have demon-eti due to preserved self-sufficiency, except for tasks requiring calculation abilities. During the two years of follow-up, the impairment in processing numbers and calculation remained predominant over other deficits. Cerebral MRI showed an atrophy of the left temporal and parietal lobes, and SPECT study showed a reduction of regional cerebral blood flow of the left hemisphere. This original clinical presentation of a cerebral degenerative disease could be described as "primary progressive acalculia" whatever the underlying primary pathological process. PMID- 12773871 TI - [Early diagnosis of bacterial brain abscesses: interest of diffusion-weighted MRI]. AB - Three cases of bacterial brain abscesses, in immunocompetent patients, are reported. In all these cases, the diffusion-weighted magnetic resonance (MRI) with apparent diffusion coefficient (ADC) map has permitted an early diagnosis and a rapid treatment. This emergency MRI showed in the three cases a low signal on TI-weighted images, a high signal on T2-weighted and echo-planar images, and a decrease of ADC (0.36- 0.49 x 10(-3) mm2/s). So, this new MRI technique provides an available and rapid element in the brain abscess diagnosis which often remains a complex clinical and radiological diagnosis. PMID- 12773872 TI - [Muscle granuloma: anatomoclinical correlation and immunohistochemistry in seven cases]. AB - Granulomatous inflammation is infrequently observed in muscle biopsy. We report a series of 7 patents presenting with granulomas in muscle. Two of them had a history of sarcoidosis In 4 other cases, muscle Involvement revealed systemic sarcoidosis. Among the 6 cases of sarcoidosis, we observed 2 with acute myositis and 4 chronic forms. The last patient presented with polymyositis in association with melanoma. In sarcoidosis, muscle biopsy showed a granulomatous inflammation of varying intensity, which was generally associated with mononuclear inflammatory cells. Most of granulomas were located in the perimysium and the endomysium and necrosis was absent. Inflammatory cells were predominantly macrophages and CD4 positive lymphocytes. On the contrary, in the case of paraneoplastic polymyositis,granulomas were rare, most of inflammatory cells were CD8 positive lymphocytes and numerous areas of necrosis were observed. Class I MHC molecules were expressed on the membrane of muscle fibers. As a general rule, requisite examinations must be performed to search for sarcoidosis in patients exhibiting granulomas on muscle biopsy. PMID- 12773873 TI - [Melanocytic meningitis and large congenital melanocytic naevus: neurocutaneous melanosis]. AB - Neurological symptoms in a patient with large congenital melanocytic naevus are highly suggestive of cerebromeningeal melanoma metastasis. The presence of melanocytic cells in cerebrospinal fluid confirms this diagnosis If their malignant nature is shared with cutaneous naevocytic cells. Conversely, neurocutaneous melanosis is diagnosed when benign melanocytosis meningitis is found in patients with multiple and/or large congenital melanocytic naevus, whether cutaneous naevus cells are benign or not, or when cerebrospinal fluid cells are malignant with benign cutaneous melanocytic naevus. We report the case of a young man aged 19 presenting with multiple and large congenital melanocytic naevus who experienced transcient neurological signs and increased intracranial pressure. Cerebral neuroimaging evoked meningeal infiltration which benign melanocytic nature was supposed on CSF analysis and confirmed by necropsy findings, only 3 month after neurological onset, leading to neurocutaneous melanosis diagnosis. This rare neuroectodermal dysembryoplasia finds expression in various neurological signs, depending on patient's age and leptomeningeal and/or cerebral proliferation localization. Lumbar puncture, cerebral scanography and MRI may help diagnosis, but only histological examination can prove neurocutaneous melanosis, more often by necropsy because of poor prognosis. PMID- 12773874 TI - [Pathological gambling behavior in a patient with Parkinson's disease treated with levodopa and bromocriptine]. AB - The occurrence of a pathological gambling behavior in a 61-year-old patient with idiopathic Parkinson's disease treated with dopaminergic drugs is reported. This is the first case reported with bromocriptine. The main characteristics and the mechanism of this recently described and a unexpected, adverse drug reaction are discussed. PMID- 12773875 TI - [Calcified brain metastases of a pulmonary neuroendocine carcinoma]. AB - We describe an unusual case of pulmonary neuroendocrine carcinoma of intermediate cell type, revealed by calcified metastatic nodules of the brain in a 49-year-old man. Histological examination of the brain lesions disclosed necrosis and calcification in the center with small and medium-sized carcinoma cells. Pathophysiological mechanisms of such calcifications in metastasis lesions are reviewed. PMID- 12773876 TI - [Cerebral vasculitis secondary to Toxocara canis and Fasciola hepatica co infestation]. AB - Neurological complications of cosmopolitan parasitosis are rare. We report a case of a cerebral and systemic vasculitis secondary to a visceral larva migrans syndrome associated with a Toxocara canis and Fasciola hepatica co-infestation. PMID- 12773877 TI - [Thrombus of the left lateral sinus spreading to the internal jugular vein]. AB - Cerebral venous thrombosis is an uncommon event which presents a wide spectrum of sometimes extraneurological signs different from the classical clinical presentation. We report the cases of two middle-aged women who developed thrombosis of the left lateral sinus spread-ing to the internal jugular vein from the sigmoid sinus. The time course of the symptoms suggested that intracranial thrombosis occurred first. No infectious or neoplastic local disease could be found but both women were taking oral contraceptives. Medical treatment led to good reperfusion of the intracranial sinuses but occlusion of the jugular vein persisted despite prolonged oral anticoagulants. Long-term outcome was favorable with residual benign epilepsy in one patient, and occurrence of an arteriovenous fistula in the other. PMID- 12773878 TI - [Isolated neurosarcoidosis without systemic signs]. AB - Sarcoidosis is a multisystemic disease which involves the nervous system in 5 to 15 p.cent. Neurosarcoidosis without signs of systemic disease is rare and may be difficult to diagnose. We report a case of a 61 year-old patient with a pseudotumoral neurosarcoidosis and along evolution of 25 years without systemic signs. In such cases, histological analysis is rewarding. PMID- 12773879 TI - [Tick meningoencephalitis]. PMID- 12773880 TI - [Antineuron antibodies and paraneoplastic neurological syndromes]. PMID- 12773882 TI - [Proceedings of the 8th Meeting of Therapeutics and Neurology: Parkinson Disease. 7-8 December 2001, Paris, France]. PMID- 12773881 TI - [Dysarthria?]. PMID- 12773883 TI - [Recent neuropathology of parkinsonian syndromes]. AB - The understanding of the molecular mechanisms underlying Parkinson's disease, progressive supranuclear palsy, and multiple system atrophy has made significant progress in the recent years. Lewy body appears to be principally made of alpha synuclein, a presynaptic protein. It also contains ubiquitin and some components of the proteasome: this suggests that alteration of protein catabolism may be involved in its formation. In favor of this hypothesis, it should be noted that Parkin, a protein that is mutated in autosomal recessive Parkinson disease, is a ubiquitin ligase. Immunohistochemistry has shown that alpha-synuclein accumulates not only in the cell body of the neurones (Lewy body) but also in their processes (Lewy neurites); it has emphasized the severity of the pathology in the nucleus basalis of Meynert, amygdala, CA2-3 sector of the hippocampus and cerebral cortex. Cortical Lewy bodies are not considered any more the marker of dementia with Lewy bodies: they are, indeed, found in true Parkinson disease cases. In progressive supranuclear palsy, 4 repeats tau accumulates in the cytoplasm of neurones and glia. At electron microscopy, the accumulation is made of straight filaments. It involves not only the neurones (where it is the main constituent of the neurofibrillary tangles) but also the glia. Astrocytic tuft is to day considered the morphological marker of progressive supranuclear palsy. Tau protein accumulates in the cell body of the oligodendrocyte as a "coiled body"; the protein is also integrated in the myelin sheath, when the cytoplasm of the oligodendrocyte wraps around the axon. This explains the numerous "threads" that are visible in cases of progressive supranuclear palsy. Striato-nigral degeneration, sporadic olivo-ponto-cerebellar atrophy and primitive orthostatic hypotension are various clinico-pathologic aspects of the same disorder: multiple system atrophy. It is also characterized by a morphological marker: the accumulation of alpha-synuclein in the cytoplasm of glial cells, particularly oligodendrocytes. The term synucleinopathy has been proposed to describe both idiopathic Parkinson disease and multiple system atrophy. The reason explaining the cellular topography of alpha-synuclein accumulation, neuronal in Parkinson disease, glial in multiple system atrophy is still unknown. PMID- 12773884 TI - [Diagnostic criteria of parkinsonian syndromes]. AB - Parkinson's disease constitutes 75p. 100 of all causes of parkinsonism. The disease is defined clinically by the UK Parkinson's Disease Brain Bank (UKPDSBB) which exclude secondary causes of parkinsonism and atypical primary parkinsonian syndromes. Any atypical sign, also commonly known as "red flags", should make the clinician aware of an alternative diagnosis: multiple system atrophy, progressive supranuclear palsy, corticobasal degeneration and parkinsonism associated with dementia such as dementia with diffuse Lewy body disease. Despite progress in designing validated clinical diagnosis criteria, such diseases remain difficult to diagnose in their early course. PMID- 12773885 TI - [Progressive supranuclear palsy]. AB - Progressive supranuclear palsy (PSP) is a parkinsonian syndrome 20 to 30 times less common than Parkinson's disease. PSP and Parkinson's disease share certain symptoms, but also present distinctive features. The histopathological features of PSP are highly specific and enable certain diagnosis. Since no biological marker has been identified, clinical signs must be recognized to establish the probability of the histopathological diagnosis. The NINDS program distinguishes two categories: probable PSP and possible PSP with distinctive characteristics. The sensitivity of probable PSP is only 50 p. 100, but the specificity and positive predictive value are 100 p. cent. For possible PSP, specificity is 83 p. cent (93 p. cent if diagnostic errors are included) and the positive predictive value is lower (83 p. cent). Although there number is smaller, patients with probable PSP are retained for research on new treatments. There have been few, non-controlled therapeutic trials in PSP. No drug and no surgical procedure has been demonstrated to be highly effective. Neuroprotection is a new avenue of research which requires knowledge of the natural course of clinical incapacity. A longitudinal study of 50 patients with probable PSP has provided mid-term results on symptom onset and on the development of three key signs, detected with the standard Unified Parkinson's Disease Rating Scale: incapacity to stand alone without help, incapacity to speak intelligently, incapacity to eat (tube feeding). The Kaplan-Meier curves show that the mean duration of the disease before development of one of these signs is 50 months. Research on protocols designed to stop disease progression should be centered on this period. PMID- 12773886 TI - Multiple system atrophy. AB - Multiple system atrophy (MSA) is a sporadic neurodegenerative disorder that usually manifests in the early fifties and progresses relentlessly with a mean survival of 9 years. Clinically, MSA is dominated by autonomic/urogenital failure which may be associated with either levodopa unresponsive parkinsonism in 80p.cent of cases (MSA-P subtype) or with cerebellar ataxia in 20p.cent of cases (MSA-C subtype). Pathologically, MSA is characterized by a neuronal multisystem degeneration and abnormal glial cytoplasmic inclusions containing alpha-synuclein aggregates. Pharmacological treatment of motor features is disappointing except for a transient L-Dopa response in a minority of MSA-P patients. In contrast, autonomic and urogenital features of MSA should be identified early on because they can be treated effectively in many instances. To improve the outcome of this devastating disorder which causes early severe and progressive disability more research into etiology, pathogenesis, early diagnosis and therapeutic management is necessary. Several research groups have been formed in Europe (EMSA-SG, NNIPPS) and USA (NAMSA-SG) to address these unmet needs. PMID- 12773887 TI - [Secondary parkinsonian syndromes]. AB - Secondary parkinsonism (SP) is caused by an identified structural, toxic, or metabolic mechanism. The first group of SP syndromes includes disorders caused by a primary pathological process in the brain. The second group includes disorders with a primary extrinsic or ubiquitous causal factor resulting either from reversible dysfunction of the basal ganglia or structural disorders related to the predominant or selective vulnerability to aggression of the basal ganglia. The pathophysiological aspects SP are developed here, particularly vascular syndromes and syndromes secondary to hydrocephalus. In both conditions, it is rare that SP resembles Parkinson's disease. The pathophysiology is poorly understood, but would involve, in most cases, corticostratial and/or thalamocortical connections in the basal ganglia system. For the clinician, the practical problem is frequently to determine whether a patient's parkinsonism can be related to an identifiable cause or whether there is an associated Parkinson's disease or other degenerative disease. When both a known cause of parkinsonism and Parkinson's disease are present, the cause may be asymptomatic, reveal Parkinson's disease, have an aggravating effect on the underlying disease, or modify its clinical expression. PMID- 12773888 TI - [Neuropsychological evaluation can contribute to the diagnosis of movement disorders]. AB - Neurodegenerative diseases of the extrapyramidal system can be difficult to identify, particularly those with discrete symptoms or a non-specific or polymorphous complex motor expression. Nevertheless, most of the patients with these diseases present a particular cognitive pattern so the neuropsychological evaluation can contribute significantly to diagnosis. Progressive supranuclear palsy, corticobasal degeneration, Huntington's disease, and Lewy body dementia can be easily identified on the basis of their distinctive motor and cognitive features. Clinical manifestations may be less specific in other diseases such as striatonigric degeneration or idiopathic Parkinson's disease. We present here the characteristic neuropsychological features of each of these diseases and their contribution to diagnosis. PMID- 12773889 TI - [Is functional imaging useful in the clinical evaluation of Parkinson's disease?]. AB - Functional imaging is mainly used for clinical research. However, if available, this technique could help the clinical diagnosis. First, because it is possible to confirm the loss of nigrostriatal Dopaminergic neurons using [(18)F]-L-Dopa with PET or [(123)I]-FP-CIT with SPECT. Second, distinguishing idiopathic from atypical parkinsonism might be possible using the assessment of striatal Dopaminergic D2 receptors or the measurement of brain metabolism. However, there has been no prospective study to confirm this hypothesis and functional imaging is only performed to exclude patients with atypical parkinsonism from the waiting list for neurosurgical treatments. PMID- 12773890 TI - [Neurophysiology of parkinsonian syndromes]. AB - Neurophysiological investigations may contribute to establish the diagnosis of different parkinsonian syndromes such as multiple system atrophy, progressive supranuclear palsy and corticobasal degeneration, which are often suspected in case of atypical parkinsonian syndrome. They provide a means of objective analysis of certain symptoms (myoclonus for example) and identify the pathophysiological processes underlying these parkinsonian syndromes. This article reviews especially the following electrophysiological tests: autonomic nervous system analysis (sympathetic sudomotor skin response, R-R interval variation, electromyography of the external anal sphincter), surface electromyography and movement recording to investigate myoclonus, somatosensory and motor evoked potentials, brainstem reflexes (mainly the blink reflex), startle reaction, monosynaptic and long latency reflexes and finally the premotor potentials (contingent negative variation, Bereitschaftspotential) and movement related desynchronization). PMID- 12773891 TI - [Contribution of oculomotor examination for the etiological diagnosis of parkinsonian syndromes]. AB - Exploration of ocular motricity can be helpful for diagnosis in certain parkinsonian syndromes. Oculomotricity is perturbed in Parkinson's disease (PD) and in multiple system atrophy (MSA). The minimal anomalies, sometimes observed in these conditions, both clinically and on oculomotor recordings, cannot contribute significantly to diagnosis. In corticobasal degeneration (CBD), infraclinical oculomotor anomalies can be identified on oculomotor recordings and are relatively specific to atypical parkinsonian syndrome: particularly long latency of ocular saccades related to posterior parietal involvement. The most significant contribution is observed for progressive supanuclear palsy (PSP) where the oculomotor anomalies are evident at clinical examination and are even the cardinal signs of the condition: particularly paralysis of vertical ocular saccades (downbeat and upbeat) or downbeat saccades, ocular pursuit also being perturbed but not reflex movements (preservation of oculocephalic reflexes). This dissociation between palsy of saccade and pursuit movements and preservation of oculocephalic reflexes is the sign of the supranuclear origin of the oculomotor disorder in PSP. In early stage PSP before clinical expression, requiring oculomotor recordings to be recognized, it is relatively specific in the context of degenerative parkinsonian degeneration. PSP also involves other less specific anomalies such as a perturbation of the antisaccade movements (related to a frontal syndrome), anomalous ocular pursuit (becoming saccadic) and presence of square wave. A rigorous clinical examination of oculomotor function at a more advanced stage of PSP, or oculomotor recordings at an early stage of PSP or CBD, can thus contribute significantly to the diagnosis of both conditions. PMID- 12773892 TI - [Dopaminergic treatment and parkinsonian syndromes]. AB - Chronic dopamine treatment usually provides partial and temporary improvement of extrapyramidal signs in about 40p.cent of the patients with multiple system atrophy. Exceptionally, dopamine agonists may provide a significant and persistent improvement in progressive supranuclear palsy. For patients with Lewy body dementia, levodopa often provides a significant improvement of the extrapyramidal syndrome in about 70p.cent of the treated patients. Dopamine treatment generally has no effect on secondary extrapyramidal syndromes caused by vascular parkinsonism or neuroleptics. Antiparkinsonian treatment can cause several complications, particularly degenerative parkinsonian syndromes with an increased frequency of cognitive disorders and dysautonomia. Consequently, antiparkinsonian agents should only be proposed within the framework of a differential diagnosis between parkinsonism and idiopathic Parkinson's disease then continued only if the risk/benefit ratio, systematically evaluated for each patient, is favorable. PMID- 12773893 TI - [Treatment of non-dopamine-dependent signs in parkinsonian syndromes: evaluation and results]. AB - With dopaminergic systems, non dopaminergic neurotransmission probably plays a major role in parkinsonian syndromes (Multiple System Atrophy, Progressive Supranuclear Palsy, Pure Autonomic Failure, Cortical basal degeneration, Lewy Body Disease). A better understanding of the pathophysiology of these syndromes led to the development of molecules that interact with non dopaminergic systems. Thus, freezing, gait and balance disorders, dysautonomia and neuropsychiatric disorders are likely to benefit from specific treatments. However, due to methodological difficulties related to the evaluation of such molecules, controlled trials are rather rare and the results are often partial and sometimes unclear. PMID- 12773894 TI - [Neuroprotection and neurodegenerative parkinsonian syndromes]. AB - The diffuse nature of the lesions in neurodegenerative parkinsonian syndromes explains the inefficacy of symptomatic treatments and the potential interest of neuroprotector treatments that could slow down or even prevent neuron degeneration in structures involved in the degenerative processes. As these syndromes share preferential degeneration of the substantia nigra with Parkinson's disease it is logical to hypothesize that the same mechanisms of neuron death are involved. The responsibility of an exotoxin, with a mechanism of action that would be similar to that of MPTP and/or rotenone, appears to be implicated only in progressive supranuclear palsy (PSP): this is suggested by the "guadeloupean parkinsonean" syndrome. There is no evidence demonstrating an exotoxin in corticobasal degeneration (CBD), which might play an anecdotal role in rare cases of multiple system atrophy (MSA). There are rare cases of PSP, sometimes with autopsy proof, generally with autosomal dominant inheritance, but in the much larger number of sporadic cases there is an undeniable genetic susceptibility linked with certain polymorphisms of the tau protein gene. Genetic susceptibility plays a much less pronounced role in CBD. There is no argument however in favor of a genetic factor in MSA. A few arguments suggest that oxidative stress is involved in PSP and MSA, or even CBD, but no evidence of a primary effect. Perturbed mitochondrial metabolism is possible in PSP. Undeniable proof of the effect of inflammation, excitotoxicity, and apoptosis remains to be presented. We now have several compounds which could affect different phases of neurodegeneration. Identifying the precise cause of neuronal death is needed to properly choose the most effective therapeutic approach (single drug or multiple drug regimens). Therapeutic assessment should be conducted in patients with certain diagnosis. This apparently evident prerequisite does not however appear to be easy to satisfy as has been demonstrated by anatomoclinical series in PSP and MSA, and even more so in CBD. Use of international criteria does not alleviate the difficulty. Satisfactory criteria of efficacy remain to be identified. Assuming that such trials would be conclusive, there remains the question of how to implement neuroprotection in routine practice. The difficulties encountered are well known: late intervention after development of the disease in sporadic cases, ethical issues concerning preclinical screening in familial forms of the disease or in patients exposed to an exotoxin. PMID- 12773895 TI - [Angelman syndrome: clinical and genetic model]. AB - Angelman syndrome is a neurogenetic disorder characterized by developmental delay, absence of speech, motor impairment, epilepsy and a peculiar behavioural phenotype associated with abnormalities of chromosome 15q11-13. Precise diagnosis carries clinical and genetic counseling implications. However, many clinicians still seem unfamiliar with it despite the severity and typical aspects of presentation. Beyond individual situations, Angelman syndrome can serve as a model opening broad questioning of genetic and epigenetic influences in neurology, as well as of several concepts such as psychomotor development, cerebral palsy, behavioural phenotypes and epileptic syndromes. Recent advances in molecular biology and animal models of the syndrome provide new data which must be included in our interpretation of Angelman syndrome. PMID- 12773896 TI - [Neurological manifestations of fish, mollusk and shellfish poisoning]. AB - Sea-food poisoning is observed in several areas of the world. Intoxication results from ingestion of fresh fish, mollusks, or shellfish contaminated by toxins produced by microorganisms (dinoflagellates). Neurological manifestations are sometimes associated with signs and may be life-threatening. We describe here the principle toxins, their geographic distribution, clinical manifestations, therapeutic management, and possible prevention measures. PMID- 12773897 TI - [Clinical and electrophysiological features of "cortical tremor"]. AB - The purpose of this paper is to draw attention to one particular variant of tremor named "cortical tremor," which corresponds to rhythmic cortical myoclonus whose principal differential diagnosis is essential tremor. Diagnosis may be established with electrophysiological explorations (electromyography, accelerometry, electroencelography with back averaging, long latency reflexes, giant somesthesic evoked potentials). Clinical and electrophysiological features of 15 patients are reported and compared to cases previously published. Association with generalized or partial seizures is possible. Other cases in the family are often mentioned with a dominant autosomal transmission. The therapeutic approach is based on the use of antiepileptic drugs. PMID- 12773898 TI - [Neuropsychological and scintigraphic aspects of frontotemporal dementia preceding amyotrophic lateral sclerosis]. AB - Between 1993 and 2001, we observed fifteen patients (ten men and five women, mean age 63 years) with frontotemporal dementia (FTD) which preceded signs of amyotrophic lateral sclerosis (ALS) which developed 21 months later. Mean disease duration in the fourteen deceased patients was 38 months. FTD associated with ALS is characterized by rapid course, predominance of disinhibited forms (orbito basal), presence of aphasia with neologisms, and semantic memory disorders. Performed in all patients, single-photon emission computed tomography demonstrated a bifrontal pattern of low uptake, sometimes associated with low uptake in the anterior temporal region. In one patient, neuropathology revealed neuron atrophy and loss in the frontotemporal region, the anterior horns, and the hypoglossal nucleus. Ubiquitin-positive inclusions were visible in the dentate gyrus of the hippocampus and in the anterior horns. The dementia/ALS association is classically described is uncommon. It belongs to the FTD group since the Lund and Manchester consensus. Approximately 15 p.100 of patient with FTD can be expected to develop ALS. About 250 cases have been reported in the literature, half of them in the Pacific area where the incidence of ALS is high (55/100,000 inhabitants versus 1/100,000 in the rest of the world). Intermediary forms of FTD, semantic dementia, and progressive non-fluent aphasia are discussed since several cases of non-fluent progressive aphasia associated with ALS are reported in the literature. The links between these two degenerative diseases are discussed. PMID- 12773899 TI - [Management of stroke in France. Results of 3 national surveys]. AB - Three surveys were performed in France, in March 1999, to analyze the management of acute stroke patients. Three hundred forty five Emergency departments, 93 departments of Neurology and 258 Rehabilitation units participated. Fifty per cent of patients with stroke arrive at the emergency department within 3 hours of symptom onset. Only 40 p. cent of the patients are admitted in a neurological department and 5 p. cent in an acute stroke unit. The mean length of acute hospital stay is often very long (more than three weeks) because of the delay to transfer to a rehabilitation unit. This study shows that acute stroke management is herogeneous and not structured in France. PMID- 12773900 TI - [A descriptive analysis of naratriptan use among migraineurs in ambulatory medicine]. AB - The objective of this study was to provide an epidemiological description of naratriptan use in ambulatory medicine. 1695 patients were recruited by 384 primary care physicians and 111 neurologists, and followed for 12 weeks. Physicians had to document the migraine history, and to report symptoms and health care in a structured case report form. Patients were to document each episode of migraine (EM) in a diary. At baseline, 45 p.cent of the patients reported their migraine treatment as unsatisfactory. Ninety-eight percent of included patients were migraineurs according to criteria of the International Headache Society (IHS), including migrainous disorders. Ninety-two percent of naratriptan prescriptions were established in the second intention in patients with migraine, according to the IHS classification, including migrainous disorders. A total of 79 p.cent of patients had complied with the good practices for all EMs. More appropriate health education strategies should target the small group of patients who over-use naratriptan, and patients with aura. However, this study shows that naratriptan tends to be correctly prescribed by physicians, and used by patients with acute migraine. PMID- 12773901 TI - [Bilateral ageusia after left insular and opercular ischemic stroke]. AB - Gustatory disorders after ischemic stroke is an uncommon event, generally occurring unilaterally, ipsilateral or contralateral to the stroke depending on the site of the lesion. We report a patient who developed bilateral agueusia after a unilateral insular stroke. This 70-year-old right-handed man developed a sudden onset left opercular syndrome in March 1999 with right hemiphyosethesia. Magnetic resonance imaging revealed a left insular and opercular stroke. Anarthria, swallowing and sensorial disorders regressed in a few weeks, but taste disturbances led to a 6 kg weight loss. After six months, the symptoms disappeared. To our knowledge, 32 cases of taste disorders after stroke have been reported. For most of the cases, the agueusia was unilateral. Our case illustrated the predominance of the left cortical hemisphere for taste perception in the right-handed subject. PMID- 12773902 TI - [Temporal lobe epilepsy and complete bilamination of the granule cell layer of the dentate gyrus]. AB - A bilamination involving the whole dentate stratum granulosum associated with a hippocampal sclerosis is reported. This morphological abnormality could be an unusual aspect of granule cell dispersion, plastic change induced by an early post-natal injury, or the the result from a neuronal migration disorder during the embryonic period. Whatever its origin, this bilamination is an abnormality of the hippocampal development which continues during the first years of life. PMID- 12773903 TI - [Livedo-like dermatitis (Nicolau's syndrome) after injection of Copolymer-1 (Glatiramer acetate)]. AB - We report the first case of a 33-year-old woman with multiple sclerosis, who developed a livedo-like dermatitis after injection of Copolymere-1. This disease is characterized by the development of acute violent pain during or immediately after injection, and a livedo-like plaque followed by necrosis corresponding to an arterial ischemia by vasospasm or thrombosis. Early treatment with vasoactive and anticoagulation agents is required. Surgery may be necessary. PMID- 12773904 TI - [A new case report of spinal cord compression secondary to beta-thalassemia]. AB - Extramedullary hematopoiesis is an unusual cause of spinal cord compression. We report the case of a 39-year-old man who presented symptoms of progressive dorsal spinal cord compression. Hemogramm showed erythrocytosis with microcytosis. Hemoglobin electrophoresis revealed heterozygous beta-thalassemia. Diagnosis of extramedullary hematopoiesis was based on magnetic resonance imaging findings. Radiological and therapeutic options in this rare condition are briefly discussed. PMID- 12773905 TI - [Subacute myelitis revealed by human immunodeficiency virus infection]. AB - A 35 year-old heterosexual man had a six months history of cervical myelitis with progressive paraplegia, leg weakness and paresthesia of the four extremities. Spinal cord MRI showed a high T2 signal intramedullary lesion wide from the bulbo medullary junction to D4. Post gadolinium T1 sequence revealed an enhancement in front of C3-C4 vertebrae. VIH serology was positive. Corticosteroid treatment achieved a marked improvement. In addition to vacuolar myelopathy, well-known at the advanced stages of the HIV infection (AIDS), myelitis and clinical pictures simulating multiple sclerosis were described during early stages of the infection. These inflammatory lesions of the central nervous system and sometimes of the peripheral nervous system seems to be related to the immune response dysfunction induced by the VIH. PMID- 12773906 TI - [Castleman's disease located in the central nervous system]. AB - A 26-year-old woman complained of trijeminal nevralgy and ocular symptoms revealing a paracavernal tumor which had progressed for three months. Histopathological analysis after partial resection led to the diagnosis of Castleman's disease. Six months later, the patient was considered cured after focal adjuvant radiotherapy. Castleman's disease is a lymphoproliferative disorder. Solitary intracranial involvement is unusual. Unlike multifocal disease, localized Castleman's disease has an excellent prognosis. PMID- 12773907 TI - [Contrast uptake after spinal tap: myth or reality?]. PMID- 12773908 TI - [Multinodular presentation of multiple sclerosis]. PMID- 12773909 TI - [Management of a parkinsonian patient treated by stimulation of the subthalamic nuclei]. PMID- 12773910 TI - [Ophthalmic manifestations of Horton disease]. PMID- 12773912 TI - [Pathophysiology and diagnosis of polycystic ovary syndrome]. PMID- 12773913 TI - [Epidemiology and genetics of polycystic ovary syndrome: recent date]. PMID- 12773914 TI - [Contribution of ultrasonography for the diagnosis of polycystic ovaries]. PMID- 12773915 TI - [Treatment of PCOS without IVF: weight loss, insulin-sensitizing agents]. PMID- 12773916 TI - [Ovary stimulation without IVF in polycystic ovary syndrome]. PMID- 12773917 TI - [Ovary stimulation with IVF in women with polycytstic ovary syndrome]. PMID- 12773918 TI - [Overstimulation of the ovary: prevention and treatment]. PMID- 12773919 TI - [Reasons for surgical treatment of polycystic ovary syndrome]. PMID- 12773920 TI - [Reasons against surgical treatment of polycystic ovary syndrome]. PMID- 12773921 TI - [In vitro maturation: clinical and biological aspects]. PMID- 12773923 TI - [Magnetic resonance imaging applications in obstetrics]. AB - PURPOSE: To review the main indications and results of magnetic resonance imaging in the pregnant women. MATERIAL AND METHOD: We reviewed MRI practice during the pregnancy based on our own experience in a prenatal diagnostic center and data in the literature. Rapid improvement in MRI technology has allowed more extensive use, giving a good contrast-to-noise ratio and multiplanar imaging. RESULTS: Although ultrasound provides primary screening information, final diagnosis may require further investigations. MRI, to be performed in the second and third trimester, is the non-invasive second line tool of choice in this context. The most widespread indications are for brain disease: search for a cause of ventriculomegaly or biometric abnormality, confirmation of a malformative or acquired lesion. Progressively, indications were widened to head and neck, thorax, abdomen and pelvis areas. Moreover, systematic indications include previous fetal pathology or the pregnancy context. Other MRI indications have been suggested: placental malposition, pelvimetry and maternal genito-urinary tract. CONCLUSION: MRI is becoming the natural and necessary second line imaging technique, with increasing indications. It must be kept in mind however that all pathological conditions cannot be depicted by these morphological studies. PMID- 12773924 TI - [Dating biometry with crown-rump length, precision of a routine practice]. AB - OBJECTIVE: The goal of this study was to determine the accuracy of an everyday practice for assessing gestational age by ultrasound and redefine the correction of gestational age policy. MATERIALS AND METHODS: This study used first trimester measurements taken during a three-year period. We considered all births from pregnancies that began by an in vitro fertilization procedure. We examined 143 consecutive files containing 257 measurements made by 72 different operators. We applied two reference curves to calculate the date the pregnancy began and the centiles of the prediction interval for +/- 7, +/- 5 and +/- 3 days. RESULTS: The prediction intervals for +/-7, +/-5, +/-3 days excluded 2%, 6%, and 25% respectively of the embryos from one of the two reference curves. These intervals were 1%, 5% and 20%, were better for the other curve. CONCLUSION: Correction of gestational age has to take into consideration variations in the embryo length. We correct the gestational age only if the difference with ultrasound assessment is more than one week. PMID- 12773925 TI - [Evaluation of maternal morbidity after drug-induced termination of pregnancy (TOP) after 12 gestation weeks]. AB - OBJECTIVE: The number of TOP for medical indications has increased regularly over the last ten years. At the same time, the methods used for TOP have evolved. The purpose of this study was to assess the frequency of complications after drug induced termination of pregnancy in order to determine whether using this method for interrupting pregnancy during the second or third trimester adds further danger for the mother in terms of early severe risk (uterine rupture, hysterectomy) or less severe long-term risk (infection). MATERIAL AND METHODS: This metaanalysis included all articles devoted to pregnancy interruption from 12 gestation weeks retrieved from the Medline database and published between 1989 and 1999 in the United States, Canada, Australia, New Zealand, or the European Union. After excluding articles that included in utero death (n=8), isolated case reports and series involving a high-risk of maternal somatic complications (n=16), and surgical methods for pregnancy termination (n=4), we retained 23 articles for analysis. These articles had included 58,891 drug-induced terminations of pregnancy. For each article, we recorded the following complications: bleeding requiring transfusion, uterine rupture, ovular or placentar retention, and infection. A classical homogenicity test was performed for each type of complication. When this test was not significant, a mean rate, weighing by size of the study, was calculated. RESULTS: One study reported maternal deaths (3/143000). The weighted mean rate for late retention (>24 hr) was 1.5 [CI95: 1.1%-1.9%]. For infections, the rates were very variable between studies (from 0.7% to 3.6% with one study reporting 8%). For bleeding with transfusion, the weighted mean rate was 0.7% [CI95: 0.5%-0.9%]. This rate was significantly higher than the rate observed in 1999 in France after delivery excepting medically terminated pregnancy (p<10(-3)) but probably is a reflection of the variable transfusion practices during the eighties in these different countries. Th rate of uterine rupture after medically terminated pregnancy was 0.1% [CI95: 0.07%-0.17%] and would be higher after delivery (excepting terminations) but not significantly (p=0.07). CONCLUSION: This metaanalysis demonstrates that the risk of severe complications (uterine rupture and bleeding requiring transfusion) are rare but are more prevalent than after delivery except pregnancy termination. The metaanalysis approach is justified due to the low incidence of these severe complications. A prospective multicentric study of the complications using a geographical base would be useful to obtain unbiased data on risk level. A risk analysis by gestational age, maternal age, parity, and product used would thus be possible, as would long-term monitoring of maternal outcome. PMID- 12773926 TI - [Therapeutics indications and prognosis of eclampsia at Dakar University Teaching Hospital]. AB - OBJECTIVE: To study the factors of risk of complications and evaluate the quality of medical and obstetrical management of eclampsia. METHODOLOGY: We performed a retrospective analysis of all cases of eclampsia recorded during the ante- or per partum period from January 1, 2000 to December 31, 2001 at the gynecological and obstetrical clinic at Dakar University Teaching Hospital. RESULTS: Eclampsia had concerned 78 patients, with an incidence of 8 per 1000 childbirth. The epidemiological profile was primipara (68%), young (average age: 20 years), 36 weeks pregnancy or more (60,3%), evacuation in a state of post critical coma (74.3%), on average after 2 seizures, without medical assistance (64%) with an average diastolic blood pressure of 100 mm Hg. Examinations with strong prognosis value like blood count, creatininemia, coagulation, hepatic transaminases and uricemia were not available in emergency (only 24% of all cases). Medicines prescribed included, nicardipine (63%) and magnesia sulfate (53%); this medical treatment was satisfactory in 34.6% of the cases. Cesarean section was performed in 50%. The prognosis was marked by 17.9% maternal mortality and a perinatal mortality of 359 per 1000 births. The principal risk factors of maternal and perinatal complications were early-onset eclampsia, large number of seizures and late obstetrical treatment. CONCLUSION: Prehospital treatment and availability of early cesarean section must be better organized to improve the prognosis of eclampsia. PMID- 12773927 TI - [Norplant contraceptive implant]. AB - OBJECTIVE: Norplant is a long lasting contraceptive method which protect again pregnancy for five years. We studied the side effects and the variations related to weight and blood pressure. MATERIAL AND METHODS: The study was carried out in three family planning clinics in Dakar with 450 women who have had their implants for at least six months. Data collection was based on patient interviews and review of medical files. RESULTS: The side effects reported were mainly menstrual problems with 54.7% of the women reporting menstrual spotting. The other non menstruel side effects were headaches (64.2%) and dizziness (59.1%). The side effects at the insertion site were pruritus (16%) and pain (3.1%). Regarding weight and blood pressure, an increase in weight was observed up to three years of Norplant use and a decrease to the initial weight was seen during the fifth year of use. No variation was noted regarding blood pressure. CONCLUSION: Norplant is a contraceptive method widely used in Senegal. Most of the side effects involve menstrual problems. A slight increase of weight was observed in general with no variation in blood pressure. PMID- 12773928 TI - [Contribution of Doppler ultrasound to the antenatal diagnosis of persistent right umbilical vein: case report]. AB - We report a case of an isolated persistent right umbilical vein which was diagnosed on an ultrasound examination performed in the third trimester of pregnancy. This was made possible by the contribution of color flow Doppler and real-time ultrasound. Having isolated the anomaly no further fetal monitoring was instituted. Delivery was at term and the infant is doing well. Persistent right umbilical vein results from an anomaly of organogenesis. It represents the main intra abdominal anomaly of this vein. Its incidence is estimated at approximately 1 case for 500 antenatal ultrasound examinations. Ultrasonographic diagnosis is relatively easy and can be performed near the end of the first trimester. After insertion in the abdominal wall, the umbilical vein passes round the gallbladder from the right before connecting to the portal vein in the intra-hepatic form; right atrium, inferior vena cava, superior vena cava or iliac vein in extra hepatic form. Diagnosis is affirmed by color flow Doppler, which emphasizes a continuity of blood flow between the extra and intra abdominal segment of the umbilical vein, and real-time ultrasound, which shows a venous blood flow. Prognosis depends essentially on the existence of congenital malformations, which are observed in 10 to 25% of cases. When isolated, persistent right umbilical vein is generally associated with favorable outcome and must be considered as a normal fetal anatomical variant. PMID- 12773929 TI - [Management of ectopic pregnancies complicating in vitro fertilization: a remarkable case of bilateral ectopic pregnancy with independent courses of the pregnancies]. AB - OBJECTIVE: To define management rules for ectopic pregnancy complicating in vitro fertilization. PATIENT AND METHOD: We describe a case of bilateral ectopic pregnancy complicating in vitro fertilization which led to two separate laparoscopic procedures. RESULTS: A first laparoscopy was required to perform salpingectomy for ectopic pregnancy. A second laparoscopy was later required for a second salpingectomy for a second ectopic pregnancy complicating the same in vitro fertilization. DISCUSSION AND CONCLUSION: Pre-existent tubal lesions are frequently observed in women who develop ectopic pregnancies after in vitro fertilization. If serum beta-hCG levels and pelvic ultrasound results are not taken into consideration with sufficient care, delayed diagnosis may lead to the unsuitable management. Women with multiple embryonic reimplantations must be monitored rigorously. Salpingectomy is the best treatment option for this type of complication. PMID- 12773930 TI - [Virilism during pregnancy]. AB - We discuss the causes of virilism in pregnancy and the impact of hyperandrogeny on the female foetus. We report a case of virilism in a 28-year-old, gravida 1, para 1 patient with normal pregnancy and review the literature. After conception, the patient had been well until the 18th week of gestation, when she developed signs of virilization; her serum testosterone was markedly elevated. She delivered of a normal male infant at term. After delivery, signs of virilization regressed with normalization of testosterone level but a wide mass of the left ovary that persisted for 8 months. The histological study of the tumor showed luteinized thecoma. Luteomas and hyper-reactio luteinalis were the principal causes of virilism in pregnancy, thecomas are rare. PMID- 12773931 TI - [French and European trainees in OB/GYN are opposed to a specific and exclusive French training program in medical gynecology]. PMID- 12773934 TI - Endocrinology of menopause and "andropause". Proceedings of the 46th International Meeting of Clinical Endocrinology Henri-Pierre Klotz. Paris, France, 2003. PMID- 12773933 TI - [Management of acromegaly in 2003]. PMID- 12773935 TI - The endocrinology of gonadal involution: menopause and andropause. AB - Most aging individuals die from atherosclerosis, cancer or dementia. In the oldest old also loss of muscle strength resulting in frailty becomes the limiting factor for an individual's chances of living an independent life until death. Two hormonal changes mark the aging process in man. In women an acute drop in estrogen production by the ovaries around the age of 50 initiates a symptom complex called menopause. In men a more subtle drop of testosterone bioactivity from 40 yrs onwards might be accompanied by more difficulty to recognize symptomatology (andropause). Hormone replacement strategies in elderly women and males with estrogens or androgens, respectively, has some clear advantages, but is currently controversial, because of the occurrence of adverse effects. PMID- 12773936 TI - Oocyte apoptosis: prevention strategies, and implications for female aging and the menopause. PMID- 12773937 TI - Role of the first waves of growing follicles in rat ovarian maturation. PMID- 12773939 TI - Mechanisms of premature ovarian failure. AB - The median age at menopause in Western populations of women is approximately 51 years. While very late (i.e., after 54 years) menopause is exceedingly uncommon, a sizeable minority of women experience cessation of ovarian function at or prior to age 45. By convention, menopause that occurs at ages 40-45 is considered "early" and occurs in about 5% of women. Premature ovarian failure (POF) is reserved for the approximately 1% of women who experience hypergonadotropic amenorrhea prior to age 40 years. POF represents the end stage of a variety of disorders that result in the loss of ovarian follicles. Depending upon the age at diagnosis, the probability of a genetic, autoimmune, or idiopathic cause will be more or less likely. Two functioning X chromosomes appear necessary for normal ovarian function. The most obvious genetic cause of POF is Turner Syndrome, in which a complete or near-complete loss of the second X chromosome occurs. Turner Syndrome typically results in the most severe and irreversible POF, often clinically evident prior to menarche. Typically, in Turner Syndrome, menopause precedes menarche, and there is no evidence of ovarian function. However, cases with multiple tissues diagnosed as 45, X have been reported to result in ovarian function and even pregnancy. It is likely that mitigating factors, perhaps autosomal, can modify this most severe and irreversible cause of ovarian failure. Lesser degrees of ovarian failure have also been attributed to partial X chromosome deletions and milder degrees of X chromosome mosaicism. Fragile X syndrome is another example of mild POF that can be linked to disorders of the X chromosome. Other genetic defects are believed to cause POF, yet their prevalence has been difficult to determine. The localization of the gene for the blepharophimosis/ptosis/POF Syndrome has been recently reported, yet this finding has not been seen commonly in POF. Other genetic syndromes including POF await elucidation. Many transgenic "knock-out" animals have been created with deficient ovarian function. Most interesting along these lines is the heterozygous FSH receptor knock-out, which exhibits a reduced follicle reserve and early ovarian depletion. Application of this knowledge and translation of these transgenic experiments into elucidation of clinical disease has been difficult, but represents an area of tremendous potential progress in the understanding of the pathogenesis of POF. Another approach to the genetics of POF has been to examine the genome of affected and unaffected individuals. The genetics appear to differ greatly depending upon the timing of the expression of the POF. For example, women with early menopause are more likely to possess the PVUII polymorphic allele for estrogen receptor alpha. Whether or not this polymorph is more common in women with earlier menopause, i.e., POF, is unclear. Pedigree data indicate that early menopause and premature menopause sort similarly within families. The only difference between women with true POF and those with early menopause may be in the timing of the expression of the syndrome, and not in the genetics. Population genetic approaches analyzing affected and unaffected individuals are underway in several research centers and represent another area of progress. Immune and other, idiopathic causes of POF await further clarification. It is clear that this is an area of great research potential. Understanding how ovaries fail may assist women with this disorder by facilitating the development of novel therapies. Additionally, such information will provide important clues about optimizing ovarian function in individuals without POF who are seeking extension of their reproductive life spans or fertility enhancement by other means. PMID- 12773938 TI - Age-related analysis of inhibin A and B relative to the intercycle monotropic FSH rise in normal ovulatory women. PMID- 12773940 TI - [Early menopause and hypothalamo-hypophyseal radiographic anomalies]. PMID- 12773941 TI - Post menopausal ovarian steroidogenesis. PMID- 12773942 TI - Extragonadal synthesis of sex steroids: intracrinology. AB - All androgens and estrogens in post-menopausal women are synthesized locally in peripheral target tissues by tissue-specific steroidogenic enzymes according to the intracrine process. The importance of the intracrine or peripheral formation of sex steroids is illustrated by the success of aromatase inhibitors and antiestrogens in the prevention and treatment of breast cancer in post-menopausal women. On the other hand, a large series of problems are associated with the deficiency of sex steroids accompanying menopause and the decreased secretion of DHEA, osteoporosis being the best defined example. In this context, an important observation is that women at menopause are not only deprived from ovarian estrogens but are also lacking androgens due to the marked decrease of DHEA. In order to achieve a more physiological and tissue-specific hormone replacement therapy, DHEA, in combination with a SERM (selective estrogen receptor modulator) having pure antiestrogenic activity in the mammary gland and uterus could possibly meet the most important needs of women at menopause, namely control of hot flushes and, most importantly, prevent breast cancer, uterine cancer, ovarian cancer and osteoporosis with a potential improvement of cognitive functions and memory and prevention of Alzheimer's disease. PMID- 12773943 TI - A round table to avoid going round in circles and missing the testosterone decline phenomenon in aging males. PMID- 12773944 TI - Diagnosis of partial androgen deficiency in the aging male. AB - From the age of 30 yrs on, the (free) testosterone [(F)T] levels decrease continuously with age, mean total T level at age 70 yrs, being only 2/3rds of the mean level at age 25 yrs, whereas mean FT level is only 40% of the mean level in young adults. However, inter-individual variations are wide and whereas at least 30% of men over age 70 yrs have clearly subnormal (F)T levels, 20% have still levels in the upper range of values in young men. Aging in males is accompanied by a series of signs and symptoms, reminiscent of androgen deficiency in young adults, such as decrease in muscle mass and strength, increase in abdominal, mainly visceral, fat with insulin resistance and atherogenic lipid profile, decrease in libido and sexual hair, osteopenia, decrease in cognitive performances, insomnia, excessive sweating and decrease in general well-being, and it is tempting to relate these symptoms to the age associated decrease in androgen levels, the more that often a significant, albeit generally weak, correlation of these symptoms with the (F)T levels is observed. Moreover, the preliminary data suggest favourable effects of androgen supplementation in the elderly. The decrease in (F)T levels is, however, only one of the factors responsible for the signs and symptoms of the aging male which have a multifactorial origin. Hence, the diagnosis of androgen deficiency in elderly men should be based on both the clinical symptomatology and the biochemistry, i.e. decreased (F)T levels, the latter being defined, more or less arbitrary, as levels below the lowest 1% of levels in young healthy males. PMID- 12773945 TI - Assays for testosterone in plasma: untested assays for an unproven syndrome. PMID- 12773946 TI - Bioavailable testosterone as index of androgenicity (from 1973 to 2003). PMID- 12773948 TI - Significance of oestrogens in male (patho)physiology. AB - Traditionally conceptualized as 'female hormones', oestrogens appear to have significant effects in the male biological system. Favorable effects have been noted on bone, brain and cardiovascular physiology while a potential role in the prostate pathology of the aging male has been seriously suspected. Oestrogens in male are predominantly the products of peripheral aromatization of testicular and adrenal androgens. While the testicular and adrenal production of androgens declines with aging, levels of total plasma oestradiol do not decline. This is to be ascribed to the common increase in fat mass with aging (the substrate of peripheral aromatization) and an increased aromatase activity with aging. But free or bioavailable oestrogens may decline due to an increase in sex hormone binding globulin. Oestrogens produce significant beneficial effects on skeletal growth and bone maturation. In old age oestrogens are better predictors of bone fractures than androgens. Oestrogens exert effects on the brain: on cognitive function, co-ordination of movement, pain and affective state, and are maybe protective of Alzheimer's disease. Oestrogen effects on the cardiovascular system include those on lipid profiles, fat distribution, endocrine/paracrine factors produced by the vascular wall (such as endothelins, nitric oxide), blood platelets, inflammatory factors and coagulation. The potentially adverse effects of oestrogens on the prostate may be due to a shift in the oestrogen / androgen ratio with aging. Sources of estrogens in men are endogenous androgens, or in case of androgen deficiency, exogenous androgens. Dietary phytoestrogens or selective estrogen receptor modulators, as drugs, may be significant as well. PMID- 12773947 TI - [Contribution of bioavailable testosterone assay for the diagnosis of androgen deficiency in elderly men]. AB - With age, some men develop symptoms resembling hypogonadism. Several cross sectional and longitudinal studies have shown a decrease in testosterone levels with ageing in men. This finding has equally been observed in elderly men in good health. Testosterone levels decline progressively as of the thirties, at a rate which remains constant throughout life. While total testosterone levels decrease, sex hormone binding globulin (SHBG) levels on the contrary increase with age, with the result that the levels of free and non-SHBG-bound testosterone (corresponding to the fraction which is bioavailable to target cells) decrease more abruptly than that of total testosterone. Higher LH levels, decreased testosterone response to hCG and less Leydig cells all indicate that ageing induces partial testicular failure. However, the gonadotropic function is also affected in ageing. The hypothalamus-pituitary becomes more sensitive to gonad steroid feedback, LH pulse amplitude decreases, and the LH response to GnRH is blunted compared to the situation in young men. Thus LH level is not a valid index of androgen deficiency in elderly males. None of the androgen-dependent functions (libido, erection, sense of well-being, muscle mass and strength, fat mass, bone mass, erythropoiesis, etc.) are under exclusively androgen control, and there is no elderly male symptom which is completely specific to androgen deficiency. Thus, in elderly men, when clinical symptoms might indicate androgen deficiency, biological confirmation is needed. An assay which is independent of SHBG fluctuations is mandatory. Bioavailable testosterone assay by ammonium sulfate precipitation seems to us to be the optimum method for diagnosing androgen deficiency: it gives a reliable measurement for the testosterone fraction available to target cells, is adapted to clinical practice, and provides results that can be directly compared with current reference values for healthy young men. PMID- 12773949 TI - Partial Androgen Deficiency of Aging Male (P.A.D.A.M.) might in part be due to excessive organochlonide pesticide (OC) impregnation. PMID- 12773950 TI - [Estrogens, androgens, and osteoporosis in men]. AB - In males, estrogens are synthetized in peripheral tissue by aromatization of testosterone. Recent data show that males either with a non functional estrogen receptor or unable to synthetize estrogen have osteoporosis although they have normal circulating levels of androgens. Several transversal studies show that, in aged males, estrogens are better correlated to bone density than androgens. Free estradiol and free androgen levels are both decreased during ageing between 50 and 80 years old. Estrogen rather than androgen deficiency is responsible of increased bone resorption in aged males. So far there is no treatment without side effect for the treatment of estrogen deficiency in males. PMID- 12773951 TI - [Osteoporosis in the elderly man]. AB - Elderly men are at substantial risk for fracture. Morbidity after osteoporotic fractures appears to be more serious and mortality more common in men than in women. Risk factors for osteoporotic fractures in men appear to be qualitatively similar to those in women. Low bone mineral density (BMD) is an important risk factor for fracture in men; however, further clarification of the relationship between BMD, bone geometry and fracture risk is needed. Our understanding of the mechanisms underlying senile bone loss and the pathogenesis of senile osteoporosis in men remains fragmentary with, in particular, the need for further clarification regarding the precise impact of hormonal status in elderly men on skeletal homeostasis. Nevertheless, the available evidence indicates a role for both testosterone and estrogens in the regulation of bone metabolism in elderly men. Recommendations concerning prevention and treatment of senile osteoporosis in men should focus on the minimization of known risk factors for bone loss and falls. Testosterone treatment may be useful only in those men with initially low serum testosterone. As to other pharmacological treatment modalities, prospective trials specifically in elderly men, and preferably with fracture incidence as the primary clinical endpoint, are required. PMID- 12773952 TI - [Consequences of estrogen and androgen deficiency on fat metabolism]. PMID- 12773953 TI - Sex steroids and sleep: sleep disturbances in menopause. PMID- 12773954 TI - Relationship between REM sleep and testosterone secretion in older men. PMID- 12773955 TI - Cognitive functions and sex steroids. AB - In humans, levels of adrenal (DHEA(S)) and gonadal (estradiol, testosterone) sex steroids decline with age. Studies in rodents have demonstrated that these hormones can have neuro-excitatory and neuro-protective effects in the central nervous system (CNS). Behavioural studies repeatedly have reported enhanced memory performance of rats and mice after acute or sub-chronic treatment with sex steroids. The current review summarizes human studies on this topic conducted by the author as well as other groups. Epidemiological as well as experimental studies have in general shown that estradiol replacement improves cognition, especially verbal memory in menopausal women. Similarly positive effects of testosterone replacement in older men have been reported in several, but not all studies. Cognition enhancing effects of DHEA replacement in older healthy humans in contrast could not be demonstrated with short (weeks) or prolonged (months) treatment regimes. Even though most results support the notion that estradiol in women and testosterone in men can enhance cognition in older healthy humans, more research is needed before recommendations for the clinical practice can be made. PMID- 12773956 TI - [Testosterone and depression in men aged over 50 years. Andropause and psychopathology: minimal systemic work-up]. AB - A reduced feeling of well being with unusual anxiety and irritability, nervousness, mood swings and a depressive state are often mentioned as the psychological symptoms of the age-related hypogonadism. However, psychological aspect of andropause has not yet been specifically studied and most data on psychological symptoms come from researchers' clinical impressions rather than from systematic studies. Therefore, it seems premature to assign them to the age associated decline in testosterone levels. The implication of testosterone in psychological state has yielded mixed results. Among elderly men, lower testosterone levels were associated with depressive or dysthymic symptoms. Moreover, lower testosterone levels were reported in men with depression, independently of age. In contrast, some studies did not observe any significant difference in testosterone levels between depressed men and controls. Furthermore, several studies have suggested that testosterone replacement improved mood in hypogonadal men, but others did not, as in studies on eugonadal men. Several researchers have also suggested the potential use of testosterone as an antidepressant or adjuvant to current treatments in depressed hypogonadal men. The relationship between andropause and psychological symptoms such as depression is far from clear. Andropause may be associated with "minor depressive symptoms" that are not considered as pathological. Psychological manifestations do not appear specific to andropause and probably have a multifactorial origin. PMID- 12773957 TI - Metabolic effects of oestrogens: impact of the route of administration. AB - To investigate whether oestrogen modulates GH secretion and action in adult life, we studied the impact of oestrogen replacement on circulating GH and IGF-I levels in post-menopausal women. Since the liver is the major source of circulating IGF I and the oral route of oestrogen delivery causes non-physiologic effects on hepatic proteins, we compared the effects of oral and transdermal route of delivery. Oral ethinyl oestradiol administration resulted in a significant fall in mean IGF-I levels and a 3-fold increase in mean 24h GH. Transdermal administration of 17beta oestradiol resulted in a slight increase in serum IGF-I but no change in mean 24h GH levels. To determine whether differences in oestrogen type rather than in the route of delivery caused the different effects on the GH/IGF-I axis, we compared the effects of three oral oestrogen formulations. Ethinyl oestradiol, conjugated equine oestrogen and oestradiol valerate each induced a fall in IGF-I and a rise of mean 24h GH levels in post menopausal women. To determine the metabolic significance of oestrogen-induced changes on GH/ IGF-I, we compared the effects of 24 weeks each of oral and transdermal oestrogen on energy metabolism and body composition in 18 post menopausal women in an open-label randomised cross-over study. When compared to the transdermal route, oral oestrogen reduced lipid oxidation, increased fat mass and reduced lean body mass. Oestrogen causes distinct, route dependent effects on the somatotrophic axis. The dissociation of the GH/IGF-I axis by the oral route is likely to arise from impaired hepatic IGF-I production which causes increased GH secretion through reduced feedback inhibition. The route of oestrogen therapy confers divergent effects on substrate oxidation and body composition. The suppression of lipid oxidation during oral oestrogen therapy may increase fat mass while the fall in IGF-I may lead to a loss of lean body mass. The route dependent changes in body composition observed during oestrogen replacement therapy may have important implications for post-menopausal health and oestrogen use in general. PMID- 12773958 TI - Progestins in hormonal replacement therapy (HRT): new molecules, risks and benefits. PMID- 12773959 TI - The "climacterium virile": from Lagueur's hesitations to our legitimate hedging in 2003. PMID- 12773960 TI - [SERM: mechanisms of action and therapeutic indications]. PMID- 12773962 TI - The problem of late allograft loss in kidney transplantation. AB - The 2 principal factors implicated in late kidney allograft failure are chronic rejection (also called chronic allograft nephropathy) and death of the patient with a functioning graft (mainly from cardiovascular causes). Despite lifelong immunosuppression of the recipient, immunological responses remain the leading factor in the pathogenesis of chronic rejection and both cellular and humoral immune mechanisms have been shown to play important roles. In this review, we highlight the relevance of humoral mechanisms of rejection to the pathogenesis of late allograft loss. Non immunological factors, such as donor organ quality, initial ischemic injury, calcineurin inhibitor (CNI) toxicity, hypertension, and hyperlipidemia, also contribute to progressive chronic allograft injury, but will not be reviewed in detail here. Possible strategies to stabilize or improve allograft function in patients with already established "chronic rejection/chronic allograft nephropathy" (CR/CAN) are the addition of mycophenolate mofetil (or sirolimus) with or without a reduction of cyclosporine dosage, or conversion from cyclosporine to tacrolimus. However, prospective randomized clinical trials are needed to test the efficacy of these strategies. A major current challenge for transplant physicians is to develop regimens that prevent CR/CAN, since, once established, the process typically progresses inexorably to renal allograft loss in most recipients. Evidence is now accumulating that new immunosuppressive regimens must control not only T cell but also B cell responses (i.e. limit antidonor antibody production) in order to prevent CR/CAN and improve long-term allograft survival. PMID- 12773961 TI - New tissue-selective androgens: perspectives in the treatment of androgen deficits. PMID- 12773963 TI - Long-term renal transplantation. Recent successes and new problems. AB - There has been continued improvement in preventing early biopsy-proved acute rejections and increasing 1-year allograft success rates, and, after some delay, the conditional half-life of grafts. Beneath these impressive achievements are several troubling concerns: unrecognized subclinical rejections; the emergence of acute and chronic humoral rejection; the different effect of certain acute rejections on the development of chronic rejection; the possibility that the current improvements in reducing early acute rejection may not be translated into longer half-life for the graft unless important adjunctive therapy is included. The use of cyclosporine (CSA) has been reshaped by lower dosing, conversion and avoidance protocols, C2 blood level monitoring, availability of generics, and application to non-transplant immunologically-mediated renal diseases. The enigma of chronic allograft nephropathy (CAN) unfortunately still remains, but recent studies on the effects of hypomagnesemia are provocative. The actual cohort of truly long-term renal transplant successes is providing remarkable insights into the way such grafts and their recipients achieve 30-year success. More than half of these patients have experienced early acute rejections (even Banff II and III). Somewhat contrary to expectations, these recipients usually do not have subnormal levels of CD4+ and CD8+ lymphocytes. However, they are typically B cell depleted, a condition that may protect them from late humoral rejection. Future directions will likely lead to the inclusion of important adjunctive agents having secondary anti-proliferative and anti-fibrogenic capabilities. Because chronic injury to renal allografts, as well as the dominant complications of renal transplantation such as cardiovascular disease and skin cancers, are woven together by a common theme of excessive proliferative activity, albeit of different cell types, long-term therapy will be directed at both protecting the allograft and the allograft recipient by incorporating as long-term therapy selected anti-proliferative agents that could include inhibitors of the renin angiotensin-aldosterone system, statins, sirolimus, mycophenolic acid and leflunomide. PMID- 12773964 TI - Tacrolimus in renal transplantation. A review of recent data. AB - This overview will cover the main development of the use of Tacrolimus in organ transplantation. It summarizes the large amount of data which have recently appeared in the literature, especially in comparison with Cyclosporine, with recent controlled conversion experience, combination therapies, withdrawal of corticosteroids, the use of Tacrolimus in monotherapy and in paediatric renal transplant patients. Finally cardiovascular risk factors are discussed in the light of the other main risk factors in this particular population of transplant patients. PMID- 12773965 TI - Metabolic aspects of tacrolimus in renal transplantation. Consequences for the choice of an immunosuppressive regimen and for the management of post-transplant diabetes mellitus. AB - The occurrence of post-transplant diabetes mellitus (PTDM) is an important complication after renal transplantation associated with an increased risk of chronic transplant dysfunction and of cardiovascular morbidity and mortality. Both tacrolimus and cyclosporine have been associated with PTDM. In the initial studies, PTDM seemed to occur more often in tacrolimus treated patients than in cyclosporine treated patients. The mechanism by which tacrolimus could cause PTDM was unknown and the relative roles of tacrolimus and corticosteroids, which are often prescribed concomitantly with tacrolimus, were unknown. In several studies we used fasting glucose and insulin levels to assess (peripheral) insulin resistance, and intravenous glucose tolerance tests to assess insulin secretion by the pancreatic b-cells in response to a stimulus (glucose load). Thus, we evaluated the mechanism by which tacrolimus causes glucose metabolic disorders, risk factors for glucose metabolic disorders during tacrolimus treatment, the relative roles of corticosteroids and tacrolimus trough levels in glucose metabolic disorders, and also differences in glucose metabolism between patients using tacrolimus versus patients using cyclosporine. Based on the results of these studies and the available literature, the consequences for the choice of a primary immunosuppressive agent and guidelines for the treatment of PTDM during tacrolimus-based immunosuppression are discussed. PMID- 12773967 TI - New monoclonal antibodies in renal transplantation. AB - A decade of spectacular innovation in maintenance immunosuppression drugs has resulted in dramatic reductions in acute rejection and improvement in short and long term outcome after renal transplantation. However the new drugs continue to lack specificity, many require frequent therapeutic drug monitoring and all are associated with acute and chronic toxicities. The new biologic agents, monoclonal antibodies (chimeric, humanized, and fully human) and receptor-fusion proteins, lack immunogenicity, have long half-life and prolonged biologic effects, require intermittent administration and have minimal toxicity. The specificity and selectively of the targets of the new biologic agents render them less toxic than the oral maintenance drugs and thus could possibly replace the maintenance drugs most associated with long-term toxicity such as the corticosteroids and the calcineurin inhibitors. The recently introduced anti-interleukin 2 receptor (IL 2R) monoclonal antibodies (mAbs) are the prototype of future biologic agents; selective, safe, and inducing prolonged biologic effects. The IL-2R mAbs have been used with a variety of maintenance immunosuppression regimens double therapy with cyclosporine and prednisone, triple therapy with cyclosporine, azathioprine and prednisone and with newer regimens such as cyclosporine or tacrolimus, mycophenolate mofetil (MMF) and prednisone, and most recently with sirolimus, MMF and prednisone. The major thrust of the new biologics in clinical development is to block the co-stimulatory pathway. The first attempt at blockade of the CD40 CD154 with anti-CD154 mAbs was disappointing. Anti-CD 154 therapy was associated with thromboembolic events and acute rejection. Attempts at blocking the CD28-B7s (CD80-CD86) pathway are currently underway with the receptor fusion protein, LEA29Y a second generation CTL4Aig, and humanized mAbs to CD 80 and CD86. LFA1, an adhesion molecule that also participates in the co-stimulatory pathway, has also been targeted with a mAb that binds to the CD11a chain of LFA1. Efalizumab, a humanized anti-CD11a mAb, was shown in a phase I trial to be potentially effective in renal transplantation. A humanized anti-CD45 RB mAb is currently in pre-clinical studies and will likely be tested in a phase I trial of renal transplantation within 1 year. While excellent results with anti-CD45 RB mAbs have been published in experimental transplantation, the mechanism of action of anti-CD45 RB mAbs remains to be determined. Several antibodies that are currently approved for non-transplant indications are currently used in single center clinical trials in renal transplantation including Campath 1 H, a humanized anti CD52 mAb, Rituxamab, an anti-CD20 chimeric mAb, and Infliximab an anti-TNFa chimeric mAb. In addition, several humanized mutagenized anti-CD3 mAbs, huOKT3g1, aglycosyl CD3 and HuM291 have been used in limited trials in renal transplantation but have yet to have a formal clinical development. Humanized mAbs and receptor fusion proteins offer the potential of providing renal transplant recipients with a novel algorithm for immunosuppression that relies on chronic intermittent intravenous administration of safe, non-toxic agents replacing oral drug therapy maintenance. PMID- 12773966 TI - Daclizumab as induction therapy in kidney and simultaneous pancreas-kidney transplantation. AB - Acute rejection still remains a major problem in organ transplantation and is a significant risk factor for chronic rejection, and chronic rejection is one of the most important causes of late graft loss. Current new immunosuppressive drugs such as tacrolimus, rapamycin and mycophenolate mofetil have been developed to reduce acute rejection and to improve renal allograft survival. More recently, antihuman antibodies, either monoclonal or polyclonal, have been developed to use for induction therapy at the time of transplantation or to treat rejection. Daclizumab, a new engineered human immunoglobulin monoclonal antibody to the interleukin-2 receptor a-subunit was approved to prevent acute rejection after solid organ transplantation. Data from clinical trials show daclizumab to be well tolerated in solid organ transplantation. It does not increase the incidence of infection, including cytomegalovirus infection. PMID- 12773968 TI - Induction immunotherapy with IL-2Ra monoclonal antibody in kidney transplantation. AB - The development of new immunosuppressive agents is designed to reduce the incidence and severity of early acute post-transplant rejection. One potential target for more specific immunosuppressive therapy with monoclonal antibodies is the high affinity a chain of interleukin-2 receptors (IL-2Ra). Clinical investigation of murine IL-2Ra monoclonal antibodies (IL-2Ra mAb) in renal transplantation has indicated that a complete blockade of IL-2Ra during the critical first post-transplant months allows effective immunoprophylaxis, especially in the early post-transplant period. Efficacy of these agents, however, is hampered by their short disposition half-lives in humans and their immunogenicity in the form of neutralizing human antimouse antibodies. These inherent problems can be partially overcome by chi-meric, hyper-chimeric (humanized) products and multiple dose regimens. Both IL-2Ra mAbs: daclizumab (humanized) and basiliximab (chimeric) currently approved for clinical use have been found to reduce the frequency of acute rejections in renal transplant recipients without an apparent increase in short-term toxicities. In most transplant centers where these agents are utilized, they are being routinely administered as induction immunoprophylaxis in recommended multiple dose regimens to recipients of solid organ transplants. Others have restricted their use to certain high-risk patients such as those undergoing multi-organ transplantation, recipients with high panel-reactive antibodies, African-Americans, patients at risk for developing delayed graft function (DGF), and children. Recently some investigators have successfully administered these antibodies co-administered with newer immunosuppressive agents in limited dose protocols thus developing cost effective and simplified regimens. Therefore, in the absence of a favorable long-term efficacy, it is likely that these agents will be administered in limited dose protocols along with one of the modulators of IL-2, i.e. calcineurin inhibitors (CNI), to a selected group of patients in whom additional immunosuppression in the early post-transplantation period is desirable. PMID- 12773969 TI - Is cyclosporine withdrawal a therapeutic option in renal transplant recipients with chronic allograft dysfunction? AB - Chronic allograft nephropathy (CAN), the leading cause of graft loss, is due to both immunologic and non-immunologic factors. Cyclosporine (CsA) nephrotoxicity is supposed to significantly contribute to CAN. It has been suggested recently that new commercially available immunosuppressive drugs, such as Mycophenolate Mofetil (MMF) and Sirolimus (SRL), may have properties that may prevent and possibly partly reverse chronic rejection, the alloantigen-dependent immune process of CAN. Thus, our group and others have studied the efficiency of a 2 step therapeutic approach including CsA withdrawal and use of non-nephrotoxic immunosuppressive drugs with properties against chronic rejection, such as MMF or SRL. In this review, we will successively analyze how mechanisms of CsA-related nephrotoxicity, histopathologic findings in CAN, animal models and clinical studies justify this therapeutic approach. PMID- 12773970 TI - Mycophenolate mofetil in pediatric renal transplantation. AB - Since mycophenolate mofetil (MMF), an ester prodrug of the immunosuppressant mycophenolic acid (MPA), has been approved for maintenance immunosuppressive therapy also in children after renal transplantation it has become an important part of immunosuppressive protocols. By inhibiting inosine monophosphate dehydrogenase, the key enzyme in the de novo purine biosynthesis of proliferating T and B lymphocytes, MMF acts as a relatively specific inhibitor of human lymphocyte proliferation. MMF is more effective than azathioprine in combination with cyclosporin A (CsA) and corticosteroids and distinctly reduces the incidence of acute rejection episodes in the 1st year post-transplant in adults as well as in children. Beneficial effects on steroid-resistant rejection and chronic allograft dysfunction have been shown. In general, MMF is well tolerated. Major adverse events in pediatric renal transplant recipients include leukopenia, infections and gastrointestinal problems. Pharmacokinetic monitoring of MPA can help to optimise MMF therapy after renal transplantation, as associations between the risk of acute rejection episodes and MPA-AUC values and MPA predose levels have been demonstrated. The incidence of MMF-related side effects such as leukopenia and/or infections, however, is associated with pharmacokinetic parameters of free MPA. Reference data of relevant pharmacokinetic parameters are available. The possible steroid-sparing potential of MMF is an important issue in pediatric renal transplantation. Preliminary data demonstrate improved longitudinal growth, less cushingoid habitus and lower blood pressure after steroid-withdrawal in pediatric renal transplant recipients under MMF and CsA therapy. PMID- 12773971 TI - Hypertension after renal transplantation. AB - With current immunosuppression elevated blood pressure is found in almost 90% of renal graft recipients. Major causes of this finding are impairment of renal function, secondary to chronic allograft nephropathy or (less frequently) recurrence of primary renal disease, the use of calcineurin inhibitors as immunosuppresants, uncontrolled renin secretion by the shrunken kidneys of the recipient, stenos- ing lesions of the transplant artery (or the upstream arteries of the recipient), polycytemia and (genetic predisposition to) hypertension of the graft donor. Even minor degrees of blood pressure elevation have a significant impact on survival of the recipient and on graft survival, presumably by amplifying vascular injury to the graft. In this respect, elevation of systolic blood pressure and an abnormal circadian blood pressure profile are of particular relevance. In contrast to previous opinion, ACE inhibitors are indicated in the treatment, but, given the causal role of sodium retention in graft vasoconstriction, diuretics and calcium channel blockers remain main stays of antihypertensive treatment in the renal allograft recipient. PMID- 12773972 TI - The "new kid on the block": Intraductal papillary mucinous neoplasms of the pancreas: current concepts and controversies. PMID- 12773973 TI - Laparoscopic extraperitoneal inguinal hernia repair versus open mesh repair: a prospective randomized controlled trial. AB - BACKGROUND: This study was designed to compare an open tension-free technique (Lichtenstein repair) with a laparoscopic totally extraperitoneal hernia repair (TEP). METHODS: One hundred sixty-eight men aged 30 to 65 years with primary or recurrent inguinal hernia were randomized to TEP or open mesh technique in the manner of Lichtenstein. Follow-up was after 1 and 6 weeks, and 1 year. RESULTS: Eighty-one patients were randomized to TEP, and 87 to open repair. For 1 patient in each group, the operation was converted to a different type of repair. No difference was seen in overall complications between the 2 groups. However, 1 patient in the TEP group underwent operation for small bowel obstruction after surgery. A higher frequency of postoperative hematomas was seen in the open group (P <.05). Patients in the TEP group consumed less analgesic after surgery (P <.001), returned to work earlier (P <.01), and had a shorter time to full recovery (P <.01). Two recurrences occurred in the TEP group 1 year after surgery. CONCLUSION: The TEP technique was associated with less postoperative pain, a shorter time to full recovery, and an earlier return to work compared with the open tension-free repair. No difference was seen in overall complications. However, 2 recurrences did occur after 1 year in the TEP group. PMID- 12773974 TI - Presacral tumors: a practical classification and treatment of a unique and heterogeneous group of diseases. AB - BACKGROUND: Presacral tumors are a rare and diverse group of diseases that originate from the different tissues that comprise the potential presacral space. Because of their relative rarity, confusion exists regarding their clinical presentation, natural history, and treatment. The aim of this study is to describe a single institution's experience with the management of presacral tumors and to suggest a practical method of classification. METHODS: Records of all patients who underwent operation for presacral tumors from the years 1991 to 2001 were reviewed. Clinical, pathologic, treatment, and outcome variables were evaluated. RESULTS: Forty-two patients were included in the study and were divided into 4 groups according to lesion pathology: benign congenital (n = 12), malignant congenital (n = 9), benign acquired (n = 9), and malignant acquired (n = 12). Symptoms were nonspecific, and 26% of the cases were completely asymptomatic. Diagnosis was made with rectal examination and confirmed with pelvic computerized tomographic scan. Surgical approach varied among the different groups, with the posterior approach used mainly for congenital tumors and the anterior approach for acquired. Complete surgical resection of the tumor was obtained in all cases of benign tumors and in 76% of malignant tumors. No postoperative mortality was seen, and complications occurred in 36% (15/42); most were reversible. None of the patients with benign tumors had recurrences, and all are alive at this time. The survival rate of patients with malignant tumors was significantly improved when complete resection was possible. CONCLUSION: Classification of presacral tumors into congenital versus acquired and benign versus malignant is simple and efficient. Treatment is complete surgical resection, which can be performed safely with low morbidity and no mortality. PMID- 12773975 TI - Possible indication for surgical treatment of squamous cell carcinomas of the esophagus that involve the stomach. AB - BACKGROUND: The caudal spread of esophageal squamous cell carcinoma (ESCC) frequently involves the stomach. An extended surgical treatment may provide a tumor-free margin; however, its clinical benefit has not been elucidated. METHODS: Sixty-three of 582 patients with ESCC (11%) had massive gastric involvement and underwent esophagectomy with combined resection of the stomach and other organs. The mode of gastric involvement was classified as direct invasion from primary tumor (PT invasion) or invasion from metastatic lymph nodes (LN invasion). RESULTS: In addition to the removal of either the proximal (83%) or the whole (17%) stomach, 46 patients (73%) underwent the combined resection of adjacent organs, including the diaphragm, pancreas, liver, lung, and pericardium. This surgical treatment resulted in a high rate (83%) of curative resection and a low rate (8%) of operative mortality. Postoperative survival rates were 53%, 33%, and 25% at 1, 2, and 5 years, respectively. The first tumor recurrence was frequently in the abdominal paraaortic lymph nodes (41%) and the liver (28%), followed by the mediastinal lymph nodes, local recurrence, the lung, and other organs. The mode of gastric involvement strongly affected clinical outcome, with a 5-year survival rate of 36% for those with PT invasion but of only 7% with LN invasion (P <.0086). No significant difference was seen in the number and location of metastatic lymph nodes between the 2 groups; however, the size of the largest metastatic lymph node was significantly smaller with PT invasion than with LN invasion (12 mm vs 37 mm in diameter; P <.0001). CONCLUSION: Surgical treatment of ESCC involving the stomach was considered safe and successful. A favorable prognosis can be expected for gastric invasion from the primary tumor but not from metastatic lymph nodes. PMID- 12773976 TI - Prediction of survival with squamous cell carcinoma antigen in patients with resectable esophageal squamous cell carcinoma. AB - BACKGROUND: Increased preoperative serum squamous cell carcinoma antigen (SCC-Ag) concentrations have been found to be associated with advanced stage and poor prognosis in lung and cervical cancers. Because little was known about the significance of SCC-Ag concentration in patients with esophageal cancer, the aim of this study was to analyze the clinicopathologic significance of SCC-Ag in patients with esophageal SCC. PATIENTS AND METHODS. Preoperative SCC-Ag concentration was measured with enzyme-linked immunosorbent assay in 309 patients with primary esophageal SCC. All patients underwent curative radical surgery without any preoperative therapy. In 215 of 309 patients, carcinoembryonic antigen (CEA) was also measured to compare clinical significance of CEA with that of SCC-Ag. The prognostic significance for survival of SCC-Ag concentrations was studied with multivariate analysis with Cox proportional hazards model. RESULTS: The SCC-Ag concentration and the positivity rate of SCC-Ag were significantly elevated in patients associated with tumor progression. Statistically significant differences in SCC-Ag concentrations and SCC-Ag positivity rates were observed depending on tumor size, tumor depth, lymph node status, and distant metastasis. Although CEA was not a prognostic factor (P =.21), a high SCC-Ag concentration was a significant prognostic factor (P <.01). Multivariate analyses indicated that T factor had the best predictive power, but SCC-Ag concentration contained additional, independent prognostic information. CONCLUSION: Our findings suggest that preoperative serum SCC-Ag concentrations might provide a predictive information for tumor progression and survival in patients with esophageal SCC. PMID- 12773977 TI - Evaluation of preoperative portal embolization for safe hepatectomy, with special reference to assessment of nonembolized lobe function with 99mTc-GSA SPECT scintigraphy. AB - BACKGROUND: Preoperative portal embolization (PE) is used to stimulate liver hypertrophy in the nonembolized lobe. We studied liver volume and function with computed tomography and technetium-99m-galactosyl human serum albumin ((99m)Tc GSA) scintigraphy before PE and at 1 or 2 weeks after PE. METHODS: Right PE was performed in 30 patients. Morphologic and functional hypertrophy in the left lobe after PE was determined and related to the presence or absence of cholestasis, biliary drainage of the embolized lobe, and postoperative liver failure. RESULTS: The volume of the left lobe and (99m)Tc-GSA uptake increased rapidly for the first week after PE, but no significant increase was seen during the second week. Morphologic hypertrophy was less pronounced in patients with jaundice (P =.03). When PE was performed at a total bilirubin level above 2 mg/dL, the interval between PE and surgery was prolonged because of cholangitis and liver abscess formation. The net morphologic hypertrophy ratio was significantly higher in livers that had undergone left lobe drainage only (9.1% +/- 0.9%) compared with those in which there was drainage of the embolized lobes (5.7% +/- 0.9%; P =.03). The volume and (99m)Tc-GSA uptake of the left lobe in the second week after PE was significantly smaller in patients with postoperative liver failure (33.7% +/- 2.4% and 18.0% +/- 2.1%, respectively) than in patients without liver failure (46.2% +/- 1.4% and 38.4% +/- 2.3%; P =.003 and P =.01, respectively). CONCLUSION: In the nonembolized lobe, the functional increase in (99m)Tc-GSA uptake is more pronounced than suggested by the degree of morphologic hypertrophy. Whenever possible, biliary drainage should not be performed in the lobe undergoing hepatectomy. (99m)Tc-GSA SPECT scintigraphy is useful for the evaluation of postoperative liver failure. PMID- 12773978 TI - Surgical resection of liver metastases of gastric cancer: an analysis of a 17 year experience with 22 patients. AB - BACKGROUND: The justification for surgical resection of liver metastases from gastric cancer remains controversial. METHODS: Twenty-two patients who underwent 26 hepatectomies for liver metastases of gastric cancer between 1985 and 2001 were analyzed. Fifteen clinicopathologic factors were evaluated with univariate and multivariate analyses for survival after hepatic resection. RESULTS: The overall 1-year, 3-year, and 5-year survival rates after hepatectomy for gastric metastases were 73%, 38%, and 38%, respectively. Five patients survived for more than 3 years without recurrence, 3 of whom had synchronous metastases resected at the time of gastrectomy. The best results after surgical resection for liver metastases of gastric cancer were obtained with solitary metastases less than 5 cm in size. The number of liver metastases (solitary or multiple) was the only significant prognostic factor according to both univariate and multivariate analyses. CONCLUSION: Surgical resection for liver metastases of gastric cancer may be beneficial for patients with a solitary metastasis, whether it is synchronous or metachronous. PMID- 12773979 TI - Biliary bile acid concentration is a simple and reliable indicator for liver function after hepatobiliary resection for biliary cancer. AB - BACKGROUND: The functional recovery of the remnant liver after an extended hepatectomy is critical for the outcome of the patient. The aim of this prospective study was to examine whether biliary bile acids could be an indicator for postoperative liver function. METHODS: Externally drained bile samples were obtained from 51 patients with biliary or periampullary carcinomas before and after surgery. Patients were categorized into 3 groups: group A, 29 hepatectomized patients without liver failure; group B, 7 hepatectomized patients with liver failure (maximum serum bilirubin level, >10 mg/dL); and group C, 15 patients who underwent biliopancreatic resection without hepatectomy, with a good postoperative course. Bile samples were withdrawn 1 day before surgery and on postoperative days 1, 2, 3, 4, 6, and 7. Total bile acids were measured with a 3 alpha-hydroxysteroid dehydrogenase method. RESULTS: Before surgery, the concentration of bile acids was higher in groups A and C than in group B, and correlated significantly with the indocyamine green disappearance rate (KICG) values (R(2) = 0.557; P <.0001). After surgery, bile acid concentrations decreased in all 3 groups until postoperative day 2, which was followed by a gradual increase. The concentration recovered to the preoperative level in groups A and C but remained low in group B. Biliary bile acid concentrations on day 2 correlated significantly with remnant liver KICG values (R(2) = 0.257; P =.0019). Among several parameters studied, including KICG, remnant liver KICG, biliary bile acids, and biliary bilirubin, biliary bile acid concentration had the most predictive power for occurrence of postoperative liver failure. CONCLUSION: Biliary bile acid concentration could be a simple, real-time, reliable indicator of preoperative and postoperative liver function. PMID- 12773980 TI - Radical antegrade modular pancreatosplenectomy. AB - INTRODUCTION: Retrograde distal pancreatectomy with splenectomy is the standard procedure for cancers of the body and tail of the pancreas. However, this procedure has limitations in terms of the posterior extent of resection and the ability to achieve a complete N1 node resection. METHODS: A new antegrade procedure has been developed that provides improved visibility, removes N1 nodes, and permits adjustment of the depth of the posterior extent of resection coupled with early rather than late control of the vasculature. RESULTS: Ten patients, 6 with adenocarcinomas of the body of the pancreas, have undergone the procedure since 1999. Nine of 10 patients had negative resection margins, and the median node count in patients who did not receive neoadjuvant radiation was 9 nodes. Three patients had complications develop; no postoperative deaths occurred. CONCLUSION: Early results with the procedure are encouraging. PMID- 12773981 TI - Inhibition of inducible nitric oxide synthase results in reductions in wound vascular endothelial growth factor expression, granulation tissue formation, and local perfusion. AB - BACKGROUND: Wound repair results from a series of highly orchestrated cellular and biochemical events, including increased synthesis of the bioregulatory molecule nitric oxide (NO). The goal of this work was to test the functional role of NO in promotion of vascular endothelial growth factor (VEGF) production and the vigorous granulation tissue formation characteristic of this wound model. METHODS: A ventral hernia, surgically created in the abdominal walls of 12 swine, was repaired with silicone sheeting and skin closure. An osmotic infusion pump, inserted in a remote subcutaneous pocket, delivered saline solution (n = 6) or the selective inducible NO synthase inhibitor N(6) (iminoethyl)-L-lysine (L-NIL; n = 6) into the wound environment. Granulation tissue thickness was determined with ultrasonography, and local wound perfusion was measured with laser Doppler analysis for 2 weeks. Fluid was aspirated serially from the wound compartment for measurement of nitrite/nitrate, VEGF, and transforming growth factor beta(1)concentrations. On day 14, the animals were killed and the abdominal wall was harvested for immunohistochemical and molecular analysis. RESULTS: In animals that received saline solution, a nearly linear 4-fold increase in granulation tissue thickness was measured during the 14-day interval. In contrast, in animals that received L-NIL, day 14 granulation tissue thickness was essentially unchanged from the day 2 values of saline solution-treated animals. Moreover, in the L-NIL animals, ultrasonography was unable to resolve the angiogenic zone typical of controls, and correspondingly, wound vessel count and vascular surface area estimates derived from image analysis of histologic sections were 2-fold to 3-fold lower in the L-NIL animals compared with controls. Reductions in basal (2 fold) and heat-provoked (2.5-fold) wound perfusion were noted in L-NIL animals. Wound fluid nitrite/nitrate and VEGF levels were strikingly (4-fold and 5-fold, respectively) reduced in L-NIL animals on days 9 to 14. Immunochemistry results showed reduced VEGF protein content in granulation tissue and keratinocytes within the hyperproliferative epithelium at wound edge. Finally, transforming growth factor-beta(1)levels were unaffected by L-NIL treatment. CONCLUSION: VEGF production in granulation tissue is dependent on the presence of functionally active inducible NO synthase and hence, the production of NO. NO and VEGF are therefore defined as key regulators of granulation tissue formation. PMID- 12773982 TI - Relative hyperoxia augments lipopolysaccharide-stimulated cytokine secretion by murine macrophages. AB - BACKGROUND: Increased systemic levels of inflammatory mediators are seen after open abdominal operations. Macrophages that are exposed to lipopolysaccharide secrete cytokines. Peritoneal macrophages normally reside in a pO(2) of 40 mm Hg. We hypothesize that exposure of lipopolysaccharide-stimulated macrophages to "non physiologic" pO(2) augments cytokine secretion. METHOD: Murine macrophages were preconditioned to a pO(2) of 40 mm Hg for 24 hours. The medium then was discarded and exchanged for a medium containing a pO(2) of 40, 150, or 440 mm Hg. Macrophages were incubated in the desired pO(2) for 6 and 24 hours while stimulated with lipopolysaccharide (0 to 100 ng/mL). The effect of pO(2) was compared. Supernatant tumor necrosis factor (TNF) and interleukin-6 were measured with enzyme-linked immunosorbent assay. Statistics were performed with analysis of variance. RESULTS: We found dose-dependent lipopolysaccharide-stimulated TNF and interleukin-6 production with macrophages incubated at physiologic pO(2). Higher pO(2) did not stimulate TNF and interleukin-6 in the absence of lipopolysaccharide. However, a pO(2) of 150 and 440 mm Hg significantly (P <.05) increased lipopolysaccharide-stimulated TNF and interleukin-6 production versus 45 mm Hg. CONCLUSION: Our data suggest synergy between increased pO(2) and lipopolysaccharide for macrophage TNF and interleukin-6 production. Similar pO(2) elevations may occur with an open peritoneum or high supplemental O(2). Cytokines from peritoneal macrophages may contribute to the increased systemic inflammation after open operations. PMID- 12773983 TI - Increased surgical stress promotes tumor metastasis. AB - BACKGROUND: Although it is well-known that excessive surgical stress augments the growth of residual cancer and metastasis, whether surgical stress is increased according to the degree of surgical manipulation and can consequently lead to the enhancement of cancer metastasis has not been thoroughly examined. Moreover, the molecules associated with response for stress-enhanced metastasis have not been well-analyzed. The aim of this study was to examine whether cancer metastasis is enhanced with an increase of surgical stress with an experimental lung metastasis model and to analyze the related molecules responsible for stress-enhanced metastasis. METHODS: Colon 26-L5 carcinoma cells (1.5 x 10(4)/mouse) were injected intravenously into 6-week-old female BALB/c mice (Japan SLC, Hamamatsu, Japan). Two hours later, the mice were divided into 5 groups: untreated controls (the C group); mice given anesthesia only (the A group); mice given anesthesia and laparotomy (the AL group); mice given anesthesia, laparotomy, and appendectomy (the ALAp group); and mice given anesthesia, laparotomy, appendectomy, and left hepatic lobectomy (the ALApH group). The anesthesia procedures were the same in all groups (intraperitoneal administration of 0.8 mg/mouse sodium pentobarbital). In the AL, ALAp, and ALApH groups, a 3-cm long laparotomy was performed, and the time of the whole operation was just 5 minutes. All mice were killed 14 days after the procedures, and the number of lung metastases on the lung surface was counted manually. At the same time, BALB/c mice without tumor burden were given the same 5 kinds of surgical stress, and the messenger RNA expression of various metastasis-related molecules in the lung was measured with reverse transcriptase-polymerase chain reaction at 6, 24, and 48 hours after surgical stress. We also examined the effect of ONO-4817 (an inhibitor of matrix metalloproteinases ([MPs]) on lung metastasis in the mice with the 5 kinds of surgical stress. RESULTS: The numbers of lung metastases on the lung surface and the messenger RNA expression of MMP-9, membrane type IBMMP, and urokinase-type plasminogen activator at 24 hours after surgery were enhanced in proportion to the degree of surgical stress. Moreover, ONO-4817 significantly inhibited lung metastasis. CONCLUSION: These results strongly suggest that increased surgical stress augments cancer metastasis via surgical stress-induced expression of proteinases in the target organ of metastasis. PMID- 12773984 TI - Pyrrolidine dithiocarbamate provides protection against hypothermic preservation and transplantation injury in the rat liver: the role of heme oxygenase-1. AB - BACKGROUND: Pyrrolidine dithiocarbamate (PDTC) represents a class of antioxidants and is a potent inducer of the heme oxygenase-1 (HO-1) gene and an inhibitor of nuclear factor-kappa B (NF-kappa B). We examined the impact of PDTC preconditioning against cold ischemia and reperfusion injury in the rat liver. METHODS: Lewis rats were treated subcutaneously with saline or PDTC solution 24 hours before harvesting. Some animals pretreated with PDTC were also given zinc protoporphyrin IX intravenously immediately after reperfusion. HO-1 expression and enzyme activity in liver tissues were analyzed at different time points after each treatment. After transplantation of 24-hour preserved livers, serum levels of transaminases and gene expression of tumor necrosis factor-alpha, interleukin 1 beta, and NF-kappa B were measured. Animal survival and cellular viability were monitored. RESULTS: HO-1 gene expression and protein synthesis were enhanced in PDTC-treated livers, leading to increased enzyme activity (P <.05). The PDTC treatment group showed lower transaminase levels (P <.05), lower cytokine and NF kappa B messenger RNA expression (P <.05), and fewer nonviable cells (P <.05) than did the control group, whereas these PDTC effects were abolished with zinc protoporphyrin injection after reperfusion (P <.05). The best animal survival rate was observed in the PDTC group (P <.05). CONCLUSION: PDTC preconditioning reduces inflammatory responses during reperfusion. PDTC appears to exert this protective effect by induction of an antioxidative stress protein and inhibition of proinflammatory cytokines. PMID- 12773986 TI - Left hepatic trisegmentectomy with right hepatic vein resection after right hepatic vein embolization. PMID- 12773985 TI - Intestinal transformation results in transforming growth factor-beta-dependent alteration in tumor cell-cell matrix interactions. AB - BACKGROUND: An alteration in the expression of and response to transforming growth factor-beta 1 (TGF-beta 1) appears to be an important event during colorectal carcinogenesis. However, the precise role of TGF-beta 1 in colorectal carcinogenesis is not clear. We have previously described in detail the changes in cell proliferation and differentiation caused by chronic exposure to TGF-beta 1. In this study we sought to better characterize the changes in tumor cell-cell matrix interactions seen during TGF-beta 1-mediated intestinal transformation. METHODS: Rat intestinal epithelial cells (RIE) and RIE cells transformed by chronic exposure to TGF-beta 1 (RIE-Tr) were treated with TGF-beta 1 and production of components of the plasmin/plasminogen system measured by ELISA and Western blotting. TGF-beta 1 effects on invasion and adhesion were determined in vitro. The role of urokinase on TGF-beta 1-mediated invasion and adhesion were determined using immunoneutralization. The role of COX-2 was determined using a specific COS-2 inhibitor. RESULTS: TGF-beta 1 had no effect on RIE-1 adhesion to collagen types I and IV, fibronectin, and laminin, or invasion through collagen types I and IV. However, 5 ng/mL TGF-beta 1 significantly increased the invasiveness and decreased the adhesiveness of RIE-Tr. This effect of TGF-beta 1 on RIE-Tr was associated with a significant increase in plasmin activity secondary to increased expression of uPA. TGF-beta 1 had no effect on either uPA receptor or PAI-1 in this system. Antibodies to uPA completely blocked the TGF beta 1-mediated invasiveness of the RIE-Tr cells and returned their adhesiveness to basement membrane proteins to baseline. Addition of the selective Cox-2 inhibitor SC-58125 resulted in a dose-dependent decrease in TGF-beta 1-mediated invasion and uPA expression. CONCLUSION: This study provides additional evidence for TGF-beta 1 as a tumor promoter during intestinal carcinogenesis and a possible new mechanism for Cox-2-related colon carcinogenesis. PMID- 12773987 TI - The function of the spleen in adults after ligation of the splenic artery of the traumatized spleen in childhood. AB - BACKGROUND: Ligation of the splenic artery (LSA) has been successfully used as a spleen-saving procedure in rare cases of splenic trauma in children in which management with splenorrhaphy or partial splenectomy alone was not possible. There are no data regarding the long-term effects of the procedure on the functional status of the spleen. The purpose of this study is to present and discuss our clinical and laboratory findings in adults who underwent LSA in childhood. METHODS: Our first 2 patients in whom LSA was done at ages 4 and 2 years in 1977 underwent the following examinations in the year 2000: 1, imaging of the spleen; 2, immunologic studies; and 3, peripheral blood tests. Their ages at reexamination were 27 and 25 years, respectively. RESULTS: Results were as follows: triplex ultrasound revealed normal size and echomorphology; Doppler techniques revealed normal vasculature; 99mTc-Tin colloid scanning revealed normal uptake. Immunoglobulins (IgG1 to IgG4, IgA, IgM, IgE), complement fraction (C3, C4), antibodies response to vaccinations, and peripheral blood tests all had normal results. No Howell-Jolly bodies were found. CONCLUSION: Laboratory investigations in adults with LSA during childhood disclosed undisturbed function of the spleen. LSA can be used as an adjunct to splenorrhaphy in children with rare splenic injuries involving major hilar vessels. PMID- 12773988 TI - Zigzag seams for the prevention of scar bands after sheet split-thickness skin grafting. PMID- 12773989 TI - Spurious precision. PMID- 12773990 TI - Colonel Pete. PMID- 12773991 TI - Intractable hematochezia: an unusual presentation of intestinal tuberculosis. PMID- 12773992 TI - Hepatic angiomyolipoma. PMID- 12773993 TI - Cystic nephroma: a benign renal tumor of children and adults. PMID- 12773994 TI - One death after radiofrequency thermal ablation for hepatocellular carcinoma in a cirrhotic patient. PMID- 12773995 TI - Choice of statistical test, and choice of research question. PMID- 12773996 TI - Aberrant inferior thyroid artery: two cases. PMID- 12773997 TI - Pitfalls on frozen section of a hepatic lesion in the management of a pancreatic tumor. PMID- 12773998 TI - Hepatic copper metabolism: insights from genetic disease. PMID- 12773999 TI - Hepatic stem cells and developmental biology of the liver. PMID- 12774001 TI - Nonselective beta-blockers plus nitrates in portal hypertension: an open question. PMID- 12774000 TI - HNF4: a central regulator of hepatocyte differentiation and function. PMID- 12774002 TI - Viral kinetics in hepatitis C. PMID- 12774004 TI - A role of autoantibody-mediated platelet destruction in thrombocytopenia in patients with cirrhosis. AB - Thrombocytopenia is a common manifestation in patients with liver cirrhosis (LC), but its underlying mechanism remains controversial. This study examined the role of anti-platelet autoimmunity in cirrhotic thrombocytopenia by determining the autoantibody response to GPIIb-IIIa, a major platelet surface autoantigen recognized by anti-platelet antibodies in patients with idiopathic thrombocytopenic purpura (ITP). Circulating B cells producing anti-GPIIb-IIIa antibodies as well as platelet-associated and plasma anti-GPIIb-IIIa antibodies were examined in 72 patients with LC, 62 patients with ITP, and 52 healthy controls. In vitro anti-GPIIb-IIIa antibody production was induced in cultures of peripheral blood mononuclear cells (PBMCs) by stimulation with GPIIb-IIIa. The frequency of anti-GPIIb-IIIa antibody-producing B cells in patients with LC was significantly greater than in healthy controls (10.9 +/- 6.2 vs. 0.4 +/- 0.3/10(5) PBMCs; P <.0001) and was even higher than the frequency in patients with ITP (8.2 +/- 5.2; P =.007). Anti-GPIIb-IIIa antibodies in the patients with LC and ITP were mainly present on the surfaces of circulating platelets rather than in the plasma in an unbound form. Furthermore, PBMCs from patients with LC and ITP produced anti-GPIIb-IIIa antibodies on antigenic stimulation with GPIIb IIIa in vitro, and the specific antibodies produced had the capacity to bind normal platelet surfaces. In conclusion, the similar profile of the anti-GPIIb IIIa autoantibody response in patients with LC and ITP suggests that autoantibody mediated platelet destruction may contribute at least in part to cirrhotic thrombocytopenia. PMID- 12774003 TI - Propranolol plus placebo versus propranolol plus isosorbide-5-mononitrate in the prevention of a first variceal bleed: a double-blind RCT. AB - Nonselective beta-blockers are very effective in preventing first variceal bleeding in patients with cirrhosis. Treatment with isosorbide-5-mononitrate (IS MN) plus propranolol achieves a greater reduction in portal pressure than propranolol alone. The present multicenter, prospective, double-blind, randomized, controlled trial evaluated whether combined drug therapy could be more effective than propranolol alone in preventing variceal bleeding. A total of 349 consecutive cirrhotic patients with gastroesophageal varices were randomized to receive propranolol + placebo (n = 174) or propranolol + IS-MN (n = 175). There were no significant differences in the 1- and 2-year actuarial probability of variceal bleeding between the 2 groups (propranolol + placebo, 8.3% and 10.6%; propranolol + IS-MN, 5% and 12.5%). The only independent predictor of variceal bleeding was a variceal size greater than 5 mm. However, among patients with varices greater than 5 mm (n = 196), there were no significant differences in the incidence of variceal bleeding between the 2 groups. Survival was also similar. Adverse effects were significantly more frequent in the propranolol + IS-MN group due to a greater incidence of headache. There were no significant differences in the incidence of new-onset or worsening ascites or in impairment of renal function. In conclusion, propranolol effectively prevents variceal bleeding. Adding IS-MN does not further decrease the low residual risk of bleeding in patients receiving propranolol. However, the long-term use of this combination drug therapy is safe and may be an alternative in clinical conditions associated with a greater risk of bleeding. PMID- 12774005 TI - The kidney plays a major role in the hyperammonemia seen after simulated or actual GI bleeding in patients with cirrhosis. AB - Upper gastrointestinal (UGI) bleeding in cirrhosis is associated with enhanced ammoniagenesis, the site of which is thought to be the colon. The aims of this study were to evaluate interorgan metabolism of ammonia following an UGI bleed in patients with cirrhosis. Study 1: UGI bleed was simulated in 8 patients with cirrhosis and a transjugular intrahepatic portasystemic stent-shunt (TIPSS) by intragastric infusion of an amino acid solution that mimics the hemoglobin molecule. We sampled blood from the femoral artery and a femoral, renal, portal, and hepatic vein for 4 hours during the simulated bleed and measured plasma flows across these organs. Study 2: In 9 cirrhotic patients with an acute UGI bleed that underwent TIPSS insertion, blood was sampled from an artery and a hepatic, renal, and portal vein, and plasma flows were measured. Study 1: During the simulated bleed, arterial concentrations of ammonia increased significantly (P =.002). There was no change in ammonia production from the portal drained viscera, but renal ammonia production increased 6-fold (P =.008). In contrast to an unchanged ammonia removal by the liver, a significant increase in muscle ammonia removal was observed. Study 2: In patients with an acute UGI bleed, ammonia was only produced by the kidneys (572 [184] nmol/kg bw/min) and not by the splanchnic area (-121 [87] nmol/kg bw/min). In conclusion, enhanced renal ammonia release has an important role in the hyperammonemia that follows an UGI bleed in patients with cirrhosis. During this hyperammonemic state, muscle is the major site of ammonia removal. PMID- 12774006 TI - Clinical and histologic spectrum of nonalcoholic fatty liver disease associated with normal ALT values. AB - A retrospective study was performed to (1) characterize the clinical and histologic features of those with nonalcoholic fatty liver disease (NAFLD) and normal alanine aminotransferase (ALT) values, (2) compare the spectrum of NAFLD associated with normal versus elevated ALT levels, and (3) determine whether there were differences in the clinical or histologic spectrum of NAFLD between those with a low normal versus high normal ALT value. A total of 51 subjects with NAFLD and normal ALT were identified and compared with 50 consecutive subjects with NAFLD and elevated ALT. The major indications for liver biopsy in those with normal ALT were unexplained hepatomegaly (n = 21) and evaluation as a potential donor for living donor liver transplantation (n = 16). The 2 groups were comparable with respect to age, gender distribution, and ethnicity. Approximately 80% of cases in both groups had at least 1 feature of the metabolic syndrome, the major risk factor for NAFLD. The 2 groups were also comparable with respect to the grade of the individual histologic parameters of NAFLD. A total of 12 subjects with normal ALT levels had bridging fibrosis, whereas 6 had cirrhosis. Diabetes was the only factor independently associated with an increased risk of advanced fibrosis (bridging fibrosis or cirrhosis) by multivariate analysis (relative risk: 2.3, P <.01). The mean steatosis (1.6 vs. 2.16, P <.04) and perisinusoidal fibrosis scores (0.35 vs. 0.9, P <.049) were lower in those with low normal (<30 IU/L) ALT versus high normal ALT. However, the prevalence of advanced fibrosis was similar (5 of 15 vs. 13 of 36, respectively). In conclusion, (1) the entire histologic spectrum of NAFLD can be seen in individuals with normal ALT values, (2) the histologic spectrum in these individuals is not significantly different from those with elevated ALT levels, and (3) a low normal ALT value does not guarantee freedom from underlying steatohepatitis with advanced fibrosis. PMID- 12774007 TI - All liver recipients benefit from the protocol 10-year liver biopsies. AB - The value of late protocol biopsies after liver transplantation remains to be evaluated to highlight the therapeutic policies. The study population was composed of patients who survived with the initial graft and with an available 10 year protocol biopsy (n = 143). The long-term histologic outcome of the graft, particularly the rate of ductopenia in cases with chronic rejection (CR), and Metavir scoring of fibrosis in cases with viral chronic hepatitis (VCH), were assessed. Fibrosis progression (FP) rates were compared over 3 periods (0-5, 5 10, and 0-10 years). At 10 years, histologic abnormalities present in 80% of the patients were not identifiable from liver function tests (LFTs), which were strictly normal in 52% of the patients. Histologic CR occurred in 24% at 10 years, with a mean rate of ductopenia higher at 10 years than at 5 years (49% vs. 34%, P <.001). In cases of VCH, fibrosis worsened, with a median FP rate of 0.20 fibrosis units/year. During the first 5 years, FP was as follows; hepatitis B virus infection was greater than recurrent hepatitis C virus (HCV) infection, which was greater than acquired HCV infection (P =.029). In patients with HCV, FP was higher during the second 5-year period than during the first one (P =.042). In conclusion, given the high prevalence of histologic abnormalities and the lack of sensitivity and specificity of LFTs, late protocol biopsies clearly are justified to adjust treatments, not only in HCV-infected patients in whom FP was fast and not linear, but also in the whole population of recipients. PMID- 12774008 TI - Portal cavernoma-associated cholangiopathy: a clinical and MR cholangiography coupled with MR portography imaging study. AB - Although biliary symptoms appear to be uncommon in patients with portal cavernoma, almost all of them have endoscopic retrograde cholangiographic abnormalities. The mechanisms underlying the biliary changes are explained poorly. This study in patients with portal cavernoma had 3 aims: (1) to assess the manifestations related to biliary involvement; (2) to evaluate with magnetic resonance (MR) imaging the aspect and frequency of cholangiographic changes; and (3) to clarify the mechanisms underlying biliary involvement. From December 1999 to July 2001, 25 consecutive adults with portal cavernoma without cancer or cirrhosis were studied with MR cholangiography coupled with MR portography. Seven patients presented with clinical manifestations of biliary disease. MR cholangiography findings were stenosis in 21 patients, with upstream dilatation in 16 and displacement in 13. MR cholangiography coupled with MR portography showed in all cases that the biliary abnormalities were secondary to a mass effect directly related to pressure by the cavernoma. In conclusion, in patients with portal cavernoma, clinical manifestations of biliary disease are most frequent; the cholangiographic abnormalities are very common and usually are caused by a mass effect by pressure from the veins composing the cavernoma. MR cholangiography coupled with MR portography is an effective noninvasive examination for simultaneous visualization of bile ducts and their relationship to the cavernoma. PMID- 12774009 TI - Serum HBV DNA as a marker of efficacy during therapy for chronic HBV infection: analysis and review of the literature. AB - Currently, compounds under evaluation for treatment of chronic hepatitis B virus (HBV) infection are evaluated with liver histology as the primary end point for efficacy. However, because of practical limitations in serial liver biopsies, there is a need for alternate markers to assess efficacy over shorter periods of time. Considering the direct correlation between viral replication and disease progression during human immunodeficiency virus and hepatitis C virus infection, we explored whether such a correlation exists for HBV infection. We reviewed the literature and conducted an analysis to investigate the relationship between absolute or treatment-induced changes in HBV DNA levels and other accepted markers of disease activity. A total of 26 prospective studies met our selection criteria, including 33 evaluable treatment arms. The study treatments consisted of nucleosides and/or interferon regimens and control arms. We found statistically significant and consistent correlations between viral load level or change and histologic grading and biochemical and serologic response. Our analysis suggests that a treatment-induced reduction in HBV DNA level can be used for assessing efficacy of treatment regimens. Further, we observed that quantitative HBV DNA has a broader dynamic range than histology, allowing demonstration of differences between 2 active treatments of unequal potency. The analysis showed stronger results in studies using nucleoside regimens and in hepatitis B e antigen (HBeAg)-positive patients. In conclusion, the goal of anti HBV therapy should be profound and durable viral suppression, as defined by very sensitive assays. Additional prospective studies are needed to precisely determine the desirable level of viremia to attain. PMID- 12774010 TI - Steroid-free chemotherapy decreases risk of hepatitis B virus (HBV) reactivation in HBV-carriers with lymphoma. AB - Reactivation of hepatitis is one of the most serious complications of chemotherapy in lymphoma patients who are carriers of the hepatitis B virus (HBV). Glucocorticoids are linked to increased risk of HBV reactivation. This study seeks to clarify whether removal of glucocorticoids from chemotherapy regimens may decrease the risk of HBV reactivation. Eligible patients were seropositive for hepatitis B surface antigen (HBsAg) and had histologically proven non-Hodgkin's lymphomas for which intensive chemotherapy was indicated. Patients were randomized to receive either ACE (epirubicin, cyclophosphamide, and etoposide) or PACE (prednisolone + ACE). A total of 50 patients were enrolled, 25 each for the ACE and PACE arms. The cumulative incidence of HBV reactivation at 9 months after starting chemotherapy was 38% and 73% for ACE and PACE arm, respectively (P =.03). The degree of clinical hepatitis was significantly more severe in the PACE arm: 11 patients (44%) in the PACE and 3 patients (13%) in the ACE arm had ALT elevation more than 10-fold of normal (P =.025), and 7 patients (28%) in the PACE and 1 patient (4%) in the ACE arm had icteric hepatitis (P =.049). Complete remission of tumors occurred in 11 (46%) patients in the PACE and 8 (35%) patients in the ACE arm (P =.556). The estimated overall survival rate at 46 months was 68% in the PACE arm and 36% in the ACE arm, respectively (P =.18). In conclusion, steroid-free chemotherapy decreases the incidence and severity of HBV reactivation in HBsAg-positive lymphoma patients. However, further research is needed to evaluate whether steroid-free chemotherapy may confer a less satisfactory control of lymphoma. PMID- 12774011 TI - Active immunization against de novo hepatitis B virus infection in pediatric patients after liver transplantation. AB - The shortage of donor organs occasionally mandates the use of hepatic allografts from anti-HBc (+) donors. HBIG and/or lamivudine are recommended for the prevention of de novo HBV infection in naive patients, but there are attendant problems, such as mutant strain emergence and high cost. Active immunization presents a better alternative than the use of HBIG or lamivudine, if it can be proven to be effective. Accordingly, we investigated the outcome of HBV vaccination in pediatric hepatic transplant recipients. Between July 1999 and October 2001, 19 pediatric recipients were administered HBV vaccinations after liver transplantation at Seoul National University Hospital. Nine patients received a graft from anti-HBc (+) donors and 10 from anti-HBc (-) donors. When steroid was withdrawn, recombinant HBV vaccine was administered. The median follow-up period after vaccination was 10.0 +/- 5.2 months. Seventeen of the 19 patients showed a positive response to vaccination. In 9 patients who received grafts from anti-HBc (+) donors, 2 patients showed no response, 4 patients low response (peak HBsAb titer <1,000 IU/L), and 3 patients high response (peak HBsAb titer >/=1,000 IU/L). De novo HBV infection developed in 1 of 2 patients who showed no response to vaccination. In 10 patients who received grafts from anti HBc (-) donors, 5 showed a low response and 5 a high response. In conclusion, HBV vaccination in pediatric patients after liver transplantation appeared to exhibit some effectiveness at protecting young children that received a graft from anti HBc (+) donors from de novo HBV infection. PMID- 12774012 TI - CD8+ T-cell interaction with HCV replicon cells: evidence for both cytokine- and cell-mediated antiviral activity. AB - The interaction between the host immune response and infected hepatocytes plays a central role in the pathogenesis of hepatitis C virus (HCV). The lack of a suitable animal or in vitro model has hindered our understanding of the host T cell/HCV interaction. Our aim was to develop an in vitro model to study the mechanisms of HCV-specific T-cell-mediated antiviral and cytolytic function. The HCV replicon was HLA typed and lymphocytes were obtained from an HLA class I matched subject. CD8(+) T cells were expanded with 2 HCV-specific/HLA-restricted peptides for NS3. Lymphocyte preparations were cocultured with HCV replicon (FCA1) and control (Huh7) cells labeled with (51)Cr. After a 48-hour incubation, the cells were harvested for RNA extraction. Standard blocking assays were performed in the presence of anti-interferon gamma (IFN-gamma), anti-tumor necrosis factor alpha (TNF-alpha), and anti-FasL. Cytolytic activity was measured by (51)Cr release. HCV replicon cells express homozygous HLA-A11 alleles and present HCV nonstructural proteins. HCV-specific expansion of CD8(+) cells led to a 10-fold decrease in HCV replication by Northern blot analysis and 21% specific lysis of FCA1 cells (compared with 2% of control Huh7 cells). Twenty percent of this antiviral activity was independent of T-cell binding, suggesting cytokine mediated antiviral activity. The CD8(+) antiviral effect was markedly reduced by blocking either IFN-gamma or FasL but was unaffected by blocking TNF-alpha. In conclusion, HCV-specific CD8(+) cells inhibit viral RNA replication by cytokine mediated and direct cytolytic effects. This T-cell/HCV subgenomic replicon system represents a model for the investigation of CD8 cell interaction with HCV infected hepatocytes. PMID- 12774013 TI - Viral dynamics and response differences in HCV-infected African American and white patients treated with IFN and ribavirin. AB - Studies have suggested that African American patients infected with hepatitis C virus (HCV) do not respond as well to treatment with interferon (IFN) as white patients. Here we analyzed the difference in the viral kinetic response between genotype 1 HCV-infected African American patients (n = 19) and white patients (n = 16). Patients were treated with 10 mIU IFN-alpha2b daily with or without ribavirin for 1 month followed by 3 mIU IFN-alpha2b 3 times a week with ribavirin. The kinetic parameters (epsilon, treatment effectiveness at inhibiting virion production; delta, loss rate of virus-producing cells; c, clearance rate of free virions; tau, delay until viral decline starts) were estimated from the viral load decay profiles using a previously described mathematical model. Differences in early kinetic parameters and viral negativity frequencies at weeks 4, 12, and 48 were compared. Ribavirin did not appear to enhance any of the viral kinetic parameters, although this may have been due to the high dose of IFN used. African American patients exhibited significantly (P =.005) lower drug effectiveness (88.6% vs. 98.2%) compared with white patients, accounting for a 0.8 log lower HCV RNA decrease in the first 24 hours of treatment. Significant differences (P =.006) were also noted for delta. There was no correlation between any of the viral kinetic parameters and either age, body mass index (BMI), or genotype 1 subtype. No patient achieved viral negativity at weeks 4, 12, or 48 without an epsilon greater than 90%. The mean viral decline and viral negativity rates were statistically different between the two races; however, when controlling for treatment effectiveness, these differences were no longer apparent. In conclusion, the failure of IFN response in African American patients infected with genotype 1 HCV is in part due to an impaired ability to inhibit viral production. PMID- 12774014 TI - Effect of ribavirin on hepatitis C viral kinetics in patients treated with pegylated interferon. AB - A dynamic equilibrium between viral production and clearance characterizes untreated chronic hepatitis C viral infection. After initiating antiviral treatment, a typical multiphasic decay of viremia can be observed and analyzed using mathematical models. To elucidate the antiviral mechanism of ribavirin when used in combination with (pegylated) interferon alfa, we investigated kinetic parameters in patients with chronic hepatitis C treated with either peginterferon alpha-2a with or without ribavirin and standard interferon alpha-2b plus ribavirin for 48 weeks. Serum HCV RNA was measured frequently before, during, and at the end-of-treatment and the follow-up period. By using an appropriate model for viral dynamics, kinetic parameters were derived from nonlinear, least square fitting of serum HCV RNA quantifications. The first phase of viral decay (day 1) and the second phase of viral decay (days 2 to 21) were similar for all treatment groups. After about 7 to 28 days, a third phase of viral decay was seen in several patients, and this phase of decay was significantly faster in patients treated with peginterferon alpha-2a plus ribavirin compared with those treated with peginterferon alpha-2a alone. The decay of this third phase was associated with the virologic end-of-treatment response and sustained virologic response. In conclusion, the third-phase decay of initial viral kinetics, which may represent a treatment-enhanced degradation of infected cells, was more pronounced in patients treated with peginterferon alpha-2a plus ribavirin. This finding suggests that combination treatment leads to a better restoration of the patient's immune response. PMID- 12774015 TI - Triple therapy with amantadine in treatment-naive patients with chronic hepatitis C: a placebo-controlled trial. AB - The antiviral efficacy of amantadine in patients with chronic hepatitis C is controversial. In this randomized, prospective, placebo-controlled, multicenter trial, triple therapy with interferon alfa (IFN-alpha)-2a plus ribavirin and amantadine (amantadine group) was compared with combination therapy IFN-alpha plus ribavirin (control group). Four hundred previously untreated patients with histologically proven chronic hepatitis C were randomly allocated to treatment with amantadine sulphate (100 mg twice daily orally) or a matched placebo together with IFN-alpha induction plus ribavirin (1,000-1,200 mg/day orally) for 48 weeks. The primary end point was sustained virologic response (SVR) defined as undetectable serum hepatitis C virus (HCV) RNA (<100 copies/mL) 24 weeks after the end of treatment. SVR was observed in 52% of the amantadine group and in 43.5% of the control group (P =.11). Among patients with HCV genotype 1 infection, the corresponding SVR rates were 39% and 31%, respectively. The virologic on-treatment response rate in week 24 was significantly higher in the amantadine group as compared with the control group (70% vs. 59%, respectively, P =.016). This beneficial effect was mainly related to HCV type 1-infected patients (63% vs. 47%, respectively, P =.012). Independent factors associated with SVR, according to multiple logistic regression analysis, were amantadine treatment, low baseline HCV RNA, platelet counts (>/=250/nL), pretreatment ALT quotient >/=3, and GGT level (<28 U/L) as well as HCV genotypes other than 1. In conclusion, although we could not demonstrate a significant advantage of the triple regimen in univariate analysis, multivariate analysis offers arguments that amantadine should be considered as a potential anti-HCV drug in future studies. PMID- 12774016 TI - A multicenter study of recombinant human interleukin 12 for the treatment of chronic hepatitis C virus infection in patients nonresponsive to previous therapy. AB - Recombinant human interleukin 12 (IL-12) is an immunomodulatory cytokine that is active against several viruses. Treatment options in patients with chronic hepatitis C with nonresponse to interferon (IFN)-based therapy are limited. Prior dose-ranging studies have indicated drug tolerability and transient suppression of hepatitis C virus (HCV) RNA by IL-12. The aim of this study was to determine the safety and efficacy of prolonged IL-12 therapy in patients who have failed treatment with IFN-alpha +/- ribavirin. A total of 225 patients at 21 U.S. sites who had a history of nonresponse to IFN-alpha or combination IFN-alpha plus ribavirin for treatment of HCV were randomized to 500 ng/kg IL-12 or placebo subcutaneously twice weekly for 12 weeks. The groups were then unblinded; patients receiving IL-12 continued for another 36 weeks, and the placebo group received 48 weeks of treatment with IL-12 in an open-label fashion. HCV RNA, serum alanine aminotransferase (ALT) level, and a repeat liver biopsy were assessed at 24 weeks following therapy. Approximately 1% (2 of 160) of nonresponsive patients enrolled for treatment had a sustained virologic response to IL-12 therapy, but 3% (7 of 225) developed severe adverse events probably related to treatment, resulting in early termination of the trial. Common adverse effects reported by most patients included chills, fever, fatigue, headache, and arthralgia. At termination of the study, 160 patients had received at least 8 weeks of treatment with IL-12. Paired liver biopsy specimens were available for evaluation in 54 patients, but there were no significant changes in Knodell fibrosis or histologic activity index (HAI) scores. In conclusion, IL-12 as monotherapy at the doses used in this trial for chronic hepatitis C has low efficacy, was poorly tolerated, and is unlikely to provide an alternative to conventional IFN-based therapy. PMID- 12774017 TI - Role of the hepatocyte nuclear factor 4alpha in control of the pregnane X receptor during fetal liver development. AB - The fetal liver, the major site of hematopoiesis during embryonic development, acquires additional functions near birth. Among the important liver functions is the response to xenobiotic exposure due to expression of several cytochromes P450 (CYP) and drug efflux transporters. Expression of these genes is regulated by nuclear receptors such as the pregnane X receptor (PXR). In this study, regulation of xenobiotic responses during fetal liver development was analyzed using a fetal hepatocyte primary culture system derived from embryonic day 15 (E15) livers. Hepatocyte nuclear factor (HNF) 4alpha regulates the expression of many genes preferentially in the liver. Expression of several xenobiotic response genes as well as HNF4alpha was increased in fetal hepatocytes stimulated by the hepatic maturation factors oncostatin M (OSM) and Matrigel. To determine the contribution of HNF4alpha to xenobiotic responses in the fetal liver, fetal hepatocytes containing floxed HNF4alpha alleles were cultured and the HNF4alpha gene was inactivated by infection with an adenovirus containing the Cre gene. Expression of CYP3A11 and PXR was suppressed by inactivation of HNF4alpha. An HNF4alpha binding site was characterized in the PXR promoter and found to be required for activation of the PXR promoter in fetal hepatocytes. In conclusion, HNF4alpha is the key transcription factor regulating responses to xenobiotics through activation of the PXR gene during fetal liver development. PMID- 12774018 TI - Enrichment of hepatic progenitor cells from adult mouse liver. AB - Hepatic progenitor cells (HPCs) have been characterized in several drug-treated rodent models and in the fetal liver; however, their properties have not been fully clarified in the normal adult liver, presumably because of their relatively small population and the existence of mature hepatocytes. In an attempt to resolve this issue, we developed a new enrichment system for HPCs using their cell aggregate formation properties. Nonparenchymal cells (NPCs) derived from enzymatically digested liver cells in normal adult mouse liver were treated in a hypoxic 2-hour suspension culture under constant shaking. This procedure resulted in cell aggregate formation and almost complete elimination of mature hepatocytes. Cell aggregates were formed only in Ca(2+)-containing medium, suggesting cadherin-dependent cell-cell adhesion. In these cell aggregates, 95% consisted of vascular endothelial cells that expressed VE-cadherin. The remaining 5% consisted of rapidly proliferating, small epithelial cells that expressed alpha-fetoprotein (AFP), E-cadherin, and albumin but not cytokeratin 19 (CK19), alpha-smooth muscle actin, or VE-cadherin. These results are consistent with an immature hepatic cell phenotype. When these immature hepatic cells were cultured with 10(-7) mol/L dexamethasone and 1% dimethyl sulfoxide, the de novo expression of mature hepatocyte markers such as tryptophan-2,3-dioxygenase (TO) was induced concomitantly with the induction of morphologic characteristics such as mitochondria- and peroxisome-rich cytoplasm and bile canaliculi formation. In conclusion, our methodology allows the enrichment of immature hepatic cells from the normal adult mouse. These cells are capable of growth and maturation along the hepatocyte lineage, indicating that these cells are HPCs. PMID- 12774019 TI - Proteasome inhibition potentiates CYP2E1-mediated toxicity in HepG2 cells. AB - Chronic ethanol consumption causes increased oxidative damage in the liver. Induction of CYP2E1 is one pathway involved in how ethanol produces oxidative stress. Ethanol can cause protein accumulation, decreased proteolysis, and decreased proteasome activity. The objective of this study was to investigate the effect of inhibition of the proteasome activity on CYP2E1-dependent toxicity. HepG2 cells over-expressing CYP2E1 (E47 cells) were treated with arachidonic acid (AA) plus iron, agents important in development of alcoholic liver injury and which are toxic to E47 cells by a mechanism dependent on CYP2E1, oxidative stress, and lipid peroxidation. Addition of various proteasome inhibitors was associated with significant potentiation of the loss of cell viability caused by AA plus iron. Potentiation of toxicity was associated with increased oxidative damage as reflected by an increase in lipid peroxidation and accumulation of oxidized and nitrated proteins in E47 cells and an enhanced decline in mitochondrial membrane potential. Antioxidants prevented the loss of viability and the potentiation of this loss of viability by proteasome inhibition. CYP2E1 levels were elevated about 3-fold by the proteasome inhibitors. Inhibition of proteasome activity also potentiated toxicity of AA alone and toxicity after treatment to remove glutathione (GSH). Similar results were found in hepatocytes from pyrazole-treated rats with high levels of CYP2E1. In conclusion, proteasome activity plays an important role in modulating CYP2E1-mediated toxicity in HepG2 cells by regulating CYP2E1 levels and by removal of oxidized proteins. Such interactions may be important in CYP2E1-catalyzed toxicity of hepatotoxins and in alcohol-induced liver injury. PMID- 12774020 TI - Oxidant-induced hepatocyte injury from menadione is regulated by ERK and AP-1 signaling. AB - Oxidative stress has been implicated as a mechanism for a variety of forms of liver injury. Although reactive oxygen species (ROS) may damage cellular macromolecules directly, oxidant-induced cell death may result from redox effects on signal transduction pathways. To understand the mechanisms of hepatocyte death from oxidative stress, the functions of the mitogen-activated protein kinases (MAPKs) were determined during oxidant-induced hepatocyte injury from menadione. Low, nontoxic, and high toxic concentrations of the superoxide generator menadione were established in the RALA255-10G rat hepatocyte cell line. Death from menadione was blocked by catalase and ebselen, indicating that death was secondary to oxidant generation and not arylation. Treatment with a nontoxic menadione concentration resulted in a brief activation of extracellular signal regulated kinase (ERK) and c-Jun N-terminal kinase (JNK). In contrast, treatment with a toxic menadione concentration induced a prolonged activation of both ERK and JNK. Chemical inhibition of ERK function sensitized RALA hepatocytes to death from previously nontoxic menadione concentrations in association with sustained JNK activation. Adenoviral expression of a dominant-negative protein for c-Jun, a downstream substrate for JNK, blocked death from menadione. The pro-apoptotic effect of c-Jun was not mediated through the mitochondrial death pathway. In conclusion, RALA hepatocyte resistance to oxidant-induced death from menadione is dependent on ERK, whereas cell death is mediated by AP-1 activation. These findings identify signaling pathways that may be therapeutic targets in the prevention or treatment of oxidant-induced liver injury. PMID- 12774021 TI - Elevated hepatocyte levels of the Forkhead box A2 (HNF-3beta) transcription factor cause postnatal steatosis and mitochondrial damage. AB - The Forkhead box (Fox) transcription factor Foxa2 (HNF-3beta) and related family members Foxa1 (HNF-3alpha) and Foxa3 (HNF-3gamma) act in concert with other hepatocyte nuclear factors (HNF) to coordinately regulate liver-specific gene expression. To circumvent the hepatic functional redundancy of the Foxa proteins, we used the T-77 transgenic (TG) mouse line in which the -3-kb transthyretin (TTR) promoter functioned to increase hepatocyte expression of the Foxa2 cDNA. Adult TG mice exhibited reduced hepatic glycogen and progressive liver injury, but maintained normal serum levels of glucose, insulin, and glucagon. In this study, we further characterized the postnatal liver defect in TTR-FoxA2 TG mice. The postnatal TG mice displayed significant reduction in serum glucose levels and in hepatocyte glycogen storage without increased serum levels of ketone bodies and free fatty acid suggesting that they are not undergoing a starvation response. We show that TG liver developed a substantial transient steatosis, which reached a maximum at postnatal day 5 and is associated with increased expression of hepatic genes involved in fatty acid and triglyceride synthesis, lipid beta-oxidation, and amino acid biosynthesis. Furthermore, transmission electron microscopy analysis of postnatal TG liver revealed extensive mitochondrial membrane damage, which is likely due to reactive oxygen species generated from lipid beta-oxidation. In conclusion, our model proposes that in response to reduction in hepatocyte glycogen storage, the TTR-Foxa2 TG mice survive by maintaining sufficient serum levels of glucose through gluconeogenesis using deaminated amino acids with dicarboxylate products of peroxisomal lipid beta-oxidation shuttled through the tricarboxylic acid cycle. PMID- 12774022 TI - Mechanisms for sensitization to TNF-induced apoptosis by acute glutathione depletion in murine hepatocytes. AB - We previously reported that depletion of glutathione in murine hepatocytes by diethylmaleate (DEM) or acetaminophen (APAP) leads to oxidative stress-dependent necrosis and sensitizes to tumor necrosis factor (TNF)-induced apoptosis in an oxidative stress-independent fashion, which could not be explained by interference with nuclear factor kappaB (NF-kappaB) nuclear translocation. The present report explores the mechanisms of these effects. We observed that DEM led to necrosis when both mitochondrial and cytosol glutathione were depleted profoundly but sensitized to TNF-induced apoptosis when cytosol glutathione was depleted selectively. DEM and APAP lead to a significant decrease in reduced glutathione (GSH)/glutathione disulfide (GSSG) ratio. Glutathione depletion by DEM or APAP was associated with inhibition of TNF-induced NF-kappaB transactivation of anti-apoptotic genes, including inducible nitric oxide synthase (i-NOS). Provision of exogenous NO partially abrogated the sensitization to TNF in response to glutathione depletion. Glutathione depletion alone led to sustained increase in phospho-jun levels and c-Jun-N-terminal kinase (JNK) activity. JNK inhibitor partially blocked the sensitization to TNF-induced apoptosis accompanying glutathione depletion. In conclusion, these findings suggest that extramitochondrial glutathione depletion alters the thiol-disulfide redox state, leading to inhibition of NF-kappaB transactivation of survival genes and to sustained activation of JNK, both of which contribute to the sensitization to TNF-induced apoptosis. PMID- 12774023 TI - Glucagon induces the plasma membrane insertion of functional aquaporin-8 water channels in isolated rat hepatocytes. AB - Although glucagon is known to stimulate the cyclic adenosine monophosphate (cAMP) mediated hepatocyte bile secretion, the precise mechanisms accounting for this choleretic effect are unknown. We recently reported that hepatocytes express the water channel aquaporin-8 (AQP8), which is located primarily in intracellular vesicles, and its relocalization to plasma membranes can be induced with dibutyryl cAMP. In this study, we tested the hypothesis that glucagon induces the trafficking of AQP8 to the hepatocyte plasma membrane and thus increases membrane water permeability. Immunoblotting analysis in subcellular fractions from isolated rat hepatocytes indicated that glucagon caused a significant, dose dependent increase in the amount of AQP8 in plasma membranes (e.g., 102% with 1 micromol/L glucagon) and a simultaneous decrease in intracellular membranes (e.g., 38% with 1 micromol/L glucagon). Confocal immunofluorescence microscopy in cultured hepatocytes confirmed the glucagon-induced redistribution of AQP8 from intracellular vesicles to plasma membrane. Polarized hepatocyte couplets showed that this redistribution was specifically to the canalicular domain. Glucagon also significantly increased hepatocyte membrane water permeability by about 70%, which was inhibited by the water channel blocker dimethyl sulfoxide (DMSO). The inhibitors of protein kinase A, H-89, and PKI, as well as the microtubule blocker colchicine, prevented the glucagon effect on both AQP8 redistribution to hepatocyte surface and cell membrane water permeability. In conclusion, our data suggest that glucagon induces the protein kinase A and microtubule-dependent translocation of AQP8 water channels to the hepatocyte canalicular plasma membrane, which in turn leads to an increase in membrane water permeability. These findings provide evidence supporting the molecular mechanisms of glucagon induced hepatocyte bile secretion. PMID- 12774024 TI - Hydrophilic but not hydrophobic bile acids prevent gallbladder muscle dysfunction in acute cholecystitis. AB - The pathogenesis of acute cholecystitis (AC) is controversial. Bile acids may be involved in the pathogenesis of AC because the hydrophobic chenodeoxycholic acid (CDCA) reproduced in vitro the muscle dysfunction observed in AC and was prevented by the hydrophilic ursodeoxycholic acid (UDCA). The present study examined the in vivo effects of UDCA or CDCA on gallbladder muscle dysfunction caused by AC. Guinea pigs were treated with placebo, UDCA, or CDCA for 2 weeks before sham operation or induction of AC by bile duct ligation (BDL) for 3 days. Pretreatment with oral UDCA prevented the defective contraction in response to agonists (acetylcholine [ACh], cholecystokinin 8 [CCK-8], and KCl) that occurs after BDL. Prostaglandin (PG) E(2)-induced contraction remained normal in the placebo and UDCA-treated groups but was impaired in the CDCA-treated group. Treatment with UDCA also prevented the expected increase in the levels of H(2)O(2), lipid peroxidation, and PGE(2) content in the placebo-treated AC group, whereas CDCA caused further increases in these oxidative stress markers. The binding capacity of PGE(2) to its receptors and the activity of catalase were reduced after treatment with CDCA. Treatment with UDCA enriched gallbladder bile acids with its conjugates and reduced the percentage of CDCA conjugates. In contrast, treatment with CDCA significantly decreased the percentage of UDCA in bile. In conclusion, oral treatment with UDCA prevents gallbladder muscle damage caused by BDL, whereas oral treatment with CDCA worsens the defective muscle contractility and the oxidative stress. PMID- 12774025 TI - Long-term expression of angiostatin suppresses metastatic liver cancer in mice. AB - Metastatic liver cancer has a very poor prognosis and lacks effective therapy. Anti-angiogenic therapies, which starve tumors of blood supply, have proven to be effective in preclinical models because tumor growth is angiogenesis dependent. However, long-term, high-level, and sustained expression of angiogenesis inhibitors, such as angiostatin, is necessary to prevent dormant tumors from becoming active again. To achieve this objective, we engineered a recombinant adeno-associated virus (AAV) vector encoding mouse angiostatin, an endogenous inhibitor of tumor vascularization. After intraportal delivery of this vector, high-level, stable transgene expression of angiostatin lasting for at least 6 months was observed locally in hepatocytes. Gene transfer of AAV-angiostatin via the portal vein led to significant suppression of the growth of both nodular and metastatic EL-4 lymphoma tumors established in the liver and prolonged the survival time of the mice. The growth of neovessels was inhibited significantly, and extensive apoptosis of tumor cells was observed. The anti-angiogenic activity of angiostatin was independent of vascular endothelial growth factor (VEGF). The AAV-angiostatin viruses did not appear to be toxic to mice, and there was no detectable apoptosis of hepatocytes. In conclusion, these encouraging results warrant future investigation of the use of AAV-mediated anti-angiogenic gene therapy for targeting unresectable liver metastases, especially after surgical removal of primary tumors. PMID- 12774026 TI - Insulin receptor substrate-4 signaling in quiescent rat hepatocytes and in regenerating rat liver. AB - This study was designed to characterize insulin receptor substrate-4 (IRS-4) in isolated rat hepatocytes and to examine its role in liver regeneration. Subcellular fractionation revealed that 85% of IRS-4 is located at isolated hepatocyte plasma membranes. The distribution of IRS-4 among intracellular compartments remained unchanged in insulin-stimulated cells. Two bands corresponding to 145 and 138 kd were observed in immunoblotting experiments. Immunoprecipitation of hepatocyte lysates with a highly specific antibody against IRS-4 led to an insulin and insulin-like growth factor 1 (IGF-1)-dependent increase in phosphotyrosine residues of the 145-kd band. IRS-4 was found to be associated with Src homology 2 (SH2) domain-containing proteins (phosphatidylinositol 3-kinase [PI 3-kinase] and Src homology phosphatase [SHP 2]) and with protein kinase C zeta (PKC zeta). Insulin and IGF-1 elicited a rapid and dose-dependent binding of these 3 proteins to IRS-4. These data suggest that IRS-4 is insulin-/IGF-1-activated by phosphorylation and not by translocation, inducing the recruitment of SH2 domain-containing proteins and PKC zeta to the membrane. To evaluate the possible role of IRS-4 in liver regeneration, we also examined this system after partial hepatectomy (PH). One day after PH, IRS-1 expression increased, consistent with a stimulatory role in the regenerative process, whereas it decreased 7 days after liver resection. This drastic IRS-1 depletion occurred at the expense of increased IRS-2 and IRS-4 expression 7 days after PH. In addition, at this period of time after surgery, the in vivo insulin stimulation of remnant rat livers showed an increase in IRS-4/PI 3-kinase association. Given that 1 and 7 days after PH isolated hepatocytes responded similarly to insulin in terms of induced cell proliferation, a compensatory role is proposed for IRS-2/4 induction. In conclusion, IRS-4 is activated by insulin and IGF-1-like IRS-1 in rat hepatocytes, and the induced expression of IRS-4 is a compensatory mechanism that plays a role in conditions of liver regeneration. PMID- 12774027 TI - A practice guideline on Wilson disease. PMID- 12774028 TI - Screening tests for hepatocellular carcinoma. PMID- 12774029 TI - Renal sodium retention in portal hypertension and hepatorenal reflex: from practice to science. PMID- 12774030 TI - Prediction of spontaneous viral clearance in acute hepatitis C by viral load measurements. PMID- 12774031 TI - Ammonia metabolism in cirrhotic patients with TIPS. PMID- 12774032 TI - Management of HCV-infected health care workers. PMID- 12774033 TI - The way to a man's heart is through his liver. PMID- 12774034 TI - Genetic linkage and imprinting effects on body mass index in children and young adults. AB - Body mass index (BMI) is used as a measure of fatness. Here we performed a genome wide scan for genes related to BMI, while allowing for the possible effects of imprinting. We applied a sib pair linkage analysis to a sample of primarily children and young adults by using the Haseman-Elston method, which we modified to model the separate effects of paternally and maternally derived genetic factors. After stratification of sib pairs according to age, a number of regions showing linkage with BMI were identified. Most linkage and imprinting effects were found in children 5-11 years of age. Strongest evidences for linkage in children were found on chromosome 20 at 20p11.2-pter near the marker D20S851 (LOD(Total)=4.08, P=0.000046) and near the marker D20S482 (LOD(Total) =3.55, P=0.00016), and Chromosome 16 at 16p13 near the marker ATA41E04 (LOD(Total) =3.12, P=0.00025), and those loci did not show significant evidence for imprinting. Six regions showing evidence of imprinting were 3p23-p24 (paternal expression), 4q31.1-q32 (maternal expression), 10p14-q11 (paternal expression), and 12p12-pter (paternal expression) in children, and 4q31-qter (paternal expression) and 8p (paternal expression) in adults. PMID- 12774035 TI - A gene for familial isolated chronic nail candidiasis maps to chromosome 11p12 q12.1. AB - Chronic mucocutaneous candidiases (CMC) are a group of rare disorders where an altered immune response against Candida leads to persistent and/or recurrent infections of the skin, nails, and mucous membranes. We analysed a five generation Italian family with an isolated form of CMC, affecting nails only, in the presence of low serum concentration of intercellular adhesion molecule I (ICAM-1). We excluded linkage to candidate regions on chromosomes 2p (CMC with thyroid disease), 21q22.3 (APECED), and 19q13 (ICAM-1). We then carried out a genome-wide scan and assigned the CMC locus to a 19 cM pericentromeric region on chromosome 11. PMID- 12774036 TI - Independent effects of the -219 G>T and epsilon 2/ epsilon 3/ epsilon 4 polymorphisms in the apolipoprotein E gene on coronary artery disease: the Southampton Atherosclerosis Study. AB - A number of studies have shown that coronary artery disease severity is associated with the epsilon 2/ epsilon 3/ epsilon 4 polymorphism in the coding region of the apolipoprotein E gene. In this study, we investigated whether the severity of the disease was also influenced by a functional polymorphism (-219 G>T) in the promoter of the gene, and if so, whether the effects of the two polymorphisms were independent. A cohort of 1170 patients with angiographically documented coronary artery disease were genotyped for the two polymorphisms. The frequency of the epsilon 4 allele of the epsilon 2/ epsilon 3/ epsilon 4 polymorphism increased linearly with increasing number of diseased vessels, so did the -219T allele of the -219 G>T polymorphism. In the sample as a whole, logistic regression analyses indicated that compared with the G/G genotype, the T/T genotype conferred an odds ratio of 1.598 (95% CI=1.161-2.201, P=0.004) in favor of increased disease severity, and the relationship remained significant after adjustment for epsilon 2/ epsilon 3/ epsilon 4 polymorphism genotypes, plasma cholesterol and triglyceride levels, and other risk factors. The effect of the T/T genotype on disease severity was more significant in patients who did not carry the epsilon 4 allele (OR=1.510, 95% CI=1.028-2.221) than in epsilon 4 allele carriers (OR=1.303, 95% CI=0.619-2.742). There was considerable linkage disequilibrium between the two polymorphisms (rho=0.9, P<0.001). Logistic regression analysis showed that the -219T- epsilon 4 haplotype conferred an odds ratio of 1.488 (95% CI=1.133-1.954). These findings suggest that the -219 G>T and epsilon 2/ epsilon 3/ epsilon 4 polymorphisms, which may affect respectively the quantity and quality of apoE, have independent and possibly additive effects on coronary artery disease severity. PMID- 12774037 TI - Apolipoprotein B 3'-VNTR polymorphism in Eastern European populations. AB - Apolipoprotein B 3' (3' ApoB) minisatellite polymorphism was studied in healthy unrelated individuals from the Russian Federation and the Republic of Belarus, in 10 populations from five ethnic groups: Russians, Byelorussians, Adygeis, Kalmyks and Yakuts. The analysis was carried out using PCR and electrophoresis followed by silver staining. Overall, 25 alleles of the 3' ApoB minisatellite, ranging from 25 to 55 repeats, were detected. Heterozygosity indices were high and varied from 0.73 to 0.84. The distributions of alleles of this minisatellite in the Caucasoid populations (Russians, Byelorussians and Adygeis) had a bimodal character, whereas that for Mongoloid populations (Kalmyks and Yakuts) had a unimodal distribution. Nei's genetic distances between the populations studied and some reference populations of Europe and Asia were estimated. Despite their allele distribution homogeneity, different East Slavonic ethnic groups were clearly resolved by multidimensional analyses. The East Slavonic and Adygei populations revealed a high similarity with European Caucasoids. The Mongoloid populations (Kalmyks and Yakuts) were considerably different from those of the European Caucasoid populations, but were similar to other Asian Mongoloid populations. The results demonstrate the variability of 3' ApoB minisatellite polymorphism not only in distant populations but also, to a certain extent, in genetically relative ones. PMID- 12774038 TI - CGH and direct diagnosis of mosaic structural chromosomal abnormalities: description of a mosaic ring chromosome 17 and review of the literature. AB - We report the characterisation of a de novo supernumerary chromosome marker in a mosaic state (50%) by comparative genomic hybridisation (CGH) in an 8-year-old child with hypotonia, dysmorphia and mild-to-moderate mental retardation. We describe the combined use of CGH and fluorescence in situ hybridisation (FISH) to identify the origin of the additional chromosomal material. Visual analysis of 10 CGH-metaphase spreads revealed a gain of green fluorescent signal on pericentromeric region of chromosome 17. The CGH finding was confirmed by FISH analysis using a whole chromosome 17 paint, a chromosome 17 centromeric probe and the probe coding for the Smith-Magenis locus in 17p11.2. These results show that performing both CGH and FISH in combination with classical karyotyping will certainly allow the identification of imbalanced chromosome rearrangements and, by the way, allow the identification of genes involved in mental retardation and/or malformative pathology. PMID- 12774039 TI - Clinical and molecular analysis of nine families with Adams-Oliver syndrome. AB - Adams-Oliver syndrome (AOS) is defined by the combination of limb abnormalities and scalp defects, often accompanied by skull ossification defects. We studied nine families affected with AOS, eight of which have not been clinically described before. In our patients, scalp abnormalities were most often found, followed by limb and skull defects. The most common limb abnormalities appeared to be brachydactyly, syndactyly of toes 2 and 3 and hypoplastic toenails. Additional features observed were cutis marmorata telangiectatica congenita, cryptorchidism and cardiac abnormalities. In an attempt to identify the disease causing mutations in our families, we selected two genes, ALX4 and MSX2, which were considered serious candidates based on their known function in skull and limb development. Mutation analysis of both genes, performed by direct sequencing, identified several polymorphisms, but no disease-causing mutations. Therefore, we can conclude that the AOS in our set of patients is not caused by mutations in ALX4 or MSX2. PMID- 12774040 TI - Similar contributions of BRCA1 and BRCA2 germline mutations to early-onset breast cancer in Germany. AB - This study was undertaken to investigate the prevalence of BRCA1 and BRCA2 germline mutations in 91 German patients unselected for family history, who were diagnosed with breast cancer before the age of 41 years. Clinical information and blood samples were obtained from all patients. A comprehensive BRCA1 and BRCA2 mutational analysis was performed using the protein truncation assay and single strand conformational polymorphism analysis followed by DNA sequencing of variant signals detected by these assays. Five different deleterious germline mutations including four frameshift mutations and one missense mutation were identified, three in BRCA1 (3.3%) and two mutations (2.2%) in BRCA2. Both BRCA2 mutations are novel and might be specific for the German population. An additional BRCA1 missense mutation previously described and classified as an unknown variant was found. This mutation was also detected in two breast cancer patients of family P 328 and not in 140 healthy controls suggesting that it is disease associated. In addition, one common polymorphism and five novel intronic sequence variants with unknown significance were found. Our findings show that mutations in BRCA1 and BRCA2 may contribute similarly to early-onset breast cancer in Germany. Given current constraints on health-care resources, these results support the notion that BRCA1 and BRCA2 mutation screening may have the strongest impact on health care when targeted to high-risk populations. PMID- 12774041 TI - Mutations in PAX1 may be associated with Klippel-Feil syndrome. AB - Pax genes are a highly conserved family of developmental control genes that encode transcription factors. In vertebrates, Pax genes play a role in pattern formation during embryogenesis. Mutations in Pax genes have been associated with both spontaneous mouse mutants and congenital human diseases. The mouse Pax1 mutant phenotype undulated is characterised by vertebral segmentation defects reminiscent of the human disorder Klippel-Feil syndrome (KFS). To determine whether PAX1 haploinsufficiency plays a role in KFS, we have defined the gene structure of the human PAX1 gene and screened 63 KFS patients for mutations in this gene. Differences in the PAX1 sequence were detected in eight patients. Two patients had a silent change within the paired box that was also seen in 2/303 control chromosomes. The other variants were missense, silent or intronic changes not represented in the control panel tested. The significance of these results and the possible role of PAX1 in the pathogenesis of KFS are discussed. PMID- 12774042 TI - An empirical survey on biobanking of human genetic material and data in six EU countries. AB - Biobanks correspond to different situations: research and technological development, medical diagnosis or therapeutic activities. Their status is not clearly defined. We aimed to investigate human biobanking in Europe, particularly in relation to organisational, economic and ethical issues in various national contexts. Data from a survey in six EU countries (France, Germany, the Netherlands, Portugal, Spain and the UK) were collected as part of a European Research Project examining human and non-human biobanking (EUROGENBANK, coordinated by Professor JC Galloux). A total of 147 institutions concerned with biobanking of human samples and data were investigated by questionnaires and interviews. Most institutions surveyed belong to the public or private non-profit making sectors, which have a key role in biobanking. This activity is increasing in all countries because few samples are discarded and genetic research is proliferating. Collections vary in size, many being small and only a few very large. Their purpose is often research, or research and healthcare, mostly in the context of disease studies. A specific budget is very rarely allocated to biobanking and costs are not often evaluated. Samples are usually provided free of charge and gifts and exchanges are the common rule. Good practice guidelines are generally followed and quality controls are performed but quality procedures are not always clearly explained. Associated data are usually computerised (identified or identifiable samples). Biobankers generally favour centralisation of data rather than of samples. Legal and ethical harmonisation within Europe is considered likely to facilitate international collaboration. We propose a series of recommendations and suggestions arising from the EUROGENBANK project. PMID- 12774043 TI - Over-representation of two specific haplotypes among chromosomes harbouring BRCA1 mutations. AB - The BRCA1 gene is included in a 200-400 kb region that is subjected to a recombination suppression mechanism; this region shows nearly complete linkage disequilibrium for a series of common biallelic polymorphisms, all of them with rarer allele frequency close to 0.4. These series of SNPs define two major haplotypes designated as class I and class II. In the present study, we have determined haplotype classes in the index case of 106 breast/ovarian cancer families previously screened for mutations in the BRCA genes and we have found that haplotype II (the less frequent in the control population) is over represented among chromosomes harbouring mutations in BRCA1. In addition, we have defined a subtype of chromosomes characterized by haplotype I and one specific allele for the microsatellite marker D17S855, which are also more frequently associated with BRCA1 mutations. These findings may have important consequences for the selection of families with higher probabilities of carrying mutations in the BRCA1 gene. PMID- 12774044 TI - Why aren't we performing more allografts for aggressive non-Hodgkin's lymphoma? AB - Allogeneic stem cell transplantation has an under-appreciated role in the management of intermediate-grade non-Hodgkin's lymphoma. It provides several advantages over autologous stem cell transplantation including provision of a lymphoma-free graft, reduced rates of secondary myelodysplastic syndrome and leukemia, and a potentially curative graft-versus-lymphoma effect. When applied to chemosensitive patients, the lower relapse rates and reasonable long-term outcomes make allogeneic transplantation a promising therapy to pursue. Patient populations, such as those with bone marrow involvement or very high-risk disease, can be identified as having suboptimal outcomes after autotransplantation and may benefit from such an approach. While the exact role of allogeneic stem cell transplantation remains to be determined, broad recommendations can be suggested for the management of patients with intermediate grade lymphoma. New approaches to allogeneic transplantation, including the use of matched-unrelated donors and reduced-intensity conditioning regimens, may expand the applicability of this potentially curative modality. PMID- 12774045 TI - Impact of obesity in the setting of high-dose chemotherapy. AB - Obesity is a common and increasing problem affecting the developed world. Chemotherapy pharmacokinetics in the context of obesity demonstrate prolonged elimination time for several agents, but there is no consensus regarding optimal dosing schemes. Retrospective studies suggest differences in toxicity for obese adult patients but these data may be greatly impacted by chemotherapy dose adjustments. Outcome data from several studies have suggested that obese adult patients suffer poorer disease-free survival and overall survival, although this is not a uniform conclusion in all studies. Prospective studies of chemotherapy pharmacokinetics are needed to address the issue of optimal chemotherapy dosing in the obese population. PMID- 12774046 TI - Impact of transplanted CD34+ cell dose in allogeneic unmanipulated peripheral blood stem cell transplantation. AB - The impact of the CD34+ cell dose on chronic graft-versus-host disease (cGVHD) and the clinical outcome was analyzed in 41 consecutive adult patients submitted to allogeneic peripheral blood stem cell transplantation from HLA-identical siblings. The patients were classified into 'low' or 'high' CD34+ cell dose groups based on whether they received less or more than a median CD34+ cell dose of 10.5 x 10(6)/kg, respectively. There was a significant difference in the incidence of extensive cGVHD (low vs high group, 25.0 vs 66.7%, P=0.021) and relapse (47.6 vs 20.0%, P=0.049) between the two groups. With a median follow-up of 335 days, the 3-year survival estimate for the whole population was 47.9%, while that for the low and high groups was 29.9 and 67.8%, respectively (P=0.0434). An inverse relation was noted between the relapse rate and the incidence of extensive cGVHD (P=0.043). It would appear reasonable that the optimal dose of CD34+ cells should be determined based on the disease status or aggressiveness of the malignant cells in each patient. Yet, in the case of patients with a high risk of relapse, transplantation with a CD34+ cell dose of >10.5 x 10(6)/kg would seem to be acceptable to minimize the risk of relapse. PMID- 12774047 TI - Myeloablative intensified conditioning regimen with in vivo T-cell depletion (ATG) followed by allografting in patients with advanced multiple myeloma. A phase I/II study of the German Study-group Multiple Myeloma (DSMM). AB - We investigated toxicity and efficacy of in vivo T-cell depletion with anti thymocyte globulin (ATG) as part of an intensified myeloablative conditioning regimen followed by allogeneic stem cell transplantation in patients with advanced multiple myeloma. The conditioning regimen consisted of modified total body irradiation, busulfan and cyclophosphamide (n=15) or in the case of prior dose-limiting radiotherapy of busulfan and cyclophosphamide (n=3). The median age was 44 years (range, 29-53) and the median time from diagnosis to transplant was 12 months (range, 6-144). Grade II-IV acute graft-versus-host disease (GvHD) occurred in six patients (35%). Severe grade III/IV GvHD developed in one patient (6%). Three patients died of therapy-related causes (17%). A complete remission (CR) with negative immunofixation after allogeneic transplantation was seen in eight of the evaluable patients (53%). After a median follow-up of 41 months (range, 8-84), the estimated overall survival at 6 years for all patients is 77% (CI 95%: 58-96%). The estimated progression-free survival at 6 years for all patients is 31% (CI 95%: 2-59%) and 46% (CI 95%: 9-83%) for patients with CR. In vivo T-cell depletion with ATG resulted in a low rate of severe GvHD with low treatment-related mortality, and a substantial number of long-term survivors. PMID- 12774048 TI - Allogeneic bone marrow transplantation for children with histiocytic disorders: use of TBI and omission of etoposide in the conditioning regimen. AB - The histiocytoses are rare disorders of antigen-processing phagocytic or antigen presenting cells. Allogeneic bone marrow transplantation (BMT) can be curative of these disorders. We report a series of five children with Langerhans cell histiocytosis (n=2) or hemophagocytic lymphohistiocytosis (n=3), who received allogeneic BMT with a total body irradiation (TBI)-containing regimen (TBI, cytarabine, and cyclophosphamide) at our institution between 1995 and 2000. One of these patients received busulfan, cyclophosphamide, and etoposide for the first of two BMTs. All grafts except one (a matched sibling-donor graft) were T cell-depleted grafts from unrelated donors. All received cyclosporine graft versus-host disease (GvHD) prophylaxis; the recipient of the matched sibling graft also received methotrexate. Three patients engrafted at a median of 24 days after transplantation. The patient who did not receive TBI experienced primary graft failure and recurrent disease. After the TBI-containing conditioning regimen was given, a second transplant engrafted on day +17. One patient with concurrent myelodysplastic syndrome died of toxicity on day +33 without evidence of engraftment. No acute or chronic GvHD was observed. Four patients survive disease-free, a median of 63 months after transplantation, all with Lansky performance scores of 100. We conclude that a conditioning regimen containing TBI but not etoposide is effective in allogeneic BMT for children with histiocytic diseases. PMID- 12774049 TI - Low incidence of severe acute graft-versus-host disease in children given haematopoietic stem cell transplantation from unrelated donors prospectively matched for HLA class I and II alleles with high-resolution molecular typing. AB - We evaluated the outcome of 63 children given haematopoietic stem cell transplantation from unrelated donors (URD-HSCT) prospectively selected using DNA high-resolution typing of both HLA class I and class II loci. Thirty patient/donor pairs (48%) were fully matched. Among the others, HSCT was performed in the presence of one (n=22), two (n=9), or three (n=2) HLA disparities. Patients had either malignant (n=46) or non-malignant (n=17) disease. In all cases, graft-versus-host disease (GVHD) prophylaxis consisted of cyclospor-in A, short-term methotrexate and pretransplant anti-thymocyte globulin. The probability of haematopoietic recovery at day 100 was 97%. Two patients experienced primary graft failure. The cumulative probability of grades III-IV acute GVHD and of extensive chronic GVHD equalled 8 and 14%, respectively. A total of 12 patients died of transplant-related complications. The probability of transplant-related mortality (TRM) at 100 and 180 days was 10 and 15%, respectively, whereas the cumulative incidence of TRM was 22%. The probability of GVHD-related mortality equalled 6% at 2.5 years. The overall and disease-free survival rates were 67 and 65%, respectively. URD-HSCT with donor selection based on high-resolution HLA typing is associated with low incidence of both severe acute GVHD and graft failure. The observed outcome is comparable to that of children transplanted from HLA-identical siblings. PMID- 12774050 TI - Outcome of 67 patients with solid tumors relapsed after high-dose chemotherapy and peripheral blood stem cell transplantation. AB - We retrospectively analyzed the outcome of 67 patients with breast (n=24), ovarian (n=11) or testicular cancer (n=32) treated for relapse after high-dose chemotherapy (HDC) and peripheral blood stem cell (PBSC) transplantation. Treatment, survival and toxicity were analyzed. Patients with breast, ovarian or testicular cancer received a mean of 5.9 (range 1-24), 5.1 (1-13) and 4.6 (1-13) regimens for relapse after HDC. Overall response at the end of the observation period was 20.8% for patients with breast cancer (three complete (CR) and two partial responses (PR)), 45.5% (one CR, four PR) for ovarian and 9.4% (three PR) for testicular cancer patients. The mean overall-survival (OAS) from first relapse was 28 (range 3-44), 17 (2-24) and 10 (1-28) months, respectively. Leukocytopenia grade 3/4 occurred in 27-63% of patients, and thrombocytopenia grade 3/4 was observed in 58-88%, respectively. Nonhematological grade 3/4 toxicities were below 20%. In conclusion, patients with relapse after HDC usually have a poor outcome but long-term survivors are observed. Hematological toxicity is common, while other severe side effects are less frequent. PMID- 12774051 TI - The degree of matching at HLA-DPB1 predicts for acute graft-versus-host disease and disease relapse following haematopoietic stem cell transplantation. AB - The importance of matching for HLA-DPB1 in unrelated donor haematopoietic stem cell (HSC) transplantation is little understood. Most transplant centres do not, currently, prospectively match for DPB1, but emerging data show that DPB1 matching does play a role in determining outcome. We studied the impact of HLA DPB1 matching on outcome in 143 recipients of T-cell depletion transplants, who matched with their respective unrelated donors (allelic level) at HLA-A, -B, -C, DRB1 and -DQB1. Of those matched at DPB1, 47.2% (17/36) developed acute graft versus-host disease (aGvHD) as compared to 66.3% (55/83) of those who were mismatched. This led to a 19.1% (95% CI 0.1-38.3%) increase in the chance of developing aGvHD in mismatched patients (P=0.049). Relapse of the original disease occurred in 51 recipients; 23 of 37 (62%) matched at both DPB1 alleles, 28 of 82 (34%) were mismatched at one or two DPB1 alleles. Thus, there was a significantly higher relapse rate (P=0.0011) in transplant recipients who matched at both DPB1 alleles. In conclusion, a donor/recipient DPB1 match was associated with a significantly lower incidence of aGvHD and a significantly higher incidence of disease relapse. This study provides further evidence for an immunogenic role of HLA-DPB1 in HSC transplants. PMID- 12774052 TI - Correlation of early lymphocyte recovery and progression-free survival after autologous stem-cell transplant in patients with Hodgkin's and non-Hodgkin's Lymphoma. AB - The importance of the association between early lymphocyte recovery and outcome has not been well studied in autologous stem cell transplantation (ASCT). In this retrospective study, we analyzed 90 consecutive patients with non-Hodgkin's and Hodgkin's lymphoma who underwent ASCT. Patients were divided into two groups: group 1 with absolute lymphocyte count (ALC) on day +15 below the median of 667/mm(3), and group 2 with ALC >or=667/mm(3). The median progression-free survival (PFS), but not overall survival (OS), was significantly longer in group 2 when compared to group 1 (16 months vs not reached P=0.02). Group 2 patients also had significantly shorter hospital stay, received higher CD34(+) cell dose, and had shorter time to neutrophil recovery. Multivariate analysis demonstrated day +15 ALC to be an independent prognostic indicator for PFS, but not OS, while CD34(+) cell dose and the number of pretransplant treatments were better predictors for both PFS and OS. We conclude that higher day +15 ALC may independently predict better PFS after ASCT for lymphoma patients; however, whether this merely reflects faster overall recovery caused by higher infused CD34(+) cell dose and less pretransplant therapy needs further investigation. PMID- 12774053 TI - High rates of infection and colonization by nontuberculous mycobacteria after allogeneic hematopoietic stem cell transplantation. AB - Nontuberculous mycobacteria (NTM) are essentially ubiquitous and can infect both immunocompetent and immunocompromised hosts. However, NTM infection is surprisingly uncommon in reports from allogeneic hematopoietic stem cell transplant (alloSCT) centers that do not routinely perform allograft T-cell depletion. We reviewed medical records for all adult patients who underwent alloSCT at our center between January 1993 and December 2001. American Thoracic Society and Centers for Disease Control and Prevention guidelines Were used to define definite, probable, and possible NTM infection. Of 571 patients, 36 of 372 (9.7%) T-cell depleted and 14 of 199 (7.0%) conventional alloSCT recipients (P=0.26) had a positive culture for NTM after alloSCT. Of the 50 patients with NTM infection, 16 had definite infection and 34 had probable or possible infection. Rates of NTM infection were 5 to 20-fold higher than rates reported by other centers. Of the 16 definite infections, nine were caused by Mycobacterium haemophilum. Two patients had disseminated M. avium complex (MAC) infection and one had a vascular catheter infected by MAC. Three patients died from complications of NTM infection. Patients with probable or possible NTM infection had markedly different epidemiology, risk factors, site and species of NTM infection, and prognosis than patients with definite NTM infection. PMID- 12774054 TI - Pre-emptive therapy with rituximab for prevention of Epstein-Barr virus associated lymphoproliferative disease after hematopoietic stem cell transplantation. AB - Epstein-Barr virus (EBV)-associated lymphoproliferative disease (LPD) is a life threatening complication following hematopoietic stem cell transplantation (HSCT). Therefore, early diagnosis of EBV reactivation and pre-emptive therapy may be clinically useful. We report three patients who presented with an extremely high EBV load in peripheral blood mononuclear cells and plasma without evidence of EBV disease. Following pre-emptive therapy with a single dose of rituximab, a concordant decrease of EBV-genome copies and B lymphocytes was observed. In all three patients, no EBV-associated LPD occurred. We conclude that pre-emptive therapy with rituximab appears to be effective for prevention of EBV associated LPD after HSCT. PMID- 12774055 TI - IL-2 does not enhance the conversion to complete donor chimerism following nonmyeloablative hematopoietic cell transplantation in dogs. AB - A dog model of stable mixed hematopoietic chimerism was established in which leukocyte-antigen-identical littermates receive nonmyeloablative total body irradiation before hematopoietic cell transplantation and postgrafting immunosuppression with mycophenolate mofetil and cyclosporine. Unmodified donor lymphocyte infusion (DLI) into stable mixed chimeras failed to increase donor chimerism, while DLI from donors sensitized to recipient minor-histocompatibility antigens promptly converted all recipients to complete donor chimerism. This established a model for studying approaches to enhance the graft-versus-host (GVH)-effect, a potential surrogate for graft-versus-leukemia activity. We asked if interleukin-2 (IL-2) given after unmodified DLI could result in reliable conversion to complete donor chimerism. IL-2, 4 x 10(5) IU/kg/day, was administered to six mixed chimeric dogs for 14 days. Four dogs received unmodified DLI with IL-2. At 20-40 weeks after DLI, all dogs remained mixed chimeras. For the two recipients of IL-2 only, mixed chimerism also remained unchanged. These results show that IL-2 given with DLI after nonmyeloablative transplantation in dogs is not effective in reliably converting mixed to complete donor chimerism. PMID- 12774056 TI - Impact of stem cell donation modality on normal donor quality of life: a prospective randomized study. AB - As part of a previously reported trial comparing granulocyte-colony stimulating factor (G-CSF) stimulated bone marrow and peripheral blood stem cells (PBSCs) in allogeneic stem cell transplantation, we included a questionnaire to compare donor morbidity and long-term complications between the two donation procedures. Bone marrow donation was associated with significantly more donors experiencing localized pain at the donation site compared to PBSC collection. However, this was not associated with any increased delay in returning to normal activity. Although a minority of bone marrow donors suffered chronic pain at the donation site, no serious long-term side effects relating to G-CSF stimulated stem cell donation were identified. PMID- 12774057 TI - A rat model of syngeneic bone marrow transplantation during breast cancer therapy. AB - The purpose of this study was to develop a breast cancer model in rats, in which myeloablative chemotherapy and syngeneic bone marrow transplantation (SBMT) could be evaluated systematically for therapeutic effect. The Wistar-Furth (WF) DMBA-4 breast cancer cell line transplanted into naive WF rats produced rapidly growing tumors that were lethal within 2 months. SBMT was performed following preparation with a regimen (Bu-Cy), consisting of busulfan 16 mg/kg by gastric gavage on days -3 and -2 followed by 250 mg/kg of cyclophosphamide i.p. on day -1. Marrow was prepared from the femurs of donors and infused i.v. into the recipient on day 0. In all, 15 rats treated with Bu-Cy without marrow died, while 22 of 25 transplanted rats survived. In total, 16 rats with measurable tumors showed tumor responses following transplantation, but tumors recurred and survival was minimally prolonged. Of nine rats transplanted before clinical tumors were detected, five became long-term survivors that resisted further tumor challenge. It was concluded that the DMBA-4 breast cancer in WF rats could serve to evaluate SBMT following myeloablative doses of chemotherapy at various tumor loads. At large tumor loads therapy was not curative, but at low tumor burdens cures were possible and resistance to subsequent tumor challenge was demonstrated. The model may be useful for further studies of stem cell infusion in rodent tumor systems. PMID- 12774058 TI - A randomized phase III clinical trial of autologous blood stem cell transplantation comparing cryopreservation using dimethylsulfoxide vs dimethylsulfoxide with hydroxyethylstarch. AB - Hematopoietic stem cells intended for autologous transplantation are usually cryopreserved in solutions containing 10% dimethylsulfoxide (DMSO, v/v) or 5% DMSO in combination with 6% hydroxyethylstarch (HES, w/v). We performed a single blinded, randomized study comparing these cryoprotectant solutions for patients undergoing autologous peripheral blood stem cell (PBSC) transplantation. A total of 294 patients were evaluable; 148 received cells frozen with 10% DMSO and 146 received cells frozen in 5% DMSO/6% HES. Patients who received cells frozen with the combination cryoprotectant recovered their white blood cell count >or=1.0 x 10(9)/l at a median of 10 days, one day faster than those who received PBSC frozen with DMSO alone (P=0.04). Time to achieve neutrophil counts of >or=0.5 x 10(9) and >or=1.0 x 10(9)/l were similarly faster for the recipients of the cells frozen in the combination solution. This effect was more pronounced for patients who received quantities of CD34+ cells higher than the median for the population. Median time to discontinuation of antibiotic use was also one day faster for the recipients of cells cryopreserved with DMSO/HES (P=0.04). In contrast, median times to recovery of platelet count >or=20 x 10(9)/l were equivalent for each group (10 days; P=0.99) and the median numbers of red cell and platelet transfusions did not differ. PMID- 12774060 TI - Successful second allogeneic peripheral blood stem cell transplantation and donor lymphocyte infusion in patients with relapsed acute leukemia using the same donors as for the initial allogeneic bone marrow transplantation. AB - We report the successful treatment of two acute lympho- blastic leukemia (ALL) patients who relapsed following allogeneic bone marrow transplantation (allo-BMT) with allogeneic peripheral blood sem cell transplantation(allo-PBSCT) and donor lymphocyte infusion (DLI) from the same HLA-identical related donors as those used for the first allo-BMT. The patients relapsed on days 154 and 351 from the initial allo-BMT, respectively. Since conventional reinduction chemotherapy failed, allo-PBSCT was undertaken while the patients were still myelosuppressed immediately after reinduction chemotherapy. To induce and/or enhance GVL effects following allo-PBSCT, we performed rapid tapering of CsA and added DLI. After allo-PBSCT and DLI, the patients maintained their complete remission at 55 and 48 months post allo-PBSCT, respectively. From these findings, allo-PBSCT and DLI may be a useful treatment strategy for acute leukemia relapsing after allo-BMT. PMID- 12774059 TI - Nonmyeloablative allogeneic hematopoietic stem cell transplantation for congenital sideroblastic anemia. AB - Congenital sideroblastic anemia (CSA) is a dyserythropoietic disorder that leads to transfusion dependency and subsequent iron overload. Nonmyeloablative allogeneic hematopoietic stem cell transplantation (NST) was performed for a patient with CSA, who had contraindications to conventional allografting. Conditioning was fludarabine, low-dose total body irradiation and antithymocyte globulin, followed by peripheral blood stem cell transplant. Cyclosporine and mycophenolate mofetil were used for graft-versus-host disease prophylaxis. Complete donor chimerism was observed day +131. Early after transplant, the patient became transfusion independent, allowing a regular phlebotomy program. On day +190, refractory lactic acidosis followed by fatal cardiovascular collapse developed, without evidence of infection. Data from this case demonstrates that NST may correct the erythropoietic defect of CSA. PMID- 12774061 TI - Successful T-cell-replete peripheral blood stem cell transplantation from HLA haploidentical microchimeric mother to daughter with refractory acute lymphoblastic leukemia using reduced-intensity conditioning. AB - A 16-year-old girl with refractory acute lymphoblastic leukemia underwent reduced intensity hematopoietic stem cell transplantation from her two-locus-mismatched haploidentical mother, who was microchimeric for the patient's hematopoietic cells. The conditioning regimen comprised melphalan, fludarabine, and low-dose total body irradiation. Non-T-cell-depleted peripheral blood stem cells were infused with graft-versus-host disease (GVHD) prophylaxis consisting of tacrolimus, prednisolone, and short-course methotrexate. Complete donor-type engraftment without evidence of residual leukemia was confirmed on day 22. Severe GVHD was not observed despite rapid cessation of immunosuppression. The patient remains well in continuous remission 15 months after transplant. This successful experience suggests that maternal hematopoietic stem cell transplants for children, in the presence of microchimerism, may be associated with hyporesponsiveness to the inherited paternal HLA antigens (IPA); preventing severe GVHD. PMID- 12774062 TI - Thalidomide as salvage therapy for VAD-refractory multiple myeloma prior to autologous PBSCT. PMID- 12774064 TI - Mean fluorescence intensity (MFI) of beta2 integrin (CD11a and CD11b) expression on peripheral blood (PB) CD34-positive (CD34(+)) cells in steady state correlates inversely with the total amount of harvested CD34(+) cells. PMID- 12774065 TI - Late-onset Epstein-Barr virus (EBV)-negative extranodal B-precursor lymphoblastic lymphoma of donor origin after hematopoietic stem cell transplantation (HSCT). PMID- 12774066 TI - Backward investment strategy? PMID- 12774067 TI - Twenty years of HIV-1 research: what the future holds. PMID- 12774069 TI - TH1-TH2: a procrustean paradigm. PMID- 12774070 TI - Matching SOCS with function. PMID- 12774071 TI - The choices of a natural killer. PMID- 12774072 TI - Manipulating hematopoietic stem cell amplification with Wnt. PMID- 12774073 TI - Bcl11: sibling rivalry in lymphoid development. PMID- 12774076 TI - Pursuing diversified universities. PMID- 12774075 TI - Understanding the function of CD1-restricted T cells. AB - CD1 molecules bind foreign lipid antigens as they survey the endosomal compartments of infected antigen-presenting cells. Unlike T cells that recognize CD1-restricted foreign lipids, CD1-restricted T cells that are self-antigen reactive function as 'auto-effectors' that are rapidly stimulated to carry out helper and effector functions upon interaction with CD1-expressing antigen presenting cells. The functional distinctions between subsets of CD1-restricted T cells, and the pathways by which these cells both influence the inflammatory and tolerogenic effects of dendritic cells and activate natural killer cells and other lymphocytes, provide insight into how CD1-restricted T cells regulate antimicrobial responses, antitumor immunity and the balance between tolerance and autoimmunity. PMID- 12774077 TI - Recovering from cultural devastation. PMID- 12774078 TI - Virus detectives seek source of SARS in China's wild animals. PMID- 12774079 TI - BSE case rattles Canadian officials. PMID- 12774080 TI - Iraqi science faces lonely road to recovery. PMID- 12774081 TI - Anthropologists cast doubt on human DNA evidence. PMID- 12774082 TI - Deep-sea sub aims to get to the bottom of a muddy issue. PMID- 12774083 TI - Experts blast US decision to back nuclear bunker-busters. PMID- 12774084 TI - NASA aims high with orbital transport system. PMID- 12774085 TI - Legal row looms for gene-map firm. PMID- 12774087 TI - Threat of closure hangs heavy over primate centre. PMID- 12774086 TI - Biologists moot daunting plan to track viruses. PMID- 12774089 TI - Special section on Martian Exploration: Mars attracts! PMID- 12774090 TI - Special section on Martian Exploration: Fantastic journeys. PMID- 12774091 TI - Special section on Martian Exploration: Are you on board? PMID- 12774092 TI - Special section on Martian Exploration: The comeback kids. PMID- 12774093 TI - Challenging the tyranny of impact factors. PMID- 12774094 TI - Editors are meant to be judges, not postmen. PMID- 12774095 TI - The system rewards a dishonest approach. PMID- 12774096 TI - Impact factors aren't top journals' sole attraction. PMID- 12774097 TI - Disruption to science in developing countries. PMID- 12774098 TI - Separate achievements of the Humboldt brothers. PMID- 12774103 TI - In search of clarity. PMID- 12774104 TI - Gene expression: Oestrogen receptor hijacked. PMID- 12774105 TI - Chemistry: Tips for moving single molecules. PMID- 12774106 TI - Evolutionary biology: Fractious phylogenies. PMID- 12774109 TI - Astronomy: Sprinkler system. PMID- 12774107 TI - Earth science: A slice of history. PMID- 12774110 TI - Cell division: Genome maintenance. PMID- 12774112 TI - Forensic psychology: Violence viewed by psychopathic murderers. PMID- 12774113 TI - Prion diseases: BSE in sheep bred for resistance to infection. PMID- 12774114 TI - Mantle thermal pulses below the Mid-Atlantic Ridge and temporal variations in the formation of oceanic lithosphere. AB - A 20-Myr record of creation of oceanic lithosphere is exposed along a segment of the central Mid-Atlantic Ridge on an uplifted sliver of lithosphere. The degree of melting of the mantle that is upwelling below the ridge, estimated from the chemistry of the exposed mantle rocks, as well as crustal thickness inferred from gravity measurements, show oscillations of approximately 3-4 Myr superimposed on a longer-term steady increase with time. The time lag between oscillations of mantle melting and crustal thickness indicates that the mantle is upwelling at an average rate of approximately 25 mm x yr(-1), but this appears to vary through time. Slow-spreading lithosphere seems to form through dynamic pulses of mantle upwelling and melting, leading not only to along-axis segmentation but also to across-axis structural variability. Also, the central Mid-Atlantic Ridge appears to have become steadily hotter over the past 20 Myr, possibly owing to north south mantle flow. PMID- 12774115 TI - Structure of the replicative helicase of the oncoprotein SV40 large tumour antigen. AB - The oncoprotein large tumour antigen (LTag) is encoded by the DNA tumour virus simian virus 40. LTag transforms cells and induces tumours in animals by altering the functions of tumour suppressors (including pRB and p53) and other key cellular proteins. LTag is also a molecular machine that distorts/melts the replication origin of the viral genome and unwinds duplex DNA. LTag therefore seems to be a functional homologue of the eukaryotic minichromosome maintenance (MCM) complex. Here we present the X-ray structure of a hexameric LTag with DNA helicase activity. The structure identifies the p53-binding surface and reveals the structural basis of hexamerization. The hexamer contains a long, positively charged channel with an unusually large central chamber that binds both single stranded and double-stranded DNA. The hexamer organizes into two tiers that can potentially rotate relative to each other through connecting alpha-helices to expand/constrict the channel, producing an 'iris' effect that could be used for distorting or melting the origin and unwinding DNA at the replication fork. PMID- 12774116 TI - A class of compact dwarf galaxies from disruptive processes in galaxy clusters. AB - Dwarf galaxies have attracted increased attention in recent years, because of their susceptibility to galaxy transformation processes within rich galaxy clusters. Direct evidence for these processes, however, has been difficult to obtain, with a small number of diffuse light trails and intra-cluster stars being the only signs of galaxy disruption. Furthermore, our current knowledge of dwarf galaxy populations may be very incomplete, because traditional galaxy surveys are insensitive to extremely diffuse or compact galaxies. Aware of these concerns, we recently undertook an all-object survey of the Fornax galaxy cluster. This revealed a new population of compact members, overlooked in previous conventional surveys. Here we demonstrate that these 'ultra-compact' dwarf galaxies are structurally and dynamically distinct from both globular star clusters and known types of dwarf galaxy, and thus represent a new class of dwarf galaxy. Our data are consistent with the interpretation that these are the remnant nuclei of disrupted dwarf galaxies, making them an easily observed tracer of galaxy disruption. PMID- 12774117 TI - Antiferromagnetic order as the competing ground state in electron-doped Nd1.85Ce0.15CuO4. AB - Superconductivity in the high-transition-temperature (high-T(c)) copper oxides competes with other possible ground states. The physical explanation for superconductivity can be constrained by determining the nature of the closest competing ground state, and establishing if that state is universal among the high-T(c) materials. Antiferromagnetism has been theoretically predicted to be the competing ground state. A competing ground state is revealed when superconductivity is destroyed by the application of a magnetic field, and antiferromagnetism has been observed in hole-doped materials under the influence of modest fields. None of the previous experiments have revealed the quantum phase transition from the superconducting state to the antiferromagnetic state, because they failed to reach the upper critical field B(c2). Here we report the results of transport and neutron-scattering experiments on electron-doped Nd1.85Ce0.15CuO4 (refs 13, 14), where B(c2) can be reached. The applied field reveals a static, commensurate, anomalously conducting long-range ordered antiferromagnetic state, in which the induced moment scales approximately linearly with the field strength until it saturates at B(c2). This and previous experiments on the hole-doped materials therefore establishes antiferromagnetic order as a competing ground state in the high-T(c) copper oxide materials, irrespective of electron or hole doping. PMID- 12774118 TI - Selectivity in vibrationally mediated single-molecule chemistry. AB - The selective excitation of molecular vibrations provides a means to directly influence the speed and outcome of chemical reactions. Such mode-selective chemistry has traditionally used laser pulses to prepare reactants in specific vibrational states to enhance reactivity or modify the distribution of product species. Inelastic tunnelling electrons may also excite molecular vibrations and have been used to that effect on adsorbed molecules, to cleave individual chemical bonds and induce molecular motion or dissociation. Here we demonstrate that inelastic tunnelling electrons can be tuned to induce selectively either the translation or desorption of individual ammonia molecules on a Cu(100) surface. We are able to select a particular reaction pathway by adjusting the electronic tunnelling current and energy during the reaction induction such that we activate either the stretching vibration of ammonia or the inversion of its pyramidal structure. Our results illustrate the ability of the scanning tunnelling microscope to probe single-molecule events in the limit of very low yield and very low power irradiation, which should allow the investigation of reaction pathways not readily amenable to study by more conventional approaches. PMID- 12774119 TI - Impact of urbanization and land-use change on climate. AB - The most important anthropogenic influences on climate are the emission of greenhouse gases and changes in land use, such as urbanization and agriculture. But it has been difficult to separate these two influences because both tend to increase the daily mean surface temperature. The impact of urbanization has been estimated by comparing observations in cities with those in surrounding rural areas, but the results differ significantly depending on whether population data or satellite measurements of night light are used to classify urban and rural areas. Here we use the difference between trends in observed surface temperatures in the continental United States and the corresponding trends in a reconstruction of surface temperatures determined from a reanalysis of global weather over the past 50 years, which is insensitive to surface observations, to estimate the impact of land-use changes on surface warming. Our results suggest that half of the observed decrease in diurnal temperature range is due to urban and other land use changes. Moreover, our estimate of 0.27 degrees C mean surface warming per century due to land-use changes is at least twice as high as previous estimates based on urbanization alone. PMID- 12774120 TI - Field sports and conservation in the United Kingdom. AB - Many natural habitats exist on privately owned land outside protected areas, but few governments can afford to enforce or subsidize conservation of this biodiversity. Even in some developed countries, conservation subsidy schemes have only achieved limited success. Fortunately, some landowners may be willing to accept management costs in return for other benefits, although this remains controversial when it involves the killing of charismatic species. For example, participants in British field sports, such as fox hunting and game-bird shooting, may voluntarily conserve important habitats that are required by quarry species. Here we report results from a multidisciplinary study that addressed this issue by focusing on three sites across central England. We found that landowners participating in field sports maintained the most established woodland and planted more new woodland and hedgerows than those who did not, despite the equal availability of subsidies. Therefore, voluntary habitat management appears to be important for biodiversity conservation in Britain. Current debates on the future of field sports in Britain, and similar activities globally, may benefit from considering their utility as incentives to conserve additional habitat on private land. PMID- 12774121 TI - Action video game modifies visual selective attention. AB - As video-game playing has become a ubiquitous activity in today's society, it is worth considering its potential consequences on perceptual and motor skills. It is well known that exposing an organism to an altered visual environment often results in modification of the visual system of the organism. The field of perceptual learning provides many examples of training-induced increases in performance. But perceptual learning, when it occurs, tends to be specific to the trained task; that is, generalization to new tasks is rarely found. Here we show, by contrast, that action-video-game playing is capable of altering a range of visual skills. Four experiments establish changes in different aspects of visual attention in habitual video-game players as compared with non-video-game players. In a fifth experiment, non-players trained on an action video game show marked improvement from their pre-training abilities, thereby establishing the role of playing in this effect. PMID- 12774122 TI - Mitochondrial membrane remodelling regulated by a conserved rhomboid protease. AB - Rhomboid proteins are intramembrane serine proteases that activate epidermal growth factor receptor (EGFR) signalling in Drosophila. Rhomboids are conserved throughout evolution, and even in eukaryotes their existence in species with no EGFRs implies that they must have additional roles. Here we report that Saccharomyces cerevisiae has two rhomboids, which we have named Rbd1p and Rbd2p. RBD1 deletion results in a respiratory defect; consistent with this, Rbd1p is localized in the inner mitochondrial membrane and mutant cells have disrupted mitochondria. We have identified two substrates of Rbd1p: cytochrome c peroxidase (Ccp1p); and a dynamin-like GTPase (Mgm1p), which is involved in mitochondrial membrane fusion. Rbd1p mutants are indistinguishable from Mgm1p mutants, indicating that Mgm1p is a key substrate of Rbd1p and explaining the rbd1Delta mitochondrial phenotype. Our data indicate that mitochondrial membrane remodelling is regulated by cleavage of Mgm1p and show that intramembrane proteolysis by rhomboids controls cellular processes other than signalling. In addition, mitochondrial rhomboids are conserved throughout eukaryotes and the mammalian homologue, PARL, rescues the yeast mutant, suggesting that these proteins represent a functionally conserved subclass of rhomboid proteases. PMID- 12774123 TI - Role of ERas in promoting tumour-like properties in mouse embryonic stem cells. AB - Embryonic stem (ES) cells are pluripotent cells derived from early mammalian embryos. Their immortality and rapid growth make them attractive sources for stem cell therapies; however, they produce tumours (teratomas) when transplanted, which could preclude their therapeutic usage. Why ES cells, which lack chromosomal abnormalities, possess tumour-like properties is largely unknown. Here we show that mouse ES cells specifically express a Ras-like gene, which we have named ERas. We show that human HRasp, which is a recognized pseudogene, does not contain reported base substitutions and instead encodes the human orthologue of ERas. This protein contains amino-acid residues identical to those present in active mutants of Ras and causes oncogenic transformation in NIH 3T3 cells. ERas interacts with phosphatidylinositol-3-OH kinase but not with Raf. ERas-null ES cells maintain pluripotency but show significantly reduced growth and tumorigenicity, which are rescued by expression of ERas complementary DNA or by activated phosphatidylinositol-3-OH kinase. We conclude that the transforming oncogene ERas is important in the tumour-like growth properties of ES cells. PMID- 12774124 TI - Modulation of oestrogen receptor signalling by association with the activated dioxin receptor. AB - Environmental contaminants affect a wide variety of biological events in many species. Dioxins are typical environmental contaminants that exert adverse oestrogen-related effects. Although their anti-oestrogenic actions are well described, dioxins can also induce endometriosis and oestrogen-dependent tumours, implying possible oestrogenic effects. However, the molecular mechanism underlying oestrogen-related actions of dioxins remains largely unknown. A heterodimer of the dioxin receptor (AhR) and Arnt, which are basic helix-loop helix/PAS-family transcription factors, mediates most of the toxic effects of dioxins. Here we show that the agonist-activated AhR/Arnt heterodimer directly associates with oestrogen receptors ER-alpha and ER-beta. This association results in the recruitment of unliganded ER and the co-activator p300 to oestrogen-responsive gene promoters, leading to activation of transcription and oestrogenic effects. The function of liganded ER is attenuated. Oestrogenic actions of AhR agonists were detected in wild-type ovariectomized mouse uteri, but were absent in AhR-/- or ER-alpha-/- ovariectomized mice. Our findings suggest a novel mechanism by which ER-mediated oestrogen signalling is modulated by a co-regulatory-like function of activated AhR/Arnt, giving rise to adverse oestrogen-related actions of dioxin-type environmental contaminants. PMID- 12774127 TI - Thinking ahead. PMID- 12774125 TI - Structure and function of Nurr1 identifies a class of ligand-independent nuclear receptors. AB - Members of the nuclear receptor (NR) superfamily of transcription factors modulate gene transcription in response to small lipophilic molecules. Transcriptional activity is regulated by ligands binding to the carboxy-terminal ligand-binding domains (LBDs) of cognate NRs. A subgroup of NRs referred to as 'orphan receptors' lack identified ligands, however, raising issues about the function of their LBDs. Here we report the crystal structure of the LBD of the orphan receptor Nurr1 at 2.2 A resolution. The Nurr1 LBD adopts a canonical protein fold resembling that of agonist-bound, transcriptionally active LBDs in NRs, but the structure has two distinctive features. First, the Nurr1 LBD contains no cavity as a result of the tight packing of side chains from several bulky hydrophobic residues in the region normally occupied by ligands. Second, Nurr1 lacks a 'classical' binding site for coactivators. Despite these differences, the Nurr1 LBD can be regulated in mammalian cells. Notably, transcriptional activity is correlated with the Nurr1 LBD adopting a more stable conformation. Our findings highlight a unique structural class of NRs and define a model for ligand-independent NR function. PMID- 12774128 TI - Short supply. PMID- 12774129 TI - Growing pains. PMID- 12774130 TI - Prevention of early-onset GBS sepsis: evaluation of a changing paradigm. PMID- 12774131 TI - Neonatal early-onset group B streptococcal disease in the era of intrapartum chemoprophylaxis: residual problems. AB - OBJECTIVE: To identify limitations of current strategies for intrapartum prophylaxis of neonatal early-onset group B streptococcal infection. METHODS: Retrospective review of infants with culture-proven early-onset group B streptococcal infection admitted to two nurseries and their mothers from July 1992, when ACOG and AAP guidelines for intrapartum prophylaxis were first issued, through December 2001. Information was recorded regarding clinical risk factors for early-onset group B streptococcal infection, collection and processing of specimens to assess maternal colonization, delivery of prophylaxis, duration of hospitalization before delivery, and outcome. RESULTS: Among 92 infants with early-onset group B streptococcal infection admitted from 23 institutions, 68 had received no intrapartum prophylaxis. Of these 68 who received no prophylaxis, 34 had identifiable risk factors before delivery (32 clinical, two positive maternal culture), while 34 had no risk factors. Prenatal culture for group B streptococcal colonization was performed in 22 of these women. Of the 18 cultures that were negative for group B streptococcus, 15 were obtained using suboptimal culture technique or were collected more than 6 weeks before delivery. Of the 68 with no prophylaxis, 14 required extracorporeal membrane oxygenation and three died. Of the 24 who received some intrapartum prophylaxis, nine had received >/=two doses for >/=4 hours immediately before delivery. Among the 24 receiving some intrapartum prophylaxis, two required extracorporeal membrane oxygenation and one died. No deaths occurred in those who received >4 hours of prophylaxis, although one such infant required extracorporeal membrane oxygenation. After the CDC guidelines were issued in May 1996, there was a decrease both in the number of cases of early-onset group B streptococcal infection (56 versus 36) as well as in the number with clinical risk factors but no intrapartum prophylaxis (24/56 (43%) versus 5/28 (18%)). CONCLUSIONS: The use of clinical risk factors alone will inevitably result in missed opportunity for intrapartum antibiotic prophylaxis. With maternal screening, false-negative results will be reduced but not necessarily eliminated by assuring that specimens are obtained from proper sites using selective media within 6 weeks of delivery. Better strategies are needed to assure timely administration when prophylaxis is indicated. The nine neonates with early-onset group B streptococcal infection despite intrapartum antibiotics for the recommended duration illustrate that disease may occur even when guidelines are implemented appropriately. PMID- 12774132 TI - "Real World" compliance with strategies to prevent early-onset group B streptococcal disease. AB - OBJECTIVE: To assess the "real-world" compliance with risk- and culture-based strategies to prevent early-onset group B streptococcal disease. STUDY DESIGN: We retrospectively reviewed the medical records of consecutive term pregnancies delivered at three institutions (a subset of practices at an academic hospital using the culture-based strategy, an academic hospital using the risk-based strategy, and a community hospital using both) between January and March 1998. We abstracted demographic data and risk factors for group B streptococcus, group B streptococcal culture information, documentation of intrapartum antibiotic prophylaxis, and adverse drug reactions. We compared intrapartum compliance with the intended strategy. RESULTS: There were a total of 505 women managed with the risk-based strategy. Of those, 79 had a risk factor for group B streptococcal disease and 72/79 (91.1%) received intrapartum antibiotic prophylaxis. There were a total of 428 women managed with the culture-based strategy. Of those, 70 had positive cultures and 67 (95.7%) received intrapartum antibiotic prophylaxis. An additional 39 women in the culture-based group had no documentation that cultures had been done. Of those, eight (20.5%) had risk factors and all eight received intrapartum antibiotic prophylaxis. Compliance with the risk-based strategy was 91.1 versus 96.2% with the culture-based strategy (p=0.3). Among women managed using the risk-based strategy, 5/426 (1.2%) received intrapartum antibiotic prophylaxis without an identifiable risk factor. Among women in the culture-based strategy, 5/359 (1.4%) received intrapartum antibiotic prophylaxis with documented negative group B streptococcal cultures (p=0.5). When examined by site, compliance with the intended strategy was 91.2% at the academic hospital using the risk-based strategy, 100% at the academic hospital using the culture based strategy, 90.9% at the community practices using the risk-based strategy, and 82.4% at the community practices using the culture-based strategy. CONCLUSION: Overall, intrapartum compliance with the risk-based approach was similar to the culture-based approach. However, there were more cultures not done and cultures done at inappropriate gestations at the community hospital practice. PMID- 12774133 TI - Necrotizing enterocolitis among neonates in the United States. AB - BACKGROUND: Prior studies have identified individual risk factors that are associated with necrotizing enterocolitis (NEC); however, the small sample sizes of these previous studies have not allowed the analysis of potential interaction between multiple variables and NEC. Our purpose was to describe the incidence and risk factors for NEC in premature neonates admitted for intensive care. METHODS: We identified neonates as having NEC if they met accepted diagnostic criterion for necrotizing enterocolitis. Using a national database, we assessed the association between NEC and a battery of risk factors previously reported in peer reviewed literature. RESULTS: There were 15,072 neonates that met inclusion criteria; 14,682 did not have NEC, while 390 (2.6%) met criterion for NEC. Multivariate analysis showed that low birth weight was the most important risk factor for NEC. Other factors that were associated with an increased risk of NEC were exposure to antenatal glucocorticoids, vaginal delivery, need for mechanical ventilator support, exposure to both glucocorticoids and indomethacin during the first week of life, absence of an umbilical arterial catheter, and low Apgar score at 5 minutes. Length of hospital stay and mortality were higher in neonates with NEC than in neonates without NEC. CONCLUSIONS: NEC remains an important cause of morbidity and mortality in prematurely born neonates. In contrast to previous studies, we found that exposure to antenatal glucocorticoids was associated with an increased risk for NEC independent of birth weight. PMID- 12774134 TI - Necrotizing enterocolitis and gastrointestinal complications after indomethacin therapy and surgical ligation in premature infants with patent ductus arteriosus. AB - BACKGROUND: Indomethacin is the most frequently used pharmacological agent for closure of a patent ductus arteriosus (PDA) in premature infants. However, reports of complications, particularly, necrotizing enterocolitis (NEC) and isolated gastrointestinal perforation have generated concerns about the use of this medication. OBJECTIVES: A retrospective study to compare the incidence of NEC, NEC-related gastrointestinal complications and isolated gastrointestinal perforation among premature infants treated for a PDA with either, indomethacin alone (I), surgical ligation alone (L), or indomethacin followed by surgical ligation (I-L). METHODS: The medical records of 224 infants that underwent treatment, either pharmacological or surgical, for a PDA, confirmed by echocardiography, over a 4-year period (1995 to 1998) were analyzed. Treatment history and gastrointestinal complications were reviewed. RESULTS: Of the 224 infants, 108 (48.2%) were treated with I, 50 (22.3%) by L, 66 (29.5%) with I-L. The clinical characteristics of the three treatment groups were similar and no differences in the incidence of NEC were observed between groups. NEC occurred in 14 (13%) of the I group, seven (14%) of the L group, and eight (12%) of the I-L group. The rate of NEC related gastrointestinal complications and isolated gastrointestinal perforation were also similar among groups. CONCLUSION: In this large retrospective study, indomethacin treatment for a significant PDA in premature infants was not associated with a greater risk for NEC or NEC-related gastrointestinal complications than surgical ligation. PMID- 12774135 TI - Rising birth prevalence of gastroschisis. AB - OBJECTIVE: Gastroschisis is a congenital anomaly that has been reported to be increasing in frequency. The objective of this study was to determine the birth prevalence of gastroschisis using two large databases. STUDY DESIGN: We reviewed data from a statewide database and a national database from a neonatal health care provider, abstracting cases of gastroschisis. RESULTS: In North Carolina, the birth prevalence of gastroschisis increased from 1.96 per 10,000 births in 1997 to 4.49 per 10,000 births in 2000 (p=0.0007). The overall increase was almost entirely because of the increase in infants born to mothers less than 20 years old. Among infants receiving care from the national neonatal provider, the prevalence of gastroschisis increased from 2.9 per 1000 patients in 1997 to five per 1000 patients in 2001 (p=0.044). CONCLUSION: The birth prevalence of gastroschisis is increasing in North Carolina, and this trend may be occurring nationally. The rapid change in the birth prevalence in the subset of population most at risk for gastroschisis implicates environmental or pharmacologic teratogens rather than changing population characteristics as a causal factor in the development of gastroschisis. PMID- 12774136 TI - Age-related low expression of HLA-DR molecules on monocytes of term and preterm newborns with and without signs of infection. AB - OBJECTIVES: Presentation of human leucocyte antigen (HLA) molecules is an important part of an efficient immune response. Since bacterial infections are more common in newborns, we hypothesized that the level of HLA-DR expression may influence the host defense system. STUDY DESIGN: HLA-DR expression on monocytes was examined by flow cytometry during the first week of life of term and preterm neonates with and without signs of infection and of adults. RESULTS: HLA-DR expression of term and preterm newborns with or without signs of infection was lower compared with adults during the first day of life (p<0.0001). Prematurity correlates with lower expression in neonates with gestational age less than 32 weeks (p=0.0008). HLA-DR expression in neonates with signs of infection was decreased compared to healthy neonates (p=0.0196). Maternal conditions such as preeclampsia, prenatal treatment with steroids and mode of delivery had no influence on the expression of HLA-DR. In contrast, newborns with respiratory distress syndrome but without signs of infection showed reduced HLA-DR expression (p=0.0370). CONCLUSION: Low HLA-DR expression on monocytes contributes to impaired neonatal host defense, especially in preterm neonates. PMID- 12774137 TI - Low-dose nitric oxide therapy for persistent pulmonary hypertension: 1-year follow-up. AB - BACKGROUND: Although inhaled nitric oxide has been shown to reduce the use of extracorporeal membrane oxygenation, there is limited data on its effect on long term outcomes. OBJECTIVE: The purpose of our study is to report on the 1 year outcome of neonates treated with inhaled nitric oxide compared to a group of neonates who did not receive nitric oxide. METHODS: We have previously reported on the in-hospital outcomes of 248 neonates who were >34 weeks' gestational age and were randomized to receive low-dose inhaled nitric oxide or placebo. Extracorporeal membrane oxygenation was used in 78 (64%) neonates in the control group and in 48 (38%) neonates in the inhaled nitric oxide group (p=0.001). We now report on the outcome of neonates followed to 1 year of age. RESULTS: Of the 248 neonates twenty-four (10%) died before 1 year of age. There was no difference in mortality between the two groups (11% in the control group and 9% in the inhaled nitric oxide group). Of the 224 surviving infants, we were able to contact the parents or guardians of 201 (90%) children. There were no intergroup differences in the numbers of patients reported as requiring medications for pulmonary disease (14% in the control group and 14% in the inhaled nitric oxide group) or the need for supplemental oxygen (1% in the control group and 0% in the inhaled nitric oxide group). The number of neonates reported to have an abnormal neurological examination or developmental delay was also similar in both groups (14% in the control group and 19% in the inhaled nitric oxide group). CONCLUSIONS: The use of low-dose inhaled nitric oxide reduces the use of extracorporeal membrane oxygenation without increasing the incidence of adverse outcomes to 1 year of age. PMID- 12774139 TI - Brainstem maturation after antenatal steroids exposure in premature infants as evaluated by auditory brainstem-evoked response. AB - OBJECTIVE: Antenatal steroids result in fetal lung maturation, but may retard brain development. Auditory brainstem-evoked response (ABR) is a noninvasive assessment of brainstem maturation. The objective of this study was to determine if antenatal steroids affect brainstem maturation in infants 0.022 for LW/BW and from 2.2 to 4.4 for RAC. RESULTS: In 17 cases (65%) pulmonary hypoplasia was observed, characterized by a LW/BW value below 0.022 and RAC below 2.2. In nine cases (35%), microscopic examination of brainstem serial sections showed varying degrees of hypoplasia of the arcuate nucleus (ARCn). In eight cases (31%) the pulmonary hypoplasia was associated with hypoplasia/agenesis of the ARCn. CONCLUSIONS: This study demonstrated that in about a third of stillbirths there is a congenital hypodevelopment of both lung and arcuate nucleus. In these cases the ARCn hypoplasia would exert a negative effect on respiratory movements in utero and therefore on lung development. When the pulmonary hypoplasia is not accompanied by hypodevelopment of this nucleus the explanation could be a failure to block the inhibitory action of the Kolliker-Fuse nucleus. PMID- 12774144 TI - Detoxifying death in the neonate: in search of meaningfulness at the end of life. AB - At the end of life, when curative measures have failed to conquer the terminal nature of a disease, it is imperative to introduce healing measures that deal with the human suffering associated with illness. When dealing with critically ill neonates with lethal conditions, it may be necessary to institute healing measures at the beginning of life. In such cases, healing measures must incorporate customary palliative measures but should not be limited to them. The purpose of this paper is to describe two families' approaches to palliation at the end of life in their newborn infants with Trisomy 18, a genetic abnormality usually associated with a markedly decreased life expectancy. We shall consider the concepts of interference versus intervention as we examine subtle medical differences between the two cases. We will address medical, legal, and ethical issues in each case, but special attention will be given to the provision of compassionate care. People face physical, mental, emotional, and spiritual challenges as they go through life. The families' approaches with their infants at the end of their lives may support the premise that the latter two challenges, emotional and spiritual, confront us the most at the end. Encouraging families to engage with their dying infants helps detoxify the experience and make it more meaningful. PMID- 12774145 TI - Nutrition in the neonatal intensive care unit: how do we reduce the incidence of extrauterine growth restriction? AB - Extrauterine growth restriction is a major clinical problem for prematurely born neonates, especially critically ill preterm neonates, and malnutrition in the neonatal intensive-care unit remains common. There are numerous perceived risks to initiation of adequate nutritional support. How many of these factors pose a real risk to health outcomes is less clear. Current nutritional support does not prevent extrauterine growth restriction and the consequences of malnutrition are both acute and delayed. Our clinical approach to providing nutritional support impacts neonatal morbidity and long-term neuro developmental outcomes. While more and better evidence is needed to help guide best practices, this gap should not prevent neonatologists from using the observations in this review to improve their current practice. There is evidence that changes in nutritional support can have a positive influence on growth. These include early administration of intravenous amino acids and lipids, minimal enteral nutrition, and supplemented formula and human milk. Simply recognizing the degree of growth failure by monitoring weight and focusing on the accruing deficit should encourage clinicians to increase nutritional support to enhance recovery growth. Continued research is needed to define the efficiency of early feeding, more rapid advancements in nutritional support, protein needs, the optimal composition of breast-milk supplements, the etiology of necrotizing enterocolitis, and perhaps most importantly, the health consequences of extrauterine growth restriction. PMID- 12774146 TI - Neonatal gastric perforation. PMID- 12774147 TI - Resolution of peripheral artery catheter-induced ischemic injury following prolonged treatment with topical nitroglycerin ointment in a newborn: a case report. AB - Tissue ischemia, necrosis, and gangrene are uncommon but well-described complications of arterial catheterization in the neonate. Treatment options for progressive tissue necrosis following arterial embolization and/or vasospasm are limited in these patients secondary to unpredictable pharmacokinetics and risks associated with systemic anticoagulation or vasodilatation in newborns. We report a case of a multidose regimen of topical 2% nitroglycerin ointment for reversing severe tissue ischemia following peripheral arterial line placement. The favorable response in this infant suggests that topical nitroglycerin therapy should be considered as potential therapy to ameliorate the effects of vascular compromise following arterial line placement in neonates. PMID- 12774148 TI - Vernixuria: another sign of uterine rupture. AB - Uterine rupture complicates approximately 1% of trials of labor after cesarean. Classic signs and symptoms include loss of station, cessation of labor, vaginal bleeding, fetal distress, and abdominal pain. Other signs are also possible. We report a case of uterine rupture at VBAC trial that includes an unusual clinical sign of uterine rupture: vernix caseosa observed in the urine of the parturient. During labor, a bladder catheter was inserted to evaluate oliguria. Vernix caseosa and blood were found in the tubing. Prompt cesarean delivery followed. A tear extending from the original transverse scar into the bladder dome was found. Vernixuria is an additional sign of uterine rupture. PMID- 12774149 TI - Improvements in surgical technique of valgus high tibial osteotomy. AB - We present four technical modifications of high tibial osteotomy which improve its safety and reproducibility. (a) Open wedge correction: opening wedge osteotomy from the medial side avoids lateral muscle detachment, dissection of the peroneal nerve, proximal fibula osteotomy, and leg shortening; only one osteotomy needs to be performed and the correction can be adapted intraoperatively. (b) Biplanar osteotomy: in addition to the transverse osteotomy of the posterior tibia a second ascending osteotomy in the coronary plane underneath the tibial tuberosity is performed. This provides improved rotational stability of the osteotomy and creates an anterior buttress against sagittal tilting of the osteotomy planes. (c) Incomplete osteotomy with plastic deformation of the tibia: 10 mm of lateral bone stock is left intact. The osteotomy is opened gradually over several minutes by sequential impaction of flat chisels or by use of a special spreading tool. Manifest fractures of the lateral cortex with resulting instability are avoided. Rapid bone healing is promoted. (d) Rigid fixation: stable osteosynthesis allows for early mobilization and avoids losses-of-correction. We use a medial plate-fixator which can be applied percutanously. In 112 patients operated on using this modified technique no pseudarthosis or loss-of-correction was observed. PMID- 12774150 TI - An innovative compression-clamp for open contaminated fractures of patella: a short report of five cases. AB - Five patients with open contaminated fractures of the patella were managed using an indigenously designed external clamp cum compressor. All had an infected lacerated wound at the fracture site, and one had open exposed knee joint (Gustilo grade IIIB open fracture). These patients reported after 3-7 days of injury. The wounds were thoroughly washed and debrided and the clamp cum compressor was applied. They were encouraged to do active knee movements immediately post-operatively. The wounds healed within 3-4 weeks, and the fractures united within 10-16 weeks. Good functional results were achieved in four of the five. PMID- 12774151 TI - Blood loss in patients for total knee arthroplasty. AB - Ninety-four patients with osteoarthritis (OA) and 180 with rheumatoid arthritis (RA) undergoing unilateral total knee arthroplasty (TKA) were analyzed to clarify the necessity for preoperative autogenous blood deposition or homologous blood transfusion. Two hundred and twenty-four and 50 patients underwent TKA with cement and without cement, respectively. The difference in average blood loss in patients between with (372 ml) and without cementation (449 ml) was significant. In the OA group the average blood loss significantly decreased after cementation but not in the RA group. Although the rate of avoiding transfusion in the OA group did not significantly decrease with the use of cement (92.4% vs. 93.3%), that in the RA group did (80% to 57.1%). Eight of 159 patients with hemoglobin level (Hb) of 11.0 g/dl or higher received homologous blood transfusion. Of these eight patients five had associated disorders. Only one patient with Hb of 12.0 g/dl or higher underwent homologous blood transfusion. Patients with Hb of 12.0 g/dl or higher are not indicated for preoperative autologous blood deposition. In patients with Hb between 11.0 and 12.0 g/dl preoperative blood deposition may be planned after consideration of general condition and complication. Patients with Hb lower than 11.0 g/dl should undergo preoperative blood deposition. PMID- 12774152 TI - A biomechanical and histological evaluation of the structure and function of the healing medial collateral ligament in a goat model. AB - This study evaluated the healing process of an isolated medial collateral ligament (MCL) rupture at 12 weeks in a goat model. Using a robotic/UFS testing system, knee kinematics in multiple degrees of freedom and in situ forces in the healing MCL in response to (1) a 67-N anterior tibial load and (2) a 5-Nm valgus moment were evaluated as a function of angles of knee flexion. Then a uniaxial tensile test of femur-MCL-tibia complexes (FMTCs) was preformed to obtain the structural properties of the FMTC and mechanical properties of the healing MCL substance. The histological appearance of the healing MCL was also examined for collagen and cell organization. The anterior tibial translation in response to a 67-N anterior tibial load was found to range from 1.9 to 2.4 mm, which was not significantly different from the sham-operated, contralateral control knee. In response to a 5-Nm valgus moment, however, MCL injury caused a 40% or more increase in valgus rotations over sham-operated controls for all angles of knee flexion tested. The magnitudes of the in situ forces in the healing MCLs for neither external loading conditions differed from sham-operated controls. For the structural properties of the healing FMTC, the stiffness returned to sham operated control levels, but ultimate load at failure remained 60% of sham operated control values. In terms of mechanical properties of the healing MCL, its tangent modulus and stress at failure were only 40% of sham-operated control values. Histologically, the collagen and cell organization at the femoral and tibial insertions as well as the midsubstance remained disorganized. Comparing these data to those previously reported at 6 weeks, there was a marked improvement in the in situ forces in the healing MCL and of the stiffness of the FMTC. Also, the data obtained for the goat model revealed a faster healing process than those for the rabbit model. These findings suggest that greater post injury activity levels may render the goat to be a better animal model for studying the healing process of the MCL. PMID- 12774153 TI - Simultaneous bilateral tibial tubercle avulsion fracture in a basketball player. AB - A 16-year-old male basketball player had sustained an injury upon landing after a forceful jump. Plain radiography demonstrated bilateral tibial tubercle avulsion fracture involving partially proximal physis. Open reduction and internal fixation were performed at once. Continuous passive motion was started immediately after operation, and the patient was ambulated with hinged knee extension braces. After 27 months follow-up his knees completely regained normal range of motion except a 3 degrees extension loss in the left knee. He resumed all daily functional activities (Lysholm functional score of 99), but he slightly lost his level of activity (Tegner activity level from 7 to 6). No angular deformity at all on the frontal plane was determined upon radiological examination. Tibial slope angles were symmetrical and within the normal range. There were visible small bone fragments inside the left patellar tendon. PMID- 12774154 TI - Outside-in meniscus suture technique: 5 years' follow-up. AB - We present the results of a prospective study evaluating the arthroscopic technique of outside-in meniscus suture (n=68). The types of tears suitable for suture consisted of acute vertical tears, solitary or in association with a radial tear in middle third. The technique was always outside-in using resorbable sutures (2-0 PDS, 2-0 Maxon). Follow-up evaluation included clinical examination, radiographic examinations and second-look arthroscopies (n=10). There were four failures in this study, consisting of rerupture of the meniscus. Clinical results were good in 91% with complete recovery and return to physical and sportive activity, good in 3% with mild symptoms, and failure in 6% consisting of meniscus rerupture. PMID- 12774155 TI - Is low monocyte HLA-DR expression helpful to predict outcome in severe sepsis? AB - OBJECTIVE: Low monocyte human leukocyte antigen-DR (HLA-DR) expression has been reported to be an indicator of poor survival in critically ill septic patients. We assessed its usefulness as a prognostic indicator in order to identify possible interventions to normalise HLA-DR expression in those patients with lowered monocyte HLA-DR. DESIGN: HLA-DR expression was measured on separated monocytes of septic patients, using flow cytometry, and HLA-DR upregulation was measured by the same techniques after ex vivo stimulation with granulocyte macrophage colony stimulating factor (GM-CSF). APACHE II score, age, sex and outcome were determined for all patients. SETTING: A single-centre study at the Royal Liverpool University Hospital in a medico-surgical 13-bed intensive care unit. PATIENTS AND PARTICIPANTS: All septic patients ( n=70) fulfilling the criteria of the ACCP for the diagnosis of sepsis were recruited for the study with informed consent from day 1 of diagnosis of sepsis and monocyte HLA-DR expression measured on 3 consecutive days. Patients were excluded from the study if they were on immunosuppressive therapy. Normal healthy volunteers ( n=45) were included. RESULTS: Low monocyte surface expression and median fluorescence density HLA-DR expression was not associated with a high mortality. High APACHE II scores were not correlated with low HLA-DR expression. However, in those patients where HLA-DR expression was lowered, this could be restored ex vivo by GM-CSF. CONCLUSIONS: In the group of septic patients under study, HLA-DR was not a useful prognostic marker of outcome. We did not find a higher mortality in the group of patients who had low expression. These findings are contradictory to some previously reported findings, and the possible reasons are discussed. PMID- 12774156 TI - Risk factors for piperacillin/tazobactam-resistant Escherichia coli in ICU patients: a clinical study. AB - OBJECTIVE: To determine risk factors of infections with piperacillin/tazobactam resistant Escherichia coli in critical care patients. DESIGN: Prospective, consecutive sample survey study. SETTING: Surgical intensive care unit (ICU) in a university hospital. PATIENTS: A consecutive series of 133 patients from whom culture results were positive for E. coli during their ICU stay. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Multivariate logistic regression analysis identified the following significant independent factors associated with the emergence of a piperacillin/tazobactam-resistant Escherichia coli: prior use of amoxicillin (odds ratio, 4.15) and amoxicillin/clavulanate (odds ratio, 3.25). CONCLUSIONS: Treatment with amoxicillin or amoxicillin/clavulanate is a major risk factor for the detection of piperacillin/tazobactam-resistant E. coli in ICU patients. PMID- 12774157 TI - The accuracy of transcranial Doppler to detect vasospasm in patients with aneurysmal subarachnoid hemorrhage. AB - OBJECTIVES: To examine the accuracy of transcranial Doppler to detect cerebral vasospasm in a patient population with aneurysmal subarachnoid hemorrhage. DESIGN: Prospective blind comparison of transcranial Doppler with cerebral angiography. Diagnostic accuracy of transcranial Doppler was assessed using receiver operating characteristic (ROC) analysis and likelihood ratios. Sensitivity and specificity were calculated using directly measured middle cerebral artery diameter as reference standard. SETTING: Intensive Care Unit of a large university teaching hospital. PATIENTS AND PARTICIPANTS: Twenty-two patients with subarachnoid hemorrhage were included. Patients underwent angiography on admission and after 8 days to diagnose vasospasm and were defined as having clinical vasospasm, angiographic vasospasm, or no vasospasm. MEASUREMENTS AND RESULTS: Sensitivity and specificity were 1.00 and 0.75 for angiographic vasospasm and both equal to 1.00 for clinical vasospasm diagnosis. A transcranial Doppler mean velocity threshold value of 100 cm/s for angiographic vasospasm and 160 cm/s for clinical vasospasm detection were chosen by ROC analysis. CONCLUSIONS: A Transcranial Doppler mean velocity threshold of 160 cm/s, calculated by the ROC analysis, accurately detects clinical vasospasm. A daily transcranial Doppler examination performed by a trained operator should be routinely used to provide early identification of patients at high risk and to orient therapeutic decisions. PMID- 12774158 TI - Comparison of two different CPAP systems by tidal breathing parameters. AB - OBJECTIVE: Comparison of tidal breathing and pressure fluctuation of the continuous positive airway pressure (CPAP) associated with the use of the valveless Infant Flow System versus the conventional constant-flow CPAP (Babylog 8000) in preterm infants. DESIGN: Randomized cross-over trial. SETTING: Neonatal intensive care unit level III. PATIENTS: Twenty infants; median (range): birth weight 1,035 g (640-4,110 g), actual weight 1,165 g (820-4,250 g), gestational age at birth 27 (26-40) weeks. INTERVENTIONS: After extubation two CPAP devices (Infant Flow System vs Babylog 8000) were applied in a random order to the same infant. Fluctuations of the applied pressure during the breathing cycle and tidal breathing parameters were measured by the flow-through technique. MAIN RESULTS: Using the Infant Flow System the mean (standard deviation) inspiratory flow [1.5 (0.1) vs 1.3 (0.1) l.min(-1).kg(-1), P<0.05] and tidal volume were significantly increased [5.3 (1.3) vs 4.7 (1.3) ml/kg(-1), P<0.05] compared to Babylog 8000. The fluctuations of the applied pressure of the Infant Flow System during the breathing cycle were significantly lower [0.1 (0.03) kPa vs 0.15 (0.08) kPa, P<0.05] compared to Babylog 8000. No differences were seen in the duration of inspiration and expiration and the time to peak tidal flow. In the Infant Flow System pressures during expiration remained stable whereas they increased during the use of Babylog 8000. CONCLUSIONS: Within-subject comparisons of tidal breathing parameters of the two CPAP devices Infant Flow System and Babylog 8000 show: (1) a significant influence of the system used; and (2) that the valveless Infant Flow System increases air flow and tidal volume with less fluctuations in CPAP pressures during the breathing cycle. PMID- 12774159 TI - Randomised trial on the influence of continuous magnesium infusion on arrhythmias following cardiopulmonary bypass surgery for congenital heart disease. AB - OBJECTIVES: To check the hypothesis that continuous magnesium infusion protects the heart from arrhythmias following cardiopulmonary bypass surgery for congenital heart disease. DESIGN: A prospective randomised placebo-controlled study, with patients stratified in three weight groups. PATIENTS AND PARTICIPANTS: The study group ( n=65) postoperatively received a magnesium infusion (1 mmol/kg), the control group ( n=66) received placebo. In both groups serum and ionised magnesium values were followed, and all postoperative arrhythmias were documented for 24 h. MEASUREMENTS AND RESULTS: Serum and ionised magnesium in the blood was elevated after the end of bypass (0.54+/-0.15 mmol l( 1) pre-operatively, 0.88+/-0.24 mmol l(-1) postoperatively), where a cardioplegia solution containing magnesium was used. Magnesium values remained at this elevated level in the magnesium therapy group, and decreased to normal pre operative values within 24 h in controls ( P<0.001). The incidence of postoperative arrhythmias was lower in the study group: 8/65 in the study group and 17/66 in the control group, respectively (chi-squared test, P=0.05). Lower patient weight (32.7 kg versus 22.6 kg), longer cardiopulmonary bypass time (128.7 min versus 87.9 min) and deeper body temperature during extracorporeal circulation (29.2 degrees C versus 32.6 degrees C) were identified as risk factors for postoperative arrhythmias ( P<0.05). CONCLUSIONS: Continuous magnesium infusion effectively reduces the rate of arrhythmias following cardiopulmonary bypass surgery for congenital heart disease and should, therefore, be routinely used. PMID- 12774160 TI - Muscle perfusion and oxygen consumption by near-infrared spectroscopy in septic shock and non-septic-shock patients. AB - OBJECTIVE: To measure muscle blood flow (Qtis) and oxygen consumption (VO(2)tis) in septic and non-septic critically ill patients by near-infrared spectroscopy (NIRS). SETTING: Surgical intensive care unit of a university hospital. PATIENTS AND PARTICIPANTS: Four patients with septic shock, eight post-surgical critically ill patients and ten healthy volunteers. MEASUREMENTS AND RESULTS: Oxyhaemoglobin (HbO(2)) and deoxyhaemoglobin (HbH) variations after venous occlusion were measured by NIRS in the brachioradialis muscle. We calculated Qtis by the rate of HbO(2) and HbH increase in the first 30 s of venous occlusion divided by haemoglobin blood concentration. VO(2)tis was calculated by subtraction of the arterial HbH from the initial increase of HbH after venous occlusion extrapolated to 1 min. Tissue oxygenation index [TOI = HbO(2)/(HbO(2)+HbH)] was also measured before venous occlusion. Two measurements in patients with septic shock, and one measurement in non-septic-shock patients and healthy subjects, were obtained. Of the measurements, 35% were repeated because of low-quality NIRS signal. VO(2)tis and Qtis were two times larger ( P<0.05) in patients with septic shock than in patients without and in healthy subjects. The TOI was very similar among the three groups. CONCLUSION: In septic-shock patients the increase in VO(2)tis was associated with an equivalent increase in Qtis. Therefore, tissue O(2) supply does not seem to be a limiting factor for muscle O(2) consumption. NIRS combined with venous occlusion allows a rapid, non-invasive and simultaneous assessment of regional perfusion and oxygen consumption. In case of microcirculatory shunt occurrence, the TOI should be cautiously used to assess tissue oxygenation state. PMID- 12774161 TI - Quality of life in acute respiratory distress syndrome survivors may be no worst than in other ICU survivors. AB - OBJECTIVE: To compare the health-related quality of life (HR-QOL) in acute respiratory distress syndrome (ARDS) survivors with that in a matched control group of non-ARDS survivors. DESIGN AND SETTING: Prospective, matched, parallel cohort study, comparing HR-QOL between intensive care unit (ICU) survivors with ARDS and a control group in a tertiary care hospital. PATIENTS: Between May 1997 and December 2000, all ARDS adult patients of an eight-bed medical/surgical unit of a tertiary care hospital were enrolled and a control group of non-ARDS survivors, matched for severity of disease and for previous health state, was selected. The study included 29 ARDS survivors who answered the EQ-5D questionnaire and had lung function evaluated. MEASUREMENTS AND RESULTS: A follow up appointment was performed 6 months after ICU discharge consisting of: (a) evaluation of HR-QOL using EQ-5D and (b) lung function tests and measure of diffusing capacity. Among ARDS survivors 41% had normal lung function and 59% mild to moderate lung function impairments. Nearly a one-third of ARDS survivors reported problems in one or more of the five dimensions of the EQ-5D, and 48% reported feeling worse at the interview than 6 month before ICU admission. No significant differences were found in HR-QOL between ARDS survivors and other ICU survivors with similar age and matched for previous health state and severity of disease. CONCLUSIONS: This study suggests that impairments in HR-QOL among ARDS survivors may not be distinguishable from that among other ICU survivors. PMID- 12774162 TI - Plasma oxidative parameters and mortality in patients with severe burn injury. AB - OBJECTIVE: To determine xanthine oxidase and superoxide dismutase activities, lipid peroxidation, protein carbonylation, and total radical-trapping antioxidant parameter in survivors and nonsurvivors patients with severe burn injury. DESIGN AND SETTING: Prospective, comparative observational study in an intensive care unit, burn division, in a trauma hospital. PATIENTS: Twenty-five consecutive patients who met the established criteria for severe burn injury (total burn surface area of more than 30%). MEASUREMENTS AND RESULTS: Plasma thiobarbituric acid reactive species and protein carbonyls levels were significantly higher in nonsurvivors than in survivors at 0 and 6 h. Elevated xanthine oxidase activity at 0 h was associated with adverse outcome after burn injury. In contrast, plasma superoxide dismutase activity and total radical-trapping antioxidant parameter did not differ significantly between nonsurvivors and survivors at any time point. CONCLUSIONS: For the first time we demonstrate the value of oxidative parameters, namely thiobarbituric acid reactive species, protein carbonyls, and xanthine oxidase activity, in identifying burn patients with a poor prognosis. Whether these parameters are merely markers of clinical course, or whether they signal specific deleterious effects of oxidative stress during the burn injury remains to be elucidated. PMID- 12774163 TI - Preventive effect of cerivastatin on diabetic nephropathy through suppression of glomerular macrophage recruitment in a rat model. AB - AIMS/HYPOTHESIS: We investigated the effect of cerivastatin, a statin, on the development of diabetic nephropathy in spontaneously hypertensive rats (SHR) with streptozotocin-induced diabetes. METHODS: Diabetic SHR were given standard chow or chow containing cerivastatin at a dose of 0.1 mg/kg or 1.0 mg/kg for 12 weeks. Effects of cerivastatin on urinary albumin excretion, mesangial expansion, glomerular macrophage infiltration, and the number of anionic sites on the glomerular basement membrane (GBM) were assessed. RESULTS: Cerivastatin did not affect the blood glucose concentration, blood pressure or serum cholesterol concentration in diabetic SHR. However, cerivastatin treatment caused a dose dependent decrease of albuminuria and hyperfiltration. At 1.0 mg/kg, cerivastatin inhibited the diabetes-induced expansion of mesangial and tuft areas on histological examination of the kidneys, as well as the loss of anionic sites from the GBM evaluated with polyetyleneimine and the intraglomerular infiltration of ED1-positive macrophages evaluated by immunohistochemistry. Whole-kidney expression of mRNA for MCP-1 and TGF-beta, estimated by the real-time quantitative RT-PCR, was increased (both 2.6-fold) in untreated diabetic SHR at 12 weeks. Cerivastatin treatment (1.0 mg/kg) inhibited the up-regulated expression of MCP-1 and TGF-beta mRNA (decreased to 48% and 34%, respectively) in diabetic SHR. CONCLUSION/INTERPRETATION: In this hypertensive model of diabetic nephropathy, cerivastatin decreased albuminuria through suppression of glomerular hyperfiltration, mesangial expansion, and the loss of charge barrier independently of a cholesterol-lowering effect. These preventive effects could be at least partly due to inhibition of macrophage recruitment and activation, and inhibition of TGF-beta overexpression. PMID- 12774164 TI - Homocysteine as a risk factor for nephropathy and retinopathy in Type 2 diabetes. AB - AIMS/HYPOTHESIS: The aim of this study was to examine the relation between serum total homocysteine concentrations and microvascular complications in Pima Indians with Type 2 diabetes. METHODS: Homocysteine concentrations were measured in frozen sera of 396 diabetic participants in a longitudinal study who were 40 years of age or older and who had attended one or more examinations between 1982 and 1985. Retinopathy was assessed by fundoscopy and nephropathy by an albumin:creatinine ratio greater than 300 mg/g. The incidence rate ratio for a 5 micro mol/l difference in homocysteine was calculated using proportional hazard regression. RESULTS: The incidence of each complication was assessed in subjects without that complication at baseline and with more than one follow-up examination: 229 for nephropathy, 212 for retinopathy and 266 for proliferative retinopathy. There were 101 incident cases of nephropathy, 113 of retinopathy and 40 of proliferative retinopathy during a mean follow-up of 8.6, 7.5 and 8.9 years, respectively. Incidence of nephropathy was associated with homocysteine concentrations: IRR=1.42 (95% CI, 1.09-1.84, p=0.01); this remained statistically significant controlled for age, sex and duration of diabetes (p=0.03), but not when controlled for baseline renal function (p=0.4). Homocysteine concentrations were not associated with the incidence of any retinopathy IRR=1.14 (95%CI 0.89 1.46, p=0.3) but were associated with the incidence of proliferative retinopathy IRR=1.62 (95% CI 1.16-2.28, p=0.005); this association remained statistically significant when controlled for baseline renal function and diabetes duration (p=0.02). CONCLUSIONS/INTERPRETATION: Increased homocysteine concentrations are associated with an increased risk for incidence of nephropathy and proliferative retinopathy; the relation with incidence of nephropathy seems to be explained by an association with baseline albuminuria status concentrations, whereas the relation with incidence of proliferative retinopathy does not. PMID- 12774166 TI - Mortality from heart disease in a cohort of 23,000 patients with insulin-treated diabetes. AB - AIMS/HYPOTHESIS: Although ischaemic heart disease is the predominant cause of mortality in older people with diabetes, age-specific mortality rates have not been published for patients with Type 1 diabetes. The Diabetes UK cohort, essentially one of patients with Type 1 diabetes, now has sufficient follow-up to report all heart disease, and specifically ischaemic heart disease, mortality rates by age. METHODS: A cohort of 23,751 patients with insulin-treated diabetes, diagnosed under the age of 30 years and from throughout the United Kingdom, was identified during the period 1972 to 1993 and followed for mortality until December 2000. Age- and sex-specific heart disease mortality rates and standardised mortality ratios were calculated. RESULTS: There were 1437 deaths during the follow-up, 536 from cardiovascular disease, and of those, 369 from ischaemic heart disease. At all ages the ischaemic heart disease mortality rates in the cohort were higher than in the general population. Mortality rates within the cohort were similar for men and women under the age of 40. The standardised mortality ratios were higher in women than men at all ages, and in women were 44.8 (95%CI 20.5-85.0) at ages 20-29 and 41.6 (26.7-61.9) at ages 30-39. CONCLUSIONS/INTERPRETATION: The risk of mortality from ischaemic heart disease is exceptionally high in young adult women with Type 1 diabetes, with rates similar to those in men with Type 1 diabetes under the age of 40. These observations emphasise the need to identify and treat coronary risk factors in these young patients. PMID- 12774167 TI - Formation of insulin-positive cells in implants of human pancreatic duct cell preparations from young donors. AB - AIMS/HYPOTHESIS: Pancreatic ducts are considered as potential sites for neogenesis of beta cells. In vitro studies have reported formation of islets from postnatal human and rodent duct tissue. We examined whether postnatal human duct cell preparations can generate new beta cells after transplantation. METHODS: Pancreatic duct cells were prepared from the non-endocrine fraction of human donor pancreases that were processed for islet-cell isolation. Grafts containing 0.5 million duct cells with 1% contaminating insulin-positive cells were implanted under the kidney capsule of normoglycaemic nude mice. At 0.5 and 10 weeks post-transplantation, implants were examined for their cellular composition and for the volumes of their composing cell populations, i.e. cytokeratin 19 positive duct cells, synaptophysin-, insulin- and glucagon-positive endocrine cells. RESULTS: Between week 0.5 and 10, duct-cell volume decreased by at least 90% whereas the change in insulin-positive cell volume depended on donor age. Implants from donors over 10 years had a threefold decrease in their insulin positive cell volume, while those from donors under 10 years had a 2.5-fold increase. After 10 weeks, the implants from the younger donors consisted of 19% insulin-positive cells occurring as single units or small cell clusters. Three percent of these insulin-positive cells also expressed the ductal marker CK 19 and were consistently found in conjunction with ductal epithelia; up to 1% was positive for the proliferation marker BrdU and located in small endocrine cell clusters. CONCLUSIONS/INTERPRETATION: These data indicate that duct cell preparations from donors under 10 years can generate insulin-positive cells. This process might involve differentiation of CK 19-positive-insulin cells that are formed at the duct epithelia as well as proliferation of insulin-positive cells within endocrine cell aggregates. PMID- 12774165 TI - Diabetic dyslipidaemia: from basic research to clinical practice. AB - The recognition that the increase of plasma triglyceride rich lipoproteins (TRLs) is associated with multiple alterations of other lipoproteins species that are potentially atherogenic has expanded the picture of diabetic dyslipidaemia. The discovery of heterogeneity within major lipoprotein classes VLDL, LDL and HDL opened new avenues to reveal the specific pertubations of diabetic dyslipidaemia. The increase of large VLDL 1 particles in Type 2 diabetes initiates a sequence of events that generates atherogenic remnants, small dense LDL and small dense HDL particles. Together these components comprise the atherogenic lipid triad. Notably the malignant nature of diabetic dyslipidaemia is not completely shown by the lipid measures used in clinical practice. The key question is what are the mechanisms behind the increase of VLDL 1 particles in diabetic dyslipidaemia? Despite the advances of recent years, our understanding of VLDL assembly and secretion is still surprisingly incomplete. To date it is still unclear how the liver is able to regulate the amount of triglycerides incorporated into VLDL particles to produce either VLDL 1 or VLDL 2 particles. The current evidence suggests that the machinery driving VLDL assembly in the liver includes (i) low insulin signalling via PI-3 kinase pathway that enhances lipid accumulation into "nascent " VLDL particles (ii) up-regulation of SREBP-1C that stimulates de novo lipogenesis and (iii) excess availability of "polar molecules" in hepatocytes that stabilizes apo B 100. Recent data suggest that all these steps could be fundamentally altered in Type 2 diabetes explaining the overproduction of VLDL apo B as well as the ability of insulin to suppress VLDL 1 apo B production in Type 2 diabetes. Recent discoveries have established the transcription factors including PPARs, SREBP-1 and LXRs as the key regulators of lipid assembly in the liver. These observations suggest these factors as a new target to tailor more efficient drugs to treat diabetic dyslipidaemia. PMID- 12774168 TI - Fronto-orbital remodeling without orbito-naso-frontal bandeau. AB - INTRODUCTION: Fronto-orbital bilateral advance is the procedure of choice for the treatment of craniosynostosis affecting most of the anterior area of the skull and orbitomalar regions. The aim of the technique is to achieve a supra-orbital bilateral bar and a frontal bone. We have introduced a modification in order to simplify the technique. PATIENTS AND METHODS: From November 1998 to January 2002, 18 patients with craniosynostosis have been surgically treated using our technique. The mean age when the treatment was performed was 6.93 months (range 3 to 22 months). Brain computed tomography (CT) scans and three-dimensional (3-D) reconstruction of CT scans were performed before and after treatment. SURGICAL TECHNIQUE: A bifrontal craniotomy was performed taking the osteotomy up to the supraorbital rim. A new frontal bone was obtained from another region of the cranium creating new orbital edges. The osteosynthesis was conducted using absorbable materials. RESULTS: The follow-up of the patients ranged from 3 months to 3 years. All patients were studied using CT scans and 3-D reconstruction of CT after treatment, which demonstrated the persistence of the fronto-orbital advance. No secondary complications related to the new technique were found in any of the patients. CONCLUSIONS: The frontal-orbital advance obtained was stable. The technique was simplified by not creating a supraorbital bar and by reducing the bone fixation points. The manipulation of both frontal lobes and orbital globes was negligible. The aesthetic results were excellent. PMID- 12774169 TI - Short-term subarachnoid space drainage: a potential treatment for extraventricular hydrocephalus. AB - INTRODUCTION: Extraventricular hydrocephalus (EVOH), defined as the enlargement of all CSF compartments in the absence of an obstructing lesion, is usually associated with an increased head circumference and a relatively benign clinical course. Occasionally, because of concern about increased intracranial pressure (ICP), treatment with diuretics is initiated. In most cases, surgical drainage or diversion is not indicated. EVOH may follow a more alarming clinical course and be associated with developmental delay and/or other symptoms of increased ICP. CASE REPORT: We describe a 6-month-old girl with EVOH and developmental delay who was treated with temporary drainage of the subarachnoid space. Clinical response was immediate, with stabilization of the head circumference and improvement in motor performance. We propose that such a surgical procedure might be considered for more frequent use in selected cases. PMID- 12774170 TI - Endoscopic third ventriculostomy in tuberculous meningitis. PMID- 12774171 TI - Thermal physiology of the common eelpout (Zoarces viviparus). AB - We investigated the temperature dependence of some physiological parameters of common eelpout (Zoarces viviparus) from different locations (North Sea, Baltic Sea and Norwegian Sea) on acclimation temperature (3 degrees C and 12 degrees C) and acute temperature variation. The lethal limit of 12 degrees C-acclimated eelpout was determined as the critical thermal maximum [loss of equilibrium (LE) and onset of muscular spasms (OS)] and it was found to be 26.6 degrees C for LE and 28.8 degrees C for OS for all populations. However, these parameters do not have any relevant ecological interpretation. We therefore investigated the effect of gradually increased water temperature on standard metabolic rate (measured as resting oxygen consumption Mo2) and critical oxygen concentration ([O2]c) of eelpouts. Acclimation to low temperature (3 degrees C) resulted in partial compensation of Mo2, paralleled by a decrease of activation energy for Mo2 (from 82 kJ mol(-1) at 12 degrees C to about 50 kJ mol(-1) at 3 degrees C) in North Sea and Baltic Sea eelpouts. At the same time, Norwegian eelpout showed no acclimation of oxygen demand to warm temperature (12 degrees C) at all. The scope for eelpout aerobic metabolism shrank considerably with increased acclimation temperature, as [O2]c approached water oxygen concentrations. At 22.5+/-1 degrees C the [O2]c reached air saturation, which is equivalent to the upper critical temperature (TcII) and at this temperature the aerobic scope for the metabolism completely disappeared. In line with previous insight, the comparative analysis of the temperature dependence of Mo2 of Z. viviparus from different populations suggests that a pejus (sub-critical) temperature for this species is about 13-15 degrees C. In conclusion, the capacity to adjust aerobic metabolism relates to thermal tolerance and the bio-geographical distribution of the species. Global warming would thus be likely to cause a shift in the distribution of this species to the North. PMID- 12774172 TI - Experience-based plasticity of acoustically evoked aggression in a territorial frog. AB - Territorial male bullfrogs ( Rana catesbeiana) display lower levels of aggression toward familiar territorial neighbors compared to unfamiliar individuals based on the perception of individual differences in vocalizations. Stimulus-specific habituation elicited by repeated exposures to a neighbor's vocalizations is believed to play a role in mediating the low levels of aggression between neighbors. The present study describes habituation in multiple components of the bullfrog territorial aggressive response and examines whether these separate response components habituate at similar or different rates. In response to repeated broadcasts of synthetic bullfrog advertisement calls in a field playback experiment, the numbers of aggressive calls and movements, the latency to the first aggressive call, and the distance approached toward the playback speaker exhibited significant response decrements that developed at nearly identical rates. All four responses exhibited recovery upon subsequent broadcasts of a novel stimulus simulating a different individual. In contrast, the number of advertisement calls did not exhibit significant decrements with repeated playbacks. The patterns of stimulus-specific response decrement were inconsistent with explanations such as sensory adaptation, effector fatigue, non-specific changes in motivation, and a simple form of auditory habituation. Possible mechanisms for the observed plasticity in aggression are discussed in the context of anuran communication. PMID- 12774173 TI - Chronic prostatitis: what we know, what we do not know, and what we should do! AB - Chronic prostatitis is a major health problem in men. This review details the various types of prostatitis, discusses the possible aetiologies and potential pathogens. In addition, the symptoms and therapies are outlined. Those individuals affected by prostatitis suffer from a markedly reduced quality of life with, in many cases, only a limited benefit from treatment. Many questions remain unanswered, and more research is required in this significant disease. PMID- 12774175 TI - Mycobacterium marinum infections of the distal upper extremities: clinical course and imaging findings in two cases with delayed diagnosis. AB - Mycobacterium marinum infections cause tenosynovitis of the distal upper extremities and develop as a consequence of skin abrasions acquired in contaminated water. We report on two patients whose MR imaging studies showed tenosynovitis of the distal upper extremity secondary to M. marinum. In one patient sequential MR imaging showed development of bony erosions. Appropriate treatment was delayed in both patients because the diagnosis was not considered. We report on and discuss the clinical course and MR imaging findings in two patients with M. marinum infection. PMID- 12774174 TI - Etiology: where does prostatitis stop and interstitial cystitis begin? AB - Male chronic pelvic pain syndrome (MCPPS) and interstitial cystitis (IC) patients show striking similarities in clinical presentation, diagnostic evaluation, purported pathogenesis, and even response to therapy. This makes the distinction between them very difficult. This review aims to show just how easy it is for the clinician to miss the features of IC in the MCPPS patient (and conversely). Misdiagnosis can result in the rejection of various potentially helpful therapies while accurate early diagnosis will lead to appropriate therapies and a better quality of life for the patient. PMID- 12774176 TI - Achondroplasia and enchondromatosis in a female child. AB - We present a girl with achondroplasia and enchondromatosis. Coexistence of these two disorders has, to the best of our knowledge, not been reported previously. PMID- 12774177 TI - The involvement of ATP sulfurylase in Se(VI) and Cr(VI) reduction processes in the fission yeast Schizosaccharomyces pombe. AB - The response of Schizosaccharomyces pombe towards the oxyanions selenate [Se(VI)] and dichromate [Cr(VI)] was investigated in order to establish the involvement of the yeast ATP sulfurylase in their reduction. An ATP sulfurylase defective/selenate-resistant mutant of S. pombe (B-579 Se(R) -2) and an ATP sulfurylase-active/selenate-sensitive strain of S. pombe (B-579 Se(S)) were included in this study. The inhibitory effect of Se(VI) and Cr(VI) oxyanions on growth and bioaccumulation was measured. The sensitive strain showed natural sensitivity to selenate while the resistant mutant tolerated a 100-fold higher concentration of selenate. These results indicate that selenate toxicity to microorganisms is connected with the reduction of selenate to selenite. Both strains showed similar sensitivity to Cr(VI) and in this study there was no evidence that ATP sulfurylase participates in the reduction process of Cr(VI). PMID- 12774178 TI - Anaerobic oxidation of 2-chloroethanol under denitrifying conditions by Pseudomonas stutzeri strain JJ. AB - A bacterium that uses 2-chloroethanol as sole energy and carbon source coupled to denitrification was isolated from 1,2-dichloroethane-contaminated soil. Its 16 S rDNA sequence showed 98% similarity with the type strain of Pseudomonas stutzeri (DSM 5190) and the isolate was tentatively identified as Pseudomonas stutzeri strain JJ. Strain JJ oxidized 2-chloroethanol completely to CO(2) with NO(3)(- )or O(2) as electron acceptor, with a preference for O(2) if supplied in combination. Optimum growth on 2-chloroethanol with nitrate occurred at 30 degrees C with a mu(max) of 0.14 h(-1) and a yield of 4.4 g protein per mol 2 chloroethanol metabolized. Under aerobic conditions, the mu(max) was 0.31 h(-1). NO(2)(-) also served as electron acceptor, but reduction of Fe(OH)(3), MnO(2), SO(4)(2-), fumarate or ClO(3)(-) was not observed. Another chlorinated compound used as sole energy and carbon source under aerobic and denitrifying conditions was chloroacetate. Various different bacterial strains, including some closely related Pseudomonas stutzeri strains, were tested for their ability to grow on 2 chloroethanol as sole energy and carbon source under aerobic and denitrifying conditions, respectively. Only three strains, Pseudomonas stutzeri strain LMD 76.42, Pseudomonas putida US2 and Xanthobacter autotrophicus GJ10, grew aerobically on 2-chloroethanol. This is the first report of oxidation of 2 chloroethanol under denitrifying conditions by a pure bacterial culture. PMID- 12774179 TI - Molecular cloning and characterization of rainbow trout (Oncorhynchus mykiss) CCAAT/enhancer binding protein beta. AB - A full-length cDNA encoding CCAAT/enhancer-binding protein beta (C/EBPbeta) was cloned from rainbow trout by anchored PCR. The putative 291 amino acid protein has 53% and 32% identity to the zebrafish and Japanese flounder sequences, respectively, and 30-34% identity to tetrapod homologues. This clone contains most conserved C/EBPbeta domains except the second transactivation domain just like the zebrafish homologue. Also similar to zebrafish, rainbow trout produces only shorter C/EBPbeta isoforms (LAP and LIP) but not the longer isoform (LAP*). However, unlike the zebrafish and Japanese flounder homologues, trout C/EBPbeta has the short open reading frame (uORF) upstream of the start codon for LAP but in an alternate reading frame, a feature of tetrapod C/EBPbeta genes. In normal rainbow trout, C/EBPbeta mRNA was detected in peripheral blood leukocytes, head kidney, posterior kidney, liver, spleen, gills, intestine and muscle. RT-PCR revealed that transcript levels of trout C/EBPbeta are clearly higher in sodium alginate-induced peritoneal cells than in head kidney and peritoneal cells of saline-injected fish or head kidney cells of alginate-injected fish. Together with expression in immunologically important tissues, this indicates that C/EBPbeta is likely to be involved in the immune response just as in mammals. Southern hybridization suggested that C/EBPbeta is a single copy gene. There are no introns in this C/EBPbeta gene, just like the mammalian homologues. These data suggest that we have obtained the trout ortholog of C/EBPbeta. PMID- 12774180 TI - A refined method for creating saccular aneurysms in the rabbit. AB - We describe a refined animal model of human intracerebral aneurysms for testing endovascular devices for interventional neuroradiological procedures. Saccular aneurysms resulting from a stump of the right common carotid artery (CCA) were created in 15 New Zealand White rabbits by intraluminal incubation of elastase that was applied to the CCA after distal ligation of the CCA and proximal occlusion of the vessel using a pliable balloon. Subsequently a microcatheter was advanced to a position cranial to the balloon and the elastase was infused under fluoroscopic guidance to avoid retrograde flow to the trachea via aberrant vessels. Contrast-enhanced (CE) MRA at 1.5 T and conventional digital subtraction angiography was performed to test for aneurysm size, morphology and neck anatomy. In all 15 animals aneurysms resulted from the stump of the right CCA, ranging in size from 2.0 to 9.9 mm (mean 6.3 mm) in craniocaudal direction, 1.0 to 5.5 mm (mean 3.8 mm) in mediolateral direction and 1.0 to 3.8 mm (mean 2.4 mm) in neck diameter. Aneurysm morphology could be adequately demonstrated using CE MRA. On histological evaluation a loss of the internal elastic lamina was noted. The described method represents an easy, reliable, and reproducible method of aneurysm creation in the rabbit in an area of high shear stress. These aneurysms can be used for testing new endovascular devices for embolization of intracranial aneurysms. PMID- 12774181 TI - Endovascular treatment of basilar tip aneurysms associated with moyamoya disease. AB - We report the efficacy and safety of endovascular treatment of basilar tip aneurysms (BTA) in five patients with moyamoya disease. The patients underwent intra-aneurysmal embolisation with detachable platinum coils. Three BTA presented with subarachnoid haemorrhage (SAH); the other two were asymptomatic. In four cases, one embolisation procedure produced >95% angiographic obliteration of the aneurysm. In the other patient, 80-90% obliteration was achieved initially, but due to growth of the residual aneurysm, the procedure was repeated 7 months later. Two patients experienced transient oculomotor paresis as a procedure related complication. Mean follow-up was 43.6+/-34.0 months (range 8-92 months). One patient died of putaminal haemorrhage unrelated to the aneurysm 15 months after embolisation. The other four had no subsequent SAH and survived without sequelae. Endovascular embolisation using detachable platinum coils proved to be a safe and efficient treatment modality for BTA associated with moyamoya disease. PMID- 12774182 TI - Doppler sonograpy with dynamic testing in a case of aortic dissection extending to the innominate and right common carotid arteries. AB - A 50-year-old woman with Marfan's syndrome was admitted for an aortic dissection with an intimal flap extending from the sinus of Valsalva to the descending aorta and aortic valve incompetence. Ultrasonography revealed a double lumen in the innominate and right common carotid (RCCA) arteries. The false lumen extended from the aortic arch to the distal RCCA and compressed and nearly occluded the true lumen in the innominate artery. At the end of the RCCA was a large tear allowing communication between the false and true lumens. Colour-coded Doppler sonography showed blood passing from the false lumen into the true lumen and antegrade flow in the false lumen but reverse flow in the true channel. A dynamic test, as used in accessing for subclavian steal syndrome, producing reactive hyperaemia, showed the retrograde flow in the true channel to be markedly increased, supplying the subclavian artery. We emphasie the importance of functional description of an abnormal haemodynamic situation, which in this case helped to avoid unnecessary surgery to the supra-aortic arteries. PMID- 12774183 TI - Analysis of single mammalian cells with capillary electrophoresis. PMID- 12774184 TI - Nanoparticles in bioanalytics. PMID- 12774185 TI - Development, maintenance and temporal pattern of self-administration maintained by ecstasy (MDMA) in rats. AB - RATIONALE: +/-3,4-Methylenedioxymethamphetamine (MDMA; "ecstasy") use is increasing around the globe but there is a paucity of studies examining the abuse liability of this drug. OBJECTIVES: The ability of drugs to reinforce operant responding in laboratory animals is a valid and reliable predictor of abuse potential. MDMA is self-administered by humans, but there have been few reports of reliable self-administration by drug-naive laboratory animals. The present study sought to examine the acquisition and maintenance of MDMA self administration by laboratory rats. The influence of prior training with cocaine self-administration on the acquisition of MDMA self-administration was also examined. METHODS: MDMA self-administration (0.25-2.0 mg/kg per infusion) was examined in rats that were first trained to self-administer cocaine as well as by those that were drug-naive. The dose-dependency of MDMA self-administration and the temporal pattern of responding maintained by various doses of MDMA were examined. In some rats, self-administration of MDMA during a 24-h session was also examined. RESULTS: MDMA was self-administered by laboratory rats that were experienced with self-administration of cocaine as well as by rats that were initially drug naive. For drug naive rats, the acquisition of MDMA self administration (1.0 mg/kg per infusion) developed gradually during daily test sessions. The latency to acquisition of self-administration was shorter in cocaine-trained rats. Self-administration was dose-dependent, extinguished when saline was substituted for MDMA and, was reinstated when MDMA was reintroduced. During a 24-h self-administration session, a high rate of responding was produced during the first hour of the test session followed by a steady and lower rate of two to four responses per hour during subsequent hours of the test. CONCLUSIONS: These results suggest that prior experience with cocaine self-administration facilitates the acquisition of MDMA self-administration. The results also suggest that MDMA has abuse liability and that increased use of the drug should raise concern of a growing and widespread potential for chronic abuse. PMID- 12774186 TI - Operant responding for a visual reinforcer in rats is enhanced by noncontingent nicotine: implications for nicotine self-administration and reinforcement. AB - RATIONALE: Current conceptualizations of drug reinforcement assume that drug taking behavior is a consequence of the contingent, temporal relationship between the behavior and drug reward. However, stimulant drugs also potentiate the rewarding effects of other reinforcers when administered noncontingently. OBJECTIVES: These studies were designed to determine whether noncontingent nicotine enhances the reinforcing properties of a nonpharmacological reinforcer and whether this direct effect facilitates operant behavior within the context of a nicotine self-administration procedure. METHODS: Rats self-administered nicotine or food, or received noncontingent nicotine, saline, or food either with or without a response-contingent, unconditioned reinforcing visual stimulus (VS). RESULTS: Noncontingent nicotine, whether delivered as discrete injections based on a pattern of self-administered nicotine or as a continuous infusion, increased response rates maintained by the VS. There were no significant differences in responding by animals that received contingent compared with noncontingent nicotine when a VS was available. This increase was not observed in the absence of the VS or as a consequence of noncontingent food delivery. Operant behavior was equally attenuated and reinstated by the removal and subsequent replacement of contingent and noncontingent nicotine. Nicotine supported self-administration in the absence of response-contingent, nicotine-paired stimuli; however, response rates were drastically reduced compared with nicotine self-administration with the VS. CONCLUSIONS: Nicotine influences operant behavior in two ways: by acting as a primary reinforcer when it is contingent upon behavior, and by directly potentiating the reinforcing properties of other stimuli through a nonassociative mechanism. Nicotine self-administration and smoking may be largely dependent upon this later action. PMID- 12774187 TI - Ethical issues in the use of animals in biomedical and psychopharmocological research. AB - RATIONALE: The ethical debate concerning the use of animals in biomedical and pharmacological research continues to be replete with misunderstandings about whether animals have moral standing. OBJECTIVES: This article briefly reviews the central ethical positions and their relationship to the basic parameters of research regulation from an international perspective. The issues associated with the validation of animal models will then be discussed. Finally, suggestions for empirical ethics research will be presented. METHODS: Recent literature reviews were accessed and analyzed. RESULTS: This review summarizes the pertinent ethical and research literature. CONCLUSIONS: In summary, regardless of the ethical perspective one favors, there is strong agreement that animals matter morally and that at a minimum their welfare must be considered. This position is reflected in the structure of national regulatory schemes that emphasize the three Rs (replacement, reduction, refinement). Researchers should more actively participate in the discussion by becoming more knowledgeable about the details of the ethical issues. Research with animal models has been problematic in that it has often focused on attempting to produce global models of psychiatric disorders, which suffer from inherent validity problems. Researchers must also become more sophisticated about issues of model validation and the nature of the animals they use. PMID- 12774188 TI - Ethical considerations in psychopharmacological research involving decisionally impaired subjects. AB - RATIONALE: Research subjects who are unable to provide informed consent must be protected from exploitation. The federal regulations governing human subjects research mandate additional protections for "mentally disabled" subjects but include neither a definition of this "vulnerable" population nor any guidance on what safeguards should be employed or how they should be implemented. OBJECTIVES: This article begins with a definition of vulnerability due to a mental disorder in the context of the clinical research setting. It is organized along the following sequential phases of psychopharmacological research: study design and methodology; protocol review; subject recruitment and enrollment; conduct and monitoring of the study; and manuscript preparation and publication. Practical recommendations are then offered to clinical researchers and Institutional Review Boards (IRBs) for implementing additional protections for decisionally impaired subjects at each phase of the psychopharmacological research process. METHODS: A computer-assisted literature review was performed to identify descriptions of safeguards for decisionally impaired subjects. Recommendations for additional protections were also drawn from the authors' experiences with the IRB review process and the conduct and monitoring of clinical research with decisionally impaired subjects. RESULTS: The use of informed consent monitoring and the independent assessment of decision-making capacity are two feasible safeguards that IRBs can mandate for research with decisionally impaired subjects. There has been little systematic implementation of other safeguards such as research advance directives or prospective authorization for research participation. CONCLUSIONS: Clinical investigators and IRBs are under considerable scrutiny with respect to the protection of decisionally impaired research subjects. There is a pressing need for data-driven strategies for the optimal protection of decisionally impaired research subjects. PMID- 12774189 TI - Lorazepam should no longer be used as a prototypical benzodiazepine. PMID- 12774190 TI - Differences between d-methamphetamine and d-amphetamine in rats: working memory, tolerance, and extinction. AB - RATIONALE: Previously, we have shown that d-amphetamine (AMPH) was more potent than d-methamphetamine (METH) at increasing extracellular levels of dopamine (DA) in the prefrontal cortex (PFC) at doses that had similar effects in the nucleus accumbens. Since working memory depends on PFC DA, it was postulated that AMPH would also be more potent than METH at affecting working memory. OBJECTIVE: To determine if AMPH is more potent than METH at affecting working memory. METHODS: Working memory was measured in adult female Sprague-Dawley rats using a delayed alternation T-maze task with multiple delays (1, 10, 60 s) and food rewards. The percentage of food rewards consumed was also recorded. Animals were tested with METH and AMPH before and after a chronic protocol, with measurements of locomotor activity used to test for pharmacological tolerance or sensitization. The effects of METH and AMPH on extinction were also examined by omitting the food rewards from the T-maze. RESULTS. Both METH and AMPH produced dose-related bimodal effects on working memory at the intermediate delay (10 s); however, AMPH was more potent than METH. Both METH and AMPH initially also decreased the percentage of food rewards consumed in the T-maze. After chronic testing, animals displayed tolerance to both the working memory impairments and the reduction in food reward intake produced by AMPH. Animals did not display significant tolerance to the effects of METH on food reward consumption and performed worse in the T-maze after chronic testing. METH, but not AMPH, interfered with extinction. CONCLUSIONS: These results indicate that METH and AMPH differ in altering working memory and the expression of tolerance, perhaps due to differences in behavioral inhibition. PMID- 12774192 TI - Effects of Amanita phalloides toxins on insulin release: in vivo and in vitro studies. AB - The clinical picture of Amanita phalloides poisoning includes hypoglycaemia, usually related to hepatic damage. In fact, Amanita toxins induce hepatic glycogen depletion in humans and animals. However, in animals morphological changes of pancreatic beta cells are reported, suggesting that an alteration of insulin secretion might be involved in the pathogenesis of hypoglycaemia. Therefore, we determined fasting glucose, insulin and C-peptide levels in ten patients intoxicated by Amanita phalloides and in ten control subjects. Fasting blood samples were drawn on 3 consecutive days, beginning 48-72 h after mushroom ingestion, and glucose, insulin and C-peptide concentrations were determined by routine methods. Serum glucose concentrations did not differ between poisoned subjects and controls, whereas insulin and C-peptide concentrations were significantly higher in poisoned subjects ( P<0.01), with a significant positive correlation ( R=0.97, P<0.001). We also evaluated the effects of alpha-amanitin, the main amatoxin, on in vitro insulin release. Rat islets were incubated with 5 and 50 mg/l alpha-amanitin, in the presence or absence of 5.6 mM glucose. In another protocol, islets were preincubated for 2 h with 5 and 50 mg/l alpha amanitin in medium containing 5.6 mM glucose. After lavage, islets were incubated with increasing glucose (2.8-22.0 mM) to evaluate insulin release. In vitro, both concentrations of toxin induced insulin release (5 mg/l P<0.02, 50 mg/l P<0.01 vs controls), in the presence of 5.6 mM glucose. Islets preincubated with 5 mg/l alpha-amanitin showed a pattern of glucose-stimulated insulin release similar to controls, whereas islets preincubated with 50 mg/l alpha-amanitin showed an increased basal release with a reduced response to glucose stimulation. These observations show that Amanita toxins might play a role in the clinical context of Amanita poisoning. We demonstrate, for the first time, that alpha-amanitin induces insulin release and may exert a cytotoxic effect on beta cells. PMID- 12774191 TI - High levels of morphine-6-glucuronide in street heroin addicts. AB - RATIONALE: In the body, heroin is rapidly transformed to 6-acetylmorphine (6-AM) and then to morphine, that in turn is mainly metabolized to morphine-3 glucuronide (M3G) and, at lesser extent, to morphine-6-glucuronide (M6G). Unlike M3G, M6G is a potent opioid agonist. Intravenous heroin abusers (IHU) are exposed to a wide array of drugs and contaminants that might affect glucuronidation. OBJECTIVES: We assessed plasma and urine concentrations of M3G and M6G in four groups of subjects: the first two included long-term IHU either exposed to street heroin ( n=8) or receiving a single IV injection of morphine ( n=4), while the other two groups included non-IHU patients receiving acute IV ( n=8) or chronic oral ( n=6) administrations of morphine. METHODS: After solid phase extraction plasma and urine concentrations of morphine metabolites were determined by HPLC analyses. RESULTS: M3G accounted for the greater part of morphine glucuronides detected in body fluids of non-IHU patients treated with morphine. This pattern of metabolism remained stable across 15 days of oral administration of incremental doses of morphine. In contrast, the two groups of IHU (street heroin taking or morphine-treated subjects) showed a reduction of blood and urine M3G concentrations in favor of M6G. Consequently, M6G/M3G ratio was significantly higher in the two IHU groups in comparison with the non-IHU groups. CONCLUSIONS: Chronic exposure to street heroin causes a relative increase in concentrations of the active metabolite, M6G. Since the pattern of M6G action seems closer to heroin than to morphine, the increased synthesis of M6G observed in IHU may prolong the narrow window of heroin effects. PMID- 12774195 TI - Experimental analyses of body image in the chimpanzee. AB - It is difficult to study body image in animals. In this study, it is assumed that the perception of the body of others reflects body image. The perception of the human face was examined in a series of six experiments with a chimpanzee. Delayed matching-to-sample tasks were employed. Although the chimpanzee mastered the tasks and showed transfer of performance to new faces, subtle changes in the matching face resulted in the deterioration of performance. Responses of the chimpanzee were often controlled by factors other than the facial stimuli. Thus, although the chimpanzee has a body image as humans do, it may not be as clear and as segmented. PMID- 12774194 TI - Short-term calcitriol administration improves calcium homeostasis in adults with cystic fibrosis. AB - Osteoporosis is a well-defined health risk in cystic fibrosis (CF) patients due to many factors. Vitamin D insufficiency, despite routine cholecalciferol supplementation in CF patients, may contribute to a relative secondary hyperparathyroidism and possibly deficient bone mineralization. An alternate form of vitamin D, calcitriol, was studied to determine short-term effects on fractional calcium absorption and other calciotropic markers in 10 adult CF subjects and in 10 age-, sex- and body mass index (BMI)-matched controls. Serum fractional absorption of (45)Ca was determined after a calcium-containing meal prior to calcitriol intervention. Other measurements included serum parathyroid hormone (PTH), ionized calcium, 25-hydroxyvitamin D (25OHD), 1,25 dihydroxyvitamin D (1,25(OH)(2)D) and urinary calcium:creatinine and N telopeptide (NTx) concentrations. Both groups were then given calcitriol (0.5 micro g p.o. b.i.d. for 14 days) and restudied following the same protocol. Both groups increased their fractional absorption of (45)Ca after calcitriol ( p=0.015 CF subjects, p=0.001 controls), although calcitriol tended to be less effective in the CF group compared with the controls ( p=0.055). Post-prandial serum PTH concentrations were suppressed compared with baseline in both groups ( p=0.03 CF subjects, p=0.006 controls). Urinary NTx concentrations, a marker for bone resorption, decreased significantly in CF subjects after calcitriol (96.0+/-16.0 vs 63.9+/-12.7 nmol BCE/mmol Cr, p=0.01) and remained unchanged in the control group. The controls had an increase in serum 1,25(OH)(2)D concentrations (69.9+/ 4.2 vs 90.7+/-9.6 pmol/l, p=0.02) while there was no significant change in the CF group. Oral calcitriol administration appears to improve markers of calcium balance in adults with CF by increasing fractional absorption of (45)Ca and lowering PTH concentrations, similar to its known effects in healthy subjects, while also suppressing urinary NTx, a marker of bone turnover. PMID- 12774196 TI - Sequences affecting the regulation of solvent production in Clostridium acetobutylicum. AB - The high solvent phenotype of Clostridium acetobutylicum mutants B and H was complemented by the introduction of a plasmid that contains either an intact or partially-deleted copy of solR, restoring acetone and butanol production to wild type levels. This demonstrates that the solR open reading frame on pSOLThi is not required to restore solvent levels. The promoter region upstream of alcohol dehydrogense E (adhE) was examined in efforts to identify sites that play major roles in the control of expression. A series of adhE promoter fragments was constructed and the expression of each in acid- and solvent-phases of growth was analyzed using a chloramphenicol acetyl-transferase reporter system. Our results show that a region beyond the 0A box is needed for full induction of the promoter. Additionally, we show that the presence of sequences around a possible processing site designated S2 may have a negative role in the regulation of adhE expression. PMID- 12774197 TI - Fatal acute granulomatous pulmonary aspergillosis in a healthy subject after inhalation of vegetal dust. AB - A previously healthy 28-year-old man presented a few hours after inhalation of vegetal dust with acute bilateral community-acquired pneumonia, which caused death in 10 days despite treatment with broad-spectrum antibiotics, intravenous amphotericin B, inotropic support, and mechanical ventilation. A postmortem lung biopsy indicated miliary granulomatous pulmonary aspergillosis. Six similar previously published cases of acute granulomatous pulmonary aspergillosis are reviewed. This entity in adulthood may reveal a defect in neutrophil or macrophage function, such as late-onset chronic granulomatous disease. PMID- 12774199 TI - Stroke in young adults: epidemiology. AB - Stroke in the young is rather rare. The absolute proportion of juvenile stroke is strongly linked to the structure of the population. In western European countries, with a high prevalence of old and very old people, less than 5% of strokes occur in subject under 45 years. The incidence is about 10 in 100,000 in several Italian studies. In Italy, about 3,300 new cases of juvenile stroke occur each year. Higher incidence rates have been found in the United States and developing countries. Subarachnoid hemorrhage prevails in young cases, compared with the total population of stroke patients. In the subgroup of patients with cerebral infarction, inter-atrial septum defects and inherited thrombophilic disorders seem to play an important role. PMID- 12774200 TI - Cervical cerebral artery dissection: a multicenter prospective study (preliminary report). AB - Forty-nine cases of dissection of the internal carotid and vertebral arteries are reported in our prospective multicenter study of 35 men and 14 women, with a mean age of 46.77 years. (range 17-60 years). We evaluated etiology, clinical manifestations, investigative techniques, and treatment. Thirty-one patients had so-called spontaneous dissections, although in the remaining 18 minimal or obvious trauma was considered as the etiological factor. Headache and neck pain occurred in 32 patients (65.3%). Local neurological manifestations were present in 15 patients (30.6%) and ischemic cerebral symptoms were present in 41 patients (83.6%). The most-relevant of the diagnostic tools are duplex sonography, magnetic resonance angiography, and angiography. Anticoagulation with heparin followed by warfarin was the treatment of choice in most of our patients. Complete recovery is reported in 14 patients (28.5%); 41 patients showed cerebral ischemic symptoms, of which 13 (26.5%) had good functional recovery. In 28 (57.1%), the NIHSS score decreased from 6.68 to 3.31 during hospitalization. PMID- 12774198 TI - Prolonged paradoxical reaction of tuberculosis in an HIV-infected patient after initiation of highly active antiretroviral therapy. PMID- 12774201 TI - Central nervous system vasculitis. AB - Vasculitis is an inflammation of the vessel wall. It may be either primary or secondary. Primary vasculitis includes systemic vasculitis (large, medium, and small-vessel vasculitis) and localized vasculitis (isolated angiitis of the central nervous system and non-systemic vasculitic neuropathy). Secondary vasculitis may be present in connective tissue disorders or may be caused by infections, neoplasms, and substance abuse. Patients presenting with symptoms suggestive of vasculitis require brain neuroimaging, lumbar puncture, and angiography, but only biopsy allows a definite diagnosis. PMID- 12774202 TI - Emerging risk factors for ischemic stroke. AB - The strongest evidence of a relationship between homocysteine (Hcy) and risk of cerebrovascular disease has been provided by six prospective studies. The vascular risk was shown to be dose dependent for both fasting and postmethionine Hcy levels and statistically independent of traditional cardiovascular risk factors, although there was a multiple effect in the presence of smoking and hypertension. Recently, it was demonstrated that not only hyperHcy but also MTHFR polymorphism is an independent risk factor for dissection. Finally, preliminary data suggest that hyperHcy is a risk factor for the occurrence of cerebrovascular events (transient ischemic attack/stroke) in patients with atrial fibrillation. On the basis of these results, several intervention trials are ongoing to determine whether lowering Hcy levels with vitamin supplementation will reduce the recurrence of stroke. PMID- 12774203 TI - Embolism from the heart in the young patient: a short review. AB - Embolism from the heart causes about one-fifth of ischemic strokes and transient ischemic attacks. Although a potential embolic source may be found in up to one third of cases, which are actually caused by other mechanisms, in the young patient more than one source of embolism is less likely. The most-important embolic sources are non-rheumatic and rheumatic atrial fibrillation (AF), infective endocarditis, prosthetic heart valves, recent myocardial infarction, dilated cardiomyopathy, intracardiac tumors, and rheumatic mitral stenosis. Uncomplicated mitral valve prolapse should no longer be considered a cause of embolism from the heart to the brain; an additional disorder (i.e., gross mitral regurgitation, AF, infective endocarditis) must be present to suggest a cardioembolic origin of the stroke. PMID- 12774204 TI - Stroke in young people: imaging. AB - Stroke in young people is an unexpected event, whose causes are more heterogeneous than in the older population. Moreover, its diagnosis must exclude a large number of other diseases. Thereby, imaging plays an important role in the diagnostic approach of these patients. The aims of the neuroradiological work-up are to confirm that an acute ischemic lesion has occurred, to determine the location and the extent of the lesion, to verify the patency of major neck and intracranial arteries and to exclude eventual underling pathologies. In this sense, MRI offers the best sensitivity and spatial resolution. The more frequent causes of ischemic stroke in young people are considered and the correspondent characteristic imaging aspects are described. PMID- 12774205 TI - Percutaneous closure of patent foramen ovale: pathophysiology, indications, and technique. AB - The percutaneous closure of patent forame ovale (PFO) is currently performed in patients with cryptogenic stroke or paradoxical systemic embolism. The availability of new user friendly devices and the increase in knowledge of the pathophysiology, epidemiology, and follow-up of these patients has broadened the indications and drastically reduced the morbidity related to the intervention. This review considers the main pathophysiological and epidemiological features of PFO and discusses the indications and results of the intervention. PMID- 12774206 TI - Minimally invasive closure of patent foramen ovale through port-access surgery. AB - We present here our experience of patent foramen ovale closure through a mini invasive surgical technique. With the new surgical platform termed Heartport System, it is possible to install extra-corporeal circulation and to obtain cardioplegic arrest of the heart with endovascular techniques. This allows the operation of closure to be entirely performed through a micro-thoracotomy (Port access surgery) in the right 4th intercostal space. Sixty-one patients underwent this surgical correction and seventeen of them (26%) had a diagnosis of patent foramen ovale, 12 with an history of cerebrovascular accidents. They are all alive and well after a mean follow-up of 16 months, with no recurrence of the inter-atrial communication and free from new neurological events. The post operative course was uneventful with patients discharged on postoperative day 4. This technique shows a 100% efficacy, minimise surgical trauma, allows a quick recovery and offers excellent cosmetic results with no scarring. We believe that this procedure must be regarded as the "golden standard" for the closure of atrial septal defects PMID- 12774207 TI - Diagnostic criteria for Parkinson's disease. AB - The diagnosis of Parkinson's disease is not easy. Developments in basic research have indicated pathophysiological links among parkinsonian syndromes that are still classified as independent entities. On the other hand, genetic studies are dividing forms that fit into the clinical diagnosis of Parkinson's disease. The diagnostic criteria used in current practice are by no means satisfactory, but cannot yet be replaced by new comprehensive criteria based on laboratory evidence. PMID- 12774208 TI - Motor complications of Parkinson's disease. AB - Long-term treatment with levodopa in Parkinson's disease results in the development of motor complications, including drug failure, reduced duration of antiparkinsonian action (wearing off phenomenon), sudden shifts between under treated and over-treated states (on-off phenomenon), freezing and involuntary movements such as levodopainduced dyskinesia. These motor complications can sometimes be solved with changes in the drug regimen, particularly the addition of dopamine agonists and catechol-O-methyltransferase (COMT) inhibitors and/or changes in levodopa dose, formulation and number of doses. Amantadine and selegiline can also be helpful in reducing motor fluctuations. PMID- 12774209 TI - Cognitive and behavioral disturbances in Parkinson's disease. AB - The dysfunction of the striato frontal circuits that occurs in Parkinson's disease results in cognitive and behavioural problems as well as motor impairment. Depression is frequent and cognitive deficits also occur that progress with advancing illness, sometimes to subcortical dementia. Spread of Lewy bodies to the cortex, cholinergic and monoaminergic neurotransmission deficiency, or concurrent Alzheimer pathology may be the anatomical bases of dementia. PMID- 12774210 TI - Autonomic dysfunction in Parkinson's disease. AB - Autonomic dysfunction in patients with Parkinson's disease (PD) has been recognized since the original description by James Parkinson in 1817. Autonomic failure can be the clinical presentation of other diseases like pure autonomic failure (PAF) and multiple system atrophy (MSA). Both the central and peripheral autonomic nervous systems can be affected in PD. Rajput and Rozdilsky described cell loss and Lewy bodies within the sympathetic ganglia and antibodies to sympathetic neurons have been detected in PD patients. Lewy bodies can be seen in autonomic regulatory regions, including the hypothalamus, sympathetic (intermediolateral nucleus of the thoracic cord and sympathetic ganglia), and parasympathetic system (dorsal, vagal, and sacral parasympathetic nuclei). Lewy bodies were also found in the adrenal medulla and in the neural plexi innervating the gut, heart and pelvis. Symptoms of dysautonomia are variable, and include cardiovascular symptoms, gastrointestinal, urogenital, sudomotor and thermoregulatory dysfunction, pupillary abnormalities and sleep and respiratory disorders. They may represent a useful tool in the differential diagnosis of "atypical" or "complicated" parkinsonisms. PMID- 12774211 TI - Differential diagnosis of Parkinson's disease and atypical parkinsonian disorders by magnetic resonance imaging. AB - Magnetic resonance imaging (MRI) is not usually considered helpful for establishing the diagnosis of Parkinson's disease, but it is much more important to support the clinical diagnosis in atypical parkinsonian disorders. In multiple system atrophy with predominant parkinsonian features (MSA-P), MRI demonstrates putaminal abnormalities due to loss of neurons and gliosis and accumulation of iron in the posterior lateral part of the nucleus. When cerebellar features are present (MSA-C), pontine and cerebellar atrophy is seen with signal abnormalities that correspond to the distribution of the degenerative changes. In progressive supranuclear palsy, the main abnormality is atrophy of the midbrain. Mild-to moderate cerebral atrophy may be present, but more-marked asymmetrical atrophy in the posterior frontal and parietal regions contralateral to the side of the clinical manifestations is characteristic of corticobasal degeneration. PMID- 12774212 TI - Surgery of Parkinson's disease: inclusion criteria and follow-up. AB - Parkinson's disease (PD) is a progressive disturbances of movement that affects mainly the motor system. Prolonged pharmacological administration may result in insufficient control of symptoms and significant side effects. Deep brain stimulation (DBS), targeted at the STN, is a recent surgical procedure that, according to the symptoms response, allows modification of stimulation parameters; its effects are also reversible. In this paper management of surgical patients is reported. It includes patient selection, inclusion and exclusion criteria, postoperative clinical protocol. The evaluation rating scale such as UPDRS, Dyskinesias Rating Scale and Self-Reporting Questionnaire usually administrated on PD patients are analyzed. Surgical inclusion criteria are (1) idiopathic PD, (2) IV or V Hoehn-Yahr stage, (3) severe motor disability, and (4) no dementia or psychiatric abnormalities. Postoperative clinical protocol is analyzed and parameter of stimulation after surgery and at the follow up are reported. Generally DBS allows an improvement of rigidity and tremor; bradykinesia also improves with high frequency stimulation. Results obtained by continuous stimulation show a mean improvement of UPDRS of about 60% and a significant reduction in the drug intake. PMID- 12774213 TI - Deep brain stimulation for Parkinson's disease: the experience of the Policlinico San Paolo Group in Milan. AB - Thirty patients with idiopathic Parkinson's disease were treated with deep brain stimulation electrode in the subthalamic nucleus. After surgery, the patients' best mean Unified Parkinson's Disease Rating Scale (UPDRS III) scores (medictionOFF-stimulatorON versus preoperative medicationOFF) were 77+/-14% at 3 months ( n=20 patients) and 72+/-14% at 12 months follow-up ( n=16). The mean reduction in therapy (expressed in levodopa dose equivalents in mg) was 68+/-25% at 12 months. Postoperative complications were rare, mostly mild, and reversible. Therapeutic success depends on a multidisciplinary team approach, meticulous patient selection, including patients' cognitive, psychic, and behavioral status, and patient and family lifestyles. PMID- 12774214 TI - Deep brain stimulation for the treatment of Parkinson's disease: the experience of the Neurosurgical Department in Monza. AB - deep brain stimulation is a widely accepted surgical therapy for the symptomatic treatment of advanced parkinson's disease; high frequency chronic stimulation of the subthalamic nucleus proved its efficacy to control the major motor symptoms. In the neurosurgical department of Monza we treated 72 parkinsonian patients (November 1998-January 2003). One year follow-up results are: decrease of tremor 90%, hypertonous 56%, bradykinesia 70%, voice impairment amelioration 30%, mean total daily L-dopa intake reduced 58%. Freezing and balance did not ameliorate, some voice impairment and psychic derangement have been observed. Major surgical complications were: haemorrage (1 case - transient hemiparesis), infections (2 cases), pulmonary embolisation (1 case). To optimise the surgical results, careful clinical and instrumental selection of the patients are mandatory before surgery. PMID- 12774215 TI - The role of the IKAP gene polymorphisms in atopic diseases in the middle European population. AB - Over ten genome-wide screens and many candidate genes studies were performed worldwide to elucidate genetic factors involved in the pathogenesis of bronchial asthma and other atopic diseases. Results from these studies were often discordant, which might have reflected complexity and heterogeneity of these multifactorial diseases. Among a variety of other loci, specific variants of the gene for IKAP (IKK complex-associated protein) were shown to be associated with bronchial asthma in children in a Japanese study. To test the possible role of SNPs in the coding region of the IKAP gene in atopic asthma or other atopic phenotypes in a highly homogenous Czech population, a case-control study including 373 patients and 309 healthy control subjects was performed. There were no significant differences in the genotype and allele distributions for any of five SNPs in the IKAP gene (T819C, G2295A, A2490G, T3214A and C3473T) between patients with atopic asthma or other atopic diseases and healthy controls. These results suggest that the polymorphisms in the coding region of the IKAP gene are unlikely to contribute to atopic disease risk in the Czech population. PMID- 12774216 TI - Construction of a toroidal model for the magainin pore. AB - Magainins are natural peptides that selectively kill bacteria at concentrations that are harmless to animal cells. Due to a positive charge and distinct hydrophobic moment, magainins in the alpha-helical conformation interact favorably with bacterial membrane lipids. These interactions lead to the formation of large openings in the membrane and to the cell's death. The openings (toroidal pores) are supramolecular structures consisting of lipid and peptide molecules. A computer model of the pore in a bacterial membrane was constructed (see Figure) for the study of the molecular basis for magainin selectivity and specificity. Details of the construction and the preliminary equilibration of the pore model are given in this paper. PMID- 12774218 TI - Morphological types of arbuscular mycorrhizal fungi in roots of weeds on vacant land. AB - Morphological types of arbuscular mycorrhizal (AM) fungi in weeds of vacant land were examined in spring and autumn. In total, 33 plant species belonging to 28 genera in 13 families were examined. The number of plant species with Arum-type AM was higher than those with Paris- or intermediate types in both seasons. Thus, Arum-type colonization may be beneficial for fast-growing plant species on vacant land. There was a strong relationship between plant identity and AM morphological type, as the colonization types were mostly distinguished at the plant family level. PMID- 12774217 TI - Cyclometalated ruthenium(II) complexes as efficient redox mediators in peroxidase catalysis. AB - Cyclometalated ruthenium(II) complexes, [Ru(II)(C~N)(N~N)(2)]PF(6) [HC~N=2 phenylpyridine (Hphpy) or 2-(4'-tolyl)pyridine; N~N=2,2'-bipyridine, 1,10 phenanthroline, or 4,4'-dimethyl-2,2'-bipyridine], are rapidly oxidized by H(2)O(2) catalyzed by plant peroxidases to the corresponding Ru(III) species. The commercial isoenzyme C of horseradish peroxidase (HRP-C) and two recently purified peroxidases from sweet potato (SPP) and royal palm tree (RPTP) have been used. The most favorable conditions for the oxidation have been evaluated by varying the pH, buffer, and H(2)O(2) concentrations and the apparent second-order rate constants ( k(app)) have been measured. All the complexes studied are oxidized by HRP-C at similar rates and the rate constants k(app) are identical to those known for the best substrates of HRP-C (10(6)-10(7) M(-1) s(-1)). Both cationic (HRP-C) and anionic (SPP and RPTP) peroxidases show similar catalytic efficiency in the oxidation of the Ru(II) complexes. The mediating capacity of the complexes has been evaluated using the SPP-catalyzed co-oxidation of [Ru(II)(phpy)(bpy)(2)]PF(6) and catechol as a poor peroxidase substrate as an example. The rate of enzyme-catalyzed oxidation of catechol increases more than 10000-fold in the presence of the ruthenium complex. A simple routine for calculating the rate constant k(c) for the oxidation of catechol by the Ru(III) complex generated enzymatically from [Ru(II)(phpy)(bpy)(2)](+) is proposed. It is based on the accepted mechanism of peroxidase catalysis and involves spectrophotometric measurements of the limiting Ru(II) concentration at different concentrations of catechol. The calculated k(c) value of 0.75 M(-1) s(-1) shows that the cyclometalated Ru(II) complexes are efficient mediators in peroxidase catalysis. PMID- 12774219 TI - What are the problems in palliative care? Results from a representative survey. AB - The development of a standardised core documentation for palliative care was initiated in Germany in 1996. Results from previous evaluations have shown the wide variability of the documentation in participating units. A different documentation form was used in 2001 using free text entries to find out what problems palliative care specialists perceive in their patients. Fifty-five of the 83 palliative inpatient units in Germany (66% of the units) as well as one unit each from Austria and Switzerland, documented 1304 patients in the core documentation in 2001. Inpatient care was continued until death for 531 patients, 604 patients were discharged home and 169 patients were transferred to other places of care. Palliative care treatment effectively reduced mean physical symptom intensity. Mean intensities of psychological and social problems also were reduced although not as much as physical symptom load. Nursing problems were reduced for those patients discharged but not for those who died in the unit. Anxiety and depression were the most frequent psychological problems. Nursing problems were focussed on impairment of mobility and other activities of daily living such as washing, nutrition and drinking. Excessive distress on caregivers and the organisation of home care were the predominant social problems. In conclusion, this representative prospective survey with the majority of palliative care units in Germany showed the high effectiveness of symptom relief. Using the categories identified in this study, checklists were constructed and included in the documentation forms that are currently used for the core documentation project. PMID- 12774220 TI - Cardiovascular calcifications in pediatric patients receiving maintenance dialysis. AB - Cardiovascular disease is a major cause of morbidity and mortality in adult patients with end-stage renal disease receiving maintenance dialysis. Coronary artery calcifications (CAC) contribute to the high prevalence of cardiac disease and are associated with hyperphosphatemia, an elevated calcium-phosphorus product (CaxP), and prolonged time on dialysis. Chronic inflammation and malnutrition are also associated with an increased risk for development of cardiac calcifications. Young adults receiving maintenance dialysis develop cardiac calcifications at a degree out of proportion to healthy adults of the same age and gender. Many of these young adults initiated dialysis as children or teenagers. Risk factors associated with the development of CAC are also seen in the pediatric dialysis population. To date, reports of cardiac calcifications in pediatric patients receiving maintenance dialysis are limited to post-mortem studies. We present two pediatric patients with ANCA-positive vasculitis diagnosed with cardiac calcifications while receiving maintenance dialysis. Hyperphosphatemia and an elevated CaxP product were seen in both patients and probably contributed to the development of extraskeletal calcifications. In addition, both patients had an underlying systemic inflammatory disease and significant weight loss/malnutrition that may have contributed to the early and rapid onset of cardiac calcifications. PMID- 12774221 TI - Transient proximal tubular renal injury following Ecstasy ingestion. AB - Multiple renal adverse effects have been anecdotally reported with the ingestion of 3,4-methylenedioxymethamphetamine (Ecstasy), a widely used recreational drug. These side effects include acute renal failure, necrotizing vasculitis, and hyponatremia, the mechanisms for which are unknown. We report a case of transient acute proximal tubular injury and hyponatremia associated with Ecstasy use. An 18 year-old woman presented with new onset seizures and polydipsia. Her initial laboratory evaluation revealed hyponatremia (Na 117 mEq/L), polyuria (urine output >400 mL/h for several hours), renal glycosuria (blood glucose 120 mg/dL, urine glucose >1,000 mg/dL), and solute diuresis (urine osmolality 552 mOsm/kg H2O). Urine electrolyte values reflected a low tubular reabsorption of phosphorus (TRP) of 68.1% (expected TRP >85% at serum P 2.3 mg/dL) with an appropriate transtubular potassium gradient of 3.0 (serum K 3.7 mEq/L). Her hyponatremia was slowly corrected. A repeat TRP after 48 h had normalized to 86.5%, and her glycosuria resolved. An extensive toxin screen was later reported positive for Ecstasy. To our knowledge, this is the first example of an acute and transient proximal tubular injury with Ecstasy ingestion. This complication may become more apparent with increasing use of this drug. PMID- 12774222 TI - Renal papillary necrosis induced by naproxen. AB - A 17-year-old healthy girl was admitted to our hospital with diffuse abdominal pain and decreased oral intake of about 11 days duration. About a week prior to admission, she had taken naproxen, 250 mg four times a day for 4 days. Physical examination was normal except for diffuse abdominal tenderness on deep palpation. Investigations revealed high serum BUN (42 mg/dl) and creatinine (4.0 mg/dl). Serum electrolytes and complement (C3, C4) levels and urinalysis were normal. Antinuclear-antibody and anti-dsDNA were negative. Kidney biopsy revealed renal papillary necrosis, acute tubular necrosis, and focal interstitial nephritis. A diagnosis of nonoliguric acute renal failure due to naproxen nephrotoxicity was made. She received intravenous hydration, and oral steroids, which was gradually discontinued in 3 months. A follow-up at 4 months revealed normal renal function with a serum creatinine of 1.1 mg/dl, BUN 7 mg/dl, and normal urinalysis. The report highlights a need for caution while using naproxen or any other nonsteroidal anti-inflammatory drugs, even for a short duration. PMID- 12774223 TI - Use of mycophenolate mofetil in steroid-dependent and -resistant nephrotic syndrome. AB - Cyclosporin (Cs-A) is an effective treatment for difficult cases of nephrotic syndrome (NS), but its use can be complicated by renal toxicity and a high incidence of relapses after withdrawal. We reviewed the charts of 10 Cs-A dependent patients and 4 patients with steroid-dependent nephrotic syndrome (SDNS) not previously treated with Cs-A therapy. All patients had persistent NS, even after prior treatment with oral cyclophosphamide. Of 10 patients treated with Cs-A, 4 had surveillance renal biopsies consistent with Cs-A toxicity, and 8 of 10 had interstitial fibrosis prior to mycophenolate mofetil (MMF). Patients were treated with MMF, at 1,200 mg/m(2) per day, in an attempt to allow weaning of Cs-A and/or steroid therapy, and reduce the frequency of relapses. Overall, a significant decrease in frequency of relapses was noted after initiation of MMF therapy. In addition, 5 patients were weaned off Cs-A by 1-2 years of follow-up. One patient was weaned off Cs-A and MMF, and remained in complete remission. However, the subgroup of patients with frequently relapsing SDNS not treated with Cs-A appeared to have a reduction in the number of relapses while on MMF that did not reach statistical significance. Two patients with intractable steroid resistant NS continued to relapse repeatedly on MMF and Cs-A therapy. We conclude that in this small, single-center, uncontrolled experience, MMF therapy in patients with Cs-A-dependent NS appears to be effective in reducing Cs-A exposure. In addition, MMF appears to significantly decrease the frequency of relapses in this patient population. Further controlled studies are warranted to better define the potential efficacy and side effects of long-term MMF therapy in this setting. PMID- 12774224 TI - Are impacts of an exotic predator on a stream food web influenced by disturbance history? AB - Predatory species have been introduced to habitats spanning a wide range of environmental conditions. To better understand the consequences of predation in natural communities we need to examine how variations in abiotic factors modify the influence of predation. The effects of introduced predators may vary amongst habitats if natural disturbance affects the abundance and taxonomic composition of consumers and their resources, or the predator alters recolonisation after disturbance. We tested whether a bed-moving disturbance altered subsequent interactions involving native and introduced predatory fish, invertebrate grazers and algae in experimental channels within a New Zealand stream. Disturbance reduced the abundance of invertebrates by 84%, and induced mortality of Conoesucidae caddisflies. However, the relative abundance of taxa changed little immediately following the disturbance. Invertebrate communities recovered following disturbance in fishless channels and those with native galaxiids (Galaxias vulgaris), and were almost indistinguishable from undisturbed fishless controls after 2 weeks. Invertebrate abundance declined and algal abundance increased in channels with exotic brown trout (Salmo trutta) and their effect was strongest in previously disturbed channels. However, predators and disturbance only had interactive effects on grazer emigration rates. Trout affected grazers through direct consumption (e.g. Conoesucidae caddisflies), and induced higher emigration rates of grazers from channels via drift (e.g. the mayfly Deleatidium). The effects of predatory trout and galaxiids combined differed in disturbed and stable channels. The observed combined effects of predatory trout and galaxiids on invertebrate grazers were lower than expected in stable channels partly due to low emigration rates of Conoesucidae, whereas emigration of grazers was higher than expected in the disturbed channels. The biomass of algae was higher than expected in disturbed channels with both predators. Collectively, our results indicate that predator substitutability and the non-lethal effects of introduced predators varied depending on disturbance history, but their effects on the biomass of grazers and algae did not. PMID- 12774225 TI - Energetics of bluetongue lizards (Tiliqua scincoides) in a seasonal tropical environment. AB - We studied the physiological ecology of bluetongue lizards (Tiliqua scincoides) on the Adelaide River floodplain in tropical Australia to determine the seasonal patterns of energy expenditure and to determine the mechanisms by which seasonal differences were achieved. Field metabolic rates (FMR) were significantly lower in the dry season (37.6 kJ kg(-1) day(-1); n=9) than in the wet (127.3 kJ kg(-1) day(-1); n=7). Water flux was also lower in the dry season (6.8 ml kg(-1) day( 1); n=9) than in the wet (39.4 ml kg(-1) day(-1); n=7). Measurements of body temperatures (T(b)) and movements of free-ranging animals, and standard metabolic rate (SMR) of recently caught animals, allowed a detailed analysis of energy budgets for wet and dry seasons. In the dry, bluetongue lizards expended 90 kJ kg(-1) day(-1) less energy than in the wet season. Unlike some other lizards of the wet-dry tropics, SMR did not differ between seasons. About 5% of the seasonal difference in FMR was due to lower night time T(b) during the dry season, and about 7% was due to lower diurnal T(b). The remaining 88% of the decrease in energy expended in the dry season was due to a substantial decrease in other costs that may include reproduction, growth, digestion and activity. If we assume the animals fed daily and the costs of digestion are taken into account, the estimates are: 14% of the savings result from lower T(b) at night, 20% from lower T(b) in the day, and 66% result from decreased activity. It is therefore apparent that, unlike some agamid and varanid lizards that use a combination of behavioural and physiological mechanisms to conserve energy when food and water are limited, bluetongue lizards primarily use behavioural mechanisms to achieve a dramatic reduction in energy expenditure in the dry season. PMID- 12774226 TI - Spatial analysis of stable isotope data to determine primary sources of nutrition for fish. AB - Carbon and nitrogen stable isotopes were used to determine the ultimate autotrophic sources supporting production of three commercially important fish species over unvegetated mudflats in a subtropical estuary. Mean isotope values over the whole estuary for fish and autotroph sources were modeled to indicate feasible combinations of sources. Variability in isotope values among nine locations (separated by 3-10 km) was then used as a further test of the likelihood that sources were involved in fish nutrition. A positive spatial correlation between isotope values of a fish species and an autotroph indicates a substantial contribution from the autotroph. Spatial correlations were tested with a newly developed randomization procedure using differences between fish and autotroph values at each location, based on carbon and nitrogen isotopes combined in two-dimensional space. Both whole estuary modeling and spatial analysis showed that seagrass, epiphytic algae and particulate organic matter in the water column, including phytoplankton, are likely contributors to bream (Acanthopagrus australis) nutrition. However, spatial analysis also showed that mangroves were involved (up to 33% contribution), despite a very low contribution from whole estuary modeling. Spatial analysis on sand whiting (Sillago ciliata) demonstrated the importance of two sources, mangroves (up to 25%) and microalgae on the mudflats, considered unimportant based on whole estuary modeling. No spatial correlations were found between winter whiting (Sillago maculata) and autotrophs, either because fish moved among locations or relied on different autotrophs at different locations. Spatial correlations between consumer and source isotope values provide a useful analytical tool for identifying the role of autotrophs in foodwebs, and demonstrated here that both in situ production of microalgae and organic matter from adjacent habitats were important to fish over mudflats. PMID- 12774227 TI - The putative sensor histidine kinase CKI1 is involved in female gametophyte development in Arabidopsis. AB - Embryo sac formation is a fundamental step in sexual reproduction in plants. However, the key players involved in the development of the female gametophyte remain elusive. We present data indicating that a two-component sensor histidine kinase, CKI1, originally implicated in cytokinin perception, is required for completion of megagametogenesis in Arabidopsis. We isolated a loss-of-function mutation in CKI1 resulting from an insertion of the En-1 transposon into the CKI1 coding sequence. Genetic analysis revealed that the mutant allele, cki1-i, could not be transmitted through the female germ line. Confocal laser scanning microscopy identified a block in megagametogenesis, characterized by the abortion of the central vacuole in mutant embryo sacs, and degradation of the developing female gametophyte after completion of all mitotic divisions. The recovery of two independent stable alleles and one revertant wild-type allele resulting from En-1 excision confirmed unambiguously the causal link between the cki1-i mutation and the abnormal phenotype. In situ localization of CKI1 mRNA and histochemical analysis of stable transformants harboring the uidA gene under the control of CKI1 promoter revealed that expression of CKI1 starts at the very beginning of female gametophyte development, and continues until fertilization. This suggests that the developing embryo sac may remain sensitive to signals recognized by CKI1 throughout megagametogenesis. Furthermore, expression of the paternally transmitted CKI1 was detected early after fertilization. The results indicate a role for a two-component signaling system during female gametophyte development, and provide the first evidence that gametophytic expression of a sensor-like molecule is essential for specific processes during megagametogenesis. PMID- 12774228 TI - Cystic echinococcosis in Jordan: socioeconomic evaluation and risk factors. AB - The costs of illness and surgical intervention for human cystic echinococcosis (CE) cases in Jordan was economically evaluated by 77 surgeons and 77 CE patients. The cost of diagnosis for each CE case was 111.30 US Dollars and 146.20 US Dollars as estimated by surgeons and patients, respectively. The cost of surgical extraction of hydatid cysts for each case was 590.20 US Dollars and 638.50 US Dollars as estimated by both groups, respectively. Knowledge, attitudes and practices (KAP) of 77 CE patients as well as several Jordanian groups with different occupations including 144 shepherds, 119 settled livestock owners, 25 slaughter house workers, 400 university students and 80 inhabitants of a CE focus in southern Jordan were analyzed through a set of questionnaires. All of these groups had poor knowledge of CE, especially the source and causes of infection. All practices and attitudes of each group favored continuous transmission of the parasite and indicate the need for the implementation of a proper control program in the country. PMID- 12774229 TI - Echocardiographic findings in children ill with acute postinfectious glomerulonephritis. AB - There are many studies about heart dysfunction and its significance in chronic renal diseases, but only few data regards cardiac involvement in acute glomerular diseases. The aim of this study was to assess echocardiographic changes in children with acute postinfectious glomerulonephritis (APGN), and the relationship of those changes to blood pressure (BP), edema and glomerular filtration rate (GFR). Echocardiography was performed on 127 patients on admission to the hospital, on a random sample consisting of 51 patients after 6-8 weeks, and 124 controls. In the acute phase, APGN patients had a greater left ventricular (LV) internal end-diastolic diameter (LVIDd) (P=0.022), interventricular septum thickness (IVSd) (P=0.038), LV mass (LVM) (P=0.0001), longer early diastolic flow deceleration time (DT) (P=0.0001), and higher numbers of cases with mitral regurgitation (MR) (P=0.0001) and pericardial effusion (P=0.0001) in comparison with the controls. Changes were more evident in the youngest patients. At follow-up after 6-8 weeks, echocardiographic parameters significantly improved, but LV posterior wall thickness, IVSd, LV end-diastolic volume, LVM and DT remained greater than in the controls. GFR and edema influenced echocardiographic parameters. In conclusion, in a group of children ill with APGN, increases in left ventricular dimensions, wall thickness, LVM and changes in the left ventricular diastolic function related to edema and GFR were found. Whether different treatment regimens effect these changes requires further study. PMID- 12774230 TI - Expression pattern of a basic helix-loop-helix transcription factor Xhairy2b during Xenopus laevis development. AB - We report on the temporal and spatial expression pattern of the Xenopus laevis hairy2b ( Xhairy2b) transcription factor. Xhairy2b transcripts are present maternally, and expressed throughout the prospective ectoderm prior to the gastrula stage. During gastrulation, Xhairy2b expression is restricted to the deep layer of the Spemann organizer and three distinct regions in the prospective neuroectoderm, neural plate border, notoplate and anterior neural plate. At later stages, Xhairy2b expression is localized to prechordal plate, presomitic mesoderm, neural tube, neural crest derivatives and several tissue territories of the central nervous system. The analyses of Xhairy2b and several other hairy related genes suggest potential roles for Xhairy2b in the formation of boundaries in neural tissue territories. PMID- 12774231 TI - Assembling a task space: global determination of local shift costs. AB - Most theorizing on the origin of the costs that are associated with task shifts focuses on local transitions between individual trials. In the present article we argue that this emphasis has resulted in a neglect of more global representational structures, which also determinate shift costs. To substantiate this claim, we employed a set of four tasks that results from a factorial combination of two types of judgment and two judgment-to-response mappings. From previous work it is known that this kind of task combination is associated with a characteristic profile of shift costs as a function of the relation between successive tasks. Previously we have interpreted this profile as an indication of a hierarchically ordered dimensional representation of the two types of judgment and the two judgment-to-response mappings. Such a representation can only be expected to emerge when the two task features, judgment and mapping, vary independently of each other within the same situation. It is shown that the characteristic shift cost profile can indeed only be observed when the performance of any of the four tasks is required in a block of trials. In contrast, with only two tasks occurring in each block of trials, shift costs do not reflect the relation between successive tasks. This finding confirms the importance of global representational structures as a determinant of shift costs beyond local transitions between individual trials. PMID- 12774232 TI - A novel approach to in situ characterization of pancreatic beta-cells. AB - The tissue-slice technique has enabled major insights into neural and neuroendocrine physiology. Our aim was to adapt this technique to study the function of the endocrine pancreas. The preparation combines an in situ approach, as in gland perfusion, with a resolution characteristic of electrophysiological studies on single cells. The membrane potential in beta-cells in the slices recorded using the whole-cell patch-clamp was close to the calculated reversal potential for K+. With sufficient ATP in the recording pipette the beta-cells depolarized rapidly on exposure to an increased glucose concentration or stimulation with tolbutamide. The cells preserved bursting and spiking capacity for tens of minutes despite the whole-cell dialysis. In addition, the voltage clamp was used to monitor the changes in the membrane capacitance and to allow correlation of the electrical activity and the cytosolic calcium changes. The pancreatic tissue slice preparation is a novel method for studying the function of the beta- and other pancreatic endocrine and exocrine cells under near physiological conditions. PMID- 12774233 TI - The effect of concomitant vascular disruption in patients with iatrogenic biliary injuries. AB - BACKGROUND AND AIMS: To evaluate treatment results in iatrogenic biliary injuries with concomitant vascular injuries. PATIENTS/METHODS: Between January 1998 and May 2002 (inclusive), angiography was performed in 45 of the 105 patients treated for iatrogenic biliary tract injury. The charts of these 45 patients and 5 other patients in whom vascular injury was diagnosed at operation were evaluated retrospectively. Twenty-nine patients had concomitant vascular injury, the biliovascular injury group (BVI), and the remaining 21 patients had isolated biliary tract injury (IBTI). RESULTS: The most frequent initial operation was a cholecystectomy. The frequency of high-level (Bismuth III or IV) strictures was 90% in the BVI group and 62% in the IBTI group ( P<0.05). Perioperative mortality was 7% in the BVI group and 5% in the IBTI group ( P>0.05). The morbidity in the BVI group was significantly higher ( P<0.05). Two patients in each group were lost to follow up. During a median (range) follow up of 31 months (5-51 months), a successful functional outcome was achieved in 96% of the BVI group and 100% of the IBTI group with a multimodal approach ( P>0.05). CONCLUSIONS: The frequency of high-level biliary injury and morbidity were significantly higher in the BVI group. However, concomitant vascular injury had no significant effect on mortality and medium-term outcome of biliary reconstruction. Thus, routine preoperative angiography is not recommended. PMID- 12774234 TI - Deep neck infection: a present-day complication. A retrospective review of 83 cases (1998-2001). AB - Deep neck infections are less and less frequent today than in the past. Nevertheless, their complications are often life-threatening. The present study reviews the experience of the Department of Otolaryngology and Head and Neck Surgery of Padua with deep neck infections during the period from 1998 to 2001. Eighty-three patients (55 males and 28 females) were retrospectively considered. The site of origin of deep neck infection was identified in 76 patients (91%). The most common cause was dental infection, occurring in 35 cases (42%). In 12 cases (14%) deep neck infection was a complication of oropharyngeal infection. The relatively high incidence of Peptostreptococcus sp, Streptococcus viridans, Streptococcus intermedius and constellatus isolation was consistent with the high rate of odontogenic cases. Surgery was advocated as the treatment for any infection of the deep neck spaces. The recent series has demonstrated that medical treatment did not seem to increase complication rates or mortality. Our tailored approach (medical or medical and surgical) based on clinical and radiological evidence was successful in 97% of the patients. PMID- 12774235 TI - Autofluorescence videoendoscopy for photodiagnosis of head and neck squamous cell carcinoma. AB - The diagnosis of head and neck squamous cell carcinoma is usually made by visual identification. Searching for a non-invasive optical diagnostic method with the ability to detect the precancerous lesions or second primary tumors earlier in high-risk populations led to the development of photodiagnosis by autofluorescence (AF) endoscopy. The aim of the present study was to evaluate and discuss the diagnostic potential of autofluorescence videoendoscopy as a complementary visual aid in the routine endoscopic diagnosis of head and neck cancers. In a prospective study, 48 patients underwent white light (WL) videoendoscopy followed by AF technique at the Institute of Gustave-Roussy from November 2001 to August 2002. Of 48 patients, 30 had suspected precancerous or cancerous laryngeal lesions, 7 presented benign laryngeal lesions, while 8 showed pharyngeal and 3 oral tumors. All detected lesions were evaluated by histological examination. AF was induced by filtered blue light of a xenon short arc lamp and processed by a CCD camera system (D-Light AF System; Storz, Tuttlingen, Germany). Normal laryngeal mucosa displayed a typical green fluorescence signal. Moderate and severe epithelial dysplasia, carcinoma in situ and invasive carcinoma showed a diminished green fluorescence and presented a marked reddish-blue color. In case of hyperkeratosis a bright white color was detected. Some benign lesions, such as granulomas, polyps and papillomas also displayed altered green fluorescence. Autofluorescence videoendoscopy for photodiagnosis of head and neck squamous cell carcinomas has proved to be a method of high specificity and good sensitivity. Two additional precancerous lesions that were invisible at the WL examination but detected with the AF technique show its potential role in the regular screening procedure or follow-up examination in a high-risk population. It was a very helpful complementary visual aid for the intraoperative control of the surgical margins after per oral endoscopic resection. PMID- 12774236 TI - Three-dimensional versus two-dimensional ultrasound measurement of follicular volume: are they comparable? AB - To compare the accuracy of three-dimensional (3-D) versus two-dimensional (2-D) ultrasonography (U/S) in the measurement of follicular volume and identification of cumulus oophorus complex, thirty women were selected. Women who undergoing ovarian stimulation for in-vitro fertilization and embryo transfer (IVF-ET) using the long protocol of GnRH agonist were scanned by 2-D and 3-D ultrasonography before follicular aspiration. The volume of aspirated follicular fluid was compared with corresponding volume measured by 3-D and 2-D ultrasonography. It was found that, the high accuracy of 3-D U/S measurement of follicular volume is demonstrated clearly by limits of agreement which were within 1 ml of the true volume. While, the 2-D U/S produced limits of agreement those were up to 3.5 ml above or 2.5 ml below the true volume. Also, 3-D U/S could identify cumulus oophorus complex in 86% of cases. From this study, it was concluded that 3-D U/S system is more accurate than 2-D U/S in the measurement of the true follicular volume. Also, 3-D U/S is superior to 2-D U/S for identification of cumulus oophorus complex. PMID- 12774237 TI - Primary ovarian trabecular carcinoid tumour: a case report with an immunohistochemical study and a review of the literature. AB - INTRODUCTION: Primary ovarian carcinoid tumours are uncommon neoplasias. There are distinct histological types with different behaviours: insular, trabecular, mucinous and mixed. The trabecular subtype is very rare and unlike other carcinoid subtypes, it is characterised by the absence of a clinical carcinoid syndrome and has been related with a better prognosis than the others. No distant metastases have yet been reported. CASE REPORT: We present a case of a 76-year old woman diagnosed with a left ovarian tumour. She underwent a radical hysterectomy with bilateral salpingo-oophorectomy and regional lymph node clearance. Histology revealed an ovarian trabecular carcinoid tumour and no adjuvant treatments were performed. The patient is alive and free of disease 70 months following diagnosis. DISCUSSION: Immunohistochemical staining for p53 protein, and a comparative study with other subtypes of ovarian carcinoid tumours (insular and mucinous) is discussed in this report. PMID- 12774238 TI - Monteggia type I equivalent lesion: diaphyseal ulna and radius fractures with a posterior elbow dislocation in an adult. AB - BACKGROUND: A rare type I Monteggia equivalent lesion with a posterior dislocation associated with a diaphyseal radius and ulna fracture in an adult is described. The probable mechanisms of injury are speculated to include flexion of the elbow and pronation of the forearm. METHODS AND RESULTS: Early reduction of the dislocation and rigid fixation of the fractures helped to achieve excellent results. PMID- 12774239 TI - Increased expression of neurotrophins and their receptors in the mechanically compressed spinal cord of the spinal hyperostotic mouse (twy/twy). AB - The purpose of the present study was to identify any compensatory changes at the site of chronic compression of the spinal cord and neighboring segments. For this purpose, serial immunohistochemical and immunoblot analyses were performed for the expression levels of endogenous brain-derived neurotrophic factor (BDNF), neurotrophin (NT)-3, and their receptors, trkB and trkC in 24 tip-toe walking Yoshimura mice (twy/twy) aged 12-24 weeks. The twy mouse exhibits spontaneous calcified deposits posteriorly at the C1-C2 level, compressing the spinal cord. Immunoreactivities for BDNF, NT-3, trkB and trkC were preferentially localized in the gray matter, particularly in the anterior horn cells. In 24-week-old twy mice with severe compression, expression levels of these neurotrophins at the site of maximal compression were significantly lower than at the less- or non-compressed sites. In contrast, the expression levels of BDNF, NT-3, trkB and trkC were significantly higher at the rostral and caudal sites immediately adjacent to the maximal compression site. No such changes were noted in 12-week-old twy mice or in control Institute of Cancer Research mice. Our results suggest that overexpression of BDNF, NT-3, trkB and trkC in motoneuron areas neighboring the site of mechanical compression may represent compensatory changes in response to the compromised neuronal function at the level of compression, and that these proteins possibly contribute to neuronal survival and plasticity. PMID- 12774242 TI - Cyclo-oxygenase 2 inhibitors: emerging roles in the gut. AB - BACKGROUND AND AIMS: Discovery of an isoform of Cyclo-oxygenase (COX) 1, the inducible COX-2, has made it possible to avoid some side effects of non-specific COX inhibitors. The COX-2 gene is over-expressed in reflux oesophagitis, Barrett's oesophagus, gastric and colon cancer, familial adenomatous polyposis, pancreatic cancer, hepatocellular carcinoma, hepatotoxicity, cirrhosis, and inflammatory bowel disease, and specific COX-2 inhibitors have been tried experimentally and clinically and found effective. METHODS: A Medline search was performed of English-language experimental studies and controlled clinical trials from January 1980 to January 2002, and relevant citations were noted. RESULTS: Review of available literature shows that sulindac and COX-2 inhibitors are effective in preventing as well as regressing familial adenomatous polyposis. However, they have not been shown to prevent cancer in these patients. Studies evaluating NSAIDs and COX-2 inhibitors in carcinogen-induced and genetically manipulated animal models of various cancers have been promising especially in conditions such as Barrett's oesophagus, oesophageal and hepatocellular carcinoma and pancreatic cancer. COX-2 inhibitors may be of value in the treatment of reflux oesophagitis, pancreatitis and hepatitis, although carefully planned randomized controlled clinical trials demonstrating their efficacy need to be conducted. At present NSAIDs and COX-2 inhibitors cannot be recommended for average-risk individuals or for those with sporadic colorectal neoplasia (or other forms of cancers) as chemo-preventive agents. CONCLUSION: COX-2 inhibitors may open up a new therapeutic era in which these drugs can be used for chemo prophylaxis. However, COX-2 selective inhibitors retain renal adverse effects of the non-selective inhibitors and the concern regarding the pro-thrombotic potential of COX-2 inhibitors will limit their value as chemo-preventive agents. PMID- 12774240 TI - Resistance mechanisms of gastrointestinal cancers: why does conventional chemotherapy fail? AB - BACKGROUND: Gastrointestinal cancers belong to the most important causes of cancer death in the Western world. Because cure can be achieved only by complete surgical removal of the tumor, and most patients have metastasis at the time point of diagnosis, the majority of patients receive chemotherapy. DISCUSSION: Indications for chemotherapy are either the prevention of recurrence after tumor resection (neoadjuvant or adjuvant) or palliative treatment if the tumor is already widespread at diagnosis. Although gastrointestinal cancers often respond to primary treatment, the long-term results are disappointing. This is attributable to a variety of cellular resistance mechanisms, namely: (a) kinetic resistance due to slow growth rates that preclude the use of topoisomerase IIalpha inhibitors and related drugs; (b) genetic resistance due to mutations, for example, in the p53 gene, which impede the sensing of DNA damage and obstruct apoptotic pathways; (d) pharmacokinetic resistance, due to an excess of target proteins, inadequate drug metabolism, administration period, time or drug interactions; and (d) biological resistance due to tumor-induced environmental changes. These factors interfere specifically with the molecular mode of action of standard drugs used in the therapy of gastrointestinal cancers. CONCLUSION: Awareness of the various causes of drug resistance may help to devise individual tumor-adapted treatment designs. Notably, nonsteroidal antiphogistics may delay carcinogenesis, anticoagulants may increase the vulnerability of circulating tumor cells and reduce the nesting abilities of single tumor cells, inhibitors of angiogenesis may quell the growth of micrometastases, and kinase inhibitors may be administered as sensitizers to cytotoxic treatment. PMID- 12774243 TI - Binding of gastrointestinal tumor cells to endothelial E- and P-selectin adhesion receptors leads to transient down-regulation of sLeX ligands in vitro. AB - BACKGROUND AND AIMS: The prognostic relevance of sialyl Lewis X (sLeX) expression in colorectal and gastric cancer and its relevance to the hematogenous phase of tumor invasion is controversial. This study was designed to evaluate sLeX expression during tumor cell-endothelial cell interaction in vitro. METHODS: Adhesion and transendothelial penetration of MKN45, PaCa-2, WiDr, or Dan-G cells was analyzed by combined phase contrast-reflection interference contrast microscopy. In parallel, kinetics of membranous sLeX expression were examined fluorimetrically. To identify factor(s) which may be responsible for sLeX expression during tumor invasion tumor cells were treated with soluble immunomodulators, isolated endothelial plasma membranes, or E-selectin or P selectin IgG fusion proteins. sLeX was then analyzed by flow cytometry. RESULTS: Fluorometric quantification of sLeX demonstrated an inverse correlation between basal sLeX expression level and adhesion capacity of the tumor cells. Unexpectedly, sLeX was strongly down-regulated on tumor cell membranes in the course of heterophilic cell-cell contacts. The process occurred transiently, with a maximum effect 30-60 min after introducing tumor cells to the endothelial monolayer. Binding of tumor cells to immobilized E- and P-selectin IgG globulin chimeras was shown to be responsible for this phenomenon. CONCLUSION: A transient loss of sLeX is necessary for gastrointestinal tumor cells to invade endothelial cells. Due to the transient nature of the decrease in sLeX the controversy about the prognostic relevance of sLeX expression in colorectal and gastric cancer may be rooted in the stage of tumor invasion at the time of sLeX measurement. PMID- 12774244 TI - Angiogenesis and dendritic cell density are not correlated with metachronous distant metastasis in curatively operated rectal cancer. AB - BACKGROUND AND AIMS: Apart from surgery, treatment of rectal cancer increasingly involves the use of (neo-)adjuvant strategies. To optimize the selection process for these therapy regimens, especially in the field of cellular and molecular biology, new prognostic factors additional to the established TNM system are being investigated. PATIENTS AND METHODS: Two groups of patients ( n=2x85) with rectal carcinoma curatively treated by surgery alone were studied retrospectively (median follow-up 6.1 years). To exclude the effect of the surgeon only patients free of locally recurrent disease were selected. Patient groups were matched for age, gender, UICC stage, and year of operation (1982-1991) and differed only in subsequent metachronous distant metastatic spread, i.e., the criterion to be studied. The factors investigated in uni- and multivariate analysis were angiogenesis, density of dendritic cells, grading, venous invasion, and lymphatic invasion. RESULTS: Grading invariably proved to be the only significant prognostic factor. In univariate analysis the absence of venous invasion was also correlated significantly with increased disease-free survival. CONCLUSION: Angiogenesis and dendritic cell density are not prognostic factors for metachronous distant metastasis in rectal cancer and therefore cannot serve as selection parameters for adjuvant therapy. PMID- 12774245 TI - Reduction in alkaline sphingomyelinase in colorectal tumorigenesis is not related to the APC gene mutation. AB - BACKGROUND AND AIMS: The sphingomyelin pathway is an important intracellular mechanism in regulating cell growth. The first step in this pathway is catalysed by sphingomyelinases. Alkaline sphingomyelinase is specifically located in the intestinal tract. Markedly reduced alkaline sphingomyelinase activities have been found in sporadic colorectal tumours and in familial adenomatous polyposis adenomas. Since the adenomatous polyposis coli (APC) gene is mutated in about 80% of sporadic colorectal tumors, and familial adenomatous polyposis is the consequence of a germline mutation of the same gene, we examined whether low alkaline sphingomyelinase activity is linked to APC gene mutations. PATIENTS AND METHODS: Both germline and sporadic adenomatous polyposis coli gene mutations were studied. Alkaline, neutral, and acid sphingomyelinase activities were measured in the intestinal mucosa and content of multiple intestinal neoplasia mice, a murine model of familial adenomatous polyposis and compared to control mice. Alkaline sphingomyelinase activity was also measured in 11 human rectal tumors with APC gene mutation and compared with 9 control tumors without mutation. RESULTS: Alkaline, neutral, and acid sphingomyelinase activities were present in the small intestine and colon in both mice types with no differences in hydrolytic capacity or distribution pattern. In sporadic rectal tumors similar alkaline sphingomyelinase activities were identified in tumors with somatic APC gene mutations as in samples without mutations. In the tumors without detectable APC mutations beta-catenin was analyzed, but no mutation was detected. CONCLUSION: Alkaline sphingomyelinase is not directly linked to adenomatous polyposis coli gene mutations. PMID- 12774247 TI - Implications of reactive oxygen species and cytokines in gastroprotection against stress-induced gastric damage by nitric oxide releasing aspirin. AB - BACKGROUND AND AIMS: Nitric oxide-releasing aspirin (NO-ASA) has been shown to inhibit cyclo-oxygenase and prostaglandin generation without causing mucosal damage, but the role of reactive oxygen species (ROS) and cytokines in the action of ASA and NO-ASA against acute gastric damage has been little studied. METHODS AND MATERIALS: We compared the effect of NO-ASA and ASA on gastric lesions provoked by water-immersion and restraint stress (WRS), ischemia-reperfusion, and 100% ethanol. We determined the number and area of gastric lesions, gastric blood flow (GBF), plasma concentration of proinflammatory cytokines IL-1beta and TNFalpha, expression of superoxide dismutase (SOD) and glutathione peroxidase (GPx), ROS generation, and the malondialdehyde (MDA) concentration as an index of lipid peroxidation. RESULTS: Pretreatment with NO-ASA attenuated dose-dependently gastric erosions provoked by WRS, ischemia-reperfusion, and ethanol. In contrast, ASA aggravated significantly WRS-induced lesions, and this was accompanied by a fall in the GBF, suppression of prostaglandin E(2) generation, and significant rise in ROS chemiluminescence and in plasma TNFalpha and IL-1beta levels. ASA also enhanced significantly the mucosal MDA content and downregulated SOD and GPx mRNA, and these effects were markedly reduced by NO-ASA. CONCLUSION: Coupling of NO to ASA attenuates stress, ischemia-reperfusion, and ethanol-induced damage due to mucosal hyperemia mediated by NO, which compensates for prostaglandin deficiency induced by ASA. ASA aggravates WRS damage via enhancement of ROS and cytokine generation and suppression of SOD and GPx, and these effects are counteracted by NO released from NO-ASA. PMID- 12774246 TI - Pharmacological characteristics of the specific transporter for the endogenous cell growth inhibitor agmatine in six tumor cell lines. AB - BACKGROUND AND AIMS: This study examined agmatine transport into six human intestinal tumor cell lines and compared the pharmacological properties of this transporter with those of the agmatine carrier previously characterized in human glioblastoma cells. METHODS: Carrier-mediated uptake was determined as specific accumulation of [(14)C]agmatine in the cells. The changes in intracellular agmatine concentration in the tumor cells after 24 h incubation with 1 mM agmatine was analyzed by high-performance liquid chromatography. RESULTS: Specific [(14)C]agmatine accumulation was found in the six human intestinal tumor cell lines Caco2, Cx1, Colo320, HT29, Colo205E, and SW480. Specific [(14)C]agmatine accumulation was inhibited by phentolamine, putrescine, spermine, clonidine, and decynium-22 but not by corticosterone, O-methylisoprenaline, or l carnitine. Incubation with exogenous agmatine for 24 h increased intracellular agmatine content in all cell lines by a multiple of the basal endogenous content. Transfection of HEK293 cells with cDNA encoding either hOCT1, hOCT2, or hOCT3 did not enhance [(14)C]agmatine accumulation compared to nontransfected cells. CONCLUSION: All intestinal tumor cell lines investigated express a functional specific agmatine transporter which exhibit pharmacological characteristics similar to those of the agmatine transporter in glioblastoma cells. This agmatine carrier is not identical with any so far known organic cation transport system. PMID- 12774248 TI - A pilot study of irinotecan (CPT-11) as single-agent therapy in patients with locally advanced or metastatic esophageal carcinoma. AB - BACKGROUND AND AIMS: This study assessed the efficacy and safety of irinotecan (CPT-11) in the treatment of patients with unresectable esophageal carcinoma. PATIENTS AND METHODS: Ten patients with esophageal squamous cell carcinoma (SCC) and three with adenocarcinoma (AC) were treated with CPT-11. Eight of the 13 patients were pretreated by surgery, radio-, or chemotherapy. CPT-11 was administered in repeated 6-week cycles consisting of CPT-11 once weekly for 4 weeks, followed by a 2-week rest. The starting dose of CPT-11 was 125 mg/m(2) given intravenously over 60 min; subsequent doses were adjusted based on tolerance and toxicity. Nine patients were evaluable for response. RESULTS: Two patients showed a partial response (one SCC, one AC) and two others disease stabilization (one SCC, one AC). The mean time to progression was 3.8 months. Mean survival since study entry was 6.1 months. In the 103 administrations we observed grade 3 or 4 toxicity on six occasions with diarrhea, five with neutropenia, and one with nausea and vomiting. Toxicity required dose reductions in five patients; in two of these patients treatment was stopped because of severe toxicity. No treatment related deaths occurred. CONCLUSION: CPT-11 as single-agent therapy is modestly effective against squamous cell cancer of the esophagus. PMID- 12774249 TI - The pathological assessment of mesorectal excision: implications for further treatment and quality management. AB - BACKGROUND AND AIMS: Most clinical practice guidelines today recommend total mesorectal excision (TME) for carcinoma of the middle and lower rectal thirds and partial mesorectal excision (PME) for the upper rectal third. However, these procedures may not always fulfill the oncological requirements. The pathological examination of resected rectal carcinomas should always include a visual assessment of the mesorectal excision to ensure oncological adequacy and appropriate quality. The clinical practice guideline of the German Cancer Society recommends reporting of the distal extent of mesorectal excision (total or partial without coning) and the excision in an inviolate fascial envelope. PATIENTS AND METHODS: Reporting schemas of assessment and documentation for daily use and for studies are presented. RESULTS: Careful macroscopic evaluation of the resection specimen should be standardized. This may be supplemented by stain marking after postoperative filling the inferior mesenteric or superior rectal artery with ink or methylene blue solution. Photodocumentation is highly desirable. The pathological assessment of adequacy of mesorectal excision should be taken into account in selection for adjuvant radiotherapy. Objective macro- and microscopic assessment of mesorectal excision by pathologists is essential for quality management throughout patient care and in clinical trials. PMID- 12774250 TI - Rectocele repair improves evacuation and prolapse complaints independent of anorectal function and colonic transit time. AB - BACKGROUND AND AIMS: Evacuation disorders associated with a rectocele can be improved by rectocele repair. This study investigated whether anorectal function tests results change after rectocele repair. PATIENTS AND METHODS: Fourteen patients with 2nd or 3rd degree rectocele and evacuation disorder were treated by posterior colporrhaphy and evaluated pre- and postoperatively (after 8 months, range 3-14) using questionnaires, anal manometry and endosonography, rectal barostat testing, and colonic transit time measurement with radio-opaque markers. Results from female controls were used for comparison. RESULTS: Preoperatively, rectocele patients had high maximal basal sphincter pressures, large sphincter lengths, and low maximal squeeze pressures, with an anal sphincter defect in seven and lower visceral sensitivity scores than in controls. Postprandial rectal responses (more than 10% decrease in postprandial volume after 1 h) were found in 3 of 14 patients compared to 2 of 11 parous and 9 of 11 nulliparous controls. After repair, a rectocele of 2nd degree was found in four patients. Questionnaire scores were significantly decreased for straining, evacuation disorder, manual support, and protrusion. Overall patient satisfaction with the operation scored 8.25 (range 3-10). Defecation frequencies and stool consistencies were unaltered. Anal pressures, rectal compliance-curves, visceral sensitivity, and colonic transit times were unaltered after the rectocele repair. CONCLUSION: Rectocele repair improved complaints of evacuation disorder and protrusion, but defecation frequency and stool consistency were not affected. Anorectal function was unaltered after rectocele repair. Selection of patients for rectocele repair should be performed based on evacuation and protrusion complaints, anorectal function, or colonic transit time measurements have a limited role. PMID- 12774251 TI - Prospective study of artificial anal sphincter and dynamic graciloplasty for severe anal incontinence. AB - BACKGROUND AND AIMS: Dynamic graciloplasty and artificial anal sphincter are two options for refractory incontinence, the efficacy of which was compared in a prospective study. PATIENTS AND METHODS: Between November 1966 and June 1998, 16 patients were operated on (artificial anal sphincter 8, dynamic graciloplasty 8). Four consecutive operations with each technique were performed by two colorectal surgeons (one initiated the study with the neosphincter and the other with dynamic graciloplasty). Two independent observers assessed postoperative results at 4-month intervals. Patients were followed up to January 2001, with a median (interquartile range) of 44 (13) months and 39 (15) months for the nesophincter and the dynamic graciloplasty, respectively. RESULTS: Fourteen patients had complications. In the immediate postoperative period; there were eight cases of wound healing-related problems (four in the graciloplasty group). Perineal infection occurred in one patient in the graciloplasty group. At follow-up there were 11 complications (6 in the neosphincter group). Four patients undergoing neosphincter implantation had erosion or pain at the cuff site and had the implant removed (a new device was reimplanted in one). Four patients undergoing dynamic graciloplasty had the stimulator removed. Postoperatively the neosphincter was associated with a significantly lower score on the continence grading scale of the Cleveland Clinic Florida than graciloplasty. CONCLUSION: The artificial anal sphincter is a more convenient technique than dynamic graciloplasty for institutions treating small number of patients. However, technical failures and complications during follow-up that require reoperation are very high in both types of treatments. PMID- 12774252 TI - Current indications for blow-hole colostomy:ileostomy procedure. A single center experience. AB - BACKGROUND AND AIMS: Because of improved medical care and surgical techniques blow-hole colostomy with loop ileostomy is now rarely performed to reduce operative risks in patients with toxic megacolon related to inflammatory bowel disease (IBD). We reviewed patient charts to identify continuing indications for this procedure. PATIENTS AND METHODS: Seventeen patients underwent blow-hole colostomy procedure with ( n=15) or without ( n=2) ileostomy (8 men, 9 women; median age 51 years, range 21-79) during the past 18 years (1983-2001). RESULTS: The indications for the procedure were: toxic megacolon related to IBD ( n=6), toxic megacolon related to IBD and associated with pregnancy ( n=2), Clostridium difficile colitis ( n=3), adult Hirschsprung's disease ( n=1), pancreatitis with obstructing pseudocyst ( n=1), and palliation for malignant bowel obstruction with metastases ( n=4). Patients were discharged home after a median stay of 10 days (range 4-32 days). The 4 patients who underwent a palliative blow-hole procedure had died secondary to their underlying disease by the time of follow up. Of the remaining 13 patients 12 had their alimentary tract reconstituted, and one still awaits a definitive procedure. CONCLUSION: The blow-hole colostomy ileostomy procedure is still indicated for select patients with toxic megacolon and large-bowel obstruction. The procedure acts as a bridge to definitive operation for toxic patients with benign disease and palliates those with malignant obstructions and metastasis. PMID- 12774253 TI - Non-ligation of the hernial sac during herniotomy: a prospective study. AB - Herniotomy is performed for the surgical repair of hernia and along with orchiopexy for the closure of associated patent processus vaginalis. Ligation of the hernial sac has been considered mandatory for a successful repair. The present report was designed to study the results of non-ligation of the hernial sac before excision at the neck. It was found that non-ligation has no untoward effect on early complications and recurrence rate on long-term follow-up. It is suggested that it is not necessary to ligate the hernial sac during herniotomy in children. PMID- 12774286 TI - Racemization and degradation of thioridazine and thioridazine 2-sulfone in human plasma and aqueous solutions. AB - We present two methods for the enantioselective analysis of thioridazine (THD) and thioridazine 2-sulfone (THD 2-SO(2)) in human plasma based on liquid-liquid extraction with diethyl ether and chiral resolution of the enantiomers on Chiralpak AD and Chiralcel OD-H columns, respectively. After validation, the methods were used to study the degradation and racemization of both drug and metabolite. Our results showed that both enantiomers of THD and THD 2-SO(2) were stable at varying temperatures, pH, and ionic strengths; however, solubility problems for THD and THD 2-SO(2) enantiomers were observed, mainly at pH 8.5. The influence of light on the stability of the THD and THD 2-SO(2) enantiomers was also studied. Degradation of the THD enantiomers was observed under UV light (254 and 366 nm) while THD 2-SO(2) enantiomers were stable at these wavelengths and also when exposed to visible light. PMID- 12774287 TI - Preparation of enantiomeric gossypol by crystallization. AB - Large enantiomorphic crystals of gossypol-acetone (1:3) were grown from acetone solutions of rac-gossypol-acetic acid (1:1) at 4 degrees C. By controlling the initial gossypol concentration, crystallization time, and solution volume, single crystals were grown that weighed >50 mg, equivalent to >37 mg of enantiomeric gossypol. Even larger crystals were possible, but it was difficult to produce these reliably without contamination of the antipode. Essentially all of the acetone within the crystal form was removed by storing the crystals under vacuum for 3-4 days. By employing these techniques, gram quantities of enantiomeric gossypol were prepared in high chemical and optical purity. Based on measured and reported optical rotations, the optical purity of samples prepared by crystallization was greater than the optical purity of samples prepared by chromatographic separation of gossypol-amine diastereomers. The principal limitation of crystallization as a preparative method is the need to determine the chirality and purity of each product crystal. Nevertheless, the method competes favorably with preparative-scale chromatographic procedures. PMID- 12774288 TI - Enantiomeric separation by HPLC of 1,4-dihydropyridines with vancomycin as chiral selector. AB - The macrocyclic antibiotics represent a relatively new class of chiral selectors in CE, HPLC, and TLC. We have examined the use of the macrocyclic antibiotic vancomycin as a chiral selector in HPLC for the separation of 1,4 dihydropyridines (DHPs) calcium antagonists (CAs). Chromatographic data of six 1,4-dihydropyridine calcium channel blockers obtained on the vancomycin chiral stationary phase (Chirobiotic V) were compared with those obtained on an alpha(1) acid glycoprotein (AGP) HPLC stationary phase. Optimization of pH and organic modifier was carried out in order to modulate the retention properties of each system. All chiral neutral DHPs were resolved on the AGP column, whereas on Chirobiotic V only basic DHPs showed a split peak. The analytical chromatographic procedure on Chirobiotic V proved suitable for semipreparative separation, since the separation factor on the analytical column was high enough to obtain pure enantiomers with high yields. PMID- 12774289 TI - Effect of constituting amino acid residue numbers on molecularly imprinted chiral recognition sites. AB - In the present study, the effect of constituting amino acid residue numbers of oligopeptide derivatives, which are candidate materials to construct molecular recognition sites, on chiral recognition ability was investigated. Chiral recognition sites were formed from oligopeptide derivatives, of which constituting amino acid residue numbers were three to six, by adopting an alternative molecular imprinting. It was made clear that the number four, in other words, the tetrapeptide derivative, is the best candidate material to form a chiral recognition site. PMID- 12774291 TI - First asymmetric synthesis of (-)-lintetralin via intramolecular Friedel-Crafts type cyclization. AB - The asymmetric synthesis of an aryltetralin lignan, (-)-lintetralin, was achieved with an overall yield of 29% with seven steps. Key features of the synthesis are an asymmetric Strecker reaction, a diastereoselective Michael addition of the lithiated amino nitrile product to 5H-furan-2-one, and an intramolecular carbocationic cyclization to provide the desired ring skeleton with the correct configuration. PMID- 12774293 TI - Stereoselective metabolism of metoprolol: enantioselectivity of alpha hydroxymetoprolol in plasma and urine. AB - Direct stereoselective separation on chiral stationary phase was developed for HPLC analysis of the four stereoisomers of alpha-hydroxymetoprolol in human plasma and urine. Plasma samples were prepared using solid-phase extraction columns and urine samples were prepared by liquid-liquid extraction. The stereoisomers were separated on a Chiralpak AD column at 24 degrees C with fluorescence detection and a mobile phase consisting of a mixture of hexane:ethanol:isopropanol:diethylamine (88:10.2:1.8:0.2) for plasma samples and hexane:ethanol:diethylamine (88:12:0.2) for urine samples. Calibration curves for the individual stereoisomers were linear within the concentration range of 2.0 200 ng/ml plasma or 0.125-25 microg/ml urine. The methods were validated with intra- and interday variations less than 15%. The absolute configuration of the pure stereoisomers were assigned by circular dichroism spectra. The methods were employed to determine the concentrations of alpha-hydroxymetoprolol stereoisomers in a metabolism study of multiple-dose administration of racemic metoprolol to hypertensive patients phenotyped as extensive metabolizers of debrisoquine. We observed stereo-selectivity in the alpha-hydroxymetoprolol formation favoring the new 1'R chiral center from both metoprolol enantiomers (AUC(0-24) (1'R1'S) = 3.02). The similar renal clearances (Cl(R)) of the four stereoisomers demonstrated absence of stereoselectivity in their renal excretion. (-)-(S) metoprolol was slightly more alpha-hydroxylated than its antipode (AUC(0-24) (2S/2R) = 1.19), suggesting that this pathway is not responsible for plasma accumulation of this enantiomer in humans. PMID- 12774294 TI - Enantioseparation of racemic 4-aryl-3,4-dihydro-2(1H)-pyrimidones on chiral stationary phases based on 3,5-dimethylanilides of N-(4-alkylamino-3,5 dinitro)benzoyl L-alpha-amino acids. AB - Three novel chiral packing materials for high-performance liquid chromatography were prepared by covalently binding of (2S)-N-(3,5-dimethylphenyl)-2-[(4-chloro 3,5-dinitrophenyl)carbonylamino]propan-amide (7), (2S)-N-(3,5-dimethylphenyl)-2 [(4-chloro-3,5-dinitrophenyl)carbonylamino]-4-methylpentanamide (8), and (2S)-N (3,5-dimethylphenyl)-2-[(4-chloro-3,5-dinitrophenyl)carbonyl-amino]-2 phenylacetamide (9) to aminopropyl silica. The resulting chiral stationary phases (CSPs 1-3) proved effective for the resolution of racemic 4-aryl-3,4-dihydro 2(1H)-pyrimidone derivatives (TR 1-14). The mechanism of their enantioselection, supported by the elution order of (S)-TR 13 and (R)-TR 13 and molecular modeling of the complex of the slower running (S)-TR 13 with CSP 1 is discussed. PMID- 12774295 TI - Chiroptical properties and synthesis of enantiopure cis and trans pterocarpan skeleton. AB - The first enantioselective synthesis of trans-(6aS,11aR)-pterocarpan [(+)-2] and its conversion to cis-(6aS,11aS)-pterocarpan [(+)-1] was achieved starting from racemic 2'-benzyloxyflavanone (rac-3). Their stereochemistry was deduced by X-ray analysis of the ketal intermediate (-)-5a. The CD study of (+)-1 and (+)-2 allows the configurational assignment of similar pterocarpan derivatives by CD spectroscopy. PMID- 12774298 TI - Differential regulation by stimulants of neocortical expression of mrt1, arc, and homer1a mRNA in the rats treated with repeated methamphetamine. AB - The present work was conducted to obtain clues for the possible roles of a novel stimulant-inducible gene mrt1 (methamphetamine-responsive transcript 1) encoding a PDZ-PX protein in stimulant-induced behavioral sensitization. In the young adult rats, repeated daily treatment with methamphetamine (4 mg/kg, intraperitoneally, once a day) for 5 days caused an enhanced behavioral response to methamphetamine: behavioral sensitization. The 5-day intermittent administration of MAP upregulated the basal expression of mrt1 transcripts and eliminated the increasing effects of a challenge dose of MAP (1.6 mg/kg, i.p.) or cocaine (30 mg/kg, i.p.) on mrt1 expression on day 14 of withdrawal in the neocortex that has been considered to be composed of a neuron circuit implicated in the sensitization phenomenon. In contrast, the basal expression of other stimulant-inducible and plasticity-related genes arc and homer1a and the ability of MAP or cocaine challenge to augment the amounts of their transcripts were not affected by the repeated MAP regimen in the cortical area. These findings suggest the differential regulation by stimulant of neocortical mrt1, arc, and homer1a expression in the behaviorally sensitized animals and supports the view that stimulant induction of mrt1 may be involved in the early molecular signalings for stimulant sensitization. PMID- 12774299 TI - Higher in vivo muscarinic-2 receptor distribution volumes in aging subjects with an apolipoprotein E-epsilon4 allele. AB - The apolipoprotein E-epsilon4 allele confers an increased susceptibility to age related memory problems and Alzheimer's disease. Abnormalities in the cholinergic system are also likely contributors to memory deficits associated with aging and AD. To determine the effect of the APOE-epsilon4 allele on the muscarinic component of the cholinergic system of aging subjects, 10 healthy subjects with APOE-epsilon4 alleles (APOE-epsilon4+) and 10 without (APOE-epsilon4-), ranging in age from 52 to 75 years, were tomographically scanned with the F-18-labeled muscarinic-2 (M2) selective agonist, 3-(3-(3-[(18)F]Flouropropyl)thio)-1,2,5 thiadiazol-4-yl)-1,2,5,6-tetrahydro-1-methylpyridine ([(18)F]FP-TZTP). The distribution volumes (V(T)) of [(18)F]FP-TZTP were determined by compartmental modeling of partial volume and free fraction corrected PET scans. Regional cerebral blood flow (rCBF) measurements with H(2) (15)O were also performed. Global Gray V(T) (840 +/- 155 ml plasma/ml tissue) was greater in APOE-epsilon4+ subjects than APOE-epsilon4- subjects (660 +/- 113 ml plasma/ml tissue, P = 0.01), and previously studied younger subjects. There were no significant differences between the groups with respect to rCBF, but within the APOE epsilon4+ group there was a trend for subjects with the higher Global Gray V(T)s to have lower Global Gray CBFs (r = -0.65, P < 0.06). A lower concentration of acetylcholine in the synapse of APOE-epsilon4+ older individuals is a likely explanation for the greater [(18)F]FP-TZTP distribution volumes. PMID- 12774300 TI - Synaptic organization of monosynaptic connections from mesencephalic trigeminal nucleus neurons to hypoglossal motoneurons in the rat. AB - Synaptological characteristics of synapses between axonal boutons of the trigeminal mesencephalic nucleus (Vme) neurons and the hypoglossal nucleus (XII) motoneurons (MNs) were studied using biotinylated dextran amine (BDA) anterograde labeling combined with horseradish peroxidase (HRP) retrograde transport in the rat. BDA was initially iontophoresed into Vme unilaterally and 7 days later HRP was injected into the anterior two-thirds of the ipsilateral tongue. After histochemical reactions, BDA anterogradely labeled boutons were seen to appose closely to somata and dendrites of HRP retrogradely labeled MNs in XII by light microscopy. A total of 212 BDA-labeled Vme boutons were examined ultrastructurally, which had an average diameter of 1.3 +/- 0.4 microm and contain small clear spherical vesicles. Eighty-eight percent of Vme boutons (187/212) synapsed on dendrites of HRP-labeled XII MNs. Twenty-five Vme boutons (25/212, 12%) made synapses with somata of HRP-labeled XII MNs. Thirty-five percent (74/212) of BDA-labeled Vme boutons were also contacted by unlabeled P type terminals. Presynaptic P-type terminals contained spherical (47%, 35/74), pleomorphic (43%, 32/74), and flattened (10%, 7/74) synaptic vesicles. Thus, P type terminals (as a presynaptic element), BDA-labeled Vme boutons, and XII MNs constitute axoaxodendritic and axoaxosomatic synaptic triads. There are four types of synaptic microcircuits in XII neuropil: synaptic convergence, synaptic divergence, presynaptic inhibition synaptic circuits, and feedforward regulation circuits. This detailed ultrastructure examination of the synaptic organization between Vme neurons and XII MNs provides insights into the synaptic mechanisms of the trigeminal proprioceptive afferents involved in the jaw-tongue reflex and coordination during oral motor behaviors. PMID- 12774301 TI - [11C]-NS 4194 versus [11C]-DASB for PET imaging of serotonin transporters in living porcine brain. AB - In vitro, the novel diazabicyclononane NS 4194 has several thousand-fold selectivity for blocking the transport into rat brain synaptosomes of [(3)H] serotonin in comparison to [(3)H]-dopamine or [(3)H]-noradrenaline. We have prepared [(11)C]-NS 4194 in order to test its properties for PET imaging of brain serotonin transporters in comparison with the well-documented tracer [(11)C] DASB. Both compounds had rapid clearance from blood to brain of living pigs. The apparent equilibrium distribution volumes in cerebellum were 35 ml g(-1) for [(11)C]-NS 4194 and 11 ml g(-1) for [(11)C]-DASB. Pretreatment of pigs with citalopram did not reduce the uptake of either tracer in cerebellum, validating the use of that tissue as a nonbinding reference tissue for kinetic analysis of specific binding. The binding potential (pB) calculated for [(11)C]-NS 4194 using arterial input models was close to 0.5 in the telencephalon, and was 60% displaced by citalopram. However, the reference tissue method of Lammertsma was unsuited to calculate pB for this tracer, apparently due to its excessive nonspecific binding. In contrast to the relatively homogeneous binding of [(11)C] NS 4194, the pB of [(11)C]-DASB ranged from 0.6 in frontal cortex to 2 in the mesencephalon when calculated by the method of Lammertsma. Parametric maps of the pB of [(11)C]-DASB showed a pattern consistent with the known distribution of serotonin transporters in pig brain in vitro, and there was a uniform displacement of 80% of the specific binding after citalopram treatment in vivo. In conclusion, [(11)C]-DASB is in several respects superior to [(11)C]-NS 4194 for the detection of serotonin uptake sites by PET. PMID- 12774302 TI - Monoamine oxidase A imaging in peripheral organs in healthy human subjects. AB - Monoamine oxidase (MAO) catalyzes the oxidative deamination of many biogenic and dietary amines. Though studies of MAO have focused mainly on its regulatory role in the brain, MAO in peripheral organs also represents a vast mechanism for detoxifying vasoactive compounds as well as for terminating the action of physiologically active amines, which can cross the blood brain barrier. Indeed, robust central and peripheral MAO activity is a major requirement in the safe use of many CNS drugs, particularly antidepressants, and thus an awareness of the MAO inhibitory potential of drugs is essential in therapeutics. In this study, we examined the feasibility of quantifying MAO A in peripheral organs in healthy human subjects using comparative positron emission tomography (PET) imaging with carbon-11 (t(1/2): 20.4 min) labeled clorgyline ([(11)C]clorgyline) a suicide inactivator of MAO A and its deuterium labeled counterpart ([(11)C]clorgyline D2). Heart, lungs, kidneys, thyroid, and spleen showed a robust deuterium isotope effect characteristic of MAO and the magnitude of the effect was similar to that of trancylcypromine, an irreversible MAO inhibitor used in the treatment of depression. Liver time-activity curves were not affected by deuterium substitution precluding the estimation of liver MAO in vivo. In organs showing an isotope effect, MAO A is greatest in the lungs and kidneys followed by the thyroid and heart. This method, which has been previously applied in the human brain, opens the possibility to also directly assess the effects of different variables including smoking, dietary substances, drugs, disease, and genetics on peripheral MAO A in humans. PMID- 12774303 TI - Cytoskeletal changes in the hippocampus following restraint stress: role of serotonin and microtubules. AB - The aetiology of depression is associated with depletion in central levels of serotonin (5-HT). Hence, a major effect of antidepressant drugs is to increase synaptic 5-HT levels. Stressful conditions have also been shown to affect neuronal plasticity and 5-HT neurotransmission in the hippocampus. Neuronal plasticity, which is typically referred to as a structural adaptation of neurons to functional requirements, requires more dynamic forms of microtubules (cytoskeletal component). The alpha-tubulin, which is the major component of microtubules, can be postranslationally modified and both the tyrosinated (tyr tub) and acetylated (acet-tub) forms are considered markers of more dynamic or more stable microtubules, respectively. The aim of the present work was to investigate the expression of tyr-tub and acet-tub in the hippocampus of rats submitted to either acute (6 h for 1 day) or sub-chronic (6 h for 4 days every day) restraint stress. In addition, ex vivo hippocampal 5-HT levels were monitored by differential pulse voltammetry to analyse the influence of both stress conditions upon 5-HT levels. Our results showed that the expression of tyr tub in the hippocampus was significantly decreased to 70 +/- 7% following sub chronic restraint stress (P < 0.01). In contrast, acute and sub-chronic restraint stress increased the hippocampal expression of acet-tub to 139 +/- 11% and 145 +/ 11% of control, respectively. Finally, 5-HT levels were significantly increased (P < 0.05) to 142 +/- 15% and 135 +/- 11% following acute and sub-chronic restraint stress, respectively. The stress-induced cytoskeletal changes observed in the present study suggest that the microtubular network is a potential new pathway that may increase our understanding of stress-related events. PMID- 12774304 TI - Alpha 4 nicotinic acetylcholine receptor subunit links cholinergic to brainstem monoaminergic neurotransmission. AB - Agonists of nicotinic receptors containing the alpha4-subunit produce antinociception accompanied by several adverse side effects. The purpose of this study was to determine the distribution of the alpha4-subunit of nicotinic acetylcholine receptors (nAChR) in brainstem monoaminergic nuclei that may contribute to these effects using dual labeling immunofluorescence methods. The alpha4-subunit immunoreactivity was enriched in serotonergic (nucleus raphe magnus, pallidus, obscurus, and dorsalis) and noradrenergic (A5, locus coeruleus (LC), A7) areas associated with antinociception, where it was commonly colocalized with serotonin (5-HT) or tyrosine hydroxylase (TH) immunoreactivity. However, it was also noted that alpha4 was present in all other brainstem monoaminergic nuclei examined (adrenergic C1-C3, noradrenergic A1-alpha4, dopamine A9 and A10, nucleus raphe medianus). To determine if alpha4 agonists could impact neural activity in brainstem, monoaminergic nuclei that are associated with antinociception, the expression of c-Fos in response to the systemic administration of epibatidine (2.5, 5, or 10 microg/kg) was examined. Epibatidine produced a robust (2-5-fold) increase in c-Fos expression, which was not dose dependent, in all of these areas examined except the nucleus raphe magnus. These results suggest that the alpha4 subunit is positioned to mediate the effects of acetylcholine widely across many, if not all, monoaminergic neurons in the brainstem. These observations emphasize the potential involvement of noradrenergic, as well as serotonergic mechanisms in epibatidine's analgesic effects, and they also suggest that even selective alpha4 ligand may have widespread effects on brain monoamine neurotransmission. PMID- 12774305 TI - Stressor controllability modulates stress-induced serotonin but not dopamine efflux in the nucleus accumbens shell. AB - The ability of an organism to control a stressor can modulate many of the consequences of stress. Previous research indicates that uncontrollable stress (inescapable shock, IS) activates serotonergic (5-HT) neurons of the dorsal raphe nucleus (DRN) to a greater degree than controllable stress (escapable shock, ES), potentiating 5-HT efflux in the DRN and in target regions. Previous research also indicates that IS selectively affects dopamine (DA) efflux. The present study measured 5-HT and DA efflux in the NAc shell during IS or ES using in vivo microdialysis. Male Sprague Dawley rats with probes in the NAc shell were dialysed while subjected to 100 1.0 mA tailshocks. Rats were run in yoked pairs in wheel-turn boxes such that one rat (ES) in the pair could terminate the shock received by both himself and his yoked (IS) partner by a behavioral response. No stress controls remained in the dialysis bowls. 5-HT efflux selectively increased during IS, and remained increased throughout as well as after the stress session. There was no effect of stress on DA efflux. These results indicate that the NAc 5 HT response is preferentially sensitive to stress and can be modulated by stressor controllability. PMID- 12774306 TI - Grafted swine neuroepithelial stem cells can form myelinated axons and both efferent and afferent synapses with xenogeneic rat neurons. AB - Neuroepithelial stem cells derived from the swine mesencephalic neural tube were examined regarding their eligibility for neural xenografting as a donor material, with the aim of evaluating myelinated axon formation and both types of synaptic formation with xenogeneic host neurons as part of possible neural circuit reconstruction. The mesencephalic neural tube tissues were dissected out from swine embryos at embryonic days 17 and 18 and were implanted immediately into the striatum of the Parkinsonian model rat. The swine-derived grafts had many nestin positive rosette-forming, neurofilament-positive, and tyrosine hydroxylase positive cells in the rat striatum. Electron microscopic study revealed both efferent and afferent synaptic formations in the donor-derived immature neurons or tyrosine hydroxylase-positive donor cells in the grafts. Myelinated axons, both positive and negative for swine-specific neurofilament antibody, were mingled together in the graft. These results indicated that implanted neuroepithelial stem cells could survive well and divide asymmetrically into both nestin-expressing precursors and differentiated neurochemical marker-expressing neurons in the xenogeneic rat striatum, with the help of an immunosuppressant. Donor-derived immature neurons formed both efferent and afferent synapses with xenogeneic host neurons, and donor-derived axons were myelinated, which suggests that implanted swine neuroepithelial stem cells could possibly restore damaged neuronal circuitry in the diseased brain. PMID- 12774307 TI - Expression of depolarization-induced immediate early gene proteins in PC12 cells. AB - Immediate early genes induced by depolarization are thought to be important in mediating neuronal functional plasticity. We previously identified a group of immediate early genes that are preferentially induced by depolarization and forskolin but not by nerve growth factor or epidermal growth factor in PC12 pheochromocytoma cells. These depolarization-induced genes include synaptotagmin 4; the protein kinases KID-1, PIM-1, and SIK; an orphan transcription factor, Nurr-1; and a transcription corepressor, rTLE-3. All these genes are also induced in the hippocampus in response to kainic-acid induced depolarization. To characterize further the unique functions of these genes in plasticity, we used recombinant proteins to generate and purify antibodies against KID-1 and SIK proteins. Immunoblotting experiments were performed to examine the induced expression of the KID-1 and SIK proteins in PC12 cells. PIM-1 and Nurr-1 protein expression was also examined following stimulation, using commercially available antibodies. There is an increase in synthesis, in PC12 cells, of these four IEG proteins after KCl plus forskolin treatment. Nurr-1 protein peaks between 2 and 4 hr and decreases by 6 hr after the treatment. PIM-1 and KID-1 proteins rise by 1 hr, peak between 2 and 4 hr, and return to their basal levels at 6 hr. SIK protein increases significantly at 2 hr after treatment, peaks between 4 and 6 hr, and returns to the basal level at 8 hr. Immunofluorescence studies demonstrate distinct distribution patterns of each of these depolarization induced IEG proteins in PC12 cells. PMID- 12774308 TI - Oligodendrocyte progenitor migration in response to injury of glial monolayers requires the polysialic neural cell-adhesion molecule. AB - Injury to the nervous system results in reactive astrogliosis that is a critical determinant of neuronal regeneration. To analyze glial responses to mechanical injury and the role of the polysialic neural cell adhesion molecule (PSA-NCAM) in this process, we established primary glia cultures from newborn rat cerebral cortex. Scratching a confluent monolayer of primary glial cells resulted in two major events: rapid migration of oligodendrocyte progenitor-like (O-2A) cells into the wounded area and development of polarized morphology of type 1 astrocytes at the wound edge. Migrating O-2A progenitors had a bipolar morphology and exhibited A2B5 and O4 immunolabeling. Once these cells were established inside the wounded area, they lost A2B5 immunoreactivity and differentiated into glial fibrillary acidic protein-positive astrocytes. Migrating O-2A cells expressed PSA-NCAM, but type 1 astrocytes at the wound edge did not. Treatment of wounded cultures with Endo-N, which specifically removes PSA from the surface of cells, resulted in a significant decrease in O-2A cell migration into the wounded area and completely blocked the wound closure. Video time-lapse analysis showed that, in the presence of Endo-N, O-2A cells remained motile and migrated short distances but did not move away from the monolayer. These results demonstrate that O-2A progenitors contribute to reactive astrogliosis in culture and that PSA NCAM is involved in this process by regulating cell migration. PMID- 12774309 TI - Molecular mechanisms of glutamate neurotoxicity in mixed cultures of NT2-derived neurons and astrocytes: protective effects of coenzyme Q10. AB - Although glutamate excitotoxicity has long been implicated in neuronal cell death associated with a variety of neurological disorders, the molecular mechanisms underlying this process are not yet fully understood. In part, this is due to the lack of relevant experimental cell systems recapitulating the in vivo neuronal environment, mainly neuronal-glial interactions. To explore these mechanisms, we have analyzed the cytotoxic effects of glutamate on mixed cultures of NT2/N neurons and NT2/A astrocytes derived from human NT2/D1 cells. In these cultures, the neurons were resistant to glutamate alone (up to 2 mM for 24-48 hr), but they responded to a simultaneous exposure to 0.5 mM glutamate and 6 hr of hypoxia. Neuronal cell death occurred during subsequent periods of reoxygenation (>30% within 24 hr). This was associated with a marked decrease of intracellular ATP, a significant increase in reactive oxygen species (ROS) and downregulation of glutamate uptake by astrocytes. Thus, under energy failure and high levels of ROS production, only the neurons from these mixed cultures succumbed to glutamate neurotoxicity; the astrocytic cells remained unaffected by the treatment. Taken together, our data suggested that glutamate excitotoxicity might be due to the energy failure and oxidative stress affecting the properties of the NMDA glutamate receptors and causing impairment of glutamate transporters. Cells pretreated for 72 hr with 10 microg/ml of coenzyme Q(10) (functions both as a ROS scavenger and co-factor of mitochondrial electron transport), were protected, suggesting a useful role for coenzyme Q(10) in treatments of neurological diseases associated with glutamate excitotoxicity. A model of the complex interactions between neurons and astrocytes in regulating glutamate metabolism is presented. PMID- 12774310 TI - Transplantation of human adult astrocytes: efficiency and safety requirements for an autologous gene therapy. AB - Ex vivo gene therapy is emerging as a promising approach for the treatment of neurodegenerative diseases and central nervous system (CNS) trauma. We have shown previously that human adult astrocytes can be expanded in vitro and can express various therapeutic transgenes (Ridet et al. [1999] Hum. Gene Ther. 10:271-280; Serguera et al. [ 2001] Mol. Ther. 3:875-881). Here, we grafted normal and lentivirally-modified human adult astrocytes into the striatum and spinal cord of nude mice to test whether they are suitable candidates for ex vivo CNS gene therapy. Transplanted cells survived for at least 2 months (longest time analyzed) and sustained transgene expression. Importantly, the absence of proliferating cell nuclear antigen (PCNA) staining, a hallmark of cell division, ascertains the safety of these cells. Thus, adult human astrocytes are a promising tool for human CNS repair; they may make autologous ex vivo gene transfer feasible, thereby avoiding the problems of immunological rejection and the side effects of immunosuppressors. PMID- 12774311 TI - Testosterone up-regulates aquaporin-4 expression in cultured astrocytes. AB - Aquaporin-4 (AQP4) is located on astrocyte endfeet that face blood vessels in the brain and in the pia. It is thought to play a crucial role in the development of brain edema. To confirm the notion that sex steroids and dexamethasone influence brain edema through AQP4 regulation, we investigated the effects of 17beta estradiol, testosterone, and dexamethasone on the expression of AQP4 in cultured astrocytes. Testosterone significantly up-regulated AQP4 at the level of both protein and mRNA. At a concentration of 100 nM, testosterone significantly increased AQP4 protein levels and ameliorated the osmotic fragility of astrocytes from hypoosmotic stress, suggesting that the increased levels of AQP4 facilitated the testosterone function. Moreover, this effect was attenuated by the protein kinase C activator 12-O-tetradecanoylphorbol 13-acetate, which can rapidly decrease AQP4 mRNA expression, indicating that the response was specific. These results indicate that AQP4 can alter the osmotic fragility of astrocytes and that testosterone can influence brain edema through AQP4 regulation, whereas 17beta estradiol and dexamethasone cannot. PMID- 12774312 TI - Overexpression of V-1 prevents nitric oxide-induced cell death: involvement of enhanced tetrahydrobiopterin biosynthesis. AB - Previously we reported that the synthesis of catecholamines, dopamine, and noradrenaline was enhanced by overexpression of V-1 protein, a neuronal protein active in the initial stage of development of the rat cerebellum, in the neuronal cell line PC12D, a model of dopamine cells (Yamakuni et al. [1998] J. Biol. Chem. 273:27051-27054). To investigate the physiological role of this protein, we examined the effect of V-1 overexpression on cell toxicity induced by nitric oxide (NO) used at low concentrations. Two clones of PC12D cells overexpressing V 1, transfectants termed V1-46 and V1-69, were significantly more resistant to NOR3 (an NO donor) but not to etoposide (an inhibitor of topoisomerase II) induced apoptotic cell death than the control cells (termed C-7 and C-9) that had been transfected with the vector alone. The addition of L-DOPA, dopamine, or noradrenaline to the medium did not abolish NOR3-induced cell death in PC12D cells. Moreover, pretreatment of V1-46 and V1-69 cells with L-alpha-methyl-p tyrosine (alpha-MPT), an inhibitor of tyrosine hydroxylase, to inhibit catecholamine biosynthesis did not affect the resistance to NO toxicity. These results indicate that the catecholamine levels increased by V-1 overexpression did not produce the protection against NOR3-induced toxicity. We further showed that overexpression of V-1 enhanced the synthesis of (6R)-L-erythro-5,6,7,8 tetrahydrobiopterin (BH(4)). In addition, pretreatment with BH(4) or with sepiapterin, which is converted to BH(4) intracellularly, significantly protected PC12D cells in a dose-dependent manner. The increased BH(4) synthesis by V-1 overexpression was dose dependently inhibited by pretreatment with diaminohydroxypyrimidine (DAHP), an inhibitor of GTP-cyclohydrolase I, which is the rate-limiting enzyme for the biosynthesis of BH(4), concomitantly with the loss of protective effect afforded by V-1 overexpression. Furthermore, the addition of BH(4) or sepiapterin to DAHP-pretreated V146 and V1-69 cells restored cell viability. Taken together, these results indicate that V1 protein plays an important role in protection against cell death induced by NO at low levels by promoting the synthesis of BH(4). Moreover, these findings suggest the up regulation of V1 expression as a possible therapeutic target for protection against the insult of NO-induced oxidative stress. PMID- 12774313 TI - Lymphocyte infiltration in the injured brain: role of proinflammatory cytokines. AB - Studies using mouse axotomised facial motoneuron model show a strong and highly selective entry of CD3+ lymphocytes into the affected nucleus, with a maximum at Day 14, which coincides with the peak of neuronal cell death, microglial phagocytosis, and increased synthesis of interleukin-1 beta (IL1beta), tumour necrosis factor-alpha (TNFalpha) and interferon-gamma (IFNgamma). We explored the possible involvement of these cytokines during the main phase of lymphocyte recruitment into the axotomised facial motor nucleus 7-21 days after nerve cut using mice homozygously deficient for IL1 receptor type 1 (IL1R1-/-), TNF receptor type 1 (TNFR1-/-), type 2 (TNFR2-/-) and type 1 and 2 (TNFR1&2-/-), IFNgamma receptor type 1 (IFNgammaR1-/-), and the appropriate controls for the genetic background. Transgenic deletion of IL1R1 led to a 54% decrease and that of TNFR2 to a 44% reduction in the number of CD3+ T-cells in the axotomised facial motor nucleus, with a similar relative decrease at Day 7, 14, and 21. Deletion of TNFR1 or IFNgammaR1 had no significant effect. Deletion of both TNFR1 and 2 (TNFR1&2-/-) caused a somewhat stronger, 63% decrease than did TNFR2 deletion alone, but this could be due to an almost complete inhibition of neuronal cell death. No mutations seemed to inhibit aggregation of CD3+ T-cells around glial nodules consisting of Ca-ion binding adaptor protein-1 (IBA1)+ phagocytotic microglia and neuronal debris. Altogether, the current data show the importance of IL1R1 and TNFR2 as the key players during the main phase of lymphocyte recruitment to the damaged part of the central nervous system. PMID- 12774314 TI - Effect of tranexamic acid incorporated in fibrin sealant clots on the cell behavior of neuronal and nonneuronal cells. AB - Fibrin sealants are commonly used for hemostasis following surgery on various types of tissues. Aprotinin, an effective fibrinolysis inhibitor, is one of the components in some fibrin sealant products currently available. Tranexamic acid (tAMCHA) is another fibrinolysis inhibitor and is used as an alternative to aprotinin. Recent studies on fibrin sealant products containing tAMCHA indicate that it may be responsible for various adverse reactions when used in neurological applications. To determine a possible mechanism for such adverse reactions, we examined the effect of tAMCHA on the behavior of neuronal and nonneuronal cells using in vitro assays. The data indicate that different concentrations of tAMCHA incorporated in fibrin clots had no effect on the initial cell adhesion of either proliferative cells (glial cells and fibroblasts) or nonproliferative cells (neuronal cells) to the fibrin clots. Moreover, a high concentration of tAMCHA (300-450 mM) incorporated in the fibrin clots increased glial and fibroblast proliferation on fibrin clots. However, because tAMCHA is known to leach out of the fibrin clots, we have also examined the effect of solubilized tAMCHA in a growth medium on cells seeded on matrix-coated surfaces. A high concentration (300-450 mM) of tAMCHA detached all cell types from matrix coated dishes. Our model suggests that tAMCHA in fibrin clots has no adverse effect on cells bound to the fibrin clots; however, tAMCHA leaching out from the fibrin clots reduces adhesion of adjacent cells bound to their natural extracellular matrix. Thus, a high concentration of tAMCHA should not be used as a fibrinolysis inhibitor in fibrin sealant products, especially in neurosurgery. PMID- 12774316 TI - Transcription profiling reveals mitochondrial, ubiquitin and signaling systems abnormalities in postmortem brains from subjects with a history of alcohol abuse or dependence. AB - Alcohol abuse is a common human disorder with high rate of comorbidity with other psychiatric disorders. To identify candidate mechanisms for alcohol abuse, the expression of 12,626 genes was measured in postmortem temporal cortex from 11 subjects with a history of alcohol abuse or dependence, with or without other psychiatric diagnoses and compared pairwise with the expression in 11 nonalcoholic subjects matched for the other psychiatric diagnoses and demographics. Genes were defined to have altered expression in alcohol abuse if: 1) the gene showed decreased expression in at least 10 of 11 subjects with alcohol abuse, or showed increased expression in at least 10 of 11 subjects with this diagnosis compared to matched non-abusers (P < 0.007, chi(2)test); or 2) the difference in the mean abuser/non-abuser ratio for the gene from value of 1.0 was significant at P < 0.05 (one sample t-test). In subjects with a history of alcohol abuse or dependence, 163 genes were changed significantly. The most abundant and consistent changes were in gene families encoding mitochondrial proteins, the ubiquitin system, and signal transduction. These alterations indicate disturbances in energy metabolism and multiple signaling mechanisms in the temporal cortex of subjects with a history of alcohol abuse or dependence. We hypothesize that these mechanisms may be related to alcohol abuse traits or long term effects of alcohol. PMID- 12774315 TI - Effects of peroxisome proliferator-activated receptor gamma ligands ciglitazone and 15-deoxy-delta 12,14-prostaglandin J2 on rat cultured cerebellar granule neuronal viability. AB - Peroxisome proliferator-activated receptor gamma (PPARgamma) has been the focus of studies assessing its potential neuroprotective role. These studies have shown either neuroprotection or neurotoxicity by PPARgamma ligands. Comparison of these studies is complicated by the use of different PPARgamma ligands, mechanisms of neurotoxicity induction, and neuronal cell type. In this study, we compared the effects of the synthetic PPARgamma ligand ciglitazone with an endogenous PPARgamma ligand, 15-deoxy-delta(12,14)-prostaglandin J(2) (15-deoxy PGJ(2)), on inherent neurotoxicity and neuroprotection using a reduction in extracellular KCl in rat cultured cerebellar granule neurons (CGN). We also assessed the effects of these ligands on c-Jun protein expression, which is up-regulated on induction of low-KCl-mediated neuronal apoptosis as well as being associated with PPAR in other cell types. We showed that PPARgamma mRNA is expressed in CGN cultures and observed ciglitazone- and 15-deoxy PGJ(2)-mediated inherent neurotoxicity that was concentration and time dependent. c-Jun was only modestly increased in the presence of ciglitazone but was markedly up-regulated by 15-deoxy PGJ(2) after 12 hr. Treatment of CGN cultures with ciglitazone simultaneous with KCl withdrawal resulted in a modest, time-dependent neuroprotection. Such neuroprotection after KCl withdrawal was not observed with 15-deoxy PGJ(2). Despite the absence of neuroprotection, 15-deoxy PGJ(2) markedly inhibited the early up-regulation of c Jun during KCl withdrawal. These studies suggest that ciglitazone and 15-deoxy PGJ(2) have markedly different effects on inherent and low-KCl-induced toxicity and c-Jun expression in CGN, indicating potential non-PPARgamma mechanisms. PMID- 12774317 TI - Cyclic AMP responsive element binding protein phosphorylation and persistent expression of levodopa-induced response alterations in unilateral nigrostriatal 6 OHDA lesioned rats. AB - Activation of cAMP responsive element binding protein (CREB) has been increasingly implicated in the formation and maintenance of long-term memory. To elucidate molecular mechanisms that underlie the persisting alterations in motor response occurring with levodopa (L-dopa) treatment of parkinsonian patients, we evaluated the time course of these changes in relation to the activation of striatal CREB in 6-hydroxydopamine (6-OHDA) lesioned animals. Three weeks of twice-daily L-dopa treatment reduced the duration of the rotational response to acute L-dopa challenge in hemiparkinsonian rats, which lasted about 5 weeks after withdrawal of chronic L-dopa therapy. This shortened response duration, resembling human wearing-off fluctuations, was associated with a marked increase in Ser-133 phosphorylated CREB (pCREB) immunoreactivity in medium spiny neurons in dorsolateral striatum in response to acute dopaminomimetic challenge. Intermittent treatment with the D1 receptor-preferring agonist SKF 38393, but not the D2 receptor-preferring agonist quinpirole, produced a similar rise in CREB phosphorylation. The time course of changes in CREB phosphorylation correlated with the time course of changes in motor behavior after cessation of chronic L dopa therapy. Both the altered motor response duration and the degree of CREB phosphorylation were attenuated by the intrastriatal administration of CREB antisense or protein kinase A inhibitor Rp-cAMPS. The results suggest that region specific Ser-133 CREB phosphorylation in D1 receptor containing spiny neurons contributes to the persistence of the motor response alterations produced by intermittent stimulation of striatal dopaminergic receptors. PMID- 12774318 TI - Enhanced binding of nor-binaltorphimine to kappa-opioid receptors in rats dependent on butorphanol. AB - Autoradiographic characterization of binding for brain kappa(1) ([(3)H]CI-977) and kappa(2) ([(3)H]bremazocine) in the presence of DAMGO ([D-Ala(2),N-Me Phe(4),Gly(5)-ol]-enkephalin), DPDPE ([D-Pen(2), D-Pen(5)]-enkephalin), and U 69,593 opioid receptors, in the presence of different concentrations of a selective unlabeled kappa-opioid receptor antagonist, nor-binaltorphimine (nor BNI), was performed in rats in which dependence on or withdrawal from butorphanol had been established. Dependence was induced by a 72 hr intracerebroventricular (i.c.v.) infusion with butorphanol (26 nmol/microl/hr; butorphanol dependent). Butorphanol withdrawal was produced by terminating the infusion of butorphanol in dependent animals. Responses were studied 7 hr following termination (butorphanol withdrawal). IC(50) values from competition studies were estimated by fitting inhibition curves for both kappa(1)- and kappa(2)-opioid receptor assays. In both dependent and withdrawal groups, the IC(50) values obtained against [(3)H]CI-977 or [(3)H]bremazocine with nor-BNI were decreased (ratios of approximately 0.03 0.21 and approximately 0.05-0.42 vs. control, respectively) in brain regions, including frontal cortex, nucleus accumbens, claustrum, dorsal endopiriform nucleus, caudate putamen, parietal cortex, posterior basolateral amygdaloid nucleus, dorsomedial hypothalamus, hippocampus, posterior paraventricular thalamic nucleus, periaqueductal gray, substantia nigra, superficial gray layer of the superior colliculus, ventral tegmental area, and locus coeruleus, compared with control. These results indicate that, in butorphanol-dependent and butorphanol-withdrawal rats, the brain kappa(1)- and kappa(2)-opioid receptors developed a supersensitivity to antagonist binding. PMID- 12774319 TI - [Effects of the active components of some Chinese herbs on drug metabolizing enzymes]. AB - OBJECTIVE: To review the effects of the active components of Chinese herbs on drug metabolizing-enzymes. METHOD: Relevant research papers reported in recent years were consulted and studied. RESULT: The drug metabolizing-enzymes cytochrome P450 and UDP-glucuronosyl transferase and glutathione S-transferase were inhibited or induced by the flavonoids, furocoumarins, and the active components extracted from salvia miltiorrhiza and hypericum perforatum, and so on, which therefore slowed or sped metabolism of other drugs in vivo and in vitro. CONCLUSION: Much attention should be paid to the metabolic interaction of the Chinese herbs when coadministered with other drugs. PMID- 12774320 TI - [Studies on the pollen morphology of ten species of Achillea]. AB - OBJECTIVE: To study the pollen morphology of ten species of Achillea. METHOD: The pollen morphology of ten species of Achillea was examined with LM and SEM. RESULT: The pollen grains were usually 3-colporate, subspheroidal. The exine ornamentation consisted of verrucate, spinulate and foveolate. But some differences in size, colpae and exine ornamentation were found. CONCLUSION: The slight differences of the pollen morphology are useful to some extent for the classification of the ten species of Achillea. PMID- 12774321 TI - [Determination and contrastion of alkaloids and saponins in Bulbus Fritillariae Cirrhosae and in other beimus]. AB - OBJECTIVE: To establish quality standard of Bulbus Fritillariae Cirrhosae by determining and comparing alkaloids and saponins in various Beimus. METHOD: Alkaloids in beimus were determined with two-phase titration and saponins in Beimus were determined with weight method. RESULT AND CONCLUSION: The contents of alkaloids and saponins in 4 species of Bulbus Fritillariae Cirrhosae (each specie including 3 samples) Zhe Beimu, Yi Beimu and Pin Beimu (each specie including 2 samples) were determined. PMID- 12774322 TI - [Determination of piceid in commercial Rhizoma Polygoni Cuspidati by HPLC]. AB - OBJECTIVE: To improve a RP-HPLC method of determining piceid in commercial Rhizoma Polygoni Cuspidati. METHOD: HPLC method was used, with C18 column (250 mm x 4.6 mm), a mixture of CH3CN-H2O (20:80) as the mobile phase, and detecting wavelength 303 nm. RESULT: The linear range was 0.0066-0.0792 microgram.microL-1, r = 0.9999 (n = 5). The average recovery rate was 98.5% and RSD was 2.5% (n = 9). CONCLUSION: The method is simple, accurate and reproducible. The contents of piceid in commercial Rhizoma Polygoni Cuspidati from different areas are variable and the average content is 2.23%. PMID- 12774324 TI - [Study on stem-tip tissue culture of the traditional Chinese medicine Chrysanthemum morifolium]. AB - OBJECTIVE: To set up the optimums for the stem-tip tissue culture of Chrysanthemum morifolium cultivated in Anhui Province. METHOD: Small sections (about 0.5 mm in length) from the stem-tips were isolated and inoculated with different media, and induced to form the whole plantlet formation. RESULT AND CONCLUSION: The MS medium added with 6-BA 2 mg.L-1 + NAA 0.2 mg.L-1 was the optimum medium for the bud sprouting and the inducing rate was over 80% after 40 d cultivation on this modified medium. The MS medium supplemented with 6-BA 2 mg.L-1 and NAA 0.5 mg.L-1 was the optimum medium for the multiplication of the adventitious buds in which the bud multiplication was about 4-7 times higher after 25-30 d cultivation. The plantlet could root well on the MS medium with NAA 0.5 mg.L-1. PMID- 12774323 TI - [HPLC determination of 6-gingerol in Rhizoma Zingiberis Recens]. AB - OBJECTIVE: To determine the contents of 6-Gingerol in Rhizoma Zingiberis Recens. METHOD: HPLC method was used, with Alltech C18 column, acetonitrile-methol-water (43:5:52) as mobile phase with a flow rate of 0.8 mL.min-1, detecting wavelength 280 nm, and column temperature 35 degrees C. RESULT: Retained time of 6-gingerol was near 19 min, showing a good recovery (98.2%) and linear correlation (r = 0.9999). The contents of 6-gingerol were 1.35-2.87 mg.g-1, and the water contents were 70.4-85.5% mL.g-1 in Rhizoma Zingiberis Recens. CONCLUSION: The method is appropriate for the determination of 6-gingerol in Rhizoma Zingiberis Recens. Gingerol can be used as a chemical marker of the quality control of Rhizoma Zingiberis Recens. PMID- 12774325 TI - [Preparation of genistein-loaded chitosan microspheres]. AB - OBJECTIVE: To optimize the preparation of genistein chitosan microspheres with central composite design (CCD). METHOD: The chitosan microspheres were prepared by the O/W/O multiple emulsion method. Second-order polynomial and linear equations were fitted to the data, and the resulting equations were used to produce response surface graphs and the best experiment conditions. RESULT: The theoretical drug content was 13%-15%, the concentration of organic phase was 30% 40% and the concentration of oil phase was 68%-72%. CONCLUSION: The best experiment conditions can be obtained by central composite design and response surface methodology. The observed values agree well with model predicted values. PMID- 12774326 TI - [Fingerprint analyses of fructus Forsythiae]. AB - OBJECTIVE: To provide application basis of the Forsythia suspensa by studying the difference of HPLC-FP of F. suspensa fructification (medicinal materials). METHOD: Comparative work was done on F. suspensa produced in different areas, on different parts of Forsythia suspensa, and on the pseudo preducts with methods of HPLC-FP. RESULT: Different FP characteristics were shown respectively by different samples, which were from different producing areas, from different parts, and the pseudo products including the fructification of Syringa reticulata var. and F. viridissimac. CONCLUSION: The FP can be used to distinguish the F. suspensa coming from different producing areas and different sources. PMID- 12774327 TI - [A zanhic acid based bisglycoside from Aster poliothamnus]. AB - OBJECTIVE: To study the chemical constituents from the whole plant of Aster poliothamnus. METHOD: Separating the chemical constituents by means of chromatography and identifying ther structures on basis of chemical and spectral tecnology. RESULT AND CONCLUSION: A new triterpene saponin was isolated and identified. PMID- 12774328 TI - [The changes of some chemical components in banxiaxiexintang decoction of different combinations]. AB - OBJECTIVE: The contents of bererine, palmatine and glycyrrhizin acid in Banxiaxiexintang decoction of different combinations were determined by PR-HPLC. METHOD: A Shim-pack CLC-ODS column was used with a mobile phase of CH3CN-H2O (31:69; 0.005 moL.L-1 -pentanesulfonic acid sodium salt, H3PO4: pH 3.0) for bererine andpalmatine, which were detected at the wavelength of 275 nm. A YWG-C18 column was used with a mobile phase of CH3OH-H2O-HAc(62:37:1) for glycyrrhizin acid which was detected at the wavelength of 260 nm. RESULT: Each herbs' combination influences the contents of the 3 components. CONCLUSION: The experiment is an attempt to study the comical foundation of traditional Chinese prescription. PMID- 12774329 TI - [The effect of shourong compound formula on levels of dopamine and its metabolites in brain of Parkinson's disease mice induced by reserpine]. AB - OBJECTIVE: To investigate the effect of Shourong compound formula on treating Parkinson's disease. METHOD: Parkinson's disease model mice induced by reserpine was used and by HPLC-ED the levels of Dopamine (DA) and its metabolites were determined. RESULT: Madopar could increase the levels of DA in brain of PD mice. The effect of madopar together with Sourong compound formula was better than that of madopar(P < 0.001). CONCLUSION: Shourong compound formula together with madopar has synergic effect on increase of DA level in brain and can reduce clinic dose of madopar so that side effect of madopar can be decreased. PMID- 12774330 TI - [Protective effects and its mechanism of panaxatriol saponins isolated from Panax notoginseng on cerebral ischemia]. AB - OBJECTIVE: To study the protective effects and its mechanism of Panaxatriol Saponins isolated from Panax notoginseng (PTS) on focal cerebral ischemia in rat brain. METHOD: The influences of PTS on cerebral water content and three specific proteins (VEGF, HSP70 and transferrin) related with cerebral ischemia were studied with unilateral occlusion of the middle cerebral artery (MCAO) and Western Blot. RESULT: PTS 12.5 mg.kg-1 i.p. x 7 d (5 d before MCAO and 2 d after MCAO) inhibited the increase of cerebral water content caused by MCAO and influenced contents of HSP70 and transferrin, but had no influence on VEGF protein level. CONCLUSION: PTS shows a protective effect on focal cerebral ischemia in rat brain by alleviating cerebral edema, up-regulating the expression of HSP70, down-regulating transferrin and maintaining blood-brain barrier. PMID- 12774331 TI - [Effect of protein and anthraquinone glucosides from cassia seed on serum lipid of hyperlipidemia rats]. AB - OBJECTIVE: To study the effect of protein and anthraquinone glucosides from cassia seed on serum lipid of hyperlipidemia rats. METHOD: The rat hyperlipidemia model was set up by ig lipid emulsion. The effects of the protein 0.25 and 1 mg.kg-1. d-1, the anthraquinone glucosides 5 and 20 mg.kg-1.d-1, and the protein 0.25 mg.kg-1.d-1 plus the anthracene glucosides 5 mg.kg-1.d-1 on total cholesterol (TC), triglyceride (TG), low density lipoprotein (LDL-C) and high density lipoprotein (HDL-C) in serum of the rats were determined. RESULT: The protein 1 mg.kg-1.d-1, the anthraquinone glucosides 20 mg.kg-1.d-1 reduced the raised TC, TG, LDL-C of hyperlipidemia rats (P < 0.05). The above indexes could also be reduced by the protein 0.25 mg.kg-1.d-1 plus the anthraquinone glucosides 5 mg.kg-1.d-1(P < 0.05, P < 0.01). CONCLUSION: Protein and anthraquinone glucosides from cassia seed can lower TC, TG, LDL-C of hyperlipidemia rats. PMID- 12774332 TI - Calcium prevents loss of glutathione and reduces oxidative stress upon reperfusion in the perfused liver. AB - BACKGROUND: Addition of 1.5 mM Ca2+ to the preservation solution (UW) during static cold rat liver preservation have been shown to improve liver function upon reperfusion. Effects of adding calcium to the perfusate during liver machine perfusion are yet not described. METHODS: A recently developed model for rat liver machine perfusion (hypothermic oscillating oxygenated liver perfusion) was used to perfuse rat livers with calcium free modified UW solution (Group 1) or with modified UW + 2 mM CaCl2 (Group 2) for a period of 10 h (4 degrees C). In both experimental groups an acellular reperfusion at 37 degrees C with Ringer solution and 50 microM ferricytochrome c over a period of 90 min was performed. Liver and perfusate samples were taken before and after reperfusion to assess the cellular energy charge, metabolites, parameters of oxidative stress, cellular calcium, bile flow, enzyme release and TNF alpha. RESULTS: Hypothermic perfusion with oxygenated calcium free modified UW solution resulted in depletion of cellular calcium and glutathione. Upon reperfusion bile function was inhibited in spite of a sufficiently reloaded energy charge and low LDH release. In contrast machine perfusion with modified UW solution +2 mM Ca2+ prevented the loss of both, cellular calcium and glutathione during the preservation period and led to sufficient bile flow and less release of superoxide anions upon reperfusion. CONCLUSIONS: The loss of cellular calcium and glutathione during oxygenated machine liver perfusion appeared to be reducible by adding 2 mM Ca2+. Furthermore, upon reperfusion, livers with preserved cellular calcium demonstrated significantly lower oxidative stress and an improved liver function. PMID- 12774333 TI - S-adenosylmethionine immunomodulator treatment in sepsis. AB - BACKGROUND: S-adenosylmethionine (SAMe) is a forerunner of glutathione. AIMS: The aim of the present study is to ascertain this drug effect on T-lymphocytes and cytokines in an experimental model of surgical sepsis. METHODS: Rats were allotted in two groups. In the control group, rats underwent anaesthesia and laparatomy with cecal ligation and puncture (CLP). In the second group, rats underwent the same CLP and received SAMe (14 mg/kg) i.m., on the 1st (1PO) and 2nd (2PO) postoperative days. A week before surgery (PRE), on the 1PO and on the 3PO: IL-1, IL-2, IL-4, IL-6, IL-10 and TNF levels (ELISA & MoAb), and CD3, CD4, CD8 cell and IL-2R percentages (%) (flow cytometry & MoAb) were determined in peripheral blood. RESULTS: Rats receiving SAMe do not show changes of CD3%, CD4% and IL-1 levels but show a significant increase of CD8% on the 3PO, showing a significant difference with regard to controls (p < 0.01). Both groups show a similar IL-2R variation pattern: increasing on 1PO (p < 0.05) and decreasing on 3PO (p < 0.05). CONCLUSION: In sepsis: SAMe inhibits the decrease of circulating immune-active cells and the IL-1 increase. This drug seems to have effects useful in avoiding immunological alterations in sepsis, that need to be tested in humans. PMID- 12774334 TI - A comparison of the microvascular response in the healing wound in the spontaneously hypertensive and non-hypertensive rat. AB - BACKGROUND: In the Spontaneously Hypertensive Rat (SHR), there is a significantly greater blood flow at the paw but not at the back than in the non-hypertensive Wistar Kyoto (WKY) rat. We wanted to assess the effect of this higher blood flow on wound healing at the paw. MATERIALS AND METHODS: We characterized the microvascular composition of wounds at the back and paw of 9 SHR rats and 10 WKY rats using a quantitative imaging program. Blood flow was compared using laser Doppler technology. RESULTS: The blood flow response to wounding at the back was identical in the SHR and WKY rats. There was an immediate sharp increase in flow at the center of the wound. Blood flow reached a peak at 3 days and then decreased somewhat by day 7, but still remained five-fold higher than the prewound baseline values. There was also a two-fold increase at the back wound perimeter. There were no differences in microvascular composition at the back between the SHR and WKY rats. In contrast, there was an immediate enormous increase in blood flow at paw wound center in the SHR rats. Flow increased to 75 ml/min/100 gm by 24 h then fell back sharply. Blood flow at the paw in the WKY rats changed very little over the 7 days post wounding. At 3 days, the flow was about twice as high in the SHR than in the WKY wound, but, by day 7, flow was similar in the two rat strains. At the SHR wound perimeter, there was a small increase in flow which was sustained through day 7. Although the microvascular composition at the paw wound center was similar in the SHR and WKY rats, there was a notable difference at the paw perimeter. At baseline, there was a slightly greater capillary density in the SHR paw (32 +/- 1 per mm3) than the WKY paw (25 +/- 8 per mm3). At 7 days after wounding, there was a substantial increase in capillary number in the SHR rats (48 +/- 8 per mm3) as compared to baseline (p = 0.05). In contrast, there was no significant difference in capillary number in the WKY paw wound perimeter (20 +/- 3 per mm3) as compared to baseline. CONCLUSIONS: There is a substantial difference in wound blood flow response between the hypertensive and the non-hypertensive rat. At the back, the blood flow effects of wounding are similar, but, at the paw, the SHR rat shows a dramatic transient increase in flow in the early phases of wound healing. There is apparently no capability to upmodulate microvascular resistance in response to increased pressure at this early stage of wound healing. However, within several days, the granulation tissue microvasculature becomes capable of controlling the effects of raised pressure in the SHR rat. In the SHR paw wound perimeter, there are significantly more capillaries than in the WKY rat. It is possible that greater capillary proliferation in the SHR rat results from higher blood flow in the early phase of wounding. The contrast between the WKY rat and the SHR rat serves to further illustrate the complexity of blood flow regulation which occurs during wound healing. PMID- 12774335 TI - Pylorus preservation after complete gastrectomy and jejunal interposition: experimental study investigating motility and alkaline reflux. AB - BACKGROUND: Pylorus preservation has been proposed to reduce the disturbance in gastric emptying following gastrectomy but little is known about the role of the pylorus regarding motility patterns and alkaline reflux. AIMS: To assess the motility patterns and alkaline reflux following pylorus preservation after complete gastrectomy and jejunal interposition in the pig. METHODS: Motility patterns and alkaline reflux were studied in 12 conscious pigs before and after gastrectomy and jejunal interposition with preservation of the pylorus and a 1 cm antral ring (PYL+) or excised pylorus (PYL-) 6 and 14 weeks postoperatively. Gastroduodenal motility was investigated by manometry after fasting and fed, respectively and alkaline reflux by a fiberoptic system for detecting bilirubin. RESULTS: The number of propagated phase-III activities of the jejunal interponat to the duodenum and propagating velocity were lower in the PYL+ group after 6 weeks (p < 0.05) but higher after 14 weeks (p < 0.05) compared to the PYL- group. In the PYL+ the number of intrapyloric pressure waves (IPPWs) was lower after 6 weeks (p < 0.05) but higher after 14 weeks (p < 0.05) in comparison to preoperative controls. Alkaline reflux was significantly higher in the PYL- group than in the PYL+ group throughout the time observed. CONCLUSIONS: Pyloric function shows a delay after 6 weeks but restores within 14 weeks postoperatively. Pylorus preservation was associated with a significant decrease of alkaline reflux compared to PYL- in this setting. PMID- 12774336 TI - Short-term symptomatic outcome and quality of life after laparoscopic versus open Nissen fundoplication: a prospective randomized trial. AB - BACKGROUND: Laparoscopic operation has replaced conventional operation in the treatment of reflux disease. This change has been mostly based on excellent results from highly experienced antireflux surgeons rather than on randomized clinical trials. AIMS: The objective of this study was to compare the short-term symptomatic outcome and patient quality of life costs after laparoscopic (LNF) or open Nissen fundoplication (ONF) in a community hospital setting with less experienced surgeons. METHODS: Forty-two patients with documented gastroesophageal reflux disease (GERD) were randomized to either LNF or ONF. Symptomatic outcome using a custom questionnaire and the Gastrointestinal Quality of Life Index (GIQLI) were measured pre- and postoperatively at one and three months. RESULTS: Esophagitis was cured among all patients in LNF group compared to 90% in the ONF group. The symptoms observed preoperatively were significantly improved in both groups, except for dysphagia and flatulence. Dysphagia was more common after LNF. The GIQLI (scale 0-144) was equally normalized in both groups. The mean GIQLI-change among all patients was 37.9 points. Patient satisfaction did not differ between the groups. CONCLUSIONS: LNF and ONF are effective methods in the operative treatment of GERD in short-term and result in a significant improvement in patients gastrointestinal symptoms and quality of life. PMID- 12774337 TI - Thromboembolism prophylaxis and incidence of thromboembolic complications after laparoscopic surgery. AB - The aim of this prospective study was to assess the clinical thrombo-embolic risk in laparoscopic digestive surgery. METHODS: The study prospectively included 2384 patients, who underwent laparoscopic surgery between June 1992 and June 1997. All patients received peri-operative low molecular weight heparin (LMWH) thromboprophylaxis. This regimen was administered until the patient resumed normal ambulatory activity. RESULTS: Eight cases (0.33%) of deep vein thrombosis (DVT) were observed, but no pulmonary embolism was noted. In 6 cases (5 cholecystectomies with reverse Trendelenburg position and 1 inguinal hernia repair), the pneumoperitoneum was more than 2 h, and in 2 cases (1 rectopexy and 1 sigmoid colectomy for diverticulitis), more than 3 h. In 6 out of the 8 cases, the diagnosis of DVT was established after cessation of LMWH delivery, after the patients were discharged home, and before post-operative day 10. CONCLUSION: During laparoscopic surgery, long operations and reverse Trendelenburg position are potentiating factors to DVT. Heparin prophylaxis for laparoscopic procedures should continue at least until discharge, and continued prophylaxis after discharge should only be considered in individual patients at continued high risk. We also recommend using graduated compression stockings, maintaining a relatively low insufflation pressure, keeping use of the reverse Trendelenburg position to a minimum, and intermittently releasing the pneumoperitoneum in longer procedures. PMID- 12774338 TI - Cytokeratin 20 is not a tissue-specific marker for the detection of malignant epithelial cells in the blood of colorectal cancer patients. AB - BACKGROUND: The clinical benefit of using reverse transcription polymerase chain reaction (RT-PCR)-based assays to detect circulating tumour cells during post operative surveillance of cancer patients remains unclear. Cytokeratin 20 has been proposed as a tissue-specific marker for the detection of micrometastases in the blood of colorectal cancer patients. However, recent reports have challenged its specificity, and hence the validity of its use. AIMS: The aim of this study was to evaluate the tissue-specificity of ck20 mRNA transcription and its use for detecting circulating colon epithelial cells. PATIENTS AND METHODS: RNA was isolated from the peripheral blood of 51 colorectal cancer patients, four patients with benign gastrointestinal disease and 42 healthy controls. In addition, it was prepared from 32 colorectal cancers, from a pituitary cancer, from normal kidney, liver, fibroblasts, keratinocytes and from 24 lymph nodes obtained from eight patients with benign gastrointestinal disease. Real-time RT PCR assays were used to quantitative and compare ck20 transcription. RESULTS: Significant levels of ck20 mRNA were detected in all 42 blood samples from healthy volunteers and in all pre- and post-operative blood samples from colorectal cancer patients regardless of the presence of metastatic disease. It was also detected in all other mRNA samples analysed. CONCLUSION: The lack of colon tissue-specificity renders ck20 useless as a marker for the post-operative surveillance of colorectal cancer patients. PMID- 12774339 TI - Panniculectomy in morbidly obese gynecologic oncology patients. AB - BACKGROUND: It is clear that morbid obesity presents a serious risk to women undergoing major intra-abdominal gynecologic surgery. Unfortunately, many gynecologic malignancies and benign conditions are best treated surgically. Thus, the gynecologic surgeon must choose an incision that permits adequate exposure with acceptables rates of complications. Panniculectomy as an approach to the peritoneal cavity is becoming increasingly popular. Recent reports suggest that panniculectomy is reasonably straightforward to perform, provides adequate exposure and is associated with an acceptable rate of manageable complications. In these circumstances, panniculectomy is not a cosmetic procedure, but medically necessary to perform indicated major gynecologic intra-abdominal surgery. AIMS: To present our experience with panniculectomy in morbidly obese women undergoing major intra-abdominal surgery on a gynecologic oncology service. METHODS: The medical records of 48 morbidly obese women (Quetelet Index > 40 kg/m2) with a large dependent pannus who underwent major intra-abdominal surgery via a panniculectomy between May 1990 and October 1999 were reviewed. Data regarding demographics, concomitant medical conditions, operative indications and results, and postoperative outcomes were abstracted for analysis. RESULTS: The mean age was 54.9 years, the mean body mass was 130.2 kg, the mean height was 1.63 m and the mean Quetelet Index was 49.3 kg/m2. The mean operating time was 188 min and the mean estimated blood loss was 615 ml. Two patients suffered intraoperative urologic injuries which were repaired without sequelae; there were no bowel, vascular or neurologic injuries. Eighteen patients had a suprafascial wound breakdown (3 complete and 15 superficial) and 15 patients developed an infection. There were no documented deep venous thromboses, pulmonary emboli or fascial dehiscences. Two patients died in the postoperative period; one from a myocardial infarction on Day 2 and one from overwhelming sepsis on Day 76. CONCLUSIONS: Morbid obesity is associated with substantial operative and postoperative risks. Panniculectomy provides operative exposure with acceptable risks of complications. PMID- 12774340 TI - Tumor development and cytokine production by human colon tissues and carcinoma cell lines. AB - BACKGROUND AND OBJECTIVES: Solid tumors evade the host immunologic responses they initiate by unknown mechanisms. The authors investigated patterns of cytokine content in human colon carcinomas, colon cancer cell lines in vitro, and nude mouse xenografts from those lines in order to clarify those mechanisms. METHODS: Epithelial tumor cell lines were developed from specimens of human colon adenocarcinoma. Aliquots of these cells were then xenografted into female heterozygous BALB/c nu/+ immunologically deficient mice and serially passaged. Original tumors, cell lines, and resultant xenografts were then analyzed for histology/cytology and for levels of TGF-beta and TNF-alpha by enzyme linked immunoassay. RESULTS: Cytokine levels were elevated beyond baseline mucosal levels in original tumors and xenograft mouse tumors but not detectable in extracts from epithelial cultures. CONCLUSIONS: While the precise source of cytokine production remains unclear, these data suggest tumor/host interactions not found in pure epithelial cancer cells in culture. PMID- 12774341 TI - Role of DIC in multiple organ failure. AB - BACKGROUND: Disseminated intravascular coagulation (DIC) is a severe and complex coagulopathy resulting from excessive thrombin formation. Although there is a final common pathway, consumption coagulopathy and multiple organ failure (MOF), the early pathophysiology differs depending on the underlying disease. AIM: The aim of this study is to elucidate the pathophysiology of septic DIC, and also to assess logical diagnostic and therapeutic procedure. METHODS: Blood samples were collected from septic patients. Coagulant, fibrinolytic and vascular markers were analyzed. RESULTS AND CONCLUSION: Damaged vascular endothelial cells overproduce plasminogen activator inhibitor 1, which, with excess production of thrombin, leads to fibrnolysis-suppressive DIC and fibrin formation in the microvasculature. These changes characterize the septic coagulopathy, which is often complicated by MOF; however, a bleeding tendency is relatively rare. The platelet count and fibrinogen/fibrin degradation products are most useful for the diagnosis of septic DIC. However, since the specificity is not sufficient with these screening tests, other tests including molecular markers should be added. Since endothelial damage plays an important role in the pathophysiology in septic DIC, treatment should be focused not only on abnormal coagulopathy but also on vascular damage. Protease inhibitors and antithrombin III are both effective treatments for DIC. Although heparin is a standard drug for the treatment of DIC, the use of heparin is not recommended in case of septic DIC. PMID- 12774342 TI - Congenital absence of the portal vein in liver transplantation for biliary atresia. AB - BACKGROUND: Extrahepatic biliary atresia is the most common indication for liver transplantation in children. Coexistent congenital anomalies are common (25%), but the combination of polysplenia syndrome (10%) and absence of portal vein is rare. AIMS: We report a case of successful reduced size liver transplantation on a 13-month-old girl with extrahepatic biliary atresia, polysplenia syndrome and congenital absence of the portal vein. METHODS: Technical and post-operative problems related to absence of the portal vein and polysplenia are described and the literature reviewed. RESULTS: The donor portal vein was successfully anastomosed to the confluence of the superior mesenteric vein and splenic vein. The child is well with good graft function at a follow-up of 20 months. CONCLUSION: Absence of the portal vein and polysplenia syndrome is not a contraindication for liver transplantation although an increased post-operative morbidity may be expected. PMID- 12774343 TI - Telomerase and breast cancer: from diagnosis to therapy. AB - BACKGROUND: Telomeres are believed to be responsible for chromosomal stability during cellular proliferation. They are created by telomerase, an enzyme present in many human neoplasms. Telomerase is considered to be important in breast cancer since reactivation of the enzyme ensures cell stability allowing malignant cells to divide indefinitely. AIMS: This article aims to discuss the role of telomerase in breast cancer and explore the future of telomerase research. METHOD: We have reported on the literature to date in order to present this current review. RESULTS: Telomerase is found in significant levels in breast neoplasms and is reliably detected in fine needle aspirations aiding the diagnosis of breast cancer. There is also a correlation between telomerase and disease prognosis, and there is current work assessing if telomerase can be of benefit in monitoring the efficacy of systemic treatment. CONCLUSION: The role of telomerase in breast cancer is still being investigated however by understanding the action it has on chromosomal stability, and its detection in tumour cells, there are possibilities to enhance the methods employed to detect and treat breast cancer. PMID- 12774344 TI - [Study on complex system of Chinese materia medica GAP fulfilling]. AB - OBJECTIVE: To provide new model of thinking and management for Chinese Meteria Medica GAP fulfilling. METHOD: After analyzing the problems of GAP fulfilling, and considering GAP a complex system, we had some initial discussions about the systematic structure, characteristics and the methods for fulfilling it. RESULT: The GAP was a complex system which consisted of biological system, environment system and management system with its own characteristics and methods. CONCLUSION: The GAP fulfilling still has some problems and needs the direction of systematic opinion of complex system. PMID- 12774345 TI - [Probe into characteristics of Taraxacum mongolicum ultramicro-power]. AB - OBJECTIVE: To observe and study the tissue characteristics of T. mongolicum ultramicro-power and dissolving-out characteristics of effective compositions. METHOD: By microscopic observation and thin-layer chromatography. RESULT: Nearly all cell walls of T. mongolicum are broken and dissolving-out characteristics of effective compositions are remarkably improved, after it is ultramicro porphyrized. PMID- 12774346 TI - [The characters of various commodity of chrysanthemums]. AB - OBJECTIVE: To provide basis for making the quality standard of medicinal chrysanthemums [Dendranthema morifolium (Ramat.) Tzvel.]. METHOD: The character of pharmacognosy. RESULT: The characters of many of chrysanthemums as well as the differences between them have been clarified. PMID- 12774347 TI - [Study on the physicochemical properties of cultivated soil of genuine crude and no-enuine crude Chinese Angelica]. AB - OBJECTIVE: To provide basis for environmental factors of genuine crude Chinese angelica. METHOD: On-the-spot investigation and indoor chemical analysis were made to study the physicochemical properties of cultivated soil of Chinese angelica. RESULT AND CONCLUSION: The physical properties and the organic matte and mineral nutrition of cultivated soil of Chinese angelica are best in Min County, Gansu Province. The ecological environment is the leading factor in forming genuine crude Chinese angelica. PMID- 12774348 TI - [Determination of cirsilineol in Herba Artemisiae scopariae collected in autumn by HPLC]. AB - OBJECTIVE: To establish a quantitative method for determination of cirsilineol in Herba Artemisiae Scopariae collected in Autumn. METHOD: Sample was extracted with methanol, and cleaned up with polyamide column. ODS column was used with methanol acetonitrile-0.5% acetic acid solution (24:19:57) as mobile phase. Detection wavelength was 347 nm. RESULT: Cirsilineol in sample solution was well separated. Linearity of cirsilineol was good (r = 0.9998) in range of 0.1-0.8 microgram. The average recovery was 101.3%, RSD of repeatability was 3.04%. CONCLUSION: This method can be used for quality control of Herba Artemisiae Scopariae collected in autumn and its preparations. PMID- 12774349 TI - [Determination of lignan glycosides in Gaultheria leucocarpa var. yunnanensis by RP-HPLC]. AB - OBJECTIVE: To control the quality of Gaultheria leucocarpa var. yunnanensis the contents of lignans in these plants from different provinces were detected. METHOD: HPLC was adopted using Hypersil-18 column (0.4 cm x 25 cm), methanol: acetonitrile: water (30:5:65) as mobile phase. RESULT: The calibration curves of D1(gaultherinA), D2[(-)-5'-methoxyisolariciresinol-2 alpha-O-beta-D xylopyranoside)], D4[(+)-lyoniresinol-2 alpha-O-beta-D-glucopyranoside)] were linear in the range of 0.10-0.40 microgram, 0.03-0.13 microgram, 0.02-0.09 microgram, respectively. The average recovery of D1 was 98.9% and RSD was 2.6%. CONCLUSION: Comparing the samples from Jinxiou(158.2 mg.100 g)-1 and Kumming(154.8 mg.100 g)-1, that of Guiyang showed the highest content of LD(208.8 mg.100 g)-1. The content of LD in the tender above-ground part was 1.3 times higher than that in the wilt. PMID- 12774350 TI - [Comparative studies on isolation and purification of Chinese medicine decoction]. AB - OBJECTIVE: To evaluate different methods by treating water-decocted liquid of 6 Chinese medical herbs and 4 co-prescription respectively with alcohol, ultrafilter, macroporousresin and clarifier. METHOD: The contents of target component in those extracts were determined with HPLC or titration, and quantitative and qualitative determination of the impurity components, such as polysaccharide and protein, was made. RESULT: Each method showed its advantages and disavantages. CONCLUSION: Different method can be chosen according to the clinical and preparation demands or the characteristic of components. PMID- 12774351 TI - [Studies on chemical constituents of effective part of Gastrodia elata]. AB - OBJECTIVE: To study the chemical constituents of effective part of Gastrodia elata. METHOD: The constituens were isolated by column chromatographies with silica gel and ODS, and identified by NMR, MS spectra. RESULT: Six compounds were isolated and identified. CONCLUSION: Three compounds were isolated from this plant for the first time. PMID- 12774352 TI - [Two flavone glycosides from Chinese traditional medicine Amomum villosum]. AB - OBJECTIVE: To study the water soluble constituents from Amomum villosum. METHOD: The constituents were separated and purified with chromatographic methods, identified by NMR, MS, UV and IR. RESULT: Two quercetin glycosides: quercitrin (quercetin-3-O-alpha-L-rhamnoside I) and isoquercitrin (quercetin-3-O-beta-D glucoside II) were isolated and identified. CONCLUSION: I and II were isolated for the first time from A. villosum. PMID- 12774353 TI - [Studies on the phenylpropanoids from Eucommia ulmoides]. AB - OBJECTIVE: To study the chemical constituents from the leaves of Eucommia ulmoides. METHOD: The constituents were isolated by chromatography method and the structures were identified on the basis of spectral analysis. RESULT: Six compounds, ursolic acid(1), beta-sitosterol(2), p-coumaric(3), caffeic acid ethyl ester(4), chlorogenic acid(5) and syringin(6) were obtained. CONCLUSION: Compound 3, 4, 5 were obtained from the plant for the first time. PMID- 12774354 TI - [Lipid compounds from Echinacea purpurea]. AB - OBJECTIVE: To study the lipid constituents from Echinacea purpurea. METHOD: The compounds were isolated by chromatography method and the structures were identified on the basis of spectral analyses. RESULT: Five compounds were isolated and identified as, 1 beta, 6 alpha-dihydroxy-4(14)-eudesmene(1), (2E, 4E, 8Z, 10E)-N-isobutyl-2,4,8,10-dodecatetraenamide(2), (2E, 4E, 8Z, 10Z)-N isobutyl-2,4,8,10-dodecatetraenamide(3), cerotic acid(4), hyxacosyl alcohol(5). CONCLUSION: Compounds 1,4 and 5 were obtained from the plant for the first time. PMID- 12774355 TI - [Studies on chemical constituent of Solanum lyratum]. AB - OBJECTIVE: To study chemical constituents of Solanum lyratum. METHOD: The constituens were isolated by polystyrene resin RA, sephadex LH-20 and silica gel column chormatography, their structures were identified by spectroscopic analysis. RESULT: Four compounds were identified as The constituens were caffeic acid, vanillic acid, rutin and N(p-hydropheneethyl) p-coumaramide. CONCLUSION: These compounds were isolated from this plant for the first time. PMID- 12774356 TI - [Studies on the chemical constituents in the leaves of Cyclocarya paliurus]. AB - OBJECTIVE: To investigate the chemical components of the leaves of Cyclocarya paliurus. METHOD: The chemical components were isolated by solvent extraction and column chromatography. The chemical structures were identified on the basis of physic-chemical constant and spectral data. RESULT: Six compounds were isolated and identified as kaempferol(I), quercetin(II), isoquercitrin(III), ellagic acid(IV), daucosterol(V) and cyclocaric acid B(VI). CONCLUSION: Compounds I, II, III and IV were obtained for the first time from this plant. PMID- 12774357 TI - [Reversal of anti-apoptotic action by tetrandrine in human breast carcinoma multidrug-resistant MCF-7 cells]. AB - OBJECTIVE: To study whether the anti-apoptotic action is reversed by tetrandrine in a combination with vincristine in human breast carcinoma MCF-7 multidrug resistant cells. METHOD: Chromatin condensation was observed by co-staining of fluorescent dyes Hoechst 33342 and propidium iodide; and G1 sub-peak was detected by flow cytometry. Apoptotic cells were detected with TUNEL method. Cellular free ca2+ was determined with Fluo-3 staining method. RESULT: Two types of chromatin condensation were observed after the sensitive and drug-resistant MCF-7 cells were treated with an antitumor drug vincristine 5 mumol.L-1 for 24 h. The number of cell with chromatin condensation was obviously reduced in the drug-resistant cells treated with the same concentration of vincristine, as compared with the sensitive MCF-7 cells. The number of the apoptotic cells was increased by a combination of non-cytotoxic tetrandrine 20 mumol.L-1 and vincristine in both the sensitive and drug-resistant cells, which was confirmed with fluorescent indication and TUNEL method. The increment of introcellular free Ca2+ level in the cells treated with tetrandrine in a combination of vincristine was detected with Fluo-3 staining method. CONCLUSION: The anti-apoptotic action of human breast carcinoma MCF-7 cells can be effectively reversed by tetrandrine. PMID- 12774358 TI - [Effect of components of dang-gui-bu-xue decoction on hematopenia]. AB - OBJECTIVE: To investigate the effects and the related mechanisms of the components of Dang-Gui-Bu-Xue decoction (DGBXD) on improving blood deficiency. METHOD: The effects of promoting hematopoietic function were observed with the blood difficient model mice, by giving components of DGBXD. RBC, WBC, reticulocytes and bone marrow nucleated cells (BMNC) were determined. The components of DGBXD on proliferation of BMNC and on clony forming unit (CFU) were also determined. RESULT: The components of DGBXD remarkably increased the quantity of RBC, WBC, and BMNC. Some of the components promoted the proliferation of BMNC and increased the quantity of CFU-Mix. Among them, polysaccharide of angelica was most potent. CONCLUSION: The studies show that the extracts and some components of DGBXD can promote the hemopoietic function system of the model mice, and they exert the effects in a comprehensive way. PMID- 12774359 TI - [Screening of antimitotic portions from sea hare using conidia of Pyricularia Oryzae]. AB - OBJECTIVE: To apply conidia of Pyricularia Oryzae to the screening of antimitotic constituents from marine animal sea hare. METHOD: To extract and fractionate active portions from sea hare through detecting deformation of mycelia germinated from conidia of P. Oryzae P-2b, in comparison with the cytotoxic test results in vitro. RESULT: Two active portions, of which IC50 against P388 and HL-60 was 23.4, 18.6 and 19.4, 12.5 micrograms.ml-1, respectively, were screened from this animal. CONCLUSION: This bioassay method was efficiently applied to the primary screening of antimitotic portions from marine animals for the first time. Being convenient, speedy and cheap, the screening model is suitable for the bioassay of active constituents from marine life. PMID- 12774360 TI - [Study on the quantitative change of anthraquinonoids of Rhei in the preparation of dachengqi]. AB - OBJECTIVE: To study the scientific evidence of the traditional preparation of Dachengqi: "Boiling Aurantii Immaturus and Magnoliae Officinalis first, and then adding Rhei to decoct together. Discarding the dregs, adding Natrii Sulfas into the decoction and drinking the upper solution when the Natrii Sulfas has dissolved completely". METHOD: The concentrations of free and combined anthraquinonoids(emodin, rhein, chrysophanol, physcion) in different decoctions were determined with HPLC method respectively. RESULT: When Natrii Sulfas, Aurantii Immaturus and Magnolias Officinalis are decocted with Rhei in different schemes, the concentrations of anthraquinonoids were changed regularly. CONCLUSION: The scientific evidence of traditional preparation method greatly increased the concentrations of the active components in Dachengqi. PMID- 12774361 TI - [Opinion of colon-targeting delivery about rhubarb extract as a purgative]. AB - OBJECTIVE: To elucidate that rhubarb extract as a purgative should be delivered to colon from a pharmaceutical point of view. METHOD: Effects of anthranoids and free anthraquinones as purgatives on the colon motility and transit time, the absorption in vivo, and the loss in the processes of extraction, concentration and dryness were reviewed, based on the recent 60 years pharmacological, pharmaceutical and clinical experimental studies at home and abroad. RESULT AND CONCLUSION: Free anthraquinones in rhubarb extract should be regarded as purgative principles. Close attention should be paid to the loss of free anthraquinones not only in the processes of extraction, concentration and dryness, but also in the process of transit in the upper gastrointestinal tract. Colon-targeting delivery of rhubarb extract, as a purgative, may prevent absorption of free anthraquinones in the upper gastrointestinal tract, thus improving clinical effects and lowering dosage. PMID- 12774362 TI - [Summarization of the clinical and laboratory study on the rhubarb in treating chronic renal failure]. AB - OBJECTIVE: To review the clinical and laboratory study on the rhubarb in the treatment of Chronic Renal Failure (CRF). METHOD: Documents of clinical and mechanism study on chinese medicine rhubarb including the simple rhubarb and the rhubarb complex prescription in which rhubarb is the main effective ingredient in recent 10 years were searched and summarized. RESULT: Clinical application involving dosage forms, prescriptions, usage and effect of the rhubarb and its prescriptions in the treatment of the CRF and their laboratory study progress wre summarized. CONCLUSION: Rhubarb and all kinds of its complex prescriptions have an assured effect on CRF and their mechanism includes many ways such as ameliorating azotemia, preventing nephritic compensatory hypertrophy and high metabolism situation, and so on. So the rhubarb can be used as an ideal medicine in treating CRF in a long period of time and will have a very brilliant prospect. PMID- 12774363 TI - [The influences of inorganic elements in soil on the geolism of Atractylodes lancea]. AB - OBJECTIVE: To study the influence of inorganic elements in soil on the geolism of Atractylodes lancea. METHOD: The concentrations of 15 kinds of inorganic elements of regional A. lancea and their correspondingly soils were determined, and the data were analysed with the SPSS10.0 software. RESULT: The mean concentration of Ni in geoherbs of A. lancea was 3 times higher than that in the non-geoherbs. The accumulation of Ca is high in geoberbs. There is no relationship between the concentrations of inorganic elements in A. lancea and those in their corresponding soil. CONCLUSION: A. lancea adjusts the concentration of inorganic elements mainly by active absorption, and there are good relationship between the accumulation of Fe and Cr, Fe and Hg, Hg and Cr, As and Pb, Co and Sr. PMID- 12774364 TI - [Identification of sea cucumber sold in market]. AB - OBJECTIVE: To identify the variety of the sea cucumber sold in market. METHOD: Figure and bone pieces observed for the microstructures. RESULT: 8 varieties were identified as Lessonothurea Deichmann, Thymiosycia Pearson, Microthele Brandt and Metriatyla Rowe of Holothuria, Bohadschia, Stichopus and Thelenota. CONCLUSION: The bone pieces in ora, anus, back, abdomen and tentacle of sea cucumber are different. They are the important bases to identify sea cucumber. PMID- 12774365 TI - [Comparison study on total flavonoid content and anti-free redical activity of the leaves of bamboo, phyllostachys nigra, and Ginkgo bilabo]. AB - OBJECTIVE: To investigate the differences of total flavonoid (TF) content and antifree radical activity between the-leaves of bamboo and Gingo biloba, as well as their seasonal changes. METHOD: Spectrophotometery and Chemiluminescence methods were adopted to determine TF and half inhibiting concentration (IC50) on active oxygen free radicals of the leaves of bamboo, phyllostachys nigra (Lodd. ex. Lindl.) Munro, and Ginkgo biloba. Two kinds of leaves were picked in the same plot at the same time monthly. RESULT: The TF of bamboo leaf varied in the range of 0.67%-1.71% (in dry basis of leaf, below as same) throughout a year, the minimum apparing in June and the maximum in July, then going down obviously, and remaining at a much high lever during November to next April. However, the TF of Ginkgo bilabo leaf varied in 1.48%-2.49% during whole growing period, early April to late November. It ascended with the growth of leaf, reaching the top during June and July, the going down slowly, and finally another peak appeared before defoliation. The average IC50 values on O2-. and .OH of bamboo leaf were at 11.0 micrograms.mL-1 and 5.3 mg.mL-1, and Ginkgo biloba at 19.0 micrograms.mL-1 and 3.6 mg.mL-1, respectively. CONCLUSION: The TF content and anti-free radical activity the bamboo leaf are comparable with the leaf of ginkgo biloba, which is a kind of potential resources for natural antioxidant and free radical scavenger. PMID- 12774366 TI - [Measurement of baicalin contents by HPLC]. AB - OBJECTIVE: To compare the contents of baicalin in the raw medical material and the different prepared medical materials of Scutellaria baicalensis. METHOD: The contents of baicalin were determined by HPLC. Chromatographic conditions included Hypersil C18 column and the mobile phase consisting of a mixture of methanol 0.04% phosphoric acid (46:54). Baicalin was detected at 280 nm. The standard curve was linear in the range of 0.0280-4.6700 micrograms with correlation coefficient 1.0000. The average recovery of baicalin was 101.22% with RSD = 2.54% (n = 5). RESULT: The contents of baicalin were 6.80% in raw material, 6.00% and 6.73% in the different prepared medical materials of S. baicalensis respectively. CONCLUSION: The two different kinds of preparing methods can be used to the raw material of S. baicalensis. PMID- 12774367 TI - [Determination of verbascoside in herba of Galeobdolon Chinense by RP-HPLC]. AB - OBJECTIVE: To determine Verbascoside in Galeobdolon chinense. METHOD: HPLC method was used, with the column packed with Kromasil C18(4.6 mm x 150 mm, 5 microns). The mobile phase was a mixture of acetonitrile and 1% acetic acid in a ratio of 13:87, and the ultraviolet wavelength was set at 350 nm. RESULT: The average recovery was 97.0%, the RSD being 1.69%. CONCLUSION: This method is sensitive and reliable. It can be used to determine being Verbascoside in Galeobdolon chinense. PMID- 12774368 TI - [Chemical components of essential oils from Flos chrysanthemi Indici in different areas]. AB - OBJECTIVE: To provide the foundation for reasonable utilization by analysing the essential oils of Flos chrysanthemi Indici in different areas. METHOD: The essential oils were extracted by using steam distillation and separated with GC capillary columns. The components were quantitatively determined with normalization method, and were identified with GC-MS. RESULTS: 18, 17 and 20 compounds of essential oils from Guangxi, Guangdong and Hubei were identified. CONCLUSION: There are significant differences among the components and contents of essential oils of Flos chrysanthemi Indici from Guangxi, Guangdong and Hubei. PMID- 12774369 TI - [Qualitative and quantitative methods of chlorogenic acid in commercial Herba Artemisiae Scopariae]. AB - OBJECTIVE: To develop the identification and assay for chlorogenic acid in commercial Herba Artemisiae Scopariae. METHOD: TLC method was used for identification with silica gel G plate and butyl acetate-formic acid-water (7:2.5:2.5) upper layer as a developing solvent. Chlorogenic acid in 85% methanolic extract was separated on the ODS column with methonal-3% acetic acid solution (15:85) as mobile phase. The detection wavelength was 327 nm. RESULT: The qualitative method is repeatable. Chlorogenic acid in extracts is well separated, relationship of injection amount and peak area is linear (r = 0.9998) within the range of 0.075-0.6 microgram. The average recovery is 100.9% and repeatability is 1.4%. Ten samples purchased from different areas in the country were identified and quantified with the methods. CONCLUSION: The methods and data could be used for quality control. PMID- 12774370 TI - [Studies on the chemical constituents from marine brown alga Ishige okamurai (3)]. AB - OBJECTIVE: To study the chemical constituents of Ishige okamurai. METHOD: Compounds were isolated by Pyricularia oryzae bioassay-guided fractionation method in combination with extraction and partitionation as well as multi chromatography. Their structures were determined by spectral analysis and chemical evidence. RESULT: Seven compounds were obtained and identified as (2S)-1 O-palmitoyl-2-O-(11Z-octadecenoyl)-3-O-beta-D-galacto-pyranosyl glycerol (I), (2S)-1-O-palmitoyl-2-O-myristoyl-3-O-(6-sulfo-alpha-D- quinovopyranosyl) glycerol (II), (2S)-1-O-palmitoyl-2-O-palmitoyl-3-O-(6'-sulfo-alpha-D- quinovopyranosyl) glycerol (III) and (2S)-1-O-palmitoyl-2-O-(11Z-octadecenoyl)-3-O-(6'-sulfo-alpha D- quinovopyranosyl) glycerol (IV), stearic acid (V), methyl myristate(VI) and palmitic acid (VII). CONCLUSION: Compounds I-VI were isolated from the alga for the first time while I, II and IV are new natural products. I-IV showed activity causing morphological abnormality of P. oryzae mycelia. PMID- 12774371 TI - [Triterpenoid saponins from bark Mitragyna inermis]. AB - OBJECTIVE: To study the constituents of Mitragyna inermis that have an anti-tumor activity on Bel-7402 of hepatic carcinoma. METHOD: The compounds were isolated by column chromatography on silica gel, ODS, Sephadex LH-20 separately and their structures were elucidated by chemical and spectral technology. RESULT: Three quinovic acid glycosides, quinovic acid 3 beta-beta-D-glucopyranosyl-(1-->4) alpha-L-rhamnopyranosyl-28-O-beta- D-glucopyranoside(I), quinovic acid 3 beta-O beta-D-quinovopyranoside(II), quinovic acid 3 beta-O-beta-D-glucopyranoside(III), were obtained. CONCLUSION: I, II were isolated from Mitragyna for the first time. PMID- 12774372 TI - [Studies on chemical constituents from the seed of Trigonella foenum-graecum]. AB - OBJECTIVE: To study the chemical constituents from the seed of Trigonella foenum graecum. METHOD: The compounds were isolated with silica gel chromatography and their structures were identified by physical, chemical properties and spectral analysis. RESULT: Seven compounds were isolated and identified as N,N' dicarbazyl, glycerol monopalmitate, stearic acid, beta-sitosteryl glucopyranoside, ethyl-alpha-D-glucopyranoside, D-3-O-methyl-chiroinsitol and sucrose. CONCLUSION: All the compounds were obtained from this plant for the first time and N,N'-dicarbazyl is a new natural product. PMID- 12774373 TI - [Studies on chemical constituents from the water-soluble part of root of Angelica dahurica]. AB - OBJECTIVE: To study the chemical constituents in the root of Angelica dahurica. METHOD: The compounds were isolated and identified by column chromatography and 1H, 13C NMR data. RESULT: Eleven compounds were isolated and identified. CONCLUSION: Six compounds were isolated from this plant for the first time. PMID- 12774374 TI - [Studies on the chemical constituents in root of Rheum rhizastachyum]. AB - OBJECTIVE: To investigate the chemical constituents from the root and rhizome of Rheum rhizastachyum. METHOD: The compounds were isolated by column chromatography on silica gel, Sephadex LH-20, MCI-gel CHP-20P separately and their structures were elucidated by chemical and spectral technology. RESULT: Six compounds were isolated and identified as chrysophanol, emodin, gallic acid, sucrose, 1-O-beta-D glucopyranosyl-chrysophanols, 1-O-beta-D-glucopyranosyl-emodin. CONCLUSION: All the compounds were obtained from this plant for the first time. PMID- 12774375 TI - [Studies on chemical constituents in herba of Lysimachia davurica]. AB - OBJECTIVE: To study the chemical constituents from the whole plant of Lysimachia davurica. METHOD: The constituents were separated and purified by column chromatograph, and identified by spectral analysis and physical data. RESULT: Six compounds were isolated and identified as triacontanoic acid(I), palmitic acid(II), beta-amyrin(III), stigmasterol(IV), oleanolic acid(V), soya-cerebroside I (VI). CONCLUSION: I, III, VI were isolated from this gene for the first time. PMID- 12774376 TI - [Effects of soy extract on lipid metabolism in ovariectomized rats]. AB - OBJECTIVE: To study effects of soy extract on lipid metabolims in ovariectomized rats. METHOD: 90 Wistar rats were randomly divided into 9 groups: control group, sham group, model group, estrogen group, soy isoflavone group of high dose, soy isoflavone group of low dose, soy extract of high dose, soy extract of low dose, and soy polysaccharde group, 10 rats in each group. Except fer of control and sham groups, the test rats were ovariectomized. One week after operation, the rats were treated with different drugs. Six weeks after operation, the rats were killed, with serum and liver taken, and serumglycerol(sGT), cholesterol(sGC), LDL, HDL and liver homogenate hGT, hGC, measured. RESULT: The level of sGC, LDL in ovariectmized rats increased significantly, compared with that in control and sham groups. In liver both the level of hGT and hGC were higher than that in liver from control and sham groups. Administration of estrogen or soy extract or soy isoflavone could attenuate these in ovariectomized rats, but soy polysacchardes did not have any effects. CONCLUSION: Ovariectomized rats have an imbalance of lipid metabolism, the level of hGT and hGC were increased, and administration of estrogen, soy extracts or soy isoflavone could decrease these changes induced by ovariectomizing. PMID- 12774377 TI - [Influence of the watery extract of jiangzhining decoction on the genetic expression of hepatocyte LDLR of hyperlipidemic rats]. AB - OBJECTIVE: To study the influence of Jiangzhining decoction on the genetic expression of Liver LDLR of the rats suffered from hyperlipemia. METHOD: Laboratory animals were male wister rats with hyperlipemia resulting from high fat feeding. Prescription was the douche of stomach with Jiangzhining decoction (200%) with a dosage of 1.4 g.kg-1, for 15 successive days. Total RNA was extracted from the liver tissue of treated rats and LDLRmRNA was detected by Dot blot hybridization. Expression levels of LDLRmRNA was estimated by a ratio of LDLRmRNA and beta-actin mRNA. RESULT: The difference between expression levels of LDLRmRNA for normal group and those for hyperlipemia group (100% +/- 19% vs 39% +/- 14%) was significant (P < 0.05); and the difference between decoction group (108 +/- 8%) and hyperlipimia group was also highly significant (P < 0.01). CONCLUSION: High fat feeding reduces the expression of liver LDLRmRNA while the decoction can greatly increase it. The study and development of Jiangzhining are significant in preventing and curing cadiocerebral diseases. PMID- 12774378 TI - [Study on sequence difference and SNP pheomenon of rDNA ITS region in F type and H type population of Dendrobium officinale]. AB - OBJECTIVE: To study rDNA ITS sequence differences between F type and that of H type of Dendrobium officinale in main habitat of China. METHOD: The population differences of the rDNA ITS region (including ITS1, ITS2, 5.8S) sequences of D. officinale were studied by the method of DNA sequences analysis. RESULT: There were two different sites between the rDNA ITS sequence of F type and that of H type. One was in ITS1 region, and the other was in 5.8S region. It was proved that there was some relativity between the character of rDNA ITS region and the life type of the populations. The phenomenon of single nucleotide polymorphism (SNP) existed in 5.8S region of rDNA ITS region between F type and H type. The sequences of rDNA ITS region of D. officinale were reported for the first time, and the sequences of ITS region ranged 634 bp (ITS1 231 bp, ITS2 240 bp, 5.8S 163 bp). CONCLUSION: The analysis of rDNA ITS of D. officinale deeply reveal the population differences of D. officinale of F type and H type. PMID- 12774379 TI - [Pharmacognostical identification of "guijiu ([symbol: see text])" in Shosoin of Japan]. AB - OBJECTIVE: To identify the botanical origin of the Guijiu in Shosoin of Japan from Tang Dynasty, and trace its medicinal history. METHOD: Anatomical characteristics of the underground parts of Guijiu in Shosoin were compared with those of Hosta plantaginea and H. ventricosa, and research on the medicinal history of Guijiu was made based on its original identification results and describes in herbals. RESULT AND CONCLUSION: Guijiu in Shosoin of Japan was derived from the underground parts of H. plantaginea and is one of Guijiu used in Tang Dynasty and earlier on. PMID- 12774380 TI - [Pharmacognostic identification of medicinal Asteropyrum plants in China]. AB - OBJECTIVE: To offer evidences for quality control of medicinal Asteropyrum plants. METHOD: Pharmacognostic studies were made through field collection, market investigation, document utilization, comparative morphology and histology. RESULT: The shape and properties, microscopic characteristics in root, rhizome, leaves were worked out. CONCLUSION: Asteropyrum can be distinguished from close relative Coptis plants by microscopic and histology characteristics. Two species in Asteropyrum can also be identified by microscopic and histology characteristics, and the morphological and histology characteristics can be used as evidences for quality control of medicinal Asteropyrum plants. PMID- 12774381 TI - [A novel fingerprint method for quality evaluation of Chinese medicinal plants based on analytical data visualization]. AB - OBJECTIVE: To investigate new method for evaluating the quality of Chinese Medicinal Plants (CMP). METHOD: A visualization technique for representing instrumental analytical data was developed by applying the fundamental of Data Visualization, with Principal Component Analysis (PCA) and spatial projection transformation, original IR spectral data were projected into a low-dimensional subspace so that the dimensionality of original data space was decreased and tiny fingerprint features were extracted. The data set in the subspace was visualized by means of two-dimensional grayscale images. Consequently, the characteristic fingerprint for appraising the quality of CMP was obtained. RESULT: 42 mulberry root-bark samples from three different quality classes were identified with the proposed method, which showed that the fingerprint images had satisfactory resolution and classification accuracy as high as 90.5%. CONCLUSION: The proposed method is a useful technique for appraising the quality of CMP. PMID- 12774382 TI - [Studies on chemical constituents of Cuscuta chinensis]. AB - OBJECTIVE: To study the chemical constituents of the seeds of Cuscuta chinensis. METHOD: The separation was carried out by polyamide and silica gel chromatography, and the compounds were identified by means of physico-chemical and spectroscopic methods. RESULTS: Eight compounds were isolated from the plant and identified as quercetin 3-O-beta-D-galactoside-7-O-beta-D-glucoside (I), quercetin 3-O-beta-D-apiofuranosyl-(1-->2)-beta-D-galactoside (II), hyperoside (III), isorhamnetin (IV), kaempferol (V), quercetin (VI), d-sesamin (VII) and 9(R)-hydroxy-d-sesamin (VIII). CONCLUSION: Compounds IV and VII were isolated from Cuscuta for the first time, and I, II and VIII were characteristic constituents for this vegetable drug. PMID- 12774383 TI - [Studies on chemical constituents from Tibetan medicine wangla(rhizome of Coeloglossum viride var. bracteatum)]. AB - OBJECTIVE: To study the chemical constituents of Tibetan Madicine Wangla. METHOD: Sepearing the chemical constituents by means of chromatography and identifying their structures on the basis of MS and NMR spectra and TLC with authentic samples. RESULT: Eight compounds were isolated as 4-[(4-hydroxyphenyl) methoxy]benzenemethanol(I), 4,4'-dihydroxydiphenyl methane(II), 4,4' dihydroxydibenzyl ether(III), gastrodin(IV), 4-hydroxy benzenemethanol(V), 4 hydroxybenz aldehyde(VI), beta-sitosterol(VII) and beta-daucosterol(VIII). CONCLUSION: All the compounds were obtained from genus Coeloglassum for the first time. The compound I is a new natural product. PMID- 12774384 TI - [Chemical constituents of Hyperricum monogynum]. AB - OBJECTIVE: To investigate the chemical constituents from the aerial parts of Hyperricum monogynum. METHOD: Compounds were isolated by various column chromatography and identified by spectral analysis. RESULT: Ten compounds were isolated and identified as quercetin, quercitrin, hyperoside, rutin, (-) epicatechin, 3,5-dihydroxy-1-methoxy-xanthone, 3,4-O-isopropylidenyl shikimic acid, shikimic acid, daucosterol, and oleanoic acid. CONCLUSION: All compounds were isolated from this plant for the first time. PMID- 12774385 TI - [Studies on chemical constituents of two plants from Costus]. AB - OBJECTIVE: To study the chemical constituents of Costus speciosus and C. tonkinensis (Zingiberaceae) distributed in Yunnan province. METHOD: Chromatography and spectral analyses were used to isolate the constituents and elucidate their structure. RESULT: Six compounds were isolated from the rhizome of C. speciosus and elucidated as diosgenin(1), prosapogenin B of dioscin(2), diosgenone(3), cycloartanol(4), 25-en-cycloartenol(5) and octacosanoic acid(6). Four compounds were isolated from the rhizome of Costus tonkinensis and elucidated as tetracosanoic acid(7), succinic acid(8), beta-sitosterol(9) and daucosterin(10). CONCLUSION: Compounds of 3-6 were obtained from C. speciosus for the first time and compounds of 7-10 were obtained from C. tonkinensis for the first time too. PMID- 12774386 TI - [Studies on the chemical constituents of the stems of Alyxia sinensis (I)]. AB - OBJECTIVE: To demonstrate the chemical constituents of Alyxia sinensis. METHOD: Phytochmical experiment was carried out, using column chromatograph technologies. RESULT: Five compounds have been isolated from the petroleum ether soluble part of the stems of A. sinensis. Their structures have been elucidated respectively as heptatriacontane(1), octatriacontane(2), 20-noatriacontannone(3), 20 nonatriacontanone(4), 20-tetraacontanoe(5), physcion(6), emodin(7), chrysophanol(8), coumarin(9), stigmasterol acetate(10), beta-sitosterol acetate(11), lupeol(12), betulin(13), stigmasterol(14), beta-sitosterol(15), ursolic acid(16), oleanolic acid(17). CONCLUSION: All these compounds are firstly been isolated from A. sinensis. And compound 5-9, 10, 11 are also been separated from Alyxiae genus for the first time. PMID- 12774387 TI - [Structural features of a neutral heteropolysaccharide CPB-4 from Cynanchum paniculatum]. AB - OBJECTIVE: To study the chemical features of CPB-4, a heteropolysaccharide obtained from Cynanchum paniculatum. METHOD: Sugar composition analysis, methylation analysis, partial hydrolysis and carbon-13 nuclear magnetic resonance were used to determine the sugar composition, linkages, main chain, branch chains and branching points. RESULT: CPB-4 is composed of L-arabinose, L-xylose, L rhamnose and D-galactose in closely molar ratios of 0.8:0.2:0.2:1.0. Its main chain is comprised of 1, 5 linked galactose and side chains are comprised of terminal xylose, terminal arabinose, oligosaccharide of arabinose and oligosaccharide of arabinose, rhamnose and galactose. The branching points are located at C-6 and C-2 of galactose. CONCLUSION: CPB-4 is a new heteropolysaccharide from C. paniculatum. PMID- 12774388 TI - [Studies on chemical constituents of Alpinia jianganfeng]. AB - OBJECTIVE: To study the chemical constituents of Alpinia jianganfeng (Zingiberaceae) distributed in Sichuan province. METHOD: Chromatography and spectral analyses were used to isolate the constituents and elucidate their structure. RESULT: Four compounds were isolated from the rhizome of A. jianganfeng and elucidated as kaempferol-3-O-glucuronide(1), docosanoic acid(2), 3-hydroxy-stigmast-5-en-7-one(3) and beta-sitosterol(4). CONCLUSION: All these compounds were obtained from this plant for the first time. PMID- 12774389 TI - [The measurement of tyrosine hydroxylase activity in the brain of conscious rats]. AB - OBJECTIVE: To set the measuring method of tyrosine hydroxylase activity in the brain of conscious rats. METHOD: By using microdialysis and High Performance Liquid Chromatography-Electrochemical Detector system, the 3, 4 dihydrioxphenylalanine (DOPA) formation in the striatum of 6-hydroxdopamine pretreated rats during infusion of an L-aromatic amino-acid decarboxylase inhibitor (NSD1015) was monitored. RESULT: The absence of DOPA in dialysates of 6 hydroxdopamine-pretreated rats, the measurable DOPA and the steady decreasing of 3,4-dihydroxyphenylacetic acid(DOPAC) during infusion of NSD1015 and the disappearance of DOPA after administration of alpha-methyl-rho-tyrosine indicated that the dialyzed DOPA was derived from dopaminergic nerve terminals. After intraperitoneal administration of dopamine receptor agonist apomorphine the DOPA output was deseased. After intraperitoneal administration of dopamine recepter antagonist haloperidol, the DOPA output was increased. The study showed that twenty-four hours ofter implantation of the probe with infusion of 0.01 mmol.L-1 NSD1015, the DOPA level in the striatum of 6-hydroxdopamine-pretreated rats was 0.39 +/- 0.12 pmol/min (X +/- S, n = 5). CONCLUSION: The DOPA concentration in striatal dialysates could be considered as an index of tyrosion hydroxlase activity during infusion of 0.01 mM NSD1015. The method in vivo to monitor tyrosine hydroxlase activity in the brain is reliable. PMID- 12774390 TI - [Effects of soy extract on energy balance in ovariectomized rats]. AB - OBJECTIVE: In order to study on effects of soy extract on energy metabolims in ovariectomized rats. METHOD: 90 Wistar rats were randomly divided into 9 groups: control group, sham group, model group, estrogen group, soy isoflavone group of high dose, soy isoflavone of low dose, soy extract of high dose, soy extract of low dose, 10 rats each group. Beside of control and sham groups, the rest rats were ovariectomized. One week after operation, the rats were treatmented with different drugs, measument of body weigh and feed weigh each week. Six week after operation, the rats were killed, serum were taken, abdomen lipid were removed and weight. RESULT: The ovariectomized rats took more food and got weight gain significantly; Body mess index(BMI), Abdomen lipid weigh and food transform rate in Model group increased significantly than control and sham groups. Administration of estrogen or soy extract or soy isoflavone could block these changes in ovariectomized rats, but soy polysaccharides did not have the effects. CONCLUSION: Ovariectomized rats have imbalance of energy metabolism, weigh gain and accumulation of abdomen lipid; administration of estrogen, soy extracts or soy isoflavone could attenuate these changes induced by ovariectomizing. PMID- 12774391 TI - [Effects of the zhikuofang on the inflammation and cytostatics of the airway model of bronchiectasis]. AB - OBJECTIVE: To explore the effects of Zhikuofang, a TCM prescription, and Ofloxacin on the inflammation and cytostatics of the airway model of bronchiectasis. METHOD: The airway model of bronchiectasis (AMB) was set up and infused with Ps. Aeruginosa. A comparison between the effects of Zhikuofang and Of loxacin on the AMB was made. RESULT: Zhikuofang is better than Ofloxacin in following aspects: lowering the density of inflammation cells in blood, decreasing the volume of tracheal secretion and inhibiting the cytostatics (IL-8 and TNF-alpha) of the trachea tissue, but Ofloxacin is more effective in diminishing the amount of bacteria in trachea flushing liquor. There was no marked difference between them in their histopathy effects on the trachea. CONCLUSION: Zhikuofang probably plays antiphlogistic and bacteriostatic effects by inhibiting the IL-8 and TNF-alpha, resisting secretion, decreasing the inflammation cells and resisting inflammation of trachea. PMID- 12774392 TI - [Modulation of lianbizi injection (andrographolide) on some immune functions]. AB - OBJECTIVE: To study the effects of andrographolide on immune functions and the immune mechanism of its clinical application. METHOD: The amounts of IFN-alpha, IFN-gamma, TNF-alpha, IL-8 in the peripheral blood mononuclear cells(PBMCs) culture supernatants deal with by different concentrations of LianBiZhi (LBZ) injection made of andrographolide were detected by biological activity test or ELISA in vitro. The effects of LBZ injection on macrophage phagocytotic function and natural killer cells cytotoxicity were examined by means of macrophage to phagocytize cock erythrocyte and measurement of lactate dehydrogenase(LDH) activity released from the damaged cells, respectively. RESULT: The LBZ injection could promote IFN-alpha, IFN-gamma, TNF-alpha inductions of PBMCs, but had no effect on IL-8. At the same time, the LBZ injection could not only enhance the phagocytosis activity of peritoneal macrophage from guinea pig to phagocytosis cock erythrocyte, but also augment the cytotoxicity mediated by natural killer cells from PBMCs to damage the K562 cell lines. CONCLUSION: Andrographolide is an immunostimulant agent which can modulate both antigen specific and nonspecific immune function by means of its natural killer cells and macrophage and cytokines induction. PMID- 12774393 TI - [Determination of flavone for Scutellaria baicalensis from different areas by HPLC]. AB - OBJECTIVE: To describe the difference between native and nonative herbs by determining contents of seven kinds of flavone for twenty-five samples from seventeen areas. METHODS: HPLC. Fluid phase: MEOH-H2O-CH3COOH(ICE) (41:59:0.2) and (50:50:0.2). Detection wavelength: 275. RESULTS: The contents of baicalin are 6%-9%, wogenin are 2%-8%, baicalein are 0.1%-1.6%, neobaicalein are 0.01%-0.2%, wogonin are 0.01%-0.3%, visidulin and oroxylin are trace amounts or undetected. CONCLUSION: The native and nonative herbs have no distinct differce in absolute component ratio. The ratio of baicalin and wogenin is under three. The ratio of baicalin and baicalein, baicalin and wogonin is between twenty and fifty. PMID- 12774394 TI - [Study on the detecting methods of the imported materia medica--olibanum]. AB - OBJECTIVE: To analyse the chemical components of the essential oil of Gum olibanum somalilnds and Gum olibanum Ethiopia, and to set up determination methods of their main components. METHOD: Two kinds of essential oil are identified by GC-MS, and assayed by Gas chromatography, using SE-54 as the packing material (column 2.1 m x 3.2 mm), with column temperature starting from 80 degrees C, holding for 1 min, and then rising at the rate of 15 degrees C per minute to 170 degrees C. RESULT: 40 kinds of chemical compounds in the essential oil of Gum olibanum somalilnds and 22 kinds of those of Gum olibanum Ethiopia were identified by GC-MS, the main component in the essential oil of Gum olibanum somalilnds being alpha-pinene, and the main one of Gum olibanum Ethiopia being Octyl acetate 17 batches of samples were determined with the linear range of alpha-pinene being 0-10.80 micrograms, the correlation coefficient being 0.9995, the recovery being 98.16%, RSD being 1.83%; the linear range of Octyl acetate being 0-10.32 micrograms, the correlation coefficient being 0.9996, the recovery being 99.56%, and RSD being 1.36%. CONCLUSION: This study can be used for the setting up of the specification of Olibanum. PMID- 12774395 TI - [Determination of gossypol in cotton root bark by HPLC]. AB - OBJECTIVE: To establish a method for the determination of gossypol in cotton root bark. METHOD: Samples were extracted with acetone, and determined on a C18 column with the mobile phase (Acetonitrile-0.2% phosphoric acid, 85:15) and UV-235 nm detector. RESULT: The recovery rate was 97.6%, RSD 1.59% (n = 5). CONCLUSION: This method can be used for the determination of gossypol in cotton root bark and the content of gossypol in three different species has been determined. PMID- 12774396 TI - [Study on the pathogen and its biological characteristics of opium poppy downy mildew]. AB - OBJECTIVE: To study the pathogen of opium poppy downy mildew and its biological characteristic for further research on the disease. METHOD: Development of the disease was observed systematically in the field. Germination rate of sporangium in different temperature, pH and nutrition was examined with suspending-drop method. Slide-germination method was used to observe its germination in different humidity maintained by different concentration of H2SO4. RESULT AND CONCLUSION: The disease manifests itself in two forms: severely infected plants (systematic infection) and leaf spots (nonsystematic infection). Sporangia of the pathogen are oval or globular, thin walled, smooth, hyaline, with 7.74-16.34 microns diameter in base 1 and 8.34-15.05 microns in base 2.0 ospores are light yellow with 33.87-70.54 microns x 19.34-62.64 microns in base 1 and 36.85-49.68 microns x 42.08-55.76 microns in base 2. Conidiophores are stout, erect, whose branching times and length are different between those in base 1 and those in base 2. Sporangia sprot directly in two hours. Film of water is necessary for sporangium to sprot. The optimum temperature range of sporangium sprot is 12-21 degrees C, the best being 16 degrees C, the pH range is 4.53-9.18 the best optimum at pH 7.38, and the extract of leaf of 1:5 is good for its germination. PMID- 12774397 TI - [Determination of vitamin C in the Vc yinqiao tablet by diffuse reflectance FTIR]. AB - OBJECTIVE: To establish a method to determine Vitamin C in the Vc Yinqiao Tablets. METHOD: Diffuse Reflectance in FTIR was used to determination of Vitamin C in the Vc Yinqiao tablets. RESULT: The content of Vitamin C could be obtained directly, and the relationship between the absorbance and the concen tration was linear. The average recovery rate of Vitamin C was 98.69%, and RSD was 0.33%. CONCLUSION: The method is simple, accurate and reliable, which may well be used for the determination of Vitamin C in the Vc Yinqiao Tablet. PMID- 12774398 TI - [Determination of curculigoside in crude medicine Curculigo orchioides by HPLC]. AB - OBJECTIVE: To establish a method for the determination of curculigoside in Curculigo orchioides, a species of crude medicine. METHOD: The methanol was used as the solvent of exatraction and the curculigoside was extracted from the crude medicine with the method of the ultrasonic vibration. Using Sep-Pak C18 cartridges to purify the solution, the curculigoside was detected by HPLC. Intersil ODS-3(150 mm x 4.6 mm, 5 microns) chromatographic column was used, mobile phase of methanol-water-ice acetic acid (45:80:1) and detect wavelength was set at UV 283 nm. RESULT: The average recovery was 99.2% for the determination of curculigoside, RSD = 1.7% (n = 5). The content range of the curculigoside for 6 kinds of different samples was from 0.11% to 0.35%. CONCLUSION: This method could be used to control the quality of crude medicine C. orchioides. PMID- 12774399 TI - [Determination of trigonelline in Trigonella foenum-graecum by HPLC]. AB - OBJECTIVE: A HPLC method is established to determine the content of trigonelline in Trigonella foenum-graecum. METHOD: The medicinal material was extracted by petholeum ether-ethanol. Asahipak NH2P-50 column was used, mobilephase consisted of acetonitrile-water(75:25) and detection wavelength was set at UV 265 nm. RESULT: The standard curve was linear in the range of 3.68-73.60 micrograms.mL-1 with the correlation coefficient of 0.9999. The average recovery rate and RSD were 97.4% and 1.83% (n = 6) respectively. CONCLUSION: It provides scientific indexes for quality control of T. foenum-graecum. PMID- 12774400 TI - [Chemical constituents from Alyxia sinensis (II)]. AB - OBJECTIVE: To demonstrate the chemical constituents of Alyxia sinensis. METHOD: The constituents were isolated by column chromatography and identified by advanced physical and spectral analysis. RESULT: Eight compounds have been isolated and elucidated as bauereny acetate(18), scopletin(19), liriodendrin(20), pinoresinol-di-O-beta-D-glucopyranoside(21), daucosterol(22), flaxetin(23), esculin(24), aseculin(25). CONCLUSION: These compounds were found from the plant for the first time, and compound 20,21,23-25 were found from Alyxiae genis for the first time, and compound 18 is firstly been isolated from natural source. PMID- 12774401 TI - [Studies on chemical constituents in buds of Artemisia scoparia]. AB - OBJECTIVE: To investigate the chemical constituents in flower buds of Artemisia scoparia. METHOD: The constituents were separated and purified by means of various chromatographic techniques, and their structures were determined on the basis of physical and chemical properties and spectral data. RESULT: Four flavones were isolated and their structures were identified as cirsilineol (I), cirsimaritin(II), arcapillin(III) and cirsiliol(IV) respectively. CONCLUSION: Compounds I, III and IV were isolated from this plant for the first time. PMID- 12774402 TI - [Studies on second metabolites of an endophytic fungus (II)]. AB - OBJECTIVE: To study the chemical constituents from the cultured mycelia of a fungus Cephalosporium which accelerate the growth of plant. METHOD: The constituents were isolated by column chromatography and identified by advanced physical and spectral analysis. RESULT: Eleven compounds including 3-isopropyl-6 (1-methylpropyl) piperazine-2,5-dione(I), choline sulfate(II), 2-[(2-hydroxy tetracosanoyl) amino]-1,3,4-octadecatriol(III) were isolated and identified. CONCLUSION: Compound I, II were isolated from Cephalosporium genus for the first time. PMID- 12774403 TI - [Studies on chemical constituents of Galeopsis bifida]. AB - OBJECTIVE: To investigate the chemical constituents of Galeopsis bifida. METHOD: Various chromatographic techniques were employed for isolation and purification of the constituents. The structures were elucidated by chemical and spectral analyses. RESULT AND CONCLUSION: Ten compounds were isolated and identified as chrysoeriol(1), 5,7-dihydroxy-3',4'-dimethoxyflavone(2), hederagenin(3), daucosterol(4), martynoside(5), ajugoside(6), 8-acetyl harpagid(7), 5,7,4' trihydroxy-3'-methoxyflavone-7-O-beta-D-glucoside(8), 5,6,7,4' tetrahydroxyflavone-7-O-beta-D-glucoside(9) and 5,7,3',4'-tetrahydroxyflavone-7-O beta-D-glucoside (10). Compound 6, 7 were found from genus Galeopsis for the first time and rest of compounds were found from G. bifida for the first time. PMID- 12774404 TI - [Studies on chemical constituents of root of Cichorium intybus]. AB - OBJECTIVE: To study the chemical constituents in the root of Cichorium intybus. METHOD: The compounds were isolated and identified by column chromatography and NMR, IR, MS data. RESULT: Twelve compounds were isolated and identified. CONCLUSION: 2,3,4,9-tetrahydro-1H-pyrido-(3,4-b)indole-3-carboxylic acid was isolated from the Cichorium genus for the first time, azelaic acid and daucosterol were isolated from the the plant for the first time. PMID- 12774405 TI - [In vitro study on realgar induced apoptosis of all-trans acid resistant acute promeylocytic leukemia cell line (MR2)]. AB - OBJECTIVE: To acquire a deep understanding of the possible mechanisms of realgar in the treatment of acute promyelocytic leukemia (APL). METHOD: All-trans retinoic acid (ATRA) resistant APL cell line MR2 was used as in vitro model. The effect of realgar on MR2 cell was observed by watching cell viability, cell growth, and by using Methy thiazolyl tetrazolium (MTT) assay, cell morphology, DNA gel electrophoresis and flow cytometry assay. RESULT: The viability and growth of MR2 cell were inhibited after the treatment, to some extent, in a dose and time dependent manner. After being treated with realgar, MR2 cell presented morphologically some features of apoptotic cells such as intact cell membrane, chromatin condensation and nuclear fragmentation, and apoptotic body could be found by electron microscopy as well. Sub-G1 cells were observed by flow cytometry, as well as Annexin V FITC+/PI-cells. DNA ladder could be found by DNA gel electrophoresis. CONCLUSION: Realgar can induce apoptosis of ATRA resistant APL cell line MR2, Which shows the therapeutic effect of realgar on APL may be different from that of ATRA. PMID- 12774406 TI - [Study on antiinflammatory effect of a compound TCM agent containing ant extractive in animal models]. AB - OBJECTIVE: To study the antiinflammatory effect of a compound TCM (Traditional Chinese Medicine) agent on animal models. The agent contains ant extractive and a blent of three herbal products, herba epimedii, fructus cnidii, and fructus lycii. METHOD: Three animal models to induce experimental inflammation in rats, including carrageenin--induced paw edema, cotton-ball granuloma and adjuvant induced arthritis, were chosen to study the antiinflammatory effect of the TCM agent. RESULT: The TCM agent showed a marked inhibitory effect on edema induced by all three types of inflammation in rats, the inhibitory rate of the TCM agent at the dose of 0.20, 0.40 and 0.80 g.kg-1 in granuloma model bing over 25% at 1 hour post oral administration, and being 23.8%, 22.7%, 39.7% at 6 hour. In addition, the TCM agent also showed a significant preventive as well as therapeutic effect on adjuvant induced arthritis in rats, and improved the pathological changes of the animal joints with the induced arthritis. CONCLUSION: TCM agent has significant antiinflammatory effects on the three above mentioned animal models. PMID- 12774407 TI - [Effects of epimedium total flavonoids phytosomes on preventing and treating bone loss of ovariectomized rats]. AB - OBJECTIVE: To observe the effects of Epimedium total Flavonoids Phytosomes on preventing and treating bone-loss of the castrate osteoporosis rat model. METHOD: The osteoporosis model was established with 4-month-odl panther's rats, their ovaries on both sides castrated. Dual energy X-ray scanning was used to determine the bone density, and immunity and ELASA were used to assay concentration of estradiol and IL-6 in serum respectively, then determine their effect. RESULT: The BMP and E2 of high dosage group nilestriol group and normal group are higher than those of model group (P < 0.01), while their content of IL-6 is apparently lower than that of model group(P < 0.01). CONCLUSION: The osteoporosis model was established successfully and the using of EFP can improve the bone density, enhance E2 level and decrease the IL-6 concentration in serum. PMID- 12774408 TI - [Aspects of the Moscow fluorographic services status and possible approaches to its reorganization]. AB - Due to the fact that modern digital fluorographic plants have been introduced into medical practice, it is necessary to elaborate new organizational and technological forms of work during fluorographic survey. For this, 1-2 Moscow okrugs completely equipped with digital fluorographic plants should be used to examine their actual capacities and on this basis to develop guidelines for detection of chest diseases by applying this new technological equipment. While organizing mass surveys by using digital fluorographs, it is necessary to take into account the definite conditions: health care facilities must be equipped with a uniform digital fluorograph; for continuity in work and information exchange, the software of all fluorographs must be unified within the city; a personified electronic fluographic card catalogue must be provided by identifying decreed contingents and tuberculosis risk groups; digital fluorographs must be fitted with the programmes that permit one to keep records and accounts in accordance with the approved official statistical forms; the database must be compatible in all users for prompt exchange of information on modem communication and for transmission of the results of a survey on diskette carriers; a fluorographic center wherein the fluorograms of individuals with suspected tuberculosis should be collected and deciphered (by modem communication or on diskette information carriers) must be set up on the basis of one of the tuberculosis dispensaries. PMID- 12774409 TI - [Ways of enhancing the efficiency of tuberculosis-controlling measures and reducing their cost]. AB - As of now, there are prerequisites for enhancing the efficiency of tuberculosis controlling measures and for reducing their cost by utilizing material and personnel resources in priority areas. These include: to detect patients with tuberculosis by using current computer technologies, to apply digital X-ray plants, to recruit general practitioners for preventive measures against tuberculosis, to reduce the cost of treatment in patients with tuberculosis by decreasing hospital stay. Introduction of currently available procedures substantially lowers the cost of antituberculous aid and enhances its efficiency. PMID- 12774410 TI - [Criteria for the clinical cure of tuberculosis of the eye]. AB - The results of examination were analyzed in 314 patients with tuberculosis of the eye, including 120 patients having its inactive phase. Tuberculin skin tests were performed in 31 patients; fluorescence angiography and photostress tests were conducted in 122 patients, and 120 patients underwent lacrimal fluid crystallography. To define the cure rate in tuberculosis of the eye, it is necessary to have a complex of criteria: no clinical signs of active tuberculous inflammation in the ocular tunics and environments; no clinical, X-ray, and laboratory manifestations of tuberculosis of other organs; no focal and significant overall reaction to tuberculin injected in doses of 2 and 50 TU. If there are contraindications to tuberculin skin tests to conclude whether the disease is cured, it is necessary to use fluorescence angiography of the fundus of the eye, lacrimal fluid crystallography, and/or photostress tests in combination with biochemical and immunological blood studies. PMID- 12774411 TI - [Extrapulmonary tuberculosis in West Siberia]. AB - Nine hundred and seventy-three cases of extrapulmonary tuberculosis were analyzed by using the statistical reports from 14 West Siberian areas supervised by the Novosibirsk Research Institute of Tuberculosis, Ministry of Health of the Russian Federation. In 1999, extrapulmonary tuberculosis mortality again increased in Siberia, amounting to 4.4 cases per 100,000. Its incidence was 29.95 per 100,000. In the mortality structure, urogenital tuberculosis held the lead, peripheral lymph nodal tuberculosis and osteoarticular tuberculosis occupied the second and third places, respectively. Cases of advanced and complicated tuberculosis continue to be encountered; one of them is given as an illustration. PMID- 12774412 TI - [Informative value of the major component technique in the assessment of the epidemic tuberculosis situation]. AB - The paper describes a method for comprehensive analysis of the epidemic situation which is based on the major component technique. The algorithm proposed for comprehensive assessment is highly effective in the retrospective analysis and in the analysis of the current material which includes a great deal of various characteristics that are at first sight occasionally unrelated. The use of the algorithm is shown by the example of analyzing 19 statistical variables that characterize the epidemic situation and different aspects of the work of a tuberculosis-controlling facility in 30 districts of the Novosibirsk Region in a period of 15 years (1985-1999). During processing, the variables fall into two groups: a group of variables that characterize the quality of antituberculosis work and a group of those reflecting the epidemic situation, which were used to divide the districts of the region into several groups. PMID- 12774413 TI - [Enzyme immunoassay and immunochromatographic assay in the differential diagnosis of tuberculosis and cancer of the respiratory organs]. AB - The informative value of immunodiagnostic assays aimed at identifying specific antibodies in the peripheral blood was studied. Indirect solid-phase enzyme immunoassay (Antitub test system, Moscow) and rapid immunochromatographic assay (TB-CHECK, France) were used. Fifty-six diagnostic patients and 18 healthy individuals were examined. The diagnosis of active tuberculosis was verified in 29 patients (Group 1) and that of lung cancer was in 27 patients (Group 2). The detection rate of and the level of antituberculosis antibodies in patients with active tuberculosis greatly differed from those in patients with lung cancer (p < 0.01). In the diagnostic patients, the sensitivity of enzyme immunoassay was 79.3%, its specificity was 88.9; the sensitivity and specificity of the rapid assay were 72.4 and 96.3%, respectively. The high specificity of the immunochromatographic assay in patients with lung cancer may reduce the number of false-positive results of the enzyme immunoassay. The concurrent use of both methods is an essential adjunct to the clinical examination of diagnostic patients, it makes it possible to reduce the time of pre-examination of cancer patients and to restrict a group of patients who need for invasive studies to specify a diagnosis. PMID- 12774414 TI - [Role of test therapy with broad-spectrum antibiotics in the diagnosis of nonbacterial tuberculosis]. AB - Test therapy with broad-spectrum antibiotics in the diagnosis of abacillary tuberculosis is indicated in the acute onset of the disease, in focal and infiltrative lung tissue changes, marked pulmonary symptoms (cough, auscultative lung changes, fever). Test therapy can be performed at any stage of diagnosis: in the outpatient setting, in a somatic or tuberculosis hospital. During test therapy, it is irrational to frequently change antibiotics or to use drugs having antituberculous activity (such as fluoroquinolones, rifampicin, most aminoglycosides). Positive X-ray changes after a course of test therapy with non specific antibiotics is a diagnostic criterion which suggests that the disease of non-tuberculosis etiology. PMID- 12774415 TI - [Specific features of the course of pulmonary tuberculosis concurrent with liver pathology]. AB - The study was undertaken to reveal clinical, biochemical, morphological, and immunological features of pulmonary tuberculosis concurrent with liver pathology. Patients with pulmonary tuberculosis who suffered from liver diseases were found to have more marked clinical signs, such as subfebrile temperature, weight loss, elevated levels of aminotransferases and gamma-glutamyltransferase. Morphological studies of liver biopsy specimens indicated that the rates of necrosis and inflammatory infiltration were higher in patients with chronic hepatitis C (CHC) and chronic hepatitis B (CHB) and CHC and substantially lower in CHB and alcoholism. Impaired immunity involved primarily the phagocytic link. PMID- 12774416 TI - [Some specific features of life quality in patients with pulmonary tuberculosis]. AB - The paper presents the results of examination of two groups of male patients with pulmonary tuberculosis: 152 males with fibrosocavernous pulmonary tuberculosis and 123 males with infiltrative pulmonary tuberculosis; which included physical examination, X-ray, laboratory, and functional studies, multifactorial psychological personality testing, survey using a special questionnaire. The functional parameters of health have been found to be worse in male patients with chronic pulmonary tuberculosis. Analyzing the social situation revealed no statistically significant differences in males with chronic or first diagnosed pulmonary tuberculosis. The psychological characteristics were more impaired in male patients with chronic pulmonary tuberculosis. By recognizing that life quality is an integrative indicator of the functional parameters of health and the social and psychological parameters of living standards and life way, life quality in all male patients with pulmonary tuberculosis may be considered to be low. Life quality in male patients with chronic pulmonary tuberculosis is still worse than in those with first diagnosed pulmonary tuberculosis. PMID- 12774417 TI - [The body's adaptation and responsiveness and their impact on the outcome of exudative pleuritis]. AB - The authors studied homeostasis in 182 patients, including 126 patients with exudative pleuritis of tuberculous etiology and 56 with parapneumonic pleuritis. They determined the adaptive reactions of the body by the leukograms and the types of its responsiveness, established their relationship with other homeostatic parameters. Impaired responsiveness in patients with exudative pleuritis substantially affected the efficiency of etiotropic treatment. At the end of the basic course of therapy, the rate of marked residual pleural changes was 84.6% in patients with tuberculous pleuritis (Subgroup 1). Activation pathogenetic therapy with methyluracil promoted a reduction in the number of patients with significant residual pleural changes (32.8% in Subgroup 2). PMID- 12774418 TI - [Causes of ineffective treatment of tuberculosis]. PMID- 12774419 TI - [Enhancing the efficiency of treatment in patients with multidrug resistant pulmonary tuberculosis by a combination of surgery and adequate chemotherapy]. AB - The outcomes of surgical treatment were analyzed in 68 patients with multidrug resistant pulmonary tuberculosis. Of them, 38 patients (Group 1) received preoperative and postoperative chemotherapy with antibacterial drugs by the WHO standard treatment regimens, 30 patients (Group 2) used reserve antibacterial drugs. Chemotherapy and surgical interventions yielded the following results: 84.3 and 93.3% of clinical efficiencies in Group 1 and 2, respectively. The use of reserve antibiotics enhances the efficiency of surgical treatment in patients with multidrug-resistant pulmonary tuberculosis. PMID- 12774420 TI - [Prediction of the effectiveness of preoperative preparation of patients with fibro-cavernous pulmonary tuberculosis]. AB - Sixty-three patients with fibrocavernous pulmonary tuberculosis were examined before and 1 month after preoperative therapy with antituberculosis agents. The time course of changes in the clinical and X-ray characteristics of the severity of the disease were compared with the alterations of different biochemical indices, including acute-phase proteins, proteolysis inhibitors, intoxication severity. It was demonstrated that improvement of clinical and X-ray characteristics of the severity of a process after preoperative preparation could be predicted with an accuracy of 83% from the baseline serum concentrations of albumin and ceruloplasmin. The accuracy of prediction does not increase when the levels of protease inhibitors, C-reactive protein, adenosine deaminase, medium weight molecules are simultaneously studied. The likelihood of the clinical and X ray characteristics becoming better is higher with enhanced ceruloplasmin activity and decreased albumin levels. PMID- 12774421 TI - [Pulmonary tuberculosis in former prisoners]. AB - The examination of 123 former prisoners with pulmonary tuberculosis treated in the Republican Tuberculous Hospital in 1998-2000 versus 37 control patients demonstrated that the specific pulmonary process is revealed at younger age (20 39 years). The treatment effect in former prisoners by percentage of cavern closure is 1.8 times worse than in the controls. This can be explained by significantly higher occurrence of cases with drug-resistant M. tuberculosis and associated diseases. PMID- 12774422 TI - [Tuberculosis and maternity]. PMID- 12774423 TI - [Direct genetic analysis of the rifampicin resistance of M. tuberculosis isolates in sputum samples]. AB - The paper shows a rapid method for diagnosing the resistance of Mycobacterium tuberculosis to rifampicin in the testing of clinical sputum samples. The sputum samples from 12 patients ineffectively treated for pulmonary tuberculosis were treated by the immunomagnetic mycobacterial separation technique; polymerase chain reaction was used to perform the amplification and direct sequencing of the gene fragment rho poB by identifying the mutations responsible for mycobacterial rifampicin resistance. Other equal parts of the same sputum samples were cultured on liquid medium for 5 days and subsequently examined in the same manner and also cultured on the Lowenstein-Jensen solid medium, followed by the determination of rifampicin sensitivity by the routine procedure. Routine examination revealed 7 cases of rifampicin resistance. Short-term (5-day) cultivation of sputum samples, followed by a molecular genetic study, also established rifampicin resistance in all the 7 cases of the 12 tested samples. PMID- 12774424 TI - [The new WHO reference guides (modules) on tuberculosis control at the community level]. PMID- 12774425 TI - [Tuberculosis in the elderly at present]. PMID- 12774426 TI - [Cerebral tuberculoma in a patient with pulmonary tuberculosis]. PMID- 12774427 TI - [The 80th anniversary of the phthisiopulmonology Department of the I. M. Sechenov Moscow Medical Academy]. PMID- 12774428 TI - [Therapy of patients with end-stage pulmonary disease. Review of lung transplantation based on four cases of our own]. AB - The pathogenesis of infiltrative pulmonary diseases is largely unknown, and their final outcome is variable. These inflammatory processes may involve the alveoli and the pulmonary interstices, and they may lead to pulmonary fibrosis. The development of end-stage pulmonary disease is irreversible. Patients with respiratory failure have a poor prognosis. For young patients at the end-stage of their pulmonary disease, lung transplantation is a promising therapeutic modality. The therapeutic protocols for unilateral or bilateral lung transplantation and heart-lung transplantation were established only in the last 10 years. In Hungary there is no center yet performing lung transplantation. This therapy is available only for those who could be effectively rehabilitated with this method. Four patients 27-35 years of age suffering pulmonary fibrosis, despite their young age developed honey-comb lungs with therapy resistant respiratory failure. No other organic symptoms were detected by detailed examination, so their predicted survival was dependent on their pulmonary status alone. Cases of four young patients, who were effectively treated with lung transplantation, are discussed from pre-operative preparations to post-operative follow-up, with the review of the literature. PMID- 12774429 TI - [Tibolone therapy in postmenopausal women with a history of many risk factors]. AB - INTRODUCTION: The changes of the indications and the use of hormone replacement therapy have entered into the focus of interest. AIMS: The author studied whether hormone replacement therapy can be used in postmenopausal patients with high risk factors in their history (thromboembolic disorders, diabetes mellitus, hypertension, stroke, cardiovascular disease, pulmonary embolism). METHODS: Tibolone 2.5 mg daily was prescribed. Data of 124 patients with the above conditions were analized at the beginning and at 12, 24, 36 months follow-up visits. The frequency of exacerbation of the anamnestic risk factors, menopausal complaints, laboratory tests and instrumental tests (transvaginal sonography, mammography, bone densitometry), as well as efficacy and side effects of the therapy were studied. RESULTS: During the 36-months follow up tibolone therapy effectively reduced menopausal complaints, caused no increase in the anamnestic risk factors, or serious side effects and did not worsened the results of the laboratory tests. CONCLUSION: In the author's opinion hormone replacement therapy with tibolone can be safely performed in patients with high risk factors in the history. PMID- 12774430 TI - [Prevalence of the alpha-1 antitrypsin phenotype in Hungary]. AB - INTRODUCTION: Alpha-1-antitrypsin deficiency is one of the most common inherited metabolic disorders. The presence of PiZ and PiS alleles alpha-1-antitrypsin deficeing are associated with. The geographical distribution of PiZ and PiS is heterogenous in Europe. AIM: The aim of the study was to investigate the frequency of alpha-1-antitrypsin alleles with isoelectric focusing in 789 healthy Hungarian citizens and to compare the results to those obtained in other European countries. The measurements were done in the Laboratory of the Institute of Experts in Forensic Medicine. RESULTS: The frequency of PiM, PiZ and PiS alleles were 97.2%, 0.95% and 1.65% respectively. Therefore, the hypothetical prevalences of PiZZ and PiSS alleles are 1:10526 and 1:6060, respectively. CONCLUSION: These results do not differ significantly from those found in the neighbouring countries. Further investigation is needed to elucidate the possible role of alpha-1-antitrypsin phenotypes in the pathogenesis of chronic liver disorders and lung emphysema. PMID- 12774431 TI - [Assessment of cerebrovascular reactivity using the breath-holding test in hypertensive and normotensive adolescents--preliminary results]. AB - AIM: Authors report on preliminary results of cerebral vasoreactivity-tests performed in frame of the Debrecen Hypertension Study among hypertensive and healthy adolescents. METHODS: Investigators measured blood flow velocities before and after 30 seconds of breath holding using transcranial Doppler sonography. RESULTS: Blood flow velocities at rest were significantly higher in the hypertensive that in the healthy adolescent group. Breath holding induced less intensive cerebral vasoreactivity response in the hypertensive than in the normotensive group. CONCLUSION: It is concluded that altered arteriolar function of the cerebral arterioles can be detected already in this early stage of hypertension. PMID- 12774432 TI - [Health economics analysis of cervical cancer screening]. AB - INTRODUCTION: To reduce the high mortality rate of cervical cancer there are organized, nation-wide mass-screening programmes. AIM: To assess the screening rate, the cost of screening and treatment and to calculate the expected epidemiological and economic gain and cost-effectiveness of mass-screening programme. METHODS: The data derive from the financial database of the National Health Insurance Fund of Hungary from 2001. To assess the screening rate the authors used the code "No. 29601 cytological examination for screening" of out patient care. The cost of treatment includes the cost of out-patient care, the acute and chronic inpatient care, the subsidies of medicines' prices and the expenditure on disability to work (including sickness-pay). The expected benefits of the screening programme were modelled with changing the screening interval. RESULTS: The screening rates for 1999, 2000 and 2001 were 14.5%, 16.2% and 15.6% respectively, while the 3 year screening rate for 1999-2001 were 35.7%. The cost of treatment of cervical cancer were around 1 billion Hungarian forint in 2001. The cost of one life saved according to the current screening strategy was 16.6 million Hungarian forints (57.792 USD) with a successful screening programme, while with a less successful program it was 33.8 million Hungarian forint (118.093 USD). The cost of one life year gained according to the current screening strategy was 0.7 million Hungarian forints (2.513 USD) with a successful screening programme, while with a less successful program it was 1.5 million Hungarian forint (5.134 USD). CONCLUSION: It is important to increase the screening rate. With increasing the screening interval for women aged between 25 65 from 1 year to 2 or 3 years, it improves the cost-effectiveness of screening programme. PMID- 12774433 TI - Detecting recent human immunodeficiency virus type 1 infection: why and how? PMID- 12774434 TI - Free speech in the workplace. PMID- 12774435 TI - BD Clinical Laboratory Solutions' president, William A. Kozy, actuates "black belt" quality and "next-gen" planning. Interview by Celia Stevens. PMID- 12774436 TI - Rapid diagnostic testing in microbiology. PMID- 12774437 TI - Integrated automated task targeted analyzer systems. An antidote to front-end processing. PMID- 12774438 TI - HealthSouth woes may spell opportunities for providers. PMID- 12774439 TI - IT-focused, consumer-driven health care offers potential for big savings. PMID- 12774440 TI - Service line strategy provides hospitals with a competitive edge. PMID- 12774441 TI - New strategies required to improve hospital-physician alignment. PMID- 12774442 TI - E-newsletters showcase new resources and build continuing relationships with consumers. PMID- 12774443 TI - Detailed report on physician and patient use of the Web. PMID- 12774444 TI - The physician relationship: key to patients' reliance on your site. PMID- 12774445 TI - Integrating Web services and telephone call center boosts effectiveness. PMID- 12774446 TI - Online billing systems beginning to emerge. PMID- 12774447 TI - New report pegs Americans' use of Web for health information at only 16%. PMID- 12774448 TI - Absence of myocardial release of troponin T after coronary bypass surgery on a beating heart. AB - BACKGROUND: Coronary artery bypass grafting (CABG) operations in connection with cardiopulmonary bypass (CPB) appear to be associated with a number of side effects including trauma, cognitive dysfunction and myocardial damage. Accordingly, a current interest in performing CABG on a beating heart begins to emerge. This study examines the premise that conducting CABG on a beating heart limits the extent of myocardial injury and other complications. METHODS: Forty five consecutive patients underwent CABG on a beating heart (group A, 12 patients) or in connection with CPB (group B, 33 patients). Inclusion criteria were poor left ventricular function and evolving myocardial ischemia or infarction. Results were assessed primarily on the basis of clinical outcome. In addition, measurement of plasma levels of troponin T (TnT), creatine kinase MB (CK-MB) and lactate dehydrogenase (LD) was conducted in 12 patients of group A preoperatively and 24 h after completion of surgery. These biological data were compared with those from control patients who underwent CABG in connection with CPB within the same time span. RESULTS: All patients in groups A and B survived the CABG procedure and those on a beating heart maintained an excellent perioperative hemodynamic measurements. The mean bypass time was 75 +/- 21 min and the mean cardiac standstill was 40 +/- 17 min. The intensive care unit stay was for group A: 18 +/- 4 h, group B: 48 +/- 12 h; and the total hospital stay was for group A: 6 +/- 1 days, group B: 8 +/- 3 days. Angiographic studies showed good anastomatic patency in both groups. Postoperative low output syndrome as indicated by the need of ionotropic drugs for more than 24 h was demonstrated in 4% and 6% of groups A and B, respectively. Limitation of myocardial injury in group A was demonstrated by the minimal increase in postoperative TnT levels (16.0 +/- 0.9 versus 30 +/- 8.0 pg/ml). A similar pattern of changes was observed with other infarction markers including CK-MB and LD. Contrastingly, the pre- and post-operative values of TnT in group B were 18 +/- 1.6 and 790 +/- 140 pg/ml, respectively. CONCLUSIONS: CABG on a beating heart shares many of the positive features of CPB with a distinct advantage of eliminating the intraoperative myocardial ischemia. PMID- 12774449 TI - Influence of C1-esterase inhibitor on tissue oxygenation of jejunal mucosa during endotoxemia. AB - BACKGROUND: The complement system has been shown to play an important role in the pathogenesis of microcirculatory disturbances in trauma and sepsis. The intestinal mucosa is the most susceptible portion of the gut to impaired perfusion and oxygen delivery. The objective of this study was to investigate the effects of C1-esterase inhibitor (C1-INH) on arterial oxygenation (PaO2) and tissue oxygenation (PtiO2) of jejunal mucosa during endotoxemia. METHODS: Eighteen anesthetized and ventilated rats were laparotomized and a jejunal portion was exteriorized and fixed on a plexiglass stage. The jejunum was punctured and a Clark type microcatheter PO2 probe and a micro thermocouple were placed on the mucosa in order to measure PtiO2. The animals were randomly assigned to receive one of the three treatments: infusion of Escherichia coli lipopolysaccharides (LPS) without C1-INH pretreatment (LPS group); or infusion of LPS with C1-INH pretreatment (C1-INH group); the control group (n = 6) without treatment of either C1-INH or LPS. The mean arterial pressure (MAP), heart rate (HR), PaO2 and PtiO2 were measured at baseline, 60 and 120 min after induction of endotoxemia. RESULTS: Hemodynamic parameters (MAP, HR) in all the three groups showed no significant changes during the study period. PaO2 significantly decreased in the LPS group. This decrease could be attenuated by pretreatment with C1-INH. The mucosal PtiO2 of the jejunum in the control group remained stable. It significantly decreased in the LPS and in the C1-INH groups without showing a significant difference after 120 min of endotoxemia. CONCLUSIONS: Pretreatment with C1-INH was able to diminish a decrease in PaO2 during endotoxemia, indicating that pulmonary injury was attenuated. Endotoxin-induced tissue hypoxia of the intestinal mucosa could not be prevented suggesting a minor involvement of complement activation in this pathophysiological process. PMID- 12774451 TI - An inexpensive alternative approach to bone compression analyses. AB - A new method for testing the compression strength of vertebrae was developed and implemented using minimal resources and time expenditure. The device digitally measured and analyzed the torque and force necessary to load rat lumbar and thoracic vertebrae to failure. The system was calibrated using a series of known lead masses. Similar to other standardized force/compression and torque devices, this method has been shown to provide reliable and reproducible results. Furthermore, although vertebrae were the only specimens used to collect data in these initial studies, this measurement system could be altered quite easily to allow for a variety of test materials. PMID- 12774450 TI - Thrombin contraction of vascular smooth muscle: implications for vasospasm. AB - Thrombin contributes to hemostasis by activating platelets, the formation of fibrin, and contraction of the injured vessel. These effects are mediated through the proteolytic activity of thrombin. We hypothesized that thrombin may have a role in vasospasm after arterial injury and examined the physiologic and cellular signaling events of thrombin in intact vascular smooth muscles. Thrombin stimulation of strips of bovine carotid artery smooth muscle led to contractions which relaxed with the addition of the nitric oxide donor, sodium nitroprusside. However, washout of the thrombin and SNP resulted in the re-generation of force. This was not observed with other agonists such as endothelin, thromboxane analogues, or serotonin. Using two-dimensional immunoblotting we demonstrate that thrombin stimulation leads to increases in the tyrosine phosphorylation of 4 proteins, three different isoforms of P44 mitogen activated protein kinase (MAPK) and one isoform of P38 stress activated protein kinase (SAPK). Activation of P38 SAPK leads to activation of MAPKAP kinase-2 and a major substrate protein of MAPKAP kinase-2 is the small heat shock protein, HSP27. HSP27 has been implicated in mediating smooth muscle contraction. These data suggest that in the setting of arterial injury, thrombin-induced contraction may supercede over short acting vasorelaxants such as NO resulting in vasospasm. In addition to stress, physiologic substances such as thrombin, activate SAPKs leading to increases in the phosphorylation of HSP27. Thus, thrombin may play a central role in hemostasis after vascular injury and in the pathologic responses to plaque rupture and thrombosis in atherosclerosis. PMID- 12774452 TI - Acute appendicitis in the elderly. An analysis of 47 patients over 80 years of age. AB - OBJECTIVE: To correlate the preoperative findings, perforation rate and the mortality rate after appendicectomy in the elderly age. DESIGN: Retrospective study. SETTING: Teaching hospital, Sweden. SUBJECTS: Forty-seven patients over 80 years of age who have undergone acute appendicectomy. MAIN OUTCOME MEASURES: Operative and postoperative outcome after acute appendicectomy in the elderly. RESULTS: The complication rate after appendicectomy was statistically significantly increased to 28% as well as the mortality increased to 8.5% in the elderly patients when compared to the other age group. The perforation rate was also statistically significantly increased to 64% when compared to the same group. CONCLUSIONS: Our study shows that at the time of clinical symptoms, inflammatory parameters in acute appendicitis in the elderly do not differ from the young patient, but patients older than 80 years of age have a high risk of perforation, morbidity and mortality. PMID- 12774453 TI - Helicobacter pylori products upregulate neutrophil superoxide anion production. AB - Helicobacter pylori-neutrophil interactions may play a pathogenic role in H. pylori-induced gastritis and peptic ulcer disease. To understand these interactions, we explored the effects of H. pylori-derived products on neutrophil chemotaxis and superoxide anion production. H. pylori bacteria were cultured and supernatants fractionated. Neutrophil chemotactic activity was confirmed in the crude supernatants and in one fractionated peak corresponding to a previously described neutrophil chemotactic factor. H. pylori-derived crude supernatant, sonicate and all chromatography-derived peaks failed to directly stimulate neutrophil superoxide anion production. However, after pretreatment with sonicate, neutrophils demonstrated increased superoxide anion production (priming) following subsequent exposure to the secretagogue fmet-leu-phe. These results suggest that H. pylori products may attract neutrophils to the gastric mucosa without initially stimulating superoxide anion production or tissue injury. Oxygen radical-mediated gastric mucosal injury may subsequently result when these primed neutrophils undergo additional stimulation by as yet unidentified factors. PMID- 12774454 TI - The effect of keratinocyte growth factor-2 on esophagogastric anastomotic wound healing in rats. AB - BACKGROUND AND OBJECTIVES: Esophagogastric anastomotic leaks complicate 5-20% of esophagectomies for esophageal cancer, and they are responsible for approximately one-third of perioperative deaths. Growth factors enhance healing in several wound healing models. Keratinocyte growth factor-2 (KGF-2) is a newly identified growth factor that accelerates skin incisional healing. An animal experiment was done to test the hypothesis that KGF-2 would enhance esophagogastric anastomotic wound healing. METHODS: Forty rats had single layer esophagogastric anastomoses constructed using interrupted 7-0 polypropylene sutures. In the experimental group (20 rats) 1 mg of KGF-2 was administered by intraperitoneal injection on the day of surgery, and for three consecutive days thereafter. Placebo (buffer solution only) was given to the control rats (20 rats). Rats were sacrificed on the fourth postoperative day and their anastomoses were excised, mounted in a tensiometer, and distracted at 10 mm/min to measure breaking strength. After that, anastomotic tissue was subjected to hydroxyproline analysis (an indicator of wound collagen). Skin and fascial wounds were also tested in a tensiometer. RESULTS: There were two anastomotic leaks in the KGF-2 treated rats and none in the control rats (p = 0.26, not significant). Esophagogastric anastomotic breaking strength was 1.00 +/- 0.61 N in the KGF-2 treated rats and 1.58 +/- 0.62 N in the control rats (p = 0.017). Esophagogastric anastomotic tissue hydroxyproline concentration was 95.9 +/- 19.9 nmol/mg in the KGF-2 treated rats and 114.1 +/- 38.0 nmol/mg in the control rats (p = 0.10, not significant). Skin and fascia breaking strengths were also lower in the KGF-2 rats, but the difference was not significantly different (KGF-2 skin -0.97 +/- 0.48 N, Control skin -1.46 +/- 0.89 N, p = 0.064; KGF-2 fascia -3.60 +/- 2.06 N, Control fascia 4.67 +/- 1.88 N, p = 0.14). CONCLUSIONS: Keratinocyte growth factor-2 did not enhance esophagogastric anastomotic wound healing in this model. However, the dosage and route of administration of KGF-2 may have been sub-optimal for promotion of anastomotic healing. Further studies with this new growth factor are needed. PMID- 12774455 TI - Positive washing cytology in patients with pancreatic cancer indicates a contraindication of pancreatectomy. AB - BACKGROUND: Although peritoneal washing cytology has been widely used in the field of gynecology, it has not been performed so frequently in patients with pancreatic cancer. Only a few papers have reported surgical implications of peritoneal washing cytology. MATERIALS AND METHODS: We reviewed results of peritoneal washing cytology performed immediately after laparotomy in 50 Japanese patients with pancreatic cancer. The 50 patients were divided into two groups according to the results of cytology. Clinicopathological findings were compared between these two groups. RESULTS: Cytology of peritoneal washing was positive in 13 (26%) of the 50 patients. Nine of the 13 patients had no visible peritoneal dissemination. There were no significant differences regarding the age, tumor size and serum level of the carcinoembryonic antigen. Moderately to poorly differentiated adenocarcinoma and perineural invasion (ne3) and lymphatic permeation (ly3) of marked degree were more frequent in the positive group than in the negative group (p < 0.01). The survival curve of the 37 patients with negative cytology was significantly better than that of the 13 with positive cytology (p < 0.01). Four of 13 patients with positive cytology underwent a potentially curative resection but died within 12 months. Three of these four patients died of peritonitis carcinomatosa with massive malignant ascites and one died of local recurrence. CONCLUSIONS: Positive peritoneal washing cytology is not rare in patients with pancreatic cancer and indicates a contraindication of pancreatectomy. PMID- 12774456 TI - Predictors of in-hospital mortality following operative management of hip fractures. AB - OBJECTIVE: To identify predictors of in-hospital mortality and hospital stay following hip fractures. DESIGN: Retrospective cohort study of 185 consecutive patients. SETTING: Tertiary Care University Hospital. PARTICIPANTS: Individuals requiring operative treatment of a proximal femoral fracture excluding those individuals < 50 years old, with femoral head or subtrochanteric fractures, and significant co-morbidity. OUTCOMES: In-hospital mortality and length of hospital stay (days). RESULTS: 116 patients met the inclusion criteria. Predictors of in hospital mortality from logistic regression analysis included male gender (odds ratio with 95% CI: 5.5, 1.5-20.5), admission from a long term care facility (5.5, 1.4-22.6), age greater than 90 years (4.5, 0.9-22.1), and living at home with support (0.2, 0.03-0.9). Predictors of hospital stay from multivariate regression analysis in order of magnitude included presence of a post-operative complication (odds ratio with 95% CI: 14.1, 4.7-44), living at home with support (3.4, 1.3 8.9) and older age (> 85 years) (2.7, 1.0-7.3). Moreover, confusion, urinary tract infections and decubitus ulcers accounted for greater than 50% of all complications encountered. A trend between the number of positive predictors and length of hospital stay was observed. CONCLUSIONS: Elderly individuals admitted from a long term care facility are at high risk of mortality following operative fixation of hip fractures. Early recognition and aggressive management of post operative complications such as confusion, urinary tract infections and decubitus ulcers through careful patient monitoring may decrease hospital stays in those that survive. PMID- 12774457 TI - Blood flow in the pancreatic segment before and after transplantation. AB - BACKGROUND: Blood flow in the pancreas before and after transplantation has not been studied sufficiently. Blood flow in the pancreatic transplant as a function of type vascular anastomosis used has not been explored. AIM: The objective of our study was to study blood flow in the intact pancreas prior to harvesting and at different time periods after transplantation. Two different types of vascular anastomosis were used for segmental transplantation of the pancreas. Blood flow in the transplanted pancreas was compared in this two experimental groups. METHODS: Study was conducted on 61 mongrel dogs. Autotransplantation of a pancreatic segment was performed to the left iliac vessels. There were two randomized experimental groups. In the first group (26 animals) traditional vascular anastomosis was used to revascularize pancreatic graft. Proximal end of splenic artery was anastomosed to iliac artery in "end to side" manner. Portal end of splenic vein was anastomosed to iliac vein in "end to side" manner as well. Splenic ends of splenic artery and vein were ligated. In the second group (35 animals) double arterial double venous anastomosis was used. For this purpose, after completing traditional vascular anastomosis, second arterial anastomosis was created distal to the first one between distal end of splenic artery and iliac artery in "end to side" fashion. Second venous anastomosis was performed distal to the first one between splenic end of splenic vein and iliac vein in "end to side" fashion. Blood flow in the pancreatic segment was measured using direct blood flow measurement method and electromagnetic flowmetry. After transplantation of pancreatic segment, blood flow was measured in the pancreatic graft 3 min, 1 h, 1 and 6 months following transplantation. RESULTS: Blood flow in the pancreas before harvesting was 0.69 +/- 0.01 ml/min/g. Blood flow in the pancreatic transplant in traditional vascular anastomosis group was higher than blood flow in the normal pancreas (1.13 +/- 0.05 ml/min/g after one month and 1.00 +/- 0.05 ml/min/g after six months (p < 0.01)). Blood flow in the double arterial double venous anastomosis group was not statistically significantly different from blood flow in the normal pancreas at 1 and 6 months after transplantation (0.70 +/- 0.02 and 0.67 +/- 0.02 ml/min/g accordingly). CONCLUSION: Double arterial double venous anastomosis provided more physiological blood flow to the graft, than traditional vascular anastomosis. PMID- 12774459 TI - Diagnostic laparoscopy for suspected appendicitis: only useful in young women. AB - The clinical diagnosis of acute appendicitis is inaccurate and can be associated with a negative appendicectomy rate in excess of 40%. The latter has an attendant, albeit low, potential for increased morbidity and mortality, particularly related to infection. In this two year prospective audit of 282 patients presenting with the symptoms and signs of acute appendicitis, preliminary diagnostic laparoscopy (PDL) was performed on 145 patients with the remainder (137) proceeding directly to open appendicectomy. In the PDL group, appendicectomy was undertaken when there was clear clinical evidence of inflammation or when the appendix was not visualised. The negative appendicectomy rate was similar in both groups (21.6% vs. 21.2%) with a male predominance in the PDL group. Whereas women of childbearing age obtained most benefit from PDL with avoidance of unnecessary open operation, the results of our audit showed little benefit for men. PMID- 12774458 TI - Interleukin-10 inhibits inducible nitric oxide synthase in an animal model of necrotizing enterocolitis. AB - BACKGROUND: Nitric oxide (NO) and its role in surgical inflammation are well documented; demonstrating the role of NO in necrotizing enterocolitis (NEC) and ways in which it may be suppressed may provide avenues for immune modulation in the treatment of NEC. AIMS: We sought to demonstrate an increase in inducible nitric oxide synthase (iNOS) mRNA and nitric oxide in an experimental model of necrotizing enterocolitis. In addition, we hypothesized that interleukin-10 (IL 10) would attenuate this response. METHODS: Newborn rats were treated with 25 microliters intraperitoneal IL-10 or vehicle prior to laparotomy, 1 h superior mesenteric artery (SMA) occlusion, 50 micrograms/kg intraluminal platelet activating factor administration, and SMA reperfusion. iNOS mRNA and nitric oxide levels were measured in the liver, small bowel, and serum and compared using Student's t-test. RESULTS: Small bowel iNOS mRNA increased after NEC induction from 0.058 +/- 0.02 to 0.144 +/- 0.05 relative intensity units (RIU) at 2 h (p < 0.01) and from 0 to 0.09 +/- 0.02 RIU at 6 h (p < 0.03). Liver mRNA increased from 0.026 +/- 0.002 to 0.485 +/- 0.09 RIU (p < 0.002) and from 0 to 0.069 +/- 0.02 RIU (p < 0.0001) at 2 and 6 h, respectively. Serum nitric oxide increased in NEC induced animals at 2 h from 28.04 +/- 10.5 to 45.18 +/- 6.8 microM (p < 0.001). IL-10 suppressed iNOS mRNA and nitric oxide expression at 2 h in small bowel, liver, and serum by 60%, 89%, and 11%, respectively. CONCLUSIONS: IL-10 decreases iNOS mRNA response in experimental NEC. This down-regulation may be an avenue for anti-inflammatory intervention in NEC. PMID- 12774460 TI - Laparoscopic versus conventional Nissen fundoplication: the influence on plasma insulin-like growth factor-I and insulin-like growth factor binding protein-3. AB - BACKGROUND: Conventional surgery is known to influence postoperative insulin-like growth factor-I (IGF-I) levels and to increase protein breakdown. Recent animal data described that laparoscopic surgery, in contrast to conventional surgery, maintains postoperative IGF-I levels, suggesting a reduced postoperative catabolic response. AIM: The aim of this study was to demonstrate differences in the postoperative catabolic response between laparoscopic and conventional surgery. METHODS: Seventeen patients scheduled for Nissen fundoplication were randomly assigned to undergo either a laparoscopic or a conventional procedure. The catabolic response was assessed by measuring total IGF-I and insulin-like growth factor binding protein-3 (IGFBP-3). RESULTS: IGF-I levels decreased significantly 4 days after both types of surgery. IGFBP-3 decreased more rapidly after conventional surgery than after the laparoscopic technique; however, the differences between the groups were not significant. CONCLUSION: In contrast to animal data, no differences in plasma IGF-I or IGFBP-3 were seen in patients undergoing laparoscopic or conventional Nissen fundoplication. PMID- 12774461 TI - Esophageal function studies in the management of gastroesophageal reflux disease. AB - BACKGROUND: Esophageal function testing, i.e. esophageal manometry and 24-h pH monitoring, are usually carried out to diagnose gastroesophageal reflux disease (GERD) in patients with atypical symptoms, when there is no evidence of esophagitis at endoscopy, or following previous unsuccessful surgery. Additionally, these studies should be considered mandatory before surgery to confirm the diagnosis and to tailor the procedure to the motility pattern of the individual patient. AIM: The aim of this study was to assess the role of esophageal function studies in the management of patients with GERD. METHODS: Patients with a mechanically defective lower esophageal sphincter (LES) and abnormal esophageal acid exposure proven at 24-h pH monitoring were considered for surgery. A 360 degrees fundoplication (Nissen) was performed in patients with good esophageal motility, whereas a partial 180 degrees fundoplication (Toupet) in patients with a defective motility. Five hundred and eighty-six patients were referred to our laboratory for symptoms suggestive of GERD between November 1992 and April 1999. RESULTS: Twenty-four hour esophageal pH monitoring was positive in 65.5% of these patients; manometry showed a defective lower esophageal sphincter and a defective esophageal body motility in 57.8% and 21.7% respectively. One hundred and two patients underwent a total fundoplication, and 43 patients a partial wrap. At a mean follow-up of 36 months (range 5-69) the actuarial success rate in the control of reflux was 90%. Three (2.9%) patients undergoing a Nissen fundoplication complained of persistent dysphagia; two of them had preoperative esophageal body dysfunction. CONCLUSION: Esophageal function studies allow proper selection of patients for surgery and guide in the choice of the antireflux operation. PMID- 12774462 TI - Symptomatic and asymptomatic carotid artery lesions in peripheral vascular disease: a prospective study. AB - PURPOSE: To assess the incidence and severity of internal carotid artery lesions in patients who were referred to a single institution with peripheral obliterative atherosclerotic disease (POAD) as their chief complaint, in order to identify symptomatic and asymptomatic subjects at risk of stroke who would benefit from surgery. MATERIALS AND METHODS: Consecutive patients with clinically and Doppler scan-proven POAD underwent prospective screening for the presence of carotid symptomatic and asymptomatic atherosclerotic lesions with duplex ultrasound scan (DUS). Associated risk factors for atherosclerosis, i.e. age, sex, diabetes, hypertension, hyperlipidemia, history of smoking and history of coronary artery disease were assessed and recorded. Univariate analysis was performed to correlate the risk factors with the presence of POAD and carotid lesions. RESULTS: Three-hundred and twelve consecutive patients were evaluated over an 18-month period. A 30% or greater carotid stenosis was detected by DUS in 189 patients (19 bilateral); 57 (30%) of these had neurologic symptoms, 19 of whom had a 70-99% carotid lesion, whereas 65 of the 132 asymptomatic patients had 60-99% carotid stenosis. Univariate analysis revealed no differences in the symptomatic and asymptomatic groups between patients who were potential candidates for carotid endarterectomy according to the North American Symptomatic Carotid Endarterectomy Trial and the Asymptomatic Carotid Atherosclerotic Study Group guidelines, respectively, and those who were not as regards associated atherosclerotic risk factors and severity of POAD. All but 2 (n = 17) symptomatic and 18 asymptomatic patients underwent carotid endarterectomy based on DUS criteria alone. The perioperative stroke risk and mortality rates were 0%. CONCLUSIONS: Routine carotid DUS screening of 312 consecutive patients with POAD revealed that 19 patients with symptoms and 65 (5 bilateral) without symptoms would benefit from surgery, representing 27% of the series considered. PMID- 12774463 TI - [Summing up of functioning of the system of compulsory medical insurance in the Russian Federation and prospects for its formation in Ukraine]. AB - Submitted in the paper are data obtained as a result of analyzing and pooling information now available about positive and negative sequels of introduction in the Russian Federation of a compulsory medical insurance. Emphasis is placed on the relevance of, and need for, taking account of practical experience gained by the Russian Federation in deciding on the question about time and expediency of the start of a similar reform in Ukraine and on the importance of use of innovational approaches and technology at the stage of its planning. PMID- 12774464 TI - [Derivatives of cholecalciferol in the treatment of psoriasis]. AB - The review of the literature is devoted to derivatives of calciferol (vitamin D3), representatives of a new group of antiproliferative drugs employed in psoriasis, ichthyosis, rheumatism and some neoplastic processes. Results are submitted of histo- and biochemical, physiological, and clinical trials of the synthetic derivative of vitamin D calcipotriol, a possibility of its combination with physiotheraputic modes of treatment is reported together with recently identified side effects. Novel analogues of the drug are listed, with their merits and demerits pointed out. The authors conclude that further studies are warranted into abnormalities of metabolism of calcium in psoriasis to develop efficacious modes of correction of dermatosis involving the use of representatives of the given group of medicinal agents. PMID- 12774465 TI - [Mosaicism of the upper respiratory tract epithelium in liquidators of the Chernobyl accident]. AB - On the basis of findings from the structural analysis of 21 nasal mucosa biopsy specimens of patients with chronic rhinitis including 12 participants in liquidation of the consequences of the Chernobyl accident using morphometric methods, signs of morphofunctional insufficiency of the nasal epithelium are verified. Mechanisms of this phenomenon development are discussed. PMID- 12774466 TI - [Indices of humoral and cellular immunity in patients with chronic maxillary sinusitis after the Chernobyl accident]. AB - An evaluation was done of immunological parameters in the post accident period in 41 examined person. Of these, 26 subjects had chronic maxillary sinusitis, 15 were healthy persons (control group). In patients with chronic maxillary sinusitis, the state of the T-system of immunity has been shown to be characterised by a drop in the total count of T-lymphocytes, T-helpers, T suppresors. Parameters for humoral immunity are characterised by elevation of the peripheral blood levels of B-lymphocytes, prominent disimmunoglobulinaemia- significant reduction of class G immunoglobulins and macroglobulins. The T-cells helpers ratio is significantly reduced both in group I and group II. Reduction has been ascertained in the ratio of those T-cells-suppressors blocking antibody formation by B-lymphocytes. In patients after the Chernobyl accident, there was noted a significant decline in the functional activity of macrophages. PMID- 12774468 TI - [Regulation of tonus of the autonomic nervous system in patients with frequently relapsing atrial fibrillation]. AB - An evaluation was carried out of variability of the heart's rhythm in 269 patients with ischemic heart disease and/or hypertensive disease, their age ranging between 43 to 82 years, complicated by frequently relapsing atrial fibrillation. It is shown that in those patients presenting with frequent recurrences of arrhythmia over the course of a 24-hour period, there comes to be a considerable decline in the power of total variability of cardiac rhythm, and all spectral components and a shift in the sympathetic/parasympathetic system balance in favour of the sympathetic one. Analyzed the study were particular features of regulation of tension of the vegetative nervous system in different clinical forms of atrial fibrillation. Patients with frequently recurring atrial fibrillation have been shown to constitute rather a miscellaneous group as to patterns of the cardiac rhythm variability, which fact is to be taken account of in conducting prophylactic antiarrhythmic therapy of recurrencies of atrial fibrillation. PMID- 12774467 TI - [Hemodynamics and cell membrane effects of angiotensin II receptor antagonists in treatment of hypertension]. AB - Strain on calcium-dependent mechanisms in essential hypertension (EH) is in activation of the calcium pump, structural-and-functional changes in Ca2+ ATPase and activation of proteinase C. In EH patients presenting with a high sensitivity to angiotensin II, there comes to be frequently seen an insufficient effectiveness of antagonists of receptors to angiotensin II. PMID- 12774470 TI - [Pathogenetic mechanisms of anemia in children with celiac disease]. AB - Pathogenetic mechanisms of anemia development were studied in children with celiac disease to develop guiding rules for administration of complex therapy. It has been found out that celiac disease in children is accompanied by a high incidence of grave forms of anemia that adversely affect the course of the underlying condition. In anemic children with celiac disease, manifest pathophysiologic and pathobiochemical alterations in the hemoglobin and iron status are most common, with anemia of hyperregeneratory character being more prevalent. In children with celiac disease, the complex of therapy of anemia that do not respond to the instituted treatment options should be supplemented with protein preparations or activators of protein metabolism and with medicinal agents capable of correcting intestinal microflora. PMID- 12774469 TI - [Lipid peroxidation and enzymes of the antioxidant defense system in patients with bronchial asthma]. AB - In 71 patient with bronchial asthma (BA), lipid peroxidation (LPO) was studied together with the condition of the antioxidant system (AOS) as related to the degree of severity of the illness, with results evaluated of conventional therapy versus complex treatment involving the use of cytochrome C. Established in the above series was intensification of LPO in the presence of inhibition of activity of SOD enzymes and catalase, their degree dependent on severity of the condition. Unlike basic therapy, employment of cytochrome C has been found to be associated with a more complete restoration of balance in the LPO/AOS system, especially so in steroid-independent form of BA. PMID- 12774471 TI - [Sulfonylurea receptors and their interaction with glimepiride]. AB - In the review, data are presented on glymepiride (amaril), a 3-d generation sulphanylamide. The drug interacts with receptors to sulphonylurea--SUR X, that are found only in B-cells of the pancreas, which fact has been shown to be associated with a higher secretion of insulin, an increased sensitivity of peripheral tissues to this hormone and with insulin-like extrapancreatic effects as well. Amaril does not affect SUR of the myocardium, myocytes of cerebral vessels and nervous cells. The use of the above drug combined with metformin, insulin, repaglynide, and nateglynide has been shown to be an optimum treatment option, however in the latter case, hypoglycemic states may come to be seen more frequently along with a rapid exhaustion of B-cells. In view of the fact that in many instances, type 2 diabetes mellitus runs its course concurrently with other illness, it is necessary that interaction of amaril with other drugs be taken into account. PMID- 12774473 TI - [Leech therapy of patients with reactive arthritis]. AB - Results are submitted of bdellotomy in patients with reactive arthritis. Shown in the above patients were palliative and and anti-inflammatory effects of bdellometers. Bdellotomy has proved to be efficient treatment option in a major proportion of patients. Bdellotomy will, we believe, come to be a valuable adjunct in the physician's therapeutic arsenal, that can be used not only in the treatment of patients with rheumatic disorders but by family doctors as well and in other general practice settings. PMID- 12774472 TI - [The incidence and antibiotic resistance of coagulase negative staphylococci isolated in pregnant women and newborns]. AB - Puerparae and newborns infants display an ever-increasing colonization with not only opportunistic enterobacteria but also coagulase-negative staphylococci that have a multiple resistance to antibiotics. In the study, the identification has been carried out of staphylococci, with their antibioticoresistance defined. PMID- 12774474 TI - [Clinical manifestations of secondary pneumonia in patients of general surgical departments]. AB - Analysis of records of surgical patients and protocols of anatomicopathological investigations in autopsy tissues with changes in the lungs characteristic of pneumonia suggest to us that secondary pneumonia comes up to 38.5% of all post mortem examinations. In 9 percent of the deceased, secondary pneumonia appeared to be a direct cause of death. The frequency rate of pneumonia in operated patients ranged between 0.3-0.5% and 68%, this concerning largely aged and senile patients and also those patients with polytraumata, in surgical interventions in organs of the thorax, abdominal cavity, in full-thickness burns. A considerable number (30%) of diagnostic errors of pneumonia in surgical departments is due to a complicated character and extreme diversity of clinical manifestations of the underlying medical condition or polytraumata. PMID- 12774475 TI - [Diagnosis and classification of thoracal endometriosis]. AB - Endometriosis is a grave pathological process that comes to be encountered in young women of the reproductive age, and it is not infrequently that it terminates in disability. Apart from genital organs, other organs may be affected by the disease, including those of the thorax. But in proportion of instances the process escapes early diagnosis. In the author's opinion, the reason for the above-said is absence of classification, in view of which fact a classification is suggested that is supposed to advance diagnosis. PMID- 12774476 TI - [The incidence, features of the course, and treatment of arterial hypertension in patients with prostate hyperplasia]. AB - In 144 middle-aged and elderly male subjects with benign prostate hyperplasia, a study was made into incidence and course features of concomitant arterial hypertension (AH). AH has been proved to be encountered in 144 percent of patients with benign hyperplasia of the prostate. It has been ascertained that in 84% of patients, irritative and obstructive symptoms related to benign hyperplasia of the prostate adversely affect the course of concomitant arterial hypertension. Striking reduction of manifestations of benign prostate hyperplasia related irritative and obstructive symptoms using cardura, a selective alpha adrenoblocker, has come to significantly enhance efficiency of treatment of concurrent AH. PMID- 12774478 TI - [Complex treatment of the pro- and antioxidant system disorders in patients with psoriasis]. AB - The consition was studied of the pro- and antioxidant system in psoriatic patients. The groups of the examinees comprised 44 patients in the progressive stage of dermatosis and 16 patients in its stationary stage. Examined in the study was also a group of control subjects (16 and 10 patients respectively) and 15 healthy volunteers. The group of those patients presenting with a progressive process demonstrated increased indices for both lipid peroxidation and antioxidant defence. Examination of patients in the stationary stage of psoriasis revealed a significant intensification of processes of lipid peroxidation (rise in malonic dialdehyde, in dienic conjugates, and enhanced chemiluminiscence) associated with a decline in the activity of processes of the antioxidant defence. Expediency is proved of differentiated employment of antioxidant preparations depending on the stage of psoriasis. PMID- 12774477 TI - [A new approach to the treatment of sleep disorders in patients with cerebrovascular diseases]. AB - A positive result has been obtained in treating sleep disorders in patients with chronic vascular affections of the brain with using the drugs donormyl and sonnate-KM [symbol: see text]. Donormyl is recommended for use in sleep disturbances (not very prolonged and not profound insomnia) in a 10-day therapeutic course as a first-choice drug. PMID- 12774479 TI - [The role of uric acid in effects of balneotherapy on the phagocytic link of immunity and unspecific defense at the Truskavets health resort]. AB - Revealed in the wake of balneotherapy at the Truskavets health-resort were two opposite types of changes--those in the level of uriacidemia and in the content of theophylline-sensitive lymphocytes that are probably caused by central and peripheral effects of uric acid. In this setting, parameters characterizing the phagocytic link of immunity and unspecific defense are noted to change unidirectionally, in other words, they remain unaffected by changes in uriacidemia. PMID- 12774480 TI - [Modern methods of diagnosis of giant-cell tumor of bone]. AB - As many as 170 patients with giant cell tumour of the bone were examined in the Institute of Oncology department of tumours of the locomotorium. The diagnosis of giant cell tumour of the bone was made with a comprehensive approach to the patients pursued to include roentgenography, computerized tomography, biopsy involving histological study of the specimens taken. PMID- 12774481 TI - [Medico-genetic counselling and surgical correction of different forms of facial clefts in children]. AB - In the paper, consideration is given to the necessity of a medicogenetic counselling of children with different forms of facial cleft as an important stage in the therapeutic policy permitting carrying out differential diagnosis with a syndromological approach, verifying the diagnose made, calculating a returned genetic risk for the family, and formulating a policy of administering prophylaxis of birth of a child with cleft lip, alveolar process, hard and soft palate. Special stress is laid on staging of surgical treatment of children presenting with the developmental defect under consideration, the need for colleagues in multiple disciplines, such as maxillofacial surgeon, orthodontist, logopedist, otolaryngologist, pediatrician, genetician, psychologist, which measures will, we believe, help in securing a successful rehabilitation and social adaptation of a child. PMID- 12774482 TI - [Prophylaxis of pyogenic complications in total replacement of large joints]. AB - Results are studied of management of 378 patients having undergone total knee and hip arthroplasty over the period 1995-2001. Surgical interventions were undertaken in those patients presenting with osteoarthrosis deformans, rheumatoid arthritis, for complicated posttraumatic fractures, pathological fractures secondary to an oncological process. Pyo-inflammatory complications which necessitated removal of endoprothesis developed in nine (2.32%) patients. A strict keeping to the system of the purulent complications prophylaxis in total arthroplasty is an important component of a successful management of those patients presenting with grave forms of affections of the great joints. PMID- 12774483 TI - [Assessment of effectiveness of livolin forte in the treatment of patient with steatohepatitis]. AB - The article analyzes efficacy of the drug livolin in the treatment of patients with fatty hepatosis. It is shown that livolin promotes a rapid arresting of clinical symptomatology, normalization of biochemical indices, makes for improvement of physical-and-chemical properties of the bile. PMID- 12774484 TI - [Comparative results of therapy with amizon and reaferon of patients with chronic viral hepatitis B and C and kidney complications]. AB - A comparative clinical-and-laboratory investigation was undertaken designed to study efficiency of employment of reaferon and amison in 30 patients with viral hepatitis B and 30 patients with viral hepatitis C and renal lesions. Amison has been shown to significantly reduce or dispel dispeptic events in the above patient populations, with no serious ill effects noted. Amison has proved to be endowed with the ability to exert apparent antiinflammatory, interferonogenic, immunomodulating, and hepatoprotecting effects, and it is much cheaper compared to reaferon. PMID- 12774485 TI - [Comparative economic and clinical effectiveness of setegis and camiren in therapy of chronic prostatitis]. AB - The drugs setegis 2 mg, camiren 2 mg, were used in the investigation designed to study economic and clinical effectiveness of employment of alpha-blockers for treating chronic prostatitis, with the expediency of the above therapeutic modality having been corraborated by the findings secured. Objective control of the treatments administered necessitates studying the time-related changes in indices for the amount of the residual urine and volume of the prostate. Economic expediency has been shown of use of camiren 2 mg, an alpha-blocker, of the KRKA firm, which fact was validated by a lower cost of the drug at the retail market of Ukraine compared to setegis 2 mg. PMID- 12774486 TI - [The method of preservation of cadavers in a home setting]. AB - Based on the results of preservation of 871 dead bodies in home settings a composition of balsamic solution is proposed together with a mode of its administration to preserve corpses in a better way for a short time (within several days). PMID- 12774487 TI - [The role of labour conditions in the development of osteo-muscular system disorders in workers of the iron-ore industry]. AB - As a result of the investigation conducted, differences have been revealed of effects of the studied factors of labour conditions and its quantitative values on development of disorders of the osseomuscular system depending on the character of labour. In workers of basic occupations in iron-ore mines, the development of radiculopathy qualified as an occupational problem, has been shown to be determined by exposure to a complex of studied adverse factors of the occupational environment and labour process by 35% (very high degree of the effect exerted), including local vibration--by 14%, time of keeping a required working posture standing--by 7.2%, air temperature difference--by 7.4% (high and moderate degree of the effect exerted). The above adverse factors are to be considered in sanitary-and-hygienic characterization of the working place and taken account of in attempts to ascertain relatedness of the illness in question to the worker's occupation and in implementing measures designed to administer primary and secondary prophylaxis. In those workers engaged in occupations of the sensomotor kind operator's work, development of spinal osteochondrosis is determined by 25.4% by action of a complex of the studied factors of labour conditions; of this percentage, 22.4% have been shown to be caused by a number of working (operating) movements during the shift (very strong impact), which fact necessitates qualifying the condition as an occupational disease and suggests increase in the role of improvement in labour conditions in its prophylaxis. PMID- 12774488 TI - [Significance of immunoenzyme analysis in early diagnosis of dust bronchitis]. AB - Measurement of levels of antibodies to a causative agent of tuberculosis with the aid of the enzyme immunoassay can serve as one of methods for early diagnosis of infectious- and dust diseases. Patients with clinically established diagnosis of dust bronchitis, latent silicotuberculosis and manifest silicotuberculosis demonstrated a significantly higher level of antituberculous antibodies in blood plasma as compared to controls. Establishment of the above fact permits regarding a major proportion of dust bronchitis as a silicotuberculous process. PMID- 12774489 TI - [A social and hygienic assessment of the population health in the context of its reproductive formation]. AB - An analysis of medicodemographic processes has been done together with that of certain indices for case rate in the urban and rural populations residing in the Precarpathian Region territories over a 10-year period. Revealed by the above analysis were regional depopulational patterns child mortality patterns, trends towards variations in case rates. A conclusion has been reached as to cause-and effect depopulation processes, features of variations in case rate, and abnormalities in reproductive populational formation as well. PMID- 12774490 TI - [Methodological approach to the creation of theory of medical care of patients with proximal hip fracture]. AB - In the present article, clinically effective principles of treatment of fractures, used in creation of a doctrine of service delivery to patients presenting with proximal hip fractures (PHF), are pooled and compared. The place of insurance medicine in the delivery of relevant services to PHF patients is determined together with chief items in realization thereof, such as proper organization of processes of care of public health, provision of specialized care for patients in the above nozological group, material support, personified data base. PMID- 12774491 TI - [Some aspects of disability in patients with diabetes mellitus pa]. AB - In the article, results are submitted of the study of disability of those persons presenting with diabetes mellitus (DM) in Tashkent over the period 1997-1998. Among DM subjects referrable to the examination, there prevails type 2 DM (13.9 +/- 1.6) and (10.6 +/- 0.7) years in duration in 1997 and 1998 respectively. Average DM subject age was (52.7 +/- 1.0) in 1997 and (49.7 +/- 0.8) in 1998 at the time the examined person was declared an invalid. Invalidism is noted to be on the increase (141%). At examination, more than 80 percent of persons presenting with DM are declared disabled. PMID- 12774492 TI - [Problems of the birth rate of population and scientific-practical approaches to their solution]. AB - Data are submitted to the effect that the population birth-rate is a national, political, and social problem rather than medical one only. An account is given of those factors affecting the level of birth-rate and health of newborn infants at the present stage of development of the society. Chief requirements the programme is to comply with are specified. Carrying out the projects suggested will, we believe, help in putting an end to further aggravation of the crisis. PMID- 12774493 TI - [Seventy-fifth anniversary of the D. K. Zabolotnyi Institute of Microbiology and Virology of the National Academy of Sciences of Ukraine (Stages of the great path)]. PMID- 12774494 TI - [Some comments about Valerii Veniaminovych Smirnov--chief, teacher, colleague and comrade]. PMID- 12774495 TI - [Danylo Kyrylovych Zabolotnyi--founder of the Institute of Microbiology and Virology of the National Academy of Sciences of Ukraine--scientist and citizen]. PMID- 12774496 TI - [Viktor Hryhorovych Drobot'ko: a life deserving immortality]. PMID- 12774497 TI - [Department of Antibiotics--its history and the present]. AB - The paper on the occasion of the 75th anniversary of the Institute of Microbiology and Virology of the NAS of Ukraine is dedicated to scientific activity of one of its oldest subdivisions: the Department of Antibiotics. The main trends of scientific research and their most important results for the whole period of the Department existence are presented. PMID- 12774498 TI - [About the past and present of the Department of General and Soil Microbiology and ... something else]. AB - A historical essay of creative path of collective of the Department of General and Soil Microbiology, founded in 1928, is presented. Two research schools have been formed for the past years: ecophysiology of soil microflora and geochemical activity of microorganisms. L. I. Rubenchik stood at the sources of these schools. A problem of soil biology and microorganisms interrelations with higher plants occupies a considerable place in the Department research. Separate groups, microorganisms and microbe cenoses are studied as a whole as the complex self regulated system. The investigations of microbe corrosion of metals initiated by a necessity to solve urgent problems of Kyiv metro have been formed as the priority trend. Agents and mechanisms of biological damages of the overground, underground and underwater structures are studied and the grounds of anticorrosion measures are developed within the framework of these trends. PMID- 12774499 TI - [Role of Semen Mykytovych Moskovets' in the foundation and development of plant virology at the Institute of Microbiology and Virology of the National Academy of Sciences of Ukraine (1960-1971)]. AB - The historical events in the Zabolotny Institute of Microbiology and Virology of the NAS of Ukraine that were connected with professor's S.M. Moskovets work activity as director and organizer of plant virology investigations are analyzed. PMID- 12774500 TI - [Seventy years of research in experimental mycology at the Institute of Microbiology and Virology of the National Academy of Sciences of Ukraine]. AB - The history of development of mycology at the D.K. Zabolotny Institute of Microbiology and Virology of the NAS of Ukraine and main achievements of the Department of Physiology and Taxonomy of Micromycetes during 1933-2003 were described in this publication. PMID- 12774501 TI - [Dynamics of research on phytopathogenic bacteria at the Institute of Microbiology and Virology of the National Academy of Sciences of Ukraine]. AB - The basic trends of the work of the Department of Phytopathogenic Bacteria from the beginning of its existence in the system of IMV of the NAS of Ukraine (1934) have been analyzed. The stages of personnel training, the Department relations with other departments of IMV and various institutions, investigation of plant bacterioses, regularities of appearance and development of the infection process, methods of diseases control have been distinguished. Special attention was given to phytopathogenic bacteria: ecology, morphology, antigens and their structure. Poliobiotrophy, heterogeneity of pathogens population and its relation with pathogenicity have been investigated. Many-sided relations of pathogens with epiphytes and endophytes have been revealed. Bacteriophages of phytopathogenic bacteria have been studied. PMID- 12774502 TI - [Department of Biochemistry of Microorganisms--start of the path (1951-1973)]. AB - Creation of the Department of Biochemistry of Microorganisms at the Institute of Microbiology and Virology of the Academy of Sciences of Ukrainian SSR in the 30's of the last century was determined by a necessity of profound investigation of vital activity biochemism of microorganisms from various systematic groups which were studied in microbiological department of the Institute. Such complexity can explain certain diversity of the Department research at initial stages of its existence. The research of saccharose transformation into dextran Leuconostoc mesenteroides, when production solutions become slingy at sugar-refinaries, was one of the first most significant works of the Department. The enzyme saccharose glycosyl-transferase performing this process was described for the first time. A cycle of works on the study of enzymes splitting lactose in milk under the effect of Streptococcus lactis has been carried out. Complex investigation of a number of proteins, polysaccharides, enzymes in enterobacteria has shown that the blocking of the enzyme aldolase is one of the reasons of alkali formation. A method has been developed for isolation of arenarin, antibiotic of plant origin, from sandy everlasting, the nature of its acting basis has been established. Nufarin, an active antibiotic, was isolated from the roots of white water lily when studying nitrogen fixation processes, special attention was given to interaction of hydrogenase and enzymes, taking part in nitrogen fixation, to the effect of ATP on these processes, ways of its synthesis, localization of ATPase in the cell membranes. Works on the study of lypopolysaccharides and polysaccharides of Gram-negative enterobacteria, bacteria of Pseudomonas genus were started with the purpose to use the obtained data to specify systematic propositions of the investigated microorganisms. Further on these works became the basis of thematic department. There are numerous reviews dedicated to their development. PMID- 12774503 TI - [Formation of research on viruses of humans, animals, and microorganisms at the Institute of Microbiology and Virology of the National Academy of Sciences of Ukraine and its present development]. AB - The 40-year history of virology development at the D.K. Zabolotny Institute of Microbiology and Virology of the NAS of Ukraine and the basic success achieved in the study of human, insect viruses, phages of phytopathogenic bacteria have been analyzed. PMID- 12774504 TI - [Thirty-five years of the Department of Biology of Gas-oxidizing Microorganisms of the D.K. Zabolotnyi Institute of Microbiology and Virology of the Academy of Sciences of Ukraine]. AB - The basic trends of the department scientific activity are presented. The priority results of investigations of methaneoxidizing bacteria biology are given. The biothechnological development of the department (obtaining of products of microbial synthesis: protein, polysaccharides, antialcoholic and antinarcotic preparations) are considered in detail. Possible areas of their application are presented. PMID- 12774505 TI - [Development of cyanobacterial phages at the Institute of Microbiology and Virology of the National Academy of Sciences of Ukraine (History and perspectives)]. AB - The paper deals with the basic trends of fundamental investigations of the Department of Algae Viruses in the field of cyanophagia-ecology, biological and physico-chemical properties of cyanophages as well as interrelation with the host cells. Such problems as a possibility to use the system cyanophage-cyanobacteria as the experimental model for development of the unified functional model of productive infection, efficient methods of prophylaxis and therapy of virus infections as well as the solution of various biotechnological problems are discussed. PMID- 12774506 TI - [Mycotoxins: past, present and future]. AB - The paper deals with historical information concerning the investigation of mycotoxins and mycotoxicoses at the Institute of Microbiology and Virology of the NAS of Ukraine, main results of the works fulfilled for the last 25 years and possible trends of further investigations. PMID- 12774507 TI - [Taxonomy, ecology, and physiological and biochemical characteristics of industrially important microorganisms]. AB - The work is dedicated to the investigations which were carried out in the Department of Physiology of Industrial Microorganisms in the recent years. Fundamental results of investigations dedicated to development of systematics of nocardio- and coryne-like bacteria, lactic acid bacteria and yeast; to the study of their distribution in nature, physiological-and-biochemical peculiarities. Considerable attention is paid to growth physiology of lactic acid bacteria and yeast, to lectin-forming microorganisms, to the study of fungi shaping under deep cultivation, as well as to microorganisms, taking part in the environment purification from heavy metals and oil products. Drugs created for treatment and prophylaxis of cattle diseases, for fodder siloing; new lactic acid preparations of functional direction, technological bases of sewage treatment for oil products and heavy metals are described. PMID- 12774508 TI - [Streptomycetes--producers of polyketide antibiotics]. AB - The review is dedicated to biosynthesis of polyketides compounds by streptomycetes. It contains new data about structure of streptomycete chromosome, structure and action mechanism of polyketidesynthases of I and II types and gene clusters determining the biosynthesis of polyketides. A new promising approach is considered--the combinatorial biosynthesis of hybrid "unnatural" antibiotics by means of gene engineering construction of recombinant strains bearing the biosynthetic genes of bacterial aromatic polyketidesynthases. PMID- 12774509 TI - [Current trends in the study of microbial glycopolymers]. AB - The prospects of studying glycopolymers both in theoretical and practical aspects are presented. The paper deals with: 1) different strategies of the struggle with endotoxical shock, caused by lipopolysaccharides of gram-negative bacteria, 2) role of carbohydrates in the design of drugs, including antibiotics, 3) use of the enzyme inhibitors as medicines, 4) possibilities of the use of proteolytic enzymes, collagenase and keratinase, in particular, both for theoretical investigations (the finding of the structure of proteins and peptides), and for their use in practice. PMID- 12774510 TI - [Interferon inducers--from theory to practice]. AB - The authors present up-to-date information concerning different inducers of different interferon types in vivo and in vitro. Interactions between the structure and interferon-inducing activity of high and low molecular weight inducers are in the focus of this paper. There are also a lot of data, concerning different biological properties of interferon-inducing compounds. Different aspects of inducers use as antiviral and immunomodulating drugs are also discussed. PMID- 12774511 TI - [The problem of mollicute resistance to antibiotics]. AB - The problem of disease agents resistance to antibiotics is a general microbiological one, but it was considered in literature with respect to bacteria only. The problem of resistance to antibiotics of mollicutes class representatives has not been touched in generalizing literature. This work is the first attempt to analyze the data from the world literature concerning the appearance of resistance to antibiotics in mollicute species, doing the greatest harm to people, and to present information which would be of consultative value, when choosing antibiotics to treat human mycoplasmoses. PMID- 12774512 TI - [Computer analysis of viral genomic structural organization by sequence scanning]. AB - The results of testing the author's computer program for viral genomic sequences scanning (program Scangene) are presented in the paper. It has been shown on model nucleotide sequences as well as on genomic and predicted protein sequences of tobamo-, potex- and bromoviruses that the program is suitable for the quick revealing of identical, similar and specific sequence elements such as direct, inverted and regular repeats, deletions/insertions, GC/AT-containing sites, fixed intervals between common sites of identical aminoacids in viral coat proteins etc. Characteristics and prospective application of the sequence scanning procedures are discussed. PMID- 12774513 TI - Gamma-tubulins and their functions. AB - gamma-Tubulin is a ubiquitous phylogenetically conserved member of tubulin superfamily. In comparison with alpha beta-tubulin dimers, it is a low abundance protein present within the cells in both various types of microtubule-organizing centers and cytoplasmic protein complexes. gamma-Tubulin small complexes are subunits of the gamma-tubulin ring complex, which is involved in microtubule nucleation and capping of the minus ends of microtubules. In the past years important findings have advanced the understanding of the structure and function of gamma-tubulin ring complexes. Recent evidences suggest that the functions of gamma-tubulin extend beyond microtubule nucleation. PMID- 12774514 TI - Motor proteins and kinesin-based nanoactuatoric devices. AB - Eukaryotic organisms synthesize diverse motor proteins converting chemical into mechanical energy. Among them, both rotary (e.g., ATP synthase) and linear motors are found. Linear motors comprise highly specialized proteins moving along nucleic acid filaments (in the case of e.g., RNA polymerase) or cytoskeletal filaments. The present paper provides a brief overview on cytoskeleton-associated motors (myosins, dyneins, and kinesins) and summarizes results contributing to elaborate a basic configuration for constructing a kinesin-driven motor device, suitable for e.g. a controlled displacement of objects or specific substances over millimetre distances with nanometre precision. PMID- 12774515 TI - [Dynamics and the life cycle of cell microtubules]. AB - In living cells microtubules (MTs) continuously grow and shorten. This feature of MTs was discovered in vitro and named dynamic instability. Comparison of dynamic instability of MTs in vitro and in vivo shows a number of differences. MTs in vivo rapidly grow (up to 20 microns/min), duration of their shortening is small (on average 15-20 s), and pauses are prominent. In different animal cells MTs grow from the centrosome and form a radial array. In such cells growth of MTs is persistent, i.e. undergo without interruptions until plus end of a MT reaches cell margin. Analysis of literature and original data shows that interconvertion between phases of growth, shortening and pause is asymmetric: growth often converts into pause, while shortening always converts into growth without pause. We suggest dynamic instability described near the cell margin in numerous publications results not only from intrinsic properties of MTs, but also because of the external obstacles for their growth. MT behavior in the cells with radial array of long MTs could be treated as dynamic instability with boundary conditions. One boundary is the centrosome responsible for rapid initiation of MT growth. Another boundary is cell margin limiting MT elongation. MT growth occurs with constant mean velocity, and potential duration of growth phase might exceed cell radius. MT shortening is usually smaller than MT length however velocity of shortening increases with time. Random episodes of rapid shortening are sufficient for the exchange of MTs in 10-20 min in the cells not more than 40-50 microns in diameter. Experimental data show that similar rate of exchange of MTs is in the large cells. This is achieved employing another mechanism, namely release of MTs and depolymerization from the minus end. In the minus end pathway time required for the exchange of MTs does not depend on cell radius and is determined primarily by the frequency of releases. Thus a small number of free MTs with metastable minus ends significantly reduce time required for the renovation of the radial MT array. Summarizing all experimental data we suggest the life cycle scheme for the MT in a cell. MT is initiated at the centrosome and grows rapidly until it reaches cell margin. At the margin the plus end oscillates, and finally MT depolimerizes. MT "death" comes from a random catastrophe (shortening from the plus end) in small cells or from release and depolymerization of the minus end in large cells. PMID- 12774516 TI - [Microprotoplasts as an efficient technique for chromosome transfer between incompatible plant species]. AB - The perspectives of restricted plant genome transfer by means of fusion of isolated microprotoplasts containing one or few chromosomes with recipient protoplasts are considered. Efficiency of the usage of antimicrotubular compounds such as phosphoroamidates (amiprophosmethyl and cremart), dinitroanilines (oryzalin) as well as propizamide and microtubule organising center disrupters (griseofulvin and phenylcarbamates) for micronucleation are analyzed. Advantages of the combined usage of antimicrotubular drugs and cytochalasin B as disrupter of actin filaments for enhanced yield of microprotoplasts are considered. Results of subsequent establishment of microprotoplast-mediated chromosome transfer technique is summarized. The examples of successful microprotoplast-mediated transfer of individual chromosomes from a donor potato line into mesophyl protoplasts of Nicotiana tabacum and into recipient cells of wild tomato (Lycopersicon peruvianum) with subsequent selection and regeneration of hybrids are considered. Development of alternative methods of partial genome transfer using microprotoplasts produced from plant gametes (microspores) is presented. The advantages of microprotoplast-mediated transfer of chromosomes as an efficient technique of parasexual crossing in comparison with traditional somatic hybridization are discussed. PMID- 12774517 TI - [Molecular-biological nature of morphological abnormalities induced by chronic irradiation in coniferous plants from the Chernobyl nuclear power plant exclusion zone: emphasis on a possible role of the cytoskeleton]. AB - Results of the analysis of morphological abnormalities in coniferous plants from the Chernobyl exclusion zone are described. It is shown that such features as total and individual protein content, genome organization and peculiarities of its expression, and karyotype are different for the control and morphologically abnormal needles. A possible role of the cytoskeleton structures in the abnormal morphogenesis in plants is discussed. PMID- 12774518 TI - [Role of cytoskeleton in gravisensitivity of a plant cell: experimental data and hypotheses]. AB - The article highlights the hypotheses of plant gravisensing, including those where cytoskeleton is involved. Data concerning arrangement of microfilaments and microtubules in specified and non-specified to graviperception cells of higher and low plants and concepts of the role of cytoskeleton in plant gravisensing are considered. PMID- 12774519 TI - [Comparative analysis of primary structure of the mutant tubulins with resistance to antimicrotubular drugs for prediction of new mutations with analogous properties]. AB - All known for today complete amino acid sequences of alpha- and beta-tubulins were aligned and analyzed to reveal the regularity of location of mutations result in the resistance to antimicrotubular compounds and a prediction of positions of new similar mutations. It was shown, that known positions of amino acid changes lead to decrease a affinity to antimicrotubular agents with depolymerizing mechanism of action are consensus and located in proximity to the residues involved in intradimeric/interdimeric interactions and interactions with nucleotides (within of six residues), but never coincide with them. For changes lead to resistance to stabilizing antimicrotubular compounds, similar dependence is not traced. The identified regularity enables to predict the positions of new mutations lead to resistance to agents with depolymerizing mechanism of action. PMID- 12774520 TI - [Standardization of cancer treatment and legal medical standards]. AB - Legal medical standards are usually applied in two ways in Japanese medical malpractice cases. One application is a judgment on the quality of medical treatment, and the other is an evaluation of the validity of informed consent. Legal medical standards are not equivalent in all medical institutions. If physicians in a medical institution are expected to be aware of the most recent medical knowledge, that becomes the medical standard of the institution. For physicians in major metropolitan medical institutions, such knowledge constitutes the medical standard even before it is published in the guidelines of a committee of an established national medicoprofessional society. The standardization of cancer treatment may be used as evidence in medical malpractice suits brought by cancer patients based on both the quality of treatment and valid informed consent. In addition, it is possible that the validity of informed consent may be given more weight by the courts than the following of a course of standardized treatment by individual physicians/medical institutions in cases where patients have done extensive research on and exhibit informed knowledge of new treatment modalities. PMID- 12774521 TI - [Standardization of lung cancer surgery]. AB - A lobectomy with systematic nodal dissection is accepted as the standard radical operation for early-stage lung cancer. When this was established as the standard operation, there was little evidence to confirm that the results after a lobectomy with systematic nodal dissection were superior to those after other procedures. Most thoracic surgeons accepted lobectomy with systematic nodal dissection as the gold standard for lung cancer surgery. Therefore, no randomized, controlled study was conducted to confirm the justification for that procedure. However, numerous reports of nonrandomized trials or practices were published to justify its acceptance as the standard radical operation for stage IA lung cancers. On the other hand, advances in diagnostic equipment including helical computed tomography (CT), magnetic resonance imaging, and position emission tomography rapidly changed the concept of early-stage lung cancers. Detection of ground glass attenuation by CT and the introduction of the new pathological concept of localized bronchioloalveolar cell carcinoma as an extremely early-stage lung cancer encouraged surgeons to start clinical trials to obtain evidence. Those efforts will create a new standard for lung cancer surgery. Finally, patients will choose a procedure after consideration of his or her own personal situation excluding medical condition. The procedure selected by most patients will become the gold standard. The gold standard will change with changes in society. PMID- 12774522 TI - [Future perspectives on the standardization of surgical treatment for esophageal cancer]. AB - The Japanese Society for Esophageal Diseases published guidelines for the treatment of esophageal cancer in December 2002. Radical surgery is indicated for T1N1 and T2,3 disease without M1 metastasis to other organs, which consists of transthoracic esophagectomy through the right chest with mediastinal and abdominal (two-field), and cervical if necessary (three-field) resection, lymphadenectomy, and esophageal reconstruction by pulling up the stomach. The survival benefit of cervical lymphadenectomy remains controversial. A randomized, controlled trial (RCT) comparing two-field and three-field resection is needed to evaluate the efficacy of cervical lymphadenectomy. In the West, especially in the USA, surgeons prefer transhiatal esophagectomy, which is illogical in cancer surgery, rather than transthoracic esophagectomy. A recent Dutch RCT comparing transhiatal and transthoracic esophagectomy reported lower morbidity and a trend toward improved long-term survival in the transhiatal group. Minimally invasive surgery for esophageal cancer is common in clinical practice today. However, there is little evidence showing that less-invasive procedures are superior to radical surgery. Further investigation is needed to determine the efficacy of thoracoscopic esophagectomy and laparoscopic mobilization of the stomach for esophageal replacement. The efficacy of neoadjuvant chemotherapy and chemoradiotherapy also remains controversial. However, the effectiveness of adjuvant chemotherapy after surgery on disease-free survival was confirmed by the Japanese Clinical Oncology Group RCT. PMID- 12774523 TI - [Perspectives on the standardization of surgical treatment for gastric cancer]. AB - To obtain better outcomes of surgical treatment for gastric cancer, standardization of three major aspects, indications for treatment, surgical technique, and evaluation of the results are required. The practice guidelines for gastric cancer treatment postulated by the Japanese Gastric Cancer Association in 2001 provide optimal indications for treatment, but do not comment on the level of surgical technique and method to evaluate the results. Technical training has been performed in the traditional fashion based on apprenticeship in Japan, and there is no standardized evaluation system of the level of technique. Therefore guarantees of the level of surgical technique should be offered by medical societies and/or authorized associations. Surgical education using simulation system is a promising tool for standardization of the level of surgical technique. In the evaluation of the results of gastrectomy, the survival rate has been widely accepted as an objective evaluation. On the other hand, due to recent advances in the surgical treatment of gastric cancer and the increasing number of cases of early gastric cancer, attention has been paid not only to the survival rate but also to the quality of life(QOL) after gastrectomy. However, the general rules for evaluation of QOL have not been established. The standardization of surgical treatment of gastric cancer based on the estimation of these three aspects may offer a better prognosis and better QOL after gastrectomy. PMID- 12774524 TI - [Perspective of standardization of surgical treatment for hepatocellular carcinoma]. AB - Hepatocellular carcinoma (HCC) is one of the most common malignancies in Japan and worldwide. In recent years, surgical treatment for this disease has made great progress, leading to reduced operative morbidity and mortality. The advances have made it feasible to standardize surgical treatment. Moreover, the public deserves quality treatment ensured by surgical expertise. In these circumstances, the author believes that the following are important issues in the standardization of surgical therapy. 1) Surgical indications and evaluation of hepatic functional reserve, for which the Makuuchi criteria appear useful in principle, must be standardized. 2) The surgical method must be determined. Anatomically systematized resection may be reasonable, although nonanatomical resection may be performed when resection of less extent than the Healey segment is performed. 3) A consensus must be reached on pre- and postoperative adjuvant chemotherapy. Interferon administration liver transplantation may be a promising prophylactic therapy against hepatitis C-related HCC recurrence after curative resection. 4) It is necessary to clarify the long-term survival rate of patients with small HCC who undergo radiofrequency ablation in relation to tumor size, associated liver disease, tumor differentiation, tumor vascularity, etc. Appropriate treatment should be based on an analysis of these four factors. In addition, the issues of surgical indications for vascular tumor thrombus, preoperative portal branch embolization, laparascopic hepatectomy, and also liver transplantation should be discussed, along with perioperative management and postoperative follow-up. PMID- 12774525 TI - [Present situation of and prospects for standardization of surgery for biliary tract cancer]. AB - Surgery remains the primary treatment for biliary tract malignancies. A standard therapy is required in the era of evidence-based medicine. However, guidelines for biliary tract cancer surgery have not yet been adopted. We review the current literature on the treatment of biliary tract malignancies and surveyed the prospects for standardization of biliary tract cancer surgery. For patients with gallbladder cancer, surgical procedures should be selected based on the depth of invasion. In aggressive surgery for stage IV gallbladder cancer, attention should be paid to the high incidence of operative mortality. Hepatopancreatoduodenectomy and extended lymph node dissection have been investigated in clinical trials. For hilar cholangiocarcinoma patients, surgical procedures should be selected based on tumor location. For Bismuth types III and IV, hepatectomy combined with caudate lobectomy appears to be the standard surgery. Liver transplantation for hilar cholangiocarcinoma patients is currently being examined in clinical trials. Pancreatoduodenectomy is considered the standard procedure for middle and lower bile duct and ampullary cancer. It is necessary to continue developing adjuvant therapies for patients with negative prognostic factors. The appropriateness of adjuvant therapies such as chemotherapy and radiation therapy should be confirmed in further clinical trials. It is necessary to reach a definite consensus on standard therapy for biliary tract malignancies in the near future, and therefore multicenter, randomized trials to address the issues involved should be initiated in Japan. PMID- 12774526 TI - [Prospects for standardization of surgical procedures for carcinoma of the pancreas]. AB - Since physicians need to guarantee the efficacy of medical therapy for patients, therapies for patients with cancer should be standardized to some extent. Carcinoma of the pancreas has the highest death rate of all cancers, with a resection rate as low as about 25% to 30% and a 5-year survival rate of around 9%. It is very difficult in such a situation to standardize the surgical strategy for carcinoma of the pancreas. Because pancreatic cancer is a general disease, the treatment strategy should include not only complete surgical resection but also local control methods with intraoperative radiation, prevention of liver metastasis, development of effective anti-cancer drugs, etc. Major progress in therapy for pancreatic carcinoma may be expected in the near future by with the cumulative use of effective therapies. Standard resection and extended resection: For carcinoma of the head of the pancreas, pancreaticoduodenectomy with regional lymph node dissection is performed in Japan, as is extended resection with thorough lymph node dissection of the retroperitoneal and paraaortic region. However, so far the prognosis of patients who undergo extended resection is not better than those who undergo standard resection. A randomized controlled trial of the two types of resection is now being conducted and its results are awaited. For carcinoma of the body and tail of the pancreas, distal pancreatectomy and splenectomy with lymph node dissection is performed if hematogenous or massive lymph node metastasis or direct invasion of the large vessels has not occurred. The Appleby procedure is performed in some cases. Reconstruction and complications of surgical procedures of carcinoma of the pancreas: It appears that a decrease in complications and a lower death rate have been achieved due to pancreaticoduodenectomy rather than due to the extent of lymph node dissection. In particular, progress in anastomosis techniques of the pancreas and intestine and in perioperative control has been marked. For prevention of complications, it is important that absorbable synthetic sutures be used in the pancreaticojejunal anastomosis, that the cut end of the pancreas be sutured and covered by the jejunum without dead space, and that the stent tube be inserted into the main pancreatic duct. The pancreaticojejunal anastomosis should be bordered by the greater omentum. This technique will prevent both the spread of the pancreatic juice into the intraabdominal cavity and rupture of the blood vessels, which can cause fatal postoperative bleeding. Sufficient intraabdominal drains should be in place, especially around the pancreaticojejunal anastomosis. Radiochemotherapy: There are no effective anticancer drugs for the treatment of carcinoma of the pancreas. It was reported that low-dose 5-fluorouracil and cisplatin (5-FU and CDDP) and gemcitabine plus either 5-FU, epirubicin, or CDDP has some effect. The efficacy of intraoperative radiotherapy has not been confirmed. It is not apparent whether radiochemotherapy is superior to surgery. Curable pancreatic carcinoma: Intraductal papillary-mucinous tumors of the pancreas (IPMT) take their name from the histological feature of mucin production and correspond to so called mucin-producing tumors of the pancreas. This tumor is classified into two types, the main pancreatic duct type and the branch type. About 90% of the main pancreatic duct type and 20% of the branch type are malignant. The branch type of IPMT resembles a bunch of grasps in imaging procedures. Approximately 60% of cases with the branch type of IPMT can be followed up without surgery. Since the prognosis of IPMT is fairly good and the 5-year survival after surgery is about 70% to 80%, limited resection of the pancreas with organ preservation is under investigation. Mucinous cystic tumors of the pancreasin are characterized by development in the body and tail of the pancreas in middle-aged women, with histological ovarian-type stroma in the wall of the tumor, and round cystic lesions with a fibrous capsule containing multiple cystic components of various sizes, which resembles a Chinese citron upon imaging procedures. Surgery should be performed if such a diagnosis is made. PMID- 12774527 TI - [Actual standards and controversies in colorectal cancer surgery]. AB - In early colorectal cancer, the standard treatment for superficial carcinoma limited to the mucosa is endoscopic polypectomy or local resection. If the carcinoma invades the submucosa, the standard surgical procedure is bowel resection with lymph node dissection. In advanced colon cancer, the ideal extent of bowel resection is defined by removing the blood supply and lymphatics at the level of the origin of the primary feeding arterial vessels. When the primary tumor is equidistant from two feeding vessels, both vessels should be excised at the origin. It is desirable to remove is more than 10 cm of the bowel on either side of the primary tumor. The value of no-touch isolation is controversial. Laparoscopic-assisted colectomy should be limited to clinical trials. For patients with advanced rectal cancer, 4-6 cm clearmargins from the attached mesorectum distal to the tumor are desirable. The ideal distal margin length is 3 cm or greater from the transected mucosal edge to the distal edge of the primary tumor. The inferior mesenteric artery should be excised at its origin. Extended lateral lymph node dissection is indicated for patients with lower rectal cancer invading the muscularis propria or deeper. In stage IV and recurrent cancer, surgical resection is recommended if it appears to offer cure. PMID- 12774528 TI - [Prospects for standardization of surgical treatment for breast cancer]. AB - Surgical treatment of breast cancer in general guided cancer surgery in the direction of extensive radical operations until two decades ago. More recently, the trend has been toward conservative, less-extensive surgery. Moreover, breast cancer has become a model case for establishing standardization of surgical care of cancer. The Japanese Breast Cancer Society issued "Guidelines for Breast Conserving Therapy" in 1999, and the Clinical Research Group for Establishing Guidelines for Standardized Diagnosis and Treatment for Breast Cancer became active in 2002 under the auspices of the Ministry of Health, Welfare and Labor. There are several areas of controversy in terms of surgical care in breast cancer. Examples are yet-immature breast-conserving surgery which is performed with wide variations (15-90%) depending upon the institution, and sentinel lymph node biopsy which must await long-term results before being recognized as standard care. The installation of expensive diagnostic devices indispensable for standardized surgical care might not be necessary at each institution, and it must be noted that postoperative follow-up policies practiced in Japan differ greatly from those recommended by the American Society of Clinical Oncology. Despite the anticipated difficulties, a consensus on what constitutes standardized surgical treatment for breast cancer might be easily reached among breast cancer surgeons. However, it may be somewhat difficult to popularize the standardized surgical treatment among ordinary general surgeons in Japan, since there is a wide disparity in the level of clinical care offered by individual physicians and healthcare institutions. Standardization of medical care guided by the government is mainly designed to reduce healthcare costs, which is instrumental in eliminating unnecessary medical interventions and may in turn shed light on valuable medical care advances. This may be the moment for the Japanese Surgical Society and its members who practice life-saving surgery to lead a genuine national medical reformation. PMID- 12774529 TI - [When we have a medical accident, what should I do?]. PMID- 12774530 TI - Denial of AOD use: an issue for social workers and the profession. AB - The author investigated AOD use among social workers, examined its potential negative professional consequences and explored the denial that may facilitate AOD abuse. Results are reported from an anonymous survey of 751 NASW members in North Carolina. Twelve percent of the sample were at serious risk of AOD abuse, 28 percent reported binge drinking during the preceding year, and 21 percent had used drugs illegally since becoming a social worker. In addition, 34 percent of respondents at serious risk of AOD abuse reported at least one impairment incident, and 39 percent agreed that they had worked when too distressed to be effective. Only 9 percent of those at serious risk reported current problems with alcohol or other drugs; 28 percent of them reported being in recovery, notwithstanding their current drinking and using. Research and practice implications for social workers, supervisors, and the profession are presented. PMID- 12774531 TI - Readiness to adopt best practices among adolescents' AOD treatment providers. AB - Social work, along with other human services professions, is increasingly moving to establish guidelines for treatment interventions that are based on empirical knowledge about populations, needs, and treatment effectiveness. This work, however, is in the beginning stages. This article explores how substance abuse treatment providers for adolescents attempt to obtain and use information to guide a "best practices" approach to treatment. Focus group data were analyzed for themes indicating supportive attitudes toward adopting and evidence of capacity to implement an evidence-based approach. Although support was fairly strong, capacity was relatively weak. PMID- 12774532 TI - Demographic and clinical correlates of client motivation among substance abusers. AB - Understanding the role of motivation in substance abusers' acceptance of treatment is critically important to improving treatment outcomes. To examine this phenomenon a cohort of substance abuse treatment clients was recruited from two treatment programs whose services are funded by a state managed care system. Motivation at treatment entry was examined. Better motivation was consistently associated with severity of substance use. Neither the coercion that accompanies legal system involvement nor self-referral was significantly related to measured levels of motivation. Motivation was not related to alcohol and drug use severity six months later. The severity associated with motivation at treatment entry was, for the most part, not related to clients' success six months later. Implications for social work practice with substance abusers are discussed. PMID- 12774533 TI - Harm reduction: a social work practice model and social justice agenda. AB - Efforts in the United States to eradicate drug use through supply reduction (that is, the War on Drugs) have increasingly violated the principles of social justice and human rights, both locally and globally. This has created ethical conflicts for social workers in policy making, practice, and research. Harm reduction has been conceptualized as a peace movement and is aligned with the humanistic values around which social work is organized. The authors examine how social workers may reduce the ethical conflicts associated with efforts to address substance abuse by adopting a harm reduction approach to policy, practice, and research. They examine current drug policies, the consequences of the policies, and, in particular, how the policies affect social workers as practitioners, agents of social control, and guardians of social justice. PMID- 12774535 TI - Twelve-step recovery and community service. AB - Twelve-step fellowships have been viewed as possibly inhibiting participants' community involvement. This article reports on a qualitative study of 33 recovering people who attended a 12-step fellowship and involved themselves in the community to find out how this involvement was affected by their 12-step participation. This article describes the participants' community activity and their perceptions of how it related to their recovery process. Respondents reported that the fellowship prepared them for community activity by teaching them skills, giving them opportunities to practice participation with others, motivating them to contribute, and directing them toward community service. Implications for practice are discussed. PMID- 12774534 TI - Oregon's transition to a managed care model for Medicaid-funded substance abuse treatment: steamrolling the glass menagerie. AB - The approval of a Health Care Financing Administration (now called Centers for Medicare and Medicaid Services) 1115 Medicaid waiver in Oregon allowed the state to design and implement an expanded publicly funded health care system, the Oregon Health Plan (OHP). Integral to OHP is the administration of physical and behavioral health services, including outpatient substance abuse treatment, through contracted managed care organizations. The two overarching changes to the outpatient substance abuse treatment system were expanded Medicaid eligibility and new operating procedures for the outpatient substance abuse treatment system. The authors used grounded theory to examine the effects of this transition on the treatment system, with an emphasis on the experiences of treatment providers. PMID- 12774536 TI - Group interventions with low-income African American women recovering from chemical dependency. AB - This article presents findings from an investigation of two group therapy modalities involving 93 women who were generally single, with dependent children and limited education and income levels. The article presents an overview of intervention activities that participants found beneficial. Although the group interventions warrant additional rigorous evaluations, these programs helped participants develop a sense of community, reduce stress, identify and minimize gaps between current circumstances and potential positive outcomes, and improve self-efficacy and the ability to think positively. PMID- 12774537 TI - Psychiatric disorders and substance dependence among unmarried low-income mothers. AB - The study reported in this article examined the prevalence of mental health disorders and the sociodemographic factors associated with having a mental health disorder in a probability sample of 185 African American and white single mothers. Logistic regression analyses revealed that race (being white) and being on welfare were associated with increased risk of having a mental health disorder, when controlling for other sociodemographic variables. The association of welfare status and psychiatric disorders highlights the need for access to mental health services for this population. Implications for low-income women making the transition from welfare to employment are discussed. PMID- 12774538 TI - Project assist: a modified therapeutic community for homeless women living with HIV/AIDS and chemical dependency. AB - This article described a substance abuse treatment program for homeless women who abuse substances and are living with HIV/AIDS. A therapeutic community model was modified to meet the unique needs of this population. However, in light of criticism that the traditional TC model fails to attend to women's issues (Brown et al., 1996), the program uses a gender-specific treatment approach. In addition, the treatment program incorporates a variety of HIV support and education services, and provides enhanced health services to address the multiple medical needs of this population. To date, limited treatment options have been available to address the unique issues of women who are homeless, chemically dependent, and HIV-positive. To our knowledge, this is the first description of such a program to appear in the professional literature. PMID- 12774539 TI - Reality management. Real-time data let hospital leaders base decisions on what's happening now, not last year. AB - Managing quality improvement, patient safety, workforce issues and strategic decision-making makes a lot more sense when you base your decisions on what's happening in the present, not the past. Real-time management is proactive and allows potential mistakes to be corrected before they can hurt patients. PMID- 12774540 TI - To be or not to be vaccinated. The smallpox controversy plays out in California hospitals. AB - The debate continues over whether hospitals will vaccinate their clinicians for smallpox. Some hospitals see the vaccination program as part of their mission, while others are taking a wait-and-see attitude. PMID- 12774541 TI - Real men choose nursing. Nursing schools and hospitals target men in their recruitment efforts. AB - In an effort to expand the nursing workforce, nursing organizations, schools and hospital recruiters are targeting men, who now comprise only 5.4 percent of the current workforce. PMID- 12774542 TI - How can boards fulfill their quality oversight role? PMID- 12774543 TI - Ruby Carter: "Gentle giant". PMID- 12774544 TI - Proactive risk reduction: how it works. PMID- 12774545 TI - Walk the PATH to good health. PMID- 12774546 TI - Tularemia in Delaware: forgotten but not gone. PMID- 12774547 TI - The social impact of medicine. Chapter 6. PMID- 12774548 TI - Physicians emeritus group considers "a better way". PMID- 12774549 TI - "A better way". PMID- 12774551 TI - Reports from the AMA interim meeting. December 2002. PMID- 12774550 TI - Medical musings. PMID- 12774552 TI - Treatment of chronic heart failure in a managed care setting. Baseline results from the Achieving Cardiac Excellence Project. AB - BACKGROUND: Effective therapy for chronic heart failure (CHF) is underutilized despite a broad consensus regarding treatment recommendations. METHODS: As a quality improvement project designed to reduce preventable hospitalizations associated with CHF, we examined use of angiotensin converting enzyme inhibitors (ACEI), angiotensin receptor blockers (ARB), and beta-adrenergic receptor blockers (BB) in a population of patients enrolled in a managed care plan. Medicare and commercial enrollees were included. Patients with CHF were identified using claims data (International Classification of Disease 9th Clinical Modification code 428) covering January 1, 1998 through December 31, 1998. Drug utilization data were obtained from the plan's pharmacy benefits database. Data were available for 1220 patients. RESULTS: The mean age (+/- SD) was 71 +/- 12 years, 53% were female, and 84% were Medicare enrollees. Prescriptions for ACEI, ARB and BB were filled by 52%, 9% and 25% of patients, respectively. Prescriptions for diuretics, digitalis preparations, and calcium channel blockers (CCB) were filled by 69%, 34%, and 32%, respectively. Therefore, almost half of patients with CHF were not receiving ACEI therapy, even though it had been proven to reduce morbidity and mortality related to CHF. Furthermore, three-quarters of patients were not receiving BB therapy, a similarly effective therapy. In contrast, CCB and digitalis have not been convincingly shown to reduce mortality in patients with CHF broadly defined. Utilization of CCB and digitalis exceeded that of BB. CONCLUSIONS: Managed care organizations should develop, test, and implement network-level strategies designed to optimize the appropriate utilization of effective drug therapies for patients with CHF. PMID- 12774553 TI - Cervical cancer in North Carolina. Incidence, mortality and risk factors. PMID- 12774554 TI - The rise and decline of managed care. What comes next? PMID- 12774555 TI - The evolution of an HMO. Hands-on experience for North Carolina physicians. PMID- 12774556 TI - The accountability of managed care. The North Carolina Patient's Bill of Rights. PMID- 12774557 TI - Putting managed care regulation in perspective. PMID- 12774558 TI - Employers and the healthcare marketplace. PMID- 12774559 TI - Accessible, affordable healthcare. A continuing evolution. PMID- 12774560 TI - Is Hawthorne bothering pharmaceutical care research? PMID- 12774561 TI - Preparing the pharmacist for the future: PCT to the rescue. PMID- 12774562 TI - Interventions in seamless care. AB - OBJECTIVE: The aim of this study was to investigate whether good collaboration between community pharmacists and hospital pharmacists prevents medication errors. These errors might occur when a patient is discharged from hospital. Both disciplines can complement each other in medication management for the patient. METHOD: The documented interventions of eight teams consisting of a community pharmacist and a hospital pharmacist, were collected and interpreted with a focus on structural problems and also on positive and negative items, that could influence the intensive collaboration between both disciplines. RESULTS: The registered interventions can be grouped into five categories. Most interventions were registered in the category where the major problem is a lack of communication with the patient. Due to collaboration between the hospital and community pharmacist, all identified problems were resolved properly. CONCLUSION: When communication between community and hospital pharmacists is optimised, patients will face fewer problems with their medication when they are discharged from hospital. PMID- 12774563 TI - A longitudinal study of United Kingdom pharmacists' misdemeanours--trials, tribulations and trends. AB - BACKGROUND: Standards of UK pharmacy practice are maintained by the Royal Pharmaceutical Society of Great Britain, which has the power to take a range of sanctions, including removal of the right to practice, against those found guilty of malpractice. This function is currently under review. OBJECTIVE: To conduct a longitudinal study in order to define trends and identify areas where remedial or preventative support could be focused. METHOD: Case analysis of reports of individuals' misdemeanours published in the British Pharmaceutical Journal over a 12-year period (September 1988-October 2000). Professional and personal misdemeanours were considered. MAIN OUTCOME MEASURE: Nature of misdemeanour, conviction or disciplinary proceedings against individual, practising pharmacists in the study period. Reasons offered for committing the misdemeanour and penalties applied. RESULTS: 344 cases, involving a wide range of personal (162) and professional (590) misdemeanours were found. On an annual basis, the maximum incidence of pharmacists found guilty of any misdemeanour was extremely low (< 0.1 of 1% on the pharmaceutical register). The most common professional misdemeanour was failure to keep adequate written records. The most common personal misdemeanour was fraud. The most common reason cited for committing any misdemeanour was financial gain. Numbers in individual offence categories were persistent but low and there were few obvious trends with time. The odds of involvement ratio for male versus female pharmacists was 7.36 (CI: 5.23-10.35) and for ethnic minority versus Caucasian pharmacists was 3.8 (CI: 3.06-4.72). The most stringent penalties (either imprisonment or removal of the right to practice and frequently both) were applied to cases involving personal use or trafficking of drugs subject to abuse. CONCLUSIONS: The current self-regulation of pharmacy practice in the UK involves a wide range of misdemeanours of varying severity; but the incidence of reports of pharmacists found guilty of malpractice was extremely low. The nature of misdemeanours appeared to change little over the period of the study; this study therefore indicates the spectrum of misdemeanours likely to be encountered by a regulating board in the immediate to medium-term future. If regulatory changes such as competence-based practice rights are introduced, the spectrum may change. PMID- 12774564 TI - Hospital pharmacists' reinforcement of guidelines for switching from parenteral to oral antibiotics: a pilot study. AB - OBJECTIVE: The cost of antibiotics in hospitals may be reduced by streamlining, and, particularly, by early switching from the intravenous (i.v.) to the oral route of administration. The aim of the study was to evaluate the feasibility and impact of guidelines for switching, reinforced by pharmacists. METHOD: Patients admitted to internal medicine wards and treated with i.v. antibiotics for various infections were included for six weeks before (group A) and six weeks after (group B) the intervention. Differences in patient characteristics and their outcomes were sought between the two groups. RESULTS: The 26 patients in group B stayed longer in hospital than the 29 in group A (13.3 vs. 9.7 days; P = 0.05). They also tended to need more time to reach the pre-defined criteria for switching (3.6 vs. 2.4 days; P = 0.09). From that point on, they were switched more rapidly to oral antibiotics (1.5 vs. 3.2 days; P = 0.02), which resulted in a trend toward a lower treatment cost until their discharge (44 vs. 92 euros; P = 0.08). No difference was found between the 2 groups for the duration of the i.v. therapy, or the total in-hospital cost of antibiotics. CONCLUSION: Pharmacists may help implement and reinforce guidelines for switching to oral antibiotics. The evaluation of such interventions implies the choice of appropriate outcomes and the awareness of confounding factors. PMID- 12774566 TI - Frequency and trends of interventions of prescriptions in Flemish community pharmacies. AB - OBJECTIVE: To study the role of the community pharmacist in medication management. METHOD: Participating Flemish pharmacists quantified their interventions during two weeks by use of validated diagrams. The clinical interventions were recorded in detail, while the technical interventions were only registered. RESULTS: 124 pharmacists participated. Intervention was needed in 4.1% of the total number of prescriptions (n = 87.647). On average, 20.2 technical and 8.4 clinical interventions over a period of 2 weeks per pharmacy were needed. The main problem was missing or incorrect data on administering the drug (23%). Missing or incorrect advice (37.8%), dose-related problems (26.1%) and interactions or contra-indications (20.2%) were mentioned as important clinical discrepancies. The pharmacists utilized the patient medication records to solve most of the problems. In one out of five cases, the physician was contacted. CONCLUSION: Interventions by community pharmacists in medication management have been recorded in this study. The results should be translated into a process of integrated pharmaceutical care. PMID- 12774565 TI - Pharmaceutical care: pharmacy involvement in prescribing in an acute-care hospital. AB - BACKGROUND: [corrected] Pharmaceutical care implies reaching a consensus with physicians on prescriptions in cases that call for the substitution of one active ingredient for another, a modification in dose, frequency, route of administration, etc., through the unit-dose distribution system. The goal of pharmacist interventions in the hospital should be to achieve a rational use of drugs; to ensure this, a daily review of patient prescriptions by a pharmacist is necessary. Most of the incidence of drug-related morbidity and mortality is predictable and can be avoided, thus reducing the overall cost of health care and the duration of hospitalization while improving the quality of care. The optimum quality of physician or pharmacist care to be achieved would be one that which maximizes benefits and minimizes risks and costs. OBJECTIVE: The goal of this study was to evaluate pharmacist interventions at the Hospital of Barcelona over a six-month period and their clinical and economic repercussions and the degree of compliance. METHOD: The interventions were recorded on a card and classified by type: antibiotic or thromboembolic prophylaxis; substitution of an active principle not included in the hospital's Pharmacotherapeutic Guide; change in dose or route of administration; therapeutic duplication; dose adjustment of aminoglycosides and vancomycin; and inappropriate treatment duration. The economic evaluation considered the average cost of a hospital stay and of the procedures and diagnostics in 1998 and applied data on published probability rates and drug costs. RESULTS: A total of 3,136 interventions were analyzed prospectively during the study period. The interventions represented savings of 129,058.31 euros. Those that contributed most to these savings were recommendations for antibiotic prophylaxis, thromboembolic prophylaxis and pharmacokinetics studies: 49.4, 47 and 5.7% of interventions, respectively, and 79, 3.6 and 15% of total savings, respectively. CONCLUSION: In general, the degree of acceptance of the interventions was high (88.8%), as a result of the growing compliance by physicians with the hospital's established protocols. It can be concluded that pharmacist interventions have been useful to improve patient care and have been important to help educate physicians on the quality of drug therapy. PMID- 12774567 TI - Repeated nitrate prescriptions as a potential marker for angina pectoris. A comparison with medical information from the Rotterdam Study. AB - OBJECTIVE: The objective of this study was to determine whether pharmacy records of nitrate prescriptions could be used as a marker of angina pectoris. METHOD: This study was conducted within the Rotterdam Study, a prospective follow-up study which started in 1991 and included 7983 elderly subjects. During follow-up, 1601 subjects filled a first prescription for a nitrate and later filled at least one other prescription for nitrates according to pharmacy records. After excluding subjects who started using nitrates in 1991 and who had less than one year of medication history, we took a random sample of 78 subjects (10%). We studied discharge and outpatient cardiologist letters and files from general practitioners for additional information on angina pectoris in these subjects, and allocated patients to one of three categories according to the possibility of the initial diagnosis of angina being correct. RESULTS: From the random sample of 78, additional information was available on 75 subjects. Definite angina pectoris was present in 33, probable angina pectoris in 18, and possible angina pectoris in 19 subjects. Five subjects had no angina pectoris. Therefore, 93% had at least a possible diagnosis while 68% had at least a probable diagnosis of angina pectoris. The positive predictive value of 2 nitrate prescriptions of which at least one was for rescue therapy was 94%. CONCLUSIONS: We conclude that the use of more than one nitrate prescription can be used as a marker for angina pectoris. This marker may be useful in epidemiological studies. PMID- 12774568 TI - Pharmaceutical care in Denmark: perceived importance of medicine-related problems and participation in postgraduate training. AB - OBJECTIVE: To investigate the readiness of Danish community pharmacists to provide pharmaceutical care. Special focus was on information provided to patients on medicine-related problems and participation in postgraduate training. METHOD: National cross-sectional survey distributed to all Danish community pharmacies (N = 288). Response rate 75.7%; non-responders were interviewed by telephone to establish their reasons for not participating. MAIN OUTCOME MEASURES: Prioritization of information to patients regarding medicine-related problems. Postgraduate courses selected by pharmacists during the past three years. RESULTS: In accordance with the profile of Danish community pharmacists, respondents were primarily female and half were below 45 years of age. The main reason given for not responding was lack of time. The prioritization of information to patients regarding medicine-related problems resulted in three clusters of answers. The overall topics of the clusters were: information related to the technical-practical use of medicines; information related to pharmacotherapeutic aspects of medicine use; and information related to the function of the medicine. The clusters accounted for 61.9% of the total variance. The most frequently attended postgraduate courses were on quality assurance and specific diseases. CONCLUSION: The patient information prioritized by Danish community pharmacists is primarily of a technical-practical nature. The postgraduate training pursued is primarily technical without much focus on the philosophy of pharmaceutical care. These factors contribute to the lack of proper readiness of pharmacists to practice pharmaceutical care. PMID- 12774569 TI - A dangerous place to live: from the fog of war to the slime of technology. AB - This article addresses the need for constant vigilance against clever and adaptive enemies, specifically the battle against infections. Medical devices are susceptible to infection and risk management needs to be considered in the context of the underlying science and common sense. PMID- 12774570 TI - Current status of molecular biosensors. AB - Innovations in biosensors have come about as a result of improvements in the biological components and the implementation of microsystems technologies. This article reports on some of the latest product developments in this area and highlights future trends. PMID- 12774571 TI - Digital CMOS sensor chips for media-isolated liquid flow sensing. AB - New digital sensor technology that comes from semiconductor engineering enables media-isolated and extremely precise measurement of small mass flows in medical devices. The capabilities and potential of this technology are described. PMID- 12774572 TI - IVD authorized representatives. AB - Under the In Vitro Diagnostic (IVD) Directive, a manufacturer of IVD medical devices located outside the European Economic Area must designate an Authorized Representative before placing those products on the European market. This article discusses the responsibilities of Authorized Representatives under the IVD Directive and some of the important differences of the role as it is defined in the Medical Device Directive. PMID- 12774573 TI - Some "dos and don'ts" for aspiring standardisers. AB - The first of a series of articles on the important practicalities of standardisation. This month, some "dos and don'ts" when considering writing a standard. PMID- 12774574 TI - Innovation and patents: in vitro diagnostics. AB - Changes in interpretation of patent law mean that in vitro diagnostics companies need to be more alert when making patent applications. PMID- 12774575 TI - User reporting of medical device related incidents. AB - In many countries, health-care professionals are required to notify relevant authorities about certain incidents that can be linked to properties, performance and usability issues of medical devices. This study examines whether the user reporting scheme could add to the information that is already available from manufacturer incident reports. PMID- 12774576 TI - The impact of surface mount FeedThru varistors on medical circuit design. AB - MLVs are an effective tool for designers to meet next generation EMC requirements for medical devices. FeedThru MLVs give designers a new option for highly efficient ESD suppression because of their improved turn-on time characteristics. In addition, FeedThru MLVs give designers highly selective EMI filters to suppress unwanted frequency emissions or isolate the circuit from the frequency spectrum where susceptibility would be a concern. PMID- 12774577 TI - Improving needle-point sharpness in prefillable syringes. AB - Improvement of needle-point sharpness and improved injection comfort for patients remain active areas of research. This article reports on a five-bevel needle for subcutaneous administration with a prefillable syringe, and the results of a clinical trial to verify and validate the bench measure of sharpness. PMID- 12774578 TI - Medical technologies at the dawn of the German health-care reform. AB - How health care should be reformed to address the current economic pressures and deliver a better future. PMID- 12774579 TI - Thermoplastic elastomers in medical applications. AB - TPEs provide medical designers with a broad spectrum of soft-feel, hygienic materials that can readily fulfill accepted medical industry standards with the exception of body implants. Ultimate performance is a combination of tailor-made formulations coupled with innovative design that captures the capability of the material's properties and thermoplastic processing techniques, including combination with other polymers by coinjection moulding or coextrusion. PMID- 12774580 TI - 2D matrix codes and machine vision ensure traceability. PMID- 12774581 TI - Rough justice? PMID- 12774583 TI - Greet and treat: 12 months on. PMID- 12774582 TI - Editorial board's eye view. PMID- 12774584 TI - The faculty in action. PMID- 12774585 TI - Bottling up problems. PMID- 12774586 TI - Ambulance nursing. Part Three. PMID- 12774587 TI - Coming home from war: a literature review. PMID- 12774588 TI - Rescue paramedic style: a training scenario. PMID- 12774589 TI - Poisoning: sodium chloride and sodium bicarbonate. PMID- 12774590 TI - Consultant appraisal in Scotland: where are we now? PMID- 12774591 TI - The prevalence of obesity and undernutrition in Scottish children: growth monitoring within the Child Health Surveillance Programme. AB - OBJECTIVE: To assess whether anthropometric data, routinely collected as part of the Scottish Child Health Surveillance System (CHSP-PS, pre-school children; CHSP S, school age children) could provide a means of monitoring/surveillance for obesity and undernutrition at national and health board level. DESIGN: A survey of 15 health boards and both surveillance systems to identify the nature of data collected, format of data, and extent to which data were accessible (e.g., via Information and Statistics Division of the Common Services Agency). Measurements of weight and height collected as part of the CHSP-PS and CHSP-S were extracted from ISD. They were then audited and missing values or implausible values quantified, and degree of dispersion of values used as an index of quality of measurements. SETTING: Health Board Child Health Surveillance Systems and Information and Statistics Division, Edinburgh. RESULTS: Data on height and weight are currently available for 9 health boards for pre-school children and 4 health boards for school age children. This represents coverage of around 80% of the pre-school child population. Analysis of a data extract from the 39-42 month check in 1998/99, used as an example, revealed that 8% of weight and height data were missing, and approximately 1% were implausible measures. Population and health board level estimates of prevalence of obesity and undernutrition were possible and are presented. Data on height and weight are routinely collected in school age children in all health boards, however only four health boards have growth data electronically available via the school CHSP. CONCLUSIONS: Growth data routinely collected as part of child health surveillance for Scotland can be used to estimate population prevalence of undernutrition and obesity. These can in turn be used to monitor trends at local and national level, to monitor achievement in relation to public health targets, identify risk factors and high risk groups, and to follow cohorts over time. We describe a system of surveillance for undernutrition and obesity and identify its strengths and weaknesses. PMID- 12774592 TI - Glasgow's community care programme: 10 year follow up of discharged patients with schizophrenia. AB - In 1986, the rehabilitation of every long stay psychiatric patient in Glasgow was assessed with a view to reducing bed numbers and developing comprehensive community services. Ten years on, we have attempted to trace 91 patients with a diagnosis of schizophrenia assessed at Gartnavel Hospital, in order to repeat assessments of their psychopathology and levels of functioning. We believe this population represents a unique group in terms of their age, length of hospital stay and chronicity of symptoms. Only two patients were untraced but 36% of the original 91 patients were decreased. Discharge to the community and variations in standards of care appeared to have little impact on the symptomatic presentation of the survivors over ten years. The results provide useful information on the success or otherwise of a large scale discharge and community care programme which is continuing in Scotland. PMID- 12774593 TI - Cardiac troponin T in female athletes during a two-day mountain marathon. AB - BACKGROUND: Equivocal studies exist on the potential of cardiac damage following prolonged endurance exercise. AIMS: The aim of the study was to examine humoral markers of cardiac damage in female athletes during a 2-day mountain endurance race. METHODS: Venous blood samples were drawn from seven female competitors prior to, and immediately following day-1 and day-2 of the event. The serum was analysed for total creatine kinase (CK), creatine kinase isoenzyme MB (CKMB), and cardiac troponin T (cTnT). RESULTS: Elevations in CK and CKMB were apparent following day-1 of the event (mean +/- SD; CK 84.1 +/- 54.6 mg/L vs. 387 +/- 276.7 mg/L, CKMB 2 +/- 1.7 mg/L vs. 5.9 +/- 1.7 mg/L) and subsequently rose further following race completion (CK 743 +/- 500 mg/L, CKMB 11.9 +/- 4.9 mg/L). Elevations in cTnT were noted in three competitors following day-1 cTNT (range 0.013-0.044 mg/L) and remained elevated in two competitors following day-2 (range 0.014-0.017 mg/L). CONCLUSIONS: The elevations in cTnT likely represent release from the cytosolic fraction. The mechanism responsible for such release is yet to be elucidated. PMID- 12774594 TI - Endoscopic surveillance practice for Barrett' s oesophagus in Scotland and early experience in implementing local guidelines. AB - Management of columnar lined oesophagus (CLO; Barrett s oesophagus) is controversial. We prospectively audited surveillance practices in Scotland and prospectively assessed the impact of introducing local guidelines for Barrett s surveillance in Edinburgh. Most respondents were gastroenterologists. The majority take random, not four quadrant, biopsies from the CLO. In Edinburgh during 2000, 80 patients underwent surveillance. The guideline protocol was not followed in 30 (37.5%) patients. Follow up of patients without dysplasia generally conformed to the guidelines. Follow up of patients with low grade dysplasia was highly variable while management of those with high grade dysplasia followed the guidelines. Overall we found a wide variability in the management and surveillance of CLO. Early experience suggests that implementation of guidelines is helpful but there is still variation in practice. PMID- 12774595 TI - Exercise to exhaustion in the second-wind phase of exercise in a case of McArdle's disease with and without creatine supplementation. AB - The cardio-pulmonary and biochemical changes observed in a case of McArdle's disease, exercising with increasing work rates to exhaustion in the "second-wind" phase of exercise are reported for the first time. A work rate of 275-325 watts was achieved. Venous blood lactate remained unchanged throughout. The plasma ammonium level reached a plateau of approximately 400 mmol/l at 100 watts. At a work rate of 150-175 watts the ratio of O2 consumption to CO2 production increased, the inverse of an anaerobic threshold. Maximal cardiopulmonary responses were achieved at 200 watts. During the final periods of exercise from 200 to 275/325 watts pulmonary ventilation did not significantly change but there was a decrease in the venous blood H+ concentration, and pO2 and in increase in the pCO2. Creatine supplementation at 25 g/day for five days did not improve exercise performance. PMID- 12774596 TI - A comparison of travel related ID admissions in Glasgow: (1985; 1998/99). AB - A comparative study was made of patients admitted over 12 months to the principal infectious diseases unit for Glasgow in 1985 and in 1998/99. During this interval UK travel statistics show a 135% growth in visits abroad and a 5% rise to 17% in destinations with a risk of malaria. Travel associated admissions rose by 96% to a total of 108. Patients of Asian/Oriental ethnicity declined from 55% to 18%, whilst Caucasians increased from 38% to 81%. Travellers aged 20-39 years formed the modal age groups (51% and 50%). Gastro-intestinal problems accounted for the largest single diagnostic category in both study periods, 38% and 40% respectively. In-patients diagnosed with malaria fell by 20%; these figures are cause for encouragement to those involved in the teaching and dissemination of advice on malaria prophylaxis. The findings re-enforce the need for the continuation and expansion of travel health education for both healthcare professionals and the public. PMID- 12774597 TI - Isolated hydatid cyst of the breast. AB - Although common in sheep-breeding communities, hydatid disease is actually found worldwide. Man is an occasional intermediate host in whom the disease is manifested by the presence of one or more hydatid cysts, usually located in the liver or lung and rarely at other sites such as the breast, as in our patient. A case of an isolated breast involvement that was diagnosed during surgery is presented and is followed by a brief discussion on the topic. PMID- 12774598 TI - Robert Halliday Gunning and the Victoria Jubilee prizes. AB - More than a century after the death of Robert Halliday Gunning, a large number of lectureships and prizes bearing his name continue to be awarded by scientific bodies and learned institutions in Scotland. Most of these awards were endowed in HM Queen Victoria s Jubilee year (1887-88) and bear the additional qualification 'Victoria Jubilee'. An account of the life of Robert Gunning and his various endowments is complemented by an analysis of the factors which determined the nature of his benefactions. PMID- 12774599 TI - Change of heart. PMID- 12774600 TI - The HSJ interview: Hamilton Anstead. Taking care. Interview by Lyn Whitfield. PMID- 12774601 TI - Delayed discharge. Home truths. AB - The Department of Health wants to turn home improvement agencies into key players in provision of services to older people discharged from hospital. 104 clients have been referred to the project: 55 related to hospital discharge and 49 to admission prevention. Having a single point of contact for a range of home improvement services has made referrals easier. PMID- 12774602 TI - Mental health. Out of bounds. AB - Despite sometimes dramatic results, crisis resolution and home treatment services still face resistance from some mental health teams. Key concerns, which the authors dispute, include applicability to deprived areas, pressure on crisis teams, patient risk and changes to the work of consultant psychiatrists. Co operation across the whole care system is essential to success. PMID- 12774603 TI - When London's found wanting. PMID- 12774604 TI - More premature baby deaths could be prevented. PMID- 12774605 TI - Healthy start could stigmatize low-income mothers. PMID- 12774606 TI - The invisible woman: maternity care in Ireland (1). PMID- 12774607 TI - Birth centres in Wiltshire (1). PMID- 12774608 TI - In Parliament. PMID- 12774609 TI - The midwife and postnatal contraceptive advice (2). PMID- 12774610 TI - Antenatal care--preparing for the birth. PMID- 12774611 TI - Code of professional conduct. Confidentiality. PMID- 12774612 TI - When breast just isn't best. PMID- 12774613 TI - When breast just isn't best. PMID- 12774614 TI - When breast just isn't best. PMID- 12774615 TI - The big pregnancy brain mush myth. PMID- 12774616 TI - The right start? PMID- 12774617 TI - A focus of care for neonatal nursing: the relationship between neonatal nursing practice and outcomes. Part 1. AB - The developmental outcomes of survivors of neonatal intensive care are, in part, a measurement of the quality of care provision. The relationship between neonatal nursing practice and long-term outcomes of survivors remains unclear. This study aimed to determine the focus of care currently influencing qualified neonatal nurses' practice, to ascertain if nursing care is provided in response to the developmental needs and consequent future quality of life of high risk babies. Following a detailed literature review, a case study combining semi-structured interviews with documentary evidence was undertaken within a regional neonatal intensive care unit. The primary focus for the study was the perspective of qualified neonatal nurses working in the field. This first part of a two-part report, presents the background, aims, literature review and design of the study. The findings, discussion and conclusion will be presented in the June issue. PMID- 12774618 TI - 'Getting the right start': the children's national service framework. PMID- 12774619 TI - Developing elective orthopaedic surgery in Malawi. PMID- 12774620 TI - From backwater to beacon: a model for outpatient nursing. PMID- 12774621 TI - Introducing a clinical holding policy. PMID- 12774622 TI - Agenda for change: issues and implications. PMID- 12774623 TI - Applying for ethics committee approval. PMID- 12774624 TI - Promoting health: challenges for children's nurses. PMID- 12774625 TI - Safe administration of medicines to children: Part 1. PMID- 12774626 TI - [Chemical and pharmacological advances of the study on zushima]. AB - OBJECTIVE: To review the progress in the research of the active ingredients of Zushima and their pharmacological activities. METHOD: Base on the articles of the chemical constituents and pharmacological activities of Zushima. RESULT: Traditional Chinese drug, Zushima contains coumarins, diteropenoids, lignans, flavonoids, anthraquinones and sterols. Pharmacological investigation concludes that it has actions of painkilling, antiinflammation, inhibiting bacteria, antithrombus, antitumer and antifertility. CONCLUSION: Zushima has extensive actions in pharmacology. And plant resources are very rich. It is a meaning job to study the chemical ingredients and pharmacological activities of Zushima further. PMID- 12774627 TI - [Molecular authentication of Dendrobium chrysanthum from its allied species of Dendrobium]. AB - OBJECTIVE: To define molecular characters to distinguish D. chrysanthum from its allied species D. primulinum, D. lituiflorum, D. aphyllum, D. crepidatum. METHOD: The molecular characteristics of D. chrysanthum and its allied species were compared. The sequences of rDNA ITS regions were exploited to explore the evidence for authentication D. chrysanthum and its allied species. RESULT: Although the morphological difference was slight, the sequence difference of ITS regions among five rDNAs was obvious and stable. Fifteen sites of ITS region were defined as DNA character to identify D. chrysanthum from the other four allied species. CONCLUSION: The difference of rDNA ITS sequences can be used to authenticate accurately D. chrysanthum from three allied species of Dendrobium. PMID- 12774628 TI - [Comparison among families of Mutong]. AB - OBJECTIVE: To distinguish families of Mutong correctly and direct effective and safe clinical administration. METHOD: Comparison among families of Mutong on Herbs, Taxology, Clinic, Pharmacology and Toxicology. RESULT: 1. There are mainly three families of Mutong: Lardizabalaceae, Ranunculaceae, Aristolochiaceae, which were all included in China Pharmacopeia in 1963. However only Mutong of Ranunculaceae and Aristolochiaceae family have been included in China Pharmacopeia since 1977, but Mutong of Lardizabalaceae family has not been included in China Pharmacopeia ever since. 2. It was Mutong of Lardizabalaceae family that was used mainly through the ages without toxic records, and Mutong of Aristolochiaceae e.g. Caulis Aristolochia manshuriensis (CAM) was not put down in writing of past ages but is mainly used today with toxicity repeatedly. 3. CAM contain aristolochic acid and aristololactam with high toxicity, which plays an uncertain role in diuresis with poor bactericidal power. Mutong of Lardizabalaceae family e.g. Akebia trifoliata (Thunb.) Koidz. var. australis (Diels) Rehd (ATKV) don't contain aristolochic acid and aristololactam, which has low toxicity and plays a certain role in diuresis with high bactericidal power. CONCLUSION: It may be quite safe to use ATKV instead of CAM in clinics. So we suggest that ATKV should be reused as first Mutong in China Pharmacopeia revised edition in order to ensure a correct understanding of the facts and reveal Mutong in its true colors, and CAM should be used as second Mutong strictly according to the rules in China Pharmacopeia revised edition. PMID- 12774629 TI - [Determination of cichoric acid in Echinacea purpuea]. AB - OBJECTIVE: To analyze the natural change rule of active components of E. purpuea by measuring content of cichoric acid. METHOD: Reverse HPLC method was used. RESULT: The maximum cichoric acid content of the roots occured in seedling age of May, and that of the flowers occured in blooming stage of mid July, but cichoric acid in stems was generally low anyway. The maximum content of cichoric acid in the plant above ground occured in the blooming stage of mid July. CONCLUSION: The measuring method of content of cichoric acid is successful and reliable. The optimum stage of harvest in Echinacea purpuea should be guided by natural change rule of cichoric acid content. PMID- 12774630 TI - [Study on the genetiey relationship between Chinese and Korean medicinal materials of niuxi with the method of RAPD]. AB - OBJECTIVE: To study the hereditary relationship between Chinese and Korean medicinal materials of Niuxi achyranthis root. METHOD: Ten samples of four kinds of Niuxi was studied with the method of RAPD. RESULT: Expanded product showed total record of 100 spectrum bands, which proved that the hereditary gap between Korean self-produced A. japonica and A. bidentata is smallest. CONCLUSION: Korean self-produced A. japonica is near to A. Sidentata. PMID- 12774631 TI - [Factors influencing the activity of fibrinolytic enzymes from earthworm, Eeisenia foetida]. AB - OBJECTIVE: To study the factors influencing the activity of fibrinolytic enzymes from earthworm and to obtain the better way to extract fibrinolytic enzymes as well as keep its optimum activity. METHOD: 75% alcohol, 0.9% NaCl and 10% saccharose was used to extract the crude fibrinolytic enzymes from earthworm, the method of urokinase gelose-fibrin plate was used to measure the activity of fibrinolytic enzymes from earthworm. and the method of 3,3'-diaminobezidine tetrahydrochloride colorimetry to was used measure the content of selenium. The method use ts of measuring the content of arsenic was silver diethyldithiocarbamate colorimetry. RESULT: The fibrinolytin of earthworms reared with cattle soils had higher activity than that reared with garbage. The arsenic in the earthworm's body could improve the activity of earthworm's fibrinolytin. However, the selenium had litter influence on it. Among the three methods of extraction, the 75% alcohol one was the most efficient, the 0.9% NaCl was next, and the 10% saccharose was the lowest. The influence of dialysis on the activity of fibrinolytin was less than that of ultrafiltration, when the earthworm's fibrinolytin enzyme was further sublimated. CONCLUSION: The activity of the earthworm's fibrinolytin will be increased earthworm is reared with the fitting baits and when appropriate methods, of extraction and purification are used. PMID- 12774632 TI - [Determination of imperatorin in tetraploidy radix Angelicae dahuricae]. AB - OBJECTIVE: To determine the content of imperatorin in the tetraploidy radix angelicae dahuricae, and compare it with the original diplontic varites. METHOD: The chromatographic method was carried out on Nova-pak (4.6 mm x 200 mm, 5 microns) column with acetonitrile-water solution as a mobile phase. The flow rate was 1 ml.min-1. the detection wavelength was at 248 nm, and the column temperature was 25 degrees C. RESULT: The eontent of was imperatorin in the tetraploidy radix 0.460% and 0.225% imperatorin in the diplontic species. CONCLUSION: The content of the mainly active constituent in the tetraploid is double to what it is in the original diplontic species. PMID- 12774633 TI - [A new steroidal glycoside from fermented leaves of Agave americana]. AB - OBJECTIVE: To investigate the constituents of fermented leaves of Agave americana, and discover new compounds. METHOD: Compounds were purified with silica gel and C8 reverse--phase silica gel column chromatography. The structures were elucidated by chemical and spectroscopic evidence. RESULT: Three steroidal compounds were obtained and their structures were identified as (25R)-5 alpha spirostan-3 beta, 6 alpha, 23 alpha-triol 6-O-beta-D-glucopyranoside(1), (25R)-5 alpha-spirostan-3 beta, 6 alpha, 23 alpha-triol-3, 6-di-O-beta-D-glucopyranoside (cantalasaponin-1) (2) and (25R)-5 alpha-spirostan-3 beta, 6 alpha, 23 alpha triol(hongguanggenin) (3). CONCLUSION: Compound 1 is new compound, named agamenoside C. PMID- 12774635 TI - [Study on pharmacology of ultra-fine particles compound Rehmannia]. AB - OBJECTIVE: To observe pharmacological difference between ultra-fine particles of six ingredient Rehmannia pill and traditional six ingredient Rehmannia pill. METHOD: Pharmacokinetic index was measured by death rate, and pharmacology actions were compared by anti-fatigue, hypoglycemic, clearance rate of charcoal particle, hypoxia resistance and serum hemolysin concentration experiment. RESULT: Dose-effection was significant and pharmacology actions were more than traditional six ingredient Rehmannia pill in six ingredient Rehmannia pill. CONCLUSION: Ultra-fine particles of six ingredient Rehmannia pill are better than traditional six ingredient Rehmannia pill in bioavailability and pharmacology actions, and the weight of pill is reduced while efficacy is enhenced by ultra fine particles. PMID- 12774634 TI - [The studies on the chemical components of trutleback]. AB - OBJECTIVE: Studies on the substance with nourishment Yin for a reasonable and rational auality appraise for turtleback. METHOD: To separate by chromatography and identifying with MS, 1H NMR, 13C NMR, IR. RESULT: Two compounds were separated and identified as hexadecanoyl cholesterol ester and cholesterol. CONCLUSION: The two compounds are isolated from turtleback for the first time. PMID- 12774636 TI - [Studies on anti-endotoxin activity of F022 from Radix Isatidis]. AB - OBJECTIVE: To study the anti-endotoxin activity and mechanism of F022 from Radix Isatidis. METHOD: The production of TNF-alpha and IL-6 of murine peritoneal macrophages stimulated by LPS was measured by ELISA. The temperature in rabbits was tested after i.v. administration of LPS. The lethality of BCG-primed mice was induced by LPS. RESULT: If F022 was added to macrophages culture simultaneously with LPS or 1 h before addition of LPS, production of TNF-alpha and IL-6 by macrophages was remarkably inhibited in vitro. F022 inhibited the fever induced by LPS in rabbits and protected BCG-primed mice from LPS induced lethality if given before administration of LPS. CONCLUSION: The anti-endotoxin effect of F022 may inhibit LPS binding to its receptor, and it may be a LPS receptor antagonist. PMID- 12774637 TI - [Antagonistic effect of aqueous extract of detoxified cottonseeds on corticosterone-induced lesion in cultured PC12 cells]. AB - OBJECTIVE: To study the possible mechanism of aqueous extract of detoxified cottonseeds (CTN-W). METHOD AND RESULT: CTN-W 0.01, 0.03, 0.10, 0.30 mg.mL-1 was incubated directly with the synaptic membrane extracted from the cerebral cortex in rats, and adenylyl cyclase (AC) activity was detected by using radio immunoassay. RESULT: Showed that CTN-W could activate AC in a dose-dependend manner. After incubation with PC12 cells in the presence of corticosterone 2 x 10(-4)mol.L-1 for 48 h, CTN-W 0.08, 0.4, 2 mg.mL-1 protected PC12 cells from the lesion induced by corticosterone. CONCLUSION: Antidepressant and anxiolytic effects of CTN-W are related with the activation of AC-cAMP pathway in signal transduction system, thus protecting neurons from the lesion. These two aspects maybe partly form the mechanism of CTN-W's action. PMID- 12774638 TI - [Study on sober-up effect of crystal sugar-vinegar solution]. AB - OBJECTIVE: To investigate the enhancing effect of crystal sugar-vinegar solution on the tolerance of alcohol consumption in mice and rabbits. METHOD: Crystal sugar-vinegar solution was given to mice or rabbits 30 min before feeding a dose of alcohol. The toxic behavior and percentage of animal death in 24 hours were observed. Meanwhile, blood alcohol levels in the rabbits were measured. RESULT: Crystal sugar-vinegar solution could prolong the latent period of righting reflex disappearing of the drunk mice(P < 0.01) and decrease death percentage of drunk mice in 24 hours(P < 0.01). Crystal sugar-vinegar could also decrease blood alcohol levels in the drunk rabbits, especially 30 min(P < 0.01) and 180 min(P < 0.05) after administration of alcohol. CONCLUSION: Crystal sugar-vinegar solution has an evident sober-up effect on drunk model animal. PMID- 12774639 TI - [Studies on the anti-inflammation effect of the TCM prescription of a combination of monkshood root with peony root]. AB - OBJECTIVE: To make a comparison between the single and combined use of Monkshood Root and Peony Root to observe the anti-inflammation effect in the experimental animals. METHOD: The experimental inflammatory models were adopted, i.e. adjuvant induced polyarthritis carrageenan-induced or formaldehyde-induced rat paw edema, and cotton pellet-induced granuloma formation in rats xylene-induced mouse ear edema, exudation of abdominal blood capillaries of mice, etc. RESULT: The anti inflammafion effect of Monkshood Root was weaker than that of Peony Root or Peony Root combined with Monkshood Root. It was found that anti-inflammation effect with the drug-cooperation was enhanced more significantly in the formaldehyde induced or adjuvant-induceed arthritis models than in the carrageenan-induced rat paw edema and other inflammatory models either in the large dosage of 1:1 proportion or in the small dosage of 1:2 proportion. CONCLUSION: The drug cooperation has a good selective and synergic effect on anti-inflammation. PMID- 12774640 TI - Limiting work hours of residents in teaching hospitals. PMID- 12774642 TI - A model for quality improvement in the neonatal intensive care unit. AB - This article describes a global approach for improving care in the neonatal intensive care unit, using as a case study the process to decrease mortality in extremely low-birth-weight neonates at Lee Memorial Health System, The Children's Hospital of Southwest Florida. Key components to the development of an integrated quality improvement (QI) program--information systems, use of evidence-based medicine, and organizational structure and culture--are presented. The challenge of demonstrating true improvement and the need to better understand the cost of care associated with QI initiatives also are discussed. PMID- 12774641 TI - Differences among hospitals in delivery of care for heart failure. AB - Larger, urban teaching hospitals with larger volumes and greater availability of advanced services provide better care for certain diseases. Because such advanced services have limited importance for routine heart failure management, no hospital type is "disadvantaged." Data on 1,180 congestive heart failure patients were studied to assess the quality of care provided by various types of hospitals. Overall, there was no particular type of hospital that performed consistently better or worse across the quality indicators studied. Substantial opportunities for improvement exist among all hospital types in Tennessee. PMID- 12774643 TI - Interview with a quality leader. Linda Aiken on the healthcare industry and workplace issues. Interview by Susan V. White and Colleen J. Hewes. AB - Linda Aiken, PhD MSN, is director of the Center for Health Outcomes and Policy Research and The Claire M. Fagin Leadership Professor of Nursing and Professor of Sociology at the University of Pennsylvania in Philadelphia. Dr. Aiken's other appointments at the University of Pennsylvania include senior fellow at the Leonard Davis Institute for Health Economics and research associate in the Population Studies Center. Before joining the University of Pennsylvania faculty in 1988, Dr. Aiken served as vice president of the Robert Wood Johnson Foundation. She is a member of the Institute of Medicine, a fellow of the American Academy of Arts and Sciences and of the National Academy of Social Insurance, a former president of the American Academy of Nursing, a distinguished fellow of the Academy for Health Services Research and Health Policy (now AcademyHealth), and a member of the Council on the Economic Impact of Health System Change. Dr. Aiken served on the Medicare Physician Payment Review Commission for 6 years and was a member of the 1982 Social Security Advisory Council and of President Clinton's National Health Reform Task Force. Her recent work, for which she received the Sigma Theta Tau International Clinical Research Award in 1999, has focused on accounting for variation in hospital patient outcomes in the United States, Canada, the United Kingdom, and Germany. Dr. Aiken received her bachelor's and master's degrees in nursing from the University of Florida, Gainesville, and her doctorate in sociology and demography from the University of Texas at Austin. PMID- 12774644 TI - Interview with a quality leader. Sister Mary Jean Ryan on the first Baldrige Award in healthcare. Interview by Susan V. White and Mary Savitsky. AB - Sr. Mary Jean Ryan, Franciscan Sister of Mary, is president/chief executive officer of SSM Health Care (SSMHC), one of the largest Catholic healthcare systems in the United States, with 23,000 employees and 5,000 affiliated physicians serving in 21 hospitals and 3 nursing homes. This year SSMHC became the first healthcare recipient of the Malcolm Baldrige National Quality Award. During her 16-year tenure, Sr. Mary Jean has emphasized three key themes: preservation of the earth's resources, valuing ethnic and gender diversity, and commitment to continuous quality improvement (CQI). She is co-author of CQI and the Renovation of an American Health Care System: A Culture Under Construction, published by ASQ Quality Press in 1997. Sr. Mary Jean has received numerous honors, including the Brotherhood/Sisterhood Award from the National Conference of Community and Justice; (one of) 20 Distinguished Women/St. Louis area; Governor's Quality Leadership Award (Missouri); (one of) 25 Most Influential Women in Business in St. Louis; and the Corporation that Makes a Difference Award from the International Women's Forum. She serves on a number of healthcare and local, state, and national boards. She received a master's degree in hospital and health administration from Xavier University in Cincinnati. She is a nurse and has been a Franciscan Sister of Mary for more than 40 years. PMID- 12774645 TI - Evaluation of a quality improvement intervention for diabetes management. AB - The purpose of this study was to develop and test two interventions designed to improve provider compliance with diabetes management guidelines: the use of a diabetes management flowsheet inserted into patient charts and the use of a diabetes management flowsheet plus quarterly provider feedback about compliance levels. Diabetic patient charts from six family practice clinics were randomly selected and audited at baseline and at 12 months. The analysis indicated that the use of the flowsheet was associated with improved provider compliance in the completion of foot examinations only. Providers involved in the study believed that the process of the flowsheet plus feedback contributed to their greater awareness of diabetes management guidelines. PMID- 12774646 TI - Achieving excellence in veterans healthcare--a balanced scorecard approach. AB - This article provides healthcare administrators and managers with a framework and model for developing a balanced scorecard and demonstrates the remarkable success of this process, which brings focus to leadership decisions about the allocation of resources. This scorecard was developed as a top management tool designed to structure multiple priorities of a large, complex, integrated healthcare system and to establish benchmarks to measure success in achieving targets for performance in identified areas. Significant benefits and positive results were derived from the implementation of the balanced scorecard, based upon benchmarks considered to be critical success factors. The network's chief executive officer and top leadership team set and articulated the network's primary operating principles: quality and efficiency in the provision of comprehensive healthcare and support services. Under the weighted benchmarks of the balanced scorecard, the facilities in the network were mandated to adhere to one non-negotiable tenet: providing care that is second to none. The balanced scorecard approach to leadership continuously ensures that this is the primary goal and focal point for all activity within the network. To that end, systems are always in place to ensure that the network is fully successful on all performance measures relating to quality. PMID- 12774647 TI - Scientific societies and research integrity: what are they doing and how well are they doing it? AB - Scientific societies can play an important role in promoting ethical research practices among their members, and over the past two decades several studies have addressed how societies perform this role. This survey continues this research by examining current efforts by scientific societies to promote research integrity among their members. The data indicate that although many of the societies are working to promote research integrity through ethics codes and activities, they lack rigorous assessment methods to determine the effectiveness of their efforts. PMID- 12774648 TI - Ethics for all: differences across scientific society codes. AB - Ethics codes of a number of scientific societies across different disciplines promulgate ethical standards for responsible conduct in research and other professional activities. The content of these codes of ethics are compared on key dimensions of research, service or practice, and teaching in terms of the range and specificity of the activities these codes cover, and in the degree to which they are educational, aspirational or regulatory in purpose. The role of professional associations in educating, regulating, monitoring, and sanctioning their membership is also discussed. PMID- 12774649 TI - Developing a code of ethics for academics. Commentary on 'Ethics for all: differences across scientific society codes' (Bullock and Panicker). AB - This article discusses the possibilities and pitfalls of constructing a code of ethics for university professors. Professional, educational, legal, and policy questions regarding the goals, format, and content of an academic ethics code are raised and a series of aspirational principles and enforceable standards that might be included in such a document are presented for discussion and debate. PMID- 12774650 TI - Implementing ethics in the professions: examples from environmental epidemiology. AB - The need to integrate ethics into professional life, from the grassroots up, has been recognized, and a comprehensive ethics program has been proposed as a model. The model includes the four dimensions of: consensus building, ethics guidelines development and review, education, and implementation. The activities of the International Society for Environmental Epidemiology (ISEE) are presented as examples and compared with the proposed model. Several innovative activities are described and incentives for ethical professional conduct are highlighted. The examples are provided for emulation by other professional organizations in the hope that, thereby, greater protection of the public interest will be achieved. PMID- 12774652 TI - Establishing an ethical climate in support of research integrity: efforts and activities of the American Sociological Association. AB - The article provides an overview of the recent efforts and activities of the American Sociological Association (ASA) to keep its Code of Ethics visible and relevant to its membership. The development process and challenges associated with the most recent revision of the ASA's code are reviewed, the current education and support activities are described, and other strategies for taking a proactive and leadership role in establishing an ethical climate are proposed. In conclusion, while the ASA has made significant progress in this area, it recognizes that a lot of work remains. PMID- 12774651 TI - Implementing ethics in the professions: preparing guidelines on scientific communication for the Society for Neuroscience. Commentary on 'Implementing ethics in the professions: examples from environmental epidemiology' (Soskolne and Sieswerda). AB - In 1994, the governing council of the Society for Neuroscience was asked to make a brief statement on an issue regarding responsible conduct in publishing. The present article reviews how that initial request grew over the next four years into a lengthy document. Drawing on that experience, which was presided over by the author, comments are made about the potential impact of such guidelines, the lessons learned, and the proper role of professional societies in promoting responsible conduct in research. PMID- 12774653 TI - American College of Epidemiology Ethics Guidelines: foundations and dissemination. AB - Epidemiology is a core science of public health, focusing on research related to the distribution and determinants of both positive and adverse health states and events and on application of knowledge gained to improve public health. The American College of Epidemiology (ACE) is a professional organization devoted to the professional practice of epidemiology. As part of that commitment, and in response to concerns for more explicit attention to core values and duties of epidemiologists in light of emerging issues and increased scrutiny of epidemiology, the College developed, adopted, and published a set of Ethics Guidelines. The structure of the ACE ethics guidelines is in four parts: (1) a brief statement of core values and duties of epidemiologists, coupled with the virtues important to professional practice; (2) concise statements of key duties and obligations; (3) exposition of the duties and obligations with more applications; and (4) a brief summary and conclusion. The Guidelines have been published on the ACE website and in the official College journal Annals of Epidemiology. The guidelines contain (and maintain) core elements that define the discipline of epidemiology and its fundamental duties, but they are also intended to be dynamic and evolving, responsive to a changing professional and social environment. PMID- 12774654 TI - Promoting research integrity at the American Society for Microbiology. AB - The American Society for Microbiology addresses issues of research integrity in several ways. There is a Code of Ethics for Society members and an Ethics Committee, a Publications Board has editorial oversight of ethical issues involved in Society journals and other publications, and the Public and Scientific Affairs Board is involved in ethical issues and scientific policies at the national level. In addition, the Society uses meetings and publications to inform and educate members about research integrity. PMID- 12774655 TI - Roles for scientific societies in promoting integrity in publication ethics. AB - Scientific societies can have a powerful influence on the professional lives of scientists. Using this influence, they have a responsibility to make long-term commitments and investments in promoting integrity in publication, just as in other areas of research ethics. Concepts that can inform the thinking and activities of scientific societies with regard to publication ethics are: the "hidden curriculum" (the message of actions rather than formal statements), a fresh look at the components of acting with integrity, deviancy as a normally occurring phenomenon in human society, and the scientific community as an actual community. A society's first step is to decide what values it will promote, within the framework of present-day standards of good conduct of science and given the society's history and traditions. The society then must create educational programs that serve members across their careers. Scientific societies must take seriously the implications of the problem; set policies and standards for publication ethics for their members; educate about and enforce the standards; bring the issues before the members early and often; and maintain continuing dialogue with editors. PMID- 12774656 TI - Can authorship policies help prevent scientific misconduct? What role for scientific societies? AB - The purpose of this article is to encourage and help inform active discussion of authorship policies among members of scientific societies. The article explains the history and rationale of the influential criteria for authorship developed by the International Committee of Medical Journal Editors, examines questions about those criteria that emerge from authorship policies adopted by several U.S. medical schools, and summarizes the arguments for replacing authorship with the contributor-guarantor model. Finally, it concludes with a plea for scientific societies to play a prominent role in the ongoing debates about authorship and the alternatives as part of their efforts to encourage ethical conduct among their members. Whether or not scientific societies develop authorship policies of their own, they should undertake vigorous educational efforts to keep their new members adequately informed about the importance of authorship practices in ethical scientific research and publication. PMID- 12774657 TI - Challenges in studying the effects of scientific societies on research integrity. AB - Beyond impressionistic observations, little is known about the role and influence of scientific societies on research conduct. Acknowledging that the influence of scientific societies is not easily disentangled from other factors that shape norms and practices, this article addresses how best to study the promotion of research integrity generally as well as the role and impact of scientific societies as part of that process. In setting forth the parameters of a research agenda, the article addresses four issues: (1) how to conceptualize research on scientific societies and research integrity; (2) challenges and complexities in undertaking basic research; (3) strategies for undertaking basic research that is attentive to individual, situational, organizational, and environmental levels of analysis; and (4) the need for evaluation research as integral to programmatic change and to assessment of the impact of activities by scientific societies. PMID- 12774658 TI - The role of scientific associations in promoting research integrity and deterring research misconduct. Commentary on 'Challenges in studying the effects of scientific societies on research integrity' (Levine and Iutcovitch). AB - The nature of scientific societies' relationships with their members limits their ability to promote research integrity. They must therefore leverage their strengths as professional organizations to integrate ethical considerations into their ongoing support of their academic disciplines. This paper suggests five strategies for doing so. PMID- 12774659 TI - Co-responsibility for research integrity. AB - To enlarge the discussion of scientific responsibility for research integrity, this paper offers two historico-philosophical observations. First, in the broad history of ideas, modern ethics replaces social role responsibility with appeals to abstract principles; by contrast, discussions within the scientific community of responsibility for research integrity constitute a rediscovery of the continuing vitality of role responsibility. This is a rediscovery from which philosophy itself may benefit. Second, within the context of scientists' concerns, the idea of role responsibility has undergone significant evolution from "collective responsibility" to the notion of responsibility resting with a "trans-scientific community." Further challenges nevertheless remain in order to relate scientific role responsibility for scientific integrity to the relationship between science and society. To promote a notion of integrity not just in science but in the science-society relationship, it may be useful to think in terms of a "co-responsibility" for scientific integrity. PMID- 12774660 TI - [Hydrolysis of fatty acid esters and glycerol in the blood. Diagnostic value of determining the post-heparin lipoprotein lipase]. PMID- 12774661 TI - [Effect of diquertin on the intensity of lipid peroxidation in patients with diabetes mellitus in the treatment of nail mycosis]. PMID- 12774662 TI - [Content of lipid peroxidation products, alpha-tocopherol and ceruloplasmin in the blood of patients with vascular complications of insulin-dependent diabetes mellitus]. AB - The content of lipid peroxidation products (LPP), as well as the concentrations of alpha-tocopherol (alpha-TC) and of ceruloplasmin (CP) were examined in the blood serum of patients with insulin-dependent diabetes mellitus (IDDM) without any vascular complications and with diabetic micro- and macroangiopathies. It was established that IDDM is accompanied by an accumulation of LPPs and a reduction of alpha-TC in the blood serum. The development of microangiopathies leads to a relative limitation of LPP, still, the intensity of the process exceeds the normal value. If macroangiopathies are available additionally, this aggravates the deficit of endogenous antioxidants due to an extra reduced concentration of the circulating alpha-TC and a simultaneously decreased CP level. PMID- 12774663 TI - [Determination of gamma-aminobutyric acid concentration and activity of glutamate decarboxylase in blood serum of patients with multiple sclerosis]. AB - The pathology of neurotransmitters is of the key importance in the pathogenesis of multiple sclerosis. The developed method makes it possible to determine, with a high accuracy degree, the level of gamma-aminobutyric acid (GABA) and the activity of glutamate decarboxylase (GAD) in the blood serum of patients with multiple sclerosis. The study showed that the level of neurotransmitters and the GAD activity in the blood serum of patients was reduced as compared to the controls. The neuronal transmission pathology can have the key importance in the development, progressing and clinical course of the demyelization process. PMID- 12774664 TI - [Lactoferrin and its role in the pathogenesis of tick-borne encephalitis]. AB - The content of lactoferrin (LF) was studied in the liquor and blood of patients with tick-borne encephalitis (TBE) of the meningeal and focal types; additionally, the information density of the discussed parameter was assessed for evaluating the severity and degree of the inflammation process in the central nervous system (CNS). The LF level was determined in liquor of 37 samples obtained from TBE patients (main group) and of 10 persons with osteochondrosis (controls); it was also determined in the serum taken from 21 TBE patients and from 40 healthy donors by using the immune-enzyme analysis. The LF concentration in TBE patients was found to exceed the normal value by 1.5-3 times during the whole observation period. As for the liquor, it was high, by the onset of the disease, by more than 20 times, however, after the 7th day it was higher 6-fold. A direct dependence of a concentration of the studied protein on a form and severity of the disease was established. The LF level in the liquor of TBE patients alongside with clinical signs can be an objective indicator of a severity and activity of the inflammation process in the CNS; it can also be a criteria of how much the conducted therapy effective is. PMID- 12774665 TI - [Diagnostic strategy of the primary care physician in microhematuria]. AB - The laboratory tests of the urine is of a great importance in the modern clinical practice. The detection and examination of peculiarities as well as of quantitative and qualitative parameters of erythrocyturia plays a significant role in the diagnostic tactic of practitioner. PMID- 12774666 TI - [Current aspects of the pathogenesis and diagnosis of chronic lymphocytic leukemia (review of literature)]. PMID- 12774667 TI - [Errors in determining blood groups (lecture)]. PMID- 12774668 TI - [Soluble transferrin receptors: significance and diagnostic value in anemia]. AB - The study was undertaken for the purpose of demonstrating a diagnostic importance of determining the level of soluble transferrin receptors not only in the diagnosis of iron-deficiency conditions (IDC) or in the differential diagnosis of anemia in a chronic disease (ACD) and of iron-deficiency anemia (IDA) but also in making a diagnosis in case of transfusion-dependent patients with Cooley's anemia. The iron metabolism (including determination of the serum iron (SI), of the total iron-binding ability (TIBA), of saturation of transferrin with iron (STFI), of serum transferrin (ST), and of soluble transferrin receptors (s-STR) was examined in 31 patients with different-genesis anemias, aged 3 to 7. Diagnoses were made for all children on the basis of the standard clinical-and laboratory examinations and tests. A pattern of iron-metabolism parameters, typical of this pathology, was detected in all IDC patients; it is noteworthy, that the s-STR concentration amounted in this category to 9.4 +/- 1.35 mg/l, which essentially topped the normal values. It can be concluded on the basis of the obtained results that s-STR can be regarded as a key IDC marker. There was an increased SF (up to 324 +/- 64.5 mkg/L) alongside with hypoferremia in the ACD patients; the s-STR content was found to be within the reference values (3.21 +/- 0.55 mg/L), the determination of s-STR can be regarded as a test in the differential diagnosis of IDC and ACD. The results of s-STR research ranged, in patients with Cooley's anemia, drastically from 0.8 mg/L to 17 mg/L; it is noteworthy, that there was a reliable dependence on an adequacy of the conducted therapy in case of such patients. The highest sSTR values were found in children with the hereditary spherocytic hemolytic anemia (11.85 +/- 2.5 mg/L), which is, apparently, explained by a proliferative super-activity of the bone marrow observed in this pathology. PMID- 12774670 TI - [Comparative analysis of methods for laboratory diagnosis of Lyme's disease in early stages of infection]. AB - Different methods of laboratory diagnostics were comparatively analyzed in examining 25 patients at the early infection stage. Sera were measured by using various serological reactions. Specific antibodies were determined by using the reaction of indirect immune-fluorescence (RIIF), the immune-enzyme analysis (IEA) and the complement-binding reaction (CBR) in 83.1%, 54.4% and 12.5% of cases, respectively. Essential differences in sensitivity were detected between the above methods. RIIF was proven to be a reliable and sufficiently sensitive method in the laboratory diagnostics of Lyma's disease. While the use of two methods, i.e. RIIF and IEA, ensures the highest percentage of detection of antibodies to the causative agent of Lyma's disease. PMID- 12774669 TI - [Enzymes of purine metabolism of lymphocytes and eosinophiles in bronchial asthma]. AB - The activity of the enzyme purine metabolism (5'-nucleotidase, adenosine deaminase, xanthine oxidase and the content of uric acid, i.e. the final product of the purine metabolism) were determined in lymphocytes, eosinophiles and blood serum of patients with bronchial asthma. The activity of 5'-nucleotidase in lymphocytes, eosinophiles and blood serum of patients with bronchial asthma was found to be reduced by 5-7-% as compared with the controls. The activity of adenosine deaminase in lymphocytes, eosinophiles and blood serum was found to be reduced in a majority of patients. An increased activity of adenosine deaminase was higher in lymphocytes and blood serum of 38% of the examined patients. The activity of xanthine oxidase in lymphocytes and eosinophiles of patients exceeded by 2.4 and 1.7 times the control value, respectively. The content of urine acid was found to be increased, in bronchial asthma, by 2.7 times. Consequently, the balance of enzyme reactions of the purine metabolism is impaired in bronchial asthma, and there is an accumulation of urine acid. PMID- 12774671 TI - [Detection of antigens with the aid of the aggregate-hemagglutination reaction of different serotypes of Ureaplasma urealyticum in the blood serum of patients with inflammatory diseases of the urogenital tract and disorders of reproductive function]. AB - A total of 110 samples of the blood serum, obtained from persons with inflammatory diseases of the urogenital tract and with disorders of the reproductive function were examined by the aggregate-hemagglutination reaction for antigens of the 3d and 8th serotypes, U.u., belonging, respectively, to biovars Parvo and T-960. Antigens U.u. of the 3d, 8th or of both serotypes were detected in 75 (68.18%) of the studies samples. There were no antigens U.u. of the 3d or 8th serotypes in 35 (31.82%) samples. Antigens of biovars Parvo and T 960 were detected at an approximately identical frequency rate. Besides, the mentioned antigens were found in almost one half of blood sera positive to the U.u. samples (48%). No significant differences between the concentrations of antigens Parvo and T-960 were detected in the blood serum samples in case both of them were found in one sample or only one ureaplasma serovar was found in a sample. PMID- 12774673 TI - [Method of making preparations for cytological refractometric examination of glanders]. PMID- 12774672 TI - [Study of herpes viruses in the blood of patients with cardiovascular pathology]. AB - The association between Cytomegalovirus (CMV) and Herpes simplex (HSV-1), on the one hand, and the coronary heart disease and chronic venous insufficiency (CVI), on the other hand, was studied; besides the infection rate by the above viruses in different age groups of donors was examined. The following methods--immune enzyme analysis and detection of viral DNA by means of the polymerase chain reaction (PCR) as well as immune-fluorescence scanning of atherosclerotic plaques -were made use in the study. The pathogenesis of coronary heart disease was shown to be related with the viral contamination caused by CMV, while there was no such correlation between the former and the CVI patients. Finally, it was established that, with age, the frequency rate of seropositive reactions to CMV and HSV-1 was increasing in virtually healthy people. PMID- 12774674 TI - Problem based learning and medical malpractice: does how you've been trained make a difference? AB - Testing the hypothesis that physicians trained in problem based learning formats versus traditional lecture based formats develop equally strong physician-patient relationships, the rates of malpractice filings against graduates trained in each format at the John A. Burns' School of Medicine were compared. With the graduation of 10 more PBL classes, statistically significant differences between the two groups could be obtained. PMID- 12774675 TI - A 76 year-old man with anemia, lymphadenopathy and pericarditis. PMID- 12774677 TI - Childhood cancer. Rapid changes in the last 40 years. PMID- 12774676 TI - The role of violence prevention education at the University of Hawaii John A. Burns School of Medicine (UH-JABSOM). PMID- 12774678 TI - Unsuitable anger. PMID- 12774679 TI - Retention and destruction of medical records. PMID- 12774680 TI - TMA joins the "count on me!" coalition. PMID- 12774681 TI - Medical education: conquering the challenges top concerns and issues in 2003. PMID- 12774682 TI - Tort reform in Tennessee: the next "crisis" state? PMID- 12774683 TI - Declining income--is the solution to hire a consultant? PMID- 12774684 TI - How we are perceived. PMID- 12774685 TI - Bilateral pneumothorax as a complication of changing tracheostomy tube. AB - A 43-year-old man with adult respiratory distress syndrome, secondary to pneumonia, developed bilateral pneumothorax on the fifth post-tracheostomy day, when the tracheostomy tube was being changed to a larger size tube in order to control bleeding from the tracheostomy site. This case demonstrates the potential danger of changing the tube in the postoperative period. PMID- 12774686 TI - The uncivil war against health care fraud. Who will be excluded next? Part. I. PMID- 12774687 TI - Obesity: a growing and serious epidemic for children and adolescents. PMID- 12774688 TI - 2002 survey results of immunization status of 24-month-old children. PMID- 12774689 TI - You and your hearing-impaired patient. PMID- 12774690 TI - Predicting oral absorption and bioavailability. PMID- 12774691 TI - Virtual screening of virtual libraries. AB - Virtual screening of virtual libraries (VSVL) is a rapidly changing area of research. Great efforts are being made to produce better algorithms, selection methods and infrastructure. Yet, the number of successful examples in the literature is not impressive, although the quality of work certainly is high. Why is this? One reason is that these methods tend to be applied at the lead generation stage and therefore there is a large lead-time before successful examples appear in the literature. However, any computational chemist would confirm that these methods are successful and there exists a glut of start-up companies specialising in virtual screening. Moreover, the scientific community would not be focussing so much attention on this area if it were not yielding results. Even so, the paucity of literature data is certainly a hindrance to the development of better methods. The VSVL process is unique within the discovery process, in that it is the only method that can screen the > 10(30) genuinely novel molecules out there. Already, some VSVL methods are evaluating 10(13) compounds, a capacity that high throughput screening can only dream of. There is a huge potential advantage for the company that develops efficient and effective methods, for lead generation, lead hopping and optimization of both potency and ADME properties. To do this, it requires more than the software, it requires confidence to exploit the methodology, to commit synthesis on the basis of it, and to build this approach into the medicinal chemistry strategy. It is a fact that these tools remain quite daunting for the majority of scientists working at the bench. The routine use of these methods is not simply a matter of education and training. Integration of these methods into accessible and robust end user software, without dilution of the science, must be a priority. We have reached a coincidence, where several technologies have the required level of maturity predictive computational chemistry methods, algorithms that manage the combinatorial explosion, high throughput crystallography and ADME measurements and the massive increase in computational horsepower from distributed computing. The author is confident that the synergy of these technologies will bring great benefit to the industry, with more efficient production of higher quality clinical candidates. The future is bright. The future is virtual! PMID- 12774692 TI - gamma-Secretase inhibitors--from molecular probes to new therapeutics? PMID- 12774693 TI - The discovery and development of 5-HT-terminal autoreceptor antagonists. PMID- 12774694 TI - Orally bioavailable beta 3-adrenergic receptor agonists as potential therapeutic agents for obesity and type-II diabetes. PMID- 12774695 TI - Antagonists of the corticotropin releasing factor receptor. PMID- 12774696 TI - Phosphodiesterase type 5 (PDE5) inhibitors. PMID- 12774697 TI - International Nurses Day. PMID- 12774698 TI - Drive to cut hospital infection deaths. PMID- 12774699 TI - The essence of perioperative care. PMID- 12774700 TI - Implementing research in practice. PMID- 12774701 TI - Assistants in surgical practice. What's in a name? PMID- 12774702 TI - Short-term ventilation of critically ill patients in recovery. PMID- 12774703 TI - Breastfeeding the premature infant. PMID- 12774704 TI - A 9-year-old boy with bizarre behavior and growth delay. PMID- 12774705 TI - Maximizing the benefits of human milk feeding for the preterm infant. PMID- 12774706 TI - The role of the pediatrician in extended breastfeeding of the preterm infant. PMID- 12774707 TI - Supporting lactation in mothers with very low birth weight infants. PMID- 12774708 TI - Breastfeeding the borderline (near-term) preterm infant. PMID- 12774709 TI - Medications in breastfeeding mothers of preterm infants. PMID- 12774710 TI - Physiology of lactation in preterm mothers: initiation and maintenance. PMID- 12774711 TI - Reflections on the baby-friendly hospital initiative. PMID- 12774713 TI - Gila Regional Medical Center doubles as art gallery. Open house marks southwestern New Mexico hospital's expansion. AB - Gila Regional Medical Center, Silver City, N.M., is home to a unique kind of art gallery. Though the small town boasts 30 art galleries, one more was added when the newly expanded and renovated hospital opened its doors to the public in February. More than 100 pieces of loaned art estimated to be worth more than $12,000 are on exhibit, in an effort to create a more healing atmosphere for the hospital. PMID- 12774712 TI - St. Mary cooks up awareness with heart-healthy booklet, television. AB - St. Mary Medical Center, Langhorne, Pa., distributed a half-million copies of its copyrighted booklet, "Heart Healthy Living" as the first of a larger, long-term marketing initiative to raise awareness of the suburban medical center. In addition to the medical center and physicians' offices, St. Mary had the booklet distributed by regional food markets and Fleet Bank. These partnerships and those with food products manufacturers helped reduce expenses. St. Mary physicians appeared on a cable television cooking show as well as in selected grocery markets. PMID- 12774714 TI - Opening of Hillman Cancer Center reaps generous media coverage. UPMC News Bureau enjoys results of ongoing efforts. AB - The new Cancer Institute (UPCI), research facility of the UPMC (University of Pittsburgh Medical Center) Health System opened in October 2002. Named The Hillman Cancer Center after noted local philanthropists, it provides a natural opportunity for the UPMC Health System News Bureau to continue its educational goals. Extensive media coverage was valued at nearly $300,000. PMID- 12774715 TI - Parma Hospital raises awareness of new outpatient surgical center. Thanks to continuing coverage, the patients are lining up. AB - Parma Community General Hospital, Parma, Ohio, opened a new ambulatory surgery center Jan. 20 with an awareness-generating campaign. The SurgiCenter was promoted with print advertising in the local newspapers, backed up by the hospital's newsletter, Informed--sent quarterly to 100,000 homes. The response has been beyond expectations. PMID- 12774716 TI - Film shows hospital's neighborliness. Depicts Abbott Northwestern's work with Phillips neighborhood. AB - Abbott Northwestern Hospital, Minneapolis, worked with Twist, a production company also located in Minneapolis, to create a six-minute documentary about its role in the community. It affirms the hospital's commitment to the Phillips neighborhood and uses interviews with local residents, hoping to stimulate community action. PMID- 12774717 TI - Blog your employees, medical staff. AB - Publisher Donald E. L. Johnson shares his insights on blogging, offering ideas that will help communications managers to effectively use this new medium for employees and staff members. PMID- 12774718 TI - The many paths of physician executives. PMID- 12774719 TI - Wandering in the desert. Lessons from a life in health care. AB - A physician executive who's worked in many different areas of health care explains the nearly universal problems he's encountered with the "broken" system. He also suggests ways to fix it. PMID- 12774720 TI - Switching to the dark side?--moving from clinical care to finance. PMID- 12774721 TI - 15 lessons learned: a journey from pediatrics to corporate America. PMID- 12774722 TI - Physician executives as opinion leaders in biotechnology and pharmaceuticals. PMID- 12774723 TI - Pa. malpractice reforms could set the pace for other states. PMID- 12774725 TI - The dead moose and other reflections on compassionate physician leadership. PMID- 12774724 TI - Cardiology practice proves that electronic medical records do raise revenue. PMID- 12774726 TI - Does patenting genes change the meaning of life? PMID- 12774727 TI - Revenue cycle management--Part I. PMID- 12774728 TI - Managing the digital hospital. PMID- 12774729 TI - Being the best you can be. PMID- 12774730 TI - Using simple rules to achieve strategic objectives. PMID- 12774731 TI - Physician executive offers advice from the front lines. Interview by Mary Frances Lyons. PMID- 12774732 TI - Covering the uninsured: the debate returns. PMID- 12774733 TI - Prevalence and care of diabetes mellitus in the Medicare population of North Carolina. Baseline findings from the Medicare Healthcare Quality Improvement Program. AB - OBJECTIVE: To describe diabetes and its care in the Medicare population of North Carolina. STUDY DESIGN: Cross-sectional. DATA SOURCE/STUDY SETTING: Medicare claims for North Carolina residents 18 to 75 years of age were reviewed to characterize diabetes prevalence during the late 1990s and to evaluate adherence to diabetes clinical practice recommendations. PRINCIPAL FINDINGS: Almost 84,000 persons with diabetes and Medicare were identified. Diabetes prevalence was 15.0% overall, 22.9% among African Americans, and 12.9% among Caucasians. A "diabetes belt" of relatively high prevalence was noted in the eastern coastal plain. Glycated hemoglobin testing, lipid testing, and retinopathy screening were performed at about half the recommended rate. Diabetes care rates were lower for African Americans than for Caucasians. CONCLUSIONS: Diabetes is a common and under-treated condition in the Medicare population of North Carolina, especially among African Americans. RELEVANCE: There is much room for improvement in diabetes care within one of North Carolina's most vulnerable populations, the disabled and elderly enrolled in Medicare. Substantial public health opportunities exist in the identification and removal of barriers to diabetes care. PMID- 12774734 TI - Patient-centered outcomes of diabetes self-care. Associations with satisfaction and general health in a community clinic setting. AB - BACKGROUND: Despite the recognition of strategies for diabetes care management, information systems that screen patient diabetes self-care problems are not commonplace, especially among low-income patients. This study examined correlates of three patient-centered outcomes of diabetes self-care (awareness of self-care components, difficulties in diabetes self-care, and adherence to self-care) among low-income diabetic patients. METHODS: Clinical and patient survey data were obtained on 249 participants from 11 primary care community health clinics serving low-income populations in North Carolina in Project IDEAL (Improving Diabetes Education, Access to Care and Living). Measures of patient-centered outcomes of self-care were obtained from the Diabetes Quality Improvement Project (DQIP) Patient-Reported Measures set. RESULTS: A substantial proportion of patients reported difficulty achieving standard self-care treatment goals. The study identified distinct patient characteristics associated with poor self-care outcomes. Increased understanding of self-care components and adherence to them were associated with increased perception of quality of care and, in turn, better general health perceptions in these patients (p < 0.01). CONCLUSIONS: This study identified an unmet need for diabetes self-care knowledge and skills associated with patient outcomes in low-income patients. Routine monitoring of patient centered self-care outcomes could help improve long-term outcomes of diabetes care in this population. PMID- 12774735 TI - Diabetes care in North Carolina. Are we on track for Healthy People 2010? PMID- 12774736 TI - Waging battle against the epidemic. Improving diabetes care in North Carolina. PMID- 12774737 TI - Why are we surprised by our failures? The bewildering complexity of diabetes care. PMID- 12774738 TI - Diabetes self-management training. A nursing perspective. PMID- 12774739 TI - The deadly sins and diabetes. AB - Type 2 diabetes is a spectacularly large and growing health problem. Patients with diabetes have healthcare expenditures that are twice those of the average citizen. We do not clearly understand the genetic predisposition that allows type 2 diabetes to develop, but we do know that lifestyle factors are the most important modifiable factors leading to that disorder. A proper diet and regular exercise (manifested by the presence of a healthy body mass index) can prevent development of latent diabetes, or reverse the manifestations of overt diabetes. It is unlikely that the food industry-already bloated by the high profits from enticing people to buy and consume more and more calories-will voluntarily decide to limit its income. So we will need a number of initiatives from healthcare professionals and from governments at all levels of society to turn this situation around. Some of the measures will require enormous political courage and foresight on the part of our elected officials. Without that, we will hardly need to worry about defense from external enemies. As a nation, we will soon be so fat that we will not be able to waddle to work or to war; those who can will be trying to earn enough money to pay for all the glucose meters and insulin needles and hypoglycemic, antihypertensive, and cholesterol-lowering drugs "required" for the evidence-based treatment of hundreds, even thousands, of blind, neuropathic amputees crowding our cardiac care and dialysis units. PMID- 12774740 TI - Overweight and obesity epidemic in NC. PMID- 12774741 TI - Overweight and obesity epidemic in NC. PMID- 12774742 TI - [The future of surgery, based on scientific evidence]. PMID- 12774743 TI - [The new national postgraduate residency program and its relation to the general surgery board in Japan]. AB - The new national postgraduate residency program has been proposed to start from 2004 after long-term discussion. This new program consists of 2 years super rotating program with focusing onto the primary care. It requires basic clinical training in surgery for 3 months in the first year and extra 6 month can be used for optional period. On the other hand, the new general surgical board system has started from 2002 requiring 5 years training. This period is supposed to include the first 2 years postgraduate residency program. Also, a large number of postgraduate residents move from the University hospitals because the regulation sets the number of residents according to the hospital beds. There are various tasks and problems being proposed for these abrupt changes in the training system of doctors and especially for surgeons. This article summarized the interrelationship and problems with discussion. PMID- 12774744 TI - [Supporting system for regional medical liaison and role of a central hospital]. AB - The current status and future development of the supporting system for regional medical liaison and a role of the central hospital in the network were outlined. One of such supportive systems for regional medical network would be tele medicine or tele-mentoring that include radiological and pathological diagnoses in distance, tele-surgery, and tele-education. Most of these systems are facilitated in the universities and affiliated hospitals and generally need high cost communication equipment. Another approach is the information sharing system through the modern telecommunication network. Electronic patient record (EPR) systems are the key to achieving this and currently active in several areas. Since the recent progress in information technology (IT) is astonishing, community-based EPR systems are practical with the capability of clinical information exchange between different institutions and even with patients. The role of a central hospital in these systems must be capacious. Management and continuous operation of the system would be the most important affairs. For extending these supporting systems to the ones working in a broader area, the establishment of a "one ID for one patient" system is crucial. Strict security management of the data base and legal institution for distant medical practice still remain as the future tasks. PMID- 12774745 TI - [Changes in the surgical health care system and future aspect of surgery]. AB - Since universal coverage of health insurance of Japan was introduced in 1961. Japan's health care system has become one of the few comprehensive systems from the perspective of national welfare. In 2002, as the gross national medical expenditure (GNME) increased to 30 trillion yen, the Japanese Government employed a policy of medical cost containment. In Japan, the ratio of GNME to the gross national product (GNP) is not so high in the world, and it ranks 6th among developed countries (OECD-Health Data, 2002). During the past 38 years from the scientific viewpoint the Joint Committee of Social Insurance established by the Multidisciplinary Group of Surgical Associations has evaluated the rationalization of surgical fee (include doctor's fee). If our system is employed, more fair surgical care may be provided in the future. PMID- 12774746 TI - [The relationship between surgical and its subspecialty boards in Japan]. AB - This article deals with a 7-year endeavor to reform the board certification system of the surgical specialty and its subspecialties. The most important lesson learned is that the societies running the board must work not for the societies but for trainees and patients. The new postgraduate surgical training program is an overlapping system composed of a general surgery program of 5 years and optional subspecialty program of 7 years. There are four types of subspecialty program: cardiovascular; respiratory; gastroenterological; and pediatric surgery. The written examination for general surgery is taken 4 years after the start of training. Those who pass it and experience 350 general surgery cases within 5 years are eligible for oral examination by the surgery board. Those who pass the written examination for general surgery and experience a certain number of subspecialty surgeries within 7 years after the start of training are eligible for the oral and/or written examinations for the subspecialty board. The surgical societies are responsible for constantly improving the quality of the surgical training programs and qualification systems so that board-certified surgeons are accepted and treated as true surgical specialists in this country. PMID- 12774747 TI - [Introduction of new payment system (DRG/PPS) into university hospitals (2)]. PMID- 12774748 TI - [Medical malpractice, past, today and future]. PMID- 12774749 TI - [Considerations on the health care fee revision in the pediatric cardiovascular surgery]. PMID- 12774750 TI - The Drag King Anthology. Introduction. PMID- 12774751 TI - Posterior leukoencephalopathy in association with the tumour lysis syndrome in acute lymphoblastic leukaemia--a case with clinicopathological correlation. AB - We report a case and autopsy findings of posterior leukoencephalopathy (PL) developing during induction chemotherapy for B-cell acute lymphoblastic leukaemia (B-ALL) complicated by tumour lysis syndrome. PL may present with seizures, headache, altered mental status and occipital blindness, associated with transient parieto-occipital abnormalities on neuro-imaging studies. Precipitants include immunosuppressive agents, renal insufficiency, hypertension and fluid retention. It has also been reported in association with pre-eclamptic and eclamptic states, nephrotic syndrome and following liver and bone marrow transplantation. Only rare cases of PL developing during treatment for haematological malignancy have been reported and to our knowledge it has not been previously reported in association with tumour lysis syndrome. Since the condition is generally regarded as being fully reversible few autopsy findings have been reported. PMID- 12774752 TI - [Quantitative evaluation of veterinary assistants Voorjaarsdagen 2002]. PMID- 12774753 TI - ["Back to the future"]. PMID- 12774754 TI - [High volume hospital. The connection between number of cases and outcome quality in surgery]. PMID- 12774755 TI - [Psychotropic agents. 6/7 Mood altering agents]. PMID- 12774756 TI - [External comparative quality assurance in TEP in coxarthrosis. Results of 2001 German evaluation]. PMID- 12774757 TI - [Supplementary data on the "External comparative quality assurance in appendectomy. Results of 2001 German evaluation" article from the 2001 German Quality Report]. PMID- 12774758 TI - [DRG practices: code not found--what now? (Inpatient observation, vacuum sealing)]. PMID- 12774759 TI - [Competition and rational medicine. Specialist in a future-orientated integrated health care center]. PMID- 12774761 TI - [Management of burn victims in Yemen via Hammer Forum Initiative is decisively better]. PMID- 12774763 TI - [Wolfgang Muller-Osten Medical Academy]. PMID- 12774764 TI - [84th German Radiology Congress. 28-31 May 2003, Wiesbaden, Germany. Abstracts]. PMID- 12774765 TI - Digestive Disease Week and the 104th Annual Meeting of the American Gastroenterological Association. May 17-22, 2003. Orlando, Florida, USA. Abstracts. PMID- 12774767 TI - European Society of Paediatric Radiology 40th Annual Meeting and 26th Postgraduate Course. Genoa, Italy, June 2-6, 2003. Abstracts. PMID- 12774766 TI - Abstracts of the 38th Meeting of the Canadian Congress of Neurological Sciences. June 17-21, 2003, Quebec City, Canada. PMID- 12774768 TI - A bacterial enzyme to treat celiac disease? PMID- 12774770 TI - Questioning care. PMID- 12774771 TI - Mentoring. PMID- 12774769 TI - Enalapril accelerates remodeling of the renal interstitium after release of unilateral ureteral obstruction in rats. AB - Complete ureteral obstruction in rats rapidly leads to renal interstitial expansion and fibrosis and this process is ameliorated by concomitant angiotensin converting enzyme inhibition (ACEI). However, models of intervention initiated after unilateral ureteral obstruction (UUO) release may be more analogous to human obstructive renal disease where treatment could more reasonably follow the discovery of obstructive uropathy as compared to models where treatment is initiated at the time of experimentally induced obstruction. We studied interstitial changes in rats before and after release of UUO and examined the effect of ACEI with 200mg/L of enalapril (E) in the drinking water on these changes. Rats were sacrificed after 10 (n=10) and 20 (n=10) days (D) of UUO or 10D after release of 10D of UUO (n=18). Eleven rats received E for 10D after UUO release. Cortical interstitial volume fraction [Vv(I/C)] measured by point counting was increased at 10D (0.32 +/- 0.05) and 20D (0.41 +/- 0.05) of UUO compared to contralateral and sham-operated kidneys (both 0.05 +/- 0.01, ANOVA, p <0.001). Vv(I/C) 10D after release from 10D of UUO (0.26 +/- 0.04) was lower than that of 10D of UUO (p<0.05) and much lower than those with 20D of UUO (p<0.001). However, rats treated with E from the time of UUO release had lower Vv(I/C) (0.21 +/- 0.07) than UUO released E untreated rats (p<0.05). Release of UUO initiates regression of interstitial expansion in rats. ACEI with enalapril significantly accelerates reversal of interstitial expansion after UUO release. This could have important implications for treatment of obstructive nephropathy in humans. PMID- 12774772 TI - More on mentoring. PMID- 12774773 TI - Bone gun question. PMID- 12774774 TI - Renal involvement in Anderson-Fabry disease. AB - Anderson-Fabry disease (AFd) is a rare X-linked lisosomal storage disorder of glycosphingolipid (GL) metabolism, caused by a deficiency of the activity of alpha-galactosidase A (alpha-gal A). The progressive accumulation of GL in tissues results in the clinical manifestations of the disease, that are more evident in hemizygous males, and include characteristic skin lesions (angiokeratomas), neurological symptoms (acroparesthesia), ocular features (cornea verticillata), cardiac involvement (left ventricular enlargement, conduction abnormalities), cerebrovascular manifestations (thromboses, hemorrhage, etc.), and kidney involvement with progression to end-stage renal failure (ESRF). ESRF is a common manifestation in hemizygous males (3rd-5th decade) and death occurs around the 5th decade of life because of severe cardiac and/or cerebrovascular complications. Heterozygous females have an attenuated form of this systemic disease. In the kidney, accumulation of GL occurs in the endothelial cells of every vessel, in the epithelial cells of every tubular segment, and in all kinds of glomerular cells. The broad spectrum of renal lesions is a pathophysiological continuum with progressive impairment in the renal function related to continuous intracellular deposition of GL. Electron microscopic study of renal biopsies shows typical osmiophilic inclusion bodies in the cytoplasm of all kind of renal cells, characterized by concentric lamellation of clear and dark layers (35-50 A of periodicity). ESRF is treated by dialysis and kidney transplantation: neither treatment modifies the progression of the cardiovascular and cerebrovascular lesions due to progressive GL deposition. The outcome of kidney transplantation seems to be similar to that found in other non diabetic patients, but the survival rate on dialysis is lower than in patients with other causes of ESRF. Nowadays, treatment with enzyme replacement infusion with purified alpha-Gal A, produced by a genetically engineered human cell line or Chinese hamster ovocytes, seems to be effective and safe. PMID- 12774776 TI - Gene patent policy: does issuing gene patents accord with the purposes of the U.S. patent system? PMID- 12774775 TI - Preserving the right to choose: a minor's right to confidential reproductive health care. PMID- 12774777 TI - Moore revisited: state-sponsored biotechnological research and the takings clause. PMID- 12774778 TI - St Jude researchers use DNA chips to determine how leukemia cells responds to different drug treatments. PMID- 12774779 TI - Enforcement problems arising from conflicting views of living wills in the legal, medical and patient communities. PMID- 12774780 TI - European Stroke Services: From Evidence to Practice. Proceedings of the satellite symposium to the 11th European Stroke Conference. Geneva, Switzerland, May 29 June 1, 2002. PMID- 12774781 TI - Involuntary civil commitment and the new mental health bill. PMID- 12774783 TI - Tardive dyskinesia: legal issues and consent. PMID- 12774782 TI - Are medical ethics different from legal ethics? PMID- 12774784 TI - A code of ethics for social work and social care research. PMID- 12774785 TI - Genotype-dependent increase in plasma ACE activity after CABG is prevented by ACE inhibition. PMID- 12774786 TI - Worried? HIPAA privacy regs should change little. PMID- 12774787 TI - Religious beliefs and teenage refusal of medical treatment. PMID- 12774788 TI - Interleukin is as interleukin does. PMID- 12774789 TI - Adolescent autonomy, detention for medical treatment and Re C. PMID- 12774790 TI - Heroin addicts to receive CPR training and Narcan. PMID- 12774791 TI - Stenberg v. Carhart: women retain their right to choose. PMID- 12774792 TI - Use of stem cells in biotechnological research. PMID- 12774793 TI - Stroke prevention by the practitioner. PMID- 12774794 TI - Clinical quiz. Secondary carnitine deficiency due to celiac disease. PMID- 12774795 TI - [Treatment outcome with anti-HIV agent Tipranavir]. PMID- 12774796 TI - [96-week treatment outcome confirms long term efficacy of tenofovir DF]. PMID- 12774797 TI - [Decrease of HIV-1 virus load by nevirapin/efavirenz]. PMID- 12774798 TI - [Four years without resistance--newer standard in HIV therapy]. PMID- 12774799 TI - [ACTG-study 5095: superiority of efavirenz containing drug scheme]. PMID- 12774800 TI - [HIV-1 fusion inhibitor improves treatment in HIV-1-resistant patients]. PMID- 12774801 TI - Natural history of necrotizing pancreatitis. PMID- 12774802 TI - RE: Musculoskeletal injuries among ERCP endoscopists in Canada. Can J Gastroenterol 2002;16(6):369-374. PMID- 12774803 TI - Management of infected necrosis secondary to acute pancreatitis: a balanced role for minimal access techniques. PMID- 12774804 TI - The diagnosis is anencephaly and the parents ask about organ donation: now what? A guide for hospital counsel and ethics committees. PMID- 12774805 TI - Health care surrogate statutes: ethics pitfalls threaten the interests of incompetent patients. PMID- 12774806 TI - Recent developments in Kansas bioethics law: the Kansas Prevention of Assisted Suicide Act. PMID- 12774807 TI - Inappropriate advertising of dietary supplements. PMID- 12774809 TI - Inappropriate advertising of dietary supplements. PMID- 12774811 TI - A primer on posthumous conception and related issues of assisted reproduction. PMID- 12774812 TI - Procreative torts: enhancing the common-law protection for reproductive autonomy. AB - Roe v. Wade's twenty-fifth anniversary is likely to herald widespread scholarly commentary on the decision's continued vitality and the future of abortion in the United States. However, if such commentary focuses solely upon the constitutional dimensions and political aspects of a woman's right to privacy, an important dimension of this right will be overlooked. Few commentators have considered the extent to which tort law safeguards a woman's interest in reproductive autonomy. In this article, Professor Northern argues that the interest in reproductive autonomy has not yet received the full protection to which it is entitled and that tort law is poised to evolve distinct causes of action for the interference with procreative autonomy interests. Professor Northern begins with an overview of the medical and psychological literature on abortion-related risks. She goes on to discuss current trends in abortion malpractice litigation. The author then reviews the three basic types of malpractice causes of action--battery, negligence, and lack of informed consent--and explores their application to abortion malpractice claims. The focus of the article then shifts to the development of specialized procreative torts, and Professor Northern contends that courts should go beyond previous decisions to redress any substantial interference with procreative autonomy. Finally, the author asserts that legislative alternatives to the common-law development of procreative torts, such as right-to-know statutes, are less protective of women's interests. Professor Northern concludes that tort law could and should be used to more fully protect women's interests in procreative autonomy. PMID- 12774813 TI - [Reliability in diagnosis and therapy of breast cancer. First evidence-based guideline for breast cancer management on the internet]. PMID- 12774815 TI - The Aerosol Society Drug Delivery to the Lungs XIII. London, United Kingdom, December 12-13, 2002. Abstracts. PMID- 12774814 TI - Nephropathy induced by contrast medium. PMID- 12774816 TI - Nephropathy induced by contrast medium. PMID- 12774817 TI - Human cloning: a choice or an echo? PMID- 12774818 TI - Banning partial-birth abortion: drafting a constitutionally acceptable statute. PMID- 12774819 TI - Is the match illegal? PMID- 12774820 TI - Is the match illegal? PMID- 12774821 TI - Is the match illegal? PMID- 12774822 TI - Is the match illegal? PMID- 12774823 TI - Readability of informed-consent forms. PMID- 12774824 TI - The hepatobiliary-like excretory function of the placenta. A review. AB - In the adult, several endogenous compounds, such as bile acids and biliary pigments, as well as many xenobiotics are mainly biotransformed and eliminated by the hepatobiliary system. However, because this function is immature in the foetus, this role is carried out by the placenta during the intrauterine life. This review describes current knowledge of the trophoblastic machinery responsible for this function, which includes transport and metabolic processes, similar in part to those existing in the mature liver. Because many of the studies reviewed here were conducted on human or rat near-term placentae, two aspects should be borne in mind: (i) although both types of placenta are haemochorial, profound species-specific differences at the structural, molecular and functional levels do exist, and (ii) the placenta is an organ undergoing continuous developmental changes, including its hepatobiliary-like excretory function. PMID- 12774825 TI - Airway stenting. PMID- 12774826 TI - "Filing the gaps": Canadian voluntary nurses, the 1917 Halifax explosion, and the influenza epidemic of 1918. AB - During the Great War, St. John Ambulance established a reserve of approximately 2,000 casually trained nursing volunteers as Voluntary Aid Detachment nurses, or VADs, who assisted in military hospitals at home and overseas. During the 1917 Halifax explosion, and the 1918 influenza epidemic, VADs provided substantial assistance to qualified medical and nursing personnel. This paper considers their record during these specific events, arguing that despite the potential for VADs to undermine the status and job security of qualified graduate nurses, they proved instead both a valuable asset and a positive influence for the future development of Canadian nursing. PMID- 12774827 TI - Bandage battles: the American Red Cross and the politics of emergency response on the eve of World War I. AB - A century ago, first-aid instruction enjoyed sudden popularity on both sides of the Atlantic. In Britain and the Dominions, including Canada, classes were organised by the St. John Ambulance Association, while on the European continent several sophisticated national and municipal lifesaving associations rapidly outstripped the St. John model. In this article, which concentrates on hitherto neglected developments in the United States, seeks to establish similarities and differences within a comparative framework. Voluntarism, militarism, and modernity all receive attention, as does the question of bourgeois anxiety, seen by Roger Cooter as an important factor in creating "the moment of the accident". PMID- 12774828 TI - [Nephrotic syndrome (in the adult)]. PMID- 12774829 TI - Lesioning of spatial memory in mice treated with agroclavin. AB - This report presents results obtained from studies of the mnemotropic activity of the ergot alkaloid agroclavin. The effects of this substance were studied at concentrations of 1, 10, 25, 50, and 200 microg/kg on the learning and retention of a spatial habit in a Morris water maze. These doses of agroclavin had no effect on the process of formation of the spatial habit, but sharply disturbed retention of the habit. A long-lasting effect of memory trace disturbance was seen over a period of 48 h after dosage. On retraining, animals showed no delayed effects with agroclavin on learning a new spatial habit. The possible mechanisms for the effect of agroclavin on memory are discussed. PMID- 12774830 TI - Dynamics of intracellular dopamine contents in the rat brain during the formation of conditioned contextual fear and extinction of an acoustic startle reaction. AB - Extracellular dopamine contents in the caudate nucleus, nucleus accumbens, and prefrontal cortex of the rat brain were measured during two sessions of extinction of an acoustic startle reaction--each consisting of ten sound stimuli, the two sessions separated by 24 h--with simultaneous recording of freezing behavior. The results demonstrated a decrease in extracellular dopamine levels in the caudate nucleus and an increase in the nucleus accumbens during both sessions of extinction, with return to initial immediately after sessions ended. During the second session, the amplitude of startle responses and the magnitude of changes in dopamine levels in both structures were significantly smaller than during the first session. Between the sessions, dopamine levels in the caudate nucleus remained constant, while those in the nucleus accumbens decreased. The prefrontal cortex showed increases in dopamine levels during both sessions of extinction, as well as between the two sessions. The amplitude of the startle reaction was found to correlate with dopamine levels in the prefrontal cortex after the end of the corresponding extinction session and with the dopamine level before the start of the second session. The freezing time before the start of sound stimulation in the second session, this being a measure of conditioned fear, correlated with the dopamine level in the caudate nucleus on the training day and with the dopamine level in the nucleus accumbens before the start of the second session. The role of the dopaminergic system in the mechanisms forming and realizing the various components of defensive behavior are discussed. PMID- 12774831 TI - NMDA-dependent and NMDA-independent neural processes in the bicucculline disinhibited motor cortex of the cat during the acquisition and reproduction of a conditioned paw-on-support placing reflex. AB - Neuron activity was recorded in the motor area of the cat cortex during acquisition of an operant conditioned reflex consisting of placing the forepaw on a support in conditions of local disinhibition by spontaneous diffusion of the GABA(A) receptor blocker bicucculline from the recording micropipette. The conditioned signal was electrical stimulation of the parietal cortex with a train of 3-5 impulses. Addition of 2-amino-5-phosphopentanoic acid (APV), an NMDA glutamate receptor blocker, led to disappearance of the secondary excitatory components (in the poststimulus interval 30-120 msec) from neuronal responses in the disinhibited cortex both to the "indifferent" (before training) and the conditioned stimulation of the parietal cortex, while excitatory reactions associated with elevation and placing of the paw on the support showed no significant change in the presence of APV. Acquisition of the operant conditioned reflex was accompanied by an increase in the amplitude (p < 0.006) and duration (p < 0.00002) of secondary responses and decreases in their latent periods (p < 0.00002). In some cases--in fixed conditioned reflexes--secondary responses to conditioned stimulation in the disinhibited cortex were transformed into trains of epileptiform discharges. The hypothesis that changes in neuronal reactions in the disinhibited cortex during learning are based on increases in the efficiency of horizontal (collateral) connections between pyramidal neurons in layers II and III of the cortex is discussed. PMID- 12774832 TI - Is transfer of acquired coordination of head and forepaw movements possible in dogs? AB - Dogs were trained to tonic elevation of the forepaw and to use a lever to lift and maintain in position a food-containing cup during eating, this being accompanied by inclination of the head towards the feeder. In the conditions used here, the pretraining situation was that dogs would elevate the paw with an anticipatory upward movement of the lowered head; when the head tilted to the feeder, the paw flexed. The effect of special training, in which the initial coordination of the head and paw movements were remodeled, was that the animals maintained the paw elevated with the head in the lowered position. Dogs trained to perform the operant response with one paw did not transfer the acquired reaction when the "working" paw was changed. After the first training, the initial coordination was changed only between movements of the head and the "working" limb, but not between head movements and the non-trained paw. Remodeling of the initial movement coordination of the head with the second paw also occurred only as a result of the learning process. PMID- 12774833 TI - Analysis of cyclic adenosine-3',5'-monophosphate levels in structures of the "informational" and "motivational" systems of the rat brain during acquisition of a conditioned active avoidance reaction. AB - Cyclic adenosine-3'5'-monophosphate (cAMP) levels in structures of the "informational" and "motivational" systems of the brain were measured during acquisition of a conditioned two-sided active avoidance reflex in rats. cAMP levels were measured in three groups of animals--intact animals, trained animals, and an active control group (given uncombined presentations of the conditioned (light) and unconditioned (electric shock) stimuli)--immediately after reproduction of the acquired reflex. Significant accumulation of cAMP levels in brain structures was seen in animals of the active control group in the hypothalamus and in trained animals in the left and right hippocampus and the right frontal cortex. Positive correlations were found between cAMP levels in symmetrical parts of the frontal cortex, amygdala, and hypothalamus in animals of all study groups. In addition, active control rats and trained rats showed interhemisphere and intrahemisphere correlations between cAMP levels in brain macrostructure, whose patterns were specific for each group. The pattern of correlations observed here is assessed from the point of view of the role of the "informational" and "motivational" structures in the organization of adaptive behavior. PMID- 12774834 TI - Prototype symbolization in hooded crows. AB - Studies were undertaken to determine whether four crows previously trained to an image-based abstract selection rule could establish a relationship between the number of elements in a group and initially indifferent symbols (arabic numerals) and operate with these symbols, i.e., studies addressed the ability of these birds to symbolize. Unlike other similar studies, there was no use of special development of associative connections between the symbols and the corresponding arrays of elements, but conditions were created in which birds could observe these relationships on the basis of comparison with previously obtained information. Demonstration series were performed for this purpose, in which correct solutions by the crow resulted in receipt of a number of larvae corresponding to the number of elements in the array pictured or numeral imaged on the stimulus selected by the crow. Images belonged to the same category as the corresponding stimulus for selection (the second stimulus was another category): if the image was an array, then the corresponding picture for selection also bore an array, and vice versa. Crows could solve the task successfully by using an image-based selection rule. In test series, the image and both selection stimuli were initially from different categories: if the image was a numeral, then both selection stimuli were arrays, and vice versa. All four crows successfully coped with this task. Despite the absence of any similarity between the image and "correct" stimulus, they selected the array corresponding to the numeral and vice versa. It is suggested that the birds could achieve this result by comparing the information obtained during the presentation series--about the number of reinforcement units corresponding to each stimulus. Similar experiments showed that crows could use numbers to perform operations analogous to arithmetic addition. PMID- 12774835 TI - The effects of lesioning of the entopeduncular nucleus on the behavior of rats in simultaneous discrimination and open field conditions. PMID- 12774836 TI - Mental disorders in patients with malignant tumors of the maxillofacial region and larynx. PMID- 12774837 TI - Neuronal organization of the ventral anterior and ventral lateral nuclei of the human thalamus. PMID- 12774838 TI - Ischemic damage to neuron ultrastructure in organotypic cultures of hippocampal tissues. PMID- 12774839 TI - Structural rearrangements in the gray matter of the spinal cord after gravitational overloading. PMID- 12774840 TI - Morphofunctional changes in spinal cord tissues and spinal ganglia after epidural administration of clofelin. PMID- 12774842 TI - Organization of working memory processes in monkeys: the effects of a dopamine receptor agonist. AB - The effects of the dopamine receptor agonist agroclavin on cognitive processes associated with mechanisms of visual recognition and long-term and working (short term) memory were studied in delayed visual differentiation and delayed spatial selection tasks in monkeys (rhesus macaques). Measurements made before and after p.o. pharmacological treatment with this agent were used to identify the p.o. dose (5 mg/kg) inducing a significant effect. The psychotropic effect of agroclavin, which induced cognitive dysfunction, was present in all the monkeys studied to one extent or another. Behavioral criteria were: the probability of correct solutions of the visual differentiation task, the probability of refusals to resolve the task, and the time taken for correct motor responses. Despite individual differences in these behavioral characteristics in monkeys, significant changes due to agroclavin were consistently evident in all animals. There was a reduction in the probability of correct solutions, due to worsening of the characteristics of short-term memory; most monkeys showed increases in the numbers of refusals to solve tasks and increases in the time for correct motor responses during these solutions. In fact, all monkeys showed no increases in the number of erroneous solutions in visual differeniation and spatial selection tasks without delays, though in most cases there were increases in the time taken for correct motor reactions and the number of refusals to solve tasks. Data were obtained indicating that the effect of agroclavin was not uniform with respect to different types of visual information. The possible structural-functional organization of processes underlying working memory is discussed on the basis of the conclusion that the behavioral characteristics studied here reflect different components of cognitive processes realized by structures with different functional properties and different locations. PMID- 12774841 TI - The involvement of dopamine in strengthening cortical signals activating NMDA receptors in the striatum (a hypothetical mechanism). AB - A possible mechanism is proposed for the enhancement/weakening of those cortical signals in the cortex-basal ganglia-thalamus-cortex neural network which induce/do not induce opening of NMDA channels in the spiny neurons of the striatum and which can be regarded as "strong"/"weak" in terms of this measure. The mechanism is based on the modulatory influences of dopamine on changes in the efficiency of corticostriatal inputs. In the absence of dopamine, relative increases in the intensity of "strong" ("weak") cortical signals can lead to the induction of long-term potentiation (depression) of corticostriatal synapses. In this case, because of the differently directed influences on thalamic cells of signals passing via strionigral and striopallidal cells, "strong" signals at the output of the thalamus are weakened, while "weak" signals are strengthened. Activation of dopamine D1 (D2) receptors on strionigral (striopallidal) neurons may facilitate increases in the extent of long-term potentiation/depression (decreases in the extent of long-term potentiation/depression or induction of long-term potentiation/depression). The consequence of this is that "strong" signals at the output of the thalamus can be strengthened synergistically, while "weak" signals can be weakened synergistically. Background cortical signals evoking tonic release of dopamine in the striatum can decrease strengthening because of weakening of the modulatory influence of dopamine on the modification of corticostriatal synapses. PMID- 12774843 TI - Localization of corticoliberin receptors in the rat brain. AB - In situ hybridization was used to study the distribution of corticoliberin receptors of subtypes 1 and 2 (CL-R1 and CL-R2 respectively) in different structures of the rat brain. Levels of CL-R1 mRNA in the brain were significantly greater than levels of CL-R2 mRNA, and the most intense expression of the CL-R1 gene was seen in forebrain structures, especially various neocortical, archicortical, and paleocortical regions in the cerebellar cortex. In addition, significant levels of CL-R1 mRNA expression were noted in the red nucleus and the reticular nucleus of the tegmentum. Intense expression of CL-R2 mRNA was observed in structures of the olfactory system, corticomedial parts of the amygdala, fields CA1-CA4 of the hippocampus, the ventromedial hypothalamus, and several brain stem nuclei. Moderate levels of CL-R2 mRNA were seen in the dorsolateral neostriatum. These results provide evidence that corticoliberin receptors of both subtypes are widespread in the brain. The different patterns of expression of CL R1 and CL-R2 in the brain probably provide the basis for the functional specificity of action of corticoliberin in brain structures. PMID- 12774844 TI - Morphofunctional changes in the pineal gland during dynamic adaptation to hypothermia. AB - The effects of stress induced by hypothermia (+4 degrees C for 3 h) on the pathways of serotonin metabolism in the pineal gland and on its structure were studied in adult male Wistar rats. These experiments showed that the melatonin forming function of the epiphysis undergoes phasic changes during adaptation: there was a significant increase during the first 15 min, which was followed by gradual inhibition (to initial by 30 min) and then sharp suppression (at 3 h). Suppression of the functional activity of the pineal gland occurred because of exclusion of a proportion of pinealocytes from the process of active functioning. PMID- 12774845 TI - The neurite-stimulating activity of components of the salivary gland secretion of the medicinal leech in cultures of sensory neurons. AB - The effects of components of the salivary gland secretion (proteases and protease inhibitors) of the medicinal leech (Hirudo medicinalis) on the growth of neurites of sensory neurons from chick embryos (10-11 days old) were studied in organotypic cultures. Destabilase and high-molecular-weight bdellin B, (0.01, 0.02, 0.05, and 0.1 ng/ml), bdellastasin (0.02 and 0.05 ng/ml), and eglin C (0.1 ng/ml) had neurite-stimulating effects on day 3 of cultivation of spinal ganglia. Identification of the neurite-stimulating activity of these components of medicinal leech salivary gland secretions creates the basis for creating new therapeutic agents for the treatment of neurodegenerative diseases. PMID- 12774847 TI - Spatiotemporal dynamics of reentrant ventricular tachycardias in canine myocardial infarction: pharmacological modulation. AB - During the transition from a slow to rapid depolarization rhythm, rate-dependent sodium channel blockade develops progressively and increases from beat to beat under procainamide but more abruptly under lidocaine. We investigated the consequences of such differences on the dynamic course and stability of reentrant tachycardias at their onset. Procainamide and lidocaine were infused to equipotent plasma concentrations in canines with three-day-old myocardial infarction. We measured the activation times (ms) and maximum slopes of negative deflections in activation complexes (absolute value: /-dV/dt(max)/ in mV/ms) in 191 unipolar electrograms recorded from ischemically damaged subepicardial muscle during programmed stimulation inducing reentrant tachycardias. Procainamide caused a greater reduction in /-dV/dt(max)/ than did lidocaine in the responses to basic stimulation, and it favored the occurrence of cycle length prolongation at tachycardia onset as the /-dV/dt(max)/ decreased progressively in successive beats. This resulted in conduction block and tachycardia termination in three of eight preparations. In contrast, lidocaine caused a greater depression in / dV/dt(max)/ in response to closely coupled extrastimuli, but /-dV/dt(max)/ remained constant or even improved thereafter, and none of the tachycardias terminated spontaneously under lidocaine (n = 9). However, the reentrant circuits remained spatially unstable, and lidocaine favored the occurrence of cycle length dynamics displaying constant or decreasing trends. This study supports the notion that cycle length dynamics at tachycardia onset are determined by the properties of the reentrant substrate and their pharmacological modulation. PMID- 12774846 TI - The role of sex steroids in forming anxiety states in female mice. AB - Natural fluctuations in sex hormones during the ovarian cycle have enormous influences on ongoing psychological status in the female body. We report here studies of the effects of exogenous sex steroids on anxiety levels in female mice, as evaluated in the elevated cross maze test. Female NMRI mice were subjected to bilateral oophorectomy and one week later received s.c. injections of solvent (sesame oil, controls) or estradiol benzoate for 7 days, either alone or with an additional dose of progesterone on day 7. Elevated maze tests performed 6 h later showed that animals given progesterone had the highest levels of anxiety and the highest levels of grooming reactions as compared with the other groups. Immunohistochemical analysis of the distribution of progesterone receptors in different parts of the brain demonstrated significant increases in the numbers of immunopositive cells after injections of estradiol benzoate alone, with further increases after progesterone injections. Thus, the data obtained here suggest that the genomic effects of sex steroids are important, as they appear to be involved in non-sexual forms of behavior, particularly the level of anxiety. PMID- 12774848 TI - Bioavailability of anthocyanidin-3-glucosides following consumption of red wine and red grape juice. AB - Pharmacokinetic parameters and the bioavailability of several dietary anthocyanins following consumption of red wine and red grape juice were compared in nine healthy volunteers. They were given a single oral dose of either 400 mL of red wine (279.6 mg total anthocyanins) or 400 mL of red grape juice (283.5 mg total anthocyanins). Within 7 h, the urinary excretion of total anthocyanins was 0.23 and 0.18% of the administered dose following red grape juice and red wine ingestion, respectively. Pharmacokinetic parameters derived from plasma and urine concentrations exhibited higher variability after ingestion of red grape juice. Compared to red grape juice anthocyanins, the relative bioavailability of red wine anthocyanins was calculated to be 65.7, 61.3, 61.9, 291.5, 57.1, and 76.3% for the glucosides of cyanidin, delphinidin, malvidin, peonidin, petunidin, and its sum (referred to as total anthocyanins), respectively. Bioequivalence was established for none of the anthocyanins. On a low level, urinary excretion of anthocyanins was fast, and the excretion rates seem to exhibit monoexponential characteristics over time after ingestion of both red grape juice and red wine. Due to low bioavailability, any significant contribution of anthocyanins to health protecting properties of red wine or red grape juice seems questionable, but the clinical relevance of these findings awaits further investigation. PMID- 12774849 TI - Effects of cannabinoids on endogenous K+ and Ca2+ currents in HEK293 cells. AB - Effects of cannabinoids on endogenous potassium and calcium currents in HEK293 cells were studied using the whole-cell variant of the patch-clamp technique. The cannabinoid agonists WIN 55,212-2, methanandamide, and anandamide (1 microM) decreased the calcium current by 53.1 +/- 2.6, 47.5 +/- 1.2, and 38.8 +/- 3.1%, respectively, after transfection of human CB1 cannabinoid receptor (hCB1) cDNA into HEK293 cells. The delayed rectifier-like current was not changed after application of these agonists, but the inward rectifier was increased by 94.0 +/- 3.6, 83.7 +/- 5.1, and 63.0 +/- 2.5% after application of WIN 55,212-2, methanandamide, and anandamide, respectively. The effects of the cannabinoid antagonists (AM251, AM281, and AM630) on the inward rectifier and calcium currents were the opposite of those seen with cannabinoid agonists; thus, these compounds act as inverse agonists in this preparation. These results suggest that endogenous inward rectifier and calcium currents are modulated by cannabinoids in HEK293 cells, and that some expressed receptors may be constitutively active. PMID- 12774851 TI - Effects of acid-induced esophagitis on esophageal smooth muscle. AB - Acid-induced esophagitis is associated with sustained longitudinal smooth muscle (LSM) contraction and consequent esophageal shortening. In addition, LSM strips from opossums with esophagitis are hyper-responsive, while the circular smooth muscle (CSM) contractility is impaired. To determine the origin of these changes, studies were performed on esophageal smooth muscle cells isolated from opossum esophagi perfused intraluminally on 3 consecutive days with either saline (control; n = 8) or HCl (n = 9). CSM and LSM cells, obtained by enzymatic digestion, were exposed to various concentrations of carbachol (CCh) and fixed. CCh induced concentration-dependent contraction of both LSM and CSM cells. CCh induced LSM cell contraction was not different between control and esophagitis animals; however, there was marked attenuation in the CCh-induced contraction of CSM cells from esophagitis animals. Morphological studies revealed significant hypertrophy of the CSM cells. These findings suggest that impaired CSM contractility can be attributed at least in part to alterations to the CSM cell itself. In contrast, hyper-contractility demonstrated in LSM strips is likely related to factors in the surrounding tissue. PMID- 12774850 TI - Prostaglandin-independent effects of aspirin on cell cycle and putrescine synthesis in human colon carcinoma cells. AB - Aspirin consumption has been reported to be able to reduce colorectal cancer risk in humans and in animal models of colon carcinogenesis. Although the mechanism involved in such an effect is not yet clear, both prostaglandin-dependent and independent effects have been proposed. Using HT-29 Glc(-/+)cells, which originate from a human colon adenocarcinoma, we demonstrated in this study a dose dependent effect of millimolar concentration of aspirin on cell growth that was concomitant with a rapid accumulation of the cells in the G0/G1 phase, followed by an accumulation in the G2/M phase and by a minor increase in the proportion of cells undergoing nuclear condensation. Cell membrane integrity and cell release into the culture medium were not affected by this treatment. The aspirin effects were apparently unrelated to prostaglandin biosynthesis inhibition, since although these cells were found to express high levels of cyclooxygenase 1 (COX 1) and low levels of COX-2 proteins, they did not produce any measurable net amounts of prostaglandins, based on both utilization of radiolabelled arachidonic acid and the radioimmunoassay of prostaglandins E2 and F2 alpha. In contrast, we identified polyamine biosynthesis as a cellular target of aspirin, since the treatment of HT-29 Glc(-/+) cells with aspirin reduced the flux of L-ornithine through ornithine decarboxylase, an effect that could not be explained by an acute action of the drug on the ornithine decarboxylase catalytic activity. Since polyamine biosynthesis is strictly necessary for HT-29 cell growth, our data suggest that reduced flux through ornithine decarboxylase may participate in the antiproliferative activity of aspirin towards colonic tumoral cells. It is concluded that in HT-29 Glc(-/+) cells that are not functional for prostaglandin production, aspirin can affect cell growth, cell cycle, and polyamine biosynthesis without affecting cell membrane integrity. PMID- 12774852 TI - The beta1-adrenoceptor site activated by CGP12177 varies in behavior according to the estrous cycle phase and stress. AB - The aim of this work was to assess whether stress and estrous cycle phases affected the beta1-adrenoceptor (beta1-AR) site activated by CGP12177 in the right atria of rats. The chronotropic response to CGP12177 in the absence or presence of antagonists was determined in atria from rats submitted to one daily foot-shock session for 3 consecutive days. Blood was collected for hormonal assays. The pD2 for CGP12177 in atria from females was lower than in atria from males and was unaltered by stress or the estrous cycle. Propranolol (200 nM) or CGP20712A (3 microM) shifted the concentration-response curves to CGP12177 to the right in control and stressed estrus or control diestrus rats. Atria from stressed diestrus rats were resistant to blockade by propranolol or CGP20712A, indicating that the effect of beta-adrenoceptor antagonists on the response to CGP12177 is influenced by estrous cycle phases. The stress-induced increase in serum corticosterone levels was independent of the estrous cycle or gender, but the estradiol/progesterone ratio was affected differently in the two groups of female rats. In the diestrus group, serum estradiol levels decreased after the first foot-shock session and remained low until the day of sacrifice, whereas in the estrus group the serum levels of estradiol did not decrease after stress and peaked on the second day, which corresponded to proestrus. These data do not indicate whether there is a direct or indirect effect of stress hormones and (or) sex steroids on cardiac beta1-AR sensitivity. However, they do show that the classic and low-affinity binding sites of the beta1-AR are independently regulated and that the beta1-AR atypical site affinity for antagonists depends on the estrous cycle. PMID- 12774853 TI - Effect of exercise training on liver antioxidant status of deoxycorticosterone acetate salt induced hypertensive rats. AB - Several animal models have been developed to study the pathogenesis of hypertension. Deoxycorticosterone acetate (DOCA) salt induced hypertensive rats are adrenal models used to mimic human Conn's syndrome. Because previous studies showed a beneficial effect of chronic exercise (swimming) on the development of arterial hypertension in spontaneously hypertensive rats (which appears similar to human essential hypertension), we decided to evaluate the effects of swimming on DOCA-salt induced hypertension and liver antioxidant status. Therefore, the aim of this experiment was to study whether the swim training would improve hypertension and liver antioxidant status in DOCA-salt rats. DOCA-salt rats and control Sprague-Dawley rats were trained to swim 1 h/day, 5 days/week for 6 weeks and were sacrificed 48 h after the last exercise period. Systolic blood pressure was recorded before the sacrifice, and liver antioxidant status was evaluated in hepatic homogenates after the sacrifice. Swim exercise did not decrease systolic blood pressure in control and DOCA-salt rats but induced changes in liver activities of antioxidant enzymes, showing that exercise provoked liver oxidative stress in control and DOCA-salt rats. In comparison with our previous studies using spontaneously hypertensive rats, we conclude that the beneficial effects of chronic exercise on systolic blood pressure in rats are dependent on strain and the type of experimental hypertension. PMID- 12774854 TI - Chronic prenatal ethanol exposure-induced decrease of guinea pig hippocampal CA1 pyramidal cell and cerebellar Purkinje cell density. AB - The brain is a key target of ethanol teratogenicity, in which ethanol can produce neurodegeneration in selected areas, including the hippocampus and cerebellum. The research objective was to test the hypothesis that chronic prenatal ethanol exposure, via maternal ethanol administration, produces differential time course of decreased linear density of hippocampal CA1 pyramidal cells and cerebellar Purkinje cells. Timed pregnant guinea pigs received chronic oral administration of ethanol, isocaloric-sucrose/pair-feeding, or water throughout gestation (term, about gestational day (GD) 68), and the offspring were studied at GD 62 (near term fetus), postnatal day (PD) 1 (neonate), PD 5, and PD 12 (early postnatal life). Ethanol treatment, compared with isocaloric-sucrose/pair-feeding and water treatments, decreased brain, hippocampal, and cerebellar weights at GD 62, PD 1, PD 5, and PD 12. Hippocampal CA1 pyramidal cell linear density and cerebellar Purkinje cell linear density were unaffected at GD 62. Ethanol treatment produced 25, 30, and 30% decreases in linear density of hippocampal CA1 pyramidal cells at PD 1, PD 5, and PD 12, respectively, and a 30% decrease in linear density of cerebellar Purkinje cells at PD 12 only. At PD 5, Purkinje cell profile linear density remained unaffected; however, ethanol treatment appeared to increase linear density of apoptotic Purkinje cell nuclei, as determined by a modified TUNEL method. The data demonstrate that chronic prenatal ethanol exposure produces apparent differential time course of decreased linear density of hippocampal CA1 pyramidal cells and cerebellar Purkinje cells in the developing guinea pig. PMID- 12774855 TI - Effects of simvastatin and pravastatin on peroxidation of erythrocyte plasma membrane lipids in patients with type 2 hypercholesterolemia. AB - Since hypercholesterolemia directly modifies the composition of erythrocytes plasma membrane, the influence of statins on erythrocytes has been researched. The beneficial effects of statins on clinical events may involve mechanisms that modify endothelial dysfunction, plaque stability, thrombus formation and inflammatory responses. The aim of the study was to evaluate the hypolipemic efficacy and effects of pravastatin and simvastatin on erythrocyte membrane fluidity and damage of erythrocytes in patients with type 2 hypercholesterolemia in comparison with a control group of healthy subjects. The study involved 53 patients affected by type 2 hypercholesterolemia (mean age, 53.3 +/- 10.3) with initial total serum cholesterol (TC) levels > 250 mg/dL, LDL-cholesterol (LDL-C) levels > 170 mg/dL, and triglycerides (TG) levels < 400 mg/dL. The control group consisted of 30 healthy individuals (mean age 56.9 +/- 6.3). Statins were given for 12 weeks. The dosages for oral administration of simvastatin and pravastatin were 20 mg/day. Laboratory tests were carried out before and after 4 and 12 weeks of the pharmacological treatment. The damage to plasma membrane of erythrocytes was measured on the basis of lipid peroxidation. The fluidity of plasma membrane of erythrocytes was determined by electron paramagnetic resonance (EPR) spectroscopy, using two spin labels: 5-DSA and 16-DSA. The cholesterol level in the membrane of red blood cells was estimated. Simvastatin and pravastatin reduced the total cholesterol concentration and LDL-cholesterol in plasma, as well as the cholesterol concentration in erythrocytes membranes. Hypercholesterolemia induced changes in the basic properties of human erythrocyte plasma membrane, including its fluidity and the intensity of lipid peroxidation. These results indicate that the simvastatin and pravastatin therapy reverses the alteration in the erythrocyte plasma membrane properties. PMID- 12774856 TI - Working heart function in diabetes is not improved by spironolactone treatment. AB - Aldosterone antagonism has emerged as an important strategy for end-stage congestive heart failure. To evaluate the potential contribution of aldosterone towards the cardiac complications of diabetes, this study examined the effects of chronic aldosterone receptor blockade (with spironolactone) on isolated working heart function in streptozotocin (STZ) - induced diabetic rats. Wistar rats were divided into four groups: control, control spironolactone-treated, diabetic, and diabetic spironolactone-treated. Following chronic spironolactone treatment (8 weeks), cardiac function was assessed in terms of the rate of contraction (+dP/dT), rate of relaxation (-dP/dT), and left ventricular developed pressure (LVDP). Untreated diabetic rats exhibited marked cardiac dysfunction when compared with age matched controls (p < 0.001). Long-term spironolactone treatment did not improve these parameters. These data demonstrate the lack of beneficial effects of aldosterone receptor blockade on isolated working heart function in diabetes. PMID- 12774857 TI - A novel vasopressin peptide lowers blood pressure through decreases in cardiac output. AB - The changes in blood pressure, cardiac output, and total peripheral conductance evoked by the novel hypotensive arginine vasopressin (AVP) - like peptide, d(CH2)5[D-Tyr(Et)2,Arg3,Val4,Arg7,Eda9]AVP (HYPO-AVP), were recorded in conscious unrestrained Sprague-Dawley rats implanted with radiotelemetry pressure transducers and ultrasonic transit-time flowprobes. Intravenous infusions of 0.6, 1.0, 2.0, and 4.0 microg x kg(-1) x min(-1) of HYPO-AVP evoked dose-related decreases in blood pressure. At the lowest dose of 0.6 microg x kg(-1) x min(-1), the fall in blood pressure was associated with a small but significant increase in total peripheral conductance. Cardiac output was unchanged. In contrast, at the three higher doses of 1.0, 2.0, and 4.0 microg x kg(-1) x min(-1), the fall in blood pressure was related to a dramatic fall in cardiac output. Indeed, total peripheral conductance decreased, preventing blood pressure from falling further. These hemodynamic findings should help to direct future research into the mechanism of the putative hypotensive property of vasopressin, a property that attenuates the well established blood pressure elevating actions of the peptide. PMID- 12774858 TI - Who are the comorbid adolescents? Agreement between psychiatric diagnosis, youth, parent, and teacher report. AB - The investigators examined the rates of psychiatric comorbidity for externalizing and internalizing behavior problems, using semistructured diagnostic interview and parent, teacher, and youth report on the Achenbach checklists. The study also evaluated the effects of conjunctive, compensatory, and disjunctive data combination strategies. Using the same data and identical diagnostic thresholds, between 5 and 74% of 189 youths presenting to an outpatient clinic were identified as having comorbid internalizing and externalizing problems. Parent report and semistructured interview indicated the highest comorbidity rates. Despite good cross-source agreement (rs .29-.58), there was very little agreement about which specific youths presented with comorbid internalizing and externalizing problems (kappas .14-.40). Results also indicate that single DSM-IV disorders, such as bipolar disorder, can manifest "comorbid" patterns of behavior problems on checklists. PMID- 12774859 TI - Untangling developmental relations between depressed mood and delinquency in male adolescents. AB - Relations between depressed mood and delinquency were investigated in a longitudinal sample of 506 urban adolescent males across ages 13.5-17.5, while adjusting for common risk factors. Adolescents provided yearly reports of their delinquent activities and depressed mood, as well as reports of peer delinquency at age 13.5 (i.e., baseline). Primary caregivers and teachers provided reports of risk factors for depressed mood and delinquency such as aggressive behavior problems and low academic achievement. Two-level hierarchical generalized linear models of concurrent relations indicated that depressed mood predicted concurrent variety of delinquent acts, and more variety of delinquent acts predicted concurrent depressed mood, even after controlling common risk factors. Longitudinal analyses indicated that after controlling for common risk factors, depressed mood had a more robust effect on delinquency variety trajectories than delinquency variety had on depressed mood trajectories. Time-averaged depressed mood significantly predicted a more positive rate of change in delinquency variety across time. Baseline delinquency variety predicted baseline depressed mood and time-averaged delinquency variety predicted a more positive rate of change in depressed mood; however, both effects were marginally significant. Implications of the results for theory and intervention are discussed. PMID- 12774861 TI - Comorbidity between and within childhood externalizing and internalizing disorders: reflections and directions. AB - The term and concept of "comorbidity" has been mired in controversy, although there is little question that the existence of covariation among psychiatric diagnoses poses significant challenges to current models of psychiatric classification and diagnosis. The papers in this Special Section underscore a number of important issues relevant to the comorbidity between and within childhood externalizing and internalizing disorders, and illustrate both methodological and substantive reasons for such comorbidity. Weiss, Susser, and Catron's distinction among common, broad-band specific, and narrow-band specific features provides a helpful framework for understanding the comorbidity of childhood externalizing and internalizing disorders (B. Weiss, K. Susser, & T. Catron, 1998). Hierarchical models of psychopathology help to dissolve the distinction between "splitters" and "lumpers" and point to variables that may elucidate the etiology of externalizing and internalizing disorders. PMID- 12774860 TI - Differential risks of covarying and pure components in mother and teacher reports of externalizing and internalizing behavior across ages 5 to 14. AB - In a sample of 585 children assessed in kindergarten through 8th grade, we fit a confirmatory factor model to both mother- and teacher-reported symptoms on the Achenbach checklists (CBCL, TRF) and determined that a covariation factor of externalizing and internalizing behaviors existed, in addition to the pure-form factors of externalizing and internalizing for each reporter. In 3 structural equation models, between 8 and 67% of the variance in these 6 latent factors was accounted for by a set of antecedent child, sociocultural, parenting, and peer risk variables. Each of the 6 latent factors, taken 2 at a time, was predicted by a unique set of risk variables; however, there were some patterns that held for both mother- and teacher-report symptom factors: Child temperamental unadaptability and female gender were predictors of higher internalizing symptoms; child temperamental resistance to control, parental harsh punishment, male gender, low SES, and peer rejection were related to higher externalizing symptoms whereas child temperamental unadaptability was related to lower externalizing symptoms; and peer rejection and family stress were also related to the covarying, externalizing-plus-internalizing component of both mother and teacher reports. PMID- 12774862 TI - Comorbidity and child psychopathology: recommendations for the next decade. AB - This special section exemplifies and offers a number of important methodologic and conceptual advances that should provide investigators new tools for understanding comorbidity of child and adolescent psychopathology, including (a) the importance of making careful methodologic distinctions in how comorbidity is defined and operationalized, (b) specifying and justifying how data from different sources are combined, (c) teasing out the impact of potentially confounding risk factors that lead to symptom and syndrome overlaps, and (d) exploring the effects of time, timing, and order of disorder emergence on variable manifestations of comorbidity. These advances are much needed, but may still prove insufficient, given the daunting challenges in fully understanding comorbidity. Thus, future studies should be characterized by (a) more focused search for subgrouping factors and interactions related to the emergence of comorbidity, (b) careful exploration of setting- and/or informant-specific types of psychopathology, (c) development of studies that explore not just phenotypes and genotypes, but also environtypes and trajectory-types, (d) more discriminative use of information sources, including explicit efforts to reconcile (rather than combine) discrepant information, (e) clear descriptions and logical justification of when conjunctive, disjunctive, additive, and discriminative combinatorial approaches are used, (f) increased use of multidisciplinary research methods and teams, (g) increased application of multiple lines of evidence in comorbidity studies, (h) increased focus on understanding illness processes rather than just psychopathologic states, (i) development of creative new research designs, and (j) redrawing disorder boundaries when warranted. PMID- 12774863 TI - Motor timing deficits in community and clinical boys with hyperactive behavior: the effect of methylphenidate on motor timing. AB - In a previous paper we showed that community children with hyperactive behavior were more inconsistent than controls in the temporal organization of their motor output. In this study we investigated: (1) various aspects of motor timing processes in 13 clinically diagnosed boys with attention deficit hyperactivity disorder (ADHD) who were compared to 11 community boys with hyperactive behavior and to a control group and (2) the effect of methylphenidate on the motor timing processes in the clinical group with ADHD in a double blind, cross-over, medication-placebo design, including 4 weeks of medication. The clinical group with ADHD, like the community group with hyperactivity, showed greater variability in sensorimotor synchronization and in sensorimotor anticipation relative to controls. The clinical group was also impaired in time perception, which was spared in the community group with hyperactivity. The persistent, but not the acute dose, of methylphenidate reduced the variability of sensorimotor synchronization and anticipation, but had no effect on time perception. This study shows that motor timing functions are impaired in both clinical and community children with hyperactivity. It is the first study to show the effectiveness of persistent administration of methylphenidate on deficits in motor timing in ADHD children and extends the use of methylphenidate from the domain of attentional and inhibitory functions to the domain of executive motor timing. PMID- 12774864 TI - Selective inhibition in children with attention-deficit hyperactivity disorder off and on stimulant medication. AB - Selective inhibition requires discrimination between auditory signals and is assessed using a modification of the stop-signal task. Selective inhibition was assessed in a group of 59 clinic-referred, DSM-IV-diagnosed children with attention-deficit hyperactivity disorder (ADHD) and compared to that of a community sample of 59 children. Methylphenidate (MPH) effects on selective inhibition were assessed in a subset of the ADHD sample that participated in an acute, randomized, placebo-controlled, crossover trial with 3 fixed doses of MPH. Children with ADHD performed more poorly than controls on the majority of selective stop-signal task parameters: they exhibited more anticipatory (invalid) responses, with less accurate and more variable responses on the response execution task, as well as a slower selective inhibition process. MPH improved speed of both inhibition and response execution processes; it also reduced variability of response execution and decreased nonselective inhibition. On the one hand, findings are consistent with purported inhibition deficit in ADHD, but on the other hand, suggest that neither the impairment itself, nor MPH effects, were restricted to inhibition. PMID- 12774865 TI - Hostile attributional tendencies in maltreated children. AB - The hostile attributional tendencies of maltreated children in elementary school across key relationship figures (i.e., parents, teachers, and peers), the relation between children's hostile attributional tendencies and the frequency and severity of maltreatment, and the role of children's hostile attributions of their parents in mediating the relation between maltreatment and children's hostile attributions of unfamiliar peers were examined. The sample consisted of 44 maltreated and 56 nonmaltreated children (females = 51) of mixed ethnicity. Subjects were administered a 20-item measure of attributional processes. The results indicated that relative to nonmaltreated children, physically abused boys were more likely to attribute hostile intentions to a variety of relationship figures, including their parents, an unfamiliar teacher, their best friend, and unfamiliar peers. A positive relation was also found between the frequency of physical abuse and hostile attributional tendencies among males. Finally, support was found for the role of children's hostile attributions of their mothers in mediating the relation between physical abuse and children's hostile attributions of unfamiliar peers. The results support a link between physical abuse and hostile attributional tendencies in children in early elementary school. PMID- 12774866 TI - Amazed or appalled, apathy or action? PMID- 12774867 TI - Soaring cesarean section rates: a cause for alarm. PMID- 12774868 TI - Hormone replacement therapy. PMID- 12774870 TI - Prevention and treatment of vulvovaginal candidiasis using exogenous Lactobacillus. AB - OBJECTIVE: To review literature examining exogenous Lactobacillus therapy for vulvovaginal candidiasis and to discuss recommendations for clinical practice and future research. DATA SOURCES: Computerized searches on MEDLINE and CINAHL November 2000, September 2001, and March 2002, with search terms including Lactobacillus, acidophilus, Candida, and yeast infections. STUDY SELECTION: Relevant English-language articles from the past 10 years. Unique or seminal studies included where pertinent. DATA EXTRACTION AND SYNTHESIS: Data organized under the following headings: endogenous Lactobacillus, exogenous Lactobacillus, Candida, studies of intravaginal Lactobacillus therapy for vulvovaginal candidiasis, studies of oral Lactobacillus therapy for vulvovaginal candidiasis. CONCLUSIONS: Vaginally administered or orally ingested Lactobacillus is able to colonize the vaginal ecosystem. Controlled intervention studies regarding the effect of such colonization on vulvovaginal candidiasis are promising but few. These studies had small numbers of participants, were inconsistent in the form of Lactobacillus used, and reported conflicting results. Further randomized controlled trials involving large numbers of women are imperative. In the meantime, health care providers should discuss potential benefits with affected patients while clarifying the current lack of conclusive evidence. Without further research into currently available sources and brands of Lactobacillus and without governmental regulation of supplements and their contents, however, it is difficult to make recommendations regarding appropriate product choice. PMID- 12774871 TI - Deep tendon reflexes: the what, why, where, and how of tapping. AB - Deep tendon reflexes demonstrate the homeostasis between the cerebral cortex and the spinal cord. When these reflexes are disrupted, hyperreflexia (disease induced) or hyporeflexia/areflexia (drug induced) occurs. Although nurses perform deep tendon reflex assessments regularly, it is difficult to incorporate theoretical principles in these assessments because of scant medical literature, a lack of nursing research, and time constraints in nursing programs. These conditions usually result in one-on-one training, causing reduced consistency. A comprehensive examination assists the clinician to apply theoretical principles, develop expert technique, and serve as a catalyst for clinical research. PMID- 12774872 TI - Limited obstetric ultrasound examinations: competency and cost. AB - OBJECTIVE: To determine both the actual dollar cost and the amount of time required per nurse to establish competency in limited obstetric ultrasonography (LOBU). DESIGN: Descriptive. SETTING: A tertiary care setting. PARTICIPANTS: Registered nurses who were taught in LOBU. RESULTS: Nurses who attained competency in LOBU completed 12 hours of didactic education and a clinical practicum consisting of 6 to 9 hours and approximating 15 ultrasound scans. For five nurses to concurrently attain competency in LOBU, the cost per nurse was $1,037.55 (includes salaries and employee benefits). CONCLUSION: Registered nurses are able to acquire competency in LOBU at a reasonable cost, thus enhancing the ability of the professional nurse to deliver a fuller scope of services in an obstetric setting. For institutions that have limited access to individuals with this skill, nurses trained in LOBU may present a high-quality, cost-efficient solution to providing needed obstetric services. PMID- 12774873 TI - A comparison of pregnancy health practices of women in inner-city and small urban communities. AB - OBJECTIVE: To examine whether the relationships among pregnancy health practices, maternal-fetal attachment, and prenatal depression differ between women living in the inner city and women living in a smaller urban community. DESIGN: Cross sectional, descriptive survey. SETTING: Clinics in inner-city and small urban communities in the Midwest. PARTICIPANTS: Fifty-five pregnant women living in the inner city and 197 pregnant women living in a smaller urban setting. MAIN OUTCOME MEASURES: Health Practices Questionnaire, a self-report instrument that measures pregnancy health practices. RESULTS: Women living in the inner city reported lower levels of health practices than women in urban settings, but this was moderated by maternal-fetal attachment. Inner-city women with lower levels of maternal-fetal attachment had poorer health practices than inner-city women with higher levels of maternal-fetal attachment; there was no relationship for women residing in the small urban area. Depression was negatively associated with health practices for all women; however, there was no difference by residence. CONCLUSION: Maternal-fetal attachment contributed differently to health practices in women from the inner city compared with women from small urban communities. Nurses can use this information to help identify women at risk for poor pregnancy health practices. More research is needed to identify the interventions that most effectively improve health practices. PMID- 12774874 TI - The effect of ultra low dose epidural analgesia on newborn breastfeeding behaviors. AB - OBJECTIVE: To determine whether a difference in breastfeeding behaviors could be observed between newborns whose mothers received epidural analgesia for labor pain relief and those newborns whose mothers received no pain medication in labor. DESIGN: There were two groups of neonates in this study. One group was born to mothers who received epidural analgesia, and one group was born to mothers who received no pain medication for labor. Both groups were observed for initial breastfeeding behaviors using the Premature Infant Breastfeeding Behavior Scale following birth and at 24 hours. Central nervous system functioning in the newborn was measured with the Neurologic and Adaptive Capacity Score at 2 and 24 hours of age. SETTING: A large tertiary hospital in northeast Ohio. PARTICIPANTS: Fifty-six breastfeeding mother-newborn dyads. All mothers were healthy multiparae who gave birth vaginally to normal, full-term, healthy newborns. MAIN OUTCOME MEASURES: Newborns were observed for rooting, latch on, sucking, swallowing, activity state, and neurobehavior. RESULTS: There were no statistically significant differences in breastfeeding behaviors at birth or at 24 hours of age. CONCLUSION: A possible cause for the lack of significant results may have been the ultra low dose of bupivacaine and fentanyl used in this sample. PMID- 12774875 TI - Breast binding... is it all that it's wrapped up to be? AB - OBJECTIVE: To investigate the difference in breast symptoms between breast binding and support bra wearing in nonbreastfeeding postpartum mothers. DESIGN: A systematic replication of an earlier study by Bristol using a pre-experimental posttest design. SETTING: A private, for-profit hospital in a city in the south central region of the United States. PARTICIPANTS: Sixty nonbreastfeeding postpartum women who gave birth to viable newborns of singleton gestations, had an uncomplicated postpartum, and did not receive hormonal lactation suppressants. MAIN OUTCOME MEASURES: Postpartum breast engorgement, leakage, tenderness, and use of pain relief measures as measured by the Bristol Record of Symptoms. RESULTS: Analysis of the data revealed no significant difference relative to breast engorgement between the two groups during the first 10 postpartum days. However, the breast-binder group reported a greater degree of breast tenderness, breast leakage, and use of other pain relief measures. CONCLUSION: Breast binding should be discontinued as a method of lactation suppression and use of support bras encouraged. Future studies need to focus on comfort for nonbreastfeeding, postpartum mothers. PMID- 12774876 TI - Co-bedding versus single-bedding premature multiple-gestation infants in incubators. AB - OBJECTIVE: To compare the physiological stability and behavioral effects of co bedding with those of single-bedding premature multiple-gestation infants in incubators as well as the psychological effects on their parents. DESIGN: Prospective, randomized, repeated measure. PARTICIPANTS: Convenience sample of 16 infants and 8 parents in the co-bedded group, and 21 infants and 11 parents in the control group. INTERVENTIONS: Infants in the study group were co-bedded in incubators. MAIN OUTCOME MEASURES: Baseline and posttesting for parental state anxiety, maternal attachment, and parental satisfaction measures; infant sleep wake synchronicity; physiological measures; and stress cue measures during baseline and activity. MAIN RESULTS: Repeated measures 5 (time) x 2 (group) analysis of variance found significant differences in infant daily weight, feeding amount, and high-activity heart rate. There was no difference in parental state anxiety, maternal attachment, and parental satisfaction scores by group, except for higher baseline parental satisfaction scores in the co-bedded group. CONCLUSIONS: This research demonstrated the safety of co-bedding multiple gestation infants in incubators but did not find any significant clinical improvement in infant or parental outcomes with co-bedding. Neonatal intensive care unit providers should educate staff and parents about the potential benefits of co-bedding and consider developing policies and procedures for co-bedding in both incubators and cribs. Co-bedding of multiple-gestation infants may be provided as an adjunctive developmental care strategy if parents desire this intervention. PMID- 12774877 TI - The lived experience of nurses caring for newborns with sepsis. AB - OBJECTIVE: To describe the lived experience of nurses who care for newborns with sepsis. DESIGN: Phenomenology, a qualitative study using open-ended, tape recorded interviews, followed by a focus group discussion. The data were analyzed using Colaizzi's step-by-step procedure, resulting in an essential structure of the experience. SETTING: The nursery and neonatal intensive-care unit of a large community hospital in the southwestern United States. PARTICIPANTS: Eleven registered nurses who cared for newborns in both a transitional nursery and the neonatal intensive-care unit were interviewed individually or in a focus group. RESULTS: Three major themes were generated from the data: (a) "Dealing With Death and the Blessings of Life," (b) "Sepsis: The Cancer in the NICU," and (c) "Losing the Dream: Parents' Reactions." The nurses had feelings of helplessness and frustration while caring for these sick newborns. Nurses described the changes in newborns' condition as sepsis developed, the actions taken to reverse the downhill course, and the experiencing of the outcome. Nurses perceived that parents responded with overwhelming fear, guilt, and loss of control. CONCLUSIONS: The nurse caring for the infant with sepsis experiences many different emotions, reactions, and perceptions. These findings can assist nurses to have a better understanding of the role of the nurse and the emotional burden of working in the NICU. PMID- 12774878 TI - Decision making regarding multifetal reduction. AB - OBJECTIVE: To identify salient variables that influence decision making regarding multifetal reduction (MFR) and describe their effect on individuals over time. DESIGN: Prospective, exploratory, descriptive design, using qualitative and quantitative methods. SETTING: Midwestern tertiary care center. PARTICIPANTS: A convenience sample of 11 consecutive consenting couples with triplet or higher order pregnancies who elected to undergo MFR. METHODS: Semistructured audiotaped telephone interviews at three points: (a) 2 weeks postreduction, (b) 6 weeks postpartum, and (c) 6 months postpartum; a demographic and marital adjustment questionnaire. MAIN OUTCOME MEASURES: Themes identified by content analysis and compared via matrix analysis between males and females and at three points in time; trends in marital adjustment. RESULTS: Dominant variables influencing MFR decision making were risks associated with higher-order pregnancies and preservation of infants' and mothers' health. Most participants identified emotional issues, including moral and ethical dilemmas, as the most difficult aspect of reduction. Over time, participants reported feeling more positive about their decision; nonetheless, negative feelings emerged progressively. CONCLUSIONS: Risk aversion favored MFR decision making. Yet, both making and living with the decision were emotionally difficult for this sample. Interventions are needed to assist couples with this decision and its consequences. PMID- 12774879 TI - Annotated bibliography of recent National Institute of Nursing Research findings on women's health across the life span. PMID- 12774880 TI - An organizational guide for an effective developmental program in the NICU. AB - Many newborn intensive care units have put a great deal of energy and resources into their efforts to provide developmentally supportive care. Yet, without consistent leadership and clear accountabilities, developmental care will depend on the individual philosophy, or even the mood, of the health care professional at the bedside. Under these circumstances, infants and families experience an unpredictable and inconsistent quality of care. This article presents a guide to assist nurseries in structuring successful programs of developmentally supportive, family-centered care through well-described accountabilities that translate into consistent, high-quality care for infants and families in the NICU. PMID- 12774881 TI - Challenges in providing developmentally supportive care: a case presentation. AB - A developmentally supportive family-centered approach to care is extremely valuable for even the most critically ill infant. This case presentation of Martha, a severely small for gestational age twin born at 30 weeks gestation, highlights three major areas in which clinical care is enhanced through the integration of neurobehavioral observation: self-regulation, comfort, and family advocacy. The ethical principles of autonomy, beneficence, and integrity were relied upon to ensure her humane care. PMID- 12774882 TI - Sleeping beauties: the impact of sedation on neonatal development. AB - Sedatives are frequently administered in neonatal intensive care to induce sleep for diagnostic and radiology procedures, calm irritable infants, manage pain related agitation, and enhance ventilation. The pharmacology and side effects of sedatives commonly used with neonates will be reviewed and placed within the context of their potential effect on neonatal development. Alternative caregiving strategies to minimize or eliminate the need for sedation will be discussed. PMID- 12774883 TI - Factors influencing language development in preterm infants. AB - OBJECTIVE: To examine factors influencing preterm infant language development. DESIGN: Longitudinal. SETTING: Infants were seen for developmental follow-up at 7, 13, and 26 months corrected age in the school of nursing. PARTICIPANTS: The sample consisted of 43 mothers and their preterm infants who were below 2,000 g and 36 weeks gestation at birth. More than 88% of the sample were from lower social classes. Seventy-three percent of the sample was African American and 27% was White. MAIN OUTCOME MEASURES: Developmental outcome was assessed using the Bayley Scales of Infant Development, and language was assessed using the Reynell Developmental Language Scales. RESULTS: By 26 months corrected age, infant development was within the normal range. Expressive and receptive language was delayed an average of 3 to 5 months. Factors influencing language included length of hospital stay, birth weight, Apgar scores, infant irritability and state regulation at hospital discharge, and maternal sensitivity. CONCLUSION: Language development is delayed in preterm infants. Maternal sensitivity is positively associated with enhanced infant language. Nurses need to utilize opportunities to enhance sensitive mothering to optimize infant outcomes. PMID- 12774884 TI - Favourable influence of low molecular weight heparin in mitigating the peroxidative membrane damage induced by a cytotoxic agent and an atherogenic diet. AB - The present study is aimed to demonstrate the protective effect of a heparin derivative, low molecular weight heparin (LMWH) against erythrocyte membrane injury. Two models serve to induce membrane lipid peroxidative damage, namely a potent cytotoxic agent, adriamycin and a hypercholesterolemic atherogenic diet. Two groups of male Wistar rats (140 +/- 10 g) received a single intravenous injection of adriamycin (ADR, 7.5 mg/kg), while two other groups were fed an atherogenic diet comprising a supplementation of 4% cholesterol, 1% cholic acid and 0.5% thiouracil (CCT diet) for 2 weeks. For each of the above two groups, LMWH (Troparin; 300 microg/day per rat subcutaneously) treatment commenced on day 8 and continued for a week. One group was maintained as the normal control group, and another group that received only LMWH treatment was designated as the LMWH drug control group. Erythrocyte membrane was isolated and assayed for its cholesterol levels, lipid peroxidation and ATPases activity. The activities of antioxidant enzymes were assessed in the haemolysate. The findings of the study were that both adriamycin and the atherogenic diet produced elevated membrane cholesterol levels and lipid peroxidation. The membrane ATPases suffered loss in activity. Accentuated oxidative stress was marked by rise in the activities of antioxidant enzymes (SOD, catalase and GPx). LMWH intervention reverted these changes thereby normalizing the membrane composition and function. The membrane protective effect of LMWH is illuminated by this work. PMID- 12774885 TI - Acute lead exposure induces renal haeme oxygenase-1 and decreases urinary Na+ excretion. AB - The effects of acute lead exposure on renal function, lipid peroxidation and the expression of haeme oxygenase (HO) in rat kidney were determined. A single injection of lead acetate (50 mg Pb/kg) was given to rats. Changes in renal function, characterized by a significant reduction in the Na+ excretion was observed six hours after Pb exposure; this effect persisted for 24 hours. TBARS levels increased in kidney cortex 24 hours after Pb administration. In kidney cortex, Pb exposure affected the expression of HO-1, a renal protein associated with oxidative stress. HO-1 mRNA increased 2.3-fold, three hours after Pb administration and remained increased for six, 12 and 24 hours. HO enzymatic activity and HO-1 protein increased six and three hours after Pb administration, respectively, and remained increased at 24 hours. HO inhibition by tin protoporphyrin, potentiated Pb-induced increase in TBARS and prevented the Pb induced reduction in Na+ excretion. Our data suggest that Pb may be acting through the generation of oxidant products and induction of HO. PMID- 12774887 TI - Protective role of DLalpha-lipoic acid against adriamycin-induced cardiac lipid peroxidation. AB - The cytoprotective activity of alpha-lipoic acid against free radical toxicity manifested during adriamycin (ADR)-induced cardiotoxicity has been investigated. ADR is a potent antitumour drug known to cause severe cardiotoxicity. Although ADR generates free radicals, the role of these radicals in the development of cardiac toxicity is still not well understood. In the present study, we evaluated the influence of chronic ADR treatment on the cellular defence mechanism against free radicals and the effect of alpha-lipoic acid supplementation on ADR-induced cardiotoxicity in male Wistar rats. The increase in lipid peroxidation (LPO) and activities of serum myocardial enzymes, namely lactate dehydrogenase (LDH) and creatinephosphokinase, associated with the decrease in activities of enzymatic (SOD, CAT, GPx, G6PD and GR) and non-enzymatic (GSH, Vit C and Vit E) antioxidants levels were the salient features observed in ADR-induced cardiotoxicity. Lipoic acid pretreated groups showed significant increase in activities of both enzymatic and non-enzymatic antioxidant levels. These observations highlight the antioxidant property of alpha-lipoic acid and its cytoprotective action against ADR-induced cardiotoxicity. PMID- 12774886 TI - Changes in plasma lipids after a non-lethal dose of cycloheximide in rats. AB - This paper describes a study of the effect of a single intraperitoneal non-lethal dose of cycloheximide (CHM; 2.0 mg/kg body weight) on the concentration of plasma lipids and lipoproteins in male rats killed one, two, three, four and nine days after receiving the dose. The concentration of triglycerides, total cholesterol, high-density lipoproteins (HDL)-cholesterol and low-density lipoproteins (LDL) cholesterol was measured in treated and control animals. The effect of CHM on the concentration of triglycerides, total cholesterol, HDL-cholesterol, and LDL cholesterol was visible in rat plasma throughout the study. Total cholesterol and HDL-cholesterol concentrations showed the same pattern of changes, probably due to the reversible inhibition of apolipoprotein apo A-I synthesis by CHM. The concentration of triglycerides decreased after a lag period of three days when the reserves of apolipoprotein apo B, the main apolipoprotein of very low-density lipoproteins (VLDL)-cholesterols produced in the liver, were consumed. PMID- 12774888 TI - Clinical manifestations of VX poisoning following percutaneous exposure in the domestic white pig. AB - Nerve agents are a class of organophosphorus chemicals that inhibit certain cholinesterase enzymes (ChE). If untreated, percutaneous exposure to nerve agents, such as VX (O-ethyl-S-[2(diisopropylamino)ethyl] methylphosphonothioate) can cause paralysis, apnoea and death. Much of the information concerning the percutaneous absorption and subsequent toxicity of nerve agents has been obtained using various rodent models. However, the most relevant 'skin model' is arguably the pig. Therefore, the purpose of this study was to examine the clinical manifestations of VX intoxication in the domestic white pig following a 2 LD50 (120 microg/kg) percutaneous challenge. There was a consistent onset of signs (where present) in each animal: mastication was followed by miosis, salivation, fasciculations and apnoea. Whilst ChE activity did not correlate with the onset of signs, there was a qualitative relationship in that mastication preceded substantial ChE inhibition, miosis lagged behind the linear decrease in acetylcholinesterase (AChE) activity and fasciculations and apnoea occurred after maximum ChE inhibition had been attained (5-10% of normal). These observations may be of use for the triage of patients exposed to VX. In comparison with similar studies with GD, VX did not affect glucose utilization. However, VX was similar to GD in that it caused a mild hyperkalaemia and hyperphosphataemia, although the significance of this observation was not clear. There was substantial lateral diffusion of the initial droplet of VX over the application site, indicating that, when decontaminating exposed skin, attention should also be directed to areas peripheral to the original site of exposure. PMID- 12774889 TI - Differential responses of rat alveolar macrophages to carpet dust in vitro. AB - Epidemiological studies of workers in carpet weaving units in carpet industries have shown a direct relation between the concentration of carpet dust in the air and respiratory symptoms. To predict the health risk of carpet weavers, this preliminary study was conducted to evaluate the toxic potential of different types of workplace dust by using alveolar macrophages (AMs). Several parameters were observed for cytotoxicity such as cell viability, the release of lactate dehydrogenase (LDH) in rat AMs treated with different concentration of carpet dust and haemolytic potential of erythrocytes. In addition, reactive oxygen/nitrogen species-inducing effects of carpet dust were assessed by nitric oxide (NO), reduced glutathione (GSH) release and hydrogen peroxide (H2O2) generation in AMs. Results of cell viability and hemolytic assay showed a direct correlation between increasing the dust concentration with enhancing the toxic effect. Knotted and tufted carpet dust increases the release of LDH, NO, GSH and H2O2 production with increasing dust concentration. Present observations have revealed that dusts collected from tufted carpet weaving units exhibited more toxicity to AMs than knotted carpet dust. These data further suggest that injurious effects of carpet dust to AMs could pave a way to evaluate the toxic potential of the different types of workplace dusts and component(s) involved in it. PMID- 12774890 TI - The role of the cytoskeleton in mechanotransduction in human osteoblast-like cells. AB - The regulation of osteoblast proliferation is a key factor in maintaining bone mass. The enhancement of this process can be achieved by stimulating the proliferation of these cells. Mechanical stimulation is one of the important enhancing factors, but the exact cellular mechanisms of mechanical stimulation, i.e., mechanotransduction, are unknown. In order to investigate the role of the cytoskeleton components in mechanotransduction for cell proliferation, I compared the total DNA content in cultured replicates of osteoblast-like cells derived from three human donors following their exposure to enhancing mechanical stimulation, with and without added specific microtubular and microfilament polymerization blockers (Colchicin and Cytochalasin D, respectively). The results revealed the essential and unique role of the microtubular component of the cytoskeleton in mechanotransduction for proliferation by showing that Colchicin blocked the expected increase in the DNA content after mechanical stimulation of the cultured replicates without altering the total DNA content in replicates at static conditions. Conversely, a specific blockage of the microfilament polymerization presented uniform cytotoxic effect in both static and biomechanically active environments. Since previous reports indicated the essential role of microfilament polymerization for the osteoblast metabolic activity, the results of this study further support the hypothesis that the mechanotransduction mechanisms for proliferation and metabolic activity are mediated by different intracellular pathways. PMID- 12774891 TI - Spermatogenesis recovery in the mouse after iron injury. AB - Alloys used as prosthetic devices for bone/joint replacement include some heavy metals such as chromium, iron, nickel, or titanium. Unfortunately, due to the aggressive nature of the physiological environment, corrosion of these alloys promotes the release of metal ions into the surrounding tissues causing systemic toxic effects. Our previous preliminary studies have demonstrated that iron induced several morphological changes within mice seminiferous epithelium. The aim of the present work was to investigate, over a one-month period, the possibility of recovery of mice seminiferous epithelium, previously damaged by iron. Male Charles River mice were dosed subcutaneously with 0.5 mL of an iron suspension of 538 mg/L +/- 10(-10) mg/L (n = 5) every 72 hours during two weeks, followed by a recovery period of 30 days. Fragments of the seminiferous tubules were fixed in glutaraldehyde and prepared for light and transmission electron microscopy. Regeneration of spermatogenesis was noted after a one-month period, as illustrated by the presence of normal germ cells, in the usual position within the seminiferous tubules. These germinal elements and the Sertoli cells have shown normal cytological features. These results strongly suggest that the deleterious effects induced by iron are reversible. The presence of residual bodies within Sertoli cells cytoplasm indicates that they are able to perform a normal functional activity in a recovered spermatogenesis. PMID- 12774892 TI - Interaction of alcohol and drugs in fatal poisonings. AB - In Finnish data from 1995-2000, 1006 fatal poisonings due to alcohol (ethanol), a single drug or both were statistically analysed in retrospect to evaluate the interaction between alcohol and drugs. In 53% of these cases, low concentrations of some common benzodiazepines were present. The median postmortem blood alcohol concentration (BAC) was 3.3 percent per thousand (w/w) in the 615 alcohol poisonings, but significantly lower, ranging from 1.3 to 1.7 percent per thousand, when promazine, doxepin, amitriptyline or propoxyphene were found together with alcohol. When levomepromazine, temazepam or zopiclone were present, the median BAC was also significantly lower, 2.5-2.7 percent per thousand. Citalopram and diltiazem did not exhibit a significant effect. The median BAC was significantly lower in cases with high concentrations than in those with low concentrations of a drug (excluding citalopram), suggesting a positive concentration-effect relationship. Fatal toxicity indices (FTIs) were calculated by relating the number of deaths caused by a drug to the corresponding sales figures. Promazine had an extremely high FTI, followed by levomepromazine, propoxyphene, doxepin and amitriptyline. The other drugs had relatively low FTIs. The results reflect not only the acute toxicity of a given drug-alcohol combination but also the manners of use and abuse of these drugs. PMID- 12774893 TI - Localization and segmentation of aortic endografts using marker detection. AB - A method for localization and segmentation of bifurcated aortic endografts in computed tomographic angiography (CTA) images is presented. The graft position is determined by detecting radiopaque markers sewn on the outside of the graft. The user indicates the first and the last marker, whereupon the remaining markers are automatically detected. This is achieved by first detecting marker-like structures through second-order scaled derivative analysis, which is combined with prior knowledge of graft shape and marker configuration. The identified marker centers approximate the graft sides and, derived from these, the central axis. The graft boundary is determined by maximizing the local gradient in the radial direction along a deformable contour passing through both sides. Three segmentation methods were tested. The first performs graft contour detection in the initial CT-slices, the second in slices that were reformatted to be orthogonal to the approximated graft axis, and the third uses the segmentation from the second method to find a more reliable approximation of the axis and subsequently performs contour detection. The methods have been applied to ten CTA images and the results were compared to manual marker indication by one observer and region growing aided segmentation by three observers. Out of a total of 266 markers, 262 were detected. Adequate approximations of the graft sides were obtained in all cases. The best segmentation results were obtained using a second iteration orthogonal to the axis determined from the first segmentation, yielding an average relative volume of overlap with the expert segmentations of 92%, while the interexpert reproducibility is 95%. The averaged difference in volume measured by the automated method and by the experts equals the difference among the experts: 3.5%. PMID- 12774894 TI - Construction of an abdominal probabilistic atlas and its application in segmentation. AB - There have been significant efforts to build a probabilistic atlas of the brain and to use it for many common applications, such as segmentation and registration. Though the work related to brain atlases can be applied to nonbrain organs, less attention has been paid to actually building an atlas for organs other than the brain. Motivated by the automatic identification of normal organs for applications in radiation therapy treatment planning, we present a method to construct a probabilistic atlas of an abdomen consisting of four organs (i.e., liver, kidneys, and spinal cord). Using 32 noncontrast abdominal computed tomography (CT) scans, 31 were mapped onto one individual scan using thin plate spline as the warping transform and mutual information (MI) as the similarity measure. Except for an initial coarse placement of four control points by the operators, the MI-based registration was automatic. Additionally, the four organs in each of the 32 CT data sets were manually segmented. The manual segmentations were warped onto the "standard" patient space using the same transform computed from their gray scale CT data set and a probabilistic atlas was calculated. Then, the atlas was used to aid the segmentation of low-contrast organs in an additional 20 CT data sets not included in the atlas. By incorporating the atlas information into the Bayesian framework, segmentation results clearly showed improvements over a standard unsupervised segmentation method. PMID- 12774895 TI - Three-dimensional motion tracking of coronary arteries in biplane cineangiograms. AB - A three-dimensional (3-D) method for tracking the coronary arteries through a temporal sequence of biplane X-ray angiography images is presented. A 3-D centerline model of the coronary vasculature is reconstructed from a biplane image pair at one time frame, and its motion is tracked using a coarse-to-fine hierarchy of motion models. Three-dimensional constraints on the length of the arteries and on the spatial regularity of the motion field are used to overcome limitations of classical two-dimensional vessel tracking methods, such as tracking vessels through projective occlusions. This algorithm was clinically validated in five patients by tracking the motion of the left coronary tree over one cardiac cycle. The root mean square reprojection errors were found to be submillimeter in 93% (54/58) of the image pairs. The performance of the tracking algorithm was quantified in three dimensions using a deforming vascular phantom. RMS 3-D distance errors were computed between centerline models tracked in the X ray images and gold-standard centerline models of the phantom generated from a gated 3-D magnetic resonance image acquisition. The mean error was 0.69 (+/- 0.06) mm over eight temporal phases and four different biplane orientations. PMID- 12774896 TI - Mixtures of general linear models for functional neuroimaging. AB - We set out a new general framework for making inferences from neuroimaging data, which includes a standard approach to neuroimaging analysis, statistical parametric mapping (SPM), as a special case. The model offers numerous conceptual and statistical advantages that derive from analyzing data at the "cluster level" rather than the "voxel level" and from explicit modeling of the shape and position of clusters of activation. This provides a natural and principled way to pool data from nearby voxels for parameter and variance-component estimation. The model can also be viewed as performing a spatio-temporal cluster analysis. The parameters of the model are estimated using an expectation maximization (EM) algorithm. PMID- 12774897 TI - Slice-to-volume registration and its potential application to interventional MRI guided radio-frequency thermal ablation of prostate cancer. AB - In this study, we registered live-time interventional magnetic resonance imaging (iMRI) slices with a previously obtained high-resolution MRI volume that in turn can be registered with a variety of functional images, e.g., PET, SPECT, for tumor targeting. We created and evaluated a slice-to-volume (SV) registration algorithm with special features for its potential use in iMRI-guided radio frequency (RF) thermal ablation of prostate cancer. The algorithm features included a multiresolution approach, two similarity measures, and automatic restarting to avoid local minima. Imaging experiments were performed on volunteers using a conventional 1.5-T MR scanner and a clinical 0.2-T C-arm iMRI system under realistic conditions. Both high-resolution MR volumes and actual iMRI image slices were acquired from the same volunteers. Actual and simulated iMRI images were used to test the dependence of SV registration on image noise, receive coil inhomogeneity, and RF needle artifacts. To quantitatively assess registration, we calculated the mean voxel displacement over a volume of interest between SV registration and volume-to-volume registration, which was previously shown to be quite accurate. More than 800 registration experiments were performed. For transverse image slices covering the prostate, the SV registration algorithm was 100% successful with an error of <2 mm, and the average and standard deviation was only 0.4 mm +/- 0.2 mm. Visualizations such as combined sector display and contour overlay showed excellent registration of the prostate and other organs throughout the pelvis. Error was greater when an image slice was obtained at other orientations and positions, mostly because of inconsistent image content such as that from variable rectal and bladder filling. These preliminary experiments indicate that MR SV registration is sufficiently accurate to aid image-guided therapy. PMID- 12774898 TI - Imaging of human tooth enamel using ultrasound. AB - This paper reports the results of a complete circumferential scan of a human tooth and its underlying dentino-enamel junction using ultrasound at frequencies in the 10-MHz range. The imagery shows clearly a two-dimensional contour of the dentinoenamel junction with a depth and lateral resolution of approximately 100 microm and 750 microm, respectively. The resulting sonograph is compared with an optical micrograph of the same tooth to verify the accuracy of the ultrasonic technique. The results are a significant step toward the biolocation of submillimeter size features within the tooth volume. PMID- 12774899 TI - A new clutter rejection algorithm for Doppler ultrasound. AB - Several strategies, known as clutter or wall Doppler filtering, were proposed to remove the strong echoes produced by stationary or slow moving tissue structures from the Doppler blood flow signal. In this study, the matching pursuit (MP) method is proposed to remove clutter components. The MP method decomposes the Doppler signal into wavelet atoms that are selected in a decreasing energy order. Thus, the high-energy clutter components are extracted first. In the present study, the pulsatile Doppler signal s(n) was simulated by a sum of random-phase sinusoids. Two types of high-amplitude clutter signals were then superimposed on s(n): time-varying low-frequency components, covering systole and early diastole, and short transient clutter signals, distributed within the whole cardiac cycle. The Doppler signals were modeled with the MP method and the most dominant atoms were subtracted from the time-domain signal s(n) until the signal-to-clutter (S/C) ratio reached a maximum. For the low-frequency clutter signal, the improvement in S/C ratio was 19.0 +/- 0.6 dB, and 72.0 +/- 4.5 atoms were required to reach this performance. For the transient clutter signal, ten atoms were required and the maximum improvement in S/C ratio was 5.5 +/- 0.5 dB. The performance of the MP method was also tested on real data recorded over the common carotid artery of a normal subject. Removing 15 atoms significantly improved the appearance of the Doppler sonogram contaminated with low-frequency clutter. Many more atoms (over 200) were required to remove transient clutter components. These results suggest the possibility of using this signal processing approach to implement clutter rejection filters on ultrasound commercial instruments. PMID- 12774900 TI - Segmentation of prostate boundaries from ultrasound images using statistical shape model. AB - This paper presents a statistical shape model for the automatic prostate segmentation in transrectal ultrasound images. A Gabor filter bank is first used to characterize the prostate boundaries in ultrasound images in both multiple scales and multiple orientations. The Gabor features are further reconstructed to be invariant to the rotation of the ultrasound probe and incorporated in the prostate model as image attributes for guiding the deformable segmentation. A hierarchical deformation strategy is then employed, in which the model adaptively focuses on the similarity of different Gabor features at different deformation stages using a multiresolution technique, i.e., coarse features first and fine features later. A number of successful experiments validate the algorithm. PMID- 12774901 TI - Multielement synthetic transmit aperture imaging using temporal encoding. AB - A new method to increase the signal-to-noise ratio (SNR) of synthetic transmit aperture imaging is investigated. The approach utilizes multiple elements to emulate a spherical wave, and the conventional short excitation pulse is replaced by a linear frequency-modulated (FM) signal. The approach is evaluated in terms of image quality parameters in comparison to linear array imaging. Field II simulations using an 8.5-MHz linear array transducer with 128 elements show an improvement in lateral resolution of up to 30% and up to 10.75% improvement in contrast resolution for the new approach. Measurements are performed using our experimental multichannel ultrasound scanning system, RASMUS. The designed linear FM signal obtains temporal sidelobes below -55 dB, and SNR investigations show improvements of 4-12 dB. A 30 mm (approximately 45%) increase in penetration depth is obtained on a multitarget phantom with 0.5 dB/[cm MHz] attenuation. Furthermore, in vivo images of the abdomen are presented, which demonstrate the clinical application of the new approach. PMID- 12774902 TI - Estimating coronary artery lumen area with optimization-based contour detection. AB - A modified optimization-based contour detection method was presented to compute the lumen area of the coronary artery from intravascular ultrasound (IVUS) video images. First, the search range for the artery inner wall was determined based on the continuity of IVUS video frames. Next, the internal and external energy were calculated to describe the smoothness of the arterial wall and the grayscale variation of ultrasound images, respectively. Here, a novel form of the external energy which combines the gradient and variance of the intensity of image in the radial direction was used. Finally, the minimal energy path based on the optimum contour of the artery wall was obtained using circular dynamic programming (DP). By the comparison with the typical DP procedure using the traditional external energy form, based only on the image gradient, the reliability of this modified method is considerably improved in the measurement of coronary artery lumen area. PMID- 12774903 TI - Removal of organic pollutants and metabolic adaptation of microorganisms by micro aeration. AB - Aerobic systems are high rate processes for wastewater treatment, but they consume much power for aeration. Several low-cost anaerobic systems have been developed for mid- and low-income countries. The removal efficiencies of nutrients, particulate matter, and pathogens are not satisfactory for the anaerobic systems. Micro-aeration is a type of low-cost aerobic application which enhances oxygen transfer through water surface. Trickling was found the most efficient method of micro-aeration and batch experiments were performed to evaluate the effectiveness. When soluble organic carbon was removed, ORP7 (redox potential rectified to pH at 7) were around -120 or +260 mV. When soluble organic carbon increased due to organic overload, ORP7 was around +30 mV. At +30 mV, carbon fraction of biomass increased to 0.54 (higher than average value of 0.46). It was presumable that the ORP7 of -120, +260, and +30 mV were anaerobic, aerobic, and internal storage mode, respectively. Maximum reaction rate of aerobic condition was four times as high as that of anaerobic condition. At internal storage mode, the rate was halved from aerobic mode. It is concluded that microorganisms adapt the metabolic systems to conform to the redox environment, which can be evaluated by ORP. Organic overload results metabolic shift to internal storage mode which retards mineralization of organic matter. PMID- 12774904 TI - Nitrogen removal via nitrite at normal temperature in A/O process. AB - In order to study the nitrogen removal via nitrite at normal temperature in anoxic/oxic (A/O) process for treating domestic wastewater, the influence of pH, free ammonia (FA), dissolved oxygen (DO), and hydraulic retention time (HRT) were studied. Results indicated that it was possible to remove nitrogen via nitrite in A/O process, if the temperature was between 18 and 25 degrees C and pH was below 7.5. Even if FA was as low as 0.06 mg NH3-N/L, it would inhibit the nitrobacteria. However, FA could not be the sole factor. Whether denitrification went thoroughly or not could affect the nitrification pathway. If denitrification went well, nitrite accumulation could recover in a short while. Nitritification lagged nitratification, so that short HRT would help nitrite accumulation. On the other hand, extended aeration would reduce nitrite accumulation. PMID- 12774905 TI - Occupational exposure to urban pollutants and plasma growth hormone (GH). AB - The aim of present study was to evaluate whether traffic policemen exposed to urban pollutants and possible psycho-social stressors are at risk of alterations on plasma growth hormone (GH) levels compared with a control group. Out of a population of 395 Municipal Police employees, the subjects with principal confounding factors were excluded from the study. The remaining traffic policemen were matched with those not exposed by sex, age, working life, and habitual consumption of alcohol; 71 traffic policemen (40 men and 31 women) with outdoor activity exposed to urban pollutants and 71 not exposed subjects (40 men and 31 women) with indoor activity were included in the study. The plasma levels of GH were significantly lower in the exposed traffic policemen compared with those not exposed (P = 0.000); similarly in male (P = 0.011) and female subjects (P = 0.000). The authors hypothesize the possibility of an effect of the specific working activity in traffic policemen on the plasma GH concentrations. PMID- 12774906 TI - Quantitative prediction of traffic pollutant transmission into buildings. AB - An integrated air quality model that combines a CFD model and multi-room pollutant transport model has been developed to study the effect of traffic pollution on indoor air quality of a multi-room building located in close proximity to busy roads. The CFD model conducts the large eddy simulation of the three-dimensional turbulent flows and pollutant transport processes in outdoor, whereas the multi-room pollutant transport model performs zonal airflow and pollutant transport in indoor. The integrated model is verified with available field measurement of traffic-induced CO concentrations. Twelve scenarios of numerical experiments for various configurations of window openness are carried out to study the effects of the air change rate and the outdoor pollutant dispersion on indoor air quality. It is concluded that the windward side opening is a significant factor contributing to indoor air quality. Using air inlets on the sideward and leeward envelopes simultaneously can effectively lower the daily mean and peak indoor levels of traffic pollutants and maintain a desirable air change rate. PMID- 12774907 TI - Application of intake fraction to population exposure estimates in Hunan Province of China. AB - This article developed a new method, based on the concept of "intake fraction", to assess population exposure to primary and secondary fine particle matters from site-specific sources. This method was illustrated by a set of 17 power plants (totally 24 stacks) in Hunan Province of China. The CALPUFF long-range atmospheric dispersion model was used to simulate ambient concentrations of fine particles, and the GIS technology was used to generate a population distribution database from county-level population statistical data. An integrated computer program package was developed to carry out numerical integration of dispersion results over the population data, and produce intake fractions. The resulting average intake fractions within 500 km were 9.73 x 10(-6), 2.39 x 10(-6), and 2.47 x 10(-6) for primary fine particles, sulfate and nitrate respectively. Regression analyses were performed to explore the relationships between intake fractions and potential variables. Results showed that the stack height and aggregate populations could be used to predict intake fractions of fine particles. R2 of the regression equations were 0.83, 0.64, and 0.74 for primary fine particles, sulfate and nitrate respectively. Iso-intake fractions presenting geographical distributions of intake fractions in Hunan Province were mapped, showing a factor of about 2 between the highest values in the northeast and the lowest in the southwest of Hunan Province. PMID- 12774908 TI - Oxidative degradation of indigocarmine by hypochlorite--a tool for determination of hypochlorite in commercial samples. AB - The oxidative action of hypochlorite on indigocarmine is investigated. Hypochlorite dissociates indigocarmine to produce isatin-5-monosulphonic acid, with a stoichiometry of 2:1. The probable mechanism is proposed. The suitability of reaction between hypochlorite and indigocarmine as an indicator reaction for the determination of high levels of hypochlorite in synthetic and commercial samples is investigated. A simple and reliable kinetic-analytical method is developed for the determination of hypochlorite ion at alkaline pH, based on its oxidizing activity. PMID- 12774909 TI - Developmental and reproductive effects of low cadmium concentration on Artemia parthenogenetica. AB - Hatching, reproductive, and lifespan characteristics of an Artemia parthenogenetica population from La Mata (Alicante, Spain) exposed to cadmium were studied. No effects on percentage of cyst hatched nor time of hatching were observed on cysts exposed from 0.01 to 5 mg Cd/L. Cohorts of Artemia were chronically exposed to cadmium in life-table experiments to test whether ecologically relevant impacts may occur after several generations of exposure to a very low concentration of cadmium. Chronic toxicity tests were performed using neonates of a third generation of shrimps exposed to 0.08 mg Cd/L (1/1000 of the LOEC for 24 h acute toxicity test). No significant reduction in the intrinsic rate of natural increase (rm) was found as a result of cadmium exposure, although the age-specific fecundities of the cadmium exposed shrimps were lower than those of the controls. A significant decrease in the prereproductive period and a reduction in the percentage of fertile females was also found as a consequence of the metal exposure. PMID- 12774910 TI - Removal of total suspended solids from wastewater in constructed horizontal flow subsurface wetlands. AB - Subsurface horizontal flow experimental wetlands (reed beds), were designed and built based on a combination of two design methodologies, that of the WRc and Severn Trent Water plc (1996) and that of the USA, EPA (1988). Four different growing media were used with a combination of top soil, gravel, river sand, and mature sewage sludge compost, to determine the best substrate for total suspended solids (TSS) removal. Eight units were constructed, two for each growing media. One bed for each pair was planted with Typha latifolia plants commonly known as cattails. Primary treated domestic wastewater, was continuously fed to the beds for more than six months. All eight beds performed very well. The best performance was achieved by the gravel reed beds with an almost constant removal rate above 95% and an average effluent concentration of less than 10 mg/L. Soil based beds containing top soil and sand, managed to reach values of removal around 90%. The wetlands containing compost in their substrate, produced an effluent with average concentration of less than 30 mg/L and a percentage removal between 80% and 90%. As expected, there was no significant difference in the performance of planted and unplanted wetlands. PMID- 12774911 TI - Anaerobic biodegradation of toluene coupled to sulfate reduction in oil contaminated soils: optimum environmental conditions for field applications. AB - Degradation characteristics of toluene in enrichment culture were investigated in soil microcosms study and the optimum environmental conditions for anaerobic degradation of toluene coupled with sulfate reduction were determined for field site applications. Anaerobic sulfate-reducing bacteria were enriched from oil contaminated soil samples with toluene. Enriched consortia degraded toluene with sulfate as a terminal electron acceptor. The average degradation rate of toluene in enriched consortia ranged from 0.08 to 0.1 micromol g(-1) day(-1). Toluene degradation under sulfate-reducing condition was inhibited in the presence of molybdate alone or together with nitrate or fumarate, indicating that toluene is degraded directly by sulfate-reducing bacteria. Effects of initial toluene concentration, pH, temperature, and other hydrocarbons on toluene degradation were investigated. There is a trend of increasing rate of toluene degradation with increasing the initial mass up to 94 micromol of toluene. Toluene degradation did not affected by the presence of ethylbenzene and xylene, as a while, toluene was degraded in a slower rate in the presence of benzene. The sulfate-reducing bacteria in the enriched culture showed higher microbial activity at neutral (pH 6-8) and medium temperature (30-37 degrees C) environments. PMID- 12774913 TI - An assessment of aquatic ecosystem health in a temperate watershed using the index of biological integrity. AB - The health effect of an aquatic ecosystem on habitat modifications were evaluated in the Keum river watershed, Korea during 1977-1996 using the Index of Biological Integrity (IBI) based on fish assemblages. Values of IBI, based on overall sites, averaged 35 (range: 26-45, n = 38) before dam construction, indicating a "fair health condition" based on the modified criteria of Karr and Chu (Karr, J.R.; Chu, E.W. Restoring Life in Running Waters: Better Biological Monitoring; Inland Press: Washington, DC, 1999; 206 pp.), while the values averaged 33 (range: 18 48, n = 15) after dam construction, indicating a similar ecosystem health condition in the IBI between the two periods. Marked modifications in the IBI, however, were partially observed along the longitudinal gradients from the headwaters to downstream along with variations of trophic compositions and habitat guilds. Annual mean of IBI showed significant decreases (p < 0.001, t = 10.03) in the mid-reach of 100-240 km location after the construction along with >20% decreases of insectivores and >25% increases of omnivores. Comparisons of habitat guilds indicated that the proportion of riffle benthic species declined linearly from 1977 to 1996 and had inverse relations (r = -0.78, p < 0.01) with that of water column species. Such variations were explained by serial discontinuity concept that was developed by Ward and Stanford (Ward, J.V.; Stanford, J.A. The serial discontinuity concept of lotic ecosystems. In Dynamics of Lotic Ecosystems; Fontaine, J.V., Bartell, S.M., Eds.; Ann Arbor Science: Ann Arbor, Michigan, USA, 1983; 29-42). Chemical data of long-term BOD5 and COD5 indicated that chemical impacts after the dam construction were minor compared to the condition before the construction. Overall variation of IBI was highly accounted (p < 0.001, R2 = 0.91, n = 38) by the Qualitative Habitat Evaluation Index (QHEI), suggesting that the ecosystem health was mainly affected by the habitat modifications. PMID- 12774912 TI - Effects of the headspace gas composition on anaerobic biotransformation of o-, m , and p-toluic acid in sediment slurries. AB - Composition of the headspace gas affected the biotransformation pattern of toluic acid isomers in anoxic sediment slurries. Under an N2 atmosphere, o- and m-, and p-toluic acid (20-25 mg L(-1)) were biotransformed in 100 days, 77 days, and 148 days, respectively, with a lag period of 50 days, 49 days, and 50 days, respectively. Under a CO2 atmosphere, the same toluic acid isomers were biotransformed by the sediment microorganisms in 16-25 days without a lag period. CO2 thus increased the biotransformation rates. The presence of H2, on the other hand, decreased the biotransformation rates: in most cases, adding H2 gas (5% and 20% to the N2 and CO2 atmospheres, respectively) not only increased the lag period but also decreased the maximum biotransformation rates. These effects were especially noticeable for the N2 atmosphere. Under N2, the maximum biotransformation rates of the toluic acid isomers were in the order o-toluic acid > m-toluic acid > p-toluic acid. However, under CO2, the maximum biotransformation rates were reversed, i.e., p-toluic acid > m-toluic acid > o toluic acid. The presence of the methanogen inhibitor bromoethanesulfonic acid (BESA) slowed the biotransformation rates of p-toluic acid, and this together with the population dynamics of the acetogenic bacteria in the sediment slurries, suggested that acetogenic bacteria were involved in the degradation pathway. However, their exact role remains unclear. PMID- 12774914 TI - Heterogeneous photocatalytic oxidation of acetone for airpurification by near UV irradiated titanium dioxide. AB - This work presents a photocatalysis-based method to treat and purify air because of its broad applicability to common, oxidizable air contaminants. The effect of oxygen content, temperature, water vapor, and acetone concentration on the photooxidation of acetone on TiO2 surface was investigated. The photocatalytic decomposition reaction of acetone obeyed the first-order equation. The decomposition rate increased with increasing the oxygen content. The rate of acetone oxidation increased when water vapor increased from 18.7 to 417 microM and decreased at higher than 417 microM. The conversion and mineralization of acetone decreased at higher than 138 degrees C. The initial rate of acetone degradation can be well described by the Langmuir-Hinshelwood rate form. The specific reaction rate constant and the equilibrium adsorption are 15.8 microM/min and 0.0671 L/microM, respectively. The difference between observed and estimated half-lives became larger when the initial concentration of acetone was increased. It is assumed that the intermediates competed with parent compound so that delayed the half-life. The detection of CO2 production can support this assumption. PMID- 12774915 TI - Sorption and desorption kinetics of surfactants TX-100 and DPC on different fractions of soils. AB - Surfactant-based technologies are promising remediation alternatives. The information on sorption and desorption kinetics of surfactants on soils is important in the successful application of surfactant-based technologies. In this study, the sorption and desorption rates of nonionic surfactant TX-100 and cationic DPC were correlated to the surfactant concentration, soil organic matters (SOM), and soil cation exchange capacity (CEC). The results indicated that at higher initial surfactant concentrations, sorption rates of surfactants increased linearly with SOM and soil CEC for TX-100 and DPC, respectively. The sorption rates and initial surfactant concentrations followed the first order relation for TX-100 and second order for DPC. A linear relationship between the sorption rates of surfactants and soil characteristics was developed. The desorption rates of TX-100 and DPC increased linearly with the increased surfactant levels sorbed on soils but were irrelevant to soil characteristics and the contact time of surfactant sorption. The rate of surfactant desorption was similar as the amount of surfactants sorbed on soils was in the same range. The cationic DPC sorbed and desorbed at two orders of magnitude faster than the nonionic TX-100, suggesting that both sorption and desorption have to be considered in the remediation process. PMID- 12774916 TI - The catalytic incineration of trichloroethylene over a gamma-alumina supported manganese oxide catalyst. AB - TCE decomposition over a Mn2O3/gamma-Al2O3 catalyst in a fixed bed reactor was conducted in this study. Preliminarily, three catalysts including Mn2O3/gamma Al2O3, NiO/gamma-Al2O3, and Pt/gamma-Al2O3 were used to incinerate TCE and the results show that the Mn2O3/gamma-Al2O3 catalyst has the best performance. The effects of operating parameters, such as inlet temperature, space velocity, TCE inlet concentration, and oxygen concentration on the catalytic incineration of TCE over the Mn2O3/gamma-Al2O3 catalyst were then performed. The results show that conversion of TCE increases as inlet temperature and oxygen concentration increase, and decreases with the increases of TCE concentration and space velocity. The activity of the catalyst decreases significantly while TCE incineration is operated under a low temperature, 365 degrees C. However, the activity of the catalyst does not change much while the operating temperature is as high as 500 degrees C. The catalysts were characterized by the surface and pore size analysis, XRD, XPS, EDS, and SEM before and after the tests. The results show that the catalytic crystal is Mn2O3, the catalytic deactivation is not due to carbonaceous material, and the chlorine element is adsorbed on the surface of catalysts. PMID- 12774917 TI - The pathogenetic role of oncogenes deregulated by chromosomal translocation in B cell malignancies. AB - Chromosomal translocations involving the immunoglobulin (IG) loci play a pivotal role in the pathogenesis of many subtypes of mature B-cell malignancy. Although all the common IG translocations have been cloned, cloning of rare but nonetheless recurrent translocations continues to allow identification of genes of importance to the development of both normal and malignant B-cells. Clustering of breakpoints within the IG gene segments has allowed development of polymerase chain reaction methods that facilitate cloning. IG translocations result in overexpression of a wide variety of genes ranging from cell surface receptors to transcriptional repressors. Genes recently shown to be involved in such translocations include BCL11A and MALT1. As with the acute leukemias, different translocations in B-cell lymphomas may target different proteins that interact directly. A common endpoint for several translocations is activation of the nuclear factor kappaB pathway. Analysis of the mechanisms of transformation may define new therapeutic strategies. PMID- 12774918 TI - Diffuse large B-cell lymphoma: insights gained from gene expression profiling. AB - Analysis of global gene expression with DNA microarrays has great potential to improve the understanding of tumorigenesis advance tumor diagnosis and classification, and affect cancer treatment. Diffuse large B-cell lymphoma (DLBCL) is the most common type of non-Hodgkin's lymphoma. However, we now realize that the disease is extremely heterogeneous. This review summarizes the progress in understanding DLBCL that has been made as a result of the application of gene expression profiling. PMID- 12774919 TI - The molecular basis for the generation of Hodgkin and Reed-Sternberg cells in Hodgkin's lymphoma. AB - Hodgkin's lymphoma (HL) is a lymphoid neoplasm with a low frequency of malignant tumor cells, known as Hodgkin and Reed-Sternberg (H-RS) cells, in a background of mixed cellular infiltrates. Despite extensive studies on H-RS cells, the molecular mechanisms of their growth and regulation have remained uncertain for a long period. Recently, constitutively activated nuclear factor-kappaB (NF-kappaB) was reported to be a unique and common characteristic of H-RS cells that prevents the cells from undergoing apoptosis. NF-kappaB triggers proliferation and provides a molecular basis for these cells' aberrant growth and cytokine gene expression. In HL pathogenesis associated with Epstein-Barr virus infection, the activation of NF-kappaB is induced by viral latent membrane protein 1 (LMP1). Coupled with recent insights into the molecular mechanisms of activation of NF kappaB signaling in H-RS cells, this review discusses a linkage between LMP1 and HL via CD99, which has recently been reported to be down-regulated by LMP1 through the NF-kappaB signaling pathway. This down-regulation leads to the generation of cells with H-RS phenotypes related to the clinical and histologic characteristics of HL. PMID- 12774920 TI - Immunodeficient mouse models of lymphoid tumors. AB - Severe combined immunodeficient (SCID) mice lack functional T- and B-cells and readily accept human xenografts, including hematopoietic malignancies. Accordingly, SCID mice have been used to study the growth and behavior of lymphoid tumors in vivo. The SCID mouse models of disease mimic human diseases and have provided valuable information. However, this mouse strain has some residual immunity that somewhat limits posttransplantation growth of human xenografts. Recently, the SCID mutation was backcrossed onto the nonobese diabetic (NOD) strain background. The result was an animal with additional immunological defects beyond those seen in SCID mice. The NOD/SCID strain appears to be more promising as a tool for xenotransplantation of lymphoid tumors. Moreover, these SCID and NOD/SCID mouse models have been used to develop novel therapeutic strategies. Results from such studies may also help to elucidate the pathogenesis of lymphoid tumors. PMID- 12774922 TI - Relationship between temperature and apoptosis of human peripheral blood mononuclear cells. AB - To examine the effect of various incubation temperatures on the apoptotic death of human peripheral blood mononuclear cells (PBMC), we incubated cells at 37 degrees C, 22 degrees C, and 4 degrees C for 1 and 24 hours. In addition, cells incubated at 4 degrees C for 3, 6, and 9 hours were rewarmed to 37 degrees C until a total incubation time of 24 hours was reached. The percentage of apoptotic cells was detected by a flow cytometric assay using propidium iodide staining. Incubation of PBMC at the above-mentioned temperatures for 1 hour did not affect the percentage of apoptotic cells. However, incubation at 4 degrees C for 24 hours resulted in the lowest percentage of apoptotic cells compared to those incubated at 22 degrees C and 37 degrees C. Rewarming of the cells to 37 degrees C increased the percentage of apoptotic cells to a level similar to that of the controls (incubated at 37 degrees C). Because PBMC are closely involved in the normal function of the immune system, the results of the study should be considered in cases in which these cells are exposed to various thermal conditions. PMID- 12774921 TI - Oxidative stress and the myelodysplastic syndromes. AB - The evolution of higher organisms from anaerobic to aerobic living has promoted an elaborate mechanism of defense against potentially toxic oxidants. Many environmental toxicants implicated in the pathogenesis of myelodysplastic syndromes (MDS), including benzene and ionizing radiation, exert toxicity via pro oxidant mechanisms. The emerging data suggest a probable genetic susceptibility to environmental carcinogenesis through functional polymorphic variants in enzymes that metabolize toxicants and/or protect against oxidative stress. The most studied enzyme is NAD(P)H:quinone oxidoreductase (NQO1). CD34+ cells from individuals homozygous for the NQO1 C609T nonfunctional allelic variant are incapable of enzyme induction following exposure to benzene, thus potentially increasing the hematotoxicity of benzene metabolites. Serologic and molecular markers of oxidative stress are present in many patients with MDS and include an increased concentration of the lipid peroxidation product malondialdehyde and the presence of oxidized bases in CD34+ cells. Potential mechanisms of oxidative stress include mitochondrial dysfunction via iron overload and mitochondrial DNA mutation, systemic inflammation, and bone marrow stromal defects. The biological activity of the antioxidant aminothiol amifostine in vivo suggests that these pathways may be meaningful targets for future therapy in MDS patients. PMID- 12774924 TI - Expression of apoptosis-associated protein RCAS1 in macrophages of histiocytic necrotizing lymphadenitis. AB - Receptor-binding cancer antigen expressed on SiSo cells (RCAS1), which is recognized by the 22-1-1 monoclonal antibody (MoAb) against human uterine adenocarcinoma cell line SiSo, has been identified on various kinds of cancer cells. RCAS1 appears to be an apoptosis-associated protein that induces apoptosis in activated T-cells and erythroid progenitor cells. We previously demonstrated that monocytes/macrophages express RCAS1. In the present study, we investigated RCAS1 expression by 22-1-1 MoAb in histiocytic necrotizing lymphadenitis (HNL), which is characterized by necrotic lesions consisting of T-cells undergoing apoptosis and macrophages in proliferation. Expression of RCAS1 was analyzed by immunohistochemical staining in 9 cases of HNL and in 9 cases of reactive lymphadenitis used as a control. The ratio of RCAS1+ cells to CD68+ cells (monocytes/macrophages) was significantly higher in the patients with HNL than in the patients with reactive lymphadenitis (P = .0002; paired t test). Our findings suggest that RCAS1 expressed on macrophages may play an important role in the induction of activated T-cell apoptosis in cases of HNL. PMID- 12774923 TI - Pure red cell aplasia in a patient with trisomy X chromosome abnormality and reactivated Epstein-Barr virus infection. AB - We describe a woman with a congenital chromosome anomaly, 47,XXX, who developed chronic pure red cell aplasia (PRCA). The patient had serologic reactivity consistent with that of reactivated Epstein-Barr virus (EBV) infection, as judged by high titers for anti-EBV viral capsid antigen (VCA) immunoglobulin G (IgG) and anti-early antigen (EA) IgG. Detection of EBV genome in peripheral blood cells and cell-free serum also supported the diagnosis. Although EBV infection has been implicated in the pathogenesis of acute PRCA, the viral infection rarely results in a chronic disease state. So far, only 1 case of EBV-associated chronic PRCA has been reported, to the best of our knowledge. Chronic PRCA also is known to occur on an autoimmune basis. Individuals carrying an extra X chromosome, such as XXY and XXX, are prone to development of immune abnormalities. Our patient had an anti-DNA autoantibody and a positive result of the direct Coombs test. The pathogenesis of PRCA in this case seemed to involve multiple factors. In addition to the infectious agent, host factors may have played a role. Although the etiologic link between chronic PRCA and trisomy X remains to be elucidated, our findings suggest the importance of karyotype analysis as well as search for infectious agents in patients with chronic PRCA. PMID- 12774926 TI - Size and composition of T-cell receptor delta (TCRD) junctional sequences are not predictive of the sensitivity of clonospecific oligonucleotides designed for detection of minimal residual disease in acute lymphoblastic leukemia. AB - We characterized 168 junctional regions of T-cell receptor delta (TCRD) rearrangements from 116 children with acute lymphoblastic leukemia (ALL) (101 with precursor B-cell ALL, 15 with T-cell ALL). Application of 101 allele specific oligonucleotide (ASO) probes representing 85 Vdelta2Ddelta3, 10 Ddelta2Ddelta3, 3 Vdelta1Jdelta1, 1 Vdelta3Jdelta1, and 2 Ddelta2Jdelta1 junctions for the detection of minimal residual disease (MRD) revealed detection levels of 10(-4) to 10(-6) leukemia cells in the vast majority of cases (93 of 101). Of interest was that neither the N, D, P (nontemplated, diversity, palindromic) content and length of the junctional regions nor the number of nucleotides deleted from the flanking V, D, or J (variable, diversity, joining) elements correlated with the sensitivity of ASO probes. These data indicated that in ALL TCRD rearrangements can serve as suitable tools for the detection of MRD irrespective of the specific composition of the junctional region. PMID- 12774925 TI - Constitutive activation of extracellular signal-regulated kinase and p38 mitogen activated protein kinase in B-cell lymphoproliferative disorders. AB - Signaling molecules such as p21(ras) (Ras), mitogen-activated protein kinase (MAPK), and Akt kinase play pivotal roles in the proliferation and survival of lymphoid cells in response to many kinds of stimulation. It is not fully understood, however, how these molecules participate in the growth of malignant lymphoid cells. We determined whether Ras, MAPKs such as extracellular signal regulated kinase (ERK), c-Jun amino-terminal kinase (JNK), and p38 MAPK, and Akt kinase are activated in B-cell tumors, including acute lymphoblastic leukemia, chronic lymphocytic leukemia, Burkitt-like lymphoma, diffuse large B-cell lymphoma, and plasma cell leukemia. We found that Lyn protein tyrosine kinase was constitutively phosphorylated on tyrosine, and that ERK and p38 MAPK were constitutively active in all cases of the B-cell tumor. In contrast, activation of Ras and Akt kinase was found in limited cases, and JNK kinase activity was not observed in any case. These results suggest that ERK and p38 play roles in the oncogenesis of B-cell tumors. PMID- 12774927 TI - Characterization of blasts in clinical samples containing few blasts. AB - Characterization (eg, phenotyping) of blasts present at low percentages in clinical samples is often required for decisions regarding the approach to therapy. However, the available methods for cell characterization do not yield reliable data when the target cells are scant, and the existing methods for blast enrichment, such as cell sorting by flow cytometry (FCM), cannot enrich blasts of unknown immunophenotype. Blastretriever is a newly developed density centrifugation reagent for retrieving blasts. We examined the utility of Blastretriever in clinical practice. When normal bone marrow (BM) cells were separated with this reagent, myeloblasts and B-cell precursors were enriched and detected as clusters on the FCM cytogram. Compared with a conventional reagent for mononuclear cell preparation, the Blastretriever reagent markedly enriched leukemic myeloblasts, leukemic lymphoblasts, and blastoid lymphoma cells from 36 test samples (BM cells and peripheral blood). We then applied the Blastretriever reagent to samples from 11 consecutive patients who had been referred to us because they exhibited low percentages of blasts (1 patient had only 0.2% blasts). Characterization was needed but impossible with conventional analyses. Blast enrichment was achieved for all 11 samples, allowing reliable blast characterization by FCM, fluorescence in situ hybridization, and/or G-banding determinations. The revealed blast characteristics were valuable for choosing appropriate therapy for the patients. PMID- 12774928 TI - Sweet's syndrome associated with recurrent fever in a patient with trisomy 8 myelodysplastic syndrome. AB - In the case of a 69-year-old man suffering from recurrent high fever, laboratory data demonstrated elevated inflammatory findings such as neutrophilic leukocytosis and an increase in serum level of C-reactive protein. A nonsteroidal antiinflammatory drug or a low dose of prednisolone proved to be temporarily effective. A bone marrow specimen showed myelodysplastic syndrome (MDS) of the refractory anemia (RA) type with trisomy 8. Seven months after onset, painful erythematous eruptions were found on both legs. Biopsy of a skin lesion demonstrated neutrophic infiltration into the dermis. As a result, Sweet's syndrome (SS) was diagnosed on the basis of clinical and histopathological findings. Administration of prednisolone (30 mg/d) was started and resulted in prompt defervescence and resolution of the lesions within 1 week. We also investigated serum levels of 3 cytokines: interleukin 1beta (IL-1beta), IL-6, and granulocyte colony-stimulating factor (G-CSF). Levels of IL-6 and G-CSF were elevated during the active phase, but the level of IL-1beta did not increase. Because cases of MDS with trisomy 8 and SS or Behcet disease have been reported recently, our findings suggested that cytokine production, which enhances neutrophil function, is elevated in some MDS patients with trisomy 8; that is, not only cytokines but also trisomy 8 may be related to the pathogenesis of SS in MDS. It is thus advisable to watch for development of SS during follow-up of MDS patients with recurrent fever or trisomy 8. PMID- 12774929 TI - Non-Hodgkin's lymphoma developing in a pacemaker pocket. AB - A 29-year-old man developed diffuse large B-cell lymphoma in a subpectoral pacemaker pocket that 6 years previously had been created in the chest for a titanium-covered pulse generator. The patient had an 8-cm-diameter dark red tumor with necrotic tissue on a keloidal surgical scar in the left side of the chest. Left axillary lymphadenopathy also was present. Laboratory studies showed an increased level of soluble interleukin 2 receptor and a normal level of lactose dehydrogenase. A biopsy specimen showed a diffuse large B-cell phenotype and monoclonal immunoglobulin H gene rearrangement. A gallium scintigraphy study showed abnormal accumulation in the left chest and left axilla. On the basis of these findings, we diagnosed diffuse large B-cell lymphoma, stage II. The patient received THP-COP chemotherapy (pirarubicin, cyclophosphamide, vincristine, and prednisolone) and radiotherapy, achieved complete remission, and was free of disease for 16 months after treatment. This case suggests that there was a relationship between the development of non-Hodgkin's lymphoma and the presence of chronic inflammation in the pulse generator pocket. PMID- 12774930 TI - Persistent remission after immunosuppressive therapy of hairy cell leukemia mimicking aplastic anemia: two case reports. AB - Some patients with hairy cell leukemia (HCL) manifest pancytopenia and bone marrow hypoplasia without an apparent increase in atypical cells, so their disease resembles severe aplastic anemia at onset. We treated 2 HCL patients, who were initially diagnosed with aplastic anemia, with antithymocyte globulin (ATG) in combination with cyclosporine or antilymphocyte globulin (ALG). Both patients obtained partial remission in response to the immunosuppressive therapy and did not need transfusion treatment for more than 3 years. Sustained improvement of hematopoiesis in such B-cell malignancies after ATG/ ALG therapy suggests that the mechanisms underlying successful immunosuppressive therapy for aplastic anemia may involve B-cell suppression, inhibiting hematopoietic stem cells. PMID- 12774931 TI - Esophageal erosion as a possible bacterial entry site in an acute lymphoblastic leukemia patient with sepsis. AB - A 69-year-old man with relapsed acute lymphoid leukemia was treated with adriamycin, vincristine, and prednisolone. During this chemotherapy, the patient developed sepsis and meningitis. Although many kinds of antimicrobial drugs, including imipenem, meropenem, amphotericin-B, and gamma-globulin were administered, the patient died of respiratory failure. A positive result for Enterococcus faecalis was obtained in both blood and cerebrospinal fluid culture. Autopsy revealed multiple small erosions in the lower esophagus. Histopathological examination showed multiple nuclear inclusion bodies of herpes simplex virus in the squamous epithelial cells at the edge of the erosions. Moreover, proliferation of micrococci was observed at the base of the erosions and in the lumina of the submucosal small vessels. These findings suggested that E faecalis entered the blood circulation from this lesion. In many patients with febrile neutropenia, the pathogenesis of infection remains unclear. Our case seems significant for clarifying the focus and pathogenesis of febrile neutropenia. PMID- 12774933 TI - Rapid improvement of disseminated intravascular coagulation by donor leukocyte infusions in a patient with promyelocytic crisis of chronic myelogenous leukemia after reduced-intensity stem cell transplantation from an HLA 2-antigen mismatched mother. AB - Donor leukocyte infusion (DLI) is recognized as effective therapy for relapse after stem cell transplantation in patients with chronic myelogenous leukemia (CML). However, the clinical efficacy of DLI in the advanced phase of CML or other types of leukemia has not been clearly defined because of its varying degree of success. We describe a 22-year-old male patient with promyelocytic crisis of CML who had a relapse after peripheral blood stem cell transplantation, under reduced-intensity conditioning, from his HLA 2-antigen-mismatched mother. Complete hematologic remission was obtained after transplantation. However, a relapse that occurred on day 66 posttransplantion was characterized by an increase in number of leukemic promyelocytes with simultaneous exacerbation of disseminated intravascular coagulation (DIC). The patient received DLI containing 1 x 10(7)/kg CD3+ cells on day 73. Because rapid improvement of DIC paralleled the decrease in leukemic cells and because it was observed soon after DLI and before the development of acute graft-versus-host disease (GVHD), we hypothesized that leukemia-specific cells other than natural killer cells or cytotoxic T-cells unrelated to GVHD played a role in the graft-versus-leukemia effect observed in our patient. In addition, this may be the first report of effective correction of DIC by DLI after stem cell transplantation. PMID- 12774934 TI - Seroindeterminate HTLV-1 prevalence and characteristics in blood donors in Taiwan. AB - Human T-cell leukemia virus type 1 (HTLV-1) is commonly accepted as the cause of adult T-cell leukemia and tropical spastic paraparesis/HTLV-1-associated myelopathy. Screening of blood donors for HTLV-1 and HTLV-2 was implemented in Taiwan in February 1996. From February 1996 to December 1998, we investigated the seroprevalence of HTLV-1 in all unpaid blood donors in Taiwan. Of 2,578,238 donors in all 6 blood centers, 1793 (0.06%) were seropositive for HTLV-1, and 605 (0.023%) were indeterminate for HTLV-1. Among these indeterminate donors, 359 (59.3%) were male. The most common HTLV-1-indeterminate pattern by Western blot in our study was GD21 alone (34.6%) followed by p24 alone (7.8%), p53 alone (6.5%), and gp46 + GD21 (6.0%). That GD21 pattern was found in 59.6% of indeterminate results in this study suggested that the majority of nonspecific enzyme immunoassay reactions were probably precipitated by viral envelop glycoprotein GD21. PMID- 12774932 TI - The kinetics of immune reconstitution after cord blood transplantation and selected CD34+ stem cell transplantation in children: comparison with bone marrow transplantation. AB - The present study compares immune reconstitution after allogeneic cord blood transplantation (CBT) and CD34+ stem cell transplantation (CD34-SCT) with that after bone marrow transplantation (BMT). Eighty-eight children who underwent CBT (20 patients), BMT (58), and CD34-SCT (10) were enrolled, and lymphocytes and T-, B-, and natural killer-lymphocyte subsets were monitored for more than 5 years after transplantation. CBT recipients showed significant ircreases in (1) total lymphocyte counts (P < .001), (2) CD4+/CD8+ cell ratios (P < .01), (3) CD4+ and CD4+CD45RA+ cells (P < .001), (4) CD8+CD11b+ cells (P < .001), and (5) CD19+ and CD19+CD5+ cells (P < .0001) and marked decreases in the frequencies of CD8+ and CD8+CD11b- cells (P < .0001). CD34-SCT recipients showed lower lymphocyte counts in the first 6 months and an emergence of lymphocyte and CD4+CD45RA+ cells at approximately 9 months and 1 year. Both CBT and CD34-SCT recipients showed increased frequencies of CD56+ cells at 1 month (CD34-SCT versus BMT, P < .001) but decreased frequencies after 6 months (CBT versus BMT, P < .001). Lymphoproliferative responses to exogenous interleukin 2 were constantly lower in CBT and CD34-SCT recipients than in BMT recipients. These results suggest that the delay in immune reconstitution after CBT in the early phase was mainly qualitative and related to the immaturity of cells, whereas the delay in CD34-SCT was mainly quantitative in the first several months. PMID- 12774935 TI - Antithrombotic effects of magnesium sulfate in in vivo experiments. AB - In this study, magnesium sulfate was effective in reducing the mortality of adenosine diphosphate-induced acute pulmonary thromboembolism in mice, when it was administered intravenously at doses of 100 and 200 microg/g body weight. In addition, intravenous injections of magnesium sulfate (100 and 200 microg/g) significantly prolonged bleeding time in the severed mesenteric arteries of rats by approximately 1.7- and 1.9-fold, respectively, compared with normal saline. Continuous infusion of magnesium sulfate (20 microg/g per minute) for 10 minutes also significantly increased the bleeding time by approximately 1.7-fold, and the bleeding time returned to baseline within 60 minutes of cessation of magnesium sulfate infusion. On the other hand, platelet thrombi formation was induced by irradiating mesenteric venules with filtered light in mice pretreated with intravenous fluorescein sodium. When magnesium sulfate was administered at 300 microg/g during induction of platelet plug formation with 10 microg/kg fluorescein sodium, occlusion time was not significantly prolonged, but a dose of 600 microg/g did significantly prolong the occlusion time. Furthermore, aspirin (250 microg/g) also showed a similar activity in this experiment in prolonging the occlusion time. In conclusion, these results suggest that magnesium sulfate has an effective antithrombotic activity in vivo, and treatment with magnesium sulfate may lower the risk of thromboembolic-related disorders. PMID- 12774936 TI - Immunophenotypic analysis of various blastic crises in chronic myeloid leukemia: correlations between CD7 expression and response to chemotherapy. PMID- 12774937 TI - Supramolecular assemblies of DNA delivery systems. AB - Gene delivery into cultured cells or in vivo is a promising approach to the treatment of diseases. Several gene delivery systems have been developed to promote gene expression either in vitro or in vivo. Concerns about viral-induced immune responses, the risk associated with replication-competent viruses, and production issues have stimulated efforts toward the development of alternative gene delivery systems such as cationic lipids and polymers. These positively charged molecules interact through electrostatic forces with DNA. This results in the formation of highly organized supramolecular structures where DNA molecules are condensed and protected against DNAses degradation. Association of DNA with cationic lipids under a micellar or liposomal form leads to lamellar organization with DNA molecules sandwiched between lipid bilayers. Although the lamellar phase is the common described structure, as evidenced by small-angle X-ray scattering and electron microscopy, monovalent cationic lipid combined with a hexagonal forming lipid resulted with DNA in an inverted hexagonal structure. Despite a lot of effort, the mechanism of gene transfer with cationic carrier is still ill defined. Therefore, correlations need to be established between physicochemical properties of synthetic DNA delivery systems and in vitro and in vivo transfection efficiency. PMID- 12774938 TI - Engineering the surface properties of synthetic gene delivery vectors. AB - Synthetic gene delivery vehicles are a highly promising approach to gene delivery; however, several problems must still be overcome before they can begin to enjoy dinical success. A number of these problems can be addressed by engineering and optimizing the properties of the vector surface, the component of the particle that interacts and "communicates" with tissues and cells during the delivery process. Surfaces must be engineered to satisfy two ostensibly conflicting constraints: the ability to interact specifically with a target cell while avoiding nonspecific protein interactions, particularly with components of the immune system. We summarize progress that has been made in both these areas and discuss several approaches where the intersection of biological and chemical solutions promises to significantly advance the engineering of synthetic vehicles. PMID- 12774939 TI - Histidine containing peptides and polypeptides as nucleic acid vectors. AB - Nucleic acid transfer in mammalian cels is drastically improved with devices which increase their delivery in the cytosol upon endocytosis. In this chapter, we describe the effect on plasmid DNA (pDNA) and oligonucleotide (ODN) transfer, of an histidine-rich peptide (H5WYG), histidylated oligolysine (HoK), and histidylated polylysine (HpK) designed on the basis of the membrane destabilization capacity of poly-L-histidine at a pH dose to that of the endosomes. We report that H5WYG, which permeabilizes the cell membrane at pH 6.4, favors the transfection mediated by lactosylated polylysine/pDNA complexes and, by lowering the pH of extracellular medium, allows the loading of the cytosol and the cell nucleus with ODN. We show that HoK forms small cationic spherical particles of 35 nm with ODN and HpK rod or toroid cationic particles of 100 nm with pDNA. PEGylation stabilizes these particles at physiological salt concentration. We also show that (i) HoK/ODN complexes yield a more than 20-fold increase of the biological activity of antisense ODN towards the inhibition of transient as well as constitutive gene expression and (ii) HpK/pDNA complexes yield a transfection efficiency of 3-4.5 order of magnitude higher than do polylysine/pDNA complexes. We also provide evidence that the effect of these polyhistidylated molecules is mediated by imidazole protonation in endosomes. Overall our data show that polyhistidylated molecules constitute interesting devices for an efficient cytosolic delivery of nucleic acids, and that ionic complexes between histidylated polylysine and a pDNA are attractive for developing a nonviral gene delivery system. PMID- 12774940 TI - Intracellular targets for DNA delivery: nuclei and mitochondria. AB - All discussions on the intracellular delivery of DNA are based on a seemingly evident assumption that the key task is to bring the intact DNA into the cell cytoplasmic compartment, and then the DNA will find its way to a right place. The nuclear genome is usually considered to be this "right place." However, until recently, in numerous experiments on the intracellular DNA delivery, it has been almost completely neglected that cells contain another genome, the mitochondrial one. And, in many cases, this genome should become a therapeutic target. Being delivered inside the cell, DNA actually has two ways to go--to nuclei and to mitchondria, and the proper choice between these two ways may be decisive for the success of gene therapy. Certainly, nuclear DNA delivery is far more advanced than mitochondrial delivery one. In addition, free DNA from the cytoplasm has a strong tendency to spontaneously associate with the nuclear genome. Mitochondria as a target for DNA have much less accessibility, still remaining an important site to reach. Whereas the nuclear delivery of DNA is under active investigation and just awaits better protocols to be elaborated, practically applicable mitochondrial DNA delivery is at its early stage and must be developed almost from scratch. In our studies on intracellular DNA delivery, we have attempted to develop new protocols for targeting DNA to nuclei and to mitochondria. In this chapter we provide a brief description of our recent experiments in both of these important areas. PMID- 12774941 TI - Mannose receptor-mediated gene delivery into antigen presenting dendritic cells. AB - Dendritic cells are professional antigen presenting cells and are unique in their ability to prime naive T cells. Gene modification of dendritic cells is of particular interest for immunotherapy of diseases where the immune system has failed or is aberrantly regulated, such as in cancer or autoimmune disease, respectively. Dendritic cells abundantly express mannose receptor and mannose receptor-related receptors, and receptor-mediated gene transfer via mannose receptor offers a versatile tool for targeted gene delivery into these cells. Accordingly, mannose polyethylenimine DNA transfer complexes were generated and used for gene delivery into dendritic cells. Mannose receptor belongs to the group of scavenger receptors that allow dendritic cells to take up pathogenic material, which is directed for degradation and MHC class II presentation. Therefore, a limiting step of transgene expression by mannose receptor-mediated gene delivery is endosomal degradation of DNA. Several strategies have been explored to overcome this limitation including the addition of endosomolytic components to DNA transfer complexes like adenovirus particles and influenza peptides. Here, we review the current understanding of mannose receptor-mediated gene delivery into dendritic cells and discuss strategies to identify appropriate endosomolytic agents to improve DNA transfer efficacy. PMID- 12774942 TI - Electro-gene-transfer: a new approach for muscle gene delivery. AB - Gene transfer into skeletal muscle cells by direct injection of naked plasmid DNA results in sustained gene expression. Intramuscular injection of plasmid DNA might thus be used to correct myopathies, to secrete locally or systematic therapeutic proteins and to elicit an immune response against specific antigens. However, the potential utility of this technique for gene application in humans is limited by the poor transduction efficiency and the low and highly variable level of gene expression. Different methods are thus being developed to increase the efficiency of gene transfer in muscles. It has been recently reported that a dramatic improvement of DNA transfer is achieved by applying an electric field to the muscle fibers subsequent to local DNA injection. Electro-gene-transfer increases gene expression by several orders of magnitude and strongly reduces interindividual variability. Electroinjection of genes encoding for secreted proteins resulted in sustained expression and disease correction in animal models of gene therapy. Moreover, the immunogenicity of DNA vaccines is dramatically increased when antigen-encoding plasmids are delivered by this technique. This technique may thus have broad and important applications in human gene therapy. This review provides a brief overview of the theory of electro-gene-transfer and describes parameters governing its efficiency in muscle. We also summarize the results obtained with electro-gene-transfer in animal models to date and the technical issues that must be solved before its use for human therapy can be considered. PMID- 12774943 TI - Tumor-targeted gene transfer with DNA polyplexes. AB - Systemic gene delivery systems are needed for therapeutic applications; in some situations, target cells might be spread throughout the organism, as in the case of cancer metastases, which can be reached only via the systemic route. Within the class of nonviral vectors, polymer-based transfection particles named DNA polyplexes and lipid-based systems named DNA lipoplexes are being developed for this purpose. For systemic circulation, masking the surface charge of DNA complexes has to be accomplished to avoid interactions with plasma components, erythrocytes, and the reticuloendothelial system. Among other vector formulations, polyplexes based on polyethylenimine (PEI), shielded with polyethylene glycol (PEG), and linked to the receptor binding ligands transferrin (Tf) or epidermal growth factor (EGF) have been developed. Complexes were found to mediate efficient gene transfer into tumor cell lines in a receptor-dependent and cell-cycle-dependent manner. Systemic administration of surface-shielded Tf PEI polyplexes into the tail vein of mice resulted in preferential gene delivery into distantly growing subcutaneous tumors. In contrast, application of positively charged PEI polyplexes directed gene transfer primarily to the lung. PMID- 12774944 TI - Photochemical transfection: a technology for efficient light-directed gene delivery. AB - Most synthetic gene delivery vectors are taken up in the cell by endocytosis, and inefficient escape of the transgene from endocytic vesicles often is a major barrier for gene transfer by such vectors. To improve endosomal release we have developed a new technology, named photochemical internalization (PCI). PCI is based on photochemical reactions initiated by photosensitizing compounds localized in endocytic vesicles, inducing rupture of these vesicles upon light exposure. PCI constitutes an efficient light-inducible gene transfer method in vitro, which potentially can be developed into a site-specific method for gene delivery in in vivo gene therapy. In this paper the principle behind the PCI technology and the effect of PCI on transfection with different synthetic gene delivery vectors are reviewed. PCI treatment by the photosensitizer aluminum phthalocyanine (AlPcS2a) strongly improves transfection mediated by cationic polymers (e.g., poly-L-lysine and polyethylenimine), while the effect on transfection with cationic lipids is more variable. The timing of the light treatment relative to the transfection period was also important, indicating that release of the DNA from early endosomes is important for the outcome of PCI induced transfection. The possibilities of using PCI as a technology for efficient, site-specific gene delivery in in vivo gene therapy is discussed. PMID- 12774945 TI - Sonoporation: mechanical DNA delivery by ultrasonic cavitation. AB - Development of nonviral gene transfer methods would be a valuable addition to the gene-therapy armamentarium, particularly for localized targeting of specific tissue volumes. Ultrasound can produce a variety of nonthermal bioeffects via acoustic cavitation including DNA delivery. Cavitation bubbles may induce cell death or transient membrane permeabilization (sonoporation) on a single cell level, as well as microvascular hemorrhage and disruption of tissue structure. Application of sonoporation for gene delivery to cells requires control of cavitation activity. Many studies have been performed using in vitro exposure systems, for which cavitation is virtually ubiquitous. In vivo, cavitation initiation and control is more difficult, but can be enhanced by cavitation nucleation agents, such as an ultrasound contrast agent. Sonoporation and ultrasonically enhanced gene delivery has been reported for a wide range of conditions including low frequency sonication (kilohertz frequencies), lithotripter shockwaves, HIFU, and even diagnostic ultrasound (megahertz frequencies). In vitro, a variety of cell lines has been successfully transfected, with concomitant cell killing. In vivo, initial applications have been to cancer gene therapy, for which cell killing can be a useful simultaneous treatment, and to cardiovascular disease. The use of ultrasound for nonviral gene delivery has been demonstrated for a robust array of in vitro and mammalian systems, which provides a fundamental basis and strong promise for development of new gene therapy methods for clinical medicine. PMID- 12774946 TI - Exercise and its troubles. PMID- 12774947 TI - Anticonvulsant prophylaxis for patients with brain tumours: insights from clinical trials. PMID- 12774948 TI - Progress in clinical neurosciences: sepsis-associated encephalopathy: evolving concepts. AB - Systemic sepsis commonly produces brain dysfunction, sepsis-associated encephalopathy, which can vary from a transient, reversible encephalopathy to irreversible brain damage. The encephalopathy in the acute phase clinically resembles many metabolic encephalopathies: a diffuse disturbance in cerebral function with sparing of the brain stem. The severity of the encephalopathy, as reflected in progressive EEG abnormalities, often precedes then parallels dysfunction in other organs. Recent research has revealed a number of potentially important, non-mutually exclusive, mechanisms that have therapeutic implications. PMID- 12774949 TI - Prophylactic anticonvulsants in patients with brain tumour. AB - OBJECTIVE: We conducted a clinical trial to determine if prophylactic anticonvulsants in brain tumour patients (without prior seizures) reduced seizure frequency. We stopped accrual at 100 patients on the basis of the interim analysis. METHODS: One hundred newly diagnosed brain tumour patients received anticonvulsants (AC Group) or not (No AC Group) in this prospective randomized unblinded study. Sixty patients had metastatic, and 40 had primary brain tumours. Forty-six (46%) patients were randomized to the AC Group and 54 (54%) to the No AC Group. Median follow-up was 5.44 months (range 0.13-30.1 months). RESULTS: Seizures occurred in 26 (26%) patients, eleven in the AC Group and 15 in the No AC Group. Seizure-free survivals were not different; at three months 87% of the AC Group and 90% of the No AC Group were seizure-free (log rank test, p = 0.98). Seventy patients died (unrelated to seizures) and survival rates were equivalent in both groups (median survival = 6.8 months versus 5.6 months, respectively; log rank test, p = 0.50). We then terminated accrual at 100 patients because seizure and survival rates were much lower than expected; we would need > or = 900 patients to have a suitably powered study. CONCLUSIONS: These data should be used by individuals contemplating a clinical trial to determine if prophylactic anticonvulsants are effective in subsets of brain tumour patients (e.g. only anaplastic astrocytomas). When taken together with the results of a similar randomized trial, prophylactic anticonvulsants are unlikely to be effective or useful in brain tumour patients who have not had a seizure. PMID- 12774950 TI - The epidemiology of traumatic spinal cord injury in Alberta, Canada. AB - OBJECTIVES: To describe the incidence and pattern of traumatic spinal cord injury and cauda equina injury (SCI) in a geographically defined region of Canada. METHODS: The study period was April 1, 1997 to March 31, 2000. Data were gathered from three provincial sources: administrative data from the Alberta Ministry of Health and Wellness, records from the Alberta Trauma Registry, and death certificates from the Office of the Medical Examiner. RESULTS: From all three data sources, 450 cases of SCI were identified. Of these, 71 (15.8%) died prior to hospitalization. The annual incidence rate was 52.5/million population (95% CI: 47.7, 57.4). For those who survived to hospital admission, the incidence rate was 44.3/million/year (95% CI: 39.8, 48.7). The incidence rates for males were consistently higher than for females for all age groups. Motor vehicle collisions accounted for 56.4% of injuries, followed by falls (19.1%). The highest incidence of motor vehicle-related SCI occurred to those between 15 and 29 years (60/million/year). Fall-related injuries primarily occurred to those older than 60 years (45/million/year). Rural residents were 2.5 times as likely to be injured as urban residents. CONCLUSION: Prevention strategies for SCI should target males of all ages, adolescents and young adults of both sexes, rural residents, motor vehicle collisions, and fall prevention for those older than 60 years. PMID- 12774951 TI - Linkage to the CCM2 locus and genetic heterogeneity in familial cerebral cavernous malformation. AB - BACKGROUND: Cerebral cavernous malformation (CCM) is a form of intracranial vascular disease that may arise sporadically or be dominantly inherited. Linkage studies have revealed genetic heterogeneity among the dominantly inherited forms suggesting the existence of at least three loci called CCM1, CCM2 and CCM3. METHODS: In the present study, we screened five families with dominantly inherited CCM for CCM1 gene mutations with denaturing high performance liquid chromatography (DHPLC). Then, we performed linkage analysis and haplotyping on these five families using highly polymorphic markers at the candidate CCM loci. RESULTS: None of the five families tested with DHPLC were found to have mutations in the CCM1 gene. Based on haplotyping, we identified three families segregating alleles for CCM2, while two families segregated alleles for CCM3. Using linkage analysis, we could confirm that one family (IFCAS-1) had a positive Lod score of 2.03 (p<0.0001) at the CCM2 locus using marker D7S678. CONCLUSIONS: The present study is the first one to replicate linkage at the CCM2 locus and provides a fifth family identified as such. It also supports the concept of genetic heterogeneity in CCM, identifying four other families that showed no mutations in the CCM1 gene. PMID- 12774952 TI - Quantitative motor assessment in myotonic dystrophy. AB - OBJECTIVE: To establish baseline data, using a quantitative motor evaluation protocol, prior to a prospective longitudinal study of the natural history of muscular involvement in myotonic dystrophy (DM). DESIGN/METHODS: We conducted a cross-sectional study using a protocol consisting of manual muscle testing (MMT), quantitative muscle testing (QMT), and timed functional testing (TFT) on 50 definite DM patients (27 men, 23 women), aged 16 to 67 years. The relationships between MMT, QMT and TFT scores and disease duration were examined using linear regression analysis. RESULTS: The muscle weakness was symmetric and the neck flexors and the distal muscles of upper and lower extremities were weaker than proximal muscles. Using MMT scores, the average strength decline was 0.95% per year and was similar for men and women. The strength decline was significantly more rapid for distal muscles than for proximal muscles. Quantitative muscle testing scores documented a strength decline per year of disease duration of 1.2 1.6% for the hip flexors and of 2.0-3.0% for the hand grip flexors. CONCLUSIONS: We observed significant linear relationships between the scores generated by this protocol and disease duration. These data illustrate the distal to proximal progression of muscular involvement in DM, a pattern of progression well recognized by the clinicians. The follow-up assessment of a large DM cohort in a longitudinal study will establish whether this quantitative protocol provides sensitive measures of the disease progression. PMID- 12774953 TI - Short term predictors of unemployment in multiple sclerosis patients. AB - BACKGROUND: Unemployment is common in people with multiple sclerosis (MS) and is associated with loss of income and impaired health related quality of life. This study determined variables associated with unemployment and risk factors for the development of unemployment in people with MS. METHODS: Ninety-six patients who were under age 65 and participated in two previous studies to measure economic costs and health related quality of life in MS were included. The baseline employment rate and variables associated with unemployment at baseline were determined. The ability of these variables to predict unemployment over the next two and a half years was then evaluated. RESULTS: At baseline 50.1% (50/96) of participants were employed. Two and a half years later only 40.6% (39/96) remained employed. This represents loss of employment for 22.0% (11/50) of those originally employed. Factors associated with unemployment at baseline included greater disability, progressive disease course, longer disease duration, and older age. Risk factors for loss of employment over the next 2.5 years included greater disability and older age. CONCLUSIONS: This study confirms the low employment rate among people with MS and confirms the association of several previously-reported factors with greater risk of unemployment. It is also the first study to confirm that some of these factors also increase the risk of future unemployment. People with MS who are over age 39 or have moderate disability and are still employed can now be identified as at risk for becoming unemployed over the next 2.5 years. They should be considered for interventions to maintain employment or to lessen the impact of unemployment. PMID- 12774954 TI - Lazaroid attenuates edema by stabilizing ATPase in the traumatized rat brain. AB - OBJECTIVE: The aim of the present study was to determine the potential therapeutic value of the lazaroid U-83836E on blood brain barrier (BBB) breakdown and edema with respect to the changes in the synaptosomal Na+/K+ and Mg(2+)/Ca(2+)-adenosinetriphosphatase (ATPase) activities, tissue malondialdehyde levels and the neuronal viability in the rat brain subjected to cerebral trauma. METHODS: Traumatic brain injury (TBI) was introduced by applying a 75 gm. cm force to the right parietal cortex using the weight-drop method. The first set of animals was used for determining time course changes of the synaptosomal Na+/K+ and Mg(2+)/Ca(2+)-ATPase and the malondialdehyde levels and were sacrificed 2, 6 and 24h after lesion production. A group of the animals was treated with U-83836E proir to TBI and sacrificed 24h after cerebral injury. A second set of animals was used for evaluating the alterations in BBB disruption and tissue water content and were sacrificed 2, 6 and 24h after lesion production. Two groups of animals were treated with U-83836E and sacrificed after 2 and 24h following TBI. U-83836E was given intraperitoneally thirty minutes before trauma at a dose of 10 mg/kg. Neuronal necrosis was also evaluated in the groups of U-83836E and physiological saline-treated animals. RESULTS: Extravasation of Evans blue into the traumatized hemisphere was maximum at 2h (p<0.001) and returned close to the control levels at 24h after TBI (p>0.05). Edema had developed progressively over time and reached the maximum degree of 2.1% (p<0.001) at 24h. U-83836E showed no effect on the BBB breakdown and the tissue water content at 2h and still had no effect on the BBB breakdown after 24h following the trauma (p>0.05), although it reduced edema after 24h (p<0.01). The losses of Na+/K+ and Mg(2+)/Ca(2+)-ATPase activities were found as 39.5% (p<0.001) and 29.4% (p<0.01) of the control value, respectively, and remained at the decreased levels throughout the experiment. Malondialdehyde level continued to increase over time reaching up to 209% (p<0.001) of the control value 24h after TBI. Both ATPase activities were improved to near control values (p>.05) by the effect of U-83836E. U-83836E inhibited the increase of lipid peroxidation (p<0.001) and also salvaged neuronal necrosis (p<0.05). CONCLUSION: U-83836E given prophylactically after cerebral trauma appears to reduce edema, possibly by inhibiting increases in lipid peroxidation and by stabilizing ATPase. Further studies are recommended to verify the similar effects of the brain penetrating lazaroids when they are given after trauma. PMID- 12774955 TI - Spontaneous extra-cranial vertebral artery dissection. PMID- 12774956 TI - Magnetic resonance image findings and surgical considerations in T1-2 disc herniation. AB - OBJECTIVE: To report a case of disc herniation at T1-2. CLINICAL PRESENTATION: A 57-year-old man presented with hand weakness, Horner's syndrome, and pain radiating along the medial aspect of one upper extremity. Magnetic resonance imaging demonstrated extruded T1-2 disc herniation with upward herniation of a sequestrated fragment. INTERVENTION: An anterior approach was used to excise the disc, that was compressing the spinal cord and the T1 nerve root. All the patient's symptoms resolved completely, including Horner's syndrome. CONCLUSION: Anterior discectomy may be the simplest and most effective method for disc excision and relief of spinal cord and T1 nerve root compression. PMID- 12774957 TI - Acute disseminated encephalomyelitis associated with Pasteurella multocida meningitis. AB - OBJECTIVE: To describe a case of Pasteurella multocida meningitis associated with acute disseminated encephalomyelitis (ADEM). CASE REPORT: A 33-year-old woman employed in a dog pound presented herself to hospital with fever and meningismus and was found to have culture positive Pasteurella multocida meningitis. Despite appropriate antibiotic treatment her clinical course was characterized by a persistent fever and worsening encephalopathy, which prompted further neurological investigation. Spinal fluid exam and serial MRI scans as well as her one-year clinical course were found to be compatible with ADEM. CONCLUSION: Persistent fever and worsening encephalopathy in meningitis may indicate a para infectious immune process such as ADEM, and may serve as indications for further neurological investigation. PMID- 12774958 TI - Basilar artery stenosis mimicking the lacunar syndrome of pure motor hemiparesis. AB - BACKGROUND: Stereotyped, repeated transient ischemic attacks manifesting as pure motor hemiparesis are most often attributed to ischemia of the internal capsule or ventral pons resulting from in situ disease of the small penetrating arteries. CASE DESCRIPTION: We report a 61-year-old man presenting with recurrent episodes of left-sided weakness consistent with the lacunar syndrome of pure motor hemiparesis. Subsequent neuroimaging revealed infarction of the right ventral pons and a critical basilar artery stenosis as the inciting lesion. Despite maximal antithrombotic therapy, he continued to have repeated symptoms. Angioplasty and stenting were attempted but both failed due to plaque recoil and technical difficulties. After the procedure, the patient had no further ischemic episodes and remained symptom-free at two months. CONCLUSIONS: This case illustrates the imprecise and discordant relationship between the mode of presentation of a stroke syndrome and its presumed pathophysiology. The lacunar syndrome of pure motor hemiparesis should be recognized by clinicians as a mode of stroke presentation due not only to small vessel disease, but also to large artery atherosclerotic disease such as basilar artery stenosis. Prompt institution of treatment can lead to a good clinical outcome. PMID- 12774959 TI - Dopamine transporter binding in Wilson's disease. AB - INTRODUCTION: In Wilson's disease (WD), brain magnetic resonance images (MRI) show increased signal intensity in T2 weighted images in the lenticular nuclei, thalamus and brainstem, including the substantia nigra. A poor therapeutic response to levodopa in WD suggests the mechanism of a postsynaptic abnormality. However positron emission tomography studies show an involvement of the nigrostriatal presynaptic dopaminergic pathway. CASE REPORT: We report the clinical manifestations in a case of WD with akinetic-rigid syndrome and initial hesitation. The brain MRI showed an increased signal intensity lesion in the substantia nigra region, in addition to basal ganglion and thalamic lesions. However, dopamine transporter (DAT) imaging with 99mTc-TRODAT-1 revealed a nonsignificantly increased DAT uptake, suggesting a normal presynaptic nigrostriatal dopaminergic terminal. CONCLUSION: We suggest that significant heterogeneity can be found in WD patients and a normal presynaptic dopaminergic pathway may occur in some patients, even those with typical akinetic-rigid syndrome and evidence of substantia nigra involvement in the brain on MRI. PMID- 12774960 TI - Stroke secondary to post-partum coronary artery dissection. AB - BACKGROUND: There is an increased risk of stroke during pregnancy and the puerperium. This risk is greatest in the post-partum period. Embolic sources secondary to cardiac abnormalities and cerebral arterial dissection are recognized as potential etiologies. CASE: A 32-year-old woman presented with left sided weakness eight days post-partum. Imaging studies identified infarction in the territory of the right middle cerebral artery. Cardiac investigations revealed acute myocardial infarction secondary to dissection of the left main coronary artery. The patient underwent coronary artery bypass grafting with significant recovery of cardiac function. At two years post-presentation, her neurological deficits have resolved. DISCUSSION: Myocardial infarction secondary to coronary artery dissection has been long-recognized. Cerebral arterial dissection in association with pregnancy is also well-documented. This is a rare case of a patient presenting with stroke secondary to acute myocardial infarction secondary to coronary artery dissection. PMID- 12774961 TI - What is causing crescendo transient ischemic attacks? AB - BACKGROUND AND METHODS: We discuss a case report from a patient who had symptoms of transient neurological deficits in the presence of a chronic subdural hematoma and severe carotid stenosis. Multi-modality imaging was used to guide management. RESULTS: The symptoms settled without carotid intervention and were presumed due to the subdural hematoma. CONCLUSIONS: Severe symptomatic carotid stenosis is treated with carotid endarterectomy. In some patients with transient neurological deficits, the diagnosis is not as simple as first thought. Multi-modality imaging (MRI, TCD and CT) can help differentiate the causative lesion. PMID- 12774962 TI - Re: Perioperative ulnar neuropathies; a medical legal review. John D. Stewart, Stephen H. Shantz. Can J Neurol Sci 2003; 30: 15-19. PMID- 12774963 TI - Re: Tests of motor function in patients suspected of having mild unilateral cerebral lesions. Teitelbaum JS, Eliasziw M, Garner M. Can J Neurol Sci 2002; 29: 337-344. PMID- 12774964 TI - NSAID-induced gastrointestinal adverse effects in dogs--can we avoid them? PMID- 12774965 TI - Cardiac output technologies with special reference to the horse. AB - Critical illness, anesthesia, primary cardiovascular disease, and exercise may result in marked hemodynamic alterations. Measuring cardiac output (CO) is central to defining these alterations for both clinician and researcher. In the past 10 years, several new methods of measuring CO have been developed for the human medical market. Some of these methods are now validated in the horse and are in clinical use. The Fick method has been used in equine research for more than a century. It depends on simultaneous measurement of mixed venous (pulmonary arterial) and peripheral arterial oxygen content and oxygen uptake by the lungs. The technique is technically demanding, which restricts its clinical use. Indicator dilution techniques, with indocyanine green, cold (thermodilution), or lithium as the marker, have also been widely used in the horse. The indocyanine technique is cumbersome, and thermodilution requires right heart catheterization, which is not a benign procedure, making both of these methods less than ideal for clinical use. Lithium dilution requires catheterization of a peripheral artery and a jugular vein. It has recently been validated in anesthetized adult horses and neonatal foals. Doppler echocardiography is a noninvasive ultrasound-based technique. More accurate measurements are obtained with transesophageal than with transthoracic measurements; however, both methods require considerable technical expertise. Bioimpedance and pulse contour analysis are 2 new methods that have yet to be validated in the horse. With the currently available technology, lithium dilution appears to be the method of measuring CO best suited to the equine clinic. PMID- 12774966 TI - Adjuvants in veterinary vaccines: modes of action and adverse effects. AB - Vaccine adjuvants are chemicals, microbial components, or mammalian proteins that enhance the immune response to vaccine antigens. Interest in reducing vaccine related adverse effects and inducing specific types of immunity has led to the development of numerous new adjuvants. Adjuvants in development or in experimental and commercial vaccines include aluminum salts (alum), oil emulsions, saponins, immune-stimulating complexes (ISCOMs), liposomes, microparticles, nonionic block copolymers, derivatized polysaccharides, cytokines, and a wide variety of bacterial derivatives. The mechanisms of action of these diverse compounds vary, as does their induction of cell-mediated and antibody responses. Factors influencing the selection of an adjuvant include animal species, specific pathogen, vaccine antigen, route of immunization, and type of immunity needed. PMID- 12774967 TI - The effect of dosing interval on the efficacy of misoprostol in the prevention of aspirin-induced gastric injury. AB - The effect of twice-daily administration of misoprostol on aspirin-induced gastric injury was evaluated. Twenty-four random-source dogs were divided into groups that received aspirin and misoprostol as follows: group I, aspirin 25 mg/kg PO q8h and placebo PO q8h; group II, aspirin 25 mg/kg PO q8h and misoprostol 3 microg/kg PO q8h; group III, aspirin 25 mg/kg PO q8h, misoprostol 3 microg/kg PO q12h, and placebo PO q24h; and group IV, aspirin 25 mg/kg PO q8h, misoprostol 3 microg/kg PO q24h, and placebo PO q12h for 28 days. Gastroscopy was performed on days -9, 5, 14, and 28. Visible lesions were scored on a scale of 1 (mucosal hemorrhage) to 11 (perforating ulcer). No difference in total score was identified between groups I and IV on any day. Median total scores for groups II and III were significantly (P < or = .05) lower compared to groups I and IV on day 5. Group III had a significantly lower score (P < or = .05) than groups I, II, and IV on day 28. This study suggests that misoprostol 3 microg/kg PO q12h is as effective as misoprostol 3 microg/kg PO q8h in preventing aspirin-induced gastric injury in this model. However, misoprostol 3 microg/ kg PO q8h was less effective in preventing aspirin-induced gastric injury on days 14 and 28 than in previous studies. No difference among numbers of dog-days of vomiting, diarrhea, or anorexia was detected among groups. PMID- 12774968 TI - A scoring index for disease activity in canine inflammatory bowel disease. AB - The clinical course of inflammatory bowel disease (IBD) in dogs is characterized by spontaneous exacerbations and remissions, which makes assessment of disease burden difficult. The objectives of this study were to develop a scoring system for evaluation of canine IBD activity and to validate this scoring method by correlating it to objective laboratory and histologic indices of intestinal inflammation. Fifty-eight dogs with IBD were evaluated prospectively and compared to 9 disease-free control dogs. Clinical disease activity was quantified by a simple scoring system, the canine IBD activity index (CIBDAI), and compared to serum concentrations of C-reactive protein (CRP), haptoglobin (HAP), alpha-acid glycoprotein (AGP), and serum amyloid A (SAA), as well as histology scores derived from endoscopic biopsy specimens. Forty-six dogs were available for a reevaluation of the CIBDAI, CRP HAP, and AGP, and 34 dogs had repeat analysis of SAA performed after medical therapy. Serum concentrations of CRP were significantly (P < .02) increased in dogs with CIBDAI scores > or = 5 (mild disease activity or greater) compared to controls. Among IBD dogs, the CIBDAI showed good correlation (r = 0.82, P < .0001) to both histology and HAP scores, but CRP also was a strong co-correlate of disease activity. The IBD dogs showed significantly (P < .0001) decreased CIBDAI and CRP values but significantly (P < .0001) increased HAP concentrations after medical therapy compared to pretreatment values. We conclude that the CIBDAI is a reliable measure of inflammatory activity in canine IBD and that CRP is suitable for laboratory evaluation of the effect of therapy in these patients. PMID- 12774970 TI - Diazepam as a treatment for metronidazole toxicosis in dogs: a retrospective study of 21 cases. AB - The currently recommended treatment for metronidazole toxicosis is drug discontinuation and supportive therapy. Reported recovery times are 1-2 weeks. The records of 21 dogs with metronidazole toxicosis were retrospectively analyzed to determine whether diazepam improved recovery. The dosage and duration of metronidazole therapy and the response and recovery times of 13 dogs treated with diazepam were compared to those of 8 dogs receiving only supportive care. Response time was defined as the time to resolution of the debilitating clinical signs. Recovery time was the time to resolution of all residual clinical signs. The average dosage and duration of metronidazole administration for the diazepam treated and untreated groups were 60.3 mg/kg/d for 44.9 days and 65.1 mg/kg/d for 37.25 days. The protocol for diazepam administration consisted of an initial i.v. bolus and then diazepam PO q8h for 3 days. The average dosage of both the i.v. and PO diazepam was 0.43 mg/kg. The average response time for the diazepam treated dogs was 13.4 hours compared to 4.25 days for the untreated group. Recovery time also was markedly shorter for the diazepam-treated dogs (38.8 hours) compared to the untreated group (11 days). Results of this study showed that dogs with metronidazole toxicosis recover faster when treated with diazepam. Although the mechanism of metronidazole toxicosis or how diazepam exerts its favorable effect is not known, it is likely related to modulation of the gamma aminobutyric acid (GABA) receptor within the cerebellar and vestibular systems. PMID- 12774969 TI - Percutaneous ultrasound-guided cholecystocentesis in healthy cats. AB - Percutaneous ultrasound-guided cholecystocentesis (PUC) is a minimally invasive technique for bile collection that is used successfully in human patients with cholecystitis. Its use in veterinary medicine for evaluation of hepatobiliary disorders has been limited because of the perceived unacceptable risk of bile peritonitis. An experimental study was conducted to evaluate the safety and efficacy of PUC, to collect data on bile cytology and bacteriologic culture, and to attempt to isolate Helicobacter spp. from the bile of healthy cats. In fasted and sedated cats, PUC was performed with a 22-gauge 1.5-in. (3.81 cm) needle with an attached 12-mL syringe via a right-sided transhepatic approach (n = 1) or into the fundus of the gallbladder (n = 11) via a right ventral abdominal approach. An attempt was made to completely empty the gallbladder. A small amount of abdominal effusion, consistent with bile or blood, was seen ultrasonographically immediately after aspiration in the 1st cat. Ultrasonographic complications with the 2nd technique were not observed in the remaining 11 cats. Decreased appetite and evidence of mild abdominal pain were detected in 4 cats within 2 days after PUC. The mean neutrophil count increased 2 days after PUC (P < .01) but remained within the reference range. The bile was acellular in 11 of 12 cats, and aerobic, anaerobic, and Helicobacter spp. cultures yielded no growth in 12 of 12 cats. There were no remarkable gross or histologic lesions of abdominal organs at the postmortem examination (8 cats) performed 7-8 days after the procedure. PUC appears to be a safe and technically simple procedure. Further studies are warranted to determine the use and safety of PUC in cats with hepatobiliary diseases. PMID- 12774972 TI - Serum nitrate and nitrite in dogs with spontaneous cardiac disease. AB - The purpose of this study was to determine whether nitric oxide (NO) concentrations are high in dogs with chronic valvular disease (CVD) and dilated cardiomyopathy (DCM) compared to healthy controls and to determine whether NO concentrations are correlated with type of cardiac disease, disease severity, medical therapy, or serum tumor necrosis factor (TNF) and interleukin-1 (IL-1). Blood was collected from 32 dogs with DCM, from 10 dogs with CVD, and from 10 healthy controls. Indirect determination of NO concentrations was performed by a commercial photoabsorbance assay that uses a Greiss reagent to measure the concentration of nitrite and nitrate (NN), end products of NO metabolism. TNF and IL-1 activities were measured by bioassay. Mean NN concentrations were significantly higher in dogs with heart disease (median, 4.57 microM; range, 0.00 31.05 microM) than in controls (median, 0.00 microM; range, 0.00-6.16 microM; P = .04). NN concentrations in dogs with cardiac disease were not correlated with type or severity of cardiac disease, medication type, or TNF and IL-1 concentrations. NN concentrations were inversely correlated with fractional shortening. The results of this study suggest that metabolites of NO are increased in some dogs with cardiac disease, but these increases appear to be independent of disease severity, TNF and IL-1 concentrations, and type of pharmacologic intervention. PMID- 12774971 TI - Prevalence of gastric lesions in racing Alaskan sled dogs. AB - Human and equine athletes are reported to have a high prevalence of gastric disease, and anecdotal evidence suggests a similar phenomenon applies to racing sled dogs. To investigate the prevalence of gastric disease in racing sled dogs, we conducted 2 gastroscopy studies on dogs competing in the annual Iditarod Sled Dog Race. A pilot study of dogs that were either dropped from the 2000 Iditarod Sled Dog Race because of illness or that finished the race indicated that, approximately 5 days after competing, 10 of 28 dogs (35%) had endoscopic evidence of gastric ulceration, erosion, or hemorrhage. The next year, an endoscopic study of 73 dogs participating in the 2001 Iditarod race was performed in order to evaluate a larger population of dogs. Data from 70 of these dogs could be used; 34 (48.5%) had ulceration, erosion, gastric hemorrhage, or some combination of these findings. When this group of 70 dogs was compared retrospectively to a control group of 87 dogs presented to the Texas A&M University (TAMU) Veterinary Medical Teaching Hospital, the Iditarod sled dogs had a significantly higher prevalence (P = .049) of gastric lesions. These findings suggest that, similar to athletes of other species, elite canine athletes have an increased prevalence of gastric disease compared to the canine population at large. PMID- 12774973 TI - Clinical characteristics and inheritance of idiopathic epilepsy in Vizslas. AB - Medical record, seizure survey, and telephone interview information was obtained for 29 Vizslas with idiopathic epilepsy (IE), 74 unaffected siblings, and 41 parents to determine the common clinical characteristics and most likely mode of inheritance. IE was diagnosed on the basis of the age of seizure onset, laboratory results, and neurologic examination findings. Computerized tomography (CT) or magnetic resonance imaging (MRI) scan with cerebrospinal fluid (CSF) analysis was required for the inclusion of dogs with an age of seizure onset of < 6 months or > 5 years. Simple segregation analysis was performed with an ascertainment correction and chi-square analysis. IE appeared to be familial in these pedigrees, with 79% of affected Vizslas exhibiting partial onset seizures. Partial seizure signs included a combination of limb tremors, staring, pupillary dilatation, or salivation without loss of consciousness in > 50% of the dogs with partial signs. The estimated segregation frequency of P = .22 (95% CI, P = .08 to .36) was consistent with autosomal recessive inheritance; however, polygenic inheritance could not be excluded as a possibility. Simulated linkage with FASTSLINK estimated that the average logarithm of odds (LOD) score would be 3.23 with a 10-centimorgan (cM) whole-genome scan for these families, indicating that these families would be useful for a whole-genome scan to potentially find the chromosomal segment(s) containing the epilepsy gene or genes. We conclude that IE in Vizslas appears to be primarily a partial onset seizure disorder that may be inherited as an autosomal recessive trait. PMID- 12774974 TI - Feline epitheliotropic intestinal malignant lymphoma: 10 cases (1997-2000). AB - The clinical, histopathologic, and immunohistochemical features of 10 cats with epitheliotropic intestinal malignant lymphoma (EIL) are described. Intestinal biopsy samples were reviewed by 3 pathologists to confirm the diagnosis of EIL. These samples (n = 10) were compared to the intestinal biopsies of normal cats (n = 11), cats with inflammatory bowel disease (IBD; n = 7), and cats with non-EIL (n = 9) for quantification and immunophenotyping of intraepithelial lymphocytes. Immunophenotypic studies were performed with CD3 and CD79a antibody stains to assess for T- and B-cell immunoreactivity, respectively. EIL biopsies had markedly more intraepithelial lymphocytes than normal intestine (NRL) and samples from cats with IBD. However, no marked difference was observed in the number of intraepithelial lymphocytes in cats with non-EIL compared to cats with EIL. Regardless of the histologic diagnosis, the intraepithelial lymphocytes in all cats were small- to intermediate-sized T cells. Clinical findings and imaging studies in the cats identified minimal or nonspecific findings in affected cats. Most cats fit the typical profile of cats with IBD or alimentary malignant lymphoma. Nine of 10 cats with EIL were treated with prednisone with or without additional chemotherapy. Four cats were refractory to chemotherapy and were euthanized within 3.5 months. The remaining 5 cats had long-term survival times of 11 months or greater. The median survival time was 11 months. Additional studies are warranted to better characterize EIL and its relationship to IBD in cats and non-EIL and to identify optimal treatment strategies for this disease. PMID- 12774975 TI - An assessment of the terminology used by diplomates and students to describe the character of equine mitral and aortic valve regurgitant murmurs: correlations with the physical properties of the sounds. AB - Twenty students and 16 diplomates listened to 7 recordings made from 7 horses with either aortic (n = 3) or mitral valve (n = 4) regurgitant murmurs. A total of 30 different terms were used to describe the character of these murmurs. However, only 4 terms were used in a repeatable and consistent manner. Most people described the character of a given mitral or aortic valve murmur with 1 or 2 terms. Diplomates drew from a pool of terms that was about half the size of that used by students--8.1 +/- 2.0 terms for diplomats (mean +/- 1 SD) versus 13.1 +/- 1.8 terms for students (P > .001). Only blowing, honking, buzzing, and musical were markedly associated with the recording played. Frequency analysis of the murmurs allowed them to be classified as containing harmonics (n = 4) or not containing harmonics (n = 3). Blowing was used to describe murmurs without harmonics on 39 of 48 occasions and corresponds to the term noisy used in some older descriptions of equine murmurs. Honking, musical, and buzzing were markedly associated with murmurs that contained harmonics; these terms were used 23, 13, and 12 of a possible 64 times, respectively. The frequency of buzzing and honking murmurs (72.7 +/- 9.3 and 88.4 +/- 46.3 Hz, respectively) was markedly lower than that of musical murmurs (156.8 +/- 81.1 Hz) (all P values < .01). Honking murmurs (0.392 +/- 0.092 seconds) were shorter than those described as buzzing or musical (0.496 +/- 0.205 and 0.504 +/- 0.116 seconds, respectively). The data suggest that the terminology for the character of aortic and mitral regurgitant murmurs should be restricted to 4 terms: blowing, honking, buzzing, and musical. Honking, buzzing, and musical describe murmurs with a peak dominant frequency and harmonics; blowing describes murmurs without a peak frequency. Effective communication could be enhanced by playing examples of reference sounds when these terms are taught so that nomenclature is used more uniformly. PMID- 12774976 TI - Serum IgM concentrations in normal, fit horses and horses with lymphoma or other medical conditions. AB - The purposes of this study were to (1) prospectively establish serum IgM and IgG concentrations in normal, fit, adult horses over time and (2) determine the accuracy of serum IgM concentrations for diagnosing lymphoma. Serial IgM and IgG concentrations were measured with a radial immunodiffusion assay in 25 regularly exercised horses at 6-week intervals. Horses had serum IgM concentrations ranging from 50 to 242 mg/dL over 5 months, with 20% of horses having IgM < or = 60 mg/dL. The normal range for IgM in fit horses should be considered 103 +/- 40 mg/dL and a cut-point for an IgM deficiency, < or = 23 mg/dL. IgG concentrations ranged from 1,372 to 3,032 mg/dL. Retrospectively, medical records of adult horses (n = 103) admitted to the Cornell University Hospital for Animals for which serum IgM was measured were examined. Horses were categorized as "lymphoma negative" (n = 34) or "lymphoma positive" (n = 18). The sensitivity and specificity of a serum IgM concentration (< or = 60 mg/dL) for detecting equine lymphoma was 50 and 35%, respectively. At the new cut-point (< or = 23 mg/dL), the sensitivity was low at 28% and the specificity improved to 88%. The negative predictive values at various population prevalences indicate that a horse with a high serum IgM (> 23 mg/dL) is unlikely to have lymphoma, whereas the positive predictive value (70%) does not allow for reliable determination of lymphoma in a horse with serum IgM < or = 23 mg/dL. Therefore, serum IgM concentrations should not be used as a screening test for equine lymphoma. PMID- 12774977 TI - Prognosis in equine medical and surgical colic. AB - A retrospective study was performed on 649 horses admitted to the Clinic of Equine Internal Medicine (Utrecht University) for gastrointestinal colic. The aim of this study was to provide a better guideline for determining prognosis in horses with colic. Short- and long-term survival were evaluated, and Cox regression analysis was performed to identify the clinical and clinicopathologic variables usable to predict survival. Intestinal displacements and strangulations were the most frequently diagnosed causes of colic. Regarding the surgically treated horses, 54% were discharged and 88% of these were still alive after 1 year. In comparison, 85% of medically treated patients were discharged and 93% were still alive after 1 year. Only 4.4% of all long-term survivors did not return to the expected level of performance, and 32% suffered from recurrent colic. Duration of colic signs, heart rate, intestinal motility, skin tenting, level of pain, and gross appearance of peritoneal fluid were shown to be significantly associated with survival. White blood cell count, packed cell volume, blood pH, and color of the mucous membranes did not show any prognostic significance. Thus, clinical variables appeared to be the most relevant predictors of the outcome of gastrointestinal colic. PMID- 12774978 TI - Transvenous embolization of the patent ductus arteriosus with detachable coils in 2 cats. PMID- 12774979 TI - Ivermectin toxicity in an Australian Shepherd dog with the MDR1 mutation associated with ivermectin sensitivity in Collies. PMID- 12774980 TI - Simultaneously occurring oligodendroglioma and meningioma in a dog. PMID- 12774981 TI - Intravascular hemolysis associated with liver disease in a horse with marked neutrophil hypersegmentation. PMID- 12774982 TI - High cardiac troponin I serum concentration in a horse with multiform ventricular tachycardia and myocardial necrosis. PMID- 12774983 TI - Using continous PM2.5 monitoring data to report an air quality index. PMID- 12774984 TI - Volatile organic profiles and photochemical potentials from motorcycle engine exhaust. AB - This study surveyed emissions from 2- and 4-stroke new and in-use motorcycles. Emission tests were carried out on a dynamometer following the designated test procedure of the Economic Commission for Europe (ECE). Samples were derived during various driving stages, which included idle, acceleration, 30 km/hr cruise, 50 km/hr cruise, and deceleration. All test motorcycles (10 new and 15 in use) complied with Taiwan Environmental Protection Administration's Phase III Motorcycle Emission Standards. The dominant volatile organic carbon (VOC) species were isopentane (53 and 295 mg/km, 2- and 4-stroke, respectively), 2 methylpentane (75 and 83 mg/km), 3-methylpentane (34 and 66 mg/km), and toluene (30 and 100 mg/km). The VOC emission factors for the 2-/4-stroke motorcycles were 311/344 (new) and 1479/433 (in-use) mg/km, respectively. In addition, the dominant carbonyl species for the new and in-use motorcycles were formaldehyde (0.4 and 0.7 mg/km, respectively), acetaldehyde (0.6 and 1.2 mg/km), and acetone (0.5 and 0.7 mg/km). The carbonyl compound emission factors for the 2- and 4 stroke motorcycles were 3.2/3.1 (new) and 5.3/4.6 (in-use) mg/km, respectively. The ozone formation potentials, based on an ECE test cycle, show that the values from the in-use motorcycles were higher than those from the new motorcycles. The dominant VOC species for the ozone formation potential were propylene (65 and 502 mg-O3/km, respectively), isopentane (98 and 501 mg-O3/km), 2-methylpentane (152 and 167 mg-O3/ km), 3-methylpentane (79 and 253 mg-O3/km), and toluene (127 and 398 mg-O3/km). Further, the dominant carbonyl species were formaldehyde (4.1 and 6.2 mg-O3/ km, new and in-use, respectively) and acetaldehyde (4.8 and 9 mg O3/km). PMID- 12774985 TI - Variables affecting emissions of PCDD/Fs from uncontrolled combustion of household waste in barrels. AB - The uncontrolled burning of household waste in barrels has recently been implicated as a major source of airborne emissions of polychlorinated dibenzo-p dioxins and polychlorinated dibenzofurans (PCDD/Fs). A detailed, systematic study to understand the variables affecting emissions of PCDD/Fs from burn barrels was performed. The waste composition, fullness of the barrel, and the combustion conditions within the barrel all contribute significantly to determining the emissions of PCDD/Fs from burn barrels. The study found no statistically significant effect on emissions from the Cl content of waste except at high levels, which are not representative of typical household waste. At these elevated Cl concentrations, the impact of Cl on PCDD/F emissions was found to be independent of the form of the Cl (inorganic or organic). For typical burn conditions, most of the PCDD/F emissions appear to be associated with the later stages of the burn when the waste is smoldering. Polychlorinated biphenyls (PCBs) were also measured for a subset of the tests. For the nominal waste composition, the average emissions were 76.8 ng toxic equivalency units (TEQ)WHO98/kg of waste combusted, which suggests that uncontrolled burning of household waste could be a major source of airborne PCDD/Fs in the United States. PMID- 12774986 TI - Prediction of SO2 levels using neural networks. AB - In this paper, we present an adaptation of the air pollution control help system in the neighborhood of a power plant in As Pontes (A Coruna, Spain), property of Endesa Generacion S.A., to the European Council Directive 1999/30/CE. This system contains a statistic prediction made half an hour before the measurement, and it helps the staff in the power plant prevent air quality level episodes. The prediction is made using neural network models. This prediction is compared with one made by a semiparametric model. PMID- 12774987 TI - A two-stage interval-stochastic programming model for waste management under uncertainty. AB - This study introduces a two-stage interval-stochastic programming (TISP) model for the planning of solid-waste management systems under uncertainty. The model is derived by incorporating the concept of two-stage stochastic programming within an interval-parameter optimization framework. The approach has the advantage that policy determined by the authorities, and uncertain information expressed as intervals and probability distributions, can be effectively communicated into the optimization processes and resulting solutions. In the modeling formulation, penalties are imposed when policies expressed as allowable waste-loading levels are violated. In its solution algorithm, the TISP model is converted into two deterministic submodels, which correspond to the lower and upper bounds for the desired objective-function value. Interval solutions, which are stable in the given decision space with associated levels of system-failure risk, can then be obtained by solving the two submodels sequentially. Two special characteristics of the proposed approach make it unique compared with other optimization techniques that deal with uncertainties. First, the TISP model provides a linkage to predefined policies determined by authorities that have to be respected when a modeling effort is undertaken; second, it furnishes the reflection of uncertainties presented as both probabilities and intervals. The developed model is applied to a hypothetical case study of regional solid-waste management. The results indicate that reasonable solutions have been generated. They provide desired waste-flow patterns with minimized system costs and maximized system feasibility. The solutions present as stable interval solutions with different risk levels in violating the waste-loading criterion and can be used for generating decision alternatives. PMID- 12774988 TI - Ground-level ozone mapping in large urban areas using multivariate statistical analysis: application to the Sao Paulo Metropolitan Area. AB - A statistical study on the behavior of ground-level O3 concentration in different regions of a large urban area was carried out, with emphasis on pollutant gas concentrations and meteorological variables. The study was based on data generated by a network of measuring stations distributed throughout the Sao Paulo Metropolitan Area, in regions with different characteristics of traffic and economic activities. The combined application of principal component analysis and clustering techniques to data collected from 1997 until 2000 has led to the identification of implicit relationships between variables that have been associated with dominant processes related to O3 formation in different locations. Similarities between different regions of the city have also been detected and associated with local characteristics. The results indicate that the application of such statistical techniques to data collected in large urban areas enables the grouping of different regions according to their behavior in terms of O3 levels, as well as the identification of dominant processes in each group. These techniques are thus important in the planning of air pollution policies, especially in the case of O3, a pollutant that is not directly related to pollution levels alone. PMID- 12774989 TI - Residential indoor, outdoor, and workplace concentrations of carbonyl compounds: relationships with personal exposure concentrations and correlation with sources. AB - Personal 48-hr exposures of 15 randomly selected participants as well as microenvironment concentrations in each participant's residence and workplace were measured for 16 carbonyl compounds during summer-fall 1997 as a part of the Air Pollution Exposure Distributions within Adult Urban Populations in Europe (EXPOLIS) study in Helsinki, Finland. When formaldehyde and acetaldehyde were excluded, geometric mean ambient air concentrations outside each participant's residence were less than 1 ppb for all target compounds. Geometric mean residential indoor concentrations of carbonyls were systematically higher than geometric mean personal exposures and indoor workplace concentrations. Additionally, residential indoor/outdoor ratios indicated substantial indoor sources for most target compounds. Carbonyls in residential indoor air correlated significantly, suggesting similar mechanisms of entry into indoor environments. Overall, this study demonstrated the important role of non-traffic-related emissions in the personal exposures of participants in Helsinki and that comprehensive apportionment of population risk to air toxics should include exposure concentrations derived from product emissions and chemical formation in indoor air. PMID- 12774990 TI - Tradescantia pallida cv. purpurea boom in the characterization of air pollution by accumulation of trace elements. AB - Tradescantia pallida cv. purpurea, a plant species widely employed for ornamentation in Brazil, has been successfully used for monitoring the genotoxicity of various agents by the micronucleus assay. To amplify knowledge about its suitability as a bioindicator species, its capacity for accumulating trace elements from urban air pollution was evaluated. T. pallida was rooted using standardized soil, and the vases were distributed in two highly polluted sites of the urban area of Sao Paulo, Brazil (Cerqueira Cesar and Congonhas districts), and in one unpolluted control site situated approximately 50 km from downtown Sao Paulo (in Caucaia do Alto). Approximately six months after exposure to pollutants, adult leaves of this plant were collected monthly for 12 months. The leaves were washed with deionized water, dried, and ground for analyses. Characterization of element levels was carried out by neutron activation analysis. Powdered samples and standards were irradiated at the IEA-R1 nuclear reactor for short and long periods, and concentrations of As, Ba, Br, Ca, Ce, Cl, Cr, Co, Fe, K, La, Mn, Na, Rb, Sb, Sc, Sr, Th, and Zn were determined. Analysis of variance applied to the results indicated that samples from polluted sites present the highest concentrations of Ba, Ce, Cr, Co, Fe, La, Sb, and Sc (p < 0.05). Discriminant analysis revealed that it was possible to distinguish the two polluted areas with a precision of 97.5%, based on the amount of pollutant elements measured in the plants at each site. The results indicated the potential use of T. pallida as an accumulator plant for air pollution biomonitoring. PMID- 12774991 TI - Microwave plasma conversion of volatile organic compounds. AB - A microwave-induced, steam/Ar/O2, plasma "torch" was operated at atmospheric pressure to determine the feasibility of destroying volatile organic compounds (VOCs) of concern. The plasma process can be coupled with adsorbent technology by providing steam as the fluid carrier for desorbing the VOCs from an adsorbent. Hence, N2 can be excluded by using a relatively inexpensive carrier gas, and thermal formation of oxides of nitrogen (NOx) is avoided in the plasma. The objectives of the study were to evaluate the technical feasibility of destroying VOCs from gas streams by using a commercially available microwave plasma torch and to examine whether significant byproducts were produced. Trichloroethene (TCE) and toluene (TOL) were added as representative VOCs of interest to a flow that contained Ar as a carrier gas in addition to O2 and steam. The O2 was necessary to ensure that undesirable byproducts were not formed in the process. Microwave power applied at 500-600 W was found to be sufficient to achieve the destruction of the test compounds, down to the detection limits of the gas chromatograph that was used in the analysis. Samples of the postmicrowave gases were collected on sorbent tubes for the analysis of dioxins and other byproducts. No hazardous byproducts were detected when sufficient O2 was added to the flow. The destruction efficiency at a fixed microwave power improved with the addition of steam to the flow that passed through the torch. PMID- 12774992 TI - Application of the tracer-aerosol gradient interpretive technique to sulfur attribution for the big bend regional aerosol and visibility observational study. AB - A simple data analysis method called the Tracer-Aerosol Gradient Interpretive Technique (TAGIT) is used to attribute particulate S and SO2 at Big Bend National Park in Texas and nearby areas to local and regional sources. Particulate S at Big Bend is of concern because of its effects on atmospheric visibility. The analysis used particulate S, SO2, and perfluorocarbon tracer data from six 6-hr sampling sites in and near Big Bend National Park. The data were collected in support of the Big Bend Regional Aerosol and Visibility Observational (BRAVO) Study; the field portion was conducted from July through October 1999. Perfluorocarbon tracer was released continuously from a tower at Eagle Pass, TX, approximately 25 km northeast of two large coal-fired power plants (Carbon I and II) in Coahuila, Mexico, and approximately 270 km east-southeast of Big Bend National Park. The perfluorocarbon tracer did not properly represent the location of the emissions from the Carbon power plants for individual 6-hr sampling periods and attributed only 3% of the particulate S and 27% of the SO2 at the 6 hr sites in and near Big Bend to sources represented by the tracer. An alternative approach using SO2 to tag "local" sources such as the Carbon plants attributed 10% of the particulate S and 75% of the SO2 at the 6-hr sites to local sources. Based on these two approaches, most of the regional (65-86%) and a small fraction (19-31%) of the local SO2 was converted to particulate S. The analysis implies that substantial reductions in particulate S at Big Bend National Park cannot be achieved by only reducing emissions from the Carbon power plants; reduction of emissions from many sources over a regional area would be necessary. PMID- 12774993 TI - Adjusting tapered element oscillating microbalance data for comparison with Federal Reference Method PM2.5 measurements in region 5. AB - Continuous monitoring of particulate matter (PM) with a diameter less than 2.5 microm (PM2.5) is quickly gaining acceptance as an alternative means of measuring fine PM in the United States. For this project, data were taken from all monitoring sites within Region 5 that used the tapered element oscillating microbalance (TEOM) for PM2.5 and had a collocated Federal Reference Method (FRM) monitor. Scatter plots of TEOM versus FRM show that for a significant fraction of the observations, an independent factor causes the TEOM to underestimate the FRM value. This underestimation appears to increase as temperature decreases. For this analysis, a linear relationship was fit to the TEOM versus FRM data, allowing a break or knot in the relationship, modeled as a change of slope, at a site-specific temperature. To test whether the models are adequate for adjusting future measurements, models were also developed using the first year of data only, and the remaining observations were used to test the durability of the relationships. For all but one monitor in Minnesota, the models developed for each site had consistently high R2s, were predictive of future measurements, and could be used to derive "FRM-like" results from the TEOM measurements. The temperature knots fitted by the model for individual sites ranged from 12.9 to 20.6 degrees C. Data from all six sites in the state of Michigan were also combined to determine if a single model could be developed for the entire state. While the single model for the state of Michigan worked reasonably well, some of the predicted concentrations at individual sites were systematically underestimating the observed concentrations on more polluted days. The same conclusion was drawn for a Region 5-wide model. This approach was also found to work very well for six individual TEOM monitors in New York State. PMID- 12774994 TI - Comparisons of statistical characteristic of air pollutants in Taiwan by frequency distribution. AB - The lognormal, Weibull, and type V Pearson distributions were selected to fit the concentration frequency distributions of particulate matter with an aerodynamic diameter of < or = 10 microm (PM10) and SO2 in the Taiwan area. Air quality data from three stations, Hsin-Chu, Shalu, and Gain-Jin, were fitted with three distributions and compared with the measured data. The parameters of unimodal and bimodal fitted distributions were obtained by the methods of maximum likelihood and nonlinear least squares, respectively. Moreover, the root mean square error (RMSE), index of agreement (d), and Kolmogorov-Smirnov (K-S) test were used as criteria to judge the goodness-of-fit of these three distributions. These results show that the frequency distributions of PM10 concentration at the Hsin-Chu and Shalu stations are unimodal, but the distribution at Gain-Jin is bimodal. The distribution type of PM10 concentration varied greatly in different areas and could be influenced by local meteorological conditions. For SO2 concentration distribution, the distributions were all unimodal. The results also show that the lognormal distribution is the more appropriate to represent the PM10 distribution, while the Weibull and lognormal distributions are more suitable to represent the SO2 distribution. Moreover, the days exceeding the air quality standard (AQS) (PM10 > 125 microg/ m3) for the Hsin-Chu, Shalu, and Gain-Jin stations in the coming year are successfully predicted by the theoretic distributions. PMID- 12774995 TI - PM2.5 assessment in 21 European study centers of ECRHS II: Method and first winter results. AB - The follow-up of a cohort of adults from 29 European centers of the former European Community Respiratory Health Survey (ECRHS) I (1989-1992) will examine the long-term effects of exposure to ambient air pollution on the incidence, course, and prognosis of respiratory diseases, in particular asthma and decline in lung function. The purpose of this article is to describe the methodology and the European-wide quality control program for the collection of particles with 50% cut-off size of 2.5 microm aerodynamic diameter (PM2.5) in the ECRHS II and to present the PM2.5 results from the winter period 2000-2001. Because PM2.5 is not routinely monitored in Europe, we measured PM2.5 mass concentrations in 21 participating centers to estimate background exposure in these cities. A standardized protocol was developed using identical equipment in each center (U.S. Environmental Protection Agency Well Impactor Ninety-Six [WINS] and PQ167 from BGI, Inc.). Filters were weighed in a single central laboratory. Sampling was conducted for 7 days per month for a year. Winter mean PM2.5 mass concentrations (November 2000-February 2001) varied substantially, with Iceland reporting the lowest value (5 microg/m3) and northern Italy the highest (69 microg/m3). A standardized procedure appropriate for PM2.5 exposure assessment in a multicenter study was developed. We expect ECRHS II to have sufficient variation in exposure to assess long-term effects of air pollution in this cohort. Any bias caused by variation in the characteristics of the chosen monitoring location (e.g., proximity to traffic sources) will be addressed in later analyses. Given the homogenous spatial distribution of PM2.5, however, concentrations measured near traffic are not expected to differ substantially from those measured at urban background sites. PMID- 12774996 TI - A methodology to estimate odors around landfill sites: the use of methane as an odor index and its utility in landfill siting. AB - The estimation of odor production and dispersion from landfill sites is a very complicated task because of the different chemical species that exist in biogas. To site a new landfill, it is necessary to know the distance that odors can travel around the landfill under atmospheric conditions that increase the concentration of pollutants. Although CH4 is an odorless gas, it can be used as an index to determine the dispersion of low-reactivity odorous species around a landfill site. Methane production rates, estimated by biogas production models, were used by an air dispersion model to determine the spatial distribution of CH4 around landfill sites. By utilizing dispersion models under extreme atmospheric conditions, a maximum CH4 concentration around the landfills was determined. Based on the ratio between CH4 and odorous chemical species, the spatial distribution of the concentration of an odorous species was determined for those species with low reactivity in the atmosphere. For odorous species with high reactivity in the atmosphere, a dispersion-reaction model must be used. In this way, the acceptable distance between new landfills and residential areas can be determined. The proposed methodology could be used as a design tool for those who are interested in landfill siting. PMID- 12774997 TI - Limb length inequality: clinical implications for assessment and intervention. AB - The purpose of this paper is to review relevant literature concerning limb length inequalities in adults and to make recommendations for assessment and intervention based on the literature and our own clinical experience. Literature searches were conducted in the MEDLINE, PubMed, and CINAHL databases. Limb length inequality and common classification criteria are defined and etiological factors are presented. Common methods of detecting limb length inequality include direct (tape measure methods), indirect (pelvic leveling), and radiological techniques. Interventions include shoe inserts or external shoe lift therapy for mild cases. Surgery may be appropriate in severe cases. Little agreement exists regarding the prevalence of limb length inequality, the degree of limb length inequality that is considered clinically significant, and the reliability and validity of assessment methods. Based on correlational studies, the relationship between limb length inequality and orthopaedic pathologies is questionable. Stronger support for the link between low back pain (LBP) and limb length inequality is provided by intervention studies. Methods involving palpation of pelvic landmarks with block correction have the most support for clinical assessment of limb length inequality. Standing radiographs are suggested when clinical assessment methods are unsatisfactory. Clinicians should exercise caution when undertaking intervention strategies for limb length inequality of less than 5 mm when limb length inequality has been identified with clinical techniques. Recommendations are provided regarding intervention strategies. PMID- 12774998 TI - Reliability of classifications derived from Cyriax's resisted testing in subjects with painful shoulders and knees. AB - STUDY DESIGN: Intrarater and interrater reliability. OBJECTIVES: Examine intrarater and interrater reliability of the resisted-testing component of Cyriax's selective tension testing for patients with painful shoulders and knees. BACKGROUND: Clinicians make diagnostic and intervention decisions about lesions in contractile tissues based on resisted testing. Diagnostic and intervention decisions require reliable data gathering, especially when more than 1 physical therapist manages a patient. No studies have examined agreement of the results of the resisted tests used in selective tension testing, either within or between physical therapists, in subjects having pathology. METHODS AND MEASURES: Subjects with pain in 1 knee (18 male, 22 female; mean age +/- SD = 31.8 +/- 9.5 years) or shoulder (21 male, 25 female; mean age +/- SD = 34.3 +/- 12.9 years) were examined twice. Referring diagnoses included ligament injuries, overuse syndromes, joint instability, and postsurgical symptoms, with some subjects seeking initial diagnosis. Two physical therapists used standardized positions to evaluate 2 knee motions or 6 shoulder and elbow motions. Evaluators applied maximal isometric manual resistance and rated the contraction as strong or weak while subjects identified the presence or absence of pain during the contraction. Evaluators did not interview the subjects and were masked to previous test results. Analyses included percentage of agreement, kappa coefficients, confidence intervals, and maximum kappa coefficients. RESULTS: Intrarater kappa coefficients ranged from 0.44 to 0.82 and interrater coefficients ranged from 0.00 to 0.46. The small number of subjects who were classified as weak affected the kappa coefficients. In the intrarater condition, evaluators averaged 91% of maximum kappa for the knee and 66.5% for the shoulder. In the interrater condition, they averaged 60.4% of the maximum kappa for both the knee and the shoulder. CONCLUSIONS: Based on 2 physical therapist evaluators with previous education in the selective tension system and an additional 6 hours of formal training on the methods, intrarater reliability of resisted tests was generally acceptable for the knee but not for the shoulder. Interrater reliability of these tests, however, was generally not acceptable. Results were limited by subjects who were younger and had mostly chronic conditions that were mildly to moderately severe and by the small subject samples in the analyses. Reliability might be improved by more intensive training of the evaluators and by standardizing the magnitude of the applied resistance and stabilization of the subjects. PMID- 12774999 TI - Surface electromyographic analysis of exercises for the trapezius and serratus anterior muscles. AB - STUDY DESIGN: This study used a prospective, single-group repeated-measures design to analyze differences between the electromyographic (EMG) amplitudes produced by exercises for the trapezius and serratus anterior muscles. OBJECTIVE: To identify high-intensity exercises that elicit the greatest level of EMG activity in the trapezius and serratus anterior muscles. BACKGROUND: The trapezius and serratus anterior muscles are considered to be the only upward rotators of the scapula and are important for normal shoulder function. Electromyographic studies have been performed for these muscles during active and low-intensity exercises, but they have not been analyzed during high intensity exercises. METHODS AND MEASURES: Surface electrodes recorded EMG activity of the upper, middle, and lower trapezius and serratus anterior muscles during 10 exercises in 30 healthy subjects. RESULTS: The unilateral shoulder shrug exercise was found to produce the greatest EMG activity in the upper trapezius. For the middle trapezius, the greatest EMG amplitudes were generated with 2 exercises: shoulder horizontal extension with external rotation and the overhead arm raise in line with the lower trapezius muscle in the prone position. The arm raise overhead exercise in the prone position produced the maximum EMG activity in the lower trapezius. The serratus anterior was activated maximally with exercises requiring a great amount of upward rotation of the scapula. The exercises were shoulder abduction in the plane of the scapula above 120 degrees and a diagonal exercise with a combination of shoulder flexion, horizontal flexion, and external rotation. CONCLUSION: This study identified exercises that maximally activate the trapezius and serratus anterior muscles. This information may be helpful for clinicians in developing exercise programs for these muscles. PMID- 12775000 TI - Clinical decision making in the identification of patients likely to benefit from spinal manipulation: a traditional versus an evidence-based approach. PMID- 12775001 TI - Self-reported giving-way episode during a stepping-down task: case report of a subject with an ACL-deficient knee. AB - STUDY DESIGN: Case report. OBJECTIVE: To describe the knee kinematics and moments of a giving-way trial of a subject with an anterior-cruciate-ligament- (ACL) deficient knee relative to his non-giving-way trials and to healthy subjects during a step-down task. BACKGROUND: Episodes of giving way are believed to damage joint structures, therefore treatments aim to prevent giving-way episodes, yet few studies document giving-way events. METHODS: The giving-way trial experienced by a 32-year-old male subject with ACL deficiency during a step-down task was compared to his non-giving-way trials (n = 5) and data from healthy subjects (n = 20). Position data collected at 60 Hz were combined with anthropometric data and ground reaction force data collected at 300 Hz to estimate knee displacement and 3-dimensional angles and net joint moments. RESULTS: The knee joint displacement was higher during the giving-way trial: from 4% to 32% of stance, reaching 9.0 mm at 18% of stance as compared to 1.6 +/- 0.7 mm for the non-giving-way trials. After 4% of stance, the knee flexion angle of the giving-way trial was 6.6 degrees higher than the non-giving-way trials and was associated with a higher knee extension moment. The knee frontal plane moment was near neutral during early stance of the giving-way trial in contrast to the non-giving way and healthy subjects which demonstrated a knee abduction moment. CONCLUSIONS: The response of this subject to the giving-way event suggests that higher knee flexion angles may enhance knee stability and, in reaction to the giving-way event, that knee extension moment may increase. PMID- 12775002 TI - Evaluation of protocols for field decontamination before bacterial sampling of root canals for contemporary microbiology techniques. AB - The effectiveness of sodium hypochlorite (NaOCl) (2.5%) or iodine (10%) for decontamination of the operation field (tooth, rubber dam, and gasket [Oraseal]) was compared by using bacterial cultivation. In addition, the final samples were also assessed for bacteria by using polymerase chain reaction. Teeth (n = 63) receiving root canal treatment were polished with pumice, isolated with rubber dam, and their margins sealed with Oraseal. The operation field was disinfected with hydrogen peroxide (30%), followed by iodine (n = 31) or NaOCl (n = 32), before and after access cavity preparation. The operation field was sampled before and after each decontamination, giving four samples per field. After the final decontamination, there was no significant difference (p = 0.602, 0.113, 0.204) in recovery of cultivable bacteria from various sites in either group. However, bacterial DNA could be detected significantly (p = 0.010) more frequently from the tooth surfaces after iodine (45%) compared with NaOCl (13%) decontamination, although on the rubber dam or Oraseal surfaces there was no difference. Root canal sampling for polymerase chain reaction might be better preceded by NaOCl decontamination than by iodine, based on the findings. PMID- 12775003 TI - Cytomegalovirus and Epstein-Barr virus active infection in periapical lesions of teeth with intact crowns. AB - Herpesviruses seem to play an important role in the pathogenesis of aggressive periodontitis and may also contribute to periapical pathosis. This study determined the presence of human cytomegalovirus, Epstein-Barr virus, and herpes simplex virus productive infection in five symptomatic periapical lesions of teeth having intact crowns and calcified necrotic pulps. Periapical samples were collected in conjunction with periapical surgery and kept frozen until virological examination. Reverse transcription-polymerase chain reaction was used in herpesviral identification. RNA was isolated from periapical tissue by a guanidinium isothiocyanate-acid phenol procedure. cDNAs were generated from highly conserved regions of the test viruses using a preamplification kit. Sensitivity and validity of the PCR-primers were determined according to established methods. Amplification products were identified using gel electrophoresis. Human cytomegalovirus and Epstein-Barr virus dual transcription was detected in all five periapical lesions studied. Herpes simplex virus transcript was not identified in any lesion. The present data suggest that human cytomegalovirus or Epstein-Barr virus activation participate in the pathogenesis of symptomatic periapical lesions. We hypothesize that periapical active herpesvirus infection impairs local defenses, thereby inducing overgrowth of endodontopathic bacteria and the clinical flare-up of inflammation. PMID- 12775004 TI - Histological and scanning electron microscopy assessment of various vital pulp therapy materials. AB - Pulp capping and pulpotomy procedures were performed on 15 male mongrel dogs. Three materials were used: calcium hydroxide, acid-etched dentin bonding, and mineral trioxide aggregate. Six of the animals were killed at 50 days and nine were killed at 150 days. Samples from 11 dogs were used for histological evaluation, and the remaining dog samples were used for scanning electron microscopy evaluation. Each slide was graded histologically according to previously published criteria. Scanning electron microscopy analysis was performed, and the weight percentage of elements found in the dentin of a nontreated tooth versus the bridge formed in the exposed specimen was established. By evaluating pulp inflammation in vital pulp-therapy treatments, it was found that mineral trioxide aggregate was not significantly different from the untreated control group, both in pulp-capping procedures at 50 days (p = 0.357) or 150 days (p = 0.198) and pulpotomy procedures at 50 days (p = 0.357) or 150 days (p = 0.198). Moreover, histologically mineral trioxide aggregate was a considerably better material than calcium hydroxide or acid-etched dentin bonding in maintaining the integrity of the pulp. PMID- 12775005 TI - Quantitative analysis of the solubilizing action of MTAD, sodium hypochlorite, and EDTA on bovine pulp and dentin. AB - Necrotic bovine pulp and dentin were used in this study as model tissues to represent the organic and inorganic components of the smear layer present in instrumented root canals. The capabilities of endodontic irrigants to dissolve pulverized forms of these tissues were compared. Lyophilized tissue samples were mixed for 2 h at 37 degrees C with MTAD, three concentrations of sodium hypochlorite (NaOCl), 17% EDTA, or isotonic saline. Undissolved tissues were rinsed with water and lyophilized. The change in tissue weight after exposure to an irrigant was measured to quantify solubilization. The results showed that various concentrations of NaOCl removed organic components of pulp and dentin effectively. As pulp solubilizers 5.25% and 2.60% NaOCl were equal (>90%), and 5.25% NaOCl was capable of dissolving virtually the entire organic component of dentin. EDTA was capable of solubilizing inorganic material in dentin and organic material in pulp and likely also in dentin. It dissolved >70% of the dentin and >51% of the pulp. The solubilizing effects of MTAD on pulp and dentin were somewhat similar to those of EDTA. The major difference between the actions of these solutions was a high binding affinity of doxycycline present in MTAD for the dentin. PMID- 12775006 TI - Efficacy of calcium hydroxide: chlorhexidine paste as an intracanal medication in bovine dentin. AB - The purpose of this study was to evaluate the antibacterial efficacy of an intracanal medication composed of calcium hydroxide with 2% chlorhexidine. Dentin from 24 bovine incisors was used. The incisors were made into standardized cylindrical segments of dentin and infected with Enterococcus faecalis. They were then treated with an intracanal paste composed of calcium hydroxide and sterile water or an intracanal paste composed of calcium hydroxide and 2% chlorhexidine for 1 week. Dentin shavings were collected, suspended in solution, and spread on brain-heart infusion agar. After incubation, colony-forming units were enumerated. The amount of bacteria per mg of dentin was determined. The calcium hydroxide paste with 2% chlorhexidine was significantly more effective at killing E. faecalis in the dentinal tubules than calcium hydroxide with water. PMID- 12775008 TI - Cleaning efficiency of nickel-titanium GT and .04 rotary files when used in a torque-controlled rotary handpiece. AB - This study determined if the cleaning efficiency of nickel-titanium rotary files in an endodontic electric handpiece using a no-torque control setting was superior to that obtained when using the torque-control feature. Fifty extracted human anterior teeth with straight canals were divided into two groups of 20 and two control groups of 5. Canals were instrumented with GT and .04 ProFile nickel titanium files until a size 35 advanced to working length. Samples were sectioned and the apical 6 mm of the canal was photographed (x20) and projected onto a 3- x 4-foot grid with squares measuring 0.5 inches each. Total debris was the percentage of the number of squares containing debris versus the total number of squares. The teeth in the torque-controlled group showed an average of 24.99% debris versus 15.55% for the teeth in the no-torque group. The difference was not statistically significant; therefore, no difference can be said to exist between the two torque settings in terms of cleaning efficiency. PMID- 12775007 TI - Additive antimicrobial activity of calcium hydroxide and chlorhexidine on common endodontic bacterial pathogens. AB - It is an established procedure to use calcium hydroxide [Ca(OH)2] as a therapeutic component to achieve sterilization of infected root canals. Unfortunately, some bacterial species are relatively resistant to Ca(OH)2 and are therefore associated with treatment failures. The objective of this study was to identify combinations of substances that improve antimicrobial activity. Ca(OH)2 was used as a suspension or as a component of gutta-percha points. Efficacy of killing was compared for (a) Ca(OH)2 suspension alone; (b) zinc oxide (ZnO) points alone; (c) Ca(OH)2 suspension combined with ZnO points; (d) Ca(OH)2 suspension combined with ZnO/chlorhexidine points; and (e) Ca(OH)2 points combined with ZnO/chlorhexidine points. Common endodontopathogenic bacterial species (Enterococcus faecalis, Fusobacterium nucleatum, Peptostreptococcus micros, Porphyromonas gingivalis, Streptococcus intermedius) in pure cultures served as target organisms. In the in vitro assay, the antibacterial substances and approximately 10(7) colony forming units of test bacteria were co-incubated in diluted human serum under growth conditions appropriate for the single bacterial species. Samples were taken on days 0, 1, 2, 4, 7, and 14 of incubation and viable counts determined. Gram-negative bacteria were sufficiently killed by Ca(OH)2 alone. Combinations with other agents did not improve the killing kinetics. For the inactivation of the Gram-positive bacteria P. micros and S. intermedius, a combination of Ca(OH)2 and ZnO/chlorhexidine, killed the bacteria faster than Ca(OH)2 alone. For E. faecalis, combining Ca(OH)2 suspension with ZnO/chlorhexidine points lead to a faster decrease in the overall number of viable bacteria, but no test conditions lead to the complete loss of culture viability. The results of this study support the use of a combination of Ca(OH)2 and chlorhexidine for faster eradication of P. micros and S. intermedius from infected root canals. PMID- 12775009 TI - Conventional versus storage phosphor-plate digital images to visualize the root canal system contrasted with a radiopaque medium. AB - The pulp tissue was removed from 20 mandibular first molar teeth using 2.5% NaOCl irrigation and hand files. The dried canals were infused with radiopaque contrast medium. Standardized conventional and Digora digital images were obtained of each tooth positioned in a dried mandible at 0- and 30-degree horizontal angulations. Three evaluators rated the image clarity of the 0- and 30-degree original, enhanced, three-dimensional, zoom, and reverse digital image modes as superior, equal, or inferior to corresponding 0- and 30-degree conventional radiographs. The ratings were compared using the Wilcoxon signed rank test. The original, three-dimensional, zoom, or reverse digital images were inferior to the conventional radiographs for clarity of canal anatomy. The enhanced digital images were not always inferior to the conventional radiographs and were the only images superior to the original digital images. Overall, evaluators rated the image clarity of root canal anatomy on conventional radiographs better than on Digora images. However, factors in the experimental design may have contributed to this result. PMID- 12775010 TI - Taurodontism: an endodontic challenge. Report of a case. AB - Taurodontism is a morphoanatomical change in the shape of a tooth, which usually occurs in multirooted teeth. An enlarged body and pulp chamber, as well as apical displacement of the pulpal floor, are characteristic features. Endodontic treatment of a taurodont tooth is challenging, because it requires special care in handling and identifying the number of root canals. A case of endodontic treatment of a maxillary first molar with taurodontism is presented. PMID- 12775011 TI - Identifying multiplanar root canal curvatures using stainless-steel instruments. AB - Information obtained from careful observation of stainless-steel hand file deformation can be a critical step in efficient root canal space debridement. Secondary and tertiary canal curvatures that occur most often in the proximal view have been reported in the literature. Except in teeth that are clinically rotated, there is currently no method for observing the proximal view of root canal systems. This article presents a simple clinical procedure for detecting the three-dimensional anatomy of root canal spaces. We refer to the procedure as file molding. PMID- 12775012 TI - S-1 in gastric cancer: a comprehensive review. AB - The current basic and clinical studies of S-1 (TS-1) were reviewed. S-1 is a novel oral dihydropyrimidine dehydrogenase (DPD) inhibitory fluoropyrimidine (DIF) based on a biochemical modulation of 5-fluorouracil (5-FU); S-1 contains tegafur (FF) and two types of enzyme inhibitor, 5-chloro-2,4-dihydroxypyridine (CDHP) and potassium oxonate (Oxo) in a molar ratio of 1:0.4:1. In pharmacokinetic studies, S-1 showed high 5-FU concentration in blood for long periods of time. In a combined analysis of two pivotal late phase II studies in gastric cancer, the overall response rate was 44.6% (45/101), and median survival time and 1-year survival rate were 244 days and 37%, respectively. A postmarketing survey was conducted, and in the interim analysis, tolerability and safety profiles were shown in 3294 patients with gastric cancer. The oral dose form and low incidence of adverse reactions permit treatment on an outpatient basis. To evaluate the survival benefit of S-1 in advanced gastric cancer, a phase III study of S-1 vs 5-FU vs cisplatin (CDDP) plus irinotecan (CPT-11) has been conducted. The effect of S-1 in adjuvant chemotherapy is also promising. Currently, a phase III study of surgery alone vs S-1 in patients with curative resection of gastric cancer is in progress. Further therapeutic benefits are expected to be gained by combining S-1 with other chemotherapeutic agents. Several preliminary results of combination phase I/II studies of S-1 with CDDP or CPT-11 have recently been obtained, and phase II studies are in progress. Thus, S 1 is currently the first candidate as the standard anticancer drug for gastric cancer. Further evaluations by well-controlled clinical trials are still needed. PMID- 12775013 TI - Current topics of S-1 at the 74th Japanese Gastric Cancer Congress. AB - S-1 (TS-1)-related studies presented at the 74th Japanese Gastric Cancer Congress are reviewed. Of the papers presented at this conference, 35 were related to S-1. In the panel discussion on the therapeutic significance of oral fluoropyrimidines in gastric cancer, 9 papers were related to S-1 (sensitivities to oral fluoropyrimidines, 2 papers; clinical results of treatment with S-1, 5 papers; and combination therapy with S-1, 2 papers). In the general presentations, there were 26 papers on S-1 related-subjects (clinical studies or clinical practice of S-1, 12 papers; case reports, 3 papers; basic studies on animal models of peritoneal metastasis, 2 papers; and combination therapy with S-1, 9 papers). Several studies showed that S-1 was basically as effective against tumors in postmarketing surveillance in clinical practice as in phase II studies at the time of its development, including a report of a patient with complete response to S-1. Some reports suggested the possibility of using S-1 in neoadjuvant chemotherapy and postoperative adjuvant chemotherapy. The usefulness of S-1 in combination chemotherapy was also suggested in several reports. These results indicate that S-1 is a key drug that can be used in first-line treatment of gastric cancer. It will be necessary to accumulate evidence based on data from clinical trials and clinical applications in the future. PMID- 12775014 TI - Experimental study to evaluate the usefulness of S-1 in a model of peritoneal dissemination of gastric cancer. AB - BACKGROUND: Favorable results have been reported for the novel oral anticancer agent S-1 (TS-1) in clinical studies of advanced gastric cancer with peritoneal dissemination. In the present study we assessed its pharmacokinetics, inhibitory effects, and effect on survival time in an animal model. METHODS: A model of peritoneal dissemination was created by intraperitoneally implanting 4-week-old female BALBc nu/nu mice with the human gastric cancer cell line MKN-45 after transfection with a fluorescent protein-expressing vector. Pharmacokinetics were investigated by measuring intratumor, peritoneal lining, and blood concentrations after the administration of S-1 and fluorouracil (5-FU). The effect of S-1 on survival time was also assessed, by administration once daily to seven animals per group, starting on day 7 after implantation, and survival time was compared with that of an untreated control group. The inhibitory effect of S-1 on peritoneal dissemination was evaluated by killing mice at the start of administration, and 1 and 3 weeks after the start of administration, and examining them for the presence of peritoneal dissemination under a fluorescence stereomicroscope. RESULTS: Maintenance of high 5-FU concentrations in the intraperitoneal tumors was confirmed in the S-1 group, and survival time was prolonged without any decrease in oral food intake or body weight. CONCLUSION: Assessment in a model of peritoneal dissemination of gastric cancer showed that the novel oral anticancer agent S-1 was effective against peritoneal dissemination, and that it improved the survival rate. PMID- 12775015 TI - Efficacy and safety profile of S-1 in patients with metastatic gastric cancer in clinical practice: results from a post-marketing survey. AB - BACKGROUND: S-1(TS-1), a novel oral fluoropyrimidine, has been commercially available for gastric cancer in Japan. A nationwide post-marketing survey for safety was carried out after its approval. The aim of this analysis was to evaluate the efficacy and safety profile of this agent in clinical practice for patients with advanced gastric cancer registered in the postmarketing survey from our institution. METHODS: Between April 1999 and April 2000, a total of 51 chemo naive patients were registered in the survey from the National Cancer Center Hospital East. S-1 was administered at 80 mg/m2/day for 4 weeks, followed by a 2 week rest, repeated every 6 weeks until disease progression, unacceptable toxicity, or the patient's refusal. RESULTS: Of the 51 patients, 41 (80%) fulfilled the criteria of the guidelines determined by the company as appropriate patients for the drug administration. The median number of treatment courses was five. Toxicities were generally mild: grade 3 or 4 toxicities were seen in 10% or fewer patients, and no treatment-related deaths occurred. In the 47 patients with evaluable lesions, there were 2 complete responses and 18 partial responses, with a response rate of 43%. With a minimum follow-up of 2 years, median survival time and 2-year survival were 11.1 months and 33%, respectively. The majority of the 17 2-year survivors had diffuse-type histology and peritoneal metastasis and achieved an objective response. CONCLUSION: S-1 appears to be safe and highly active, with favorable longterm survival in patients with metastatic gastric cancer, particularly in those with diffuse-type histology and peritoneal metastasis. PMID- 12775016 TI - Longterm control of advanced and recurrent gastric cancer (ARGC) by S-1. AB - BACKGROUND: An oral tegafur compound, S-1 (TS-1), was developed to potentiate antitumor activity and to reduce gastrointestinal toxicities for patients with gastric cancer. It has achieved a high response rate against advanced and recurrent gastric cancer (ARGC) in Japan; however, the efficacy and adverse reactions of longterm administration of S-1 remain to be elucidated. METHODS: Sixty-nine patients with ARGC treated with S-1 were studied; 58 patients had measurable lesions, while 11 patients did not. S-1 was orally administered at doses of between 40 and 60 mg/body twice daily for 28 days, followed by 14 days' rest, as one course. RESULTS: The overall response rate was 38% (complete response [CR], 2/58; partial response [PR], 25/58; stable disease [SD], 9/58 progressive disease [PD] 23/58). Response rate by target organ was 40% for the primary lesion, 45% for lymph node metastasis, 38% for peritoneal metastasis, and 25% for liver metastasis. When S-1 was administered as second-line chemotherapy (n = 25), the response rate was 36%. Of the 69 patients, 14 received S-1 for more than a year. The median survival time (MST) after S-1 administration in these 14 patients, including 3 patients with stable disease, was 918 days (range, 536 to 1107 days). There were no grade 3 to 4 toxicities in these 14 patients receiving longterm therapy with S-1. CONCLUSION: S-1 therapy was performed with a high response rate, irrespective of the target organ or the presence of prior chemotherapy. Longterm administration of S-1 may benefit patients with ARGC, providing prolonged disease control with acceptable toxicities. PMID- 12775017 TI - S-1 in the treatment of advanced and recurrent gastric cancer: current state and future prospects. AB - BACKGROUND: S-1 (TS-1) is a novel oral anticancer drug. Because of the excellent results of phase II studies, we continued to prescribe S-1 for advanced or recurrent gastric cancer after we participated in the phase I and II studies. METHODS: Twenty-nine patients with advanced or recurrent gastric cancer were treated with S-1. Clinicopathological features, survival, and adverse reactions were analyzed. RESULTS: One course of treatment consisted of 40, 50, or 60 mg/body twice a day for 28 days followed by withdrawal for 2 weeks. The mean number of treatments was 3.6 courses (range, 1-12 courses). The response rate was 37.9% (11 partial responses [PRs] in 29 patients). Although the response rate of patients who did not receive prior chemotherapy was 47.6% (10 PRs in 21 patients), that of patients with prior chemotherapy was 12.5% (1 PR in 8 patients). The median survival time was 14.1 months, and that of patients who responded to treatment was 22.1 months, which was significantly longer than that of nonresponder patients. One-year and 2-year survivals in the 29 patients were 50.2% and 24.3%, respectively. Adverse reactions were noted in 17 of 29 patients, and the most frequent one was leukocytopenia. Only 2 patients experienced grade 3 leukocytopenia and neutrocytopenia. CONCLUSION: Because of the high response rate and low incidence of severe adverse reactions, S-1 is a first-line chemotherapy that can be used for outpatients, especially for patients without prior chemotherapy. As the response rate for patients with prior chemotherapy was low, combined therapy with S-1 is worth evaluating for these patients. PMID- 12775018 TI - A new regimen for S-1 therapy aiming at adverse reaction mitigation and prolonged medication by introducing a 1-week drug-free interval after each 2-week dosing session: efficacy and feasibility in clinical practice. AB - BACKGROUND: The response rate of advanced or recurrent gastric cancer to S-1 (TS 1) is 46.5%, which is higher than the response rate of this type of cancer to any other anticancer agent. However, the incidence of adverse reactions to this drug has also been reported to be as high as 83.2%. According to a postmarketing survey, adverse reactions to this drug begin to appear 2-3 weeks after the start of drug administration. With these findings in mind, we recently devised a new dosing regimen for the drug, by which the drug is administered for 2-week periods separated by 1-week drug-free intervals (the 2-week regimen). The aim of this retrospective study was to evaluate the efficacy and feasibility of the 2-week regimen in comparison with a 4-week dosing regimen with a 2-week interval between sessions (the 4-week regimen) as the historical control. METHODS: The subjects were 27 patients with advanced or recurrent gastric cancer who received S-1 therapy at our center between September 1999 and November 2001. Of these patients, 14 who received the 4-week regimen before January 2001 served as historical controls, and the results in these patients were compared with those of the remaining 13 patients, who received the 2-week regimen after February 2001. Patient backgrounds, adverse reactions, compliance, and efficacy were investigated retrospectively. RESULTS: The incidence of adverse reactions tended to be lower in the 2-week-regimen group (77%) than in the 4-week-regimen group (93%). The percentage of patients who received the drug for 6 months in complete compliance with the dosing schedule, as calculated by the Kaplan-Meier method, was 85% in the 2-week-regimen group and 40% in the 4-week-regimen group. The response rate to the drug was 23% in the 2-week-regimen group and 21% in the 4 week-regimen group. CONCLUSION: These results suggest that this 2-week regimen may mitigate adverse reactions and prolong the medication period. PMID- 12775019 TI - Neoadjuvant chemotherapy with S-1 for scirrhous gastric cancer: a pilot study. AB - We conducted a pilot study using S-1 (TS-1), a novel oral derivative of 5 fluorouracil, as neoadjuvant chemotherapy for potentially resectable scirrhous gastric cancer. The neoadjuvant chemotherapy consisted of two courses (each, 4 week administration and 2-week withdrawal) of S-1 at 100-120 mg/body per day. Five patients were enrolled in this pilot study and underwent resection. The response rate for the neoadjuvant chemotherapy was 60% (three partial response [PR]; two stable disease [SD]). Three of the five patients received curative resection; the other two patients received noncurative resection because of localized peritoneal dissemination and positive results on cytological examination of the abdominal washing. No toxicity of grade 3 or more was exhibited during the two courses of chemotherapy. Pathological examination of the resected specimens revealed a marked reduction in the distribution of viable cancer cells in the stomach in the three patients with PR. In one of these patients, pathological findings suggestive of the possibility of disappearance of the cancer cells in the perigastric and paraaortic lymph nodes were noted. Because of the unexpectedly high response to S-1, we consider that the efficacy of S-1 as neoadjuvant chemotherapy for scirrhous gastric cancer should be verified by phase II and III trials. PMID- 12775020 TI - Clinical efficacy of S-1 combined with cisplatin for advanced gastric cancer. AB - Several chemotherapy regimens used against advanced gastric cancer have been studied extensively over the decades in an attempt to further improve the prognosis of patients. To date, no standard chemotherapeutic regimens have been established worldwide. S-1 (TS-1), a combination of ftorafur and two modulators, gimestat (CDHP) and oxonic acid, in a molar ratio of 1:0.4:1, has been widely used in Japan for the treatment of advanced gastric cancer, and much attention has been paid to attempts to increase its antitumor effect by combining it with other chemotherapeutic drugs. We treated 12 patients with advanced gastric cancer with 80 mg/m2 of S-1 for 21 days and 60 mg/m2 of cisplatin (CDDP) on day 8 every 5 weeks. The treatment was continued until disease progression, unacceptable toxicity, or the patient's refusal. Eight out of 12 evaluable patients achieved a partial response (PR), with a response rate of 66.7%. The incidence of grade 3 or 4 adverse effects, including myelosuppression and gastrointestinal toxicities, was 16.6%. None of the patients treated with this regimen died of adverse effects and none required hospitalization for the toxicity. We conclude that the combination of S-1 and CDDP seems to have a high therapeutic index, enhancing the antitumor effect of S-1 while maintaining modest adverse effects, thus suggesting the possible use of this combination based at the outpatient clinic (apart from a short stay in hospital during the infusion of CDDP with hydration). Further study with a large number of patients may be needed to confirm the combination of S-1 and CDDP to be an appropriate first-line chemotherapy for gastric cancer. PMID- 12775021 TI - Combination chemotherapy of S-1 and low-dose cisplatin for advanced gastric cancer. AB - BACKGROUND: A previous phase II study showed that S-1 (TS-1) was effective for advanced gastric cancers and had mild toxicity. The present study aimed to evaluate the efficacy and feasibility of this novel anticancer drug combined with low-dose cisplatin (CDDP). METHODS: Fifteen patients with unresectable and recurrent gastric cancer were enrolled. S-1 was administered orally twice daily after meals, at a standard dose of 80 mg/m2 per day according to the late phase II trial protocol. One course consisted of 28 days' consecutive administration followed by 14 days' rest. Five or 10 mg CDDP was infused three times each week (days 1, 3, 5) during S-1 administration on hospitalization, and once each week (day 1) at the outpatient clinic. Patients' backgrounds, response rates, response durations, and time to progression were investigated. RESULTS: None of the 15 patients had a complete response and 8 had a partial response. Therefore, the overall response rate was 53% (8/15). For site efficacy, the response rate was 50% (5/10) for the primary lesion, 50% (3/6) for liver metastasis, and 39% (5/13) for lymph node metastasis. The median response duration of the 8 responders was 4 months, and the time to progression was 3.3 months. Adverse reactions appeared in 60% (9/15). The incidence of adverse reactions of grades 3 and 4 was 13% (2/15) and 0%, respectively. As for hematological toxicity, grade 3 leukopenia was observed in 2 patients (2/15), but decreased hemoglobin level and thrombocytopenia did not appear. Although gastrointestinal adverse reactions appeared in 40% (6/15), all reactions were grades 1 and 2. Because of the mild toxicity, most patients were treated at the outpatient clinic. CONCLUSIONS: Combination chemotherapy of S-1 and low-dose CDDP is expected to be a useful chemotherapy for advanced gastric cancer. PMID- 12775022 TI - Development of clinical pathway in S-1 chemotherapy for gastric cancer. AB - We have developed a clinical pathway to enable the safe continuous administration of S-1 (TS-1) in ambulatory care for patients with advanced gastric cancer. The S 1 clinical pathway includes a pathway for clinicians, a pathway for patients, and such assist tools as a medication diary, an explanatory document containing instructions relating to patient compliance, a table of associations between adverse reactions and prodromes, and general principles for dose reduction and withdrawal. These pathways and assist tools will improve the patient's perception of adverse reactions, thereby contributing to early discovery and rapid action against adverse events. The S-1 clinical pathway has been used with ten patients. S-1 administration has been continued in seven patients. In four of the seven patients, continued administration on the occurrence of adverse reactions was made possible by the use of appropriate measures such as drug withdrawal or dose reduction. It was confirmed that the S-1 clinical pathway was a useful tool for cancer chemotherapy in ambulatory care. PMID- 12775023 TI - Dihydropyrimidine dehydrogenase (DPD) activity in gastric cancer tissue and effect of DPD inhibitory fluoropyrimidines. AB - BACKGROUND: Dihydropyrimidine dehydrogenase (DPD) is an enzyme that catabolizes 5 fluorouracil (5-FU). The effect of DPD inhibitory fluoropyrimidines (DIF) is presumably related to DPD activity. We studied the efficacy of DIF (tegafur + uracil UFT], tegafur + gimeracil + osteracil [S-1 (TS-1)]) relative to DPD activity, with other fluoropyrimidines as controls. METHODS: The efficacy of DIF relative to DPD activity was evaluated in 58 gastric cancer patients who received postoperative administration of fluoropyrimidines, consisting of DIF in 42 patients (UFT in 23; S-1 in 19) and non-DIF in 16 patients. RESULTS: In patients with low DPD activity (under 40 U/mg protein), curative potential tended to be lower for DIF than for non-DIF, but the survival rate was the same for both. In patients with high DPD activity (40 U/mg protein or more), such a tendency was not detected. In a comparison between those treated with UFT and those treated with S-1, prognosis was better in the latter group, in spite of their predominance of lower curative potentials of B or C. In 27 patients with measurable lesions, a partial response (PR) or higher response occurred in 33% (5/15) of those with low DPD activity, and in 17% (2/12) of those with high DPD activity. In the patients with low DPD activity, non-DIF induced no change (NC) in 17% (16), and progressive disease (PD) in the rest. UFF induced PD in all 5 patients, while S-1 induced a response rate of 44% (7/16), with NC in 25% (4/16). In the patients with high DPD activity, on the other hand, non-DIF (n = 3) and UFT (n = 3) induced PD in all the patients, while S-1 induced PR in 33% (2/6) and NC or a higher response in 67% (4/6). CONCLUSION: It is recommended to use S-1 rather than UFF in patients with high DPD activity. Measurement of DPD was useful in drug selection. PMID- 12775026 TI - Internalizing the externalities. PMID- 12775027 TI - How charcoal fires heat the world. PMID- 12775025 TI - Future prospects of personalized chemotherapy in gastric cancer patients: results of a prospective randomized pilot study. AB - In the present study, in order to evaluate the feasibility of personalized chemotherapy, a prospective randomized pilot study was performed in 30 advanced or recurrent gastric cancer patients. As we have demonstrated previously, the expressions of mRNA from tumor biopsy samples for seven molecular markers, i.e., dihydropyrimidine dehydrogenase (DPD), glutathione S-transferase (GST)-pi, beta tubulin (tub), O6-methylguanine-DNA methyltransferase (MGMT), multiple drug resistant protein (MRP)-1, NADPH/quinone oxidoreductase (NQO)-1, and cytochrome p450 (P450), were examined by reverse transcription-polymerase chain reaction (RT PCR) analysis and therapy was recommended in a flow chart that depended on the level of expression of these predictive molecular markers. We chose 12 therapeutic plans, including best supportive care (BSC). We treated 15 patients according to the gene expression profiles, and the remaining 15 patients (controls) were treated without recommended regimens, and the therapy was continued after the expression profiles were checked. Interestingly, 11 of 26 lesions (42.3%) responded after treatment given according to gene expression analysis; however, no clinical response was detected in the control group. The prediction of the response, including resistance, was successful in 75.9% by the gene expression profiles. Moreover, the survival of the patients with the recommended treatment was better than that of patients without a recommended protocol. These results indicate that personalized treatment may be beneficial for gastric cancer chemotherapy and further randomized trials should be carried out in the future. PMID- 12775028 TI - Nitrate eyed as endocrine disrupter. PMID- 12775024 TI - Prediction of sensitivity to fluoropyrimidines by metabolic and target enzyme activities in gastric cancer. AB - BACKGROUND: This study was designed to investigate the role of thymidylate synthase (TS), dihydropyrimidine dehydrogenase (DPD), and thymidine phosphorylase (TP) in tumor progression and sensitivity to 5-fluorouracil (5-FU). METHODS: A total of 275 tumor samples from 275 patients with gastric cancer were utilized in this study. TS activity was determined in 130 samples by 5-fluorodeoxyuridine monophosphate binding assay. DPD activity was measured in 140 samples by radioenzymatic assay, and TP protein level was determined in 157 samples by an enzyme-linked immunosorbent assay (ELISA) system. These parameters were compared with several clinicopathologic factors and sensitivity to 5-FU determined by in vitro ATP assay. The antitumor activities of 5-FU, uracil plus tegafur (UFT), and 1M tegafur--0.4 M 5-chloro-2,4-dihydroxypyridine--1 M potassium oxonate (S-1 [TS 1]) were also compared, using three human gastric cancer xenografts in nude mice. RESULTS: There was no correlation between either TS or TP and sensitivity to 5 FU. However, a weak inverse correlation was found between DPD activity and sensitivity to 5-FU. High DPD activity in tumor resulted in poor prognosis, especially in patients who received 5-FU-based adjuvant chemotherapy. Although TP was significantly correlated with depth of tumor invasion and with lymphatic and venous invasions, TP alone had no impact on survival. On the other hand, TS, as well as peritoneal, hepatic, and lymph node metastases, was selected as an independent prognostic factor in gastric cancer. In the animal model, there was no significant difference in antitumor activities among the drugs in a tumor with low DPD activity. However, S-1 showed superior antitumor activity to 5-FU or UFT in tumors with high DPD activity. CONCLUSION: DPD is considered to be a most important predictive factor of 5-FU sensitivity. The use of DPD inhibitory fluoropyrimidines is strongly recommended for tumors with high DPD activity. PMID- 12775029 TI - Common household chemicals affect algae. PMID- 12775030 TI - Why do PBDE levels vary widely? PMID- 12775031 TI - U.K. targets a 60% cut in CO2. PMID- 12775032 TI - Perchlorate regulation faces further delay. PMID- 12775033 TI - Mercury action and inaction. PMID- 12775034 TI - CDC pegs human exposures to chemicals. PMID- 12775035 TI - Microbes manufacture plastic from food waste. PMID- 12775036 TI - Wildlife toxicology revisited. PMID- 12775037 TI - Equilibrium sampling devices. PMID- 12775038 TI - Bacterial formation of tooeleite and mixed arsenic(III) or arsenic(V)-iron(III) gels in the Carnoules acid mine drainage, France. A XANES, XRD, and SEM study. AB - The oxidation of Fe(II) in acid mine drainage (AMD) leads to the precipitation of Fe(III) compounds which may incorporate toxic elements, such as arsenic (As), within their structure or adsorb them at their surface, thus limiting their mobility. The present work provides evidence for spatial and seasonal variations of microbial activity that influence arsenite oxidation and As immobilization in the heavily contaminated AMD from the Carnoules mine, Gard, France ([As III] = 80 to 280 mg x L(-1) in the acidic spring draining the waste-pile). In the first tens of meters of the AMD, the rapid oxidation of Fe(II) leads to the coprecipitation of large amounts of As with Fe(III) in bacterial mats. XRD, XANES, and SEM analyses of sediments and stromatolite samples revealed the unusual formation of As(III)-rich compounds, especially nanocrystalline tooeleite, Fe6(AsO3)4(SO4)(OH)4 x 4H2O, a rare ferric arsenite sulfate oxy hydroxide mineral, together with XRD-amorphous mixed As(III)/As(V)-Fe(III) oxy hydroxide compounds. In the wet season, the suspended sediments of the upstream zone essentially consist of tooeleite associated with am-As(III)-Fe(III) oxy hydroxides, while am-As(V)-Fe(III) oxy-hydroxides, having As:Fe molar ratios as high as 0.6-0.8, dominate in the dry season. Comparing natural and bioassay samples revealed that the formation of As(III)-rich compounds in the wet season may be related to the metabolic activity of bacterial strains able to oxidize Fe(II) but not As(III). One of these strains, having an Acidithiobacillus ferrooxidans genotype, has been isolated from the Carnoules AMD. In contrast, the formation of As(V)-rich compounds in the dry season can be related to both biotic and abiotic oxidation of As(III) to As(V). Some Thiomonas strains isolated from the Carnoules AMD were shown to be able to catalyze the oxidation of As(III) to As(V) in solution. Therefore, they can promote the formation of mixed As(V) Fe(III) oxy-hydroxides, provided enough Fe(II) oxidizes. These results yield a better understanding of natural processes at this site and may help in designing efficient As-removal processes. PMID- 12775039 TI - Effects of three pharmaceutical and personal care products on natural freshwater algal assemblages. AB - Treated wastewaters in the United States contain detectable quantities of surfactants, antibiotics, and other types of antimicrobial chemicals contained in pharmaceutical and personal-care products (PPCPs) that are released into stream ecosystems. The degradation characteristics of many of these chemicals are not yet known, nor are the chemical properties of their byproducts. They also are not currently mandated for removal under the U.S. Clean Water Act. Three representative PPCPs were individually tested in this study using a series of laboratory dilution bioassays: Ciprofloxacin (an antibiotic), Triclosan (an antimicrobial agent), and Tergitol NP 10 (a surfactant), to determine their effects on natural algal communities sampled both upstream and downstream of the Olathe, KS wastewater treatment plant (WWTP). There were no significant treatment effects on algal community growth rates during the exponential phase of growth, but significant differences were observed in the final biomass yields (p < 0.001). All three compounds caused marked shifts in the community structure of suspended and attached algae at both the upstream and downstream sites (p < 0.05). Increasing the concentrations of all three compounds over a 3 orders of magnitude range also caused a consistent decline in final algal genus richness in the bioassays. Our results suggest that these three PPCPs may potentially influence both the structure and the function of algal communities in stream ecosystems receiving WWTP effluents. These changes could result in shifts in both the nutrient processing capacity and the natural food web structure of these streams. PMID- 12775040 TI - Hydroxylated PCBs and other chlorinated phenolic compounds in lake trout (Salvelinus namaycush) blood plasma from the Great Lakes region. AB - Recently, there has been an increase in studies focusing on an emerging class of organic contaminants, hydroxylated PCBs (OH-PCBs) and chlorinated phenolic compounds (CPCs) in the environment, particularly in northern regions of Europe and Canada. Since information for fish from the Great Lakes are scarce, we determined the blood plasma concentrations of OH-PCB congeners, pentachlorophenol (PCP), 2,3,4,5-tetrachlorophenol (TCP), and 4-hydroxyheptachlorostyrene (4-OH HpCS) for lake trout (Savelinus namaycush) collected from two of the Great Lakes, Lake Ontario and Lake Superior, and two regional lakes, Lake Champlain and Lake Opeongo. PCP was the dominant CPC in lake trout (105-658 pg/g of plasma). Detectable concentrations of 2,3,4,5-TCP and 4-OH-HpCS were found in all lake trout (2.6-101 and 0.4-27 pg/g, respectively). Highest concentrations were found in trout from Lake Ontario and Lake Superior. Sixteen OH-PCBs were quantified, with 4-OH-CB187 having the highest concentration in all samples (10-173 pg/g of plasma). Unexpectedly, highly chlorinated OH-PCBs such as 4'-OH-CB199 (mean 21.4 and 74.4 pg/g), 4,4'-diOH-CB202 (18.3 and 27.7 pg/g), and 4'-OH-CB208 (24.5 and 34.7 pg/g) were found in lake trout from Lake Ontario and Lake Superior, respectively. Future studies to delineate the sources and impacts of CPCs in the Great Lakes catchment are needed. PMID- 12775041 TI - Is a universal model of organic acidity possible: comparison of the acid/base properties of dissolved organic carbon in the boreal and temperate zones. AB - The acid/base properties of dissolved organic carbon (DOC) are an important feature of soil and surface waters. Large differences in the acid/base properties of DOC found by different studies might be interpreted as spatial and temporal differences in these properties. Different analytical techniques, however, may explain some of the differences. We used a combination of ion-exchange techniques, titration, and surface water chemistry data to evaluate DOC character from two substantially different areas--the relatively pristine boreal zone of Sweden and the heavily acidified temperate zone of the Czech Republic. We found a significantly higher site density (amount of carboxylic groups per milligram of DOC) for the Swedish sites (10.2 microequiv/mg of DOC +/- 0.6) as compared to the Czech sites (8.8 microequiv/mg of DOC +/- 0.5). This suggests a slightly higher buffering capacity for Swedish DOC. A triprotic model of a type commonly incorporated in biogeochemical models was used for estimating the DOC dissociation properties. For Swedish sites, the following constants were calibrated: pKa1 = 3.04, pKa2 = 4.51, and pKa3 = 6.46, while the constants for Czech sites were pKa1 = 2.5, pKa2 = 4.42, and pKa3 = 6.7. Despite differences in site density values, both models predict very similar dissociation and thus pH buffering by DOC in the environmentally important pH range of 3.5-5.0. This can be incorporated into models to make reliable estimates of the effect of organic acids on pH and buffering in different regions. PMID- 12775042 TI - Retrospective monitoring of organotin compounds in marine biota from 1985 to 1999: results from the German Environmental Specimen Bank. AB - In archived samples from the German Environmental Specimen Bank, organotin compounds including tributyltin (TBT) and triphenyltin (TPT) as well as their degradation products were quantified. Biota samples from North Sea and Baltic Sea areas were analyzed by gas chromatography/atomic emission detection-coupling after extraction and Grignard or ethylborate derivatization. TBT and TPT were detected in nearly all samples. A decrease of TPT contamination was observed in bladder wrack, common mussels, and eelpout muscle tissues in the period 1985 1999. In this period, TPT concentrations in North Sea mussels decreased from 98 to 7 ng/g (as organotin cation concentration in wet tissue). Concentrations of TBT remained relatively constant with 17 +/- 3 ng/g for mussels from a site with nearby marine traffic and 8 +/- 2 ng/g for a more remote area. The results reflect that TBT is still used as a biocide in antifouling paints whereas the use of TPT as a co-toxicant in such preparations had been ceased in the 1980s. The fact that the use of TBT in antifouling paints was banned in 1991 for small boats within the European Community seems not to have resulted in a decrease of TBT levels in marine biota. PMID- 12775043 TI - Reevaluation of air-water exchange fluxes of PCBs in Green Bay and southern Lake Michigan. AB - Air-water exchange of persistent organic pollutants (POPs) such as polychlorinated biphenyls (PCBs) is an import process for the delivery of contaminants to water bodies, as well as for their removal, and is thus a pivotal parameter in the construction of mass balances in aquatic systems. Simultaneous measurements of PCB concentrations in the gas and dissolved phases conducted in Green Bay in 1989 and in southern Lake Michigan in 1994-1995 were used to estimate air-water exchange fluxes. In this work, improved Henry's law constants for PCBs and new mass-transfer rates across the air-water interface were used to update the previous calculations. The new model calculations suggest that the net volatilization flux of PCBs out of Green Bay ranges from +170 to +5300 ng m(-2) day(-1), which is 2-20 times larger than previous estimates. The flux of PCBs in southern Lake Michigan exhibits net volatilization of +0.5 to +230 ng m(-2) day( 1) throughout the study period (May and July 1994, January 1995), whereas previous estimates reported that the net flux was seasonally absorptive. Thus, water-to-air fluxes are more important for the removal of PCBs from both Green Bay and Lake Michigan than previously recognized. PMID- 12775044 TI - Steroid estrogens profiles along river stretches arising from sewage treatment works discharges. AB - Concentrations of estradiol, estrone, and ethinylestradiol were measured in the water column (daily for 28 or 14 days) and in the bed sediment (weekly over the same period) of the River Nene and the River Lea, U.K., upstream and downstream of sewage treatment works (STW). The concentrations of the three steroids in the STW effluents were also measured. Estrone was detected at the highest concentration and in almost all samples from the three STW effluents, concentrations ranging from <0.4 to 12.2 ng/L. Estradiol was also detected frequently (<0.4-4.3 ng/L), but ethinylestradiol was detected infrequently (<0.4 3.4 ng/L). Positive detections were only found for estrone in the sediment, and these seemed to be unrelated to the water column concentrations. Levels of estrone were clearly raised above background levels in the rivers as a result of the STW discharges. Levels of estradiol and ethinylestradiol were too close to their detection limits to assess the STW impact. River water estrone concentration declined downstream at a rate that was in excess of that due to dilution. The most likely cause of this decline is a combination of sorption and biodegradation equivalent to a first-order decay half-life of 2.5 days for the River Nene and 0.5 days for the River Lea. PMID- 12775045 TI - Nuclear magnetic resonance and diffuse-reflectance infrared Fourier transform spectroscopy of biosolids-derived biocolloidal organic matter. AB - We extracted the acid-soluble portion of municipal biosolids, fractionated it by both molecular weight (MW) and hydrophobicity, and used various solid-state nuclear magnetic resonance (NMR) methods and diffuse-reflectance infrared Fourier transform (DRIFT) spectroscopy to characterize the fractions. Spectroscopic characterization of the MW components of the biosolids-derived organic matter fractions revealed the presence of functionally distinct groups of compounds. Quantitative 13C NMR, CH spectral editing, and several two-dimensional NMR experiments show that the high-MW hydrophilic fraction in particular is structurally simple, consisting predominantly of N-acetylated polysaccharides, perhaps derived from bacterial peptidoglycans. In the high-MW hydrophobic fraction, aromatic compounds were present in addition to the N-acetylated polysaccharides. Infrared spectroscopy confirmed that hydrophilic fractions were dominated by carbohydrates and indicated that the lower-MW fractions lacked amide moieties. Complementary interpretations of the DRIFT and NMR spectra improved our knowledge of the components separated by this fractionation scheme, allowing better characterization of biosolids organic matter. Moreover, fractionation based on both MW and hydrophobicity may prove useful in detailed characterization of the structure of biosolids-derived organic matter and other similarly heterogeneous natural organic matter in soils and sediments. PMID- 12775046 TI - PCDD/F, PCB, HxCBz, PAH, and PM emission factors for fireplace and woodstove combustion in the San Francisco Bay region. AB - Emissions from residential fireplace and woodstove appliances burning fuels available from the San Francisco Bay area were sampled for polychlorinated dibenzodioxins and dibenzofurans (PCDDs/Fs), polychlorinated biphenyls (PCBs), hexachlorobenzene (HxCBz), particulate matter (PM), polycyclic aromatic hydrocarbons (PAHs), oxygenated PAHs, and the monosaccharide levoglucosan. Emission factors for these pollutants were determined, the first known characterization of this extent. Common California natural firewoods and manufactured artificial logs were tested under operating conditions intended to reflect domestic use patterns in the Bay area, which are primarily episodic burning for aesthetic reasons. Emission factors were determined by fuel type, fuel weight, mass emission rates, and energy output, highlighting differences between fuel and combustion facility type. Average PCDD/F emissions factors ranged from 0.25 to 1.4 ng toxic equivalency (TEQ)/kg of wood burned for natural wood fuels and 2.4 ng TEQ/kg for artificial logs. The natural wood emission factors are slightly lower than those which had been estimated for the U.S. inventory. Background-corrected PCBs emitted from woodstove/oak combustion (8370 ng/kg) are 3 orders of magnitude higher in mass than total PCDDs/Fs; however, their toxicity (0.014 ng TEQ/kg) is significantly lower. HxCBz emission factors varied from 13 to 990 ng/kg and were likely fuel- and appliance-specific. Relative PAH concentrations of particle-phase compounds and emission factors were consistent with others' findings. A total of 32 PAH compounds, ranging in concentration from 0.06 to 7 mg/kg, amounted to between 0.12 and 0.38% of the PM mass, depending on the wood and facility type. Preliminary analyses suggest relationships between wood combustion markers and PCDD/F levels. PMID- 12775047 TI - Geochemical signature of contaminated sediment remobilization revealed by spatially resolved X-ray microanalysis of annual rings of Salix nigra. AB - An X-ray microprobe was used to determine the concentration and distribution of Ni, U, and other metals within annual rings of willows (Salix nigra L.) from a former de facto radiological settling basin (Steed Pond; SP) and a depositional environment downstream (Tims Branch; TB) on the Savannah River Site (SRS). Geochemical and historical information about both areas are well documented. Following spillway breaches at SP in 1984 and the early 1990s, TB is inundated with contaminated sediments during storms. Bulk elemental composition of tree cores was determined using ICP-OES. Synchrotron X-rayfluorescence (SXRF) analysis showed that the metal contents of SP and TB cores were an order of magnitude higher than those from a reference site. TB cores were enriched with Ni in 1984 and 1991, corresponding with SP spillway breaches (containing 790 mg kg(-1) Ni in 1991). Cores from SP exhibited an extremely high Ni peak in 1996, approximately 5000 mg kg(-1), even though contaminant levels at SP did not change. The geochemical signature of contaminants recorded in TB annual rings reflected the significant sediment remobilization events consistent with the detailed history of the site, and at concentrations relative to their proximity to the source term. However, physiological processes occurring within impacted trees strongly influence the chronological accuracy of dendroanalysis and must be investigated further. PMID- 12775048 TI - Examination of hydrophobic contaminant adsorption in mineral micropores with grand canonical Monte Carlo simulations. AB - A molecular level understanding of the interactions between hydrophobic organic contaminants (HOCs) and sediments is needed in order to assess contaminant fate in the environment. Grand canonical Monte Carlo simulations were performed to investigate water and trichloroethylene (TCE) adsorption in slit micropores confined by charged and uncharged silica surfaces. Gas-phase single-sorbate simulations with water or TCE were performed as well as mixture simulations of bulk water containing TCE at 1% of its saturation concentration. Gas-phase isosteric heats for water adsorption in the uncharged pores ranged from -40 to 52 kJ/mol, and the densities of the adsorbed water phases were always less than that for bulk water. Gas-phase isosteric heats for water adsorption in the charged pores ranged from -79 to -170 kJ/mol, and the densities of the adsorbed water phases were close to that for bulk water. The isosteric heats and water densities indicated that the uncharged pores were mildly hydrophobic, and the charged pores were very hydrophilic. In mixture simulations of adsorption from solution, the presence of water promoted TCE adsorption in uncharged pores with widths between 14 and 20 A. The isosteric heats for TCE adsorption from solution ranged from -14 to -27 kJ/mol in the uncharged pores and from -9.3 to -50 kJ/mol in the charged pores. Strong attractions to the pore surfaces were significantly diminished after adsorption of the first two monolayers of either adsorbate. Aqueous-phase TCE at a concentration equal to 1% of its saturation concentration was able to completely displace adsorbed water in uncharged pores. Even in highly hydrophilic pores, TCE at this concentration was able to displace up to 50% of the adsorbed water. Apparent differential enthalpies of adsorption determined from the temperature dependence of TCE adsorption isotherms underestimated the magnitude of the true isosteric heats of adsorption by up to 30 kJ/mol. This shows that HOC adsorption enthalpies determined from the temperature dependence of their adsorption isotherms underestimate the true strength of HOC-adsorbent interactions. PMID- 12775049 TI - 3-D structural modeling of humic acids through experimental characterization, computer assisted structure elucidation and atomistic simulations. 1. Chelsea soil humic acid. AB - This paper describes an integrated experimental and computational framework for developing 3-D structural models for humic acids (HAs). This approach combines experimental characterization, computer assisted structure elucidation (CASE), and atomistic simulations to generate all 3-D structural models or a representative sample of these models consistent with the analytical data and bulk thermodynamic/structural properties of HAs. To illustrate this methodology, structural data derived from elemental analysis, diffuse reflectance FT-IR spectroscopy, 1-D/2-D 1H and 13C solution NMR spectroscopy, and electrospray ionization quadrupole time-of-flight mass spectrometry (ESI QqTOF MS) are employed as input to the CASE program SIGNATURE to generate all 3-D structural models for Chelsea soil humic acid (HA). These models are subsequently used as starting 3-D structures to carry out constant temperature-constant pressure molecular dynamics simulations to estimate their bulk densities and Hildebrand solubility parameters. Surprisingly, only a few model isomers are found to exhibit molecular compositions and bulk thermodynamic properties consistent with the experimental data. The simulated 13C NMR spectrum of an equimolar mixture of these model isomers compares favorably with the measured spectrum of Chelsea soil HA. PMID- 12775050 TI - Size-dependent volatility of diesel nanoparticles: chassis dynamometer experiments. AB - We analyzed the size-dependent volatility of nanoparticles in a diameter range of 30-70 nm in diesel exhaust emissions. The test system included a medium-duty diesel truck on a chassis dynamometer, a single-stage dilution tunnel, a tandem differential mobility analyzer (TDMA) equipped with an electric furnace, and a condensation particle counter. The size shifts of monodispersed diesel nanoparticles under changing furnace temperatures were measured by TDMA in the gas phase. Together with the reduction of average particle size and volume, we observed the development of bimodal size distributions resulting from the separation between semivolatile and nonvolatile species as the furnace temperature was increased. While 91-98% of the particles were found to be semivolatile species by total volume during the idling engine condition, only 6 9% were semivolatile during the one-half engine load condition. We also found that smaller particles contained a larger fraction of semivolatile species. PMID- 12775051 TI - Mechanisms affecting the infiltration and distribution of ethanol-blended gasoline in the vadose zone. AB - One- and two-dimensional experiments were conducted to examine differences in the behavior of gasoline and gasohol (10% ethanol by volume) as they infiltrate through the unsaturated zone and spread at the capillary fringe. Ethanol in the spilled gasohol quickly partitions into the residual water in the vadose zone and is retained there as the gasoline continues to infiltrate. Under the conditions tested, over 99% of the ethanol was initially retained in the vadose zone. Depending on the volume of gasoline spilled and the depth to the water table, this causes an increase in the aqueous-phase saturation and relative permeability, thus allowing the ethanol-laden water to drain into the gasoline pool. Under the conditions tested, the presence of ethanol does not have a significant impact on the overall size or shape of the resulting gasoline pool at the capillary fringe. Residual gasoline saturations in the vadose zone were significantly reduced however because of reduced surface and interfacial tensions associated with high ethanol concentrations. The flux of ethanol in the effluent of the column ranged from 1.4 x 10(-4) to 4.5 x 10(-7) g/(cm2 min) with the LNAPL and from 6 x 10(-3) to 3.0 x 10(-4) g/(cm2 min) after water was introduced to simulate rain infiltration. The experimental results presented here illustrate that the dynamic effects of ethanol partitioning into the aqueous phase in the vadose zone create an initial condition that is significantly different than previously understood. PMID- 12775052 TI - Ozone-initiated reactions with mixtures of volatile organic compounds under simulated indoor conditions. AB - This study examines the primary and secondary products resulting from reactions initiated by adding ozone to complex mixtures of volatile organic compounds (VOC). The mixtures were representative of organic species typically found indoors, but the concentrations tended to be higher than normal indoor levels. Each 4-h experiment was conducted in a controlled environmental facility (CEF, 25 m3) ventilated at approximately 1.8 h(-1). The mixture investigated included 23 VOC (no O3), O3/23 VOC, O3/21 VOC (no d-limonene or alpha-pinene), and O3/terpene only (d-limonene and alpha-pinene). The net O3 concentration was approximately 40 ppb in each experiment, and the total organic concentration was 26 mg/m3 for the 23 VOC mixture, 25 mg/m3 for the 21 VOC mixture, and 1.7 mg/m3 for the d-limonene and alpha-pinene mixture. When the 23 VOC were added to the CEF containing no O3, no compounds other than those deliberately introduced were observed. When O3 was added to the CEF containing the 23 VOC mixture, both gas and condensed phase products were found, including aldehydes, organic acids, and submicron particles (140 microg/m3). When O3 was added to the CEF containing the 21 VOC without the two terpenes (O3/21 VOC condition), most of the products that were observed in the O3/23 VOC experiments were no longer present or present at much lower concentrations. Furthermore, the particle mass concentration was 2-7 microg/m3, indistinguishable from the background particle concentration level. When O3 was added to the CEF containing only two terpenes, the results were similar to those in the O3/23 VOC experiments, but the particle mass concentration (190 microg/m3) was higher. The results indicate that (i) O3 reacts with unsaturated alkenes under indoor conditions to generate submicron particles and other potentially irritating species, such as aldehydes and organic acids; (ii) the major chemical transformations that occurred under our experimental conditions were driven by O3/d-limonene and O3/alpha-pinene reactions; and (iii) the hydroxyl radicals (OH) that were generated from the O3/terpene reactions played an important role in the chemical transformations and were responsible for approximately 56-70% of the formaldehyde, almost all of the p-tolualdehyde, and 19-29% of the particle mass generated in these experiments. PMID- 12775053 TI - Effects of FeS on the transformation kinetics of gamma-hexachlorcyclohexane. AB - Distinctly different rates and pathways were observed for abiotic transformation of gamma-hexachlorocyclohexane (gamma-HCH) between homogeneous systems and systems containing FeS solid. The observed half-lives of gamma-HCH decrease from about 1136 and 126 d in homogeneous systems to about 55 and 50 d in heterogeneous systems at pH approximately 6.9 and pH approximately 8.3, respectively. While no appreciable difference in the measured rate was found between the two tested heterogeneous systems of pH 6.9 and pH 8.3, the observed hydrolysis rate constant for the homogeneous systems is highly pH dependent, being constant at approximately 0.0005 d(-1) in pH 2.26-6.73, and increased by an order of magnitude per pH unit from pH 6.73 to pH 12.02. A dehydrohalogenation pathway from gamma-HCH to pentachlorocyclohexene followed by parallel reactions to three trichlorobenzenes is proposed for the hydrolysis reaction in the homogeneous systems. For the heterogeneous systems, the reactions likely follow both the hydrolysis pathway and pathways from gamma-HCH to tetrachlorocyclohexene (TeCCH) and to dichlorocyclohexadiene (DCCD) via dichloroelimination followed by dehydrohalogenation from TeCCH to dichlorobenzenes or from DCCD to chlorobenzene. This study suggests that environmental modeling using hydrolysis rate data measured in homogeneous systems may bear significant inaccuracy for the predicted fate and transport of HCH isomers in highly heterogeneous ecosystems at the regional or mega scales. PMID- 12775054 TI - Transformation of the plant growth regulator daminozide (Alar) and structurally related compounds with CuII ions: oxidation versus hydrolysis. AB - As part of a study of metal ion effects on chemical transformations of nitrogen containing agrochemicals, conversion of daminozide to succinate via cleavage of the hydrazide C-N bond was examined in the presence and absence of divalent metal ions. No conversion was observed in metal ion-free solutions or in the presence of 1.0 mM NiII, ZnII, and PbII. CuII, in contrast, markedly increased rates of daminozide to succinate conversion. Halide ions (CI-, Br-) had no effect on daminozide conversion in the absence of metal ions but markedly increased conversion rates observed in the presence of CuII. The nitrogen-donor ligands ethylenediamine, N-(2-hydroxyethyl)ethylenediamine, and 1,4,7,10 tetraazacyclododecane decreased rates of CuII-facilitated conversion, while 1,5,9 triazacyclododecane actually increased rates of conversion. H NMR and UV spectroscopy provide evidence for the formation of 1:1 CuII-daminozide complexes. Halide ion effects and nitrogen-donor ligand effects point to an oxidative mechanism for CuII-facilitated daminozide breakdown, rather than hydrolysis. The structurally related compound butyric acid 2,2-dimethylhydrazide (BH) is subject to the same CuII-facilitated breakdown via an oxidative mechanism. N,N Dimethylsuccinamic acid (SA), in contrast, breaks down via a hydrolytic mechanism. PMID- 12775055 TI - Alkaline hydrolysis of the cyclic nitramine explosives RDX, HMX, and CL-20: new insights into degradation pathways obtained by the observation of novel intermediates. AB - Hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX, I) and octahydro-1,3,5,7-tetranitro 1,3,5,7-tetrazocine (HMX) hydrolyze at pH > 10 to form end products including NO2 , HCHO, HCOOH, NH3, and N2O, but little information is available on intermediates, apart from the tentatively identified pentahydro-3,5-dinitro-1,3,5 triazacyclohex-1-ene (II). Despite suggestions that RDX and HMX contaminated groundwater could be economically treated via alkaline hydrolysis, the optimization of such a process requires more detailed knowledge of intermediates and degradation pathways. In this study, we hydrolyzed the monocyclic nitramines RDX, MNX (hexahydro-1-nitroso-3,5-dinitro-1,3,5-triazine), and HMX in aqueous solution (pH 10-12.3) and found that nitramine removal was accompanied by formation of 1 molar equiv of nitrite and the accumulation of the key ring cleavage product 4-nitro-2,4-diazabutanal (4-NDAB, O2NNHCH2NHCHO). Most of the remaining C and N content of RDX, MNX, and HMX was found in HCHO, N2O, HCOOH, and NH3. Consequently, we selected RDX as a model compound and hydrolyzed it in aqueous acetonitrile solutions (pH 12.3) in the presence and absence of hydroxypropyl-beta-cyclodextrin (HP-beta-CD) to explore other early intermediates in more detail. We observed a transient LC-MS peak with a [M-H] at 192 Da that was tentatively identified as 4,6-dinitro-2,4,6-triaza-hexanal (O2NNHCH2NNO2CH2NHCHO, III) considered as the hydrolyzed product of II. In addition, we detected another novel intermediate with a [M-H] at 148 Da that was tentatively identified as a hydrolyzed product of III, namely, 5-hydroxy-4-nitro 2,4-diaza-pentanal (HOCH2NNO2CH2NHCHO, IV). Both III and IV can act as precursors to 4-NDAB. In the case of the polycyclic nitramine 2,4,6,8,10,12-hexanitro 2,4,6,8,10,12-hexaazaisowurtzitane (CL-20), denitration (two NO2-) also led to the formation of HCOOH, NH3, and N2O, but neither HCHO nor 4-NDAB were detected. The results provide strong evidence that initial denitration of cyclic nitramines in water is sufficient to cause ring cleavage followed by spontaneous decomposition to form the final products. PMID- 12775056 TI - Formation and extraction of persistent fumigant residues in soils. AB - Fumigants are commonly thought to be short-lived in soil, but residues have been found in soils years following application. In this study, formation and extraction of persistent soil fumigant residues were investigated. Fumigants 1,3 dichloropropene (1,3-D), chloropicrin (CP), and methyl isothiocyanate (MITC) were spiked into Arlington, Glenelg, and Hagerstown soils and incubated for 30 d under controlled conditions. The incubated soils were evaporated for 20 h prior to extraction with a variety of organic solvents at different temperatures. Extraction with acetonitrile in sealed vials at 80 degrees C for 24 h was the most efficient method to recover persistent soil fumigant residues. At application rates of 1000-1700 mg (kg of soil)(-1), persistent residues of 1,3-D, CP, and MITC in the three soils ranged from 5 to 67 mg kg(-1). The residue content increased with application rate, correlated positively with soil silt content, decreased dramatically as indigenous organic matter (OM) was removed, and changed little with external OM addition. Adsorption to clay surfaces was not important in fumigant retention, while pulverization of soil aggregates significantly decreased persistent fumigant residues. The results suggest that persistent fumigant residues are retained in soil intra-aggregate micropores resulting from binding clay flocs and silt particles by humic substances. PMID- 12775057 TI - Inhibition of bacterial U(VI) reduction by calcium. AB - The rapid kinetics of bacterial U(VI) reduction and low solubility of uraninite (UO2,cr) make this process an attractive option for removing uranium from groundwater. Nevertheless, conditions that may promote or inhibit U(VI) reduction are not well-defined. Recent descriptions of Ca-UO2-CO3 complexes indicate that these species may dominate the aqueous speciation of U(VI) in many environments. We monitored the bacterial reduction of U(VI) in bicarbonate-buffered solution in the presence and absence of Ca. XAFS measurements confirmed the presence of a Ca U(VI)-C03 complex in the initial solutions containing calcium. Calcium, at millimolar concentrations (0.45-5 mM), caused a significant decrease in the rate and extent of bacterial U(VI) reduction. Both facultative (Shewanella putrefaciens strain CN32) and obligate (Desulfovibrio desulfuricans, Geobacter sulfurreducens) anaerobic bacteria were affected by the presence of calcium. Reduction of U(VI) ceased when the calculated system Eh reached -0.046 +/- 0.001 V, based on the Ca2UO2(CO3)3 --> UO2,cr couple. The results are consistent with the hypothesis that U is a less energetically favorable electron acceptor when the Ca-UO2-CO3 complexes are present. The results do not support Ca inhibition caused by direct interactions with the cells or with the electron donor as the reduction of fumarate or Tc(VII)O4- under identical conditions was unaffected by the presence of Ca. PMID- 12775058 TI - Uptake and reduction of Cr(VI) to Cr(III) by mesquite (Prosopis spp.): chromate plant interaction in hydroponics and solid media studied using XAS. AB - Chromium (Cr) is a well-established carcinogen that is a contaminant at half of the EPA Superfund sites in the United States. Two separate studies were performed to investigate the possibility that mesquite (Prosopis spp.), which is an indigenous desert plant species, can remove Cr from the environment via active transport systems to the aerial portions of the plant. The first study was performed by growing mesquite on solid media (agar) at Cr(VI) concentrations of 75 and 125 ppm. The accumulation found in the leaves under the present conditions indicated that mesquite could be classified as a hyperaccumulator of chromium. The second study was conducted to investigate the differences between the type of Cr ligand involved in Cr uptake with agar and hydroponic cultures. We used X-ray absorption spectroscopy (XAS) to determine the mechanisms involved in the uptake and binding of Cr(VI) in live mesquite tissue. The XAS results for this study showed that some of the supplied Cr(VI) was uptaken by the mesquite roots; however, the data analyses of the plant tissues demonstrated that it was fully reduced to Cr(III) in the leaf tissues. Experiments are currently being performed to evaluate the behavior of the Mesquite plant using lower Cr concentrations. PMID- 12775059 TI - Formation of carbonyl sulfide by the reaction of carbon monoxide and inorganic polysulfides. AB - OCS formation by the reaction of inorganic polysulfides with carbon monoxide, which are both abundant in natural aquatic systems, was studied as a model abiotic route for OCS formation in the dark. The net OCS accumulation rate was a function of a bimolecular formation reaction and simultaneous OCS hydrolysis kinetics. The reaction of polysulfides with CO in the dark was found to be first order with respect to CO concentration and first order with respect to the molar sum of the polysulfide species generated by the disproportionation of the dissolved polysulfide precursors. The pH dependence of the OCS production rate was controlled by the pH-dependent disproportionation of polysulfide precursors. Lower temperatures, intermediate redox potentials, and moderately basic pH conditions increase the steady-state concentration of OCS. The speciation of polysulfides in aqueous solutions is still disputed. Some authors claim that hexasulfide is one of the dominant species while others believe that pentasulfide is the largest sulfide species in aqueous systems. Despite the disagreement between different models for speciation of polysulfides, the proposed rate law agreed very well with the thermodynamic data based on four and on five polysulfide species, with only minor differences in the preexponential kinetic coefficients. PMID- 12775060 TI - Source apportionment of polycyclic aromatic hydrocarbons in the urban atmosphere: a comparison of three methods. AB - Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous pollutants in urban atmospheres. Several PAHs are known carcinogens or are the precursors to carcinogenic daughter compounds. Understanding the contributions of the various emission sources is critical to appropriately managing PAH levels in the environment. The sources of PAHs to ambient air in Baltimore, MD, were determined by using three source apportionment methods, principal component analysis with multiple linear regression, UNMIX, and positive matrix factorization. Determining the source apportionment through multiple techniques mitigates weaknesses in individual methods and strengthens the overlapping conclusions. Overall source contributions compare well among methods. Vehicles, both diesel and gasoline, contribute on average 16-26%, coal 28-36%, oil 15-23%, and wood/other having the greatest disparity of 23-35% of the total (gas- plus particle-phase) PAHs. Seasonal trends were found for both coal and oil. Coal was the dominate PAH source during the summer while oil dominated during the winter. Positive matrix factorization was the only method to segregate diesel from gasoline sources. These methods indicate the number and relative strength of PAH sources to the ambient urban atmosphere. As with all source apportionment techniques, these methods require the user to objectively interpret the resulting source profiles. PMID- 12775061 TI - Risk-based approach to evaluate the public health benefit of additional wastewater treatment. AB - The City of Stockton, CA operates a wastewater treatment facility that discharges tertiary treated effluent during the summer and secondary treated effluent during the winter to the San Joaquin River. Investigations were carried out between 1996 and 2002 to provide insight regarding the potential public health benefit that may be provided by year-round tertiary treatment. A hydraulic model of the San Joaquin River and a dynamic disease transmission model integrated a wide array of disparate data to estimate the level of viral gastroenteritis in the population under the two treatment scenarios. The results of the investigation suggest that risk of viral gastroenteritis attributable to the treatment facility under the existing treatment scheme is several orders of magnitude below the 8-14 illnesses per 1000 recreation events considered tolerable by U.S. EPA, and winter tertiary treatment would further reduce the existing risk by approximately 15-50%. The methodologies employed herein are applicable to other watersheds where additional water treatment is being considered to address public health concerns from recreation in receiving waters. PMID- 12775062 TI - Assessment of a sequential extraction procedure for perturbed lead-contaminated samples with and without phosphorus amendments. AB - Sequential extraction procedures are used to determine the solid-phase association in which elements of interest exist in soil and sediment matrixes. Foundational work by Tessier et al. (Tessier, A.; Campbell, P. G. C.; Bisson, M. Anal. Chem. 1979, 51, 844-851) has found widespread acceptance and has been employed as an operational definition for metal speciation in solid matrixes. However, a major obstacle confronting sequential extraction procedures is species alteration of extracted metals before, during, and after separation of solids from solution. If this occurs, the results obtained from sequential extraction do not provide an accurate account of metal speciation within the matrix because the metal forms are altered from their field state. Many researchers dismiss this drawback since several sorption and precipitation processes are believed to occur at time scales much longer than any particular extraction step. This assumption may not be valid. The objectives of this study were to investigate the potential formation of pyromorphite (Pb5(PO4)3Cl) during the sequential extraction steps of Pb-spiked samples with and without calcium phosphate amendments and to examine the differences in the operationally defined distribution of Pb in samples with and without the presence of P. The systems that were examined in the absence of phosphate behaved, for the most part, adequately according to the operational definitions of the extraction procedure. However, when the samples were amended with phosphate, results were drastically changed with a significant shift of extractable Pb to the residual phase. This redistribution was due to pyromorphite formation during the extraction procedure as confirmed by X-ray diffraction and X ray absorption (XAS) spectroscopies. These results indicate that sequential extraction methods may not be suitable for Pb speciation in perturbed environmental systems (i.e., fertilized agricultural soils or amended contaminated soils) and that rigorous interpretation should be avoided, if not supported by methods to definitively prove metal speciation (e.g., XAS). PMID- 12775063 TI - Development of an ultrarapid one-step fluorescence immunochromatographic assay system for the quantification of microcystins. AB - Microcystins are a family of potent toxic oligopeptides produced by freshwater cyanobacteria genera and have been a great threat to the welfare of humans and animals. There has been a great demand for developing a fast and convenient analytical method to detect microcystins. Recently, direct competition ELISA using monoclonal or polyclonal antibody has become the prevailing method for detecting microcystins. In this study, we report rapid quantification methods of microcystins using fluorescence for a detection signal and a lateral-flow-type immunochromatography as a separation system. The assay systems consist of a test strip housed in a disposable cartridge and a portable laser-fluorescence scanner. The components of a test strip are as follows: a nitrocellulose membrane, a sample pad, an absorption pad, and a backing card. A fluorescence scanner was designed to fit the cartridge and to quantify the distribution of the fluorescence intensity along the strip. When the calibration curve for an antibody-immobilized system was determined, a good linearity was displayed in the range from 125 to 2000 pg/mL of microcystin-LR. The linear-regression coefficient (R) was 0.938 between relative fluorescence intensity and the microcystin concentration. The limit of detection was determined to be 95.38 pg/mL. We then designed another biosensor system by changing an experimental format from the competition type to the inhibition type. When compared to the antibody immobilized system, the antigen-immobilized assay detected a lower level of microcystin but did not discern microcystin-LR above 1000 pg/mL. The detection of limit for the antigen-immobilized system was 47.23 pg/mL. The linear regression coefficient (R) in the antigen-immobilized system equaled to 0.927. The reproducibility in the antigen-immobilized system was good through the entire range. The reproducibility in the antibody-immobilized system was relatively poor when compared to a MC-immobilized system. However, it still registered in the acceptable range of 7.32-9.91% except for the extreme ends of the MC concentration. Finally, surface water was tested to check for potential matrix interference. The calibration curve displayed a similar pattern as did those for other matrixes, including PBS and tap water, although its sensitivity was a little less due to the interference with certain components in the surface water. Overall, either of the biosensor systems can be used as a useful on-site detection tool for checking drinking water or surface water for microcystins. The laser-fluorescence scanner we developed is relatively small, transportable, and easy to use. Thus, the samples can be analyzed for microcystins at the test site using a real-time base within 15 min without having to bring the samples back to the laboratory. PMID- 12775064 TI - Dissolved oxygen imaging in a porous medium to investigate biodegradation in a plume with limited electron acceptor supply. AB - A novel combination of noninvasive imaging with an oxygen sensitive fluorescent indicator was developed to investigate the biodegradation processes occurring at the fringe of a solute plume, where the supply of oxygen was limited. A thin transparent porous matrix (156 x 120 x 3 mm) was made from quartz plates and quartz sand (212-300 microm) and enriched with acetate-degrading bacteria. A degrading plume developed from a continuous acetate source in the uniform flow field containing dissolved oxygen. Ruthenium (II)-dichlorotris(1,10 phenanthroline) (Ru(phen)3Cl2), a water-soluble fluorescent dye, was used as an indicator of dissolved oxygen. The fluorescence intensity was dependent on the concentration of oxygen because the dissolved oxygen acted as collisional quencher. The oxygen distribution was interpreted from images recorded by a CCD camera. These two-dimensional experimental results showed quantitatively how the oxygen concentrations decreased strongly at the narrow plume fringe and that oxygen was depleted at the core of the plume. Separately, dispersivity was measured in a series of nonreactive transport experiments, and biodegradation parameters were evaluated by batch experiments. Two-dimensional numerical simulations with MT3D/RT3D used these parameters, and the predicted oxygen distributions were compared with the experimental results. This measurement method provides a novel approach to investigate details of solute transport and biodegradation in porous media. PMID- 12775065 TI - Selective remediation of contaminated sites using a two-level multiphase strategy and geostatistics. AB - Selective soil remediation aims to reduce costs by cleaning only the fraction of an exposure unit (EU) necessary to lower the average concentration below the regulatory threshold. This approach requires a prior stratification of each EU into smaller remediation units (RU) which are then selected according to various criteria. This paper presents a geostatistical framework to account for uncertainties attached to both RU and EU average concentrations in selective remediation. The selection of RUs is based on their impact on the postremediation probability for the EU average concentration to exceed the regulatory threshold, which is assessed using geostatistical stochastic simulation. Application of the technique to a set of 600 dioxin concentrations collected at Piazza Road EPA Superfund site in Missouri shows a substantial decrease in the number of RU remediated compared with single phase remediation. The lower remediation costs achieved by the new strategy are obtained to the detriment of a higher risk of false negatives, yet for this data set this risk remains below the 5% rate set by EPA region 7. PMID- 12775066 TI - Changes in interfacial tension of chlorinated solvents following flow through U.K. soils and shallow aquifer material. AB - The interfacial tension (IFT) that arises at the interface between water and an immiscible organic liquid is a key parameter affecting the transport and subsequent fate of the organic liquid in water-saturated porous media. In this paper, data are presented that show how contact between a range of soil types and chlorinated hydrocarbon solvent (CHS) dense nonaqueous phase liquids (DNAPLs) can affect DNAPL/water IFT values. The soils examined are indicative of U.K. soil types and shallow aquifer materials. The solvents investigated were tetrachloroethylene (PCE) and trichloroethylene (TCE). Lab grade, recovered field DNAPL and industrial waste chlorinated solvent mixtures were used. The data from batch and column experiments invariably revealed that water/DNAPL IFT values change following contact with unsaturated soils. In the majority of cases, the IFT values increase following soil exposure. However, after contact with an organic-rich soil, the IFT of the lab grade solvents decreased. The experimental evidence suggests that these reductions are linked to the removal of organic material from the soil and its subsequent incorporation into the solvent IFT increases in the case of lab solvents are shown to be linked to the removal of stabilizers (added by the manufacturers to obviate degradation) that are removed by adsorption to soil mineral surfaces. Similarly, it is conjectured that adsorption of surface-active compounds from the industrial waste samples to soil surfaces is responsible for increases in the IFT in these samples. Finally, it was observed that invading CHSs are capable of dissolving and subsequently mobilizing in-situ soil contaminants. GC/MS analysis revealed these mobilized soil contaminants to be polyaromatic hydrocarbons and phthalate esters. PMID- 12775067 TI - Synergy of combining sonolysis and photocatalysis in the degradation and mineralization of chlorinated aromatic compounds. AB - Merits of using advanced oxidation processes such as sonolysis and photocatalysis as well as a combination of the two have been explored using model herbicides such as 2,4-dichlorophenoxy acetic acid and 2,4-dichlorophenoxypropionic acid and the chlorinated phenols 2,4-dichlorophenol and 2,4,6-trichlorophenol. Whereas sonolysis is quite effective in the initial degradation of chlorinated aromatic molecules, complete mineralization is difficult to achieve. Photocatalysis is selective toward the degradation of polar compounds but causes the build up of undesirable chemical intermediates. In contrast to sonolytic degradation, photocatalysis is very effective toward achieving complete mineralization. By simultaneously carrying out high-frequency sonolysis and photocatalysis we have succeeded in achieving faster and complete mineralization with no build up of toxic intermediates even at very low catalyst loadings. The synergy of combining the two advanced oxidation processes is discussed. PMID- 12775068 TI - Experimental and model comparisons of low- and medium-pressure Hg lamps for the direct and H2O2 assisted UV photodegradation of N-nitrosodimethylamine in simulated drinking water. AB - Both low- and medium-pressure Hg lamps (LP and MP, respectively) were used as ultraviolet light (UV) sources to destroy N-nitrosodimethylamine in a synthetic "natural" water. The lamp performances were directly compared via the UV fluence based rate constants, which demonstrates that LP and MP have virtually identical photonic efficiencies (fluence-based rate constants of 2.29E-3 and 2.35E-3 cm2/mJ, respectively). This indicates that the quantum yield for NDMA photolysis is independent of wavelength in the UVC region: a value of 0.30 mol/einstein is found at pH 8.1. Addition of 100 mg/L of H2O2 leads to a 30% increase in the LP fluence-based rate constant but does not alter the MP rate constant, likely due to the tradeoff between light screening by H2O2 and additional radical based degradation. However, in terms of the time-based rate constant, this level of H2O2 slightly enhances the LP performance but hinders the MP performance, suggesting that H2O2 is of little or no economic benefit for NDMA removal by UV. All these effects are explained by modeling the photochemistry according to standard equations. The model predicts that H2O2 may enhance NDMA removal for short optical path lengths but that light-screening by H2O2 may decrease the removal rates for optical path lengths typical of those found in UV reactors. PMID- 12775069 TI - Photolysis of ozone in aqueous solutions in the presence of tertiary butanol. AB - The ozone decomposition quantum yield (phi) in millimolar and higher concentration aqueous tertiary butanol solution is 0.64 +/- 0.05 (observed over a wavelength range from 250 to 280 nm) and rises toward lower tertiary butanol concentrations (phi approximately 1.5 at 10(-5) M at pH 2) on account of the onset of the well-known *OH-radical-induced chain reaction. The destruction of the organic is initiated by hydrogen-atom abstraction through OH radicals which are produced via the reaction of the photolytically generated O(1D) with the solvent water at a quantum yield of phi(*OH) of about 0.1. There is no decomposition of ozone in the dark on the time scale of the photolysis experiment. The efficiency of tertiary butanol destruction with respect to ozone consumption ([O3]0 = 3 x 10(-4) M), defined by the ratio delta[t-BuOH]/delta[O3], termed eta(t-BuOH), is 0.26 at millimolar tertiary butanol concentrations, determined at the stage of essentially complete ozone consumption. It diminishes toward lower tertiary butanol concentrations (delta[t-BuOH]/delta[O3] approximately 0.17 at [t-BuOH]0 = 1 x 10(-4) M). Part of the effect of the ozone, apart from being a source of *OH radicals, rests on the intervention of HO2*/O2*- which is produced in the course of the peroxyl-radical chemistry of the tertiary butanol in this dioxygen-saturated environment and converted into further *OH radical by reaction with ozone. Moreover in this system, organic free radicals and peroxyl radicals react with the ozone. On the basis of the experimental and mechanistic-simulation data, the quantum yield of direct (by hv) ozone cleavage in aqueous solution is estimated at about 0.5. PMID- 12775070 TI - Kinetics of diesel nanoparticle oxidation. AB - The technique of high-temperature oxidation tandem differential mobility analysis has been applied to the study of diesel nanoparticle oxidation. The oxidation rates in air of diesel nanoparticles sampled directly from the exhaust stream of a medium-duty diesel engine were measured over the temperature range of 800-1140 degrees C using online aerosol techniques. Three particle sizes (40, 90, and 130 nm mobility diameter) generated under engine load conditions of 10, 50, and 75% were investigated. The results show significant differences in the behavior of the 10% load particles as compared to the 50 and 75% load particles. The 10% load particles show greater size decrease at temperatures below 500 degrees C and significant size decrease at temperatures between 500 and 1000 degrees C in a non oxidative environment, indicating release of adsorbed volatile material or thermally induced rearrangement of the agglomerate structure. Activation energies determined are 114, 109, and 108 kJ mol(-1) for the 10, 50, and 75% load particles, respectively. These activation energies are lower than for flame soot (Higgins et al. J. Phys. Chem. A 2002, 106, 96), but the preexponential factors are lower by 3 orders of magnitude, and the overall oxidation rates are slower by up to a factor of 4 over the temperature range studied. Possible reasons for the differences are discussed in the text. PMID- 12775071 TI - Supercritical water oxidation of sulfide. AB - Supercritical water oxidation (SCWO) of sulfide wastewater with oxygen as the oxidant was investigated using ammonium sulfide solution as the model waste. The experiments, which were conducted in a bench-scale continuous SCWO installation, indicated that S2- could be completely oxidized to SO4(2-) within the residence time of less than 10 s under the supercritical water conditions at 698.2-773.2 K and 22.0 MPa-30.0 MPa. The reaction pathway of S2- in SCWO was determined to be S2- --> S2O3(2-) --> SO3(2-) --> SO4(2-). The effects of temperature, pressure, and oxygen content on SCWO of S2- were also studied. The kinetic model based on this pathway was established with the reaction rate constants and orders found by fitting the experimental data. The dependence of the rate constants on the temperature can be described with Arrhenius' law. The rate constants are dependent on the density of SCW, and the dependence is logarithmically linear. PMID- 12775072 TI - Key parameters for de novo formation of polychlorinated dibenzo-p-dioxins and dibenzofurans. AB - De novo formation of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDDs and PCDFs) was investigated in an Entrained Flow Reactor (EFR) to simulate combustion conditions. The parameters investigated were carbon content and nature in fly ash; type of gas-phase environment (oxidative versus reducing conditions) influence of combustion gases such as water, carbon monoxide, and carbon dioxide; amount of gas-phase chlorine; reaction temperature (250-600 degrees C); and reaction time (minutes vs hours). The comprehensive data set was further evaluated with principal component analysis (PCA) to statistically determine the role and importance of each parameter for de novo formation of PCDDs and PCDFs. Results revealed that an initial fast de novo formation occurs within the first minutes with a formation rate in the orders of hundreds of pmol per minutes; however, the reactivity of the ash was found to decline with time. An average formation rate as low as 3 pmol/min was measured after 6 h. The slower de novo formation of PCDDs and PCDFs was found to be through different reaction mechanisms and, thus, controlled by different parameters. The amount of Cl2 in the gas phase was observed to be an important parameter for PCDFs formation; meanwhile the levels of O2 were not found to be a PCDF rate controlling parameter. The formation rate of PCDDs was significantly lower than the PCDFs, and two mechanisms appear to be controlling the formation, one depending on the amount of O2 and one on the amount of Cl2 present in the gas phase. Overall the most significant parameter for the rate of formation for both PCDDs and PCDFs was revealed to be the reaction temperature. A maximum rate of formation was observed between 300-400 degrees C for the PCDDs and 400-500 degrees C for the PCDFs. PMID- 12775073 TI - Passive in situ remediation of metal-polluted water with caustic magnesia: evidence from column experiments. AB - Passive remediation consists of a permeable system that enables the water to pass through while retaining metals by means of biogeochemical reactions. Conventional passive treatments are based on calcite dissolution. This increases the pH to values between 6 and 7, which are insufficiently high to precipitate divalent metals. Alternative treatments are based on sulfate reduction with organic matter in order to precipitate metal sulfides. However, redox reactions are usually too slow to treat large groundwater flows as currently found in gravel aquifers (>50 m/a). Caustic magnesia obtained from calcination of magnesium carbonate was tested as an alternative material to devising passive remediation systems. Caustic magnesia reacts with water to form magnesium hydroxide, which dissolves, increasing the pH to values higher than 8.5. Then zinc and lead are mainly precipitated as hydroxides, copper is precipitated as hydroxysulfate, and manganese(II) is oxidized and precipitated as manganese(III) oxides. Thus, metal concentrations as high as 75 mg/L in the inflowing water are depleted to values below 0.04 mg/L. Magnesia dissolution is sufficiently fast to treat flows as high as 100 m/a. The new precipitates may lead to a permeability drop in the porous treating system. Mixtures of caustic magnesia and an inert material such as silica sand (approximately 50% of each) have been shown to be as reactive as pure magnesia and permeable for a longer time (more than 10 months and 1000 pore vol). PMID- 12775074 TI - Sulfur dioxide treatment from flue gases using a biotrickling filter-bioreactor system. AB - Complete treatment of sulfur dioxide (SO2) from flue gases in a two-stage process consisting of a biotrickling filter followed by biological post-treatment unit was investigated. The biotrickling filter could remove 100% of influent SO2 from simulated flue gas at an empty bed residence time of 6 s for a concentration range of 300-1000 ppm(v). All the absorbed SO2 was recovered in the biotrickling filter liquid effluent as sulfite (a product of chemical reaction of SO2) and sulfate (product of biological oxidation of sulfite). The biotrickling filter liquid effluent was further processed biologically in a single post-treatment unit consisting of a combined anaerobic and microaerophilic reactor for the simultaneous reduction of sulfate and sulfite to sulfide and oxidation of sulfide to elemental sulfur. The post-treatment unit could effectively treat the biotrickling filter effluent and produce elemental sulfur. The sulfur production efficiency of the reactor reached about 80% of the SO2 treated. This new biological treatment system seems to be a promising alternative for flue gas desulfurization. PMID- 12775075 TI - Rate-limited solubilization of multicomponent nonaqueous-phase liquids by flushing with cosolvents and surfactants: modeling data from laboratory and field experiments. AB - The impact of rate-limited mass transfer on in-situ cosolvent and single-phase microemulsion flushing for the solubilization of multicomponent nonaqueous-phase liquids (NAPLs) was investigated. Laboratory and field data from both cosolvent and microemulsion flushing studies were modeled using a one-dimensional flow, enhanced-dissolution code. Transport of the flushing agents was described with a one-dimensional advection-dispersion model; more complex heterogeneities encountered at the field scale were characterized using the superposition of two solute transport equations. Cosolvent and microemulsion flushing solubilize NAPLs by different mechanisms. The modeling results presented here show that nonequilibrium effects influence both of these processes differently. Solubilization of individual constituents by cosolvents was limited by the rate of diffusion or transport of the component through the organic phase, while the rate-limiting step for microemulsification was independent of the NAPL constituent and was likely external to the organic phase. These data indicate that by accounting for field-scale hydrodynamic variability, laboratory-measured nonequilibrium parameters may be used to accurately predict field-scale nonequilibrium NAPL solubilization. Finally, the effects of field-scale media heterogeneity are likely to dominate those of weakly rate-limited dissolution, and accurate characterization of the former may be sufficient for adequate prediction of field-scale NAPL solubilization. PMID- 12775076 TI - Comparing Nafion and ceramic separators used in electrochemical purification of spent chromium plating solutions: cationic impurity removal and transport. AB - This study focuses on the electrolytic regeneration of spent chromium plating solutions. These solutions contain a significant amount of chromium and a lesser amount of other heavy metals, which makes them a significant environmental concern and an obvious target for recycling and reuse. The type of separator used is extremely critical to the performance of the process because they are the major resistance in the transport-related impurity (Cu(II), Ni(II), and Fe(III)) removals from contaminated chromic acid solutions. A Nafion 117 membrane and a ceramic diaphragm separator traditionally used in the industry were tested for comparison. It was found that the mobilities of Cu(II) and Ni(II) were similar and higher than that of Fe(III) using both separators. The mobility of each cation was smaller in the Nafion membrane than in the ceramic diaphragm. The measured conductivity of the ceramic diaphragm was slightly higher than that of Nafion membrane. However, the Nafion membrane was much thinner than the ceramic diaphragm resulting in the system using the Nafion membrane having higher impurity removal rates than the system using the ceramic diaphragm. The removal rates were approximately equal for Cu(II) and Ni(II) and lowest for Fe(III). Both current and initial concentration affected the removal rates of the impurities. Modeling results indicated that a system using a Nafion separator and a small catholyte/anolyte volume ratio was better than a system using a ceramic separator for removing impurities from concentrated plating solutions if the impurities transported into the catholyte are deposited or precipitated. PMID- 12775077 TI - Mechanism of high-temperature CO2 sorption on lithium zirconate. AB - Lithium zirconate (Li2ZrO3) is one of the most promising materials for CO2 separation from flue gas at high temperature. This material is known to be able to absorb a large amount of CO2 at around 400-700 degrees C. However, the mechanism of the CO2 sorption/desorption process on Li2ZrO3 is not known yet. In this study, we examined the CO2 sorption/desorption mechanism on Li2ZrO3 by analyzing the phase and microstructure change of Li2ZrO3 during the CO2 sorption/desorption process with the help of thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), and X-ray diffraction (XRD) analyses. Li2ZrO3 powders were prepared from lithium carbonate (Li2CO3) and zirconium oxide (ZrO2) by the solid-state method, and the CO2 sorption/desorption property was examined by TGA. It was shown that pure Li2ZrO3 absorbs a large amount of CO2 at high temperature with a slow sorption rate. Addition of potassium carbonate (K2CO3) and Li2CO3 in the Li2ZrO3 remarkably improves the CO2 sorption rate of the Li2ZrO3 materials. DSC analysis for the CO2 sorption process indicates that doped lithium/potassium carbonate is in the liquid state during the CO2 sorption process and plays an important role in improving the CO2 uptake rate. XRD analysis for phase and structure change during the sorption/desorption process shows that the reaction between Li2ZrO3 and CO2 is reversible. Considering all data obtained in this study, we proposed a double-shell model to describe the mechanism of the CO2 sorption/desorption on both pure and modified Li2ZrO3. PMID- 12775078 TI - Compilation and application of Japanese inventories for energy consumption and air pollutant emissions using input-output tables. AB - Preparing emission inventories is essential to the assessment and management of our environment. In this study, Japanese air pollutant emissions, energy consumption, and CO2 emissions categorized by approximately 400 sectors (as classified by Japanese input-output tables in 1995) were estimated, and the contributions of each sector to the total amounts were analyzed. The air pollutants examined were nitrogen oxides (NOx), sulfur oxides (SOx), and suspended particulate matter (SPM). Consumptions of about 20 fossil fuels and five other fuels were estimated according to sector. Air pollutant emission factors for stationary sources were calculated from the results of a survey on air pollution prevention in Japan. Pollutant emissions from mobile sources were estimated taking into consideration vehicle types, traveling speeds, and distances. This work also counted energy supply and emissions from seven nonfossil fuel sources, including nonthermal electric power, and CO2 emissions from limestone (for example, during cement production). The total energy consumption in 1995 was concluded to be 18.3 EJ, and the annual total emissions of CO2, NOx, SOx, and SPM were, respectively, 343 Mt-C, 3.51 Mt, 1.87 Mt, and 0.32 Mt. An input-output analysis of the emission inventories was used to calculate the amounts of energy consumption and emissions induced in each sector by the economic final demand. PMID- 12775079 TI - Theoretical calculation of product contents: battery and cathode ray tube examples. AB - Most product environmental assessments are based on manufacturer-supplied data on the material content of the product. This paper explores the potential for the material content of key components to be estimated with theoretical calculations. Two examples, the amount of cadmium in a nickel-cadmium battery and the amount of lead in a TV or computer CRT monitor, are developed. Both an upper and a lower limit on the amount of cadmium in a nickel-cadmium battery are calculated on the basis of the battery's chemical reaction. The amount of lead shielding needed in a TV or CRT computer monitor is estimated on the basis of the potential difference through which electrons are accelerated and the absorption length of photons in lead. Such calculations can be used as benchmarks in product environmental assessments, providing validation of manufacturer-supplied data and providing insight into the composition and design of products. PMID- 12775080 TI - Comment on "The West Falmouth oil spill after thirty years: the persistence of petroleum hydrocarbons in marsh sediments". PMID- 12775081 TI - Comment on "Electrostatic sampler for semivolatile aerosols: chemical artifacts" and "Filter and electrostatic samplers for semivolatile aerosols: physical artifacts". PMID- 12775082 TI - Findings from the International Adult Literacy Survey on the incidence and correlates of learning disabilities in New Zealand: is something rotten in the state of New Zealand?. AB - New Zealand data from the International Adult Literacy Survey were analysed to examine the incidence and correlates of self-reported specific reading learning disability (SRLD). The results showed that 7.7% of New Zealand adults reported having had a learning disability. The ratio of males to females with SRLD was 3:2. Between 40% and 50% of New Zealand adults performed below the minimum level of proficiency required for meeting the complex demands of everyday life in knowledge-based societies. For adults with SRLD, around 80% performed below the minimum level, and the literacy proficiency of adults with SRLD in younger age bands appears to have declined since the early 1960s. Almost 100% of adults with SRLD in the 16-20 years age range performed below the minimum level for document and quantitative literacy and 92% for prose literacy. Compared to non-SRLD adults, those with SRLD were found to leave school earlier, engage more often in manual occupations, are more frequently unemployed, and rely on more state assistance to bring their income levels closer to the levels enjoyed by non-SRLD adults. The results are discussed in terms of SRLD not being officially recognised or provided for in New Zealand, the lack of appropriate remedial provisions for children who experience difficulties with reading, and the effects of a strong whole language orientated approach to literacy instruction in schools that has been in place since 1963. PMID- 12775083 TI - The International Adult Literacy Survey in Britain: impact on policy and practice. AB - There is increasing concern for the skills of the workforce in the UK and elsewhere, but despite this concern until recently there has been little information available which objectively measures basic skills in adults. In this paper, evidence derived from the prose scale of the International Adult Literacy Survey (IALS, 1996) is outlined, with emphasis on the performance of adults at the lowest levels, 1 and 2 in the United Kingdom. A new analysis based on the 183 adults who self-reported learning disabilities demonstrates that over 50% of this group perform at level 1 on the prose scale. Over 60% report that these disabilities persist into adult life, although this number falls to 50% in the youngest age group, reflecting changes in recognition of learning disabilities within the education system. The paper concludes with a case study of the redefinition of basic skill levels in Britain based on the IALS levels. The impact of the IALS findings on policy and practice, and in particular through the recommendations of the Moser report, are discussed. PMID- 12775084 TI - Commentaries on the Reynolds et al. article. PMID- 12775085 TI - Evaluation of an exercise based treatment for children with reading difficulties. PMID- 12775086 TI - A critique of claims from Reynolds, Nicolson & Hambly (2003) that DDAT is an effective treatment for children with reading difficulties--'lies, damned lies and (inappropriate) statistics'? AB - Reynolds, Nicolson and Hambly (Dyslexia 2003; 9: 48) report a study to evaluate the effectiveness of dyslexia dyspraxia attention deficit treatment (DDAT)-an exercise-based treatment for children with reading difficulties. They claim that DDAT is an effective treatment with positive effects on the cognitive skills underlying literacy and the reading process. We outline the numerous methodological and statistical problems with this study and conclude that it provides no evidence that DDAT is an effective form of treatment for children with reading difficulties. PMID- 12775087 TI - Determination of cut-off values for leucocytes and bacteria for urine flow cytometer (UF-100) in urinary tract infections. AB - Leucocytes and bacteria in urine are the important indices for diagnosing urinary tract infections. Urine specimens from 260 patients were examined for leucocytes and bacteria with the UF-100 flow cytometer (Sysmex Corporation, Japan). The results obtained with the UF-100 were compared with those obtained by bacteriologic cultures of the same urine samples. Bacteriologic samples were found to be positive in 48 urine samples. The best detected cut-off value for bacteria is 3800/microL (senstivity = 73%, specificity = 78%) and 25/microL for leucocytes (sensitivity = 73%, specificity = 92%). In addition, comparison of leucocyte count by manually and with UF-100 shows good correlation (r = 0.91). A flow cytometric urinalysis analyzer operates in a time saving manner, increases throughput and decrease microscopy review rates. It also provides a reliable information to clinicians prior to bacteriologic culture results, in terms of leucocyte and bacteria count. PMID- 12775088 TI - Hydronephrotic obstructed kidney mimicking a congenital multicystic kidney: case report with review of literature. AB - OBJECTIVE: To report a case of obstructed hydronephrotic kidney mimicking a multicystic kidney and to review the literature regarding differentiation of the hydronephrotic variant of multicystic kidney from the obstructed hydronephronic kidney. To suggest a possible algorithm in distinguishing them. METHODS: We have reported a case of a 35-year-old male who presented with dull aching pain and a palpable right-sided cystic flank mass of several years duration. The initial workup suggested a nonfunctioning multicystic kidney while the operative findings and histopathology were suggestive of an obstructed hydroenphrotic kidney with pyelonephritic changes. We searched the literature using the key words hydronephrotic dysplastic kidney and multicystic kidney. RESULTS: A detailed literature search did not reveal any such publication describing the differentiation of the hydronephrotic multicystic dysplastic kidney from the obstructed hydronephrotic kidney of pelviureteral obstruction. We reviewed the existing literature on this subject, on the basis of which, we have suggested a six-stepladder approach to distinguish such cases. CONCLUSION: By using the 6 step ladder protocol algorithm suggested by us one can attempt to distinguish the hydronephrotic variant of multicystic dysplastic kidney from the hydronephrotic kidney due to pelviureteral obstruction in patients presenting with a symptomatic cystic flank masses of renal origin. Differentiation between the two may be difficult at times due to the medial/central placement of cysts in the former. This is necessary since renal salvage may be possible in the latter while timely nephrectomy may be considered in the former to prevent against the hazards of leaving behind a dysplastic kidney in situ. PMID- 12775089 TI - Simple method of residual stone retrieval through the nephrostomy catheter after PCNL: point of technique. AB - A simple technique of removal of residual stone fragments in patients who have undergone percutaneous nephrolithotripsy (PCNL) and have a nephrostomy tube in situ is described. Under local anesthesia and sedation a rigid 8.5 F ureteroscope is passed through the nephrostomy tube and the stones retrieved. This technique is indicated in patients who have 1-3 residual fragments, none larger than 5 mm, and located in the same calyx or pelvis as the tip of the nephrostomy tube. PMID- 12775090 TI - Life threatening hematuria in a patient with renal angiomyolipoma and selective renal embolization prior to nephrectomy. PMID- 12775091 TI - Urological symptoms of acute appendicitis in childhood and early adolescence. AB - We present 15 cases of acute appendicitis in ten boys and five girls (age 3-15 years) with cardinal symptomatology coming from the urogenital tract, who were treated in our departments. All the patients presented with right renal colic, dysuria, frequency and urinary retention. The symptoms were attributed to an ongoing appendix inflammatory process in close proximity to the right distal ureter and urinary bladder. All the patients were successfully operated, and postoperative courses were uneventful. As the present patient group is the largest reported to date, a classification of the pathophysiology in relation to the clinical presentation is proposed. PMID- 12775092 TI - Right upper quadrant swelling due to leiomyosarcoma. AB - Reports of leiomyosarcoma occurring in the extremely elderly are quite rare. We describe a case of leiomyosarcoma in an 85-year-old patient and discuss magnetic resonance findings. PMID- 12775093 TI - Haemoperitoneum secondary to ureteric stenting. AB - We report an unusual but catastrophic complication of 'haemo peritoneum' noted following an uneventual insertion of ureteric stent. Its management was primarily affected by the presence of a significant coexisting pathology. This case highlights two important points: (1) recognizing the most rare but real possibility of major intra peritoneal haemorrhage following simple stenting of the ureter (2) the importance of checking that the guide wire has been correctly preloaded in the spool before its placement up the ureter as inserting the wrong (nonfloppy) end may have serious implications. PMID- 12775094 TI - Bacterial colonization of double J stents and bacteriuria frequency. AB - AIM: Stents and catheters are widely used in urology. In our study the frequency of double J stent (DJS) colonization and stent associated bacteriuria is investigated. MATERIALS AND METHODS: Between June 2000-April 2001, 61 patients (17-72 years old) to whom DJ stenting was done (5 of them bilateral) were included in this study. Stone extraction/lithotripsy, extracorporeal shockwave lithotripsy (ESWL), open urolithiazis surgery and hydronephrosis were the indications. Sterile urine samples were obtained prior to stenting. Patients were clinically followed and after the treatment DJ stents were removed after having urinary cultures via the cystoscope. DJS cultures were also made. RESULTS: DJS indwelling times were between 15-90 days (52.5 +/- 8.6). In 3 female, 8 male totally 11 patients bacterial colonizations were observed. In 10 of them both stent and urinary cultures were positive, and showed identical microorganisms; 80% E. Coli, 10% E. Coli+ Candida, 10% Klebsiella. CONCLUSIONS: Our infection rate of DJS and urine is 18% which is not very high. Routine DJS cultures are not needed as the same microorganisms grow in urine cultures. No age or sex relation to the infections is found. Antibiotic prophylaxis is effective in preventing clinically ouvert infectious diseases. PMID- 12775095 TI - A very unusual intravesical foreign body in a male. AB - We present a case of a battery as foreign body in the bladder in a 36-year-old man. Many kinds of foreign bodies in the bladder have been reported but to our knowledge this is the first case of a battery. The diagnosis and the treatment of the case is discussed. PMID- 12775096 TI - Case report: an unusual intravesical foreign body: cause of recurrent urinary tract infections. AB - A 16-year-old boy presented with recurrent urinary tract infections in spite of antibiotic treatment. Investigations revealed the presence of an unusual intravesical foreign body i.e. a common ballpoint. It was removed via cystostomy. In any patient presenting with recurrent urinary tract infections the possibility of an intravesical foreign body should always be considered. PMID- 12775098 TI - Epididymal leiomyoma associated with leydig cell hyperplasia: a case report and immunohistochemical study. AB - Leiomyomas are rare soft tissue neoplasms that can affect virtually any part of the male genital tract. Although they have been described in association with other lesions, to best our knowledge coexistence with leydig cell hyperplasia has not been reported before. We present here the clinical, histopathologic and immunhistochemical characteristics of an epididymal leiomyoma associated with leydig cell hyperplasia and an epididymal cyst in a sixty-eight year old man. PMID- 12775097 TI - Chemoprevention of carcinoma prostate: a review. AB - PURPOSE: Chemoprevention of prostate cancer is the administration of agents to prevent, inhibit, or delay progression of prostate cancer. Opportunities exist for testing various types of chemopreventive interventsion. MATERIAL AND METHODS: The authors reviewed the relevant articles published in the last twenty years and studied the biology of the prostate cancer. An attempt is made to identify intermediate markers and surrogate endpoint markers. The various interventions and initial clinical trial results are described. End points for evaluation are mainly based on changes in PSA, changes of histological precursors, or time of onset of clinical disease. RESULTS: Nutritional factors such as reduced fat intake, vitamin A, vitamin E, vitamin C, vitamin D, Lycopene and selenium may have a protective effect against prostate cancer. CONCLUSION: Numerous studies implicate dietary and nutritional factors in the onset and progression of prostate cancer. Hence, it is possible that bioactive compounds (anti-oxidants) like vits. A, C, D, E, minerals like selenium and carotenoids like lycopene can be a part of chemopreventive strategies for prostate cancer. Ongoing studies on nutrition and prostate cancer may bring the required evidence to support what is still only a hypothesis at present. However, absolute recommendation will have to await the results of long term prospective clinical trials. PMID- 12775099 TI - Chronic epididymitis (epididymal nodule) mimicking an adenomatoid tumor--case report with review of literature. AB - AIM: To describe and review the differential diagnosis of epididymal nodules and chronic epididymitis so as to have a broad view of this pathology. We have suggested a possible diagnostic algorithm for the workup of an epididymal nodule. METHODS/RESULTS: We have reported a case of a 35-year-old patient with a symptomatic palpable nodular mass in the tail of the left epididymis. Fine needle aspiration suggested an adenomatoid tumor while the surgical excision histology was reported as chronic epididymitis. CONCLUSIONS: Epididymal nodules are frequently encountered in the epididymis. Their differential diagnosis includes chronic granulomatous epididymitis, adenomatoid tumor and benign paratesticular neoplasms. We consider that it is important to distinguish epididymal nodules from benign inflammatory lesions and the threshold for a surgical excision should be low as it is therapeutic and provides a definite diagnosis. Whilst the FNAC alone may not be always adequate to confirm diagnosis, an epididymectomy may be curative as well as diagnostic in selected cases. PMID- 12775100 TI - Plaque incision and venous patch grafting for Peyronie's disease. AB - PURPOSE: To evaluate the results of plaque incision and venous patch grafting to correct the dorsal curvature associated with Peyronie's disease. MATERIALS AND METHODS: 18 patients with Peyronie's disease were treated surgically. All of the patients have dorsal penile curvature for more than one year causing intromission impossible or with some difficulty because of curvature. Median penile angulatio was 60 degrees (range 45 degrees-75 degrees). A transverse incision about 2 cm long was made on the plaque where maximal site of curvature was identified. The saphenous vein harvested from the ankle region is opened longitudinally and its endothelial surface is placed and sutured in contact with the tunical defect. RESULTS: Complete penile straightening was achieved in 16 (88.8%) cases and only two patients have residual curvature less than 30 degrees permitting sexual intercourse. None of the patients returned to baseline preoperative angulation state in the follow up period. 3 patients (16.6%) reported penile shortening, but only one was concerned and showed dissatisfaction about it. 6 patients (33.3%) have transient changes in penile sensation that resolved within 4 months. CONCLUSIONS: Plaque incision and placement of saphenous vein patch graft offers a 94.4% satisfactory result in Peyronie's disease causing penile dorsal curvature. PMID- 12775101 TI - Penile epidermoid cyst in an elderly patient. AB - Although cutaneous epidermoid cysts are common lesions, penile localization of them is quite rare especially in elderly ages. A case of 70-year-old man with a rapidly growing penile epidermoid cyst that extended to under the symphysis pubis is reported. The cystic mass was excised totally and histopathologic examination revealed the epidermoid cyst. There has been no finding of recurrence for one year follow-up period. PMID- 12775102 TI - Progression to wet gangrene in penile necrosis and calciphylaxis. AB - We report a patient who presented with gangrene of the penis due to renal failure and calciphylaxis. In view of his advanced underlying conditions we pursued an expectant approach. Superinfection set in necessitating extensive debridement, demasculinisation and a perineal urethrostomy. Of the 34 reported cases in the literature, 12 (35%) were treated conservatively, 18 (53%) had been treated expectantly with delayed surgery following the development of complications and 4 (12%) underwent early surgery (penectomy). 20 (59%) of the patients subsequently died--8/12 (58%) treated conservatively, 11/18 (61%) treated with expectant surgery and 1/4 (25%) treated with early surgery. Progression to wet gangrene developed in the majority of patients--23/34 patients (68%). Penile gangrene is a symptom of severe systemic disease, where two thirds of affected patients decease within six months of onset of symptoms. Expectant treatment results in superinfection and wet gangrene in the majority of cases who ultimately require extensive surgery and debridement. PMID- 12775103 TI - Management of post-traumatic arterial priapism in children: presentation of a case and review of the literature. AB - In this article, a 9-year-old boy with arterial priapism is presented. The patient was managed with the conservative measures including imipramine hydrochloride and a favorable outcome was achieved after 2 months of follow-up. The pathophysiology, diagnostic tools and treatment alternatives are discussed. PMID- 12775104 TI - Use of veno-venous bypass for resection of malignant pheochromocytoma with vena caval thrombus. AB - Surgical management of malignant pheochromocytoma with tumor-induced venous obstruction involving the entrance to the right atrium is challenging. The risk of marked hypotension and hemodynamic instability following clamping of the vena cava is increased as a consequence of the sudden decrease in circulating catecholamines. The use of cardiac bypass, however is burdened with additional operating time and coagulopathy. The present report illustrates that veno-venous bypass is a valuable tool during resection of phenochromocytoma with a large vena caval tumor thrombus. PMID- 12775105 TI - Predictive pathological factors of lymph nodes involvement in the squamous cell carcinoma of the penis. AB - OBJECTIVES: To evaluate the predictive role of primary tumor histopathological features in predicting inguinal lymph nodes involvement in patients with penile squamous cell carcinoma. MATERIAL AND METHODS: We retrospectively analysed pathological records from 30 consecutive patients who underwent penectomy for invasive squamous cell carcinoma of the penis. All histological specimens were reviewed by the same pathologist. We considered the following histological parameters: histological grading, growth pattern, deph invasion, tumour thickness, nuclear grading, poorly differentiated cancer rate, vascular and lymphatic embolization, eosinophilic and mononuclear infiltration and pathological stage. RESULTS: Lymph nodes involvement occurred in 5 patients who underwent 'early' lymphadenectomy and in other 4 ones during oncological surveillance. Lymph nodes metastasis resulted significantly correlated with histological grading (p = 0.005), lymphatic (p = 0.005) and venous (p = 0.02) embolization, corpora cavernosa (p = 0.03) and urethra (p = 0.03) infiltration. Histological grading and lymphatic embolization were independent predictive variables of lymph nodes involvement (p = 0.02). CONCLUSIONS: The histological grading and lymphatic embolization have to be considered as important parameters to select patients with penile squamous cell carcinoma to undergo an 'early' lymphadenectomy. PMID- 12775107 TI - Geriatric nephrology and the 'nephrogeriatric giants'. PMID- 12775106 TI - Kaposi's sarcoma of the glans penis in an HIV negative patient. PMID- 12775108 TI - Diabetic nephropathy-pathophysiology and management. AB - Diabetes mellitus is the leading cause of end-stage renal disease in the United States. Between 1996 and 2001, the prevalence of diabetes in the Medicare population increased by 319%. Patients with diabetes account for approximately one-third of all cases of end-stage renal disease (ESRD). This number is expected to rise dramatically as a result of the growing incidence of diabetes and the aging population. A major complication of diabetes includes end-stage renal disease as a result from diabetic nephropathy. The earliest clinical evidence that nephropathy exists is the appearance of low, yet abnormal, levels of albumin in the urine, referred to as microalbuminuria. This can progress to proteinuria representing overt diabetic nephropathy. Prevention remains the best way to reduce mortality and maintain a high quality of life in these individuals as recent clinical trials confirm that it is possible to not only slow down the progression of diabetic nephropathy, but even prevent it from becoming a significant problem. This article reviews the pathogenesis, diagnostic screening, and treatment strategies of diabetic nephropathy. PMID- 12775109 TI - Conversion of intravenously administered beta-cryptoxanthin to a beta-carotene like compound in mouse lung. AB - The intravenous administration of beta-cryptoxanthin (320 microg/mouse) into 4-wk old ddY mice caused a new peak in addition to the peak of beta-cryptoxanthin during high-performance liquid chromatography (HPLC) of the lipid fraction of lung homogenate. Different HPLC conditions revealed that the new peak might be attributed to a beta-carotene-like compound. The average retention times for the new peak and authentic all-trans-beta-carotene were 14.97 and 15.00 min, respectively, in a HPLC system using a YMC-Pack ODS-A column and methanol-based mobile phase, and 27.05 and 26.93 min, respectively, in a HPLC system using a Waters Nova Pack C18 column and methanol-based mobile phase. In a HPLC system using a Waters Nova Pack C18 column and acetonitrile-based mobile phase, the retention times were 10.73, 10.48 and 10.70 min for the new peak, authentic all trans-beta-carotene, and 9-cis-beta-carotene, respectively. Spectrophotometry with a photodiode array detector showed maximum absorption of 447 and 475 nm for the new peak, and 450 and 475 nm for authentic all-trans-beta-carotene. This new peak was not observed in the lung tissue of control mice. These findings indicate the possible conversion of beta-cryptoxanthin to a beta-carotene like-compound in ddY mice. PMID- 12775110 TI - Biosynthesis of pyridoxine in Saccharomyces cerevisiae--origin of the pyridoxine nitrogen atom differs under anaerobic and aerobic conditions. AB - The amide nitrogen atom of glutamine is incorporated into pyridoxine in four eukaryotes (i.e., Emericella nidulans, Mucor racemosus, Neurospora crassa and Saccharomyces cerevisiae) and two prokaryotes (i.e., Staphylococcus aureus and Bacillus subtilis). However, in the prokaryotes Pseudomonas putida, Enterobacter aerogenes and Escherichia coli, it is the nitrogen atom of glutamate that is incorporated into pyridoxine (J Nutr Sci Vitaminol (2000) 46, 55-57). As these results were from experiments conducted under aerobic conditions, we investigated the biosynthesis of pyridoxine on S. cerevisiae under anaerobic conditions. The results showed that [amide-15N]L-glutamine was not incorporated into pyridoxine, unlike the results for aerobic conditions. The incorporation of [15N]ammonium salts into pyridoxine was not inhibited in the presence of casamino acids and tryptophan. The results showed that the nitrogen atoms of amino acids are not used for the biosynthesis of pyridoxine. The incorporation of 15N into pyridoxine was inhibited in the presence of adenine, but not in that of hypoxanthine. Thus, the nitrogen atom of pyridoxine may be from the amino group attached to the C-6 of adenine. PMID- 12775111 TI - Response of the insulin-like growth factor system to vitamin A depletion and repletion in rats. AB - Vitamin A (VA) and insulin-like growth factors (IGF) are important regulators of a wide range of physiological processes. To investigate the IGF system's involvement in the physiological actions of VA, we examined the effects of VA status on components of the IGF system in rats. Male rats (3-wk-old) fed a VA deficient diet for 11 wk developed VA deficiency, as confirmed by the depletion of serum retinol and hepatic retinyl palmitate. Rats fed the VA-deficient diet had significantly lower body weight (p < 0.05) and lower serum IGF-I concentrations than the rats fed the control diet. The decreases in serum IGF-I levels were accompanied by approximately 40% lower levels of the IGF-I mRNA in the liver and lungs. With respect to the gene expression of other IGF system components, VA deficiency caused a twofold induction of IGF-I receptor (IGF-IR) mRNA in the heart and a twofold reduction in IGFBP-6 mRNA in the lungs, but did not alter the expression of IGF-II, IGFBP-1, IGFBP-3, IGFBP-4 or IGFBP-5 in all tissues examined. When VA-deficient rats received a single injection of retinoic acid (2 mg/rat), tissue IGF-I and IGF-IR gene expression did not change after 4 or 8 h, while the expression of IGF-II, IGFBP-4, and IGFBP-6 mRNAs in some tissues increased rapidly. These results suggest a possible involvement of the IGF system in mediating the physiological actions of VA, including VA-supported growth, in the rat. PMID- 12775112 TI - The effect of dietary iron levels on changes in iron status and zinc-dependent enzyme activities in rats fed two levels of dietary zinc. AB - In order to determine the interrelationship between dietary iron and zinc levels, the effects of dietary iron levels (2, 10, 20, and 40 microg/g) on changes in iron and zinc status and zinc enzyme activities (aminolevulinic acid dehydratase ALA-D EC 4.2.1.24 and alkaline phosphatase ALK-P EC 3.1.3.1) in male Wistar rats were investigated using adequate and marginally deficient zinc diets (25 and 5 microg/g). When rats were fed 5 microg Zn/g diets, body weight gain and food intake remained unchanged at a Fe diet intake of 20 microg/g or greater. Similar tendencies were obtained for hemoglobin, hematocrit, plasma iron, and transferrin saturation. In contrast, liver, spleen, and femur iron concentrations increased gradually with increased iron intake. Feeding diets containing 25 microg Zn/g did not alter these parameters. The percentages of apparent iron absorption in both dietary zinc groups tended to increase with decreasing dietary iron and attained maximum levels at an Fe intake of 10 microg/g. However, In the case of rats fed Fe at concentrations of 2 microg/g Iron absorption decreased. Regardless of the dietary zinc level, rats fed diets with an Fe concentration of 2 microg/g had decreased zinc absorption and plasma ALK-P activity. However, ALA-D activity was not influenced by dietary iron. PMID- 12775113 TI - Validity of a self-administered food frequency questionnaire against 7-day dietary records in four seasons. AB - The validity of a food frequency questionnaire (FFQ) was evaluated against four 7 d diet records among 23 men aged 27-70 y. Volunteer women recorded the amounts of all foods and beverages consumed by their husbands or fathers over a period of 7 d in four consecutive seasons in 1997 and 1998, and a self-administered FFQ was answered by each man after completion of the fourth diet record. The diet record was kept in accordance with the method used for Japan's National Nutrition Survey. The FFQ ascertained frequencies and amounts of consumption of rice, bread, noodles, green tea, and coffee: consumption frequency of 35 food items; and consumption frequency and amount of five alcoholic beverages. As for most nutrients and foods, mean intakes estimated by the FFQ were lower than those estimated from the diet record. Crude intake of nutrients showed fairly good agreement between the two methods in terms of Pearson's correlation coefficient (r > 0.5), except for total energy (r = 0.23), fat (r = 0.36), and vitamin A (r = 0.29). The adjustment for energy intake generally reduced the correlation; the reduction being fairly large for fat intake and vitamin A (r = 0.19 for both). There was generally good agreement for the intake of foods and beverages with exceptions for fats/oils (Spearman correlation coefficient, rs = 0.30) and other vegetables (rs = 0.35). The highest correlation was observed for alcoholic beverages (rs = 0.91), bread (rs = 0.80), and fruits (rs = 0.77). An FFQ covering a limited number of common foods may be useful in assessing the relative position of an individual's habitual consumption of foods and nutrients; however, the instrument generally underestimates absolute intake. PMID- 12775114 TI - Differential effects of vegetable-derived indoles on the induction of quinone reductase in hepatoma cells. AB - The increased expression of quinone reductase (QR) has been associated with anticarcinogenic processes. The aim of this study was to explore the roles of the cruciferous vegetable-derived indoles, indole-3-carbinol (I3C) and indolo[3.2 b]carbazole (ICZ), on the regulation of QR in both murine (Hepa-1) and human (HepG2) hepatoma cells. The results indicate that ICZ enhanced QR activity in both Hepa-1 and HepG2 cells, whereas its parent compound. I3C, had no significant effect on the induction of QR. Moreover, the ICZ-induced QR activity showed a higher response and expressed a more-significant dose-response in Hepa-1 cells. QR mRNA expression as analyzed by RT-PCR demonstrated a pattern similar to that of the enzyme activity. in conclusion, I3C did not show an enhancement effect on OR activity, but its acidic derivative, ICZ, increased the expression of QR mRNA, which then caused the augmentation of QR activity in Hepa-1 and HepG2 cells. PMID- 12775115 TI - The effect of taurine on plasma cholesterol concentration in genetic type 2 diabetic GK rats. AB - The purpose of this study is to investigate the effect of taurine on the plasma cholesterol concentration in genetic type 2 diabetic rats fed cholesterol-free or high-cholesterol diets. Diabetic rats (GK male rats) and normal rats (Wistar male rats) were fed either a cholesterol-free or cholesterol-enriched (1% cholesterol + 0.25% sodium cholate) diet supplemented with or without 3% taurine for 21 or 14 d. Compared to the normal rats, diabetic rats showed a high glucose concentration in their blood and plasma, but it was not affected by taurine feeding. The plasma insulin concentration was higher in the diabetic rats than in the normal rats. At the start of the experiment, the plasma cholesterol concentration was significantly higher in the diabetic rats than in the normal rats. Taurine did not affect the plasma cholesterol level in rats fed the cholesterol-free diet. However, taurine feeding significantly increased the plasma HDL-cholesterol concentration in the diabetic rats fed the cholesterol-free diet. In both the diabetic and normal rats fed the cholesterol diet, the plasma cholesterol concentration was significantly lower in rats fed the diet supplemented with taurine than in the rats fed the control diet. It was concluded that taurine has a hypocholesterolemic effect in both diabetic and normal rats fed diets containing cholesterol. Moreover, these results suggest that taurine seems to affect the HDL-cholesterol metabolism in diabetic rats fed a cholesterol-free diet. PMID- 12775116 TI - Beneficial effects of different Polygonum multiflorum Thunb. extracts on memory and hippocampus morphology. AB - Four groups of 1-mo-old male senescence-accelerated mice (SAMP8) were fed Polygonum multiflorum Thunb. (PM) extract for 18 wk to determine the effect of PM on memory ability and histopathological changes in mice. The baseline diet consisted of a casein diet group, and the three test diets were supplemented with 50% ethanol, 95% ethanol, or water extracts of PM. It was found that the mice fed with PM extracts had better active shuttle avoidance response, fewer vacuole numbers, less lipofuscin in the hippocampus, and lower MDA concentrations in the brain. Our data showed that the ethanol PM extract groups (both 50% and 95% groups) had lower lipofuscin percentages and MDA concentrations, and higher total thiol concentrations than the water PM extract group. The 50% ethanol PM extract group showed significantly lower total cholesterol and triglyceride values than the other groups, but the HDL cholesterol level was the same. These results suggest that dietary supplementation with either ethanol or water extracts can reduce brain pathological changes and promote learning and memory ability. The performance of PM extracts depended on the extraction method, with ethanol extraction tending to obtain better results. PMID- 12775117 TI - Validity and reliability of a dietary assessment method: the application of a digital camera with a mobile phone card attachment. AB - This study was aimed at evaluation of the validity and reliability of an alternative dietary measurement method that assists epidemiologic studies. We validated a handheld personal digital assistant with camera and mobile phone card, called Wellnavi, in which a 1-d weighed diet record was employed as a reference method. Twenty college students majoring in food and nutrition participated in this study. They were asked to keep a diet record and to take digital photos of all these recorded food at the same time, then send them to the dietitians by the mobile phone card. In the reliability study, other twenty students from the same college were asked to take digital photos of the same meal during a day by two same instruments under the same circumstances and to send these photos to the different dietitians electronically. With respect to validity, median nutrient intakes estimated by the Wellnavi method and the diet record method are comparable. Correlation coefficients between the median nutrient intakes estimated from these two methods ranged from 0.46 for monounsaturated fatty acid to 0.93 for vitamin B12 and copper (median r = 0.77). With respect to reliability, our data show a good agreement between two Wellnavi instruments for most of the nutrients. Correlation coefficients between the nutrient intakes estimated from 2 instruments ranged from 0.55 for vitamin B1 and water-insoluble dietary fiber to 0.92 for vitamin B12 (median r = 0.78). In conclusion, the results indicate this dietary assessment instrument can usefully measure individual dietary intakes for a variety of nutrients in an epidemiologic study. PMID- 12775118 TI - Dose-dependent effect of dietary conjugated linoleic acid on the growth of rat hepatoma dRLh-84 cells in vivo. AB - In this study, the effect of varying doses of conjugated linoleic acid (CLA) on the growth of transplanted hepatoma dRLh-84 cells and the relationship between tumor growth and prostaglandin (PG) E2 production or cyclooxygenase (COX)-2 expression were examined. Donryu rats were fed an experimental diet containing 0, 0.1, 0.5, or 2 wt.% CLA for 3 wk, and then dRLh-84 cells were transplanted into the liver. Results show that dietary CLA (0.5 and 2 wt.%) significantly enhanced the growth of the transplanted hepatoma cells compared to the non-CLA diet group at 20 d after cell transplantation. Tumor weight at 10 d after transplantation was also significantly higher in the 2 wt.% CLA group than in non-CLA fed rats. Ten days after transplantation, the PGE2 level in the tumor tissue was shown to be depressed in a CLA dose-dependent manner. Cyclooxygenase-2 (COX-2) mRNA expression in the tumor also tended to be lower in the CLA group than in the non CLA diet group 10 d after transplantation. Dietary CLA did not affect the tumor phospholipid arachidonic acid level, which is a substrate for PG synthesis. These results indicate that dietary CLA of at least 0.5 wt.% enhances the growth of transplanted dRLh-84 cells in vivo. It is believed that growth promotion of dRLh 84 cells in vivo by CLA cannot be clarified by the PG synthesis dependent mechanism. PMID- 12775119 TI - Effects of oral acute administration and subchronic feeding of several levels of D-psicose in rats. AB - The effects of oral acute administration and subchronic (34 d) feeding of several levels of D-psicose, a C3-epimer of D-fructose, were studied in rats. In the acute administration test, five groups of eight male Wistar rats (3 wk old) were orally given D-psicose in doses of 8, 11, 14, 17, and 20 g/kg. Three rats receiving 14 g/kg, three rats receiving 17 g/kg and eight rats receiving 20 g/kg of D-psicose died within 2 d after administration. The calculated LD50 values were 16.3 g/kg by the Behrens-Karber method and 15.8 g/kg by the Litchfield Wilcoxon method. In the subcronic feeding test, eight groups of seven male Wistar rats (3 wk old) were fed diets containing 0 (control), 10, 20, 30, and 40% for 34 d. One rat fed 30% D-psicose diet and five rats fed 40% D-psicose diet died during the experimental period. Body weight gain, food intake and food efficiency were more extensively suppressed by the higher D-psicose diets. The weights of heart, spleen and abdominal adipose tissue were smaller in the order of dietary D psicose concentration. Cecal weight increased with increasing D-psicose concentration in the diets. Cecal hypertrophy was observed in rats fed 10-40% D psicose diets. These results suggest that D-psicose differs in nutritional characteristics from D-glucose or D-fructose. The feeding of diets extremely high in D-psicose seems to be harmful to the intestinal tract. PMID- 12775120 TI - Larger diet-induced thermogenesis and less body fat accumulation in rats fed medium-chain triacylglycerols than in those fed long-chain triacylglycerols. AB - It has been previously shown that a diet containing medium-chain triacylglycerols (MCT) leads to less body fat accumulation as compared to a diet containing long chain triacylglycerols (LCT). We investigated the involvement of diet-induced thermogenesis in the accumulation of body fat in rats fed a diet containing MCT. Twelve male Wistar rats were administered 1 g of MCT or LCT by gavage, and their oxygen consumption was measured for 6 h (experiment 1). Forty male Wistar rats were fed a diet containing 10% MCT or LCT for 6 wk, and their body composition was determined (experiment 2). In experiment 1, oxygen consumption increased to a greater extent after MCT administration than after LCT administration. Diet induced thermogenesis was significantly (0.67 +/- 0.14 kcal) larger after the administration of 1 g of MCT. In experiment 2, there were no differences in food intake or carcass protein content between the LCT group and MCT group. However, carcass fat and intra-abdominal fat content were significantly lower in rats fed MCT than in those fed LCT. We calculated that ingestion of 1 g of MCT decreased body fat by 0.94 +/- 0.27 kcal relative to the ingestion of LCT. These results suggest that the larger diet-induced thermogenesis observed in rats fed MCT, compared to that of those fed LCT, is one of the main factors involved in the suppression of body fat accumulation in rats fed MCT. PMID- 12775121 TI - Anthocyanin administration elevates plasma homocysteine in rats. AB - The data accumulated from epidemiological studies suggests that individuals with elevated blood levels of homocysteine have an increased risk of cardiovascular disease. However, little is known of the food factor that may affect the homocysteine status, except for folate and B-vitamins. Here, we tested the effect of dietary phenolics (i.e., anthocyanin of food colorant) administration on plasma homocysteine concentration in a rat study, since a profound effect on the methionine metabolism was speculated from the 3',4'-catechol skeletal structure of anthocyanin. Female Sprague-Dawley rats (body weight 100 g) orally ingested a single dose of anthocyanin mixture (total 100 mg) composed of cyanidin-3 glucoside (50mg), cyanidin-3-sambubioside (48 mg), and cyanidin-3,5-diglucoside (2 mg). The total homocysteine in the plasma collected 90-240 min after anthocyanin intake was 1.4 to 1.8-fold (5.2-6.7 micromol/L) higher than the basal homocysteine level (3.7 micromol/L). In the liver and kidney, anthocyanin significantly affects sulfur amino acid (S-adenosylmethionine, SAM, and S adenosylhomocysteine, SAH) levels, both of which are precursors of plasma homocysteine, and the SAH/SAM ratio showed a significant increase in the liver and kidney. Accordingly, these results suggest that dietary anthocyanin stimulates homocysteine synthesis from SAH in the liver and kidney, and the homocysteine yielded transfers into the blood stream. The intake of anthocyanin and its structural homologues may have an effect on the metabolic regulation of sulfur amino acids and possibly increase the risk of vascular disease in humans. PMID- 12775122 TI - Comparison of diet-induced thermogenesis of foods containing medium- versus long chain triacylglycerols. AB - The purpose of this study was to investigate the effect of 5-10 g of medium-chain triacylglycerols (MCT) on diet-induced thermogenesis in healthy humans. The study compared diet-induced thermogenesis after ingestion of test foods containing MCT and long-chain triacylglycerols (LCT), using a double-blind, crossover design. Eight male and eight female subjects participated in study 1 and study 2, respectively. In both studies, the LCT was a blend of rapeseed oil and soybean oil. In study 1, the liquid meals contained 10 g MCT (10M), a mixture of 5 g MCT and 5 g LCT (5M5L), and 10 g LCT (10L). In study 2, the subjects were given a meal (sandwich and clear soup) with the mayonnaise or margarine containing 5 g of MCT or LCT. Postprandial energy expenditure was measured by indirect calorimetry before and during the 6 h after ingestion of the test meals. Diet-induced thermogenesis was significantly greater after 5M5L and 10M Ingestion as compared to 10L ingestion. Ingestion of the mayonnaise or margarine containing 5 g MCT caused significantly larger diet-induced thermogenesis as compared to that of LCT. These results suggest that, in healthy humans, the intake of 5-10 g of MCT causes larger diet-induced thermogenesis than that of LCT, irrespective of the form of meal containing the MCT. PMID- 12775123 TI - Red mulberry. PMID- 12775124 TI - Four pilot projects completed for school-based allergy and asthma screening. PMID- 12775125 TI - Fungus research can grow on you. PMID- 12775126 TI - Do leukotriene receptor antagonists have a place in pharmacotherapy of allergic rhinitis? PMID- 12775127 TI - Autoimmune progesterone dermatitis and its manifestation as anaphylaxis: a case report and literature review. AB - OBJECTIVE: Autoimmune progesterone dermatitis is a rare cyclic premenstrual reaction to progesterone produced during the luteal phase of a woman's menstrual cycle with a variety of presentations including erythema multiforme, eczema, urticaria, angioedema, and progesterone-induced anaphylaxis. We present a case of progesterone-induced anaphylaxis and a review of literature focusing on its diagnosis and therapy. DATA SOURCES: We surveyed all the literature in English back to 1921 when the first case was published. First, we researched the terms progesterone anaphylaxis, autoimmune progesterone dermatitis, cyclic urticaria, using the PubMed resource. Then we included articles found within these publications' reference sections. STUDY SELECTION: We selected articles based on whether the cases described appeared to fit the description of the entity autoimmune progesterone dermatitis. All cases included had dermatologic reactions occurring during the luteal phase of the menstrual period, positive skin or intramuscular reactions to progesterone, and treatment amenable to anovulatory agents and/or hysterectomy with bilateral salpingo-oophorectomy. RESULTS: We found approximately 50 published cases of autoimmune progesterone dermatitis, and only nine known cases of its manifestation as anaphylaxis. These cases, including the case described by us, are summarized, and successful diagnostic and therapeutic approaches in the literature are reviewed. CONCLUSIONS: Autoimmune progesterone dermatitis is a rare entity associated with progesterone production of the luteal phase of a woman's menstrual cycle. It can be diagnosed using intradermal or intramuscular progesterone tests and can be treated by disrupting the ovulation cycle using specific medications or by oopherectomy. PMID- 12775128 TI - A 12-year-old girl with asthma, pneumonia, and pneumatoceles. PMID- 12775129 TI - Fever, abdominal pain, and interstitial pneumonia in a 13-year-old female. PMID- 12775130 TI - Results of an inner-city school-based asthma and allergy screening pilot study: a combined approach using written questionnaires and step testing. AB - BACKGROUND: A questionnaire alone may not be an adequate screening tool for asthma. OBJECTIVE: To determine whether an asthma questionnaire used in combination with an exercise step test is better than a questionnaire alone in screening for asthma in children and to evaluate the validity of a rhinitis questionnaire in determining atopy. METHODS: The International Study of Asthma and Allergies in Childhood (ISAAC) asthma core questionnaire was used to screen for asthma in 307 inner-city first through third graders. All children who had scores consistent with a diagnosis of asthma underwent step testing, as did a subset of children who had negative overall scores. All children who had inconsistent asthma scores and step test results underwent methacholine challenge testing. The same 307 children underwent rhinitis screening and children who had one or more positive responses on the ISAAC rhinitis questionnaire underwent skin testing as did a subset of children who had all negative responses. RESULTS: Three hundred of 307 asthma and rhinitis questionnaires were returned. Twenty eight children (9%) had global asthma scores that were considered to be positive (5 or above). Twenty-four of these children underwent step testing as did 34 randomly selected children who had negative global asthma scores. Thirty-one (91%) of the 34 children who had negative global asthma scores had negative step tests. Similarly, 20 of 24 children (83%) of the children who had positive global asthma scores had negative step tests. Only 4 children who had positive global asthma scores were step test-positive or had reversible airway obstruction at baseline. Using a positive methacholine challenge as the gold standard for establishing bronchial hyperresponsiveness, the global asthma score derived from the eight-item ISAAC asthma questionnaire yielded a sensitivity of 64%, a specificity of 11%, a positive predictive value of 47%, and a negative predictive value of 20%. Comparing the six-item ISAAC rhinitis questionnaire results to the gold standard, skin test reactivity, the questionnaire yielded a sensitivity of 76%, a specificity of 21%, a positive predictive value of 56%, and a negative predictive value of 40%. CONCLUSIONS: Step testing was not useful as a screening tool for asthma. In addition, the ISAAC asthma questionnaire may not be a good asthma screening tool for inner-city pediatric populations, especially if the form is self-administered. Investigators should first validate both the ISAAC asthma and rhinitis screening questionnaires in the particular population to be studied before widespread asthma and allergy screening efforts are initiated using these tools. PMID- 12775131 TI - Development and validation of a brief pediatric screen for asthma and allergies among children. AB - BACKGROUND: Asthma is the most common disease of childhood, but the recognition and detection remain poor, especially among schoolchildren. There has been an increase in the number of instruments available to detect the risk of asthma earlier in children. We have previously validated a simple, self-reported screen, the Brief Pediatric Asthma Screen (BPAS). OBJECTIVE: To develop a new screen for asthma and allergies based on the BPAS (BPAS+) with the intent of keeping the screen brief and simple, while including allergy detection. METHODS: Questions from the BPAS were extensively revised, and questions regarding allergic rhinitis were added. A panel of parents of asthmatic children reviewed and critiqued the questions. The final BPAS + was distributed in elementary schools, and a cohort of 129 participated in a validation against the gold standard of evaluation by an expert in asthma. RESULTS: For asthma the best items were wheeze, persistent cough, night cough, and response to change in air temperature. The simplest scoring, any 1 of the 4 items, yielded the best balance of specificity (73.6%) and sensitivity (73.3%). For allergy, using all six items, having any one or any two of the items had sensitivity of 71.4% and specificity of 77.3%. CONCLUSIONS: The BPAS+ provides a rapid and valid method for the detection of potential allergy and asthma in schoolchildren. Sensitivity and specificity are acceptable for both asthma and allergies. PMID- 12775132 TI - Outcome results of a school-based screening program for undertreated asthma. AB - BACKGROUND: Undertreatment of asthma is associated with significant potentially preventable morbidity, including frequent school absences. Guideline dissemination and clinician education have met with variable success. School based identification of children with potentially undertreated asthma may provide an alternative strategy for improving asthma management in children. OBJECTIVE: To evaluate the effectiveness of school-based identification of potentially undertreated asthma. METHODS: A controlled trial of school-based identification of children with known but symptomatic asthma using mailed parent surveys, letters recommending medical follow-up, and medical record review to evaluate changes in asthma treatment after referral. RESULTS: Most parents (79.9%, n = 5,116 respondents) responded to the survey and 19.4% (n = 994) of children were reported to have a physician diagnosis of asthma or reactive airway disease. Letters of referral were sent to 489 children with parent-reported asthma who were identified as having potentially undertreated asthma. Approximately one third (31.2%, n = 153) of these children had physician visits, and 92 (18.8% of all referred) had documented medication changes. In addition, there were 20 new physician diagnoses in this group of children. In the control group of 604 children with asthma, there were significantly fewer children with asthma-related visits (131, 21.7%, P = 0.0004) and children with medication changes (74, 12.3%, P = 0.002) in a comparable 6-month window. CONCLUSIONS: School-based screening or case identification increased the number of physician asthma-related visits and changes in asthma therapy. PMID- 12775133 TI - Development and validation of school-based asthma and allergy screening instruments for parents and students. AB - BACKGROUND: The increasing morbidity attributable to asthma among school-aged children suggests the potential utility of school-based asthma screening programs. OBJECTIVE: We report our efforts to develop and validate culturally sensitive and clinically useful screening questionnaires (parent and child versions) for asthma and allergies among urban US school children. METHODS: Instrument development was accomplished through literature review, expert medical and child developmental input, focus group feedback, and a rigorous trial of the instruments in a public school setting. Questionnaires were distributed to 2,800 children and their families in an urban public school system (grades kindergarten through 6). Validity was evaluated by blinded comparison of results against a standardized clinical evaluation in 107 children, with final designations determined by an expert panel. RESULTS: Questionnaires pertaining to 2,083 children were returned (participation rate of 74%). A moderate level of agreement was observed between parent and student questionnaire responses (r values = 0.36 to 0.50; P values < 0.001). The highest frequency of asthma-like symptoms was reported for African-American boys and the lowest for Caucasian girls. The items from the parent questionnaire that best predicted asthma were "breathing problems" (occurring rarely or more; odds ratio 12.8; 95% confidence interval, 4.5 to 36.1) and "problems coughing" (sometimes or more; odds ratio 9.7; 95% confidence interval, 3.6 to 26.5). Considering the presence of cough (sometimes or more) and/or breathing problem (rarely or more) yielded a sensitivity of 80%; a specificity of 75%, a positive predictive value of 50%, and a negative predictive value of 92%. Similar levels of prediction were observed for the items "trouble breathing" and "noisy breathing" as directly reported by the students. Allergic rhinitis was best predicted by report of a runny/stuffy no se (sometimes or more; sensitivity of 83%, specificity of 61%). Allergic conjunctivitis was best predicted by "itchy eyes." CONCLUSIONS: Administration of a school-based questionnaire is feasible, with a high response rate and excellent internal consistency. A high sensitivity and acceptable specificity was achieved by using one to two questions for asthma, allergic rhinitis, and allergic conjunctivitis. Among the children in grades 2 or above, comparable levels of prediction could be achieved with the student or parent version. PMID- 12775134 TI - A sensitive two-site enzyme-linked immunosorbent assay for measurement of the major Alternaria alternata allergen Alt a 1. AB - BACKGROUND: Alt a 1 is the major allergen in Alternaria alternata, one of the most important fungi associated with allergic diseases. Mold allergenic extracts show considerable heterogeneity, and thus accurate standardization of these extracts is essential to guarantee their quality. OBJECTIVE: To develop an Alt a 1-specific assay and to evaluate the correlation of Alt a 1 content with the IgE binding activity of A. alternata extracts. METHODS: Recombinant Alt a 1 was produced as nonfusion protein from a polymerase chain reaction-cloned complementary DNA Alt a 1 sequence. Natural Alt a 1 was purified from spent culture medium. Monoclonal and polyclonal antibodies directed to Alt a 1 were produced and used to construct a specific Alt a 1 enzyme-linked immunosorbent assay (ELISA). RESULTS: The ELISA developed was highly reproducible and sensitive, with a detection limit lower than 0.5 ng/mL and a practical working range of 0.5 to 50 ng/mL. The assay was able to detect an Alt a 1-like protein in Stemphylium extracts. Identical parallel dose-response curves were observed when natural Alt a 1 and recombinant Alt a 1 were used as standard. A good correlation was obtained between Alt a 1 content of 13 A. alternata extracts and their IgE binding activity. Alt a 1 was responsible for 70% of the IgE-binding activity of the whole extract. CONCLUSIONS: This sensitive and specific Alt a 1 assay allows the quantification of this major mold allergen and represents a useful tool for the standardization of A. alternata extracts in mass units. It also provides a reliable indication of the allergenic activity of the whole extract. PMID- 12775135 TI - Fluticasone propionate aqueous nasal spray provided significantly greater improvement in daytime and nighttime nasal symptoms of seasonal allergic rhinitis compared with montelukast. AB - BACKGROUND: The safety and efficacy of intranasal corticosteroids for the treatment of allergic rhinitis is well documented in the literature. Additionally, an expert panel has concluded that intranasal corticosteroids are the first line of therapy when obstruction is a major component of rhinitis. Montelukast is a leukotriene receptor antagonist recently approved for the treatment of seasonal allergic rhinitis (SAR). OBJECTIVE: This randomized, double blind, double-dummy, parallel-group study was conducted to compare the effectiveness of a 15-day course of intranasal fluticasone propionate 200 microg, once daily (FP200QD), to oral montelukast 10 mg, once daily (MON10QD), in relieving daytime and nighttime nasal symptoms associated with SAR. METHODS: The intent-to-treat (ITT) analysis population consisted of 705 eligible males and females (> or = 15 years) with SAR randomized to either FP200QD (N = 353) or MON10QD (N = 352). The primary efficacy endpoint was the mean change from baseline in subject-rated daytime total nasal symptom scores (the sum of four individual scores: nasal congestion, itching, rhinorrhea, and sneezing), evaluated via visual analog scales, and averaged over weeks 1 to 2. Secondary endpoints included the four daytime individual nasal symptom scores, the nighttime total, and individual nasal symptom scores (each evaluated on a four point scale from 0 to 3). RESULTS: Statistically significant differences favoring FP200QD over MON10QD were observed for the mean change from baseline in daytime total nasal symptom scores (P < 0.001), daytime individual nasal symptom scores (P < 0.001), nighttime total (P < 0.001), and all individual nasal symptom scores (P < or = 0.002) over the 15-day treatment period. FP200QD and MON10QD were both well tolerated. CONCLUSIONS: The results of this well controlled study demonstrated that FP200QD was consistently superior to MON10QD with regard to every efficacy endpoint evaluated, including daytime and nighttime nasal congestion, in subjects with SAR. PMID- 12775136 TI - "Real-world" effectiveness of daily controller medicine in children with mild persistent asthma. AB - BACKGROUND: Unmeasured confounders and selection bias can significantly influence the results of retrospective observational analyses of asthma therapy. OBJECTIVE: To evaluate the efficacy of oral montelukast and inhaled fluticasone propionate in a randomized, prospective 12-month "real-world" observational analysis of children with mild persistent asthma. METHODS: Children (n = 104) between 6 and 15 years of age with mild asthma as determined by forced expiratory volume in 1 second, symptoms, and evaluation by an experienced pediatric allergist or pulmonologist, who were not currently receiving controller therapy, were randomly assigned to fluticasone or montelukast on an alternating basis. Subjects were asked to complete a questionnaire at 6 and 12 months; otherwise, medical care was identical to that of similar managed care patients. Outcome parameters were evaluated after 12 months by claims database analysis. An acute asthma attack requiring emergent care was the primary outcome parameter. Measures of adherence, symptoms, and asthma control, as measured by the pediatric Asthma Therapy Assessment Questionnaire, were secondary outcome parameters. RESULTS: Demographics, spirometry, symptoms at enrollment, emergent care visits, asthma hospitalizations, routine office visits, and symptoms at study completion were not significantly different between study groups. Adherence, as evaluated by the number of controller fills, was significantly (P = 0.0003) better for montelukast (7.65 +/- 3.01) than fluticasone (5.46 +/- 3.01). Similar numbers of subjects in each study group required beta-agonists and oral prednisone. CONCLUSIONS: These results suggest that oral montelukast and inhaled fluticasone have similar real world efficacies in the treatment of children with mild asthma, possibly as a result of the significantly better adherence with oral montelukast therapy compared with inhaled fluticasone. PMID- 12775137 TI - Providing dust mite-proof covers improves adherence to dust mite control measures in children with mite allergy and asthma. AB - INTRODUCTION: The most critical dust mite avoidance technique for asthmatic children with mite allergy is the use of dust mite-proof bedding covers. Adherence to allergen control measures is poor due to cost. The purpose of this study was to determine whether giving families mite-proof bedding encasings at the time of the diagnosis with mite allergy would improve adherence at a home visit 2 months later. METHODS: Parents of children (mean age 7.7 years) with asthma and positive skin tests for dust mite were assigned either to an intervention group, which received dust mite covers upon enrollment or a comparison group, which did not. Both groups were instructed in dust mite control measures by a medical provider and told to invest in the covers as part of standard care. Two months after enrollment, parent report and observational measures from each family were obtained during a home visit. RESULTS: The group provided allergen encasings was found to be significantly more likely to have used the casings at the home visit, t(22) = 2.77, P < 0.05. There were no differences between groups in adherence to other mite control strategies. Parenting stress was significantly related to poorer mite control compliance. The most common reason reported for nonadherence to control measures was cost. CONCLUSIONS: Providing mite bedding encasings at the time of diagnosis significantly increases compliance at a 2-month home visit. Medicaid and insurance companies should be encouraged to pay for casings as a cost effective measure to improve asthma care in children with mite allergy. PMID- 12775138 TI - Effects of fexofenadine, diphenhydramine, and placebo on performance of the test of variables of attention (TOVA). AB - OBJECTIVE: To determine the effects of diphenhydramine 50 mg and fexofenadine 180 mg on cognitive performance using the Test of Variables of Attention (TOVA), and to ascertain whether the TOVA was sufficiently sensitive to differentiate between the effects of these first- and second-generation H1-receptor antagonists on performance. STUDY DESIGN: The study used a double-blind, placebo-controlled, randomized, crossover design. Forty-two subjects completed four separate TOVA tests: at baseline and after administration of placebo, diphenhydramine 50 mg, and fexofenadine 180 mg. On each occasion, subjects rated subjective feelings of drowsiness on a visual analog scale (VAS) before taking the TOVA. RESULTS: Compared with placebo, diphenhydramine caused an increased response time (P = 0.0230) and more omission errors (P = 0.0398). Diphenhydramine was also associated with increased drowsiness VAS ratings (P = 0.0065) compared with placebo. Diphenhydramine caused significantly more commission errors than fexofenadine (P = 0.0354). Neither placebonor fexofenadine 180 mg caused significant changes in any TOVA or VAS measurements compared with baseline. Fexofenadine was not statistically different from placebo for any evaluation. CONCLUSIONS: The TOVA was sufficiently sensitive to differentiate between the central nervous system effects of fexofenadine and diphenhydramine. Fexofenadine 180 mg had no significant effect on the TOVA measures of performance or on self reported drowsiness compared with placebo. In contrast, diphenhydramine 50 mg caused significant increases in omission errors and response time on the TOVA and increases in self-reported drowsiness compared with placebo [corrected]. PMID- 12775139 TI - Determinants of airway hyperresponsiveness in mild asthma. AB - BACKGROUND: Patients with mild asthma may have coexisting severe airway hyperresponsiveness (AHR), although the reasons for this are uncertain. OBJECTIVE: To evaluate the factors that determine AHR in mild asthma. METHODS: We performed a retrospective database evaluation of two groups of patients with mild asthma with forced expiratory volume in 1 second (FEV1) of 80% or more than predicted. Group A (n = 92; mean inhaled corticosteroid dose, 491 microg) had moderate-to-severe AHR to methacholine (provocative dose causing a 20% decrease in FEV1 [methacholine PD20], < or = 100 microg), whereas group B (n = 92; mean inhaled corticosteroid dose, 509 microg) had borderline AHR (methacholine PD20, > or = 800 microg). Both groups were matched for age, sex, inhaled corticosteroid use, and FEV1. RESULTS: From our database, we found 361 patients with an FEV1 of 80% or more than predicted of whom 123 (34%) had a methacholine PD20 of 100 microg or less and 138 (38%) had a methacholine PD20 of 800 microg or more. The methacholine PD20 geometric means (geometric SE) of groups A and B were 25 microg (3 microg) and 5,392 microg (295 microg), respectively. Despite matched mean values for FEV1, compared with group B, group A had a lower predicted forced expiratory flow between 25% and 75% (71% vs 81%, P = 0.007). A greater proportion of group A compared with group B patients were sensitized to house-dust mite (76% vs 54%, P = 0.002). No significant differences were found between groups in terms of presence of rhinitis and sensitization to other individual aeroallergens. CONCLUSIONS: Increased sensitization to house-dust mite and reduced small airway caliber were associated with moderate-to-severe AHR in mild asthma. Skin prick testing to common aeroallergens, especially house-dust mite, should be a routine part in the evaluation of asthmatic patients, including those patients with mild disease. PMID- 12775140 TI - The relationship of housing and household characteristics to the indoor concentrations of Der f 1, Der p 1, and Fel d 1 measured in dust and air samples. AB - BACKGROUND: Indoor dust mite and cat allergens have been related to the risk of atopic conditions. If allergen levels are influenced by modifiable residential characteristics, potential interventions to prevent disease could be deployed. OBJECTIVE: To evaluate relationships between allergen concentrations in air and dust samples and selected house and household characteristics using a large prospective study with multiple sequential allergen measurements from each residence. METHODS: Fel d 1, Der f 1, and Der p 1 were measured in paired air and dust samples collected at intervals throughout 4 years in suburban homes. House and household characteristics were examined for relationships to allergen concentrations in both univariate and multiple variate analyses. RESULTS: The relationships between house and household characteristics and allergen concentrations in both air and dust were complex. When the housing variables were considered in multiple variate analysis, concentrations of Der f 1 in dust increased with increasing number of residents and relative humidity and declined when forced air heating was used. Dust concentrations of Der p 1 were lower in new homes and during forced air heating use but higher with higher relative humidity and in the presence of dogs. The presence of cats was the dominant determinant of Fel d 1 in both air and dust, but when homes without cats were analyzed separately, dust levels of Fel d 1 were inversely related with relative humidity. CONCLUSIONS: Air and dust concentrations of Der p 1 and Der f 1 were positively related to relative humidity and the size of the family. Fel d 1 was positively related to the presence of cats. The relationship of other house or household characteristics was inconsistent but different for Der f 1 and Der p 1. PMID- 12775141 TI - Drug-induced rash with eosinophilia and systemic symptoms syndrome with bupropion administration. AB - BACKGROUND: Sustained-release bupropion is commonly used for the symptomatic relief of depressive illness and as an adjuvant in smoking cessation therapy. OBJECTIVE: To report a case of bupropion-induced drug rash with eosinophilia and systemic symptoms syndrome, including acute hepatitis, obstructive lung disease, and myositis. METHODS: After the patient discontinued use of bupropion, serologic tests, muscle biopsies, pulmonary function tests, a chest x-ray examination, venous Doppler ultrasounds, and an electrocardiogram were performed. RESULTS: On discontinuation of bupropion and prolonged systemic corticosteroid therapy, there was complete resolution of symptoms. CONCLUSIONS: To our knowledge, this is the first reported case of drug rash with eosinophilia and systemic symptoms syndrome induced by bupropion therapy. We report this case to notify clinicians of the potential serious multisystem complications that can occur with sustained-release bupropion therapy. PMID- 12775142 TI - Patient preferences and sensory comparisons of three intranasal corticosteroids for the treatment of allergic rhinitis. PMID- 12775143 TI - Tryptamine-induced resistance in tryptophan decarboxylase transgenic poplar and tobacco plants against their specific herbivores. AB - The presence of amines and their derivatives in plant tissues is known to influence insect feeding and reproduction. The enzyme tryptophan decarboxylase (TDC) catalyzes the decarboxylation of tryptophan to tryptamine, which is both a bioactive amine and a precursor of other indole derivatives. Transgenic poplar and tobacco plants ectopically expressing TDC1 accumulated elevated levels of tryptamine without affecting plant growth and development. This accumulation was consistently associated with adverse effects on feeding behavior and physiology of Malacosoma disstria Hub. (forest tent caterpillar, FTC) and Manduca sexta L. (tobacco hornworm, THW). Behavior studies with FTC and THW larvae showed that acceptability of the leaf tissue to larvae was inversely related to foliar tryptamine levels. Physiological studies with FTC and THW larvae showed that consumption of leaf tissue from the transgenic lines is deleterious to larvae growth, apparently due to a postingestive mechanism. Thus, ectopic expression of TDC1 can allow sufficient tryptamine to accumulate in poplar and tobacco leaf tissue to suppress significantly the growth of insect pests that normally feed on these plants. PMID- 12775144 TI - Antifeedant effects of proteinase inhibitors on feeding behaviors of adult western corn rootworm (Diabrotica virgifera virgifera). AB - Low-molecular-weight peptidyl proteinase inhibitors (PIs) including leupeptin, calpain inhibitor I, and calpeptin were found to be potent antifeedants for adult western corn rootworm (WCR) against the phagostimulation of cucurbitacin B (Cuc B) or a corn pollen extract (CPE). Leupeptin was the strongest (ED50 = 0.36 and 0.55 nmol/disk for Cuc B and CPE, respectively) among PIs tested with an antifeedant potency much stronger than the steroid progesterone (ED50 = 2.29 and 5.05 nmol/disk for Cuc B and CPE, respectively), but slightly less than the reference alkaloid, strychnine (ED50 = 0.17 and 0.37 nmol/disk for Cuc B and CPE, respectively). All active PIs contain a di- or tripeptidyl aldehyde moiety, indicating that PIs exert their antifeedant effects by covalent interaction with putative sulfhydryl (SH) groups on taste receptors as do these PIs with cysteine proteinases. However, opposite inhibition potency against Cuc B versus CPE by two thiol-group reducing agents, DTT and L-cysteine, and the results with other cysteine-modifying reagents obscure the net functional role of SH groups at WCR taste chemoreceptors. Surprisingly, the model phagostimulant for diabroticites, Cuc B, was more easily counteracted by these feeding deterrents than the stimulants present in CPE. Three-dimensional structure-antifeedant relationships for the PIs suggest that a novel taste chemoreception mechanism exists for these peptidyl aldehydes or that they fit partially into a strychnine binding pocket on protein chemoreceptors. Favorable economic benefit may be achieved if PIs are discovered to be useful in adult WCR control, since both pre- and postingestive sites would be targeted. PMID- 12775145 TI - Phenolic compounds on the pod-surface of pigeonpea, Cajanus cajan, mediate feeding behavior of Helicoverpa armigera larvae. AB - A methanol extract of the pod surfaces of Cajanus cajan, a feeding stimulant for fifth-instar Helicoverpa armigera, was shown to contain four main phenolic compounds. Three of these were identified as isoquercitrin, quercetin, and quercetin-3-methyl ether, by comparing UV spectra and HPLC retention times with authentic standards. The fourth compound was isolated by semipreparative HPLC and determined to be 3-hydroxy-4-prenyl-5-methoxystilbene-2-carboxylic acid (stilbene) by NMR spectroscopy and mass spectrometry. Quercetin, isoquercitrin, and quercetin-3-methyl did not affect the selection-behavior of fifth-instar H. armigera. However, larvae were deterred from feeding on glass-fiber disks impregnated with the stilbene. Furthermore, larvae exposed to quercetin-3-methyl ether consumed significant amounts of both disks. In a binary-choice bioassay, a combination of quercetin-3-methyl ether and the stilbene on one disk and pure quercetin-3-methyl ether on the other disk resulted in increased consumption of both glass-fiber disks by larvae. In contrast, consumption was reduced if the combination was presented to larvae on one disk with purified stilbene on the other disk. Cajanus cajan cultivars that varied in their susceptibility to H. armigera were surveyed forthe presence of the four phenolic compounds. An absence of quercetin and higher concentrations of isoquercitrin than the cultivated variety characterized pod surface extracts of pod-borer-resistant cultivars. In addition, the ratio of the stilbene to quercetin-3-methyl ether was greater in the pod-borer-resistant cultivars. These findings are discussed in relation to the identification of chemical characters that can be used for crop improvement. PMID- 12775146 TI - Effect of iridoid glycoside content on oviposition host plant choice and parasitism in a specialist herbivore. AB - The Glanville fritillary butterfly Melitaea cinxia feeds upon two host plant species in Aland, Finland, Plantago lanceolata and Veronica spicata, both of which produce iridoid glycosides. Iridoids are known to deter feeding or decrease the growth rate of many generalist insect herbivores, but they often act as oviposition cues to specialist butterflies and are feeding stimulants to their larvae. In this study, two iridoid glycosides (aucubin and catalpol) were analyzed by micellar electrokinetic capillary chromatography. We measured the spatial and temporal variation of iridoid glycosides in natural populations of the host plants of M. cinxia. We also analyzed the aucubin and catalpol content in plants in relation to their use by ovipositing females, and in relation to the incidence of parasitism of M. cinxia larvae in natural populations. The mean concentrations of aucubin and catalpol were higher in P. lanceolata than in V. spicata, and catalpol concentrations were higher than aucubin concentrations in both host species. Plantago lanceolata individuals that were used for oviposition by M. cinxia had higher aucubin concentrations than random plants and neighboring plants. Additionally, oviposition and random plants had higher catalpol concentrations than neighboring plants, indicating that ovipositing females select for high iridoid glycoside plants or that oviposition induces iridoid glycoside production in P. lanceolata. Parasitism by the specialist parasitoid wasp Cotesia melitaearum occurred most frequently in larval groups that were feeding on plants with low concentrations of catalpol, irrespective of year, population, and host plant species. Therefore, parasitoids appear to avoid or perform poorly in host larvae with high catalpol content. PMID- 12775147 TI - Effects of Acacia condensed tannins on urinary parameters, body mass, and diet choice of an Acacia specialist rodent, Thallomys nigricauda. AB - The aim of this study was to investigate the dietary and physiological effects of condensed tannin ingestion on foregut fermenters, using Thallomys nigricauda, a folivorous rodent, as a model. We initially investigated the variability in physiological parameters, such as daily body mass (DMb), daily feed intake, daily fecal energy loss (FE), daily energy intake (DEI), daily urine pH, and daily urinary ammonia and urea concentrations, in response to different diets with low condensed tannin levels. This experiment was conducted to identify which physiological variables showed the least variation in the absence of tannin. In a second experiment, we investigated the response of the same dietary and physiological parameters to the effects of high dietary condensed tannin ingestion in T. nigricauda. We hypothesized that DMb, daily feed intake, FE, and DEI of T. nigricauda would be adversely affected by high dietary tannin content. We predicted that detoxification activity by T. nigricauda would increase at higher tannin levels. Ingestion of tannins affected the nutritional status of T. nigricauda, as shown by a decrease in body mass at high tannin levels. We also found that fewer ammonium ions were excreted in the urine by T. nigricauda, as would be expected if this were a means of regulating metabolic acidosis. The urine produced was more alkaline. This result indicates that T. nigricauda is not metabolizing these allelochemicals. Urea production was initially reduced, indicating conservation of bicarbonate ions that will neutralize blood acidity if there is detoxification. A diet choice experiment showed that tree rats avoid high tannin diets, even to the extent that they lose body mass on an alternative diet. This last-mentioned result is noteworthy because previous studies of the effects of tannins on herbivorous mammals have shown that there is physiological control rather than behavioral avoidance of the negative effects of tannin ingestion. PMID- 12775148 TI - Qualitative and quantitative variation in monoterpene co-occurrence and composition in the essential oil of Thymus vulgaris chemotypes. AB - Thymus vulgaris has a chemical polymorphism with six different chemotypes that show marked spatial segregation in nature. Although some populations have a single chemotype in majority, many have two or three chemotypes. In this study we analyze the quantitative variation among T. vulgaris populations in the percentage of oil composed of the dominant monoterpene(s) for each chemotype. In general, phenolic chemotypes (thymol and carvacrol), which occur at the end of the biosynthetic chain, have a significantly lower proportion of their oil composed of their dominant monoterpene than nonphenolic chemotypes (geraniol, alpha-terpineol, and linalool). This is due to the presence of high amounts of precursors (gamma-terpinene and paracymene) in the oil of phenolic chemotypes. The essential oil of the nonphenolic thuyanol chemotype has four characteristic monoterpenes that together make up a lower proportion of the oil than the single dominant monoterpene of the other nonphenolic chemotypes. For all chemotypes, the percentage composition of the dominant monoterpene decreased significantly at sites where the chemotype is not the majority type. This decrease is correlated with a significant increase in either the proportion of the two precursors for the thymol chemotype or the monoterpenes characteristic of the other chemotypes at the site. The latter result suggests that a plant with dominant genes is responsible for the production of different monoterpenes can produce several molecules. PMID- 12775149 TI - Novel detection of formylated phloroglucinol compounds (FPCs) in the wound wood of Eucalyptus globulus and E. nitens. AB - This study characterized the chemical responses of Eucalyptus globulus and Eucalyptus nitens to artificial inoculation with a basidiomycete decay fungus. Nine-year-old trees responded to mechanical wounding or inoculation with the decay fungus by producing new wound wood characterized by the presence of dark extractives 17 months after wounding. Analysis of crude wound wood extracts by HPLC coupled to negative ion electrospray mass spectrometry revealed the presence of a complex mixture of many unidentified formylated phlorglucinol compounds (FPCs), in addition to a diverse range of other polyphenolic compounds (hydrolyzable tannins, proanthocyanidins, flavanone glycoside, stilbene glycosides). Prior to this study, FPCs have only been reported from leaves and buds of Eucalyptus spp. Unequivocal evidence for the presence of macrocarpal A and B, and sideroxylonal A and B in the crude extracts was obtained, as well as evidence for a wide range of as yet unreported FPCs. Subsequent preliminary in vitro fungal and bacterial bioassays did not support an antimicrobial role for FPCs in host-pathogen interactions in eucalypts. PMID- 12775150 TI - Identification of odors from overripe mango that attract vinegar flies, Drosophila melanogaster. AB - Bioassays with a variety of overripe fruits, including mango, plum, pear, and grape, and their extracts showed that odors from overripe mango were most attractive to adult vinegar flies, Drosophila melanogaster. Combined gas chromatography-electroantennographic detection (GC-EAD) analyses of solid-phase microextraction (SPME) and Tenax extracts of overripe mango odors showed that several volatile compounds, including ethanol, acetic acid, amyl acetate, 2 phenylethanol, and phenylethyl acetate elicited significant EAG responses from antennae of female flies. Most of the volatile compounds in the extracts were identified by mass spectral and retention index comparisons with synthetic standards. In cage bioassays, lures with a blend of ethanol, acetic acid, and 2 phenylethanol in a ratio of 1:22:5 attracted six times more flies than any single EAG-active compound. This blend also attracted four times more flies than traps baited with overripe mango or unripe mango. However, in field trials, the blend was not as attractive as suggested by the laboratory bioassay. PMID- 12775151 TI - Oviposition response of Chironomus tepperi to nitrogenous compounds and bioextracts in two-choice laboratory tests. AB - Two-choice laboratory tests were used to investigate the oviposition response of Chironomus tepperi to a range of nitrogenous compounds and crude bioextracts. Responses to nitrogenous compounds varied in response to concentration. Ammonium nitrate did not influence oviposition at concentrations from 2 to 12 mg/liter. Hydroxylamine hydrochloride increased oviposition at 6 mg/liter, but had no effect at either 2 or 12 mg/liter. Sodium nitrate reduced oviposition at 2 mg/liter relative to the controls, but had no significant effect at 6 or 12 mg/liter. C. tepperi responded to many of the crude bioextracts, strongly avoiding oviposition in solutions containing homogenized chironomid larvae ( 1 final instar/100 ml; C. tepperi or Polypedilum nubiferum), and avoiding solutions conditioned by conspecific larvae at concentrations down to the equivalent of 1 final instar/100 ml over 24 hr. Homogenates of adult conspecifics had no effect on oviposition site selection. Homogenates of larval Culex annulirostris (Culicidae) deterred oviposition, but only at high concentrations (3 final instars/100 ml). Our results demonstrate that chemical cues from larval populations deter oviposition by C. tepperi females searching for newly flooded habitats where larval competition will be minimized. PMID- 12775152 TI - Sesamia calamistis calling behavior and its role in host finding of egg parasitoids Telenomus busseolae, telenomus isis, and Lathromeris ovicida. AB - In the Republic of Benin, the African pink stem borer Sesamia calamistis Hampson is kept under control by the two scelionid egg parasitoids Telenomus busseolae (Gahan) and T. isis (Polaszek). A third species, the trichogrammatid Lathromeris ovicida Risbec, although commonly obtained from egg batches, is of low importance. High egg and egg batch parasitism by Telenomus spp. at low host densities suggests excellent host-finding capacity. Previously it was hypothesized that pheromones produced by calling S. calamistis virgins play an important role in attracting the parasitoids to the area where oviposition is likely to occur. Thus, after determining the effect of age (1-4 days) on the calling rhythm of S. calamistis, 1-day-old virgin females were chosen to investigate the response of the parasitoids to calling and noncalling females and two empty controls, in a four-arm olfactometer. The results showed that calling started earliest and lasted longest with 1-day-old females, but was similar for 2 to 4-days-old females. The percentage of females calling was higher in early than latter scotophases. The highest percentage of active females was obtained between 8 and 9 hr after the beginning of the first scotophase. All three parasitoid species responded to calling females, while there were no differences between noncalling females and the empty controls. The response pattern was similar for the two Telenomus spp. but considerably weaker for L. ovicida, reflecting differing host specificity of the three parasitoid species. PMID- 12775153 TI - Volatile compounds released by disturbed and calm adults of the tarnished plant bug, Lygus lineolaris. AB - Volatile compounds released by disturbed and calm female and male Lygus lineolaris were collected and analyzed. Six major compounds were present in samples from disturbed bugs and from calm females: (E)-2-hexenal, 1-hexanol, (E) 2-hexenol, hexyl butyrate, (E)-2-hexenyl butyrate, and (E)-2,4-oxohexenal. (E)-2 hexenal was lacking in volatiles collected from calm males. Hexyl butyrate accounted for approximately 68% and 66% of volatiles released by agitated and calm females, and 87% and 88% of volatiles released by agitated and calm males, respectively. Blends released by disturbed insects differed quantitatively from blends released by calm insects, with amounts of compounds increasing 75-350 times in samples from disturbed insects. In static air bioassays, both females and males were repelled by natural volatiles collected from females and by five component [(E)-2,4-oxohexenal excluded] and six-component synthetic blends at doses of 1 and 10 bug-hours, indicating that these volatiles may serve an alarm or epideictic function, as well as a possible role as defensive allomones. Adults also avoided hexyl butyrate, (E)-2-hexenyl butyrate, (E)-2-hexenol, and (E)-2,4 oxohexenal, but not 1-hexanol and (E)-2-hexenal when compounds were assayed individually in static air bioassays at doses equal to 1 bug-hour. When tested over 1 day in two-choice cage trials, adults did not prefer untreated bean plants over bean plants surrounded by vials releasing up to 8.1 mg/hr (= 234 bug-hours) of the five-component synthetic blend. Therefore, the volatiles produced by disturbed adults would not be useful as a repellent for L. lineolaris. PMID- 12775154 TI - Phenotypic plasticity of Rhyzopertha dominica pheromone signaling: the effects of different hosts and presence of conspecific females on male produced aggregation pheromone. AB - Male Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae) release an aggregation pheromone consisting of a blend of two components, dominicalure 1 (D1) and Dominicalure 2 (D2). Pheromone from single insects, in different contexts, was collected and measured to determine if this signal is phenotypically plastic. Release rates were lowered when males were moved from maize grains to groundnut kernels or when moved from solitary occupation of maize grain to grain occupied by seven females. The pheromone release was increased again once these moves were reversed. The release of D1 was more affected than D2: thus, on groundnuts or in the presence of females, less pheromone was released and the proportion of D1 in the blend was lowered. Possible reasons for the modifications of the signal are discussed. PMID- 12775155 TI - Cuticular hydrocarbons of Drosophila birchii and D. serrata: identification and role in mate choice in D. serrata. AB - The cuticular hydrocarbon compositions of two sympatric species of Australian Drosophila in the montium subgroup of the melanogaster group that use cuticular hydrocarbons in mate recognition have been characterized. Drosophila birchii has 34 components in greater than trace amounts, with a carbon number range of C20 to C33. Drosophila serrata has 21 components above trace level and a carbon number range of C24 to C31. These two species share eight hydrocarbon components, with all but two of them being monoenes. For both species, the (Z)-9-monoenes are the predominant positional isomer. The hydrocarbons of D. birchii are n-alkanes, n alkenes (Z)-5-, (Z)-7-, (Z)-9-, and (Z)-11-), low to trace levels of homologous (Z,Z)-7,11- and (Z,Z)-9,13-dienes; and trace amounts of (Z,Z)-5,9-C25:2, a major component of D. serrata. Only one methyl branched hydrocarbon was detected (2 methyl C28), and it occurred at very low levels. The hydrocarbons of D. serrata are dominated by a homologous series of (Z,Z)-5,9-dienes, and notably, are characterized by the apparent absence of n-alkanes. Homologous series of (Z)-5-, (Z)-7-, and (Z)-9-alkenes are also present in D. serrata as well as 2-methyl alkanes. Drosophila serrata females display strong directional mate choice based on male cuticular hydrocarbons and prefer D. serrata males with higher relative abundances of the 2-methyl alkanes, but lower relative abundances of (Z,Z)-5.9 C24:2 and (Z)-9-C25:1. PMID- 12775156 TI - Sex pheromone of the vine mealybug, Planococcus ficus in Israel: occurrence of a second component in a mass-reared population. AB - Two pheromonal components were detected in airborne collections from the vine mealybug Planococcus ficus (Signoret) (Hemiptera: Pseudococcidae) mass-reared on potato sprouts. The compounds were identified as (S)-lavandulyl senecioate (I) and (S)-lavandulyl isovalerate (II) by GC and GC-MS by comparison with synthetic standards. Chiral GC analysis on a cyclodextrin column established their chirality. Compound I was identified recently as the sex pheromone of P. ficus in California. The attraction of vine mealybug males to both components I and II was demonstrated in a Petri dish bioassay and in a flight assay in the rearing chamber. Indoors, both compounds displayed a similar level of attractiveness to the mass-reared males. However, trials in a vineyard indicated that feral males were attracted only to compound I. Reanalysis of the airborne pheromone indicated that laboratory first generation daughters of females that were collected in the vineyard produce only (S)-lavandulyl senecioate (I). The relative amount of (S) lavandulyl isovalerate (II) increased gradually in each subsequent generation of P. ficus reared on potatoes. These findings indicate that feral P. ficus mealybugs produce and respond only to (S)-lavandulyl senecioate (I), whereas mealybugs that were reared in the laboratory on potato sprouts produce and respond to both (S)-lavandulyl senecioate (I) and (S)-lavandulyl isovalerate (II). PMID- 12775157 TI - Minor components in the sex pheromone of legume podborer: Maruca vitrata development of an attractive blend. AB - The legume podborer, Maruca vitrata (syn. M. testulalis) (F.) (Lepidoptera: Pyralidae) is a pantropical pest of legume crops. Sex pheromone was collected by gland extraction or trapping of volatiles from virgin female moths originating in India, West Africa, or Taiwan. Analysis by GC-EAG and GC-MS confirmed previously published findings that (E,E)-10,12-hexadecadienal is the most abundant EAG active component with 2-5% of (E,E)-10,12-hexadecadienol also present. At least one other EAG response was detected at retention times typical of monounsaturated hexadecenals or tetradecenyl acetates, but neither could be detected by GC-MS. Laboratory wind-tunnel bioassays and a field bioassay of blends of (E,E)-10,12 hexadecadienal with (E,E )-10,12-hexadecadienol and a range of monounsaturated hexadecenal and tetradecenyl acetate isomers indicated greatest attraction of males was to those including (E,E)-10,12-hexadecadienol and (E)-10-hexadecenal as minor components. In subsequent trapping experiments in cowpea fields in Benin, traps baited with a three-component blend of (E,E)-10,12-hexadecadienal and these two minor components in a 100:5:5 ratio caught significantly more males than traps baited with the major component alone, either two-component blend, or virgin female moths. Further blend optimization experiments did not produce a more attractive blend. No significant differences in catches were found between traps baited with polyethylene vials or rubber septa, or between lures containing 0.01 and 0.1 mg of synthetic pheromone. Significant numbers of female M. vitrata moths, up to 50% of total catches, were trapped with synthetic blends but not with virgin females. At present there is no clear explanation for this almost unprecedented finding, but the phenomenon may improve the predictive power of traps for population monitoring. PMID- 12775158 TI - Foraging mode and evolution of strike-induced chemosensory searching in lizards. AB - Strike-induced chemosensory searching (SICS) in lizards and snakes is a means of relocating prey by scent-trailing. The two main components of SICS are an elevated tongue-flick rate for vomerolfactory sampling after biting prey (PETF) and searching movements. In combination, these behaviors permit scent-trailing. Prey chemical discrimination, which is a prerequisite for SICS, is present in active foragers, but not in ambush foragers. Using comparative data. I show that searching movements and SICS have undergone correlated evolution with foraging mode and with prey chemical discrimination in lizards. This suggests that active foraging selects for prey chemical discrimination, which is then employed to search for escaped prey using the typical movements and tongue-flicking behaviors of active foragers. SICS in lizards is simply heightened active foraging after biting prey. In nonvenomous snakes, SICS is similar to that in lizards but is not restricted to active foragers. Only highly venomous snakes voluntarily release dangerous prey upon envenomation, pause to let the venom incapacitate the prey, and then relocate the prey by scent-trailing. PETF was observed in two ambush foragers and is not evolutionarily correlated with foraging mode or searching movements. Because it occurs in species lacking prey chemical discrimination, such PETF may be a response to gustatory cues or to internal chemicals not encountered on surfaces or trails of uninjured prey. PMID- 12775159 TI - Pheromonally mediated sexual isolation among denning populations of red-sided garter snakes, Thamnophis sirtalis parietalis. AB - Utilizing behavioral experiments and chemical analyses, we examined whether pheromonally mediated sexual isolation exists between denning populations of red sided garter snakes (Thamtnophis sirtalis parietalis) in Manitoba, Canada. Simultaneous choice tests conducted during the breeding season revealed that adult males from a hibernaculum in central Manitoba displayed a strong courtship preference for females from their own population over females from a hibernaculum in western Manitoba, whereas males from the western Manitoba hibernaculum showed no such preference. In addition. trailing experiments testing the response of males from the two hibernacula to familiar and unfamiliar female trails showed similar results, demonstrating that the observed male preference is mediated through chemical cues. Subsequent chemical analysis of the female sexual attractiveness pheromone. a homologous series of long-chain saturated and (omega 9 cis-unsaturated methyl ketones responsible for eliciting male courtship behavior and trailing behavior in garter snakes, showed significant variation in the composition of the pheromone between the two populations. Specifically, the two populations varied in the relative concentrations of individual unsaturated methyl ketones expressed by females. These results suggest that sexual isolation exists to a degree among denning populations of red-sided garter snakes due to variation in the expression of the female sexual attractiveness pheromone. PMID- 12775160 TI - (R)-acetoin-female sex pheromone of the summer chafer Amphimallon solstitiale (L.). AB - Extracts of Amphimallon solstitiale (L.), a well known, widely distributed and rather common European scarab beetle, were analyzed by GC-MS and GC-EAD. Acetoin (R):(S) > 9:1 - as well as 2,3-butanediol - (2R,3R): (2S,3S):meso = 1:1:9 - were present in extracts of both males and females. Although (2S,3S)-butanediol did not show any EAD activity, the other compounds elicited strong responses exclusively with male antennae. In contrast, several EAD active green leaf volatiles were detected equally well by male and female antennae. During preliminary field bioassays, (R)-acetoin was highly attractive to swarming males, whereas neither rac-acetoin nor the 2,3-butanediols showed activity. Therefore, (R)-acetoin is the female sex pheromone of A. solstitiale. PMID- 12775161 TI - Modern draw-over anesthetic vaporizers used to deliver anesthesia in austere and battlefield conditions. PMID- 12775162 TI - Possible systemic reaction to topical application of fluorescein dye. PMID- 12775163 TI - Explosive outbreaks of gastroenteritis in the shipboard environment attributed to Norovirus. PMID- 12775164 TI - MEDRET. PMID- 12775165 TI - The Greek armed forces are vulnerable to HAV infection. PMID- 12775166 TI - Drug addiction in America: challenges and opportunities. PMID- 12775167 TI - Operation Castle Cascade: managing multiple casualties from a simulated chemical weapons attack. AB - In the wake of the recent terrorist attack on the United States, there is an ever increasing need for the defense against weapons of mass destruction. The use of explosive devices in combination with chemical agents could result in a community disaster with multiple traumatic and medical injuries. Military medical personnel may be the first called upon due to their unique training and equipment. Operation Castle Cascade was a large-scale exercise on a military instillation involving the apprehension of hostages and detonation of an explosive device containing dimethyl sulfate. We will provide details on the medical management of 50 patients with simulated chemical and traumatic injuries. Issues relating to on site chemical identification, triage, decontamination, treatment, casualty collection, and transportation of casualties are addressed in this article. PMID- 12775168 TI - Infections of febrile neutropenic patients in malignant hematological diseases. AB - We observed 71 febrile, neutropenic episodes in 25 oncohematological patients after chemotherapy during a 3-years period from 1995 to 1997. Three patients died because of infections (pneumonia with septic shock, gram-negative bacteremia and sepsis, pseudomembranous colitis and diffuse peritonitis) at the period of prolonged, deep neutropenia (absolute neutrophil count < 100/mm3). During the 71 febrile, neutropenic episodes, we observed 24 bacteremia (33.8%) and 1 fungemia (1.4%). There were 35 cases of microbiologically documented and 12 cases of clinically documented infections. In 24 patients, the origin of fever was unknown. We analyzed the characteristics of infections, microbes and their susceptibility conditions, and the efficacy of empiric antimicrobial therapy. PMID- 12775169 TI - Aeromedical transfer to Belgium of severely burned patients during the initial days following the Volendam fire. AB - Nineteen critical burn patients were transferred to burn centers in Belgium soon after an indoor fire in The Netherlands. This evacuation was done by helicopter and by ambulance. The first arterial blood gases and pH measurements of the patients on arrival in the burn centers were analyzed and compared. All patients were in metabolic acidosis, but the respiratory component to the pH and the resulting pH was variable. Although an efficient fluid resuscitation to maintain an adequate urine output and arterial pressure is essential, the arterial blood gases and pH should also be monitored during the early evacuation of severe burn patients. Because hyperkaliemia is associated with a low pH value, we suggest that during the evacuation of these patients the ventilation setting should maintain the pH between 7.35 and 7.45. The comparison of the measurements between the group of patients transferred by air and the group transferred by road showed here differences in favor of a road transfer. PMID- 12775170 TI - Use of biomarkers of heavy drinking in health care practice. AB - Several biochemical tests are available to screen for heavy drinking in patients. Most of these (e.g., liver function tests and macrocytic volume) rise due to adverse effects of alcohol on organs or the organ's functions. Carbohydrate deficient transferrin, however, seems to elevate because of the direct action of alcohol or one of its metabolites. A second valuable contribution of biomarkers is identification of relapse events in recovering alcoholics that either would not be voluntarily reported or reported only much later. Finally, providing feedback to patients in treatment based on their biomarker values and changes in them during the course of treatment can enhance motivation, a key component in recovery from alcohol problems. The current article offers a clinical rationale for routine use of biomarkers of heavy drinking in health care practice and proposes recommendations on how they might be best used. PMID- 12775171 TI - U.S. military officer participation in the Centers for Disease Control and Prevention's Epidemic Intelligence Service (1951-2001). AB - The Epidemic Intelligence Service (EIS) was created in 1951 to provide epidemiologists to investigate natural and intentional disease epidemics. From an initial class of 23 U.S. citizens, the program has evolved into a globally recognized, hands-on learning experience, accepting approximately 65 to 75 new officers each year. The first U.S. military epidemic intelligence service officer (EISO) was accepted into the program in 1994. Since that time, 12 such officers have completed, or have begun, EIS training. They have comprised 2.1% of all EISOs from 1994 to 2001 and 0.47% of all EISOs. This total has included nine Air Force veterinarians, one Army veterinarian, one Army physician, and one Navy physician. Each military EISO had the opportunity to lead investigations of significant public health events (e.g., Ebola, monkeypox, malaria, Nipah virus, West Nile fever, and anthrax outbreaks). All graduates from the military returned to active duty assignments in operational medical units, research institutes, or the intelligence community. PMID- 12775172 TI - Use of the stages of change model in improving nutrition and exercise habits in enlisted Air Force men. AB - Thirty-nine U.S. Air Force enlisted men participated in a randomized controlled study that evaluated an experimental program designed to enhance fitness. Subjects were assigned to either a treatment (access to the program) or control (no access to program) group for 6 months. To improve treatment group fitness (as measured by VO2, subjects received individually tailored information (based on a behavior change model) via the Internet to encourage adoption of positive diet and exercise behaviors. Results showed no significant effect in improving fitness in treatment. However, significant effects were observed for secondary outcomes such as weight (controls, +1.0 kg vs. treatment, -2.2 kg, p < 0.05), body mass index (+0.3 kg/m2 vs. -0.7 kg/m2, p < 0.05), and percent body fat (+0.6% vs. 1.5%, p < 0.001). The data suggest that the individually tailored exercise information was not effective in encouraging sufficient exercise intensity to impact fitness. However, the dietary tailored information appears to have encouraged the adoption of more positive nutritional practices as demonstrated by the beneficial effects reflected in secondary outcomes. PMID- 12775173 TI - Breastfeeding experiences of active duty military women. AB - In the last 2 decades there has been an increase in both women in the workforce and women who breastfeed. All women in the workforce may experience a variety of issues when trying to combine breastfeeding and employment. Active duty women may experience issues similar to civilian working mothers, as well as issues specific to military service. The purpose of this qualitative study was to explore the breastfeeding experiences of active duty military women. An interactive interview process was used with a purposive sample obtained from the breastfeeding population of a Midwestern Air Force base. Overall, the nine women included in this study will provide health care professionals with knowledge to address more effectively the needs of the active duty breastfeeding mother. PMID- 12775174 TI - Premature labor and birth: influence of rank and perception of fatigue in active duty military women. AB - The purpose of this study was to determine whether stress, sleep disturbance, or fatigue at 22 to 26 weeks of gestation is related to the incidence of preterm labor and birth in military women. Data were collected prospectively by questionnaires measuring factors contributing to preterm labor and birth. Hours worked per week were ascertained by telephone until the participants delivered. Postpartum medical records were reviewed for pregnancy outcomes. There was a trend for preterm labor to be associated with lower perceived fatigue severity, low sleep disturbance, and more negative life events. Officer rank was related to both preterm labor and delivery. Findings indicate that the perception of fatigue may be protective against preterm birth and that military officer rank is a risk factor for preterm labor and birth. PMID- 12775175 TI - Effects of pediatric blood lead surveillance on Navy population health (1995 2001). AB - Navy medicine instituted a pediatric lead surveillance program in 1995 because of a public health concern over pediatric blood lead levels. This program included local sampling of pediatric populations and a central database at the Navy Environmental Health Center. That database contains 38,502 samples from 1995 through 2001 with 1.6% above 10 microg/dL. The number of samples submitted and percentage with elevated lead levels has steadily decreased during the 6-year reporting period. Rates above 10 microg/dL for children considered at high risk were similar for those residing on (2.4%) and off base (2.5%). Rates above 10 microg/dL for children considered at low risk, living off base, were 1.3%, whereas those living on base were 0.4% (relative risk = 3.44, p = 0.00, 95% confidence interval = 2.34-5.20). Several locations were identified as having higher risk for elevated blood levels that warrant continued surveillance. PMID- 12775176 TI - Predictors of successful physical readiness testing under the new standard: OPNAV Instruction 6110.1F. AB - A new physical readiness testing (PRT) instruction, OPNAV Instruction 6110.1F, specifying tougher standards compared with the previous standard was issued in May 2000. The purpose of this research was to describe differences in PRT results under the old and new standards, to describe body mass index (BMI) results, to compare BMI results with body composition assessment results, and to elucidate predictors of successful PRT results under the new standards. Using a retrospective cohort design, cross-sectional analysis was performed on data from 1,564 active duty subjects at the Naval Medical Center Portsmouth. Whereas, under the old standards, the distribution of PRT results was skewed toward high scores, the distribution of scores was normal under the new standards. BMI results demonstrated a problem with obesity in the cohort and correlated poorly with body composition assessment. Predictors of successful PRT were normal/underweight BMI, body composition within standards, officer rank, and age > or = 30 years. PMID- 12775177 TI - Socioenvironmental context of sexual trauma and well-being of women veterans. AB - The incidence of episodes of harassment and rape among military populations has only recently been examined. In the present study, a sample of 336 female veterans in a primary care setting was assessed. The incidences of lifetime sexual victimization, anxiety, depression, and impact of trauma for victims of specific trauma contexts are presented. Results of the study indicated that female veterans with a history of cumulative rape experiences and civilian rape experiences are more at risk for anxiety and depression than those with only a military experience of rape. No significant differences were found for impact of event scores for different contexts of rape, however. Reporting of trauma was not associated with psychological well-being for women veterans. The results highlight the role of the socioenvironmental context of abuse as an important variable to examine, especially in military populations. PMID- 12775178 TI - Varicella susceptibility and validity of history among U.S. Coast Guard recruits: an outbreak-based study. AB - During a varicella outbreak among U.S. Coast Guard recruits, we examined varicella susceptibility serologically and evaluated validity of disease history. Recruits completed a questionnaire to obtain information on demographics, history of varicella disease, and varicella vaccination. Serological testing for varicella-zoster virus immunoglobulin G antibodies was conducted using an enzyme linked immunosorbant assay. Among 513 recruits, 21 (4.1%) were seronegative to varicella-zoster virus. Recruits born in Puerto Rico were more likely than recruits born in the U.S. states to be susceptible (prevalence ratio, 4.3; 95% confidence interval, 1.4%, 13.1%). A positive disease history was highly predictive of positive serology (99.1%); however, 73% of those with a negative or uncertain history were also immune. Four (19%) susceptible recruits reported a positive varicella history. Although immunity among recruits was high, varicella outbreaks may occur in closed adult settings due to the high risks of exposure and transmission. Varicella vaccination can prevent these costly, disruptive outbreaks. PMID- 12775179 TI - The health effects of peacekeeping (Bosnia, 1992-1996): a cross-sectional study- comparison with nondeployed military personnel. AB - OBJECTIVE: Our goal was to test the hypothesis that United Kingdom soldiers who were deployed to Bosnia had worse health than a nondeployed control group. METHODS: We used data from a cross-sectional study designed to examine the health effects of service in the Persian Gulf War, which collected data in 1997 to 1998. We compared the two control groups--(personnel who were deployed to Bosnia and a nondeployed control group of military personnel (Era))--on a number of health related outcomes, including physical functioning, symptoms and ailments, psychological health, fatigue, and post-traumatic stress reactions. RESULTS: The response rate for the Bosnia cohort was 62.9% and for the Era group 61.9%. A proportion of the Bosnia group had served in the Persian Gulf War and was found to have considerably worse health outcomes than the remaining Bosnia group or the Era group. The Bosnia group who had not served in the Persian Gulf War had broadly similar health outcomes to the Era group. The main differences were that the Bosnia-only group consumed more alcohol and reported more fatigue, hay fever, weight gain, irritability, avoidance, and night sweats. Apart from heavy alcohol consumption, the magnitude of these differences was small. The Bosnia-only group had slightly better physical functioning than the Era group, and there were two other symptoms and one ailment which were less common in the Bosnia-only group than in the Era group. CONCLUSIONS: This study indicates that the health of United Kingdom military personnel who served in Bosnia from 1992 to 1996 was generally good in 1997 to 1998. However, further surveillance of veterans of the Balkan's War is required in the light of recent concerns. PMID- 12775180 TI - Mortality and postcombat disorders: U.K. veterans of the Boer War and World War I. AB - This study seeks to investigate the mortality rates of U.K. servicemen with postcombat syndromes following the Boer War and World War I. Random samples of veterans awarded war pensions for either disordered action of the heart (DAH) or neurasthenia/shellshock were compared with gunshot wounded ex-servicemen as controls. The destruction of pension records has led to reliance on groups of the longest lived veterans, which diminishes their representative qualities. Study groups were matched by rank and level of disability. With the exception of DAH cases in World War I, no statistically significant difference in mortality rates was found using Cox proportional hazards. The same DAH subjects were then compared with gunshot wound controls whose disability had been assessed 20% higher, and no statistically significant difference was seen. The reason why World War I veterans with DAH had a reduced life expectancy remains unclear, although it is possible that physician bias in assessment and the termination by the Ministry of Pensions of awards granted to healthy cases may have been factors. Postcombat disorders suffered by U.K. servicemen after the Boer War and World War I were not generally associated with an increased mortality. PMID- 12775181 TI - Penetrating liver war injury: a report on 172 cases. AB - During the 4-year military conflict in Croatia, we treated operatively 7,928 casualties. Of those casualties, 172 (2.2%) had penetrating liver injury, mostly sustained by explosive devices. Of these injuries, 90.7% were associated with the trauma of other abdominal and extra-abdominal organs. Seventy-five percent of injuries belonged to grades III and IV on the Liver Injury Scale. The main method of treatment was debridement with ligation of severed vessels and bile ducts. In 8.1% of cases with detrimental bleeding, we used liver packing. Fifty percent of these patients have survived but with a high incidence of septic complications. This method was proven salutary in the most detrimental injuries that could not be treated in any other way. Postoperative hemorrhage and intra-abdominal abscesses were complications that needed surgical and ultrasound-guided aspiration, respectively. Numerous heavy injuries of the liver combined with associated trauma of other vital organs are responsible for the high mortality rate of 28.5%. PMID- 12775182 TI - Occlusion of a wire-reinforced endotracheal tube in an almost completely edentulous patient. AB - Wire-reinforced endotracheal tubes have been advocated for use where endotracheal tube kinking is a risk. We report on a 79-year-old nearly edentulous male patient in a weakened state who managed to partially obstruct a wire-reinforced endotracheal tube, despite the presence of a soft bite block. The risk of kinking wire-reinforced endotracheal tubes is not mitigated simply because the patient is edentulous. Good monitoring, vigilance by providers and the use of a solid bite block remains critical in the care of these patients. PMID- 12775183 TI - A descriptive analysis of hypertension and affiliated therapies in a military retiree population (ages 40-85 years) at Camp Lejeune, North Carolina. AB - Because current methods of tracking hypertension are time-consuming and expensive, the prevalence of hypertension in retired military populations is often unknown. Tracking military retiree hypertension is important because it can help to conserve scarce resources and save lives by preventing strokes and other diseases. Intended as a case study, an alternative method of tracking hypertension using pharmacy records and ambulatory data system records to confirm diagnosis of hypertension was used to determine the prevalence of hypertension in a retired military population (ages 40-85 years). The results indicated that matching pharmacy data with ICD9 401 diagnosis data and medical records is a more efficient method of tracking hypertension. PMID- 12775184 TI - Fusarium proliferatum superficial suppurative thrombophlebitis. AB - Fungi are increasingly recognized as microorganisms causing superficial suppurative thrombophlebitis being managed by aggressive surgical therapy and antifungal agents. We report a case of Fusarium proliferatum superficial suppurative thrombophlebitis in an immunocompetent patient with no evidence of systemic infection and who responded to venotomy alone. PMID- 12775186 TI - Selective entorhinal and nonselective cortical-hippocampal region lesions, but not selective hippocampal lesions, disrupt learned irrelevance in rabbit eyeblink conditioning. AB - Prior experiments, as well as computational models, have implicated the hippocampal region in mediating the influence of nonreinforced stimulus preexposure on subsequent learning. Learned irrelevance (LIRR) is a preexposure task in which uncorrelated preexposures to the conditioned stimulus (CS) and the unconditioned stimulus (US) produce a retardation of subsequent CS-US conditioning. In the work presented here, we report the results of tests of LIRR in eyeblink conditioning in rabbits with sham lesions, nonselective cortical hippocampal region lesions, selective hippocampal lesions, and selective entorhinal lesions. Sham-lesioned rabbits that had been preexposed to the CS and the US exhibited slower acquisition of conditioned response, as compared with context-preexposed controls. Nonselective cotical-hippocampal region lesions disrupted LIRR, whereas selective hippocampal lesions had no detrimental effect on LIRR. Selective entorhinal lesions disrupted LIRR. These findings fit other recent empirical findings and theoretical predictions that some classical conditioning tasks previously thought to depend on the hippocampus depend, rather, on the entorhinal cortex. PMID- 12775187 TI - Distinguishing states of awareness from confidence during retrieval: evidence from amnesia. AB - Two experiments were conducted to determine whether recollective experience is distinguishable from confidence. In Experiment 1, we tested college participants in a within-subjects design and replicated Gardiner and Java's (1990) findings from a between-subjects design. We observed higher remember judgments for words than for nonwords, but higher know judgments for nonwords than for words. For confidence judgments, we obtained greater sure than unsure responses for both words and nonwords. In Experiment 2, we tested amnesic participants and matched control participants. Control participants produced the same pattern of results as college participants, but the results of amnesic participants diverged in an important way. For confidence judgments, the amnesic participants, like the control and college participants, made more sure than unsure judgments to both words and nonwords. But for recollective judgments, amnesic participants did not produce the crossover interaction for words and nonwords. This striking difference between the performance of memory-intact and amnesic participants demonstrates that recollective judgments and confidence that accompany retrieval are not isomorphic psychological experiences. PMID- 12775185 TI - The impact of synaptic depression following brain damage: a connectionist account of "access/refractory" and "degraded-store" semantic impairments. AB - Neuropsychological studies of patients with acquired semantic impairments have yielded two distinct and contrasting patterns of performance in a spoken word/picture-matching task (Warrington & Cipolotti, 1996). Patients labeled access/refractory are strongly influenced by presentation rate, semantic relatedness of distractors, and repetition, yet they seem relatively unaffected by lexical frequency. Degraded-store patients, on the other hand, are strongly affected by lexical frequency but are less affected by presentation rate, semantic relatedness, or repetition. Our account of these patterns of performance is based on the distinction between two different types of neurological damage: (1) damage to neuromodulatory systems that function to amplify neural signals while suppressing normal refractory-like effects and (2) damage to connections between groups of neurons that encode semantic information and are sensitive to frequency/familiarity. We present a connectionist model that learns to map spoken word input to semantic representations and that incorporates a particular form of neural refractoriness referred to as synaptic depression, as well as a simple form of neuromodulation. We show that the model is capable of accounting for the contrasting patterns of semantic impairment under these two different forms of damage and, furthermore, demonstrate how it is capable of handling several documented cases that are exceptions to the basic patterns of impairment. Several predictions and limitations of the present model are discussed. PMID- 12775188 TI - When objects lose their meaning: what happens to their use? AB - The 8 patients involved in this study were impaired on tests assessing knowledge of objects and on the demonstration of their use. The patients' success in object use was significantly correlated with their knowledge about the objects, providing further evidence that conceptual knowledge plays a key role in object use. Having a recipient present improved performance in the moderately impaired patients, suggesting that a certain level of conceptual knowledge must remain for the additional information to be beneficial. Although overall accuracy in using the target objects was not related to our measures of affordance, the specific aspects of use afforded by the objects' structures were relatively impervious to semantic impairment, suggesting a role for affordance information when object specific knowledge is disrupted. The patients' familiarity with the objects was an important predictor of performance. Finally, despite good performance on tests of mechanical problem solving, the patients showed very little evidence of employing these skills in their interactions with real objects. PMID- 12775189 TI - Event-related potentials of emotional memory: encoding pleasant, unpleasant, and neutral pictures. AB - Emotional events tend to be remembered better than nonemotional events. We investigated this phenomenon by measuring two event-related potential (ERP) effects: the emotion effect (more positive ERPs for pleasant or unpleasant stimuli than for neutral stimuli) and the subsequent memory effect (more positive ERPs for subsequently remembered items than for subsequently forgotten items). ERPs were measured while subjects rated the emotional content of pleasant, unpleasant, and neutral pictures. As was expected, subsequent recall was better for pleasant and unpleasant pictures than for neutral pictures. The emotion effect was sensitive to arousal in parietal electrodes and to both arousal and valence in frontocentral electrodes. The subsequent memory effect at centroparietal electrodes was greater for emotional pictures than for neutral pictures during an early epoch (400-600 msec). This result suggests that emotional information has privileged access to processing resources, possibly leading to better memory formation. PMID- 12775190 TI - Orbitofrontal cortex and dynamic filtering of emotional stimuli. AB - Event-related potentials (ER) were recorded in response to mildly aversive somatosensory and auditory stimuli. Patients with orbitofrontal lesions exhited enhanced ERPs (i.e., P3 amplitudes), as compared with control subjects. Moreover, these patients did not habituate to somatoensory stimuli across blocks of trials. The results were specific to orbitofrontal damage, since patients with damage to the dorsolateral prefontal cortex did not exhibit enhanced P3 amplitudes. These findings suggest damage to the orbitofrontal cortex impairs the ability to modulate or inhibit neural responses to aversive stimuli. The findings are couched in terms of dynamic filtering theory, which suggests that the orbitofrontal cortex is involved in the selection and active inhibition of neural circuits associated with emotional responses. PMID- 12775191 TI - Cortical activation during rhythmic hand movements performed under three types of control: an fMRI study. AB - Echoplanar fMRI was used to measure changes in cortical activation during the performance of a simple hand movement task under three types of voluntary control. Each of three imaging series alternated a task with rest: passive (in which the experimenter moved the hand), voluntary against low resistance, and voluntary against higher resistance. Contralateral activation was observed in the supplementary motor area (SMA), the primary motor cortex (M1), and the somatosensory cortex (S1) in all three tasks in each subject, whereas ipsilateral activation differed in each cortical region for each task. SMA had the widest prevalence of ipsilateral activation in all three tasks. In the M1, ipsilateral activation was observed in all but 1 subject in the two voluntary tasks but in only a few subjects in the S1 in any of the tasks. Quantitative changes in signal intensity and spatial extent of activation differentiated the voluntary tasks from the passive task and were most pronounced in the S1. PMID- 12775193 TI - Interventions for the prevention of brain atrophy in multiple sclerosis : current status. AB - The assessment of brain volume changes on serial magnetic resonance imaging (MRI) scans can provide an objective measure of the neurodegenerative component of multiple sclerosis (MS) pathology. Results from placebo-controlled and crossover clinical trials indicate that immunomodulating (e.g. recombinant interferon-beta [IFNbeta]-1a [Rebif] and IFNbeta-1b [Betaferon] and glatiramer acetate [Copaxone]) and immunosuppressive (e.g. cladribine and alemtuzumab) treatments for relapsing-remitting (RR) and secondary progressive MS lack substantial efficacy in preventing the development of brain atrophy, despite the marked effects of these treatments on clinical and MRI outcomes of disease activity. A modest but significant treatment effect on brain atrophy has been reported for patients with RRMS only in one trial of IFNbeta-1a (Avonex) and in another study of long-term corticosteroid therapy.Failure to find a significant treatment benefit in preventing brain atrophy might be the result of inadequate trial designs, including their relatively short durations, which may not be adequate to reveal beneficial effects in a chronic disease like MS. Alternatively, such a failure might indicate that treatments proven to be effective in reducing MS related inflammation are unable to act with the same efficacy on the most severe and disabling pathological substrates of the disease. The modest correlation between MRI enhancement frequency and brain atrophy observed in the placebo groups of several trials also fits with the concept that the suppression of inflammatory activity in MS is not fully and rapidly associated with a similar effect on the global neurodegenerative process of the disease. PMID- 12775192 TI - Mechanism of action of St John's wort in depression : what is known? AB - Extracts of Hypericum perforatum L. (St John's wort) are now successfully competing for status as a standard antidepressant therapy. Because of this, great effort has been devoted to identifying the active antidepressant compounds in the extract. From a phytochemical point of view, St John's wort is one of the best investigated medicinal plants. A series of bioactive compounds has been detected in the crude material, namely flavonol derivatives, biflavones, proanthocyanidines, xanthones, phloroglucinols and naphthodianthrones. Although St John's wort has been subjected to extensive scientific studies in the last decade, there are still many open questions about its pharmacology and mechanism of action. Initial biochemical studies reported that St John's wort is only a weak inhibitor of monoamine oxidase-A and -B activity but that it inhibits the synaptosomal uptake of serotonin, dopamine and noradrenaline (norepinephrine) with approximately equal affinity. However, other in vitro binding assays carried out using St John's wort extract demonstrated significant affinity for adenosine, GABA(A), GABA(B) and glutamate receptors. In vivo St John's wort extract leads to a downregulation of beta-adrenergic receptors and an upregulation of serotonin 5 HT(2) receptors in the rat frontal cortex and causes changes in neurotransmitter concentrations in brain areas that are implicated in depression. In studies using the rat forced swimming test, an animal model of depression, St John's wort extracts induced a significant reduction of immobility. In other experimental models of depression, including acute and chronic forms of escape deficit induced by stressors, St John's wort extract was shown to protect rats from the consequences of unavoidable stress. Recent neuroendocrine studies suggest that St John's wort is involved in the regulation of genes that control hypothalamic pituitary-adrenal axis function. With regard to the antidepressant effects of St John's wort extract, many of the pharmacological activities appear to be attributable to the naphthodianthrone hypericin, the phloroglucinol derivative hyperforin and several flavonoids. This review integrates new findings of possible mechanisms that may underlie the antidepressant action of St John's wort and its active constituents with a large body of existing literature. PMID- 12775195 TI - Sleep attacks and dopamine agonists for Parkinson's disease: what is currently known? AB - The aetiology of sleep disturbances in patients with Parkinson's disease is multifactorial. Medications, the disease process and underlying sleep disorders may contribute to sleepiness in patients with the disease. Somnolence, excessive daytime sleepiness and sleep attacks appear to be more common in patients with Parkinson's disease who are treated with dopamine receptor agonists than in those who are treated with other antiparkinsonian agents, although virtually all dopaminergic antiparkinsonian medications may contribute to sleepiness. Somnolence caused by dopamine agonists may be dose related and occurs most frequently during the dose-escalation phase. Somnolence may also emerge or worsen after a period of time on a stable dose. Patients with Parkinson's disease and caregivers should be informed about the risk of sleepiness and sleep attacks associated with dopaminergic medications and the potential implications for driving safety. PMID- 12775194 TI - Management of neurocysticercosis. AB - Neurocysticercosis is a common cause of neurological disease in developing countries and a major cause of epilepsy worldwide. A unique characteristic of human neurocysticercosis is that the living parasite is very well tolerated in human brain, so symptoms and clinical disease primarily result from death of the organism and accompanying inflammatory reaction in the human CNS. Among the diverse clinical manifestations of human neurocysticercosis, seizures are the most common, but other clinical problems occur, depending upon the localisation and viability of the parasite. Although both praziquantel and albendazole are effective agents, there is controversy about their role in several forms of the disease. Systematic reviews have pointed out the limited quality of available data on therapy. At a recent international conference convened to develop guidelines for treatment of this disease, areas of consensus and disagreement on the role of antiparasitic therapy were discussed. It was clear to all that cysticercosis cannot be regarded as a single disorder; treatment needs to be modified based on the location and number of cysticerci and the host response. There was a strong consensus that there is no role for antiparasitic drugs in patients with only calcified lesions. Studies suggest that patients with single enhancing lesions will do well regardless of antiparasitic therapy. Antiparasitic drugs are contraindicated in patients with cerebral oedema (cysticercal encephalitis). Most experts strongly recommend antiparasitic therapy in patients with multiple subarachnoid cysticerci or giant cysticerci. In patients with ventricular cysticerci, endoscopic removal is the preferred therapy. However, recent evidence suggests that placement of a ventricular shunt followed by antiparasitic therapy is an acceptable alternative. Standard treatment for localization-related epilepsy is effective for seizures caused by cysticercosis. In general, seizures are easily controlled in this illness. While many controversies regarding the treatment of patients with neurocysticercosis were not resolved at the international consensus conference, participants did conclude that controlled prospective studies are required to define optimal therapy for the infection and that treatment of infected individuals must be individualised. PMID- 12775197 TI - Graduation day for education programs. PMID- 12775198 TI - Effectiveness of supervised physical therapy in the early period after arthroscopic partial meniscectomy. AB - BACKGROUND AND PURPOSE: Controversy exists about the effectiveness of physical therapy after arthroscopic partial meniscectomy. This randomized controlled trial evaluated the effectiveness of supervised physical therapy with a home program versus a home program alone. SUBJECTS: Eighty-four patients (86% males; overall mean age=39 years, SD=9, range=21-58; female mean age=39 years, SD=9, range=24 58; male mean age=40, SD=9, range=21-58) who underwent an uncomplicated arthroscopic partial meniscectomy participated. METHODS: Subjects were randomly assigned to either a group who received 6 weeks of supervised physical therapy with a home program or a group who received only a home program. Blinded test sessions were conducted 5 and 50 days after surgery. Outcome measures were: (1) Hughston Clinic questionnaire, (2) Medical Outcomes Study 36-Item Short-Form Health Survey (SF-36) and EuroQol EQ-5D (EQ-5D) questionnaires, (3) number of days to return to work after surgery divided by the Factor Occupational Rating System score, (4) kinematic analysis of knee function during level walking and stair use, and (5) horizontal and vertical hops. RESULTS: No differences between groups were found for any of the outcomes measured. DISCUSSION AND CONCLUSION: The results indicate that the supervised physical therapy used in this study is not beneficial for patients in the early period after uncomplicated arthroscopic partial meniscectomy. PMID- 12775199 TI - Treatment of erectile dysfunction by perineal exercise, electromyographic biofeedback, and electrical stimulation. AB - BACKGROUND AND PURPOSE: Only a few investigators have described the involvement of the perineal muscles in the process of human erection. The aim of this research was to evaluate a re-education program for men with erection problems of different etiologies. SUBJECTS AND METHODS: Fifty-one patients with erectile dysfunction were treated with pelvic-floor exercises, biofeedback, and electrical stimulation. RESULTS: The results of the interventions can be summarized as follows: 24 patients (47%) regained a normal erection, 12 patients (24%) improved, and 6 patients (12%) did not make any progress. Nine patients (18%) did not complete the therapy. On the basis of several variables, a prediction equation was generated to determine the factors that would predict the effect of the interventions. The outcome was most favorable in men with venous-occlusive dysfunction. DISCUSSION AND CONCLUSION: Comparison of the results of the physical therapy protocol reported here with those obtained for other interventions reported in the literature shows that a pelvic-floor muscle program may be a noninvasive alternative for the treatment of patients with erectile dysfunction caused by venous occlusion. PMID- 12775200 TI - Relationship of knee extension force to independence in sit-to-stand performance in patients receiving acute rehabilitation. AB - BACKGROUND AND PURPOSE: The ability to rise from a chair is important for independence in everyday life. This study was conducted to determine to what extent knee extension force (KEF) could explain independence in sit-to-stand (STS) performance from a standard chair. SUBJECTS AND METHODS: This was a descriptive and correlational study of patients receiving acute rehabilitation (N=107). Measurements of KEF of both lower extremities were obtained using manual muscle testing (MMT) and hand-held dynamometers (HHDs). The HHD measurements were normalized based on body weight and age- and sex-specific reference values. Measurements of the ability to stand without using the upper extremities (STS [without hands]) and to stand using the upper extremities (STS [with hands]) were correlated with the force measurements. RESULTS: The correlations (r) of the KEF measurements with STS success ranged from.652 to.708 for STS (without hands) and from.545 to.638 for STS (with hands). Body weight added to the explanation of STS (without hands) (R=.662) and STS (with hands) (R=.584) provided by KEF measured by HHD. The receiver operating characteristic curves showed that HHD (weight normalized) was the most sensitive and specific measure for explaining independence in STS. DISCUSSION AND CONCLUSION: Manual and dynamometric measurements of KEF are related to independence in STS. Measurements of KEF quantified with a dynamometer and normalized against body weight provided the most valid, specific, and sensitive cutoff point for explaining STS independence. PMID- 12775201 TI - Medial pontine hemorrhagic stroke. AB - BACKGROUND AND PURPOSE: This case report documents a rare opportunity to observe the motor function of an individual for nearly 6 months following a primary pontine hemorrhage in the medial pontine tegmentum of the brain stem. The purpose of this report is to illustrate how knowledge of the location of the hemorrhage, in conjunction with knowledge of brain-stem structure-function relationships, informs physical therapist examination and intervention. CASE DESCRIPTION: RM, a right-handed 81-year-old man with hypertension, had a hemorrhagic brain-stem stroke that severely compromised control of posture and whole-limb movements. Some residual ability to use the right hand and fingers remained, provided the trunk and right upper arm were stabilized. RM had undiminished intellectual abilities and unaltered memory because of sparing of cerebral cortices. RM's cognitive abilities, however, were obscured by severe impairments in interpersonal communication because of extensive damage to cranial nerve structures. Computed tomographic scans verified that the hematoma crossed the midline and was confined to the medial pontine tegmentum. DISCUSSION: We interpret motor deficits resulting from stoke in the medial pontine tegmentum in terms of damage to brain-stem descending motor systems and ascending somatosensory systems. Recognition of cognitive and residual motor abilities following brain-stem stroke can aid in the development of rehabilitation strategies. PMID- 12775202 TI - Acute care physical therapist evaluation and intervention for an adult after right hemispherectomy. AB - BACKGROUND AND PURPOSE: Hemispherectomy is performed to help control intractable seizures, yet little research quantifies and projects the clinical course of the impairments, functional limitations, and disabilities of patients who have undergone the procedure. This case report describes the physical therapist preoperative and postoperative examination, evaluation, and intervention during the acute hospitalization of an adult who underwent a right hemispherectomy. CASE DESCRIPTION: The patient was a 27-year-old man who had intractable seizures despite having tried multiple drug regimens and undergoing several surgical interventions after a brain injury at age 5 years. He underwent a right functional hemispherectomy and then had 9 postoperative physical therapy sessions during his acute hospitalization. OUTCOMES: The patient made rapid gains, surpassing all initial goals. At discharge, the patient had distal left-sided sensorimotor impairments, but he was able to ambulate 121.9 m (400 ft) with assistance to maintain his balance. DISCUSSION: The patient's posthemispherectomy recovery was rapid. His brain injury at a young age may have triggered preoperative transfer of function to the unaffected left hemisphere. The prognosis for this patient's improvements of impairments and functional limitations was better than initially expected, perhaps because of the redundant features and plasticity of the central nervous system. PMID- 12775203 TI - Economic evaluation: a useful research method. PMID- 12775204 TI - Diagnosing painful sacroiliac joints: A validity study of a McKenzie evaluation and sacroiliac provocation tests. AB - Research suggests that clinical examination of the lumbar spine and pelvis is unable to predict the results of diagnostic injections used as reference standards. The purpose of this study was to assess the diagnostic accuracy of a clinical examination in identifying symptomatic and asymptomatic sacroiliac joints using double diagnostic injections as the reference standard. In a blinded concurrent criterion-related validity design study, 48 patients with chronic lumbopelvic pain referred for diagnostic spinal injection procedures were examined using a specific clinical examination and received diagnostic intraarticular sacroiliac joint injections. The centralisation and peripheralisation phenomena were used to identify possible discogenic pain and the results from provocation sacroiliac joint tests were used as part of the clinical reasoning process. Eleven patients had sacroiliac joint pain confirmed by double diagnostic injection. Ten of the 11 sacroiliac joint patients met clinical examination criteria for having sacroiliac joint pain. In the primary subset analysis of 34 patients, sensitivity, specificity and positive likelihood ratio (95% confidence intervals) of the clinical evaluation were 91% (62 to 98), 83% (68 to 96) and 6.97(2.70 to 20.27) respectively. The diagnostic accuracy of the clinical examination and clinical reasoning process was superior to the sacroiliac joint pain provocation tests alone. A specific clinical examination and reasoning process can differentiate between symptomatic and asymptomatic sacroiliac joints PMID- 12775205 TI - The effect of physiotherapy treatment on oxygen consumption and haemodynamics in patients who are critically ill. AB - The purpose of this study was to determine metabolic and haemodynamic changes with and without physiotherapy treatment in haemodynamically stable, intubated and ventilated patients. This was a prospective, randomised cross-over study. Ten intubated, ventilated and haemodynamically stable patients underwent a 20 min physiotherapy treatment and a 20 min period of undisturbed side lying. Mean oxygen consumption (VO2mean) was measured on a minute-to-minute basis by indirect calorimetry. Mean arterial pressure (MAP) was recorded minutely from the indwelling arterial line and cardiac index (CI) was calculated from the indwelling pulmonary artery catheter. Time to recovery to within 5% of resting VO2 was also recorded. The results showed no significant increase in VO2mean with either positioning the patient in side lying or physiotherapy treatment (p = 0.17). Time to recovery to within 5% of baseline VO2 occurred within seven minutes for all patients and there was no significant difference between either physiotherapy treatment or positioning in side lying (p = 0.63). There were no significant differences in CI (p = 0.44) or MAP (p = 0.95) during physiotherapy treatment compared with undisturbed side lying. It is concluded that physiotherapy treatment does not significantly alter VO2mean or MAP and CI in stable intubated and ventilated patients. PMID- 12775206 TI - A systematic review of low level laser therapy with location-specific doses for pain from chronic joint disorders. AB - We investigated if low level laser therapy (LLLT) of the joint capsule can reduce pain in chronic joint disorders. A literature search identified 88 randomised controlled trials, of which 20 trials included patients with chronic joint disorders. Six trials were excluded for not irradiating the joint capsule. Three trials used doses lower than a dose range nominated a priori for reducing inflammation in the joint capsule. These trials found no significant difference between active and placebo treatments. The remaining 11 trials including 565 patients were of acceptable methodological quality with an average PEDro score of 6.9 (range 5-9). In these trials, LLLT within the suggested dose range was administered to the knee, temporomandibular or zygapophyseal joints. The results showed a mean weighted difference in change of pain on VAS of 29.8 mm (95% CI, 18.9 to 40.7) in favour of the active LLLT groups. Global health status improved for more patients in the active LLLT groups ( relative risk of 0.52; 95% CI 0.36 to 0.76). Low level laser therapy with the suggested dose range significantly reduces pain and improves health status in chronic joint disorders, but the heterogeneity in patient samples, treatment procedures and trial design calls for cautious interpretation of the results. PMID- 12775207 TI - Injury in the Australian sport of calisthenics: a prospective study. AB - The aims of this study were to determine the rate, anatomical regions, onset, severity, and type of injury in the sport of calisthenics and compare injuries reported by elite and non-elite participants. Prospective reports of injuries were collected over a 12-month period from 550 elite and non-elite calisthenics participants. The participants recorded the number of training sessions, competition, and performances per week, hours of training, and information on any injuries sustained each week during the survey period. Five hundred and fifty participants reported 190 injuries during the survey period, 0.4 injuries per participant year or 0.3 injured participants per participant year. The odds ratio of injury in the elite to the non-elite group was 2.0 (95% CI 1.3 to 2.9). Injuries to the lower back (32.4% of all injuries), hip thigh and groin (25.4% of all injuries) were most common. Activities involving lumbar extension (29.8% of all injuries and 61.0% of lower back injuries) were perceived by participants to have led to injury. In general, injuries were minor and mainly involved soft tissue structures (95.6% of all injuries). Participants had difficulty in identifying why their injuries had occurred. Calisthenics participants did not report high injury rates, but activities that involve lumbar extension are implicated in low back injuries and warrant further attention PMID- 12775208 TI - Arm positioning alters lung volumes in subjects with COPD and healthy subjects. AB - Subjects with chronic obstructive pulmonary disease (COPD) have difficulty performing arm exercise, particularly if the arms are unsupported and elevated. The purpose of this study was to evaluate the effect of arm position on static lung volumes in COPD and healthy subjects. Lung volumes were measured by plethysmography in nine COPD subjects (mean age +/- SD = 67.3 +/- 10.3 years; % pred FEV1 +/- SD = 39.7 +/- 10.9%) and nine healthy subjects (mean age +/- SD = 55.8 +/- 8.8 years; % pred FEV1 +/- SD = 102.9 +/- 12.2%) with the arms below 90 degrees shoulder flexion, at 90 degrees shoulder flexion and above 90 degrees shoulder flexion. In all subjects a significant increase in functional residual capacity (FRC) and reduction in inspiratory capacity (IC) was shown with arms above 90 degrees shoulder flexion when compared with both arms below 90 degrees shoulder flexion (mean increase in FRC (95% CI) was 0.17 L (0.06 to 0.27) for COPD and 0.29 L (0.11 to 0.47) for healthy subjects; mean reduction in IC (95% CI) was 0.24 L (0.1 to 0.38) for COPD and 0.45 L (0.22 to 0.68) for healthy subjects) and arms at 90 degrees shoulder flexion (mean increase in FRC (95% CI) was 0.15 L (0.01 to 0.29) for COPD and 0.22 L (0.11 to 0.34) for healthy subjects; mean reduction in IC (95% CI) was 0.14 L (0.01 to 0.26) for COPD and was 0.29 L (0.17 to 0.42) for healthy subjects). These changes may alter lung mechanics and, in COPD subjects, may affect their ability to perform arm exercise above shoulder height PMID- 12775209 TI - Debate on effectiveness of spinal manipulation may have opened a Pandora's box. (Comment on Ferreira M et al, Australian Journal of Physiotherapy 48: 277-284, Edmondston S, Australian Journal of Physiotherapy 49: 63-64 and Ferreira M et al, Australian Journal of Physiotherapy 49: 64.). PMID- 12775210 TI - Need to differentiate traditional Chinese acupuncture from other forms of acupuncture. (Comment on critically appraised paper, Australian Journal of Physiotherapy 49: 74.). PMID- 12775212 TI - Definition of structural elements in Plasmodium vivax and P. knowlesi Duffy binding domains necessary for erythrocyte invasion. AB - Plasmodium vivax and P. knowlesi use the Duffy antigen as a receptor to invade human erythrocytes. Duffy-binding ligands belong to a family of erythrocyte binding proteins that bind erythrocyte receptors to mediate invasion. Receptor binding domains in erythrocyte-binding proteins lie in conserved cysteine-rich regions called Duffy-binding-like domains. In the present study, we report an analysis of the overall three-dimensional architecture of P. vivax and P. knowlesi Duffy-binding domains based on mild proteolysis and supportive functional assays. Our proteolysis experiments indicate that these domains are built of two distinct subdomains. The N-terminal region from Cys-1-4 (C1-C4) forms a stable non-functional subdomain. The region spanning C5-C12 forms another subdomain, which is capable of binding Duffy antigen. These subdomains are joined by a protease-sensitive linker. Results from deletion constructs, designed for expression of truncated proteins on COS cell surface, show that regions containing C5-C8 of the Duffy-binding domains are sufficient for the binding receptor. Therefore the central region of Duffy-binding domains, which is flanked by two non-functional regions, is responsible for receptor recognition. Moreover, the minimal Duffy-binding region identified here is capable of folding into a functionally competent module. These studies pave the way for understanding the architecture of Duffy-binding domains and their interactions with host receptors. PMID- 12775214 TI - Overexpression of UDP-glucose dehydrogenase in Escherichia coli results in decreased biosynthesis of K5 polysaccharide. AB - The Escherichia coli K5 capsular polysaccharide (glycosaminoglycan) chains are composed of the repeated disaccharide structure: -GlcAbeta1,4-GlcNAcalpha1,4 (where GlcA is glucuronic acid and GlcNAc is N-acetyl-D-glucosamine). The GlcA, present in most glycosaminoglycans, is donated from UDP-GlcA, which, in turn, is generated from UDP-glucose by the enzyme UDP-glucose dehydrogenase (UDPGDH). The formation of UDP-GlcA is critical for the biosynthesis of glycosaminoglycans. To investigate the role of UDPGDH in glycosaminoglycan biosynthesis, we used K5 polysaccharide biosynthesis as a model. E. coli was transformed with the complete gene cluster for K5 polysaccharide production. Additional transformation with an extra copy of UDPGDH resulted in an approx. 15-fold increase in the in vitro UDPGDH enzyme activity compared with the strain lacking extra UDPGDH. UDP-GlcA levels were increased 3-fold in overexpressing strains. However, metabolic labelling with [14C]glucose showed, unexpectedly, that overexpression of UDPGDH lead to decreased formation of K5 polysaccharide. No significant difference in the K5 polysaccharide chain length was observed between control and overexpressing strains, indicating that the decrease in K5-polysaccharide production most probably was due to synthesis of fewer chains. Our results suggest that K5-polysaccharide biosynthesis is strictly regulated such that increasing the amount of available UDP-GlcA results in diminished K5 polysaccharide production. PMID- 12775213 TI - Protein-protein, protein-RNA and protein-lipid interactions of signal-recognition particle components in the hyperthermoacidophilic archaeon Acidianus ambivalens. AB - The signal-recognition particle (SRP) of one of the most acidophilic and hyperthermophilic archaeal cells, Acidianus ambivalens, and its putative receptor component, FtsY (prokaryotic SRP receptor), were investigated in detail. A. ambivalens Ffh (fifty-four-homologous protein) was shown to be a soluble protein with strong affinity to membranes. In its membrane-residing form, Ffh was extracted from plasma membranes with chaotropic agents like urea, but not with agents diminishing electrostatic interactions. Using unilamellar tetraether phospholipid vesicles, both Ffh and FtsY associate independently from each other in the absence of other factors, suggesting an equilibrium of soluble and membrane-bound protein forms under in vivo conditions. The Ffh protein precipitated from cytosolic cell supernatants with anti-Ffh antibodies, together with an 7 S-alike SRP-RNA, suggesting a stable core ribonucleoprotein composed of both components under native conditions. The SRP RNA of A. ambivalens depicted a size of about 309 nucleotides like the SRP RNA of the related organism Sulfolobus acidocaldarius. A stable heterodimeric complex composed of Ffh and FtsY was absent in cytosolic supernatants, indicating a transiently formed complex during archaeal SRP targeting. The FtsY protein precipitated in cytosolic supernatants with anti-FtsY antisera as a homomeric protein lacking accessory protein components. However, under in vitro conditions, recombinantly generated Ffh and FtsY associate in a nucleotide-independent manner, supporting a structural receptor model with two interacting apoproteins. PMID- 12775215 TI - Subsite structure of the endo-type chitin deacetylase from a deuteromycete, Colletotrichum lindemuthianum: an investigation using steady-state kinetic analysis and MS. AB - The endo-type chitin deacetylase (EC 3.5.1.41) from a deuteromycete, Colletotrichum lindemuthianum (ATCC 56676), catalyses the hydrolysis of the acetamido group of GlcNAc (2-acetamido-2-deoxy-D-glucose) residues in chitin or chito-oligosaccharides with a degree of polymerization (n) equal to or greater than 2. The steady-state kinetic parameters for the initial deacetylation reactions of (GlcNAc)(2-6) were determined using a direct, continuous spectrophotometric assay in combination with ESI-MS (electrospray ionization MS) analysis of the products. The dependence of the observed K(m) and k(cat)/K(m) on n suggests the presence of four enzyme subsites (-2, -1, 0 and +1) that interact with GlcNAc residues from the non-reducing end to the reducing end of the substrate. The turnover number (k (cat), 7 s(-1)) is independent of n and represents the intrinsic rate constant (k(int)) for the hydrolysis of the acetamido group in subsite 0. The subsite affinities for the GlcNAc residues were calculated from the observed k(cat)/K(m) values (A (-2), -11.0; A (-1), -1.5; A (0), -7.7; A (+1), -12.5 kJ x mol(-1)). The increments in the subsite affinities due to the recognition of the acetamido groups were calculated [DeltaDelta G ((N acetyl))=3.3, 0, 4.0 and 0 kJ x mol(-1) for subsites -2, -1, 0 and +1 respectively]. The steady-state kinetic parameters for the second deacetylation reaction of (GlcNAc)(4) were also determined using (GlcNAcGlcNAcGlcNGlcNAc) as the substrate. The comparison of the experimental and theoretical values (calculated using the subsite affinities) suggests that the mono-deacetylated substrate binds strongly in a non-productive mode occupying all four subsites, thereby inhibiting the second deacetylation reaction. PMID- 12775216 TI - Oxidized low-density lipoprotein induces calpain-dependent cell death and ubiquitination of caspase 3 in HMEC-1 endothelial cells. AB - Oxidized low-density lipoprotein (oxLDL) is known to induce apoptosis in endothelial cells, and this is believed to contribute to the progression of atherosclerosis. In the present study we made the novel observation that oxLDL induced death of HMEC-1 cells is accompanied by activation of calpain. The mu calpain inhibitor PD 151746 decreased oxLDL-induced cytotoxicity, whereas the general caspase inhibitor BAF (t-butoxycarbonyl-Asp-methoxyfluoromethylketone) had no effect. Also, oxLDL provoked calpain-dependent proteolysis of cytoskeletal alpha-fodrin in the HMEC-1 cells. Our observation of an autoproteolytic cleavage of the 80 kDa subunit of mu-calpain provided further evidence for an oxLDL induced stimulation of calpain activity. The Bcl-2 protein Bid was also cleaved during oxLDL-elicited cell death, and this was prevented by calpain inhibitors, but not by inhibitors of cathepsin B and caspases. Treating the HMEC-1 cells with oxLDL did not result in detectable activation of procaspase 3 or cleavage of PARP [poly(ADP-ribose) polymerase], but it did cause polyubiquitination of caspase 3, indicating inactivation and possible degradation of this protease. Despite the lack of caspase 3 activation, oxLDL treatment led to the formation of nucleosomal DNA fragments characteristic of apoptosis. These novel results show that oxLDL initiates a calpain-mediated death-signalling pathway in endothelial cells. PMID- 12775217 TI - Environmental stresses can alleviate the average deleterious effect of mutations. AB - BACKGROUND: Fundamental questions in evolutionary genetics, including the possible advantage of sexual reproduction, depend critically on the effects of deleterious mutations on fitness. Limited existing experimental evidence suggests that, on average, such effects tend to be aggravated under environmental stresses, consistent with the perception that stress diminishes the organism's ability to tolerate deleterious mutations. Here, we ask whether there are also stresses with the opposite influence, under which the organism becomes more tolerant to mutations. RESULTS: We developed a technique, based on bioluminescence, which allows accurate automated measurements of bacterial growth rates at very low cell densities. Using this system, we measured growth rates of Escherichia coli mutants under a diverse set of environmental stresses. In contrast to the perception that stress always reduces the organism's ability to tolerate mutations, our measurements identified stresses that do the opposite - that is, despite decreasing wild-type growth, they alleviate, on average, the effect of deleterious mutations. CONCLUSIONS: Our results show a qualitative difference between various environmental stresses ranging from alleviation to aggravation of the average effect of mutations. We further show how the existence of stresses that are biased towards alleviation of the effects of mutations may imply the existence of average epistatic interactions between mutations. The results thus offer a connection between the two main factors controlling the effects of deleterious mutations: environmental conditions and epistatic interactions. PMID- 12775218 TI - Influence of social support on cognitive function in the elderly. AB - BACKGROUND: Social support is important in daily activities of the elderly. This study tests the hypothesis that there is an association between social support and cognitive function among the elderly in a community setting. METHODS: Face-to face interviews were conducted in a cross-sectional stratified random sample of 4,993 elderly (> or =65 years) city residents. Using multiple regression analysis, we investigated the influence of social support on cognitive function. RESULTS: 12% were over 80 years old. 53.28% were men. 67.14% were married. Higher Short Portable Mental Status Questionnaire (SPMSQ) scores (higher score means better cognitive function) were associated with strong social support, as measured by marital status and perceived positive support from friends. Lower cognitive function was associated with older and with female respondents. Only instrumental activities of daily living (IADL) were statistically and negatively related to SPMSQ. Lower functional status was associated with lower cognitive function. Elders with grade school educations had lower SPMSQ scores than did elders with high school educations. CONCLUSIONS: In Taiwan, higher cognitive function in community-living elderly was associated with increased social support. Life-style management should provide social activities for the elderly to promote a better quality of life. PMID- 12775219 TI - An evaluation of the integration of non-traditional learning tools into a community based breast and cervical cancer education program: the Witness Project of Buffalo. AB - BACKGROUND: Breast and cervical cancer continue to represent major health challenges for African American women. among Caucasian women. The underlying reasons for this disparity are multifactorial and include lack of education and awareness of screening and early detection. Traditional educational methods have enjoyed varied success in the African American community and spawned development of novel educational approaches. Community based education programs employing a variety of educational models have been introduced. Successful programs must train and provide lay community members with the tools necessary to deliver strong educational programs. METHODS: The Witness Project is a theory-based, breast and cervical cancer educational program, delivered by African American women, that stresses the importance of early detection and screening to improve survival and teaches women how to perform breast self examination. Implementing this program in the Buffalo Witness Project of Buffalo required several modifications in the curriculum, integration of non-traditional learning tools and focused training in clinical study participation. The educational approaches utilized included repetition, modeling, building comprehension, reinforcement, hands on learning, a social story on breast health for African American women, and role play conversations about breast and cervical health and support. RESULTS: Incorporating non-traditional educational approaches into the Witness Project training resulted in a 79% improvement in the number of women who mastered the didactic information. A seventy-two percent study participation rate was achieved by educating the community organizations that hosted Witness Project programs about the informed consent process and study participation. CONCLUSION: Incorporating non-traditional educational approaches into community outreach programs increases training success as well as community participation. PMID- 12775220 TI - Information extraction from full text scientific articles: where are the keywords? AB - BACKGROUND: To date, many of the methods for information extraction of biological information from scientific articles are restricted to the abstract of the article. However, full text articles in electronic version, which offer larger sources of data, are currently available. Several questions arise as to whether the effort of scanning full text articles is worthy, or whether the information that can be extracted from the different sections of an article can be relevant. RESULTS: In this work we addressed those questions showing that the keyword content of the different sections of a standard scientific article (abstract, introduction, methods, results, and discussion) is very heterogeneous. CONCLUSIONS: Although the abstract contains the best ratio of keywords per total of words, other sections of the article may be a better source of biologically relevant data. PMID- 12775221 TI - Myoepithelial carcinoma of the salivary glands: behavior and management. AB - OBJECTIVE: To investigate the biological behavior and proper management of myoepithelial carcinomas of salivary glands. METHODS: Twenty-seven cases of myoepithelial carcinoma of salivary glands were retrospectively studied and their detailed clinical and follow-up data were presented. RESULTS: The subjects consisted of 17 men and 10 women aged 16 to 73 years (mean age: 51 years). The parotid gland was the most common site (n = 14) of cancer. Clinical features included extensive local growth, invasion of the surrounding tissues, infrequent cervical lymph node metastasis but high rates of distant metastasis, frequent/multiple recurrences and poor prognosis. CONCLUSIONS: Myoepithelial carcinomas of the salivary gland should be classified as high-grade malignancies. Early and radical surgery with close follow-up are essential for achieving favorable outcomes. Radiotherapy appears to be non-sensitive and elective neck dissection is generally unnecessary. PMID- 12775222 TI - Cloning the full-length cDNA of a gene responsible for vascular smooth muscle cell proliferation in atherogenesis and study of function. AB - OBJECTIVE: To clone the full-length cDNA of a gene responsible for vascular smooth muscle cell (v-SMC) proliferation in atherogenesis, and study its function. METHODS: Oxidized low density lipoprotein (ox-LDL) at optimal concentration was used as the stimulant to induce v-SMC proliferation in culture medium. A cDNA subtractive library of v-SMC proliferation specific to ox-LDL stimulation was established using subtractive hybridization technique. Methods, including blotting, Northern hybridization and gene sequencing, were used to clone new gene fragments. By using full-length cDNA screening and protein expression techniques, one full-length cDNA was cloned and its function was studied. RESULTS: One full-length cDNA was cloned. The new gene (Genbank AF 174647) expressed a 44 kDa protein, which might be associated with the activity of ox-LDL. CONCLUSION: The new gene cloned may be associated with SMC proliferation in atherogenesis. PMID- 12775223 TI - Management of enterocutaneous fistulas: 30-year clinical experience. AB - OBJECTIVES: To explore successful models of management of enterocutaneous fistulas and unresolved problems requiring further study. METHOD: Analysis of therapeutic results of 1168 cases treated in one center from January 1971 to December 2000. RESULTS: In this group of patients, the recovery rate was 93% and 37% of fistulas healed spontaneously after non-operative treatment. The mortality rate was 5.5%, most of which occurred due to sepsis. Of 659 cases receiving definitive operations for enteric fistula, 98% recovered. Recovery, mortality and operational success rates (94.2%, 4.4%, 99.7%) of cases treated between January 1985 and December 2000 were significantly better than those (90.4%, 8.2%, 95.5%) of cases treated earlier (January 1971-December 1984) (P < 0.05). CONCLUSIONS: The results from this study were better overall than those reported in previous literatures. The change in therapeutic strategy, improved technique in control of sepsis, rational nutritional support and careful monitoring of vital organs are the key reasons for improvement of managing enteric fistulas. However, increasing spontaneous closure of fistula, improving the therapeutic rate of specific enteric fistula (IBD or radiation enteritis) and performing definitive operations for enteric fistula at early stages are still problematic and require further study. PMID- 12775224 TI - Mapping cortical areas associated with Chinese word processing using functional magnetic resonance imaging. AB - OBJECTIVES: To identify the cortical areas engaged during Chinese word processing using functional magnetic resonance imaging (fMRI) and to examine the reliability and reproducibility of fMRI for localization of functional areas in the human brain. METHODS: FMRI data were collected on 8 young, right-handed, native Chinese speakers during performance of Chinese synonym and homophone judgment tasks on two different clinical MRI systems (1.5 T GE Signa Horizon and 1.5 T Siemens Vision). A cross correlation analysis was used to statistically generate the activation map. RESULTS: Broca's area, Wernicke's area, bilateral extrastriate, and ventral temporal cortex were significantly activated during both the synonym and homophone activities. There was essentially no difference between results acquired on two different MRI systems. CONCLUSIONS: FMRI can be used for localizing cortical areas critical to Chinese language processing in the human brain. The results are reliable and well reproducible across different clinical MRI systems. PMID- 12775225 TI - Validation of diagnostic algorithms for syndromic management of sexually transmitted diseases. AB - OBJECTIVES: To validate our revised syndromic algorithms of the management of sexually transmitted diseases and determine their sensitivity, specificity, positive predictive value and cost-effectiveness. METHODS: Patients with either urethral discharge, vaginal discharge or genital ulcer, were selected during their first visits to three urban sexually transmitted disease clinics in Fujian Province, China. They were managed syndromically according to our revised flowcharts. The etiology of the syndromes was detected by laboratory testing. The data were analyzed using EPI INFO V6.0 software. RESULTS: A total of 736 patients were enrolled into the study. In male patients with urethral discharge, the sensitivities for gonococcal and chlamydial infections were 96.7% and 100%, respectively, using the syndromic approach. The total positive predictive value was 73%. In female patients with vaginal discharge, the sensitivity was 90.8%, specificity 46.9%, positive predictive value 50.9%, and negative predictive value 89.3% for the diagnosis of gonorrhea and/or chlamydial infection by syndromic approach. In patients with genital ulcer, the sensitivities were 78.3% and 75.8%, specificities of 83.6% and 42.9%, and positive predictive values of 60.0% and 41.0% for the diagnosis of syphilis and genital herpes, respectively, using the syndromic approach. Cost-effectiveness analysis indicated that the average cost of treatment for a patient with urethral discharge was RMB 46.03 yuan using syndromic management, in comparison with RMB 149.19 yuan by etiological management. CONCLUSIONS: The syndromic management of urethral discharge was relatively effective and suited clinical application. The specificity and positive predictive value for syndromic management of vaginal discharge are not satisfactory. The revised flowchart of genital ulcer syndrome could be suitable for use in clinical settings. Further validation and revision are needed for syndromic approaches of vaginal discharge and genital ulcer. PMID- 12775226 TI - Prognosis of primary liver carcinoma treated with local resection. AB - OBJECTIVE: To discuss the prognosis of primary liver carcinoma treated with local resection and factors affecting prognosis. METHODS: The data of 130 patients who had been treated from October 1989 to October 1995 were analyzed retrospectively. We analyzed the concrete operating methods, the cause of postoperative complications and factors affecting prognosis. RESULTS: Curative local resection was performed in 130 patients. Operation-related mortality was 0.8% and the incidence of complication was 16.1% (n = 18). The overall 1-, 3- and 5-year survival rates were 82.1%, 60.6% and 48.2%, respectively. Involvement of blood vessels or liver capsules and the elevation of AST before operation were the significant factors affecting prognosis (P < 0.05). CONCLUSION: Local resection is a safe method characterized by little damage, rapid, less blood loss, low complication rate and good prognosis. PMID- 12775227 TI - Use of PCR and PCR-SSP for detection of urinary donor-origin DNA in renal transplant recipients with acute rejection. AB - OBJECTIVE: To analyze the urine of renal recipients for the presence of donor DNA in an attempt to establish an alternative diagnostic means of acute rejection. METHODS: Sixty-four renal transplant recipients were examined. Thirty-seven were normal after transplantation, while 22 others developed acute rejection, based on serum creatinine levels and/or needle biopsy findings of the graft. Five developed drug-induced renal dysfunction. In female recipients with a male graft, we examined urine for the presence of Y chromosome (SRY and DYZ-1) and in recipients receiving an HLA mismatched graft, we looked for HLA-DR gene (DRB1) using PCR. RESULTS: Among the 14 female recipients with male grafts demonstrating stable renal function, only one was positive for SRY and DYZ-1 on the Y chromosome. However, SRY and DYZ-1 were found in the urine of four female patients with acute rejection, but these DNA fragments were not detected in 3 of the 4 after anti-rejection therapy. The last patient was referred to hemodialysis. Of 23 recipients of a graft from HLA mismatch donors with stable renal function, DRB1 was negative in 21 (91%). Of 18 patients with acute rejection, DRB1 was positive in 16 (89%) and negative in 2. These DNA fragments were no longer found in 13 patients after anti-rejection therapy. In all patients with drug induced renal dysfunction, donor-derived DNA was negative. CONCLUSIONS: Presence of door specific DNA in the urine of the recipient is strongly associated with acute rejection. Analysis of DNA derived from donor cells in urine was an effective and accurate method for the diagnosis of acute rejection of a renal transplant. PMID- 12775228 TI - Hypertensive intracranial hematomas: endoscopic-assisted keyhole evacuation and application of patent viewing dissector. AB - OBJECTIVE: To study the effect of endoscopic-assisted keyhole operation (EAKO) on treating hypertensive intracranial hematomas and the value of our patent dissector applied during the operation. METHODS: A total of 25 patients with hypertensive intracranial hematomas underwent endoscopic-assisted keyhole evacuation, during which, the viewing dissector, which had recently achieved national patent, was connected to the tip of endoscope and used to help dissect hematomas. The outcome of this procedure were compared with those of 22 comparable cases undergone conventional surgical treatment (large or smaller craniotomy). The items for comparison included the volume of remaining hematoma, the duration of operation, postsurgical Glasgow Coma Scale (GCS) and Glasgow Outcome Scale (GOS). RESULTS: Remaining hematoma was ascertained 48 h after operation with the use of computerized tomography (CT) scans. In the case of EAKO, nearly complete evacuation (> 84%) was achieved in 21 cases; GCS was evaluated at 7 d postsurgery resulting in GCS > 12 in 9 patients, GCS 9 - 12 in 12 patients and GCS < 9 in 4 patients. The follow-up period ranged from 6 to 21 mon. GOS was estimated at half a year and good recovery rate as defined by GOS was assigned to 76% of the EAKO patients. There are significant differences in the volumes of remaining hematomas and the duration of operation between the EAKO and craniotomy group (P < 0.05). In addition, better clinical outcomes were obtained in EAKO. CONCLUSION: EAKO has the advantage of being minimally invasive, improving surgical results and the prognosis of hypertensive intracranial hematoma patients. We conclude that keyhole operation is a safe, effective alternative for removal of hypertensive intracranial hematoma, particularly during acute stages. PMID- 12775229 TI - Colorectal cancer screening for the natural population of Beijing with sequential fecal occult blood test: a multicenter study. AB - OBJECTIVE: To assess the prevalence of colorectal cancer (CRC) in Beijing and the reliability of the sequential fecal occult blood test (SFOBT) for CRC screening. METHODS: Of the natural population (48 100 persons) in several Beijing communities, we screened 26 827 persons with age over 30 using the SFOBT screening program, Guaiacum Fecal Occult Blood Test (GFOBT), Immuno Fecal Occult Blood Test (IFOBT), and colonoscopies. RESULTS: The screening rate of the population was 74%. The positive rate of SFOBT was 5.6%. The prevalence of CRC in the entire population of Beijing was therefore calculated to be 36.57/10(5). Of 12 CRC detected patients, 4 cases were in stage Dukes A (33.33%), 7 cases in stage Dukes B (58.33%), only 1 case (8.34%) in stage Dukes C. CONCLUSIONS: The prevalence of CRC in Beijing is one of the highest in China. Individuals at high risk for CRC or those over 50 years of age should be considered as primary candidates for screening. SFOBT screening is a cost-effective and reliable method for early detection of CRC. PMID- 12775230 TI - Effect of preoperative transcatheter arterial chemoembolization on apoptosis of hepatocellular carcinoma cells. AB - OBJECTIVE: To evaluate the effect of preoperative transcatheter arterial chemoembolization(TACE) on apoptosis of hepatocellular carcinoma (HCC) cells. METHODS: A total of 136 patients with HCC underwent liver resection. One to five courses of TACE prior to liver resection were performed in 79 patients (TACE group), in which one to four courses of chemotherapy alone were performed in 11 patients (group A), one to five courses of chemotherapy combined with iodized oil were performed in 33 patients (group B), one to three courses of chemotherapy combined with iodized oil and gelatin sponge were performed in 23 patients group C) and one to three courses of chemotherapy combined with iodized oil, ethanol and gelatin sponge were performed in 12 patients (group D). The other 57 patients only received liver resection (non-TACE group). The extent of apoptosis was analyzed by transferase-mediated dUTP nick end labeling (TUNEL) staining. The expressions of Bcl-2 and Bax protein were detected by immunohistochemical method. RESULTS: The apoptotic index(AI) and level of Bax protein in HCC cells were significantly higher in groups A, B, C and D than those in the non-TACE group (P < 0.05). The level of Bcl-2 protein and ratio of Bcl-2 to Bax protein of HCC cells were significantly lower in Groups A, B, C and D than those in the non-TACE group (P < 0.05). CONCLUSION: Preoperative TACE regimens may enhance apoptosis of HCC cells by up-regulating the expression of Bax protein and down-regulating the expression of Bcl-2 protein and ratio of Bcl-2 to Bax protein expression. PMID- 12775231 TI - Clinical features and anesthetic management of multiple endocrine neoplasia associated with pheochromocytoma. AB - OBJECTIVE: To investigate clinical features and anesthetic management of multiple endocrine neoplasia (MEN) associated with pheochromocytoma. METHODS: Medical records of patients who were diagnosed as multiple endocrine neoplasia associated with pheochromocytoma in our hospital from April 1977 to April 2001 were reviewed retrospectively. The demographic data, clinical presentations, family history, biochemical examinations, type of MEN, sequence of different surgical procedures, anesthetic methods and hemodynamics during surgery were analyzed. RESULTS: Thirteen cases of MEN associated with pheochromocytoma were investigated, accounting for 6% (13/213) of the pheochromocytoma patients admitted into our hospital. Nine of the 13 patients presented as type IIa MEN (Sipple syndrome), one as type IIb MEN, and three as mixed MEN. Four patients with type IIa MEN had a family history of similar disease. Five patients with other coexisting endocrine disorders first underwent excision of the pheochromocytomas, although only two had hypertensive symptoms at the time of admittance. Seven patients without histories of hypertension received surgical treatment for pheochromocytoma secondly. The excision of pheochromocytoma was performed under general anesthesia in 8 patients and epidural block in 4 patients. Marked hemodynamic fluctuation was recorded in 8 patients. No perioperative death was recorded. CONCLUSION: Pheochromocytoma may be linked to other endocrine disorders during MEN, either as the main clinical presentation or most frequently as an occult tumor. Recognition of this feature of pheochromocytoma is of importance to the improvement of diagnosis and treatment both for pheochromocytoma and MEN. PMID- 12775232 TI - Use of power Doppler sonography for differential diagnosis of small hepatocellular carcinoma and adenomatous hyperplastic nodule. AB - OBJECTIVE: To evaluate applicability of power Doppler sonography (PDS) in differential diagnosis of small hepatocellular carcinoma (SHCC) and adenomatous hyperplastic nodule (AHN). METHODS: Twenty-two cases of SHCC and 15 cases of AHN were investigated by PDS and the findings were compared with those of digital subtraction angiography (DSA). RESULTS: The rates of arterial and portal flow in an afferent tumor vessel were 86.4% and 40.9% in SHCCs, respectively. The rate of portal flow in an afferent tumor vessel was 60.0% in AHNs, while no arterial flow was detected (P < 0.01). In addition, PDS revealed a constant flow in an efferent tumor vessel (50.0%) continuing to a portal branch in 10 (45.5%) of the 22 SHCCs cases to a hepatic vein in 1 (4.5%) of the 22 SHCCs, but to nothing else in the AHNs (P < 0.01). CONCLUSIONS: Power Doppler sonography is of value in distinguishing SHCC from AHN, and arterial afferent tumor vessels from constant flow efferent tumor vessels at PDS. PMID- 12775233 TI - Effect of microsomal triglyceride transfer protein gene polymorphism in the promoter region on dyslipidemia in type 2 diabetic subjects. AB - OBJECTIVE: To explore the relationship between microsomal triglyceride transfer protein (MTP) gene variation and diabetic dyslipidemia among Chinese. METHODS: Using PCR restriction fragment length polymorphism (PCR-RFLP) analysis and gene sequencing, we studied the influence of a common MTP gene polymorphism in the promoter region on the apoB-containing lipoproteins in 44 Chinese type 2 diabetic subjects and 32 non-diabetic volunteers. RESULTS: A common functional G/T polymorphism in 493 bp upstream from the transcriptional start point was detected among native Chinese. There were 41 carriers (53.9%) of the MTP-493 G/G genotype, 28 (36.8%) of the MTP-493 G/T genotype and 7 (9.3%) of the MTP-493 T/T genotype. The allele frequency of MTP-493 T in the diabetic group was 0.30. The MTP-493 T/T diabetic group had significantly higher TG (P < 0.05), VLDL-CH (P < 0.05) and smaller LDL particle size (P < 0.001) than the MTP-493 common genotype group. CONCLUSION: Genetic variation in the MTP promoter is likely to be highly involved in the production of dyslipidemia in type 2 diabetic subjects. PMID- 12775234 TI - Expression and significance of IL-18 in the bone marrow of patients with hematological diseases. AB - OBJECTIVE: To investigate the levels of IL-18 in the bone marrow of both normal subjects and patients with hematological diseases and to determine the possible significance of IL-18 in pathogenesis of some hematological malignancies. METHODS: The IL-18 mRNA levels in the bone marrow of 140 patients with hematological diseases and 15 normal donors were determined by the semi quantitative reverse transcriptase polymerase chain reaction (RT-PCR). Immunohistochemical method was used to detect IL-18 protein in 12 patients with acute myeloid leukemia (AML). The possible regulation of IL-18 for proliferation of some leukemia cells was investigated using antisense techniques. RESULTS: IL 18 mRNA levels were obviously higher in the patients with leukemia or other malignant hematological diseases (OMHD) than in normal donors. However, no significant difference was found in the level of transcription between patients with iron deficiency anemia (IDA) and normal controls. Immunohistochemical method confirmed the presence of IL-18 protein in 10 out of 12 AML cases with positive transcription. By 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, IL-18 antisense oligodeoxynucleotides (ASON) clearly inhibited the growth of J6-1 and HL-60 cells (42% and 12% inhibited, respectively) in a dose dependent manner. CONCLUSIONS: IL-18 was detected at elevated levels in the bone marrow of patients with some hematological malignancies, and might be involved in the proliferation of certain leukemic cells in vivo through an autocrine mechanism. PMID- 12775235 TI - Analysis of long-term outcomes of the posterior interosseous flap and compound flap. AB - OBJECTIVE: To analyze the long-term outcomes of antebrachial flap using a pedicle of the dorsal interosseous artery. METHODS: Ninety cases were followed up for 1 16 years, in which the indications, long-term outcomes and dorsal aspect of the forearm were assessed retrospectively. RESULTS: Eighty-nine cases survived with satisfactory contour of the forearm and hand. CONCLUSIONS: Antebrachial flap using a pedicle of the dorsal interosseous artery is an optimal method for repair of soft tissue defects in the hand. PMID- 12775236 TI - Treatment of 522 patients with Nevus of Ota with Q-switched Alexandrite laser. AB - OBJECTIVE: To evaluate the clinical response of Nevus of Ota to Q-switched Alexandrite laser, and analyze factors that influence the treatment outcome. METHODS: A total of 522 patients treated with Q-switched Alexandrite laser were included in the study. Single and multiple variate analyses of various factors were performed. RESULTS: Satisfactory result was observed in all patients, none of whom developed scarring. Clinical response was improved with additional treatment sessions. The clinical response of the 20 - 27-week treatment interval group was significantly better than that of the 12 - 19-week interval group, but showed no significant difference as compared with both the 28 - 35 and > or = 36 week interval group. Zygomatic, buccal and frontal areas showed better response than ocular and temporal areas. Treatment session, interval, and fluence were significant factors identified by multivariate analysis. CONCLUSIONS: Q-switched Alexandrite laser is an ideal method for treating Nevus of Ota without injury. The number of treatment sessions is more important than interval or fluence. PMID- 12775237 TI - Clinical, cytogenetic and dual-color FISH studies on five cases of myelodysplastic syndrome or acute myeloid leukemia patients with 1;7 translocation. AB - OBJECTIVE: To study the clinical and cytogenetic characteristics of four patients with myelodysplastic syndrome (MDS) and one with acute myeloid leukemia experiencing t (1;7). METHODS: Five patients seen in our hospital from 1992 to 2001 were diagnosed as MDS and acute myelocytic leukemia (AML) according to the French-American-British (FAB) criteria. Chromosomes were prepared using the direct method as well as 24-hour unstimulated cultures of fresh heparinized bone marrow for each subject, while R-banding was used to analyze karyotypes. Dual color fluorescence in situ hybridization (FISH) using SpectrumRed and SpectrumGreen directly labeled chromosome 1-specific alpha-satellite DNA probe (red) and chromosome 7- specific alpha-satellite DNA probe (green) was performed for three cases. RESULTS: Of the five patients, three had 1;7 translocation due to a long history of exposure to benzene. In three cases, dual-color FISH resulted in three red signals and two green ones, in which one red signal adjoining one green signal in 27.6%, 84% and 18.5% metaphases, respectively. CONCLUSIONS: Exposure to benzene may be the cause for Chinese MDS and AML patients with t (1;7) translocation. The result of dual-color FISH convincingly confirmed that the centromere of the derivative chromosome 7p/1q resulting from 1;7 translocation was made up of centromeres from both chromosomes 1 and 7. PMID- 12775238 TI - Thermal injuries induce gene expression of endogenous c-fos, c-myc and bFGF in burned tissues. AB - OBJECTIVE: To investigate the expression sequence and distribution characteristics of the protooncogenes c-fos, c-myc and endogenous basic fibroblast growth factor (bFGF) genes in burned tissues, and to explore the possible effects of changes in these genes' functions on wound healing. METHODS: Partial-thickness burns of 30% TBSA were established on backs of Wistar rats. In situ hybridization and histological methods were used to detect expression of c fos, c-myc and bFGF genes in normal and burned tissue at 3 h, 6 h, 1 d, 3 d, 7 d and 14 d postburn. RESULTS: Although expression of c-fos and c-myc genes and bFGF gene could be found in normal skin, the expression of all three were markedly induced by burn wounds and the expression models in sequence and distribution were quite different. Expression of c-fos gene increased and peaked at 6 h. Signals were mainly localized in both nuclei of dermal fibroblasts and monocytes. The expression of bFGF gene increased at 6 h and peaked at 1 d postburn, and was distributed in the cytoplasm of fibroblasts. C-myc gene peaked 3 d postburn and was also distributed in the cytoplasm of fibroblasts. CONCLUSIONS: These results indicated that thermal injury could induce the expression of c-fos, c-myc and bFGF at gene level, showing phasic control and regional distribution. The phasic expression of these genes suggests that there is an interaction between protooncogenes and bFGF, which may play an important role in wound healing. The different expressions of c-fos and c-myc play an inducing role in regulating bFGF, and in turn affect wound healing. PMID- 12775239 TI - Characterization of Sporothrix schenckii by random amplification of polymorphic DNA assay. AB - OBJECTIVES: To investigate the DNA polymorphism of Sporothrix schenckii (S. schenckii) and to find the relationship between DNA patterns and geographic areas and clinical manifestations. METHOD: The total DNA was extracted with hexadecyltrimethyl-ammonium bromide. Random Amplification of Polymorphic DNA (RAPD) assay was used to study DNA typing of 24 strains of S. schenckii collected from different areas and isolated from different clinical types. RESULTS: Of seven random primers used, three primers (OPAA11, OPD18 and OPB07) gave good reactions, the sequences of which were 5'-ACCCGACCTG-3', 5'-GAGAGCCAAC-3', 5' GGTGAC~GCAG-3' respectively. The RAPD patterns of the 24 isolates were not completely identical, showing certain degrees of hereditary variability. Different isolates showed a common conserved DNA band with the same primer. Different clinical types showed different genotypes. CONCLUSION: RAPD analysis is useful in DNA typing of S. schenckii, the DNA band type of which is related to geographic origin and Clinical manifestation. PMID- 12775240 TI - Local tissue hypoxia and formation of nasal polyps. AB - OBJECTIVE: To explore the response of nasal mucosa epithelial cells to hypoxia in terms of formation of nasal polyps (NP). METHODS: Epithelial cells of NP and inferior turbinate (IT) were cultured serum-free under normal oxygen and hypoxic circumstances with stimulation of IL-1 beta and TNF alpha. The vascular endothelial growth factor (VEGF) mRNA and VEGF protein levels of the cultured cells were detected using in situ hybridization and ELISA, respectively. RESULTS: The expression of VEGF mRNA was significantly higher in epithelial cells of NP than in IT exposed to pro-inflammatory cytokines or hypoxia (P < 0.01). VEGF levels were higher in NP epithelial cells than those of IT (P < 0.01) under hypoxia. CONCLUSION: VEGF-induced by hypoxia is very important for the early stages of forming polyps. PMID- 12775241 TI - Therapeutic effect of AdCMVCD/5-FC system and metabolism of 5-FC in the treatment of human tongue squamous cell carcinoma. AB - OBJECTIVE: To investigate the therapeutic effect and metabolism of 5 fluorocytosine (5-FC) in human tongue squamous carcinoma cells after treatment with adenovirus-mediated cytosine deaminase (AdCMVCD)/5-FC system. METHODS: Human tongue squamous carcinoma cells (Tca8113 cell line) and its xenografts in BALB/c nude mice were treated with AdCMVCD/5-FC system. The killing effect in vitro and bystander effect were detected by microculture tetrazolium (MTT) assay. Tumor inhibition effect and histopathological changes were observed in vivo. High performance liquid chromatography (HPLC) was performed to determine the metabolism of 5-FC in vitro and in vivo. RESULTS: AdCMVCD/5-FC system had strong killing effect and bystander effect on Tca8113 cells. Both condition media and cell extracts showed two peaks identified as 5-FC and 5-fluorouracil (5-FU) by HPLC and a time-dependent generation of 5-FU and concomitant time-dependent decreases of 5-FC. Compared to the control groups, mice treated with AdCMVCD/5-FC system demonstrated significant tumor regression (P < 0.001); the tumor doubling time prolonged and inhibition rate was 92.62%. There were substantial tumor necrotic areas and infiltrative lymphocytes around necrotic areas in the AdCMVCD/5-FC treated group under light microscope. There was a significantly low concentration of 5-FC and high concentration of 5-FU in tumor tissue, but only 5 FC was found in blood. CONCLUSION: AdCMVCD/5-FC suicide gene system had significant in vitro and in vivo anti-tumor effect on human tongue squamous cell carcinoma due to convert 5-FC into 5-FU. PMID- 12775242 TI - Ischemic preconditioning in immature hearts: mechanism and compatibility with cardioplegia. AB - OBJECTIVE: To investigate (1) whether ischemic preconditioning (IPC) could protect immature rabbit hearts against ischemia-reperfusion injury and (2) the role of K(ATP) channel in the mechanism of myocardial protection. Since cardioplegia is a traditional and effective cardioprotective measure in clinic, our study is also designed to probe the compatibility between IPC and cardioplegia. METHODS: New Zealand rabbits aged 14 - 21 days weighing 220 - 280 g were used. The animals were anesthetized and heparinized. The chest was opened and the heart was quickly removed for connection of the aorta via Langendorff's method within 30 s after excision. All hearts were perfused with Krebs-Henseleit buffer balanced with gas mixture (O(2):CO(2) = 95%:5%) at 60 cm H(2)O (perfusion pressure). IPC consisted of 5 min global ischemia plus 10 min reperfusion. Glibenclamide was used as the K(ATP) channel blocker at a concentration of 10 micro mol/L before IPC. Cardiac arrest was induced with 4 degrees C St. Thomas cardioplegic solution, at which point the heart was made globally ischemic by withholding perfusion for 45 min followed by 40 min reperfusion. Thirty immature rabbit hearts were randomly divided into four groups: CON (n = 9) was subjected to ischemia-reperfusion only; IPC (n = 9) underwent IPC and ischemia-reperfusion; Gli (n = 6) was given glibenclamide and ischemia-reperfusion; and Gli + IPC (n = 6) underwent glibenclamide, IPC and ischemia-reperfusion. Coronary flow (CF), HR, left ventricle developed pressure (LVDP), and +/- dp/dt(max) were monitored at equilibration (baseline value) and 5, 10, 20, 30 and 40 min after reperfusion. The values resulting from reperfusion were expressed as a percentage of their baseline values. Arrhythmia quantification, myocardial enzyme in the coronary effluent and myocardial energy metabolism were also determined. RESULTS: The recovery of CF, HR, LVDP and +/- dp/dt(max) in preconditioned hearts was best among the four groups. The incidence of arrhythmia was low and less CK-MB leaked out in the IPC group. Myocardial ATP content was better preserved by IPC. Pretreatment with glibenclamide completely abolished the myocardial protection provided by IPC, but did not affect ischemia-reperfusion injury. CONCLUSIONS: While applying cardioplegia, IPC provides significant cardioprotective effects. Activation of K(ATP) channels is involved in the mechanism of IPC-produced cardioprotection. PMID- 12775243 TI - Nitric oxide opens second window of protection in ischemic preconditioning via induction of heat-shock protein 72. AB - OBJECTIVE: To examine the inhibition of nitric oxide (NO) synthesis during ischemic preconditioning (IP) upon the induction of heat-shock protein 72 (HSP72) and the size-limiting effect of the second window of protection on infarction. METHODS: Rabbits were subjected to either 4 cycles of 5-min long coronary artery occlusion separated by 10 min of reperfusion, or a sham operation. During this procedure, we administered 10 mg/kg of N(G)-nitro-L-arginine methyl ester (L NAME, an inhibitor of NO synthase) intravenously 5 min before IP followed by its continuous infusion (1.5 mg/kg/min). Twenty-four hours after IP or the sham operation, the hearts were rapidly excised for assay of HSP72 expression or were subjected to 30 min of coronary artery occlusion followed by 120 min of reperfusion, at which point infarct size (IS) was measured. RESULTS: Twenty-four hours after IP or the sham operation, there was no difference in heart rate or mean arterial pressure between the groups. Immunoblotting revealed an increase in HSP72 protein levels in the IP group, which was blocked by L-NAME. IS in the IP rabbits was reduced compared with controls (29.8 +/- 3.7% vs. 50.8 +/- 4.3%, P < 0.01). IS in the IP rabbits was elevated as a result of L-NAME treatment (46.0 +/ 5.1%). Administration of L-arginine reversed the effects of L-NAME on the induction of HSP72 and IS (33.5 +/- 4.0%). The intravenous administration of S nitroso-N-acetylpenicillamine (an NO donor, 15 microg/kg/min) over 20 min increased the induction of HSP72 and reduced IS (31.3 +/- 5.7%, P < 0.01 vs. control) 24 h later. CONCLUSION: These findings suggest that NO may be involved in the induction of HSP72 and the opening of the second window of protection of IP. PMID- 12775244 TI - Relationship between coronary arterial remodeling and clinical presentation. AB - OBJECTIVE: To examine the relationship between coronary arterial remodeling and clinical presentation. METHODS: A total of 34 patients with acute (10 with recent myocardial infarction and 24 with unstable angina) and 26 with stable (8 with old myocardial infarction and 18 with stable angina) coronary syndrome underwent intravascular ultrasound (IVUS) before intervention. Target lesions were classified as soft or hard plaques. Quantitative measurements of cross-sectional area (CSA) of external elastic membrane (EEM), lumen and plaque were performed at the lesion site and at the proximal and distal reference sites. Remodeling index (RI) was expressed by the ratio of EEM CSA at the lesion site to the mean EEM CSA of both proximal and distal reference sites. Positive remodeling was defined as RI > 1.05 and negative remodeling as RI < 0.95. RESULTS: Soft plaque was observed more frequently in acute than in stable coronary syndrome (59% vs 31%), whereas hard plaque was more common in stable coronary syndrome (69% vs 41%) (P = 0.03). The EEM CSA (15.11 +/- 2.89 mm(2) vs 13.25 +/- 3.10 mm(2), P = 0.019) and plaque CSA (10.83 +/- 2.62 mm(2) vs 9.30 +/- 2.84 mm(2), P = 0.035) were significantly greater at target lesions in patients with acute rather than stable coronary syndrome, while lumen CSA and percent area stenosis were similar in both groups. RI was significantly higher (1.08 +/- 0.16 vs 0.95 +/- 0.14, P = 0.002) and positive remodeling was more frequent in acute coronary syndrome (53% vs 23%, P = 0.019), whereas negative remodeling was more common in stable coronary syndrome (58% vs 24%, P = 0.007). CONCLUSIONS: The study indicates that clinical characteristics of patients with coronary artery disease depend largely upon underlying types of coronary arterial remodeling. PMID- 12775245 TI - Labeling embryonic stem cells with enhanced green fluorescent protein on the hypoxanthineguanine phosphoribosyl transferase locus. AB - OBJECTIVE: To label embryonic stem (ES) cells with enhanced green fluorescent protein (EGFP) on the hypoxanthineguanine phosphoribosyl transferase (HPRT) gene locus for the first time to provide a convenient and efficient way for cell tracking and manipulation in the studies of transplantation and stem cell therapy. METHODS: Homologous fragments were obtained by polymerase chain reaction (PCR), from which the gene targeting vector pHPRT-EGFP was constructed. The linearized vector was introduced into ES cells by electroporation. The G418(r)6TG(r) cell clones were obtained after selection with G418 and 6TG media. The integration patterns of these resistant cell clones were identified with Southern blotting. RESULTS: EGFP expressing ES cells on the locus of HPRT were successfully generated. They have normal properties, such as karyotype, viability and differentiation ability. The green fluorescence of EGFP expressing cells was maintained in propagation of the ES cells for more than 30 passages and in differentiated cells. Cultured in suspension, the "green" ES cells aggregated and formed embryoid bodies, retaining the green fluorescence at varying developmental stages. The "green" embryoid bodies could expand and differentiate into various types of cells, exhibiting ubiquitous green fluorescence. CONCLUSIONS: This generation of "green" targeted ES cells is described in an efficient protocol for obtaining the homologous fragments by PCR. Introducing the marker gene in the genome of ES cells, we should be able to manipulate them in vitro and use them as vehicles in cell-replacement therapy as well as for other biomedical and research purposes. PMID- 12775246 TI - Role of adhesion molecules in mobilization of hematopoietic cells. AB - OBJECTIVE: To study the changes of adhesion molecules' expressions during the recombinant human granulocyte-colony-stimulating factor (rhG-CSF) mobilization in peripheral blood stem cell transplantation (PBSCT), and to confirm the influence of rhG-CSF on hematopoietic stem cells, which are proposed to guide mobilization in PBSCT. METHODS: Mice were injected subcutaneously with diluted rhG-CSF or normal saline for 7 days. The blood Sca-1(+) stem cell count and bone marrow (BM) nucleated cell count were enumerated. The expressions of CD49d and CD44 and the adhesive ability of mononuclear cells to bone marrow matrix (fibronectin) were examined by flow cytometry and (51)Cr adhesive assay, respectively. RESULTS: The mobilizing effect of rhG-CSF on mice was the same as on humans. The number of Sca 1(+) cells in peripheral blood reached the peak on the seventh day, the BM nucleated cell count was reduced, and the expressions of CD49d and the cells' adhesive ability in BM and PB declined. CONCLUSIONS: rhG-CSF can reduce some cell adhesion molecules' expressions and the adhesive ability of hematopoietic stem cells to BM matrix, therefore mobilizing hematopoietic stem cells (HSC) from the BM to the peripheral blood. PMID- 12775247 TI - Cardiac conductive system excitation maps using intracardiac tissue Doppler imaging. AB - OBJECTIVE: To precisely visualize cardiac anatomic structures and simultaneously depict electro-mechanical events for the purpose of precise underblood intervention. METHODS: Intracardiac high-resolution tissue Doppler imaging was used to map real time myocardial contractions in response to electrical activation within the anatomic structure of the cardiac conductive system using a canine open-chest model. RESULTS: The detailed inner anatomic structure of the cardiac conductive system at different sites (i.e., sino-atrial, atrial wall, atrial-ventricular node and ventricular wall) with the inside onset and propagation of myocardial velocity and acceleration induced by electrical activation was clearly visualized and quantitatively evaluated. CONCLUSION: The simultaneous single modality visualization of the anatomy, function and electrical events of the cardiac conductive system will foster target pacing and precision ablation. PMID- 12775248 TI - Effects of oral administration of type II collagen on adjuvant arthritis in rats and its mechanisms. AB - OBJECTIVE: To investigate the effects of oral administration of type II collagen (CII) on adjuvant arthritis (AA) in rats and its mechanisms, and to compare the effects of CII with those of the Chinese traditional medicine Tripterygium Polyglycoside administered similarly. METHODS: Arthritis was induced in rats by immunization using Freund's complete adjuvant (FCA). After feeding rats either soluble CII or Tripterygium Polyglycoside, changes in degree of articular swelling and articular histological findings were observed in AA rats. Some correlative immunological indexes were measured, including delayed type hypersensitivity (DTH) reaction, anti-collagen and anti-Mycobacterium tuberculosis (MT) antibody in serum, and levels of IFN-gamma and TNF-alpha in articular steep in rats. RESULTS: Oral administration of CII was able to alleviate both distinctly articular and general symptoms in AA rats, suppress synovium hyperplasia and inflammatory cells infiltration in arthrosis capsule. The effects brought about by CII were stronger than those by Tripterygium Polyglycoside. Oral administration of CII inhibited antigen-specific immune response, such as DTH and antibody reaction to CII. In addition, the expression of IFN-gamma and TNF-alpha in joints were locally downregulated. CONCLUSIONS: The therapeutic effect of oral administration of CII is obvious on adjuvant arthritis in rats. Its remedial mechanisms are likely related to the downregulation of both IFN-gamma and TNF-alpha, and the suppression of cell immunity. PMID- 12775249 TI - Therapeutic efficacy of tumor-derived heat shock protein 70 immunotherapy combining interleukin-2 on tumor-bearing mice. AB - OBJECTIVE: To investigate the therapeutic efficacy of compound immunotherapy of tumor-derived heat shock protein 70 (HSP70) and interleukin-2 (IL-2) on tumor bearing mice, and to provide reference for translating this strategy to human cancer. METHODS: Cell culture, techniques for protein extraction and purification, SDS-PAGE, Western blot and capillary electrophoresis for HSP70 detection and purity analysis, and animal experiments were used. Mice were treated with HSP70 5 or 10 microg and IL-2 50 kU, 100 kU or 2 kU (maintaining dosage) at previously designated intervals. RESULTS: Both the mono-administration of either HSP70 or IL-2 and the compound immunotherapy of HSP70 and IL-2 obviously inhibited the growth of the implanted tumor and prolonged the life span of the mice to different extents. However, long periods of tumor-free survival (over 90 days) were demonstrated only in HSP70 10 micro g group, HSP70 10 microg IL-2 50 kU group, and HSP70 10 microg-IL-2 100 kU group (40%, 40%, 60% respectively). On the other hand, none of the mice in the rest groups achieved long-term survival. Statistical significance was apparent in comparison with the groups without long period survival (P < 0.025 - 0.05). CONCLUSION: Our research revealed that tumor-derived HSP70 immunotherapy was much more effective than IL-2 alone. And in compound immunotherapy, HSP70 was the main factor in delaying or eradicating the tumors. The proper combination of HSP70 and IL-2 (10 microg HSP70 and 100 kU IL-2 in this experimental mouse model) clearly enhanced the immunotherapy efficacy which indicated that the specific immunotherapy as a main part of tumor immunotherapy assisted by cytokine immunotherapy would be a promising strategy in cancer treatment. PMID- 12775250 TI - Effects of propofol, midazolam and thiopental sodium on outcome and amino acids accumulation in focal cerebral ischemia-reperfusion in rats. AB - OBJECTIVE: To investigate the effects of propofol, midazolam and thiopental sodium on outcomes and amino acid accumulation in focal cerebral ischemia reperfusion in rats. METHODS: Male Sprague Dawley (SD) rats were scheduled to undergo 3-hour middle cerebral artery occlusion by intraluminal suture and 24 hour reperfusion. Neurologic outcomes were scored on a 0-5 grading scale. Infarct volume was shown with triphenyltetrazolium chloride staining and measured by an image analysis system. Concentrations of various amino acids (aspartate, glutamate, glycine, taurine, and gama-aminobutyric acid) were measured after 3 hours of reperfusion using high performance liquid chromatography. Propofol, midazolam and thiopental sodium were given intraperitoneally at the beginning of reperfusion. RESULTS: Both propofol and midazolam attenuated neurological deficits and reduced infarct and edema volumes. Propofol showed better neurological protection than midazolam while thiopental sodium did not exhibit any protective effect. Both propofol and midazolam decreased excitatory amino acids accumulation, while propofol increased gama-aminobutyric acid accumulation in ischemic areas in reperfusion. CONCLUSION: Propofol and midazolam, but not thiopental sodium, may provide protective effects against reperfusion induced injury in rats subjected to focal cerebral ischemia. This neurological protection may be due to the acceleration of excitatory amino acids elimination in reperfusion. PMID- 12775252 TI - The influence of insulin on secretion of IGF-I and IGFBP-I in cultures of human endometrial stromal cells. AB - OBJECTIVES: To study the influence of insulin on IGF-I and IGFBP-I secretion of the human endometrial stromal cells. METHODS: Late proliferative phase endometrial stromal cells were isolated from endometrium tissues and then cultured for 24 h in Hams F-12 only as a control and in Hams F-12 with different concentrations of estradiol (E2) and insulin (INS) as treated groups. Simultaneously, the endometrial stromal cells from late secretory phase endometrium were cultured for 24 h in Hams F-12 only as a control and in Hams F 12 supplemented with different concentrations of progesterone (P) and insulin as treated groups. After 24 h of culturing, the mediums were collected for either IGF-I or IGFBP-I assays. RESULT: The concentrations of IGF-I in medium from cultured endometrial stromal cells in the proliferative phase were 0.78 +/- 0.47 ng/ml in the hormone-free control group; 1.44 +/- 0.59 ng/ml and 1.39 +/- 0.33 ng/ml in 100 pg/ml E2 group and 20 microU/ml INS group, which was higher than that of the control group (P < 0.05 and P < 0.01, respectively). The IGF-I concentration in the 100 microU/ml INS group was 2.03 +/- 0.53 ng/ml, which was higher than that of the 20 micro U/ml INS group (P < 0.01). Levels of IGF-I in the 100 pg/ml E2 plus 20 microU/ml INS group was 2.18 +/- 0.36 ng/ml, which was significantly higher than that of the 20 microU/ml INS and 100 pg/ml E2 group (P < 0.01), but lower than that of the 100 pg/ml E2 plus 100 microU/ml INS group (3.42 +/- 0.75 ng/ml), P < 0.01. The concentration of IGFBP-I in medium from cultured endometrial stromal cells in the secretory phase was 2.50 +/- 1.39 ng/ml in the hormone-free control group and 5.44 +/- 2.09 ng/ml in the 10 pg/ml P group, which was significantly higher than that of the control (P < 0.01). IGFBP I concentration in 20 microU/ml INS group was 0.16 +/- 0.58 ng/ml, which was lower compared with control, but higher compared with the 100 microU/ml INS group (P < 0.01). The level of IGFBP-I in the 10 ng/ml P plus 20 microU/ml INS group was 2.10 +/- 1.17 ng/ml, lower compared with the 10 ng/ml P group, but higher compared with the 10 pg/ml P plus 100 microU/ml INS group, P < 0.01. CONCLUSIONS: Insulin can stimulate basal (without hormone) and E2-stimulated IGF-I secretion in cultured stromal cells from human late proliferative endometrium in a dose dependent manner. Insulin can suppress basal (without hormone) and P-stimulated IGFBP-I secretions in cultured stromal cells from human secretory endometrium in a dose-dependent manner. PMID- 12775251 TI - Synthesis of Toll-like receptor 4 in Kupffer cells and its role in alcohol induced liver disease. AB - OBJECTIVES: To observe the synthesis of Toll-like receptor (TLR) 4 protein and its mRNA expression in Kupffer cells (KCs) and evaluate the role of TLR 4 in liver injury to rats through alcohol-induced liver disease. METHODS: Twenty-eight Wistar rats were divided into two groups: ethanol-fed (group E) and control (group C). Group E rats were given ethanol at a dose of 5 - 12 g x kg(-1) x d( 1), while group C received dextrose. Animals from both groups were killed at 4 and 8 weeks. The KCs were isolated and synthesis of TLR 4 protein was determined by laser scanning confocal microscopy. TLR 4 mRNA expression in KCs was determined by reverse transcription polymerase chain reaction (RT-PCR) analysis. The levels of endotoxin, tumor necrosis factor-alpha (TNF-alpha) and interleukin 6 (IL-6) in plasma were determined. Changes in liver pathology were observed. RESULTS: Laser scanning confocal microscopy showed that the intensity of fluorescence of TLR 4 protein in group E was stronger than group C. Ethanol administration led to a significant increase in TLR 4 mRNA expression in group E compared with group C (P < 0.05). The concentrations of plasma endotoxin, TNF alpha and IL-6 were higher in group E than in group C (P < 0.05). Liver sections from rats in group E demonstrated marked pathological changes. CONCLUSION: Ethanol administration can lead to the synthesis of TLR 4 protein and its gene expression in KCs, indicating that TLR 4 may play a major role in the development of alcohol-induced liver injury. PMID- 12775253 TI - Relapse risk assessment of transplantation for patients with chronic myeloid leukaemia. AB - OBJECTIVE: To analyse the risk factors of relapse before bone marrow transplantation (BMT) and to present the prognostic information as good as possible. METHODS: A total of 3142 patients, who underwent the allogeneic blood or bone marrow transplantation between 1989 and 1997 and were documented in the European Group for Blood and Marrow transplantation (EBMT), were included. Six possible risk factors including type of donor, stage of disease, age, gender, donor-recipient sex combination and the waiting time from diagnosis to transplantation of relapse were considered. The time to relapse was analysed by Kaplan-Meier curves and Cox regression with stratification on prognostic factors that did not satisfy the Proportional Hazard Assumption. RESULTS: An amount of 447 patients relapsed out of all 3142 patients. The relapse rate was 14.2%. Type of donor and stage of disease showed a clear prognostic effect, but failed the proportional hazard assumption. Therefore, the data were stratified on the combination of type of donor and stage of disease. Within these strata an additional significant effect of age could be observed. Relative risk of age > or = 40 vs age < 40 was 1.32 (95% confidence interval 1.09 - 1.59). The prognostic model is summarized graphically. CONCLUSIONS: The combination of type of donor, stage of disease and age of recipient at transplantation are important prognostic factors for relapse after BMT. PMID- 12775254 TI - Tibetan patients with essential hypertension caused by underlying oxidative metabolism dysfunction and depressed nitric oxide synthesis. AB - OBJECTIVE: To assess the role of oxidative metabolism and nitric oxide synthesis for elucidating their pathophysiological mechanisms in a Tibetan patient with essential hypertension. METHODS: The serum levels of total superoxide dismutase (T-SOD), malondialdehyde (MDA), total antioxidant capacity (T-AOC), nitric oxide (NO) and nitric oxide synthase (NOS) were assayed in sixty native Tibetans (thirty hypertensive patients and thirty healthy volunteers as control). RESULTS: The levels of T-SOD, T-AOC, NO and NOS were significantly lower in the patient group than in the control group (P < 0.01); MDA was significantly higher in the patient group than in the control group (P < 0.01). The level of MDA had a strong negative correlation with T-SOD, T-AOC, NO and NOS (r = -0.82, -0.76, -0.79, 0.73, respectively, P < 0.001 for all). CONCLUSION: Tibetan patients with essential hypertension (EH) may have underlying oxidative metabolism dysfunction and depressed NO synthesis, both responsible for the hypertensive process. PMID- 12775255 TI - An Ile93Met substitution in the UCH-L1 gene is not a disease-causing mutation for idiopathic Parkinson's disease. AB - OBJECTIVE: To ascertain whether a coding mutation (Ile93Met) in ubiquitin carboxy terminal hydrolase (UCH-L1) gene plays a role in idiopathic Parkinson's disease (IPD). METHODS: Polymerase chain reaction-restriction fragment length polymorphism assay (PCR-RFLP) was used to distinguish the wild-type (two DNA fragments of 34 and 126 bp) from the variant allele (three fragments of 34, 60 and 66 bp) because the mutation created a new site for restriction endonuclease BsmF1. DNA was isolated from various blood samples using a phenolchloroform extraction. RESULTS: Ile93Met substitution was found neither in PD patients nor in controls. CONCLUSIONS: Our study suggested that Ile93Met of UCH-L1 gene did not influence risk of IPD. PMID- 12775257 TI - Idiopathic hepatic arterial malformation: a case report. PMID- 12775256 TI - Role of HLA-DR and CD1a molecules in pathogenesis of hypertrophic scarring and keloids. PMID- 12775258 TI - Hepatic angiosarcoma: CT findings. PMID- 12775259 TI - [Primary pulmonary schwannoma: clinical analysis of 7 cases and review of the literature]. AB - OBJECTIVE: To analyze the clinical and radiological manifestations of primary pulmonary schwannoma in order to improve the diagnosis of this rare disease. METHODS: Seven cases of primary pulmonary schwannoma which was confirmed by pathology of surgical specimens were retrospectively analyzed. RESULTS: Three cases were benign (1 of pulmonary parenchymal origin and 2 of bronchial origin) and 4 were malignant (3 of pulmonary parenchymal origin and 1 of bronchial origin). There were four male and three female patients, age ranging from 31 to 74 years. All the 7 cases had chest X-ray films and 6 cases had chest CT examinations. The major complains included slight hemoptysis, cough, fever, chest pain and shortness of breath. Bronchoscopy showed bronchial tumors in 3 cases and partial stenosis of bronchus in 2 cases. Radiology showed nodules in the left main bronchus in 2 cases. Solitary mass was found in 4 cases and multiple masses (2 lesions) were found in one patient. The margin of the lesions was smooth in 3 cases (one malignant and two benign) and irregular in another three cases (all malignant). Chest CT showed reticular enhancement in one case (benign), heterogeneous enhancement in 4 cases (3 malignant and 1 benign), and peripheral enhancement with a large central hypodense area in one case (malignant). Compression of the trachea and ribs was prominent in 2 patients. Pleural effusion and blood vessel infiltration were evident in 3 patients with malignant schwannoma. CONCLUSION: Primary pulmonary schwannoma is rare. The symptoms and CT manifestations are nonspecific. The size and the density of the tumor are not helpful in differentiating malignant schwannoma from the benign one. Marginal irregularity and invasion of adjacent structures are suggestive of malignancy. PMID- 12775260 TI - [Diagnosis of pulmonary sclerosing hemangioma with incremental dynamic CT: analysis of 20 cases]. AB - OBJECTIVE: To evaluate the value of incremental dynamic CT in the diagnosis of pulmonary sclerosing hemangioma (sclerotic hemangioma). METHODS: Thin-section CT at 2.0 mm thickness and 2.0 mm interval was performed before and after administration of contrast material in 20 cases of pulmonary sclerosing hemangioma. RESULTS: Pulmonary sclerosing hemangiomas were shown to be round or oval masses, with smooth margins, homogeneous parenchymal density, and occasional calcification. Homogeneous enhancement was evident in all cases, with maximum CT values ranging from 90 to 110 HU. CONCLUSION: Spiral dynamic CT is useful in the diagnosing of sclerosing hemangioma in the lungs. PMID- 12775261 TI - [FDG dual-head tomography with coincidence imaging and serum tumor marker assays for the assessment of malignancy of pulmonary lesions]. AB - OBJECTIVE: To evaluate the performance of FDG dual-head tomography with coincidence (DHTC) imaging and serum tumor marker assays in identifying lung carcinoma in 160 patients with abnormal findings on chest radiography. METHODS: A prospective evaluation of FDG imaging with DHTC and the measurement of 3 serum tumor markers for lung cancer (carcinoembryonic antigen, CYFRA21-1 and neuron specific enolase) were performed in two weeks in 160 consecutive patients with known or suspected lung malignancy. All images were analyzed visually, and the count ratio of tumor to normal tissue (T/N ratio) was calculated. It was considered positive if the FDG uptake was increased relative to that in the adjacent lung tissue, and the uptake was focal and the T/N ratio > or = 1.3. The serum tumor marker test was considered positive for malignancy if the serum level of at least 1 marker was elevated. RESULTS: 127 patients were proven to have lung cancer by pathology, and 33 patients had benign lung disease. The sensitivity, specificity and accuracy of FDG DHTC in assessing lung cancer were 94.5%, 84.8% and 92.5%, respectively, and those of the serum tumor markers were 67.7%, 84.8% and 71.3%, respectively. FDG DHTC showed significantly higher sensitivity (P < 0.001) and accuracy (P < 0.001) than serum tumor markers. Four patients with lung cancer had negative findings on FDG DHTC but had positive serum markers. CONCLUSION: FDG DHTC imaging is a powerful tool for evaluating patients with lung lesions suggestive of malignancy. Although the determination of serum tumor marker levels is less accurate than FDG DHTC, the combination of a positive FDG result and positive tumor markers may be helpful in improving the diagnosis of lung cancer. PMID- 12775262 TI - [The effect of therapeutic intervention with corticosteroids on outcome and prognosis of sarcoidosis]. AB - OBJECTIVE: To evaluate the effects of corticosteroids on the prognosis of sarcoidosis. METHOD: There were analysis and comparison on the prognosis of natural course and corticosteroids therapy of 133 patients with sarcoidosis diagnosed by biopsy pathologic histology follow-up over 4 years. RESULTS: Six of the 7 patients with ocular sarcoidosis were cured by oral prednisone (PRED), and 1 patient receiving intraocular dexamethasone lost his sight. In patients with extra-pulmonary sarcoidosis who were not treated, 33 of the 38 cases underwent spontaneous remission, but another 5 cases progressed. Twenty-two of the 26 patients with stage I pulmonary disease underwent spontaneous remission, 3 progressed, with only one case cured by PRED therapy. In the 55 patients with stage II pulmonary sarcoidosis who received PRED therapy, the chest roentgenogram and lung function were back to normal in 45 cases, but 10 cases failed. One patient with stage II pulmonary disease without treatment progressed to stage IV. Three patients with stage III pulmonary sarcoidosis were cured by PRED therapy, but III patients with stage IV sarcoidosis failed to respond to PRED therapy. CONCLUSIONS: Systematic therapy with oral corticosteroids for ocular and stage II approximately III pulmonary sarcoidosis is a curative intervention. Corticosteroids can relieve symptoms, reduce inflammation, and improve the prognosis of ocular and pulmonary sarcoidosis. PMID- 12775263 TI - [The role of delayed rectifier potassium channels in the regulation of bronchial smooth muscle tension in asthmatic rats]. AB - OBJECTIVE: To investigate the role of delayed rectifier potassium channels (K(V)) in the regulation of bronchial smooth muscle tension in asthmatic rats and their effects on bronchial reactivity in vitro. METHODS: By using a specific potassium channel blocker, the bronchial contraction induced by spasmogens was observed in normal and asthmatic rats with the isometric tension recording technique. RESULTS: (1) K(V) blocker 4-aminopyridine (4-AP) caused concentration-dependent bronchial contraction in vitro. The concentration-response curve to 4-AP in the asthmatic group showed a left displacement with bigger pD(2) (the negative logarithm of the drug concentration causing 50% of maximal effect, 2.58 +/- 0.07, n = 10, P < 0.001) and no change in E(max) [maximal effect, (32 +/- 5) mg/mg, P > 0.05], compared with pD(2) (2.12 +/- 0.04, n = 10) and E(max) [(31 +/- 6) mg/mg] obtained in the control group; (2) In the control group, pre-treatment with 0.1 mmol/L 4-AP displaced the concentration-response curves to ET-1 and histamine to the left. pD(2) (for ET-1 and histamine) before and after 4-AP treatment was 6.27 +/- 0.38, 5.59 +/- 0.27 and 6.80 +/- 0.47, 6.42 +/- 0.14 respectively (P < 0.01). E(max) before and after 4-AP treatment was (36 +/- 8) mg/mg, (36 +/- 8) mg/mg and (40 +/- 8) mg/mg, (39 +/- 9) mg/mg respectively (P > 0.05); (3) In the asthmatic group, pre-treatment with 0.1 mmol/L 4-AP had no effect on the concentration response curve to ET-1 and histamine. There was no significant difference (P > 0.05) in pD(2) before (6.51 +/- 0.07, 5.86 +/- 0.14 respectively) and after (6.48 +/- 0.16, 5.96 +/- 0.08 respectively) 4-AP treatment, and in E(max) before [(61 +/- 8) mg/mg, (54 +/- 11) mg/mg respectively] and after [(65 +/- 10) mg/mg, (55 +/- 9) mg/mg respectively] 4-AP treatment. CONCLUSION: The activity of K(V) decreases in bronchial smooth muscle isolated from asthmatic rats compared with that obtained from normal ones. This change may be involved in the bronchial hyperreactivity to some spasmogens in vitro in asthmatic rats. PMID- 12775264 TI - [The study of inflammatory mediators and pulmonary ventilatory capacity in patients with chronic obstructive pulmonary disease]. AB - OBJECTIVE: To study the correlation between the inflammatory mediators released by alveolar macrophages and the pulmonary ventilatory capacity in patients with chronic obstructive pulmonary disease (COPD). METHODS: Alveolar macrophages were collected by fiberoptic bronchoscopy from 8 patients with chronic bronchitis, 8 with COPD with a forced expiratory volume in one second (FEV(1)) < or = 70%, and 8 healthy nonsmokers. All patients were in the stable stage. The macrophages were cultured and stimulated with lipopolysaccharide (LPS, 10 microg/ml). IL-8, IL-1 beta, TNF-alpha and IL-6 in the supernatants were measured by ELISA. Pulmonary functions were tested in all three groups. The correlation between different cytokines was tested with Pearson's relevant analysis, and the correlation between lung functions and cytokines was tested with multiple reverse regression analysis. RESULTS: (1) The concentrations of IL-8 released from macrophages in the COPD group were (43 +/- 27) microg/L and (57 +/- 41) microg/L (with LPS), higher than those from healthy controls [(13 +/- 10) microg/L and (20 +/- 13) microg/L] (P < 0.05), but not different from those in patients with chronic bronchitis [(29 +/- 21) microg/L and (32 +/- 23) microg/L] (P > 0.05). (2) The concentrations of IL-1 beta released from macrophages in the COPD group, the chronic bronchitis group and the control group were [(50 +/- 41) ng/L, (94 +/- 59) ng/L, (37 +/- 32) ng/L] before LPS, and [(225 +/- 108) ng/L, (153 +/- 175) ng/L, (70 +/- 37) ng/L] after LPS stimulation, which were positively correlated with the concentrations of IL-8 (P < 0.05). The concentrations of TNF-alpha released from macrophages in three groups were [(1,238 +/- 679) ng/L, (3,088 +/- 2,879) ng/L and (1,332 +/- 1,846)ng/L], which were positively correlated with the concentrations of IL-1 beta (P < 0.05). The concentrations of IL-6 released from macrophages in the three groups were [(7,959 +/- 8,458) ng/L, (5,317 +/- 10,112) ng/L and (6,480 +/- 4,982) ng/L, which were positively correlated with the concentrations of IL-8 (P < 0.05). (3) The values of FEV(1)/FVC, V(max50) and V(max25) measured in the COPD group were [(65.1 +/- 5.3)%, (43 +/- 8)% and (37 +/ 11)%, respectively, which were negatively correlated with the concentrations of IL-8 (P < 0.05), but negatively correlated with the concentrations of IL-1 beta only in the presence of LPS (P < 0.05). CONCLUSION: IL-8 released by alveolar macrophages plays an important role in the process from chronic cough to chronic airflow obstruction. TNF-alpha, IL-1 beta and IL-6 released by alveolar macrophages are also involved in the airway inflammation in COPD. PMID- 12775265 TI - [Effect of aquaporin-1 and sodium channel on pleural fluid transport in mice]. AB - OBJECTIVE: To investigate the effect of sodium channel and aquaporin-1 on pleural fluid transport. METHODS: Mice were briefly anesthetized, and the pleural space was infused with 0.25 ml of hyperosmolar or isosmolar solution, with terbutaline or amiloride in instilled solution. The mice were treated with either intraperitoneal dexamethasone or intravenous HgCl(2). Mice were euthanized at specified times for measurement of pleural fluid osmolality, volume and (125)I album in concentration. The rate of pleural fluid filtration and clearance was calculated, which was used to indicate the effect of sodium channel regulator (amiloride and terbutaline) and aquaporin regulator (dexamethasone and HgCl(2)) on pleural fluid transport. RESULTS: Amiloride decreased the osmotically driven water transport and isosmolar fluid clearance in pleural space (P < 0.05). Terbutaline increased osmotically driven water transport and isosmolar fluid clearance in pleural space (P < 0.05). HgCl(2) decreased, while dexamethasone increased, the osmotically driven water transport in pleural space (P < 0.05). HgCl(2) and dexamethasone had no effect on pleural isosmolar fluid clearance (P > 0.05). CONCLUSIONS: Sodium channel may play a role in osmotic and isosmolar fluid transport in pleural space. Aquaporin-1 may play a role in osmotic fluid transport in pleural space, while having no effect on isosmolar fluid clearance. PMID- 12775266 TI - [Rearrangement and altered expression of actin in macrophages induced by Mycobacterium tuberculosis]. AB - OBJECTIVE: To investigate the effect of M. tuberculosis infection on actin in host-cells. METHODS: The form and distribution of fibrous actin and changes of actin expression were observed by confocal microscopy and Western blot analysis in macrophages infected with either M. tuberculosis H(37)R(a) or H(37)R(v) at 6 h, 12 h and 24 h after infection. Non-infected macrophages or macrophages treated with dead M. tuberculosis H(37)R(v) served as controls. RESULTS: F-actin aggregated and the actin expression was suppressed in macrophages infected with H(37)R(a) or H(37)R(v), and the effect appeared earlier in cells infected by the virulent H(37)R(v) strain than in cells infected by the avirulent H(37)R(a) strain. In macrophages treated with the dead H(37)R(v), actin was not affected. CONCLUSION: The results suggest that in the process of infection, M. tuberculosis evades the bactericidal mechanisms possibly by secretion of certain proteins or factors which affect the host-cell actin. PMID- 12775267 TI - The Patient Distress Scale questionnaire: factor structure and internal consistency. AB - PRIMARY OBJECTIVE: This study examined the factor structure and internal consistency of the Patient Distress Scale (PDS). The PDS is an 11-item questionnaire that was developed to assess acute neurological patients' awareness of emotional disturbances during early rehabilitation. RESEARCH DESIGN: A retrospective cohort of patients seen for inpatient rehabilitation following brain injury. PROCEDURES: The PDS was administered to patients and family members upon admission to the neurorehabilitation unit as part of their neuropsychological evaluation. OUTCOMES AND RESULTS: Subjects were heterogeneous sample of acute neurological inpatients (n = 84) and their relatives (n = 64). A principle components factor analysis with varimax rotation yielded a three factor solution, which together accounted for 62% of the variance. The factor solution provided partial support for the initial structure of the PDS. Analyses revealed relatively strong reliability coefficients for patients (r = 0.82) and relatives (r = 0.86) on the PDS total score. Acceptable reliability coefficients were also found for patients and relatives on each of the factor scores. CONCLUSIONS: Preliminary findings support the utility of the PDS as a measure of awareness in acute neurological patients. PMID- 12775268 TI - The relationship between pre-hospital and emergency department Glasgow coma scale scores. AB - INTRODUCTION: Pre-hospital GCS scores are used to make critical patient care decisions and to fill in gaps in hospital-based TBI surveillance, but they may not be accurate. OBJECTIVE: To determine the relationship between pre-hospital (EMS-GCS) and emergency physician GCS scores (ED-GCS). METHODS: Prospective observational study of 60 TBI patients with a field GCS of 8-13 and age > 18. ED GCS, EMS-GCS, time of GCS and vitals signs were recorded. ANALYSIS: Simple and multiple linear regression. RESULTS: The median EMS-GCS was 13 and that for ED GCS was 15. There was a significant linear relationship between ED-GCS and EMS GCS (r = 0.45, p = 0.003). There was improvement in the prediction of ED-GCS when alcohol/drug use and age (but not time) were added to EMS-GCS. CONCLUSION: EMS GCS is usually two points lower than ED-GCS, but the correlation between them is strong and independent of the time between score determinations. These results could prevent unnecessary procedures based on the EMS-GCS and improve the accuracy of TBI surveillance. PMID- 12775269 TI - Who knows best? Awareness of divided attention difficulty in a neurological rehabilitation setting. AB - OBJECTIVE: To explore whether patients relearning to walk after acquired brain injury and showing cognitive-motor interference were aware of divided attention difficulty; whether their perceptions concurred with those of treating staff. DESIGN: Patients and neurophysiotherapists (from rehabilitation and disabled wards) completed questionnaires. Factor analyses were applied to responses. Correlations between responses, clinical measures and experimental decrements were examined. RESULTS: Patient/staff responses showed some agreement; staff reported higher levels of perceived difficulty; responses conformed to two factors. One factor (staff/patients alike) reflected expectations about functional/motor status and did not correlate with decrements. The other factor (patients) correlated significantly with dual-task motor decrement, suggesting some genuine awareness of difficulty (cognitive performance prioritized over motor control). The other factor (staff) correlated significantly with cognitive decrement (gait prioritized over sustained attention). CONCLUSIONS: Despite some inaccurate estimation of susceptibility; patients and staff do exhibit awareness of divided attention difficulty, but with a limited degree of concurrence. In fact, our results suggest that patients and staff may be sensitive to different aspects of the deficit. Rather than 'Who knows best?', it is a question of 'Who knows what?' PMID- 12775270 TI - Evaluating clients' perceptions of the quality of head injury rehabilitation services: development and validation of a questionnaire. AB - OBJECTIVE: The objective of this research was to develop a tool to assess clients' perception of the quality of head injury rehabilitation services. METHODS AND RESULTS: A questionnaire PQRS-Montreal 1.0 was developed in Montreal, Quebec using a standardized methodology that incorporated a new technique to reach consensus regarding the tool's contents. Five dimensions of care (i.e. ecological approach, client-centred approach, accessibility, quality of the service providers and continuity) and their corresponding standards were identified as pertinent to include in the tool and were subsequently validated by experts. A phase of item generation followed that led to the development of an 87 item prototype of the questionnaire. The questionnaire is to be administered during an interview and employs a 5-point rating scale. DISCUSSION AND CONCLUSION: The implications for the use of PQRS-Montreal 1.0 in the accreditation process of head injury rehabilitation services are discussed, as well as plans for future quantitative research in the development of the tool. PMID- 12775271 TI - Problem-solving by traumatically brain injured and neurologically intact subjects on an adaptation of the twenty questions test. AB - PRIMARY OBJECTIVE: RAPS (Rapid Assessment of Problem-Solving) is a clinical measure for assessing verbal problem-solving in hard-to-test patients or those that may not be able to tolerate a longer, more detailed assessment. The design of the test is based on Mosher and Hornsby's Twenty Question test, but RAPS contains several modifications to facilitate its use with brain-injured individuals. This study used RAPS to compare the verbal problem-solving ability of subjects that were neurologically intact and subjects that had chronic traumatic brain injuries. METHODS AND PROCEDURES: Twenty-one adults that were neurologically intact (NI) and 21 adults that had incurred a traumatic brain injury (TBI) matched for age, gender and education took part in the study. Before being tested with RAPS, participants signed an IRB-approved consent form and completed a battery of neurocognitive measures. RAPS entailed the solving of three verbal problems. Each problem involved an array of 32 pictures of common objects (e.g. football) arranged in a 4x 8 grid. The subjects were instructed to ask yes/no questions to determine which picture the examiner was 'thinking of '. Three scores were computed for each problem solved: number of questions asked, percentage of constraint-seeking questions, and question-asking efficiency scores for the first four questions. OUTCOMES: No learning effects across the problems were found for any of the RAPS measures. Scores were averaged across the three problems to determine group effects. Groups of TBI and NI subjects did not differ significantly in the number of questions asked in solving RAPS problems. Members of the NI group asked significantly more constraint-seeking questions (e.g. Is it an animal?) than those in the TBI group, and the subjects that had incurred brain injuries did more guessing than the NI group. Over 70% of the time, guessing took place after the semantic category containing the target picture was known to the subject. Guesses took the form of pseudo-constraint questions (e.g. Is it the animal with a long neck?) rather than frank guesses (e.g. Is it the giraffe?). These trends were seen for both groups. Question-asking efficiency scores, computed for the first four questions of each problem, reflected the amount of information gained by the subjects' questions. It was anticipated that subjects' questioning strategies would target larger rather than smaller number of pictures and systematically reduce the number of total pictures under consideration. Question-asking efficiency scores were significantly higher for the group of NI subjects. Both groups increased question-asking efficiency scores across the first four questions, and there was no significant group x question interaction. Further analysis of the question-asking efficiency scores revealed that questions from the group of NI subjects tended to target multiple categories of pictures and larger single semantic categories of pictures on the 32-item problem-solving board, whereas those from the group of TBI subjects often targeted smaller categories or portions of categories. CONCLUSIONS: Two meta-cognitive functions, planning and strategy shifting, appeared to explain most of the differences in the verbal problem-solving performance between the groups. Both groups, however, demonstrated a range of abilities on RAPS. Until a larger normative database for RAPS is available, it behooves clinicians using the test to analyse results on an individual basis, to consider the subject's pre-morbid problem-solving ability and to weigh those factors associated with brain injury that could affect RAPS performance. PMID- 12775272 TI - A pilot study exploring electronic (or e-mail) mail in users with acquired cognitive-linguistic impairments. AB - This paper reports the results of an exploratory study into the usability of a simplified e-mail interface for eight individuals with acquired cognitive linguistic impairments. Participatory Action Research, a qualitative research method, was used to capture the range of performance variables and to emphasize a 'user-centred' approach to the research process. The participants were asked to read and reply to e-mails across four writing prompt conditions. An analysis of errors and participant preferences for the prototype e-mail system was conducted. Errors fell into two general categories: (1) computer usability (e.g. conceptual understanding of mouse/cursor operation) and (2) message composition (e.g. generating ideas for a message). Participant preferences for the writing prompt conditions varied considerably. All participants endorsed the use of customized e mail interfaces as a means of connecting with friends and family, thereby reducing social isolation. Implications for interface design, rehabilitation and future research into assistive technology are discussed. PMID- 12775273 TI - Early recognition of neuroleptic malignant syndrome during traumatic brain injury rehabilitation. AB - Neuroleptic malignant syndrome is a rare disorder that manifests with hyperthermia, muscle rigidity and autonomic instability. Presented is a case series of individuals with traumatic brain injury and agitation who, when treated with neuroleptics, developed neuroleptic malignant syndrome. Although the incidence of this syndrome is rare, it is associated with significant morbidity and mortality. The onset of symptoms inconsistent with the patient's current level of recovery should alert the clinician to consider other possible diagnosis and failure to distinguish the features of neuroleptic malignant syndrome from post-traumatic agitation will delay appropriate intervention for this potentially life-threatening disorder. PMID- 12775274 TI - Meta-analyses of results of alarm treatment for nocturnal enuresis--reporting practice, criteria and frequency of bedwetting. AB - OBJECTIVE: The purpose of this article is to obtain a realistic picture of the efficiency of alarm treatment for nocturnal enuresis by means of meta-analysis (this is done by estimating a model for the success rate of the treatment as reported in the literature)and to interpret the results of the individual reports in the light of the model. MATERIAL AND METHODS: A model including year of treatment, applied criteria for success and the frequency of wet nights is proposed and calculated. RESULTS: The apparent decline in success over the period 1938-96 is explained in terms of systematic changes in reporting practice and underestimation of the problem of relapse. It is shown that the probability of the success of alarm treatment for nocturnal enuresis increases with the frequency of wet nights, confirming a previous finding of the authors. CONCLUSION: Meta-analyses can give a much more realistic picture of the efficiency of alarm treatment for nocturnal enuresis than can be obtained from an individual report. PMID- 12775275 TI - Telomerase activity in diagnosis of bladder cancer. AB - OBJECTIVE: Telomerase is an enzyme that can reconstitute the ends of chromosomes after cell division and thus circumvent the damage that occurs in normal adult somatic cells during successive mitotic cycles. Immortal cells have short but stable chromosomes and increased telomerase activity. Transitional cell carcinoma (TCC) has only a few useful markers of diagnostic or prognostic importance. The objectives of this study were to determine whether there was a correlation between telomerase activities and the grade or stage of TCC and whether the activity of the enzyme could serve as a biochemical marker of this tumor. MATERIAL AND METHODS: Telomerase activity was determined by examining, using a polymerase chain reaction-based assay designed using the telomeric repeat amplification protocol (TRAP), urine cell pellets obtained from 42 bladder cancer patients, 18 patients with primary hematuria, 19 patients with benign urologic disease, 14 patients with urologic malignancies other than TCC and 20 healthy volunteers. RESULTS: Telomerase activity was found in 24/31 patients with bladder tumors (77.4% sensitivity) and in 5/77 patients without tumors (93.5% specificity). No correlation was found between telomerase activity and the grade or stage of the tumor. Although none of the urine cell pellets obtained from the 20 healthy volunteers demonstrated telomerase activity, positive telomerase activity was found in two subjects in the benign urologic disease group and in three subjects in the other urologic malignancy group. It was demonstrated that gross hematuria was the cause of false-negative results in six of the nine patients (66.7%). but washing the pellets four times and diluting them before the TRAP assay solved this problem. CONCLUSION: These results indicate that telomerase activity may be a promising marker for TCC but the technical aspects of the technique must be improved before it is used in routine clinical practice as a standard method. False-negative results obtained using gross hematuric urine should be carefully reevaluated and cell pellets should be washed again and diluted before analysis. PMID- 12775276 TI - A population-based study of 538 patients with newly detected urinary bladder neoplasms followed during 5 years. AB - OBJECTIVE: To describe in detail the diagnosis and clinical course of an unselected population-based cohort of patients with newly diagnosed bladder neoplasms. MATERIAL AND METHODS: A total of 538 patients registered in the Stockholm region with newly diagnosed primary bladder neoplasms (transitional cell carcinomas) in 1995 and 1996 were followed for at least 5 years. All hospitals and urology units in the region participated in the study. Treatment and follow-up were performed according to a standard-of-care programme. Routine pathological reports were used. Original case records were scrutinized on location in 2001. In addition, a tumour bank of freshly frozen tumour tissue was established. RESULTS: The calculated 5-year cancer-specific survival rate for the 538 patients in the cohort was 78%. No patient (0/29) with TaG1 tumours showed progression or died of bladder cancer. Only 2/187 patients (1%) with stage Ta and grade 2A or 2B tumours died of bladder cancer. In contrast, after 5 years of follow-up, patients with TaG3 and T1G2B tumours had disease-specific death rates of 20% and 27%, respectively. The result of the first cystoscopy examination after the initial resection of non-invasive tumours was of prognostic value. Recurrent disease was present in 62% (248/402) of all patients with Ta and T1 tumours at diagnosis and patients with T1 tumours had recurrences earlier than those with Ta tumours. Moreover, 32% (35/110) of the patients who presented with T1 tumours at diagnosis progressed to muscle-invasive disease during the follow up period. The overall prognosis for patients presenting with muscle-invasive tumours (T2+) was dismal, with 69% (80/116) of the patients dying of the disease. CONCLUSIONS: We analysed a population-based cohort of patients with urinary bladder neoplasms in order to establish a clearly defined and unselected clinical series, with the main aims of comparing and evaluating the clinical utility of new molecular biology techniques. In the present series, TaG1 tumours behaved benignly. The disease-specific mortality rate was low for initial TaG2 tumours, intermediate for initial TaG3 and T1 tumours and high for initial T2+ tumours. PMID- 12775277 TI - Histopathologic changes in the mucosa of ileal orthotopic neobladder--findings in 24 patients followed up for 5 years. AB - OBJECTIVE: To evaluate changes occurring in the mucosa of the neobladder over a period of 60 months in 24 patients with orthotopic ileal neobladder. MATERIALS AND METHODS: A total of 36 male patients have undergone radical cystoprostatectomy and received an orthotopic ileal neobladder in our hospital during the last 10 years; 24 of these patients, all of whom completed a follow-up period of 60 months, are included in this study. All cases underwent a biopsy of the ileal mucosa at the time of surgery and their neobladder mucosa was then biopsied at 6, 12, 24, 36 and 60 months. The specimens were stained with hematoxylin-eosin in order to examine the changes in the thickness of the mucosa and its villi. Sections were also stained with Alcian blue and periodic acid Schiff in order to determine the number of goblet cells. A morphometric scoring system was created to quantify the change in villi size. RESULTS: The mean thickness of the ileal mucosa at the initial biopsy was 270.9 +/- 35.9 microm and displayed a continuous decrease at all of the subsequent biopsies. The number of luminal goblet cells increased during follow-up. Villus atrophy was found to be a continuous process during follow-up. No dysplasia or malignancy was detected in any of the biopsies. CONCLUSION: During a follow-up period of 5 years, there were no neoplastic changes in our patients, but instead a protective response of the mucosa to its new environment was observed. PMID- 12775278 TI - Population-based study of prostate-specific antigen testing and prostate cancer detection in clinical practice in northern Sweden. AB - OBJECTIVE: The pattern of prostate-specific antigen (PSA) testing in clinical practice is largely unknown; it may be used either in asymptomatic men or in the work-up of men with urinary or other symptoms. The aim of this study was to investigate the pattern of PSA testing in clinical practice for men diagnosed with stage T1c prostate cancer in a region with no formal screening programme. MATERIAL AND METHODS: Using the Primary Prostate Cancer Register for Northern Sweden, all cases of stage T1c prostate cancer diagnosed between 1992 and 1999 in the city of Umea were identified. The cause of PSA testing was assessed by examining the medical records. Men were categorized as asymptomatic, having lower urinary tract symptoms (LUTS) or having symptoms other than LUTS. Prospective registration of the cause of PSA testing in the entire region of Northern Sweden started in 2000. RESULTS: We found that in Umea only 32/213 (15%) cases diagnosed with T1c prostate cancer were asymptomatic at the time of PSA testing, 55% of men had a PSA test as part of a work-up for LUTS and 30% had other symptoms. In 2000, 126 cases of stage T1c prostate cancer were diagnosed in the entire region and 20/126 (16%) of these men were asymptomatic. CONCLUSIONS: PSA testing was mostly used as a tool in the work-up of symptomatic patients in Umea and also in the region of Northern Sweden. Further studies in other populations are needed. PMID- 12775279 TI - Does initial surveillance in early prostate cancer reduce the chance of cure by radical prostatectomy?--A case control study. AB - OBJECTIVE: To evaluate whether initial surveillance followed by prostatectomy impairs pathological stage compared to immediate surgery in men with prostate cancer detected as a result of early screening. MATERIAL AND METHODS: A total of 26 patients with prostate cancer [T1c-T2, Gleason score <7, prostate-specific antigen (PSA) 3-13 ng/ml] who were managed by means of initial surveillance (mean 23.4 months, range 8-55 months) followed by radical retropubic prostatectomy (RRP) were evaluated. For each of these cases two matched control cases were selected from patients who were operated on without prior surveillance. The two groups were matched for PSA, age, T stage and Gleason score at biopsy. Evaluation of prostatectomy specimens included measurement of tumour volume, pT stage and Gleason score. RESULTS: Tumour volume did not differ significantly between cases and controls: 1.35 vs 1.05 cm (3), respectively. The frequency of extracapsular growth, Gleason score and time to progression after RRP within a mean follow-up period of 2 years were also similar between the two groups. CONCLUSION: In selected patients with very early prostate cancer it seems that close surveillance followed by prostatectomy when signs of progression appear is a low risk option. However, before this strategy can be generally recommended longer follow-up periods should be used and a randomized study should be performed. PMID- 12775280 TI - Frequency of lymphoceles after open and laparoscopic pelvic lymph node dissection in patients with prostate cancer. AB - OBJECTIVE: To compare the frequencies of pelvic lymphocele formation after laparoscopic and open pelvic lymph node dissection in patients with prostate cancer. MATERIAL AND METHODS: A total of 132 patients operated on with pelvic lymph node dissection (PLND) underwent CT scanning of the abdomen and pelvis at a median of 29 days postoperatively. Open pelvic lymph node dissection (OPLND) was performed in 94 patients (71%) and 38 patients (29%) were operated on using a laparoscopic technique (LPLND). The frequency and size of pelvic lymphoceles were registered. Lymphoceles with a horizontal diameter of /=5.0 cm were classified as large. RESULTS: The overall frequency of lymphoceles was 54%. The frequencies in the OPLND and LPLND groups were 61% and 37%, respectively. A total of 27% of the OPLND patients had large lymphoceles, compared to 8% of the LPLND patients. Three patients (2.3%), all in the OPLND group, had clinically significant lymphoceles. CONCLUSIONS: Although the overall frequency of lymphocele formation was high, clinically significant lymphoceles were scarce. LPLND was associated with a statistically significant lower frequency of lymphocele formation compared to OPLND. PMID- 12775281 TI - MRI of the skeleton in prostate cancer staging. AB - OBJECTIVE: To explore the value of MRI in the detection of bone metastases in newly diagnosed prostate cancer. MATERIAL AND METHODS: MRI examinations of the axial skeleton in 76 patients with newly diagnosed prostate cancer were reviewed, and the relation of these findings to the serum level of prostate specific antigen (PSA) was examined. RESULTS: MRI indicated bone metastases in 26/76 patients (34%) in the entire study group, in 4/24 (17%) with serum PSA <20 ng/ml and in 22/52 (42%) with serum PSA >20 ng/ml. CONCLUSIONS: These results suggest that MRI is a more sensitive indicator of suspected bone metastases than bone scintigraphy in the low range of serum PSA, but less sensitive in the high range. Further studies of MRI and bone scintigraphy in parallel in patients with serum PSA <20 ng/ml are needed to elucidate their relative value in the staging of patients with prostate cancer. PMID- 12775282 TI - Technological changes in the management of prostate cancer result in increased healthcare costs--a retrospective study in a defined Swedish population. AB - OBJECTIVE: In two previous studies we calculated direct costs for men with prostate cancer who died in 1984-85 and 1992-93, respectively. We have now performed a third cost analysis to enable a longitudinal cost comparison. The aim was to calculate direct costs for the management of prostate cancer, describe the economic consequences of technological changes over time and estimate total direct costs for prostate cancer in Sweden. MATERIAL AND METHODS: A total of 204 men in a defined population with a diagnosis of prostate cancer and who died in 1997-98 were included. Data on utilization of health services were extracted from clinical records from time of diagnosis to death from a university hospital and from one county hospital in the county of Ostergotland. RESULTS: The average direct cost per patient has been nearly stable over time (1984-85: 143 000 SEK; 1992-93: 150 000 SEK; 1997-98: 146 000 SEK). The share of costs for drugs increased from 7% in 1992-93 to 17% in 1997-98. The total direct costs for prostate cancer in Sweden have increased over time (1994-85: 610 MSEK; 1992-93: 860 MSEK; 1997-98: 970 MSEK). CONCLUSIONS: Two-thirds of the total cost is incurred by inpatient care. The share of the total costs for drugs is increasing due to increased use of gonadotrophin-releasing hormone analogues. Small changes in average direct costs per patient despite greater use of technology are explained by the fact that more prostate cancers are detected at the early stages. PMID- 12775283 TI - Children presenting at UK community enuresis clinics--comparison with hospital based samples. AB - OBJECTIVE: This study compares the clinical and psychosocial characteristics and the treatment outcomes of bedwetting UK children presenting at a community enuresis service with those from studies conducted in hospital-based settings. MATERIAL AND METHODS: Cluster stratification by clinic was applied to ensure that the population attending the 15 enuresis clinics selected was representative. Parents completed the maternal tolerance scale and children completed the impact of bedwetting and Coopersmith self-esteem scales. The electronic databases MEDLINE and CINAHL were searched for the years 1966-2002 for UK-based empirical studies conducted in children aged 5-16 years with nocturnal enuresis. RESULTS: Children in the community sample were younger and had more day-time wetting than the hospital-based population but did not have significantly lower self-esteem. The impact of bedwetting had the strongest relationship with the Coopersmith self esteem score, followed by ethnicity and the maternal tolerance score (beta = 0.49, p < 0.001; beta = 2.83, p < 0.001; and beta = 0.45, p = 0.015; respectively). CONCLUSION: Primary referrals to community enuresis services are younger and have more day-time wetting than those attending hospital-based clinics but the majority do not have low self-esteem. Ethnicity appears to be an important factor in evaluating the impact of wetting on the child. PMID- 12775284 TI - Comparison of five different hormonal treatment protocols for children with cryptorchidism. AB - OBJECTIVE: To compare the effects of medical therapy on boys with cryptorchidism, a prospective study was carried out in five groups of patients over a 30-month period. MATERIAL AND METHODS: A total of 439 boys with undescended testicles were enrolled; their ages ranged between 6 months and 13 years (median 4.2 years). Of these, 327 had true unilateral and 112 bilateral undescended testicles. In 115 patients surgery was performed directly either because the condition was associated with inguinal hernia or because the child had undergone previous inguinal surgery; hormonal treatment was attempted in the remaining 324 patients. Human chorionic gonadotrophin (hCG) was administered to 113 patients (34.8%), luteinizing hormone-releasing hormone (LH-RH) to 85 (26.2%), a combination of LH RH and hCG to 64 (19.7%), human menopausal gonadotrophin (hMG) to 35 (10.8%) and a combination of hMG and hCG to 27 (8.3%). RESULTS: Overall, testicular descent was achieved in 27.7% (90/324) of patients: specifically, in 38.2% (36/94) of boys with bilateral undescended testicles (both testes in 22/36, one testis only in 14/36) but in only 23.4% (54/230) of those with unilateral undescended testicles (p = 0.007). The correlation between the type of therapy and testicular descent can be summarized as follows: hCG, 39/113 (34.5%); LH-RH, 25/85 (29.4%); hCG+LH-RH, 19/64 (29.6%); hCG+hMG, 7/27 (25.9%); hMG alone, 0/35 (0%). CONCLUSIONS: In our experience, pharmacological treatment seems to yield better results in cases of bilateral than unilateral cryptorchidism. hCG seems to be equally effective as LH-RH for the treatment of cryptorchidism in approximately 30% of cases; the combination of these two drugs does not seems to increase the success rate. The use of hMG alone is ineffective. We believe that hormonal treatment can lead to acceptable results in boys with cryptorchidism, without relevant adverse effects. PMID- 12775285 TI - Retrospective analysis of inflammatory parameters in acute pyelonephritis. AB - OBJECTIVE: Different parameters were compared in children with acute pyelonephritis in order to facilitate the early detection of vesicoureteral reflux (VUR) in primary healthcare settings. MATERIAL AND METHODS: This was a cross-sectional, hospital-based study. A total of 149 children with confirmed acute pyelonephritis were retrospectively analyzed between January 1999 and December 2000. The primary factors studied were body temperature, C-reactive protein level, white blood cell count and neutrophil ratio. RESULTS: The study population comprised 82 males (55.0%) and 67 females (45.0%). The mean age of the patients was 2.7 +/- 3.9 years (age range 1 day to 18 years; 75% <3 years old). Of 123 patients who underwent voiding cystourethrography, 34 (27.6%) had VUR. When raised C-reactive protein, leukocytosis and raised neutrophil ratio occurred together, the specificity and positive predictive value for predicting VUR were obviously increased, but sensitivity was radically decreased. After controlling for the other covariates, multivariate logistic regression analysis showed that factors significantly related to VUR were age (odds ratio = 1.3, 95% confidence interval 1.02-1.67; p < 0.05) and raised neutrophil ratio (odds ratio = 4.2, 95% confidence interval 1.1-16.5; p < 0.05). CONCLUSIONS: Our findings emphasize that the prevalence of VUR in children with acute pyelonephritis is extremely high. If a raised neutrophil ratio is observed, the potential risk of VUR is significantly increased in patients with clinically suspected acute pyelonephritis. Hence, the neutrophil ratio is recommended as an excellent parameter for predicting VUR. PMID- 12775286 TI - Interstitial cystitis: a review of current concepts of aetiology, diagnosis and therapy. AB - Until recently, interstitial cystitis (IC) has been described as a chronic clinical syndrome of obscure aetiology and, consequently, specific treatment for the condition is lacking. IC has often been regarded as a diagnosis of exclusion in women complaining of chronic pelvic pains, frequency, dysuria, etc. and until recently most clinicians rarely considered this diagnosis in men. However, recent research efforts have begun to yield vital information on this underdiagnosed and distressing clinical condition regarding its aetiology and diagnosis and, consequently, more effective therapeutic options are now emerging. The aim of this review is to highlight these emerging concepts of the aetiology, diagnosis and innovative therapy of IC, in order to help clinicians confronted by patients with this disorder. PMID- 12775287 TI - Presence and significance of TT virus in Danish patients on maintenance hemodialysis. AB - OBJECTIVES: To determine the prevalence of TT virus (TTV) in a population of Danish hemodialysis patients and evaluate possible relations between TTV infection and elevated levels of C-reactive protein (CRP) and hypo-response to treatment with erythropoietin (EPO). MATERIAL AND METHODS: Patients on maintenance hemodialysis at a single center were invited to participate. Demographic and clinical data were registered. Blood samples for virological and routine biochemical tests were drawn simultaneously. TTV DNA was detected using polymerase chain reaction (PCR). TTV viral load was estimated by means of semi quantitative PCR. All patients were tested for hepatitis B, hepatitis C and GB virus C. RESULTS: Of 252 patients, 204 (80.9%) gave their written informed consent to participate in the study. The prevalence of TTV was 68% and 50% of TTV positive patients had a high TTV viral load. TTV-positive patients were significantly older than TTV-negative patients (p = 0.011). No relations were found between TTV infection and elevated levels of alanine aminotransferase (ALT) or CRP or hypo-response to EPO treatment. The mean hemoglobin concentration was 11.24 +/- 1.48 g/dl. Patients with a high TTV viral load had a lower level of hemoglobin (10.86 +/- 1.47 g/dl) than the others (p = 0.01). This trend suggested a positive relation between TTV infection and the number of blood transfusions. A restriction fragment length polymorphism assay suggested that patients were infected with different TTV strains. CONCLUSIONS: TTV is common in patients on maintenance hemodialysis. The presence of TTV is associated with increasing age. Patients with a high TTV viral load had lower levels of hemoglobin than the others. TTV infection is not related to elevated levels of ALT or CRP or to hypo response to EPO treatment. PMID- 12775289 TI - Squamous cell carcinoma and transitional cell carcinoma arising in bladder augmentations. AB - Tumours arising in bowel-augmented bladders are rare. Usually these tumours are adenocarcinomas that occur along the anastomotic line. We present two unusual tumours, squamous cell carcinoma and transitional cell carcinoma, that occurred in bladder augmentations. We also emphasize the need for regular cystoscopic surveillance. PMID- 12775288 TI - Hypercalcemia upon recurrence of renal cell carcinoma producing parathyroid hormone-related protein. AB - We established a new renal carcinoma cell line that produces parathyroid hormone related protein (PTHrP) and interleukin-6 in culture. The cellular production of PTHrP was confirmed by Northern blot analysis and immunofluorescence examination. Bone and lung metastases occurred simultaneously 3.5 years after surgery. The patient did not show hypercalcemia at this time, despite the presence of multiple osteolytic metastases. About 7 months after bone metastasis was first shown, serum PTHrP was detected by means of an immunoradiometric assay and the calcium level was found to be elevated to 3.29 mmol/l. The hypercalcemia was successfully controlled by i.v. administration of bisphosphonates. PMID- 12775291 TI - Child war trauma: a comparison of clinician, parent and child assessments. AB - This paper focuses on the difficulty of capturing child war trauma: the appropriateness of a standardized trauma questionnaire and the value of recruiting multiple reports. Three independent assessments of the war exposure of 75 Bosnian refugee children and teenage youths (aged 1-20), resettled in Sweden, are compared: clinician assessment based upon a semi-structured interview with the family, child self-report on the Harvard Trauma Questionnaire (HTQ) and parent report on the same questionnaire. Parent and clinician reports show marked group similarities but differ often with regard to the individual child. Clinician score reveals a social class gradient not visible on the HTQ. Parent and teenager assessments correlate strongly on total exposure but diverge markedly on specific events. Discrepancy derives as frequently from events affirmed by teenager alone as by parent alone. Primary school children, on the other hand, systematically offer a less-detailed account of their own war exposure. In summary, original HTQ functions "quite well" as a standardized questionnaire, but a Bosnian-specific version would expectedly afford greater validity and capture social class differences in child exposure. For teenagers, the value of multiple informants appears evident; for primary school children, a more adequately age-adjusted procedure remains the first priority. PMID- 12775292 TI - Screening for post-traumatic stress disorder among refugees in Stockholm. AB - A screening procedure (The Health Leaflet; HL) to assist social workers in finding subjects with possible post-traumatic stress disorder (PTSD) in recently resettled refugees is presented. It is compared with two established self-rating instruments, the Harvard Trauma Questionnaire (HTQ) and Impact of Event Scale-22 (IES-22), as well as structured clinical interview. AIM: To validate the screening interview and the rating scales in comparison to a clinical assessment for PTSD, and examine the feasibility of lay screening for PTSD. FINDINGS: The Health Screening Interview with a cut-off value of 10 points identified cases with fully developed PTSD with both sensitivity and specificity about 0.7. Only two items--difficulties concentrating and having been exposed to torture- contributed to the discriminatory performance of the HL interview. In the HTQ symptom subscale, emotional detachment and a feeling of going mad contributed to the discriminatory performance. In the IES-22, recurring strong affects about the events, as well as intrusive memories, were the items with the highest canonical correlation coefficients. In the HL, the single screening question about difficulties concentrating identified 31/32 individuals diagnosed with PTSD in this group, with a relative risk of 24. CONCLUSIONS: A mental health screening procedure during refugee reception performed by lay persons is clearly feasible and can assist in identifying subjects with trauma-related healthcare needs, thus leading to more realistic demands in refugee reception. PMID- 12775294 TI - Comorbid alcohol addiction increases aggression level in soldiers with combat related post-traumatic stress disorder. AB - The aim of this study was to compare aggressive behavior in soldiers with combat related post-traumatic stress disorder (PTSD), PTSD comorbid with alcohol addiction and alcohol addiction only. Three groups of male combat experienced soldiers with PTSD (n=43), PTSD comorbid with alcohol addiction (n=41) and alcohol addiction (n=39) were compared by Aggression rating scale A-87. PTSD was diagnosed according to DSM-IV criteria and Watson's PTSD rating scale. Alcohol addiction was diagnosed according to DSM-IV criteria and CAGE Questionnaire. Combat-experienced soldiers with alcohol addiction as well as soldiers with combat-related PTSD comorbid with alcohol addiction have a high level of verbal latent aggression (VLA), (F=26.65; P<0.001), physically latent aggression (PLA), (F=37.86; P<0.001), indirect aggression (INA), (F=56.94; P<0.001), verbal manifest aggression (VMA), (F=18.35; P<0.001), and physically manifest aggression (PMA), (F=43.22; P<0.001), vs. soldiers with combat-related PTSD without comorbid conditions. Alcohol addiction is a severe factor in increasing aggression levels in soldiers with PTSD. PMID- 12775293 TI - Cognitive abilities related to post-traumatic symptoms among refugees from the former Yugoslavia in psychiatric treatment. AB - The overall aim was to study the relationship between post-traumatic symptoms and cognitive abilities among traumatized refugees from the former Yugoslavia, in psychiatric treatment. The results showed that a diagnosis of post-traumatic stress disorder (PTSD), as well as a higher level of post-traumatic symptoms, was significantly associated with poorer average cognitive performance. Three of four tests of fluid intelligence, and the Benton Visual Retention Test, assessing episodic memory, were the most discriminating. A specific constellation of PTSD symptoms, dominated by arousal and intrusive symptoms, had a significant overall correlation with intellectual performance. One implication of the study is that assessment of cognitive abilities might be advisable in this patient group, in particular when arousal and re-experiencing symptoms are frequent. PMID- 12775295 TI - Refugee families during asylum seeking. AB - The mental health of refugee families with children during the asylum period is a neglected research area in psychiatry. The present paper describes the situation of 10 refugee families residing at an asylum centre in Finland. Case vignettes are presented to illustrate the situations of these families. The study shows a high rate of depressive and post-traumatic stress disorder-related symptoms among adult refugees. The case vignettes suggest that during the asylum period, many children and adult members of the family are not in a post-traumatic situation, but they live constantly in a distressing situation. The foremost distress amongst the asylum seekers appeared to be fear of deportation and separation from family members. Most of the adults and all children had not received any psychiatric or psychotherapeutic assessment or treatment. It is likely that current procedures for dealing with the asylum seekers contributes to the level of stress, family confusion and psychiatric problems in already traumatized refugee families. PMID- 12775296 TI - Stressful life events preceding the first onset of psychosis. An explorative study. AB - The aim of the study was to investigate the frequency and types of psychologically stressful life events preceding first episode psychoses. In a defined urban area, all first-episode psychosis patients aged 18-45, without signs of organic brain disorder or heavy abuse, were investigated and followed-up during 2-3 years. In the majority of cases, significant releasing factors could be depicted. The need to understand the specific, private meaning of the preceding events is illustrated by case vignettes. A stressful life situation may not be regarded as the "cause" of the psychosis but as a more or less important boosting factor. This is a complicated interaction between biological and psychological vulnerability factors. It seems important that the patients after the acute phase are given opportunities to talk about the events and how to manage them in the future. PMID- 12775297 TI - Ward atmosphere in acute psychiatric inpatient care: differences and similarities between patient and staff perceptions. AB - The purpose of the study was to compare patient and staff perceptions of the social climate in a psychiatric department, in order to investigate to what extent the two groups agree in their view of the ward milieu. Ninety-six patients and 66 staff members answered the Ward Atmosphere Scale (WAS) questionnaire, comprising a 100-item "real-form" concerning the ward atmosphere as it is actually perceived and a 100-item "ideal-form" concerning the atmosphere in an imagined "best of all" ward. Staff scores were significantly higher than patient scores on the WAS-R subscales Involvement, Spontaneity, Practical orientation, Personal problems orientation and Program clarity, and on the WAS-I subscales Spontaneity, Practical orientation, and Anger and aggression. Patient scores were significantly higher than staff scores on the WAS-R subscale Staff control and on the WAS-I subscales Support, Autonomy and Staff control. At the same time, the findings showed that the WAS-profiles of the two groups were highly concordant regarding both "realities" and "ideals". In conclusion, patients and staff do to some extent differ in their perception of the social climate in the ward. However, the differences seem to reflect nuances in views rather than fundamental differences in opinion. From an overall point of view, the findings suggest a basic agreement between patients and staff regarding the valuation of the treatment environment as well as the general preferences concerning the ward milieu. PMID- 12775298 TI - Dyslexia and psychosocial factors. A follow-up study of young Norwegian adults with a history of dyslexia in childhood. AB - The present study is a follow-up of two different groups of young adults with a history of dyslexia problems in childhood. Group A was drawn from a larger longitudinal study where students were diagnosed with dyslexia at age 10. Group B was recruited at a child psychiatric clinic and also had dyslexic problems. Measures of educational level, life satisfaction and psychosocial factors were applied. The assessment was carried out by means of tests, questionnaires and personal interviews. The results showed lower levels of educational attainment in group B as well as lower satisfaction with health, friends and education compared to group A and a normative group. Both dyslexic groups showed more psychiatric problems than those in the normative sample. PMID- 12775299 TI - Efficacy of repetitive transcranial magnetic stimulation in depression: a review of the evidence. AB - Repetitive transcranial magnetic stimulation (rTMS) is a novel treatment in psychiatry. We reviewed all published evidence on the efficacy of this treatment option in depressive disorders. An extensive electronic and manual search for eligible research reports identified only 12 studies that met the predetermined criteria for inclusion. rTMS was administered differently in most studies, and patient characteristics varied widely. A formal meta-analysis of the studies was thus not possible. Instead, we conducted a qualitative evaluation of the included studies. The antidepressive efficacy was not consistent, and where efficacy was demonstrated, it was modest in most studies. Some patients had good but transient responses to rTMS. Treatment gains were not maintained beyond the treatment period. Comparisons with electroconvulsive therapy (ECT) indicated the superiority of ECT. More, larger and more carefully designed studies are needed to demonstrate convincingly a clinically relevant effect of rTMS. We conclude that there is insufficient evidence for rTMS as a valid treatment for depression at present. PMID- 12775300 TI - Comparison of the GHQ-36, the GHQ-12 and the SCL-90 as psychiatric screening instruments in the Finnish population. AB - The aim of the study was to compare the screening properties of two General Health Questionnaire (GHQ) versions and the Symptom Checklist (SCL-90), and to evaluate them as psychiatric screening instruments in Finland. We administered the GHQ-36 and the SCL-90 to psychiatric outpatients (n=207) and to a community sample (n=315). Receiver operating characteristic (ROC) analysis was used to estimate the screening performance of the two instruments and of the GHQ-12 extracted from the GHQ-36. The screening properties of the scales were found to be good and similar. Suggested optimal cut-off points were 3/4 for the GHQ-12, 8/9 for the GHQ-36 and 0.90/0.91 for the SCL-90. In conclusion, the scales functioned equally well in screening. This favors the GHQ-12 for pure screening. When information on the symptom level is also needed, the GHQ-36 and the SCL-90 become better choices. The cut-off points presented here should be considered in the future Finnish psychiatric screening studies. PMID- 12775302 TI - The general practitioner and the preferable antihypertensive agent. PMID- 12775303 TI - Long-axis motion and left ventricular function. PMID- 12775305 TI - Clinical efficacies of antihypertensive drugs. AB - OBJECTIVE: According to published data, the ability to prevent various hypertension-related events differs between the various antihypertensive drug groups. Although absolute drug effects differ among studies, relative drug effects could be considered constant. We therefore explored the possibility of drawing statistically valid conclusions about the differences in clinical efficacy between various drug groups by doing an overview of published data. DESIGN: We made a meta-analysis with a Bayesian fixed effect model in which we related the drug effects to the effects of placebo drugs. We selected 27 clinical trials from the literature according to specific criteria, including results from studies reporting the effects of the newer drugs when tested against diuretics and beta-blockers, and from studies in which diuretics and beta-blockers had been tested against placebo. We calculated the posterior probability distributions of the relative effects of angiotensin-converting enzyme (ACE) inhibitors vs calcium antagonists with three different endpoints: stroke, coronary disease and heart failure with point estimates of effects and with 95% credibility intervals. As an intermediate step in this procedure we obtained similar information about the effects of the three groups of active drugs, ACE inhibitors, calcium antagonists and diuretics or beta-blockers, tested against placebo. For coronary disease we also tested calcium antagonists against diuretics or beta-blockers. RESULTS: ACE inhibitors and calcium antagonists have an almost identical ability to prevent stroke in hypertensive individuals with a risk ratio (RR) of 1.04. On the other hand, calcium antagonists reduce coronary disease by only 8% relative to placebo. When ACE inhibitors and calcium antagonists are compared with the Bayesian method, the outcome is a 14% difference in favor of the ACE inhibitors to prevent coronary disease, with a credibility interval almost reaching identity. Nor do calcium antagonists do as well as diuretics or beta-blockers in this respect, RR = 1.12 with 95% credibility interval 1.01-1.24. All the tested drug groups have a profound preventive effect on the occurrence of heart failure when given to hypertensive patients, showing reductions of 42-54%. When ACE inhibitors are compared with calcium antagonists RR = 0.79, with a credibility interval 0.65 0.95. CONCLUSION: There is statistically an indisputable difference between ACE inhibitors and calcium antagonists in respect of effects on coronary disease and heart failure when treating hypertensive individuals, ACE inhibitors being more efficacious. There are no differences in the effect on stroke. Moreover, beta blockers or diuretics are also superior to calcium antagonists in preventing coronary events. PMID- 12775306 TI - Circumflex artery motion; a new angiographic method for assessment of left ventricular function. AB - OBJECTIVE: To evaluate the usefulness of circumflex artery motion (CAM) for assessment of left ventricular (LV) function. DESIGN: Seventy-three consecutive patients referred for coronary angiography and LV angiography were included. Ejection fraction (EF) was calculated from LV angiography and CAM was measured from coronary angiography. RESULTS: The ratio between CAM and the end-diastolic length of the ventricle, which can be denominated long-axis fractional shortening (FS(L)), was found to be a better index of LV function than CAM per se. There was a significant linear correlation between EF and FS(L) (r = 0.81, SEE = 8.2, p < 0.001). When values of FS(L) > or =10% were selected to define a normal EF (> or =50%) there was a sensitivity of 95% and a specificity of 93%. Visual estimation of EF from CAM was not as good as the use of calculated FS(L) but may me useful as a fast screening method. CONCLUSION: LV systolic function can be assessed by studying CAM recorded by coronary angiography. PMID- 12775307 TI - QT dispersion in relation to left ventricular geometry and hypertension in a population study. AB - OBJECTIVE: To study the relationship between QT interval dispersion, arterial hypertension and different left ventricular geometric patterns in the framework of a population study. DESIGN: A random sample of the population of Tallinn, 717 men and women aged 35-59, underwent standard 12-lead ECG at rest and echocardiography. Corrected QT dispersion was considered as prolonged when the duration was > or =70 ms. RESULTS: In hypertensives with concentric and eccentric hypertrophy, the mean values of corrected QT dispersion were significantly higher than in those with normal geometry. In the normotensive group no significant differences of the mean values of corrected QT dispersion were found in relation to left ventricular geometry. Mean values of corrected QT dispersion were higher in hypertensives than in normotensives in each left ventricular geometric pattern. Corrected QT dispersion > or =70 ms was mainly associated with concentric hypertrophy. CONCLUSION: Prolonged corrected QT dispersion is associated with left ventricular geometric abnormalities and arterial hypertension and is mostly related to concentric hypertrophy. PMID- 12775308 TI - Wall motion and perfusion analysis of transmyocardial laser revascularization. AB - OBJECTIVE: Transmyocardial laser revascularization (TMLR) creates channels in the myocardium. The aim of the treatment is to relieve angina in patients with end stage coronary artery disease. We studied the effect of TMLR on myocardial function and perfusion with the combination of cine magnetic resonance imaging (MRI) and thallium scintigraphy. DESIGN: Eight patients with severe triple-vessel coronary artery disease were studied with MRI and thallium scintigraphy before and 6 months after laser treatment. RESULTS: TMLR did not improve global left ventricular (LV) function or myocardial perfusion. However, systolic wall thickening deprived in segments with fixed perfusion defects in 6 months and laser treatment prevented this deprivation (p = 0.03). In addition single photon emission computed tomography (SPECT) imaging indicated that TMLR prevented conversion of reversible into fixed defects. CONCLUSION: In severe, progressing coronary artery disease TMLR does not improve global LV function or myocardial perfusion, but it preserves systolic wall thickening in fixed defects (scar). It also prevents changes from ischemic myocardial regions to scar. PMID- 12775309 TI - Magnetic resonance imaging of patients with increased blood pressure and altered blood pressure response to exercise after coarctation repair. AB - OBJECTIVE: Patients successfully operated for coarctation of the aorta are frequently subjected to altered blood pressure (BP) at rest and BP response during exercise. The relationship between these variables and blood flow, peak velocity, restenosis and other morphological features of the thoracic aorta as revealed by magnetic resonance imaging (MRI) was evaluated. DESIGN: Fifty-one patients subjected to coarctectomy of the aorta were examined by MRI. In addition, a control group of 23 healthy volunteers was evaluated. Morphology of the aorta was demonstrated with both ECG-triggered SE imaging and gadolinium enhanced MR aortography. Flow-weighted MRI was applied for quantitative flow and velocity measurements. RESULTS: Structural alteration of the aorta was more commonly seen in those patients having increased BP at rest or altered BP response during exercise than those with a normal BP profile. The luminal diameter of the narrowest site of the aorta was decreased in all patient groups. Accordingly, the peak velocity at the corresponding site was significantly (p < 0.01) increased. However, blood flow was significantly (p < 0.01) decreased among those patients with normal BP profile compared with the other patient groups as well as the controls. CONCLUSION: Other structural changes than restenosis may contribute as well to the altered BP profile of patients subjected to coarctectomy. Reduced blood flow appears to correlate with normal BP profile, whereas the peak velocity measurements that are obtained by MRI are not able to differentiate between the patient groups. The comprehensive and reliable data obtained by non-invasive techniques, i.e. MRI and Doppler, may replace catheterization when deciding the need for intervention. PMID- 12775310 TI - Excellent survival and low complication rate in medium-term follow-up after arterial switch operation for complete transposition. AB - OBJECTIVE: To determine medium-term outcome after arterial switch operation for complete transposition in a population-based cohort. DESIGN: Retrospective study of all 86 patients operated on at our institution from 1982 to 2000. Age at operation was 0-256 days (median 6 days); 28 (32%) had closure of a ventricular septal defect, 2 of whom had double outlet right ventricle. Additional surgery included coarctectomy in three, pulmonary artery banding in two and atrial septectomy in one patient. RESULTS: Thirty-day operative mortality was 7/86 (8.1%), after 1988 mortality was only 3.8% and confined to patients with unusual coronary artery patterns. There was no late mortality; Kaplan-Meier survival was 91%. Five patients (6% of survivors) were reoperated for right ventricular outflow tract obstruction. At last follow-up, 86% were free from complications; three had neurological complications, two were on treatment for ventricular dysfunction and one had a pacemaker for postoperative atrioventricular block. CONCLUSION: In this cohort of first-generation patients treated with arterial switch operation, medium-term survival is excellent and the rate of reoperation and sequelae is low. PMID- 12775311 TI - Haemodynamic adaptation during exercise in fontan patients at a long-term follow up. AB - OBJECTIVE: To assess the haemodynamic regulation at rest and during exercise in Fontan patients at a long-term follow-up. DESIGN: Cardiac output was measured with the dye-dilution technique. We examined 15 out of the surviving 20 patients operated upon in Goteborg between 1980 and 1991. Their mean age was 26.4 years. Four patients had to be excluded due to technical reasons. RESULTS: Median maximal oxygen uptake was 1.47 l/min, corresponding to 21.9 ml/kg/min. Cardiac output was lower than expected at all exercise levels, presumably due to a reduced pulmonary blood flow. The median maximal cardiac output value was 8.0 l/min. Stroke volume index was 33 ml/m (2). The subjects compensated for the reduced cardiac output with an increased arteriovenous oxygen difference. They had a normal increase in arterial blood pressure. This was achieved by an increase in total peripheral resistance. CONCLUSION: The low maximal exercise capacity was due to a reduced cardiac output and a reduced pulmonary blood flow. This was compensated for by an increased arteriovenous oxygen difference PMID- 12775312 TI - Myocardial uptake and release of substrates in patients operated for unstable angina: impact of glutamate infusion. AB - OBJECTIVE: To study cardiac uptake and release of substrates and the influence of intravenous glutamate in patients operated for unstable angina requiring intravenous nitrates. DESIGN: Nineteen patients were randomized to blinded infusion of glutamate or saline. Arterial-coronary sinus differences of substrates were measured before cardiopulmonary bypass (CPB) and during early reperfusion. RESULTS: Before CPB the only major substrates that were extracted by the heart in the saline group were free fatty acids (FFAs). During reperfusion uptake of glucose and glutamate was found but FFAs remained the major substrate extracted by the heart. Initially transient low oxygen extraction and lactate release were found. Conversion to lactate uptake was not observed. Glutamate infusion was associated with an uptake of glutamate and in contrast to the control group there was also uptake of lactate before CPB and at the end of the study period. CONCLUSION: The metabolic situation before CPB with a reliance on myocardial FFA uptake is less than ideal with regard to ischemia. Early reperfusion was characterized by dynamic changes and a shift towards myocardial glucose uptake but FFAs remained the major substrate extracted. The qualitative findings associated with glutamate infusion agree with previous animal and human studies but have to be interpreted cautiously due to lack of flow measurements PMID- 12775313 TI - Placebo effects: artifact of modern clinical trial conduct. PMID- 12775314 TI - Eumovate (clobetasone butyrate 0.05%) cream: a review of clinical efficacy and safety. AB - Topical steroid creams and ointments have been available as over-the-counter (OTC) medications for the self treatment of acute dermatitis and other steroid responsive skin disorders for more than ten years. Despite earlier fears, widespread availability and use of these creams is not associated with clinically significant adverse effects. In dermatological practice, hydrocortisone 1% remains the mainstay of treatment for facial eczema, but it is often not effective in eczema affecting other body areas. Eumovate(TM) (clobetasone butyrate 0.05%) cream has recently been made available as a pharmacy medication for the short-term management of acute eczema and allergic dermatitis by adults and children aged 10 or older, based on evidence derived from clinical trials involving over 3500 patients. This review summarises the key efficacy and safety data derived from 29 clinical trials and the post-licensing pharmacovigilance safety information, which supported the reclassification of this product for OTC use. These data show clobetasone butyrate 0.05% is more effective than 1.0% hydrocortisone in the treatment of eczema and more effective than flurandrenolone 0.0125% (p=0.01%) and a potent topical steroid hydrocortisone butyrate (p<0.05), in the treatment of psoriasis. A review of the effect of topical steroids on skin thickness concluded that, following short term application, there was no clinically significant difference between hydrocortisone 1.0% and clobetasone butyrate 0.05% in terms of potential for skin thinning. Similarly, even under extreme conditions, clobetasone butyrate 0.05% has negligible systemic absorption and has almost no effect on HPA axis function. PMID- 12775315 TI - Safe treatment of head/neck AD with tacrolimus ointment. AB - BACKGROUND: Atopic dermatitis(AD) with head and neck involvement is common and therapeutically challenging. METHODS: Efficacy and safety data specific to treatment of head/neck regions with tacrolimus ointment (Protopic) from three double-blind, randomized, vehicle-controlled studies are reported. A total of 631 adult and 352 pediatric patients with moderate to severe atopic dermatitis applied the vehicle, 0.03% or 0.1% tacrolimus ointment twice daily to affected areas for up to 12 weeks. RESULTS: Significant improvements from baseline to end of treatment for signs of atopic dermatitis (erythema, edema, excoriation, oozing, scaling, and lichenification) were noted for head/neck and non-head/neck areas treated with either 0.03% or 0.1% tacrolimus ointment (p<0.001). Within each treatment group, the overall 12-week adjusted incidence rate of application site adverse events was similar for both head/neck and non-head/neck areas. The incidence of common adverse events such as pruritus, "skin burning", erythema, infection, and skin tingling in head/neck areas was comparable to that observed in non-head/neck areas within each treatment group. The overall prevalence of application site adverse events decreased rapidly during the first few days of treatment. CONCLUSION: Tacrolimus ointment is a safe and effective treatment for atopic dermatitis on the head and neck. PMID- 12775316 TI - Itraconazole pulse therapy improves the quality of life of patients with toenail onychomycosis. AB - BACKGROUND: Toenail onychomycosis is a common disease that can have serious adverse effects on the quality of life (QOL) of patients. AIM: To evaluate the impact of itraconazole pulse therapy on the QOL of patients with toenail onychomycosis. METHODS: A total of 20 patients with disto-lateral subungual toenail onychomycosis were treated with itraconazole 200 mg twice daily for 1 week every 4 weeks for 12 weeks. The patients were asked to complete a QOL questionnaire before treatment and on the last follow-up visit (week 48). A score of 0-4 was given according to the five possible responses to each question and these were summed to give the final score of the patient. The mean of the final scores of the patients before and after treatment were compared using the Wilcoxon matched-pairs test. RESULTS: At 48 weeks after commencing treatment, 14 patients (70%) responded to treatment (nine patients were cured with almost totally clear toenails and five patients improved), and 16 patients (80%) were mycologically cured (negative KOH smear and culture). The mean of the QOL scores of the patients before treatment was 18.0+/-7.8, which reduced to 13.1+/-11.3 after treatment (two-tailed, p=0.009). CONCLUSION: Itraconazole pulse therapy is an effective treatment and can improve the QOL of patients with toenail onychomycosis. PMID- 12775317 TI - A comparison of photodynamic therapy using topical methyl aminolevulinate (Metvix) with single cycle cryotherapy in patients with actinic keratosis: a prospective, randomized study. AB - BACKGROUND: Actinic keratosis (AK) is a very common condition, which has the potential of progressing to squamous cell carcinoma. The present study is a prospective, randomized study comparing the lesion response, cosmetic outcome, patient satisfaction and tolerability of a new treatment modality, photodynamic therapy (PDT), using topical methyl aminolevulinate (Metvix), with the most commonly used standard therapy for AK, cryotherapy. METHODS: A total of 204 patients with clinically diagnosed AK were randomized to either cryotherapy or PDT. The PDT patients were further assigned to an active or placebo group in a random, double-blind manner. Cryotherapy was performed using liquid nitrogen spray in a single freeze-thaw cycle. PDT was performed using 160 mg/g methyl aminolevulinate cream or placebo, a 3-hour application time, red light (570-670 nm) and a total light dose of 75 J/cm(2). PDT was repeated after 7 days. Two sessions of PDT were undertaken, as a previous study had shown a single session had similar efficacy to cryotherapy. Lesion response was assessed clinically after 3 months (complete response or non-complete response). RESULTS: The lesion response rate was 91% in the methyl aminolevulinate PDT group, 68% in the cryotherapy group and 30% in the placebo PDT group. Methyl aminolevulinate PDT was statistically significantly better than both cryotherapy and placebo PDT in terms of response rates and cosmetic outcome. Most patients preferred PDT to other treatments. CONCLUSIONS: PDT with methyl aminolevulinate is an excellent treatment option, particularly for patients with widespread damage or AK lesions in cosmetically sensitive areas. PMID- 12775318 TI - Photosensitizers for photodynamic therapy of cutaneous disease. AB - Photodynamic therapy is a rapidly developing treatment modality in dermatology. A sensitizer drug is activated by light in the presence of oxygen. This results in the release of reactive oxygen species that damage the target tissue. The ideal features of a photosensitizer are that it should be highly selective for lesional tissue, activated by light of a sufficiently long wavelength for tissue penetration, and have a high photodynamic yield (i.e. production of singlet oxygen). A short time interval between administration and its maximal accumulation in tumour tissue, followed by rapid tissue clearance, are also desirable. First-generation sensitizers were complex chemical mixtures, needing parenteral administration and causing troublesome and prolonged photosensitivity. A range of second-generation sensitizers of different chemical families show several advantages including purity, longer activation wavelength and less prolonged photosensitivity, but effective topical formulations have not been developed. Currently, the most convenient and widely practised form of PDT for cutaneous disorders is the topical application of the pro-drug delta aminolevulinic acid or its methylated ester, which are activated by light following metabolism to the endogenous sensitizer protoporphyrin IX. PMID- 12775319 TI - Cyclosporin in childhood psoriasis. AB - BACKGROUND: Cyclosporin is known to be highly effective in the treatment of psoriasis in adults. It has also proved effective and well tolerated in the treatment of severe childhood atopic dermatitis. Psoriasis in childhood is relatively unusual but by no means rare and on occasions the disease can be very difficult to control in this age group. The use of cyclosporin for psoriasis in childhood has received scarcely any attention and in the few cases that have been reported the results have been inconsistent. Three children aged from 7 to 11 years with severe psoriasis resistant to topical agents were treated with cyclosporin. The highest dose required was 3.5 mg/kg per day. The duration of treatment ranged from 6 weeks to 4 months. Cyclosporin was effective and generally well tolerated. Treatment was interrupted in one case due to nausea and diarrhoea. None of the patients developed hypertension or renal impairment. The potential role of cyclosporin in severe childhood psoriasis is discussed. PMID- 12775320 TI - Radiotherapy for in situ extramammary Paget disease of the vulva. AB - BACKGROUND: Radiotherapy as a first-choice treatment for in situ extramammary Paget disease has been successfully used. OBJECTIVES: To review the most relevant aspects of radiotherapy as first-choice treatment in selected cases of in situ extramammary Paget disease of the vulva. PATIENTS AND METHODS: Two Caucasian females aged 76 and 92 years with in situ extramammary Paget disease localized in the genital region were treated by means of ortovoltage X-rays: 100 kV, 8 mA, 1.7 mm Al filter, field size of 12-cm cone, and source skin distance of 30 cm. Both patients received 40 Gy, 200 cGy per fraction, five fractions per week. RESULTS: Complete regression of in situ extramammary Paget disease was observed in both patients after radical radiation therapy and neither local recurrences nor internal malignancies were detected. CONCLUSIONS: Radiotherapy is a curative treatment in selected cases of in situ extramammary Paget disease affecting the vulva. PMID- 12775321 TI - Severe erythema nodosum due to Behcet's disease responsive to erythromycin. AB - A patient with severe erythema nodosum due to Behcet's disease is reported on here. Erythema nodosum lesions did not respond to classical treatments; however, they cleared after erythromycin treatment, which was prescribed for the treatment of coincidental erythrasma. Erythromycin treatment appears to be an effective treatment option in erythema nodosum. The hypothetical anti-inflammatory effects of erythromycin, besides its antibiotic properties, are reviewed and discussed to explain such a clinical improvement. PMID- 12775326 TI - Animal models of anxiety. PMID- 12775327 TI - The Syracuse strains, selectively bred for differences in active avoidance learning, may be models of genetic differences in trait and state anxiety. AB - The derivation of the Syracuse high- and low-avoidance strains is described. The behavioral characterization of the high- and low-avoidance phenotypes is summarized and it is concluded that the SLA/Bru strain is best described as having higher state and trait anxiety than their SHA/Bru counterparts. Although the behavioral covariates of the high- and low avoidance phenotypes are consistent, the covariation of the endocrine system normally thought to be involved in stress, is anomalous. The SLA/Bru rats, which are behaviorally more anxious than the SHA/Bru animals, show hypertrophy of the adrenal glands but reduced synthesis and release of the stress-related corticosterone than the SHA/Bru animals. This dissociation of the behavioral and endocrine measures of anxiety appears to be genetic, since a selective genetic analysis, involving F2 and high and low backcross segregating generations, indicates that both the behavioral and endocrine covariates cosegregate with the avoidance phenotypes. These data suggest that the expected association of behavioral and endocrine measures of anxiety is correlational, not causal. PMID- 12775328 TI - Divergent stress responses and coping styles in psychogenetically selected Roman high-(RHA) and low-(RLA) avoidance rats: behavioural, neuroendocrine and developmental aspects. AB - The Swiss sublines of Roman high-(RHA/Verh) and low-(RLA/Verh) avoidance rats have been genetically selected for good vs. poor performance in two-way active avoidance since 1972. RLA/Verh rats show increased stress responses (e.g. freezing behaviour, ACTH, corticosterone and prolactin secretion) and adopt a more passive (or reactive) coping style when confronted with a novel environment. In the open field, elevated plus-maze, black/white box test, and in a new light/dark open field test, RLA/Verh rats appear to be more anxious than their RHA/Verh counterparts. Anxiety may result from their particular psychophysiological profile, i.e. increased emotionality combined with a passive coping style. In contrast, RHA/Verh rats are less responsive to stress, they show little anxiety in novel situations and tend to be impulsive and novelty (sensation) seekers. Some behavioural differences are already noticeable shortly after birth, but the full pattern appears to stabilize only after puberty. Gene environment interactions are critical in establishing this pattern. The data reviewed indicate that the differences between RHA/Verh and RLA/Verh rats probably result from a complex interaction among divergent anxiety/emotionality characteristics, differences in locomotor activity and novelty/reward seeking, as well as active vs. passive coping styles. It is proposed further that these divergent personality types are to be found not only in other selective breeding programs but in the form of individual differences in most populations of rats used for this type of research. PMID- 12775329 TI - Involvement of 5-HT1A receptors in animal tests of anxiety and depression: evidence from genetic models. AB - Clinical studies have suggested the involvement of 5-HT1A receptors in anxiety and depressive disorders because partial 5-HT1A receptor agonists such as buspirone are therapeutic. The present review considers evidence from genetic animal models that support a role for 5-HT1A receptors in anxiety-like and depressed-like behavior in animals. Selective breeding for differential hypothermic responses to a selective 5-HT1A receptor agonist led to the development of the high DPAT sensitive (HDS) and low DPAT sensitive (LDS) lines of rats. The HDS rats differ from the LDS rats on several behavioral measures reflective of anxiety or depression, including reduced social interaction, reduced responding in a conflict task and exaggerated immobility in the forced swim test. However, they do not differ from the LDS rats in the elevated plus maze task, which is a commonly used test of anxiety. Nor do the HDS rats exhibit a typical anxiogenic response to the hippocampal administration of the 5-HT1A agonist. Although the HDS rats do exhibit elevations in 5-HT1A receptors in regions of the limbic cortex, it is not clear whether these increases account for the behavioral differences. Paradoxically, 5-HT1A receptor knockout mice also exhibit anxiety-like behavior in the plus maze, open field and conflict tests compared to wild type mice. However, the knockouts exhibited less immobility in the forced swim test than wild type control mice. Recent studies using selective regional reinstatement of the receptor have implicated the postsynaptic 5-HT1A receptors in these changes in anxiety-like behavior. Thus, preliminary evidence from two different types of genetic animal models suggests that anxiety-like behavior can arise if the 5-HT1A receptor function is eliminated or overexpressed. Further study with additional tests of anxiety are needed to confirm this intriguing relationship. PMID- 12775330 TI - Born to be anxious: neuroendocrine and genetic correlates of trait anxiety in HAB rats. AB - This review summarises behavioural, neuroendocrine, and genetic characteristics of Wistar rats bred for either high (HAB) or low (LAB) anxiety-related behaviour. Compared to LABs, HAB animals show signs of extreme trait anxiety in a variety of behavioural tests; they further prefer passive coping strategies, indicative of a genetically linked depression-like behaviour, and show signs of increased stress vulnerability. All behavioural parameters associated with trait anxiety are robust and consistent. Resembling psychiatric patients, HAB rats respond to exposure to ethologically relevant stressors with a hyper-reactivity of the hypothalamic-pituitary-adrenal axis and show a pathological outcome of the combined dexamethasone/corticotropin-releasing hormone (Dex/CRH) challenge test. Experimental evidence indicates that over-expression and -release of vasopressin in the hypothalamic paraventricular nucleus is responsible for these behavioural and neuroendocrine phenomena, making the neuropeptide gene a candidate gene of trait anxiety/depression. Indeed, preliminary molecular genetic approaches succeeded in identifying polymorphisms in the promoter structure of the vasopressin gene. This may have implications for understanding the molecular basis for individual variations in trait anxiety and for psychopathology. PMID- 12775331 TI - Dissecting adrenal and behavioral responses to stress by targeted gene inactivation in mice. AB - To define the molecular pathways modulating adrenal and behavioral responses to stress, we have generated mice with inactivation of hypothalamic neuropeptides and signaling pathways. Studies in mice deficient in corticotropin-releasing hormone (CRH) have revealed the essential role for CRH in adrenal glucocorticoid production in response to many physiological and psychological stressors. Immune system activation in CRH-deficient mice provides a unique exception to the necessity for CRH in stimulating adrenal glucocorticoid production. By analyzing mice deficient in interleukin-6 (IL-6) and CRH, we find that restoration of glucocorticoid output with inflammation is largely mediated by dysregulated IL-6 production. Current studies focus on identifying cellular and gene targets by which glucocorticoids regulate immune system function. In contrast to impaired adrenocortical responses to stress, CRH-deficient mice exhibit normal behavioral responses to stress. To determine signaling pathways that may contribute to the behavioral responses to stress, we have generated and analyzed mice deficient in adenylyl cyclase type 8 (AC8). AC8 deficient mice have intact adrenocortical responses to stress, but an inability to undergo stress-induced alterations in behavior. PMID- 12775332 TI - Effect of swimming session duration and repetition on metabolic markers in rats. AB - The aim of this study was to investigate the profile of metabolites in male rats subjected to 50-60 min of swimming on three protocols: group A, a single 50 min swimming session; group B, one session a day for three days (5 min on day 1, 15 min on day 2 and 30 min on day 3); and group C, one session a day for 5 days, with increasing duration from 5 min on day 1, 15, 30, 45 and 60 min on consecutive days. The interval between sessions was 24 h. Measurements were made after the last swimming session. Controls did not swim. The glycogen content of liver and gastrocnemius and soleus muscle was depleted in the three groups that swam, but blood glucose concentration was significantly increased only in group B. Serum lactate concentrations were greater than the controls in groups A and B. There were significant increases in serum free fatty acid concentrations in all groups that swam. The increases in plasma free fatty acids may have resulted from lipolysis stimulated by endogenous catecholamines in groups A and C, since basal lipolysis measured in vitro was unchanged by swimming. The large increase in basal lipolysis in group B may have contributed to the rise in plasma free fatty acids. Adipocytes from rats in groups A and B were supersensitive to epinephrine, whereas those from group C were not. We conclude that the metabolic alterations were less pronounced after the last of five swimming sessions over 5 days than after a single session, even though session duration and the contribution of the physical component were similar. Glucose mobilization, but probably not utilization, was similar in the three groups that swam. The mechanisms of lipid mobilization from adipose tissue differed, depending on the stress paradigm. The metabolic changes in groups A and B indicated that three daily swimming sessions were insufficient to cause adaptation. The results contrast with previous findings for foot-shock stress, which leads to sensitization rather than adaptation in response to repeated stimuli. PMID- 12775333 TI - Gender differences in cardiovascular and hypothalamic-pituitary-adrenal axis responses to psychological stress in healthy older adult men and women. AB - Gender differences in the neuroendocrine and cardiovascular response to psychological stress may contribute to the gender differences in the prevalence of diseases associated with hypothalamic-pituitary-adrenal (HPA) axis reactivity such as cardiovascular disease (CVD), diabetes and hypertension. We measured plasma ACTH, cortisol, heart rate (HR), and blood pressure (BP) responses in 8 men and 8 women (55-75 years) exposed to the Matt Stress Reactivity Protocol (MSRP), a psychological challenge. The MSRP elicited significant increases in HR, systolic-, and diastolic BP, ACTH and cortisol (all p<0.01). Men had significantly greater cortisol and diastolic BP responses compared to women (p<0.05). Additionally, a positive correlation between the ACTH and cortisol responses was only found in the males (r=0.71, p<0.05). There were no group differences in HR, systolic BP, or ACTH responses. We conclude, that among older adults, men respond to psychological stress with greater increases in cortisol, compared to women. This greater activation of the HPA axis could translate into an elevated risk for CVD, diabetes and hypertension and may be related to the higher prevalence of these diseases in males. Gender differences in brain structures and/or cognitive processes may be responsible for these sexually dimorphic stress responses. PMID- 12775334 TI - Enhanced recovery of NK cell activity in mice under restraint stress by the administration of a biological response modifier derived from the mycelia of the basidiomycete Tricholoma matsutake. AB - Some types of stressor act on the immune system via the network comprising the endocrine-immune-nervous systems, and are reportedly responsible for the onset of diseases as well as giving impetus to their advance. It is important for the maintenance and promotion of health to cope with stress-induced changes in immunocompetence. Therefore, we studied the effects of administration of a novel biological response modifier (CM6271) derived from the mycelia of the basidiomycete Tricholoma matsutake on the NK cell activity in mice under restraint stress, in order to evaluate its potential to modulate immune responsiveness in stress-loaded individuals. (1) When C57BL/6 mice were restrained in 50-ml tubes for more than 6 h, splenic NK cell activity decreased significantly, but recovered gradually after the mice were released. The extent of the reduction of activity and the degree of recovery depended on the duration of the restraint. (2) The oral administration of CM6271 caused a significant acceleration of the recovery of the activity. This effect was dependent on the timing of administration and the dose given. (3) The administration of CM6271 had no clear effect on the blood levels of ACTH, corticosterone or lipid peroxide levels in the liver. These findings suggest that CM6271 promotes recovery from the decrease in NK cell activity induced by restraint stress. PMID- 12775336 TI - Activity, regulation, and intracellular localization of RGS proteins. AB - RGS proteins attenuate the activities of heterotrimeric G proteins largely by promoting the hydrolysis of the activating nucleotide GTP. This review discusses the interactions of RGS proteins and G proteins and how those interactions are regulated by a variety of factors including auxiliary proteins and other cellular constituents, posttranslational modifications, and intracellular localization patterns. In addition, we discuss progress that has been made toward understanding the roles that RGS proteins play in vivo, and how they may serve to govern responses to G protein-coupled receptors upon acute and prolonged activation by agonists. PMID- 12775337 TI - Genetic approaches to visual transduction in Drosophila melanogaster. AB - Because almost everything we know about Drosophila phototransduction has come from studies based on genetic approaches, this review begins with a discussion of genetic approaches. We then present a brief overview of Drosophila phototransduction (section on Drosophila phototransduction: an overview) followed by a more detailed treatment of individual components of the transduction machinery (section on Components of the phototransduction machinery). Discussion of transduction mechanisms is presented under three headings: Mechanism(s) of channel excitation, Organization of the transduction proteins, and Regulatory mechanisms in phototransduction. Perhaps the most important unanswered question in this field is the mechanism(s) of activation and regulation of transduction channels. This question is explored in the section entitled Mechanism(s) of channel excitation. Identification of at least two of the proteins discussed was totally unexpected: the rhodopsin chaperone protein, ninaA, and the signal complex scaffold protein, INAD. They are discussed in the sections titled Requirement for a chaperone protein for Rh1 opsin, and: Formation of signaling complexes, respectively. One of the important developments in this field has been the discovery of mammalian homologs of many of the proteins identified in Drosophila. A brief discussion of the most extensively studied of these, the mammalian homologs of light-activated channel protein, trp, is presented in the section on Mammalian Homologs of trp. We conclude the review with Perspective, a brief look at the current status and the future outlook of the field. PMID- 12775338 TI - Protein complexes involved in heptahelical receptor-mediated signal transduction. AB - Signal transduction mediated by heterotrimeric G proteins that couple to heptahelical receptors requires the involvement of many different proteins. Although some of the early evidence suggested that signal transduction components were assembled into complexes, much of the data supported an alternative hypothesis positing that the process involved transient interactions driven by random collision events. However, recent data indicate that many of the components involved in signal transduction do indeed form complexes. Here we review the evidence for these complexes and how they contribute to the specificity and efficiency of signaling in cells that must manage numerous signal transduction pathways. PMID- 12775339 TI - Accessory proteins for G protein-signaling systems: activators of G protein signaling and other nonreceptor proteins influencing the activation state of G proteins. AB - Heterotrimeric G proteins are key transducers for signal transfer from outside of the cell. In addition to their regulation by the superfamily of G protein-coupled receptors, many if not all of the subtypes of heterotrimeric G proteins are also regulated by additional accessory proteins that influence guanine nucleotide binding and/or hydrolysis or subunit interactions. Activators of G protein signaling (AGS1-3) refer to a functionally defined group of proteins that activate G protein-signaling systems in the absence of a classical G protein coupled receptor. AGS and related proteins provide unexpected insights into the regulation of the G protein activation/deactivation cycle and the functional roles of G proteins. These proteins likely play important roles in the generation of signaling complexes, the positioning of signaling proteins within the cell, and in biological roles of G proteins unrelated to a cell surface receptor. As such, these proteins and the concepts advanced with their discovery provide unexpected avenues for therapeutics and understanding disease mechanisms. PMID- 12775340 TI - A novel kind of G protein heterodimer: the G beta5-RGS complex. AB - The fifth member of the G protein beta the subunit family, G beta5, has been shown to bind exclusively to a subfamily of regulators of G protein signaling (RGS) including RGS6, RGS7, RGS9, and RGS11. This interaction occurs through a G protein gamma-like (GGL) domain present in members of this RGS subfamily and is the only reported instance in which a G beta subunit is not bound to a G gamma subunit. The G beta5-RGS interaction has been demonstrated both in vitro and in vivo and has been shown to stabilize the dimer against proteolytic degradation. GTPase activating protein (GAP) assays suggest that G beta5-RGS7 acts specifically on G alphao, however in cell-based assays it also inhibited G alphai and G alphaq-mediated signaling. The role of the dimer in signaling and the function of G beta5 moiety within the complex are poorly understood. This review summarizes the information about the assembly and function of G beta5-RGS dimers, as well as their posttranslational modifications and localization. PMID- 12775341 TI - Spinal cord injury: inductive lability can enhance and hasten recovery. AB - In spinal cord injury, recovery of function, if any, confirms the presence of survived neural tissue at the injury site. However, recovery several years after the injury remains unexplained. Body weight bearing locomotor exercises seem to bring these new outcomes. Developing locomotor system and computer simulation studies show that motor learning requires the presence of redundant sets of competing synapses within the spinal cord interneurons. The new exercise regimens have not addressed this essential prerequisite; this could perhaps explain the long delays in recovery. We recommend that inclusion of inductive lability procedure (Krishnan, 1983, 1991, 2003) will help hasten and enhance the recovery. PMID- 12775342 TI - Connexin 32 mutation in a Turkish family with X-linked Charcot-Marie-Tooth disease. AB - In the present work, we describe a large Turkish family (N=39) with Charcot-Marie Tooth disease, which is the most commonly inherited peripheral neuropathy. The subjects were from four generations, including six hemizygote patients and nine heterozygote carrier females. Symptoms appeared in late childhood in males (mean age=13.5) but later in carrier females (mean age=33.5). The peripheral nerve conduction was more severely affected in males than females. Genomic DNA was isolated from peripheral white blood cells. Using SSCP technique (single strand conformation polymorphism analysis), abnormal patterns of migration were observed in 15 subjects: 6 of them were hemizygote males and 9 were heterozygote carrier females. We identified a mutation of the Cx32 gene, consisting of a guanine to adenine transition at position 271 (271G-A). The results suggested relations between degenerative processes and position of Cx32 mutations. PMID- 12775343 TI - Tactile stimulus and neurohormonal response: a pilot study. AB - The effects of tactile stimuli on plasma oxytocin and neuropeptide Y (NPY) were investigated in 21 volunteers exposed to massage. Blood samples for basal values were drawn immediately before and immediately after finishing the massage. A third sample was drawn after 60 min of restricted rest. On focusing on the difference between oxytocin concentrations before and immediately after massage, we found a sex difference. An opposite sex difference was found for NPY. The results imply that there might be sex-related difference in neurohormonal response to tactile stimuli such as in massage, and the results contradict those of previously reported animal experiments. PMID- 12775344 TI - Hypothalamic digoxin and hemispheric chemical dominance: relation to speech and language dysfunction. AB - The isoprenoid pathway produces three key metabolites--endogenous digoxin, dolichol, and ubiquinone. Since endogenous digoxin can regulate neurotransmitter transport and dolichols can modulate glycoconjugate synthesis important in synaptic connectivity, the pathway was assessed in patients with dyslexia, delayed recovery from global aphasia consequent to a dominant hemispheric thrombotic infarct, and developmental delay of speech milestone. The pathway was also studied in right hemispheric, left hemispheric, and bihemispheric dominance to find out the role of hemispheric dominance in the pathogenesis of speech disorders. The plasma/serum--activity of HMG CoA reductase, magnesium, digoxin, dolichol, ubiquinone--and tryptophan/tyrosine catabolic patterns, as well as RBC (Na+)-K+ ATPase activity, were measured in the above mentioned groups. The glycoconjugate metabolism and membrane composition was also studied. The study showed that in dyslexia, developmental delay of speech milestone, and delayed recovery from global aphasia there was an upregulated isoprenoidal pathway with increased digoxin and dolichol levels. The membrane (Na+)-K+ ATPase activity, serum magnesium and ubiquinone levels were low. The tryptophan catabolites were increased and the tyrosine catabolites including dopamine decreased in the serum contributing to a speech dysfunction. There was an increase in carbohydrate residues of glycoproteins, glycosaminoglycans, and glycolipids levels as well as an increased activity of GAG degrading enzymes and glyco hydrolases in the serum. The cholesterol:phospholipid ratio of RBC membrane increased and membrane glycoconjugates showed a decrease. All of these could contribute to altered synaptic inactivity in these disorders. The patterns correlated with those obtained in right hemispheric chemical dominance. Right hemispheric chemical dominance may play a role in the genesis of these disorders. Hemispheric chemical dominance has no correlation with handedness or the dichotic listening test. PMID- 12775345 TI - A hypothalamic digoxin-mediated model for conscious and subliminal perception. AB - The human hypothalamus produces an endogenous membrane (Na+)-K+ ATPase inhibitor digoxin. Digoxin can modulate multiple neurotransmitter systems. It can play a role in perceptual binding, focused attention and short term memory important in conscious perception. It can also mediate subliminal or quantal perception. A hypothalamic digoxin-mediated model for conscious and quantal perception is postulated. PMID- 12775346 TI - Hypothalamic digoxin, regulation of neuronal transmission, and cerebral dominance. AB - The present study assessed the neurochemical differences between right hemispheric dominant and left hemispheric dominant individuals. The HMG CoA reductase activity, serum digoxin, magnesium, tryptophan catabolites, tyrosine catabolites, and RBC membrane (Na+)-K+ ATPase activity were measured in individuals of differing hemispheric dominance. The results showed that right hemispheric dominant individuals had elevated digoxin synthesis, increased tryptophan catabolites, and reduced tyrosine catabolites and membrane (Na+)-K+ ATPase with hypomagnesemia. Left hemispheric dominant individuals had the opposite patterns. Right hemispheric dominance represents a hyperdigoxinemic state with membrane sodium-potassium ATPase inhibition. Left hemispheric dominance represents the reverse pattern with hypodigoxinemia and membrane sodium potassium ATPase stimulation. PMID- 12775347 TI - Quantitative EEG analysis in obsessive compulsive disorder. AB - Quantitative analysis of the EEG (q-EEG) in patients with obsessive compulsive disorder (OCD) showed a decreased beta and an increased theta power at frontotemporal regions. The patients who had higher scores in doubting test (Maudsley Obsessive Compulsive Questionnaire) and more severely ill patients shared similar q-EEG features. The relative theta powers were significantly increased and alpha powers were significantly decreased in these patients, particularly in the frontotemporal region. It was suggested that the q-EEG may be useful in investigating the OCD patients with heterogeneous characteristics. PMID- 12775348 TI - Line bisection task performance and resting EEG alpha power. AB - Neurologically normal subjects generally err to the left of veridical center when performing a line bisection task, a phenomenon termed "pseudoneglect." We hypothesized that resting electroencephalogram (EEG) alpha oscillations may show relationships with attentional mechanisms and give some clues about the underlying mechanisms of pseudoneglect. We recorded resting EEGs of 41 subjects and tested them with a paper-pencil line bisection task. Our results showed that line bisection scores of men (n=18) were less biased and their performance was higher compared to those of women (n=20), but these differences only approached significance. The eyes open resting EEG alpha power of women was significantly and positively correlated with their line bisection performance. In general, significant relationships were related to the left hand performance when the lines were presented in the left hemispace. Greater resting alpha power was correlated with lower absolute bisection score or, in other words, higher bisection performance. Greater alpha power also correlated with diminished leftward bisection bias (or reduced pseudo-neglect). The resting EEG alpha of men was weakly associated with bisection performance. Results discussed in terms of Kinsbourne's activation-orientation theory and Basar's view on brain oscillations. PMID- 12775349 TI - The differential impacts of Alzheimer's dementia, head injury, and stroke on personality dysfunction. AB - The current study was an attempt to empirically measure and compare the personality effects of Alzheimer's dementia, stroke, and head injury. The study hypothesized that there are differences in the Minnesota Multiphasic Personality Inventory (MMPI-2) patterns generated by the three groups. It was hypothesized that head injury shows the most personality dysfunction. The subjects included 124 stroke clients, 290 head injury clients, and 166 dementia clients. These individuals averaged 58.04 years old, while the mean education was 12.67 years with a minimum of 7 years. The individuals were mostly Caucasian, but included 80 African-Americans, Hispanics, or others. The average chronicity was 77.55 months. The dependent variables were the patient's personality characteristics as measured by 15 MMPI-2 scales. A MANCOVA indicated that there was a significant difference among the three groups after covarying for age, education, and sex (F(30, 1116)=11.03, p<.001). There were overall differences among 10 of the 15 MMPI scales according to univariate follow-up tests. The major pattern was increased pathology for the Head Injury Group. The Stroke and Dementia Groups differed from each other on 6 of the 15 scales but the overall level of severity was similar. High scores on depression and schizophrenia were seen in all three groups. The data appeared to suggest that such factors as the speed of onset of the injury as well as organic factors both played a role in determining personality dysfunction. The elevation on schizophrenia reflected both personality dysfunction as well as a general elevation related to brain injury. PMID- 12775350 TI - Repetitive behaviors in chronic schizophrenia: using the Japanese version of the Elgin Behavior Rating Scale (JEBRS). AB - The main aim of this study was to test the reliability and value of the Japanese Elgin Behavior Rating Scale (JEBRS) with 92 schizophrenia patients. Cronbach's alpha for the JEBRS was 0.61. All inter-rater reliability coefficients were in the satisfactory range. The JEBRS had a high stability over time. Almost all of the correlations among the nine item scores and total score in the JEBRS were significant. The total score for nine repetitive behaviors in the JEBRS was positively correlated with positive and negative symptoms. This study showed that the JEBRS is a useful and reliable scale. PMID- 12775356 TI - Dexamethasone prevents granulocyte-macrophage colony-stimulating factor-induced nuclear factor-kappaB activation, inducible nitric oxide synthase expression and nitric oxide production in a skin dendritic cell line. AB - AIMS: Nitric oxide (NO) has been increasingly implicated in inflammatory skin diseases, namely in allergic contact dermatitis. In this work, we investigated the effect of dexamethasone on NO production induced by the epidermal cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF) in a mouse fetal skin dendritic cell line. METHODS: NO production was assessed by the method of Griess. Expression of the inducible isoform of nitric oxide synthase (iNOS) protein was evaluated by western blot analysis and immunofluorescence microscopy. Western blot analysis was also performed to evaluate cytosolic IkappaB-alpha (IkappaB alpha) protein levels. The electrophoretic mobility shift assay was used to evaluate the activation or inhibition of nuclear factor kappa B (NF-kappaB). RESULTS: GM-CSF induced iNOS expression and NO production, and activated the transcription factor NF-kappaB. Dexamethasone inhibited, in a dose-dependent manner, NO production induced by GM-CSF. Addition of dexamethasone to the culture, 30 min before GM-CSF stimulation, significantly inhibited the cellular expression of iNOS. Dexamethasone also inhibited GM-CSF-induced NF-kappaB activation by preventing a significant decrease on the IkappaB-alpha protein levels, thus blocking NF-kappaB migration to the nucleus. CONCLUSIONS: The corticosteroid dexamethasone inhibits GM-CSF-induced NF-kappaB activation, iNOS protein expression and NO production. These results suggest that dexamethasone is a potent inhibitor of intracellular events that are involved on NO synthesis, in skin dendritic cells. PMID- 12775355 TI - Anti-inflammatory actions of acupuncture. AB - Acupuncture has a beneficial effect when treating many diseases and painful conditions, and therefore is thought to be useful as a complementary therapy or to replace generally accepted pharmacological intervention. The attributive effect of acupuncture has been investigated in inflammatory diseases, including asthma, rhinitis, inflammatory bowel disease, rheumatoid arthritis, epicondylitis, complex regional pain syndrome type 1 and vasculitis. Large randomised trials demonstrating the immediate and sustained effect of acupuncture are missing. Mechanisms underlying the ascribed immunosuppressive actions of acupuncture are reviewed in this communication. The acupuncture-controlled release of neuropeptides from nerve endings and subsequent vasodilative and anti inflammatory effects through calcitonine gene-related peptide is hypothesised. The complex interactions with substance P, the analgesic contribution of beta endorphin and the balance between cell-specific pro-inflammatory and anti inflammatory cytokines tumour necrosis factor-alpha and interleukin-10 are discussed. PMID- 12775357 TI - Signal transduction pathways in mast cell granule-mediated endothelial cell activation. AB - BACKGROUND: We have previously shown that incubation of human endothelial cells with mast cell granules results in potentiation of lipopolysaccharide-induced production of interleukin-6 and interleukin-8. AIMS: The objective of the present study was to identify candidate molecules and signal transduction pathways involved in the synergy between mast cell granules and lipopolysaccharide on endothelial cell activation. METHODS: Human umbilical vein endothelial cells were incubated with rat mast cell granules in the presence and absence of lipopolysaccharide, and IL-6 production was quantified. The status of c-Jun amino terminal kinase and extracellular signal-regulated kinase 1/2 activation, nuclear factor-kappaB translocation and intracellular calcium levels were determined to identify the mechanism of synergy between mast cell granules and lipopolysaccaride. RESULTS: Mast cell granules induced low levels of interleukin 6 production by endothelial cells, and this effect was markedly enhanced by lipopolysaccharide. The results revealed that both serine proteases and histamine present in mast cell granules were involved in this activation process. Mast cell granules increased intracellular calcium, and activated c-Jun amino-terminal kinase and extracellular signal-regulated kinase 1/2. The combination of lipopolysaccharide and mast cell granules prolonged c-Jun amino-terminal kinase activity beyond the duration of induction by either stimulant alone and was entirely due to active proteases. However, both proteases and histamine contributed to calcium mobilization and extracellular signal-regulated kinase 1/2 activation. The nuclear translocation of nuclear factor-kappaB proteins was of greater magnitude in endothelial cells treated with the combination of mast cell granules and lipopolysaccharide. CONCLUSIONS: Mast cell granule serine proteases and histamine can amplify lipopolysaccharide-induced endothelial cell activation, which involves calcium mobilization, mitogen-activated protein kinase activation and nuclear factor-kappaB translocation. PMID- 12775358 TI - Interleukin (IL)-1 gene polymorphisms: relevance of disease severity associated alleles with IL-1beta and IL-1ra production in multiple sclerosis. AB - BACKGROUND: Multiple sclerosis (MS) is an autoimmune disorder, with a considerable genetic influence on susceptibility and disease course. Cytokines play an important role in MS pathophysiology, and genes encoding various cytokines are logical candidates to assess possible associations with MS susceptibility and disease course. We previously reported an association of a combination of polymorphisms in the interleukin (IL)-1B and IL-1 receptor antagonist (IL-1RN) genes (i.e. IL-1RN allele 2+/IL-1B(+3959)allele 2-) with disease severity in MS. Extending this observation, we investigated whether IL 1beta and IL-1ra production differed depending on carriership of this gene combination. METHODS: Twenty MS patients and 20 controls were selected based upon carriership of the specific combination. In whole blood, in vitro IL-1beta and IL 1ra production was determined by enzyme-linked immunosorbent-assay after 6 and 24 h of stimulation with lipopolysaccharide. RESULTS: Carriers of the specific combination produced more IL-1ra, especially in MS patients, although not significantly. IL-1ra production was significantly higher in individuals homozygous for IL-1RN allele 2. In patients, Il-1ra production was higher and IL 1beta production lower compared with controls. In primary progressive patients, the IL-1beta /IL-1ra ratio was significantly lower than in relapsing-remitting patients. CONCLUSION: Our results suggest higher in vitro IL-1ra production in carriers of IL-1RN allele 2, with an indication of an allelic dose-effect relationship. PMID- 12775359 TI - Allergic diseases, drug adverse reactions and total immunoglobulin E levels in lupus erythematosus patients. AB - BACKGROUND: The association of allergic diseases, drug adverse reactions and elevated total immunoglobulin E (IgE) concentration in systemic lupus erythematosus patients remains controversial. The aim of the study was to investigate the prevalence of those features in active and inactive systemic lupus erythematosus patients, and in the control group as well. METHODS: Total IgE concentration was evaluated by enzyme-linked immunosorbent assay. RESULTS AND CONCLUSIONS: The results of our study revealed that concomitant allergic diseases were not more frequent in systemic lupus erythematosus patients than in the general population. Total IgE concentration was significantly higher during the active stage of the disease. Drug reactions were very frequent but not connected with IgE elevation. Our results indicate that IgE may play a role in lupus pathogenesis, especially in the active phase of the disease. PMID- 12775360 TI - No evidence for a putative involvement of platelet-activating factor in systemic lupus erythematosus without active nephritis. AB - BACKGROUND: Platelet-activating factor (PAF) seems to be implicated in systemic lupus erythematosus (SLE) patients with associated renal diseases. AIMS: In this study, we ensured the role of PAF in SLE patients without renal complications. METHODS: Blood PAF and acetylhydrolase activity, plasma soluble phospholipase A(2), and the presence of antibodies against PAF were investigated in 17 SLE patients without active nephritis and in 17 healthy controls. RESULTS: Blood PAF levels were not different (p=0.45) between SLE patients (6.7+/-2.8 pg/ml) and healthy subjects (9.6+/-3.1 pg/ml). Plasma acetylhydrolase activity (the PAF degrading enzyme) was significantly (p=0.03) elevated in SLE patients (57.8+/-6.4 nmol/min/ml) as compared with controls (37.9+/-2.6 nmol/min/ml). Plasma soluble phospholipase A(2) (the key enzyme for PAF formation) was not different (p=0.6) between SLE patients (59.1+/-5.1 U/ml) and controls (54.7+/-2.4 U/ml). Antibodies against PAF were detected only in 3/17 SLE patients. Flow cytometry analysis did not highlight PAF receptors on circulating leukocytes of SLE patients. CONCLUSION: This clinical study highlights no evidence for a putative important role of PAF in SLE patients without active nephritis. PMID- 12775361 TI - Adenosine deaminase enzyme activity is increased and negatively correlates with catalase, superoxide dismutase and glutathione peroxidase in patients with Behcet's disease: original contributions/clinical and laboratory investigations. AB - AIM: Behcet's disease (BD) is an inflammatory vasculitis with immunologic, endothelial and neutrophil alterations. Adenosine deaminase (AD) is a marker of T cell activation and is related to the production of reactive oxygen species by neutrophils with the production of NO(*), O(2)(*-), H(2)O(2) and OH(*). We reported increased tumour necrosis factor-alpha, soluble interleukin-2 receptor, interleukin-6, interleukin-8 and NO(*) in active BD. As there is a relation between cytokines, T cells and oxidative stress in inflammatory diseases, this study further evaluated: (1) plasma AD activity and its correlation with acute phase reactants; (2) thiobarbituric acid-reactive substances (TBARS) as an indicator for lipid peroxidation; and (3) antioxidant enzymes superoxide dismutase (SOD), glutathione peroxidase (GSHPx) and catalase in patients with BD. The effect of disease activity and correlations between the measured parameters were explored. METHODS: A total of 35 active (n=17) or inactive (n=18) patients with BD (16 men, 19 women) satisfying International Study Group criteria, and 20 age-matched and sex-matched controls (nine men, 11 women) were included in this cross-sectional case-control study. AD and TBARS were measured in plasma, catalase in red blood cells (RBC), and SOD and GSHPx in both plasma and RBC in both groups. Acute phase reactants (alpha(1)-antitrypsin, alpha(2)-macroglobulin, neutrophils, erythrocyte sedimentation rate) were used to classify patients as active or inactive. RESULTS: Plasma AD (mean+/-standard error of the mean, 36.1+/ 0.7 U/l) and TBARS (4.2+/-0.1 nmol/ml) levels were significantly (for each, p<0.001) higher in BD than in controls (24.1+/-0.8 U/l and 1.6+/-0.1 nmol/ml, respectively). RBC catalase activity was significantly (p<0.001) lower in BD than in controls (120.9+/-3.8 versus 160.3+/-4.1 k/g haemoglobin). SOD and GSHPx activities were significantly lower in both plasma and erythrocytes of patients with BD than in controls (plasma SOD, 442.4+/-8.6 versus 636.4+/-9.2 U/ml, p<0.001; RBC SOD, 3719.2+/-66.0 versus 4849.7+/-49.0 U/g haemoglobin, p<0.001; plasma GSHPx, 73.1+/-1.5 versus 90.6+/-2.9 U/ml, p<0.001; RBC GSHPx, 600.7+/-8.0 versus 670.6+/-10.1 U/g haemoglobin, p<0.001). Active BD patients had significantly lower antioxidant enzymes (except RBC catalase) and higher AD and TBARS levels than inactive subjects (for each, p<0.01). When considering all BD patients, a significant positive correlation was present between AD and TBARS (p<0.001) whereas both AD and TBARS were negatively correlated with antioxidant enzymes (for each, p<0.05). CONCLUSIONS: AD and lipid peroxidation are increased and associated with defective antioxidants in BD, suggesting interactions between activated T cells and neutrophil hyperfunction. Measures of pro-oxidative stress and antioxidative defence with AD activity as an indicator of T-cell activation can be considered as significant supportive diagnostic indicators, especially in active disease. In addition, strengthening the antioxidant defence may contribute to treatment modalities. PMID- 12775362 TI - Natural killer cytolytic activity is associated with the expression of killer cell immunoglobulin-like receptors on peripheral lymphocytes in human. AB - Although it has been shown that killer cell immunoglobulin-like receptors (KIRs) on peripheral lymphocytes are upregulated by interleukin-2 (IL-2), which activates natural killer (NK) activity, it has not been demonstrated whether the expression of KIRs is related to NK activity. Therefore, we investigated the association between the KIR expression on lymphocytes and NK activity. CD158a/b expression on lymphocytes obtained from 37 subjects was analyzed using flow cytometry. Simultaneously, NK activity was measured each sample using a 51Cr release assay. Additionally, lymphocytes were cultured in RPMI 1640 medium with or without IL-2 for 48 h, and then their CD158a/b expression and NK activity was analyzed. CD158a/b expression was significantly correlated with NK activity. Especially, the percentage of CD16+CD158a+ and CD8+CD158a/b+ cells in lymphocytes showed a highly significant correlation with NK activity. However, analysis of CD8+ and CD16+ cells revealed that there was only a significant correlation between the percentage of CD8+CD158a+ cells among only CD8+ cells and NK activity. The upregulation of CD16+CD158a+/b+ cells in response to IL-2 tended to be related to the increase of NK activity, but the relationship was not significant. In conclusion, the level of KIR expression was correlated with NK activity, and IL-2 treatment resulted in an increase of NK activity as well as KIR expression, suggesting that upregulation of KIRs enhances the ability to sort target cells, such as virus-infected cells from uninfected cells, according to major histocompatibility complex class I expression. PMID- 12775363 TI - Serum levels of soluble CD30 in adult patients affected by atopic dermatitis and its relation to age, duration of disease and Scoring Atopic Dermatitis index. AB - The value of CD30 and the soluble circulating fragment of CD30 (sCD30) for atopic dermatitis (AD) remains unclear. In particular, little is known about the effects of age, duration of disease and Scoring Atopic Dermatitis index (SCORAD) on the levels of serum sCD30 in patients affected by AD. In the present study, we have analysed serum sCD30 levels of adult patients affected by AD. The study's population includes 18 non-smoking outpatients, with a diagnosis of AD. As a control group we studied 18 non-atopic subjects from laboratory staff, matched for sex and age. These subjects had no history of AD, urticaria or seasonal or perennial rhinitis or asthma, and had negative skin prick test to a panel of allergens. The sCD30 serum levels were clearly higher in patients affected by AD (14.2+/-9.0 IU/ml) than in healthy subjects (1.2+/-0.8 IU/ml) (p<0.001). No differences were observed between males and females affected by atopic dermatitis, regarding age, duration of disease and SCORAD. Significant correlations were found between serum levels of sCD30 levels and age (r=-0.55; 95% confidence interval (CI) for r (Fisher's z transformed)=-0.81 to -0.12; p=0.01), duration of the disease (months) (r=-0.64; 95% CI for r (Fisher's z transformed)=-0.85 to -0.24; p=0.004) and SCORAD (r=-0.74; 95% CI for r (Fisher's z transformed)=-0.89 to -0.42; p=0.004). As demonstrated by the close correlation with age, duration of disease and SCORAD, serum levels of sCD30 appear to be an additional marker for the follow-up of AD. PMID- 12775364 TI - Malondialdehyde in benign prostate hypertrophy: a useful marker? AB - Benign prostate hypertrophy (BPH) is the most common benign tumor in men due to obstruction of the urethra and, finally, uremia. Malondialdehyde (MDA) is a product derived from peroxidation of polyunsaturated fatty acids and related esters. Evaluation of MDA in serum represents a non-invasive biomarker of oxidative stress. Prostate-specific antigen (PSA) is a sensitive marker for prostatic hypertrophy and cancer. We analyzed MDA serum levels to evaluate the oxidative stress in BPH. To this end, 22 BPH patients and 22 healthy donors were enrolled. Data show an increase of MDA level in BPH patients and a positive correlation between PSA and MDA levels. In conclusion, we describe a previously unknown relationship between PSA and MDA as an index of inflammation and oxidative stress in BPH. PMID- 12775365 TI - Safety evaluation of olive phenolic compounds as natural antioxidants. AB - Free and total polyphenolic compounds were extracted from the fruits and leaves of the Picual cultivar. The safety limits of these compounds were recognized by measuring the activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) and the levels of high-density lipoprotein (HDL) cholesterol and total lipids of rat serum. The free and total phenolic compounds (400, 800, and 1600 ppm) and butylated hydroxy toluene (BHT) (200 ppm) were daily ingested for 7 weeks. The administration of olive total and free phenolic compounds at 400 and 800 ppm did not cause any significant changes on ALT and AST activities and serum total lipids. These compounds at 1600 ppm caused significant increase in ALT and AST activities and the content of total lipids. Both olive phenolic compounds were superior to that of BHT in increasing HDL-cholesterol level. Nutritional experiments demonstrated that BHT at 200 ppm caused an enlargement in the kidney and liver of the rat compared with the administration of total and free olive phenolic compounds at 1200 and 1600 ppm. Microscopical examination of kidney and liver tissues of rats administered free and total phenolic compounds at 1200 ppm had the same histological character as that of control rats, while the administration of BHT (200 ppm) and phenolic compounds (1600 ppm) induced severe damage to the tissues of the rat kidney and liver. PMID- 12775366 TI - Selenium composition of cereal-containing infant formulas: assessment of dietary intake status. AB - The selenium (Se) content of 11 infant formulas had been determined using electrothermal and hydride generation atomic absorption spectrometry. The Se content ranged between 0.43 and 17.17 microg/100 g powder infant formulas with wide variations reaching 40.36-fold excess between the poorest RIRI-1 (0.43 microg/100 g) and Milupa-2 (17.2 microg/100 g powder). After reconstituting with water, the adequacies of the 11 studied infant formulas were evaluated with respect to Se supply. Based on the daily adequate intakes (AIs) of 15 and 20 microg Se for children 4-6 and 7-12 months old, respectively, four analyzed infant formulas were found to contain adequate Se content (>75% of the AIs), whereas the other seven formulas could satisfy 63% or less. In conclusion, the national regulation for the trace element (Se) contents in the infant formulas is highly warranted. PMID- 12775367 TI - Nutritional and carbohydrate characteristics of wheat and chickpea based weaning foods. AB - Weaning foods based on malted wheat and chickpea (MWF), popped wheat and chickpea (PWF), and roller dried wheat and chickpea (RWF) were prepared. The PWF and RWF were blended with 5% barley malt flour to formulate low bulk popped (PWFM) and roller dried (RWFM) foods. The amino acid contents, carbohydrate fractionation, pasting characteristics and in vivo carbohydrate digestibility of the foods were determined. Also the liver characteristics of weanling rats fed on the foods were assessed. The foods contained 17.0-18.3% protein, 1.7-3.7% fat, 67.1-67.5% available carbohydrates and 11.1-12.0% dietary fibre. The lysine content of the food proteins ranged from 4.2 to 4.6%. No appreciable difference in the amino acid contents among the different foods and also between the low bulk popped and roller dried foods (PWFM, RWFM) and their high bulk counterparts (PWF and RWF) were observed. The moisture content of the fresh livers of the animals fed on low bulk foods, namely MWF (60.9%), PWFM (63.9%), and RWFM (65.7%), were lower than the high bulk PWF (67.1%) and RWF (70.7%). A similar trend was recorded with respect to fresh and dry weight of the livers. The fractionation of the food carbohydrates by Sepharose CL 2B gel indicated partial hydrolysis of amylopectin fraction and increase in amylose fraction of starch of the MWF, PWFM and RWFM. The PWF also showed a prominent intermediate fraction. The Brabender viscograms of the RWF and PWF exhibited considerable cold paste viscosity, revealing the pregelatinised nature of the starch in these food, but the viscosities of MWF, PWFM and RWFM at 10% slurry concentration were too low to be recorded by the instrument throughout the heating and cooling cycle, probably due to the hydrolysis of their starch by the malt amylases. These observations show that blending about 5% cereal malt with high bulk weaning foods has advantages with respect to reduction in the dietary bulk by partial hydrolysis of carbohydrates, but does not alter their amino acid content significantly. PMID- 12775368 TI - Nutritional quality of nixtamal tortillas fortified with vitamins and soy proteins. AB - The nutritional qualities of nixtamal tortillas (TN) fortified with vitamins and soy flour was evaluated. The rats fed with TN and nixtamal tortillas with 0.15% of vitamins and iron (TNV) did not show differences in weights. The diets of fortified tortillas with 4% defatted soy (TNS), the combination of defatted soy and vitamins (TNVS) and the whole tortilla with defatted soy (TIS) were the treatments that caused the best growth in the experimental animals. The chemical analysis indicated appropriate levels of vitamins in the TNV, TNVS, TNS, and TIS diets. However, the TNV diet caused poor growth of the experimental animals. The diets of tortillas fortified with soy in general produced rats with longer femurs of greater density and strength. The rats fed with fortified tortillas showed higher levels of calcium and phosphorous in the bones. The X-ray diffractograms of bone tissue showed crystals with interplanar distances of 4.40, 3.34, 2.79, 2.28, 1.95, 1.85 and 1.45 A, which are characteristic of hydroxylapatite, except for the first two measurements. The peaks at 4.40 and 3.34 A tend to disappear as the nutritional quality of the diet improves with the addition of soy flour. PMID- 12775369 TI - Temperature effects on fruit quality of mature green tomatoes during controlled atmosphere storage. AB - Tomatoes were harvested at the mature green stage of maturity and stored at 13 degrees C and 15 degrees C for 60 days at three different combinations of CO(2), all with 5.5% O(2) compared with a control with 20.9% O(2) with 0.3% O(2)/% CO(2). Tomatoes were evaluated for changes in colour, titratable acidity, total soluble solids and decay appearance. In this experiment, the 'Criterium' variety of tomatoes was used and it was found that this variety was not suitable for storage at 13 degrees C. Some chilling injury symptoms were observed during the storage period at 13 degrees C, which did not occur at 15 degrees C. Uneven colour development, irregular changes in titratable acidity and total soluble solids were observed during the storage period at 13 degrees C, and the fruits did not ripen properly even after further 10 days ripening time in air at 20 degrees C. Furthermore, the amount of decayed fruits was higher at 13 degrees C than at 15 degrees C, especially after 50 days of storage. In fruits stored for 60 days at 13 degrees C, alterneria rot, blue mould rot and 'cottony leak' occurred that could be associated with chilling injury and increased during a further 10 days ripening at 20 degrees C. It was concluded that the 'Criterium' variety is not suitable for storage at 13 degrees C. PMID- 12775370 TI - Chemical composition of certain tribal pulses in South India. AB - Raw seeds of tribal pulses Atylosia scarabaeoides, Canavalia gladiata, Lablab purpureus var. lignosus, Neonotonia wightii var. coimbatorensis, Rhynchosia filipes, Vigna trilobata and Vigna unguiculata subsp. unguiculata were investigated for their proximate composition, minerals, vitamins (niacin and ascorbic acid) and certain anti-nutritional substances. The seeds of L. purpureus var. lignosus and V. trilobata had a higher content of crude protein than the commonly consumed Indian pulses. The seeds were found to be a rich source of minerals like potassium when compared with recommended dietary allowance values. The total free phenolics, tannins, 3,4-dihydroxyphenylalanine and hydrogen cyanide were also analysed. PMID- 12775371 TI - Investigating Greek consumers' attitudes towards low-fat food products: a segmentation study. AB - The present study aims at gaining a first insight into Greek consumers' attitudes towards low-fat food products. Although Greece, and in particular Crete, have enjoyed a great popularity in terms of the Mediterranean diet, there has been an almost complete lack of low-fat-related surveys concerning the Greek food consumer. Using this as a research trigger, the current investigation evolves around the conflict between 'sensory appeal' and 'healthiness' of low-fat products, widely described in the international literature. Other crucial factors examined are consumers' awareness, occasional use and conscious purchase of, and willingness to pay for, food products with the 'low-fat' claim. Overall, the study has the objective to segment the Greek market in terms of users' perceptions of light products and to identify a number of well-described clusters with clear-cut socio-demographic and behavioural profile. Three clusters are identified, comprised of consumers with favourable attitudes towards low-fat foods and willing to pay premiums to purchase them. PMID- 12775372 TI - Canned rice products as Philippine military food ration. AB - Canned prototypes of rice (CR) and rice meal with pork sausage (CRM) were developed as military food ration models for evaluation by personnel in the Bonifacio Naval Station, Fort Bonifacio, Makati, Philippines. The prototypes were produced based on the assumption that a serving size equivalent to 400 g cooked rice and a meat-based viand using a 1:4 (wt/wt) viand to rice ratio was adequate as a single-serve meal for a typical Philippine military personnel. The CR and CRM prototypes were low acid products with pH values of 4.9 and 5.5, respectively. The processed rice portions of the prototypes showed about 200% volume and weight increases, moderate clumpiness and low percentage breakage. More than 90% of the respondents agreed that CR and CRM were suitable military food rations. Majority of the panelists indicated preference for meat-based and poultry-based viands. Bulk and weight portativity problems, however, were raised with the 400 g serving size of cooked products in cans. PMID- 12775373 TI - Hypoglycaemic and antioxidant effects of onion, Allium cepa: dietary onion addition, antioxidant activity and hypoglycaemic effects on diabetic rats. AB - The purpose of the present study was to discover the relative potency of onion, Allium cepa, with respect to its hypoglycaemic and hypolipidaemic effects on the diabetic situation, and the association of these effects with the potential against oxidative stress. Male Wistar rats were divided into four groups. A normal control (group A), and a non-diabetic group (group B) were treated daily with 1 ml A. cepa solution (0.4 g A. cepa/rat). Groups C and D were made diabetic by an intraperitoneal injection of streptozotocin (STZ) (60 mg/kg body weight) in citrate buffer (pH 6.3). These animals (groups C and D) were the STZ diabetic control and STZ diabetic rats with onion intake, respectively. Onion increased the fasting serum high-density lipoprotein levels, and demonstrated alleviation of hyperglycaemia in STZ diabetic rats. The hypoglycaemic and hypolipidaemic actions of A. cepa were associated with antioxidant activity, since onion decreased superoxide dismutase activities while no increased lipid hydroperoxide and lipoperoxide concentrations were observed in diabetic rats treated with A. cepa. PMID- 12775374 TI - Home hygiene and the prevention of infectious disease in developing countries: a responsibility for all. PMID- 12775375 TI - The infection potential in the home and the role of hygiene: historical and current perspectives. AB - The control of infection through hygiene has a long erratic history. Personal hygiene and handwashing was less appreciated in the past, since it was not known that invisible organisms could spread from apparently clean hands and surfaces. The role of a hygienic home environment received little attention until the 19th century. Since then, the modern tradition of hygiene has served us well, with improved water and drainage developing alongside vaccination, antibiotics, water purification, improved food production and hygienic food preparation and storage. Two major epidemiological trends are relevant to hygiene perspectives: the decline in the morbidity and mortality from infection, and the transition towards higher levels of chronic or debilitating disease. While mortality from some infections has decreased, communicable disease is no less prevalent. Infectious intestinal disease is still unacceptably high in both developed and developing countries. The control of infection within the home needs to take account of changing epidemiological trends, emphasis on evidence-based approaches and loss of public awareness of the role of hygiene. In earlier eras lack of research on the home environment prevented sufficient attention to infection transmission in the domestic setting. Recent research has demonstrated how microbial contamination can be transmitted by activities in the home. Application of this knowledge could significantly reduce the continuing impact of infectious diseases in our communities. PMID- 12775376 TI - Home hygiene and environmental sanitation: a country situation analysis for India. AB - Problems of the environment and of domestic hygiene are always related to poverty of population and the sanitation of settlements. Most cities and towns in developing countries, like India, are characterised by over-crowding, congestion, inadequate water supply and inadequate facilities of disposal of human excreta, waste water and solid wastes. Inadequacy of housing for most urban poor invariably leads to poor home hygiene. Personal and domestic hygiene practices cannot be improved without improving basic amenities, such as water supply, waste water disposal, solid waste management and the problems of human settlements. But even under the prevailing conditions, there is significant scope of improving hygiene practices at home to prevent infection and cross-infection. Unfortunately, in developing countries, public health concerns are usually raised on the institutional setting, such as municipal services, hospitals, environmental sanitation, etc. There is a reluctance to acknowledge the home as a setting of equal importance along with the public institutions in the chain of disease transmission in the community. Managers of home hygiene and community hygiene must act in unison to optimise return from efforts to promote public health. Current practices and perceptions of domestic and personal hygiene in Indian communities, the existing levels of environmental and peri-domestic sanitation and the 'health risk' these pose will be outlined, as well as the need for an integrated action for improving hygiene behaviour and access to safe water and sanitation. PMID- 12775377 TI - Situation analysis and epidemiology of infectious disease transmission: a South East Asian regional perspective. AB - This paper analyses the situation in countries comprising the WHO South-East Asia Region with respect to water supply and sanitation services, hygiene and the epidemiology of related infectious diseases. Recently, published data from the WHO/UNICEF Global Water Supply and Sanitation Assessment 2000 report was reviewed to depict the situation with respect to consumers' access to improved water supply and sanitation services. It was shown that access to improved drinking water supplies is among the lowest in the world, and that sanitation coverage in this region is below all others. The paper also reviews selected surveys of hygiene behaviours in several countries of the region. Associations are suggested between access to services, hygienic practices and specific infectious diseases. The need is acknowledged to improve the evidence base on linkages between infectious diseases and water, sanitation and hygiene, and specific recommendations are made in this regard. There is a need now and for the foreseeable future to promote low-cost household-level interventions, including behaviour change strategies, that mitigate the health consequences of the current situation with respect to water supply, sanitation and hygiene. The role of health authorities in meeting this challenge, and as advocates for accelerating development of the water and sanitation sector, is highlighted. PMID- 12775378 TI - Hygiene and health in developing countries: defining priorities through cost - benefit assessments. AB - Presented here are the four preliminary conclusions in the assessment of health and hygiene in developing countries: (a) child mortality, and disease burden associated with hygiene, water and sanitation in the developing and the developed regions of the world, has declined substantially in the past two decades, but substantial inter-regional and cross-country differences persist; (b) while child mortality and disease burdens decline with higher income levels, a substantial number of countries have been performing far better in reducing child mortality and disease burdens than their income levels would indicate, suggesting that active policy and investment interventions can yield significant health improvements without necessarily jeopardising economic growth; (c) despite the evidence of the role of water and sanitation services in reducing mortality and morbidity, service coverage at the country level has not increased as much as one may have expected in the past decade, in part because of the substantial resource requirements; (d) the paper will provide some new perspectives and evidence on the cost-effectiveness of interventions to reduce the disease burden of poor water and sanitation services and inadequate hygiene practices, in particular with regard to economic evaluation and in reference to hygiene programmes. PMID- 12775379 TI - Epidemiological perspective of domestic and personal hygiene in India. AB - When the application of epidemiology moves from mass phenomenon in a society or community to the specific family or individual level occurrences, new vistas unfold. The classical epidemiological triad, with its multi-mode influences and interactions, becomes modified as a result of several lifestyle factors coming into operation. It is well known that even under severely adverse climatic conditions, microbes are able to survive, and even propagate, if an appropriate micro-climate is encountered. This principle also applies to human beings. Many incidences of disease or ailments, occurrence or absence, can be traced to the home habitat, micro-ecosystem, human behaviour and lifestyles. Hygienic practices are largely a matter of behaviour and usually have biological and social origin. Human behaviour is influenced and determined by social traditions, customs and culture. Furthermore, factors such as health consciousness, practical knowledge of health sciences, motivation and concern for taking steps for promoting health and preventing disease, can change behaviour and make the lifestyle conducive to health. In a village or slum area, families live in more-or-less the same environment. However, in the event of an outbreak of a communicable disease, many escape the attack. While some experience frequent episodes of illness, others continue to live fairly healthily. Obviously, several social and cultural factors and associated human behaviours seem to make the difference between health and disease. This discussion examines the domestic and personal hygiene in its epidemiological perspective. PMID- 12775380 TI - Developing an effective policy for home hygiene: a risk-based approach. AB - Evaluation of the infection potential in the home suggests that improved hygiene practice could significantly reduce the impact of infectious diseases. Fundamental to developing infection prevention policy for the home is the need to recognise that people live in an environment where all human activities occur, including food and water hygiene, hand hygiene, and hygiene related to care of vulnerable groups. In all these situations, reducing infection risks is based on the same underlying microbiological principles. In developing countries, disposal of human and animal excreta and other waste is often also the responsibility of the family and community. Adopting a holistic approach provides the opportunity for a rational approach to home hygiene based on risk assessment. The International Scientific Forum on Home Hygiene (IFH) believes that to deliver hygiene policy with real health benefits, a risk-based approach must be developed and promoted for the home. A risk-based approach starts from the principle that pathogens are introduced continually into homes on people, food and water, pets, insects and air. Inadequate disposal of human and animal excreta serves to increase this risk. Additionally, sites where stagnant water accumulates, such as sinks, toilets and cleaning cloths can support microbial growth and become a source of infection. By assessing the frequency occurrence of pathogens and potential pathogens on hands, hand and food contact surfaces, laundry, reservoir and reservoir/disseminator sites, together with the potential for transfer in the home, the exposure risk can be assessed. PMID- 12775381 TI - Housing improvement projects in Indonesia: responding to local demand. AB - For more than three decades, environmental health programmes in Indonesia have emphasized prevention and treatment of the high incidence of disease among villagers. One of the main causes of disease is the unhygienic conditions of typical rural houses - two-room constructions with dirt floors and walls of lightly fired bricks or woven bamboo skins. While most houses have few or no windows, the occupants frequently cook, eat, sleep and even keep animals in a single room. The main objective of the housing improvement programme was to improve air circulation and introduce more sunlight to kill bacteria, avoid dampness and eliminate smoke from cooking. The programme encourages villagers to construct a permanent floor, enlarge existing windows or insert new windows for good ventilation. This presentation will share the 'success stories' of housing improvement projects in Indonesia that adopted demand-responsive approaches instead of the conventional 'supply approach'. Through exercises like Wealth Classification and Social Mapping, a demand-responsive approach lets the community decide who is eligible for assistance, resulting in higher participation and accurate information on community demand and on materials needed. In addition to the successes, the failures will be discussed at field level. This presentation will discuss the lessons learned from: the World Bank funded Kalisemut Case Study; government's Family Welfare Movement; Plan International's project in Yogyakarta, and AusAID-funded Sustainable Development through Community Participation Project in Lombok. PMID- 12775382 TI - Talking dirty: how to save a million lives. AB - Infectious diseases are still the number one threat to public health in developing countries. Diarrhoeal diseases alone are responsible for the deaths of at least 2 million children yearly - hygiene is paramount to resolving this problem. The function of hygienic behaviour is to prevent the transmission of the agents of infection. The most effective way of stopping infection is to stop faecal material getting into the child's environment by safe disposal of faeces and washing hands with soap once faecal material has contaminated them in the home. A review of the literature on handwashing puts it top in a list of possible interventions to prevent diarrhoea. Handwashing with soap has been calculated to save a million lives. However, few people do wash their hands with soap at these critical times. Obtaining a massive increase in handwashing worldwide requires a sea-change in thinking. Initial results from a new programme led by the World Bank, with many partner organisations, suggest that health is low on people's list of motives, rather, hands are washed to remove dirt, to rinse food off after eating, to make hands look and smell good, and as an act of motherly caring. Professional consumer and market research agencies are being used to work with the soap industry to design professional communications programmes to reach whole populations in Ghana and India. Tools and techniques for marketing handwashing and for measuring the actual impact on behaviour will be applied in new public private handwashing programmes, which are to start up soon in Nepal, China, Peru and Senegal. PMID- 12775383 TI - Handwashing practices and challenges in Bangladesh. AB - Handwashing is universally promoted in health interventions. Studies in Bangladesh and elsewhere have shown a 14 - 40% reduction of diarrhoeal diseases with handwashing. The perceptions and methods related to washing of hands vary widely in Bangladesh. Socio-economic factors are also associated with methods practised. In general, the effectiveness of handwashing practices is poor. Faecal coliform bacteriological counts were reported to be high for both left and right hands. About 85% of women studied who lived in slums and 41% of rural women washed their hands using only water. However, most women rubbed their hands on the ground, or used soil, and rinsed them with water during post-defecation handwashing. Most women claimed that they could not afford to buy soap. Experimental trials showed that use of soap, ash or soil gave similar results when women washed their hands under the same conditions. The washing of both hands, rubbing of hands, and the amount and quality of rinsing water used were found to be important determinants in the reduction of bacterial counts on hands. Although handwashing messages have been revised by most of the main programmes after these studies, there is scope for further improvement, as well as evaluation of their impact. PMID- 12775384 TI - Providing clean water, keeping water clean: an integrated approach. AB - Millions of people, most of whom are children in developing countries, die of basic hygiene-related diseases every year. Interventions in hygiene, sanitation and water supply have been shown to control disease burden. Universal access to improved water sources and basic sanitation remains elusive but is an important long-term goal. Studies have shown that improving the microbiological quality of household water by on-site or point-of-use treatment and safe storage in improved vessels reduces diarrhoeal and other waterborne diseases in communities and households of developing and developed countries. The extent to which improving drinking water quality at the household level reduces diarrhoeal disease probably depends on a variety of technology-related and site-specific environmental and demographic factors that require further investigation, characterisation and analyses. PMID- 12775385 TI - Coping with hygiene in South Africa, a water scarce country. AB - The burden of infectious diseases may be reduced by adopting effective infection control measures. Some of these are dependent on the provision of adequate and safe water supplies for maintenance of basic standards of personal, domestic and healthcare hygiene. Consequences of scarce, and sometimes unsafe, waters supplies in South Africa are highlighted with reference to healthcare-associated infections, community acquired infectious intestinal diseases and domestic practices as infection sources. Availability of water in more than 67% of South African municipal hospitals and primary health care facilities (delivered by water tanker in 12.5% of satellite clinics, 5% from river or dam sources, 12.4% relying on rainwater) does not necessarily guarantee that it's quality is safe for utilisation. In the Northern Province and Mpumalanga, water needs to be purified prior to usage in 14.4 and 33% of satellite clinics respectively. Simple, low maintenance and low-cost interventions to maximise use and safety of limited water resources may be implemented: micro-organism (S. dysenteriae) inactivation by direct UV-exposure in sunlight abundant environments, water purification by filtration mechanisms and making use of iron pots in the community for pasteurisation, decontamination and boiling procedures. Education is paramount in promoting healthy domestic food handling practices, changing cultural perceptions of hygiene, hand-washing technique and mechanisms of domestic environmental decontamination. Water provision cannot be separated from other inter-related factors such as sanitation. Although the present government has taken initiatives to reduce the number of people not having access to water by 50% in 2002, provision of sanitation has been slower (>38% inadequate sanitation in 2002). Adoption of integrated environmental management approaches in conjunction with community participation (WASH Campaign--2002), by the government, aims to address the sanitation problems. PMID- 12775386 TI - Water, sanitation and hygiene: a situation analysis paper for Lao PDR. AB - The Lao People's Democratic Republic (Lao PDR) is located in the Greater Mekong sub-region in East Asia, neighbouring with China, Cambodia, Myanmar, Thailand and Vietnam, with a land area of 236,800 square kilometres, and an average population density of 22 persons per square kilometre. The population of Lao PDR is 5.5 million with 49 ethnic groups. Most of the population is located in rural areas, with a significant number of the neediest villages in remote localities. In the past decade, there have been many positive developments in the rural water supply and sanitation (RWSS) sector. Despite improved coverage in latrine and water supply services, health remains a serious problem. The improved services were often not sustained or poorly maintained, while hygiene received inadequate attention. In Lao PDR, as in many other countries, the provision of a safe and reliable water supply and appropriate sanitation services, based on sustainable approaches, therefore, remains a challenge. This paper will provide an overview of the Lao PDR's RWSS Sector. Special emphasis is placed on actual field level application of informed choices for water supply, sanitation and hygiene awareness. PMID- 12775387 TI - Changing community behaviour: experience from three African countries. AB - In the developing world, more than 1 billion people lack access to safe water. To address this problem, the US Centers for Disease Control and Prevention developed the Safe Water System (SWS), a household-based intervention with three elements: water disinfection, safe storage and behaviour change techniques, and tested these in three countries. In Zambia, social marketing (SM) was used to implement the SWS, and 100 randomly selected households also received motivational interviewing (MI). In Madagascar, the SWS was implemented using SM and community mobilisation (CM). In rural Western Kenya, the SWS was also implemented with SM and CM. In Zambia, 3 months after the SM project launch, 14% of households in the SM-only group had adopted the disinfectant compared with 78% of households in the SM plus MI group. Through SM, over 1 million bottles of disinfectant were sold in 3 years in Zambia. In Antananarivo, Madagascar, 6 months after launch of the water disinfectant, 8% of households in an early stage of the CM process were using the disinfectant compared with 20% in households at a late stage of the CM process. In 1 year, over 500,000 bottles of disinfectant were sold in Madagascar. In Kenya, adoption of the water disinfectant exceeded 60% in intervention households and diarrhoea rates decreased by 58% in children < 5 years. Social marketing permits widespread dissemination of interventions, but may have limited penetration into economically disadvantaged communities. Additional, targeted interventions, such as MI and CM, can increase product adoption. A combination of behaviour change interventions can increase project impact. PMID- 12775388 TI - Sanitation in the developing world: current status and future solutions. AB - More than a third of the world's population (2.4 billion people) lacks access to adequate excreta disposal. Four in five of these unserved people are in Asia, with approximately one in five in both India and China, respectively. Even in large Asian cities, less than half of those served are using sewerage systems; the others use on-site systems, from pit latrines to septic tanks. Most have been installed by householders or builders employed by them, rather than by government or municipal agencies. Governments, international agencies and municipalities can never hope to meet the immense gap in provision unless they promote sanitation with a marketing approach. A latrine is a consumer durable which must be sold. It is often considered that the constraint to increasing sanitation coverage is a lack of demand, but there is often a lack of supply of appropriate products, and latrine designs are often too expensive for the poor, requiring subsidies which are captured by the better-off. More market research is needed to define the right product and how best to stimulate demand. Where subsidies are used, the promotion, not the production of the latrines must be subsidised to prevent middle-class capture of the subsidy. Promotion is probably best performed by different agencies from those that build latrines. The expertise and marketing capacity of the private sector needs to be brought into play, and public bodies must learn to assist it effectively in bringing sanitation to all. PMID- 12775389 TI - Health and social benefits from improving community hygiene and sanitation: an Indian experience. AB - India is a country where Atomic Age and near Stone Age people co-exist. On one hand India has achieved development in many areas, but on the other hand there is still the practice of open defecation and manual cleaning of human excreta from bucket privies by scavengers. National sanitation coverage is only about 34% meaning that 66% of the population practises open defecation. Such unhygienic conditions lead to infections and high mortality and morbidity in the community. Low sanitation coverage could be due to lack of affordable sanitation technology and awareness or motivation. Although the sewerage system was introduced in India long ago, high operational and maintenance costs have prohibited it from being implemented in most towns and cities. Similarly, the cost of a septic tank is beyond most people, and disposal of undigested sludge from septic tanks remains a problem. In contrast, the pour-flush two-pit toilet (known as Sulabh Shauchalaya) is a low cost, socially acceptable and appropriate technology that does not require scavengers to clean the pits. Sulabh has converted and constructed over 1.2 million such toilets throughout India, making 240 towns scavenger-free. Liberated scavengers are thereby available to take up vocational training in various market-oriented trades enabling self-employment. The on-site/ decentralised systems of waste management has improved community health and hygiene, particularly in socially deprived groups, and reduced the financial burden of local government. PMID- 12775390 TI - Myanmar experiences in sanitation and hygiene promotion: lessons learned and future directions. AB - Recent activities in connection with the National Sanitation Week (NSW) and Social Mobilisation for Sanitation and Hygiene have contributed to a significant increase in access to sanitary means of excreta disposal, from 45% in 1997 to 67% in 2001. Handwashing with soap and water after defecation has also increased from 18% in 1996 to 43% in 2001. Success is attributable to high level political commitment, state or division level action and community mobilisation by village level authorities. Multi-level efforts such as mass media, planning workshops, training sessions and house-to-house visits by village authorities and health officials have raised greater awareness of sanitation and hygiene issues and led to construction of latrines on a self-help basis. The challenge ahead is to give greater attention to the 'hard to reach' who live in less accessible areas and are more resistant to change. The 2002 NSW has accordingly given special emphasis to activities in 73 of 324 townships where 50% or more of the households have no access to a sanitary latrine. The communication and social mobilisation package has been improved to upgrading unsanitary latrines and integrating handwashing more systematically with promotion of sanitary latrines. Programmatic follow-up to the NSW is being provided in selected townships through more intensive social mobilisation for 'hard to reach' households and activity-based school sanitation and hygiene education. This approach will contribute further towards improved hygienic practices and reduce diarrhoeal morbidity and mortality. PMID- 12775391 TI - De-worming school children and hygiene intervention. AB - Helminths or worm infestations refer to worms that live as parasites in the human body and are a fundamental cause of disease associated with health and nutrition problems beyond gastrointestinal tract disturbances. Globally, over 3.5 billion people are infected with intestinal worms, of which 1.47 billion are with roundworm, 1.3 billion people with hookworm and 1.05 billion with whipworm. School children aged 5 - 15 years suffer the highest infection rate and worm burden that attributes to poor sanitation and hygiene. About 400 million school age children are infected with roundworm, whipworm and hookworm worldwide, a large proportion of whom are found in the East Asia region (Cambodia, China, Lao PDR, Thailand and Vietnam). These parasites consume nutrients from children they infect, thus retarding their physical development. They destroy tissues and organs, cause abdominal pain, diarrhoea, intestinal obstruction, anaemia, ulcers and other health problems. All of these consequences of infection can slow cognitive development and thus impair learning. De-worming school children by anthelmintic drug treatment is a curative approach for expelling the heavy worm load. However, drug therapy alone is only a short-term measure of reducing worm infection and re-infection is frequent. Control measures through improved sanitation, hygiene and de-worming are needed to prevent infection and re infection. UNICEF has supported many governments in this (and other) regions to assist in the provision of water supply and sanitary facilities and intensive hygiene education in many schools through the Water, Environment and Sanitation (WES) programme. The UNICEF supported school sanitation and hygiene education (SSHE) programme, and other programmes, could effectively enhance behaviour change in children to break the routes of worm transmission and other waterborne diseases. PMID- 12775392 TI - Foodborne diseases in developing countries: aetiology, epidemiology and strategies for prevention. AB - Our knowledge of diseases caused by biologically and chemically contaminated food varies considerably between developing countries. While in Latin America and the Caribbean some information regarding foodborne hazards, high-risk foods and the extent of any resulting disease is available, in many other developing countries little is known about the nature and extent of such diseases. Systematic foodborne surveillance activities, including epidemiological studies, are rarely undertaken. Public health authorities and the public frequently learn about the more dramatic disease outbreaks from news media. A good indication of the importance of food contamination for health and well-being is the information available about infant diarrhoea, infant/child and adult malnutrition. Both conditions are closely related to contaminated food and drinking water, and are particularly important public health problems in most developing countries, as is cholera. With regard to chemicals, little information is available in developing countries on the occurrence of food contamination. Without such information, the health of hundreds of millions of people may be threatened. Improving the safety of the food supply and reducing foodborne diseases requires the concept of shared responsibility to be adopted. All of society needs to accept that food is not only an agricultural and trade commodity but a public health issue. Consequently, Ministries of Health in all countries must integrate food safety as an essential public health function into their work. PMID- 12775393 TI - Effectiveness of home-based food storage training: a community development approach. AB - A community-based home hygiene training initiative, funded by Merseyside Health Action Zone, was piloted in Dingle, a deprived area of Liverpool, UK, during Spring and early Summer 2000. The project actively involved the community, and home-based domestic hygiene training was delivered by community-based facilitators to households in Dingle, Liverpool. During the project the community based facilitators visited 904 households on two separate occasions. During the first visit, the community-based facilitators completed a questionnaire and an 'action sheet' which had been designed to measure participants' prior knowledge and behaviour, in relation to a range of important food hygiene issues, and delivered the home-based food hygiene training. Within 8 weeks of the first visit, the community-based facilitators returned to the participating households and completed a second questionnaire and action sheet. Home-based training is effective in raising awareness, and community development approach has the advantage of facilitating access to households which otherwise would have been difficult to reach. PMID- 12775394 TI - Studies of food hygiene and diarrhoeal disease. AB - Contamination of weaning foods and water with enteropathogenic micro-organisms has been recognised in the past, but its link with the development of diarrhoea by young children in developing countries is lacking. This may explain the unavailability of effective interventions to reduce the risk of diarrhoeal diseases from this contamination. The frequency of contamination of weaning foods with enteropathogens is high in developing countries, and is dependent on the food type, storage time and ambient temperature of storage, the method used, and the temperature reached on re-warming before re-feeding. Other considerations are the bacterial content of cooking and feeding utensils. Fruit and raw vegetables can become contaminated with enteropathogenic micro-organisms by sewage containing irrigation water, by washing produce and fruits in contaminated water, and how they are processed at home. In most studies reviewed, the level of contamination is higher in weaning foods than in drinking water. Since there is a need to reach a critical level of contamination before illness can occur after the ingestion of an enteropathogen, it is postulated that weaning foods are probably more important than drinking water for transmission of diarrhoeal diseases in developing countries. Several potential interventions have been identified, which should be developed and tested in controlled trials in developing countries. These interventions are needed to reduce contamination of weaning foods in households from developing countries, while adequate facilities for the provision of clean water and sanitation to those communities are placed. PMID- 12775395 TI - Moderating effects of psychosocial attributes on the association between risk factors and disability in later life. AB - This study examined the roles of psychosocial attributes in the associations between potential risk factors (age, gender, marital status, education, and chronic conditions) and disability in later life, and in particular how neuroticism and social resources (social network, received support, and satisfaction with support) modify the linkages between risk factors and disability. The main and moderating effects were empirically tested using a sample of 444 community-dwelling older adults in Florida (MU age = 72.3) who were cognitively intact. The likelihood of disability increased with advancing age, more chronic conditions, higher levels of neuroticism, more received support, and less satisfaction w ith support. In addition to the main effects, neuroticism and received support interacted with age and chronic conditions, strengthening the associations between risk factors and disability. Results suggested that personality and social support deserve greater attention as factors that can alter the disability process. PMID- 12775396 TI - Screening for problem drinking in older people referred to a mental health service: a comparison of CAGE and AUDIT. AB - The aim of the study was to evaluate the validity of the Alcohol Use Disorders Identification Test (AUDIT), the five-item version (AUDIT-5) and the CAGE as screening tests for problem drinking in mentally ill older people. The study was of prospective cross-sectional design with questionnaire survey and interview and included all consecutive referrals to an old age psychiatry service fulfilling inclusion criteria. Sensitivity, specificity and positive predictive values and areas under the receiver operating characteristic curves (AUROC) for the AUDIT, AUDIT-5, and CAGE were the primary outcome measures. Using clinical criteria as the gold standard, the AUDIT, AUDIT-5 and CAGE had AUROCs of 0.961, 0.964, and 0.780 respectively. The AUDIT-5 performed best of the three scales with a sensitivity of 75.0%, specificity of 97.2% and positive predictive value of 83.3% when using a 4/5 cut-point. The AUDIT-5 performed as well as the AUDIT and better than the CAGE in identifying problem drinking in this sample. The AUDIT-5 may be a useful addition to the specialist mental health assessment of older people. PMID- 12775397 TI - A confidence interval analysis of three studies using the Alzheimer's disease knowledge test. AB - It has been suggested that community awareness of Alzheimer's disease (AD) has increased over recent years. This claim has been difficult to evaluate given the lack of systematic research in this area however, despite some recent attempts at monitoring changes in knowledge about AD. To address the question of change in awareness about AD, the present study compared results from three studies that have investigated the level of AD knowledge among undergraduate students, using confidence intervals. Consistent with previous findings, the results of this study suggest that more recent cohorts of undergraduate students are more knowledgeable about some aspects of AD, although to an extent that is much less marked than previously thought. Importantly, there are also a number of areas in which there appears to have been little or no change in community awareness about AD, and these are highlighted as issues that may need to be specifically addressed as part of future career education programs. PMID- 12775398 TI - Symptoms of depression in older adults with multiple sclerosis (MS): comparison with a matched sample of younger adults. AB - The study considered for the first time depression in older adults with multiple sclerosis (MS). Depression symptom scores of 27 persons aged over 65 years who were part of a large study of persons with MS (n = 529), were compared with those of a matched sample of younger adults from the same study. The association between cognitive (attitudinal) variables known to explain significant variance in depressive symptoms in younger adults with MS was then considered in the older adult sample. Consistent with findings from studies with general community samples, older adults with MS reported significantly fewer depressive symptoms than younger adults with MS. The relationship between cognitive variables and depressive symptoms found previously in younger adults was also evident for the older adults. Multiple sclerosis related helplessness was found to be significantly higher in older as opposed to younger adults with MS, the opposite of what was predicted given the differences between the groups in depression scores. Differences in the cognitive variables do not appear to explain the differences between older and younger adults with MS in terms of depressive symptoms. This finding offers support for the view that a decrease in emotional responsiveness may explain differences in depressive symptoms between younger and older adults with MS, rather than this being the result of differences in emotional control exerted via cognitive means. PMID- 12775399 TI - A measure of quality of life in early old age: the theory, development and properties of a needs satisfaction model (CASP-19). AB - Quality of life is the subject of much research. However it lacks an agreed theoretical basis. In studies with older populations(ill) health has been used as a proxy measure for quality of life (QoL). We have developed a needs satisfaction measure of QoL in early old age. Our measure has four ontologically grounded domains: conal, autonomy, pleasure, and self-realization. The measure was piloted with focus groups, a self-completion pilot, and cognitive interview testing. This produced a 22-item scale that was included in a postal questionnaire and sent to 286 people aged 65-75 years.A 92% response rate was achieved. The scale was reduced to 19 items on the basis of statistical analysis. The domains have Cronbach's alphas between 0.6 and 0.8. Correlations between the four domains range from 0.4 to 0.7. A second order factor analysis revealed a single latent QoL factor. The scores for the 19-item scale are well distributed along the range although they exhibit a slight negative skew. Concurrent validity was assessed using the Life Satisfaction Index--wellbeing. A strong and positive association was found between the two scales (r= 0.6, p = 0.01). The CASP-19 appears to be a useful scale for measuring QoL in older people. PMID- 12775400 TI - Does standardized needs assessment improve the outcome of psychiatric day hospital care for older people? A randomized controlled trial. AB - There is little evidence to indicate how far psychiatric day hospitals for older people can identify and address the needs of older people who attend. The current practice of Care Programme Approach (CPA) in which multidisciplinary team members, patients and their families meet, may provide a suitable forum for needs to be discussed and interventions planned and implemented. One hundred and twelve new day hospital patients over a period of one year were randomly allocated to an experimental group in which the Camberwell Assessment of Need for the Elderly (CANE). Assessment reports were fed back to day hospital staff but not to the control group in which routine practice was maintained. Similar follow-up assessments were performed after three months in the day hospital or at the time of discharge. At follow-up, both the experimental and control groups had over 60% of their initial unmet needs being adequately met. In both groups, the Health of the Nation Outcome Scales 65+ scores and number of unmet needs were reduced indicating an improvement for both groups. However, there were no significant differences in outcome at follow-up between the experimental and control groups. Day hospitals for older people with mental health problems appear effective in identifying and meeting needs. The feedback from CANE made no difference in terms of outcome. PMID- 12775401 TI - Long-term improvements in cognitive performance through computer-assisted cognitive training: a pilot study in a residential home for older people. AB - The aim of the present pilot study was to investigate the effects of computer assisted cognitive training on aging-associated memory deficits, information processing speed, learning, and interference tendency in older people. Residents of a home for older people (15 women, four men; mean age 83.5; range 75-91) participated in a 14-week computer-assisted cognitive training program. The Niirnberg Aging Inventory and the California Verbal Learning Test were administered prior to the program, immediately after the program and after a period of five months to assess the effectiveness of the cognitive training. After the cognitive training program there were significant improvements in primary working memory and also secondary working memory (for verbal and visual stimuli), on parameters of information processing speed, learning and interference tendency. Improvements in the last two cognitive parameters were maintained five months after completion of the training program. The present study indicates that computerized cognitive training programs can be used in older people to achieve long-term improvements in some important aspects of fluid intelligence. It is suggested that computers could be employed more extensively to prevent and treat cognitive deficits in older people. PMID- 12775402 TI - Coping strategies and its effects on depression among caregivers of impaired elders in Japan. AB - The purpose of this study was to explore the coping strategies of Japanese family caregivers and to investigate relationships between coping strategies, caregiving stressors,and family caregivers' mental health. We conducted a survey by mail among family members who provided care for physically or mentally impaired elderly. In this study, we used 166 responses from family caregivers for analysis. Results of exploratory and confirmatory factor analysis indicated three factors,categorized 'Resignation,' 'Consulting and complaining' and 'Distancing'. In addition, results of structure equation modelling suggested a significant effect of 'Resignation' on the mental health of caregivers. Although the factor structure of coping strategies was similar to other studies conducted in Western countries, the effects of coping strategies were quite different. In conclusion, the effects of coping strategies on caregivers' mental health were confirmed in this study, but different types of coping strategies from those of Western countries were found to be associated with mental health of Japanese family caregivers. PMID- 12775403 TI - Loneliness, health and depression in older males. AB - Loneliness and social isolation, particularly in the older adult, have been shown to influence psychosocial well-being. Loneliness has been related to chronic illness and self-rated health in older adults, and researchers suggest there is an important relationship between loneliness and psychological well-being in older adults particularly in the area of depression. This study investigated relationships between loneliness, health, and depression in 217 older men (> 65 years). Participants completed self-report measures of loneliness, social support, depression, and physical health. Regression analysis showed that a diagnosis of illness or disability was unrelated to depression, however self reported health was associated with depression, with those reporting poorer health experiencing greater depression. Social support variables were unrelated to depression. The most significant relationship to depression was that of loneliness, with lonelier men reporting higher scores on the Geriatric Depression Scale (GDS). Although research suggests that depression is often a response to declining health and functional impairment in the older adult, the present findings suggest that social isolation may also influence the experience of depression. Age-related losses such as loss of professional identity, physical mobility and the inevitable loss of family and friends can affect a person's ability to maintain relationships and independence, which in turn may lead to a higher incidence of depressive symptoms. PMID- 12775404 TI - Baseline experience with Modified Mini Mental State Exam: The Women's Health Initiative Memory Study (WHIMS). AB - The Modified Mini Mental State Exam (3MS) is widely used for screening global cognitive functioning, however little is known about its performance in clinical trials. We report the distribution of 3MS scores among women enrolled in the Women's Health Initiative Memory Study (WHIMS) and describe differences in these scores associated with age, education, and ethnicity. The 3MS exams were administered to 7,480 women aged 65-80 who had volunteered for and were eligible for a clinical trial on postmenopausal hormone therapy. General linear models were used to describe demographic differences among scores. Factor analysis was used to characterize the correlational structure of exam subscales.The distribution of 3MS scores at baseline was compressed in WHIMS compared to population-based data. Mean 3MS scores (overall 95.1) tended to decrease with age and increase with education, however these associations varied among ethnic groups (p< 0.0001) even after adjustment for health, physical disability and occupation attainment. Four factors accounted for 37% of the total variance. Each varied with education and ethnicity; the two most prominent factors also varied with age. Despite relatively narrow distributions in WHIMS, baseline 3MS scores retained associations with age and education. These associations varied among ethnic groups, so that care must be taken in comparing data across populations. PMID- 12775405 TI - Reciprocal relationship between social support and depressive symptoms among Chinese elderly. AB - For many Hong Kong Chinese elderly, depression is a quite common occurrence. This study examines the reciprocal relationship between social support and depressive symptoms. The data came from a longitudinal study of a representative community sample of the elderly population in Hong Kong. Using multiple regression models, the authors found that elderly persons who reported depressive symptoms more frequently were likely to receive higher levels of social support from family members living with elders, but lower levels of social support from friends three years later, even after controlling for socio-demographic and physical health status variables. In addition, elderly people who received more social support from family members not living with elders reported fewer depressive symptoms three years later. PMID- 12775406 TI - Pharmaceutical costs. An overview and analysis of legal and policy responses by the states. PMID- 12775407 TI - Health insurance coverage for oral diseases. Toward a more sophisticated model. PMID- 12775408 TI - Abuse of quarantine authority. The case for a federal approach to infectious disease containment. PMID- 12775409 TI - Covenants not to compete in physician contracts. Recent trends defining reasonableness at common law. PMID- 12775412 TI - Herpes simplex virus latency-associated transcript gene function. AB - A major area of interest in the study of herpes simplex virus type 1 (HSV-1) involves the persistence of the virus within a latent state in neuronal cells of infected humans. The latency-associated transcripts (LATs) are believed to play a key role during HSV-1 latency. This review will discuss the most recent findings on the involvement of the LAT region with apoptotic pathways and how this relates to other potential functions of the LATs. PMID- 12775413 TI - Expression of CCR2, CCR5, and CXCR3 by CD4+ T cells is stable during a 2-year longitudinal study but varies widely between individuals. AB - Blockade of chemokine receptors (CKRs) has recently emerged as a possible pathway for therapeutic intervention in disease. In the present report, the expression of CCR2, CCR5, and CXCR3, associated with migration of mononuclear cells to inflamed tissue, was determined on CD4+ T cells in a 2-year longitudinal study of healthy volunteers using flow cytometry. Large interindividual variations in the expression of these receptors on CD4+ T cells were observed, whereas levels remained remarkably stable over time within subjects. The expression of CCR2, CCR5, and CXCR3 on CD4+ T cells was directly proportional to percentages of CD45RO(hi)/CD4+ T cells. In addition, highly significant associations between levels of CCR2, CCR5, and CXCR3 on CD4+ T cells were demonstrated in individual subjects, implying a common mechanism for regulating the expression of these CKRs on circulating T cells. These associations were not due to coexpression of CKRs on individual CD45RA-/CD4+ T cells. The results provide insight into the regulation of CKR expression on CD4+ T cells in vivo, and suggest that major fluctuations of CKR expression in individuals are uncommon. PMID- 12775415 TI - Adenovirus-mediated expression of CXCL10 in the central nervous system results in T-cell recruitment and limited neuropathology. AB - In the present study, the authors evaluated the contributions of the CXC chemokine ligand (CXCL)10 to leukocyte recruitment into the central nervous system (CNS) and disease development. Instillation of a replication-deficient adenovirus that expresses CXCL10 (AdCXCL10) into the CNS of C57BL/6 mice resulted in a rapid (day 3) and prolonged (day 21) infiltration of both CD4(+) and CD8(+) T cells as compared to mice infected with an adenovirus vector containing beta galactosidase (Adbetagal). Despite increased T-cell infiltration into the CNS of AdCXCL10-infected mice, production of proinflammatory chemokines normally associated with the recruitment of activated T cells into the CNS was muted and mice developed limited neuropathology. Therefore, these results indicate that T cell infiltration in the absence of appropriate activation is not sufficient to induce pathology within the CNS and that additional signals other than CXCL10 are required for induction of an immune-mediated neurologic disease. PMID- 12775414 TI - The chemokine receptor CXCR4 regulates cell-cycle proteins in neurons. AB - Neurons express a variety of chemokine receptors that regulate neuronal signaling and survival, including CXCR4 and CCR5, the two major human immunodeficiency virus (HIV) coreceptors. However, the role of chemokine receptors in HIV neuropathology and neuroinflammatory disorders is still unclear. This study aims to determine whether chemokine receptors regulate the activity of cell-cycle proteins in neurons and evaluate the possibility that alterations of these proteins are involved in HIV neuropathogenesis. The authors studied the effect of the chemokine stromal cell-derived factor (SDF)-1alpha, the natural CXCR4 ligand, and an X4-using variant of gp120 on the activity of cell-cycle proteins involved in neuronal apoptosis and differentiation, such as Rb and E2F-1. Changes in expression, localization, and phosphorylation/activation of Rb and E2F-1 induced by SDF-1alpha (20 nM) gp120(IIIB) (200 pM) were analyzed in primary cultures of rat neurons and in a human cell line expressing recombinant CXCR4. The data indicate that changes in the nuclear and cytosolic levels of Rb--which result in the functional loss of this protein--are associated with apoptosis in hippocampal or cerebellar granule neurons and in cell lines. SDF-1alpha, which is able to rescue these neurons from apoptosis, induces a time-dependent increase of total Rb expression while decreasing the nuclear content of phosphorylated (Ser780/Ser795) Rb and the transcriptional activity of E2F-1. The HIV envelope protein gp120(IIIB) exerts opposite effects at the nuclear level. These data indicate that CXCR4 affects cell-cycle proteins in neurons and raise the possibility that chemokines may contribute to neuronal survival by repressing the activity of E2F-dependent apoptotic genes and maintaining neurons in a highly differentiated and quiescent state. This state may be altered during neuroinflammatory conditions and/or by HIV-derived proteins. PMID- 12775416 TI - Precursors of Borna disease virus-specific T cells in secondary lymphatic tissue of experimentally infected rats. AB - Borna disease in rats represents an experimental model to study the immunopathological role of T cells in central nervous system disease. Adoptive transfer experiments were performed to investigate homing properties of T cells that infiltrate the brains of infected animals. Lymphocytes isolated from the brains of diseased rats were labelled with 5,6-carboxyfluorescein diacetate succinimidyl ester (CFSE) and transferred into immunosuppressed infected recipients. In recipient rats displaying neurological disease, labeled lymphocytes were demonstrated in the vicinity of brain cell lesions, suggesting that the neuronal destruction was dependent on the presence of transferred lymphocytes. Furthermore, the presence of virus-specific cytotoxic T cells was scrutinized in secondary lymphatic tissue and the functional activity of lymphocytes isolated from spleens, cervical lymph nodes, and mesenteric lymph nodes of infected animals was tested immediately after isolation and after in vitro restimulation. The data presented here indicate that precursors of Borna disease virus (BDV)-specific CD8(+) T cells are present and cytotoxic activity was demonstrated after in vitro cocultivation with infected cells in cervical lymph nodes and spleens but not in mesenteric lymphoid tissue. Adoptive transfer of in vitro restimulated T cells induced alterations in BDV-infected, immunosuppressed rats that resemble the well-defined clinical symptoms and neuropathology of Borna disease. This report provides for the first time formal evidence that virus-specific cytotoxic T cells are primed in the periphery after BDV infection, a disease that exclusively manifests itself in the central nervous system. PMID- 12775417 TI - The herpes simplex virus type 1 ICP0 promoter is activated by viral reactivation stimuli in trigeminal ganglia neurons of transgenic mice. AB - Herpes simplex virus type 1 (HSV-1) causes a latent infection in sensory ganglia neurons in humans and in the mouse model. The ability of the virus to latently infect neurons and reactivate is central to the ability of HSV-1 to remain in the human population and spread to new hosts. It is possible that neuronal transcriptional proteins control latency and reactivation by modulating activation of the HSV-1 immediate-early (IE) gene ICP0. We have previously shown that factors in trigeminal ganglia neurons can differentially activate the IE ICP0 promoter and the IE ICP4 promoter in developing trigeminal ganglia neurons of transgenic mice. Ultraviolet (UV) irradiation and hyperthermic stress have been shown to result in HSV-1 reactivation from sensory neurons in the mouse model. Reporter transgenic mice were exposed to UV irradiation or hyperthermia to test whether stimuli that are known to reactivate HSV-1 could activate viral IE promoters in the absence of viral proteins. Measurement of beta-galactosidase activity in trigeminal ganglia from these transgenic mice indicated that the ICP0 promoter activity was significantly increased by both UV irradiation and hyperthermia. The IE genes ICP4 and ICP27 and the late gene gC reporter transgenes failed to be activated in parallel experiments. These results suggest that the ICP0 promoter is a target for activation by host transcription factors in sensory neurons that have undergone damage. It further suggests the possibility that activation of ICP0 gene expression by neuronal transcription factors may be important in reactivation of HSV-1 in neurons. PMID- 12775419 TI - Differential transcriptional regulation by human immunodeficiency virus type 1 and gp120 in human astrocytes. AB - Astrocytes may be infected with the human immunodeficiency virus type 1 (HIV-1) or exposed to the HIV protein gp120, yet their role in the pathogenesis of HIV dementia is largely unknown. To characterize the effects of HIV on astrocytic transcription, microarray analysis and ribonuclease protection assays (RPA) were performed. Infection of astrocytes by HIV or treatment with gp120 had differential and profound effects on gene transcription. Of the 1153 oligonucleotides on the immune-based array, the expression of 108 genes (53 up; 55 down) and 82 genes (32 up; 50 down) were significantly modulated by gp120 and HIV infection respectively. Of the 1153 oligonucleotides on the neuro-based array, 58 genes (25 up; 33 down) and 47 genes (17 up; 30 down) were significantly modulated by gp120 and HIV infection respectively. Chemokine and cytokine induction occurred predominantly by HIV infection, whereas gp120 had no significant effect. These results were confirmed by RPA. The authors conclude that profound alterations of astrocytic function occur in response to HIV infection or interaction with viral proteins, suggesting that astrocytes may play an important role in the pathogenesis of HIV dementia. PMID- 12775418 TI - Human microglial cell isolation from adult autopsy brain: brain pH, regional variation, and infection with human immunodeficiency virus type 1. AB - Microglia are the main source of productive infection by human immunodeficiency virus type 1 (HIV-1) in the central nervous system (CNS). Infection of microglia is difficult to study because nonhuman microglia are not infected by HIV-1, adult human microglia from surgically removed brain tissues are scarce, and fetal human microglial cells differ from adult cells in potentially important ways. Adult autopsies are a potential source of brain tissue for HIV-1 research, but the technique of isolating and infecting cells postmortem is not completely standardized. The authors determined optimal conditions for isolating and infecting adult microglial cells using 18 adult autopsy brain specimens from HIV 1-infected and noninfected decedents. The yield of mixed glial cells overall was on average 0.5 x 10(6) cells per gram of wet tissue. There was no correlation between the number of microglia isolated and the postmortem interval (PMI), HIV seropositivity, age, or gender. Brain pH accounted for about 41% of yield variability; a pH of less than 6.0 generally was not compatible with adequate cell recovery. The highest microglial cell yields were derived from anterior brain sectors (frontal lobe and temporal lobe) versus occipital lobe and cerebellum. A PMI of up to 25.5 h produced excellent cell yields in frontal lobe samples with high brain pH. HIV-1 infection of frontal lobe microglia was 100% successful using both CXCR4- and CCR5-tropic strains of HIV-1. With proper selection of cases and brain region, autopsy brain specimens are a dependable source of viable microglial cells to study CNS HIV-1 infection. PMID- 12775420 TI - Identification of gene products suppressed by human immunodeficiency virus type 1 infection or gp120 exposure of primary human astrocytes by rapid subtraction hybridization. AB - Neurodegeneration and human immunodeficiency virus type 1 (HIV-1)-associated dementia (HAD) are the major disease manifestations of HIV-1 colonization of the central nervous system (CNS). In the brain, HIV-1 replicates in microglial cells and infiltrating macrophages and it persists in a low-productive, noncytolytic state in astrocytes. Astrocytes play critical roles in the maintenance of the brain microenvironment, responses to injury, and in neuronal signal transmission, and disruption of these functions by HIV-1 could contribute to HAD. To better understand the potential effects of HIV-1 on astrocyte biology, the authors investigated changes in gene expression using an efficient and sensitive rapid subtraction hybridization approach, RaSH. Primary human astrocytes were isolated from abortus brain tissue, low-passage cells were infected with HIV-1 or mock infected, and total cellular RNAs were isolated at multiple time points over a period of 1 week. This approach is designed to identify gene products modulated early and late after HIV-1 infection and limits the cloning of genes displaying normal cell-cycle fluctuations in astrocytes. By subtracting temporal cDNAs derived from HIV-1-infected astrocytes from temporal cDNAs made from uninfected cells, 10 genes displaying reduced expression in infected cells, termed astrocyte suppressed genes (ASGs), were identified and their suppression was confirmed by Northern blot hybridization. Both known and novel ASGs, not reported in current DNA databases, that are down-regulated by HIV-1 infection are described. Northern blotting confirms suppression of the same panel of ASGs by treatment of astrocytes with recombinant HIV-1 envelope glycoprotein, gp120. These results extend our previous analysis of astrocyte genes induced or enhanced by HIV-1 infection and together they suggest that HIV-1 and viral proteins have profound effects on astrocyte physiology, which may influence their function in the CNS. PMID- 12775421 TI - Caspase-3-dependent reactivation of latent herpes simplex virus type 1 in sensory neuronal cultures. AB - Life-long latent herpes simplex virus type 1 (HSV-1) is harbored in sensory neurons where sporadic reactivation occurs. Reactivation stimuli may involve activation of apoptotic signaling in the neuron. Previous experiments have demonstrated that reactivation of latent HSV-1 in dorsal root ganglion (DRG) neuronal cultures occurred following nerve growth factor (NGF) deprivation. NGF deprivation stimulates apoptotic signaling by activating the proapoptotic proteolytic enzyme, caspase-3. When DRG neuronal cultures harboring latent HSV-1 were treated with a caspase-3-specific inhibitor, NGF deprivation-induced reactivation was significantly reduced. Interestingly, the caspase-3 inhibitor had no effect on productive HSV-1 infection. Furthermore, activation of caspase-3 with either C2-ceramide or a recombinant adenovirus expressing caspase-3 caused significant HSV-1 reactivation. PMID- 12775422 TI - Human immunodeficiency virus type 1 Tat protein directly activates neuronal N methyl-D-aspartate receptors at an allosteric zinc-sensitive site. AB - The human immunodeficiency virus type 1 (HIV-1) regulatory protein Tat is neurotoxic and may be involved in the neuropathogenesis of HIV-1 dementia, in part via N-methyl-D-aspartate (NMDA) receptor activation. Here, in acutely isolated rat hippocampal neurons, Tat evoked inward currents reversing near 0 mV, with a negative slope conductance region characteristic of NMDA receptor activation. Although the NMDA receptor antagonist ketamine blocked Tat's actions, competitive glutamate- and glycine-binding site antagonists were ineffective (AP 5 and 5,7-dichlorokynurenate, respectively). Evidence for Tat acting at a distinct modulatory site on the NR1 subunit of NMDA receptors was provided by findings that 1 microM Zn(2+) abolished Tat-evoked responses in all neurons tested. Thus, Tat appears to excite neurons via direct activation of the NMDA receptor at an allosteric Zn(2+)-sensitive site. PMID- 12775423 TI - Chronic varicella-zoster virus ganglionitis--a possible cause of postherpetic neuralgia. AB - Postherpetic neuralgia (PHN) is dermatomal distribution pain that persists for months to years after the resolution of herpes zoster rash. The cause of PHN is unknown. Herein, we report clinical, molecular virological, and immunological findings over an 11-year period in an immunocompetent elderly woman with PHN. Initially, blood mononuclear cells (MNCs) contained varicella-zoster virus (VZV) DNA on two consecutive occasions. Random testing after treatment with famciclovir to relieve pain did not detect VZV DNA. However, the patient was reluctant to continue famciclovir indefinitely and voluntarily stopped drug treatment five times. Pain always recurred within 1 week, and blood MNCs contained many, but not all, regions of the VZV genome on all five occasions. Immunological analysis revealed increased cell-mediated immunity to VZV. Chronic VZV ganglionitis induced PHN best explains the recurrence of VZV DNA in MNCs whenever famciclovir was discontinued; the detection of only some regions of the viral genome in MNCs, compared to the detection of all regions of the VZV genome in latently infected ganglia; the increased cell-mediated immunity to VZV; and a gratifying clinical response to famciclovir. The presence of fragments of VZV DNA in MNCs likely represents partial degradation of viral DNA in MNCs that trafficked through ganglia during productive infection. PMID- 12775424 TI - A case of relapsing Guillain-Barre syndrome associated with exacerbation of chronic hepatitis B virus hepatitis. AB - A patient with two episodes of acute polyradiculoneuropathy (Guillain-Barre syndrome) that both occurred during exacerbation of chronic hepatitis B and separated by a 2-year asymptomatic interval is described. The possible causative relation between the neuropathy and the chronic hepatitis B is discussed. PMID- 12775426 TI - The ethics of surgical research. PMID- 12775427 TI - Philip Syng Physick: father of American surgery. PMID- 12775425 TI - Topotecan in the treatment of acquired immunodeficiency syndrome-related progressive multifocal leukoencephalopathy. AB - Progressive multifocal leukoencephalopathy (PML) affects about 1 in 20 individuals with the acquired immunodeficiency syndrome (AIDS) and has been associated with poor survival. This report describes the results of a phase II clinical trial using the drug topotecan, a semisynthetic analogue of camptothecan, administered to a cohort of subjects with AIDS-related PML. Data were evaluated on 11 of 12 subjects enrolled in the study. Three responded to therapy. Additionally, one patient was treated off-protocol and showed a response to treatment. Progression occurred after the first course; however, a partial response was noted after five courses. One study patient died from accidental overdose of topotecan. Overall, responders had higher pretreatment Karnofsky and lower Kurtzke expanded disability status scale scores than nonresponders. The most frequent toxicities were hematologic (anemia, neutropenia, and thrombocytopenia). Five patients had dose delays; all delays were due to hematologic adverse events. This study demonstrates that topotecan treatment may be associated with decreased lesion size and prolonged survival from the infection. Because of the small number of subjects in the study, further studies are required to evaluate the efficacy of topotecan in treating this disease. PMID- 12775429 TI - Role of reactive oxygen species in hepatic ischemia-reperfusion injury and preconditioning. PMID- 12775430 TI - Cytokine cascades and the hepatic inflammatory response to ischemia and reperfusion. AB - Ischemia/reperfusion injury of the liver has two distinct phases that contribute to hepatocyte damage. The acute phase is characterized by Kupffer cell production of reactive oxygen species, which results in moderate hepatocyte injury. The later phase includes an intricate cascade of inflammatory events culminating in neutrophil infiltration of the postischemic liver. Accumulated neutrophils cause substantial injury to hepatocytes through their release of oxidants and proteases. This review discusses the mechanisms by which this inflammatory response is initiated, propagated, and regulated and the impact of these pathways on neutrophil-dependent injury to the liver. PMID- 12775431 TI - Liver ischemia: apoptosis as a central mechanism of injury. AB - Injuries due to cold and warm (normothermic) ischemia remain an important source of morbidity and mortality in liver surgery and transplantation. While the mechanisms of injury are complex, involving apoptotic and necrotic types of cell death, accumulating evidence points to a central role for various apoptotic pathways in mediating cell death following ischemia. A number of antiapoptotic strategies have been shown to confer a high degree of protection, including improved animal survival following long ischemic insult. The future may lie in the identification of practical and effective antiapoptotic strategies to enhance tolerance to ischemic injury. PMID- 12775432 TI - Modified noble plication for the prevention of intestinal intussusception after renal transplantation in dogs. AB - Intestinal intussusception is a frequent problem after experimental transplantation in dogs. This report describes the safety and efficacy of performing a modified Noble plication for the prevention of intussusception. Heterotopic renal transplantation and plication was performed in 20 dogs. Dogs were killed when the serum creatinine concentration exceeded 7 mg/dL because of acute rejection (19 dogs) or venous occlusion (1 dog). Gastrointestinal signs were commonly observed, but no dog experienced an intestinal intussusception, compared to 3 of 14 dogs (21%) previously treated using a similar immunosuppressive regimen. This study supports the routine use of enteroplication in dogs undergoing renal transplantation. PMID- 12775433 TI - Effect of trapidil in ischemia/reperfusion injury on rat small intestine. AB - To investigate the effect of trapidil on the intestinal ischemia-reperfusion injury, we determined malondialdehyde levels as a indicator of lipid peroxidation, nitrite and nitrate levels as reflections of nitric oxide metabolism, and histopathological findings in rats subjected to 40 min of ischemia and 2 h of reperfusion. Histopathological evaluation demonstrated that trapidil treatment has a protective effect on intestinal mucosa and reduces inflammatory cell infiltration in lamina propria, which is consistently noted in the untreated ischemic and reperfused intestines. Possible mechanism of this effect may be explained by the reduced lipid peroxidation (mean malondialdehyde level 3.72 +/- 0.27 vs. 6.13 +/- 0.44, p <.0001) and improved nitric oxide metabolism (mean nitrite plus nitrate 38.21 +/- 2.33 vs. 30.14 +/- 1.47, p =.022). PMID- 12775434 TI - Paracorporeal artificial lung: perioperative management for survival study in sheep. AB - An artificial lung may offer a bridge to recovery or transplant. Utilizing our recently developed paracorporeal artificial lung (PAL) in survival studies in sheep, we critically review our perioperative/anesthetic protocol. Adult Suffolk ewes (n = 15) underwent general anesthesia induced by ketamine (7-15 mg/kg, im) and isoflurane by mask, then intubated and maintained by 4.0-5.0% isoflurane titrated to mean arterial pressure (MAP) 70-110 mm Hg. After a latissimus-sparing thoracotomy and systemic heparinization (200 IU/kg), arterial grafts were anastomosed to the proximal and distal main pulmonary artery in an end-of-graft to side-of-artery fashion. A snare was passed around the pulmonary artery between anastomoses. When the snare was tightened, full pulmonary blood flow was diverted through the cannulae and immediately through the PAL. Perioperative crystalloids included a 500-mL prime, lactated Ringer's (LR) titrated to CVP 5-7 mm Hg, and a heparin infusion (activated clotting time [ACT] 250-300 s). Buprenorphine (0.3 mg im tid) controlled postoperative pain. Hemodynamic parameters, arterial blood gases (ABGs), and ACTs were measured every 6 h. Thirteen of 15 sheep survived the operation and were extubated in less than 20 min. Two groups were studied for up to 7-day survival. Both groups underwent immediate connection to the PAL diverting full pulmonary blood flow. Group 1 (n = 8) underwent immediate connection to a rigidly housed PAL, and 4 of 8 demonstrated immediate right heart failure. In Group 2 a compliance chamber was added to the PAL inflow, and 6 of 7 had stable hemodynamic function for the duration of the study. Incremental improvements in the PAL and our anesthetic and perioperative care have resulted in reliable survival in adult sheep allowing for artificial lung development. PMID- 12775435 TI - New Molecular Approaches for Early Diagnosis and Treatment of Respiratory Diseases. Santa Fe, New Mexico, USA. October 13-16, 2002. Abstracts. PMID- 12775438 TI - Aging and the locus of the global precedence effect: a short review and new empirical data. AB - Precedence of the global over the local level in visual perception is well established by the use of compound, or hierarchical, stimuli. However, (a) virtually nothing is known concerning the fate of global precedence with normal aging, and (b) there is still a debate about the early versus late perceptual locus of the effect. The present study was planned to investigate both topics. Two experiments are reported where young adult and elderly subjects were enrolled for the recognition of compound letters either at the global or at the local level. In order to recruit late perceptual processes, the case of the letters varied in such a way that subjects had to activate graphemic codes to take their decision. It appeared that the global precedence effect was preserved. This observation shows an early perceptual source of the global precedence effect, which is then transferred to later stages of information processing. Age did not affect the global precedence phenomenon. PMID- 12775439 TI - Working memory, language comprehension, and aging: four experiments to understand the deficit. AB - Four experiments were carried out to investigate working-memory capacity and functioning in young and older subjects, and its relation with reading comprehension. Experiment 1 showed that older subjects had to trade off processing and storage functions, due to an assumed reduced pool of resources relative to young subjects. Working-memory measures were weakly correlated with reading comprehension in each age group. Experiment 2 revealed a specific processing impairment in older subjects, and showed that older subjects tended to sacrifice the maintenance of the concurrent mnemonic load to devote more resources to ongoing processing. In Experiment 3, the age-related differences in the processing/storage trade-off were again observed. Furthermore, differences in the speed/accuracy trade-off were revealed. Experiment 4 showed that whatever the instructions regarding the speed/accuracy trade-off, the older subjects remained slower and slightly more accurate than the young subjects. In conclusion, it appears that the age-related differences in working memory are both structural and functional. Different strategies may be implemented in each age group to deal with reading comprehension. PMID- 12775440 TI - Negative associations of chronic stress and cognitive performance in older adult spouse caregivers. AB - Despite research on psychosocial and physical correlates of caregiving, associations of caregiving with cognitive functioning have rarely been studied. We compared the cognitive functioning of 44 spouse caregivers of persons with Alzheimer's disease to 66 demographically-similar non-caregiver spouses; and, whether such differences are explained by demographic variables and distress. We observed that variance in Digit Symbol scores was explained by age [F(1,108)=26.80, p<.001, R(2)=.20] and education [DeltaF(1,107)=5.33, p=.023, DeltaR(2)=.04], and caregiver status [DeltaF(1,106)=4.57, p<.035, DeltaR(2)=.03]. However, when Distress (a composite of Uplifts, Burden, and Sleep Problems) was added to the equation, the lower level of cognitive functioning in caregivers was no longer significant (beta declined from.18 to.06), but distress was significant (beta=-.30). These results may have implications for caregiver adherence and may have relevance to the complex tasks caregivers must perform for care-recipients and the information they report to health care workers/researchers about their care-recipients. PMID- 12775441 TI - Aging affects pointing to unseen targets encoded in an allocentric frame of reference. AB - The goal of the present study was to determine whether aging influences aiming performance to a remembered target encoded in an allocentric frame of reference. We presented four targets simultaneously. The targets were then withdrawn from the screen. Following a recall delay, only three of the targets were presented on the screen in the same configuration as before but at a different location from their first presentation. We asked younger (M=21.9 years) and older participants (M=73.8 years) to point first to the missing target. Because participants did not know where on the computer screen the targets would be presented at the end of the recall delay, they had no choice but to use an allocentric frame of reference to encode its location when the targets were first presented. The results indicate that older participants were significantly more variable when pointing to the remembered target than their younger counterparts, suggesting a decline of allocentric spatial memory with aging. PMID- 12775442 TI - Are changes in cognitive functioning in older adults related to changes in subjective complaints? AB - In many areas of intellectual functioning, age-related declines in older adults' performance and increases in subjective complaints about intellectual performance are observed. However, the literature mostly reports very low relations between functioning and respective complaints. This finding could be the consequence of examining the relation between subjective complaints and the perceived level of functioning. If, however, persons are sensitive to relative changes in performance, they might base their subjective judgment on changes in the level of cognitive functioning. With 202 subjects (mean age=63 years) and two measurements, the authors examine (a) the relation between functioning and complaints at each measurement point, and (b) the relation between changes in functioning and changes in complaints between the measurement points with latent difference variables. The results indicate that there is no relation between level of functioning and level of complaints, but that there is a substantial relation between changes in functioning and complaints. PMID- 12775443 TI - Pictorial illustrations enhance memory for sentences in younger and older adults. AB - In these studies, the authors examined the effects of verbal and pictorial illustrations on younger and older adults' recall of the content of short sentences. During acquisition, base and elaborated sentences were presented under one of three encoding conditions: (a) sentences only; (b) sentences with matching pictures; (c) incomplete sentences with matching pictures. At test, participants recalled the main action of the sentences. Base sentences were recalled more often than elaborated sentences when strict scoring criteria were used. However, older adults showed a recall advantage for elaborated sentences compared to base sentences when lenient scoring criteria were applied. Positive effects of pictorial illustrations occurred for both age groups. Implications for the design of instructional formats to improve older adults' retention of textual material are discussed. PMID- 12775444 TI - Age-associated increases in intensity discrimination for taste. AB - Impaired taste sensation in the aging person may affect the appreciation of food and beverages and compromise nutritional status. Changes in flavor perception may be due to altered ability to discriminate between intensities of suprathreshold taste stimuli. An interesting question is whether all taste qualities (sweet, salty, sour, bitter) show similar age-associated decline in intensity discrimination. Taste intensity discrimination has been shown to be significantly poorer in elderly than in young women for the bitter stimulus caffeine, but not for the sweet stimulus sucrose. The present experiment investigated effects of taste substance and age on taste intensity discrimination by assessing Weber ratios (WRs) for citric acid and sodium chloride (NaCl) in 60 people, half elderly, and half women. Results indicate a significant effect of age on WRs for citric acid and NaCl, suggesting the importance of suprathreshold intensity discrimination for taste perception in the elderly. PMID- 12775446 TI - Effects of mobile phone radiation on UV-induced skin tumourigenesis in ornithine decarboxylase transgenic and non-transgenic mice. AB - PURPOSE: The effects of low-level radiofrequency radiation (RFR) on ultraviolet (UV)-induced skin tumorigenesis were evaluated in ornithine decarboxylase (ODC) and non-transgenic mice. MATERIALS AND METHODS: Transgenic female mice over expressing the human ODC gene and their non-transgenic littermates (20 animals in the cage control group, and 45-49 animals in the other groups) were exposed for 52 weeks to UV radiation or a combination of UV radiation and pulsed RFR. The UV dose was 240 Jm(-2) (1.2 x human minimum erythemal dose) delivered three times a week. One group of animals was exposed to Digital Advanced Mobile Phone System (DAMPS)-type RFR, the other group to Global System for Mobile (GSM)-type RFR at a nominal average specific absorption rate of 0.5 W kg(-1), 1.5 h day(-1), for 5 days a week. The skin was carefully palpated weekly for macroscopic tumours. Histopathological analyses of all skin lesions and of a specified dorsal skin area were performed on all animals. RESULTS: UV exposure resulted in development of macroscopic skin tumours in 11.5 and 36.8% of non-transgenic and transgenic animals, respectively. The RFR exposures did not give a statistically significant effect on the development of skin tumours in either transgenic or non-transgenic animals, or in combined analysis, but tumour development appeared slightly accelerated especially in non-transgenic animals. No effects of RFR exposures were found on excretion of 6-hydroxymelatonin sulphate into urine or on polyamine levels in dorsal skin. CONCLUSION: RFR exposures did not significantly enhance skin tumourigenesis. However, the slightly accelerated tumour development may warrant further evaluation. PMID- 12775447 TI - Low doses of diagnostic energy X-rays protect against neoplastic transformation in vitro. AB - PURPOSE: To investigate the effect of low doses of 60 kVp X-rays on in vitro transformation frequency. MATERIALS AND METHODS: HeLa x skin fibroblast human hybrid cells were used to assay transformation from the non-tumorigenic to the tumorigenic phenotype. Subconfluent cultures of cells were exposed to a range of doses of 60 kVp X-rays and seeded for assay of transformation after 24 h post irradiation holding. Experiments were repeated at least three times and the data pooled for analysis. Transformation frequencies were compared with those of sham irradiated controls. RESULTS: At doses < 1 cGy, the observed transformation frequencies were significantly less than those seen in unirradiated cells. CONCLUSION: Low doses (< 1 cGy) of 60 kVp X-rays protect HeLa x skin fibroblast human hybrid cells against neoplastic transformation in vitro. PMID- 12775448 TI - Relationship between oocyte apoptosis and ovarian tumours induced by high and low LET radiations in mice. AB - PURPOSE: To investigate the biological effectiveness of neutrons at the energy below 1 MeV on apoptosis and carcinogenesis in the mouse ovary. MATERIALS AND METHODS: Female mice were exposed to 1.0 Gy monoenergetic neutrons (0.317, 0.525 and 1.026 MeV), (252)Cf fission neutron (2.13 MeV) or (137)Cs gamma-rays at 7 days of age. Apoptosis of the oocyte and pregranulosa cells, and ovarian carcinogenesis were compared between the radiations. The efficiency of gamma-rays for granulosa cell tumorigenesis was tested by transplantation of the irradiated ovaries into non-irradiated mice. RESULTS: The cumulative apoptotic index of oocytes was 77.9%, 65.6% and 41.6% for the 0.525 MeV neutron, 2.13 MeV neutron and gamma-rays, respectively. Follicles with apoptotic pregranulosa cells were 53.0%, 18.3% and 22.8% of cumulative index for the three groups. Tubular adenomas developed in the groups of monoenergetic neutrons (26.1%) and gamma-ray (35.5%), whereas granulosa cell tumours developed only in the gamma-ray groups (3.2% for 1.0 Gy and 15.6% for 3.0 Gy). Partial-body irradiation with 3 Gy gamma-rays to the ovaries induced granulosa cell tumours with an incidence of 27.3%. CONCLUSION: Effectiveness of neutrons to cause apoptosis was higher for 0.525 MeV than for 2.13 MeV. The pregranulosa cell apoptosis occurred in an oocyte-prone manner. The higher effectiveness of neutrons than gamma-rays to induce oocyte and pregranulosa cell apoptosis correlates with the inhibition of granulosa cell tumour development. PMID- 12775449 TI - Persistence of chromosome aberrations in peripheral lymphocytes from Hodgkin's lymphoma remission patients. AB - PURPOSE: To analyse spontaneous and in vitro bleomycin-induced chromosome aberrations in peripheral lymphocytes taken from Hodgkin's disease patients after prolonged (up to 31 years) remission periods, and to consider these data from the point of view of the carcinogenic potential of anticancer therapy. MATERIALS AND METHODS: Conventional analysis of chromosome preparations stained with azure eosin. RESULTS: The mean frequency and patterns of both spontaneous and induced aberrations in remission patients were significantly different from comparison groups (healthy donors and primary Hodgkin's disease patients). Individual values were characterized with high variation and did not show correlation with post therapy time. New cancer cases diagnosed in remission patients were more frequent in subjects with high chromosome sensitivity to in vitro bleomycin challenge than in patients whose sensitivity to bleomycin was at a control level. CONCLUSIONS: The results are interpreted as suggesting that the tumorigenic potential of radiochemotherapy is mediated via induction of genetic instability in exposed cells. Long after the therapy, the instability may become an initiating event in the development of new malignancies in affected tissues, whereas the instability induced in haemopoietic stem cells may reveal itself in peripheral lymphocytes derived from formerly exposed precursors. PMID- 12775450 TI - Marrow stromal cell recovery after radiation-induced aplasia in mice. AB - PURPOSE: The identification of fibroblast-like cells of the marrow stroma by means of alkaline phosphatase (ALP) cytochemistry reveals delicate ALP-positive structures interspersed among haematopoietic cells and arranged in a loosely meshed network. These cells are often referred to as 'reticular' cells and the network they form is known as the 'ALP network'. The purpose was to analyse the evolution of this ALP network in relation to haemopoietic regeneration after whole-body irradiation. MATERIALS AND METHODS: The total surface occupied by ALP positive processes revealed by means of ALP cytochemistry was expressed as a ratio of the total marrow area. ALP-positive cells were counted using nuclei as the defining unit. Cell proliferation was analysed by the detection of bromodeoxyuridine (BrdU) incorporation. Fat cells were identified by oil red O staining and alpha-glycerophosphate dehydrogenase (alpha-GPDH) activity. RESULTS: The ALP network and ALP-positive cell number began to increase 24 h after 4-Gy irradiation to reach a maximum after 72 h, when the bone marrow was almost completely empty of haemopoietic cells. This increase was in advance of haemopoietic recovery and was not due to cell proliferation. A decrease in the ALP network occurred in parallel with an increase in haemopoiesis and was accompanied by a transient increase in fat cells on day 7. CONCLUSIONS: These data indicate that the recovery of the ALP network, which is partially due to the recruitment of ALP- positive cells, occurs in advance of the haemopoietic recovery and that the equilibrium between fat cells and ALP-positive cells seems to be controlled by haemopoietic cells. PMID- 12775451 TI - C-N bond cleavage reactions on the radiolysis of amino-containing organic compounds and their derivatives in aqueous solutions. AB - PURPOSE: To investigate regularities observed in the course of reactions involving cleavage of C-N bonds on the radiolysis of amino-containing organic compounds and their derivatives in aqueous solutions. MATERIALS AND METHODS: Aqueous solutions of amino-containing compounds, saturated with Ar or O(2), were exposed to gamma-radiation from a (137)Cs source. The absorbed dose-rate was 0.33 Gy s(-1), the absorbed dose range was 0.1-7.5 kGy. Analyses for ammonia and amino containing organic compounds were performed using an amino acid analyser, and analyses for other radiolysis products were made by gas-liquid chromatography. RESULTS: The presence of a hydroxyl group in the beta-position to the amino group promotes deamination of amino alcohols, amino acids, di- and tripeptides, these processes being associated with the possible occurrence of monomolecular fragmentation of the respective radicals. Oxygen inhibits the deamination of alpha,beta-amino alcohols and hydroxyl-containing amino acids. The presence of alkyl substituents in the amino group favours C-N bond cleavage on the radiolysis of amino alcohols, whereas the presence of an acyl group sharply suppresses this process. Replacement of the hydroxyl group in molecules of the initial amino alcohols by a hydrogen atom (i.e. a changeover to alkylamines) or by a methoxy group decreases the efficiency of deamination sharply. The presence of hydroxyl groups in side residues of di- and tripeptides promotes destruction of the peptide chain with formation of amides of amino acids, while suppression of deamination and main chain destruction processes in these compounds is observed in the presence of oxygen. CONCLUSIONS: The most effective processes leading to the C-N bond cleavage on the radiolysis of aqueous solutions of amino-containing organic compounds are those in which the key role is played by reactions of monomolecular decomposition of radicals of the initial compounds. PMID- 12775452 TI - Strong static magnetic field and the induction of mutations through elevated production of reactive oxygen species in Escherichia coli soxR. AB - PURPOSE: Although strong static magnetic fields (SMF) are supposed to have the potential to affect biological systems, the effects have not been evaluated sufficiently. Experiments should be performed with a powerful SMF-generating apparatus to evaluate the biological effects of SMF. MATERIALS AND METHODS: An Escherichia coli mutation assay was used to assess the mutagenic effects of strong SMF. Various mutant strains of E. coli were exposed to up to 9 Tesla (T) for 24 h and the frequencies of rifampicin-resistant mutations were then determined. The expression of the soxS::lacZ fusion gene was assessed by measurement of beta-galactosidase activity. RESULTS: The results for survival or mutation were obtained with wild-type E. coli strain GC4468 and its derivatives defective in DNA repair enzymes or redox-regulating enzymes were all negative. On the other hand, the mutation frequency was significantly increased by the SMF exposure in soxR and sodAsodB mutants, which are defective in defence mechanisms against oxidative stress. Furthermore, the expression of superoxide-inducible soxS::lacZ fusion gene was stimulated 1.4- and 1.8-fold in E. coli when exposed to 5 and 9 T, respectively. CONCLUSIONS: These results indicate that strong SMF induce mutations through elevated production of intracellular superoxide radicals in E. coli. PMID- 12775453 TI - Comments on the paper: modelling radiation-induced chromosome aberrations. PMID- 12775456 TI - Future prospects in medical diagnostics. AB - Medical diagnostics is in the process of redefining its position in everyday medical practice. In future the focus will no longer be merely on measuring pathological parameters in the body but also on their interpretation and their use in the production of guideline-based recommendations for doctors and patients. Interpretations of this kind and 'decision support systems' rely on complex statistical disease models which match individual patients' data with existing evidence-based epidemiological and therapeutic knowledge. The first systems are already available and studies show that in practice they are well accepted by physicians. The following article gives an overview of possible future developments in this field. PMID- 12775455 TI - Electrical impedance tomography (EIT): a review. AB - Electrical impedance tomography (EIT) has been the subject of quite intensive research for about 20 years but has yet to become established as a routine tool in healthcare. None the less the volume of published research work in this area is still rising. This review takes a broad look at what has been achieved and attempts to give the reader sufficient information to form an opinion as to the likely future for this interesting area of research. PMID- 12775458 TI - Laser doppler imaging of menstrual symptoms. AB - In this paper, the skin blood flow for the stomach and forehead regions of 36 female patients with menstrual symptoms was studied using a moorLDI laser Doppler imager in which the results of 6 typical patients are included. The patterns obtained at the two sites are common to all women in the sample who have menstrual symptoms. Cold stress testing was also investigated to see if it was effective in bringing out any skin blood flow fluctuation at these regions caused by menstrual symptoms. Each patient attended two scanning sessions: one before and the other during menstruation. During each session, the patient was scanned three consecutive times, each on the stomach and the forehead skin regions. For each region, the first measurement was a bare scanning whereas for the second and the third, 85% denatured ethanol (cold stress test) was applied onto the required scan areas. It was found that cold stress testing was able to bring out distinct differences in LDI perfusion images before and during menstruation. Results were best captured when perfusion images were taken approximately after 85% denatured ethanol had been applied in two layers for 30 s, allowed to evaporate over the next 5 min (approximately the time taken to obtain one image), reapplied for another 30 s and then finally over the next 30 s allowed to evaporate further. However, it was impossible to deduce conclusively any correlation regarding migraine and skin blood flow since all the patients for this work had menstrual cramps only. PMID- 12775457 TI - On the variability of QRS time-duration in magnetocardiographic recordings. AB - High resolution electrocardiography (HRECG) recordings have already shown an increased beat-to-beat microvariability of the QRS duration of the terminal QRS in patients with a history of ventricular tachycardia (VT). The purpose of this study is to detect QRS-duration microvariability with magnetocardiographic (MCG) recordings in normals, patients with coronary heart disease (CHD), patients with a history of myocardial infarction (MI), and VT patients. QRS microvariability is calculated as the variance of time-shifts of single beats respectively to the average of all beats. The average over all channels of the MCG is performed. QRS microvariability was evaluated from 55-channel MCG in 15 normal persons, in 12 patients with CHD, in 13 patients with MI, and in 10 patients with VT. We found a significantly higher microvariability in patients with MI compared to normals. The highest microvariability was found in VT patients. PMID- 12775459 TI - The non-contact monitoring of heart and respiratory rates using laser irradiation: an experimental simultaneous monitoring with and without clothes during biochemical hazards. AB - The purpose of this study is to develop a non-contact method to evaluate the heart and respiratory rates simultaneously using a single optical sensor which can be used without the removal of clothes before a decontamination procedure in biochemical hazards. We measured the heart and respiratory rates with and without clothes to assess the vital sign monitoring before decontamination. In order to monitor the heart and respiratory rates of rabbits simultaneously, the respiratory and cardiac peaks were separated using fast Fourier transform from a 5 mW helium-neon laser (wavelength 632.8 nm) reflected off the chest walls of rabbits. A cloth (50 mm x 50 mm, 2 mm thick) was placed on the chest of the rabbits to simulate the vital sign monitoring with clothes. The heart rate measured using this method agreed with the rate derived from an electrocardiogram (r = 0.82, p<0.05). The respiratory rate correlated with the manually measured respirator rate (r = 0.93, p<0.05). This method appears promising as a non contact method for monitoring the heart and respiratory rates of patients under biochemically hazardous conditions. PMID- 12775460 TI - Occurrence of C15-C45 mineral paraffins in olives and olive oils. AB - Different classes of olive oils and other olive samples (olives, olive paste and olive pomace) collected during their production were analysed for mineral paraffins in the range of C(15)-C(45). None of the 22 extra virgin olive oils contained mineral paraffins above the detection limit of 1 mg kg(-1). Also, lampante virgin olive oil from the olive mill showed no detectable amounts, but olive oil from the market contained 6-30 mg kg(-1). This contamination cannot be attributed to the refining step, which, on the contrary, partially removes the more volatile hydrocarbons, but could result from transport. Olive-pomace oils obtained by second centrifugation contained 16-145 mg kg(-1) mineral paraffins, presumably because of contamination during storage of the pomace. All olive pomace oils from solvent extraction contained more than 100 mg kg(-1) mineral paraffins, also mainly from storage. Deposition of particulate matter from the air, vehicle exhaust emissions and direct contamination from the bulldozers used to move the pomace were identified as potential sources. PMID- 12775462 TI - Determination of polydimethylsiloxanes by 1H-NMR in wine and edible oils. AB - Fourier transform (1)H-nuclear magnetic resonance (NMR) spectroscopy was suitable for the quantitative determination of polydimethylsiloxanes (PDMS) in wine and edible oil samples. This approach offers highly specific qualitative and quantitative analysis due to silicone-specific location of proton signals linked to carbon atoms located directly next to silicon atoms (0-0.5 ppm), as well as a different location of signals in the range for different organosilicon structures. The method can be used for the control of PDMS at regulatory limits in foodstuffs (10 mg kg(-1)) using hexamethyldisiloxane (HDMS) as an internal standard. Samples were prepared by extraction under suitable conditions to separate the analyte, and with analyte enrichment before (1)H-NMR analysis. Analytical procedures were developed to permit the determination of PDMS at 0.06 mg kg(-1) in wine and at 6 mg kg(-1) in edible oils samples using readily available NMR instrumentation. It was, however, possible to lower the limit of detection to 6 microg kg(-1) for wine and to 60 microg kg(-1) for edible oils using a higher field instrument (500 MHz). Relative standard deviations (S(r)) were obtained for wine (0.028) and for oil samples (0.043), which when compared with values obtained for samples spiked with PDMS (0.021) indicated that the sample preparation was the main factor determining the precision of the method. The average recovery rates for PDMS were 97 and 95% for wine and edible oils, respectively. PDMS was detected in four brands of Italian wine, with Chianti Rafaello containing the highest concentration (0.35 mg kg(-1)), and in four types of edible oils, highest concentration (11.9 mg kg(-1)) being found in Italian corn oil. None of the levels of PDMS found in the food samples exceeded the permissible standards laid down by the Codex Alimentarius Commission (10 mg kg( 1)), with the exception of the one corn oil sample. PMID- 12775461 TI - Differential modulation of enterocyte-like Caco-2 cells after exposure to short chain fatty acids. AB - The response of intestinal epithelial cells to short-chain fatty acids, which are increasingly used as food additives, was investigated. Human small intestinal epithelial cell model Caco-2 cells were exposed to formate, propionate and butyrate to assess their effect on cellular growth, metabolism, differentiation and protection against bacteria. The Caco-2 cells were entirely grown in the different short-chain fatty acids and respective growth patterns were determined. Differentiated cells were exposed to 0-20 mM short-chain fatty acids for 48 h and changes in DNA, RNA, (glyco)protein syntheses, sucrase isomaltase activity, transepithelial electrical resistance and protection against Salmonella enteritidis were measured. The short-chain fatty acids, altered linearly and differentially the growth pattern ranging from stimulation by formate to inhibition by butyrate. Formate inhibited cellular metabolism. Low concentrations of up to 5 mM propionate and 2 mM butyrate stimulated metabolism, while higher doses were inhibitory. Formate had no effect on sucrase isomaltase enzyme activity and transepithelial electrical resistance, whereas propionate and butyrate increased these markers of differentiation. Infection with S. enteritidis did not benefit from the short-chain fatty acid-induced transepithelial electrical resistance. It is concluded that formate, propionate and butyrate selectively and differentially modulate growth characteristics, cellular metabolism, sucrase isomaltase activity and transepithelial electrical resistance in a concentration- and carbon atom-related fashion. The short-chain fatty acid-induced transepithelial electrical resistance does not confer protection against S. enteritidis. PMID- 12775463 TI - Butyltin compounds in retail mollusc products. AB - Butyltin was found in 62 of 74 mollusc products purchased from major supermarket outlets in seven cities across Canada. Tribuyltin was detected in 41 samples, with levels up to 233 ng g(-1). Dibutyltin and monobutyltin levels ranged up to 88 and 53 ng g(-1), respectively. Products originating from East and South East Asia generally contained the highest levels. Gastropods contained lower levels than other molluscs. Enzymatically hydrolysed samples were extracted with 0.05% tropolone in hexane-diethyl ether (1 : 1). Ethyl derivatives were made by Grignard reaction and analysed by gas chromatography coupled to an atomic emission detector. Gas chromatograph-mass spectrometry confirmed the presence of butyltins in the mollusc samples. PMID- 12775464 TI - Cadmium intake of moose hunters in Finland from consumption of moose meat, liver and kidney. AB - Although the average cadmium intake in Finland is about 10 microg day(-1), some risk groups can be identified. This study assessed cadmium intake from the consumption of moose meat, liver and kidneys by moose hunters. Consumption data from a postal questionnaire were combined with a representative database on moose cadmium concentrations. Cadmium intakes were calculated as point estimates for all respondents (n = 711), for those consuming moose meat, liver and/or kidneys, and for the highest decile of those. Probabilistic modelling using the Monte Carlo technique was used to simulate the distribution of dietary cadmium exposure. Of the respondents, 69% consumed moose liver and only 23% moose kidneys. The consumption of moose liver or kidneys significantly increased cadmium intake, whereas moose meat (median consumption 17 kg year(-1) person(-1)) contributed only slightly (0.16 microg day(-1) person(-1)) to the daily total cadmium intake. In the simulation, 10% of the moose hunters had an intake of > 8.76 microg day(-1) (14.6% of PTWI for a 60-kg person) from moose. Point estimates provided only a partial understanding of the potential exposure. Simulated distributions of intake were more useful in characterizing exposure. The study revealed that heavy users of moose organs have a relatively narrow safety margin from the levels of cadmium probably causing adverse health effects. PMID- 12775465 TI - Survey of lead, cadmium, iron, copper and zinc in Kasar cheese. AB - Lead, cadmium, iron, copper and zinc contents of Kasar cheese sold in the markets of Ankara, Turkey, were determined over 12 months. A total of 240 samples comprising 10 different brands were analysed. Graphite-furnace atomic absorption was employed for the determination of lead and cadmium, and flame atomic absorption for iron, copper and zinc. The mean (range) of the lead, cadmium, iron, copper and zinc content of the samples were 86 (10-421) microg kg(-1), 1.8 (0.3-8.3) microg kg(-1), 4.2 (1.0-14.1) mg kg(-1), 0.7 (0.3-1.6) mg kg(-1) and 37.7 (26.5-63.0) mg kg(-1), respectively. The samples in November, December and January contained higher amounts of lead than those in other months (p < 0.01). Moreover, important differences existed in lead content of the samples between different cheese producers (p < 0.01). The differences in cadmium and iron content of the samples for different months were important (p < 0.01). The iron content of the samples among manufacturers also varied significantly (p < 0.01). However, there were no significant differences in copper and zinc contents of the samples over 12 months (p > 0.05). These findings suggested that some contamination occurred during milk production and/or manufacturing of cheese depending on the equipment used. For a consumption of 100 g Kasar cheese, one would ingest approximately 8.6 microg (4% of the provisional tolerable daily intake, PTDI) of lead, 0.2 microg (0.3%) of cadmium, 0.4 mg (0.9%) of iron, 0.07 mg (2%) of copper and 3.8 mg (6%) of zinc. Therefore, it was concluded that Kasar cheese is not a significant contributor to the intake of investigated heavy metals. PMID- 12775466 TI - Aluminium content of Spanish infant formula. AB - Levels of aluminium in 82 different infant formulae from nine different manufacturers in Spain were determined by acid-microwave digestion and graphite furnace atomic absorption spectrophotometry. The influence of aluminium content in tap water in reconstituted powder formulae was examined and an estimate was made of the theoretical toxic aluminium intake in comparison with the provisional tolerable weekly intake (PTWI). Possible interactions between aluminium and certain essential trace elements added to infant formulations have been studied according to the type or main protein-based infant formula. In general, the infant formulae contained a higher aluminium content than that found in human milk, especially in the case of soya, preterm or hydrolysed casein-based formulae. Standard formulae gave lower aluminium intakes amounting to about 4% PTWI. Specialized and preterm formulae resulted in a moderate intake (11-12 and 8 10% PTWI, respectively) and soya formulae contributed the highest intake (15% PTWI). Aluminium exposure from drinking water used for powder formula reconstitution was not considered a potential risk. In accordance with the present state of knowledge about aluminium toxicity, it seems prudent to call for continued efforts to standardize routine quality control and reduce aluminium levels in infant formula as well as to keep the aluminium concentration under 300 microg l(-1) for all infant formulae, most specifically those formulae for premature and low birth neonates. PMID- 12775467 TI - Patulin in domestic and imported apple-based drinks in Belgium: occurrence and exposure assessment. AB - Apple-based beverages are regularly consumed by adults and children in Belgium. They are locally produced or imported from other countries. The apples used as starting material for these productions are frequently contaminated by mycotoxin producing moulds and damaged during transport and handling. The current study was undertaken to investigate whether patulin (PAT) is present in the industrial or handicraft-made apple juices and ciders consumed by the Belgian population and to assess the population's exposure to this mycotoxin through apple-based drinks. Belgian (n = 29) and imported (14) apple juices as well as ciders (7) were assayed for PAT by high-performance liquid chromatography with ultraviolet light detection. PAT was detected respectively in 79, 86 and 43% of these tested samples. However, no contaminated sample exceeded the safe level of 50 microg PAT l(-1). Levels of PAT contamination were comparable in Belgian and imported juice samples. The overall mean PAT concentrations were 9.0 and 3.4 microg l(-1) for contaminated apple juices and ciders, respectively. This study also indicates that there was no statistically significant difference in the mean PAT contamination between clear (7.8 microg l(-1)) and cloudy (10.7 microg l(-1)) apple juices, as well as between handicraft-made apple juices (14.6 microg l(-1)) and industrial ones (7.0 microg l(-1)). On the basis of the mean results, a consumer exposure assessment indicates that a daily intake of 0.2 litres apple juice contributes to 45% of the provisional maximum tolerable daily intake for a child of 10 kg body weight. PMID- 12775468 TI - Naturally occurring phenols: a detoxification strategy for fumonisin B1. AB - Phenolic compounds from plants offer a means for both the prevention and detoxification of mycotoxins that affect human health. This research investigates the control of fungal growth and toxin production by Fusarium verticillioides with plant phenolic compounds, namely chlorophorin, iroko and maakianin, benzoic acid, caffeic acid, ferulic acid, and vanillic acid. Inhibition by these compounds of fungal growth was determined by the agar overlay method and their effect on fumonisin B(1) (FB(1)) production was determined by high-performance liquid chromatography. Chlorophorin was the most effective compound in inhibiting fungal growth, followed by iroko, maakianin, vanillic acid and caffeic acid. Chlorophorin also was the most effective compound in reducing toxin production (94% reduction), followed by caffeic acid, ferulic acid, vanillic acid and iroko, which reduced FB(1) levels by 90-91%. The widespread occurrence of fumonisins world-wide and the lack of adequate prevention of fumonisins require 'biologically safe' alternatives to prevent the transfer of fungi and their health hazardous toxins into our daily foods and environment. PMID- 12775469 TI - Mycotoxins in infant cereal foods from the Canadian retail market. AB - Three hundred and sixty-three samples of cereal-based infant foods were collected from the Canadian retail marketplace over 3 years. The samples included oat-, barley-, soy-, and rice-based infant cereals, mixed-grain infant cereals, teething biscuits, creamed corn, and soy-based formulas. Samples were analysed for targeted mycotoxins (deoxynivalenol, nivalenol, HT-2 toxin, zearalenone, ochratoxin A, fumonisins B(1) and B(2), and five ergot alkaloids). Soy-based cereals (which usually contain corn) exhibited the highest incidences of deoxynivalenol (100%), zearalenone (46%) and fumonisins (75%). Overall, deoxynivalenol was the most frequently detected mycotoxin--it was detected in 63% of samples analysed. Survey results demonstrated the regular occurrence of multiple mycotoxins in cereal-based infant foods. PMID- 12775470 TI - Effects of gamma- and electron-beam irradiation on semi-rigid amorphous polyethylene terephthalate copolymers. AB - Two semi-rigid amorphous polyethylene terephthalate copolymer materials (in both sheet and powder forms) containing 3% 1,4-cyclohexane dimethanol (CHDM) and 31% CHDM were irradiated at 5, 25 and 50 kGy at ambient temperature with a (60)Co radiator or an electron-beam accelerator. After irradiation, volatiles were determined using static headspace sampling with capillary gas chromatography and mass selective detection or flame ionization detection (HS/GC/MSD or FID). Non volatiles were extracted with 10% aqueous ethanol and 100% n-heptane food simulating solvents, maintained at 40 degrees C for up to 10 days. The non volatiles in the materials and those migrating into the food-simulating solvents were determined by high-performance liquid chromatography (HPLC) with ultraviolet and/or photodiode array detection. The results obtained from the HS/GC/MSD suggest that no new chemicals were detected by either gamma- or e-beam irradiation when compared with non-irradiated specimens. The major volatiles in the copolymers were acetaldehyde and 2-methyl-1,3-dioxolane. The concentrations of acetaldehyde increased from 1.24-1.96 mg kg(-1) to 1.94-3.65, 3.52-7.23 and 5.45-15.37 mg kg(-1) after exposure to 5, 25 and 50 kGy doses, respectively. The concentrations of 2-methyl-1,3-dioxolane decreased from 2.49-5.26 mg kg(-1) to 2.07-3.13, 1.33-2.14 and 0.64-2.24 mg kg(-1) after exposure to 5, 25 and 50 kGy doses, respectively. The results of analysis of the copolymers for non-volatiles show that irradiation did not produce any new detectable non-volatile chemicals. A 5 kGy dose had no detectable effect on either copolymer. The 25 and 50 kGy doses had slightly different effects with respect to gamma- and e-beam irradiation on low MW oligomers. However, these increased doses did not significantly affect migration. The concentration of most low molecular weight oligomers migrating into 10% ethanol and 100% heptane was < or =2 ng g(-1) of each oligomer for both copolymers. The cyclic trimer migrating from the 3% CHDM copolymer was approximately 4 ng g(-1); it was 3 ng g(-1) for the 31% CHDM copolymer. The overall results suggest that irradiation significantly increased levels of acetaldehyde but had no effect on non-volatile compounds migrating into food simulants. PMID- 12775472 TI - Estimated daily intake of plasticizers in 1-week duplicate diet samples following regulation of DEHP-containing PVC gloves in Japan. AB - Duplicate hospital diet samples obtained over 1 week in 2001 were analysed to estimate the daily intake of plasticizers and the results were compared with those obtained in 1999. The plasticizers quantified in this study were: dibutyl phthalate, butylbenzyl phthalate (BBP), di(2-ethylhexyl) phthalate (DEHP), diisononyl phthalate (DINP), di(2-ethylhexyl) adipate (DEHA), diisononyl adipate (DINA) and O-acetyl tributyl citrate (ATBC). Dipropyl, dipentyl, dihexyl and dicyclohexyl phthalate were also analysed but not detected. The analytical procedure for this follow-up study was essentially the same as in the previous one. Detection limits were 0.1-15.6 ng g(-1) for each plasticizer. One-week duplicate diet samples provided by three hospitals in three remote prefectures of Japan were analysed as individual meals. DEHP was detected at 6-675 ng g(-1) in 62 of 63 meals, significantly lower levels compared with those detected in 1999. Levels of DEHA and DINP also decreased. The mean intake of plasticizers estimated from all samples was 160 microg DEHP day(-1), 12.5 microg DEHA day(-1), 4.7 microg DINP day(-1) and 3.4 microg BBP day(-1). Levels of DINA were relatively high in meals from one hospital: in those meals, the average daily intake was 1338 microg day(-1). Those of ATBC were also higher in meals from another hospital: the average daily intake was 1228 microg day(-1). The sources of DINA and ATBC can be cling-film or sausage packaging. PMID- 12775473 TI - Simultaneous determination of 4-hydroxy-2-alkenals, lipid peroxidation toxic products. AB - Lipid peroxidation has been associated with cell injury and possible carcinogenecity, for the products can damage cellular membrane and DNA. Among the products formed from lipid peroxidation, 4-hydroxy-2-alkenals are the major components and have been suggested to play a role in cellular toxicity. However, the difficulties of the quantification due to their strong reactivity during laboratory procedures hinder the exposure measurement of the compounds. A novel detection method based on the conversion of 4-hydroxy-2-alkenals to stable diols and to subsequent ortho-esters has been developed. Simultaneous detection in a sensitive single-ion monitoring mode by GC/MS was carried out, monitoring m/z 129, which was the common fragment ion for the ortho-esters derived from 4 hydroxy-2-alkenals. The proposed method was used to quantify 4-hydroxy-2-nonenal in oils and in oxidized arachidonic acid. PMID- 12775475 TI - Analysis of chloramphenicol in honeys of different geographical origin by liquid chromatography coupled to electrospray ionization tandem mass spectrometry. AB - A sensitive liquid chromatography tandem mass spectrometry (LC-MS/MS) method was developed to detect trace amounts of the antibiotic chloramphenicol (CAP) in honey. The methodology entailed a solid-phase extraction of aqueous honey solutions followed by liquid-liquid partitioning, filtration and direct injection onto the LC-MS/MS system. Honey extracts were spiked with an isotopically labelled internal standard (d(5)-CAP) to compensate for analyte loss and potential ion suppression during the MS stage. Detection of the analyte was achieved by negative ionization electrospray in the selected reaction monitoring (SAM) mode. For confirmation, four characteristic mass transitions were monitored each for the analyte and the surrogate standard. The method was validated according to the latest European Union criteria for the analyses of veterinary drug residues in food. At all three fortification levels studied (0.1, 0.2, 0.5 microg kg(-1)) the method was accurate to within 15%. The repeatability and within-laboratory reproducibilities were <12 and 18%, respectively. The decision limit (CC alpha) and detection capability (CC beta) were both <0.1 microg kg(-1). The procedure provides a sensitive and reliable method for the determination of residues of chloramphenicol in honey. Numerous raw honeys of various geographical origins were analysed, showing extensive contamination particularly those of Chinese origin. PMID- 12775474 TI - Response of Vibrio parahaemolyticus 03:K6 to a hot water/cold shock pasteurization process. AB - Vibrio vulnificus and V. parahaemolyticus are natural inhabitants of estuarine environments world wide. Pathogenic strains of these bacteria are often transmitted to humans through consumption of raw oysters, which flourish in the same estuaries. Previous studies reported the effective use of hot water pasteurization followed by cold shock to eliminate from raw oysters naturally and artificially incurred environmental strains of V. vulnificus and V. parahaemolyticus common to the Gulf of Mexico. The present study focused on the use of the same pasteurization method to reduce a highly process resistant Vibrio strain, V. parahaemolyticus O3:K6 to non-detectable levels. Oysters were artificially contaminated with 10(4) and 10(6) V. parahaemolyticus 03:K6 cfu g( 1) oyster meat. Contaminated oysters were pasteurized between 50 and 52 degrees C for up to 22 min. Samples of processed oysters were enumerated for V. parahaemolyticus O3:K6 at 2-min intervals beginning after the 'come-up time' to achieve an oyster internal temperature of at least 50 degrees C. The D value (D(52)deg C) was 1.3-1.6 min. V. parahaemolyticus O3:K6 proved more process resistant than non-pathogenic environmental strains found in Gulf of Mexico waters. A total processing time of at least 22 min at 52 degrees C was recommended to reduce this bacterium to non-detectable levels (< 3 g(-1) oyster meat). PMID- 12775476 TI - Concentrations and bioavailability of cadmium and lead in cocoa powder and related products. AB - Concentrations and bioavailability of cadmium (Cd) and lead (Pb) were determined in cocoa powders and related products (beans, liquor, butter) of different geographical origins. Particular attention was paid to the fractionation of these metals, which was investigated by determining the metal fraction soluble in extractant solutions acting selectively with regard to the different classes of ligands. The targeted classes of Cd and Pb species included: water-soluble compounds, polypeptide and polysaccharide complexes, and compounds soluble in simulated gastrointestinal conditions. The bioavailability of Cd and Pb from cocoa powder, liquor and butter was evaluated using a sequential enzymolysis approach. The data obtained as a function of the geographical origin of the samples indicated strong differences not only in terms of the total Cd and Pb concentrations, but also with regard to the bioavailability of these metals. The Cd concentrations in the cocoa powders varied from 94 to 1833 microg kg(-1), of which 10-50% was potentially bioavailable. The bioavailability of Pb was generally below 10% and the concentrations measured in the cocoa powders were in the 11-769 microg kg(-1) range. Virtually all the Cd and most of Pb were found in the cocoa powder after the pressing of the liquor. PMID- 12775477 TI - From durum wheat to pasta: effect of technological processing on the levels of arsenic, cadmium, lead and nickel--a pilot study. AB - A study was undertaken to investigate the effect of technological processing in pasta-making on the content of arsenic, cadmium, lead and nickel. Milling of durum wheat as well as further processing were carried out in a pilot plant. Commercial pasta samples purchased from the local market were also included for comparison. Furthermore, the effect of cooking was investigated to gain information on the actual content of the selected elements in the final ready-to eat product. Analyte concentrations in whole grain, semolina, pasta and cooked pasta were determined by inductively coupled plasma-mass spectrometry. Milling reduced the element content in the order nickel > arsenic > cadmium > lead. In the experimental conditions of this study, slight contamination by atmospheric lead in milling/pasta-making and release of nickel during pasta-making were observed. These issues have evidently been effectively dealt with in industrial processing given that remarkably lower levels of lead and nickel were found in commercial pastas compared with the experimental samples. On the whole, commercial pasta samples showed low average levels of all the elements included in this study. Cooking caused a significant decrease of the element content in all pasta samples, with average losses of 50-60% on a dry weight basis. PMID- 12775479 TI - Triazole fungicide degradation in peaches in the field and in model systems. AB - The degradation of five triazole fungicides (cyproconazole, hexaconazole, penconazole, propiconazole, tebuconazole) in peaches was evaluated in field studies to establish whether at the preharvest interval the residue levels were below the legal limit established in Italy. All fungicides, except propiconazole, showed higher residues than the legal limits because of their high stability. In fact, some did not decrease at all (cyproconazole, penconazole, tebuconazole), while others decreased only in part. The increasing weight of the fruit during growth was the main factor leading to an apparent decrease in pesticide levels on a weight for weight basis. The trials on model systems showed that co distillation and sunlight photodegradation were the main mechanisms leading to a decrease in triazole levels. Although these fungicides are systemic, they did not enter the fruit and all residues were only present in the peel. PMID- 12775478 TI - Persistent organochlorine pesticides in Mexican butter. AB - Organochlorine pesticides have been used in Mexico in malaria control programmes against ectoparasites and as seed dresser. Owing to their chemical stability, they tend to accumulate in the lipid part of the organisms. The stored pesticides are excreted with the endogenous fat during milk production. The aim was to monitor the organochlorine pesticide levels in butter manufactured in Mexico. From the pesticides, only HCB, beta-HCH, pp'-DDT, op'-DDT and pp'-DDE with major frequency and levels were detected. The HCB mean level was low at 0.008 mg kg(-1) on a fat basis. From the HCH isomer, only the beta-HCH at 0.065 mg kg(-1) on a fat basis was determined, remaining as the main contaminant of the monitored butters. Among DDTs, pp'-DDE was the major constituent (0.043 mg kg(-1) on a fat basis) followed by pp'-DDT (0.036 mg kg(-1)) and op'-DDT (0.009 mg kg(-1)). Comparing the previous study (1994) and this one (2001), all organochlorine pesticides had a descendent tendency; beta-HCH decreased from 0.095 to 0.065 mg kg(-1) on a fat basis, whereas the total DDT decreased from 0.056 to 0.047 mg kg( 1), pp'-DDT from 0.050 to 0.036 mg kg(-1), op'-DDT from 0.018 to 0.009 mg kg(-1), while pp'-DDE increased from 0.032 to 0.043 mg kg(-1). The decreased DDT levels in Mexican butters is caused by the substitution of organochlorine insecticides with pyrethroids used by the Mexican Ministry of Health since 1999 in sanitary programmes. PMID- 12775480 TI - Effects of storage conditions and fruit processing on the degradation of parathion methyl on apples and lemons. AB - Degradation of parathion methyl on lemon and apple on the trees, during storage of these fruits in refrigerated rooms, and during juice production was studied. The pesticide was applied to the trees according to the recommended application procedures. Apples and lemons received a single application of parathion methyl at 40 g active ingredient/100 litres. Residues were determined with a simple gas chromatographic method; the recovery rate of parathion methyl from apples and lemons was 85-108% and the limit of determination was 0.004 mg kg(-1). The half lives of parathion methyl on the trees were 7 days for apples and 5 days for lemons. The duration of storage of the fruits in a refrigerated room have been largely extended to 65 and 63 days, respectively. The high acidity of lemons was found not to affect the degradation rate of parathion methyl. When fruit juice was produced from both apples and lemons, parathion methyl residues were detected in apple but not in lemon juice. PMID- 12775481 TI - Survey of the natural occurrence of zearalenone in maize from northern Iran by thin-layer chromatography densitometry. AB - During September 2000, forty samples of preharvest maize from the province of Mazandaran, north Iran, were randomly collected. Samples were analysed for zearalenone (ZEA) by a thin-layer chromatography (TLC) method (AOAC Official Method). ZEA was extracted with chloroform, purified through a chromatographic column containing silica gel, separated on a TLC plate and quantified by densitometry. The analytical method was validated and was adequately reliable and sensitive. The mean recovery rate of ZEA from spiked samples was 92%. The absolute amount of ZEA standard detectable on a TLC plate was 20 ng, giving a limit of detection (LOD) of 100 ng g(-1). In some samples, it was shown that aflatoxins interfere with ZEA. Therefore, to remove this interference, the TLC mobile phase was changed. Data revealed that three of 40 (7.5%) maize samples contained ZEA in the range 100-212 ng g(-1), with a mean of 141+/-51 ng g(-1). This study, which is the first report of ZEA occurrence in Iranian maize, showed that the ZEA level in maize of Mazandaran province was lower than maximum limit for this mycotoxin in Iran. PMID- 12775482 TI - Improving methods of analysis for mycotoxins: molecularly imprinted polymers for deoxynivalenol and zearalenone. AB - Selective polymeric phases intended for future use in separation/extraction of deoxynivalenol and zearalenone from beverages have been prepared. Using crystalline deoxynivalenol, zearalenone and quercetin, molecularly imprinted polymers were obtained by a non-covalent imprinting approach via a photo initiated addition polymerization. Prepared polymers were based on 4 vinylpyridine, methacrylic acid or 2-trifluoromethylacrylic acid as the functional monomer and on ethyleneglycol dimethacrylate, trimethyltrimethacrylate or divinylbenzene as the cross-linking monomer. Selectivity of the generated molecularly imprinted polymers has been investigated by application of the prepared molecularly imprinted polymers as stationary phases in high-pressure liquid chromatography experiments. The retention and elution behaviours of the template compounds and structurally related substances were determined and compared. The results promise future application of molecularly imprinted polymers as alternative selective matrices for clean-up and enrichment of deoxynivalenol and zearalenone. PMID- 12775483 TI - Deoxynivalenol and other Fusarium toxins in wheat and rye flours on the Danish market. AB - Information on the contamination of Danish cereals and cereal products with Fusarium toxins is limited and the last survey is from 1984/1985. In the present study, the occurrence of deoxynivalenol (DON), nivalenol (NIV), HT-2 toxin, T-2 toxin and zearalenone (ZON) was investigated in flour of common wheat, durum wheat and rye. The samples were collected from 1998 to 2001 from both mills and the retail market in Denmark. A total of 190 flour samples were analysed for DON and NIV and about 60 samples for HT-2, T-2 toxin and ZON. DON was most frequently detected with an incidence rate of 78% over all samples for all years. The contamination level varied considerably from year to year, and for wheat and rye the highest incidence and DON concentrations were found in samples from the 1998 harvest. There were regular and heavy rainfalls in Denmark during the flowering period of the crops that year, and DON was found in all samples, with mean concentrations in wheat and rye flour of 191 microg kg(-1) (n=14) and 99 microg kg(-1) (n=16), respectively. Comparison of data from each harvest year showed higher contents of DON in samples of wheat (range 20-527 microg kg(-1)) than in rye (20-257 microg kg(-1)). Contents of NIV, HT-2 toxin and ZON in samples of wheat and rye were generally low, and even in positive samples the contents were close to the detection limit of the methods. The T-2 toxin was detected in only a few of the wheat samples and in low amounts. However, the toxin was found in about 50% of the rye samples collected during 1998-2000, with a mean content of 49 microg kg(-1) (n=25). Durum wheat flour showed the highest DON contamination level, and all samples (n=33) collected during 2000 and 2001 contained DON with means and medians above 1100 microg kg(-1). Over 70% of the samples contained more than 500 microg kg(-1) DON, and the highest observed concentration was 2591 microg kg(-1). The concentration of T-2 toxin in durum wheat flour was also high with five of the 10 analysed samples containing more than 100 g kg(-1). PMID- 12775484 TI - Occurrence of deoxynivalenol (vomitoxin) in processed cereals and pulses in Turkey. AB - The aim was to investigate the occurrence of deoxynivalenol (DON) in cereal and pulse products in Turkey. DON was detected using high-performance liquid chromatography (HPLC) with ultraviolet detection at 220 nm and positive results greater or equal to 0.60 ppm were confirmed by thin layer chromatography (TLC). An acetonitrile-water (21:4 v/v) extract of the sample was cleaned up on a column packed with alumina-Celite-charcoal (0.35 + 0.25 + 0.40 g). The detection limits for DON were 3 ng/injection (0.10 ppm) and 50 ng/spot (0.60 ppm) for HPLC and TLC, respectively. Eighty-three commercially available cereal and pulse product samples collected from markets and street bazaars were analysed. The recovery rates for boiled, pounded wheat and rice spiked with added DON (1 ppm) were 80.9% (SD 8.37, n=5) and 72.3% (3.85, n=5), respectively. DON was detected in six (8.82%) of 68 cereal and in none of 15 pulse products. The maximum detected amount was 2.67 ppm in a corn flour sample. PMID- 12775485 TI - Development and validation of analytical methods for monomeric and oligomeric migrants from nylon 12 packaging materials. AB - Analytical methods for the determination of laurolactam--the monomer of nylon 12- as well as the cyclic dimer and trimer were established. High performance liquid chromatography using ultraviolet (HPLC-UV) and mass spectrometric detection (HPLC MS) were both found suitable to identify and quantify monomer, cyclic dimer and trimer well below the specific migration limit (SML) of laurolactam, being 5 mg/kg of food (simulant). Gas chromatography with flame ionisation detection (GC FID) showed to be an appropriate method for the detection of only laurolactam in aqueous and fatty food simulants. Food simulants could be analysed directly by all three methods, or after a change of solvents. For olive oil, a method for sample clean-up by size-exclusion chromatography (SEC) was established. PMID- 12775486 TI - Energy cost and physiological responses of males snowshoeing with rotating and fixed toe-cord designs in powdered snow conditions. AB - The purpose of this study was to measure the energy cost and physiological responses of males while snowshoeing with two separate toe-cord designs (rotating toe-cord system vs. fixed toe-cord design) in powdered snow conditions. Eight males snowshoed at self-selected intensity for two, 1600 m trials in two snowshoes, with a rotating toe-cord system and a fixed-toe cord design. It was found that heart rate (HR) (140 vs. 134 beats min(-1)), oxygen consumption (VO(2)) (63.4 vs. 34.0 ml kg(-1)min(-1)), energy cost (56.0 vs. 52.4 kJ min(-1)), and ratings of perceived exertion (RPE) (13 vs. 12) were significantly (p < 0.05) higher while snowshoeing with the fixed toe-cord design than with the rotating toe-cord system. Snowshoeing with the rotating toe-cord system at an average speed of 3.96 km h(-1) produced mean VO(2) values that were 56% of VO(2) max, while snowshoeing with the fixed toe-cord design at 3.86 km h(-1) evoked mean VO(2) values that were 60% of VO(2) max. Mean HR while snowshoeing with the rotating toe-cord system was 70% of HR max, while the mean HR when snowshoeing with the fixed toe-cord design was 74% of HR max. These findings suggest that snowshoeing with a rotating toe-cord system results in lower cardiorespiratory strain in powdered snow conditions compared to snowshoeing with a fixed toe-cord design. PMID- 12775487 TI - Hospital ergonomics: a qualitative study to explore the organizational and cultural factors. AB - The primary objective was to identify the characteristics of the health care industry with respect to organizational and cultural factors and consider how these might impact on the practice of ergonomics. Qualitative methodology was chosen as a suitable approach. This was supported by a middle ground philosophical position. Twenty-one interviews were carried out with academics and practitioners using a questionnaire proforma which developed iteratively over the 18 months of the project. A progressive four stage sampling strategy was used starting with purposive sampling to spread the net. Suggested contacts were then followed up (snowball sampling), before the third stage of intensity sampling to focus on participants with specific experience in hospital ergonomics. A final strategy of analysis sampling sought extreme and deviant cases to achieve theoretical saturation. The analysis resulted in three categories: organizational, staff and patient issues. The organizational issues included both the size and complexity of the National Health Service. For example, three hierarchical lines were identified in the management structure: an administrative line, a professional line and a patient-focused clinical management line. One of the surprising findings for the staff issues was the perceived lack of ergonomic information about female workers as a population group and traditional female employment sectors. The patient issues incorporated three dimensions associated with the caring role: the type of work; expectations; and possible outcomes. The work tends to be dirty and emotional, with a professional subculture to allow the handling of other peoples' bodies. This subculture was linked to a 'coping' attitude where staff put the patients' needs and well-being before their own. The change in patient expectations (from being apologetic through to demanding their rights) is mirrored in a changing model of care from paternalism to partnership. A lack of ergonomic research was identified for female workers in the health care industry relating to both the type of work and gender issues. PMID- 12775488 TI - The effects of job rotation on the risk of reporting low back pain. AB - Job rotation has been widely recommended as an administrative control to reduce the risk of developing work-related musculoskeletal disorders. However, evidence of its benefits are hard to find in the literature. The effect of job rotation on predictions for the risk of reporting low back pain was estimated using Low Back Pain Reporting (LBPR) and Time Weighted Average (TWA) approaches. Index scores calculated using the peak hand force, the peak L4/L5 shear force and the L4/L5 moment cumulated over the entire shift were used to estimate the effects of job rotation on the probability of reporting low back pain. Simulations of realistic rotations between two jobs showed that workers in low demand jobs who rotate into higher demand jobs experience a linear increase in reporting probability using the TWA approach. With the LBPR approach a step increase in reporting probability occurred because of the immediate exposure to the peak loading parameters associated with the more demanding job. With a 50-50 rotation the TWA and LBPR index scores increased by 39% and 57%, respectively. With the LBPR approach the redistribution of risk was not uniform with job rotation. The increase was greater for those who rotated into the demanding job compared to the reduction experienced by those who rotated out of the demanding job. The effects of job rotation are not easily estimated because of the complex effect that mixing jobs has on peak and cumulative tissue loading. PMID- 12775489 TI - Impairments of manual tracking performance during spaceflight are associated with specific effects of microgravity on visuomotor transformations. AB - In contrast to performance in cognitive tasks, tracking performance tends to deteriorate fairly consistently during spaceflight. We address the question whether this decrement results from specific effects of microgravity on motor control or from non-specific effects of the various other stressors present. In a case study we generalize the findings obtained with aiming movements, performed by the same cosmonaut with the same effectors as used for an unstable tracking task, to obtain hypotheses for specific changes of parameters of a simple model used to analyse tracking performance. Consistent with these hypotheses, we observed a reduction of limb stiffness in-flight, but a reduction of the tracking gain post-flight. The cross-task consistency of the observed changes does strongly suggest that the tracking impairment is at least partly caused by specific effects of microgravity on motor control, in particular by a mis calibration of muscular forces which likely results from an underestimation of masses due to weightlessness. PMID- 12775490 TI - Subjective perceptual methods for comparing backpacks in the field. AB - Subjective perceptual methods have provided useful information in the laboratory about small differences in backpack design when physiological and biomechanical comparisons are ineffective, but have never been used in the field. This study therefore evaluated, in a controlled field trial with 10 male participants, the suitability of quantitative and qualitative subjective perceptual approaches to distinguish between subtle design differences in two backpacks, each loaded to 15 kg. In addition, initial quantitative subjective impressions about the two backpacks during a 15 min simulated 'in-shop' trial were compared with post-field trial backpack preference. In the simulated 'in-shop' trial the participants 'tried out' the backpack in a manner that was very similar to the way that they would normally try out a backpack as if they were considering buying one in an 'outdoor' shop. It included donning and doffing the pack several times and walking around the room wearing the backpack. In the controlled field trial, participants carried the two backpacks for approximately 15 min around a 1313 m hilly outdoor track at a self-selected walking pace which elicited a moderate exercise intensity. Seven participants preferred backpack A. Three preferred backpack B. The qualitative approach, which required participants to provide free format written responses to semi-structured open-ended questions immediately after the field trial, successfully identified specific reasons underlying participants' preferences. The main reasons for preferring backpack A were better balance, weight distribution, stability up and down hill and over obstacles, fewer pressure points on their back and easier strap location and adjustment. The quantitative approach, which involved participants responding to written post field trial questions on visual analogue or category ratio rating scales, was generally unsuccessful in distinguishing between backpacks. Thus, qualitative subjective perceptual methods appeared to be more useful than quantitative ones in distinguishing between backpacks and in identifying positive and negative design features under controlled field conditions in which participants carry a backpack at a moderately intense self-selected exercise level. However, since the quantitative approach had been successful in distinguishing between backpacks in an earlier similar study, in which participants exercised more intensely by walking uphill on a treadmill at a fixed pace, it is possible that the quantitative subjective perceptual approach may be capable of distinguishing between backpacks in the field if a fixed pace eliciting higher exercise intensity were to be used. Finally, since quantitative responses to questions about the backpacks after a short simulated 'in-shop' trial closely agreed with participants' post-field trial overall backpack preference, it is concluded that initial subjective impressions may be a good guide to backpack preference after limited field usage. PMID- 12775491 TI - The effects of posture on forearm muscle loading during gripping. AB - The purpose of this study was to quantify the response of the forearm musculature to combinations of wrist and forearm posture and grip force. Ten healthy individuals performed five relative handgrip efforts (5%, 50%, 70% and 100% of maximum, and 50 N) for combinations of three wrist postures (flexed, neutral and extended) and three forearm postures (pronated, neutral and supinated). 'Baseline' extensor muscle activity (associated with holding the dynamometer without exerting grip force) was greatest with the forearm pronated and the wrist extended, while flexor activity was largest in supination when the wrist was flexed. Extensor activity was generally larger than that of flexors during low to mid-range target force levels, and was always greater when the forearm was pronated. Flexor activation only exceeded the extensor activation at the 70% and 100% target force levels in some postures. A flexed wrist reduced maximum grip force by 40-50%, but EMG amplitude remained elevated. Women produced 60-65% of the grip strength of men, and required 5-10% more of both relative force and extensor activation to produce a 50 N grip. However, this appeared to be due to strength rather than gender. Forearm rotation affected grip force generation only when the wrist was flexed, with force decreasing from supination to pronation (p < 0.005). The levels of extensor activation observed, especially during baseline and low level grip exertions, suggest a possible contributing mechanism to the development of lateral forearm muscle pain in the workplace. PMID- 12775494 TI - Genomics and the search for novel biomarkers in toxicology. AB - The advent of 'genomics' technology, in particular transcript profiling, has already had a measurable impact on the drug discovery process in the areas of target identification and validation. This review is concerned with the potential application of this technology to toxicology and drug safety assessment, with particular emphasis on biomarker discovery and characterization. An advantage (or possibly a drawback!) of transcript profiling is that candidate biomarkers of toxicity can be speedily identified, with the caveat that a significant amount of subsequent experimental and bioinformatic effort needs to be expended in order to evaluate and validate them. Attention is also drawn to the critical need for robust experimental design with studies of this type and to issues associated with the analysis of large data sets. In summary, while genomics technology undoubtedly offers much that can assist drug safety assessment, its potential has yet to be realized fully in this area. However, a large amount of resource continues to be applied to 'toxicogenomics'. Tangible benefits, in terms of new biomarkers of toxicity and reduced numbers of adverse drug effects, remain realistic objectives. PMID- 12775495 TI - Simultaneous determination of urinary 1- and 2-naphthols, 3- and 9-phenanthrols, and 1-pyrenol in coke oven workers. AB - A method was developed for simultaneous quantification of urinary 1- and 2 naphthols, 3- and 9-phenanthrols and 1-pyrenol using gas chromatography with mass spectrometry (GC-MS). This method was applied to urine samples from coke oven workers (n=28) and controls (n=22) from Northern China. Geometric mean levels of urinary 1-naphthol (58.8 microg l(-1)), 2-naphthol (34.1 microg l(-1)), 3 phenanthrol (7.35 microg l(-1)), 9-phenanthrol (1.28 microg l(-1)) and 1-pyrenol (25.4 microg l(-1)) were significantly higher among coke oven workers than controls. All the substances tested were highest among top-of-oven workers, who had 15-fold higher 1-naphthol, eight-fold higher 2-naphthol and 20-fold higher 1 pyrenol levels compared with controls. Using multiple linear regression models, 72.5% of the variation in 1- and 2-naphthol and 82.8% of the variation in 1 pyrenol were explained by the concentration of naphthalene or pyrene in the urine, the work category and the smoking intensity. Cigarette consumption significantly contributed to levels of urinary 1-pyrenol and naphthols, particularly 2-naphthol. A negative relationship between work category and the ratio of naphthols/1-pyrenol was observed among smokers. Our results suggest that urinary naphthols and phenanthrols reflect polycyclic aromatic hydrocarbon (PAH) exposure as well as the widely used 1-pyrenol, and that interactions between cigarette smoking and PAH exposure result in different patterns of metabolism for individual PAHs. PMID- 12775496 TI - Fumonisin mycotoxins in human hair. AB - This study shows for the first time the accumulation of fumonisin mycotoxins in human hair of population clusters exposed to contaminated maize, and thus the feasibility of human hair analysis for the assessment of past fumonisin exposure. Composite hair samples were obtained from the Bizana, Butterworth and Centane districts within the Transkei region of the Eastern Cape Province of South Africa. Following methanol extraction and strong anion exchange clean up, the fumonisins FB(1), FB(2) and FB(3) were detected using high performance liquid chromatography coupled to electrospray ionization-mass spectrometry (HPLC-ESI MS). Hair from Centane and Butterworth showed mean levels of FB(1) of 26.7 and 23.5 microg kg(-1) hair, respectively. FB(2) was only detected in hair from Centane and in one sampling point in Butterworth, with mean levels of 6.5 and 5.7 microg kg(-1) hair, respectively. Hair samples from Bizana, on the other hand, were found to contain higher levels of FB(1) (mean 33.0 microg kg(-1) hair) and FB(2) (mean 11.1 microg kg(-1) hair). No samples contained more than trace levels of FB(3). Recoveries from spiked hair samples using this method ranged from 81% to 101%, demonstrating the applicability of hair analysis in assessing human exposure to fumonisin mycotoxins. PMID- 12775497 TI - Determination of urinary hippuric acid and o-cresol levels as biological indicators of toluene exposure in shoe-workers and glue sniffers. AB - In this study, groups exposed to toluene either intentionally (glue sniffers) or unintentionally (shoe-workers) were compared. The groups were evaluated in terms of urinary levels of the toluene metabolites hippuric acid and o-cresol. Results were also compared with control values. Hippuric acid levels were determined by high performance liquid chromatography and o-cresol levels by gas liquid chromatography. The levels of hippuric acid and o-cresol were found to be statistically significantly higher in glue sniffers than in shoe-workers (p <0.001) or controls (p <0.001). In addition, the differences between the levels of urinary hippuric acid and o-cresol in the shoe-workers and in the controls were statistically significant (p <0.05 and p <0.001, respectively). These results suggest that extremely high levels of urinary hippuric acid and o-cresol indicate massive exposure to toluene. PMID- 12775498 TI - Metallothionein induction related to hepatic structural perturbations and antioxidative defences in roach (Rutilus rutilus) exposed to the fungicide procymidone. AB - A variety of stresses, hormones, glucocorticoids and cytokines are known to induce metallothioneins (MTs) in animals. The aim of this study was to investigate the effects of chemical stress induced by the dicarboximide fungicide procymidone on hepatic structure, MT content and antioxidative defences (catalase and glutathione reductase activities and glutathione content) in the common fish Rutilus rutilus. Catalase and glutathione reductase activities remained stable throughout the experiment. Four days of exposure to 0.2 or 0.4 mg l(-1) of procymidone induced an obvious increase in liver MT content, perturbation of metal MT contents, and an increase in hepatic glutathione content. After 14 days' exposure, obvious and large structural alterations of the hepatic parenchyma occurred simultaneously with a decrease in MT and glutathione content. These events were interpreted as degeneration of the liver. Fish exposed for 14 days to procymidone and then placed for 14 days in clean water showed nearly complete decontamination of the liver, but MT concentrations remained high. The toxicological significance of these events is discussed. PMID- 12775499 TI - A cross-sectional study of polycyclic aromatic hydrocarbon-DNA adducts and polymorphism of glutathione S-transferases among heavy smokers by race/ethnicity. AB - Differences in lung cancer risk by race/ethnicity have been observed among smokers. To determine whether these observations might reflect differences in the formation of carcinogen-DNA adducts, we analysed blood specimens (n=151) collected from smokers who were recruited for possible participation in an antioxidant vitamin intervention study. Mononuclear cells were analysed for polycyclic aromatic hydrocarbon (PAH)-DNA adducts by competitive enzyme-linked immunosorbent assay. Genotypes of glutathione S-transferase M1 and P1 (GSTM1 and GSTP1), enzymes involved in the detoxification of PAH metabolites, were determined by polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism, respectively. GSTM1 was present in 65 out of 88 (73.4%), 16 out of 32 (50.0%) and 16 out of 29 (54.8%) of African-Americans, Caucasians and Latinos, respectively (p=0.022). Homozygosity for the GSTP1 codon 105 variant was found in 25.6%, 6.3% and 10.0% of African-Americans, Caucasians and Latinos, respectively (p=0.023). Regression analysis of the log-transformed adduct levels confirmed that Caucasian and Latino subjects had lower PAH-DNA adduct levels than African American subjects, after adjustment for gender, education, alpha-tocopherol and beta-carotene levels, and GSTM1 status. Further adjustment for age and current smoking habits had no impact on these findings. Although crude analysis suggested that the GSTM1-positive genotype may be associated with lower PAH-DNA levels in Caucasians (but not in African-Americans or Latinos), a formal test for interaction between GSTM1 and ethnicity was not significant. We found no association between adduct levels and GSTP1 genotype. Although the mechanism is unclear, ethnic differences in DNA damage levels may in part explain why African Americans have higher lung cancer incidence rates than other ethnic groups. PMID- 12775500 TI - Comparison of K-ras gene mutations in tumour and sputum DNA of patients with lung cancer. AB - Mutations in the K-ras gene are frequently found in lung tumours and are implicated in the development of lung cancer. In order to investigate the clinical usefulness of these mutations in lung cancer, we applied a sensitive method to compare mutations in codon 12 of the K-ras gene in DNA extracted from lung tumours and the matched sputum samples obtained from 22 lung cancer patients. K-ras mutations were identified in the lung tumours of 12 patients (54.5%) and in the sputum samples of 10 patients (45.5%). Nine patients showed an identical mutation in both the tumour and the matched sputum samples. There was a significant association between the presence of a K-ras mutation in a lung tumour and the detection of an identical mutation in the matched sputum sample of the lung cancer patient (kappa = 0.64, 95% confidence interval 0.32-0.95, p <0.01). K ras mutations were detected in sputum samples from cancer patients with all lung tumour grades, and both in the presence and the absence of lymph node metastasis. Therefore, K-ras mutations may provide useful diagnostic markers for lung cancer. PMID- 12775501 TI - DNA-protein crosslinks as a biomarker of exposure to solar radiation: a preliminary study in brick-kiln workers. AB - In India, fired clay bricks are produced in small-scale factories. There are 60, 000 active brick kilns, providing employment to nearly 12 million people in different suboccupations. This industry is largely non-mechanized and operates from November to June. Almost all the workers are exposed to direct sunlight for 8-10 h a day. Cellular DNA-protein crosslinks (DPCs) are the biologically active nucleoprotein complexes formed between DNA and proteins. Ultraviolet light and gamma-rays, and other suspected carcinogens in humans, induce DPC formation in blood cells. DPCs have therefore been identified as a biomarker for monitoring exposure to these hazardous agents. Here we report steady-state levels of DPCs in human peripheral lymphocytes from 46 brick-kiln workers exposed occupationally for 8-10 h a day to solar radiation in brickfields and 25 unexposed controls. A significant increase (p <0.05) in DPC content and DPC coefficients in peripheral lymphocytes was observed in the brick-kiln workers compared with the controls. The data suggest that the DPC content of lymphocytes could be a possible biomarker of exposure to solar radiation. However, further work is necessary to confirm this. PMID- 12775502 TI - Ultrapath XI: the academic mission--a view from the program organizer with a historical perspective. PMID- 12775503 TI - Bioterrorism and electron microscopic differentiation of poxviruses from herpesviruses: dos and don'ts. AB - With increased threat of terrorism, much attention is being directed toward readiness for biodefense. Smallpox virus, a deadly and much feared organism, is among possible bioterrorism agents. Herpesviruses, such as the one that causes chickenpox and shingles, produce skin lesions that may resemble those seen early in smallpox infection. Electron microscopy (EM) is a rapid and reliable method for differentiating poxviruses from herpesviruses. However, before becoming involved in the monitoring of potential smallpox cases, a laboratory must consider several issues, including expertise in virus identification, capacity for handling biohazards, and health and immune status of laboratory staff. PMID- 12775504 TI - Diagnostic pathology of microsporidiosis. AB - Microsporidia are ubiquitous spore-forming parasites that are important worldwide pathogens in the HIV/AIDS pandemic. They are also increasingly being seen in HIV( ) individuals. Infection has been documented in almost every tissue and organ in the body and in a broad spectrum of cell types, including epithelial, mesenchymal, and neural. Microsporidia elicit a wide range of pathology, e.g., inflammation and cell death, and symptoms, e.g., shortness of breath, sinusitis, and diarrhea with wasting. Untreated, microsporidiosis has been documented as a cause of death. PMID- 12775505 TI - Intranuclear rod myopathy, a rare and morphologically striking variant of nemaline rod myopathy. AB - A 4-year-old boy with muscle weakness underwent skeletal muscle biopsies. Light microscopy showed distinct eosinophilic inclusions within the majority of muscle cell nuclei, but none in the cytoplasm. Electron microscopy revealed crystalline, round to rod-shaped inclusions in the muscle cell nuclei. The inclusions stained positively for alpha-actinin. Intranuclear inclusions identical to those seen here have been described in rare cases of nemaline rod myopathy, though almost always together with classic intracytoplasmic rods. This case illustrates the importance of electron microscopy in the diagnosis of rare myopathies and in the characterization of cellular inclusions of unknown origin. PMID- 12775506 TI - Desmosomes and microvilli mean a lot: diagnosis of neoplasms of unknown origin using electron microscopy. AB - Neoplasms of unknown origin present a difficult diagnostic dilemma, particularly if they are very poorly differentiated. Adenocarcinomas, squamous cell carcinomas, melanomas, lymphomas, and sarcomas can all be very difficult to diagnose if the light microscopic cytomorphology is sufficiently undifferentiated. Electron microscopy (EM) can either demonstrate differentiation or narrow the range of differential diagnoses. The authors report the case of a 64-year-old male who has been HIV positive for several years and was found to have expansile lytic lesions in several ribs and a thumb fracture associated with a soft tissue mass which was biopsied. The tumor was composed of very pleomorphic malignant cells without specific differentiation. The malignant cells stained positive for pancytokeratin (AE 1/3), EMA, CEA, CK20, and CK7. Rare cells had mucicarmine-positive intracytoplasmic droplets. They were negative for S-100, calretinin, CD45, MART-1, and vimentin. EM revealed intracytoplasmic lumina with long microvilli and many well-formed desmosomal junctions. The diagnosis was initially very broad. Immunohistochemistry narrowed the diagnosis to carcinoma, but EM alone was able to narrow the diagnosis to poorly differentiated adenocarcinoma. In a neoplasm of unknown origin, EM can either narrow the differential significantly or, in the case of limited material, provide information that otherwise may not be attainable. PMID- 12775507 TI - Ultrastructural identification of primary tumor site from bone metastases. AB - The value of electron microscopy is demonstrated by ultrastructural identification of primary tumor site from bone metastases in 2 patients. PMID- 12775508 TI - Microwave and digital imaging technology reduce turnaround times for diagnostic electron microscopy. AB - The contributions of microwave methods and digital imaging techniques, when taken together, can reduce routine specimen processing and evaluation for diagnostic electron microscopy to a time frame never thought possible. Significant improvements in both technologies over the last 5 years led the authors to evaluate their combined attributes as the most likely candidate to provide a realistic solution in the reduction of turnaround times for diagnostic electron microscopy. For diagnostic electron microscopy to compete favorably with immunohistochemistry and other ancillary diagnostic techniques, it must improve its turnaround time. To evaluate this hypothesis the microwave-assisted processing results of over 2,000 diagnostic cases were evaluated as was a digital image administration system used for the acquisition and dissemination of diagnostic results. The incorporation of both technologies resulted in turnaround times being reduced to 4 h or less. PMID- 12775509 TI - Postirradiation epithelioid angiosarcoma of the breast: a case report with immunohistochemical and electron microscopic study. AB - A case of postirradiation epithelioid angiosarcoma of the breast in a 72-year-old woman is reported. She had had right breast conserving surgery, axillary lymph node dissection, and 50 Gy external beam radiation therapy for infiltrating ductal carcinoma. A skin lesion on the irradiated breast appeared 5 years after completion of radiation. Angiosarcoma was diagnosed in a contralateral axillary mass 8 months later. Light microscopically, the tumor was characterized by a sheet-like growth of epithelioid cells with focal vasoformative areas. Tumor cells were reactive for factor VIII-related antigen, cytokeratin and CD34. Electron microscopically, the tumor cells were round with smooth cell borders. They were closely apposed, occasionally forming a small lumen containing single red blood cells or aggregates of platelets. Groups of tumor cells were enclosed by an external lamina. The tumor cells had abundant cytoplasm with sparse organelles. Rare suggestive Weibel-Palade bodies were present. The immunohistochemical and ultrastructural findings in this postirradiation tumor were in agreement with previously reported findings in non-irradiation-induced epithelioid angiosarcomas. PMID- 12775510 TI - Bednar tumor: report of a case with immunohistochemical and ultrastructural study. AB - A slowly growing tumor in the right shoulder of a 38-year-old white male, which felt like a superficial cystic mass, was studied. The spindle cells, which represented the main component of the tumor, were arranged in a typical storiform pattern and were positive for CD34 and focally for CD117. The pigmented cells were mostly found at the center of the storiform whorls and were negative for S 100 protein and HMB-45. Ultrastructurally, the tumor consisted predominantly of nondescript mesenchymal spindle cells that resembled fibroblasts. The tumor cells blended into a loosely arranged stromal tissue background. The general appearance of pigmented cells was very similar to the nonpigmented spindle cells. The pigment appeared to be a mature form of melanin granules. The lack of premelanosomes, cell injections, basal lamina, and pinocytotic vesicles was inconsistent with a neural origin/neural differentiation hypothesis for this tumor. PMID- 12775511 TI - Male reproductive hormones and thyroid function in pesticide applicators in the Red River Valley of Minnesota. AB - In the present effort, 144 pesticide applicators and 49 urban control subjects who reported no chronic disease were studied. Applicators provided records of the season's pesticides used by product, volumes, dates, and methods of application. Blood specimens for examination of hormone levels were obtained in summer and fall. In the herbicide-only applicator group, significant increases in testosterone levels in fall compared to summer and also elevated levels of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) in the fall were noted. With respect to fungicide use, in an earlier cross-sectional epidemiologic study, data demonstrated that historic fungicide use was associated with a significant alteration of the sex ratio of children borne to applicators. As before, among current study subjects it was noted that historic fungicide use was associated with increased numbers of girls being born. Lower mean total testosterone concentrations by quartile were also correlated with increased numbers of live-born female infants. A downward summer to fall seasonal shift in thyroid-stimulating hormone (TSH) concentrations occurred among applicators but not among controls. Farmers who had aerial application of fungicides to their land in the current season showed a significant shift in TSH values (from 1.75 to 1.11 mU/L). Subclinical hypothyroidism was noted in 5/144 applicators (TSH values >4.5 mU/L), but not in urban control subjects. Based on current and past studies, it was concluded that, in addition to pesticide exposure, individual susceptibility and perhaps economic factors may play a supporting role in the reported results. PMID- 12775512 TI - Increased incidence of preterm delivery in mothers residing in an industrialized area in Taiwan. AB - The objective of this study was to determine whether women living in the vicinity of industrial districts where emission levels of air contaminants from multiple sources including petrochemical, petroleum, steel, and shipbuilding industries are highly correlated with increased incidence of preterm births. The prevalence of delivery of preterm-birth infants was significantly higher in mothers living in the industrial study area compared to control regions of Taiwan. After controlling for several possible confounders (including maternal age, season, marital status, maternal education, infant gender, and birthplace), the adjusted odds ratio was 1.11 (95% CI = 1.02-1.21) for delivery of preterm infants in the industrial area. Data provide further support for the hypothesis that air pollution can affect the outcome of pregnancy, although a semiecological study cannot confirm a direct causal relationship. PMID- 12775514 TI - Asphalt exposure enhances neuropeptide levels in sensory neurons projecting to the rat nasal epithelium. AB - Asphalt fumes have been reported to produce nasal irritation in road workers. Since inhaled irritants can increase substance P (SP) production in airway neurons, the effects of asphalt fumes on SP production in trigeminal ganglia (TG) sensory neurons innervating the nasal mucosa were investigated. The effects of asphalt fumes on nasal mucosal innervation were examined by measuring SP and calcitonin-gene-related peptide (CGRP) levels in rat TG neurons projecting to the nasal epithelium. Female Sprague-Dawley rats were exposed to asphalt fumes at 16.0 +/- 8.1mg /m3 for 5 consecutive days, 3.5 h/d. Inflammatory cells were measured in nasal cavity lavage fluid. SP and CGRP immunoreactivity (IR) was measured in the cell bodies of trigeminal ganglion sensory neurons projecting to the nasal cavity. A significant increase in neutrophils and macrophages was observed after asphalt fume exposure indicating an inflammatory response in the nasal cavity. The percentage of SP-IR neurons increased significantly in the asphalt-exposed rats, and the proportion of CGRP-IR neurons was also elevated following asphalt exposure. These results indicate that exposure to asphalt fumes produces inflammation and increases the levels of SP and CGRP in TG neurons projecting to the nasal epithelium. The findings are consistent with asphalt induced activation of sensory C-fibers in the nasal cavity. Enhanced sensory neuropeptide release from nerve terminals in the nasal cavity may produce neurogenic inflammation associated with nasal irritation following exposure to asphalt fumes. PMID- 12775513 TI - Response of alveolar macrophages from inducible nitric oxide synthase knockout or wild-type mice to an in vitro lipopolysaccharide or silica exposure. AB - The role of nitric oxide (NO) in pulmonary disease has been controversial with both antiinflammatory (scavenging radicals and inhibiting NF-eB activation) and proinflammatory (forming highly reactive peroxynitrite and augmenting NF-eB activation by inflammatory agents) actions reported. Therefore, a study has been initiated to determine whether deletion of the inducible nitric oxide synthase (iNOS) gene in the C57BL/6J mouse alters the pulmonary macrophage response to lipopolysaccharide (LPS) or silica. The objective of the initial phase of this study was to determine the difference in responsiveness of alveolar macrophages (AMs), harvested from naive wild-type (WT) or iNOS knockout (iNOS KO) mice, to an in vitro LPS or silica exposure. Primary AMs were obtained by bronchoalveolar lavage (BAL) from age- and weight-matched iNOS KO and WT mice. The cells were treated with interferon-gamma (IFN-a) (50 U/ml), IFN-a (50 U/ml) + LPS (1 microg/ml), LPS (0.01-100 microg/ml), or silica (25-250 microg/ml). The following parameters were measured: nitrate and nitrite (NOx), tumor necrosis factor-a (TNF a), macrophage inflammatory protein-2 (MIP-2), intracellular generation of the reactive oxygen species (ROS) hydrogen peroxide (H(2)O(2) and superoxide (O(* 2)), and basal (unstimulated) total antioxidant capacity. Data show a significant increase in NOx production upon exposure to IFN-a +/- LPS in the WT but not iNOS KO AMs. NOx production by iNOS KO or WT AMs was not altered by in vitro exposure to LPS or silica alone. LPS, but not silica, induced TNF-a and MIP-2 production in both iNOS KO and WT AMs. Statistical analysis of concentration response curves found a significant tendency for greater mediator production in the iNOS KO versus WT AMs. Basal intracellular production of H(2)O(2) and O(*- 2) was significantly greater in the iNOS KO compared to WT AMs. In contrast, LPS- (10 microg/ml) or silica- (100 microg/ml) stimulated intracellular oxidant production was lower in iNOS KO AMs, but overall (basal + stimulated) inflammatory capacity was similar between the cell types. The basal total antioxidant production of the iNOS KO AMs was approximately twofold higher than the WT AMs. In conclusion, certain compensatory changes appear to occur in AMs from iNOS KO mice. In response to the inability to induce NO production, iNOS KO AMs exhibit significantly higher basal generation of H(2)O(2) and (O(*- 2)) as well as higher total antioxidant levels. In addition, LPS induced TNF-a and MIP-2 production tend to be higher in AMs from iNOS KO mice. Such compensatory changes in the AM response may affect the response of iNOS KO mice to inflammatory exposures. PMID- 12775515 TI - The uptake, distribution, metabolism, and excretion of methyl tertiary-butyl ether inhaled alone and in combination with gasoline vapor. AB - The purpose of these studies was to evaluate the tissue uptake, distribution, metabolism, and excretion of methyl tertiary-butyl ether (MTBE) in rats and to determine the effects of coinhalation of the volatile fraction of unleaded gasoline on these parameters. Male F344 rats were exposed nose-only once for 4 h to 4, 40, or 400 ppm 14C-MTBE and to 20 and 200 ppm of the light fraction of unleaded gasoline (LFG) containing 4 and 40 ppm 14C-MTBE, respectively. To evaluate the effects of repeated inhalation of LFG on the fate of inhaled MTBE, rats were exposed for 7 consecutive days to 20 and 200 ppm LFG followed on d 8 by exposure to LFG containing 14C-MTBE. Three subgroups of rats were included for evaluation of respiratory parameters, rates and routes of excretion, and tissue distribution and elimination. MTBE and its chief metabolite, tertiary-butyl alcohol, were quantitated in blood and kidney (immediately after exposure), and the major urinary metabolites, 2-hydroxyisobutyric acid and 2-methyl-1,2- propanediol, were identified and quantified in urine. Inhalation of MTBE alone or as a component of LFG had no concentration-dependent effect on respiratory minute volume. The initial body burdens (IBBs) of MTBE equivalents achieved after 4 h of exposure to MTBE did not increase linearly with exposure concentration. MTBE equivalents rapidly distributed to all tissues examined, with the largest percentages distributed to liver. Between 40 and 400 ppm, there was a significant reduction in percentage of the IBB present in the major organs examined, both immediately and 72 h after exposure. At 400 ppm, the elimination rates of MTBE equivalents from tissues changed significantly. Furthermore, at 400 ppm there was a significant decrease in the elimination half-time of volatile organic compounds (VOCs) in breath and a significant increase in the percentage of the IBB of MTBE equivalents eliminated as VOCs in breath. LFG coexposure significantly decreased the percentage of the MTBE equivalent IBBs in tissues and increased rates of elimination of MTBE equivalents. The study results indicate that the uptake and fate of inhaled MTBE are altered upon increasing exposure levels from 4 to 400 ppm, suggesting that toxic effects observed previously upon repeated inhalation of concentrations of 400 ppm or greater may not necessarily be linearly extrapolated to effects that might occur at lower concentrations. Furthermore, coexposure to LFG, whether acute or repeated, decreases tissue burdens of MTBE equivalents and enhances the elimination rate of MTBE and its metabolites, thereby potentially reducing the toxic effects of the MTBE compared to when it is inhaled alone. PMID- 12775516 TI - Health of tree swallow (Tachycineta bicolor) nestlings exposed to chlorinated hydrocarbons in the St Lawrence River basin. Part I. Renal and hepatic vitamin A concentrations. AB - Sixteen-day-old tree swallows (Tachycineta bicolor), near fledging, were collected in 1999 and 2000 from nine sites representing a gradient of dioxin concentrations, within the vicinity of the St. Lawrence River in Canada and the United States, to determine if organochlorine contaminants correlated with vitamin A levels measured as retinol and retinyl palmitate. Mean concentrations of hepatic retinol ranged from 3 mg /kg to 13 mg /kg, and from 0.35 mg /kg to 1.5 mg /kg for renal retinol. Mean concentrations of hepatic retinyl palmitate ranged from 18 mg /kg to 146 mg /kg, and 1mg /kg to 6 mg/kg for renal retinyl palmitate. In 1999, molar ratio of renal retinol: retinyl palmitate was significantly and positively correlated with total polychlorinated dibenzodioxin (PCDD) concentration. Among sites, total PCDDs ranged from 5.4 ng /kg wet weight to 79.5 ng /kg wet weight in tree swallows. These results suggest that current levels of organochlorine contaminants in the St. Lawrence River and surrounding tributaries may be interacting with the vitamin A pathway. Lower circulating levels and higher tissue concentrations of retinoids may result in compromised immune function and reduced reproductive success in adult birds. PMID- 12775518 TI - A review of mercury in Lake Victoria, East Africa: implications for human and ecosystem health. AB - Lake Victoria, East Africa, has been the site of many recent studies measuring mercury (Hg) concentrations in water, fish, sediment, soil, and humans. Most of these studies were motivated by concerns about Hg contamination from processing of gold ore on the southern shores. Total Hg (THg) concentrations in fish were usually below permissible World Health Organization (WHO) concentrations and international marketing limits and do not threaten the lucrative export industry. Nile perch 3-10 kg and most >10 kg had THg concentrations above the WHO threshold concentrations for at-risk groups (200 ng/g). Elevated THg concentrations in large Nile perch are not of major concern because Nile perch are rarely consumed by the people living on Lake Victoria and very large Nile perch are becoming increasingly rare in catches. Water THg concentrations were below Canadian drinking water guidelines but were elevated relative to those in the northern Great Lakes. Sediment and soil THg concentrations were within inter-national guidelines and are comparable to those in northern latitudes but are lower than those in the Amazon basin. Biomass burning and soil erosion are estimated to be the major sources of THg for the lake and probably constitute a larger source of THg than gold mining in Tanzania.THg concentrations in urine and hair from human volunteers indicate that while gold miners and frequent skin-bleaching cream users are at risk of inorganic mercury poisoning, the rest of the population, including fishermen, is not. Human exposure assessments demonstrated that fish consumption and soil geophagy constitute major sources of THg for humans, but the total estimated daily intake of THg was below the Health Canada tolerable daily intake (TDI) limits. The use of beauty creams containing high inorganic Hg concentrations, however, caused the estimated THg exposure to exceed the TDI. The high THg content in the hair of regular cream users supports this assessment. The nutritional benefits of fish and soil geophagy outweigh the risk of THg poisoning. Still, due to the importance of those natural items as a THg source to humans, as well as the changing nature of Lake Victoria, regular monitoring and risk assessments need to be carried out in the Lake Victoria catchment. PMID- 12775517 TI - Reproductive system impairment of mice fed diets containing beluga whale blubber from the St Lawrence estuary and arctic populations. AB - The toxic potential of naturally relevant mixtures of PCBs and other organohalogens on the reproductive system of C57Bl/6 female mice was assessed. Mice were fed diets in which lipids were replaced by blubber of beluga whales from a highly contaminated population of the Saint Lawrence River, and a less contaminated population from the Arctic Ocean. Ratios of blubber from both sources were mixed in order to perform a dose-response study. Control mice were fed diets for 90 d in which fat was replaced by corn oil or beef tallow. There were no significant effects of diets on body, liver, spleen or thymus weights. Similarly ovulation occurred in all control and experimental groups. However, Graafian follicles from ovaries of mice fed contaminated diets showed abnormal development of oocytes. Cumulus granulosa cells bind normally to the oocyte prior to ovulation and are essential for sperm penetration and fertilization. These cells were absent in both Graafian follicles and ovulated oocytes in the oviduct of all groups fed contaminated diets. Oviducts of these mice revealed evidence of epithelial degeneration. These results suggest the female mouse reproductive system is sensitive to organohalogens and illustrate the toxic potential of contaminant mixtures as found in the less contaminated Arctic population. PMID- 12775519 TI - Biological and health effects of exposure to kerosene-based jet fuels and performance additives. AB - Over 2 million military and civilian personnel per year (over 1 million in the United States) are occupationally exposed, respectively, to jet propulsion fuel-8 (JP-8), JP-8 +100 or JP-5, or to the civil aviation equivalents Jet A or Jet A-1. Approximately 60 billion gallon of these kerosene-based jet fuels are annually consumed worldwide (26 billion gallon in the United States), including over 5 billion gallon of JP-8 by the militaries of the United States and other NATO countries. JP-8, for example, represents the largest single chemical exposure in the U.S. military (2.53 billion gallon in 2000), while Jet A and A-1 are among the most common sources of nonmilitary occupational chemical exposure. Although more recent figures were not available, approximately 4.06 billion gallon of kerosene per se were consumed in the United States in 1990 (IARC, 1992). These exposures may occur repeatedly to raw fuel, vapor phase, aerosol phase, or fuel combustion exhaust by dermal absorption, pulmonary inhalation, or oral ingestion routes. Additionally, the public may be repeatedly exposed to lower levels of jet fuel vapor/aerosol or to fuel combustion products through atmospheric contamination, or to raw fuel constituents by contact with contaminated groundwater or soil. Kerosene-based hydrocarbon fuels are complex mixtures of up to 260+ aliphatic and aromatic hydrocarbon compounds (C(6) -C(17+); possibly 2000+ isomeric forms), including varying concentrations of potential toxicants such as benzene, n-hexane, toluene, xylenes, trimethylpentane, methoxyethanol, naphthalenes (including polycyclic aromatic hydrocarbons [PAHs], and certain other C(9)-C(12) fractions (i.e., n-propylbenzene, trimethylbenzene isomers). While hydrocarbon fuel exposures occur typically at concentrations below current permissible exposure limits (PELs) for the parent fuel or its constituent chemicals, it is unknown whether additive or synergistic interactions among hydrocarbon constituents, up to six performance additives, and other environmental exposure factors may result in unpredicted toxicity. While there is little epidemiological evidence for fuel-induced death, cancer, or other serious organic disease in fuel-exposed workers, large numbers of self-reported health complaints in this cohort appear to justify study of more subtle health consequences. A number of recently published studies reported acute or persisting biological or health effects from acute, subchronic, or chronic exposure of humans or animals to kerosene-based hydrocarbon fuels, to constituent chemicals of these fuels, or to fuel combustion products. This review provides an in-depth summary of human, animal, and in vitro studies of biological or health effects from exposure to JP-8, JP-8 +100, JP-5, Jet A, Jet A-1, or kerosene. PMID- 12775520 TI - 18S ribosomal RNA and tetrapod phylogeny. AB - Previous phylogenetic analyses of tetrapod 18S ribosomal RNA (rRNA) sequences support the grouping of birds with mammals, whereas other molecular data, and morphological and paleontological data favor the grouping of birds with crocodiles. The 18S rRNA gene has consequently been considered odd, serving as "definitive evidence of different genes providing significantly different estimates of phylogeny in higher organisms" (p. 156; Huelsenbeck et al., 1996, Trends Ecol. Evol. 11:152-158). Our research indicates that the previous discrepancy of phylogenetic results between the 18S rRNA gene and other genes is caused mainly by (1) the misalignment of the sequences, (2) the inappropriate use of the frequency parameters, and (3) poor sequence quality. When the sequences are aligned with the aide of the secondary structure of the 18S rRNA molecule and when the frequency parameters are estimated either from all sites or from the variable domains where substitutions have occurred, the 18S rRNA sequences no longer support the grouping of the avian species with the mammalian species. PMID- 12775521 TI - The value of idiosyncratic markers and changes to conserved tRNA sequences from the mitochondrial genome of hard ticks (Acari: Ixodida: Ixodidae) for phylogenetic inference. AB - Idiosyncratic markers are features of genes and genomes that are so unusual that it is unlikely that they evolved more than once in a lineage of organisms. Here we explore further the potential of idiosyncratic markers and changes to typically conserved tRNA sequences for phylogenetic inference. Hard ticks were chosen as the model group because their phylogeny has been studied extensively. Fifty-eight candidate markers from hard ticks (family Ixodidae) and 22 markers from the subfamily Rhipicephalinae sensu lato were mapped onto phylogenies of these groups. Two of the most interesting markers, features of the secondary structure of two different tRNAs, gave strong support to the hypothesis that species of the Prostriata (Ixodes spp.) are monophyletic. Previous analyses of genes and morphology did not strongly support this relationship, instead suggesting that the Prostriata is paraphyletic with respect to the Metastriata (the rest of the hard ticks). Parallel or convergent evolution was not found in the arrangements of mitochondrial genes in ticks nor were there any reversals to the ancestral arthropod character state. Many of the markers identified were phylogenetically informative, whereas others should be informative with study of additional taxa. Idiosyncratic markers and changes to typically conserved nucleotides in tRNAs that are phylogenetically informative were common in this data set, and thus these types of markers might be found in other organisms. PMID- 12775522 TI - Phylogeny and evolution of calcareous sponges: monophyly of calcinea and calcaronea, high level of morphological homoplasy, and the primitive nature of axial symmetry. AB - Because calcareous sponges are triggering renewed interest with respect to basal metazoan evolution, a phylogenetic framework of their internal relationships is needed to clarify the evolutionary history of key morphological characters. Morphological variation was scored at the suprageneric level within Calcispongia, but little phylogenetic information could be retrieved from morphological characters. For the main subdivision of Calcispongia, the analysis of morphological data weakly supports a classification based upon cytological and embryological characters (Calcinea/Calcaronea) rather than the older classification scheme based upon the aquiferous system (Homocoela/Heterocoela). The 18S ribosomal RNA data were then analyzed, both alone and in combination with morphological characters. The monophyly of Calcispongia is highly supported, but the position of this group with respect to other sponge lineages and to eumetazoan taxa is not resolved. The monophyly of both Calcinea and Calcaronea is retrieved, and the data strongly rejected the competing Homocoela/Heterocoela hypothesis. The phylogeny implies that characters of the skeleton architecture are highly homoplastic, as are characters of the aquiferous system. However, axial symmetry seems to be primitive for all Calcispongia, a conclusion that has potentially far-reaching implications for hypotheses of early body plan evolution in Metazoa. PMID- 12775523 TI - The systematic utility of floral and vegetative fragrance in two genera of nyctaginaceae. AB - We examined relationships between fragrance and phylogeny using a number of approaches to coding fragrance data and comparing the hierarchical information in fragrance data with the phylogenetic signal in a DNA sequence data set. We first used distance analyses to determine which coding method(s) best distinguishes species while grouping conspecifics. Results suggest that interspecific differences in fragrance composition were maximized by coding as presence/absence of fragrance compounds and biosynthetic pathways rather than when quantitative information was also included. Useful systematic information came from both compounds and pathways and from fragrance emitted by both floral and vegetative tissues. The coding methods that emerged from the distance analyses as best distinguishing species were then adapted for use in phylogenetic analysis. Although hierarchical signal among fragrance data sets was congruent, this signal was highly incongruent with the phylogenetic signal in the DNA sequence data. Notably, topologies inferred from fragrance data sets were congruent with the DNA topology only in the most distal portions (e.g., sister group pairs or closely related species that had similar fragrance profiles were often recovered by analyses of fragrance). Examination of consistency and retention indices for individual fragrance compounds and pathways as optimized onto one of the most parsimonious trees inferred from DNA data revealed that although most compounds were homoplastic, some compounds were perfectly congruent with the DNA phylogeny. In particular, compounds and pathways found in a few taxa were less homoplastic than those found in many taxa. Pathways that synthesize few volatiles also seem to have lower homoplasy than those that produce many. Although fragrance data as a whole may not be useful in phylogeny reconstruction, these data can provide additional support for clades reconstructed with other types of characters. Factors other than phylogeny, including pollinator interactions, also likely influence fragrance composition. PMID- 12775524 TI - Failed refutations: further comments on parsimony and likelihood methods and their relationship to Popper's degree of corroboration. AB - Kluge's (2001, Syst. Biol. 50:322-330) continued arguments that phylogenetic methods based on the statistical principle of likelihood are incompatible with the philosophy of science described by Karl Popper are based on false premises related to Kluge's misrepresentations of Popper's philosophy. Contrary to Kluge's conjectures, likelihood methods are not inherently verificationist; they do not treat every instance of a hypothesis as confirmation of that hypothesis. The historical nature of phylogeny does not preclude phylogenetic hypotheses from being evaluated using the probability of evidence. The low absolute probabilities of hypotheses are irrelevant to the correct interpretation of Popper's concept termed degree of corroboration, which is defined entirely in terms of relative probabilities. Popper did not advocate minimizing background knowledge; in any case, the background knowledge of both parsimony and likelihood methods consists of the general assumption of descent with modification and additional assumptions that are deterministic, concerning which tree is considered most highly corroborated. Although parsimony methods do not assume (in the sense of entailing) that homoplasy is rare, they do assume (in the sense of requiring to obtain a correct phylogenetic inference) certain things about patterns of homoplasy. Both parsimony and likelihood methods assume (in the sense of implying by the manner in which they operate) various things about evolutionary processes, although violation of those assumptions does not always cause the methods to yield incorrect phylogenetic inferences. Test severity is increased by sampling additional relevant characters rather than by character reanalysis, although either interpretation is compatible with the use of phylogenetic likelihood methods. Neither parsimony nor likelihood methods assess test severity (critical evidence) when used to identify a most highly corroborated tree(s) based on a single method or model and a single body of data; however, both classes of methods can be used to perform severe tests. The assumption of descent with modification is insufficient background knowledge to justify cladistic parsimony as a method for assessing degree of corroboration. Invoking equivalency between parsimony methods and likelihood models that assume no common mechanism emphasizes the necessity of additional assumptions, at least some of which are probabilistic in nature. Incongruent characters do not qualify as falsifiers of phylogenetic hypotheses except under extremely unrealistic evolutionary models; therefore, justifications of parsimony methods as falsificationist based on the idea that they minimize the ad hoc dismissal of falsifiers are questionable. Probabilistic concepts such as degree of corroboration and likelihood provide a more appropriate framework for understanding how phylogenetics conforms with Popper's philosophy of science. Likelihood ratio tests do not assume what is at issue but instead are methods for testing hypotheses according to an accepted standard of statistical significance and for incorporating considerations about test severity. These tests are fundamentally similar to Popper's degree of corroboration in being based on the relationship between the probability of the evidence e in the presence versus absence of the hypothesis h, i.e., between p(e|hb) and p(e|b), where b is the background knowledge. Both parsimony and likelihood methods are inductive in that their inferences (particular trees) contain more information than (and therefore do not follow necessarily from) the observations upon which they are based; however, both are deductive in that their conclusions (tree lengths and likelihoods) follow necessarily from their premises (particular trees, observed character state distributions, and evolutionary models). For these and other reasons, phylogenetic likelihood methods are highly compatible with Karl Popper's philosophy of science and offer several advantages over parsimony methods in this context. PMID- 12775525 TI - Accelerated likelihood surface exploration: the likelihood ratchet. AB - The existence of multiple likelihood maxima necessitates algorithms that explore a large part of the tree space. However, because of computational constraints, stepwise addition-based tree-searching methods do not allow for this exploration in reasonable time. Here, I present an algorithm that increases the speed at which the likelihood landscape can be explored. The iterative algorithm combines the computational speed of distance-based tree construction methods to arrive at approximations of the global optimum with the accuracy of optimality criterion based branch-swapping methods to improve on the result of the starting tree. The algorithm moves between local optima by iteratively perturbing the tree landscape through a process of reweighting randomly drawn samples of the underlying sequence data set. Tests on simulated and real data sets demonstrated that the optimal solution obtained using stepwise addition-based heuristic searches was found faster using the algorithm presented here. Tests on a previously published data set that established the presence of tree islands under maximum likelihood demonstrated that the algorithm identifies the same tree islands in a shorter amount of time than that needed using stepwise addition. The algorithm can be readily applied using standard software for phylogenetic inference. PMID- 12775526 TI - Ancient allopolyploid speciation in Geinae (Rosaceae): evidence from nuclear granule-bound starch synthase (GBSSI) gene sequences. AB - A nuclear low-copy gene phylogeny provides strong evidence for the hybrid origin of seven polyploid species in Geinae (Rosaceae). In a gene tree, alleles at homologous loci in an allopolyploid species are expected to be sisters to orthologues in the ancestral taxa rather than to each other. Alleles at a duplicated locus in an autopolyploid, however, are expected to be more closely related to each other than they are to any orthologous copies in closely related species. We cloned and sequenced about 1.9 kilobases from the 5' end of the GBSSI 1 gene from two diploid, one tetraploid, and six hexaploid species. Each of the three loci in the hexaploid species forms a separate group, two of which are more closely related to copies in other species than they are to each other. This finding indicates that the hexaploid lineage evolved through two consecutive allopolyploidization events. Based on the GBSSI-1 gene tree, we hypothesized that there was an initial hybridization between a diploid species from the ancestral lineage of Coluria and Waldsteinia and an unknown diploid species to form the tetraploid Geum heterocarpum lineage. Backcrossing of G. heterocarpum with a representative of the unknown diploid lineage then resulted in a hexaploid lineage that has radiated considerably since its origin, comprising at least 40 extant species with various morphologies. A penalized likelihood analysis indicated that Geinae may be about 17 million years old, implying that the hypothesized allopolyploid speciation events are relatively ancient. Six of the 22 cloned Geinae GBSSI-1 copies in this study, which all are duplicate copies in polyploid taxa, may have become pseudogenes. We compared the GBSSI-1 phylogeny with one from chloroplast data and explored implications for the evolution of some fruit characters. PMID- 12775527 TI - True and false gharials: a nuclear gene phylogeny of crocodylia. AB - The phylogeny of Crocodylia offers an unusual twist on the usual molecules versus morphology story. The true gharial (Gavialis gangeticus) and the false gharial (Tomistoma schlegelii), as their common names imply, have appeared in all cladistic morphological analyses as distantly related species, convergent upon a similar morphology. In contrast, all previous molecular studies have shown them to be sister taxa. We present the first phylogenetic study of Crocodylia using a nuclear gene. We cloned and sequenced the c-myc proto-oncogene from Alligator mississippiensis to facilitate primer design and then sequenced an 1,100-base pair fragment that includes both coding and noncoding regions and informative indels for one species in each extant crocodylian genus and six avian outgroups. Phylogenetic analyses using parsimony, maximum likelihood, and Bayesian inference all strongly agreed on the same tree, which is identical to the tree found in previous molecular analyses: Gavialis and Tomistoma are sister taxa and together are the sister group of Crocodylidae. Kishino-Hasegawa tests rejected the morphological tree in favor of the molecular tree. We excluded long-branch attraction and variation in base composition among taxa as explanations for this topology. To explore the causes of discrepancy between molecular and morphological estimates of crocodylian phylogeny, we examined puzzling features of the morphological data using a priori partitions of the data based on anatomical regions and investigated the effects of different coding schemes for two obvious morphological similarities of the two gharials. PMID- 12775529 TI - Evaluation of the strategy of long-branch subdivision to improve the accuracy of phylogenetic methods. PMID- 12775528 TI - Combined support for wholesale taxic atavism in gavialine crocodylians. AB - Morphological and molecular data sets favor robustly supported, contradictory interpretations of crocodylian phylogeny. A longstanding perception in the field of systematics is that such significantly conflicting data sets should be analyzed separately. Here we utilize a combined approach, simultaneous analyses of all relevant character data, to summarize common support and to reconcile discrepancies among data sets. By conjoining rather than separating incongruent classes of data, secondary phylogenetic signals emerge from both molecular and morphological character sets and provide solid evidence for a unified hypothesis of crocodylian phylogeny. Simultaneous analyses of four gene sequences and paleontological data suggest that putative adaptive convergences in the jaws of gavialines (gavials) and tomistomines (false gavials) offer character support for a grouping of these taxa, making Gavialinae an atavistic taxon. Simple new methods for measuring the influence of extinct taxa on topological support indicate that in this vertebrate order fossils generally stabilize relationships and accentuate hidden phylogenetic signals. Remaining inconsistencies in minimum length trees, including concentrated hierarchical patterns of homoplasy and extensive gaps in the fossil record, indicate where future work in crocodylian systematics should be directed. PMID- 12775530 TI - The taxonomic impediment overcome: NSF's Partnerships for Enhancing Expertise in Taxonomy (PEET) as a model. PMID- 12775532 TI - Solid and papillary epithelial neoplasm of the pancreas in an 11-year-old girl: case report and literature review. AB - An 11-year-old girl presented with episodic abdominal pain of 2 years' duration. CT scan of the abdomen showed a mass in the tail of the pancreas. A distal pancreatectomy was done and the tumor was excised. Macroscopic and immunohistochemical studies were compatible with a solid and papillary epithelial neoplasm. This is a rare neoplasm with a decidedly female predominance. It has a very low malignant potential with a good prognosis. Surgical removal of the tumor is usually curative. PMID- 12775533 TI - Treatment of essential thrombocythemia in childhood. AB - Essential thrombocythemia is a rare myeloproliferative disorder in childhood. For symptomatic patients with platelet counts greater than 1000 x 10(9)/L, cytoreductive treatment is recommended. The authors describe a 5-year-old boy with symptomatic essential thrombocythemia who was treated with anagrelide (Agrylin) for 23 months. He responded well but early on developed anemia. Because anagrelide is thought to specifically inhibit thrombopoesis, anemia is assumed to be a rather rare and late adverse effect. Its early occurence in this patient might indicate an increased vulnerability to anagrelide (Agrylin) in young children. PMID- 12775534 TI - Primary hemophagocytic lymphohistiocytosis in Turkish children. AB - Nineteen children with hemophagocytic lymphohistiocytosis (HLH) were studied in the Department of Pediatric Hematology, Hacettepe University. Patients were divided into two groups. Group 1: Thirteen patients were classified as having a genetic etiology (7 familial, 6 presumed familial) on the basis of an affected sibling and consanguinity. There was a history of consanguineous marriage in 13 of the families. Seven of them had a history of a sibling with HLH. Group 2: Six patients were diagnosed with sporadic HLH. The age at presentation for familial patients was 0.7-84 months (mean 21.9 +/- 24.9 months), and for sporadic cases it was 2.5-48 months (mean 22.7 +/- 19.8 months). The clinical and laboratory data of these two groups were similar at diagnosis. Thirteen cases were diagnosed premortem by bone marrow aspiration. Splenic biopsy was performed in 2 patients. Four patients were diagnosed by postmortem examination. Elevated LDH levels were found in all patients tested. No significant differences for clinical and laboratory data were found between the two groups. PMID- 12775536 TI - The combined analysis of P-glycoprotein expression and activity predicts outcome in childhood acute lymphoblastic leukemia. AB - The link between drug resistance and relapse was often suggested, but rarely demonstrated in long-range clinical studies. Since it is nowadays recommended to validate immunocytochemical results, the authors studied prospectively 52 acute lymphoblastic leukemia (ALL) patients with an immunocytochemical test and a functional flow cytometric test. The 4-year EFS and OS were 79.3% and 85.2%, respectively. Patients scoring positive in both tests had a significantly higher relapse rate and worse survival (log rank p = 0.007 and 0.047 for event-free survival and overall survival, respectively). Among the different prognostic variables evaluated, only the combination of P-gp expression and activity was a statistically significant parameter predicting relapse in childhood ALL. PMID- 12775535 TI - The role of short course of high-dose methylprednisolone in children with acute myeloblastic leukemia (FAB M2) presented with myeloid tumor. AB - The authors have previously demonstrated a favorable effect of high-dose methylprednisolone (HDMP), which can induce differentiation and apoptosis of leukemic cells in children with acute myeloblastic leukemia (AML). Here, they evaluate the effect of short-course HDMP in 2 children with acute myeloblastic leukemia (AML-M2) presented with myeloid tumor (MT). Methylprednisolone (20 or 30 mg/kg/day) was given orally, in a single dose, without using other antileukemic agents. Rapid cytoreduction in MT, peripheral blood, and bone marrow blasts was observed in both children following short-course (4 or 7 days) HDMP treatment, possibly due to HDMP-induced differentiation and apoptosis of leukemic cells. The effects of HDMP should be explored in patients with other subtypes of AML who present with MT. PMID- 12775537 TI - Neonatal alloimmune thrombocytopenia may be less severe in a subsequent pregnancy. AB - Neonatal alloimmune thrombocytopenia (NAIT) is more common than previously appreciated. Clinical presentation is varied, but it is generally a severe disorder and it is said to be more severe in subsequent pregnancies. The authors present a case of fatal disseminated intravascular coagulation that was diagnosed as NAIT only after presentation of a subsequent surviving sibling with intracranial hemorrhage and thrombocytopenia. Despite the perinatal complications, this second sibling had good neurodevelopmental outcome. This is the first reported case of less severe disease occurring in a subsequent pregnancy. PMID- 12775538 TI - Primary high-grade non-Hodgkin lymphoma of the trachea in an adolescent. AB - Lymphomas belong to the most treatable and curable malignant tumors of childhood. They should always be considered among differential diagnosis, even in unusual locations, and tumor material should be processed adequately to reach true diagnosis. The authors report on a 16-year-old adolescent with a tracheal lymphoma, a rare location of lymphoma in childhood. Because only formalin-fixed tumor material was obtained, pathology showed two differential diagnoses and the entity could never be fully clarified as anaplastic plasmacytoma or plasmoblastic lymphoma. Thus, full lymphoma diagnostic workup should be performed even in tumors with atypical location. PMID- 12775539 TI - Spontaneous fatal intracranial hemorrhage in a child with Evans syndrome. AB - Evans syndrome is a chronic and recurrent disease often resistant to multiagent therapy. Spontaneous intracranial hemorrhage is a rare but life-threatening complication in children with Evans syndrome. The authors report the fatal course of a child with chronic thrombocytopenia who presented a spontaneous intracranial hemorrhage. The case indicates that this potentially fatal event may occur even in children with a moderate grade of chronic thrombocytopenia. Early recognition and aggressive management is required for an adequate management of this rare complication. PMID- 12775540 TI - Early manifestation of Ghosal-type hemato-diaphyseal dysplasia. AB - Ghosal-type hemato-diaphyseal dysplasia is a rare autosomal recessive disorder with distinctive diaphyseal and metaphyseal dysplasia of long bones and steroid dependant anemia. The authors describe a 20-month-old girl who had had a severe transfusion-dependent anemia since late infancy and marked locomotion difficulties as a toddler. The diagnosis was established by X-ray bone survey. The anemia was treated with oral prednisolone. Since then, the patient has been doing well on steroid-maintenance therapy and has no more walking difficulties. The incidence of hemato-diaphyseal dysplasia in the Indian subcontinent and Middle East is notable. PMID- 12775541 TI - Laparoscopic adnexectomy of a persistent ovarian tumor in a girl with acute lymphoblastic leukemia relapse. AB - The management of children with acute lymphoblastic leukemia (ALL) relapse and an ovarian tumor remains controversial. The authors report about a 4-year-old girl who developed a late bone marrow and cutaneous relapse of her pre-B-cell ALL and revealed an enlargement of her left ovary (4 x 3 x 2 cm). Chemotherapy (ALL-REZ BFM pilot-protocol 2002) achieved effective remission, but the ovarian mass depicted no regression. Laparoscopic adnexectomy was performed and the tumor could be extracted in a specimen-bag through a 12-mm umbilical incision. Histology detected no viable lymphoblasts, but a fibrotic enlargement due to previous cellular infiltration. The authors conclude that in children with ALL relapse and an ovarian tumor, malignant infiltration as well as local response to chemotherapy can be judged only by surgical excision and histopathologic examination. Laparoscopic oophoradnexectomy is a valuable management option in these children. PMID- 12775542 TI - Acute vulvar ulceration as the main presenting manifestation of hemophagocytic syndrome. AB - A 14-year-old female exhibited an acute vulvar ulcer during the course of hemophagocytic syndrome (HPS). The patient presented persistent high fever and a deep painful vulvar ulcer lasting for more than 2 weeks. Neither infection with sexually transmitted agents nor autoimmune disorder were found to be positive. The presence of hemophagocytosis in the bone marrow and elevation of urinary beta 2-microglobulin (beta-2M) prompted the diagnosis of HPS. Acute vulvar ulcer is rare, but it should be recognized as a mucous manifestation of HPS. During the clinical course, urinary beta-2M was the most sensitive marker for the evaluation of the disease activity of HPS. PMID- 12775543 TI - Inadvertent intramuscular administration of vincristine: lack of untoward effects without any treatment except administration of hot compresses. AB - A seven-year-old boy with acute lymphoblastic leukemia received vincristine sulphate 1 mg, 1 ml intramuscularly, into his glutea, inadvertently, in the local hospital. The mother, applied hot compresses for 16 hours, starting 6.5 hours after the injection. Then, she told, the slight pain and the reddened area which developed around the injection site disappeared completely and he turned back to his daily activities. His physical examination, 2 weeks after the injection and during his follow-up revealed no abnormality. PMID- 12775545 TI - Mental health courts as an alternative to criminal courts for those who have mental illness and have been charged with a crime. PMID- 12775547 TI - Integrating aesthetics into advanced practice mental health nursing: commercial film as a suggested modality. AB - The purpose of this study was to explore the advantages and disadvantages of using feature films in teaching advanced practice mental health nursing. A phenomenological investigation was conducted of 11 master's student perspectives regarding the use of movies in an on-line course. According to the students, the disadvantages of movies were that films are time consuming to watch and often more dramatic than real-life situations. The advantages of using movies were that films are effective in promoting reflection and enhancing empathy. Students noted that movies poignantly portray emotional issues and are engaging in addition to being a good way both to present educational content and introduce ethical dilemmas. Using movies also may be effective with select clients as a once removed way to introduce therapeutic discussions in outpatient practice. PMID- 12775548 TI - The medical, personal, and social causes of uncertainty in HIV illness. AB - Uncertainty is an important part of the illness experience. Mishel elaborated a theory of uncertainty in acute illness and later expanded the framework to account for uncertainty in chronic illness. Researchers subsequently have investigated the causes and outcomes associated with the uncertainty in illness experience across a variety of medical conditions. The current study applies and extends Mishel's model within the context of HIV illness-related uncertainty. In this qualitative study, focus group methods were used to examine the nature of illness uncertainty experienced by persons living with HIV or AIDS. Findings confirm Mishel's contention that the causes of uncertainty extend beyond those of medical diagnosis, treatment, and recovery to personal and social aspects of daily life. Identified sources of uncertainty may have important mental health and quality of life implications. PMID- 12775549 TI - Use of the Interpersonal Support Evaluation List (ISEL) to guide intervention development with women experiencing abuse. AB - Effective intervention strategies for abused women must be developed that match women's perceived needs. The purpose of this study was to examine the associations among age, race, employment status, education levels, and women's perceptions of social support. Women (n = 40) from 19 to 68 years of age in a family court setting or domestic violence shelter completed the Interpersonal Support Evaluation List (ISEL), which was used to measure subscales of companionship, self-esteem, emotional support, and instrumental support. Abuse was documented by patient reports and protocols of the referring agencies. Significant associations were found between age and self-esteem (r = 0.47, p = 0.002) companionship (r = 0.29, p = 0.07) and emotional support (r = 0.27, p = 0.09). Also significant were the associations between race and self-esteem (r = 0.27, p = 0.10) and employment and instrumental support (r = -0.32, p = 0.08). These significant findings may aid in the development of interventions for abused women that are matched to their social support needs, particularly if the findings are consistent in larger studies. PMID- 12775550 TI - Recovery from intimate partner violence: a difficult journey. AB - In this existential-phenomenological investigation 15 women were interviewed about their experience of recovering from intimate partner violence (IPV). The purpose of the study was to examine the healing process from IPV from the perspective of those who have experienced it. Transcribed interviews were analyzed using a hermeneutic process. The thematic structure of a woman's journey is comprised of three distinct but connected phases of her life: the abusive past, the struggles of freeing herself physically and emotionally from the abuse/past, and finally the healing/growth that occurs as a woman releases herself from the bitterness and anger of the past. Most of the women in this study were able to experience the peace and healing that occurs with recovery, although a few remained emotionally stuck and unable to free themselves from the past. Thus, there are two stories that are told in this study. The first story is about a journey of recovery. Sadly, the second story is about nonrecovery. PMID- 12775551 TI - Patient concerns about seclusion: developing a leaflet. AB - Seclusion continues to be used as a last resort in many acute in-patient mental health facilities, hence, mental health nurses must consider a range of strategies to improve seclusion practice. This article reviews selected literature to glean relevant information to provide to patients regarding seclusion protocols, rationales, and aims. Some postseclusion debriefing, nurse education, and organizational monitoring issues are also briefly discussed. The provision of supplementary written patient information about seclusion processes has the potential to decrease patient anxiety and fear. This initiative involves collaboration with consumer consultants to creatively develop effective solutions to some long-standing patient-identified problems associated with the experience of seclusion. PMID- 12775553 TI - Cholinergic neurotransmission participates in increased food intake induced by NMDA receptor blockade. AB - MK-801, a noncompetitive N-methyl-d-aspartate (NMDA) receptor antagonist, enhances gastric emptying while increasing food intake. Although our previously reported results implicate the vagus in MK-801's effect on feeding, it is not clear whether vagal motor fibers participate in the feeding response. Control of gastric emptying is exerted, in part, by cholinergic vagal motor neurons. Therefore, we examined the ability of MK-801 to increase meal size in the presence or absence of the muscarinic receptor antagonist atropine methyl nitrate. Both central and systemic administration of MK-801 significantly increased intake of 15% sucrose. Intraperitoneal injection of atropine abolished MK-801-induced increase in sucrose intake, whereas administration into the fourth ventricle had no effect. To determine whether augmentation of cholinergic tone produces an enhancement of food intake in the absence of MK-801, we tested the ability of cisapride, a gastric prokinetic agent that promotes acetylcholine release through an action on presynaptic serotonin (5-HT4) receptors, to increase sucrose consumption. Cisapride (500 microg/kg ip) induced a small but significant increase in 15% sucrose intake (15.5 +/- 0.5 ml) compared with NaCl (13.0 +/- 0.6 ml). Furthermore, when MK-801 (100 microg/kg ip) was given in combination with cisapride, intake was significantly higher (19.8 +/- 0.9 ml) than following either agent given alone. Pretreatment with atropine abolished the cisapride induced increase in intake (12.1 +/- 0.9 ml) as well as the increased intake induced by combining MK-801 and cisapride. These results suggest that blockade of NMDA-gated ion channels in the hindbrain increases food intake, in part, via a peripheral muscarinic cholinergic mechanism. PMID- 12775554 TI - Activity of the unique beta-adrenergic Na+/H+ exchanger in trout erythrocytes is controlled by a novel beta3-AR subtype. AB - beta-Adrenoceptors (beta-ARs) are seven-transmembrane domain, G protein-coupled receptors that transduce the cellular effects of epinephrine and norepinephrine and play a pivotal role in the vertebrate stress response. This study reports the cloning and characterization of two previously unreported beta-ARs from the rainbow trout (Oncorhynchus mykiss). Phylogenetic analysis of amino acid sequences indicates that both beta-ARs are homologs of the mammalian beta3-AR. Analysis of tissue expression patterns indicates that one of these trout beta3 adrenoceptors (beta3a-AR) is highly expressed in gill and heart, whereas the second (beta3b-AR) is highly expressed by red blood cells (RBC). Expression of the beta3b-AR in the RBC coupled with the finding of a single category of beta-AR binding sites on RBC membranes provides strong evidence for the control of the trout RBC beta-AR Na+/H+ exchanger (beta-NHE) activity by signaling through this beta3b-subtype and not through a beta1-subtype as previously proposed. The RBC specific trout beta3b-AR exhibits binding characteristics that distinguish this receptor from each of the three pharmacologically defined categories of mammalian beta-ARs (beta1-, beta2-, and beta3-AR). This study is the first to report the presence of a beta3-AR subtype in a fish species, and the proposal that the beta3b-AR controls RBC beta-NHE activity represents a novel role for the beta3-AR subtype in vertebrates. PMID- 12775555 TI - Ontogeny of diet-induced obesity in selectively bred Sprague-Dawley rats. AB - Outbred Sprague-Dawley rats selectively bred for their propensity to develop diet induced obesity (DIO) become heavier on low-fat diet than those bred to be diet resistant (DR) beginning at approximately 5 wk of age. Here we assessed the development of metabolic and neural functions for insights into the origins of their greater weight gain. From week 5 to week 10, chow-fed DIO rats gained 15% more body weight and ate approximately 14% more calories but had only slightly greater adiposity and plasma leptin than DR rats. From day 3 through week 10, DIO and DR rats had similar mRNA expression of arcuate nucleus neuropeptide Y, proopiomelanocortin, agouti-related peptide, and all splice variants of the leptin receptor (OB-R). When fed a high-energy (HE; 31% fat) diet, 7-wk-old DIO rats had a 240% increase in plasma leptin levels after only 3 days. Despite this early leptin rise, they maintained a persistent hyperphagia and became more obese than chow-fed DIO rats and DR rats fed chow or HE diet. Their failure to reduce caloric intake, despite high levels of leptin, suggests that selectively bred DIO rats might have reduced leptin sensitivity similar to that seen in the outbred DIO parent strain. PMID- 12775557 TI - Fetal anemia leads to augmented contractile response to hypoxic stress in adulthood. AB - In response to chronic fetal anemia, coronary blood flow, maximal coronary conductance, and coronary reserve increase. We sought to determine whether chronic fetal anemia alters left ventricular (LV) function in adulthood. We studied adult sheep that had been made anemic for 20 days in utero by phlebotomy. They were transfused just before birth. At 7 mo of age, LV function was measured by pressure-volume loops at rest and during hypoxic stress. The in utero anemia group (n = 8) did not differ from controls (n = 5) with respect to hematocrit, heart and body weight, or baseline hemodynamic parameters. However, the effect of hypoxia (relative to baseline) on multiple indexes of systolic function was different between the two groups. End-systolic elastance increased in the in utero anemia group (baseline to hypoxia) by 4.15 +/- 3.47 mmHg/ml (mean +/- SD) but changed little in controls (0.24 +/- 0.45), which shows that the response to hypoxia was significantly different (P < 0.01) between groups. Similarly, the maximum derivative of LV pressure with respect to time increased in the in utero anemia group (486 +/- 340 mmHg/s,) but on average fell in the controls (-503 +/- 211 mmHg/s) with the response again being significantly different (P < 0.03). We conclude that in sheep, perinatal anemia can alter cardiac responses to hypoxic stress in the adult long after restoration of normocythemia. PMID- 12775556 TI - Effect of AT1 receptor blockade on hepatic redox status in SHR: possible relevance for endothelial function? AB - The study investigated whether the amelioration of endothelial dysfunction by candesartan (2 mg.kg-1.day-1; 10 wk) in spontaneously hypertensive rats (SHR) was associated with modification of hepatic redox system. Systolic arterial pressure (SAP) was higher (P < 0.05) in SHR than in Wistar-Kyoto rats (WKY) and was reduced (P < 0.05) by candesartan in both strains. Acetylcholine (ACh) relaxations were smaller (P < 0.05) and contractions induced by ACh + NG-nitro-l arginine methyl ester (l-NAME) were greater (P < 0.05) in SHR than in WKY. Treatment with candesartan enhanced (P < 0.05) ACh relaxations in SHR and reduced (P < 0.05) ACh + l-NAME contractions in both strains. Expression of aortic endothelial nitric oxide synthase (eNOS) mRNA was similar in WKY and SHR, and candesartan increased (P < 0.05) it in both strains. Aortic mRNA expression of the subunit p22phox of NAD(P)H oxidase was higher (P < 0.05) in SHR than in WKY. Treatment with candesartan reduced (P < 0.05) p22phox expression only in SHR. Malonyl dialdehyde (MDA) levels were higher (P < 0.05), and the ratio reduced/oxidized glutathione (GSH/GSSG) as well as glutathione peroxidase activity (GPx) were lower (P < 0.05) in liver homogenates from SHR than from WKY. Candesartan reduced (P < 0.05) MDA and increased (P < 0.05) GSH/GSSG ratio without affecting GPx. Vessel, lumen, and media areas were bigger (P < 0.05) in SHR than in WKY. Candesartan treatment reduced (P < 0.05) media area in SHR without affecting vessel or lumen area. The results suggest that hypertension is not only associated with elevation of vascular superoxide anions but with alterations of the hepatic redox system, where ANG II is clearly involved. The results further support the key role of ANG II via AT1 receptors for the functional and structural vascular alterations produced by hypertension. PMID- 12775558 TI - A compartmental model of magnesium metabolism in healthy men based on two stable isotope tracers. AB - The aim of this study was to build a compartmental model of magnesium (Mg) kinetics by using data collected from six healthy adult men after oral administration of 26Mg and intravenous administration of 25Mg. Blood, urine, and feces were collected for 12 days after administration of the isotopes. Isotopic ratios were determined by inductively coupled plasma-mass spectrometry. Data were analyzed for each subject using SAAMII. We began with a compartmental model previously proposed (Avioli LV and Berman M. J Appl Physiol 21: 1688-1694, 1966) and developed an alternative approach to resolve the discrepancy between model predicted curves and experimental data. This analysis enables the exploration of 25% of total body Mg that exchanges rapidly from plasma compartment with two extraplasma pools. One of the extraplasma compartments contains 80% of the exchangeable Mg with a transport rate of 48 +/- 13 mg/h. The second exchanges 179 +/- 88 mg of Mg/h. The model permitted estimation of kinetic parameters as well as fractional Mg absorption and fecal endogenous excretion. PMID- 12775559 TI - Period gene expression in mouse endocrine tissues. AB - Circadian rhythms are generated by the oscillating expression of the Per1 and Per2 genes, which are expressed not only in the central brain pacemaker but also in peripheral tissues. Hormones are likely to coordinate physiological function in time. We performed in situ hybridization to localize mPer1 and mPer2 mRNA to particular cell types and tissue compartments in adrenal, thyroid, and testis. BALB/c mice maintained in a 12:12-h light-dark cycle expressed mPer1 in adrenal medulla, particularly in late afternoon and early night. mPer2 mRNA was more intensely expressed in adrenal cortex, especially in afternoon and evening. mPer1 mRNA was detected in thyroid. mPer1 was found in some but not all seminiferous tubules of each mouse at all times of day. Quantitation in C57BL/6 mice revealed a significant increase in the number of heavily labeled seminiferous tubules early in the night. Consistent with in situ hybridization, immunocytochemistry showed PER1 protein in spermatocytes and spermatids (spermatogenic stages VII XII). Staining in spermatogonia and interstitial cells was inconsistent. Double labeling with 5'-bromodeoxyuridine showed PER1 expression first occurring 5 days after DNA replication. We conclude that mPeriod genes are expressed in peripheral endocrine glands. Central regulation, adenohypophyseal control, and functional importance of expression and phase remain to be elucidated. PMID- 12775560 TI - Measurement of aortic input impedance in mice: effects of age on aortic stiffness. AB - Mice are used with increasing frequency as models of human cardiovascular diseases, but significant gaps exist in our knowledge of vascular function in the aging mouse. We determined aortic input impedance spectra, pulse wave velocity, and augmentation index in adult (8-mo-old) and old (29-mo-old) mice to determine whether arterial stiffening occurred with age in mice as it does in humans. Pressure and blood velocity signals measured simultaneously from the same location in the ascending aorta were used to determine input impedance spectra (0 10 harmonics). The first minimum of the impedance modulus occurred at the second harmonic in adult mice but shifted to the fourth harmonic in old mice. Characteristic impedance (average of 2nd-10th harmonic) was 57% higher in old mice: 471 +/- 62 vs. 299 +/- 10 (SE) dyn.s.cm-3 (P < 0.05). Pulse pressure and augmentation index, determined from the aortic pressure signals, were also higher in old mice: 42 +/- 2.2 vs. 29 +/- 4.9 mmHg (P < 0.05) and 37 +/- 5 vs. 14 +/- 2% (P < 0.005). Aortic pulse wave velocity measured from the timing of upstrokes of the Doppler velocity signals was 45% higher in old mice: 416 +/- 22 vs. 286 +/- 14 cm/s (n = 3, P < 0.01). These results reproduce age-related findings reported in humans and confirm that mice may be used as models of age-related vascular stiffening. PMID- 12775561 TI - Oxygen regulation and limitation to cellular respiration in mouse skeletal muscle in vivo. AB - In skeletal muscle, intracellular Po2 can fall to as low as 2-3 mmHg. This study tested whether oxygen regulates cellular respiration in this range of oxygen tensions through direct coupling between phosphorylation potential and intracellular Po2. Oxygen may also behave as a simple substrate in cellular respiration that is near saturating levels over most of the physiological range. A novel optical spectroscopic method was used to measure tissue oxygen consumption (Mo2) and intracellular Po2 using the decline in hemoglobin and myoglobin saturation in the ischemic hindlimb muscle of Swiss-Webster mice. 31P magnetic resonance spectroscopic determinations yielded phosphocreatine concentration ([PCr]) and pH in the same muscle volume. Intracellular Po2 fell to <2 mmHg during the ischemic period without a change in the muscle [PCr] or pH. The constant phosphorylation state despite the decline in intracellular Po2 rejects the hypothesis that direct coupling between these two variables results in a regulatory role for oxygen in cellular respiration. A second set of experiments tested the relationship between intracellular Po2 and Mo2. In vivo Mo2 in mouse skeletal muscle was increased by systemic treatment with 2 and 4 mg/kg body wt 2,4-dinitrophenol to partially uncouple mitochondria. Mo2 was not dependent on intracellular Po2 above 3 mmHg in the three groups despite a threefold increase in Mo2. These results indicate that Mo2 and the phosphorylation state of the cell are independent of intracellular Po2 throughout the physiological range of oxygen tensions. Therefore, we reject a regulatory role for oxygen in cellular respiration and conclude that oxygen acts as a simple substrate for respiration under physiological conditions. PMID- 12775562 TI - Targeted deletion of MMP-2 attenuates early LV rupture and late remodeling after experimental myocardial infarction. AB - Matrix metalloproteinase-2 (MMP-2) is prominently overexpressed both after myocardial infarction (MI) and in heart failure. However, its pathophysiological significance in these conditions is still unclear. We thus examined the effects of targeted deletion of MMP-2 on post-MI left ventricular (LV) remodeling and failure. Anterior MI was produced in 10- to 12-wk-old male MMP-2 knockout (KO) and sibling wild-type (WT) mice by ligating the left coronary artery. By day 28, MI resulted in a significant increase in mortality in association with LV cavity dilatation and dysfunction. The MMP-2 KO mice had a significantly better survival rate than WT mice (56% vs. 85%, P < 0.05), despite a comparable infarct size (50 +/- 3% vs. 51 +/- 3%, P = not significant), heart rate, and arterial blood pressure. The KO mice had a significantly lower incidence of LV rupture (10% vs. 39%, P < 0.05), which occurred within 7 days of MI. The KO mice exerted less LV cavity dilatation and improved fractional shortening after MI by echocardiography. The LV zymographic MMP-2 level significantly increased in WT mice after coronary artery ligation; however, this was completely prevented in KO mice. In contrast, the increase in the LV zymographic MMP-9 level after MI was similar between KO and WT mice. MMP-2 activation is therefore considered to contribute to an early cardiac rupture as well as late LV remodeling after MI. The inhibition of MMP-2 activation may therefore be a potentially useful therapeutic strategy to manage post-MI hearts. PMID- 12775563 TI - Chronic all-trans retinoic acid treatment prevents medial thickening of intramyocardial and intrarenal arteries in spontaneously hypertensive rats. AB - There are in vitro data linking all-trans retinoic acid (atRA) with inhibition of hypertrophy and hyperplasia in cardiomyocytes, vascular smooth muscle cells, and fibroblasts. In the present study, we tested the hypothesis that chronic treatment with atRA may blunt the process of myocardial remodeling in spontaneously hypertensive rats (SHR). Four-week-old male SHR were treated with atRA (5 or 10 mg.kg-1.day-1) given daily for 3 mo by gavage; age- and sex-matched Wistar-Kyoto rats (WKY) and placebo-treated SHR served as controls. At the end of the treatment period, cardiac geometry and function were assessed by Doppler echocardiography. Histological examination and RIA were performed to evaluate medial thickening of intramyocardial and renal arteries, perivascular and interstitial collagen content, and atrial natriuretic peptide (ANP) and IGF-I in the heart, respectively. The novel finding of the present study is that atRA prevented hypertrophy of intramyocardial and intrarenal arteries and ventricular fibrosis. However, atRA treatment did not lower blood pressure or left ventricular weight and left ventricular weight-to-body weight ratio in SHR. atRA did not change cardiac geometry and function as assessed by Doppler echocardiography. atRA showed no influence on either ANP or IGF-I levels. In conclusion, the present study suggests that chronic atRA treatment prevents medial thickening of intramyocardial and intrarenal arteries and ventricular fibrosis during the development of hypertension. Left ventricular hypertrophy and cardiac geometry and function are not changed by atRA treatment. PMID- 12775564 TI - Cardiac retention of [11C]HED in genotyped long QT patients: a potential amplifier role for severity of the disease. AB - Although mutations in cardiac sodium and potassium channel genes are associated with congenital long QT syndrome (LQTS), a "modifier" role of the sympathetic nervous system was proposed to explain the distinct severity of the disease. We evaluated cardiac sympathetic innervation using [11C]hydroxyephedrine ([11C]HED) and positron emission tomography (PET) in genotyped LQTS patients. H215O and [11C]HED PET studies were performed in 11 patients (5 symptomatic) and 8 controls. Perfusion and [11C]HED images were depicted as 36-sector polar maps. Sectorial values of perfusion (H2O%), absolute (HEDRet) and relative retention (HED%Ret) of [11C]HED, and the ratio of HED%Ret to H2O% (HED%Ret/H2O%) were calculated. Normal databases were obtained from controls. Sectorial values below 2SD database values were defined as "outside sectors." Controls and patients showed similar sectorial perfusion. Sectorial HEDRet did not differ between groups, but means of HED%Ret were lower in three sectors for patients (P < 0.05). Three sectors from 3 controls had HED%Ret below 2SD, whereas 36 sectors in 9 patients were outside sectors (P < 0.01). In patients, average HED%Ret/H2O% was lower in 9 sectors (P < 0.05 vs. controls); 2 outside sectors were found in controls, but 43 outside sectors were found in patients (P < 0.01), 77% of them in the 5 symptomatic patients. Heterogeneous [11C]HED retention was localized in the septal, anterior, and lateral walls. Most LQTS patients showed a localized and decreased pattern of [11C]HED retention. The larger number of heterogeneous sectors in symptomatic patients suggests that sympathetic function could play an amplifier role for severity of the disease. PMID- 12775565 TI - Endothelin receptor blockade has an oxygen-saving effect in Dahl salt-sensitive rats with heart failure. AB - The effects of endothelin (ET) receptor blockade on energy utilization in heart failure (HF) are unknown. We administered ET type A (ETA), ET type B (ETB), and ETA/ETB antagonists to isolated hearts from Dahl salt-sensitive (DS) rats with HF and controls. Contractile efficiency was assessed as slope-1 of myocardial O consumption (VO2)-pressure-volume area relation. In HF, ETA and ETA/ETB but not ETB blockade decreased the contractility index (Emax)(-15 +/- 3% and -17 +/- 2%, P < 0.05), excitation-contraction (E-C) coupling VO2 (-39 +/- 4% and -37 +/- 5%, P < 0.01), and efficiency (-15 +/- 4% and -17 +/- 2%, P < 0.05). Despite decreased efficiency, ETA and ETA/ETB blockade decreased total VO2 (-24 +/- 3% and -22 +/- 2%, P < 0.05). Na+/H+ exchanger inhibition decreased Emax and E-C coupling VO2 similar to ETA and ETA/ETB blockade, but did not alter efficiency. In HF, endogenous ET-1 maintains contractility at expense of increased VO2 through ETA receptor activation, likely mediated by Na+/H+ exchange. PMID- 12775566 TI - Transforming growth factor-beta1 down-regulates expression of chemokine stromal cell-derived factor-1: functional consequences in cell migration and adhesion. AB - Chemokine stromal cell-derived factor-1 (SDF-1) is expressed by bone marrow (BM) stromal cells and plays key roles in BM cell migration. Modulation of its expression could affect the migratory capacity of cells trafficking the BM, such as hematopoietic progenitor and leukemic cells. Transforming growth factor-beta1 (TGF-beta1) is present in the BM environment and constitutes a pivotal molecule controlling BM cell proliferation and differentiation. We used the BM stromal cell line MS-5 as a model to investigate whether SDF-1 expression constitutes a target for TGF-beta1 regulation and its functional consequences. We show here that TGF-beta1 down-regulates SDF-1 expression, both at the mRNA level, involving a decrease in transcriptional efficiency, and at the protein level, as detected in lysates and supernatants from MS-5 cells. Reduction of SDF-1 in supernatants from TGF-beta1-treated MS-5 cells correlated with decreased, SDF-1-dependent, chemotactic, and transendothelial migratory responses of the BM model cell lines NCI-H929 and Mo7e compared with their responses to supernatants from untreated MS 5 cells. In addition, supernatants from TGF-beta1-exposed MS-5 cells had substantially lower efficiency in promoting integrin alpha4beta1-mediated adhesion of NCI-H929 and Mo7e cells to soluble vascular cell adhesion molecule-1 (sVCAM-1) and CS-1/fibronectin than their untreated counterparts. Moreover, human cord blood CD34+ hematopoietic progenitor cells displayed SDF-1-dependent reduced responses in chemotaxis, transendothelial migration, and up-regulation of adhesion to sVCAM-1 when supernatants from TGF-beta1-treated MS-5 cells were used compared with supernatants from untreated cells. These data indicate that TGF beta1-controlled reduction in SDF-1 expression influences BM cell migration and adhesion, which could affect the motility of cells trafficking the bone marrow. PMID- 12775567 TI - Role of macrophages in the generation of circulating blood nucleosomes from dead and dying cells. AB - After apoptosis or necrosis, macrophages clear dead cells by phagocytosis. Although this process is efficient, circulating nucleosomes can occur in certain diseases, presumably reflecting either increased production or impaired clearance. To investigate the generation of blood nucleosomes, graded numbers of apoptotic and necrotic cells were administered to healthy mice, and levels of blood nucleosomes and DNA were determined. Using Jurkat cells as a model, nucleosomes and DNA were detected in the blood after the administration of 108 apoptotic or necrotic cells per mouse by the intraperitoneal route. The kinetics of the response were similar for both types of cells. The role of macrophages was assessed by eliminating these cells with clodronate liposomes or silica. Although clodronate treatment alone produced a peak level of blood DNA, the subsequent administration of dead cells caused no change in DNA levels. In contrast, silica treatment alone did not elicit a blood DNA response, though this treatment limited the rise in DNA from administered cells. Molecular studies showed that the blood DNA following the administration of apoptotic or necrotic cells arose from the mouse and the Jurkat cells, and its size distribution was consistent with apoptosis. Together, these findings suggest that the generation of blood nucleosomes depends on macrophages, with apoptosis a concomitant of a high burden of dead and dying cells. PMID- 12775568 TI - Intronic BCL-6 mutations are preferentially targeted to the translocated allele in t(3;14)(q27;q32) non-Hodgkin B-cell lymphoma. AB - Translocations and somatic mutations are common genetic alterations of the BCL-6 gene on chromosome 3q27 in B-cell lymphoma, with implications for lymphomagenesis. The 2 events may have linked origins and can influence juxtaposed loci. To evaluate this further, we compared mutations occurring within the major mutation cluster region of the translocated and untranslocated BCL-6 alleles in 7 t(3;14)(q27;14q32) lymphomas. In 6 of 7 cases, the translocated allele revealed significantly higher mutations (mean, 5.8 x 10-2 bp-1) than did the untranslocated allele (mean, 5.3 x 10-3 bp-1; P <.01). The increase mapped to der(14q32), which retains the BCL-6 promoter and is transcriptionally active, as revealed by fusion transcripts and ongoing somatic mutations, absent in the der(3q27) region. These results indicate that enhanced mutational activity at the translocated allele may be a consequence of loss of cis regulatory elements or gain of IgH enhancer elements. Junctional sequences indicate translocation origins from earlier BCL-6 mutations and switch recombinase events. PMID- 12775569 TI - Distribution of marrow repopulating cells between bone marrow and spleen early after transplantation. AB - Whether hematopoietic stem cells (HSCs) home selectively to bone marrow (BM) early after transplantation remains an issue of debate. Better understanding of homing mechanisms may benefit BM transplantation protocols in cases of limited graft cell number or nonmyeloablative conditioning regimens. Using flow cytometry and serial transplantation to stringently identify HSCs, trafficking patterns of long-term engrafting cells were mapped between BM and spleen early after transplantation. Low-density BM cells were tracked in irradiated or nonirradiated mice 1, 3, 6, and 20 hours after transplantation, at which time recipient BM and spleen were analyzed for recovery of primitive donor cells by phenotype and adhesion molecule expression. In addition, phenotypically defined HSC-enriched or HSC-depleted grafts were tracked 20 hours after transplantation in recipient BM and spleen and analyzed for recovery and long-term repopulating potential in mice undergoing serial transplantation. Regardless of irradiation status, recovery of donor Sca-1+ lin- cells was higher at most time points in recipient BM than in spleen, while recovery of total Sca-1+ cells was variable. A significantly higher percentage of BM-homed donor Sca-1+ cells expressed CD43, CD49e, and CD49d 20 hours after transplantation than spleen-homed cells, which contained significantly more non-HSC phenotypes. Furthermore, BM-homed cells were significantly enriched for cells capable of secondary multilineage hematopoiesis in mice undergoing serial transplantation compared with spleen-homed cells. These results support the notion of specific homing of HSCs to BM by 20 hours after transplantation and provide a basis for the enhanced engraftment potential afforded some Sca-1+ lin- cells subfractionated on the basis of adhesion molecule expression. PMID- 12775570 TI - MHC class II signal transduction in human dendritic cells induced by a natural ligand, the LAG-3 protein (CD223). AB - On encountering a danger signal, dendritic cells (DCs) undergo a complex maturation process and become specialized in antigen presentation. We previously reported that engagement of major histocompatibility complex (MHC) class II molecules located on immature DCs in membrane rafts by lymphocyte activation gene 3 (LAG-3; CD223) leads to DC maturation. In contrast, exposure of DCs to class II specific monoclonal antibodies (mAbs) did not lead to maturation. Here, we have investigated the signal transduction pathways involved in the LAG-3-induced maturation of human monocyte-derived DCs. We first show that areas of raft aggregation (both cholesterol rich and CDw78 microdomains) could be visualized using a soluble LAG-3 protein and confocal microscopy. Engagement of class II molecules by both its natural ligand LAG-3 and class II mAb induces rapid protein phosphorylation of phospholipase Cgamma2 (PLCgamma2) and p72syk as well as activation of phosphatidyl inositol 3-kinase/Akt, p42/44 extracellular signal regulated protein kinase, and p38 mitogen-activated protein kinase pathways. Studies using inhibitors demonstrate that these 3 pathways are all important in inducing the maturation process of LAG-3-stimulated DCs. When class II molecules were ligated with LAG-3 versus specific antibody, differences in the phosphorylation pattern of c-Akt were observed. Thus, MHC class II signaling in DCs involves several pathways that have to be finely regulated to lead to cell activation and maturation. PMID- 12775571 TI - A mathematical model of the contribution of endothelial progenitor cells to angiogenesis in tumors: implications for antiangiogenic therapy. AB - The traditional view of angiogenesis emphasizes proliferation and migration of vessel wall-associated endothelial cells. However, circulating endothelial progenitor cells have recently been shown to contribute to tumor angiogenesis. Here we quantify the relative contributions of endothelial and endothelial progenitor cells to angiogenesis using a mathematical model. The model predicts that during the early stages of tumor growth, endothelial progenitors have a significant impact on tumor growth and angiogenesis, mediated primarily by their localization in the tumor, not by their proliferation. The model also shows that, as the tumor grows, endothelial progenitors adhere preferentially near the tumor periphery, coincident with the location of highest vascular density, supporting their potential utility as vectors for targeted delivery of therapeutics. Model simulations of various antiangiogenic strategies show that those therapies that effectively target both endothelial and endothelial progenitor cells, either by restoring the balance between angiogenic stimulators and inhibitors or by targeting both types of cells directly, are most effective at delaying tumor growth. The combination of continuous low-dose chemotherapy and antiangiogenic therapy is predicted to have the most significant effect on therapeutic outcome. The model offers new insight into tumor angiogenesis with implications for the rational design of antiangiogenic therapy. PMID- 12775572 TI - Anergic T cells exert antigen-independent inhibition of cell-cell interactions via chemokine metabolism. AB - Due to their ability to inhibit antigen-induced T-cell activation in vitro and in vivo, anergic T cells can be considered part of the spectrum of immunoregulatory T lymphocytes. Here we report that both murine and human anergic T cells can impair the ability of parenchymal cells (including endothelial and epithelial cells) to establish cell-cell interactions necessary to sustain leukocyte migration in vitro and tissue infiltration in vivo. The inhibition is reversible and cell-contact dependent but does not require cognate recognition of the parenchymal cells to occur. Instrumental to this effect is the increased cell surface expression and enzymatic activity of molecules such as CD26 (dipeptidyl peptidase IV), which may act by metabolizing chemoattractants bound to the endothelial/epithelial cell surface. These results describe a previously unknown antigen-independent anti-inflammatory activity by locally generated anergic T cells and define a novel mechanism for the long-known immunoregulatory properties of these cells. PMID- 12775573 TI - Anti-third-party veto CTLs overcome rejection of hematopoietic allografts: synergism with rapamycin and BM cell dose. AB - Several bone marrow cells and lymphocyte subpopulations, known as "veto cells," were shown to induce transplantation tolerance across major histocompatibility antigens. Some of the most potent veto cells are of T-cell origin, and in particular a very strong veto activity was documented for cytotoxic T-lymphocyte (CTL) lines or clones. However, these cells also possess marked graft-versus-host (GVH) reactivity. In the present study we evaluated a new approach to deplete CTLs of antihost clones by stimulating the donor T cells against third-party stimulators in the absence of exogenous interleukin 2 (IL-2). We demonstrate that such CTLs are depleted of GVH reactivity while maintaining marked veto activity in vitro. Furthermore, marked synergism was exhibited between the veto CTLs and rapamycin when tested in a murine model, which measures T-cell-mediated bone marrow allograft rejection, or in sublethally irradiated allogeneic hosts. Our results suggest that engraftment of early progenitors could be enhanced by using host-nonreactive anti-third-party CTLs, in conjunction with nonmyeloablative rapamycin-based conditioning protocols, thereby significantly reducing the toxicity of allogeneic transplantation. PMID- 12775574 TI - Induction of allopeptide-specific human CD4+CD25+ regulatory T cells ex vivo. AB - Although CD4+CD25+ regulatory T cells are pivotal in the prevention of autoimmunity and appear to mediate transplantation tolerance, little is known concerning their antigen specificity. Here we describe the induction of a human CD4+CD25+ regulatory T-cell line specific for a defined peptide alloantigen (human leukocyte antigen A2 [HLA-A2] 138-170) by priming purified CD4+CD25+ cells ex vivo. The regulatory cells were anergic and retained their ability to suppress antigen-driven responses of CD4+CD25- cells. They inhibited not only interleukin 2 (IL-2) secretion by CD4+CD25- T cells specific for the same peptide but also direct alloresponse of naive CD4+CD25- T cells stimulated by semiallogeneic dendritic cells (DCs) in the presence of the peptide ("linked suppression"). They also suppressed the response of CD4+ T cells specific for viral and bacterial antigens. The suppressive T-cell line showed sustained high CD25 expression. These findings suggest that peripheral CD4+CD25+ regulatory cells are a precommitted cell lineage from which cells with specificity for non-self-peptides can be selected. This may pave the way for inducing and expanding peptide antigen specific regulatory T cells ex vivo for cell therapy in transplantation, allergy, and autoimmune disease. PMID- 12775575 TI - von Willebrand factor but not alpha-thrombin binding to platelet glycoprotein Ibalpha is influenced by the HPA-2 polymorphism. AB - OBJECTIVE: Glycoprotein (GP) Ibalpha is the functionally dominant subunit of the platelet GPIb-IX-V receptor complex. The N-terminal domain of the GPIbalpha chain contains binding sites for alpha-thrombin and von Willebrand factor (VWF). The human platelet alloantigen (HPA)-2 polymorphism of the GPIbalpha gene is associated with a C/T transition at nucleotide 1018, resulting in a Thr/Met dimorphism at residue 145 of GPIbalpha. To study the structural and functional effects of this dimorphism, N-terminal fragments (AA1-289) of the HPA-2a and HPA 2b alloform of GPIbalpha expressed in CHO cells were used. METHODS AND RESULTS: Of 74 moAbs directed against human GPIbalpha, 2 antibodies with epitope between AA1-59 could differentiate between both alloforms. In addition, VWF bound with a higher affinity to the recombinant HPA-2a fragment or to homozygous HPA-2a platelets. In contrast, no difference was found in the binding of alpha-thrombin to the recombinant alloform fragments or of antibodies directed against the alpha thrombin binding anionic sulfated tyrosine sequence (AA269-282). CONCLUSIONS: Whereas the Thr145Met dimorphism does not affect alpha-thrombin binding, it does influence the conformation of the N-terminal flanking region and first leucine rich repeat of GPIbalpha and by this has an effect on VWF binding. PMID- 12775576 TI - Specific phospholipid oxidation products inhibit ligand activation of toll-like receptors 4 and 2. AB - OBJECTIVE: We have previously shown that phospholipid oxidation products of 1 palmitoyl-2-arachidonoyl-sn-glycero-3-phosphorylcholine (ox-PAPC) inhibit lipopolysaccharide (LPS)-induced E-selectin expression and neutrophil binding in human aortic endothelial cells (HAECs). The current studies identify specific phospholipids that inhibit chemokine induction by Toll-like receptor-4 (TLR4) and -2 (TLR2) ligands inECs and macrophages. METHODS AND RESULTS: Measurements of interleukin (IL)-8 and monocyte chemotactic protein-1 levels secreted from ox PAPC- and LPS-cotreated ECs indicate that ox-PAPC inhibits activation of TLR4 by LPS. The effects of IL-1beta and tumor necrosis factor-alpha, which utilize the same intracellular signaling molecules, were not inhibited. Cell fractionation and immunofluorescence analyses demonstrate that LPS induces membrane translocation of the LPS receptor complex to a lipid raft/caveolar fraction in ECs. Ox-PAPC inhibits this translocation and alters caveolin-1 distribution. Supporting an important role for caveolae in LPS action, overexpression of caveolin-1 enhanced LPS-induced IL-8 synthesis. Ox-PAPC also inhibits the effect of TLR2 and TLR4 ligands in human macrophages. CONCLUSIONS: These studies report a novel mechanism that involves alterations to lipid raft/caveolar processing, by which specific phospholipid oxidation products inhibit activation by TLR4 and TLR2 ligands. These studies have broader implications for the role of ox-PAPC as a regulator of specific lipid raft/caveolar function. PMID- 12775577 TI - Genetic loci contribute to the progression of vascular and cardiac hypertrophy in salt-sensitive spontaneous hypertension. AB - OBJECTIVE: The salt-sensitive Dahl rat and the spontaneously hypertensive rat develop comparable spontaneous hypertension on a low-salt diet, whereas only the salt-sensitive Dahl rat strain develops a striking increase in blood pressure and cardiovascular hypertrophy on a high-salt diet. We set out to identify quantitative trait loci (QTLs) contributing to the progression of salt-induced organ damage in hypertension by studying an F2 population derived from both strains. METHODS AND RESULTS: We determined systolic blood pressure (SBP), vascular aortic hypertrophy (AH), cardiac left ventricular (LV) hypertrophy (LVH), and LV fibrosis in 230 male F2-animals on a high-salt diet. A strong correlation between AH and LVH was found (r=0.58, P<0.0001), and genome-wide QTL mapping detected suggestive or significant QTLs in overlapping chromosomal fragments for AH and LVH on chromosomes 1, 3, and 19, respectively. A significant influence of SBP on the extent of LVH and AH was evident at all QTLs, although significant linkage to SBP (together with LVH) was only found on chromosome 9. No QTLs for LV fibrosis were detected. CONCLUSIONS: This study demonstrates a strong correlation between AH and LVH in salt-sensitive hypertension and identifies QTLs contributing to the progression of cardiovascular hypertrophy in this condition. PMID- 12775578 TI - C-reactive protein, interleukin-6, and fibrinogen as predictors of coronary heart disease: the PRIME Study. AB - OBJECTIVE: This study was undertaken to examine the association of plasma inflammatory markers such as C-reactive protein (CRP), interleukin-6, and fibrinogen with the incidence of coronary heart disease within the prospective cohort study on myocardial infarction (PRIME study). METHODS AND RESULTS: Multiple risk factors were recorded at baseline in 9758 men aged 50 to 59 years who were free of coronary heart disease (CHD) on entry. Nested case-control comparisons were carried out on 317 participants who suffered myocardial infarction (MI)-coronary death (n=163) or angina (n=158) as an initial CHD event during a follow-up for 5 years. After adjustment for traditional risk factors, incident MI-coronary death, but not angina, was significantly associated with CRP, interleukin-6, and fibrinogen, but only interleukin-6 remained significantly associated with MI-coronary death when the 3 inflammatory markers were included in the model. The different interleukin-6 levels in Northern Ireland and France partly explained the difference in risk between these countries. Interleukin-6 appeared as a risk marker of MI-coronary death, and it improved the definition of CHD risk beyond LDL cholesterol. CONCLUSIONS: This association may reflect the underlying inflammatory reaction located in the atherosclerotic plaque or a genetic susceptibility on the part of CHD subjects to answer a proinflammatory stimulus and subsequent increase in hepatic CRP gene expression. PMID- 12775579 TI - High-density lipoprotein stimulates endothelial cell migration and survival through sphingosine 1-phosphate and its receptors. AB - OBJECTIVE: Plasma high-density lipoprotein (HDL) level is inversely correlated with the risk of atherosclerosis. However, the cellular mechanism by which HDL exerts antiatherogenic actions is not well understood. In this study, we focus on the lipid components of HDL as mediators of the lipoprotein-induced antiatherogenic actions. METHODS AND RESULTS: HDL and sphingosine 1-phosphate (S1P) stimulated the migration and survival of human umbilical vein endothelial cells. These responses to HDL and S1P were almost completely inhibited by pertussis toxin and other specific inhibitors for intracellular signaling pathways, although the inhibition profiles of migration and survival were different. The HDL-stimulated migration and survival of the cells were markedly inhibited by antisense oligonucleotides against the S1P receptors EDG-1/S1P1 and EDG-3/S1P3. Cell migration was sensitive to both receptors, but cell survival was exclusively sensitive to S1P1. The S1P-rich fraction and chromatographically purified S1P from HDL stimulated cell migration, but the rest of the fraction did not, as was the case of the cell survival. CONCLUSIONS: HDL-induced endothelial cell migration and survival may be mediated by the lipoprotein component S1P and the lipid receptors S1P1 and S1P3. PMID- 12775580 TI - Rho-ROCK-LIMK-cofilin pathway regulates shear stress activation of sterol regulatory element binding proteins. AB - Previous studies have shown that integrin activation and fluid shear stress can modulate the activity of sterol regulatory element binding proteins (SREBPs) in vascular endothelial cells. We investigated the role of small GTPase Rho-mediated signal transduction pathway in this mode of SREBP activation. Fluid shear stress activates the Rho downstream effectors ROCK, LIM kinase (LIMK), and cofilin. The various negative mutants of RhoA, ROCK, LIMK, and cofilin can block the shear stress activation of SREBPs. The shear stress-activated SREBP depends on S2P proteases but not caspase-3. Mechanistically, the endoplasmic reticulum-to-Golgi transport of SREBP cleavage-activating protein requires the actin-based cytoskeleton and is enhanced by the Rho-ROCK-LIMK-cofilin pathway. By enhancing the SREBP-mediated cholesterol metabolism, this unique mechanism may contribute to endothelial cell functions under flow. PMID- 12775581 TI - Immunomediated and ischemia-independent inflammation of coronary microvessels in unstable angina. AB - This study investigated whether the myocardium is involved in the acute inflammatory reaction associated with bursts of unstable angina (UA). We looked for the presence of activated DR+ inflammatory cells and the expression patterns, localization, and immunostaining identification of genes for cytokines (IL-1beta, TNF-alpha, IL-6, and IFN-gamma), MCP-1, and iNOS in the left ventricle biopsies from 2-vessel disease anginal patients, 24 with UA and 12 with stable angina (SA), who underwent coronary bypass surgery. Biopsy specimens from 6 patients with mitral stenosis who underwent valve replacement were examined as control hearts (CHs). Plasma levels of IL-2 soluble receptor (sIL-2R) were measured as a marker of systemic immune reaction. In CHs, DR+ cells were undetectable, and cytokine and iNOS mRNA expression were negligible. UA patients had higher sIL-2R levels than SA patients (P<0.01), and their biopsy specimens showed both numerous DR+ cells identified as lymphocytes, macrophages, endothelial cells, and elevated expression levels of cytokine and iNOS genes (from 2.4- to 6.1-fold vs SA; P<0.01). Cytokine and iNOS genes and proteins were localized in endothelial cells without involvement of myocytes. IL-1beta and MCP-1 mRNAs were nearly undetectable. No significant differences were found in the number of DR+ cells, levels of cytokine, and iNOS genes between potentially ischemic and nonischemic left ventricle areas. In SA specimens, DR+ cells were very rare and only mRNAs for TNF-alpha and iNOS genes were overexpressed versus CHs. These results indicated that an acute immunomediated inflammatory reaction, essentially involving coronary microvessels, is demonstrable in UA patients. PMID- 12775582 TI - Acute inhibition of myoglobin impairs contractility and energy state of iNOS overexpressing hearts. AB - Elevated cardiac levels of nitric oxide (NO) generated by inducible nitric oxide synthase (iNOS) have been implicated in the development of heart failure. The surprisingly benign phenotype of recently generated mice with cardiac-specific iNOS overexpression (TGiNOS) provided the rationale to investigate whether NO scavenging by oxymyoglobin (MbO2) yielding nitrate and metmyoglobin (metMb) is involved in preservation of myocardial function in TGiNOS mice. 1H nuclear magnetic resonance (NMR) spectroscopy was used to monitor changes of cardiac myoglobin (Mb) metabolism in isolated hearts of wild-type (WT) and TGiNOS mice. NO formation by iNOS resulted in a significant decrease of the MbO2 signal and a concomitantly emerging metMb signal in spectra of TGiNOS hearts only (DeltaMbO2: 46.3+/-38.4 micromol/kg, DeltametMb: +41.4+/-17.6 micromol/kg, n=6; P<0.05) leaving contractility and energetics unaffected. Inhibition of the Mb-mediated NO degradation by carbon monoxide (20%) led to a deterioration of myocardial contractility in TGiNOS hearts (left ventricular developed pressure: 78.2+/-8.2% versus 96.7+/-4.6% of baseline, n=6; P<0.005), which was associated with a profound pertubation of cardiac energy state as assessed by 31P NMR spectroscopy (eg, phosphocreatine: 13.3+/-1.3 mmol/L (TGiNOS) versus 15.9+/-0.7 mmol/L (WT), n=6; P<0.005). These alterations could be fully antagonized by the NOS inhibitor S-ethylisothiourea. Our findings demonstrate that myoglobin serves as an important cytoplasmic buffer of iNOS-derived NO, which determines the functional consequences of iNOS overexpression. PMID- 12775583 TI - Dismantling of cadherin-mediated cell-cell contacts modulates smooth muscle cell proliferation. AB - Proliferation of vascular smooth muscle cells (VSMCs) contributes to intimal thickening during atherosclerosis and restenosis. The cadherins are transmembrane proteins, which form cell-cell contacts and may regulate VSMC proliferation. In this study, N-cadherin protein concentration was significantly reduced by stimulation of proliferation with fetal calf serum (FCS) and platelet-derived growth factor-BB (PDGF-BB) in human saphenous vein VSMCs. Furthermore, overexpression of a truncated N-cadherin, which acts as a dominant-negative increased VSMC proliferation. The amount of an extracellular fragment of N cadherin (approximately 90 kDa) in the media after 24 hours was increased by 12 fold by FCS and 11-fold by PDGF-BB, suggesting that N-cadherin levels are regulated by proteolytic shedding. Incubation with a synthetic metalloproteinase inhibitor or adenoviral overexpression of the endogenous tissue inhibitors of metalloproteinases (TIMPs) demonstrated that metalloproteinase activity was responsible in part for this proteolysis. Although total levels of beta-catenin protein were not affected, beta-catenin was translocated to the nucleus after stimulation with FCS and PDGF-BB. Our data indicates cadherin-mediated cell-cell contacts modulate proliferation in VSMCs. Furthermore, disruption of N-cadherin cell-cell contacts mediated in part by metalloproteinase activity occurs during VSMC proliferation, releasing beta-catenin and possibly inducing beta-catenin mediated intracellular signaling. PMID- 12775584 TI - EphA4-mediated Rho activation via Vsm-RhoGEF expressed specifically in vascular smooth muscle cells. AB - Rho-kinase, an effector of Rho GTPase, increases the contractility of vascular smooth muscle by phosphorylating myosin light chain (MLC) and by inactivating MLC phosphatase. A wide variety of extracellular stimuli activate RhoA via G protein coupled receptors. In the present study, we demonstrate a novel cell-cell interaction-mediated Rho activation signaling pathway in vascular smooth muscle cells (VSMCs). Among many receptor tyrosine kinases, the Eph family receptors are unique in that they require cell-cell interaction to engage their ligands, ephrin. We found that a novel VSMC-specific guanine nucleotide exchange factor (GEF) for Rho (Vsm-RhoGEF/KIAA0915) was expressed specifically in VSMCs of several organs including the heart, aorta, liver, kidney, and spleen, as examined by the immunohistochemical analysis using a specific antibody against Vsm-RhoGEF. Based on the association of Vsm-RhoGEF with EphA4 in quiescent cells, we tested whether EphA4 and Vsm-RhoGEF were expressed in the same tissue and further studied the molecular mechanism of Vsm-RhoGEF regulation by EphA4. Immunohistochemical analysis showed that EphA4 and Vsm-RhoGEF expression overlapped in VSMCs. Additionally, tyrosine phosphorylation of Vsm-RhoGEF induced by EphA4 upon ephrin-A1 stimulation enhanced the Vsm-RhoGEF activity for RhoA. The requirement of Vsm-RhoGEF for ephrin-A1-induced assembly of actin stress fibers in VSMCs was shown by the overexpression of a dominant-negative form of VSM-RhoGEF and by the depletion of Vsm-RhoGEF using RNA interference. These results suggested that ephrin-A1-triggered EphA4-Vsm-RhoGEF-RhoA pathway is involved in the cell-cell interaction-mediated RhoA activation that regulates vascular smooth muscle contractility. PMID- 12775585 TI - Hemodynamics is a key epigenetic factor in development of the cardiac conduction system. AB - The His-Purkinje system (HPS) is a network of conduction cells responsible for coordinating the contraction of the ventricles. Earlier studies using bipolar electrodes indicated that the functional maturation of the HPS in the chick embryo is marked by a topological shift in the sequence of activation of the ventricle. Namely, at around the completion of septation, an immature base-to apex sequence of ventricular activation was reported to convert to the apex-to base pattern characteristic of the mature heart. Previously, we have proposed that hemodynamics and/or mechanical conditioning may be key epigenetic factors in development of the HPS. We thus hypothesized that the timing of the topological shift marking maturation of the conduction system is sensitive to variation in hemodynamic load. Spatiotemporal patterns of ventricular activation (as revealed by high-speed imaging of fluorescent voltage-sensitive dye) were mapped in chick hearts over normal development, and following procedures previously characterized as causing increased (conotruncal banding, CTB) or reduced (left atrial ligation, LAL) hemodynamic loading of the embryonic heart. The results revealed that the timing of the shift to mature activation displays striking plasticity. CTB led to precocious emergence of mature HPS function relative to controls whereas LAL was associated with delayed conversion to apical initiation. The results from our study indicate a critical role for biophysical factors in differentiation of specialized cardiac tissues and provide the basis of a new model for studies of the molecular mechanisms involved in induction and patterning of the HPS in vivo. PMID- 12775586 TI - Role of the cytosolic chaperones Hsp70 and Hsp90 in maturation of the cardiac potassium channel HERG. AB - The human ether-a-gogo-related gene (hERG) encodes the alpha subunit of the cardiac potassium current IKr. Several mutations in hERG produce trafficking deficient channels that may cause hereditary long-QT syndrome and sudden cardiac death. Although hERG currents have been studied extensively, little is known about the proteins involved in maturation and trafficking of hERG. Using immunoprecipitations, we show that the cytosolic chaperones heat shock protein (Hsp) 70 and Hsp90, but not Grp94, interact with hERG wild type (WT) during maturation. The specific Hsp90 inhibitor geldanamycin prevents maturation and increases proteasomal degradation of hERG WT, while reducing hERG currents in heterologous expression systems. In ventricular myocytes, inhibition of Hsp90 also decreases IKr, whereas geldanamycin had no effect on IKs or heterologously expressed Kv2.1 and Kv1.5 currents. Both Hsp90 and Hsp70 interact directly with the core-glycosylated form of hERG WT present in the endoplasmic reticulum but not the fully glycosylated, cell-surface form. For the trafficking-deficient LQT2 mutants, hERG R752W and hERG G601S, interactions with Hsp90 and Hsp70 are increased as both mutants remained tightly associated with Hsp90 and Hsp70 in the endoplasmic reticulum. Incubation at lower temperature for R752W or with the hERG blocker astemizole for G601S dissociates channel-chaperone complexes and restores trafficking. In contrast, nonfunctional but trafficking-competent hERG G628S is released from chaperone complexes during maturation comparable to WT. We conclude that Hsp90 and Hsp70 are crucial for the maturation of hERG WT as well as the retention of trafficking-deficient LQT2 mutants. The full text of this article is available online at http://www.circresaha.org. PMID- 12775588 TI - Information from drug companies and opinion leaders. PMID- 12775587 TI - No more free lunches. PMID- 12775589 TI - Haemophilus influenzae type b vaccine--booster campaign. PMID- 12775590 TI - Human tissue engineered products--drugs or devices? PMID- 12775593 TI - Toronto succumbs to SARS a second time. PMID- 12775594 TI - WHO to push for swift implementation of tobacco accord. PMID- 12775595 TI - Drug company sponsorship of education could be replaced at a fraction of its cost. PMID- 12775597 TI - Surgeon suspended in transplant row. PMID- 12775599 TI - Surgeon calls for legalisation of payment to kidney donors. PMID- 12775601 TI - Only half of Dutch doctors report euthanasia, report says. PMID- 12775603 TI - Consultants threaten strike over contract stalemate. PMID- 12775604 TI - World Medical Association reviews doctors' links with drug companies. PMID- 12775605 TI - Hundreds die in northern Iraq from land mines and unexploded munitions. PMID- 12775607 TI - Urologist cleared of manslaughter. PMID- 12775608 TI - War veterans lose compensation battle. PMID- 12775609 TI - New WHO chief pledges to expand network to stem disease outbreaks. PMID- 12775610 TI - NICE recommends new treatment for breast and bowel cancer. PMID- 12775611 TI - WHO continues fight against SARS. PMID- 12775612 TI - Data show that SARS is gradually coming under control. PMID- 12775614 TI - Pharmaceutical industry sponsorship and research outcome and quality: systematic review. AB - OBJECTIVE: To investigate whether funding of drug studies by the pharmaceutical industry is associated with outcomes that are favourable to the funder and whether the methods of trials funded by pharmaceutical companies differ from the methods in trials with other sources of support. METHODS: Medline (January 1966 to December 2002) and Embase (January 1980 to December 2002) searches were supplemented with material identified in the references and in the authors' personal files. Data were independently abstracted by three of the authors and disagreements were resolved by consensus. RESULTS: 30 studies were included. Research funded by drug companies was less likely to be published than research funded by other sources. Studies sponsored by pharmaceutical companies were more likely to have outcomes favouring the sponsor than were studies with other sponsors (odds ratio 4.05; 95% confidence interval 2.98 to 5.51; 18 comparisons). None of the 13 studies that analysed methods reported that studies funded by industry was of poorer quality. CONCLUSION: Systematic bias favours products which are made by the company funding the research. Explanations include the selection of an inappropriate comparator to the product being investigated and publication bias. PMID- 12775616 TI - Uptake of HIV screening in genitourinary medicine after change to "opt-out" consent. PMID- 12775615 TI - Evidence b(i)ased medicine--selective reporting from studies sponsored by pharmaceutical industry: review of studies in new drug applications. AB - OBJECTIVES: To investigate the relative impact on publication bias caused by multiple publication, selective publication, and selective reporting in studies sponsored by pharmaceutical companies. DESIGN: 42 placebo controlled studies of five selective serotonin reuptake inhibitors submitted to the Swedish drug regulatory authority as a basis for marketing approval for treating major depression were compared with the studies actually published (between 1983 and 1999). RESULTS: Multiple publication: 21 studies contributed to at least two publications each, and three studies contributed to five publications. Selective publication: studies showing significant effects of drug were published as stand alone publications more often than studies with non-significant results. Selective reporting: many publications ignored the results of intention to treat analyses and reported the more favourable per protocol analyses only. CONCLUSIONS: The degree of multiple publication, selective publication, and selective reporting differed between products. Thus, any attempt to recommend a specific selective serotonin reuptake inhibitor from the publicly available data only is likely to be based on biased evidence. PMID- 12775617 TI - Systematic review of the effectiveness of stage based interventions to promote smoking cessation. AB - OBJECTIVE: To evaluate the effectiveness of interventions using a stage based approach in bringing about positive changes in smoking behaviour. DESIGN: Systematic review. DATA SOURCES: 35 electronic databases, catalogues, and internet resources (from inception to July 2002). Bibliographies of retrieved references were scanned for other relevant publications, and authors were contacted if necessary. RESULTS: 23 randomised controlled trials were reviewed; two reported details of an economic evaluation. Eight trials reported effects in favour of stage based interventions, three trials showed mixed results, and 12 trials found no statistically significant differences between a stage based intervention and a non-stage based intervention or no intervention. Eleven trials compared a stage based intervention with a non-stage based intervention, and one reported statistically significant effects in favour of the stage based intervention. Two studies reported mixed effects, and eight trials reported no statistically significant differences between groups. The methodological quality of the trials was mixed, and few reported any validation of the instrument used to assess participants' stage of change. Overall, the evidence suggests that stage based interventions are no more effective than non-stage based interventions or no intervention in changing smoking behaviour. CONCLUSIONS: Limited evidence exists for the effectiveness of stage based interventions in changing smoking behaviour. PMID- 12775618 TI - Characteristics of general practitioners who frequently see drug industry representatives: national cross sectional study. PMID- 12775619 TI - Diagnosis, investigation, and management of deep vein thrombosis. PMID- 12775620 TI - Chronic stable angina: treatment options. PMID- 12775621 TI - Who pays for the pizza? Redefining the relationships between doctors and drug companies. 1: entanglement. PMID- 12775622 TI - Who pays for the pizza? Redefining the relationships between doctors and drug companies. 2: Disentanglement. PMID- 12775623 TI - How to dance with porcupines: rules and guidelines on doctors' relations with drug companies. PMID- 12775624 TI - How can research ethics committees protect patients better? PMID- 12775625 TI - Medical journals and pharmaceutical companies: uneasy bedfellows. PMID- 12775626 TI - Unhealthy spin. PMID- 12775627 TI - Relationships between the pharmaceutical industry and patients' organisations. PMID- 12775628 TI - Journals should select drug advertisements more carefully. PMID- 12775629 TI - British Thoracic Society corrects damaging impression. PMID- 12775630 TI - Charities and patient groups should declare interests. PMID- 12775632 TI - Shared scheme for assessing drugs for multiple sclerosis: dealing with uncertainties about cost effectiveness of treatments is difficult problem. PMID- 12775633 TI - Shared scheme for assessing drugs for multiple sclerosis: cost effective provision of effective treatments for multiple sclerosis. PMID- 12775634 TI - Shared scheme for assessing drugs for multiple sclerosis: why are eyes tightly shut to considering causes other than autoimmunity? PMID- 12775635 TI - Doctors and managers: successful partnerships in New Zealand have been incorrectly portrayed. PMID- 12775636 TI - Doctors and managers: capacity and funding need to be increased. PMID- 12775637 TI - Doctors and managers: NHS is unmanageable corporate mess. PMID- 12775638 TI - Doctors and managers: NHS needs political administration, not "management". PMID- 12775639 TI - Doctors and managers: Commission for Health Improvement gives its perspective. PMID- 12775640 TI - Doctors and managers: clear strategy is needed, as Turkish experience shows. PMID- 12775641 TI - Doctors and managers: healthcare management is treated as specialty in Israel. PMID- 12775642 TI - Discrimination in discretionary points award scheme: premise is flawed. PMID- 12775643 TI - Doctors and managers: what is the evidence base for management? PMID- 12775644 TI - Discrimination in discretionary points award scheme: discretionary points have to be applied for. PMID- 12775645 TI - Bioethics are difficult to balance. PMID- 12775646 TI - Walk-in centres are often first line contact for rape victims. PMID- 12775647 TI - GMC guidance on withholding life prolonging treatment. PMID- 12775648 TI - Cognitive behaviour therapy in practice. PMID- 12775649 TI - Clinical neurophysiology. PMID- 12775650 TI - Continuing professional development: choosing the right activity. PMID- 12775651 TI - Disabled and working. PMID- 12775652 TI - The impact of a journal and its editor on a career in experimental cardiology. PMID- 12775653 TI - Dissecting cardiac hypertrophy and signaling pathways: evidence for an interaction between multifunctional g proteins and prostanoids. PMID- 12775654 TI - Fibrillation or neurillation: back to the future in our concepts of sudden cardiac death? PMID- 12775655 TI - Tissue engineering therapy for cardiovascular disease. AB - The present treatments for the loss or failure of cardiovascular function include organ transplantation, surgical reconstruction, mechanical or synthetic devices, or the administration of metabolic products. Although routinely used, these treatments are not without constraints and complications. The emerging and interdisciplinary field of tissue engineering has evolved to provide solutions to tissue creation and repair. Tissue engineering applies the principles of engineering, material science, and biology toward the development of biological substitutes that restore, maintain, or improve tissue function. Progress has been made in engineering the various components of the cardiovascular system, including blood vessels, heart valves, and cardiac muscle. Many pivotal studies have been performed in recent years that may support the move toward the widespread application of tissue-engineered therapy for cardiovascular diseases. The studies discussed include endothelial cell seeding of vascular grafts, tissue engineered vascular conduits, generation of heart valve leaflets, cardiomyoplasty, genetic manipulation, and in vitro conditions for optimizing tissue-engineered cardiovascular constructs. PMID- 12775656 TI - Roles of cardiac transcription factors in cardiac hypertrophy. AB - Different cell types, equipped with unique structure and function, synthesize different sets of proteins on the basis of different patterns of gene expression, even though their genomes are identical. Cardiac transcription factors have been reported to control a cardiac gene program and thus to play a crucial role in transcriptional regulation during embryogenesis. Recently, postnatal roles of cardiac transcription factors have been extensively investigated. Consistent with the direct transactivation of numerous cardiac genes reactivated in response to hypertrophic stimulation, cardiac transcription factors are profoundly involved in the generation of cardiac hypertrophy or in cardioprotection from cytotoxic stress in the adult heart. In this review, the regulation of a cardiac gene program by cardiac transcription factors is summarized, with an emphasis on their potential role in the generation of cardiac hypertrophy. PMID- 12775659 TI - Radiological features of glandular odontogenic cyst. AB - OBJECTIVES: To present five new cases of glandular odontogenic cyst (GOC) and to review the radiological features at presentation as reported in the English literature. METHODS: From 1993 to 2002, five patients in our department were diagnosed with GOC based on histopathological findings and supported by radiography and CT. The radiographic features of the new GOC cases were analysed in addition to 51 literature cases. RESULTS: There were 31 male and 25 female patients, aged 14-90 years (mean 50 years). The mandible was involved in 80% and the maxilla in 20%; most of the lesions were located in the anterior jaw. Radiographically, 52% of the lesions were unilocular and 48% were multilocular; 94.5% showed well defined borders, which were sclerotic in 7.7% and scalloped in 13%. Information on cortical plate integrity was available in only 24 cases: 50% showed perforation, 8.3% erosion of the cortical plates and 8.3% thinning of the cortical plates. Root resorption was reported in 22% of patients and tooth displacement in 24.4%. CONCLUSIONS: Data collected indicate that GOC has potentially aggressive behaviour, with expansion and perforation in a significant number of cases. We recommend the use of multiple plane radiographs, with CT reserved for large lesions, especially those that are multilocular or involve extragnathic structures. PMID- 12775660 TI - Accuracy of panoramic radiography in assessing the dimensions of radiolucent jaw lesions with distinct or indistinct borders. AB - OBJECTIVES: To evaluate the accuracy of panoramic radiography (PR) in assessing the dimensions of mandibular lesions. METHODS: One hundred and fifteen cases exhibiting radiolucent lesions in the mandibular premolar, molar or ramus area were selected retrospectively. They were divided into four types: cyst with sclerotic margin (Type I); cyst without sclerotic margin (Type II); ameloblastoma (Type III); or squamous cell carcinoma (Type IV). Maximum mesiodistal length and superoinferior height were measured on PR (Lpmax and Hpmax, respectively) and on CT (Lcmax and Hcmax, respectively) and the results were compared. RESULTS: Correlation coefficients between Lpmax and Lcmax and between Hpmax and Hcmax were high for Type I, II and III lesions but were significantly lower for Type IV lesions. Regression coefficients between Lpmax and Lcmax and between Hpmax and Hcmax were indistinguishable from 1.0 for all types of lesions. The mean relative difference between Lpmax and Lcmax varied from 1.2% to 8.2%. The difference was only -0.3% for larger lesions of combined Types I and II. The mean relative difference between Hpmax and Hcmax varied from--3.5% to 1.1% depending on the type of lesions. CONCLUSION: PR is accurate for assessing the dimensions of radiolucent lesions in the posterior mandible when the margins are well defined. PMID- 12775661 TI - Curvature analysis of the mandibular condyle. AB - OBJECTIVE: To determine whether curvature analysis on high resolution CT images can be used as a tool for evaluation of mandibular condyle morphology. METHODS: Curvature analysis was performed on reconstructed oblique coronal CT images of 634 normal condyles from 317 patients (144 men and 173 women; age range 4-89 years) with inner or middle ear disease. The condyles were scanned with 1 mm collimation using helical CT. The CT images were analysed manually on a personal computer. RESULTS: The condyle CT images could be categorized into five curvature profile patterns: (1) bi-peak; (2) no peak; (3) tri-peak; (4) bi-peak with an intervening bottom above the base line (bi-peak with a col); and (5) bi-peak with an intervening negative phase. A separate evaluation using computer graphic condyle models indicated that these curvature patterns corresponded to flat (bi peak), round (no peak), convex (bi-peak with a col), concave (bi-peak with a negative phase) and angled (tri-peak) contours of the condyle's superior surface. The curvature profiles were identical between bilateral condyles in 40% (126/317) of the patients. Gender-related differences in the incidence of the curvature profiles were also found, the bi-peak with a col profile being more frequently observed in women and the bi-peak with a negative phase profile being observed more frequently in men. CONCLUSION: Curvature analysis on CT images depicts condyle morphology effectively and may be an adjunctive tool for condyle morphometry. PMID- 12775662 TI - A method for integrating facial cephalometry and corrected lateral tomography of the temporomandibular joint. AB - OBJECTIVES: To test the validity of an indirect method for obtaining the Frankfort horizontal (FH) plane and transferring it from the originating cephalogram to a tomogram. METHODS: One hundred and fifty-two corrected lateral tomograms from 38 individuals, taken before and after treatment, were used for this study. The tomograms were scanned and digitized. The angle between the FH plane and the long axis of the pterygopalatine fossa was measured on the reciprocal cephalograms. In addition, a tangent was drawn at the uppermost point of the glenoid fossa and parallel to the upper border of the film. Common variables regarding these two planes were measured and compared. RESULTS: It was found that the tangent to the fossa roof, parallel to the superior border of the film, did not coincide with the transferred cephalometric FH plane. Statistically significant differences were found between all variables for the two groups. CONCLUSIONS: In cases where integration of cephalometric and tomographic measurements are needed, the drawing of a tangent on the roof of the fossa is not a reliable method. PMID- 12775663 TI - Radiographic assessment of the marginal bone level after implant treatment: a comparison of periapical and Scanora detailed narrow beam radiography. AB - OBJECTIVES: To compare assessments of the marginal bone level around dental implants in the mandible using periapical radiography and Scanora detailed narrow beam (DNB) radiography. METHODS: Forty patients treated with Brnemark dental implants in the lower jaw were examined with periapical and Scanora DNB radiography. Ten implants were selected from each of the four dental regions (molar, incisor, canine, premolar), and no more than one implant was selected from the same patient. Seven observers assessed the level of the marginal bone on the mesial and distal surfaces of the implants. Three of the observers made all the assessments twice. RESULTS: Agreement between the methods was 61%. The highest agreement was found in the molar region. In DNB radiography the marginal bone level was observed to be situated more "coronally" in 17% and more "apically" in 22% compared with periapical radiography. The kappa value for interobserver agreement for all observers was 0.33 for periapical radiography and 0.27 for DNB radiography. The weighted kappa value for intraobserver agreement ranged from 0.75 to 0.99 for DNB radiography and from 0.94 to 0.98 for periapical radiography. CONCLUSIONS: Scanora multimodal radiography simplifies examination of implants in the mandible, and observer variation is comparable with that in intraoral periapical radiography. PMID- 12775664 TI - Radiographic alveolar bone loss in untreated Taiwan Chinese subjects with adult periodontitis measured by the digital scanning radiographic image analysis method. AB - OBJECTIVES: To determine differences in radiographic alveolar bone loss (RABL) by age group, gender and tooth type in subjects with adult periodontitis using the digital scanning radiographic image analysis (DSRIA) method. METHODS: A total of 4262 teeth were examined for RABL in 178 individuals (96 males and 82 females). Mesial and distal sites were determined by the DSRIA method following scanning of periapical radiographs and using the Microstation 95 image software. RESULTS: Linear assessment of RABL using DSRIA revealed the following results. Mean bone loss of maxillary and mandibular anterior teeth was significantly higher (P<0.001 and P<0.01, respectively) in males compared with females using a two-sample t test. The highest values of mean RABL occurred most commonly in the maxillary first molars (53.8% and 51.4% for females and males, respectively) and the mandibular central incisors (50.4% and 41.5% for males and females, respectively). Mean RABL was greater in males than in females for both the mandibular and maxillary arches, with the exception of molar sites. CONCLUSIONS: Maxillary first molars and mandibular incisors are the highest risk sites. Moreover, greater loss occurs at the molar site in females compared with males for untreated periodontitis in Taiwan Chinese people. PMID- 12775665 TI - Accuracy of digital and film panoramic radiographs for assessment of position and morphology of mandibular third molars and prevalence of dental anomalies and pathologies. AB - OBJECTIVES: To compare the accuracy of digital and film panoramic radiographs for determining (1) the position and morphology of mandibular third molars before surgical removal and (2) the prevalence of dental anomalies and pathologies. METHODS: Three hundred and eighty-eight third mandibular molars were available for examination. Position and morphology of third molars observed on film radiographs and on digital panoramic images from five systems (DenOptix, DigiDent, Digora, Dimax2 and Orthophos Plus) were recorded by two observers and were compared with surgeons' findings at the time of the operation (gold standard). One observer further recorded the prevalence of dental anomalies and pathologies on both imaging modalities. RESULTS: Few differences were found between the digital and film-based panoramic systems in the assessment of accuracy of position and morphology of mandibular third molars. The prevalence of dental anomalies and pathologies determined with the two modalities was roughly similar. CONCLUSION: The five digital panoramic systems evaluated in this study were equally as useful for third molar treatment planning and diagnosis of dental anomalies and pathologies as conventional film-based panoramic radiographs. PMID- 12775666 TI - Randomized controlled trial of a computer-assisted learning program to improve caries detection from bitewing radiographs. AB - OBJECTIVES: To investigate whether using a computer-assisted learning (CAL) calibration program improves the accuracy of dental students in caries detection from bitewing radiographs. METHODS: Dental students were assigned to an experimental (n=33) and control (n=34) group using a randomized block design. The experimental group used the CAL program with feedback to calibrate themselves against experts in radiographic caries detection and a histological gold standard of the actual clinical condition. Feedback was provided visually of the actual tooth surface condition and in the form of graphics showing diagnostic accuracy performance measures. Performance was tested before the program (for the control group) and after the program (for the experimental group) by assessing surfaces (n=56) from a new independent digital test set of evaluation radiographs for the presence, histologically, of dentine caries (n=23). The summary receiver operating characteristic (SROC) method for summarizing true positive ratio (TPR) (sensitivity) and false positive ratio (FPR) (1-specificity) was used to analyse the dichotomous data. Differences between the control and experimental groups were tested for (a) the area under the SROC curve (A(z)) and (b) the TPR, FPR and diagnostic odds ratio (DOR) using the Mann-Whitney test (P<0.05). RESULTS: The mean TPR for dentine caries detection was 76.3% (SD 13.0%) for the experimental group and 66.9% (SD 14.8%) for the control group (P=0.005). Mean FPRs were similar (experimental 28.1% and control 28.7%; P>0.5). The area under the SROC curve A(z) was 0.832 for the experimental group and 0.773 for the control group (P=0.002). The mean DOR for dentine caries in the experimental group (12.4) was better than that in the control group (8.8) (P=0.003). CONCLUSIONS: The CAL program does improve diagnostic performance. Improving the cognitive feedback provided by the program should be considered before implementation. PMID- 12775667 TI - Does digital radiography increase the number of intraoral radiographs? A questionnaire study of Dutch dental practices. AB - OBJECTIVES: To compare the number of radiographs taken in general dental practices equipped with digital radiography vs conventional film-based radiography and to determine the reasons for any difference in numbers. METHODS: In a mail survey, 473 questionnaires were sent to Dutch General Dental Practitioners (GDPs) using digital radiography and 105 questionnaires were sent to GDPs using film. The questionnaire concerned the number of intraoral radiographs taken in the dental practice, as well as possible reasons to take more or fewer radiographs after conversion to digital radiography. RESULTS: The response rate was 73%. Users of a phosphor plate system on average take 42.8 radiographs per week and solid-state system users take 48.4 radiographs, whereas film users take on average only 32.5 radiographs per week. The need for more certainty about the planned or ongoing treatment as well as better diagnostics were the most important reasons for taking more radiographs. CONCLUSIONS: It seems that GDPs using a system for digital radiography are more inclined to take radiographs than dentists taking conventional radiographs. Although digital intraoral radiography requires 50-80% less radiation per exposure than film, it is likely that the effective dose reduction after converting from conventional to digital radiography is less than 25% owing to the greater numbers of radiographs taken. PMID- 12775668 TI - Radiation doses of collimated vs non-collimated cephalometric exposures. AB - OBJECTIVES: The aim of the study was to compare organ and effective radiation doses for collimated and non-collimated cephalometric radiographs made by a multipurpose extraoral radiation unit. METHODS: Cephalometric exposures were made from a Rando head phantom representing an average man with the Cranex Tome multipurpose radiation unit. A collimator was fabricated from a 3-mm thick piece of lead. The collimator was designed so that structures relevant for orthodontic diagnosis were not shielded, whereas the thyroid gland and the major part of the skull were shielded. Absorbed organ doses were measured using thermoluminescent dosimeters. The effective dose was calculated both with (E(sal)) and without (E) inclusion of the salivary glands. RESULTS: Use of a wedge-shaped lead collimator yielded a reduction in field size of 55% and a reduction in E from 3.0 micro Sv to 1.6 micro Sv (47%) and in E(sal) from 3.7 micro Sv to 2.2 micro Sv (41%). CONCLUSIONS: Use of a wedge-shaped collimator should be advocated in all cases where depiction of the thyroid gland and the neurocranium is not required. The resulting reduction in effective dose using our collimator was 47% (41% when the salivary glands are included), which is of clinical importance, especially in children. PMID- 12775669 TI - Adenocarcinoma of the parotid gland with calcification. AB - A 78-year-old Korean woman was referred to Chonbuk National University Dental Hospital complaining of facial palsy and palpable mass in the right parotid gland area for 4 years. Clinical examination showed an asymmetrical facial appearance due to a 4 cmx5 cm hard, fixed, non-tender mass in the right parotid gland area, incomplete eye closure and a slight tremor at the corner of the mouth. A panoramic radiograph showed an amorphous calcified mass on the posterior mandibular ramus with thinning of the cortical plate adjacent to the mass. A sialogram showed constriction of the main duct and no further filling of striated, intercalated ducts and parenchymal areas. CT indicated an expansile mass with slight contrast enhancement involving the right parotid gland. The large mass showed necrotic areas and calcifications. A bone scan showed marked accumulation of (99)Tc(m)-methylene diphosphonate on the right posterior maxilla. Microscopic findings revealed minimal morphological alterations and rare mitotic figures within tumour cells, and the lesion was diagnosed as adenocarcinoma not otherwise specified (NOS, grade II). PMID- 12775670 TI - Susceptibility of Mycobacterium tuberculosis to weak acids. AB - The susceptibility of Mycobacterium tuberculosis and Mycobacterium smegmatis to a range of weak acids and acid pH was investigated. M. tuberculosis was found to be more susceptible to acid pH and weak acids than M. smegmatis. The weak acids were more active against M. tuberculosis at acid pH than at neutral pH. M. tuberculosis was found to be less able to maintain its internal pH and membrane potential at acid pH than M. smegmatis. The antituberculous activity of weak acids correlated with their ability to disrupt the membrane potential but not the internal pH. The significance of these findings is discussed in relation to M. tuberculosis physiology and development of new antituberculous agents. PMID- 12775671 TI - A review of the antimicrobial activity of clavulanate. AB - Clavulanate is a broad-spectrum beta-lactamase inhibitor, with activity against many of the chromosomally and plasmid-mediated beta-lactamases of both Gram positive and Gram-negative bacteria. Although clavulanate has minimal antibacterial activity in vitro, accumulating evidence suggests that it may have an effect on pathogenic bacteria regardless of beta-lactamase production. Like other beta-lactams, clavulanate has been shown to bind to penicillin-binding proteins (PBPs) in Gram-positive and Gram-negative bacteria. It was found to bind selectively to PBP3 in Streptococcus pneumoniae. It has been suggested that complementary binding to different PBPs and subsequent effects on autolysis contribute to the enhancement of the activity of other beta-lactams by clavulanate. In addition, co-amoxiclav has been shown to enhance the intracellular killing functions of human polymorphonuclear cells (PMNs) in studies undertaken with beta-lactamase-producing and non-beta-lactamase-producing strains of bacteria. These data from in vitro and cell culture systems have been reflected in vivo, where clavulanate enhanced the activity of amoxicillin against non-beta-lactamase-producing organisms. Further studies are required to determine whether the effects seen within in vitro and in vivo animal studies have clinical significance. PMID- 12775672 TI - Evaluation of the post-antifungal effect (PAFE) of amphotericin B and nystatin against 30 zygomycetes using two different media. AB - The post-antifungal effect (PAFE) of amphotericin B and nystatin against 30 clinical zygomycetes was evaluated using two different media. PAFE is a suppression of fungal growth after limited drug exposure. The MICs of both drugs were determined using NCCLS M38-P guidelines. A spectrophotometric method was used to determine PAFE in vitro. Spores were exposed to amphotericin B and nystatin in RPMI-1640 or AM3 at concentrations of 4 x and 1 x MIC for 4 h for Absidia sp. and at 1 x and 0.5 x MIC for 1 h for the other strains. Drugs were eliminated by washing. Exposed and control spores were cultured in microtitre wells and incubated for 48 h. PAFE was calculated as T - C (Delta t) between the control and the exposure fungi. The first increase in optical density (OD0) was used to calculate PAFE and was considered significant when the value of the lower 95%CI of the exposed strain was greater than the upper 95%CI of the control. MIC ranges in RPMI-1640 were: 0.06-4 mg/L for amphotericin B and 0.5-8 mg/L for nystatin; MIC ranges in AM3 were: 0.06-2 mg/L for amphotericin B and 0.5-4 mg/L for nystatin. Killing was not observed at the concentration and exposure time used. In RPMI-1640, for amphotericin B the rank order for PAFE was Absidia corymbifera (5.6 h) > Rhizopus oryzae (5.2 h) > Mucor spp. (3.5 h) > Rhizopus microsporus (3 h), and for nystatin the rank order was Mucor spp. (5.8 h) > R. oryzae (3.3 h) > A. corymbifera (2.9 h) > R. microsporus (1.7 h). PAFE was not induced in Rhizomucor spp. PAFE was dependent on drug concentration. PMID- 12775673 TI - AmpC beta-lactamases: what do we need to know for the future? PMID- 12775674 TI - Antiviral function of pyrrolidine dithiocarbamate against influenza virus: the inhibition of viral gene replication and transcription. PMID- 12775675 TI - Multiple antibiotic-resistance mechanisms including a novel combination of extended-spectrum beta-lactamases in a Klebsiella pneumoniae clinical strain isolated in Argentina. AB - Klebsiella pneumoniae M1803, isolated from a paediatric patient with chronic urinary infection, presented nine antimicrobial resistance mechanisms harboured on two conjugative megaplasmids, in addition to the chromosomally mediated SHV-1 beta-lactamase. These nine antimicrobial resistance mechanisms comprised two extended-spectrum beta-lactamases (ESBLs) (PER-2 and CTX-M-2), TEM-1-like, OXA-9 like, AAC(3)-IIa, AAC(6')-Ib, ANT(3")-Ia and resistance determinants to tetracycline and chloramphenicol. During fluoroquinolone treatment, a variant derived from M1803 (named M1826) was selected, with an overall increase of MICs, in particular of cefoxitin and carbapenems. No enzymic activity against these latter drugs was found. Mutations in the region analogous to the quinolone resistance-determining region were not found. Strain M1826 was deficient in OmpK35/36 expression, which produced the decrease in the susceptibility to cefoxitin, carbapenems and fluoroquinolones. The blaCTX-M-2 gene was located in an unusual class 1 integron, which includes Orf513, as occurred in the recently described In35. In addition, Tn3 and Tn1331 were detected in both K. pneumoniae isolates. This is the first report of in vivo selection of an OmpK35/36 deficiency in a K. pneumoniae strain that produced a novel combination of two ESBLs (CTX-M-2 and PER-2) during fluoroquinolone treatment in a paediatric patient with chronic urinary infection. PMID- 12775676 TI - Determination by HPLC of chlortetracycline in pig faeces. AB - An HPLC assay used to determine chlortetracycline (CTC) in pig faeces is reported. Prodigy ODS3 (4.6 x 150 mm) was used for the stationary phase, whereas the mobile phase comprised oxalic acid, sodium oxalate and sodium decane sulfonate (66%)--each of 4 mM, and 34% acetonitrile. The mobile phase was pumped at a flow rate of 1 mL/min. Detection of CTC was by ultraviolet absorbance at 370 nm, and a 20 micro L injection volume was used. Recovery from faeces was >90%, and coefficients of variability between runs were <10%. The lowest limit of quantification was 3.5 mg/kg, with an accuracy of <7% error. There was no interference from endogenous materials in the pig faeces, or commonly used antibiotics, and the method is suitable for use in drug disposition studies. PMID- 12775677 TI - Pharmacodynamic activity of azithromycin against macrolide-susceptible and resistant Streptococcus pneumoniae simulating clinically achievable free serum, epithelial lining fluid and middle ear fluid concentrations. AB - BACKGROUND: The association between macrolide resistance mechanisms and bacteriological eradication of Streptococcus pneumoniae remains poorly studied. The present study, using an in vitro pharmacodynamic model, assessed azithromycin activity against macrolide-susceptible and -resistant S. pneumoniae simulating clinically achievable free serum (S), epithelial lining fluid (ELF) and middle ear fluid (MEF) concentrations. MATERIALS AND METHODS: Two macrolide-susceptible [PCR-negative for both mef(A) and erm(B)] and six macrolide-resistant [five mef(A)-positive/erm(B)-negative displaying various degrees of macrolide resistance and one mef(A)-negative/erm(B)-positive] S. pneumoniae were tested. Azithromycin was modelled simulating a dosage of 500 mg/250 mg by mouth, once a day [free S: maximum concentration (Cmax) 0.2 mg/L, t1/2 68 h; free ELF Cmax 1.0 mg/L, t1/2 68 h] and 10 mg/kg by mouth, once a day (free MEF: Cmax 1.0 mg/L, t1/2 68 h) using a one compartment model. Starting inocula were 1 x 10(6) cfu/mL in Mueller-Hinton broth with 2% lysed horse blood. Sampling at 0, 2, 4, 6, 12, 24 and 48 h assessed the extent of bacterial killing (decrease in log10 cfu/mL versus initial inoculum). RESULTS: Free azithromycin concentrations in serum, ELF and MEF simulating time above the MIC (T > MIC) of 100% [area under the curve to MIC (AUC0-24/MIC] > or = 36.7] were bactericidal (> or = 3 log10 killing) at 24 and 48 h versus macrolide-susceptible S. pneumoniae. Against macrolide-resistant S. pneumoniae, free serum concentrations providing T > MIC of 0% or AUC0-24/MIC < or = 1.1 demonstrated no bacterial inhibition followed by regrowth at 24 and 48 h, whereas free ELF and MEF providing T > MIC of 0% or AUC0-24/MIC of 4.6 produced a bacteriostatic (0.2-0.5 log10 killing at 24 h) effect with a mef(A) strain with an azithromycin MIC of 2 mg/L. Against mef(A)-positive S. pneumoniae strains with azithromycin MICs > or = 4 mg/L, no bacterial killing occurred at any time point and rapid regrowth was observed simulating ELF or MEF T > MIC of 0% or AUC0-24/MIC < or = 2.3. CONCLUSION: Azithromycin serum, ELF and MEF concentrations rapidly eradicated macrolide-susceptible S. pneumoniae but did not eradicate macrolide-resistant S. pneumoniae regardless of resistance phenotype. PMID- 12775678 TI - Comparative efficacy of daptomycin and vancomycin in the therapy of experimental foreign body infection due to Staphylococcus aureus. AB - The therapeutic activity of daptomycin was compared with that of vancomycin in a rat model of subcutaneously implanted tissue cages chronically infected with strain Rev1, a spontaneous methicillin-susceptible revertant of the methicillin resistant Staphylococcus aureus strain MRGR3, showing equivalent virulence to its parent. The MIC and MBC of daptomycin (in Mueller-Hinton broth supplemented with 50 mg/L Ca2+) or vancomycin for strain Rev1 were 1-2 and 2-4 or 1 and 2 mg/L, respectively. In vitro elimination of strain Rev1 in the presence of 50% tissue cage fluid was more rapid with daptomycin 4 mg/L compared with vancomycin. After 2 weeks of infection, viable counts of strain Rev1 averaged 6.49 log10 cfu/mL of tissue cage fluid (n = 87). Intraperitoneal administration of daptomycin 30 mg/kg once daily, or vancomycin 50 mg/kg twice daily, produced antibiotic levels continuously above MBC. After 7 days of therapy with daptomycin or vancomycin, mean +/- S.E.M. counts of Rev1 decreased (P < 0.05) by 1.11 +/- 0.25 (n = 28) or 0.80 +/- 0.31 (n = 35) log10 cfu/mL, respectively, compared with cages of untreated animals, but were not significantly different from each other. In daptomycin-treated rats, three cages yielded subpopulations with reduced susceptibility to daptomycin. In conclusion, a low dose regimen of daptomycin was at least equivalent to vancomycin against chronic foreign body infections due to S. aureus. Drug dosage should be adapted to obtain inflammatory fluid levels of daptomycin minimizing emergence of resistant subpopulations. PMID- 12775679 TI - Endemic carbapenem-resistant Acinetobacter baumannii in a London hospital. PMID- 12775680 TI - Stable susceptibility of Candida blood isolates to fluconazole despite increasing use during the past 10 years. AB - The prevalence of drug-resistant bacterial pathogens is very high in Taiwan. Accordingly, there was great concern that the introduction of fluconazole would result in rapid emergence of drug-resistant yeasts. Thus, we recommended in 1991 that fluconazole be used for treatment only. To explore the impact of this policy fluconazole susceptibility of Candida species blood culture isolates and outcome of patients with nosocomial candidaemia were monitored prospectively at National Taiwan University Hospital during 1994-2000. The MICs of fluconazole were determined by the disc diffusion method. There were 1095 episodes of nosocomial candidaemia during 1994-2000. Candida albicans was the most common species (50.4%), followed by Candida tropicalis (20.5%), Candida parapsilosis (14.2%) and Candida glabrata (12.0%). There were 0-2 isolates of Candida krusei per year. The incidence of nosocomial candidaemia and the proportion of C. glabrata peaked in 1996 and decreased thereafter. Fluconazole susceptibility was determined for 552 Candida blood isolates. Only 0.7% of blood isolates were resistant to fluconazole. Fluconazole susceptibility was 94.0% in 1994-1995 and 97.9% in 1999 2000 (P = 0.06). Attributable mortality for patients with nosocomial candidaemia was 43.2% in 1994-1995 and was 25% in 2000 (P = 0.005). Despite an increase in the incidence of nosocomial fungal infection and increased consumption of fluconazole from 1994 to 2000, there was no significant change in the susceptibility to fluconazole for bloodstream isolates of Candida species. These findings appear to be attributed to several factors. These include low prevalence of C. krusei and C. glabrata, changing patterns of use of antifungal drugs and broad-spectrum antibiotics, and efforts to improve the rational use of antifungal agents at our hospital. PMID- 12775681 TI - Increasing prevalence of antimicrobial resistance among Pseudomonas aeruginosa isolates in Latin American medical centres: 5 year report of the SENTRY Antimicrobial Surveillance Program (1997-2001). PMID- 12775682 TI - Effects of cefotaxime and desacetylcefotaxime upon Clostridium difficile proliferation and toxin production in a triple-stage chemostat model of the human gut. AB - Clostridium difficile is recognized as an important nosocomial pathogen. C. difficile infection (CDI) is thought to arise as a result of depletion of the normal gut flora by antimicrobial agents. Cefotaxime (CTX) is well-known for its propensity to cause CDI, but the reasons behind its particular predisposition to the disease remain unclear. Previous investigations have so far relied upon the hamster model of CDI or human volunteers. We have used a triple-stage chemostat model of the human gut to investigate the behaviour of C. difficile and components of the normal gut flora, in response to exposure to CTX alone, and in combination with its active metabolite desacetylcefotaxime (dCTX). C. difficile remained in a steady state during non-antibiotic exposed periods, with no detectable cytotoxin. During both antibiotic exposure regimens, proliferation of C. difficile and elevated cytotoxin levels were observed. Cessation of antibiotic instillation produced a reduction in cytotoxin levels and viable counts. Decreases in bacterial counts were observed in response to both antibiotic exposure regimens, notably for bifidobacteria and bacteroides. Numbers of bacteroides were profoundly affected by exposure to the CTX/dCTX combination, and this may indicate a possible role for bacteroides in colonization resistance. We believe that the gut model is a promising method for studying C. difficile pathogenesis in conditions analogous to the in vivo situation. PMID- 12775683 TI - Effect of D240G substitution in a novel ESBL CTX-M-27. AB - Escherichia coli clinical strain Gre-1 collected in 2000 from a French hospital harboured a novel CTX-M-encoding gene, designated blaCTX-M-27. CTX-M-27 differed from CTX-M-14 only by the substitution D240G and was the third CTX-M enzyme harbouring this mutation after CTX-M-15 and CTX-M-16. The Gly-240-harbouring enzyme CTX-M-27 conferred to E. coli higher MICs of ceftazidime (MIC, 8 versus 1 mg/L) than did the Asp-240-harbouring CTX-M-14 enzyme. Comparison of CTX-M-14 and CTX-M-27 showed that residue Gly-240 decreased Km for ceftazidime (205 versus 940 microM), but decreased hydrolytic activity against good substrates, such as cefotaxime (kcat, 113 versus 415 s-1), probably owing to the alteration of beta3 strand positioning during the catalytic process. PMID- 12775684 TI - Purification and properties of an intracellular 3-hydroxybutyrate-oligomer hydrolase (PhaZ2) in Ralstonia eutropha H16 and its identification as a novel intracellular poly(3-hydroxybutyrate) depolymerase. AB - An intracellular 3-hydroxybutyrate (3HB)-oligomer hydrolase (PhaZ2(Reu)) of Ralstonia eutropha was purified from Escherichia coli harboring a plasmid containing phaZ2(Reu). The purified enzyme hydrolyzed linear and cyclic 3HB oligomers. Although it did not degrade crystalline poly(3-hydroxybutyrate) (PHB), the purified enzyme degraded artificial amorphous PHB at a rate similar to that of the previously identified intracellular PHB (iPHB) depolymerase (PhaZ1(Reu)). The enzyme appeared to be an endo-type hydrolase, since it actively hydrolyzed cyclic 3HB-oligomers. However, it degraded various linear 3HB-oligomers and amorphous PHB in the fashion of an exo-type hydrolase, releasing one monomer unit at a time. PhaZ2 was found to bind to PHB inclusion bodies and as a soluble enzyme to cell-free supernatant fractions in R. eutropha; in contrast, PhaZ1 bound exclusively to the inclusion bodies. When R. eutropha H16 was cultivated in a nutrient-rich medium, the transient deposition of PHB was observed: the content of PHB was maximized in the log growth phase (12 h, ca. 14% PHB of dry cell weight) and decreased to a very low level in the stationary phase (ca. 1% of dry cell weight). In each phaZ1-null mutant and phaZ2-null mutant, the PHB content in the cell increased to ca. 5% in the stationary phase. A double mutant lacking both phaZ1 and phaZ2 showed increased PHB content in the log phase (ca. 20%) and also an elevated PHB level (ca. 8%) in the stationary phase. These results indicate that PhaZ2 is a novel iPHB depolymerase, which participates in the mobilization of PHB in R. eutropha along with PhaZ1. PMID- 12775685 TI - SigM, an extracytoplasmic function sigma factor of Bacillus subtilis, is activated in response to cell wall antibiotics, ethanol, heat, acid, and superoxide stress. AB - The extracytoplasmic function sigma M of Bacillus subtilis is required for normal cell growth under salt stress. It is expressed maximally during exponential growth and is further induced by the addition of 0.7 M NaCl. The promoter region of the sigM operon contains two promoters; one (P(A)) is sigma A dependent, and the other (P(M)) is sigma M dependent. These have been placed separately at the amy locus, directing expression of a lacZ reporter gene. Only the P(M) fusion responded to salt induction. This promoter, which was responsive to the level of active sigma M in the cell, was also induced by 5% ethanol, by vancomycin, bacitracin, or phosphomycin (inhibitors of cell wall biosynthesis; 2 micro g per ml), and by heat shock of 50 degrees C for 10 min. It was very strongly induced by acid (pH 4.3) and 80 micro M paraquat, but after a 15- to 30-min delay. There was no induction by alkali (pH 9), 5 mM H(2)O(2), the detergents 0.1% Triton X 100 and 0.1% Tween 20, or 50 micro M monensin. In addition to their reduced tolerance to salt, null mutants of sigM were unable to grow at pH 4.3 and lysed after exposure to 5% ethanol. Genes regulated by SigM were also tested for their response to pH 4.3, 5% ethanol, and 2 micro g of vancomycin per ml. Expression of the genes may have been activated by increased levels of sigma M, but at least some were also subject to additional controls, as they responded to one type of stress but not another. Expression of yrhJ, which encodes a cytochrome P450/NADPH reductase, was induced in response to acid and vancomycin. yraA expression was acid, ethanol, and vancomycin induced, whereas yjbD showed only ethanol induction. YraA protein was extremely important to acid survival-a mutation in yraA, like a sigM mutation, resulted in the failure of B. subtilis to grow at pH 4.3. Sigma M is therefore involved in maintaining membrane and cell wall integrity in response to several different stresses in exponential growth phase and is activated by such stresses. PMID- 12775686 TI - The ttsA gene is required for low-calcium-induced type III secretion of Yop proteins and virulence of Yersinia enterocolitica W22703. AB - Pathogenic Yersinia species use a virulence-plasmid encoded type III secretion pathway to escape the innate immune response and to establish infections in lymphoid tissues. At least 22 secretion machinery components are required for type III transport of 14 different Yop proteins, and 10 regulatory factors are responsible for activating this pathway in response to environmental signals. Although the genes for these products are located on the 70-kb virulence plasmid of Yersinia, this extrachromosomal element does not appear to harbor genes that provide for the sensing of environmental signals, such as calcium-, glutamate-, or serum-sensing proteins. To identify such genes, we screened transposon insertion mutants of Y. enterocolitica W22703 for defects in type III secretion and identified ttsA, a chromosomal gene encoding a polytopic membrane protein. ttsA mutant yersiniae synthesize reduced amounts of Yops and display a defect in low-calcium-induced type III secretion of Yop proteins. ttsA mutants are also severely impaired in bacterial motility, a phenotype which is likely due to the reduced expression of flagellar genes. All of these defects were restored by complementation with plasmid-encoded wild-type ttsA. LcrG is a repressor of the Yersinia type III pathway that is activated by an environmental calcium signal. Mutation of the lcrG gene in a ttsA mutant strain restored the type III secretion of Yop proteins, although the double mutant strain secreted Yops in the presence and absence of calcium, similar to the case for mutants that are defective in lcrG gene function alone. To examine the role of ttsA in the establishment of infection, we measured the bacterial dose required to produce an acute lethal disease following intraperitoneal infection of mice. The ttsA insertion caused a greater-than-3-log-unit reduction in virulence compared to that of the parental strain. PMID- 12775687 TI - PhoP-responsive expression of the Salmonella enterica serovar typhimurium slyA gene. AB - The SlyA protein of Salmonella enterica serovar Typhimurium is a member of the MarR family of transcription regulators and is required for virulence and survival in professional macrophages. Isolated SlyA protein was able to bind a specific DNA target without posttranslational modification. This suggested that SlyA might not be activated by directly sensing an external signal but rather that the intracellular concentration of SlyA is enhanced in appropriate environments through the action of other transcription factors. Analysis of slyA transcription reveals the presence of a promoter region located upstream of the previously recognized SlyA repressed promoter. The newly identified upstream promoter region did not respond to SlyA but was activated by Mg(II) starvation in a PhoP-dependent manner. We present here evidence for a direct link between two transcription factors (PhoP and SlyA) crucial for Salmonella virulence. PMID- 12775688 TI - The Pseudomonas fluorescens AlgG protein, but not its mannuronan C-5-epimerase activity, is needed for alginate polymer formation. AB - Bacterial alginates are produced as 1-4-linked beta-D-mannuronan, followed by epimerization of some of the mannuronic acid residues to alpha-L-guluronic acid. Here we report the isolation of four different epimerization-defective point mutants of the periplasmic Pseudomonas fluorescens mannuronan C-5-epimerase AlgG. All mutations affected amino acids conserved among AlgG-epimerases and were clustered in a part of the enzyme also sharing some sequence similarity to a group of secreted epimerases previously reported in Azotobacter vinelandii. An algG-deletion mutant was constructed and found to produce predominantly a dimer containing a 4-deoxy-L-erythro-hex-4-enepyranosyluronate residue at the nonreducing end and a mannuronic acid residue at the reducing end. The production of this dimer is the result of the activity of an alginate lyase, AlgL, whose in vivo activity is much more limited in the presence of AlgG. A strain expressing both an epimerase-defective (point mutation) and a wild-type epimerase was constructed and shown to produce two types of alginate molecules: one class being pure mannuronan and the other having the wild-type content of guluronic acid residues. This formation of two distinct classes of polymers in a genetically pure cell line can be explained by assuming that AlgG is part of a periplasmic protein complex. PMID- 12775689 TI - Protein synthesis in Escherichia coli with mischarged tRNA. AB - Two types of aspartyl-tRNA synthetase exist: the discriminating enzyme (D-AspRS) forms only Asp-tRNA(Asp), while the nondiscriminating one (ND-AspRS) also synthesizes Asp-tRNA(Asn), a required intermediate in protein synthesis in many organisms (but not in Escherichia coli). On the basis of the E. coli trpA34 missense mutant transformed with heterologous ND-aspS genes, we developed a system with which to measure the in vivo formation of Asp-tRNA(Asn) and its acceptance by elongation factor EF-Tu. While large amounts of Asp-tRNA(Asn) are detrimental to E. coli, smaller amounts support protein synthesis and allow the formation of up to 38% of the wild-type level of missense-suppressed tryptophan synthetase. PMID- 12775690 TI - The Stigmatella aurantiaca homolog of Myxococcus xanthus high-mobility-group A type transcription factor CarD: insights into the functional modules of CarD and their distribution in bacteria. AB - Transcriptional factor CarD is the only reported prokaryotic analog of eukaryotic high-mobility-group A (HMGA) proteins, in that it has contiguous acidic and AT hook DNA-binding segments and multifunctional roles in Myxococcus xanthus carotenogenesis and fruiting body formation. HMGA proteins are small, randomly structured, nonhistone, nuclear architectural factors that remodel DNA and chromatin structure. Here we report on a second AT hook protein, CarD(Sa), that is very similar to CarD and that occurs in the bacterium Stigmatella aurantiaca. CarD(Sa) has a C-terminal HMGA-like domain with three AT hooks and a highly acidic adjacent region with one predicted casein kinase II (CKII) phosphorylation site, compared to the four AT hooks and five CKII sites in CarD. Both proteins have a nearly identical 180-residue N-terminal segment that is absent in HMGA proteins. In vitro, CarD(Sa) exhibits the specific minor-groove binding to appropriately spaced AT-rich DNA that is characteristic of CarD or HMGA proteins, and it is also phosphorylated by CKII. In vivo, CarD(Sa) or a variant without the single CKII phosphorylation site can replace CarD in M. xanthus carotenogenesis and fruiting body formation. These two cellular processes absolutely require that the highly conserved N-terminal domain be present. Thus, three AT hooks are sufficient, the N-terminal domain is essential, and phosphorylation in the acidic region by a CKII-type kinase can be dispensed with for CarD function in M. xanthus carotenogenesis and fruiting body development. Whereas a number of hypothetical proteins homologous to the N-terminal region occur in a diverse array of bacterial species, eukaryotic HMGA-type domains appear to be confined primarily to myxobacteria. PMID- 12775691 TI - Relaxed specificity of the R1162 nickase: a model for evolution of a system for conjugative mobilization of plasmids. AB - The primary DNA processing protein for conjugative mobilization of the plasmid R1162 is the transesterase MobA, which acts at a unique site on the plasmid, the origin of transfer (oriT). Both MobA and oriT are members of a large family of related elements that are widely distributed among bacteria. Each oriT consists of a highly conserved core and an adjacent region that is required for binding by its cognate MobA. The sequence of the adjacent region is important in determining the specificity of the interaction between the Mob protein and the oriT DNA. However, the R1162 MobA is active on the oriT of pSC101, another naturally occurring plasmid. We show here that MobA can recognize oriTs having different sequences in the adjacent region and, with varying frequencies, can cleave these oriTs at the correct position within the core. Along with the structure of the oriTs themselves, these characteristics suggest a model for the evolution of this group of transfer systems. PMID- 12775692 TI - FlrA, a sigma54-dependent transcriptional activator in Vibrio fischeri, is required for motility and symbiotic light-organ colonization. AB - Flagellum-mediated motility of Vibrio fischeri is an essential factor in the bacterium's ability to colonize its host, the Hawaiian squid Euprymna scolopes. To begin characterizing the nature of the flagellar regulon, we have cloned a gene, designated flrA, from V. fischeri that encodes a putative sigma(54) dependent transcriptional activator. Genetic arrangement of the flrA locus in V. fischeri is similar to motility master-regulator operons of Vibrio cholerae and Vibrio parahaemolyticus. In addition, examination of regulatory regions of a number of flagellar operons in V. fischeri revealed apparent sigma(54) recognition motifs, suggesting that the flagellar regulatory hierarchy is controlled by a similar mechanism to that described in V. cholerae. However, in contrast to its closest known relatives, flrA mutant strains of V. fischeri ES114 were completely abolished in swimming capability. Although flrA provided in trans restored motility to the flrA mutant, the complemented strain was unable to reach wild-type levels of symbiotic colonization in juvenile squid, suggesting a possible role for the proper expression of FlrA in regulating symbiotic colonization factors in addition to those required for motility. Comparative RNA arbitrarily primed PCR analysis of the flrA mutant and its wild-type parent revealed several differentially expressed transcripts. These results define a regulon that includes both flagellar structural genes and other genes apparently not involved in flagellum elaboration or function. Thus, the transcriptional activator FlrA plays an essential role in regulating motility, and apparently in modulating other symbiotic functions, in V. fischeri. PMID- 12775693 TI - Posttranscriptional control of quorum-sensing-dependent virulence genes by DksA in Pseudomonas aeruginosa. AB - Pseudomonas aeruginosa controls the secretion of extracellular virulence factors, including rhamnolipids and LasB elastase, by the las and rhl quorum-sensing systems. Here, we mutated the dksA gene of P. aeruginosa by insertion of an Omega Hg cassette. The mutant displayed growth rates similar to that of the wild type in rich medium but was impaired in growth in defined minimal medium. Production of rhamnolipids and LasB elastase by the dksA mutant was only 4 and 10%, respectively, of wild-type levels. These defects could be partially complemented by introduction of the plasmid-encoded dksA genes from P. aeruginosa or Escherichia coli. In the dksA mutant, the expression of rhlI was increased early during exponential growth, but expression of other quorum-sensing regulator genes lasR, lasI, and rhlR-was not affected. Although the transcription of the lasB and rhlAB genes was comparable between the dksA mutant and the wild-type strain in peptone tryptic soy broth medium, we observed reduced translation of both genes in the dksA mutant. Similarly, we found that full translation of lasB and rhlAB genes in E. coli also requires the dksA gene. DksA is therefore a novel regulator involved in the posttranscriptional control of extracellular virulence factor production in P. aeruginosa. PMID- 12775694 TI - Flagellar phase variation in Salmonella enterica is mediated by a posttranscriptional control mechanism. AB - Salmonella enterica has two antigenically distinct flagellin genes, fliC and fljB, that are alternatively expressed. The fljA gene is cotranscribed with fljB and encodes a protein that has been characterized as a transcriptional repressor of the unlinked fliC gene when FljB is expressed. In this study we report genetic evidence that FljA prevents the production of FliC protein through an interaction with the 5'-untranslated region of the fliC mRNA transcript. Studies with operon and gene fusions, Western analyses, and T(2) RNase protection assays were performed for strains with the fljBA promoter locked in either the on or the off orientation. beta-Galactosidase assays of fliC transcriptional and translational fusions to the lac operon demonstrated that while FljA inhibits fliC transcription fivefold in the fljBA(ON) orientation, it has a 200-fold effect on both fliC transcription and translation, indicating that the FljA inhibitor might act at both the transcriptional and translational level. T(2) RNase protection assays also demonstrated a fivefold decrease in fliC transcript levels for cells locked in the fljBA(ON) orientation compared to those in the fljBA(OFF) orientation, and an eightfold decrease in FliC protein levels was observed by Western analysis. This reduction in FliC protein levels is greater than the decrease observed for the transcript. These results are consistent with a new model whereby FljA inhibits FliC expression by an attenuation or translational control mechanism. PMID- 12775695 TI - Characterization of the second LysR-type regulator in the biphenyl-catabolic gene cluster of Pseudomonas pseudoalcaligenes KF707. AB - Pseudomonas pseudoalcaligenes KF707 possesses a biphenyl-catabolic (bph) gene cluster consisting of bphR1A1A2-(orf3)-bphA3A4BCX0X1X2X3D. The bphR1 (formerly orf0) gene product, which belongs to the GntR family, is a positive regulator for itself and bphX0X1X2X3D. Further analysis in this study revealed that a second regulator belonging to the LysR family (designated bphR2) is involved in the regulation of the bph genes in KF707. The bphR2 gene was not located near the bph gene cluster, and its product (BphR2) exhibited a high level of similarity to NahR (the naphthalene- and salicylate-catabolic regulator belonging to the LysR family) in plasmid NAH7 of Pseudomonas putida. A strain containing a disrupted bphR2 gene failed to grow on biphenyl as a sole source of carbon, and the BphD (2 hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid hydrolase) activity was significantly reduced compared to that of wild-type strain KF707. Furthermore, the same strain exhibited extremely low transcription of bphR1, bphA1, bphC, bphX0, and bphD. However, when the bphR2 gene was provided in trans to the bphR2-disrupted strain, the transcription level of these genes was restored. These results indicate that bphR2 regulates the bph genes positively as a second regulator together with BphR1. PMID- 12775696 TI - Suppression of temperature-sensitive chromosome replication of an Escherichia coli dnaX(Ts) mutant by reduction of initiation efficiency. AB - Temperature sensitivity of DNA polymerization and growth of a dnaX(Ts) mutant is suppressible at 39 to 40 degrees C by mutations in the initiator gene, dnaA. These suppressor mutations concomitantly cause initiation inhibition at 20 degrees C and have been designated Cs,Sx to indicate both phenotypic characteristics of cold-sensitive initiation and suppression of dnaX(Ts). One dnaA(Cs,Sx) mutant, A213D, has reduced affinity for ATP, and two mutants, R432L and T435K, have eliminated detectable DnaA box binding in vitro. Two models have explained dnaA(Cs,Sx) suppression of dnaX, which codes for both the tau and gamma subunits of DNA polymerase III. The initiation deficiency model assumes that reducing initiation efficiency allows survival of the dnaX(Ts) mutant at the somewhat intermediate temperature of 39 to 40 degrees C by reducing chromosome content per cell, thus allowing partially active DNA polymerase III to complete replication of enough chromosomes for the organism to survive. The stabilization model is based on the idea that DnaA interacts, directly or indirectly, with polymerization factors during replication. We present five lines of evidence consistent with the initiation deficiency model. First, a dnaA(Cs,Sx) mutation reduced initiation frequency and chromosome content (measured by flow cytometry) and origin/terminus ratios (measured by real-time PCR) in both wild-type and dnaX(Ts) strains growing at 39 and 34 degrees C. These effects were shown to result specifically from the Cs,Sx mutations, because the dnaX(Ts) mutant is not defective in initiation. Second, reduction of the number of origins and chromosome content per cell was common to all three known suppressor mutations. Third, growing the dnaA(Cs,Sx) dnaX(Ts) strain on glycerol-containing medium reduced its chromosome content to one per cell and eliminated suppression at 39 degrees C, as would be expected if the combination of poor carbon source, the Cs,Sx mutation, the Ts mutation, and the 39 degrees C incubation reduced replication to the point that growth (and, therefore, suppression) was not possible. However, suppression was possible on glycerol medium at 38 degrees C. Fourth, the dnaX(Ts) mutation can be suppressed also by introduction of oriC mutations, which reduced initiation efficiency and chromosome number per cell, and the degree of suppression was proportional to the level of initiation defect. Fifth, introducing a dnaA(Cos) allele, which causes overinitiation, into the dnaX(Ts) mutant exacerbated its temperature sensitivity. PMID- 12775698 TI - Molecular characterization of inulosucrase from Leuconostoc citreum: a fructosyltransferase within a glucosyltransferase. AB - The gene coding for inulosucrase in Leuconostoc citreum CW28, islA, was cloned, sequenced, and expressed in Escherichia coli. The recombinant enzyme catalyzed inulin synthesis from sucrose like the wild-type enzyme. Inulosucrase presents an unusual structure: its N-terminal region is similar to the variable region of glucosyltransferases, its catalytic domain is similar to fructosyltransferases from various microorganisms, and its C-terminal domain presents similarity to the glucan binding domain from alternansucrase, a glucosyltransferase from Leuconostoc mesenteroides NRRL B-1355. From sequence comparison, it was found that this fructosyltransferase is a natural chimeric enzyme resulting from the substitution of the catalytic domain of alternansucrase by a fructosyltransferase. Two different forms of the islA gene truncated in the C terminal glucan binding domain were successfully expressed in E. coli and retained their ability to synthesize inulin but lost thermal stability. This is the first report of an inulosucrase bearing structural features of both glucosyltransferases and fructosyltransferases. PMID- 12775697 TI - Escherichia coli O157:H7 Shiga toxin-encoding bacteriophages: integrations, excisions, truncations, and evolutionary implications. AB - As it descended from Escherichia coli O55:H7, Shiga toxin (Stx)-producing E. coli (STEC) O157:H7 is believed to have acquired, in sequence, a bacteriophage encoding Stx2 and another encoding Stx1. Between these events, sorbitol fermenting E. coli O157:H(-) presumably diverged from this clade. We employed PCR and sequence analyses to investigate sites of bacteriophage integration into the chromosome, using evolutionarily informative STEC to trace the sequence of acquisition of elements encoding Stx. Contrary to expectations from the two currently sequenced strains, truncated bacteriophages occupy yehV in almost all E. coli O157:H7 strains that lack stx(1) (stx(1)-negative strains). Two truncated variants were determined to contain either GTT or TGACTGTT sequence, in lieu of 20,214 or 18,895 bp, respectively, of the bacteriophage central region. A single nucleotide polymorphism in the latter variant suggests that recombination in that element extended beyond the inserted octamer. An stx(2) bacteriophage usually occupies wrbA in stx(1)(+)/stx(2)(+) E. coli O157:H7, but wrbA is unexpectedly unoccupied in most stx(1)-negative/stx(2)(+) E. coli O157:H7 strains, the presumed progenitors of stx(1)(+)/stx(2)(+) E. coli O157:H7. Trimethoprim sulfamethoxazole promotes the excision of all, and ciprofloxacin and fosfomycin significantly promote the excision of a subset of complete and truncated stx bacteriophages from the E. coli O157:H7 strains tested; bile salts usually attenuate excision. These data demonstrate the unexpected diversity of the chromosomal architecture of E. coli O157:H7 (with novel truncated bacteriophages and multiple stx(2) bacteriophage insertion sites), suggest that stx(1) acquisition might be a multistep process, and compel the consideration of multiple exogenous factors, including antibiotics and bile, when chromosome stability is examined. PMID- 12775699 TI - Role of the Enterococcus faecalis GelE protease in determination of cellular chain length, supernatant pheromone levels, and degradation of fibrin and misfolded surface proteins. AB - Gelatinase (GelE), a secreted Zn-metalloprotease of Enterococcus faecalis, has been implicated as a virulence factor by both epidemiological data and animal model studies. Expression of gelE is induced at a high cell density by the fsr quorum-sensing system. In the present study, GelE was shown to be responsible for the instability of a number of Asc10 (aggregation substance) mutant proteins, implying that GelE functions to clear the bacterial cell surface of misfolded proteins. Disruption of GelE production led to increased cell chain length of E. faecalis, from a typical diplococcus morphology to chains of 5 to 10 cells. This function of GelE was also exhibited when the protein was expressed in Streptococcus pyogenes. GelE-expressing E. faecalis strains were more autolytic, suggesting that GelE affects chain length through activation of an autolysin. GelE was also essential for degradation of polymerized fibrin. GelE expression reduced the titer of cCF10, the peptide pheromone that induces conjugation of pCF10, and pCF10 had increased conjugation into non-GelE-expressing strains. These new functions attributed to GelE suggest that it acts to increase the dissemination of E. faecalis in high-density environments. PMID- 12775700 TI - Variation between pathogenic serovars within Salmonella pathogenicity islands. AB - Although four of the five Salmonella pathogenicity islands (SPIs) have been characterized in detail for Salmonella enterica serovar Typhimurium, and the fifth has been characterized for Salmonella enterica serovar Dublin, there have been limited studies to examine them in detail in a range of pathogenic serovars of S. enterica. The aim of this study was to examine these regions, shown to be crucial in virulence, in pathogenic serovars to identify any major deletions or insertions that may explain variation in virulence and provide further understanding of the elements involved in the evolution of these regions. Multiple strains of each of the 13 serovars were compared by Southern blot hybridization using a series of probes that together encompassed the full length of all five SPIs. With the exception of serovar Typhimurium, all strains of the same serovar were identical in all five SPIs. Those serovars that differed from serovar Typhimurium in SPI-1 to SPI-4 and from serovar Dublin in SPI-5 were examined in more detail in the variant regions by PCR, and restriction endonuclease digestion and/or DNA sequencing. While most variation in hybridization patterns was attributable to loss or gain of single restriction endonuclease cleavage sites, three regions, in SPI-1, SPI-3, and SPI-5, had differences due to major insertions or deletions. In SPI-1 the avrA gene was replaced by a 200-base fragment in three serovars, as reported previously. In SPI 5, two serovars had acquired an insertion with similarity to the pagJ and pagK genes between pipC and pipD. In SPI-3 the genes sugR and rhuM were deleted in most serovars and in some were replaced by sequences that were very similar to either the Escherichia coli fimbrial operon, flanked by two distinct insertion sequence elements, or to the E. coli retron phage PhiR73. The distribution of these differences suggests that there have been a number of relatively recent horizontal transfers of genes into S. enterica and that in some cases the same event has occurred in multiple lineages of S. enterica. Thus, it seems that insertion sequences and retron phages are likely to be involved in continuing evolution of the pathogenicity islands of pathogenic Salmonella serovars. PMID- 12775701 TI - Electron microscopic analysis of membrane assemblies formed by the bacterial chemotaxis receptor Tsr. AB - The serine receptor (Tsr) from Escherichia coli is representative of a large family of transmembrane receptor proteins that mediate bacterial chemotaxis by influencing cell motility through signal transduction pathways. Tsr and other chemotaxis receptors form patches in the inner membrane that are often localized at the poles of the bacteria. In an effort to understand the structural constraints that dictate the packing of receptors in the plane of the membrane, we have used electron microscopy to examine ordered assemblies of Tsr in membrane extracts isolated from cells engineered to overproduce the receptor. Three types of assemblies were observed: ring-like "micelles" with a radial arrangement of receptor subunits, two-dimensional crystalline arrays with approximate hexagonal symmetry, and "zippers," which are receptor bilayers that result from the antiparallel interdigitation of cytoplasmic domains. The registration among Tsr molecules in the micelle and zipper assemblies was sufficient for identification of the receptor domains and for determination of their contributions to the total receptor length. The overall result of this analysis is compatible with an atomic model of the receptor dimer that was constructed primarily from the X-ray crystal structures of the periplasmic and cytoplasmic domains. Significantly, the micelle and zipper structures were also observed in fixed, cryosectioned cells expressing the Tsr receptor at high abundance, suggesting that the modes of Tsr assembly found in vitro are relevant to the situation in the cell. PMID- 12775702 TI - Antisense RNA downregulation of coenzyme A transferase combined with alcohol aldehyde dehydrogenase overexpression leads to predominantly alcohologenic Clostridium acetobutylicum fermentations. AB - Plasmid pAADB1 for the overexpression of the alcohol-aldehyde dehydrogenase (aad) gene and downregulation of the coenzyme A transferase (CoAT) using antisense RNA (asRNA) against ctfB (the second CoAT gene on the polycistronic aad-ctfA-ctfB message) was used in order to increase the butanol/acetone ratio of Clostridium acetobutylicum ATCC 824 fermentations. Acetone and butanol levels were drastically reduced in 824(pCTFB1AS) (expresses only an asRNA against ctfB) compared to 824(pSOS95del) (plasmid control). Compared to strain 824(pCTFB1AS), 824(pAADB1) fermentations exhibited two profound differences. First, butanol levels were ca. 2.8-fold higher in 824(pAADB1) and restored back to plasmid control levels, thus supporting the hypothesis that asRNA downregulation of ctfB leads to degradation of the whole aad-ctfA-ctfB transcript. Second, ethanol titers in 824(pAADB1) were ca. 23-fold higher and the highest (ca. 200 mM) ever reported in C. acetobutylicum. Western blot analysis confirmed that CoAT was downregulated in 824(pAADB1) at nearly the same levels as in strain 824(pCTFB1AS). Butyrate depletion in 824(pAADB1) fermentations suggested that butyryl-CoA was limiting butanol production in 824(pAADB1). This was confirmed by exogenously adding butyric acid to 824(pAADB1) fermentations to increase the butanol/ethanol ratio. DNA microarray analysis showed that aad overexpression profoundly affects the large-scale transcriptional program of the cells. Several classes of genes were differentially expressed [strain 824(pAADB1) versus strain 824(pCTFB1AS)], including genes of the stress response, sporulation, and chemotaxis. The expression patterns of the CoAT genes (ctfA and ctfB) and aad were consistent with the overexpression of aad and asRNA downregulation of ctfB. PMID- 12775703 TI - Oxidative stress in Synechococcus sp. strain PCC 7942: various mechanisms for H2O2 detoxification with different physiological roles. AB - This study focuses on the mechanisms for hydrogen peroxide detoxification in Synechococcus sp. strain PCC 7942. To gain better understanding of the role of different routes of hydrogen peroxide detoxification, we inactivated TplA (thioredoxin-peroxidase-like), which we recently identified. In addition, we inactivated the gene encoding catalase-peroxidase and examined the ability to detoxify H(2)O(2) and to survive oxidative stress in both of the single mutants and in the double mutant. Surprisingly, we observed that the double mutant survived H(2)O(2) concentrations that the single catalase-peroxidase mutant could not tolerate. This phenotype correlated with an increased ability of the double mutant to detoxify externally added H(2)O(2) compared to the catalase-peroxidase mutant. Therefore, our studies suggested the existence of a hydrogen peroxide detoxification activity in addition to catalase-peroxidase and thioredoxin peroxidase. The rate of detoxification of externally added H(2)O(2) was similar in the wild-type and the TplA mutant cells, suggesting that, under these conditions, catalase-peroxidase activity was essential for this process and TplA was dispensable. However, during excessive radiation, conditions under which the cell might experience oxidative stress, TplA appears to be essential for growth, and cells lacking it cannot compete with the wild-type strain. Overall, these studies suggested different physiological roles for various cellular hydrogen peroxide detoxification mechanisms in Synechococcus sp. strain PCC 7942. PMID- 12775704 TI - Trigger factor-mediated prolyl isomerization influences maturation of the Streptococcus pyogenes cysteine protease. AB - Trigger factor, a ribosome-associated chaperone and peptidyl-prolyl cis-trans isomerase (PPIase), is essential for the secretion and maturation of the cysteine protease of the pathogenic gram-positive bacterium Streptococcus pyogenes. In the absence of trigger factor, the nascent protease polypeptide is not targeted to the secretory pathway. Some partial-function mutations restore targeting. However, the secreted protease does not efficiently mature into an enzymatically active form, suggesting that trigger factor has an additional role in protease biogenesis. Here, we show that, while not required for targeting, the PPIase activity of trigger factor is essential for maturation of the protease following its secretion from the bacterial cell. Site-specific mutations introduced into ropA, the gene which encodes trigger factor in S. pyogenes, produced mutant proteins deficient in PPIase activity. When these mutant alleles were used to replace the wild-type gene on the streptococcal chromosome, analysis of protease biogenesis revealed that, although the protease was secreted normally, it did not efficiently mature to an active form. Furthermore, mutation of a single proline residue in the protease prodomain suppressed the requirement for PPIase activity, suggesting that this residue is the target of trigger factor. These data support a model in which trigger factor-mediated prolyl isomerization influences the conformation of the prodomain, which in turn directs the protease into one of several alternative folding pathways. PMID- 12775705 TI - Involvement of cyclic AMP (cAMP) and cAMP receptor protein in anaerobic respiration of Shewanella oneidensis. AB - Shewanella oneidensis is a metal reducer that can use several terminal electron acceptors for anaerobic respiration, including fumarate, nitrate, dimethyl sulfoxide (DMSO), trimethylamine N-oxide (TMAO), nitrite, and insoluble iron and manganese oxides. Two S. oneidensis mutants, SR-558 and SR-559, with Tn5 insertions in crp, were isolated and analyzed. Both mutants were deficient in Fe(III) and Mn(IV) reduction. They were also deficient in anaerobic growth with, and reduction of, nitrate, fumarate, and DMSO. Although nitrite reductase activity was not affected by the crp mutation, the mutants failed to grow with nitrite as a terminal electron acceptor. This growth deficiency may be due to the observed loss of cytochromes c in the mutants. In contrast, TMAO reduction and growth were not affected by loss of cyclic AMP (cAMP) receptor protein (CRP). Fumarate and Fe(III) reductase activities were induced in rich medium by the addition of cAMP to aerobically growing wild-type S. oneidensis. These results indicate that CRP and cAMP play a role in the regulation of anaerobic respiration, in addition to their known roles in catabolite repression and carbon source utilization in other bacteria. PMID- 12775706 TI - Altered substrate selection of the melibiose transporter (MelY) of Enterobacter cloacae involving point mutations in Leu-88, Leu-91, and Ala-182 that confer enhanced maltose transport. AB - We isolated mutants of Escherichia coli HS4006 containing the melibiose-H(+) symporter (MelY) from Enterobacter cloacae that had enhanced fermentation on 1% maltose MacConkey plates. DNA sequencing revealed three site classes of mutations: L-88-P, L-91-P, and A-182-P. The mutants L-88-P and L-91-P had 3.6- and 5.1-fold greater maltose uptake than the wild type and enhanced apparent affinities for maltose. Energy-coupled transport was defective for melibiose accumulation, but detectable maltose accumulation for the mutants indicated that active transport is dependent upon the substrate transported through the carrier. We conclude that the residues Leu-88, Leu-91 (transmembrane segment 3 [TMS-3]), and Ala-182 (TMS-6) of MelY mediate sugar selection. These data represent the first MelY mutations that confer changes in sugar selection. PMID- 12775707 TI - Xenorhabdus nematophila requires an intact iscRSUA-hscBA-fdx operon to colonize Steinernema carpocapsae nematodes. AB - An insertion between iscA and hscB of the Xenorhabdus nematophila iscRSUA-hscBA fdx locus, predicted to encode Fe-S assembly machinery, prevented colonization of Steinernema carpocapsae nematodes. The insertion disrupted cotranscription of iscA and hscB, but did not reduce hscBA expression, suggesting that X. nematophila requires coordinated expression of the isc-hsc-fdx locus for colonization. PMID- 12775708 TI - Enhanced estrogen receptor (ER) alpha, ERBB2, and MAPK signal transduction pathways operate during the adaptation of MCF-7 cells to long term estrogen deprivation. AB - The mechanisms involved in resistance to estrogen deprivation are of major importance for optimal patient therapy and the development of new drugs. Long term culture of MCF-7 cells in estrogen (E2)-depleted medium (long term estrogen deprivation; LTED) results in hypersensitivity to E2 coinciding with elevated levels of estrogen receptor (ER) alpha phosphorylated on Ser118 and MAPK, together with several of its downstream targets associated previously with ERalpha phosphorylation. Our data suggest elevated MAPK activity results from enhanced ERBB2 expression in the LTED cells versus the wild-type (wt), and treatment with the tyrosine kinase inhibitor ZD1839 revealed increased sensitivity in both transcription and proliferation assays. Similarly the MEK inhibitor U0126 decreased transcription and proliferation in the LTED cells and reduced their sensitivity to the proliferative effects of E2, while having no effect on the wt. However, the complete suppression of MAPK activity in the LTED cells did not inhibit ERalpha Ser118 phosphorylation suggesting that ER activity remained ligand-dependant. The LTED cells also expressed elevated levels of insulin-like growth factor-1R, and inhibition of phosphatidylinositol 3-kinase activity with LY294002 reduced basal ERalpha transactivation by 70% in the LTED cells compared with the wt. However, LY294002 had no effect on ERalpha Ser118 phosphorylation. These data suggest that although elevated levels of MAPK occur during LTED and influence the phenotype, this is unlikely to be the sole pathway operating to achieve adaptation. PMID- 12775709 TI - Ankyrin peptide blocks falcipain-2-mediated malaria parasite release from red blood cells. AB - Falcipain-2 (FP-2) is a dual-function protease that cleaves hemoglobin at the early trophozoite stage and erythrocyte membrane ankyrin and protein 4.1 at the late stages of parasite development. FP-2-mediated cleavage of ankyrin and protein 4.1 is postulated to cause membrane instability facilitating parasite release in vivo. To test this hypothesis, here we have determined the precise peptide sequence at the hydrolysis site of ankyrin to develop specific inhibitor(s) of FP-2. Mass spectrometric analysis of the hydrolysis products showed that FP-2-mediated cleavage of ankyrin occurred immediately after arginine 1,210. A 10-mer peptide (ankyrin peptide, AnkP) containing the cleavage site completely inhibited the FP-2 enzyme activity in vitro and abolished all of the known functions of FP-2. To determine the effect of this peptide on the growth and development of P. falciparum, the peptide was delivered into intact parasite infected red blood cells (RBCs) via the Antennapedia homeoprotein internalization domain. Growth and maturation of trophozoites and schizonts was markedly inhibited in the presence of the fused AnkP peptide. <10% of new ring-stage parasites were detected compared with the control sample. Together, our results identify a specific peptide derived from the spectrin-binding domain of ankyrin that blocks late-stage malaria parasite development in RBCs. Confocal microscopy with FP-2-specific antibodies demonstrated the proximity of the enzyme in apposition with the RBC membrane, further corroborating the proposed function of FP-2 in the cleavage of RBC skeletal proteins. PMID- 12775710 TI - The structural organization of sperm chromatin. AB - The packaging of the male haploid genome within the differentiating spermatid nucleus is facilitated by small basic nuclear proteins called protamines. Although the majority of the DNA in human sperm chromatin is bound by these proteins, a small percentage retains a nucleosomal-like component. These histone enriched regions may possess enhanced nuclease sensitivity and have been postulated to designate certain genes involved in early embryogenesis. We have shown previously that the chromatin domain containing the two human protamines PRM1 and PRM2 and the transition protein TNP2 forms a DNase I-sensitive conformation in pachytene spermatocytes, a requisite event prior to the haploid expression of its members in round spermatids (Kramer, J. A, McCarrey, J., Djakiew, D., and Krawetz, S. A. (1998) Development 125, 4749-4755). Interestingly, this configuration persists in mature spermatozoa subsequent to the transcriptional silencing of the locus. It was therefore postulated that the retained, enhanced DNase I-sensitive conformation of the PRM1-->PRM2-->TNP2 domain in human sperm may be preferentially histone-enriched. To address this tenet, we examined the chromatin structure of the human PRM1--> PRM2--> TNP2 domain using a PCR-based assay. The results show that this retained, enhanced DNase I sensitive domain reflects an enrichment of histones at discrete regions across the locus. In addition, a similar examination of other genes and repetitive sequences suggests the non-random distribution of histones and protamines within the sperm nucleus. A discussion of these results and their functional significance is presented. PMID- 12775711 TI - Structure and ligand-induced conformational change of the 39-kDa glycoprotein from human articular chondrocytes. AB - The 39-kDa human cartilage glycoprotein (HCGP39), a member of a novel family of chitinase-like lectins (Chilectins), is overexpressed in articular chondrocytes and certain cancers. Proposed functions of this protein include a role in connective tissue remodeling and defense against pathogens. Similar to other Chi lectins, HCGP39 promotes the growth of connective tissue cells. The ability of HCGP39 to activate cytoplasmic signaling pathways suggests the presence of a ligand for this protein at the cell surface. There is currently no information regarding the identity of any physiological or pathological ligands of the Chi lectins or the nature of the protein-ligand interaction. Here, we show that HCGP39 is able to bind chitooligosaccharides with micromolar affinity. Crystal structures of the native protein and a complex with GlcNAc8 show that the ligand is bound in identical fashion to family 18 chitinases. However, unlike the chitinases, binding of the oligosaccharide ligand to HCGP39 induces a large conformational change. Thus, HCGP39 could be a lectin that binds chitin-like oligosaccharide ligands and possibly plays a role in innate responses to chitinous pathogens, such as fungi and nematodes. PMID- 12775712 TI - Characterization of multiple signaling pathways of insulin in the regulation of vascular endothelial growth factor expression in vascular cells and angiogenesis. AB - The effects of insulin on vascular endothelial growth factor (VEGF) expression in cultured vascular cells and in angiogenesis were characterized. Insulin increased VEGF mRNA levels in mouse aortic smooth muscle cells from 10(-9) to 10(-7) m with an initial peak of 3.7-fold increases at 1 h and a second peak of 2.8-fold after 12 h. The first peak of VEGF expression was inhibited by LY294002, an inhibitor of phosphatidylinositol (PI) 3-kinase, and by the overexpression of dominant negative forms of p85 subunit of PI 3-kinase or Akt. Inhibitors of MEK kinase, PD98059, or overexpression of dominant negative forms of Ras was ineffective. In contrast, the chronic effect of insulin on VEGF expression was partially inhibited by both LY294002 or PD98059 as well as by the overexpression of dominant negatives of PI 3-kinase or Ras. The importance of PI 3-kinase-Akt pathway on VEGF expression was confirmed in mouse aortic smooth muscle cells isolated from insulin receptor substrate -1 knockout (IRS-1-/-) mice that showed parallel reductions of 46-49% in insulin-stimulated VEGF expression and PI 3 kinase-Akt activation. Insulin-induced activation of PI 3-kinase-Akt on hypoxia induced VEGF expression and neovascularization was reduced by 40% in the retina of neonatal hypoxia model using IRS-1-/- mice. Thus, unlike other cells, insulin can regulate VEGF expression by both IRS-1/PI 3-kinase-Akt cascade and Ras-MAPK pathways in aortic smooth muscle cells. The in vivo results provide direct evidence that insulin can modulate hypoxia-induced angiogenesis via reduction in VEGF expression in vivo. PMID- 12775713 TI - Specific binding of dehydroepiandrosterone to the N terminus of the microtubule associated protein MAP2. AB - The effect of neurosteroids is mediated through their membrane or nuclear receptors. However, no dehydroepiandrosterone (DHEA)-specific receptors have been evidenced so far in the brain. In this paper, we showed by isothermal titration calorimetry that the DHEA specifically binds to the dendritic brain microtubule associated protein MAP2C with an association constant of 2.7 x 10(7) m-1 and at a molar ratio of 1:1. By partial tryptic digestions and mass spectrometry analysis, we found that the binding involved the N-terminal region of MAP2C. Interestingly, MAP2C displays homologies with 17 beta-hydroxysteroid dehydrogenase 1, an enzyme required for estrogen synthesis. Based on these sequence homologies and on the x ray structure of the DHEA-binding pocket of 17 beta-hydroxysteroid dehydrogenase 1, we modeled the complex of DHEA with MAP2C. The binding of DHEA to MAP2C involved specific hydrogen bonds that orient the steroid into the pocket. This work suggests that DHEA can directly influence brain plasticity via MAP2C binding. It opens interesting ways for understanding the role of DHEA in the brain. PMID- 12775714 TI - Functional, persistent, and extended liver to pancreas transdifferentiation. AB - Pancreatic and duodenal homeobox gene-1 (PDX-1) regulates pancreas development during embryogenesis, whereas in the adult it controls beta-cell function. Here we analyze whether PDX-1 functions as a pancreatic differentiation factor and a bona fide master regulator when ectopically expressed in mature fully differentiated liver in vivo. By ectopic and transient PDX-1 expression in liver in vivo, using the first generation recombinant adenoviruses, we demonstrate that PDX-1 induces in liver a wide repertoire of both exocrine and endocrine pancreatic gene expression. Moreover, PDX-1 induces its own expression (auto induction), which in turn may explain the long lasting nature of the "liver to pancreas" transdifferentiation. Insulin as well glucagon-producing cells are mainly located in the proximity of hepatic central veins, possibly allowing direct hormone release into the bloodstream, without affecting normal hepatic function. Importantly, we demonstrate that hepatic insulin production triggered by Ad-CMV-PDX-1 recombinant adenovirus administration is functional and prevents streptozotocin-induced hyperglycemia in Balb/c mice even 8 months after the initial treatment. We conclude that PDX-1 plays an important instructive role in pancreas differentiation, not only from primitive gut endoderm but also from mature liver. Transconversion of liver to pancreas may serve as a novel approach for generating endocrine-pancreatic tissue that can replace malfunctioning beta cells in diabetics. PMID- 12775715 TI - Biosynthetic processing of cathepsins and lysosomal degradation are abolished in asparaginyl endopeptidase-deficient mice. AB - Asparaginyl endopeptidase (AEP)/legumain, an asparagine-specific cysteine proteinase in animals, is an ortholog of plant vacuolar processing enzyme (VPE), which processes the exposed asparagine residues of various vacuolar proteins. In search for its physiological role in mammals, here we generated and characterized AEP-deficient mice. Although their body weights were significantly reduced, they were normally born and fertile. In the wild-type kidney where the expression of AEP was exceedingly high among various organs, the localization of AEP was mainly found in the lamp-2-positive late endosomes in the apical region of the proximal tubule cells. In these cells of AEP-deficient mice, the lamp-2-positive membrane structures were found to be greatly enlarged. These aberrant lysosomes, merged with the late endosomes, accumulated electron-dense and membranous materials. Furthermore, the processing of the lysosomal proteases, cathepsins B, H, and L, from the single-chain forms into the two-chain forms was completely defected in the deficient mice. Thus, the AEP deficiency caused the accumulation of macromolecules in the lysosomes, highlighting a pivotal role of AEP in the endosomal/lysosomal degradation system. PMID- 12775716 TI - cGMP-dependent protein kinase type II regulates basal level of aldosterone production by zona glomerulosa cells without increasing expression of the steroidogenic acute regulatory protein gene. AB - The renin-angiotensin-aldosterone system plays a pivotal role in the regulation of salt and water homeostasis. Here, we demonstrate the expression and functional role of cGMP-dependent protein kinases (PKGs) in rat adrenal cortex. Expression of PKG II is restricted to adrenal zona glomerulosa (ZG) cells, whereas PKG I is localized to the adrenal capsule and blood vessels. Activation of the aldosterone system by a low sodium diet up-regulated the expression of PKG II, however, it did not change PKG I expression in adrenal cortex. Both, activation of PKG II in isolated ZG cell and adenoviral gene transfer of wild type PKG II into ZG cells enhanced aldosterone production. In contrast, inhibition of PKG II as well as infection with a PKG II catalytically inactive mutant had an inhibitory effect on aldosterone production. Steroidogenic acute regulatory (StAR) protein that regulates the rate-limiting step in steroidogenesis is a new substrate for PKG II and can be phosphorylated by PKG II in vitro at serine 55/56 and serine 99. Stimulation of aldosterone production by PKG II in contrast to stimulation by PKA did not activate StAR gene expression in ZG cells. The results presented indicate that PKG II activity in ZG cells is important for maintaining basal aldosterone production. PMID- 12775717 TI - Persistent protease-activated receptor 4 signaling mediates thrombin-induced microglial activation. AB - We have previously reported that thrombin, the ultimate serine protease in the coagulation cascades, is a proinflammatory agent that causes proliferation and activation of brain microglial cells. However, participation of its principal receptor, the protease-activated receptor 1 (PAR1) appears to be limited to promoting microglial proliferation and not induction of inflammatory mediators. In the present study, we now report that thrombin action in promoting inflammatory mediators from brain microglia is mediated through another thrombin receptor, PAR4. Here we show that the PAR4 agonist peptide (PAR4AP, GYPGKF), but not the PAR1AP (TRAP, SFLLRN), induced tumor necrosis factor-alpha (TNF-alpha) production not only in cultured murine microglial cells in vitro but also in rat cortex in vivo. Down-regulation of PAR4 expression in microglial cultures by a specific antisense, but not a sense, oligonucleotide reduced PAR4AP-induced TNF alpha. Mechanistic studies indicated that, in comparison with PAR1 signaling, prolonged increase of [Ca2+]i and phosphorylation of p44/42 mitogen-activated protein kinases, as well as NFkappaB activation may be responsible for PAR4AP induced TNF-alpha production in microglia. Taken together, these results demonstrate that PAR4 activation mediates the potentially detrimental effects of thrombin on microglia, implying that perspectives of exploiting PAR1 as a potential anti-inflammatory target should be shifted toward PAR4 as a much more specific therapeutic target in brain inflammatory conditions associated with neurotrauma and neurodegenerations. PMID- 12775718 TI - ADAMTS-13 metalloprotease interacts with the endothelial cell-derived ultra-large von Willebrand factor. AB - Thrombotic thrombocytopenic purpura is caused by congenital or acquired deficiency of ADAMTS-13, a metalloprotease that cleaves the endothelium-derived ultra-large multimers of von Willebrand factor (ULVWF). The proteolysis converts hyper-reactive and thrombogenic ULVWF into smaller and less adhesive plasma forms. Activity of ADAMTS-13 is usually measured in a static system under non physiological conditions that require protein denaturation and prolonged incubation. We have demonstrated previously that ULVWF multimers, upon release from endothelial cells, form platelet-decorated string-like structures that are rapidly cleaved by ADAMTS-13. Here we report the direct interaction between ADAMTS-13 and VWF under both static and flowing conditions. ADAMTS-13-coated beads adhered to both immobilized VWF and ULVWF strings presented by stimulated endothelial cells. These beads adhered to VWF under both venous (2.5 dynes/cm2) and arterial (30 dynes/cm2) shear stresses. We then demonstrated that ADAMTS-13 beads adhered to immobilized recombinant VWF-A1 and -A3 domains, but soluble metalloprotease bound preferentially to the A3 domain, suggesting that the VWF A3 domain may be the primary docking site for the metalloprotease. We suggest that tensile stresses imposed by fluid shear stretch endothelial bound ULVWF multimers to expose binding sites within the A domains for circulating ADAMTS-13. The bound enzyme then cleaves within the A2 domain that lies in close proximity and releases smaller VWF multimers into the plasma. Once released, these cleaved VWF fragments become inaccessible for the metalloprotease to prevent further cleavage. PMID- 12775719 TI - CARD6 is a modulator of NF-kappa B activation by Nod1- and Cardiak-mediated pathways. AB - We cloned a novel cDNA derived from the CARD6 gene locus on chromosome 5p12 of 311 amino acids in length. By immunoprecipitation we detected specific binding of this CARD6-encoding protein to Nod1 (CARD4), Cardiak (Rip2/Rick), NAC (NALP1/DEFCAP/CARD7), and TUCAN (CARD8/Cardinal/NDPP/Dakar), caspase recruitment domain (CARD)-containing proteins implicated in NF-kappa B and caspase-1 activation but not to other CARD family proteins. Cardiak and Nod1 (but not other CARD proteins) also exhibited opposing effects on CARD6 protein phosphorylation and expression, providing further evidence of functional interactions among these proteins in cells. In transfection experiments, the CARD6 protein suppressed NF kappa B induction by Nod1 or Cardiak but did not interfere with NF-kappa B activation by the CARD-containing adapter protein Bcl10 or the cytokine tumor necrosis factor-alpha, demonstrating specificity of CARD6 for Nod-1 and Cardiak dependent pathways. In contrast to its effects on Nod1- and Cardiak-dependent NF kappa B activation, CARD6 did not interfere with caspase-1-dependent interleukin 1 beta secretion induced by Cardiak or Nod1. CARD6 also did not affect caspase activation and apoptosis induced by overexpression of Fas, Bax, or other pro apoptotic stimuli. Thus, CARD6 represents a selective modulator of NF-kappa B activation by Cardiak and Nod1, adding to the repertoire of CARD-family proteins implicated in inflammatory responses and innate immunity. PMID- 12775720 TI - Platelet endothelial cell adhesion molecule-1 (PECAM-1) inhibits low density lipoprotein-induced signaling in platelets. AB - At physiological concentrations, low density lipoprotein (LDL) increases the sensitivity of platelets to aggregation- and secretion-inducing agents without acting as an independent activator of platelet functions. LDL sensitizes platelets by inducing a transient activation of p38MAPK, a Ser/Thr kinase that is activated by the simultaneous phosphorylation of Thr180 and Tyr182 and is an upstream regulator of cytosolic phospholipase A2 (cPLA2). A similar transient phosphorylation of p38MAPK is induced by a peptide mimicking amino acids 3359 3369 in apoB100 called the B-site. Here we report that the transient nature of p38MAPK activation is caused by platelet endothelial cell adhesion molecule 1 (PECAM-1), a receptor with an immunoreceptor tyrosine-based inhibitory motif. PECAM-1 activation by cross-linking induces tyrosine phosphorylation of PECAM-1 and a fall in phosphorylated p38MAPK and cPLA2. Interestingly, LDL and the B-site peptide also induce tyrosine phosphorylation of PECAM-1, and studies with immunoprecipitates indicate the involvement of c-Src. Inhibition of the Ser/Thr phosphatases PP1/PP2A (okadaic acid) makes the transient p38MAPK activation by LDL and the B-site peptide persistent. Inhibition of Tyr-phosphatases (vanadate) increases Tyr-phosphorylated PECAM-1 and blocks the activation of p38MAPK. Together, these findings suggest that, following a first phase in which LDL, through its B-site, phosphorylates and thereby activates p38MAPK, a second phase is initiated in which LDL activates PECAM-1 and induces dephosphorylation of p38MAPK via activation of the Ser/Thr phosphatases PP1/PP2A. PMID- 12775721 TI - Resistance to tumor necrosis factor-alpha (TNF-alpha)-induced apoptosis in rat hepatoma cells expressing TNF-alpha is linked to low antioxidant enzyme expression. AB - In order to study the mechanisms of resistance to tumor necrosis factor-alpha (TNF-alpha), we have constructed two stable transfectants producing TNF-alpha (Yv12-2 and Yv13-44) from the rat hepatoma H4IIE cell, which does not produce TNF alpha. H4IIE cells were highly sensitive to apoptosis induced by TNF-alpha, whereas Yv2-12 and Yv13-44 cells were resistant. Manganous superoxide dismutase was not up-regulated in Yv2-12 and Yv13-44 cells and was unresponsive to induction by exogenous TNF-alpha and by H2O2 in H4IIE cells and in the transfectants. Catalase expression and activity were lower in Yv2-12 and Yv13-44 cells than in H4IIE cells; furthermore, the transfectants were more susceptible to H2O2. Treatment with exogenous TNF-alpha down-regulated catalase in H4IIE cells but not in Yv2-12 and Yv13-44 cells. Treatment of H4IIE cells with the catalase inhibitor 3-amino-1,2,4-triazole rendered them resistant to exogenous TNF-alpha. These data suggest a causal relationship between resistance to TNF alpha and low catalase activity. Expression of copper and zinc containing superoxide dismutase was also decreased, whereas expression of glutathione peroxidase-1 was unchanged in Yv2-12 and Yv13-44 cells. Data from a microarray point to a down-regulation of genes in the resistant clones that code for antioxidative proteins and proteins involved in glutathione synthesis and function. We assume that a prooxidant signal linked to the down-regulation of antioxidant defense may be associated with resistance to apoptosis induced by TNF alpha. PMID- 12775722 TI - Differential requirement of SWI/SNF for androgen receptor activity. AB - The androgen receptor (AR) is a ligand-dependent transcription factor whose activity is required for prostate cancer proliferation. Because ablation of AR activity is a critical goal of prostate cancer therapy, much emphasis has been placed on understanding the accessory proteins that regulate AR function in the prostate. Several co-activators have been shown to be required for full AR activity, including histone acetyl-transferases and TRAP/mediator complexes. SWI/SNF comprises a family of large, multisubunit complexes present in the cell, which contain one of two core ATPases required for nucleosome re-positioning, BRG1 or hBRM. We investigated the specific requirement of the SWI/SNF core ATPases for AR function. Using cells deficient in both BRG1 and hBRM, we show that activation of one AR target promoter, prostate-specific antigen (PSA), requires SWI/SNF chromatin remodeling for activity. A second AR target promoter, probasin, maintained a low level of activation in the absence of SWI/SNF. AR stimulation on the probasin core promoter could be partially induced with BRG1, but hBRM strongly stimulated AR activity. The PSA promoter was only induced by the restoration of hBRM. In contrast, ligand-dependent activation of the estrogen receptor was equally stimulated by BRG1 or hBRM. We demonstrate that the addition of a known enhancer region to the core PSA promoter bypasses the requirement for SWI/SNF on the PSA promoter, indicating that elements upstream of specific proximal promoters can impact the influence of the SWI/SNF complex on target gene activation. Addition of the enhancer to the probasin core promoter failed to impact the SWI/SNF requirement. In summary, SWI/SNF function potently regulates core AR target gene promoter activation, with a preference for hBRM-containing complexes. These studies highlight a role for the enhancer in altering the impact of SWI/SNF action and suggest a disparity in AR target genes for SWI/SNF requirement. PMID- 12775723 TI - Acylation state of the phosphatidylinositol hexamannosides from Mycobacterium bovis bacillus Calmette Guerin and mycobacterium tuberculosis H37Rv and its implication in Toll-like receptor response. AB - The dimannoside (PIM2) and hexamannoside (PIM6) phosphatidyl-myo-inositol mannosides are the two most abundant classes of PIM found in Mycobacterium bovis bacillus Calmette Guerin, Mycobacterium tuberculosis H37Rv, and Mycobacterium smegmatis 607. Recently, these long known molecules received a renewed interest due to the fact that PIM2 constitute the anchor motif of an important constituent of the mycobacterial cell wall, the lipoarabinomannans (LAM), and that both LAM (phosphoinositol-capped LAM) and PIM are agonists of Toll-like receptor 2 (TLR2), a pattern recognition receptor involved in innate immunity. Due to the biological importance of these molecules, the chemical structure of PIM was revisited. The structure of PIM2 was recently published (Gilleron, M., Ronet, C., Mempel, M., Monsarrat, B., Gachelin, G., and Puzo, G. (2001) J. Biol. Chem. 276, 34896 34904). Here we report the purification and molecular characterization of PIM6 in their native form. For the first time, four acyl forms of this molecule have been purified, using hydrophobic interaction chromatography. Mono- to tetra-acylated molecules were identified in M. bovis bacillus Calmette Guerin, M. tuberculosis H37Rv, and M. smegmatis 607 using a sophisticated combination of analytical tools, including matrix-assisted laser desorption/ionization-time of flight-mass spectrometry and two-dimensional homo- and heteronuclear NMR spectroscopy. These experiments revealed that the major acyl forms are similar to the ones described for PIM2. Finally, we show that PIM6, like PIM2, activate primary macrophages to secrete TNF-alpha through TLR2, irrespective of their acylation pattern, and that they signal through the adaptor MyD88. PMID- 12775724 TI - RLIP, an effector of the Ral GTPases, is a platform for Cdk1 to phosphorylate epsin during the switch off of endocytosis in mitosis. AB - The Ral signaling pathway is critically involved in Ras-dependent oncogenesis. One of its key actors, RLIP/RalBP1, which participates in receptor endocytosis during interphase, is also involved in mitotic processes when endocytosis is switched off. During mitosis, RLIP76 is located on the duplicated centrosomes and is required for their proper separation and movement to the poles. We have looked for actors that associate with RLIP during mitosis. We show here that RLIP/RalBP1 interacts with an active p34cdc2.cyclinB1 (cdk1) enzyme and that this interaction is crucial for the mitotic phosphorylation of Epsin that, once phosphorylated, is no longer competent for endocytosis. We show also that this latter phosphorylation is dependent on Ral signaling. We propose that RLIP/RalBP1 is used as a platform by the mitotic cdk1 to facilitate the phosphorylation of Epsin, which makes Epsin incompetent for endocytosis during mitosis, when endocytosis is switched off. PMID- 12775725 TI - Multiple, independently regulated pathways of cholesterol transport across the intestinal epithelial cells. AB - The present study provides a new understanding about the mechanisms involved in cholesterol absorption by the intestinal cells. Contrary to general belief, our data show that newly absorbed cholesterol is neither immediately available for secretion with apoB lipoproteins nor exclusively secreted as part of chylomicrons. Based on our data, cholesterol transport by enterocytes can be broadly classified into two independently modulated, apoB-dependent and independent, pathways. Cholesterol secretion by the apoB-dependent pathway is induced by oleic acid, is repressed by microsomal triglyceride transfer protein inhibitors, and occurs only with larger apoB-containing lipoproteins. ApoB independent pathways do not require microsomal triglyceride transfer protein and involve efflux mediated by ABCA1, high density lipoprotein assembly, and possibly other unknown mechanisms. There are at least two different metabolic pools of cholesterol. The newly absorbed and pre-absorbed cholesterol are preferentially secreted via apoB-independent and apoB-dependent pathways, respectively. In contrast to compartmentalization for secretion, these two metabolic pools are equally accessible for cellular esterification. The esterified cholesterol is mainly secreted by the apoB-dependent pathway, whereas both the pathways are involved in the secretion of free cholesterol. Thus, enterocytes transport exogenous cholesterol by several independently regulated pathways raising the possibility that targeting of apoB-independent pathways may result in selective inhibition of cholesterol transport without affecting triglyceride transport. PMID- 12775726 TI - Malondialdehyde, a product of lipid peroxidation, is mutagenic in human cells. AB - Malondialdehyde (MDA) is an endogenous genotoxic product of enzymatic and oxygen radical-induced lipid peroxidation whose adducts are known to exist in DNA isolated from healthy human beings. To evaluate the mutagenic potential of MDA in human cells, we reacted MDA with pSP189 shuttle vector DNA and then transfected them into human fibroblasts for replication. MDA induced up to a 15-fold increase in mutation frequency in the supF reporter gene compared with untreated DNA. Sequence analysis revealed that the majority of MDA-induced mutations occurred at GC base pairs. The most frequent mutations were large insertions and deletions, but base pair substitutions were also detected. MDA-induced mutations were completely abolished when the adducted shuttle vector was replicated in cells lacking nucleotide excision repair. MDA induction of large deletions and the apparent requirement for nucleotide excision repair suggested the possible involvement of a DNA interstrand cross-link as a premutagenic lesion. Indeed, MDA formed interstrand cross-links in duplex plasmids and oligonucleotides. Substrates containing the sequence 5'-d(CG) were preferentially cross-linked, consistent with the observation of base pair substitutions in 5'-d(CG) sites in the MDA-induced mutation spectrum. These experiments provide biological and biochemical evidence for the existence of MDA-induced DNA interstrand cross-links that could result from endogenous oxidative stress and likely have potent biological effects. PMID- 12775727 TI - Oxaliplatin for colorectal cancer in the United States: better late than never. PMID- 12775728 TI - Molecular oncodiagnostics: where we are and where we need to go. PMID- 12775729 TI - Cyclin D1 expression in the field of exposure: another piece in the molecular Auerbach puzzle. PMID- 12775730 TI - Superiority of oxaliplatin and fluorouracil-leucovorin compared with either therapy alone in patients with progressive colorectal cancer after irinotecan and fluorouracil-leucovorin: interim results of a phase III trial. AB - PURPOSE: In North America, no effective therapy has been available for patients with progressive metastatic colorectal cancer after front-line treatment with irinotecan, bolus fluorouracil (FU), and leucovorin (IFL). PATIENTS AND METHODS: Patients with metastatic colorectal cancer who progressed after IFL therapy were randomly assigned to bolus and infusional FU and leucovorin (LV5FU2), single agent oxaliplatin, or the combination (FOLFOX4). This planned interim analysis evaluated objective response rate (RR), time to tumor progression (TTP), and alleviation of tumor-related symptoms (TRS) in an initial cohort of patients. RESULTS: Between November 2000 and September 2001, 463 patients from 120 sites in North America were randomly assigned to treatment. FOLFOX4 proved superior to LV5FU2 in all measures of clinical efficacy. Objective RRs determined by an independent radiology panel were 9.9% for FOLFOX4 versus 0% for LV5FU2 (Fisher's exact test, P <.0001). Median TTP was 4.6 months for FOLFOX4 versus 2.7 months for LV5FU2 (two-sided, stratified log-rank test, P <.0001). Relief of TRS occurred in 33% of patients treated with FOLFOX4 versus 12% of patients treated with LVFU2 (chi2 test, P <.001). Single-agent oxaliplatin was not superior to LV5FU2 in any measure of efficacy. Patients treated with FOLFOX4 experienced a higher incidence of clinically significant toxicities than patients treated with LV5FU2, but these toxicities were predictable and did not result in a higher rate of treatment discontinuation or 60-day mortality rate. CONCLUSION: For patients with metastatic colorectal cancer, second-line treatment with FOLFOX4 is superior to treatment with LVFU2 in terms of RR, TTP, and relief of TRS. PMID- 12775731 TI - Asian ethnicity-related differences in gastric cancer presentation and outcome among patients treated at a canadian cancer center. AB - PURPOSE: Differences in stage-stratified survival have been reported between Asian and Western populations with gastric cancer. This study examines differences in presentation and outcomes among Asian and non-Asian patients evaluated and treated at a Canadian institution. PATIENTS AND METHODS: We reviewed 2,043 patients (159 Asians and 1,884 non-Asians) with gastric adenocarcinoma treated between 1978 and 1997. Overall survival was examined by the Kaplan-Meier method, and multivariable analysis by Cox proportional hazards was used to identify whether Asian ethnicity had independent prognostic significance for survival. RESULTS: Median survival was 13.1 months for Asians and 11.1 months for non-Asians (P =.0016). Asian patients were younger and had a greater proportion of signet ring cell histology but were less likely to have proximal disease. Signet ring cell histology did not adversely affect survival. By multivariable analysis, proximal location, poor differentiation, and extent of disease were independently associated with worse survival. Survival was improved with curative resection, palliative resection, and palliative chemotherapy. Asian ethnicity was not independently associated with survival (hazard ratio, 0.89; 95% confidence interval, 0.74 to 1.08). Although a similar proportion of patients underwent curative resection, an interaction was observed between Asian ethnicity and efficacy of resection, with Asians achieving a greater benefit as compared with non-Asians even when adjusted for age and location. CONCLUSION: The disparity between Eastern and Western gastric cancer survival is not explained by the hypothesis of ethnicity-related differences in tumor biology. Although it is not an independent predictor of survival, Asian ethnicity is associated with distinct characteristics at presentation and more favorable outcomes after curative surgery. PMID- 12775733 TI - Cyclin D1 overexpression in bronchial epithelia of patients with lung cancer is associated with smoking and predicts survival. AB - PURPOSE: Cyclin D1 is overexpressed in almost 60% of resectable non-small-cell lung cancer (NSCLC). In the absence of cyclin D1 gene amplification, overexpression is characterized by allelic imbalanced transcript levels. METHODS: The aims were to study cyclin D1 expression by immunohistochemistry and allelic balance of transcripts in tumor-free bronchial epithelia from patients with resectable NSCLC by using monoclonal antibodies (48 patients and 288 sites), microdissection/reverse transcriptase polymerase chain reaction/restriction fragment length polymorphism analyses (24 patients and 144 sites). Derived data were related to patient characteristics-in particular, smoking habits. RESULTS: In 167 (58%) of 288 sites, cyclin D1 was overexpressed, with cytoplasmic and nuclear sublocalization in 53% and 7% of all sites, respectively. Nuclear overexpression was more frequent in premalignant versus normal or hyperplastic epithelia (55% v 3%; P <.0001). Allele-specific expression imbalances were found in 69 (48%) of 144 sites; in particular, those in which cyclin D1 was overexpressed (P =.004). In 14 (58%) of 24 patients, balanced or imbalanced transcript ratios and degree of expression were consistent at all sites for the same patient, whereas in another 10 patients, transcript balances and cyclin D1 expression patterns varied across the sites. Nuclear cyclin D1 expression in at least one site (14 of 48 patients) was linked to heavy smoking (> 40 pack-years; P =.02) and shorter overall survival (P =.01). CONCLUSION: Allele-specific, probably damage-driven, deregulation of the cyclin D1 gene may precede and perhaps facilitate the spread of preneoplastic clones across the bronchial epithelial surface in a significant number of patients. Cyclin D1 expression at multiple bronchial sites may identify a subgroup of heavy-smoking patients with poor outcome. PMID- 12775732 TI - Quality assessment of genetic markers used for therapy stratification. AB - PURPOSE: Therapy stratification based on genetic markers is becoming increasingly important, which makes commitment to the highest possible reliability of the involved markers mandatory. In neuroblastic tumors, amplification of the MYCN gene is an unequivocal marker that indicates aggressive tumor behavior and is consequently used for therapy stratification. To guarantee reliable and standardized quality of genetic features, a quality-assessment study was initiated by the European Neuroblastoma Quality Assessment (ENQUA; connected to International Society of Pediatric Oncology) Group. MATERIALS AND METHODS: One hundred thirty-seven coded specimens from 17 tumors were analyzed in 11 European national/regional reference laboratories using molecular techniques, in situ hybridization, and flow and image cytometry. Tumor samples with divergent results were re-evaluated. RESULTS: Three hundred fifty-two investigations were performed, which resulted in 23 divergent findings, 17 of which were judged as errors after re-evaluation. MYCN analyses determined by Southern blot and in situ hybridization led to 3.7% and 4% of errors, respectively. Tumor cell content was not indicated in 32% of the samples, and 11% of seemingly correct MYCN results were based on the investigation of normal cells (eg, Schwann cells). Thirty-eight investigations were considered nonassessable. CONCLUSION: This study demonstrated the importance of revealing the difficulties and limitations for each technique and problems in interpreting results, which are crucial for therapeutic decisions. Moreover, it led to the formulation of guidelines that are applicable to all kinds of tumors and that contain the standardization of techniques, including the exact determination of the tumor cell content. Finally, the group has developed a common terminology for molecular-genetic results. PMID- 12775734 TI - Pilot trial of the epidermal growth factor receptor tyrosine kinase inhibitor gefitinib plus carboplatin and paclitaxel in patients with stage IIIB or IV non small-cell lung cancer. AB - PURPOSE: Gefitinib is an oral agent that inhibits the tyrosine kinase of the epidermal growth factor receptor. In phase I trials gefitinib was well tolerated and antitumor activity was seen in pretreated non-small-cell lung cancer (NSCLC) patients. Preclinical studies indicated enhanced effects when gefitnib was added to carboplatin or paclitaxel. This pilot trial combined gefitinib with carboplatin and paclitaxel to define the toxicities of the combination and assess drug-drug interactions in untreated advanced NSCLC patients. PATIENTS AND METHODS: Initially (part 1) patients were randomly assigned to receive intermittent gefitinib with cycle 1 or 2 of chemotherapy. Thereafter (part 2), the highest dose of gefitinib that was given without dose-limiting toxicity (DLT) from part 1 was administered continuously beginning with the first cycle of chemotherapy. Three sequentially enrolled cohorts received gefitinib 250 and 500 mg (intermittently) and 500 mg (continuously). RESULTS: We treated 24 patients; nine patients with 250 mg and 15 patients with 500 mg (nine patients continuous). Two occurrences of DLT were observed. One patient (500 mg, part 1) developed grade 3 rash and another patient (part 2) developed prolonged neutropenia. Steady state gefitinib levels did not affect exposure to chemotherapy. In a limited sample, chemotherapy modestly increased the gefitinib area under concentration time curve at steady-state and minimum steady-state trough concentration. Partial responses were observed in five of 24 patients. The median survival was 8 months. CONCLUSION: The gefitinib with carboplatin and paclitaxel regimen was generally well tolerated and no unanticipated toxicities or clinically relevant pharmacokinetic interactions were observed. Both doses of gefitinib were believed to be safe for further study with chemotherapy. This regimen was thus tested in a completed randomized phase III trial. PMID- 12775735 TI - Phase III study of letrozole versus tamoxifen as first-line therapy of advanced breast cancer in postmenopausal women: analysis of survival and update of efficacy from the International Letrozole Breast Cancer Group. AB - PURPOSE: To analyze overall survival (OS) and update efficacy data for letrozole versus tamoxifen as first-line therapy in postmenopausal women with locally advanced or metastatic breast cancer. PATIENTS AND METHODS: This multicenter phase III trial randomly assigned 916 patients with hormone receptor-positive or unknown tumors letrozole 2.5 mg (n = 458) or tamoxifen 20 mg (n = 458) daily until disease progression. Optional cross-over was permitted at the treating physician's discretion. This report updates efficacy at a median follow-up of 32 months. RESULTS: The superiority of letrozole to tamoxifen was confirmed for time to progression (median, 9.4 v 6.0 months, respectively; P <.0001), time to treatment failure (median, 9 v 5.7 months, respectively; P <.0001), overall objective response rate (32% v 21%, respectively; P =.0002), and overall clinical benefit. Median OS was slightly prolonged for the randomized letrozole arm (34 v 30 months, respectively). Although this difference in OS is not significant, survival was improved in the randomized letrozole arm over the first 2 years of the study. Approximately one half of the patients in each arm crossed over. Total duration of endocrine therapy ("time to chemotherapy") was significantly longer (P =.005) for patients initially on letrozole (median, 16 months) than for patients initially on tamoxifen (median, 9 months). Time to worsening of Karnofsky performance score was significantly delayed with letrozole compared with tamoxifen (P =.001). CONCLUSION: This study documents the superiority of letrozole over tamoxifen in first-line endocrine therapy in postmenopausal women with advanced breast cancer. PMID- 12775736 TI - Topotecan is an active agent in the first-line treatment of metastatic or recurrent endometrial carcinoma: Eastern Cooperative Oncology Group Study E3E93. AB - PURPOSE: To determine the clinical activity and the toxicity profile of the topoisomerase-I inhibitor, topotecan, in women with recurrent or advanced endometrial carcinoma. PATIENTS AND METHODS: A prospective, phase II clinical trial was initiated by the Eastern Cooperative Oncology Group (ECOG). Patients had histologically confirmed advanced or recurrent endometrial carcinoma, measurable disease, no prior cytotoxic therapy, an ECOG performance status of 0 to 2, and evidence of disease progression while on progestins or after radiation therapy. Topotecan was administered at 1.5 mg/m(2) (or 1.2 mg/m(2) for patients with prior pelvic radiation) intravenously daily for 5 days every 3 weeks. RESULTS: A total of 44 patients were enrolled; 42 were eligible. The study was suspended because of unexpected toxicities, primarily sepsis and bleeding. After toxicity review, the study was reopened using lower doses of topotecan (1.0 mg/m(2) or 0.8 mg/m(2) for patients with prior radiation therapy). In addition, prophylactic use of growth factors was allowed after the first cycle, and patients with performance status of 2 were excluded. The major toxicities were hematologic and gastrointestinal. Among the 40 assessable patients, there were three (7.5%) complete responders and five partial responders (12.5%), for an overall response rate of 20%. The median duration of response was 8.0 months and of overall survival was 6.5 months. CONCLUSION: Topotecan is an active agent for the treatment of advanced endometrial carcinoma. At the doses and schedules initially used, toxicities were unacceptable; however, at the modified doses, toxicities were acceptable and clinical activity was preserved. PMID- 12775737 TI - Long-term follow-up of a prospective study of combined modality therapy for stage I-II indolent non-Hodgkin's lymphoma. AB - PURPOSE: Standard therapy for patients with stage I-II indolent lymphoma has been involved-field radiation therapy (IF-XRT), which achieves 10-year disease-free survival in 40% to 50% of patients, with many of these patients cured. We investigated the potential for combined-modality therapy to increase the disease free survival for such patients. PATIENTS AND METHODS: A total of 102 eligible patients with stage I-II low grade lymphoma (International Working Formulation criteria) were enrolled from 1984 to 1992. Treatment comprised 10 cycles of risk adapted chemotherapy (cyclophosphamide, vincristine, prednisone, bleomycin [COP Bleo], and with doxorubicin added for some [CHOP-Bleo]) and 30 to 40 Gy IF-XRT. RESULTS: The patients' median age was 56 years (range, 28 to 77), with follicular histology in 83%, bulky disease (>/= 5 cm) in 24%, and stage II in 52%. There were no treatment-related deaths and 99% of patients attained complete remission. With a median follow-up of 10 years, the 10-year time to treatment failure and overall survival were 76% and 82%, respectively. For patients with follicular lymphoma, these figures were 72% and 80%, respectively. The only factor associated with treatment failure, for follicular lymphoma patients, was stage modified International Prognostic Factors Index score (P =.02). None of 17 patients with diffuse small lymphocytic or mucosa-associated lymphoid tissue histology have relapsed. Elevated serum beta2-microglobulin was associated with shorter survival (P <.0001). The 10-year survival after relapse was 46%. There have been two cases of myelodysplasia and 12 other new malignancies, including four arising within radiation fields. CONCLUSION: With prolonged follow-up, combined-modality therapy with risk-adapted COP-/CHOP-Bleo and IF radiation has attained higher rates of disease control and survival than previously reported with IF-XRT alone. This apparent improvement is being further explored in an ongoing randomized trial. PMID- 12775738 TI - Therapy-related acute promyelocytic leukemia. AB - PURPOSE: To analyze patient cases of therapy-related acute promyelocytic leukemia (tAPL), occurring after chemotherapy (CT), radiotherapy (RT) or both for a prior disorder, diagnosed during the last 20 years in three European countries. PATIENTS AND METHODS: The primary disorder and its treatment, interval from primary disorder to tAPL, characteristics of tAPL, and its outcome were analyzed in 106 patients. RESULTS: Eighty of the 106 cases of tAPL were diagnosed during the last 10 years, indicating an increasing incidence of tAPL. Primary disorders were predominantly breast carcinoma (60 patients), non-Hodgkin's lymphoma (15 patients), and other solid tumors (25 patients). Thirty patients had received CT alone, 27 patients had received RT alone, and 49 patients had received both. CT included at least one alkylating agent in 68 patients and at least one topoisomerase II inhibitor in 61 patients, including anthracyclines (30 patients), mitoxantrone (28 patients), and epipodophyllotoxins (19 patients). Median interval from primary disorder to tAPL diagnosis was 25 months (range, 4 to 276 months). Characteristics of tAPL were generally similar to those of de novo APL. With treatment using anthracycline-cytarabine-based CT or all-trans retinoic acid combined with CT, actuarial survival was 59% at 8 years. CONCLUSION: tAPL is not exceptional, and develops usually less than 3 years after a primary neoplasm (especially breast carcinoma) treated in particular with topoisomerase II-targeted drugs (anthracyclines or mitoxantrone and less often etoposide). Characteristics and outcome of tAPL seem similar to those of de novo APL. PMID- 12775739 TI - Quality of life in patients with newly diagnosed chronic phase chronic myeloid leukemia on imatinib versus interferon alfa plus low-dose cytarabine: results from the IRIS Study. AB - PURPOSE: Quality of life (QOL) outcomes in patients with chronic myeloid leukemia (CML) were evaluated in an international phase III study. PATIENTS AND METHODS: Newly diagnosed patients with chronic phase CML were randomly assigned to imatinib or interferon alfa plus subcutaneous low-dose cytarabine (IFN+LDAC). Cross-over to the other treatment was permitted because of intolerance or lack of efficacy. Patients completed cancer-specific QOL (Functional Assessment of Cancer Therapy-Biologic Response Modifiers) and utility (Euro QoL-5D) questionnaires at baseline and during treatment (n = 1,049). The primary QOL end point was the Trial Outcome Index (TOI; a measure of physical function and well-being). Secondary end points included social and family well-being (SFWB), emotional well being (EWB), and the utility score. Primary analyses were intention to treat with secondary analyses accounting for cross-over. RESULTS: Patients receiving IFN+LDAC experienced a large decline in the TOI, whereas those receiving imatinib maintained their baseline level. Treatment differences at each visit were significant (P <.001) and clinically relevant in favor of imatinib. Mean SFWB, EWB, and utility scores were also significantly better for those patients taking imatinib. Patients who crossed over to imatinib experienced a large increase in TOI; significant (P <.001) differences were observed between patients who did and did not cross over in favor of imatinib. CONCLUSION: Imatinib offers clear QOL advantages compared with IFN+LDAC as first-line treatment of chronic phase CML. In addition, patients who cross over to imatinib from IFN+LDAC experience a significant improvement in QOL compared with patients who continue to take IFN+LDAC. PMID- 12775740 TI - Variations in the 5-hydroxytryptamine type 3B receptor gene as predictors of the efficacy of antiemetic treatment in cancer patients. AB - PURPOSE: Serotonin (5-hydroxytryptamine type 3 [5-HT3]) receptor antagonists have substantially reduced but not eliminated nausea and vomiting in patients undergoing cancer chemotherapy. They act through specific binding to the 5-HT3A, 5-HT3B receptor complex. The 5-HT3B subunit seems to be most important for its functionality. We hypothesized that patients with genetic variations in the 5 HT3B receptor gene might respond differently to antiemetic treatment. PATIENTS AND METHODS: We included 242 cancer patients on their first day of chemotherapy. Nausea and vomiting were documented before and twice during the chemotherapy using standardized interviews and visual analog scales. We sequenced the entire 5 HT3B receptor gene, including the 5' flanking region and at least a 20-base pair intronic sequence of each intron-exon splice site of all patients. RESULTS: Approximately 30% of all patients suffered from nausea or vomiting. Sequencing of the 5-HT3B receptor gene revealed 13 polymorphisms: two of them were amino acid exchanges (Tyr129Ser, Ala223Thr) and two were deletion variants. In both observation periods, patients homozygous for the -100_-102delAAG deletion variant of the promotor region experienced vomiting more frequently than did all the other patients. CONCLUSION: A more efficient antiemetic treatment with 5-HT3 receptor antagonists might be possible on a pharmacogenetic basis. However, only a small fraction of the therapeutic failure is explained by the -AAG deletion variant of the 5-HT3B receptor gene. Additional clinical and biochemical studies are needed to confirm the association. PMID- 12775741 TI - Patient preference for radiotherapy fractionation schedule in the palliation of painful bone metastases. AB - PURPOSE: The radiotherapeutic management of painful bone metastases is controversial, with several institutional and national guidelines advocating use of single-fraction radiotherapy. We aimed to determine patient choice of fractionation schedule after involvement in the decision-making process by use of a decision board. PATIENTS AND METHODS: Advantages and disadvantages of two fractionation schedules (24 Gy in six fractions v 8 Gy in one fraction) used in the randomized Dutch Bone Metastasis Study were discussed with patients using a decision board. Patients were asked to choose a fractionation schedule, to give reasons for their choice, and to indicate level of satisfaction with being involved in decision making. RESULTS: Sixty-two patients were entered. Eighty five percent (95% confidence interval, 74% to 93%) chose 24 Gy in six fractions over 8 Gy in one fraction (P <.0005). Variables including age, sex, performance status, tumor type, pain score, and paying class were not significantly related to patient choice. Multiple fractionation was chosen for lower re-treatment rates (92%) and fewer fractures (32%). Single-fraction treatment was chosen for cost (11%) and convenience (89%). Eighty-four percent of patients expressed positive opinions about being involved in the decision-making process. CONCLUSION: Decision board instruments are feasible and acceptable in an Asian population. The vast majority of patients preferred 24 Gy fractionated radiotherapy compared with a single fraction of 8 Gy. These results indicate the need for further research in this important area and serve to remind both clinicians and national or institutional policy makers of the importance of individual patient preference in treatment decision making. PMID- 12775742 TI - Cancer-specific mortality after surgery or radiation for patients with clinically localized prostate cancer managed during the prostate-specific antigen era. AB - PURPOSE: To determine whether pretreatment risk groups shown to predict time to prostate cancer-specific mortality (PCSM) after treatment at a single institution retained that ability in a multi-institutional setting. PATIENTS AND METHODS: From 1988 to 2002, 7,316 patients treated in the United States at 44 institutions with either surgery (n = 4,946) or radiation (n = 2,370) for clinical stage T1c 2, N0 or NX, M0 prostate cancer made up the study cohort. A Cox regression analysis was performed to determine the ability of pretreatment risk groups to predict time to PCSM after treatment. The relative risk (RR) of PCSM and 95% confidence intervals (CIs) were calculated for the intermediate- and high-risk groups relative to the low-risk group. RESULTS: Estimates of non-PCSM 8 years after prostate-specific antigen (PSA) failure were 4% v 15% (surgery versus radiation; Plog rank =.002) compared with 13% v 18% (surgery versus radiation; Plog rank =.35) for patients whose age at the time of PSA failure was less than 70 as compared with >or= 70 years, respectively. The RR of PCSM after treatment for surgery-managed patients with high- or intermediate-risk disease was 14.2 (95% CI, 5.0 to 23.4; PCox <.0001) and 4.9 (95% CI, 1.7 to 8.1; PCox =.0037), respectively. These values were 14.3 (95% CI, 5.2 to 24.0; PCox <.0001) and 5.6 (95% CI, 2.0 to 9.3; PCox =.0012) for radiation-managed patients. CONCLUSION: This study provided evidence to support the prediction of time to PCSM after surgery or radiation on the basis of pretreatment risk groups for patients with clinically localized prostate cancer managed during the PSA era. PMID- 12775743 TI - Groups potentially at risk for making poorly informed decisions about entry into clinical trials for childhood cancer. AB - PURPOSE: Some patients may be at greater risk than others of enrolling on a randomized clinical trial (RCT) without fully understanding the implications. To investigate this possibility, this study poses the question, Do non-English speaking Latino parents of children with leukemia show differences in their discussion of and understanding of an RCT when compared with English-speaking minority parents and with English-speaking majority (ie, white) parents? This research hypothesizes that factors such as social and educational status and the role of language interpreters may account for significant differences in the discussion and understanding of an RCT among the three groups. PATIENTS AND METHODS: A total of 108 parents are reported on, all of whom were observed, interviewed, and audiotaped during informed-consent discussions about participation in an RCT with their child's oncologist. Comparisons among the groups were performed using chi2 tests and a one-way analysis of variance. RESULTS: Problems of consent-related communication and understanding were more frequent among parents of low social status who spoke little or no English than they were in the two other groups. Several factors may have contributed to this disparity, including language interpretation, social status, and prevailing cultural norms. CONCLUSION: Some patients may be at greater risk than others of enrolling on an RCT without fully understanding the implications of their decision to do so. Numerous factors may contribute to this disparity, including language interpretation, social status, and prevailing cultural norms. Some specific strategies are suggested to help address this disparity. PMID- 12775744 TI - Metastatic melanoma in pregnancy: risk of transplacental metastases in the infant. AB - PURPOSE: Although metastases to the fetus via the placenta are rare, melanoma is the most common culprit. When it occurs, maternally derived melanoma metastasis in the infant is almost invariably fatal. PATIENTS AND METHODS: This article reviews current guidelines for placental evaluation in pregnant women with metastatic melanoma and presents surveillance recommendations for their infants. Comprehensive literature reviews were performed on melanoma in pregnancy and melanoma metastasis to the placenta and fetus. The use of interferon alfa in the pediatric population was also reviewed. A comprehensive search of the MEDLINE database (1966 to 2002) was performed. Articles were reviewed and additional references were obtained from the bibliographies. Translation of non-English articles was performed, and authors of previous publications were contacted. RESULTS: Eighty-seven patients with placental or fetal metastasis were identified. Twenty-seven occurrences were attributed to melanoma (31%). The fetus was affected in six of 27 melanoma patients (22%), with five of six infants dying of disease. The use of high-dose interferon alfa adjuvant therapy in pediatric patients has not been reported. CONCLUSION: The placentas of women with known or suspected metastatic melanoma should be carefully examined grossly and histologically by pathologists. With placental involvement, fetal risk of melanoma metastasis is approximately 22%. Neonates delivered with concomitant placental involvement should be considered a high-risk population. The risk benefit ratio of adjuvant treatment for a potentially affected infant should be carefully weighed. PMID- 12775745 TI - High-dose chemotherapy with autologous stem-cell rescue in children and adults with newly diagnosed pineoblastomas. AB - PURPOSE: We evaluated the usefulness of a treatment regimen that included high dose chemotherapy (HDC) with autologous stem-cell rescue (ASCR) in patients with newly diagnosed pineoblastoma (PBL). PATIENTS AND METHODS: Twelve patients with PBL were initially treated with surgery and induction chemotherapy. All but two patients underwent radiotherapy. Subsequently, all patients received HDC using cyclophosphamide (CTX) + melphalan (MEL) or busulfan (Bu) + MEL regimens and ASCR. RESULTS: A total of six children and six adults with median ages of 4.2 (range, 0.3 to 19.8 years) and 23 years (range, 23 to 43.7 years), respectively, were treated according to this strategy. Four patients had metastatic disease confined to the neuraxis. Five of 12 patients (42%) had a complete tumor resection at diagnosis. Ten patients received radiotherapy at median doses of 36.0 and 59.4 Gy to the neuraxis and pineal region, respectively. Eleven patients received HDC with CTX + MEL, and one patient received BU + MEL followed by ASCR. Nine patients are alive with no evidence of disease recurrence at a median of 62 months from diagnosis (range, 28 to 125 months), including three patients with metastatic disease and two infants who did not receive any radiotherapy. Three patients have died of progressive disease at 19, 32, and 37 months from diagnosis, respectively. The actuarial 4-year progression-free and overall survivals are 69% (95% confidence interval [CI], 39% to 99%) and 71% (95% CI, 43% to 99%), respectively. CONCLUSION: The use of HDC in addition to radiotherapy seems to be an effective treatment for patients with newly diagnosed pineoblastoma. PMID- 12775747 TI - Critical review of complementary therapies for prostate cancer. AB - Despite its prominence as the most frequently diagnosed solid tumor among men in the United States, relatively little is known about the etiology of prostate cancer. Furthermore, research into treatment strategies for prostate cancer continues to lag behind research for the other most common cancers. At the same time, however, the popularity of complementary therapies among prostate cancer patients continues to grow. In this article, we provide a critical review of the most recent evidence for dietary modifications, food supplements, and herbs in prostate cancer prevention and treatment. Despite encouraging data for some of these interventions, even the strongest proponents of complementary therapy agree that only randomized controlled trials can provide sufficient evidence on which to create universal guidelines. However, such trials are highly complex and expensive, and they require lengthy follow-up. Until such trials are completed, an opportunity exists for health care professionals to improve their knowledge and understanding of the current evidence for or against complementary therapy in prostate cancer. PMID- 12775746 TI - Potential role of platelets in endothelial damage observed during treatment with cisplatin, gemcitabine, and the angiogenesis inhibitor SU5416. AB - PURPOSE: An increased incidence of thromboembolic events was observed during treatment with cisplatin-gemcitabine plus SU5416 (CG+SU5416), a tyrosine kinase inhibitor targeting the vascular endothelial growth factor (VEGF) receptor-1 and 2. Nine thromboembolic events occurred in eight of 19 patients. We performed an analysis of parameters of the coagulation cascade and vessel wall activation. MATERIALS AND METHODS: Markers for thrombin generation and endothelial cell activation were measured in three patients treated with CG+SU5416, two of whom developed a thromboembolic event. The results were compared with measurements in six patients treated with CG alone, and in 17 patients treated with SU5416 alone. RESULTS: During cycles 1 and 2 of treatment with CG+SU5416, a significant cycle dependent activation of both the coagulation cascade and endothelial cells occurred, whereas platelet counts decreased. Change in platelet number had a significant negative predictive effect on soluble (s)-E-selectin levels. Significant activation of the coagulation cascade only was observed in the patients treated with CG alone, whereas in patients treated with SU5416 alone, significant endothelial cell activation was observed. CONCLUSION: We hypothesize that endothelial cells deprived of VEGF after exposure to SU5416 became activated and more susceptible to damage during treatment with CG+SU5416, which was aggravated by a transient decrease in platelets, which are, among other things, carriers of VEGF. These results suggests that VEGF, in addition to being a permeability, proliferation, and migration factor, also is a maintenance and protection factor for endothelial cells, and that platelets may have a role in maintaining vascular integrity. PMID- 12775748 TI - Why not start with thalidomide? PMID- 12775749 TI - Unusual problems in breast cancer and a rare lung cancer case. Case 1. Clinical complete response of breast cancer metastases after trastuzumab-based immunotherapy. PMID- 12775750 TI - Unusual problems in breast cancer and a rare lung cancer case. Case 2. Aggressive fibromatosis of the chest wall arising near a breast prosthesis. PMID- 12775751 TI - Unusual problems in breast cancer and a rare lung cancer case. Case 4. Primary lymphoepithelioma-like carcinoma of the lung. PMID- 12775752 TI - Guidelines for the use of epoetin: have quality-of-life benefits been proven? PMID- 12775753 TI - Whether an increase in hemoglobin above 12 g/dL is of clinical benefit. PMID- 12775754 TI - Androgen augmentation of epoetin. PMID- 12775756 TI - Decoding neuronal spike trains: how important are correlations? AB - It has been known for >30 years that neuronal spike trains exhibit correlations, that is, the occurrence of a spike at one time is not independent of the occurrence of spikes at other times, both within spike trains from single neurons and across spike trains from multiple neurons. The presence of these correlations has led to the proposal that they might form a key element of the neural code. Specifically, they might act as an extra channel for information, carrying messages about events in the outside world that are not carried by other aspects of the spike trains, such as firing rate. Currently, there is no general consensus about whether this proposal applies to real spike trains in the nervous system. This is largely because it has been hard to separate information carried in correlations from that not carried in correlations. Here we propose a framework for performing this separation. Specifically, we derive an information theoretic cost function that measures how much harder it is to decode neuronal responses when correlations are ignored than when they are taken into account. This cost function can be readily applied to real neuronal data. PMID- 12775757 TI - Toward a detailed computational model for the mammalian circadian clock. AB - We present a computational model for the mammalian circadian clock based on the intertwined positive and negative regulatory loops involving the Per, Cry, Bmal1, Clock, and Rev-Erb alpha genes. In agreement with experimental observations, the model can give rise to sustained circadian oscillations in continuous darkness, characterized by an antiphase relationship between Per/Cry/Rev-Erbalpha and Bmal1 mRNAs. Sustained oscillations correspond to the rhythms autonomously generated by suprachiasmatic nuclei. For other parameter values, damped oscillations can also be obtained in the model. These oscillations, which transform into sustained oscillations when coupled to a periodic signal, correspond to rhythms produced by peripheral tissues. When incorporating the light-induced expression of the Per gene, the model accounts for entrainment of the oscillations by light-dark cycles. Simulations show that the phase of the oscillations can then vary by several hours with relatively minor changes in parameter values. Such a lability of the phase could account for physiological disorders related to circadian rhythms in humans, such as advanced or delayed sleep phase syndrome, whereas the lack of entrainment by light-dark cycles can be related to the non-24h sleep-wake syndrome. The model uncovers the possible existence of multiple sources of oscillatory behavior. Thus, in conditions where the indirect negative autoregulation of Per and Cry expression is inoperative, the model indicates the possibility that sustained oscillations might still arise from the negative autoregulation of Bmal1 expression. PMID- 12775758 TI - Descent of the larynx in chimpanzee infants. AB - The human larynx descends during infancy and the early juvenile periods, and this greatly contributes to the morphological foundations of speech development. This developmental phenomenon is believed to be unique to humans. This concept has formed a basis for paleoanthropological studies on the origin and evolution of human speech. We used magnetic resonance imaging to study the development of three living chimpanzees and found that their larynges also descend during infancy, as in human infants. This descent was completed primarily through the rapid descent of the laryngeal skeleton relative to the hyoid, but it was not accompanied by the descent of the hyoid itself. The descent is possibly associated with developmental changes of the swallowing mechanism. Moreover, it contributes physically to an increased independence between the processes of phonation and articulation for vocalization. Thus, the descent of the larynx and the morphological foundations for speech production must have evolved in part during hominoid evolution, and not in a single shift during hominid evolution. PMID- 12775759 TI - A postgenomic method for predicting essential genes at subsaturation levels of mutagenesis: application to Mycobacterium tuberculosis. AB - We describe a postgenomic in silico approach for identifying genes that are likely to be essential and estimate their proportion in haploid genomes. With the knowledge of all sites eligible for mutagenesis and an experimentally determined partial list of nonessential genes from genome mutagenesis, a Bayesian statistical method provides reasonable predictions of essential genes with a subsaturation level of random mutagenesis. For mutagenesis, a transposon such as Himar1 is suitable as it inserts randomly into TA sites. All of the possible insertion sites may be determined a priori from the genome sequence and with this information, data on experimentally hit TA sites may be used to predict the proportion of genes that cannot be mutated. As a model, we used the Mycobacterium tuberculosis genome. Using the Himar1 transposon, we created a genetically defined collection of 1,425 insertion mutants. Based on our Bayesian statistical analysis using Markov chain Monte Carlo and the observed frequencies of transposon insertions in all of the genes, we estimated that the M. tuberculosis genome contains 35% (95% confidence interval, 28%-41%) essential genes. This analysis further revealed seven functional groups with high probabilities of being enriched in essential genes. The PE-PGRS (Pro-Glu polymorphic GC-rich repetitive sequence) family of genes, which are unique to mycobacteria, the polyketide/nonribosomal peptide synthase family, and mycolic and fatty acid biosynthesis gene families were disproportionately enriched in essential genes. At subsaturation levels of mutagenesis with a random transposon such as Himar1, this approach permits a statistical prediction of both the proportion and identities of essential genes of sequenced genomes. PMID- 12775760 TI - UV wavelength-dependent regulation of transcription-coupled nucleotide excision repair in p53-deficient human cells. AB - Nucleotide excision repair (NER) prevents skin cancer by eliminating highly genotoxic cyclobutane pyrimidine dimers (CPDs) induced in DNA by the UVB component of sunlight. NER consists of two distinct but overlapping subpathways, i.e., global NER, which removes CPD from the genome overall, and transcription coupled NER (TCNER), which removes CPD uniquely from the transcribed strand of active genes. Previous investigations have clearly established that the p53 tumor suppressor plays a crucial role in the NER process. Here we used the ligation mediated PCR technique to demonstrate, at nucleotide resolution along two chromosomal genes in human cells, that the requirement for functional p53 in TCNER, but not in global NER, depends on incident UV wavelength. Indeed, relative to an isogenic p53 wild-type counterpart, p53-deficient human lymphoblastoid strains were shown to remove CPD significantly less efficiently along both the transcribed and nontranscribed strands of the c-jun and hprt loci after exposure to polychromatic UVB (290-320 nm). However, in contrast, after irradiation with 254-nm UV, p53 deficiency engendered less efficient CPD repair only along the nontranscribed strands of these target genes. The revelation of this intriguing wavelength-dependent phenomenon reconciles an apparent conflict between previous studies which used either UVB or 254-nm UV to claim, respectively, that p53 is required for, or plays no role whatsoever in, TCNER of CPD. Furthermore, our finding highlights a major caveat in experimental photobiology by providing a prominent example where the extensively used "nonsolar" model mutagen 254-nm UV does not accurately replicate the effects of environmentally relevant UVB. PMID- 12775761 TI - Predicting gene expression levels from codon biases in alpha-proteobacterial genomes. AB - Predicted highly expressed (PHX) genes in five currently available high G+C complete alpha-proteobacterial genomes are analyzed. These include: the nitrogen fixing plant symbionts Sinorhizobium meliloti (SINME) and Mesorhizobium loti (MESLO), the nonpathogenic aquatic bacterium Caulobacter crescentus (CAUCR), the plant pathogen Agrobacterium tumefaciens (AGRTU), and the mammalian pathogen Brucella melitensis (BRUME). Three of these genomes, SINME, AGRTU, and BRUME, contain multiple chromosomes or megaplasmids (>1 Mb length). PHX genes in these genomes are concentrated mainly in the major (largest) chromosome with few PHX genes found in the secondary chromosomes and megaplasmids. Tricarboxylic acid cycle and aerobic respiration genes are strongly PHX in all five genomes, whereas anaerobic pathways of glycolysis and fermentation are mostly not PHX. Only in MESLO (but not SINME) and BRUME are most glycolysis genes PHX. Many flagellar genes are PHX in MESLO and CAUCR, but mostly are not PHX in SINME and AGRTU. The nonmotile BRUME also carries many flagellar genes but these are generally not PHX and all but one are located in the second chromosome. CAUCR stands out among available prokaryotic genomes with 25 PHX TonB-dependent receptors. These are putatively involved in uptake of iron ions and other nonsoluble compounds. PMID- 12775762 TI - Protection and compensation in the influenza virus-specific CD8+ T cell response. AB - Influenza virus-specific CD8+ T cells generally recognize peptides derived from conserved, internal proteins that are not subject to antibody-mediated selection pressure. Prior exposure to any one influenza A virus (H1N1) can prime for a secondary CD8+ T cell response to a serologically different influenza A virus (H3N2). The protection afforded by this recall of established CD8+ T cell memory, although limited, is not negligible. Key characteristics of primary and secondary influenza-specific host responses are probed here with recombinant viruses expressing modified nucleoprotein (NP) and acid polymerase (PA) genes. Point mutations were introduced into the epitopes derived from the NP and PA such that they no longer bound the presenting H2Db MHC class I glycoprotein, and reassortant H1N1 and H3N2 viruses were made by reverse genetics. Conventional (C57BL/6J, H2b, and Ig+/+) and Ig-/- (muMT) mice were more susceptible to challenge with the single NP [HKx31 influenza A virus (HK)-NP] and PA (HK-PA) mutants, but unlike the Ig-/- mice, Ig+/+ mice were surprisingly resistant to the HK-NP/-PA double mutant. This virus was found to promote an enhanced IgG response resulting, perhaps, from the delayed elimination of antigen-presenting cells. Antigen persistence also could explain the increase in size of the minor KbPB1703 CD8+ T cell population in mice infected with the mutant viruses. The extent of such compensation was always partial, giving the impression that any virus specific CD8+ T cell response operates within constrained limits. It seems that the relationship between protective humoral and cellular immunity is neither simple nor readily predicted. PMID- 12775763 TI - Previously uncharacterized roles of platelet-activating factor acetylhydrolase 1b complex in mouse spermatogenesis. AB - Platelet-activating factor (PAF) has been shown to affect sperm motility and acrosomal function, thereby altering fertility. PAF acetylhydrolase 1b (PAFAH1B) hydrolyzes PAF and is composed of three subunits [the lissencephaly (LIS1) protein and alpha1 and alpha2 subunits] and structurally resembles a GTP hydrolyzing protein. Besides the brain, transcripts for Lis1, alpha1, and alpha2 are localized to meiotic and early haploid germ cells. Here, we report disruptions of the alpha2 (Pafah1b2) and alpha1 (Pafah1b3) genes in mice. Male mice homozygous null for alpha2(alpha2-/-) are infertile, and spermatogenesis is disrupted at mid- or late pachytene stages of meiosis or early spermiogenesis. Whereas mice homozygous mutant for alpha1(alpha1-/-) have normal fertility and normal spermatogenesis, those with disruptions of both alpha1 and alpha2 (alpha1 /-alpha2-/-) manifest an earlier disturbance of spermatogenesis with an onset at preleptotene or leptotene stages of meiosis. Testicular Lis1 protein levels are up-regulated in the alpha2-/- and alpha1-/-alpha2-/- mice. Lowering Lis1 levels by inactivating one allele of Lis1 in alpha2 null or alpha1/alpha 2 null genetic backgrounds (i.e., alpha2-/-Lis1+/- or alpha1-/-alpha2-/-Lis1+/- mice) restored spermatogenesis and male fertility. Our data provide evidence for unique roles of the PAFAH1B complex and, particularly, the lissencephaly protein Lis1 in spermatogenesis. PMID- 12775765 TI - Androgen receptor represses the neuroendocrine transdifferentiation process in prostate cancer cells. AB - Androgen-ablation therapy is an effective method for treating prostate cancer. However, prostate tumors that survive long-term androgen-ablation therapy are classified as androgen-independent as they proliferate in the absence of androgens, and they tend to be enriched for neuroendocrine (NE) cells. Androgen withdrawal causes androgen-dependent prostate cancer cells to adopt a pronounced NE phenotype, suggesting that androgen receptor (AR) represses an intrinsic NE transdifferentiation process in prostate cancer cells. In this report we show that short interfering RNA-induced AR silencing induced a NE phenotype that manifested itself in the growth of dendritic-like processes in both the androgen dependent LNCaP and androgen-independent LNCaP-AI human prostate cancer cells. Western blot analysis revealed that neuronal-specific enolase, a marker of the neuronal lineage, was increased by AR knockdown in LNCaP cells. The expression levels of the neuronal-specific cytoskeletal proteins beta-tubulin III, nestin, and glial acidic fibrillary protein were also characterized in AR knockdown cells. Most interestingly, AR silencing induced beta-tubulin III expression in LNCaP cells, while AR knockdown increased glial acidic fibrillary protein levels in both LNCaP and LNCaP-AI cells. Lastly, AR silencing reduced the proliferative capacity of LNCaP and LNCaP-AI cells. Our data demonstrate that AR actively represses an intrinsic NE transdifferentiation process in androgen-responsive prostate cancer cells and suggest a potential link between AR inactivation and the increased frequency of NE cells in androgen-independent tumors. PMID- 12775764 TI - Microarray analysis of uterine gene expression in mouse and human pregnancy. AB - Improved care of infants born prematurely has increased their survival. However, the incidence of preterm labor has not changed. To understand the processes involved in preterm labor, we used oligonucleotide microarrays to study gene expression in murine and human uterus during pregnancy. The induction of enzymes for prostaglandin synthesis was used as a marker for important changes during pregnancy because prostaglandins strongly contribute to both human and murine labor. We identified 504 genes that changed at least 2-fold between d 13.5 and 19.0 in the gravid mouse uterus. In the pregnant human myometrium, we found 478 genes that changed at least 2-fold in either term or preterm labor compared with preterm nonlabor specimens and 77 genes that significantly varied in both preterm and term labor. Patterns of gene regulation within functional groups comparing human preterm and term labor were similar, although the magnitude of change often varied. Surprisingly, few genes that changed significantly throughout pregnancy were the same in the mouse and human. These data suggest that functional progesterone withdrawal in human myometrium may not be the primary mechanism for labor induction, may implicate similar mechanisms for idiopathic preterm and term labor in humans, and may identify novel targets for further study. PMID- 12775767 TI - The different cardiac expression of the type 2 iodothyronine deiodinase gene between human and rat is related to the differential response of the Dio2 genes to Nkx-2.5 and GATA-4 transcription factors. AB - By producing T3 from T4, type 2 iodothyronine deiodinase (D2) catalyzes the first step in the cascade underlying the effect exerted by thyroid hormone. Type 2 iodothyronine deiodinase mRNA is expressed at high levels in human heart but is barely detectable in the corresponding rodent tissue. Although the heart is a major target of thyroid hormone, the role of cardiac D2 and the factors that regulate its expression are unknown. Here we report that the human Dio2 promoter is very sensitive to the cardiac transcription factors Nkx-2.5 and GATA-4. Nkx 2.5 transactivates a 6.5-kb human (h)Dio2-chloramphenicol acetyltransferase construct, with maximal induction reached with a 633-bp proximal promoter region. Interestingly, despite 73% identity with the corresponding human region, the rat Dio2 promoter is much less responsive to Nkx-2.5 induction. Using EMSA, we found that two sites in the human promoter (C and D) specifically bind Nkx-2.5. In coexpression studies, GATA-4 alone was a poor inducer of the hDio2 promoter; however in synergy with Nkx-2.5, it activated D2 reporter gene expression in the human, but not the rat promoter. Functional analysis showed that both C and D sites are required for the complete Nkx-2.5 response and for the Nkx-2.5/GATA-4 synergistic effect. In neonatal rat primary myocardiocytes, most of the hDio2 chloramphenicol acetyltransferase activity was suppressed by mutation of the Nkx 2.5 binding sites. Finally, a mutant Nkx-2.5 protein (N188K), which causes, in heterozygosity, congenital heart diseases, did not transactivate the Dio2 promoter and interfered with its activity in cardiomyocytes, possibly by titrating endogenous Nkx-2.5 protein away from the promoter. In conclusion, this study shows that Nkx-2.5 and GATA-4 play prime roles in Dio2 gene regulation in the human heart and suggests that it is their synergistic action in humans that causes the differential expression of the cardiac Dio2 gene between humans and rats. PMID- 12775766 TI - DAX-1, an unusual orphan receptor at the crossroads of steroidogenic function and sexual differentiation. AB - The unusual orphan member of the nuclear hormone receptor superfamily DAX-1 (NR0B1) owes its name to its double role in human pathology. On one side, duplications in Xp21, containing the DAX-1 gene, cause phenotypic sex reversal in XY individuals. On the other side, DAX-1 gene mutations are responsible for adrenal hypoplasia congenita, invariably associated with hypogonadotropic hypogonadism. DAX-1 functions as a global negative regulator of steroid hormone production by repressing the expression of multiple genes involved in the steroidogenic pathway. Here we review the mechanism of DAX-1 function in adrenal and gonadal differentiation, with special emphasis on recent results showing the critical role of DAX-1 protein misfolding in the pathogenesis of adrenal hypoplasia congenita. PMID- 12775768 TI - Mass spectrometric characterization of proteins from the SARS virus: a preliminary report. AB - A new coronavirus has been implicated as the causative agent of severe acute respiratory syndrome (SARS). We have used convalescent sera from several SARS patients to detect proteins in the culture supernatants from cells exposed to lavage another SARS patient. The most prominent protein in the supernatant was identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) as a approximately 46-kDa species. This was found to be a novel nucleocapsid protein that matched almost exactly one predicted by an open reading frame in the recently published nucleotide sequence of the same virus isolate (>96% coverage). A second viral protein corresponding to the predicted approximately 139-kDa spike glycoprotein has also been examined by MALDI-TOF MS (42% coverage). After peptide N-glycosidase F digestion, 12 glycosylation sites in this protein were confirmed. The sugars attached to four of the sites were also identified. These results suggest that the nucleocapsid protein is a major immunogen that may be useful for early diagnostics, and that the spike glycoprotein may present a particularly attractive target for prophylactic intervention in combating SARS. PMID- 12775769 TI - Breast cancer: when proteomics challenges biological complexity. AB - Proteomics is now entering into the field of biomedicine with declared hopes for the identification of new pathological markers and therapeutic targets. Current proteomic tools allow large-scale, high-throughput analyses for the detection, identification, and functional investigation of low-abundant proteins. However, the major limitation of proteomic investigations remains the complexity of biological structures and physiological processes, rendering the path of exploration of related pathologies paved with various difficulties and pitfalls. The case of breast cancer illustrates the major challenge facing modern proteomics and more generally post-genomics: to tackle the complexity of life. PMID- 12775771 TI - Getting the word out. By Caveman. PMID- 12775772 TI - Formins: signaling effectors for assembly and polarization of actin filaments. AB - Eukaryotic cells require filamentous actin to maintain their shape and for movement, growth and replication. New actin filaments are formed by the cutting of existing filaments or de novo through the action of specialized nucleators. The most highly characterized nucleator is the Arp2/3 complex, which nucleates the branched actin networks in the lamellae of migrating cells. Recently, Bni1p, which is a member of the formin family of proteins, has been shown to nucleate actin filaments in vitro. Formins are implicated in the formation of actin cables in yeast, stress fibers in tissue culture cells and cytokinesis in many cell types. Formins contain two highly conserved formin-homology domains, FH1 and FH2. The Bni1p FH2 domain is sufficient to mediate nucleation. The Bni1p FH1 domain binds profilin, an actin-monomer-binding protein that delivers actin to the growing barbed end of filaments. The Bni1p FH1-profilin interaction enhances nucleation. Formins participate in a number of signaling pathways that control the assembly of specific actin structures and bind the barbed end of actin filaments, thereby providing a cytoskeletal basis for the establishment of cell polarity. PMID- 12775773 TI - Regulation of F-actin-dependent processes by the Abl family of tyrosine kinases. AB - The F-actin cytoskeleton is a fundamental component of all eukaryotic cells. It provides force and stability and plays an integral role in a diverse array of cellular processes. The spatiotemporal regulation of F-actin dynamics is essential for proper biological output. The basic molecular machinery underlying the assembly and disassembly of filamentous actin is conserved in all eukaryotic cells. Additionally, protein tyrosine kinases, found only in multicellular eukaryotes, provide links between extracellular signals and F-actin-dependent cellular processes. Among the tyrosine kinases, c-Abl and its relative Arg are unique in binding directly to F-actin. Recent results have demonstrated a role for c-Abl in membrane ruffling, cell spreading, cell migration, and neurite extension in response to growth factor and extracellular matrix signals. c-Abl appears to regulate the assembly of F-actin polymers into different structures, depending on the extracellular signal. Interestingly, c-Abl contains nuclear import and export signals, and the nuclear c-Abl inhibits differentiation and promotes apoptosis in response to genotoxic stress. The modular structure and the nuclear-cytoplasmic shuttling of c-Abl suggest that it integrates multiple signals to coordinate F-actin dynamics with the cellular decision to differentiate or to die. PMID- 12775774 TI - Secreted antagonists of the Wnt signalling pathway. AB - The extracellular antagonists of the Wnt signalling pathway can be divided into two broad classes. Both classes of molecule prevent ligand-receptor interactions, but by different mechanisms: members of the first class, which include the sFRP (secreted Frizzled-related protein) family, WIF (Wnt inhibitory factor)-1 and Cerberus, primarily bind to Wnt proteins; the second class comprises certain members of the Dickkopf (Dkk) family, which bind to one subunit of the Wnt receptor complex. In addition, there are other protein interactions that contribute to Wnt antagonist function. Moreover, certain sFRPs and Dkks do not antagonise Wnt function, which suggests that these families have as-yet undiscovered functions. PMID- 12775778 TI - Iraq: time to focus our response. PMID- 12775780 TI - Age in epidemiological analysis. PMID- 12775779 TI - Towards legionnaires' disease control: epidemiological or environmental surveillance? PMID- 12775781 TI - Basic molecular genetics for epidemiologists. PMID- 12775782 TI - Arts for health: still searching for the Holy Grail. PMID- 12775783 TI - Can breast feeding modify the adverse effects of smoking during pregnancy on the child's cognitive development? PMID- 12775784 TI - Promotion of health and physical activity improves the mental health of elderly immigrants: results of a group randomised controlled trial among Turkish immigrants in the Netherlands aged 45 and over. AB - OBJECTIVES: Older immigrants from non-industrialised countries are a growing group, they have comparatively many health problems and are often hard to reach through health promotion and other preventive services. The aim of this study was to assess the effect of a short health education and physical exercise programme on the health and the physical activity of Turkish first generation elderly immigrants. DESIGN: Randomised controlled trial. SETTING: Welfare services in six Dutch cities. PARTICIPANTS: 126 people born in Turkey and aged 45 years and over, of whom 92 completed the trial. INTERVENTION: Eight, two hour sessions consisting of health education and exercises. Topics in health education focused on means to maintain a good health. Education was adapted to the culture and knowledge of older Turks and offered by a Turkish peer educator, in Turkish. MAIN OUTCOME MEASURES: Physical and mental wellbeing, and mental health based on the SF-12/36; knowledge on health and disease; physical activity. RESULTS: Participants were highly disadvantaged; 52% had not completed primary school and 49% had considerable problems in speaking Dutch. Participants in the intervention group showed an improvement in mental health (effect size: 0.38 SD (95% confidence intervals 0.03 to 0.73), p=0.03); the oldest subgroup also in mental wellbeing (effect size 0.75 SD (0.22 to 1.28), p=0.01). No improvements were seen in physical wellbeing and activity, nor in knowledge. CONCLUSIONS: Health education and physical exercise improve the mental state of deprived immigrants. Painstaking cultural adaptations to contents and method of delivery are essential to reach this effect. PMID- 12775785 TI - Smoking reduction, smoking cessation, and incidence of fatal and non-fatal myocardial infarction in Denmark 1976-1998: a pooled cohort study. AB - OBJECTIVE: To analyse the effects of smoking reduction and smoking cessation on incidence of myocardial infarction after adjustment for established cardiovascular risk factors. DESIGN: Prospective cohort study with record linkage to mortality and hospital registers. The association of individual change in smoking with myocardial infarction was examined in Cox proportional hazard analyses with continuous heavy smokers (> or =5 cigarettes/day) as reference. SETTING: Pooled data from three population studies conducted in Copenhagen, Denmark. PARTICIPANTS: 10 956 men and 8467 women with complete information on smoking habits at two examinations five to ten years apart were followed up from the second examination for a first hospital admission or death from myocardial infarction. Mean duration of follow up was 13.8 years. MAIN RESULTS: A total of 643 participants who were heavy smokers at baseline reduced their daily tobacco consumption by at least 50% without quitting between first and second examination, and 1379 participants stopped smoking. During follow up 1658 men and 521 women experienced a fatal or non-fatal myocardial infarction. After adjustment for cardiovascular risk factors, people who stopped smoking had a decreased risk of myocardial infarction, hazard ratio 0.71 (95% confidence intervals 0.59 to 0.85). Smoking reduction was not associated with reduced risk of myocardial infarction, hazard ratio 1.15 (95% confidence intervals 0.94 to 1.40). These associations remained unchanged after controlling for baseline illness in different ways. CONCLUSIONS: Smoking cessation in healthy people reduces the risk of a subsequent myocardial infarction, whereas this study provides no evidence of benefit from reduction in the amount smoked. PMID- 12775786 TI - Cervical screening and health inequality in England in the 1990s. AB - STUDY OBJECTIVE: s: To examine changing inequality in the coverage of cervical screening and its relation to organisational aspects of primary care and to inequality in cervical cancer incidence and mortality. DESIGN: Retrospective time trends analysis (1991-2001) of screening coverage and cervical cancer incidence and mortality in England. SETTING: The 99 district health authorities in England, as defined by 1999 boundaries were used to create a time series of incidence and mortality rates from cervical cancer per 100 000 population. A subset of 60 district health authorities were used to construct a time series of screening coverage data and GP and practice characteristics. Health authorities were categorised into one of three "deprivation" groups using the Townsend Deprivation Index. PARTICIPANTS: Women aged <35 and 35-64 were selected from health authority populations as the main focus of the study. RESULTS: Cervical cancer screening coverage was consistently higher in affluent areas from 1991-9 but ratio rates of inequality between affluent and deprived health authorities narrowed over time. The increase in coverage in deprived areas was most closely associated with an increase in the number of practice nurses. Cervical cancer incidence and mortality rates were consistently higher in deprived health authorities, but inequality decreased. Screening coverage and cervical cancer rates were highly negatively correlated in deprived health authorities. CONCLUSION: A primary health care intervention such as an organised programme of cervical screening can contribute to reducing inequality in population health. PMID- 12775787 TI - Socioeconomic disparities in health care use: Does universal coverage reduce inequalities in health? AB - BACKGROUND: Despite enormous public sector expenditures, the effectiveness of universal coverage for health care in reducing socioeconomic disparities in health has received little attention. STUDY OBJECTIVE: s: To evaluate whether universal coverage for health care reduces socioeconomic disparities in health. DESIGN: Information on participants of the 1990 Nova Scotia Nutrition Survey was linked with eight years of administrative health services data and mortality. The authors first examined whether lower socioeconomic groups use more health services, as would be expected given their poorer health status. They then investigated to what extent differential use of health services modifies socioeconomic disparities in mortality. Finally, the authors evaluated health services use in the last years of life when health is poor regardless of a person's socioeconomic background. SETTING: The Canadian province of Nova Scotia, which provides universal health care coverage to all residents. PARTICIPANTS: 1816 non-institutionalised adults, aged 18-75 years, from a two stage cluster sample stratified by age, gender, and region. MAIN RESULTS: People with lower socioeconomic background used comparatively more family physician and hospital services, in such a way as to ameliorate the socioeconomic differences in mortality. In contrast, specialist services were comparatively underused by people in lower socioeconomic groups. In the last three years of life, use of specialist services was significantly higher in the highest income group. CONCLUSIONS: Universal coverage of family physician and hospital services ameliorate the socioeconomic differences in mortality. However, specialist services are underused in lower socioeconomic groups, bearing the potential to widen the socioeconomic gap in health. PMID- 12775788 TI - A matter of life and death: population mortality and football results. AB - OBJECTIVES: To determine whether football results are associated with mortality from circulatory disease. DESIGN: Retrospective study, comparing mortality on days of football matches between 18 August 1994 and 28 December 1999 with the results of the football matches. SETTING: Newcastle and North Tyneside, Sunderland, Tees, and Leeds Health Authority areas of England. SUBJECTS: All persons resident in Newcastle and North Tyneside, Sunderland, Tees, and Leeds Health Authority areas of England. MAIN OUTCOME MEASURES: Mortality attributable to acute myocardial infarction and stroke. RESULTS: On days when the local professional football team lost at home, mortality attributable to acute myocardial infarction and stroke increased significantly in men (relative risk 1.28, 95% confidence intervals 1.11 to 1.47). No increase was observed in women. CONCLUSIONS: Results achieved by the local professional football team are associated systematically with circulatory disease death rates over a five year period in men, but not women. PMID- 12775790 TI - Register based monitoring shows decreasing socioeconomic differences in Finnish perinatal health. AB - STUDY OBJECTIVE: Several studies on differences in infant outcome by socioeconomic position have been done, but these have usually been based on ad hoc data linkages. The aim of this paper was to investigate whether socioeconomic differences in perinatal health in Finland could be regularly monitored using routinely collected data from one single register. DESIGN AND SETTING: Since October 1990, the Finnish Medical Birth Register (MBR) has included data on maternal occupation. A special computer program that converted the occupation name into an occupational code and into a socioeconomic position was prepared. Perinatal health was measured with five different indicators. The Finnish MBR data for years 1991 to 1999 (n=565 863 newborns) were used in the study. The study period was divided into three, three year periods to study time trends. RESULTS: An occupational code was derived for 95% of women, but it was not possible to define a socioeconomic position for 22% of women, including, for example, students and housewives (the group "Others"). For the rest, the data showed socioeconomic differences in all perinatal health indicators. Maternal smoking explained up to half of the excess risk for adverse perinatal outcome in the lowest socioeconomic group. The socioeconomic differences narrowed during the 1990s: infant outcome improved in the lowest socioeconomic group, but remained at the same level or even deteriorated in other groups. When comparing the lowest group with the highest group, the odds ratios (OR) adjusted for maternal background characteristics at least halved for prematurity (from 1.32 (95% confidence intervals 1.24 to 1.43) in 1991-1993 to 1.16 (1.08 to 1.25) in 1997 1999), for low birth weight (from 1.49 (1.36 to 1.63) to 1.25 (1.17 to 1.40)), and for perinatal mortality (from 1.79 (1.44 to 2.21) to 1.33 (1.07 to 1.66)). CONCLUSIONS: Social inequality in perinatal health outcomes exists in Finland, but seems to have diminished in the 1990s. These data showed that routinely collected birth register data provide a good source for studies on socioeconomic health differences in the perinatal period, but that uncertainty, mainly attributable to the large group of women with difficult to classify socioeconomic status, remains. PMID- 12775791 TI - Socioeconomic differences in attitudes and beliefs about healthy lifestyles. AB - STUDY OBJECTIVE: s: The factors underlying socioeconomic status differences in smoking, leisure time physical activity, and dietary choice are poorly understood. This study investigated attitudes and beliefs that might underlie behavioural choices, including health locus of control, future salience, subjective life expectancy, and health consciousness, in a nationally representative sample. DESIGN: Data were collected as part of the monthly Omnibus survey of the Office of National Statistics in Britain. PARTICIPANTS: A stratified, probability sample of 2728 households was selected by random sampling of addresses. One adult from each household was interviewed. MAIN RESULTS: Higher SES respondents were less likely to smoke and more likely to exercise and eat fruit and vegetables daily. Lower SES was associated with less health consciousness (thinking about things to do to keep healthy), stronger beliefs in the influence of chance on health, less thinking about the future, and lower life expectancies. These attitudinal factors were in turn associated with unhealthy behavioural choices, independently of age, sex, and self rated health. CONCLUSIONS: Socioeconomic differences in healthy lifestyles are associated with differences in attitudes to health that may themselves arise through variations in life opportunities and exposure to material hardship and ill health over the life course. PMID- 12775792 TI - Influence of individual and neighbourhood socioeconomic status on mortality among black, Mexican-American, and white women and men in the United States. AB - STUDY OBJECTIVE: s: This study examines the influence of individual and neighbourhood socioeconomic status (SES) on mortality among black, Mexican American, and white women and men in the US. The authors had three study objectives. Firstly, they examined mortality rates by both individual level SES (measured by income, education, and occupational/employment status) and neighbourhood level SES (index of neighbourhood income/wealth, educational attainment, occupational status, and employment status). Secondly, they examined whether neighbourhood SES was associated with mortality after controlling for individual SES. Thirdly, they calculated the population attributable risk to estimate the reduction in mortality rates if all women and men lived in the highest SES neighbourhoods. DESIGN: National Health Interview Survey (1987-1994), linked with 1990 census tract (neighbourhood proxy) and mortality data through 1997. SETTING/PARTICIPANTS: Nationally representative sample of 59 935 black, 19 201 Mexican-American, and 344 432 white men and women (six gender and racial/ethnic groups), aged 25-64 at interview. MAIN RESULTS: Mortality rates for all six gender and racial/ethnic groups were two to four times higher for those with the lowest incomes (lowest quartile) who lived in the lowest SES neighbourhoods (lowest tertile) compared with those with the highest incomes who lived in the highest SES neighbourhoods. For the six groups, the age adjusted mortality risk associated with living in the lowest SES neighbourhoods ranged from 1.43 to 1.61. The mortality risk decreased but remained significant (p values <.05) after adjusting for each of the three individual measures of SES, with the exception of Mexican-American women. Furthermore, the mortality risk associated with living in the lowest SES neighbourhoods remained significant after simultaneously adjusting for all three individual measures of SES for white men (p<0.001) and white women (p<0.05). Deaths would hypothetically be reduced by about 20% for each subgroup if everyone had the same death rates as those living in the highest SES neighbourhoods (highest tertile). CONCLUSIONS: Living in a low SES neighbourhood confers additional mortality risk beyond individual SES. PMID- 12775793 TI - Socioeconomic position, occupational exposures, and gender: the relation with locomotor disability in early old age. PMID- 12775795 TI - Adverse pregnancy outcomes around incinerators and crematoriums in Cumbria, north west England, 1956-93. AB - STUDY OBJECTIVE: To investigate the risk of stillbirth, neonatal death, and lethal congenital anomaly among babies of mothers living close to incinerators and crematoriums in Cumbria, north west England, 1956-93. DESIGN: Retrospective cohort study. Logistic regression was used to investigate the risk of each outcome in relation to proximity at birth to incinerators and crematoriums, adjusting for social class, year of birth, birth order, and multiple births. Continuous odds ratios for trend with proximity to sites were estimated. SETTING: All 3234 stillbirths, 2663 neonatal deaths, and 1569 lethal congenital anomalies among the 244 758 births to mothers living in Cumbria, 1956-1993. MAIN RESULTS: After adjustment for social class, year of birth, birth order, and multiple births, there was an increased risk of lethal congenital anomaly, in particular spina bifida (odds ratio 1.17, 95% CI: 1.07 to 1.28) and heart defects (odds ratio 1.12, 95% CI: 1.03 to 1.22) around incinerators and an increased risk of stillbirth (odds ratio 1.04, 95% CI: 1.01 to 1.07) and anencephalus (odds ratio 1.05, 95% CI: 1.00 to 1.10) around crematoriums. CONCLUSIONS: The authors cannot infer a causal effect from the statistical associations reported in this study. However, as there are few published studies with which to compare our results, the risk of spina bifida, heart defects, stillbirth, and anencephalus in relation to proximity to incinerators and crematoriums should be investigated further, in particular because of the increased use of incineration as a method of waste disposal. PMID- 12775796 TI - Association between offspring birth weight and atherosclerosis in middle aged men and women: British Regional Heart Study. PMID- 12775797 TI - Childhood mental ability and smoking cessation in adulthood: prospective observational study linking the Scottish Mental Survey 1932 and the Midspan studies. PMID- 12775799 TI - Using ACME (Automatic Classification of Medical Entry) software to monitor and improve the quality of cause of death statistics. PMID- 12775798 TI - Sporadic cases of community acquired legionnaires' disease: an ecological study to identify new sources of contamination. AB - STUDY OBJECTIVE: To explore the relation between incidence of sporadic and community acquired legionnaires' disease and exposure to potentially contaminated industrial aerosols. DESIGN: Geographical ecological approach using the postcode as the statistical unit. A multivariate Poisson regression model was used to model the relation between exposure to industrial aerosols and legionnaires' disease. SETTING: Metropolitan France. MAIN RESULTS: More than 1000 sources of industrial exposure (aerosol and plume of smoke) were identified in 42 French departments. After adjusting for confounding factors, there was a statistically increased incidence of legionnaires' disease in postcodes with plume of smoke in comparison with postcodes without (RR=1.45, 95% CI=1.12 to 1.87), and in postcodes with more than one aerosol in comparison with postcodes without (RR=1.37, 95% CI=1.04 to 1.79). CONCLUSION: These findings highlight that any industrial systems generating water aerosols should be regarded as potential sources of contamination for legionnaires' disease. PMID- 12775800 TI - Seasonality of live birth sex ratio in south western Siberia, Russia, 1959-2001. PMID- 12775801 TI - Validity and ethics in science. PMID- 12775802 TI - Data access. Industry groups petition for data on salt and hypertension. PMID- 12775803 TI - Infectious diseases. Clues to the animal origins of SARS. PMID- 12775804 TI - Environmental Protection Agency. Whitman leaves science legacy at EPA. PMID- 12775805 TI - National Science Foundation. Secrecy on big projects breeds earmarks, panel is told. PMID- 12775806 TI - Scientific workforce. Report asks colleges to plug a leaky people pipeline. PMID- 12775808 TI - Italian science. Industrial renaissance or a new dark age? PMID- 12775807 TI - Genetics. First cloned mule races to finish line. PMID- 12775810 TI - Astronomy. The Milky Way's dark, starving pit. PMID- 12775809 TI - Conservation biology. Experts say big cats don't leave useful tracks. PMID- 12775811 TI - Nuclear proliferation. North Korea's not-so-hidden agenda raises the ante. PMID- 12775812 TI - Nuclear proliferation. Pick your poison: U vs. Pu. PMID- 12775813 TI - Military research. U.S. defense labs brace for a blast from their bosses. PMID- 12775814 TI - Infectious diseases. Taming pathogens: an elegant idea, but does it work? PMID- 12775815 TI - Infectious diseases. Darwinian vaccines. PMID- 12775816 TI - Marine biology. Will oil spell trouble for Western Pacific gray whales? PMID- 12775817 TI - Fernando Reinach profile. From bench to boardroom: promoting Brazilian biotech. PMID- 12775818 TI - Retraction. PMID- 12775819 TI - Incorporating science into decision-making. PMID- 12775821 TI - Giving credit to the first linear collider. PMID- 12775820 TI - Evolution of protein amino acids. PMID- 12775822 TI - Comment on "A new species of yunnanozoan with implications for deuterostome evolution". PMID- 12775824 TI - Calculating the benefits of regulation. PMID- 12775825 TI - Patent law. Natural substances and patentable inventions. PMID- 12775826 TI - Virology. The SARS coronavirus: a postgenomic era. PMID- 12775828 TI - Geology. The Rodinia jigsaw puzzle. PMID- 12775827 TI - Materials science. Watching nanoparticles grow. PMID- 12775829 TI - Physics. Broken Cooper pairs caught bouncing around. PMID- 12775830 TI - Microbiology and evolution. Modulating mutation rates in the wild. PMID- 12775831 TI - Electron transport in molecular wire junctions. AB - Molecular conductance junctions are structures in which single molecules or small groups of molecules conduct electrical current between two electrodes. In such junctions, the connection between the molecule and the electrodes greatly affects the current-voltage characteristics. Despite several experimental and theoretical advances, including the understanding of simple systems, there is still limited correspondence between experimental and theoretical studies of these systems. PMID- 12775832 TI - Eukaryotic intron loss. PMID- 12775833 TI - Stress-induced mutagenesis in bacteria. AB - The evolutionary significance of stress-induced mutagenesis was evaluated by studying mutagenesis in aging colonies (MAC) of Escherichia coli natural isolates. A large fraction of isolates exhibited a strong MAC, and the high MAC variability reflected the diversity of selective pressures in ecological niches. MAC depends on starvation, oxygen, and RpoS and adenosine 3',5'-monophosphate regulons; thus it may be a by-product of genetic strategies for improving survival under stress. MAC could also be selected through beneficial mutations that it generates, as shown by computer modeling and the patterns of stress inducible and constitutive mutagenesis. We suggest that irrespective of the causes of their emergence, stress-induced mutations participate in adaptive evolution. PMID- 12775834 TI - Diffusion of nonequilibrium quasi-particles in a cuprate superconductor. AB - We report a transport study of nonequilibrium quasi-particles in a high transition-temperature cuprate superconductor using the transient grating technique. Low-intensity laser excitation (at a photon energy of 1.5 electron volts) was used to introduce a spatially periodic density of quasi-particles into a high-quality untwinned single crystal of YBa2Cu3O6.5. Probing the evolution of the initial density through space and time yielded the quasi-particle diffusion coefficient and the inelastic and elastic scattering rates. The technique reported here is potentially applicable to precision measurements of quasi particle dynamics not only in cuprate superconductors but in other electronic systems as well. PMID- 12775835 TI - A bond-fluctuation mechanism for stochastic switching in wired molecules. AB - Stochastic on-off conductivity switching observed in phenylene-ethynylene oligomers has been explained in terms of changes in ring conformations, or electron localization, or both. We report the observation of stochastic on-off switching in the simplest of wired molecules: octanedithiol, decanedithiol, and dodecanedithiol bonded on an Au(111) surface. Stochastic switching was observed even when a top gold contact was pressed on by a conducting atomic force microscope tip at constant force. The rate of switching increased substantially at 60 degrees C, a temperature at which these films are commonly annealed. Because such switching in alkanethiols is unlikely to be caused by internal molecular electronic changes and cannot be fully accounted for by breaking of the top contact, we argue that the cause is the well-known mobility of molecules tethered to gold via a thiol linkage. PMID- 12775837 TI - Atomic resolution imaging of a carbon nanotube from diffraction intensities. AB - Atomic imaging of three-dimensional structures has required a crystal in diffraction or a lens in electron imaging. Whereas diffraction achieves very high resolution by averaging over many cells, imaging gives localized structural information, such as the position of a single dopant atom. However, lens aberrations limit electron imaging resolution to about 1 angstrom. Resolution is reduced further by low contrast from weakscattering or from the limitations on electron dose for radiation-sensitive molecules. We show that both high resolution and high contrast can be achieved by imaging from diffraction with a nanometer-sized coherent electron beam. The phase problem is solved by oversampling and iterative phase retrieval. We apply this technique to image a double-wall carbon nanotube at 1-angstrom resolution, revealing the structure of two tubes of different helicities. Because the only requirement for imaging is a diffraction pattern sampled below the Nyquist frequency, our technique has the potential to image nonperiodic nanostructures, including biological macromolecules, at diffraction intensity-limited resolutions. PMID- 12775836 TI - Real-time monitoring of growing nanoparticles. AB - One challenge in the production of nanometer-sized objects with given properties is to control their growth at a macroscopic scale in situ and in real time. A dedicated ultrahigh-vacuum grazing-incidence small-angle x-ray scattering setup has been developed, yielding high sensitivity and dynamics. Its capabilities to derive the average particle shape and size and the film growth mode and ordering and to probe both surfaces and buried interfaces are illustrated for two prototypical cases: the model catalyst Pd/MgO(100) and the self-organized Co/Au(111) system. A wide range of technologically important systems can potentially be investigated in various gaseous environments. PMID- 12775838 TI - Seismic imaging of the downwelling Indian lithosphere beneath central Tibet. AB - A tomographic image of the upper mantle beneath central Tibet from INDEPTH data has revealed a subvertical high-velocity zone from approximately 100- to approximately 400-kilometers depth, located approximately south of the Bangong Nujiang Suture. We interpret this zone to be downwelling Indian mantle lithosphere. This additional lithosphere would account for the total amount of shortening in the Himalayas and Tibet. A consequence of this downwelling would be a deficit of asthenosphere, which should be balanced by an upwelling counterflow, and thus could explain the presence of warm mantle beneath north-central Tibet. PMID- 12775839 TI - Modulation of phospholipid signaling by GLABRA2 in root-hair pattern formation. AB - The root-hair pattern of Arabidopsis is determined through a regulatory circuit composed of transcription factor genes. The homeobox gene GLABRA2 (GL2) has been considered a key component, acting farthest downstream in this regulation. GL2 modified to include a transactivating function caused epidermal cells to develop ectopic root hairs or root hair-like structures. With this system, the phospholipase Dzeta1 gene (AtPLDzeta1) was identified as a direct target of GL2. Inducible expression of AtPLDzeta1 promoted ectopic root-hair initiation. We conclude that GL2 exerts its regulatory effect on root-hair development through modulation of phospholipid signaling. PMID- 12775840 TI - Disruption of the epithelial apical-junctional complex by Helicobacter pylori CagA. AB - Helicobacter pylori translocates the protein CagA into gastric epithelial cells and has been linked to peptic ulcer disease and gastric carcinoma. We show that injected CagA associates with the epithelial tight-junction scaffolding protein ZO-1 and the transmembrane protein junctional adhesion molecule, causing an ectopic assembly of tight-junction components at sites of bacterial attachment, and altering the composition and function of the apical-junctional complex. Long term CagA delivery to polarized epithelia caused a disruption of the epithelial barrier function and dysplastic alterations in epithelial cell morphology. CagA appears to target H. pylori to host cell intercellular junctions and to disrupt junction-mediated functions. PMID- 12775841 TI - Water-soluble quantum dots for multiphoton fluorescence imaging in vivo. AB - The use of semiconductor nanocrystals (quantum dots) as fluorescent labels for multiphoton microscopy enables multicolor imaging in demanding biological environments such as living tissue. We characterized water-soluble cadmium selenide-zinc sulfide quantum dots for multiphoton imaging in live animals. These fluorescent probes have two-photon action cross sections as high as 47,000 Goeppert-Mayer units, by far the largest of any label used in multiphoton microscopy. We visualized quantum dots dynamically through the skin of living mice, in capillaries hundreds of micrometers deep. We found no evidence of blinking (fluorescence intermittency) in solution on nanosecond to millisecond time scales. PMID- 12775842 TI - Disrupted timing of discontinuous but not continuous movements by cerebellar lesions. AB - Patients with cerebellar damage are known to exhibit deficits in the temporal control of movements. We report that these deficits are restricted to discontinuous movements. Cerebellar patients exhibited no deficit in temporal variability when producing continuous, rhythmic movements. We hypothesize that the temporal properties of continuous movements are emergent and reflect the operation of other control parameters not associated with the cerebellum. In contrast, discontinuous movements require an explicit representation of the temporal goal, a function of the cerebellum. The requirement for explicit temporal representation provides a parsimonious account of cerebellar involvement in a range of tasks. PMID- 12775843 TI - Characterization of mammalian selenoproteomes. AB - In the genetic code, UGA serves as a stop signal and a selenocysteine codon, but no computational methods for identifying its coding function are available. Consequently, most selenoprotein genes are misannotated. We identified selenoprotein genes in sequenced mammalian genomes by methods that rely on identification of selenocysteine insertion RNA structures, the coding potential of UGA codons, and the presence of cysteine-containing homologs. The human selenoproteome consists of 25 selenoproteins. PMID- 12775844 TI - Finding functional features in Saccharomyces genomes by phylogenetic footprinting. AB - The sifting and winnowing of DNA sequence that occur during evolution cause nonfunctional sequences to diverge, leaving phylogenetic footprints of functional sequence elements in comparisons of genome sequences. We searched for such footprints among the genome sequences of six Saccharomyces species and identified potentially functional sequences. Comparison of these sequences allowed us to revise the catalog of yeast genes and identify sequence motifs that may be targets of transcriptional regulatory proteins. Some of these conserved sequence motifs reside upstream of genes with similar functional annotations or similar expression patterns or those bound by the same transcription factor and are thus good candidates for functional regulatory sequences. PMID- 12775845 TI - Asteroseismology of HD 129929: core overshooting and nonrigid rotation. AB - We have gathered and analyzed 1493 high-quality multicolor Geneva photometric data taken over 21 years of the B3Vstar HD 129929. We detect six frequencies, among which appear the effects of rotational splitting with a spacing of approximately 0.0121 cycles per day, which implies that the star rotates very slowly. A nonadiabatic analysis of the oscillations allows us to constrain the metallicity of the star to Z epsilon [0.017,0.022], which agrees with a similar range derived from spectroscopic data. We provide evidence for the occurrence of core convective overshooting in the star, with alpha(ov) = 0.10 +/- 0.05, and we rule out rigid rotation. PMID- 12775846 TI - A mule cloned from fetal cells by nuclear transfer. PMID- 12775847 TI - Colorectal neoplasms: role of intravenous contrast-enhanced CT colonography. AB - PURPOSE: To evaluate whether computed tomographic (CT) colonography with intravenously administered contrast material can help predict malignant differentiation of colorectal neoplasms (> or =10 mm in diameter). MATERIALS AND METHODS: Enhancement of 29 consecutive colorectal neoplasms on pre- and postcontrast CT colonographic images was retrospectively measured. The neoplasms were subsequently resected. Enhancement was calculated by subtraction of attenuation values (in Hounsfield units) obtained with precontrast and postcontrast 45-second-delay prone CT colonographic sequences. The neoplasms were graded as follows: grade 1, adenoma; grade 2, adenoma with high-grade dysplasia; grade 3, well-differentiated adenocarcinoma; grade 4, moderately differentiated adenocarcinoma; and grade 5, poorly differentiated adenocarcinoma. Correlation among size, histologic grade, and degree of enhancement was made with Pearson and Spearman coefficients. The ability of the degree of enhancement to help predict adenocarcinoma (histologic grade, > or =3) was calculated. RESULTS: Histologic-CT colonographic correlation was performed in 29 neoplasms (mean diameter, 27.9 mm; range, 10-65 mm). There was no correlation between size and degree of enhancement, size and histologic grade (R = -0.17, P =.33), or histologic grade and degree of enhancement (R = 0.23, P =.23). However, increasing enhancement was noted between grades 2 and 5. When an enhancement threshold of 40 HU was used for the diagnosis of adenocarcinoma (grades 3-5), sensitivity was 92%, specificity was 20%, positive predictive value was 50%, and negative predictive value was 75%. CONCLUSION: The degree of contrast enhancement on a 45-second-delay CT colonographic image does not correlate with size or degree of histologic differentiation, although increasing enhancement with lesser degrees of differentiation was noted. PMID- 12775848 TI - Brain imaging findings in pediatric patients with sickle cell disease. AB - PURPOSE: To determine prevalence of imaging abnormalities in the brain of children with sickle cell disease (SCD) and to identify clinical and methodological factors that influence prevalence estimate. MATERIALS AND METHODS: Magnetic resonance (MR) imaging and MR angiographic findings for 185 patients with SCD examined at St Jude Children's Research Hospital since 1993 were reviewed. At least two readers independently reviewed images. Standard MR imaging criteria were used to identify lacunae, loss of white matter volume, encephalomalacia, or leukoencephalopathy. Patients were assigned grades to indicate limited or extensive abnormalities. Standard MR angiographic criteria were used to identify arterial tortuosity (limited vasculopathy) and stenosis or occlusion (extensive vasculopathy). Findings were evaluated as a function of patient clinical status (including stroke) and diagnosis. Recent methods (T1- and T2-weighted MR imaging plus fluid-attenuated inversion recovery [FLAIR] at 3-mm section thickness) were compared with older methods (T1- and T2-weighted MR imaging without FLAIR at 5-mm section thickness). RESULTS: At mean age of 10 years, overall prevalence of infarction, ischemia, or atrophy in patients with SCD was 44% (82 of 185), and prevalence of vasculopathy was 55% (102 of 185), without evidence of a significant referral bias. Twenty-six of 27 patients with clinical stroke had abnormal findings at imaging, but even if patients with stroke were excluded, 35% (56 of 158) had a "silent infarction" (MR imaging visible injury without clinical stroke), and 49% (78 of 158) had abnormal findings at MR angiography. Patients with clinically severe disease had more abnormalities at MR imaging (P <.001) and MR angiography (P <.004) than did patients with milder disease. Severe vasculopathy was more prevalent in patients with hemoglobin SS than in those with hemoglobin SC (P <.001). Recent imaging methods showed more abnormalities than did older methods (P <.01). With newer methods, 43% (29 of 67) of patients had extensive abnormalities, whereas with older methods, 28% (33 of 116) had extensive abnormalities. CONCLUSION: Prevalence of ischemic brain injury in pediatric patients with SCD is substantially higher than was previously reported, in part because of improvements in imaging methods. PMID- 12775849 TI - Obturator externus bursa: anatomic origin and MR imaging features of pathologic involvement. AB - PURPOSE: To investigate the anatomy of the obturator externus bursa of the hip and describe the magnetic resonance (MR) imaging features of pathologic involvement. MATERIALS AND METHODS: The authors dissected eight cadaver hemipelvises to assess for the presence of periarticular bursae of the hip and bursal communication with the fascial plane of the obturator externus muscle. In addition, 10 consecutive patients with obturator externus bursa enlargement were prospectively identified with MR imaging. A bursa was considered present when a fluid collection was seen extending along the obturator externus muscle, continuous with the posterior inferior hip joint. The direction, extent, contour, and thickness of the bursa and hip capsule were recorded. Surgical findings were available for eight of the 10 patients, with histopathologic correlation between the bursal lining and hip capsule. RESULTS: At cadaveric dissection, one specimen showed a bursa communicating with and extending from the posteroinferior aspect of the hip joint deep to the obturator externus tendon. In all 10 patients, MR images showed a hip joint effusion with a continuous bursa extending medially and displacing the obturator externus inferiorly. At surgery, a bursa was seen displacing the obturator externus muscle inferiorly and originating from the posteroinferior aspect of the hip joint in all eight patients. Results of pathologic analysis confirmed disease identical to the primary hip abnormality in all eight patients. CONCLUSION: The obturator externus bursa is a potential posteroinferior communication of the hip joint capsule, can be a site of disease spread from the hip joint, and can be accurately identified with MR imaging. PMID- 12775850 TI - Wavelet compression of low-dose chest CT data: effect on lung nodule detection. AB - PURPOSE: To assess the effect of using a lossy Joint Photographic Experts Group standard for wavelet image compression, JPEG2000, on pulmonary nodule detection at low-dose computed tomography (CT). MATERIALS AND METHODS: One hundred sets of lung CT data ("cases") were compressed to 30:1, 20:1, and 10:1 levels by using a wavelet-based JPEG2000 method, resulting in 400 test cases. Each case consisted of nine 1.25-mm sections that had been obtained with 20-40 mAs. Four thoracic radiologists independently interpreted the test case images. Performance was measured by using area under the receiver operating characteristic (ROC) curve (Az) and conventional sensitivity and specificity analyses. RESULTS: There were 51 cases with and 49 without lung nodules. Az values were 0.984, 0.988, 0.972, 0.921, respectively, for original and 10:1, 20:1, and 30:1 compressed images. Az values decreased significantly at 30:1 (P =.014) but not at 10:1 compression, with a trend toward significant decrease at 20:1 (P =.051). Specificity values were unaffected by compression (>98.0% at all compression levels). Sensitivity values were 86.3% (176 of 204 test cases with nodules), 77.9% (159 of 204 cases), 76.5% (156 of 204 cases), and 70.1% (143 of 204 cases), respectively, for original and 10:1, 20:1, and 30:1 compressed images. Results of logistic regression model analysis confirmed the significant effects of compression rate and nodule attenuation, size, and location on sensitivity (P <.05). CONCLUSION: While no reduction in nodule detection at 10:1 compression levels was demonstrated by using ROC analysis, a significant decrease in sensitivity was identified. Further investigation is needed before widespread use of image compression technology in low-dose chest CT can be recommended. PMID- 12775851 TI - Invasive ductal breast carcinoma response to neoadjuvant chemotherapy: noninvasive monitoring with functional MR imaging pilot study. AB - PURPOSE: To investigate if the extraction flow product (EFP), as determined on dynamic contrast material-enhanced magnetic resonance (MR) images, could be a potential marker of tumor response to neoadjuvant chemotherapy in patients with locally advanced breast cancer. MATERIALS AND METHODS: Fourteen women with proven breast cancer underwent MR imaging prior to and following neoadjuvant chemotherapy. Dynamic gradient-echo and echo-planar MR images were acquired before and after injection of gadopentetate dimeglumine. Precontrast T1s were measured before EFP maps were calculated by using a multicompartmental model. Mean EFP (EFPmean) and distribution analysis of EFP (EFPcount) were measured in tumors before and after neoadjuvant chemotherapy and were compared with tumor response at MR imaging. The significance of the difference in EFP values between the responders and nonresponders was calculated with a two-tailed Student t test. RESULTS: EFPmean after neoadjuvant chemotherapy in partial responders and nonresponders was 33 mL x 100 g-1 x min-1 +/- 9.8 and 54.2 mL x 100 g-1 x min-1 +/- 10.3, respectively (P <.005). EFPmean decreased after neoadjuvant chemotherapy in the responders and nonresponders by 37% +/- 30 and -5% +/- 35, respectively (P >.05). An increase in EFPmean values was observed only in nonresponders who received taxanes. For regimens without taxanes, EFPmean decreased regardless of the morphologic response. EFPcount decreased for all the responders by 77% +/- 33 and increased for all the nonresponders by 45% +/- 68 (P <.02). CONCLUSION: EFPcount appears to provide functional information regarding changes in tumor angiogenesis due to neoadjuvant chemotherapy. Functional MR imaging of the breast may be useful in monitoring tumor response to neoadjuvant chemotherapy. PMID- 12775852 TI - Pulmonary embolism guidelines: will they work? PMID- 12775853 TI - Nicotine replacement therapy in smoking cessation. PMID- 12775854 TI - ADAM 33: just another asthma gene or a breakthrough in understanding the origins of bronchial hyperresponsiveness? PMID- 12775857 TI - Clinical trial comparing nicotine replacement therapy (NRT) plus brief counselling, brief counselling alone, and minimal intervention on smoking cessation in hospital inpatients. AB - BACKGROUND: Guidelines recommend that smoking cessation interventions are offered in all clinical settings to all smokers willing to make a quit attempt. Since the effectiveness of routine provision of behavioural counselling and nicotine replacement therapy (NRT) to smokers admitted to hospital has not been established, a randomised controlled trial of these interventions given together compared with counselling alone or minimal intervention was performed in hospital inpatients. METHODS: Medical and surgical inpatients who were current smokers at the time of admission were randomised to receive either usual care (no additional advice at admission), counselling alone (20 minute intervention with written materials), or NRT plus counselling (counselling intervention with a 6 week course of NRT). Continuous and point prevalence abstinence from smoking (validated by exhaled carbon monoxide <10 ppm) was measured at discharge from hospital and at 3 and 12 months, and self-reported reduction in cigarette consumption in smokers was assessed at 3 and 12 months. RESULTS: 274 inpatient smokers were enrolled. Abstinence was higher in the NRT plus counselling group (n=91) than in the counselling alone (n=91) or usual care (n=92) groups. The difference between the groups was significant for validated point prevalence abstinence at discharge (55%, 43%, 37% respectively, p=0.045) and at 12 months (17%, 6%, 8%, p=0.03). The respective differences in continuous validated abstinence at 12 months were 11%, 4%, 8% (p=0.25). There was no significant difference between counselling alone and usual care, or in reduction in cigarette consumption between the treatment groups. CONCLUSIONS: NRT given with brief counselling to hospital inpatients is an effective routine smoking cessation intervention. PMID- 12775858 TI - Primary prevention of asthma and atopy during childhood by allergen avoidance in infancy: a randomised controlled study. AB - BACKGROUND: Recent increases in the prevalence of asthma and atopy emphasise the need for devising effective methods for primary prevention in children at high risk of atopy. METHOD: A birth cohort of genetically at risk infants was recruited in 1990 to a randomised controlled study. Allergen avoidance measures were instituted from birth in the prophylactic group (n=58). Infants were either breast fed with mother on a low allergen diet or given an extensively hydrolysed formula. Exposure to house dust mite was reduced by the use of an acaricide and mattress covers. The control group (n=62) followed standard advice as normally given by the health visitors. At age 8, all 120 children completed a questionnaire and 110 (92%) had all assessments (skin prick test, spirometry, and bronchial challenges). RESULTS: In the prophylactic group eight children (13.8%) had current wheeze compared with 17 (27.4%) in the control group (p=0.08). Respective figures were eight (13.8%) and 20 (32.3%) for nocturnal cough (p=0.02) and 11 of 55 (20.0%) and 29 of 62 (46.8%) for atopy (p=0.003). After adjusting for confounding variables, the prophylactic group was found to be at a significantly reduced risk for current wheeze (odds ratio (OR) 0.26 (95% confidence interval (CI) 0.07 to 0.96)), nocturnal cough (OR 0.22 (95% CI 0.06 to 0.83)), asthma as defined by wheeze and bronchial hyperresponsiveness (OR 0.11 (95% CI 0.01 to 1.02)), and atopy (OR 0.21 (95% CI 0.07 to 0.62)). CONCLUSION: Strict allergen avoidance in infancy in high risk children reduces the development of allergic sensitisation to house dust mite. Our results suggest that this may prevent some cases of childhood asthma. PMID- 12775859 TI - Exhaled nitric oxide rather than lung function distinguishes preschool children with probable asthma. AB - BACKGROUND: Respiratory function and airway inflammation can be evaluated in preschool children with special techniques, but their relative power in identifying young children with asthma has not been studied. This study was undertaken to compare the value of exhaled nitric oxide (FE(NO)), baseline lung function, and bronchodilator responsiveness in identifying children with newly detected probable asthma. METHODS: Ninety six preschool children (age 3.8-7.5 years) with asthmatic symptoms or history and 62 age matched healthy non-atopic controls were studied. FE(NO) was measured with the standard online single exhalation technique, and baseline lung function and bronchodilator responsiveness were measured using impulse oscillometry (IOS). RESULTS: Children with probable asthma (n=21), characterised by recent recurrent wheeze, had a significantly higher mean (SE) concentration of FE(NO) than controls (22.1 (3.4) ppb v 5.3 (0.4) ppb; mean difference 16.8 ppb, 95% CI 12.0 to 21.5) and also had higher baseline respiratory resistance, lower reactance, and larger bronchodilator responses expressed as the change in resistance after inhalation of salbutamol. Children with chronic cough only (n=46) also had significantly raised mean FE(NO) (9.2 (1.5) ppb; mean difference 3.9 ppb, 95% CI 0.8 to 7.0) but their lung function was not significantly reduced. Children on inhaled steroids due to previously diagnosed asthma (n=29) differed from the controls only in their baseline lung function. The analysis of receiver operating characteristics (ROC) showed that FE(NO) provided the best power for discriminating between children with probable asthma and healthy controls, with a sensitivity of 86% and specificity of 92% at the cut off level of 1.5 SD above predicted. CONCLUSIONS: FE(NO) is superior to baseline respiratory function and bronchodilator responsiveness in identifying preschool children with probable asthma. The results emphasise the presence of airway inflammation in the early stages of asthma, even in young children. PMID- 12775860 TI - Sustained reduction in bronchial hyperresponsiveness with inhaled fluticasone propionate within three days in mild asthma: time course after onset and cessation of treatment. AB - BACKGROUND: Bronchial hyperresponsiveness (BHR) is characteristic of asthmatic airways, is induced by airway inflammation, and is reduced by inhaled corticosteroids (ICS). The time course for the onset and cessation of the effect of ICS on BHR is unclear. The effect of inhaled fluticasone propionate (FP) on BHR in patients with mild persistent asthma was assessed using time intervals of hours, days and weeks. METHODS: Twenty six asthmatic patients aged 21-59 years were selected for this randomised, double blind, parallel group study. The effect of 250 micro g inhaled FP (MDI) administered twice daily was compared with that of placebo on BHR assessed using a dosimetric histamine challenge method. The dose of histamine inducing a decrease in forced expiratory volume in 1 second (FEV(1)) by 15% (PD(15)FEV(1)) was measured before and 6, 12, 24 and 72 hours, and 2, 4 and 6 weeks after starting treatment, and 48 hours, 1 week and 2 weeks after cessation of treatment. Doubling doses of changes in PD(15)FEV(1) were calculated and area under the curve (AUC) statistics were used to summarise the information from individual response curves. RESULTS: The increase in PD(15)FEV(1) from baseline was greater in the FP group than in the placebo group; the difference achieved significance within 72 hours and remained significant until the end of treatment. In the FP group PD(15)FEV(1) was 1.85-2.07 doubling doses above baseline between 72 hours and 6 weeks after starting treatment. BHR increased significantly within 2 weeks after cessation of FP treatment. CONCLUSIONS: A sustained reduction in BHR to histamine in patients with mild asthma was achieved within 3 days of starting treatment with FP at a daily dose of 500 micro g. The effect tapered within 2 weeks of cessation of treatment. PMID- 12775861 TI - Cysteinyl leukotrienes and 8-isoprostane in exhaled breath condensate of children with asthma exacerbations. AB - BACKGROUND: Cysteinyl leukotrienes (Cys-LTs) and isoprostanes are inflammatory metabolites derived from arachidonic acid whose levels are increased in the airways of asthmatic patients. Isoprostanes are relatively stable and specific for lipid peroxidation, which makes them potentially reliable biomarkers for oxidative stress. A study was undertaken to evaluate the effect of a course of oral steroids on Cys-LT and 8-isoprostane levels in exhaled breath condensate of children with an asthma exacerbation. METHODS: Exhaled breath condensate was collected and fractional exhaled nitric oxide (FE(NO)) and spirometric parameters were measured before and after a 5 day course of oral prednisone (1 mg/kg/day) in 15 asthmatic children with an asthma exacerbation. Cys-LT and 8-isoprostane concentrations were measured using an enzyme immunoassay. FE(NO) was measured using a chemiluminescence analyser. Exhaled breath condensate was also collected from 10 healthy children. RESULTS: Before prednisone treatment both Cys-LT and 8 isoprostane concentrations were higher in asthmatic subjects (Cys-LTs, 12.7 pg/ml (IQR 5.4-15.6); 8-isoprostane, 12.0 pg/ml (9.4-29.5)) than in healthy children (Cys-LTs, 4.3 pg/ml (2.0-5.7), p=0.002; 8-isoprostane, 2.6 pg/ml (2.1-3.0), p<0.001). After prednisone treatment there was a significant decrease in both Cys LT (5.2 pg/ml (3.9-8.8), p=0.005) and 8-isoprostane (8.4 pg/ml (5.4-11.6), p=0.04) concentrations, but 8-isoprostane levels remained higher than in controls (p<0.001). FE(NO) levels, which fell significantly after prednisone treatment (p<0.001), did not correlate significantly with either Cys-LT or 8-isoprostane concentrations. CONCLUSION: After a 5 day course of oral prednisone there is a reduction in Cys-LT and 8-isoprostane levels in EBC of children with an asthma exacerbation, although 8-isoprostane levels remain higher than in controls. This finding suggests that corticosteroids may not be fully effective in reducing oxidative stress in children with an exacerbation of asthma. PMID- 12775856 TI - British Thoracic Society guidelines for the management of suspected acute pulmonary embolism. PMID- 12775863 TI - Selection of patients for lung volume reduction surgery using a power law analysis of the computed tomographic scan. AB - BACKGROUND: A study was undertaken to test the hypothesis that patients respond better to lung volume reduction surgery (LVRS) if their emphysema is confluent and predominantly located in the upper lobes. METHODS: A density mask analysis was used to identify voxels inflated beyond 10.2 ml gas/g tissue (-910 HU) on preoperative and postoperative CT scans from patients receiving LVRS. These hyperinflated regions were considered to represent emphysematous lesions. A power law analysis was used to determine the relationship between the number (K) and size (A) of the emphysematous lesions in the whole lung and two anatomical regions using the power law equation Y=KA(-D). RESULTS: The analysis showed a positive correlation between the change in the power law exponent (D) and the change in exercise (Watts) after surgery (r=0.47, p=0.03). There was also a negative correlation between the power law exponent D in the upper region of the lung preoperatively and the change in exercise following surgery (r=-0.60, p<0.05). CONCLUSIONS: These results confirm that patients with large upper lobe lesions respond better to LVRS than patients with small uniformly distributed disease. Power law analysis of lung CT scans provides a quantitative method for determining the extent and location of emphysema within the lungs of patients with COPD. PMID- 12775865 TI - Health status measurement: sensitivity of the self-reported Chronic Respiratory Questionnaire (CRQ-SR) in pulmonary rehabilitation. AB - BACKGROUND: A self-reported CRQ (CRQ-SR) has recently been developed and found to be a reproducible and reliable measure of health status. This study explores both the sensitivity of the CRQ-SR and relative sensitivity compared with the conventional interviewer led CRQ (CRQ-IL) in patients undergoing pulmonary rehabilitation. METHODS: Eighty patients with stable chronic obstructive pulmonary disease who had been referred for pulmonary rehabilitation completed the CRQ-SR at initial assessment and at the end of the 7 week programme. A further 35 patients completed both the CRQ-SR and the CRQ-IL, administered 1 week apart, before starting rehabilitation and again at the end of the programme. RESULTS: There were large statistically and clinically significant changes in mean score per dimension following rehabilitation in all dimensions of the CRQ-SR (dyspnoea mean difference 0.87 (95% CI 0.61 to 1.14); fatigue 0.76 (0.53 to 1.0); emotion 0.60 (0.35 to 0.86); mastery 0.76 (0.52 to 1.0); p<0.001). Similar results were found in the comparison of the sensitivity of the CRQ-SR and the CRQ IL, with large changes in mean score per dimension following rehabilitation for both versions of the questionnaire (p<0.005). No significant differences were seen in the magnitude of change between the two formats of the questionnaire (p>0.05). CONCLUSION: The self-reported CRQ is as sensitive to change as the interviewer led CRQ in patients undergoing pulmonary rehabilitation but has the advantage of being less time consuming to administer. PMID- 12775866 TI - Increased interleukin-13 expression in patients with sarcoidosis. AB - BACKGROUND: Sarcoidosis is a systemic granulomatous disorder of unknown origin. Lymphocytic inflammation is dominated by expression of Th1 type cytokines such as tumour necrosis factor alpha (TNFalpha). Interleukin 13 (IL-13) is a Th2 cytokine which is expressed by CD4+ T cells and has been shown to suppress TNFalpha in human blood monocytes. The role of IL-13 as a possible anti-inflammatory cytokine in sarcoidosis was investigated. METHODS: mRNA expression of IL-13, IL-4, IL-10, and TNFalpha in bronchoalveolar lavage (BAL) fluid cells and peripheral mononuclear blood cells (PBM) of 18 patients with sarcoidosis and nine healthy controls was assessed using RT-PCR. In addition, IL-13 protein levels in BAL cell culture supernatants from 12 patients and all controls were measured and immunocytochemistry of IL-13 protein was performed in BAL fluid cells of eight patients. TNFalpha concentrations were measured with and without stimulation with recombinant human (rh) IL-13, rhIL-10, and lipopolysaccharide (LPS). RESULTS: IL 13 mRNA expression was significantly increased in BAL cells and PBM of patients compared with controls (p<0.05). No significant difference was found in IL-4 mRNA or IL-10 mRNA expression in BAL fluid cells or PBM between the two groups. TNFalpha mRNA expression was significantly higher in BAL fluid cells of patients than controls (p<0.05). IL-13 protein levels in BAL cell culture supernatants were slightly raised in half the patients investigated but in only two controls. Immunocytochemistry detected IL-13 protein in alveolar macrophages of patients. IL-13 led to decreased TNFalpha concentrations (p<0.05). CONCLUSIONS: IL-13 expression is increased in BAL cells and PBM in sarcoidosis and IL-13 is secreted from BAL cells. Alveolar macrophages may be the cellular source. These data suggest that IL-13 might have an anti-inflammatory effect by acting on TNFalpha. PMID- 12775867 TI - Identification of airborne dissemination of epidemic multiresistant strains of Pseudomonas aeruginosa at a CF centre during a cross infection outbreak. AB - BACKGROUND: Chronic Pseudomonas aeruginosa infection is a major cause of morbidity and mortality for individuals with cystic fibrosis (CF). P aeruginosa cross infection outbreaks have recently been reported at CF holiday camps and specialist centres. The mechanism of cross infection is unknown. A study was performed to look for the presence of epidemic strains of P aeruginosa in the environment of a CF centre during a cross infection outbreak and to examine their potential modes of spread between patients. METHODS: Microbiological sampling of the environment of the CF facility was performed, including room air sampling. Individual P aeruginosa strains were identified by bacterial fingerprinting. The typing patterns were compared with those of epidemic strains responsible for cross infection among the patients. RESULTS: Epidemic P aeruginosa strains were isolated from room air when patients performed spirometric tests, nebulisation, and airway clearance, but were not present in other areas of the inanimate environment of the CF centre. CONCLUSIONS: Aerosol dissemination may be the most important factor in patient-to-patient spread of epidemic strains of P aeruginosa during recent cross infection outbreaks at adult CF centres. PMID- 12775868 TI - Comparison of airway immunopathology of eosinophilic bronchitis and asthma. AB - BACKGROUND: Eosinophilic bronchitis is a condition characterised by a corticosteroid responsive cough, sputum eosinophilia, and normal tests of variable airflow obstruction and airway responsiveness. We performed a detailed comparative immunopathological study to test the hypothesis that the different airway function in patients with eosinophilic bronchitis and asthma reflects differences in the nature of the lower airway inflammatory response. METHODS: Exhaled nitric oxide was measured and induced sputum, bronchoscopy, bronchial wash (BW), bronchoalveolar lavage (BAL), and bronchial biopsy were performed in 16 subjects with eosinophilic bronchitis, 15 with asthma, and 14 normal controls. RESULTS: Both eosinophilic bronchitis and asthma were characterised by an induced sputum, BW and BAL eosinophilia, an increased number of epithelial and subepithelial eosinophils, and increased reticular basement membrane thickness. The median concentration of exhaled nitric oxide was higher in those with eosinophilic bronchitis (12 ppb) or asthma (8.5 ppb) than normal controls (2 ppb) (95% CI of the difference 5 to 16, p<0.0001 and 2 to 11.3, p=0.004, respectively). There were no group differences in epithelial integrity or the number of subepithelial T lymphocytes, mast cells or macrophages. CONCLUSION: With the exception of our previously reported association of smooth muscle mast cell infiltration with asthma, the immunopathology of eosinophilic bronchitis and asthma are similar which suggests that eosinophilic airway inflammation, increased exhaled nitric oxide, and increased basement membrane thickening are regulated independently of airway hyperresponsiveness. PMID- 12775870 TI - Respiratory symptoms in patients with treated hypothyroidism and inflammatory bowel disease. AB - BACKGROUND: Patients with idiopathic chronic cough and unexplained airflow obstruction in non-smokers have been shown to have an increased prevalence of hypothyroidism and other organ specific autoimmune disorders. Whether patients with hypothyroidism have an increased prevalence of respiratory symptoms is unknown. METHODS: The prevalence of respiratory symptoms was assessed in 124 patients with treated hypothyroidism recruited from primary and secondary care, 64 outpatients with inflammatory bowel disease, and 1346 control adults recruited randomly from the electoral register in a case-control study. Respiratory symptoms and smoking history were assessed by a respiratory symptom questionnaire. RESULTS: After adjustment for age, sex and smoking, symptoms of breathlessness and sputum production were more prevalent in both patient populations than in controls (odds ratios for hypothyroidism and inflammatory bowel disease; breathlessness: 3.1 (95% CI 2.1 to 4.6) and 3.4 (95% CI 2.0 to 6.0), respectively; sputum production: 2.7 (95% CI 1.6 to 4.5) and 2.5 (95% CI 1.2 to 5.0), respectively). Cough during the day and night was significantly more prevalent in patients with hypothyroidism (1.8 (95% CI 1.2 to 2.9)) and approached significance in those with inflammatory bowel disease (1.8 (95% CI 1.0 to 3.4)). Wheeze and nocturnal cough were no more prevalent in either disease population than in controls. CONCLUSION: There is a significantly increased prevalence of respiratory symptoms in patients with hypothyroidism or inflammatory bowel disease compared with controls recruited from the general population. Further work is required to determine whether similar differences are seen in comparison with hospital based controls. These findings support the hypothesis that there is a link between autoimmune hypothyroidism and respiratory disease. PMID- 12775872 TI - Chronic obstructive pulmonary disease * 9: management of ventilatory failure in COPD. AB - The management of respiratory failure during acute exacerbations of COPD and during chronic stable COPD is reviewed and the role of non-invasive and invasive mechanical ventilation is discussed. PMID- 12775875 TI - The pulmonary physician in critical care * illustrative case 4: neuromusculoskeletal disorders. AB - The case history is presented of a patient admitted to the ICU with ventilatory insufficiency following thoracotomy for thymic resection. The role of non invasive ventilation for weaning in patients following phrenic nerve injury is discussed. PMID- 12775876 TI - Pulmonary vein thrombosis after bilobectomy and development of collateral circulation. AB - Pulmonary vein thrombosis is a rare but potentially life threatening complication following lobectomy or bilobectomy. We present a case of right upper pulmonary vein thrombosis after a middle and lower lobectomy diagnosed at transoesophageal echocardiography. The patient was treated with antibiotics and anticoagulation with good recovery. Pulmonary angiography was performed 35 days after surgery and revealed the venous return of the right lung through the intercostal veins. Despite double venous drainage of the lungs consisting of bronchial and pulmonary veins, pulmonary to systemic collaterals following pulmonary vein thrombosis have not previously been reported. The development of this shunt can prevent gangrene, and surgical resection of the lung segment involved can be avoided. PMID- 12775874 TI - The pulmonary physician in critical care * illustrative case 3: pulmonary vasculitis. AB - The case history of a patient admitted to the ICU with severe hypoxic respiratory failure later diagnosed as Wegener's granulomatosis is presented. The diagnosis and management of patients with suspected pulmonary vasculitis is discussed. PMID- 12775877 TI - Sarcoidosis presenting as upper extremity venous thrombosis. PMID- 12775878 TI - Genotype-phenotype correlations in PCD patients carrying DNAH5 mutations. PMID- 12775879 TI - Sleep disordered breathing in an adult with cherubism. PMID- 12775880 TI - Long-term angiographic recurrences after selective endovascular treatment of aneurysms with detachable coils. AB - BACKGROUND AND PURPOSE: Our aim in this study was to assess the incidence and determining factors of angiographic recurrences after endovascular treatment of aneurysms. METHODS: A retrospective analysis of all patients with selective endosaccular coil occlusion of intracranial aneurysms prospectively collected from 1992 to 2002 was performed. There were 501 aneurysms in 466 patients (mean+/ SD age, 54.20+/-12.54 years; 74% female). Aneurysms were acutely ruptured (54.1%) or unruptured (45.9%). Mean+/-SD aneurysm size was 9.67+/-5.91 mm with a 4.31+/ 1.97-mm neck. The most frequent sites were basilar bifurcation (27.7%) and carotid ophthalmic (18.0%) aneurysms. Recurrences were subjectively divided into minor and major (ideally necessitating re-treatment). The most significant predictors of angiographic recurrence were determined by logistic regression. These results were confirmed by chi2, t tests, or ANOVAs followed, when appropriate, by Tukey's contrasts. RESULTS: Short-term (< or =1 year) follow-up angiograms were available in 353 aneurysms (70.5%) and long-term (>1 year) follow up angiograms, in 277 (55%), for a total of 383 (76.5%) followed up. Recurrences were found in 33.6% of treated aneurysms that were followed up and that appeared at a mean+/-SD time of 12.31+/-11.33 months after treatment. Major recurrences presented in 20.7% and appeared at a mean of 16.49+/-15.93 months. Three patients (0.8%) bled during a mean clinical follow-up period of 31.32+/-24.96 months. Variables determined to be significant predictors (P<0.05) of a recurrence included aneurysm size > or =10 mm, treatment during the acute phase of rupture, incomplete initial occlusions, and duration of follow-up. CONCLUSIONS: Long-term monitoring of patients treated by endosaccular coiling is mandatory. PMID- 12775882 TI - Abdominal obesity and risk of ischemic stroke: the Northern Manhattan Stroke Study. AB - BACKGROUND AND PURPOSE: Obesity is well recognized as a risk factor for coronary heart disease and mortality. The relationship between abdominal obesity and ischemic stroke remains less clear. Our aim was to evaluate abdominal obesity as an independent risk factor for ischemic stroke in a multiethnic community. METHODS: A population-based, incident case-control study was conducted July 1993 through June 1997 in northern Manhattan, New York, NY. Cases (n=576) of first ischemic stroke (66% >or=BORDER="0">65 years of age; 56% women; 17% whites; 26% blacks; 55% Hispanics) were enrolled and matched by age, sex, and race-ethnicity to stroke-free community controls (n=1142). All subjects were interviewed and examined and had measurements of waist-to-hip ratio (WHR). Odds ratios (ORs) of ischemic stroke were calculated with gender-specific quartiles (GQs) and gender specific medians of WHR adjusted for stroke risk factors and body mass index (BMI). RESULTS: Compared with the first quartile, the third and fourth quartiles of WHR had an increased risk of stroke (GQ3: OR, 2.4; 95% CI, 1.5 to 3.9; GQ4: OR, 3.0; 95% CI, 1.8 to 4.8) adjusted for other risk factors and BMI. Those with WHR equal to or greater than the median had an overall OR of 3.0 (95% CI, 2.1 to 4.2) for ischemic stroke even after adjustment for other risk factors and BMI. Increased WHR was associated with a greater risk of stroke in men and women and in all race-ethnic groups. The effect of WHR was stronger among younger persons (test for heterogeneity, P<0.0002) (<65 years of age: OR, 4.4; 95%CI, 2.2 to 9.0; >or=65 years of age: OR, 2.2; 95% CI, 1.4 to 3.2). WHR was associated with an increased risk among those with and without large-artery atherosclerotic stroke. CONCLUSIONS: Abdominal obesity is an independent, potent risk factor for ischemic stroke in all race-ethnic groups. It is a stronger risk factor than BMI and has a greater effect among younger persons. Prevention of obesity and weight reduction need greater emphasis in stroke prevention programs. PMID- 12775881 TI - Molecular determinants of the prothrombogenic and inflammatory phenotype assumed by the postischemic cerebral microcirculation. AB - BACKGROUND AND PURPOSE: Circulating blood cells have been implicated in the pathogenesis of cerebral ischemia/reperfusion (I/R) injury and stroke. The objective of this study was to define the magnitude and molecular determinants of the platelet- and leukocyte-endothelial cell adhesive interactions induced by I/R in the mouse brain. METHODS: Bilateral common carotid artery occlusion was induced for 1 hour in C57BL/6 mice, followed by either 40 minutes or 4 hours of reperfusion. Fluorescent platelets were administered intravenously, and the frontal brain surface was observed with intravital fluorescence microscopy. Leukocyte-endothelial cell adhesion was monitored with the use of rhodamine-6G. RESULTS: Ischemia followed by 40 minutes of reperfusion resulted in the rolling (125.1+/-23.6/mm2) and firm adhesion (109.5+/-25.8/mm2) of leukocytes but not platelets in venules. However, with 4 hours of reperfusion, rolling (138.8+/ 24.6/mm2) and firm adhesion (153.7+/-22.3/mm2) of platelets were detected, and this was accompanied by a more intense recruitment of rolling (374.5+/-54.6/mm2) and adherent (445.2+/-57.1/mm2) leukocytes. In mice deficient in either P selectin (P-selectin-/-) or intercellular adhesion molecule-1 (ICAM-1) (ICAM-1-/ ), the I/R-induced platelet-endothelial cell (by 80% and 60%, respectively) and leukocyte-endothelial cell (by 84% and 78%, respectively) interactions were significantly blunted compared with those of wild-type mice. CONCLUSIONS: These findings indicate that I/R promotes the adhesion of both platelets and leukocytes in cerebral venules, with the accumulation of adherent leukocytes preceding the recruitment of platelets. Both P-selectin and ICAM-1 contribute to the inflammatory and prothrombogenic state induced by cerebral I/R. PMID- 12775883 TI - Comparison of psychometric properties of three mobility measures for patients with stroke. AB - BACKGROUND AND PURPOSE: This study compared the validity, responsiveness, and interrater reliability of 3 mobility measures in stroke patients from the acute stage up to 180 days after stroke onset. The 3 measures were the Rivermead Mobility Index (RMI), a modified RMI (MRMI), and the Mobility Subscale of the Stroke Rehabilitation Assessment of Movement (STREAM). METHODS: The validity and responsiveness of the 3 mobility measures were prospectively examined by monitoring 57 stroke patients with the measures and the Barthel Index at 14, 30, 90, and 180 days after stroke onset. Two individual raters used the 3 measures to evaluate a different sample of 40 patients on 2 separate occasions to determine the interrater reliability. RESULTS: The Spearman rho between STREAM and MRMI was >or=BORDER="0">0.92; the intraclass correlation coefficient (ICC, a measure of agreement) between them was >or=0.89, indicating high concurrent validity of both measures. RMI showed a moderate to high relationship and agreement with STREAM and MRMI (rho>or=0.78, ICC>or=0.5). Responsiveness of the 3 measures was high before 90 days after stroke onset (standardized response mean >or=0.83) and low at 90 to 180 days after stroke onset (0.2or=BORDER="0">0.92). CONCLUSIONS: All 3 measures examined showed acceptable levels of reliability, validity, and responsiveness in stroke patients. The psychometric characteristics of STREAM were slightly superior to those of the other 2 measures among our patients. We prefer and recommend STREAM for measuring mobility disability in stroke patients. PMID- 12775884 TI - Central retinal artery Doppler flow parameters reflect the severity of cerebral small-vessel disease. AB - BACKGROUND AND PURPOSE: We investigated the usefulness of central retinal artery (CRA) Doppler flowmetry in patients with cerebral small-vessel disease (SVD). METHODS: CRA Doppler flowmetry was performed in 103 SVD patients who underwent MRI. Sixty-four adjusted control subjects were also registered. We assessed average CRA flow parameter values for both eyes with the clinical and MRI findings. RESULTS: Each Doppler flowmetry was performed within 5 minutes. Patients with SVD had significantly lower end-diastolic and mean velocities of the CRA than control subjects; they also had higher pulsatility and resistive indexes. Multivariate analysis showed that the number of small infarcts was an independent predictor of peak systolic and mean velocities. Grade of periventricular hyperintensities was an additional independent predictor of peak systolic and mean velocities, whereas the number of small infarcts was predictive of end-diastolic velocity. CONCLUSIONS: Flow parameters may be useful for the quantitative assessment of SVD severity. PMID- 12775885 TI - Association of methylenetetrahydrofolate reductase gene polymorphism with carotid atherosclerosis depending on smoking status in a Japanese general population. AB - BACKGROUND AND PURPOSE: The association of the methylenetetrahydrofolate reductase gene (MTHFR) with carotid atherosclerosis remains inconsistent. This may be due to small sample size and inappropriate analysis. We investigated the association of C677T/MTHFR with blood pressure and carotid atherosclerosis in a Japanese general population. METHODS: Subjects (30 to 89 years of age; 1693 women, 1554 men) who gave informed consent were randomly selected from a general population in Suita, Japan. MTHFR genotypes were determined by TaqMan polymerase chain reaction. Carotid atherosclerosis was evaluated by high-resolution ultrasonography with atherosclerotic indexes of intimal-medial thickness (IMT), maximum IMT in the common carotid artery (CCA), plaque score, and stenosis (>50%). RESULTS: Age-adjusted diastolic blood pressure was significantly higher in women with the TT genotype than in those with the CC genotype. In a recessive model (CC+CT versus TT), all adjusted odds ratios for hypertension and >50% stenosis in women were 1.42 and 3.42 (95% confidence intervals, 1.01 to 1.99 and 1.23 to 9.53), respectively. In women, maximum IMT in CCA for smokers with the TT genotype was significantly higher than for smokers with the CC genotype and nonsmokers with the TT genotype (P<0.05). CONCLUSIONS: Our study suggests that the MTHFR TT genotype is a risk factor for hypertension and carotid stenosis in women. Significant interactions between C677T/MTHFR and smoking on maximum IMT in CCA were observed in women but not in men. Smoking cessation for subjects with the TT genotype is important in the prevention of cerebrovascular disease. PMID- 12775886 TI - Endoglin is not a major susceptibility gene for intracranial aneurysm among Japanese. AB - BACKGROUND AND PURPOSE: A 6-base insertion (6bINS) polymorphism in intron 7 of the endoglin gene (ENG), which codes for a component of the transforming growth factor-beta receptor complex, was reported to be associated with intracranial aneurysm (IA) in a Japanese population. A recent report using a white population could not replicate the association. We tested for this association with high statistical power in our independent Japanese subjects and evaluated the linkage between markers on chromosome 9, which contains ENG, and IA. METHODS: The sample for the linkage study comprised 179 individuals with IA in 85 nuclear families, with 104 possible affected sibpairs. For the association study of the 6bINS polymorphism and 4 single nucleotide polymorphisms (SNPs) in ENG, 172 Japanese patients with IA and 192 control subjects were examined. RESULTS: There was no evidence of linkage in the vicinity of ENG by analysis of affected sibpairs. The allele frequency of the 6bINS polymorphism was 104 of 344 (30.2%) in the total IA group and 122 of 382 (31.9%) in the control group. The statistical difference in allele frequency between the 2 groups was not significant (chi2=0.245, df=1, P=0.620). The power of the present association study was 98.3% at a significance level of 0.05 on the basis of the allele frequencies in the previous study. In addition, no associations between the 4 SNPs in ENG and IA were detected. CONCLUSIONS: We provide evidence that there is no association between the 6bINS polymorphism or 4 SNPs in ENG and IA and that there is no linkage between the ENG locus and IA, indicating that ENG is not a major susceptibility gene for IA in Japanese. PMID- 12775887 TI - Smokeless tobacco as a possible risk factor for stroke in men: a nested case control study. AB - BACKGROUND AND PURPOSE: Snuff and other forms of smokeless tobacco are widely used in some populations. Whereas the detrimental cardiovascular effects of smoking are well documented, possible health hazards associated with the use of smokeless tobacco remain controversial. The purpose of this study was to explore whether the use of snuff, a smokeless tobacco product, increases the risk of stroke in men. METHODS: In a nested case-control study (1 case and 2 matched controls without cardiovascular disease), information on tobacco habits was collected through population risk factor surveys. During follow-up, first-ever fatal and nonfatal strokes occurring among 25- to 74-year-old participants were identified in a population-based stroke register. The study was restricted to men (276 cases, 551 controls). RESULTS: The unadjusted odds ratio for stroke in regular cigarette smokers compared with men who never used tobacco was 2.21 (95% CI, 1.29 to 3.79). When never-smoking regular snuff dippers (excluding ex smokers) were compared with men who never used tobacco, the unadjusted odds ratio was 1.05 (95% CI, 0.37 to 2.94). The odds ratio for never-snuffing smokers versus never-smoking snuff users was 2.90 (95% CI, 0.92 to 9.1). After adjustment for multiple cardiovascular risk factors, the odds ratio for having a stroke was 1.74 (95% CI, 0.85 to 3.54) in regular smokers and 0.87 (95% CI, 0.41 to 1.83) in regular snuff users. CONCLUSIONS: Whereas regular smoking doubles the risk of stroke in men, snuff use is not associated with any apparent excess risk. Chemical moieties produced by burning tobacco are probably the most important contributors to smokers' excess risk of atherothrombotic disease, including stroke. PMID- 12775888 TI - Effect of carotid endarterectomy on primary collateral blood flow in patients with severe carotid artery lesions. AB - BACKGROUND AND PURPOSE: In patients with severe obstruction of the internal carotid artery (ICA), it is recognized that the preoperative failure to visualize collaterals of the circle of Willis increases the risk of hemispheric ischemia before, during, and after carotid endarterectomy (CEA). The purpose of the present study was to assess the effect of CEA on the anatomy and function of the circle of Willis. METHODS: Time-of-flight and phase-contrast MR angiography were used to study changes in vessel diameter and collateral flow of the circle of Willis in 48 patients with 70% to 99% ICA stenosis before and after CEA. RESULTS: In patients with unilateral ICA stenosis, all preoperative vessel diameters on both sides of the circle of Willis were larger than in control subjects. All demonstrated a significant diameter decrease to reach normal values after CEA. Furthermore, preoperative collateral flow patterns normalized after CEA (P=0.03). In patients with stenosis and contralateral ICA occlusion, CEA resulted in a significant increase in the prevalence of collateral flow via the anterior communicating artery (33% to 83%, P<0.01) and a significant increase in diameter of both A1 segments (P<0.05) in patients in whom collateral flow developed after CEA. CONCLUSIONS: CEA reduces the caliber of compensatory collateral channels to normal levels by MR angiography measurements in the presence of severe unilateral stenosis; when the opposite side is occluded and the stenosis is removed ipsilaterally, a greater amount of compensatory collateral circulation can be measured on both the occluded side and the fully opened side. PMID- 12775889 TI - Neither arm nor face warming reduces the shivering threshold in unanesthetized humans. AB - BACKGROUND AND PURPOSE: Hand warming and face warming, combined with inhalation of heated air, are reported to suppress shivering. However, hand or face temperature contributes only a few percent to control of shivering. Thus, it seems unlikely that manipulating hand or facial skin temperature alone would be sufficient to permit induction of therapeutic hypothermia. We tested the hypothesis that focal arm (forearm and hand) warming or lower facial warming, combined with inhalation of heated and humidified gas, only minimally reduces the shivering threshold (triggering core temperature). METHODS: We studied 8 healthy male volunteers (18 to 40 years of age) on 3 days: (1) control (no warming), (2) arm warming with forced air at approximately 43 degrees C, and (3) face warming with 21 L/min of air at approximately 42 degrees C at a relative humidity of 100%. Fluid at approximately 4 degrees C was infused via a central venous catheter to decrease tympanic membrane temperature 1 degrees C/h to 2 degrees C/h; mean skin temperature was maintained at 31 degrees C. A sustained increase in oxygen consumption quantified the shivering threshold. RESULTS: Shivering thresholds did not differ significantly between the control (36.7+/-0.1 degrees C), arm-warming (36.5+/-0.3 degrees C), or face-warming (36.5+/-0.3 degrees C; analysis of variance, P=0.34) day. The study was powered to have a 95% probability of detecting a difference of 0.5+/-0.5 degrees C (mean+/-SD) between control and either of the 2 treatments at alpha=0.05. CONCLUSIONS: Focal arm or face warming did not substantially reduce the shivering threshold in unanesthetized volunteers. It thus seems unlikely that these nonpharmacological modalities will substantially facilitate induction of therapeutic hypothermia. PMID- 12775890 TI - Triggers of subarachnoid hemorrhage: role of physical exertion, smoking, and alcohol in the Australasian Cooperative Research on Subarachnoid Hemorrhage Study (ACROSS). AB - BACKGROUND AND PURPOSE: Unaccustomed strenuous physical exertion can trigger myocardial infarction, but little is known about the mechanisms precipitating subarachnoid hemorrhage (SAH). METHODS: We identified all cases of first-ever SAH among the combined populations (2.8 million) of 4 urban centers in Australia and New Zealand. Information on the type, time, and intensity of exposures in the 26 hours before the onset of SAH was ascertained by structured interviews. We used the case-crossover technique to assess the risk of SAH associated with transient exposures of moderate to extreme physical exertion, heavy cigarette smoking, and binge alcohol consumption. RESULTS: We registered 432 first-ever cases of SAH (62% women; mean age, 56.5 years). A definite time of onset of SAH was established for 393 patients (91%), and information on the levels of physical activity in the preceding 26 hours was obtained in 338 (78%). Of these patients, 19% engaged in moderate to extreme exertion (>or=5 metabolic equivalents) in the 2 hours before SAH, which was associated with a tripling in the risk of SAH (odds ratio [OR], 2.7; 95% CI, 1.6 to 4.6). There was no evidence of any association between heavy cigarette smoking or binge drinking and risk of SAH in the subsequent 2 hours (OR, 1.1; 95% CI, 0.4 to 3.7; and OR, 0.41; 95% CI, - infinity to 5.3). Habitual exercise did not appear to alter the risk of SAH associated with moderate to extreme exertion. CONCLUSIONS: Moderate to extreme physical exertion tripled the risk of SAH, but there was no association between transient heavy smoking or binge drinking and risk of SAH. These data suggest that heavy physical activity may trigger SAH. PMID- 12775891 TI - Human monoclonal IgM antibodies with apoptotic activity isolated from cancer patients. AB - Monoclonal antibodies are accepted as ideal adjuvant therapeutic reagents for all kinds of diseases. Polyvalent (cross-linking) and low-mutated IgM antibodies (less immunogenic) are believed to be the most effective weapons against cancer. The best sources for these types of antibodies are the cancer patients themselves. Using conventional hybridoma technology, not only are fully human monoclonal IgM antibodies isolated, but also new tumor-related targets can be identified using the same experimental approach. The resulting antibodies can be used directly for therapeutic purposes without further modulation and manipulation. This report describes five newly established human monoclonal IgM antibodies; antibody LM-1 that was isolated from a patient with lung cancer, antibodies PM-1 und PM-2 that were isolated from a patient with pancreatic cancer, and antibodies CM-1 and CM-2 which were isolated from a patient with colon carcinoma. The mainly germ-line encoded antibodies are specific for malignant tissues and show only restricted reactivity with healthy cells. When tested for in vitro functional activity, all five antibodies inhibit tumor cell proliferation of carcinoma cells by inducing apoptosis. PMID- 12775892 TI - Human single-chain Fv (scFv) antibody specific to human IL-6 with the inhibitory activity on IL-6-signaling. AB - Human anti-IL-6 antibody may be useful for the immunotherapy of various inflammatory diseases, such as rheumatoid arthritis. Since IL-6 is a growth factor for B cell hybridoma, it is not easy to isolate murine B cell hybridomas producing the anti-IL-6 antibody with the IL-6-signaling inhibitory activity. In this study, the antibody library (Vgamma-Vkappa, Vgamma-Vlambda, Vmu-Vkappa or micro-Vlambda ligated into the pCANTAB 5E phagemid vector) was prepared from peripheral blood mononuclear cells of 20 healthy subjects. The phage display library was panned with an IL-6-coated plastic plate, and the binding specificity was confirmed by ELISA and BIAcore. From the antibody library (Vgamma-Vlambda), five IL-6-specific phage clones were isolated. The effects of the soluble scFvs purified from these phage clones were tested on the growth of the IL-6-dependent human cell line, KT-3. Two of these clones significantly inhibited the growth of KT-3, and three showed no inhibition. PMID- 12775893 TI - Nature's best weapons to fight cancer. Revival of human monoclonal IgM antibodies. AB - The unique features of monoclonal antibodies (specificity, effectiveness, purity and unlimited reproducibility) make them ideal tools for the specific treatment of all kind of diseases. The third generation of monoclonal antibodies for the treatment of human diseases will be, after murine and "humanised" murine immunoglobulins, fully human antibodies. The best source of human monoclonal antibodies are the antibody pools of cancer patients themselves with the best technique for generating them being conventional human hybridoma technology. This technique, will generate human monoclonal antibodies which will not only define important new targets on cancerous tissue, but will also provide the necessary therapeutic human antibodies in the fight against cancer. PMID- 12775895 TI - Is whole body impedance a predictor of blood viscosity? AB - Bioelectrical impedancemetry (BIA) has received a widespread interest as a non invasive approach to body fluid volumes. Since similar techniques have been studied to assess in vitro rheological properties of blood, we investigated the relationships between whole body impedance and blood viscosity parameters in order to determine possible predictive equations. 30 sportsmen (24.6+/-1.01 years; 73.96+/-1.62 kg; 177.73+/-1.33 cm) were enrolled into the study. Body composition was assessed with a multifrequency bioelectrical impedancemeter (Dietosystem Human IM Scan) using low intensity at the following frequencies: 1, 5, 10, 50 and 100 kHz. Viscometric measurements were done at 1000 s(-1) with a falling ball viscometer (MT 90 Medicatest). Hematocrit (Hct) was measured with microcentrifuge. A standardized exercise test was performed on a cycloergometer during 25 minutes. Physical working capacity (W170) was calculated and VO2max was evaluated with Astrand nomograms. Two hemorheological parameters were independently correlated with impedance (Z) measurements: whole blood viscosity (WBV) at 100 kHz (r=0.518; p=0.01) and Hct at 1 kHz (r=-0.485; p=0.01). Plasma viscosity was correlated multilinearly with water/fat free mass and Z at 10 kHz (r=0.441; p=0.02). In addition both WBV and Z at 100 kHz exhibited correlations with aerobic working capacity (VO2max ) with r=-0.482 and r=-0.475 (p or = 0.6 to < or = 1.4 in association with Z values from -2 to 2. The five misfit items have to be revised. The study has indicated that DOA is a valid instrument in psychiatric work rehabilitation and ought to be useful in the dialogue between occupational therapists and clients about goalsetting and treatment planning. PMID- 12775928 TI - Workplace assessments and functional capacity evaluations: current beliefs of therapists in Australia. AB - OBJECTIVES: The purpose of this study was to understand the current beliefs of therapists in Australia, and the strategies they use to address the issues of credibility, reliability, consistency, trustworthiness, validity, generalisability and quality in conducting work-related assessments. STUDY DESIGN: In-depth semi-structured interviews were conducted with 26 occupational therapists and physiotherapists from around Australia. FINDINGS: Participants expressed the belief that the therapist was the assessment instrument and was central to the credibility of an assessment. Conflict was reported when participants modified standardised assessments in an attempt to focus on context relevant activities and tasks. Participants were aware of the issues of reliability and validity but believed it was not practical to establish these aspects formally in most work-related assessments. The strategies used to achieve credibility, reliability, consistency, trustworthiness, validity, generalisability and quality were similar to those recommended for use in qualitative research. CONCLUSIONS: The strategies identified in this study can provide the basis for therapists to examine how they conduct work-related assessments and consider whether they currently use these strategies or have the opportunity to implement others. PMID- 12775929 TI - The effects of a continuous passive motion device on myoelectric activity of the erector spinae during prolonged sitting at a computer workstation. AB - Continuous passive motion (CPM) has been proposed as a method to help individuals with low back pain cope with prolonged sitting. The purpose of this study was to investigate the effects that a commercially available CPM device had on lumbar and thoracic erector spinae (ES) muscle activation (using surface electromyography, "EMG"), and on subjective discomfort during prolonged seated computer work with and without the use of the CPM device. There were no significant differences in average ES muscle activation levels, amplitude probability distribution functions, and EMG gaps number and length when sitting with the CPM device was compared to sitting normally. Subjective ratings of discomfort were also not significantly different between the two sitting conditions. The results indicated that there were no clear mechanisms by which the CPM device may reduce ES muscle pain and fatigue for the tasks and pain-free individuals studied. PMID- 12775930 TI - Experience of cumulative trauma disorders on life roles of worker and family member: a case study of a married couple. AB - The prevalence of diagnosed cumulative trauma disorders (CTD) within the workforce comes at a high price for employers burdened with financial losses from missed work and worker's compensation costs. Research has focused primarily on the impact of CTD on the worker role within the workplace, overlooking the impact on roles across multiple environments [24,35,54]. Furthermore, the influence of CTD on life roles of a spouse has not been examined. This single case study illustrated the experience of CTD within a marital relationship through the use of grounded theory. Results indicated that adaptations to CTD symptoms were least altering to the established routines and roles of the couple. With progression of symptoms, the spouse without symptoms was relied on more heavily for adaptations to manage pain. The results of this study indicate that occupational therapists must examine the client's valued roles and incorporate the family into intervention strategies. PMID- 12775931 TI - The effectiveness of back pain and injury prevention programs in the workplace. AB - Musculoskeletal disorders in the workplace cause thousands of injuries and cost industry billions of dollars yearly. Work injury prevention programs have been developed and implemented as a means for cost containment. A variety of preventive strategies have been investigated in primary research. The purpose of this review article is to examine the effectiveness of back injury and pain prevention programs in the workplace. Nine studies published between 1995 and 2000 were reviewed and analyzed. Studies used primarily one of three types of preventive strategies: 1) back belts, 2) education and task modification, and 3) education and task modification with workstation redesign. The effectiveness of back belts to prevent back pain and injury remains inconclusive. Positive outcomes were associated with studies reporting high compliance that used job specific and individualized/small group education and training approaches. Themes that arose following a critical review of primary research studies are discussed. PMID- 12775932 TI - The impact of mental illness on employment: consumers' perspectives. AB - This paper is based on a qualitative study of employment for people with mental illness. Forty-one consumers of mental health services were interviewed to obtain their perspectives on employment. One of the major issues they discussed was the ways in which mental illness affected their employment experiences. These effects were complex and interrelated and varied between individuals depending on their unique characteristics and circumstances. Participants described effects relating to the need to maintain mental health, difficulties with work performance, and work confidence and work goals. Understanding consumers' perspectives on how mental illness affects employment is necessary for practitioners and researchers seeking to explain or improve consumers' employment outcomes. PMID- 12775933 TI - Interaction of laminar airflow with viscoelastic airway mucus. AB - Basic wave interaction mechanism between the laminar airflow and viscoelastic layer in a rigid tube is investigated numerically. The purpose is to explore the effect of mucus viscoelasticity on the stability of the coupled airflow-mucus system in pulmonary airways under clinical conditions where the serous layer is absent. The results indicate that the onset flow speed, for the initiation of unstable surface waves, is very sensitive to mucus viscosity and it may be as high as 35 times the elastic case for a very viscous mucus with the same elasticity. While the onset speed and wavelength increases, wave speed decreases with increasing mucus viscosity, reducing from about 40% of the flow speed for elastic mucus to less than 1% for a very viscous mucus. Also, a case study for a patient with chronic bronchitis shows that large amplitude waves may form on the mucus surface during forced expiration. PMID- 12775934 TI - Biomechanics of the knee joint in deep flexion: a prelude to a total knee replacement that allows for maximum flexion. AB - A two-dimensional anatomically based mathematical model of the human knee joint was developed to understand its biomechanics in deep flexion. The model was used to determine the internal knee loads as it simulates isometric quadriceps and hamstring co-contractions at different flexion angles during deep squat. It was found that in order to achieve deep flexion, large muscle forces are required, resulting in large tibio-femoral contact forces. In deep flexion, the femoral contact point was located on the most proximal point of the posterior condyle, location which was not affected by the level of quad activation. Conversely, the location of the tibial contact point was highly affected by the level of quad activation. Both anterior and posterior fiber bundles of the posterior cruciate ligament were found to carry high loads when the knee is maximally flexed. These results point to the important role of the posterior cruciate ligament in this position, and suggest the necessity of retaining this ligament during total knee replacement (TKR) procedures that allows for maximum flexion angles. Furthermore, the present data provide an explanation why most TKR's do not allow deep flexion: while contact occurs on the most proximal points of the posterior condyles in normal knees, this portion of the condyles is not presently resurfaced when performing a TKR. PMID- 12775935 TI - Subject-specific compressive tolerance estimates. AB - Spinal load models have become an increasingly valuable tool for the evaluation of the stress placed on the spine. In order to get an accurate representation of these spinal loads, they must be compared to known tolerance values. Bone mineral content and density of the lumbar spine of 23 males and 21 females was measured using a dual energy x-ray absorptiometry. Compression tolerance values were predicted by previously published studies based upon bone mineral levels. Anthropometric measurements were recorded and related to the compression tolerance values through the use of multivariate linear regression techniques. Compression tolerance values based on the bone mineral content or density explains more of the individual variability than solely age-based estimates. Several anthropometric variable regression models were developed that resulted in moderate to good predictive power (R(2) = 0.62 to 0.81). The current study describes a useful alternative to traditional tolerance estimates that accounts for individual differences requiring non-invasive and time-efficient procedures. PMID- 12775936 TI - Home informatics in healthcare: assessment guidelines to keep up quality of care and avoid adverse effects. AB - Due to an ageing population and improved treatment possibilities, a shortage in hospital beds is a fact in many countries. Home healthcare schemes using information technology (IT) are under development as a response to this and with the intention to produce a more cost-effective care. So far it has been shown that home healthcare is beneficial to certain patient groups. The trend is a widening of the criteria for admission to home healthcare, which means treatment in the home of more severe conditions that otherwise would require in-hospital care. Home informatics has the potential to become a means of providing good care at home. In this process, it is important to consider what new risks will be encountered when placing electronic equipment in the home care environment. Continuous assessment and guidance is important in order to achieve a safe and effective care. Based on a review of current knowledge this paper presents an inventory of risks and adverse events specific to this area. It was found that risks and adverse events could stem from technology in itself, from human technology interaction conditions or from the environment in which the technology is placed. As a result from the risk inventory, this paper proposes guidelines for the planning and assessment of IT-based hospital-at-home schemes. These assessment guidelines are specifically aimed at performance improvement and thus to be considered a complement to the more general guidelines on telehomecare adopted by the American Telemedicine Association (ATA) in October 2002. PMID- 12775937 TI - Low-cost antibiotic loaded systems for developing countries. AB - Treatment of osteomyelitis with local antibiotic delivery systems has become a common practice in orthopaedic surgery. This study attempted to show that locally produced pure or bioglass reinforced plaster of Paris, hydroxyapatite and sodium alginate are promising biomaterials and mainly because of economical reasons and availability, may be an alternative in clinical practice, especially for developing countries. A total of 32 rabbits were divided into four groups (n:8). In group A, sodium alginate + cephazoline; in group B, plaster of Paris + bioglass + cephazoline; in group C, plaster of Paris + hydroxyapatite + cephazoline and in group D, plaster of Paris + cephazoline were used. The blood serum cephazoline concentrations were analyzed by high performance liquid chromatography on days 1 to 10 everyday and then at days 13, 17, 18, 24, 25 and 30. The mean values +/- standard deviations and median values of blood serum antibiotic concentrations for groups A, B, C and D were 1.45 +/- 0.40 (1.42) mcg/ml, 1.53 +/- 0.64 (1.31) mcg/ml, 1.92 +/- 0.39 mcg/ml (1.90) and 1.41 +/- 0.65 (1.25) mcg/ml, respectively. The detected antibiotic level was constantly over the minimum inhibitory concentration for Staphylococcus aureus. In conclusion, it can be stated that these materials are promising as a antibiotic delivery system even with simple production methods. PMID- 12775938 TI - Chk'n out in mitosis. PMID- 12775939 TI - Phosphorylation of Xenopus Cdc25C at Ser285 interferes with ability to activate a DNA damage replication checkpoint in pre-midblastula embryos. AB - We have recently demonstrated that negative regulation of human Cdc25 protein phosphatases by phosphorylation at their 14-3-3 site can be antagonized through phosphorylation at an adjacent site in the -2 position.1 Based on structural homology for different Cdc25 phosphatases, a similar regulatory pathway also could be conserved in Xenopus embryos, where cell cycle checkpoints are not operational prior to the Midblastula Transition (MBT). Here, we demonstrate that before MBT, XeCdc25C is phosphorylated on Ser285, an analogous site to Ser214 in human Cdc25C or Ser307 Cdc25B.(1) Phosphorylation of Ser285 prevents subsequent inhibitory phosphorylation of XeCdc25C on Ser287, thus maintaining XeCdc25C in an active form. Mutation of Ser285 to alanine allows the reconstitution of a DNA damage replication checkpoint. This effect is completely dependent on Ser287 phosphorylation as additional mutation of Ser287 to alanine fully reversed the cell cycle inhibitory effect of Ser285A XeCdc25C. We propose that phosphorylation of XeCdc25C Ser285 may account for the lack of a DNA replication checkpoint in cleaving Xenopus embryos prior to the MBT. PMID- 12775942 TI - New-onset diabetes after transplantation: 2003 International consensus guidelines. Proceedings of an international expert panel meeting. Barcelona, Spain, 19 February 2003. PMID- 12775943 TI - To err is human: to err less is mandatory. PMID- 12775944 TI - Atypical glandular cells: new Bethesda Terminology and Management Guidelines. AB - Although Pap tests have enabled early detection of premalignant lesions, the introduction of new collecting devices has significantly improved the detection of lesions hidden in the endocervical canal, such as adenocarcinoma in situ (AIS). The term "atypical glandular cells of undetermined significance" (AGUS) was introduced at the 1988 Bethesda Conference and defined as morphologic changes in glandular cells beyond those that are suggestive of the benign reactive process, but insufficient for the diagnosis of adenocarcinoma in situ (AIS). In the new 2001 Bethesda System, the term has been eliminated and replaced with the term "atypical glandular cells" (AGC), with the following subclassifications: not otherwise specified (NOS), favor neoplasia, endocervical AIS, and adenocarcinoma. The risks of premalignant or malignant disease associated with the AGC favor neoplasia category are substantially higher than in the AGC NOS category (96% vs. 9-41%, respectively). Patients diagnosed with AGC NOS or AGC favor neoplasia will require colposcopy, endocervical sampling, and, for patients over 35 years of age, endometrial biopsy. If all of these tests are negative, the Pap test should be repeated in 4-6 month intervals until 4 consecutive normal tests are obtained. Positive results in one of the tests will require management according to ASCCP guidelines. The AGC favor neoplasia diagnosis also requires cervical conization and/or other testing, as the incidence of premalignant or malignant lesions in patients with this diagnosis is high. The role of HPV testing in this setting is unknown at this time. PMID- 12775945 TI - Mild fetal cerebral ventriculomegaly: diagnosis, clinical associations, and outcomes. AB - The normal fetal lateral ventricular diameter remains stable at 10 mm over gestation. Mild ventriculomegaly, defined as a lateral ventricular diameter of >or=10 mm but or=3 mm but /=140 mmHg, DBP>/=90 mmHg, or both or prescription of antihypertensive medication) compared with the presence of no risk factors was 1.41 (95% CI, 1.21 to 1.64), 1.64 (95% CI, 1.38 to 1.96), 1.93 (95% CI, 1.56 to 2.39), 2.01 (95% CI, 1.51 to 2.68) and 3.34 (95% CI, 2.07 to 5.38) (P value for trend < 0.001) for respective risk factors of 1, 2, 3, 4 and >/=5. Even after the subjects were stratified according to blood pressure, the clustering of risk factors was associated with an increased risk of hypertension for subjects in all three categories of normotension: low-normal, normal, and high-normal. CONCLUSIONS: Clustering of cardiovascular risk factors associated with hypertension precedes an increase in the risk of hypertension in Japanese men. PMID- 12775956 TI - Alcohol use and cardiovascular disease preventive services. AB - BACKGROUND: Moderate drinking is associated with decreased cardiovascular mortality. Biological effects may mediate this association, but differences in utilization of preventive services may be important. DESIGN AND METHODS: Cross sectional analysis of adults participating in the Third National Health and Nutrition Examination Survey with data on alcohol use and prior use of services for the detection and treatment of hypertension and hypercholesterolaemia. Regression models were analysed to provide age-, sex- and race-adjusted risk estimates for outcomes related to these services for several patterns of alcohol use relative to regular light drinkers. RESULTS: Hypertension was more common in most alcohol use strata compared with regular light drinkers (adjusted prevalence ratios 1.22 for lifelong abstainers, 1.33 for nonlifelong abstainers, 1.35 for infrequent moderate drinkers, 2.01 for frequent moderate drinkers, 1.73 for infrequent heavy drinkers and 1.98 for regular heavy drinkers, P-values < 0.05). Having had blood pressure measured was similar in all drinking strata. The prevalence of hypercholesterolaemia was similar in drinking strata, but most drinking patterns were associated with lower use of services to detect high cholesterol (prevalence ratios 0.7 for lifelong abstainers, 0.8 for nonlifelong abstainers, infrequent moderate and regular moderate drinkers, 0.6 for infrequent heavy drinkers, and 0.7 for regular heavy drinkers, P-values < 0.05). CONCLUSIONS: The utilization of cardiovascular disease preventive services may contribute to the beneficial association of moderate drinking with cardiovascular mortality. Additional research is needed to estimate the influence of differences in health services utilization on the association of alcohol use with cardiovascular disease outcomes. PMID- 12775957 TI - Hair loss, insulin resistance, and heredity in middle-aged women. A population based study. AB - CONTEXT: The association of androgenic alopecia (AGA) with insulin resistance, coronary artery disease and hypercholesterolemia has been previously reported in men, but no such association has been reported in women with female androgenic alopecia (AGA). Female AGA has usually been linked with hyper-androgenism and hirsutism and, most recently, also with polycystic ovarian syndrome (PCOS), even though epidemiological documentation of the latter association is scanty. Polycystic ovarian syndrome is quite common among Caucasian women, and its association with insulin resistance is well documented. OBJECTIVES AND DESIGN: The aim of this study was to obtain a more precise estimation of the prevalence on female AGA and to describe its possible connections with insulin resistance linked parameters and with paternal and maternal family history of alopecia. A cross-sectional population based cohort survey was carried out in the City of Oulu, Finland in 1998. SETTING AND PARTICIPANTS: As a part of a population based cohort study the hair status of 324 women aged 63 years was assessed by a modification of Ludwig's scale. The background data consisting of anthropometric measures (weight, height, body mass index, waist, hip and neck circumferences), smoking status, chronic diseases and their medication as well as the family history of AGA were collected by questionnaires and interviews made by study nurses and in clinical examination. Blood samples for laboratory tests were taken on the same occasion. RESULTS: The prevalence of extensive loss of hair (at least grade II or III on Ludwig's scale) was quite high (31.2%). The insulin resistance associated parameters, such as waist and neck circumferences, abdominal obesity measured by waist-to-hip ratio, mean insulin concentration (11.3 mU/l versus 9.95 mU/l, p=0.02) or urinary albumin-to-creatinine ratio (1.80 versus 1.58, p=0.01), were significantly higher in women with extensive hair loss compared to those with normal hair or only minimal hair loss (grade I on Ludwig's scale). The women belonging to the highest quintiles of neck or waist circumferences had significantly increased risk for extensive hair loss compared to those with normal hair or minimal hair loss, the unadjusted ORs being 2.25 (95% CI, 1.26 4.03) and 1.75 (95% CI, 1.00-3.07), respectively. Similarly in women with hyperinsulinemia (fs-insulin >10 mU/l), microalbuminuria (urinary albumin-to creatinine ratio exceeding the highest microalbuminuria decile (>2.5 mg/mmol) and paternal history of AGA the ORs for alopecia were increased being 1.65 (95% CI, 1.02-2.67), 2.39 (95% CI, 1.21-4.73) and 2.08 (95% CI, 1.26-3.44). All of these ORs, except those for highest quintiles of waist and neck circumferences remained significant in multiple adjusted models. CONCLUSIONS: According to the results of this study, female AGA (grade II or III on Ludwig's scale) was quite common among Finnish women aged 63 years. Our results support the hypothesis that women with some markers of insulin resistance have significantly increased risk for female AGA. Paternal history of alopecia seemed to be more common in female AGA compared to women with normal or minimal loss of hair. PMID- 12775958 TI - Stent-induced restenosis in the swine coronary artery is inhibited by a platelet derived growth factor receptor tyrosine kinase inhibitor, TKI963. AB - Activities of vascular smooth muscle cells (SMCs) such as proliferation, migration, and matrix production contribute to restenosis following clinical interventions of angioplasty and stent placement. Because activation of platelet derived growth factor (PDGF)-receptor tyrosine kinase (PDGFr-TK) influences these processes and promotes restenosis, TKI963, an inhibitor of the PDGFr-TK was discovered, and its efficacy was evaluated in blocking stent-induced restenosis as analyzed by intravascular ultrasound (IVUS). TKI963, a low-molecular-weight compound, inhibited the cell-free PDGFbetar-TK with a K(i) value of 56 +/- 14 nM. TKI963 also inhibited PDGF-dependent events in human aortic SMCs (e.g., in situ PDGFr autophosphorylation, mitogenesis, chemotaxis, and collagen production with median inhibitory concentration values of approximately 300 nM) without affecting the activity of a series of membrane receptor tyrosine kinases and intracellular serine/threonine kinases. In vivo, stent-induced restenosis in the swine coronary artery was reduced by oral administration of TKI963 (1.25, 2.5, and 5 mg/kg BID, for 28 days). Late lumen cross-sectional area (CSA) loss, plaque CSA growth, and plaque volume in the stent determined by IVUS were dose-relatedly decreased (33 62% at 1.25 mg/kg BID to 66-92% at 5 mg/kg BID, depending on the parameter) compared with controls. TKI963 treatment of 220 bpm. At all tested cycle lengths, haloperidol caused a significant lengthening of Q(T) intervals, which was inversely dependent on heart rate. Numeric analysis suggested that the excessive increase in conduction slowing at rates >220 bpm was due to the drug's Q(T)-prolonging effect, indicating that, at short cycle lengths, the impulses encroached on the refractory period. Thus, in vivo, haloperidol slows intracardiac conduction with rapid on/off kinetics, comparable to the class I antiarrhythmic agent lidocaine. The Q(T) prolongation by haloperidol may lead to an excessive conduction slowing at high heart rates. PMID- 12775965 TI - Effects of ACE inhibition on myocardial apoptosis in an ischemia-reperfusion rat heart model. AB - Myocardial ischemia-reperfusion injury involves necrosis and apoptosis. The inhibition of angiotensin-converting enzyme (ACE) has been reported to suppress infarct size. In this study, it was investigated whether an ACE inhibitor affected myocardial apoptosis and apoptosis-related proteins in rats with experimental myocardial infarction. Anesthetized Sprague-Dawley rats were divided into four groups. Group I underwent 30 minutes of left coronary artery occlusion followed by 24 hours of reperfusion (control group); Group II underwent oral administration of the ACE inhibitor quinapril (10 mg/kg/day) before coronary occlusion (quinapril group); Group III underwent administration of the bradykinin B(2)-receptor antagonist Hoe 140 (250 microg/kg/day, subcutaneously) with quinapril (quinapril + Hoe 140 group); and Group IV underwent administration of Hoe 140 alone (Hoe 140 group). After reperfusion, myocardial infarct size was determined by triphenyltetrazolium chloride staining. Myocardial apoptosis was detected immunohistologically using terminal deoxynucleotidyl transferase mediated nick end labeling staining and DNA electrophoresis. Myocardial caspase-3 activation was analyzed by Western blot and the expressions of Bcl-xL and Bax proteins were detected immunohistochemically. Quinapril significantly reduced the ratio of myocardial infarct size in the ischemic area at risk. In addition, quinapril significantly suppressed the incidence of apoptotic myocytes around the necrotic region (from 18.9 +/- 0.8% to 8.6 +/- 1.0%; P < 0.0001), the intensity of DNA ladder formation, and the activation of caspase-3. Hoe 140 attenuated these protective effects of quinapril. In the immunohistochemical study, Bax and Bcl-xL were expressed in myocytes, and ischemia-reperfusion abolished both proteins in the center region of ischemia. The Bax staining was equally observed among all groups. However, Bcl-xL staining remained in the ischemic area widely after quinapril treatment. In addition, Hoe 140 also depleted this effect of quinapril. These results suggest that inhibition of ACE reduces myocardial infarction and apoptosis via the bradykinin B(2) receptor in part. The antiapoptotic effect of the ACE inhibitor is attributed to the changing expression of Bcl-xL. PMID- 12775966 TI - Effects of the endothelin a receptor antagonist darusentan on blood pressure and vascular contractility in type 2 diabetic Goto-Kakizaki rats. AB - The present study evaluated the effects of long-term treatment with the endothelin A (ET(A)) receptor antagonist darusentan (LU135252) on blood pressure (BP) and vascular target-organ damage in spontaneously type 2 diabetic Goto Kakizaki (GK) rats. BP was monitored by radiotelemetry in untreated and darusentan-treated GK rats from 10-24 weeks of age. Relaxation of mesenteric artery segments by acetylcholine (ACh) and sodium nitroprusside (SNP) was measured to assess endothelium-dependent and -independent vasorelaxation. Aortic soluble guanylyl cyclase (sGC) activity was studied in vitro after stimulation by the nitric oxide (NO) donor diethylamine-NONOate. Untreated GKs were mildly hypertensive and showed a blunted vascular relaxation by ACh and SNP and a reduction in NO-stimulated sGC activity in comparison with Wistar control rats. Darusentan led to a small but sustained reduction in 24-h BP but did not restore the endothelium-dependent vasorelaxation nor the NO-stimulated cGMP formation in GK rats. The present findings suggest that an activated endothelin pathway may contribute to elevated BP but is not involved in vascular dysfunction in this animal model of type II diabetes. PMID- 12775967 TI - Deficiency of nitric oxide synthase 2 results in increased neointima formation in a mouse model of vascular injury. AB - Restenosis frequently occurs after arterial interventions. The inducible form of nitric oxide synthase (NOS2) may both promote and inhibit neointima formation. This study investigated the role of NOS2 for neointima formation in a mouse model of carotid artery injury. The common carotid artery was ligated in anesthetized mice. Homozygous NOS2 knockout mice were compared with wild-type B6/129 mice or wild-type mice treated with the pharmacologic NOS2 inhibitor aminoguanidine given orally daily after ligation (n = 6-8 in each group). Vessels were harvested for quantification of lesion size 4 weeks later, or serially after ligation for tissue analysis. mRNA for NOS2 increased 1-4 days after ligation of the carotid artery. Cell proliferation could be visualized with an antibody against proliferating cell nuclear antigen. An intimal smooth muscle cell layer, confirmed by an alpha-actin antibody, was observed in the lumen 4 weeks after injury. Inhibition of NOS2 by either pharmacologic or genetic approaches tended to increase the area of intima formation (P = 0.13 or P < 0.05, respectively) and increased the intima/media ratio (P = 0.14 and P < 0.01, respectively). Inhibition of NOS2 by two different approaches increased neointima formation in a mouse model of mechanical vessel injury, indicating that the NOS2 expressed in the injured vessel wall is beneficial. PMID- 12775968 TI - Intracellular alkalinization augments capacitative Ca2+ entry in vascular smooth muscle cells. AB - Agonist-induced Ca2+ influx of vascular smooth muscle cells is thought to be triggered by depletion of intracellular Ca2+ stores. This study investigated the effects of intracellular alkalinization on capacitative Ca2+ entry in A7r5 rat aortic smooth muscle cells. Intracellular alkalinization was induced by NH(4)Cl. Transplasmalemmal Ca2+ influx due to Ca2+ store depletion induced by thapsigargin, which was abolished by pretreatment of the cells with SKF-96365 but not affected by that with verapamil, was significantly increased by pretreatment with NH(4)Cl. Neither 5-hydroxytryptamine-induced inositol monophosphate accumulation nor intracellular Ca2+ release from its stores was affected by NH(4)Cl. These results suggest that intracellular alkalinization acts on the process(es) after depletion of Ca2+ stores and facilitates capacitative Ca2+ entry in vascular smooth muscle cells. PMID- 12775969 TI - Effect of continuous subcutaneous treprostinil therapy on the pharmacodynamics and pharmacokinetics of warfarin. AB - Treprostinil sodium was recently approved in the United States for continuous subcutaneous infusion in the treatment of pulmonary arterial hypertension (PAH). Anticoagulation with warfarin is recommended in PAH therapy. Given the likelihood for treprostinil and warfarin coadministration, a single-blind, controlled, crossover study was conducted to evaluate the effect of treprostinil infusion on the pharmacodynamics and pharmacokinetics of a single dose of warfarin. Area under the effect-time curve (AUEC(0-1)) and maximum effect over the entire sampling phase (E(max)) for warfarin INR were 219.58 and 2.071 with treprostinil and 218.93 and 2.041 with vehicle, respectively. Mean time to attain the peak concentration of R-enantiomer of warfarin (T(max)), half-life, and elimination rate constant (k(el)) were 1.9 hours, 51.688 hours, and 0.0137 per hour, respectively, in the presence of treprostinil and 1.5 hours, 52.579 hours, and 0.0137 per hour, respectively, in the presence of vehicle (control). Results were similar for the S-enantiomer. The 90% confidence intervals for warfarin INR and warfarin R- and S-enantiomer pharmacokinetic parameter (C(max) and AUC( infinity )) ratios were within 0.80-1.25, which was established as the no-effect criterion for treprostinil coadministration. No serious or severe adverse events, anticoagulation-related events, or clinically significant physical or laboratory findings were reported. These findings suggest that a clinically important interaction between treprostinil and warfarin during therapy is unlikely. PMID- 12775970 TI - Defibrillatory action of glibenclamide is independent from ATP-sensitive K+ channels and free radicals. AB - This study investigated whether glibenclamide exerts a defibrillatory action and if this action is mediated by a blockade of ATP-sensitive K+ channels (K(ATP)) or by an anti-free radical mechanism. Aerobically perfused isolated rat hearts were subjected to 10 min of pacing-induced ventricular fibrillation (VF) followed by 10 min of perfusion without pacing (post-VF period), in the presence of solvent (controls), 1 microM K(ATP) blocker glibenclamide, 10 microM K(ATP) opener cromakalim, and their combination, respectively. In controls, pacing-induced VF caused a significant deterioration in cardiac function in the post-VF period. Spontaneous defibrillation was 42%. Glibenclamide improved post-VF cardiac function and resulted in 100% (P < 0.05) spontaneous defibrillation. Cromakalim did not significantly affect post-VF cardiac function and the incidence of spontaneous defibrillation as compared with controls. The combination of the compounds improved cardiac function and resulted in 83% (P < 0.05) spontaneous defibrillation. In separate experiments, 2,5-dihydroxybenzoic acid formation in the perfusate as a marker of hydroxyl radical formation was measured by high performance liquid chromatography and cardiac superoxide production was assessed by lucigenin-enhanced chemiluminescence during pacing-induced VF. Glibenclamide did not affect hydroxyl radical generation or myocardial superoxide content during VF. The conclusion is that glibenclamide exerts a defibrillatory action and improves post-VF cardiac function in rat hearts and these effects are independent from K(ATP) and free radicals. PMID- 12775971 TI - Efficacy of edaravone, a free radical scavenger, on left ventricular function and structure in diabetes mellitus. AB - This study was designed to assess the efficacy of edaravone (3-methyl-1-phenyl-2 pyrazolin-5-one), a free radical scavenger that possesses anti-oxidant effects, on cardiac function and fine structure of the left ventricular myocardium in diabetes mellitus. Male Otsuka Long-Evans Tokushima Fatty (OLETF) rats, a model of spontaneous development of type II diabetes (30 weeks; n = 15) were divided into two groups and treated with edaravone 30 mg/kg/d or vehicle for 2 weeks. OLETF rats showed hyperglycemia (352 +/- 71 mg/dl vs normal control; 128 +/- 52 mg/dl), increased thiobarbituric acid-reactive substances (TBARS; 6.9 +/- 2.5 nM/ml vs 2.8 +/- 0.6 nM/ml), and decreased superoxide dismutase activity (21.5 +/ 0.9 U/ml vs 25.8 +/- 0.7 U/ml). Increased left ventricular end-diastolic pressure (12 +/- 3 mm Hg vs 6 +/- 2 mm Hg) and hypertrophied cardiocytes (23.1 +/ 1.4 vs 17.6 +/- 1.0 microm) were also observed (P < 0.05, respectively). Edaravone could not improve plasma glucose level and hemodynamic parameters but significantly decreased TBARS values (3.8 +/- 0.5) and increased superoxide dismutase activity (24.5 +/- 0.8) (vs OLETF, P < 0.05, respectively). Moreover, edaravone effectively preserved cardiocyte diameter (18.2 +/- 0.9 microm) and the fine structure of mitochondria. Thus, edaravone exhibits modest cardiac protection in diabetes mellitus independent of blood sugar level. PMID- 12775972 TI - Constitutional rho-kinase regulates atrioventricular nodal conduction and ventricular repolarization of the canine heart. AB - Given the limited information, physiological roles of Rho-kinase in the cardiac conduction system and ventricular repolarization process were assessed in comparison with those in the coronary vascular tone. A specific Rho-kinase inhibitor Y-27632 was administered to the nutrient coronary artery of the canine isolated, blood-perfused atrioventricular node preparation under the monitoring of the ventricular monophasic action potentials. Administration of Y-27632 moderately suppressed the atrioventricular nodal conduction, slightly but significantly accelerated the repolarization process, and potently increased the coronary blood flow, whereas it hardly affected the intraventricular conduction. The estimated concentrations of Y-27632 causing the currently observed effects were enough to inhibit Rho-kinase. These results suggest that constitutional Rho kinase functions to moderately facilitate the atrioventricular nodal conduction, slightly delay ventricular repolarization process, and significantly increase the coronary vascular tone. PMID- 12775973 TI - Risperidone prolongs cardiac repolarization by blocking the rapid component of the delayed rectifier potassium current. AB - Cases of QT prolongation and sudden death have been reported with risperidone, a neuroleptic agent increasingly prescribed worldwide. Although hypokalemia was present in some of these events, we hypothesized that risperidone may have unsuspected electrophysiologic effects predisposing patients to proarrhythmia. In six isolated guinea pig hearts, risperidone elicited prolongation of cardiac repolarization: action potential duration increased from a baseline value of 128 ms +/- 5 to 147 ms +/- 5 (15%) with risperidone 1 microM during pacing at 250-ms cycle length, whereas the increase was only 10%, from 101 ms +/- 2 to 111 ms +/- 4, with pacing at a cycle length of 150 ms. In human ether-a-go-go (HERG) transfected Chinese hamster ovary cells (n = 16), risperidone caused concentration-dependent block of the rapid component (I(Kr)) of the delayed rectifier potassium current with an IC(50) for tail block of 261 nM. Risperidone did not block I(Ks). Risperidone exerts cardiac electrophysiologic effects similar to those of Class III antiarrhythmic drugs. These effects are observed at clinically relevant concentrations. Because risperidone is metabolized primarily by CYP2D6, these actions likely enhance risk for risperidone-related QT prolongation and proarrhythmia in specific patient subsets (e.g., poor metabolizers and those taking interacting drugs). PMID- 12775974 TI - Quinaprilat reduces myocardial infarct size involving nitric oxide production and mitochondrial KATP channel in rabbits. AB - This study examined whether quinaprilat, an angiotensin-converting enzyme inhibitor, reduces the infarct size, and investigated the mechanisms for its infarct size-reducing effect, in rabbits. Japanese white rabbits underwent 30 min of ischemia and 48 h of reperfusion. Quinaprilat (100 microg/kg/h or 300 microg/kg/h for 70 min, IV) was administered 20 min before ischemia with or without pretreatment with Nomega-nitro-l-arginine methyl ester (l-NAME) (10 mg/kg, IV, a nitric oxide synthase inhibitor), 5-hydroxydecanoic acid sodium salt (5-HD) or posttreatment with 5-HD (5 mg/kg, IV, a mitochondrial KATP channel blocker). The area at risk as a percentage of the left ventricle was determined by Evans blue dye and the infarct size was determined as a percent of the area at risk by triphenyl tetrazolium chloride staining. Using a microdialysis technique, myocardial interstitial levels of 2,5-dihydroxybenzoic acid (2,5-DHBA), an indicator of hydroxyl radicals, and NOx, an indicator of nitric oxide, were measured before, during, and after 30 min of ischemia. Quinaprilat significantly reduced the infarct size in a dose-dependent manner (30.1 +/- 3%, n = 10, and 27.6 +/- 2%, n = 7, respectively) compared with the control (46.5 +/- 4%, n = 10). The infarct size-reducing effect of quinaprilat was completely blocked by pretreatment with l-NAME (43.8 +/- 2%, n = 8) and 5-HD (50.1 +/- 3%, n = 8) and posttreatment with 5-HD (50.3 +/- 2%, n = 8), respectively. Quinaprilat did not affect the myocardial interstitial 2,5-DHBA level but significantly increased the NOx level during ischemia and reperfusion. Quinaprilat reduces myocardial infarct size involving NO production and mitochondrial KATP channels in rabbits without collateral circulation. PMID- 12775975 TI - Regulation of protein kinase C isozyme and calcineurin expression in isoproterenol induced cardiac hypertrophy. AB - Protein kinase C (PKC) and calcineurin are known to play a pivotal role in the development of cardiomyocyte growth. However, its role in Isoproterenol-induced (Iso) cardiac hypertrophy has not been characterized so far and were focus of the current study. After chronic beta-adrenergic stimulation of male Wistar rats with Iso (2mg/kg x day) for 2 and 7 days using osmotic minipumps, we determined a) cardiac PKC-activity, b) the expression of cardiac PKC isozymes (PKC-alpha, PKC delta and PKC-epsilon) both at the protein and the mRNA-level and c) the expression of calcineurin using Western blot analysis. Iso-treatment for 2 and 7 days results in cardiac hypertrophy with an increase of the heart weight-to-body weight ratio by 36% and 27%. Iso-induced myocardial growth was associated with an enhanced total PKC-activity and a significant increased protein expression of cytosolic PKC-alpha (day 2: +38%; day 7: +43%), PKC-delta (day 2: 85%; day 7: +78%) and PKC-epsilon (day 7: +58%). The protein amount of calcineurin was not significantly altered by Iso compared with sham-operated controls. The increased expression of PKC-alpha, PKC-delta and PKC-epsilon in the cytosol was paralleled by a transcriptional upregulation of the absolute mRNA-levels of these PKC isozymes as determined by quantitative RT-PCR. PMID- 12775976 TI - Effect of eplerenone, a selective aldosterone blocker, on blood pressure, serum and macrophage oxidative stress, and atherosclerosis in apolipoprotein E deficient mice. AB - Oxidative stress is involved in the pathogenesis of atherosclerosis, and angiotensin II (AT-II) induces oxidative stress and enhances atherogenesis. Aldosterone, which has an important role in the pathology of heart failure, has recently been implicated as a mediator of AT-II biologic activities. In this study, we analyzed whether administration of the selective aldosterone blocker eplerenone to atherosclerotic apolipoprotein E-deficient (E0) mice would affect their oxidative status and atherogenesis. Apolipoprotein E-deficient mice were administered chow containing eplerenone (200 mg/kg/day) for 3 months. Blood pressure, serum and macrophage oxidative status, and aortic atherosclerotic lesion area were evaluated in mice treated with eplerenone compared with untreated mice. Eplerenone administration significantly decreased systolic and diastolic blood pressure by 12% and 11%, respectively, compared with untreated mice. Serum susceptibility to lipid peroxidation decreased by as much as 26%, and serum paraoxonase activity increased by 28% in eplerenone-treated mice compared with untreated mice. Peritoneal macrophages from eplerenone-treated mice contained reduced levels of lipid peroxides, and their macrophage oxidation of low-density lipoprotein (LDL) and superoxide ion release were significantly reduced (by 17% and 43%, respectively), compared to untreated mice. Daily injections of AT-II (0.1 mL, 10(-)7M) during the final 3 weeks of the study in eplerenone-treated mice substantially attenuated the eplerenone-mediated reduction in macrophage superoxide release and LDL oxidation. Finally, the atherosclerotic lesion area in aortas of eplerenone-treated mice was significantly reduced (by 35%) versus untreated mice, and this effect was reversed by AT-II. Administration of the selective aldosterone blocker eplerenone significantly reduced oxidative stress and atherosclerosis progression in E0 mice. These data suggest that aldosterone could have a significant pro-oxidative role in the pathogenesis of atherosclerosis. PMID- 12775977 TI - Eicosapentaenoic acid reduces myocardial injury induced by ischemia and reperfusion in rabbit hearts. AB - Intake of fish oil is known to have cardioprotective effects and reduce cardiovascular mortality. However, it is not widely recognized that eicosapentaenoic acid (EPA), one of the n-3 polyunsaturated fatty acids (PUFAs), exerts beneficial effects against myocardial ischemia/reperfusion injury. The purpose of this study is to investigate whether EPA attenuates the severity of myocardial ischemia/reperfusion injury and which cellular mechanism is involved. Rabbits were treated with or without EPA (600 mg/kg/day) for 2 weeks. Infarct size was measured in open-chest rabbits after 30-minute occlusion of the left anterior descending coronary artery (LAD) and after the subsequent 3-hour reperfusion. In several groups, NG-nitro-L-arginine methyl ester (L-NAME), an inhibitor of nitric oxide (NO) synthase, or charybdotoxin, a blocker of calcium activated potassium (K(Ca)) channels, was infused intravenously beginning 20 minutes before LAD occlusion and continuing during reperfusion. Infarct size was reduced in the group treated with EPA compared with the control group (7.2 +/- 1.0% vs 24.6 +/- 2.3%; P < 0.01). The occurrence of ventricular arrhythmias in the reperfusion period tended to decrease in the EPA group. Either L-NAME or charybdotoxin partially blunted or completely abolished the infarct size-limiting effect of EPA, respectively. Eicosapentaenoic acid significantly increased the n 3:n-6 ratio of PUFA. Eicosapentaenoic acid reduces myocardial infarct size, mainly via the opening of K(Ca) channel-mediated and partially NO-mediated mechanisms in rabbit hearts. PMID- 12775978 TI - Nitric oxide donor molsidomine favors features of atherosclerotic plaque stability during cholesterol lowering in rabbits. AB - Rupture-prone atherosclerotic plaques are characterized by a thin fibrous cap containing numerous macrophage-derived foam cells and few smooth muscle cells (SMC). Decreasing the ratio between macrophages and SMC might favor plaque stabilization. Macrophages expressing inducible nitric oxide (NO) synthase become hypersensitive to killing by exogenous NO donors. Therefore, we investigated in cholesterol-fed rabbits (20 weeks 0.3% cholesterol) the effect of 4 weeks cholesterol withdrawal alone and in combination with the NO donor molsidomine on plaque size, cell composition, superoxide production and extracellular superoxide dismutase (ecSOD) mRNA expression in the atherosclerotic plaques in the aorta. Cholesterol withdrawal alone did not alter atherosclerotic plaque size, the increased superoxide production or the decreased ecSOD mRNA, but led to the formation of a thin subendothelial macrophage-free layer and reduced both vascular cell adhesion molecule-1 expression and cell replication in the luminal part of the plaques. Treatment with molsidomine (1 mg/kg/day) during cholesterol withdrawal did not affect plaque size but increased the thickness of the subendothelial macrophage-free layer consisting of SMC, and normalized both superoxide production and ecSOD mRNA expression. The latter findings demonstrate that molsidomine, when combined with cholesterol lowering, decreases signs of oxidative stress and increases features of stable atherosclerotic plaques. PMID- 12775979 TI - Childhood bullous pemphigoid: a clinicopathologic study and review of the literature. AB - Bullous pemphigoid (BP) is an acquired bullous disorder that predominantly affects the elderly. It is rare in children but when it occurs, there is considerable clinical and histologic overlap with other acquired or congenital blistering disorders. A definitive diagnosis of childhood BP requires direct immunofluorescence and, in some cases, characterization of the target antigen. Three cases of childhood BP are presented, with their histologic and immunofluorescence findings. The first was a 5-month-old male infant who presented with erythema and bullae of the palms and soles and was found to have linear deposition of IgG and C3 along the dermoepidermal junction on direct immunofluorescence (DIF). Histopathologic examination revealed a subepidermal blister containing eosinophils. Type IV collagen was demonstrated along the floor of the blister cavity by a direct immunoperoxidase technique. The second case was an 8-month-old female infant who presented with a blistering eruption of her palms and soles that then became widespread. Direct immunofluorescence showed linear IgG and C3 at the dermoepidermal junction, with laminin deposition at the base of the blister. The third case was a 7-year-old female with bullae and erosions on the vulva and vaginal mucosa. A subepidermal blister was seen on microscopic examination whereas immunofluorescence demonstrated linear IgG and C3 deposition at the basement membrane zone (BMZ). A literature review uncovered 50 cases of childhood BP confirmed by direct or indirect immunofluorescence, or both, and often with evidence of autoantibodies against either the 180 kD or the 230 kD human bullous pemphigoid antigens (BP180 or BP230). This review was used to delineate characteristics of childhood BP, including the newly proposed subtypes: infantile BP and childhood localized vulval BP. Infantile BP presents within the first year of life and is characterized by BP-like lesions on erythematous or normal acral skin. Localized vulval BP is a self-limited, nonscarring BP-like process that involves only the vulva. Both subtypes are normally self-limited and respond well to either topical or systemic steroids, if treatment is initiated before the disease becomes widespread. PMID- 12775980 TI - Argyrophilic staining of nucleolar organizer region count and morphometry in benign and malignant melanocytic lesions. AB - Differentiation between malignant melanomas (MMs) and benign nevi based on histologic features can sometimes be difficult. This study evaluated the diagnostic effectiveness of argyrophilic staining of nucleolar organizer regions (AgNORs) in separating benign nevi from MMs by assessing 27 compound nevi (CN), 20 dysplastic nevi (DN), 10 Spitz nevi (SN), and 24 MMs. Both AgNOR count and morphology variables were measured from the superficial, middle, and deep zones of the lesions using video image analysis. Malignant melanomas had a significantly greater AgNOR number per nucleus, mean AgNOR area per nucleus, and variation in AgNOR area per nucleus compared with all types of benign nevi (p < 0.05). In multivariate discriminant analysis using a combination of four AgNOR counts and morphometric parameters, all CN and DN, 8 of 10 SN, and 23 of 24 MMs could be correctly classified as benign or malignant. The results suggest that both AgNOR count and morphology help to separate benign and malignant melanocytic lesions and that the combination of both sets of parameters improves their discriminating ability. PMID- 12775981 TI - A comparative study of immunohistochemistry and electron microscopy used in the diagnosis of epidermolysis bullosa. AB - Electron microscopic examination still is the gold standard for classifying epidermolysis bullosa, although it is relatively expensive, time consuming, and not readily available. Immunoreagents have been developed recently to map antigens in the basement membrane on routinely processed specimens. The current study was performed to examine the diagnostic usefulness of immunohistochemistry, as compared with electron microscopic examination, for analyzing routine formalin fixed paraffin-embedded sections of epidermolysis bullosa. This study investigated 39 consecutively diagnosed cases of epidermolysis bullosa in which both electron microscopic examination and immunohistochemistry were used. In each case, three monoclonal antibodies were used to stain for laminin 1, collagen IV, and keratin. The immunohistochemical patterns were defined as follows: epidermolysis bullosa simplex (laminin, collagen IV, or both at the dermal floor of the blister and keratin at both the dermal floor and the epidermal roof), junctional epidermolysis bullosa (laminin, collagen IV, or both at the dermal floor of the blister and keratin only at the epidermal roof), and dystrophic epidermolysis bullosa (collagen IV, laminin, or both, and keratin all at the epidermal roof). Altogether, electron microscopic examination subclassified epidermolysis bullosa into its three major forms in 37 of the 39 cases (95%), and immunohistochemistry in 33 of the 39 cases (85%). All of the classifiable cases were concordant. Specifically, immunohistochemistry was diagnostic in 10 of 14 (71%) epidermolysis bullosa simplex cases, 14 of 14 (100%) junctional epidermolysis bullosa cases, and 9 of 11 (82%) dystrophic epidermolysis bullosa cases. The most frequent cause for inconclusive immunohistochemical results was failure in staining of the basement membrane with the antibodies to both laminin and collagen IV. In conclusion, the use of immunohistochemistry on routinely processed specimens may be useful for subclassifying epidermolysis bullosa into its major forms in the majority of the cases, although it still cannot fully replace electron microscopic examination or immunofluorescence mapping in the diagnosis of epidermolysis bullosa. PMID- 12775982 TI - Nephrogenic fibrosing dermopathy after liver transplantation successfully treated with plasmapheresis. AB - Nephrogenic fibrosing dermopathy (NFD) is a recently described cutaneous fibrosing disorder associated with renal dysfunction. It appears similar to scleromyxedema but with some notable exceptions, including the lack of involvement of the face and absence of plasma cells on histology, systemic involvement, and paraproteinemia. Patients can present with thickened or edematous skin with indurated papules and plaques involving the extremities and the trunk. We report the first three cases of NFD after liver transplantation successfully treated with plasmapheresis. Two patients underwent liver transplantation for hepatitis C virus-induced cirrhosis and one for hepatitis B virus-induced cirrhosis. All the patients had encephalopathy, refractory ascites, and malnutrition prior to transplantation. Like those patients with NFD, all three of our patients had renal dysfunction and required hemodialysis before and after transplantation. Two were not dependent on dialysis at the time of diagnosis, however. These patients had excellent liver allograft function, but the other patient had allograft failure secondary to recurrent hepatitis C. Immunosuppression therapy consisted of basiliximab, mycophenolate mofetil, calcineurin inhibitor, and prednisone. The patients developed "woody" skin induration of the distal extremities, erythematous papules, and contractures at 1, 2, and 120 months after transplantation. Skin biopsies resembled NFD. No paraproteinemia was evident. One to three 5-day courses of plasmapheresis resulted in moderate to marked clinical improvement. The improvement of the kidney function in two of our patients did not appear to correlate with that of the skin disorder, because the kidney function was improving at the time the diagnosis of NFD was made. In conclusion, we report the first three cases of NFD after liver transplantation. Plasmapheresis was moderately successful in resolving the skin-indurated papules, severe skin induration, and associated joint contractures. Preliminary studies (unpublished data) show that decreasing plasma levels of transforming growth factor-beta1 after plasmapheresis appear to correlate with the amelioration of this clinical condition. PMID- 12775983 TI - Desmoplastic seborrheic keratosis. AB - Eleven cases of seborrheic keratoses with desmoplastic stroma and associated nests and cords of squamous epithelium simulating infiltrating carcinoma are presented. There were 7 males and 4 females ranging in age from 44 to 88 years (mean = 62). Eight cases were on the head and neck, one each on the lower and upper extremity, and one on the pubic area. Clinical diagnoses included seborrheic keratosis, squamous and basal cell carcinoma, and dermal nevus. Histologically, the lesions were characterized by exophytic growth pattern of basaloid and squamous cells, without cytologic atypia, and well-demarcated peripheral borders, typical for that seen in seborrheic keratoses. Squamous eddy formation with parakeratosis and spongiosis was present in all cases. Located within the body of the lesions were irregular nests and cords of squamous cells extending into the surrounding dermis with associated fibroblastic dermal proliferation trapping the epithelial nests, simulating invasive carcinoma. Immunohistochemically, the epithelial nests were cytokeratin positive and HPV negative, and the dermal stromal cells were vimentin positive and factor XIIIa, cytokeratin, and CD34 negative. Adjacent changes of carcinoma, trichilemmoma, verruca, or other adnexal neoplasm were not identified. Seborrheic keratoses may demonstrate desmoplastic changes analogous to that seen in desmoplastic trichilemmomas. Awareness of these changes in seborrheic keratosis will avoid misdiagnosis and excessive therapy. PMID- 12775984 TI - Tissue counter analysis of tissue components in skin biopsies: evaluation using CART (Classification and Regression Trees). AB - In tissue counter analysis, complex histologic sections are overlaid with regularly distributed measuring masks of equal size and shape, and the digital contents of each mask (or tissue element) are evaluated by gray level, color, and texture parameters. In this study, the feasibility of tissue counter analysis and classification and regression trees for the quantitative evaluation of skin biopsies was assessed. From 100 randomly selected skin biopsies, a learning set of tissue elements was created, differentiating between cellular elements, collagenous elements of the reticular dermis, fatty elements and other tissue components. Classification and regression trees based on the learning set were used to automatically classify tissue elements in samples of normal skin, benign common nevi, malignant melanoma, molluscum contagiosum, seborrheic keratosis, epidermoid cysts, basal cell carcinoma, and scleroderma. The procedure yielded reproducible assessments of the relative amounts of tissue components in various diagnostic groups. Furthermore, a reliable diagnostic separation of molluscum contagiosum versus normal skin and epidermal cysts, benign common nevi versus malignant melanoma, and seborrheic keratosis versus basal cell carcinoma was possible. Tissue counter analysis combined with classification and regression trees may be a suitable approach to the fully automated analysis of histologic sections of skin biopsies. PMID- 12775985 TI - Melanomas in prepubescent children: review comprehensively, critique historically, criteria diagnostically, and course biologically. AB - Our series was comprised of 11 children age 10 years or younger (6 were younger than age 5) with primary cutaneous melanoma. All of the melanomas occurred de novo and all metastasized; one child died. In no instance was melanoma a clinical consideration, and in none did the histopathologist who first "signed out" the case make a diagnosis of melanoma. Despite the inability of clinicians and pathologists to diagnose correctly, with repeatability, melanomas that develop in children yet to be pubescent, those neoplasms, nonetheless, are melanomas and, therefore, criteria employed currently for diagnosis of melanoma, especially clinically, must be refined in order that they be applicable equally to melanomas in pre- and postpubescents. The vaunted ABCDs (Asymmetry, Border irregular, Color variability, Diameter >6.0mm) surely do not work for melanomas that appear in children who are prepubescent. Additionally, melanomas that occur in these children have distinctly different architectural and cytopathological features from those that arise in postpubescents, often being confused as they are by conventional microscopy with a Spitz's nevus. As a rule, melanomas in prepubescent children grow much more rapidly then those in adults but, like them, have the capability to disseminate widely and cause death. PMID- 12775986 TI - Primary cutaneous ganglioneuroma: a report of two cases and literature review. AB - Primary cutaneous ganglioneuroma is a rare neoplasm first described by Collins et al. in 1972. Eight cases have been reported in the English literature to date. We report two cases of solitary cutaneous ganglioneuroma, one constituting, to our knowledge, the first reported case of this entity occurring on the face. Clinically, both lesions were firm flesh-colored papules and asymptomatic, being removed primarily for cosmesis. Neither patient had any significant past medical history. Histologically, both neoplasms consisted of relatively well circumscribed spindle cell proliferations of axons and Schwann cells with interspersed mature ganglion cells. There was also focal myxoid change. The spindle cell component stained positively for S-100 protein, and the ganglion cells stained positively for glial fibrillary acidic protein and neurofilaments by routine immunohistochemistry. PMID- 12775987 TI - Detection of human papillomavirus 60 in epidermal cysts of nonpalmoplantar location. AB - Human papillomavirus (HPV) types 57 and 60 are associated with epidermal cysts of palmoplantar location (PPECs). Recently, there was a report of HPV 60 in a wart located in a nonpalmoplantar area as a possible precursor of a nonpalmoplantar epidermal cyst (NPPEC). There has been no study that has examined the presence and frequency of HPV in ordinary NPPECs. We reviewed 63 cases of epidermal cysts, including 59 NPPECs and 4 PPECs. After routine histopathologic review, polymerase chain reaction (PCR) of extracted DNA, sequencing of PCR products, and, finally, a homology search were undertaken. As a result, 9 of 63 (14.3%) cysts, including 6 of 59 NPPECs (10.2%), showed a positive reaction on PCR. By homology search after PCR, all were demonstrated as HPV 60. For NPPECs, no variables, including some histopathologic features, are significantly related to the presence of HPV 60 in univariate or multivariate analysis (P > 0.05). PMID- 12775988 TI - Hyperplasia of the subcutaneous adipose tissue is the primary histopathologic abnormality in lipedematous scalp. AB - A 51-year-old white woman presented with thickening of the scalp located at the vertex and left lateral occiput without hair abnormalities or alopecia. Skin biopsies of the thickened scalp showed thickening of the subcutaneous tissue with proliferation of mature subcutaneous fat cells but no signs of inflammation or hair abnormalities. During 2.5 years of follow-up, scalp thickening progressed over the entire hair-bearing scalp and persisted without signs of further progression at 3.5 year follow-up. Lipedematous scalp is an extremely rare diagnosis. It is defined by a thickening of the subcutaneous layer of the scalp and can be distinguished from lipedematous alopecia, in which subcutaneous thickening is associated with diffuse alopecia and shortening of scalp hairs. A total of seven cases of lipedematous alopecia and two cases of lipedematous scalp have been reported. We report the third case of lipedematous scalp in a 51-year old white woman associated with early symptoms of meningitis. Additional features described in the literature include pruritus, pain, and paresthesia of the scalp as well as associated medical problems such as hyperelasticity of skin and laxity of joints, renal failure, and diabetes mellitus. This sporadic disorder is predominantly located at the vertex and occiput. The etiology and pathogenesis of lipedematous scalp and alopecia remain unclear. The treatment is symptomatic. PMID- 12775989 TI - Low-grade malignant eccrine spiradenoma with systemic metastases. AB - Malignant eccrine spiradenoma (MES) is a very rare tumor. Tumors can be low grade or high grade. There are nine cases in the literature with systemic metastases. Of these, only one was low grade; the others had obvious features of high-grade malignancy such as pleomorphism, increased mitoses, and abnormal mitoses. Low grade MESs are more difficult to diagnose and are said to have a better prognosis. We report an additional case of low-grade MES with systemic metastases. It had a monotonous appearance with few mitotic figures and minimal pleomorphism. Vascular invasion was seen in both cases of low-grade MES and may be an important prognostic feature. PMID- 12775990 TI - Was Henri Gougerot the first to describe "Hailey-Hailey Disease"? PMID- 12775991 TI - Trichoblastoma, basal cell carcinoma, and follicular differentiation: what should we trust? PMID- 12775992 TI - Criterion by criterion, mycosis fungoides. AB - A wide variety of histologic features have been associated with mycosis fungoides. Virtually all these features can be seen at least episodically in some inflammatory conditions, and little is known regarding which histologic features are most useful in establishing a diagnosis of this challenging condition. Three recent blind histologic studies have addressed this topic, however, and their results and those of other studies are reviewed and compared herein. PMID- 12775993 TI - Lupus erythematosus cells in the cutaneous lesion of overlap syndrome of dermatomyositis-SLE-like disease. PMID- 12775994 TI - Horn with miliary calcification in squamous cell carcinoma. PMID- 12775995 TI - Tailgut cyst as a subcutaneous tumor at the coccygeal region. PMID- 12775996 TI - Dermatitis herpetiformis with flame figures mimicking an arthropod bite. PMID- 12775997 TI - Impact of irritable bowel syndrome: prevalence and effect on health-related quality of life. AB - The prevalence of a disease and its effect on health-related quality of life (HRQOL) are important measures of its burden on society. The prevalence of irritable bowel syndrome (IBS) symptoms in the U.S. general population is approximately 10%, with lower estimates of prevalence if IBS is defined according to the more restrictive Rome II criteria. In population-based studies, there are no large differences in the prevalence of IBS symptoms between men and women or among the three major symptom subtypes of IBS (diarrhea- or constipation predominant or alternating). However, the majority of persons with IBS-like symptoms do not seek care for these symptoms and, in those who do seek care, there is a 2-to-1 female-to-male predominance. HRQOL is an important measure that should be considered in the overall assessment of a largely subjective, nonfatal disease such as IBS. Studies that have measured HRQOL in IBS used generic instruments, mostly the 36-Item Short Form Health Survey, and few have used IBS specific instruments. In a systematic review of the literature, there is strong evidence that persons with moderate to severe IBS who seek care for their symptoms (consulters) show decreased HRQOL. The impact of IBS on HRQOL in nonconsulters is less clear. Finally, a therapeutic response in IBS-related symptoms corresponds with an improvement in HRQOL. PMID- 12775998 TI - Diagnosing irritable bowel syndrome. AB - It is often possible to positively diagnose irritable bowel syndrome (IBS) based on a combination of multiple symptoms and their chronic nature. Both the Manning criteria and the ROME 1999 Consensus Working Party Diagnostic Criteria help in diagnosing IBS. It is important that, during the first visit, possible contributing factors, such as associated psychosocial stress or a history of mental, physical, or sexual abuse, are considered as part of the patient evaluation. Patients need to receive a clear explanation of the possible causes of symptoms, the benign nature of IBS, and the low likelihood of serious underlying disease. An interactive, positive physician-patient relationship has a beneficial effect on the course of IBS and may be associated with a decreased need for future health care visits. PMID- 12775999 TI - Traditional therapies for irritable bowel syndrome: an evidence-based appraisal. AB - Irritable bowel syndrome (IBS) is a common chronic disease that adversely effects quality of life and is associated with substantial direct and indirect health care costs. It is defined by a constellation of symptoms in conjunction with an alteration in bowel function and defecation, and its underlying pathophysiological basis remains unknown. Numerous therapies are available, but many relieve only one symptom of the syndrome, and their effectiveness has not been demonstrated with adequate evidence from high-quality studies. This article reviews the criteria for appropriate design of any treatment study as well as those criteria specific to studies of treatment for functional gastrointestinal diseases. Using these criteria, the author evaluates the published investigations of "traditional" IBS therapies (antidepressants, bulking agents/fiber, antispasmodics, antidiarrheals) and "alternative" IBS therapies (Chinese herbal supplements, peppermint oil, behavioral therapies). Based on this evaluation, the author concludes that the available evidence does not support the effectiveness of any of these treatments and, therefore, none of these treatments can be reliably recommended. PMID- 12776000 TI - Serotonin and its implication for the management of irritable bowel syndrome. AB - Our understanding of the enteric nervous system (ENS) has evolved from the "classical" view, in which the brain controls all enteric behavior, to the current view, which holds that enteric innervation is one of local control within the bowel, modified by a bidirectional "dialogue" with the brain. The ENS independently controls enteric reflexes through intrinsic primary afferent neurons, which monitor intraluminal conditions. This monitoring is accomplished through the use of enteroendocrine cells in the mucosa, the best known of which are the serotonin-containing enterochromaffin cells. This article describes the roles that serotonin, specific serotonin-receptor subtypes, and the serotonin reuptake transporter play in the ENS and in the communication between the ENS and central nervous system. The way in which these findings have implicated serotonin in irritable bowel syndrome is discussed. PMID- 12776001 TI - Tegaserod and other serotonergic agents: what is the evidence? AB - Through effects on gastrointestinal motor and secretory function as well as visceral sensation, serotonin (5-HT) plays a key role in the pathogenesis of irritable bowel syndrome (IBS). In particular, 5-HT3 and 5-HT4 receptors appear to be very important in IBS. This article critically appraises the evidence supporting the use of the 5-HT3 receptor antagonist alosetron in the treatment of women with diarrhea-predominant IBS. The safety profile and restricted-use program for alosetron is also reviewed. This discussion is followed by a comprehensive review of the efficacy and safety data in support of tegaserod for women with constipation-predominant IBS. PMID- 12776003 TI - Optimizing medical therapy for gastroesophageal reflux disease: state of the art. AB - Potential interventions for gastroesophageal reflux disease include lifestyle modifications, antacids, mucosal protectants, prokinetic (promotility) agents, H2 receptor antagonists (H2RAs) and, the agents of choice in 2003, proton pump inhibitors (PPIs). This article reviews the current state of the art in use of these agents. Lifestyle changes, though sound in their intent and in many cases based on solid laboratory research, can today be considered only adjuncts to pharmacologic therapy. The mainstay of pharmacologic therapy in 2003 is antisecretory therapy. Both H2RAs and PPIs inhibit acid secretion and raise intragastric pH. H2RAs only block one receptor, have limited effect on acid reduction, and are relatively weak inhibitors of meal-stimulated acid secretion. PPIs provide superior control of intragastric pH over a 24-hour period compared with H2RAs and effect greater symptom relief and healing. PMID- 12776004 TI - Registered nurse-administered propofol sedation for upper endoscopy and colonoscopy: Why? When? How? AB - Propofol for sedation during gastrointestinal endoscopic procedures has been associated with improved patient satisfaction relative to combinations of meperidine and midazolam. Delivery of propofol by registered nurses has been shown to be more cost-effective than administration by anesthesiologists or nurse anesthetists. Here, the authors review their experience with nurse-administered propofol sedation (NAPS) at their institutions (a hospital endoscopy unit in a tertiary medical center and an ambulatory surgery center). Endoscopic procedures for which NAPS is (or is not) recommended are listed, and a profile of patients for whom NAPS is contraindicated is given. Steps toward the development of programs and acquisition of training in NAPS are recommended; these include determining what state, local, and institutional laws apply to propofol administration; developing a written protocol; and instituting a training program that proceeds from observation to supervised administration to independent administration of propofol, with appropriate monitoring of safety records. Experience with NAPS using bolus titration (dosage, timing, coadministration with other agents) is detailed, and proper patient monitoring is discussed. NAPS is in its infancy, but this method of sedation shows considerable promise for improving patient satisfaction with, and thereby the reputation of, endoscopic procedures. PMID- 12776005 TI - The state of the art in the management of inflammatory bowel disease. AB - Ulcerative colitis (UC) and Crohn's disease (CD), collectively known as inflammatory bowel disease (IBD), afflict an estimated one million Americans and produce symptoms that impair quality of life and ability to function. Progress in IBD management strategies has led to optimized approaches for achieving the two primary clinical goals of therapy: induction and maintenance of remission. Although surgery is indicated to treat refractory disease or specific complications, pharmacotherapy is the cornerstone of IBD management. The efficacy of aminosalicylates for induction of remission in mild to moderate UC and CD is well established, as is their role for maintenance of remission in UC. The sulfa free mesalamine formulation offers an adverse effect profile similar to that of placebo, enabling the administration of higher, more effective doses. Although corticosteroids provide potent anti-inflammatory effects, their benefits are countermanded by the risk of intolerable and serious adverse effects, and they are ineffective for maintenance therapy. Other agents effective in inducing or maintaining remission are azathioprine, 6-mercaptopurine, infliximab, cyclosporine, methotrexate, and antibiotics. Ongoing clinical trials of experimental therapies will generate new tools for IBD treatment. Currently, a broad range of options allows physicians to tailor treatment to each patient's needs and preferences. Such considerations are essential for maximizing adherence to therapy. PMID- 12776006 TI - Pegylated interferon plus ribavirin for the treatment of chronic hepatitis C. AB - Peginterferon alfa-2a and peginterferon alfa-2b have been approved for the treatment of chronic hepatitis C virus (HCV) infection in adults who have compensated liver disease and have not been previously treated with interferon alfa. Peginterferon alfa-2a and peginterferon alfa-2b have also been approved for use in combination with ribavirin as therapy for these adults. Combining peginterferon alfa-2a or alfa-2b with ribavirin produces better activity against HCV than either drug alone. Interferon works by binding to specific receptors on the cell surface that initiate a complex cascade of protein-protein interactions leading to rapid activation of gene transcription. The effects of this interferon stimulated gene modulation depend on the biologic system and may result in the inhibition of viral replication in infected cells, inhibition of cell proliferation, and immunomodulation. Pegylation of the interferon molecule increases its size; the absorption of the larger pegylated molecule is slower, its half-life is longer, and its rate of clearance from the plasma is lower than that of the native interferon. Thus, the pegylated molecule increases the duration of biologic activity. Factors that appear to influence the success of pegylated interferon therapy are HCV genotype, baseline viral load, presence of fibrosis or inflammation shown on the liver biopsy at baseline, and the patient's body weight or body surface area. Patients infected with HCV genotype 1 tend to have a lower response rate, require longer courses of therapy, and respond better when treated with a pegylated interferon plus ribavirin. Patients infected with HCV genotypes 2 or 3 have comparable responses when treated with interferon plus ribavirin or pegylated interferon plus ribavirin and can be treated with a lower dose of ribavirin and a shorter course of therapy (24 weeks vs 48 weeks for patients with genotype 1). Studies directly comparing peginterferon alfa-2a and peginterferon alfa-2b have not been performed. PMID- 12776008 TI - Ventricular resynchronization: pathophysiology and identification of responders. AB - Patients with dilated cardiomyopathy and discoordinate wall motion due to intraventricular conduction delay are at increased risk for exacerbated pump failure and arrhythmias and suffer higher mortality rates. Biventricular and left ventricular resynchronization pacing therapies acutely improve systolic ventricular function and energetic efficiency in patients with heart failure and left-bundle-type intraventricular conduction delay. Sustained therapy can further inhibit or reverse chronic chamber dilation and remodeling. As with all therapies for heart failure, individual subject responses are variable; however, the invasive nature and expense of resynchronization therapy has particularly highlighted the need to prospectively identify optimal candidates. Although QRS duration has been principally used to date, increasing evidence shows this to have poor acute and chronic correlations with patient response. In contrast, direct measures of mechanical dyssynchrony based on simple echo imaging and more complex tissue Doppler and magnetic resonance imaging-based approaches appear to afford better predictive accuracy. PMID- 12776009 TI - Pathophysiology of congestive heart failure. AB - Heart failure is a clinical syndrome characterized by impaired structure and/or function of the heart, leading to dyspnea and fatigue at rest or with exertion. The pathophysiology of heart failure is complex, and there is no single lesion. Any form of heart disease can lead to heart failure. Most heart failure can be explained by well-recognized etiologic factors, though ostensibly healthy patients may harbor risk factors for the later development of heart failure. A fundamental response to myocardial injury or altered loading conditions includes "remodeling" of the heart, so that the size, shape, and function of the affected chamber is grossly distorted. This is accompanied by a constellation of biologic changes, best recognized in advanced cases of heart failure. These multiple alterations may be primary or secondary events but, nonetheless, add importantly to the morbidity and mortality of the patients. More emphasis should be placed on recognition and correction of risk factors related to the development of heart failure. PMID- 12776010 TI - Limitations of current medical therapies for the treatment of heart failure. AB - The medical treatment of heart failure has evolved over the past 40 years, from the primary use of diuretics and digitalis in the 1960s to the use of inotropic agents and vasodilators in the 1970s. More recently, the focus has been on the neurohormonal system, specifically the renin-angiotensin-aldosterone system and the sympathetic nervous system. Drugs that inhibit or block these systems (eg, angiotensin-converting enzyme inhibitors and beta-blocking drugs) are the primary agents recommended in recent heart failure guidelines. However, the actual percent reduction in mortality associated with the use of these agents has been relatively modest. This article will review the results and limitations of medical therapy for heart failure. Our understanding of the pathogenesis of heart failure may be incomplete, and alternative strategies, including mechanical devices, may play an increasing role in the treatment of heart failure in the future. PMID- 12776011 TI - Cardiac resynchronization therapy: a review of clinical trials and criteria for identifying the appropriate patient. AB - Numerous clinical trials have evaluated the safety and efficacy of cardiac resynchronization therapy in patients with moderate or severe heart failure and ventricular dyssynchrony. Initial studies were observational or mechanistic in design and enrolled small numbers of patients. These investigations provided proof of concept in support of resynchronization therapy by demonstrating acute and chronic improvements in hemodynamics, echocardiographic measures of cardiac performance, and functional status. Of these early studies, the InSync Trial stands out as particularly important in suggesting the long-term clinical benefits of cardiac resynchronization therapy in advanced heart failure. Later, a series of randomized, controlled trials was initiated to definitively evaluate the effects of cardiac resynchronization on patient status and clinical outcomes. These landmark investigations included the MUSTIC, MIRACLE, MIRACLE ICD, and CONTAK CD trials. These studies consistently demonstrated statistically significant improvements in quality of life, New York Heart Association (NYHA) functional class ranking, exercise tolerance, and left ventricular reverse remodeling. Some studies suggested reductions in morbidity and mortality. This latter observation was confirmed by a recent, large-scale morbidity and mortality trial of cardiac resynchronization therapy in heart failure. Given these findings, cardiac resynchronization therapy should be routinely considered in eligible NYHA Class III and IV heart failure patients with ventricular dyssynchrony. PMID- 12776012 TI - Cardiac resynchronization therapy devices: patient management and follow-up strategies. AB - Results of numerous clinical trials support the use of cardiac resynchronization therapy (CRT) for treatment of moderate to severe heart failure in patients with cardiac dyssynchrony. Commercial approval of various CRT devices has led to a growing number of patients receiving this form of therapy. Patients with implanted CRT devices require specific attention to detect adverse events related to the device, detect failure to respond to therapy, and optimize device function. Interaction between the primary care physician, cardiologist, heart failure specialist, and electrophysiologist/implanting physician is necessary to accomplish these goals. Recognizing the signs and symptoms of adverse events or suboptimal response allows the primary care physician to alter drug therapy or, when appropriate, refer the patient to the heart failure specialist or electrophysiologist for optimization of CRT device programming or further intervention. PMID- 12776013 TI - A guide to device selection: cardiac resynchronization therapy alone or in combination with an implantable cardioverter defibrillator. AB - The most recent estimates show an apparent increase in sudden cardiac death (SCD) in the United States. A major reduction in SCD will depend on developing effective methods to identify and prevent risk factors for SCD. This article reviews the research milestones that have resulted in our current approach to risk stratification and treatment of patients at high risk for SCD. One of the earliest attempts to prevent SCD involved suppression of premature ventricular complexes (PVCs) in high-risk patients, but trials using a variety of antiarrhythmic drugs with the aim of suppressing PVCs and reducing mortality demonstrated negative survival results. In the case of amiodarone, clinical trial data to date suggest that it should not be used for primary prevention of SCD or to prolong survival in patients with congestive heart failure secondary to coronary artery disease. The implantable cardioverter defibrillator (ICD) has been demonstrated in multiple studies to be the most significant therapy for life threatening ventricular arrhythmias and for primary and secondary prevention of SCD. It is recommended that the majority of patients who receive cardiac resynchronization therapy should have an ICD unit implanted in order to include defibrillator therapy. PMID- 12776014 TI - The role of stents in patients with carotid disease. AB - Carotid endarterectomy, the most commonly performed peripheral vascular surgical procedure, is associated with substantial clinical benefit when performed in patients with significant carotid bifurcation stenotic disease. The rate of morbidity and mortality, however, is significant when performed in patients with medical comorbidities or challenging surgical anatomy. Carotid stenting has emerged as a less invasive alternative to traditional endarterectomy. A large number of clinical trials, both randomized and registry-based, are ongoing. Initial results suggest that the outcome of carotid stenting may be identical to that of endarterectomy in most patients and may also offer additional clinical benefit in patients at high risk from an open surgical approach. PMID- 12776015 TI - The changing view of hormone replacement therapy. AB - Among postmenopausal women, 38% use hormone replacement therapy (HRT)- the leading brand of conjugated equine estrogen is the second most prescribed drug in the United States. Although adverse effects of this therapy have been recognized, it was traditionally reasoned that HRT could be broadly recommended to postmenopausal women because coronary heart disease was their most significant health risk. Evolving vascular biologic understanding, however, suggests that HRT, in direct opposition to the existing observational study data, does not reduce coronary heart disease risk. During the summer of 2002, three important studies on HRT dramatically changed our view of this widespread treatment. These trials provide clear, new, guidelines for patients and physicians contemplating HRT therapy. PMID- 12776016 TI - B-type natriuretic peptides: a diagnostic breakthrough for clinicians. AB - B-type natriuretic peptide (BNP), a neurohormone synthesized in the cardiac ventricles, is released as preproBNP and then enzymatically cleaved to the N terminal-proBNP (NT-proBNP) and BNP upon ventricular myocyte stretch. Blood measurements of BNP and NT-proBNP have been used to identify patients with congestive heart failure (CHF). Important considerations for these tests include their half-lives in plasma, dependence on renal function for clearance, and the interpretation of their units of measure. The BNP assay currently available in North American markets, approved for use as a diagnostic aid in CHF and a prognostic marker in acute coronary syndromes (ACS), has particular advantages because it is available at the point of care and has had considerable use in clinical studies. In general, a BNP level less than 100 pg/mL has strong negative predictive value for CHF. In addition, BNP levels can be used to gauge the effect of short-term treatment of acutely decompensated CHF. BNP has been shown to be a reliable and independent predictor of sudden cardiac death. In the absence of renal dysfunction, NT-proBNP has also been shown to be of diagnostic value in CHF, related to CHF severity, predictive of sudden death, and prognostic for death in ACS. This article reviews the literature concerning the use of these peptides in a variety of clinical scenarios. PMID- 12776017 TI - Diastolic function assessment incorporating new techniques in Doppler echocardiography. AB - Congestive heart failure (CHF) is the leading cause of cardiac morbidity and mortality from cardiovascular disease. Although left ventricular (LV) diastolic dysfunction occurs in all patients with systolic dysfunction and CHF, fully one third of patients have CHF due to isolated diastolic dysfunction. Despite this, the role of diastolic function in heart failure is underappreciated by many primary care physicians and even by cardiologists. Development and validation of several noninvasive Doppler echocardiographic techniques that are relatively load independent have made echocardiography the clinical standard for the assessment of LV diastolic function. Echocardiography uses portable equipment; is readily accessible and safe; and excludes other causes of CHF, such as valvular heart disease, pericardial disease, and systolic dysfunction. This review summarizes the use of recently developed Doppler techniques for the assessment of LV diastolic function, as well as their application in assessing prognosis and in guiding therapy for various cardiovascular disease states. PMID- 12776018 TI - Paradoxical coronary embolism: a rare cause of acute myocardial infarction. AB - Since it was first described in 1877, paradoxical embolism has become widely accepted as an etiology of stroke and peripheral vascular thrombotic occlusion. Less common are paradoxical emboli to the coronary artery. On the basis of limited pathologic and clinical series, it appears that paradoxical coronary emboli account for 5%-10% of all paradoxical emboli. Paradoxical coronary emboli have been described in all age groups, from neonates to the elderly, reflecting a wide spectrum of cardiac structural abnormality and clinical circumstance. Strategies for management should focus on treatment of acute coronary occlusion as well as prevention of future emboli. PMID- 12776019 TI - Transesophageal echocardiographic evaluation of perioperative systolic murmur in aortic pathology. AB - Development of a new systolic murmur in patients following a Bental procedure with a prosthetic or homograft aortic valve usually indicates an aortic valve related complication. Here, we report new etiologies of a loud systolic murmur in patients with aortic disease. One patient developed a new loud systolic murmur as an initial manifestation of acute type A aortic dissection without any complication, and two patients developed a loud systolic murmur as the major manifestation of aortic graft failure following aortic root surgery. Auscultation of a new loud systolic murmur in the upper chest in patients with known aortic disease should alert one to a complication within the ascending aorta. PMID- 12776025 TI - Tissue sampling and analysis. PMID- 12776026 TI - The role of endoscopy in the assessment and treatment of esophageal cancer. PMID- 12776027 TI - A case-control study of endoscopy and mortality from adenocarcinoma of the esophagus or gastric cardia in persons with GERD. AB - BACKGROUND: This study assessed whether EGD reduces mortality from adenocarcinoma of the esophagus or gastric cardia for patients with gastroesophageal reflux. METHODS: A case-control study was performed. A total of 245 incident cases of death caused by adenocarcinoma of the esophagus or gastric cardia (1995-1999) in which reflux was present were identified using Veterans Health Administration databases. A total of 980 controls with reflux but no death from adenocarcinoma were frequency matched for age, gender, and race. The occurrences of EGD from 1990 onward were compared for cases and controls. Logistic regression analysis with adjustment for potential confounding factors was performed. RESULTS: All the subjects were men. Cases were significantly less likely to have had an EGD in the time period of interest as compared with controls (adjusted odds ratio 0.66: 95% CI [0.45, 0.96], p = 0.03). This negative association was as strong for any EGD performed within 1 to 8 years before diagnosis as for a more recent EGD. However, there were no controls that included esophagectomy and no controls with a nonfatal diagnosis of adenocarcinoma, raising the question of whether EGD and reduced mortality are causally linked. The risk of dying from adenocarcinoma was significantly lower for men with a diagnosis of GERD as an inpatient relative to men in whom the diagnosis was made as an outpatient (adjusted odds ratio 0.21: 95% CI [0.15, 0.31], p < 0.01). CONCLUSIONS: For patients with GERD, performing an EGD is associated with reduced mortality from adenocarcinoma of the esophagus or gastric cardia, but whether this is a causative association remains unclear. PMID- 12776029 TI - PEG with introducer or pull method: a prospective randomized comparison. AB - BACKGROUND: PEG by the conventional pull method has the potential drawback of being associated with a higher frequency of wound infection, presumably caused by contamination of the gastrostomy catheter as it passes through the oral cavity. This study investigated the occurrence of peristomal wound infection after PEG placement by using the pull and introducer techniques. METHODS: Between September 1999 and May 2002, consecutive patients with dysphagia for whom PEG was recommended were enrolled in the study and randomly assigned to two groups: PEG with the introducer method (Group I) or PEG with the pull method (Group II). The peristomal area of each patient was evaluated on a daily basis for one week after PEG. Erythema and exudate were scored on a scale from 0 to 4 and induration on a scale of 0 to 3. Criteria for infection were a maximum combined score of 8 or higher, or the presence of microscopic and microbiologic evidence of suppurating exudate. In each group, the endoscope was passed once during the procedure, and an antibiotic (piperacillin) was given prophylactically. All procedures were performed by one investigator with the assistance of another physician. RESULTS: Of the 60 patients enrolled, 30 were assigned to each group. PEG was successful in all patients. One patient was excluded from each group because of death (Group I, stroke; Group II, myocardial infarction) within one week of the procedure. Therefore, 58 patients, 29 in each group, were evaluated. There was no significant difference between the groups in terms of clinical parameters (age, gender, disease, performance score, mode of previous feeding, and recent antibiotic exposure). The occurrence of peristomal infection within one week of PEG was lower in Group I (introducer method) (0 vs. 9; p = 0.00094). The mean daily combined scores in Group I were significantly lower than those in Group II. Median of maximum parameter scores in Group I were significantly lower than those in Group II. There were no procedure-related mortalities or clinically significant wound infections that required surgical intervention. CONCLUSIONS: The risk of peristomal wound infection after PEG is lower with the introducer method compared with the pull method. PMID- 12776028 TI - Multicenter randomized trial of the spiral Z-stent compared with the Wallstent for malignant biliary obstruction. AB - BACKGROUND: The industry standard since 1990 for self-expanding biliary metallic stents has been the Wallstent. In 1998 the Spiral Z-stent was released. This randomized trial compared the Z-stent with the Wallstent in the treatment of malignant biliary obstruction. METHODS: Patients with unresectable malignant biliary obstruction distal to the bile duct bifurcation were randomized to receive a 10-mm diameter Wallstent or a 10-mm diameter Z-stent. RESULTS: A total of 145 patients were randomized; 13 were excluded. Sixty-four patients who received a Z-stent and 68 who had a Wallstent are included in the analysis. Tumors responsible for bile duct obstruction were pancreatic cancer (108), cholangiocarcinoma (15), metastatic cancer (6), and papillary cancer (3). Metallic stents were successfully placed in all patients. Seven technical problems were encountered during placement of the Z-stent and 5 with the Wallstent. There were 21 occlusions requiring reintervention (8 Z-stent, 13 Wallstent; p = 0.30). Median time to reintervention was the following: Z-stent, 162 days; Wallstent, 150 days (p = 0.22). A total of 104 patients died of progressive disease or other cause; 7 patients remain alive with patent stents. The overall calculated median patency rates were: Z-stent, 152 days; Wallstent, 154 days (p = 0.90). CONCLUSIONS: The Spiral Z-stent is comparable with the Wallstent in terms of placement, occlusion rates, and overall patency. Occasional early occlusion of both stents suggests tumor characteristics instead of the size of the mesh openings in the stents as important factors. PMID- 12776030 TI - Conscious sedation for EUS of the esophagus and stomach: a double-blind, randomized, controlled trial comparing midazolam with placebo. AB - BACKGROUND: Patients undergoing EUS usually receive intravenously administered sedative medication. A double-blind, prospective, randomized trial was conducted to compare midazolam with a placebo for EUS of the esophagus and stomach and to assess patient tolerance and endoscopic feasibility. METHODS: A total of 111 patients were randomized to receive midazolam or a placebo. Patients used a visual analogue scale to score pre-EUS anxiety and tolerance for the procedure. Endoscopists used a visual analogue scale to score ease of echoendoscope introduction and overall patient cooperation. Subsequent to EUS, patients were asked if they were willing to undergo the procedure under the same conditions. RESULTS: The patients' visual analogue scale scores indicated that tolerance for the introduction of the echoendoscope and the procedure overall were both significantly better in the midazolam group. Overall patient tolerance was predicted by sedation (p < 0.001) and pre-EUS anxiety (p = 0.024). Endoscopists' visual analogue scale scores showed that ease of echoendoscope introduction was significantly better in the midazolam group. There was no significant difference in overall patient cooperation during the procedure. There were no significant differences in introduction time, total procedure time, or patients' willingness to undergo the procedure under the same conditions. CONCLUSIONS: EUS of the esophagus and stomach without intravenous sedation is feasible for endoscopists and patients. Patients prefer intravenous administration of midazolam. Assessment of anxiety before EUS may indicate whether administration of midazolam can improve tolerance for the individual patient. PMID- 12776031 TI - Effect of hysterectomy status on polyp detection rates at screening flexible sigmoidoscopy. AB - BACKGROUND: Flexible sigmoidoscopy with polypectomy reduces the incidence of colorectal cancer by removal of premalignant lesions. Factors that reduce the area examined by flexible sigmoidoscopy may reduce its benefit. The aim of this study was to determine whether hysterectomy affects completion and polyp detection rates at flexible sigmoidoscopy. METHODS: Within the setting of a multicenter, prospective, controlled trial of screening flexible sigmoidoscopy, patient and examination variables were compared by appropriate statistical methods for women between the ages of 55 and 64 years with and without a history of a hysterectomy. RESULTS: One quarter of women participants had undergone a hysterectomy. These women were more likely to have incomplete examinations (risk ratio [RR] of incomplete examination, 1.53; 95% CI [1.4, 1.6]). Flexible sigmoidoscopy was more difficult (p < 0.001), more painful (p < 0.001), and less extensive (46 cm vs. 48 cm insertion on average; p < 0.0001) in women who had undergone a hysterectomy. There was a significant trend toward lower relative detection rates of polyps and adenomas at more proximal sites (rectum, sigmoid colon, and proximal to sigmoid; respectively, p = 0.008, p = 0.009) in this group. CONCLUSIONS: Women who have undergone a hysterectomy have less extensive flexible sigmoidoscopy examinations, which are more difficult and more painful, than women without a hysterectomy. Hysterectomy is associated with a reduction in polyp detection rate in the sigmoid colon. This modality of screening may be less effective in women who have undergone a hysterectomy. PMID- 12776032 TI - Circumferential EMR and complete removal of Barrett's epithelium: a new approach to management of Barrett's esophagus containing high-grade intraepithelial neoplasia and intramucosal carcinoma. AB - BACKGROUND: There is no study of circumferential EMR in patients with Barrett's esophagus containing early stage malignant lesions. This study investigated the effectiveness and safety of circumferential EMR by using a simple snare technique without cap. METHOD: Patients with Barrett's esophagus containing multifocal high grade intraepithelial neoplasia or intramucosal cancer, and patients with endoscopically nonidentifiable early stage malignant mucosal changes incidentally detected in random biopsy specimens were included in the study. A 30 x 50-mm polypectomy snare made of monofilament 0.4-mm steel wire was used without any additional device or submucosal injection. RESULTS: Twelve patients (10 men, 2 women; median age 63.5 years, range 43-88 years) underwent circumferential EMR; 5 had multifocal lesions, and 7 had no visible lesions. Segments of Barrett's epithelium were circumferential (median length 5 cm) and completely removed. The median number of EMR sessions was 2.5. The median number of snare resections per EMR session was 5. The medial total area of mucosa in resected specimens per session was 3.8 cm(2). Two patients developed strictures that were successfully treated by bougienage. Minor bleeding occurred during 4 of 31 EMR sessions. During a median follow-up of 9 months, no recurrence of Barrett's esophagus or malignancy was observed. CONCLUSIONS: Circumferential EMR with a simple snare technique is feasible, safe, and effective for complete removal of Barrett's epithelium with early stage malignant changes. PMID- 12776033 TI - Phase II study of photodynamic therapy and metal stent as palliative treatment for nonresectable hilar cholangiocarcinoma. AB - BACKGROUND: The combination of photodynamic therapy and biliary drainage by plastic endoprosthesis insertion has produced promising results in the treatment of nonresectable hilar cholangiocarcinoma. The feasibility and efficacy of intraductal photodynamic therapy with subsequent biliary drainage by self expandable metal stent insertion were evaluated in a prospective phase II study. METHODS: Twenty-four patients were treated with photodynamic therapy after sensitization with porfimer sodium. A plastic endoprosthesis was inserted immediately thereafter and replaced by a metal stent 4 weeks later. A retrospectively analyzed group of 20 patients treated only with biliary drainage served as a historical control group. RESULTS: In 19 of the 24 patients, insertion of a metal stent was technically feasible. The 30-day and 60-day mortality rates were 0%. A significant decrease in serum bilirubin was noted in all patients and quality of life remained stable throughout follow-up. Mean and median survival were, respectively, 15.9(3.1) and 9.9: 95% CI [6.4, 13.4] months after photodynamic therapy. In the control group, mean and median survival were, respectively, 12.5(3.4) and 5.6: 95% CI [3.7, 7.6] months, which was not statistically significantly different from the photodynamic therapy group. CONCLUSIONS: Photodynamic therapy with consecutive biliary drainage by insertion of a self-expandable metal stent is feasible. With respect to the small benefit in overall survival, randomized controlled trials are warranted. PMID- 12776034 TI - Understanding endoluminal gastroplications: a histopathologic analysis of intraluminal suture plications. AB - BACKGROUND: Endoluminal gastroplication is used to treat GERD, with modest results. Little is known of the histologic reaction to endosutures. Thus, the histologic response to intraluminal plications at different penetration depths within the gastric wall was studied in an animal model. METHODS: Intraluminal gastroplications were performed through a laparotomy in 18 New Zealand rabbits. Three sets of everted plications were placed at different penetration depths in each stomach: submucosa (SMpl), muscularis propria (Mpl), and serosa (Spl). Animals were randomized to survival times of 3, 10, or 60 days (respectively, Groups I, II, and III). Plications were compared with a grade scale for each histologic healing phase and gross inspection. RESULTS: Fusion between folds was absent in all groups. Serosa differed from muscularis propria with respect to the proportion of samples with microscopic ischemia (67% vs. 8%; p = 0.015), remaining sutures in Group III (33% vs. 3%; p < 0.05 in a single test of significance, but correction for multiple testing removes this significance), and remaining plications in Group II (96% vs. 54%; p < 0.05 in a single test of significance, but correction for multiple testing removes this significance). All of the total and partial histologic scores for the corresponding healing phase in each group escalated with penetration depth. Overall comparison of the histologic scores showed a significant difference among the plications in the proliferation (Group II, p = 0.004) and maturation (Group II, p = 0.009) phases. Total scores also differed among the plications in Groups II (p < 0.001) and III (p < 0.001). Plications were absent in all of Group III, with Spl resulting only in a flat scar. CONCLUSION: Everted intraluminal gastroplications do not result in fusion between folds irrespective of suture-penetration depth. A flat scar is the final outcome and appears proportional to the amount of ischemia, foreign body reaction, and suture depth. PMID- 12776035 TI - Diagnostic yield of routine push enteroscopy with a graded-stiffness enteroscope without overtube. AB - BACKGROUND: Push enteroscopy has become a standard procedure for evaluation of small intestinal disorders. Its diagnostic yield and acceptability, however, has been hampered by the use of an overtube, which is both inconvenient and potentially hazardous. This study assessed the clinical value of enteroscopy with a graded-stiffness videoenteroscope without an overtube. METHODS: A total of 121 consecutive patients (mean age 59 years, range 12-89 years) underwent diagnostic enteroscopy. All procedures (n = 126) were performed with a push-type graded stiffness videoenteroscope without an overtube. Indications were the following: unexplained iron deficiency anemia (45%), GI bleeding (29%), abdominal pain (6%), malabsorption (5.5%), imaging abnormality (5.5%), diarrhea (4%), intestinal obstruction (3%), and vomiting (2%). RESULTS: The mean depth of instrument insertion distal to the pylorus was 121 cm. A diagnosis was made in 40% of all procedures. The findings included ulcerations or erosions in 43%, angioectasia in 35%, inflammation in 14%, tumors in 6%, and varices in 2%. In all cases of a positive enteroscopic diagnosis, therapeutic maneuvers were performed, and no patient needed a further diagnostic procedure. Patient comfort was good. No complications were observed. CONCLUSIONS: Routine enteroscopy with a graded stiffness enteroscope without an overtube is safe and comfortable for the patient and the endoscopist, and has a clinical efficacy comparable with that reported for enteroscopy with use of an overtube. A prospective, randomized study is warranted to assess the exact role of this form of enteroscopy in patient care. PMID- 12776036 TI - Morbidity and mortality after self-expandable metallic stent placement in patients with progressive or recurrent esophageal cancer after chemoradiotherapy. AB - BACKGROUND: Placemet of self-expandable metallic stents in patients with advance esophageal cancer improves dysphagia and occludes tracheoesophageal fistulas. However, the safety of self-expandable metallic stents for patients who have undergone chemoradiotherapy is controversial. This study evaluated the morbidity and modality after self-expandable metallic stent placement in patients with progressive or recurrent esophageal cancer after chemoradiotherapy. METHODS: A total of 22 patients in whom self-expandable metallic stents were placed because of progressive or recurrent esophageal cancer after chemoradiotherapy were studied. RESULTS: All 22 patients had dysphagia, and 13 had a tracheoesophageal fistula. After self-expandable metallic stent placement, the mean dysphagia grade improved from 3.5 to 0.9, and tracheoesophageal fistula was successfully managed in all cases. Seventeen patients had T4 stage disease, and among 8 of them with invasion to the aorta, 6 (75%) died of sudden massive hemorrhage. Median survival for these 6 patients was 31 days (range 13-63 days) compared with 67 days (range 4-262 days) for all patients after self-expandable metallic stent placement. CONCLUSION: Self-expandable metallic stent placement improved dysphagia and was useful for treatment of tracheoesophageal fistula. However, for patients with T4 lesions that invade to the aorta, self-expandable metallic stent placement after chemoradiotherapy should be considered carefully. PMID- 12776037 TI - A direct comparison of ERCP teaching models. AB - BACKGROUND: Several teaching models for ERCP are now available. Live, anesthetized porcine models have been used for many years, but harvested porcine organ preparations have recently been developed, and computer-based endoscopy simulators now incorporate ERCP modules. Each has proven to be a useful educational modality, but there is no direct comparison among these models. This study compared the performance of these 3 ERCP teaching models. METHODS: Twenty endoscopists used each ERCP training model (computer simulator, harvested porcine organ, live anesthetized pig) and then completed a survey grading the realism and performance of each model compared with performance of ERCP in patients. A rank order was established for the models relative to their realism, educational utility, ease of use, and ease of incorporation into a training program. RESULTS: The harvested porcine organ model scored highest on indices of realism, usefulness, and performance, although this reached statistical significance only for "ease of use" (p < 0.05). Conversely, the computer simulator scored significantly lower in most realism scores, although it was felt to be the one model most easily incorporated into a training program. CONCLUSIONS: Although each ERCP teaching model has proven to be a useful training modality, the harvested porcine organ model was felt to be the most realistic as well as the most favorable model for instruction in both basic and advanced ERCP. PMID- 12776038 TI - EUS in the evaluation of pancreatic cystic lesions. AB - BACKGROUND: The differential diagnosis in pancreatic cystic lesions is often difficult despite the availability of various modern imaging modalities. This study assessed the role of EUS in the following: (1) discrimination of pseudocysts from pancreatic cystic tumors, (2) differential diagnosis between serous cystadenoma and mucinous cystic tumor, and (3) prediction of accompanying malignancy in intraductal papillary mucinous tumor. METHODS: EUS findings in 75 patients with pancreatic cystic lesions (58 cystic tumors, 17 pseudocysts) were evaluated. In the comparison of pseudocysts and cystic tumors, the latter included intraductal papillary mucinous tumor, mucinous cystic tumors, and serous cystadenomas, but not solid-pseudopapillary tumors. RESULTS: In univariate analysis, pseudocysts exhibited echogenic debris and parenchymal changes more often than cystic tumors did (respectively, 29% vs. 6%, p < 0.05; and 65% vs. 4%, p < 0.001). In contrast, septa and mural nodules were found more frequently in cystic tumors than pseudocysts (respectively, 69% vs. 12%, p < 0.001; 56% vs. 12%, p < 0.01). Multivariate analysis revealed that parenchymal changes (odds ratio [OR] = 83.59; p < 0.01); septa (OR = 30.75; p < 0.05); and mural nodules (OR = 21.38; p < 0.05) were independent predictors of differentiation between pseudocysts and cystic tumors. Serous cystadenoma exhibited diverse EUS features, as well as a honeycomb appearance. Mural nodules were found more often in mucinous cystic tumors than in serous cystadenomas (p < 0.05). There were no factors that predicted malignancy in intraductal papillary mucinous tumor. CONCLUSIONS: EUS is a useful complementary imaging method for differentiation of pancreatic cystic lesions. PMID- 12776039 TI - Mucosal ablation with photodynamic therapy in the esophagus: optimization of light dosimetry in the sheep model. AB - BACKGROUND: Photodynamic therapy is an attractive technique for mucosal ablation in patients with superficial squamous cell carcinoma of the esophagus, or high grade dysplasia or early stage adenocarcinoma arising in Barrett's esophagus. Although illumination with green light is assumed to be safe, choice of the light has been empirical in clinical studies; light dose is often reduced to avoid potential complications. The present study assessed the safety of green and blue lights during photodynamic therapy in the esophagus by progressively administrating increasing doses in an attempt to standardize the dose and determine a safe upper limit. This would considerably simplify photodynamic therapy and improve therapeutic results. METHODS: The sheep model was chosen because of similarities with humans regarding the thickness and histologic structure of the esophagus. Irradiation with a 180 degrees windowed cylindrical light distributor was performed in 19 sheep 4 days after injection of 0.15 mg/kg of tetra(m-hydroxyphenyl) chlorin. Light doses ranged from 10 to 500 J/cm(2) at 514 nm (argon ion laser) and from 5 to 250 J/cm(2) at 413 nm (krypton laser). RESULTS: Follow-up endoscopies revealed a tissue response with a fibrinous area at almost all light doses, whereas application of extremely high light doses tended to induce circumferential necrosis with subsequent stenosis. Three months after irradiation with green light, histologic examination of the resected specimens revealed transmural scarring at doses higher than 100 J/cm(2). After illumination with blue light, partial or more extensive fibrosis of the muscular layer was observed only at light doses of 175 to 250 J/cm(2). CONCLUSIONS: Application of high doses of green light after sensitization with tetra(m hydroxyphenyl) chlorin led to severe complications in the esophagus of the sheep that are highly likely to occur in humans as well. Blue light causes significantly less damage than green light and may, therefore, be considered as an alternative for photodynamic therapy of early stage superficial esophageal cancer. PMID- 12776040 TI - Giardiasis. PMID- 12776041 TI - Endoscopic screening and surveillance for Barrett's esophagus: can claims data determine its effectiveness? PMID- 12776042 TI - Cronkhite Canada syndrome. PMID- 12776043 TI - Biliary diverticula in sclerosing cholangitis. PMID- 12776044 TI - Cystic Brunner's gland hamartoma. PMID- 12776045 TI - Gastric and duodenal ischemia. PMID- 12776046 TI - Complete esophagogastric anastomotic disruption. PMID- 12776047 TI - Bleeding rectal Dieulafoy's lesion. PMID- 12776048 TI - EUS-guided celiac plexus neurolysis and celiac plexus block. PMID- 12776049 TI - A comparison of image quality between tissue harmonic imaging and fundamental imaging with an electronic radial scanning echoendoscope in the diagnosis of pancreatic diseases. AB - BACKGROUND: The availability of an electronic radial scanning echoendoscope has facilitated the clinical use of tissue harmonic imaging. This study compares the quality of US images acquired by tissue harmonic imaging during electronic radial scanning EUS to those acquired by fundamental imaging. METHODS: Electronic radial scanning EUS was performed in 108 patients with pancreatic lesions (58 cystic, 50 solid). US images acquired by fundamental imaging at a frequency of 7.5 MHz were compared with those acquired by tissue harmonic imaging by using transmitting and receiving frequencies of, respectively, 4.0 and 8.0 MHz at the same scanning plane. Cystic lesions were evaluated for boundary/septum and nodules, and solid lesions, for boundary and internal structure. US images acquired by tissue harmonic imaging and fundamental imaging during electronic radial scanning EUS were compared, by using a Likert scale for the respective evaluation criteria, by two independent endoscopists. OBSERVATIONS: For cystic lesions, tissue harmonic images were significantly clearer than fundamental images for visualizing boundary and septum (p < 0.0001, both reviewers) and nodules (p = 0.0003, Reviewer 1; p = 0.0007, Reviewer 2). For solid lesions, tissue harmonic images were significantly clearer than fundamental images for visualizing boundary (p = 0.0003, Reviewer 1; p < 0.0001, Reviewer 2) and internal structures (p = 0.0003, Reviewer 1; p = 0.0009, Reviewer 2). CONCLUSIONS: US images acquired by tissue harmonic imaging appear to be clearer compared with those acquired by fundamental imaging. PMID- 12776050 TI - Evaluation of a new curved linear array echoendoscopy system for EUS. AB - BACKGROUND: EUS-FNA can be used to accurately diagnose and stage GI and pulmonary neoplasms. This study evaluated the performance characteristics of a new compact linear EUS system during EUS-FNA. METHODS: A total of 37 patients enrolled in this prospective pilot study underwent clinically indicated EUS-FNA and/or celiac plexus neurolysis with the compact EUS system. RESULTS: The mean time to perform a radial and linear array EUS with FNA and/or celiac plexus neurolysis was 24 minutes shorter with the compact EUS system compared with that for an historical control procedure in which a conventional linear EUS unit was used (p = 0.0007). The EUS images and visualization of the needle during EUS-FNA were rated good to excellent in greater than 95% of the patients. With respect to ease of esophageal intubation, duodenal intubation, and general maneuverability, the performance of the new linear echoendoscope, compared with a radial scanning videoechoendoscope, was the same or better in, respectively, 85%, 87%, and 100% of procedures. The video image quality of the new linear array echoendoscope was superior to that of the radial scanning videoechoendoscope in all patients. No complications were encountered. CONCLUSIONS: EUS-FNA and celiac plexus neurolysis can be performed safely with the new compact EUS system. The efficiency of this procedure is enhanced compared with historical experience with other instruments. Although image quality is sufficient for EUS-FNA and celiac plexus neurolysis, the compact unit cannot be used as a "stand-alone" system for routine diagnostic EUS, and its use must be complemented by standard radial imaging. PMID- 12776051 TI - Use of a 19-gauge injection needle as a guide for direct percutaneous endoscopic jejunostomy tube placement. AB - BACKGROUND: Direct percutaneous endoscopic jejunostomy (DPEJ) tube placement is becoming an accepted means of achieving enteral nutrition. However, during DPEJ placement it can be difficult to maintain the position of the small bowel for insertion of the plastic sheath with stylet, thus limiting the success of the procedure. The results of a technique designed to overcome this problem are presented. METHODS: During DPEJ placement, a 19-gauge injection needle was passed into the bowel at the site of transillumination. The needle was snared tightly, fixing the small bowel against the abdominal wall. The plastic sheath with stylet was then inserted adjacent to the 19-gauge needle and into the small bowel and was subsequently snared to facilitate guidewire passage. A 24F, pull-type PEJ tube was then placed in standard fashion. RESULTS: A DPEJ was placed successfully in 24 of 26 (92.3%) patients with this technique. There was one (4%) major complication: inadvertent small bowel perforation during DPEJ placement. The average time to complete a procedure was 23.3 minutes; the mean time to achieve the dietary goal after DPEJ placement was 39 hours. One patient died of an unrelated illness 6 days after DPEJ placement; 23 were discharged with jejunal feeding. CONCLUSIONS: A DPEJ can be performed successfully by using a 19-gauge injection needle as guide for tube placement. PMID- 12776052 TI - A new technique for removal of bile duct stones with an expandable metallic stent. AB - BACKGROUND: Endoscopic sphincterotomy is used routinely for extraction of bile duct stones. Also, endoscopic papillary dilation is a safe and effective technique that significantly reduces the need for papillotomy. However, extraction of large and/or multiple stones after endoscopic papillary dilation can be difficult. A new technique, endoscopic metallic stent-lithotripsy, for treatment of bile duct stones without endoscopic sphincterotomy or endoscopic papillary dilation is described. METHODS: A self-expandable metallic stent was used to dilate the major duodenal papilla to allow lithotripsy and removal of bile duct stones in 38 patients. RESULTS: The bile duct was successfully cleared of stones in 36 cases (95%). Complications included one episode of mild pancreatitis and one of cholangitis. CONCLUSIONS: Although the number of patients who underwent successful expanding metallic stent-lithotripsy was small, the method is promising as an alternative to endoscopic sphincterotomy and endoscopic papillary dilation. PMID- 12776053 TI - Endoscopic closure of perforations caused by EMR in the stomach by application of metallic clips. AB - BACKGROUND: The number of complications associated with use of EMR for early stage gastric cancer, including perforation, has increased with the increasing use of this procedure. Endoscopic clip application was performed in patients who sustained a perforation as a result of EMR for gastric neoplasm. PATIENTS AND METHODS: Seven patients who underwent endoscopic application of metallic clips to close perforations were studied. The omental patch method was applied in one case with a large perforation. OBSERVATIONS: In all patients, endoscopic clip application successfully closed the perforation of the stomach, which occurred after EMR. No patient required laparotomy. CONCLUSIONS: The technique of endoscopic clip application might be useful for treatment of patients who sustain a perforation caused by EMR. PMID- 12776054 TI - Epstein-Barr virus-negative gastric large B-cell lymphoma after kidney transplantation. PMID- 12776055 TI - EMR of mucosa-associated lymphoid tissue lymphoma of the rectum. PMID- 12776056 TI - Gastric wall abscess presenting as a submucosal tumor: case report. PMID- 12776057 TI - Endoscopic clip application for closure of an esophagomediastinal-tracheal fistula after surgery for esophageal cancer. PMID- 12776058 TI - Diagnosis of solid-pseudopapillary neoplasm of the pancreas by EUS-guided FNA. PMID- 12776059 TI - Gastric cancer associated with Dieulafoy's lesion: case report. PMID- 12776060 TI - Successful treatment of an undifferentiated early stage gastric cancer by combined en bloc EMR and laparoscopic regional lymphadenectomy. PMID- 12776061 TI - Acute bleeding and anemia associated with intraluminal duodenal diverticulum: case report and review. PMID- 12776062 TI - Early stage esophageal carcinoma in an epiphrenic diverticulum. PMID- 12776063 TI - Anterograde colonoscopy: per oral diagnosis of colon cancer with an enteroscope in a man with a jejunoileal bypass. PMID- 12776064 TI - Esophageal submucosal gland duct adenoma: characteristic EUS and histopathologic features. PMID- 12776065 TI - Spontaneous bile leak 6 years after uneventful cholecystectomy. PMID- 12776066 TI - Pseudomalignant erosion in an inflammatory polyp at esophagocardial junction. PMID- 12776067 TI - Endoscopic removal of a spoon from the stomach with a double-snare and balloon. PMID- 12776068 TI - Synchronous and metachronous lesions may occur in patients with celiac disease anywhere in the GI tract. PMID- 12776069 TI - Tips to make band ligation affordable. PMID- 12776070 TI - Nursing's perfect storm--staff shortages and patient ratios. PMID- 12776071 TI - Current affairs: heroes, SARS, staffing, and family presence. PMID- 12776072 TI - Are there any laws or regulations to prevent critically ill inpatients from being brought down to an already overcrowded emergency department for care? PMID- 12776074 TI - Form to track patients with frequent ED visits. PMID- 12776075 TI - A 12-month-old girl with maculopapular lesions and lower extremity edema. PMID- 12776076 TI - Family presence during cardiopulmonary resuscitation and invasive procedures: practices of critical care and emergency nurses. AB - BACKGROUND: Increasingly, patients' families are remaining with them during cardiopulmonary resuscitation and invasive procedures, but this practice remains controversial and little is known about the practices of critical care and emergency nurses related to family presence. OBJECTIVE: To identify the policies, preferences, and practices of critical care and emergency nurses for having patients' families present during resuscitation and invasive procedures. METHODS: A 30-item survey was mailed to a random sample of 1500 members of the American Association Of Critical-Care Nurses and 1500 members of the Emergency Nurses Association. RESULTS: Among the 984 respondents, 5% worked on units with written policies allowing family presence during both resuscitation and invasive procedures and 45% and 51%, respectively, worked on units that allowed it without written policies during resuscitation or during invasive procedures. Some respondents preferred written policies allowing family presence (37% for resuscitation, 35% for invasive procedures), whereas others preferred unwritten policies allowing it (39% for resuscitation, 41% for invasive procedures). Many respondents had taken family members to the bedside (36% for resuscitation, 44% for invasive procedures) or would do so in the future (21% for resuscitation, 18% for invasive procedures), and family members often asked to be present (31% for resuscitation, 61% for invasive procedures). CONCLUSIONS: Nearly all respondents have no written policies for family presence yet most have done (or would do) it, prefer it be allowed, and are confronted with requests from family members to be present. Written policies or guidelines for family presence during resuscitation and invasive procedures are recommended. PMID- 12776077 TI - Responding to the severe acute respiratory syndrome (SARS) outbreak: lessons learned in a Toronto emergency department. PMID- 12776078 TI - Increased volume/length of stay for pediatric mental health patients: one ED's response. PMID- 12776079 TI - Methotrexate administration for ectopic pregnancy in the emergency department- one hospital's protocol/competencies. PMID- 12776080 TI - ENA's new guidelines for determining emergency department nurse staffing. PMID- 12776081 TI - Knowledge assessment and preparation for the Certified Emergency Nurses Examination. PMID- 12776082 TI - Orienting ED nurses to triage: using scenario-based test-style questions to promote critical thinking. PMID- 12776083 TI - An informal discussion of emergency nurses' current clinical practice: what's new and what works. PMID- 12776084 TI - Top issues in EMS as discussed by leading industry experts. PMID- 12776085 TI - Recreational sports: a Northeast Wisconsin hospital's approach in reducing morbidity and mortality. PMID- 12776087 TI - Some practical tips for more effective teaching. PMID- 12776088 TI - Second impact syndrome: a rare, catastrophic, preventable complication of concussion in young athletes. PMID- 12776089 TI - Small amounts of some drugs can be toxic to young children: one pill or one swallow can require aggressive treatment. PMID- 12776090 TI - Clostridium difficile colitis following an open fracture: complications occur, even with straightforward trauma and straightforward decisions. PMID- 12776091 TI - A 2-year-old patient with fever and vomiting. PMID- 12776092 TI - Microfabricated adhesive mimicking gecko foot-hair. AB - The amazing climbing ability of geckos has attracted the interest of philosophers and scientists alike for centuries. However, only in the past few years has progress been made in understanding the mechanism behind this ability, which relies on submicrometre keratin hairs covering the soles of geckos. Each hair produces a miniscule force approximately 10(-7) N (due to van der Waals and/or capillary interactions) but millions of hairs acting together create a formidable adhesion of approximately 10 N x cm(-2): sufficient to keep geckos firmly on their feet, even when upside down on a glass ceiling. It is very tempting to create a new type of adhesive by mimicking the gecko mechanism. Here we report on a prototype of such 'gecko tape' made by microfabrication of dense arrays of flexible plastic pillars, the geometry of which is optimized to ensure their collective adhesion. Our approach shows a way to manufacture self-cleaning, re attachable dry adhesives, although problems related to their durability and mass production are yet to be resolved. PMID- 12776093 TI - It's not rocket science--or is it? PMID- 12776094 TI - Surfaces--lost and found. PMID- 12776096 TI - Biomaterials: thwarting thrombus. PMID- 12776097 TI - Optoelectronics: a deep-level LED. PMID- 12776098 TI - Organic electronics: molecules as bipolar conductors. PMID- 12776099 TI - Solar cells: a solid compromise. PMID- 12776100 TI - Ionic liquids: a taste of the future. PMID- 12776101 TI - Material witness: patent pending. PMID- 12776102 TI - Chiral selection on inorganic crystalline surfaces. AB - From synthetic drugs to biodegradable plastics to the origin of life, the chiral selection of molecules presents both daunting challenges and significant opportunities in materials science. Among the most promising, yet little explored, avenues for chiral molecular discrimination is adsorption on chiral crystalline surfaces - periodic environments that can select, concentrate and possibly even organize molecules into polymers and other macromolecular structures. Here we review experimental and theoretical approaches to chiral selection on inorganic crystalline surfaces - research that is poised to open this new frontier in understanding and exploiting surface-molecule interactions. PMID- 12776103 TI - HUGO--a UN for the human genome. PMID- 12776105 TI - Adherence to published ethical guidelines by the UK genetics research community. PMID- 12776106 TI - MEDLINE definitions of race and ethnicity and their application to genetic research. PMID- 12776110 TI - Turned off by RNA. PMID- 12776111 TI - Epigenetic interplay. PMID- 12776112 TI - The advantages of recombination. PMID- 12776115 TI - When calcium goes wrong: genetic alterations of a ubiquitous signaling route. AB - In all eukaryotic cells, the cytosolic concentration of calcium ions ([Ca2+]c) is tightly controlled by complex interactions among transporters, pumps, channels and binding proteins. Finely tuned changes in [Ca2+]c modulate a variety of intracellular functions, and disruption of Ca2+ handling leads to cell death. Here we review the human genetic diseases associated with perturbations in the Ca2+ signaling machinery. Despite the importance of Ca2+ in physiology and pathology, the number of known genetic diseases that can be attributed to defects in proteins directly involved in Ca2+ homeostasis is limited to few examples, which will be discussed. This paucity in contrast with the wide molecular repertoire may depend on the extreme severity of the phenotype (leading to death in utero) or, conversely, on functional compensation due to redundancy. In the latter case, it stands to reason that other genetic defects in calcium signaling have yet to be identified owing to their subtle phenotype. PMID- 12776118 TI - Whither RNAi? PMID- 12776120 TI - The return of the exocyst. PMID- 12776121 TI - Reproductive cloning conserves cellular senescence. PMID- 12776122 TI - Nuclear pore assembly: locating the linchpin. PMID- 12776123 TI - The cutting edge of mitochondrial fusion. PMID- 12776124 TI - Deconstructing phosphatases in limb development. PMID- 12776125 TI - Meddling with methylation. PMID- 12776128 TI - Hybrid rigor mortis. PMID- 12776127 TI - Polar expeditions--provisioning the centrosome for mitosis. AB - It is now clear that both centrioles and their surrounding pericentriolar material (PCM) are capable of self-assembly. Whereas centrioles are normally duplicated during G1-S phase, PCM components may be loaded onto centrosomes in both a microtubule-dependent and -independent manner at all stages of the cell cycle. Centrosomes enlarge dramatically after mitotic entry, when both Aurora A and Polo-like kinases cooperate to recruit additional gamma-tubulin ring complexes and microtubule-associated proteins to assist spindle formation. PMID- 12776130 TI - Germany contemplates new biotech tax scheme. PMID- 12776131 TI - EU stalls on funding of ES cell research. PMID- 12776132 TI - Agbiotech climbs Africa's agenda. PMID- 12776133 TI - US FDA contemplates collection of pharmacogenomic data. PMID- 12776134 TI - India dawdles over Bt-cotton. PMID- 12776136 TI - Clarifying press before paper. PMID- 12776137 TI - The challenge of industrializing proteomics. PMID- 12776138 TI - Identity tags. PMID- 12776139 TI - Making 'safety first' a reality for biotechnology products. PMID- 12776140 TI - Putting nature back into drug discovery. PMID- 12776141 TI - Hidden in plain view. PMID- 12776143 TI - Public biotechnology 2002--the numbers. PMID- 12776144 TI - US venture capital for biotechnology. PMID- 12776145 TI - Value creation and sharing among universities, biotechnology and pharma. PMID- 12776146 TI - Targeting mammalian genes--rats join in and mice move ahead. PMID- 12776147 TI - Mass spectrometry provides sweet inspiration. PMID- 12776148 TI - RNAi puts a lid on virus replication. PMID- 12776149 TI - Plugging metal connectors into enzymes. PMID- 12776151 TI - Protecting rights to early-stage technology. PMID- 12776155 TI - New immigration laws affect US biotechnology. PMID- 12776156 TI - Gene therapy progress and prospects: gene therapy for the hemophilias. AB - Recent gene transfer trials for hemophilia A and B, bleeding disorders lacking either functional factor VIII or IX, respectively, have produced tantalizing results, suggesting that the potential to correct these bleeding disorders at a molecular level may be at hand. Genetic correction of the hemophilias represents a model system to develop a basic understanding of how gene therapy will be achieved. The goals for hemophilia gene transfer require the long-term therapeutic production of the coagulant protein without stimulating an immune response to the transgene product or the vector. Based on a scientific understanding of the molecular and cellular defects, leading to the bleeding phenotype, impressive strides have been made in the last 2 years. PMID- 12776158 TI - Titer determination of Ad5 in blood: a cautionary note. AB - Recombinant adenoviruses are presently the most efficient in vivo gene transfer system available. Targeting single organs or large tumors by adenoviral vectors requires an intravascular route of application. During the first pass of viral particles through the vascular bed of the target tissue, virus uptake is not quantitative and indefinite amounts of particles leak into circulation. To determine the amount of leaking particles and to calculate organ-specific uptake (in-/outflow ratio), it is necessary to titrate virus particles directly in blood. In preclinical and clinical trials titration is currently mostly done with blood plasma instead of full blood. However, this technique provides valid results only as long as there is no affinity between adenovirus particles and erythrocytes. In this study we demonstrate that Ad5 particles, as mostly employed for gene therapy, have a strong affinity to human erythrocytes. At 60 min after coincubation of human erythrocytes and Ad5 particles, more than 98% of the particles are attached to the surface of erythrocytes. Therefore, ignoring the amount of red cell bound particles by performing titration in plasma leads to severe miscalculation of organ-specific transfer rates or virus circulation half life. The biological impact of an increased affinity between virus particles and erythrocytes will be discussed. PMID- 12776157 TI - Adenoviral delivery of IL-18 binding protein C ameliorates collagen-induced arthritis in mice. AB - Elevated concentrations of interleukin-18 (IL-18) are found in both serum and synovial fluid of patients suffering from rheumatoid arthritis (RA) and this cytokine has recently been implicated in the development of experimental arthritis. In this present study, we developed an IL-18 neutralizing intervention and examined its efficacy for local intra-articular treatment of experimental arthritis. To this end we constructed an adenoviral vector containing the murine IL-18 binding protein isoform c gene (AdCMVIL-18BPc). The constructed adenoviral vector was validated on replication deficiency, transfection efficacy and ability to express biological functional IL-18BPc. Intra-articular overexpression of IL 18BPc significantly reduced incidence of collagen-induced arthritis (CIA) in treated kneejoints. Affected kneejoints of IL-18BPc-treated mice showed less severe arthritis, characterized by reduction of inflammation and destruction of bone and cartilage. Local intra-articular IL-1BPc treatment in both knees provided additional protection against CIA incidence and severity in distal paws. Measurement of serum levels of specific collagen type (CII) Abs revealed a moderate reduction of circulating IgG2a anti-CII Abs, while IgG1 anti-CII Abs remained at similar level. The present study underlines the involvement of IL-18 as an important proinflammatory cytokine in onset of experimental arthritis. Furthermore, it shows that endogenous IL-18 can be blocked efficiently through local adenoviral overexpression of IL-18BPc, indicating that treatment with IL 18BPc might contribute to joint protection in RA. PMID- 12776159 TI - Reversion of transformed phenotype in ovarian cancer cells by intracellular expression of anti folate receptor antibodies. AB - The alpha-folate receptor (FR) is selectively overexpressed in 90% of nonmucinous ovarian carcinomas, whereas no expression is detectable in normal ovarian surface epithelium (OSE). Indirect evidence suggests that FR expression is associated with tumor progression and affects cell proliferation. To evaluate better the role of FR, we developed an approach based on intracellular expression of single chain (sc) antibodies (intrabody) to downmodulate membrane expression of FR in ovary cancer cells. IGROV-1 and SKOV3 ovarian carcinoma cell lines were transfected with an anti-FR intrabody. Transfectants and parental cells were tested for FR, integrins and anti-FR intrabody expression by fluorescence activated cell sorting (FACS), reverse transcription and polymerase chain reaction (RT-PCR) and/or immunoblotting. Cell growth characteristics and adhesion properties were evaluated in liquid, semisolid and organotypic cultures. The anti FR scFv inhibited FR expression from 60 to 99%. At physiological concentrations of folate, proliferation varied directly as a function of FR expression. FR downmodulation was accompanied by reduced colony-forming ability in soft agar, morphological change of the cells, significant enhanced adhesion to laminin or Matrigel, a two- to three-fold increase in alpha6beta4 integrin expression, and a marked reduction in laminin production. In three-dimensional organotypic cultures, anti-FR intrabody-transfected IGROV1 cells grew as a single-ordered layer, reminiscent of normal OSE growth in vivo. In conclusion, the anti-FR intrabody reverses the transformed phenotype in ovary cancer cells and may provide an efficient means to inhibit selectively the growth of these cells. PMID- 12776160 TI - Formation of LID vector complexes in water alters physicochemical properties and enhances pulmonary gene expression in vivo. AB - There is currently an urgent need to develop efficient gene-delivery systems for the lung that are free of inflammatory effects. The LID vector is a synthetic gene delivery system, comprised of lipofectin (L), an integrin-targeting peptide (I) and DNA (D) that has previously been shown to have high transfection efficiency in the lung. We have assessed the effect of alternative methods of complex preparation on structural features of the complex, levels and duration of reporter gene expression and the host response to the LID vector. We have demonstrated that making the complex in water affects the structure of the LID complexes making them smaller and more stable with a more cationic surface charge than complexes prepared in phosphate-buffered saline (PBS). When the LID vector was constituted in water and instilled intratracheally into the lungs of mice there was a 10-fold increase in luciferase activity compared with preparation in PBS. Furthermore, luciferase activity was still evident 1 week following vector instillation. This enhancement may be because of altered complex structure, although effects of the hypotonic vector solution on the lung cannot be excluded. The inflammatory effects of instilling the LID vector in water were minimal, even after three administrations of the LID vector, with only mild alterations in cytokine and broncho-alveolar lavage fluid (BALF) cell profiles. These results demonstrate that the LID vector can generate high, and prolonged, levels of gene expression in the lung from small quantities of DNA and that careful attention to synthetic polyplex structure may be important to optimize efficiency of gene expression in vivo. PMID- 12776161 TI - Transcriptional targeting of dendritic cells for gene therapy using the promoter of the cytoskeletal protein fascin. AB - Strong cell-type-specific promoters are basic tools in gene therapy allowing for novel applications and focused strategies by transcriptionally targeting gene expression to selected cells. In immunotherapy, dendritic cells (DC) are of central importance, since they represent the principal inducers of immune responses. Here we describe isolation and use of the promoter of the murine actin bundling protein fascin to target transcriptionally gene expression to cutaneous DC. Using the reporter gene enhanced green fluorescent protein (EGFP), we demonstrate that the fascin promoter mediates a strong antigen expression that is restricted to mature DC. DNA vaccination with antigen-encoding expression vectors under control of the fascin promoter using a gene gun resulted, consistently, in limited antigen expression by few directly transfected DC. Nevertheless, nearly as many antigen-specific CD8+ T cells directed against the encoded antigens EGFP and beta-galactosidase, respectively, were induced as with expression constructs under control of the ubiquitously expressed CMV promoter. This result impressively underlines the pivotal role of directly transfected DC in DNA vaccination. Immunization using the fascin promoter induced markedly lower levels of antigen-specific antibodies following single or repeated immunization. Thus, our DC-targeted DNA vaccination approach induces qualitatively distinct, predominantly cellular immune responses and provides new opportunities for immunotherapy. PMID- 12776162 TI - Efficient gene transfer into human CD34+ cells by an adenovirus type 35 vector. AB - Efficient gene transfer into human hematopoietic stem cells (HSCs) is the most important requirement for gene therapy of hematopoietic disorders and for study of the hematopoietic system. An adenovirus (Ad) vector based on the Ad serotype 5 (Ad5) is known to transduce HSCs, including CD34(+) cells, with very low efficiency because of low-level expression of its primary receptor, coxsackievirus and adenovirus receptor (CAR). In the present study, we developed a recombinant Ad vector composed of the whole Ad serotype 35 (Ad35), which recognizes an unidentified receptor different from CAR for its infection. A transduction study showed that the Ad35-based vectors exhibit a higher transduction efficiency in human CD34(+) cells than the conventional Ad5 vectors and the Ad5F35 vectors, which are fiber-substituted Ad5 vectors containing Ad35 fiber proteins. The mean of fluorescence intensity in the CD34(+) cells transduced with the Ad35 vectors was 12-76 and 1.4-3 times higher than that in the cells transduced with the Ad5 and Ad5F35 vectors, respectively. The percentages of green fluorescent protein (GFP)-positive CD34(+) cells by transduction with Ad35, Ad5, and Ad5F35 vectors expressing GFP at 300 PFU/cell were 53%, 5%, and 52%, respectively, suggesting that Ad35 vectors mediate a more efficient gene transfer into human CD34(+) cells than Ad5 and Ad5F35 vectors, although the percentage of transduced cells was similar between Ad35 and Ad5F35 vectors. The Ad vector based on Ad35 could be very useful in gene therapy for blood disorders and gene transfer experiments using HSCs. PMID- 12776163 TI - Hypoxia-regulated transgene expression in experimental retinal and choroidal neovascularization. AB - Recombinant AAV vectors mediate efficient and sustained transgene expression in retinal tissues and offer a powerful approach to the local, sustained delivery of angiostatic proteins for the treatment of ocular neovascular disorders. The application of such strategies may also require regulated gene expression to minimize the potential for unwanted adverse effects. In this study, we have evaluated the effect of a hypoxia-responsive element (HRE) on the kinetics of recombinant adeno-associated (rAAV)-mediated reporter gene expression in murine models of retinal and choroidal neovascularization. In murine ischaemia-induced retinal neovascularization, intravitreal delivery of rAAV.HRE.GFP results in reporter gene expression specifically at sites of vascular closure during the period of active neovascularization and not after vector delivery in normal controls. In murine laser-induced choroidal neovascularization, subretinal delivery of rAAV.HRE.GFP results in reporter gene expression at sites of active neovascularization but not elsewhere or after vector delivery in normal controls. HRE-driven gene expression offers an attractive strategy for the targeted and regulated delivery of angiostatic proteins to the retina in the management of neovascular disorders. PMID- 12776164 TI - Intra-articular adenoviral-mediated gene transfer of trail induces apoptosis of arthritic rabbit synovium. AB - Rheumatoid arthritis (RA) is an inflammatory autoimmune disease that primarily affects joints. In rheumatoid joints there is extensive synovial proliferation with diseased synovium becoming highly aggressive, attaching to the articular cartilage and bone to form what is termed a pannus. The formation of active pannus is central to erosive disease and resulting joint destruction. In this study, we examined the ability to eliminate the hyperplastic synovium by adenoviral-mediated gene transfer of human TNF-related apoptosis-inducing ligand (TRAIL), a member of the TNF family that is able to induce apoptosis through interaction with receptors containing death domains, DR4 and DR5. Infection of synovial cells derived from RA patients with Ad.TRAIL resulted in significant apoptosis in three out of five lines. Moreover, primary rabbit synovial fibroblasts were also sensitive to Ad.TRAIL-mediated gene transfer. In a rabbit model of arthritis, intra-articular gene transfer of TRAIL induced apoptosis in cells within the synovial lining, reduced leukocytic infiltration and stimulated new matrix synthesis by cartilage. These results demonstrate that TRAIL can affect the viability of the cells populating the activated synovium in arthritic joints and suggest that the delivery of TRAIL to arthritic joints may represent a non-invasive mechanism for inducing pannus regression. PMID- 12776166 TI - Patriotism and publication. PMID- 12776165 TI - Impact of splice-site mutations of the human MDR1 cDNA on its stability and expression following retroviral gene transfer. AB - The multidrug resistance 1 (MDR1) gene transfer to hematopoietic cells for protection against cytotoxic drugs has received considerable attention in gene therapy. However, ectopic expression of MDR1 from retroviral vectors has been hampered by its genetic instability resulting from cryptic splice sites within the cDNA. We have evaluated the efficiency of retroviral MDR1 vectors with introduced mutations of the MDR1 cryptic splice donor (cSD) located at nucleotide +339 and of the cryptic splice acceptor (cSA) at nucleotide +2319 of the cDNA. Sequence alterations of the cSD reduced the expression of MDR1 P-glycoprotein (P gp), even when generated as silent mutations. A silent mutation of the cSA reduced the splicing activity shifting the splice acceptor site one base downstream; however, it significantly improved the expression of P-gp. The incidence of wild-type MDR1 pregenome splicing was markedly reduced when vectors were produced in human 293 packaging cells as opposed to murine PG13 and GP+envAm12. We conclude that complete splice correction of MDR1 in retroviral vectors may only be achieved with extensive alterations of the cDNA or neighboring vector sequences and that the splicing is significantly influenced by the choice of the packaging cells. PMID- 12776167 TI - Comment on Levanon et al., "Runx3 knockouts and stomach cancer", in EMBO reports (June 2003). PMID- 12776168 TI - The transformation of the biological sciences in post-war Europe. PMID- 12776169 TI - Footing the bill. PMID- 12776170 TI - Aliens on the shores. PMID- 12776171 TI - Gulf War syndrome revisited. PMID- 12776172 TI - Local solutions for global problems. PMID- 12776173 TI - From cells on up: Symposium on Cell Biology of Development and Disease. PMID- 12776174 TI - Runx3 knockouts and stomach cancer. AB - Gene targeting often results in knockout mice that show several phenotypes, some of which may not directly relate to the intrinsic function of the disrupted gene. Hence, to study the biological function of genes using knockout mice, one must identify the defects that are directly due to the loss of the targeted gene. Runx3 is a transcription factor that regulates lineage-specific gene expression in developmental processes. Recently, two groups produced Runx3 knockout mice. Two comparable defects were identified in both knockout strains, one involved neurogenesis and the other thymopoiesis. In addition, a stomach defect pertaining to gastric cancer was observed in one of the mutant strains, but not in the other. Here, we assess the differences between the two Runx3 mutant strains and discuss further studies that could reconcile these discrepancies. This article highlights the difficulties of inferring gene function through the interpretation of knockout phenotypes. PMID- 12776175 TI - Getting a grip on non-native proteins. AB - It is an underappreciated fact that non-native polypeptides are prevalent in the cellular environment. Native proteins have the folded structure, assembled state and cellular localization required for activity. By contrast, non-native proteins lack function and are particularly prone to aggregation because hydrophobic residues that are normally buried are exposed on their surfaces. These unstable entities include polypeptides that are undergoing synthesis, transport to and translocation across membranes, and those that are unfolded before degradation. Non-native proteins are normal, biologically relevant components of a healthy cell, except in cases in which their misfolding results from disease-causing mutations or adverse extrinsic factors. Here, we explore the nature and occurrence of non-native proteins, and describe the diverse families of molecular chaperones and coordinated cellular responses that have evolved to prevent their misfolding and aggregation, thereby maintaining quality control over these potentially damaging protein species. PMID- 12776176 TI - IQGAP proteins are integral components of cytoskeletal regulation. AB - IQGAP1 is a scaffolding protein that binds to a diverse array of signalling and structural molecules. By interacting with its target proteins, human IQGAP1 participates in multiple cellular functions, including Ca(2+)/calmodulin signalling, cytoskeletal architecture, CDC42 and Rac signalling, E-cadherin mediated cell-cell adhesion and beta-catenin-mediated transcription. Yeast IQGAP homologues are important regulators of cellular morphogenesis because they are required for budding and cytokinesis. Here we discuss the structure and function of IQGAP1 as a member of the family of IQGAP proteins and summarize the current knowledge about IQGAP1 and IQGAP2. Collectively, these data reveal that IQGAP1 is a fundamental regulator of cytoskeletal function. PMID- 12776177 TI - MYC recruits the TIP60 histone acetyltransferase complex to chromatin. AB - The transcription factor MYC binds specific DNA sites in cellular chromatin and induces the acetylation of histones H3 and H4. However, the histone acetyltransferases (HATs) that are responsible for these modifications have not yet been identified. MYC associates with TRRAP, a subunit of distinct macromolecular complexes that contain the HATs GCN5/PCAF or TIP60. Although the association of MYC with GCN5 has been shown, its interaction with TIP60 has never been analysed. Here, we show that MYC associates with TIP60 and recruits it to chromatin in vivo with four other components of the TIP60 complex: TRRAP, p400, TIP48 and TIP49. Overexpression of enzymatically inactive TIP60 delays the MYC induced acetylation of histone H4, and also reduces the level of MYC binding to chromatin. Thus, the TIP60 HAT complex is recruited to MYC-target genes and, probably with other other HATs, contributes to histone acetylation in response to mitogenic signals. PMID- 12776178 TI - Effects of ligand and thyroid hormone receptor isoforms on hepatic gene expression profiles of thyroid hormone receptor knockout mice. AB - Little is known about the overall patterns of thyroid hormone (Th)-mediated gene regulation by the main Th receptor (Tr) isoforms, Tr-alpha and Tr-beta, in vivo. We used 48 complementary DNA microarrays to examine hepatic gene expression profiles of wild-type and Thra and Thrb knockout mice under different Th conditions: no treatment, treatment with 3,3',5-triiodothyronine (T(3)), Th deprivation using propylthiouracil (PTU), and treatment with a combination of PTU and T(3). Hierarchical clustering analyses showed that positively regulated genes fit into three main expression patterns. In addition, only a subpopulation of target genes repressed basal transcription in the absence of ligand. Interestingly, Thra and Thrb knockout mice showed similar gene expression patterns to wild-type mice, suggesting that these isoforms co-regulate most hepatic target genes. Differences in the gene expression patterns of Thra/Thrb double-knockout mice and Th-deprived wild-type mice show that absence of receptor and of hormone can have different effects. This large-scale study of hormonal regulation reveals the functions of Th and of Tr isoforms in the regulation of gene expression patterns. PMID- 12776179 TI - Targeting mortalin using conventional and RNA-helicase-coupled hammerhead ribozymes. AB - Mortalin, also known as mot2/mthsp70/GRP75/PBP74, is a member of the heat-shock protein 70 family that is heat-uninducible. It is differentially distributed in cells that have normal and immortal phenotypes, has been localized to various subcellular sites, and has several binding partners and functions. Here, we describe the construction and use of mortalin-specific conventional and hybrid ribozymes to elucidate its crucial role in cell proliferation. Whereas conventional hammerhead ribozymes did not cause any repression of endogenous mortalin expression, RNA-helicase-linked hybrid ribozymes successfully suppressed the expression of mortalin, which resulted in the growth arrest of transformed human cells. We show that, first, RNA helicase-coupled hybrid ribozymes that have a linked unwinding activity can be used to target genes for which conventional hammerhead ribozymes are ineffective; second, the targeting of mortalin by RNA helicase-coupled hybrid ribozymes causes growth suppression of transformed human cells and could be used as a treatment for cancer. PMID- 12776180 TI - Specific inhibition of gene expression using a stably integrated, inducible small interfering-RNA vector. AB - We have designed a doxycycline-regulated form of the H1 promoter of RNA polymerase III that allows the inducible knockdown of gene expression by small interfering RNAs (siRNAs). As a proof-of-principle, we have targeted beta-catenin in colorectal cancer (CRC) cells. T-cell factor (TCF) target-gene expression is induced by accumulated beta-catenin, and is the main transforming event in these cells. We have shown previously that the disruption of beta-catenin/TCF4 activity in CRC cells by the overexpression of dominant-negative TCF induces rapid G1 arrest and differentiation. Stable integration of our inducible siRNA vector allowed the rapid production of siRNAs on doxycycline induction, followed by specific downregulation of beta-catenin. In these CRC cells, TCF reporter-gene activity was inhibited, and G1 arrest and differentiation occurred. The inhibition of two other genes using this vector system shows that it should be useful for the inducible knockdown of gene expression. PMID- 12776181 TI - Interaction between the small-nuclear-RNA cap hypermethylase and the spinal muscular atrophy protein, survival of motor neuron. AB - The biogenesis of spliceosomal small nuclear ribonucleoproteins (snRNPs) requires the cytoplasmic assembly of the Sm-core complex, followed by the hypermethylation of the small nuclear RNA (snRNA) 5' cap. Both the Sm-core complex and the snRNA trimethylguanosine cap are required for the efficient nuclear import of snRNPs. Here, we show that trimethylguanosine synthase 1 (TGS1), the human homologue of the yeast snRNA cap hypermethylase, interacts directly with the survival of motor neuron (SMN) protein. Both proteins are similarly distributed, localizing in the cytoplasm and in nuclear Cajal bodies. The interaction between TGS1 and SMN is disrupted by a mutation in SMN that mimics the predominant isoform of the protein that is expressed in patients with the neurodegenerative disease, spinal muscular atrophy. These data indicate that, in addition to its function in cytoplasmic Sm core assembly, the SMN protein also functions in the recruitment of the snRNA cap hypermethylase. PMID- 12776182 TI - The role of the death-domain kinase RIP in tumour-necrosis-factor-induced activation of mitogen-activated protein kinases. AB - The death-domain kinase RIP (receptor-interacting protein) is an important effector of tumour necrosis factor (TNF) signalling and is essential for TNF induced nuclear factor-kappaB activation. However, the function of RIP in the TNF induced activation of mitogen-activated protein kinases (MAPKs) has not been fully investigated. In this report, using Rip null (Rip(-/-)) mouse fibroblast cells, we investigated whether RIP is required for TNF-induced activation of the MAPKs extracellular-signal-related kinase (ERK), p38 and c-Jun amino-terminal kinase (JNK). We found that TNF-induced activation of ERK, p38 and JNK is decreased in Rip(-/-) cells. The activation of these kinases by interleukin-1 is normal in Rip(-/-) cells. More importantly, we showed that the kinase activity of RIP is needed for ERK activation. PMID- 12776183 TI - O-mannosylation precedes and potentially controls the N-glycosylation of a yeast cell wall glycoprotein. AB - Secretory proteins in yeast are N- and O-glycosylated while they enter the endoplasmic reticulum. N-glycosylation is initiated by the oligosaccharyl transferase complex and O-mannosylation is initiated by distinct O mannosyltransferase complexes of the protein mannosyl transferase Pmt1/Pmt2 and Pmt4 families. Using covalently linked cell-wall protein 5 (Ccw5) as a model, we show that the Pmt4 and Pmt1/Pmt2 mannosyltransferases glycosylate different domains of the Ccw5 protein, thereby mannosylating several consecutive serine and threonine residues. In addition, it is shown that O-mannosylation by Pmt4 prevents N-glycosylation by blocking the hydroxy amino acid of the single N glycosylation site present in Ccw5. These data prove that the O- and N glycosylation machineries compete for Ccw5; therefore O-mannosylation by Pmt4 precedes N-glycosylation. PMID- 12776184 TI - Human arteries engineered in vitro. AB - There is a pressing need to develop methods to engineer small-calibre arteries for bypass surgery. We hypothesized that the rate-limiting step that has thwarted previous attempts to engineer such vessels from non-neonatal tissues is the limited proliferative capacity of smooth muscle cells (SMCs), which are the main cellular component of these vessels. Ectopic expression of the human telomerase reverse transcriptase subunit (hTERT) has been shown recently to extend the lifespan of certain human cells. We therefore introduced hTERT into human SMCs and found that the resulting cells proliferated far beyond their normal lifespan but retained characteristics of normal control SMCs. Importantly, using these non neonatal SMCs, we were able to engineer mechanically robust human vessels, a crucial step towards creating arteries of clinical value for bypass surgery. PMID- 12776186 TI - Retinoic acid-induced growth arrest of MCF-7 cells involves the selective regulation of the IRS-1/PI 3-kinase/AKT pathway. AB - In the MCF-7 breast cancer cell line, insulin-like growth factors (IGFs) are known to elicit antiproliferative actions via the insulin receptor substrate-1 (IRS-1)/PI 3-kinase/AKT pathway. All-trans retinoic acid (RA) is a potent inhibitor of MCF-7 cell proliferation, but the mechanism by which growth regulation is achieved remains unclear. We investigated the effects of RA on the regulation of the IGF-IR and its key signaling elements: IRS-1, IRS-2, and SHC. Treatment of MCF-7 cells with RA caused a significant reduction in IRS-1 protein and tyrosine phosphorylation levels at a concentration and time consistent with RA-mediated growth inhibition. IRS-1 regulation is selective, as RA did not influence IRS-2 or SHC levels. Downstream signaling events were also selectively reduced, as RA abrogated IGF-I-stimulated AKT activation but did not alter erk1/2 activation. To confirm the importance of IRS-1 regulation by RA, we examined the response to RA in MCF-7 cells overexpressing IGF-IR and IRS-1. RA resistance was observed in MCF-7 cells overexpressing IRS-1 but not IGF-IR. This suggests that RA-mediated growth inhibition requires the selective downregulation of IRS-1 and AKT. Therapeutic agents targeting the IRS-1/PI 3-kinase/AKT pathway may enhance the cytostatic effects of RA in breast cancer, since overexpression of IRS-1 and AKT have been reported in primary breast tumors. PMID- 12776187 TI - VEGF receptor expression and signaling in human bladder tumors. AB - Overexpression of vascular endothelial growth factor receptors (VEGFRs) has been reported in a variety of tumor types. Here we find that 11 out of the 14 bladder tumor cell lines examined express one or more VEGF receptors. Analysis of the T24 bladder tumor cell line reveals a functional autocrine loop involving VEGF and the Flk-1 receptor. Blocking VEGF expression in T24 cells results in a decrease in DNA synthesis. The Flk-1 receptor in T24 cells is phosphorylated in response to VEGF-121 or VEGF-165, and an Flk-1 inhibitor blocks VEGF to ERK signaling. We report that VEGF stimulation of T24 cells results in activation of H- and N-Ras and this is dependent on cellular sphingosine kinase 1 (SPK1) activity. Previously, we found VEGF-induced activation of Ras appears to be independent of a Ras-guanine nucleotide exchange factors (GEFs). Here we report that sphingosine can stimulate Ras-GTPase activating protein (GAP) activity in vitro, and sphingosine-1-phosphate (SPP) can block the stimulatory effects of sphingosine. We present a model where the balance between sphingosine and SPP regulates Ras GAP activity such that stimulation of SPK1 favors downregulation of Ras-GAP and thereby the activation of Ras proteins. These data highlight a VEGF pathway that may be involved in the survival and proliferation of bladder tumor cells as well as other tumor cell types. PMID- 12776188 TI - Kaposi's sarcoma-associated herpesvirus encoded vFLIP induces cellular IL-6 expression: the role of the NF-kappaB and JNK/AP1 pathways. AB - The Kaposi's sarcoma-associated herpesvirus (KSHV) encodes a FADD-like interferon converting enzyme or caspase 8 (FLICE) inhibitory protein (vFLIP) that prevents death receptor-mediated apoptosis by inhibiting the recruitment and activation of FLICE. Since vFLIP physically interacts with tumor necrosis factor receptor associated factor 2 (TRAF2) and TRAF2 mediates activation of the jun NH(2) terminal kinase (JNK)/activation protein 1 (AP1) pathway, we hypothesized that vFLIP might also activate this pathway. To evaluate this hypothesis, we transiently and stably transfected a vFLIP expression construct and performed several complementary assays to document that vFLIP activates the JNK/AP1 pathway and does so in a TRAF-dependent fashion. As vFLIP also activates the nuclear factor kappaB (NF-kappaB) signaling pathway and the NF-kappaB and JNK/AP1 pathways both modulate cellular interleukin-6 (cIL-6) expression, we postulated that vFLIP induces expression of this cytokine. We show that vFLIP induces cIL-6 expression and activates the cIL-6 promoter, and maximal activation of the cIL-6 promoter by vFLIP requires NF-kappaB and AP1 activation. In addition, vFLIP and latency-associated nuclear antigen (LANA), another KSHV-encoded latent protein, potentiate each other's ability to activate the cIL-6 promoter. Gene silencing experiments by RNA interference demonstrate that vFLIP in BCBL-1 endogenously infected primary effusion lymphoma (PEL) cells mediates JNK/AP1 activation and cIL-6 expression. Thus, we conclude that vFLIP, in addition to its known effects on NF-kappaB activation, also modulates the JNK/AP1 pathway and induces gene expression from the cIL-6 promoter in a JNK/AP1-dependent fashion. PMID- 12776189 TI - Functional characterization of IL-13 receptor alpha2 gene promoter: a critical role of the transcription factor STAT6 for regulated expression. AB - Interleukin (IL)-4 and IL-13 are two structurally and functionally related cytokines that have overlapping but also distinct biological activities. One of the components of the IL-13 receptor, the alpha2 chain (IL-13Ralpha2), has been reported to downregulate the cell responsiveness to IL-13, without affecting IL-4 signaling. Here, we report that TNFalpha synergizes with either IL-4 or IL-13 in inducing the IL-13Ralpha2 chain at both the mRNA and protein levels in the HaCaT human keratinocyte cell line. Further studies by 5'RACE identified as yet undescribed exonic sequences of the IL-13Ralpha2 5'UTR, provided evidence for the expression of alternatively spliced IL-13Ralpha2 transcripts and defined the transcription start of the IL-13Ralpha2 gene. A 1.5 kb region upstream of the first exon of the IL-13Ralpha2 gene displayed basal promoter activity when inserted in a reporter plasmid and transiently transfected in HaCaT cells. This promoter activity was further increased in response to IL-4 and IL-13. Furthermore, by electrophoretic mobility shift assay and site-directed mutagenesis, we showed that the IL-4/IL-13-induced promoter activity depended upon a positively acting STAT6 response element. Finally, TNFalpha was shown to potentiate IL-4/IL-13-induced IL-13Ralpha2 promoter activity when the same reporter construct was studied in stably but not in transiently transfected cells. These results suggest that the synergistic effect of TNFalpha on IL-4/IL 13-induced IL-13Ralpha2 expression is dependent upon chromatin re-modeling events. PMID- 12776190 TI - The mixed lineage leukemia fusion partner AF9 binds specific isoforms of the BCL 6 corepressor. AB - The mixed lineage leukemia (MLL) gene at chromosome band 11q23 is commonly involved in reciprocal translocations that are detected in acute leukemias. Evidence suggests that the resulting MLL fusion genes contribute to leukemogenesis. AF9 is a common MLL fusion partner in acute myeloid leukemia. The AF9 protein functions as a transcriptional activator in artificial reporter gene assays and a structurally related protein in yeast, ANC1/TFG3, is a component of the SWI/SNF complex. Apart from these observations, little is known about the biologic function of AF9 in mammals. We have found that a recently described transcriptional repressor, BCL-6 corepressor (BCoR), interacts with the carboxy terminus of AF9. The interaction of AF9 with BCoR has been confirmed by independent in vitro and in vivo protein-binding studies. The BCoR gene is expressed as several alternatively spliced transcripts. AF9 only binds BCoR isoforms that contain a unique 34 aa sequence located in the mid-portion of the protein. In artificial reporter gene assays, a BCoR isoform that binds AF9 efficiently suppresses AF9 transcriptional activity, while a nonbinding isoform does not. These results indicate that different isoforms of BCoR have unique biologic properties and that cell function may be partly determined by the different isoforms that are present within the cell. PMID- 12776191 TI - Loss of GFAP expression in high-grade astrocytomas does not contribute to tumor development or progression. AB - In astrocytic neoplasms, the number of cells expressing glial fibrillary acidic protein (GFAP) is inversely proportional to the extent of anaplasia. The loss of GFAP expression, the principal marker of astroglial cells, in these tumors has been proposed to constitute a step in their development and progression. To test this hypothesis, we crossed p53-negative (p53(-/-)) mice, which frequently develop astrocytomas after intrauterine exposure to ethylnitrosourea, with GFAP negative (GFAP(-/-)) mice or GFAP(+/+) controls. Brain tumors of glial origin were found in 12 of 35 GFAP(+/+) p53(-/-) mice (34%) and in 11 of 27 GFAP(-/-) p53(-/-) mice (41%). The two groups did not differ in the age at which tumors were detected or in tumor histology or progression. Thus, the loss of GFAP expression frequently seen in high-grade astrocytomas does not constitute a step in tumor development. Rather, it may represent the undifferentiated state of these cells. PMID- 12776192 TI - Peroxisome proliferator-activated receptor gamma is frequently downregulated in a diversity of sporadic nonmedullary thyroid carcinomas. AB - Peroxisome proliferator-activated receptor gamma (PPARgamma) has previously been implicated in the pathogenesis of follicular thyroid carcinoma (FTC), where a translocation with PAX8 has been reported in some 50% of tumors in three small series. The resultant fusion protein inhibits normal PPARgamma function by a dominant-negative mechanism. In a series of 19 FTCs, we identified this translocation in only two tumors (10.5%). However, microarray analysis and semiquantitative RT-PCR demonstrated greatly reduced PPARgamma expression in 13 of 17 (76%) nontranslocation tumors. Immunohistochemical analysis of 142 thyroid tumors showed a statistically significant reduction in PPARgamma immunoreactive protein, not only in FTCs but also in papillary thyroid carcinomas and Hurthle cell carcinomas. This suggests that while the overall frequency of the PAX8 PPARgamma translocation in FTCs may be lower than previously thought, functional downregulation of PPARgamma is a key event in multiple types of thyroid neoplasia and is a possible target for therapeutic intervention. PMID- 12776193 TI - ZNF198 protein, involved in rearrangement in myeloproliferative disease, forms complexes with the DNA repair-associated HHR6A/6B and RAD18 proteins. AB - A highly specific t(8;13)(p11;q12) translocation has been consistently identified in bone marrow cells from patients with an atypical myeloproliferative disease that is associated with peripheral blood eosinophila and T- or B-cell leukemias. In all patients analysed to date, the translocation event results in a chimeric gene in which the atypical zinc-finger domain of ZNF198 is fused to the N terminal end of the catalytic domain of the FGFR1 receptor tyrosine kinase. To understand more about the consequences of this rearrangement we have investigated the normal function of the ZNF198 gene. Using yeast two-hybrid analysis we identified HHR6 as a protein binding partner and confirmed this using immunoprecipitation studies. The ZNF198/FGFR1 fusion protein also binds to HHR6. We demonstrate here that the human RAD18 is also present in the ZNF198/HHR6 protein complex, although it does not coimmunoprecipitate with the fusion kinase. Cells expressing the fusion kinase gene show a marked increased sensitivity to UVB irradiation, suggesting that it acts in a dominant-negative way to affect DNA repair. These observations support the idea that ZNF198, through its interaction with HHR6 and RAD18, may be involved in the DNA repair process. PMID- 12776195 TI - Hypoxia attenuates the p53 response to cellular damage. AB - The tumour suppressor activity of p53 in vivo can be subject to pressure from the physiological stress of hypoxia and we report on the development of a cell system to define the p53-dependent stages in the adaptation of cells to hypoxia. p53(+/+) cells exposed to hypoxia exhibited a transient arrest in G2/M, but escaped from this checkpoint and entered a long-term G(0)/G(1) arrest. By contrast, isogenic p53-null cells exposed to hypoxic conditions exhibited a 6-10 fold higher level of apoptosis, suggesting that p53 acts as a survival factor under limiting oxygen concentrations. Surprisingly, hypoxia-dependent growth arrest in p53(+/+) cells did not result in either p21(WAF1) or HIF-1 protein stabilization, but rather promoted a significant decrease in Ser(392)-site phosphorylation at the CK2/FACT site. However, chemically induced anoxia induced Ser(392)-site phosphorylation as well as stabilization of both p53 and HIF-1 proteins. In contrast to hypoxia, 5-flourouracil (5-FU)-induced p53-dependent cell death correlated with enhanced Ser(392) phosphorylation of p53 and elevated p21(WAF1) protein levels. Hypoxia inhibited 5-FU-induced p53-dependent cell death and attenuated p53 phosphorylation at the ATM and CK2/FACT phosphorylation sites. Although anoxia activates the p53 response, hypoxia silences the p53 transactivation pathway and identifies a physiological signalling model to study mechanisms of p53 inactivation under hypoxic conditions. PMID- 12776194 TI - Overexpression of Kruppel-like factor 4 in the human colon cancer cell line RKO leads to reduced tumorigenecity. AB - Kruppel-like factor 4 (KLF4) is a zinc-finger-containing transcription factor, the expression of which is enriched in the postmitotic cells of the intestinal epithelium. KLF4 is a target gene of the tumor suppressor adenomatous polyposis coli (APC). We sought to determine the role of KLF4 in suppressing the tumorigenecity of RKO colon cancer cells, which do not express KLF4. We utilized an established system in RKO cells, in which an inducible promoter controls expression of KLF4. Four independent assays were used to assess the effects of KLF4 induction on tumor cells. We find that KLF4 overexpression reduces colony formation, cell migration and invasion, and in vivo tumorigenecity. The mechanism of action of KLF4 does not involve apoptosis. These findings, along with our previous findings that KLF4 induces G1/S arrest, suggest that KLF4 is a cell cycle checkpoint protein that can reduce tumorigenecity of colon cancer cells. PMID- 12776196 TI - A deletion mutant of heregulin increases the sensitivity of breast cancer cells to chemotherapy without promoting tumorigenicity. AB - Heregulin (HRG) is an activator of the erbB2-, erbB3- and erbB4-(erbB-2/3/4) signaling pathway. Transfection of full-length HRG cDNA into the estrogen (E2) dependent cell line MCF-7 promoted an invasive E2-independent phenotype, as well as persistent activation of the erbB-2/3/4 receptors. Moreover, HRG expression in MCF-7 cells renders the cells sensitive to the topoisomerase II inhibitor doxorubicin (Doxo). In an attempt to dissociate the tumorigenic effect of HRG from the sensitizing effect to chemotherapy, we constructed a structural deletion mutant of HRG. Transfection of the deletion mutant of HRG described in this study (HRG/M) into MCF-7 cells resulted in the dissociation of the tumor-promoting activity of HRG from the sensitization to Doxo, that is, although the cells did not become more aggressive or E2-independent they became more sensitive to Doxo. HRG/M was unable to autophosphorylate the erbB receptors and did not affect the level of MAPK phosphorylation. Furthermore, the intracellular localization of the protein was different from that of the full-length protein. Our data show that the HRG/M sequences are sufficient to sensitize MCF-7 cells to Doxo, and provide evidence that this sensitization is independent of erbB2 activation. PMID- 12776197 TI - Cyclin D1 is necessary but not sufficient for anchorage-independent growth of rat mammary tumor cells and is associated with resistance of the Copenhagen rat to mammary carcinogenesis. AB - To identify genes associated with the resistance of Copenhagen (Cop) rats to mammary carcinogenesis, we infused a retrovirus harboring v-Ha-ras directly into the main mammary ducts of resistant F1 rats from a cross between Cop and susceptible Wistar Furth (WF) rats. Adenocarcinomas formed in approximately 50% of infused glands. Cell lines derived from these tumors were clonal, but did not share a common viral integration site, suggesting that a high level of v-Ha-ras expression was able to overcome resistance in the F1 rats. Some of the cell lines were able to grow in soft agar, but a significant number did not display anchorage-independent growth. These growth characteristics were independent of v Ha-ras expression levels. The ability to grow in soft agar was associated with the size of tumors induced by injecting the cells into nude mice, and showed a striking positive association with the expression of cyclin D1. Furthermore, while resistance to anchorage-independent growth was fully overcome by transfection of cyclin D1 in some clones, in the others the effect was partial. A similar pattern of cyclin D1 upregulation and growth in soft agar was also observed when the cells were transfected with an active form of beta-catenin. Hybrid cells from the somatic fusion of an anchorage-dependent to an anchorage independent clone did not grow in soft agar. These results suggest that while a high expression level of cyclin D1 is necessary for anchorage-independent growth in all clones, it is not sufficient for full growth capacity in soft agar, raising the possibility that the loss of a tumor suppressor gene in the cell lines is required to fully confer anchorage-independent growth. Our anchorage dependent and -independent rat mammary tumor-derived cell lines may recapitulate the resistance and susceptibility of Cop and WF rats, respectively, to mammary carcinogenesis that could facilitate the identification of breast cancer susceptibility genes. PMID- 12776198 TI - MAL gene expression in esophageal cancer suppresses motility, invasion and tumorigenicity and enhances apoptosis through the Fas pathway. AB - We isolated the MAL (T-lymphocyte maturation associated protein) gene from differentially expressed products of esophageal epithelium relative to esophageal carcinoma tissues. The Mal protein has been demonstrated as being a component of the protein machinery for apical transport in epithelial polarized cells. In this study, we describe the reduced expression of MAL in all 39 cases of esophageal carcinoma tested and 60 other human carcinomas. MAL gene transcription was induced in three out of 13 esophageal carcinoma cell lines by treatment with the demethylating agent 5-aza-2'-deoxycytidine (DAC), and in nine additional cell lines by simultaneous treatment with trichostatin A, an inhibitor of deacetylation, and DAC. We established a stable MAL gene transfectant whose expression was regulated by subcutaneous doxycycline injection in nude mice. Tumor growth was suppressed in cells expressing TE3-MAL compared with TE3 parent cells or cells not expressing TE3-MAL with doxycycline injection (20 microg/body) (P<0.01). Additionally, the TE3-MAL transfectant cells exhibited decreased cellular motility, a G1/S transition block and increased levels of apoptosis, concomitant with increased expression of Fas receptor in vitro. The apoptotic staining in MAL-expressing tumors was confirmed by TUNEL assay. Therefore, we conclude that expression of MAL was frequently decreased or diminished in gastrointestinal tract cancers, and that Mal expression confers reduced tumorigenicity in vivo to tumor TE3 cells through the induction of apoptosis via the Fas signaling pathway. PMID- 12776199 TI - LOH of PTPRJ occurs early in colorectal cancer and is associated with chromosomal loss of 18q12-21. AB - Recently, the gene PTPRJ (protein tyrosine phosphatase receptor type J) was identified as the candidate gene for the mouse colon cancer susceptibility locus Scc1. Its human homologue PTPRJ is frequently deleted in several cancer types, including colorectal cancer. To elucidate the role of PTPRJ loss in different stages of colorectal cancer and in its pathways of progression, we expanded the previously published comparative genomic hybridization results with novel data on loss of heterozygosity (LOH) at the PTPRJ locus. We identified a strong association between the LOH of PTPRJ and the loss of chromosomal region 18q12-21 (P=0.009). This observation is specific for progressed colorectal adenomas, suggesting that an interaction between LOH of PTPRJ and loss of 18q12-21 may be involved in the development of a more progressed form of adenomas. PMID- 12776200 TI - Effects of methylation on expression of TMS1/ASC in human breast cancer cells. AB - Gene silencing associated with aberrant methylation of promoter region CpG islands is one mechanism in which tumor suppressor genes are inactivated in human cancers. Recently, we identified a novel gene, Target of Methylation-associated Silencing-1 (TMS1) (also called ASC), which is aberrantly methylated and silenced in human breast cancers. To further investigate the mechanism of TMS1 silencing, we defined the transcription initiation site and detailed the DNA methylation pattern of the TMS1 CpG island in normal breast epithelial cells, breast cancer cell lines, and primary tumors. We find that in normal cells, the TMS1 CpG island lies within a 1.2 kb unmethylated domain, the 5' boundary of which is in close proximity to the transcription initiation site. In breast cancer cell lines, this boundary appeared to be unstable in that methylation tended to accumulate in the 5' end of the CpG island relative to normal epithelial cells. In contrast, the 3' methylation boundary remained intact. Gene silencing was not correlated with the methylation of specific CpG sites nor the inability to transactivate the TMS1 promoter, but was correlated with the percentage of alleles in the population exhibiting dense methylation across the entire CpG island. Using 5-aza deoxycytidine to reactivate TMS1 in methylated and silent cell lines, and a cassette methylation strategy to determine the impact of methylation on different parts of the promoter, we find that demethylation of a small region immediately surrounding the transcription start site is critical to TMS1 expression. Our data support the idea that gene silencing and dense methylation are tightly coupled events that affect individual chromosomal copies of TMS1 in an all-or-none manner. The transition to this stably repressed state may be facilitated by spreading of methylation into the proximal promoter and direct effects of methylation on TMS1 transcription. PMID- 12776201 TI - Genetic analysis identifies putative tumor suppressor sites at 2q35-q36.1 and 2q36.3-q37.1 involved in cervical cancer progression. AB - We performed comparative genomic hybridization (CGH) and high-resolution deletion mapping of the long arm of chromosome 2 (2q) in invasive cervical carcinoma (CC). The CGH analyses on 52 CCs identified genetic losses at 2q33-q36, gain of 3q26 q29, and frequent chromosomal amplifications. Characterization of 2q deletions by loss of heterozygosity (LOH) in 60 primary tumors identified two sites of minimal deleted regions at 2q35-q36.1 and 2q36.3-q37.1. To delineate the stage at which these genetic alterations occur in CC progression, we analysed 33 cervical intraepithelial neoplasia (CIN) for LOH. We found that 89% of high-grade (CINII and CINIII) and 40% of low-grade (CINI) CINs exhibited LOH at 2q. To identify the target tumor suppressor gene (TSG), we performed an extensive genetic and epigenetic analyses of a number of candidate genes mapped to the deleted regions. We did not find inactivating mutations in CASP10, BARD1, XRCC5, or PPP1R7 genes mapped to the deleted regions. However, we did find evidence of downregulated gene expression in CFLAR, CASP10 and PPP1R7 in CC cell lines. We also found reactivated gene expression in CC cell lines in vitro after exposure to demethylating and histone deacetylase (HDAC) inhibiting agents. Thus, these data identify frequent chromosomal amplifications in CC, and sites of TSGs at 2q35 q36.1 and 2q36.3-q37.1 that are critical in CC development. PMID- 12776202 TI - Differential molecular profiling between skin carcinomas reveals four newly reported genes potentially implicated in squamous cell carcinoma development. AB - Basal cell carcinoma (BCC) and squamous cell carcinoma (SCC) are skin tumors with different invasive potential. In this work, we analysed mRNA differential expression between seven BCC and five SCC and their normal skin counterparts using 1176 cDNA macroarrays and verification by RT-PCR to identify genes modulated in each tumor type. We identified 37 genes commonly modulated in both tumors and four genes specifically modulated in SCC. Among these latter RhoC and EMMPRIN genes seem to be of particular interest and could participate in SCC aggressivity. PMID- 12776203 TI - MBD1, MBD2 and CGBP genes at chromosome 18q21 are infrequently mutated in human colon and lung cancers. AB - The genes MBD1 and MBD2 encode methyl-CpG binding proteins that suppress transcription from methylated promoters. In contrast, CGBP encodes a protein that binds promoters containing unmethylated CpG and stimulates transcription. All three are located on human chromosome 18q21, a region of frequent loss of heterozygosity in several cancers. These genes therefore represent candidate tumour suppressor genes, whose loss of function could affect the normal regulation of gene expression, whether by lack of complete suppression of genes normally silenced (via loss of MBD1 and MBD2) or by some loss of activation of genes normally expressed (via loss of CGBP), either way contributing to the tumorigenic phenotype. We have confirmed by fluorescent in situ hybridization that MBD1 and MBD2 bracket the DCC locus giving a gene order of MBD1/CGBP-DCC 5' DCC 3'-MBD2. Mutation analyses by single-stranded conformation polymorphism in colon and lung cancer cell lines and primary tumours revealed a small number of mutations, suggesting only a limited role of these genes in human tumorigenesis. PMID- 12776204 TI - TREMs in the immune system and beyond. AB - Triggering receptors expressed by myeloid cells (TREMs) belong to a rapidly expanding family of receptors that include activating and inhibitory isoforms encoded by a gene cluster linked to the MHC. TREM1 and TREM2 activate myeloid cells by signalling through the adaptor protein DAP12. TREM1 triggers phagocyte secretion of pro-inflammatory chemokines and cytokines, amplifying the inflammation that is induced by bacteria and fungi. TREM2 activates monocyte derived dendritic cells and regulates osteoclast development. Remarkably, TREM2 deficiency leads to a severe disease that is characterized by bone cysts and demyelination of the central nervous system, which results in dementia, implying that the function of TREM2 extends beyond the immune system. PMID- 12776205 TI - The TIM gene family: emerging roles in immunity and disease. AB - The search for cell-surface markers that can distinguish T helper 1 (T(H)1) cells from T(H)2 cells has led to the identification of a new gene family, encoding the T-cell immunoglobulin mucin (TIM) proteins, some of which are differentially expressed by T(H)1 and T(H)2 cells. The role of the TIM-family proteins in immune regulation is just beginning to emerge. Here, we describe the various TIM-family members in mice and humans, and discuss the genetic and functional evidence for their role in regulating autoimmune and allergic diseases. PMID- 12776206 TI - A forward-genetic approach for analysis of the immune system. AB - The completion of the genome sequences of both humans and mice challenges biologists to determine gene function on a vast, whole-organism scale. Both phenotype-based ('forward') and gene-based ('reverse') strategies are being developed to approach this issue. Forward-genetic approaches, however, provide the unique ability of assigning function to genes in an unbiased, global manner that is independent of previous assumptions about gene function. In this article, we compare various genetic technologies for their potential role in dissecting immune-system development and function, with particular emphasis on the worldwide efforts that use chemical mutagenesis as a forward-genetic strategy. PMID- 12776207 TI - Lysosomal cysteine proteases regulate antigen presentation. AB - Antigen presentation by both classical MHC class II molecules and the non classical MHC class I-like molecule CD1D requires their entry into the endosomal/lysosomal compartment. Lysosomal cysteine proteases constitute an important subset of the enzymes that are present in this compartment and, here, we discuss the role of these proteases in regulating antigen presentation by both MHC class II and CD1D molecules. PMID- 12776208 TI - Transcriptional analysis of targets in multiple sclerosis. AB - Large-scale analyses of messenger RNA transcripts and autoantibody responses, taken from the actual sites of disease, provide us with an unprecedented view of the complexity of autoimmunity. Despite an appreciation of the large number of pathways and pathological processes that are involved in these diseases, a few practical targets and several new strategies have emerged from these studies. This review focuses on multiple sclerosis and on the approaches that are being used to identify new targets that might be manipulated to control this disease. PMID- 12776209 TI - Immune responses to RNA-virus infections of the CNS. AB - A successful outcome for the host of virus infection of the central nervous system (CNS) requires the elimination of the virus without damage to essential non-renewable cells, such as neurons. As a result, inflammatory responses must be tightly controlled, and many unique mechanisms seem to contribute to this control. In addition to being important causes of human disease, RNA viruses that infect the CNS provide useful models in which to study immune responses in the CNS. Recent work has shown the importance of innate immune responses in the CNS in controlling virus infection. And advances have been made in assessing the relative roles of cytotoxic T cells, antibodies and cytokines in the clearance of viruses from neurons, glial cells and meningeal cells. PMID- 12776210 TI - Therapeutic vaccines against HIV need international partnerships. AB - The successes of anti-retroviral treatments against HIV are limited today by the cost and toxicity of lifelong administration. An innovative therapeutic strategy has been proposed to boost the immune system of infected patients with HIV vaccines and to help limit the use of anti-retroviral treatments. This perspective article reviews the crucial questions raised by such a strategy and the main international efforts that are already set up to provide rapid answers- in particular, a not-for-profit international network that is dedicated to the development of therapeutic immunization programmes against HIV. PMID- 12776211 TI - Sequence-based cancer genomics: progress, lessons and opportunities. AB - Technologies that provide a genome-wide view offer an unprecedented opportunity to scrutinize the molecular biology of the cancer cell. The information that is derived from these technologies is well suited to the development of public databases of alterations in the cancer genome and its expression. Here, we describe the synergistic efforts of research programmes in Brazil, the United Kingdom and the United States towards building integrated databases that are widely accessible to the research community, to enable basic and applied applications in cancer research. PMID- 12776212 TI - The art and design of genetic screens: Escherichia coli. AB - This article summarizes the general principles of selections and screens in Escherichia coli. The focus is on the lac operon, owing to its inherent simplicity and versatility. Examples of different strategies for mutagenesis and mutant discovery are described. In particular, the usefulness and effectiveness of simple colour-based screens are illustrated. The power of lac genetics can be applied to almost any bacterial system with gene fusions that hook any gene of interest to lacZ, which is the structural gene that encodes beta-galactosidase. The diversity of biological processes that can be studied with lac genetics is remarkable and includes DNA metabolism, gene regulation and signal transduction, protein localization and folding, and even electron transport. PMID- 12776213 TI - Exploiting genomics, genetics and chemistry to combat antibiotic resistance. AB - To address the worsening problem of antibiotic-resistant bacteria there is an urgent need to develop new antibiotics. Comparative genomics and molecular genetics are being applied to produce lists of essential new targets for compound screening programmes. Combinatorial chemistry and structural biology are being applied to rapidly explore and optimize the interactions between lead compounds and their biological targets. Several compounds that have been identified from target-based screens are now in development, but technical and economic constraints might result in a trickle, rather than a flood, of new antibiotics onto the market in the near future. PMID- 12776214 TI - Genetic strategies for antibacterial drug discovery. AB - The availability of genome sequences is revolutionizing the field of microbiology. Genetic methods are being modified to facilitate rapid analysis at a genome-wide level and are blossoming for human pathogens that were previously considered intractable. This revolution coincided with a growing concern about the emergence of microbial drug resistance, compelling the pharmaceutical industry to search for new antimicrobial agents. The availability of the new technologies, combined with many genetic strategies, has changed the way that researchers approach antibacterial drug discovery. PMID- 12776215 TI - Evolution experiments with microorganisms: the dynamics and genetic bases of adaptation. AB - Microorganisms have been mutating and evolving on Earth for billions of years. Now, a field of research has developed around the idea of using microorganisms to study evolution in action. Controlled and replicated experiments are using viruses, bacteria and yeast to investigate how their genomes and phenotypic properties evolve over hundreds and even thousands of generations. Here, we examine the dynamics of evolutionary adaptation, the genetic bases of adaptation, tradeoffs and the environmental specificity of adaptation, the origin and evolutionary consequences of mutators, and the process of drift decay in very small populations. PMID- 12776216 TI - The future of bacteriophage biology. AB - After an illustrious history as one of the primary tools that established the foundations of molecular biology, bacteriophage research is now undergoing a renaissance in which the primary focus is on the phages themselves rather than the molecular mechanisms that they explain. Studies of the evolution of phages and their role in natural ecosystems are flourishing. Practical questions, such as how to use phages to combat human diseases that are caused by bacteria, how to eradicate phage pests in the food industry and what role they have in the causation of human diseases, are receiving increased attention. Phages are also useful in the deeper exploration of basic molecular and biophysical questions. PMID- 12776217 TI - Modelling genitourinary defects in mice: an emerging genetic and developmental system. AB - The rising incidence of genitourinary (GU) defects among newborns establishes the need and opportunity to focus research efforts on the amelioration of this growing public-health concern. Sadly, our inability to explain the causes of GU defects can be directly attributed to our lack of understanding of GU gene function. Recently, mouse models have been used to provide new insights into the mechanisms that underlie congenital GU malformations. PMID- 12776218 TI - Predicting therapeutic value in the lead optimization phase of drug discovery. AB - Recombinant and natural cellular assays for human G-protein-coupled receptors are used to optimize initial lead molecules obtained from screening. Although the activity of these molecules can be assessed on human genotype receptors, there is increasing evidence that cells impose a phenotypic selectivity to molecules in various cellular backgrounds. This opens the possibility of dissimulations between activity seen in lead optimization assays and the intended therapeutic value in humans. This review discusses the mechanisms by which cells can impose phenotypic selectivity on molecules and approaches to reduce this practical problem for drug discovery. PMID- 12776219 TI - The impact of drug-induced QT interval prolongation on drug discovery and development. AB - During the past decade, a number of non-cardiovascular drugs have had their label revised or have been withdrawn from the market because of unexpected post marketing reports of sudden cardiac death associated with a prolongation of the QT interval, and an increased propensity to develop a ventricular tachyarrhythmia called Torsades de Pointes. Although a direct link between QT interval prolongation and arrhythmogenesis is still unclear, QT prolongation is now the subject of increased regulatory review and is considered a significant risk factor for predicting human safety of New Chemical Entities. Consequently, pharmaceutical companies are striving to improve the drug discovery and development process to identify, as early as possible, the risk of novel agents, or their metabolites, of causing QT interval prolongation and to make appropriate go/no-go decisions or modify their development programme accordingly. PMID- 12776220 TI - Ophthalmic drug discovery. AB - Millions of people suffer from a wide variety of ocular diseases, many of which lead to irreversible blindness. The leading causes of irreversible blindness in the elderly--age-related macular degeneration and glaucoma--will continue to effect more individuals as the worldwide population continues to age. Although there are therapies for treating glaucoma, as well as ongoing clinical trials of treatments for age-related macular degeneration, there still is a great need for more efficacious treatments that halt or even reverse ocular diseases. The eye has special attributes that allow local drug delivery and non-invasive clinical assessment of disease, but it is also a highly complex and unique organ, which makes understanding disease pathogenesis and ocular drug discovery challenging. As we learn more about the cellular mechanisms involved in age-related macular degeneration and glaucoma, potentially, new drug targets will emerge. This review provides insight into some of the new approaches to therapy. PMID- 12776221 TI - Neurotrophic factors as novel therapeutics for neuropathic pain. AB - Neuropathic pain is a chronic condition that is caused by injury to the nervous system. Unlike acute pain, which is protective, neuropathic pain persists and serves no useful purpose, and severely affects quality of life. However, present therapies have modest efficacy in most patients, are palliative rather than curative, and their side effects represent significant limitations. Tremendous progress has been made over the past decade in our understanding of the biology of pain sensory neurons. The recent discovery that neurotrophic factors play an important role in neuropathic pain indicates that these pathways could serve as novel intervention points for therapy. Moreover, neurotrophic factors have the potential to address the underlying pathophysiology of neuropathic pain, thereby halting or reversing the disease process. PMID- 12776222 TI - Therapeutic strategies for rheumatoid arthritis. AB - Recent years have seen considerable advances in our understanding of both the clinical and basic-research aspects of rheumatoid arthritis. Clinical progress has come from a better recognition of the natural history of the disease, the development and validation of outcome measures for clinical trials and, consequently, innovative trial designs. In parallel, basic research has provided clues to the pathogenic events underlying rheumatoid arthritis, and advances in biotechnology have facilitated the development of new classes of therapeutics. Here, we summarize the fruits of these advances: innovative approaches to the use of existing, traditional disease-modifying antirheumatic drugs; novel agents approved very recently; and further avenues that are presently under investigation or which are of more distant promise. PMID- 12776223 TI - The prospect for bacteriophage therapy in Western medicine. AB - Bacteriophage (phage) have been used for clinical applications since their initial discovery at the beginning of the twentieth century. However, they have never been subjected to the scrutiny--in terms of the determination of efficacy and pharmacokinetics of therapeutic agents--that is required in countries that enforce certification for marketed pharmaceuticals. There are a number of historical reasons for this deficiency, including the overshadowing discovery of the antibiotics. Nevertheless, present efforts to develop phage into reliable antibacterial agents have been substantially enhanced by knowledge gained concerning the genetics and physiology of phage in molecular detail during the past 50 years. Such efforts will be of importance given the emergence of antibiotic-resistant bacteria. PMID- 12776224 TI - Investigation of the cerebral energy status in patients with glutaric aciduria type I by 31P magnetic resonance spectroscopy. AB - In vivo phosphorus magnetic resonance spectroscopy (MRS) was used to investigate markers of the cerebral energy status in two patients with glutaric aciduria type I (GA-I). Besides an increased concentration of phosphomonoesters in one patient, no other significant alterations from controls were found. This might indicate increased resynthesis of dendritic processes secondary to preceding metabolic crises. In contrast to previous cell-culture studies, no cerebral depletion of phosphocreatine (PCr) was observed. In conclusion, a severe global and permanent depletion of cerebral energy supplies must be ruled out. The benefit of a permanent creatine substitution to stabilize mitochondrial energy metabolism seems thus questionable. However, as MRS was performed during stable clinical conditions, the possibility of a PCr decrease during acute metabolic crises cannot be assessed. PMID- 12776225 TI - General movements detect early signs of hemiplegia in term infants with neonatal cerebral infarction. AB - Background. Studies have reported that infants with hemiplegia of congenital origin may have a period between birth and up to 12 months when clinical signs of hemiplegia are not evident. The aim of this study was to establish whether the assessment of general movements (GMs) may help in the earlier detection of signs of hemiplegia. Subjects and Methods. Eleven infants with cerebral infarction on brain MRI, and eleven normal controls were enrolled in the study. Quality of GMs was assessed from videotapes between 3 and 6 weeks and between 9 and 16 weeks. Neurological outcome was evaluated at least at two years. Results. Seven of the 11 infants had an assessment performed between 3 and 6 weeks: abnormal GMs were observed in all the infants who developed hemiplegia, but one child had abnormal GMs and a normal outcome. All 11 infants had a scorable assessment between 9 and 16 weeks. In all a specific type of GMs, fidgety movements (FMs), were predictive of neurological outcome. The presence of early asymmetries at both 3 - 6 and 9 - 16 weeks was also significantly associated with later signs of hemiplegia. Conclusions. The assessment of GMs after the neonatal period appears to be very useful in the early identification of hemiplegia in infants with cerebral infarction. Whilst the prediction of hemiplegia should be possible from early neonatal MRI brain scans, this facility is not always available. Observation of GMs is a bedside clinical approach that allows confirmation of early prediction from MRI, early rehabilitation if needed and reassurance that neurological outcome will be good where that is appropriate. PMID- 12776226 TI - Follow-up of children with cerebral palsy after selective posterior rhizotomy with intensive physiotherapy or physiotherapy alone. AB - In all 21 children with spastic cerebral palsy (CP) underwent surgery involving selective posterior rhizotomy (SPR), followed by six months intensive physiotherapy (PT). Neurological and physiotherapeutic assessments were made one, three and five years after the operation. The children undergoing surgery were compared to 21 comparison children who took part in a regular physiotherapy programme during the same time period. At the preoperative assessment, the children undergoing surgery were similar to the comparative children in terms of age, sex, type of CP, spasticity of the legs and mean functional scores. The children were selected for SPR on the basis of more than half a year's arrest of motor development, which was the only significant difference to the comparative group. Motor function was measured using two different methods, the Illinois-St Louis Scale and the Gross Motor Functional Classification System (GMFC). Both groups experienced steady development during the five-year follow-up period and no significant differences were observed in the mean functional scores between the groups. We conclude that this comparative study, like most controlled studies, failed to demonstrate any additional effect of SPR on motor development of children with spastic CP. Nevertheless, SPR may contribute to a resumption of motor development in children with arrested motor development despite vigorous conservative therapy. SPR is therefore justified as treatment in selected cases. PMID- 12776227 TI - Diffusion-weighted MRI in severe neonatal hypoxic ischaemia: the white cerebrum. AB - Presently, prognosis of infants with perinatal hypoxia-ischaemia is estimated using the Sarnat scale, which combines clinical evaluation and electroencephalography, in combination with magnetic resonance imaging (MRI) and or evoked potentials. While the value of conventional MRI is limited during the first week of life, diffusion-weighted MRI demonstrates effects of acute cerebral ischaemia within hours of onset. However, the exact value of diffusion MRI in the prognosis of infants with hypoxia-ischaemia has to be established in larger follow-up studies. In this report we describe 5 term (post-conceptional age 40 1/7 to 41 2/7 week) neonates with severe hypoxia-ischaemia and a characteristic pattern of diffusion changes. T 1 -weighted images showed a hyperintense cortical signal in only one case and extensive hyperintensity in the basal nuclei in all 5 cases. T 2 -weighted images showed nearly complete loss of cortical delineation in three cases. Increased signal on diffusion-weighted images was seen throughout all cortical and subcortical areas while the cerebellum remained normal. This pattern, which we refer to as the "white cerebrum", is most readily apparent on coronal images. The apparent diffusion coefficient (ADC) was calculated and compared to that of four control infants. In the cortex ADC values were lowered (0.70 +/- 0.17 micro m 2/msec [mean +/- standard deviation (SD)]; controls [n = 4]: 1.18 +/- 0.02 micro m 2/msec) as compared to values of ADC in the cerebellum (1.31 +/- 0.06 micro m 2/msec [mean +/- SD]; controls [n = 4]: 1.25 +/- 0.06 micro m 2/msec). All infants died in the perinatal period. In summary, the "white cerebrum" on diffusion-weighted MRI indicates severe neonatal hypoxia-ischaemia and is the counterpart of the white cerebellum on CT. PMID- 12776228 TI - Methylphenidate down-regulates the dopamine receptor and transporter system in children with attention deficit hyperkinetic disorder (ADHD). AB - Adults suffering from Attention Deficit Hyperactivity Disorder (ADHD) are known to have disturbed central dopaminergic transmission. With Single Photon Emission Computed Tomography (SPECT) we studied brain dopamine transporter and receptor activity in six boys with ADHD. Three months after initiation of treatment with methylphenidate we found a down-regulation of the post-synaptic dopamine receptor with a maximum of 20 % and a down-regulation of the dopamine transporter with a maximum of 74.7 % in the striatal system. This corresponded to a positive clinical response evaluated by neuropsychological questionnaires and tests. We suggest that dopamine transporter imaging by SPECT might be used to monitor psychostimulant treatment in children suffering from ADHD. PMID- 12776230 TI - Leigh syndrome with cytochrome-c oxidase deficiency and a single T insertion nt 5537 in the mitochondrial tRNATrp gene. AB - We report a nine-year-old boy with the features of Leigh syndrome (LS) and a severe cytochrome-c oxidase (COX) deficiency with a single thymidine insertion at nucleotide position 5537 (T 5537i) in the tRNA Trp gene of mitochondrial DNA. During infancy the boy was irritable and hypotonus was noticed. Early motor development was delayed, although mental development seemed normal until eight months of age. Early neurological signs were nystagmus, hypertonus and optic atrophy. Severe seizures and mental retardation developed subsequently. Major findings on neuroradiological investigation were from the brainstem, thalami and white matter compatible with LS. Spectrophotometric analysis of skeletal muscle mitochondria showed a profound COX deficiency and a marked complex I deficiency. Enzyme-histochemical analysis showed reduced COX activity in the majority of the muscle fibres. There were no ragged red fibres. The T 5537i mutation was found in a high proportion (> 95 %) in blood, liver and muscle tissue of the patient and in blood of the patient's mother (81 %). This mutation has previously been described in one family in which one child had a very high proportion of the T 5537i mutation and clinical features of LS. We conclude that, although mtDNA mutations are considered to be rare in LS with COX deficiency, the T 5537i mutation should be screened for in cases of LS with COX deficiency when SURF1 gene mutations have been excluded, especially when complex I activity is also decreased. PMID- 12776229 TI - Longitudinal study of auditory brainstem response in leigh syndrome. AB - To assess the utility of auditory brainstem response (ABR) in diagnosing brainstem changes in patients with Leigh syndrome (LS), we performed a longitudinal study of five patients with LS using both ABR and neuroimaging techniques (CT and MRI). The brainstem components of the initial ABRs we performed on the patients were abnormal in all five patients. In four of the patients, these abnormal findings preceded any clinical signs of brainstem impairment. Improvements in clinical findings were reflected in improvements in ABR findings in three patients. In one of these three patients, improvements in clinical findings were also reflected in improvements in MRI findings. In the other two patients, MRI findings showed no improvements, despite the improvements in clinical findings. In two of our patients, ABR clearly revealed functional improvements in the brainstem which were not revealed by MRI. Therefore, we conclude that ABR is an essential diagnostic technique for patients with LS. PMID- 12776231 TI - Occipito-temporal polymicrogyria and subclinical muscular dystrophy. AB - We report a two-year-old Caucasian boy who had neonatal seizures and was found to have bilateral occipito-temporal polymicrogyria on neonatal brain MRI. The child had no additional neurological abnormality other than the neonatal seizures, but serum CK was found to be elevated (5 - 7 times normal values) and the muscle biopsy showed evidence of early muscular dystrophy. Detailed protein and genetic studies did not allow the identification of a known form of muscular dystrophy. The boy has been followed regularly and he currently has mild global developmental delay but no clinical signs of muscle involvement. The association of polymicrogyria and muscular dystrophy is known to occur in Fukuyama and Walker Warburg muscular dystrophies, in muscle-eye-brain disease and in some patients with merosin deficient CMD. However the absence of weakness and of eye involvement, the normal expression of merosin and alpha dystroglycan and the pattern of brain involvement make it very unlikely that the child is affected by one of these forms. As the pattern of brain involvement and the muscle pathology is not typical of one of the forms of neuronal migration disorders secondary to a known gene defect, we suspect that the combination of muscle and brain involvement found in this child is not coincidental. Our findings suggest that serum CK should be determined in children with undiagnosed polymicrogyria, even in the absence of weakness. This may lead to an expansion of our understanding of muscle dystrophies and cortical dysplasias. PMID- 12776232 TI - Ultrasound findings in follow-up investigations in a case of aspartoacylase deficiency (canavan disease). AB - Aspartoacylase deficiency is a neurodegenerative disease which typically starts in the first months of life with muscular hypotonia and developmental standstill. One of the first diagnostic procedures in this situation is an ultrasound of the brain. There is little information available about sonographic changes in Canavan disease. We present for the first time an ultrasound follow-up in a proven case of aspartoacylase deficiency from 3 weeks to 22 months. High echogenicity of the white matter was present in the neonatal period. Additional sonographic phenomena resulting in a characteristic pattern were shown in further investigations. The distinctive sonomorphology is compared to a few other cases in the literature. The correlation to the neuropathological course of the white matter changes is discussed. Recognition of the sonographic features in addition to the clinical presentation may contribute to an effective biochemical work-up. PMID- 12776233 TI - Acute leukoencephalopathy after inhalation of a single dose of heroin. AB - We describe extended and repeat magnetic resonance (MR) examinations in the case of a 16-year-old male who developed acute left-sided sensorimotor hemiplegia after a single dose of inhaled heroin. MRI revealed symmetrical hyperintense signals in T 2 -weighted images and massive diffusion disorders in the diffusion weighted images predominantly in parieto-occipital subcortical white matter and both ventral globi pallidi with preservation of U fibers and no brain oedema. MR spectroscopy data were compatible with combined hypoxic and mitochondrial damage resulting in axonal injury without demyelination. Normal values and variations had been obtained from spectra of five age-matched subjects. This is the first reported MR follow-up study of leukoencephalopathy occurring acutely after a first inhaled dose of heroin. We postulate that toxic spongiform leukoencephalopathy in heroin addicts may be the outcome of a complex mechanism directly triggered by heroin and causing mitochondrial as well as hypoxic injury in specific and limited areas of white matter. PMID- 12776234 TI - Treatment with Sulthiame (Ospolot) in benign partial epilepsy of childhood and related syndromes: an open clinical and EEG study. AB - The effect of Sulthiame on the EEG and on clinical seizures was evaluated in an open uncontrolled study in 25 children with focal sharp waves on the EEG (FSW). 16 children had typical benign partial epilepsy with rolandic spikes (BPERS), 5 children with atypical forms and 4 children with no clinical seizures but cognitive disturbances possibly related to the FSW. The effect of Sulthiame in suppressing the EEG discharges was evaluated on the waking and sleep EEG before introduction of the drug, and at 3 - 6 months, 6 to 12 months and beyond while under therapy. The children were followed clinically for one to several years. The EEG discharges disappeared or decreased under Sulthiame in 13/21 cases at 3 to 6 months but reappeared in 3/13 cases beyond this period. No case had a worsening of the EEG or of clinical seizures under Sulthiame, and no cognitive stagnation was noted. Our data confirm the good tolerance and positive effects on the EEG and justify systematic trials of this drug in the partial "functional" epilepsies, especially when negative cognitive consequences of the epileptic discharges are suspected. PMID- 12776235 TI - Antibodies against muscle-specific kinase in juvenile myasthenia gravis. PMID- 12776236 TI - Training in pediatric surgery--a comparison of 24 countries in Europe and other countries around the world. AB - What is the training in pediatric surgery like, how is pediatric surgery defined in other countries and what are the possibilities of quality control for training in pediatric surgery? The results of an inquiry together with information about training in pediatric surgery in 24 countries are summarised to show the different possibilities of organising training in pediatric surgery inside and outside Europe. The number of trainees in pediatric surgery (ranging from 0 to 339 trainees per country), the density of pediatric surgeons (ranging from 424 to 35 714 live births/year per pediatric surgeon) and the number of Centres of Pediatric Surgery (ranging from 4167 to 65,000 live births or 450,000 to 5,300,000 inhabitants per Centre) varies a lot in the different countries. Countries with a higher density of pediatric surgeons often also have a higher number of trainees irrespective of the birth rate and the number of inhabitants within the country, indicating a good infrastructure for pediatric surgery. In 87.5 % of the countries pediatric surgery is recognised as a specialty. The mean duration of training is about 6 years and 3 months, excluding the countries where it is necessary to become a fully accredited general surgeon before doing pediatric surgery. The mean duration of stay in general surgery is 2 years and 10 months. In one third of the countries it is obligatory to spend some time (3 to 12 months) in pediatrics. An elective or a compulsory period of time (1 - 6 months) in other specialties such as orthopedic surgery, plastic and reconstructive surgery, thoracic surgery and urology exists in 41.7 % of the nations. The most common subspecialties within pediatric surgery practised during the training by the resident are abdominal surgery, thoracic surgery, oncological surgery, head and neck surgery and urology. Possible means of control to guarantee a high quality of training include a defined time of stay in each subspecialty, the number of operations done by the trainee or the number of cases treated by the resident in the respective subspecialty and the distinction of different levels of surgery done by the trainee during his residence. In many cases the type of control is not specified. PMID- 12776237 TI - Experience with a new concept to lower non-infectious complications in infants with programmable shunts. AB - In neonates and infants less than 1 year of age who are treated with a ventriculo peritoneal shunt, non-infectious complications are almost as frequent and dangerous as infectious complications. While the incidence of infections can be reduced, using perioperative antibiotics, special surgical techniques and postoperative care non-infectious complications such as wound break-down, cerebrospinal fluid (CSF) fistula and subcutaneous CSF collection are preventable, but seem difficult to manage, especially in the group of patients at a susceptible age. The authors present their experience with the programmable Hakim valve in 40 neonates and infants less than 1 year of age, who were treated with de novo implantation of a ventriculo-peritoneal shunt due to various pathologies. The uneventful wound healing during the first weeks after shunt implantation, avoiding the above mentioned non-infectious complications, was supported by initial, temporary overdrainage and readjustment of the programmable valve after completed wound healing. All patients tolerated this procedure well and showed no pathological signs or symptoms of overdrainage like premature closure of cranial sutures, clinical low-pressure syndrome, slit ventricle syndrome (SVS), subdural fluid collection or brain collapse during an average follow-up period of 2.6 years (2 - 65 months). Infectious and other mechanical, non-infectious complications were analysed as well during the follow-up period. These results suggest that an initial, temporary overdrainage in infants and neonates with shunted hydrocephalus may contribute to further lower the incidence of non-infectious complications like wound break-down, CSF-fistula or subcutaneous CSF accumulation, without negative side effects. This technique could be a valuable option in the regimen of shunt-treatment of this age group in order to optimise the overall success rate and lower the general complication rate. PMID- 12776238 TI - Evaluation of cerebral vascular reserve by single photon emission tomography in children with congenital hydrocephalus. AB - BACKGROUND/AIMS: Single photon emission tomography scan (SPECT) is used to measure cerebral blood flow (CBF) and also cerebral vascular reserve (CVR) after intravenous acetazolamide (ACZ) challenge. This prospective study was designed to test the utility of CVR in congenital hydrocephalus (CHC). METHODS: Patients (n = 21, mean age 6.2 m) with CHC underwent a baseline ultrasonography (US), retinal fundus examination (RF), neurological score (NS), and SPECT to evaluate CVR after intravenous ACZ. Monthly US, RF, NS, CBF, and CVR were evaluated. RESULTS: Ten patients showed good CVR and also had stable NS and RF despite mild to moderate ventriculomegaly and were followed up on conservative treatment. None of these patients in a mean follow-up period of 9.8 months showed deterioration. Eleven patients, who had poor CVR initially, were also treated conservatively. Over a period of 4 months, 8 patients showed a fall in CVR to < 30 % of baseline and a fall in NS despite RF and US remaining the same and hence were operated. Five of these operated cases showed an improvement in CVR and NS over a mean follow-up period of 4 months and the remaining 3 patients failed to show improvement in CVR and NS. CONCLUSIONS: In CHC, CVR measured by ACZ challenge test is a sensitive parameter as alterations in CVR occur prior to changes in other conventional parameters such as ventriculomegaly and RF. Patients with normal CVR at presentation are unlikely to deteriorate and can safely be managed conservatively. Patients who show a deterioration in CVR should be considered for prompt surgery. PMID- 12776239 TI - Lower lip repair in Van der Woude syndrome. AB - Van der Woude syndrome is an unusual congenital malformation. It combines lower lip sinuses with a cleft lip and/or palate. The asymmetry appears more pronounced due to lower lip hypotonia and procheilia, which are more unaesthetic than the fistulae themselves. The malformation of the orbicularis oris muscle in its medial part induces the hypotonic aspect of the lower lip, which contrasts with the retracted scarred upper lip after the cleft has been repaired. It is also responsible for a real bilabial asymmetry. For many years, the most common surgical treatment was simple excision of the sinuses. Bad functional repair results prompted the authors to try another surgical procedure, the split-lip advancement technique as described by Mutaf. The authors report here on 8 operated children out of 25 patients followed up for Van der Woude syndrome. They evaluated the results according to the appearance of the lip when relaxed, its functional aspect, and the satisfaction of the parents or the child. Our results point to the importance of early surgical treatment, radical excision of dysplastic tissue and the repair of lip muscles to restore good lip functionality and satisfactory bilabial symmetry. The technique now used in our department has made it possible for us to achieve these goals. PMID- 12776240 TI - Lymphangioma colli--a new classification contributing to prognosis. AB - The aim of the study was to introduce a classification of lymphangioma colli, which allows us to predict the expected morbidity and prognosis with surgical treatment. We defined 4 types, dependent on the volume of the lymphangioma as assessed by prenatal ultrasound or clinically: type I included tumors which had no or only a minimal effect on the contour of the neck; type II lymphangiomas were smaller than a line drawn at the lateral border of the head; type III tumors exceeded this line; in type IV the lymphangioma extended beyond the midline of the body. We report on the pre- and postoperative findings in 36 cases operated on between 1988 and 2000, mean follow-up was 4.2 years. 25 % of the cases were classified as type I, 36 % as type II, 22 % as type III, and 17 % as type IV. Cesarean section was carried out for 79 % of type III and IV lymphangiomas following prenatal diagnosis (71 %) and for 14 % of type I and II. MRT was indicated in 38 % of type II compared to 86 % in type III and IV lymphangiomas. Endoscopy was necessary in 25 % of type III and in 66 % of type IV lymphangiomas. In 2 cases (type I and IV), imaging revealed an intrathoracic portion. In 64 % of type I and II cases, resection was complete compared to 21 % in infants with more extended tumors. In all type IV cases, at least one further operation was required, likewise in 25 % of type III lymphangiomas. Postoperative effusion and infection occurred significantly more often in group IV than in groups I to III. 5 nerval lesions remained in children with operated type IV lymphangioma and 2 in type III cases. A visible lack of facial symmetry was evident at the follow-up examination in 83 % of type IV cases and moderate asymmetry was found in 19 % of all type I and II cases. Tracheostomy and gastrostomy were required only in type IV. The introduced classification of lymphangioma colli enables predictive statements to be made concerning the morbidity and prognosis with surgical therapy. The prognosis in type I is excellent; in type II, a cosmetically significant facial asymmetry remains in only 15 %. In type III lymphangiomas postoperative complications occurred in 38 % of cases, the necessity for reoperation with definitive lesions occurred in only 25 %. Morbidity gets significantly higher and the prognosis is worse if the lymphangioma exceeds the midline of the body. PMID- 12776241 TI - Guidelines for the successful treatment of lymphangioma with OK-432. AB - BACKGROUND/PURPOSE: Cystic hygroma or lymphangioma (LA) is a disfiguring benign lesion commonly seen in the neck and face regions in children. Extensive neck/face resectional surgery is frequently performed for this condition, often with disappointing results. An attractive alternative to surgery is injection, sclerosing therapy. OK-432 injection therapy has been characterised as a novel treatment of LA since 1987. Since this first report, there have been a number of articles from different institutions reporting variable success rates with its use. This has resulted in uncertainty and confusion among physicians, surgeons and parents alike as to in whom, when, and how to treat patients with this modality. METHOD: A prospective study over a five-year period. Thirty-five patients were injected with this agent, 1 to 4 times, depending on response. RESULT: In patients with macrocystic LA, complete regression was achieved in 96 %. Patients with microcystic LA responded poorly or not at all. CONCLUSION: The anatomical appearance of the malformation on computerised tomography (or sonar) scan is crucial in determining the treatment strategy taken; macrocystic lesions respond almost universally to OK-432 injections, whereas patients with microcystic disease generally do not respond and should therefore not be injected with this agent. PMID- 12776242 TI - Congenital lobar emphysema: a clinicopathologic evaluation of 14 cases. AB - Controversy still exists concerning the diagnosis and treatment of congenital lobar emphysema (CLE). Although surgical removal of the affected lobe is the most commonly accepted form of treatment, detection of milder or even asymptomatic cases is usually followed by a more conservative management of patients, i. e. non-surgical treatment and follow-up. We therefore decided to evaluate our patients with CLE, placing special emphasis on treatment and diagnostic techniques. We also evaluated quantitative analyses of alveolar diameters. Fourteen children with CLE were analysed retrospectively, including age, sex, clinical picture, localisation, diagnostic and surgical modalities and histopathologic diagnosis. The alveolar diameters of affected lobes were compared with those of the lobectomised patients with other non-obstructive respiratory diseases. All children but one had severe respiratory distress as an initial symptom. All patients, except newborns, had a history of pulmonary infection. All cases underwent thoracic CT examination as the main radiologic method. In all of the patients, only one lobe was affected. We found an obvious mediastinal shift and atelectasis of adjacent lobes due to compression of the affected lobe. The affected lobe was therefore surgically removed in all of the children. In one case, we had to carry out a partial lobectomy to reduce the duration of the operation, due to an intraoperative fall of oxygen saturation. All of the children had an uneventful clinical course postoperatively. The alveolar diameters of the cases with CLE were significantly greater than those of the control patients. We think that the majority of cases with CLE have too severe respiratory distress to avoid surgical removal of affected lobe. Conservative management should be reserved only for patients with milder symptoms or no distress at all. PMID- 12776243 TI - Retroperitoneoscopic approach to the adrenal glands and retroperitoneal tumours in children: where do we stand? AB - Choosing the best way to approach adrenal gland and retroperitoneal tumours is still difficult. We reviewed our first 10 cases operated on by retroperitoneoscopy and compared this approach with other possible ways described in the literature. There were 2 intraoperative complications: 1 opening of the diaphragm and 1 bleeding. Tumour resection was always complete. There was no conversion. There were no postoperative complications. The retroperitoneoscopic approach for adrenalectomy and retroperitoneal tumour resection is increasingly being used. In children, operation is quite fast, without much blood loss and with spectacular postoperative recovery results. Even for the right side we advocate this approach, due to the particular anatomy (small tumour size, less fat, thinner muscle layers) in this age group. Trained surgeons are, of course, mandatory. PMID- 12776244 TI - Abdominal tuberculosis in children. AB - Abdominal tuberculosis (AT) is a rare cause of intraabdominal infection. Surgical intervention is rarely indicated, other than obtaining a specimen for histopathological diagnosis or for the treatment of complications. METHODS: The medical records of 14 patients who were operated on after the diagnosis of AT between 1983 and 2000 were reviewed retrospectively. RESULTS: Median age was 7 years (6 months to 10 years). The presenting clinical signs and symptoms were as follows: colicky abdominal pain (9), weight loss (8), abdominal mass (6), vomiting (5), and night fever (5). Two patients were operated at another centre and referred to our department with faecal fistula and severe malnutrition. Twelve patients were operated on, while diagnostic laparoscopy was performed in two. In uncomplicated cases, surgical intervention was limited to sampling of peritoneal tissue, lymph node and ascites. The reasons for surgical intervention were intestinal obstruction (9), abdominal mass and ascites (6), psoas abscess (1) and intussusception (1). Adhesive peritonitis and ileal loops were the cause of abdominal mass. Necrosis of the bowel (2) and perforation (1) were detected in three patients. The diagnosis was confirmed either by histopathological or microbiological examination. In eight patients, AT was defined at the intestinal mesentery, in three patients it was localised to the peritoneum and in two patients the disease was diffuse. All patients except one with faecal fistula survived and were treated successfully with antituberculous therapy (isoniasid, rifampicin, streptomycin and pyrazinamide combination). CONCLUSION: The diagnosis of AT is difficult before presentation with complications of intraabdominal infection. Since the response to chemotherapy is usually excellent in patients with suspected AT, aggressive surgery should be avoided and initial surgical intervention should be limited to tissue and/or fluid sampling. PMID- 12776245 TI - The contribution of modern imaging to planning separation strategies in conjoined twins. AB - BACKGROUND/AIM: An accurate knowledge of the often complex and unexpected anatomy of conjoined twins is required in order to devise appropriate surgical strategies for separation. The recent progress of imaging techniques prompts an assessment of their contribution to planning separation strategies. MATERIAL AND METHODS: From 1990, we treated 8 consecutive sets of conjoined twins: 1 acardiac acephalus, 1 epigastric heteropagus, 2 omphalopagus, 2 omphalothoracopagus, 1 ischiopagus tetrapus and 1 parapagus tripus. The first two sets were separated straightforwardly after birth and were excluded. Ultrasonographic and plain and contrast X-ray studies were used in all the remaining sets, angiography in 5, CT in 4, and MRI in 4. Helical CT and magnetic resonance angiography (MRA) were used in 3 sets. RESULTS: Two sets of omphalopagus twins and one of omphalothoracopagus twins were separated after birth due to brain damage of one twin, cloacal exstrophy and large arterial shunting, respectively. They were assessed using barium studies (3 sets), IVP (3), angiography (2), CT (2) and MRI (1). Three of six babies survived after separation. In the second set of omphalothoracopagus ecocardiography and MRA revealed that the extent of cardiovascular sharing precluded separation. In the remaining two sets of twins respectively ischiopagus and parapagus, the extent of organ sharing was depicted using three-dimensional helical CT and MRI; the four babies are alive after separation. CONCLUSIONS: Conventional imaging contributes only modestly to separation planning. In contrast, advanced imaging techniques are amazingly efficient in depicting complex fusions that have to be accurately recognised when planning realistic separation strategies. PMID- 12776246 TI - Treatment dilemma: conservative versus surgery in cutis aplasia congenita. AB - Cutis aplasia congenita (CAC), a congenital absence of skin and its appendages, may extend into underlying muscles and bones. The scalp is the commonest site and it may be associated with acrania. CAC presents either as a thin transparent membrane, a black eschar, an ulcer or a healed scar. The dilemma of either immediate surgical management or conservative treatment is much more pronounced in the presence of acrania. Two patients with scalp lesions measuring 12 x 8 cm and 14 x 12 cm respectively and one patient with 4 cm wide circumferential trunk cutis aplasia treated conservatively are presented. The conservative treatment is simple, easy to carry out, and effective even for large defects; therefore, it is recommended in cutis aplasia congenita till complete healing. Surgical interventions such as tissue expansion and resurfacing, contracture release, etc. are for the correction of subsequent deformity at a later date. PMID- 12776247 TI - Antenatal detection of a communicating duodenal duplication. AB - We report a case of a subhepatic cystic mass diagnosed in utero by antenatal ultrasonography (US) at 15 weeks' gestation which subsequently proved to be a communicating duodenal duplication. In this male foetus, the differential diagnosis was choledochal cyst, congenital biliary atresia, foregut duplication or omentum cyst. Neonatal US examination lead to a diagnosis of duodenal duplication, also confirmed by barium gastrointestinal series. He was operated on day 8 and recovered uneventfully. We discuss the accuracy of antenatal US in the diagnosis of such right upper quadrant cystic masses. Now that antenatal findings are becoming increasingly sensitive in the detection of foetal anomalies, and parents need to be informed about the suspected pathology and its prognosis, we tried to determine, in the light of this case and a review of the literature, how antenatal US findings can offer more accuracy in the diagnosis of duodenal duplication. PMID- 12776248 TI - Congenital fibrosarcoma of the jejunum in a premature infant with meconium peritonitis. AB - We report an unusual case of perforation of a congenital fibrosarcoma of the jejunum in utero and secondary meconium peritonitis. Prenatal ultrasound showed polyhydramnios and fetal ascites from 25 gestational weeks in the absence of other fetal congenital anomalies. A 2200 g baby girl was born at 34 weeks gestation, presenting with severe generalized edema and respiratory distress immediately after birth. Plain radiography revealed progressive abdominal distension and pneumoperitoneum. The baby subsequently underwent surgery at the age of one day. A perforation of the upper jejunum, which had resulted in meconium peritonitis, was discovered intraoperatively and the perforated section of the intestine was resected and anastomosed successfully. The postoperative course was uneventful. Pathological examination confirmed that the perforation was caused by rupture of a congenital fibrosarcoma originating from the jejunum. Rupture of a malignant tumor is an extremely rare cause of peritonitis in the fetus and neonate. PMID- 12776249 TI - Argon plasma coagulation for blue rubber bleb nevus syndrome in a female infant. AB - We report on the case of an 8-month-old baby with blue rubber bleb nevus syndrome (BRBNS) presenting with recurrent bleeding from multiple haemangiomas in the colon and rectum. Treatment with Argon plasma coagulation resulted in long-term success. Our case is unique in that 1) it is the first documented case treated by argon plasma coagulation; 2) it is only the second case in the world literature with involvement of the genital tract; 3) it is the youngest patient treated endoscopically to date; 4) it is the endoscopically treated case with the longest follow-up; 5) it represents the second case of BRBNS reported from this part of the world; and 6) it expands the spectrum of mucosal lesions amenable to this new form of treatment. Argon plasma coagulation is a simple, inexpensive and effective treatment for selected lesions in BRBNS. PMID- 12776250 TI - Volvulus of the transverse colon in a child: a case report. AB - A case of transverse colon volvulus in a 10-year-old female child with trisomy 13 is reported, bringing the total number of cases of children reported in the English language medical literature to 14. Although this type of volvulus is rare, a definite pattern is noticeable, so we should suspect its diagnosis in a patient with chronic constipation, mental retardation and motor disturbances. Resection of the involved segment and primary anastomosis is the definite treatment. PMID- 12776251 TI - IBD5 is a general risk factor for inflammatory bowel disease: replication of association with Crohn disease and identification of a novel association with ulcerative colitis. AB - Inflammatory bowel disease (IBD) refers to complex chronic relapsing autoimmune disorders of the gastrointestinal tract that have been traditionally classified into Crohn disease (CD) and ulcerative colitis (UC). We have previously reported that genetic variation within a 250-kb haplotype (IBD5) in the 5q31 cytokine gene cluster confers susceptibility to CD in a Canadian population. In the current study, we first replicated this association by examining 368 German trios with CD and demonstrating, by transmission/disequilibrium testing (TDT), that the same haplotype is associated with CD (chi2=5.97; P=.007). Our original association study focused on the role of IBD5 in CD; we next explored the potential contribution of this locus to UC susceptibility in 187 German trios. Given the TDT results in the present cohort with UC, IBD5 may also act as a susceptibility locus for UC (chi2=8.10; P=.002). We then examined locus-locus interactions between IBD5 and CARD15, a locus reported elsewhere to confer risk exclusively to CD. Our current results indicate that the two loci act independently to confer risk to CD but that these two loci may behave in an epistatic fashion to promote the development of UC. Moreover, IBD5 was not associated with particular clinical manifestations upon phenotypic stratification in the current cohort with CD. Taken together, our results suggest that IBD5 may act as a general risk factor for IBD, with loci such as CARD15 modifying the clinical characteristics of disease. PMID- 12776253 TI - Gallium nitrate revisited. AB - Gallium nitrate, the nitrate salt of the "near-metal" element gallium, is highly effective in the treatment of cancer-related hypercalcemia. Unlike bisphosphonates, gallium nitrate is effective in both parathyroid hormone-related protein-mediated and non-parathyroid hormone-related protein-mediated hypercalcemia. Gallium nitrate's effects on bone are clearly different from those of bisphosphonates. Gallium nitrate enhances calcium and phosphate content of bone and has direct, noncytotoxic effects on osteoclasts at markedly lower doses than those used for the treatment of cancer-related hypercalcemia. The drug may have clinical application in a variety of disorders associated with accelerated bone loss, including multiple myeloma. Gallium nitrate was originally evaluated as an antitumor agent. Its antitumor activity occurs at somewhat higher doses than those used in the treatment of cancer-related hypercalcemia. Gallium nitrate has substantial single-agent activity in the treatment of advanced lymphoma, particularly diffuse large cell lymphoma, small lymphocytic lymphoma, and follicular lymphoma. Because of its profile, including a different mechanism of action and minimal myelosuppression, the drug merits further evaluation in the treatment of advanced lymphoma. Gallium nitrate also has activity in advanced bladder cancer and may be useful in patients with metastatic or unresectable disease failing first-line chemotherapy regimens. Gallium nitrate exhibits a range of dose-dependent pharmacologic actions that provide a basis for its therapeutic potential in a variety of diseases and warrants further investigational evaluation as an antiresorptive and antitumor agent. PMID- 12776252 TI - Ehlers-Danlos syndrome with severe early-onset periodontal disease (EDS-VIII) is a distinct, heterogeneous disorder with one predisposition gene at chromosome 12p13. AB - Ehlers-Danlos VIII (EDS-VIII) is an autosomal dominant disorder characterized by severe early-onset periodontal disease in conjunction with the features of Ehlers Danlos syndrome (EDS). We performed a genomewide linkage search in a large Swedish pedigree with EDS-VIII and established linkage to a 7-cM interval on chromosome 12p13, generating a maximum multipoint LOD score of 5.17. Analysis of four further pedigrees with EDS-VIII revealed two consistent with linkage to 12p13 and two in which linkage could be excluded, indicating that EDS-VIII is a genetically heterogeneous disorder. Chromosome 12p13 has not previously been implicated in either EDS or periodontal disease and contains no known collagen genes or collagen-processing enzymes. Mutational screening of the microfibril associated glycoprotein-2 gene, a strong candidate within the minimal interval, did not reveal any likely pathogenic mutations. PMID- 12776254 TI - The effects of gallium nitrate on bone resorption. AB - Gallium nitrate has been shown to be an effective treatment for patients with cancer-related hypercalcemia. Clinical studies have also suggested the drug may have considerably broader use in other diseases associated with accelerated bone loss including multiple myeloma, bone metastases, Paget's disease, and osteoporosis. The actions of gallium nitrate on bone are quite distinct from those of bisphosphonates. Preclinical studies show that gallium preferentially accumulates in trace amounts in metabolically active regions of bone. When present, gallium favorably alters the mineral properties to enhance hydroxyapatite crystallization and reduce mineral solubility. The drug also acts on the cellular components of bone to reduce bone resorption by decreasing acid secretion by osteoclasts. This effect appears to be mediated by inhibition of the ATPase-dependent proton pump of the osteoclast's ruffled membrane. Gallium does not inhibit the development or recruitment of osteoclasts to bone tissue, unlike many bisphosphonates that may induce osteoclast apoptosis. Together, these pharmacologic actions may yield a skeletal system with increased calcium and phosphate content and improved biomechanical strength. Gallium nitrate has potent antiresorptive effects on bone that can be achieved at considerably lower doses than are currently used for cancer-related hypercalcemia. Parenteral and oral formulations of gallium appear to have high activity in bone resorptive disorders, and thus development should be vigorously pursued in these diseases. PMID- 12776255 TI - Treatment of cancer-related hypercalcemia: the role of gallium nitrate. AB - Hypercalcemia is a common, life-threatening metabolic disorder that can be associated with cancer. Its pathophysiology includes enhanced osteoclastic bone resorption and decreased renal excretion of extracellular calcium. Symptoms of hypercalcemia include nausea, vomiting, bone pain, polyuria, renal insufficiency, bradycardia, and arrhythmia. The goals of medical therapy are to inhibit bone resorption and promote renal calcium excretion. Hydration is the first step in management. Treatments for hypercalcemia include phosphates, calcitonin, bisphosphonates, and gallium nitrate. Although intravenous phosphates prevent intestinal calcium absorption and inhibit mineral and bone matrix resorption, serious adverse events include renal failure, hypotension, extraskeletal calcification, and severe hypocalcemia. Calcitonin has a rapid onset of action and can lower serum calcium concentrations within hours, but its usefulness is limited by its short duration of effect and the development of tachyphylaxis. Bisphosphonates are effective inhibitors of bone resorption but appear to have decreased response rates in hypercalcemic patients with high levels of parathyroid-related protein. Gallium nitrate, an antitumor agent noncytotoxic to osteoclasts and bone cells, appears to be more effective than pamidronate, etidronate, and calcitonin in the treatment of cancer-related hypercalcemia. Importantly, unlike bisphosphonates, gallium nitrate is effective in both parathyroid-related protein-mediated and non-parathyroid-related protein-mediated hypercalcemia. PMID- 12776256 TI - Gallium nitrate in multiple myeloma: prolonged survival in a cohort of patients with advanced-stage disease. AB - Multiple myeloma is characterized by bone destruction mediated by osteoclastic bone resorption. Skeletal complications of myeloma, including bone pain, fractures, spinal cord compression and hypercalcemia, result in significant morbidity. Gallium nitrate was shown in a small, randomized trial to attenuate the rate of bone loss in patients with myeloma treated with chemotherapy. In a retrospective analysis, we found that patients with advanced multiple myeloma treated with chemotherapy plus gallium nitrate had markedly prolonged median survival compared with similar patients treated with chemotherapy alone (87+ months v 48 months, respectively). These data suggest that gallium nitrate may have a positive, indirect benefit on survival in myeloma by decreasing the rate of bone resorption. Further evaluation of gallium nitrate to attenuate progression of disease in patients with multiple myeloma is warranted. PMID- 12776257 TI - Gallium nitrate in the treatment of lymphoma. AB - Gallium nitrate is effective and well tolerated for the treatment of cancer related hypercalcemia. At somewhat higher doses, gallium nitrate also has cytotoxic activity against a variety of cancers. The probable mechanism is inhibition of both ribonucleotide reductase and a protein tyrosine phosphatase. Radioactive gallium ((67)Ga) is concentrated at sites of malignant lymphoma, Hodgkin's disease, and other tumors. Gallium nitrate has substantial single-agent activity in the treatment of patients with advanced lymphoma and has also shown activity when used in combination with other agents. Significant response rates have been observed in patients with diffuse large cell lymphoma, small lymphocytic lymphoma, and follicular lymphoma. Because of its unique mechanism of action, gallium nitrate could be non-cross-resistant with many of the cytotoxic agents used as standard chemotherapy for non-Hodgkin's lymphoma. Nephrotoxicity, the most frequent adverse event associated with gallium nitrate, can generally be minimized by ensuring adequate oral hydration and avoiding concomitant use of other nephrotoxic drugs. Gallium nitrate causes little myelosuppression and is therefore well tolerated by patients with advanced disease who have received extensive prior therapy. Given its unique mechanism of action, the high level of single-agent activity in published clinical trials, the absence of significant myelosuppression, and the potential lack of cross-resistance, further clinical study of gallium nitrate both alone and in combination with other active agents is warranted. PMID- 12776258 TI - Gallium nitrate in the treatment of bladder cancer. AB - For over 15 years, the MVAC regimen (methotrexate/vinblastine/doxorubicin/cisplatin) has been standard chemotherapy for patients with unresectable or metastatic bladder cancer. The taxanes and gemcitabine have provided new treatment options, but development of more effective regimens is needed. Gallium nitrate has significant activity as a single agent in the treatment of advanced bladder cancer, including activity in heavily pretreated patients and patients previously treated with MVAC or single agent cisplatin. At a dosage of 300 mg/m(2) daily by continuous infusion for 5 to 7 days every 3 weeks, toxicity has been acceptable in the treatment of patients with refractory disease. Gallium nitrate is also active in combination regimens for advanced bladder cancer. Because it has a different mechanism of action, minimal myelosuppression, and activity in previously treated patients, gallium nitrate may be useful as a single agent in patients with advanced bladder cancer who fail front-line chemotherapy regimens. Evaluation of gallium nitrate in combination with newer agents such as the taxanes or gemcitabine may also be warranted given its activity, different mechanism of action, and non-overlapping toxicity profile. PMID- 12776259 TI - A substantive theory of Keeping the Spirit Alive: the Spirit Within children with cancer and their families. AB - Despite increasing survivorship, childhood cancer is nonetheless still a very traumatic phenomenon. Children and families must confront many new stressors and challenges. Understanding what families experience is essential if pediatric oncology nurses hope to provide sensitive and comprehensive care. To help discover meaningful descriptions and interpretations of families' experiences with childhood cancer, process-oriented qualitative research needs to be undertaken. A longitudinal qualitative study guided by the philosophy of interpretive interactionism was conducted to arrive at an understanding of childhood cancer from the children's and families' perspectives. A theoretical purposive sample of 39 families residing in western Canada participated. Children (4(1/2)-to-18 years of age) with a variety of cancer diagnoses and their families (parents and siblings) participated. Data collection methods included formal and informal interviewing and participant observation. Data were analyzed by the constant comparative method of grounded theory and analysis of illness narratives. Although the original intent of this study was to focus on the children's cancer symptoms, the children and families could not easily separate their cancer symptom experiences from the whole cancer experience and all its associated "rough spots." They first needed to talk about the overall cancer experience, which led to the development of the study's first substantive theory- Keeping the Spirit Alive: The Spirit Within. This substantive theory provided an understanding of how children and families used the process of keeping the spirit alive in response to them having to experience the phenomenon of the cancer: "getting through all the rough spots." Description of this theory is the focus of this article. Ways that nurses can assist children and families in maintaining a sense of spirit are presented. PMID- 12776260 TI - Distressing events for children and adolescents with cancer: child, parent, and nurse perceptions. AB - Distressing events for children with cancer (N = 121), 0 to 19 years of age, were investigated. Data were gathered through interviews with 50 children, 65 parents, and 118 nurses. Each participant was asked: "Has there been any especially distressing event for you/your child/the child with regard to disease and treatment?" Data were analyzed by content analysis. The categories that emerged from the analysis were grouped into a physical and an emotional dimension. The most frequently mentioned aspects of distress referred to the physical dimension: pain resulting from diagnostic procedures and treatments, nausea, and fatigue. The most frequently mentioned physical aspect of distress was, for children 0 to 3, 4 to 7, and 8 to 12 years of age, pain resulting from diagnostic procedures and treatments, and for children > or =13 years of age, nausea. The most frequently mentioned aspects of distress referred to the emotional dimension were categorized as confinement, feeling of alienation, and worry before medical procedures. The most frequently mentioned emotional aspect of distress was, for children 0 to 3 years of age, confinement; 4 to 7 years of age, feeling of alienation; 8 to 12 years of age, worry about death; and > or =13 years of age, changed appearance. For children 0 to 3, 4 to 7, and > or =13 years of age, aspects of distress of a physical character were mentioned most frequently. For children 8 to 12 years of age, aspects of distress of an emotional character were mentioned most frequently. PMID- 12776261 TI - Childhood brain tumors: children's and siblings' concerns regarding the diagnosis and phase of illness. AB - Our objective was to identify commonly reported problems and helpful resources important to children with brain or spinal cord tumors and siblings during phases of illness. Affected children and siblings from 40 families responded to a regional survey. Items encompassed four general content areas: health care provider interactions, medical information/education, health care utilization, and psychosocial. Children rated individual items as helpful, a problem, and for importance; relative frequencies were derived. One-third or more affected children reported important problems: hospitalization/surgery--help with schoolwork; hospital discharge--help with changes in physical activity, appearance, moods, special needs at school; adjuvant treatment--Internet information, help with moods; and remission--socializing. One-third or more siblings reported important problems: diagnosis--information about etiology and prognosis, manner of physician and parents in providing information; hospitalization/surgery--information about prognosis, help with schoolwork; hospital discharge--help with schoolwork; adjuvant treatment--help with changes in sister or brother's appearance, physical activity, moods, and information about the treatment; end of life--treatment of sister's or brother's pain, information about dying, family harmony, support from friends, help with schoolwork, and preparation for the death. Consideration of problems and helpful resources will provide the framework for developing and evaluating intervention strategies at each phase of illness. PMID- 12776262 TI - Research column: protection of children as subjects in research. PMID- 12776263 TI - Alona's farewell. PMID- 12776264 TI - An "evidence-based" survey of therapeutic options for IgA nephropathy: assessment and criticism. AB - BACKGROUND: Immunoglobulin A (IgA) nephropathy is a worldwide disease that causes end-stage renal failure in 15% to 20% of patients within 10 years of the apparent onset of disease and in 30% to 40% of individuals within 20 years. Severity of renal lesions, serum creatinine level, and severe proteinuria are adverse prognostic indicators. No specific treatment has been established, but several approaches have been experimented. METHODS: We reviewed the literature and evaluated the quality of published randomized trials using standard methods and the quality of their reporting according to the revised version of the Consolidated Standards for Reporting Trials Statement. Meta-analyses of randomized trials on the efficacy of steroid treatment, cytotoxic agents, and fish oils on the outcome of renal function and daily proteinuria in patients with IgA nephropathy were performed. RESULTS: Only 10 randomized trials were available and included in the review. Their quality was very poor, and a limited amount of data was reported. Cytotoxic agents seem beneficial on both renal function (relative risk, 0.38; 95% confidence interval [CI], 0.22 to 0.66) and daily proteinuria (weighted mean difference [WMD], -1.16; 95% CI, -2.18 to -0.14) in patients with moderate to severe renal damage, steroids act mainly on proteinuria (WMD, -0.50; 95% CI, -0.78 to -0.21), and fish oils do not imply a particular benefit. This statement is based on the very limited and poor available published evidence. CONCLUSION: Only a few randomized trials, of low quality and inadequately reported, are available relating to treatment of IgA nephropathy. More properly designed and reported trials are necessary to reach a definitive assessment of this matter. PMID- 12776265 TI - Significance of endothelial cell survival programs for renal transplantation. AB - Initial and longer term kidney transplant function is determined in part by the renal allograft microcirculation because it provides a thromboresistant surface, regulates cellular infiltration, and elaborates paracrine and autocrine growth and survival factors. Loss of endothelial-derived signaling mediators accelerates vascular injury and endothelial cell (EC) death. EC apoptosis is implicated in accelerated allograft vasculopathy and premature loss of organ function. Renal allograft EC injury and replacement by recipient-derived repair mechanisms has long been proposed to influence allograft acceptance and function. Repair of cellular injury in allografts is linked with cell-survival mechanisms, but few precise indicators exist to predict recovery and repair in organ transplants. The significance of the growth phenotype of the microvascular endothelium for acute and longer term renal allograft survival is presented. PMID- 12776266 TI - A randomized study of two long-course prednisolone regimens for nephrotic syndrome in children. AB - BACKGROUND: Long-course prednisolone regimens have been shown to be more effective than short-course regimens in sustaining remission of nephrotic syndrome in children. However, the most beneficial approach among the long-course regimens remains unknown. METHODS: Seventy-three children with new-onset nephrotic syndrome were allocated at random to the two long-course regimens and followed up for 2 years. Group A was administered prednisolone at a daily dose of 60 mg/m2 for 6 weeks, followed by an alternate-day dose of 40 mg/m2 for 6 weeks (the long daily regimen). Group B was administered the same daily dose for 4 weeks, followed by an alternate-day dose of 60 mg/m2 for 4 weeks, and doses were tapered by 10 mg/m2 every 4 weeks (the long alternate-day regimen). RESULTS: Group B had a lower incidence of corticosteroid toxicities than group A during the initial treatment. Kaplan-Meier analysis of the sustained remission rate of the two treatment groups showed a marginally significant difference (P = 0.069) and showed a significant difference when patients were stratified for age of disease onset (P = 0.048). In a subgroup of younger children (<4 years at onset), group B had a greater rate of sustained remission (P < 0.01) and fewer children with frequent relapses (P < 0.05) than group A, whereas in older children (> or =4 years at onset), both groups had similar good sustained remission rates. CONCLUSION: These findings collectively indicate that the long alternate-day regimen may be more beneficial, with less corticosteroid toxicities, than the long daily regimen, and children with younger age at disease onset may be susceptible to relapse and especially benefit from the long alternate-day regimen for sustaining remission of the disease. PMID- 12776267 TI - Are children with idiopathic nephrotic syndrome at risk for metabolic bone disease? AB - BACKGROUND: Children with idiopathic nephrotic syndrome (INS) may be at risk for metabolic bone disease (MBD) because of biochemical derangements caused by the renal disease, as well as steroid therapy. No large study to date has shown conclusively that these children are prone to MBD. METHODS: We prospectively studied 100 consecutive children with INS for clinical, biochemical, and radiological evidence of MBD. These children were treated with prednisone as follows: initial episode, prednisone, 60 mg/m2/d for 6 weeks, followed by 40 mg/m2 on alternate days for 6 weeks. Relapses were treated with 60 mg/m2/d until remission for 3 days, followed by 40 mg on alternate days for 4 weeks and tapered by 10 mg/m2/wk. Osteoporosis is defined as a bone mineral density (BMD) value evaluated by dual-energy X-linked absorptiometry of the lumbar spine of a z score of 2.5 SDs less than the mean. Univariate and multivariate analyses were performed to analyze for factors predictive of low BMD z score. Children were divided into two groups: those who had received repeated courses of steroid therapy (group II: frequent relapsers (FRs), steroid dependent (SD), or steroid nonresponders (SNRs) versus those who had received infrequent courses (group I: infrequent relapsers). RESULTS: Twenty-two of 100 children (22%) had osteoporosis. Comparing clinical features, we observed that 6 of 70 children in group II were symptomatic (hypocalcemic signs) compared with none of 30 children in group I (P = 0.10). However, children in group II had significantly lower mean BMD z scores compared with group I (-1.65 +/- 1.35 versus -1.08 +/- 1.0; P = 0.01). Also, 20 of 70 children in group II had osteoporosis compared with 2 of 30 children in group I (P = 0.012). Children in group II had been administered significantly greater doses of steroids compared with group I (P < 0.00001). On multivariate analysis, factors predictive of a low BMD score were older age at onset (P = 0.000), lower total calcium intake (P = 0.000), and greater cumulative steroid dose (P = 0.005). CONCLUSION: Children with INS are at risk for low bone mass, especially those administered higher doses of steroids (FRs, SD, or SNRs). These children should undergo regular BMD evaluations, and appropriate therapeutic interventions should be planned. PMID- 12776268 TI - Clinical, histopathologic, and genetic studies in nine families with focal segmental glomerulosclerosis. AB - BACKGROUND: Familial forms of focal segmental glomerulosclerosis (FSGS) are caused by mutations in genes at 1q25-31 (gene for steroid-resistant nephrotic syndrome 2 [NPHS2]), 11q21-22, 19q13 (gene for alpha-actinin 4 and NPHS1), and at additional unidentified chromosomal loci. METHODS: We describe clinical and histopathologic features and results of linkage analysis in nine consecutive index cases with familial FSGS who, together with their families, were referred for genetic studies. RESULTS: Two of the index cases presented in childhood (22%) and seven cases presented in adolescence or adulthood (78%). Six of their families (67%), including the two cases with childhood-onset disease, showed probable autosomal recessive inheritance. FSGS segregated at the 1q25-31 locus in two of these families and at the 11q21-22 locus in four families. None had disease caused by mutations in genes at the 19q13 locus, and no locus was identified in the three remaining families. Clinical features of proteinuria, minimal hematuria, hypertension, preeclampsia, and progressive renal impairment were usually present with autosomal recessive or dominant inheritance and with disease that segregated at the different loci. Eighteen renal biopsies from affected members of eight families showed a strong correlation between tubulointerstitial damage and percentage of obsolescent glomeruli (rho = +0.76; P < 0.01). None of the 13 patients from eight families who underwent transplantation developed recurrent FSGS in their grafts. In general, carriers of autosomal recessive disease had no distinctive clinical features apart from the development of preeclampsia in successive pregnancies. CONCLUSION: Familial forms of FSGS are not uncommon, and presentation frequently is in adolescence or adulthood, even when inheritance is autosomal recessive. Furthermore, carriers of autosomal recessive FSGS often have no distinctive phenotype. PMID- 12776269 TI - Glomerular basement membrane length to podocyte ratio in human nephronopenia: implications for focal segmental glomerulosclerosis. AB - BACKGROUND: This study aims to probe a possible mechanism for the development of focal segmental glomerulosclerosis (FSGS) in human nephronopenic states. METHODS: To this end, length (in microns) of the glomerular basement membrane (GBM) covered by the cytoplasm of a podocyte was determined in controls and various congenital and acquired forms of nephronopenia with and without FSGS. RESULTS: Results indicate that for all nephronopenic groups without FSGS, the respective lengths are not significantly different from those of controls. With the exception of long-surviving renal allografts with FSGS, each nephronopenic group with FSGS has a significantly longer length compared with its respective control group without FSGS. Although the corresponding length in long-surviving allografts with FSGS is longer than that in the transplant group without FSGS, the difference does not attain significance. Among all groups with FSGS, the respective GBM lengths are not significantly different among cases of congenital causes of nephronopenia, but are significantly longer than when nephronopenia is acquired, whereas the corresponding lengths among uninephrectomized and transplant groups are not significantly different from one another. CONCLUSION: These results support the hypothesis that in enlarging glomeruli of nephronopenia, the dominant reaction of podocytes is to undergo hypertrophy. The capacity for this is greater in podocytes of native kidneys than allografts, being most pronounced when the nephronopenia is congenital than if acquired, and being most constrained in allografts. This then imposes a limit to glomerular enlargement in nephronopenia beyond which denudation of segments of the enlarging/elongating glomerular capillaries occurs, when FSGS supervenes. PMID- 12776270 TI - Methylenetetrahydrofolate reductase gene polymorphism and susceptibility to diabetic nephropathy in type 1 diabetes. AB - BACKGROUND: The T allele of the C677T polymorphism in the methylenetetrahydrofolate reductase (MTHFR) gene is associated with elevated plasma homocysteine levels, and it has been postulated to be a risk factor for the development of diabetic nephropathy. We examined this hypothesis in both a case-control and a follow-up study in individuals with type 1 diabetes. METHODS: In the case-control study, the control group included 310 subjects with normoalbuminuria and diabetes duration of 15 years or greater, and the case group included 88 prevalent cases with end-stage renal disease (ESRD). The follow-up study included 235 subjects with overt proteinuria followed up for 6 years (on average), during which time ESRD developed in 69 subjects. DNA from each individual was genotyped for the C677T MTHFR polymorphism. RESULTS: The frequency of TT homozygotes did not vary significantly among the four groups: 10% in controls, 15% in prevalent cases of ESRD, 13% in cases with new-onset ESRD, and 11% in those who remained proteinuric during follow-up (P = 0.9, 6 df). Similarly, frequency of the T allele varied little among the same groups (range, 33% to 36%; P = 0.9, 3 df) During follow-up, 52 of 323 individuals with diabetic nephropathy died. Total mortality rates were 4.3/100 person-years in TT homozygotes, 2.4/100 person-years in CT heterozygotes, and 3.0/100 person-years in CC homozygotes (P = 0.55, 2 df). CONCLUSION: Using both a large case-control and a follow-up study, we found no evidence that the C677T MTHFR polymorphism has a significant role in the development of diabetic nephropathy in type 1 diabetes. PMID- 12776271 TI - Prevalence of hematuria among Zuni Indians with and without diabetes: The Zuni kidney Project. AB - BACKGROUND: There is an epidemic of kidney disease among the Zuni Indians. In contrast to other American Indian tribes, the epidemic among the Zuni Indians is attributable to diabetic and nondiabetic renal disease. METHODS: The Zuni Kidney Project, established to reduce the burden of renal disease, conducted a population-based cross-sectional survey of Zuni Indians aged 5 years or older to precisely estimate the prevalence of hematuria. The survey used neighborhood household clusters as the sampling frame to maximize ascertainment and minimize bias. During the survey, we administered a questionnaire; collected blood and urine samples; and measured blood pressure, height, and weight. RESULTS: Age and sex distributions in our sample (n = 1,469) were similar to those of the eligible Zuni population (n = 9,228). Prevalences of hematuria, defined as dipstick of trace or greater and 50 red blood cells/microL or greater, age- and sex-adjusted to the Zuni population aged 5 years or older, were 33.2% (95% confidence interval [CI], 30.7 to 35.6) and 17.8% (95% CI, 15.8 to 19.8), respectively. Hematuria of trace or greater was more common among females (40.6%; 95% CI, 37.0 to 44.1) than males (25.1%; 95% CI, 21.8 to 28.4). Hematuria of trace or greater was common among Zuni Indians without diabetes (females, 39.7%; 95% CI, 35.7 to 43.8; males, 22.7%; 95% CI, 19.4 to 26.1) and with diabetes (females, 47.5%; 95% CI, 39.8 to 55.2; males, 45.8%; 95% CI, 34.3 to 57.3). Diabetes and alcohol use for greater than 10 years were associated with hematuria among males, but not females. CONCLUSION: The prevalence of hematuria is high among Zuni Indians with and without diabetes. These findings are consistent with the hypothesis that nondiabetic kidney disease is common among Zuni Indians with and without diabetes. PMID- 12776272 TI - The expanding spectrum of renal diseases associated with antiphospholipid syndrome. AB - BACKGROUND: The association of thrombotic events and/or pregnancy complications with circulating antiphospholipid antibodies defines antiphospholipid syndrome (APS). In previous reports, renal involvement in APS consisted mainly of thrombotic vascular complications involving large vessels or intrarenal small sized vessels (APS nephropathy). We report 9 cases of glomerulonephritis associated with APS. These cases are characterized by predominant pathological features distinct from vascular APS nephropathy. METHODS: We reviewed consecutive renal biopsies examined in 2 French university hospitals between 1980 and 2002 and identified renal biopsies performed in patients with primary APS. RESULTS: We identified 29 biopsies performed in patients with APS. Twenty biopsies showed characteristic features of APS nephropathy. In 9 cases, predominant pathological features distinct from vascular APS nephropathy were noted: membranous nephropathy (3 cases), minimal change disease/focal segmental glomerulosclerosis (3 cases), mesangial C3 nephropathy (2 cases), and pauci-immune crescentic glomerulonephritis (1 case). In 7 cases, the presentation of renal symptoms was subacute or chronic. Two patients experienced episodes of acute renal failure. At referral, median creatinine clearance was 50 mL/min (0.83 mL/s) (range, 18 to 117 mL/min [0.30 to 1.95 mL/s]). Proteinuria was noted in all cases (range, 1.5 to 15 g/d), with nephrotic syndrome in 4 cases. Lupus anticoagulant was present in all cases, and anticardiolipin antibodies, in 8 cases. Anti-DNA antibodies repeatedly were negative in all cases. Treatment consisted of antihypertensive therapy (6 cases), anticoagulant drugs (5 cases), steroids (4 cases), and antiplatelet drugs (3 cases). At last follow-up, renal function remained stable in 7 patients. Of 2 patients presenting with acute renal failure, 1 patient recovered normal renal function, whereas the other patient progressed to end-stage renal failure. CONCLUSION: The cases reported here represent a new aspect of the expanding spectrum of renal diseases encountered in association with APS. PMID- 12776273 TI - Associations between circulating inflammatory markers and residual renal function in CRF patients. AB - BACKGROUND: Circulating levels of cytokines and other inflammation markers are markedly elevated in patients with chronic renal failure. This could be caused by increased generation, decreased removal, or both. However, it is not well established to what extent renal function per se contributes to the uremic proinflammatory milieu. The aim of the present study is to analyze the relationship between inflammation and glomerular filtration rate (GFR) in 176 patients (age, 52 +/- 1 years; GFR, 6.5 +/- 0.1 mL/min) close to the initiation of renal replacement therapy. METHODS: Circulating levels of high-sensitivity C reactive protein (hsCRP), tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), hyaluronan, and neopterin were measured after an overnight fast. Patients subsequently were subdivided into two groups according to median GFR (6.5 mL/min). RESULTS: Despite the narrow range of GFR (1.8 to 16.5 mL/min), hsCRP, hyaluronan, and neopterin levels were significantly greater in the subgroup with lower GFRs, and significant negative correlations were noted between GFR and IL-6 (rho = -0.18; P < 0.05), hyaluronan (rho = -0.25; P < 0.001), and neopterin (rho = -0.32; P < 0.0005). In multivariate analysis, although age and GFR were associated with inflammation, cardiovascular disease and diabetes mellitus were not. CONCLUSION: These results show that a low GFR per se is associated with an inflammatory state, suggesting impaired renal elimination of proinflammatory cytokines, increased generation of cytokines in uremia, or an adverse effect of inflammation on renal function. PMID- 12776274 TI - Insulin resistance is related to left ventricular hypertrophy in patients with polycystic kidney disease type 1. AB - BACKGROUND: Left ventricular hypertrophy (LVH) is common in patients with autosomal dominant polycystic kidney disease (ADPKD). Although insulin resistance contributes to cardiac hypertrophy, the relationship between insulin resistance and LVH in patients with ADPKD has not been previously studied. METHODS: We performed M-mode and color Doppler echocardiography on 176 family members (106 patients and 70 healthy relatives) from 16 families with polycystic kidney disease type 1 (PKD1). Left ventricular mass index (LVMI) was calculated using the Penn equation and corrected for body surface area. Fasting insulin and glucose concentrations were measured and insulin resistance was evaluated by means of the homeostasis model assessment. RESULTS: In multivariate regression analysis, insulin resistance was significantly associated with LVMI in healthy relatives (P < 0.01) and patients with PKD1 (P < 0.05) independent of age, weight, systolic blood pressure, and albuminuria. CONCLUSION: Insulin resistance is associated with LVMI in patients with PKD1 independently of other factors known to increase LVMI. PMID- 12776275 TI - Hypouricemia in individuals admitted to an inpatient hospital-based facility. AB - BACKGROUND: Decreased serum uric acid levels resulting from renal urate wasting occasionally are reported in hospitalized patients because of isolated or generalized proximal tubular damage. There are limited recent findings with regard to the incidence and cause of hypouricemia in patients admitted to an internal medicine clinic. The aim of this study is to examine the prevalence of hypouricemia in individuals admitted to our inpatient hospital-based facility and identify underlying causes and pathogenetic mechanisms and any association of hypouricemia and uricosuria with other tubular defects. METHODS: A total of 7,250 serum urate measurements were available on patients' admission. Hypouricemia is defined as a serum urate level less than 2.5 mg/dL (149 micromo/L). In all hypouricemic cases, a detailed clinical and laboratory investigation was performed. RESULTS: Hypouricemia was found in 90 patients (1.24%). In all except one patient, hypouricemia was associated with inappropriate uricosuria (urate fractional excretion [FE] > 10%; range, 10.8% to 94%). There was an inverse correlation between serum uric acid level and its FE (r = -0.73; P < 0.0001). The most common causes of hypouricemia were obstructive jaundice of any cause (n = 18), solid or hematologic neoplasias (n = 17), diabetes mellitus (n = 12), drugs affecting urate homeostasis (n = 10), and intracranial diseases (n = 8). Seventeen patients with hypouricemia showed one or more other manifestations of proximal tubular damage, such as glucosuria, inappropriate phosphaturia leading to hypophosphatemia, and kaliuria resulting in hypokalemia. CONCLUSION: Hypouricemia caused by inappropriate uricosuria is not rare in patients admitted to an internal medicine clinic, is related to underlying diseases, and may be associated with other abnormalities of proximal tubular function. PMID- 12776276 TI - Randomized, crossover study of the effect of vitamin C on EPO response in hemodialysis patients. AB - BACKGROUND: Vitamin C has been reported to be an effective adjuvant agent in the treatment of anemia in iron-overloaded hemodialysis patients. We aim to evaluate its effect on erythropoietin (EPO) response in a prospective, randomized, double blind, crossover study. METHODS: Sixty-three patients were randomly divided into two groups. Group 1 was treated with intravenous vitamin C, 500 mg, three times a week, and group 2, with placebo for 6 months. During the second 6-month period, group 1 was treated with placebo, and group 2, with the same dose of vitamin C. Thirty patients in group 1 and 28 patients in group 2 completed the study. Hemoglobin levels, weekly EPO dose, and ratio of EPO to hemoglobin as an index of EPO need were determined at both baseline and the end of the two periods, together with other parameters known to be associated with EPO response. RESULTS: Twenty patients in group 1 (66.7%) and 18 patients in group 2 (64.3%) were responsive to vitamin C. In both groups, vitamin C resulted in a significant increase in hemoglobin levels (P < 0.0001 for both) and a significant decrease in EPO-hemoglobin ratio (P < 0.0001, P = 0.019). Transferrin saturation also increased with vitamin C treatment in both groups (P = 0.009, P = 0.005). All these parameters remained stable with placebo in both groups. Other parameters did not change throughout the study. CONCLUSION: Vitamin C can be used as an effective adjuvant therapy to EPO in hemodialysis patients. Further studies are needed to determine possible predictors of hematologic response to vitamin C. PMID- 12776277 TI - Hemodialysis and estrogen levels in postmenopausal (HELP) patients: The multicenter HELP study. AB - BACKGROUND: Low serum estrogen levels are associated with increased risk for hip fractures in healthy postmenopausal women. Although the rate of hip fractures is 4- to 24-fold greater in postmenopausal hemodialysis-dependent women (PHDW) compared with the general population, no published study has examined the prevalence of low serum estrogen levels or factors associated with low serum estrogen levels in this population. METHODS: We measured serum estrone (E1) and total estradiol (E2) levels in PHDW and investigated the independent association between serum estrogen levels and demographic and dialysis-related factors. RESULTS: Of 186 PHDW not administered postmenopausal hormones, 44% and 30% had low serum E1 (<15 pg/mL [55.50 pmol/L]) and total E2 levels (<5 pg/mL [18.35 pmol/L]), respectively. Serum E1 and total E2 levels were significantly lower in Caucasian women compared with non-Caucasian women and in women with a body mass index (BMI) less than 25 kg/m2 versus 25 kg/m2 or greater. After adjusting for age, BMI, dialysis vintage, dialysis dose, and history of bilateral oophorectomy, Caucasian race was associated with an increased risk for low serum E1 (odds ratio [OR], 3.42; 95% confidence interval [CI], 1.63 to 7.19) and total E2 levels (OR, 2.47; 95% CI, 1.11 to 5.49) compared with non-Caucasians. In addition, we noted a significant and independent association between BMI and serum estrogen levels. For every 1-kg/m2 increase in BMI, risk for low serum E1 and total E2 levels decreased by 12% (OR, 0.88; 95% CI, 0.83 to 0.94) and 15% (OR, 0.85; 95% CI, 0.80 to 0.91), respectively. No significant association was noted between age, dialysis vintage, or dialysis dose and low serum E1 or total E2 levels. CONCLUSION: A substantial number of PHDW have low serum estrogen levels. Caucasian race and low BMI are independently associated with low serum estrogen levels in PHDW, whereas dialysis-related factors are not. PMID- 12776278 TI - Outcome and complications of intraoperative hemodialysis during cardiopulmonary bypass with potassium-rich cardioplegia. AB - BACKGROUND: Potassium-rich cardioplegia has advantages over other cardioplegic solutions in preserving the myocardium during cardiopulmonary bypass, but it is avoided in patients with renal failure because of hyperkalemia. METHODS: We first determined the ability of intraoperative hemodialysis (IHD) to remove potassium during cardiopulmonary bypass with potassium-rich cardioplegia in 9 patients by measuring potassium levels in all dialysate and urine. We then studied 24 patients with renal failure, grouped with the 9 previous patients, to assess safety, rebound hyperkalemia, and patient outcome with this technique. RESULTS: In the first phase, 9 patients were administered 128 +/- 11 mmol of potassium in potassium-rich cardioplegia, and IHD removed 157 +/- 23 mmol. Urinary potassium excretion was only 10 +/- 3 mmol. Potassium removal occurred at a rate of 1.25 mmol/min with 0-mEq/L (mmol/L) potassium dialysate and a rate of 0.75 mmol/min with 3.0-mEq/L (mmol/L) potassium dialysate. In all 33 patients, successful initiation of cardiac rhythm occurred after cardiopulmonary bypass, and 5 patients had cardiac arrhythmias possibly from hypokalemia. In the next 24 hours, 5 dialysis-dependent patients developed hyperkalemia (potassium > 5.2 mEq/L [mmol/L]) requiring hemodialysis. Postoperative hemodialysis was delayed 2 to 3 days in the other patients. The overall death rate was 24% at 30 days. CONCLUSION: IHD effectively and safely removes potassium administered during potassium-rich cardioplegia during cardiopulmonary bypass in patients with renal failure and prevents postoperative hyperkalemia in the majority of patients. Overall mortality in patients with acute and chronic renal failure undergoing cardiac surgery is high irrespective of control of potassium balance in these patients. PMID- 12776279 TI - Plasma brain natriuretic peptide concentration on assessment of hydration status in hemodialysis patient. AB - BACKGROUND: Brain natriuretic peptide (BNP) is released into circulation in response to ventricular dilatation and pressure overload. Plasma BNP concentration correlates with left ventricular mass and dysfunction, which is prevalent in hemodialysis (HD) patients. METHODS: To evaluate the potential of BNP level for determination of hydration status, we measured inferior vena caval diameter (IVCD) and BNP levels and performed bioimpedance analysis in 49 HD patients. RESULTS: Pre-HD BNP levels remained unchanged after HD. Agreement between IVCD and pre-HD BNP level in overhydration was significant (kappa = 0.304). The area under the receiver operating characteristic (ROC) curve for overhydration was 0.819 for pre-HD BNP level. When extracellular fluid/total-body water (ECF/TBW) ratios of HD patients were compared with those of 723 controls, pre- and post-HD BNP levels were significantly greater in overhydrated patients. The area under the ROC curve for overhydration by ECF/TBW ratio was 0.781 for pre HD BNP level. However, there was no significance for pre- or post-HD BNP levels on assessment of normohydration or underhydration. Pre-HD BNP level correlated significantly with post-HD BNP level, post-HD diastolic blood pressure, pulse pressure, and ECF/TBW ratio. IVCD correlated significantly with post-HD BNP level. CONCLUSION: BNP level seems to have a limited potential for assessment of overhydration in HD patients. PMID- 12776280 TI - Heart failure as a cause for hospitalization in chronic dialysis patients. AB - BACKGROUND: Risk factors for heart failure (HF) have not been reported previously in a nationally representative sample of dialysis patients. METHODS: We conducted a historic cohort study of 1,995 patients enrolled in the US Renal Data System Dialysis Morbidity and Mortality Study Wave 2 who were Medicare eligible at the study start and were followed up until December 31, 1999, or receipt of a renal transplant. Cox regression analysis was used to model associations with time to first hospitalization for both recurrent and de novo HF (International Classification of Diseases, Ninth Revision code 428.x), defined as patients with and without a history of HF, respectively. RESULTS: The incidence density of HF was 71/1,000 person-years. Angiotensin-converting enzyme inhibitors and beta blockers were each used in less than 25% of patients with a known history of HF. A history of coronary heart disease was associated with an increased total risk for HF, as were hemodialysis (versus peritoneal dialysis), aspirin use, and a history of diabetes. However, hemodialysis and aspirin use were the only factors associated with both de novo and recurrent HF. Widened pulse pressure was associated with de novo HF. The mortality rate after HF was 83% at 3 years (adjusted hazard ratio for mortality, 2.10; 95% confidence interval, 1.80 to 2.45; P < 0.0001). CONCLUSION: In chronic dialysis patients, hemodialysis and aspirin use were associated with increased risk for both total and de novo HF. Hospitalized HF was associated with a significantly increased risk for death. PMID- 12776281 TI - Biological dynamics of hepatitis B virus load in dialysis population. AB - BACKGROUND: Control of the spread of hepatitis B virus (HBV) infection in dialysis units has been one of major advances in the management of end-stage renal disease. However, the natural history of HBV in dialysis patients remains unclear. The aim of this study is to measure monthly HBV viral load (HBV DNA) in a large cohort (n = 29) of hepatitis B surface antigen (HBsAg)-positive chronic dialysis patients during 12 months. METHODS: HBV DNA was measured using the Amplicor HBV Monitor Test (Roche Diagnostics, Branchburg, NJ), an in vitro assay using polymerase chain reaction nucleic acid amplification and DNA hybridization for the quantitative measurement of HBV DNA in serum. RESULTS: We observed three HBV DNA patterns: (1) patients persistently positive by Amplicor HBV Monitor Test (persistent HBV DNA; 7 of 29 patients; 24.1%), (2) individuals with alternatively positive and negative results (intermittent HBV DNA; 18 of 29 patients; 62.1%), and (3) patients persistently negative by Amplicor HBV Monitor Test (4 of 29 patients; 13.8%). HBV viral load was greater in patients with persistent compared with intermittent HBV DNA (persistently HBV DNA positive; 2.686 x 10(4) copies/mL; 95% confidence interval [CI], 5.2499 x 10(4) to 1.8158 x 10(4)copies/mL) versus intermittently HBV DNA positive (1.071 x 10(3) copies/mL; 95% CI, 8.524 x 10(3) to 4.09 x 10(2) copies/mL; P = 0.0001). In the entire group, HBV load at study entry was low and did not change versus the end of follow-up. CONCLUSION: Three patterns of HBV viremia in dialysis patients over time were assessed; HBV load was not high and was relatively stable. HBsAg positive patients who were intermittently HBV DNA positive had less HBV viral load than persistently HBV DNA-positive patients. Periodic testing for HBV DNA to assess the virological status of HBsAg-positive dialysis patients is recommended. PMID- 12776282 TI - Medical outcomes study short form-36: a consistent and powerful predictor of morbidity and mortality in dialysis patients. AB - BACKGROUND: One of the guidelines released by the National Kidney Foundation Kidney Disease Outcomes Quality Initiative (K/DOQI) recommends that patients with glomerular filtration rate (GFR) less than 60 mL/min/1.73 m2 undergo regular assessment of functioning and well-being (FWB) to establish baselines, monitor changes in FWB over time, and assess the effect of interventions on FWB. Although this recommendation stresses the importance of assessing and monitoring physical and mental health functioning, the Medical Outcomes Study Short Form-36 (MOS SF 36) might also be useful for predicting crucial longer-term patient outcomes. This cross-sectional study tested the hypothesis that the Physical Component Summary (PCS) and Mental Component Summary (MCS) scales of the MOS SF-36 predict morbidity (measured as hospitalization) and mortality rates among dialysis patients. METHODS: Data were collected from 13,952 prevalent dialysis patients served by Fresenius Medical Care North America including age, gender, race, diabetes, serum albumin, creatinine, bicarbonate, potassium, phosphorus, hemoglobin, iron, ferritin, white blood cell count, urea reduction ratio, serum glutamic oxaloacetic-transaminase, and systolic blood pressure. FWB was measured via the MOS SF-36 Summary scale scores, PCS, and MCS. Also collected was information about hospitalizations and patient mortality. RESULTS: PCS and MCS were consistent predictors of hospitalizations and mortality rates even after adjustment for clinically relevant factors. CONCLUSION: Because PCS and MCS are associated with hospitalization and mortality, administering this self-report measure may serve as a valuable supplement to clinical measures traditionally relied on to predict patient outcomes. Moreover, such information may be unavailable through any other single mechanism. PMID- 12776284 TI - Increased expression of p16(INK4a) and p27(Kip1) cyclin-dependent kinase inhibitor genes in aging human kidney and chronic allograft nephropathy. AB - BACKGROUND: The goal of the current study was to examine the potential value of p16(INK4a) and p27(Kip1) cyclin-dependent kinase inhibitor (CDKI) genes in the process of human kidney aging in vivo, and in the development of chronic allograft nephropathy (CAN). METHODS: Expression of p16(INK4a) and p27(Kip1) CDKI genes was evaluated and compared in 20 normal human kidney tissues of different ages (range, 21 to 80 years) and in 9 chronically rejected kidney grafts. Age dependency of marker expression was analyzed by the Pearson correlation and linear regression. RESULTS: Expression of p16 in cortical tubular (CTS) and interstitial (CIS) cells of normal kidney was age dependent (correlation coefficients: 0.608 and 0.726, 95% confidence interval [CI]: 0.227 to 0.828 and 0.417 to 0.884, respectively). Cortical tubular expression of p27 was also correlated with increasing age (0.672, 95% CI: 0.327 to 0.859). Linear regression analyses confirmed the linearity of marker relationship with age (coefficient of determination R(2):0.370, 0.452, and 0.527 for CIS p16, CTS p27, and CTS p16, respectively). The mean chronological and predicted graft ages (53 +/- 21 and 76 +/- 8.9 years, respectively) were significantly different (P = 0.0126). The glomeruli, tubules, and interstitial cells of rejected grafts expressed significantly higher levels of p16 and p27 than normal kidneys. Expression of p16 in glomerular and cortical interstitial cells was higher in grade 3 of CAN than in grade 2 (P = 0.013 and 0.004, respectively). CONCLUSION: The results of the current study show that expression of p16(INK4a) and p27(Kip1) CDKI genes is increased in cortical cells of the aging human kidney and in chronic allograft rejection, supporting the senescence theory of CAN. PMID- 12776283 TI - The relative importance of residual renal function compared with peritoneal clearance for patient survival and quality of life: an analysis of the Netherlands Cooperative Study on the Adequacy of Dialysis (NECOSAD )-2. AB - BACKGROUND: The guidelines from the US National Kidney Foundation Dialysis Outcomes Quality Initiative on peritoneal dialysis (PD) assume equivalence between the peritoneal and the renal solute clearance. The authors examined in a prospective cohort study of incident dialysis patients the relative contribution of residual renal function and peritoneal clearance to patient survival and quality of life (QoL). METHODS: The authors analyzed the longitudinal data on residual renal function, clearance by dialysis, and QoL of those patients who were treated with PD 3 months after the start of dialysis and participated in a prospective multicenter study in the Netherlands (n = 413). RESULTS: The mean age was 52 years, the mean residual glomerular filtration rate (rGFR) at 3 months was 4.1 mL/min/1.73 m2 (SD: 2.7), and the mean peritoneal creatinine clearance (pCrCl) at 3 months was 4.1 mL/min/1.73 m2 (SD: 1.1). The 2-year survival was 84%. For each mL/min/1.73 m2 increase in rGFR, a 12% reduction in mortality rate was found (relative risk of death [RR] = 0.88, P = 0.039). In contrast, no significant effect of pCrCl on patient survival was established (RR = 0.91, P = 0.47). The differential impact of rGFR and pCrCl was confirmed in an analysis on combined patient and technique survival and in an analysis on a number of generic and disease-specific dimensions of QoL. CONCLUSION: The beneficial effect of renal clearance and the absence of an effect of peritoneal clearance in the range of values common in current practice on patient outcome indicate that the 2 components of total solute clearance should not be regarded as equivalent. Higher peritoneal clearance targets do not necessarily improve patient outcome. PMID- 12776285 TI - Recurrence of focal segmental glomerulosclerosis after renal transplantation in patients with mutations of podocin. AB - BACKGROUND: Posttransplant recurrence of focal segmental glomerulosclerosis (FSGS) occurs in a relevant proportion of FSGS patients and represents an important clinical emergency. It is taken as a proof of the existence of circulating permeability plasma factor(s) that are also putative effectors of original proteinuria in these patients. Familial forms of FSGS do not recur, but the discovery of numerous patients with sporadic FSGS and mutations of podocin (NPHS2, that is actually an inherited disease) who received a renal graft require a re-evaluation of the problem. METHODS: To evaluate the incidence of posttransplant recurrence of FSGS in patients with NPHS2, the authors screened for podocin mutations in 53 patients with the clinical and pathologic stigmata of FSGS who had renal failure and who had undergone renal transplantation.Results. Twelve children were found to carry a homozygous (n9) or a heterozygous (n4) mutation of podocin and were classified, according to current criteria, as patients with inherited FSGS. In 5 patients of this group (38%), proteinuria recurred after renal graft and in 2, renal biopsy results showed recurrence of FSGS. Prerecurrence serum of 3 patients of this cohort was tested for antipodocin antibodies with indirect immuno-Western utilizing human podocyte extracts and were found negative. The rate of FSGS recurrence was comparable in non-NPHS2-FSGS children (12 of 27) and adults (3 of 13). Also clinical outcome of recurrence and response to plasmapheresis and immunosuppressors were comparable, suggesting a common mechanism. CONCLUSION: These data show a high rate of FSGS recurrence in patients with NPHS2 mutations that is comparable with idiopathic FSGS and describe the successful therapeutic approach. Recurrence of an apparently inherited disease should stimulate a critical review of the mechanisms of recurrence and of original proteinuria in these cases. PMID- 12776287 TI - Nephrotic syndrome and ARF in a diabetic patient. PMID- 12776286 TI - Diabetic muscle infarction: an unusual cause of acute limb swelling in patients on hemodialysis. AB - Acute muscle infarction in diabetic patients on dialysis has rarely been reported. We present a small case series of 4 patients with unusally "inflammatory" presentations that caused diagnostic challenges. Diabetic muscle infarction should be in the differential diagnosis of acutely swollen lower limbs in dialysis patients. PMID- 12776288 TI - Dosing darbepoetin alfa continued. PMID- 12776290 TI - Questions about left internal mammary artery coronary "steal". PMID- 12776291 TI - Orexin as a possible cause of insomnia in dialysis patients. PMID- 12776302 TI - Quiz page. Diabetic nephropathy with superimposed acute interstitial nephritis. PMID- 12776303 TI - Culture-negative peritoneal dialysis peritonitis associated with pancreaticoduodenocystotomy leak of a pancreas transplant. AB - BACKGROUND: Peritoneal dialysis (PD) peritonitis is usually caused by infection and less commonly by a sterile inflammatory reaction. METHODS: The authors report the case of a kidney-pancreas transplant recipient who was receiving PD after kidney transplant rejection 5 years after transplantation. The patient had a viable pancreas transplant. He had abdominal pain associated with cloudy PD effluent. The PD leukocyte count was elevated with a predominance of monocytic leukocytes. RESULTS: Blood, urine, and PD effluent cultures were negative. An ultrasound scan of the transplanted kidney and a computerized tomography (CT) scan of the abdomen and pelvis did not identify the cause of the peritonitis. Foley catheter decompression of the bladder resulted in improvement of the abdominal pain and PD effluent leukocytosis. Twenty-five days later, the patient again experienced abdominal pain and cloudy PD effluent. Cultures of blood and PD effluent were again negative. CT scanning and cystoscopy of the transplanted pancreas identified a leak at the pancreaticoduodenocystotomy anastamosis. Urinary bladder decompression was followed by surgical exploration that identified an erosion of the distal transplanted duodenum, necessitating enteric diversion of the transplanted pancreas's exocrine secretions. The patient underwent conversion to hemodialysis, and the pancreas transplant continued to function well. He has subsequently received a living related kidney transplant. CONCLUSION: This is the first reported case of noninfectious PD peritonitis caused by pancreaticoduodenocystotomy leak in a patient with a functional pancreas transplant. PMID- 12776304 TI - Delayed angioplasty after renal thrombosis. AB - When renal arterial thrombosis occurs, the etiologic process plays an important role in the impact of ischemia on renal tissue. If the occlusion is caused by trauma, infarction rapidly occurs. However, when renal arterial thrombosis results from other processes, collateral vessels may develop and thus enables a prolonged ischemia without necrosis. The following is a case report of an acute renal failure caused by renal arterial thrombosis of a single functional kidney, which had a favorable outcome despite delayed treatment by percutaneous angioplasty. This report suggests that detection by ultrasonography of a venous renal flow could be of significant value to assess a collateral vascularization and thus should require an arteriography with angioplasty even after a delayed presentation. PMID- 12776305 TI - Renal parenchymal malacoplakia: a rare cause of ARF with a review of recent literature. AB - Renal parenchymal malacoplakia is a rare cause of acute renal failure. Traditionally, it was associated with a high mortality rate and commonly resulted in renal failure requiring renal replacement therapy. The authors report on a 70 year-old woman who presented with acute renal failure caused by renal parenchymal malacoplakia. Her renal function recovered after levofloxacin treatment. All cases reported in the English-language literature since 1990, when fluoroquinolone was first used to treat malacoplakia, were reviewed. Although some patients still had renal failure, with renal biopsy and fluoroquinolone treatment, the patient mortality rate from renal parenchymal malacoplakia is remarkably low. PMID- 12776306 TI - Diagnosis of renal and hepatic cyst infections by 18-F-fluorodeoxyglucose positron emission tomography in autosomal dominant polycystic kidney disease. AB - BACKGROUND: Infection of a renal or hepatic cyst is a serious complication of autosomal dominant polycystic kidney disease (ADPKD). Although crucial for successful management, early diagnosis is difficult, largely because of nonspecific symptoms and limitations of conventional imaging techniques. Because of an increased metabolic rate, inflammatory cells take up large amounts of glucose. 18-F-fluorodeoxyglucose (FDG), therefore, represents a promising agent for detection of cyst infections using positron emission tomography (PET). METHODS: The authors studied the results of 7 FDG PET scans in 3 ADPKD patients suspected of renal or hepatic cyst infection. Two PET scans were performed in patient A (PET 1 and 2), one PET scan was performed in patient B (PET 3), and 4 PET scans were performed in patient C (PET 4, 5, 6 and 7). RESULTS: FDG PET identified the infected cysts in 2 episodes of renal cyst infection (PET 2 and 3), 2 episodes of hepatic cyst infection (PET 6 and 7), and 1 episode of both renal and hepatic cyst infection (PET 1). In patient C, FDG PET was normal after 6 weeks of antibiotic treatment for hepatic cyst infection (PET 4) and again at a time when hepatic cyst infection was suspected, but eventually colchicine intoxication was diagnosed (PET 5). CONCLUSION: In these patients, FDG PET proved very helpful in diagnosing and in excluding renal and hepatic cyst infections. It is concluded that FDG PET is a promising new imaging technique enabling early identification of renal and hepatic cyst infections in ADPKD patients. PMID- 12776308 TI - Meeting the challenges of the new K/DOQI guidelines. AB - Substantial gains in the dialytic treatment of patients with end-stage renal disease have been made during the past several decades. However, inadequate attention has been given to the problem of chronic kidney disease (CKD) as a whole. CKD and its associated complications emerge years before patients develop kidney failure and become dialysis dependent. It now is evident that to improve dialysis outcomes, it is essential for practitioners to recognize the earlier stages of CKD, not only to retard disease progression, but also to prevent and treat its complications and comorbidities long before the need for dialysis arises. The recently published National Kidney Foundation Kidney Disease Outcomes Quality Initiative guidelines identify the broad-based problem of CKD in the general population and introduce action plans that can be used at the different stages of CKD. This article discusses the process behind the development of the CKD guidelines and highlights the disease evaluation, classification, and stratification system provided by the guidelines. It is hoped that widespread implementation of these recommendations will increase the understanding of CKD among both providers and patients and eliminate many of the obstacles that nephrologists have faced in providing optimal care to dialysis patients in particular and to patients with CKD in general. PMID- 12776309 TI - Cardiovascular complications in chronic kidney disease. AB - The risk for cardiovascular disease (CVD) morbidity and mortality remains alarmingly high in all stages of chronic kidney disease (CKD). CVD often begins before end-stage renal disease (ESRD), and patients with reduced kidney function are more likely to die of CVD than to develop ESRD. Three pathological forms of CVD should be considered in patients with CKD: alterations in cardiac geometry, including left ventricular hypertrophy, atherosclerosis, and arteriosclerosis. All are highly prevalent in patients with CKD. Although patients with CKD share many of the same risk factors for CVD as the general population, there are a number of uremia-related risk factors, such as anemia and alterations in calcium/phosphorus metabolism, that also play a role in promoting CVD. Treatment of both traditional and uremia-related risk factors should be initiated in the earlier stages of CKD. Additional clinical trials with a goal to reduce CVD are urgently needed in CKD. PMID- 12776310 TI - Safety in iron management. AB - Intravenous (IV) iron therapy has become an integral part of hemodialysis management during the past several decades, and the National Kidney Foundation Kidney Disease Outcomes Quality Initiative guidelines recognize that most patients undergoing hemodialysis will require IV iron therapy on a regular basis to reach target hemoglobin (Hgb) levels. There now are three IV iron compounds available in the United States: iron dextran, sodium ferric gluconate, and iron sucrose. Although all have been proven effective for increasing Hgb/hematocrit levels, recent data show differences in their relative safety profiles. During the past two decades, more than 30 deaths have been attributed to the use of IV iron dextran. The two newer compounds available in the United States, sodium ferric gluconate and iron sucrose, have more favorable safety profiles, with the largest prospective safety comparison to date showing sodium ferric gluconate to be similar to placebo in the incidence of serious anaphylactoid-type reactions. This article reviews safety data surrounding the IV iron therapies. PMID- 12776314 TI - [Studies on original plant of traditional Chinese drug "bai zhi" (radix Angelicae Dahuricae) and its closely related wild plants. III. Comparison of coumarins of "bai zhi" with those of closely related wild plants]. AB - OBJECTIVE: To provide chemical data for confirming the original plant of traditional Chinese drug "Bai Zhi". METHOD: Coumarins of 4 cultivated breeds of "Bai Zhi" and 3 closely related wild plants, together with other 2 Angelica plants were compared by HPLC. RESULT: According to coumarin patterns, 4 cultivated breeds of "Bai Zhi" and 3 closely related wild plants could be divided into 3 groups: 1. 4 cultivated breeds of "Bai Zhi" ("Chuan Bai Zhi", "Hang Bai Zhi", "Qi Bai Zhi" and "Yu Bai Zhi") and Angelica dahurica var. formosana; 2. A. dahurica; 3. A. porphyrocaulis. CONCLUSION: In point of the coumarin components, A. dahurica var. formosana is closer to traditional Chinese drug "Bai Zhi" than the others. PMID- 12776315 TI - [Studies on morphology and histology of medicinal part of Corydalis yanhusuo]. AB - OBJECTIVE: To Study the morphology, microstructure and morphosis of medicinal part of Corydalis yanhusuo, and provide a basis for the knowledge of the type of modified stem of Corydalis yanhusuo and the identification of crude drug. METHOD: Sample collection, culture observation and microsectile observation. RESULT: Morphological and histological characters and morphogenetic and developmental regulation of medicinal part of Corydalis yanhusuo are described with pictures. CONCLUSION: The medicinal part of Corydalis yanhusuo is bulb, and the conception that the medicinal part of Corydalis yanhusuo is tuber is wrong. PMID- 12776316 TI - [Two new glycosides from Erigeron breviscapus (Vant.) Hand.-Mazz]. AB - OBJECTIVE: To study the chemical constituents from the upground part of Erigeron breviscapus. METHOD: The compounds were separated and purified by column chromatography with silica gel, and identified by IR, MS, NMR and 2D-NMR. RESULT: Two new compounds were isolated and identified as 5,4'-dihydroxy flavonod-7-O beta-D-pyranglycuronate buthyl ester(VI) and 3,5-dimethoxy benzene carbonic acid 4-O-beta-D-pyranglucose(VII). CONCLUSION: Compounds VI and VII were new compounds. PMID- 12776317 TI - [Studies on chemical constituents of Bletilla striata (Thunb) Reichb. f]. AB - OBJECTIVE: To study the chemical constituents of Bletilla striata. METHOD: The constituents were separated and purified by column chromatography with silica gel, and identified by NMR, MS and physical data. RESULT: Three compounds were isolated and identified as hexacosanoic alcohol 3-(4-hydroxy-3-methoxybenzol) trans-acryliceylenate(1), physcion(2) and cyclobalanol(3). CONCLUSION: Compound 1 was a new compound and compound 3 was isolated from this plant for the first time. PMID- 12776318 TI - [Chemical constituents of Loxocalyx urticifolius Hemsl]. AB - OBJECTIVE: To investigate the chemical constituents from the whole plant of Loxocalyx urticifolius. METHOD: The chemical constituents were isolated by chromatography and identified by MS, 1H NMR, 13C NMR and 2D-NMR methods. RESULT: The structures were established as poliothrysoside, beta-sitosterol and stigmasterol, palmitic acid and tyrosine. CONCLUSION: This is the first time for the chemical constituents of L. urticifolius to be reported. PMID- 12776319 TI - [Studies on chemical constituents of Schisandra propinqua (Wall.) Hook. f. et Thoms]. AB - OBJECTIVE: To isolate and characterize compounds from the stems of Schisandra propinqua. METHOD: Extracting with solvent, isolating by column chromatography and identifying by the spectroscopic methods. RESULT: Six dibenzocyclooctadiene lignans were isolated and identified as tigloylgomisin P(1), angeloylgomisin O(2), angeloylisogomisin O(3), kadsulignan L(4), (+/-) 5,8-epoxyl-6, 7-dimethyl 2',3',2",3"-dimethylenedioxy-4', 1"-dimethyl-1,2:3,4-dibenzo-1, 3 cyclooctadiene(5) and wuweizisu C(6). CONCLUSION: Compounds 4 and 5 were the first two dibenzocyclooctadiene lignans with an 6,9-epoxy bridge cycle discovered in the genus Schisandra. The others were originally isolated from S. propinqua. PMID- 12776320 TI - [Studies on chemical constituents from Dracocephalum moldavica L]. AB - OBJECTIVE: To study the chemical constituents from the whole plant of Dracocephalum moldavica. METHOD: The compounds were isolated using RA polystyrene resin and silica gel column chromatography, and the structures were elucidated by means of spectral method. RESULT: Four compounds were identified as tilianin, agastachoside, acacetin and oleanolic acid. CONCLUSION: All the compounds were isolated from this plant for the first time. PMID- 12776321 TI - [A comparison of chemical composition and bioactivity of polypeptides from velvet antlers of Cervus nippon Temminck and Cervus elaphus Linnaeus]. AB - OBJECTIVE: To compare the chemical composition and bioactivity of polypeptides(PPs) isolated from velvet antlers of sika deer (Cervus nippon Temminck) and red deer (Cervus elaphus Linnaeus). METHOD: The two kind of polypeptides were isolated from the above mentioned velvet antlers with same technology. The chemical composition was determined using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and MALDI-TOF mass spectrometry. Stimulant activity of cells proliferation was measured by [3H] TdR incorporation into DNA. RESULT: The graphs of SDS-PAGE and MALDI-TOF MS of velvet antler polypeptides (VAPPs) from Chinese and New zealand red deer were very similar, but there were obvious difference in respect of graph between sika deer and red deer. VAPPs 25-50 mg.L-1 showed marked proliferation-promoting activity for rabbit costed chondrocytes, either sika deer or red deer. However, the activity of sika deer VAPPs 12.5 mg.L-1 for epidermal cells was weaker than that of red deer (12.5 mg.L-1). CONCLUSION: The chemical property and bioactivity of VAPPs from sika deer and red deer are significantly different. PMID- 12776322 TI - [Study of protective effect of leech, radix Salviae miltiorrhizae and its composite recipe on vascular endothelial cells in rats with blood stasis syndrome]. AB - OBJECTIVE: To study the protective effect for Chinese medicine of promoting blood circulation and removing blood stasis (PBCRBS) on vascular endothelial cells (VEC) in rats, aiming at further research on the mechanism of blood stasis syndrome and PBCRBS. METHOD: Establishing a model of blood stasis with endothelial damage by means of giving rats an injection of adrenalin and making it swim in ice-cold water, then measuring the number of circulating endothelial cells (CEC) in whole blood and rheology. Moreover observing the change of the above indexes after the rats have taken Leech, Radix salviae miltrorrhizae (RSM) and its composite recipe for 5 days or 10 days. RESULT: It could increase the number of CEC accompanied by whole blood viscosity, fibrinogen and hematocrit in rats with blood stasis. But for groups taking Chinese medicine, the number of CEC was decreased distinctively and blood rheology was improved. CONCLUSION: The rat with blood stasis syndrome suffered from obvious injury of VEC. The Leech, RSM and its composite recipe could protect VEC. PMID- 12776323 TI - [Experimental studies of antitumor effect of artesunate on liver cancer]. AB - OBJECTIVE: To observe the inhibiting effect of Artesunate on liver cancer in vitro and in vivo. METHOD: The mice bearing H22 solid and ascitic liver tumor were applied in vivo experiments. Microculture tetrazolium assay and colony forming unit assay were applied to test the cytotoxicity to human hepatocarcinoma SMMC-7721 cell line in vitro. RESULT: The growth of solid tumor were obviously inhibited by Artesunate at the dose of 300 mg.kg-1.d-1 ig for 7 days. The tumor inhibiting rates of Artesunate were 49.1%, 48.7%, 46.6% in 3 experiments respectively. After administration of Artesunate, the survival rate of the mice bearing H22 ascitic liver tumor were increased to 45%. Compared with the control groups, the difference was statistically significant (P < 0.01). In additional, Artesunate can synergize the antitumor activity of 5-fluorouracil. Artesunate showed evident cytotoxicity to human hepatocarcinoma SMMC-7721 cells, the IC50 of Artesunate being 2.07 micrograms.ml-1 in MTT experiment and 2.48 micrograms.ml-1 in colony-forming unit experiment. CONCLUSION: Artesunate has marked antitumor activity in vitro and in vivo. PMID- 12776324 TI - [Studies on polysaccharide alkaloids and minerals from Dendrobium moniliforme (L.) Sw]. AB - OBJECTIVE: To explore contents of active substances in different part of Dendrobium monilifrome and the quality influenced by different drying processes. METHOD: Contents of alkaloids and polysaccharides in upper, middle, lower-stem and root part of Dendrobium moniliforme were determined by uv-spectrophotometer method. Contents of essential trace elements were determined by ICP method. RESULT: Contents of polysaccharides and alkaloids were highest in the upper-stem of D. montiliforme. Contents of active substances in different part of D. moniliforme were unequal. Distributions of polysaccharides and alkaloids in different parts were greatly influenced by drying procedures. CONCLUSION: Dendrobium moniliforme has higher contents of active ingredients, such as essential tract elements, alkaloids and polysaccharides. The ideal drying process after harvesting is to be the way dried by fire at a high temperature and desiccated at a temperature of 60 degreeC. PMID- 12776325 TI - [Comprehensive utilization and development of traditional Tibetan medicine in China]. AB - OBJECTIVE: To deal with the further investigation field by discussing the status and present problem of traditional Tibetan medicine. METHOD: Previous relevant investigations and literatures were summed up in the field. The present situation of traditional Tibetan medicine in China was analysed. RESULT: The textual research, basic medicinal property, exploration of developable medicinal resource and protection of endangered medicinal species etc. were elaborated and the key problem of further investigation in 21st century was expounded. CONCLUSION: The textual research, basic medicinal property, exploration of develoable medicinal resources, especially monographic study on protection of major endangered medicinal resources should be intensified. Domestication and cultivation, and exploration of good-quality medicinal resources, quality evaluation and exploitation of effectual prescriptions are the focal field in the study of traditional Tibetan medicine. PMID- 12776326 TI - [Induction of callus of Hypericum perforatum L. and qualitative identification of active constituents of its callus]. AB - OBJECTIVE: To induce the callus of H. perforatum and identify hypericin and pseudohypericin of its callus. METHOD: The callus was induced in different culture conditions, and active constituents were determined by HPLC. RESULT AND CONCLUSION: The inductions of callus from different parts were discussed, the induction rate of the leaf axil being the highest. The MS basic medium with 4 micrograms.L-1 2,4-D and 0.2 microgram.L-1 6-BA was the best of all screened media. Hypericin in the callus is determined by HPLC. PMID- 12776327 TI - [The microscopic observed for cells in mesocarp of part medicinal plant in Citrus L]. AB - OBJECTIVE: To illustrate the microscopic characters of cells in mesocap of some of medicinal plants in Citrus of Rutaceae. METHOD: Microscopic observation and photograph were carried out. RESULT: It has been found that many of pavenchymatous cells are branches. It is spongy tissue that has huge room between cells. The thickening of cell wall is not obvious. The uneven thickness of walls of cells in mesocarp which is reported in the old documents are not found. PMID- 12776328 TI - [Studies on the arbutin biosynthesis by hairy root of Panax ginseng C.A. Mayer]. AB - OBJECTIVE: To study basic conditions for biosynthesis arbutin by the hairy root of Panax ginseng. METHOD: Based on the content of arbutin and the biotransformation rate of hydroquione, these conditions such as the culture stage, the lasting time and the concentration of hydroquione fitting for biosynthesis were found. RESULT: After cultured for 22 days, transferred in fresh B5 medium having 2 mmol.L-1 hydroquione, the hairy root biotransformated hydroquione into arbutin in 24 hours, the bioconversion rate was 89.0% and the content of arbutin in dry root was 13.0%. CONCLUSION: Arbutin, was which never found in the genus of Panax ginseng, can be biosynthesized by the hairy root of Panax ginseng. PMID- 12776330 TI - [Constituents of Liguliria vellerea (Franch.) Hand-Mazz]. AB - OBJECTIVE: To study the chemical constituents of Ligularia vellerea. METHOD: The compounds were isolated by column chromatography, and the structures were identified by NMR spectral data and other methods. RESULT AND CONCLUSION: Seven compounds were isolated and identified as 4-hydroxyacetophenone, 8 alpha-hydroxy 7(11)-eremophilen-12, 8 beta-olide, umbelliferone, tiglic acid, 6 beta-hydroxy 7(11)-eremophilen-12, 8 alpha-olide, daucosterin, beta-sitosterol and stigmasterol. All the compounds were isolated for the first time from this plant. PMID- 12776329 TI - [A comparative study on sterols of ethanol extract and water extract from Hericium erinaceus]. AB - OBJECTIVE: To study the pharmaceutic chemical basis of the different medicinal effects, and to compare and analyze the sterols in Hericium erinaceus mycelia and its ethanol extract, water extract derived from solid fermented mycelia. METHOD: The components of Hericium erinaceus mycelia and two kinds of extract were compared with some biochemical methods such as GC, RP-HPLC, etc. Sterol composition in Hericium erinaceus mycelia was determined by using GC-mass spectrometry. RESULTS: The content of crude polysaccharide, water-soluble protein and fatty acid of ethanol extract showed no obvious advantage over its water extract. However, there was significant difference between two kinds of extract in the content of sterols. And ergostane derivatives, beta-sitosterol and C28 sterol with four bonds were detected as the major sterols in Hericium erinaceus mycelia, among which ergosterol was the principal sterol. CONCLUSION: Sterols in Hericium erinaceus exist mainly in ethanol extract. And three sterols are found for the first time from Hericium erinaceus, which are ergostane derivatives, beta sitosterol and C28 sterol with four bonds. PMID- 12776331 TI - [Studies on the chemical constituents of Carduus crispus L]. AB - OBJECTIVE: To study the chemical constituents of Carduus crispus. METHOD: Using chromatographic methods to isolate compounds and using chemical and spectral methods to elucidate the structures of the isolated compounds. RESULT: Eight compounds were elucidated as beta-amyrin palmitate, taraxastery acetate, luteolin 7-O-alpha-L-rhamanopyranosyl-(1-->2)-beta-D-glucopyranoside, luteolin-7-O-beta-D glucopyranoside, triacontanic acid, beta-sitosterol, stigmasterol and stigmast-7 en-3 beta-ol. CONCLUSION: All the compounds were obtained from this plant for the first time. PMID- 12776332 TI - [Two kaempferol triglycosides from pericarps of Sophora japonica L]. AB - OBJECTIVE: To study the flavonol triglycosides in the pericarps of Sophora japonica. METHOD: Various chromatographic techniques were used to isolate and purify the constituents. The structures were elucidated by chemical evidence and spectral analysis, especially by 2D NMR experiments. RESULTS: Two kaempferol triglycosides were isolated and identified as kaempferol 3-O-beta-D-sophoroside-7 O-alpha-L-rhamnoside and kaempferol 3-O-(2"-O-beta-D-glucosyl)-beta-D-rutinoside. CONCLUSION: Both of them were reported in S. japonica for the first time. PMID- 12776333 TI - [Studies on flavonoids from leaves of Callicarpa bodinieri Levl]. AB - OBJECTIVE: To study the constituents of Callicarpa bodinieri. METHOD: The compounds were isolated by chromatography. Their structures were identified by physical and spectral data. RESULT: Seven compounds were isolated and elucidated as 5-hydroxy-4',3,6,7-tetramethoxy flavone(I), luteolin-7-O-glucoside (II), chrysoeriol-4'-O-glucoside(III), luteolin-4'-O-glucoside(IV), beta-sitosterol(V), ursolic acid(VI), betulinic acid(VII). CONCLUSION: All the compounds were isolated from this plant for the first time; Compounds I, III, IV were isolated from Callicarpa genus for the first time. PMID- 12776334 TI - [Studies on chemical constituents of Gaultheria leucocarpa var. Yunnanensis (Franch.) T. Z. Hsu & R. C. Fang]. AB - OBJECTIVE: To separate and identify the chemical constituents of the aerial part of Gaultheria leucocarpa var. yunnanensis. METHOD: The compounds were extracted with solvents, isolated by column chromatography and identified by spectral analysis. RESULT: Four compounds were identified as n-dotriacontane and its homologous compound(1), ursolic acid(2), vanillic acid(3), and quercitrin(4). CONCLUSION: The compounds 1, 4 were obtained from the plant for the first time, and 2 and 3 were from above-ground part of the plant for the first time. PMID- 12776335 TI - [Determination of evodiamine and rutaecarpine in Evodia rutaecarpa (Juss.) Benth after compatibility with Coptis chinensis Franch]. AB - OBJECTIVE: To studying changes of evodiamine and rutaecarpine in E. rutaecarpa after compatibility with C. chinensis. METHOD: An YWG C18 column was used with a mobile phase of CH3CN-H2O(40:60), the flow rate was 1.0 ml.min-1 and the detection wavelength was 290 nm. RESULT: The average recovery was 97.82% and 97.00%, the relative standard deviation was 0.81% and 1.10% for evodiamine and rutaecarpine respectively. CONCLUSION: The amounts of evodiamine and rutaecarpine were decreased after compatibility with C. chinensis. PMID- 12776336 TI - [Determination of salvianolic acid B in the radix of Salvia miltiorrhiza Bge. by HPLC]. AB - OBJECTIVE: To develop an HPLC method for determining salvianolic acid B in radix of Salvia miltiorrhiza. METHOD: The sample was extracted with 75% methanol. ODS column was used with methanol-5% acetic acid solution (35:65) as mobile phase. Detection wavelength was 281 nm. RESULT: Salvianolic acid B was separated well. Good linearity of salvianolic acid B was obtained (r = 0.9997) within the range of 0.17-1.7 micrograms. The average recovery was 98.9%. Repeatability was good, and RSD was 1.82%. CONCLUSION: This method can be used for quality control of radix of Salvia miltiorrhiza. PMID- 12776337 TI - [Effect of satragaloside IV on the increase of microvascular permeability induced by histamine in pial microvessels of rat]. AB - OBJECTIVE: To study the effect of satragaloside IV on the microvascular permeability induced by histamine in pial microvessels. METHOD: The microvascular permeability was expressed by changes in the transendothelial electrical resistance which was measured with technique using microelectrode impaled into the vascular lumen and based on cable analysis of vessels in rat. RESULT: The transendothelial electrical resistance of microvessels superfused with artificial cerebrospinal fluid was about 2500 omega.cm2, indicating a tight barrier with extremely low ion permeability, and application of 10(-4) mol.L-1 histamine in superfusate caused a rapid and reversible decrease in transendothelial electrical resistance. In paired experiment, the decrease of transendothelial electrical resistance induced by 10(-4) mol.L-1 histamine was inhibited by adding 0.8 x 10( 4) mol.L-1 satragaloside IV in superfusate. CONCLUSION: The results indicated that increases in the microvascular permeability induced by histamine, and satragaloside IV can inhibit the increases in the microvascular permeability induced by histamine. It is necessary that the cellular mechanism of permeability response induced by satragaloside IV be further elucidated. PMID- 12776338 TI - [The effect of kudingcha (Ilex latifolia Thunb.) on the contraction of isolated guinea-pig tracheal smooth muscle in vitro]. AB - OBJECTIVE: To study the effects of Kudingcha (Ilex latifolia Thunb., IL) on the isolated guinea pig tracheal smooth muscle in vitro. METHOD: By obtaining CaCl2 and Histamine accumulative dose-response curves, to observe the influences of IL on the contraction of tracheal strips induced by calcium and some asthmogenic mediators. RESULT: CaCl2 and histamine caused significant contraction of tracheal smooth muscle and pD2 was 3.55 and 5.34 respectively. After incubated with IL, the dose-response curves of CaCl2 and histamine were significantly shifted to the right, and the maximal contractile force was reduced. IL could also inhibit isolated tracheal strip contraction induced by acetylcholine 3 x 10(-6) mol.L-1 and histamine 3 x 10(-6) mol.L-1, and IC50 was 0.16 mg.ml-1 and 0.21 mg.ml-1. CONCLUSION: Kudingcha has significant dilated effects on tracheal smooth muscle. PMID- 12776339 TI - [Inhibitive effect of puerarin on increased NO production by neonatal cardiomyocytes during hypoxia/reoxygenation injury]. AB - OBJECTIVE: To investigate the effect of puerarin (Pue) on nitric oxide (NO) produced by neonatal rat cardiomyocytes during hypoxia/reoxygenation injury. METHOD: NO contents in the culture supernatants sampled from different groups (control, model, and therapeutic groups with 1, 0.1, 0.01 g.L-1 Pue) at different time were assayed with nitrate reductase method. RESULT: NO content of model increased after reperfusion (P < 0.01), while it increased sharply at 6 h after reperfusion and kept thereafter. Only at 6 h after reperfusion did Pue in these three doses inhibit NO production (P < 0.01) and kept to the 24 h after reperfusion. CONCLUSION: Pue exerts inhibitive effect only after NO production is enhanced sharply during hypoxia/reperfusion injury in a dose-dependent trend. PMID- 12776340 TI - [Effect of alcohol extract of Cornus officinalis Sieb. et Zucc on GLUT4 expression in skeletal muscle in type 2 (non-insulin-dependent) diabetic mellitus rats]. AB - OBJECTIVE: Based on its effects of decreasing postprandial plasma glucose and increasing insulin level in non-insulin-dependent diabetic mellitus (NIDDM) rats, we studied the effects of Alcohol extract of Cornus officinalis Sieb. et Zucc on the GLUT4 expression in NIDDM model rats. METHOD: The rat model of NIDDM was made. The animals were divided into three groups(six for each group): group I: control; group II: NIDDM model; group III: NIDDM model + Cornus officinalis Sieb. et Zucc. The drug was given orally to animals one time a day, uninterrupted for a month. The GLUT4 mRNA and its protein expression in skeletal muscle were observed with Northern blot and Western blot method, respectively. RESULT: The GLUT4 mRNA expression in skeletal muscle was decreased remarkably in NIDDM rats (P < 0.01, compared to control group). Alcohol extract of Cornus officinalis Sieb. et Zucc increased GLUT4 mRNA expression in NIDDM rats(P < 0.01), compared to NIDDM model group). The result of GLUT4 protein expression was similar to GLUT4 mRNA expression. CONCLUSION: This experiment demonstrated that alcohol extract of Cornus officinalis Sieb. et Zucc can increase GLUT4 mRNA and its protein expression in NIDDM rats through promoting proliferation of islet and increasing postprandial secretion of insulin and therefore accelerate glucose transport. PMID- 12776341 TI - [System study of endangerment medicinal animals in China]. AB - OBJECTIVE: Establishing system study of endangerment medicinal animals is essential for the protection of certain species of medicinal wild fauna. METHOD: It was analyzed and summed up according to the study and protection actuality of endangerment medicinal animal in recent years. RESULT AND CONCLUSION: It was essential to research endangerment medicinal animals for realizing modernization, industrialization, internationalization and stabilization development of Chinese Materia Medica. PMID- 12776342 TI - [Studies on original plant of traditional Chinese drug "bai zhi" (radix Angelicae Dahuricae) and its closely related wild plants. IV. Discussion on original plant and cultivation history of traditional Chinese drug "bai zhi" and evolution of its closely related wild plants]. AB - OBJECTIVE: To confirm the original plant of traditional Chinese drug "Bai Zhi" and to inquire into the cultivation history of "Bai Zhi" and evolution of closely related wild plants of "Bai Zhi". METHOD: Various research results obtained were synthesized and discussed according to historical and current data. RESULT: Obtained research results, historical and current data showed almost no difference. CONCLUSION: 1. Angelica dahurica var. formosana must be the original plant of traditional Chinese drug "Bai Zhi". 2. A. porphyrocaulis should be treated as a variety of A. dahurica, named as A. dahurica var. porphyrocaulis. 3. 4 sorts of Chinese traditional drug "Bai Zhi" (Chuang Bai Zhi, Hang Bai Zhi, Qi Bai Zhi and Yu Bai Zhi) should not be taxonomically distinguished. The history of utilization and cultivation of "Bai Zhi", and the evolutional relation of the closely related wild plants of "Bai Zhi" (A. dahurica, A. dahurica var. formosana, and A. dahurica var. porphyrocaulis) were also discussed. PMID- 12776343 TI - [Determination of content of nodakenin in Notopterygium incisum ting from different source by HPLC]. AB - OBJECTIVE: To develop a method for the determination of nodakenin in N. incisum. METHOD: A HPLC method was used. RESULT: Y = -9509.1 + 1912709.8X, r = 0.9998. The calibration was linear in range of 0.16-0.64 microgram. The average recovery was 99.7%, RSD 1.1%. CONCLUSION: The method is appropriate for the determination of nodakenin. PMID- 12776344 TI - [Selection of high taxol content cell lines of Taxus yunnanensis Cheng et L. K. Fu]. AB - OBJECTIVE: To select high taxol content cell lines of Taxus yunnanensis. METHOD: Choosing different cell aggregates according to their color, texture, growth rate and secretion of colorful substances, culturing them separately, and further analyzing their growth rates and taxolcontents. RESULT AND CONCLUSION: Cell lines with higher taxol contents could be obtained by careful selection; those with darker color, lower growth rate and higher ratio of dry cell weight vs. fresh cell weight usually has higher taxol contents. PMID- 12776345 TI - [Study on supercritical CO2 fluid extraction and separation of components from fruits of Schisandra sphenanthera]. AB - OBJECTIVE: To extract and separate deoxyschisandrin, gamma-schisandrin, etc in fruits of Schisandra sphenanthera by supercritical carbon dioxide fluid extraction technology. METHOD: Optimum extraction conditions were studied by orthogonal tests and deoxyschisandrin, gamma-schisandrin, wuweizi chun A, wuweizi ester A were analyzed by HPLC. RESULT AND CONCLUSION: The optimum extraction conditions are pressure 21 MPa, temperature 37 degrees C and flow rate of CO2 5 L.min-1, according to the rate of extraction. PMID- 12776346 TI - [Determination of puerarin in xinmaitong oral liquid by HPLC]. AB - OBJECTIVE: To develop a method to determine the content of puerarin in Xinmaitong oral liquid. METHODS: HPLC method was used. The separation was performed on Suntek kromasil C18 column with methanol-water containing 0.5% acetic acid (25:75) as a mobile phase and the wavelength of UV detector was 250 nm. RESULTS: The linearity of puerarin was good and average recovery was 97.8%. CONCLUSION: The method is simple, reliable and sensitive. It also shows good resolation. It can be used in quality control of Xinmaitong oral liquid. PMID- 12776347 TI - [Studies on the chemical constituents of Asarum longerhizomatosum C. F. Liang et C. S. Yang]. AB - OBJECTIVE: To study the chemical constituents of the roots and rhizomes of Asarum longerhizomatosum. METHOD: Chromatography and spectral analysis were used to isolate the constituents and elucidate their structures. RESULT: Five compounds were isolated from the ethanol extracts of the roots and rhizomes, and identified as asarone(I), beta-sitosterol(II), 2,4,5-trimethoxybenzaldehyde(III), 4-(2,4,5 trimethoxyphenyl)-3-en-butylone(IV) and 3 beta-hydroxystigmast-5-en-7-one(V). CONCLUSION: All the compounds were isolated from the plant for the first time, and IV is a new natural product. PMID- 12776348 TI - [Structure determination of the constituents from Citrus grandis Osbeck]. AB - OBJECTIVE: To isolate the chemical constituents of the peelings of Citrus grandis and determine their structure. METHOD: Column chromatography and PTLC were applied to isolating and purifying the chemical constituents, and 1H NMR and 13C NMR were used to elucidate their structure. RESULT: 4 compounds were isolated as meranzin hydrate(1), roseoside(2), 8-(3-beta-D-syl-2-hydrox-3-methylbutyl)-7 methoxycoumarin(3) and glucose(4). CONCLUSION: Compound 2 was isolated as a carotenoid from this genus for the first time. PMID- 12776349 TI - [Studies on constituents of the leaves of Lindera aggregata (Sims) Kosterm]. AB - OBJECTIVE: To study the antibacterial and anti-inflammatory constituents of the leaves of Lindera aggrega. METHOD: Compounds were isolated by colum chromatography, and the structures were identified by spectroscopic methods. RESULT: Six compounds were isolated and identified as mixture of 6 Acetyllindenanolide B-1 and B-2(I), dehydrolindestrenolide (II), hydroxylinderstrenolide (III), linderalactone (IV), kameofero (V), beta sitosterol (VI). CONCLUSION: These compounds were obtained from the leaves of Lindera aggregata for the first time. PMID- 12776350 TI - [Studies on metabolites of baicalin in human urine]. AB - OBJECTIVE: We are interested in the metabolism of baicalin in human body to offer more information to clinic. METHOD: Urine of ten persons was collected during 2 18 h after taking 1.0 g of baicalin orally, and the urine was concentrated to 0.5 L, then filtered, the solution being loaded on an iron exchange column eluted with 1.0 L of H2O and 1.0 L of 50% EtOH. The EtOH fraction was concentrated and loaded on a Sephadex LH 20 column eluted with 0.5 L of H2O to obtain 15 fractions, which were detected by silica gel TLC (CHCl3-MeOH-H2O, 1:1:0.1). The fractions 8-11 showed flavonoid characteristics by spraying FeCl3 reagent, and the four fractions were combined to obtain the sample containing metabolites of baicalin for HPLC analysis and LCMS measurement. RESULTS AND CONCLUSIONS: M1, M2 and M3 was identified as baicalein-6-O-beta-glucopyranuronoside; 6-O-methyl baicalin-7-O-beta-glucopyranuronoside and baicalin. PMID- 12776351 TI - [Inhibitory effect of artemin on endotoxin-induced nitric oxide synthesis]. AB - OBJECTIVE: To investigate the effect of Artemin on LPS-induced nitric oxide synthesis in macrophages. METHOD: 1. Nitrite oxide (NO) production of RAW 264.7 cells was induced by LPS or LPS in combination with interferon-gamma (IFN gamma) in the presence or absence of Artemin. The amount of NO in the supernatant of RAW 264.7 cells was detected with Griess reagent. 2. Balb/c mice were injected with Artemin (i.m.) 50 mg.kg-1.d-1 x 3 d, and intraperitoneal macrophages were collected to detect the LPS-induced NO production. RESULT: LPS 1.0, 0.2 microgram.ml-1 or IFN-gamma 100u + LPS 1.0, 0.2, 0.04 microgram.ml-1 could induced a large amount of NO synthesis of RAW 264.7 cells. Artemin showed a significant inhibitory effect on LPS or IFN-gamma + LPS-induced NO production in a dose dependent manner. After treatment with Artemin, the response of Balb/c mice to LPS was reduced, which was showed by a decrease in NO production of intraperitoneal macrophages induced by LPS. CONCLUSION: Artemin could reduce LPS induced production of inflammatory factors resulting in the inhibition of inflammatory effects. PMID- 12776352 TI - [The protective effects of yi-xin-kang capsule on the structure and function of rat myocardial mitochondria]. AB - OBJECT: To observe the injury induced by Isoproterenol (ISO) on the structure and function of rat myocardial mitochondria and the protective effects of Yi-Xin-Kang Capsule (H303). METHOD: After injection of ISO, rat myocardial mitochondria were isolated. The activities of phospholipidase A2(PLA2) and Ca(2+)-ATPase, the content of phospholipids (PL) and high-energy phosphates, membrane lipid fluidity (LFU) and respiration function were determined respectively. RESULT: ISO induced severe injury to myocardial mitochondria. The activity of PLA2 was significantly activated. The intensity of LFU, the activity of Ca(2+)-ATPase, respiration function of mitochondria and the contents of PL and high-energy phosphates of myocardium tissue were significantly decreased. Oral administration of H303 significantly decreased the activity of PLA2. The contents of PL and LFU, the activity of Ca(2+)-ATPase were significantly increased. The respiration function of mitochondria was protected by accelerating III state respiration rate, improving ADP/O ratio and recovering the depressed respiration control rate. CONCLUSION: H303 possessed protective effect on structural and functional injury of mitochondria induced by ISO. PMID- 12776353 TI - [The protective effects of tea catechins on the injury of cerebral ischemia and reperfusion in rats]. AB - OBJECT: The protective effects of tea catechins on the injury of cerebral ischemia and reperfusion in rats were observed with rat cerebral ischemia and reperfusion model produced by bilaterally clamping the common carotid arteries and vagus nerves for 60 min and then removing the occlusion for 30 min. METHOD: Wistar rats were divided into normal control group, model group and catechin group (30 mg.kg-1 and 50 mg.kg-1 i.p., qd x 7 d). The changes of biochemical indications and pathology were observed. RESULT: The results presented that the concentration of MDA in brain tissue and serum of model rats were higher than that of control rats(P < 0.01). Compared with model group, tea catechin groups could decrease the concentration of MDA in brain tissue and serum, and tea catechin group (50 mg.kg-1 i.p., qd x 7 d) could improve the injury of blood brain barrier. CONCLUSION: Tea catechins have protective effects on the injury of cerebral ischemia and reperfusion in rats. PMID- 12776354 TI - [Cluster analysis of Dendrobium by RAPD and design of specific primer for Dendrobium candidum]. AB - OBJECTIVE: To analyze the genetic relationship of Dendrobium and design a specific primer to differentiate Dendrobium candidum effectively. METHOD: Random amplified polymorphic DNA (RAPD) technique was used to analyze the genetic relationships of 26 species of Dendrobium and one species of Ephemerantha. Dendrogram was constructed by UPGMA. According to the sequence of DNA fragment selected, Sangon 18 primer had been extended from 3' extreme to 20 bp in order to form a specific primer. RESULT AND CONCLUSION: This primer can be used to distinguish Dendrobium candidum from other Dendrobium effectively. Such a technique provides a new way for the identification of Chinese traditional medicines. PMID- 12776355 TI - [Determination of shikimic acid in fruit of Illiciaceae plants by HPLC with diode array detection]. AB - OBJECTIVE: To establish the determination method of Shikimic acid. METHOD: Using HPLC method as the determination method. The separation was performed in a SiO2 NH2 column with a mobile phase of Acetonitrile-2% H3PO4 water solution (95:5); The sample wavelength was 213 nm, reference wavelength 300 nm. RESULT: The average collection was 98.5%, RSD 1.67% (n = 5). CONCLUSION: This method is suitable for the determination of Shkimic acid in herb medicines and preparation containing shikimic acid. PMID- 12776356 TI - [Investigation on quality difference between Cordyceps of Sichuan and those of Xizang]. AB - OBJECT: The quality difference among the Cordyceps specimens which grew in various micro-ecological environment and between those from Kangding of sichuan and those from Naqu of Xizang was compared and analyzed. METHOD: The specimens from various spots in field were collected, their appearance properties were analyzed, content of adenosine was assayed, and their difference on ecological hereditary variation was discussed. RESULT: The biggest values of quality difference among the Cordyceps specimens from various micro-ecological environment can surpass that between specimen of Sichuan and specimen of Xizang. CONCLUSION: The effect of the micro-ecological environment on the quality of the Cordyceps might exceed that of extensive climates and region differences at times. PMID- 12776357 TI - [Studies on anamorph of Cordyceps sinensis (Berk) from Naqu Tibet]. AB - OBJECTIVE: To isolate and determine the strain of anamorph of Cordyceps sinensis collected from Naqu Tibet. METHOD: Isolating the strains of anamorph of Cordyceps sinensis collected from Baqing Shuoxian, Biru Counties by means of the ascope, tissue, ossified larva and body liquid. RESULT: Same strain of anamorph of Cordyceps sinensis was obtained with 4 different methods. CONCLUSION: The strain of Hirsutella sinensis is the anamorph of C. sinensis from Naqu Tibet. PMID- 12776358 TI - [Studies on Paecilomyces muscardine of Cordyceps sinensis host insect]. AB - OBJECTIVE: To Clarify the cause of the disease of Cordyceps sinensis host insect Paecilomyces muscardine. METHOD: Separately observing eggs, larvals, pupals, adults and their circumstances causing diseases in every stage. Adopting separately Hemolymph, bodies of dead insects and conidiospores; separating and culturing them, them observing and identifying the inoculum. Inoculating healthy larvals from the purely separated and cultured plants of inoculum. RESULT AND CONCLUSION: The original inoculum of the disease is Paecilongces frinoues, which may often be found in the period of larvals and seldom found in the period of pupals but never in that of adults and eggs. The main symptom of larval is dullness and weakness. But the typical Symptom is a layer of a kind of white power of conidiospore on the surface of the dead body which comes into being later stage. PMID- 12776359 TI - [Studies on the chemical constituents from Aquilegia ecalcarata Maxim]. AB - OBJECTIVE: To study the chemical constituents from Aquilegia ecalcarata. METHOD: Compounds were isolated with silica gel and polyamide chromatography and their structures were determined by spectral analysis and chemical evidence. RESULT: Five compounds were obtained and identified as beta-sitosterol, isoorientin-7-O glucoside, isovitexin-4'-O-glucoside, isovitexin-2'-O-rhamnoside, luteolin-7-O glucoside. CONCLUSION: All the compounds were isolated from this plant for the first time. PMID- 12776360 TI - [Isolation and elucidation of chemical constituents of shandanshaoyao decoction (II)]. AB - OBJECTIVE: To study the chemical constituents of Shandanshaoyao Decoction. METHOD: The sample was prepared by boiling water, then precipitated by 75% of ethanol. The ethanol soluble fraction was extracted by chloroform, ethyl acetate and n-butanol, respectively. The extracts were isolated using macroreticular resin, silica gel and polyamide column chromatography, and then the chemical constituents were identified by chemical and spectroscopic methods. RESULT: Nine compounds were isolated from the n-butanol extract and water soluble fraction, and elucidated as vitexin, quercetin, hyperoside, rutin, paeoniflorin, protocatechaldehyde, (+)-catechin, danshensu and citric acid respectively. CONCLUSION: The chemical constituents of traditional Chinese medicinal preparations Shandanshaoyao Decoction were reported for the first time. PMID- 12776361 TI - [Studies on chemical constituents of Stellera chamejasma L]. AB - OBJECTIVE: Isolate and identify the bioactive compounds from the root of Stellera chamejasma. METHOD: The compounds were extracted with solvents, isolated by column chromatography and identified by spectroscopic methods. RESULT: Seven compounds were isolated and identified as umbelliferone(1); daphnoretin (2); 2,6 dimethoxyl p-benzoquinone(3); (-)-eudesmin(4); (+)-matairesinol(5); lirioresinol B(6) and daucosterol(7). CONCLUSION: Compounds 3, 4 and 5 were isolated from the plant for the first time. PMID- 12776363 TI - [HPLC fingerprints identification of Panax ginseng C. A. Mey., P. quinquefolin L. and P. notoginseng (Burk. F. H. Chen)]. AB - OBJECTIVE: The method was established to identify Panax ginseng, P. quinquefolium and P. notoginseng. METHOD: Polaris C18-A analytical column (250 mm x 4.6 mm, 5 microns); acetonitrile-water as gradient eluent, flow rate 1.5 ml.min-1, detective wavelength at 203 nm. RESULT AND CONCLUSION: The fingerprints of P. ginseng, P. quinquefolium and P. notoginseng were obtained, and all ginsenosides were analyzed perfectly. The peak height ratio of ginsenoside Rg1 and ginsenoside Re was a suitable character to differentiate the three species from each other. PMID- 12776362 TI - [Studies on the chemical constituents from Myristica yunnanensis Y. H. Li]. AB - OBJECTIVE: To study the chemical constituents of Myristica yunnanensis. METHOD: Compounds were isolated using silica gel and reversed-phase C18 column chromatography, their structure were identified by means of spectral method. RESULT: Five known compounds were identified as otobain(I), beta-sitosterol(II), ergosterol(III), afzelin(IV) and quercetrin(V). CONCLUSION: Compounds IV and V were isolated first from Myristica. PMID- 12776364 TI - [Studies on protective effect of total flavonoids of Astragalus on liver damage induced by paracetamol]. AB - OBJECTIVE: To study the protective effect of total flavonoids of Astragalus (TFA) on the liver against large doses of paracetamol in mice. METHOD: After oral administration of TFA or Vitamin C 1 h prior to giving large dose of paracetamol in mice, the changes of paracetamol-induced mortality rate, serum enzyme level and liver damage degree were observed. RESULT: Paracetamol produced 80% mortality, within 24 hours of the administration of a dose of 1000 mg.kg-1 to the mice. Pre-treatment of the animals with TFA (100 mg.kg-1) or Vitamin C (1,000 mg.kg-1) reduced the death rate to 20% and 0% respectively. There was also a significant rise in the serum enzyme level of alanine transaminase (P < 0.001) and the area of liver necrosis (P < 0.001), 24 h after paracetamol (400 mg.kg-1) treatment. With pre-treatment with either TFA or Vitamin C, there was an obvious dose-dependent decrease in ALT levels and the area of hepatocellular necrosis. CONCLUSION: TFA has potential protecting effect against the paracetamol-induced hepatic damage. PMID- 12776365 TI - [Pattern recognition of stereoscopic features of the leaves epidermis of medicinal Curcuma plants in China by image analysis]. AB - OBJECTIVE: To provide new ways for classifying and identifying medicinal curcuma plants. METHOD: Based on classical taxonomy, the microscopic features of the epidermis of Curcuma plants in China were scored quantitatively by stereology and image analysis. RESULT: It showed that there were a lot of differences in density, size and shape of the epidermic cells among the medicinal Curcuma plants in China; that the average perimeter of the upper epidermic cells, average sectional area of the lower epidermis, stoma density and trichoma distribution of the upper and lower epidermis, etc. could be considered to be the main evidences for the microscopic identification of leaves of Curcuma. The 21 materials which belong to 11 species of Curcuma in China were systematized into 6 species, 1 species complex, 2 cultivated varieties. CONCLUSION: It can be concluded that stereology, and image analysis are advanced and feasible in pharmacognosy and taxonomy especially in the authentication of the relative and easily confused species. PMID- 12776367 TI - [Study on activity of inner inhibitory substances of Acanthopanax senticosus (Rupr. et Maxim) harms fruits and seeds]. AB - OBJECTIVE: By studying on the inner inhibitory substances of Acanthopanax senticosus fruits and seeds, the reason for dormancy of the Seeds was investigated. METHOD: First, the raw extracts of Acanthopanax senticosus fruits and seeds were prepared and determined by biological methods. Then, the ether extract of Acanthopanax senticosus fruits was prepared and the components were separated by paper chromatography and each section with different Rf value was determined, respectively. RESULT AND CONCLUSION: Inner inhibitory substances with high activity exists in Acanthopanax senticosus fruits and their outer coats, middle coats and the seeds. The inner inhibitory substances showed stronger inhibitory effect on the germination of Brassica chinensis seeds than the growth of its younger roots. The Rf 0.1 section of the ether extract of Acanthopanax senticosus fruit coats showed the strongest inhibitory activity on the germination of Brassica chinensis seeds and the growth of its young roots. The rough extracts of Acanthopanax senticosus fruits could inhibit the activity of the (alpha-amylase in the process of wheat seeds germination. The dormanvy of Acanthopanax senticosus seeds is related to the inhibitory substances with high activity in Acanthopanax senticosus fruits and seeds. PMID- 12776366 TI - [Studies on original plant of traditional Chinese drug "bai zhi" (radix Angelicae Dahuricae) and its closely related wild plants. I. Morphological and anatomical studies on "bai zhi" and closely related wild plants]. AB - OBJECTIVE: To supplement morphological and anatomical data for confirming the original plant of traditional Chinese drug "Bai Zhi" (Radix Angelicae Dahuricae). METHOD: Morphologocal observation and anatomical study were made on 4 cultivated breeds and closely related wild plants of "Bai Zhi". RESULT: According to morphological and anatomical characteristics discovered in this paper, 7 samples noted above could be divided into 3 groups: 1. 4 breeds ("Chuan Bai Zhi", "Hang Bai Zhi", "Qi Bai Zhi" and "Yu Bai Zhi") and Angelica dahurica var. formosana; 2. A. dahurica; 3. A. porphyrocaulis. CONCLUSION: In the morphological and anatomical point of view, A. dahurica var. formosana is closer to traditional Chinese drug "Bai Zhi" than others. PMID- 12776368 TI - [Studies on the chemical constituents from Chrysanthemum morifolium Ramat]. AB - OBJECTIVE: To study the chemical constituents of Chrysanthemum morifolium. METHOD: Separated the constituents by means of chromatography and identified their structures by chemical and spectroscopic analysis. RESULT: Three compounds of flavonoid were identified as acacetin-7-O-beta-D-glucoside, apigenin-7-O-beta D-glucoside and luteolin-7-O-beta-D-glucoside. CONCLUSION: These compounds were obtained from C. morifolium for the first time. PMID- 12776369 TI - [Chemical constituents from the fruit of Rosa bella Rehd. et Wils]. AB - OBJECTIVE: To investigate the chemical constituents from the fruit of Rosa bella and its bioactivity. METHOD: The compounds were isolated from the methanol extract by column chromatography, then identified by 1H, 13C NMR and by comparison with authentic samples. The unorganized hoptoad heart was used for testing the contracting force of cardiac muscle and enzymology for measuring the bloody lipid of serum in rats. RESULT: Seven compounds were isolated and identified as pomolic acid, tiliroside, euscaphic acid, daucosterol, quercetin, oleanolic acid and beta-sitosterol, respectively. In the concentration of 0.4-1.0 mg.ml-1, the methanol extract had obvious cardiotonic effect. While at 400 mg.kg 1, there are no obvious effect to the TC, HDL-ac and TG of serum in rats. CONCLUSION: The isolated compounds may be the active components of this plant. PMID- 12776370 TI - [Non-anthraquinone constituents from Rheum sublanceolatum C. Y. Cheng et Kao]. AB - OBJECTIVE: Study on the non-anthraquinone constituents from rhizoma and radix of Rheum sublanceolatum. METHOD: The constituents were isolated through column chromatography and identified on the basis of their physiochemical and spectral data. RESULT: Six non-anthraquinone constituents were isolated and identified as n-octacosanic acid, sitosterol, daucosterol, 2-methyl-5-carboxymethyl-7 hydroxychromone, piceatannol and 6-hydroxymusizin-8-O-beta-D-glucopyranoside. CONCLUSION: All these compounds were firstly isolated from R. sublanceolatum. PMID- 12776371 TI - [Studies on chemical constituents of oligostilbenes from Vitis davidii Foex]. AB - OBJECTIVE: To study the chemical constituents of oligostilbenes from Vitis davidii. METHOD: Compounds were isolated with polystyrene resin RA, silica gel and C18 column chromatography. The structures were elucidated by means of spectroscopic evidence. RESULT: Seven compounds were obtained and identified as resveratrol, heyneanol A, ampelopsin E, amurensin B, (+)-epsilon-viniferin, vitisin A and amurensin G. CONCLUSION: All the compounds were isolated from this plant for the first time. PMID- 12776372 TI - [Effect of Rg2 on hemodynamics of hemorrhagic shock and its antioxidation in dogs]. AB - OBJECTIVE: To investigate the protective effect of Rg2 on hemodynamics of hemorrhagic shock and its antioxidant properties. METHOD: Twenty mongrel dogs were randomly divided into Rg2 group, Shen Mai group and control group. The hemorrhagic shock model was built in all dogs by artery bleeding and mean arterial blood pressure was kept < 5.33 kPa for 4.5 hours. Rg2 0.5-1.0 mg.kg-1 and Shen Mai 100 mg.kg-1 were intravenously administered after hemorrhagic shock appeared. RESULT: Rg2 significantly enhanced blood pressure, LVSP and +/- dp/dtmax on hemorrhagic shock dogs who had lost compensation ability. Rg2 0.5-1.0 mg.kg-1 could reduce serum MDA, increase superoxide dismutase activity and prolong survival rate of dogs after shock. All these data have statistically significant when compared to control group (P < 0.05, P < 0.01, P < 0.001). CONCLUSION: These results suggest that Rg2 is more potent and effective than Shen Mai in improving hemodynamic state and activiting SOD on hemorrhagic shock dogs. PMID- 12776373 TI - [The analgesic and antispasmodic effects of guang tong xiao aerosol of TCM]. AB - OBJECTIVE: To observe the analgesic and antispasmodic effects of Guang Tong Xiao Aerosol (GTXA). METHOD: Writhing test and tail-flick of physical stimulation were made to study the analgesic effect on mice and rats. RESULT AND CONCLUSION: GTXA given by gastrogavage in dose of 18.75 g.kg-1 or 12.50 g.kg-1 could markedly raise the pain threshold after chemical stimulation in mice and physical stimulation in rats, and had antispasmodic effects. PMID- 12776374 TI - A two pronged test for surgical decision-making in the cognitively impaired patient. PMID- 12776375 TI - Ethical issues in the use of covert video surveillance in the diagnosis of Munchausen syndrome by proxy: the Atlanta study--an ethical challenge for medicine. PMID- 12776376 TI - Informed consent in clinical practice. PMID- 12776377 TI - Dialogue as a means to resolve ethical issues in health care. PMID- 12776378 TI - Genetic information and research: emerging legal issues. PMID- 12776379 TI - Roman Catholic norms and the allocation of critical care resources. PMID- 12776380 TI - Psychosocial correlates of incidence of attacks in children with Familial Mediterranean Fever. AB - This study tested the relationship between psychosocial factors and incidence of Familial Mediterranean Fever (FMF) attacks. Forty-five children with FMF were studied retrospectively. Parents assessed their child's hostility, perceived control, illness-behavior encouragement (IBE), family dysfunction, and reported number of attacks during the last 12 months. Hostility was positively correlated with number of attacks, especially in children below age 10 and in girls. Family dysfunction was positively correlated with attacks in girls and in children at or above age 10. IBE was inversely correlated with attacks in older children. In children below age 10, number of siblings was positively correlated with attacks, and negatively correlated with attacks in the older group. Psychosocial factors explained 27% of the variability in attacks, after controlling for age and number of siblings, with hostility remaining the only significant predictor of attacks. These findings, if replicated in a prospective study, may guide interventions for preventing FMF attacks. PMID- 12776381 TI - Prospective association between distress and mammography utilization among women with a family history of breast cancer. AB - In this study we sought to evaluate the prospective association between psychological distress and mammography utilization among women with a family history of breast cancer. We evaluated the association of cancer worry, cancer specific distress, and general distress with mammography utilization after controlling for potential confounders. The results revealed that 74% of our sample had obtained a mammogram within 12 months of the baseline assessment. Logistic regression models revealed that after controlling for potential confounding variables, cancer worry and general distress were independent predictors of mammography utilization. Specifically, women who reported higher levels of worry and/or distress at baseline were less likely to report having received a mammogram in the 12 months following the baseline assessment. These results are in contrast to the only other prospective study in this population. Additional research is needed to determine the prospective association between distress and adherence and to identify potential mechanisms for such an association. PMID- 12776382 TI - Age differences in symptoms of depression and anxiety: examining behavioral medicine outpatients. AB - This study examined whether symptoms of depression and concomitant anxiety differed between older and younger medical outpatients referred to a behavioral medicine clinic. In a sample of 178 male veterans aged 21-83 years, older adults (> or = 60 years) reported lower overall depressive symptoms on the Beck Depression Inventory (BDI) and anxiety symptoms on the State-Trait Anxiety Inventory than did younger adults ( < 60 years). Depressive symptoms were highly prevalent. Among older adults, 60.0% scored 10 or higher on BDI and 33.8% scored 16 or higher. Among younger adults, 70.8% scored 10 or higher on BDI, and 48.7% scored 16 or higher. The age difference in overall depressive symptoms was driven by cognitive-affective symptoms. While older adults had lower cognitive-affective symptoms than did younger adults, the two groups did not differ on somatic performance symptoms. these results suggest the importance of assessing cognitive affective depressive symptoms in both older and younger male medical outpatients. PMID- 12776383 TI - Illness representation and change in dietary habits in hypercholesterolemic patients. AB - The association between a patient's illness representation and long-term changes in treatment adherence still needs to be clarified. AIM: to evaluate the association between the representation of hypercholesterolemia and dietary modifications over 1-year, controlling for biopsychosocial factors. METHODS: 208 hypercholesterolemic Caucasian patients. Measures of dietary intake, illness representation, and biopsychosocial factors were collected at baseline, 3, 6, and 12 months. RESULTS: Two multiple regressions were performed on the 1-year change in dietary habits; one with subjects with elevated fat consumption at baseline, another with individuals with low-moderate fat consumption. Higher scores on items reflecting an accurate illness representation at baseline were associated with greater dietary modification among patients of the first group. For patients with low-moderate total fat consumption, maintenance of dietary habits was associated to lower scores on the perception of stress and symptoms. CONCLUSION: To optimize adherence, intervention on illness representation should be adjusted according to initial dietary habits. PMID- 12776384 TI - Sources of anticipatory distress among breast surgery patients. AB - Surgical consultation concerning the possibility of breast cancer is a distressing experience, and having to take the next step of breast surgery even more so for many women. However, the sources of variability in such presurgical distress are not well understood. Sixty-one women (mean age = 51) were recruited immediately following surgical consultation in which a recommendation of breast surgery (excisional biopsy/lumpectomy) was made. Patients completed measures of distress, worry about cancer and surgery, trait anxiety, optimism and pessimism prior to surgery. Surprisingly, results revealed no effect of surgeon-provided information concerning preliminary diagnosis on patient distress. Rather, worry about what the surgeon might find concerning the breast mass during surgery, worry about having to go through the operative procedures, and patient optimism were the only factors that uniquely contributed to patient distress (p's < 0.05). This study provides a foundation for future clinical interventions to reduce presurgery distress. PMID- 12776386 TI - The implications of core competencies for psychiatric education and practice in the US. AB - Physician competence is a universal concern, one that Canada and the US have addressed in differing, but also in similar, ways. Focusing on the roles physicians play, the Royal College of Physicians and Surgeons of Canada (RCPSC) has implemented a uniform procedure for developing and assessing competencies. The US does not have a parallel body but has instead different organizations responsible for different phases of medical education from residency through practice. These groups are working with 6 categories of core competencies to be used for assessment purposes. The categories are patient care, medical knowledge, interpersonal and communication skills, practice-based learning and improvement, professionalism, and systems-based practice. This article presents the US core competencies for psychiatric practice as they are currently being implemented through the American Board of Psychiatry and Neurology, Inc. PMID- 12776387 TI - Mastering CanMEDS roles in psychiatric residency: a resident's perspective. AB - Postgraduate trainees in psychiatry are being evaluated on their proficiency at competencies that comprise the physician roles identified by the CanMEDS 2000 Project. This paper provides an overview of each CanMEDS role and its associated competencies and suggests strategies to help residents prepare for the new format of the Royal College of Physicians and Surgeons (RCPSC) certification examination in psychiatry. PMID- 12776385 TI - AIDS-related grief and coping with loss among HIV-positive men and women. AB - AIDS-related grief was examined and its association with coping among HIV positive men and women explored. AIDS-related grief, psychological distress and coping were examined among a sample of 268 HIV-infected individuals, diverse with respect to gender, ethnicity, and sexual orientation. Participants exhibited elevated scores on measures of grief reaction and psychological distress including depressive symptoms, anxiety, and traumatic stress related to their losses. Hierarchical regression analyses revealed that severity of grief reaction was associated with escape-avoidance and self-controlling coping strategies, type of loss, depressive symptoms, and history of injection drug use. Interventions are needed to enhance coping and reduce psychological distress associated with the unique bereavement experienced by people living with HIV- and AIDS-related grief. PMID- 12776388 TI - Residency training: challenges and opportunities in preparing trainees for the 21st century. AB - The future will see increased medicalization of psychiatry and will demand changes in training that better prepare residents for the realities of practice in a sustained period of physician shortage. Residency programs will need to move from the current apprenticeship model of training to competency-based programs built on the CanMEDS 2000 articulation of physician roles. Training will need to focus on evidence-based treatments, more efficient models of health care delivery, more attentive tracking of resident clinical work, and more reliable and standardized methods of evaluating resident competencies. PMID- 12776389 TI - Patient characteristics associated with nonprescription drug use in intentional overdose. AB - OBJECTIVE: Over-the-counter (OTC) medications remain freely available to suicidal patients, despite their potential lethality and common use in suicide. The study's main objective was to identify patient characteristics, particularly psychiatric diagnosis associated with the use of OTC medications in intentional overdose. METHODS: We retrospectively reviewed 95 charts from patients who presented to St Paul's Hospital from August 1, 1997, to July 31, 1998, with a discharge diagnosis of intentional drug overdose. Univariate analysis was carried out to identify potential risk markers for OTC medication use, and logistic regression was performed using these variables. RESULTS: When the variables age, sex, and concurrent psychiatric diagnoses were controlled, use of OTC medications in overdose was significantly lower in patients with a DSM-IV diagnosis of substance abuse (OR 0.11, P = 0.005) and in those who possessed prescription medications at the time of overdose (OR 0.18, P = 0.007). Most patients in this cohort (82%) had at least 1 of these 2 traits. Although not statistically significant, younger patients appeared more likely to choose OTC medications for overdose. CONCLUSION: Suicide-prone patients with a diagnosis of substance abuse and who possess prescription medications are unlikely to use OTC medications in overdose. For this cohort, this represents a relatively small proportion of patients whom clinicians should consider to be at greater risk for attempting suicide when using OTC medication, especially acetaminophen. PMID- 12776390 TI - The Canadian Psychiatric Association practice profile survey: I. Methods and general sample characteristics. AB - OBJECTIVE: To describe the rationale, methodology, and general sample characteristics of the Canadian Psychiatric Association (CPA) practice profile survey, a national survey of psychiatrists and psychiatric practice. METHOD: Mail in interviews were sent to all Canadian psychiatrists listed in their provincial registers and to all active CPA members (total = 3628). Respondents provided general information about their professional activities for one 24-hour day and detailed information for 1 randomly selected hour. Patient information--including sociodemographics, diagnostic profiles, functioning levels, risk of harm to self or others, and disposition--was elicited for 1 patient seen during the random hour as well as for the most seriously ill patient receiving clinical services that day. RESULTS: There was a 45.5% response rate. Questionnaires completed by nonpsychiatrists or with a large percentage of missing or incorrect data were eliminated (107 surveys), resulting in a final sample size of 1570. CPA members and those from Western Canada responded at a higher rate to the survey. The results suggest some cause for concern about future manpower shortages. Most psychiatrists practise eclectically, seeing patients across the life-span, and working in both community and institutional settings. The old and the young appear to be underserviced, compared with adults. CONCLUSIONS: This study represents an important step forward in evaluating the profile and activities of the profession. PMID- 12776391 TI - The Canadian Psychiatric Association practice profile survey: II. General description of results. AB - OBJECTIVE: To provide an overview of the results of the Canadian Psychiatric Association (CPA) practice profile survey (PPS), a national survey of psychiatrists and psychiatric practice. METHOD: Mail-in interviews were sent to all Canadian psychiatrists listed in their provincial registers and to all active CPA members (total = 3628). Respondents provided general information about their professional activities for one 24-hour day and detailed information for 1 randomly selected hour. Patient information--including sociodemographics, diagnostic profiles, functioning levels, risk of harm to self or others, and disposition--was elicited for 1 patient seen during the random hour as well as for the most seriously ill patient receiving clinical services that day. RESULTS: Psychiatrists work 10 hours daily on average and take calls for 5 hours. Sixty percent of the overall work time is in the provision of direct patient care, and fee-for-service payments account for 55% of hours worked. Forty percent of the clinical work is provided in a hospital setting, and 34% is in a private office. Agency work accounted for only 6% of clinical hours worked. Relatively few practitioners provide services to children, older, or forensic patients. The average patient seen is female, aged 40 years, unmarried or with a marital disruption, significantly impaired in multiple areas of functioning, and likely to suffer from depression (21%), schizophrenia (14%), an anxiety disorder (13%), or bipolar disorder (12%). Comorbid Axis I and Axis II disorders are common (each over 30%) and fairly high rates of suicidal (15% to 30%) and homicidal (10% to 20%) risk are present. CONCLUSIONS: This paper suggests a wide diversity of practice in psychiatry in Canada, with services being provided to a wide range of individuals with many different conditions. PMID- 12776392 TI - Effect of depression on stroke morbidity and mortality. AB - OBJECTIVES: This narrative review examines the evidence and discusses the clinical relevance of depression as a risk factor for stroke morbidity and mortality. It also proposes recommendations for future research. METHODS: We used the Medline computer database to search the relevant original studies published in English from January 1966 to December 2001. Our key words were as follows: depressive disorder, cerebrovascular disease, stroke, vascular risk factors, and mortality. Articles that investigated the relation between antecedent depression and subsequent stroke morbidity and mortality were collected and reviewed. RESULTS: Since 1990, 8 prospective studies have been published. Among these 8 studies, 6 addressed depression and stroke morbidity, 1 investigated the association of depression with stroke morbidity and stroke mortality, and 1 investigated the association with stroke mortality only. Of 7 studies examining the independent effect of depression on stroke morbidity, 6 were positive. With regard to stroke mortality, 2 studies found an independent association between depression and specific stroke mortality. The contributions and methodological limitations of these studies are discussed. CONCLUSIONS: Emerging data suggest an association between depressive symptoms and increased risk for stroke morbidity and mortality. More methodologically sound studies are needed to elucidate causal pathways that link depression and cerebrovascular disease. They are also needed to determine the effect of depression intervention on reducing the risk of cerebrovascular events. Information on author affiliations appears at the end of the article. PMID- 12776393 TI - Switch to mania upon discontinuation of antidepressants in patients with mood disorders: a review of the literature. AB - OBJECTIVE: To review the literature for reported cases of mania related to discontinuing antidepressant treatment, as well as for possible explanations of this phenomenon, and to present a case report. METHOD: We undertook a literature review through the PubMed index, using the key words mania, antidepressant withdrawal, and antidepressants in bipolar disorder. We reviewed 11 articles featuring 23 cases. Where available, we noted and tabulated certain parameters for both bipolar disorder (BD) and unipolar depression. We use a case example to illustrate the phenomenon of mania induced by antidepressant withdrawal. RESULTS: For patients with unipolar depression, we found 17 reported cases of mania induced by antidepressant withdrawal. Antidepressants implicated included tricyclic antidepressants (TCAs) (12/17), monoamine oxidase inhibitors (MAOIs) (2/17), trazodone (1/17), mirtazapine (1/17), and paroxetine (1/17). For patients with BD, we found 19 reported cases of mania induced by antidepressant withdrawal, including our own case example. Of these, selective serotonin reuptake inhibitors (SSRIs) (10/19), TCAs (4/19), MAOIs (2/19), and serotonin norepinephrine reuptake inhibitors (SNRIs) (2/19) were implicated. CONCLUSION: Our case report supports the observation of antidepressant withdrawal-induced mania in patients with BD. It is distinguishable from antidepressant-induced mania, physiological drug withdrawal, and mania as a natural course of the illness. Many theories have been put forward to explain this occurrence. Noradrenergic hyperactivity and "withdrawal-induced cholinergic overdrive and the cholinergic-monoaminergic system" are the 2 most investigated and supported models. The former is limited by poor clinical correlation and the latter by its applicability only to anticholinergic drugs. PMID- 12776394 TI - Acute neuroendocrine response to sexual stimulation in sexual offenders. AB - BACKGROUND: Several pharmacotherapeutic approaches have confirmed the influence of neuroendocrine parameters on sexual desire, function, and fantasies in men; however, the relevance of acute neuroendocrine changes in mediating heightened sexual drive remains unknown. We recently demonstrated that plasma prolactin substantially increases following orgasm in healthy men, suggesting a feedback mechanism for peripheral prolactin in the control of acute sexual arousal. Because prolactin appears to play a regulatory role in acute sexual drive, we initiated this study to see whether sexual offenders with a high sexual drive have a different neuroendocrine response to sexual arousal. This study compares the prolactin response to orgasm of sexual offenders with high sexual drive and that of healthy subjects with average sexual drive. METHODS: From a subject pool of 150 inpatients held because of sexual crimes, we recruited 10 volunteers, based on their high sexual drive according to an intensive, semistructured clinical interview. We defined sexual drive by a short refractory period and strong sexualization, or a high frequency of sexual stimulation. We analyzed the acute psychoneuroendocrine response to sexual arousal and orgasm continuously before, during, and after masturbation-induced orgasm in patients and control subjects. RESULTS: Sexual offenders demonstrated higher sexual desire (P < 0.001) and function (P < 0.001) and a more positively perceived refractory period (P < 0.05). Both groups displayed a prolonged, significant increase in prolactin plasma levels after orgasm (P < 0.001). Sexual offenders did not differ from control subjects in neuroendocrine response to sexual arousal and orgasm. CONCLUSIONS: These data demonstrate that sexual offenders with a high sexual drive do not differ from control subjects in the postorgasmic neuroendocrine response, particularly in prolactin release. This study confirms that factors other than peripheral hormones influence deviant sexual behaviour. PMID- 12776396 TI - Influence of season and latitude in a community sample of subjects with bipolar disorder. AB - OBJECTIVE: To report on the prevalence of seasonal bipolar disorder (BD) and the impact of latitude in a community sample in the province of Ontario. METHOD: This study used the telephone-administered Depression and Seasonality Interview. Exact latitude was determined for each participant. RESULTS: Overall, 14 of 62 (22.6%) subjects with BD had the seasonal subtype of BD. Latitude did not appear to influence the proportion of subjects with the seasonal subtype. CONCLUSIONS: We identified a seasonal pattern of illness in a proportion of subjects with BD. PMID- 12776395 TI - Weight gain in first-episode psychosis. AB - OBJECTIVE: To examine the extent of weight gain in the first year of treatment in an early psychosis program. METHOD: Subjects were 114 individuals who had experienced a first episode of psychosis and had completed 1 year in a comprehensive first-episode program. Weight and body mass index were calculated on entry to the program and at 6 and 12 months. Most of the subjects were all being prescribed second-generation antipsychotics. RESULTS: Significant increases in mean weight were observed in these young individuals over the course of the first year of treatment. CONCLUSIONS: If we are to work toward optimum treatment for first-episode subjects then potential weight gain needs to be addressed at the beginning of treatment and monitored during treatment. PMID- 12776397 TI - Biological factors and adolescent alcohol use. PMID- 12776398 TI - Minor strokes related to paroxetine discontinuation in an elderly subject: emergent adverse events. PMID- 12776400 TI - Behaviour therapy for dizziness? PMID- 12776399 TI - Quetiapine reduces flashbacks in chronic posttraumatic stress disorder. PMID- 12776401 TI - Involuntary treatment of a patient with factitious disorder: a paradox? PMID- 12776402 TI - Expanding the role of nurses. PMID- 12776403 TI - An update on the E/M codes and documentation guidelines. PMID- 12776404 TI - Seven ways to help your hospital stay in business. PMID- 12776405 TI - Implementing an EMR system: one clinic's experience. PMID- 12776406 TI - Dealing with a health system that fails to honor your employment agreement. PMID- 12776407 TI - Caring for frequent-visit patients. PMID- 12776408 TI - Improving patient care. Group visits 101. PMID- 12776410 TI - The nature of nursing homes. PMID- 12776411 TI - HIPAA takes effect. PMID- 12776412 TI - Bugged. AB - This was a great save. The crew could easily have missed the presentation of anaphylaxis and let the window for treatment with epinephrine slip away. This patient was in anaphylactic shock. There were no signs that supported a traumatic injury, and that, combined with diaphoresis, urticaria and tachycardic central pulse, contributed to the suspicion of anaphylaxis. Anaphylaxis is classified as distributive shock. This type of shock is caused by profound systemic vasodilation, and the heart is unable to increase output enough to maintain blood pressure. Other causes of distributive shock include sepsis and spinal cord injury. It is rare to have both hypotension and wheezing in such cases. In an anaphylactic reaction, an allergen, such as a food protein, medication, insect venom or latex, is introduced into the body. The mast cells of the immune system have a protein on their surface called IgE antibodies (Immunoglobulin E). The mast cells are filled with histamines [table: see text] and leukotrienes, which are chemical mediators. These are released when the allergen reacts with the IgE antibodies. When these mediators are released, they cause smooth-muscle constriction in the respiratory and gastrointestinal tracts, resulting in wheezing, stridor, nausea, vomiting and diarrhea. They also cause vascular dilation, leading to edema and urticaria. Most patients will present with either profound vascular effect (shock) or wheezing; this is a rather rare presentation of a patient having both. The medication best suited to counteract the effects of these medicators is epinephrine. Epinephrine is an alpha- and beta-agonist, acting to constrict the vasculature and dilate the smooth muscles in the bronchial tree. Antihistamines can alleviate symptoms of anaphylaxis, but should only be used in addition to epinephrine, not as a substitute. In life-threatening reactions, epinephrine must be given quickly and in a form that the body can distribute. Use of the subcutaneous route with a solution mixed at 1:1,000 dilution is appropriate in most patients, but if the patient is in profound shock and not perfusing the skin (pale, cold, clammy skin), then a more diluted concentration must be given i.v. at a slow rate (1 cc every minute of the 1:1,000 dilution) until the patient recovers. If i.v. access is delayed or not available, give the 1:1,000 dilution intramuscularly, in the tongue or down the endotracheal tube. Refer to your local protocols for dosage, but the usual dose of epinephrine is 0.3-0.5 mg, or 0.01 mg/kg in a child. There are more than 40 million people in the U.S. with allergic histories that place them at risk for developing anaphylaxis. Each year over 5,000 deaths are attributed to anaphylaxis. The risk of death from anaphylaxis increases with a more rapid onset of signs and symptoms. Up to 25% of patients will experience a biphasic reaction. This means there is a recurrence of symptoms several hours after the initial reaction, and it is prudent to observe patients for a period of time following their initial treatment. PMID- 12776413 TI - "Know thy ambulance". PMID- 12776414 TI - Detecting mechanism of injury. PMID- 12776416 TI - Hip fractures and dislocations revisited. PMID- 12776415 TI - Pelvic trauma. PMID- 12776417 TI - EMS mythology, Part 3. EMS myth #3: Critical incident stress management (CISM) is effective in managing EMS-related stress. PMID- 12776418 TI - The universal truths of writing incident reports. PMID- 12776419 TI - Scene safety vs. duty to care. PMID- 12776420 TI - EMS in Canada: Part 1. PMID- 12776421 TI - CDC's hand washing guidelines. PMID- 12776422 TI - How to satisfy preferences of rehab residents in a nonselect menu facility. PMID- 12776423 TI - [Studies on original plant of traditional Chinese drug "bai zhi" (radix Angelicae Dahuricae) and its closely related wild plants. II. Karyological and pollen morphological studies on "bai zhi" and closely related wild plants]. AB - OBJECTIVE: To supplement cytobiological and pollen morphological data for confirming the original plant of traditional Chinese drug "Bai Zhi". METHOD: Karyological study and pollen observation were made on "Bai Zhi" and its closely related wild plants. RESULT: Similarities and differences of "Bai Zhi" and its closely related wild plants were found. CONCLUSION: 1. 4 cultivated breeds of "Bai Zhi", Angelica dahurica, A. dahurica var. formosana, A. porphyrocaulis are really closely related plants. 2. A. dahurica var. formosana is closer to traditional Chinese drug "Bai Zhi" than others. PMID- 12776424 TI - [A study on quality standard of Sambucus chinensis Lindl]. AB - OBJECTIVE: To offer evidences for exploiting the plant resources and drafting the quality standards. METHOD: Studies of pharmacognosy. RESULT: The medicinal properties and the microscopic characteristics of the stem, leaves, rhizome and powder were described, drawn and physicochemically identified. CONCLUSION: A convenient and effective method for identifying the herb has been established. PMID- 12776425 TI - [Comparative studies on the fatty acids contained in four species of medicinal plants from family Euphorbiaceae and their endophytic fungi]. AB - OBJECTIVE: The relation of four species of medicinal plants from family Euphorbiaceae and their endophytic fungi was studied to find the source of active substances for developing new pharmaceutical resources. METHOD: The main fatty acids contained in Sapium sebiferum, Euphorbia pekinensis, Euphorbia helioscopia, Bischofia polycarpam and their 28 strains of endophytic fungi were compared and analysed by GC. RESULT: The main fatty acids of the plants are: alpha-linolenic acid, palmitic acid, linolenic acid and oleic acid. Linolenic acid, palmitic acid and oleic acid are the main fatty acids of the endophytic fungi. CONCLUSION: The fatty acids could be produced by the endophytic fungi, which could be used as a factor for identification. There are great differences at the contents of alpha linolenic acid between the plants and their endophytic fungi, which were suggested to be related with the nutrition absorption and the relationship between the endophytes and the host plant. PMID- 12776426 TI - [Chemical constituents from Salix oritrepha Schneid]. AB - OBJECTIVE: To study chemical constituents of the above ground part of Salix oritrepha. METHOD: Using chromatography to isolate compounds and their structure were identified by physical, chemical and spectral techniques. RESULT: Four compounds were isolated and elucidated as beta-sitosterol, beta-sitosteryl-3-O glucoside, friedelin, luteolin-7-O-glucoside. CONCLUSION: All of them were isolated from S. oritrepha for the first time. Friedelin from the plants of genus Salix for the first time. PMID- 12776427 TI - [Studies on chemical constituents of Umbilicaria esculenta (Miyoshi) minks]. AB - OBJECTIVE: To study the chemical constituents from the fruit bodies of Umbilicaria esculenta. METHOD: Five compounds were isolated from the ethanolic extract of this plant by all kinds of column chormatography, and structures were identified by spectroscopic analysis. RESULT: Five compounds were identified as ethyl orsellinate, orsellinic acid, orcinol, lecanoric acid and lecanorin. CONCLUSION: Ethyl orsellinate, orsellinic acid, orcinol and lecanorin were isolated from this plant for the first time. PMID- 12776428 TI - [Studies on chemical constituents of Fragaria ananassa Duch]. AB - OBJECTIVE: To study the chemical constituents from the fruit of Fragaria ananassa. METHOD: Using chromatographic methods to isolate compounds and chemical and spectral methods to elucidate their structures. RESULT: Three compounds, 9, 19-cyclolanost-24-en-3-ol(1), 14-methyl-stigmasta-7, 24(28)-dien-3-ol(2) and beta sitosterol(3) were isolated from the freeze-dried powder. CONCLUSION: All of the compounds were obtained from this plant for the first time. PMID- 12776430 TI - [HPLC determination of oleanolic acid and llrolic acid in Chinese medicinal herbs]. AB - OBJECTIVE: To develop a new method for simultaneous determination of oleanolic acid and llrolic acid in Chinese medicinal herbs at the same time. METHOD: HPLC was carried out on a Shim-pack CLC-ODS column using MeOH-H2O-HOAc-TEA (83:17:0.04:0.02). RESULT AND CONCLUSION: The average recovery of oleanolic acid and llrolic acid was 103.3 +/- 2.07% and 102.7% +/- 0.65% respectively. PMID- 12776429 TI - [Studies on chemical constituents from Anemone anhuiensis Y. K. Yang N. Wang et W. C. Ye]. AB - OBJECTIVE: To investigate the constituents from the roots of Anemone anhuiensis. METHOD: To isolate chemical constituents, using solvent extraction together with column chromatography, FAB-MS and NMR methods were employed for strutural elucidation. RESULT AND CONCLUSION: Three compounds were isolated and elucidated as oleanolic acid 28-O-alpha-L-rhamnopyranosyl-(1-->4)- beta-D-glucopyranosyl-(1- >6)-beta-D-glucopyransyl ester (cussonoside B), 3-O-alpha-L-rhamnopyranosyl-(1- >2)-beta-D- xylopyranosyl oleanolic acid 28-O-alpha-L-rhamnopyranosyl-(1-->4) beta-D- glucopyranosyl-(1-->6)-beta-D-glucopyransyl ester (flaccidoside II) and 3 O-beta-D-glucopyranosyl-(1-->2)-beta-D-xylopyranosyl oleanolic acid 28-O-alpha-L rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl- (1-->6)-beta-D-glucopyransyl ester (flaccidoside III), respectively. All of them were isolated from the plant for the first time. PMID- 12776431 TI - [Protection against paracetamol-induced hepatic damage using total flavonoids of Astragalus]. AB - OBJECTIVE: To study the mechanism of the protection by total flavonoids of Astragalus protection against paracetamol-induced hepatic damage. METHOD: Analysing paracetamol and its metabolites in mice urine by HPLC and studying the mechanism of anti-damage induced by paracetamol using experiment module of pentobarbital-induced sleeping time. RESULT: Administration of large doses of paracetamol to C57BL/6J mice produced significant hepatic injury with marked elevation in serum ALT activity and severe hepatocellular necrosis. TFA showed a good protective capability against paracetamol-induced hepatic injury. TFA had no marked effect on paracetamol and its metabolites except for the mercapturate conjugate. The concentration of mercapturate change decreased with increasing TFA dose. TFA had no effect on the pentobarbital metabolites (P > 0.05). However, paracetamol (400 mg.kg-1) prolonged the sleeping time (by 110 min relative to the controls, P < 0.001). The TFA (P < 0.005) caused significant reduction in paracetamol-prolonged pentobarbital-induced sleep. CONCLUSIONS: The mechanism of TFA's protective effect against the paracetamol-induced damage may be related to the inhibition of some metabolism progress of paracetamol and the reduction of the toxicity metabolite such as mercapturate-conjugate. PMID- 12776432 TI - [Effects of fermented Cordyceps powder on pulmonary function in sensitized guinea pigs and airway inflammation in sensitized rats]. AB - OBJECTIVE: To study Cordyceps (artificial fermented Cordyceps sinensis(Berk.) Sacc) powderin the treatment of asthma in the animal models. METHOD: Pulmonary function and airway inflammation in vivo were investigated. RESULT: Cordyceps, 5g.kg-1(ig), significantly inhibited bronchial challenge of ovalbumin-induced change of RL and Cdyn (P < 0.05) and inhibited antigen-induced increase of eosinophils in the BALF of rats (P < 0.05). CONCLUSION: The results suggested cordyceps could be applied for the prevention and cure of asthma. PMID- 12776433 TI - [Study of the antiosteoporosis effects of bushenqianggu capsule]. AB - OBJECTIVE: To study the antiosteoporosis effects of Bushenqianggu capsule. METHOD: Observing the pharmacological effects of Bushenqianggu capsule with blank and positive control, and then analyzing the data with biological statistics. RESULT: The testis and uterus weight index of "the kidney assessed" model mice, weight of thighbone and blood concentration of Ca2+ increased significantly. Meanwhile, the thighbone mass density of castrated rats was improved and the incidence of the thighbone fracture in the osteoporosis model rats induced by retinoic acid was reduced. Besides, it had analgesia and anti-inflammatory effects. CONCLUSION: Bushenqianggu capsule has a good effect on osteoporosis, which provides evidence for clinical use. PMID- 12776435 TI - Dracunculiasis eradication. PMID- 12776434 TI - Travel advice for Toronto (Canada) lifted. PMID- 12776436 TI - Models of masculinity: sex education, the United States Public Health Service, and the YMCA, 1919-1924. AB - In 1918, the U.S. Public Health Service (PHS) told American parents that "it is no longer possible for you to choose whether your child will learn about sex or not." According to the PHS, most American boys learned about sex from "improper sources" by the age of nine. The "unfortunate effect of these early impressions" had, PHS warned, not only resulted in a gross misunderstanding of sex, but also been a major factor in the spread of venereal disease (The Parents' Part [the U.S. Public Health Service, 1918], p. 5). To counter and correct this miseducation, PHS joined with the Young Men's Christian Association (YMCA) to create a sex education program aimed at adolescent boys. Officially launched in the spring of 1919, the "Keeping Fit" campaign provides a unique insight into the federal government's attempt to medicalize and regulate American sexuality through the forum of public health. PMID- 12776437 TI - Prelude to the plague: public health and politics at America's Pacific gateway, 1899. AB - San Francisco played a crucial in the formulation of American immigration policy vis-a-vis Asia in the late nineteenth and early twentieth centuries. During this period, it was often difficult to differentiate political struggles over the exclusion of Asians from other conflicts. This article examines one such arena: an acrimonious, well-documented argument in 1899 between Federal and various State and local authorities over the arrival of a Japanese passenger liner that may--or may not--have been carrying bubonic plague. Six months later, the plague unquestionably arrived, resulting in the well-known San Francisco plague epidemic of 1900 in which more that 110 people died. Reviewing the 1899 prelude, the public attitudes of the various health authorities, and the way the press reported health issues, collectively give some sense of that historical space where the regulation of public health, politics, and the immigration industry intersected and were fiercely contested. PMID- 12776438 TI - Blood transfusion between the wars. AB - This article examines the introduction of blood transfusion into general practice from the end of the First World War to the Second World War. Developments during most of this period were not the result of new discoveries but rather the spread of ideas and the establishment of donor organizations to secure an adequate blood supply. The identification, testing, and organization of potential donors were done in a wide variety of settings that reflected differences in political and cultural experiences. At the end of the 1930s, with war approaching, the resolution of problems with storage of blood and the discovery of new techniques for separating and storing plasma dramatically changed transfusion practice. Thus, the innovations of the Second World War were very much based on the development of broad donor organizations plus the new technical discoveries that had occurred during the interwar period. PMID- 12776439 TI - Understanding asthma pathophysiology. AB - Asthma is best described as a chronic disease that involves inflammation of the pulmonary airways and bronchial hyperresponsiveness that results in the clinical expression of a lower airway obstruction that usually is reversible. Physiologically, bronchial hyperresponsiveness is documented by decreased bronchial airflow after bronchoprovocation with methacholine or histamine. Other triggers that provoke airway obstruction include cold air, exercise, viral upper respiratory infection, cigarette smoke, and respiratory allergens. Bronchial provocation with allergen induces a prompt early phase immunoglobulin E (IgE) mediated decrease in bronchial airflow (forced expiratory volume in 1 second) followed in many patients by a late-phase IgE-mediated reaction with a decrease in bronchial airflow for 4-8 hours. The gross pathology of asthmatic airways displays lung hyperinflation, smooth muscle hypertrophy, lamina reticularis thickening, mucosal edema, epithelial cell sloughing, cilia cell disruption, and mucus gland hypersecretion. Microscopically, asthma is characterized by the presence of increased numbers of eosinophils, neutrophils, lymphocytes, and plasma cells in the bronchial tissues, bronchial secretions, and mucus. Initially, there is recruitment of leukocytes from the bloodstream to the airway by activated CD4 T-lymphocytes. The activated T-lymphocytes also direct the release of inflammatory mediators from eosinophils, mast cells, and lymphocytes. In addition, the subclass 2 helper T-lymphocytes subset of activated T lymphocytes produces interleukin (IL)-4, IL-5, and IL-13. IL-4 in conjunction with IL-13 signals the switch from IgM to IgE antibodies. The cross-linkage of two IgE molecules by allergen causes mast cells to degranulate, releasing histamine, leukotrienes, and other mediators that perpetuate the airway inflammation. IL-5 activates the recruitment and activation of eosinophils. The activated mast cells and eosinophils also generate their cytokines that help to perpetuate the inflammation. Regardless of the triggers of asthma, the repeated cycles of inflammation in the lungs with injury to the pulmonary tissues followed by repair may produce long-term structural changes ("remodeling") of the airways. This review will discuss in greater detail the relationships of inflammation and airway hyperresponsiveness to the pathophysiology of asthma. PMID- 12776440 TI - Defining the effects of an inhaled corticosteroid and long-acting beta-agonist on therapeutic targets. AB - The effects of inhaled corticosteroids (ICSs) and long-acting beta 2-agonists (LABAs) on therapeutic targets have significant clinical relevance regarding optimal management of asthma. Asthma pathophysiology involves two main components: smooth muscle dysfunction and airway inflammation. LABAs and ICSs provide complementary modes of action in that these agents modulate smooth muscle dysfunction/bronchoconstriction and airway inflammation, respectively. Despite the documented benefits of ICSs, they remain underutilized because of a variety of physician- and patient-associated reasons including safety concerns. Underlying these concerns are published reports that suggest systemic effects of high doses of ICSs: skin bruising, reduction of bone mineral density, cataracts, glaucoma, and impaired short-term growth in children. Simple strategies to reduce the potential adverse effects of inhaled steroids include using the lowest effective maintenance dose and optimizing steroid-sparing strategies, specifically combination therapy with a LABA, leukotriene modifier, or theophylline. LABA therapy, when added to ICS therapy, provides clinically significant steroid-sparing effects while at the same time reducing the rate at which asthma exacerbations occur. Available clinical evidence suggests that the combination of ICS plus LABA is the best available option for the management of moderate persistent asthma. Consequently, this combination is the preferred choice for treating moderate persistent asthma based on current National Asthma Education and Prevention Program guidelines for the diagnosis and treatment of asthma. PMID- 12776441 TI - Clinical implications of combination therapy on the future of asthma management. AB - Current guidelines for the treatment of moderate persistent asthma list the combination of long-acting beta-agonists and inhaled corticosteroids (ICSs) as the treatment of choice. This decision is based on their efficacy compared with other dual controller combinations such as increased doses of ICSs or adding leukotriene modifiers, especially when the forced expiratory volume in 1 second value is used as the primary comparator. The main purpose of this study is to examine this form of dual controller therapy in terms of anti-inflammatory effects. One of the concerns with the combination of long-acting beta-agonists and ICSs is what happens to patients on this therapy during an asthma exacerbation. The existing data indicate that when combination therapy was compared with high-dose ICSs alone, it did not alter the ability to detect deteriorating asthma, it led to faster recovery after an exacerbation, it did not disguise deteriorating asthma, and it did not increase/mask airway inflammation. This study also examines how to approach patients placed on this dual controller therapy after control of their asthma is attained. Suggestions for the transition from initial therapy to maintenance therapy that may involve single controller medications are made. Innovative methods of handling asthma exacerbations with dual controller therapy also are made. PMID- 12776442 TI - Azelastine nasal spray: a review of pharmacology and clinical efficacy in allergic and nonallergic rhinitis. AB - Azelastine hydrochloride is a pharmacologically distinct H1-receptor antagonist with a broad spectrum of antiallergic and anti-inflammatory activity. Azelastine has established antiallergic and anti-inflammatory effects that are unrelated to H1-receptor antagonism, including inhibitory effects on the synthesis of leukotrienes, kinins, and cytokines; the generation of superoxide free radicals; and the expression of the intercellular adhesion molecule 1. Azelastine is available in the United States as a nasal spray formulation (Astelin) and is approved for treatment of seasonal allergic rhinitis and nonallergic vasomotor rhinitis. In U.S. clinical trials, azelastine nasal spray was effective in treating all of the symptoms of the allergic rhinitis symptom complex including ocular symptoms, and in double-blind clinical trials in nonallergic vasomotor rhinitis, azelastine nasal spray was effective in treating the total vasomotor rhinitis symptom complex including individual symptoms of nasal congestion and postnasal drip. This article reviews the pharmacologic profile and clinical efficacy and safety of azelastine nasal spray. PMID- 12776443 TI - Asthma or not? The value of flow volume loops in evaluating airflow obstruction. AB - Large airway obstruction can present with symptoms typically associated with asthma, including cough, dyspnea, or wheezing. Obstruction of the large airways should be considered in the differential diagnosis of patients with difficult to manage asthma, particularly those with known risk factors for tracheal disease, atypical clinical features, or lack of response to standard asthma therapy. Spirometry and flow volume loops are valuable diagnostic tools in this evaluation. This article discusses two cases in which large airway obstruction mimicked asthma and reviews the characteristic flow volume loop patterns associated with large airway lesions. PMID- 12776444 TI - Relationship between the tumor necrosis factor system and the serum interleukin 4, interleukin-5, interleukin-8, eosinophil cationic protein, and immunoglobulin E levels in the bronchial hyperreactivity of adults and their children. AB - The aim of the study was to investigate the activation of inflammatory mediators interleukin (IL)-4, IL-5, and IL-8; immunoglobulin E (IgE); and eosinophil cationic protein (ECP) and to evaluate the regulatory role of the tumor necrosis system (TNF) system in bronchial hyperreactivity. Adults who had suffered from bronchial asthma in childhood but who had been symptom free for at least 3 years were examined together with their children who did not have asthma. The serum concentrations of TNF-alpha, soluble TNF receptor 1 (sTNF-R1), TNF-R2, IL-4, IL 5, IL-8, ECP, and IgE were studied in symptom-free adults (n = 22) and their children (n = 22) with bronchial hyperreactivity. Nonhyperreactive individuals with a similar medical history (adults, n = 17; children, n = 20) served as controls. Significantly elevated serum TNF-alpha (X +/- SD: 5.13 +/- 1.37 pg/mL versus 3.91 +/- 0.61 pg/mL; p < 0.0001), sTNF-R1 (X +/- SD: 1.37 +/- 0.28 ng/mL versus 1.16 +/- 0.13 ng/mL; p = 0.0002), and sTNF-R2 (X +/- SD: 0.78 +/- 0.42 ng/mL versus 0.43 +/- 0.41 ng/mL; p = 0.0001); IL-4 (X +/- SD: 4.05 +/- 1.02 pg/mL versus 3.34 +/- 0.84 pg/mL; p = 0.0016); IgE (X +/- SD: 390.1 +/- 361.4 KU/L versus 130.2 +/- 166.1 KU/L; p = 0.0001); and ECP (X +/- SD: 17.57 +/- 11.03 micrograms/L versus 10.65 +/- 6.01 micrograms/L; p = 0.0016) concentrations were measured in the subjects with bronchial hyperreactivity as compared with the nonhyperreactive group. Significant positive linear correlations were observed for the bronchial hyperreactive group between the concentrations of TNF-alpha and ECP, TNF-alpha and sTNF-R1, TNF-alpha and IL-8, sTNF-R1 and ECP, sTNF-R1 and IL 8, and sTNF-R2 and IL-8. Moreover, the TNF-alpha and sTNF-R2 levels correlated with the airway reactivity in the hyperreactive group. We suggest that the elevated cytokine levels indicate activation of the immune system in individuals who were previously asthmatic, but recovered, and are now symptom free and in their children with nonasthmatic bronchial hyperreactivity. The TNF system may play a key role in the pathomechanism of bronchial hyperreactivity. PMID- 12776445 TI - The efficacy of nasal surgery among patients with and without allergies. AB - Surgical intervention has been used widely for the correction of nasal obstruction. However, there are only a small number of studies confirming its advantages. This study summarizes our experience regarding patients' satisfaction with the operation results. We also examined whether the presence of allergic rhinitis can be used as a reliable predictive criterion for the operation outcome in an adult population. Fifty-three patients with perennial nasal obstruction who had been referred for nasal surgery underwent allergic evaluation. Patients with a positive skin-prick test to a perennial allergen were included in the allergic group, whereas those with a positive skin test to a seasonal allergen only or with negative skin tests were included in the nonallergic group. Patients filled out a symptom score before the operation and at 6 and 12 months after the operation and also were asked to grade their overall satisfaction with the outcome of the operation. One year after surgery, only 20 patients (38%) were definitely satisfied with its results. Twenty-eight patients (53%) were willing to undergo the same operation again had they known of its outcome in advance. There was no difference between the allergic and nonallergic groups regarding patient satisfaction or willingness to undergo the same operation again. We conclude that the allergic status of the patient is not an effective predictive criterion for the outcome of nasal surgery in patients with perennial nasal obstruction. The efficacy of this procedure is rather low and better preoperative criteria are required to improve its efficacy. PMID- 12776446 TI - Fatal reactions to hymenoptera stings. AB - Previous studies measuring immunoglobulin E (IgE) antibodies in postmortem sera for determining the cause of fatal anaphylaxis have reported only single cases or small groups. Recently, more attention has been paid to reports of patients with venom allergies who are negative by venom skin testing, by in vitro testing or by both tests. The aim of this study was to determine the reliability of postmortem specific IgE antibody testing in venom anaphylactic death and the range of antibody levels found. Radioallergosorbent testing was performed on sera from three groups: 51 anaphylactic deaths from insect stings, 20 anaphylactic/anaphylactoid deaths from food and drugs, and 31 control subjects. Results were analyzed by descriptive statistics, chi-squared test, and receiver operating characteristic curve. The sera in the sting death group were significantly different from the other two groups, which were not statistically different from each other. The maximal sensitivity of the radioallergosorbent test was 90% at 0.35 ng/mL, and the optimal sensitivity was 73% with a specificity of 86% at 0.54 ng/mL. IgE antibody could not be detected in 10% of the sting death sera and levels from 0.35 to 0.65 ng/mL were found in 24%. The level of specific IgE antibodies against venoms is not predictive of the severity of anaphylactic reaction. PMID- 12776447 TI - Budesonide turbuhaler delivered once daily improves health-related quality of life in adult patients with non-steroid-dependent asthma. AB - Budesonide inhalation powder via the dry-powder multidose inhaler Turbuhaler (budesonide Turbuhaler) has been shown to improve lung function and symptoms in adults with asthma. In this double-blind, placebo-controlled, multicenter trial, we evaluated the effects of once-daily budesonide Turbuhaler on health-related quality of life (HRQL) in 177 adults (aged 18-70 years) with non-steroid dependent asthma. Patients were randomized to receive budesonide Turbuhaler (400 micrograms) once daily or placebo for 12 weeks. HRQL was assessed at baseline and weeks 4 and 12 using the Asthma Quality of Life Questionnaire (AQLQ). In addition to assessment based on the four AQLQ domains (activity limitations, asthma symptoms, emotional function, and response to exposure to environmental stimuli), AQLQ overall scores were analyzed for both treatment groups. Compared with placebo, patients using budesonide Turbuhaler once daily had statistically significant (p < 0.001) improvements from baseline to weeks 4 and 12 in AQLQ overall scores. Statistically significant improvements from baseline to weeks 4 and 12 in all four individual domains also were observed in the budesonide Turbuhaler group compared with placebo. Differences between the two treatment groups in mean changes from baseline in AQLQ overall, asthma symptoms, and emotional function reached the level required for patients to achieve a minimal important difference of change (> or = 0.5 U) at week 12. A retrospective analysis of the data showed that approximately 70% of patients treated with budesonide Turbuhaler experienced a minimal important difference of change in AQLQ overall scores. Overall, improvements in AQLQ correlated significantly (p < or = 0.04) with improvements in forced expiratory volume in 1 second, morning peak expiratory flow measurements, asthma symptoms, and breakthrough bronchodilator use at the study end. Thus, patients with corticosteroid-naive asthma can experience improved HRQL when using budesonide Turbuhaler. PMID- 12776449 TI - Beyond Botox. PMID- 12776448 TI - Pet allergy: how important for Turkey where there is a low pet ownership rate. AB - Exposure and sensitization to allergens derived from cats/dogs have been shown to represent an important risk factor for allergic respiratory diseases. So far, there has not been any study exploring cat/dog sensitization and related factors in our geographic location. The aim of this study was to determine the sensitization to cats/dogs in a group of patients with rhinitis and/or asthma and to evaluate the relationship between current and childhood exposure and sensitivity to pets. Three hundred twelve consecutive subjects with asthma and/or rhinitis were included in the study and were asked to reply a questionnaire concerning past and current pet ownership and presence of pet-related respiratory symptoms. After performing skin-prick tests, subjects were allocated into three groups: group 1 (n = 103), subjects with nonatopic asthma; group 2 (n = 54), allergic rhinitis and/or asthma patients with pet allergy; group 3 (n = 155), allergic rhinitis and/or asthma patients without pet allergy. Pet hypersensitivity was detected in 54 of 209 atopic subjects (25.8%). There was no difference in the rates of past pet ownership among subjects with (29.6%) and without (23.8%) pet allergy. However, the ratio of current pet ownership was higher in atopic patients with pet allergy (16.6%) than in nonatopic subjects (2.9%; p = 0.02). The prevalence of sensitization to pets in current owners (42.8%) was higher than prevalence of sensitization in patients who never had a pet (22.6%; p = 0.002; odds ratio, 2.67) and who owned a pet at childhood (28.2%; p = 0.038; odds ratio, 1.9). Thirteen subjects (13/54; 24%) described respiratory symptoms when exposed to cats and/or dogs. Rate of past pet ownership was similar in symptomatic and asymptomatic subjects with pet allergy (30.7% versus 29.2%; p > 0.05). Rate of current per ownership was higher in symptomatic subjects than in asymptomatic subjects with pet sensitivity (38.4% versus 9.5%; p < 0.0001). Our data indicate that pet allergens have the potential to become an important source of indoor allergens in our population. Our findings also suggest that current pet ownership--but not childhood pet keeping--seems to be a risk for the development of sensitization to pets. PMID- 12776451 TI - Isn't that special. Louisiana hospital could be the first--and, if successful, probably not the last--to finance a new facility by selling bonds to doctors. AB - Lafayette (La.) General Medical Center is trying to raise funds for a $75 million heart hospital by selling bonds to doctors. If the sale is successful, a flood of similar deals could follow. The project's chief architect, Robert Rosenfield, left, says his firm already is working on 25 to 30 additional deals. What effect will this type of financial arrangement have on the healthcare industry? PMID- 12776452 TI - Battle on the bayou. La. among states battling over doc-owned facilities. PMID- 12776453 TI - Veteran inspectors eyed to succeed Rehnquist. PMID- 12776454 TI - Shedding new light. IRS proposes rules to clarify HMO exemption. PMID- 12776455 TI - Big brother wants nurses. Possibility of federal mandates worries hospitals. PMID- 12776456 TI - Teetering on the brink. Greater Southeast seeks help from D.C. mayor. PMID- 12776457 TI - An electronic emergency. We need to push the industry toward standardized digital medical records. PMID- 12776458 TI - The best route. Clinical pathways have become a habit at Children's Hospital in San Diego and the effort is paying off in quantifying quality and cutting costs. PMID- 12776459 TI - Still hard to beat. Doctors don't have the heart--or a clinical reason--to part with their old-fashioned stethoscopes. PMID- 12776460 TI - The role and importance of definitions and standards in healing research. PMID- 12776461 TI - Samueli Conference on Definitions and Standards in Healing Research: working definitions and terms. PMID- 12776462 TI - Bioenergy definitions and research guidelines. AB - A model for the functional and observable interrelation among the various components in a physical bioenergy system is presented. The analogy is made between electric circuits and electromagnetic interactions, and contact and noncontact bioenergy transfer. It is postulated that there exists some form of bioenergy that has the capacity to do work and that this energy behaves in a manner similar to electricity in that the physical concepts of electromotive force, current, and impedance have their equivalents in bioenergy. It is further postulated that these analogous components are related by an equivalent to Ohm's and other physical laws of electricity. This is extended to a conjecture that bioenergy healing is the transfer of information from a practitioner to a healee. Research guidelines for bioenergy measurements are presented, including basic measurement practices for electrical and electromagnetic systems through direct measurements and the use of indirect measurement experiments for detecting these or other forms of bioenergy transfer. The research guidelines are divided into 2 sections: those involving direct measurement of the physical electrical properties of a practitioner, in particular the difficulties associated with electrical measurements of extremely low-level signals outside of a Faraday shield or electromagnetic measurements outside of a radio frequency anechoic chamber; and those for conducting experiments in which the effects of bioenergy are being investigated on the healee or other target system without direct measurements of the means for bioenergy transfer. PMID- 12776463 TI - Distant healing intention: definitions and evolving guidelines for laboratory studies. AB - This paper provides definitions and a discussion of evolving guidelines for conducting research on the effects of distant healing intention (DHI) on living systems in the laboratory. We consider the relevance of DHI laboratory research to applied healing, special theoretical challenges, and other considerations that distinguish DHI research from other domains of laboratory science. Two sample protocols for investigating DHI are provided, one involving the human autonomic nervous system as the "target" of distant intention, and the other involving cell cultures. In essence, DHI differs from other alternative healing modalities because it postulates that mental intention alone can affect living systems at a distance, unbounded by the usual constraints of both space and time. This postulate challenges scientific assumptions that often go unexamined, including the nature of causality, the distinction between subjective and objective states, and the efficacy of double-blind protocols in controlling for experimenters' intentions. Previous laboratory research in this domain suggests that DHI effects warrant serious study, but most scientists and funding agencies are unaware of the evidence or the relevant literature. By following these evolving guidelines, researchers' designs and their ultimate publications will conform more closely to the quality standards expected by scientific journals, and such publications will in turn attract the attention of a broader range of scientists. This seems especially important for alternative healing research in general and for distant healing in particular; both realms enjoy broad public support but have largely eluded serious attention by mainstream science. PMID- 12776464 TI - Healing prayer outcomes studies: consensus recommendations. PMID- 12776465 TI - Standards for conducting clinical biofield energy healing research. PMID- 12776466 TI - Research guidelines for assessing the impact of the healing relationship in clinical nursing. PMID- 12776467 TI - Research guidelines for assessing the impact of healing relationships in clinical medicine. AB - This paper charts a course for assessing the impact of healing relationships in clinical medicine. The system of healing relationships is multidimensional, longitudinal, contextual, and emergent. In a new conceptual model, healing relationships are identified in terms of the conditions of healing intention, motivation, and information transfer, and in terms of the attributes of emotional engagement, mindfulness, and trust. Five components of quality in healing relationships--adaptability, cohesion, growth, caring-in-relation, and commitment -are noted, and the importance of timing, attunement, and cultural meaning systems are described. Communication, clinical method, caring, competence, and treatment characteristics are differentiated as mediating processes; expectancy and conditioning are positioned as antecedents of healing relationships. Multiple personal and contextual outcomes are addressed with a recommendation for assessing a minimal set of each, including symptom resolution, health status, sense of coherence, patient enablement, cost effectiveness, quality of care, efficiency, access, and healer satisfaction. A wheel of knowledge connects 3 ways of knowing--personal, connected, and objective--with appropriate methodology and research designs. Applying this wheel to the issue of assessing impact in healing relationships reveals the need for multiple methods, perspectives, and triangulations. A critical multiplist strategy is one means for advancing this area of research. A double-helix trial design is introduced, in which one strand consists of a standard quantitative approach and the other consists of qualitative methods. The 2 strands are bonded by the questions addressed and by the participants in the study. PMID- 12776468 TI - A systematic review of the quality of research on hands-on and distance healing: clinical and laboratory studies. AB - PURPOSE: To systematically review the quality of published experimental clinical and laboratory research involving hands-on healing and distance healing between 1955 and 2001. DATA SOURCES: Studies were identified through comprehensive literature searches on spiritual healing in MEDLINE, PSYCH LIT, EMBASE, CISCOM, and the Cochrane Library from their inceptions to December 2001. STUDY SELECTION: We selected published randomized, controlled trials of spiritual healing (hands on healing and distance healing) done in clinical and laboratory settings, all of which had been peer reviewed. DATA EXTRACTION: Independent quality assessment of internal validity was conducted on all identified studies using the comprehensive Likelihood of Validity Evaluation scale. Clinical and laboratory studies were analyzed separately and then subdivided into hands-on healing or distance healing interventions. RESULTS: A total of 45 laboratory and 45 clinical studies published between 1956 and 2001 met the inclusion criteria. Of the clinical studies, 31 (70.5%) reported positive outcomes as did 28 (62%) of the laboratory studies; 4 (9%) of the clinical studies reported negative outcomes as did 15 (33%) of the laboratory studies. The mean percent overall internal validity for clinical studies was 69% (65% for hands-on healing and 75% for distance healing) and for laboratory studies 82% (82% for hands-on healing and 81% for distance healing). Major methodological problems of these studies included adequacy of blinding, dropped data in laboratory studies, reliability of outcome measures, rare use of power estimations and confidence intervals, and lack of independent replication. CONCLUSIONS: When laboratory studies were compared to clinical studies in the areas of hands-on healing and distance healing across the quality criteria for internal validity, distance healing studies scored better than hands on healing studies, and laboratory studies fared better than clinical studies. Many studies of healing contained major problems that must be addressed in any future research. PMID- 12776470 TI - 7th annual alternative therapies symposium and exhibition medicine and the planet: the coming age of ecological medicine. March 26-29, 2003 Seattle, Washington. PMID- 12776469 TI - Privacy. PMID- 12776471 TI - Labeling and manufacturing standards proposed for all dietary supplements. PMID- 12776472 TI - Planning grant awarded to study CAM in treatment of migraines and lower back pain. PMID- 12776473 TI - Management of hyperlipidemia. PMID- 12776474 TI - Herbs and other dietary supplements: healthcare professionals' knowledge, attitudes, and practices. AB - CONTEXT: Herbs and other dietary supplements (H/DS) are frequently used by the public. They have significant health implications, yet little is known about health professionals' knowledge, attitudes, or clinical practices related to H/DS. DESIGN: Cross-sectional survey of clinicians prior to participation in an Internet-based educational program on herbs and dietary supplements. PARTICIPANTS: The 537 participants included 111 physicians (MD), 30 advanced practice nurses (RN), 46 pharmacists (PharmD), and 350 dietitians (RD). In addition to demographic information, participants were asked about their knowledge, attitudes, and practices related to H/DS. RESULTS: Most participants were involved in direct patient care (85%), in practice or on faculty (84%), and from outside our local institutions (76%); 66% reported receiving professional education about H/DS in the past year. There were statistically significant differences between professional groups, with RDs scoring better than others, but even their average scores were less than 60% of possible. The average score on knowledge questions was 10/20; the average confidence score was 4 out of 10 possible, and the average communication score was 1.4 out of 4 possible. Most respondents knew the most common clinical uses of echinacea and St. John's wort, and felt confident that they knew more than their colleagues about H/DS. Key deficits were in knowledge about adverse effects, confidence in reporting side effects, routinely communicating with patients about H/DS, and recording H/DS information in the medical record. CONCLUSIONS: Despite significant interest and previous training in H/DS, these clinicians had substantial room for improvement in knowledge, attitudes, and clinical practices about H/DS. Educational interventions and institutional policies are needed to improve the quality of patient care regarding H/DS, and such interventions should be rigorously evaluated to ensure that continuous improvements occur. PMID- 12776475 TI - Use of complementary and alternative medicine among the ethnic elderly. AB - OBJECTIVE: We sought to explore whether the elderly are high users of complementary and alternative medicine (CAM), and to determine which modalities they use. We also sought to describe patterns and positive predictors of CAM use among 3 ethnically diverse groups of community-residing elderly. DESIGN: A 7-page questionnaire was developed and translated into Spanish and Vietnamese. PARTICIPANTS: A population of 525 community-residing elderly completed personal interviews. RESULTS: Two hundred and fifty-one respondents (47.8%) reported using CAM over the past year. Dietary supplements (47.4%), chiropractic (16.3%), home remedies (15.9%), acupuncture (15.1%), and Oriental medicine (12.8%), were the most frequently cited therapies. The majority of CAM users (62.4%) did not inform their physicians that they were using it, but 58% consulted their physician for the same problem for which they used CAM. Family and friends were most relied upon for making the choice of therapy. Among the 3 ethnic groups studied, Asians were higher users of acupuncture (28%) and Oriental medicine (31%), Hispanics were higher users of dietary supplements (56%), home remedies (25%), and curanderos (8%), while white non-Hispanics were higher users of chiropractic (42%), massage (20%), vitamins (20%), diet (17%), and psychospiritual (15%) modalities. Pain was a higher indicator of CAM use among Asians, gastrointestinal problems and diabetes among Hispanics, and stress/fatigue and cardiovascular problems among white non-Hispanics. CONCLUSION: Findings indicated a high use of CAM among the elderly and emphasize the likelihood that elderly immigrants use those therapies with which they are familiar. Modalities and conditions varied with the ethnicity of respondents. PMID- 12776476 TI - Long-term effects of dextrose prolotherapy for anterior cruciate ligament laxity. AB - CONTEXT: Use of dextrose prolotherapy. Prolotherapy is defined as injection that causes growth of normal cells or tissue. OBJECTIVE: Determine the 1 and 3 year efficacy of dextrose injection prolotherapy on anterior cruciate ligament (ACL) laxity. After year 1, determine patient tolerance of a stronger dextrose concentration (25% versus 10%). DESIGN: Prospective consecutive patient trial. SETTING: Outpatient physical medicine clinic. PATIENTS OR OTHER PARTICIPANTS: Eighteen patients with 6 months or more of knee pain plus ACL knee laxity. This laxity was defined by a KT1000 anterior displacement difference (ADD) of 2 mm or more. INTERVENTION: Intraarticular injection of 6-9 cc of 10% dextrose at months 0, 2, 4, 6, and 10. Injection with 6 cc of 25% dextrose at 12 months. Then, depending on patient preference, injection of either 10% or 25% dextrose every 2 4 months (based on patient preference) through 36 months. MAIN OUTCOME MEASURES: Visual analogue scale (VAS) for pain at rest, pain on level surfaces, pain on stairs, and swelling. Goniometric flexion range of motion, and KT1000-measured ADD were also measured. All measurements were obtained at 0, 6, 12 and 36 months. RESULTS: Two patients did not reach 6 month data collection, 1 of whom was diagnosed with disseminated cancer. The second was wheelchair-bound and found long-distance travel to the clinic problematic. Sixteen subjects were available for data analysis. KT1000 ADD, measurement indicated that 6 knees measured as normal (not loose) after 6 months, 9 measured as normal after 1 year (6 injections), and 10 measured as normal at 3 years. At the 3 year follow-up, pain at rest, pain with walking, and pain with stair use had improved by 45%, 43%, and 35% respectively. Individual paired t tests indicated subjective swelling improved 63% (P = .017), flexion range of motion improved by 10.5 degrees (P = .002), and KT1000 ADD improved by 71% (P = .002). Eleven out of 16 patients preferred 10% dextrose injection. CONCLUSION: In patients with symptomatic anterior cruciate ligament laxity, intermittent dextrose injection resulted in clinically and statistically significant improvement in ACL laxity, pain, swelling, and knee range of motion. PMID- 12776478 TI - The effectiveness of Commiphora mukul for osteoarthritis of the knee: an outcomes study. AB - CONTEXT: Ayurveda, the traditional system of healthcare in India, has many remedies for Osteoarthritis (OA). One of the ingredients most commonly found in Ayurvedic arthritis formulas is guggul, an oleoresin of the herb Commiphora mukul (CM). The authors have conducted both preclinical and clinical investigations of guggul for reduction of pain, stiffness, and improved function, and to determine tolerability in older patients with a diagnosis of OA of the knee. METHODS: The study was conducted using an outcome, quasi-experimental, model. Thirty male and female participants meeting the inclusion/exclusion criteria, with a score of 2 or more on the Kellegran-Lawrence scale for at least 1 knee, were admitted in the study. CM was administered in capsule form (500 mg concentrated exact delivered TID) along with food. The WOMAC Total Score was used as a primary outcome measure. VAS scales, 6-minute walk-test, and WOMAC subscales were used as outcome measures. RESULTS: At the end of treatment, there was a significant difference in the scores of the primary and secondary outcome measures. On the primary measure, WOMAC total score, participants were significantly improved (P < 0.0001) after taking the supplement for 1 month and continued to improve at the 2-month marker and follow-up. Secondary measures of pain in the VAS format demonstrated participant improvement; however, mood state, and current pain were not significantly different (P < 0.05) than baseline until the 2 month assessment (P < 0.001). CONCLUSIONS: Overall data indicate significant improvement for participants during the trial in both scales and objective measures used for assessment purposes. There were no side effects reported during the trial. CM appears to be a relatively safe and effective supplement to reduce symptoms of OA. PMID- 12776477 TI - Religious and spiritual resources, CAM, and conventional treatment in the lives of cancer patients. AB - CONTEXT: In addition to seeking conventional treatment from physicians, cancer patients will often use religious and spiritual resources (RSR) and complementary and alternative medicine (CAM). Patients' beliefs about the relationships among RSR, CAM, and conventional treatments may reflect belief systems not readily apparent to physicians. OBJECTIVE: 1) Identify the RSR used and explore themes in beliefs regarding RSR, CAM, and conventional treatment. 2) Investigate the nature of the relationships among RSR, CAM, and conventional treatment in the lives of cancer patients. DESIGN: Cancer patients in Hawaii were recruited from a group who had previously completed a questionnaire on CAM use. In-depth interviews were conducted with a selected subset of survey participants. PARTICIPANTS: 143 cancer patients were interviewed 2 to 3 years following diagnosis. RESULTS: Participants reported using a variety of RSR, including personal faith, individual (self) prayer, relationship/dialog with God, prayers from fellow church members and others, counseling from pastor/priest or leader of faith, reading the bible, attending religious services, meditation, finding and spending time at locations of spiritual energy (i.e., churches, specific geographical locations, or certain natural settings), and help or counseling by ancestor(s). They also expressed beliefs that RSR, CAM, and conventional treatment have both shared and distinct purposes. RSR themes included providing treatment or cure, supporting treatment or cure, and being part of life. CAM themes included providing treatment or cure, supporting treatment or cure, preventing cancer and recurrence, substitution for conventional treatment, and as a last resort. Participants believed the purposes of conventional treatment were to fight/cure cancer and to improve quality of life. CONCLUSIONS: RSR, CAM, and conventional treatment served both spiritual and physical purposes. When participants perceived a relationship between RSR, CAM, and conventional treatment, it reflected shared spiritual and physical purposes. Such associations were especially strong when RSR, CAM, and conventional treatment shared the common goals of providing treatment or cure and/or supporting treatment or cure. Perceptions of shared purpose were individual to each participant, with religious congruence and life history playing a role in beliefs about the relationship between RSR and CAM. Such beliefs can affect issues of control, spiritual well-being, coping, depression, decision-making, and possibly health outcomes in cancer patients. Therefore, they should be addressed by clinicians in discussions with their patients. PMID- 12776479 TI - Ken Cohen, MA, MSTh. Healing through ancient traditions: Qigong and Native American medicine. Interview by Bonnie Horrigan. PMID- 12776480 TI - Vitamin C as a cancer treatment: state of the science and recommendations for research. PMID- 12776481 TI - Progress notes: University of California, San Francisco. PMID- 12776482 TI - Stealing the soul, soumwahu en naniak, and susto: understanding culturally specific illnesses, their origins and treatment. PMID- 12776483 TI - The Healing Arts Center at St. John: connecting CAM into the traditional hospital setting. PMID- 12776484 TI - Allergens, dysbiosis and immune dysregulation: case studies on inflammatory bowel disease. PMID- 12776485 TI - The importance of small research projects: the impact of full text online journals on journal use surveys. PMID- 12776486 TI - The College of Physicians of Philadelphia's Regional Community Health Information System Project. PMID- 12776487 TI - JCAHO's new initiative. PMID- 12776488 TI - Survival of the fittest. PMID- 12776489 TI - Why men die young. There's a reason women are healthier, and it's not entirely in their genes. PMID- 12776490 TI - A working Mother's Day, from A to Z. PMID- 12776491 TI - [Conservation of endangered species resources of Tibetan medicine in China]. AB - OBJECTIVE: To further investigate and discuss the cause of species endangerment, the status and present problem of conservation of traditional Tibetan medicine in China. METHOD: Previous relevant investigations and literatures were summed up in the field. The present situation of conservation of traditional Tibetan medicine was analyzed. RESULT: The status of endangered resources, cause of species endangerment, the conserving status and conserving measures etc were elaborated. The classification was made and suggestion of species conservation of traditional Tibetan medicine were put forward. CONCLUSION: The endangered species conservation of traditional Tibetan medicine was carried out by building protective area of endangered species resources and plant garden, setting up germplasm bank, developing the domestication and cultivation of Tibetan medicinal herbs most in use, strengthening the investigation and study of endangered species, launching exchange and cooperation of conservation techniques on endangered species, enhancing the protective awareness of endangered species traditional Tibetan medicine etc. By so doing we can facilitate the sustainable development of traditional Tibetan medicine in China. PMID- 12776492 TI - [Studies on the pharmacological effects of saffron(Crocus sativus L.)--a review]. AB - OBJECTIVE: Reviewing the studies on the chemical components and medicinal value. METHOD: Philological method. RESULT: Saffron is a conventional effective medicine in improving blood circulation and curing the bruise. The late of evidesnces indicate that saffron possesses anticancer activity against a wide spectrum of tumors, such as leukemia, ovarian carcinoma, colon adenocarcinoma, rhabdomyosarcoma, papilloma, squamous cell carcinoma, and soft tissue sarcoma. It has low biochemical toxic effects on animals. In addition, saffron can be used to cure coronary heart disease and hepatitis, and to promote immunity. CONCLUSION: Saffron is a highly valuable medicine. And producing it in a large quantity has a wide application prosperity. PMID- 12776493 TI - [Research on the identification of penis et testis cervi with molecular taxonomy]. AB - OBJECTIVE: To make the kit with witch to identify Penis et Testis Cervi with molecular taxonomy. METHOD: The mtDNA of sika and red deer from different areas was amplified by PCR and sequenced. Compared with the mtDNA of bovine and horse from witch the false medicines were made, characteristic segments of deer were found. We selected one as the species distinctive PCR primer of deer. RESULT: The kit made up with this primer and related reagents could be used to discern Penis et Testis Cervi from the false medicine. CONCLUSION: It is a scientific, steady, accurate and convenient way to identify Penis et Testis Cervi with molecular taxonomy. PMID- 12776494 TI - [A novel approach to quality evaluation of root of Scutellaria baicalensis by DPPH free radical scavenging]. AB - OBJECTIVE: To develop a simple, reliable approach for evaluating the quality of Huangqin (Scutellaria baicalensis). METHOD: To determine the DPPH free radical scavenging activity and assay of four bioactive components: baicalin, baicalein, wogonin and wogonin-7-O-glucuronide by HPLC. RESULT: The correlative relationship between DPPH free radical scavenging activity and baicalin content was obtained. CONCLUSION: Bioassay of DPPH free radical scavenging activity could be used as one of the methods for quality evaluation of Chinese drug Huangqin. PMID- 12776495 TI - [Studies on the chemical constituents from the roots and rhizomes of Gerbera piloselloides]. AB - OBJECTIVE: To study the chemical constituents from the roots and rhizomes of Gerbera piloselloides. METHOD: The compounds were isolated by silica gel column chromatography, and their structures were elucidated by means of spectral analysis. RESULT: Seven compounds were identified as bothrioclinin (I), hydroquinone (II), marmesin (III), succinic acid (IV), umbelliferone (V), isoarborinol (VI) and beta-sitosterol (VII). CONCLUSION: Compound I, V and VI were isolated from Gerbera genus for the first time. PMID- 12776496 TI - [Studies on chemical constituents in fruit of Eucalyptus globulus]. AB - OBJECTIVE: AIM To study the chemical constituents of the fruit of Eucalyptus globulus. METHOD: Various column chromatographies with silica gel were employed for the isolation and purification. The structures of the compounds were elucidated with of spectral analyses and chemical methods. RESULT: Five compounds were isolated and elucidated as follows: betulonic acid (I), betulinic acid (II), ursolic acid (III), corosolic acid (IV), daucosterol (V). CONCLUSION: Compound I, II, III and IV were isolated from this plant for the first time. PMID- 12776497 TI - [Chemical constituents in volatile oil from the flos of Dendranthema indicum var. aromaticum var. nov]. AB - OBJECTIVE: To analyse chemical constituents of volatile oil from Dendranthema indicum var. aromaticum var. nov. by GC-MS. METHOD: The volatile oil was extracted from Dendranthema indicum var. aromaticum var. nov. through steam distillation and was analysed with different kinds of capillary columns to find out the optimal conditions. The content of compositions of volatile oil was determined with normalization method, and the constituents were identified by GC MS. RESULT: 44 Components were separated and identified, which accounted for over 43% of total volatile oil. CONCLUSION: The main constituents in the essential oils from Dendranthema indicum var. aromaticum var. nov. are Verberol, ( )zingiberene, beta-sesquiphellandrene, farnesene, transchrysanthenyl acetate, and caryophyllene. PMID- 12776498 TI - [Investigation on NB4 cell responses to realgar by cDNA microarray]. AB - OBJECTIVE: To elucidate the molecular mechanism of realgar-induced apoptosis and differentiation of acute promyelocytic leukemia(APL) cell line NB4. METHOD: The response of NB4 cells to realgar was explored with a cDNA microarray representing 1003 different human genes. RESULT: The analysis of gene expression profiles indicated that 9 genes were up-regulated and 37 genes were down-regulated. Among the 9 up-regulated genes, 2 genes were involved in proteasome degradation pathway. CONCLUSION: PSMC2, PSMD1 and ITGB1 genes may play a role in the apoptosis and differentiation of NB4 cells. PMID- 12776500 TI - Biotech's billion dollar breakthrough. PMID- 12776499 TI - [Effect of dan-shen, san-qi of different proportion on platelet aggregation and adhesion in normal rabbits]. AB - OBJECTIVE: To observe the effect of the crude extracts of Dan-shen root and San qi of different proportion on platelet aggregation and adhesion in normal rabbits. METHOD: With rabbits, ig. (4d, exsanguinated via carotid artery, percentage of platelet aggregation and adhesion was measured. RESULT AND CONCLUSION: Aspirin (4.4 mg/Kg) could markedly inhibit platelet aggregation and adhesion in normal rabbit. The proportions of Dan-shen root/San-qi (10:0, 10:1, 10:3, 10:6, 1:10) could markedly inhibit platelet aggregation, among which 10:3 was the best. San-qi alone had little effect on aggregation. The proportions of Dan-shen root/San-qi (10:3, 10:6, 0:10) could markedly inhibit platelet adhesion, among which 0:10 was the best, and the proportions(10:0, 10:1, 1:10) had little effect. PMID- 12776501 TI - [Cancer of the upper stomach: current problems of its diagnosis]. AB - Data on 1248 cases of stomaches radically operated on for cancer at the surgical departments of the Moscow Regional Research Clinical Institute in 1971 to 2000 were used to study a number of problems associated with cancer of the upper stomach, by regarding the present-day role of radiation diagnosis as of paramount importance. The following radiation diagnostic techniques were assessed. Among them there were traditional X-ray studies (in each case), the new radiation diagnostic techniques: ultrasound study and computed tomography (CT) (750 studies), magnetic resonance imaging (MRI) (120 studies). The potentialities of radiation diagnosis and endoscopy performed in all cases are compared. A specially developed procedure for exploring endophytic forms of gastric cancer was employed in the morphological studies of the material. The incidence of cancer of the upper stomach and its association with the esophagus are presented; some aspects of the morphogenesis of cancer of this site are discussed. The relationship of its clinical symptoms and its radiation image has been studied. In the authors' opinion, the past two decades' rise in the incidence of proximal gastric cancer is one of the main problems in diagnosing gastric cancer that continues holding its stand in the general structure of cancer morbidity. The tendency for diffuse and mixed forms to increase in the morphogenesis of gastric cancer provides evidence that radiation techniques should be actively used in its diagnosis. It is necessary to apply classical double-contrasting X-ray study and endoscopy on equal grounds as basic diagnostic methods at early stages. Only their concord use may change the poor situation associated with the diagnosis of cancer at this site. CT and MRI as additional techniques may substantially provide more required diagnostic information. This is first and foremost associated with difficulties in gastric endoscopic study when esophageal cancer is apparently proved. PMID- 12776502 TI - [X-ray study of patients at the stage of preparation for prosthesis by using dental maxillary implants]. AB - The paper deals with the possibilities of using the data of X-ray studies- computed tomography and orthopantomography--in planning the surgical stage of implantation for preprosthetic correction of the upper denture. Examination of 23 patients has yielded the data showing what estimation of the size of an image obtained by the "Denta Scan" programme provides. PMID- 12776503 TI - [Use of current radiation techniques in the diagnosis and treatment of juvenile angiofibromas of the nasopharynx]. AB - In 1992-2001 seventeen patients with juvenile nasopharyngeal angiofibromas (JNAs) were treated at Krasnoyarsk Territorial Clinical Hospital. The patients underwent a comprehensive diagnosis involving computed tomography (CT), magnetic resonance imaging (MRI), and angiography, followed by tumor vascular embolization. Later on JNAs were surgically removed in 14 patients, a course of radiation therapy was performed. In suspected JNA, the authors consider it necessary to concomitantly use CT, MRI, and angiography, which provides the most complete diagnostic picture of a pathological focus. X-ray endovascular occlusion considerably lowers blood loss, facilitates a complete angiofibroma removal, and affects the tolerability of radiation therapy in inoperable cases. PMID- 12776504 TI - [To the role of computed tomography in the algorithm of diagnosis of lung cancer]. AB - The paper summarizes the results of examination of 100 patients with malignant tumors of the lung. The results of X-ray study, linear tomography, and computed tomography (CT) used in the examination of patients with lung cancer were compared by using as an indicator the following parameters: to make diagnosis of lung cancer, to reveal mediastinal intergrowths, to detect enlarged lymph nodes, pulmonary metastases, and pleural lesions, to obtain information on tumor spread into the chest, to reveal growths into the vascular walls. To evaluate the efficiency of the techniques in solving the above diagnostic tasks, a statistical analysis including the estimation of their accuracy, specificity, and sensitivity was made. Recommendations for examination of patients with lung cancer were drawn up for therapeutic institutions of different types. PMID- 12776505 TI - [Results of coronary stenting in relation to the length of an implanted stent]. AB - The paper considers the impact of the length of an implanted stent on the early and late outcomes of intracoronary stenting. The study included 177 patients on a retrospective basis. They all underwent intracoronary stenting of 215 stenoses. All the patients were divided into three groups in relation to the length of an implanted segment (under 20 mm, 20-30 mm, and under 30 mm). In 16 (9.04%) of the 177 patients, for complete stenosis correction, several, but not one, stents were inserted, i.e. multiple stenting of a coronary stenting was performed. The stents were implanted by the conventional procedure. There was no significant difference in clinical and angiographic success rates between the patients of all three groups. There were no complications, such as transmural myocardial infarction and deaths in any groups. Acute stent thrombosis was noted only in 1 (1.8%). There was no significant difference in the incidence of subacute stent thrombosis between Groups 2 and 3 patients. No case of subacute thrombosis of implanted stents was noted in Group 1 patients. Thus, acute and subacute stent thromboses were observed in 4 (2.3%) patients, they being all from Groups 2 and 3. There was no significant difference in the development of restenosis between Groups 1 and 2 and Groups 2 and 3. At the same time, restenosis significantly more frequently developed in patients from Group 3 than in those from Group 1. Thus, the findings indicate that the length of a stented segment is an important factor that influence both early and late outcomes of intracoronary stenting. PMID- 12776506 TI - [Potentialities of computed tomography in the diagnosis of hiatal hernia]. AB - An analysis of the results of 98 studies has demonstrated the potentialities of X ray computed tomography in the diagnosis of hiatal hernia (HH), provided the X ray computed tomography (XCT) semiotics of HH. It has found that XCT may directly visualize and objectively evaluate anatomic structures, such as diagraphmatic crus and esophageal foremen. It has emphasized that when XCT of abdominal and thoracic organs is performed, it is necessary to include the areas of diagragmatic curs and esophageal foramen into the list of anatomic structures binding for visual assessment and characterization, which in combination with other studies will assist in the early diagnosis of hiatal hernia and eventually expand the potentialities of XCT to a greater extent. PMID- 12776507 TI - [Arthrosonography in the diagnosis of rheumatoid arthritis of the knee joint]. AB - The paper presents the capacities of echography in the estimation of the activity of rheumatoid gonitis. A study of knee joints in 126 patients with rheumatoid arthritis has provided the basic ultrasound criteria that are characteristic for the first, second, and third degrees of its progression, which include the pattern of proliferative changes in the synovium, the qualitative and quantitative assessments of intraarticular exudate and its viscosity. The capacities of X-ray study and MRI in the detection of the progression of rheumatoid arthritis are discussed. An algorithm of knee joint radiation study is proposed for patients with rheumatoid arthritis. PMID- 12776508 TI - [X-ray densitometry of the axial skeleton and ultrasound densitometry of the calcaneus: potentialities and values of the techniques in clinical practice]. AB - Double-power X-ray absorptiometry of the axial skeleton and ultrasound densitometry of the calcaneus were used to examine 3 groups of patients: 58 with chronic renal failure (CRF), 73 with senile osteoporosis, and 78 with diabetes mellitus (DM). The examination has indicated that X-ray absorptiometry is the most informative method in diagnosing osteoporosis in patients with CRF and DM. Ultrasound densitometry is of informative value only in patients with senile osteoporosis and it may be used as a screening tool in this category of patients. PMID- 12776509 TI - [The setting-up of the Russian Association of Mammologists]. PMID- 12776510 TI - [Clinical use of the ultrasound contrast Levovist for hepatic graft visualization]. PMID- 12776511 TI - [Problems of and suggestions to the study of Chinese phytomedicine]. AB - OBJECTIVE: To provide reference material for the adaptation of the study on Chinese phytomedicine to the global research and development of phytomedicine. METHOD: Based on research experiences, some problems, such as resources, quality and effective components of Chinese phytomedicine, were discussed, and some suggestions were put forward. RESULT AND CONCLUSION: Facing the opportunity brought by the global research and development of phytomedicine, it is essential to solve several main problems for the global recognition of Chinese phytomedicine. PMID- 12776512 TI - [Review of germplasm resources studies on Salvia miltiorrhiza]. AB - Research development of germplasm resources of Salvia miltiorrhiza was reviewed in the aspects of species resource, geographic distribution and varieties, biological characters and culture varieties, and cell and tissue culture. It was suggested that more attention be paid to waters-soluble constituents when evaluating the quality of the medicine, and much more basic studies be done in the course of establishing standard plant base for high quality herb and in the aspect of new variety selection. PMID- 12776513 TI - [Field test and lab experiment on control efficacy of the pathogen of opium poppy mildew]. AB - OBJECTIVE: To screen effectual fungicides for field control because of the seriousness of opium poppy mildew and importance of chemical control on plant diseases. METHOD: Seven fungicides were screened in Lab experiment and field test during 1996-1997. RESULT AND CONCLUSION: All of them and their different dosages were effective to control conidia of Peronospora arborescens. Among them, 72.2% propamocarb of 1203 and 902.5 ppm were the most effective both in Lab experiment and field test with efficacy 79.91% and 79.33% respectively in field test, and the efficacy of other fungicides was over 50%. Seven fungicides tested can be used to control nonsystematic symptom of opium poppy mildew. PMID- 12776514 TI - [RAPD analysis on different varieties of Rehmannia glutinosa]. AB - OBJECTIVE: To provide molecular evidences for its breeding by studying the genetic relationship among varieties of Rehmannia glutinosa. METHOD: Nineteen varieties were detected by Randomly Amplified Polymorphic DNA(RAPD) markers. RESULT: The 20 selected primers produced 163 bands, among which 114(69.9%) were polymorphic. A DNA molecular dendrogram was established based on Hierarchical cluster analysis of 163 DNA bands amplified by 20 primers, which divided the 19 varieties into four groups: Group Beijing, Group 85-5, Group Guolimao and the other Group. CONCLUSION: 8 varieties of Group Beijing have a close genetic relationship, and so have varieties of Group 85-5, which provides information for Rehmannia glutinosa's breeding. PMID- 12776515 TI - [Study on improvement of dissolution rate of water-honeyed pills of six herbs with rehmunnia by technique of super fine crushing]. AB - OBJECTIVE: To evaluate the affection of crushing technology on quality. The dissolution of Pills of Six Herbs with Rehmunnia prepared by different crushing technology was determined by taking the dissolution of Paeonol as test marker. METHOD: The Pills was prepared with the fine powder which was crushed by normal crusher or super fine crusher. The rotatory-basket method was used, and the cumulative dissolution percentage was determined by UV. RESULT: Statistics indicated there was a significant difference in dissolution parameter (T50) between super fine crushing powder Pills and normal fine-crushing powder Pills (P < 0.01), and there was a difference in dissolution of different batches of Pills of Six Herbs with Rehmunnia prepared by the normal crush technique (P < 0.05). CONCLUSION: The determination of dissolution of Pills of Six Herbs with Rehmunnia is necessary. In order to improve the quality of drugs, we should adopt the technique of super fine crushing in the preparation procedure. PMID- 12776516 TI - [Studies on chemical constituents of the rhizomae of Ligusticum chuanxiong]. AB - OBJECTIVE: To study the chemical constituents of Ligusticum chuanxiong. METHOD: The constituents were separated and purified by column chromatography with silica gel and ODS, and identified by NMR, MS spectral data. RESULT: Eight compounds were isolated and identified as Ligustilide(I), 4,5-dihydro-3-butylphthalide(II), Z, Z'-6,6',7,3 alpha-diligustili-de(III), Z-6,8',7,3'-diligustilide(IV), Z'-3,8 dihydro-6,6'7,3' alpha-diligustilide(V), 4-hydroxy-3-butylphthalide(VI), ferulic acid(VII) and Xiongterpene(VIII). CONCLUSION: Compound VIII is a new compound. PMID- 12776517 TI - [Comparative study on anti-hypercholesterolemia activity of diosgenin and total saponin of Dioscorea panthaica]. AB - OBJECTIVE: To compare the anti-hypercholesterolemic and cholesterol absorption inhibitory activities between total saponin of Dioscorea panthaica (TSDP) and diosgenin (Dio). METHOD: TSDP and Dio were given ig or i.p. to mice or rats treated with cholesterol feed to evaluate their preventive and therapeutic effect on hypercholesterolemia. TSDP or Dio and cholesterol were mixed with pig bile to form the micelle, then the freeing cholesterol was detected to evaluate inhibitory effect of the both compounds on cholesterol absorption. RESULT: Dio (80 and 160 mg.kg-1) showed significantly therapeutic and preventive effect on hypercholesterolemia in mice, while TSDP showed a certain preventive activity only at a big dose (400 mg.kg-1). The intraperitoneal injection of Dio (20 and 40 mg.kg-1) to mice suffered from hypercholesterolemia was effective, but TSDP showed no effective. The serum total cholesterol level was decreased when rats were pre-treated with TSDP (200 and 400 mg.kg-1, ig) and Dio (200 and 100 mg.kg 1, ig). However, the hypercholesterolemia-preventing activity of Dio was stronger than that of TSDP. In addition, inhibitory effect of Dio on cholesterol micelle formation was still stronger than that of TSDP. CONCLUSION: The preventive and therapeutic activity of Dio against hypercholesterolemia indused by cholesterol in mice or rats is stronger than that of TSDP. The anti-hypercholesterolemia mechanism of Dio is probably related with its cholesterol absorption inhibitory activity. PMID- 12776518 TI - [Effect of zishuijianghuoyin on expression of lymphocyte AngII receptor (AT-1) mRNA of experimental hypertension rats]. AB - OBJECTIVE: To explore the immuno-neurologic regulation of hypertension and its inherent law as well as the mechanisms of curing and systemic regulating effect of ZiShuiJiangHuoYin(ZSJHY). METHOD: To detect expression level of AT-1mRNA in two-kidney-one-clamp renal hypertension rat lymphocyte cell by means of RT-PCR. RESULT: The level of AT-1 mRNA in lymphocyte was higher in group of 2K1C-RHR than that in normal group. ZSJHY (20 g/kg, 40 g/kg) had regulating effect on this. CONCLUSION: Depressing excessive expression of lymphocyte AT-1 mRNA may be one of the mechanisms where ZSJHY exert immunoregulation action. PMID- 12776519 TI - [Variation of endogeneous ET, CGRP and NO in myocardial ischemia in rats and the regulatory effect of xinshuping]. AB - OBJECTIVE: To observe the variation of ET and CGRP contents in ischemic heart, and NO level in serum of myocardial damaged rats, and their regulation when with the protection of Xinshuping, a traditional Chinese medicine compound. METHOD: The models of myocardial ischemia were prepared by subcutaneous injection of isoproterenol or by ligation of coronary artery. RESULT: ET content in myocardium was significantly increased (P < 0.01), and CGRP content as well as NO level in serum was not changed obviously in the model induced by isoproterenal. However, NO level in serum of rats treated with Xinshuping (ig bid x 2.5 d) was markedly raised (P < 0.01), neither ET not CGRP contents were affected by it. LDH and CK levels in serum of rats were evidently lowered by Xinshuping treatment. S-T segment's elevation of ECG was significantly inhibited and myocardial infarction size was reduced markedly by Xinshuping treatment in rats subjected to coronary artery ligature. CONCLUSION: ET, CGRP or NO is involved in myocardial infarction caused by isoproterenol. The ischemic damage or dysfunction in different models is obviously protected by Xinshuping. The promotion of NO release from vascular endothelium is probably related with this protective effect. PMID- 12776520 TI - [Mechanical research on effects of yishenqing on membranous nephropathy]. AB - OBJECTIVE: To investigate the therapeutic effect and mechanism of Yishenqing on rabbit membranous nephropathy. METHOD: The rabbit membranous nephropathy model was induced by cationic bovine serum albumin, and the immunity of mice together with urine volume and haemorheological property of rats were also estimated. RESULT: Yishenqing could significantly reduce the urinary protein content, preserve the renal function and pathologically restore the glomeruli. Moreover, its effects also include immunity enhancement, diuretic property and blood stasis amelioration. CONCLUSION: Yishenqing has good effects on the rabbit membranous nephropathy, which is probably due to the effects of diuretic property, immunity enhancement and blood stasis amelioration. PMID- 12776521 TI - [Studies on immunoregulating effect of monkshood root and peony root used singly and in combination]. AB - OBJECTIVE: To investigate the abirritation and antiinflammation effects of Monkshood Root and Peony Root used singly and in combination, and to find the enhanced effects of the two drugs used in combination; To observe the effect of Monkshood Root and Peony Root used singly and in combination by studing the immunoregulation function in experimental animals. METHOD: The response of delayed type hypersensitivity in mice, the phagocytosis of abdominal macrophages in mice, and the production of special antibodies in mice were observed. RESULT: The two drugs used in combination could increase phagocytosic function of mononuclear macrophagocyte in hypoimmuitic model mice, and inhibit the responses of delayed type hypersensitivity in the hyperimmunitic model mice and the nonimmunosuppressive treated mice, with nosignificant effect on the production of special antibodies in mice. CONCLUSION: In accordance with the mechanism of the disorder between the anti-inflammation effect and the induce-inflammation effect on arthritis in the immune system, these data show the bidirectional effect of the two drugs used in combination on the immune responses, which may be one of the main mechanisms of treating arthralgia due to wind-dampness. PMID- 12776522 TI - [Survey of pharmacology of oleanolic acid]. AB - OBJECTIVE: To summarize the pharmacological function of oleanolic acid. METHOD: Based on the documents and related achievements, the pharmacological function and its mechanism including its clinical use were summed up. RESULT AND CONCLUSION: Oleanolic Acid is a natural chemical which has wide-ranging function and abundant source. But we should place the emphasis on improving its biotic utilization and effect on clinical treatment. PMID- 12776523 TI - [Somatic hybridization between carrot and Bupleurum scorzonerifolium and esteras isoenzyme analysis]. AB - OBJECTIVE: To transfer the effective elements of Bupleurum scorzonerifolium into carrot, and provide theoretical data for the exploitation, improvement and selection of the germplasm of Chinese medicinal plants. METHOD: The protoplasta of Bupleurum scorzonerifolium irradiated by ultraviolet light (UV) at an intensity of 300 microW.(cm2)-1 for 0, 1, 2 min respectively were fused with those of carrot Fisch by PEG method. The regenerated clones, derived form a single fused cell, were examined for their hybrid nature by phenotype and Esterase isoenzyme analysis. RESULT: Nine clones were identified as the somatic hybrids between B. scorzonerifolium and carrot. CONCLUSION: This provides a firm foundation for the further analysis of the main active components saikosaponin of somatic hybrids and the screening out of high-medicine-content hybrid cell lines. PMID- 12776524 TI - [Determination of the content of saponin in Kochia scoparia fruits from different producing areas]. AB - OBJECTIVE: To compare the content of momordin Ic and total saponin in K. scoparia fruits from eleven producing areas. METHOD: HPLC-ELSD and colorimetric method were used to determine the content of momordin Ic and total saponin, respectively. RESULT: The content of momordin Ic in K. scoparia fruits was related to that of total saponin. In the eleven kinds of K. scoparia fruits tested, those produced in Bozhou, Baoding Anguo and Heilongjiang contained more saponins. CONCLUSION: The content of saponin in K. scoparia fruits from various areas is different, and attention must be paid to the effects of environment on the quality of herbs. PMID- 12776525 TI - [Study on the biology of adults parasite of Cordyceps sinensis, Hepialus biruensis]. AB - OBJECTIVE: To supply the basis of the breeding the host of parasite of Cordyceps sinensis for it's natural culturing, by studying the biology of the host of parasite of C. sinensis, Hepialus biruensis. METHOD: The host of parasite was collected in field, eclosion, mating and spawning in the different conditions were made, and it's biology and the regularity of the growth and development were observed. RESULT AND CONCLUSION: In the natural condition, H. biruensis has the rate of eclosion, 42.3%, and the sex ratio, 1:0.86, and it's lifespan is 2-5 days. It has the rate of eclosion, 67.4%, in the constant temperature. At the 18 degrees C, H. biruensis has the highest rate of mating and spawning, 92% and 98%, and its rate of mating and spawning varies in the different temperature conditions. The male moth cherishes average 814.4 eggs and lays 607.2 eggs. In the open country, the optimum air relative humidity for mating and laying eggs is 75%-90%, while indoors, the optimum air relative humidity is 78%. PMID- 12776527 TI - [HPLC determination of strychnine and brucine in Semen Strychni and its processed products]. AB - OBJECTIVE: This paper reports a HPLC method for determinition of strychnine and brucine in Semen Strychni and its processed products of Jiangxi method and innovated methed. METHOD: SiO2 was used as the stationary phase, n-hexane dichloromethane-methanol-ammonia(47.5:47.5:5:0.35) as the mobile phase, with detection wavelength of 254 nm. RESULT: The contents of strychnine and brucine in the processed products of Jiangxi are lower. CONCLUSION: This method is accurate, simple and reliable. PMID- 12776526 TI - [Comparative study on internal quality of four cultivars of Chrysanthemum morifolium]. AB - OBJECTIVE: To study the genetic diversity of C. morifolium on the chemical constituents. METHOD: Chemical constituents of four cultivars cultivated with the same conditions were compared in three types of index: chlorogenic acid, flavonoid and volatile oil. RESULT AND CONCLUSION: With different cultivars and processing methods, the contents of chlorogenic acid, flavonoid and volatile oil extracted from C. morifolium vary great extent. PMID- 12776528 TI - [Analysis of expansion degree of seeds of Plantaginis and their processed products]. AB - OBJECTIVE: To explore the affection factors to expansion degree of seeds of Plantaginis caused by different processing methods. METHOD: Determination of expansion degree and analysis by gel chromatography. RESULT: The values of expansion degree were decreased and the structures of polysaccharides were changed for the processed products compared with their crude seeds. CONCLUSION: The polysaccharies in seeds of Plantaginis were degraded after toasted without additive and roasted with salt, which further affected the expansion degree. PMID- 12776529 TI - [Changes of dehydroandrographolide's contents of andrographis tablet in the process of production]. AB - OBJECTIVE: To recognize changes in the contents of ingredients of Andrographis Tablet in the process of production. METHOD: Adopting TLCS, TLC, HPLC to detect effective contents of ingredients which are produced in every stage of process of Andrographis Table's production. RESULT: Handling with the fresh Herba Andrographis according to current pharmacopeoia's technology, it showed that only dehyandrographolide can be detected. It indicated that the main factor that leads to chemical change is the heating process in the process of production. CONCLUSION: Avoiding heating treatment or reducing heating treatment time is the main factor to protect the effective ingredients. PMID- 12776530 TI - [Determination of stilbene derivative in shengfa powders by HPLC]. AB - OBJECTIVE: To develop a new method for the determination of the idiosyncratic component, 2,3,5,4'-tetrahydroxy-stilbene-2-O-beta-D-glycoside in Shengfa Powders. METHOD: A HPLC method was set up, using Hypersil BDS C18 column (5 microns, 4.6 mm x 150 mm), acetonitrile-water(18:82) as mobile phase, with detection at 320 nm. RESULT: The calibration curve was linear in the range of 0.45-2.25 micrograms, r = 0.9998, the average recovery was 99.5%, RSD = 1.2%(n = 6). CONCLUSION: The active constituent 2,3,5,4'-tetrahydroxy-stilbene-2-O-beta-D glycoside in Shengfa Powders can be separated effectively. This method is simple, specific and exact. PMID- 12776531 TI - [Determination of ursolic acid in herba of Verbena officinalis by HPLC]. AB - OBJECTIVE: To establish a HPLC method for determination of ursolic acid in dried aerial part of Verbena officinalis. METHOD: The column used was a Kromasil C18 (4.6 mm x 250 mm) packed with a 5 microns stationary phase. The mobile phase consisted of methanol-sodium phosphate buffer [monobasic sodium phosphate (MW = 119.98) 1.7997 g and phosphoric acid (85%) 1.02 mL, combined and brought the total volume of 1,000 mL with water] (89:11); the mobile phase was maintained at a flow-rate of 0.8 mL per minute; the column was maintained at 40 degrees C; the DAD detector was set at 210 nm. RESULTS: The liner range was 0.251-10.04 micrograms (r = 1.0000). An average recovery of 98.1% (n = 6) was obtained with a RSD of 1.0%. CONCLUSION: The method is simple, accurate and suitable for the qualify control. PMID- 12776532 TI - [Analysis of 1H-NMR fingerprint in stem of Dendrobium loddigesii]. AB - OBJECTIVE: To analyse the 1H-NMR finger-print of the stem of Dendrobium loddigesii. METHOD: Silica gel column chromatography was used to separate the chemical constituents of SCE A of the stem of D. loddigesii. The characteristic signals of the H-NMR finger-print were analysed after determining the structures of the compounds isolated from SCE A. RESULT: 1H-NMR finger-prints of the samples of D. loddigesii collected from different regions showed highly characteristic features and reproducibility. Four compounds predominant in SCE A were isolated and their structures were determined by spectral analysis as 1, 2, 3 and 4, respectively. CONCLUSION: Compound 3 and 4 were isolated from D. loddigesii for the first time. The 1H-NMR finger-print of CGE A of the stem of D. loddigesii showed mainly the characteristic signals of the above four compounds and might be utilized for the original authentication of this plant. PMID- 12776533 TI - [Quinic acid esters from herba of Siphonostegia chinensis]. AB - OBJECTIVE: To study the bioactive constituents, quinic acid esters, from Siphonostegia chinensis. METHOD: The compounds were extracted with solvents, isolated by various column chromatography and identified by spectroscopic methods. RESULT: Three quinic acid esters were isolated and identified as 3,4-di O-caffeoylquinic acid(I), macranthoin F(II), and methyl ester of 3, 4,5-tri-O caffeoylquinic acid (III). CONCLUSION: All compounds were isolated from Siphonostegia for the first time. PMID- 12776534 TI - [Studies on the chemical constituents from herba of Corallodiscus flabellata]. AB - OBJECTIVE: To study the chemical constituents from Corallodiscus flabellata. METHOD: The compounds were isolated with macroporous adsorption resin, silica gel column chromatography and identified on the basis of their physiochemical and spectral data. RESULT: Six compounds were obtained and identified as vanillic acid, 3,4-dihydroxyphenylethyl-8-O-beta-D-glucopyranoside, syringic acid, caffeic acid, isoacteoside, ferulic acid. CONCLUSION: All the compounds were isolated from this plant for the first time. PMID- 12776535 TI - [Experimental study on Rhizoma sparganii and Radices zedoariae treating hepatic fibrosis]. AB - OBJECTIVE: To study the effects of rhizoma sparganii and radices zedoariae on hepatic fibrosis. METHOD: The rat immunohepatic fibrosis model was made by intraperitoneal injection of porcine serum and treated with rhizoma sparganii and radices zedoariae. The ALT, GGT, TP, ALb, A/G, IVC, LN, HA and the pathological change of the liver were observed. RESULT: Rhizoma sparganii and radices zedoariae could increase TP, ALb, A/G, decrease ALT, GGT, IVC, LN, HA and improve the pathological change. CONCLUSION: Rhizoma sparganii and radices zedoariae can protect hepatic cells, alleviate degeneration and necrosis, recover structure and function, and reduce the proliferation of fibrous tissue. PMID- 12776536 TI - [Screening of antidepressant fractions of banxia houpu decoction]. AB - OBJECTIVE: To screen the antidepressant fractions of Banxia Houpu Decoction. METHOD: Behavioral despair models in mice were used to evaluate antidepressant activities and the levels of monoamine neurotransmitters in regions of animal brains were determined with spectrophotofluoremetry method. RESULT: The extract of Banxia Houpu Decoction (AE) and its four fractions were able to elicit time dependent reducing of duration of immobility in the tail suspension test and the forced swimming test in mice. Among these test samples, AE, petrol fraction (AE 1) and water soluble fraction (AE-4) were more potent, but there was no significant difference between them. AE significantly increased 5-HT level in mice striatum and NE level in mice cortex, respectively, and did not affect any monoamine neurotransmitters in other regions of animal brain. AE-1 elevated 5-HT level in the striatum and the NE and DA level in the cortex, chloroform fraction (AE-2) only significantly increased DA level in cortex. CONCLUSION: Banxia Houpu Decoction has antidepressant effect. The active parts are in AE-1 and AE-4. The antidepressant action of Banxia Houpu Decoction is probably by mediated in part through many monoamine neurotransmitter systems. PMID- 12776537 TI - [The inhibiting effect of genistein on the growth of human breast cancer cells in vitro]. AB - OBJECTIVE: To study the inhibitig effects of genistein on the growth of human breast cancer cell lines in vitro and its mechanisms. METHORD: Human breast cancer cell lines both MCF-7(positive estrogen receptor, ER+) and MDA-MB-231 (negative estrogen receptor, ER-) were cultured in vitro. The proliferation of cells was measured with MTT methord and the growth curve was drawn with cell count. The estrogen receptor in cells was show with immunohistochemistry. RESULT: Genistein inhibited proliferation of MCF-7 and MDA-MB-231 cell lines and inhibition was dependent on dose within some concentration range, IC50 being 32.5 mumol.L-1 and 46.8 mumol.L-1 respectively. Genisteins antiproliferation of MCF-7 was stimulated by ectogenesis estrogen but proliferation of MDA-MB-231 inhibited by geinstein was not related to estrogen. The positive signs of ER in cellular nuclei of MCF-7 cell line fed with genistein at concentration of 30 mumol.L-1 were significantly weaker than these not fed with genistein. CONCLUSION: Genistein obviously inhibits proliferation of both cell lines of MCF-7 and MDA-MB 231 in vitro and is dependent on dose. Genisteins antiproliferous effect on MCF-7 cell lines is stimulated by estrogen and this effect is related with ER, but genisteins inhibiting proliferation of MDA-MB-231 cell line is not through ER. PMID- 12776538 TI - [Effects of aqueous extract in herba of Lysimachia christinae on hyperuricemia in mice]. AB - OBJECTIVE: To study hypouricemic effect of aqueous extract of Lysimachia christinae on hyperuricemia in mice. METHOD: The uricase inhibitor potassium oxonate was used to induce hyperuricemia in mice, and serum uric acid level was determined with the phosphotungstic acid method. RESULT: The aqueous extract of Lysimachia christinae, when administered orally to the oxonate-induced hyperuricemic mice at the doses of 5.2, 10.4 and 20.8 g.kg-1, was able to elicit dose-dependent hypouricemic effects. At these doses of the extract, the serum urate levels of the oxonate-pretreated mice showed no difference from the normal mice. In normal mice, however, oral administration of the extract at the same doses did not produce any observable hypouricemic effects. CONCLUSION: The aqueous extract of Lysimachia christinae possesses potent hypuricemic effects on models of hyperuricemia in mice pretreated with oxonate. PMID- 12776539 TI - [Effects of antioxidation and ceramide of improved prescription of didang-tang (IPDT) on exprimental atherosclerosis rabbits]. AB - OBJECTIVE: To observe the effects of antioxidation and ceramide content of improved prescription of Didang-tang (IPDT) on exprimental atherosclerosis(AS) rabbits. METHOD: Plasm Superoxide Dismutase(SOD) activity was detected with micro content fast detecting method, Plasm Malondialdehyde(MDA) content with improved BaMuGuoFu method, and Aortic Ceramide (CER) content with thinlayer scanning. RESULT: IPDT could effectivly improve plasma SOD activity and decrease plasma MDA content and decrease aortic CER content. CONCLUSION: IPDT on exprimental AS is related to the improvement of antioxidation and decrease of CER content. PMID- 12776540 TI - Culture & women's health: building inner strength in women. PMID- 12776541 TI - Development and testing of the inner strength questionnaire. AB - The aim of this research brief is to describe the results of psychometric evaluation of a new instrument to measure inner strength in women with chronic illness. Inner strength is defined as a central human resource that promotes well being and healing (Roux, Dingley, & Bush, 2002). The instrument can be used in clinical settings to examine women's inner strengths associated with cognitive, social, physical, and mental needs to improve health outcomes and enhance well being. The theory of inner strength has important implications for health care providers to improve health outcomes and symptom management for women living with chronic illness. PMID- 12776542 TI - Inner strength in Salvadoran women: a secondary analysis. AB - The purpose of this study was to utilize secondary analysis to identify the impact of challenging life events on the inner strength of women in El Salvador. The women interviewed lived in a rural area repatriated following the civil war that ended in 1992. The original ethnographic study examined health beliefs and practices of these Salvadoran women. The data were collected through face-to-face interviews over a period of one month. The experience of inner strength was evident in the original data, leading to the examination by a secondary analysis. Findings from the secondary analysis revealed the relationship between the women's descriptions of their lifeways and the six attributes of inner strength as described by Dingley, Roux, and Bush (2000). Findings provided further clarification and meaning to the concept of inner strength in women. Implications for practice resulting from this analysis facilitate understanding of behaviors affecting health and illness and provide a rich and holistic understanding of the culture in Salvadoran women. PMID- 12776543 TI - Inner strength in older Hispanic women with chronic illness. AB - The phenomenon of inner strength has been studied in women with various chronic illnesses (Dingley, 1997; Dingley, Bush, & Roux, 2001; Koob, Roux, & Bush, 2002; Roux, Bush, & Dingley, 2001), demonstrating commonalities in the experiences that bridge the boundaries created by disease taxonomy or individual circumstance. Despite the richness of previous qualitative studies, the current state of the science reflects the interpreted reality of inner strength from the perspective of primarily middle class Caucasian women. Therefore, the purpose of this qualitative descriptive study was to explore the experience of inner strength in older Hispanic women living with chronic illness. The primary research question was: How do older Hispanic women experience inner strength as they live with chronic illness? The purposive sample consisted of women who met the criteria for the term "Hispanic", were 60 years of age or older, and had been diagnosed with a chronic illness. Data generation methods included observation and semi-structured interviews. Data analysis was based on qualitative methodology informed by grounded theory and the constant comparative method. Findings from the current study suggested the experience of inner strength in older Hispanic women involved the following interrelated dimensions: (a) Drawing Strength from the Past, (b) Focusing on Possibilities, (c) Being supported by Others, (d) Knowing One's Purpose, and (e) Nurturing the Spirit. The current study contributes to a cumulative program of research on the phenomenon of inner strength, furthering theory generation, cultural awareness, and future intervention studies. PMID- 12776544 TI - Postpartum depression, culture and African-American women. PMID- 12776545 TI - Female genital circumcision: medical and cultural considerations. AB - Female circumcision (FC), also known as female genital mutilation (FGM), is a procedure that involves partial or complete removal of external female genitalia. The definition given by the World Health Organization (WHO) states that female circumcision "comprise all procedures involving partial or total removal of the external female genitalia or other injury to the female genital organs whether for cultural, religious or other non-therapeutic reasons" (WHO, 1998, p.5). The United Nations Children's Fund, the United Nations Population Fund, and the WHO have jointly issued a statement that FC and FGM causes unacceptable harm and issued a call for the elimination of this practice worldwide. The WHO also contends that female circumcision is a "violation of internationally accepted rights" (WHO, p.1). Female circumcision is a widespread cultural practice and affects millions of young women. Issues related to female circumcision that are of special concern are health consequences, civil rights, cultural considerations, and legal and ethical aspects. The purpose of this paper is to address the incidence of FC and FGM, the historical background, the procedure, the medical complications and cultural considerations. Legal and ethical issues of FGM will also be discussed. PMID- 12776547 TI - [Study on the micro-identification of insect material medicine]. AB - OBJECTIVE: To seek microstructure characters and method of insect material medicines. METHOD: Insect material medicines and Chinese patent medicines containing these medicines were observed and compared with OLYMPUS. RESULT: The texture of integumentary cells, the shape and size of hair socket and bristle were important characters for species identification. CONCLUSION: This method can effectively identify 8 insect medicines, which provides some data and foundation for Chinese patent medicines' identification containing insects medicines and making quality standard for new drugs. PMID- 12776546 TI - [A review on the study of hubeibeimu]. AB - Hubeibeimu has been used with a long history. Now as one of the major sources of Beimu, it contains 10 steroid alkaloids, 2 diterpenes and 3 diterpenoid dimers. Has bronchial smooth muscles relaxation, antitussive, expectoant and antiasthmatic activities, with very low toxicity. Modern research is in accordance with the traditional experiences of TCM, and Hubeibeimu has been recorded in the latest Chinese Pharmacopoeia (2000 ed.), which is worthy of further development. PMID- 12776548 TI - [In vitro culture and the Agrobacterium-mediated genetic transformation of Morinda officinalis]. AB - OBJECTIVE: To establish an effective system for the Agrobacterium-mediated genetic transformation of M. officinalis, for laying a foundation for the improvement of breeds and introduction of foreign objective genes. METHOD: The explants used for culture were the nodular stem segments from M. officinalis. Agrobacterium tumefaciens strain was EHA101, containing vector plasmid pGA482GG. The GUS gene and NPT II gene were introduced into the plasmid. RESULT: MT basal medium with BA 1 mg.L-1 was effective to inducing the direct shoot formation, and the frequency of shoot formation was 97.8%. As BA concentrations increased, the ability of shoot formation decreased. The explants oriented with their apical ends protruding from the medium produced more shoots than when they were placed with their basal end upright or were placed horizontally. The optimal rooting medium for regenerating shoots was MT basal medium supplemented with 0.2 to 0.5 mg.L-1 NAA, and a root induction rate over 80.0% was observed. The selection pressure for kanamycin was 50 mg.L-1. Cefotaxime was used as antibiotics, and the concentration was 300 mg.L-1. After 1.5 months, 14.8% resistant shoots were emerged from the explants. Histochemical GUS assay showed that 22.2% of the resistant plants were GUS-positive. CONCLUSION: Plant regeneration system and Agrobacterium-mediated genetic transformation have been established for M. officinalis in vitro. PMID- 12776549 TI - [Stuides on evaluation of Semen Armeniacae amarum]. AB - OBJECTIVE: To systematically evaluate the quality of Semen Armeniacae Amarum. METHOD: The amygdalin content in the samples of Semen Armeniacae Amarum from different markets was analysed by HPLC. Some physical and chemical tests were done to investigate their quality. RESULT: Some significant differences in amygdalin contents and some physical and chemical parameters were shown between the crude drugs and the processed ones. There are few changes in amygdalin contents with the storage in 17 years. CONCLUSION: The processing conditions are the main factors that affect the quality of Semen Armeniacae Amarum. Furthermore, a systematic evaluation of the quality of its samples from the different markets has been established by a radar diagram based on the following five parameters: amygdalin contents, the lipid content, ethanol-soluble extract, total ash and acid-insoluble ash. PMID- 12776550 TI - [Kinetic study on the volatility of menthol and borneol]. AB - OBJECTIVE: To study the volatile regularity of menthol and borneol in granule and lozenge at different temperature. METHOD: Kinetic method with GC being detection technique. RESULT: The volatility of menthol and borneol acts as a pseudo first order reaction in open system. Under the same condition, the volatile rate of menthol and borneol in granule is four times as fast as that in lozenge, and the volatile rate of borneol is faster than that of menthol. CONCLUSION: This study can be applied to improve the quality of lozenges containing menthol or/and borneol. PMID- 12776551 TI - [Studeis on the constituents of alkaloids and saponins of ginseng sini tang]. AB - OBJECTIVE: To study the chemical constituents of Ginseng Sini Tang. METHOD: The constituents were identified by physico-chemical properties and spectral analysis. RESULT: The 12 compounds were identified as ginsenoside-Rb1,-Rb2,-Rb3, Rc,-Rd,-Re,-Rg1,Rg2,Rg3,Rf,Ra1,Ra2. The 10 compounds were identified as benzoylmesaconitine(BM), benzoylaconitine(BA), benzoylhypaconitine(BH), neoline (NL), fuziline (FL), 14-ethyl-talatisamine14-acetyl-talatisamine (AT), 14 benzoylhypaconine-8-linoleate (HAL),14-benzoyldeoxyaconine-8-oleate(HAO), 14 benzoylhypaconine-8-palmitate(HAP), talatisamine(TS). CONCLUSION: All these compounds were obtained from Ginseng Sini Tang for first times. PMID- 12776552 TI - [A study on chemical constituents in the herb of Mentha spicata]. AB - OBJECTIVE: To investigate the constituents of anti-inflammatory and hemostatic active sites of Mentha spicata. METHOD: Chemical constituents were separated by solvent method and chromatography and identified by physicochemical properties and spectroscopic methods. RESULT: The structures were identified as ursane I, 3 methoxy-4-methylbenzaldehyde II, veratric acid III, 5-hydroxy-3',4',6,7 tetramethoxyflavone IV, diosmetin(V), thymonin VI, daucosterol VII. CONCLUSION: Among them, compounds I, II, III were first obtained from M. genus, IV, VI were separated from M. spicta for the first time. PMID- 12776553 TI - [Studies on the chemical constituents in radix Astilbes chinensis]. AB - OBJECTIVE: To isolate and determine the chemical constituents from Astilbe chinensis. METHOD: Four compounds were isolated and purified by extraction and column chromatography on silica gel. The chemical structures were determined on the basis of physical-chemical evidence and extensive spectral analysis(HR-EIMS, EIMS, 13C-NMR, 1H-NMR, DEPT, 1H-1H-COSY, HMQC and HMBC). RESULT: Their structures were elucidated as beta-sitosterol palmitate I, daucosterol II beta-sitosterol III and Bergenin IV, respectively. CONCLUSION: Compound I and II were isolated from A. chinensis for the first time. PMID- 12776555 TI - [Study on HPLC fingerprint of the triterpene acids in Poria cocos]. AB - OBJECTIVE: To establish a method for HPLC fingerprint determination of the triterpene acids in Poria cocos. METHOD: RP-HPLC, linear gradient elution and LC/MS, etc. were used to optimize the fingerprint determination method, and identify the main peaks in the HPLC fingerprint. RESULT: A preferable method for HPLC fingerprint determination of the triterpene acids in P. cocos was established, and 9 peaks in the HPLC fingerprint were identified. CONCLUSION: A general acquaintance of the triterpene acids in P. cocos can be obtained by using the preferable HPLC fingerprint determination method, which is useful for quality evaluation of the crud drug of P. cocos. PMID- 12776554 TI - [Studies on chemical constituents in the leaf of Ligustrum delavayanum]. AB - OBJECTIVE: To study the chemical constituents in the leaf of Ligustrum delavayanum Hariot. METHOD: The constituents were isolated with column chromatographies and the structures were identified by MS, IR, UV and NMR. RESULT: Four compounds were isolated and identified as beta-sitosterol, oleanic acid, 2 alpha-hydroxyursolic acid, and acteoside. CONCLUSION: All the compounds were isolated from the plant for the first time. PMID- 12776556 TI - [Determination of quercetin in the Herba Euphorbiae Humifusae]. AB - OBJECTIVE: To develop an HPLC method for determining quercetin in Herba Euphorbiae Humifusae. METHOD: ODS column was used with acetonitrile: 0.4% phosphoric acid (40:60) as mobile phase. The detection wavelength was at 370 nm and the flow rate was 1.0 mL.min-1. RESULT: The linearity was obtained over the range of 0.0364-0.1274 microgram (r = 0.9999). The average recovery was 100.71% with relative standard deviation of 2.45%. CONCLUSION: This method can be used for quality control of Herba Euphorbiae Humifusae. It is simple, rapid and reliable. PMID- 12776557 TI - [Study on the compatibility of composite herbal medicines of the lingguizhugan decoction]. AB - OBJECTIVE: To study the compatibility of composite herbal medicines of the Ling Gui Zhu Gan Decoction. METHOD: Ethanol extract test solutions of the different combinations were prepared according to the orthogonal layout L16(4(5)). Pharmacologic experiments, such as the time of surviving of mice in shortage of oxygen in regular air pressure, the antagonizing effect on arrhythmia induced by chloroform and diuresis were carried out with the solutions. Variance analysis, canonical correlation and stepwise regression analysis were applied to interrelate the amount of each drug and the pharmacologic data. RESULT: The results confirmed that Fuling and Guizhi are the basis, while Baizhuand Gancao are the adjuvans, which is conformed to the theory of TCM. CONCLUSION: This study provides a significant try for studying the compatibility of composite herbal medicines. PMID- 12776558 TI - [Effects of bu yang huan wu decoction on astrocytes after cerebral ischemia and reperfusion]. AB - OBJECTIVE: To study the effects of Bu Yang Huan Wu Decoction on astrocytes after cerebral ischemia and reperfusion. METHOD: Cerebral ischemia model in gerbils was produced by ligating bilateral common carotid artery. The dynamic expressin of GFAP were determined by immunochemistry after cerebyal ischemia for 15 min followed by reperfusion for 24 hours and 48 hours. RESULT: GFAP positive reactions reached a peak after cerebral ischemia for 15 min followed by reperfusion for 24 hours. Bu Yang Huan Wu Decoction decreased the expression. GFAP positive reactions decreased after cerebral ischemia for 15 min followed by reperfusion for 48 hours, Bu Yang Huan Wu Decoction increased the expression. CONCLUSION: The regulation of Bu Yang Huan Wu Decoction on astrocytes after cerebral ischemia and reperfusion may be related to repairing process after cerebral ischemia. PMID- 12776559 TI - [An experimental study(I) on the inhibition of prostatic hyperplasia with extract of seeds of Brassica alba]. AB - OBJECTIVE: To study the effective fraction of the extract of seeds of Brassica alba, which inhibits experimental mice prostatic hyperplasia. METHOD: An experimental model of prostatic hyperplasia of castrated male mice induced by testosterone propionate was made. Fractions I, II and III were prepared by extracting the seeds of Brassica alba successively with ether, ethanol and water under reflux. Total extract was prepared by extracting the seeds of Brassica alba with 60% ethanol under reflux. The total extract and the three fractions were used to test the activities. RESULT: Total extract, fractions I and II could not only significantly inhibit mice prostatic hyperplasia induced by testosterone propionate and activity of serum acid phosphatase, but also decrease wet weight of preputial glands, while fraction III is inactive. CONCLUSION: Extract from seeds of Brassica alba can significantly inhibit mice prostatic hyperplasia induced by exterior hormone, possessing an activity of anti-androgen. Fractions I and II show an equivalent activity of total extract, which indicate that these fractions contain active components of seeds of Brassica alba which can inhibit prostatic hyperplasia. PMID- 12776560 TI - [Effect of gufuhuotang on hemorheology and lipid metabolism of hormonal necrosis of femoral head]. AB - OBJECTIVE: To probe the of Gufuhuotang effect on hemorheology and lipid metabolism of hormonal necrosis of femoral head. METHOD: 32 male rabbits of New Zealand were divided at random into pathological pattern group, normal control group, low-dosage of Gufuhuotang group and high-dosage group; models of osteocyte necrosis of head of femur were made with intragluteal injection of hydroprednisone acetate; the rabbits were observed weekly and killed 6 weeks later and HE pathological section was made to observe hemorheology index and blood-liquid content, and then analysis and comparison were made. RESULT: Gufuhuotang could obviously decrease total blood low-shear viscosity, plasma viscosity, hemacocrit, serum cholesterol and triglyceride contents; under light microscope empty bone lacunas of femoral head were less, fat cells of medullary cavity were less and relatively small, and bone trabecula was sparse with no break. CONCLUSION: Gufuhuotang has a distinct preventive and curative effect on rabbits' hormonal necrosis of femoral head. PMID- 12776561 TI - [Adjustment effect of Radix Astragalus and Radix Angelicae sinensis on TNF-alpha and bFGF on renal injury induced by ischemia reperfusion in rabbit]. AB - OBJECTIVE: To study the mechanism of Astragalus membeanaceus and Angelica sinensis inrenal ischemia reperfusion injury of rabbits. METHOD: Thirty-four heathy mature Japanese big-ear rabbits were randomized into control group, IR model group, Astragalus Membeanaceus + IR groupand Danggui + IR group. Renal ischemia lasted for 1 hour and then the renal reperfusion lasted for 48 hours. At the end, kidneys were observed with Electron Microscopy and the level of Cr in serum and the level of TNF-alpha and bFGF in kidneys were measured. RESULT: The changes of kidneys in IR model group were significant, but they were slight in Astragalus Membeanaceus + IR group and Danggui + IR group. The level of Cr in serum and the level of TNF-alpha in kidneys of IR model group were higher and yet the level of bFGF decreased obviously. The level of Cr and TNF-alpha in Astragalus Membeanaceus + IR group and Danggui + IR group was lower than that in IR model group and the level of bFGF was higher than that in IR model group. CONCLUSION: The mechanism of Astragalus Membeanaceus and Danggui in remedying renal IR injury may be relative to their regulation of TNF-alpha, bFGF and other cytokines. PMID- 12776562 TI - [The antitumor activity of Diosgenin in vivo and in vitro]. AB - OBJECTIVE: To investigate the antitumor activity of Diosgenin in vivo and in vitro. METHOD: S-180, HepA, U14 and EAC transplant mice were given Diosgenin ig or i.p. everyday for 10 days, from the next day when they were inoculated in axilla. Tumor growth inhibit rates were calculated. Four kinds of cells, MCF, L929, A375-S2 and HeLa, were incubated respectively with Diosgenin in vitro. Tumor growth inhibit rates were also calculated. RESULT: In vivo, both ig and i.p., Diosgenin inhibited S-180, HepA, U 14 mice transplant tumor, the inhibit rates being 30%-50%, but it did not inhibit the EAC mice transplant tumor. In vitro, Diosgenin inhibited L929, HeLa, MCF cell growth, and IC50 were 1.2, 18.2, 19.8 micrograms.mL-1 respectively, but it did not significantly affect A375-S2 cells. CONCLUSION: Diosgenin has an obvious antitumor activity on S-180, HepA, U14 transplant mice in vivo and L929, HeLa, MCF cells in vitro. PMID- 12776563 TI - [Research on the quality of original plants and material medicine of Cortex Paeoniae]. AB - OBJECTIVE: To study the contents of main chemical compounds, such as, paeonol, paeoniflorin and their similar compounds in structure in Paeonia ostii and P. suffruticosa, and to find their quality difference. METHOD: HPLC was used. The contents of paeoniflorin and paeonol in 9 collected samples, 5 from P. ostii and 4 from P. suffruticosa, and 6 herb samples bought from different areas were analyzed. The rough contents of 3 paeonol similar compounds and 2 paeoniflorin similar compounds were investigated. RESULT AND CONCLUSION: 1. There is no obvious difference in the contents of paeonol and paeonoflorin and the constitution of other main compounds between P. ostii and P. suffruticosa. Since they belong to different botanic species, it is suggested that both P. ostii and P. suffruticosa should be recorded in Chinese Pharmacopoeia as the origin of Cortex Paeoniae. 2. The glucosides of paeonol might change to paeonol in the course of the collecting and processing of the herb. Therefore when the contents of paeonol is used as the standard to evaluate the quality of the material and patent medicine, the attention should be paid to the transformation mentioned above. PMID- 12776564 TI - [Fungal detection of American ginseng seeds from Beijing and northeast area in China]. AB - OBJECTIVE: The quantity and genus of fungi on 13 samples of American ginseng seed from Beijing and Northeast area in China were detected. METHOD: Washing and surface sterilization were used to isolate the fungi existed on seed surface and inside of the seed, respectively. RESULT: The genera of the fungi in embryo and shell of the seeds mainly belonged to Fusarium spp., Alternaria spp., Penicilium spp., Cephalosporium sp., Rhizopus spp., and a small quantity of Mucor, Aspergillus, Tritroderma, Doratomyces and so on. The isolated fungal frequencies on the shell and in the embryo were 36%-100% and 40%-100% respectively. CONCLUSION: The fungal frequency of American ginseng seed was higher. There were significant differences among different seed samples, the main fungal genera and isolation frequency in the seed embryo were identical with the results from the seed shell. PMID- 12776565 TI - [A study on the percutaneous penetration of artesunate from various reservoir vehicles]. AB - OBJECTIVE: To determine the main factors which affect the percutaneous penetration of artesunate and provide efficient data for the artesunate transdermal delivery system. METHOD: Transdermal speed constant and accumulative amount of 12 hours were used for the estimations of various reservior vehicles, and the supplement orthodox design was used to study the effect of pH, various proportion of IPA/Water/IPM, and drug concentration. RESULT: Drug concentration and pH were the main factors which affected the percutaneous penetration of artesunate. CONCLUSION: The suitable reservior vehicle can prompt the percutaneous penetration of artesunate, and artesunate TTS will be made with further studies. PMID- 12776566 TI - [Determination of nucleosides in siweilingzhi mixture by HPCE]. AB - OBJECTIVE: To establish a method for determining nucleosides (adenoside and guanoside) in Siweilingzhi Mixture by HPCE. METHOD: Adenoside and guanoside were separated within 25 min using an 20 mmol.L-1 borate buffer with 30 mmol.L-1 SDS and 5% Ethanol (adjusted to pH 10.0 with sodium hydroxide solution), with an operation voltage of 10 kV, temperature of 20 degrees C and a hydrodynamic injection time of 15 s. Seperations were carried out in a fused-silica capillary 75 microns id x 57 cm (effective length 50 cm) with peak detection by direct UV at 254 nm. RESULT: Regression equation of adenoside and that of guanoside were Y = 0.0705 + 0.01707X (r = 0.9995) and Y = 0.0232 + 0.01864X (r = 0.9999) respectively. The average recovery rate was 99.22% (RSD = 3.66%) and 104.3% (RSD = 1.91%) respectively. Nine samples were determined with the method. CONCLUSION: The method is simple, rapid and accurate with good repeatability and it can be used to determine nucleosides. PMID- 12776567 TI - [HPLC determination of two flavonoid compounds in Psoralea corylifolia]. AB - OBJECTIVE: To determine two flavonoid compounds in Psoralea corylifolia L. (PC) simultaneously with HPLC method. METHOD: Bavachin and corylin isolated from PC and purified in our laboratory were used as the reference compounds. The HPLC separation was carried out on an Techsphere ODS column using mobile phase consisting of a mixture of methanol and 20 mmol.L-1 ammonium acetate buffer pH 4.0 (67:33), and the UV detection wavelength was 240 nm. RESULT: Simultaneous determination of bavachin and corylin was achieved. The linear range was 1.25-20 micrograms.mL-1 for both bavachin and corylin. The average recovery of bavachin and corylin was 94.9% and 96.2%, and RSD was 3.1% and 3.6% respectively. CONCLUSION: This is the first report on simultaneous determination of bavachin and corylin in PC with satisfactory accuracy and reproducibility. PMID- 12776568 TI - [Studies on the chemical constituents in herba of Galeobdolon chinense (I)]. AB - OBJECTIVE: To study the chemical constituents in the Galeobdolon chinense. METHOD: The chemical constituents were separated by means of chromatogrophy, and their structures were identified by spectra of FAB-MS, IR, UV, 1H-NMR, 13C-NMR, DEPT, HMQC and HMBC. RESULT: Two compounds were obtained and identified as galeobdoside (I) whose structure was apigenin-7-O-beta-D-(6'-p-hydoxy cinnamoyloxy)-mannoside and verbascoside (II). CONCLUSION: Compound I was a new compound and compound II was obtained from this plant for the first time. PMID- 12776569 TI - [Studies on the chemical constituents of Shiraia bambusicola]. AB - OBJECTIVE: To study the chemical constituents of Shiraia bambusicola. METHOD: Column chromatography with silica gel was employed for the isolation and purification of constituents. The structures were elucidated by means of chemical and spectroscopic data. RESULT: Seven compounds were obtained and identified as hypocrellin A (I), hypocrellin B (II), hypocrellin C (III), hypomycin A (IV), ergosterol (V), ergosterol peroxide (VI) and 1,8-dihydroxy anthraquinone (VII). CONCLUSION: Compounds (IV) (VII) were separated from Shiraia bambusicola for the first time. PMID- 12776570 TI - [Intestinal bacteria metabolism of TSDP and characterization of metabolites in rats]. AB - OBJECTIVE: To study the rat intestinal bacteria metabolism of total saponins of Dioscorea pathaica (TSDP) in vitro, and characterize the metabolites in serum and urine of rats after oral administration of TSDP 900 mg.kg-1. METHOD: TSDP metabolites were detected with thin-layer chromatography (TLC) and combination of electrospray ionization mass spectrometry (ESI-MS) and sequential tandem mass spectrometry (MSn). RESULT: In vitro, TSDP was decomposed easily by rat intestinal bacteria, and metabolites DP-1, DP-2, DP-4, DP-5 and diosgenin (Dio) were observed with prolongation of incubation time by ESI-MS2. In vivo, in the full-scan positive mass spectrum of the rat urine sample, the ion peak at m/z 415 (M-H) and its characteristic fragmentations at m/z 397 and m/z 271 in the MS/MS spectrum were identified with that of metabolite Dio, therefore metabolite Dio was deduced to exist in the rat urine, and metablite Dio was allso detected in the rat serum sample. CONCLUSION: TSDP is decomposed easily by rat intestinal bacteria and metabolite diosgenin is absorbed into blood after oral administration of TSDP. PMID- 12776571 TI - [Studies of bi-derectional modulation effect of kudzuvine root on immunol cells]. AB - OBJECTIVE: To evaluate bi-directional modulation effect of Chinese herbal medicine on immunol cells. METHOD: Two different active portions were isolated from Kudzuvine Root(Radix puerariae), one being the ethanol extraction and another the water extraction. Different concentration of these two different portions was studied by using PMA stimulated lymphocyte or eosinophil initiated chemiluminescence system. RESULT: Water extraction of Kudzuvine Root could enhance chemiluminescence concentration dependently whereas enthanol extraction of Kudzuvine Root inhibited the chemiluminescence significantly. CONCLUSION: The bi-directional regulation effect of Chinese herbal medicine can be found in the same herb, because of its efficacy of different active compounds. PMID- 12776572 TI - [Experimental studies of hypoglycemic action on total flavone of Ampelopsis grossedentata from Guangxi]. AB - OBJECTIVE: To study hypoglycemic action of total flavone (GXTF) of Ampelopsis grossedentata from Guangxi by observing the effects of GXTF on blood glucose levels in many strain animal models. METHOD: The blood glucose levels in many strain animal models were determined after oral administration, with the models of diabetes induced by alloxan, of hyperglycemic mice induced by epinephrine and glucose, and normal mice. RESULT: GXTF had better therapeutical action on diabetes mice induced by alloxan, and could significantly lowered the blood glucose levels of hyperglycemic mice induced by epinephrine and glucose, but had no significant effects on blood glucose levels of normal mice. Acute toxicity test showed that the maximum oral dosage is 26.0 g.kg-1. CONCLUSION: GXTF has better hypoglycemic effect on many strain animal models and toxicity is vary small. PMID- 12776573 TI - [Effect of essential oil of Radix Angelicae Dahuricae on beta-endorphin, ACTH, NO and proopiomelanocortin of pain model rats]. AB - OBJECTIVE: To investigate the effect of essential oil of Radix Aangelicae Dahuricae (EOAD) on beta-endorphin Adrenocorticotropic hormone(ACTH), Nitric oxide (NO) and Proopiomelanocortin (POMC) of pain model rats induced by formaldehyde. METHOD: beta-endorphin and ACTH were detected with radio-immunity, the amount of masculine cell expression of Proopiomelanocortin mRNA in hypothalamus, and brainstem of pain model was detected with in situ hybridization and NO with chemical method. RESULT: EOAD could increase beta-endorphin, NO and the amount of masculine cell expression of Proopiomelanocortin mRNA significantly in hypothalamus and brainstem of rats. CONCLUSION: EOAD which may be central analgesic, is associated with endogenous opiate-like substance such as POMC and beta-endorphin. PMID- 12776574 TI - Cardioprotection with sildenafil, a selective inhibitor of cyclic 3',5' monophosphate-specific phosphodiesterase 5. AB - The effects of sildenafil (Viagra), a specific inhibitor of phosphodiesterase 5, on ischemic myocardium was examined using an isolated rat heart model. Rats were pretreated with sildenafil at doses ranging from 0.001 mg to 0.5 mg/kg body weight. After 60 min, isolated hearts were subjected to ischemia for 30 min followed by 2 h of reperfusion. The results demonstrated that at 0.05 mg/kg (and to some extent at 0.01 mg/kg), sildenafil provided significant cardioprotection as evidenced by improved ventricular recovery, a reduced incidence of ventricular fibrillation and decreased myocardial infarction. At higher doses, it caused a significant increase in the incidence of ventricular fibrillation while at very low doses it had no effect on cardiac function. As expected, sildenafil increased cyclic 3',5'-monophosphate (cGMP) content in the heart. The results demonstrate for the first time that within a narrow dose range, sildenafil can protect the heart from ischemia/reperfusion injury, probably through a cGMP-signaling pathway. PMID- 12776575 TI - Dilazep hydrochloride, an antiplatelet drug, prevents progression of diabetic nephropathy in Otsuka Long-Evans Tokushima fatty rats. AB - Diabetic nephropathy is a leading cause of end-stage renal disease in industrialized countries. Although the mechanisms for the development and progression of diabetic nephropathy are not fully understood, platelet activation may participate in its pathogenesis by promoting microthrombus formation. In this study, we investigated the effects of dilazep hydrochloride, an antiplatelet agent, on the development and progression of diabetic nephropathy in Otsuka Long Evans Tokushima fatty (OLETF) rats, a type 2 diabetes mellitus animal model. Administration of dilazep hydrochloride significantly reduced the increase of urinary protein excretions and N-acetyl-beta-D-glucosaminidase (NAG) activity in OLETF rats. Furthermore, dilazep hydrochloride treatment prevented glomerulosclerosis and tubular atrophy and reduced positive staining for type IV collagen in the glomeruli of diabetic rats. These results indicate that platelet activation plays a dominant role in the development and progression of diabetic nephropathy. Our study suggests that dilazep hydrochloride is a valuable new drug for the treatment of diabetic patients with nephropathy. PMID- 12776576 TI - The effects of acarbose and Rumex patientia on liver ultrastructure in streptozotocin-induced diabetic (type II) rats. AB - The aim of this study was to investigate the effects of acarbose and Rumex patientia on liver ultrastructure in streptozotocin (STZ)-induced diabetic (type II) rats. Forty-two-day-old, neonatal Wistar albino rats were used. They were divided into six groups. STZ was injected into groups 4, 5 and 6 on postnatal day 2. Groups 1 and 5 received water, groups 2 and 6 received 2% decoction of R. patientia grain and groups 3 and 4 received 40 mg acarbose/100 g feed. During the experimental period, blood glucose levels were checked periodically and HbA1c levels were measured from cardiac blood at the end of the experiment. In addition, liver tissue was examined by electron microscopy. Our results showed that glucose and HbA1c levels, which are increased by STZ, were decreased by acarbose and R. patientia. In group 5, most of the mitochondria of hepatocytes were swollen and some hepatocytes contained lipid granules in their cytoplasm. In group 4, no pathological changes were observed in hepatocytes, but some lysosomes were found in their cytoplasms. In group 6, mitochondrial changes were minimal compared with those in group 5, and no lipid granules were observed in hepatocytes. PMID- 12776577 TI - Nanomolar level of resveratrol (trans-3,5,4'-trihydroxystilbene) is required, but is not sufficient, to inhibit the growth of human monocytoid tumor cells through an apoptotic-like mechanism. AB - The chemopreventive activity of resveratrol, a stilbene found in grapes and wine, was evaluated in a human monocytic leukemia cell line at the same concentration (100 nM to 1 microM) as that found in the blood-stream after moderate wine intake. As early as at 4 h after intake, resveratrol exhibited antiproliferative and cytotoxic activity. At the same time, some apoptotic-like phenomena were detected such as cell membrane perturbation (phosphatidylserine-annexin V binding), apolipoprotein (APO)-1/FAS (CD95) expression and mitochondrial (delta psi) depolarization. The anticancer drug camptothecin, used as a positive control, did not significantly increase APO-1/FAS (CD95) levels, while only a modest increase in APO-1/FAS-CD95 ligand (CD95-L) was detected. At 12 h, however, resveratrol at concentrations of 100 nM and 1 microM did not exhibit the same antiproliferative activity and increased cell proliferation was correlated to a significant increase in FAS-L expression. We conclude that treatment with low doses of resveratrol, such as those found after moderate wine intake, is not sufficient to stop human leukemia cell line proliferation and that cell resistance, marked by high FAS-L (CD95-L) expression, could be mediated by low (delta psi) mitochondria-released antiapoptotic factors such as BCL-2. It is also suggested that the synergistic action of other wine components with resveratrol might, at least partially, explain its chemopreventive activity. PMID- 12776578 TI - Inhibition of the histamine wheal by ebastine compared with cetirizine, fexofenadine and loratadine at steady state. AB - The objective of this double-blind, randomized, placebo-controlled, 5-way crossover study was to compare the pharmacodynamic effects of the H1 antihistamine ebastine (10 mg once daily, E10) with those of cetirizine (10 mg once daily, C10), loratadine (10 mg once daily, L10), fexofenadine (60 mg, twice daily, F60 x 2) and placebo (P) after 6 days of treatment in healthy volunteers. The pharmacodynamic variable was the mean percent reduction from baseline (pretreatment) of the wheal area induced by intradermal histamine 0.1% on the morning after 6 days' treatment. A secondary variable was the concentration of histamine required to produce a wheal of area 150 mm2. E10 reduced wheal size more than did P (p < 0.001) or F60 x 2 (p < 0.019). No significant differences were found among E10, C10 and L10. After E10, a significantly greater concentration of histamine was needed to induce a wheal of 150 mm2 than after P (p < 0.001), L10 (p < 0.001) or F60 x 2 (p < 0.001). No significant differences were found between E10 and C10. In conclusion, this study shows that, at the end of the conventional dosing interval, ebastine 10 mg and cetirizine 10 mg once daily in repeated doses suppressed the histamine wheal more effectively than did loratadine 10 mg once daily or fexofenadine 60 mg twice daily. PMID- 12776579 TI - Effect of lecithin on epicutaneous absorption of diclofenac epolamine. AB - The epicutaneous application of nonsteroidal antiinflammatory drugs in localized rheumatic diseases results in a highly targeted antiinflammatory action and is associated with reduced systemic effects. The new diclofenac epolamine (DHEP) salt is much more soluble both in water and in lipid solvent than other diclofenac salts. The pharmaceutical addition of lecithin to DHEP leads to the formation of mixed micelles with high affinity to the cellular component, which guarantees the absorption of the active ingredient. We performed a bioavailability randomized, cross-over study to compare the plasma profiles of diclofenamic acid after repeated epicutaneous administration of the new topical formulation with those of the marketed DHEP formulation without lecithin. Based on a randomization list, 12 healthy volunteers were asked to apply one of the two formulations twice a day for 10 consecutive days. The other formulation was given after a washout period of 1 week. Blood samples were collected before the morning epicutaneous dose on days 1, 3, 5 and 8 of treatment and on day 10 at different sampling times until 24 h after the application. The pharmacokinetic analysis showed a significantly higher plasma concentration of diclofenamic acid after the application of DHEP lecithin, which indicates a better saturation of the subcutaneous tissues underlying the application site. This also indicates increased local availability of the active principle. In conclusion, the new DHEP formulation with lecithin should have a therapeutic advantage compared with the formulation without lecithin, even in cases of short- to medium-term treatments. PMID- 12776580 TI - [The research of estrogen receptor modulator and correlative Chinese herbs]. AB - OBJECTIVE: To review the study of estrogen receptor modulator and correlative Chinese herbs. METHOD: Based on to the documents in the world, the estrogen receptor modulator and correlative Chinese herbs summarized. RESULT AND CONCLUSION: Estrogen receptor modulator is biological compounds in botany. It can exert faint estrogen-like effects by low affinity with estrogen receptor. Some of the Chinese herbs have the estrogen-like activity, but further and more systemic research work are to be done. PMID- 12776581 TI - [Application of pattern recognition to quality assessment of the traditional Chinese medicine]. AB - OBJECTIVE: To review the application of the pattern recognition to quality assessment of the Chinese Traditional Medicine. METHOD: Papers of application of chemical pattern recognition, pattern recognition of stereoscopic features and artificial neural networks to quality assessment of the Chinese Traditional Medicine were collected and the application of different method was reviewed. RESULT: IR, UV, GC, GC-Mass and the sum mass spectra were used to evaluate the chemical character of different samples, and the microscopic features of the epidermis were scored quantitatively by stereology and image analysis on the basis of classical taxonomy,. BP networks were also widely used. CONCLUSION: Chemical pattern recognition, pattern recognition of stereoscopic features and artificial neural networks are of important use in quality assessment of material medica. PMID- 12776582 TI - [The naphtha composing characteristics of geoherbs of Atractylodes lancea]. AB - OBJECTIVE: To find the chemical diversity and characteristics of A. lancea on two levels--individuals and populations, and to discover the chemical essentials for forming geoherbs. METHOD: 47 rhizomes of A. lancea were collected in 7 populations, and 6 naphtha components (1. elemol, 2. hinesol, 3. beta-eudesmol, 4. atractylone, 5. atractylodin, 6. atractylenolid I) in the rhizomes were determined by GC-MS combination. Principal Component Analysis and Cluster Analysis were carried out by SPSS. RESULT: Cluster Analysis of the 6 main components indicated that the chemical components of geoherbs were different from those of the non-geonerbs of A. lancea. Other analysis showed as follows: 1. The general oil of geoberbs were lower than that of non-geoherbs(P < 0.01), but components yielding more than 1% (% of the total oil) were more than non geoherbs(P < 0.01); 2. Hinesol mixing beta-eudesmol was more in non-geoherbs, which atractylodin mixing atractylone was more in geoherbs(P < 0.001); 3. Principal Component Analysis implied that atractylone was the most important component to discriminate geoherbs and non-geoherbs of A. Lancea. CONCLUSION: The naphtha composing characteristics of geoherbs was the special proportionment sale, viz. atractylone: hinesol: beta-eudesmol: atractylodin being(0.70~2.00):(0.04~0.35):(0.09~0.40):1. PMID- 12776583 TI - [Investigation on occurrence of lycium pests and their natural enemies]. AB - OBJECTIVE: To Provide the scientific basis for controlling the lycium pests. METHOD: The kinds and number of pests and their natural enemies were investigated at regular intervals and on the fixed trees. RESULT: There are 13 kinds of lycium pests and 17 kinds of natural enemies, the occurrence of which are closely related. CONCLUSION: Natural enemies can control aphids and psyllidae effectively, but ten-spotted lema and Epithrix sp. must be controlled by pesticides. PMID- 12776584 TI - [Microtuber induction in vitro from Rehmannia glutinosa]. AB - OBJECTIVE: To set up the fittest medium and optimum condition for plantlets to form microtubers. METHOD: Plant hormones, concentration of sucrose, active carbon and light time influenced formation of adventitious root and microtuber in vitro from plantlet of Rehmannia glutinosa (85-5). RESULT AND CONCLUSION: The plantlets were cultivated on the medium of microtuber induction after roots were induced on the 1/2MS medium with IBA 1 mg.L-1. The best medium was MS + 6-BA 2 mg.L-1 + NAA 0.1 mg.L-1 + Sucrose 5%. The fittest incubation temperature was 25 degrees C and the light length was (2,000-3,000 lx) 12 h.d-1. Active carbon and GA3 should not be added to the medium, which was not suitable for the induction of microtuber. PMID- 12776585 TI - [Photosynthetic pigment contents of different germplasm of Rehmannia glutinosa and the relationship between pigments and leaf color]. AB - OBJECTIVE: To evaluate the germplasm of Rehmannia glutinosa on the basis of photosynthetic pigment contents (PPC). METHOD: 20 cultivars were planted on the same condition. On Oct. 23 and Sept. 25, 3 leaves per cultivar were collected on different plants, and 80 mg mesophyll was collected among upper lateral veins and was ground in 96% alcohol, and the supernatant was subjected to measure on a spectrophotometer (Angilent 8453). RESULT: The PPCs among cultivars were significantly different at a P < or = 0.01 level. The results of the measurements were similar. Chlolophyll a was the most abundant pigment, but varied to a great extent among different cultivars. 20 cultivars were divided into 9 homogeneous groups according to the contents of chlorophyll a by Duncan's multiple range test at P < or = 0.05. In addition, the content of chlorophyll a was closely related to leaf color. The cultivars with higher chlolophyll a had deep green leaves, and those with lower had yellow green or pale green leaves. CONCLUSION: PPC was an inherent character and an important index for the germplasm evaluation of R. glutinosa. PMID- 12776586 TI - [Application of beta-cyclodextrin inclusion technique in new dosage form of angelica sinensis essential oil]. AB - OBJECTIVE: To make a solid form of Angelica Sinensis essential oil by preparing its inclusion compound with beta-cyclodextrin and evaluate the inclusion rate, dissolution rate and liberating rate of this complex. METHOD: GC-MS, X-ray diffraction and thin-layer chromatography were applied to characterize the essential oil and the inclusion compound. RESULT: The inclusion compound was shown to form a new phase by X-ray diffraction analysis. GC-MS and TLC check-up corroborated the composition of the free oil and the inclusion oil remaining the same. CONCLUSION: The results showed Angelica Sinensis essential oil was more stable with beta-cyclodextrin inclusion. The preparation of this inclusion compound has advantages in limited amount, long time effect, easily preserved and convenient administration as a new dosage form. PMID- 12776587 TI - [Study on the preparation of venenum bufonis beta-cyclodextrin inclusion complexes]. AB - OBJECTIVE: To study the preparation of Venenum Bufonis beta-cyclodextrin inclusion complexes. METHOD: An optimal condition was established by the uniform design. Under the optimal conditions the Venenum Bufonis beta-cyclodextrin inclusion complexes were prepared with 5 different methods. RESULT: The ball grinding method was superior to other four methods. The bufadienolide inclusion rate of Venenum Bufonis beta-cyclodextrin prepared with ball grinding method was 85.42%. CONCLUSION: Ball grinding method is the best method for the preparation of Venenum Bufonis beta-cyclodextrin inclusion complexes. PMID- 12776588 TI - [Studies on quality control standard of zhishidaozhi tabloid pills]. AB - OBJECTIVE: To develop the quality control standard of Zhishidaozhi Tabloid Pills. METHOD: Applying TLC to identify Radix et Rhizoma Rhei, Fructus Aurantii Immaturus, Rhizoma Coptidis, Radix Scutellariae, and HPLC to determine the content of emodin of Radix et Rhizoma Rhei. RESULT: Radix et Rhizoma Rhei, Fructus Aurantii Immaturus, Rhizoma Coptidis and Radix Scutellariae could be indentified by TLC. Emodin showed a good linear relationship at a rang of 0.0612 0.612 microgram, r = 0.9999. The average recovery was 97.9%, and RSD was 2.1%. CONCLUSION: The methods are accurate and quick, and can be used for the quality control of Zhishidaozhi Tabloid Pills. PMID- 12776589 TI - [Studies on chemical constituents of Sorbaria sorbifolia]. AB - OBJECTIVE: To study the chemical constituents in the medicative part of Sorbaria sorbifolia. METHOD: By means of silica gel re-chromatography and thin layer chromatography, chemical constituents from the CHCl3 and EtOAc extract of the medicative part of S. sorbifolia were studied and their structures were identified by their physical-chemical constants, spectral analysis and comparison with the authorized samples. RESULTS: Eight compounds were obtained and identified as follows: noreugenin (I), wogonin (II), 5,7,3',4'-tetrahydroxy-3 methoxyflavone (III), protocatechuic acid (IV), benzoic acid (V), p-hydroxy benzoic acid (VI), emodin (VII) and daucosterol (VIII). CONCLUSION: All of the compounds were isolated from this genus for the first time. PMID- 12776590 TI - [Studies on chemical constituents in the root of Hedysarum polybotrys]. AB - OBJECTIVE: To study the chemical constituents in the root of Hedysarum polybotrys Hand.-Mazz.. METHOD: The constituents were isolated by Sephadex LH-20 and silica gel column chromatography, and their structures were identified on the basis of spectral analysis. RESULT: Five compounds, medicarpin (I), 3-hydroxy-9 methoxycoumestan (II), 3, 9-dihydroxycoumestan (III), beta-sitosterol (IV), beta sitosterol-3-O-beta-D-glucoside (V) were obtained. CONCLUSION: Compounds II, III, V were obtained from the plant for the first time. PMID- 12776591 TI - [Effect of the purariae-isofiavones on estrogen level in normal and ovariectomized rats]. AB - OBJECTIVE: To study the effect of purariae isoflavone on estrogen level in ovariectomized rats. METHOD: 80 rats were divided into four groups randomly, every group with 20 rats: 1. Control group; 2. Normal + purariae isoflavone group; 3. Ovariectomized group; 4. Ovariectomized + purariae isoflavone group. Estrogen level and gonadotropin-releasing hormone level of all rats were measured. RESULT: Thirty days after being ovariectomized, E2, E3 level was significantly lower than that of the first group(P < 0.05). But Testerone, FSH, LH, PRL and GH increased(P < 0.05). After being gastrogavaged with purariae isolfavone for thirty days, Estrogen level and gonadotropin-relasing hormone level of the second group were significantly lower in various degree than those of normal control group (P < 0.05). But in ovariectomized rats, the estrogen level was recovered (P > 0.05). The gonadotropin-releasing hormone level was increased (P < 0.05). CONCLUSION: Purariae-isoflavone can increase estrogen level to normal in ovariectomized rats by way of increasing the level of gonadotropin releasing hormone. In normal rats, it has anti-estrogen effect. PMID- 12776592 TI - [Depressive effect of total flavonoid fraction of Asttragalus complanatus R. Br and its influence upon hemodynamics in SHR]. AB - OBJECTIVE: To investigate effects of total flavonoid fraction of Asttragalus complanatus on blood pressure in conscious spontaneously hypertensive rats(SHR) and hemodynamics in anesthetized SHR. METHOD: Blood pressure was recorded with tail-culf method and hemodynamics was recorded with polygraph after administration of total flavonoid fraction of Asttragalus complanatus. RESULT: Total flavonoid fraction of Asttragalus complanatus (100, 200 mg.kg-1) could decrease blood pressure of conscious SHR(decreasing 7.1%, P < 0.05 and 9.3%, P < 0.01 respectively). Meanwhile, there was no significant change in heart rate(HR). R Br (200 mg.kg-1) could decrease total peripheral resistance (TPR) (decreasing 20%, P < 0.05) but did not influence the cardiac output(CO) and heart rate in anesthetized SHR. CONCLUSION: Total flavonoid fraction of Asttragalus complanatus possesses obvious hypotensive effect, mainly by means of decreasing the TPR. PMID- 12776593 TI - [Anti-emetic effect of ethanol extract from "Wuzhuyu broth"]. AB - OBJECTIVE: To investigate the anti-emetic effect of ethanol extract from "WuZhuYu broth" and its mechanism. METHOD: Three experiments were carried out test which extract has anti-emetic activity, such as CuSO4-induced pigeon's emetic response, gastric emptying in mice and ACh-induced or 5-HT-induced contraction in vitro gastric muscle in rats. Meanwhile, effect of anti-emetic extract on concentration response curve to ACh, 5-HT, histamine was investigated. RESULT: 50% ethanol extract and 70% ethanol extract were identified as having significantly stronger anti-emetic activities with little side effect, which showed the significant effect on concentration-response curve to ACh, 5-HT, histamine. CONCLUSION: 50% ethanol extract and 70% ethanol extract contain more anti-emetic fractions, more anti-emetic fractions can be gained at the concentrations of 50% and 70% ethanol; the mechanism of anti-emetic effect is related to its antagonism to the receptors of ACh, 5-HT, histamine. PMID- 12776594 TI - [Study on the effect of zhimu combined huangqi on improving renal hypertension rat's cardiac dysfunction]. AB - OBJECTIVE: To observe the effect of Zhimu and Huangqi used singly or combinatively on improving experimental cardiac dysfunction, and mainly to observe zhimu's effect on restraining sympathetic nerve and blocking beta adrenergic-recepter and huangqi's effect of improving hemodynamics on heart failure. METHOD: Two-clib one kidney operation was done to make renal hypertension rat model, 8 weeks after operation, rats were divided into groups and medicated for 6 weeks, and then their heart rate and blood pressure were measured, left ventricle was cannulated to estimate heart function, and heart weight-index and left-ventricle-weight-index were measured. RESULT: Zhimu could slow rats heart rate obviously, prevent cardiac remodeling, but did not affect cardiac function remarkably; Huangqi could reduce blood pressure, heighten +dp/dtmax and -dp/dtmax remarkably; the combined use of the two drugs could decrease plasma catecholamine concentration, adjust myocardium cAMP content, and improve heart function obviously. CONCLUSION: Zhimu and Huangqi can protect experimental cardiac dysfunction, and the combined use is better than the single use, which shows it better to use the two drugs combinatively in treating heart dysfunction. PMID- 12776596 TI - [Studies of promoter methylation status and protein expression of E-cadherin gene in associated progression stages of gastric cancer]. AB - Gastric cancer, like all cancers, is considered to result in part from the accumulation of multiple genetic alterations leading to oncogene overexpression and tumour suppressor loss. More recently, the role of epigenetic change as a distinct and crucial mechanism to silence a variety of methylated tissue-specific and imprinted genes has emerged in many cancer types. The study of DNA methylation changes in gastric cancer has now provided additional clues into the pathogenesis of the disease. E-cadherin as a metastases suppressor is mutationally inactivated in both familial and sporadic forms of gastric cancers. Evidence now suggests that the transcriptional silencing of E-cadherin gene by promotor methylation plays a crucial role in the development and progression of gastric malignancies. In order to further analyze the role of E-cadherin gene promotor methylation in gastric carcinogenesis and progression, we performed the studies of promoter methylation status and protein expression of E-cadherin gene in associated progression stages of gastric cancer. DNA were extracted from the paraffin embedded gastric specimens of dysplasia(23 cases), early cancer (20 cases) and advanced cancer (20 cases). Methylation specific PCR and immunohistochemistry were used to analyze the promoter methylation status and the protein expression level of E-cadherin gene. Our results showed that E-cadherin promoter methylation occurred in all stages of gastric precancerous lesion and carcinogenesis, which suggests E-cadherin promotor methylation is an important event during gastric carcinogenesis and progression. The positive rate of E cadherin promotor methylation in dysplasia, early gastric cancer and advanced gastric cancer was 78.3%, 80% and 90% respectively. There were significant differences between experimental groups and control group(30%), P < 0.05, but no significant differences among experimental groups, P > 0.05. All of advanced gastric cancer examined were completely E-cadherin protein-negative by immunohistochemistry. Fourteen of 20 early gastric cancer were E-cadherin negative. And 23 dysplasia were all E-cadherin-positive. Thirty-one of 34(91%) of the E-cadherin-negative tumours had promotor methylation. This result indicated the downregulation expression of E-cadherin was associated with promotor methylation in early and advanced gastric cancer (P < 0.01). PMID- 12776597 TI - Cloning of hTERT cDNA fragment and application of anti-hTERT monoclonal antibody in mechanism of laryngeal carcinogenesis. AB - The mechanism of telomerase activation is still not clear till now. In order to understand the expressional mode of telomerase and the mechanism of telomerase activation in laryngeal carcinogenesis, we cloned a fragment of hTERT cDNA and prepared a monoclonal antibody against hTERT. We performed immuno-histochemical staining in laryngeal cancer tissues using this antibody. We found that the frequencies of hTERT positive cells were positively correlated with undifferentiation of cancer tissues and that the expression of hTERT was positively correlated with levels of c-Myc, which suggested that c-Myc might play an important role in activation of telomerase. These results revealed that overexpression of c-Myc upregulated telomerase, which in turn resulted in immortalization of laryngeal squamous cells, and this mechanism existed not only in the initiation of laryngeal carcinogenesis but also in the whole process of cancer development. PMID- 12776595 TI - Transforming and secreting expression of human egf in mutant strain WYBS2001 of Bacillus and its functions. AB - Mutant strain WYBS2001 of B. subtilis with strong anti-pathogenic activity was obtained by mutagenic ultraviolet rays. The gene fragment of Human Epidermal Growth Factor(hegf) of 175 bp was synthesized by PCR and the restriction sites Pst I and Hind III, original code and the signal sequence CTTAGA of secreting vector pUS186 were induced in the fragment. The DNA sequencing result revealed that the synthesized fragment was identical with that of human egf. Then the biological engineering strain WYBS2001T with human egf was obtained by transforming pUSE which was constructed by cloning egf into the secreting plasmid pUS186, into mutant strain WYBS2001. The result of RIA showed that hEGF can be found in the supernatant of the cultures and its content was 7.6 ng/ml. And the content can be increased if the proteinase inhibitor was added into the medium. After several generations' culturing, WYBS2001T positive engineering strain can still secrete and express hEGF steadily. The result of experiment showed hEGF had biological activity of proliferation and growth of human cell K562 in vitro. WYBS2001T engineering strain had obvious effect on healing the burned animals' models. This research showed microecological gene-engineering bacteria has good applying foreground. PMID- 12776598 TI - [Isolation and nuclear transfer of ES-like cells colonies derived from embryos being cloning of bovine somatic]. AB - In this experiment, it was designed to carry out proliferous culture of bovine blastocysts(day 7) derived from embryos cloned through bovine somatic cell nuclear transfer, isolating and passaging of ES cells. The cells of blastocysts, which were planted on feeder layer, formed small colonies within 24 h. The nest shape colonies occurred after culturing for 2-3 days. After the colonies in the same shape were isolated and passaged 4-5 times, many different size colonies with monolayer of multi-cells appeared. The colonies that had been passaged 4-5 times were planted into 4-wells multi-dishes without feeder layer. The colonies with monolayer of multi-cells appeared after 24 h, spread all over the bottom of the dishes, emerged epidermis-like cells that appeared reticulate after 4-7 days. These cells were used as donor cells to carry out nuclear transfer. The results showed that 80% (40/50) of the reconstructed embryos cleaved, 5% (2/40) and 2.5% (1/40) of them developed to the morulaes and blastocyst stage, respectively. It revealed that ES-like cells derived embryos constructed through somatic cell nuclear transfer have the developmental potentials. PMID- 12776599 TI - Genetic studies on eight populations of eight locust species from Shanxi Province, China. AB - The genetic structure of eight locust species in three families (Catantopidae, Oedipodidae and Arcypteridae) from Shanxi Province in China was compared using allozyme analysis with horizontal starch gel electrophoresis. Among 17 loci identified in zymograms, Ao-1, Est-3, G3pd-1, Idh-2 and Mdh-2 had low variability with a few alleles. High polymorphism was observed at Ldh-1, Me-1 and Gpi-1. Each of the eight species demonstrated high percentage of polymorphic loci (P = 64.7% 94.1%) but low observed heterozygosity (H0 = 0.024-0.087) due to heterozygote deficiency. It was noted that the migratory locusts usually had higher percentage of polymorphic loci (P = 88.2%-94.1%) than non-migratory species (P = 64.7% 94.1%). The only exception is Oxya chinensis(P = 94.1%). It is reasoned that the higher polymorphism is necessary for migratory species to cope with the environments that might be drastically different from the habitats before migration. The taxon relationships using cluster analysis based on Nei's genetic identity (I) and Roger's genetic distance (D) were the same at species and genus levels. The differences were found at family level, possibly due to the alternative algorithms. The cladogram using Roger's genetic distance (D) overlapped the relationship obtained from karyotypic analyses, which demonstrated that the species examined in Catantopidae displayed somewhat closer relationship to those in Oedipodidae than to those in Arcypteridae. It is suggested that the allozyme analysis is useful as molecular marker for locusts in phylogenetic reconstruction at the species and genus level, while additional data from other studies are necessary when used for higher taxa. PMID- 12776600 TI - [Molecular cloning and sequence analysis of full length cDNA encoding molt inhibiting hormone from Fennropenaeus chinensis]. AB - Molt-inhibiting hormone (MIH) is a neuropeptide member belonging to the eyestalk CHH family. Molting in shrimp is controlled by MIH and ecdysone. By inhibiting the synthesis of ecdysone in the Y-organ, MIH indirectly suppress the molting activity of shrimp. A 697 bp full-length encoding molt-inhibiting hormone precursor cDNA, which has been accepted by GenBank (accession number: AF469187), was firstly amplified from the total RNA of eyestalk from Fennropenaeus chinensis by the 3' and 5' rapid amplification of cDNA ends (RACE) method. The 697 bp full length cDNA encoding MIH precursor was assembled with a 320 bp 3' RACE product and 468 bp 5' RACE product. Results derived from searching by Blast revealed the 697 bp cDNA had high similarity with MIH gene of crustacean. By using Clustal X program, alignment of the amino acid sequence deduced from the 697 bp cDNA with amino acid sequences of 7 MIHs revealed that the deduced amino acid sequence had very high identity with amino acid sequences of MIHs of shrimps. The identities between the deduced amino acid sequence with that of MIH of Marsupenaeus japonicus, Penaeus monodon and Metapenaeus ensis were respectively 95.1%, 83.1% and 79.1%. On the base of all the data, we concluded that the 697 bp full-length cDNA was the cDNA encoding MIH precursor of F. chinensis. Sequence analysis of the 697 bp cDNA revealed a 312 bp open reading frame, and 81 bp 5' untranslated region, and a 302 bp 3' untranslated region. The deduced 103 amino acid polypeptide consisted of a 28 amino acid region of signal peptide and a 75 amino acid region of mature peptide. The six cysteine residues were very conserved in the mature peptide. PMID- 12776601 TI - [Cotransformation of rice by bar and cecropin B gene expression cassettes lacking vector backbone sequences]. AB - Whole plasmids are used in both Agrobacterium-mediated transformation and direct DNA transfer, generally leading to the integration of vector backbone sequences into the host genome along with the transgene(s). The undesirable vector backbone sequences may not only promote transgene rearrangements and affect transgene or endogenous gene expression negatively, but have disadvantage on the safe assessment of the transformants as "desert DNA". The direct DNA transforming systems can transfer minimal gene expression cassettes (promoter, open reading frame, terminator) into plant genome and generate "safer" transformants, also it can delivery multiple genes of agronomic relevance to economically-important crop plants. But there is seldom researching reports on the topic till now. The present paper studied some factors that affecting the transforming efficiency of liner gene expression cassettes to rice varieties by particle bombardment, and the integration patterns of the gene expression cassettes in rice genome were compared with that of the whole plasmids. The results showed: (1) The transforming frequency of gene expression cassettes to rice via particle bombardment is 0.1%-0.5%, the cotransforming frequency of non-selectable gene is about 50%-60% when two separate gene expression cassettes were used for transformation. Increasing the DNA mole content can increase the transforming frequency and the beside sequences of gene constructs may play an important role on the variation of transforming efficiency between different rice varieties. (2) It's reported that the selectable and non-selectable transgene expression cassettes generated low-copy-number transgenic plants with simple integration patterns. While our results showed that the non-selectable cecropin B gene cassette generated simple integration patterns with 1-3 copies in the rice genome, but the selectable bar gene cassette which got 4-14 copies had much more complex integration patterns than that of the whole plasmids which got 1-3 copies only. As the bar gene is promoted by the CaMV35 promoter, in which there is a 19 bp palindromic sequence could act as recombination hot spot and lead to DNA rearrangement, we presumed that the transgene recombination events happened during the integration course have generated the complex Southern patterns of bar gene expression cassette. The recombination character, the heredity behavior and the expression law of gene expression cassettes in the rice genomes will be reported in our future papers. PMID- 12776602 TI - [Effect of temperature on the fertility restoration of different two-line hybrid rice]. AB - On the basis of the fertility observation of hybrids from Pei'ai 64S and Nongken 58S crossed with an indica variety Nanjing11 and japonica marker line FL235, respectively, The plant growth chambers were employed to expose the F2 plant individuals to such different day mean temperature as 24 degrees C, 27 degrees C and 30 degrees C during natural long hour daylight from July to August in Wuhan (30 degrees 27' N), with a view to analyzing the difference in the temperature sensitivity of fertility between the two kinds of different fertility-restoring genes for two years running. The results showed that, whether it was under long daylight and high temperature or under long daylight and middle temperature, the mean natural seed-set percent of F1 was higher than 68.75%, suggesting that Nanjing11 could completely restore the fertility of Pei'ai 64S. And however, under natural high temperature condition, N58S x FL235 F1 could set seed naturally with 21.93%-26.75%, the mean natural seed set percent of F1 was 46.36% 48.38% in natural middle temperature condition, ability of FL235 to restore the fertility of N58S was affected by high temperature. Further analysis proved that temperature could not alter the inheritance mode of F2 but affect the extent of fertility genes expression in Nanjing11. On the other hand, the expression of fertility-genes of FL235 was sensitive to high temperature, whose the putative critical temperature was 27 degrees C, and high temperature influenced not only the genetic interactions but also the segregation modes in F2 generations. PMID- 12776603 TI - [A new method for EST clustering]. AB - We developed an EST (expressed sequence tag) clustering method, ESTClustering, to generate high-quality unique expressed sequence based on large-scale EST sequencing. The method uses consensus sequences to sequence analyze with megablast and assemble each cluster with phrap in clustering process. The clustering strategy can efficiently identify gene family and alternate splicing forms of expressed sequences. It can also reduce the adverse effects caused by sequence errors. The ESTClustering method tends to provide more expressed gene forms comparing with the UniGene clustering method of the National Center for Biotechnology Information. Analysis of the 112,256 ESTs of Arabidopsis with ESTClustering produced 23,581 EST clusters. Among these Arabidopsis EST clusters, 13,597 have corresponding genome coding sequences and this number is close to the number of genes predicted with Arabidopsis ESTs. Using this clustering method, a total of 147,191 rice ESTs were clustered into 33,896 groups. PMID- 12776604 TI - Relationship between differential gene expression patterns in functional leaves of maize inbreds & hybrids at spikelet differentiation stage and heterosis. AB - To understand the molecular mechanism of maize heterosis, differential gene expression patterns in functional leaves between 10 maize inbreds and 38 hybrids at spikelet differentiation stage were analyzed by using cDNA-AFLP. The correlation analysis of various differential gene expression patterns with the performance and heterosis of main maize agronomic traits was carried out. The main results are as follows: (1) There are differential gene expression patterns in quality and quantity between hybrids and their parents. The differential expression patterns in quality include: bands expressed only in one parent, bands expressed only in both parents, bands expressed only in one parent and F1, bands expressed only in F1. (2) At spikelet differentiation stage, there are large variations among different hybrids for the same differentially expressed patterns. In general from mean data, there were 25.22% bands expressed only in F1, 21.46% bands expressed in one parent and F1, 8.27% bands expressed only in both parents and 33.49% bands expressed only in one parent. (3) For bands expressed only in one parent, significant positive correlation was detected with the relationship to the performance of plant height. For bands expressed only in both parents, significant negative correlation was detected with the relationship to the heterosis of ear diameter. For bands expressed only in one parent and F1, significant negative correlations were detected with the relationships to the heterosis of rows per ear and seed weight per ear. However, for bands expressed only in F1, and for bands only in two parents or only in F1, no significant correlation was detected with the relationship to the performance and heterosis of all agronomic traits. PMID- 12776605 TI - [Cloning and expression of maize fructose-6-phosphate, 2-kinase/fructose-2,6 bisphosphatase (mF2KP)]. AB - A 2469 bp cDNA encoding entire fructose-6-phosphate, 2-kinase/fructose-2,6 bisphosphatase (F2KP) was cloned from maize (Zea mays L.cv. Ziyunuo 1) by the methods of RT-PCR and rapid amplification of cDNA ends(RACE), on the basis of AF007582, which was isolated from "Yedan 4". The cDNA was designated as mF2KP and the GenBank accession number is AF334143, which contains a 2226 bp open reading from (ORF), encoding a 741 residue polypeptide. There are some differences between the F2KP genes of two maize varieties. The length of 3' non-coding region of mF2KP is 38 bp shorter than that of AF007582. On the 1592th, 1593th and 1605th positions of mF2KP, there is an additional nucleotide respectively compared with AF007582, which cause a shifted reading frame in a small region. Northern blot showed that the expression of mF2KP were significantly different among maize tissues. The Transcription of mF2KP in stem was lower than those in leaves, kernel leaves and male inflorescence, but much higher than that in immature seeds. PMID- 12776606 TI - [Genetic variation of Citrus calli revealed by the ploidy analyser]. AB - The cell DNA content of forty-eight Citrus calli of different genotype were measured by using the Ploidy Analyser. The results showed that 93.8% out of forty eight Citrus calli had double DNA content except that Ruby. Weizhang and Kinnow had little varied cells and did not show second peak. Moreover, Pineapple, Meiwa kumquat, Changsha, Russ navel, Guoqing No. 4 and Carter had DNA varied cells including triploids and aneuploids based on the DPAC software analyse. Among the forty-eight Citrus calli, Pineapple had the highest of the DNA varied cells, which was up to 18.57%, Anliucheng had the lowest, 4.07%. There existed significant difference in the variation among genotypes by Duncan Analyse. At the same subcultured medium and at the same subcultured period, the effect of cultural duration on the variation of calli was not significant. PMID- 12776607 TI - [Cloning and expression analysis of a LIM-domain protein gene from cotton (Gossypium hirsuturm L.)]. AB - LIM-domain protein plays an important role in various cellular processes, including construction of cytoskeleton, transcription control and signal transduction. Based on cotton fiber EST database and contig analysis, the coding region of a cotton LIM-domain protein gene (GhLIM1) was obtained by RT-PCR from 4DPA (day post anthesis) ovule with fiber. The cloned fragment of 848 bp contains an open reading frame of 570 bp, coding for a polypeptide of 189 amino acids. It was demonstrated that the deduced GhLIM1 protein was highly homologous to the LIM domain protein of sunflower (Helianthus annuus), tobacco (Nicotiana tabacum) and Arabidopsis thaliana. Two intact LIM-domains, with the conserved sequence of a double zinc-finger structure (C-X2-C-X17-19-H-X2-C-X2-C-X2-C-X16-24-C-X2-H), were found in the GhLIM1 protein. RT-PCR and Northern blot analysis showed that GhLIM1 gene expressed in root, shoot tip, hypocotyls, bud, leaf, anther, ovule and fiber (4DPA, 12DPA, 18DPA). However it was preferentially expressed in the shoot tip, fiber and ovule. It was proposed that the express of GhLIM1 gene is related to cotton fiber development. PMID- 12776608 TI - [A and D genome evolution in Gossypium revealed using SSR molecular markers]. AB - Genetic diversity analysis of diploid and allotetraploid cotton species was carried out using SSR molecular markers by selecting representative species having A and (or) D genome in Gossypium. Ten diploid cotton species in A and D genomes had high polymorphism and the molecular cluster was consistent with Gossypium classification previously reported by Fryxell. From molecular level, G. gossypioides belonging to D genome had low similarity matrix compared with other diploid D genome species. Diploid cotton species separately having A or D genome had their high similarity matrix. The research supported that G. gossypioides was the most original cotton species among all D genome species, different genomes in Gossypium had common origin and made evolution separately. To better understand genetic events that accompany allopolyploid formation, we add 2 cultivated allotetraploid cotton species in our research materials. However, the results showed that it was not appropriate to study evolution of A and D genome in Gossypium using cultivated allotetraploid cotton species. In order to solve the question, original arboreum, herbaceum and wild types of allotetraploid cotton species should be adopted. Further evolution research on cotton species based on the transcription level of cotton genome is being carried on. PMID- 12776609 TI - Comparing the base usage frequency between bacteria DNA double strand. AB - During the Evolution, effected by select pressure or nature mutation, the compositions of bacteria genomes is various. And many experiences prove that the genes between leading strand and lagging strand is distinctly in copy, transcription and repair. Some scholars presume that the bases distribution is difference between the two strand, to verify the guess, we using the technology of bioinformatics, compare the base usage between the DNA double strand in 17 species bacteria. The result show: 1. there is same bases usage frequency in coding sequence between leading strand and lagging strand 2. There also same bases usage frequency in first codon, second codon and third codon. It suggest that there is a equilibrium between the two strand by the effect of select pressure and nature mutation. PMID- 12776610 TI - A clone at last? PMID- 12776611 TI - Time for a pause. PMID- 12776612 TI - Calling--and coding--the shots. PMID- 12776613 TI - EMRs. What you need to know. PMID- 12776614 TI - HIPAA update and other computer news. PMID- 12776615 TI - HIV. To tell or not to tell. PMID- 12776616 TI - My very weird lawsuit. PMID- 12776617 TI - Get the best deal on an office lease. PMID- 12776618 TI - Better billing, better collections. PMID- 12776619 TI - FTC to doctors: price fixing won't fly. Interview by Wayne J Guglielmo. PMID- 12776621 TI - Tort reform isn't enough. PMID- 12776620 TI - When an insurer won't pay. PMID- 12776622 TI - When the snowbirds return. PMID- 12776623 TI - Going solo. How four doctors are making it work. PMID- 12776624 TI - Going solo. Start-up basics. PMID- 12776625 TI - Helping patients appeal insurance denials. PMID- 12776626 TI - Get even with a hired gun? PMID- 12776627 TI - MSNJ's new president sets goals for the future. Interview by Patricia A. Costante and Paul J. Hirsch. PMID- 12776628 TI - Medical practice in New Jersey. Reflections on the 1930s from the Journal of the Medical Society of New Jersey. PMID- 12776629 TI - Maternal welfare. Article number one. 1936. PMID- 12776630 TI - Homeland security. In New Jersey it means more than duct tape and plastic. PMID- 12776631 TI - Surgical and anesthesia standards in physicians' offices. Alternative Privileges Rule, N.J.A.C. 13:35-4A.12. PMID- 12776632 TI - The depressed patient in medical practice. Diagnosis, treatment, and referral. PMID- 12776633 TI - Opinion: eliminating health disparities. PMID- 12776634 TI - More help for whistleblowers. PMID- 12776635 TI - Helen Lamb: some insight into her life and times--Part 1. PMID- 12776636 TI - What role should education standards play in the manpower shortage? AB - Any assertions that COA did not respond to concerns raised is not an accurate portrayal of the process and does not acknowledge the significant work the COA and the AANA Education Committee did to write standards that are relevant for nurse anesthesia education today. The COA asserts that information taken from the final draft of the standards presented in 2000 based on "a nonscientific online poll" does more to damage the credibility of COA, AANA, and the profession than it does to establish constructive dialogue. Reporting only a portion of the process used to revise the standards is inaccurate at best. This lack of scientific rigor, as appropriately acknowledged in the "Guest Editorial," offers many problems in subsequent assertions made by the authors. The COA will proceed with the established process for a major revision of the educational standards as last published. The evaluation period for COA Trial Standards continues through November 15, 2003, and COA looks forward to continued, constructive dialogue with the community of interest. PMID- 12776638 TI - Surgical mortality and type of anesthesia provider. AB - Although estimates of anesthesia-related deaths today are as low as 1 in 200,000 to 300,000 cases, questions remain about surgical patients' safety related to types of anesthesia providers. We studied the effect of type of anesthesia provider on mortality rates of Medicare patients undergoing 8 different surgical procedures. Risk-adjusted mortality rates were analyzed for 404,194 inpatients undergoing surgery and having complete, unambiguous Medicare bills for anesthesia. Mortality was compared for anesthesiologists working alone, Certified Registered Nurse Anesthetists (CRNAs) working alone, and anesthesia care teams. Procedure-specific risk-adjustment models were derived using stepwise logistic regression. Predictions were adjusted for institutional and geographic factors. Mortality rates for conditions studied ranged from 0.11% to 1.20%. Observed and predicted values by type of provider were not statistically significantly different. Hospitals without anesthesiologists had results similar to hospitals where anesthesiologists provided or directed anesthesia care. PMID- 12776637 TI - Are we really mentoring our students? PMID- 12776639 TI - Suspected methemoglobinemia following awake intubation: one possible effect of benzocaine topical anesthesia--a case report. AB - Methemoglobinemia is a potential negative side effect associated with the use of benzocaine for topical anesthesia. A healthy patient admitted for an outpatient orthopedic procedure developed the clinical symptomatology of methemoglobinemia after topicalization of the airway with benzocaine. The patient subsequently responded to appropriate treatment and was fit for discharge the same day. Methemoglobinemia should be suspected in cases where decreased pulse oximeter readings develop in patients who have received benzocaine preoperatively. Appropriate steps should be taken to support the airway and oxygenation and monitor the patient while medication is administered to reduce methemoglobin to hemoglobin. PMID- 12776640 TI - Anaphylactoid syndrome of pregnancy: a review of the literature with latest management and outcome data. AB - This article is a review of information on the subject of anaphylactoid syndrome of pregnancy or amniotic fluid embolism, as it is more commonly known. Information was found through a computer search of English articles published from 1958 to 2001. Anaphylactoid syndrome of pregnancy has an 80% to 90% mortality rate. It is difficult to diagnose, and once symptoms occur, there is a rapid deterioration in the patient's condition. It has severe manifestations such as shock and coagulopathy. Identification of the exact mechanism and pathophysiology that trigger the syndrome continue to be investigated. Recognition of the event, swift and aggressive treatment, and education of those who provide care in the obstetrical setting give promise of greater survival potential. Although mortality remains high, there are reports of survivors. PMID- 12776641 TI - Comparison of inhaled isopropyl alcohol and intravenous ondansetron for treatment of postoperative nausea. AB - Postoperative nausea, a common complication in patients receiving general anesthesia, was studied in this randomized investigation to compare the efficacy of 70% inhaled isopropyl alcohol and intravenous ondansetron. For the study, 100 healthy women, ASA physical status I or II, scheduled for outpatient gynecologic laparoscopic procedures randomly received 4 mg of intravenous ondansetron or isopropyl alcohol for the treatment of postoperative nausea. Nausea was measured on arrival to the postanesthesia care unit, at first complaint of nausea, every 5 minutes after initiation of therapy until nausea resolution, and every 15 minutes thereafter using a 0 to 10 verbal numerical rating scale. At 5, 10, and 15 minutes, the median verbal numerical rating scores between the ondansetron and alcohol groups were 6.00 and 3.00, 5.00 and 3.00, and 5.00 and 2.00, respectively (P = .002, .015, and .036, respectively). No statistically significant differences were found at any other time interval. Mean times from initiation of therapy to a 50% reduction in nausea between the ondansetron and alcohol groups were 6.3 minutes and 27.7 minutes, respectively (P = 0.022). Based on this study, it seems postoperative nausea can be resolved quicker using 70% inhaled isopropyl alcohol compared with intravenous ondansetron in women undergoing outpatient gynecologic laparoscopic procedures. PMID- 12776642 TI - Antiemetic prophylaxis: pharmacology and therapeutics. AB - Postoperative nausea and vomiting (PONV), a common problem with complex causes, may result in substantial complications. This Journal course discusses the pathogenesis of PONV and reviews antiemetic pharmacology. Corticosteroids, gastrointestinal prokinetics, neuroleptic butyrophenones, phenothiazines, and serotonin receptor antagonists may be prescribed alone or in combination. State of-the-art anesthetic techniques for prevention of PONV are described. Multimodal therapy with combined low-dose antiemetics affecting multiple receptors is suggested to prevent PONV in high-risk patients such as nonsmokers, females with a previous history of nausea, and patients with high postoperative narcotic requirements. PMID- 12776643 TI - Expansion of nurse anesthesia educational programs: where are the barriers? PMID- 12776644 TI - Vulnerable time periods for attrition during nurse anesthesia education. PMID- 12776645 TI - Acute congestive heart failure after laparoscopic cholecystectomy: a case report. AB - Compared with open procedures, laparoscopic surgery is safe with a low incidence of complications. In rare circumstances, however, intraoperative complications such as acute pulmonary edema have been reported. The patient described herein is a 59-year-old woman with obesity, gastroesophageal reflux disease, and chronic obstructive pulmonary disease who developed acute congestive heart failure (CHF) and cardiomegaly immediately following laparoscopic cholecystectomy. She required emergent reintubation, diuresis, and admission to the intensive care unit for postoperative mechanical ventilation. Potential causes of pulmonary edema associated with laparoscopic surgery (extreme Trendelenburg position, venous carbon dioxide embolism, absorption of crystalloid irrigation fluid, cardiopulmonary disease, adverse drug reactions, negative pressure [postobstructive pulmonary edema]) were considered. A process of exclusion revealed that the hemodynamic changes induced by insufflation with an intra abdominal pressure of 20 mm Hg were the most likely causes of the CHF. Suggestions to prevent occurrence of CHF are tight control of hemodynamics with use of invasive monitoring in high-risk patients and gentle, slow insufflation of the abdomen to an intra-abdominal pressure of 15 mm Hg or less. Intraoperative and/or postoperative CHF should be treated with diuretics, intravenous nitroglycerin, arterial vasodilators, and/or inotropic agents as needed. PMID- 12776646 TI - A comparison of tubocurarine, rocuronium, and cisatracurium in the prevention and reduction of succinylcholine-induced muscle fasciculations. AB - Fasciculations are a common side effect of the use of succinylcholine for tracheal intubation. Many anesthesia care providers prefer to prevent them due to a possible association between fasciculations and increased intracranial and intraocular pressures. The purpose of this study was to compare the effectiveness of tubocurarine, rocuronium, and cisatracurium in the prevention and reduction of succinylcholine-induced muscle fasciculations. The study was a prospective, randomized, double-blind, clinical drug comparison. We randomly assigned 40 subjects to 1 of 4 pretreatment groups. Fasciculations were graded on a 4-point scale. A Kruskal-Wallis analysis of variance, used to analyze data collected from the fasciculation scale, demonstrated there was no statistically significant difference in efficacy between tubocurarine and rocuronium for defasciculation or between cisatracurium and saline for defasciculation. Significant differences were shown between the tubocurarine and cisatracurium groups and between the rocuronium and cisatracurium groups. Rocuronium is equally as efficacious as tubocurarine for defasciculation. Therefore, rocuronium is a valid alternative to tubocurarine for defasciculation. Cisatracurium is inferior to rocuronium and tubocurarine for defasciculation. Therefore, the use of cisatracurium is not recommended for defasciculation. PMID- 12776647 TI - A teaching tool in spinal anesthesia. AB - The purpose of the present study was to develop and evaluate a new model for teaching spinal anesthesia to nurse anesthesia students. The new teaching tool was evaluated to determine if the use of a spinal model allowing visual combined with the haptic sense was more effective than the traditional spinal model using primarily the haptic sense. Specifically, was there a difference in the number of successful passes, the amount of time taken for each spinal attempt, and the number of participants who reached 90% proficiency when comparing the use of both spinal models? Data analysis using a paired t test and Wilcoxon signed rank test revealed that the number of successes, amount of time needed to complete 1 needle pass, and point at which 90% proficiency was reached with the newly designed model were significantly greater than with the traditional model. The present study demonstrated that significant differences exist between participants who used both visual and haptic sense and participants who used primarily the haptic sense when performing spinal anesthesia. Findings of this study will be used to provide information that may be used to change the current curriculum for the training of nurse anesthesia students in spinal anesthetic procedures. PMID- 12776648 TI - Anesthetic implications of necrotizing fasciitis. AB - Necrotizing fasciitis, also known as necrotizing acute soft tissue injury (NASTI), commonly occurs as a consequence of group A streptococcal disease. An estimated 9,400 cases of invasive group A streptococcal disease occur annually in the United States, with 600 cases classified as NASTI. Mortality associated with NASTI is estimated at 20% to 50%. Research indicates that early diagnosis and surgical excision of necrotic tissue is the key to minimalization of morbidity and mortality associated with NASTI. Repeated surgeries typically are required in patients with NASTI, posing unusual anesthetic challenges. This article provides an overview of NASTI and includes preoperative, intraoperative, and postoperative anesthetic considerations. PMID- 12776649 TI - Hand ischemia associated with profound hypotension and radial artery catheterization in a pediatric patient: a case report. AB - Direct arterial blood pressure monitoring via the radial artery is a relatively common method employed during the perioperative period. Complications attributed to radial artery catheterization are extremely rare but can include thrombosis, ischemia, infection, and aneurysm formation at the site of catheter insertion. This report describes an episode of hand ischemia in a pediatric patient. The patient experienced a brief period of hypotension secondary to blood loss. A pallor hand was noticed after the patient had been adequately resuscitated. Appropriate treatment was administered, and the patient was eventually discharged to home without any further complications. PMID- 12776650 TI - The frequency of respiratory failure in patients with morbid obesity undergoing gastric bypass. AB - Morbid obesity is associated with multiple metabolic and mechanical abnormalities that increase morbidity and mortality after major abdominal surgery. It is unclear whether patients undergoing bariatric surgery have increased pulmonary complications postoperatively. We performed a retrospective chart review of 207 patients who underwent elective gastric bypass surgery during a recent 2-year period. Body mass index (BMI = kg/m2) of more than 35 was used to define morbid obesity. The purpose of this study was to determine the frequency of respiratory failure, defined as intubation for 24 hours or more and/or reintubation, in these patients. We also evaluated differences in the frequency of respiratory failure between patients with a BMI of 43 or less and those with a BMI of more than 43. Patients with morbid obesity undergoing elective weight loss surgery had few respiratory or other perioperative complications with our experienced team. The rates of respiratory failure and total postoperative complications were 8% in the group with a BMI of 43 or less and 14% in the group with a BMI of more than 43. Skillful anesthetic care allows patients with significant comorbid conditions to benefit from bariatric surgery with reasonable risk in terms of postoperative complications. PMID- 12776651 TI - An evaluation of the histological effects of intra-articular methadone in the canine model. AB - Methadone hydrochloride is an opiate that has pharmacodynamic and pharmacokinetic properties that suggest it may provide longer analgesia than morphine when administered via the intra-articular route. However, no studies to date have been conducted examining the effects of intra-articular methadone hydrochloride on local tissues. Therefore, the purpose of this study was to determine the histopathologic effects of intra-articular methadone hydrochloride on local tissues in the canine knee. Nine canines, 1 to 4 years old, weighing between 20 kg and 23 kg were used. All canines had their knees randomized to receive either bupivacaine, 0.5% with epinephrine 1:200,000 (4.5 mL), and 5 mg methadone hydrochloride (0.5 mL) for the study knee, or bupivacaine, 0.5% with epinephrine 1:200,000 (4.5 mL), and 0.5 mL normal saline for the control knee. Serum methadone hydrochloride levels were obtained on all canines at 6 and 24 hours. Canines were randomly assigned to 1 of 3 groups to be euthanized at either 24 hours, 14 days, or 28 days. Following euthanization and necropsy, synovial fluid levels and tissue samples were obtained and examined for histopathologic changes. Synovial fluid samples noted a few white blood cells at 24 hours and none at 14 and 28 days. Tissue samples showed no histopathologic changes, and serum concentration levels of methadone hydrochloride were negligible. PMID- 12776652 TI - Cancer: new therapies and new approaches to recurring problems. AB - Caring for patients with cancer presents unique challenges to anesthetists. Chemotherapeutic regimens can cause cardiac, pulmonary, and other complications that will influence the anesthesia provider's care. New surgical techniques, including vertebroplasty, vertebrectomy, radiofrequency ablation of the liver, and sentinel node biopsy, present issues related to the surgical techniques and drugs administered. Recurring problems, including tumors of the airway and cardiac tamponade, continue to present challenges for anesthesia providers. Many patients with cancer who undergo surgery not only have acute pain related to the surgical procedure but also have chronic pain that will influence anesthetic and postoperative pain management. This Journal course discusses new therapies and procedures and approaches to recurring problems in cancer care. PMID- 12776653 TI - [Current problems of psychiatric health care--2002]. AB - The paper is a presentation of the most important and actual psychiatric health care problems in Poland: the financial situation of hospitals, the risks faced by the outpatient psychiatric care, maintaining the legal issues of the psychiatric patients, the situation of forensic psychiatry and legal-psychiatric expertise, post-graduate training, the level of diagnosis and therapy, availability of psychotropic drugs. Attention is paid to the importance of giving the programme of psychiatric health protection a status of evident value and that this programme should become a government (national) programme. PMID- 12776654 TI - [Personality traits in patients with unipolar and bipolar disorder]. AB - AIM: To compare the personality traits as described by H. Eysenck in persons with unipolar and bipolar disorder and healthy persons. METHOD: 60 patients with a diagnosis of uni- and bipolar disorder were studied with the Eysenck Personality Questionnaire (EPQ-R). RESULTS: It was noted that there are significant differences amongst the groups in their neuroticism and extraversion but no differences were noted in the domain of psychoticism. Patients diagnosed as having unipolar disorder have a significantly higher level of neuroticism and a lower level of extraversion than those having bipolar disorder. These domains also differentiate the affective disorder patients from healthy individuals. CONCLUSIONS: The knowledge about the individual characteristic personality traits of patients with affective uni- and bipolar disorder can be important for the diagnostic and therapeutic processes in these disorders, therefore further study on these characteristics is necessary. PMID- 12776655 TI - [Comparison of the personality of anxiety disorder patients and somatization disorder patients before and after psychotherapy]. AB - AIMS: The comparison of personalities of somatization disorder patients and anxiety disorder patients before and after intensive, complex, integrative psychotherapy. METHODS: The 16 PF of R.B. Cattell was used in the study. 200 adults, both sexes, 100 patients in each diagnostic group were studied. Groups were similar in the terms of age, level of education and civil state. Patients performed the test twice, before and after psychotherapy. RESULTS: Before the treatment the groups differed significantly (p < 0.05) as to the ego strength and intelligence--these were lower, and tension--higher in the somatization group. Neuroticism, which was increased in comparison with the statistical norm in each diagnostic group, was higher in the somatization disorder group. After the treatment there was no significant difference between groups and the neuroticism decreased to the statistical norm. CONCLUSIONS: Results point out the relation between neuroticism and functional disorders. PMID- 12776656 TI - [Correlations between personality changes and symptom changes in somatization disorders and anxiety disorders--a comparative study]. AB - AIM: The comparison of the relations between personality changes and main symptom changes observed during complex integrative psychotherapy in 100 somatization patients and 100 anxiety disorder patients. METHOD: Two self-descriptive measures -the 16PF of R.B. Cattell and the Symptom Checklist "O" were used in the study. The second measure chosen for the study was a 14-scale screen for the presence of functional disorder symptoms and 5 scales referring to phobias, general anxiety, somatization and conversions. Questionnaires were fulfilled twice--before and after psychotherapy. RESULTS: It was observed that the main symptoms in the group of anxiety disordered patients decreased with the increase of sociability (r = 0.36 p < 0.001), activity and withstanding difficult situations (r = 0.35, p < 0.001) self-confidence (r = 0.21, p < 0.05) and the reduction of submission (r = 0.36 p < 0.001), however the main symptoms in the somatization disorder group lowered with the growth of the ego strength (r = 0.44, p < 0.001; r = 0.41 p < 0.001) and the reduction of imaginative sensitivity (r = -0.31, p < 0.01). CONCLUSIONS: The results point out to the differences in the connections of the symptoms of functional disorders with the personality and confirm clinical reports on differences between somatization patients and neurotic patients. PMID- 12776657 TI - [Personality characteristics of police candidates]. AB - AIM: The aim of the present study is an attempt to study personality characteristics of police volunteers and to determine the correlation between their age and personality features. METHOD: A number of 106 persons have been investigated, between them 12 women and 94 men in the age from 19 to 35 years. A set of standardised psychological tests has been used as a diagnostic tool, e.g. Eysenck Personality Inventory, MMPI, Raven's Test and Similarities and Vocabulary subscales from the Wechsler Adult Intelligence Scale. The analysis of these correlations has been conducted by the use of the Pearson's correlation ratio. RESULTS AND CONCLUSIONS: In the investigated group the IQ results were within an average range. The analysis of the personality profiles delivered by the MMPI test has proved that it is impossible to determine any concrete personality disturbances in police volunteers. Results achieved in the Eysneck's test eliminate the test as a diagnostic tool useful in the qualification of police volunteers no correlation has been found between age of the volunteers and their personality features. PMID- 12776658 TI - [Scharfetter's EPP (Ego-Psychopathology) questionnaire in Polish setting]. AB - AIM: To estimate the basic indices of reliability, validity and feasibility of the Scharfetter's EPP questionnaire--a clinical tool for assessment of ego disorders according to the author's theoretical model. METHOD: A group of 180 adults of clinically and demographically different characteristics were examined including 78 patients with a diagnosis of schizophrenia, 51 patients with depression in the course of affective disorders according to the ICD-10 criteria and 51 healthy persons as a control group. A Polish version of the EPP questionnaire with a slightly modified method of evaluation of subjects' responses was studied. Psychopathology was evaluated with PANSS and KOSS scales and some aspects of the clinical course were noted. RESULTS: A comparable percentage of positive responses was found between the EPP questionnaire in the original Scharfetter's Swiss group and the group under study. A high factor of internal consistency (Cronbach's alpha) both for the whole scale and its 5 dimensions were found. A relatively high correlation in the expected direction between the EPP and other psychopathology measures, as well as with basic characteristics of the course of the illness was established. Indices of disordered self-experience were significantly higher among patients with schizophrenia than with depression, and higher among both groups of patients then among healthy persons. Cluster analysis of EPP led to similar findings and allowed for distinguishing persons in the study into three clusters consisting mainly of: schizophrenic patients (highest profile), depressed patients (middle profile) and healthy persons (lowest profile). CONCLUSIONS: These preliminary, empirical evaluations of the Scharfetter's EPP questionnaire prove that its Polish version reveals rather satisfactory measures of reliability and validity, and observations suggest, that EPP can be successfully used in the clinical work with patients suffering from schizophrenia. The questionnaire indicates its clinical usefulness and prompts to further studies on the tool itself, as well as on the phenomenon of ego disorder for which it has been constructed. PMID- 12776659 TI - [Psychometric value of CES-D scale for the assessment of depression in Polish population]. AB - AIM: The assessment of usefulness of the CES-D scale in the Polish population was studied. METHOD: 50 patients with depression and 150 healthy men and women selected from the general population were examined. In a group of depressed patients the results of the CES-D scale were compared with the Beck scale. The best CES-D cut-off score was defined. Sensitivity and specificity of the CES-D scale in the relation to clinical diagnosis of depression for healthy men and women and depression patients were calculated. Further factor analysis was done and the results were compared with results previously published for North American populations. RESULTS: High reliability of the CES-D scale (Cronbach's alpha coefficient = 0.92), significant correlation (r = 0.79) and moderate agreement (Kappa Cohen coefficient = 0.44) with the Beck scale was reported. It seems that CES-D cut-off score of 16 is adequate for diagnosis of depression. Sensitivity and specificity of the CES-D scale in the relation to clinical diagnosis of depression were respectively 86% and 77%. CONCLUSIONS: Therefore it is well suitable for detecting depression in the Polish population and its results are similar to those obtained for North American populations. PMID- 12776661 TI - [Self-rating scales in schizophrenia: assessment of the practical applicability of the Paranoid-Depression Scale (PD-S), the Frankfurt Self-feeling Scale (FBS) and of two visual analogue scales]. AB - AIM: To assess the practical feasibility of the Paranoid-Depressivity Scale (PD S, Paranoid-Depresivitats-Skala), the Frankfurt Self-feeling Scale (FBS, Frankfurter Befindlichkeitsskala), and two visual analogy scales of: sense of illness (WAC) and self-feeling (WAS). METHOD: 210 patients with schizophrenia of various clinical courses. All patients in the study group were required to complete each scale twice, at 48 hr intervals. For statistical analysis, two sets of data were singled out (1) the relationship between refusal/inability to complete the scale repeatedly and the selected clinical variables; (2) observations made by the doctor, while the patient was completing the scales. RESULTS: Statistical analysis revealed, that clinical factors like restlessness, autism, maladaptation, recurrence/remission and lower educational status imply a significantly lower readiness of the patient towards completion of self-rating scales. The visual analogy scales were those more readily--and easily--completed by the patients; their interpretation, however, is difficult. Given this, the authors of this paper decided to shorten the questionnaire scales, which reducing the whole to aspects of vital diagnostic relevance. Whether this operation would influence the scales' diagnostic value, required ex-post analysis, which was subsequently performed. Summary analysis revealed that the application of the shortening of the PD-S and FBS had no significant impact on these scales' reliability and validity indices; furthermore, it resulted in a significant decline in the number of ambiguities and thus improved the comprehensiveness of the questionnaire's structure. Also, a significant increase in concordance between the self-rating results and the clinicians' diagnoses was observed. CONCLUSIONS: Visual analogy scales are the more readily and easily applied in practice; a comprehensive interpretation is, however, virtually impossible. On the other hand, the PD-S and FBS are the more difficult for the patients to complete, due to their length, but their interpretation does not constitute a major problem. Ex-post analysis reveals, that shortening the complex questionnaires to their core aspects does not affect their psychometric value negatively; in fact--the contrary. The latter statement, however, requires verification in prospective studies. PMID- 12776660 TI - [Self-rating scales in schizophrenia: validity assessment of the Paranoid Depression Scale (PD-S), the Frankfurt Self-feeling Scale (FBS) and of two visual analogy scales]. AB - AIM: To assess the validity of the Paranoid-Depressivity Scale (PD-S, Paranoid Depresivitats-Skala), the Frankfurt Self-feeling Scale (FBS, Frankfurter Befindlichkeitsskala), and of two visual analogy scales: the Sense of Illness Scale (WAC) and the Self-feeling Scale (WAS). METHOD: 210 patients with schizophrenia of various clinical courses. Diagnostic validity was evaluated by comparing the results of self-rating with clinical assessment using the CGI, KOSS C, BPRS, and KOSS-W scales. Content validity was evaluated by analysis of the results' correlation with given clinical and social-demographic variables. Theoretical (construct) validity was evaluated through factorial analysis with Varimax rotation of the principal elements. RESULTS: The correlation between the self-rating scales and the clinical assessment scales was moderate in the case of questionnaire scales and low for the visual analogy scales. Scales of a similar type were found to correlate to a very high degree. Correlation with clinical assessment depended significantly on the phase and degree of disorder--it was lower in periods of exacerbation and higher during remission. The results of the complex questionnaire scales en somme, correlate better with symptoms considered as typical for schizophrenia, than with others, whereas the isolated self-rating constructs (paranoid, depressive) correlate well with the appropriate dimensions of clinical assessment. There was no significant correlation between the clinical symptoms and dimensions, and the results of self-rating using the two visual analogy scales. Factorial analysis revealed that the internal structure of the basic constructs of the questionnaire scales (general attitude, mood, self feeling) was based on a very rational concept, as well as the fact that the factors isolated have a rational foundation within the theoretical and clinical picture of schizophrenic disorders. CONCLUSIONS: The validity of self-rating scales remains a complex question; analysis of diagnostic, content and theoretical validity seems to confine this method to the role of a complementary, albeit interesting, diagnostic tool; the extent and importance of this role, however, is still to be investigated. PMID- 12776662 TI - [Coping with stress in various rescue groups]. AB - AIM: The main purpose of this study was checking if the above mentioned groups differentiate from each other according to the coping styles used by them; if yes, what does it depend on? METHOD: The Authors studied three groups of rescue services, representing three different formations. There were: Firemen, Ambulance services members and Policemen (preventive troupes). A few well known psychological methods were used, among them: CISS-Coping Inventory for Stressful Situations (in polish adaptation), State-Trait Anxiety Inventory (in polish adaptation) and Eysenck's-Maudsley Personality Inventory (in polish adaptation). RESULTS: The obtained results allowed to formulate some important conclusions. There were no statistically significant differences among the groups as far as the two coping styles were concerned--task and avoidance oriented. Emotionally oriented coping style is the one in which there are statistically significant differences between the researched groups. The Firemen group has the highest score in the emotional coping style. CONCLUSIONS: Using the coping styles (the most effective too), dependence mostly on the personality traits. It means that personality traits should be an essential condition in the way of accepting new members of rescue groups. The personality traits, beside the professional training, are responsible for effective coping in extremely difficult situations. PMID- 12776663 TI - [Use of artificial neural networks in clinical psychology and psychiatry]. AB - Artificial neural networks make a highly specialised tools in data transformation. The human brain has become an inspiration for the makers of artificial neural networks. Although even though artificial neural networks are more frequently used in areas like financial analysis, marketing studies or economical modelling, their application in psychology and medicine has given a lot of promising and fascinating discoveries. It is worth that artificial neurol networks are successfully used in the diagnosis and etiopathogenesis description of various psychiatric disorders such as eating disorders, compulsions, depression or schizophrenia. To sum up, artificial neural networks offer a very promising option of research methodology for modern clinical psychology and psychiatry. The aim of this article is only an illustration of the applications of artificial neural networks in clinical psychology and psychiatry. PMID- 12776664 TI - Everybody's doing it. With the U.S. economy limping along, the unlikeliest marketers are trying to claim a niche in healthcare. AB - The hospital outlook might seem bleak to some investors, but a bevy of diverse companies are seeking the financial cure they believe the healthcare industry can provide. Everyone from carpet companies to trucking firms has been drawn to healthcare because of its seemingly endless consumer demand. Jeannine Rossignol, left, senior marketing manager at Xerox Corp., demonstrates a product at the recent VHA Leadership Conference in Boston. PMID- 12776665 TI - Keep competing. Fla. settles case involving noncompete CON clauses. PMID- 12776666 TI - Exception to the rule. Fla. gives CON break to facility of GOP contributor. PMID- 12776667 TI - Surprise inspection. Bills proposed to enforce Calif.'s nurse-staffing laws. PMID- 12776668 TI - Rural aid. Bill would hike reimbursements to rural hospitals. PMID- 12776669 TI - Full disclosure. Home health quality data go online. PMID- 12776670 TI - To take or not to take. Public N.Y. system under fire for taking vendors' gifts. PMID- 12776671 TI - Post-acute pain. Modern Healthcare survey shows mixed results for providers in '02, with end of Medicare add-on payments among the challenges. PMID- 12776672 TI - Leading the leap. Delbanco, as head of the Leapfrog Group, puts family experience, policy background to good use. PMID- 12776673 TI - The big secret about confidential settlements. PMID- 12776674 TI - Want more young patients? PMID- 12776675 TI - Overworked doctors sue for a breather. PMID- 12776676 TI - Look behind a patient's words. PMID- 12776678 TI - Insurance every practice absolutely must have. PMID- 12776677 TI - Have you been vaccinated? PMID- 12776679 TI - How I got my wake-up call. PMID- 12776680 TI - Document your reasoning, too. PMID- 12776681 TI - Experience or evidence? PMID- 12776682 TI - [History and examination for dentoalveolar surgery]. PMID- 12776683 TI - [Dental extraction: do it yourself or refer?]. PMID- 12776684 TI - [The third molar]. PMID- 12776685 TI - [Apex resection versus endodontic treatment]. PMID- 12776686 TI - [Medication and care with dentoalveolar surgery]. PMID- 12776687 TI - [Legislative aspects of surgical complications]. PMID- 12776688 TI - [Diagnosis and treatment of dentoalveolar trauma]. PMID- 12776689 TI - [Geriatrics and DRG: status at the beginning of 2003]. PMID- 12776690 TI - HT or HRT, that is the question? PMID- 12776691 TI - [Comment on the committee initiative to change the case cost regulation]. PMID- 12776692 TI - [March 2003 revision of the standardized graduate education regulation]. PMID- 12776693 TI - Determinants of body mass index in women around menopause attending menopause clinics in Italy. AB - OBJECTIVE: To analyze determinants of body mass index (BMI, kg/m2) in women attending menopause clinics in Italy. METHODS: Eligible for the study were women attending a network of first-level outpatient menopause clinics in Italy for general counselling about the menopause or treatment of menopausal symptoms. Women observed consecutively during the study period were eligible. A total of 49 122 women (mean age 54 years) entered the study. RESULTS AND CONCLUSIONS: The mean BMI increased slightly with age, being 25.8 (standard deviation, SD 4.8) in women aged < 50 years and 26.3 (SD 4.6) in those aged 57 years or more. This trend was statistically significant also after taking into account the potential confounding effect of menopausal status. The mean BMI was higher in less educated women (27.2, SD 5.1) than in those with high-school education or a university degree (25.0, SD 4.5) (p < 0.05), in non-smokers (26.4, SD 4.9) than in smokers (25.4, SD 4.5), in never-users of hormone replacement therapy (HRT) (26.3, SD 4.9) than in ever-users of HRT (25.4, SD 4.4) and in women self-reporting no physical activity (26.5, SD 4.9) than in those reporting regular physical activity (24.9, SD 4.2). The BMI was higher in women following a surgical menopause than if it was spontaneous (p < 0.05), but there was no difference between the mean BMIs of premenopausal women and those with a spontaneous menopause. Women with diabetes and hypertension had a higher BMI. There was no relation between history of osteoporosis/osteopenia and BMI. PMID- 12776694 TI - [BAG Meeting 25 February 2003 in Hofgeismar]. PMID- 12776695 TI - [Lesson from evidence-based medicine: the need to control all cardiovascular risk factors in treatment of diabetes mellitus]. PMID- 12776696 TI - [Evidence-based medicine and prevention of osteoporotic disease]. PMID- 12776697 TI - Acute stress shortens the time to onset of experimental allergic encephalomyelitis in SJL/J mice. PMID- 12776698 TI - Meta-analysis finds angioplasty more effective than thrombolytic therapy in treating heart attack. PMID- 12776700 TI - Diastolic dysfunction is common and worrisome, study shows. PMID- 12776699 TI - Diuretics are better first-line antihypertensive therapy than calcium channel blockers and ACE inhibitors. PMID- 12776701 TI - Impact of socioeconomic status in etiology and management of urinary stone disease. AB - INTRODUCTION: The diverse manifestations of urolithiasis provide a very interesting epidemiological study from the standpoints of geography, socioeconomic status, nutrition and culture. Stone disease not only affects the patient, but also the national economy, as the disease is prevalent in the productive age group. There has been a continuous search for the cost effectiveness of different treatment modalities not only to treat the patient but also to prevent its recurrence. MATERIALS AND METHODS: The various socioeconomic and dietary factors playing roles in the etiology of urinary calculi in the stone prevalent areas of the world were analyzed. The impact of urinary tract stone disease on the socioeconomic infrastructure of the patient and national economy was studied. A cost-effectiveness analysis vis-a-vis the safety and efficacy of various treatment modalities in both developed and developing countries was done. RESULTS: The standard of living and level of nutrition have directly influenced the process of urolithiasis amongst the communities of the world. A low socioeconomic status has been linked to endemic bladder calculi seen in many poor countries with people subsisting on a deficient diet. The incidence of upper urinary tract calculi increases with prosperity and more nutritious diets. Replacement of open surgery with minimally invasive techniques (ESWL, PCNL, URS) for treating stones in the urinary tract has greatly reduced the patients' morbidity and mortality and the period of hospitalization and convalescence. This change in treatment has also led to less days of absence from work and could minimize the loss to national economy. Minimally invasive surgery is more cost effective in developed countries because of the short hospital stay. However, in developing countries open surgery still appears to be cost-effective in certain subset of the patients. CONCLUSIONS: There is a need for proper health care and a medical insurance system to take care of the financial burden, especially in developing countries, of a disease with a high magnitude of recurrence and morbidity. The need and type of medical treatment should be appraised continually to accommodate advances in techniques of stone removal. Lastly, the quest for etiology, cost-effective treatment and prevention still continues and even today we cannot stay better than Frere Jacques, 'I have removed the stone, it is up to God to cure the patient'. PMID- 12776702 TI - Government task force suggests counseling for patients with diet-related diseases. PMID- 12776703 TI - [ISQua - The International Society for Quality in Health Care, ALPHA. Principles and standards for health care accreditation]. PMID- 12776704 TI - Six-year follow-up of preventive interventions for children of divorce: a randomized controlled trial. PMID- 12776705 TI - Effect of iron supplementation on incidence of infectious illness in children: systematic review. PMID- 12776706 TI - Prevalence of working smoke alarms in local authority inner city housing: randomised controlled trial. PMID- 12776708 TI - Bibliography. Current world literature. Pharmaceuticals aspects, techniques and devices. PMID- 12776707 TI - Bibliography. Current world literature. Anabolic and catabolic signals. PMID- 12776709 TI - My brilliant career--nurse management. Private investigation. Interview by Nick Edwards. PMID- 12776710 TI - Emergency department overcrowding in Massachusetts: making room in our hospitals. PMID- 12776711 TI - Medical error reporting: professional tensions between confidentiality & liability. PMID- 12776712 TI - Children's mental health in the Commonwealth. PMID- 12776713 TI - MassHealth: dispelling myths and preserving progress. PMID- 12776714 TI - The uncompensated care pool: saving the safety net. PMID- 12776715 TI - Keeping elders home: new lessons learned about supporting frail elders in our communities. PMID- 12776716 TI - Delivering the city's children: municipal programs and midwifery in Los Angeles. AB - In 1915, at the height of a movement in the United States to regulate midwifery, health officials in the city of Los Angeles devised an unusual plan for doing so: they put the city itself in the midwifery business. At the same time that public health officials in Los Angeles enacted traditional regulatory legislation to deal with the "midwife problem," they also established a Division of Obstetrics within the city's health department to provide prenatal and postnatal care for the poor. Unlike the maternity dispensaries of other municipalities, Los Angeles provided physicians to attend home-births. Thus, rather than trying to move the delivery room out of the home, Los Angeles moved physicians in. PMID- 12776717 TI - Ibn Zuhr (Avenzoar)'s description of verrucous malignancy of the colon (with an English translation from Arabic and notes on its Hebrew and Latin versions). AB - Ibn Zuhr (ca. 1091-1161) is the most important physician of Muslim Spain. His Kitab al-Taysir, translated into Hebrew and Latin, is principally a manual of therapeutics containing descriptions of clinical cases. His description "On Verrucae (tha'alil) that Occur in the Stomach [sic]," which deals with an emaciated Berber who evacuates a tumour the size of an apple in his stools, represents the first detailed report of a cancer of the colon. Latin and Hebrew translations of this report are close to the Arabic, but it is doubtful that the Latin text is based on the Hebrew. The possibility of an intermediate Judeo Arabic text should be considered. PMID- 12776718 TI - Muir-Torre syndrome and early detection of internal malignancy. PMID- 12776719 TI - Medical history and the European Union: Papanicolaou and Asklepios. PMID- 12776720 TI - [Abulcasis, Avicenna, and Galen: a forensic investigation by a 14th century Jewish physician]. AB - Through a forensic investigation conducted by a Jewish doctor at the end of the 14th century, this paper aims to determine the quality of the training as well as the ensuing practical knowledge that the doctor possessed. Based on the "authorities" (auctoritates) quoted in his investigation, it appears that the doctor, who likely did not attend medical school, acquired the theoretical knowledge that was taught in such institutions and that was required to obtain a medical licence in Provence. PMID- 12776721 TI - [Toxicity of fenfluramine to the central nervous system]. PMID- 12776722 TI - Low-molecular-weight heparins and heparinoids. PMID- 12776723 TI - Religion, spirituality and health. PMID- 12776724 TI - The decline in bulk-billing and increase in out-of-pocket costs for general practice consultations in rural areas of Australia, 1995-2001. PMID- 12776725 TI - The decline in bulk-billing and increase in out-of-pocket costs for general practice consultations in rural areas of Australia, 1995-2001. PMID- 12776726 TI - Peptides that regulate food intake: glucagon-like peptide 1-(7-36) amide acts at lateral and medial hypothalamic sites to suppress feeding in rats. AB - Glucagon-like peptide 1-(7-36) amide (GLP-1) potently inhibits rat feeding behavior after central administration. Because third ventricular injection of GLP 1 appeared to be less effective than lateral ventricular injection, we have reexamined this issue. In addition, we attempted to identify brain regions other than the paraventricular nucleus of the hypothalamus that are sensitive toward GLP-1-induced feeding suppression. Finally, we examined the local role of endogenous GLP-1 by specific GLP-1 receptor blockade. After lateral ventricular injection, GLP-1 significantly inhibited food intake of 24-h-fasted rats in a dose-dependent fashion with a minimal effective dose of 1 microg. After third ventricular injection, GLP-1 (1 microg) was similarly effective in suppressing food intake, which extends previous findings. Intracerebral microinjections of GLP-1 significantly suppressed food intake in the lateral (LH), dorsomedial (DMH), and ventromedial hypothalamus (VMH), but not in the medial nucleus of the amygdala. The minimal effective dose of GLP-1 was 0.3 microg at LH sites and 1 microg at DMH or VMH sites. LH microinjections of exendin-(9-39) amide, a GLP-1 receptor antagonist, at 1 or 2.5 microg did not alter feeding behavior in 24-h fasted rats. In satiated animals, however, a single LH injection of 1 microg exendin-(9-39) amide significantly augmented food intake, but only during the first 20 min (0.6 vs. 0.1 g). With three repeated injections of 2.5 microg exendin-(9-39) amide every 20 min, 1-h food intake was significantly increased by 300%. These data strongly support and extend the concept of GLP-1 as a physiological regulator of food intake in the hypothalamus. PMID- 12776727 TI - Professions and intestinal infections in 15th-century Florence. AB - The aims behind this study are to verify a possible relationship between intestinal infectious diseases and certain professional activities carried out in 15th-century Florence by some categories of lesser labourers. Sources of the present study were the Libri dei Morti by the Florentine Ufficio della Grascia concerning the years 1424-25 and 1430 characterized by a plague epidemic. In this period, thanks to the particular social and economic circumstances of the times, the Libri dei Morti were enriched with additional notes about the illness that caused the deaths. The tables show an analysis of the data concerning the types of illness, the residence of the deceased and his/her social state. The present study suggests that certain professions were more exposed to infectious pathologies. PMID- 12776728 TI - Negotiating with Dharma Pinnu: towards a social history of smallpox in colonial Orissa. AB - This paper explores the social complexities associated with the history of smallpox in colonial Orissa in Eastern India. It focuses on tribal communities while taking into account their interactions with non-tribal communities and colonialism. This research grapples with the phenomenon of of the Hinduisation of tribes and the way this implies an acceptance as well as a rejection of the various aspects of non-tribal, Hindu society. Thus, I examine the specific elements of non-tribal groups inoculation, black magic, and subversive cults, demonstrating the close affinities some of these features have with tribal communities. PMID- 12776729 TI - The influenza and other epidemics. Med J Aust 1919;1:71-72. PMID- 12776730 TI - Access block: problems and progress. PMID- 12776731 TI - Oral care update: from prevention to treatment. PMID- 12776732 TI - Fatal envenomation by jellyfish causing Irukandji syndrome. PMID- 12776733 TI - Chemical-biological-radiological (CBR) response: a template for hospital emergency departments. PMID- 12776734 TI - Itching bites may limit Ross River virus infection. PMID- 12776735 TI - Measuring outcomes in patients with depression or anxiety: an essential part of clinical practice. PMID- 12776736 TI - Developmentally regulated usage of Physarum DNA replication origins. AB - To determine the extent to which eukaryotic replication origins are developmentally regulated in transcriptionally competent cells, we compared origin use in untreated growing amoebae and plasmodia of Physarum polycephalum. At loci that contain genes transcribed in both developmental stages, such as the ribosomal RNA genes and two unlinked actin genes, we show that there is a similar replicational organization, with the same origins used with comparable efficiencies, as shown by two-dimensional agarose-gel electrophoresis. By contrast, we found cell-type-specific replication patterns for the homologous, unlinked profilin A (proA) and profilin P (proP) genes. proA is replicated from a promoter-proximal origin in amoebae, in which it is highly expressed, and is replicated passively in the plasmodium, in which it is not expressed. Conversely, proP is replicated passively and is not expressed in amoebae, but coincides with an efficient origin when highly expressed in the plasmodium. Our results show a reprogramming of S phase that is linked to the reprogramming of transcription during Physarum cell differentiation. This is achieved by the use of two classes of promoter-associated replication origins: those that are constitutively active and those that are developmentally regulated. This suggests that replication origins, like genes, are under epigenetic control associated with cellular differentiation. PMID- 12776737 TI - Stimulation of c-MYC transcriptional activity and acetylation by recruitment of the cofactor CBP. AB - The c-MYC oncoprotein regulates various aspects of cell behaviour by modulating gene expression. Here, we report the identification of the cAMP-response-element binding protein (CBP) as a novel c-MYC binding partner. The two proteins interact both in vitro and in cells, and CBP binds to the carboxy-terminal region of c MYC. Importantly, CBP, as well as p300, is associated with E-box-containing promoter regions of genes that are regulated by c-MYC. Furthermore, c-MYC and CBP/p300 function synergistically in the activation of reporter-gene constructs. Thus, CBP and p300 function as positive cofactors for c-MYC. In addition, c-MYC is acetylated in cells. This modification does not require MYC box II, suggesting that it is independent of TRRAP complexes. Instead, CBP acetylates c-MYC in vitro, and co-expression of CBP with c-MYC stimulates in vivo acetylation. Functionally, this results in a decrease in ubiquitination and stabilization of c MYC proteins. Thus, CBP and p300 are novel functional binding partners of c-MYC. PMID- 12776738 TI - Covariations in the nuclear chloroplast transcriptome reveal a regulatory master switch. AB - The evolution of the endosymbiotic progenitor into the chloroplast organelle was associated with the transfer of numerous chloroplast genes into the nucleus. Hence, inter-organellar signalling, and the co-ordinated expression of sets of nuclear genes, was set up to control the metabolic and developmental status of the chloroplast. Here, we show by the differential-expression analysis of 3,292 genes, that most of the 35 environmental and genetic conditions tested, including plastid signalling mutations, elicit only three main classes of response from the nuclear chloroplast transcriptome. Two classes, probably involving GUN (genomes uncoupled)-type plastid signalling, are characterized by alterations, in opposite directions, in the expression of largely overlapping sets of genes. PMID- 12776741 TI - Problems with stroke genetics studies. PMID- 12776739 TI - A crucial role for profilin-actin in the intracellular motility of Listeria monocytogenes. AB - We have examined the effect of covalently crosslinked profilin-actin (PxA), which closely matches the biochemical properties of ordinary profilin-actin and interferes with actin polymerization in vitro and in vivo, on Listeria monocytogenes motility. PxA caused a marked reduction in bacterial motility, which was accompanied by the detachment of bacterial tails. The effect of PxA was dependent on its binding to proline-rich sequences, as shown by the inability of PH133SxA, which cannot interact with such sequences, to impair Listeria motility. PxA did not alter the motility of a Listeria mutant that is unable to recruit Ena (Enabled)/VASP (vasodilator-stimulated phosphoprotein) proteins and profilin to its surface. Finally, PxA did not block the initiation of actin-tail formation, indicating that profilin-actin is only required for the elongation of actin filaments at the bacterial surface. Our findings provide further evidence that profilin-actin is important for actin-based processes, and show that it has a key function in Listeria motility. PMID- 12776740 TI - Widespread PrPSc accumulation in muscles of hamsters orally infected with scrapie. AB - Scrapie, bovine spongiform encephalopathy and chronic wasting disease are orally communicable, transmissible spongiform encephalopathies (TSEs). As zoonotic transmissions of TSE agents may pose a risk to human health, the identification of reservoirs for infectivity in animal tissues and their exclusion from human consumption has become a matter of great importance for consumer protection. In this study, a variety of muscles from hamsters that were orally challenged with scrapie was screened for the presence of a molecular marker for TSE infection, PrPSc (the pathological isoform of the prion protein PrP). Sensitive western blotting revealed consistent PrPSc accumulation in skeletal muscles from forelimb and hindlimb, head, back and shoulder, and in tongue. Previously, our animal model has provided substantial baseline information about the peripheral routing of infection in naturally occurring and orally acquired ruminant TSEs. Therefore, the findings described here highlight further the necessity to investigate thoroughly whether muscles of TSE-infected sheep, cattle, elk and deer contain infectious agents. PMID- 12776742 TI - Recruiting pedigrees in an ongoing North American stroke genomics study. PMID- 12776743 TI - C. David Marsden and Rolex sign. PMID- 12776744 TI - [Avoidance of allergens, immunotherapy or stepped drug treatment. What really helps your allergic patient?]. PMID- 12776745 TI - Dynamics of metacognitive judgments: pre- and postretrieval mechanisms. AB - Two experiments examined the temporal course of pre- and postretrieval mechanisms using a new kind of dynamic metacognitive judgment. Experiment 1 presented participants with primed and unprimed triples of remote associates to a target word and required them to provide repeated metacognitive judgments, 4 times during a 12-s interval, about the likelihood that they would later recognize the target. Both familiarity with the words and the processing time were associated with changes in metacognitive evaluations. Experiment 2 placed pre- and postretrieval mechanisms in opposition by transforming an element of a previously primed question. For transformed questions this led to high initial ratings, which decreased over time, while for novel questions the ratings were initially lower and increased with time. The results are discussed in terms of pre- and postretrieval mechanisms interacting over time. PMID- 12776746 TI - The cost of remembering to remember in event-based prospective memory: investigating the capacity demands of delayed intention performance. AB - Prospective memory tasks are often accomplished during the performance of other activities. Despite the dual-task nature of prospective memory, little attention has been paid to how successful prospective memory performance affects ongoing activities. In the first 2 experiments, participants performing an embedded prospective memory task had longer response times on nonprospective memory target trials of a lexical decision task than participants performing the lexical decision task alone. In the prospective memory groups, longer lexical decision response times were associated with better prospective memory performance (Experiments 1, 2, and 3), a pattern not demonstrated with an embedded retrospective memory task (Experiment 2). The results of Experiment 3 suggest that the retrieval of a delayed intention, or the prospective component, can require capacity. PMID- 12776748 TI - Practice effects on two memory retrievals from a single cue. AB - The effects of practice on 2 retrievals from a single cue were investigated. In Experiment 1, participants were given extended single-task practice and were then tested on a dual memory retrieval task. Perfomance was consistent with a sequential retrieval model proposed by T. C. Rickard and H. Pashler (2003). In Experiment 2, participants practiced both single- and dual-retrieval tasks extensively. Initially, data from all participants indicated sequential retrieval. However, participants who grouped the 2 response outputs were eventually able to perform the dual task with a latency that approached the prediction of a parallel race model. Models that assume a transition from sequential to parallel retrieval with practice, along with other models that assume an immutable retrieval bottleneck at all practice levels, are considered. PMID- 12776747 TI - Differential effects of cue changes and task changes on task-set selection costs. AB - A task-switching paradigm with a 2:1 mapping between cues and tasks was used to separate cue-switching processes (indexed through pure cue-switch costs) from actual task-switching processes (indexed through additional costs in case of cue and task changes). A large portion of total switch costs was due to cue changes (Experiments 1 and 2), and cue-switch costs but not task-switch costs were sensitive to effects of practice (Experiment 1) and preparation (Experiment 2). In contrast, task-switch costs were particularly sensitive to response-priming effects (Experiments 1 and 2) and task-set inhibition (Experiment 3). Results suggest two processing stages relevant during task-set selection: cue-driven retrieval of task rules from long-term memory and the automatic application of rules to a particular stimulus situation. PMID- 12776749 TI - The role of retrieval practice in directed forgetting. AB - Three experiments tested the possibility that retrieval-induced forgetting is responsible for directed forgetting with the list method. In Experiments 1 and 2, additional List 2 retrieval practice was given to determine whether this would increase directed forgetting. In Experiment 1 all items came from a single category, and in Experiment 2 unrelated words were used. In Experiment 3 additional List 2 study accompanied List 2 retrieval practice. There was no evidence that List 2 retrieval practice, with or without additional List 2 study, affected the magnitude of directed forgetting. It was argued that retrieval induced forgetting could not account for these results. Accounts with greater viability include retrieval strategy disruption and a modified version of the dissociation hypothesis. PMID- 12776750 TI - Human navigation in nested environments. AB - Navigation in humans and many other animals relies on spatial representations of their environments. Three experiments examined how humans maintain sense of orientation between nested environments. Subjects can acquire new spatial representations easily without integrating them into their existing spatial knowledge system. While navigating between nested environments, subjects seemed to constantly switch between the currently processed environment by reorienting to approaching environments and losing track of old environments at given spatial regions. These results suggest that spatial updating in naturalistic, nested environments does not occur for all environments at the same time. Implications for the hierarchical theory of spatial representations and the path integration theory of navigation are discussed. PMID- 12776751 TI - Priming and attentional control of lexical and sublexical pathways in naming: a reevaluation. AB - The authors report 3 naming experiments using J. D. Zevin and D. A. Balota's (2000) multiple prime manipulation. They used 2 sets of nonword primes (fast and slow) and low-frequency exception word primes to separate the effects of prime speed from those of prime type. The size of the regularity effect was unaffected by prime type. Relative to the low-frequency exception word prime condition, the frequency effect was reduced in the fast, but not in the slow, nonword prime condition. Lexicality effect size was reduced in both nonword prime conditions, a result consistent with the lexical checking strategy described by S. J. Lupker, P. Brown, and L. Colombo (1997). The authors suggest that these results are better explained in terms of S. J. Lupker et al.'s time-criterion account than J. D. Zevin and D. A. Balota's pathway control hypothesis. PMID- 12776752 TI - Articulatory duration in single-word speech production. AB - Three different speech production paradigms assessed C. T. Kello, D. C. Plaut, and B. MacWhinney's (2000) claim that the characteristics of speech production flexibly vary between staged and cascaded modes depending on task demand. All experiments measured response latencies and durations of single words without and with a response deadline. Experiment 1 used a picture-word interference task; Experiment 2 blocked pictures either by semantic category or by word-initial overlap; and Experiment 3 used a Stroop paradigm. In all cases, systematic effects of semantic and form relatedness were obtained on latencies but not on response durations. These results support the assumption that articulation, as assessed by response duration, is never influenced by central cognitive processes once a response has been initiated. PMID- 12776753 TI - Specific-word frequency is not all that counts in speech production: comments on Caramazza, Costa, et al. (2001) and new experimental data. AB - A. Caramazza, A. Costa, M. Miozzo, and Y. Bi (2001) reported a series of experiments demonstrating that the ease of producing a word depends only on the frequency of that specific word but not on the frequency of a homophone twin. A. Caramazza, A. Costa, et al. concluded that homophones have separate word form presentations and that the absence of frequency-inheritance effects for homophones undermines an important argument in support of 2-stage models of lexical access, which assume that syntactic (lemma) representations mediate between conceptual and phonological representations. The authors of this article evaluate the empirical basis of this conclusion, report 2 experiments demonstrating a frequency-inheritance effect, and discuss other recent evidence. It is concluded that homophones share a common word form and that the distinction between lemmas and word forms should be upheld. PMID- 12776754 TI - Effects of negation and situational presence on the accessibility of text information. AB - In 2 experiments, participants read narratives containing a color term that was mentioned either within the scope of an explicit negative or not, and with the described situation being such that the color was either present or not. Accessibility of the color term was measured by means of a probe-recognition task either 500 ms (Experiment 1) or 1,500 ms (Experiment 2) after participants read the sentence mentioning color. After the 500-ms delay, the accessibility of the color term was influenced by the structure of the sentence. After the 1,500-ms delay, the accessibility was influenced by the content of the described situation. These results are consistent with the view that comprehenders construct a linguistic representation of the text as well as a situation model in which only present properties are represented. An alternative account, according to which comprehenders only construct a perceptual simulation of the described situation, is discussed. PMID- 12776755 TI - How prior knowledge, WMC, and relevance of information affect eye fixations in expository text. AB - This study examined how prior knowledge and working memory capacity (WMC) influence the effect of a reading perspective on online text processing. In Experiment 1, 47 participants read and recalled 2 texts of different familiarity from a given perspective while their eye movements were recorded. The participants' WMC was assessed with the reading span test. The results suggest that if the reader has prior knowledge related to text contents and a high WMC, relevant text information can be encoded into memory without extra processing time. In Experiment 2, baseline processing times showed whether readers slow down their processing of relevant information or read faster through their relevant information. The results are discussed in the light of different working memory theories. PMID- 12776756 TI - Centripetal force draws the eyes, not memory of the target, toward the center. AB - Many observers believe that a target will continue on a curved trajectory after exiting a spiral tube. Similarly, when observers were asked to localize the final position of a target moving on a circular orbit, displacement of the judged position in the direction of forward motion ("representational momentum") and toward the center of the orbit was observed (cf. T. L. Hubbard, 1996). The present study shows that memory displacement of targets on a circular orbit is affected by eye movements. Forward displacement was larger with ocular pursuit of the target, whereas inward displacement was larger with motionless eyes. The results challenge an account attributing forward and inward displacement to mental analogues of momentum and centripetal force, respectively. PMID- 12776757 TI - Separating perceptual and decisional attention processes in the identification and categorization of integral-dimension stimuli. AB - Four observers performed matching, identification, and categorization with stimuli that varied along the integral dimensions: brightness and saturation. General recognition theory (F. G. Ashby & J. T. Townsend, 1986) was applied to quantify the separate influences of perceptual and decisional processes within and across tasks, with a focus on separating perceptual from decisional attention processes. Good accounts of the identification data were obtained from perceptual matching representation. This perceptual representation provided a good account of the categorization data, except when decisional selective attention to 1 stimulus dimension was required. Decisional selective attention reduced the attended-dimension perceptual variance relative to the unattended-dimension perceptual variance, with a larger reduction resulting when brightness, as opposed to saturation was attended. Implications for color vision research are discussed. PMID- 12776759 TI - Give yourself as a gift. PMID- 12776758 TI - Receiver operating characteristics in the lexical decision task: evidence for a simple signal-detection process simulated by the multiple read-out model. AB - This study provides further evidence for the hypothesis that performance in the lexical decision task is based on the output of a signal detection mechanism, as implemented by the multiple read-out model of word recognition (MROM; J. Grainger & A. M. Jacobs, 1996). We extend the MROM to allow predictions of receiver operating characteristics in a data-limited variant of the lexical decision task and show that the model provides accurate descriptions of the data. Our results challenge all models of word recognition that do not include a familiarity assessment mechanism. They also suggest that under data-limited conditions a deadline mechanism for generating "no" responses in the lexical decision task may not necessarily be functional. PMID- 12776760 TI - A call to arms. PMID- 12776761 TI - NSUCOM establishes bioterrorism preparedness center. PMID- 12776762 TI - Osteopathic manipulative treatment techniques preferred by contemporary osteopathic physicians. AB - Data presented in this study were gathered through a national mail survey of 3000 randomly selected osteopathic physicians. A total of 955 questionnaires were usable for analysis. Osteopathic physicians' likelihood of using eleven osteopathic manipulative treatment (OMT) techniques (articulatory, counterstrain, cranial, facilitated positional release, fascial ligamentous release, functional, high-velocity low-amplitude thrust, lymphatic, muscle energy, myofascial/integrated neuromuscular release, and soft tissue) was determined. The relative frequency of use from most (soft tissue) to least (cranial) used was also determined. Respondents were more likely to use direct techniques than indirect or direct-indirect techniques. Demographic variables of gender, age, and specialty training were found to be related to the techniques used most. Female osteopathic physicians and older osteopathic physicians were more likely to use indirect techniques, whereas male and younger physicians preferred direct techniques. Moreover, OMT specialists used a broader range of techniques than other osteopathic physicians, and family physicians were more apt to use high velocity low-amplitude thrust than other primary care or non-primary care osteopathic physicians. These results not only have implications for curricular planning in all phases of osteopathic undergraduate medical education, graduate medical education, and continuing medical education programs, but also for research on the quality and effectiveness of various OMT techniques. PMID- 12776763 TI - Using National Medical Care Survey data to validate examination content on a performance-based clinical skills assessment for osteopathic physicians. AB - Patient characteristics, chief complaints, and diagnoses can be used to specify the examination content for performance-based assessments of clinical skills. The purpose of this investigation was to explore osteopathic and allopathic medical practice patterns and to provide summary statistics that can be used to delimit potential assessment content areas for a clinical skills assessment targeted at osteopathic physicians. Analyses of the National Ambulatory Medical Care Survey indicated that the types of patients seen by osteopathic and allopathic physicians in office-based settings are somewhat different. Furthermore, the reasons that patients seek care, and accompanying diagnostic outcomes, can vary by physician type. These differences suggest that from a content perspective, a performance-based clinical skills evaluation targeted at osteopathic physicians should be characteristically different from one designed for allopathic physicians. PMID- 12776764 TI - A myoelectric model for thoracic spinal motion dynamics during clinical rotation tests: Part 2. Bilateral segmental motor behaviors. AB - In part 2 of their report, the authors continue to evaluate myoelectric data obtained from spinal motion dynamics involved in clinical rotation tests. They add to the ipsilateral regional analysis of motor performance as previously presented and analyze the total bilateral myoelectric activity gathered concurrently at individual thoracic vertebral segments during simultaneous rotation left and right motion tests. The authors' hypothetical consideration concerns the nature of composite behaviors at these vertebral segments during active and passive motions and the role that postural dynamics play in movement function. They consider these concepts in the context of the study's experimental design and also within the broader concept of the osteopathic musculoskeletal examination. Results revealed pronounced similarity in individual spinal electromyographic patterns whether motions were volitional or physician induced. Analysis demonstrates the bell-shaped myoelectric behavior pattern originally reported in part 1. The authors also discuss a functional model for this myoelectric activity involving a helical spinal motor pattern with a focal area of transition that is dynamic in response to postural and motion demands. PMID- 12776765 TI - Evaluating the impact of stress on systemic disease: the MOST protocol in primary care. AB - Mental stress has an enormous impact on physical health. This impact commonly manifests as headache, muscle tension, acne, peptic ulcer disease, or a compromised immune system. Stress is also associated with more serious adverse effects, such as cardiovascular disease and exacerbations of rheumatoid arthritis and systemic lupus erythematosus. As these effects are far-reaching, it is important for primary care physicians to identify and manage the symptoms of mental stress in their patients. This is increasingly possible with office-based mental stress testing, which uses cardiovascular markers to identify patients who are overresponders to mental stress, and, thus, at risk for stress-induced disorders. Mental stress in this population can be managed with nonpharmacologic and pharmacologic interventions to improve patients' responses to stress and decrease morbidity and mortality associated with this condition. PMID- 12776766 TI - Dilated cardiomyopathy after electrical injury: report of two cases. AB - The specific etiologic factor and pathogenesis of most dilated cardiomyopathies have yet to be described definitively. Hypotheses of the etiologic factor of idiopathic dilated cardiomyopathy (DCM) abound. This report describes two patients with electrical injury in whom DCM developed after the electrical insult in the absence of other precipitating causes. Further histologic examination of myocardial tissue after electrical injury may reveal clues regarding the pathophysiology behind electrically induced DCM. Because electrical injury may be associated with myocardial dysfunction, short- and long-term evaluation of left ventricular function may be warranted. PMID- 12776767 TI - Payment for laboratory services. PMID- 12776769 TI - Lessons learned in student recruiting. AB - Nationally, clinical laboratory science programs are struggling for student applicants. Major challenges facing the laboratory profession include: 1) low salaries, 2) lack of public awareness, and 3) the myriad of career choices for new graduates. Increasing public awareness and actively recruiting students can overcome one of these challenges. This paper focuses on the successful student recruiting lessons learned at the University of Utah Medical Laboratory Science Program. Specific indicators show increased interest and activity for this program of study. PMID- 12776768 TI - Dyslipidemia prevalence in a laboratory initiated screening program. AB - OBJECTIVE: Evaluate utilization and diagnosis rates in a self-pay, self-referred screening program for dyslipidemia. DESIGN: 301 patients self-referred to the clinical laboratory for lipid testing in a two-year period. The patient population that participated was characterized in terms of insurance status, gender, age, and known cardiovascular risk factors. Lipid profiles were characterized as measured by total cholesterol, triglycerides (TGs), low-density lipoprotein cholesterol (LDL), high-density lipoprotein cholesterol (HDL), and total cholesterol to HDL risk factor. SETTING: Clinical laboratory in an academic medical center. PATIENTS: Data from all patients that self-selected for screening were included. INTERVENTIONS: Immediate laboratory results with both verbal and written interpretations and recommendations were provided to the patients. MAIN OUTCOME MEASURES: Age, gender, insurance status, number of known risk factors, and lipid profiles in the subject group. RESULTS: The mean age of participants was 57 years. Men (197) outnumbered women (104) by almost 2:1; most (94%) had health insurance. At presentation, 44% of the patients had more than one risk factor for coronary heart disease (CHD). 151 individuals (50%) had lipid findings that would require at least dietary intervention by NCEP guidelines. CONCLUSION: A self-pay, self-referred screening program for lipid disorders is an effective means of improving screening and diagnosis rates. Patients with insurance were willing to pay for the convenience offered and men in particular were more likely to self-refer than women, independent of previous knowledge of risk factors or lipid disorders. PMID- 12776770 TI - A comparison of INRs after local calibration of thromboplastin international sensitivity indexes. AB - There are approximately 300 reagent/instrument combinations for performing prothrombin times/international normalized ratios (PT/INR) in the United States. Manufacturers and laboratories continually struggle to ensure that the International Sensitivity Index (ISI) of their thromboplastin is accurate for assaying PT/INR. OBJECTIVE: This study reports the feasibility of a new method to locally calibrate ISI of thromboplastin using the mechanical STA automated coagulation analyzer (Diagnostica-Stago Inc.) and two photo-optic coagulation analyzers, the BCS (Dade-Behring) and CA-540 (Sysmex). DESIGN: Neoplastine CI+ (CI+) (Diagnostica-Stago Inc); Thromboplastin C+ (TC+); Thromborel S (TRS); and Innovin (I) (Dade-Behring) were used in this study. A mean normal PT (MNPT) was determined for each reagent/instrument combination using samples from 25 normal individuals. Manufacturer instrument specific ISI values were not available for the STA with TC+, TRS and I. The CA540 had no ISI value for CI+ and the BCS system had no manufacturer assigned ISI values for TC+ and I; generic photo-optic and mechanical ISI manufacturer values were used for these two systems. Local on site calibration was performed using frozen plasma calibrators to determine ISI values for each thromboplastin. Post-calibration, 95 patient samples were assayed for each reagent/instrument system combination using the manufacturer ISI and the local calibrated ISI to determine the INR result. PATIENTS: Patients from whom samples were obtained included five with a lupus anticoagulant, 30 on heparin therapy, and 60 on coumadin therapy. RESULTS: Differences between manufacturer versus local calibrated ISI ranged from 0.9% to 18.9% for normal sample INRs and from 0.8% to 16.4% for patient sample INRs. The number (or proportion) of patient specimens with clinically significantly different INR values (>10.0% difference) ranged from zero for several reagent combinations to more than half (or >50.0%) of those tested for several other combinations. CONCLUSION: Our results indicated that by locally calibrating ISI values, each laboratory may eliminate variability and guesswork between different reagent/instrument systems for ISI values when performing PT/INR assays and potentially improve the clinical accuracy of their patients' PT/INR results. PMID- 12776771 TI - Fetal fibronectin. AB - OBJECTIVES: The objectives of this review paper are to: describe the fetal fibronectin assay, its purpose, and clinical significance; evaluate the sensitivity and specificity of the fetal fibronectin test; describe the specimen collection and measurement of the fetal fibronectin test; and present the advantages and disadvantages of incorporating fetal fibronectin testing in routine prenatal care. DATA SOURCES: Current literature. DATA SYNTHESIS: Fibronectin proteins function in plasma and extracellular matrix in cell adhesion and migration. Recently, a fibronectin protein has been evaluated and proposed as a predictor of preterm delivery. A simple, qualitative assay detects this protein, fetal fibronectin, in cervicovaginal secretions of women who are at risk for or have symptoms of preterm delivery. The test is positive when there has been a rupture in the membranes attaching the fetus to the uterus, thus indicating pending preterm delivery. Sensitivity and specificity studies have been performed to evaluate its reliable prediction of preterm delivery. CONCLUSION: Evaluation of sensitivity and specificity studies document that the fetal fibronectin test predicts preterm delivery. For symptomatic women, a sensitivity of 89% and specificity of 86% was found. PMID- 12776772 TI - Perceived barriers to articulation: institutional characteristics. AB - It has been generally acknowledged that a number of obstacles, or barriers, exist in the articulation process. Based on literature review, student characteristics as well as institutional characteristics may act as barriers. This paper focuses on institutional characteristics. The changed mission of the community college and a lack of standardization of curricula between two-year and four-year institutions of higher education have been identified as barriers to articulation. Suggested reforms are described. PMID- 12776773 TI - Knowledge fields and inner patterns in clinical laboratory science. AB - OBJECTIVE: The purpose of the study was to clarify the knowledge base of clinical laboratory science (CLS). This research was motivated by questions concerning the knowledge base itself and its abilities to meet the demands of reality. The following questions were therefore asked to achieve the purposes of the study: What are the knowledge fields and inner patterns in CLS? Which research objects could CLS focus on in order to promote development in practice, education, and research? DESIGN: The findings of the study were arrived at by means of hypothetical-deductive approach and inductive, content analytical strategy. The journal Clinical Laboratory Science of the American Society for Clinical Laboratory Science (ASCLS) provides the source material for the analysis. SETTING: Abo Akademi University, Faculty of Social and Caring Sciences. RESULTS: The findings of the study are discussed in the light of starting points of the theory of science and lead to nine hypotheses concerning CLS. CONCLUSION: The purpose of the present study was to create clarity in CLS as a science of its own. This has been achieved by capturing and describing facts and qualities, and thereafter presenting fundamental hypotheses in CLS. The results of this study give a thought structure for continued development and deepening within the theory and practice of CLS. PMID- 12776774 TI - A procedure for the detection of Stealth adulterant in urine samples. AB - Stealth is an adulterant that is advertised as not only preventing a positive drug test in urine, but also to be undetectable by currently available adulteration testing. It has previously been described as a peroxidase and peroxide that is added to urine for the sole purpose of preventing a positive drug test. The product was found to have a significant impact on the ability to detect several drugs of abuse, however, detecting the presence of the adulterant in urine had not yet been reported. A simple procedure to detect the presence of this adulterant in urine was developed. This simple color test procedure using commercially available reagents commonly used in clinical laboratories is based on the use of a chromogen to detect the peroxidase reaction in urine samples. If Stealth is present in the urine, the test sample will show an immediate color change from clear to dark brown. This qualitative test can also be adapted for use with a spectrophotometer or autoanalyzer. PMID- 12776775 TI - Developments in component therapy: novel components and new uses for familiar preparations. AB - Over the years, the significant role of blood components in treating certain diseases or conditions has been recognized. The use of these components has expanded as patients undergo chemotherapy for bone marrow ablation and require short-term component support. On the other hand, these transfusions can cause reactions ranging from mild to severe. Despite advances in serological testing for infectious disease agents, the risk of infectious complications from transfusion still remains. In addition, newly identified agents that may be transmitted via transfusion are constantly identified. The cellular components most people are familiar with include packed red blood cells (PRBC), washed PRBC, leukoreduced PRBC, and pooled or apheresis platelets. Plasma products such as fresh frozen plasma (FFP) or crytoprecipitated anti-hemophiliac factor (CRYO), on the other hand, may not be as familiar. As our understanding of how the immune system functions and as technology has progressed, specialized components or manufactured products such as blood substitutes have been advanced as remedies to some of the complications with component transfusion or to meet the ever increasing need for these products. In this article we will focus on some of the new uses of common components and uncommonly used or newly developing components. We will discuss their origins, composition, and the conditions or diseases they are used to treat. These components include: donor leukocyte infusions, dendritic cell vaccines, blood substitutes, novel platelet products and substitutes, intravenous immunoglobulin (IVIG), fresh frozen plasma and cryosupernatant in therapeutic plasma exchange. The variety of products and conditions reflect the ever-expanding role of immunohematology in the treatment of disease. PMID- 12776776 TI - The hard work of advocacy. PMID- 12776777 TI - Hereditary deficiencies of antithrombin III, protein S, and the protein C pathway in Jordanian thrombosis patients. AB - Hereditary thrombophilia is caused by various inherited disorders. Most lead to a familial tendency to recurrent venous, not arterial, thrombosis, usually at a young age, and with spontaneous onset. Most of the genetic defects known today affect the function of natural anticoagulant pathways, in particular, the protein C system. In this study, 602 (265 female, 337 male) patients with suspected thrombosis, arterial or venous, were referred to King Hussein Medical Center in Amman, Jordan. The prevalence of hereditary deficiencies of antithrombin (AT), protein S (PS), and protein C (PC) were studied over a seven-year period (1993 2000). Activated protein C (APC-R) resistance subjects were studied over four years (1996-2000). The mean age was 30 years in females and 42 years in males. A diagnosis was established in 22.4% (n = 135) of the subjects (20.3% venous, 2.1% arterial). Protein C deficiency was found in 3.8%, protein S deficiency in 2.3% and antithrombin deficiency in 1.4% of our sample group. An APC-R problem was seen in 23.0% (n = 89) of the surveyed population. Out of the APC-R patients, 75.0% had the DNA analysis of a factor V Leiden mutation present. Of the subjects found to have the mutation 87.0% were heterozygous and 13.0% were homozygous. These results confirm that APC-R, as a result of factor V Leiden mutation, is the most prevalent cause of thrombosis, and thrombophilia is related to venous, not arterial, thrombosis. PMID- 12776778 TI - Cultural competency in the laboratory. AB - Cultural competency incorporates an awareness of different cultures, and sensitivity to the differences of cultures which results in appropriate behaviors with other cultures. White non-Hispanics currently comprise almost 73.6% of the total United States population, but this percentage will decrease to less than 53% by 2050. Laboratory practitioners must become culturally competent to deal effectively with the changing demographics. Cultural diversity will impact the laboratory workforce and the patients it serves. Laboratory professionals must become skilled in communication with all patient populations served, be aware of personal biases, and be willing to do whatever is necessary to achieve cultural competency. Skills must be integrated into education and training processes to prepare the laboratory professional to work effectively in and with different cultures. PMID- 12776779 TI - ABO discrepancy in a multiple myeloma patient: a case study. PMID- 12776780 TI - Clinical utility of the IRF: assessment of erythroid regeneration following parvo B19 infection. AB - Parvo B19 (Fifth disease) is an erythrotropic virus which attaches through the 'P' globoside receptor on the surface of human red blood cells and precursors. This typically benign viral infection can cause a transient aplastic anemia in patients with underlying red cell disorders. In this case, a two-year-old child presents with severe aplastic anemia without evidence of underlying disease. Erythroid regeneration is monitored through the use of the immature reticulocyte fraction (IRF) and is demonstrated by the presence of high and medium fluorescence reticulocytes in the peripheral blood three to five days prior to the peak in absolute reticulocytes. PMID- 12776781 TI - HIV-1 antiretroviral resistance testing laboratories. AB - OBJECTIVE: To identify and to describe the genotyping and the phenotyping testing practices of U.S. laboratories performing patient HIV-1 antiretroviral resistance testing. DESIGN: A self-report 44-item mailed questionnaire. PARTICIPANTS: Laboratories potentially performing HIV-1 antiretroviral resistance testing. MAIN OUTCOME MEASURE: Descriptive study. RESULTS: Of 236 laboratories surveyed, 165 (69.9%) returned completed surveys, but only 23 performed HIV-1 antiretroviral resistance testing. Most were university hospitals (47.8%) or independent laboratories (26.1%). All 23 laboratories used genotypic methods, while nine (39.1%) used both genotyping and phenotyping. Most testing was used for clinical trials or laboratory research. The amount of patient information collected by laboratories varied, as did their type of quality assurance measures. Variation was found with regard to: testing volume, testing experience, testing reasons, testing methods availability, testing controls, specimen treatment, and storage stability. CONCLUSIONS: Due to variation in practices in this area of patient testing, it may be advantageous for laboratory professionals to reach a consensus on what is the most acceptable. PMID- 12776782 TI - CLS competencies expected at entry-level and beyond. AB - OBJECTIVE: The study was undertaken to assess educators', practitioners', and managers' perceptions of the future job expectations of clinical laboratory scientists (CLSs) and their opinions on the skills that are expected of CLSs at entry-level and with experience. DESIGN: Survey participants were given a list of 44 competencies related to clinical laboratory science (CLS) practice and were asked whether they would expect a graduate of a respected CLS program to perform each competency in one of three educational categories: the first year of practice, with three to five years of experience but no additional education, or with three to five years of experience plus additional education. The competencies were subclassified into one of four major management functions: laboratory operations, human resource management, financial operations, or communications/consultation. Surveys also included eight Lickert-type questions designed to assess the respondents' opinions on the future job expectations of CLS practitioners. PARTICIPANTS: The sample for the survey included 280 directors of CLS educational programs, 600 managers randomly selected from the Clinical Laboratory Management Association (CLMA) membership, and 600 practitioners randomly selected from the American Society for Clinical Laboratory Science (ASCLS) membership. MAIN OUTCOME MEASURES: The percent of respondents selecting each educational category was tabulated and each competency was assigned to one educational category based onthe highest percent of respondents selecting that category. The means of the responses to the Lickert-type questions were calculated for all respondents and for each group of respondents (educators, managers, and practitioners). RESULTS: Response rates of 58% (educators), 28% (practitioners), and 39% (managers) were obtained. Of the 44 competencies in the survey, four were expected at career-entry, 17 were expected of CLS graduates with work experience but no additional education, and 23 were expected of CLS graduates with experience plus additional education. Competencies expected in the first year of practice were primarily scientific and technical. With three to five years of practice and no additional education, the expectations for practitioners were primarily in laboratory operations and communications/consultation areas. The majority of the human resource management and financial operations competencies were expected with three to five years of practice and additional education. All participants agreed that CLS staff-level practitioners need more management and administrative skills and that, in the future, CLS practitioners will spend less time performing laboratory tests and more time solving problems. CLS managers were more positive than CLS educators in response to statements asserting that CLT practitioners and non-certified personnel will have an increased role in the laboratory in the future. CONCLUSION: This study suggests that extensive laboratory operations and communication skills are expected of CLS graduates without any additional education beyond their CLS programs. CLS educators should adequately address those areas in the curriculum. Competence in other non-technical skills may not be expected without the benefit of post-baccalaureate education and in these areas, CLS programs can provide a foundation for future learning. PMID- 12776783 TI - Newborn screening: an overview. AB - The ethical considerations and the criteria for inclusion of a test to a newborn screening program have remained constant since testing began in the 1960s. Does the test identify a treatable disorder with significant incidence to pose a public health risk and warrant testing all babies in that state or territory? Technological advances in testing, particularly with the improvement of tandem mass spectrometry techniques and the advent of DNA testing for the specific gene mutations, have expanded our understanding of many inherited metabolic diseases. These mostly autosomal recessive disorders went under-diagnosed by the medical community for many years. This was partly due to the notion that the incidence of inherited metabolic diseases was quite rare and that many so-called birth defects, or unexplained infant deaths, were not associated with any known metabolic disorders. Public health departments, as part of their newborn health programs, offer some newborn screening to all infants born within their jurisdiction. Two tests, those for phenylketonuria (PKU) and congenital hypothyroidism are universally mandated (51/51 juristictions). The next highest frequency tests are for galactosemia and sickle cell disease (50/51), with up to thirty tests available in some states. However, the authority as to which tests are included resides with the local state government, either as a matter of law or as a matter for the public health department. As these matters become more complex, many public health officials and pediatric healthcare practitioners urge the Federal government to become involved and develop national guidelines in an effort to streamline the process and decrease the existing inconsistencies between states. For many laboratorians, the collection of newborn screening blood spot samples is the extent of their involvement in newborn screening programs. The many facets of these programs, the status of newborn screening in the United States, and the incidence and description of selected inherited disorders are explored. PMID- 12776784 TI - Newborn screening: new developments in a proven field. PMID- 12776785 TI - Lesions induced in broiler chickens by cyclophosphamide treatment. AB - Cyclophosphamide (CY) is an immunosuppressant and inhibits cell division by alkylating nucleic acids. This study was performed to determine the toxic effects of CY treatment in broiler chickens. One-d-old White Plymouth Rock broiler chickens were treated with either 4 mg/d CY or phosphate buffered saline (PBS) for 4 consecutive days. Mortality of the CY-treated chickens was 65.4% (34/52), while that of the PBS-treated chickens was 10% (5/50). CY-treated chickens were significantly smaller and had delayed feathering and lower relative bursal weight compared to those PBS treated. Histologically, the lymphocytes were markedly depleted in the spleen and bursa. Bone marrow was severely hypocellular and replaced by adipose tissue. Diffuse vacuolation of hepatocytes with individual cell necrosis was present in the liver. There was segmental necrosis of lining epithelial cells of renal tubules with eosinophilic material and necrotic detritus within the lumens. The lesions in the liver, kidney and bone marrow were transient and repaired by 2 w of age. PMID- 12776786 TI - The comparative efficacy of yohimbine and atipamezole to treat amitraz intoxication in dogs. AB - This study compared the efficacy of yohimbine with atipamezole, a new alpha2 adrenergic antagonist, to treat canine amitraz intoxication. Thirty dogs were divided equally into 3 groups (A, AY, and AA). Group A received 2.5% amitraz iv at 1 mg/kg; Group AY received the same dose of amitraz followed 30 min later by 0.1 mg/kg (2 mg/mL) yohimbine iv; and Group AA received the same dose of amitraz followed 30 min later by 0.2 mg/kg (5 mg/ mL) atipamezole iv. Temperature, heart rate, respiratory frequency, mean arterial pressure, degree of sedation, mean time of tranquilization and diameter of pupils were monitored for 360 min. Sedation, loss of reflexes, hypothermia, bradycardia, hypotension, bradypnea and mydriasis were observed in Group A, with 3rd eyelid prolapse, increased diuresis and vomiting in some animals. Yohimbine reversed all alterations induced by amitraz, but induced significant cardiorespiratory effects such as tachycardia and tachypnea. Atipamezole was a useful antagonist for amitraz, with less cardiorespiratory effects, suggesting its potential role as an alternative treatment of amitraz intoxication in dogs. PMID- 12776787 TI - Cadmium and lead accumulation in cattle in NW Spain. AB - This study investigated if differences in nutritional management (linked to feed composition and feed intake) could determine differences in the toxic metal accumulations in beef and dairy cattle in NW Spain. Cadmium and lead concentrations were determined in the liver, kidney and muscle of 57 cows (3-16 y o) by inductively coupled plasma optical emission spectroscopy. Dairy cattle accumulated significantly higher cadmium and lead residues in liver (51.6 and 43.7 microg/kg fresh weight) than did beef cattle (35.7 and 14.7 microg/kg). In the kidney and muscle, cadmium and lead residues were similar in both dairy and beef cattle. The higher metal accumulations in the liver of dairy cattle could be related not only to higher dietary intake, but also to the higher hepatic metabolism associated with milk production. PMID- 12776788 TI - Placental and fetal toxicity of albendazole sulphoxide in Wistar rats. AB - This work characterized the effects of albendazole sulphoxide (ABZSO) on placental and fetal parameters in Wistar rats on day 20 of gestation. ABZSO was fed in laboratory chow at 0, 2.5, 5, 10, 20 or 30 mg/kg/d from day 6 to 15 of gestation to pregnant rats. Data of resorptions, placental and fetal characteristics and fetal skeletal malformations were recorded. Resorption percentages in the 20 and 30 mg/kg/d groups were significantly higher compared to the control group. Placentas of ABZSO-treated rats had lower weight and smaller size than untreated rats. The fetal weight and size were lower in the 5 mg/kg/d dose compared to no treatment. In the 5, 10 and 20 mg/kg/d groups, reductions in ossification process were observed. ABZSO induced malformations and/or fetal death when orally administered to pregnant rats. This data contributes to characterization of the reproductive toxicity of ABZSO, the main active metabolite of albendazole. PMID- 12776789 TI - Catastrophic cattle loss to low larkspur (Delphinium nuttallianum) in Idaho. AB - Low larkspur (Delphinium nuttallianum) is a toxic plant found on many western US rangelands. Episodes of fatal poisoning are often related to the abundance and toxicity of the low larkspurs. This report documents the fatal intoxication of 53 cattle in a herd of 404 animals in south-central Idaho during Spring 2002. Low larkspur had a toxic alkaloid concentration of 8.26 mg/g. Several factors were involved in a loss of this magnitude, including climatic conditions ideal for low larkspur growth, poor grass growth during the spring, the high alkaloid concentration in larkspur, and the deaths of many cattle while being moved from the pasture. It is important for livestock owners and managers to recognize toxic plants growing in their area. If there is an unusual abundance of larkspur or other toxic plants, correct identification can provoke caution in grazing management. PMID- 12776790 TI - Hyperthermic syndrome in dairy cattle associated with consumption of ergots of Claviceps purpurea in southern Brazil. AB - A hyperthermic syndrome (HS) associated with consumption of ration contaminated with Claviceps purpurea is described in 4 dairy cattle herds from different counties in the state of Rio Grande do Sul, southern Brazil. Outbreaks of this mycotoxicosis occurred during the summer of 1999-2000 and autumn of 2002 when 25% to 70% of cattle from those herds were affected. The disease occurred after the introduction of a new batch of feed contaminated with sclerotia of C. purpurea. First clinical signs were observed up to 3 mo after the ration was introduced and consisted of pyrexia, intense salivation and drooling, open-mouth, difficult breathing, in some cases with the tongue protruded, decrease in milk yield, decrease in food intake and weight loss. Affected cattle compulsively sought shade or water ponds. Signs were more pronounced during daytime. The severity of the clinical picture varied according to changes in environmental temperature during the day. There was complete recovery approximately 60 d after feeding with the contaminated ration was discontinued. Abortions occurred during the outbreak and soon after the remission of the clinical signs associated with HS. In one of the herds, reproductive performance and fertility were seriously impaired even after clinical signs of HS completely disappeared. Diagnosis was based on epidemiology, clinical signs and pathology. The pathogenetic mechanisms involved in the occurrence of HS are briefly discussed. PMID- 12776791 TI - Intoxication by cholinesterase inhibitors versus opioid intoxication. AB - A 47 y-old male shopkeeper from a rural area ingested an unknown substance while under the effects of ethylic alcohol. He was admitted at the University Hospital of the Andes in generally poor condition with a cholinergic syndrome. An erroneous diagnosis of acute pulmonary edema and opioid intoxication was reached. The value of a patient's history (background) and careful evaluation of the physical examination findings without underestimating critical clinical signs are very important when handling a clinical intoxication. PMID- 12776792 TI - Spontaneous poisoning by Dodonea viscosa (Sapindaceae) in cattle. AB - This communication reports a southern Brazilian outbreak of acute hepatic insufficiency in which 14 dairy animals died after consumption of Dodonea viscosa. The clinical course lasted a few hours and was characterized by apathy and staggering disorders. The main postmortem findings occurred in livers as enhanced lobular patterns and hepatocellular necrosis that ranged from centrilobular to massive. Similar clinical and pathological effects were experimentally induced by dosing a heifer with 30 g/kg of the plant's green leaves. PMID- 12776793 TI - Alpha-methyltryptamine revisited via easy Internet access. AB - Alpha-methyltryptamine (AMT) is a synthetic hallucinogenic indolealkylamine, that was initially studied as a monoamine oxidase inhibitor and used as an antidepressant. Although a popular hallucinogen in the 1960's, use of this drug is currently uncommon. We report a patient with sympathomimetic features who was found to have ingested AMT. PMID- 12776794 TI - The difficulty of risk assessment for hepatic injury associated with supra therapeutic acetaminophen use. AB - We examined historical and laboratory features to identify patients who do not experience hepatic injury when presenting with a history of repeated supra therapeutic acetaminophen use by performing a retrospective, double cohort comparison over almost 5 y. One cohort included patients suffering severe hepatic injury after supra-therapeutic acetaminophen ingestion; the control cohort included all patients in a geographically limited population with repeated supra therapeutic acetaminophen ingestion who did not experience severe hepatic injury. Demographics, baseline health, medications, ethanol consumption, acetaminophen dosage, reason for excessive acetaminophen dosing, acetaminophen concentration at presentation, presentation and peak hepatic enzymes, N-acetylcysteine (NAC) treatment, highest measured AST and outcome were abstracted from 1114 possible patient contacts. Twenty two of these met all inclusion criteria. Ten suffered severe hepatic injury, 12 did not. The actual acetaminophen dose consumed over a period of time was difficult to establish, but the historic quantity ingested exceeded 20 g in the majority of patients in both groups. High ethanol use was common in both groups. All patients who ultimately suffered severe liver injury presented with some injury. No patient who presented without some injury suffered severe injury. None of these patients were treated with NAC. Two patients who presented with minor injury (1 of whom received NAC), did not progress to severe injury. Future investigations of the cause of liver injury in patients exposed to supra-therapeutic doses of acetaminophen will require larger patient numbers. The lack of injury in similarly exposed patients underscores the peril of attempting to assess the role of risk factors without an appropriate control group. Our experience suggests history is unlikely to be clinically useful in predicting risk of injury. Future risk assessment studies should focus on objective presentation features like presence or absence of injury or serum acetaminophen levels. PMID- 12776795 TI - Acute zonisamide overdose: a death revisited. AB - Overdose experience with the anti-epileptic agent zonisamide (Zonegran, ZNS) is limited. We present a case of suspected zonisamide overdose previously reported in abstract form, manifesting as seizures, dysrhythmias and death of an 18-y-o woman after single ingestion of 4.8 g ZNS in a suicide attempt. The patient experienced multiple generalized tonic-clonic seizures and copious airway secretions. En route to the emergency department, she sustained cardiopulmonary arrest from which she was resuscitated. CT scan of the head approximately 24 h after initial presentation revealed massive cerebral edema with tonsillar herniation; brain death was confirmed. Serum ZNS was 44 mg/L, and the case was officially deemed death secondary to ZNS overdose. Subsequently, a comprehensive urine drug screen demonstrated mirtazepine, diphenhydramine metabolites, and caffeine. Due to confounding factors, a definitive causal role for ZNS in this overdose death cannot be made, despite the county coroner's ruling. PMID- 12776796 TI - Effects of a saccharin and cyclamate mixture on rat embryos. AB - Sodium saccharin (NaS) and calcium cyclamate (CaC) are artificial sweeteners widely used in food and drink. To evaluate their toxicological effects on preimplantation mammalian embryos, pregnant rats were gavaged with 1.65 mg NaS/kg bw + 3.85 mg CaC/kg bw (DI) or 6.6 mg NaS/kg bw + 15.4 mg CaC/kg bw (D2) on days 1, 2, 3 and 4 of pregnancy (positive vaginal smear = day 1). The female rats were killed on day 5 of the pregnancy (GD 5), maternal organs weighed, and the blastocysts collected, counted and evaluated for gross morphology, cell number and mitotic index. There was no alteration in maternal organ weights, but there was an increase of the cell number/embryo in the dams treated with that NaS + CaC mixtures (D1 = 37.20 +/- 7.96; D2 = 37.26 +/- 10.90) compared to control group (32.24 +/- 6.73). Embryos whose dams were exposed to NaS + CaC may have adapted for implantation into the uterus but more studies are needed to demonstrate this mechanism of action. PMID- 12776797 TI - Poisoning of livestock in oregon in the 1940s to 1960s attributed to corynetoxins produced by Rathayibacter in nematode galls in chewings fescue (Festuca nigrescens). AB - Tunicaminyluracil antibiotics, similar to the corynetoxins produced by Rathayibacter toxicus in Australia and South Africa, were found in old nematode seed-galls from Festuca nigrescens from New Jersey (USA) and New Zealand (NZ). The toxin profiles from the NZ and USA galls were similar to each other, but differed from those produced by R toxicus from Australia and South Africa, suggesting that a geographical variant of R toxicus or closely related species may be involved. The NZ galls gave a positive response to a R toxicus-specific monoclonal antibody assay, albeit a considerably weaker response than that seen with Australian R toxicus galls, but the older USA galls were negative, possibly due to deterioration of the antigen. From these findings, it is postulated that livestock deaths associated with the feeding of nematode and bacterial infected screenings of F nigrescens in Oregon, USA, in the 1940s to 1960s were caused by corynetoxin-like toxins produced by the bacterium. PMID- 12776798 TI - Foamy macrophages in the liver of cattle fed Brachiaria brizantha hay. AB - Liver and lymph nodes injuries characterized by clusters of foamy macrophages, some of them containing birefringent crystals, were observed in cattle fed on Brachiaria brizantha hay. The cattle were from an experimental group poisoned with Senecio brasiliensis known to cause hepatic fibrosis and hepatocyte megalocytosis. One of the animals developed photosensitivity but the exact cause wasn't determined since both plants were fed. The foamy macrophages were present from the 30th d of feeding. Early appearance of these lesions may be particular to the animal specie used or due to the presence of both toxic plants. PMID- 12776799 TI - Tomorrow today: plant-made pharmaceuticals. PMID- 12776800 TI - Pralidoxime hydrolysis of thiocholine esters. PMID- 12776801 TI - Pharmacological approaches to the management of alcohol addiction. AB - Alcohol abuse and alcoholism represent a world-wide problem, both from a medical and a social point of view. In the past the therapy for patients affected by alcoholism was based mainly on the psychological approach. In recent years the use of pharmacotherapy together with psychosocial interventions have enhanced the percentage of success in maintaining alcoholic patients in remission. The present review discusses the main drugs experimented both in preclinical and clinical studies. Pharmacotherapy of alcohol dependence seems to be effective in both alcohol-related emergencies and prevention relapse. However, pharmacotherapy should not be considered as the only form of treatment but as an integrated part of a multimodal approach including psychological and social support. PMID- 12776802 TI - New born chicks can serve as an experimental animal model for human campylobacteriosis. AB - Campylobacter enteritis is an emerging food borne zoonotic disease. Improperly cooked chicken serve as a potential source for this infection. Diarrheogenic potential of Campylobacter jejuni is tested either by in-vivo rat ileal loop (RIL) test or by molecular methods. This study reveals that 3-day-old chicks can serve as an animal model for toxigenic C. jejuni. PMID- 12776803 TI - The abortive form of Bourneville-Pringle syndrome. AB - BACKGROUND/METHODS: To present a 26-year-old woman affected by the abortive form of Bourneville-Pringle syndrome. To our knowledge, this disease is unusual since only very few cases have been reported in the scientific literature at this time. RESULTS/CONCLUSIONS: Visual acuity was 20/20 in both eyes. No relevant ocular abnormalities were observed excepting two retinal hamartomas, a smaller one in the nasal midperiphery of the right eye and a larger one located along the super temporal retinal vessels of the left eye. Classical signs of Bourneville-Pringle disease, such as mental retardation and epilepsy, were absent whereas a slight facial adenoma sebaceum and renal cysts represented the solely systemic manifestations of the disease. This case report confirms that retinal phakomata are a typical manifestation of Tuberous Sclerosis, even in the absence of a detected involvement of the brain. PMID- 12776804 TI - Warthin's tumor of parotid gland: treatment of a retroneural lesion by enucleation. AB - The papilliferous cystadenomas lymphomatosum or Warthin's tumor is a relatively rare and generally benign neoformation whose incidence is second only to the pleomorfic adenoma. 84% of all Warthin's tumor strikes the parotid gland. We report the two cases that come to our observation paying particular attention to the second one in which occurred a synchronous bilateral manifestation, a rarity in the clinical circle. We want to highlight that enucleation should be the preferred treatment in most cases due to extremely small dimension of Warthin's tumor. Moreover it is possible to perform this kind of surgical treatment even in the case of a retroneural lesion without causing any damage to the facial nerve and its main branches. As regards complications and sequelae we noticed neither a permanent nor a temporary injury of the facial mimicry and sensibility. PMID- 12776805 TI - Carvedilol: something else than a simple betablocker? AB - Carvedilol is a cardiovascular drug of multifaceted therapeutic potential, with beta-blocker and vasodilatative activity. These actions confer to the above mentioned betablocker some beneficial properties on several processes involving cardiovascular system. Carvedilol provides haemodynamic, antiischemic, antiproliferative and antiarrhytmic benefits, for its antioxidant neurohumoral and electrophysiological effects. All these actions provide the basis for usefulness of the drug in the treatment of hypertension, coronary heart disease, and congestive heart failure. In this review we report the beneficial properties of Carvedilol and we analyze the rational clinical use of this betablocker taking special attention on recent clinical trial in heart failure where it appears an evidence supporting an important, favourable effect of the drug. PMID- 12776806 TI - Cyclosporine bioavailability of two physically different oral formulations. AB - To assess the comparative bioavailability of two cyclosporine capsule products with different pharmaceutical formulation an open randomized two-period cross over study was conducted in 24 healthy volunteers. Our results, obtained from cyclosporine HPLC determination onto the whole blood samples collected, show that the test cyclosporine non-SMEDDS formulation was not bioequivalent cyclosporine SMEDDS formulation due to a statistically significantly lower absorption rate. The outcome does not support free and full interchangeability in chronic stable graft recipients of the two products studied, unless validated clinical and laboratory conversion protocols for each kind of organ transplantation are enforce. PMID- 12776807 TI - Anaesthesia for shock wave therapy in musculoskeletal disorders: a preliminary report. AB - The potential for using external applied energy to rectify or ameliorate musculoskeletal disorders has been explored for decades. A shock wave is a pressure disturbance: tissue effect is cavitation, producing microtrauma or microfracture and haematoma formation, inducing, as to date is thought, increase in vascularization, increased soft callus and faster enchondral ossification. Anaesthesiological interest in this field is focused in non-union or delayed osseous union, joint stiffness or osteochondrosis and femoral head necrosis in adults. Actually, because of the pain associated with high energy extracorporeal shock wave therapy on bones, anaesthesia is necessary, but, since almost all patients have no complaint after treatment, there is no need of postoperative analgesia. Therefore, short duration anaesthetic techniques and agents should be preferred. Loco-regional anaesthesia or general anaesthesia are both suitable to the purpose. Fifty patients have been treated nowadays in our Institution with shock wave therapy needing anaesthesia. 18 patients (36%) received general anaesthesia. Since patient's stay in hospital was expected to be short, short duration agents have been used, avoiding those causing unpleasent side effects, first emesis. We used Propofol or Remifentanil by continuous infusion, titrated to maintain stable haemodynamics and an appropriate level of anaesthesia. The short duration of action of Propofol depends on its rapid elimination, whereas Remifentanil undergoes rapid biotransformation to minimally active metabolites. 32 patients (64%) received regional anaesthesia. We avoided long acting agents or high concentration drugs. Spinal blocks have been performed with 0.5% hyperbaric bupivacaine; brachial plexus blocks, sciatic-femoral blocks and an epidural block have been performed with 0.5-1% xylocaine or 1% mepivacaine. Shock Wave Therapy has been done during a 3-day hospital stay. With suitable anaesthesiological treatment and preparation, almost all patients could be treated as outpatients or with an overnight hospital stay. PMID- 12776808 TI - Effect of epoetin on HO-1 mRNA level and plasma antioxidants in hemodialysis patients. AB - OBJECTIVE: Patients with renal failure and undergoing hemo- (HD) or peritoneal dialysis are under oxidative stress which is thought to contribute to the long term complications noted in this patient population. One effect of HD-induced oxidative stress is via red blood cell (RBC) membrane lipid peroxidation leading to RBC destruction and anemia. Interaction of this oxidative stress with epoetin (EPO) treatment to increase RBC number and Hb concentration remains unexplored. PATIENTS AND METHODS: This preliminary study used RT-PCR as well as colorimetric based assay approaches to evaluate the effect of EPO-alpha treatment on markers of oxidative stress in hemodialysis patients. Eighteen patients (12 males, 6 females, age range 45 - 68), were treated with EPO-alpha (Eprex) 50 UI/kg thrice weekly over an 8-month study period. Monocytes were isolated at baseline, then monthly thereafter, monocyte heme-oxygenase-1 (HO-1) and plasma Hb and antioxidant power (AOP) were determined. RESULTS AND CONCLUSIONS: Treatment with EPO increased Hb (9.4 +/- 0.7 g/dl to 10.9 +/- 0.5, mean +/- SD p < 0.001). In addition, both monocyte HO-1 mRNA (0.34 +/- 0.08 vs. 0.59 +/- 0.02 d.u. p < 0.001) and plasma AOP (1,379.8 +/- 175 micromol/l to 1,624 +/- 170, p < 0.04) increased. While AOP changes showed no correlation with other indices, increases in HO-1 and Hb were positively correlated using 2 different measures: delta Hb (peak Hb - baseline Hb) vs. delta HO-1 (peak HO-1 mRNA - baseline HO-1 mRNA) as well as delta Hb(5 months-baseline) vs. delta HO-1 (5 months - baseline) mRNA (r = 0.81, p < 0.001 and r = 0.76, p < 0.001; respectively). In conclusion, the increases upon EPO treatment of both HO-1 gene expression and plasma AOP as well as the significant correlation between delta Hb and delta HO-1 mRNA suggest that EPO treatment reduces oxidative stress via a combination of effects. These could potentially include effects on oxidative stress directly as well as effects on the levels and types of antioxidants present in plasma. PMID- 12776809 TI - Safety aspects of a coumarin-troxerutin combination regarding liver function in a double-blind placebo-controlled study. AB - OBJECTIVE: Coumarin is reported to elevate liver function tests (LFT) values. In a prospective, placebo-controlled, clinical trial, efficacy and safety of a coumarin-containing combination (SB-LOT) were evaluated in the treatment of chronic venous insufficiency. Here, we report on the drug safety of coumarin with special respect to liver reaction. METHODS: 114 patients were treated with SB-LOT (30 mg coumarin and 180 mg troxerutin t.i.d.) and 117 with placebo during a period of 16 weeks. LFT values (ALT, AST, AP and gamma-GT) were monitored at baseline, 4, 6, 8, 12 and 16 weeks of therapy. Adverse drug reactions were assessed regarding causality. Additionally, lymphocyte proliferation test was used to identify allergic reactions. Logistic regression analysis was performed to identify possible risk factors. RESULTS: No serious adverse drug reactions occurred. Elevations of LFT were assessed as biochemical abnormality. Specific clinical symptoms such as jaundice did not occur. Only 1 patient reported fatigue and exhaustion. Logistic regression estimated a basic risk for elevation of LFT of 4.9% under SB-LOT and 2.1% under placebo. Hepatitis in the history and diseases of the liver were identified as risk factors. CONCLUSION: This evaluation contributes to safety data of SB-LOT in man. LFT elevation is transient and the low risk of the SB-LOT therapy to increase LFT value can be limited when risk factors are considered. PMID- 12776810 TI - Pharmacokinetics of lithium in healthy volunteers after exposure to high altitude. AB - INTRODUCTION: Exposure of the human body to high altitude causes a number of physiological changes. In previous studies, we observed that these changes may alter the pharmacokinetics of drugs. The number of erythrocytes/mm3 increases both, after acute exposure to high altitude (HA), i.e. within 12 - 24 h after reaching high altitude (H), as well as in chronic exposure (HC) (> 10 months) to H. Also binding of drugs to biologic material may change with exposure to HA and/or HC. OBJECTIVE: Since lithium is transported into and out of erythrocytes and binds strongly to erythrocytes, but is not plasma protein-bound, we selected this drug as candidate for the present study. SUBJECTS, MATERIAL AND METHODS: Lithium carbonate 300 mg were administered orally to young healthy volunteers. One group residing at low altitude (Santiago, Chile, 600 m, group L), these same volunteers after 15 hours of exposure to high altitude (4,360 m, group HA), and volunteers living at high altitude for at least 10 months (group HC). RESULTS: We found a significant increase of both hematocrit and red blood cell count (RBC) after exposure to H, both, acute or chronic. Elimination half-life increased 64.1% in group HA and 111.4% in group HC in comparison to group L. We also found an increase in volume of distribution: + 18.9% in group HA, and + 35.8% in group HC when measured in plasma, and + 16.9% in group HA and + 18.8% in group HC when measured in whole blood. Lithium uptake by the erythrocytes increases: the value of 36.7 +/- 22.7% in Group L rose to 54.8 +/- 21.1% and to 54.6 +/- 24.2% in groups HA and HC, respectively. Total clearance decreases at high altitude, though the differences were significant only in group HC (37%). CONCLUSION: Results indicate that exposure to H produces alterations in the pharmacokinetics of lithium and that these variations may be clinically relevant. PMID- 12776811 TI - Trends in prescribing proton pump inhibitors in Taiwan: 1997 - 2000. AB - OBJECTIVE: The prescribing of proton pump inhibitors (PPIs) had increased greatly in recent years worldwide. Aim of our study was to analyze the utilization patterns of PPIs within the National Health Insurance program in Taiwan from 1997 - 2000. MATERIAL AND METHOD: The systemic sampling data-sets from the National Health Insurance Research Database served as data sources. Units of measurement for PPIs were numbers of prescription items and defined daily doses (DDDs). To estimate the proportion of the population treated daily with PPIs, numbers of DDDs per 1,000 inhabitants per day were calculated. In order to realize the role of PPIs in treating Helicobacter pylori-related disorders, we also analyzed various combined prescriptions of PPIs with amoxicillin, clarithromycin, metronidazole, tetracycline and bismuth. RESULTS: In the study period, PPI prescriptions increased nearly 2-fold at the outpatient sector and more than 3 fold at the inpatient sector. Men received more PPI prescriptions, as a whole, than women. Most PPIs were prescribed at the outpatient sector: 93.9% in 1997, 92.3% in 1998, 90.4% in 1999 and 87.3% in 2000. The numbers of DDDs per 1,000 inhabitants per day for all kinds of PPIs were 0.59, 0.78, 1.07, and 1.13 from 1997 - 2000, respectively. While the percentage of monotherapy increased from 63.6% in 1997 to 75.5% in 2000, the combination therapies did not experience a rapid and sustained growth. Among the combination therapies, PPI + amoxicillin + metronidazole and PPI + amoxicillin were popular in 1997 and 1998, but triple therapy with PPI + amoxicillin + clarithromycin predominated in 1999 and 2000. CONCLUSIONS: Despite increasing use, prescribing volumes of PPIs in Taiwan were far fewer than those in other developed countries. Treatment of Helicobacter pylori-related disorders in Taiwan followed universal standard. PMID- 12776812 TI - Henoch-Schonlein purpura associated with clarithromycin. Case report and review of literature. AB - OBJECTIVE: To report a case of Henoch-Schonlein purpura that appears to be related to the intake of clarithromycin for pharyngitis/tonsillitis. CASE SUMMARY: We describe a case of Henoch-Schonlein associated with clarithromycin therapy in a 48-year-old white man with no history of allergic drug reactions. Four days after starting therapy, he came to our hospital emergency room because of a non-pruritic palpable purpuric rash on the trunk and extremities and arthralgias involving elbows and knees. Administration of clarithromycin was suspended, in a few days, arthralgias and skin lesions quickly resolved. Three weeks later, the patient presented again with abdominal pain, dark-red urine and swelling of the legs. Urinalysis revealed proteinuria of 11 g/24 h and hematuria. A percutaneous renal biopsy showed a diffuse endocapillary proliferative glomerulonephritis with segmental areas of fibrinoid necrosis within glomeruli, on immunofluorescence study granular deposits of IgA and C3 were present in the mesangium and capillary walls. A diagnosis of HSP was made. We suspected that the causative agent might be clarithromycin since this was the only drug added before the cutaneous and renal condition appeared. CONCLUSIONS: Our case and the previous case suggest that HSP may represent a potential adverse effect of clarithromycin, clinicians should be alerted to this potentially severe side effect of such a widely used drug. In accordance with the data obtained and based on the Naranjo algorithm, the adverse reaction could be considered possible. PMID- 12776813 TI - Scaling or wider bioequivalence limits for highly variable drugs and for the special case of C(max). AB - OBJECTIVE: To illustrate that bioequivalence (BE) can be effectively evaluated for highly variable (HV) drugs and drug products and for the special case of C(max) by using average BE. To demonstrate that either scaling or wider regulatory limits need not result in large observed ratios of the geometric means (GMR) of the 2 drug products. METHODS: Two- and 4-period crossover BE investigations with 24 subjects were simulated. Variabilities of 15, 25 or 35% were assumed in special studies of C(max) and 40% in the general investigations of HV drugs. Acceptance of BE was analyzed in each study by various procedures and regulatory criteria. Under each condition, the percentage of simulated investigations accepting BE was recorded as the simulated GMR was gradually raised from 1.00. RESULTS: Scaled average BE for HV drugs (in both 2- and 4 period studies) and expanding limits for C(max) increased substantially, as expected, the proportion of investigations accepting BE. An additional secondary regulatory criterion constrained the simulated GMR to 1.25 and limited the possibility of large deviations between the mean metrics of the 2 formulations. Acceptance of BE by the composite regulatory expectation never exceeded the acceptances by the separate component criteria. CONCLUSIONS: The sample size required for the evaluation of BE for HV drugs and drug products can be substantially reduced by applying the approach of scaled average BE. The same conclusion is reached from the determination of BE for the C(max) metric by expanding the regulatory limits to 0.75 - 1.33 or even to 0.70 - 1.43. Concerns for observations of high GMR values can be eased by imposing constraints with a secondary regulatory criterion. PMID- 12776814 TI - Bioequivalence study of two brands of enalapril tablets after single oral administration to healthy volunteers. AB - OBJECTIVE: To compare the relative bioavailability and bioequivalence of 2 enalapril tablet formulations in healthy volunteers under fasting conditions. METHODS: An open-label, single-dose, randomized, two-period, crossover trial with a 1-week washout period in 24 healthy volunteers. The 2 enalapril 20 mg tablet formulations used were Antiprex (Elpen, Greece) as test and Renitec (Vianex, Greece) as reference preparation. Serial blood samples were collected at 19 points for 36 h. Plasma samples were analyzed for enalaprilat, the pharmacologically active metabolite of enalapril, by a validated GC/MS assay. Pharmacokinetic parameters, such as AUC(0-infinity), AUC(0-t), C(max) T(max), t1/2 and MRT were calculated from plasma concentrations for both formulations. Statistical comparisons (ANOVA and 90% confidence intervals) of AUC(0-infinity), AUC(0-t) and C(max) data were evaluated after logarithmic transformation, and differences of T(max) were tested non-parametrically. RESULTS: The parametric 90% confidence intervals of the geometric mean values of the test/reference ratios were 88.0 - 117.6% (point estimate: 101.8%) for AUC(0-infinity), 88.7 - 118.9% (point estimate: 102.7%) for AUC(0-t), and 91.0% - 123.4% (point estimate: 106.0%) for C(max) No statistically significant differences were found between the 2 preparations for T(max) t1/2 and MRT values. CONCLUSIONS: From the results of the present study, it is concluded that the test and reference tablet formulations of enalapril are bioequivalent for both the extent and the rate of absorption and therefore the 2 products can be considered to be interchangeable in clinical practice. PMID- 12776815 TI - Inhibition of return using discrimination of location. AB - The present study investigated "inhibition of return" which refers to increased response latency when the target in a location discrimination task appears in the same location on consecutive trials. Location discrimination tasks were performed by 28 university students (19-21 years of age) in two experiments. In Exp. 1 the task was to press a left or right button in response to a stimulus displayed on a computer screen. The first condition manipulated stimulus-response compatibility which resulted in changing the inhibition of return. Inhibition of return was stronger when the condition was stimulus-response incompatibility. This result shows that inhibition of return functions strongly under incompatible conditions. A second condition required performance of dual tasks in Exp. 2. Again, changes in inhibition of return were seen. These results show that inhibition of return functions strongly under both dual task and incompatible conditions. Inhibition of return was interpreted as functioning strongly when the task was more difficult. PMID- 12776816 TI - The plural self. AB - In a sample of 61 psychology students, unity in the subjective structure of the self was associated with having a Taoist orientation. PMID- 12776817 TI - A 3-year longitudinal study of self-concept and classroom behavior of grade 1 retained pupils. AB - The purpose of this study was to examine the relationship between repeating Grade 1, self-concept scores, and classroom behavior. In Canada, a large number of pupils repeat grades. The literature on repeating a grade suggests that this practice has no benefit on academic achievement or social behavior and practically none on self-concept. Data from a 3-yr. longitudinal study indicate that Grade 1 repetition has no clear relation with self-concept scores and classroom behavior, even when remedial teaching provided during or after the repeated grade is taken into account. It appears classroom behavior problems are related to other factors which existed prior to grade repetition. Furthermore, self-concept tends to be related to achievement rather than grade repetition. PMID- 12776818 TI - Perceptions of the importance of training, experience, and athletic talent for achievement in sports. AB - Ratings by 97 NCAA Division I athletes showed belief that successful athletes benefit more from training and experience and athletic talent than less successful athletes. PMID- 12776819 TI - Perceived standing position after reduction of foot-pressure sensation by cooling the sole. AB - We investigated the influence of the reduction of foot-pressure sensation by cooling the sole of the foot, at 1 degree C for 30 or 40 minutes, on the perception of standing position varied in the anteroposterior direction. The subjects were 16 healthy undergraduates. Firstly, for 4 of the subjects, cooling the sole of the foot decreased sensory information from the mechanoreceptors in the sole, by testing for an increase in the threshold for two-point discrepancy discrimination on the sole of the foot and for the disappearance of postural change with vibration to the sole. Next, the perception of standing position was measured by reproduction of a given standing reference position involving forward or backward leaning under both normal and cooled conditions of the feet. Standing position was varied in relation to the location of the center of foot pressure, defined as distance from the heel in percentage of the length of the foot. The reference positions, representing various locations of the center of foot pressure, were set at 10% increments from 20% to 80% of the length of the foot. With eyes closed, the subject first experienced the reference position and then attempted to reproduce it. The mean location of the center of foot pressure in the quiet standing posture was 45.7%. At the 40%, 50%, and 60% reference positions, those closest to quiet standing, absolute errors of reproduction were significantly larger than at other reference positions in both the normal and the cooled conditions. They were significantly larger in the cooled than in the normal condition. The 50% and 60% reference positions were reproduced significantly further forward in the cooled than in the normal condition. These results may be explained as due to an absence of marked changes in sensory information from both muscular activity and foot pressure when moving to reference positions close to the quiet standing posture. PMID- 12776820 TI - Cognitive correlates of laterality in mental retardation. AB - Correlations of various laterality score for 202 persons with cognitive test scores in the seventies of mental retardation are presented. PMID- 12776821 TI - Motivational climate and perceived competence in anxiety and tennis performance. AB - This study was done to verify a prediction in achievement goal-orientation theory in a Korean physical education setting. It was hypothesized that the interactions of situationally induced motivational climates (Task and Ego) and perceived competence would differentially influence anxiety and performance in tennis classes. 60 undergraduate men were assigned to a Task-involving and an Ego involving program in tennis classes for 6 wk. Their perceptions of motivation climates and tennis competence were measured after the first 3 wk., and anxiety responses (cognitive and somatic) and tennis skill performance were evaluated every week over the second 3 wk. of the class. Analysis showed in Task-involving condition students decreased anxiety responses and increased tennis performance, whereas Ego-involving students who had low perception of their tennis competence maintained their anxiety responses and decreased their tennis performance. These results support the hypotheses and the predictions of goal-orientation theory and suggested practical application of the goal-orientation theory in Korea could be explored. PMID- 12776822 TI - Use of self-reports of physical fitness as substitutes for performance-based measures of physical fitness in older adults. AB - The purpose of the present study was to evaluate the association between self reported physical fitness and performance-based measures of physical fitness in older adults. The specific components of physical fitness evaluated included aerobic endurance, muscular strength, and flexibility. Adults (25 men and 47 women) ranging in age from 56 to 92 years (M age=75 yr.) were recruited from the local community. Generally, the associations between self-reported and performance-based measures of physical fitness were low to moderate (r = 30-.01). Based on these findings, self-reports of physical fitness should not be used as substitutes for performance-based measures of physical fitness in older adults. Furthermore, present findings suggest that older adults, when asked to rate subcomponents of physical fitness, may not do so but rather evaluate a more general concept of physical fitness with aerobic endurance as the dominant factor. PMID- 12776823 TI - Seasonality of suicide in eastern Europe: a comment on "Evidence for lack of change in seasonality of & suicide from Timis County, Romania". AB - The seasonality of suicide in Hungary declined from the 1980s to the 1990s, contradicting the recent conclusion of Voracek, et al. PMID- 12776824 TI - Influence of instruction on velocity and accuracy of overarm throwing. AB - This study investigated the influence of instruction on the ball velocity and accuracy of a goal-directed overarm throw. 9 experienced Norwegian male team handball players executed a throwing task randomly seven times under 5 different instructions varying from emphasizing speed to accuracy. When instructions increasingly emphasized accuracy, velocity decreased. However, accuracy did not improve when subjects were instructed to focus on it. A possible explanation for this finding could lie in the specific subject group. The subjects were highly experienced team handball players with an average of 12 years in the sport. Thus, the accuracy they demonstrated at high velocity might actually be already extremely high and difficult to improve upon when reducing throwing velocity. Further, the velocity of throwing when instruction emphasized accuracy was approximately 85% of the maximal velocity, indicating that experienced team handball players are trained to throw accurately at relatively high velocity. PMID- 12776825 TI - Re-examining the factor structure of the California Critical Thinking Disposition Inventory. AB - This study re-examined the factor structure of the California Critical Thinking Disposition Inventory developed by Facione and colleagues. Analysis showed that the four factors had some cross-validity with four factors in Walsh and Hardy's 1997 study. The results suggest the possibility of developing a more efficient short version of the inventory and calls for further improvement. PMID- 12776826 TI - Variations in mood state as a function of a football season. AB - Scores on the Profile of Mood States were collected from football players at a small university each week prior to a Saturday football contest. The scores were analyzed to assess whether mood varied as a function of the outcome of the previous week's game. Although the subjects' overall mood state was not influenced by the outcome of the games, scores on Tension, Vigor, Fatigue, and Confusion varied. Fatigue and Confusion were related to the outcomes of the games and gave partial support to the hypothesis that the players' mood would vary according to the outcome of the games played. PMID- 12776827 TI - Perceptual judgements of others' tasting experiences: are they enjoying their food? AB - Motivated by previous work suggesting that infants make stereotypic facial reactions to different tastes, we assessed communicative signals that might enable an adult to interpret the strength, taste, and hedonic value of a liquid flavour another adult is consuming. Four subjects (tasters) were overtly videoed consuming drinks that varied in strength (low, medium, and high concentrations), taste (sweet, sour, bitter, and salty), and hedonic value (taster-rated enjoyment). 26 observers assessed the strength, taste, and taster's enjoyment of the drink from video clips of the tasters. Observers perceived the hedonic value of the drinks to the taster and the drinks' strength based on the tasters' reactions but were generally poor at assessing the specific taste of the drink. For all tastes except the bitter ones, observers might have based their judgements of taste on how much the taster appeared to enjoy the drink. These findings are discussed in terms of communication of food's value. PMID- 12776828 TI - A cognitive-behavioural analysis of mental toughness in national rugby league football teams. AB - This study examined the relations between demographic characteristics of rugby players and selected aspects of psychological performance in rugby league football. Mental toughness was assessed using Psychological Performance Inventory and Hardiness on the Personal Views Survey III-R. Participants (N=70) were international rugby league footballers representing four teams (Wales, France, Ireland, England) in the 2000 Rugby League World Cup. Participants completed the questionnaires in training camp. Welsh-nationality players had a significantly higher mean score on two of the hardiness subscales. Hardiness measures displayed the greatest and most frequently statistically significant differences. The findings concur with previous work indicating superior hardiness is related to improved performance in sports. PMID- 12776829 TI - Race and perception of acceptance in nontraditional sports. AB - Analysis of responses from 632 respondents to a survey representing 4 military and 4 college locations in the southeastern USA indicated that race was a significant factor in perceptions of "welcomeness" or acceptance for seven of 10 non-traditional sports (70%). These significant differences persisted across sex, income, education, and location categories. When "welcome" scale scores were ranked, however, a strong association in ranking (Spearman rho=.66, p<.05) was found among racial groups. While it appears that many African Americans rated being much less welcome in nontraditional sports than Euro-Americans, both racial groups were likely to agree on which nontraditional sports would be more "welcoming" to new participants. PMID- 12776830 TI - Interest and attention in facial recognition. AB - When applied to facial recognition, the levels of processing paradigm has yielded consistent results: faces processed in deep conditions are recognized better than faces processed under shallow conditions. However, there are multiple explanations for this occurrence. The own-race advantage in facial recognition, the tendency to recognize faces from one's own race better than faces from another race, is also consistently shown but not clearly explained. This study was designed to test the hypothesis that the levels of processing findings in facial recognition are a result of interest and attention, not differences in processing. This hypothesis was tested for both own and other faces with 105 Caucasian general psychology students. Levels of processing was manipulated as a between-subjects variable; students were asked to answer one of four types of study questions, e.g., "deep" or "shallow" processing questions, while viewing the study faces. Students' recognition of a subset of previously presented Caucasian and African-American faces from a test-set with an equal number of distractor faces was tested. They indicated their interest in and attention to the task. The typical levels of processing effect was observed with better recognition performance in the deep conditions than in the shallow conditions for both own- and other-race faces. The typical own-race advantage was also observed regardless of level of processing condition. For both own- and other-race faces, level of processing explained a significant portion of the recognition variance above and beyond what was explained by interest in and attention to the task. PMID- 12776831 TI - The inappropriate smile and zygomatic muscle activity. AB - Long-term recording of activity of the zygomatic muscle, the most important mimic muscle involved in smiling, was performed in 22 disorganized type schizophrenic patients with inappropriate smiles and 15 normal subjects in two separate experiments. During inappropriate smiles, the zygomatic muscle exhibited waxing and waning bursts of activity with an amplitude of 75 to 120 microV, which could not be distinguished from the activity observed during usual smiling in normal subjects. However, the duration of such activity tended to be longer when compared to that in usual smiling by normal subjects as well as by schizophrenic patients. In contrast to usual smiles, inappropriate smiles decreased with personal contact. When asked about their thoughts during smiling shortly after inappropriate smiles, more patients reported that they thought of nothing at all or something not necessarily pleasant rather than something pleasant that would be expected to induce smiling. PMID- 12776832 TI - Intensity and color of language in attitude change and emotion. AB - This study examined whether messages using or not using emotionally intense language combined with certain colors, i.e., red, white, or blue, to effect attitude change. Emotionally intense messages were more positively associated with attitude change than were those with low emotional intensity, but no interaction effects or main effects for color were found. PMID- 12776833 TI - Cross-script and within-script priming in alcoholic Korsakoff patients. AB - In two experiments, alcoholic Korsakoff patients and control subjects studied a list of Japanese nouns written in either Hiragana or Kanji script. Word-fragment completion and recognition tests were then administered in Hiragana. When the writing script was changed between study and test phases, repetition priming in word-fragment completion was significantly attenuated but was still reliable against baseline performance. This was confirmed for both Korsakoff patients and control subjects. In contrast, the script change had little effect on recognition memory, which was severely impaired in Korsakoff patients. The results suggest that repetition priming is mediated by two different implicit processes, one that is script-specific and the other that is assumed to operate at a more abstract level. PMID- 12776834 TI - Cognitive strategy and ability in endurance activities. AB - The purpose of this study was to ascertain whether elite athletes utilize associative strategies and, on the other hand, if nonelite athletes prefer to use dissociative strategies. 60 athletes were interviewed and the Schomer's 1986 method of measurement was used to measure association and dissociation. Analysis showed no significant differences in the total associative and dissociative scores among the three different abilities. However, a significant difference was found concerning the use of dissociative strategy between men and women. PMID- 12776835 TI - Changes across age groups in self-choice elaboration and incidental memory. AB - This study investigated differences in the self-choice elaboration and an experimenter-provided elaboration on incidental memory of 7- to 12-yr.-olds. In a self-choice elaboration condition 34 second and 25 sixth graders were asked to choose one of the two sentence frames into which each target could fit more congruously, whereas in an experimenter-provided elaboration they were asked to judge the congruity of each target to each frame. In free recall, sixth graders recalled targets in bizarre sentence frames better than second graders for self choice elaboration condition. An age difference was not found for the experimenter-provided elaboration. In cued recall self-choice elaboration led to better performance of sixth graders for recalling targets than an experimenter provided elaboration in both bizarre and common sentence frames. However, the different types of elaboration did not alter the recall of second graders. These results were interpreted as showing that the effectiveness of a self-choice elaboration depends on the subjects' age and the type of sentence. PMID- 12776836 TI - Expressive writing disorders: an analysis of theoretical views. AB - Three prominent theories of writing were found in a literature review of Psychlit, educational journals and Medline, and a fourth global theory relating writing to intact cerebral integrity was also hypothesized. These theories were then tested by attempting to predict writing scores among a sample of 44 individuals with academic difficulties. Significant support was found for a memory and attention model, the dysgraphia model, and an abstract sequential analysis model. Surprisingly, measures of the overall cortical integrity were not significantly related to writing behavior in this sample, but this finding may reflect excluding brain damaged subjects in this study. Also, the abstract sequential analysis model appeared to function differently than expected. The results indicate it is important to assess the cognitive contributions of writing difficulties to guide remediation. Larger scale research on the sources of writing disorders is also recommended. PMID- 12776837 TI - Effects of classroom noise on students and teachers in Greece. AB - Measurements inside 32 occupied high-school classrooms during the courses showed that the measured long-term equivalent noise levels are high. A significant percentage of students and teachers found these levels unacceptable. The observed noise levels depend on the time period of the course, the age, and the number of the students in each classroom. The large reverberation time and the high background noise are responsible for the measured high noise level. PMID- 12776838 TI - English letter frequencies and their applications: part I. AB - This article presents data regarding the relative frequencies of single letters in English, organized by word length and letter position. Derived from a parsimonious sample of English word use patterns presented by Whissell in 1998, the data accurately represent letter frequencies found in modern English. These data provide a resource for various applications, including reading research and practice. PMID- 12776839 TI - Perceptibility of large and sequential changes in somatosensory information during leaning forward and backward when standing. AB - 11 healthy young men served as subjects in two experiments on perceptibility of (1) large changes in foot pressure and muscle activity induced by body leaning and (2) sequential changes in pressure at the first toe and the head of the first metatarsalis when leaning forward. The effects of reduced sensitivity on that perceptibility were also studied by repeating the experiments while cooling localized plantar areas of the sole (the head of the first metatarsalis, the first toe, and the heel). Under the normal (noncooled) condition, all subjects accurately perceived maximum pressure at the head of the first metatarsalis, but most subjects misperceived the second large increase in pressure at the first toe and in muscle activity as the first large increase. Under the cooling condition, localized cooling did not affect the perceptibility of maximum pressure at the head of the first metatarsalis or the activity in the tibialis anterior, but the perceptibility of pressure at the first toe and activity of the abductor hallucis were reduced. There were individual differences in perceptibility of activity of the rectus femoris when the heel was cooled. Perceptibility of sequential changes in the pressure was affected differently by the localized cooling of each region. Given these findings, we discussed the role and interrelatedness of pressure sensation in perceiving large and sequential changes in somatosensory information while standing and leaning forward and backward. PMID- 12776840 TI - Practice and incentive motivation in recognition of inverted faces. AB - In each of three experiments, participants received successive daily practice sessions on the task of recognizing inverted faces. In all practice sessions, an initial study series of 25 inverted faces was followed immediately by a test series of 17 pairs of inverted faces. Each test pair comprised a face from the study series and a new face. Completely new sets of faces were used in each session. Recognition of inverted faces did not improve across sessions in Exp. 1 but did improve in Exps. 2 and 3. Unlike Exp. 1, Exps. 2 and 3 employed an explicit incentive for improved performance. These results show that sufficiently motivated participants can become quite proficient at recognizing inverted faces. Implications of the results for the role of expertise at recognition in producing the inversion effect are discussed. PMID- 12776841 TI - Developmental differences in children's ballistic aiming movements of the arm. AB - An experiment was conducted to examine the change in the relation between programming and "on-line" correction as a developmental explanation of children's arm movement performance. Each of 54 children in three age groups (5, 8, and 10 yr.) completed two types of rapid aiming arm movements in the longitudinal plane on the surface of a digitizer. Percent primary submovements and timing variability were dependent variables. Analysis suggested that the 5-yr.-olds used "on-line" monitoring during the arm movement and did not perform the movement sequence as a functional unit. Compared with 8- and 10-yr.-olds, the 5-yr.-olds planned a smaller portion of movements, executed the arm movements with more variability in time to peak velocity. The 8- and 10-yr.-olds appeared to plan their movements and execute the sequence as a unit. The developmental implications were discussed. PMID- 12776842 TI - Test of the effect of attention on judged length of a line. AB - Studies to assess experimentally whether attention affects the judged length of a line have produced discordant results. This paper reports the results of a test designed to avoid factors that were not controlled in previous studies. Stimuli were either two vertical lines or two horizontal lines of equal physical length presented briefly on opposite sides of a fixation cross. Subjects were asked to direct their attention to one line when the arm of the cross pointing to the line changed in luminance. This arm was used either as a precue or as a postcue. Subjects judged lines to be longer when a precue preceded the line than when a postcue followed the line. PMID- 12776843 TI - Effects of a carbohydrate-restricted diet on affective responses to acute exercise among physically active participants. AB - Currently, the carbohydrate-restricted diet is very popular. Atkins' book, Dr. Atkins' Diet Revolution, has sold millions in its more than 25 years of existence. His book promotes the carbohydrate-restricted diet, which focuses on the consumption of proteins and fats as primary calorie and energy sources, while severely restricting carbohydrates. However, when carbohydrates are restricted from the diet, the body's primary energy source is reduced considerably. Therefore, the purpose of this study was to compare the psychological responses to exercise of individuals when on a carbohydrate restrictive diet and when on a noncarbohydrate restrictive diet. For this study, 17 participants practiced a noncarbohydrate-restricted diet for three weeks and the carbohydrate-restricted diet for three weeks, while maintaining previous exercise habits. After each exercise session, the participants completed the Physical Activity Affect Scale, which measures Positive Affect, Negative Affect, Tranquility, and Fatigue. Simple one-way analyses of variance indicated significant treatment differences (ps<.05) relative to Negative Affect, Positive Affect, and Fatigue. The results of the study indicate as predicted, that, when a person restricts carbohydrates from the diet, he will experience more fatigue, more negative affect, and less positive affect in response to exercise than those individuals who are not restricting carbohydrates. PMID- 12776844 TI - Hostility disturbs learning. AB - Many studies have shown that there is a strong correlation between hostility and coronary artery disease; however, the pathogenic mechanisms by which hostility causes coronary artery disease have not been identified. Several studies have shown that hostility is associated with increased cardiovascular reactivity to mental stress. Sloan and colleagues used mental arithmetic and the Stroop Color Word Task as psychological stressors and suggested that hostility is associated with diminished cardiac vagal control. It is supposed that the diminished cardiac vagal control results in uncontrollability of increased heart rate under stressful conditions so performance on mental stress tasks is poor. However, performance was not analyzed on the Stroop Color-Word Task. If hostility influences the autonomic nervous system, the performance of this mental stress task may also differ according to extent of hostility. In the present study, whether hostility disturbed performance of a mental stress task and the practice on it was examined. Subjects completed the Cook-Medley Hostility Scale and were divided into three groups (High, Middle, and Low) by their total scores and three subscales (Cynicism, Hostile Affect, and Aggressive Responding). They also completed the Stroop Color-Word Task. Analysis showed practice by High and Middle scoring groups on Aggressive Responding had a significantly smaller effect than that by Low scoring groups. The pathogenic mechanisms by which hostility may underlie coronary artery disease were discussed. PMID- 12776845 TI - Ambient sensory conditions: modification of receptive speech deficits in left side stroke patients using bright light. AB - Previous research has demonstrated the potential for the use of ambient sensory conditions to improve cognitive functioning. Both light and sound have been shown to improve task performance in various populations including children, younger adults, and elderly participants; however, these cognitive gains may possibly be offset by detrimental cardiovascular reactivity in elderly persons. We now have some evidence for ambient sensory conditions affecting cognitive function following left- or right-side cerebrovascular accidents. PMID- 12776846 TI - Role of perceptual and motor abilities in instep-kicking performance of young soccer players. AB - The present study made a dynamic analysis of the ground reaction forces developed on the supporting foot during instep kicking to investigate the relation between specific perceptual and motor abilities and the performance of this skill. 45 young soccer players (11-13 years of age) participated in a series of laboratory tests assessing simple, choice, and discrimination reaction time, sustained attention, depth perception, and sense of kinesthesis. Kicking performance measured by the amount of impulse (calculated as the integral of force) developed on the supporting foot during kicking. There was a significant correlation of the kicking impulse with choice reaction time (r = -.54) and attention reaction time (r = -.41). Stepwise regression analysis indicated that choice reaction time accounted for 29% of the variation in the anterior/posterior kicking impulse and 16.4% of the variation in the medio/lateral kicking impulse. The significant relation between kicking impulse and measures concerning speed of information processing suggests that processes associated with fast response selection may play an important role in instep-kicking performance. These findings can provide useful information for designing of training schemes and testing protocols. PMID- 12776847 TI - Development of a Spanish version of the Participation Motivation Inventory for young competitive swimmers. AB - The present study examined the validity of a Spanish version of the Participation Motivation Inventory, modified for swimming by Gould, Feltz, and Weiss in 1985, and investigated whether young swimmers differing in sex and age vary in their objectives for participation. Swimmers (204 boys and 224 girls, ranging in age from 8 to 22 years) were administered the inventory. Factor analysis with varimax rotation identified seven factors fairly consistent with previous research. Differences were found across age and sex. Females placed greater emphasis than males on Friendship and Fun and indicated lower importance rating to Status. Younger children (8-10 years) rated Fun/Friendship, Competition/Skills, Significant Others, and Status significantly higher in importance than did the other age groups. PMID- 12776848 TI - A brief (one-hour) quantitative neuropsychological assessment with three performance-based tests: strong concordance with proficiency scores for a more extensive test battery. AB - Raw scores for each of several dozens of traditional and more recently developed neuropsychological tests were correlated with an impairment index composed of all of these scores from the records of 162 patients who had been assessed following impacts of substantial mechanical energies. A score of either less than 20 correct binaural responses for a dichotic word listening task, more than 99 sec. for Trails B, and more than 3.8 min. to complete the Tactual Performance Test with both hands correctly classified 85% of patients whose z scores were less than -1.0 (below average) or -1.0 or above (average) for a composite neurocognitive index. The results suggest that these three tests, administrable within about one hour, may be employed as a more objective criterion rather than "clinical impressions" for discerning if patients require more extensive neuropsychological testing. PMID- 12776849 TI - Relations between perceptual and physiological response during incremental exercise followed by an extended bout of submaximal exercise on a cycle ergometer. AB - The purpose of this study was to examine the relations of ratings of perceived exertion (RPE) of the legs, chest, and overall body with physiological responses (heart rate and oxygen uptake) both during incremental cycling exercise and the recovery stage (submaximal light exercise after total exhaustion). Subjects were 10 healthy university males ages 18 to 23 years (M age=20.5 yr., SD=1.4 yr.) who performed incremental cycling exercise until exhaustion after 1-min. rest and unloaded cycling for 2 min. They then continued to exercise at a constant load of 30 Watts (used for cooling down; recovery stage) for a total of 25 min. Oxygen uptake and heart rate were measured, and three types of RPE were done; Respiratory (chest; RPE-R), Peripheral (legs; RPE-P), and Overall (overall body; RPE-O) during the exercise and recovery stage. All variables during exercise and RPE-R and RPE-P during recovery stage showed significant linear changes. RPE-O and physiological exercise intensity (oxygen uptake and heart rate) in the recovery stage showed significant curvilinear changes (quadratic). RPE-P were significantly higher than RPE-R both during exercise and the recovery stage and the variables highly correlated (r > or = .88, p < .05). At the point of exhaustion, RPE-P and RPE-O almost reached a peak, but RPE-R did not. In the exercise period until exhaustion, the regression coefficient of RPE-R (.38) was significantly lower than that of RPE-P (.56) and RPE-O (.50), and RPE-R increased according to an increase of the incremental load, but the amount was significantly lower than those of RPE-P and RPE-O. In the recovery stage after exhaustion, the regression coefficient of RPE-O (-1.35) was significantly greater than that of RPE-P (-1.07). A decrease in RPE-O corresponded to a decrease in heart rate and oxygen uptake, but RPE-P did not, and the recovery of RPE-P tended to be late. The results suggest that relations for the physiological responses of heart rate, oxygen uptake, and RPE, and between each RPE in the recovery stage differed from those during exercise until exhaustion. PMID- 12776850 TI - Influence of drafting during swimming on ratings of perceived exertion during a swim-to-cycle transition in well-trained triathletes. AB - After the swim to cycle transition of a triathlon, perceived exertion (RPE) during cycling was higher than during a single cycling bout for 8 well-trained triathletes, but swimming in a drafting position led to lower RPE responses and energy cost of cycling than swimming alone. PMID- 12776851 TI - Strength-based assessment of African-American adolescents with behavioral disorders. AB - The study examined consistencies and differences between 60 parents and their adolescent children with behavioral problems when rating the adolescents' strengths. The parents and teenagers agreed on most of the strength categories of the Behavior and Emotional Rating Scale. However, caretakers rated the adolescents as more involved in family life, while the adolescents rated themselves as more involved in school activities. PMID- 12776852 TI - Rats' preferences for an analgesic compared to water: an alternative to "killing the rat so it does not suffer". AB - A common policy in research institutions is to kill rats when they display chronic disabilities or recurrent injuries. These guidelines appear to be derived from an oxymoron that "it's better for a rat to be killed so it does not suffer pain" and from untested assumptions that rats cannot control "pain." In a two bottle paradigm, 10 rats with a history of brain damage following status epilepticus from a single systemic injection of lithium and pilocarpine were given options to consume freely either tap water or 1 mg/cc of acetaminophen in tap water. During periods of fresh lesions due to persistent gnawing or acute injuries associated with tonic-clonic convulsions, the rats consumed 3 to 10 times the fluid from the bottles containing acetaminophen (equivalent to 5 to 10 extra-strength Tylenol tablets per day for a 70-kg person) relative to periods when no lesions or old lesions were present. These results suggest that rats with chronic injuries sufficient to be terminated according to Animal Care guidelines may be capable of reducing the aversive physiological conditions associated with tissue damage by selecting analgesic treatments. PMID- 12776853 TI - Lack of association between negative priming and Necker Cube reversals. AB - 50 undergraduates were tested on two tasks presented in a counterbalanced fashion: their ability to inhibit irrelevant information measured on a negative priming task involving letter case information and on the number of Necker Cube reversals produced in a specific time period. Since number of reversals presumably reflects a person's ability to see various interpretations of the Necker Cube, this assumes that multiple interpretations are available to a subject and no one interpretation dominates over another. If true, then more reversals could indicate an ability to inhibit irrelevant information. That is, observing one reversal at the expense of another reversal indicates a subject can adequately inhibit these various interpretations successfully, with more reversals indicating a higher ability to inhibit information successfully. Although it was predicted that negative priming performance would be related to the number of Necker Cube reversals, this was not the case (r = -.07). PMID- 12776854 TI - Assessing handwriting speed of children biliterate in English and Chinese. AB - The assessment of multiliterate handwriting performance is rarely reported despite increased globalization. The present study describes the psychometric properties of a handwriting speed test developed for children who are biliterate in English and Chinese. This included interrater reliability, test-retest reliability, interitem correlation, construct validity, and concurrent validity. The test's reliabilities between two raters and over a 1-wk. interval were high with ICCs ranging from .89 to .99. Interitem correlation between the English and Chinese items was .87. The presence of age trends but not sex differences was a positive indicator of the test's validity. Correlations of .91 and 1.00 between the Chinese and the English items of the Handwriting Assessment Tool with the Chinese Handwriting Speed Test and Handwriting Speed Test, respectively, provided evidence of concurrent validity. These preliminary results showed the Handwriting Assessment Tool is reliable and is a potentially useful handwriting test for children biliterate in English and Chinese. The feasibility of assessing biliterate handwriting speed performance with the same set of scoring criteria for different writing systems was supported. PMID- 12776855 TI - Interaction of prime and target in the subliminal affective priming effect. AB - It has been found that an emotional stimulus such as a facial expression presented subliminally can affect subsequent information processing and behavior, usually by shifting evaluation of a subsequent stimulus to a valence congruent with the previous stimulus. This phenomenon is called subliminal affective priming. The present study was conducted to replicate and expand previous findings by investigating interaction of primes and targets in the affective priming effect. Two conditions were used. Prime (subliminal presentation 35 msec.) of an angry face of a woman and a No Prime control condition. Just after presentation of the prime, an ambiguous angry face or an emotionally neutral face was presented above the threshold of awareness (500 msec.). 12 female undergraduate women judged categories of facial expressions (Anger, Neutral, or Happiness) for the target faces. Analysis indicated that the Anger primes significantly facilitated judgment of anger for the ambiguous angry faces; however, the priming effect of the Anger primes was not observed for neutral faces. Consequently, the present finding suggested that a subliminal affective priming effect should be more prominent when affective valence of primes and targets is congruent. PMID- 12776856 TI - Heart rate turbulence after ventricular and atrial premature beats in subjects without structural heart disease. AB - INTRODUCTION: Studies assessing heart rate (HR) behavior after premature beats have focused on HR responses to ventricular premature beats (VBPs), but there is less information of HR behavior after atrial premature beats (APBs). METHODS AND RESULTS: HR turbulence after VPBs and APBs was first measured in response to ambient APBs and VPBs occurring during 24-hour ambulatory ECG recordings in 29 subjects without structural heart disease, and in response to programmed atrial (AE) and ventricular extrastimuli (VE) in 6 subjects undergoing electrophysiologic (EP) examination. Turbulence onset (TO) was more negative ( 2.3 +/- 3.2% vs -0.9 +/- 2.8%, P < 0.01) and turbulence slope (TS) was steeper (11 +/- 11 vs 5.1 +/- 4.1 msec/R-R interval, P < 0.05) after VPBs than APBs. Compared to VPBs, the acceleration of HR after APBs was delayed by one beat, and APBs were associated with a short R-R interval preceding the APB, resulting in a blunted TO. Studies of patients undergoing an EP test confirmed the one-beat delay of HR acceleration and the blunted TO after programmed AE compared to VE (P < 0.05). TO and TS after VPBs were related to baroreflex sensitivity. TO also was related to 24-hour standard deviation of N-N intervals (SDNN). However, the TO or TS following APBs was not related to either SDNN or baroreflex sensitivity. CONCLUSION: HR behavior is different in response to APBs and VBPs among subjects without structural heart disease. Different definitions and calculation formulas should be used in the analysis of HR turbulence after APBs and VPBs. PMID- 12776857 TI - Heart rate turbulence: explorations of an emerging risk factor. PMID- 12776858 TI - Natural history of Brugada syndrome: the prognostic value of programmed electrical stimulation of the heart. AB - INTRODUCTION: The prognostic value of electrophysiologic studies in individuals with the syndrome of right bundle branch block and ST segment elevation in precordial leads V1 to V3 (Brugada syndrome) remains controversial. Our previous data from 252 individuals with the syndrome suggested that programmed ventricular stimulation had a good overall accuracy to predict events. However, studies from independent investigators questioned our results. We report here the largest population with Brugada syndrome ever studied by programmed electrical stimulation of the heart. METHODS AND RESULTS: Four hundred forty-three individuals with an ECG diagnostic of Brugada syndrome were studied by programmed electrical stimulation of the heart. The diagnosis was made because of the classic ECG showing a coved-type ST segment elevation in precordial leads V1 to V3. Of the 443 individuals, 180 had developed spontaneous symptoms (syncope or aborted sudden cardiac death) and 263 were asymptomatic at the time the diagnosis was made. The ventricular stimulation protocol included a minimum of two basic pacing cycle lengths with two ventricular premature beats from the right ventricular apex. A sustained ventricular arrhythmia was induced in 217 cases (49%). Symptomatic patients were more frequently inducible [126/180 (70%)] than asymptomatic individuals [91/263 (34%); P = 0.0001]. Males were more frequently inducible than females (54% vs 32%, P < 0.0001). Inducible individuals had a longer HV interval than noninducible patients (50 +/- 12 msec vs 46 +/- 10 msec, P < 0.002). HV interval and number of premature beats needed to induce VF were not related to outcome. Inducibility was statistically a powerful predictor of arrhythmic events during follow-up. Sixty of 217 inducible patients (28%) had spontaneous ventricular fibrillation compared with 5 of 221 noninducible patients (2%; P = 0.0001). CONCLUSION: Inducibility of sustained ventricular arrhythmias during programmed ventricular stimulation of the heart is a good predictor of outcome in Brugada syndrome. PMID- 12776859 TI - Inducible ventricular fibrillation in the Brugada syndrome: diagnostic and prognostic implications. PMID- 12776860 TI - Change in the retrograde atrial activation sequence following radiofrequency modification of the atrioventricular node: implications for the electrophysiologic circuit of a variant of atrioventricular nodal reentrant tachycardia. AB - INTRODUCTION: Despite the great success in treating AV nodal reentrant tachycardia (AVNRT) with radiofrequency modification of the AV node, the dimensions of the electrophysiologic circuit of this arrhythmia remain unclear, and simple models fail to explain all tachycardia-related phenomena. METHODS AND RESULTS: We describe three unusual cases of supraventricular tachycardia (SVT). In all three cases, retrograde atrial activation during ventricular pacing or during SVT manifested local left atrial electrograms recorded from the coronary sinus preceding the septal atrial electrograms (eccentric activation), with earliest atrial activity at the lateral or posterolateral mitral annulus. Electrophysiologic maneuvers and observations were consistent with AVNRT as the mechanism in each case. In all cases, radiofrequency modification of the AV node eliminated inducible SVT and abolished dual pathway AV nodal physiology. The retrograde atrial activation sequence during ventricular pacing changed after ablation in each case, with septal atrial electrograms preceding left atrial electrograms recorded from the coronary sinus (concentric activation). CONCLUSION: The observations in these cases cannot be explained by the traditional model of slow, fast, and intermediate AV nodal pathways. A model incorporating a circuit close to the AV node with left atrial and coronary sinus connections is proposed. PMID- 12776861 TI - A simple method to identify sleep apnea using Holter recordings. AB - INTRODUCTION: Despite its severe cardiovascular and other consequences, sleep apnea syndrome frequently is undiagnosed. Because apneas result in repeated autonomic arousals associated with cyclic variations in heart rate (CVHR), we hypothesized that sleep apnea syndrome could be identified from simple HR tachograms (graphs of HR vs time) derived from ECG monitoring. METHODS AND RESULTS: HR tachograms were generated from 57 digitized ECGs (46 clinical patients undergoing diagnostic studies and 11 research subjects) obtained during overnight polysomnography. Thirty-three had significant sleep apnea syndrome (apnea-hypopnea index > or = 15). Eight patients had simultaneous Holter recordings during sleep studies (3 with digitized ECGs and 5 with paper ECGs). Duration of CVHR on tachograms was determined. CVHR patterns were characterized as high amplitude (HR changes > or = 20 beats/min per cycle) versus lower amplitude (6-19 beats/min per cycle); or regular (in frequency, amplitude, and morphology) versus irregular. Tachograms were classified as having visible HR changes versus not visible (flat). Twenty-four studies proved to be split-night, so CVHR was quantified for the first 3 hours of each study only. When subjects were dichotomized into shorter (< 20%, < 36 min) and longer (> or = 20%) durations of CVHR, longer CVHR had a positive predictive accuracy of 86% for significant sleep apnea syndrome and 100% for abnormal sleep. When flat tachograms were excluded, negative predictive accuracy for shorter CVHR was 100%. All patients (N = 13) with > 36 min high-amplitude CVHR had significant obstructive sleep apnea. All predictions from Holter-only data were concordant with clinical diagnoses. CONCLUSION: HR tachogram patterns derived from ambulatory ECGs provide a simple method for identifying sleep apnea syndrome and other sleep disturbances in patients without major autonomic dysfunction. PMID- 12776862 TI - Ambulatory electrocardiographic monitoring to screen for sleep apnea: is it really that simple? PMID- 12776863 TI - Magnetic resonance imaging findings in patients meeting task force criteria for arrhythmogenic right ventricular dysplasia. AB - INTRODUCTION: Magnet resonance imaging (MRI) findings in patients meeting Task Force criteria for the diagnosis of arrhythmogenic right ventricular dysplasia (ARVD) have not been systematically described. We report qualitative and quantitative MRI findings in ARVD using state-of-the-art MRI. METHODS AND RESULTS: MRI was performed on 12 patients with ARVD who were prospectively diagnosed using the Task Force criteria. The imaging protocol included breath hold double inversion recovery spin-echo and gradient-echo images. Ventricular volumes and dimensions were compared to 10 age- and sex-matched normal volunteers. High intramyocardial T1 signal similar to fat signal was observed in 9 (75%) of the 12 patients and in none of the controls. Right ventricular (RV) hypertrophy was seen in 5 (42%) patients, trabecular disarray in 7 (59%), and wall thinning in 3 (25%). Both the RV end-diastolic diameter and the outflow tract area were significantly higher in ARVD patients compared to controls (51.2 vs 43.2 mm, P < 0.01; and 14.5 vs 9.3 cm2, P < 0.01, respectively). ARVD patients had a higher RV end-diastolic volume index and lower RV ejection fraction compared with controls (127.4 vs 87.5, P < 0.01; and 41.6% vs 57%, P < 0.01, respectively). CONCLUSION: High intramyocardial T1 signal indicative of fat is seen in a high percentage (75%) of patients who meet the Task Force criteria for ARVD. Trabecular disarray is seen more frequently than wall thinning and aneurysms. RV dimensions and volumes differ significantly in ARVD compared to controls, indicating a role for quantitative evaluation in the diagnosis of ARVD. PMID- 12776864 TI - Arrhythmogenic right ventricular cardiomyopathy: time for a new look. PMID- 12776865 TI - Effect of amiodarone on dispersion of atrial refractoriness and cycle length in patients with atrial fibrillation. AB - INTRODUCTION: Amiodarone is effective in preventing the recurrence of atrial fibrillation (AF) after cardioversion (CV). Dispersion of atrial refractoriness may be relevant to the generation of AF. We designed a study to determine the electrophysiologic effects of amiodarone in patients with previous early recurrence of AF after CV. METHODS AND RESULTS: Fifteen patients with previous AF recurrence (without antiarrhythmic drugs) after CV (CV1) were selected for amiodarone therapy and repeat CV (CVamio). Prior to CV1, mean AF cycle length (AFCL) had been recorded at four atrial sites (right atrial appendage [RAA], distal coronary sinus [DCS], right atrial lateral wall [LAT], and interatrial septum [IAS]) and dispersion of AFCL had been calculated. These patients were treated with amiodarone and, prior to CVamio, AFCL was recorded at the four atrial sites as for CV1. Between CV1 and CVamio, AFCL increased at all atrial sites: 153 +/- 13 msec to 179 +/- 14 msec at RAA, 144 +/- 12 msec to 174 +/- 18 msec at DCS, 158 +/- 13 msec to 182 +/- 16 msec at LAT, and 161 +/- 18 msec to 181 +/- 17 msec at IAS. Dispersion of AFCL decreased from 24 +/- 10 msec at CV1 to 15 +/- 11 msec at CVamio (P = 0.01). The median time in sinus rhythm increased from 3.12 hours post CV1 to 28 days post CVamio, (P < 0.02). CONCLUSION: Amiodarone causes a reduction in the dispersion of AFCL. This action may be relevant to the beneficial effects of amiodarone in patients with AF. PMID- 12776866 TI - Clinical and electrophysiologic predictors of ventricular tachyarrhythmia recurrence in patients with implantable cardioverter defibrillators. AB - INTRODUCTION: Not all patients experience recurrent sustained ventricular tachyarrhythmias after placement of an implantable cardioverter defibrillator (ICD). We evaluated the clinical and electrophysiologic predictors of ventricular tachycardia (VT) and ventricular fibrillation (VF) recurrence following ICD implantation. METHODS AND RESULTS: Consecutive patients (n = 133) underwent 4 +/- 3 serial electrophysiologic studies (EPS) over 50 +/- 26 months following ICD implantation. Sustained VT/VF could always be induced during follow-up EPS in 49 patients; sustained VT/VF was sometimes induced during follow-up EPS in 47 patients; and sustained VT/VF could never be induced during follow-up EPS in 37 patients. Spontaneous VT/VF requiring ICD therapy occurred in 107 patients during follow-up. Patients with sustained VT/VF that was always inducible or sometimes inducible during follow-up experienced more frequent episodes of VT/VF following ICD implant (20.5, 95% CI 12.7-33.0; and 17.8, 95% CI 11.3-28.1 episodes/patient respectively; vs 3.0, 95% CI 2.0-4.6 episodes/patient for patients with VT/VF never induced, P < 0.001). Inducibility of sustained VT/VF post-ICD implant (P < 0.001) and sustained VT as the presenting arrhythmia (P = 0.02) were independent predictors of spontaneous VT/VF recurrence. CONCLUSION: Reproducibly inducible VT/VF following ICD implantation predicts a high probability of VT/VF recurrence and identifies a cohort of patients who experience frequent episodes of VT/VF over time. Persistent noninducibility of sustained VT/VF identifies a group of patients who experience no or very few episodes of VT/VF recurrence. PMID- 12776867 TI - Characterization of sustained atrial tachycardia in dogs with rapid ventricular pacing-induced heart failure. AB - INTRODUCTION: Atrial arrhythmias often complicate congestive heart failure (CHF). We characterized inducible atrial tachyarrhythmias and electrophysiologic alterations in dogs with CHF and atrial enlargement produced by rapid ventricular pacing. METHODS AND RESULTS: Endocardial pacing leads were implanted in the right ventricle, right atrium, and coronary sinus in 18 dogs. The right ventricular lead was connected to an implanted pacemaker capable of rapid ventricular pacing. The atrial leads were used to perform electrophysiologic studies in conscious animals at baseline in all dogs, during CHF induced by rapid ventricular pacing at 235 beats/min in 15 dogs, and during recovery from CHF in 6 dogs. After 20 +/- 7 days of rapid ventricular pacing, inducibility of sustained atrial tachycardia (cycle length 120 +/- 12 msec) was enhanced in dogs with CHF. Atrial tachycardia required a critical decrease in atrial burst pacing cycle length (< or = 130 msec) for induction and often could be terminated by overdrive pacing. Calcium antagonists (verapamil, flunarizine, ryanodine) terminated atrial tachycardia and suppressed inducibility. Effective refractory periods at 400- and 300-msec cycle lengths in the right atrium and coronary sinus were prolonged in dogs with CHF. Atrial cells from dogs with CHF had prolonged action potential durations and reduced resting potentials and delayed afterdepolarizations (DADs). Mitochondria from atrial tissue from dogs with CHF were enlarged and had internal cristae disorganization. CONCLUSIONS: CHF promotes inducibility of sustained atrial tachycardia. Based on the mode of tachycardia induction, responses to pacing and calcium antagonists, and presence of DADs, atrial tachycardia in this CHF model has a mechanism most consistent with DAD-induced triggered activity resulting from intracellular calcium overload. PMID- 12776868 TI - Sustained atrial tachycardia in heart failure: is it the precursor to atrial fibrillation? PMID- 12776869 TI - ZP123 increases gap junctional conductance and prevents reentrant ventricular tachycardia during myocardial ischemia in open chest dogs. AB - INTRODUCTION: The aim of this study was to determine if the stable antiarrhythmic peptide (AAP) analogue ZP123 increases gap junctional intercellular conductance and prevents reentrant ventricular tachycardia (VT) during coronary artery occlusion. METHODS AND RESULTS: Voltage clamp experiments demonstrated that 10 nM ZP123 improved gap junctional intercellular conductance by 69% +/- 20% in pairs of guinea pig ventricular myocytes. VT was induced by programmed stimulation in alpha-chloralose anaesthetized open chest dogs 1 to 4 hours after coronary artery occlusion. Three-dimensional activation mapping was done using six bipolar electrograms on each of 23 multipolar needles in the risk zone. When VT was reproducibly induced, dogs were randomly assigned to receive either saline or ZP123 cumulatively at three dose levels (intravenous bolus followed by 30-min infusion per dose). Attempts to induce VT were repeated in each infusion period. Mass spectrometry was used to measure ZP123 plasma concentrations. Twenty-six dogs with reentrant VT were included. ZP123 significantly prevented reentrant VT at all plasma concentrations vs saline: 1.0 +/- 0.2 nM: 6/12 vs 0/12; 7.7 +/- 0.6 nM: 7/13 vs 1/12; and 69.2 +/- 5.4 nM: 9/13 vs 1/13. The preventive effect of ZP123 on reentrant VT was closely correlated to reversal of functional, unidirectional conduction block. ZP123 did not affect effective refractory period, surface ECG parameters, mean arterial pressure, or infarct size. CONCLUSION: The stable AAP analogue ZP123 increased gap junctional intercellular conductance and specifically prevented the induction of reentrant VT during ischemia in a broad dose range without proarrhythmic or hemodynamic side effects. ZP123 is a promising candidate for use in preventing ischemia-induced VT. PMID- 12776870 TI - Prevention of ischemia-induced reentrant ventricular arrhythmias by a peptide that enhances gap junctional conductance. PMID- 12776871 TI - Use of electrogram characteristics during sinus rhythm to delineate the endocardial scar in a porcine model of healed myocardial infarction. AB - INTRODUCTION: Substrate-based catheter ablation of postmyocardial infarction (post-MI) ventricular tachycardia necessitates electroanatomic definition of the scarred endocardium. We sought to determine whether electrogram criteria during sinus rhythm could identify the location and extent of the myocardial scar by electroanatomic mapping. METHODS AND RESULTS: A porcine model of healed MI was generated by injecting agarose microspheres into the mid left anterior descending coronary artery. At least 4 weeks post-MI, the animals (n = 24) underwent detailed left ventricular endocardial electroanatomic mapping using a 4-mm-tip catheter (BioSense-Webster, Inc.). Based upon mapping data in normal animals, infarcted tissue was defined as bipolar electrogram amplitude < 1.5 mV and electrogram duration > or = 50 msec. Radiofrequency ablation lesions (2-10 per animal) were placed to tag the endocardial borders of the electroanatomic mapping defined scar. The area of the scar defined by abnormal voltage amplitude was 25.9 +/- 15.4 cm2 (range 6.9-60.5). This area correlated well with that defined as scar by the electrogram duration criteria (26.4 +/- 16 cm2). Of those points remote from the infarct with falsely low voltage amplitude resulting from presumed poor catheter-tissue contact, 94% were correctly identified as normal when using the electrogram duration criteria. Late potentials were observed predominantly along the borders of the infarcted myocardium. The radiofrequency lesions placed to tag the scar borders were located along the scar periphery during gross pathologic examination. CONCLUSION: During normal sinus rhythm, both bipolar electrogram voltage amplitude and electrogram duration criteria are able to help differentiate normal from scarred myocardial tissue. Using these criteria, a detailed reconstruction of the endocardial scar can be rendered by electroanatomic mapping of the heart. PMID- 12776872 TI - Substrate mapping: the historical perspective and current status. PMID- 12776873 TI - Novel concept of atrial tachyarrhythmias originating from the superior vena cava: insight from noncontact mapping. AB - INTRODUCTION: Information about the activation patterns inside the superior vena cava (SVC) and entry and exit sites at the SVC-right atrial (RA) junction during SVC tachyarrhythmia is limited. METHODS AND RESULTS: A detailed characterization of electrophysiologic mechanisms and ablation strategies was performed using a noncontact three-dimensional mapping system in two cases of SVC tachycardia. The first case demonstrated SVC tachycardia originating from an ectopic focus inside the SVC, with sustained depolarization and conduction to the atrium. Entry and exit sites across the SVC-RA junction were located very close to each other. The second case demonstrated two different reentrant circuits, one inside the SVC and the other into and out of the SVC-RA junction. The entry and exit sites were located far away from each other. CONCLUSION: Noncontact mapping may help to reveal the mechanism of SVC tachyarrhythmias and to locate entry and exit sites at the SVC-RA junction as a guide for catheter ablation. PMID- 12776874 TI - Atrial tachycardia arising from an epicardial site with venous connection between the left superior pulmonary vein and superior vena cava. AB - We describe a case of atrial tachycardia originating from an epicardial site with a venous connection between the left superior pulmonary vein (LSPV) and superior vena cava (SVC). Initial endocardial mapping with multiple electrodes catheters demonstrated early endocardial activation at both the SVC and LSPV. However, radiofrequency applications at the SVC failed to terminate the atrial tachycardia. With three-dimensional electroanatomic mapping, the earliest endocardial activation was found to be in the left atrial appendage (LAA). However, radiofrequency energy applications at multiple sites in the LAA resulted in only transient termination of the tachycardia. A left atrial angiogram demonstrated a venous connection between the LSPV and SVC, overlying the LAA. An application of radiofrequency energy with a saline-irrigated ablation catheter delivered at the earliest activation site in the LAA terminated the tachycardia. The tachycardia did not recur during 18-month follow-up. PMID- 12776875 TI - Successful catheter ablation of an anteroseptal accessory pathway from the noncoronary sinus of Valsalva. AB - We describe a patient who underwent radiofrequency catheter ablation of concealed left lateral and anteroseptal accessory pathways. After successful elimination of the concealed left anterolateral accessory pathway, the earliest retrograde atrial activation was located in the His-bundle region. Complete elimination of the accessory pathway conduction was achieved with a radiofrequency energy application from the noncoronary sinus of Valsalva. PMID- 12776876 TI - Somatic gene transfer and cardiac arrhythmias: problems and prospects. PMID- 12776877 TI - A long-RP interval tachycardia: what is the mechanism? PMID- 12776878 TI - Linear ablation in the left atrium using a nonfluoroscopic mapping system. PMID- 12776879 TI - Sinus node inhibition during ventricular stimulation. PMID- 12776880 TI - Phase singularities and termination of spiral wave reentry. PMID- 12776881 TI - Herbivore responses to plant secondary compounds: a test of phytochemical coevolution theory. AB - Literature data were collected on the floristic distribution and toxicity of phytochemicals to herbivores and on herbivore specialization in order to test phytochemical coevolution theory. The theory makes four predictions that can be tested with this information. Herbivores can adapt to novel, more toxic chemicals by becoming specialists, or they can become generalists but at the cost of lower feeding success on any particular host. Thus, the first two predictions are as follows: herbivores should do better on chemicals that are present in their normal host, and this pattern should be stronger for specialists than for generalists. The "escape and radiation" aspect of the theory holds that if a plant taxon with a novel defense chemical diversifies, the chemical will become widespread. Eventually, herbivores will adapt to and disarm it. So the third prediction is that more widespread chemicals are less toxic than more narrowly distributed ones. Because generalists should not do as well as specialists on chemicals disarmed by the latter, the fourth prediction is that the third prediction should be more true for generalists than specialists and should depend on presence/absence of the chemical in the normal host. Multiple regressions of toxicity (herbivore mortality and final weight) on three predictor variables (chemical presence/absence in the normal host, specialism, and chemical floristic distribution) and relevant interactions were used to test these predictions. Chemical presence/absence in the normal host, the interaction between this variable and specialism, and chemical floristic distribution had significant effects on both measures of toxicity, supporting the first three predictions of the model. Support for the fourth prediction (a three-way interaction among all predictor variables) was evident for final weight but not mortality, perhaps because growth is more responsive to toxicity differences than survival. In short, the phytochemistry literature provides broad support for the phytochemical coevolution model. PMID- 12776882 TI - A globally consistent richness-climate relationship for angiosperms. AB - Species richness, the simplest index of biodiversity, varies greatly over broad spatial scales. Richness-climate relationships often account for >80% of the spatial variance in richness. However, it has been suggested that richness climate relationships differ significantly among geographic regions and that there is no globally consistent relationship. This study investigated the global patterns of species and family richness of angiosperms in relation to climate. We found that models relating angiosperm richness to mean annual temperature, annual water deficit, and their interaction or models relating richness to annual potential evapotranspiration and water deficit are both globally consistent and very strong and are independent of the diverse evolutionary histories and functional assemblages of plants in different parts of the world. Thus, effects of other factors such as evolutionary history, postglacial dispersal, soil nutrients, topography, or other climatic variables either must be quite minor over broad scales (because there is little residual variation left to explain) or they must be strongly collinear with global patterns of climate. The correlations shown here must be predicted by any successful hypothesis of mechanisms controlling richness patterns. PMID- 12776883 TI - The evolution of empty flowers revisited. AB - The evolution of plants that provide no form of reward for their pollinators is puzzling because they receive low numbers of pollinator visits and so have low reproductive success. To predict the evolutionary dynamics of empty morphs within a plant population, we modeled different foraging strategies that pollinators could use to avoid them. We predicted that the optimal strategy was to visit empty inflorescences randomly when these were infrequent but to use strategies such as visiting fewer flowers per inflorescence to avoid wasting time on them. As the frequencies of empty inflorescences increased, discriminating directly against empty morphs was more likely to be an optimal strategy than was avoiding the species altogether and switching to an alternative one. An experimental test of this model using artificial inflorescences showed that bumblebees used a variety of strategies to minimize time wasted on empty inflorescences. They showed weak discrimination against empty inflorescences but switched to an alternative type of inflorescence as the frequency of empty inflorescences increased. We predicted that empty morphs would be at a visitation rate disadvantage even when at low frequencies in a plant population. Differences in outcrossing rates, or male function, may explain how rewardlessness spreads in a plant population. PMID- 12776884 TI - On the heritability of geographic range sizes. AB - Within taxonomic groups, most species are restricted in their geographic range sizes, with only a few being widespread. The possibility that species-level selection on range sizes contributes to the characteristic form of such species range size distributions has previously been raised. This would require that closely related species have similar range sizes, an indication of "heritability" of range sizes at the species level. Support for this view came from a positive correlation between the range sizes of closely related pairs of fossil mollusc species. We extend this analysis by considering the relationship between the geographic range sizes of 103 pairs of contemporary avian sister species. Range sizes in these sister species show no evidence of being more similar to each other than expected by chance. A reassessment of the mollusc data also suggests that the high correlation was probably overestimated because of the skewed nature of range size data. The fact that sister species tend to have similar life histories and ecologies suggests that any relationship between range sizes and biology is likely to be complicated and will be influenced by historical factors, such as mode of speciation and postspeciation range size transformations. PMID- 12776885 TI - How localized consumption stabilizes predator-prey systems with finite frequency of mixing. AB - Predator-prey theory began with aspatial models that assumed organisms interacted as if they were "well-mixed" particles that obey the laws of mass action, but it has become clear that both the spatial and individual nature of many organisms can change how the dynamics of such systems function. Here I examine how localized consumption of prey by predators changes the dynamics of predator-prey systems; I use an individual-based simulation of the Rosenzweig-MacArthur model in implicit space and its mean-field approximation. In combination with limited movement, localized consumption makes the predator-prey dynamics more stable than the comparable "well-mixed" Rosenzweig-MacArthur model. Using a spatial correlation, one can directly compare a simplified version of the individual based model with the Rosenzweig-MacArthur model. While this comparison allows the changes in the dynamics to be captured by the "well-mixed" Rosenzweig-MacArthur model, the parameters of the functional response are now dependent on the movement parameters, and so the functional response must be estimated statistically from the dynamics of the individual-based model. Yet this implies that aspatial models may work in a scale-specific fashion for spatial systems. Unlike many recent spatial models, the localized consumption and limited movement in the model presented here cannot produce coherent spatial patterns and do not depend on a patchy structure, as found in metapopulation models. Instead, the individual nature of the interactions creates a diffusion-limited reaction, which appears closer to a form of ephemeral refuge. PMID- 12776886 TI - Integrating the roles of information and competitive ability on the spatial distribution of social foragers. AB - Understanding and predicting the spatial distribution of social foragers among patchily distributed resources is a problem that has been addressed with numerous approaches over the 30 yr since the ideal free distribution (IFD) was first introduced. The two main approaches involve perceptual constraints and unequal competitors. Here we present a model of social foragers choosing among resource patches. Each forager makes a probabilistic choice on the basis of the information acquired through past foraging experiences. Food acquisition is determined by the forager's competitive ability. This model predicts that perceptual constraints have a greater influence on the spatial distribution of foragers than unequal competitive abilities but that competitive ability plays an important role in determining an individual's information state and behavior. Better competitors have access to more information; consequently, we find that competitive abilities and perceptual constraints are integrated through the social environment occupied by individual foragers. Relative competitive abilities influence the forager's information state, and the ability to use information determines the resulting spatial distribution. PMID- 12776887 TI - Biological correlates of extinction risk in bats. AB - We investigated patterns and processes of extinction and threat in bats using a multivariate phylogenetic comparative approach. Of nearly 1,000 species worldwide, 239 are considered threatened by the International Union for Conservation of Nature and Natural Resources (IUCN) and 12 are extinct. Small geographic ranges and low wing aspect ratios are independently found to predict extinction risk in bats, which explains 48% of the total variance in IUCN assessments of threat. The pattern and correlates of extinction risk in the two bat suborders are significantly different. A higher proportion (4%) of megachiropteran species have gone extinct in the last 500 years than microchiropteran bats (0.3%), and a higher proportion is currently at risk of extinction (Megachiroptera: 34%; Microchiroptera: 22%). While correlates of microchiropteran extinction risk are the same as in the order as a whole, megachiropteran extinction is correlated more with reproductive rate and less with wing morphology. Bat extinction risk is not randomly distributed phylogenetically: closely related species have more similar levels of threat than would be expected if extinction risk were random. Given the unbalanced nature of the evolutionary diversification of bats, it is probable that the amount of phylogenetic diversity lost if currently threatened taxa disappear may be greater than in other clades with numerically more threatened species. PMID- 12776888 TI - Parental effects in a partially self-incompatible herb Campanula rapunculoides L. (Campanulaceae): influence of variation in the strength of self-incompatibility on seed set and progeny performance. AB - We employ a full reciprocal diallel design between 10 parental plants that differed in their strength of self-incompatibility (SI; strong, intermediate, and weak) to examine parental effects on seed set and 10 components of fitness of progeny performance in Campanula rapunculoides. We perform ANOVAs to separate the influence of the strength of SI and the identity of the maternal and paternal parent on family performance. We calculate the phenotypic and genetic correlations between traits to determine potentially evolutionary constraints. Finally, we employ maximum likelihood methods to estimate the components of quantitative genetic variance, as defined by Cockerham and Weir in their BioModel c. Our most significant finding is that weak SI plants have high outcrossed seed set as maternal parents. We argue that direct or pleiotropic effects of modifiers of SI probably cause this. Second, we find that extranuclear interactions, as defined by the BioModel, have strong effects on seed set and several vegetative and flowering traits. These findings indicate that some maternal plants selectively provision seeds sired by specific paternal donors and that some of this variation appears to be associated with modifiers of the strength of SI. We find other sources of significant quantitative genetic variation for all of the traits we examine and discuss the possible role these play in the evolution of the reproductive system. Taken together, our findings show that variation in the strength of SI may influence levels of quantitative genetic variation that, in turn, can influence the reproductive success of individuals in C. rapunculoides PMID- 12776889 TI - Local extinction and the evolution of dispersal rates: causes and correlations. AB - We present the results of individual-based simulation experiments on the evolution of dispersal rates of organisms living in metapopulations. We find conflicting results regarding the relationship between local extinction rate and evolutionarily stable (ES) dispersal rate depending on which principal mechanism causes extinction: if extinction is caused by environmental catastrophes eradicating local populations, we observe a positive correlation between extinction and ES dispersal rate; if extinction is a consequence of stochastic local dynamics and environmental fluctuations, the correlation becomes ambiguous; and in cases where extinction is caused by dispersal mortality, a negative correlation between local extinction rate and ES dispersal rate emerges. We conclude that extinction rate, which both affects and is affected by dispersal rates, is not an ideal predictor for optimal dispersal rates. PMID- 12776890 TI - The evolution of androgenesis. AB - It is well known that some species produce offspring carrying only female chromosomes by processes such as apomixis and parthenogenesis (generically termed "gynogenesis"). There are also several cases of natural reproduction by androgenesis in which diploid offspring carry nuclear chromosomes from only the male parent. We used population genetics models to investigate the conditions for invasion of rare androgenesis alleles and the consequences of their spread. Our models predict that androgenesis alleles often spread to fixation. If fixation causes the loss of females or female function in the population, population extinction occurs. Therefore, androgenesis alleles represent a new class of selfish genetic elements. Extinction is more likely in dioecious species than in hermaphrodites. Within dioecious species, extinction is more likely when androgenesis occurs via paternal apomixis (vs. fusion or doubling of haploid nuclei) and when females are the heterogametic sex (vs. male heterogamety). The apparent rarity of androgenesis compared to gynogenesis could be because androgenesis is harder to detect and more often leads to population extinction. Also, there could be greater evolutionary constraints on the origin of mutations for androgenesis. We suggest characteristics of groups in which further cases of androgenesis are more likely to be found. PMID- 12776891 TI - Changes in vigilance with group size under scramble competition. PMID- 12776892 TI - Pattern does not equal process: exactly when is sex environmentally determined? PMID- 12776893 TI - Iron deficiency anemia: preventive strategies and controversies. PMID- 12776894 TI - Preventive strategy to control iron deficiency anemia in children and adults. AB - OBJECTIVE: To compare improvement in hemoglobin, mean corpuscular volume, mean corpuscular hemoglobin and ferritin levels in children aged 5-10 years and women of reproductive age (15-45) supplemented with daily oral and once weekly with single and double dose of ferrous sulphate. METHOD: Twenty children received 200 mg ferrous sulfate daily and 20 received the same dose once weekly for two months. Ten women received 300 mg ferrous sulfate daily, 10 received the same dose once weekly while 10 received 600 mg of ferrous sulfate once weekly for one month. RESULTS: All parameters improved significantly in children who received 200 mg ferrous sulfate daily and weekly. Similarly, the parameters improved significantly in women who received 300 mg of ferrous sulfate daily and 600 mg of ferrous sulfate weekly. CONCLUSION: Weekly supplementation of iron is far better in controlling iron deficiency anemia due to cost effectiveness and better compliance. PMID- 12776895 TI - Association of iron deficiency anaemia in children of 1-2 years of age with low birth weight, recurrent diarrhoea or recurrent respiratory tract infection--a myth or fact? AB - AIMS AND OBJECTIVE: To investigate for the presence or absence of any association between low birth weight, recurrent diarrhoea or recurrent acute respiratory infections with iron deficiency anaemia in Pakistani children aged 1-2 years. METHODOLOGY: From July 1993 to July 1995 a case control study was conducted at the Community Health Centre (CHC). Fifty cases and 100 controls were included. Informed consent was taken from mothers for the blood tests of their children and a questionnaire was administered to them. Data obtained from the completed questionnaires was analyzed by chi-square and t-tests. RESULTS: There was no statistically significant difference in low birth weight (p = 0.712), recurrent diarrhoea or respiratory infections between anaemic and non-anaemic children at 1 2 years age (OR of 1.71 and 1.48 respectively). CONCLUSION: This study does not uphold the belief that low birth weight, recurrent diarrhoea or respiratory infections have a causal association with iron deficiency anaemia in 1-2 years children. PMID- 12776896 TI - Hepatitis B and C single and co-infection in chronic liver disease and their effect on the disease pattern. AB - OBJECTIVE: To find the frequency of hepatitis B and C virus (HBV and HCV) infection and their coinfection in chronic liver disease and any differences in the clinical features of single and coinfections. METHODOLGY: All clinically recognisable adult CLD patients admitted to Military Hospital, Rawalpindi in years 1999-2000, were included in the study. Their history and clinical features were recorded. Seromarkers for HBV (HBsAg, anti-HBc) and HCV (anti-HCV) were done. RESULTS: A total of 97 (52 male, 45 female) patients were included in the study. Mean age was 51.6 years (range 16-75 years). Cirrhosis was present in 74.2%, chronic hepatitis (Ch Hep) in 18.6% and hepatocellular carcinoma (HCC) in 7.2%. Among the 74.1% patients with cirrhosis 28.5% were HBsAg positive, 55.5% anti-HBc positive and 68.1% anti HCV positive. In the chronic hepatitis cases 27.8% were HBsAg positive, 23.6% anti-HBc positive and 61.1% anti HCV positive. Among the patients of HCC 59.7% were HBsAg positive, 71.4% anti-HBc positive and 42% anti HCV positive. Regarding coinfection 7.2% were anti-HCV and HBsAg positive while 35.1% were anti-HCV and anti-HBc positive. CONCLUSION: Ninety percent of patients with CLD had evidence of HBV, HCV or coinfection. Disease was more severe in patients with coinfection. Anti-HBc was found to be a sensitive indicator of past HBV. PMID- 12776897 TI - Chronic renal failure in children. AB - OBJECTIVE: To determine the clinical presentation and find out the underlying cause of chronic renal failure (CRF). METHODS: This is a descriptive prospective study of patients attending children hospital, Chandka Medical College Larkana and National Institute of Child Health (Paediatric Nephrology Unit) Karachi from January 1993 to December 1996. RESULTS: Seventy eight confirmed cases of chronic renal failure (CRF) were included in the study. Out of them 51 were male and 27 females (M:F ratio 1.8:1). Most of the children (90%) presented after 3 years of age. Common clinical presentation were anaemia (93.5%) cases, growth retardation (74.3% cases), urinary complaints (65.3%) cases and hypertension (53.8%) cases. Common causes leading to CRF were reflux nephropathy (24.5%), posterior uretheral valves (23%), chronic glomerulonephritis (15.4%) and urolithiasis (14%). CONCLUSION: Early diagnosis and management of most common causes of CRF may prevent and delay the progression to end stage renal disease (ESRD). PMID- 12776898 TI - Translation and cultural adaptation of health questionnaires. AB - BACKGROUND: Health research in Pakistan often requires questionnaires in English language developed in the West to be translated into the local language. Many of the factors measured by these questionnaires are complex and apply to a different culture. Simple translations may lead to problems of validity and reliability in the Pakistani setting. This paper describes the strategies adopted for the translation and cross-cultural adaptation of the Self-Reporting Questionnaire (SRQ), a screening questionnaire for mental health developed by the World Health Organisation. METHODS: A general protocol was developed for translation of questionnaires into Urdu, describing each step in the translation procedure. Key informant interviews were carried out to obtain better cultural understanding of difficult concepts. The translation was tested and disputed items discussed in a focus group in a structured manner. RESULTS: Modifications were made to the questionnaire in light of the target population's culture and language. CONCLUSION: Simple translations are often insufficient for complex questionnaires. Key informant interviews and focus groups are useful to address conceptual and construct issues in such questionnaires. PMID- 12776899 TI - Radiosurgery for the control of glomus jugulare tumours. AB - OBJECTIVE: To ascertain the efficacy of stereotactic radiosurgery (gamma knife) for the control of glomus jugulare tumours. METHOD: Between March 1994 and December 1997 we treated eight patients of glomus jugulare tumour with radiosurgery. These patients have been followed for more than four years (range 52 to 97 months). The age of the patients ranged between 32-64 years (mean 53 years). The male: female ratio was 3:5. Three patients had previously undergone surgery and one had unsuccessful embolization. The dose applied to tumour margin ranged between 16-25 Gy (median 25 Gy). Patients were followed up with yearly MRI scans and where possible with cerebral angiography. RESULTS: All patients showed stabilisation of their symptoms following radiosurgery and six improved clinically. Five of these patients showed decrease in the size of lesion seen objectively on radiology, either slight to moderate decrease seen on the MRI scan or reduction in size and vascularity seen on cerebral angiography. The procedure is minimally invasive and none of the patients showed any adverse effect to radiosurgery. CONCLUSION: These results are encouraging but because of its naturally slow growth rate, up to 10 years of follow up will be necessary to establish a cure rate after radiosurgery for these lesions. PMID- 12776900 TI - Critical appraisal of endometriosis management for pain and subfertility. AB - OBJECTIVE: To critically appraise current best available management of pain and subfertility ascribable to endometriosis. METHODS: Medline and specialist computer databases were searched along with reference lists of known reviews and primary articles to identify cited articles not captured by electronic searches. We included all studies pertaining to treatment of pain and subfertility ascribable to endometriosis. DISCUSSION: Medical and surgical management options for pain and subfertility are presented as Evidence-based inquiries for critically appraised topics. Each problem is presented as an answerable question followed by assessment of literature search for level of evidence. This is followed by critical appraisal of results. In the end, take-home answer to individual problem is presented in view of best available evidence. CONCLUSION: Evidence-based management of endometriosis provides realistic therapeutic goals and expectations for the clinicians and women experiencing pain and subfertility due to this disease. It also promotes efficient and effective use of medical and surgical options when required. PMID- 12776901 TI - Clinical decision making Part I: errors of commission and omission. AB - Clinical decision making refers to any act of diagnosis that leads to a decision regarding prognosis, treatment, referral or counseling. It has long been recognized that diagnostic probabilities influence perceptions and judgements in medical practice. However, so do ethical values affect clinical judgements, even if these lead to adverse consequences: for example, to judge a well person sick is considered "more ethical" than to judge a sick person well. This dilemma confronts us also when having to choose between what are called Type 1 and Type 2 errors, and the consequences of either may be serious. Other factors that should be taken into account, as they can affect the "pretest likelihood" or "prior probability" of disease and the error rates obtained, include the individual circumstances of the patient (e.g. socioeconomic background), the organizational setting (e.g. patient volume, cultural practices and norms), and system differences (e.g. fee for service versus salaried service). In Part II of this two-part series,we explore in greater depth the mathematical basis for why various diagnostic procedures perform differently in different settings. PMID- 12776903 TI - Do we need daily iron supplementation? Comments and controversies. PMID- 12776902 TI - Moyamoya disease: an elusive diagnosis. AB - OBJECTIVE AND BACKGROUND: Moyamoya disease is an idiopathic vasculopathy of circle of Willis. Stroke is a common presentation. We describe clinical and radiological features of moyamoya disease in four patients. SETTING: Tertiary care center in a metropolitan city. MATERIALS AND METHODS: Patients with moyamoya disease were identified by ICD-9 coding system of the hospital medical records. Demographic characteristics, clinical features/presentation, laboratory investigations and radiological investigations were recorded and analyzed. RESULTS: Four patients (three children and one adult) presented with hemiparesis. One patient expired during hospital stay. Two patients underwent surgery (synangiosis). EEGs of three patients showed focal abnormalities. Neuroimaging showed ischemic infarctions in all patients. Findings on four vessel digital substraction angiography and/or magnetic resonance angiography were compatible with moyamoya disease. CONCLUSION: Moyamoya disease should be considered in all young patients, especially children, presenting with stroke. PMID- 12776904 TI - Persistent hyperplastic primary vitreous. PMID- 12776906 TI - When antibodies are antigens. PMID- 12776905 TI - An evaluation of topical and local anesthesia in phacoemulsification. AB - OBJECTIVE: To assess the comparative efficacy of topical and local anesthesia in phacoemulsification. MATERIAL AND METHODS: The medical records of 186 men and women between the ages of 45-85 years, who underwent elective cataract surgery by phacoemulsification technique, under the care of one surgeon, over a period of one year, from March 1999-March 2000 were reviewed. RESULTS: Of 186 patients who underwent small incision, self-healing phacoemulsification cataract surgery, 124 received topical and 62 local anesthesia. The most common cataract types were nuclear sclerosis. The duration of surgery with topical anesthesia was shorter. Sutures and eye padding were more frequently applied for procedures done under local anesthesia. Uncorrected visual acuity in the first post-op week was between 20/20-20/50 for 53.6% of the cases done under topical compared to 30.9% in local anesthesia. A similar trend was noted in the visual acuity one month post operatively. CONCLUSION: The uncorrected visual acuity improves faster and the duration of surgery is shorter when topical anesthesia is used. PMID- 12776907 TI - How endothelial cell organo-specificity mediates circulating cell homing. AB - Normal and transformed cells home into tissues from the circulation in a very selective way thanks to highly complex molecular mechanisms that govern cell-to cell interactions and drive the homing of circulating cells so that it is achieved properly. Because this is characterized by a resulting high selectivity, it constitutes a template for targeted drug-, gene- or cell-therapy strategies. Designing a mimetic-based therapy requires the identification of the responsible selective molecules, but also their mechanisms of action and interactions with their ligands together with their biological modulation and regulation. This homing/invasion event is decisive at the level of the endothelium that lines the vessel walls. Since cell-to-cell interactions mean a double recognition process, this review will illustrate the part played by the endothelial cells (ECs) and their adhesion molecules: the protein as well as the glycan point of view, the chronology, and the environmental modulation of EC adhesion molecule expression. These characteristics should provide keys to understanding the resulting overall specificity of cell localization. Taking into account the cytokine microenvironment, a fundamental role was recently documented for locally secreted chemokines which act through their restricted presentation by endothelial cells. As such, chemokines contribute to illustrating the concept of endothelial organo specificity which is approached here, uncovering the role of glycoconjugate signaling as the hallmark of refined cellular recognition, and discussed in the context of potential drug design against site-directed diseases such as metastases, inflammatory leukocyte recruitment, and tumor/inflammation-induced angiogenesis. PMID- 12776908 TI - Suppressors of cytokine signaling proteins in innate and adaptive immune responses. AB - Suppressors of cytokine signaling (SOCS) proteins have been identified as important mediators of negative regulatory circuits within cytokine receptor signaling. They are induced upon stimulation by an increasing set of cytokines as well as further immunological stimuli and are capable to inhibit Janus kinases and signal transducer and activator of transcription signaling. Inhibition is mediated by interfering directly with signal transduction at the receptor as well as targeting of associated molecules for proteosomal degradation. Targeted gene deletion approaches have revealed the importance of SOCS mediated termination of cytokine signaling during normal cellular activation. In addition to their function as classical feedback inhibitors SOCS proteins display a broad panel of inhibitory activity thereby mediating cross-talk modulation between different stimuli. The consequences for regulation of innate and adaptive immune responses are thus obvious. Finally, there are emerging data showing involvement of SOCS proteins in various immune diseases. Modulating SOCS activity could be a promising new approach for molecular therapeutic strategies. PMID- 12776909 TI - From inflammation to sickness: historical perspective. AB - The concept of the four cardinal signs of acute inflammation comes from antiquity as rubor et tumor cum calore et dolore, (redness and swelling with heat and pain) extended later by functio laesa (loss of function). The contemporary understanding of this process we owe to 19th-century milestone discoveries by Rudolph Virchow, Julius Cohnheim and Elie Metchnikoff. In the 20th century, the development of potent technological tools allowed the rapid expansion of knowledge of the cells and mediators of inflammatory processes, as well as the molecular mechanisms of their interactions. It turned out that some mediators of inflammation have both local and distant targets, among them the liver (responding by the production of several acute phase reactants) and neurohormonal centers. In the last decades it has become clear that the immune system shares mediators and their receptors with the neurohormonal system of the body; thus, they form a common homeostatic entity. Such an integrative view, introduced by J. Edwin Blalock, when combined with Hans Selye's concept of stress, led to the contemporary understanding of sickness behavior, defined by Robert Dantzer as a highly organized strategy of the organism to fight infections and to respond to other environmental stressors. PMID- 12776910 TI - The dynamic and complex role of mast cells in allergic disease. AB - Mast cells (MCs) are found widely distributed in tissues and contribute to regulation of inflammatory responses and ongoing modulation of the tissues. Although MCs are important in a variety of processes, including innate immunity, their role in allergic disease has received increasing attention in the past decade. MCs are located throughout the human body and, upon allergen exposure, they are stimulated via the immunoglobulin E (IgE) receptor (Fc(epsilon)RI) to release several pro-inflammatory mediators such as tumor necrosis factor (TNF), reactive oxygen species such as nitric oxide (NO), proteases, and lipid-derived mediators. However, we now recognize that MCs can be activated by a variety of mechanisms and that mediator release is a consequence of several intra- and extracellular signals. Some of these mechanisms, such as Fc receptor aggregation and proteinase-activated receptor (PAR)-mediated activation facilitate and augment local inflammatory responses. Other mechanisms, such as interferon gamma (IFN-gamma) induction of NO, may inhibit MC function and downregulate inflammatory responses. Increased understanding of these complex pathways has encouraged the development of therapies for allergic inflammation that target specific MC functions and mediators. Some novel strategies include oligonucleotides that induce or inhibit the production of specific mediators. Such approaches may yield useful therapies for allergic individuals in the near future. PMID- 12776911 TI - NK T cell-NK cell cross-talk: reciprocal interaction and activation? AB - Initiation and propagation of the immune response is the result of a series of coordinated cellular and biochemical interactions that lead to the activation of multiple cell types. It is now clear that an optimal immune response requires precise and rapid communication between different cell subsets. This phenomenon, referred to as cross-talk, is believed to be an essential component of the immune response that provides necessary inflammatory mediators and cytolytic activity for controlling infections and diseases. An example of effective cooperation between different cell types has been recently illustrated by the finding that specific activation of CD1-restricted natural killer T cells (NK T) can quickly lead to the activation of other subsets of cells, such as natural killer (NK) and CD8 T cells. PMID- 12776912 TI - Distribution of HLA-C alleles determined by PCR-SSP in the population of Lower Silesia. AB - We typed 100 unrelated, healthy Poles from the region of Lower Silesia for HLA-C using low-resolution polymerase chain reaction sequence-specific primers (PCR SSP) and compared the observed allele frequencies with data published for other human populations. Poles appeared to be most similar to the Germans and English more distant from the French, Catalans and Basques and most dissimilar to non Caucasians from Equatorial Guinea and Japan. It would be interesting to HLA-C type other Slavic and non-Slavic peoples from Middle and Eastern Europe for comparison. PMID- 12776914 TI - Study of thiol consumption by yeast lees. AB - The capacity of yeast lees to remove thiols was investigated in a synthetic medium and in wine. In a synthetic medium, yeast lees are able to reduce thiol concentration with a concomitant production of a disulfide. In wine, thiol consumption is lower than in the synthetic medium and no disulfide formation occurs. Though free SH units seem to be partially involved in the thiol consumption process, this does not seem to require molecular oxygen. Since thiol consumption and disulfide formation have been shown to be inhibited by EDTA, it is suggested that metallic cations may be involved in both phenomena. PMID- 12776913 TI - Serological differentiation of Helicobacter pylori CagA(+) and CagA(-) infections. AB - Many Helicobacterpylori strains causing gastroduodenal diseases have a cagA gene encoding CagA protein, a virulence factor of these bacteria. Anti-CagA antibodies produced by the majority of people infected with CagA(+) strains can indicate such an infection. In this study, the efficacy of three immunoenzymatic tests for detecting CagA(+) and CagA(-) infections were compared: immunoblot (Milenia ID Blot H. pylori IgG; MB) and ELISA conducted either with a recombinant immunodominant fragment of CagA (rCagA) or the full-length CagA molecule (flCagA). The 13C-urea breath test (13C-UBT) was used for establishing H. pylori status. The serum samples from 157 individuals were used for serodiagnosis. H. pylori CagA(+) infection was detected in H. pylori-infected individuals with similar frequencies by MB (64%) and flCagA-ELISA (60%) and a little less frequently by rCagA-ELISA (53%). There was a high coincidence between the negative results of these three tests for H. pylori-uninfected individuals with no anti-CagA IgG in the serum (96-100%). The results show that rCagA-ELISA and, especially, flCagA-ELISA are easy, inexpensive and useful noninvasive assays for the discrimination of CagA(+) and CagA(-) H. pylori infections in subjects examined by urea breath test. PMID- 12776915 TI - Growth profile of ectomycorrhizal fungal mycelium: emphasis on substrate pH influence. AB - A knowledge of behaviour of ectomycorrhizal fungal isolates to substrate pH would help in identifying candidate fungi for plantation programs. Ectomycorrhizal fungus isolates were studied to determine the pH optima for growth, substrate acidification by the culture and the effect of substrate acidification on culture growth. Of the isolates tested, the members of Agaricales (except Laccaria laccata) and Aphyllophorales favored neutral to near neutral pH, while members of the order Sclerodermatales strictly favored acidic pH. The change in substrate pH (deltapH) was maximum at the optimum growth pH for acidophilic isolates but minimum for neutro/basophilic isolates. The experiments indicate that the substrate pH would not only determine the growth rate of the fungus but also limits further proliferation of the fungus in medium. This gives vital information for determining the subculture frequency and for designing substrate parameters for nursery/plantation programs. PMID- 12776916 TI - Chromosomal polymorphism in the yeast species Debaryomyces hansenii. AB - Pulse field gel electrophoresis karyotypes of 41 strains of the genus Debaryomyces, including 35 strains confirmed as D. hansenii species by D1/D2 ribosomal DNA sequence analysis, were performed. Electrophoretic karyotypes of the 41 strains exhibited 4 to 10 chromosomal bands ranging between 0.7 Mb and 4.2 Mb. Among D. hansenii species, the patterns of strains obtained from the CBS collection and cheese isolates differed strongly from D. hansenii var. hansenii CBS767T. Both D. hansenii var. hansenii and D. hansenii var. fabryii showed chromosome length polymorphism. Electrophoretic karyotypes of the D. hansenii strains were analyzed by Southern hybridization with various species-specific probes isolated from D. hansenii var. hansenii CBS767T. Repeated sequences including the F01pro, M18pro, the Ty1-copia retrotransposon Tdh5 and hypothetical telomeric sequence hybridized to several chromosomal bands, while a D1/D2 probe derived from the large ribosomal sub-unit hybridized only to the largest chromosome. Unique probes such as those hybridizing to actin ACT1, glycerol-3 phosphate dehydrogenase GPD1 and beta-glucosidase LAC4 encoding genes were assigned to specific chromosomal bands of D. hansenii var. hansenii CBS767T. These probes failed to hybridize to D. hansenii var. fabryii strongly suggesting that strains of this variety actually represent a different taxon. PMID- 12776918 TI - Cryptococcus antarcticus var. circumpolaris var. nov., a basidiomycetous yeast from Antarctica. AB - Cryptococcus antarcticus Vishniac & Kurtzman var. circumpolaris Vishniac and Onofri var. nov. (Filobasidiales, Tremellomycetidae, Hymenomycetes), an anamorphic yeast with ca. 73% nDNA relatedness to Cryptococcus antarcticus var. antarcticus, differs in failure to assimilate raffinose, a lower maximum temperature for growth, fatty acid profile, and in a single nucleotide change in the D2 region of LSU rDNA. PMID- 12776917 TI - New preparation of PM2 phage DNA and an endonuclease assay for a single-strand break. AB - PM2 is a bacteriophage which has closed circular double-stranded DNA as a genome, which is the sole source for endonuclease assay for a single strand break in the fmol range. Therefore, it is important to isolate PM2 DNA with low control nicks for the endonuclease assay. Usually, the isolation method of phage DNA is to use ultracentrifugation which takes at least 4 days. In this report, a fast and effective method which takes only 2 days was developed to purify DNA using polyethylene glycol (PEG) 8000 and the yields of phage DNA isolated by these two methods were compared. The method using PEG 8000 increased the yield of PM2 DNA from 31.2% to 45.2%, and decreased the nick from 17.1% to 13.1%. Recently, the complete PM2 DNA genome sequence of 10,079 bp was published. The exact number of nucleotides of PM2 DNA is important for the correct enzyme assay which measures nicks generated by an endonuclease. The correct calculation of endonuclease activity of rpS3 for nick-circle assay was performed to measure single-strand breaks in this report. PMID- 12776919 TI - Defective zoospore encystment and suppressed cyst germination of Phytophthora palmivora caused by transient leaching treatments. AB - The behaviour of encysting zoospores of Phytophthora palmivora during leaching conditions was studied. Zoospores encysted and germinated successfully on polycarbonate membranes after mechanical agitation. Transient (10 min) leaching treatments with nutrient-free buffer underneath the membranes resulted in abnormal encystment and poor germination. The disruption was greatest when leaching was applied during the first minutes after start of encystment and not observed after 20 min. The early sensitivity of cells to leaching coincided with the period when alkali-resistant cell walls were formed (2-6 min after mechanical agitation). Effects of calcium and organic nutrients on encystment during leaching and germination after these treatments were studied. The disruption of encystment by early leaching treatments, but not the suppression of cyst germination, was overcome by adding calcium chloride during mechanical agitation of zoospores. Leaching with calcium containing buffer resulted in suppressed cyst germination as was the case with buffer alone. Leaching with 0.1% peptone containing buffer promoted consistently high encystment and germination. PMID- 12776920 TI - Classification of novel soil streptomycetes as Streptomyces aureus sp. nov., Streptomyces laceyi sp. nov. and Streptomyces sanglieri sp. nov. AB - The taxonomic positions of soil isolates known as Streptomyces groups A, B and C were clarified. Comparative 16S rDNA sequence studies indicated that representatives of all three taxa formed distinct phyletic lines within the Streptomyces tree though the group A strains were shown to be related to Streptomyces griseus and associated validly described species. The taxonomic integrity of all three groups was highlighted by DNA:DNA relatedness and ribotype data though the group A strains encompassed a higher degree of genetic variation than the group B and C strains. In light of these and earlier phenotypic data it is proposed that Streptomyces groups A, B and C be given species status as Streptomyces sanglieri sp. nov., Streptomyces aureus sp. nov. and Streptomyces laceyi sp. nov., respectively. PMID- 12776921 TI - Bullera anomala sp. nov. and Bullera pseudovariabilis sp. nov., two new ballistoconidium-forming yeast species from Yunnan, China. AB - Two ballistoconidium-forming yeast strains, CH 2.504 and CH 2.509, were isolated from the wilting leaves of Parthenocissus sp. collected in Yunnan, China in 1996. These two strains were assigned to the genus Bullera Derx by the characteristics of forming rotationally symmetric ballistoconidia, containing xylose in the cell hydrolysates and having Q-10 as the major ubiquinone. Molecular phylogenetic analysis based on small subunit ribosomal RNA gene (18S rDNA) and internal transcribed spacer (ITS) region sequences revealed that CH 2.504 is closely related to B. variabilis Nakase & Suzuki, while CH 2.509 is located in a clearly separate clade among the described Bullera species and related taxa. The results indicated that CH 2.509 represents a new species, for which Bullera anomala sp. nov. is the proposed name (type strain AS 2.2094T). Further DNA-DNA reassociation data showed that CH 2.504 also represents a distinct species, for which the name Bullera pseudovariabilis sp. nov. is given (type strain AS 2.2092T). PMID- 12776922 TI - Evidence for phylogenetic inheritance in pathogenicity of Mycobacterium. AB - In this study, we attempt to highlight part of the adaptive and phylogenetic constraints in mycobacterial pathogenicity. For this purpose, we first provide a phylogeny of Mycobacteria based on cladistic analyses of 64 different taxa. We then performed a comparative analysis, taking into account both ecological factors and phylogenetic relationships. The GLIM modelling analysis showed that different ecological and phylogenetic factors might be invoked to explain the variation in pathogenicity levels. Interestingly, the most harmful species were shown to be connected with the most diversified habitats. However, the independent contrast analysis revealed that once phylogeny was taken into account, none of the relationships between ecological factors and pathogenicity remained significant, and the pathogenicity appeared to be phylogenetically inherited among mycobacteria. The most pathogen were found in the slow growing/long helix 18 group, and within this group in the most derived taxa. PMID- 12776923 TI - Combined solvent and water activity stresses on turgor regulation and membrane adaptation in Oceanimonas baumannii ATCC 700832. AB - Oceanimonas baumannii ATCC 700832 is a Gram negative marine bacterium capable of utilising phenol as asole carbon source. The ability of the bacterium to tolerate low water activity when utilising either succinate or phenol as a substrate in minimal medium was studied. The membrane lipid and protein composition showed two discreet adaptive phases as salinity increased. Firstly, when NaCl concentration was increased from 0.15% (w/v), the minimum at which growth was observed, to 1% NaCl (w/v), the ratio of zwitterionic to anionic phospholipids in the membrane increased significantly. At the same time the ratio of saturated to unsaturated fatty acids and the total membrane protein decreased significantly. The second phase was observed when salinity was increased from 1% to 7% NaCl (w/v) as the ratio of zwitterionic to anionic phospholipids decreased and membrane protein increased. However, the ratio of saturated to unsaturated fatty acids was unaffected. Salinity also affected the tolerance of cultures to elevated levels of phenol. Cultures grown in 0.15% NaCl (w/v) could tolerate 12 mM phenol, whereas in the presence of 1% NaCl (w/v) cultures continued to grow in up to 20 mM phenol and in 7% NaCl (w/v) cultures 8 mM phenol could be tolerated. Changes to the composition of the membrane phospholipids and fatty acids were also observed when phenol concentrations were at the maximum that could be tolerated. Under such conditions the ratio of zwitterionic to anionic phospholipids decreased twofold compared to cultures utilising 4 mM phenol as the substrate, in all salinities except in 7% NaCl (w/v) cultures, where there was no significant effect. The ratio of saturated to unsaturated fatty acids increased significantly in all salinities compared to cultures grown with 4 mM phenol. PMID- 12776924 TI - Prevalence of 1-aminocyclopropane-1-carboxylate deaminase in Rhizobium spp. AB - This is the first report documenting the presence of 1-aminocyclopropane-1 carboxylate (ACC) deaminase in Rhizobium. This enzyme, previously found in free living bacteria, yeast and fungi, degrades ACC, the immediate precursor of ethylene in higher plants. Thirteen different rhizobial strains were examined by Southern hybridization, Western blots and ACC deaminase enzyme assay. Five of them tested positive for ACC deaminase. Induction of the expression of ACC deaminase was examined in one of the positively tested strains, Rhizobium leguminosarum bv. viciae 128C53K. This rhizobial ACC deaminase had a trace basal level of expression without ACC, but could be induced by a concentration of ACC as low as 1 microM. The more ACC added to this Rhizobium the higher the expression level of the ACC deaminase. PMID- 12776925 TI - A new hepatoma cell line for toxicity testing at repeated doses. AB - Many cell models that are used to assess basic cytotoxicity show a good correlation with acute toxicity. However, their correlation with the toxicity seen following chronic in vivo exposure is less evident. The new human hepatoma cell line HBG BC2 possesses the capacity of being reversibly differentiated in vitro and of maintaining a relatively higher metabolic rate when in the differentiated state (3 weeks) as compared to HepG2 cells, and thus may allow the conduct of repeated toxicity testing on cells in culture. In order to evaluate the genetic background of HBG BC2 cells, the expression of selected genes was analyzed in untreated cultures and, in addition, the behavior of HBG BC2 cultures under conditions of repeated treatment was studied with acetaminophen as a test substance and coupled with the use of standard staining techniques to demonstrate toxicity. Results showed that cultures of HBG BC2 cells retained a capacity to undergo apoptosis and proliferation, allowing probable replacement of damaged cells in the culture monolayer. MTT reduction was used to evaluate the toxicity of acetaminophen, acetylsalicylic acid, perhexiline, and propranolol, after both single and repeated (3 times/week for 2 weeks) administration. Under the conditions of repeated treatment, cytotoxicity was observed at lower doses as compared to single administration. In addition, the lowest nontoxic doses were in the same range as plasma concentrations measured in humans under therapeutic use. Our results suggest that the new human hepatoma HBG BC2 cell line is of interest for the evaluation of cell toxicity under conditions of repeated administration. PMID- 12776926 TI - Influence of peracetic acid on adhesion/invasion of Salmonella enterica serotype typhimurium LT2. AB - The influence of peracetic acid (PAA) disinfectant on Salmonella enterica serotype Typhimurium LT2 in sewage effluent was examined by studying its ability to adhere to and invade HeLa cells in vitro. Although the disinfectant produced a decrease of about 5 log units, the bacteria kept their adhesive and invasive abilities. Scanning microscopic observations of the PAA-treated bacteria revealed that PAA caused a loss of external microfilaments and an alteration of membrane structure. Nevertheless, electron-microscopic observations showed that PAA treated bacteria were still able to adhere to and invade HeLa cells despite the fact that the bacteria seemed to have undergone some structural modifications. With confocal microscopy, the use of anti-actin antibody showed that the contact between the bacteria (with or without PAA treatment) and the HeLa cells activated actinopolymerization of the HeLa cell cytoskeleton. PMID- 12776927 TI - Expression of nuclear receptors and apo E secretion during the differentiation of monocytic THP-1 cells into macrophages. AB - The human monocytic THP-1 cell line differentiates into macrophage-like cells that secrete apo E after addition of PMA. Using this model, we studied the time course of apo E transcriptional activation and secretion in relation with the expression of nuclear receptors. Upon treatment with PMA, apo E mRNA and protein secretion were triggered with the concomitant increase of LXRalpha, PPARgamma, and PPARbeta mRNA expression levels. PPARalpha was downregulated, RXRalpha expression was unchanged, and RARalpha and VDR showed only transient increases. FXR and SXR transcripts were not detectable. Specific agonists were used to investigate the functional role of these nuclear receptors upon apo E secretion. The LXRalpha ligands T0901317 and 22(R)-hydroxycholesterol were the most potent apo E inducers, followed by the PPARgamma agonist BRL49653. The PPARalpha agonist Wy14,643 was inactive and 1alpha,25-dihydroxyvitamin D3, 9-cis-retinoic acid and all-trans-retinoic acid decreased apo E secretion. Thus, during PMA-induced THP-1 differentiation, there is a sequential and coordinate regulation of apo E and nuclear receptor transcription. PMID- 12776928 TI - The response of adult rat sertoli cells, immortalized by a temperature-sensitive mutant of SV40, to 1,2-dinitrobenzene, 1,3-dinitrobenzene, 2,4-dinitrotoluene, 3,4-dinitrotoluene, and cadmium. AB - In this study we test the hypothesis that immortalized adult rat Sertoli cells respond to known testicular toxins in a similar manner to Sertoli cells tested in vivo and in primary culture. This cell line was developed by immortalizing adult rat Sertoli cells with the temperature-sensitive mutant of SV40, ts255, such that the cells proliferate at the permissive temperature of 33 degrees C but express differentiated characteristics at the nonpermissive temperature of 40 degrees C. Confluent monolayers, grown at 33 degrees C or 40 degrees C, were exposed to a range of concentrations of dinitrobenzene (DNB) or dinitrotoluene (DNT) isomers or to cadmium chloride. Cellular response was assessed by neutral-red cell viability assay and ultrastructural changes. Cells grown at 40 degrees C were sensitive to lower concentrations of each toxicant than were cells grown at 33 degrees C. 1,2-DNB was more toxic than 1,3-DNB, and 3,4-DNT was more toxic than 2,4-DNT, as judged by the neutral-red cell viability assay. Ultrastructurally, cells treated with 1,2-DNB or 2,4-DNT showed increased numbers of autophagic vesicles compared to controls. Intercellular penetration of ruthenium red demonstrated breached tight junctions in 1,2-DNB and cadmium-treated cells. From these observations, we conclude that this cell line can serve as a model for studying toxic mechanisms in adult Sertoli cells. PMID- 12776929 TI - A fluorescence microplate assay using yopro-1 to measure apoptosis: application to HL60 cells subjected to oxidative stress. AB - A new one-step labeling procedure using the membrane permeant fluorescent probe yopro-1 in association with fluorescence microtitration for the rapid determination of apoptosis is reported. Programmed cell death was induced by the pro-apoptotic agents etoposide and staurosporine, and measured in nonadherent HL60 cells and adherent phorbol 12-myristate 13-acetate (PMA)-treated HL60 cells. Cell viability was controlled by trypan blue exclusion and calcein-AM staining. To confirm results of fluorescence microplate assay, apoptosis was measured by flow cytometry analysis using the same fluorescent probe, and results showed corresponding data between both procedures. Development of apoptosis was confirmed by the presence of PARP (poly(ADP-ribose) polymerase cleavage and nuclear DAPI (4,6-diamidino-2-phenylindole) staining, two well-known methods used to investigate apoptosis. The fluorescence microplate assay was also applied to measure apoptosis in cells exposed to an oxidative stress induced by tert butylhydroperoxide (t-BHP), and results confirmed the potential of the fluorescence microplate assay in measuring events of apoptosis, especially in adherent, cultured, living cells. PMID- 12776930 TI - Bernard Lown and 21 years of International Physicians for Prevention of Nuclear War. PMID- 12776931 TI - The nuclear issue: where do we go from here?. AB - The drive for the elimination of nuclear weapons is going badly and there is currently little support from the general public. The United States Nuclear Posture Review incorporates nuclear capability into conventional war planning. The Stockpile Stewardship Program is designed to maintain nuclear weapon capability. The US is planning an essentially new earth-penetrating nuclear weapon and is prepared to test this in the national interest if thought necessary. These policies could stimulate nuclear proliferation by others, do nothing to deter terrorism, promote persisting polarization of the world, are a clear breach of the Non-Proliferation Treaty and rest world security on a continued balance of terror. A renewed mass campaign to counteract all this, on legal and moral grounds in particular, is urgently needed. IPPNW and kindred organizations must restore sanity in our policies and humanity to our actions. PMID- 12776932 TI - Human security: expanding the scope of public health. AB - Human security is an evolving principle for organizing humanitarian endeavours in the tradition of public health. It places the welfare of people at the core of programmes and policies and is community oriented and preventive. It recognizes the mutual vulnerability of all people and the growing global interdependence that marks the current era. Health is a crucial domain of human security, providing a context within which to build partnerships across disciplines, sectors and agencies. These principles have been demonstrated in field programmes in which healthcare delivery featuring multi-sectoral co-operation across conflict lines has been used to enhance human security. Such programmes can be a model for collaborative action and can create the sustainable community infrastructure that is essential for human security. PMID- 12776933 TI - The responsibility to protect: a new notion of state sovereignty. AB - The recent policies of the United States Administration threaten the entire structure of international law. Widespread concern in the US and elsewhere has been expressed in a different cluster of principles, in particular in the report of the Canadian-based Commission on Intervention and State Sovereignty. Its report states that the primary responsibility of a state is to protect its citizens; such protection includes prevention, reaction and rebuilding. The implications of this for military intervention are discussed; this must be a last resort after all non-military options have failed in response to the actual or probable large-scale loss of life and the objective to prevent further suffering. The action must be authorized by the United Nations Security Council or, if this fails to respond, the General Assembly. A further commission should examine how the international community should respond to states that refuse to comply with international law regarding weapons of mass destruction. PMID- 12776934 TI - 'War on terrorism' and deep culture. AB - This article examines the reasons underlying the massive reaction to the terrorist attacks of 11 September 2001 compared with the lack of response to the many thousands of preventable deaths elsewhere daily. It is suggested that the explanation is the 'deep culture' of the United States based on the myth of the frontiersman. The world is divided into winners and losers, good and bad; non-US citizens are of less value as human beings. The only response allowed by this deep culture is striking back at 'the enemy'. The difficulties with this response are discussed and non-violent alternatives considered. PMID- 12776935 TI - Challenges of war surgery in Mallavi, Sri Lanka. AB - Northern Sri Lanka has been separated by an active front line from the remainder of the country and has lacked basic facilities, including hospital care, because of the war between the government and the Tamil Tigers. This article reports on the resulting situation in one district hospital in the region and on the work of Medecins sans Frontieres in providing specialist hospital care under such difficulties. PMID- 12776936 TI - The Middle East and IPPNW: recent resolutions and declarations. AB - The Israeli-Palestinian conflict is characterized by a cycle of violence from both parties. Their policies will continue to fail until realistic and sincere negotiations begin. This essay describes the activities of affiliates of IPPNW and reports a programme to achieve this. PMID- 12776937 TI - The day we need to remember. AB - The crime of 11 September 2002 is another aspect of the military culture of the last 5,000 years, which is also reflected in the continued conflict in Israel and Palestine. No real attempt has been made to convert the Oslo agreement into a permanent peace settlement, which must include a viable Palestinian state. Nevertheless, all acts of terror must be condemned whatever their motivation. The support of the United States is essential for a Palestinian state to come into being. PMID- 12776938 TI - Will the current crisis lead to the destruction or transformation of the world? PMID- 12776939 TI - World Civil Society Forum. PMID- 12776940 TI - Appeal of the International Committee of the Red Cross on biotechnology, weapons and humanity. PMID- 12776942 TI - The internal structure of bony tissue of a human metopic suture by Soft X-ray. AB - A complete metopic suture was found in the frontal bone of a 79 years old male cadaver. Compact bone matrices were found in the border region of the metopic suture using a Soft X-ray apparatus. The coronal and sagittal sutures were also complete. The appearance of these complete cranial sutures may be due to continuous bone restructuring and resorption in the border region of sutures during brain development of brachy crany. PMID- 12776941 TI - Biological warfare and the MRC. PMID- 12776943 TI - The distribution and morphology of lymphatic vessels on the peritoneal surface of the adult human diaphragm, as revealed by an ink-absorption method. AB - Application of india ink to the peritoneal and pleural surfaces of the adult human diaphragm allowed visualization of the distribution and morphology of the lymphatic vessels by light microscopy and scanning electron microscopy. The diaphragms examined had been fixed and stored in 10% formalin. Numerous lymphatic vessels were stained black with india ink, presenting reticular, radial-meshwork, ladder-like and lacy patterns. They were distributed throughout the entire sternocostal part. Analysis by light and scanning electron microscopy of the areas indicated by india ink revealed the presence of primary lymphatic vessels that formed lymphatic lacunae and stomatal openings to the peritoneal cavity. A layer of secondary collecting lymphatic vessels was located cranially with respect to the layer of primary lymphatic vessels. Thus, the peritoneum had at least two layers of lymphatic vessels. These lymphatic vessels were not tubular vessels but resembled flat cisternae, as has been suggested in the case of the mouse diaphragm. The pleura lacked lymphatic stomata and had no such double layered lymphatic organization. This is the first report that showed distribution and morphology of the lymphatic vessels in the diaphragmatic peritoneum of the formalin-fixed, adult human diaphragm. The method and results in the present study may contribute to morphological analysis of the lymphatic system in the wall of the human body cavity. PMID- 12776944 TI - The aging of human Meissner's corpuscles as evidenced by parallel sectioning. AB - We examined cross-sectional areas of Meissner's corpuscles to evaluate the changes in their morphology and density that occur during the aging process. Our study of 10 tissue specimens from the right index finger pulp of 10 males, showed that both the size and the number of Meissner's corpuscles decrease with age. We claim greater accuracy in our data thanks to the use of a new sectioning method, namely parallel sectioning of the skin. PMID- 12776945 TI - Differences in the axonal compositions of the human mandibular nerve between dentulous and edentulous jaws. AB - We examined the human mandibular nerve to find differences in the composition of nerve fiber axons between dentulous and edentulous jaws Using Goto's modification of Masson-Goldner's method. We discovered that the edentulous jaw did not contain any large size axons, compared with the dentulous jaw. This can be considered as evidence that the larger fibers innervating the periodontal ligament decreased degenerated after tooth loss. PMID- 12776946 TI - Ultrastructure of vascular capillary of lingual filiform papillae in the mole (Mogella wogura wogura). AB - Detailed observation of the structure of filiform papillae (FP) and microvasculature of those papillae in Japanese Azuma mole were described. In the anterior and medial regions, FP was cylinder in shape with two processes. In the posterior region, it had a long, sharp conical shape. The microvascular casts showed two types of hairpin-shaped capillary loops on three regions of the tongue. In the anterior and medial regions, the end of the capillary loops were shaped like a spoon. In contrast, in the posterior region, it was knot-like end of capillary loop. Since the shape of capillary loop was more complex in the anterior and medial regions than that in the posterior region, it was speculated that the spoon-like end of capillary loops of the FP in the anterior and medial regions supply nutrients to the filiform papillary cells and may be related to the movement of the tongue during mastication in Japanese Azuma mole. PMID- 12776947 TI - Beating heart coronary artery bypass grafting: results from 402 patients and the usefulness of gastroepiploic artery composite grafting. AB - OBJECTIVES: We have studied the results of 402 consecutive cases of beating heart coronary artery bypass grafting (CABG) and evaluated the usefulness of gastroepiploic artery (GEA) composite grafts. METHODS: Between March 1993 and August 2001, 402 patients underwent beating heart CABG. They were 321 male and 81 female patients, aged 17 to 88 (mean 66) years. Beating heart CABG was facilitated by mechanical stabilization with a doughnut stabilizer, a newly designed sternal retractor and a new coronary perfusion system. Minimally invasive direct coronary artery bypass (MIDCAB) was performed in 206 patients (the MIDCAB Group), and beating heart CABG with median sternotomy (OPCAB) was performed in 196 patients (the OPCAB Group). RESULTS: Definite off-pump CABG was accomplished in 381 patients. 21 patients (5%) were converted to on-pump beating heart CABG using percutaneous cardiopulmonary system via femoral vessels because of hemodynamic instability. There was 1 operative mortality (0.2%). There was perioperative myocardial infarction in 2 (0.5%), and cerebral infarction in 3 (0.7%). The rate of complete revascularization was 78% in the MIDCAB Group and 97% in the OPCAB Group. The mean number of anastomoses was 1.6 in the MIDCAB Group and 3.3 in the OPCAB Group. The early graft patency was 99.1% in a left internal thoracic artery graft, 97.0% in a right internal thoracic artery graft, 96.5% in GEA, 98.2% in a radial artery graft, and 94.2% in a vein graft. A GEA composite graft was used in 55 of the 168 patients who received GEA grafting. The mean number of anastomoses for the GEA composite graft was 1.6 +/- 0.6 per patient. The graft patency rate was 94.6% (53/56) for GEA and 98.6% (72/73) for the radial artery used as a composite graft. CONCLUSION: A consecutive series of beating heart CABG was performed safely and effectively with a low mortality rate and low morbidity rate. Beating heart CABG could be performed in all patients, and definite off-pump CABG was accomplished in 95% of them. In order to aim for complete revascularization, GEA composite graft was found to be effective since it required a low mean number of 1.6 anastomoses and a satisfactory patency rate at the same time. PMID- 12776948 TI - Changes in hemodynamics in patients who underwent extended mediastinal lymphadenectomy through median sternotomy for primary lung cancer. AB - OBJECTIVES: Extended bilateral mediastinal lymphadenectomy (ND3alpha) through median sternotomy for lung cancer has been reported to show longer survival compared to that after ipsilateral mediastinal lymphadenectomy (ND2a) through posterolateral thoracotomy for node-positive lung cancer. However, a high incidence of morbidity or mortality has also been found. The present prospective study was conducted to clarify the influence of this procedure on the hemodynamics as one of the factors influencing postoperative course. METHODS: Between December 1999 and January 2001, at the Nippon Medical School Hospital, 10 patients underwent pulmonary resection with ND2a through a posterolateral thoracotomy, while another 10 patients underwent pulmonary resection with ND3alpha through median sternotomy. Changes in the hemodynamics were evaluated preoperatively and at 6, 12, 24 and 48 hours postoperatively. RESULTS: Those who underwent ND3alpha showed higher peripheral vascular resistance for 24 hours postoperatively resulting from the longer surgery and increased blood loss, than those who underwent ND2a. The pulmonary capillary wedge pressure was elevated in those who underwent ND3alpha comparing to that in those who underwent ND2a. Stroke index decreased in patients who underwent NDalpha comparing to that in those who underwent ND2a. Even though the right ventricular ejection fraction showed minimal change, both the ventricular stroke work indices were depressed at 48 hours postoperatively in patient who underwent ND3alpha. CONCLUSION: ND3alpha was concluded to have affected the cardiopulmonary circulation, compared to ND2a, and to be more invasive than ND2a. To perform ND3alpha, preoperative patient selection is an important factor to archive the advantage of increased survival by this procedure. PMID- 12776949 TI - Influence of heparin dosage on hemostasis under combined use of Nafamostat mesilate during deep hypothermic circulatory arrest. AB - OBJECTIVE: Nafamostat mesilate (NM) is a guanidine acid derivative which is synthesized in Japan as an antifibrinolytic drug. It has been successfully used to decrease blood loss and blood product requirement in cardiac operations. As there have been some reports that insufficient heparinization might induce the coagulopathic state in aprotinin-treated patients undergoing deep hypothermia and circulatory arrest (DHCA), we have reviewed 16 consecutive patients who underwent total aortic arch replacement using DHCA and the associated use of NM. METHODS: The patients were divided into two groups; a Normal-dosage Group (n = 8) who received 3 mg/kg of heparin, and a Low-dosage Group (n = 8) who received 1.5 mg/kg of heparin. The Celite-activating clotting time was maintained at longer than 1,000 seconds in both groups. Blood loss, transfusion requirements and the volume of postoperative mediastinal tube drainage were compared between the two groups. RESULTS: We observed no adverse effects of NM and no significant difference in transfusion requirements between both groups. There was a significant difference in blood loss during the operation between the two groups (p < 0.05) (Low-dosage Group 1,973 +/- 959 ml vs Normal-dosage Group 1,059 +/- 704 ml). CONCLUSIONS: NM was a safe antifibrinolytic drug. Adequate heparinization was necessary to reduce hemorrhage in patients undergoing aortic arch replacement using DHCA and the associated use of NM. PMID- 12776950 TI - Gene therapy for myocardial angiogenesis: with direct intramuscular gene transfer of naked deoxyribonucleic acid encoding vascular endothelial growth factor and cell transplantation of vascular endothelial growth factor transfected H9c2 myoblast. AB - OBJECTIVES: We studied the therapeutic potential of ex vivo vascular endothelial growth factor (VEGF) gene-transduced H9c2 cell transplantation for myocardial neovascularization. METHODS: The left ventricular free wall of adult Sprague Dawley rats was cryodamaged. Two weeks after, naked plasmid encoding VEGF (VEGF1: 1 1microg/100 microl, VEGF5: 5 microg/100 microl), or VEGF transfected H9c2 myoblasts (0.5 x 10(6) cells/200 microl) were injected into the center of scar tissue. Four weeks after cryoinjury, scar diameters in the each group were measured. Neovascularization in the scar tissue was then quantified histologically. RESULTS: Average scar tissue diameter 4 weeks after cryoinjury was as follows: TE (controls): 6.77 +/- 0.31 mm, H9c2: 5.08 +/- 0.43 mm; VEGF1: 5.90 +/- 0.20 mm; VEGF5: 4.50 +/- 0.24 mm. Scar tissue diameter was smaller in the 3 groups than in the control group (p < 0.05). Capillary density by histological examination increased in naked plasmid injection groups (VEGF1: 1,209.9 +/- 305.3/mm2; VEGF5: 1,072.0 +/- 230.8/mm2) versus controls (708.2 +/- 144.9/mm2, p < 0.05, p < 0.05), and H9c2 cell transplantation group (1,379.4 +/- 391.6/mm2) versus controls (p < 0.05). CONCLUSION: VEGF-transfected H9c2 myoblasts can be implanted and survive in infarcted myocardium. The ability to develop angiogenesis in the H9c2 cell transplantation group was markedly similar to direct myocardial injection in naked plasmid encoding VEGF groups. PMID- 12776951 TI - Aortic intramural hematoma with severe aortic regurgitation. AB - A 71-year-old Japanese woman with severe chest pain was diagnosed with Stanford type A acute aortic dissection. After 3 months of medical treatment, she was operated on under a diagnosis of dissecting aneurysm of the ascending aorta and severe aortic regurgitation. Operative findings showed prolapse of the redundant aortic leaflets and a dilated ascending aorta without intimal tears. Operative and computed tomography findings differed from those of a classical dissection, which was the primary diagnosis of this patient, and were compatible with a diagnosis of aortic intramural hematoma (IMH). Few reports of IMH include concomitant aortic regurgitation. Surgery involved aortic root remodeling and prosthetic graft replacement of the ascending aorta. PMID- 12776952 TI - Aortic root replacement for prosthetic aortic valve detachment without regurgitation and with enlarged Valsalva's sinuses and complete atrioventricular block caused by Takayasu's aortitis. AB - We replaced the aortic root in a 43-year-old woman with Takayasu's aortitis associated with prosthetic aortic valve detachment. The patient's aortic valve had been replaced when she was 31 years old with a mechanical prosthesis to treat aortic regurgitation. Though C-reactive protein was kept almost normal with prednisolone, complete atrioventricular block suddenly appeared 12 years after the first operation. After the implantation of an artificial pacemaker, we closely followed up aortic root status. Aortography and echocardiography showed that the valve moved up and down, probably due to enlargement of the sinuses of Valsalva, without perivalvular leakage. We removed the prosthetic aortic valve, which was partially detached from the aortic valve ring at the right- and non coronary cusps and successfully replaced the aortic root with a mechanical prosthesis inserted into a 26 mm woven graft. Although the postoperative course was uneventful, we closely continue to observe the case and to administer of antiinflammatory medication. PMID- 12776953 TI - Mitral valve surgery in a patient with rheumatoid arthritis being treated with methotrexate. AB - We describe our experience of mitral valve surgery in a 74-year-old man with rheumatoid arthritis (RA). RA had been diagnosed 12 years previously and his symptoms were being controlled by drugs including methotrexate (MTX), which is potentially immuno- and myelo-suppressive. He was admitted for dyspnea, and surgery was indicated for severe mitral insufficiency due to posterior leaflet prolapse. According to the recommendations of orthopedic surgeons, the administration of the MTX was discontinued at two weeks prior to the operation, in which the prolapsed leaflet was excised, repaired, and annuloplasty were performed with a 30 mm prosthetics ring. The patient recovered uneventfully and MTX was resumed one week after surgery. Since MTX has been recently approval for treatment of RA in Japan, Japanese surgeons should pay attention to the appropriate perioperative use of this drug. PMID- 12776954 TI - Acute mitral regurgitation due to total rupture in the anterior papillary muscle after acute myocardial infarction successfully treated by emergency surgery. AB - A 68-year-old female was admitted for sudden onset of chest pain. She received a successful percutaneus coronary intervension for total occlusion in the diagonal artery, but continued to develop progressive heart failure. A chest X-ray showed right pulmonary edema without cardiomegaly, and an echocardiogram revealed massive mitral regurgitation with prolapse in the anterior mitral leaflet due to a rupture in the papillary muscle. An emergency operation was conducted using routine cardiopulmonary bypass. There was complete rupture in the anterior papillary muscle. Mitral valve replacement with posterior mitral leaflet preservation was performed using a size 25 mm Carbomedics prosthetic valve. The postoperative course was uneventful, and she was discharged on postoperative day 29 in New York Heart Association class I. Postoperative pathological findings showed necrosis in the papillary muscle with inflammatory changes. Early diagnosis, prompt medical stabilization, and aggressive surgical intervention are essential to save such a group of patient. PMID- 12776955 TI - Human pulmonary dirofilariasis in a patient whose clinical condition altered during follow-up. AB - Pulmonary dirofilariasis is a rare disease. We have experienced a case which developed pleural effusion while being followed, rendering it difficult to establish the diagnosis. The patient, a 53-year-old woman, had previously undergone two operations; one for uterine cancer and another for breast cancer. She developed a cough in February 2002, and chest computed tomography (CT) scans disclosed a nodular mass in the right lung. A biopsy revealed a fibrous nodule with macrophage aggregation. Pleural effusion was demonstrated on chest CT scans performed in May. As the possibility of malignant tumor could not be ruled out, an operation was undertaken. The nodular lesion showed marked coagulation necrosis, and dead parasites were noted in the vascular lumen. The parasites had the characteristic morphological features of Dirofilaria immitis. Immunological studies produced a positive test result for the anti-dirofilaria immitis antibody; hence a diagnosis of pulmonary dirofilariasis was made. PMID- 12776956 TI - Bilateral internal thoracic artery injury induced by blunt trauma. AB - A case is reported of a 52-year-old man with bilateral internal thoracic artery (ITA) injury induced by blunt trauma. He was a laborer at a construction site who was hit on the anterior chest by a ceiling block. On arrival at the hospital at one hour after the accident, his general condition was good, and the chest computed tomography (CT) showed a slight hematoma at the mediastinum. However, at 4 hours after the trauma, the blood pressure fell, and the CT showed a large anterior mediastinal hematoma with bilateral hemothorax. A median sternotomy revealed bilateral ITA injury at the level of the 3rd intercostal space and incomplete fractures in the sternum and bilateral rib cartilages. Although hemostasis was achieved by ligation of the injured ITA, bleeding re-occurred from the ITA at the level of the 2nd intercostal space, and this was stopped by a second operation. The patient recovered without any further significant complication. The rarity of blunt trauma-induced ITA injury is discussed, including the mechanism, diagnosis, and treatment. PMID- 12776957 TI - Fathers should provide support during childbirth. PMID- 12776958 TI - Brucellar spinal epidural abscess. PMID- 12776959 TI - Labor induction: a decade of change. PMID- 12776960 TI - Should active management of the third stage of labor be routine? PMID- 12776961 TI - Methods for cervical ripening and induction of labor. AB - Induction of labor is common in obstetric practice. According to the most current studies, the rate varies from 9.5 to 33.7 percent of all pregnancies annually. In the absence of a ripe or favorable cervix, a successful vaginal birth is less likely. Therefore, cervical ripening or preparedness for induction should be assessed before a regimen is selected. Assessment is accomplished by calculating a Bishop score. When the Bishop score is less than 6, it is recommended that a cervical ripening agent be used before labor induction. Nonpharmacologic approaches to cervical ripening and labor induction have included herbal compounds, castor oil, hot baths, enemas, sexual intercourse, breast stimulation, acupuncture, acupressure, transcutaneous nerve stimulation, and mechanical and surgical modalities. Of these nonpharmacologic methods, only the mechanical and surgical methods have proven efficacy for cervical ripening or induction of labor. Pharmacologic agents available for cervical ripening and labor induction include prostaglandins, misoprostol, mifepristone, and relaxin. When the Bishop score is favorable, the preferred pharmacologic agent is oxytocin. PMID- 12776962 TI - Congenital toxoplasmosis. AB - Approximately 85 percent of women of childbearing age in the United States are susceptible to acute infection with the protozoan parasite Toxoplasma gondii. Transmission of T. gondii to the fetus can result in serious health problems, including mental retardation, seizures, blindness, and death. Some health problems may not become apparent until the second or third decade of life. An estimated 400 to 4,000 cases of congenital toxoplasmosis occur in the United States each year. Serologic tests are used to diagnose acute T. gondii infection in pregnant women. Because false-positive tests occur frequently, serologic diagnosis must be confirmed at a Toxoplasma reference laboratory before treatment with potentially toxic drugs is considered. In many instances, congenital toxoplasmosis can be prevented by educating pregnant women and other women of childbearing age about not ingesting raw or undercooked meat, using measures to avoid cross-contamination of other foods with raw or undercooked meat, and protecting themselves against exposure to cat litter or contaminated soil. PMID- 12776963 TI - Information from your family doctor. Toxoplasmosis. PMID- 12776964 TI - Diagnostic and therapeutic injection of the hip and knee. AB - Joint injection of the hip and knee regions is a useful diagnostic and therapeutic tool for the family physician. In this article, the injection procedure for the greater trochanteric bursa, the knee joint, the pes anserine bursa, the iliotibial band, and the prepatellar bursa is reviewed. Indications for greater trochanteric bursa injection include acute and chronic inflammation associated with osteoarthritis, rheumatoid arthritis, repetitive use, and other traumatic injuries to the area. For the knee joint, aspiration may be performed to aid in the diagnosis of an unexplained effusion and relieve discomfort caused by an effusion. Injection of the knee can be performed for viscosupplementation or corticosteroid therapy. Indications for corticosteroid injection include advanced osteoarthritis and other inflammatory arthritides, such as gout or calcium pyrophosphate deposition disease. Swelling and tenderness of pes anserine or prepatellar bursae can be relieved with aspiration and corticosteroid injection. Persistent pain and disability from iliotibial band syndrome respond to local injection therapy. The proper technique, choice and quantity of pharmaceuticals, and appropriate follow-up are essential for effective outcomes. PMID- 12776965 TI - Diagnosing the patient with abdominal pain and altered bowel habits: is it irritable bowel syndrome? AB - Diagnosing a patient who presents with abdominal pain and altered bowel habits can be challenging. Although serious organic illnesses can cause these symptoms, irritable bowel syndrome is commonly responsible. It can be difficult to properly evaluate these patients without overusing diagnostic tests and consultation. A practical approach for diagnosing irritable bowel syndrome is suggested, using the Rome II criteria and the presence of alarm symptoms such as weight loss, gastrointestinal bleeding, anemia, fever, or frequent nocturnal symptoms as starting points. If there are no alarm symptoms and the Rome II criteria are not met, it is acceptable to reevaluate the patient at a later date. If there are no alarm symptoms and the Rome II criteria are met, the patient should be categorized on the basis of age: patients 50 years or younger can be evaluated on the basis of predominant symptoms--constipation, diarrhea, or abdominal pain. Patients older than 50 years should be fully evaluated and considered for gastroenterology referral. If alarm symptoms are present, a full evaluation should be performed (and gastroenterology referral considered), regardless of the patient's age. PMID- 12776966 TI - Evaluation of the subfertile man. AB - Infertility affects 15 percent of couples, and 50 percent of male infertility is potentially correctable. Evaluation of the subfertile man requires a complete medical history, physical examination, and laboratory studies. The main purpose of the male evaluation is to identify and treat correctable causes of subfertility. In addition, many men seek an explanation for their condition, which can be discovered during their evaluation. Furthermore, the male fertility evaluation can uncover significant medical and genetic pathology that could affect the patient's health or that of his offspring. Although pregnancies can be achieved without any evaluation other than a semen analysis, this test alone is insufficient to adequately evaluate the male patient. Treatment of correctable male-factor pathology is cost effective, does not increase the risk of multiple births, and can spare the woman invasive procedures and potential complications associated with assisted reproductive technologies. Appropriate evaluation and treatment of the subfertile man are critical in delivering suitable care to the infertile couple. PMID- 12776967 TI - Information from your family doctor. What should I know about male infertility? PMID- 12776968 TI - Screening for type 2 diabetes in adults: recommendations and rationale. PMID- 12776969 TI - Information from your family doctor. Low back pain. PMID- 12776970 TI - Information from your family doctor. When you have a herniated disc. PMID- 12776971 TI - Information from your family doctor. Lumbar spinal canal stenosis: a common cause of back and leg pain. PMID- 12776976 TI - AAP releases policy statement on identifying and treating eating disorders. PMID- 12776977 TI - Psychosocial management of the foot and ankle surgery patient. AB - Currently, many patients undergo surgery when they and their families are not prepared or resilient enough to recover fully, predisposing them to poor outcomes. These poor outcomes lead to missed work, patient depression, chronic pain, litigation, and surgeon frustration. Sometimes these individuals require the surgeon's oversight and are more likely to improve with rapid vocational therapy, physical therapy, and aggressive, continuous chronic-pain management. The foot and ankle surgeon who takes a biopsychosocial multidisciplinary perspective will prescreen his or her patients for positive risk factors and expand his or her intervention long before and after surgery. This strategy of triaging medical cases to differential treatment is not a new concept in medicine. What is novel is the necessity of triaging and prioritizing patients on the basis of the most significant factors that determine successful surgical outcome: psychologic, social, environmental, and historical medical factors . Robert Sternberg of Yale University suggested that three psychologic problem solving strategies are available: (1) I can try to change myself, (2) I can try to change others, or (3) I can try to change the situation. Naturally, the authors of this article encourage applying all three: (1) the caring surgeon is attentive to these issues; (2) the medical community prepares the patient, themselves, and the patient's family; and (3) the environment into which the patient is released is altered to support their rehabilitation. The ABLE Presurgical Assessment Tool and related treatment strategies provide foot and ankle surgeons with an easy-to-use, research-based application to better screen and manage their surgical patients. The goal of this review and assessment tool is not to determine a quantitative level of risk. Instead, the authors hope to facilitate a surgeon's awareness of critical preoperative risk factors and provide a tool to efficiently identify these factors and arrange appropriate treatment as needed. PMID- 12776978 TI - Current trends in preoperative patient evaluation and management for podiatric surgeons. AB - In preparation for elective foot and ankle surgery, the podiatric surgeon often will refer the patient for a preoperative evaluation. Surgeons rely on the input of that consultant to provide a determination as to the operative risk for the patient. This article reviews the fundamental parts of the preoperative evaluation, perioperative patient management, and recent changes and trends within this arena. PMID- 12776979 TI - Preemptive analgesia in foot and ankle surgery. AB - Central neuroplasticity, or changes in CNS processing due to surgical nociception. can amplify postoperative pain. As a result, a hyperalgesic state called wind-up can occur, having debilitating effects on postoperative patients. Preemptive analgesia works to prevent this process and results in a more positive surgical experience. Inhibition of afferent pain pathways by use of local anesthetic blocks, altered perception of pain with opioid use, and inhibition of pain pathways by NMDA receptor antagonists are examples of preemptive analgesia. Using a combination of preemptive modalities and addressing patients' perceptions can aid in interrupting pathologic pain cycles. Positive and modest results have been obtained from animal and human preemptive trials, yet basic pathophysiology demonstrates the validity and importance of preemptive analgesia. Future studies are needed to test effective blockade of afferent input while controlling perception, hyperalgesia, and NMDA receptor activity. The Agency for Health Care Policy and Research now recommends a multifaceted approach to postoperative pain. The goal in pain management is to inhibit destructive pain pathways, maintain intraoperative analgesia, and prevent central sensitization. Preliminary results of multimodal preemptive analgesia trials continue to be promising. PMID- 12776980 TI - Perioperative pain control. AB - There are many options for perioperative pain control available to surgeons. Given these options, adequate levels of analgesia should be achieved and maintained in all surgical patients. Data suggest that analgesia may be improved by combining different analgesic approaches. To avoid high-dose requirements, dose-dependent adverse effects, and potential toxicity associated with reliance on one agent or technique, "balanced" or multimodal analgesic regimens have been advocated. A multimodal recovery program consists of three major components: (1) early mobilization, (2) complete perioperative analgesia, and (3) early oral nutrition. The goal of multimodal programs is to accelerate patient rehabilitation and reduce hospital stays. Balanced multimodal programs are the present and future of perioperative pain control and will enhance patient care. PMID- 12776982 TI - Perioperative management of foot and ankle trauma. AB - This review covers the perioperative management of trauma to the foot and ankle. The goal when treating these injuries is to return the patient to a sensate, plantigrade, painless, and functioning foot and ankle. Depending on the nature of the trauma, realistic outcomes should be established for the patient, family, and surgeon. The importance of early recognition and treatment of foot and ankle injuries has been established and is paramount for the overall recovery of traumatized patient. PMID- 12776981 TI - Deep venous thrombosis risk assessment, incidence, and prophylaxis in foot and ankle surgery. AB - As discussed in this review, DVT and PE are dangerous clinical diagnoses that can occur following foot and ankle surgery. The authors have provided a clinical protocol, a risk assessment tool, and treatment guidelines for this condition that can be applied to the everyday practice of foot and ankle surgeons. Unlike recommendations in previous studies, the authors believe that podiatric and orthopedic surgeons operating on the foot and ankle should evaluate each patient carefully and consider pharmacologic prophylaxis against DVT formation when significant risk factors are present. PMID- 12776983 TI - Postoperative cryotherapy: risks versus benefits of continuous-flow cryotherapy units. AB - Continuous-flow cryotherapy units have been proved to decrease pain, inflammation, swelling, blood loss, and narcotic usage in the postoperative setting. Complications related to cryotherapy are extremely rare (estimated at 0.00225%) but can be devastating. Complications include frostbite, chronic pain, and loss of digits. Caution should be exercised when prescribing continuous-flow cryotherapy units to patients with known risk factors for cold injury. All members involved with care of the patient, including the physician, nursing staff, patient, and patient's family and caregivers, should take an active roll in monitoring the foot for signs and symptoms of cold injury. Continuous-flow cold therapy should be discontinued when capillary refill time is greater than 5 seconds. PMID- 12776984 TI - Extracorporeal shock wave therapy for plantar fasciitis. AB - Shock wave therapy is quickly becoming a mainstay treatment for chronic recalcitrant plantar fasciitis, and many more applications for this therapy may exist. Many advances have been made in regard to this modality of treatment in just a few decades since it was first investigated in the 1950s. Much research has been conducted, revealing the physics and physiologic effect of shock wave therapy. Devices are now approved that produce shock waves for medicinal purposes by one of the three primary methods of shock wave generation. Studies and research are actively being reported and published that demonstrate shock wave therapy success rates comparable to those found in surgical intervention but without the risks inherent in surgery. In conclusion, ESWT has proved to be a viable treatment option for the intervention of chronic recalcitrant plantar fasciitis. PMID- 12776985 TI - Progressive post-traumatic ankle arthrosis treated with total ankle joint replacement: a case review. AB - This article discusses the joint degeneration progression associated with post traumatic arthrosis of the ankle. A representative case study of this debilitating condition was outlined, and treatment with total ankle joint replacement was presented. Although ankle arthrodesis continues to be a standard option following the progression of severe post-traumatic osteoarthritis, total ankle replacement is maturing as a viable option for this condition. PMID- 12776986 TI - Modified surgical approach for retrocalcaneal exostectomy with early return to weight bearing. AB - This procedure is a minimally invasive method of excising a retrocalcaneal exostosis and has been performed at the authors' facility for several years with excellent results and no incidence of Achilles tendon compromise. Postoperative recovery and return to weight bearing are accelerated compared with procedures that reflect the Achilles tendon from its insertion. PMID- 12776987 TI - The FRK/RAK-SHB signaling cascade: a versatile signal-transduction pathway that regulates cell survival, differentiation and proliferation. AB - Recent experiments have unravelled novel signal transduction pathways that involve the SRC homology 2 (SH2) domain adapter protein SHB. SHB is ubiquitously expressed and contains proline rich motifs, a phosphotyrosine binding (PTB) domain, tyrosine phosphorylation sites and an SH2 domain and serves a role in generating signaling complexes in response to tyrosine kinase activation. SHB mediates certain responses in platelet-derived growth factor (PDGF) receptor-, fibroblast growth factor (FGF) receptor-, neural growth factor (NGF) receptor TRKA-, T cell receptor-, interleukin-2 (IL-2) receptor- and focal adhesion kinase (FAK) signaling. Upstream of SHB in some cells lies the SRC-like FYN-Related Kinase FRK/RAK (also named BSK/IYK or GTK). FRK/RAK and SHB exert similar effects when overexpressed in rat phaeochromocytoma (PC12) and beta-cells, where they both induce PC12 cell differentiation and beta-cell proliferation. Furthermore, beta-cell apoptosis is augmented by these proteins under conditions that cause beta-cell degeneration. The FRK/RAK-SHB responses involve FAK and insulin receptor substrates (IRS) -1 and -2. Besides regulating apoptosis, proliferation and differentiation, SHB is also a component of the T cell receptor (TCR) signaling response. In Jurkat T cells, SHB links several signaling components with the TCR and is thus required for IL-2 production. In endothelial cells, SHB both promotes apoptosis under conditions that are anti-angiogenic, but is also required for proper mitogenicity, spreading and tubular morphogenesis. In embryonic stem cells, dominant-negative SHB (R522K) prevents early cavitation of embryoid bodies and reduces differentiation to cells expressing albumin, amylase, insulin and glucagon, suggesting a role of SHB in development. In summary, SHB is a versatile signal transduction molecule that produces diverse biological responses in different cell types under various conditions. SHB operates downstream of GTK in cells that express this kinase. PMID- 12776988 TI - Molecular and genetic mechanisms of obesity: implications for future management. AB - Obesity has become a worldwide public health problem affecting millions of people. A disruption of the balance between energy intake and energy expenditure is believed to be the major cause of obesity. Substantial progress has been made in deciphering the pathogenesis of energy homeostasis over the past few years. The fact that obesity is under strong genetic control has been well established. Human monogenic obesity is rare in large populations, the most common form of obesity is considered to be a polygenic disorder arising from the interaction of multiple genetic and environmental factors. Here, we attempt to briefly review the most recent understanding of molecular mechanisms involved in energy homeostasis and adipogenesis. We discuss the advantages and disadvantages of various approaches commonly used in search for susceptibility genes for obesity. The main results from these genetic studies are summarized, with comments made on the most striking or representative findings. Finally, the implications of the recent advances in the understanding of molecular genetic mechanisms of body weight regulation on prevention and therapeutic intervention of obesity will be discussed. PMID- 12776989 TI - CD4+ T cell responses in the immune control against latent infection by Epstein Barr virus. AB - The human gamma-herpesvirus Epstein-Barr virus establishes latent, life-long infection in more than 95% of the human adult population. Despite its growth transforming capacity, most carriers control EBV associated malignancies efficiently and remain free of EBV+ tumors. It is commonly accepted that lymphoblastoid cells, expressing all EBV latent antigens, are targeted by the immune system and cause tumors only in immune-suppressed individuals. However, immune control of EBV associated malignancies which express only three or one EBV latent antigen is less obvious. Recent studies have addressed the pattern of EBV latent infection in healthy EBV carriers and the identity of EBV derived target antigens for CD4+ T cells. The results suggest that immune surveillance also extends to tumors, which have down-regulated most EBV latent antigens and therefore escape EBV specific immune recognition at least in part. EBV specific immunity that targets these tumors in healthy EBV carriers seems to fail specifically during the development of Hodgkin's disease, nasopharyngeal carcinoma and Burkitt's lymphoma. These three EBV+ tumors appear to subdue EBV immunity against the remaining EBV latent antigens in different ways or profit from the effect of other pathogens on EBV specific immune responses, when they develop in otherwise immune competent individuals. While immune control and immune escape of these so-called spontaneously arising EBV associated malignancies is just beginning to be understood, immune control of persisting EBV infection can serve as a model for tumor immune surveillance in general. PMID- 12776990 TI - Orexigenic/anorexigenic signals in bulimia nervosa. AB - Bulimia nervosa (BN) and Anorexia Nervosa (AN) are currently classified as eating disorders (ED). Both disorders are the product of complex interaction between physiological and psychological and social processes; they are characterized by abnormal eating behavior. However, patients with BN differ from AN in their nutritional state and response of treatment with serotonin-selective reuptake inhibitor (SSRI) as well as frequency of comorbidity of mood and anxiety disorders. Although biological mechanisms of both BN and AN are largely unknown, excess of both feeding-stimulatory and feeding inhibitory signaling in AN have been indicated. This report reviews data that point to the hypothesis that dysregulation of monoaminergic and new peptidergic circuitry controlling food intake and energy expenditure play a major role in the eating behavior of BN. PMID- 12776991 TI - Stroke: molecular mechanisms and potential targets for treatment. AB - Significant advances have been made over the past few years concerning the cellular and molecular events underlying ischemic cell death. The brain succumbs to ischemic injury as a result of loss of metabolic stores, excessive intracellular calcium accumulation, oxidative stress, and potentiation of the inflammatory response. Neurons can also die via necrotic or apoptotic mechanisms, depending on the nature and severity of the insult. While it has been widely held that ischemia is notable for cessation of protein synthesis, brain regions with marginal reduction in blood supply are especially capable of expressing a variety of genes, the functions of many of which are only beginning to be understood. Gene expression is also upregulated upon reperfusion and reoxygenation. As a result, a number of signaling pathways have been identified and are now known to contribute to ischemic progression or, in some cases, attempts at self preservation. This review will focus on the roles of stress genes, apoptosis related genes, and inflammation. Knowledge of such molecular events has fueled interest in developing specific molecular targets with the hope of someday affecting outcome in clinical stroke. PMID- 12776993 TI - New aspects of integrin-mediated leukocyte adhesion in inflammation: regulation by haemostatic factors and bacterial products. AB - Leukocyte recruitment to sites of inflammation, infection or vascular injury is a complex event, depending on a tightly coordinated sequence of leukocyte endothelial- and leukocyte-platelet interactions, which are controlled by the expression and activation of various adhesion receptors and protease systems. The present review will focus on novel aspects of the regulation of integrin dependent leukocyte adhesion by haemostatic factors and bacterial products. In particular, after a short overview of leukocyte recruitment, the review (i) will focus on the crosstalk between haemostatic factors and adhesion molecules with respect to leukocyte extravasation based on the paradigms of the urokinase receptor and high molecular weight kininogen, (ii) will provide information on novel mechanisms for the regulation of leukocyte recruitment by bacterial proteins, on the basis of the anti-inflammatory role of Staphylococcus aureus extracellular adhesive protein and (iii) will draw attention to the junctional adhesion molecules, a novel family of adhesive receptors that are counter receptors for leukocyte integrins and mediate vascular cell interactions. The better understanding of the interactions between vascular cells and particularly of integrin-dependent leukocyte adhesion may lead to the development of novel therapeutical concepts in inflammatory vascular disorders. PMID- 12776992 TI - Regulation of dendritic cell function through Toll-like receptors. AB - Higher animals establish host defense by orchestrating innate and adaptive immunity. This is mediated by professional antigen presenting cells, i.e. dendritic cells (DCs). DCs can incorporate pathogens, produce a variety of cytokines, maturate, and present pathogen-derived peptides to T cells, thereby inducing T cell activation and differentiation. These responses are triggered by microbial recognition through type I transmembrane proteins, Toll-like receptors (TLRs) on DCs. TLRs consist of ten members and each TLR is involved in recognizing a variety of microorganism-derived molecular structures. TLR ligands include cell wall components, proteins, nucleic acids, and synthetic chemical compounds, all of which can activate DCs as immune adjuvants. Each TLR can activate DCs in a similar, but distinct manner. For example, TLRs can be divided into subgroups according to their type I interferon (IFN) inducing ability. TLR2 cannot induce IFN-alpha or IFN-beta, but TLR4 can lead to IFN-beta production. Meanwhile, TLR3, TLR7, and TLR9 can induce both IFN-alpha and IFN-beta. Recent evidences suggest that cytoplamic adapters for TLRs are especially crucial for this functional heterogeneity. Clarifying how DC function is regulated by TLRs should provide us with critical information for manipulating the host defense against a variety of diseases. PMID- 12776994 TI - Examination of the traditional Raman lidar technique. I. Evaluating the temperature-dependent lidar equations. AB - The essential information required for the analysis of Raman lidar water vapor and aerosol data acquired by use of a single laser wavelength is compiled here and in a companion paper [Appl. Opt. 42, 2593 (2003)]. Various details concerning the evaluation of the lidar equations when Raman scattering is measured are covered. These details include the influence of the temperature dependence of both pure rotational and vibrational-rotational Raman scattering on the lidar profile. The full temperature dependence of the Rayleigh-Mie and Raman lidar equations are evaluated by use of a new form of the lidar equation where all the temperature dependence is carried in a single term. The results indicate that, for the range of temperatures encountered in the troposphere, the magnitude of the temperature-dependent effect can reach 10% or more for narrowband Raman water vapor measurements. Also, the calculation of atmospheric transmission, including the effects of depolarization, is examined carefully. Various formulations of Rayleigh cross-section determination commonly used in the lidar field are compared and reveal differences of as much as 5% among the formulations. The influence of multiple scattering on the measurement of aerosol extinction with the Raman lidar technique is considered, as are several photon pulse pileup correction techniques. PMID- 12776995 TI - Examination of the traditional raman lidar technique. II. Evaluating the ratios for water vapor and aerosols. AB - In a companion paper [Appl. Opt. 42, 2571 (2003)] the temperature dependence of Raman scattering and its influence on the Raman and Rayleigh-Mie lidar equations were examined. New forms of the lidar equation were developed to account for this temperature sensitivity.Here those results are used to derive the temperature dependent forms of the equations for the water vapor mixing ratio, the aerosol scattering ratio, the aerosol backscatter coefficient, and the extinction-to backscatter ratio. The error equations are developed, the influence of differential transmission is studied, and several laser sources are considered in the analysis. The results indicate that the temperature functions become significant when narrowband detection is used. Errors of 5% and more can be introduced into the water-vapor mixing ratio calculation at high altitudes, and errors larger than 10% are possible for calculations of aerosol scattering ratio and thus of aerosol backscatter coefficient and of extinction-to-backscatter ratio. PMID- 12776996 TI - Monte Carlo and discrete-ordinate simulations of irradiances in the coupled atmosphere-ocean system. AB - We compare Monte Carlo (MC) and discrete-ordinate radiative-transfer (DISORT) simulations of irradiances in a one-dimensional coupled atmosphere-ocean (CAO) system consisting of horizontal plane-parallel layers. The two models have precisely the same physical basis, including coupling between the atmosphere and the ocean, and we use precisely the same atmospheric and oceanic input parameters for both codes. For a plane atmosphere-ocean interface we find agreement between irradiances obtained with the two codes to within 1%, both in the atmosphere and the ocean. Our tests cover case 1 water, scattering by density fluctuations both in the atmosphere and in the ocean, and scattering by particulate matter represented by a one-parameter Henyey-Greenstein (HG) scattering phase function. The CAO-MC code has an advantage over the CAO-DISORT code in that it can handle surface waves on the atmosphere-ocean interface, but the CAO-DISORT code is computationally much faster. Therefore we use CAO-MC simulations to study the influence of ocean surface waves and propose a way to correct the results of the CAO-DISORT code so as to obtain fast and accurate underwater irradiances in the presence of surface waves. PMID- 12776997 TI - Remote-sensing reflectance of turbid sediment-dominated waters. Reduction of sediment type variations and changing illumination conditions effects by use of reflectance ratios. AB - Variations of sediment type (grain size and refractive index) and changing illumination conditions affect the reflectance signal of coastal waters and limit the accuracy of sediment-concentration estimations from remote-sensing measurements. These effects are analyzed from numerous in situ remote-sensing measurements carried out in the Gironde and Loire Estuaries and then reduced and partly eliminated when reflectance ratios between the near infrared and the visible are considered. These ratios showed high correlation with the sediment concentration. On the basis of the obtained relationships, performing correspondence functions were established that allow an accurate estimation of suspended sediments in the estuaries from Systeme Probatoire d'Observation de la Terre, Landsat, and Sea-Viewing Wide Field-of-View Sensor data, independently of the date of acquisition. PMID- 12776998 TI - Optimal eigenanalysis for the treatment of aerosols in the retrieval of atmospheric composition from transmission measurements. AB - The separation of the individual contributions of aerosol and gases to the total attenuation of radiation through the atmosphere has been the subject of much scientific investigation since remote sensing experiments first began. We describe a new scheme to account for the spectral variation of the aerosol extinction in the inversion of transmission data from occultation measurements. Because the spectral variation of the aerosol extinction is generally unknown,the inversion problem is underdetermined and cannot be solved without a reduction in the number of unknowns in the set of equations used to describe the attenuation at each wavelength. This reduction can be accomplished by a variety of methods, including use of a priori information, the parameterization of the aerosol spectral attenuation, and the specification of the form of the aerosol size distribution. We have developed and implemented a parameterization scheme based on existing empirical and modeled information about the microphysical properties of aerosols. This scheme employs the eigenvectors from an extensive set of simulations to parameterize the aerosol extinction coefficient for incorporation into the inversion algorithm. We examine the accuracy of our method using data sets containing over 24,000 extinction spectra and compare it with that of another scheme that is currently implemented in the Polar Ozone and Aerosol Measurement (POAM) satellite experiment. In simulations using 80 wavelengths in the UV-visible-near-IR spectral range of the Stratospheric Aerosol and Gas Experiment III (SAGE) instrument, we show that, for our optimal parameterization, errors below 1% are observed in 80% of cases, whereas only approximately 20% of all cases are as accurate as this in a quadratic parameterization employing the logarithm of the wavelength. PMID- 12776999 TI - Boundary layer scattering measurements with a charge-coupled device camera lidar. AB - A CCD-based bistatic lidar (CLidar) system has been developed and constructed to measure scattering in the atmospheric boundary layer. The system uses a CCD camera, wide-angle optics, and a laser. Imaging a vertical laser beam from the side allows high-altitude resolution in the boundary layer all the way to the ground. The dynamic range needed for the molecular signal is several orders of magnitude in the standard monostatic method, but only approximately 1 order of magnitude with the CLidar method. Other advantages of the Clidar method include low cost and simplicity. Observations at Mauna Loa Observatory, Hawaii, show excellent agreement with the modeled molecular-scattering signal. The scattering depends on angle (altitude) and the polarization plane of the laser. PMID- 12777000 TI - Use of circular cylinders as surrogates for hexagonal pristine ice crystals in scattering calculations at infrared wavelengths. AB - We investigate the errors associated with the use of circular cylinders as surrogates for hexagonal columns in computing the optical properties of pristine ice crystals at infrared (8-12-microm) wavelengths. The equivalent circular cylinders are specified in terms of volume (V), projected area (A), and volume-to area ratio that are equal to those of the hexagonal columns. We use the T-matrix method to compute the optical properties of the equivalent circular cylinders. We apply the finite-difference time-domain method to compute the optical properties of hexagonal ice columns smaller than 40 microm. For hexagonal columns larger than 40 microm we employ an improved geometric optics method and a stretched scattering potential technique developed in previous studies to calculate the phase function and the extinction (or absorption) efficiency, respectively. The differences between the results for circular cylinders and hexagonal columns are of the order of a few percent. Thus it is quite reasonable to use a circular cylinder geometry as a surrogate for pristine hexagonal ice columns for scattering calculations at infrared (8-12-microm) wavelengths. Although the pristine ice crystals can be approximated as circular cylinders in scattering calculations at infrared wavelengths, it is shown that optical properties of individual aggregates cannot be well approximated by those of individual finite columns or cylinders. PMID- 12777001 TI - Optical bandwidth in coupling: the multicore photonic switch. AB - In the present study, the bandwidth of a photonic switch described previously [Appl. Opt. 37,2296 (1998); 38, 3239 (1999)] is evaluated. First the optical bandwidth is evaluated for coupling between two fiber-core waveguides, in which the cores are embedded within the same cladding. Then the coupling bandwidth is determined for a fiber-core-to-slab-core waveguide, in which the cores are embedded within the same cladding. These bandwidths are then compared and contrasted with the bandwidths of the photonic switch, which consists of two fiber cores and a control waveguide. Two configurations of the photonic switch are considered: one in which the control waveguide is a fiber core and one in which the control waveguide is a slab core. For the photonic switch, the bandwidth characteristics are more complicated than for the coupled pairs, and these characteristics are discussed in detail. PMID- 12777002 TI - Bandwidth estimation for ultra-high-speed lithium niobate modulators. AB - The effects of velocity matching, impedance matching, conductor loss, and dielectric loss on the optical bandwidth of an ultra-high-speed lithium niobate modulator are reported. It is shown that both dielectric loss and impedance matching play a key role for velocity-matched high-speed modulators with low conductor loss. The effects of etch depth, buffer thickness, electrode width, and thegap between the electrodes on device performance are also illustrated. PMID- 12777003 TI - Fiber-optic power limiter based on photothermal defocusing in an optical polymer. AB - We describe the performance of a fiber-optic power-limiting component. The passive device is dynamically responsive to the input signal and has been shown to attenuate continuous-wave power with a dynamic range of up to 9 dB at 150 mW of input power at 1550 nm. The limiting threshold is approximately 30 mW from 1530 to 1565 nm and less than 10 mW at 1430 nm. The device is activated by a photothermal defocusing mechanism in an optical polymer fixed between two expanded core fibers that collimate light through the material. The magitude and threshold of the limiting response is dependent on the absorption properties of the polymer and the size of the gap between the two fiber endfaces. Simple model calculations have been made to predict the limiting response, and they agree reasonably well with the performance of the actual device. PMID- 12777004 TI - Four-channel coarse-wavelength division multiplexing demultiplexer with a modified Mach-Zehnder interferometer configuration on a silicon-on-insulator waveguide. AB - A coarse wavelength division multiplexer is designed on a silicon-on-insulator waveguide using the Mach-Zehnder interferometers with novel multimode interface periodically segmented waveguide couplers and segmented waveguide arms. It is viable for metro and access applications, since it can be inexpensive and provide easy fabrication, compact size, and good output performance. As a design example, the channel spacing of the demultiplexer is chosen to be 24.5 nm for applications to the 10-Gigabit Ethernet. The simulation results show that the wide-passband demultiplexer can have insertion loss less than 2.3 dB and crosstalk better than 18 dB. PMID- 12777005 TI - Implication of rapid thermal annealing-induced cracks on the performance of multiple-quantum-well laser diodes. AB - We investigated the effects of rapid thermal annealing (RTA)-induced cracks on the diode performance fabricated with GaAs-AlGaAs microstructures. These effects were examined and characterized after quantum-well intermixing within an epitaxial structure capped by either SiO2 or SrF2 layers. The results show clearly that the density of surface crackes strongly depends on the atomic interdiffusion between the well and the barrier layers and on the quality of the dielectric caps as well. Moreover, surface-crack correlation with the RTA process an dielectric deposition parameters, and the cracking effects on diode performance were observed and analyzed in detail. The results demonstrate that diode characteristics can be greatly improved by good surface morphology. Most importantly, we explored an effective way of reducing the density of RTA-induced cracks for the dielectrics grown by plasma-enhanced chemical vapor deposition, which was beneficial for dielectric-cap quantum-well disordering. PMID- 12777006 TI - Efficiency dependence on the Tm3+ and Al3+ concentrations for Tm3+-doped silica double-clad fiber lasers. AB - We present measurements of the slope efficiency and the pump power at threshold for a number of Tm3+-doped silica double-clad fiber lasers that incorporate fibers that have a range of Tm3+ concentrations. We obtain a slope efficiency for the approximately 2-microm 3H4 --> 3H6 laser transition that is greater than the Stokes efficiency limit for a Tm3+ concentration as low as 1.3 wt. %. These results indicate that the cross relaxation process, 3F4, 3H6 --> 3H4, 3H4, has a significant effect on the efficiency of the laser despite the relatively short lifetime of the 3F4 energy level. Energy migration of the excitation at the 3F4 level through the process 3F4, 3H6 --> 3H6, 3F4 may be enhancing the cross relaxation mechanism. We also show the importance of reducing the level of clustering of the Tm3+ ion when it is doped into silica by use of appropriate amounts of Al3+ codopant. For Tm3+ concentrations of >1 wt. %, Al3+/Tm3+ concentration ratios of > 10 are recommended forreducing scattering losses, quenching the lifetime, or both. PMID- 12777007 TI - Optical tweezers and fluid characteristics of an optical rotator with slopes on the surface upon which light is incident and a cylindrical body. AB - The characteristics of the optical trapping force, optical torque, and viscous drag force for a newly proposed cylindrical optical rotator are analyzed. The optical trapping force and torque are evaluated by use of a ray optics model for both parallel and focused laser beam illumination. The drag force is calculated from computational fluid dynamics to be the sum of the components of both the pressure and the shearing stress on all the surfaces of the rotator. We analyze the rotation rate by balancing the optical torque with the drag force. A cylindrical optical rotator is expected to rotate at a high speed because of its highly efficient generation of optical torque and its small viscous drag force. PMID- 12777008 TI - 200-mW-average power ultraviolet generation at 0.193 microm in K2Al2B2O7. AB - K2Al2B2O7 has been found to be phase matchable for type-1 sum-frequency generation (SFG) at 0.193 microm by mixing the Nd:YAG laser wavelength at 1.0642 microm and the SFG output of the RbTiOAsO4 optical parametric oscillator tuned at 0.2358 microm. An average power of 200 mW at 10 kHz was obtained in a 7-mm-long crystal. In addition, the Sellmeier equations and the thermo-optic dispersion formula, which predict well the phase-matching conditions and temperature phase matching bandwidths (FWHM) for second-harmonic generation and SFG in the 0.193 0.669-microm range, are presented. PMID- 12777009 TI - Comparison study of CsLiB6O10 and beta-BaB2O4 as nonlinear media for optical parametric oscillators. AB - We compare the relative merits of the new nonlinear materials CsLiB6O10 and beta BaB2O4 for use as gain media in a singly or doubly resonant pulsed optical parametric oscillator pumped by a Q-switched Nd:YAG laser with various parameters. Angle tuning or a wide range can be theoretically stimulated under type I and type II(1) critical phase matching for CsLiB6O10 crystal. The tunable range is 237-2775 nm; the range was extended to the ultraviolet wavelength by fifth-harmonic generation in CsLiB6O10. A comparison of the tunable ranges, resonant thresholds, and conversion efficiencies of CsLiB6O10 and beta-BaB2O4 is obtained by computer simulation. We verify that CsLiB6O10 is an excellent ultraviolet crystal. PMID- 12777010 TI - Optical near-field Raman imaging with subdiffraction resolution. AB - We report optical near-field Raman imaging with subdiffraction resolution (approximately 120 nm) without field enhancement effects. Chemical discrimination on tetracyanoquinodimethane organic thin films showing localized salt complexes is accomplished by detailed Raman maps. Acquisition times that are much shorter than previously reported are due to the high Raman efficiency of the materials and to careful collection and detection of the optical signals in our near-field Raman spectrometer. PMID- 12777011 TI - Synthesis of Hadamard transformers by use of multimode interference optical waveguides. AB - We propose a synthesis method of optical Hadamard transformer using multimode interference (MMI) couplers. By using the signal transfer matrix of 2 x 2, 4 x 4, and 8 x 8 MMI couplers, we show that sum and difference units of input signals can be synthesized. An interchange unit of two signals can also be synthesized. One synthesis method of Hadamard transformers is a combination of only 2 x 2 units, and the other is a combination of N x N(N > or = 4) units as well as 2 x 2 units. The design examples of operation units are shown, and the size and the output power of Hadamard transformers are estimated. PMID- 12777012 TI - Wavelength calibration of spectra measured by the Global Ozone Monitoring Experiment by use of a high-resolution reference spectrum. AB - Earthshine spectra measured by the nadir-viewing Global Ozone Monitoring Experiment (GOME) spectrometer aboard the second European Remote Sensing (ERS-2) Satellite in the range of 240-790 nm are widely used for the retrieval of concentrations and vertical profiles of atmospheric trace gases. For the near real-time delivery of ozone columns and profiles at the Royal Netherlands Meterological Institute, a tailor-made wavelength calibration method was developed. The method use a high-resolution (0.01-nm) solar spectrum as the reference spectrum and applies both a shift and a squeeze to the wavelengths in selected windows to find the optimal wavelength grid per window. This method provides a calibration accuracy of 0.002 nm below and 0.001 nm above 290 nm. The new wavelength calibration method can be used on any wavelength window, for example, to improve the calibration of spectra from the GOME Data Processor. A software package, GomeCal, which performs this recalibration, along with an improved polarization and radiometric correction, has been made and has been released via the World Wide Web. The method can be used for any high-resolution (ir)radiance spectrometer, such as the satellite instruments SCIAMACHY (Scanning Imaging Absorption Spectrometer for Atmospheric Cartography), Ozone Monitoring Instrument, and GOME-2. PMID- 12777013 TI - Remote-sensing reflectance in the Beaufort and Chukchi seas: observations and models. AB - Two semianalytical remote-sensing reflectance models were evaluated and validated by use of bio-optical data collected in the Beaufort and Chukchi seas. Both models were efficient at retrieving chlorophyll concentration, phytoplankton absorption coefficients,and particulate backscattering coefficients. In contrast, they were not accurate in predicting an absorption coefficient for colored dissolved organic matter plus nonpigmented particulates. The poor model performance is attributed to the high variability in the concentrations of these colored materials. A chlorophyll-dependent reflectance model was also assessed, and it proved to be highly successful in reproducing measured reflectance spetra. A four-component, case 2 model with mean absorption spectra for phytoplankton, soluble materials, and nonpigmented particulates was employed in Hydrolight radiative-transfer model simulations. The remote sensing reflectance spectra simulated inthe radiative-transfer model were in excellent agreement with field data. The similarity between the model and the measurement confirms the accuracy of the underlying bio-optical relationships and underscores the utility of modeling for better understanding of the variability of ocean color observations. The latest SeaWiFS algorithm (OC4V4) overestimated chlorophyll by approximately 1.5 fold across most of the observed range of biomass (0.07-9 mg chlorophyll m( 3)). Regionally tuned algorithms explained > 93% of the variability in the surface chlorophyll concentration. PMID- 12777014 TI - Validation of Terra-MODIS phytoplankton chlorophyll fluorescence line height. I. Initial airborne lidar results. AB - The Moderate Resolution Imaging Spectroradiometer (MODIS) aboard the Terra spacecraft contains spectral bands that allow retrieval of solar-induced phytoplankton chlorophyll fluorescence emission radiance. Concurrent airborne laser-induced (and water-Raman normalized) phytoplankton chlorophyll fluorescence data is used to successfully validate the MODIS chlorophyll fluorescence line height (FLH) retrievals within Gulf Stream, continental slope, shelf, and coastal waters of the Middle Atlantic Bight portion of the western North Atlantic Ocean for 11 March 2002. Over the entire approximately 480-km flight line a correlation coefficient of r2 = 0.85 results from regression of the airborne laser data against the MODIS FLH. It is also shown that the MODIS FLH product is not influenced by blue-absorbing chromophoric dissolved organic matter absorption. These regional results strongly suggest that the FLH methodology is equally valid within similar oceanic provinces of global oceans. PMID- 12777015 TI - Chemical imaging sensor and laser beacon. AB - Design and functional aspects of PANSPEC, a panoramic-imaging chemical vapor sensor (PANSPEC is an abbreviation for infrared panoramic-viewing spectroradiometer), were advanced and its optical system reoptimized accordingly. The PANSPEC model unites camera and fused solid-state interferometer and photopolarimeter subsystems. The camera is an eye of the open atmosphere that collects, collimates, and images ambient infrared radiance from a panoramic field of view (FOV). The passive interferometer rapidly measures an infrared-absorbing (or infrared-emitting) chemical cloud traversing the FOV by means of molecular vibrational spectroscopy. The active photopolarimeter system provides a laser beam beacon. This beam carries identification (feature spectra measured by the interferometer) and heading (detector pixels disclosing these feature spectra) information on the hazardous cloud through a binary encryption of Mueller matrix elements. Interferometer and photopolarimeter share a common configuration of photoelastic modulation optics. PANSPEC was optimized for minimum aberrations and maximum resolution of image. The optimized design was evaluated for tolerances in the shaping and mounting of the optical system, stray light, and ghost images at the focal plane given a modulation transfer function metric. PMID- 12777016 TI - Efficient light-scattering calculations for aggregates of large spheres. AB - Calculation of the scattering pattern from aggregates of spheres through the T matrix approach yields high-precision results but at a high-computational cost, especially when the aggregate concerned is large or is composed of large-size spheres. With reference to a specific but representative aggregate, we discuss how and to what extent the computational effort can be reduced but still preserve the qualitative features of the signature of the aggregate concerned. PMID- 12777017 TI - Improving visibility depth in passive underwater imaging by use of polarization. AB - Results are presented that demonstrate the effectiveness of using polarization discrimination to improve visibility when imaging in a scattering medium. The study is motivated by the desire to improve visibility depth in turbid environments, such as the sea. Most previous research in this area has concentrated on the active illumination of objects with polarized light. We consider passive or ambient illumination, such as that deriving from sunlight or a cloudy sky. The basis for the improvements in visibility observed is that single scattering by small particles introduces a significant amount of polarization into light at scattering angles near 90 degrees: This light can then be distinguished from light scattered by an object that remains almost completely unpolarized. Results were obtained from a Monte Carlo simulation and from a small scale experiment in which an object was immersed in a cell filled with polystyrene latex spheres suspended in water. In both cases, the results showed an improvement in contrast and visibility depth for obscuration that was due to Rayleigh particles, but less improvement was obtained for larger scatterers. PMID- 12777018 TI - Estimation of absorption and backscattering coefficients from in situ radiometric measurements: theory and validation in case II waters. AB - A model that relates the coefficients of absorption (a) and backscattering (b(b)) to diffuse attenuation (K(d)), radiance reflectance (R(L)), and the mean cosine for downward irradiance (mu(d)) is presented. Radiance transfer simulations are used to verify the physical validity of the model for a wide range of water column conditions. Analysis of thee radiance transfer simulations suggest that absorption and backscattering can be estimated with average errors of 1% and 3%, respectively, if the value of mu(d) is known with depth. If the input data set is restricted to variables that can be derived from measurements of upward radiance (L(u)) and downward irradiance (E(d)), it is necessary to use approximate values of mu(d). Examination of three different approximation schemes for mu(d) shows that the average error for estimating a and b(b) increases to approximately 13%. We tested the model by using measurements of L(u) and E(d) collected from case II waters off the west coast of Scotland. The resulting estimates of a and b(b) were compared with independent in situ measurements of these parameters. Average errors for the data set were of the order of 10% for both absorption and backscattering. PMID- 12777019 TI - Simulation of infrared scattering from ice aggregates by use of a size-shape distribution of circular ice cylinders. AB - The scalar optical properties (extinction coefficient, mass extinction coefficient, single-scattering albedo, and asymmetry parameter) of a distribution of randomly oriented ice aggregates are simulated generally to well within 4% accuracy by use of a size-shape distribution of randomly oriented circular ice cylinders at wavelengths in the terrestrial window region. The single-scattering properties of the ice aggregates are calculated over the whole size distribution function by the finite-difference time-domain and improved geometric optics methods. The single-scattering properties of the size-shape distribution of circular ice cylinders are calculated by the T-matrix method supplemented by scattering solutions obtained from complex-angular-momentum theory. Moreover, radiative-transfer studies demonstrate that the maximum error in brightness temperature space when the size-shape distribution of circular ice cylinders is used to represent scattering from ice aggregates is only approximately 0.4 K The methodology presented should find wide applicability in remote sensing of ice cloud and parameterization of cirrus cloud scalar optical properties in climate models. PMID- 12777020 TI - Optimization of CH fluorescence diagnostics in flames: range of applicability and improvements with hydrogen addition. AB - This study quantifies the range of premixed flame conditions for which CH fluorescece diagnostics are applicable, and it shows that the CH fluorescence signal can be increased if some of the hydrocarbon fuel is replaced with hydrogen. The CH fluorescence signal is found to be adequate for fuel-air equivalence ratios (phi) as small as 0.85 for both methane-air and propane-air flames. The CH signal increases until a maximum at phi = 1.25 and phi = 1.35 for methane-air and propane-air flames, respectively, and then decreases for richer conditions. A strategy to increase the CH fluorescence signal and decrease interference from soot precursors is proposed by addition of the proper amount of hydrogen to the hydrocarbon fuel. Hydrogen addition reduces the background signal from soot precursors by as much as afactor of 10 and increases the CH fluorescence signal by as much as 80%. The normalized CH fluorescence measurements are compared with computations that utilize GRI-MECH 3.0 chemistry. Sources experimental uncertainties are discussed. PMID- 12777021 TI - Robust monolithic ultraviolet interferometer for the SHIMMER instrument on STPSat 1. AB - We describe the design, fabrication, and testing of a monolithic interferometer consisting entirely of optically contacted fused-silica optical elements that are assembled, adjusted, and permanently bonded in place. The interferometer is part of a spatial heterodyne spectrometer (SHS) [SHIMMER (Spatial Heterodyne Imager for Mesospheric Radicals)] that will be used for near-ultraviolet high-spectral resolution limb imaging of OH solar resonance fluorescence from low Earth orbit aboard the satellite STPSat-1 scheduled for launch in 2006. The stability of the monolith coupled with the relaxed tolerances on optical quality and alignment inherent to SHS make this new instrument extremely robust and especially attractive for applications in harsh environments. PMID- 12777022 TI - Emission following laser-induced breakdown spectroscopy of organic compounds in ambient air. AB - Laser-induced breakdown spectroscopy (LIBS) was applied to nitroaromatic (NC) and polycyclic aromatic hydrocarbon (PAH) samples in ambient air to characterize their resultant emission. Compounds covering various surface, were ablated by use of the second (532-nm) or the fourth (266-nm) harmonic of a nanosecond pulsed Nd:YAG laser. The emission consisted of spectral features related mostly to CNand C2 molecular fragments and to C, H, N, and O atomic fragments. The transitions of the molecular fragments correspond to the CN (B 2sigma+ - X 2sigma+) violet system and the C2 (d 3IIg - a 3IIu) Swan system; the intensity of the former is higher in NCs than in PAHs. The intensity ratios between C2 and CN and between O and N correlate to the molecular structure, suggesting the possibility of distinguishing one chemical class from another and in optimum cases even identifing specific compounds by use of LIBS. PMID- 12777023 TI - Piezoelectric-transducer-based optoelectronic frequency synchronizer for control of pulse delay in a femtosecond passively mode-locked Ti:sapphire laser. AB - We propose a piezoelectric transducer-(PZT-) based optoelectronic frequency synchronizer to control simultaneously change in the repetition rate, the relative pulse delay, and the phase noise of a passively mode-locked femtosecond Ti:sapphire laser with an intracavity saturable Bragg reflector absorber with respect to an electronic frequency reference. An optoelectronic phase-locked-loop based PZT feedback controller with a proportional, integral, and differential (PID) circuit and a tunable voltage regulator is designed to achieve frequency synchronization, phase-noise suppression, and delay-time tuning. When the controlling voltage is tuned from -2.6 to 2.6 V, the maximum pulse-delay range, tuning slope, and tuning resolution of the laser pulse-train are 11.3 ns, 2.3 ps/mV, and 1.2 ps, respectively. Setting the gain constant of the PID circuit at 10 or larger causes the delay-time tuning function to be linearly proportional to the controlling voltage. In the delay-time tuning mode the uncorrelated single side-band phase-noise density of the frequency-synchronized laser is approximately -120 dBc/Hz at an offset frequency of 5 kHz, which is only 7 dBc/Hz higher than that of the electrical frequency reference. The proposed system also supports linear,continuous switching,and programmable control of the delay time of Ti:sapphire laser pulses when they are frequency synchronized to external reference clocks. PMID- 12777024 TI - Amalgamation of the International Society of Refractive Surgery and the American Academy of Ophthalmology. PMID- 12777025 TI - Two-stage laser in situ keratomileusis to correct refractive errors after penetrating keratoplasty. AB - PURPOSE: To evaluate the effect of a two-stage laser in situ keratomileusis (LASIK) procedure on eyes with high astigmatism and/or anisometropia after penetrating keratoplasty. METHODS: Eleven postoperative penetrating keratoplasty eyes were included in a prospective, non-controlled study. All patients had at least 4.00 D of astigmatism and/or at least 3.00 D of anisometropia and were spectacle and contact lens intolerant. Two-stage LASIK was performed; in the first stage a hinged corneal flap 160 microm in thickness and 9 mm in diameter was created. After stabilization of corneal shape (1 to 3 months after keratotomy), the corneal flap was lifted and laser refractive treatment (second stage) was performed. RESULTS: After the first stage, a statistically significant reduction in refractive astigmatism (P<.01) was recorded. In all eyes but one, best spectacle-corrected visual acuity was maintained or improved after the procedure. Three months after the second stage, refractive astigmatism in 8 of 11 eyes (73%) was within +/- 1.00 D, and spherical equivalent refraction in 9 of 11 eyes (82%) was within +/- 1.00 D of intended correction. Preoperative irregular astigmatism persisted in three patients (3 eyes) who could not be corrected within +/- 1.00 D of refractive astigmatism and/or +/- 1.00 D of intended spherical equivalent refraction. In one eye, an interface infiltrate developed shortly after creation of the flap, and resulted in limited melting. CONCLUSIONS: A two-stage LASIK procedure improved visual acuity and refraction in postoperative penetrating keratoplasty eyes with high astigmatism and/or anisometropia. Complications were uncommon but can lead to loss of vision. PMID- 12777026 TI - Proper positioning of the plume evacuator in the VISX Star3 excimer laser minimizes central island formation in patients undergoing laser in situ keratomileusis. AB - PURPOSE: To identify risk factors in a series of patients who developed steep central islands after laser in situ keratomileusis (LASIK). METHODS: We analyzed and compared the refractive and topographic outcome of a study group composed of 83 eyes of 44 patients who underwent LASIK using the VISX Star3 excimer laser with a refraction-matched control group of 83 eyes treated later. The vacuum aspirator of the excimer laser was abnormally positioned during the surgeries performed in the study group. RESULTS: Mean preoperative spherical equivalent refraction in the study group was -6.75 +/- 2.50 D. Four eyes with a mean preoperative spherical equivalent refraction of -9.27 +/- 2.29 D developed steep central islands. Thirty-three (38%) of 83 eyes treated needed retreatment for residual myopia or myopic astigmatism. In the control group, mean preoperative spherical equivalent refraction was -6.76 +/- 2.50 D. Ninety-three percent of eyes were within +/- 1.00 D of target refraction. Five (6.02%) of 83 eyes required retreatment and no eyes developed central islands. CONCLUSION: The abnormally positioned vacuum aspirator coupled with the higher preoperative refractive correction were the likely causative factors for central island formation and the increased incidence of undercorrection in these patients. PMID- 12777027 TI - Higher order ocular aberrations after cycloplegic and non-cycloplegic pupil dilation. AB - PURPOSE: Clinical aberrometry is commonly undertaken with the use of mydriatic agents, however there is no literature available on whether aberrometry results obtained under cycloplegia differ from those obtained without cycloplegia. METHODS: Higher order aberrations were measured over a 6-mm pupil with a Bausch and Lomb Technolas Zywave Aberrometer on the right eyes of 31 young subjects (average age 19.7 +/- 1.7 years; 5 females, 16 males). Two measurement conditions were used for each subject: 1) topical installation of 3 drops 1% cyclopentolate hydrochloride; and 2) topical installation of 1 drop 2.5% phenylephrine hydrochloride, prior to aberrometry measurements. RESULTS: For higher order aberrations (3rd to 5th order), average root mean square (RMS) after phenylephrine measurement (0.3852 microm) was significantly lower than after cyclopentolate (0.4259 microm). A small but statistically significant difference was found between the two conditions for average vertical and horizontal coma and, to a lesser extent, horizontal 5th order aberrations. Repeatability RMS, a measure of test-retest measurement repeatability, was similar for the two conditions at 0.15 microm, and significantly lower than the average RMS for the difference between the two conditions (residual RMS) of 0.22 microm. CONCLUSIONS: The difference between cycloplegic and non-cycloplegic aberration measurements has implications for surgical correction of higher order aberrations. PMID- 12777028 TI - Persistent haze and disorganization of anterior stromal collagen appear unrelated following phototherapeutic keratectomy. AB - PURPOSE: The theoretical effects on corneal transparency induced by changes in collagen fibril packing following phototherapeutic keratectomy were compared to changes in objective measurements of haze. METHODS: Phototherapeutic keratectomy was performed on the right eyes of four young rabbits; left eyes were used as controls. Postoperative slit-lamp measurements of haze were taken at regular intervals up to 19 months. Wounded stromas were studied by synchrotron x-ray diffraction to calculate the average interfibrillar spacing of the collagen fibrils. These data were combined with transmission electron microscope measurements, and the summation of scattered fields method was used to predict the transmission of visible light. RESULTS: Objective measurements of haze were higher than the baseline control throughout the study. Electron micrographs of anterior stroma in 8-month-old wounds displayed irregularly spaced and poorly organized fibrils and x-ray diffraction indicated larger mean interfibrillar spacing compared to the controls. However, the predicted transmission of visible light through the anterior stromal scar tissue was not significantly different than normal. CONCLUSIONS: Following phototherapeutic keratectomy, anterior corneal collagen fibrils were more widely spaced and unevenly organized than in the normal rabbit cornea. However, this did not cause a significant loss of transparency and was therefore unlikely to contribute to haze. PMID- 12777029 TI - Comparison of videokeratographic functional optical zones in conductive keratoplasty and laser in situ keratomileusis for hyperopia. AB - PURPOSE: To compare the videokeratographic functional optical zone of eyes treated with conductive keratoplasty to eyes treated with laser in situ keratomileusis (LASIK) for hyperopia. METHODS: Sixteen eyes treated with conductive keratoplasty for hyperopia were retrospectively evaluated to determine the size of the videokeratographic functional optical zone. The functional optical zone of these eyes was compared to the functional optical zone of 16 eyes that underwent LASIK for hyperopia with the VISX S2 excimer laser, for comparable amounts of hyperopia. The functional optical zone was measured at the edge of central corneal steepening and paracentral flattening on videokeratography 3 to 6 months after surgery. RESULTS: The functional optical zone after surgery measured an average of 5.6 mm horizontally and 5.6 mm vertically in the conductive keratoplasty eyes, and 4.7 mm horizontally and 5.1 mm vertically in the hyperopic LASIK eyes (P<.001 and P<.005). The mean functional optical zone area was 31.1 mm2 in the conductive keratoplasty eyes and 24.6 mm2 in the hyperopic LASIK eyes (P<.001). The functional optical zone created by conductive keratoplasty had more uniform central steepening and less peripheral blending than the functional optical zone created by hyperopic LASIK. CONCLUSION: Conductive keratoplasty was effective at creating central steepening in the cornea. The functional optical zone resulting from conductive keratoplasty was significantly larger than that obtained with hyperopic LASIK using the VISX S2 excimer laser. PMID- 12777030 TI - Ascorbate prophylaxis for corneal haze after photorefractive keratectomy. AB - PURPOSE: To evaluate whether prophylactic systemic ascorbic acid influences the average level of haze and the incidence of late onset corneal haze after photorefractive keratectomy (PRK). METHODS: Two consecutive groups of eyes treated with PRK for myopia with or without astigmatism were retrospectively compared. The patients had been treated similarly, with the exception that systemic ascorbate had been supplied orally in one of the groups. Haze intensity was quantified on a scale from 0 (clear cornea) to 4 (anterior chamber not visible). The diagnostic criterion for late onset corneal haze was a haze grade 2 or higher, occurring 4 to 12 months after surgery. RESULTS: One week, 1, 3, 6, and 12 months after surgery, the group without ascorbate (314 eyes) showed haze of average levels 0.61, 0.51, 0.50, 0.32, 0.10, respectively, and the group with ascorbate (201 eyes) showed haze of average levels 0.38, 0.18, 0.16, 0.09, 0.06, respectively. Comparison of the respective values showed a statistically significant difference between the two groups (P<.01) at 1 week, 1, 3, and 6 months. Late onset corneal haze was observed in 11 eyes in the group without ascorbate, and none was observed in the group with ascorbate (P<.02). CONCLUSION: This retrospective nonrandomized clinical study suggests that oral ascorbic acid supplementation may have a prophylactic effect against haze development after PRK. However, routine prophylactic use of ascorbate can be recommended only after a randomized, prospective clinical trial substantiates its efficacy. PMID- 12777031 TI - Reliability of corneal thickness and endothelial cell density measures. AB - PURPOSE: We evaluated the reliability and agreement between the Orbscan, an ultrasonic pachymeter (Humphrey 855), and the Konan SP 9000-LC in terms of central corneal thickness. The Konan was also used to study the reliability and agreement between endothelial cell density measures. METHODS: Twenty-five normal subjects were examined on two occasions (mean separation = 9 +/- 5 days) by a single examiner using all three instruments for central corneal pachymetry. The Konan Center Method and a manual counting method were performed by two examiners to determine endothelial cell density. Reliability and agreement were assessed by calculating the 95% limits of agreement (LoA) and intraclass correlation coefficients (ICC). RESULTS: For corneal pachymetry test-retest reliability, the 95% limits of agreement were -20 to +17 microm for the ultrasound, -27 to +22 microm for the Konan, and -13 to +13 microm for the Orbscan. There was fair-to good agreement between the pachymeters (intraclass correlation coefficients range = 0.85 to 0.92). For endothelial cell density test-retest reliability, the 95% limits of agreement for the Konan Center Method was -498 to +530, and -482 to +333 cells/mm2 for examiners 1 and 2, respectively. The test-retest 95% limits of agreement for the manual overlaid grid method was -355 to +355, and -535 to +670 cells/mm2 for examiners 1 and 2, respectively. CONCLUSIONS: The reliability and agreement of the Orbscan and Konan corneal pachymeters was good, although the reliability of the Konan for estimating endothelial cell density was fair, at best. PMID- 12777032 TI - Mesopic contrast sensitivity function after laser in situ keratomileusis. AB - PURPOSE: To evaluate contrast sensitivity under mesopic conditions in patients who have undergone uncomplicated bilateral laser in situ keratomileusis (LASIK) for myopia. METHODS: Best-corrected monocular contrast sensitivity was measured with the Stereo Optical F.A.C.T. chart at 1.5, 3, 6, 12, and 18 c/deg in 20 patients (20 left eyes) before and after bilateral LASIK (with the Nidek EC-5000 excimer laser and the Automated Corneal Shaper). Mean preoperative refractive error was -6.40 +/- 1.28 D (range -4.00 to -8.00 D) and postoperative mean refractive error was -0.41 +/- 0.45 D (range -0.75 to +0.50 D). Contrast sensitivity function was measured before and 6 months after surgery using four different chart luminances: 85, 5, 2.5, and 0.1 cd/m2, the first being a photopic level and the rest mesopic. Log contrast sensitivity values at each spatial frequency were used for statistical analysis and normalized values for graphical representation. RESULTS: Contrast sensitivity of postoperative LASIK eyes did not differ from preoperative values at a photopic level (85 cd/m2). However, under mesopic conditions (5 cd/m2 or less) a statistically significant reduction (P<.01) in contrast sensitivity was found at high spatial frequencies (12 and 18 c/deg), although no significant contrast sensitivity differences were observed at low and middle spatial frequencies (P>.01 for 1.5, 3 and 6 c/deg). CONCLUSIONS: LASIK induced significant reductions in contrast sensitivity under mesopic conditions only at high spatial frequencies, even though the photopic contrast sensitivity function was normal. PMID- 12777033 TI - US trends in refractive surgery: 2002 ISRS survey. AB - PURPOSE: To determine the latest trends in refractive surgery in the United States. METHODS: The full U.S. membership of the International Society of Refractive Surgery (ISRS) (approximately 900 members) was mailed the 2002 refractive surgery survey dealing with volumes, types, preferences of refractive surgery performed, and use of emerging technology. RESULTS: Questions regarding RK, AK, PRK, LASIK, LASEK, intracorneal ring segments (ICRS), laser thermal keratoplasty (LTK), conductive keratoplasty (CK), phakic intraocular lenses (PIOL), and clear lens extractions (CLE) were examined in the survey. Procedure preference for low, moderate, and high myopia, and hyperopia, were compared with the results from the surveys of the previous 5 years. Preference for unilateral versus bilateral same-day surgery, laser type, and microkeratome choice were also compared with the survey data from previous years. Incidence and frequency of co management of refractive surgery patients were compared with 1999-2001 data. New questions regarding pupil measurement/documentation, wavefront aberrometry, and custom ablations were incorporated into the 2002 survey. CONCLUSIONS: As refractive surgery grows in the mainstream of ophthalmology, trends and changes in the United States continue to be elucidated by this professional organization survey. LASIK continues to dominate for refractive errors between -10.00 to +3.00 D. LASIK, LASEK, CLE, PIOL, and CK appear to have bright futures, whereas, RK, ICR, and LTK are on the decline. VISX continues to be utilized 2:1 over all other lasers combined, and instrumentation pupillometry is preferred 2:1 over pupil gauge cards. Currently, wavefront aberrometry and custom ablations are minimally employed but appear poised to be the wave of the future. PMID- 12777034 TI - Fungal keratitis after photorefractive keratectomy: delayed diagnosis and treatment in a co-managed setting. AB - PURPOSE: To report a case of fungal keratitis following photorefractive keratectomy (PRK) in a co-managed setting. METHODS: A 35-year-old man with a preoperative refraction of -13.00 +3.75 x 18 degrees OD, and -12.50 +2.50 x 70 degrees OS underwent bilateral simultaneous PRK. On postoperative day 16, the patient presented with complaints of decreased vision and foreign body sensation. Examination by the co-managing optometrist revealed a visual acuity of 20/200, and a central corneal ulcer. Cultures were not taken and the patient was started on topical ofloxacin and prednisolone acetate 1% every hour. The patient was instructed to follow-up with another optometrist closer to the patient's home. Tobradex (tobramycin 0.3% combined with dexamethasone 0.1%) was added to the treatment regimen on postoperative day 23, but the ulcer continued to worsen. The patient was then referred to an ophthalmologist where corneal cultures were performed, but came back negative. RESULTS: Despite treatment with fortified antibiotics, the ulcer perforated, requiring penetrating keratoplasty. Bacterial and fungal cultures were negative. Pathology examination of the specimen revealed fungal hyphae. CONCLUSIONS: Although co-management of refractive patients may be a common practice, it is not without risks. In this case, delayed diagnosis and inappropriate treatment resulted in a poor final outcome. PMID- 12777035 TI - Experimental imaging of intracorneal ring segments with optical coherence tomography. AB - PURPOSE: To investigate the imaging of corneal structures with optical coherence tomography (OCT) after implantation of intracorneal ring segments (ICRS). METHODS: In an experimental study with six porcine eyes, qualitative and quantitative imaging with corneal OCT using a wavelength of 1310 nm after implantation of ICRS was performed. The optical results were compared with light microscopy of the histological sections. RESULTS: In corneal OCT, the ICRS revealed marked hyporeflective intrastromal areas, which correlated well with macroscopic and microscopic findings. Corneal OCT enabled precise images of the incision depth for the implantation of ICRS, and the exact intrastromal segment position. CONCLUSIONS: Noncontact slit lamp-adapted corneal optical coherence tomography could be employed to clinically monitor corneal changes after implantation of ICRS, evaluate the depth of the segments to correlate refractive changes, and quantify the stromal wound healing response. PMID- 12777036 TI - Intraoperative optical coherence pachymetry during laser in situ keratomileusis- first clinical experience. AB - PURPOSE: To investigate intraoperative optical coherence pachymetry during laser in situ keratomileusis (LASIK). METHODS: In an initial clinical evaluation, three patients with myopia and myopic astigmatism were studied. Corneal thickness was assessed with optical pachymetry based on low-coherence interferometry during LASIK. RESULTS: The attempted mean spherical equivalent refraction was -5.70 +/- 2.00 D with a mean calculated stromal ablation depth of 95 +/- 18 microm. Intraoperative optical coherence pachymetry was reproducible in all patients during the different stages of LASIK, demonstrating a mean flap thickness of 141 +/- 30 microm with a residual corneal stroma of 274 +/- 24 microm at the end of the laser ablation. The immediate postoperative corneal thickness revealed marked swelling. CONCLUSIONS: This initial clinical evaluation proved that intraoperative optical coherence pachymetry may be an important safety feature for monitoring flap and residual stromal thickness during LASIK. It may be particularly helpful in the effort to avoid iatrogenic corneal ectasia in patients with thin corneas, higher refractive corrections, and LASIK enhancements. PMID- 12777037 TI - Central corneal thickness in Indian patients undergoing LASIK for myopia. PMID- 12777038 TI - Bilateral Mycobacterium chelonae keratitis following LASIK. PMID- 12777039 TI - Influence of medical and natural mydriasis on higher order aberrations of the eye. PMID- 12777040 TI - Clinical outcome of clear lens extraction in high myopia after epikeratoplasty lenticule removal. PMID- 12777042 TI - Tobacco bZIP factor TGA10 is a novel member of the TGA family of transcription factors. AB - TGA factors constitute a family of conserved plant bZ1P transcription factors that regulate transcription from as-1-like elements in response to plant signalling molecules salicylic acid (SA), methyl jasmonate (MJ) and auxin. Based on sequence similarities, two subclasses of TGA factors have been identified before in tobacco: class I factors (TGA1a and PG13) are preferentially expressed in root tip meristems, whereas class II factors (TGA2.1 and TGA2.2) are found in leaves and in roots. Here we describe a novel member of the tobacco TGA family (TGA10), which defines a distinct subclass of its own. TGA10 mRNA and TGA10 protein were found in roots but not in leaves of mature tobacco plants. TGA10 binds specifically to the as-1 element, interacts with TGA2.2, and activates transcription in yeast. When ectopically expressed in leaves, TGA10 enhanced SA-, auxin- and MJ-inducibility of target gene Nt103, which responds in the same manner to enhanced levels of TGA2.2. This indicates that TGA10, albeit normally not present in leaves, can interact with the leaf regulatory network controlling transcription from as-1-containing promoters. However, Nt103 expression was not affected in roots of TGA10-over-expressing plants, implying the existence of root specific mechanisms which do not allow a positive effect of increased TGA10 levels on target gene expression. PMID- 12777041 TI - Differential expression of genes encoding calmodulin-binding proteins in response to bacterial pathogens and inducers of defense responses. AB - Calmodulin (CaM) plays an important role in sensing and transducing changes in cellular Ca2+ concentration in response to several biotic and abiotic stresses. Although CaM is implicated in plant-pathogen interactions, its molecular targets and their role in defense signaling pathway(s) are poorly understood. To elucidate the signaling pathways that link CaM to defense responses, we screened a cDNA library constructed from bean leaves undergoing a hypersensitive response (HR) with radiolabeled CaM isoforms. A total of 26 putative CBPs were identified. Sequencing of the cDNAs revealed that they represent 8 different genes. They are homologues of previously identified CaM-binding proteins (CBPs) in other systems. However, some CBPs are novel members of known CBP families. The proteins encoded by these clones bound CaM in a Ca2+-dependent manner. To determine if these CBPs are involved in plant defense responses, we analyzed their expression in bean leaves inoculated with compatible, incompatible and nonpathogenic bacterial strains. Expression of three CBPs including an isoform of cyclic nucleotide-gated channels (PvCNGC-A) and two hypothetical proteins (PvCBP60-C and PvCBP60-D) was induced whereas the expression of two other isoforms of CNGCs (PvCNGC-B and PvCNGC-C) was repressed in response to incompatible pathogens. The expression of the rest, a small auxin up RNA (PvSAUR1) and two hypothetical proteins (PvCBP60-A and PvCBP60-B), was not changed. The expression of most of the pathogen-regulated genes was also affected by salicylic acid, jasmonic acid, hydrogen peroxide and a fungal elicitor, which are known to induce defense responses. Our results strongly suggest that at least five bean CBPs are involved in plant defense responses. PMID- 12777043 TI - A novel oxidative stress-inducible peroxidase promoter from sweetpotato: molecular cloning and characterization in transgenic tobacco plants and cultured cells. AB - A strong oxidative stress-inducible peroxidase (POD) promoter was cloned from sweetpotato (Ipomoea batatas) and characterized in transgenic tobacco plants and cultured cells in terms of environmental stress. A POD genomic clone (referred to as SWPA2) consisted of 1824 bp of sequence upstream of the translation start site, two introns (743 bp and 97 bp), and a 1073 bp coding region. SWPA2 had previously been found to encode an anionic POD which was highly expressed in response to oxidative stress. The SWPA2 promoter contained several cis-element sequences implicated in oxidative stress such as GCN-4, AP-1, HSTF, SP-1 reported in animal cells and a plant specific G-box. Employing a transient expression assay in tobacco protoplasts, with five different 5'-deletion mutants of the SWPA2 promoter fused to the beta-glucuronidase (GUS) reporter gene, the 1314 bp mutant deletion mutant showed about 30 times higher GUS expression than the CaMV 35S promoter. The expression of GUS activity in transgenic tobacco plants under the control of the -1314 SWPA2 promoter was strongly induced in response to environmental stresses including hydrogen peroxide, wounding and UV treatment. Furthermore, GUS activity in suspension cultures of transgenic cells derived from transgenic tobacco leaves containing the -1314 bp SWPA2 promoter-GUS fusion was strongly expressed after 15 days of subculture compared to other deletion mutants. We anticipate that the -1314 bp SWPA2 promoter will be biotechnologically useful for the development of transgenic plants with enhanced tolerance to environmental stress and particularly transgenic cell lines engineered to produce key pharmaceutical proteins. PMID- 12777044 TI - Sporamin-mediated resistance to beet cyst nematodes (Heterodera schachtii Schm.) is dependent on trypsin inhibitory activity in sugar beet (Beta vulgaris L.) hairy roots. AB - Sporamin, a sweet potato tuberous storage protein, is a Kunitz-type trypsin inhibitor. Its capability of conferring insect-resistance on transgenic tobacco and cauliflower has been confirmed. To test its potential as an anti-feedant for the beet cyst nematode (Heterodera schachtii Schm.), the sporamin gene SpTI-1 was introduced into sugar beet (Beta vulgaris L.) by Agrobacterium rhizogenes mediated transformation. Twelve different hairy root clones expressing sporamin were selected for studying nematode development. Of these, 8 hairy root clones were found to show significant efficiency in inhibiting the growth and development of the female nematodes whereas 4 root clones did not show any inhibitory effects even though the SpTI-1 gene was regularly expressed in all of the tested hairy roots as revealed by northern and western analyses. Inhibition of nematode development correlated with trypsin inhibitor activity but not with the amount of sporamin expressed in hairy roots. These data demonstrate that the trypsin inhibitor activity is the critical factor for inhibiting growth and development of cyst nematodes in sugar beet hairy roots expressing the sporamin gene. Hence, the sweet potato sporamin can be used as a new and effective anti feedant for controlling cyst nematodes offering an alternative strategy for establishing nematode resistance in crops. PMID- 12777046 TI - The age of the Arabidopsis thaliana genome duplication. AB - We estimate the timing of the Arabidopsis thaliana whole-genome duplication by means of phylogenetic and statistical analysis, and propose two possible scenarios for the duplication. The first one, based on the assumption that the duplicated segments diverged from an autotetraploid form, places the duplication at about 38 million years ago, after the Arabidopsis lineage diverged from that of soybean (Glycine max) and before it diverged from its sister genus, Brassica. The second scenario assumes that the ancestor was allotetraploid, and suggests that the duplication is younger than 38 million years and may have contributed to the Arabidopsis-Brassica divergence. In each case, our estimate places the age of the genome duplication as significantly younger than previously reported. PMID- 12777045 TI - Pectate lyase gene expression and enzyme activity in ripening banana fruit. AB - Two distinct cDNA clones showing sequence homology to higher-plant pectate lyase (Pel) genes were isolated from ripening banana fruits. The transcripts were detected only in fruit tissue and both were strongly ripening-related. Yeast transformation with the most highly expressed Pel clone produced a recombinant protein with pectate lyase activity, demonstrating that this sequence was likely to encode a pectate lyase protein in planta. An assay developed for measuring the action of the endogenous enzyme from banana pulp tissue revealed a significant increase in calcium-dependent pectate lyase activity during ripening. The enhanced levels of enzyme activity corresponded with an increase in soluble polyuronides from banana pulp. PMID- 12777047 TI - Expression of AODEF, a B-functional MADS-box gene, in stamens and inner tepals of the dioecious species Asparagus officinalis L. AB - Garden asparagus (Asparagus officinalis L.) is a dioecious species with male and female flowers on separate unisexual individuals. Since B- and C-functional MADS box genes specify male and female reproductive organs, it is important to characterize these genes to clarify the mechanism of sex determination in monoecious and dioecious species. In this study, we isolated and characterized AODEF gene, a B-functional gene in the development of male and female flowers of A. officinalis. Southern hybridization identified a single copy of AODEF gene in asparagus genome. Northern blot analysis showed that this gene was specifically expressed in flower buds and not in vegetative tissues. In situ hybridization showed that during early hermaphrodite stages, AODEF gene was expressed in the inner tepal and stamen whorls (whorls 2 and 3, respectively), but not in the outer tepals (whorl 1), in both male and female flowers. In late unisexual developmental stages, the expression of AODEF gene was still detected in the inner tepals and stamens of male flowers, but the expression was reduced in whorls 2 and 3 of female flowers. Our results suggest that AODEF gene is probably not involved in tepal development in asparagus and that the expression of AODEF gene is probably controlled directly or indirectly by sex determination gene in the late developmental stages. PMID- 12777048 TI - The fuc1 gene product (20 kDa FUC1) of Pisum sativum has no alpha-L-fucosidase activity. AB - An alpha-L-fucosidase purified from pea (Pisum sativum L. cv Alaska) epicotyl was previously described as a cell wall enzyme of 20 kDa that hydrolyses terminal alpha-L-fucosidic linkages from oligosaccharide fragments of xyloglucan. cDNA and genomic copies were further isolated and sequenced. The predicted product of the cDNA and the genomic clone (fuc1), was a 20 kDa protein containing a signal peptide and five cysteines. This was the first alpha-L-fucosidase gene to be cloned in plants but its fucosidase activity has not been demonstrated. Here, our biochemical and immuno analyses suggest that fuc1 does not encode an alpha-L fucosidase. Pea fuc1 expressed in Escherichia coli, insect cells and Arabidopsis thaliana produced recombinant proteins without alpha-L-fucosidase activity. Pea plants had endogenous alpha-L-fucosidase activity, but the enzyme was not recognised by an antibody produced against recombinant FUC1 protein expressed in E. coli. In contrast, the antibody immunoprecipitated a 20 kDa protein which was inactive. By chromatographic analysis of pea protein extracts, we separated alpha L-fucosidase-active fractions from the 20 kDa protein fractions. We conclude that the alpha-L-fucosidase activity is not attributable to the 20 kDa FUC1 protein. A new function for fuc1 gene product, now named PIP20 (for protease inhibitor from pea) is proposed. PMID- 12777049 TI - C-terminal extension of phaseolin with a short methionine-rich sequence can inhibit trimerisation and result in high instability. AB - In an attempt to increase the content in essential amino acids methionine and tryptophan of the trimeric storage protein phaseolin, we fused a Met- and Trp rich sequence to the C-terminus of a phaseolin variant lacking its vacuolar sorting signal, with the aim to target the protein for secretion and accumulation into the apoplast. The fate of the mutant protein, denominated Y3, was studied in transiently transfected tobacco protoplasts. We report that the presence of the additional sequence causes structural defects which inhibit trimerization and lead to partial aggregation of Y3. The protein interacts with the ER chaperone BiP prior to being degraded very rapidly, in a process that does not require vesicular transport from the ER. The rate of degradation of Y3 is higher than that observed for another assembly defective mutant of phaseolin, delta360, which remains monomeric and does not aggregate. This indicates that the plant ER quality control machinery can dispose of defective proteins with different kinetics and perhaps mechanisms, depending on the nature of their defect. PMID- 12777050 TI - Jasmonate biosynthesis and the allene oxide cyclase family of Arabidopsis thaliana. AB - In biosynthesis of octadecanoids and jasmonate (JA), the naturally occurring enantiomer is established in a step catalysed by the gene cloned recently from tomato as a single-copy gene (Ziegler et al., 2000). Based on sequence homology, four full-length cDNAs were isolated from Arabidopsis thaliana ecotype Columbia coding for proteins with AOC activity. The expression of AOC genes was transiently and differentially up-regulated upon wounding both locally and systemically and was induced by JA treatment. In contrast, AOC protein appeared at constitutively high basal levels and was slightly increased by the treatments. Immunohistochemical analyses revealed abundant occurrence of AOC protein as well as of the preceding enzymes in octadecanoid biosynthesis, lipoxygenase (LOX) and allene oxide synthase (AOS), in fully developed tissues, but much less so in 7 day old leaf tissues. Metabolic profiling data of free and esterified polyunsaturated fatty acids and lipid peroxidation products including JA and octadecanoids in wild-type leaves and the jasmonate-deficient mutant OPDA reductase 3 (opr3) revealed preferential activity of the AOS branch within the LOX pathway. 13-LOX products occurred predominantly as esterified derivatives, and all 13-hydroperoxy derivatives were below the detection limits. There was a constitutive high level of free 12-oxo-phytodienoic acid (OPDA) in untreated wild type and opr3 leaves, but an undetectable-expression of AOC. Upon wounding opr3 leaves exhibited only low expression of AOC, wounded wild-type leaves, however, accumulated JA and AOC mRNA. These and further data suggest regulation of JA biosynthesis by OPDA compartmentalization and a positive feedback by JA during leaf development. PMID- 12777051 TI - Ferredoxin from sweet pepper (Capsicum annuum L.) intensifying harpin(pss) mediated hypersensitive response shows an enhanced production of active oxygen species (AOS). AB - The hypersensitive response (HR) is a form of cell death associated with plant resistance to pathogen infection. Harpin(pss), an elicitor from the bacterium Pseudomonas syringae pv. syringae, induces a HR in non-host plants. Previously, we reported an amphipathic protein from sweet pepper interfering with harpin(pss) mediated HR. In this report, we isolated and characterized a cDNA clone encoded that amphipathic protein from sweet pepper. This protein is designated as PFLP (plant ferredoxin-like protein) by virtue of its high homology with plant ferredoxin protein containing an N-terminal signal peptide responsible for chloroplast targeting and a putative 2Fe-2S domain responsible for redox activity. Recombinant PFLP obtained from Escherichia coli was able to significantly increase active oxygen species (AOS) generation when mixed with harpin(pss) in tobacco suspension cells. It also showed enhanced HR when co infiltrated with harpin(pss) in tobacco leaves. We used a transgenic tobacco suspension cells system that constitutively expresses the Pflp gene driven by the CaMV 35S promoter to study the function of PFLP in enhancing harpin(pss)-mediated hypersensitive cell death in vivo. In response to harpin(pss), suspension cells derived from Pflp transgenic tobacco showed a significant increase both in the generation of AOS and in cell death as compared to the wild type. AOS inhibitors diphenylene iodonium chloride (DPI) and lanthanum chlorate (LaCl3) were used to study the involvement of AOS in harpin(pss)-induced cell death. Our results demonstrate enhanced generation of AOS is necessary to cause enhanced hypersensitive cell death in Pflp transgenic tobacco cells and it is plasma membrane-bound NADPH-oxidase-dependent. Sub-cellular localization studies showed that PFLP is present in the cytoplasm and chloroplast of Pflp transgenic tobacco cells, but only in the chloroplast, not in the cytoplasm, of wild-type tobacco cells. It is possible that PFLP can change the redox state of the cell upon harpin(pss) inoculation to increase AOS generation and hypersensitive cell death. Overall, this study will provide a new insight in the functional properties of ferredoxin in hypersensitive cell death. PMID- 12777053 TI - Oscillation of mRNA level and activity of granule-bound starch synthase I in Arabidopsis leaves during the day/night cycle. AB - Granule-bound starch synthase (GBSSI) is one of the most extensively studied enzymes of the starch synthesis pathway and its role in the synthesis of amylose has been well established. However, few studies have been carried out to characterize the regulation of GBSSI gene. Regulation of starch synthesis genes is especially interesting in photosynthetic tissues, where starch is subjected to a periodical alternation of synthesis and degradation during the day/night cycle. In this report we show a circadian oscillation of GBSSI mRNA levels in leaves of Arabidopsis during the day/night cycle, and provide evidence that GBSSI expression is controlled by the transcription factors CCA1 and LHY. Over expression of both CCA1 and LHY genes causes the elimination of GBSSI mRNA oscillation. Binding shift assays indicate that this control may be exerted through a direct interaction of those regulatory proteins with the GBSSI promoter. Oscillation is not observed on the GBSSI protein levels, which remains constant along the cycle. However, GBSSI activity shows a clear oscillation with a period of 24 h that is altered in transgenic plants over-expressing CCA1. Possible mechanisms controlling GBSSI activity during the day/night cycle are discussed. PMID- 12777052 TI - Genome organization in Arabidopsis thaliana: a survey for genes involved in isoprenoid and chlorophyll metabolism. AB - The isoprenoid biosynthetic pathway provides intermediates for the synthesis of a multitude of natural products which serve numerous biochemical functions in plants: sterols (isoprenoids with a C30 backbone) are essential components of membranes; carotenoids (C40) and chlorophylls (which contain a C20 isoprenoid side-chain) act as photosynthetic pigments; plastoquinone, phylloquinone and ubiquinone (all of which contain long isoprenoid side-chains) participate in electron transport chains; gibberellins (C20), brassinosteroids (C30) and abscisic acid (C15) are phytohormones derived from isoprenoid intermediates; prenylation of proteins (with C15 or C20 isoprenoid moieties) may mediate subcellular targeting and regulation of activity; and several monoterpenes (C10), sesquiterpenes (C15) and diterpenes (C20) have been demonstrated to be involved in plant defense. Here we present a comprehensive analysis of genes coding for enzymes involved in the metabolism of isoprenoid-derived compounds in Arabidopsis thaliana. By combining homology and sequence motif searches with knowledge regarding the phylogenetic distribution of pathways of isoprenoid metabolism across species, candidate genes for these pathways in A. thaliana were obtained. A detailed analysis of the vicinity of chromosome loci for genes of isoprenoid metabolism in A. thaliana provided evidence for the clustering of genes involved in common pathways. Multiple sequence alignments were used to estimate the number of genes in gene families and sequence relationship trees were utilized to classify their individual members. The integration of all these datasets allows the generation of a knowledge-based metabolic map of isoprenoid metabolic pathways in A. thaliana and provides a substantial improvement of the currently available gene annotation. PMID- 12777054 TI - Characterization, expression and phylogenetic study of R2R3-MYB genes in orchid. AB - cDNA fragments representing 21 R2R3-MYB genes were isolated by RT-PCR from the Dendrobium orchid hybrid Woo Leng. Six full-length cDNA clones were obtained from a flower cDNA library, four of which, DwMYB1, DwMYB2, DwMYB8 and DwMYB10, represent typical plant R2R3-MYB genes. The conceptual DwMYB4 protein is truncated at the C-terminal region and contains the R2 repeat and the N-terminal half of the R3 repeat (R2R3'). DwMYB4 expression is restricted to flowers. DwMYB9 contains an 8 amino acid N-terminal deletion in the R2 repeat (R2'R3) and is expressed at high levels in mature flower and inflorescence, but at very low levels in young flower buds. DwMYB8 and DwMYB10 show similar expression patterns and share very high sequence similarity in the N-terminal part of the MYB domain. Analysis of amino acid substitution indicated that the pattern and type of substitution between Arabidopsis and maize are quite different. Maize may have more conserved substitution in the MYB(BRH) domain than Arabidopsis. PMID- 12777056 TI - Double-stranded RNA replicons associated with chloroplasts of a green alga, Bryopsis cinicola. AB - Double-stranded RNAs (dsRNAs) associated with chloroplasts and mitochondria have been found in the coenocytic green alga Bryopsis cinicola. In this study we report molecular properties of the four chloroplast-associated dsRNAs (BDRC1 to BDRC4). The longest dsRNA molecule (BDRC1) was sequenced entirely (1959 bp) and a single large ORF of 1722 bp was found within it. Database searches revealed similarities between the deduced amino acid sequence of this ORF and RNA dependent RNA polymerase (RdRp) sequences from several RNA viruses. The most similar sequence in the database was the RdRp of beet cryptic virus 3. Phylogenetic analysis revealed that the RdRp-like sequence of BDRC1 can be placed in the Partitiviridae clade. To detect autonomous replication of these dsRNAs, RdRp assays were carried out with actinomycin D, which is an inhibitor of DNA dependent RNA synthesis. Incorporation of [alpha-32P]UTP was detected specifically in the chloroplast and mitochondrial dsRNAs, indicating that both the chloroplast dsRNAs (BDRCs) and the mitochondrial dsRNA (BDRM) of B. cinicola are RNA replicons. The green alga B. cinicola harbors different dsRNA replicons in its chloroplasts and mitochondria. PMID- 12777055 TI - A new Arabidopsis thaliana mutant deficient in the expression of O methyltransferase impacts lignins and sinapoyl esters. AB - A promoter-trap screen allowed us to identify an Arabidopsis line expressing GUS in the root vascular tissues. T-DNA border sequencing showed that the line was mutated in the caffeic acid O-methyltransferase 1 gene (AtOMT1) and therefore deficient in OMT1 activity. Atomt1 is a knockout mutant and the expression profile of the AtOMT1 gene has been determined as well as the consequences of the mutation on lignins, on soluble phenolics, on cell wall digestibility, and on the expression of the genes involved in monolignol biosynthesis. In this mutant and relative to the wild type, lignins lack syringyl (S) units and contain more 5 hydroxyguaiacyl units (5-OH-G), the precursors of S-units. The sinapoyl ester pool is modified with a two-fold reduction of sinapoyl-malate in the leaves and stems of mature plants as well as in seedlings. In addition, LC-MS analysis of the soluble phenolics extracted from the seedlings reveals the occurrence of unusual derivatives assigned to 5-OH-feruloyl malate and to 5-OH-feruloyl glucose. Therefore, AtOMT1 enzymatic activity appears to be involved not only in lignin formation but also in the biosynthesis of sinapate esters. In addition, a deregulation of other monolignol biosynthetic gene expression can be observed in the Atomt1 mutant. A poplar cDNA encoding a caffeic acid OMT (PtOMT1) was successfully used to complement the Atomt1 mutant and restored both the level of S units and of sinapate esters to the control level. However, the over-expression of PtOMT1 in wild-type Arabidopsis did not increase the S-lignin content, suggesting that OMT is not a limiting enzyme for S-unit biosynthesis. PMID- 12777057 TI - Serum anti-cholesterol antibodies in chronic hepatitis-C patients during IFN alpha-2b treatment. AB - Previously we detected more than 3 times higher anti-cholesterol antibody (ACHA) levels in HIV positive patients compared to healthy individuals, however, this level significantly decreased during highly active anti-retroviral therapy (HAART). In our present study we examined whether these findings could also be detected in patients with chronic hepatitis C (CHC). We calculated the correlation between the ACHA levels and the C5b-9 complement activation product. 39 patients with CHC were treated with IFN-alpha-2b (Schering-Plough) 5 MU daily for 6 weeks, followed by 5 MU TIW. Serum levels of ACHA and complement activation products were measured with ELISA. Serum HCV RNA was measured by a highly sensitive branched DNA technique before and 3, 6 and 12 months after the beginning of IFN-alpha-2b therapy. 52 healthy persons served as controls. At the onset of treatment ACHA level was significantly (p = 0.0062) higher in patients (40 (24-69) AU/ml) (median (interquartile range)) than in control sera (26 (20 35) AU/ml). In the 26 responder patients ACHA levels decreased to the normal level during the therapy, but no change was observed in the 13 non-responders. In patients with a sustained response ACHA levels remained low till the end of the 12 months IFN treatment. ACHA levels were significantly (p = 0.0422) higher in the patients with low (< 4.0 mmol/l) than in those with normal (> or = 4.0 mmol/l) cholesterol concentrations. The ACHA level before the therapy strongly correlated (r = 0.5499, p = 0.0014) with C5b-9 serum levels. ACHA levels are elevated in CHC, but this elevation is not as high as in HIV. Decrease of viral load by IFN-alpha-2b treatment in the responders results in normalization of ACHA concentration. High ACHA levels in patients with low serum cholesterol concentration suggest that high ACHA levels may contribute to the decrease in cholesterol levels. The correlation between the ACHA and C5b-9 levels indicate, that the ACHA may play a role in the complement activation in CHC. PMID- 12777058 TI - Anaphylactic and non-anaphylactic murine IgG1 differ in their ability to bind to mast cells: relevance of proper glycosylation of the molecule. AB - We have previously shown that murine IgG1 antibodies comprise two functionally distinct types regarding their ability to induce mast cell degranulation. In this work, we identified two IgG1-producing hybridomas, both with the same antigenic specificity (anti-DNP), but different in vivo anaphylactic activities. Whereas one of them secretes the anaphylactic IgG1 antibody, as assessed by passive cutaneous anaphylaxis, the other produces the non-anaphylactic IgG1 molecule. The evaluation of the ability of both types of IgG1 to bind to and activate a mouse mast cell line revealed that the anaphylactic IgG1 has a higher binding capacity and releases more beta-hexosaminidase from mast cells than the non-anaphylactic IgG1. Aglycosylated IgG1 obtained by treatment of the anaphylactic IgG1-producing hybridoma line with an inhibitor of N-glycosylation failed to elicit anaphylaxis. In addition, a goat anti-mouse IgG1 antibody reacted less with this aglycosylated IgG1 than with the glycosylated form. These results suggest that the anaphylactic activity of IgG1 antibodies is closely related to their structural conformation and the proper N-glycosylation of these molecules. Finally, the difference in the anaphylactic property between the two types of IgG1 seems to be primarily due to binding to the mast cell surface. PMID- 12777059 TI - Soluble cytokines can act as effective adjuvants in plasmid DNA vaccines targeting self tumor antigens. AB - There are few vaccination strategies available for the reproducible generation of a cytotoxic T cell (CTL) response, particularly in the setting of immunizing against a tumor antigen. Plasmid-based DNA vaccination offers several advantages as compared to MHC class I peptide-based vaccines or DNA immunization using viral vectors. Plasmid-based DNA vaccines are easily produced, can potentially elicit both an MHC class I and class II response, and have little infectious potential. Plasmid-based vaccines, however, have been poorly immunogenic. The systemic immune response generated after plasmid vaccination relies on in vivo transfection of local antigen presenting cells (APC) and both direct presentation and "cross priming" of antigen by professional and non-professional APC. Therefore, methods to enhance the function of APC, such as simultaneous inoculation with plasmids encoding cytokine genes, has resulted in an enhancement of detectable immunity after vaccination. We questioned whether local application of soluble cytokines would be effective in enhancing the systemic immune response elicited after DNA vaccination. Using a self-tumor antigen model, we vaccinated rats with a plasmid-based rat neu intracellular domain (ICD) DNA construct and either no adjuvant, soluble GM-CSF, or IL-12. We demonstrate that the addition of soluble GM-CSF or IL-12 to rat neu ICD DNA vaccination elicits detectable neu specific T cell immunity; specifically the generation of CTL. Antibodies directed against rat neu were not elicited with this approach, indicating that the neu specific T cell immune response elicited with plasmid DNA was skewed towards cell mediated rather than humoral immunity. PMID- 12777060 TI - Adenoviral-mediated delivery of a viral chemokine binding protein blocks CC chemokine activity in vitro and in vivo. AB - Chemokines are important mediators of leukocyte recruitment and activation that play critical roles in the pathology of inflammatory diseases such as atherosclerosis, rheumatoid arthritis and asthma. The vaccinia virus (strain Lister) expresses a 35 kDa soluble protein ('35K') that binds and inactivates a wide range of CC chemokines. We generated a recombinant adenovirus encoding soluble 35K (Ad35K). Ad35K-infected cell culture medium, containing recombinant 35K, potently reduced migration of CCR5-transfected 293 cells by 95% in response to the CC-chemokine RANTES, but had no effect on cells transfected with the CX3CR1 fractalkine receptor. Delivery of Ad35K to mice in vivo via tail vein injection resulted in expression of recombinant 35K in plasma and increased serum RANTES and MIP-1alpha levels when quantified by ELISA. However, chemotaxis of both CCR5-transfected cells and primary macrophages was inhibited by more than 90% by plasma from Ad35K-infected animals compared with control plasma from animals injected with AdGFP. Furthermore, 35K delivered by intra-peritoneal injection more than halved biogel-induced inflammatory cell recruitment in peritoneal exudates compared to AdGFP medium. These studies identify broad spectrum CC-chemokine blockade using in vivo adenoviral-mediated recombinant 35K expression as a promising strategy to reduce local and systemic inflammation. PMID- 12777061 TI - Reconstitution of the cellular immune response by lactoferrin in cyclophosphamide treated mice is correlated with renewal of T cell compartment. AB - Cyclophosphamide is an alkylating agent used to treat both malignant and non malignant immune-mediated inflammatory disorders in humans. It is also known as a potent immunosuppressive drug in humans and experimental animals. The aim of this study was to evaluate the effects of oral administration of lactoferrin (LF) on cellular responses and reconstruction of the lymphocyte pool in mice treated with cyclophosphamide (CP). Twelve week-old CBA mice were given a single intraperitoneal (i.p.) dose of CP (400 mg/kg body weight), then were treated per os with seven doses of LF (1 mg/dose) on alternate days. We demonstrated that the magnitude of delayed type hypersensitivity to ovalbumin, strongly diminished by CP action, was reconstituted by LF. Oral LF treatment also resulted in partial recovery of Concanavalin A-induced splenocyte proliferation. Blood profile analysis revealed elevation of leukocytosis by LF in CP-treated mice (from 64.9 to 84.76% of the control value). LF also caused substantial restoration of the percentage of the lymphocyte population in circulating blood (from 43.4 to 60.2% of the control values). LF alone had no effect on the neutrophil/lymphocyte ratio in normal mice, however, the total number of leukocytes decreased by 23.25%. Furthermore, we showed that LF increased the cellularity of spleens isolated from CP-treated mice (from 53.2 to 78.8%) and the content of peritoneal and alveolar macrophages (elevations from 50.6 to 67.3% and from 65.2 to 83.6%, respectively). Lastly, using panning technique, we demonstrated that LF strongly elevated the pool of CD3+ T cells in normal and CP-immunocompromised mice and CD4+ T cell content. In conclusion, we showed for the first time that lactoferrin, given orally to CP-immunosuppressed mice, could reconstitute a T-cell mediated immune response by renewal of the T cell pool. PMID- 12777062 TI - Influence of first-wave derived T lymphocytes in the long term functional reconstitution of allogeneic T cell deficient hosts. AB - The functional immunological reconstitution and the patterns of cytokine secretion were comparatively studied in BALB/c nu/nu mice grafted with allogeneic B6.Thy-1.1+ E14 or E18 embryonic thymus. In spite of equivalent proliferative responses to both mitogen or MLR stimuli, the two groups presented different cytokine patterns. B6 E18-thymus grafted BALB/c nu/nu mice showed a predominant IL-2/IFN-gamma secretion in response to mitogen or to CBA haplotype, with insignificant secretion of either cytokine to the tolerated BALB/c or donor B6 haplotype. In contrast, E14 grafted mice showed a significant IL-10 secretion, both in response to mitogens or to the tolerated haplotypes, even in the absence of a detectable proliferative response. A significant IFN-gamma secretion appeared only accompanying high responses to CBA. The preferential Th2 profile associated to the E14 chimeras was coincident with a longer lifespan of the nude host kept in a conventional environment, higher CD3+ cells frequency in the blood and functional restoration of allogeneic skin graft rejection, not seen on the E18 chimeras. The meaning of these results is discussed in relation to the previously described longer persistence of the first-wave donor derived lymphocytes in the allogeneic BALB/c periphery, also exclusive of the E14 grafted group. PMID- 12777063 TI - Localization of GPI-80, a beta2-integrin-associated glycosylphosphatidyl-inositol anchored protein, on strongly CD14-positive human monocytes. AB - Human monocyte/macrophage systems are extremely heterogeneous. Although many attempts have been made to define monocyte subpopulations, few antigens distinguish them. We previously reported that GPI-80, a novel glycosylphosphatidyl-inositol (GPI)-anchored protein that is expressed mainly on human neutrophils regulates neutrophil adherence and migration, and that GPI-80 is expressed on monocytes. In this study, we examined the precise distribution of GPI-80-positive monocytes using flow cytometry. Using anti-CD14 and anti-CD16 mAbs, almost all GPI-80-bearing monocytes belong to the strongly CD14-positive monocyte subpopulation. Furthermore, flow cytometric analysis of GPI-80 and other monocyte markers revealed that GPI-80 expression was high in CD11b-, CD32-, and CD64-positive monocytes. In contrast, GPI-80 expression was low in HLA-DQ positive monocytes. These results suggest that almost all GPI-80 positive monocytes belong to a monocyte subpopulation that is superior in phagocytosis and reactive oxygen production, but inferior in antigen presentation. GPI-80 may be a useful antigen for classifying monocytes into subpopulations. PMID- 12777064 TI - Synthetic mycoplasma-derived lipopeptide MALP-2 induces maturation and function of dendritic cells. AB - Dendritic cells (DC) modulate immune responses depending on the nature of the antigens. Receptors capable of discriminating these antigens on the basis of the pathogen-associated molecular patterns (PAMP) belong to the Toll-like receptor (TLR) family. The macrophage-activating lipopeptide 2 kDa (MALP-2), a synthetic lipopeptide derived from Mycoplasma fermentans, signals through TLR-2 and TLR-6. The aim of this study was to examine whether MALP-2 can modulate the functional properties of human monocyte-derived DC. The effects of this treatment were compared to those of the TLR-4 agonist lipopolysaccharide (LPS). To ensure clinical applicability, DC were generated under serum-free conditions. MALP-2 and LPS stimulation induced the expression of CD83 and increased the expressions of CD80, CD86, HLA-ABC and CD40. Furthermore, both substances decreased the endocytotic capacity of DC and induced the release of bioactive TNF-alpha and IL 10, whereas LPS additionally increased IL-12 release. Pretreatment with both substances boosted the allostimulatory capacity of DC. In a coculture with autologous lymphocytes, either MALP-2 or LPS pretreated DC induced a marked proliferation of lymphocytes, but only DC prestimulated with MALP-2 activated lymphocytes to produce the cytokines IL-4, IL-5 and IFN-gamma. No polarisation of lymphocytes into T-helper (Th)1 or Th2 was detected. These data indicate that MALP-2 is a potential candidate to modulate DC for clinical applications. PMID- 12777065 TI - Molecular diversity of oligotrophic and neurotropic members of the black yeast genus Exophiala, with accent on E. dermatitidis. AB - Analysis of ITS rDNA of the black yeast Exophiala dermatitidis revealed a close phylogenetic relationship to the meristematic fungus Sarcinomyces phaeomuriformis. As most strains of S. phaeomuriformis have a yeast-like phenotype corresponding to the anamorph genus Exophiala, a new combination in Exophiala is proposed. On the basis of ITS sequence, M-13 fingerprint and SSU intron data, two main entities could be distinguished within E. dermatitidis. One of these (B) contained prevalently strains from environmental sources, while the other (A) mainly comprised strains from clinical sources. This may be due to a difference in virulence. All strains from severe brain and disseminated infections in East Asia clustered in group A. However, strains of group A caused a relatively mild fungemia in patients outside East Asia. PMID- 12777066 TI - Bacterial host strains that support replication of somatic coliphages. AB - Somatic coliphages detected by Escherichia coli strain WG5 have been proposed as potential indicators of water quality. Their potential replication in the water environment is considered a drawback for their use as indicators. However, the contribution of replication outside the gut to the total numbers has never been quantified. It has not been determined either the fraction of bacterial strains that might support replication of phages detected by strain WG5 in the water environment. We examined the sensitivity of 291 host strains to 25 phages by streaking slants of the presumptive host strain onto an agar layer that contains bacteriophages, which gives a total of 7275 combinations (sensitivity tests). Only a 3.02% of the tests showed sensitivity. Additionally, six environmental strains were used as hosts to count phages in sewage and seawater. Phages isolated on these strains were used to infect strain WG5. The environmental strains detected 1 log10 fewer phages than strain WG5 in sewage and seawater. The fraction of phages that were detected by the six strains and that also infected strain WG5 ranged from < 0.07% to < 2.0% of the total amount of bacteriophages detected by strain WG5 in the same samples. Our results confirm that less than 3% of naturally occurring hosts support replication of phages infecting E. coli. We conclude that the contribution of replication to the number of somatic coliphages detected in the aquatic environment is negligible. PMID- 12777067 TI - Oxylipins and ascospore morphology in the ascomycetous yeast genus Dipodascus. AB - Immunofluorescence microscopy was used to assess members of the yeast genus Dipodascus for the presence of 3-hydroxy oxylipins. Fluorescence was associated with the aggregating ascospores in all species tested, thus suggesting the association of 3-hydroxy oxylipins with these cells, especially the surrounding slime sheaths. An ultrastructural study of the ascospores revealed sheaths with indentations, probably caused by the close packing of the ascospores to form clusters. In addition, an increase in the neutral and glycolipid fractions as well as a decrease in the phospholipid fraction during ascosporogenesis in D. ambrosiae was found. PMID- 12777068 TI - Pichia (Komagataella) pseudopastoris sp. nov., a new yeast species from Hungary. AB - Four strains of an unknown yeast species were isolated from rotten willow samples which were collected in Hungary. Although their phenotypic characteristics suggested that they were conspecific with Pichia pastoris, investigation of their small (18S) and large (26S) subunit rDNA revealed that they belonged to an undescribed yeast species. The description of the new yeast species, Pichia (Komagataella) pseudopastoris is given. PMID- 12777069 TI - Characterization of a non-pigment producing Monascus purpureus mutant strain. AB - A characterization of a non-pigment producing mutant Monascus purpureus M12 compared with its parental strain Monascus purpureus Went CBS 109.07 has been performed aiming to investigate the relation between pigment biosynthesis and other characteristics of these fungi. A comparison has been made of morphological features, some physiological properties and biochemical activities of both strains. The albino mutant exhibits an anamorph life cycle, high conidia forming capability, slower radial growth rate and temperature sensitivity. The assimilation capacity of both strains for mono-, disaccharides and some alcohols is in the same range (Yx/c 0.2 - 0.35), while the red strain has a higher fermentation capacity. In a selected albino mutant, the growth rate, metabolic activity and capacity for production of typical for Monascus fungi secondary metabolites were reduced considerably. Hydrolytic activity towards natural substrates expressed through glucoamylase and protease was approximately 10 fold lower in the non pigment producing strain (0.05 - 0.08 U/mg protein and 0.01 - 0.07 U/mg protein respectively) compared with the red one. Important qualitative differences between both strains was found in fatty acid composition and in the production of citrinin and monacolin. The mutant strain possessed C17, C20 and C22 fatty acids and did not produce citrinin. PMID- 12777071 TI - A new meristematic fungus, Pseudotaeniolina globosa. AB - A new species of Pseudotaeniolina, a genus of anamorphic, melanized fungi with meristematic development, is described. The species is compared to morphologically similar taxa among which are Trimmatostroma and Coniosporium. Its novelty is supported by SSU (small subunit) and ITS (internal transcribed spacer) rDNA sequence data. PMID- 12777070 TI - Comparison of three molecular methods for typing Aeromonas popoffii isolates. AB - Three typing methods, restriction fragment length polymorphism (RFLP) of the 16S 23S intergenic spacer region (ISR), PCR amplification of the enterobacterial repetitive intergenic consensus (ERIC) and of the repetitive extragenic palindromic units (REP), were evaluated for typing 26 isolates of Aeromonas popoffii from different geographical origins. When the methods were independently studied, ERIC showed the highest discriminatory power. When the methods were combined, the best combination of two methods was ERIC with REP since strains showed a tendency to cluster according to their geographical origin. However, this tendency was reinforced with the addition of ISR-RFLP. PMID- 12777072 TI - phoR1, a gene encoding a new histidine protein kinase Myxococcus xanthus. AB - A soil bacterium able to undergo multicellular development and a coordinated gliding in swarms, requires an accurate regulatory network of phosphorelay proteins. Inorganic phosphate is a limiting nutrient in soil and its importance in regulation is critical. As a step towards studying phosphate regulation and its influence in the developmental process in this bacterium, we screened a Myxococcus xanthus library for clones with phosphatase activity, and found four different ones. The deduced sequence of one of the cloned inserts is similar to that of the classic transmembrane histidine protein kinase of the sensor family of the two-component signal transduction systems with a high sequence similarity to the sensor kinase in the Pho regulon of Bacillus subtilis PhoR. This gene has been named phoR1 and its deduced amino acid sequence consists of 455 residues with a predicted molecular mass of 48.5 kDa. The M. xanthus PhoR1 deduced sequence contains all the characteristic histidine protein kinase motifs in the same order and with the same spacing. A hydropathy profile indicates two membrane spanning segments located at the extreme N-terminus, according to the putative sensor role of this domain. A gene-disrupted mutant is unable to produce normal mature fruiting bodies and produces fewer spores. PMID- 12777073 TI - Influence of VEGF-R2 inhibition on MMP secretion and motility of microvascular human cerebral endothelial cells (HCEC). AB - Neovascularization and invasion are key features of malignant gliomas. Matrix metalloproteinases (MMPs) are supposed to play a major role mediating these processes. To analyze the expression patterns of MMPs in microvascular human cerebral endothelial cells (HCEC), we isolated endothelial cells from normal human brain microvessels. Characterization of cellular origin was performed by immunostaining, using the endothelial cell markers Ulex europaeus Agglutinin-1, von-Willebrand-Factor and Glucose-transporter-1. Contamination by other cell types was tracked by immunohistochemistry for GFAP (astrocytes), ASM (pericytes) and CD68 (macrophages). Secretion of MMPs was evaluated by ELISA and zymography. To determine whether HCEC show any difference in MMP expression compared to endothelial cells of other origin we analyzed human umbilical vein endothelial cells (HUVEC). HCEC show a decrease of MMP-3 and MMP-2 protein when treated with SU5416, a VEGF-R2 (KDR/flk-1) inhibitor, whereas MMP expression remained unchanged in HUVEC. To determine whether these findings show any effect in the motility of these cells we used a three-dimensional co-culture assay of avascular glioblastoma spheroids with primary HCEC spheroids. Untreated controls showed invasion of both cell populations into each other whereas treatment of the co cultures with SU5416 resulted in complete inhibition of endothelial cell invasion hence indicating that flk-1 related motility of endothelial cells is critically involved in this process and can be studied with this assay. The results of different effects of anti-angiogenic treatment on proteolytic properties of two endothelial cell populations suggest that neovascularization of human brain tumors in vitro is dependent on the surrounding endothelial cell type and should therefore be studied with organ-specific human microvascular cerebral endothelial cells. PMID- 12777074 TI - Secretory meningiomas. AB - Secretory meningiomas are a rare meningioma subtype. Among meningiomas, the frequency of secretory meningiomas is 1.6%. Unlike other meningioma types, most of the patients were female (ratio 3:1). No recurrence was reported during the 24 180 months follow-up period of our secretory meningiomas in which, a low level of 0.3% Ki-67 proliferative index was reported. In this meningioma subtype, the percentage of cases with positive progesterone receptor is 33%. With carcinoembryonic antigen, cytokeratin and epithelial membrane antigen, in all the cases positivity was observed in both, the inclusions and the cells surrounding them. With human milk fat globulin 2, a high ratio (92%) of positivity was observed. Majority of the cases were negative with CA125, only three of the cases had suspicious positivity. Distribution of inclusions was irregular and their positive reactions showed varying staining features. Positivity with alpha-1 antitripsin was seen not only in the inclusions but also in some meningothelial cells as well. Ubiquitin was positive in inclusions of the 83% of cases. Staining features of the inclusions pointed out the possibility of them being in a varying age and/or content. Secretory meningiomas are a different type compared to other meningiomas, not only with their histological features but also with their clinical features as well. PMID- 12777075 TI - Repetitive 5-aminolevulinic acid-mediated photodynamic therapy on human glioma spheroids. AB - The response of human glioma spheroids to repetitive 5-aminolevulinic acid mediated photodynamic therapy (PDT) was investigated. In all cases, light fluences were kept below toxic thresholds to simulate conditions typically found at 1-2 cm depths in brain adjacent to tumor. Significant inhibition of spheroid growth was observed following multiple PDT treatments at sub-threshold light fluences. The effect appears to be insensitive to the treatment intervals investigated (weekly or bi-monthly). In all cases, suppression of growth was observed for the duration of treatment. Low fluence rates (< or = 5 mW cm(-2)) appear to be more effective than high fluence rates (25 mW cm(-2)). No evidence of PDT resistance was found in this investigation. PMID- 12777076 TI - Formation of DNA adducts and tumor growth delay following intratumoral administration of DTI-015. AB - Intratumoral (IT) administration of DTI-015 (BCNU in 100% ethanol) utilizes solvent facilitated perfusion for the treatment of tumors. RIF-1 tumors were treated by IT injection of either ethanol alone or 0.05-1.0 mg of DTI-015 or by i.v. injection of 0.5 mg of BCNU. Treatment with ethanol alone or i.v. injection of 0.5 mg of BCNU did not produce a significant growth delay. In contrast, IT administration of DTI-015 produced a significant growth delay at each of the treatment doses (p < 0.05 to p < 0.001). We have quantified the levels of N7-(2 hydroxyethyl) guanine (N7-HOEtG) in RIF-1 tumors 24h following either IT treatment with 0.5 mg DTI-015 or i.p. administration of 0.5 mg BCNU. Levels of N7 HOEtG (micromol/mol DNA) were < or = 0.08 for both untreated controls and following i.p. treatment with BCNU and 13.1 +/- 5.6 following IT administration of DTI-015. The levels of N7-HOEtG detected in RIF-1 tumors following IT administration of DTI-015 were 164-fold higher than the level(s) of N7-HOEtG in the i.p. BCNU treated tumor samples. These studies demonstrate that IT administration of DTI-015 produces high levels of DNA adducts in the tumor which correspond to a significant increase in tumor growth delay compared to the same dose of BCNU administered systemically. PMID- 12777077 TI - Magnetic resonance imaging of patched heterozygous and xenografted mouse brain tumors. AB - Experimental mouse models are emerging as useful systems for the study of human brain tumors. Nuclear magnetic resonance imaging (MRI) methods can noninvasively provide images of complex heterogeneous tissues such as experimental brain tumors. The current report demonstrates the feasibility of longitudinal high resolution MRI in two mouse brain tumor models: patched heterozygous (ptc +/-) mice with spontaneously arising posterior fossa tumors that resemble human medulloblastoma, and homozygous nude mice implanted with intracerebral xenografts of human medulloblastoma cell lines. Methods were optimized to achieve favorable volumetric comparison with histologic methods and sub-millimeter resolution, improved by contrast enhancement with intravenous administration of a gadolinium based agent. Results also show that experimental mice, even symptomatic mice, tolerate repeated serial imaging studies over weeks to months to follow tumor progression and to visualize placement of an intracerebral drug delivery system. PMID- 12777078 TI - Rapid shrinkage of remnant central neurocytoma after gamma knife radiosurgery: a case report. AB - Central neurocytomas are typically benign tumors that have high local control rates after gross total resection. Radiotherapy for residual or recurrent central neurocytomas is controversial. We report a 30-year-old male with a central neurocytoma in the lateral ventricle. The tumor was subtotally resected through a transcallosal approach, and subsequently treated with gamma knife radiosurgery. The tumor had shrunken markedly by 2 months after radiosurgery and remained unchanged during the one year follow-up period. Gamma knife radiosurgery may be an option for effective treatment of patients with residual or recurrent central neurocytomas. PMID- 12777079 TI - Metaplastic bone formation in a low grade conus glioma: case report and review of the literature. AB - Calcifications associated with both benign and neoplastic intra-axial lesions of the central nervous system (CNS) are well recognized. Bony metaplasia in the CNS, where there is formation of mature trabecular bone, is a much more rare entity and has not been reported in the spinal cord. We present a case of bony metaplasia in the conus medullaris associated with a low grade astrocytoma. Radiological, pathological and clinical features of this unique case are discussed. PMID- 12777080 TI - Successful high-dose chemotherapy for widespread neuroaxis dissemination of an optico-hypothalamic juvenile pilocytic astrocytoma in an infant: a case report. AB - We report a 13-month-old boy with diencephalic syndrome (DS) due to an optico hypothalamic juvenile pilocytic astrocytoma (JPA). Massive neuroaxis dissemination was identified at diagnosis. He received 6 courses of combined conventional-dose chemotherapy consisting of carboplatin (CBDCA), etoposide (VP 16), and cyclophosphamide (CPA) followed by high-dose chemotherapy with CBDCA, CPA, and ranimustine (MCNU) and peripheral blood stem cell transplantation (PBSCT). This treatment produced tumor regression in both intracranial and spinal lesions and remarkable improvement of DS. The rare combination of DS and symptomatic neuroaxis dissemination of JPA at diagnosis suggests that the behavior of some of these tumors is more aggressive and resistant to conventional dose chemotherapy than is that of JPA without DS manifestation and dissemination. PMID- 12777081 TI - Postoperative management of patients after stereotactic biopsy: results of a survey of the AANS/CNS section on tumors and a single institution study. AB - As little consensus exists on the postoperative care of patients undergoing stereotactic biopsy, we sought to establish a new algorithm for their postoperative management. First, we surveyed active members of the AANS/CNS Section on Tumors to determine national practice patterns for patients after stereotactic biopsy. Second, we retrospectively reviewed 84 consecutive stereotactic biopsy procedures at our institution to assess the potential benefit of routine computed tomography (CT) scanning and intensive care unit (ICU) monitoring. Finally, we prospectively applied this new algorithm in 54 patients to assess its validity. Of 629 surgeons, 263 (42%) responded; they were experienced neurosurgeons (mean 15 years in practice) who performed more than 10 stereotactic biopsies per year. Most surgeons (59%) routinely ordered postoperative CT scans, and the remainder ordered scans based on specific indications. Patients were transferred from the recovery room to a special care unit (47%), regular room (47%), or home (6%). In our retrospective review, 81 patients underwent 84 stereotactic biopsy procedures; 79 underwent postoperative CT scanning and all 81 were monitored overnight in the ICU. Among five (6%) patients who experienced intraoperative hemorrhage, two (2%) underwent craniotomy to control arterial bleeding. Three (4%) patients developed new neurological deficits, which occurred within 2 h of surgery. In both groups, CT scans were helpful in excluding hemorrhage that would require re-operation. In the remaining patients (90%), findings on routine postoperative CT did not alter patient management and ICU monitoring appeared unnecessary because neurological complications occurred within 2 h postoperatively. We confirmed these results in the prospective study of 54 patients undergoing stereotactic biopsy without routine postoperative CT scanning or ICU monitoring. In contrast with national practice patterns reported, we recommend that CT scanning and ICU monitoring be reserved for patients who have intraoperative hemorrhage or new deficits after surgery. All other patients can be monitored for 2 h in the recovery room and transferred to a regular hospital room without a postoperative CT scan. PMID- 12777082 TI - Intracavitary VEGF, bFGF, IL-8, IL-12 levels in primary and recurrent malignant glioma. AB - Intracavitary levels of VEGF, bFGF, IL-8 and IL- 12 were evaluated by ELISA in 45 patients, 7 with recurrent anaplastic astrocytoma (rAA), 12 with glioblastoma (GBM) and 26 with recurrent glioblastoma (rGBM). In 25 patients plasma levels of the molecules were also quantitated. Twenty-three healthy controls were also studied for plasma concentrations of the same molecules. Plasma levels of VEGF (mean 33.89 +/- 6.71 pg/ml) and bFGF (mean 11.1 +/- 3.24 pg/ml) were higher in patients than in controls (mean 16.78 +/- 3.7 pg/ml for VEGF, mean 0.21 +/- 0.09 pg/ml for bFGF) (p = 0.04 and p = 0.001, respectively) while plasma IL-12 levels were lower (mean 45.6 +/- 1.5 pg/ml in patients, mean 79.7 +/- 1.3 pg/ml in controls) (p = 0.009). Intracavitary VEGF levels were 5-53.307 fold higher (mean 90,900 +/- 24,789 pg/ml) than in the corresponding plasma. Also IL-8 concentrations were higher in intracavitary fluid (mean 6,349.76 +/- 1,460.93 pg/ml) than in plasma (mean 43.44 +/- 24.82 pg/ml). Maximum VEGF levels were found in tumor fluid of recurrent glioblastoma patients (mean 147,678 +/- 39.903 pg/ml), intermediate levels in glioblastoma patients (mean 20,322 +/- 11,892 pg/ml) and lower levels in rAA patients (mean 9,111 +/- 5,789 pg/ml). The data also suggest that higher intracavitary levels of VEGF and IL-8, and lower IL-12 levels, may be correlated with shorter adjunctive survival times, but more data will need to be collected to establish this correlation clearly. PMID- 12777083 TI - Cerebral glucose metabolism in long-term survivors of childhood primary brain tumors treated with surgery and radiotherapy. AB - Delayed structural cerebral sequelae has been reported following cranial radiation therapy (CRT) to children with primary brain tumors, but little is known about potential functional changes. Twenty-four patients were included, diagnosed and treated at a median age of 11 years, and examined after a median recurrence free survival of 16 years by MRI and Positron Emission Tomography using the glucose analog 2-18F-fluoro-2-deoxy-D-glucose (18FDG). Three patients were not analyzed further due to diffuse cerebral atrophy, which might be related to previous hydrocephalus. Twenty-one patients were evaluable and regional cerebral metabolic rate for glucose (rCMRglc) was estimated in nontumoral brain regions in 12 patients treated with surgery alone and 9 patients treated with both surgery and CRT. Furthermore 10 normal controls matched for age at examination were included. Patients treated with both surgery and CRT had a general decreased rCMRglc compared to normal controls and patients treated with surgery alone, significantly (p < 0.05) in 5 of 11 regions of interest. No difference was found in rCMRglc between normal controls and patients treated with surgery alone. We conclude that there is a general reduction in rCMRglc in long term recurrence free survivors of childhood primary brain tumors treated with CRT in high doses (44-56 Gy). PMID- 12777084 TI - A case of recurrent subependymoma with subependymal seeding: case report. AB - Subependymoma is a rare, slow growing, rarely recurrent tumor. We report a case of re current subependymoma with subependymal seeding. An intraventricular tumor in the left temporal horn was detected in a 48-year-old female who presented with a 4-year history of dizziness and memory disturbance. Following near total surgical resection, a tumor diagnosis of subependymoma was confirmed by scattered clusters of isomorphic nuclei embedded in a dense fibrillary matrix of glial cell processes. Twenty-six months after surgery, follow-up (F/U) magnetic resonance (MR) imaging revealed tumor recurrence in the previous site which necessitated linear accelerator radiosurgery (LINAC). A further 21 months later, F/U MR imaging showed recurrent, multiple, enhanced, nodular lesions in the enlarged left lateral ventricle for which the patient underwent reoperation. Radiological and operative findings revealed local relapse with subependymal seeding. The pathological finding was similar to that of the previous tumor and compatible with subependymoma. The patient underwent radiation therapy for the residual tumor. This case history suggests that symptomatic residual tumors require close observation even though the clinical course of subependymoma is usually benign. PMID- 12777085 TI - Locoregional radioimmunotherapy in selected patients with malignant glioma: experiences, side effects and survival times. AB - Prognosis of malignant glioma is very unfavourable mainly due to minimal tumour remnants in the peritumoural tissue. Intralesionally applied radioimmunotherapy is a possible therapeutical option with the potential to improve survival of patients with malignant glioma. We investigated side effects and survival after surgery, conventional radiotherapy and additional radioimmunotherapy with labelled tenascin-antibodies in patients with malignant glioma. METHODS: Since 1995, 37 patients were treated with radioimmunotherapy after resection and radiotherapy of a malignant glioma. Patients received antibodies labelled with yttrium-90 and iodine-131 in different doses into the tumour cavity via a previously implanted ommaya-reservoir. Treatment was applied in up to 8 cycles (mean 2.96 cycles) in time intervals of 6-8 weeks. Mean age was 46 years, histology was anaplastic astrocytoma in 13 patients and glioblastoma in 24 patients. RESULTS: For the whole group median survival time has not yet been reached. For glioblastoma the median survival time is 17 months, 5-year survival probability for anaplastic astrocytoma is 85% approximately. Quality of life was acceptable. Acute side effects following treatment were headache, seizures and worsening of pre-existing neurological symptoms. Late side effects were skin necrosis and, in 1 case, a delayed aphasia probably due to a vascular lesion. CONCLUSION: Radioimmunotherapy prolonged survival time in a selected group of patients with malignant gliomas as compared to a historical control group. Patients with anaplastic astrocytomas seem to have more benefit from this therapy than patients with glioblastomas. PMID- 12777086 TI - Evaluation of brain tumor metabolism with [11C]choline PET and 1H-MRS. AB - BACKGROUND: The signal of choline containing compounds (Cho) in proton magnetic resonance spectroscopy (1H-MRS) is elevated in brain tumors. [11C]choline uptake as assessed using positron emission tomography (PET) has also been suggested to be higher in brain tumors than in the normal brain. We examined whether quantitative analysis of choline accumulation and content using these two novel techniques would be helpful in non-invasive, preoperative evaluation of suspected brain tumors and tumor malignancy grade. METHODS: 12 patients with suspected brain tumor were studied using [11C]choline PET, gadolinium enhanced 3-D magnetic resonance imaging and 1H-MRS prior to diagnostic biopsy or resection. Eleven normal subjects served as control subjects for 1H-MRS. RESULTS: The concentrations of Cho and myoinositol (mI) were higher and the concentration of N acetyl signal/group (NA) lower in brain tumors than in the corresponding regions of the normal brain. There were no significant differences in metabolite concentrations between low- and high-grade gliomas. In non-tumorous lesions Cho concentrations were lower and NA concentrations higher than in any of the gliomas. Enormously increased lipid peak differentiated lymphomas from all other lesions. The uptake of [11C]choline at PET did not differ between low- and high grade gliomas. The association between Cho concentration determined in 1H-MRS and [11C]choline uptake measured with PET was not significant. CONCLUSION: Both 1H MRS and [11C]choline PET can be used to estimate proliferative activity of human brain tumors. These methods seem to be helpful in differential diagnosis between lymphomas, non-tumorous lesions and gliomas but are not superior to histopathological methods in estimation of tumor malignancy grade. PMID- 12777087 TI - Primary medulla oblongata germinoma: a case report and review of the literature. AB - We describe a case of primary intracranial medulla oblongata germinoma in a 16 year-old girl who presented with progressive headache and blurred vision. Magnetic resonance imaging demonstrated a heterogeneous exophytic mass arising from dorsal aspect of medulla oblongata with extension into fourth ventricle. The differential diagnosis for this patient had included ependymoma, exophytic glioma, medulloblastoma and choroid plexus papilloma. After surgical resection and radiation therapy, she remains alive and recurrence-free for 7 years. PMID- 12777088 TI - Radiotherapy to primary CNS germinoma: how large an irradiated volume is justified for tumor control? AB - Between 1990 and 1999, there were 30 primary central nervous system (CNS) germinoma patients who received radiotherapy (RT) as treatment. Of these, 23 are male and 7 are female patients, with a median age of 16 years. The treatment field of RT included whole neuraxis in 10, whole brain in 8 and local tumor site in 12 patients; the median dose delivered to the whole neuraxis being 3060 cGy, with 3060 cGy to the whole brain and 5040 cGy to the tumor site. Chemotherapy was prescribed in 9 patients. The median time on follow-up for survivors is 73 months. There were 7, out of a total of 30 patients, who suffered treatment failure. Five of twelve patients (41.6%) who received partial brain RT failed in the brain, with no difference in the rate between patients with or without chemotherapy, and only 2 of 18 patients (11.1%) who received whole brain or whole neuraxis RT failed in the brain (p = 0.053). None of 5 spinal seeding patients failed in the spine and only one failed in the brain after whole neuraxis RT, one patient without whole neuraxis RT (5%) failed in the spine. In summary, partial brain RT will have higher probability of intracranial relapse, and sparing the spinal RT will not result in more spinal failure, whole brain RT would be sufficient for tumor control on primary CNS germinoma. PMID- 12777089 TI - Paraneoplastic antineuronal antibodies in patients with systemic autoimmune diseases. AB - Sera from 71 patients with primary Sjogren's syndrome (PSS) and from 102 patients with systemic lupus erythematosus (SLE) were tested by immuno-dot blotting against HuD, Ri, Yo and amphiphysin recombinant proteins. For Ri, Yo and amphiphysin antigens, no immunoreactivity was found in the 173 sera tested. One PSS patient with a clinical picture of subacute sensory neuronopathy had high titers of anti-Hu antibodies. An extensive search for an underlying tumor was initially negative but a small cell lung cancer was eventually discovered three years later. Another patient with SLE and a clinical picture of demyelinating polyradiculoneuropathy had anti-Hu antibodies. Repeated search for an underlying tumor remains negative after five years follow-up in this young non-smoking patient. In addition, the neuropathy progressively improved and the anti-Hu antibodies titer slowly decreased from 1:8000 to 1:2000, making the diagnosis of paraneoplastic syndrome unlikely in this patient. This study indicates that the detection of anti-Hu antibodies in patients with known symptomatic systemic autoimmune diseases such as PSS or SLE should induce the same work-up than the detection of these antibodies in the absence of other immune diseases, i.e. repeated search for occult cancer during several years. As illustrated by our first patient, this strategy may be fruitful. Nevertheless, the clinician should know that anti-Hu antibodies may exceptionally (0.6% in this series) occur in systemic autoimmune disorders with neurological complications, in the absence of an underlying neoplastic disease. PMID- 12777090 TI - Nervous system lymphoid infiltration in Waldenstrom's macroglobulinemia. A case report. AB - Neurological complications may occur in the course of Waldenstrom's macroglobulinemia (Wm). These complications affect both the central and peripheral nervous system. Pathogenesis is debated and several mechanisms have been suggested. Direct malignant lymphoid infiltration (MLI) is very rare and has only been reported in a few cases. Case report. A 67-year-old woman suffered from Wm for 10 years before she presented neurological complications of the disease. She did not receive any specific treatment for Wm owing to a lack of symptoms. The neurological complications consisted of peripheral demyelinating polyneuropathy and MLI of the nervous system. This infiltration was located in the lower perimedullary and cauda equina roots regions, with possible centromedullary involvement. Indirect demonstration of MLI was achieved through non-invasive tests including magnetic resonance imaging and detailed cerebrospinal fluid examination. General and intrathecal chemotherapy led to clinical, biological and radiological improvement over a four-year period. PMID- 12777091 TI - Dysembryoplastic neuroepithelial tumor or dysembryoplastic cortical neurocytoma? PMID- 12777092 TI - Construct models in veterinary behavioural medicine: lessons from the human experience. AB - Veterinary behavioural medicine is marked by inconsistency in the naming and description of animal behavioural disorders, potentially hindering research and reducing the clinical utility of diagnoses. Sources of diagnostic inconsistency are suggested and parallels with human psychiatry are identified. This paper questions the use of categorical models as the basis for classification and reviews criticisms of categorical approaches in psychiatry. Evidence is presented to suggest an inconsistency between discrete categories and the structure of animal behavioural disorders. The potential benefits of exploring alternative models are discussed. It is concluded that it is important to move away from an approach based on advocacy and towards objective assessment of all available data. Existing diagnoses should be viewed as hypothetical constructs, with the underlying hypotheses being identified and subject to investigation. It is emphasized that global communication, empirical investigations and critical review are necessary for the development of a solid scientific basis within veterinary behavioural medicine. PMID- 12777094 TI - Early weaning as cause of malnutrition in half-bred zebu calves. PMID- 12777093 TI - The effect of subclinical hypocalcaemia induced by Na2EDTA on the feed intake and chewing activity of dairy cows. AB - The effects of induced subclinical hypocalcaemia (SCHC) on feed intake and chewing activity during eating and rumination were studied in dairy cows. Two non lactating and non-pregnant cows were subjected to three different treatments, with one test per day, such that the plasma free (ionized) calcium (FCa) concentration was maintained at the eucalcaemic level or at one of two constant SCHC levels. The cows and test days followed a 2 x 3 crossover design. SCHC was maintained for 7 h by repeatedly infusing 5% Na2EDTA so that constant FCa concentrations of 0.8 mmol/L or 0.6 mmol/L in plasma were achieved. Control conditions were achieved by infusing isotonic saline. Feed intake and the number of the rumination periods were recorded during test days. The proportion of feed eaten during each test meal (EatPro) was related to the mean plasma FCa. An almost linear decrease in EatPro was observed when the plasma FCa was 0.6-0.9 mmol/L. The cows showed no other clinical signs of hypocalcaemia during Na2EDTA infusion. The time spent chewing during eating and rumination, and the number of rumination periods during a test day, decreased with a decline in plasma FCa concentration. It was concluded that induced SCHC depresses the feed intake and ruminative activity of dairy cows. PMID- 12777095 TI - Species differences in the susceptibility of erythrocytes exposed to free radicals in vitro. AB - The susceptibility of human, cow, pig, sheep and rabbit erythrocytes to free radicals (peroxyl radicals) generated in vitro by 2,2'-azo-bis(2-amidinopropane) hydrochloride (AAPH) was evaluated by means of a haemolysis test and expressed as the time to 50% of maximal haemolysis (HT50). The most sensitive to damage by free radicals appeared to be the erythrocytes of pigs and sheep, their HT50 values being (mean +/- SEM) 85.1 +/- 1.28 min and 89.0 +/- 1.31 min, respectively. Human erythrocytes and those of cows and rabbits were about twice as resistant, their HT50 values being (mean +/- SEM) 174.3 +/- 1.53 min, 181.2 +/ 1.22 min and 183.4 +/- 2.54 min, respectively. Pig and sheep erythrocytes used in the haemolysis test provided an indication of the antioxidant status in a shorter time (2.5 h versus 4.5 h) than those of the other species studied. The results indicate that the HT50 test may be a convenient alternative to the osmotic resistance test for defining the antioxidant resistance of erythrocytes. PMID- 12777096 TI - The effects of breed, neonatal age and pregnancy on the plasma copper status of goats in Oman. AB - There is a history of copper deficiency in grazing Omani livestock and the copper status of three economically important goat breeds, Jabal Akhdar (JA), Batina (B) and Dhofari (D) were therefore compared in October/November (cool season) and June (dry season) in a penned flock given a plentiful dietary supply of copper. In the cool season, 62 lactating does (5 JA, 12 B and 33 D), their 0-5-day-old kids (17 JA, 19 B and 22 D) and 25 dry does (17 JA, 5 B and 3 D) were blood sampled. In the dry season, the does sampled were either barren (15 JA, 16 B and 13 D) or pregnant (9 JA, 13 B and 33 D). The samples were analysed for total copper (TCu) and trichloroacetic acid (TCA)-soluble copper (TCA-sol Cu). There were no effects of breed on TCu or TCA-sol Cu in the cool season, the overall means being 0.75 (SE 0.049) and 0.59 (SE 0.052) mg/L. The mean TCu was low in kids at birth (0.59 mg/L) but had increased to 0.86 mg/L by 4 days of age (p < 0.001); breed differences were found (p < 0.002), the pooled values for JA, B and D being 0.77, 0.59 and 0.68 (SE 0.033-0.044) mg/L, respectively. By the dry season, the mean TCu had risen in barren does to 0.96 (0.045) mg/L but not in pregnant does (0.76 (0.047) mg/L: p < 0.002) and breed differences had emerged, the mean for D being 20% lower than those for JA and B (p < 0.05). The highest TCu values were found in the 7-month-old kids in June ( 1.17 (0.039) mg/L) but the breeds did not then differ. Some effects on TCA solubility were found but were considered unreliable. Breed effects may have been diminished by the generous supply of copper and early stage of lactation studied. PMID- 12777097 TI - The early effects of Clostridium perfringens type D epsilon toxin in ligated intestinal loops of goats and sheep. AB - Clostridium perfringens type D produces enterotoxaemia in goats, sheep and other animals. The disease is caused by C. perfringens epsilon toxin and, while enterotoxaemia in goats is usually characterized by enterocolitis, the disease in sheep is characterized by systemic lesions (such as lung and brain oedema) with minor and inconsistent changes observed in the intestine. A possible explanation for these differences is that epsilon toxin is more promptly absorbed by the ovine than by the caprine intestine. In an attempt to clarify this, we examined the early effects of epsilon toxin on caprine and ovine intestine. Intestinal loop assays were performed to analyse the physiological and morphological changes induced by epsilon toxin in the intestine of these species. Fluid accumulation was observed in caprine and ovine ileum and colon treated with epsilon toxin. Ileal loops from goats treated with epsilon toxin retained sodium and water earlier than ovine ileal loops treated with the same toxin. Histological analysis showed morphological alterations in the colon of both species as early as 2 h after the commencement of epsilon toxin treatment: these changes were more marked in goats than in sheep. No morphological changes were observed in the ileum of either species after 4 h incubation with epsilon toxin. These results suggest that epsilon toxin modifies ion and water transport in the small and the large intestine of goats and sheep through different mechanisms. PMID- 12777099 TI - Microvasculature of the human cerebral white matter: arteries of the deep white matter. AB - The vascular architecture of the human cerebral deep white matter was studied using soft X-ray and diaphanized specimens, achieved by intra-arterial injection of barium and vascular stain respectively, and also by electron microscopic examination of the corrosion cast of arteries in normal adult brains. The deep white matter arteries passed through the cerebral cortex with a few branches to the cortex and ran straight through the white matter. The arteries concentrated ventriculopetally to the white matter around the lateral ventricle. Anastomoses were noted around the ventricular wall at the terminals of the deep white matter arteries. No centrifugal branches irrigating the periventricular white matter from the lenticulo-striate arteries were observed in the present study. The presence of anastomoses among the terminal branches of deep white matter arteries protects against ischemic change or infarction in this area from an occlusion of a single deep white matter artery. This may lead to development of terminal zone infarction from ischemia or vascular diseases, affecting multiple deep white matter arteries. The subcortical and deep white matter arteries had thick adventitial sheaths and large adventitial spaces in the white matter but not in the cortex. The presence or absence of the adventitial space is regarded as another characteristic difference between the arteries in the white matter and cortex. This difference may influence pathological changes in vascular lesions in these respective areas. PMID- 12777098 TI - Construction, expression, purification, refold and activity assay of a specific scFv fragment against foot and mouth disease virus. AB - An active form of a single-chain antibody (scFv) from the murine monoclonal antibody (mAb) 1C7, which is specific for type O foot and mouth disease virus (FMDV), was produced in Escherichia coli. The complementary DNAs encoding the variable regions of the heavy chain (VH) and light chain (VL) were connected by a (Gly4Ser)3 linker, using an assembly polymerase chain reaction. VH-(Gly4Ser)3-VL genes were screened by phage display technology. The sequencing results showed that the VH gene of scFv was composed of germline VH76-1BG-DFL16.1-JH4 and the VL gene of scFv consisted of germline bw20-JK2. The resultant scFv gene was cloned to the pPRoEX HTc vector and expressed in E. coli as inclusion bodies. After extraction from the E. coli cells, the inclusion bodies were solubilized and denatured in the presence of 8 mol/L urea. The expressed scFv fusion proteins were purified by nickel-nitrilotriacetic acid and finally renatured by dialysis. The purity and activity of the purified scFv were confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and enzyme-linked immunosorbent assay. The result revealed that the 1C7 scFv conserved the same characteristics of specific recognition and binding to type O FMDV as the parental 1C7 mAb. PMID- 12777101 TI - Microvasculature of the human cerebral meninges. AB - In the present study, the human cerebral meninges were rich in blood vessels, but no capillaries were noted. The meningeal arteries ran over the veins where they crossed. Several arterial anastomoses existed on the cortical surface. The meningeal arteries were classified into four parts; the conducting artery approximately 700 microm in diameter, distributing artery approximately 200 microm in diameter, precortical artery approximately 60 microm in diameter and cortical artery approximately 30-40 microm in diameter. A single distributing artery supplied the area of approximately 3.5 x 2.0 mm on the brain surface. They further ramified into precortical arteries which stemmed cortical arteries. These precortical arteries had the distributing area of 1 mm2 and this distributing area was the same size as the width of human ocular dominant column of the visual cortex. Constriction, like a sphincter, was observed at the bifurcation of the distributing arteries. The cerebral blood vessels, which regulated the blood flow and reacted to autonomic nerve stimuli, seemed to correspond to the distributing arteries. PMID- 12777100 TI - Immunohistochemical localization of human pineal tissue antigens in normal retina and retinoblastomas. AB - The normal human retina and retinoblastomas were examined immunohistochemically to assess the localization of pineal antigens in the retina and the oncogenesis and differentiation of retinoblastoma. In the present study, 41 eyes excised from children (aged 4 months to 7 years, all unilateral occurrence) diagnosed to have retinoblastoma and 4 eyes with normal retinas, were used. Retinoblastomas were histopathologically classified into well-differentiated, moderately differentiated, and poorly-differentiated types. The antibodies used were 9 monoclonal antibodies to human pineal antigens and 6 antibodies to neural tissues. In the normal retina, staining patterns characteristic of retinal cell layers were observed with PP1, PP3, PP5, PP6, PI1, and PI2 antibodies. In retinoblastomas, PP5 antibody, which reacts with horizontal cells and ganglion cells, and PP6 antibody, which reacts with part of the bipolar cells in the inner nuclear layer, showed intense staining in well-differentiated retinoblastomas, but the intensity of staining and the positivity decreased with the degree of dedifferentiation. Antigens recognized by PP3 and PP4 antibodies were positive in all retinoblanstomas. Reactions to GFAP antibody and antibodies that recognize Muller cells were negative. Retinoblanstomas may express markers of not only photoreceptor cells but also other retinal nuclear cells. These results suggest that the retinoblastoma might be developed from visual stem cells, which are common progenitor cells of photoreceptor cells, intermediate neurons, and ganglion cells. PMID- 12777102 TI - Creutzfeldt-Jakob disease with florid plaques after cadaveric dura mater graft. AB - A patient with dura-associated Creutzfeldt-Jakob disease (D-CJD) which occurred about 15 years after a dura mater graft is reported in the present study. The prion protein gene analysis disclosed no mutation. The D-CJD was atypical in: (i), the long interval between the onset of ataxia and the occurrence of dementia; (ii), the presence of transient myoclonus; and (iii), the presence of florid plaques in the brain. The electron-microscopic findings showed bundles of amyloid filaments which radiated from the center of the plaques without degenerating neurites. This case of D-CJD may belong to a new subtype of D-CJD. PMID- 12777104 TI - Glioblastoma with lymph node metastases. AB - Metastatic spread of malignant astrocytomas is rare and documented in very few patients with this tumor. Both pathologists and clinicians may confront more of these cases as the patients live longer. We present a 40-year-old-man with glioblastoma multiforme metastasizing to the supraclavicular lymph node after surgery. The tumor was located at the parietal convexity and several lymphadenopathies occured after surgery. PMID- 12777103 TI - Meningioangiomatosis with predominantly cellular pattern. AB - A case of meningioangiomatosis (MA), in a 10-year-old-girl with refractory complex partial and secondary generalized seizures, starting at the age of 8 years, is presented. MRI evaluation revealed a lesion located at the left frontal lobe; the patient underwent surgical lesionectomy. Histology revealed the lesion to have the features of MA. The patient is symptom-free a year postoperation. We report the histological, immunohistochemical and imaging findings in view of previous pertinent reports. PMID- 12777105 TI - Renormalization and destruction of 1/gamma2 tori in the standard nontwist map. AB - Extending the work of del-Castillo-Negrete, Greene, and Morrison [Physica D 91, 1 (1996); 100, 311 (1997)] on the standard nontwist map, the breakup of an invariant torus with winding number equal to the inverse golden mean squared is studied. Improved numerical techniques provide the greater accuracy that is needed for this case. The new results are interpreted within the renormalization group framework by constructing a renormalization operator on the space of commuting map pairs, and by studying the fixed points of the so constructed operator. PMID- 12777106 TI - Intermittency of energy in rapid granular shear flows. AB - Hard-disk simulations are used for two-dimensional rapid granular shear flows of circular disks between two rotating cylinders. The intermittency effects associated with the rate of the energy dissipation of collisions are studied. The statistics of intermittent signals of energy dissipation reveals that a power law governs the dynamics of rapid shear granular flows. A dynamical system approach based on the Gledzer-Ohkitani-Yamada shell model of turbulence is employed to reproduce signals for energy dissipation that are statistically consistent with those from simulations. The results suggest that rapid granular flows can be analyzed by appropriate turbulent models. PMID- 12777107 TI - Estimation of entropies and dimensions by nonlinear symbolic time series analysis. AB - Symbolic nonlinear time series analysis methods have the potential for analyzing nonlinear data efficiently with low sensitivity to noise. In symbolic nonlinear time series analysis a time series for a fixed delay is partitioned into a small number (called the alphabet size) of cells labeled by symbols, creating a symbolic time series. Symbolic methods involve computing the statistics of words made from the symbolic time series. Specifically, the Shannon entropy of the distribution of possible words for a range of word lengths is computed. The rate of increase of the entropy with word length is the metric (Kolmogorov-Sinai) entropy. Methods of computing the metric entropy for flows as well as for maps are shown. A method of computing the information dimension appropriate to symbolic analysis is proposed. In terms of this formulation, the information dimension is determined by the scaling of entropy as alphabet size is modestly increased, using the information obtained from large word length. We discuss the role of sampling time and the issue of using these methods when there may be no generating partition. PMID- 12777108 TI - Low frequency oscillations in semi-insulating GaAs: a nonlinear analysis. AB - We have observed low frequency current oscillations in a semi-insulating GaAs sample grown by low temperature molecular beam epitaxy. For this, an experimental setup proper to measure high impedance samples with small external noise was developed. Spontaneous oscillations in the current were observed for some bias conditions. Although measurements were carried out from room temperature down to liquid helium, the dynamical analysis was carried out around 200 K where the signal to noise ratio was fairly favorable. To increase the data quality we have also used a noise reduction algorithm suitably developed for nonlinear systems. We observed attractors having low embedding dimension, limit cycle bifurcations, and chaotic behavior characteristic of nonlinear dynamical processes in route to chaos. Attractor reconstruction, Poincare sections, Lyapunov exponents, and correlation dimension were also analyzed. PMID- 12777109 TI - Interpretation of heart rate variability via detrended fluctuation analysis and alphabeta filter. AB - Detrended fluctuation analysis (DFA), suitable for the analysis of nonstationary time series, has confirmed the existence of persistent long-range correlations in healthy heart rate variability data. In this paper, we present the incorporation of the alphabeta filter to DFA to determine patterns in the power-law behavior that can be found in these correlations. Well-known simulated scenarios and real data involving normal and pathological circumstances were used to evaluate this process. The results presented here suggest the existence of evolving patterns, not always following a uniform power-law behavior, that cannot be described by scaling exponents estimated using a linear procedure over two predefined ranges. Instead, the power law is observed to have a continuous variation with segment length. We also show that the study of these patterns, avoiding initial assumptions about the nature of the data, may confer advantages to DFA by revealing more clearly abnormal physiological conditions detected in congestive heart failure patients related to the existence of dominant characteristic scales. PMID- 12777110 TI - The method of variation of constants and multiple time scales in orbital mechanics. AB - The method of variation of constants is an important tool used to solve systems of ordinary differential equations, and was invented by Euler and Lagrange to solve a problem in orbital mechanics. This methodology assumes that certain "constants" associated with a homogeneous problem will vary in time in response to an external force. It also introduces one or more constraint equations. We show that these constraints can be generalized in analogy to gauge theories in physics, and that different constraints can offer conceptual advances and methodological benefits to the solution of the underlying problem. Examples are given from linear ordinary differential equation theory and from orbital mechanics. However, a slow driving force in the presence of multiple time scales contained in the underlying (homogeneous) problem nevertheless requires special care, and this has strong implications to the analytic and numerical solutions of problems ranging from celestial mechanics to molecular dynamics. (c) 2003 American Institute of Physics. PMID- 12777111 TI - Control of degenerate Hopf bifurcations in three-dimensional maps. AB - A feedback control method is proposed to create a degenerate Hopf bifurcation in three-dimensional maps at a desired parameter point. The particularity of this bifurcation is that the system admits a stable fixed point inside a stable Hopf circle, between which an unstable Hopf circle resides. The interest of this solution structure is that the asymptotic behavior of the system can be switched between stationary and quasi-periodic motions by only tuning the initial state conditions. A set of critical and stability conditions for the degenerate Hopf bifurcation are discussed. The washout-filter-based controller with a polynomial control law is utilized. The control gains are derived from the theory of Chenciner's degenerate Hopf bifurcation with the aid of the center manifold reduction and the normal form evolution. PMID- 12777112 TI - Complete synchronizability of chaotic systems: a geometric approach. AB - Synchronizability of chaotic systems is studied in this contribution. Geometrical tools are used to understand the properties of vector fields in affine systems. The discussion is focused on synchronizability of chaotic systems with equal order. The analysis is based on the synchronous behavior of all states of the master/slave system (complete synchronization). We state sufficient and necessary conditions for complete synchronizability which are based on controllability and observability of nonlinear affine systems. In this sense, the synchronizability is studied for complete synchronization via state feedback control. PMID- 12777113 TI - Chaotic mixing in a torus map. AB - The advection and diffusion of a passive scalar is investigated for a map of the 2-torus. The map is chaotic, and the limit of almost-uniform stretching is considered. This allows an analytic understanding of the transition from a phase of constant scalar variance (for short times) to exponential decay (for long times). This transition is embodied in a short superexponential phase of decay. The asymptotic state in the exponential phase is an eigenfunction of the advection-diffusion operator, in which most of the scalar variance is concentrated at small scales, even though a large-scale mode sets the decay rate. The duration of the superexponential phase is proportional to the logarithm of the exponential decay rate; if the decay is slow enough then there is no superexponential phase at all. PMID- 12777114 TI - A secure communication scheme based on the phase synchronization of chaotic systems. AB - Phase synchronization of chaotic systems with both weak and strong couplings has recently been investigated extensively. Similar to complete synchronization, this type of synchronization can also be applied in secure communications. We develop a digital secure communication scheme that utilizes the instantaneous phase as the signal transmitted from the drive to the response subsystems. Simulation results show that the scheme is difficult to be broken by some traditional attacks. Moreover, it operates with a weak positive conditional Lyapunov exponent in the response subsystem. PMID- 12777116 TI - Space-time complexity in Hamiltonian dynamics. AB - New notions of the complexity function C(epsilon;t,s) and entropy function S(epsilon;t,s) are introduced to describe systems with nonzero or zero Lyapunov exponents or systems that exhibit strong intermittent behavior with "flights," trappings, weak mixing, etc. The important part of the new notions is the first appearance of epsilon-separation of initially close trajectories. The complexity function is similar to the propagator p(t(0),x(0);t,x) with a replacement of x by the natural lengths s of trajectories, and its introduction does not assume of the space-time independence in the process of evolution of the system. A special stress is done on the choice of variables and the replacement t-->eta=ln t, s- >xi=ln s makes it possible to consider time-algebraic and space-algebraic complexity and some mixed cases. It is shown that for typical cases the entropy function S(epsilon;xi,eta) possesses invariants (alpha,beta) that describe the fractal dimensions of the space-time structures of trajectories. The invariants (alpha,beta) can be linked to the transport properties of the system, from one side, and to the Riemann invariants for simple waves, from the other side. This analog provides a new meaning for the transport exponent mu that can be considered as the speed of a Riemann wave in the log-phase space of the log-space time variables. Some other applications of new notions are considered and numerical examples are presented. PMID- 12777117 TI - The spatial logistic map as a simple prototype for spatiotemporal chaos. AB - A spatial extension of the logistic map-termed spatial logistic map-is found to display the same basic universality classes as the commonly studied diffusively coupled logistic lattice despite being vastly simpler. By analyzing the escape rates and the Lyapunov spectra it is shown that the main attractors of the spatial logistic map are stable and hence that it is a good candidate for serving as a prototype for the class of coupled map lattices which it is a part of. The spatial logistic map is then employed to provide an analytical derivation for the recently discovered linear scaling of the wavelength under increasing coupling ranges. PMID- 12777115 TI - Increased phase synchronization of spontaneous calcium oscillations in epileptic human versus normal rat astrocyte cultures. AB - Stochastic synchronization analysis is applied to intracellular calcium oscillations in astrocyte cultures prepared from epileptic human temporal lobe. The same methods are applied to astrocyte cultures prepared from normal rat hippocampus. Our results indicate that phase-repulsive coupling in epileptic human astrocyte cultures is stronger, leading to an increased synchronization in epileptic human compared to normal rat astrocyte cultures. PMID- 12777118 TI - Breaking of space-time symmetries in modulated traveling waves. AB - In this paper, we investigate the structure of traveling waves whose amplitude is spatially and/or temporally modulated. In particular, in the case of short wavelength modulations, we completely describe the breaking of symmetry induced by such modulations and their dynamical consequences. PMID- 12777119 TI - Nonlinear oscillations in marine hydroids. AB - Irregular oscillations in a colony of marine hydroids Podocoryne carnea were investigated. Quantitative characteristics were obtained as a result of long term (10-12 h) monitoring of oscillations at arbitrary sites. The sliding window spectra as well as the pulse-to-pulse dynamics argue the transient chaotic behavior of hydroid colony. The significant change of amplitudes and frequencies in intact colony oscillations after feeding and long sustained oscillations of stolons separated from colony suggest that the irregular activity could be determined by the network of pacemakers residing in stolon wall cells. These are influenced mechanically by the amount of digesting food and/or by chemical action of nutrients inside the stolon lumen. The possible correlation of these oscillations which can evoke Ca(2+) waves in stolon wall cells is discussed. PMID- 12777120 TI - Numerical analysis for a discontinuous rotation of the torus. AB - In this paper, we study a class of piecewise rotations on the square. While few theoretical results are known about them, we numerically compute box-counting dimensions, correlation dimensions and complexity of the symbolic language produced by the system. Our results seem to confirm a conjecture that the fractal dimension of the exceptional set is two, as well as indicate that the dynamics on it is not ergodic. We also explore a relationship between the piecewise rotations and discretized rotations on lattices Z(2n). PMID- 12777121 TI - Bifurcation in kinetic equation for interacting Fermi systems. AB - The recently derived nonlocal quantum kinetic equation for dense interacting Fermi systems combines time derivatives with finite time stepping known from the logistic mapping. This continuous delay differential equation is a consequence of the microscopic delay time representing the dynamics of the deterministic chaotic system. The responsible delay time is explicitly calculated and discussed for short-range correlations. As a novel feature oscillations in the time evolution of the distribution function itself appear and bifurcations up to chaotic behavior occur. The temperature and density conditions are presented where such oscillations and bifurcations arise indicating an onset of phase transition. PMID- 12777122 TI - Food chain chaos due to transcritical point. AB - Chaotic dynamics of a classical prey-predator-superpredator ecological model are considered. Although much is known about the behavior of the model numerically, very few results have been proven analytically. A new analytical result is obtained. It is demonstrated that there exists a subset on which a singular Poincare map generated by the model is conjugate to the shift map on two symbols. The existence of such a Poincare map is due to two conditions: the assumption that each species has its own time scale ranging from fast for the prey to slow for the superpredator, and the existence of transcritical points, leading to the classical mathematical phenomenon of Pontryagin's delay of loss of stability. This chaos generating mechanism is new, neither suspected in abstract form nor recognized in numerical experiments in the literature. PMID- 12777123 TI - Introduction: Nonlinear localized modes. PMID- 12777124 TI - Localization in physical systems described by discrete nonlinear Schrodinger-type equations. AB - Following a short introduction on localized modes in a model system, namely the discrete nonlinear Schrodinger equation, we present explicit results pertaining to three different physical systems described by similar equations. The applications range from the Raman scattering spectra of a complex electronic material through intrinsic localized vibrational modes, to the manifestation of an abrupt and irreversible delocalizing transition of Bose-Einstein condensates trapped in two-dimensional optical lattices, and to the instabilities of localized modes in coupled arrays of optical waveguides. PMID- 12777125 TI - Wave scattering by discrete breathers. AB - We present a theoretical study of linear wave scattering in one-dimensional nonlinear lattices by intrinsic spatially localized dynamic excitations or discrete breathers. These states appear in various nonlinear systems and present a time-periodic localized scattering potential for plane waves. We consider the case of elastic one-channel scattering, when the frequencies of incoming and transmitted waves coincide, but the breather provides with additional spatially localized ac channels whose presence may lead to various interference patterns. The dependence of the transmission coefficient on the wave number q and the breather frequency Omega(b) is studied for different types of breathers: acoustic and optical breathers, and rotobreathers. We identify several typical scattering setups where the internal time dependence of the breather is of crucial importance for the observed transmission properties. PMID- 12777126 TI - Dissipative discrete breathers: periodic, quasiperiodic, chaotic, and mobile. AB - The properties of discrete breathers in dissipative one-dimensional lattices of nonlinear oscillators subject to periodic driving forces are reviewed. We focus on oscillobreathers in the Frenkel-Kontorova chain and rotobreathers in a ladder of Josephson junctions. Both types of exponentially localized solutions are easily obtained numerically using adiabatic continuation from the anticontinuous limit. Linear stability (Floquet) analysis allows the characterization of different types of bifurcations experienced by periodic discrete breathers. Some of these bifurcations produce nonperiodic localized solutions, namely, quasiperiodic and chaotic discrete breathers, which are generally impossible as exact solutions in Hamiltonian systems. Within a certain range of parameters, propagating breathers occur as attractors of the dissipative dynamics. General features of these excitations are discussed and the Peierls-Nabarro barrier is addressed. Numerical scattering experiments with mobile breathers reveal the existence of two-breather bound states and allow a first glimpse at the intricate phenomenology of these special multibreather configurations. PMID- 12777128 TI - Cooling nonlinear lattices toward energy localization. AB - We describe the energy relaxation process produced by surface damping on lattices of classical anharmonic oscillators. Spontaneous emergence of localized vibrations dramatically slows down dissipation and gives rise to quasistationary states where energy is trapped in the form of a gas of weakly interacting discrete breathers. In one dimension, strong enough on-site coupling may yield stretched-exponential relaxation which is reminiscent of glassy dynamics. We illustrate the mechanism generating localized structures and discuss the crucial role of the boundary conditions. For two-dimensional lattices, the existence of a gap in the breather spectrum causes the localization process to become activated. A statistical analysis of the resulting quasistationary state through the distribution of breathers' energies yield information on their effective interactions. PMID- 12777127 TI - One-dimensional "turbulence" in a discrete lattice. AB - We study a one-dimensional discrete analog of the von Karman flow, widely investigated in turbulence. A lattice of anharmonic oscillators is excited by both ends in order to create a large scale structure in a highly nonlinear medium, in the presence of a dissipative term proportional to the second order finite difference of the velocities, similar to the viscous term in a fluid. In a first part, the energy density is investigated in real and Fourier space in order to characterize the behavior of the system on a local scale. At low amplitude of excitation the large scale structure persists in the system but all modes are however excited and exchange energy, leading to a power law spectrum for the energy density, which is remarkably stable against changes in the model parameters, amplitude of excitation, or damping. In the spirit of shell models, this regime can be described in terms of interacting scales. At higher amplitude of excitation, the large scale structure is destroyed and the dynamics of the system can be viewed as resulting from the creation, interaction, and decay of localized excitations, the discrete breathers, the one-dimensional equivalents of vortices in a fluid. The spectrum of the energy density is well described by the spectrum of the breathers, and shows an exponential decay with the wave vector. Due to this exponential behavior, the spectrum is dominated by the most intense breathers. In this regime, the probability distribution of the increments of velocity between neighboring points is remarkably similar to the experimental results of turbulence and can be described by distributions deduced from nonextensive thermodynamics as in fluids. In a second part the power dissipated in the whole lattice is studied to characterize the global behavior of the system. Its probability distribution function shows non-Gaussian fluctuations similar to the one exhibited recently in a large class of "inertial systems," i.e., systems that cannot be divided into mesoscopic regions which are independent. The properties of the nonlinear excitations of the lattice provide a partial understanding of this behavior. PMID- 12777129 TI - Breathers and thermal relaxation in Fermi-Pasta-Ulam arrays. AB - Breather stability and longevity in thermally relaxing nonlinear arrays depend sensitively on their interactions with other excitations. We review numerical results for the relaxation of breathers in Fermi-Pasta-Ulam arrays, with a specific focus on the different relaxation channels and their dependence on the interparticle interactions, dimensionality, initial condition, and system parameters. PMID- 12777130 TI - If "discrete breathers" is the answer, what is the question? AB - Intense work on discrete breathers or intrinsic localized modes in recent years has revealed a wealth of new properties of classical energy localization. Relaxation and mobility in particular may be two of the critical links with biomolecular processes. We review some of the basic discrete breather properties that we think are pertinent to biomolecules and make conjectures as to their possible biological utility. PMID- 12777131 TI - Selected topics in lattice dynamics: a critical review (including breathers). AB - We present a review of lattice dynamics to provide the underpinnings for the study of nonlinear localized modes, the so-called breathers. After a historical survey we address the following topics: harmonic theories, anharmonic perturbation theory, self-consistent theories, classical simulation techniques, path-integral theories, realistic crystal potentials, and intrinsic localized modes. We discuss both static and dynamic properties of crystals, e.g., neutron and x-ray scattering. We do not consider transport properties. Throughout, our emphasis is on discussing the major advances in the field and citing the appropriate references. Our aim is to achieve clarity and simplicity for readers who wish to move on to the study of breathers. We have made a special effort to set up the language and notation that is generally accepted in the field. In order to acquaint the reader with the techniques used in lattice dynamics we have analyzed a number of key problems in detail including a comparison with the available experimental data. PMID- 12777132 TI - Discrete quantum breathers: what do we know about them? AB - The knowledge about discrete quantum breathers, accumulated during the last two decades, is reviewed. "Prehistory" of the problem is described and some important properties differentiating localized and extended vibrational modes are outlined. The state of art of our understanding of the principal features of the quantum discrete breathers is presented. PMID- 12777133 TI - Discrete breathers in classical ferromagnetic lattices with easy-plane anisotropy. AB - Discrete breathers (nonlinear localized modes) have been shown to exist in various nonlinear Hamiltonian lattice systems. This paper is devoted to the investigation of a classical d-dimensional ferromagnetic lattice with easy plane anisotropy. Its dynamics is described via the Heisenberg model. Discrete breathers exist in such a model and represent excitations with locally tilted magnetization. They possess energy thresholds and have no analogs in the continuum limit. We are going to review the previous results on such solutions and also to report new results. Among the new results we show the existence of a big variety of these breather solutions, depending on the respective orientation of the tilted spins. Floquet stability analysis has been used to classify the stable solutions depending on their spatial structure, their frequency, and other system parameters, such as exchange interaction and local (single-ion) anisotropy. PMID- 12777134 TI - Collision properties of quasi-one-dimensional spin wave solitons and two dimensional spin wave bullets. AB - Collision properties of quasi-one-dimensional spin wave envelope solitons propagating in narrow ferrite film waveguides and of two-dimensional self-focused spin wave packets (spin wave bullets) propagating in wide ferrite film samples are studied both experimentally and numerically. The experiments, performed by means of a space- and time-resolved Brillouin light scattering technique, have shown that quasi-one-dimensional spin wave envelope solitons retain their shapes after collisions, while two-dimensional spin wave bullets are destroyed in collisions. The experiments have also shown that the introduction of a fixed phase shift between the colliding envelope solitons leads to a qualitative change in their interaction at the collision point. Numerical modeling of head-on collisions of nonlinear spin wave packets based on two different approaches provides a good qualitative description of the observed collision phenomena. PMID- 12777135 TI - Study of intrinsic localized vibrational modes in micromechanical oscillator arrays. AB - Intrinsic localized modes (ILMs) have been observed in micromechanical cantilever arrays, and their creation, locking, interaction, and relaxation dynamics in the presence of a driver have been studied. The micromechanical array is fabricated in a 300 nm thick silicon-nitride film on a silicon substrate, and consists of up to 248 cantilevers of two alternating lengths. To observe the ILMs in this experimental system a line-shaped laser beam is focused on the 1D cantilever array, and the reflected beam is captured with a fast charge coupled device camera. The array is driven near its highest frequency mode with a piezoelectric transducer. Numerical simulations of the nonlinear Klein-Gordon lattice have been carried out to assist with the detailed interpretation of the experimental results. These include pinning and locking of the ILMs when the driver is on, collisions between ILMs, low frequency excitation modes of the locked ILMs and their relaxation behavior after the driver is turned off. PMID- 12777136 TI - Imaging of discrete breathers. AB - In this paper I review our experiments on visualization of discrete breathers (intrinsic localized modes) in nonlinear lattices made of Josephson junctions. Properties of Josephson junctions and arrays made of such junctions are discussed in the Introduction. The visualization technique based on low temperature laser scanning microscopy (LSM) is described in detail. Images of discrete breathers in Josephson junction arrays of various geometries are presented. Possible further experiments that can be done using LSM technique are envisioned. PMID- 12777137 TI - Resonant breather states in Josephson coupled systems. AB - A review of diverse resonant effects appearing in weakly dissipative Josephson coupled systems in the presence of inhomogeneous dynamic localized state (discrete breather) is given. As particular examples I discuss the resonant interaction of breather states with linear electromagnetic excitations (EEs) in dc driven Josephson junction ladders and a single plaquette containing three Josephson junctions. Such resonant interaction manifests itself by resonant steps and various sharp switchings (voltage jumps) in the current-voltage characteristics. Moreover, the resonant interaction leads to an increase of breather dynamical complexity, e.g., enlargement of the breather core, low symmetry or quasiperiodic breather states. I show that the application of an external magnetic field allows to tune the resonant interaction, and correspondingly to increase (or decrease) the height of the resonant steps, to change the stability of the breather states. PMID- 12777138 TI - Discrete breathers in Josephson arrays. AB - Since the original proposal of 1996 by Floria et al. [Europhys. Lett. 36, 539 (1996)] of intrinsic localization in Josephson ladders, many efforts have been devoted to the theoretical, numerical, and experimental study of such dynamical states in Josephson arrays. Such efforts have already produced around 20 papers on the subject. In this article we will try to review the basic aspects of the physics of discrete breathers in Josephson arrays. PMID- 12777139 TI - Discrete solitons in inhomogeneous waveguide arrays. AB - The existence and dynamical properties of discrete solitons in inhomogeneous waveguide arrays with a Kerr nonlinearity are studied in two different configurations. First we investigate the effect of a longitudinal periodic modulation of the coupling strength on the dynamics of discrete solitons. It is shown that resonances of internal modes of the soliton with the longitudinal structure may lead to soliton oscillations and decay. Second we study the existence and stability of discrete solitons in arrays exhibiting a linear variation of the waveguide effective index in the transverse direction. We find that resonant coupling between conventional discrete solitons and linear Wannier Stark states leads to the formation of so-called hybrid discrete solitons. PMID- 12777140 TI - Intrinsic localized modes in photonic crystal circuits. AB - A discussion of intrinsic localized modes (ILMs) in photonic crystals is given. Emphasis is placed on ILMs in photonic crystal waveguides formed from Kerr nonlinear materials and photonic crystal circuits formed by joining nonlinear photonic crystal waveguides into networks. Work on stationary and propagating ILMs in nonlinear waveguides and waveguide junctions is reviewed. In addition, some discussion is given on the optical bistability of the barrier and junction geometries and on recent results on two-dimensional ILMs in Kerr nonlinear photonic crystals. PMID- 12777141 TI - Discrete nonlinear dynamics of weakly coupled Bose-Einstein condensates. AB - The dynamics of a Bose-Einstein condensate trapped in a periodic potential is governed by a discrete nonlinear equation. The interplay/competition between discreteness (introduced by the lattice) and nonlinearity (due to the interatomic interaction) manifests itself on nontrivial dynamical regimes which disappear in the continuum (translationally invariant) limit, and have been recently observed experimentally. We review some recent efforts on this highly interdisciplinary field, with the goal of stimulating interexchanges among the communities of condensed matter, quantum optics, and nonlinear physics. PMID- 12777142 TI - Spatial resonator solitons. AB - Spatial solitons can exist in various kinds of nonlinear optical resonators with and without amplification. In the past years different types of these localized structures such as vortices, bright, dark solitons, and phase solitons have been experimentally shown to exist. Many links appear to exist to fields different from optics, such as fluids, phase transitions, or particle physics. These spatial resonator solitons are bistable and due to their mobility suggest schemes of information processing not possible with the fixed bistable elements forming the basic ingredient of traditional electronic processing. The recent demonstration of existence and manipulation of spatial solitons in semiconductor microresonators represents a step in the direction of such optical parallel processing applications. We review some proof of principle soliton experiments on slow systems, and describe in more detail the experiments on semiconductor resonator solitons which are aimed at applications. PMID- 12777143 TI - Damping of persistent oscillations of quadratic optical solitons. AB - We investigate the dynamics of optical soliton formation in media with quadratic nonlinearity under conditions of long-living oscillations produced by the soliton's internal modes. We compare the predictions of the second-order perturbation approach, combining it with the energy conservation law, with the direct numerical simulations using the transparent boundary conditions. We demonstrate that these two approaches correlate well and describe the nonlinear radiation damping of the internal modes. PMID- 12777144 TI - TG-51: experience from 150 institutions, common errors, and helpful hints. AB - The Radiological Physics Center (RPC) is a resource to the medical physics community for assistance regarding dosimetry procedures. Since the publication of the AAPM TG-51 calibration protocol, the RPC has responded to numerous phone calls raising questions and describing areas in the protocol where physicists have had problems. At the beginning of the year 2000, the RPC requested that institutions participating in national clinical trials provide the change in measured beam output resulting from the conversion from the TG-21 protocol to TG 51. So far, the RPC has received the requested data from approximately 150 of the approximately 1300 institutions in the RPC program. The RPC also undertook a comparison of TG-21 and TG-51 and determined the expected change in beam calibration for ion chambers in common use, and for the range of photon and electron beam energies used clinically. Analysis of these data revealed two significant outcomes: (i) a large number (approximately 1/2) of the reported calibration changes for photon and electron beams were outside the RPC's expected values, and (ii) the discrepancies in the reported versus the expected dose changes were as large as 8%. Numerous factors were determined to have contributed to these deviations. The most significant factors involved the use of plane parallel chambers, the mixing of phantom materials and chambers between the two protocols, and the inconsistent use of depth-dose factors for transfer of dose from the measurement depth to the depth of dose maximum. In response to these observations, the RPC has identified a number of circumstances in which physicists might have difficulty with the protocol, including concerns related to electron calibration at low energies (R50<2 cm), and the use of a cylindrical chamber at 6 MeV electrons. In addition, helpful quantitative hints are presented, including the effect of the prescribed lead filter for photon energy measurements, the impact of shifting the chamber depth for photon depth-dose measurements, and the impact of updated stopping-power data used in TG-51 versus that used in TG-21, particularly for electron calibrations. PMID- 12777145 TI - Computed tomography localization of radiation treatment delivery versus conventional localization with bony landmarks. AB - A computed tomography (CT) scanner was installed in the linear accelerator room (Primatom) at Morristown. Since June 2000, we have been providing prostate, lung, and liver cancer patients with fusion of CT and linac radiation treatment. This paper describes our registration methods between planning and treatment CT images, and compares treatment localization by CT versus conventional localization by bony landmarks such as portal imaging. For image registration, we printed out beforehand the beam's eye view of the treatment fields. Prostate tumor volume from each Primatom CT slice was mapped on the printouts, and the necessary isocenter shift relative to the skin marks was deduced. No port film was necessary for our Primatom patients. For ten patients we generated digitally reconstructed radiographs (DRRs) with bone contrast from the CT scans, and deduced the required shift as the difference between the DRRs of the Primatom CT versus the planning CT. This represented the best observable shift should portal imaging be employed. Shift from bony landmark significantly correlated with the Primatom CT shift. Positioning adjustment based on bony anatomy was generally in the same direction as the CT shift for individual patient, but frequently did not go far enough. Our study confirmed that prostate organ motion relative to the bones has an average length of 4.7 mm (with standard deviation of 2.7 mm), and indicated the superiority of CT versus conventional bony structure (such as portal imaging) localization. PMID- 12777146 TI - Behavior of the Siemens Virtual Wedge following an interruption to beam delivery. AB - Investigations were made into the beam profile shape and dose delivered by the Siemens Virtual Wedge trade mark under standard operational conditions compared with those following delivery interruption on two Siemens Primus linear accelerators (Type 7445 and 8067) running different versions of control software (7.2 and 7.0, respectively). The shape of the Virtual Wedge trade mark profiles was found to be unaffected by beam delivery interruption. An increase in the dose delivered to the central axis was found when delivery was interrupted and subsequently resumed using information recorded in a recall data file on one of the accelerators. This dose increase was attributed to a difference in delivered monitor units recorded in the recall data file compared to those displayed on the linear accelerator control console. PMID- 12777148 TI - Characterization of an in vivo diode dosimetry system for clinical use. AB - An in vivo dosimetry system that uses p-type semiconductor diodes with buildup caps was characterized for clinical use on accelerators ranging in energy from 4 to 18 MV. The dose per pulse dependence was investigated. This was done by altering the source-surface distance, field size, and wedge for photons. The off axis correction and effect of changing repetition rate were also investigated. A model was developed to fit the measured two-dimensional diode correction factors. PMID- 12777147 TI - Differences in electron beam dosimetry using two commercial ionization chambers and the TG-21 protocol: another reason to switch to TG-51. AB - Two of the most popular dosimetry systems used for calibration of megavoltage photon and electron beams in radiation therapy are (i) cylindrical Farmer-type chambers in liquid water and (ii) Holt Memorial parallel-plate chambers in clear polystyrene. Since implementation of the AAPM TG-21 calibration protocol, the Radiological Physics Center (which uses the Farmer in-water system) has compared machine calibrations on two occasions with those of Memorial Sloan-Kettering Cancer Center (which uses the Holt in-polystyrene system). Two years post publication of the TG-51 protocol, 70% of the clinics monitored by the RPC still use TG-21. Seventeen photon beams from cobalt-60 to 18 MV and 31 electron beams from 6 to 20 MeV were compared using the TG-21 protocol. These data represent the most comprehensive comparison of the two most popular systems in use. Based on the average percent difference, the two systems yielded the same absorbed dose to water at the reference point in phantom to within 1.5% for both modalities. No energy dependence was evident in the results; however, a systematic average percent difference between photons and electrons was seen, with the Farmer in water system consistently predicting a dose 1.3% lower for electrons than the Holt in-polystyrene system. For photons both systems predicted the same dose to within 0.3% on average. When a physicist converts from TG-21 to TG-51, these data may be of assistance in explaining unexpected changes in output that are different from previously published values. Implementation of the TG-51 protocol should eliminate any of the observed differences in electron beam dosimetry between the two dosimetry systems because the Holt system cannot be used with TG 51. PMID- 12777149 TI - Radiation safety issues with high activities of liquid I-125: techniques and experience. AB - The handling of a liquid radioactive source is a procedure that is uncommon for the average clinical medical physicist. A newly approved treatment device utilizes high activities of liquid I-125 solution as the source of radiation. The radiation safety issues and our experience utilizing high activity liquid I-125 sources are presented. To date we have treated 22 patients with infused activities ranging up to 26.8 GBq (724 mCi). The careful manipulation of such solutions is important to maintain a safe environment for the patients and the involved medical staff. PMID- 12777150 TI - A fast, independent dose check of HDR plans. AB - High dose rate (HDR) brachytherapy often involves optimization routines to calculate the dwell times and positions of a radioactive source along specified applicator paths. These routines optimize the dwells in such a way as to deliver the prescribed dose at one or more points while satisfying various constraints. The importance of independently verifying the doses calculated by the optimization software prior to treatment delivery has been recognized in various works, and is a requirement of various regulatory agencies. Most previous methods are specific to particular treatment configurations, or require a full replanning of the case. In this work we describe an in-house software which provides an independent verification of dose calculations in less than 3 min, which adds negligible additional waiting time for the patient, regardless of the number of applicators, paths of the applicators, or complexity of the dwell times and positions. In order to verify errors which may occur between the planning and delivery stages, the verification code directly uses the treatment file used to control the HDR afterloader to compute the dose. Since this file references the source positions in the frame of reference of the catheters, an algorithm is described to convert these positions to Cartesian coordinates. We validate the code for various arbitrary cases ranging from a single catheter to complex multicatheter plans, and show results for various clinical plans. The maximum discrepancy observed for these clinical plans is 2%. PMID- 12777151 TI - Stereotactic radiotherapy in the treatment of ocular melanoma: a noninvasive eye fixation aid and tracking system. AB - Ocular melanoma is frequently treated using brachytherapy implants (such as 125I and 60Co plaques or 184Ta wire), surgery, or external beam radiotherapy using small 60Co beams, high energy x-rays, or proton therapy. The last technique, though very expensive, provides improved dose distributions and dose localizations in the treatment of tumours adjacent to critical normal tissues. The technique of fractionated stereotactic radiotherapy is now being used at an increasingly large number of centers in the treatment of lesions in the brain, and the head and neck. This article describes the successful extension of the stereotactic technique to the treatment of ocular melanoma: an eye fixation aid is attached to a noninvasive, relocatable Gill-Thomas-Cosman head frame together with a simple eye-movement tracking system. PMID- 12777152 TI - Neutron and capture gamma along the mazes of linear accelerator vaults. AB - Neutron and photon dose equivalents at various points along the mazes of the vaults for two 15 MV linear accelerators were measured. The measurements were made with the machines set at various gantry angles with and without the scattering phantom, and with the collimators set at the maximum and the minimum field sizes. Neutron dose equivalent measurements were made for five other accelerator vaults. Empirical equations were used to fit the dose data at points along the center of the maze, at 1 m above floor level, with the primary radiation beam pointing downward. It is reported here that both the capture gamma and the neutron dose attenuations along the maze are in agreement with the literature. The neutron dose is dependent on the square root of the ratio of the cross-sectional areas of the inner maze entrance and the maze. The tenth value distance (T(N)) is proportional to the square root of the cross-sectional area of the maze. PMID- 12777153 TI - Predictability of electron cone ratios with respect to linac make and model. AB - In the past, the Radiological Physics Center (RPC) has developed standard sets of photon depth-dose and wedge-factor data, specific to the make, model, and wedge design of the linear accelerator (linac). In this paper, the RPC extends the same concept to electron-cone ratios. Since 1987, the RPC has measured and documented cone-ratio (CR) values during on-site dosimetry review visits to institutions participating in National Cancer Institute cooperative clinical trials. Data have been collected for approximately 500 electron beams from a wide spectrum of linac models. The analysis presented in this paper indicates that CR values are predictable to 2% to 3% (two standard deviations) for a given make and model of linac with a few exceptions. The analysis also revealed some other interesting systematics. For some models, such as the Varian Clinac 2500 and the Elekta/Philips SL18, SL20, and SL25, CR values were nearly identical for cone sizes 15 cm x 15 cm (or 14 cm x 14 cm) and 20 cm x 20 cm across the range of available energies. Certain models of the same make of linac, such as the Mevatron MD, KD, and 6700 series models or the Clinac 2100 and 2300 models, exhibited indistinguishable CRs. Irrespective of linac model, two consistent general trends were observed: namely, an increase in CR value with incident beam energy for cone sizes smaller than 10 cm x 10 cm and a decrease with energy for cone sizes larger than 10 cm x 10 cm. These data are valuable to the RPC as a quality assurance remote-monitoring tool to identify potential dosimetry errors. The physics community will also find the data useful in several ways: as a redundant check for clinical values in use, to validate the values measured during commissioning of new machines or to ensure consistency of values measured during annual quality assurance procedures. PMID- 12777154 TI - Reimbursement versus effort in medical physics practice in radiation oncology. AB - The changes in health care reimbursement have the potential to affect the availability of quality medical physicist service in patient care. A survey was conducted by the AAPM Professional Council and the ACMP to collect cost information for special medical physics consultation, CPT4-77370 and continuing medical physics, CPT4-77336. The data collected from the survey was compared to current reimbursement schemes for a number of special procedures. Under varying reimbursement schemes, the costs of the medical physics services provided cannot be recaptured by the institution. It remains important for medical physicists to assess our utilization of resources and allocation to each of the services we provide and to understand the implications of policy changes at the federal and local levels. PMID- 12777156 TI - "Tag--we're it". PMID- 12777157 TI - Robert C. Atkins, MD, 1930-2003. In memoriam. PMID- 12777158 TI - Toxic metals and antioxidants: Part II. The role of antioxidants in arsenic and cadmium toxicity. AB - Exposure to toxic metals has become an increasingly recognized source of illness worldwide. Both cadmium and arsenic are ubiquitous in the environment, and exposure through food and water as well as occupational sources can contribute to a well-defined spectrum of disease. The symptom picture of arsenic toxicity is characterized by dermal lesions, anemia, and an increased risk for cardiovascular disease, diabetes, and liver damage. Cadmium has a significant effect on renal function, and as a result alters bone metabolism, leading to osteoporosis and osteomalacia. Cadmium-induced genotoxicity also increases risk for several cancers. The mechanisms of arsenic- and cadmium-induced damage include the production of free radicals that alter mitochondrial activity and genetic information. The metabolism and excretion of these heavy metals depend on the presence of antioxidants and thiols that aid arsenic methylation and both arsenic and cadmium metallothionein-binding. S-adenosylmethionine, lipoic acid, glutathione, selenium, zinc, N-acetylcysteine (NAC), methionine, cysteine, alpha tocopherol, and ascorbic acid have specific roles in the mitigation of heavy metal toxicity. Several antioxidants including NAC, zinc, methionine, and cysteine, when used in conjunction with standard chelating agents, can improve the mobilization and excretion of arsenic and cadmium. PMID- 12777159 TI - Giardiasis: pathophysiology and management. AB - Giardia, a common human parasite, can cause significant morbidity; however, natural medicine has great potential to influence the course of Giardia infection. The most beneficial way to treat giardiasis naturally may be through a combination approach, utilizing both nutritional interventions and phytotherapeutic agents. Nutritional intervention aims to reduce the acute symptoms of Giardia and help clear the infection. This can best be achieved by consuming a whole-food based, high-fiber, diet that is low in fat, lactose, and refined sugars. Additionally, ingestion of probiotics and wheat germ assists in parasite clearance. Numerous medicinal herbs show promise in the treatment of giardiasis. Berberine-containing herbs, garlic, and the Ayurvedic formulation Pippali rasayana currently have the most clinical evidence supporting their use. Blending the nutritional interventions and phytotherapeutic agents outlined in this article can minimize Giardia symptomatology and aid clearance of the parasite, without significant ill effects. As such, this therapeutic strategy should be considered the first-line approach. Antibiotic use may best be reserved for cases that fail to respond to initial treatment with natural measures. PMID- 12777160 TI - Probiotics in health maintenance and disease prevention. AB - Probiotic microflora display numerous health benefits beyond providing basic nutritional value. They cooperatively maintain a delicate balance between the gastrointestinal tract and immune system. When this balance is disrupted, disease and inflammation result. Inflammation and over stimulation of the immune system by pathogenic bacteria are competitively inhibited by mucosal adherence of normal beneficial microflora. A healthy gastrointestinal tract with adequate mucus production and appropriate bacterial colonization prevents the overgrowth of pathogenic bacteria, modulates disease processes, and prevents widespread inflammatory disorders. The understanding of the function of probiotics in the maintenance of health and their importance in preventing disease serves to enhance the overall health of patients. With increasing understanding that beneficial microbes are required for health maintenance and disease prevention, probiotics may be commonly used as a therapeutic tool by health care practitioners in the not-too-distant future. This review presents a review of probiotics in health maintenance and disease prevention. PMID- 12777161 TI - Cervical dysplasia: early intervention. AB - Cervical cancer is the second-most common cancer in young women and is one of the most common causes of cancer deaths among women, particularly in minorities and in impoverished countries. Cervical dysplasia, a premalignant lesion that can progress to cervical cancer, is caused primarily by a sexually transmitted infection with an oncogenic strain of the human papillomavirus (HPV). Not all women with the virus develop cervical dysplasia or cervical cancer. It has been postulated there are multiple host factors that contribute to progression of disease. Many of these factors, such as nutrient deficiencies, can be reversed, which will result in regression of dysplastic lesions. Studies have shown dietary intervention and nutrient supplementation to be effective in preventing cervical cancer. Additionally, local escharotic treatment combined with systemic treatment shows significant potential in reducing dysplasia. Recent advances in vaccination technology demonstrate the effectiveness of an HPV vaccine. The vaccine, however, may have many social and cost-prohibiting limitations, as well as health side effects. PMID- 12777162 TI - Evaluation of the effects of Neptune Krill Oil on the management of premenstrual syndrome and dysmenorrhea. AB - PRIMARY OBJECTIVE: To evaluate the effectiveness of Neptune Krill Oil (NKO) for the management of premenstrual syndrome and dysmenorrhea. SECONDARY OBJECTIVE: To compare the effectiveness of NKO for the management of premenstrual syndrome and dysmenorrhea with that of omega-3 fish oil. METHODS/DESIGN: Double-blind, randomized clinical trial. SETTING: Outpatient clinic. PARTICIPANTS: Seventy patients of reproductive age diagnosed with premenstrual syndrome according to the Diagnostic and Statistical Manual of Mental Disorders, Third Edition, Revised (DSM-III-R). INTERVENTIONS: Treatment period of three months with either NKO or omega-3 fish oil. OUTCOME MEASURES: Self-Assessment Questionnaire based on the American College of Obstetricians and Gynecologists (ACOG) diagnostic criteria for premenstrual syndrome and dysmenorrhea and number of analgesics used for dysmenorrhea. RESULTS: In 70 patients with complete data, a statistically significant improvement was demonstrated among baseline, interim, and final evaluations in the self assessment questionnaire (P < 0.001) within the NKO group as well as between-group comparison to fish oil, after three cycles or 45 and 90 days of treatment. Data analysis showed a significant reduction of the number of analgesics used for dysmenorrhea within the NKO group (comparing baseline vs. 45- vs. 90-day visit). The between-groups analysis illustrated that women taking NKO consumed significantly fewer analgesics during the 10-day treatment period than women receiving omega-3 fish oil (P < 0.03). CONCLUSION: Neptune Krill Oil can significantly reduce dysmenorrhea and the emotional symptoms of premenstrual syndrome and is shown to be significantly more effective for the complete management of premenstrual symptoms compared to omega-3 fish oil. PMID- 12777163 TI - Pellagra may be a rare secondary complication of anorexia nervosa: a systematic review of the literature. AB - Pellagra is a nutritional wasting disease attributable to a combined deficiency of tryptophan and niacin (nicotinic acid). It is characterized clinically by four classic symptoms often referred to as the four Ds: diarrhea, dermatitis, dementia, and death. Prior to the development of these symptoms, other nonspecific symptoms insidiously manifest and mostly affect the dermatological, neuropsychiatric, and gastrointestinal systems. A review of the literature reveals several case reports describing pellagra in patients with anorexia nervosa. The most common features of pellagra in patients with anorexia nervosa are cutaneous manifestations such as erythema on sun-exposed areas, glossitis, and stomatitis. Health care providers might consider a trial of 150-500 mg niacin if anorexic patients exhibit these cutaneous findings. Pellagra can be diagnosed if cutaneous symptoms resolve within 24-48 hours after oral niacin administration. To further corroborate a diagnosis of pellagra in anorexic patients, specific 24-hour urine tests for niacin metabolites and 5-hydroxy indole-acetic acid could be run prior to treatment with niacin being instituted. Other factors, such as mycotoxins, excessive dietary leucine intake (although not in anorexia), estrogens and progestogens, carcinoid syndrome, and various medications, might also lead to the development of pellagra. Although pellagra appears to be a rare, yet possible secondary complication of anorexia nervosa, it should be considered in the work-up of patients who exhibit cutaneous manifestations subsequent to sunlight exposure. PMID- 12777164 TI - Cimicifuga racemosa. Monograph. PMID- 12777165 TI - Humulus lupus. Monograph. PMID- 12777166 TI - Betaine. Monograph. PMID- 12777167 TI - X-ray structure of a putative reaction intermediate of 5-aminolaevulinic acid dehydratase. AB - The X-ray structure of yeast 5-aminolaevulinic acid dehydratase, in which the catalytic site of the enzyme is complexed with a putative cyclic intermediate composed of both substrate moieties, has been solved at 0.16 nm (1.6 A) resolution. The cyclic intermediate is bound covalently to Lys(263) with the amino group of the aminomethyl side chain ligated to the active-site zinc ion in a position normally occupied by a catalytic hydroxide ion. The cyclic intermediate is catalytically competent, as shown by its turnover in the presence of added substrate to form porphobilinogen. The findings, combined with those of previous studies, are consistent with a catalytic mechanism in which the C-C bond linking both substrates in the intermediate is formed before the C-N bond. PMID- 12777178 TI - The expression of cyclin-dependent kinase inhibitors p15, p16, p21, and p27 during ovarian follicle growth initiation in the mouse. AB - BACKGROUND: Cyclins regulate the cell cycle in association with cyclin dependent kinases (CDKs). CDKs are under inhibitory control of cyclin dependent kinase inhibitors (CDKIs). METHOD: In this study we tested the expression of CDKIs p15, p16, p21 and p27 by immunohistochemistry to determine the role of CDKIs in the initiation of primordial follicle growth. Ovaries were collected from 60-day-old cycling B6D2F1/J mice (n = 16). RESULTS: Expression of p15, p16, p21 and p27 did not vary in granulosa and theca cells by the follicle stage. However, p16 staining was stronger (++) in the oocytes of all primordial, and 57.4 +/- 3.1% of primary follicles compared to the remaining primary and more advanced follicles (+). Interestingly, primary follicles with weaker (+) oocyte staining for p16 had significantly larger mean follicle diameter compared to the primary and primordial follicles with stronger (++) oocyte staining (55.6 +/- 2.1 vs. 32.0 +/ 1.0 and 26.5 +/- 0.7 microm, respectively, p < 0.0001). This difference in follicle diameter was mainly due to a larger mean oocyte diameter (primary follicles, stronger vs. weaker, 19.6 +/- 0.6 vs. 31.5 +/- 1.4 microm, p < 0.0001). Oocytes of atretic follicles showed stronger staining with all four CDKIs. CONCLUSIONS: These preliminary findings suggest that the initiation of oocyte growth, which seems to lead follicle growth, is associated with diminished p16 expression in the mouse ovary. Further studies are needed to investigate the factors that regulate the expression of p16 in the oocyte, which might also govern the initiation of primordial follicle growth. PMID- 12777179 TI - Effects of SERM (selective estrogen receptor modulator) treatment on growth and proliferation in the rat uterus. AB - BACKGROUND: Selective estrogen receptor modulators (SERMs) have been developed in order to create means to control estrogenic effects on different tissues. A major drawback in treatment of estrogen receptor (ER) positive breast cancer with the antagonist tamoxifen (TAM) is its agonistic effect in the endometrium. Raloxifene (RAL) is the next generation of SERMs where the agonistic effect on the endometrium has been reduced. METHODS: The aim of the present study was to determine the effect of SERM treatment on the uterus, as assessed by proliferation markers and several factors involved in uterine growth. Ovariectomized (ovx) rats were treated with estradiol (E2), tamoxifen (TAM), RAL, ICI182780 (ICI) or vehicle (OVX-controls). We studied the effects on mRNA levels of the growth hormone (GH) receptor, insulin-like growth factor-I (IGF-I), ERalpha and ERbeta. In addition, by immunohistochemistry the proliferation markers PCNA and Ki-67, as well as ERalpha and ERbeta, were detected. RESULTS: The uterine weight of the rats treated with E2 or TAM was increased as compared to OVX-controls. The uterine GH-receptor mRNA level was highest in the E2 treated animals. In ICI treated rats no GH-receptor mRNA could be detected. The IGF-I mRNA level increased 16-fold in uteri of the TAM treated group and 9-fold in the E2 treated rats as compared to OVX-controls. The ERalpha mRNA level was increased in the E2 treated rats, while the ERbeta mRNA level was increased after TAM treatment. The proliferation, as assessed by PCNA, was lowest in ICI treated animals. CONCLUSIONS: The uterine wet weight, the LE height and the GH-receptor mRNA levels showed similar patterns, indicating that GH is involved in the regulation of uterine weight. Tamoxifen, which has been related to increased incidence of endometrial carcinoma in women, dramatically increased IGF-I mRNA levels in rat uterus. Since proliferation was not higher in TAM and E2 treated rats than in OVX controls, this assay of simple, early proliferation does not give the full explanation of why TAM should enhance the risk of developing endometrial cancer. PMID- 12777180 TI - Extension of adrenocortical carcinoma into the right atrium--echocardiographic diagnosis: a case report. AB - BACKGROUND: Adrenocortical carcinoma is a rare, highly malignant tumor. Cardiac involvement of the tumor is very rare. Echocardiography facilitates the evaluation of the cardiac involvement of the tumor. CASE PRESENTATION: We describe a patient with an adrenal tumor. Transthoracic echo showed its extension into the right atrium. Accordingly, a combined abdominal and cardiac operation was performed, monitored by transesophageal echocardiography. CONCLUSION: This case highlights the importance of echocardiography in revealing the cardiac involvement by this tumor and in planning the operative procedure. PMID- 12777181 TI - Acute health effects after accidental exposure to styrene from drinking water in Spain. AB - OBJECTIVES: We studied subjective health symptoms in a population accidentally exposed to high styrene concentrations in drinking tap water. The contamination occurred during the reparation of a water tank. METHODS: Residents of 27 apartments in two buildings using the contaminated water were contacted. A questionnaire on subjective symptoms was administered to 84 out of 93 persons living in the apartments at the time of the accident. Styrene concentration was measured in samples of water collected two days after the accident. The means of exposure associated with appearance of symptoms were examined through case control analyses. RESULTS: Styrene in water reached concentrations up to 900 microg/L. Symptoms were reported by 46 persons (attack rate 55 %). The most frequent symptoms were irritation of the throat (26%), nose (19%), eyes (18%) and the skin (14%). General gastrointestinal symptoms were observed with 11% reporting abdominal pain and 7% diarrhea. The factors most strongly associated with symptoms were drinking tap water (OR = 7.8, 95% CI 1.3-48), exposure to vapors from the basement (OR = 10.4, 2.3-47) and eating foods prepared with tap water (OR = 8.6, 1.9-40). All residents in the ground floor reported symptoms. CONCLUSIONS: This accidental contamination led to very high styrene concentrations in water and was related to a high prevalence of subjective symptoms of the eyes, respiratory tract and skin. Similar exposures have been described in workers but not in subjects exposed at their residence. Various gastrointestinal symptoms were also observed in this population probably due to a local irritative effect. PMID- 12777182 TI - Hip disability and osteoarthritis outcome score (HOOS)--validity and responsiveness in total hip replacement. AB - BACKGROUND: The aim of the study was to evaluate if physical functions usually associated with a younger population were of importance for an older population, and to construct an outcome measure for hip osteoarthritis with improved responsiveness compared to the Western Ontario McMaster osteoarthritis score (WOMAC LK 3.0). METHODS: A 40 item questionnaire (hip disability and osteoarthritis outcome score, HOOS) was constructed to assess patient-relevant outcomes in five separate subscales (pain, symptoms, activity of daily living, sport and recreation function and hip related quality of life). The HOOS contains all WOMAC LK 3.0 questions in unchanged form. The HOOS was distributed to 90 patients with primary hip osteoarthritis (mean age 71.5, range 49-85, 41 females) assigned for total hip replacement for osteoarthritis preoperatively and at six months follow-up. RESULTS: The HOOS met set criteria of validity and responsiveness. It was more responsive than WOMAC regarding the subscales pain (SRM 2.11 vs. 1.83) and other symptoms (SRM 1.83 vs. 1.28). The responsiveness (SRM) for the two added subscales sport and recreation and quality of life were 1.29 and 1.65, respectively. Patients 66 years of age (range 67-85) (Pain SRM 2.60 vs. 1.97, other symptoms SRM 3.0 vs. 1.60, activity of daily living SRM 2.51 vs. 1.52, sport and recreation function SRM 1.53 vs. 1.21 and hip related quality of life SRM 1.95 vs. 1.57). CONCLUSION: The HOOS 2.0 appears to be useful for the evaluation of patient-relevant outcome after THR and is more responsive than the WOMAC LK 3.0. The added subscales sport and recreation function and hip related quality of life were highly responsive for this group of patients, with the responsiveness being highest for those younger than 66. PMID- 12777183 TI - Awareness of vaccination status and its predictors among working people in Switzerland. AB - BACKGROUND: Adult vaccination status may be difficult to obtain, often requiring providers to rely on individual patient recall. To determine vaccination status awareness and the sociodemographic predictors of awareness for tetanus, hepatitis A and B, tick born encephalitis (TBE) and influenza vaccination. METHODS: Multivariate analyses were used to evaluate a questionnaire survey of 10,321 employees (4070 women and 6251 men aged 15-72 years) of two companies in Switzerland. RESULTS: Among 10,321 respondents, 75.5% reported knowing their tetanus vaccination status, 64.1% hepatitis A, 61.1% hepatitis B, 64.3% TBE and 71.9% influenza. Between 1 in 4 and 1 in 3 employees were not aware of their vaccination status. Differences in awareness for the five vaccinations considered correlated with gender and language. These differences persisted in multivariate analyses. CONCLUSION: Women employees, German-speaking employees and employees who paid more attention to their diet were more often aware of their vaccination status. A more reliable and readily accessible data source for vaccination status is needed in order to capitalize on opportunities to update vaccinations among Swiss employees. PMID- 12777186 TI - The role of stool DNA analysis in the early detection and screening of colorectal cancer. PMID- 12777184 TI - Interactions within the mammalian DNA methyltransferase family. AB - BACKGROUND: In mammals, epigenetic information is established and maintained via the postreplicative methylation of cytosine residues by the DNA methyltransferases Dnmt1, Dnmt3a and Dnmt3b. Dnmt1 is required for maintenance methylation whereas Dnmt3a and Dnmt3b are responsible for de novo methylation. Contrary to Dnmt3a or Dnmt3b, the isolated C-terminal region of Dnmt1 is catalytically inactive, despite the presence of the sequence motifs typical of active DNA methyltransferases. Deletion analysis has revealed that a large part of the N-terminal domain is required for enzymatic activity. RESULTS: The role played by the N-terminal domain in this regulation has been investigated using the yeast two-hybrid system. We show here the presence of an intra-molecular interaction in Dnmt1 but not in Dnmt3a or Dnmt3b. This interaction was confirmed by immunoprecipitation and was localized by deletion mapping. Furthermore, a systematic analysis of interactions among the Dnmt family members has revealed that DNMT3L interacts with the C-terminal domain of Dnmt3a and Dnmt3b. CONCLUSIONS: The lack of methylating ability of the isolated C-terminal domain of Dnmt1 could be explained in part by a physical interaction between N- and C terminal domains that apparently is required for activation of the catalytic domain. Our deletion analysis suggests that the tertiary structure of Dnmt1 is important in this process rather than a particular sequence motif. Furthermore, the interaction between DNMT3L and the C-terminal domains of Dnmt3a and Dnmt3b suggests a mechanism whereby the enzymatically inactive DNMT3L brings about the methylation of its substrate by recruiting an active methylase. PMID- 12777187 TI - 14th EORTC-NCI-AACR Symposium on Molecular Targets and Cancer Therapeutics. PMID- 12777185 TI - High accuracy genotyping directly from genomic DNA using a rolling circle amplification based assay. AB - BACKGROUND: Rolling circle amplification of ligated probes is a simple and sensitive means for genotyping directly from genomic DNA. SNPs and mutations are interrogated with open circle probes (OCP) that can be circularized by DNA ligase when the probe matches the genotype. An amplified detection signal is generated by exponential rolling circle amplification (ERCA) of the circularized probe. The low cost and scalability of ligation/ERCA genotyping makes it ideally suited for automated, high throughput methods. RESULTS: A retrospective study using human genomic DNA samples of known genotype was performed for four different clinically relevant mutations: Factor V Leiden, Factor II prothrombin, and two hemochromatosis mutations, C282Y and H63D. Greater than 99% accuracy was obtained genotyping genomic DNA samples from hundreds of different individuals. The combined process of ligation/ERCA was performed in a single tube and produced fluorescent signal directly from genomic DNA in less than an hour. In each assay, the probes for both normal and mutant alleles were combined in a single reaction. Multiple ERCA primers combined with a quenched-peptide nucleic acid (Q-PNA) fluorescent detection system greatly accellerated the appearance of signal. Probes designed with hairpin structures reduced misamplification. Genotyping accuracy was identical from either purified genomic DNA or genomic DNA generated using whole genome amplification (WGA). Fluorescent signal output was measured in real time and as an end point. CONCLUSIONS: Combining the optimal elements for ligation/ERCA genotyping has resulted in a highly accurate single tube assay for genotyping directly from genomic DNA samples. Accuracy exceeded 99 % for four probe sets targeting clinically relevant mutations. No genotypes were called incorrectly using either genomic DNA or whole genome amplified sample. PMID- 12777188 TI - Activity of the Raf kinase inhibitor BAY 43-9006 in patients with advanced solid tumors. PMID- 12777189 TI - Overview of preoperative and postoperative therapy for colorectal cancer: the European and United States perspectives. AB - Surgery is the primary modality for cure in patients with localized colorectal cancer. However, despite potential curative surgery, the risk of recurrence is high. In colon cancer, the role of adjuvant chemotherapy with 5-fluorouracil (5 FU) and leucovorin (LV) is now established in stage III disease. The benefit of adjuvant treatment in stage II disease is likely to be small, and studies performed thus far have been generally underpowered to detect what might be a clinically significant effect on survival. Whereas bolus scheduling of 5-FU and LV is favored in North America, infusion of 5-FU/LV is preferred in Europe. Indeed, infused 5-FU/LV may be a safer partner with new drugs such as oxaliplatin and irinotecan. Oral fluoropyrimidines are attractive agents that might one day replace parenteral 5-FU. In rectal cancer, postoperative combined chemoradiation was recommended as standard practice in stages II and III disease. Despite a lack of randomized data demonstrating clinical benefit, preoperative chemoradiation has been increasingly used in patients with T3 disease in North America. However, preoperative radiation therapy is more frequently used in Europe. There are discrepancies in pathologic reporting of circumferential resection margin involvement and lymph node status between the United States and Europe. Standardized reporting with improved preoperative imaging would allow patients with truly early-stage disease to undergo more conservative management and be spared the morbidity and mortality of unnecessary adjuvant or neoadjuvant treatment. PMID- 12777190 TI - Locoregional strategies for colorectal hepatic metastases. AB - Hepatic colorectal metastases present a challenging problem in patients with recurrent colorectal cancer. Twenty percent of patients with recurrence will have liver metastases as a component of their disease, and only 10% of these patients will have isolated liver metastases that are resectable. Although systemic chemotherapy alone has not proven efficacious in the treatment of liver metastases, a number of options exist in managing these lesions. For those that are resectable, surgery remains the optimal treatment, with an expected 5-year survival rate of 33%-39% based on several large series. Hepatic artery chemotherapy is another adjunct treatment in patients undergoing resection and may further improve survival. This benefit may be even more pronounced when combined with systemic chemotherapy. Newer-generation agents are likely to further improve results. More recently, new therapeutic modalities have been used to treat unresectable lesions. These include hepatic artery chemotherapy and ablative techniques such as radiofrequency ablation and cryoablation. This article will highlight the data regarding hepatic resection for colorectal metastases, describe the rationale for and efficacy of hepatic arterial chemotherapy in the postoperative adjuvant setting and in unresectable liver disease, and review the current literature describing ablative techniques in the treatment of liver metastases. PMID- 12777191 TI - Impact of locoregional approaches to liver metastases in patients with colorectal cancer. PMID- 12777192 TI - Sensitivity and specificity of a stool DNA multitarget assay panel for the detection of advanced colorectal neoplasia. AB - Colorectal cancer is the second-leading cause of cancer death. New noninvasive options for screening capable of diagnosing cancer at an early stage are needed to improve compliance and reduce mortality. This study was designed to provide an estimate of the sensitivity and specificity of a multitarget assay panel (MTAP) of stool DNA changes. Eighty patients with advanced colorectal neoplasia and 212 control subjects provided stool samples before colonoscopy. Patients with hereditary colorectal cancer syndromes were excluded. The MTAP included 21 specific mutations in the adenomatous polyposis coli (APC), p53, and K-ras genes, a microsatellite instability marker (BAT-26), and a marker of abnormal apoptosis (DNA Integrity Assay). All samples were analyzed in the clinical laboratory at EXACT Sciences. Multitarget assay panel detected 33 of 52 patients (63.5%, 95% confidence interval [CI], 49.0%-76.4%) with invasive colorectal cancer, including 26 of 36 (72.2%) with node-negative disease (American Joint Committee on Cancer [AJCC] stage I/II) and 7 of 16 (43.7%) with advanced disease (AJCC stage III/IV). Sixteen of 28 patients (57.1%; 95% CI, 37.2%-75.5%) with advanced adenomas (lesions containing high-grade dysplasia, villous adenomas, or tubular adenomas > 1 cm in size) were detected, including 6 of 7 (85.7%) with high-grade dysplasia and 10 of 21 (47.6%) with other advanced adenomas. Specificity was 96.2% (95% CI, 92.7%-98.4%) in patients with either no colorectal lesions or diminutive polyps. Multitarget assay panel has better sensitivity than that reported with use of Hemoccult(R) II in fecal occult blood testing, with similar specificity. Sensitivity appeared to be equally high for patients with node-negative and advanced disease, as well as for advanced adenomas. This study contained a disproportionately high number of distal cancers and, as such, may not be representative of results in proximal lesions. Although a prospective study in an average-risk population is needed to validate these findings, MTAP may offer an important noninvasive option for population-based screening. PMID- 12777193 TI - Unusual sites of colorectal cancer metastasis. AB - We present 2 separate cases of adenocarcinoma of the colon with metastasis to the chin and the bladder, both of which are unusual sites of colorectal cancer metastasis. Patient 1 is a 77-year-old man who was diagnosed with adenocarcinoma of the colon, American Joint Committee on Cancer (AJCC) T4 N0 M0 (stage II), and underwent a right hemicolectomy. Fourteen months later he developed a firm 2.5-cm mass involving the chin. Excisional biopsy revealed moderately differentiated adenocarcinoma, consistent with the known colon primary tumor. Patient 2 is a 75 year-old man who was diagnosed with AJCC T3 N1 M0 (stage III) adenocarcinoma of the colon and underwent sigmoid colectomy. Ten years later, he was found to have transitional cell carcinoma involving retroperitoneal nodes with no identifiable bladder or ureteral primary, for which he received chemotherapy. Eighteen months following this diagnosis, he developed hematuria and was found to have metastatic colon adenocarcinoma involving the bladder. Details of both patient cases are presented here. PMID- 12777194 TI - A trial of adjuvant therapy in colorectal cancer: the VICTOR trial. PMID- 12777195 TI - Thrombolytic Therapy in Acute Ischemic Stroke. AB - The use of intravenous thrombolytic therapy for the treatment of patients with acute ischemic stroke is now approved in the United States, Canada, Germany, and the European Union. Guidelines published in 1996 from the American Heart Association and American Academy of Neurology committees recommended intravenous administration of recombinant tissue-plasminogen activator (rt-PA) (0.9 mg/kg; maximum of 90 mg) given in a 10% bolus, followed by an infusion lasting 60 minutes, to patients within 3 hours of onset of ischemic stroke. The recommendations stipulate that a computed tomography scan before the infusion should not show major infarction, mass effect, edema, or hemorrhage. Yet, only a small fraction of eligible patients (< 5%) have received rt-PA during the 7 years since its approval in the United States. Although effective, thrombolysis carries an important risk (5% to 10%) of brain hemorrhage and edema that can prove fatal. Many physicians and medical centers are not presently equipped or willing to give thrombolytic drugs for stroke treatment. PMID- 12777196 TI - Cerebral Venous Thrombosis. AB - Because of its wide range of presentations, its highly variable mode of onset, its numerous causes, and its unpredictable outcome, cerebral venous thrombosis (CVT) remains a diagnostic and therapeutic challenge. Treatment of CVT consists primarily of symptomatic treatment of seizures and intracranial hypertension, antithrombotics, and etiologic treatment whenever possible. Heparin remains the first line of treatment for CVT; although its systematic use remains debated, recent studies have confirmed its safety even in patients with large hemorrhagic infarctions. The addition of local thrombolysis is indicated for patients with clinical worsening related to extension of the venous thrombosis, despite adequate anticoagulation and optimal symptomatic and etiologic treatment. In contrast to arterial stroke, complete recovery of prolonged or severe neurologic deficit is possible, justifying initiation of anticoagulation and eventually thrombolysis, even when the clinical situation seems desperate. New techniques using mechanical devices disrupting the clot may be used in addition to thrombolysis in rare cases. Ventricular drainage is indicated in cases of cerebellar infarction or deep venous thrombosis associated with hydrocephalus. Decompressive craniotomy may be performed acutely in patients with untractable intracranial hypertension and herniation. PMID- 12777197 TI - Hypoperfusion and Its Augmentation in Patients with Brain Ischemia. AB - Control of hypertension is a well-established goal of the primary and secondary prevention of stroke. However, management of blood pressure in the setting of acute brain ischemia is complicated by the possible effect of blood pressure changes on cerebral perfusion. In acute stroke, patients may have an ischemic penumbra of brain tissue, which has impaired perfusion but which is not irreversibly damaged. The ischemic penumbra may be salvaged with reperfusion. Lowering of blood pressure in this setting, however, would hasten the progression of the penumbra to infarction. With the exception of patients treated with thrombolytic agents, blood pressure reduction is not recommended in acute ischemic stroke for this reason. Preliminary studies suggest that there may be a role for interventions to elevate blood pressure as a treatment for acute stroke patients. Despite interest in induced hypertension as a treatment of stroke dating back to the 1950s, this practice has not achieved widespread use owing to concerns about potential adverse effects such as intracerebral hemorrhage, cerebral edema, and myocardial ischemia. It is commonly used, however, to treat patients with threatened cerebral ischemia due to vasospasm after subarachnoid hemorrhage. Until future studies clarify the effectiveness of induced hypertension in stroke treatment, maintaining adequate blood pressure and fluid volume is recommended for patients with acute ischemic stroke, particularly if the neurologic deficits are fluctuating or the patient has persistent large vessel occlusive disease. PMID- 12777198 TI - Need for Continued Use of Anticoagulants After Intracerebral Hemorrhage. AB - One of the unsolved dilemmas in cerebrovascular disease is the ideal timing to restart anticoagulant therapy (ACT) safely in patients who have suffered an intracranial hemorrhage and have an indication to continue ACT. No randomized studies with evidence-based data are available to settle this issue. Only a few reports have been published; their shortcomings include small numbers of patients evaluated retrospectively, a broad spectrum of indications for ACT, different types of hemorrhage, and variable methods of reversing and restarting ACT. Despite this variability, most reports agree that 1) ACT has to be immediately reversed to decrease the risk of hemorrhage progression; 2) a period between 1 and 2 weeks appears sufficient to allow for management and monitoring of the hemorrhage off ACT; and 3) ACT can be safely restarted after the period off of treatment. A physician confronting a patient with intracranial hemorrhage and the need for ACT faces a situation of individually focused clinical decision making. The problem rests in balancing the risks of a worsening or recurring hemorrhage on one side, and the risk of thromboembolism on the other. From available information and personal experience, an unvalidated risk stratification can be proposed to categorize patients in low- and high-risk groups for hemorrhagic complications and embolic phenomena, assigning them to 5 or 15 days off ACT, respectively. Intermediate-risk patients can have ACT restarted in 5 to 15 days from the onset of the intracranial hemorrhage. Anticoagulation should be immediately reversed. Fresh frozen plasma is the standard treatment in most institutions. Prothrombin complex concentrate is an alternative, but issues of availability make its use impractical. The use of recombinant factor VIIa is an attractive new option. PMID- 12777199 TI - Aortic Source of Brain Embolism. AB - Aortic arch atheroma has more recently been identified as an independent risk factor for ischemic stroke. Initially, this was a result of careful autopsy observations, then followed by a series of in vivo studies in which aortic arch atheroma was identified by transesophageal echocardiography. The association of aortic arch atheroma with ischemic stroke is most likely causal, given that the stroke risk increases with increasing thickness of arch atheroma. There is quite a sharp increase in stroke risk for atheroma of 4 mm or greater compared with lesser thicknesses. The clinical diagnosis is suggested when transient ischemic attack or ischemic stroke has occurred in which no obvious cardiac or arterial source of embolism is found. The presence of aortic arch atheroma is usually detected by transesophageal echocardiography and sometimes by magnetic resonance imaging or computed tomography. There is uncertainty about clinical management, particularly for secondary prevention. Options include the use of antiplatelet agents, anticoagulants, thrombolysis, or surgery. The latter two options have only been described rarely in case reports. Of the less invasive approaches, combination antiplatelet therapy with aspirin and clopidogrel is favored, or the use of warfarin. The Aortic arch Related Cerebral Hazard (ARCH) trial is being conducted to determine which of these is more effective in minimizing a composite outcome cluster of ischemic stroke, intracranial hemorrhage, myocardial infarction, peripheral embolism, or vascular death. Other more general management strategies should include reasonably aggressive risk factor control with blood pressure and lipid-lowering therapies and, if indicated, careful diabetic control. PMID- 12777201 TI - Patent Foramen Ovale and Stroke. AB - Observational data from prospective and retrospective trials indicate that a patent foramen ovale (PFO) is associated with the risk of ischemic stroke. The mechanism involved is presumed to be paradoxical embolism from a venous thrombus that travels via the PFO to the systemic circulation causing an ischemic stroke. Primary stroke prevention data for patients with a PFO are nonexistent. Given the substantial prevalence of PFO in the total population (~ 25% to 30%), a primary prevention study may not be feasible. However, whether targeted primary prevention for patients with PFOs of certain morphologic characteristics (eg, larger size, greater degree of shunt) would be possible remains undefined. Given the large number of asymptomatic subjects, no therapy is currently recommended. The best treatment modality to prevent recurrent stroke in patients with PFO has not been defined. There are four major treatment choices: surgical closure, percutaneous device closure, medical therapy with anticoagulants, and medical therapy with antiplatelet agents. Regarding medical therapy, the Patent Foramen Ovale in Cryptogenic Stroke Study has demonstrated that antiplatelet and anticoagulant therapies are of equal benefit in preventing recurrent adverse events. Although closure of the PFO, either surgical or percutaneous, may further reduce the event rates, this remains to be demonstrated because no randomized trial to date has compared PFO closure with medical therapy. PMID- 12777200 TI - Brain Embolism Monitoring with Transcranial Doppler Ultrasound. AB - Embolism is considered to be the main mechanism leading to brain infarction today; with the introduction of sophisticated neuroimaging tools, its impact is increasingly appreciated. Transcranial Doppler ultrasound allows noninvasive monitoring of in vivo embolism. Acute stroke, internal carotid artery stenosis, several cardiac conditions, internal carotid endarterectomy, and coronary artery bypass graft surgery have been extensively monitored. These investigations and other clinical and neuroimaging studies have expanded the understanding of brain embolism; they suggest it may be appropriate to think of it as a process that occurs in the context of other hemodynamic factors. Differences have been identified among several conditions regarding the temporal profile of embolism and the characteristics of embolic particles. This article presents a brief review of brain embolism monitoring with transcranial Doppler ultrasound. PMID- 12777202 TI - Cerebrovascular Disease in Pregnancy. AB - Cerebrovascular disease in pregnancy poses challenges to clinicians because of the difficulty in diagnosing the underlying etiology as well as the potential fetal toxicity of diagnostic testing and treatment. The underlying etiology for stroke in the pregnant patient must be aggressively sought and is critical to appropriate therapy: 1) If the arterial ischemic event in a pregnant patient remains cryptogenic, then either low-dose aspirin or another antithrombotic agent should be used. 2) Unfractionated heparin, or preferably, low molecular weight heparin, should be used to treat patients with a clearly identified etiology for which there is a specific clinical indication with at risk or symptomatic doses. Warfarin may be used in occasional cases when heparin is contraindicated (heparin induced thrombocytopenia) and should be limited to the period after organogenesis in the second and third trimesters. 3) Using a heparin-warfarin-heparin alternating schedule to offset adverse events is impractical, because with each change in medications sustained anticoagulation cannot be easily obtained. PMID- 12777203 TI - Atherosclerotic Vertebral Artery Disease in the Neck. AB - Atherosclerotic lesions are very common at the origin and first few centimeters of the vertebral artery in the neck. These lesions are often missed when using noninvasive diagnostic strategies. These lesions cause transient hypoperfusion and transient ischemic attacks, characterized mostly by vestibulocerebellar symptoms. Strokes are caused by embolism from these lesions. Patients with nonstenosing vertebral artery plaques should be treated with statins and antiplatelet agents. Symptomatic patients who have had embolism from an occluded vertebral artery should be treated with heparin followed by warfarin for a period of 4 to 12 weeks. Antiplatelet agents can then be given. The optimal treatment of asymptomatic and symptomatic patients with severe vertebral artery stenosis is unclear. Statins and warfarin are recommended for patients with concurrent stenotic lesions of the contralateral extracranial vertebral artery, the intracranial vertebral or basilar arteries, or the anterior circulation arteries. Balloon angioplasty or stenting will likely prove superior to surgery in those patients with severe stenosis who do not respond to medical treatment. PMID- 12777204 TI - Hepatic clearance of apoptotic lymphocytes: simply removal of waste cells? AB - The in situ liver recognition of apoptotic lymphocytes was studied by using different sources of lymphocytes (i.e. human, rat and mouse) and animal models (i.e. rat and mouse). Lymphocytes were induced to apoptosis using 10(-2)M cycloheximide for up to 24 hours; three types of apoptosing lymphocytes, corresponding to different stages in the apoptotic process, were described: type 1 or early apoptosis, type 2 or mature apoptosis and type 3 or late/necrotic apoptosis. When livers were in situ injected with apoptotic lymphocytes enriched for type 1 (early), 2 (mature) or 3 (late/necrotic) apoptosis, they recognized and internalized apoptosing cells, with an efficiency directly dependent on the stage of the apoptotic process. The highest recognition rate, which was, in all cases, mediated by galactose- and mannose-specific receptors, was obtained with homologous apoptotic cells (i.e. rat lymphocytes and rat liver). Moreover, the drastically reduced efficiency of recognition of human or mouse apoptotic lymphocytes when injected into rat liver, suggested the involvement also of species-specific antigens. PMID- 12777205 TI - 60KDa chaperonin (HSP60) is over-expressed during colorectal carcinogenesis. AB - The aim of the present study was to evaluate the expression of the heat shock protein 60 (HSP60), a mitochondrial matrix-associated protein belonging to the chaperonin family, in colorectal adenomas and cancers, comparing them to normal colonic tissues and hyperplastic polyps. We performed both immunohistochemistry and Western blot analysis for HSP60. Immunohistochemistry resulted positive in all tubular adenomas and infiltrating adenocarcinomas. By contrast, normal tissues and hyperplastic polyps were negative. Quantitative analysis showed that tubular adenomas with different levels of dysplasia did not present statistical differences concerning HSP60 positivity. In addition, carcinomas always showed the highest expression. Western blot analysis confirmed these observations. These data suggest that HSP60 over-expression is an early event in carcinogenesis. We suspect that HSP60 plays a different role in colorectal carcinogenesis with respect to that in normal cells, which foresees its possible use as diagnostic and prognostic tools. PMID- 12777206 TI - Detection of cancer clones in human gastric adenoma by increased DNA-instability and other biomarkers. AB - An immunohistochemical differential staining of cancerous cells with anti cytidine antibody after denaturation of nuclear DNA by acid hydrolysis with 2N HCl at 30 degree C for 20 min (DNA-instability test) has been used as a marker of malignancy. The test was applied to bioptic tissues of human gastric polyp assessed histopathologically as foveolar hyperplastic polyp (13 cases), mild (58 cases), moderate (86 cases), and severe (20 cases) dysplasia, and adenocarcinomas (14 cases). The serial sections of the same tissues were also subjected to immunohistochemical staining for Ki67, p53, DNA-fragmentation factor (DFF45), and basic fibroblast growth factor (bFGF). The DNA-instability test was positive in 14 (100%) adenocarcinoma cases, 20 (100%) severe dysplasia cases, 52 (60.5%) moderate dysplasia cases, and 12 (20.7%) mild dysplasia cases, indicating malignancy. All foveolar hyperplastic polyps were negative to the DNA-instability testing. Furthermore, the percentage of glands positive in the DNA-instability test steadily increased in going from mild (10%), to moderate (40%), to severe (100%) dysplasia, and adenocarcinoma (100%). All other biological markers tested in the present study showed significantly higher values in the adenoma glands, being positive to DNA-instability testing, irrespective of the dysplasia grade, as compared to those in the adenoma glands that were negative to DNA-instability testing. Furthermore, the former values were comparable to those in adenocarcinoma. These results indicate that cancer cell clones are already present at the adenoma stages showing a positive DNA-instability test, enhanced proliferative activity, p53 mutation, induction of DFF45 and bFGF. These factors allow cancer cell proliferation, producing heterogeneous subclones due to DNA instability, enhancing their survival by escaping apoptosis, and providing abundant nutrients during the early-stage progression of gastric cancer. Based on these findings, we herein propose the concept of "procancer" (as opposed to "pre cancer") as being a unique stage during the course of carcinogenesis and cancer progression. We designate the term to cancer clones at the very early stages of malignant progression that do not show distinguishable morphological atypia but do show positive DNA-instability testing and positive staining for various biomarkers such as Ki67, p53, DFF45, and bFGF. We also define the abnormal positive staining of these biomarkers, including the DNA-instability test as "functional atypia", compared to the ordinary morphological atypia. PMID- 12777207 TI - Ki-67 immunolabeling in pre-malignant lesions and carcinoma of the prostate. Histological correlation and prognostic evaluation. AB - The antigen Ki-67, which is associated with cell proliferation, has been demonstrated to be useful in predicting the development of human tumors. The objective of this study was to evaluate the prognostic utility of this biomarker in pre-malignant and malignant lesions of the prostate. A total of 162 prostate biopsies taken from patients diagnosed for benign prostatic hyperplasia (BPH, n=49), low grade prostatic intraepithelial neoplasia (LGPIN, n=53), high grade prostatic intraepithelial neoplasia (HGPIN, n=25) and carcinoma (CAR, n=35), were studied. Immunohistochemistry for Ki-67 was carried out on all the samples and the number of labeled cells was semi-quantitatively evaluated (weak, moderate or intense). In the non-invasive lesions, the presence of Ki-67-positive cells in the luminal layer of the epithelium was evaluated qualitatively as positive or negative. The correlation between the immunolabeling for Ki-67 and the histological diagnosis showed highly significant differences between BPH and CAR, LGPIN and CAR and HGPIN and CAR, with no significant differences being found among the other groups. Analysis of the immunolabeling in luminal cells of non invasive lesions showed an increase in accordance with the increase in the degree of histological lesion, the greatest percentage being obtained in the HGPIN lesions (88.0%), with significant differences among all the groups. Bearing in mind that Ki-67 is a prognostic biomarker for cell proliferation, our results demonstrating the immunolabeling of Ki-67 in the luminal compartment of non invasive lesions having the potential to evolve to malignancy, may have prognostic implications. PMID- 12777208 TI - Biological aggressiveness evaluation in prostate carcinomas:immunohistochemical analysis of PCNA and p53 in a series of Gleason 6 (3+3) adenocarcinomas. AB - We selected 63 prostate tumors with Gleason's grade 6 (3+3), commonly showing both tubular and cribrous patterns. We compared in both patterns the expression of two of the most used biologic markers: PCNA and p53, with the aim to verify the validity of the Gleason's grading system to compare the morphologic grade with biologic aggressiveness and prognostic value. We did not find any statistical difference in the protein immunopositivity, indicating that both patterns could have identical biologic behaviour; then we confirmed the validity of Gleason's system for considering both tubular and cribrous patterns as an intermediate grade of tumoral differentiation. Moreover, we found a linear relationship between the increase of PCNA and the accumulation of mutated p53; this datum could confirm the hypothesis that p53 mutation is a late event in prostate carcinogenesis. PMID- 12777209 TI - Presence of atrial natriuretic factor in normal and hyperplastic human prostate and its relationship with oxytocin localisation. AB - In this work, we showed the presence of atrial natriuretic factor (ANF) in human prostate and compared its localisation in normal and hyperplastic conditions. ANF was localised in epithelial and stromal cells, being increased in hyperplasia, mainly in the stromal component. Moreover, we compared ANF and oxytocin positivity in the same glands, focusing on the possible relationship between the paracrine effects of these two hormones. PMID- 12777210 TI - Parkia pendula lectin as histochemistry marker for meningothelial tumour. AB - Lectins have been intensively used in histochemical techniques for cell surface characterization. These proteins are involved in several biological processes and their use as histochemical markers have been evaluated since they can indicate differences in cell surfaces. Parkia pendula lectin (PpeL) was evaluated as histochemical marker for meningothelial meningioma biopsies. Tissue slices were incubated with PpeL conjugated to horseradish peroxidase (PpeL-HRP) and Concanavalin A-HRP (ConA-HPR) and the binding visualized with diaminobenzidine and hydrogen peroxide. The lectin-tissue binding was inhibited with D-glucose. PpeL showed to be a useful tool for the characterization of meningothelial tumour and clinico-pathological diagnosis. PMID- 12777211 TI - An immunohistochemical study and review of potential markers of human intestinal M cells. AB - M cells are found in intestinal follicle associated epithelium. Studies into the physiological and pathological roles of human M cells have been hampered by the lack of well-substantiated, specific markers for these cells. A critical literature review suggests the following molecules may potentially serve as such markers: CK7, FcaR (CD89), S100, CD1a, CD21, CD23, sialyl Lewis A, and cathepsin E. Normal ileum, appendix and colorectum were studied using paraffin-embedded, formalin-fixed tissue and immunohistochemistry for these 8 markers. Cathepsin E immunohistochemistry was also performed on cases of colorectal adenocarcinoma, colorectal adenoma, colorectal hyperplastic/metaplastic polyp, lymphocytic colitis, collagenous colitis, pseudomembranous colitis and active ulcerative colitis. Of the 8 markers tested, only cathepsin E appeared to be specific to follicle associated epithelium (expressed by cells with and without M cell morphology) and follicular crypt epithelium; this specificity was limited to the colorectum. Focal epithelial expression of cathepsin E was seen in adenocarcinoma, adenoma, hyperplastic/metaplastic polyp, ulcerative colitis and pseudomembranous colitis. In conclusion, cathepsin E is a specific marker of normal colorectal follicle associated epithelium and follicular crypt epithelium though is not specific to M cells within these compartments. None of the other 7 markers studied is exclusively expressed by human M cells. PMID- 12777212 TI - Association of Maedi Visna virus with Brucella ovis infection in rams. AB - Maedi Visna Virus (MVV) is the etiological agent of a systemic disease of sheep, which causes lesions in lungs, the central nervous system, joints, and mammary glands. It has been speculated that the association with Brucella ovis may lead to the venereal shedding of the virus. In this work, samples of epididymis from ten rams positive for MVV and infected experimentally with Brucella ovis, were subjected to liquid-phase PCR, immunohistochemistry (IHC) and in situ PCR tests, aimed at identifying the pathogens in a tissue context. IHC was carried out using a monoclonal antibody raised against p28 MVV protein and a polyclonal antibody to B. ovis. Liquid phase- and in situ PCR were designed to amplify a portion of MVV proviral DNA Pol sequence. In the animals showing B. ovis-related histopathological changes, IHC clearly demonstrated a positivity for B. ovis and MVV in interstitial and epithelial ductal cells. In situ PCR assessed the presence of MVV proviral DNA in macrophages and elements inside the epithelium. The unaffected and reagent control samples constantly gave negative results. Taken together, these data demonstrate that MVV may affect ovine epididymis, apparently taking advantage of the concurrent infection by B. ovis. The tropism of MVV for the epididymal epithelial cells, may be responsible for its excretion with the semen. PMID- 12777213 TI - The autonomous innervation of the buffalo (Bubalus bubalis) testis. An immunohistochemical study. AB - The innervation pattern in the buffalo testis was determined by using histochemical and immunohistochemical methods. Nerves were concentrated in the tunica albuginea and septula testis, and did not show an uniform distribution. The tunica albuginea at the lateral and medial sides and at the free border of the testis is most densely innervated than at the epididymal border. At the cranial pole thick nerve bundles were observed between albugineal vessels and muscle bundles. Rare parenchymal nerves were found in perivascular position between seminiferous tubules and their occurrence is confined to lobules at the cranial and caudal testicular poles. An intense NPY immunoreactivity occurred in nerve bundles and in solitary varicose fibres. Nerves were concentrated in the tunica albuginea at the lateral and medial side and at the free border of the testis, and in the lobules at the cranial and caudal testicular poles. Sub P immunoreactivity was occasionally detected in some thicker nerve bundles and solitary fibers, in the tunica albuginea and in the wall of blood vessels, showing a similar distribution but less intensity and density than NPY immunoreactivity. TH immunoreactivity stained nerve fibers in the buffalo testis with a distribution pattern similar to that obtained with general neuronal markers. The histochemical reaction for AchE was negative, so cholinergic fibers cannot be detected in the buffalo testis. The histochemical NADPHd reaction stained rare nitrergic nerve bundles and solitary fibers. The majority of NADPHd activity was confined to the vascular endothelium, and rarely to the interstitial Leydig cells, whereas the Sertoli and germ cells did not show any reaction. PMID- 12777214 TI - An immunohistochemical study of endocrine cells in the pancreas of the Red bellied frog (Bombina orientalis). AB - The regional distribution and frequency of pancreatic endocrine cells in the red bellied frog, Bombina orientalis, were studied by the immunohistochemical peroxidase anti-peroxidase (PAP) method using five types of specific mammalian antisera to insulin, glucagon, somatostatin, bovine pancreatic polypeptide (PP) and secretin. The frequency was calculated as the mean number of each endocrine cell type/1,000 total cells (including exocrine and endocrine cells) using an automated image analysis process. The percentage of each immunoreactive (IR) cell species to the total IR cell population was also calculated. In the pancreas of the red-bellied frog, all five endocrine cell types were demonstrated. Insulin IR cells were located in the pancreas as single cells or islet-like clusters. The latter were localized in central regions. The insulin-IR cells showed a frequency of 65.40 plus/minus 14.56/1,000 cells. Glucagon IR cells were also detected as single cells or as clusters but in the case of clusters, two distributional patterns were detected - a central core type and a marginally distributed type. They showed an abundance of 32.70 plus/minus 7.32/1,000 cells. Somatostatin-IR cells were dispersed throughout the pancreatic parenchyma as single cells, three to four cells, or clusters. The clusters were located in the marginal regions. The somatostatin-IR cell frequency was 19.40 plus/minus 6.52/1000 cells. PP-IR cells were randomly distributed throughout the pancreatic parenchyma as single cells with a frequency of 14.70 plus/minus 4.92/1,000 cells. Secretin-IR cells were demonstrated as clusters or as single cells, and as clusters they occupied the central regions. They showed a frequency of 39.60 plus/minus 10.36/1,000 cells. This is the first report of the presence of secretin-IR cells in amphibian pancreatic endocrine cells. Overall, there were 37.20 plus/minus 6.84% insulin-, 21.90 plus/minus 5.55% glucagon-, 11.60 plus/minus 4.33% somatostatin-, 8.60 plus/minus 2.72% PP- and 23.40 plus/minus 4.45% secretin-IR cells. PMID- 12777216 TI - Detection of MBL-2 gene expression in intestinal biopsies of celiac patients by in situ reverse transcription polymerase chain reaction. PMID- 12777215 TI - Demonstration of glucose-6-phosphate dehydrogenase in rat Kupffer cells by a newly-developed ultrastructural enzyme-cytochemistry. AB - Although various tissue macrophages possess high glucose-6-phosphate dehydrogenase (G6PD) activity, which is reported to be closely associated with their phagocytotic/bactericidal function, the fine subcellular localization of this enzyme in liver resident macrophages (Kupffer cells) has not been determined. We have investigated the subcellular localization of G6PD in Kupffer cells in rat liver, using a newly developed enzyme-cytochemical (copper ferrocyanide) method. Electron-dense precipitates indicating G6PD activity were clearly visible in the cytoplasm and on the cytosolic side of the endoplasmic reticulum of Kupffer cells. Cytochemical controls ensured specific detection of the enzymatic activity. Rat Kupffer cells abundantly possessed enzyme cytochemically detectable G6PD activity. Kupffer cell G6PD may play a role in liver defense by delivering NADPH to NADPH-dependent enzymes. G6PD enzyme cytochemistry may be a useful tool for the study of Kupffer cell functions. PMID- 12777217 TI - Card tricks. Getting a prescription drug discount can save you money, but you may have to earn it. Finding the right card involves shopping around and reading the fine print. PMID- 12777218 TI - A cheaper way to control high blood pressure. PMID- 12777219 TI - Fish vs. fish oil capsules. Can you skip the fish and get the same benefits from taking fish oil capsules? PMID- 12777220 TI - Hard facts about stones. Kidney stones can be painful, but most pass. PMID- 12777221 TI - The smallpox vaccine: frequently asked questions. PMID- 12777222 TI - Retirement benefits. PMID- 12777223 TI - Toil in the soil. PMID- 12777224 TI - Leave the driving to them. PMID- 12777225 TI - By the way, doctor. I'm 5-foot-8 and 170 pounds, so my BMI is 26. I exercise a lot, and no one thinks I'm overweight. But the BMI charts say I am. What do you think? PMID- 12777226 TI - By the way, doctor. Is gamma-tocopherol a better form of vitamin E than alpha tocopherol? PMID- 12777227 TI - By the way, doctor. I've had prostate cancer and am taking selenium in hopes of keeping it under control. Will it affect the absorption of vitamins? PMID- 12777229 TI - Regular alcohol consumption is good for the heart. PMID- 12777228 TI - What you need to know about calcium. Calcium is billed as the bone-building nutrient. But some experts argue that we should pay more attention to exercise and vitamin D. PMID- 12777230 TI - Older women don't get the heart medications they need. PMID- 12777231 TI - Bell's palsy. Early treatment with antiviral medications and corticosteroids may improve the chances for full recovery from this condition that suddenly paralyzes one side of the face. PMID- 12777232 TI - Marathons: blood, sweat, and cheers. Marathon running may be extreme, but there are some lessons about exercise in it for all of us. PMID- 12777233 TI - A side effect of COX-2 inhibitors. PMID- 12777234 TI - Grape juice vs. red wine. PMID- 12777235 TI - Leave no instrument (or sponge) behind. PMID- 12777236 TI - By the way, doctor. I'm 84, 5-foot-9, and weigh 160. I do an hour's exercise every day. I take a diuretic, potassium, and calcium-channel blocker. Should I also take Lipitor to avoid a heart attack? PMID- 12777237 TI - By the way, doctor. I'm 77 and have an enlarged prostate. I take Proscar once a day and saw palmetto three times a day. Yet I still have to get up two to seven times a night to urinate. What can I do to reduce the frequency of urination? PMID- 12777238 TI - Branch and foliage morphological plasticity in old-growth Thuja plicata. AB - At the Wind River Canopy Crane Facility in southeastern Washington State, USA, we examined phenotypic variation between upper- and lower-canopy branches of old growth Thuja plicata J. Donn ex D. Don (western red cedar). Lower-canopy branches were longer, sprouted fewer daughter branches per unit stem length and were more horizontal than upper-canopy branches. Thuja plicata holds its foliage in fronds, and these had less projected area per unit mass, measured by specific frond area, and less overlap, measured by silhouette to projected area ratio (SPARmax), in the lower canopy than in the upper canopy. The value of SPARmax, used as an indicator of sun and shade foliage in needle-bearing species, did not differ greatly between upper- and lower-canopy branches. We suggest that branching patterns, as well as frond structure, are important components of morphological plasticity in T. plicata. Our results imply that branches of old-growth T. plicata trees have a guerilla growth pattern, responding to changes in solar irradiance in a localized manner. PMID- 12777239 TI - Development and recovery from winter embolism in silver birch: seasonal patterns and relationships with the phenological cycle in oceanic Scotland. AB - Silver birch (Betula pendula Roth) is increasingly used in the United Kingdom for reforestation. However, recent evidence indicates that, under some circumstances, planted birch can suffer serious and repeated mortality of the apical leaders and branches, with consequent loss of apical dominance and the formation of a contorted stem. Plants from 37 seed sources of silver birch from Scotland and northern England planted at two sites were compared for several characteristics related to hydraulic architecture, vulnerability to freeze-thaw cycle induced embolism and spring recovery from winter embolism during the period 2000-2002. Phenological rhythms were also monitored in late winter-early spring to document relationships between phenology and water relations parameters. Significant differences were found across seed sources in stage of bud flushing for four dates in spring. Early flushing seed sources differed by about 1 to 2 weeks from late-flushing seed sources across the two sites. Wintertime xylem embolism in stems reached a peak of about 50 to 70% loss of xylem hydraulic conductivity, depending on the size and position of the sample shoots in the canopy. Small apical shoots were significantly more embolized than large basal shoots. Development of winter embolism was coupled to the occurrence of frost events. As percent loss of hydraulic conductivity increased during the winter, wood relative water content declined. Embolism reversal occurred rapidly in spring at the time of development of positive root pressure. No significant differences in the degree of winter embolism in 2001 were found among the three seed sources examined. The investigation was expanded in the winter-spring of 2002 to include 10 seed sources across both sites. Significant differences were found in degree of winter embolism across sites, dates and seed sources. For each date, there was a significant relationship between flushing scores and wood relative water contents across the two sites and all seed sources, suggesting that differences in time of flushing across sites and seed sources were likely caused by differences in the time of occurrence of root pressure, a necessary precondition to flushing. PMID- 12777240 TI - Branchlet nutrient concentration in hoop pine (Araucaria cunninghamii) relative to family, stable carbon and oxygen isotope ratios and growth rate in contrasting environments. AB - Genetic variation in branchlet nutrient (N, P, K, Na, Ca, Mg, Mn and Fe) concentrations and mineral concentration (sum of branchlet P, K, Na, Ca, Mg, Mn and Fe concentrations) of 8-9-year-old hoop pine (Araucaria cunninghamii Ait. ex D. Don) half-sib families was assessed for four canopy positions at a wet site (23 families) and two canopy positions at an N- and water-limiting dry site (22 families) in relation to tree growth and associated branchlet carbon (delta13C) and oxygen (delta18O) isotope composition in southeast Queensland, Australia. Branchlet nutrient and mineral concentrations varied significantly among families and with canopy position and site. Depending on the canopy position sampled, the hoop pine family effect accounted for 0 to 13.8% of the total variation in branchlet N concentration, and for 0 to 30.3% of the total variation in branchlet mineral concentration at the wet site. The corresponding values for the family effect at the dry site were 0-13.3% for branchlet N concentration and 0-25.7% for branchlet mineral concentration. There were significant variations in branchlet P, K, Ca and Mg concentrations at both sites, and these variations differed with canopy position. Relationships between family means of branchlet N concentration and tree growth or delta13C or delta18O varied with canopy position at both sites. At the wet site, there were significant positive correlations between branchlet mineral concentration in the upper-outer or upper-inner canopy and tree height (r = 0.26 and 0.37, P < 0.01) and between branchlet mineral concentration and delta13C (r = 0.24, P < 0.01) in the upper-inner canopy, and a significant negative correlation between branchlet mineral concentration and delta13C (r = 0.21, P < 0.05) in the upper-outer canopy. At the dry site, branchlet mineral concentrations in the upper-inner and upper-outer canopy were significantly correlated with branchlet delta13C (r = -0.28 and -0.51, P < 0.01), and branchlet N concentration in the upper-inner canopy was significantly correlated with tree growth (r = 0.29, P < 0.01). A significant correlation between branchlet delta18O (an index of stomatal conductance) and branchlet mineral concentration at the dry site (r = 0.39, P = 0.020) indicated that stomatal conductance might be a factor regulating the variation in branchlet mineral concentration of the hoop pine families. Both branchlet N concentration and mineral concentration at particular canopy positions assist in selecting hoop pine families with improved tree growth and N- and water-use efficiency in environments where both N deficiency and a limited water supply are major factors affecting plantation productivity. PMID- 12777242 TI - Daily shoot extension growth of peach trees growing on rootstocks that reduce scion growth is related to daily dynamics of stem water potential. AB - We studied relationships between diurnal patterns of stem water potential (PsiSTEM) and stem extension growth of the same scion cultivar growing on three rootstocks with differing size-controlling potentials. The peach trees (Prunus persica (L.) Batsch) used in this field experiment consisted of an early-maturing freestone cultivar, 'Flavorcrest,' grafted onto three different rootstocks: Nemaguard (a vigorous seed-propagated control, P. persica x P. davidiana hybrid), Hiawatha (an intermediate vigor rootstock, derived from an open pollinated seedling of a P. besseyi x P. salicina hybrid) and K-146-43 (a semi-dwarfing rootstock, P. salicina x P. persica hybrid). Diurnal patterns of PsiSTEM and stem extension growth were measured on six dates (March 29, April 12, April 26, May 10, May 24 and June 18) during the primary period of peach shoot extension growth. Rootstocks clearly affected diurnal patterns of PsiSTEM and stem extension growth. Trees on K-146-43 had the lowest midday PsiSTEM and stem extension growth. Differences among rootstocks in the amount of diurnal oscillation in PsiSTEM explained stem extension rate differences induced by the three rootstocks. The sensitivity of shoot extension growth to tree water relations tended to decrease as the season progressed and was not apparent by mid June. The results of the study indicate that water relations may play an important role in the dwarfing mechanism induced by size-controlling peach rootstocks. PMID- 12777241 TI - Light-energy processing and freezing-tolerance traits in red spruce and black spruce: species and seed-source variation. AB - Red spruce (Picea rubens Sarg.) and black spruce (Picea mariana (Mill.) B.S.P.) are genetically and morphologically similar but ecologically distinct species. We determined intraspecific seed-source and interspecific variation of red spruce and black spruce, from across the near-northern margins of their ranges, for several light-energy processing and freezing-tolerance adaptive traits. Before exposure to low temperature, red spruce had variable fluorescence (Fv) similar to black spruce, but higher photochemical efficiency (Fv/Fm), lower quantum yield, lower chlorophyll fluorescence (%), and higher thermal dissipation efficiency (qN), although the seed-source effect and the seed-source x species interaction were significant only for Fv/Fm. After low-temperature exposure (-40 degrees C), red spruce had significantly lower Fv/Fm, quantum yield and qN than black spruce, but higher chlorophyll fluorescence and relative fluorescence. Species, seed source effect, and seed-source x species interaction were consistent with predictions based on genetic (e.g., geographic) origins. Multi-temperature exposures (5, -20 and -40 degrees C) often produced significant species and temperature effects, and species x temperature interactions as a result of species-specific responses to temperature exposures. The inherent physiological species-specific adaptations of red spruce and black spruce were largely consistent with a shade-tolerant, late-successional species and an early successional species, respectively. Species differences in physiological adaptations conform to a biological trade-off, probably as a result of natural selection pressure in response to light availability and prevailing temperature gradients. PMID- 12777243 TI - Growth, photosynthetic performance and shade adaptation of rubber (Hevea brasiliensis) grown in natural shade. AB - We compared growth, photosynthetic performance and shade adaptation of rubber (Hevea brasiliensis Muell. Arg.) plants growing in natural shade (33, 55 and 77% reduction in incoming radiation) to control plants growing in full sunlight. Stem diameter and plant height, measured over a 15-month period, were greatest in plants grown in full sunlight, and both parameters decreased with increasing shade. At 7 and 14 months after planting (MAP), total plant dry mass was highest in control plants and lowest in plants in 77% shade. Expansion of the fourth leaf whorl, monitored at 5-6 MAP, was slowest in plants in 77% shade and fastest in unshaded plants, which had more leaves and higher leaf areas and inter-whorl shoot lengths. In response to increasing shade, specific leaf area increased, whereas leaf weight ratio and relative growth rate decreased. Chlorophyll a/b ratio decreased with increasing shade, indicating shade-induced partitioning of chlorophyll into light-harvesting complexes. Compared to the response in unshaded plants, CO2 assimilation saturated at lower photosynthetic photon flux densities in plants in 77% shade, with a lower upper-asymptote to the light response curve. Chlorophyll fluorescence revealed no evidence of sustained photoinhibitory damage in unshaded plants. Dynamic photoinhibition decreased with increasing shade, with the greatest depression in the ratio of variable to maximal fluorescence around midday. We conclude that shade adaptation and shade-induced reductions in dynamic photoinhibition account for the enhanced early growth of rubber in light shade. PMID- 12777244 TI - Root formation from transgenic calli of Ginkgo biloba. AB - The objective of this study was to produce Ginkgo biloba L. hairy roots for future investigations into the feasibility of producing terpenoids in differentiated cell cultures. Zygotic embryos of G. biloba were inoculated with the wild agropine-type Agrobacterium rhizogenes strain A4. Three months after bacterial infection, primordia-like nodular structures formed at the root wound sites and developed into tiny calli. Calli cultivated on hormone-free Lloyd and McCown (1980) solid medium were transferred onto Murashige and Skoog (1962) solid medium, and grew rapidly for the first few months. As browning appeared and growth slowed, calli were transferred onto Ball (1959) or White (1954) solid media. These calli became nodular with several nodules from which roots, displaying characteristic features of hairy roots, developed. Transgenic calli cultivated in agitated, hormone-free liquid media led to the formation of root meristems and root tips by a cyclic development process. Stable integration of the rolA, rolB and rolC genes into calli, mature roots and root meristem and root tip mixtures was confirmed by polymerase chain reaction (PCR). The absence of a 437 bp amplificate corresponding to the virD1 gene confirmed that there was no bacterial contamination of G. biloba tissues. The reverse transcription-PCR method was used to verify the expression of rolA and rolC genes in mature roots. Expression of rolA, rolB and rolC in root meristems and root tips was confirmed by 3' RACE-PCR analysis, which excluded amplification of possible rol gene transcripts produced by residual bacteria. This paper shows, for the first time, the feasibility of developing G. biloba roots from transformed calli. PMID- 12777245 TI - Oxidized low-density lipoprotein activates migration and degranulation of human granulocytes. AB - Oxidized low-density lipoprotein (oxLDL) has been reported as a major participant in the pathogenesis of atherosclerosis. We hypothesized that oxLDL can also interact with granulocytes during inflammatory airway diseases, such as asthma. To test the chemotactic effect of oxLDL, isolated human peripheral granulocytes were added to the upper chambers of Transwell filters and migration in response to oxLDL was determined. Cu+2-oxidized LDL stimulated neutrophil (23.4 +/- 3.2% for 100 microg/ml oxLDL versus 2.9 +/- 1.1% for buffer, P < 0.05) and eosinophil (19.3 +/- 3.5% versus 0.6 +/- 0.02% for buffer, P < 0.05) chemotaxis in a concentration-dependent manner. The magnitude of chemotaxis was dependent on the degree of LDL oxidation. Granulocyte transmigration across IL-1beta-activated human pulmonary microvascular endothelial cell monolayers was similarly stimulated by oxLDL. OxLDL activated significant degranulation of both neutrophils (100.9 +/- 9.8 versus 49.6 +/- 8.4 ng lactoferrin released/5 x 105 neutrophils for buffer, P < 0.05) and eosinophils (342 +/- 115.4 versus 85.8 +/- 30.4 ng eosinophil-derived neurotoxin/1 x 106 eosinophils for buffer, P < 0.05). Therefore, in vivo influx and oxidation of LDL may be an important mediator for the initiation of bronchial inflammation where granulocytes are recruited to the lung. PMID- 12777246 TI - Expression and localization of lung surfactant protein A in human tissues. AB - Lung surfactant protein A (SP-A) is a collectin produced by alveolar type II cells and Clara cells. It binds to carbohydrate structures on microorganisms, initiating effector mechanisms of innate immunity and modulating the inflammatory response in the lung. Reverse transcriptase-polymerase chain reaction was performed on a panel of RNAs from human tissues for SP-A mRNA expression. The lung was the main site of synthesis, but transcripts were readily amplified from the trachea, prostate, pancreas, and thymus. Weak expression was observed in the colon and salivary gland. SP-A sequences derived from lung and thymus mRNA revealed the presence of both SP-A1 and SP-A2, whereas only SP-A2 expression was found in the trachea and prostate. Monoclonal antibodies were raised against SP-A and characterized. One of these (HYB 238-4) reacted in Western blotting with both reduced and unreduced SP-A, with N-deglycosylated and collagenase-treated SP-A, and with both recombinant SP-A1 and SP-A2. This antibody was used to demonstrate SP-A in immunohistochemistry of human tissues. Strong SP-A immunoreactivity was seen in alveolar type-II cells, Clara cells, and on and within alveolar macrophages, but no extrapulmonary SP-A immunoreactivity was observed. In contrast to lung surfactant protein D (SP-D), which is generally expressed on mucosal surfaces, SP-A seems to be restricted to the respiratory system. PMID- 12777247 TI - Involvement of serum response factor isoforms in myofibroblast differentiation during bleomycin-induced lung injury. AB - Serum response factor (SRF) is a transcription factor essential for smooth muscle (SM) myogenesis. Its role in myofibroblast differentiation is, however, unknown. We studied the expression and the localization of SRF in bleomycin-induced pulmonary fibrosis, where myofibroblasts are abundant. We found that SRF levels were upregulated in bleomycin-exposed mouse lungs mainly due to de novo synthesis of SRFDelta5, a less myogenic SRF isoform. Before myofibroblast differentiation, SRF/SRFDelta5 was immunolocalized mostly in the cytoplasm of scattered fibroblasts at lesion sites. With the development of myofibroblasts, however, SRF/SRFDelta5 was found in myofibroblast nuclei. cDNA array analysis showed that SRFDelta5 and SRF induced expression of transforming growth factor-beta1, a critical factor in myofibroblast differentiation. This was accompanied by de novo expression of several inflammatory cell-specific mRNAs. The latter was confirmed by reverse transcriptase-polymerase chain reaction. Treatment of lung fibroblasts with tumor necrosis factor-alpha, which is produced early in the bleomycin model, induced SRFDelta5 expression and SRF/SRFDelta5 cytoplasmic accumulation, whereas addition of transforming growth factor-beta1 caused SRF/SRFDelta5 nuclear translocation followed by SM alpha-actin synthesis. Interleukin-4, another cytokine involved in myofibroblast differentiation, did not affect SRF or induce SRFDelta5 expression. Our studies therefore suggested a new mechanism whereby SRF and SRFDelta5 contribute to the emergence of myofibroblasts in lung injury and fibrosis. PMID- 12777248 TI - Antigen presentation by local macrophages promotes nonallergic airway responses in sensitized mice. AB - Local inflammatory responses involve relocating immune functions generated by previous immunization to confined parts of the body, and hence are presumed to reflect the prevailing systemic immune bias. To verify to what extent local antigen-presenting cells (APCs) may modulate immune inflammation, we analyzed the consequences of antigen presentation by macrophages on Th2-dependent airway inflammation in ovalbumin (OVA)-sensitized mice. In contrast to challenge with free OVA, which triggers airway eosinophilia and Th2 cell recruitment, intratracheal instillation of immortalized spleen macrophages (Mf4/4 cells), pulsed with OVA, promoted a nonallergic airway response featuring recruitment of interferon-gamma-producing Th1 cells. Combining OVA-Mf4/4 instillation with OVA inhalation strongly reduced airway eosinophilia. Inflammation repression persisted after secondary OVA challenge and depended on the antigen-presenting ability of the macrophages. Arguing against Th1-mediated counter-regulation, Th1/Th2 ratios remained unaltered in macrophage-treated/OVA-challenged mice. In contrast, levels of interleukin-4 and interleukin-13 mRNA in lung tissue CD4+ T cells were strongly downregulated, indicating a suppression of Th2 cell activation. These results document a role for local macrophages/APCs in controlling the nature and intensity of local immune inflammatory responses. The resulting segregation of systemic and local levels of immune reactivity may enable local inflammation tolerance; it is a nonallergic airway response despite systemic sensitization. PMID- 12777249 TI - Human ADAM33 messenger RNA expression profile and post-transcriptional regulation. AB - We examined transcript expression and post-transcriptional regulation of human ADAM33, a recently identified asthma gene. A detailed messenger RNA (mRNA) expression profile was obtained using Northern, reverse transcription polymerase chain reaction, and in situ hybridization analyses. ADAM33 mRNA was expressed significantly in smooth muscle-containing organs, minimally in immune organs and hematopoietic cells, and highly in repairing duodenal granulation tissue. Expression was seen in asthmatic subepithelial fibroblasts and smooth muscle but not in respiratory epithelium. In all tissues, transcripts of approximately 5 kb predominated over those of approximately 3.5 kb by 2- to 5-fold. The effect of the 3' untranslated region (UTR) on ADAM33 protein expression and maturation was examined. The presence of the 3'UTR in untagged full-length constructs promoted prodomain removal, detected as mature approximately 100 kD protein by ADAM33 reactive antibodies; in its absence, maturation was 2- to 3-fold less in HEK293 cells. His-tagged and untagged constructs lacking the 3'UTR demonstrated that lack of maturation was not a result of tag-mediated effects. Minimal maturation of ADAM33 occurred in primary lung and MRC5 fibroblasts following adenoviral mediated expression of ADAM33 lacking the 3'UTR. In contrast, prodomain removal was observed with plasmids and adenovirus encoding only the pro- and catalytic domains. Thus, the 3'UTR of ADAM33 and domains downstream of the catalytic domain regulate potential ADAM33 activity. Mechanisms of regulation of ADAM33, distinct from closely related ADAMs, thus include mRNA localization and processing and protein maturation. PMID- 12777250 TI - The role of the basolateral outwardly rectifying chloride channel in human airway epithelial anion secretion. AB - The purpose of this study was to characterize basolateral anion channels in Calu 3 and normal human bronchial epithelial cells, and their role in anion secretion. Patch clamp studies identified an outwardly rectifying Cl- channel (ORCC), which could be activated by the adenosine receptor agonist 5'-(N ethylcarboxamido)adenosine (NECA). Short-circuit current measurements revealed that NECA activates a basolateral, but not an apical, anion conductance sensitive to 4,4'-diisothiocyanatostilbene-2, 2'-disulfonic acid, and to 9 anthracenecarboxylic acid, but not to 4,4'-dinitrostilbene-2,2'-disulfonic acid. Apical membrane permeabilization studies confirmed the presence of basolateral anion channels, established their halide permeability sequence (Cl- >/= Br- >> I ), and demonstrated their outwardly rectifying nature. Experiments using H-89, forskolin, and Ht31 demonstrated that adenosine receptor dependent activation of basolateral ORCC was cAMP- and potentially A-kinase anchoring protein-dependent. Neither BAPTA-AM treatment nor basolateral Ca2+ removal had any effect on the activation of these channels. Anion replacement and 36Cl- flux studies show that Calu-3 cells primarily secrete HCO3- when stimulated with NECA, and that Cl- secretion can be stimulated by blocking basolateral ORCC, whereas normal human bronchial epithelial cells exclusively secrete Cl- under all conditions studied. We propose a novel model of anion secretion in which ORCC recycles Cl- across the basolateral membrane, allowing preferential HCO3- secretion. PMID- 12777251 TI - Aberrant lung structure, composition, and function in a murine model of Hermansky Pudlak syndrome. AB - Hermansky-Pudlak syndrome (HPS) is a genetically heterogeneous inherited disease causing hypopigmentation and prolonged bleeding times. An additional serious clinical problem of HPS is the development of lung pathology, which may lead to severe lung disease and premature death. No cure for the disease exists, and previously, no animal model for the HPS lung abnormalities has been reported. A mouse model of HPS, which is homozygously recessive for both the Hps1 (pale ear) and Hps2 (pearl) genes, exhibits striking abnormalities of lung type II cells. Type II cells and lamellar bodies of this mutant are greatly enlarged, and the lamellar bodies are engorged with surfactant. Mutant lungs accumulate excessive autofluorescent pigment. The air spaces of mutant lungs contain age-related elevations of inflammatory cells and foamy macrophages. In vivo measurement of lung hysteresivity demonstrated aberrant lung function in mutant mice. All these features are similar to the lung pathology described in HPS patients. Morphometry of mutant lungs indicates a significant emphysema. These mutant mice provide a model to further investigate the lung pathology and therapy of HPS. We hypothesize that abnormal type II cell lamellar body structure/function may predict future lung pathology in HPS. PMID- 12777252 TI - The role of regulated CFTR trafficking in epithelial secretion. AB - The focus of this review is the regulated trafficking of the cystic fibrosis transmembrane conductance regulator (CFTR) in distal compartments of the protein secretory pathway and the question of how changes in CFTR cellular distribution may impact on the functions of polarized epithelial cells. We summarize data concerning the cellular localization and activity of CFTR and attempt to synthesize often conflicting results from functional studies of regulated endocytosis and exocytosis in CFTR-expressing cells. In some instances, findings that are inconsistent with regulated CFTR trafficking may result from the use of overexpression systems or nonphysiological experimental conditions. Nevertheless, judging from data on other transporters, an appropriate cellular context is necessary to support regulated CFTR trafficking, even in epithelial cells. The discovery that disease mutations can influence CFTR trafficking in distal secretory and recycling compartments provides support for the concept that regulated CFTR recycling contributes to normal epithelial function, including the control of apical CFTR channel density and epithelial protein secretion. Finally, we propose molecular mechanisms for regulated CFTR endocytosis and exocytosis that are based on CFTR interactions with other proteins, particularly those whose primary function is membrane trafficking. These models provide testable hypotheses that may lead to elucidation of CFTR trafficking mechanisms and permit their experimental manipulation in polarized epithelial cells. PMID- 12777253 TI - A new chapter in cardiac PKC signaling studies: searching for isoform-specific molecular targets. Focus on: "isoenzyme-selective regulation of SERCA2 gene expression by protein kinase C in neonatal rat ventricular myocytes". PMID- 12777254 TI - Impaired osmotic sensation in mice lacking TRPV4. AB - The Ca2+-permeable cation channel TRPV4, which is part of the Trp family located in the circumventricular organs, is activated by cell swelling. To investigate the role of TRPV4 in osmotic sensation, we disrupted the TRPV4 gene in mice and examined the effect on osmotic metabolism. Disruption of the mouse TRPV4 gene did not influence either water intake behavior or serum osmolality. Short-term salt ingestion, however, seemed to impair the transient free water clearance. The level of serum arginine vasopressin (AVP) of TRPV4-/- mice was not significantly changed under normal conditions but was significantly increased under stimulated conditions. Incubation of brain slices with graded hyperosmolality suggested an exaggerated response of AVP secretion in TRPV4-/- mice. Thus TRPV4 in the brain may transmit a negative signal to AVP secretion similar to an inhibitory pass through the baroregulatory system. Thus, in the regulation of serum osmolality, TRPV4 is a swell-activated channel that appears to play a role in reversion toward hyposmolality. PMID- 12777255 TI - Differential regulation of junctional complex assembly in renal epithelial cell lines. AB - Several signaling pathways that regulate tight junction and adherens junction assembly are being characterized. Calpeptin activates stress fiber assembly in fibroblasts by inhibiting SH2-containing phosphatase-2 (SHP-2), thereby activating Rho-GTPase signaling. Here, we have examined the effects of calpeptin on stress fiber and junctional complex assembly in Madin-Darby canine kidney (MDCK) and LLC-PK epithelial cells. Calpeptin induced disassembly of stress fibers and inhibition of Rho GTPase activity in MDCK cells. Interestingly, calpeptin augmented stress fiber formation in LLC-PK epithelial cells. Calpeptin treatment of MDCK cells resulted in a displacement of zonula occludens-1 (ZO-1) and occludin from cell-cell junctions and a loss of phosphotyrosine on ZO-1 and ZO-2, without any detectable effect on tight junction permeability. Surprisingly, calpeptin increased paracellular permeability in LLC-PK cells even though it did not affect tight junction assembly. Calpeptin also modulated adherens junction assembly in MDCK cells but not in LLC-PK cells. Calpeptin treatment of MDCK cells induced redistribution of E-cadherin and beta-catenin from intercellular junctions and reduced the association of p120ctn with the E-cadherin/catenin complex. Together, our studies demonstrate that calpeptin differentially regulates stress fiber and junctional complex assembly in MDCK and LLC-PK epithelial cells, indicating that these pathways may be regulated in a cell line specific manner. PMID- 12777256 TI - Changing perspectives on ADHD. PMID- 12777257 TI - Maternal lifestyle factors in pregnancy risk of attention deficit hyperactivity disorder and associated behaviors: review of the current evidence. AB - OBJECTIVE: The purpose of this review was to examine the literature assessing the relationship between prenatal exposure to nicotine, alcohol, caffeine, and psychosocial stress during pregnancy to the risk of developing behavioral problems related to attention deficit hyperactivity disorder (ADHD) in childhood. METHOD: PubMed, MEDLINE, EMBASE, and PsycINFO were searched systematically. Studies using DSM diagnostic criteria and other validated diagnostic or screening instruments for ADHD and those examining ADHD symptoms were included. A narrative approach was used because the studies differed too much in methods and data sources to permit a quantitative meta-analysis. RESULTS: Twenty-four studies on nicotine (tobacco smoking), nine on alcohol, one on caffeine, and five on psychosocial stress were identified. All were published between 1973 and 2002. In spite of inconsistencies, the studies on nicotine indicated a greater risk of ADHD-related disorders among children whose mothers smoked during pregnancy. Contradictory findings were reported in the alcohol studies, and no conclusion could be reached on the basis of the caffeine study. Results from studies on psychological stress during pregnancy were inconsistent but indicated a possible modest contribution to ADHD symptoms in the offspring. Many studies suffered from methodological shortcomings, such as recall bias, crude or inaccurate exposure assessments, low statistical power, and lack of or insufficient control of confounders. A general lack of information on familial psychopathology also limited the interpretations. CONCLUSIONS: Exposure to tobacco smoke in utero is suspected to be associated with ADHD and ADHD symptoms in children. Other maternal lifestyle factors during pregnancy may also be associated with these disorders. Further studies are needed to reach conclusions. PMID- 12777259 TI - Schizophrenia, II: amygdalar fiber alteration as etiology? PMID- 12777261 TI - An adolescent with anorexia nervosa and gastrointestinal stromal tumors. PMID- 12777258 TI - Developmental neurocircuitry of motivation in adolescence: a critical period of addiction vulnerability. AB - OBJECTIVE: Epidemiological studies indicate that experimentation with addictive drugs and onset of addictive disorders is primarily concentrated in adolescence and young adulthood. The authors describe basic and clinical data supporting adolescent neurodevelopment as a biologically critical period of greater vulnerability for experimentation with substances and acquisition of substance use disorders. METHOD: The authors reviewed recent literature regarding neurocircuitry underlying motivation, impulsivity, and addiction, with a focus on studies investigating adolescent neurodevelopment. RESULTS: Adolescent neurodevelopment occurs in brain regions associated with motivation, impulsivity, and addiction. Adolescent impulsivity and/or novelty seeking as a transitional trait behavior can be explained in part by maturational changes in frontal cortical and subcortical monoaminergic systems. These developmental processes may advantageously promote learning drives for adaptation to adult roles but may also confer greater vulnerability to the addictive actions of drugs. CONCLUSIONS: An exploration of developmental changes in neurocircuitry involved in impulse control has significant implications for understanding adolescent behavior, addiction vulnerability, and the prevention of addiction in adolescence and adulthood. PMID- 12777263 TI - Neural substrates of decision making in adults with attention deficit hyperactivity disorder. AB - OBJECTIVE: The characteristics of attention deficit hyperactivity disorder (ADHD) include abnormalities in reward responsivity that may interfere with decision making. The study examined reward responsivity in ADHD by comparing the neural correlates of decision making in adults with childhood-onset ADHD and in healthy adults. METHOD: The neural correlates of performance on a decision-making task and a control task were compared in 10 adults with ADHD and 12 age-matched healthy volunteers by using [(15)O]H(2)O positron emission tomography. The decision-making task tested the ability to weigh short-term rewards against long term losses. The control task matched all components of the decision-making task except for the decision-making process and related contingency. RESULTS: The ventral and dorsolateral prefrontal cortex and the insula were activated during performance of the decision-making task in both the ADHD and healthy groups; however, activation in the ADHD group was less extended and did not involve other regions, such as anterior cingulate and hippocampus, that subserve emotion/memory processes. Direct comparison of data from the ADHD subjects and the healthy volunteers suggested that the healthy subjects engaged the hippocampal and insular regions more than did the ADHD subjects and that the ADHD subjects recruited the caudal part of the right anterior cingulate more than did the healthy subjects. CONCLUSIONS: The findings suggest that the neural circuits engaged during decision making differ in subjects with ADHD and healthy comparison subjects. This difference may explain observed deficits in motivated behaviors in ADHD. A better understanding of the nature of these deficits could ultimately be applied to refine treatment strategies for ADHD. PMID- 12777264 TI - National trends in the treatment of attention deficit hyperactivity disorder. AB - OBJECTIVE: Nationally representative descriptive data were presented regarding recent trends in the outpatient treatment of attention deficit hyperactivity disorder (ADHD) in children. METHOD: Service use data were analyzed in children ages 3 to 18 years from two nationally representative surveys of the U.S. general population, the 1987 National Medical Expenditure Survey, and the 1997 Medical Expenditure Panel Survey. Trends in the rates of treatment for ADHD were presented by age, gender, race, family income, and health insurance status. Trends in ADHD treatment were also determined by annual numbers of visits, use of pharmacotherapies, and types of health care professionals. RESULTS: The rate of outpatient treatment for ADHD increased from 0.9 per 100 children in 1987 to 3.4 per 100 children in 1997. Significant increases in the rates of treatment for ADHD were evident across nearly all sociodemographic groups, with the largest increases among children from poor, near-poor, and low-income families and children ages 12 to 18. Among children who received treatment for ADHD, there was a significant decrease in the number of treatment visits but an increase in the number of stimulant prescriptions between 1987 and 1997. CONCLUSIONS: During the decade, there was a marked and broad expansion in access to treatment of children with ADHD but a decline in intensity of treatment, as measured by number of visits. These changes occurred during a period of expanding access to special education services, growth of managed behavioral healthcare, and increased public acceptance of effective psychotropic medications. PMID- 12777266 TI - Attachment and traumatic stress in female holocaust child survivors and their daughters. AB - OBJECTIVE: During the Holocaust, extreme trauma was inflicted on children who experienced it. Two questions were central to the current investigation. First, do survivors of the Holocaust still show marks of their traumatic experiences, even after more than 50 years? Second, was the trauma passed on to the next generation? METHOD: Careful matching of Holocaust survivors and comparison subjects was employed to form a research study design with three generations, including 98 families with a grandmother, a mother, and an infant, who engaged in attachment- and trauma-related interviews, questionnaires, and observational procedures. RESULTS: Holocaust survivors (now grandmothers) showed more signs of traumatic stress and more often lack of resolution of trauma than comparison subjects, but they were not impaired in general adaptation. Also, the traumatic effects did not appear to transmit across generations. CONCLUSIONS: Holocaust survivors may have been able to protect their daughters from their war experiences, although they themselves still suffer from the effects of the Holocaust. PMID- 12777265 TI - Neurobehavioral disinhibition in childhood predicts early age at onset of substance use disorder. AB - OBJECTIVE: This longitudinal study had three aims: 1) determine the extent to which boys at high average risk and low average risk for substance use disorder differ on a construct of neurobehavioral disinhibition, 2) evaluate the capacity of neurobehavioral disinhibition to predict substance use frequency at age 16, and 3) demonstrate the utility of neurobehavioral disinhibition in predicting substance use disorder. METHOD: The authors derived an index of neurobehavioral disinhibition from measures of affect, behavior, and cognition. The neurobehavioral disinhibition score was used to discriminate youth at high and low average risk for substance use disorder and to predict substance use frequency after 4-6 years and substance use disorder after 7-9 years. RESULTS: The neurobehavioral disorder score significantly discriminated boys at high average risk from those at low average risk at ages 10-12. Neurobehavioral disinhibition at age 16, in conjunction with substance use frequency and risk status group, predicted substance use disorder at age 19 with 85% accuracy and accounted for 50% of the variance in Drug Use Screening Inventory overall problem density score. Neurobehavioral disinhibition was a stronger predictor of substance use disorder (odds ratio=6.83) than substance consumption frequency (odds ratio=3.19). CONCLUSIONS: Cross-sectional and longitudinal analyses indicated that neurobehavioral disinhibition is a component of the liability to early age at onset of substance use disorder. PMID- 12777267 TI - Suicide and serious suicide attempts in youth: a multiple-group comparison study. AB - OBJECTIVE: This study compared risk factors for suicide and medically serious nonfatal suicide attempts among youth under 25 years of age. METHOD: Three groups were examined: individuals who committed suicide (N=60), individuals who made serious suicide attempts (N=125), and nonsuicidal community comparison subjects (N=151). Suicidal and nonsuicidal subjects were compared in terms of sociodemographic, childhood, family, psychosocial, and psychiatric factors. RESULTS: Suicides were characterized by male gender, lack of educational qualifications, mood disorder, history of mental health care, and stressful life events. Except for gender, similar risk factors were associated with serious suicide attempts. Suicides and serious suicide attempts were discriminated by gender and mood disorder. Gender differences between suicides and nonfatal suicide attempts were explained by gender differences in methods. CONCLUSIONS: The same risk factors (mood disorder, history of psychiatric care, educational disadvantage, stressful circumstances) play a similar role in suicide and serious suicide attempts. Suicide and suicide attempts are discriminated by mood disorder and gender differences in methods. PMID- 12777268 TI - Altered transcript expression of NMDA receptor-associated postsynaptic proteins in the thalamus of subjects with schizophrenia. AB - OBJECTIVE: NMDA receptor dysfunction has been implicated in the pathophysiology of schizophrenia. The NMDA receptor is a multimeric ligand-gated ion channel, and the obligate NR(1) subunit is expressed as one of eight isoforms due to the alternative splicing of exons 5, 21, and 22. Alternative splicing of NR(1) subunits modulates receptor function by influencing the association of NR(1) with other NMDA receptor subunits and myriad intracellular molecules, such as the postsynaptic density family of proteins that target NMDA receptors to the synaptic membrane and couple it to numerous signal transduction enzymes. Recently, the authors reported that the NMDA receptor subunits NR(1) and NR(2C) are abnormally expressed in the thalamus in schizophrenia. They hypothesized that this reduction is associated with specific NR(1) isoforms and that NMDA receptor related postsynaptic density proteins are abnormally expressed. METHOD: Using in situ hybridization, the authors examined expression of the transcripts encoding NR(1) isoforms containing exons 5, 21, or 22, and the NMDA receptor-related postsynaptic density proteins NF-L, PSD93, PSD95, and SAP102. RESULTS: Reduced NR(1) subunit transcript expression was restricted to exon 22-containing isoforms. Increased expression of the NMDA receptor-associated postsynaptic density proteins NF-L, PSD95, and SAP102 was also detected in the thalamus of subjects with schizophrenia. CONCLUSIONS: These data support the hypothesis of glutamatergic abnormalities in schizophrenia and suggest that glutamatergic dysfunction may occur not only at the level of receptor expression but also within intracellular pathways associated with glutamate receptor-associated signal transduction. PMID- 12777269 TI - Effects of alcohol dependence comorbidity and antipsychotic medication on volumes of the thalamus and pons in schizophrenia. AB - OBJECTIVE: Postmortem and in vivo brain imaging studies have identified abnormalities in the thalamus and the pons in both schizophrenia and alcoholism. The authors sought to determine whether patients with both schizophrenia and alcohol dependence would manifest exaggerated volume deficits in either structure. METHOD: Volumetric measures of the left and right thalamus and the pons were derived from magnetic resonance imaging scans obtained from 27 patients with schizophrenia, 19 patients with schizophrenia and comorbid alcohol dependence, 25 patients with alcohol dependence without comorbid axis I disorders, and 51 healthy comparison subjects. RESULTS: The alcohol-dependent patients had significant volume deficits in both the thalamus and the pons. Among patients with schizophrenia, there were no differences in thalamus volumes between those with and without comorbid alcohol dependence. However, patients with schizophrenia who were taking atypical antipsychotic medications had bilateral thalamic deficits, whereas those taking typical neuroleptics did not. Patients with schizophrenia and comorbid alcohol dependence had deficits in the pons. CONCLUSIONS: Patients with schizophrenia and comorbid alcohol dependence are at risk for alcohol-related reduction of pontine structures that are not necessarily affected by schizophrenia per se. The effect of alcohol dependence on the thalamus in schizophrenic patients may be mitigated by the type of neuroleptic medication they receive. PMID- 12777270 TI - Efficacy of the branched-chain amino acids in the treatment of tardive dyskinesia in men. AB - OBJECTIVE: The efficacy of the branched-chain amino acids in the treatment of tardive dyskinesia in men with psychiatric disorders was tested. METHOD: Public sector psychiatric patients with long histories of antipsychotic treatment and presumably long-standing tardive dyskinesia were randomly assigned to receive branched-chain amino acids or placebo. Treatment frequency was three times a day, 7 days a week for 3 weeks. The efficacy measure was a frequency count of videotaped tardive dyskinesia movements. RESULTS: A robust and highly significant difference was observed between patients who received high-dose branched-chain amino acids (222 mg/kg of body weight t.i.d.) (N=18) and those who received placebo (N=18) in the percent change in tardive dyskinesia symptoms from baseline to the end of the 3-week trial. Significant and marked differences were seen between the two groups at the >/=30% and >/=60% levels of decrease in tardive dyskinesia symptoms. No clinically significant differences were seen between the pre- and posttrial results of physical examinations and laboratory screening tests. Minimal gastrointestinal symptoms occurred during the trial. The reduction in tardive dyskinesia symptoms in the amino acids group was not related to changes in antipsychotic and glucose plasma levels. A mechanism of response related to decreased amine neurotransmitter synthesis was suggested by the significant positive correlations observed between decreases in tardive dyskinesia symptoms and decreases in aromatic amino acid plasma concentrations over the course of the trial. CONCLUSIONS: Branched-chain amino acids constitute a novel, safe treatment for tardive dyskinesia, with a strong potential for providing significant improvement in the diseased physiognomy of the afflicted person. PMID- 12777271 TI - Long-acting injectable risperidone: efficacy and safety of the first long-acting atypical antipsychotic. AB - OBJECTIVE: The authors assessed the efficacy and safety of the first long-acting atypical antipsychotic (long-acting injectable risperidone) in patients with schizophrenia. METHOD: In a 12-week, multicenter, double-blind, randomized study, patients received intramuscular injections every 2 weeks of placebo or long acting risperidone (25 mg, 50 mg, or 75 mg). The primary measure of efficacy was the change in total score on the Positive and Negative Syndrome Scale. RESULTS: Of the 554 patients who were enrolled, 400 entered the double-blind study, and 370 received at least one postbaseline assessment. Mean changes in score of -6.2, -8.5, and -7.4 on the Positive and Negative Syndrome Scale were seen at endpoint for the 25-, 50-, and 75-mg risperidone groups, respectively; all three change scores were significantly different from that seen with placebo (+2.6). Improvements in positive and negative symptoms were also significantly greater in patients receiving risperidone. Long-acting risperidone was well tolerated. Adverse events related to extrapyramidal symptoms were spontaneously reported by 13% of patients receiving placebo and 10% of patients in the 25-mg risperidone group, with higher rates in the 50-mg and 75-mg groups. Severity of extrapyramidal symptoms was mild at baseline and throughout the trial in each treatment group. Mean weight changes were small in the 25-, 50-, and 75-mg risperidone groups (0.5 kg, 1.2 kg, and 1.9 kg, respectively). Injection site pain was rated as low by the patients, consistent with the investigators' pain ratings. CONCLUSIONS: Long-acting injectable risperidone was efficacious and well tolerated and provides both clinicians and patients with a new mode of treatment that can improve the outcome of long-term therapy. PMID- 12777272 TI - Effect size of symptom status in withdrawal of typical antipsychotics and subsequent clozapine treatment in patients with treatment-resistant schizophrenia. AB - OBJECTIVE: In light of the efficacy of newer antipsychotic agents and the possibility that drug withdrawal may negatively affect subsequent drug response, concern has arisen that the use of placebo in schizophrenia research may be unethical. This study examines the effect size of symptom exacerbation during drug washout with placebo and the effects of drug washout on the efficacy of subsequent drug treatment. METHOD: Fifty patients with treatment-resistant schizophrenia hospitalized on a research unit participated in a double-blind longitudinal study of the effects of drug washout after chronic treatment with a typical antipsychotic and before prospective treatment with clozapine. Brief Psychiatric Rating Scale (BPRS) scores were analyzed to examine drug effects and effect sizes for baseline treatment with a typical antipsychotic (>6 months treatment), drug washout with placebo (mean=34 days), early treatment with clozapine (mean=42 days, mean dose=345.0 mg/day), and optimal clozapine treatment (mean=83 days, mean dose=450.5 mg/day). RESULTS: Patients' BPRS total, positive, and negative symptom scores significantly increased during placebo washout, compared with baseline treatment, and significantly decreased with administration of clozapine, compared with placebo washout and baseline treatment. However, 30% of patients showed some symptom improvement during placebo washout. The effect sizes for the BPRS total score were 0.63 for baseline treatment versus placebo washout, 1.10 for optimal clozapine treatment versus placebo washout, and 0.82 for optimal clozapine treatment versus baseline treatment. CONCLUSIONS: Symptom exacerbation induced by drug withdrawal in patients with treatment-resistant schizophrenia did not impede subsequent responsiveness to clozapine. The effect size for clozapine, compared with typical antipsychotics, suggests that the drug washout longitudinal design is useful for establishing a drug-free baseline and for investigating drug response, while requiring relatively few subjects. PMID- 12777273 TI - Assessment of four stakeholder groups' preferences concerning outpatient commitment for persons with schizophrenia. AB - OBJECTIVE: Study findings indicating that involuntary outpatient commitment can improve treatment outcomes among persons with severe mental illness remain controversial. Opponents of outpatient commitment argue that its coerciveness is unacceptable even given its arguable benefits. However, it is unclear to what extent the public debate surrounding outpatient commitment represents the preferences of persons with a stake in the benefit or harm resulting from outpatient commitment. This study examines and compares views of outpatient commitment among four stakeholder groups: 1) persons in treatment for schizophrenia and related disorders, 2) family members of persons with these disorders, 3) clinicians treating persons with these disorders, and 4) members of the general public. METHOD: Subjects from the Piedmont region of North Carolina who were members of the four stakeholder groups were presented with short vignettes that depicted potential outcomes that were associated alternatively with outpatient commitment and with voluntary treatment. Subjects rated each vignette according to how positively or negatively they viewed the overall situation for the individual described. Multivariate regression techniques were used to estimate preference weights for each stakeholder group. RESULTS: With some exceptions, each group gave the highest preference to avoiding involuntary hospitalization, followed by avoiding interpersonal violence and maintaining good interpersonal relationships. No group gave appreciable importance to outpatient commitment, which suggests that avoiding its coerciveness is a lesser concern compared to other outcomes. CONCLUSIONS: The findings suggest that these stakeholders are willing to accept the coerciveness of outpatient commitment to gain improved outcomes for certain persons with schizophrenia and related disorders. PMID- 12777274 TI - Risk factors for depression among elderly community subjects: a systematic review and meta-analysis. AB - OBJECTIVE: The goal of this study was to determine risk factors for depression among elderly community subjects. METHOD: MEDLINE and PsycINFO were searched for potentially relevant articles published from January 1966 to June 2001 and from January 1967 to June 2001, respectively. The bibliographies of relevant articles were searched for additional references. Twenty studies met the following six inclusion criteria: original research reported in an English or French publication, study group of community residents, age of subjects 50 years or more, prospective study design, examination of at least one risk factor, and use of an acceptable definition of depression. The validity of studies was assessed according to the four primary criteria for risk factor studies described by the Evidence-Based Medicine Working Group. Information about group size at baseline and follow-up, age, proportion of men, depression criteria, exclusion criteria at baseline, length of follow-up, number of incident cases of depression, and risk factors was abstracted from each report. RESULTS: Follow-up of the inception cohort was incomplete in most studies. In the qualitative meta-analysis, risk factors identified by both univariate and multivariate techniques in at least two studies each were disability, new medical illness, poor health status, prior depression, poor self-perceived health, and bereavement. In the quantitative meta analysis, bereavement, sleep disturbance, disability, prior depression, and female gender were significant risk factors. CONCLUSIONS: Despite the methodologic limitations of the studies and this meta-analysis, bereavement, sleep disturbance, disability, prior depression, and female gender appear to be important risk factors for depression among elderly community subjects. PMID- 12777275 TI - Does cognitive recovery after treatment of poststroke depression last? A 2-year follow-up of cognitive function associated with poststroke depression. AB - OBJECTIVE: Cognitive impairment is common after stroke and may be caused by poststroke depression. Remission of poststroke major depression after treatment has been associated with improvement in cognitive function. The current study was designed to examine how long that cognitive improvement lasts and to compare depressed patients' cognitive status with that of nondepressed patients with comparable lesions. METHOD: Seventeen patients with poststroke depression and cognitive impairment who had early and sustained remission of their depression during a double-blind treatment study were compared with 42 nondepressed stroke patients who remained nondepressed throughout the follow-up. Mood and cognitive function were followed-up over 2 years with the Hamilton Depression Rating Scale and Mini-Mental State Examination (MMSE). RESULTS: In the patients with early and sustained remission of depression, there was rapid improvement of cognitive function, which was maintained over 2 years. Their initial MMSE score of 23.3 (SD=4.2) improved to 26.6 (SD=3.5) at 3 months and was 26.1 (SD=3.6) at 2 years. The nondepressed patients showed essentially no change in cognitive function over 2 years (initial MMSE score: mean=26.3, SD=3.1; score at 2-year follow-up: mean=25.7, SD=4.1). CONCLUSIONS: Cognitive function, once improved after remission of poststroke depression, is likely to remain stable over the next 2 years in the absence of subsequent reinjury to the central nervous system. Cognitive impairment due to poststroke depression is reversible and can be quantified separately from cognitive impairment on the basis of the location and extent of ischemic brain damage. PMID- 12777276 TI - Patients who strive to be ill: factitious disorder with physical symptoms. AB - OBJECTIVE: Factitious disorder with physical symptoms characterizes patients who strive to appear medically ill and assume the sick role. Clinical suspicion is highest for female health care workers in the fourth decade of life. This study was designed to analyze the diagnosis of factitious disorder, the demographics of affected patients, and intervention and treatment. METHOD: Retrospective examination was of 93 patients diagnosed during 21 years. Two raters agreed on subject eligibility on the basis of DSM-IV criteria and absence of a somatoform disorder and a plausible medical explanation. RESULTS: The group included 67 women (72.0%); mean age was 30.7 years (SD=8.0) for women and 40.0 years (SD=13.3) for men. Mean age at onset was 25.0 years (SD=7.4). Health care training or jobs were more common for women (65.7%) than men (11.5%). Most often, inexplicable laboratory results established the diagnosis. Eighty had psychiatric consultations; 71 were confronted about their role in the illness. Only 16 acknowledged factitious behavior. Follow-up data were available for only 28 patients (30.1%); maximum duration of follow-up was 156 months. Two patients were known to have died. Few patients pursued psychiatric treatment. Eighteen left the hospital against medical advice. CONCLUSIONS: Factitious disorder affects men and women with different demographic profiles. Diagnosis must be based on careful examination of behavior, motivation, and medical history and not on a stereotype. Laboratory data and outside medical records help identify suspicious circumstances and inconsistencies. Confrontation does not appear to lead to patient acknowledgment and should not be considered necessary for management. PMID- 12777277 TI - Modification of DSM-IV criteria for depressed preschool children. AB - OBJECTIVE: This study compared the severity of depression in preschoolers diagnosed by standard versus modified DSM-IV criteria for major depression. METHOD: A group of 145 preschoolers and their caregivers underwent a diagnostic assessment for preschool children. A factor analysis of depressive symptoms from the group was performed to derive a depression severity score. Scores were compared among four groups: standard DSM-IV major depression, modified DSM-IV major depression, DSM-IV attention deficit hyperactivity disorder and/or oppositional defiant disorder, and no disorder. RESULTS: A hierarchy in severity emerged, with significant differences among all four groups. Preschoolers meeting standard criteria displayed the highest severity, followed by those who met modified criteria. Both depressed groups had significantly higher severity than the two comparison groups. CONCLUSIONS: Standard DSM-IV criteria captured the most severely affected preschoolers, missing a substantial proportion of children with potentially clinically significant but less severe symptoms who were captured by modified DSM-IV criteria. PMID- 12777278 TI - Facial expression recognition in adolescents with mood and anxiety disorders. AB - OBJECTIVE: The authors examined facial expression recognition in adolescents with mood and anxiety disorders. METHOD: Standard facial emotion identification tests were given to youth with bipolar disorder (N=11) or DSM-IV anxiety disorders (N=10) and a group of healthy comparison subjects (N=25). RESULTS: Relative to the anxiety disorder and healthy comparison groups, the subjects with bipolar disorder made more emotion recognition errors when presented with faces of children. Unlike the anxious and comparison subjects, bipolar disorder youth were prone to misidentify faces as angry. No differences in emotion recognition errors were seen when the adolescents were presented with adult faces. CONCLUSIONS: A bias to misinterpret the facial expressions of peers as angry may characterize youth with bipolar disorder but not youth with anxiety disorders. This bias may relate to social impairment in youth with bipolar disorder. PMID- 12777279 TI - Habit reversal versus supportive psychotherapy for Tourette's disorder: a randomized controlled trial. AB - OBJECTIVE: The authors investigated the efficacy of habit reversal for Tourette's disorder, which is characterized by multiple motor and vocal tics. METHOD: Thirty two patients with Tourette's disorder were randomly assigned to 14 sessions of either habit reversal or supportive psychotherapy. Habit reversal consisted of awareness training, self-monitoring, relaxation training, competing response training, and contingency management. Changes in severity of Tourette's disorder and psychosocial impairment were investigated over the course of the 14-session treatment for the 29 patients who completed at least eight treatment sessions. RESULTS: In contrast to the 13 patients in the supportive psychotherapy group, the 16 patients in the habit reversal group improved significantly. The habit reversal patients remained significantly improved over pretreatment at 10-month follow-up. CONCLUSIONS: Habit reversal may be an effective behavioral treatment for Tourette's disorder. PMID- 12777280 TI - Low medial and lateral right pulvinar volumes in schizophrenia: a postmortem study. AB - OBJECTIVE: In this study, the volume and neuronal number of the pulvinar thalamic nucleus in schizophrenia patients were measured. METHOD: The authors examined medial and lateral pulvinar nuclei bilaterally in 27 patients with schizophrenia and 28 normal comparison subjects. RESULTS: In the comparison subjects, the medial pulvinar was larger on the right. The right but not left pulvinar nuclei were smaller in the schizophrenia patients than in the comparison subjects. The number of neurons showed similar trends. CONCLUSIONS: These findings confirm low pulvinar volume in schizophrenia and show that it affects both medial and lateral nuclei. The lateralized findings may reflect pulvinar connections with asymmetrical neocortical regions and their asymmetrical involvement in schizophrenia. PMID- 12777281 TI - Effects of alcoholism and gender on brain metabolism. AB - OBJECTIVE: Proton magnetic resonance spectroscopy was used to evaluate gender influences on alcohol-associated changes in brain metabolism. METHOD: Concentrations of N-acetylaspartate, choline-containing compounds, myo-inositol, and creatine plus phosphocreatine in frontal lobe gray matter and white matter were estimated in eight women and 17 men who were recently detoxified from long term alcoholism. Twelve women and 13 men with no history of alcoholism were used as a comparison group. RESULTS: In male and female alcoholics, frontal lobe white matter concentrations of N-acetylaspartate were significantly lower (-8.8%) than those seen in nonalcoholic comparison subjects. In the frontal lobe gray matter region, a significant alcoholism status-by-gender interaction and follow-up analyses revealed that female alcoholics had significantly lower N acetylaspartate concentrations (-10.73%) relative to female comparison subjects, while male alcoholics and male comparison subjects had similar levels of this metabolite (<1% difference). CONCLUSIONS: Lower concentrations of white matter N acetylaspartate, which may indicate neuronal loss or dysfunction, is equally severe in men and women with comparable alcohol abuse histories. However, female alcoholics exhibited significantly less N-acetylaspartate in frontal gray matter relative to female nonalcoholic comparison subjects, which could mean that female alcoholics are more susceptible to gray matter injury than their male counterparts. However, this finding could also be explained by higher-than expected levels of N-acetylaspartate in the healthy female comparison group. PMID- 12777282 TI - Childhood central nervous system viral infections and adult schizophrenia. AB - OBJECTIVE: An earlier Finnish cohort study suggested that childhood viral CNS infections are associated with a fivefold increased odds of developing schizophrenia in adulthood. The authors sought to replicate this finding. METHOD: From the archives of the Department of Virology of the National Public Health Institute in Finland, 320 individuals born between 1960 and 1976 who had suffered virologically confirmed CNS infections before their 15th birthdays were identified. Of the infections, 202 had been caused by enteroviruses. The sample was followed up in the 1969-2000 records of the National Hospital Discharge Register of Finland to identify all cases of schizophrenia that emerged. RESULTS: The cumulative incidence of schizophrenia was 0.94% in the whole sample and 0.99% among individuals who had suffered enteroviral infections. These rates are comparable to that found in the general population. CONCLUSIONS: Childhood viral CNS infections were not associated with increased risk of schizophrenia. PMID- 12777283 TI - Fetal hypoxia, genetic risk, and schizophrenia. PMID- 12777284 TI - Comparing depression treatments. PMID- 12777288 TI - Childhood trauma and depression. PMID- 12777289 TI - Randomized controlled trials. PMID- 12777290 TI - Mental disorders among military personnel. PMID- 12777291 TI - Mental disorders among military personnel. PMID- 12777294 TI - Suicide and major depression. PMID- 12777295 TI - Sensitivity of the D8/17 assay. PMID- 12777313 TI - Role for chymase in heart failure: angiotensin II-dependent or -independent mechanisms? PMID- 12777314 TI - Aldosterone blockade in patients with acute myocardial infarction. PMID- 12777315 TI - Coronary artery calcium and cardiac events: is electron-beam tomography ready for prime time? PMID- 12777316 TI - Less heart is more. PMID- 12777317 TI - Fibrinolysis for acute myocardial infarction: the future is here and now. PMID- 12777318 TI - Fibrinolytic therapy: is it a treatment of the past? PMID- 12777319 TI - Editor's commentary: One size does not fit all. PMID- 12777320 TI - Images in cardiovascular medicine. Anomalous coronary arteries by electron beam angiography. PMID- 12777321 TI - Cardiology patient page. Bacterial endocarditis: the disease, treatment, and prevention. PMID- 12777322 TI - Images in cardiovascular medicine. Cardiac sarcoidosis evaluated by delayed enhanced magnetic resonance imaging. PMID- 12777323 TI - Unraveling Reaven's syndrome X: serum insulin-like growth factor-I and cardiovascular disease. PMID- 12777324 TI - Low serum insulin-like growth factor I is associated with increased risk of ischemic heart disease. PMID- 12777325 TI - Regional pathologies and globalization of clinical trials: has the time for regional trials arrived? PMID- 12777326 TI - Stem cell-derived cardiomyocytes demonstrate arrhythmic potential. PMID- 12777327 TI - Left ventricular remodeling after primary coronary angioplasty: patterns of left ventricular dilation and long-term prognostic implications. PMID- 12777328 TI - Long-term use of contraceptive depot medroxyprogesterone acetate in young women impairs arterial endothelial function assessed by cardiovascular magnetic resonance. PMID- 12777329 TI - Management of vasovagal syncope: controlling or aborting faints by leg crossing and muscle tensing. PMID- 12777330 TI - Low-density lipoprotein, non-high-density lipoprotein, and apolipoprotein B as targets of lipid-lowering therapy. PMID- 12777331 TI - Incremental benefits seen for electron-beam tomography. PMID- 12777333 TI - Work and employment for people with psychiatric disabilities. PMID- 12777334 TI - Taking another tilt at high secure hospitals. The Tilt Report and its consequences for secure psychiatric services. PMID- 12777335 TI - Psychiatry and the human sciences. PMID- 12777336 TI - Suicide terrorism: a case of folie a plusieurs? PMID- 12777337 TI - Is cognitive-behavioural therapy a worthwhile treatment for psychosis? PMID- 12777338 TI - Repetitive transcranial magnetic stimulation for the treatment of depression. Systematic review and meta-analysis. AB - BACKGROUND: Repetitive transcranial magnetic stimulation (rTMS) may be useful in the treatment of depression but results from trials have been inconclusive to date. AIMS: To assess the efficacy of rTMS in treating depression. METHOD: We conducted a systematic review of randomised controlled trials that compared rTMS with sham in patients with depression. We assessed the quality of design of all studies and conducted a meta-analysis of data from trials with similar rTMS delivery. RESULTS: We included a total of 14 trials. The quality of the included studies was low. Pooled analysis using the Hamilton Rating Scale for Depression showed an effect in favour of rTMS compared with sham after 2 weeks of treatment (standardised mean difference=-0.35; 95% CI -0.66 to -0.04), but this was not significant at the 2-week follow-up (standardised mean difference=-0.33; 95% CI 0.84 to 0.17). CONCLUSIONS: Current trials are of low quality and provide insufficient evidence to support the use of rTMS in the treatment of depression. PMID- 12777339 TI - Older community residents with depression: long-term treatment with sertraline. Randomised, double-blind, placebo-controlled study. AB - BACKGROUND: Despite a growing use of selective serotonin reuptake inhibitors in older people, only one trial has examined their prophylactic efficacy in people aged 65 years and over. AIMS: To examine the efficacy of sertraline in preventing the recurrence of depression in older people living in the community. METHOD: Participants were openly treated with sertraline and then randomised into a double-blind, placebo-controlled continuation/maintenance study of about 2 years duration. Drug dosage was maintained at levels that achieved remission. RESULTS: No significant difference between the sertraline and placebo groups was found in the proportion of recurrences (-7.9%; 95% CI -28.06 to 12.23). Increased age and minor residual symptoms during the continuation phase were associated with recurrence. CONCLUSIONS: Sertraline at therapeutic dosage does not provide significant protection against recurrence. PMID- 12777340 TI - Tapering off long-term benzodiazepine use with or without group cognitive behavioural therapy: three-condition, randomised controlled trial. AB - BACKGROUND: Benzodiazepine withdrawal programmes have never been experimentally compared with a nonintervention control condition. AIMS: To evaluate the efficacy and feasibility of tapering off long-term benzodiazepine use in general practice, and to evaluate the value of additional group cognitive-behavioural therapy (CBT). METHOD: A 3-month randomised, 3-month controlled trial was conducted in which 180 people attempting to discontinue long-term benzodiazepine use were assigned to tapering off plus group CBT, tapering off alone or usual care. RESULTS: Tapering off led to a significantly higher proportion of successful discontinuations than usual care (62% nu. 21%). Adding group CBT did not increase the success rate (58% v. 62%). Neither successful discontinuation nor intervention type affected psychological functioning. Both tapering strategies showed good feasibilityin general practice. CONCLUSIONS: Tapering off is a feasible and effective way of discontinuing long-term benzodiazepine use in general practice. The addition of group CBT is of limited value. PMID- 12777341 TI - Modelling the impact of clozapine on suicide in patients with treatment-resistant schizophrenia in the UK. AB - BACKGROUND: Schizophrenia is a major cause of suicide, and symptoms characteristic of treatment-resistant disease are strong risk factors. Clozapine reduces symptoms in 60% of such patients and significantly decreases the risk of suicide. AIMS: To model the impact of increased clozapine prescribing on lives saved and resource utilisation. METHOD: A model was built to compare current levels of clozapine prescribing with a scenario in which all suitable patients with treatment-resistant schizophrenia received clozapine. RESULTS: It was estimated that an average of 53 lives could be saved in the UK each year. If clozapine is cost-neutral, the cost per life-year saved is pound 5108. If clozapine achieves a 10% reduction in annual support costs, the net saving is pound 8.7 million per annum. An average of 167 acute beds would be freed each year. CONCLUSIONS: The use of clozapine in treatment-resistant schizophrenia saves lives, frees resources and is cost-effective. PMID- 12777342 TI - Psychiatric morbidity and substance use in young people aged 13-15 years: results from the Child and Adolescent Survey of Mental Health. AB - BACKGROUND: Psychoactive substance use is strongly associated with psychiatric morbidity in both adults and adolescents. AIMS: To determine which of alcohol, nicotine and cannabisis mostclosely linked to psychiatric disorders in early adolescence. METHOD: Data from 2624 adolescents aged 13-15 years were drawn from a national mental health survey of children. The relationship between psychiatric morbidity and smoking, drinking and cannabis use was examined by logistic regression analyses. RESULTS: Having a psychiatric disorder was associated with an increased risk of substance use. Greater involvement with any one substance increased the risk of other substance use. Analyses of the interactions between smoking, drinking and cannabis use indicated that the relationship between substance use and psychiatric morbidity was primarily explained by regular smoking and (to a lesser extent) regular cannabis use. CONCLUSIONS: In this sample, links between substance use and psychiatric disorders were primarily accounted for by smoking. The strong relationship is likely to be due to a combination of underlying individual constitutional factors and drug-specific effects resulting from consumption over the period of adolescent development and growth. PMID- 12777343 TI - Morphology of the anterior cingulate cortex in young men at ultra-high risk of developing a psychotic illness. AB - BACKGROUND: The anterior cingulate cortex (ACC) is consistently implicated in the pathophysiology of schizophrenia, and our own work has identified morphological anomalies in the ACC of people with this disorder. AIMS: To examine whether ACC morphological anomalies are present in a group at ultra-high risk of psychosis and whether such anomalies can be used to predict the subsequent development of a psychotic illness. METHOD: Magnetic resonance imaging of 75 healthy volunteers and 63 people at ultra-high risk of developing a psychotic disorder (all right handed males) was used to examine ACC sulcal and gyral features. RESULTS: Compared with the controls, significantly fewer people in the ultra-high risk group had a well-developed left paracingulate sulcus and significantly more had an interrupted left cingulate sulcus. There was no difference between those who did (n=21) and did not (n=42) subsequently develop a psychotic illness. CONCLUSIONS: Although ACC anomalies are present in young people considered to be at ultra-high risk of psychosis, they do not identify individuals who subsequently make the transition to psychosis. PMID- 12777344 TI - Engagement of brain areas implicated in processing inner speech in people with auditory hallucinations. AB - BACKGROUND: The neurocognitive basis of auditory hallucinations is unclear, but there is increasing evidence implicating abnormalities in processing inner speech. Previous studies have shown that people with schizophrenia who were prone to auditory hallucinations demonstrated attenuated activation of brain areas during the monitoring of inner speech. AIMS: To investigate whether the same pattern of functional abnormalities would be evident as the rate of inner speech production was varied. METHOD: Eight people with schizophrenia who had a history of prominent auditory hallucinations and eight control participants were studied using functional magnetic resonance imaging while the rate of inner speech generation was varied experimentally. RESULTS: When the rate of inner speech generation was increased, the participants with schizophrenia showed a relatively attenuated response in the right temporal, parietal, parahippocampal and cerebellar cortex. CONCLUSIONS: In people with schizophrenia who are prone to auditory hallucinations, increasing the demands on the processing of inner speech is associated with attenuated engagement of the brain areas implicated in verbal self-monitoring. PMID- 12777345 TI - The Aberfan disaster: 33-year follow-up of survivors. AB - BACKGROUND: Experiencing life-threatening events often contributes to the onset of such psychiatric conditions as post-traumatic stress disorder (PTSD). Children can develop PTSD; however, there is controversy over whether PTSD symptoms decrease or persist over time. AIMS: To examine the long-term effects of surviving the 1966 Aberfan disaster in childhood. METHOD: Survivors (n=41) were compared with controls (n=72) matched for age and background. All were interviewed using the Composite International Diagnostic Interview, measures of current health and social satisfaction, and the General Health Questionnaire. The survivor group also completed the Impact of Event Scale to assess current levels of PTSD. RESULTS: Nineteen (46%; 95% CI 31-61) survivors had had PTSD at some point since the disaster, compared with 12 (20%; 95% CI 10-30) controls (OR=3.38 (95% CI 1.40-8.47)). Of the survivors,12 (29%; 95% CI 15-43) met diagnostic criteria for current PTSD. Survivors were not at a significantly increased risk of anxiety, depression or substance misuse. CONCLUSIONS: Trauma in childhood can lead to PTSD, and PTSD symptoms can persist for as long as 33 years into adult life. Rates of other psychopathological disorders are not necessarily raised after life-threatening childhood trauma. PMID- 12777346 TI - Suicide following deliberate self-harm: long-term follow-up of patients who presented to a general hospital. AB - BACKGROUND: Deliberate self-harm (DSH) is the strongest risk factor for future suicide. Up-to-date information on the extent of risk is lacking. AIMS: To investigate the risk of suicide after DSH during a long follow-up period. METHOD: A mortality follow-up study to 2000 was conducted on 11 583 patients who presented to hospital after DSH between 1978 and 1997. Data were obtained from a general hospital DSH register in Oxford and the Office for National Statistics, and from equivalent mortality registers in Scotland and Northern Ireland. RESULTS: Three hundred patients had died by suicide or probable suicide. The risk in the first year of follow-up was 0.7% (95% CI 0.6-0.9%), which was 66 (95% CI 52-82) times the annual risk of suicide in the general population. The risk after 5 years was 1.7%, at 10 years 2.4% and at 15 years 3.0%. The risk was far higher in men than in women (hazard ratio 2.8,95% CI 2.2-3.6). In both genders it increased markedly with age at initial presentation. CONCLUSIONS: Following DSH there is a significant and persistent risk of suicide, which varies markedly between genders and age groups. Reduction in the risk of suicide following DSH must be a key element in national suicide prevention strategies. PMID- 12777347 TI - Childhood trauma and hallucinations in bipolar affective disorder: preliminary investigation. AB - BACKGROUND: Strong evidence exists for an association between childhood trauma, particularly childhood sexual abuse, and hallucinations in schizophrenia. Hallucinations are also well-documented symptoms in people with bipolar affective disorder. AIMS: To investigate the relationship between childhood sexual abuse and other childhood traumas and hallucinations in people with bipolar affective disorder. METHOD: A sample of 96 participants was drawn from the Medical Research Council multi-centre trial of cognitive-behavioural therapy for bipolar affective disorder. The trial therapists recorded spontaneous reports of childhood sexual abuse made during the course of therapy. Symptom data were collected by trained research assistants masked to the hypothesis. RESULTS: A significant association was found between those reporting general trauma (n=38) and auditory hallucinations. A highly significant association was found between those reporting childhood sexual abuse (n=15) and auditory hallucinations. CONCLUSIONS: The relationship between childhood sexual abuse and hallucinations in bipolar disorder warrants further investigation. PMID- 12777348 TI - How should advance statements be implemented? PMID- 12777349 TI - High-security hospitals. PMID- 12777350 TI - Rivastigmine and QT interval prolongation. PMID- 12777351 TI - The antidepressant debate should move on. PMID- 12777352 TI - The antidepressant debate should move on. PMID- 12777353 TI - Preserve psychoanalysis from too much neuroscience. PMID- 12777354 TI - Talking about cognitive analytic therapy. PMID- 12777356 TI - Adjusting for screening history in epidemiologic studies of cancer: why, when, and how to do it. AB - In epidemiologic studies of cancer, differences between exposed and nonexposed persons with regard to a history of cancer screening during the time the malignancy (or an antecedent lesion) typically is present prior to diagnosis can be a source of confounding if one of the following conditions is present: (1) the screening modality identifies premalignant changes whose treatment has the potential to prevent the cancer from developing; or (2) the number of cases included in the study would have been smaller but for the presence of screening. These situations occur commonly, arguing that consideration be given to screening history in the design (with attention to distinguishing true screening tests from those administered to persons with signs or symptoms of cancer) and analysis of epidemiologic studies of those cancers for which screening modalities are in use in the study population. PMID- 12777357 TI - Invited commentary: screening as a nuisance variable in cancer epidemiology: methodological considerations. PMID- 12777358 TI - Weiss responds to "Screening as a nuisance variable in cancer epidemiology": for which aspects of a study subject's screening history should we control? PMID- 12777359 TI - Cancer in a population-based cohort of men and women in registered homosexual partnerships. AB - Cancer patterns among broad populations of homosexual men and women have not been studied systematically. The authors followed 1,614 women and 3,391 men in Denmark for cancer from their first registration for marriage-like homosexual partnership between 1989 and 1997. Ratios of observed to expected cancers measured relative risk. Women in homosexual partnerships had cancer risks similar to those of Danish women in general (overall relative risk (RR) = 0.9, 95% confidence interval (CI): 0.6, 1.4), but only one woman developed cervical carcinoma in situ versus 5.8 women expected (RR = 0.2, 95% CI: 0.0, 0.97). Overall, men in homosexual partnerships were at elevated cancer risk (RR = 2.1, 95% CI: 1.8, 2.5), due mainly to human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS)-associated Kaposi's sarcoma (RR = 136, 95% CI: 96, 186) and non Hodgkin's lymphoma (RR = 15.1, 95% CI: 10.4, 21.4). Anal squamous carcinoma also occurred in excess (RR = 31.2, 95% CI: 8.4, 79.8). After exclusion of Kaposi's sarcoma, non-Hodgkin's lymphoma, and anal squamous carcinoma, no unusual cancer risk remained (RR = 1.0, 95% CI: 0.8, 1.3). With anal squamous carcinoma and HIV/AIDS-associated cancers as notable exceptions in men, cancer incidence rates among homosexual persons in marriage-like partnerships are similar to those prevailing in society at large. PMID- 12777360 TI - Changes in smoking status affect women more than men: results of the Lung Health Study. AB - Lung Health Study participants were smokers aged 35-60 years with mild lung function impairment who participated in a 5-year, 10-center (nine in the United States, one in Canada) clinical trial in 1986-1994. The authors compared the relation of randomized treatment assignments and of smoking history during the study with changes in lung function between men and women. Spirometry was performed annually, and 3,348 men and 1,998 women attended the follow-up clinic visit that included spirometry at year 5. This paper reports on an analysis of changes in lung function by gender, treatment group, and three smoking history categories: sustained quitters, intermittent quitters, and continuing smokers. Among participants who quit smoking in the first year, mean forced expiratory volume in 1 second (FEV(1)) expressed as a percentage of the predicted value of FEV(1 )given the person's age, height, gender, and race (FEV(1)%) increased more in women (3.7% of predicted) than in men (1.6% of predicted) (p < 0.001). Across the 5-year follow-up period, among sustained quitters, women gained more in FEV(1)% of predicted than did men. Methacholine reactivity was more strongly related to rates of decline in women than in men (p < 0.001). Therefore, among persons at risk for chronic obstructive pulmonary disease, smoking cessation has an even clearer advantage for women than it does for men. PMID- 12777361 TI - Diet Quality Index as a predictor of short-term mortality in the American Cancer Society Cancer Prevention Study II Nutrition Cohort. AB - The Diet Quality Index (DQI) was developed to measure overall dietary patterns and to predict chronic disease risk. This study examined associations between DQI and short-term all-cause, all-circulatory-disease, and all-cancer mortality in the American Cancer Society Cancer Prevention Study II Nutrition Cohort, a cohort of US adults aged 50-79 years enrolled in a prospective study. After 4 years of follow-up (1992-1996), there were 869 deaths among 63,109 women and 1,736 deaths among 52,724 men. All study participants reported being disease free at baseline in 1992-1993. In age-adjusted Cox models, a higher DQI, which was indicative of a poorer quality diet, was positively related to all-cause and all-circulatory disease mortality rates in both women and men and to cancer mortality in men only. However, in fully adjusted Cox models, only circulatory disease mortality was clearly positively related to DQI and only in women (medium-low-quality diet vs. highest-quality diet: rate ratio = 1.86, 95% confidence interval: 1.19, 2.89). Although trend tests indicated significant positive relations between DQI and all-cause mortality, effects were small (rate ratios /=25 years who were exposed to job strain for 50% of their work life was 4.8 mmHg (95% confidence interval: -3.7, 13.4) higher at work and 7.9 mmHg (95% confidence interval: 0.8, 15.0) higher at home than that of men with no past exposure, independent of current exposure. Evidence was inconsistent for the hypothesis of rapid induction of/recovery from the effects of job strain on blood pressure, and there was little effect of past job strain on diastolic blood pressure. These findings provide some support for the hypothesis of an effect of cumulative burden of exposure to job strain on systolic blood pressure. PMID- 12777364 TI - Dietary intakes of fat and risk of Parkinson's disease. AB - Previous epidemiologic studies have generated inconsistent results regarding the associations between fat intakes and risk of Parkinson's disease. The authors investigated these associations in two large, prospective US cohorts. They documented 191 incident cases of Parkinson's disease in men (1986-1998) and 168 in women (1980-1998) during the follow-up. Overall, intakes of total fat or major types of fat were not significantly associated with the risk. The relative risks comparing the highest quintile of animal fat intake with the lowest were 1.42 for men (95% confidence interval (CI): 0.91, 2.20; p for trend = 0.1) and 0.65 for women (95% CI: 0.36, 1.16; p for trend = 0.3). For men, but not women, replacement of polyunsaturated fat with saturated fat was associated with a significantly increased risk (5% of energy intake, relative risk (RR) = 1.83, 95% CI: 1.10, 3.03). Of the individual polyunsaturated fatty acids, arachidonic acid tended to be inversely associated with the risk (pooled RR between extreme quintiles = 0.65, 95% CI: 0.46, 0.91; p for trend = 0.05). Results do not support an important role of overall fat intake in the pathogenesis of Parkinson's disease, but a possible adverse effect of saturated fat for men could not be excluded. PMID- 12777365 TI - Incidence of Parkinson's disease: variation by age, gender, and race/ethnicity. AB - The goal of this study was to estimate the incidence of Parkinson's disease by age, gender, and ethnicity. Newly diagnosed Parkinson's disease cases in 1994 1995 were identified among members of the Kaiser Permanente Medical Care Program of Northern California, a large health maintenance organization. Each case met modified standardized criteria/Hughes diagnostic criteria as applied by a movement disorder specialist. Incidence rates per 100,000 person-years were calculated using the Kaiser Permanente membership information as the denominator and adjusted for age and/or gender using the direct method of standardization. A total of 588 newly diagnosed (incident) cases of Parkinson's disease were identified, which gave an overall annualized age- and gender-adjusted incidence rate of 13.4 per 100,000 (95% confidence interval (CI): 11.4, 15.5). The incidence rapidly increased over the age of 60 years, with only 4% of the cases being under the age of 50 years. The rate for men (19.0 per 100,000, 95% CI: 16.1, 21.8) was 91% higher than that for women (9.9 per 100,000, 95% CI: 7.6, 12.2). The age- and gender-adjusted rate per 100,000 was highest among Hispanics (16.6, 95% CI: 12.0, 21.3), followed by non-Hispanic Whites (13.6, 95% CI: 11.5, 15.7), Asians (11.3, 95% CI: 7.2, 15.3), and Blacks (10.2, 95% CI: 6.4, 14.0). These data suggest that the incidence of Parkinson's disease varies by race/ethnicity. PMID- 12777366 TI - Changes in functional status attributable to hip fracture: a comparison of hip fracture patients to community-dwelling aged. AB - Disability attributable to hip fracture regarding activities of daily living was evaluated by comparing 594 hip fracture patients entering eight hospitals in Baltimore, Maryland, in 1990-1991 with community-dwelling aged from the Established Populations for Epidemiologic Studies of the Elderly (EPESE) cohort matched on age, sex, and walking ability. Subjects were assessed at baseline (prefracture report for patients), 12 months, and 24 months. At baseline, 26% of both groups had walking disability, 12-14% had transferring disability, and 6-8% evidenced grooming disability. At 12 and 24 months, about 50% of hip fracture patients were walking disabled compared with 21-29% of EPESE respondents after the authors controlled for age, sex, comorbidities, and functional status (excess disability attributable to hip fracture, i.e., attributable disability, of 26 additional cases of disability per 100 persons in the hip fracture cohort during follow-up). Likewise, hip fracture patients experienced more disability regarding transferring (38-39% vs. 10-18%; attributable disability, approximately 22 cases per 100 persons) and grooming (17-19% vs. 7-15%; attributable disability, approximately six cases per 100 persons). Thus, results showed that hip fracture patients had substantially more activities of daily living disability than that explained by aging over 24 months. PMID- 12777367 TI - Childhood diarrhea and observed hygiene behavior in Salvador, Brazil. AB - Brief biweekly home visits, made as part of a cohort study of diarrhea in young children under age 5 years that was carried out in Salvador, Brazil, in 1998 1999, were used as a low-cost way to collect structured observation data on domestic hygiene behavior. Field-workers were trained to check a list of 23 forms of hygienic or unhygienic behavior by the child or the child's caretaker, if any behaviors were seen during the visit. Children were grouped according to whether mainly unhygienic behavior or mainly hygienic behavior had been recorded. This permitted study of the determinants of hygiene behavior and of its role in the transmission or prevention of diarrheal disease. Observations were recorded on roughly one visit in 20. Households with adequate excreta disposal were significantly more likely to be in the "mainly hygienic" group. The prevalence of diarrhea among children for whom mainly unhygienic behavior was recorded was 2.2 times that among children in the "mainly hygienic" group. The relative risk for prevalence was 2.2 (95% confidence interval: 1.7, 2.8). The relative risk fell to 1.9 (95% confidence interval: 1.5, 2.5) after data were controlled for confounding, but the difference was still highly significant. PMID- 12777369 TI - Re: "Blood transfusions as a risk factor for non-Hodgkins lymphoma in the San Francisco Bay area: a population based study". PMID- 12777368 TI - Longitudinal evaluation of an educational intervention for preventing tick bites in an area with endemic lyme disease in Baltimore County, Maryland. AB - The authors attempted to determine whether a targeted educational intervention in an area with endemic Lyme disease could increase knowledge, positive attitudes, and reported behaviors related to tick bite prevention and consequently decrease tick bites, as measured by a biomarker of tick bites. Between April and September of 1999, 317 subjects in Baltimore County, Maryland, were randomized to receive either tick-related or general health-related educational materials bimonthly through the mail. At each of three clinic visits, participants completed a self administered questionnaire and provided a serum sample. Anti-recombinant tick calreticulin antibody (ARTCA), measured in ng/ micro l, was used as a biomarker of tick bites. Linear and logistic regression analyses were used to determine 1) whether the educational intervention was associated with a change in knowledge, attitudes, and behaviors (KAB) and 2) whether change in KAB predicted change in ARTCA levels. Proportions of desired responses increased significantly among intervention subjects versus the comparison group on KAB measures related to examining the body for ticks and insect repellent use. Levels of ARTCA were low among all study subjects. Only six of 37 models exhibited a significant relation between change in a KAB variable and change in ARTCA levels over time. The behavioral intervention was associated with an increase in the KAB measures in the intervention group, but this change was not associated with change in ARTCA levels. PMID- 12777371 TI - Abeta42-lowering nonsteroidal anti-inflammatory drugs preserve intramembrane cleavage of the amyloid precursor protein (APP) and ErbB-4 receptor and signaling through the APP intracellular domain. AB - Epidemiological studies indicate that long term use of nonsteroidal anti inflammatory drugs (NSAIDs) confers protection from Alzheimer's disease, and some NSAIDs were shown to specifically decrease production of the amyloidogenic Abeta42 peptide, most likely by direct modulation of gamma-secretase activity. In contrast to gamma-secretase inhibitors, Abeta42-lowering NSAIDs do not impair S3 cleavage in the NOTCH receptor and release of the NOTCH intracellular domain, a finding with conceptual implications for the development of safer drugs targeting Abeta production through gamma-secretase modulation. Intramembrane cleavage and release of an intracellular signaling domain has recently been demonstrated in a number of additional gamma-secretase substrates. We now show in cell-based assays that intramembrane cleavage of APP and ErbB-4 receptor is not impaired by the Abeta42-lowering NSAIDs, sulindac sulfide and ibuprofen. Generation of the APP intracellular domain (AICD) was further not inhibited in a cell-free assay at concentrations far exceeding those effective in reducing Abeta42 production. Closer inspection of AICD signaling showed that stabilization of the AICD peptide by FE65 and AICD-mediated transcription were also retained at Abeta42-lowering concentrations. These results demonstrate that S3-like/intramembrane cleavage is preserved by Abeta42-lowering NSAIDs in at least three substrates of gamma secretase APP, ErbB-4, and NOTCH and underline the striking specificity by which these drugs target Abeta42 production. PMID- 12777372 TI - A novel hypoxia-inducible factor-independent hypoxic response regulating mammalian target of rapamycin and its targets. AB - Hypoxia triggers a reversible inhibition of protein synthesis thought to be important for energy conservation in O2-deficient environments. The mammalian target of rapamycin (mTOR) pathway integrates multiple environmental cues to regulate translation in response to nutrient availability and stress, suggesting it as a candidate for O2 regulation. We show here that hypoxia rapidly and reversibly triggers hypophosphorylation of mTOR and its effectors 4E-BP1, p70S6K, rpS6, and eukaryotic initiation factor 4G. Hypoxic regulation of these translational control proteins is dominant to activation via multiple distinct signaling pathways such as insulin, amino acids, phorbol esters, and serum and is independent of Akt/protein kinase B and AMP-activated protein kinase phosphorylation, ATP levels, ATP:ADP ratios, and hypoxia-inducible factor-1 (HIF 1). Finally, hypoxia appears to repress phosphorylation of translational control proteins in a manner analogous to rapamycin and independent of phosphatase 2A (PP2A) activity. These data demonstrate a new mode of regulation of the mTOR pathway and position this pathway as a powerful point of control by O2 of cellular metabolism and energetics. PMID- 12777373 TI - Receptors involved in the oxidized 1-palmitoyl-2-arachidonoyl-sn-glycero-3 phosphorylcholine-mediated synthesis of interleukin-8. A role for Toll-like receptor 4 and a glycosylphosphatidylinositol-anchored protein. AB - We demonstrated previously that oxidized 1-palmitoyl-2-arachidonoyl-sn-glycero-3 phosphorylcholine (ox-PAPC) and, specifically, the component lipid 1-palmitoyl-2 (5,6-epoxyisoprostane E2)-sn-glycero-3-phosphorylcholine increase interleukin-8 (IL-8) synthesis in aortic endothelial cells. The goal of the current studies was to characterize the receptor complex mediating the increased transcription of IL 8. We demonstrate that scavenger receptor class A, types I and II, lectin-like ox LDL receptor-1, macrophage receptor with collagenous structure, and CD36 are not responsible for the increase in IL-8. Using dominant-negative constructs and antisense oligonucleotides, we demonstrate a role for Toll-like receptor 4 (TLR4) as the ox-PAPC receptor mediating IL-8 transcription. We demonstrate that a glycosylphosphatidylinositol (GPI)-anchored protein is also necessary because phosphatidylinositol-specific phospholipase C pretreatment inhibited the effect of ox-PAPC. CD14, a GPI-anchored protein that associates with TLR4 in mediating lipopolysaccharide action, did not appear to mediate ox-PAPC action because ox PAPC-induced IL-8 transcription was not blocked by anti-CD14 neutralizing antibodies nor was it augmented by the addition of soluble CD14 or overexpression of membrane CD14. Instead, anti-TLR4 antibodies immunoprecipitated a 37-kDa protein that also bound ox-PAPC. A protein of this same size was found in aerolysin overlays used to detect GPI-anchored proteins. Therefore, these studies suggest that ox-PAPC may initially bind to a 37-kDa GPI-anchored protein, which interacts with TLR4 to induce IL-8 transcription. PMID- 12777374 TI - DNA binding activity of cytoplasmic phosphorylated Stat6 is masked by an interaction with a detergent-sensitive factor. AB - Signal transducer and activator of transcription (Stat) 6 is vital to interleukin (IL)-4 and IL-13 responses and the generation of Th2 immunity. We investigated the cellular location of phosphorylated Stat6 and Stat6 DNA binding activity in A201.1 murine B cells and primary splenocytes. Phosphorylated Stat6 was present in cytoplasmic and nuclear extracts from IL-4-treated cells. Confocal microscopy confirmed the presence of phosphorylated Stat6 in the cytoplasm of IL-4-treated cells. In contrast, Stat6 DNA binding activity was present in nuclear extracts, but not in cytoplasmic extracts. Thus, cytoplasmic extracts from IL-4-stimulated cells were devoid of Stat6 DNA binding activity despite the presence of phosphorylated Stat6. Addition of cytoplasmic extracts to nuclear extracts did not inhibit Stat6 DNA binding present in the nuclear extracts. Detergent treatment restored Stat6 DNA binding activity in cytoplasmic extracts of IL-4 stimulated cells. Thus, DNA binding activity of cytoplasmic phosphorylated Stat6 is masked by a factor dissociable by detergent treatment. PMID- 12777375 TI - Glucose-6-phosphate dehydrogenase modulates vascular endothelial growth factor mediated angiogenesis. AB - Glucose-6-phosphate dehydrogenase (G6PD), the first enzyme of the pentose phosphate pathway, is the principal intracellular source of NADPH. NADPH is utilized as a cofactor by vascular endothelial cell nitric-oxide synthase (eNOS) to generate nitric oxide (NO*). To determine whether G6PD modulates NO*-mediated angiogenesis, we decreased G6PD expression in bovine aortic endothelial cells using an antisense oligodeoxynucleotide to G6PD or increased G6PD expression by adenoviral gene transfer, and we examined vascular endothelial growth factor (VEGF)-stimulated endothelial cell proliferation, migration, and capillary-like tube formation. Deficient G6PD activity was associated with a significant decrease in endothelial cell proliferation, migration, and tube formation, whereas increased G6PD activity promoted these processes. VEGF-stimulated eNOS activity and NO* production were decreased significantly in endothelial cells with deficient G6PD activity and enhanced in G6PD-overexpressing cells. In addition, G6PD-deficient cells demonstrated decreased tyrosine phosphorylation of the VEGF receptor Flk-1/KDR, Akt, and eNOS compared with cells with normal G6PD activity, whereas overexpression of G6PD enhanced phosphorylation of Flk-1/KDR, Akt, and eNOS. In the Pretsch mouse, a murine model of G6PD deficiency, vessel outgrowth from thoracic aorta segments was impaired compared with C3H wild-type mice. In an in vivo Matrigel angiogenesis assay, cell migration into the plugs was inhibited significantly in G6PD-deficient mice compared with wild-type mice, and gene transfer of G6PD restored the wild-type phenotype in G6PD-deficient mice. These findings demonstrate that G6PD modulates angiogenesis and may represent a novel angiogenic regulator. PMID- 12777376 TI - Mechanistic studies on the intramolecular one-electron transfer between the two flavins in the human neuronal nitric-oxide synthase and inducible nitric-oxide synthase flavin domains. AB - Neuronal nitric-oxide synthase (nNOS) differs from inducible NOS (iNOS) in both its dependence on the intracellular Ca2+ concentration and the production rate of NO. To investigate what difference(s) exist between the two NOS flavin domains at the electron transfer level, we isolated the recombinant human NOS flavin domains, which were co-expressed with human calmodulin (CaM). The flavin semiquinones, FADH* and FMNH*, in both NOSs participate in the regulation of one electron transfer within the flavin domain. Each semiquinone can be identified by a characteristic absorption peak at 520 nm (Guan, Z.-W., and Iyanagi, T. (2003) Arch. Biochem. Biophys. 412, 65-76). NADPH reduction of the FAD and FMN redox centers by the CaM-bound flavin domains was studied by stopped-flow and rapid scan spectrometry. Reduction of the air-stable semiquinone (FAD-FMNH*) of both domains with NADPH showed that the extent of conversion of FADH2/FMNH* to FADH*/FMNH2 in the iNOS flavin domain was greater than that of the nNOS flavin domain. The reduction of both oxidized domains (FAD-FMN) with NADPH resulted in the initial formation of a small amount of disemiquinone, which then decayed. The rate of intramolecular electron transfer between the two flavins in the iNOS flavin domain was faster than that of the nNOS flavin domain. In addition, the formation of a mixture of the two- and four-electron-reduced states in the presence of excess NADPH was different for the two NOS flavin domains. The data indicate a more favorable formation of the active intermediate FMNH2 in the iNOS flavin domain. PMID- 12777377 TI - Insulin-like growth factor-binding protein-5 inhibits the growth of human breast cancer cells in vitro and in vivo. AB - The role of insulin-like growth factor-binding protein (IGFBP)-5 in human breast cancer cell growth is unclear. We determined the effects of IGFBP-5 expression on the growth of human breast cancer cell lines in vivo and in vitro. Expression of IGFBP-5, both by stable transfection and adenoviral-mediated infection, was inhibitory to the growth of MDA-MB-231 and Hs578T human breast cancer cells over a 13-day period. IGFBP-5 expression resulted in a G2/M cell cycle arrest and the induction of apoptosis in both cell lines, an effect that was abrogated in the presence of the broad-spectrum caspase inhibitor, z-VAD-fmk. IGFBP-5-induced apoptosis was associated with a transcriptional increase in expression of the proapoptotic regulator bax and decrease in the anti-apoptotic bcl-2 compared with vector controls. Secreted IGFBP-5 when added exogenously to breast cancer cells was not internalized and had no effect on cell growth or apoptosis, suggesting that IGFBP-5 may elicit its inhibitory effects via a novel, intracrine mechanism. In athymic nude mice, stable expression of IGFBP-5 significantly inhibited both the formation and growth of tumors derived from MDA-MB-231 cells. IGFBP-5 expressing tumors also had a significantly elevated level of bax mRNA and decreased levels of bcl-2 mRNA compared with vector tumors. These data suggest that IGFBP-5 is a potent growth inhibitor and proapoptotic agent in human breast cancer cells via modulation of cell cycle regulation and apoptotic mediators. PMID- 12777378 TI - Dual specificity mitogen-activated protein (MAP) kinase phosphatase-4 plays a potential role in insulin resistance. AB - Insulin is the key hormone that controls glucose homeostasis. Dysregulation of insulin function causes diabetes mellitus. Among the two major forms of diabetes, type 2 diabetes accounts for over 90% of the affected population. The incidence of type 2 diabetes is highly related to obesity. To find novel proteins potentially involved in obesity-related insulin resistance and type 2 diabetes, a functional expression screen was performed to search for genes that negatively regulate insulin signaling. Specifically, a reporter system comprised of the PEPCK promoter upstream of alkaline phosphatase was used in a hepatocyte cell based assay to screen an expression cDNA library for genes that reverse insulin induced repression of PEPCK transcription. The cDNA library used in this study was derived from the white adipose tissue of ob/ob mice, which are highly insulin resistant. The mitogen-activated dual specificity protein kinase phosphatase 4 (MKP-4) was identified as a candidate gene in this screen. Here we show that MKP 4 is expressed in insulin-responsive tissues and that the expression levels are up-regulated in obese insulin-resistant rodent models. Heterologous expression of MKP-4 in preadipocytes significantly blocked insulin-induced adipogenesis, and overexpression of MKP-4 in adipocytes inhibited insulin-stimulated glucose uptake. Our data suggest that MKP-4 negatively regulates insulin signaling and, consequently, may contribute to the pathogenesis of insulin resistance. PMID- 12777379 TI - Peptides induce ATP hydrolysis at both subunits of the transporter associated with antigen processing. AB - The transporter associated with antigen processing (TAP) plays a key role in the adaptive immune response by pumping antigenic peptides into the endoplasmic reticulum for subsequent loading of major histocompatibility complex class I molecules. TAP is a heterodimer consisting of TAP1 and TAP2. Each subunit is composed of a transmembrane domain and a nucleotide-binding domain, which energizes the peptide transport. To analyze ATP hydrolysis of each subunit we developed a method of trapping 8-azido-nucleotides to TAP in the presence of phosphate transition state analogs followed by photocross-linking, immunoprecipitation, and high resolution SDS-PAGE. Strikingly, trapping of both TAP subunits by beryllium fluoride is peptide-specific. The peptide concentration required for half-maximal trapping is identical for TAP1 and TAP2 and directly correlates with the peptide binding affinity. Only a background level of trapping was observed for low affinity peptides or in the presence of the herpes simplex viral protein ICP47, which specifically blocks peptide binding to TAP. Importantly, the peptide-induced trapped state is reached after ATP hydrolysis and not in a backward reaction of ADP binding and trapping. In the trapped state, TAP can neither bind nor exchange nucleotides, whereas peptide binding is not affected. In summary, these data support the model that peptide binding induces a conformation that triggers ATP hydrolysis in both subunits of the TAP complex within the catalytic cycle. PMID- 12777380 TI - Assembly of the yeast prion Ure2p into protein fibrils. Thermodynamic and kinetic characterization. AB - The [URE3] phenotype in Saccharomyces cerevisiae propagates by a prion mechanism, involving the aggregation of the normally soluble and highly helical protein Ure2. Previous data have shown that the protein spontaneously forms in vitro long, straight, insoluble fibrils at neutral pH that are similar to amyloids in that they bind Congo red and show green-yellow birefringence and have an increased resistance to proteolysis. These fibrils are not amyloids as they are devoid of a cross-beta core. Here we further document the mechanism of assembly of Ure2p into fibrils. The critical concentration for Ure2p assembly is measured, and the minimal size of the nuclei that are the precursors of Ure2p fibrils is determined. Our data indicate that the assembly process is irreversible. As a consequence, the critical concentration is very low. By analyzing the elongation rates of preformed fibrils and combining the results with single-fiber imaging experiments of a variant Ure2p labeled by fluorescent dyes, we reveal the polarity of the fibrils and differences in the elongation rates at their ends. These results bring novel insight in the process of Ure2p assembly into fibrils and the mechanism of propagation of yeast prions. PMID- 12777381 TI - Assembly and topography of the prepore complex in cholesterol-dependent cytolysins. AB - Cholesterol-dependent cytolysins are a family of poreforming proteins that have been shown to be virulence factors for a large number of pathogenic bacteria. The mechanism of pore formation for these toxins involves a complex series of events that are known to include binding, oligomerization, and insertion of a transmembrane beta-barrel. Several features of this mechanism remain poorly understood and controversial. Whereas a prepore mechanism has been proposed for perfringolysin O, a very different mechanism has been proposed for the homologous member of the family, streptolysin O. To distinguish between the two models, a novel approach that directly measures the dimension of transmembranes pores was used. Pore formation itself was examined for both cytolysins by encapsulating fluorescein-labeled peptides and proteins of different sizes into liposomes. When these liposomes were re-suspended in a solution containing anti-fluorescein antibodies, toxin-mediated pore formation was monitored directly by the quenching of fluorescein emission as the encapsulated molecules were released, and the dyes were bound by the antibodies. The analysis of pore formation determined using this approach reveals that only large pores are produced by perfringolysin O and streptolysin O during insertion (and not small pores that grow in size). These results are consistent only with the formation of a prepore complex intermediate prior to insertion of the transmembrane beta-barrel into the bilayer. Fluorescence quenching experiments also revealed that PFO in the prepore complex contacts the membrane via domain 4, and that the individual transmembrane beta hairpins in domain 3 are not exposed to the nonpolar core of the bilayer at this intermediate stage. PMID- 12777382 TI - CloR, a bifunctional non-heme iron oxygenase involved in clorobiocin biosynthesis. AB - The aminocoumarin antibiotics novobiocin and clorobiocin contain a 3 dimethylallyl-4-hydroxybenzoate (3DMA-4HB) moiety. The biosynthesis of this moiety has now been identified by biochemical and molecular biological studies. CloQ from the clorobiocin biosynthetic gene cluster in Streptomyces roseochromogenes DS 12976 has recently been identified as a 4 hydroxyphenylpyruvate-3-dimethylallyltransferase. In the present study, the enzyme CloR was overexpressed in Escherichia coli, purified, and identified as a bifunctional non-heme iron oxygenase, which converts 3-dimethylallyl-4 hydroxyphenylpyruvate (3DMA-4HPP) via 3-dimethylallyl-4-hydroxymandelic acid (3DMA-4HMA) to 3DMA-4HB by two consecutive oxidative decarboxylation steps. In 18O2 labeling experiments we showed that two oxygen atoms are incorporated into the intermediate 3DMA-4HMA in the first reaction step, but only one further oxygen is incorporated into the final product 3DMA-4HB during the second reaction step. CloR does not show sequence similarity to known oxygenases. It apparently presents a novel member of the diverse family of the non-heme iron (II) and alpha ketoacid-dependent oxygenases, with 3DMA-4HPP functioning both as an alpha-keto acid and as a hydroxylation substrate. The reaction catalyzed by CloR represents a new pathway for the formation of benzoic acids in nature. PMID- 12777383 TI - Repression of the human adenine nucleotide translocase-2 gene in growth-arrested human diploid cells: the role of nuclear factor-1. AB - Adenine nucleotide translocase-2 (ANT2) catalyzes the exchange of ATP for ADP across the mitochondrial membrane, thus playing an important role in maintaining the cytosolic phosphorylation potential required for cell growth. Expression of ANT2 is activated by growth stimulation of quiescent cells and is down-regulated when cells become growth-arrested. In this study, we address the mechanism of growth arrest repression. Using a combination of transfection, in vivo dimethyl sulfate mapping, and in vitro DNase I mapping experiments, we identified two protein-binding elements (Go-1 and Go-2) that are responsible for growth arrest of ANT2 expression in human diploid fibroblasts. Proteins that bound the Go elements were purified and identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry as members of the NF1 family of transcription factors. Chromatin immunoprecipitation analysis showed that NF1 was bound to both Go-1 and Go-2 in quiescent human diploid cells in vivo, but not in the same cells stimulated to growth by serum. NF1 binding correlated with the disappearance of ANT2 transcripts in quiescent cells. Furthermore, overexpression of NF1-A, -C, and -X in NIH3T3 cells repressed expression of an ANT2-driven reporter gene construct. Two additional putative repressor elements in the ANT2 promoter, an Sp1 element juxtaposed to the transcription start site and a silencer centered at nucleotide -332, did not appear to contribute to growth arrest repression. Thus, enhanced binding of NF1 is a key step in the growth arrest repression of ANT2 transcription. To our knowledge, this is the first report showing a role for NF1 in growth arrest. PMID- 12777384 TI - ARP-1/COUP-TF II determines hepatoma phenotype by acting as both a transcriptional repressor of microsomal triglyceride transfer protein and an inducer of CYP7A1. AB - L35 and FAO cells were derived as single cell isolates from H35 cells. Whereas L35 cells do not express microsomal triglyceride transfer protein (MTP), which regulates lipoprotein secretion, they express CYP7A1, which regulates bile acid synthesis from cholesterol. FAO cells display the opposite phenotype (i.e. expression of MTP but not CYP7A1). We examined the molecular basis of the transcriptional inactivation of the MTP gene in L35 cells. Nested deletion and mutagenesis studies show that a conserved DR1 element within the 135-bp proximal MTP promoter is responsible for differential expression by L35 and FAO cells. Yeast one-hybrid screening identified apolipoprotein A1 regulatory protein 1/chicken ovalbumin upstream promoter transcription factor II (ARP-1/COUP-TFII) and retinoid X receptor (RXRalpha) as the protein factors that can bind to the conserved DR1 element. Nuclear extracts from L35 cells contained 2-fold more ARP 1/COUP-TFII and 50% less RXRalpha than those from FAO cells. Immunologic studies show that in L35 cells, ARP-1/COUP-TFII is bound to the DR1 element, whereas in FAO cells, a complex containing RXRalpha is bound to the DR1 element. Co transfection studies show that ARP-1/COUP-TFII repressed MTP promoter activity by approximately 70% in FAO hepatoma cells, whereas RXRalpha and its ligand 9-cis retinoic acid increased MTP promoter activity by 6-fold in L35 cells. The combined data suggest that in the context of the MTP promoter, ARP-1/COUP-TFII (repressor) and a complex containing RXRalpha (inducer) compete for the DR1 element. Analysis of the CYP7A1 promoter revealed that it is approximately 5-fold more active in L35 cells than in FAO cells. Co-transfection of an ARP-1/COUP-TFII expression vector showed that it enhances CYP7A1 promoter activity by 6-fold in FAO cells. These combined findings indicate that ARP-1/COUP-TFII acts as both a transcriptional repressor (of MTP) and as a transcription activator (of CYP7A1). This dual function of ARP-1/COUP-TFII may play an important role in determining the metabolic phenotype of individual liver cells. PMID- 12777385 TI - Proteomic analysis of human Nop56p-associated pre-ribosomal ribonucleoprotein complexes. Possible link between Nop56p and the nucleolar protein treacle responsible for Treacher Collins syndrome. AB - Nop56p is a component of the box C/D small nucleolar ribonucleoprotein complexes that direct 2'-O-methylation of pre-rRNA during its maturation. Genetic analyses in yeast have shown that Nop56p plays important roles in the early steps of pre rRNA processing. However, its precise function remains elusive, especially in higher eukaryotes. Here we describe the proteomic characterization of human Nop56p (hNop56p)-associated pre-ribosomal ribonucleoprotein complexes. Mass spectrometric analysis of purified pre-ribosomal ribonucleoprotein complexes identified 61 ribosomal proteins, 16 trans-acting factors probably involved in ribosome biogenesis, and 29 proteins whose function in ribosome biogenesis is unknown. Identification of pre-rRNA species within hNop56p-associated pre ribosomal ribonucleoprotein complexes, coupled with the known functions of yeast orthologs of the probable trans-acting factors identified in human, demonstrated that hNop56p functions in the early to middle stages of 60 S subunit synthesis in human cells. Interestingly, the nucleolar phosphoprotein treacle, which is responsible for the craniofacial disorder associated with Treacher Collins syndrome, was found to be a constituent of hNop56p-associated pre-rRNP complexes. The association of hNop56p and treacle within the complexes was independent of rRNA integrity, indicating a direct interaction. In addition, the protein compositions of the treacle-associated and hNop56p-associated pre-ribosomal ribonucleoprotein complexes were very similar, suggesting functional similarities between these two complexes with respect to ribosome biogenesis in human cells. PMID- 12777387 TI - Functional diversity of the Drosophila PGRP-LC gene cluster in the response to lipopolysaccharide and peptidoglycan. AB - The peptidoglycan recognition protein PGRP-LC is a major activator of the imd/Relish pathway in the Drosophila immune response. Three transcripts are generated by alternative splicing of the complex PGRP-LC gene. The encoded transmembrane proteins share an identical intracellular part, but each has a separate extracellular PGRP-domain: x, y, or a. Here we show that two of these isoforms play unique roles in the response to different microorganisms. Using RNA interference in Drosophila mbn-2 cells, we found that PGRP-LCx is the only isoform required to mediate signals from Gram-positive bacteria and purified bacterial peptidoglycan. By contrast, the recognition of Gram-negative bacteria and bacterial lipopolysaccharide requires both PGRP-LCa and LCx. The third isoform, LCy, is expressed at lower levels and may be partially redundant. Two additional PGRP domains in the gene cluster, z and w, are both included in a single transcript of a separate gene, PGRP-LF. Suppression of this transcript does not block the response to any of the microorganisms tested. PMID- 12777386 TI - High glucose augments the angiotensin II-induced activation of JAK2 in vascular smooth muscle cells via the polyol pathway. AB - Angiotensin II (Ang II), protein kinase C (PKC), reactive oxygen species (ROS) generated by NADPH oxidase, the activation of Janus kinase 2 (JAK2), and the polyol pathway play important parts in the hyperproliferation of vascular smooth muscle cells (VSMC), a characteristic feature of diabetic macroangiopathy. The precise mechanism, however, remains unclear. This study investigated the relation between the polyol pathway, PKC-beta, ROS, JAK2, and Ang II in the development of diabetic macroangiopathy. VSMC cultured in high glucose (HG; 25 mm) showed significant increases in the tyrosine phosphorylation of JAK2, production of ROS, and proliferation activities when compared with VSMC cultured in normal glucose (5.5 mm (NG)). Both the aldose reductase specific inhibitor (zopolrestat) or transfection with aldose reductase antisense oligonucleotide blocked the phosphorylation of JAK2, the production of ROS, and proliferation of VSMC induced by HG, but it had no effect on the Ang II-induced activation of these parameters in both NG and HG. However, transfection with PKC-beta antisense oligonucleotide, preincubation with a PKC-beta-specific inhibitor (LY379196) or apocynin (NADPH oxidase-specific inhibitor), or electroporation of NADPH oxidase antibodies blocked the Ang II-induced JAK2 phosphorylation, production of ROS, and proliferation of VSMC in both NG and HG. These observations suggest that the polyol pathway hyperactivity induced by HG contributes to the development of diabetic macroangiopathy through a PKC-beta-ROS activation of JAK2. PMID- 12777388 TI - Structural comparison of human monoamine oxidases A and B: mass spectrometry monitoring of cysteine reactivities. AB - Monoamine oxidases (MAO) A and B are approximately 60-kDa outer mitochondrial membrane flavoenzymes catalyzing the degradation of neurotransmitters and xenobiotic arylalkyl amines. Despite 70% identity of their amino acid sequences, both enzymes exhibit strikingly different properties when exposed to thiol modifying reagents. Human MAO A and MAO B each contain 9 cysteine residues (7 in conserved sequence locations). MAO A is inactivated by N-ethylmaleimide (NEM) much faster (tau(1/2) = approximately 3 min) than MAO B (tau(1/2) = approximately 8 h). These differences in thiol reactivities are also demonstrated by monitoring the NEM modification stoichiometries by electrospray mass spectrometry. Inactivation of either enzyme with acetylenic inhibitors results in alterations of their thiol reactivities. Cys5 and Cys266 were identified as the only residues modified by biotin-derivatized NEM in clorgyline-inactivated MAO A and pargyline inactivated MAO B, respectively. The x-ray structure of MAO B (Binda, C., Newton Vinson, P., Hubalek, F., Edmondson, D. E., and Mattevi, A. (2002) Nat. Struct. Biol. 9, 22-26) shows that Cys5 is located on the surface of the molecule opposite to the membrane-binding region. Cys266 in MAO A is predicted to be located in the same region of the molecule. These thiol residues are also modified by biotin-derivatized NEM in the mitochondrial membrane-bound MAO A and MAO B. This study shows that the MAO A structure is "more flexible" than that of MAO B and that clorgyline and pargyline inactivation of MAO A and B, respectively, increases the structural stability of both enzymes. No evidence is found for the presence of disulfide bonds in either enzyme, contrary to a previous suggestion. PMID- 12777389 TI - An electrical potential in the access channel of catalases enhances catalysis. AB - Substrate H2O2 must gain access to the deeply buried active site of catalases through channels of 30-50 A in length. The most prominent or main channel approaches the active site perpendicular to the plane of the heme and contains a number of residues that are conserved in all catalases. Changes in Val169, 8 A from the heme in catalase HPII from Escherichia coli, introducing smaller, larger or polar side chains reduces the catalase activity. Changes in Asp181, 12 A from the heme, reduces activity by up to 90% if the negatively charged side chain is removed when Ala, Gln, Ser, Asn, or Ile are the substituted residues. Only the D181E variant retains wild type activity. Determination of the crystal structures of the Glu181, Ala181, Ser181, and Gln181 variants of HPII reveals lower water occupancy in the main channel of the less active variants, particularly at the position forming the sixth ligand to the heme iron and in the hydrophobic, constricted region adjacent to Val169. It is proposed that an electrical potential exists between the negatively charged aspartate (or glutamate) side chain at position 181 and the positively charged heme iron 12 A distant. The potential field acts upon the electrical dipoles of water generating a common orientation that favors hydrogen bond formation and promotes interaction with the heme iron. Substrate hydrogen peroxide would be affected similarly and would enter the active site oriented optimally for interaction with active site residues. PMID- 12777390 TI - Localization of the deoxyribose phosphate lyase active site in human DNA polymerase iota by controlled proteolysis. AB - Human DNA polymerase iota (pol iota) is a member of the Y-family of low fidelity lesion bypass DNA polymerases. In addition to a probable role in DNA lesion bypass, this enzyme has recently been shown to be required for somatic hypermutation in human B-cells. We found earlier that human pol iota has deoxyribose phosphate (dRP) lyase activity and unusual specificity for activity during DNA synthesis, suggesting involvement in specialized forms of base excision repair (BER). Here, mapping of the domain structure of human pol iota by controlled proteolysis revealed that the enzyme has a 48-kDa NH2-terminal domain and a protease resistant 40-kDa "core domain" spanning residues Met79 to approximately Met445. A covalently cross-linked pol iota-DNA complex, representing a trapped intermediate in the dRP lyase reaction, was subjected to controlled proteolysis. Cross-linking was mapped to the 40-kDa core domain, indicating that the dRP lyase active site is in this region. To further evaluate the BER capacity of the enzyme, the dRP lyase and DNA polymerase activities were characterized on DNA substrates representing BER intermediates, and we found that pol iota was able to complement the in vitro single-nucleotide BER deficiency of a DNA polymerase beta null cell extract. PMID- 12777391 TI - Peroxisome proliferator-activated receptor-gamma represses GLUT4 promoter activity in primary adipocytes, and rosiglitazone alleviates this effect. AB - The synthetic thiazolidinedione ligands of peroxisome proliferator-activated receptor-gamma (PPARgamma) improve insulin sensitivity in type II diabetes and induce GLUT4 mRNA expression in fat and muscle. However, the molecular mechanisms involved are still unclear. We studied the regulatory effects of PPARgamma and its ligands on GLUT4 gene expression in primary rat adipocytes and CHO-K1 cells cotransfected with PPARgamma and the GLUT4 promoter reporter. PPARgamma1 and PPARgamma2 repressed the activity of the GLUT4 promoter in a dose-dependent manner. Whereas this repression was augmented by the natural ligand 15Delta prostaglandin J2, it was completely alleviated by rosiglitazone (Rg). Ligand binding-defective mutants PPARgamma1-L468A/E471A and PPARgamma2-L496A/E499A retained the repression effect, which was unaffected by Rg, whereas the PPARgamma2-S112A mutant exhibited a 50% reduced capacity to repress GLUT4 promoter activity. The -66/+163 bp GLUT4 promoter region was sufficient to mediate PPARgamma inhibitory effects. The PPARgamma/retinoid X receptor-alpha heterodimer directly bound to this region, whereas binding was abolished in the presence of Rg. Thus, we show that PPARgamma represses transcriptional activity of the GLUT4 promoter via direct and specific binding of PPARgamma/retinoid X receptor-alpha to the GLUT4 promoter. This effect requires an intact Ser112 phosphorylation site on PPARgamma and is completely alleviated by Rg, acting via its ligand-binding domain. These data suggest a novel mechanism by which Rg exerts its antidiabetic effects via detaching PPARgamma from the GLUT4 gene promoter, thus leading to increased GLUT4 expression and enhanced insulin sensitivity. PMID- 12777392 TI - Initiation and transduction of stretch-induced RhoA and Rac1 activation through caveolae: cytoskeletal regulation of ERK translocation. AB - The Rho family small GTPases play a crucial role in mediating cellular responses to stretch. However, it remains unclear how force is transduced to Rho signaling pathways. We investigated the effect of stretch on the activation and caveolar localization of RhoA and Rac1 in neonatal rat cardiomyocytes. In unstretched cardiomyocytes, RhoA and Rac1 were detected in both caveolar and non-caveolar fractions as assessed using detergent-free floatation analysis. Stretching myocytes for 4 min activated RhoA and Rac1. By 15 min of stretch, RhoA and Rac1 had dissociated from caveolae, and there was decreased coprecipitation of RhoA and Rac1 with caveolin-3. To determine whether compartmentation of RhoA and Rac1 within caveolae was necessary for stretch signaling, we disrupted caveolae with methyl beta-cyclodextrin (MbetaCD). Treatment with 5 mm MbetaCD for 1 h dissociated both RhoA and Rac1 from caveolae. Under this condition, stretch failed to activate RhoA or Rac1. Stretch-induced actin cytoskeletal organization was concomitantly impaired. Interestingly the ability of stretch to activate extracellular signal-regulated kinase (ERK) was unaffected by MbetaCD treatment, but ERK translocation to the nucleus was impaired. Stretch-induced hypertrophy was also inhibited. Actin cytoskeletal disruption with cytochalasin-D also prevented stretch from increasing nuclear ERK, whereas actin polymerization with jasplakinolide restored nuclear translocation of activated ERK in the presence of MbetaCD. We suggest that activation of RhoA or Rac1, localized in a caveolar compartment, is essential for sensing externally applied force and transducing this signal to the actin cytoskeleton and ERK translocation. PMID- 12777393 TI - Dok-R binds c-Abl and regulates Abl kinase activity and mediates cytoskeletal reorganization. AB - Dok-R, also known as Dok-2/FRIP, belongs to the DOK family of signaling molecules that become tyrosine-phosphorylated by several different receptor and cytoplasmic tyrosine kinases. Tyrosine phosphorylation of DOK proteins establishes high affinity binding sites for other signaling molecules leading to activation of a signaling cascade. Here we show that Dok-R associates with c-Abl directly via a constitutive SH3-mediated interaction and that this binding requires a PMMP motif in the proline-rich tail of Dok-R. The Dok-R-Abl interaction is further enhanced by an active c-Abl kinase, which requires the presence of its SH2 domain. Interaction of Dok-R with c-Abl also results in an increase in c-Abl tyrosine phosphorylation and kinase activity. Furthermore, we demonstrate that this increase in kinase activity correlates with a concomitant increase in c-Abl mediated biological activity as measured by the formation of actin microspikes. Our data are the first to demonstrate that Dok-R and c-Abl interact in both a constitutive and inducible fashion and that Dok-R influences the intracellular kinase and biological activity of c-Abl. PMID- 12777394 TI - Transgenic overexpression of the Ca2+-binding protein S100A1 in the heart leads to increased in vivo myocardial contractile performance. AB - S100A1, a Ca2+-sensing protein of the EF-hand family, is most highly expressed in myocardial tissue, and cardiac S100A1 overexpression in vitro has been shown to enhance myocyte contractile properties. To study the physiological consequences of S100A1 in vivo, transgenic mice were developed with cardiac-restricted overexpression of S100A1. Characterization of two independent transgenic mouse lines with approximately 4-fold overexpression of S100A1 in the myocardium revealed a marked augmentation of in vivo basal cardiac function that remained elevated after beta-adrenergic receptor stimulation. Contractile function and Ca2+ handling properties were increased in ventricular cardiomyocytes isolated from S100A1 transgenic mice. Enhanced cellular Ca2+ cycling by S100A1 was associated both with increased sarcoplasmic reticulum Ca2+ content and enhanced sarcoplasmic reticulum Ca2+-induced Ca2+ release, and S100A1 was shown to associate with the cardiac ryanodine receptor. No alterations in beta-adrenergic signal transduction or major cardiac Ca2+-cycling proteins occurred, and there were no signs of hypertrophy with chronic cardiac S100A1 overexpression. Our findings suggest that S100A1 plays an important in vivo role in the regulation of cardiac function perhaps through interacting with the ryanodine receptor. Because S100A1 protein expression is down-regulated in heart failure, increasing S100A1 expression in the heart may represent a novel means to augment contractility. PMID- 12777395 TI - The reported human NADsyn2 is ammonia-dependent NAD synthetase from a pseudomonad. AB - Nicotinamide-adenine dinucleotide (NAD+) synthetases catalyze the last step in NAD+ metabolism in the de novo, import, and salvage pathways that originate from tryptophan (or aspartic acid), nicotinic acid, and nicotinamide, respectively, and converge on nicotinic acid mononucleotide. NAD+ synthetase converts nicotinic acid adenine dinucleotide to NAD+ via an adenylylated intermediate. All of the known eukaryotic NAD+ synthetases are glutamine-dependent, hydrolyzing glutamine to glutamic acid to provide the attacking ammonia. In the prokaryotic world, some NAD+ synthetases are glutamine-dependent, whereas others can only use ammonia. Earlier, we noted a perfect correlation between presence of a domain related to nitrilase and glutamine dependence and then proved in the accompanying paper (Bieganowski, P., Pace, H. C., and Brenner, C. (2003) J. Biol. Chem. 278, 33049 33055) that the nitrilase-related domain is an essential, obligate intramolecular, thiol-dependent glutamine amidotransferase in the yeast NAD+ synthetase, Qns1. Independently, human NAD+ synthetase was cloned and shown to depend on Cys-175 for glutamine-dependent but not ammonia-dependent NAD+ synthetase activity. Additionally, it was claimed that a 275 amino acid open reading frame putatively amplified from human glioma cell line LN229 encodes a human ammonia-dependent NAD+ synthetase and this was speculated largely to mediate NAD+ synthesis in human muscle tissues. Here we establish that the so called NADsyn2 is simply ammonia-dependent NAD+ synthetase from Pseudomonas, which is encoded on an operon with nicotinic acid phosphoribosyltransferase and, in some Pseudomonads, with nicotinamidase. PMID- 12777396 TI - Somatic histones are components of the perinuclear theca in bovine spermatozoa. AB - The perinuclear theca is a non-ionic detergent-resistant, electron-dense layer surrounding the condensed nucleus of mammalian sperm. The known proteins originating from the perinuclear theca have implicated the structure in a variety of important cellular processes during spermiogenesis and fertilization. Nonetheless, the composition of the perinuclear theca remains largely unexplored. We have isolated a group of low molecular mass (14-19 kDa) perinuclear theca derived proteins from acrosome-depleted bovine sperm heads by salt (1 M KCl) extraction and have identified them as core somatic histones. N-terminal sequencing and immunoblotting with anti-histone antibodies confirmed the presence of both intact and proteolytically cleaved somatic histones H3, H2B, H2A, and H4. Identical proteins were isolated using 2% SDS or 1 N HCl extractions. Subsequent acid and SDS extractions of intact bovine sperm revealed the presence of all four intact histone subtypes, with minimal proteolysis. Two-dimensional acid/urea/Triton-SDS-PAGE, coupled with immunoblotting analysis, confirmed the somatic nature of these perinuclear theca-derived histones. Estimates of the abundance of perinuclear theca-derived histones showed that up to 0.2 pg per sperm of each histone subtype was present. Immunogold labeling at the ultrastructural level localized all four core somatic histones to the post acrosomal sheath region of bovine epididymal sperm, when probed with affinity purified anti-histone antibodies. Little immunoreactivity was detected in residual perinuclear theca structures following the extractions. Taken together, these findings indicate the unprecedented and stable localization of non-nuclear somatic histones in bovine sperm perinuclear theca. PMID- 12777397 TI - Overexpression of peroxisome proliferator-activated receptor gamma coactivator 1alpha down-regulates GLUT4 mRNA in skeletal muscles. AB - Exercise training increases mitochondria and GLUT4 in skeletal muscles. Recent studies indicate that an increased expression of the transcriptional coactivator peroxisome proliferator-activated receptor gamma coactivator 1alpha (PGC-1alpha) by exercise may promote mitochondrial biogenesis and fatty acid oxidation. To examine whether increased PGC-1alpha expression was also responsible for an increase of GLUT4 expression, transgenic mice that overexpress PGC-1alpha in skeletal muscles driven by a human alpha-skeletal actin promoter were made. PGC 1alpha was overexpresssed in skeletal muscles including type I and II fiber-rich muscles but not in the heart. With an increase of PGC-1alpha mRNA, type II fiber rich muscles were redder, and genes of mitochondrial oxidative metabolism were up regulated in skeletal muscles, whereas the expression of GLUT4 mRNA was unexpectedly down-regulated. In parallel with a decrease of GLUT4 mRNA, an impairment of glycemic control after intraperitoneal insulin administration was observed. Thus, an increase of PGC-1alpha plays a role in increasing mitochondrial biogenesis and fatty acid oxidation but not in increasing GLUT4 mRNA in skeletal muscles. PMID- 12777398 TI - Increased expression of cytochrome P450 2E1 induces heme oxygenase-1 through ERK MAPK pathway. AB - The inducible form of heme oxygenase (HO-1) is increased during oxidative injury, and this may be an important defense mechanism against such injury. Cytochrome P450 2E1 (CYP2E1) generates reactive oxygen species and promotes lipid peroxidation. In this study induction of HO-1 by CYP2E1 and the possible role of mitogen-activated protein kinase (MAPK) in this process were evaluated. HO-1 induction was observed in the livers of chronic alcohol-fed mice or pyrazole treated rats, conditions known to elevate CYP2E1 levels. Increased levels of HO-1 were observed in HepG2 cells overexpressing CYP2E1 (E47 cells) compared with control HepG2 cells or HepG2 cells expressing CYP3A4. Expression of CYP2E1 in HepG2 cells transcriptionally activated the HO-1 gene, increasing HO-1 mRNA and protein expression and activity of a HO-1 reporter construct. CYP2E1 inhibitors and catalase blocked the increased production of reactive oxygen species as well as HO-1 induction. Increasing oxidative stress by the addition of arachidonic acid or depletion of glutathione further increased HO-1 induction. The phosphorylated form of ERK MAPK but not that of p38 or JNK MAPK was increased in E47 cells compared with the control C34 HepG2 cells. PD98059, a specific inhibitor of ERK MAPK, blocked the activity of a HO-1 reporter in E47 cells but not in C34 cells. These results suggest that increased CYP2E1 activity leads to induction of the HO-1 gene, and the ERK MAPK pathway is important in mediating this process. This induction may serve as an adaptive mechanism to protect the E47 cells against the CYP2E1-dependent oxidative stress. PMID- 12777399 TI - Catalytic activity of ADAM8, ADAM15, and MDC-L (ADAM28) on synthetic peptide substrates and in ectodomain cleavage of CD23. AB - The ADAM family of disintegrin metalloproteases plays important roles in "ectodomain shedding," the process by which biologically active, soluble forms of cytokines, growth factors, and their receptors are released from membrane-bound precursors. Whereas ADAM8, ADAM15, and MDC-L (ADAM28) are expressed in specific cell types and tissues, their in vivo functions and substrates are not known. By screening a library of synthetic peptides as potential substrates, we show that soluble recombinant forms of these enzymes have similar proteolytic substrate specificity, clearly distinct from that of ADAM17 (TNFalpha-converting enzyme). A number of tumor necrosis factor (TNF) family proteins and CD23 were screened as potential substrates for ectodomain cleavage. We found that ADAM8, ADAM15, and MDC-L, but not ADAM17, catalyzed ectodomain shedding of CD23, the low affinity IgE receptor. ADAM8-dependent, soluble CD23 release required proteolytically active ADAM8, and a physical association of ADAM8 was observed with the membrane bound form of CD23. The ADAM8-dependent release of sCD23 and the endogenous release from B cell lines could be similarly inhibited by a hydroxamic acid, metalloprotease inhibitor compound. We conclude that ADAM8 could contribute to ectodomain shedding of CD23 and may thus be a potential target for therapeutic intervention in allergy and inflammation. PMID- 12777400 TI - Catalase activity is regulated by c-Abl and Arg in the oxidative stress response. AB - The Abl family of mammalian non-receptor tyrosine kinases includes c-Abl and Arg. Recent studies have demonstrated that c-Abl and Arg are activated in the response of cells to oxidative stress. This work demonstrates that catalase, a major effector of the cellular defense against H2O2, interacts with c-Abl and Arg. The results show that H2O2 induced binding of c-Abl and Arg to catalase. The SH3 domains of c-Abl and Arg bound directly to catalase at a P293FNP site. c-Abl and Arg phosphorylated catalase at Tyr231 and Tyr386 in vitro and in the response of cells to H2O2. The functional significance of the interaction is supported by the demonstration that cells deficient in both c-Abl and Arg exhibit substantial increases in H2O2 levels. In addition, c-abl-/- arg-/- cells exhibited a marked increase in H2O2-induced apoptosis compared with that found in the absence of either kinase. These findings indicate that c-Abl and Arg regulate catalase and that this signaling pathway is of importance to apoptosis in the oxidative stress response. PMID- 12777401 TI - A structural model for the open conformation of the mdr1 P-glycoprotein based on the MsbA crystal structure. AB - The validity of the structure of the Escherichia coli MsbA lipid transporter as a model from the mdr1 P-glycoprotein has been evaluated. Comparative sequence analyses, motif search and secondary structure prediction indicated that each of the two P-glycoprotein halves is structurally similar to the MsbA monomer and also suggested that the open dimer structure is valid for P-glycoprotein. Homology modeling was used to predict the structure of P-glycoprotein using MsbA as a template. The resulting modeled structure allowed a detailed study of the interactions between the intracellular domain and the nucleotide binding domain and suggested that these contacts are involved in mediating the coupling between nucleotide binding domain conformational changes and transmembrane helices reorientation during transport. In P-glycoprotein, the internal chamber open to the inner leaflet and the inner medium is significantly different in size and charge than in MsbA. These differences can be related to those of the transported substrates. Moreover an ensemble of 20 conserved aromatic residues appears to border the periphery of each side of the chamber in P-glycoprotein. These may be important for size selection and proper positioning of drugs for transport. The relevance of the modeled conformation to P-gp function is discussed. PMID- 12777402 TI - Quaternary structure of Azospirillum brasilense NADPH-dependent glutamate synthase in solution as revealed by synchrotron radiation x-ray scattering. AB - Azospirillum brasilense glutamate synthase (GltS) is the prototype of bacterial NADPH-dependent enzymes, a class of complex iron-sulfur flavoproteins essential in ammonia assimilation processes. The catalytically active GltS alpha beta holoenzyme and its isolated alpha and beta subunits (162 and 52 kDa, respectively) were analyzed using synchrotron radiation x-ray solution scattering. The GltS alpha subunit and alpha beta holoenzyme were found to be tetrameric in solution, whereas the beta subunit was a mixture of monomers and dimers. Ab initio low resolution shapes restored from the scattering data suggested that the arrangement of alpha subunits in the (alpha beta)4 holoenzyme is similar to that in the tetrameric alpha 4 complex and that beta subunits occupy the periphery of the holoenzyme. The structure of alpha 4 was further modeled using the available crystallographic coordinates of the monomeric alpha subunit assuming P222 symmetry. To model the entire alpha beta holoenzyme, a putative alpha beta protomer was constructed from the coordinates of the alpha subunit and those of the N-terminal region of porcine dihydropyrimidine dehydrogenase, which is similar to the beta subunit. Rigid body refinement yielded a model of GltS with an arrangement of alpha subunits similar to that in alpha 4, but displaying contacts also between beta subunits belonging to adjacent protomers. The holoenzyme model allows for independent catalytic activity of the alpha beta protomers, which is consistent with the available biochemical evidence. PMID- 12777403 TI - Dendritic cell immunoactivating receptor, a novel C-type lectin immunoreceptor, acts as an activating receptor through association with Fc receptor gamma chain. AB - An increasing number of C-type lectin receptors are being discovered on dendritic cells, but their signaling abilities and underlying mechanisms require further definition. Among these, dendritic cell immunoreceptor (DCIR) induces negative signals through an inhibitory immunoreceptor tyrosine-based inhibitory motif (ITIM) in its cytoplasmic tail. Here we identify a novel C-type lectin receptor, dendritic cell immunoactivating receptor (DCAR), whose extracellular lectin domain is highly homologous to that of DCIR. DCAR is expressed similarly in tissues to DCIR, but its short cytoplasmic portion lacks signaling motifs like ITIM. However, a positively charged arginine residue is present in the transmembrane region of the DCAR, which may explain its association with Fc receptor gamma chain and its stable expression on the cell surface. Furthermore, cross-linking of DCAR in the presence of gamma chain activates calcium mobilization and tyrosine phosphorylation of cellular proteins. These signals are mediated by the immunoreceptor tyrosine-based activating motif (ITAM) of the gamma chain. Thus, DCAR is closely related to DCIR, but it introduces activating signals into antigen-presenting cells through its physical and functional association with ITAM-bearing gamma chain. The identification of this activating immunoreceptor provides an example of signaling via a dendritic cell-expressed C type lectin receptor. PMID- 12777404 TI - Tracking of airway and tissue mechanics during TLC maneuvers in mice. AB - A tracking impedance estimation technique was developed to follow the changes in total respiratory impedance (Zrs) during slow total lung capacity maneuvers in six anesthetized and mechanically ventilated BALB/c mice. Zrs was measured with the wave-tube technique and pseudorandom forced oscillations at nine frequencies between 4 and 38 Hz during inflation from a transrespiratory pressure of 0-20 cmH2O and subsequent deflation, each lasting for approximately 20 s. Zrs was averaged for 0.125 s and fitted by a model featuring airway resistance (Raw) and inertance, and tissue damping and elastance (H). Lower airway conductance (Glaw) was linearly related to volume above functional residual capacity (V) between 0 and 75-95% maximum V, with a mean slope of dGlaw/dV = 13.6 +/- 4.6 cmH2O-1. s-1. The interdependence of Raw and H was characterized by two distinct and closely linear relationships for the low- and high-volume regions, separated at approximately 40% maximum V. Comparison of Raw with the highest-frequency resistance of the total respiratory system revealed a marked volume-dependent contribution of tissue resistance to total respiratory system resistance, resulting in the overestimation of Raw by 19 +/- 8 and 163 +/- 40% at functional residual capacity and total lung capacity, respectively, whereas the lowest frequency reactance was proportional to H; these findings indicate that single frequency resistance values may become inappropriate as surrogates of Raw when tissue impedance is changing. PMID- 12777405 TI - Airway contractility and smooth muscle Ca(2+) signaling in lung slices from different mouse strains. AB - To investigate the hypothesis that altered Ca2+ signaling in airway smooth muscle cells (SMCs) is responsible for airway hyperreactivity, we compared, with the use of confocal and phase-contrast microscopy, the airway contractility and Ca2+ changes in SMCs induced by acetylcholine (ACh) in lung slices from different mouse strains (A/J, Balb/C, and C3H/ HeJ). The airways from each mouse strain displayed a concentration-dependent contraction to ACh. The contractile response of the airways of the C3H/HeJ mice was found, in contrast to earlier studies, to be much greater and faster than that of A/J and Balb/C mice. This difference in airway reactivity can be, in part, attributable to halothane, a volatile anesthetic that was previously used during in vivo measurements of airway reactivity but found here to significantly alter the ACh contractile response of airways in lung slices. The ACh-induced Ca2+ response of the airway SMCs in all of the various mouse strains was also concentration dependent. The magnitude of the initial Ca2+ increase and the frequency of the subsequent Ca2+ oscillations induced by ACh increased with ACh concentration. However, no differences in the Ca2+ responses to ACh could be distinguished between the mouse strains. These results suggest that the mechanism responsible for airway hyperreactivity in different mouse strains resides with the Ca2+ sensitivity of the contractile apparatus of the SMCs rather than with the Ca2+ signaling itself. PMID- 12777406 TI - Effects of chronic AICAR treatment on fiber composition, enzyme activity, UCP3, and PGC-1 in rat muscles. AB - This study was designed to determine the histological and metabolic effects of the administration of 5'-AMP-activated protein kinase (AMPK) activator 5 aminoimidazole-4-carboxamide-1-beta-d-ribofuranoside (AICAR) for 14 successive days. AICAR treatment caused a significant decrease in the percentage of type IIB fibers and the concomitant increase in the percentage of type IIX fibers in extensor digitorum longus (EDL) muscle. The capillary density and the capillary to-fiber ratio were not altered by AICAR. AICAR treatment increased the glycolytic and oxidative enzyme activities but not the antioxidant enzyme activities. The AICAR treatment increased the uncoupling protein 3 (UCP3) level in EDL and the peroxisome proliferator-activated receptor-gamma coactivator 1alpha protein level in the soleus and EDL muscles, whereas the myogenin level was not altered by AICAR. These results seem to imply that the chronic activation of AMPK alters such muscle histochemical and metabolic characteristics. PMID- 12777407 TI - Cutaneous active vasodilation in humans during passive heating postexercise. AB - The hypothesis that exercise causes an increase in the postexercise esophageal temperature threshold for onset of cutaneous vasodilation through an alteration of active vasodilator activity was tested in nine subjects. Increases in forearm skin blood flow and arterial blood pressure were measured and used to calculate cutaneous vascular conductance at two superficial forearm sites: one with intact alpha-adrenergic vasoconstrictor activity (untreated) and one infused with bretylium tosylate (bretylium treated). Subjects remained seated resting for 15 min (no-exercise) or performed 15 min of treadmill running at either 55, 70, or 85% of peak oxygen consumption followed by 20 min of seated recovery. A liquid conditioned suit was used to increase mean skin temperature ( approximately 4.0 degrees C/h), while local forearm temperature was clamped at 34 degrees C, until cutaneous vasodilation. No differences in the postexercise threshold for cutaneous vasodilation between untreated and bretylium-treated sites were observed for either the no-exercise or exercise trials. Exercise resulted in an increase in the postexercise threshold for cutaneous vasodilation of 0.19 +/- 0.01, 0.39 +/- 0.02, and 0.53 +/- 0.02 degrees C above those of the no-exercise resting values for the untreated site (P < 0.05). Similarly, there was an increase of 0.20 +/- 0.01, 0.37 +/- 0.02, and 0.53 +/- 0.02 degrees C for the treated site for the 55, 70, and 85% exercise trials, respectively (P < 0.05). It is concluded that reflex activity associated with the postexercise increase in the onset threshold for cutaneous vasodilation is more likely mediated through an alteration of active vasodilator activity rather than through adrenergic vasoconstrictor activity. PMID- 12777408 TI - Mechanical ventilation-induced oxidative stress in the diaphragm. AB - Prolonged mechanical ventilation (MV) results in oxidative damage in the diaphragm; however, it is unclear whether this MV-induced oxidative injury occurs rapidly or develops slowly over time. Furthermore, it is unknown whether both soluble (cytosolic) and insoluble (myofibrillar) proteins are equally susceptible to oxidation during MV. These experiments tested two hypotheses: 1). MV-induced oxidative injury in the diaphragm occurs within the first 6 h after the initiation of MV; and 2). MV is associated with oxidative modification of both soluble and insoluble proteins. Adult Sprague-Dawley rats were randomly divided into one of seven experimental groups: 1) control (n = 8); 2) 3-h MV (n = 8); 3). 6-h MV (n = 6); 4). 18-h MV (n = 8); 5). 3-h anesthesia-spontaneous breathing (n = 8); 6). 6-h anesthesia-spontaneous breathing (n = 6); and 7). 18-h anesthesia spontaneous breathing (n = 8). Markers of oxidative injury in the diaphragm included the measurement of reactive (protein) carbonyl derivatives (RCD) and total lipid hydroperoxides. Three hours of MV did not result in oxidative injury in the diaphragm. In contrast, both 6 and 18 h of MV promoted oxidative injury in the diaphragm, as indicated by increases in both protein RCD and lipid hydroperoxides. Electrophoretic separation of soluble and insoluble proteins indicated that the MV-induced accumulation of RCD was limited to insoluble proteins with molecular masses of approximately 200, 120, 80, and 40 kDa. We conclude that MV results in a rapid onset of oxidative injury in the diaphragm and that insoluble proteins are primary targets of MV-induced protein oxidation. PMID- 12777409 TI - Altered functional coupling of coronary K+ channels in diabetic dyslipidemic pigs is prevented by exercise. AB - Chronic hyperglycemia and hypercholesterolemia have been shown to alter ionic currents in vascular smooth muscle. We tested the hypothesis that the combined effect of hyperglycemia and hyperlipidemia (diabetic dyslipidemia) would increase the Ca2+-sensitive K+ (KCa) current as a compensatory response to an increase in intracellular Ca2+ concentration. We also hypothesized that exercise training would prevent this elevation in KCa current. Miniature Yucatan swine were randomly assigned to five groups: control, standard pig chow (C, n = 6); hyperlipidemic, high-fat pig chow (H, n = 5); diabetic, standard pig chow (D, n = 7); diabetic, high-fat pig chow ("diabetic dyslipidemic," DD, n = 12); and exercise-trained DD (DDX, n = 9). High-fat chow consisted of standard minipig chow supplemented with cholesterol (2%) and coconut oil. Increased coronary vasoconstriction assessed in vivo and in vitro in DD was prevented by exercise. Patch-clamp experiments performed on right coronary artery smooth muscle cells resulted in greater K+ current densities in the H, D, and DD groups vs. the DDX group between -10 and 40 mV. In fura 2-loaded cells, current activated by caffeine-induced Ca2+ release was greater in H, D, and DD compared with C and DDX (P < 0.05), whereas intracellular Ca2+ concentration was not different across groups. Finally, there were no differences in the KCa or Kv channel protein content between groups. These data indicate that hyperglycemia, hyperlipidemia, and diabetic dyslipidemia lead to elevated whole cell K+ current and increased functional coupling of KCa and Ca2+ release. Endurance exercise prevented increased coupling of Ca2+ release to KCa channel activation in diabetic dyslipidemia. PMID- 12777410 TI - Variability in the interpolated twitch torque for maximal and submaximal voluntary contractions. AB - The superimposed twitch technique is frequently used to study the degree of motor unit activation during voluntary effort. This technique is one of the preferred methods to determine the activation deficit (AD) in normal, athletic, and patient populations. One of the limitations of the superimposed twitch technique is its variability under given contractile conditions. The objective of this research was to determine the source(s) of variability in the superimposed twitch force (STF) for repeat measurements. We hypothesized that the variability in the AD measurements may be caused by the timing of the twitch force relative to the onset of muscle activation, by force transients during the twitch application, by small variations in the actual force from the nominal target force, and by variations in the resting twitch force. Twenty-eight healthy subjects participated in this study. Sixteen of these subjects participated in a protocol involving contractions at 50% of their maximal voluntary contraction (MVC) effort, whereas the remaining 12 participated in a protocol involving contractions at 100% of their MVC. Doublet-twitch stimuli were superimposed onto the 50 and 100% effort knee extensor muscle contractions, and the resting twitch forces, voluntary knee extensor forces, and STFs were then measured. The mean resting twitch forces obtained before and after 8 s of 50% of MVC were the same. Similarly, the mean STFs determined at 1, 3, 5, and 7 s into the 50% MVC were the same. The variations in twitch force were significantly smaller after accounting for the actual force at twitch application than those calculated from the prescribed forces during the 50% MVC protocol (P < 0.05). Furthermore, the AD and the actual force showed statistically significant negative correlations for the 50% MVC tests. The interpolated twitch torque determined for the maximal effort contractions ranged from 1 to 70%. In contrast to the protocol at 50% of MVC, negative correlations were only observed in 5 of the 12 subjects during the 100% effort contractions. These results suggest that small variations in the actual force from the target force can account for the majority of the variations in the STFs for submaximal but not maximal effort contractions. For the maximal effort contractions, large variations in the STF exist due to undetermined causes. PMID- 12777412 TI - Seeing social position: visualizing class in life and death. PMID- 12777413 TI - On the adulteration of bread as a cause of rickets. 1857. PMID- 12777414 TI - Commentary: bread and alum, syphilis and sunlight: rickets in the nineteenth century. PMID- 12777415 TI - Commentary: John Snow and alum-induced rickets from adulterated London bread: an overlooked contribution to metabolic bone disease. PMID- 12777416 TI - Commentary: Snow on rickets. PMID- 12777418 TI - Mortality by education in German speaking Switzerland, 1990-1997: results from the Swiss National Cohort. AB - BACKGROUND: The aim of this paper is to show for the first time mortality differentials by level of education for Swiss men and women. This work is of interest to public health efforts in Switzerland as well as for co-operative international research into the determinants of socioeconomic differentials in health and mortality. METHODS: This study is based on a longitudinal data set from the Swiss National Cohort, currently incorporating a probabilistic record linkage of the 1990 Swiss census, and all subsequent deaths until the end of 1997. The study population covers all Swiss nationals aged >/=25 years living in German speaking Switzerland, with 19.7 million person-years and 296 929 deaths observed. Educational gradients were analysed using standardized mortality ratios, multiple logistic regression, and the Relative Index of Inequality (RII). RESULTS: There were sizeable gradients in mortality by education for all age groups and both sexes. The mortality odds ratio decreased by 7.2% (95% CI: 7.0 7.5%) per additional year of education for men, and by 6.0% (95% CI: 5.6-6.3%) for women. In men, we found a steady decrease of the gradient from 13.1% (95% CI: 11.9-14.4%) in the age group 25-39 to 4.5% (95% CI: 4.0-5.0%) in the age group >/=75 years. For women in the age groups under 65 the gradients were smaller; over the age of 40 there was no decrease with increasing age. These results were fairly insensitive to variations in the parameters of record linkage. CONCLUSIONS: Despite a comparatively low overall mortality, Swiss men in the 1990s show larger relative gradients in mortality by education than men in other European countries in the 1980s, with the possible exception of younger men in Italy. In Switzerland there is a sizeable potential for further increasing overall life expectancy by reducing the mortality of those with a lower educational level. The results presented contribute to a reliable assessment of socioeconomic mortality differentials in Europe. PMID- 12777419 TI - Commentary: explanations of the difference in mortality risk between different educational groups. PMID- 12777421 TI - Income and risk of ischaemic heart disease in men and women in a Nordic welfare country. AB - BACKGROUND: The inverse relation between ischaemic heart disease (IHD) and income is well known among men, but it remains to be clarified whether the relationship between social gradient and IHD is similar for men and women. The present study explores the associations between income and IHD in men and women in a Nordic country. METHODS: We used data from two prospective population studies conducted in Copenhagen. A total of 22 782 subjects, 54% women, with initial examination between 1964 and 1992 were followed until 1996 for hospital admission or death from IHD. We performed survival analyses, taking traditional cardiovascular risk factors into account, and estimated IHD-free life expectancy by household income in men and women. RESULTS: During follow-up, 1803 men and 1258 women experienced an event of IHD (21% fatal). The hazards by deciles of income showed a non-linear graded inverse effect of income, with a large group of middle-income in which income was not associated with risk of IHD. The hazard ratio for highest versus lowest deciles was 0.53 (95% CI: 0.44-0.65). The association was attenuated by adjustment for risk factors, but remained statistically significant. The associations were similar for both sexes. Median IHD-free life expectancy for low income versus high-income groups was reduced by 9.4 and 7.0 years in men and women, respectively. CONCLUSIONS: The effect of household income on risk of IHD was graded and similar for men and women. The difference between high and low income, regarding IHD-free life expectancy, was considerable. PMID- 12777420 TI - Neighbourhood deprivation and health: does it affect us all equally? AB - BACKGROUND: Neighbourhood socioeconomic status (SES) may affect rich and poor residents differentially. Two models are proposed. Model 1: living in a non deprived neighbourhood is better for health because better collective material and social resources are available. Model 2: being poor (rich) relative to the neighbourhood average is associated with worse (better) health because of the discrepancy between an individual's situation and those around them. METHODS: Individual data from the Whitehall II study covering health, SES, and perceived status were linked to census data on neighbourhood deprivation. RESULTS: Both individual and neighbourhood deprivation increased the risk of poor general and mental health. There was a suggestion that the effect of living in a deprived area was more marked for poorer individuals, although interactions were not statistically significant. Poor people in poor neighbourhoods reported more financial and neighbourhood problems and rated themselves lowest on the ladder of society. CONCLUSIONS: We found no evidence that personal poverty combined with affluent neighbourhood had negative health consequences. Rather, living in a deprived neighbourhood may have the most negative health effects on poorer individuals, possibly because they are more dependent on collective resources in the neighbourhood. PMID- 12777422 TI - Income inequality and ischaemic heart disease in Danish men and women. AB - BACKGROUND: It has been hypothesized that areas with an unequal income distribution are less likely to invest in health and more likely to have a social environment that influences the development of ischaemic heart disease (IHD) METHODS: We used pooled data from two cohort studies conducted in Copenhagen to analyse the association between area income inequality and first admission to hospital or death from IHD in women and men while controlling for individual income and other IHD risk factors. A total of 11 685 women and 10 036 men, with initial health examinations between 1964 and 1992, were followed for a median of 13.8 years. Information on median income share at parish and municipality levels was obtained from population registers. RESULTS: During follow-up 1700 men and 1204 women experienced an IHD event. At parish level income share was inversely associated with an increased risk of IHD in men (hazard ratio [HR](most versus least equal quartile) = 0.85 (95% CI: 0.73-0.98). Among women there was no relation between parish income inequality and IHD. Subject's household income was inversely related to IHD, and when this variable was controlled for, the association between income inequality at parish level and IHD in men attenuated slightly. When behavioural and biological risk factors were entered into the Cox model this relation attenuated further. However, some of these risk factors might mediate rather than confound the effect of income inequality. The association between income inequality at municipality level and IHD was insignificant for men, while in women the relation had a curved shape with those living in the least equal areas having the lowest risk. CONCLUSIONS: This study provides no clear evidence for an association between income inequality measured at parish or municipality level and IHD in Danish adults. The associations were weak and varied between different strata and geographical levels. PMID- 12777423 TI - Gender-related differences in the association between socioeconomic status and self-reported diabetes. AB - BACKGROUND: The prevalence of diabetes has been steadily increasing in Western countries. We investigated the impact of socioeconomic status (SES) on the prevalence of self-reported diabetes, and its differences between genders. METHODS: Data for this investigation were derived from the second cycle of the National Population Health Survey conducted in 1996-1997. A total of 39 021 subjects (17 730 males and 21 291 females) >/=40 years of age who answered the question about diabetes were included in the present analysis. Educational attainment and income adequacy were used as indicators of SES. Multiple logistic regression models were constructed for men and women separately to assess the effects of SES on the prevalence of diabetes after adjustment for age, area of residence, body mass index, and physical activity. RESULTS: and The prevalence of diabetes was 6.6% among men and 5.5% among women. The CONCLUSIONS: prevalence increased with decreasing income category and educational attainment in both genders. The odds ratios for income and education in relation to diabetes after adjustment remained significant in women, but attained unity in men. Canadian women >/=40 years of age of low SES have a relatively high prevalence of diabetes, independent of age, area of residence, obesity, and physical inactivity. PMID- 12777424 TI - Social inequalities in mortality in a retrospective cohort of civil servants in Barcelona. AB - BACKGROUND: The objective of this study is to describe the inequalities in mortality by occupational category and sex in a retrospective cohort of civil servants working in the city council of Barcelona (Spain). METHODS: The cohort was followed for the period 1984-1993. There were 11 647 men and 9001 women. Age adjusted hazard ratios (HR) of death for occupational categories and manual versus non-manual groups and 95% CI were derived from Cox proportional hazards models. RESULTS: For total deaths in males, compared with high-level professionals, auxiliary workers (HR = 1.30, 95% CI: 0.96-1.77), skilled manual workers (HR = 1.29, 95% CI: 0.95-1.77), unskilled manual workers (HR = 1.46, 95% CI: 1.07-1.98) and police and fire manual workers (HR = 1.42, 95% CI: 1.08-1.87) had higher risk of death. Among women, for all causes of mortality, only police manual workers had higher mortality (HR = 5.63, 95% CI: 1.89-16.7) whereas auxiliary workers had the lowest HR (HR = 0.51, 95% CI: 0.25-1.05). The HR comparing manual and non-manual categories for all causes of death was 1.29 for males (95% CI: 1.09-1.52) and 1.07 for females (95% CI: 0.77-1.49). Among males, whereas manual workers had lower cardiovascular mortality (HR = 0.85, 95% CI: 0.63-1.15), cancer mortality was higher in the manual category. No association between manual category and mortality was found among women. CONCLUSIONS: This study provides an analysis of social inequalities in mortality in a cohort from a Southern European urban area. PMID- 12777425 TI - Socioeconomic differentials in the temperature-mortality relationship in Sao Paulo, Brazil. AB - BACKGROUND: We investigated the impact of environmental temperature on mortality in Sao Paulo, Brazil, and examined differences in the temperature-mortality relationship with respect to cause, age group, and socioeconomic position (SEP). METHODS: Generalized additive Poisson regression models adjusted for non temperature related seasonal factors (including air pollution) were used to analyse daily mortality counts for selected causes from 1991 to 1994. Individuals were classified by the aggregate SEP of their area of residency. These were analysed as potential modifiers of the temperature-mortality relationship. RESULTS: Among the elderly we observed a 2.6% increase in all-cause mortality per degree increase in temperature above 20 degrees C, and a 5.5% increase per degree drop in temperature below 20 degrees C, after adjustment for confounding. Relationships were similar in children, but somewhat weaker in adults. Cold effects were present for deaths due to cardiovascular disease (CVD), respiratory, and other causes, with effects being greatest in the respiratory group. Heat effects were not found for CVD deaths in adults, but otherwise varied little by cause of mortality. There was little evidence for a modification of the mortality effects of cold or heat by SEP. CONCLUSIONS: These findings show that the U shaped pattern of the temperature-mortality relationship found in cooler northern countries occurs also in a sub-tropical city. In addition, the relative effects of temperature were similar in each socioeconomic grouping. PMID- 12777426 TI - Commentary: mortality from environmental factors, but which ones? PMID- 12777427 TI - Income, education, and blood pressure in adults in Jamaica, a middle-income developing country. AB - BACKGROUND: At present, little is known about how socioeconomic status (SES) is related to blood pressure (BP) and hypertension in developing countries. This cross-sectional study examined associations between SES and BP in 2082 adults from a peri-urban area of Jamaica, a middle-income developing country. METHODS: Hypertension (systolic BP >/=140 mmHg, diastolic BP >/=90 mmHg or current hypertensive medication use) was estimated based on self-reported medication use and the mean of the second and third of three manual BP measurements. Income and education were self-reported. Linear or logistic regressions were used to estimate multivariate associations between BP or hypertension and SES. RESULTS: Hypertension prevalence was 20% in men and 28% in women. In both men and women, the income distributions of BP and hypertension were non-linear, indicating elevated levels in low as well as in high-income groups. In contrast to the negative relationships typical for industrialized countries, multivariate adjusted BP and hypertension were highest in the wealthiest women. In men with some high school education, income was positively associated with BP, while there were negative associations in men with lesser education. Unlike women, mean BP were highest in poor men with limited education. Low SES men were also least likely to receive diagnosis and treatment. CONCLUSIONS: Socioeconomic status is related to BP and hypertension in Jamaica, although relationships are non-linear. Behavioural and environmental factors that explain elevated BP among both low and high SES adults in developing countries must be identified to develop effective prevention strategies. PMID- 12777428 TI - Commentary: epidemiological transition and socioeconomic inequalities in blood pressure in Jamaica. PMID- 12777429 TI - Child mortality, socioeconomic position, and one-parent families: independent associations and variation by age and cause of death. AB - BACKGROUND: Although the association between child mortality and socioeconomic status is well established, it is unclear whether child mortality differences by socioeconomic position are present at all ages. The association of one-parent families with mortality, and whether any such association is due to associated low socioeconomic position, is also not clear. METHODS: In all, 480 of 693 (69%) 0-14 year old deaths during 1991-1994 were linked to 1991 census records. Analyses were weighted to adjust for potential linkage bias. RESULTS: There was approximately twofold higher mortality among the lowest compared with the highest socioeconomic categories of education, income, car access, and neighbourhood deprivation. Occupational class differences were weaker. These socioeconomic differences in mortality were strongest among infants (particularly sudden infant death syndrome [SIDS] mortality), but similar across other age groups (1-4, 5-9, and 10-14 years). The socioeconomic differences were of a similar magnitude for unintentional injury, cancer, congenital, and other deaths. Multivariable analyses demonstrated persistent independent associations of education, income, car access, and neighbourhood deprivation with mortality. Rate ratios (adjusted for age and ethnicity) for one-parent families compared with two-parent or other families were 1.2 (95% CI: 1.0, 1.5) and 1.8 (95% CI: 1.2, 2.5) for all-cause and unintentional injury mortality, respectively. Further adjustment for socioeconomic factors reduced these associations to 0.8 (95% CI: 0.6, 1.2) and 1.2 (95% CI: 0.7, 2.2), respectively. CONCLUSIONS: There does not appear to be notable variation in relative risk terms of socioeconomic differences in child mortality by age or cause of death. Any association of one-parent families with child mortality is due to associated low socioeconomic position. PMID- 12777430 TI - The use of occupation and industry classifications in general population studies. AB - Occupation and industry classifications are used in epidemiological studies to classify study subjects according to their job and subsequently to study risk by job, to infer social class indicators, or to infer exposure to specific agents through job-exposure matrices. However, documentation on methodological aspects concerning the use of occupation and industry classifications is sparse within epidemiology. This paper reviews the diverse applications of occupation and industry classifications in population-based epidemiological studies. The different classifications in use are discussed, and criteria are given for choosing a classification in an epidemiological study. Finally, the reliability of coding for occupation and industry is reviewed. A further standardization of the use of occupation and industry classifications in epidemiology is recommended, in order to facilitate future comparisons between studies and fully exploit their possibilities, especially when occupational exposures are to be inferred. PMID- 12777431 TI - Commentary: standardized coding of occupational data in epidemiological studies. PMID- 12777432 TI - Effects of misclassification of causes of death on the power of a trial to assess the efficacy of a pneumococcal conjugate vaccine in The Gambia. AB - BACKGROUND: A reduction in cause-specific mortality may be the most important public health measure of the efficacy of a new vaccine. However, in developing countries, assignment of causes of deaths occurring outside hospitals can be assessed often only through the questioning of relatives about the signs and symptoms leading to death ('post-mortem questionnaire'). Causes assigned in this way have poor sensitivity and specificity. We illustrate the effects of this misclassification on the power of a large trial of a pneumococcal polysaccharide/protein conjugate vaccine with a mortality endpoint. METHODS: Required sample sizes to achieve a study with specified power were calculated for all-cause and acute lower respiratory tract infection (ALRI) mortality for different levels of sensitivity and specificity of post-mortem questionnaires. Data from active community-based surveillance and post-mortem questionnaires collected 1989-1993 from the study area were used in the calculations. FINDINGS: The mortality rate among children aged 6-29 months from all causes was 34.2 per 1000 child-years; 19% of deaths were attributable to ALRI. Assuming that pneumococci would be responsible for 50% of ALRI deaths and that the vaccine would cover 70% of disease serotypes and would be 90% effective against these serotypes, the expected efficacy of the vaccine would be 6.0% (19% x 50% x 70% x 90%) against all causes combined and 31.5% (50% x 70% x 90%) against deaths from ALRI. If, as suggested by various reports, the sensitivity and specificity of assigning a death to ALRI by post-mortem questionnaire are about 40% and 90% respectively, then the observed vaccine efficacy against ALRI (as classified using the post-mortem questionnaire) would fall to 20%, and the power to detect this would be reduced by approximately 40%. Furthermore, low sensitivity of diagnosis would lead to a falsely low estimate of the burden of ALRI mortality in the population and the trial might have greater power to detect a reduction in mortality from all causes combined than that estimated at the outset. CONCLUSIONS: Low sensitivity and specificity of diagnosis by post-mortem questionnaire may mean that the power of a trial to detect a reduction in all cause mortality is similar to that to detect a reduction in ALRI mortality. Since the latter is more susceptible to bias from misclassification of cause of death, all-cause mortality may be the most suitable endpoint. Similar considerations apply to trials of interventions against other diseases for which a cause specific endpoint is subject to substantial misclassification. PMID- 12777433 TI - The evolving pattern of avoidable mortality in Russia. AB - BACKGROUND: Life expectancy at birth in Russia is over 12 years less than in western Europe. This study explores the possible role of medical care in explaining this gap by examining the evolving pattern of mortality amenable to timely and effective medical care in Russia compared with Estonia, Latvia, and Lithuania, and the UK. METHODS: Analysis of standardized death rates from causes amenable to health care (treatable) or inter-sectoral health policies (preventable) in all regions and decomposition of differences in life expectancy between Russia and the UK by age, sex, and cause of death for the period 1965 1999/2000. RESULTS: Death rates from treatable causes remained stable between the mid-1960s and mid-1980s in Russia and the Baltic republics while steadily falling in the UK to less than half the rate in Russia. In the 1990s, rates increased in the former Soviet republics, reaching a peak in 1994 but reversing again in Russia in 1998. Deaths from causes amenable to inter-sectoral health interventions were higher in the UK in 1965 than in the Soviet Union but subsequently fell steadily while they increased in the East. Between 1965 and 1999, the male life expectancy gap between Russia and the UK rose from 3.6 to 15.1 years (women: 1.6 and 7.4 years). Treatable causes became an increasingly important contributor to this gap, accounting for almost 3 years by the end of the 1990s in men and 2 years in women. In Russia, elimination of treatable causes of death would have increased life expectancy by 2.9 years in men in 1995/99 compared with 1.2 years in the UK (women: 3.3 and 1.8 years), suggesting that, were the outcomes of health care achieved in the UK to be obtained in Russia, life expectancy for men might improve by about 1.7 years and for women by about 1.5 years. CONCLUSIONS: Our findings suggest that the Soviet health care system has failed to match the achievements of the West over the past three decades, highlighting the need to establish a system that provides effective and equitable care for the Russian population. PMID- 12777434 TI - Commentary: should we not go further than descriptions of avoidable mortality? PMID- 12777435 TI - Induced abortion and risk of subsequent miscarriage. AB - BACKGROUND: To evaluate the impact of surgically induced first-trimester abortion on the risk of miscarriage in a subsequent pregnancy. METHODS: The study is a pregnancy cohort study. It was conducted among 15 general hospitals or maternity and infant health institutes in Shanghai, China from November 1993 to March 1998. The abortion cohort consisted of pregnant women whose previous pregnancies were terminated by vacuum aspiration (98%). The reference cohort consisted of primigravidae. Subjects were recruited at 35-63 days of gestational age. A total of 2953 pregnant women were enrolled; 1502 in the abortion cohort, 1451 in the reference cohort. RESULTS: There were only 62 women lost to follow-up. The remaining 2891 women had 2732 live births, and 137 miscarriages. About 5.5% of pregnancies in the abortion cohort were miscarried and 4.0% in the reference cohort. Once potential confounders were controlled for by logistic regression, odds ratio (OR) of miscarriage between the abortion cohort and the reference cohort was 1.55 (95% CI: 1.08-2.23). The adjusted OR were 2.44 (95% CI: 1.16 5.15) among women who were recruited within 49 days of gestational age, and 1.72 (95% CI: 1.09-2.72) for the first-trimester miscarriage. CONCLUSIONS: Induced abortion by vacuum aspiration is associated with an increased risk of first trimester miscarriage in the subsequent pregnancy. PMID- 12777436 TI - Maternal serum oestrogen and androgen concentrations in preeclamptic and uncomplicated pregnancies. AB - BACKGROUND: Epidemiological studies show a substantially reduced risk of breast cancer in adult daughters of preeclamptic pregnancies, and modest risk reductions have been demonstrated for mothers also. Alterations in pregnancy hormone concentrations, particularly lower in utero exposure to oestrogen, are hypothesized to mediate this association. METHODS: Pregnancy hormone concentrations were measured in maternal sera collected at hospital admission for labour and delivery from 86 preeclamptic and 86 uncomplicated, singleton pregnancies matched on length of gestation, maternal age, parity, and type of delivery. RESULTS: Case and control pregnancies were similar in several maternal and pregnancy factors. Serum unconjugated oestradiol, oestrone, and oestriol concentrations were not lower in preeclamptic pregnancies in a matched analysis with adjustment for race and whether blood was collected before or after labour commenced. Serum unconjugated androstenedione (506.3 versus 316.0 ng/dl; P = 0.0007) and testosterone concentrations (214.5 versus 141.9 ng/dl; P = 0.004), however, were significantly higher in preeclamptic compared with control pregnancies, whereas dehydroepiandrosterone (DHEA) and DHEA sulphate did not differ. CONCLUSIONS: These data do not support the hypothesis that cancer risk in mothers and offspring of preeclamptic pregnancies is explained by exposure to lower maternal blood oestrogen concentrations, but raise the possibility that androgens play a role. PMID- 12777437 TI - Twelve-year changes in vascular risk factors and their associations with mortality in a cohort of 3499 Thais: the Electricity Generating Authority of Thailand Study. AB - BACKGROUND: Vascular mortality is increasing in economically developing countries such as Thailand but reliable data about the determinants of these changes are few. METHODS: In 1985, male and female employees of the Electricity Generating Authority of Thailand took part in a cardiovascular risk factor survey. In 1997, a follow-up survey was conducted and causes of death were determined for those subjects known to have died. Changes in levels of vascular risk factors over 12 years, and the associations of baseline risk factors with vascular mortality, were calculated. RESULTS: The 1985 survey recruited 3499 volunteers (average age 43 years) of whom 23% were female. In 1997, vital status was determined for 3318 (95%) and 2967 (85%) of the study participants were resurveyed. Mean levels of systolic blood pressure (SBP) and diastolic blood pressure (DBP), body mass index, total cholesterol and high density lipoprotein (HDL) cholesterol all increased over the 12-year follow-up period. Over the same time, the prevalence of diabetes also rose but the proportion of current smokers decreased. Vascular diseases were the most frequent cause of death during follow-up (n = 46), were positively associated with baseline age, SBP, DBP, smoking, diabetes, male sex, and total cholesterol, and were negatively associated with HDL cholesterol. CONCLUSIONS: Levels of most vascular risk factors worsened over the 12-year period between 1985 and 1997. The associations between baseline risk factor levels and vascular mortality were consistent with those observed in other populations. Interventions that control vascular risk factors have the potential to avert much premature vascular disease in Thailand. PMID- 12777438 TI - Commentary: cardiovascular implications of the epidemiological transition for the developing world: Thailand as a case in point. PMID- 12777439 TI - Periconceptional smoking and the male to female ratio in the offspring--re assessment of a recently proposed hypothesis. PMID- 12777442 TI - American Society of Clinical Oncology policy statement update: tobacco control- reducing cancer incidence and saving lives. 2003. AB - As an international medical society dedicated to cancer prevention, the American Society of Clinical Oncology (ASCO) advocates a fundamental reform of United States and international policy toward addictive tobacco products. ASCO's goal is the immediate reduction of tobacco use and ultimate achievement of a tobacco-free world. The centerpiece of ASCO's policy is the recommendation for an independent commission to study the tobacco problem in all of its dimensions: social, medical, legal, and economic (both domestically and globally). The commission membership should include broad-based representation and expertise on tobacco issues. In ASCO's view, tobacco control efforts to date have been less than successful because they are too fragmented and incremental, leaving many important issues unaddressed. A more comprehensive solution could flow from this study, including input from a variety of government agencies involved with public health, agriculture, First Amendment and other legal considerations, and international trade. The study, within defined time limits, should culminate in a report that outlines a strategy for achieving immediate reduction of tobacco use and ultimate achievement of a tobacco-free world, including explicit plans and a timetable for implementation. Although this comprehensive approach to tobacco control will take many years to implement even under the best of circumstances, there are certain measures that could be undertaken immediately with meaningful impact on tobacco usage. These include: Increasing efforts to discourage tobacco use, particularly among the young Raising federal excise taxes by at least $2 per pack and encouraging states to consider tobacco taxes as a first resort in revenue enhancement Ensuring that tobacco settlement funds be devoted only to health-related projects, including medical treatment, biomedical research, and tobacco prevention efforts Requiring disclosure of all ingredients in tobacco products Comprehensively reforming third-party payment for tobacco cessation efforts Additional restriction of secondhand smoke in any places where the public may congregate Supporting necessary research into tobacco addiction, toxicities, and prevention strategies Enhancing global tobacco control, including a halt of United States government promotion of tobacco products PMID- 12777443 TI - Maintenance of normal blood pressure and renal functions are independent effects of angiotensin-converting enzyme. AB - Angiotensin-converting enzyme (ACE) is expressed in many tissues, including vasculature and renal proximal tubules, and its genetic ablation in mice causes abnormal renal structure and functions, hypotension, and male sterility. To test the hypothesis that specific physiological functions of ACE are mediated by its expression in specific tissues, we generated different mouse strains, each expressing ACE in only one tissue. Here, we report the properties of two such strains of mice that express ACE either in vascular endothelial cells or in renal proximal tubules. Because of the natural cleavage secretion process, both groups also have ACE in the serum. Both groups were as healthy as wild-type mice, having normal kidney structure and fluid homeostasis, though males remained sterile, because they lack ACE expression in sperm. Despite equivalent serum ACE and angiotensin II levels and renal functions, only the group that expressed ACE in vascular endothelial cells had normal blood pressure. Expression of ACE, either in renal proximal tubules or in vasculature, is sufficient for maintaining normal kidney functions. However, for maintaining blood pressure, ACE must be expressed in vascular endothelial cells. These results also demonstrate that ACE-mediated blood pressure maintenance can be dissociated from its role in maintaining renal structure and functions. PMID- 12777446 TI - Sex differences in the modulation of K+ currents in diabetic rat cardiac myocytes. AB - A transient (Ipeak) and a sustained (Isus) outward K+ current were measured, using whole-cell voltage-clamp methods, in isolated rat ventricular myocytes obtained by enzymatic dispersion. A comparison was made between male and female rats following induction of (insulin-deficient) diabetes with streptozotocin (STZ). In control (non-diabetic) rats, both currents were smaller in cells obtained from females, as compared to males (P<0.005). However, whereas inducing diabetes in male rats significantly attenuated both Ipeak and Isus (P<0.005), Ipeak was unchanged in female diabetic rats. Isus was significantly (P<0.005) reduced, but the extent of reduction was smaller (P<0.02) than in males. The formation of angiotensin II (ATII) or endothelin-1 (ET-1) was blocked using inhibitors of angiotensin-converting enzyme (ACE) and endothelin-converting enzyme (ECE), respectively. In cells from diabetic males both inhibitors significantly (P<0.005) enhanced K+ currents. In contrast, no effect was observed in cells from female diabetic rats. However, in ovariectomized (Ovx) diabetic females the in vitro inhibition of ATII and ET-1 formation augmented the two K+ currents, but not when oestradiol was administered in vivo prior to cell isolation. In cells from diabetic males, incubation with 100 nM 17beta-oestradiol significantly (P<0.005) enhanced both Ipeak and Isus. This effect was blocked if ATII or ET-1 was added to the medium. These results show that autocrine modulation of K+ currents by renin-angiotensin and endothelin systems is attenuated or absent in female diabetic rats. Oestradiol plays a key role in reducing this modulation. These results may underlie some of the sex differences associated with development of cardiac arrhythmias. PMID- 12777449 TI - Effects of leg muscle tendon vibration on group Ia and group II reflex responses to stance perturbation in humans. AB - Stretching the soleus (Sol) muscle during sudden toe-up rotations of the supporting platform in a standing subject evokes a short-latency response (SLR) and a medium-latency response (MLR). The aim of the present investigation was to further explore the afferent and spinal pathways mediating the SLR and MLR in lower limb muscles by means of tendon vibration. In seven subjects, toe-up or toe down rotations were performed under: (1) control, (2) continuous bilateral vibration at 90 Hz of Achilles' tendon or tibialis anterior (TA) tendon, and (3) post-vibration conditions. Sol and TA background EMG activity and reflex responses were bilaterally recorded and analysed. Toe-up rotations induced SLRs and MLRs in Sol at average latencies of 40 and 66 ms, respectively. During vibration, the latency of both responses increased by about 2 ms. The area of the SLR significantly decreased during vibration, regardless of the underlying background activity, and almost returned to control value post-vibration. The area of Sol MLR was less influenced by vibration than SLR, the reduction being negligible with relatively high background activity. However, contrary to SLR, MLR was even more reduced post-vibration. Toe-down rotations induced no SLR in the TA, while a MLR was evoked at about 81 ms. The area of TA MLR decreased slightly during vibration but much more post-vibration. SLRs and MLRs were differently affected by changing the vibration frequency to 30 Hz: vibration had a negligible effect on the SLR, but still produced a significant effect on the MLR. The independence from the background EMG of the inhibitory effect of vibration upon the SLR suggests that vibration removes a constant amount of the Ia afferent input. This can be accounted for by either presynaptic inhibition of group Ia fibres or a 'busy-line' phenomenon. The differential effect of vibration on SLRs and MLRs is compatible with the notions that spindle primaries have a higher sensitivity to vibration than secondaries, and that group II afferent fibres are responsible for the production of the MLR. The decrease of MLRs but not SLRs after vibration is discussed in terms of an interaction between peripheral and central drive on group II interneurones in order to produce sufficient EMG activity to maintain a given postural set. PMID- 12777448 TI - Acidosis decreases low Ca(2+)-induced neuronal excitation by inhibiting the activity of calcium-sensing cation channels in cultured mouse hippocampal neurons. AB - The effects of extracellular pH (pHo) on calcium-sensing non-selective cation (csNSC) channels in cultured mouse hippocampal neurons were investigated using whole-cell voltage-clamp and current-clamp recordings. Decreasing extracellular Ca2+ concentrations ([Ca2+]o) activated slow and sustained inward currents through the csNSC channels. Decreasing pHo activated amiloride-sensitive transient proton-gated currents which decayed to baseline in several seconds. With proton-gated channels inactivated by pre-perfusion with low pH solution or blocked by amiloride, decreasing pHo to 6.5 inhibited the csNSC currents with a leftward shift of the Ca2+ dose-inhibition curve. Increasing pH to 8.5, on the other hand, caused a rightward shift of the Ca2+ dose-inhibition curve and potentiated the csNSC currents. Intracellular alkalinization following bath perfusion of quinine mimicked the potentiation of the csNSC currents by increasing pHo, while intracellular acidification by addition and subsequent withdrawal of NH4Cl mimicked the inhibition of the csNSC currents by decreasing pHo. Intracellular pH (pHi) imaging demonstrated that decreasing pHo induced a corresponding decrease in pHi. Including 30 mM Hepes in the pipette solution eliminated the effects of quinine and NH4Cl on the csNSC currents, but only partially reduced the effect of lowering pHo. In current-clamp recordings, decreasing [Ca2+]o induced sustained membrane depolarization and excitation of hippocampal neurons. Decreasing pHo to 6.5 inhibited the low [Ca2+]o-induced csNSC channel-mediated membrane depolarization and the excitation of neurons. Our results indicate that acidosis may inhibit low [Ca2+]o-induced neuronal excitation by inhibiting the activity of the csNSC channels. Both the extracellular and the intracellular sites are involved in the proton modulation of the csNSC channels. PMID- 12777450 TI - Having it both ways? Vasoconstriction in contracting muscles. PMID- 12777451 TI - Presynaptic rat Kv1.2 channels suppress synaptic terminal hyperexcitability following action potential invasion. AB - Voltage-gated K+ channels activating close to resting membrane potentials are widely expressed and differentially located in axons, presynaptic terminals and cell bodies. There is extensive evidence for localisation of Kv1 subunits at many central synaptic terminals but few clues to their presynaptic function. We have used the calyx of Held to investigate the role of presynaptic Kv1 channels in the rat by selectively blocking Kv1.1 and Kv1.2 containing channels with dendrotoxin K (DTX-K) and tityustoxin-Kalpha (TsTX-Kalpha) respectively. We show that Kv1.2 homomers are responsible for two-thirds of presynaptic low threshold current, whilst Kv1.1/Kv1.2 heteromers contribute the remaining current. These channels are located in the transition zone between the axon and synaptic terminal, contrasting with the high threshold K+ channel subunit Kv3.1 which is located on the synaptic terminal itself. Kv1 homomers were absent from bushy cell somata (from which the calyx axons arise); instead somatic low threshold channels consisted of heteromers containing Kv1.1, Kv1.2 and Kv1.6 subunits. Current-clamp recording from the calyx showed that each presynaptic action potential (AP) was followed by a depolarising after-potential (DAP) lasting around 50 ms. Kv1.1/Kv1.2 heteromers had little influence on terminal excitability, since DTX-K did not alter AP firing. However TsTX-Kalpha increased DAP amplitude, bringing the terminal closer to threshold for generating an additional AP. Paired pre- and postsynaptic recordings confirmed that this aberrant AP evoked an excitatory postsynaptic current (EPSC). We conclude that Kv1.2 channels have a general presynaptic function in suppressing terminal hyperexcitability during the depolarising after-potential. PMID- 12777454 TI - 90Y-ibritumomab tiuxetan in the treatment of relapsed or refractory B-cell non Hodgkin's lymphoma. AB - OBJECTIVE: Non-Hodgkin's lymphoma (NHL) is the most frequently diagnosed malignancy of the immune system, with more than 53,900 new cases diagnosed in 2002. Conventional cancer therapies cure many, but not the majority of, cases of the aggressive forms of NHL, and the more indolent and follicular forms of the disease that affect nearly half of all patients with NHL are considered incurable. In the absence of cure or survival benefits, treatments such as radioimmunotherapy that induce remission and prolong time off therapy are considered valuable. (90)Y-Ibritumomab tiuxetan recently became the first radioimmunotherapy agent to be approved for commercial use by the U.S. Food and Drug Administration. After reading this article, the nuclear medicine technologist should be able to understand the incidence and prevalence of NHL, describe the ibritumomab tiuxetan therapy protocol, explain specific infusion techniques for this protocol, list acquisition parameters after injection of (111)In-ibritumomab tiuxetan, and describe specific safety techniques to keep risk as low as reasonably achievable while performing the therapy protocol. PMID- 12777455 TI - Utility of posterior oblique views in (99m)Tc-DMSA renal scintigraphy in children. AB - OBJECTIVE: Procedure guidelines suggest that optimal (99m)Tc-dimercaptosuccinic acid (DMSA) planar scintigraphy of the kidney should include right and left posterior oblique views in addition to the posterior projection. However, in a small number of restless children, it is sometimes difficult to get 3 good quality images. The aim of this study was to evaluate the prevalence of cases in which posterior oblique views were useful for interpreting (99m)Tc-DMSA renal scintigraphy. METHODS: Three nuclear medicine specialists were asked to interpret 40 (99m)Tc-DMSA renal scans twice, first on the basis of the posterior projection only and then by using the posterior and the right and left posterior oblique views. RESULTS: The oblique posterior views were considered useful by observers 1 and 2 for 4 kidneys and by observer 3 for 5 kidneys and were considered somewhat useful for up to 7 kidneys. The addition of oblique posterior views changed the interpretation on 5 occasions for observer 1, on 9 occasions for the observer 3, and on no occasion for observer 2. On average, therefore, changes in interpretation occurred for fewer than 6% of the kidneys. Moreover, no relationship was observed between the opinion of the clinicians that oblique views were useful and changes in the scintigraphic interpretations. CONCLUSION: Oblique views were found useful in only a few cases and, even in these cases, did not significantly modify the interpretations. Therefore, when restless children are being imaged, the focus should be on obtaining a good posterior projection, even at the price of not having oblique posterior views. PMID- 12777456 TI - Testing different storage conditions for (99m)Tc-MAG3 kit: can hot fractioning reduce the cost per unit dose? AB - Since its release for routine clinical use, (99m)Tc-mercaptoacetyltriglycine (MAG3) has become an important alternative to (131)I-labeled orthoiodohippuran. The cold kit for MAG3 is expensive, especially in developing countries. Therefore, unique storage conditions should be provided for cost reduction. Cold fractioning is a well-known procedure but has special requirements, such as a nitrogen tank and a laminar flow hood. The aim of this study was to prolong the shelf life of (99m)Tc-labeled MAG3 by a hot fractioning method, which separates the patient doses after (99m)Tc labeling. The radiochemical purity of the (99m)Tc labeled MAG3 kit was tested under different storage conditions. Hot fractioning of the (99m)Tc-labeled MAG3 kit was found to be a possible alternative to cold fractioning for routine clinical studies. PMID- 12777457 TI - Severe systemic reaction to (99m)Tc-methylene diphosphonate: a case report. AB - We report an unusual severe systemic reaction that occurred in a woman after a (99m)Tc-methylene diphosphonate bone scan and for which no alternative explanation could be found. The bone scintigram showed diffusely increased uptake in the liver and kidneys accompanied by reversible dysfunction of these organs and dermatologic manifestations. We speculate that an immune-mediated mechanism may have caused this unusual reaction. PMID- 12777458 TI - Diffusely increased uptake by thoracic vertebrae on bone scintigraphy in midcourse of lung cancer irradiation: a case report. AB - Bone scintigraphy performed on a patient during the middle of radiation therapy for an inoperable left lung malignancy showed diffusely increased uptake in the thoracic vertebrae and relatively increased uptake in the ribs of the left thorax. This bone scan finding is apparently a transient phenomenon that occurs in response to irradiation and eventually leads to photon deficiency or photopenia of the vertebrae. However, this transiently increased uptake of the thoracic spine, compared with uptake in the lumbar spine, mimics diffusely decreased uptake or photopenia of the lumbar vertebrae and may be misinterpreted as an effect of irradiation of the abdominal region. In the case of asymmetric uptake between the thoracic and lumbar spine, a carefully taken history of the timing and location of irradiation is necessary to avoid misinterpretation. PMID- 12777459 TI - Salivary gland scintigraphy: the use of semiquantitative analysis for uptake and clearance. AB - OBJECTIVE: Quantitative analysis of (99m)Tc-pertechnetate salivary gland scintigraphy has been used in the evaluation of salivary gland function, but so far no one method can be considered optimal for this task. In this study, a semiquantitative method providing 2 functional parameters for objective assessment of salivary gland function by scintillation camera imaging was tested. METHODS: Twenty-one patients referred for (99m)Tc-pertechnetate thyroid scanning were studied. Two patients with salivary complaints were also included. Dynamic imaging of the anterior head using a scintillation camera was started after a bolus intravenous injection of 185 MBq (5 mCi) (99m)Tc-pertechnetate at 1 frame per 30 s for 30 min. At 15 min after injection, diluted lemon juice was administered orally. Analysis of the dynamic study included time-activity curves of 4 salivary glands (right and left parotid and right and left submandibular). Two parameters of function were defined: uptake rate, taken as the value of the initial slope of the time-activity curve, and washout fraction, which was the relative mobilizable radioactivity from each salivary gland after ingestion of the sialogogue. A parametric image of the washout fraction was also generated. RESULTS: The images showed gradual uptake in the parotid and submandibular glands. Washout was noted immediately after ingestion of the lemon juice. The pattern of the time-activity curve in all glands showed an early fast-rising part followed by a slow-rising component to nearly a plateau within 6-10 min after injection. The mean value of the uptake rate parameter was 0.10 +/- 0.09 cps/s. There was no significant difference between the parotid and submandibular glands or the right and left sides. Uptake in the parotid gland was 1.5-2 times that in the submandibular gland. The washout fraction was 1.40 +/- 1.60 for the parotid glands and 0.77 +/- 0.41 for the submandibular glands (P = 0.005). CONCLUSION: The quantitative analysis method including the uptake rate and the washout fraction parameters would enable objective assessment of salivary function and provide a reproducible means for follow-up of functional impairment in certain diseases. PMID- 12777460 TI - NMTCB 2003 task analysis report. AB - RATIONALE: The Nuclear Medicine Technology Certification Board (NMTCB) undertook a task analysis survey in the summer of 2002 as a part of its ongoing efforts to maintain the validity of its entry-level examination. METHODS: A task analysis survey, including sections on demographics, procedures, equipment, pharmaceuticals, and tasks performed or used by nuclear medicine technologists, was prepared and sent to 1,800 certified nuclear medicine technologists (CNMTs). Survey recipients were asked to indicate the frequency with which specific tasks are performed in their departments and whether these tasks are performed by nuclear medicine technologists or by other professionals. Criticality ratings for each task were determined by the NMTCB Board of Directors. These data were combined using the Kane weighting method to determine an importance rating for each task. Survey recipients were also asked which procedures are performed and which equipment and pharmaceuticals are used in nuclear medicine procedures in their institutions. CONCLUSION: A new task analysis for nuclear medicine technology is presented. It will form the basis for the NMTCB's entry-level examination, beginning in March 2004. Lists of procedures, equipment, and pharmaceuticals used in the practice of nuclear medicine technology are also presented. PMID- 12777464 TI - Metabolism of the unnatural anticancer lipid safingol, L-threo dihydrosphingosine, in cultured cells. AB - We studied the metabolism of radioactively labeled safingol (l-threo dihydrosphingosine) in primary cultured neurons, B104 neuroblastoma cells, and Swiss 3T3 fibroblasts, and compared it to that of its natural stereoisomer d erythro-dihydrosphingosine. Both sphingoid bases are used as biosynthetic precursors for complex sphingolipids, albeit to different rates. Whereas a considerable amount of the natural sphingoid base is also directed to the catabolic pathway (20-66%, cell type dependent), only a minor amount of the nonnatural safingol is subjected to catabolic cleavage, most of it being N acylated to the respective stereochemical variant of dihydroceramide. Interestingly, N-acylation of safingol to l-threo-dihydroceramide is less sensitive to fumonisin B1 than the formation of the natural d-erythro dihydroceramide. In addition, safingol-derived l-threo-dihydroceramide, unlike its physiologic counterpart, is not desaturated. Most of it either accumulates in the cells (up to 50%) or is used as a biosynthetic precursor of the respective dihydrosphingomyelin (up to 45%). About 5% is, however, glucosylated and channeled into the glycosphingolipid biosynthetic pathway. Our results demonstrate that, despite its nonnatural stereochemistry, safingol is recognized and metabolized preferentially by enzymes of the sphingolipid biosynthetic pathway. Furthermore, our data suggest that the cytotoxic potential of safingol is reduced rather than enhanced via its metabolic conversion. PMID- 12777465 TI - Cellular apoptosis is associated with increased caveolin-1 expression in macrophages. AB - Macrophage apoptosis is an important factor in determining the efficiency of the immune response, atherosclerotic lesion stability, and clearance of aged cells by phagocytosis. The involvement of caveolin-1 in the regulation of apoptosis has been previously suggested in fibroblasts and epithelial cells. Here we show that treatment of thioglycollate-elicited mouse peritoneal macrophages with various unrelated apoptotic agents, including simvastatin, camptothecin, or glucose deprivation, is associated with a specific and large increase in caveolin-1 expression. In contrast, caveolin-2 levels remain unaffected. Induction of apoptosis was measured by changes in cell morphology, annexin V-labeling, and DNA fragmentation. We demonstrate that caveolin-1 in macrophages is present in lipid rafts and colocalizes with phosphatidylserine (PS) at the cell surface of apoptotic macrophages. Our data suggest that caveolin-1 increase is an early event, closely accompanied by PS externalization and independent of caspase activation and nuclear DNA fragmentation. The increase in caveolin-1 levels does not require new protein synthesis, as cycloheximide does not prevent the apoptosis-mediated increase in caveolin-1 levels. We propose that increased levels of caveolin-1 characterize the apoptotic phenotype of macrophages. Caveolin-1 may be involved in the efficient externalization of PS at the surface of the apoptotic cells. PMID- 12777466 TI - Absorption and retinol equivalence of beta-carotene in humans is influenced by dietary vitamin A intake. AB - The effect of vitamin A supplements on metabolic behavior of an oral tracer dose of [14C]beta-carotene was investigated in a longitudinal test-retest design in two adults. For the test, each subject ingested 1 nmol of [14C]beta-carotene (100 nCi) in an emulsified olive oil-banana drink. Total urine and stool were collected for up to 30 days; concentration-time patterns of [14C]beta-carotene, [14C]retinyl esters, and [14C]retinol were determined for 46 days. On Day 53, the subjects were placed on a daily vitamin A supplement (10000 IU/day), and a second dose of [14C]beta-carotene (retest) was given on Day 74. All 14C determinations were made using accelerator mass spectrometry. In both subjects, the vitamin A supplementation was associated with three main effects: 1). increased apparent absorption: test versus retest values rose from 57% to 74% (Subject 1) and from 52% to 75% (Subject 2); 2). an approximately 10-fold reduction in urinary excretion; and 3). a lower ratio of labeled retinyl ester/beta-carotene concentrations in the absorptive phase. The molar vitamin A value of the dose for the test was 0.62 mol (Subject 1) and 0.54 mol (Subject 2) vitamin A to 1 mol beta-carotene. Respective values for the retest were 0.85 and 0.74. These results show that while less cleavage of beta-carotene occurred due to vitamin A supplementation, higher absorption resulted in larger molar vitamin A values. PMID- 12777467 TI - Sphingomyelinase D, a novel probe for cellular sphingomyelin: effects on cholesterol homeostasis in human skin fibroblasts. AB - Sphingomyelin (SM) and free cholesterol (FC) are concentrated in the plasma membranes of eukaryotes; however, the physiological significance of their association is unclear. A common tool for studying the role of membrane SM is digestion with bacterial sphingomyelinase (SMase) C, which hydrolyzes SM to ceramide. However, it is not known whether the observed effects of SMase C treatment are due to the loss of SM per se or to the signaling effects of ceramide. In this study, we tested SMase D from Corynebacterium pseudotuberculosis, which hydrolyzes SM to ceramide phosphate, as an alternative probe. This enzyme specifically hydrolyzed SM in fibroblasts without causing accumulation of ceramide. Treatment of fibroblasts with SMase D stimulated translocation of PM FC to intracellular sites by <20% of the rate observed after SMase C digestion. The cells regenerated SM nearly completely within 5 h after SMase C treatment. However, even after 20 h, no regeneration occurred following SMase D digestion. These findings suggest that the translocation of PM FC caused by SMase C digestion is due to the cellular effects of ceramide rather than the loss of SM. Since ceramide phosphate does not appear to have such effects, we suggest that SMase D is a useful probe of membrane SM. PMID- 12777468 TI - Endotoxin down-regulates ABCG5 and ABCG8 in mouse liver and ABCA1 and ABCG1 in J774 murine macrophages: differential role of LXR. AB - Several of the ATP binding cassette (ABC) transporters have recently been shown to play important roles in reverse cholesterol transport (RCT) and prevention of atherosclerosis. In the liver, ABCG5 and ABCG8 have been proposed to efflux sterols into the bile for excretion. ABCG5 and ABCG8 also limit absorption of dietary cholesterol and plant sterols in the intestine. In macrophages, ABCA1 and ABCG1 mediate cholesterol removal from these cells to HDL. Many of these ABC transporters are regulated by the liver X receptor (LXR). We have previously shown that endotoxin (lipopolysaccharide) down-regulates LXR in rodent liver. In the present study, we examined the in vivo and in vitro regulation of these ABC transporters by endotoxin. We found that endotoxin significantly decreased mRNA levels of ABCG5 and ABCG8 in the liver, but not in the small intestine. When endotoxin or cytokines (tumor necrosis factor and interleukin-1) were incubated with J774 murine macrophages, the mRNA levels of ABCA1 were decreased. This effect was rapid and sustained, and was associated with a reduction in ABCA1 protein levels. Endotoxin and cytokines also decreased ABCG1 mRNA levels in J774 cells. Although LXR is a positive regulator of ABCA1 and ABCG1, we did not observe a reduction in protein levels of LXR or in binding of nuclear proteins to an LXR response element in J774 cells. The decrease in ABCG5 and ABCG8 levels in the liver as well as a reduction in ABCA1 and ABCG1 in macrophages during the host response to infection and inflammation coupled with other previously described changes in the RCT pathway may aggravate atherosclerosis. PMID- 12777469 TI - GLP-1 stimulates glucose-derived de novo fatty acid synthesis and chain elongation during cell differentiation and insulin release. AB - Glucagon-like peptide-1 (GLP-1, 7-36) is capable of restoring normal glucose tolerance in aging, glucose-intolerant Wistar rats and is a potent causal factor in differentiation of human islet duodenal homeobox-1-expressing cells into insulin-releasing beta cells. Here we report stable isotope-based dynamic metabolic profiles of rat pancreatic epithelial (ARIP) and human ductal tumor (PANC-1) cells responding to 10 nM GLP-1 treatment in 48 h cultures. Macromolecule synthesis patterns and substrate flow measurements using gas chromatography/mass spectrometry (MS) and the stable [1,2-13C2]glucose isotope as the tracer showed that GLP-1 induced a significant 20% and 60% increase in de novo fatty acid palmitate synthesis in ARIP and PANC-1 cells, respectively, and it also induced a significant increase in palmitate chain elongation into stearate utilizing glucose as the primary substrate. Distribution of 13C in other metabolites indicated no changes in the rates of nucleic acid ribose synthesis, glutamate oxidation, or lactate production. Tandem high-performance liquid chromatography-ion trap MS analysis of the culture media demonstrated mass insulin secretion by GLP-1-treated tumor cells. Metabolic profile changes in response to GLP-1-induced cell differentiation include selective increases in de novo fatty acid synthesis from glucose and consequent chain elongation, allowing increased membrane formation and greater insulin availability and release. PMID- 12777471 TI - Determinants of low HDL levels in familial combined hyperlipidemia. AB - In familial combined hyperlipidemia (FCHL), affected family members frequently have reduced levels of HDL cholesterol, in addition to elevated levels of total cholesterol and/or triglycerides (TGs). In the present study, we focused on those determinants that are important regulators of HDL cholesterol levels in FCHL, and measured postheparin plasma activities of hepatic lipase (HL), lipoprotein lipase, cholesterol ester transfer protein, and phospholipid transfer protein (PLTP) in 228 subjects from 49 FCHL families. In affected family members (n = 88), the levels of HDL cholesterol, HDL2 cholesterol, HDL3 cholesterol, and apolipoprotein A-I were lower than in unaffected family members (n = 88) or spouses (n = 52). The main change was the reduction of HDL2 cholesterol by 25.4% in affected family members (P < 0.001 vs. unaffected family members; P = 0.003 vs. spouses). Affected family members had higher HL activity than unaffected family members (P = 0.001) or spouses (P = 0.013). PLTP activity was higher in affected than unaffected family members (P = 0.025). In univariate correlation analysis, a strong negative correlation was observed between HL activity and HDL2 cholesterol (r = -0.339, P < 0.001). Multivariate regression analysis demonstrated that gender, HL activity, TG, and body mass index have independent contributions to HDL2 cholesterol levels. We suggest that in FCHL, TG enrichment of HDL particles and enhanced HL activity lead to the reduction of HDL cholesterol and HDL2 cholesterol. PMID- 12777470 TI - Lipoprotein lipase and hepatic lipase: their relationship with HDL subspecies Lp(A-I) and Lp(A-I,A-II). AB - HDL subspecies Lp(A-I) and Lp(A-I,A-II) have different anti-atherogenic potentials. To determine the role of lipoprotein lipase (LPL) and hepatic lipase (HL) in regulating these particles, we measured these enzyme activities in 28 healthy subjects with well-controlled Type 1 diabetes, and studied their relationship with Lp(A-I) and Lp(A-I,A-II). LPL was positively correlated with the apolipoprotein A-I (apoA-I), cholesterol, and phospholipid mass in total Lp(A I), and with the apoA-I in large Lp(A-I) (r >or= 0.58, P >or= 0.001). HL was negatively correlated with all the above Lp(A-I) parameters plus Lp(A-I) triglyceride (r >or= -0.53, P or= 0.50, P 4-cholesten-3 one>7alpha-hydroxycholesterol>24-hydroxy-4-cholesten-3-one> cholesterol>25 hydroxy-4-cholesten-3-one>24-hydroxycholesterol>or=25-hydroxycholesterol. The possibility is discussed that the findings may have implications for oxysterol mediated regulation of gene expression. The very high activity of CYP27A1 towards the cholestanol precursor 4-cholesten-3-one may be of importance in connection with the accumulation of cholestanol in patients with CTX. PMID- 12777474 TI - Molecular characterization of the first avian LDL receptor: role in sterol metabolism of ovarian follicular cells. AB - Low levels of expression and sluggish sterol-mediated regulation have been likely reasons for the failure to molecularly characterize a bona fide LDL receptor (LDLR) in egg-laying species to date. The overall structure of the chicken LDLR, delineated here by cDNA cloning, has been conserved in evolution, since hallmark properties of mammalian LDLRs are already present in the avian protein. The chicken receptor appears to prefer LDL over VLDL as ligand, in compliance with its main role in providing lipoprotein-derived cholesterol for steroid production in ovarian follicular cells. This is also compatible with the fact that estrogen administration increased hepatic LDLR expression in roosters despite dramatically stimulated VLDL production. In cultured chicken embryo fibroblasts, expression of the receptor was induced by incubation with cholesterol synthesis inhibitors such as a statin. Furthermore, preincubation of induced cells with a specific anti receptor antibody blocks LDL endocytosis, demonstrating that the receptor is ligand-endocytosis competent. Finally, the distribution of LDLRs among the extraoocytic cell populations lends support to a three-cell model for estrogen production within the ovarian follicle. In summary, the molecular characterization of the first avian LDLR reveals novel information about evolutionary, structural, and functional aspects of members of the supergene family of LDLR-related proteins. PMID- 12777475 TI - Oxidized LDL activates phospholipase A2 to supply fatty acids required for cholesterol esterification. AB - We examined the roles of phospholipase A2 (PLA2) in oxidized LDL (oxLDL)-induced cholesteryl ester formation in macrophages. In [3H]oleic acid-labeled RAW264.7 cells and mouse peritoneal macrophages, oxLDL induced [3H]cholesteryl oleate formation with an increase in free [3H]oleic acid and a decrease in [3H]phosphatidylcholine. The changes in these lipids were suppressed by methyl arachidonyl fluorophosphonate (MAFP), a cytosolic PLA2 (cPLA2) inhibitor. However, MAFP had no effect on the ACAT activity or the binding and/or uptake of oxLDL. Stimulation with oxLDL in the presence of [3H]cholesterol increased [3H]cholesteryl ester bearing fatty acyl chains derived from cellular and/or exogenous (oxLDL) lipids. The formation of cholesteryl ester under this condition was also inhibited by MAFP, and the inhibitory effect was reversed by adding oleic acid. While oxLDL did not affect the activity or amounts of cPLA2, preincubation with oxLDL enhanced the release of oleic acid and arachidonic acid induced by ionomycin in RAW264.7 cells. 13(S)-hydroxyoctadecadienoic acid, but not 7-ketocholesterol, also enhanced ionomycin-induced oleic acid release. These results suggest that oxLDL induces cPLA2 activation, which contributes, at least in part, to the supply of fatty acids required for the cholesteryl esterification, probably through the acceleration by oxidized lipids of the catalytic action of cPLA2 in macrophages. PMID- 12777476 TI - Characterization of a novel mutation causing hepatic lipase deficiency among French Canadians. AB - Individuals with hepatic lipase (HL) deficiency are often characterized by elevated levels of triglycerides (TGs) and cholesterol. The aim of the present study was to characterize the molecular defect leading to severe HL deficiency in a Quebec-based kindred. In the proband and two of her brothers, the very low to undetectable HL activity resulted from compound heterozygosity for two rare HL gene mutations, a previously unknown missense mutation in exon 5 designated A174T and the previously reported T383M mutation in exon 8 of the HL gene. The mutation at codon 174 resulted in the substitution of alanine for threonine, a polar amino acid, in a highly conserved nonpolar region of the protein involved in the catalytic activity of the enzyme. The severe HL deficiency among the three related compound heterozygotes was associated with a marked TG enrichment of LDL and HDL particles. The two men with severe HL deficiency also presented with abdominal obesity, which appeared to amplify the impact of HL deficiency on plasma TG-rich lipoprotein levels. Our results demonstrated that HL deficiency in this Quebec kindred is associated with an abnormal lipoprotein-lipid profile, which may vary considerably in the presence of secondary factors such as abdominal obesity. PMID- 12777477 TI - A quick, reliable, and automated method for fat cell sizing. AB - Mean diameters of fat cells from abdominal tissues from 31 volunteers were determined by three methods based on fat cell isolation after collagenase digestion and methylene blue staining. The three methods were direct microscopy (Micro), manual measurement of diameters from digital images by using the public domain NIH Image program (Scion), and automated measurement of diameters from digital images using a customized program developed by Biomedical Imaging Resource at Mayo Clinic (AdCount). There was excellent agreement between the methods' measurement of mean abdominal fat cell diameter (concordance correlation coefficient >0.84). The Scion method gave slightly but systematically lower mean abdominal fat cell diameters than did either AdCount or Micro. The AdCount approach produced results that are comparable to those from Micro. Comparison of AdCount and Micro in measuring diameters of fat cells from thigh confirmed the good comparability between the two methods independent of fat depot. AdCount is very reliable, and the quickest and most objective of the three methods in measuring fat cell diameters from various depots. PMID- 12777478 TI - Arming the enemy: the evolution of resistance to self-proteins. AB - A remarkable range of novel antibiotics is attracting increasing interest as a major new weapon in the campaign against bacterial infection. They are based on the toxic peptides that provide the innate immune system of animals, and it is claimed that bacteria will be unable to evolve resistance to them because they attack the 'Achilles' heel' of bacterial membrane structure. Both experimental evidence and theoretical arguments suggest that this claim is doubtful. If so, the introduction of these substances into general use may provoke the evolution of resistance to our own defence proteins and thus compromise our natural defences against infection. PMID- 12777479 TI - Multiple effects on Clostridium perfringens binding, uptake and trafficking to lysosomes by inhibitors of macrophage phagocytosis receptors. AB - Clostridium perfringens is a Gram-positive, anaerobic bacterium that is the most common cause of gas gangrene (clostridial myonecrosis) in humans. C. perfringens produces a variety of extracellular toxins that are thought to be the major virulence factors of the organism. However, C. perfringens has recently been shown to have the ability to survive in a murine macrophage-like cell line, J774 33, even under aerobic conditions. In J774-33 cells, C. perfringens can escape the phagosome and gain access to the cytoplasm. Since the receptor that is used for phagocytosis can determine the fate of an intracellular bacterium, we used a variety of inhibitors of specific receptors to identify those used by J774-33 cells to phagocytose C. perfringens. It was found that the scavenger receptor and mannose receptor(s) were involved in the phagocytosis of C. perfringens. In the presence of complement, the complement receptor (CR3) was also involved in the binding and/or uptake of C. perfringens. Since the receptor inhibition studies indicated that the scavenger receptor played a major role in phagocytosis, C. perfringens binding studies were performed with a Chinese hamster ovary (CHO) cell line expressing the mouse SR-A receptor. The cell line expressing the SR-A receptor showed a significant increase in C. perfringens binding in comparison to the non-transfected CHO cells. In the absence of opsonizing antibodies, the Fc receptor was not used to phagocytose C. perfringens. Forcing the macrophages to use a specific receptor by using combinations of different receptor inhibitors led to only a slight increase in co-localization of intracellular C. perfringens with the late endosome-lysosome marker LAMP-1. Carbohydrate analysis of C. perfringens strain 13 extracellular polysaccharide confirmed the presence of mannose and negatively charged residues of glucuronic acid, which may provide the moieties that promote binding to the mannose and scavenger receptors, respectively. PMID- 12777480 TI - The Pix pilus adhesin of the uropathogenic Escherichia coli strain X2194 (O2 : K( ): H6) is related to Pap pili but exhibits a truncated regulatory region. AB - Adhesins provide a major advantage for uropathogenic Escherichia coli in establishing urinary tract infections (UTIs). A novel gene cluster responsible for the expression of a filamentous adhesin of the pyelonephritogenic E. coli strain X2194 has been identified, molecularly cloned, and characterized. The 'pix operon' contains eight open reading frames which exhibit significant sequence homology to corresponding genes in the pap operon encoding P pili, the prevalent E. coli adhesins in non-obstructive acute pyelonephritis in humans. Although a pixB gene corresponding to the PapB regulator was identified, a papI homologue could not be found in the pix operon. Instead, a fragment of the R6 gene of the highly uropathogenic E. coli strain CFT073 was identified upstream of pixB. The R6 gene is located in a pathogenicity island containing several pilus-encoding sequences and shows homology to a transposase of Chelatobacter heintzii. In a pixA-lacZ fusion system it was demonstrated that the expression of Pix pili is regulated at the transcriptional level by the R6 gene sequence. A significantly reduced transcription was observed by deleting this fragment and by lowering the growth temperature from 37 to 26 degrees C. In contrast to other filamentous adhesin systems, Pix pili are mainly expressed in the steady state growth phase and were not repressed by the addition of glucose. PMID- 12777481 TI - Lipoprotein signal peptidase of Streptococcus suis serotype 2. AB - This paper reports the complete coding sequence for a proliprotein signal peptidase (SP-ase) of Streptococcus suis, Lsp. This is believed to be the first SP-ase described for S. suis. SP-ase II is involved in the removal of the signal peptide from glyceride-modified prolipoproteins. By using in vitro transcription/translation systems, it was shown that the lsp gene was transcribed in vitro. Functionality of Lsp in Escherichia coli was demonstrated by using an in vitro globomycin resistance assay, to show that expression of Lsp in E. coli increased the globomycin resistance. An isogenic mutant of S. suis serotype 2 unable to produce Lsp was constructed and shown to process lipoproteins incorrectly, including an S. suis homologue of the pneumococcal PsaA lipoprotein. Five piglets were inoculated with a mixture of both strains in an experimental infection, to determine the virulence of the mutant strain relative to that of the wild-type strain in a competitive challenge experiment. The data showed that both strains were equally virulent, indicating that the knockout mutant of lsp is not attenuated in vivo. PMID- 12777482 TI - The Edwardsiella ictaluri O polysaccharide biosynthesis gene cluster and the role of O polysaccharide in resistance to normal catfish serum and catfish neutrophils. AB - Edwardsiella ictaluri, the causative agent of enteric septicaemia of catfish (ESC), expresses long O polysaccharide (OPS) chains on its surface. The authors previously reported the construction of an isogenic Ed. ictaluri OPS mutant strain and demonstrated that this strain is avirulent in channel catfish. This paper reports the cloning of the Ed. ictaluri OPS biosynthesis gene cluster and identification of the mutated gene in the OPS-negative strain. The sequenced region contains eight complete ORFs and one incomplete ORF encoding LPS biosynthesis enzymes. The mutated gene (designated wbiT) was similar to other bacterial galactose-4-epimerases. Glycosyl composition analysis indicated that wild-type Ed. ictaluri OPS contains higher amounts of galactose and N acetylgalactosamine than the OPS mutant strain, which correlated well with predicted functions of the genes identified in the OPS biosynthesis cluster. The OPS mutant had a relatively small, but significant, decrease in its ability to survive in normal catfish serum compared to wild-type Ed. ictaluri, but it retained the ability to resist killing by catfish neutrophils. PMID- 12777483 TI - The senX3-regX3 two-component regulatory system of Mycobacterium tuberculosis is required for virulence. AB - Two-component regulatory systems have been widely implicated in bacterial virulence. To investigate the role of one such system in Mycobacterium tuberculosis, a strain was constructed in which the senX3-regX3 system was deleted by homologous recombination. The mutant strain (Tame15) showed a growth defect after infection of macrophages and was attenuated in both immunodeficient and immunocompetent mice. Competitive hybridization of total RNA from the wild type and mutant strains to a whole-genome microarray was used to identify changes in gene expression resulting from the deletion. One operon was highly up regulated in the mutant, indicating that regX3 probably has a role as a repressor of this operon. Other genes which were up- or down-regulated were also identified. Many of the genes showing down-regulation are involved in normal growth of the bacterium, indicating that the mutant strain is subject to some type of growth slow-down or stress. Genes showing differential expression were further grouped according to their pattern of gene expression under other stress conditions. From this analysis 50 genes were identified which are the most likely to be controlled by RegX3. Most of these genes are of unknown function and no obvious motifs were found upstream of the genes identified. Thus, it has been demonstrated that the senX3-regX3 two-component system is involved in the virulence of M. tuberculosis and a number of genes controlled by this system have been identified. PMID- 12777484 TI - Structural and functional features of Rhodococcus ruber lipoarabinomannan. AB - The genus Rhodococcus is part of the phylogenetic group nocardioform actinomycetes, which also includes the genus MYCOBACTERIUM: Members of this phylogenetic group have a characteristic cell envelope structure, which is dominated by various complex lipids. Among these, lipoglycans are of particular interest since mycobacterial lipoarabinomannans are important immunomodulatory molecules that are likely to be involved in the subsequent fate of mycobacterial bacilli once inside phagocytic cells. Rhodococcus ruber is a species closely related to an established opportunistic human pathogen, Rhodococcus equi. This paper reports the isolation and characterization of R. ruber lipoarabinomannan, designated as RruLAM. SDS-PAGE and gas chromatography analyses revealed that RruLAM was of an intermediate size between Mycobacterium tuberculosis lipoarabinomannan and lipomannan. Using a combination of chemical degradation and (1)H, (13)C-NMR experiments, the carbohydrate structure of RruLAM was unambiguously shown to be composed of a linear (alpha1-->6)-Manp backbone substituted at some O-2 positions by a single t-alpha-Araf sugar unit. Integration of the anomeric proton signals provided an indication of the degree of branching as approximately 45 %. The RruLAM structure is much simpler than that established for M. tuberculosis lipoarabinomannan but is also different from that determined for the closely related species and opportunistic human pathogen, R. equi. RruLAM was unable to induce the production of TNF-alpha by either human or murine macrophage cell lines, suggesting that more sophisticated structures, such as phosphoinositol capping motifs, are required for such activity. PMID- 12777485 TI - The THI5 gene family of Saccharomyces cerevisiae: distribution of homologues among the hemiascomycetes and functional redundancy in the aerobic biosynthesis of thiamin from pyridoxine. AB - The THI5 gene family of Saccharomyces cerevisiae comprises four highly conserved members named THI5 (YFL058w), THI11 (YJR156c), THI12 (YNL332w) and THI13 (YDL244w). Each gene copy is located within the subtelomeric region of a different chromosome and all are homologues of the Schizosaccharomyces pombe nmt1 gene which is thought to function in the biosynthesis of hydroxymethylpyrimidine (HMP), a precursor of vitamin B(1), thiamin. A comprehensive phylogenetic study has shown that the existence of THI5 as a gene family is exclusive to those yeasts of the Saccharomyces sensu stricto subgroup. To determine the function and redundancy of each of the S. cerevisiae homologues, all combinations of the single, double, triple and quadruple deletion mutants were constructed using a PCR-mediated gene-disruption strategy. Phenotypic analyses of these mutant strains have shown the four genes to be functionally redundant in terms of HMP formation for thiamin biosynthesis; each promotes synthesis of HMP from the pyridoxine (vitamin B(6)) biosynthetic pathway. Furthermore, growth studies with the quadruple mutant strain support a previous proposal of an alternative HMP biosynthetic pathway that operates in yeast under anaerobic growth conditions. Comparative analysis of mRNA levels has revealed subtle differences in the regulation of the four genes, suggesting that they respond differently to nutrient limitation. PMID- 12777486 TI - The Candida albicans CTR1 gene encodes a functional copper transporter. AB - Copper and iron uptake in Saccharomyces cerevisiae are linked through a high affinity ferric/cupric-reductive uptake system. Evidence suggests that a similar system operates in Candida albicans. The authors have identified a C. albicans gene that is able to rescue a S. cerevisiae ctr1/ctr3-null mutant defective in high-affinity copper uptake. The 756 bp ORF, designated CaCTR1, encodes a 251 amino acid protein with a molecular mass of 27.8 kDa. Comparisons between the deduced amino acid sequence of the C. albicans Ctr1p and S. cerevisiae Ctr1p indicated that they share 39.6 % similarity and 33.0 % identity over their entire length. Within the predicted protein product of CaCTR1 there are putative transmembrane regions and sequences that resemble copper-binding motifs. The promoter region of CaCTR1 contains four sequences with significant identity to S. cerevisiae copper response elements. CaCTR1 is transcriptionally regulated in S. cerevisiae in response to copper availability by the copper-sensing transactivator Mac1p. Transcription of CaCTR1 in C. albicans is also regulated in a copper-responsive manner. This raises the possibility that CaCTR1 may be regulated in C. albicans by a Mac1p-like transactivator. A C. albicans ctr1-null mutant displays phenotypes consistent with the lack of copper uptake including growth defects in low-copper and low-iron conditions, a respiratory deficiency and sensitivity to oxidative stress. Furthermore, changes in morphology were observed in the C. albicans ctr1-null mutant. It is proposed that CaCTR1 facilitates transport of copper into the cell. PMID- 12777487 TI - A genetic system for the rapid isolation of aromatic-ring-hydroxylating dioxygenase activities. AB - Aromatic-ring-hydroxylating dioxygenases (ARHDOs) are key enzymes in the aerobic bacterial metabolism of aromatic compounds. They are of biotechnological importance as they function as biocatalysts in the stereospecific synthesis of chiral synthons and the degradation of aromatic pollutants. This report describes the development and validation of a system for the rapid isolation and characterization of specific ARHDO activities. The system is based on the identification of ARHDO gene segments that encode the enzymes' major functional determinants, on consensus primers for the direct amplification of such partial genes and on a 'recipient' ARHDO gene cluster for the insertion of the amplified segments. Previously, it has been shown that neither the N- nor the C-terminal portions but only the core region of the large or alpha-subunit of a class II ARHDO significantly influence substrate and product spectra. On the basis of these observations, consensus primers were designed for the amplification of the gene segment encoding the catalytic core of the large subunit. These primers were tested on 11 bacterial isolates known to metabolize aromatic compounds. In 10 cases, a gene fragment of expected length was amplified. DNA sequencing confirmed similarity to ARHDO alpha-subunit gene cores. The heterologously well-expressible bphA gene cluster of Burkholderia sp. strain LB400 was modified to facilitate the in-frame insertion of amplified segments. It was used successfully to express the resulting hybrid gene clusters and to form catalytically active chimaeric ARHDOs. The metabolic properties of these enzymes differed significantly from each other and from the parental ARHDO of strain LB400. These results indicate that the system described here can be used to rapidly isolate and functionally characterize ARHDO activities, starting from isolated strains, mixtures of organisms or samples of nucleic acids. Applications of the system range from the recruitment of novel ARHDO activities to an improved characterization of natural ARHDO diversity. PMID- 12777488 TI - Cloning and characterization of the groE heat-shock operon of the marine bacterium Vibrio harveyi. AB - The DNA region of the Vibrio harveyi chromosome containing the heat-shock genes groES and groEL was cloned, and the genes were sequenced. These genes are arranged in the chromosome in the order groES-groEL. Northern hybridization experiments with RNA from V. harveyi and a DNA probe carrying both groES and groEL genes showed a single, heat-inducible transcript of approximately 2200 nt, indicating that these genes form an operon. Primer extension analysis revealed a strong, heat-inducible transcription start site 59 nt upstream of groES, preceded by a sequence typical for the Escherichia coli heat-shock promoters recognized by the sigma(32) factor, and a weak transcription start site 25 nt upstream the groES gene, preceded by a sequence typical for sigma(70) promoters. Transcription from the latter promoter occurred only at low temperatures. The V. harveyi groE operon cloned in a plasmid in E. coli cells was transcribed in a sigma(32) dependent manner; the transcript size and the sigma(32)-dependent transcription start site were as in V. harveyi cells. Comparison of V. harveyi groE transcription regulation with the other well-characterized groE operons of the gamma subdivision of proteobacteria (those of E. coli and Pseudomonas aeruginosa) indicates a high conservation of the transcriptional regulatory elements among these bacteria, with two promoters, sigma(32) and sigma(70), involved in the regulation. The ability of the cloned groESL genes to complement E. coli groE mutants was tested: V. harveyi groES restored a thermoresistant phenotype to groES bacteria and enabled lambda phage to grow in the mutant cells. V. harveyi groEL did not abolish thermosensitivity of groEL bacteria but it complemented the groEL mutant with respect to growth of lambda phage. The results suggest that the GroEL chaperone may be more species-specific than the GroES co-chaperone. PMID- 12777489 TI - Intra-chromosomal heterogeneity between the four 16S rRNA gene copies in the genus Veillonella: implications for phylogeny and taxonomy. AB - Among the seven species characterized within the genus Veillonella, three (Veillonella dispar, Veillonella parvula and Veillonella atypica) have so far been isolated from human flora and during infectious processes. Sequencing and analysis of 16S rDNA (rrs) has been described as the best method for identification of Veillonella strains at the species level since phenotypic characteristics are unable to differentiate between species. rrs sequencing for the three species isolated from humans showed more than 98 % identity between them. Four rrs copies were found in the reference strains and in all the clinical isolates studied. The sequences of each rrs were determined for the clinical strain ADV 360.1, and they showed a relatively high level of heterogeneity (1.43 %). In the majority of cases, polymorphic positions corresponded to nucleotides allowing differentiation between the three species isolated from humans. Moreover, variability observed between rrs copies was higher than that between 16S rDNA sequences of V. parvula and V. dispar. Phylogenetic analysis showed that polymorphism between rrs copies affected the position of strain ADV 360.1 in the tree. Variable positions occurred in stems and loops belonging to variable and hypervariable regions of the 16S rRNA secondary structure but did not change the overall structure of the 16S rRNA. PCR-RFLP experiments performed on 27 clinical isolates of Veillonella sp. suggested that inter-rrs heterogeneity occurs widely among the members of the genus VEILLONELLA: These results, together with the lack of phenotypic criteria for species differentiation, give preliminary arguments for unification of V. dispar and V. parvula. PMID- 12777490 TI - Transposition in Lactobacillus delbrueckii subsp. bulgaricus: identification of two thermosensitive replicons and two functional insertion sequences. AB - In this report, it is shown that the rolling circle replicon pG(+)host and the theta replicon pIP501 are thermosensitive in Lactobacillus delbrueckii subsp. bulgaricus (Lactobacillus bulgaricus). Using a pIP501 derivative as a delivery vector for six insertion sequences originating from lactic acid bacteria, it is shown that IS1223 and IS1201 transpose in L. bulgaricus. PMID- 12777491 TI - Response of a strict anaerobe to oxygen: survival strategies in Desulfovibrio gigas. AB - The biochemical response to oxygen of the strictly anaerobic sulfate-reducing bacterium Desulfovibrio gigas was studied with the goal of elucidating survival strategies in oxic environments. Cultures of D. gigas on medium containing lactate and sulfate were exposed to oxygen (concentration 5-120 micro M). Growth was fully inhibited by oxygen, but the cultures resumed growth as soon as they were shifted back to anoxic conditions. Following 24 h exposure to oxygen the growth rate was as high as 70 % of the growth rates observed before oxygenation. Catalase levels and activity were enhanced by exposure to oxygen whereas superoxide-scavenging and glutathione reductase activities were not affected. The general pattern of cellular proteins as analysed by two-dimensional electrophoresis was altered in the presence of oxygen, the levels of approximately 12 % of the detected proteins being markedly increased. Among the induced proteins, a homologue of a 60 kDa eukaryotic heat-shock protein (Hsp60) was identified by immunoassay analysis. In the absence of external substrates, the steady-state levels of nucleoside triphosphates detected by in vivo (31)P-NMR under saturating concentrations of oxygen were 20 % higher than under anoxic conditions. The higher energy levels developed under oxygen correlated with a lower rate of substrate (glycogen) mobilization, but no experimental evidence for a contribution from oxidative phosphorylation was found. The hypothesis that oxygen interferes with ATP dissipation processes is discussed. PMID- 12777492 TI - Changes in GE2270 antibiotic production in Planobispora rosea through modulation of methylation metabolism. AB - Thiazolylpeptide GE2270 is a potent antibiotic inhibiting protein synthesis in Gram-positive bacteria. It is produced as a complex of 10 related metabolites, differing mainly in the degree of methylation, by fermentation of the rare actinomycete Planobispora rosea ATCC 53773. Addition of vitamin B12 to the fermentation medium doubled total complex production and markedly changed the relative production of the various GE2270 metabolites, enhancing the biosynthesis of the more methylated component A. Among methylation inhibitors, the addition of sinefungin increased the amount of factor D2, which differs from component A in the lack of a methyl group. Since sinefungin is an S-adenosyl-L-methionine methyltransferase-specific inhibitor, these results indicate that the methylation step converting D2 into A involves an S-adenosyl-L-methionine methyltransferase. Simultaneous supplementation of vitamin B12 and sinefungin led to a twofold increase in D2 concentration, showing that vitamin B12, in addition to having an effect on the late methylation step, exerts a stimulating action on antibiotic backbone synthesis. This is possibly due to its role in an unusual pathway of serine synthesis peculiar to P. rosea metabolism. Finally, fermentation medium modifications were shown to be useful for the production of industrially valuable levels of components A or D2 in the GE2270 complex as starting points for the production of new interesting semi-synthetic antibiotics. PMID- 12777493 TI - Characterization of the expression and activity of the periplasmic nitrate reductase of Paracoccus pantotrophus in chemostat cultures. AB - The periplasmic nitrate reductase (Nap) from Paracoccus pantotrophus has a role in cellular redox balancing. Previously, transcription from the nap promoter in P. pantotrophus was shown to be responsive to the oxidation state of the carbon substrate. During batch culture, expression was higher during growth on reduced substrates such as butyrate compared to more oxidized substrates such as succinate. In the present study the effect of growth rate on nap expression in succinate-, acetate- and butyrate-limited chemostat cultures was investigated. In all three cases transcription from the nap promoter and Nap enzyme activity showed a strong correlation. At the fastest growth rates tested for the three substrates nap expression and Nap activity were highest when growth occurred on the most reduced substrate (butyrate > acetate > succinate). However, in all three cases a bell-shaped pattern of expression was observed as a function of growth rate, with the highest levels of nap expression and Nap activity being observed at intermediate growth rates. This effect was most pronounced on succinate, where an approximately fivefold variation was observed, and at intermediate dilution rates nap expression and Nap activity were comparable on all three carbon substrates. Analysis of mRNA prepared from the succinate-grown cultures revealed that different transcription initiation start sites for the nap operon were utilized as the growth rate changed. This study establishes a new regulatory feature of nap expression in P. pantotrophus that occurs at the level of transcription in response to growth rate in carbon-limited cultures. PMID- 12777494 TI - AhlD, an N-acylhomoserine lactonase in Arthrobacter sp., and predicted homologues in other bacteria. AB - Quorum sensing is a signalling mechanism that controls diverse biological functions, including virulence, via N-acylhomoserine lactone (AHL) signal molecules in Gram-negative bacteria. With the aim of isolating strains or enzymes capable of blocking quorum sensing by inactivating AHL, bacteria were screened for AHL degradation by their ability to utilize N-3-oxohexanoyl-L-homoserine lactone (OHHL) as the sole carbon source. Among four isolates, strain IBN110, identified as Arthrobacter sp., was found to grow rapidly on OHHL, and to degrade various AHLs with different lengths and acyl side-chain substitutions. Co-culture of Arthrobacter sp. IBN110 and the plant pathogen Erwinia carotovora significantly reduced both the AHL amount and pectate lyase activity in co culture medium, suggesting the possibility of applying Arthrobacter sp. IBN110 in the control of AHL-producing pathogenic bacteria. The ahlD gene from Arthrobacter sp. IBN110 encoding the enzyme catalysing AHL degradation was cloned, and found to encode a protein of 273 amino acids. A mass spectrometry analysis showed that AhlD probably hydrolyses the lactone ring of N-3-hexanoyl-L-homoserine lactone, indicating that AhlD is an N-acylhomoserine lactonase (AHLase). A comparison of AhlD with other known AHL-degrading enzymes, Bacillus sp. 240B1 AiiA, a Bacillus thuringiensis subsp. kyushuensis AiiA homologue and Agrobacterium tumefaciens AttM, revealed 25, 26 and 21 % overall identities, respectively, in the deduced amino acid sequences. Although these identities were relatively low, the HXDH approximately H approximately D motif was conserved in all the AHLases, suggesting that this motif is essential for AHLase activity. From a genome database search based on the conserved motif, putative AhlD-like lactonase genes were found in several other bacteria, and AHL-degrading activities were observed in Klebsiella pneumoniae and Bacillus stearothermophilus. Furthermore, it was verified that ahlK, an ahlD homologue, encodes an AHL-degrading enzyme in K. pneumoniae. Accordingly, the current results suggest the possibility that AhlD like AHLases could exist in many other micro-organisms. PMID- 12777495 TI - The essential role of fumarate reductase in haem-dependent growth stimulation of Bacteroides fragilis. AB - Haem is required for optimal growth of the bacterial anaerobe Bacteroides fragilis. Previous studies have shown that growth in the presence of haem is coincident with increased yields of ATP from glucose, expression of b-type cytochromes and expression of fumarate reductase activity. This paper describes the identification of the genes that encode the cytochrome, iron-sulfur cluster protein and flavoprotein of the B. fragilis fumarate reductase. These genes, frdC, frdA and frdB, respectively, are organized in an operon. Nonpolar, in-frame deletions of frdC and frdB were constructed in the B. fragilis chromosome. These mutant strains had no detectable fumarate reductase or succinate dehydrogenase activity. In addition, the frd mutant strains showed a threefold increase in generation time, relative to the wild-type strain. Growth of these mutant strains was fully restored to the wild-type rate by the introduction of a B. fragilis replicon containing the entire frd operon. Growth of the frd mutant strains was partially restored by supplementing the growth medium with succinate, indicating that the frd gene products function as a fumarate reductase. During growth on glucose, the frd mutant strains showed a threefold decrease in cell mass yield, relative to the wild-type strain. These data indicate that fumarate reductase is important for both energy metabolism and succinate biosynthesis in B. fragilis. PMID- 12777496 TI - Site-directed mutagenesis of an extradiol dioxygenase involved in tetralin biodegradation identifies residues important for activity or substrate specificity. AB - The sequence of the extradiol dioxygenase ThnC, involved in tetralin biodegradation, was aligned with other extradiol dioxygenases involved in biodegradation of polycyclic compounds, and a three-dimensional model of ThnC, based on the structure of the previously crystallized 2,3-dihydroxybiphenyl dioxygenase from Burkholderia fungorum LB400, was built. In order to assess the functional importance of some non-active-site residues whose relevance could not be established by structural information, a number of positions surrounding the substrate-binding site were mutated in ThnC. Ten mutant proteins were purified and their activity towards 1,2-dihydroxytetralin, 1,2-dihydroxynaphthalene and 2,3-dihydroxybiphenyl was characterized. N213H, Q198H, G206M, A282R and A282G mutants increased k(cat)/K(m) at least twofold using 1,2-dihydroxytetralin as the substrate, thus showing that activity of ThnC is not maximized for this substrate. N213H and Q198H mutants increased k(cat)/K(m) using any of the substrates tested, thus showing the relevance for activity of these two histidines, which are highly conserved in dihydroxybiphenyl dioxygenases, but not present in dihydroxynaphthalene dioxygenases. Different substitutions in position 282 had different effects on general activity or substrate specificity, thus showing the functional importance of the most C-terminal beta-sheet of the protein. A251M and G206M mutants showed increased activity specifically for a particular substrate. N213H, G206M, A282R, A282G and Y177I substitutions resulted in enzymes more tolerant to acidic pH, the most striking effect being observed in mutant Y177I, which showed maximal activity at pH 5.5. In addition, Q198D and V175D mutants, which had altered K(m), also showed altered sensitivity to substrate inhibition, thus indicating that inhibition is exerted through the same binding site. This mutational analysis, therefore, identified conserved residues important for activity or substrate specificity, and also shed some light on the mechanism of substrate inhibition exhibited by extradiol dioxygenases. PMID- 12777497 TI - A single V317A or V317M substitution in Enzyme II of a newly identified beta glucoside phosphotransferase and utilization system of Corynebacterium glutamicum R extends its specificity towards cellobiose. AB - A catabolic system involved in the utilization of beta-glucosides in Corynebacterium glutamicum R and its spontaneous mutant variants allowing uptake of cellobiose were investigated. The system comprises a beta-glucoside-specific Enzyme IIBCA component (gene bglF) of the phosphotransferase system (PTS), a phospho-beta-glucosidase (bglA) and an antiterminator protein (bglG) from the BglG/SacY family of transcription regulators. The results suggest that transcription antitermination is involved in control of induction and carbon catabolite repression of bgl genes, which presumably form an operon. Functional analysis of the bglF and bglA products revealed that they are simultaneously required for uptake, phosphorylation and breakdown of methyl beta-glucoside, salicin and arbutin. Although cellobiose is not normally a substrate for BglF permease and is not utilized by C. glutamicum R, cellobiose-utilizing mutants can be obtained. The mutation responsible was mapped to the bgl locus and sequenced, and point mutations were found in codon 317 of bglF. These led to substitutions V317A and/or V317M near the putative PTS active-site H313 in the membrane spanning IIC domain of BglF and allowed BglF to act on cellobiose. Such results strengthen the evidence that the IIC domains can be regarded as selectivity filters of the PTS. PMID- 12777498 TI - Genetic and biochemical characterization of PrtA, an RTX-like metalloprotease from Photorhabdus. AB - Proteases play a key role in the interaction between pathogens and their hosts. The bacterial entomopathogen Photorhabdus lives in symbiosis with nematodes that invade insects. Following entry into the insect, the bacteria are released from the nematode gut into the open blood system of the insect. Here they secrete factors which kill the host and also convert the host tissues into food for the replicating bacteria and nematodes. One of the secreted proteins is PrtA, which is shown here to be a repeats-in-toxin (RTX) alkaline zinc metalloprotease. PrtA has high affinity for artificial substrates such as casein and gelatin and can be inhibited by zinc metalloprotease inhibitors. The metalloprotease also shows a calcium- and temperature-dependent autolysis. The prtA gene carries the characteristic RTX repeated motifs and predicts high similarity to proteases from Erwinia chrysanthemi, Pseudomonas aeruginosa and Serratia marcescens. The prtA gene resides in a locus encoding both the protease ABC transporter (prtBCD) and an intervening ORF encoding a protease inhibitor (inh). PrtA activity is detectable 24 h after artificial bacterial infection of an insect, suggesting that the protease may play a key role in degrading insect tissues rather than in overcoming the insect immune system. Purified PrtA also shows cytotoxicity to mammalian cell cultures, supporting its proposed role in bioconversion of the insect cadaver into food for bacterial and nematode development. PMID- 12777499 TI - Conditional expression of Mycobacterium smegmatis ftsZ, an essential cell division gene. AB - To understand the role of Mycobacterium smegmatis ftsZ (ftsZ(smeg)) in the cell division process, the ftsZ gene was characterized at the genetic level. This study shows that ftsZ(smeg) is an essential gene in that it can only be disrupted in a merodiploid background carrying another functional copy. Expression of ftsZ(smeg) in M. smegmatis from a constitutively active mycobacterial promoter resulted in lethality whereas that from a chemically inducible acetamidase (ami) promoter led to FtsZ accumulation, filamentation and cell lysis. To further understand the roles of ftsZ in cell division a conditionally complementing ftsZ(smeg) mutant strain was constructed in which ftsZ expression is controlled by acetamide. Growth in the presence of 0.2 % acetamide increased FtsZ levels approximately 1.4-fold, but did not decrease viability or change cell length. Withdrawal of acetamide reduced FtsZ levels, decreased viability, increased cell length and eventually lysed the cells. Finally, it is shown that ftsZ(smeg) function in M. smegmatis can be replaced with the Mycobacterium tuberculosis counterpart, indicating that heterologous FtsZ(tb) can independently initiate the formation of Z-rings and catalyse the septation process. It is concluded that optimal levels of M. smegmatis FtsZ are required to sustain cell division and that the cell division initiation mechanisms are similar in mycobacteria. PMID- 12777500 TI - GS-Aligner: a novel tool for aligning genomic sequences using bit-level operations. AB - A novel algorithm, GS-Aligner, that uses bit-level operations was developed for aligning genomic sequences. GS-Aligner is efficient in terms of both time and space for aligning two very long genomic sequences and for identifying genomic rearrangements such as translocations and inversions. It is suitable for aligning fairly divergent sequences such as human and mouse genomic sequences. It consists of several efficient components: bit-level coding, search for matching segments between the two sequences as alignment anchors, longest increasing subsequence (LIS), and optimal local alignment. Efforts have been made to reduce the execution time of the program to make it truly practical for aligning very long sequences. Empirical tests suggest that for relatively divergent sequences such as sequences from different mammalian orders or from a mammal and a nonmammalian vertebrate GS-Aligner performs better than existing methods. The program and data can be downloaded from http://pondside.uchicago.edu/~lilab/ and http://webcollab.iis.sinica.edu.tw/~biocom. PMID- 12777501 TI - The evolution of transcriptional regulation in eukaryotes. AB - Gene expression is central to the genotype-phenotype relationship in all organisms, and it is an important component of the genetic basis for evolutionary change in diverse aspects of phenotype. However, the evolution of transcriptional regulation remains understudied and poorly understood. Here we review the evolutionary dynamics of promoter, or cis-regulatory, sequences and the evolutionary mechanisms that shape them. Existing evidence indicates that populations harbor extensive genetic variation in promoter sequences, that a substantial fraction of this variation has consequences for both biochemical and organismal phenotype, and that some of this functional variation is sorted by selection. As with protein-coding sequences, rates and patterns of promoter sequence evolution differ considerably among loci and among clades for reasons that are not well understood. Studying the evolution of transcriptional regulation poses empirical and conceptual challenges beyond those typically encountered in analyses of coding sequence evolution: promoter organization is much less regular than that of coding sequences, and sequences required for the transcription of each locus reside at multiple other loci in the genome. Because of the strong context-dependence of transcriptional regulation, sequence inspection alone provides limited information about promoter function. Understanding the functional consequences of sequence differences among promoters generally requires biochemical and in vivo functional assays. Despite these challenges, important insights have already been gained into the evolution of transcriptional regulation, and the pace of discovery is accelerating. PMID- 12777502 TI - R5 retrotransposons insert into a family of infrequently transcribed 28S rRNA genes of planaria. AB - A small (100 bp) region of the 28S rRNA gene has been shown to serve as the target site for the insertion of non-long terminal repeat (non-LTR) retrotransposons in both arthropods and nematodes. Here we characterize a lineage of non-LTR retrotransposons that inserts into this target site in the phylum Platyhelminthes. Dugesiid planaria contain elements, named R5, that insert 8 bp upstream of the target site used by arthropod R2 elements. The complete sequence of this element from Girardia tigrina revealed that it encoded two open reading frames (ORFs). The second ORF contained reverse transcriptase and restriction enzyme-like endonuclease domains similar to those found in R2 and R4, the elements that insert into the 28S genes of nematodes. The closest relative of R5, however, was the element NeSL-1, which inserts into the spliced leader 1 exons of nematodes. The rRNA genes of dugesiid planaria are unusual in that they comprise two types of rDNA units that differ by 8%-10% in nucleotide sequence of the 18S and 28S coding regions. Type II units are transcribed in adult tissues at levels that are less than 1% that of the type I units. R5 elements were only found inserted in the type II units, where presumably they cause less harm to the host. A second unusual aspect of the dugesiid rRNA genes is that the target site for the R5 insertion is duplicated 300 bp upstream of the original insertion site. R5 elements were identified at both sites. These findings expand the distribution of non-LTR elements that are specialized for insertion into the 28S gene and suggest that still more elements exist in other eukaryotic taxa. Attempts to trace the phylogeny of R5 did not offer sufficient resolution to determine whether R2, R4, and R5 represent the same lineage or whether they represent independent specializations for the 28S gene. PMID- 12777503 TI - SIRE1, an endogenous retrovirus family from Glycine max, is highly homogeneous and evolutionarily young. AB - SIRE1 is unusual among Ty1-copia retrotransposons in that it has an additional open reading frame with structural features similar to retroviral envelope proteins between pol and the 3' LTR. Here we report the characterization and comparison of eight different SIRE1 elements derived from a soybean genomic library, as well as SIRE1 reverse transcriptases from Glycine soja. The DNA sequences of the eight SIRE1 elements are highly homogeneous and share greater than 95% nucleotide identity. Partial sequences obtained from BAC ends are similarly conserved. Phylogenetic analyses resolve two closely related SIRE1 lineages, and nucleotide changes within and between SIRE1 lineages have occurred to preserve function. Both the gag and the env-like genes are evolving under similar levels of functional constraint. Considerable sequence heterogeneity in the form of short duplications was found within the LTRs and in the region between the envelope-like ORF and the 3' LTR. These duplications are suggestive of slippage by reverse transcriptase during replication. Sequence identity between LTRs of individual insertions suggests that they transposed within the past 70,000 years. Two of 10 SIRE1 insertions examined abut Ty3-gypsy retroelements. Since the soybean genome harbors more than 1,000 SIRE1 insertions, the collective data suggest that SIRE1 has undergone a very recent and robust amplification in soybean. PMID- 12777505 TI - Adaptation in the env gene of HIV-1 and evolutionary theories of disease progression. AB - The exact mechanisms by which HIV overwhelms the immune system remain poorly understood. Among the several explanations of HIV disease progression, most include adaptation of the viral genome to the host environment as a causal factor. Therefore, quantifying the rate and pattern of adaptive evolution within infected patients is critical to understanding the development of AIDS. Using sequence data from infected individuals sampled at multiple time points, I estimate the within-host adaptation rate of the HIV-1 env gene for viral populations from 50 different patients. I find that, averaging across patients, one adaptive substitution occurs every 3.3 months. Also, one adaptive mutation is driven to a high frequency (>50% but <100%) every 2.5 months. Taken together, such adaptive events occur once every 25 viral generations, which is the fastest adaptation rate ever recorded for a single protein-coding gene. Within the entire env gene, I estimate that a majority ( approximately 55%) of both nonsynonymous substitutions and high-frequency polymorphisms are adaptive. Further, in the C2 V5 region of env, I find that patients with longer asymptomatic periods have virus populations with higher adaptation rates, corroborating the notion that a broad, strong immune response against epitopes in the env gene product leads to longer asymptomatic periods. I conclude by discussing the distribution of nonsynonymous changes over the env gene. PMID- 12777504 TI - Parallel functional changes in the digestive RNases of ruminants and colobines by divergent amino acid substitutions. AB - A morphological or physiological trait may appear multiple times in evolution. At the molecular level, similar protein functions may emerge independently in different lineages. Whether these parallel functional changes are due to parallel amino acid substitutions has been a subject of debate. Here, I address this question using digestive ribonucleases (RNases) of two groups of foregut fermenting mammals: ruminant artiodactyls and colobine monkeys. The RNase1 gene was duplicated twice in ancestral ruminants at least 40 MYA, and it was also duplicated in the douc langur, an Asian colobine, approximately 4 MYA. After duplication, similar functional changes occurred in the ruminant and monkey enzymes. Interestingly, five amino acid substitutions in ruminant RNases that are known to affect its catalytic activity against double-stranded (ds) RNA did not occur in the monkey enzyme. Rather, a similar functional change in the monkey was caused by a different set of nine substitutions. Site-directed mutagenesis was used to make three of the five ruminant-specific substitutions in the monkey enzyme. Functional assays of these mutants showed that one of the three substitutions has a similar effect in monkeys, the second has a stronger effect, and the third has an opposite effect. These results suggest that (1) an evolutionary problem can have multiple solutions, (2) the same amino acid substitution may have opposite functional effects in homologous proteins, (3) the stochastic processes of mutation and drift play an important role even at functionally important sites, and (4) protein sequences may diverge even when their functions converge. PMID- 12777506 TI - The effect of positive selection on a sexual reproduction gene in Thalassiosira weissflogii (Bacillariophyta): results obtained from maximum-likelihood and parsimony-based methods. AB - Maximum-Likelihood-based and parsimony-based methods were used to test for potential effects of positive selection on the sexually induced gene 1 (Sig1) in Thalassiosira weissflogii. The Sig proteins are thought to play a role in mediating sperm-egg recognition during the sexual reproduction phase. The results obtained from parsimony-based analyses showed that none of the amino acid sites were influenced by positive selection. Maximum-likelihood analyses indicated that positive selection was affecting a maximum of seven and a minimum of four amino acid sites in the polypeptide derived from Sig1. It was concluded that the results obtained from the maximum-likelihood-based method are more reliable than those obtained from the parsimony-based approach. This is apparently the first study that has shown that reproductive proteins in unicellular eukaryotes are influenced by positive selection. PMID- 12777507 TI - Following the LINEs: an analysis of primate genomic variation at human-specific LINE-1 insertion sites. AB - The L1 Ta subfamily of long interspersed elements (LINEs) consists exclusively of human-specific L1 elements. Polymerase chain reaction-based screening in nonhuman primate genomes of the orthologous sites for 249 human L1 Ta elements resulted in the recovery of various types of sequence variants for approximately 12% of these loci. Sequence analysis was employed to capture the nature of the observed variation and to determine the levels of gene conversion and insertion site homoplasy associated with LINE elements. Half of the orthologous loci differed from the predicted sizes due to localized sequence variants that occurred as a result of common mutational processes in ancestral sequences, often including regions containing simple sequence repeats. Additional sequence variation included genomic deletions that occurred upon L1 insertion, as well as successive mobile element insertions that accumulated within a single locus over evolutionary time. Parallel independent mobile element insertions at orthologous loci in distinct species may introduce homoplasy into retroelement-based phylogenetic and population genetic data. We estimate the overall frequency of parallel independent insertion events at L1 insertion sites in seven different primate species to be very low (0.52%). In addition, no cases of insertion site homoplasy involved the integration of a second L1 element at any of the loci, but rather largely involved secondary insertions of Alu elements. No independent mobile element insertion events were found at orthologous loci in the human and chimpanzee genomes. Therefore, L1 insertion polymorphisms appear to be essentially homoplasy free characters well suited for the study of population genetics and phylogenetic relationships within closely related species. PMID- 12777508 TI - Estimating the distribution of selection coefficients from phylogenetic data with applications to mitochondrial and viral DNA. AB - The distribution of selection coefficients of new mutations is of key interest in population genetics. In this paper we explore how codon-based likelihood models can be used to estimate the distribution of selection coefficients of new amino acid replacement mutations from phylogenetic data. To obtain such estimates we assume that all mutations at the same site have the same selection coefficient. We first estimate the distribution of selection coefficients from two large viral data sets under the assumption that the viral population size is the same along all lineages of the phylogeny and that the selection coefficients vary among sites. We then implement several new models in which the lineages of the phylogeny may have different population sizes. We apply the new models to a data set consisting of the coding regions from eight primate mitochondrial genomes. The results suggest that there might be little power to determine the exact shape of the distribution of selection coefficient but that the normal and gamma distributions fit the data significantly better than the exponential distribution. PMID- 12777509 TI - Nonneutral admixture of immigrant genotypes in African Drosophila melanogaster populations from Zimbabwe. AB - Drosophila melanogaster originated in Africa and colonized the rest of the world only recently (approximately 10,000 to 15,000 years ago). Using 151 microsatellite loci, we investigated patterns of gene flow between African D. melanogaster populations representing presumptive ancestral variation and recently colonized European populations. Although we detected almost no evidence for alleles of non-African ancestry in a rural D. melanogaster population from Zimbabwe, an urban population from Zimbabwe showed evidence for admixture. Interestingly, the degree of admixture differed among chromosomes. X chromosomes of both rural and urban populations showed almost no non-African ancestry, but the third chromosome in the urban population showed up to 70% of non-African alleles. When chromosomes were broken into contingent microsatellite blocks, even higher estimates of admixture and significant heterogeneity in admixture was observed among these blocks. The discrepancy between the X chromosome and the third chromosome is not consistent with a neutral admixture hypothesis. The higher number of European alleles on the third chromosome could be due to stronger selection against foreign alleles on the X chromosome or to more introgression of (beneficial) alleles on the third chromosome. PMID- 12777510 TI - A codon-based model of host-specific selection in parasites, with an application to the influenza A virus. AB - Parasites sometimes expand their host range by acquiring a new host species. After a host change event, the selective regime acting on a given parasite gene may change as a result of host-specific adaptive alterations of protein functionality or host-specific immune-mediated selection. We present a codon based model that attempts to include these effects by allowing the position specific substitution process to change in conjunction with a host change event. Following maximum-likelihood parameter estimation, we employ an empirical Bayesian procedure to identify candidate sites potentially involved in host specific adaptation. We discuss the applicability of the model to the more general problem of ascertaining whether the selective regime differs in two groups of related organisms. The utility of the model is illustrated on a data set of nucleoprotein sequences from the influenza A virus obtained from avian and human hosts. PMID- 12777511 TI - Recently integrated Alu elements and human genomic diversity. AB - A comprehensive analysis of two Alu Y lineage subfamilies was undertaken to assess Alu-associated genomic diversity and identify new Alu insertion polymorphisms for the study of human population genetics. Recently integrated Alu elements (283) from the Yg6 and Yi6 subfamilies were analyzed by polymerase chain reaction (PCR), and 25 of the loci analyzed were polymorphic for insertion presence/absence within the genomes of a diverse array of human populations. These newly identified Alu insertion polymorphisms will be useful tools for the study of human genomic diversity. Our screening of the Alu insertion loci also resulted in the recovery of several "young" Alu elements that resided at orthologous positions in nonhuman primate genomes. Sequence analysis demonstrated these "young" Alu insertions were the products of gene conversion events of older, preexisting Alu elements or independent parallel forward insertions of older Alu elements in the same short genomic region. The level of gene conversion between Alu elements suggests that it may have an influence on the single nucleotide polymorphism within Alu elements in the genome. We have also identified two genomic deletions associated with the retroposition and insertion of Alu Y lineage elements into the human genome. This type of Alu retroposition mediated genomic deletion is a novel source of lineage-specific evolution within primate genomes. PMID- 12777512 TI - Origin and speciation of haplochromine fishes in East African crater lakes investigated by the analysis of their mtDNA, Mhc genes, and SINEs. AB - The Western Branch of the East African Great Rift Valley is pocketed with craters of extinct or dormant volcanoes. Many of the craters are filled with water, and the lakes are inhabited by fishes. The objective of the present study was to determine the amount and nature of genetic variation in haplochromine fishes inhabiting two of these crater lakes, Lake Lutoto and Lake Nshere, and to use this information to infer the origin and history of the two populations. To this end, sequences of mitochondrial (mt) DNA control region, exon 2 of major histocompatibility complex (Mhc) class II B genes, and short interspersed elements (SINEs) were analyzed. The results indicate that the Lake Nshere and Lake Lutoto fishes originated from different but related large founding populations derived from the Kazinga Channel, which connects Lake Edward and Lake George. Some of the genetic polymorphism that existed in the ancestral populations was lost in the populations of the two lakes. The polymorphism that has been retained has persisted for some 50000 generations (years). During this time, new mutations arose and became fixed in each of the two populations in the mtDNA, giving rise to sets of diagnostic substitutions. Each population evolved in isolation after the colonization of the lakes less than 50000 years ago. There appears to be no population structure within the crater lake fishes, and their present effective population sizes are in the order of 104 to 105 individuals. Comparisons with the endemic haplochromine species of Lake Victoria reveal interesting parallels, as well as differences, which may help to understand the nature of the speciation process. PMID- 12777513 TI - Antiquity and evolution of the MADS-box gene family controlling flower development in plants. AB - MADS-box genes in plants control various aspects of development and reproductive processes including flower formation. To obtain some insight into the roles of these genes in morphological evolution, we investigated the origin and diversification of floral MADS-box genes by conducting molecular evolutionary genetics analyses. Our results suggest that the most recent common ancestor of today's floral MADS-box genes evolved roughly 650 MYA, much earlier than the Cambrian explosion. They also suggest that the functional classes T (SVP), B (and Bs), C, F (AGL20 or TM3), A, and G (AGL6) of floral MADS-box genes diverged sequentially in this order from the class E gene lineage. The divergence between the class G and E genes apparently occurred around the time of the angiosperm/gymnosperm split. Furthermore, the ancestors of three classes of genes (class T genes, class B/Bs genes, and the common ancestor of the other classes of genes) might have existed at the time of the Cambrian explosion. We also conducted a phylogenetic analysis of MADS-domain sequences from various species of plants and animals and presented a hypothetical scenario of the evolution of MADS-box genes in plants and animals, taking into account paleontological information. Our study supports the idea that there are two main evolutionary lineages (type I and type II) of MADS-box genes in plants and animals. PMID- 12777514 TI - Horizontal gene transfer accelerates genome innovation and evolution. AB - Horizontal gene transfer (HGT) spreads genetic diversity by moving genes across species boundaries. By rapidly introducing newly evolved genes into existing genomes, HGT circumvents the slow step of ab initio gene creation and accelerates genome innovation. However, HGT can only affect organisms that readily exchange genes (exchange communities). In order to define exchange communities and understand the internal and external environmental factors that regulate HGT, we analyzed approximately 20,000 genes contained in eight free-living prokaryotic genomes. These analyses indicate that HGT occurs among organisms that share similar factors. The most significant are genome size, genome G/C composition, carbon utilization, and oxygen tolerance. PMID- 12777515 TI - Hop, an active Mutator-like element in the genome of the fungus Fusarium oxysporum. AB - A new type of active DNA transposon has been identified in the genome of Fusarium oxysporum by its transposition into the niaD target gene. Two insertions within the final exon, in opposite orientations at the same nucleotide site, have been characterized. These elements, called Hop, are 3,299 bp long, with perfect terminal inverted repeats (TIRs) of 99 bp. The sequencing of genomic copies reveals a 9-bp target site duplication and no apparent sequence specificity at the insertion sites. The sequencing of a cDNA indicates that Hop does not contain an intron and encodes a putative transposase of 836 amino acids. The structural features (length, TIRs size, and 9-bp duplication), together with the presence of conserved domains in the transposase, strongly suggest that Hop is a Mutator-like element (MULE). Hop is thus the first active member of this family found beyond plants. The high rate of excision observed indicates that Hop is very active and thus represents a promising efficient tagging system for the isolation of fungal genes. The distribution of Hop elements within the Fusarium genus revealed that they are present in different species, suggesting that related elements could be present in other fungal genomes. In fact, Hop-related sequences have been identified in the survey of the entire genome sequence of three other ascomycetes, Magnaporthe grisea, Neurospora crassa, and Aspergillus fumigatus. PMID- 12777516 TI - More taxa, more characters: the hoatzin problem is still unresolved. AB - The apparently rapid and ancient diversification of many avian orders complicates the resolution of their relationships using molecular data. Recent studies based on complete mitochondrial DNA (mtDNA) sequences or shorter lengths of nuclear sequence have helped corroborate the basic structure of the avian tree (e.g., a basal split between Paleognathae and Neognathae) but have made relatively little progress in resolving relationships among the many orders within Neoaves. We explored the potential of a moderately sized mtDNA data set ( approximately 5000 bp for each of 41 taxa), supplemented with data from a nuclear intron ( approximately 700 bp per taxon), to resolve relationships among avian orders. Our sampling of taxa addresses two issues: (1). the sister relationship and monophyly, respectively, of Anseriformes and Galliformes and (2). relationships of the enigmatic hoatzin Opisthocomus hoazin. Our analyses support a basal split between Galloanserae and Neoaves within Neognathae and monophyly of both Galliformes and Anseriformes. Within Galliformes, megapodes and then cracids branch basally. Within Anseriformes, mitochondrial data support a screamer (Anhimidae) plus magpie goose (Anseranatidae) clade. This result, however, may be an artifact of divergent base composition in one of the two anatids we sampled. With deletion of the latter taxon, Anseranas is sister to anatids as in traditional arrangements and recent morphological studies. Although our data provide limited resolution of relationships within Neoaves, we find no support for a sister relationship between either cuckoos (Cuculiformes) or turacos (Musophagiformes) and hoatzin. Both mitochondrial and nuclear data are consistent with a relationship between hoatzin and doves (Columbiformes), although this result is weakly supported. We also show that mtDNA sequences reported in another recent study included pervasive errors that biased the analysis towards finding a sister relationship between hoatzin and turacos. PMID- 12777517 TI - Using a pericentromeric interspersed repeat to recapitulate the phylogeny and expansion of human centromeric segmental duplications. AB - Despite considerable advances in sequencing of the human genome over the past few years, the organization and evolution of human pericentromeric regions have been difficult to resolve. This is due, in part, to the presence of large, complex blocks of duplicated genomic sequence at the boundary between centromeric satellite and unique euchromatic DNA. Here, we report the identification and characterization of an approximately 49-kb repeat sequence that exists in more than 40 copies within the human genome. This repeat is specific to highly duplicated pericentromeric regions with multiple copies distributed in an interspersed fashion among a subset of human chromosomes. Using this interspersed repeat (termed PIR4) as a marker of pericentromeric DNA, we recovered and sequence-tagged 3 Mb of pericentromeric DNA from a variety of human chromosomes as well as nonhuman primate genomes. A global evolutionary reconstruction of the dispersal of PIR4 sequence and analysis of flanking sequence supports a model in which pericentromeric duplications initiated before the separation of the great ape species (>12 MYA). Further, analyses of this duplication and associated flanking duplications narrow the major burst of pericentromeric duplication activity to a time just before the divergence of the African great ape and human species (5 to 7 MYA). These recent duplication exchange events substantially restructured the pericentromeric regions of hominoid chromosomes and created an architecture where large blocks of sequence are shared among nonhomologous chromosomes. This report provides the first global view of the series of historical events that have reshaped human pericentromeric regions over recent evolutionary time. PMID- 12777518 TI - Loss of DNA recombinational repair enzymes in the initial stages of genome degeneration. AB - Many obligate intracellular pathogens and symbionts undergo genome degeneration during long-term association with eukaryotic hosts; however, very little is known about genome changes that occur in the initial stages of such intracellular associations. By focusing on a clade of bacteria that have recently established symbiotic associations with insect hosts, we have identified events that may contribute to the reduction and degeneration of symbiont genomes. Unlike virtually all other bacteria, the obligate symbionts of maize and rice weevils each display substantial sequence divergence between multiple copies of their rDNA genes, resulting from a reduction in the efficacy of recombinational gene conversion, coincident with the inactivation of the recombinational repair gene recF in the common ancestor of both symbionts. The maize weevil endosymbiont also lacks a functional recA, resulting in further reduction in the efficacy of gene conversion between paralogous rDNAs and in a novel IS-mediated deletion in a 23S rDNA gene. Similar events may be pervasive during the evolution of symbiosis because symbiont genomes typically lack recombinational repair genes and have reduced numbers of ribosomal operons. PMID- 12777519 TI - Obtaining maximal concatenated phylogenetic data sets from large sequence databases. AB - To improve the accuracy of tree reconstruction, phylogeneticists are extracting increasingly large multigene data sets from sequence databases. Determining whether a database contains at least k genes sampled from at least m species is an NP-complete problem. However, the skewed distribution of sequences in these databases permits all such data sets to be obtained in reasonable computing times even for large numbers of sequences. We developed an exact algorithm for obtaining the largest multigene data sets from a collection of sequences. The algorithm was then tested on a set of 100,000 protein sequences of green plants and used to identify the largest multigene ortholog data sets having at least 3 genes and 6 species. The distribution of sizes of these data sets forms a hollow curve, and the largest are surprisingly small, ranging from 62 genes by 6 species, to 3 genes by 65 species, with more symmetrical data sets of around 15 taxa by 15 genes. These upper bounds to sequence concatenation have important implications for building the tree of life from large sequence databases. PMID- 12777520 TI - Genome-wide patterns of expression in Drosophila pure species and hybrid males. AB - One of the most fundamental questions for understanding the origin of species is why genes that function to cause fertility in a pure-species genetic background fail to produce fertility in a hybrid genetic background. A related question is why the sex that is most often sterile or inviable in hybrids is the heterogametic (usually male) sex. In this survey, we have examined the extent and nature of differences in gene expression between fertile adult males of two Drosophila species and sterile hybrid males produced from crosses between these species. Using oligonucleotide microarrays and real-time quantitative polymerase chain reaction, we have identified and confirmed that differences in gene expression exist between pure species and hybrid males, and many of these differences are quantitative rather than qualitative. Furthermore, genes that are expressed primarily or exclusively in males, including several involved in spermatogenesis, are disproportionately misexpressed in hybrids, suggesting a possible genetic cause for their sterility. PMID- 12777521 TI - Phylogenetic network and physicochemical properties of nonsynonymous mutations in the protein-coding genes of human mitochondrial DNA. AB - Theories on molecular evolution predict that phylogenetically recent nonsynonymous mutations should contain more non-neutral amino acid replacements than ancient mutations. We analyzed 840 complete coding-region human mitochondrial DNA (mtDNA) sequences for nonsynonymous mutations and evaluated the mutations in terms of the physicochemical properties of the amino acids involved. We identified 465 distinct missense and 6 nonsense mutations. 48% of the amino acid replacements changed polarity, 26% size, 8% charge, 32% aliphaticity, 13% aromaticity, and 44% hydropathy. The reduced-median networks of the amino acid changes revealed relatively few differences between the major continent-specific haplogroups, but a high variation and highly starlike phylogenies within the haplogroups. Some 56% of the mutations were private, and 25% were homoplasic. Nonconservative changes were more common than expected among the private mutations but less common among the homoplasic mutations. The asymptotic maximum of the number of nonsynonymous mutations in European mtDNA was estimated to be 1,081. The results suggested that amino acid replacements in the periphery of phylogenetic networks are more deleterious than those in the central parts, indicating that purifying selection prevents the fixation of some alleles. PMID- 12777522 TI - Pseudogenes contribute to the extreme diversity of nuclear ribosomal DNA in the hard coral Acropora. AB - One characteristic of Indo-Pacific Acropora spp. is extremely high diversity in rDNA sequences at both the species and individual levels. In order to test the hypothesis that pseudogenes may contribute to this diversity, three kinds of analyses were conducted. First, for A. millepora (the species containing the most diverse suite of rDNA types), RT-PCR was used to determine which 5.8S rDNA types are expressed. Second, as previous studies have indicated that interspecific hybridization has occurred in the genus Acropora and silencing of rDNA loci via nucleolar dominance has been shown in some cases to involve methylation, patterns of variation were examined at methylation-susceptible sites. Third, patterns of substitution at conserved sites (including those that are likely to contribute to secondary structure in rRNA) in the 5.8S rDNA were examined. These analyses consistently indicated that one rDNA sequence type present in a broad range of Indo-Pacific Acropora species is likely to consist predominantly of pseudogenes. Patterns of variation also suggest that species may differ with respect to which rDNA sequence types have been silenced and which are active. These pseudogenes are likely to have arisen as a consequence of the introduction of highly divergent rDNA types into single genomes by interspecific hybridization events, and we attribute the extreme rDNA diversity characteristic of many Acropora species to both the independent evolution of these silenced rDNA types and to the suppressive effects of high sequence diversity on homogenization processes acting on functional loci. PMID- 12777523 TI - Adaptive evolution in the photosensory domain of phytochrome A in early angiosperms. AB - Flowering plant diversity now far exceeds the combined diversity of all other plant groups. Recently identified extant remnants of the earliest-diverging lines suggest that the first angiosperms may have lived in shady, disturbed, and moist understory habitats, and that the aquatic habit also arose early. This would have required the capacity to begin life in dimly lit environments. If so, evolution in light-sensing mechanisms may have been crucial to their success. The photoreceptor phytochrome A is unique among angiosperm phytochromes in its capacity to serve a transient role under conditions where an extremely high sensitivity is required. We present evidence of altered functional constraints between phytochrome A (PHYA) and its paralog, PHYC. Tests for selection suggest that an elevation in nonsynonymous rates resulted from an episode of selection along the branch leading to all angiosperm PHYA sequences. Most nucleotide sites (95%) are selectively constrained, and the ratio of nonsynonymous to synonymous substitutions on branches within the PHYA clade does not differ from the ratio on the branches in the PHYC clade. Thus, positive selection at a handful of sites, rather than relaxation of selective constraints, apparently has played a major role in the evolution of the photosensory domain of phytochrome A. The episode of selection occurred very early in the history of flowering plants, suggesting that innovation in phyA may have given the first angiosperms some adaptive advantage. PMID- 12777524 TI - Molecular phylogenies and divergence times of sea urchin species of Strongylocentrotidae, Echinoida. AB - Sea urchins of the family Strongylocentrotidae have been important model systems in many fields of basic biology, yet knowledge of their evolutionary identities such as the phylogenetic relationships and divergence times remains limited. Here, I inferred molecular phylogenies of seven Strongylocentrotid species (Strongylocentrotus franciscanus, S. nudus, S. purpuratus, S. intermedius, S. droebachiensis, S. pallidus, and Hemicentrotus pulcherrimus) from the analyses of mitochondrial DNA sequences of 12SrDNA (349 nt), 12SrDNA-tRNA(gln) region (862 nt), and a combined sequence of cytochrome oxidase subunit I (COI, 1080 nt) and NADH dehydrogenase subunit I (NDI, 742 nt). The rate of sequence evolution and divergence times for each species were then estimated from the trees with reference to the time of separation between Strongylocentrotidae and Parechinidae, 35 to 50 MYA. The three trees agree well with each other, and the phylogeny is summarized by ((S. franciscanus, S. nudus), (H. pulcherrimus (S. purpuratus, S. intermedius (S. droebachiensis, S. pallidus)))). It is notable that the genus Strongylocentrotus consists of two distinct clades and that H. pulcherrimus branches off within Strongylocentrotus, implying assignment of a separate, monospecific genus to this species inappropriate. The rate of sequence evolution is calibrated to be 0.24%-0.34%/Myr in 12SrDNA, 0.25%-0.36%/Myr in 12SrDNA-tRNA(gln), and 0.65%-0.93%/Myr in COI-NDI combined sequences. S. purpuratus, in particular, shows the significantly higher rate of evolution in the 12SrDNA and 12SrDNA-tRNA(gln) regions compared to other species, suggesting careful use of its sequences in comparative studies. The two clades of Strongylocentrotidae seem to have split 13-19 MYA, and H. pulcherrimus branched off 7.2-14 MYA. In the former clade, S. franciscanus and S. nudus separated 5.7 8.1 MYA. In the latter clade, S. purpuratus, S. intermedius, and the clade of S. droebachiensis and S. pallidus diverged approximately 4.6-12 MYA, and the last two closest species separated 2.1-3.1 MYA. PMID- 12777525 TI - The major tick salivary gland proteins and toxins from the soft tick, Ornithodoros savignyi, are part of the tick Lipocalin family: implications for the origins of tick toxicoses. AB - The origins of tick toxicoses remain a subject of controversy because no molecular data are yet available to study the evolution of tick-derived toxins. In this study we describe the molecular structure of toxins from the soft tick, Ornithodoros savignyi. The tick salivary gland proteins (TSGPs) are four highly abundant proteins proposed to play a role in salivary gland granule biogenesis of the soft tick O. savignyi, of which the toxins TSGP2 and TSGP4 are a part. They were assigned to the lipocalin family based on sequence similarity to known tick lipocalins. Several other tick lipocalins were also identified using Smith Waterman database searches, bringing the tick lipocalin family up to 20. Phylogenetic analysis showed that most tick lipocalins group within genus specific clades, suggesting that gene duplication and divergence of tick lipocalin function occurred after tick speciation, most probably during the evolution of a hematophagous lifestyle. TSGP2 and TSGP3 show high sequence identity and group terminal to moubatin, an inhibitor of collagen-induced platelet aggregation from the tick, O. moubata. However, no platelet aggregation inhibitory activity is associated with the TSGPs using ADP or collagen as agonists, suggesting that TSGP2 and TSGP3 duplicated after divergence of O. savignyi and O. moubata. This timing is supported by the absence of TSGP2-4 in the salivary gland extracts of O. moubata. The absence of TSGP2 and TSGP4 in salivary gland extracts from O. moubata correlates with the nontoxicity of this tick species. The implications of this study are that the various forms of tick toxicoses do not have a common origin, but must have evolved independently in those tick species that cause pathogenesis. PMID- 12777526 TI - Systematic phylogenomic evidence of en bloc duplication of the ancestral 8p11.21 8p21.3-like region. AB - The genomes of many higher organisms, including plants and bony fish, frequently undergo polyploidization, and it has long been hypothesized that these, and other, large-scale genomic duplications have played an important role in the major evolutionary transitions of our past. Here we build upon an early work to show that the human genomic region 8p11.21-8p21.3 has three paralogous regions on chromosomes 4, 5, and 10 that were produced by two rounds of duplications after the protostomian-deuterostomian split and before the actinopterygian sarcopterygian split. We base our analysis on the phylogenetic reconstruction of the evolutionary history of 38 gene families located in these regions. Using an alignment centered on protein domains, three different phylogenetic methods, and divergence time estimation, this analysis gives more support in favor of two ancient polyploidization events in the vertebrate ancestral genome. PMID- 12777527 TI - Predicting demographic group structures based on DNA sequence data. AB - The ability to infer relationships between groups of sequences, either by searching for their evolutionary history or by comparing their sequence similarity, can be a crucial step in hypothesis testing. Interpreting relationships of human immunodeficiency virus type 1 (HIV-1) sequences can be challenging because of their rapidly evolving genomes, but it may also lead to a better understanding of the underlying biology. Several studies have focused on the evolution of HIV-1, but there is little information to link sequence similarities and evolutionary histories of HIV-1 to the epidemiological information of the infected individual. Our goal was to correlate patterns of HIV 1 genetic diversity with epidemiological information, including risk and demographic factors. These correlations were then used to predict epidemiological information through analyzing short stretches of HIV-1 sequence. Using standard phylogenetic and phenetic techniques on 100 HIV-1 subtype B sequences, we were able to show some correlation between the viral sequences and the geographic area of infection and the risk of men who engage in sex with men. To help identify more subtle relationships between the viral sequences, the method of multidimensional scaling (MDS) was performed. That method identified statistically significant correlations between the viral sequences and the risk factors of men who engage in sex with men and individuals who engage in sex with injection drug users or use injection drugs themselves. Using tree construction, MDS, and newly developed likelihood assignment methods on the original 100 samples we sequenced, and also on a set of blinded samples, we were able to predict demographic/risk group membership at a rate statistically better than by chance alone. Such methods may make it possible to identify viral variants belonging to specific demographic groups by examining only a small portion of the HIV-1 genome. Such predictions of demographic epidemiology based on sequence information may become valuable in assigning different treatment regimens to infected individuals. PMID- 12777528 TI - The molecular evolution of catalatic hydroperoxidases: evidence for multiple lateral transfer of genes between prokaryota and from bacteria into eukaryota. AB - The past decade has produced an increasing number of reports on horizontal gene transfer between prokaryotic organisms. Only recently, with the flood of available whole genome sequence data and a renewed intensity of the debate about the universal tree of life, a very few reports on lateral gene transfer (LGT) from prokaryotes into the Eukaryota have been published. We have investigated and report here on the molecular evolution of the gene families that encode catalatic hydroperoxidases. We have found that this process included not only frequent horizontal gene transfer among prokaryotes but also several lateral gene transfer events between bacteria and fungi and between bacteria and the protistan ancestor of the alga/plant lineage. PMID- 12777529 TI - Molecular basis and evolutionary origins of color diversity in great star coral Montastraea cavernosa (Scleractinia: Faviida). AB - Natural pigments are normally products of complex biosynthesis pathways where many different enzymes are involved. Corals and related organisms of class Anthozoa represent the only known exception: in these organisms, each of the host tissue colors is essentially determined by a sequence of a single protein, homologous to the green fluorescent protein (GFP) from Aequorea victoria. This direct sequence-color linkage provides unique opportunity for color evolution studies. We previously reported the general phylogenetic analysis of GFP-like proteins, which suggested that the present-day diversity of reef colors originated relatively recently and independently within several lineages. The present work was done to get insight into the mechanisms that gave rise to this diversity. Three colonies of the great star coral Montastraea cavernosa (Scleractinia, Faviida) were studied, representing distinct color morphs. Unexpectedly, these specimens were found to express the same collection of GFP like proteins, produced by at least four, and possibly up to seven, different genetic loci. These genes code for three basic colors-cyan, green, and red-and are expressed differently relative to one another in different morphs. Phylogenetic analysis of the new sequences indicated that the three major gene lineages diverged before separation of some coral families. Our results suggest that color variation in M. cavernosa is not a true polymorphism, but rather a manifestation of phenotypic plasticity (polyphenism). The family level depth of its evolutionary roots indicates that the color diversity is adaptively significant. Relative roles of gene duplication, gene conversion, and point mutations in its evolution are discussed. PMID- 12777530 TI - Evidence that the human X chromosome is enriched for male-specific but not female specific genes. AB - There is increasing evidence that X chromosomes have an unusual complement of genes, especially genes that have sex-specific expression. However, whereas in worm and fly the X chromosome has a dearth of male-specific genes, in mice genes that are uniquely expressed in spermatogonia are especially abundant on the X chromosome. Is this latter enrichment true for nongermline, male-specific genes in mammals, and is it found also for female-specific genes? Here, using SAGE data, we show (1) that tissue-specific genes tend to be more abundant on the human X chromosome, (2) that, controlling for this effect, genes expressed exclusively in prostate are enriched on the human X chromosome, and (3) that genes expressed exclusively in mammary gland and ovary are not so enriched. This we propose is consistent with Rice's model of the evolution of sexually antagonistic alleles. PMID- 12777531 TI - Functional respiratory chain analyses in murid xenomitochondrial cybrids expose coevolutionary constraints of cytochrome b and nuclear subunits of complex III. AB - The large number of extant Muridae species provides the opportunity of investigating functional limits of nuclear/mitochondrial respiratory chain (RC) subunit interactions by introducing mitochondrial genomes from progressively more divergent species into Mus musculus domesticus mtDNA-less (rho0) cells. We created a panel of such xenomitochondrial cybrids, using as mitochondrial donors cells from six murid species with divergence from M. m. domesticus estimated at 2 to 12 Myr before present. Species used were Mus spretus, Mus caroli, Mus dunni, Mus pahari, Otomys irroratus, and Rattus norvegicus. Parsimony analysis of partial mtDNA sequences showed agreement with previous molecular phylogenies, with the exception that Otomys did not nest within the murinae as suggested by some recent nuclear gene analyses. Cellular production of lactate, a sensitive indicator of decreased respiratory chain ATP production, correlated with divergence. Functional characterization of the chimeric RC complexes in isolated mitochondria using enzymological analyses demonstrated varying decreases in activities of complexes I, III, and IV, which have subunits encoded in both mitochondrial and nuclear genomes. Complex III showed a striking decline in electron transfer function in the most divergent xenocybrids, being greatly reduced in the Rattus xenocybrid and virtually absent in the Otomys xenocybrid. This suggests that nuclear subunits interacting with cytochrome b face the greatest constraints in the coevolution of murid RC subunits. We sequenced the cytochrome b gene from the species used to identify potential amino acid substitutions involved in such interactions. The greater sensitivity of complex III to xenocybrid dysfunction may result from the encoding of redox center apoproteins in both nuclear and mitochondrial genomes, a unique feature of this RC complex. PMID- 12777532 TI - Three neuropeptide Y receptor genes in the spiny dogfish, Squalus acanthias, support en bloc duplications in early vertebrate evolution. AB - It has been debated whether the increase in gene number during early vertebrate evolution was due to multiple independent gene duplications or synchronous duplications of many genes. We describe here the cloning of three neuropeptide Y (NPY) receptor genes belonging to the Y1 subfamily in the spiny dogfish, Squalus acanthias, a cartilaginous fish. The three genes are orthologs of the mammalian subtypes Y1, Y4, and Y6, which are located in paralogous gene regions on different chromosomes in mammals. Thus, these genes arose by duplications of a chromosome region before the radiation of gnathostomes (jawed vertebrates). Estimates of duplication times from linearized trees together with evidence from other gene families supports two rounds of chromosome duplications or tetraploidizations early in vertebrate evolution. The anatomical distribution of mRNA was determined by reverse-transcriptase PCR and was found to differ from mammals, suggesting differential functional diversification of the new gene copies during the radiation of the vertebrate classes. PMID- 12777534 TI - Retroids in archaea: phylogeny and lateral origins. AB - Until recently, none of the diverse elements bearing reverse transcriptase (retroids) have been known from Archaea. However, in the recently published genomes of the acetate-utilizing archaeal methanogens, Methanosarcina acetivorans and M. mazei, several open reading frames (ORFs) are annotated as reverse transcriptase (RT). These annotations led us to the characterization of a retron and 13 retrointrons, including three twintrons, clustered at seven loci of the M. acetivorans genome, and four retrointrons at two loci of the M. mazei genome. Based on a phylogeny of the RT ORFs, we infer four lateral gene transfers (LGT) of these retroids from Bacteria to Archaea and of retrointron mobility within the Archaea genomes. Our phylogenetic analysis also identifies several novel retrons from GenBank in the bacterial groups Firmicutes, Fusobacteria, Cyanobacteria and beta-Proteobacteria, as well as in M. acetivorans. The discovery of retrointrons in Archaea as a consequence of LGT from Bacteria suggests that they did not originate in the progenote and parallels the "mitochondrial seed" theory of the origin of spliceosomes. Extending the known phylogenetic distribution of retroids to Archaea is consistent with the view that they have played a significant role in evolution of genomes throughout the tree of life. PMID- 12777535 TI - Assessing the phylogenetic utility of four mitochondrial genes and a nuclear intron in the asian pit viper genus, Trimeresurus: separate, simultaneous, and conditional data combination analyses. AB - A number of methods have been proposed for addressing how to optimize the analysis of multiple data sets from diverse mitochondrial and nuclear gene partitions in the pursuit of robust organismal phylogenies. The present study used separate, simultaneous, and conditional data combination methods to analyze 3,135 bp of data from four mitochondrial partitions and the seventh intron of the beta fibrinogen gene in the Asian pit viper genus, Trimeresurus sensu stricto. The phylogenetic utility and homogeneity of all partitions were estimated via a combination of homogeneity partition tests, homoplasy indices, and partitioned Bremer support. Despite the detection of significant heterogeneity of phylogenetic signal between the mitochondrial and nuclear partitions, the simultaneous analysis represented the best-supported topology of all the data. The relatively slow rate (approximately one quarter of the rate of mtDNA) and functionally unconstrained molecular evolution of the intron resulted in much lower levels of homoplasy compared with the mitochondrial partitions. This was further shown via partitioned Bremer support, which, when considered throughout hierarchical clade levels, highlighted the phylogenetic strength and limitations of the intron at deeper and shallower phylogenetic levels, respectively. The simultaneous analysis helped to resolve the phylogenetic relationships of taxa that were unresolved throughout all individual gene trees and tentatively supports the existence of morphologically and genetically distinct clades within the genus. Topological appraisals of the mitochondrial gene partitions suggest that the cytochrome b and the NADH subunit 4 gene partitions are better estimators of phylogenetic relationships than are the 12S and 16S ribosomal RNA partitions at the taxonomic levels under consideration. PMID- 12777536 TI - Rapid divergence of microsatellite abundance among species of Drosophila. AB - Among major taxonomic groups, microsatellites exhibit considerable variation in composition and allele length, but they also show considerable conservation within many major groups. This variation may be explained by slow microsatellite evolution so that all species within a group have similar patterns of variation, or by taxon-specific mutational or selective constraints. Unfortunately, comparing microsatellites across species and studies can be problematic because of biases that may exist among different isolation and analysis protocols. We present microsatellite data from five Drosophila species in the Drosophila subgenus: D. arizonae, D. mojavensis, and D. pachea (three cactophilic species), and D. neotestacea and D. recens (two mycophagous species), all isolated at the same time using identical protocols. For each species, we compared the relative abundance of motifs, the distribution of repeat size, and the average number of repeats. Dimers were the most abundant microsatellites for each species. However, we found considerable variation in the relative abundance of motif size classes among species, even between sister taxa. Frequency differences among motifs within size classes for the three cactophilic species, but not the two mycophagous species, are consistent with other studied Drosophila. Frequency distributions of repeat number, as well as mean size, show significant differences among motif size classes but not across species. Sizes of microsatellites in these five species are consistent with D. virilis, another species in the subgenus Drosophila, but they have consistently higher means than in D. melanogaster, in the subgenus Sophophora. These results confirm that many aspects of microsatellite variation evolve quickly but also are subject to taxon specific constraints. In addition, the nature of microsatellite evolution is dependent on temporal and taxonomic scales, and some variation is conserved across broad taxonomic levels despite relatively high rates of mutation for these loci. PMID- 12777537 TI - Current concepts in minimal access surgery for children. AB - Since the introduction of minimal access surgery to general surgeons in the 1980s, pediatric surgeons have been employing this innovative technology to perform surgery on children. Video technology and miniaturized instruments have brought the laboratory to the operating room; in many cases several small incisions are the only access necessary to perform complicated procedures that would otherwise require a large wound. Additional benefits of minimal access surgery may include reduced postoperative analgesic requirements, shortened length of stay, and faster resumption of normal activities. Increased operative costs offset some of these gains. The pediatric surgical community has embraced minimal access techniques for some operations; others remain controversial. PMID- 12777538 TI - International Small for Gestational Age Advisory Board consensus development conference statement: management of short children born small for gestational age, April 24-October 1, 2001. AB - OBJECTIVE: To provide pediatric endocrinologists, general pediatricians, neonatologists, and primary care physicians with recommendations for the management of short children born small for gestational age (SGA). METHODS: A 13 member independent panel of pediatric endocrinologists was convened to discuss relevant issues with respect to definition, diagnosis, and clinical management of short children born SGA. Panel members convened over a series of 3 meetings to thoroughly review, discuss, and come to consensus on the identification and treatment of short children who are born SGA. CONCLUSIONS: SGA is defined as birth weight and/or length at least 2 standard deviations (SDs) below the mean for gestational age (2 SD below the mean; this catch up process is usually completed by the time they are 2 years of age. A child who is SGA and older than 3 years and has persistent short stature (ie, remaining at least 2 SD below the mean for chronologic age) is not likely to catch up and should be referred to a pediatrician who has expertise in endocrinology. Bone age is not a reliable predictor of height potential in children who are SGA. Nevertheless, a standard evaluation for short stature should be performed. A diagnosis of SGA does not exclude growth hormone (GH) deficiency, and GH assessment should be performed if there is clinical suspicion or biochemical evidence of GH deficiency. At baseline, insulin-like growth factor-I, insulin like growth factor binding protein-3, fasting insulin, glucose, and lipid levels as well as blood pressure should be measured, and all aspects of SGA-not just stature-should be addressed with parents. The objectives of GH therapy in short children who are SGA are catch-up growth in early childhood, maintenance of normal growth in childhood, and achievement of normal adult height. GH therapy is effective and safe in short children who are born SGA and should be considered in those older than 2 to 3 years. There is long-term experience of improved growth using a dosage range from 0.24 to 0.48 mg/kg/wk. Higher GH doses (0.48 mg/kg/wk [0.2 IU/kg/d]) are more effective for the short term. Whether the higher GH dose is more efficacious than the lower dose in terms of adult height results is not yet known. Only adult height results of randomized dose-response studies will give a definite answer. Monitoring is necessary to ensure safety of medication. Children should be monitored for changes in glucose homeostasis, lipids, and blood pressure during therapy. The frequency and intensity of monitoring will vary depending on risk factors such as family history, obesity, and puberty. PMID- 12777539 TI - Isolated hepatocyte transplantation in an infant with a severe urea cycle disorder. AB - OBJECTIVE: Transplantation of isolated hepatocytes in animal models has been shown to correct inborn errors of metabolism. Based on these studies and our experience with hepatocyte transplantation in a child with Crigler-Najjar syndrome, isolated hepatocyte transplantation was performed to attempt metabolic reconstitution in a male infant with severe ornithine transcarbamylase (OTC) deficiency. METHODS: An infant with an antenatal diagnosis of OTC deficiency was managed intensively to prevent hyperammonemia. Isolated hepatocytes were obtained by collagenase perfusion of donated livers not used for transplantation. Hepatocytes were infused in batches over the first 4 weeks of life via an umbilical venous catheter positioned in the portal vein. Immunosuppression consisted of tacrolimus and corticosteroids. RESULTS: Over 4 billion viable hepatocytes were transplanted during the first 3.5 weeks of life. A period of metabolic stability was achieved between days 20 and 31 during which normal protein intake was tolerated while phenylbutyrate was weaned. During this time, plasma ammonia and glutamine remained within normal limits. Hyperammonemia reappeared abruptly on day 31 of life. Protein tolerance diminished to baseline; metabolic stability was subsequently reattained only following successful liver transplantation at 6 months of age. CONCLUSIONS: Isolated hepatocyte transplantation appeared to result in temporary relief of hyperammonemia and protein intolerance attributable to OTC deficiency. The metabolic stability achieved was lost after 11 days presumably because of rejection of the transplanted cells because of insufficient immunosuppression. Future attempts at isolated hepatocyte transplantation for inborn errors of metabolism in humans should include adequate immunosuppression and a liver biopsy as a means of proving hepatocyte engraftment and function. PMID- 12777540 TI - Adult patient visits to children's hospital emergency departments. AB - BACKGROUND: Adults frequently seek medical services in children's hospital emergency departments (CHEDs), and are required to be admitted to CHEDs under the provisions of the Emergency Medical Transfer and Active Labor Act (EMTALA), which requires medical evaluation and stabilization of every patient who presents to an emergency department. In recent years visits by adults to CHEDs appear to have increased. OBJECTIVE: There were 3 objectives to the current study: 1) to examine secular trends in the number of adult patients visiting CHEDs, 2) to determine if perceived increases are related to the implementation of EMTALA, and 3) to examine the characteristics, evaluation, and disposition of adult patients presenting for first-time visits to a CHED. METHODS: A database of all visits to an urban CHED between 1992 and 2002 was queried to collect information on adult patients (22 years or older). New adult patients were identified based on the assignment of new medical record numbers. The medical records of all adult patients presenting during the 1-year interval before and after the institution's full implementation of EMTALA were reviewed and relevant data collected. RESULTS: Over the study period, there were 501,033 patient visits to the CHED. Of these, 5512 (1.1%) were by adult patients, which included 536 (9.7%) new adult patients. Using the chi(2), test we found a significant increase in the total number of adult visits and the number of new adult visits, particularly after the implementation of EMTALA. The mean age of the new adult patients was 34.9 +/- 11.9 years. Their most frequent chief complaints were injuries (24.4%), cardiac related problems (7.6%), and syncope (6.7%). A total of 427 (79.7%) of the new adult patients were treated and released, 81 (15.1%) were transferred to an outside hospital for additional care, and 15 (2.8%) were admitted to our hospital. There were no significant differences between the new adult populations in 1997 and 1999. Comparing new and established adult populations in 1999, the population of new adults was significantly older (28.1 +/- 6.8 vs 34.9 +/- 11.9 years) and more likely to present with injuries or syncopal episodes. Among the total cohort of new adult patients in the study, chest pain also occurred at a significantly higher rate compared with established adults (6.7% vs 3.8%). CONCLUSIONS: Adult visits to CHEDs appear to be increasing in frequency in association with the implementation of EMTALA regulations. It is therefore essential that physicians staffing CHEDs be properly trained in the stabilization of common adult medical emergencies. We recommend that the language of EMTALA be revised to allow adult patients with nonemergent problems to be directly referred to adult emergency departments, which are more appropriate than CHEDs. PMID- 12777533 TI - Gene diversity patterns at 10 X-chromosomal loci in humans and chimpanzees. AB - We have investigated the pattern and extent of nucleotide diversity in 10 X chromosomal genes where mutations are known to cause mental retardation in humans. For each gene, we sequenced the entire coding region from cDNA in humans, chimpanzees, and orangutans, as well as about 3 kb of genomic DNA in 20 humans sampled worldwide and in 10 chimpanzees representing two "subspecies." Overall nucleotide diversity in these genes is about twofold lower in humans than in chimpanzees, and nucleotide diversity within and between species is low, suggesting that a high level of functional constraint acts on these genes. Strikingly, we find that a summary of the allele frequency spectrum is significantly correlated in humans and chimpanzees, perhaps reflecting very similar levels of constraint at these genes in the two species. A possible exception is FMR2, which shows a higher number of nonsynonymous than synonymous substitutions on the human lineage, suggesting the action of positive selection. PMID- 12777541 TI - The risks of adverse neonatal outcome among preterm small for gestational age infants according to neonatal versus fetal growth standards. AB - OBJECTIVE: To evaluate neonatal and fetal growth standards in determining the impact of small for gestational age (SGA) on neonatal mortality and morbidity. DESIGN: A hospital-based cohort study of infants born in a regional tertiary care center and admitted to the neonatal intensive care unit. SETTING AND PARTICIPANTS: A total of 1267 singleton neonates of <34 weeks gestational age, without any congenital anomalies, born between January 1, 1993 and December 31, 2001. OUTCOME MEASURES: Each outcome variable including mortality, respiratory distress syndrome, bronchopulmonary dysplasia, intraventricular hemorrhage (IVH), periventricular leukomalacia, and necrotizing enterocolitis was related to growth status as defined by fetal and neonatal growth standards after adjustment for potential confounders. RESULTS: The number of SGA infants was 11.6% (n = 147) of the study population according to neonatal growth standards, but it was increased to 23.3% (n = 295) when fetal growth standards were used. According to fetal growth standards, when SGA was compared with appropriate for gestational age infants, it was associated with an increased risk of respiratory distress syndrome (odds ratio [OR] 1.40; 95% confidence interval [CI] 1.00-1.95), bronchopulmonary dysplasia (OR 2.18; 95% CI 1.33-3.59), IVH (OR 1.67; 95% CI 1.13 2.45), and retinopathy of prematurity (OR 3.88; 95% CI 2.33-6.48). However, only neonatal mortality (OR 3.64; 95% CI 1.64-8.09), retinopathy of prematurity (OR 5.38; 95% CI 2.87-10.90), and necrotizing enterocolitis (OR 2.47; 95% CI 1.21 5.07) were positively associated with SGA when using neonatal growth standards. CONCLUSIONS: Compared with the neonatal growth standards, the fetal growth standards are better in identifying increased risk of respiratory morbidity and IVH among preterm SGA infants. PMID- 12777542 TI - Successful promotion of hepatitis B vaccinations among Vietnamese-American children ages 3 to 18: results of a controlled trial. AB - OBJECTIVE: Chronic infection with the hepatitis B virus is endemic in Southeast Asian populations, including Vietnamese. Previous research has documented low rates of hepatitis B vaccine coverage among Vietnamese-American children and adolescents ages 3 to 18. To address this problem, we designed and tested in a controlled trial 2 public health outreach "catch-up" campaigns for this population. DESIGN: In the Houston, Texas metropolitan area, we mounted a media led information and education campaign, and in the Dallas metropolitan area, we organized a community mobilization strategy. We evaluated the success of these interventions in a controlled trial, using the Washington, DC metropolitan area as a control site. To do so, we conducted computer-assisted telephone interviews with random samples of approximately 500 Vietnamese-American households in each of the 3 study sites both before and after the interventions. We assessed respondents' awareness and knowledge of hepatitis B and asked for hepatitis B vaccination dates for a randomly selected child in each household. When possible, we validated vaccination dates through direct contact with each child's providers. RESULTS: Awareness of hepatitis B increased significantly between the pre- and postintervention surveys in all 3 areas, and the increase in the media education area (+21.5 percentage points) was significantly larger than in the control area (+9.0 percentage points). At postintervention, significantly more parents knew that free vaccines were available for children in the media education (+31.9 percentage points) and community mobilization (+16.7 percentage points) areas than in the control area (+4.7 percentage points). An increase in knowledge of sexual transmission of hepatitis B virus was significant in the media education area (+14.0 percentage points) and community mobilization (+13.6 percentage points) areas compared with the control area (+5.2 percentage points). Parent- or provider-reported data (n = 783 for pre- and n = 784 for postintervention surveys) suggest that receipt of 3 hepatitis B vaccinations increased significantly in the community mobilization area (from 26.6% at pre- to 38.8% at postintervention) and in the media intervention area (28.5% at pre- and 39.4% at postintervention), but declined slightly in the control community (37.8% at pre- and 33.5% at postintervention). Multiple logistic regression analyses estimated that the odds of receiving 3 hepatitis B vaccine doses were significantly greater for both community mobilization (odds ratio 2.15, 95% confidence interval 1.16-3.97) and media campaign (odds ratio 3.02, 95% confidence interval 1.62-5.64) interventions compared with the control area. The odds of being vaccinated were significantly greater for children who had had at least 1 diphtheria-tetanus-pertussis shot, and whose parents were married, knew someone with liver disease, had heard of hepatitis B, and had greater knowledge about hepatitis B. The odds of being vaccinated were significantly lower for older children. CONCLUSIONS: Both community mobilization and media campaigns significantly increased the knowledge of Vietnamese-American parents about hepatitis B vaccination, and the receipt of "catch-up" vaccinations among their children. PMID- 12777543 TI - Economic analysis of promotion of hepatitis B vaccinations among Vietnamese American children and adolescents in Houston and Dallas. AB - OBJECTIVE: To ascertain the cost-effectiveness and benefit-cost ratios of 2 public health campaigns conducted in Dallas and Houston in 1998-2000 for "catch up" hepatitis B vaccination of Vietnamese-Americans born 1984-1993. DESIGN: Program evaluation. SETTING: Houston and Dallas, Texas. PARTICIPANTS: A total of 14,349 Vietnamese-American children and adolescents. INTERVENTIONS: Media-led information and education campaign in Houston, and community mobilization strategy in Dallas. Outcomes were compared with a control site: Washington, DC. MAIN OUTCOME MEASURES: Receipt of 1, 2, or 3 doses of hepatitis B vaccine before and after the interventions, costs of interventions, cost-effectiveness ratios for intermediate outcomes, intervention cost per discounted year of life saved, and benefit-cost ratio of the interventions. RESULTS: The number of children who completed the series of 3 hepatitis B vaccine doses increased by 1176 at a total cost of 313,904 dollars for media intervention, and by 390 and at 169,561 dollars for community mobilization. Costs per child receiving any dose, per dose, and per completed series were 363 dollars, 101 dollars, and 267 dollars for media intervention and 387 dollars, 136 dollars, and 434 dollars for community mobilization, respectively. For media intervention, the intervention cost per discounted year of life saved was 9954 dollars and 131 years of life were saved; for community mobilization, estimates were 11,759 dollars and 60 years of life. The benefit-cost ratio was 5.26:1 for media intervention and 4.47:1 for community mobilization. CONCLUSION: Although the increases in the number of children who completed series of 3 doses were modest for both the Houston and Dallas areas, both media education and, to a lesser degree, community mobilization interventions proved cost-effective and cost-beneficial. PMID- 12777544 TI - Counting the shots: a model for immunization screening and referral in nonmedical settings. AB - BACKGROUND: Clinics of the Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) have become important partners in efforts to improve vaccination coverage in low income children. However, the time required to assess all antigens in each child's vaccination record may exceed the capacity of many of these clinics. Seeking a solution, experts recommended assessing up-to-date (UTD) status only for the diphtheria-tetanus-acellular-pertussis (DTaP) vaccine and treating this as a proxy measure for all vaccines in the childhood schedule. Whether this single vaccine screening method represents an acceptable alternative to the traditional multiple-vaccine method as a basis for improving overall immunization coverage levels in this vulnerable population has not been demonstrated. OBJECTIVE: To evaluate the validity of the proposed simplified method for assessing immunization status in a nationally representative population of infants and children who had ever been enrolled in WIC before 35 months old. METHODS: This was a cross-sectional analysis of the 2000 National Immunization Survey representing children ages 3 to 24 months who had ever been enrolled in WIC. For the 6277 children in the study population, we compared personal records of completion status for DTaP with personal records of completion status for all immunizations appropriate for age in the combination 4:3:1:3 schedule to see which of the 2 (single vs multiple screening) methods would better predict the child's true (provider-reported) status for the 4:3:1:3 series. The main outcome measures were the comparative sensitivity, specificity, and overall test efficiency of the 2 methods in correctly identifying underimmunized WIC children. RESULTS: Completion status for DTaP was less sensitive than completion status for all vaccinations in correctly identifying truly underimmunized children (sensitivity = 70% and 77%, respectively). However, it was more specific in correctly identifying children who were truly UTD for age (specificity = 86% and 82%, respectively). The 2 methods were essentially identical with respect to overall test efficiency (82% and 81% for DTaP assessment and assessment of all vaccines, respectively). CONCLUSIONS: Given limited resources to do immunization screening and referral in nonmedical settings such as WIC, simplifying the process by using DTaP from the personal vaccination record as a proxy for the 4:3:1:3 series is a viable option. Loss in sensitivity may well be offset by gains in the capacity of WIC clinics to screen more children. PMID- 12777545 TI - Infants with bilirubin levels of 30 mg/dL or more in a large managed care organization. AB - OBJECTIVE: To describe the incidence, etiology, treatment, and outcome of newborns with total serum bilirubin (TSB) levels >or=30 mg/dL (513 micro mol/L). DESIGN: Population-based case series. SETTING: Eleven Northern California Kaiser Permanente Medical Care Program hospitals and 1 affiliated hospital. PATIENTS: Eleven infants with TSB levels of >or=30 mg/dL in the first 30 days after birth, identified using computer databases from a cohort of 111,009 infants born 1995 1998. OUTCOME MEASURES: Clinical data from the birth hospitalization, rehospitalization, and outpatient visits in all infants; psychometric testing at age 5 (N = 3), neurologic examinations by child neurologists at age 5 (N = 3), or primary care providers (N = 7; mean age: 2.2 years); Parent Evaluation of Developmental Status (N = 8; mean age: 4.2 years). RESULTS: Maximum TSB levels of the 11 infants ranged from 30.7 to 45.5 mg/dL (525 micro mol/L to 778 micro mol/L; mean: 34.9 mg/dL [597 micro mol/L]). Four were born at 35 to 36 weeks gestation, and 7 were exclusively breastfed. Two had apparent isoimmunization; the etiology for the other 9 remained obscure, although only 4 were tested for glucose-6-phosphate dehydrogenase deficiency and 1 was bacteremic. None had acute neurologic symptoms. All received phototherapy and 5 received exchange transfusions. One infant died of sudden infant death syndrome; there was no kernicterus at autopsy. Two were lost to follow-up but were neurologically normal when last seen for checkups at 18 and 43 months. One child was receiving speech therapy at age 3. There were no significant parental concerns or abnormalities in the other children. CONCLUSIONS: In this setting, TSB levels >or=30 mg/dL were rare and generally unaccompanied by acute symptoms. Although we did not observe serious neurodevelopmental sequelae in this small sample, additional studies are required to quantify the known, significant risk of kernicterus in infants with very high TSB levels. PMID- 12777546 TI - Psychosocial health among young victims and offenders of direct and indirect bullying. AB - OBJECTIVE: To assess the association between bullying (both directly and indirectly) and indicators of psychosocial health for boys and girls separately. STUDY DESIGN: A school-based questionnaire survey of bullying, depression, suicidal ideation, and delinquent behavior. SETTING: Primary schools in Amsterdam, The Netherlands. PARTICIPANTS: A total of 4811 children aged 9 to 13. RESULTS: Depression and suicidal ideation are common outcomes of being bullied in both boys and girls. These associations are stronger for indirect than direct bullying. After correction, direct bullying had a significant effect on depression and suicidal ideation in girls, but not in boys. Boy and girl offenders of bullying far more often reported delinquent behavior. Bullying others directly is a much greater risk factor for delinquent behavior than bullying others indirectly. This was true for both boys and girls. Boy and girl offenders of bullying also more often reported depressive symptoms and suicidal ideation. However, after correction for both sexes only a significant association still existed between bullying others directly and suicidal ideation. CONCLUSIONS: The association between bullying and psychosocial health differs notably between girls and boys as well as between direct and indirect forms of bullying. Interventions to stop bullying must pay attention to these differences to enhance effectiveness. PMID- 12777547 TI - Smoking during pregnancy and newborn neurobehavior. AB - OBJECTIVE: This was a prospective study of the effects of maternal smoking during pregnancy on newborn neurobehavior, including dose-response relationships using self-report and a bioassay of nicotine exposure. METHODS: The sample included 27 nicotine exposed and 29 unexposed full-term newborn infants with no medical problems from comparable social class backgrounds. Mothers were excluded for using illegal drugs during pregnancy, using antidepressant medication, or if they consumed >3 alcoholic drinks per month. Nicotine exposure was determined by maternal self-report and cotinine in maternal saliva. The NICU Network Neurobehavioral Scale (NNNS) was administered by masked examiners in hospital to measure neurobehavioral function. NNNS scores were compared between nicotine exposed and -unexposed groups including adjustment for covariates. Dose-response relationships with NNNS scores were computed for maternal salivary cotinine and maternal report of number of cigarettes per day during pregnancy. RESULTS: After adjustment for covariates, the tobacco-exposed infants were more excitable and hypertonic, required more handling and showed more stress/abstinence signs, specifically in the central nervous system (CNS), gastrointestinal, and visual areas. Dose-response relationships showed higher maternal salivary cotinine values related to more stress/abstinence signs (r =.530) including CNS (r =.532) and visual stress (r =.688) and higher excitability scores (r =.617). Cigarettes per day during pregnancy was related to more stress/abstinence signs (r =.582) including CNS (r =.561) and visual stress (r =.640). CONCLUSIONS: These findings suggest neurotoxic effects of prenatal tobacco exposure on newborn neurobehavior. Dose-response relationships could indicate neonatal withdrawal from nicotine. Research directed at understanding the effects of cigarette smoking during pregnancy on infants can lead to improved public health outcome. PMID- 12777548 TI - The effect of a parenting education program on the use of preventive pediatric health care services among low-income, minority mothers: a randomized, controlled study. AB - OBJECTIVE: To determine if a community-based intervention program focusing on parenting education will have an impact on preventive health care utilization behaviors among low-income, minority mothers in Washington, DC. DESIGN: The experimental design was a randomized, controlled study in which 286 mother-infant dyads were assigned to either the standard social services (control) group or to the intervention group. Women and their newborn infants were recruited during the immediate postpartum period in 4 Washington, DC, hospital sites from April 1995 to April 1997. The year-long multicomponent intervention included home visits and hospital-based group sessions in addition to the standard social services available at the hospital sites. A total of 286 postpartum women with inadequate prenatal care were assigned randomly to the control or the intervention group. Women and their infants were followed for 1 full year. Outcome measures included usage of preventive health care services including well care infant visits and adherence to immunization schedules during the first year of the infant's life. RESULTS: Infants in the intervention group initiated well care at an earlier age than controls (by 6 weeks, 62.5% vs 50% had received their first well infant visit). Infants in the intervention group had more frequent well visits (by 12 months of age, 3.5 vs 2.7 visits). Multivariate analyses showed infants in the intervention group to be more likely to complete their scheduled immunizations (by 9 months, odds ratio = 2.2, 95% confidence interval: 1.09-4.53). Those in the intervention group with more frequent contacts (30+ visits) with study personnel were most likely to have followed age-appropriate immunization schedules when compared with controls (at 9 months odds ratio = 3.63, 95% confidence interval: 1.58-8.33). CONCLUSIONS: It is possible to influence health care usage patterns of high-risk minority populations through public health interventions that are global in their perspective. Focusing on parental knowledge and beliefs regarding health-related issues and life skills in a self-efficacy model is associated with improved usage of infant health care resources. PMID- 12777549 TI - Evaluation of cytomegalovirus infections transmitted via breast milk in preterm infants with a real-time polymerase chain reaction assay. AB - OBJECTIVE: Preterm infants are at greater risk of symptomatic cytomegalovirus (CMV) infection than term infants. Breast milk is the main source of perinatal CMV infections. This study evaluated the kinetics of CMV load in breast milk and the rate of postnatal CMV transmission via breast milk from mothers to their preterm infants. METHODS: This was a prospective study of 30 mothers and their 43 preterm infants. The infants either had a gestational age of <34 weeks or weighed <2000 g at birth. Breast milk, serum, and urine samples were collected every 2 weeks until discharge, and screened for CMV infection using a real-time PCR assay. Most of the breast milk had been preserved at -20 degrees C before feeding to the preterm infants. RESULTS: Twenty-four mothers (24 of 30, 80%), who had 34 preterm infants, were CMV immunoglobulin G positive. Twenty-one (87.5%) of the 24 seropositive mothers, who had 30 preterm infants, had detectable CMV deoxyribonucleic acid (DNA) in breast milk during the study period. Most breast milk became positive for CMV DNA 2 weeks after delivery. Viral DNA copy numbers increased until they peaked at 4 to 6 weeks. Afterward, the CMV DNA copy numbers decreased. Of the 30 infants who were fed CMV DNA-positive breast milk, CMV infection was confirmed in 3 infants. However, they had no clinical symptoms of CMV infection. CONCLUSIONS: Despite the high rate of CMV DNA in breast milk, symptomatic infections in the preterm infants did not occur. These results might be associated with the method of breast milk preservation and the population we studied. CMV infections transmitted via breast milk feeding did not have much impact on preterm infants in our institutes. PMID- 12777550 TI - Initiation of breastfeeding among mothers of very low birth weight infants. AB - OBJECTIVE: To examine factors that predict the initiation of expressed milk feedings and the transition to direct breastfeedings among mothers of very low birth weight (VLBW) infants. METHODS: The sample consists of 361 mother-infant pairs enrolled in a follow-up study of children aged 6 to 8 years who were born weighing <1501 g in 1 of 5 hospitals between 1991-1993. Chart review at birth provided data on neonatal characteristics and demographic factors at delivery were obtained by postpartum maternal interview. Information regarding infant feeding practices was obtained at follow-up. RESULTS: In this study, 60% of mothers initiated expressed milk feedings for their VLBW infants. However, the duration of these feedings was brief with 52% of infants receiving 1 to 3 months or less of human milk feedings. Greater educational attainment, private insurance, and breastfeeding experience were each independently associated with the decision to provide expressed milk feedings. Only 27% of mothers reported directly breastfeeding their VLBW infants. The transition from expressed milk feedings to direct breastfeedings was positively associated with sociodemographic factors including maternal age, insurance status, and breastfeeding experience as well as the length of hospitalization, an indicator of infant health. CONCLUSIONS: Sociodemographic factors were associated with both the decision to initiate expressed milk feedings and the transition to direct breastfeedings. However, factors relating to infant health only influenced the transition to direct breastfeedings. Intervention programs need to consider the sociodemographic factors that influence infant feeding decisions as well as specific challenges encountered by mothers of VLBW infants. PMID- 12777551 TI - Appendectomy during childhood and adolescence and the subsequent risk of cancer in Sweden. AB - OBJECTIVE: Researchers have speculated that surgical excision of lymphoid tissue, such as appendectomy, early in life might confer an increased risk of cancer. In this study, we determined the risks of cancer for people who had appendectomy performed during childhood. METHODS: We studied the risk of cancer in a large Swedish cohort of children who had appendectomy performed during the period of 1965-1993. Standardized incidence ratios (SIRs) were computed using age-, gender , and period-specific incidence rates derived from the entire Swedish population as comparison. Hospital discharge diagnosis data were used to examine cancer risks by categories of surgery, medical conditions, and type of appendicitis. The average length of follow-up was 11.2 years. RESULTS: We found no excess overall cancer risk but noted a significant excess for stomach cancer (SIR: 2.45; 95% confidence interval [CI]: 1.1-4.8) and a borderline increase of non-Hodgkin's lymphoma (NHL; SIR: 1.55; 95% CI: 1.0-2.3). The elevated risks for both cancers were only evident 15 or more years after appendectomy (stomach cancer, SIR: 3.82; 95% CI: 1.7-7.5; NHL, SIR: 2.49; 95% CI: 1.4-4.2). CONCLUSIONS: It is reassuring that there was no overall increase of cancer several years after childhood appendectomy. Increased risks for NHL and stomach cancer, occurring 15 or more years after appendectomy, were based on small absolute numbers of excess cancers. As 95% of the subjects were younger than 40 years at exit, this cohort requires continuing follow-up and monitoring. PMID- 12777552 TI - Pediatricians' reported practices regarding early education and Head Start referral. AB - BACKGROUND: Early learning programs have proven benefits for impoverished children; Head Start is the most widespread of such programs. The current involvement of pediatricians in the Head Start enrollment process is unknown. OBJECTIVES: 1) To assess the knowledge, attitudes, and reported practices of pediatricians on referring families to Head Start; 2) to assess pediatricians' receptivity to a potential practice-based intervention to enhance their ability to make Head Start referrals. METHODS: Mail survey to stratified random sample of pediatricians practicing in poor and non-poor US zip codes. Prevalence estimates and logistic regression models were estimated using weighted data. RESULTS: Of 1000 surveys distributed, 472 of 772 presumed-eligible subjects completed surveys for a response rate of 61%. Respondents and nonrespondents were similar with regard to age, gender, years in practice, and urban/rural practice setting. Eighty percent of pediatricians reported discussing child care arrangements with a majority of their preschool-aged patients' families, while only 14% reported actually assisting these families in applying to Head Start. Lack of time (77% of pediatricians) and nonphysician office staff (71%) were listed as the most significant barriers to helping families apply to Head Start. Unfamiliarity with early childhood education (10%) was generally not seen as a barrier to this practice. Head Start knowledge (adjusted odds ratio [aOR]: 1.43; 95% confidence interval [CI]: 1.01, 2.02), self-efficacy in advising families how to access local Head Start programs (aOR: 3.49; 95% CI: 1.46, 8.38), and the belief that it is the pediatrician's responsibility to do so (aOR: 9.98; 95% CI: 3.91, 25.48) were significantly associated with assisting families with Head Start enrollment. The majority of respondents (77%) reported a willingness to participate in a proposed computer-based intervention to aid eligible families in applying to Head Start. Having access to a social worker (aOR: 2.48; 95% CI: 1.17, 5.21) and respondent age (aOR: 0.96 for each year; 95% CI: 0.93, 0.99) were significantly associated with likely participation in the intervention. CONCLUSIONS: Although pediatricians report commonly discussing child care issues, few actively assist patients in the application process for Head Start. An intervention to facilitate Head Start referral from the physician's office must address time and staff limitations; education of pediatricians is a secondary need. PMID- 12777553 TI - Patient safety events during pediatric hospitalizations. AB - OBJECTIVE: Our objective was to describe potential patient safety events for hospitalized children, using the patient safety indicators (PSIs), and examine associations with these events. METHODS: PSI algorithms, developed by researchers at the Agency for Healthcare Research and Quality to identify potential in hospital patient safety problems using administrative data, were applied to 3.8 million discharge records for children under 19 years from 22 states in the 1997 Healthcare Cost and Utilization Project. Prevalence of PSI events and associations with patient-level and hospital-level characteristics, length of stay, in-hospital mortality, and total charges were examined. RESULTS: The prevalence of pediatric patient safety events is significant with the highest rate found for birth trauma at 1.5 cases per every 100 births. The majority of these events for birth trauma consist of long bone and skull fractures, excluding the clavicle. Compared with records without PSI events, discharges with PSI events had 2- to 6-fold longer lengths of stay, 2- to 18-fold higher rates of in hospital mortality, and 2- to 20-fold higher total charges. Bivariate and multivariate analyses found that all PSI events except birth trauma were directly associated with factors related to greater severity of illness and large urban teaching institutions. Birth trauma, however, was directly associated with black and Hispanic ethnicity but was not consistently associated with technologically sophisticated teaching institutions. CONCLUSIONS: The prevalence of birth trauma and other potential patient safety events for hospitalized children is high and comparable to hospitalized adults. These events are associated with increased length of stay, in-hospital mortality, and total charges. Associated factors differ significantly for birth trauma compared with other PSI events. Institutional application of the PSIs may be useful to identify processes of care that warrant further evaluation as the health care industry tackles the problem of patient safety, particularly for children. PMID- 12777554 TI - Risk of sudden infant death syndrome and week of gestation of term birth. AB - OBJECTIVE: We sought to determine if the risk of sudden infant death syndrome (SIDS) varied according to week of delivery at term among elective and nonelective births. DESIGN: Retrospective cohort study. PARTICIPANTS: All single infants live born between 37 and 42 weeks gestation in Scotland between 1992 and 1995 documented in the Scottish Morbidity Record. OUTCOME: Death in the first year of life where SIDS was in the principal position on the death certificate. RESULTS: There were 202,622 eligible births and 119 deaths attributed to SIDS. Among infants delivered electively, there was no significant association between risk of SIDS and week of delivery. Among those delivered nonelectively, the risk of SIDS declined significantly with each week of gestation (odds ratio .72, 95% confidence interval .60-.86). This trend was only minimally attenuated by adjustment for maternal age, parity, smoking and socioeconomic deprivation category, infant sex, Apgar score, mode of delivery, and birth weight decile (adjusted odds ratio .78, 95% confidence interval .65-.93). CONCLUSIONS: We hypothesize that early spontaneous labor at term and SIDS may be linked because of a common association with suboptimal intrauterine environment. PMID- 12777555 TI - Health-related quality of life in urban elementary schoolchildren. AB - BACKGROUND: Health disparities between children from urban minority backgrounds and children from more affluent backgrounds are well-recognized. Few studies specifically address urban children's perceptions of their health-related quality of life (HRQOL) or the factors that contribute to these perceptions. Since schools are pivotal to children's intellectual, social, and emotional development, school connectedness may be a factor that contributes to their perception of HRQOL. OBJECTIVE: To examine children's perceptions of HRQOL in an elementary school-based population of urban children. METHODS: The study population consisted of 2nd, 3rd, and 5th graders from 6 urban kindergarten to 8th grade schools and their parents. Children completed a survey that included questions on HRQOL and school connectedness. Parents completed a telephone survey that assessed demographics, the child's health, health care usage, and parental health status. Data on school absences and mobility from the computerized school database were linked to survey data. Bivariate analyses were used to evaluate the association between child report of HRQOL and collected variables, including school connectedness. Multivariable linear regression was conducted to identify the factors best predicting HRQOL in these urban children. RESULTS: Of the 1150 eligible students, parent and child survey data were available for 525 (45.6%). Fifty-one percent of students were male and 89% were black. Ninety-four percent of parents were female, 29% were married, and 62% had family incomes below 20,000 dollars per year. The mean total score for HRQOL was 67.2, with a possible range of 0 to 100 (higher scores reflecting better HRQOL). In the multivariable analysis, child grade, the relationship of the " parent" to the child, employment, family income, type/presence of insurance, and school connectedness were significantly associated with the HRQOL total score. CONCLUSIONS: Young urban children self-report low HRQOL scores and do so as early as the 2nd grade. These low scores, which reflect children's own perceptions of impaired psychological and physical health, have potential implications for the success of urban children in their learning environments. The association between HRQOL and school connectedness might suggest that health and educational programs that improve a child's attachment to school could result in improved perceptions of health by urban children. PMID- 12777556 TI - Occult head injury in high-risk abused children. AB - OBJECTIVE: Head injury is the leading cause of death in abused children under 2 years of age. Evidence for establishing guidelines regarding screening for occult head injury in a neurologically asymptomatic child with other evidence of abuse is lacking. This is particularly important given that many children with acute inflicted head injury have evidence of old injury when they are diagnosed. The primary aim of this study was to estimate the prevalence of occult head injury in a high-risk sample of abused children with normal neurologic examinations. The secondary aim was to describe characteristics of this population. METHODS: Children under 2 years of age admitted to an urban children's hospital between January 1998 and December 2001 with injuries suspicious for child abuse were eligible for this study if they had a normal neurologic examination on admission. Subjects were selected if they met 1 of the following "high-risk" criteria: rib fractures, multiple fractures, facial injury, or age <6 months. Subjects were excluded if they had a history of neurologic dysfunction, seizures, respiratory arrest, or if their initial physical examination revealed scalp injury. RESULTS: Of the 65 patients who met these criteria, 51 (78.5%) had a head computed tomography or magnetic resonance imaging in addition to skeletal survey. Of these 51 patients, 19 (37.3%, 95% confidence interval 24.2-50.4%) had an occult head injury. Injuries included scalp swelling (74%), skull fracture (74%), and intracranial injury (53%). All except 3 of the head-injured patients had at least a skull fracture or intracranial injury. Skeletal survey alone missed 26% (5/19) of the cases. Head-injured children were younger than non-head-injured children (median age 2.5 vs 5.1 months); all but 1 head-injured child was <1 year of age. Among the head-injured children, 72% came from single parent households, 37% had mothers whose age was <21 years, and 26% had a history of prior child welfare involvement in their families. Ophthalmologic examination was performed in 14 of the 19 cases; no retinal hemorrhages were noted. CONCLUSIONS: Our results support a recommendation for universal screening in neurologically asymptomatic abused children with any of the high-risk criteria used in this study, particularly if that child is under 1 year of age. Ophthalmologic examination is a poor screening method for occult head injury, and one should proceed directly to computed tomography or magnetic resonance imaging. Given the high prevalence of occult head injury detected in this study, further study is warranted to estimate the prevalence of occult head injury in lower risk populations of abused children. PMID- 12777557 TI - Differential influence of family history of hypertension and premature myocardial infarction on systolic blood pressure and left ventricular mass trajectories in youth. AB - OBJECTIVE: To examine the influence of genetic susceptibility to essential hypertension (EH) and the genetic susceptibility to premature myocardial infarction (MI) on longitudinal development of systolic blood pressure (SBP) and left ventricular mass (LVM) in youth. METHODS: Individual SBP and LVM growth curves across age were created for a sample of 745 subjects (age range: 4.9-27.5 years) and a sample of 687 subjects (age range: 8.2-27.5 years), respectively. Each sample had an approximately equal proportion of African American and European American males and females, with annual assessments over a 10-year period. Family history (FH) of EH and FH of premature MI were used as measures of genetic susceptibility to EH and to premature MI, respectively. Positive FH (FH(+)) of EH and of premature MI were defined, respectively, as verified EH in 1 or both biological parents, and verified MI in any biological parent or grandparent before 55 years of age. RESULTS: Subjects with an FH(+) of EH had higher SBP levels and stronger increases in SBP over time than subjects with a negative FH (FH(-)) of EH. Subjects with an FH(+) of EH also showed higher LVM levels than subjects with an FH(-) of EH. In addition, the effect of an FH(+) of EH on LVM was stronger in females than males. The effects of FH of EH on SBP and LVM could not be explained by differences in socioeconomic status, but the effect on LVM was no longer significant after adjustment for BMI. FH of MI had no significant effects on SBP or LVM. CONCLUSIONS: Effects of genetic susceptibility to EH on SBP and LVM trajectories were observed in childhood, whereas no such effects were found for FH of MI. Genetic markers of EH may improve the understanding of individual differences in susceptibility to develop hypertension and LV hypertrophy. PMID- 12777558 TI - Retropharyngeal abscess in children: clinical presentation, utility of imaging, and current management. AB - OBJECTIVE: We sought to describe the clinical presentation of patients with retropharyngeal abscess (RPA), utility of imaging studies, and implications on management. METHODS: A retrospective chart review was performed at a tertiary care, pediatric hospital with cases identified by a discharge diagnosis of RPA; posttraumatic RPA cases were excluded. Patients without confirmatory radiographic findings, fluoroscopy, or computed tomography (CT) were excluded. RESULTS: Sixty four cases involving 64 patients were studied. The median age of the patients was 36 months; 48 (75%) of the 64 patients were younger than 5 years. The most common chief complaints were neck pain (38%), fever (17%), sore throat (17%), neck mass (16%), and respiratory distress or stridor (5%). In 29 children (45%), it was noted that there was limitation of neck extension, in 23 (36.5%) torticollis, and in 8 (12.5%) limitation of neck flexion. The physical examination revealed stridor with wheezing in only 1 patient (1.5%) and wheezing in 1 other (1.5%). Twenty-seven patients (42%) underwent surgery; 37 (58%) were treated with antibiotics only. Performance of a surgical procedure was significantly associated with CT scan findings. Ten (37%) of 27 patients with defined abscess on CT scan were treated with antibiotics alone. There were no treatment failures in either the antibiotic-only group or the antibiotics-plus-surgery group. CONCLUSIONS: Children with RPA present with limitation of neck movement, especially difficulty extending their neck to look up. They rarely present with respiratory distress or stridor. CT scan is useful to distinguish patients with RPA from those with retropharyngeal cellulitis. Most patients with retropharyngeal cellulitis and some with RPA can be treated successfully without surgery. PMID- 12777559 TI - Expanded newborn screening for inborn errors of metabolism by electrospray ionization-tandem mass spectrometry: results, outcome, and implications. AB - OBJECTIVE: The aims of this study were to determine the impact of expanded newborn screening using tandem mass spectrometry (MS/MS) on the overall detection rate of inborn errors of metabolism in Germany and to assess the outcome for the patients that were diagnosed. METHODS: During the period of study, 250,000 neonates in a German population were investigated for 23 inborn errors of metabolism by electrospray ionization-MS/MS. The overall value of the screening program was estimated by 1) complete ascertainment of all positive tests; 2) definite assignment of all diagnoses including reconfirmation at 12 months; and 3) clinical follow-up of all detected patients in an overall interval of 42 months. The mean observation period was 13.5 months per child. RESULTS: In 106 newborns, confirmed inborn errors of metabolism were found. The disorders were classified as 50 classic forms and 56 variants. A total of 825 tests (0.33%) were false-positives. Seventy of the 106 newborns with confirmed disorders were judged to require treatment. Six children developed symptoms despite treatment. Three children had died. Among 9 children who became symptomatic before report of the results of screening, in 6 the diagnosis had been made in advance of the screening report. In evaluation of the screening program, 61 of the 106 identified children (58% of true-positives, or 1 of 4100 healthy newborns) were judged to have benefited from screening and treatment, because the diagnosis had not been made before screening. None of these infants had died and none developed psychomotor retardation or metabolic crisis during the follow-up period. CONCLUSIONS: The screening by MS/MS for up to 23 additional disorders has approximately doubled the detection rate compared with that achieved by the conventional methods used in Germany. This strategy represents valuable preventive medicine by enabling diagnosis and treatment before the onset of symptoms. PMID- 12777560 TI - Human metapneumovirus infection in the United States: clinical manifestations associated with a newly emerging respiratory infection in children. AB - OBJECTIVE: Respiratory tract infections are a leading cause of morbidity and mortality worldwide. Recently, a newly identified human respiratory virus, human metapneumovirus (hMPV), was reported by investigators in the Netherlands. We sought to determine whether hMPV was circulating in our community and to determine the clinical features associated with hMPV infection. METHODS: Respiratory specimens from children who were younger than 5 years and had a negative result for respiratory syncytial virus, influenza A and B, parainfluenza viruses 1 to 3, and adenovirus by direct fluorescent antibody test were screened for hMPV by reverse transcriptase-polymerase chain reaction. Samples were collected from October 30, 2001, to February 28, 2002. RESULTS: Of the 296 patients screened, 19 (6.4%) had evidence of hMPV infection. hMPV was identified in patients with either upper or lower respiratory tract infection or both. Clinical manifestations included wheezing, hypoxia, and abnormal findings on chest radiographs (eg, focal infiltrates, peribronchial cuffing). Nosocomial infection occurred in at least 1 patient. CONCLUSIONS: hMPV is circulating in the United States and is associated with respiratory tract disease in patients with respiratory illnesses not caused by respiratory syncytial virus, influenza, parainfluenza viruses, and adenovirus. Additional studies are required to define the epidemiology and the extent of disease in the general population caused by hMPV. PMID- 12777561 TI - Platelet count and sepsis in very low birth weight neonates: is there an organism specific response? AB - OBJECTIVE: Thrombocytopenia is commonly observed in very low birth weight (VLBW) neonates with sepsis. Specific platelet responses to different infectious agents have not been extensively characterized. The objectives of this study were to examine platelet counts and platelet indices in preterm neonates with culture proven sepsis to determine if there are organism-specific platelet responses. STUDY DESIGN: We analyzed a cohort of all VLBW neonates (birthweight <1500 g) born over a 4-year period and admitted to a single level III neonatal intensive care unit (N = 943). Thrombocytopenia was defined as a platelet count <100,000/mm(3). Platelet count, nadir, duration of thrombocytopenia, and mean platelet volume (MPV) were examined during episodes of culture-proven sepsis. Analysis of variance, Kruskal-Wallis, Mann-Whitney U, and chi(2) tests were used to compare groups, and data are expressed as mean +/- standard deviation. RESULTS: Sepsis was diagnosed in 154 (16%) of 943 patients in the study population. Of the sepsis episodes, 54% were associated with thrombocytopenia and 61% with an elevation in MPV. Infections were grouped by organism type: Gram positive bacteria (117/154, 76%), Gram-negative bacteria (24/154, 16%), and fungi (13/154, 8%). When compared with patients with Gram-positive sepsis, those with Gram-negative or fungal sepsis had a significantly lower initial platelet count, a lower platelet nadir, a higher incidence of thrombocytopenia, and a greater duration of thrombocytopenia. The decrease in platelet count from baseline was also significantly less in the Gram-positive infections than in the fungal infections. Although there was an overall increase in MPV from baseline, there were no differences between groups. CONCLUSIONS: In our population of VLBW infants, sepsis is frequently associated with thrombocytopenia and an elevation in MPV. However, fungal and Gram-negative pathogens are associated with a lower platelet count and more prolonged thrombocytopenia compared with Gram-positive pathogens. We conclude that common pathogens causing sepsis have different effects on platelet kinetics. PMID- 12777562 TI - National Kidney Foundation's Kidney Disease Outcomes Quality Initiative clinical practice guidelines for chronic kidney disease in children and adolescents: evaluation, classification, and stratification. AB - OBJECTIVES: A series of new guidelines has been developed by the National Kidney Foundation's Kidney Disease Outcomes Quality Initiative to improve the detection and management of chronic kidney disease (CKD). In most instances of CKD, the earliest manifestations of the disorder may be identified by relatively simple tests. Unfortunately, CKD is often "underdiagnosed," in part because of the absence of a common definition of CKD and a classification of the stages in its progression. The Kidney Disease Outcomes Quality Initiative clinical practice guidelines for CKD evaluation, classification, and stratification provide a basis to remedy these deficits. The specific goals of the guidelines described in this review are to provide: 1) an overview of the clinical practice guidelines as they pertain to children and adolescents, 2) a simple classification of the stages of CKD, and 3) a practical approach to the laboratory assessment of kidney disease in children and adolescents. METHODS: The guidelines were developed as part of an evidence-based evaluation of CKD and its consequences in patients of all ages. The data that were used to generate the guidelines in this article were extracted from a structured analysis of articles that reported on children with CKD. RESULTS AND CONCLUSIONS: This review presents the definition and 5-stage classification system of CKD developed by the work group assigned to develop the guidelines, and summarizes the major recommendations regarding the early detection of CKD. Major emphasis is placed on the identification of children and adolescents with CKD by measuring the protein-to-creatinine ratio in spot urine specimens and by estimating the glomerular filtration rate from serum creatinine using prediction equations. PMID- 12777563 TI - Methadone and breastfeeding: new horizons. PMID- 12777564 TI - The assessment of newborn size. PMID- 12777565 TI - The US smallpox vaccination plan. PMID- 12777566 TI - Opening the window into brain development in children more widely with magnetic resonance imaging. PMID- 12777567 TI - School readiness: an idea whose time has arrived. PMID- 12777568 TI - Midaortic syndrome in the fetus and premature newborn: a new etiology of nonimmune hydrops fetalis and reversible fetal cardiomyopathy. AB - Nonimmune hydrops fetalis is the final common pathway of many conditions that ultimately result in fetal anasarca. Even after extensive evaluation, the etiology of a small percentage of cases of hydrops remains unknown. We present a case of midaortic syndrome, also known as abdominal coarctation syndrome, in a fetus with hydrops and a severe cardiomyopathy. The clinical manifestations of midaortic syndrome in this fetus and premature newborn, including malignant hypertension and reversible cardiomyopathy, are detailed. The fetal pathophysiology of midaortic syndrome remains speculative, but likely includes fetal hypertension as the cause of cardiac dysfunction. To our knowledge, this is the first report of midaortic syndrome as an etiology for nonimmune hydrops fetalis. PMID- 12777569 TI - Transmission of Salmonella enterica serotype typhimurium DT104 to infants through mother's breast milk. AB - This study documents the first reported transmission of Salmonella enterica serotype Typhimurium definitive type 104 (DT104) to premature fraternal twins via their mother's breast milk. When premature twin neonates developed severe enteritis in the neonatal intensive care unit (NICU), stool samples and the mother's breast milk were cultured for the presence of Salmonella. Antibacterial susceptibility patterns were determined. Semiquantitative organism abundance data were retrospectively gathered on 54 stored breast milk samples collected on 34 different days using a rapid, real-time polymerase chain reaction (PCR) methodology (LightCycler PCR). Fecal samples from other infants in the NICU at that time were also tested. Pulsed-field gel electrophoresis (PFGE) was used to assess the genetic composition of the isolated organisms. The twins' neonatal stools and mother's breast milk cultures revealed a resistance pattern (R-type) to ampicillin, chloramphenicol, streptomycin, sulfonamides, and tetracycline. LightCycler PCR analysis of sequential breast milk samples confirmed this to be the likely source of transmission. In the subsequent outbreak investigation, none of the NICU surveillance fecal samples proved positive for this organism. The genetic composition of organisms isolated from the maternal breast milk was indistinguishable from those isolated from neonatal specimens as determined by PFGE. Antibiotic susceptibility tests coupled with PFGE patterns suggested that these Salmonella isolates were DT104. Because the prevalence of DT104 infections is rising in the United States, neonatologists should be aware of breast milk as a potential mode of transmission. PMID- 12777570 TI - Fatal malignant hyperthermia-like syndrome with rhabdomyolysis complicating the presentation of diabetes mellitus in adolescent males. AB - OBJECTIVE: This report describes a new fatal syndrome observed in adolescent males at the initial presentation of diabetes mellitus. The features include hyperglycemic hyperosmolar coma complicated by a malignant hyperthermia-like picture with fever, rhabdomyolysis, and severe cardiovascular instability. DESIGN: Case series. SETTING: Pediatric intensive care units of 3 tertiary care facilities in the United States. PATIENTS: Six adolescent males, 5/6 obese with acanthosis nigricans, 4/6 black. RESULTS: Four of 6 patients died. Four of 6 patients did not have significant ketosis. Six of 6 patients had increased temperature after the administration of insulin. CONCLUSIONS: The underlying etiology of this syndrome remains unclear. Possibilities include an underlying metabolic disorder such as a fatty acid oxidation defect, an unrecognized infection, exposure to an unknown toxin, or a genetic predisposition to malignant hyperthermia. Evaluation for all these possibilities and empiric treatment with dantrolene should be considered for this type of patient until this syndrome is better characterized. PMID- 12777571 TI - Advanced practice in neonatal nursing. AB - The advanced practice neonatal nurse's participation in newborn care continues to be accepted and supported by the American Academy of Pediatrics. Recognized categories of advanced practice neonatal nurse are the neonatal clinical nurse specialist and the neonatal nurse practitioner. Training and credentialing requirements have been updated recently and are endorsed in this revised statement. PMID- 12777572 TI - Radiation disasters and children. AB - The special medical needs of children make it essential that pediatricians be prepared for radiation disasters, including 1) the detonation of a nuclear weapon; 2) a nuclear power plant event that unleashes a radioactive cloud; and 3) the dispersal of radionuclides by conventional explosive or the crash of a transport vehicle. Any of these events could occur unintentionally or as an act of terrorism. Nuclear facilities (eg, power plants, fuel processing centers, and food irradiation facilities) are often located in highly populated areas, and as they age, the risk of mechanical failure increases. The short- and long-term consequences of a radiation disaster are significantly greater in children for several reasons. First, children have a disproportionately higher minute ventilation, leading to greater internal exposure to radioactive gases. Children have a significantly greater risk of developing cancer even when they are exposed to radiation in utero. Finally, children and the parents of young children are more likely than are adults to develop enduring psychologic injury after a radiation disaster. The pediatrician has a critical role in planning for radiation disasters. For example, potassium iodide is of proven value for thyroid protection but must be given before or soon after exposure to radioiodines, requiring its placement in homes, schools, and child care centers. Pediatricians should work with public health authorities to ensure that children receive full consideration in local planning for a radiation disaster. PMID- 12777573 TI - Pediatric exposure and potential toxicity of phthalate plasticizers. AB - Phthalates are plasticizers that are added to polyvinyl chloride (PVC) products to impart flexibility and durability. They are produced in high volume and generate extensive though poorly defined human exposures and unique childhood exposures. Phthalates are animal carcinogens and can cause fetal death, malformations, and reproductive toxicity in laboratory animals. Toxicity profiles and potency vary by specific phthalate. The extent of these toxicities and their applicability to humans remains incompletely characterized and controversial. Two phthalates, diethylhexyl phthalate (DEHP) and diisononyl phthalate (DINP), have received considerable attention recently because of specific concerns about pediatric exposures. Like all phthalates, DEHP and DINP are ubiquitous contaminants in food, indoor air, soils, and sediments. DEHP is used in toys and medical devices. DINP is a major plasticizer used in children's toys. Scientific panels, advocacy groups, and industry groups have analyzed the literature on DEHP and DINP and have come to different conclusions about their safety. The controversy exists because risk to humans must be extrapolated from animal data that demonstrate differences in toxicity by species, route of exposure, and age at exposure and because of persistent uncertainties in human exposure data. This report addresses sensitive endpoints of reproductive and developmental toxicity and the unique aspects of pediatric exposures to phthalates that generate concern. DEHP and DINP are used as specific examples to illustrate the controversy. PMID- 12777574 TI - Postexposure prophylaxis in children and adolescents for nonoccupational exposure to human immunodeficiency virus. AB - Exposure to human immunodeficiency virus (HIV) can occur in a number of situations unique to, or more common among, children and adolescents. Guidelines for postexposure prophylaxis (PEP) for occupational and nonoccupational (eg, sexual, needle-sharing) exposures to HIV have been published by the US Public Health Service, but they do not directly address nonoccupational HIV exposures unique to children (such as accidental exposure to human milk from a woman infected with HIV or a puncture wound from a discarded needle on a playground), and they do not provide antiretroviral drug information relevant to PEP in children. This clinical report reviews issues of potential exposure of children and adolescents to HIV and gives recommendations for PEP in those situations. The risk of HIV transmission from nonoccupational, nonperinatal exposure is generally low. Transmission risk is modified by factors related to the source and extent of exposure. Determination of the HIV infection status of the exposure source may not be possible, and data on transmission risk by exposure type may not exist. Except in the setting of perinatal transmission, no studies have demonstrated the safety and efficacy of postexposure use of antiretroviral drugs for the prevention of HIV transmission in nonoccupational settings. Antiretroviral therapy used for PEP is associated with significant toxicity. The decision to initiate prophylaxis needs to be made in consultation with the patient, the family, and a clinician with experience in treatment of persons with HIV infection. If instituted, therapy should be started as soon as possible after an exposure-no later than 72 hours-and continued for 28 days. Many clinicians would use 3 drugs for PEP regimens, although 2 drugs may be considered in certain circumstances. Instruction for avoiding secondary transmission should be given. Careful follow-up is needed for psychologic support, encouragement of medication adherence, toxicity monitoring, and serial HIV antibody testing. PMID- 12777575 TI - It's wise to circumcise: time to change policy. PMID- 12777576 TI - Home visiting. PMID- 12777577 TI - A new look at meningomyeloceles. PMID- 12777578 TI - Errors in medical interpretation: our concerns for public health and a call for caution. PMID- 12777580 TI - Impact of the thimerosal controversy on hepatitis B vaccine coverage of infants born to women of unknown hepatitis B surface antigen status in Michigan. AB - OBJECTIVE: Hepatitis B vaccine is recommended for all infants, and the series may be started during the delivery admission. For infants who are born either to women who are positive for hepatitis B surface antigen (HBsAg) or to women whose HBsAg status is unknown, vaccination should be started within 12 hours of birth to prevent perinatal and early childhood hepatitis B virus infection. Because of concerns about mercury exposures from vaccines that contain thimerosal, the United States Public Health Service (USPHS) and the American Academy of Pediatrics (AAP) recommended in July 1999 that the first dose of hepatitis B vaccine be deferred until 2-6 months of age but only for infants who are born to HBsAg-negative women. To assess the impact on birth-dose vaccine coverage for infants who are born to women with unknown HBsAg status, we measured coverage before and after July 1999. METHODS: A sample of Michigan infants who were born to women whose HBsAg status was either unknown or missing were identified by reviewing newborn screening cards for infants who were born during 1) March-April 1999 (before recommendation changes [T1]); 2) July 15-September 15, 1999 (immediately after recommendation changes [T2]); and 3) March-April 2000 (6 months after resumption of pre-1999 practices were recommended [T3]). We verified maternal HBsAg screening and newborn hepatitis B vaccination by reviewing infant and maternal hospital records. RESULTS: Of 1201 infants who were born to women whose HBsAg status was indicated as unknown or missing on the newborn screening card during the 3 time periods, 216 (18%) were born to women whose status was truly unknown at the time of delivery, as determined by medical record review. During T1, 53% of these 216 infants received hepatitis B vaccine before hospital discharge, compared with 7% of infants who were born during T2 and 57% of infants who were born during T3. During T1, 19% of these infants received hepatitis B vaccine within 12 hours of birth compared with 1% of infants who were born during T2 and 14% of infants who were born during T3. CONCLUSIONS: Hepatitis B vaccine birth-dose coverage for infants who were born to women whose HBsAg status was unknown at the time of delivery was already low in Michigan before the July 1999 USPHS/AAP Joint Statement but decreased significantly during the 2 months after the USPHS/AAP Joint Statement. Abrupt changes in established vaccination recommendations for lower risk children may lead to decreased coverage among higher risk children. Increases in hepatitis B vaccine coverage at birth are necessary to reduce the risk of perinatal infection for infants who are born to women with unknown HBsAg status. PMID- 12777579 TI - Response to immunization with measles, tetanus, and Haemophilus influenzae type b vaccines in children who have human immunodeficiency virus type 1 infection and are treated with highly active antiretroviral therapy. AB - OBJECTIVE: To assess the level of immunity to measles, tetanus, and Haemophilus influenzae type b (Hib) in previously immunized children who have human immunodeficiency virus (HIV) infection and were treated with highly active antiretroviral therapy (HAART) and to determine the response to reimmunization. METHODS: Retrospective review of clinical data from children who have HIV-1 infection and were treated with HAART. Children were included in the analysis when they had a history of immunizations before treatment with HAART; had specific immunoglobulin G levels to tetanus, measles, or Hib measured after starting HAART but before the receipt of additional immunizations; were reimmunized while on HAART; and had postimmunization immunoglobulin G levels available. RESULTS: Nineteen children (median age: 7 years; range: 3-14 years) who were treated with 3 to 5 drug HAART regimens for a median of 20 months (range: 8-37) met the criteria for at least 1 antigen and were included in this review. Fifteen (79%) of the 19 had plasma RNA levels <50 copies/mL. The median CD4% before HAART was 26% (range: 1-41) and at the time of immunization, 35% (range: 20-54). Before reimmunization, 1 (5%) of 18 children had detectable antibody levels to measles, 6 (35%) of 17 had detectable antibody levels to tetanus, and 14 (78%) of 18 had detectable antibody levels to Hib. After immunization, 15 (83%) of 18, 10 (90%) of 11, and 3 (75%) of 4 seroconverted to measles, tetanus, and Hib, respectively. Antibody levels remained detectable after 1 year in the majority of children tested. CONCLUSIONS: Consideration should be given to readministering childhood immunizations to children who have HIV infection and are treated successfully with combination antiretroviral therapy. PMID- 12777581 TI - The state of office-based interventions for youth tobacco use. AB - Tobacco use is a serious pediatric health issue as dependence begins during childhood or adolescence in the majority of tobacco users. Primary care settings provide tremendous opportunities for delivering tobacco treatment to young tobacco users. Although evidence-based practice guidelines for treating nicotine dependence in youths are not yet available, professional organizations and the current clinical practice guideline for adults provide recommendations based on expert opinion. This article reports on the current tobacco treatment practices of pediatric and family practice clinicians, discusses similarities and differences between adolescent and adult tobacco use, summarizes research efforts to date and current cutting-edge research that may ultimately help to inform and guide clinicians, and presents existing recommendations regarding treating tobacco use in youths. Finally, recommendations are made for the primary care clinician, professional organizations, and health care systems and policies. Pediatricians and other clinicians can and should play an important role in treating tobacco dependence in youths. PMID- 12777582 TI - Delayed hemolytic transfusion reaction/hyperhemolysis syndrome in children with sickle cell disease. AB - OBJECTIVE: Alloimmunization in patients with sickle cell disease (SCD) has a reported incidence of 5% to 36%. One complication of alloimmunization is delayed hemolytic transfusion reaction/hyperhemolysis (DHTR/H) syndrome, which has a reported incidence of 11%. In patients with SCD, clinical findings in DHTR/H syndrome occur approximately 1 week after the red blood cell (RBC) transfusion and include the onset of increased hemolysis associated with pain and profound anemia. The hemoglobin (Hb) often drops below pretransfusion levels. In many reported adult cases, the direct antiglobulin test (DAT) remains negative and no new alloantibody is detected as the cause for these transfusion reactions. To date, few pediatric cases have been reported with this phenomenon. The objective of this study was to describe the clinical and laboratory findings of a case series in children who had SCD and experienced a DHTR/H syndrome at our institution. METHODS: An 11-year retrospective chart review of patients with discharge diagnosis of SCD and transfusion reaction was performed. DHTR/H syndrome was defined as the abrupt onset of signs and symptoms of accelerated hemolysis evidenced by an unexplained fall in Hb, elevated lactic dehydrogenase, elevated bilirubin above baseline, and hemoglobinuria, all occurring between 4 and 10 days after an RBC transfusion. Patient characteristics, time from transfusion, symptoms, reported DAT, new autoantibody or alloantibody formation, laboratory abnormalities, and complications were recorded. Patients with acute transfusion reactions were excluded. RESULTS: We encountered 7 patients who developed 9 episodes of DHTR/H syndrome occurring 6 to 10 days after RBC transfusion. Each presented with fever and hemoglobinuria. All but 1 patient experienced pain initially ascribed to vaso-occlusive crisis. The DAT was positive in only 2 of the 9 episodes. The presenting Hb was lower than pretransfusion levels in 8 of the 9 events. Severe complications were observed after the onset of DHTR/H: acute chest syndrome, n = 3; pancreatitis, n = 1; congestive heart failure, n = 1; and acute renal failure, n = 1. CONCLUSIONS: DHTR/H syndrome occurs in pediatric SCD patients, typically 1 week posttransfusion, and presents with back, leg, or abdominal pain; fever; and hemoglobinuria that may mimic pain crisis. Hb is often lower than it was at the time of original transfusion, suggesting the hemolysis of the patient's own RBCs in addition to hemolysis of the transfused RBCs; a negative DAT and reticulocytopenia are often present. Severe complications including acute chest syndrome, congestive heart failure, pancreatitis, and acute renal failure were associated with DHTR/H syndrome in our patients. DHTR/H in the pediatric sickle cell population is a serious and potentially life-threatening complication of RBC transfusion. It is important to avoid additional transfusions in these patients, if possible, because these may exacerbate the hemolysis and worsen the degree of anemia. DHTR/H syndrome must be included in the differential of a patient who has SCD and vaso-occlusive crisis who has recently had a transfusion. PMID- 12777583 TI - Evaluating the American Academy of Pediatrics diagnostic standard for Streptococcus pyogenes pharyngitis: backup culture versus repeat rapid antigen testing. AB - OBJECTIVE: The American Academy of Pediatrics recommends that all negative rapid diagnostic tests for Streptococcus pyogenes pharyngitis be backed up by culture, which creates a dilemma for clinicians who must make treatment decisions without complete diagnostic information at the time of visit. The use of a follow-up serial rapid antigen test instead of a follow-up culture would provide a more timely result. METHODS: Two swabs were collected from children who were suspected of having S pyogenes pharyngitis. Each swab was used for a culture and an OSOM Ultra Strep A Test rapid antigen test. The gold standard of comparison was defined as the identification of S pyogenes on either of the 2 culture plates. Three diagnostic strategies were evaluated: a single rapid antigen test, a rapid antigen test with follow-up rapid antigen test (rapid-rapid), and a rapid antigen test with follow-up culture (rapid-culture). RESULTS: A total of 210 (23.7%) of 887 throat cultures with matched data were identified with S pyogenes. A single rapid antigen test had a sensitivity of 87.6% (95% confidence interval [CI]: 83.2%-92.1%), the sensitivity of the rapid-rapid follow-up was 91.4% (95% CI: 87.6%-95.2%), and the sensitivity of the rapid-culture follow-up was 95.7% (95% CI: 93.0%-98.5%), which was significantly higher than the others. As shown in Fig 1, when these test strategies were evaluated on a subgroup with clinical symptoms commonly associated with S pyogenes pharyngitis, the sensitivities all increased and were no longer significantly different. None of the strategies reliably exceeded a 95% sensitivity threshold. CONCLUSIONS: The American Academy of Pediatrics strategy for S pyogenes detection in children with pharyngitis, requiring a backup culture for those with negative antigen tests, was not exceeded by any other test strategy; however, a rapid-rapid diagnostic strategy may approximate it with the use of judicious clinical selection of patients. PMID- 12777584 TI - Neurodevelopmental outcome of infants supported with extracorporeal membrane oxygenation after cardiac surgery. AB - OBJECTIVES: To evaluate the long-term neurodevelopmental outcome of infants who underwent cardiac surgery and required extracorporeal membrane oxygenation (ECMO) support, and to examine variables that predict death or disability in these patients. METHODS: We studied all infants who had congenital heart disease and were supported postoperatively with ECMO from 1990 to 2001 at our institution (n = 53). Medical records were reviewed retrospectively to obtain clinical variables. Neurologic and age-appropriate developmental examinations occurred at ages 1, 1.5, 2.5, and 4.5 years. Median age at follow-up was 55 months (9-101). Cognitive outcome was defined as suspect when scores were between 1 and 2 SD below the mean for age and abnormal when scores were >2 SD below mean for age. Neuromotor outcome was defined as suspect when the patient manifested clumsiness, tremor, or mild tone and reflex changes without functional limitations, and abnormal when there were functional limitations. RESULTS: In-hospital survival was 17 (32%) of 53. Of survivors, 14 (88%) of 16 are living and 1 patient was lost to follow-up. Of the 53 patients, 7 survived completely intact (13%). Seven (50%) of 14 patients had a normal cognitive outcome, 3 (21%) had a suspect cognitive outcome, and 4 (29%) were abnormal. Ten (72%) of 14 patients had a normal neuromotor outcome, 1 (7%) patient had a suspect neuromotor outcome, and 3 (21%) were abnormal. No survivor with an aortic cross-clamp time >40 minutes had a normal cognitive outcome. Nonsurvivors were more likely than survivors to have had cardiac arrest as an indication for ECMO (31% vs 6%), to have had a longer aortic cross-clamp time (mean 73 minutes vs 32 minutes), and to have required continuous arteriovenous hemofiltration (78% vs 35%). The age and weight at cannulation, gender, cardiac diagnosis, interval from surgery to ECMO, cardiopulmonary bypass time, diagnosis of sepsis or mediastinitis, and duration of ECMO were not significantly associated with survival. CONCLUSIONS: Although mortality was 68% in infants who had congenital heart disease and were treated with ECMO postoperatively, of those who survive to hospital discharge, 75% have a normal neuromotor outcome and 50% have a normal cognitive outcome. These high rates of mortality and disability suggest that increased attention be paid to neuroprotection in these complex disorders. PMID- 12777585 TI - Perinatal outcomes in two dissimilar immigrant populations in the United States: a dual epidemiologic paradox. AB - OBJECTIVE: Previous studies have addressed perinatal outcomes in Hispanic, black, and white non-Hispanic women and demonstrated that although foreign-born Mexican American women have many demographic and socioeconomic risk factors, their rates of low birth weight (LBW) infants and infant mortality are similar to those of white women. This phenomenon has been termed an epidemiologic paradox. There have been no population-based studies on women of Asian Indian origin, a relatively new, highly educated, and affluent immigrant group that has been reported to have a high rate of LBW infants. The objective of this study was to define the sociodemographic risk profile and perinatal outcomes in women of Asian Indian birth and to compare these outcomes to foreign-born Mexican American and US-born black and white women. METHODS: The vital records for self-reported foreign-born Asian Indian (0.8%) and Mexican women (26.7%) and US-born black (31.2%) and white women (31.2%) were extracted from California's 1 622 324 births, 1995-1997. Sociodemographic risk profiles; the percentage of LBW, very low birth weight (VLBW), prematurity, and intrauterine growth retardation (less than third percentile); and percentage of fetal, neonatal, and postneonatal death rates were compared. Logistic models were used to estimate the importance of selected sociodemographic and medical factors to the prediction of LBW infants in each racial/ethnic group. RESULTS: When compared with whites, US-born blacks and foreign-born Mexican mothers were at increased risk for adverse perinatal outcomes on the basis of higher levels of inadequate prenatal care, teen births, Medi-Cal paid delivery, and lower levels of maternal and paternal education. Foreign-born Asian Indian mothers had good prenatal care, were rarely teenagers, had dramatically higher levels of both maternal and paternal education, and had the lowest percentage of deliveries paid for by Medi-Cal. Black infants had the highest rates of prematurity; intrauterine growth retardation; LBW; and fetal, neonatal, and postneonatal mortality. Paradoxically, despite their high-risk profile, Mexicans did not have elevated levels of LBW or neonatal mortality. Conversely, Asian Indian infants, although seemingly of low sociodemographic risk, had high levels of LBW, growth retardation, and fetal mortality. Logistic regression analysis of independent risk factors for giving birth to an LBW infant showed higher maternal education, early access to prenatal care, and having private insurance to be protective in white non-Hispanic and black but not in Asian Indian and Mexican-born women. CONCLUSIONS: Despite their high socioeconomic status and early entry into care, foreign-born Asian Indian women have a paradoxically higher incidence of LBW infants and fetal deaths when compared with US-born whites. Factors that protect from giving birth to an LBW infant in white women were not protective among Asian Indian women. Current knowledge regarding factors that confer a perinatal advantage or disadvantage is unable to explain this new epidemiologic paradox. These findings highlight the need for additional research into both epidemiologic and biological risk factors that determine perinatal outcomes. PMID- 12777586 TI - Rates of pediatric injuries by 3-month intervals for children 0 to 3 years of age. AB - OBJECTIVE: Mortality and morbidity data on childhood injury are used to construct developmentally appropriate intervention strategies and to guide pediatric anticipatory counseling on injury prevention topics. Effective anticipatory guidance depends on detailed injury data showing how risks change as children develop. Conventional age groupings may be too broad to show the relationship between children's development and their risk of various causes of injury. Previous studies revealed differences in overall rates and specific causes of injury by year of age. However, single year of age rates for children younger than 4 years may not reflect the variations in risk as a result of rapid developmental changes. This study was designed to analyze injury rates for children younger than 4 years by quarter-year intervals to determine more specifically the age period of highest risk for injury and for specific causes. METHODS: We used data from 1996-1998 California hospital discharges and death certificates to identify day of age and external cause of injury (E-code) for children younger than 4 years. The number of California residents for each day of age was estimated from US Census of estimates of California's population by year of age for the midpoints (1996-1998). Rates were calculated by 3-month intervals. We grouped the E-codes into major categories that would be particularly relevant for developmentally related risks of injury specific to young children. The categorization took into account physical, motor, behavioral, and cognitive developmental milestones of children 0 to 3 years. RESULTS: There were a total of 23,173 injuries; 636 resulted in death. The overall annual rate for children aged 0 to 3 years was 371/100,000. Beginning at age 3 to 5 months, the overall rate of injury rapidly increased with increased age, peaking at 15 to 17 months. The mean injury rate calculated for each single year of age did not reflect the variation and the highest rate of injury by quarter year of age for children younger than 1 year, 1 year, and 2 years. The leading major causes of injury in descending order were falls, poisoning, transportation, foreign body, and fires/burns. The overall rate of the major category of falls exceeded poisoning, the second leading cause of injury, by a factor of 2. Age-related differences were detected within each major cause of injury. For children 0 to 12 months of age, there was a different leading cause of specific injury for each 3-month period: other falls from height (0-2 months), battering (3-5 months), falls from furniture (6-8 months), and nonairway foreign body (9-11 months). Hot liquid and vapor injuries were the leading specific causes for children 12 to 17 months. Poisoning by medication was the leading specific cause of injury for all age groups from 18 to 35 months and exceeded poisoning by other substances. Pedestrian injury was the leading specific cause of injury for all age groups from 36 to 47 months. Fall from furniture has the highest rates of specific causes of falls from age 3 to 47 months. Fall from stairs peaked at age 6 to 8 months and 9 to 11 months. Fall from buildings was highest at 24 to 26 months. Poisoning by medication peaked at age 21 to 23 months, but poisoning by other substances peaked at 15 to 17 months. The motor vehicle occupant injury rates were fairly stable over the age span of this study. The pedestrian injury rate increased beginning at age 12 to 14 months and by 15 to 17 months was double that of the motor vehicle occupant. Foreign body had a marked peak at age 9 to 11 months. Both battering and neglect rates were highest among infants 0 to 2 and 3 to 5 months. Bathtub submersions had a narrow peak at age 6 to 11 months. Other submersions peaked at 12 to 14 months and remained high until 33 to 35 months. CONCLUSIONS: We departed from usual groupings of E-codes and devised groupings that would be reflective of age related developmental characteristics. Differences in rates by narrow age groups for young children can be related to developmental achievements, w can be related to developmental achievements, which place the child at risk for specific causes of injury. We found marked variability in both rates and leading causes of injury by 3-month interval age groupings that were masked by year of age analyses. Children aged 15 to 17 months had the highest overall injury rate before age 15 years. This coincides with developmental achievements such as independent mobility, exploratory behavior, and hand-to-mouth activity. The child is able to access hazards but has not yet developed cognitive hazard awareness and avoidance skills. A remarkable finding was the high rate of battering injury among infants 0 to 5 months, suggesting the need to address potential child maltreatment in the perinatal period. Poisoning was the second major leading cause of injury; more than two thirds were medication. Cultural factors may influence views of medications, storage practices, use of poison control system telephone advice, and risk of toddler poisoning. The pedestrian injury rate doubled between 12 and 14 months and 15 and 17 months and exceeded motor vehicle occupant injury rates for each 3-month interval from 15 to 47 months. Pedestrian injury has not received sufficient attention in general and certainly not in injury prevention counseling for children younger than 4 years. Anticipatory guidance for pedestrian injury should be incorporated before 1 year of age. Effective strategies must be based on the epidemiology of childhood injury. Pediatricians and other pediatric health care providers are in a unique position to render injury prevention services to their patients. Integrating injury prevention messages in the context of developmental assessments of the child is 1 strategy. These data can also be used for complementary childhood injury prevention strategies such as early intervention programs for high-risk families for child abuse and neglect, media and advocacy campaigns, public policies, and environmental and product design. PMID- 12777587 TI - Which psychiatric patients board on the medical service? AB - OBJECTIVES: Patients who require psychiatric hospitalization may be admitted to a medical service only because there are no available inpatient psychiatric beds. These patients are psychiatric "boarders." The goals of this study were to describe the extent of the boarder problem and to compare the characteristics of patients who are placed successfully into psychiatric facilities from the emergency department (ED) with those who require admission to the medical service as a boarder. METHODS: A retrospective cohort study of a large pediatric ED was conducted. Included were patients who required inpatient psychiatric admission between July 1, 1999, and June 30, 2000. Patients were excluded when they needed inpatient medical treatment before psychiatric placement. The main outcome measured was placement into a psychiatric facility or boarding on medical service. RESULTS: Of the 315 patients who presented to the ED and required psychiatric admission, 103 (33%) were boarded on the medical service. Multivariate logistic regression demonstrated an increased odds of boarding for age 10 to 13 years (adjusted odds ratio [AOR]: 3.5; 95% confidence interval [CI]: 1.8-6.6), black race (AOR: 2.3; 95% CI: 1.1-4.8), presenting on a weekend or holiday (AOR: 3.8; 95% CI: 1.6-8.8), and presenting from October to June (October December 1999 [AOR: 4.7; 95% CI: 1.7-13.4], January-March 2000 [AOR: 14.5; 95% CI: 4.9-42.6], and April-June 2000 [AOR: 10.4; 95% CI: 3.5-30.2]) but a decreased odds for 1 insurance company (AOR: 0.08; 95% CI: 0.02-0.4). There was a linear increase in odds of boarding as severity of homicidal ideation increased from none to mild (AOR: 1.5; 95% CI: 1.2-1.8) to moderate (AOR: 2.3; 95% CI: 2.0-2.6) to severe (AOR: 3.5; 95% CI: 3.2-3.8). Suicidal patients also had increased risk of boarding (AOR: 2.2; 95% CI: 1.2-4.3). CONCLUSIONS: Boarders are a problem in pediatrics, and this study identifies multiple characteristics that were associated with increasing a youth's odds of becoming a boarder at this institution. The suicidal and homicidal symptom results suggest a reverse triage system in which sicker patients are not necessarily given priority by psychiatric facilities. These data highlight mental health practices that need to be reassessed to ensure optimal care for youths with acute mental illness. PMID- 12777588 TI - Substance use problems and associated psychiatric symptoms among adolescents in primary care. AB - OBJECTIVE: Substance use disorders (SUDs) are associated with other mental disorders in adolescence, but it is unclear whether less severe substance use problems (SUPs) also increase risk. Because youths with SUPs are most likely to present first to their site of primary care, it is important to establish the presence and patterns of psychiatric comorbidity among adolescent primary care patients with subdiagnostic use of alcohol or other drugs. The objective of this study was to determine the association between level of substance use and psychiatric symptoms among adolescents in a primary care setting. METHODS: Patients who were aged 14 to 18 years and receiving routine care at a hospital based adolescent clinic were eligible. Participants completed the Problem Oriented Screening Instrument for Teenagers Substance Use/Abuse scale, which is designed to detect social and legal problems associated with alcohol and other drugs, and the Adolescent Diagnostic Interview, which evaluates for Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition diagnoses of substance abuse/dependence and 8 types of psychiatric symptoms. We examined gender-specific associations of no/nonproblematic substance use (NSU), SUP, and SUD with psychiatric symptom presence (any symptoms within each type), score (symptom scores summed across all types), and number of types (number of different symptom types endorsed). RESULTS: Of 538 adolescents (68% female; mean +/- standard deviation age: 16.6 +/- 1.4 years), 66% were classified with NSU, 18% with SUP, and 16% with SUD, and 80% reported having at least 1 type of psychiatric symptom in the previous 12 months. Symptoms of anxiety were most common (60% of both boys and girls), followed by symptoms of depression among girls (51%) and symptoms of attention-deficit disorder (ADD) among boys (47%). Compared with those with NSU, youths with SUP and those with SUD were more likely to report symptom presence for several types of psychiatric symptoms. Girls with SUP or SUD had increased odds of reporting symptoms of mania, ADD, and conduct disorder; girls with SUD were at increased risk for symptoms of depression, eating disorders, and hallucinations or delusions. Boys with SUP had increased odds of ADD symptoms, whereas boys with SUD had increased odds of reporting hallucinations or delusions. Boys with SUP or SUD had increased odds of reporting symptoms of conduct disorder. Youths with SUP and SUD also had higher psychiatric symptom scores and reported a wider range of psychiatric symptom types (number of types) compared with youths with NSU. CONCLUSIONS: Like those with SUD, adolescents with subdiagnostic SUP were at increased risk for experiencing a greater number of psychiatric symptoms and a wider range of psychiatric symptom types than youths with NSU. Specifically, adolescents with SUP are at increased risk for symptoms of mood (girls) and disruptive behavior disorders (girls and boys). These findings suggest the clinical importance of SUP and support the concept of a continuum between subthreshold and diagnostic substance use among adolescents in primary care. Identification of youths with SUP may allow for intervention before either the substance use or any associated psychiatric problems progress to more severe levels. PMID- 12777589 TI - Long-term safety of fluticasone propionate and nedocromil sodium on bone in children with asthma. AB - OBJECTIVE: Inhaled corticosteroids are recommended as first-line therapy for pediatric asthma. However, few controlled long-term studies have investigated their effect on bone mineral density (BMD) and growth. METHODS: Children who were aged 6 to 14 years and had persistent asthma were randomized to 24 months' treatment with fluticasone propionate (FP) 200 micro g/d or nedocromil sodium (NS) 8 mg/d (if uncontrolled, maximum doses of 400 micro g/d and 16 mg/d, respectively). BMD was assessed blind and analyzed at a central facility on the basis of dual-energy x-ray absorptiometry measurements of the lumbar spine and femoral neck at months 0, 6, 12, and 24. Height was measured at months 0, 12, and 24. Efficacy parameters (lung function, asthma control, occurrence of exacerbations) were measured every 3 months. RESULTS: In total, 174 children were randomized to treatment (87 received FP, and 87 received NS). At month 24, the adjusted mean percentage increase in lumbar spine BMD was 11.6% in the FP group compared with 10.4% in NS-treated children (95% confidence interval for treatment difference: -0.7% to 3.1%). The corresponding increases in femoral neck BMD were 8.9% and 8.5%, respectively. There was no significant difference in growth between the 2 groups: adjusted mean growth rates were 6.1 cm/y with FP and 5.8 cm/y with NS. FP was significantly superior for every efficacy parameter investigated and was similarly well tolerated as NS. CONCLUSIONS: The long-term effects of FP and NS on BMD accrual and growth are similar among children with asthma. The benefit:risk ratio of FP may be considered superior to that of NS. PMID- 12777590 TI - Fluoroquinolone safety in pediatric patients: a prospective, multicenter, comparative cohort study in France. AB - OBJECTIVE: To evaluate the safety of fluoroquinolones (FQ) in comparison with other antibiotics in pediatric patients. METHODS: A multicenter, observational, comparative cohort study was conducted between 1998 and 2000 in French pediatric departments. Patients who were receiving systemic FQ were included and matched to control patients who were receiving other antibiotics. Antibiotic-associated potential adverse events (PAEs) were recorded prospectively in both groups, and their rates were compared using univariate and multivariate analyses. RESULTS: Patients were recruited from 73 centers: 276 patients were exposed to FQ, and 249 composed the control group. Among patients who were exposed to FQ, 23% were younger than 2 years, 33% had cystic fibrosis, and PAEs occurred in 52 patients, leading to withdrawal for 11. The odds ratio for PAE in the FQ group was 3.7 (95% confidence interval: 1.9-7.5) and was not significantly modified after adjustment for potential confounders. Musculoskeletal PAEs also occurred more frequently in the FQ group (3.8%) than in controls (0.4%); they were recorded in 10 patients who were receiving standard FQ doses and were of moderate intensity and transient. CONCLUSION: The rates of PAEs and musculoskeletal PAEs were higher for the FQ group than the control group. This observation supports the American Academy of Pediatrics statement restricting off-label FQ use in pediatric patients to second-line treatment in a limited number of situations. PMID- 12777591 TI - Rest-activity patterns in children with hypopituitarism. AB - OBJECTIVES: Physical lesions in the region of the suprachiasmatic nuclei, which are the site of a circadian clock, result in abnormal circadian rhythmicity in animals, yet the extent of biological rhythm problems in individuals with anatomic or functional lesions in the hypothalamic-pituitary region are largely unknown. To address this issue, we examined patterns of rest and activity of children with hypopituitarism. METHODS: Children who were between the ages of 2 and 18 years and had the diagnosis of panhypopituitarism were evaluated. Twenty children were studied, including children with septo-optic dysplasia (SOD), congenital hypopituitarism, brain tumors, closed head trauma, and head irradiation. For assessing patterns of activity, individuals wore Actiwatches for 3-4 weeks to measure patterns of gross motor activity. RESULTS: Seventeen children had normal patterns of rest and activity, with an average period length of 24.01 +/- 0.01 hours. Three children, including 2 with SOD and 1 with a hypothalamic germinoma, showed abnormal activity patterns in which there was not consolidated rest at night. One patient with an optic glioma had nonentrained circadian phase. CONCLUSIONS: A proportion of children with hypopituitarism have abnormal daily rest-activity patterns. Children with anterior hypothalamic tumors and SOD seem to be at risk for circadian system dysfunction. PMID- 12777592 TI - Symptoms of posttraumatic stress disorder in parents of transplant recipients: incidence, severity, and related factors. AB - OBJECTIVE: To examine the incidence, severity, and factors related to posttraumatic stress disorder (PTSD) symptoms in parents of pediatric solid organ transplant recipients. METHOD: A total of 170 caregivers of pediatric transplant recipients completed self-report measures of psychological functioning between 10 and 38 months after their child's most recent transplant. Demographic data, child health variables, and ratings of medical attitudes and social functioning were also collected to help explain individual differences in psychological functioning. RESULTS: Although caregivers of pediatric transplant recipients did not report elevated levels of depression or anxiety, they did report elevated levels of PTSD symptoms. Multiple regression analyses revealed that PTSD symptoms were most strongly associated with parent reports of child health, family impact of the transplant, and attitudes toward medical caregivers. CONCLUSIONS: PTSD seems to be relatively common in parents of pediatric transplant recipients and may be largely the result of how parents perceive and interpret the transplant experience. PMID- 12777593 TI - Strangulation with intravenous tubing: a previously undescribed adverse advent in children. AB - Nonintentional strangulation in children is a widely recognized risk as a result of the vulnerability of their airway to occlusion by relatively low pressures. We describe 2 cases of strangulation by intravenous (IV) tubing in infants, 1 of which was fatal. This is the first documentation in the health science literature of this as a potential adverse consequence of IV therapy in young children. It is important that hospitals that care for such children recognize this potential risk and implement the appropriate strategies to minimize or eliminate it. Preventive interventions may include ongoing assessment of the need for continuous rather than intermittent IV infusions (saline or heparin locked IV sites), individualized level of supervision according to the child's age and behavior, and engineering modifications to the IV equipment. PMID- 12777595 TI - Family pediatrics: report of the Task Force on the Family. AB - WHY A TASK FORCE ON THE FAMILY? The practice of pediatrics is unique among medical specialties in many ways, among which is the nearly certain presence of a parent when health care services are provided for the patient. Regardless of whether parents or other family members are physically present, their influence is pervasive. Families are the most central and enduring influence in children's lives. Parents are also central in pediatric care. The health and well-being of children are inextricably linked to their parents' physical, emotional and social health, social circumstances, and child-rearing practices. The rising incidence of behavior problems among children attests to some families' inability to cope with the increasing stresses they are experiencing and their need for assistance. When a family's distress finds its voice in a child's symptoms, pediatricians are often parents' first source for help. There is enormous diversity among families diversity in the composition of families, in their ethnic and racial heritage, in their religious and spiritual orientation, in how they communicate, in the time they spend together, in their commitment to individual family members, in their connections to their community, in their experiences, and in their ability to adapt to stress. Within families, individuals are different from one another as well. Pediatricians are especially sensitive to differences among children-in their temperaments and personalities, in their innate and learned abilities, and in how they view themselves and respond to the world around them. It is remarkable and a testament to the effort of parents and to the resilience of children that most families function well and most children succeed in life. Family life in the United States has been subjected to extensive scrutiny and frequent commentary, yet even when those activities have been informed by research, they tend to be influenced by personal experience within families and by individual and cultural beliefs about how society and family life ought to be. The process of formulating recommendations for pediatric practice, public policy, professional education, and research requires reaching consensus on some core values and principles about family life and family functioning as they affect children, knowing that some philosophic disagreements will remain unresolved. The growing multicultural character of the country will likely heighten awareness of our diversity. Many characteristics of families have changed during the past 3 to 5 decades. Families without children younger than 18 years have increased substantially, and they are now the majority. The average age at marriage has increased, and a greater proportion of births is occurring to women older than 30 years. Between 1970 and 2000, the proportion of children in 2-parent families decreased from 85% to 69%, and more than one quarter (26%) of all children live with a single parent, usually their mother. Most of this change reflects a dramatic increase in the rate of births to unmarried women that went from 5.3% in 1960 to 33.2% in 2000. Another factor in this change is a slowly decreasing but still high divorce rate that is roughly double what it was in the mid-1950s. Family income is strongly related to children's health, and the financial resources that families have available are closely tied to changes in family structure. Family income in real dollars has trended up for many decades, but the benefits have not been shared equally. For example, the median income of families with married parents has increased by 146% since 1970, but female-headed households have experienced a growth of 131%. More striking is that in 2000, the median income of female-headed households was only 47% of that of married-couple families and only 65% of that of families with 2 married parents in which the wife was not employed. Not surprising, the proportion of children who live in poverty is approximately 5 times greater for female-headed families than for married-couple families. The comped families than for married-couple families. The composition of children's families and the time parents have for their children affect child rearing. Consequent to the increase in female-headed households, rising economic and personal need, and increased opportunities for women, the proportion of mothers who are in the workforce has climbed steadily over the past several decades. Currently, approximately two thirds of all mothers with children younger than 18 years are employed. Most families with young children depend on child care, and most child care is not of good quality. Reliance on child care involves longer days for children and families, the stress imposed by schedules and created by transitions, exposure to infections, and considerable cost. An increasing number and proportion of parents are also devoting time previously available to their children to the care of their own parents. The so-called "sandwich generation" of parents is being pulled in multiple directions. The amount and use of family time also has changed with a lengthening workday, including the amount of commuting time necessary to travel between work and home, and with the intrusion of television and computers into family life. In public opinion polls, most parents report that they believe it is more difficult to be a parent now than it used to be; people seem to feel more isolated, social and media pressures on and enticements of their children seem greater, and the world seems to be a more dangerous place. Social and public policy has not kept up with these changes, leaving families stretched for time and stressed to cope and meet their responsibilities. What can and what should pediatrics do to help families raise healthy and well-adjusted children? How can individual pediatricians better support families? FAMILY PEDIATRICS: The American Academy of Pediatrics (AAP) Board of Directors appointed the Task Force on the Family to help guide the development of public policy and recommend how to assist pediatricians to promote well-functioning families (see Appendix). The magnitude of the assigned work required task force members to learn a great deal from research and researchers in the fields of social and behavioral sciences. A review of some critical literature was completed by a consultant to the task force and accompanies this report. That review identified a convergence of pediatrics and research on families by other disciplines. The task force found that a great deal is known about family functioning and family circumstances that affect children. With this knowledge, it is possible to provide pediatric care in a way that promotes successful families and good outcomes for children. The task force refers to that type of care as "family-oriented care" or "family pediatrics" and strongly endorses policies and practices that promote the adoption of this 2-generational approach as a hallmark of pediatrics. During the past decade, family advocates have successfully promoted family-centered care, "the philosophies, principles and practices that put the family at the heart or center of services; the family as the driving force." Most pediatricians report that they involve families in the decision making regarding the health care of their child and make an effort to understand the needs of the family as well as the child. Family pediatrics, like family-centered care, requires an active, productive partnership between the pediatrician and the family. But family pediatrics extends the responsibilities of the pediatrician to include screening, assessment, and referral of parents for physical, emotional, or social problems or health risk behaviors that can adversely affect the health and emotional or social well-being of their child. FAMILY CONTEXT OF CHILD HEALTH: The power and importance of families to children arises out of the extended duration for which children are dependent on adults to meet their basic needs. Children's needs for which only a family can provide include social support, socialization, and coping and life skills. Their self-esteem grows from being cared for, loved, and valued and feeling that they are part of a social unit that shares values, communicates openly, and provides companionship. Families transmit and interpret values to their children and often serve as children's connection to the larger world, especially during the early years of life. Although schools provide formal education, families teach children how to get along in the world. Often, efforts to discuss families and make recommendations regarding practice or policy stumble over disagreements about the definition of a family. The task force recognized the diversity of families and chose not to operate from the position of a fixed definition. Rather, the task force, which was to address pediatrics, decided to frame its deliberations and recommendations around the functions of families and how various aspects of the family context influence child rearing and child health. One model of family functioning that implicitly guided the task force is the family stress model (Fig 1). Stress of various sorts (eg, financial or health problems, lack of social support, unhappiness at work, unfortunate life events) can cause parents emotional distress and cause couples conflict and difficulty with their relationship. These responses to stress then disrupt parenting and the interactions between parent and child and can lead to short-term or lasting poor outcomes. The earlier these events transpire and the longer that the disruption lasts, the worse the outcomes for children. The task force favors efforts to encourage and support marriage yet recognizes that every family constellation can produce good outcomes for children and that none is certain to yield bad ones. (ABSTRACT TRUNCATED) PMID- 12777596 TI - Converging trends in family research and pediatrics: recent findings for the American Academy of Pediatrics Task Force on the Family. PMID- 12777597 TI - Proceedings of a symposium on pediatric food allergy. April 20, 2002. AB - Food allergy seems to be increasing in prevalence, significantly decreases the quality of life for patients and their families, and has become a common diagnostic and management issue for the pediatrician. Studies now a decade old showed that 6% to 8% of children younger than 3 years experience documented adverse reactions to foods. Several studies have defined the prevalence of allergy to specific foods in childhood. Population-based studies document a prevalence of cow milk allergy in 1.9% to 3.2% of infants and young children, egg allergy in 2.6% of children by age 2.5 years, and peanut allergy in 0.4% to 0.6% of those younger than 18 years. Overall, the typical allergens of infancy and early childhood are egg, milk, peanut, wheat, and soy, whereas allergens that are responsible for severe reactions in older children and adults are primarily caused by peanut, tree nuts, and seafood. Allergy to fruits and vegetables are prominent but usually not severe. For diagnostic purposes, it is instructive to consider the prevalence of food allergy as a cause of specific disorders. For example, food allergy accounts for 20% of acute urticaria, is present in 37% of children with moderate to severe atopic dermatitis and approximately 5% with atopic asthma, and is the most frequent cause of anaphylaxis outside the hospital setting. PMID- 12777598 TI - Mucosal immunity. AB - Food allergy is the manifestation of an abnormal immune response to antigen delivered by the oral route. Normal mucosal immune responses are generally associated with suppression of immunity. A normal mucosal immune response relies heavily on a number of factors: strong physical barriers, luminal digestion of potential antigens, selective antigen sampling sites, and unique T-cell subpopulations that effect suppression. In the newborn, several of these pathways are not matured, allowing for sensitization rather than suppression. With age, the mucosa associated lymphoid tissue matures, and in most individuals this allows for generation of the normal suppressed tone of the mucosa associated lymphoid tissue. As a consequence, food allergies are largely outgrown. This article deals with the normal facets of mucosal immune responses and postulates how the different processes may be defective in food-allergic patients. PMID- 12777599 TI - Anaphylaxis and emergency treatment. AB - Food anaphylaxis is now the leading known cause of anaphylactic reactions treated in emergency departments in the United States. It is estimated that there are 30 000 anaphylactic reactions to foods treated in emergency departments and 150 to 200 deaths each year. Peanuts, tree nuts, fish, and shellfish account for most severe food anaphylactic reactions. Although clearly a form of immunoglobulin E mediated hypersensitivity, the mechanistic details responsible for symptoms of food-induced anaphylaxis are not completely understood, and in some cases, symptoms are not seen unless the patient exercises within a few hours of the ingestion. At the present time, the mainstays of therapy include educating patients and their caregivers to strictly avoid food allergens, to recognize early symptoms of anaphylaxis, and to self-administer injectable epinephrine. However, clinical trials are now under way for the treatment of patients with peanut anaphylaxis using recombinant humanized anti-immunoglobulin E antibody therapy, and novel immunomodulatory therapies are being tested in animal models of peanut-induced anaphylaxis. PMID- 12777600 TI - Clinical aspects of gastrointestinal food allergy in childhood. AB - Gastrointestinal food allergies are a spectrum of disorders that result from adverse immune responses to dietary antigens. The named disorders include immediate gastrointestinal hypersensitivity (anaphylaxis), oral allergy syndrome, allergic eosinophilic esophagitis, gastritis, and gastroenterocolitis; dietary protein enterocolitis, proctitis, and enteropathy; and celiac disease. Additional disorders sometimes attributed to food allergy include colic, gastroesophageal reflux, and constipation. The pediatrician faces several challenges in dealing with these disorders because diagnosis requires differentiating allergic disorders from many other causes of similar symptoms, and therapy requires identification of causal foods, application of therapeutic diets and/or medications, and monitoring for resolution of these disorders. This review catalogs the spectrum of gastrointestinal food allergies that affect children and provides a framework for a rational approach to diagnosis and management. PMID- 12777601 TI - Skin manifestations of food allergy. AB - The pediatrician is faced with evaluating a panoply of skin rashes, a subset of which may be induced by food allergy. Acute urticaria is a common manifestation of an allergic skin response to food, but food is rarely a cause of chronic urticaria. Approximately one third of infants/children with moderate to severe atopic dermatitis have food allergy. Although diagnosis of acute urticaria provoked by a food may be evident from a straightforward history and confirmed by diagnostic tests to detect food-specific IgE antibody, determination of the role of food allergy in patients with atopic dermatitis is more difficult and may require additional diagnostic maneuvers, including elimination diets and oral food challenges. The immunopathologic basis of food-allergic disorders that affect the skin and a rational approach to diagnosis and treatment are discussed. Additional disorders that are caused by or mimic ones caused by food allergy are reviewed. PMID- 12777602 TI - Respiratory manifestations of food allergy. AB - Food allergy may present with a variety of respiratory tract symptoms that generally involve immunoglobulin E antibody-mediated responses. Exposure is typically through ingestion, but in some cases, inhalation of airborne food particles may trigger these reactions. Upper and lower respiratory tract reactions are often a significant component of multisystem, anaphylactic reactions. However, chronic or isolated asthma or rhinitis induced by food is unusual. It is important to recognize that food allergy in early childhood is a marker indicating an increased risk to develop respiratory allergy. The role of food allergy in otitis media is controversial and probably is extremely rare. Likewise, asthmatic responses to food additives can occur but are uncommon. Studies using blinded oral food challenges have demonstrated that foods can elicit airway hyperreactivity and asthmatic responses. Therefore, an evaluation for food allergy should be considered in patients who are at risk, including those with recalcitrant or otherwise unexplained acute, severe asthma exacerbations, asthma triggered after ingestion of particular foods, and asthma that is accompanied by other manifestations of food allergy (e.g., anaphylaxis, moderate to severe atopic dermatitis). PMID- 12777603 TI - The natural history of food allergy. AB - The natural history of food allergy refers to the development of food sensitivities as well as the possible loss of the same food sensitivities over time. Most food allergy is acquired in the first 1 to 2 years of life, whereas the loss of food allergy is a far more variable process, depending on both the individual child and the specific food allergy. For example, whereas most milk allergy is outgrown over time, most allergies to peanuts and tree nuts are never lost. In addition, whereas some children may lose their milk allergy in a matter of months, the process may take as long as 8 or 10 years in other children. This review provides an overview of the natural history of food allergy and provides specific information on the natural course of the most common childhood food allergies. PMID- 12777604 TI - Diagnostic evaluation. AB - The diagnosis and management of adverse food reactions is a challenge for pediatricians and other primary care providers. Children of all ages may present with a variety of symptoms that parents have attributed to constituents of the diet. An approach has been devised to aid pediatricians in the evaluation of these children. The process begins with a detailed history. This history may be used to classify the problem into categories of symptoms and possible mechanisms. When common food offenders are suspected of causing symptoms, it is reasonable to obtain in vitro tests that may then be used to decide whether elimination of certain foods is indicated and whether a referral to an allergist is needed. Using this approach, the physician may be able to identify children who are experiencing food-allergic symptoms and aid frustrated families in dealing with problems that have not had apparent solutions. The application of specific testing and the assessment of the results are discussed. Also reviewed are the techniques used by allergists so that pediatricians may help families understand procedures that will be recommended and performed. PMID- 12777605 TI - Nutritional management of pediatric food hypersensitivity. AB - The diagnosis and management of food allergy requires attention to several important dietary issues. Successful exclusion of identified dietary allergens requires extensive education regarding the interpretation of ingredient labels of commercial products and an appreciation for issues of cross-contact in settings such as restaurants and commercial manufacturing. Once a food or food group is eliminated, attention must be focused on potential dietary insufficiencies resulting from these exclusions. These dietary issues are also central to the successful use of diagnostic elimination diets and physician-supervised oral food challenges. This review provides a framework for the dietary management of food hypersensitivity in infants and children both for short-term diagnostic and long term therapeutic purposes. In addition, approaches for maternal dietary restriction for breastfed infants with food allergy and the introduction of solid foods to atopic infants are reviewed. PMID- 12777606 TI - Daily coping strategies for patients and their families. AB - The diagnosis of food allergy in a child has an impact on every minute of every day for the child and the child's family. The patient and family must learn how to read labels, adapt recipes, and educate other family members, child care providers, camp counselors, and teachers. They must know how to recognize symptoms of a reaction and what to do during a reaction. Decisions such as which restaurant to go to and where to go on vacation take on new meaning, as family decisions must be centered on avoidance of the child's food allergen. It is possible to manage food allergies successfully while allowing the child to participate in common childhood activities. Education of the family is key. This discussion provides the pediatrician or primary care physician with practical information for educating patients and their families about managing food allergy. PMID- 12777607 TI - Food allergen avoidance in the prevention of food allergy in infants and children. AB - Food allergy afflicts an increasing number of infants and children and is associated with both clinical and familial burdens. To help lessen this burden, the Nutritional Committees from the American Academy of Pediatrics and jointly the European Society for Pediatric Allergology and Clinical Immunology and the European Society for Pediatric Gastroenterology, Hepatology, and Nutrition published recommendations to prevent and treat food allergy. Although there is much in common with these recommendations, differences exist. This review compares, contrasts, and reconciles them, presenting the evidence that has led to their statements. PMID- 12777608 TI - Future approaches to food allergy. AB - Food allergy affects approximately 2% of the general US population, and its prevalence seems to be increasing. Despite the potential for a fatal outcome, no definitive therapies are available for food allergy. This article reviews novel approaches for the diagnosis and treatment of food allergy. Improved diagnostic methods include more precise in vitro and in vivo tests for immunoglobulin E mediated food allergies, in vitro assays for predicting development of oral tolerance, and novel noninvasive tests for cell-mediated food allergies such as patch testing, cytokine assays, and detection of eosinophil activation markers. Several promising novel immunomodulatory approaches to food allergy are discussed, including monoclonal anti-immunoglobulin E; probiotics; traditional Chinese medicine; and immunotherapy with modified food proteins, peptides, bacterial adjuvants, and immunostimulatory sequences. PMID- 12777609 TI - The primary pediatrician's role when a death occurs in a family in one's practice. PMID- 12777610 TI - HIV infection in infants, children, and adolescents. PMID- 12777611 TI - Childhood malabsorption. PMID- 12777612 TI - Index of suspicion. PMID- 12777613 TI - Law and the child. PMID- 12777616 TI - Efficiency of single-nucleotide polymorphism haplotype estimation from pooled DNA. AB - The efficiency of single-nucleotide polymorphism haplotype analysis may be increased by DNA pooling, which can dramatically reduce the number of genotyping assays. We develop a method for obtaining maximum likelihood estimates of haplotype frequencies for different pool sizes, assess the accuracy of these estimates, and show that pooling DNA samples is efficient in estimating haplotype frequencies. Although pooling K individuals increases ambiguities, at least for small pool size K and small numbers of loci, the uncertainty of estimation increases Thr (I244T) mutation in the AGXT gene, with 14 of 16 patients homozygous for this mutation. Four polymorphisms within AGXT and regional microsatellites also were shared in their haplotypes (AGXT*LTM), consistent with a founder effect. The consequences of these amino acid changes were investigated. Although I244T alone did not affect AGXT activity or subcellular localization, when present in the same protein molecule as Leu-11 --> Pro (L11P), it resulted in loss of enzymatic activity in soluble cell extracts. Like its normal counterpart, the AGXT*LTM protein was present in the peroxisomes but it was insoluble in detergent-free buffers. The polymorphism L11P behaved as an intragenic modifier of the I244T mutation, with the resulting protein undergoing stable interaction with molecular chaperones and aggregation. This aggregation was temperature-sensitive. AGXT*LTM expressed in Escherichia coli, as a GST-fusion protein, and in insect cells could be purified and retained enzymatic activity. Among various chemical chaperones tested in cell culture, betaine substantially improved the solubility of the mutant protein and the enzymatic activity in cell lysates. In summary, I244T, the second most common mutation responsible for PH1, is a protein conformational disease that may benefit from new therapies with pharmacological chaperones or small molecules to minimize protein aggregation. PMID- 12777627 TI - Discovery of diverse thyroid hormone receptor antagonists by high-throughput docking. AB - Treatment of hyperthyroidism, a common clinical condition that can have serious manifestations in the elderly, has remained essentially unchanged for >30 years. Directly antagonizing the effect of the thyroid hormone at the receptor level may be a significant improvement for the treatment of hyperthyroid patients. We built a computer model of the thyroid hormone receptor (TR) ligand-binding domain in its predicted antagonist-bound conformation and used a virtual screening algorithm to select 100 TR antagonist candidates out of a library of >250,000 compounds. We were able to obtain 75 of the compounds selected in silico and studied their ability to act as antagonists by using cultured cells that express TR. Fourteen of these compounds were found to antagonize the effect of T3 on TR with IC50s ranging from 1.5 to 30 microM. A small virtual library of compounds, derived from the highest affinity antagonist (1-850) that could be rapidly synthesized, was generated. A second round of virtual screening identified new compounds with predicted increased antagonist activity. These second generation compounds were synthesized, and their ability to act as TR antagonists was confirmed by transfection and receptor binding experiments. The extreme structural diversity of the antagonist compounds shows how receptor-based virtual screening can identify diverse chemistries that comply with the structural rules of TR antagonism. PMID- 12777628 TI - Predicting survival in patients with metastatic kidney cancer by gene-expression profiling in the primary tumor. AB - To identify potential molecular determinants of tumor biology and possible clinical outcomes, global gene-expression patterns were analyzed in the primary tumors of patients with metastatic renal cell cancer by using cDNA microarrays. We used grossly dissected tumor masses that included tumor, blood vessels, connective tissue, and infiltrating immune cells to obtain a gene-expression "profile" from each primary tumor. Two patterns of gene expression were found within this uniformly staged patient population, which correlated with a significant difference in overall survival between the two patient groups. Subsets of genes most significantly associated with survival were defined, and vascular cell adhesion molecule-1 (VCAM-1) was the gene most predictive for survival. Therefore, despite the complex biological nature of metastatic cancer, basic clinical behavior as defined by survival may be determined by the gene expression patterns expressed within the compilation of primary gross tumor cells. We conclude that survival in patients with metastatic renal cell cancer can be correlated with the expression of various genes based solely on the expression profile in the primary kidney tumor. PMID- 12777629 TI - A novel social polymorphism in a primitively eusocial bee. AB - Halictine sweat bees (Hymenoptera, Halictidae) are model organisms for the evolution of altruism, reproductive castes, and eusocial colony organization. Halictine social behavior is not only extremely variable, but also ecologically and evolutionarily labile. Among social species, colony social organization ranges from communal societies of egalitarian females to eusocial and semisocial ones with reproductive queens and more or less sterile workers. A striking aspect of halictine social variation is the mutual exclusivity of communal and eusocial types of colony social organization within the same species, these two types of social behavior being characteristic of different genera and subgenera. We report a recently discovered exception to this rule in a population of Halictus sexcinctus (Fabricius) at Daimonia-Pyla in southern Greece, that contained both communal and eusocial colonies. Moreover, communal and eusocial females exhibit morphological differences that imply a preimaginal developmental switch, which could also underlie the two types of social behavior. That the communal and eusocial forms are not merely cryptic sister species with different social behavior is indicated by the comparison of mitochondrial DNA sequences of two sections of cytochrome oxidase I, which indicate that Greek specimens of both social types are more similar than they are to conspecifics from elsewhere in Europe. The phylogenetic position of Halictus sexcinctus suggests that this unusual communal/eusocial polymorphism may represent an unstable intermediate step in an evolutionary reversal from eusocial to solitary behavior. PMID- 12777630 TI - Transcriptional profiling of the Sonic hedgehog response: a critical role for N myc in proliferation of neuronal precursors. AB - Cerebellar granule cells are the most abundant neurons in the brain, and granule cell precursors (GCPs) are a common target of transformation in the pediatric brain tumor medulloblastoma. Proliferation of GCPs is regulated by the secreted signaling molecule Sonic hedgehog (Shh), but the mechanisms by which Shh controls proliferation of GCPs remain inadequately understood. We used DNA microarrays to identify targets of Shh in these cells and found that Shh activates a program of transcription that promotes cell cycle entry and DNA replication. Among the genes most robustly induced by Shh are cyclin D1 and N-myc. N-myc transcription is induced in the presence of the protein synthesis inhibitor cycloheximide, so it appears to be a direct target of Shh. Retroviral transduction of N-myc into GCPs induces expression of cyclin D1, E2F1, and E2F2, and promotes proliferation. Moreover, dominant-negative N-myc substantially reduces Shh-induced proliferation, indicating that N-myc is required for the Shh response. Finally, cyclin D1 and N-myc are overexpressed in murine medulloblastoma. These findings suggest that cyclin D1 and N-myc are important mediators of Shh-induced proliferation and tumorigenesis. PMID- 12777631 TI - Viral assembly of oriented quantum dot nanowires. AB - The highly organized structure of M13 bacteriophage was used as an evolved biological template for the nucleation and orientation of semiconductor nanowires. To create this organized template, peptides were selected by using a pIII phage display library for their ability to nucleate ZnS or CdS nanocrystals. The successful peptides were expressed as pVIII fusion proteins into the crystalline capsid of the virus. The engineered viruses were exposed to semiconductor precursor solutions, and the resultant nanocrystals that were templated along the viruses to form nanowires were extensively characterized by using high-resolution analytical electron microscopy and photoluminescence. ZnS nanocrystals were well crystallized on the viral capsid in a hexagonal wurtzite or a cubic zinc blende structure, depending on the peptide expressed on the viral capsid. Electron diffraction patterns showed single-crystal type behavior from a polynanocrystalline area of the nanowire formed, suggesting that the nanocrystals on the virus were preferentially oriented with their [001] perpendicular to the viral surface. Peptides that specifically directed CdS nanocrystal growth were also engineered into the viral capsid to create wurtzite CdS virus-based nanowires. Lastly, heterostructured nucleation was achieved with a dual-peptide virus engineered to express two distinct peptides within the same viral capsid. This work represents a genetically controlled biological synthesis route to a semiconductor nanoscale heterostructure. PMID- 12777632 TI - The economic value of ecological stability. AB - Seemingly intangible ecosystem characteristics that preoccupy ecologists, like ecosystem stability and the responsiveness of populations to environmental variation, have quantifiable economic values. We show how to derive these values, and how their consideration should change environmental decision making. To illustrate these concepts, we use a simple reserve design model. When resource managers choose a particular landscape configuration, their decision affects both the mean abundance of species and the temporal variation in abundances. Population stability and related phenomena have economic value, because management actions affect the variance of ecosystem components. In our example, a larger reserve size is recommended when accounting for the stability of the managed ecosystem. PMID- 12777633 TI - A role for Z-DNA binding in vaccinia virus pathogenesis. AB - The N-terminal domain of the E3L protein of vaccinia virus has sequence similarity to a family of Z-DNA binding proteins of defined three-dimensional structure and it is necessary for pathogenicity in mice. When other Z-DNA-binding domains are substituted for the similar E3L domain, the virus retains its lethality after intracranial inoculation. Mutations decreasing Z-DNA binding in the chimera correlate with decreases in viral pathogenicity, as do analogous mutations in wild-type E3L. A chimeric virus incorporating a related protein that does not bind Z-DNA is not pathogenic, but a mutation that creates Z-DNA binding makes a lethal virus. The ability to bind the Z conformation is thus essential to E3L activity. This finding may allow the design of a class of antiviral agents, including agents against variola (smallpox), which has an almost identical E3L. PMID- 12777634 TI - Carotid and femoral arterial wall mechanics in scleroderma. AB - OBJECTIVE: Large-vessel arterial disease is increasingly recognized as a major cause of morbidity in autoimmune rheumatic disorders. Recent evidence suggests that scleroderma (systemic sclerosis, SSc) may be linked to altered fibrillin-1 metabolism associated with a defect in chromosome 15q. If this is the case, we may expect to see changes in the arterial wall mechanics of large vessels not clinically involved in the disease process. We undertook a study to determine whether the biomechanical properties and intima-media thickness (IMT) of the elastic carotid artery and the muscular femoral artery are altered in subjects with limited (lcSSc) and diffuse (dcSSc) cutaneous SSc. METHODS: Measurements of carotid and femoral wall mechanics were made in 33 patients with lcSSc, 19 patients with dcSSc and 21 control subjects, using a duplex scanner coupled to a Wall Track system. Their age, gender, body mass index, heart rate, systolic and diastolic blood pressures, presumed cardiovascular load, and plasma creatinine, fasting cholesterol, triglyceride and glucose concentrations were also measured. RESULTS: There was a progressive and significant reduction (P < 0.001) in the elastic properties of the carotid artery from the control group (compliance, 16.24 +/- 4.39 %mmHg(-1) x 10(-2)) to the lcSSc group (10.89 +/- 2.43 %mmHg(-1) x 10(-2)) to the dcSSc group (7.65 +/- 2.08 %mmHg(-1) x 10(-2)), even after adjustment for the systemic physiological and biochemical variables studied, which are known to influence the mechanics of arterial walls. There was no apparent difference between the groups in the mean elastic indices of the femoral artery and the IMT of the carotid and femoral arteries. CONCLUSION: The elastic properties of the carotid artery are significantly altered in SSc, and the two major subsets of SSc may be distinguished by their carotid artery biomechanics. This suggests that connective tissue abnormality occurs at sites not previously assessed. PMID- 12777635 TI - Advanced glycation end-products pentosidine and N epsilon-carboxymethyllysine are elevated in serum of patients with osteoporosis. AB - OBJECTIVE: To investigate serum levels of the advanced glycation end-products (AGEs) pentosidine and N epsilon-carboxymethyllysine (CML) in patients classified into different osteoporosis subgroups according to histomorphometric data. METHOD: Serum samples were obtained from 116 osteoporotic patients (34 men, 82 women) classified by bone histomorphometry into subgroups with high turnover (HTO, n = 32), low turnover (LTO, n = 39), normal turnover (NTO, n = 9) and cellular uncoupled osteoporosis (CUO, n = 36). Pentosidine was measured by high performance liquid chromatography, and CML by a competitive enzyme-linked immunoassay. RESULTS: The entire osteoporosis group had significantly higher pentosidine and CML serum concentrations than healthy subjects. In contrast to healthy subjects, no correlation between levels of AGEs and age could be found. In subgroups characterized by increased bone resorption (HTO, CUO), serum pentosidine correlated significantly with the histomorphometric marker reflecting osteoclast activity/bone resorption (eroded surface as a percentage of trabecular surface). Moreover, in CUO a strong correlation between pentosidine and the mineral apposition rate was found. Surprisingly, in HTO the levels of CML and percentage of eroded surface were significantly negatively correlated. CONCLUSION: AGE-modified proteins may be a cause of disturbed bone remodelling in osteoporosis. Our findings do not support the alternative hypothesis that increased AGEs in serum indicate only a more intensive releasing of AGEs in circumstances of increased bone resorption. PMID- 12777636 TI - Inhibition of cytokine production by methotrexate. Studies in healthy volunteers and patients with rheumatoid arthritis. AB - OBJECTIVES: To analyse whether the beneficial effects of methotrexate in rheumatoid arthritis (RA) could be due to inhibition of inflammatory cytokine production. METHODS: Cytokine production was studied using whole blood (WB) and mononuclear cells (MNC) of healthy volunteers and RA patients. Cultures were stimulated with either bacterial products such as lipo-oligosaccharide (LOS) or Staphylococcus aureus Cowan I (SAC) to activate monocytes or with monoclonal antibodies to CD3 and CD28 to induce polyclonal T-cell activation. We analysed the effect of methotrexate on cytokine production in these systems. RESULTS: We showed that methotrexate inhibits production of cytokines induced by T-cell activation. Among the cytokines inhibited were interleukin 4 (IL-4), IL-13, IFN gamma, tumour necrosis factor-alpha (TNF alpha) and granulocyte-macrophage colony stimulating factor. Inhibition was seen at concentrations easily achieved in plasma of RA patients taking the drug. IL-8 production was hardly influenced by methotrexate. Furthermore, inhibition was dependent on the stimulus; IL-6, IL-8, IL-1 beta and TNF alpha production induced by LOS or SAC was only slightly decreased by methotrexate. The addition of folinic acid or thymidine and hypoxanthine reversed the inhibitory effects of methotrexate on cytokine production. Concentrations of methotrexate required for inhibition varied between donors. Oral intake of 10 mg methotrexate by RA patients led to marked inhibition of cytokine production in blood drawn after 2 h. CONCLUSIONS: Methotrexate turns out to be an efficient inhibitor of cytokine production induced by T-cell activation in freshly drawn blood. This is due to inhibition of the de novo synthesis of purines and pyrimidines. Cytokines produced by monocytes are hardly affected by methotrexate. PMID- 12777637 TI - Prognosis and outcome of 26 patients with systemic necrotizing vasculitis admitted to the intensive care unit. AB - OBJECTIVES: To investigate presenting features, prognostic factors and outcomes of patients with systemic necrotizing vasculitis (SNV) admitted to the intensive care unit (ICU). METHODS: We retrospectively reviewed the medical records of all 210 SNV patients followed in our university hospital and admitted to the ICU between 1982 and 2001, with respect to clinical features, ICU disease severity scores (APACHE II and SAPS II), the Birmingham vasculitis activity score (BVAS), the five-factors score (FFS) and outcomes. RESULTS: Twenty-six patients (16 men, 10 women) with a mean age of 46.3+/-16.5 yr were included. The reasons for ICU admission were: active SNV, 20 (77%); infection, 3 (12%); others, 3 (12%). SNV was diagnosed in 11 (42%) patients in the ICU. The mean APACHE II and SAPS II scores were significantly higher for patients who died in the ICU (P = 0.01 and P = 0.01 respectively). After a mean follow-up of 31.4+/-29.2 months, the overall mortality rate was 39% (10 patients). Among patients admitted to the ICU with active SNV, BVAS calculated at ICU admission was significantly higher for non survivors at the end of follow-up (26.9+/-13.0 vs 14.7+/-4.6, P = 0.02). CONCLUSION: The main reason for admitting SNV patients to the ICU was active vasculitis, which was often the first manifestation of SNV and led to its diagnosis. ICU disease severity scores at admission were associated with mortality in the ICU but did not predict long-term outcome, unlike BVAS, which accurately predicted long-term outcome but not ICU prognosis for patients admitted to the ICU with active SNV. PMID- 12777638 TI - Evaluation of technetium-99m-ciprofloxacin (Infecton) for detecting sites of inflammation in arthritis. AB - OBJECTIVE: To study the frequency of technetium-99m-positive ciprofloxacin scans (Infecton scintigraphy) thought to be specific for bacterial DNA in patients with arthritis and to assess the clinical relevance of positive scans. METHODS: Four groups of adults with arthritis were studied. Group 1: 53 patients with inflammatory arthritis, 36 with spondylarthropathy (SpA) and 17 with rheumatoid arthritis (RA); group 2: five patients with crystal arthropathy; group 3: those patients with osteoarthritis (OA) of the knee, wrist or spine; and group 4: 28 patients who had no arthritis but were being investigated for renal infection. Patients were injected with 10 mCi 99Tcm-ciprofloxacin with isotope uptake analysis at 4 h. Clinically swollen joints were assessed by a rheumatologist and the positive scans assessed by a physician in nuclear medicine. RESULTS: Increased Infecton uptake was noted in inflamed joints independent of the pathology. It was seen in 10 of 17 patients with SpA, 12 of 17 with RA, all five with crystal arthropathy, eight with knee OA, two with wrist OA, none with spinal OA and none in uninflamed joints. A close correlation between clinically swollen joints and articular Infecton uptake was noted (P = 0.0003), with the uptake being in the distribution of the synovial perimeter. Additional uptake was noted in the abdomen (n = 9) and pulmonary region (n = 2) of SpA patients. CONCLUSION: The Infecton scan is not specific for infection but may be a reliable procedure for identifying the presence and distribution of the inflammation within joints. It has the potential for monitoring the response of inflamed joints to treatment. PMID- 12777639 TI - SACRAH: a score for assessment and quantification of chronic rheumatic affections of the hands. AB - OBJECTIVES: To establish a questionnaire to quantify the extent of the function and activities of the hand in patients with degenerative or inflammatory disease of the hand and finger joints. METHODS: One hundred and seventy-two patients with osteoarthritis (OA, n = 69) or rheumatoid arthritis (RA, n = 103) completed a new questionnaire, the SACRAH, that included 23 visual analogue scales covering the extent of hand function, stiffness and level of pain. SACRAH scores may range from 0 to 100. RESULTS: Comparing all studied patients, there was no significant difference in SACRAH scores between OA and RA patients (34 vs 32, not significant). Scores for both patient groups differed significantly from those for 30 healthy controls. Among patients taking NSAIDs only, individuals suffering from OA (n = 50) scored significantly lower than RA patients (n = 42) (36 vs 48, P < 0.004). Sixty-one RA patients taking DMARDs scored lower than the RA patient group treated with NSAIDs only (20 vs 48, P < 0.0001). Thirty-two RA patients were evaluated longitudinally at their first visit and 3 months after the initiation of DMARDs. Following therapy, SACRAH scores were significantly reduced from 50 to 11 (P < 0.0001). CONCLUSIONS: The questionnaire enables the quantification of compromised hand function, stiffness and pain in OA and RA patients, and is sensitive to therapy-related changes in RA patients. PMID- 12777640 TI - Pregnancy outcome in adult-onset idiopathic inflammatory myopathy. AB - OBJECTIVES: To assess the prevalence of pregnancy in 28 females with dermatomyositis (DM) and polymyositis (PM), assess the outcome in those who became pregnant after the onset of the disease and review the literature of all published cases. METHOD: Fifty-four patients with myositis have been under long term follow-up from 1976-2001 (28 female, nine male). RESULTS: Twenty-eight female patients were divided into 15 with pure DM/PM (seven PM, eight DM) and 13 with an overlap syndrome. The majority of patients had the onset of the disease after childbearing years (mean age of 32 yr for the overlap group and 41 for the DM/PM group). Only four of our patients (14.3%) have been pregnant after the onset of the disease. One patient had a spontaneous abortion, but was on methotrexate and had active disease; one had a late pregnancy loss, but had active disease; and the other two had uneventful pregnancies during a time when the disease was in remission. CONCLUSION: Fetal prognosis in the main reflects the level of maternal disease. The more active the myositis during the pregnancy, the greater the chance of fetal loss. PMID- 12777641 TI - Paucity of Sjogren-like syndrome in a cohort of HIV-1-positive patients in the HAART era. Part II. AB - OBJECTIVE: This study was performed in order to investigate the prevalence of Sjogren-like syndrome (SLS) in the highly active anti-retroviral therapy (HAART) era in a cohort of HIV-1-positive Greek patients. METHODS: One hundred and thirty one unselected patients were screened by the validated European Union (EU) criteria for Sjogren's syndrome. Of the 31 who gave a positive EU-validated questionnaire, 17 consented to undergo minor salivary gland biopsy and other tests. RESULTS: Only two patients had a positive salivary gland biopsy and both belonged to the non-compliant HAART group, whereas none of the compliant HAART patients had histological findings. CONCLUSIONS: It is concluded that SLS, the prevalence of which in the pre-HAART era was 7.8%, has disappeared, possibly as a result of the protective action of HAART. PMID- 12777643 TI - What is pain management, and what is its relevance to the rheumatologist? PMID- 12777642 TI - Protein interaction for an interferon-inducible systemic lupus associated gene, IFIT1. AB - OBJECTIVE: To identify disease-related genes and immune-regulatory pathways in the pathogenesis of systemic lupus erythematosus (SLE) by using gene expression profiling and protein-protein interaction analysis. METHODS: Peripheral white blood cell gene expression profiles of 10 SLE patients were determined by oligonucleotide microarray analysis. Clustering of the gene expression profile was compared with the clinical immune phenotype. SLE-induced genes that were over or under-expressed were determined and independently validated using a real-time polymerase chain reaction (PCR) method. To study their potential function and the possible pathways involved, a candidate gene was cloned and a GST (glutathione S transferase) fusion protein was expressed in Escherichia coli. The fusion protein was further purified using the glutathione Sepharose 4B system, and was treated as bait to capture prey from SLE peripheral white blood cell lysate. MALDI-TOF (matrix-assisted laser desorption/ionization-time-of-flight) mass spectrometry was then performed to determine the prey protein. RESULTS: Similarity was found between the gene expression profile and the immune phenotype clusters of the SLE patients. More than 20 disease-associated genes were identified, some of which have not been related to SLE previously. Of these genes, a cluster of interferon induced genes were highly correlated. IFIT1 (interferon-induced with tetratricopeptide repeats 1) was one of these genes, and overexpression of its mRNA was confirmed independently by real-time PCR in a larger population (40 SLE patients and 29 normal controls). An IFIT1 protein- protein interaction study showed that IFIT1 may interact with Rho/Rac guanine nucleotide exchange factor. CONCLUSION: The gene expression profile seems to be the molecular basis of the diverse immune phenotype of SLE. On the basis of the SLE-related genes found in this study, we suggest that the interferon-related immune pathway is important in the pathogenesis of SLE. IFIT1 is the first gene described as a candidate gene for SLE, and may function by activating Rho proteins through interaction with Rho/Rac guanine nucleotide exchange factor. IFIT1 and the interferon-related pathway may provide potential targets for novel interventions in the treatment of SLE. PMID- 12777644 TI - Acupuncture to reduce nausea during chemotherapy treatment of rheumatic diseases. AB - OBJECTIVE: To study if acupuncture, combined with ondansetron treatment, reduces nausea and vomiting associated with cyclophosphamide infusion in patients with rheumatic diseases. METHODS: Thirty-nine patients were treated with acupuncture at point PC 6 and/or in the ear to decrease nausea and vomiting. The patients reported the severity of nausea and number of bouts of vomiting at the start of chemotherapy and after 4, 8, 24, 48 and 72 h. RESULTS: Compared with ondansetron treatment alone, the combined acupuncture-ondansetron treatment significantly decreased both the severity of nausea and the number of bouts of vomiting 24 and 48 h after the subjects had received acupuncture at the first treatment session (nausea: P < 0.0001; vomiting: P < 0.0035). Nearly the same results were seen 48 and 72 h after the subjects had had their last treatment of acupuncture (nausea P < 0.0080). Similar results were found after 24 to 48 h, when a comparison was made between two sessions close in time (nausea: P < 0.0001 after 24 h, P < 0.0003 after 48 h; vomiting: P < 0.0007). CONCLUSIONS: Our results clearly indicate that combined treatment with acupuncture and ondansetron reduces the severity and the duration of chemotherapy-induced nausea as well as the number of bouts of vomiting as compared with ondansetron therapy alone, in patients with rheumatic diseases. PMID- 12777645 TI - The safety of massage therapy. AB - OBJECTIVES: After many years out of the limelight, massage therapy is now experiencing a revival. The aim of this systematic review is to evaluate its potential for harm. METHODS: Computerized literature searches were carried out in four databases. All articles reporting adverse effects of any type of massage therapy were retrieved. Adverse effects relating to massage oil or ice were excluded. No language restrictions were applied. Data were extracted and evaluated according to predefined criteria. RESULTS: Sixteen case reports of adverse effects and four case series were found. The majority of adverse effects were associated with exotic types of manual massage or massage delivered by laymen, while massage therapists were rarely implicated. The reported adverse events include cerebrovascular accidents, displacement of a ureteral stent, embolization of a kidney, haematoma, leg ulcers, nerve damage, posterior interosseous syndrome, pseudoaneurism, pulmonary embolism, ruptured uterus, strangulation of neck, thyrotoxicosis and various pain syndromes. In the majority of these instances, there can be little doubt about a cause-effect relationship. Serious adverse effects were associated mostly with massage techniques other than 'Swedish' massage. CONCLUSION: Massage is not entirely risk free. However, serious adverse events are probably true rarities. PMID- 12777646 TI - Intra-articular corticosteroid preparations: different characteristics and their effect during inflammation induced by monosodium urate crystals in the rat subcutaneous air pouch. AB - OBJECTIVE: To examine the effects of three commonly used intra-articular depot corticosteroid preparations tested in a rat air pouch model and their effect against monosodium urate (MSU) crystal-induced inflammation. Rheumatologists use intra-articular corticosteroid preparations to relieve pain and inflammation of acute monoarthritis without really knowing their effects on the synovial fluid and membrane or the differences between distinct preparations. This work compares the effect of three commonly used corticosteroid preparations in vivo, showing that they behave differently. METHODS: A subcutaneous air pouch was formed in male Sprague-Dawley rats. A first group of 6-day-old air pouches were injected with 10 ml of 6 mg/ml normal saline solution, 6 mg/ml betamethasone containing both depot betamethasone acetate and soluble betamethasone phosphate (Celestone) in 9 ml of normal saline solution, 20 mg/ml of prednisolone tebutate (Hydeltra) in 9 ml of normal saline solution or 20 mg/ml of triamcinolone hexacetonide (Aristospan) in 9 ml of normal saline solution. A second group (group 2) of air pouches were injected with 15 mg of synthetic MSU crystals and 24 h later they were reinjected with 1 ml of the same three corticosteroid suspensions. For each condition four rats were killed at 6, 24, 48 h and 7 days. Pouch fluid and tissue were analysed. RESULTS: In the first 6 h after normal saline solution or corticosteroid injection into the air pouch there were mildly increased leucocyte counts in the air pouch fluid. Betamethasone-injected pouches showed no cells in the fluid after 6 h and no crystals after 24 h, triamcinolone-injected pouches still showed rare cells at 7 days. Both triamcinolone and prednisolone crystals persisted in higher numbers and lasted longer in the fluid than did betamethasone (P<0.05). In group 2 MSU crystal phagocytosis in the fluid was decreased in the betamethasone- (P<0.01), prednisolone- (P<0.003) and triamcinolone- (P<0.006) injected pouches when compared with the MSU crystal-injected pouches alone. Pouches injected with MSU crystals alone showed the most intense tissue inflammation at all times. After MSU, betamethasone-injected pouches had a rapid but mild decrease in the number of lining cells and inflammation. In contrast, triamcinolone- and prednisolone-injected pouches showed a very thin tissue with few or no vessels and almost no inflammation at 7 days. The pouches injected with MSU crystals and any of the corticoid preparations had three times more tophus like structures and persistent crystals identified than the ones injected with MSU crystals alone. CONCLUSION: Each of the corticosteroid preparations by themselves produced very mild transient inflammation. The betamethasone preparation with a soluble steroid component had a quicker but milder anti inflammatory effect on MSU crystal-induced inflammation. In contrast to the doses used, prednisolone tebutate and triamcinolone hexacetonide preparations dramatically suppressed urate crystal-induced inflammation at 7 days, but both produced atrophy and necrosis of the membrane, yielding a very thin membrane with almost no vessels. When used for MSU crystal-induced inflammation these corticosteroid preparations suppressed some aspects of inflammation but may actually promote the persistence of MSU crystals and the formation of tophi. PMID- 12777647 TI - Osmic acid revisited: factors that predict a favourable response. AB - OBJECTIVE: Chemical synovectomy with osmic acid has been used for many years in the treatment of chronic synovitis that is unresponsive to systemic treatment and intra-articular corticosteroid. Our aims were to compare the safety and the efficacy of this procedure with that of alternative treatment modalities, and to identify any factors that may predict the response to treatment. METHODS: A retrospective review of 103 osmic acid injections was performed in 65 patients with chronic knee synovitis, between 1992 and 1998. After a clinical review 6 weeks after injection, the length of remission was determined by telephone survey. Thirty-six months of follow-up was available for 96 injections, with a minimum of 12 months for the remainder. Remission was defined as complete absence of pain and swelling. RESULTS: Sixty-nine (67.0%) knee joints remained completely free of pain and swelling at 6 months, falling to 52 (50.5%) at 12 months, 32 (31.1%) at 24 months, and 19 (18.4%) at 36 months. Knees with mild radiological changes experienced significantly better results compared with those with moderate or severe changes (P=0.006 and 0.046 respectively). In patients undergoing bilateral injections, there was a correlation between the duration of remission achieved for each of the two knees (r=0.83, P < 0.01). A correlation was also observed between the responses obtained after first and repeated synovectomies of the same knee (r=0.62, P < 0.05). Eighty-six per cent of injections were uncomplicated. Side-effects included pain, which followed 13 injections (12.4%), and two skin burns. CONCLUSIONS: Long-term complete remission was achieved in 18.4% of knees. This treatment may be best reserved for joints with less severe radiological changes. The response to a first osmic acid injection has a strong predictive value when considering further treatment of the same or the contralateral joint. PMID- 12777648 TI - Towards better management of chronic atrial fibrillation. PMID- 12777649 TI - Prevalence of impacted teeth and associated pathologies--a radiographic study of the Hong Kong Chinese population. AB - OBJECTIVES: To investigate the prevalence and pattern of impacted teeth and associated pathologies in the Hong Kong Chinese population. SETTING: The Reception and Primary Care Clinic, Prince Philip Dental Hospital, Hong Kong. DESIGN: Retrospective study. SUBJECTS AND METHODS: The records of 7486 patients were examined to determine whether the chief complaints were related to impacted teeth and associated pathologies, which were investigated using panoramic radiographs. RESULTS: A total of 2115 (28.3%) patients presented with at least one impacted tooth. Among the 3853 impacted teeth, mandibular third molars were the most common (82.5%), followed by maxillary third molars (15.6%), and maxillary canines (0.8%). Approximately 8% of mandibular second molars associated with impacted third molars had periodontal bone loss of more than 5 mm on their distal surfaces. Caries were also found on the same surfaces in approximately 7% of the second molars. Approximately 30% of patients with dental impaction had symptoms, and 75% had complaints limited to one side of the mouth. CONCLUSIONS: The prevalence of impacted teeth was high, and there was a predilection for impacted third molars in the mandible. More than 50% of maxillary third molars had erupted, creating potential trauma of the pericoronal tissues of the partially erupted mandibular third molars. Caries and periodontal diseases were commonly seen in relation to the impacted third molars, whereas cystic pathology and root resorption were rarely observed. PMID- 12777650 TI - Outcome for Hong Kong residents undergoing cadaveric liver transplantation in mainland China. AB - OBJECTIVE: To review the outcome for Hong Kong residents undergoing cadaveric liver transplantation in mainland China. DESIGN: Retrospective study. SETTING: Liver Transplant Centre, university teaching hospital, Hong Kong. SUBJECTS AND METHODS: A retrospective review of medical records was undertaken for patients at Queen Mary Hospital who underwent cadaveric liver transplantation in China between 1 January 1997 and 31 December 2001. RESULTS: Fifteen patients from Queen Mary Hospital underwent cadaveric liver transplantation in China during the study period. Eleven were men and four were women. Their mean age was 51 years. Disease indications included hepatitis B-related liver cirrhosis (n=7), hepatitis B related liver cirrhosis with hepatocellular carcinoma (n=5), hepatitis C-related liver cirrhosis (n=1), hepatitis C-related liver cirrhosis with hepatocellular carcinoma (n=1), and polycystic liver and kidney disease (n=1). Nine patients were already waiting for liver transplantation at Queen Mary Hospital, and two of the nine patients were on the 'urgent' list. The overall survival rate was 80.0% at 6 months and 73.3% at 12 months. There were four (27%) deaths, two of which occurred in China. Of the 11 surviving patients, nine (82%) developed complications. Nineteen complications were seen in the 13 patients who were managed in Hong Kong following their return from China. Infective and biliary complications accounted for 58% and 26% of complications, respectively. Major complications necessitated prolonged hospitalisation for four patients and two required further laparotomy. CONCLUSION: Although cadaveric liver transplantation in China is an option for Hong Kong residents, patients and clinicians should be aware of the possible outcomes and resource implications. PMID- 12777651 TI - The value of flexible sigmoidoscopy for patients with bright red rectal bleeding. AB - OBJECTIVE: To review the diagnostic yield of flexible sigmoidoscopy in patients presenting with bright red rectal bleeding. DESIGN: Retrospective study. SETTING: University teaching hospital, Hong Kong. SUBJECTS AND METHODS: Patients who underwent flexible sigmoidoscopy between January 1995 and April 1996 for investigation of bright red rectal bleeding were recruited. The extent of the endoscopic examination, complications, and endoscopic findings were recorded. RESULTS: A total of 1052 patients were included in the study. The mean length of endoscopic examination was 55 cm. There were no complications attributed to the procedure. Thirteen (1.2%) patients aged from 41 to 87 years were found to have malignant tumours that were not palpable on digital examination. All the tumours were moderately differentiated adenocarcinoma. Two patients had synchronous liver metastasis at presentation. Adenomatous polyps were detected in 81 (7.7%) patients, of whom 76 were older than 40 years. The majority of polyps were tubular adenomas associated with mild or moderate dysplasia. Other endoscopic findings included hyperplastic and juvenile polyps, proctocolitis, diverticulosis, irradiation colitis, ischaemic colitis, rectal ulcers, and infective colitis. The overall diagnostic yield was 21.1%. No mucosal lesion was detected by flexible sigmoidoscopy in 78.9% of patients in whom the rectal bleeding was due to either haemorrhoids or anal fissure. CONCLUSIONS: Cancer was detected in 1.2% and adenomatous polyps in 7.7% of patients with bright red rectal bleeding using flexible sigmoidoscopy. All cancers and 94% of adenomatous polyps were detected in patients older than 40 years. Flexible sigmoidoscopy appears to be a valuable initial investigation for bright red rectal bleeding in patients older than 40 years. PMID- 12777652 TI - Ureteroscopy as an out-patient procedure: the Singapore General Hospital Urology Centre experience. AB - OBJECTIVE: To study the safety and efficacy of ureteroscopy as an out-patient procedure. DESIGN: Retrospective study. SETTING: Teaching hospital, Singapore. SUBJECTS AND METHODS: Operating facilities for out-patient ureteroscopy were introduced in late 1998. A retrospective review of all patients undergoing ureteroscopy from March 1999 to December 2000 was conducted. Clinical records were reviewed for operating time, surgical outcome and complications, and length of hospital stay and re-admissions. RESULTS: Two hundred and ten patients underwent ureteroscopy for a variety of indications--115 patients were admitted to hospital as in-patients and 95 patients were treated as out-patients. Ureteroscopy and laser lithotripsy were performed using general or regional anaesthesia. Patients assessed to be American Society of Anesthesiology class I or II were eligible for an out-patient procedure. American Society of Anesthesiology class was found to be lower for those treated as out-patients compared with those undergoing in-patient procedures--63% versus 28% for class I and 36% versus 69% for class II. Mean operating time was 37 minutes for out patient procedures and 57 minutes for in-patient procedures (P<0.01). Complications were low for out-patients, with six unplanned re-admissions, mostly for minor complications. All patients were discharged the following day. CONCLUSION: Ureteroscopy can be offered selectively as an out-patient procedure to patients with low surgical risk, especially American Society of Anesthesiology class I patients, and others expected to have an uncomplicated surgical procedure. PMID- 12777653 TI - Antithrombotic treatment of atrial fibrillation in a regional hospital in Hong Kong. AB - OBJECTIVE: To measure the use, appropriateness, and safety of antithrombotic therapy in Hong Kong Chinese patients with atrial fibrillation. DESIGN: Retrospective review. SETTING: Regional hospital, Hong Kong. SUBJECTS AND METHODS: Medical records of all patients with atrial fibrillation admitted to acute internal medicine wards in April 2000 and between July and October 2001 were reviewed for details of antithrombotics given, results of international normalised ratio monitoring for patients receiving warfarin, side-effects, and additional risk factors for complications of atrial fibrillation. Statistical analysis was undertaken to assess factors predictive of antithrombotic use. RESULTS: A total of 207 patients with chronic atrial fibrillation were included in the study. Of these, 44.0% of patients with non-valvular atrial fibrillation without contra-indications for warfarin use were receiving warfarin, 34.1% were receiving aspirin, and 22.0% were receiving no antithrombotic therapy. The majority of patients (69.1%) were treated appropriately according to the American College of Chest Physicians guidelines. The major side-effect rates for warfarin and aspirin were 2.14% and 1.72% per patient-year, respectively, which were comparable with western studies of usual clinical practice. The ischaemic stroke rate for patients taking warfarin or aspirin were 1.40% and 6.02% per patient year, respectively. The median international normalised ratio was 1.96. The median frequency of international normalised ratio measurement was 45.58 days. CONCLUSIONS: This study found that antithrombotic use in a Hong Kong regional hospital for patients with atrial fibrillation was similar to that reported from western institutions. Complication and stroke rates were also comparable to the western data relating to usual clinical practice. PMID- 12777654 TI - Hong Kong Chinese teachers' attitudes towards life-sustaining treatment in the dying patients. AB - OBJECTIVE: To evaluate the attitudes of Hong Kong Chinese teachers towards life sustaining treatment in the dying patients. DESIGN: Prospective structured questionnaire survey. SETTING: Hong Kong Institute of Education. SUBJECTS AND METHODS: All teaching staff at the Hong Kong Institute of Education were sent the survey questionnaire. The questionnaire gathered demographic data, information on experience of 'life and death' decision-making, and views on life-sustaining treatment decisions. Respondents were also requested to respond to statements on life-sustaining treatment using a 5-point Likert Scale (1 representing strong disagreement and 5 representing strong agreement). RESULTS: A total of 436 questionnaires were sent to teaching staff at the Hong Kong Institute of Education. The response rate was 27.8%. More than half (65.8%) of the respondents were aged between 30 and 49 years. There was an equal proportion of men and women among respondents. The respondents agreed with statements supporting end-of-life decisions (mean aggregate score, 3.13; standard deviation, 1.24; P<0.0001), and disagreed with statements against such decisions (mean aggregate score, 2.81; standard deviation, 1.22; P<0.0001). If the patient is competent, half of the respondents agreed that the patient alone should make the decision, while 27.2% believed that it should be a joint decision made by the patient, the family, and the doctor. Conversely, if the patient is incompetent, 52.6% agreed that it should be a joint decision made by the family and the doctor. There was strong support for advanced directives, whereby decisions in relation to life-sustaining treatment were legally recorded in advance (mean score, 3.62; P=0.0001). CONCLUSION: The teachers in this survey appear to support the practice of withdrawing and withholding life-sustaining treatment in the dying patients when medical treatment is considered futile. Although patient autonomy in decision making was supported by the greatest number of respondents, joint decision-making by the patient, family members, and doctors was also favoured by a substantial group. There was strong support for the use of advanced directives with respect to life-sustaining treatment. PMID- 12777655 TI - Ethical issues related to the use of placebo in clinical trials. AB - Controversy exists regarding the ethics of using placebo control groups in clinical trials when effective treatments exist. The debate was fuelled by the announcement of the fifth revision of the Declaration of Helsinki in 2000. This study reviews the history and scientific background surrounding the controversy and investigates the prevailing attitudes of Hong Kong researchers regarding this issue. The controversy has centred on several issues. The first involves the methodological superiority of placebo-controlled trials in discerning treatment effects. Secondly, it is unclear whether the treatment effects encompass absolute treatment effects (including placebo effects) or are confined to treatment specific effects (excluding placebo effects). Thirdly, there are concerns that subjects in the placebo group could be exposed to a high risk for developing serious adverse events. Fourthly, it is unclear whether the standard of best available treatment should be local or international. Preliminary research findings suggest that the opinions of Hong Kong researchers are divided on the use of placebo control groups in clinical trials when effective treatment exists. Further research on the topic is therefore warranted, and training and consensus meeting may be necessary to minimise the confusion related to this issue. PMID- 12777656 TI - Treatment of severe acute respiratory syndrome with convalescent plasma. AB - In March 2003, an outbreak of severe acute respiratory syndrome started in Hong Kong. A 57-year-old woman had a typical presentation, including fever, non productive cough, malaise, lymphopenia, and raised liver aminotransferases. The clinical course and successful treatment with convalescent plasma, ribavirin, and corticosteroids are discussed. PMID- 12777657 TI - Severe acute respiratory syndrome in a doctor working at the Prince of Wales Hospital. AB - Severe acute respiratory syndrome is a new disease that is highly contagious and is spreading in the local community and worldwide. This report is of a hospital medical officer with severe acute respiratory syndrome. He presented with sudden onset of fever, chills, myalgia, headache, and dizziness in early March 2003. He developed progressive respiratory symptoms and bilateral pulmonary infiltrates during the second week of his illness. Blood tests showed lymphopenia, mild thrombocytopenia, and prolonged activated partial thromboplastin time with normal d-dimer level. His chest condition gradually responded to ribavirin and corticosteroids, and serial chest X-ray showed resolving pulmonary infiltrates. The importance of early diagnosis lies in the potential for early treatment, leading to better response. PMID- 12777658 TI - Sitosterolaemia and xanthomatosis in a child. AB - A 4-year-old boy presented with multiple tuberous xanthomata and a fasting plasma sterol concentration of 18.3 mmol/L, consisting primarily of cholesterol. Two months after changing from an unrestricted diet to a cholesterol-lowering diet, the plasma sterol concentration decreased to 4 mmol/L. Fasting plasma cholesterol levels for his father and mother were 7.3 mmol/L and 6.0 mmol/L, respectively. The degree and rapidity of the child's response to dietary control, together with the fasting cholesterol results of both parents suggested a diagnosis of sitosterolaemia. Gas chromatography and mass spectrometry of the patient's plasma sterol levels showed that the percentage of beta-sitosterol was raised at 12.76%, as was campesterol (6.26%), and stigmasterol (0.71%), confirming the diagnosis of sitosterolaemia. The addition of cholestyramine 4 g/day to a low sterol diet maintained the plasma sterol concentration at 4 to 5 mmol/L, and gradual regression of the xanthoma was observed. These findings indicate that a diagnosis of sitosterolaemia, a treatable cause of premature atherosclerosis, should be considered in children with severe hypercholesterolaemia whose plasma cholesterol level is highly responsive to dietary manipulation. PMID- 12777659 TI - Transient hypotension after high-speed head-on motor vehicle collision. AB - A 43-year-old woman was taken to the accident and emergency department after a high-speed motor vehicle collision. She had only minor external injuries, but developed an episode of hypotension, which rapidly responded to fluid infusion. Chest X-ray showed a widened superior mediastinum. Computed tomography confirmed thoracic aortic injury. In patients with high-speed deceleration injury, even transient hypotension should be taken seriously and a diligent search for the source of bleeding should be immediately undertaken. PMID- 12777660 TI - Unimuscular neuromuscular insult of the leg in partial anterior compartment syndrome in a patient with combined fractures. AB - A complicated case of ipsilateral fractures of the left femur and tibia after a road traffic accident is reported. The patient presented with numbness of the first web of his left foot and contracture of the extensor hallucis longus muscle, with fixed length deformity after intramedullary nailing of the femur and tibia. The extensor digitorum longus and tibialis anterior muscles were spared. Tinel's sign could be elicited at the mid-portion of the anterior compartment of the injured leg. This indicated that the distal half of the anterior tibial nerve (deep peroneal nerve), together with the extensor hallucis muscle of the anterior compartment of the leg, had been damaged. The subsequent management of this patient is described. PMID- 12777661 TI - Kennedy's disease. AB - Kennedy's disease is an X-linked, neurodegenerative disorder, characterised by lower motor neuron syndrome. This report gives the clinical details of six male patients with Kennedy's disease diagnosed at Princess Margaret Hospital. Three were initially diagnosed with other neurological diseases, with the diagnosis of Kennedy's disease made after genetic testing. This hereditary disease should be considered in male patients with muscle weakness, particularly those with a presentation suggesting atypical motor neuron disease. PMID- 12777662 TI - Ruptured phaeochromocytoma--a lesson in acute abdomen. AB - Phaeochromocytoma may present as acute abdomen. This report is of a patient with spontaneous rupture of phaeochromocytoma who presented with abdominal pain and a tender abdominal mass. Ruptured phaeochromocytoma is a rare surgical emergency, with only 30 cases reported in the literature. The classical clinical triad of signs is intense vasoconstriction, tachycardia, and labile blood pressure. Computed tomography scanning of the abdomen is the investigation of choice, and a high index of suspicion is the key to diagnosis. Prompt recognition, appropriate supportive measures, and early surgical intervention can improve the likelihood of survival. PMID- 12777663 TI - Combination therapy for hypertension. PMID- 12777664 TI - Personal risk management--with case histories. PMID- 12777665 TI - Inoue-balloon mitral valvuloplasty in double-orifice mitral stenosis. AB - We report 7 symptomatic patients with stenotic double-orifice mitral valve of incomplete bridge type. In each patient, the fibrous bridge tissue between the valve leaflets was successfully split using an Inoue balloon valvuloplasty technique with stepwise dilations applied only to the posteromedial orifice. PMID- 12777666 TI - Contrast-media induced nephropathy in patients undergoing coronary angiography. AB - Contrast-medium induced nephropathy (CIN) in patients undergoing cardiac catheterization has an extremely variable incidence, which is very elevated in high-risk subgroups. Despite its benign course in most cases, it is associated with a longer hospital stay, and elevated in-hospital and late mortality rates. This paper reviews the physiopathogenesis of CIN, as well as the clinical manifestations and preventive measures currently available. PMID- 12777667 TI - Prevention of radiocontrast-induced nephropathy with N-acetylcysteine in patients undergoing coronary angiography. AB - OBJECTIVES: Acetylcysteine in patients undergoing computerized tomography with intravenous contrast reduces the incidence of acute renal dysfunction. We examined the effect of N-acetylcysteine in patients undergoing coronary angiography. METHODS: Fifty-five consecutive patients receiving 3 doses of N acetylcysteine prior to cardiac catheterization were compared to 55 historical controls. All patients in both groups had baseline serum creatinine > 1.2 mg/dl and received intravenous hydration before and after the procedure. Serum creatinine levels at baseline and 48 hours after the procedure were compared. RESULTS: Univariate analysis of clinical variables revealed no significant differences between the groups except for a higher baseline creatinine in the treatment group (2.0 0.7 vs. 1.8 0.4 mg/dl; p = 0.04). There was no difference in the amount or type of contrast used. The mean change in creatinine after 48 hours was -0.4 0.3 versus +0.1 0.3 mg/dl for treatment and control groups (p < 0.001). In patients with baseline creatinine > 2 mg/dl, the benefit was larger (-0.4 0.4 vs. +0.5 0.3 mg/dl; p < 0.001). Multivariate analysis confirmed pre-treatment with N-acetylcysteine as an independent predictor of renal protection (p < 0.001). CONCLUSIONS: Prophylactic use of acetylcysteine prevented reduction of renal function after coronary angiography. The benefit was greater in patients with baseline serum creatinine > 2 mg/dl. PMID- 12777668 TI - Radiocontrast-induced nephropathy. PMID- 12777669 TI - Contrast-induced nephropathy: back to basics. PMID- 12777670 TI - Comparison of two platelet glycoprotein IIb/IIIa inhibitors, eptifibatide and abciximab: outcomes, complications and thrombocytopenia during percutaneous coronary intervention. AB - BACKGROUND: Platelet glycoprotein IIb/IIIa inhibitors have significantly reduced the incidence of 30-day ischemic events during percutaneous coronary interventions (PCI). However, each of the three currently available agents has different pharmacological characteristics, safety, efficacy and costs. There has not been a direct comparison between eptifibatide and abciximab in the rates of major adverse cardiac events, major complications and thrombocytopenia. METHODS: A total of 642 consecutive patients underwent PCI at our institution between January 2000 and December 2001 and were treated with either eptifibatide (n = 342) or abciximab (n = 300) during the procedure. The selection of the IIb/IIIa inhibitor was arbitrary and left to the discretion of the operator. Complete blood counts were performed by routine protocol on all patients 2 and 4 hours after initiation of the drug. We analyzed the in-hospital clinical outcomes and the incidence of thrombocytopenia in this cohort. RESULTS: Baseline clinical and angiographic characteristics and concomitant drug treatment were similar between the 2 groups, except for a higher incidence of diabetes in the eptifibatide group. The rates of in-hospital death (1.2% eptifibatide group versus 1.0% abciximab group; p = 0.7), stroke (0% for both groups), target vessel revascularization (1.2% eptifibatide group versus 1.0% abciximab group; p = 0.8) and major bleeding complications (1.7% eptifibatide group versus 0.7% abciximab group; p = 0.2) were similar between the 2 groups. Thrombocytopenia was more frequent in the abciximab-treated patients (6%, versus 0% in the eptifibatide group; p < 0.001), including 5 patients who developed profound thrombocytopenia (< 20,000 cells/mm3). CONCLUSION: Both agents, eptifibatide and abciximab, proved to have the same rates of in-hospital major adverse cardiac events, bleeding and vascular complications. Abciximab therapy was associated with a significantly higher incidence of thrombocytopenia within 4 hours of drug initiation, which prompted immediate drug discontinuation, but was not associated with increased risk of bleeding, vascular or other complications. PMID- 12777671 TI - Glycoprotein Wars and PIECE activists. PMID- 12777672 TI - Parameters of left ventricular diastolic function 48 hours after coronary angioplasty and stent implantation. AB - INTRODUCTION: It has been reported that stent implantation results in an earlier and more pronounced improvement of coronary flow reserve in comparison to conventional balloon angioplasty. Whether this phenomenon translates into hemodynamic changes of left ventricular systolic and diastolic function has not been investigated. This study was designed to determine whether stenting leads to greater changes in measures of diastolic dysfunction than plain angioplasty alone. METHODS: Parameters of diastolic function were ascertained by Doppler echocardiography in 194 patients with single-vessel disease before and 48 hours after elective coronary angioplasty. A total of 116 patients were initially successfully treated with coronary angioplasty. In 78 patients, stents were used to improve an inadequate result after coronary angioplasty. The parameters of left ventricular diastolic function were evaluated before and 48 hours after coronary intervention by Doppler echocardiography. Ejection fraction was determined and used to characterize systolic left ventricular function. RESULTS: Both patient groups (116 patients with coronary angioplasty, 78 patients with combined coronary angioplasty and stent implantation) showed no relevant differences concerning sex, age, atherosclerotic risk factors, exercise capacity and results of exercise electrocardiography. All patients who underwent stent implantation showed an early improvement of left ventricular diastolic function 48 hours after intervention. Surprisingly, there was no significant short-term improvement (48 hours) of diastolic function in patients with initially successful angioplasty. CONCLUSION: Stent implantation results in improved left ventricular diastolic function in comparison to conventional balloon angioplasty. This has to be attributed to a more immediate and increased antiischemic effectiveness due to the scaffolding properties of stents. PMID- 12777673 TI - Bivalirudin as a foundation anticoagulant in peripheral vascular disease: a safe and feasible alternative for renal and iliac interventions. AB - BACKGROUND: Many heparin (UFH) limitations are overcome by bivalirudin (Angiomax ). The pharmacokinetic profile of bivalirudin appears well suited for percutaneous peripheral intervention (PPI), yet few data exist regarding its safety and feasibility in this setting. METHODS: One hundred and eighty renal and 75 iliac PPIs performed between May 2001 and June 2002 with bivalirudin as anticoagulation were compared to a historical UFH control. Variables evaluated included thrombotic events, intracranial bleeding, major surgical complications, sheath removal time, vascular access complication, time to ambulate and length of stay (LOS). Follow-up included 6-month renal and iliac duplex ultrasound and ankle-brachial index. RESULTS: Procedural success was achieved in 100% of patients treated with bivalirudin, with no thrombotic events, intracranial bleeding or major surgical complications observed. Procedural success was achieved in 179/180 (99%) renal and 74/75 (98.6%) iliac patients treated with UFH. Significant differences were observed for sheath removal time < 60 minutes (84% versus 59%; p < 0.0001), time to ambulation < 6 hours (75.5% versus 58%; p < 0.0005) and LOS < 24 hours (85.5% versus 72%; p = 0.002) in bivalirudin-treated renal PPI patients versus UFH-treated patients, respectively. Significant differences were also observed in favor of bivalirudin for the iliac PPIs for sheath removal time < 60 minutes (p = 0.012) and time to ambulation < 6 hours (p = 0.039). Following 6-month renal and iliac duplex ultrasound, repeat PPI was required in 7/180 (3.9%) and 9/180 (5%) of renal, and 3/75 (4%) and 4/75 (5.3%) of iliac patients treated with bivalirudin or UFH, respectively. CONCLUSION: Bivalirudin is a safe and feasible alternative anticoagulant in renal and iliac PPI and may offer decreased sheath removal time, time to ambulation and LOS. A larger prospective randomized multicenter trial is warranted. PMID- 12777674 TI - Patients at low risk for periprocedural myocardial infarction can be identified by assessment immediately following percutaneous coronary intervention. AB - Despite utilizing optimal anticoagulant therapy during percutaneous coronary intervention (PCI), the incidence of periprocedural myocardial infarction (PPMI) remains 5 7% and evaluation of preprocedural clinical/angiographic characteristics has failed to reliably predict the likelihood of a PPMI. We hypothesized that immediate post-PCI assessment could identify a group of patients at very low risk for PPMI. A consecutive series of 258 PCI patients was stratified into 3 groups based on immediate post-PCI assessment. Group I (PPMI not expected) included those with an acceptable angiographic result of treated vessel (residual stenosis < 50%), TIMI 3 flow and absence of any intraprocedural complications. Group II (PPMI not unexpected) included those with an acceptable angiographic result, TIMI 3 flow but with any/all of the following: saphenous vein graft (SVG) PCI, transient closure of culprit vessel or major sidebranch, intracoronary thrombus, prolonged chest pain, electrocardiographic (ECG) changes, hypotension, resolved slow flow/no reflow, bailout glycoprotein IIb/IIIa inhibitor use, loss of a minor sidebranch or any angiographic residual stenosis > 50% with TIMI 3 flow. Group III (PPMI expected) included those with any angiographic result of native coronary artery or SVG with < TIMI 3 flow, unresolved chest pain, hypotension or ECG changes at the end of the PCI, loss of a major sidebranch or vessel, or persistent no-reflow. Group stratification was analyzed in relation to the incidence of PPMI (CK-MB > 3 times the upper limit of normal; 18 24 hours post-PCI). Rate of PPMI: Group I (1/141; 0.7%), Group II (7/71; 9.9%), Group III (5/11; 45.5%) (p < 0.001). The 3 groups did not differ in age, clinical presentation or stent use (p = NS). Sixty out of 105 patients (57.1%) with unstable angina, seventy-seven out of 146 patients (52.7%) with B2/C lesions, and 105/180 patients (58.3%) with unstable angina or B2/C lesions were stratified to Group I. This study demonstrates that immediate post-PCI evaluation of the clinical/angiographic characteristics can predict the likelihood of PPMI and a group of patients at a very low risk for a PPMI can be identified, in whom implications exist for limited hospitalization and post-procedural antithrombotic therapy. PMID- 12777675 TI - Dual-catheter covered stenting: a novel approach to the treatment of large coronary artery perforations. AB - Coronary perforation is an infrequent but potentially devastating complication of coronary intervention. Treatment of a broad-based perforation usually requires emergent pericardiocentesis and thoracotomy. We present a novel approach to closure of > or = 1 mm diameter perforations using a dual-catheter covered stent technique. PMID- 12777676 TI - Extraction of the radial artery during transradial coronary angiography: an unusual complication. AB - The transradial approach is currently an accepted alternative for vascular access during percutaneous coronary interventions. Access-site complications, such as mild hematoma, hematic effusions, and reduced or absent radial pulse, have been reported. We report the occurrence of total extraction of the radial artery during sheath removal. The occurrence of this complication emphasizes the need for meticulous attention and prudence when a patient complains of local pain during sheath extraction. PMID- 12777677 TI - Diffuse coronary artery spasm induced by guidewire insertion. AB - Coronary artery spasm is reported to occur in 1 5% of percutaneous coronary interventions. However, coronary spasm induced solely by guidewire insertion is very rare. We present a case of diffuse coronary artery spasm induced by guidewire insertion, which responded promptly to intracoronary nitrates. The case illustrates the importance of the recognition of this phenomenon for the selection of the optimal balloon and stent size. PMID- 12777678 TI - A totally occluded right coronary artery presenting with a normal electrocardiogram. AB - Total occlusion of the right coronary artery (RCA) typically manifests as ST segment elevation in the inferior leads and sometimes the lateral precordial leads of the surface electrocardiogram (ECG). We report a case of a patient with a normal electrocardiogram on presentation who, on angiography, revealed a totally occluded proximal RCA. Emergency angioplasty and stenting was successfully able to recanalize the entire RCA and restore TIMI III flow. ECGs performed post-procedure showed minimal change. The existence of a subendocardial microvascular network may have allowed this patient to escape what typically would have been a large ST-elevation inferior myocardial infarction. PMID- 12777679 TI - Percutaneous intervention on the LIMA: tackling the tortuosity. AB - Percutaneous intervention on a tortuous left internal mammary artery (LIMA) graft is often challenging as the LIMA develops straightening and accordioning after the guidewire and dilatation balloon are placed in the graft, making angiographic assessment of the result impossible if there is no flow through the graft. We describe a new technique that overcomes this problem. PMID- 12777680 TI - Mechanisms and methods to resolve edge effect. AB - Vascular brachytherapy (VBT) has established itself as a viable modality to treat in-stent restenosis (ISR). The problems associated with VBT have been understood well and remedied. Late thrombosis has been overcome to a great extent by prolonged antiplatelet therapy. Edge effect is another important limitation of VBT and is due to inadequate radiation coverage of the edges following VBT. It may be overcome by confining injury to the lesion segment and extending the radiation sources by a few millimeters from the injured segment. PMID- 12777681 TI - Complete heart block due to lyme carditis. AB - Lyme carditis is becoming a more frequent complication of Lyme disease, primarily due to the increasing incidence of this disease in the United States. Cardiovascular manifestations of Lyme disease often occur within 21 days of exposure and include fluctuating degrees of atrioventricular (AV) block, acute myopericarditis or mild left ventricular dysfunction and rarely cardiomegaly or fatal pericarditis. AV block can vary from first-, second-, third-degree heart block, to junctional rhythm and asystolic pauses. Patients with suspected or known Lyme disease presenting with cardiac symptoms, or patients in an endemic area presenting with cardiac symptoms with no other cardiac risk factors should have a screening electrocardiogram along with Lyme titers. We present a case of third-degree AV block due to Lyme carditis, illustrating one of the cardiac complications of Lyme disease. This disease is usually self-limiting when treated appropriately with antibiotics, and does not require permanent cardiac pacing. PMID- 12777682 TI - Regulation of neuronal proliferation and differentiation by nitric oxide. AB - Many studies have revealed the free radical nitric oxide (NO) to be an important modulator of vascular and neuronal physiology. It also plays a developmental role in regulating synapse formation and patterning. Recent studies suggest that NO may also mediate the switch from proliferation to differentiation during neurogenesis. Many mechanisms of this response are conserved between neuronal precursor cells and the cells of the vascular system, where NO can inhibit the proliferative response of endothelial and smooth-muscle cells to injury. In cultured neuroblastoma cells, NO synthase (NOS) expression is increased in the presence of various growth factors and mitogens. Subsequent production of NO leads to cessation of cell division and the acquisition of a differentiated phenotype. The inhibitory action of NO on neuroblast proliferation has also been demonstrated in vivo for vertebrate and invertebrate nervous systems, as well as in the adult brain. Potential downstream effectors of NO include the second messenger cyclic GMP, activation of the tumor-suppressor genes p53 and Rb, and the cyclin-dependent kinase inhibitor p21. These studies highlight a new role for NO in the nervous system, as a coordinator of proliferation and patterning during neural development and adult neurogenesis. PMID- 12777683 TI - Stress and the developing hippocampus: a double-edged sword? AB - The mechanisms that regulate neuronal function are a sum of genetically determined programs and experience. The effect of experience on neuronal function is particularly important during development, because early-life positive and adverse experience (stress) may influence the still "plastic" nervous system long term. Specifically, for hippocampal-mediated learning and memory processes, acute stress may enhance synaptic efficacy and overall learning ability, and conversely, chronic or severe stress has been shown to be detrimental. The mechanisms that enable stress to act as this "double-edged sword" are unclear. Here, we discuss the molecular mediators of the stress response in the hippocampus with an emphasis on novel findings regarding the role of the neuropeptide known as corticotropin-releasing hormone (CRH). We highlight the physiological and pathological roles of this peptide in the developing hippocampus, and their relevance to the long-term effects of early-life experience on cognitive function during adulthood. PMID- 12777684 TI - Physiological and anatomical link between Parkinson-like disease and REM sleep behavior disorder. AB - Parkinson's disease (PD) is a progressive neurodegenerative disease that is caused by a loss of neurons in the ventral midbrain. Parkinsonian patients often experience insomnia, parasomnias, and daytime somnolence. REM sleep behavior disorder (RBD) is characterized by vigorous movements during REM sleep, and may also be caused by neuronal degeneration in the central nervous system (CNS); however, the site of degeneration remains unclear. Both Parkinsonism and RBD become more prevalent with aging, with onset usually occurring in the sixties. Recent findings show that many individuals with RBD eventually develop Parkinsonism. Conversely, it is also true that certain patients diagnosed with Parkinsonism subsequently develop RBD. Postmortem examination reveals that Lewy bodies, Lewy neurites, and alpha-synuclein are found in brainstem nuclei in both Parkinsonism and RBD patients. In this article, we will discuss evidence that Parkinsonism and RBD are physiologically and anatomically linked, based on our animal experiments and other studies on human patients. PMID- 12777685 TI - Brain repair and neuroprotection by serum insulin-like growth factor I. AB - The existence of protective mechanisms in the adult brain is gradually being recognized as an important aspect of brain function. For many years, self-repair processes in the post-embryonic brain were considered of minor consequence or nonexistent. This notion dominated the study of neurotrophism. Thus, although the possibility that neurotrophic factors participate in brain function in adult life was prudently maintained, the majority of the studies on the role of trophic factors in the brain were focused on developmental aspects. With the recent recognition that the adult brain keeps a capacity for cell renewal, although limited, a new interest in the regenerative properties of brain tissue has emerged. New findings on the role of insulin-like growth factor I (IGF-I), a potent neurotrophic peptide present at high levels in serum, may illustrate this current trend. Circulating IGF-I is an important determinant of proper brain function in the adult. Its pleiotropic effects range from classical trophic actions on neurons such as housekeeping or anti-apoptotic/ pro-survival effects to modulation of brain-barrier permeability, neuronal excitability, or new neuron formation. More recent findings indicate that IGF-I participates in physiologically relevant neuroprotective mechanisms such as those triggered by physical exercise. The increasing number of neurotrophic features displayed by serum IGF-I reinforces the view of a physiological neuroprotective network formed by IGF-I, and possibly other still uncharacterized signals. Future studies with IGF-I, and hopefully other neurotrophic factors, will surely reveal and teach us how to potentiate the self-reparative properties of the adult brain. PMID- 12777686 TI - Microglial functions and proteases. AB - There is accumulating evidence that intracellular and extracellular proteases of microglia contribute to various events in the central nervous system (CNS) through both nonspecific and limited proteolysis. Cathepsin E and cathepsin S, endosomal/lysosomal proteases, have been shown to play important roles in the major histocompatibility complex (MHC) class II-mediated antigen presentation of microglia by processing of exogenous antigens and degradation of the invariant chain associated with MHC class II molecules, respectively. Some members of cathepsins are also involved in neuronal death after secreted from microglia and clearance of phagocytosed amyloid- beta peptides. Tissue-type plasminogen activator, a serine protease, secreted from microglia participates in neuronal death, enhancement of N-methyl-D-aspartate receptor-mediated neuronal responses, and activation of microglia via either proteolytic or nonproteolytic activity. Calpain, a calcium-dependent cysteine protease, has been shown to play a pivotal role in the pathogenesis of multiple sclerosis by degrading myelin proteins extracellulary. Furthermore, matrix metalloproteases secreted from microglia also receive great attention as mediators of inflammation and tissue degradation through processing of pro-inflammatory cytokines and damage to the blood-brain barrier. The growing knowledge about proteolytic events mediated by microglial proteases will not only contribute to better understanding of microglial functions in the CNS but also may aid in the development of protease inhibitors as novel neuroprotective agents. PMID- 12777687 TI - Functional peptide sequences derived from extracellular matrix glycoproteins and their receptors: strategies to improve neuronal regeneration. AB - Peptides derived from extracellular matrix proteins have the potential to function as potent therapeutic reagents to increase neuronal regeneration following central nervous system (CNS) injury, yet their efficacy as pharmaceutical reagents is dependent upon the expression of cognate receptors in the target tissue. This type of codependency is clearly observed in successful models of axonal regeneration in the peripheral nervous system, but not in the normally nonregenerating adult CNS. Successful regeneration is most closely correlated with the induction of integrins on the surface of peripheral neurons. This suggests that in order to achieve optimal neurite regrowth in the injured adult CNS, therapeutic strategies must include approaches that increase the number of integrins and other key receptors in damaged central neurons, as well as provide the appropriate growth-promoting peptides in a "regeneration cocktail." In this review, we describe the ability of peptides derived from tenascin- C, fibronectin, and laminin-1 to influence neuronal growth. In addition, we also discuss the implications of peptide/receptor interactions for strategies to improve neuronal regeneration. PMID- 12777690 TI - A simple PCR/RFLP analysis can differentiate between Candida albicans, Aspergillus niger, and Aspergillus fumigatus. AB - The clinical management of immunocompromised patients depends on the rapid identification of infectious agents such as fungal pathogens. The procedure described here for accomplishing this uses a sensitive polymerase chain reaction method, previously reported, combined with restriction-enzyme digestion to distinguish between Candida and Aspergillus species and to classify Aspergillus strains. PMID- 12777689 TI - Molecular mechanisms that regulate auditory hair-cell differentiation in the mammalian cochlea. AB - Mechanosensory hair cells of the vertebrate cochlea offer an excellent developmental system to study cell-fate specification, and to gain insight into the many human neurological deficits which result in a hearing loss, by affecting primarily the hair cells. Therefore, there is great interest in studying the molecular mechanisms that regulate their specification and differentiation. Recent studies, based mostly on loss-of-function experiments that target the role of Notch signaling and basic helix-loop-helix genes in inner-ear development have indicated that they can regulate mechanosensory hair cell-fate specification and their initial differentiation. PMID- 12777691 TI - Rapid single-base mismatch detection in genotyping for phenylketonuria. AB - Phenylketonuria (PKU) is a metabolic disorder that results from a deficiency of hepatic phenylalanine hydroxylase (PAH). Identification of the PKU genotype is useful for predicting clinical PKU phenotype. More than 400 mutations resulting in PAH deficiency have been reported worldwide. We used a genedetecting instrument to identify the nine prevalent Japanese mutations in the PAH gene among 31 PKU patients as a preliminary study. This instrument can automatically detect mutations through the use of allelespecific oligonucleotide (ASO) capture probes, and gave results comparable to those of sequencing studies. Each country has uniquely prevalent and specific mutations causing PKU, and less than 50 types of such mutations are generally present in each country. Early genotyping of PKU makes it possible to identify the phenotype and select the optimal therapy for the disease. For early genotyping, the instrumental method described here shortens the time required for genotyping based on mRNA and/or genomic DNA of PKU parents. PMID- 12777688 TI - Visualizing activation of opioid circuits by internalization of G protein-coupled receptors. AB - Mu-opioid receptor (MOR) and opioid receptor-like receptor (ORL-1) circuits in the limbic hypothalamic system are important for the regulation of sexual receptivity in the female rat. Sexual receptivity is tightly regulated by the sequential release of estrogen and progesterone from the ovary suggesting ovarian steroids regulate the activity of these neuropeptide systems. Both MOR and ORL-1 distributions overlap with the distribution of estrogen and progesterone receptors in the hypothalamus and limbic system providing a morphological substrate for interaction between steroids and the opioid circuits in the brain. Both MOR and ORL-1 are receptors that respond to activation by endogenous ligands with internalization into early endosomes. This internalization is part of the mechanism of receptor desensitization or down regulation. Although receptor activation and internalization are separate events, internalization can be used as a temporal measure of circuit activation by endogenous ligands. This review focuses on the estrogen and progesterone regulation of MOR and ORL-1 circuits in the medial preoptic nucleus and ventromedial nucleus of the hypothalamus that are central to modulating sexual receptivity. PMID- 12777692 TI - What would T. H. Huxley have made of prion diseases? AB - T. H. Huxley was "Darwin's bulldog," and took the offensive in championing the cause of evolution against skeptical scientists and outraged theologians. As such, he took part in one of the great "paradigm shifts" of biology, at the end of the nineteenth century. Huxley was a rigorous scientist and wrote important articles on scientific method, as well as publishing extensively on a wide range of subjects in natural history. In the second half of the twentieth century, the "prion hypothesis" was put forward to explain the pathogenesis of a curious group of diseases known as the transmissible spongiform encephalopathies. This also involved a "paradigm shift" because the prion hypothesis postulated that biologically relevant information could be enciphered in protein conformation (rather than encoded in nucleic acid base sequences), and could be transmitted from one molecule to another, thereby causing infectious disease. This article examines a few of Huxley's remarks to speculate on how he might have responded to the scientific debate about prion disease had he lived a century later. PMID- 12777694 TI - Renal gene transfer: nonviral approaches. AB - Gene therapy has the potential to become an important modality for treating both hereditary and acquired renal diseases. Since renal diseases may involve different cell types in the kidney, it is critical to achieve efficient gene transfer specifically to each cell type. We reviewed the literature on nonviral gene transfer techniques, which are designed to target the kidney specifically. A variety of approaches have been developed to target glomeruli, tubules, renal vasculature, and interstitium with different degree of success. Besides using delivery systems based on liposomes, polycations, and viral fusion proteins, investigators have adopted newer approaches including electroporation and hydrodynamic-based gene transfer, and demonstrated that they are efficient and safe in animal models. Potential clinical applications and safety concerns of gene therapy for renal diseases are discussed. PMID- 12777693 TI - A new look at xylanases: an overview of purification strategies. AB - Interest in xylanases from different sources has increased markedly in the past decade, in part because of the application of these enzymes in the pulp and paper industry. Purity and purification costs are becoming important issues in modern biotechnology as the industry matures and competitive products reach the marketplace. Thus, new paths for successful and efficient xylanase recovery have to be followed. This article reviews the isolation and purification methods used for the recovery of microbial xylanases. Origins and applications of xylanases are described, highlighting the special features of this class of enzymes, such as the carbohydrate-binding domains (CBDs) and their importance in the development of affinity methodologies to increase and facilitate xylanase purification. Implications of recombinant DNA technology for the isolation and purification of xylanases are evaluated. Several purification procedures are analyzed, taking into consideration the sequence of the methods used in each and the number of times each method is used. New directions to improve xylanase separation and purification from fermentation media are described. PMID- 12777695 TI - An introduction to telomeres and telomerase. AB - It is now more than a dozen years since the enzyme telomerase was discovered, and since that time, key studies have characterized the structural components of the enzyme and the associated telomeric proteins. Since the original discovery of telomerase, a clear association with cancer has been demonstrated. In normal somatic cells the telomeres at the ends of chromosomes shorten with every cell division, whereas in cancer cells telomere length is often maintained by reactivation of the enzyme telomerase. These discoveries have led to the proposal that telomerase expression can be used as a helpful marker for diagnostic and prognostic purposes in humans. Another area of research that has developed as a result of improving knowledge and understanding of the role of telomerase in malignancy is that of cancer therapeutics. This article is an introduction to the field of telomere and telomerase research, with an introduction to recent attempts to develop novel cancer treatments based on telomerase structure and function. PMID- 12777696 TI - Automated pneumococcal MLST using liquid-handling robotics and a capillary DNA sequencer. AB - Multilocus sequence typing (MLST) is used by the Scottish Meningococcus and Pneumococcus Reference Laboratory (SMPRL) as a routine method for the characterization of certain bacterial pathogens. The SMPRL recently started performing MLST on strains of Streptococcus pneumoniae, and here we describe a fully automated method for MLST using a 96-well-format liquid-handling robot and a 96-capillary automated DNA sequencer. PMID- 12777697 TI - Cellular strategies of estrogen-mediated neuroprotection during brain development. AB - The role of estrogen during brain development is well documented. Estrogen influences cell survival and differentiation and also controls the formation and maintenance of neural networks. Knowledge of trophic estrogen action in the central nervous system (CNS) was the basis for the establishment of research programs directed toward a potential function of estrogen as a neuroprotective factor in the adult brain. Considerable evidence has accumulated over the years supporting this hypothesis. Experimental and epidemiologic studies as well as clinical trials have demonstrated that estrogen is beneficial for the course of neurodegenerative disorders such as Parkinson and Alzheimer diseases but may also protect neurons from postischemic neuronal degeneration. In this article, we aim to unravel potential physiologic responses and cell survival strategies that allow a more detailed understanding of estrogen-mediated neuroprotection in the brain. In particular, we focus on the participation of estrogen in the regulation of apoptotic processes. Furthermore, we present data on reciprocal estrogen growth factor interactions. Both of these mechanisms were found to operate during brain development and to conciliate estrogen effects on neurons. This makes them likely candidates for taking part in conveying estrogen-dependent neuroprotection in the adult CNS. PMID- 12777698 TI - Estradiol exerts neuroprotective actions against ischemic brain injury: insights derived from animal models. AB - Over the last 100 yr, the life-span of women has increased from 50 yr to over 80 yr, but the age of the menopause has remained unchanged, at 51 yr. Menopause is one of the most permanent physiologic changes that a woman will experience and is marked by a dramatic decrease in circulating levels of ovarian estrogens. Because the timing of menopause has remained fixed in the face of an increasing life span, more women will live a greater proportion of their lives in a hypoestrogenic state. We appreciate more and more that the actions of ovarian steroid hormones are complex, and possibly exert opposing actions in different contexts. I review here the results of my laboratory's recent studies that clearly establish that low physiologic levels of estradiol replacement can exert profound neuroprotective actions when administered prior to an ischemic strokelike injury. PMID- 12777699 TI - Estrogen and selective estrogen receptor modulators: neuroprotection in the Women's Health Initiative era. AB - Estrogen has been comprehensively studied as a neuroprotective agent in women, animals, and a variety of in vitro models of neural injury and degeneration. Most data suggest that estrogen can benefit the ischemic brain and reduce cell death. However, recent data from the Women's Health Initiative have raised concerns about the utility and safety of chronic estrogen use in women. While estrogen is a potent and reproducible neuroprotectant in animals and in vitro, its current administration in women has had unanticipated and paradoxical effects. Nonetheless, estrogen's diverse actions make it an ideal prototype for developing new neuroprotectants such as selective estrogen receptor modulators (SERMs). SERMs represent a class of drugs with mixed estrogen agonistic and antagonistic activity. Experimental and clinical data suggest a neuroprotective role for SERMs in normal and injured brain. The discrepancy among observational studies, preclinical data, and clinical trials emphasizes the need for further study of the mechanisms leading to the increased incidence of stroke observed in postmenopausal women. Research is still needed to optimize combined or estrogen alone hormone replacement therapy options as well as the prevention/management of cerebrovascular/ central nervous system disorders. This review critiques estrogen and SERMs' neuroprotective potential in experimental and clinical studies of stroke and cerebrovascular disease. PMID- 12777700 TI - Induction of antioxidative and antiapoptotic thioredoxin supports neuroprotective hypothesis of estrogen. AB - The original neuroprotective hypothesis of estrogen was based on the gender difference in brain response to the ischemia-reperfusion injury. Additional clinical reports also suggest that estrogen may improve cognition in patients with Alzheimer disease. 17beta-Estradiol is the most potent endogenous ligand of estrogen, which protects against neurodegeneration in both cell and animal models. Estrogen-mediated neuroprotection is probably mediated by both receptor dependent and -independent mechanisms. Binding of estrogen such as 17beta estradiol to estrogen receptors (ERs) activates the homodimers of ER-DNA and its binding to estrogen response elements in the promoter region of genes such as neuronal nitric oxide synthase (NOS1) for regulating gene expression in target brain cells. In addition to the induction of NOS1, estrogen increases the expression of antiapoptotic protein such as bcl-2. Furthermore, our recent observations provide new molecular biologic and pharmacologic evidence suggesting that physiologic concentrations of 17beta-estradiol (<10 nM) activate ERs (ERbeta > ERalpha) and upregulate a cyclic guanosine 5'- monophosphate (cGMP)-dependent thioredoxin (Trx) and MnSOD expression following the induction of NOS1 in human brain-derived SH-SY5Y cells. We thus proposed that the estrogen-mediated gene induction of Trx plays a pivotal role in the promotion of neuroprotection because Trx is a multifunctional antioxidative and antiapoptotic protein. For managing progressive neurodegeneration such as Alzheimer dementia, our estrogen proposal of the signaling pathway of cGMP-dependent protein kinase (PKG) in mediating estrogen-induced cytoprotective genes thus fosters research and development of the new estrogen ligands devoid of female hormonal side effects such as carcinogenesis. PMID- 12777701 TI - Functional aspects of estrogen neuroprotection. AB - Evidence that estrogen protects neurons against toxic/ ischemic insults or degenerative/aging processes is evident in a variety of in vitro and in vivo systems. However, a critical remaining question is: Does the demonstrated morphologic and neurochemical protection by estrogen lead to a preservation of brain function or an enhanced ability to recover? To date, little basic research is available on this issue. Cognition is a critical function that might provide a sensitive way to examine this question. As a first step, we present results showing that two chronic environmental insults, psychoactive drugs and stress, produce gender-specific responses in cognitive abilities. Specifically, females appear less sensitive than males to cognitive impairments following chronic exposure to these factors. Results are presented in male and female rats utilizing cognitive tests that assess visual (object recognition) and spatial memory (object placement and radial arm maze) following chronic amphetamine, methamphetamine, or daily restraint stress. Following regimes of chronic stress or amphetamine, males were impaired on these tasks while females were either unaffected, less affected, or enhanced in performance. These observations suggest that differences in circulating gonadal hormone levels between the sexes may contribute to the differential sensitivity of the sexes and provide endogenous neuroprotection for females. Surprisingly, ovariectomized females were still not impaired following a stress regimen that impaired males (21 d of daily restraint). These data taken together with neurochemical data on estrogen neuroprotective effects indicate that it is possible that neuroprotection by estrogen may result from hormone action both during sexual differentiation (organizational effect) and in adulthood (activational effect). These considerations and possible unwanted/untoward effects of chronic estrogen use are discussed in relation to the use of selective estrogen receptor modulators for chronic treatment of both males and females. In conclusion, although compelling evidence for neuroprotection by estrogen has been presented in anatomic and neurochemical studies, it is clear that the functional/ behavioral aspects need further investigation. PMID- 12777702 TI - Endogenous estrogen formation is neuroprotective in model of cerebellar ataxia. AB - The expression of aromatase, the enzyme that transforms testosterone into estradiol, was analyzed by reverse transcriptase polymerase chain reaction in the inferior olive of adult male rats. The expression of this messenger in the inferior olive suggests that this brain area may be able to synthesize estradiol. The neuroprotective role of estradiol in the inferior olive was then assessed in a model of cerebellar ataxia, achieved by the ip administration of 3 acetylpyridine (3-AP). In a first experiment, male Wistar rats were orchidectomized to diminish the plasmatic levels of testosterone, the direct precursor of estradiol. Immediately after castration, animals were implanted with a silicone tube that was either empty or filled with estradiol. One week later, animals were injected with 3-AP. Estradiol treatment resulted in a significant reduction in neuronal death in the olive. In a second experiment, animals were treated with fadrozole, an aromatase inhibitor, to assess the role of endogenous estradiol formation in neuroprotection. The results show that the inhibition of aromatase activity, and therefore the decrease in endogenous estrogen formation, increases the death in inferior olive. In conclusion, this study indicates that the inferior olive is a steroidogenic tissue and that olivary neurons are protected by exogenous and endogenous estradiol. PMID- 12777703 TI - Chronic but not acute estradiol treatment protects against the neurodegenerative effects of N-methyl-D-aspartate receptor antagonists. AB - Drugs that block NMDA receptors, thereby inducing an NMDA receptor hypofunctional (NRHypo) state, can cause a disseminated pattern of irreversible neurodegeneration. Based on several lines of evidence, an N-methyl-D-aspartate receptor hypofunction (NRHypo) mechanism has been postulated to contribute to neurodegenerative changes in Alzheimer disease (AD). Because estrogen putatively exerts a neuroprotective effect in AD, we examined whether estrogen protects against NRHypo-induced neurodegeneration. We administered estradiol benzoate in three separate experiments to adult female rats: (1) 100 microg subcutaneously as a onetime dose, (2) 100 microg bid twice daily for 4.5 or 14 d, and 3) 300 microg twice daily for 4.5 d. Two hours after the last estradiol dose, MK-801 was administered (0.5 mg/kg subcutaneously) to produce a robust neurotoxic injury. Controls received MK-801, but no estradiol. Four hours after administration of MK 801, the severity of injury was evaluated histologically by quantitative methods previously described. Compared to controls, a single dose of estradiol produced no change in the severity of injury (p = 0.24). Chronic treatment with estradiol was associated with a 25-35% reduction in the number of injured neurons (p < 0.05 in all cases). We conclude that chronic but not acute estradiol treatment reduces the severity of NRHypo-induced neurodegeneration. PMID- 12777704 TI - Neuroprotective effects of estrogen and tamoxifen in vitro: a facilitative role for glia? AB - Selective estrogen receptor modulators (SERMs) are steroidal or nonsteroidal compounds that can exhibit either estrogen-like agonistic effects or estrogen antagonistic effects depending on the target tissue. While SERM actions in the breast, bone, and uterus have been well characterized, their effects in the brain are considerably less well understood. Previous work by our laboratory has demonstrated a beneficial effect of tamoxifen in the reduction of ischemic stroke damage in ovariectomized female rats. The present study utilized neuronal cell culture models to attempt to understand the mechanisms of tamoxifen-mediated neuroprotection. Neither physiologic doses of 17beta-E2 nor clinically therapeutic doses of tamoxifen directly protected GT1-7 neurons or purified cultures of rat cerebrocortical neurons from several forms of cell death. Reverse transcriptase polymerase chain reaction and Western blot analysis revealed that GT1-7 neurons possessed both estrogen receptor-alpha (ERalpha) and ERbeta mRNA and protein, whereas purified embryonic rat cortical neurons only expressed appreciable levels of ERalpha transcript and protein, with little to no expression of ERbeta. In contrast to the lack of protection in the purified neuronal cultures, both 17beta- E2 and tamoxifen significantly protected mixed glial/ neuronal cortical cultures from cell death, suggesting that glia may facilitate 17beta-E2-and tamoxifen-mediated neuroprotection. Furthermore, astrocyte-conditioned media and exogenous transforming growth factor-beta1, a documented astrocyte-derived cytokine, were shown to rescue purified cortical neurons from cell death. Together, these findings support a role for astrocytes in neuroprotection and raise the intriguing possibility that astrocytes may help mediate the neuroprotective effect of 17beta-E2 and tamoxifen. PMID- 12777705 TI - Estrogen as neuroprotectant of nigrostriatal dopaminergic system: laboratory and clinical studies. AB - In this review, we relate both laboratory and clinical evidence associated with the capacity for estrogen to function as a modulator of nigrostriatal dopaminergic pathology. To accomplish this goal, we have divided this review into three parts. In Part 1, we provide a brief historical perspective of studies that have laid the groundwork for demonstrating the existence of hormonal- nigrostriatal interactions. In Part 2, we focus specifically on laboratory data that show the ability and conditions by which estrogen may function as a neuroprotectant of the nigrostriatal dopaminergic system. Finally, in Part 3, we review the clinical literature related to this issue as a means for consideration of estrogen as a modulator, neuroprotectant, and therapy for Parkinson disease. PMID- 12777706 TI - Estrogens and Parkinson disease: novel approach for neuroprotection. AB - Epidemiologic studies revealed that the prevalence of Parkinson disease is higher in males than in females and that the progression of the disease might be rapid in males compared with females. The reason for the gender difference is unknown; however, estrogens may be involved. Many studies have revealed that estrogens provide neuroprotective effects and that the protective mechanisms include antioxidant property and upregulation of Bcl-2, brain-derived neurotrophic factor, and glial cell-derived neurotrophic factor (GDNF). Upregulation of Bcl-2 or GDNF is mediated by nonnuclear estrogen receptor (ER) in addition to transcription regulation by ER. To avoid undesirable effect of estrogens, several selective ER modulators, raloxifene and genistein are considered. PMID- 12777707 TI - Estrogens and Parkinson disease: neuroprotective, symptomatic, neither, or both? AB - Parkinson disease is a neurodegenerative disorder caused by substantia nigra dopamine cell death and is characterized by bradykinesia, rigidity, rest tremor, and postural instability. Epidemiologic and clinical studies have suggested that gender and estrogen play a role in modulating Parkinson disease. The etiology of the estrogenic effect is unclear-it may be neuroprotective, symptomatic, or both. Retrospective studies suggest a possible neuroprotective role. Interventional studies have suggested a positive modulatory role or no role at all. While it is difficult to establish whether there is a true neuroprotective benefit of estrogen in the setting of even mild symptomatic benefit, laboratory data suggest such a neuroprotective role. Estrogen may act as an antiapoptotic agent, an antioxidant, or a neurotrophic modulating agent, promoting crosstalk with neurotrophic factors. The selective estrogen receptor modulators (SERMs) may also confer neuroprotection. However, prior to establishing the role of estrogen in Parkinson disease, additional study, including of the SERMs, is warranted. PMID- 12777709 TI - Estradiol and dehydroepiandrosterone potentiate levodopa-induced locomotor activity in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine monkeys. AB - Six monkeys were rendered hemiparkinsonian with a unilateral injection of 1 methyl-4-phenyl-1,2,3,6-tetrahydropyridine. These monkeys displayed ipsilateral circling under basal conditions, and after dopaminergic stimulation with levodopa they decreased their ipsilateral circling and started turning to the contralateral side of their lesion. The effect of 17beta-estradiol and dehydroepiandrosterone (DHEA) was investigated in these animals. 17beta-Estradiol (0.1 mg/kg) added to a threshold dose of levodopa significantly potentiated contralateral circling (mean/30 min) compared to saline or threshold levodopa treatment whereas the duration of circling remained unchanged. DHEA (1-15 mg/kg) alone induced contralateral circling, compared to saline treatment, for 90 min. In addition, DHEA (1-15 mg/kg) potentiated the contralateral circling (mean/30 min) induced by a threshold dose of levodopa and did not change the duration of levodopa circling. A maximal response was observed with 1 or 5 mg/kg of DHEA combined with levodopa depending on the monkey. No correlation was found between the dose for the maximal DHEA response and baseline circling or threshold dose of levodopa. These results suggest that 17beta-estradiol or DHEA is able to potentiate locomotor activity of hemiparkinsonian monkeys. The DHEA doses investigated are similar to those presently used in humans. DHEA may be an alternative to 17beta-estradiol to modulate dopaminergic activity. PMID- 12777708 TI - Effects of estradiol, phytoestrogens, and Ginkgo biloba extracts against 1-methyl 4-phenyl-pyridine-induced oxidative stress. AB - Oxidative stress has been recently considered as a mediator of nerve cell death in several neurodegenerative diseases. We studied the effect of the parkinsonism inducing toxine 1-methyl-4-phenyl-pyridine (MPP+) on several parameters of cell distress using native and neuronal PC12 cells. Then, since estrogens have been reported to prevent neuronal degeneration caused by oxidative damage, we investigated the ability of 17beta- estradiol (E2); two Ginkgo biloba extracts, EGb 761 and Cp 202; as well as two flavonoids, quercetin and kaempferol, to rescue PC12 cells submitted to MPP+- induced oxidative stress. Our results consistently show that both Ginkgo biloba extracts could prevent cell death in native and neuronal PC12 cells, while in neuronal PC12 cells also quercetin and E2 could reverse MPP+ neurotoxic effet. Western blot analysis demonstrated that MPP+ injuries might modulate dopamine transporter (DAT) protein expression but not estrogen receptor beta (ERbeta) protein expression. EGb 761 and Cp 202 also modulate DAT and ERbeta protein expression in neuronal cells. From these studies, we outline the importance of testing estrogen-like plant-derived molecules as potent antioxidants and examine their effect on protein expression. PMID- 12777710 TI - Coating and selective deposition of nanofilm on silicone rubber for cell adhesion and growth. AB - A recently developed method for surface modification, layer-by-layer (LbL) assembly, has been applied to silicone, and its ability to encourage endothelial cell growth and control cell growth patterns has been examined. The surfaces studied consisted of a precursor, with alternating cationic polyethyleneimine (PEI) and anionic sodium polystyrene sulfonate (PSS) layers followed by alternating gelatin and poly-D-lysine (PDL) layers. Film growth increased linearly with the number of layers. Each PSS/PEI bilayer was 3 nm thick, and each gelatin/PDL bilayer was 5 nm thick. All layers were more hydrophilic than the unmodified silicone rubber surface, as determined from contact angle measurements. The contact angle was primarily dictated by the outermost layer. Of the coatings studied, gelatin was the most hydrophilic. A film of (PSS/PEI)4/(gelatin/PDL)4/ gelatin was highly favorable for cell adhesion and growth, in contrast to films of (PSS/PEI)8 or (PSS/PEI)8/PSS. Cell growth patterns were successfully controlled by selective deposition of microspheres on silicone rubber, using microcontact printing with a silicone stamp. Cell adhesion was confined to the region of microsphere deposition. These results demonstrate that the LbL self-assembly technique provides a general approach to coat and selectively deposit films with nanometer thickness on silicone rubber. Furthermore, they show that this method is a viable technique for controlling cellular adhesion and growth. PMID- 12777711 TI - A serendipitous discovery of antifreeze protein-specific activity in C-linked antifreeze glycoprotein analogs. AB - Structurally diverse carbon-linked (C-linked) analogs of antifreeze glycoprotein (AFGP) have been prepared via linear or convergent solid phase synthesis. These analogs range in molecular weight from approx 1.5-4.1 KDa and do not possess the beta-D-galactose-1,3-alpha-D-N-acetylgalactosamine carbohydrate moiety or the L threonine-L-alanine-L-alanine polypeptide backbone native to the AFGP wild-type. Despite these dramatic structural modifications, the 2.7-KDa and 4.1-KDa analogs possess antifreeze protein-specific activity as determined by recrystallization inhibition (RI) and thermal hysteresis (TH) assays. These analogs are weaker than the wild-type in their activity, but nanoliter osmometry indicates that these compounds are binding to ice and affecting a localized freezing point depression. This is the first example of a C-linked AFGP analog that possesses TH and RI activity and suggests that the rational design and synthesis of chemically and biologically stable AFGP analogs is a feasible and worthwhile endeavor. Given the low degree of TH activity, these compounds may prove useful for the protection of cells during freezing and thawing cycles. PMID- 12777712 TI - Rat polymerase beta gapped DNA interactions: antagonistic effects of the 5' terminal PO4 - group and magnesium on the enzyme binding to the gapped DNAs with different ssDNA gaps. AB - The role of the 5' terminal phosphate group downstream from the primer and magnesium cations in the energetics and dynamics of the gapped DNA recognition by rat polymerase beta have been examined, using the fluorescence titration and stopped-flow techniques. The analyses have been performed with the entire series of gapped DNA substrates differing in the size of the ssDNA gap. The 5' terminal phosphate group and magnesium cations exert antagonistic effect on enzyme binding to gapped DNA that depends on the length of the ssDNA gap. The PO4 - group amplifies the differences between the substrates with different ssDNA gaps, while in the presence of magnesium, affinities and structural changes induced in the DNA are very similar among examined DNA substrates. Both, the phosphate group and Mg+2 differ dramatically in affecting the thermodynamic response of the gapped DNA-rat pol beta system to the salt concentration. The data indicate that these distinct effects result from affecting the structure of the DNA, in the case of the phosphate group, and from direct magnesium binding to the protein. The mechanism of rat enzyme binding depends on the length of the ssDNA gap and the presence of the 5' terminal phosphate group. Complex formation with DNAs having three, four, and five residues in the gap occurs by a minimum three-step sequential mechanism. Depending on the presence of the 5' terminal phosphate group and/or magnesium, binding of the enzyme to a DNA containing two residues in the ssDNA gap is described by the same three-step or by a simpler two-step mechanism. With the DNA containing only one residue in the gap, binding is always described by only a two-step mechanism. The PO4 - group and magnesium cations have opposite effects on internal stability of the complexes with different length of the ssDNA gap. While the PO4 - group increases the stability of internal intermediates with the increasing length of the gap, Mg+2 decreases the stability of the intermediates with longer ssDNA gap. As a result, the combined favorable orientation effect of the phosphate group and the unfavorable Mg+2 effect lead to the optimal docking of the ssDNA gaps with three and four residues by the enzyme. PMID- 12777713 TI - Effects of membrane lipids on ion channel structure and function. AB - Biologic membranes are not simply inert physical barriers, but complex and dynamic environments that affect membrane protein structure and function. Residing within these environments, ion channels control the flux of ions across the membrane through conformational changes that allow transient ion flux through a central pore. These conformational changes may be modulated by changes in transmembrane electrochemical potential, the binding of small ligands or other proteins, or changes in the local lipid environment. Ion channels play fundamental roles in cellular function and, in higher eukaryotes, are the primary means of intercellular signaling, especially between excitable cells such as neurons. The focus of this review is to examine how the composition of the bilayer affects ion channel structure and function. This is an important consideration because the bilayer composition varies greatly in different cell types and in different organellar membranes. Even within a membrane, the lipid composition differs between the inner and outer leaflets, and the composition within a given leaflet is both heterogeneous and highly dynamic. Differential packing of lipids (and proteins) leads to the formation of microdomains, and lateral diffusion of these microdomains or "lipid rafts" serve as mobile platforms for the clustering and organization of bilayer constituents including ion channels. The structure and function of these channels are sensitive to specific chemical interactions with neighboring components of the membrane and also to the biophysical properties of their membrane microenvironment (e.g., fluidity, lateral pressure profile, and bilayer thickness). As specific examples, we have focused on the K+ ion channels and the ligand-gated nicotinicoid receptors, two classes of ion channels that have been well-characterized structurally and functionally. The responsiveness of these ion channels to changes in the lipid environment illustrate how ion channels, and more generally, any membrane protein, may be regulated via cellular control of membrane composition. PMID- 12777714 TI - Brownian ratchet models of molecular motors. AB - Brownian ratchet theory refers to the phenomenon that nonequilibrium fluctuations in an isothermal medium and anisotropic system can induce mechanical force and motion. This concept of noise-induced transport has motivated an abundance of theoretical and applied research. One of the exciting applications of the ratchet theory lies in the possible explanation of the operating mode of biological molecular motors. Biomolecular motors are proteins able of converting chemical energy into mechanical motion and force. Because of their dimension, the many small parts that make up molecular motors must operate at energies only a few times greater than those of the thermal baths. The description of molecular motors must be stochastic in nature. Here, we review the theoretical concepts of the Brownian ratchet theory and its possible link to the operation of biomolecular motors. We illustrate the principle of the ratchet theory with models of two molecular motors: a rotary motor (F0F1ATP synthase) and a linear motor (myosin II). PMID- 12777715 TI - Voltage-gated sodium channel toxins: poisons, probes, and future promise. AB - Neurotoxins have served as invaluable agents for identification, purification, and functional characterization of voltage-gated ion channels. Multiple classes of these toxins, which target voltage- gated Na+ channels via high-affinity binding to distinct but allosterically coupled sites, have been identified. The toxins are chemically diverse, including guanidinium heterocycles, a variety of structurally unrelated alkaloids, and multiple families of nonhomologous polypeptides having either related or distinct functions. This review describes the biochemistry and pharmacology of these agents, and summarizes the structure function relationships underlying their interaction with molecular targets. In addition, we explore recent advances in the use of these toxins as molecular scaffolding agents, drugs, and insecticides. PMID- 12777717 TI - Generation and application of phospho-specific antibodies for p53 and pRB. PMID- 12777716 TI - Utilizing NMR to study the structure of growth-inhibitory proteins. PMID- 12777718 TI - Stability and ubiquitination of the tumor suppressor protein p53. PMID- 12777719 TI - Role of tumor suppressors in DNA damage response. PMID- 12777720 TI - In situ hybridization in cancer and normal tissue. PMID- 12777721 TI - Genetic strategies in Saccharomyces cerevisiae to study human tumor suppressor genes. PMID- 12777722 TI - Electrophoretic mobility shift analysis of the DNA binding of tumor suppressor gene products. PMID- 12777723 TI - Analysis of gene promoter regulation by tumor suppressor genes. PMID- 12777724 TI - DNA footprinting. PMID- 12777725 TI - Identification of DNA-binding of tumor suppressor genes by chromatin immunoprecipitation. PMID- 12777726 TI - Co-immunoprecipitation of tumor suppressor protein-interacting proteins. PMID- 12777727 TI - Microarray approaches for analysis of tumor suppressor gene function. PMID- 12777728 TI - Analysis of tumor suppressor gene-induced senescence. PMID- 12777729 TI - Yeast two-hybrid screening as a means of deciphering tumor suppressor pathways. PMID- 12777730 TI - Somatic cell knockouts of tumor suppressor genes. PMID- 12777731 TI - Colony growth suppression by tumor suppressor genes. PMID- 12777732 TI - Flow cytometric analysis of cell cycle control by tumor suppressor genes. PMID- 12777733 TI - Analysis of cyclin-dependent kinase activity. PMID- 12777734 TI - Tumor suppressor gene-inducible cell lines. PMID- 12777735 TI - In vitro models of early neoplastic transformation of human mammary epithelial cells. PMID- 12777737 TI - Flow cytometric analysis of tumor suppressor gene-induced apoptosis. PMID- 12777736 TI - Tumor suppression through angiogenesis inhibition. PMID- 12777738 TI - Functional analysis of tumor suppressor genes in mice. PMID- 12777739 TI - The tissue microenvironment as an epigenetic tumor modifier. PMID- 12777740 TI - Analyzing the function of tumor suppressor genes using a Drosophila model. PMID- 12777741 TI - Assembling a tumor progression model. PMID- 12777742 TI - Conformation-sensitive gel electrophoresis for detecting BRCA1 mutations. PMID- 12777743 TI - Conversion technology and cancer predispositions. PMID- 12777744 TI - Discovering novel anticancer drugs: practical aspects and recent advances. PMID- 12777745 TI - Targets in apoptosis signaling: promise of selective anticancer therapy. PMID- 12777746 TI - Tumor deprivation of oxygen and tumor suppressor gene function. PMID- 12777747 TI - Hormonal and differentiation agents in cancer growth suppression. PMID- 12777748 TI - Regulation of NF-kappaB by oncoproteins and tumor suppressor proteins. PMID- 12777749 TI - Blocking survivin to kill cancer cells. PMID- 12777750 TI - Targeting the mitochondria to enhance tumor suppression. PMID- 12777751 TI - Novel approaches to screen for anticancer drugs using Saccharomyces cerevisiae. PMID- 12777752 TI - Tumor suppressor gene therapy. PMID- 12777753 TI - Therapeutic strategies using inhibitors of angiogenesis. PMID- 12777754 TI - Isolation of p53 inhibitors by screening chemical libraries in cell-based readout system. PMID- 12777756 TI - Structure determination of adeno-associated virus 2: three complete virus particles per asymmetric unit. AB - The atomic structure of adeno-associated virus 2 (AAV-2) has been determined to 3.0 A resolution. AAV-2 crystallized in space group P1, with unit-cell parameters a = 249.7, b = 249.7, c = 644.8 A, alpha = 90.0, beta = 101.2, gamma = 120.0 degrees. The crystals contained three full virus particles in the asymmetric unit, allowing 180-fold non-crystallographic symmetry averaging. The particle orientations were determined using the self-rotation function and found to have similar but resolvably different orientations. Approximate alignment of icosahedral and interparticle threefold screw symmetry led to a native Patterson that was interpretable in terms of approximate particle positions. Accurate positions required a Patterson correlation search that was constrained to be consistent with non-crystallographic threefold projection symmetry evident in the diffraction intensities. Initial phases to 15.0 A resolution were calculated by molecular replacement using the known structure of a distantly related homolog (23% sequence identity). Real-space averaging was performed and phases were extended from 15.0 to 3.0 A. An atomic model was fitted and refined using a simulated-annealing real-space procedure. PMID- 12777757 TI - High-resolution structure of human phosphoserine phosphatase in open conformation. AB - The crystal structure of human phosphoserine phosphatase (HPSP) in the open conformation has been determined at a resolution of 1.53 A. The crystals are orthorhombic, belonging to space group C222(1), with unit-cell parameters a = 49.03, b = 130.25, c = 157.29 A. The asymmetric unit contains two molecules. Phase information was derived from a multiwavelength anomalous dispersion (MAD) experiment conducted at three wavelengths using a selenomethionine-derivative crystal of HPSP. The structure was refined using CNS to a final crystallographic R value of 21.6% (R(free) = 23.4%). HPSP is a dimeric enzyme responsible for the third and final step of the l-serine biosynthesis pathway. It catalyses the Mg2+ dependent hydrolysis of l-phosphoserine. Recently, the structure of HPSP in complex with an inhibitor bound to the active site has been reported to be the open conformation of the enzyme. Here, the structure of HPSP is reported in the absence of substrate in the active site. Evidence is presented that HPSP in an uncomplexed form is in an even more open conformation than in the inhibitor complex. In this state, the enzyme is partially unfolded to allow the substrate to enter the active site. Binding of the substrate causes HPSP to shift to the closed conformation by stabilizing the partially unfolded region. In the present structure a Ca2+ ion is bound to the active site and an explanation is given why HPSP is not active when in the active site Mg2+ is replaced by a Ca2+ ion. PMID- 12777758 TI - The structure of endothiapepsin complexed with the gem-diol inhibitor PD-135,040 at 1.37 A. AB - The crystal structure of endothiapepsin complexed with the gem-diol inhibitor PD 135,040 has been anisotropically refined to a resolution of 1.37 A. The structure of this inhibitor complex is in agreement with previous structures of endothiapepsin gem-diol inhibitor complexes that have been used to develop proposed catalytic mechanisms. However, the increase in resolution over previous structures confirms the presence of a number of short hydrogen bonds within the active site that are likely to play an important role in the catalytic mechanism. The presence of low-barrier hydrogen bonds was indicated in a previous one dimensional H NMR spectrum. PMID- 12777759 TI - Analysis of the effect of microgravity on protein crystal quality: the case of a myoglobin triple mutant. AB - Crystals of the Met derivative of the sperm whale myoglobin triple mutant Mb-YQR [L(B10)Y, H(E7)Q and T(E10)R] were grown under microgravity conditions and on earth by vapour diffusion. A comparison of crystal quality after complete data collection and processing shows how microgravity-grown crystals diffract to better resolution and lead to considerably improved statistics for X-ray diffraction data compared with crystals grown on earth under the same conditions. The same set of experiments was reproduced on two different Spacelab missions (ISS 6A and ISS 8A) in 2001 and 2002. The structure of this mutant myoglobin, refined using data collected at ELETTRA (Trieste, Italy) from both kinds of crystals, shows that X-ray diffraction from microgravity-grown crystals leads to better defined electron-density maps as well as improved geometrical quality of the refined model. Improvement of the stereochemical parameters of a protein structure is fundamental to quantitative analysis of its function and dynamics and hence to thorough understanding of the molecular mechanisms of action. PMID- 12777760 TI - Impact of the physical and chemical environment on the molecular structure of Coprinus cinereus peroxidase. AB - The structure of the peroxidase from Coprinus cinereus (CiP) has been determined in three different space groups and crystalline environments. Two of these are of the recombinant glycosylated form (rCiP), which crystallized in space groups P2(1)2(1)2(1) and C2. The third crystal form was obtained from a variant of CiP in which the glycosylation sites have been removed (rCiPON). It crystallizes in space group P2(1) with beta approximately 90 degrees; the structure was determined from room-temperature data and low-temperature data obtained from twinned crystals. Two independent molecules of CiP related by non crystallographic symmetry are contained in the three crystal forms. The packing in the two structures of the glycosylated form of rCiP is closely related, but differs from the packing in the unglycosylated rCiPON. A database search based on small-molecule porphinato iron (III) complexes has been performed and related to observations of the spin states and coordination numbers of the iron ion. The room-temperature structures of CiP and one structure of the almost identical peroxidase from Arthromyces ramosus (ARP) have been used to identify 66 conserved water molecules and to assign a structural role to most of them. PMID- 12777761 TI - The structure of a mutant enzyme of Coprinus cinereus peroxidase provides an understanding of its increased thermostability. AB - Seven amino-acid substitutions introduced into the 343 amino-acid-long sequence of Coprinus cinereus peroxidase (CiP) led to a mutant enzyme (TS-rCiP) which is more stable than the native enzyme at higher temperature, pH and hydrogen peroxide concentrations. It is therefore more suitable for industrial applications. A structure determination was conducted on a deglycosylated but still active form of TS-rCiP based on X-ray diffraction data to 2.05 A resolution measured on a crystal cooled to 100 K and refined to R = 0.202 and R(free) = 0.249. The increased stability of the TS-rCiP enzyme can be understood from the structural changes of the TS-rCiP structure revealed by a comparative analysis with other known CiP structures. One of the more significant changes caused by three of the substitutions, I49S, V53A and T121A, is the conversion of a hydrophobic pocket into a hydrophilic pocket with associated changes in the water structure and the hydrogen-bonding interactions. The E239G substitution, which gives rise to increased thermostability at high pH, creates changes in the water structure and in the orientation of a phenylalanine (Phe236) in its vicinity. The three substitutions M166F, M242 and Y242F introduced to increase the oxidative stability do not introduce any structural changes. PMID- 12777762 TI - Structure of an RNA dodecamer containing a fragment from SRP domain IV of Escherichia coli. AB - The crystal structure of an RNA dodecamer, r(GCGUCAGGUC(Br)CG)/r(CGGAAGCAG(Br)CGC), containing a fragment from the signal recognition particle (SRP) RNA (domain IV) of Escherichia coli, has been determined at 1.7 A resolution with 21 666 independent reflections and an R(work) and R(free) of 20.1 and 22.5%, respectively. The structure exhibits a novel crystal packing pattern for RNA oligomer duplexes: one end of the duplex adopts the stacking interaction, while the other end adopts the abutting interaction in the minor groove. The symmetric loop of the SRP, r(CAGG)/r(AGCA), in the center of the dodecamer forms two different conformations of the A.C mismatch, a sheared G.G and a symmetrical G.A mismatch. These four mismatches present a unique surface for the abutting interaction. The involvement of the two A.C mismatches in the abutting interaction implies that these mismatches are the important sites for interaction with proteins. The conformation of the symmetric loop is greatly stabilized by hydrated metal ions, which display flexibility in adjusting their geometry and coordination in interaction with nucleic acids. Comparison with other crystal structures of fragments of 4.5S RNA indicates that the conformation of the symmetric loop is independent of the asymmetrical loop in domain IV. PMID- 12777763 TI - Solving the phase problem for carbohydrate-binding proteins using selenium derivatives of their ligands: a case study involving the bacterial F17-G adhesin. AB - The Escherichia coli adhesin F17-G is a carbohydrate-binding protein that allows the bacterium to attach to the intestinal epithelium of young ruminants. The structure of the 17 kDa lectin domain of F17-G was determined using the anomalous dispersion signal of a selenium-containing analogue of the monosaccharide ligand N-acetyl-d-glucosamine in which the anomeric oxygen was replaced by an Se atom. A three-wavelength MAD data set yielded good experimental phases to 2.6 A resolution. The structure was refined to 1.75 A resolution and was used to solve the structures of the ligand-free protein and the F17-G-N-acetyl-d-glucosamine complex. This selenium-carbohydrate phasing method could be of general use for determining the structures of carbohydrate-binding proteins. PMID- 12777764 TI - Low-resolution molecular replacement using a six-dimensional search. AB - A parallel-aware program MPI_FSEARCH has been implemented to perform molecular replacement using an exhaustive six-dimensional search. In particular, the program can be used to deal with diffraction data at very low resolution (d > 10 A) that would normally not be appropriate for other molecular-replacement programs such as AMoRe and CNS. Although an envelope constructed from a PDB (Protein Data Bank) file was tested in the present study, the program can be used to perform low-resolution molecular replacement with an envelope derived from various sources such as electron microscopy or small-angle solution X-ray scattering. PMID- 12777765 TI - Phasing on anomalous signal of sulfurs: what is the limit? AB - Recent years have witnessed significant advancements in X-ray data-acquisition techniques and phasing algorithms, which have made possible the successful use of a very small anomalous diffraction signal for the solution of crystal structures of macromolecules. Two crystal structures, a 44 kDa glucose isomerase containing nine sulfurs and a 33 kDa xylanase containing five sulfurs, have been solved from single-wavelength anomalous data using widely available methods and programs. These two enzymes contain less sulfur than most proteins in the bacterial or eukaryotic proteomes, providing a Bijvoet ratio of about 0.6%. For glucose isomerase the automatically interpretable electron-density maps could be obtained at high as well as low resolution. The S-SAD approach relies on the anomalous signal of sulfur naturally occurring in proteins and alleviates all need for sample derivatization. It may therefore be applicable to all protein crystals able to provide accurate diffraction data. PMID- 12777766 TI - Shotgun crystallization strategy for structural genomics: an optimized two-tiered crystallization screen against the Thermotoga maritima proteome. AB - As the field of structural genomics continues to grow and new technologies are developed, novel strategies are needed to efficiently crystallize large numbers of protein targets, thus increasing output, not just throughput [Chayen & Saridakis (2002). Acta Cryst. D58, 921-927]. One strategy, developed for the high throughput structure determination of the Thermotoga maritima proteome, is to quickly determine which proteins have a propensity for crystal formation followed by focused SeMet-incorporated protein crystallization attempts. This experimental effort has resulted in over 320 000 individual crystallization experiments. As such, it has provided one of the most extensive systematic data sets of commonly used crystallization conditions against a wide range of proteins to date. Analysis of this data shows that many of the original screening conditions are redundant, as all of the T. maritima proteins that crystallize readily could be identified using just 23% of the original conditions. It also shows that proteins that contain selenomethionine and are more extensively purified often crystallize in distinctly different conditions from those of their native less pure counterparts. Most importantly, it shows that the two-tiered strategy employed here is extremely successful for predicting which proteins will readily crystallize, as greater than 99% of the proteins identified as having a propensity to crystallize under non-optimal native conditions did so again as selenomethionine derivatives during the focused crystallization trials. This crystallization strategy can be adopted for both large-scale genomics programs and individual protein studies with multiple constructs and has the potential to significantly accelerate future crystallographic efforts. PMID- 12777767 TI - Purification, N-terminal sequencing, crystallization and preliminary structural determination of atratoxin-b, a short-chain alpha-neurotoxin from Naja atra venom. AB - Atratoxin-b, a short-chain alpha-neurotoxin purified from Naja atra (mainland Chinese cobra) venom using a three-step chromatography procedure, has an apparent molecular mass of 6950 Da with an alkaline pI value (>9.5) and consists of one single polypeptide chain as estimated by MALDI-TOF mass spectrometry and SDS PAGE. The protein is toxic to mice, with an in vitro LD(50) of about 0.18 mg kg( 1). Its N-terminal amino-acid sequence, LECHNQQSSQTPTIT, displays a very high homology to those of other alpha-neurotoxins. The overall three-dimensional structure of atratoxin-b is very similar to that of the homologous erabutoxin-a, as shown by the crystallographic molecular replacement and preliminary refinement results, with an R factor and R(free) of 27 and 29%, respectively. The microcrystal slowly grew to dimensions of approximate 0.1 x 0.1 x 0.15 mm over eight months using hanging-drop vapour-diffusion method. It gave a set of diffraction data to 1.56 A resolution using X-rays of wavelength 1.1516 A generated by the X-ray Diffraction and Scattering Station of beamline U7B at the National Synchrotron Radiation Laboratory (Hefei, China); this is the first example of the use of this beamline in protein crystallography. The crystals belong to the tetragonal space group P4(1)2(1)2, with unit-cell parameters a = 49.28, c = 44.80 A, corresponding to one molecule per asymmetric unit and a volume-to-mass ratio of 1.96 A(3) Da(-1). PMID- 12777768 TI - Crystallization and preliminary X-ray studies of the glutaredoxin from poplar in complex with glutathione. AB - A monocysteinic mutant of poplar glutaredoxin (C30S) has been overproduced and purified. The protein has been crystallized in complex with glutathione using the hanging-drop vapour-diffusion technique in the presence of PEG 4000 as a precipitating agent. A native data set was collected at 1.55 A resolution. The crystals belong to space group P2(1)2(1)2(1), with unit-cell parameters a = 45.7, b = 49.1, c = 104.8 A. Isomorphous crystals of a selenomethionine derivative were grown under the same conditions. Three data sets were collected at 1.73 A using the FIP synchrotron beamline at the ESRF. The positions of the Se atoms were determined and model rebuilding and refinement are in progress. PMID- 12777769 TI - Crystallization and preliminary X-ray analysis of Der f 2, a potent allergen derived from the house dust mite (Dermatophagoides farinae). AB - Although a number of allergens have been identified and isolated, the underlying molecular basis for the potent immune response is poorly understood. House dust mites (Dermatophagoides sp.) are ubiquitous contributors to atopy in developed countries. The rhinitis, dermatitis and asthma associated with allergic reactions to these arthropods are frequently caused by relatively small (125-129 amino acids) mite proteins of unknown biological function. Der f 2, a major allergen from the mite D. farinae, has been recombinantly expressed, characterized and crystallized. The crystals belong to the tetragonal space group I4(1)22, with unit-cell parameters a = b = 95.2, c = 103.3 A. An essentially complete (97.2%) data set has been collected to 2.4 A at a synchrotron source. Attempts to solve the crystal structure of Der f 2 by molecular replacement using the NMR coordinates for either Der f 2 or Der p 2 (the homologous protein from D. pteronyssinus) failed, but preliminary searches using the crystalline Der p 2 atomic coordinates appear to be promising. PMID- 12777770 TI - Crystallization and preliminary X-ray diffraction analysis of Langat virus envelope protein domain III. AB - The putative receptor-binding domain (domain III) of the flavivirus Langat envelope glycoprotein has been crystallized using the hanging-drop vapor diffusion method at 277 K. Two distinct crystal morphologies were observed to grow under the same conditions. The crystal forms both belong to a trigonal space group, P3(1)21 or P3(2)21, with unit-cell parameters a = 80.93, c = 132.1 A and a = 104.8, c = 219.5 A for forms I and II, respectively. Complete data sets to 2.9 and 3.35 A, respectively, have been collected at 100 K with Cu Kalpha X-rays from a rotating-anode generator. PMID- 12777771 TI - Crystallization and preliminary X-ray diffraction analysis of Jun a 1, the major allergen isolated from pollen of the mountain cedar Juniperus ashei. AB - Jun a 1, the major allergen of pollen from the mountain cedar Juniperus ashei, has been crystallized using the hanging-drop vapor-diffusion method at 277 K. The crystals are monoclinic, space group P2(1), with unit-cell parameters a = 53.38, b = 113.48, c = 72.44 A, beta = 96.36 degrees and four molecules in the unit cell. A complete 2.5 A data set has been collected at 100 K with X-rays from a Cu Kalpha rotating-anode generator. PMID- 12777772 TI - Crystallization and preliminary X-ray analysis of Weissella viridescens FemX UDP MurNAc-pentapeptide:l-alanine ligase. AB - Synthesis of the cell-wall peptidoglycan of firmicutes involves a unique family of peptide-bond-forming enzymes that use amino-acyl-tRNAs as substrates and are referred to as Fem proteins as they are factors essential for methicillin resistance in Staphylococcus aureus. The FemX UDP-MurNAc-pentapeptide:l-alanine ligase of Weissella viridescens was overexpressed, purified and crystallized. Native data were collected to 1.7 A resolution. The crystals belong to space group P2(1), with unit-cell parameters a = 42.03, b = 99.92, c = 45.84 A, beta = 116.02 degrees. The asymmetric unit contains one molecule. A selenium-derivative data set has been collected to 2.1 A resolution at the peak wavelength of the selenium absorption edge. Six strong selenium positions were visible in the anomalous Patterson map. Three additional weaker Se atoms have been identified by anomalous Fourier synthesis. PMID- 12777773 TI - Expression, purification, crystallization and preliminary X-ray characterization of the class B acid phosphatase (AphA) from Escherichia coli. AB - The class B non-specific acid phosphatase AphA from Escherichia coli has been expressed in E. coli and purified following a new protocol. ESI mass spectroscopy shows that the purified enzyme solution contains two polypeptides with molecular weights differing by 185 Da corresponding to two different cleavage sites of the signal peptide from the AphA E. coli precursor. Despite the solution heterogeneity, X-ray quality crystals have been obtained. However, the crystals have a tendency to give polymorphs and to lose long-range order with time while maintaining an intact crystal habit. Crystals have been grown in space groups I222 and C2 with three different unit cells and different asymmetric unit contents. Diffraction data to 1.6 A resolution have been collected with synchrotron radiation at ESRF and DESY. PMID- 12777774 TI - Crystallization of Hfq protein: a bacterial gene-expression regulator. AB - Hfq protein from Escherichia coli (EcoHfq) has been overproduced in E. coli, purified to homogeneity and crystallized using the hanging-drop vapour-diffusion technique. Crystallization conditions for EcoHfq were found which yielded X-ray quality crystals. Crystals of EcoHfq and of Cd-, Hg- and Se-containing derivatives grew in two months, with unit-cell parameters a = b = 127.41, c = 170.36 A. The crystals belong to space group I4 and diffract to 2.1 A resolution. Two hexamers are predicted per asymmetric unit. PMID- 12777775 TI - Crystallization and preliminary X-ray analysis of Escherichia coli p20, a novel thiol peroxidase. AB - Escherichia coli p20 is a thioredoxin-dependent thiol peroxidase. This protein represents a novel group of antioxidant enzymes that are widely expressed in various pathogenic bacteria and show distant yet significant sequence homology with peroxiredoxins. E. coli p20, overexpressed in E. coli, was crystallized with PEG 4000 and 2-propanol as precipitants using the hanging-drop vapour-diffusion method. Diffraction data were collected to 2.2 A resolution using synchrotron radiation. The crystals belong to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 38.97, b = 58.97, c = 127.59 A. The asymmetric unit contains two p20 molecules, with a corresponding V(M) of 2.06 A(3) Da(-1) and a solvent content of 40.4%. PMID- 12777776 TI - Structural studies of the transpeptidase domain of PBP1a from Streptococcus pneumoniae. AB - The synthesis of the bacterial cell wall requires enzymes which are localized both in the cytoplasm and in the periplasm. Penicillin-binding proteins (PBPs) catalyze the last, crucial steps in peptidoglycan biosynthesis and several of them are essential for bacterial survival. High-molecular-mass PBPs can be bifunctional (class A) or monofunctional (class B) and to date no structural information on any class A PBP is available. To initiate the determination of the three-dimensional structure of a class A PBP, crystals of the transpeptidase domain of PBP1a from Streptococcus pneumoniae were prepared by limited proteolysis of the full-length molecule and purification by anion-exchange chromatography and gel filtration. The samples crystallize in space group C222(1), contain one molecule per asymmetric unit and diffract X-rays to 2.7 A. Selenomethionine-labelled crystals have been prepared and structure solution is under way. PMID- 12777777 TI - Crystallization and preliminary X-ray diffraction studies of the eukaryotic iron superoxide dismutase (FeSOD) from Vigna unguiculata. AB - Eukaryotic iron superoxide dismutases (FeSODs) are homodimeric proteins that constitute a fundamental protection against free radicals, which can damage essential cellular mechanisms. The protein was cloned and overexpressed in Escherichia coli with an N-terminal His tag. Crystallization experiments of the protein resulted, after several refined screenings, in crystals suitable for X ray diffraction analysis. The crystals belong to the monoclinic space group C2, with unit-cell parameters a = 82.54, b = 48.41, c = 64.28 A, alpha = gamma = 90, beta = 119.66 degrees, and contain one molecule per asymmetric unit. At cryogenic temperatures, the crystals diffracted to a resolution limit of 1.80 A using synchrotron radiation at the European Synchrotron Radiation Facility (ESRF). PMID- 12777778 TI - Purification, crystallization and preliminary X-ray crystallographic studies on acetolactate decarboxylase. AB - Acetolactate decarboxylase has the unique ability to decarboxylate both enantiomers of acetolactate to give a single enantiomer of the decarboxylation product, (R)-acetoin. A gene coding for alpha-acetolactate decarboxylase from Bacillus brevis (ATCC 11031) was cloned and overexpressed in B. subtilis. The enzyme was purified in two steps to homogeneity prior to crystallization. Three different diffraction-quality crystal forms were obtained by the hanging-drop vapour-diffusion method using a number of screening conditions. The best crystal form is suitable for structural studies and was grown from solutions containing 20% PEG 2000 MME, 10 mM cadmium chloride and 0.1 M Tris-HCl pH 7.0. They grew to a maximum dimension of approximately 0.4 mm and belong to the trigonal space group P3(1,2)21, with unit-cell parameters a = 47.0, c = 198.9 A. A complete data set was collected to 2 A from a single native crystal using synchrotron radiation. PMID- 12777779 TI - Crystallization and preliminary X-ray analysis of the E. coli hypothetical protein TdcF. AB - Crystals of the hypothetical protein TdcF (subunit MW = 14 007) from Escherichia coli were grown by vapour diffusion. The protein crystallizes in space group P2(1)2(1)2, with unit-cell parameters a = 72.67, b = 86.22, c = 62.62 A. Native data to a resolution of 2.35 A were collected from a single crystal at 100 K on a rotating-anode X-ray generator. Preliminary analysis of these data indicated that the asymmetric unit corresponded to a trimer, which was supported by a convincing molecular-replacement solution using the YjgF trimer as the probe structure. PMID- 12777780 TI - Crystallization and preliminary X-ray characterization of a novel calcium-binding protein AtCBL2 from Arabidopsis thaliana. AB - A new family of calcineurin B-like calcium-binding proteins has recently been identified in Arabidopsis thaliana. AtCBL2, a member of this family, has been crystallized in the presence of calcium ions using polyethylene glycol as a precipitant at 293 K. The crystals belong to space group C222(1), with unit-cell parameters a = 83.9, b = 118.1, c = 49.1 A. The asymmetric unit contains one molecule, with a V(M) of 2.36 A(3) Da(-1) and a solvent content of 48%. Native diffraction data to 2.1 A resolution have been collected using synchrotron radiation at SPring-8. PMID- 12777781 TI - Expression and preliminary crystallographic studies of R1E, the large subunit of ribonucleotide reductase from Salmonella typhimurium. AB - The nrdE gene product R1E, the large subunit of the class 1b Salmonella typhimurium ribonucleotide reductase, has been overexpressed, purified and crystallized. Initially, the protein crystallized in two orthorhombic space groups, C222(1) and P2(1)2(1)2, using tartrate and PEG 6000 as precipitants, respectively. Better diffracting crystals belonging to the tetrahedral space group P4(3)2(1)2 were obtained using sodium malonate as precipitant. The P4(3)2(1)2 crystals could only be obtained after seeding from a drop containing C222(1) crystals grown in sodium tartrate. Thus, streak-seeding resulted in crystals of a supergroup to C222(1). Data to 2.8 A resolution have been collected on the P4(3)2(1)2 crystals which contained one R1E subunit in the asymmetric unit. PMID- 12777782 TI - Identification, cDNA cloning, expression, crystallization and preliminary X-ray analysis of an exceptionally halotolerant carbonic anhydrase from Dunaliella salina. AB - An extracellular alpha-type carbonic anhydrase (dCAII) from the salt-tolerant alga Dunaliella salina differs from its mesophilic counterparts in remaining active from zero to multimolar salt concentrations. To gain insight into the outstanding salt tolerance of dCAII, the enzyme was functionally overexpressed in Escherichia coli, purified by affinity chromatography and crystallized by the hanging-drop method. The crystals belonged to space group P2(1), with unit-cell parameters a = 47.0, b = 119.9, c = 58.5 A, beta = 94.2 degrees. Data from a single crystal were collected to 2.4 A resolution under cryogenic conditions (120 K) using an R-AXIS IV(++) detector mounted on a Rigaku RU-H3R rotating-anode generator. The asymmetric unit contains two molecules of the protein, which corresponds to V(M) = 2.65 A(3) Da(-1) and a solvent content of 52.7%. PMID- 12777783 TI - Cloning, preliminary characterization and crystallization of nucleoside hydrolases from Caenorhabditis elegans and Campylobacter jejuni. AB - The nucleoside hydrolases (NHs) are a family of nucleoside-modifying enzymes. They play an important role in the purine-salvage pathway of many pathogenic organisms which are unable to synthesize purines de novo. Although well characterized in protozoan parasites, their precise function and mechanism remain unclear in other species. For the first time, NHs from Caenorhabditis elegans and Campylobacter jejuni, which are representatives of mesozoa and bacteria, respectively, have been cloned and purified. Steady-state kinetics indicate a different substrate-specificity profile to previously described hydrolases. Native diffraction data sets were collected from crystals of NH from each organism. The hexagonal crystals (space group P6(2)22 or P6(4)22) of NH from C. elegans diffracted to a resolution of 2.8 A, while the data set from the orthorhombic crystals (space group I222 or I2(1)2(1)2(1)) of NH from C. jejuni could be processed to 1.7 A resolution. The unit-cell parameters were a = b = 102.23, c = 117.27 A in the former case and a = 101.13, b = 100.13, c = 81.37 A in the latter. PMID- 12777784 TI - Crystallization of the Bacillus subtilis SPP1 bacteriophage helicase loader protein G39P. AB - The essential helicase loader protein G39P encoded by Bacillus subtilis SPP1 phage has been overproduced in Escherichia coli and purified. The wild-type protein has been crystallized by the hanging-drop vapour-diffusion method in a primitive hexagonal space group, probably P6(1)22/P6(5)22, but the crystals diffract to only 3.4 A and are poorly reproducible. Mass-spectrometric analysis has revealed marked proteolytic cleavage from the C-terminus and the presence of a major species corresponding to deletion of the 14 C-terminal residues. Thus, a new variant of the protein (G39P112) has been engineered that corresponds to a 14 residue C-terminal truncation. The G39P112 variant has also been crystallized but now in a primitive orthorhombic form, probably P2(1)2(1)2 or P2(1)2(1)2(1), with unit-cell parameters a = 85.6, b = 89.7, c = 47.6 A, with diffraction to 2.4 A on a synchrotron source and with greatly improved reproducibility. Calculation of V(M) values for this G39P112 variant suggests the presence of three monomers in the asymmetric unit, corresponding to a solvent content of about 47%. A selenomethionine-incorporated form of the protein has been produced and a full three-wavelength MAD data collection undertaken. PMID- 12777785 TI - Crystallization and X-ray diffraction data analysis of leukotriene A4 hydrolase from Saccharomyces cerevisiae. AB - The Saccharomyces cerevisiae leukotriene A4 (LTA4) hydrolase (scLTA4 hydrolase) has been crystallized in order to study the two activities of LTA4 hydrolase in an evolutionary perspective. Single well diffracting crystals are obtained after switching from the hanging-drop method to liquid-liquid diffusion in capillaries using PEG 8000 as precipitant. These crystals belong to space group P2(1)2(1)2(1), with unit-cell parameters a = 70.8, b = 98.1, c = 99.2 A. Intensity data to 2.3 A resolution were collected from a native scLTA4 hydrolase crystal using synchrotron radiation. A molecular-replacement solution was obtained using the human LTA4 hydrolase structure and the program BEAST. PMID- 12777786 TI - Cloning, expression and preliminary crystallographic studies of the potential drug target purine nucleoside phosphorylase from Schistosoma mansoni. AB - The parasite Schistosoma mansoni, unlike its mammalian hosts, lacks the de novo pathway for purine biosynthesis and depends on salvage pathways for its purine requirements. The gene encoding one enzyme of this pathway, purine nucleoside phosphorylase from S. mansoni (SmPNP) was identified, fully sequenced and cloned into the bacterial expression vector pMAL c2G to produce a protein in fusion with maltose-binding protein. The recombinant fusion protein was expressed at high levels and was purified in a single step by amylose resin affinity chromatography. After factor Xa cleavage, SmPNP was purified using a cation exchange column and crystallized by hanging-drop vapour diffusion using polyethylene glycol 1500 as precipitant in the presence of 20% glycerol in acetate buffer. The use of the non-detergent sulfobetaine 195 (NDSB 195) as an additive had a marked effect on the size of the resulting crystals. Two data sets were obtained, one from a crystal grown in the absence of NDSB 195 and one from a crystal grown in its presence. The crystals are isomorphous and belong to the space group P2(1)2(1)2(1). It is intended to use the structures in the discovery and development of specific inhibitors of SmPNP. PMID- 12777787 TI - Crystallization of butyrate kinase 2 from Thermotoga maritima mediated by vapor diffusion of acetic acid. AB - The sitting-drop method of crystallization uses the evaporation of water to increase the concentration of the protein and precipitant in the drop. The presence of other volatile components, such as acetic acid, can have a marked impact on crystallization. A member of the ASKHA (acetate and sugar kinases/Hsc70/actin) superfamily of proteins, isobutyrate kinase (Buk2) from Thermotoga maritima, was expressed in Escherichia coli with six histidine residues added to the C-terminus. The purified protein was crystallized in a sitting drop with a well solution consisting of 1.7-3.0 M sodium formate, with the pH of the well solution alone adjusted to 4.5 with acetic acid. Diffraction data collected at 100 K show that the crystals diffract to 3.1 A and belong to space group I422, with unit-cell parameters a = b = 198.12, c = 58.93 A. Both the crystal form and the results of dynamic light-scattering studies suggest that Buk2 is an octomer, the first to be identified in the ASKHA superfamily. PMID- 12777788 TI - Production, crystallization and preliminary crystallographic study of the major cat allergen Fel d 1. AB - The domestic cat (Felis domesticus) is an important cause of allergic disease worldwide. The major cat allergen 1 (Fel d 1) has been expressed in Escherichia coli, purified and refolded in a soluble form. Crystals of Fel d 1 were obtained in 13% 2-methyl-2,4-pentanediol, 0.1 M sodium acetate pH 4.8. The Fel d 1 crystals belong to space group P2(1), with unit-cell parameters a = 43.3, b = 51.5, c = 67.7 A, and diffract to 1.9 A resolution. PMID- 12777789 TI - Purification, nanocrystallization and preliminary X-ray analysis of a C-terminal part of tropomodulin protein 1, isoform A, from Caenorhabditis elegans. AB - The C-terminal part of tropomodulin protein 1, isoform A, from Caenorhabditis elegans was expressed in Escherichia coli and purified to homogeneity. Optimized from the initial nanoscreen, crystals grew to dimensions of 0.25 x 0.15 x 0.15 mm at 277 K using 28.0%(v/v) PEG 400 as the precipitant by the hanging-drop vapor diffusion technique. A data set of 94.9% completeness was collected to a resolution of 1.98 A at 100 K using a synchrotron X-ray source (SER-CAT). The crystals belong to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 31.7, b = 50.6, c = 107.1 A, and contained one molecule per asymmetric unit. PMID- 12777790 TI - Purification, crystallization and preliminary X-ray studies of GMP reductase 2 from human. AB - GMP reductase 2 from human has been expressed in Escherichia coli, purified and crystallized. The crystals belong to space group P3(2)21, with unit-cell parameters a = b = 110.6, c = 209.8 A, alpha = beta = 90, gamma = 120 degrees. Diffraction data were collected to 3.0 A with a completeness of 100% (100% for the last shell), an R(merge) value of 0.089 (0.189) and an I/sigma(I) value of 7.3 (3.2). PMID- 12777791 TI - The structure of human phosphoglucose isomerase complexed with a transition-state analogue. AB - Phosphoglucose isomerase (PGI) is a workhorse enzyme of carbohydrate metabolism that interconverts glucose 6-phosphate and fructose 6-phosphate. Outside the cell, however, the protein appears to function as a cytokine. A crystal structure of human PGI bound with 5-phosphoarabinonate, a strong inhibitor that mimics the cis-enediol(ate) intermediate of the reaction, has been determined at 2.5 A resolution. The structure helps to confirm the assignment of Glu357 as the base catalyst in the isomerase reaction. PMID- 12777792 TI - Compatibility of detergents with the microbatch-under-oil crystallization method. AB - Detergents are required to solubilize integral membrane proteins and are common components of the solutions used to crystallize these molecules. It has been unclear whether these detergents are completely compatible with the oils used in the microbatch-under-oil crystallization technique, because they might conceivably be lost from solution by partitioning into the oil phase. The partitioning of the detergents n-octyl-beta-d-glucopyranoside and Fos-Choline-12 into two different oils used for microbatch crystallization experiments has been examined. It was found that vigorous mixing and prolonged incubation of the aqueous detergent solutions with the oils leads to small losses of detergent (approximately 5% of the total detergent mass); however, gentle mixing that is more typical of the mixing encountered in a crystallization experiments leads to negligible loss of detergent. PMID- 12777793 TI - The architecture of the GroEL-GroES-(ADP)(7) chaperonin complex. I. Heptagrammal molecular forms. AB - Molecular forms are considered with vertices that have integral coordinates (the indices) with respect to a symmetry-adapted basis and which are left invariant by a point group of crystallographic scale-rotations (represented in this basis by invertible integral matrices). The composite form enclosing the chaperonin complex GroEL-GroES-(ADP)(7) is derived and decomposed into heptagrammal forms. These are generalizations of the two-dimensional forms based on sevenfold star polygons. In the chaperonin complex, nine such heptagrammal molecular forms are found: three for each ring (trans and cis) of GroEL and three for GroES. These forms correspond to a splitting of the monomer into adjacent segments. The change in the folding of the chains in the cis ring of GroEL arising from binding to GroES leaves the chain segmentation invariant. PMID- 12777794 TI - The architecture of the GroEL-GroES-(ADP)(7) chaperonin complex. II. Heptagrammal characterization of the folding. AB - The heptagrammal forms derived in part I [Janner (2003a). Acta Cryst. D59, 783 794] enclose chain segments of symmetry-related monomers in the GroEL-GroES (ADP)(7) chaperonin complex. A chain reaching the boundary of a given form either ends, proceeds to a neighbouring form or has to fold. C(alpha) atoms corresponding to these folding points are identified in each of the nine forms of the chaperonin and are approximated by ideal positions having integral coordinates (the indices) with respect to a symmetry-adapted basis. Mutual structural relations between the indexed positions are derived in terms of integral scale-rotations (similar to those that leave the form invariant). The magnesium ions at the binding sites of the nucleotides ADP and ATP are shown to be symmetry-related to these folding points. The change in folding (polymorphism) observed in the cis ring of GroEL arising from binding to GroES is discussed. In particular, the form segmentation is conserved in the polymorphic transition. The geometric and algebraic restrictions imposed on the indexed positions and on their structural relations by the integrality condition are presented in an appendix. PMID- 12777795 TI - Recursive direct phasing with reference-beam diffraction. AB - The reference-beam diffraction technique provides a practical way to measure a large number of triplet phases in a standard oscillating-crystal diffraction experiment for protein crystals. The triplet-phase data set from such reference beam measurements contains a unique phase-occurrence pattern that leads to a new recursive phasing algorithm for the individual structure-factor phases. Application of the new algorithm is demonstrated for tetragonal lysozyme using 7360 triplet phases measured in a reference-beam experiment with a median phase discrepancy of 45 degrees. An electron-density map obtained using this phasing algorithm and the measured triplet phases shows good agreement with the known protein structure. PMID- 12777796 TI - Structure of DNA helicase RepA in complex with sulfate at 1.95 A resolution implicates structural changes to an "open" form. AB - The structure of a new crystal form (space group C2), grown at pH 8.0 and diffracting to 1.95 A resolution, of the replicative homo-hexameric DNA helicase RepA encoded by plasmid RSF1010 is reported. In contrast to previous crystals grown at pH 6.0 in space group P2(1) (Niedenzu et al., 2001), only one half (a trimer) of the RepA hexamer occupies the asymmetric unit of the space-group C2 crystals. The new crystal packing explains the pH-dependent hexamer-hexamer association mechanism of RepA. The C-terminus (264)VLERQRKSKGVPRGEA(279), which could not be modelled in the previous structure, is clearly defined in the present electron density except for the last four amino acids. Sulfate anions occupy the six ATPase active sites of RepA at positions where the product phosphates are supposed to bind. Binding of sulfate anions induces conformational changes both at the ATPase active sites and throughout the whole molecular structure. In agreement with electron microscopy, the above studies implicate structural changes to an "open" form that may occur upon binding and hydrolysis of nucleotide 5'-triphosphates and could be essential for DNA duplex-unwinding activity. PMID- 12777797 TI - An evaluation of automated model-building procedures for protein crystallography. AB - The computer programs ARP/wARP, MAID and RESOLVE are designed to build protein structures into experimentally phased electron-density maps without any user intervention, requiring only diffraction data and sequence information. However, the MAID and RESOLVE systems, which seek to extend the range of automated model building to approximately 3 A resolution, have yet to receive significant testing outside the small numbers of data sets used in their development. Since these two systems employ a large number of scoring functions and decision-making heuristics, additional tests are required to establish their usefulness to the crystallographic community. To independently evaluate these programs, their performance was tested using a database containing 41 experimentally phased maps between 1.3 and 2.9 A resolution from a diverse set of protein structures. At resolutions higher than 2.3 A the most successful program was ARP/wARP 6.0, which accurately built an average of 90% of the main chain. This system builds somewhat larger fractions of the model than the previous version ARP/wARP 5.1, which accurately built an average of 87% of the main chain. Although not specifically designed for model building into high-resolution maps, MAID and RESOLVE were also quite successful in this resolution regime, typically building approximately 80% of the main chain. At 2.3-2.7 A resolution the MAID and RESOLVE programs automatically built approximately 75% of the main-chain atoms in the protein structures used in these tests, which would significantly accelerate the model building process. Data sets at lower resolution proved more problematic for these programs, although many of the secondary-structure elements were correctly identified and fitted. PMID- 12777798 TI - Structural characterization and comparison of RGD cell-adhesion recognition sites engineered into streptavidin. AB - The RGD (arginine-glycine-aspartic acid) sequence is found in several important extracellular matrix proteins and serves as an adhesion ligand for members of the integrin family of cell-surface receptors. This sequence and flanking residues from fibronectin or osteopontin have been engineered into an accessible surface loop of streptavidin to create two new streptavidin variants (FN-SA or OPN-SA, respectively) that bind cells through the alpha(v)beta(3) and/or alpha(5)beta(1) integrin receptors. Their crystal structures confirm the design and construction of the mutants and provide evidence about the conformational dynamics of the RGD loops. The loops in the isomorphous crystal structures are involved in crystal packing interactions and this stabilizes their structures. Even so, the loop in OPN-SA is slightly disordered and two of the residues are not seen in difference electron-density maps. Comparison with other experimentally determined structures of RGD loops in cell-adhesion molecules shows that these loops occupy a large subset of conformational space. This is consistent with the view that RGD loops, at least those involved in cell adhesion, sample a number of structures dynamically, a few of which display high affinity for appropriate receptors. PMID- 12777799 TI - The structure of glyceraldehyde 3-phosphate dehydrogenase from Alcaligenes xylosoxidans at 1.7 A resolution. AB - The enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from the Gram negative denitrifying bacterial species Alcaligenes xylosoxidans was purified and crystallized as a contaminant protein during purification of nitrous oxide reductase. This is the first structure of a GAPDH from a denitrifying species. The crystal structure was solved at 1.7 A resolution by molecular replacement using the structure of GAPDH from Bacillus stearothermophilus as a starting model. The quality of the structure enabled the amino-acid sequence of the A. xylosoxidans GAPDH to be assigned. The structure is that of the apo-enzyme, lacking the NAD+ cofactor and with the active-site residue Cys154 oxidized. The global structure of the enzyme has a homotetrameric quaternary structure similar to that observed for its bacterial and eukaryotic counterparts. The essential role of Cys154 in the enzyme activity has been confirmed. In monomer O two half occupancy sulfate ions were found at the active site, which are analogous to the substrate and the "attacking" phosphate seen in B. stearothermophilus. One half occupancy sulfate ion is also located in the substrate-binding site of monomer P. PMID- 12777800 TI - The properties of (2Fo - Fc) and (Fo - Fc) electron-density maps at medium-to high resolutions. AB - This paper reports on the efficacy of (F(o) - F(c)) versus (2F(o) - F(c)) electron-density maps at 3.2 A resolution. Firstly, a study is reported of a simple truncation at 2.3 and 3.2 A of the 1.6 A resolution crystal structure of concanavalin A at room temperature [Emmerich et al. (1994), Acta Cryst. D50, 749 756] with 149 known bound water molecules. Secondly, the concanavalin A 1.6 A resolution model was re-refined but with the data truncated to 3.2 A. In a similar evaluation, these procedures were repeated for the apocrustacyanin A1 cryotemperature 1.4 A resolution model [Cianci et al. (2001), Acta Cryst. D57, 1219-1229]. Maps at 1.4, 2.3 and 3.2 A resolutions were first generated and the structure was then re-refined at 3.2 A and additionally at 2.3 A resolution. The results on concanavalin A show that the number of bound water molecules that are resolved decreases by two thirds from 1.6 to 3.2 A, but that key structural waters, for example at the transition metal and the calcium ion, are still resolved in the (F(o) - F(c)) map but not in the (2F(o) - F(c)) map. For apocrustacyanin A1, the results with these two difference maps were less clear cut. Two key structural bound waters (w93 and w105) were selected that had been previously identified in beta-crustacyanin [Cianci et al. (2002), Proc. Natl Acad. Sci. USA, 99, 9795-9800] in protein-carotenoid interactions. The behaviour of w93 is similar to that of concanavalin A key waters, but that of w105 is not. These behaviours were therefore explored in finer resolution increments, namely 2.9, 2.7 and 2.5 A. Finally, further tests on "real" data sets for peanut lectin and concanavalin A at medium resolution confirm these map properties, namely that an (F(o) - F(c)) difference electron-density map is more effective than a (2F(o) F(c)) map in showing bound water structure at lower resolutions ( approximately 3.2 A). This result is important since a growing number of protein crystal structure studies are concerned with multi-macromolecular complexes and are at such resolutions. Details of the bound solvent can still be revealed at 3.2 A via the (F(o) - F(c)) map calculation. The physical basis of the limitation of the (2F(o) - F(c)) map presumably lies in the series-termination error effect on such a map involving the first negative ripple from the protein atom to which a bound water oxygen is hydrogen bonded, sufficiently cancelling its peak. In addition, re-refinements at 3.2 A show distances that can agree with known values but B values that do not agree with known values. PMID- 12777801 TI - Connectivity-based ab initio phasing at different solvent levels. AB - The connectivity-based phasing method has been applied independently to three neutron diffraction data sets obtained from the same crystal of tRNA(Asp) aspartyl-tRNA synthetase complex but at different concentrations of D(2)O/H(2)O, thus masking different components of the crystal. The obtained low-resolution images correlate perfectly with the solvent level. PMID- 12777802 TI - Structure of the human S100A12-copper complex: implications for host-parasite defence. AB - S100A12 is a member of the S100 family of EF-hand calcium-modulated proteins. Together with S100A8 and S100A9, it belongs to the calgranulin subfamily, i.e. it is mainly expressed in granulocytes, although there is an increasing body of evidence of expression in keratinocytes and psoriatic lesions. As well as being linked to inflammation, allergy and neuritogenesis, S100A12 is involved in host parasite response, as are the other two calgranulins. Recent data suggest that the function of the S100-family proteins is modulated not only by calcium, but also by other metals such as zinc and copper. Previously, the structure of human S100A12 in low-calcium and high-calcium structural forms, crystallized in space groups R3 and P2(1), respectively, has been reported. Here, the structure of S100A12 in complex with copper (space group P2(1)2(1)2; unit-cell parameters a = 70.6, b = 119.0, c = 90.2 A) refined at 2.19 A resolution is reported. Comparison of anomalous difference electron-density maps calculated with data collected with radiation of wavelengths 1.37 and 1.65 A shows that each monomer binds a single copper ion. The copper binds at an equivalent site to that at which another S100 protein, S100A7, binds zinc. The results suggest that copper binding may be essential for the functional role of S100A12 and probably the other calgranulins in the early immune response. PMID- 12777803 TI - SAD manganese in two crystal forms of glucose isomerase. AB - Glucose isomerase from Streptomyces rubiginosus was crystallized in two forms: I222, with one molecule of 44 kDa in the asymmetric unit, and P2(1)2(1)2, with two unique molecules. The I222 structure is known, but the P2(1)2(1)2 form has not been solved before. X-ray diffraction data for the P2(1)2(1)2 form were collected at a wavelength of 1.54 A and data for the I222 form were collected at three different wavelengths: 1.34, 1.07 and 0.98 A. The amount of anomalous signal from one Mn and eight S atoms in these data sets varies from 1.24% to as low as 0.56%. The dual-space direct-methods program SHELXD, run against the Bijvoet differences, gave a clear solution of all anomalous scatterers for all data sets. The Mn positions only were used for SAD phasing of all four data sets. The electron-density map after density modification, resulting from the phasing of a single-wavelength data set and based purely on the anomalous deltaf" contribution, was clearly interpretable; an almost complete model of the protein was built by wARP without human intervention in all four cases. As far as is known, this is the first time that an anomalous signal as low as 0.6% has successfully been used to determine the structure of a macromolecule. PMID- 12777804 TI - Structure of a constitutively activated RhoA mutant (Q63L) at 1.55 A resolution. AB - Mutants of the small G protein RhoA that are deficient in GTPase activity and thereby exhibit constitutive molecular signaling activity are commonly used to discover its cellular functions. In particular, two such mutants, Gly14-->Val (G14V) and Gln63-->Leu (Q63L), are often used interchangeably for such studies. However, while their in vitro rates of GTP hydrolysis are very similar, differences are observed in their other functional properties. The structure of G14V-RhoA is known; in order to assess whether structural variations are responsible for functional differences, the crystal structure of a Q63L-RhoA bound to the GTP-analog 5'-guanylylimidodiphosphate (GMPPNP) was determined at 1.5 A resolution. Overall, the structure is very similar to that of G14V-RhoA, but the significantly higher resolution data permit an improved basis for structural analysis and comparison. The data support the notion that differences observed between the mutants in vivo are likely to arise from altered affinities for RhoGDI and not from direct structural differences. PMID- 12777805 TI - Characterization of four orthologs of stringent starvation protein A. AB - Orthologous proteins can be beneficial for X-ray crystallographic studies when a protein from an organism of choice fails to crystallize or the crystals are not suitable for structure determination. Their amino-acid sequences should be similar enough that they will share the same fold, but different enough so that they may crystallize under alternative conditions and diffract to higher resolution. This multi-species approach was employed to obtain diffraction quality crystals of the RNA polymerase (RNAP) associated stringent starvation protein A (SspA). Although Escherichia coli SspA could be crystallized, the crystals failed to diffract well enough for structure determination. Therefore, SspA proteins from Yersinia pestis, Vibrio cholerae and Pseudomonas aeruginosa were cloned, expressed, purified and subjected to crystallization trials. The V. cholerae SspA protein failed to crystallize under any conditions tested and the P. aeruginosa SspA protein did not form crystals suitable for data collection. On the other hand, Y. pestis SspA crystallized readily and the crystals diffracted to 2.0 A. PMID- 12777806 TI - Phasing power at the K absorption edge of organic arsenic. AB - Single/multiple-wavelength anomalous dispersion (SAD/MAD) experiments were performed on a crystal of an organic arsenic derivative of hen egg-white lysozyme. A para-arsanilate compound used as a crystallizing reagent was incorporated into the ordered solvent region of the lysozyme molecule. Diffraction data were collected to high resolution (/=16 years of age). Twelve patients (6.4%) received two and three retransplants (re-LT). Split grafting was performed 19 times (19 of 202 grafts, 9.4%). Risk factors included United Network of Organ Sharing status I (n=30, 16%), previous right upper abdominal surgery (n=32, 17.1%), caudate lobe encirclement of IVC (n=65, 32.2%), IVC (n=24, 11.9%), and splanchnic venous modification (n=58, 30.9%), transjugular intrahepatic portosystemic stent shunt (n=34, 16.8%), giant (>5 kg) liver tumor (n=6, 3%), septic necrosis of the caudate lobe (n=1, 0.5%), and previous cavocaval (n=13, 6.4%) or classical LT (n=5, 2.5%). RESULTS: IVC preservation, avoidance of IVC cross clamping and of VVB use were possible in 98.9%, 93%, and 99.5% of 183 primary LT and in 89.5%, 84.2%, and 89.5% of 19 re LT. Temporary portocaval shunting was never applied. Perioperative mortality was 1.2%. There was no allotransfusion in 73 (36%) grafts and 45 (22%) patients were immediately extubated. Permanent hepatic vein and caval problems were encountered in three (1.5%) grafts. One patient needed stent placement to treat IVC stenosis. Actual 3- and 12-month patient survival for whole, re-LT, and right-lobe split LT groups were 94.7%, 94.1%, 94.7%, 88.2%, 94.1%, and 89%. Three-month graft survival rates for these groups were 92.6%, 94.7%, and 84.2%. CONCLUSIONS: LT with IVC preservation and without VVB use and portocaval shunting is possible in nearly all primary transplants and in the majority of re-LT. PMID- 12777867 TI - Improvement of inflammatory bowel disease after allogeneic stem-cell transplantation. AB - Because causes of inflammatory bowel disease (IBD) remain obscure and a curative therapy is still lacking, the influence of stem-cell transplantation (SCT) on IBD is of major interest. We retrospectively analyzed the course of seven patients with Crohn's disease and four patients with idiopathic ulcerative colitis who underwent allogeneic SCT between July 1994 and August 2002 for acute and chronic myeloid leukemia and myelodysplastic syndrome. After a median follow-up of 34 months posttransplantation, 10 patients are alive. None of the patients showed IBD activity after SCT, except one patient with mild persistent symptoms of Crohn's disease early after transplant. Colonoscopy after complete discontinuation of prophylactic posttransplant immunosuppression revealed no pathologic findings. These observations imply that host immune dysregulation plays a central role in the perpetuation of IBD. It may be influenced by the implementation of a new allogeneic immune system resulting from the transplantation of hematopoietic stem cells. PMID- 12777868 TI - Nonmyeloablative haploidentical stem-cell transplantation using anti-CD2 monoclonal antibody (MEDI-507)-based conditioning for refractory hematologic malignancies. AB - We initiated a clinical trial of nonmyeloablative haploidentical stem-cell transplantation (SCT) using MEDI-507, an immunoglobulin-G1 monoclonal anti-CD2 antibody. The trial was based on a preclinical major histocompatibility complex mismatched bone marrow transplant model in which graft-versus-host disease (GVHD) was prevented and mixed chimerism as a platform for adoptive cellular immunotherapy was reliably induced. Twelve patients (three cohorts of four patients each) received cyclophosphamide, MEDI-507, and haploidentical unmanipulated bone marrow (n=8) or ex vivo T-cell-depleted peripheral blood stem cells (n=4) for chemorefractory hematologic malignancy. A two-dose regimen and schedule modifications of MEDI-507 were undertaken because of graft loss in the first cohort of four patients and GVHD in the second cohort. With ex vivo T-cell depleted peripheral blood SCT, mixed chimerism occurred in all four patients without GVHD. Two patients, however, subsequently lost their grafts. Nonmyeloablative preparative therapy with MEDI-507 and haploidentical SCT have led to the reliable induction of at least transient mixed chimerism as a potential platform for adoptive cellular immunotherapy. PMID- 12777870 TI - Hepatic artery pseudoaneurysm after liver transplantation: treatment with percutaneous thrombin injection. AB - Pseudoaneurysms of the hepatic artery are a rare complication of liver transplantation. Early diagnosis and treatment are essential to avoid life threatening hemorrhage. Conventional treatment consists of surgical resection and vascular reconstruction or transarterial coil embolization. More recently, percutaneous thrombin injection has been successfully used in the treatment of femoral artery pseudoaneurysms. We describe a 70-year-old woman who had a hepatic artery pseudoaneurysm after orthotopic liver transplantation, which was successfully treated by percutaneous thrombin injection. PMID- 12777869 TI - Molecular signatures of urinary cells distinguish acute rejection of renal allografts from urinary tract infection. AB - Acute rejection (AR) and urinary tract infection (UTI) continue to plague renal transplantation. We tested the hypotheses that UTI does not increase granzyme B mRNA levels in urinary cells, and that the levels distinguish AR from UTI. We measured the levels of granzyme B mRNA in 15 urine specimens from renal allograft recipients with UTI, 29 specimens from patients with AR but without UTI, and 14 specimens from patients without AR and without UTI. We also measured transcript levels in urine specimens from 41 nontransplant individuals, 11 with UTI and 30 without UTI. UTI did not increase granzyme B mRNA levels. Granzyme B mRNA levels were lower in renal allograft recipients with UTI compared with those with AR (P<0.0001). We conclude that bacterial UTI is unlikely to confound AR diagnosis made by measurement of granzyme B mRNA levels in urinary cells. PMID- 12777871 TI - Synergistic immunosuppressive effects of rosmarinic acid and rapamycin in vitro and in vivo. AB - BACKGROUND: We previously demonstrated that rosmarinic acid (RosA) inhibits T cell antigen receptor-induced T-cell activation and proliferation. In this study, we evaluated the ability of RosA alone and in conjunction with currently used immunosuppressive drugs to inhibit in vitro splenic T-cell proliferation and prolong skin graft survival in vivo. METHODS: Mouse splenic T-cell proliferation assays were performed in the presence of RosA alone or in combination with cyclosporine, rapamycin (Rapa), or prednisone (Pred). The in vivo synergistic efficacy of RosA and Rapa was evaluated in the mouse skin allograft model. RESULTS: RosA combined with Rapa or prednisone synergistically inhibited splenic T-cell proliferation, whereas the combination of RosA and cyclosporine additively inhibited T-cell proliferation. The combination of RosA and Rapa synergistically prolonged allograft survival. CONCLUSIONS: We conclude that the combination of RosA and Rapa promotes immunosuppressive effects. PMID- 12777872 TI - Diagnosing PTDM. PMID- 12777873 TI - Pneumonitis associated with the use of mycophenolate mofetil. PMID- 12777875 TI - SA: Re Ross et al. PMID- 12777876 TI - RE: Indo-Asian experience of renal transplantation in Yorkshire: results of a 10 year survey. PMID- 12777877 TI - Therapeutic approaches in the management of oral cyclosporine A intoxication. PMID- 12777878 TI - Shouldn't we finally define posttransplant diabetes? PMID- 12777879 TI - Reversal of severe cholestasis caused by chronic graft-versus-host disease with the MARS liver-support device. PMID- 12777880 TI - Administration of anti-interleukin-2Ralpha monoclonal antibody in bronchiolitis obliterans syndrome after lung transplantation. PMID- 12777881 TI - Posttransplant diabetes mellitus. PMID- 12777882 TI - Bioartificial liver: present and future. PMID- 12777884 TI - Topics in magnetic resonance imaging: sports medicine. PMID- 12777883 TI - Acute liver failure and bioartificial liver support: preventing the former to obviate the latter. PMID- 12777885 TI - Interactive and interventional sports medicine imaging. AB - The concepts and clinical potential of interactive magnetic resonance imaging (MRI) in which an examiner manipulates the joint of interest during MRI, and of interventional sports medicine imaging in which radiological guidance is used for targeted therapy of injuries and masses are described. As illustrated by a series of clinical cases, we believe that with further development, interactive MRI will play an important role in the comprehensive evaluation of patients with shoulder pain or dysfunction as well as other joint derangements. Interventional sports medicine takes advantage of the increasingly sophisticated diagnostic value of MRI and the radiologist's ability to use imaging to guide percutaneous therapy. We review our recent experience treating a wide variety of disorders such as cysts, hematomas, and inflammatory disorders, focusing on techniques utilizing ultrasound and MR guidance. PMID- 12777886 TI - Magnetic resonance imaging of focal articular cartilage lesions. AB - Lesions of the articular cartilage now are recognized as a common, often treatable source of joint disability. Magnetic resonance imaging (MRI) of articular cartilage is a sensitive, noninvasive method for the detection of focal articular cartilage lesions. Advancement in imaging technology now allows for high spatial resolution acquisitions that are able to identify most cartilage lesions, and these acquisitions can be incorporated into everyday clinical imaging protocols. Thus, screening for cartilage abnormalities can be accomplished, along with routine evaluation for ligament and meniscal abnormalities. Familiarity with the appearances of normal cartilage and the full spectrum of cartilage lesions will aid in specific diagnoses. Grading and sizing of cartilage lesions and any underlying bony abnormalities on MRI can help the surgeon in treatment planning; however, some significant cartilage lesions can be difficult to identify and grade by MRI. PMID- 12777887 TI - Magnetic resonance imaging of the hip: sports-related injuries. AB - Normal hip disorders do not account for a large proportion of exercise-related injuries, but they can pose a clinical dilemma because symptoms tend to be nonspecific. Conventional radiographs may demonstrate some causes of hip pain, such as stress fractures and degenerative joint disease. Magnetic resonance imaging (MRI) of the hip has proven valuable in the diagnosis of radiographically occult osseous abnormalities and periarticular soft-tissue disorders such as stress fractures, avulsion injuries, musculotendinous abnormalities, and bursitis. Conventional MRI has been less useful in the evaluation of intra articular lesions, including acetabular labral tears, intra-articular loose bodies, and cartilage lesions. Visualization of intra-articular structures and their abnormalities can be improved by injection of diluted gadolinium, which distends the capsule and leaks into labral tears. This article focuses on the use of conventional radiography and MRI in recreational and professional athletes with painful hip joints. Where possible, MRI is compared with other diagnostic modalities such as bone scan and computed tomography. PMID- 12777888 TI - Magnetic resonance imaging of knee trauma: biomechanical approach. AB - Knee trauma often produces predictable groupings of ligamentous and meniscal injuries. Structures that perform related kinematic functions are damaged by the same traumatic mechanisms. When one supporting structure is disrupted, synergistic structures are jeopardized. Locations of meniscal tear, capsuloligamentous sprain, and osseous injury all provide clues about the mechanism of injury. By understanding the most common patterns of knee injury, a biomechanical approach can be used in the interpretation of magnetic resonance images. The identification of abnormality in one structure should lead to a directed search for subtle abnormalities involving anatomically or functionally related structures, thereby improving diagnostic confidence. PMID- 12777889 TI - Magnetic resonance imaging of sports injuries of the ankle. AB - Basic sports-related injuries of the ankle include ligament tear, tendon degeneration and tear, bone bruise, fracture, impingement, osteochondral defect, and plantar fasciitis. This article discusses the magnetic resonance imaging appearance of these injuries. PMID- 12777890 TI - Sports injuries in the young athlete. AB - Children and adolescents increasingly are participating in competitive sports, and there has been a corresponding rise in the incidence of sports-related injuries in this group. Many sports injuries follow the same patterns as are seen in adults, but the developing musculoskeletal system is uniquely susceptible to other types of injury. This review describes anatomic and developmental aspects that result in increased susceptibility to injury and the magnetic resonance imaging characteristics of immature cartilage, marrow, and menisci. Injuries that are usually or exclusively encountered in children and adolescents are emphasized. PMID- 12777891 TI - Magnetic resonance imaging of sports-related muscle injuries. AB - Magnetic resonance imaging (MRI) is increasingly being used to evaluate muscle injuries because of its unparalleled anatomic resolution and high sensitivity in detecting acute and chronic soft-tissue abnormalities. These features allow detection of characteristic injury patterns that lead to accurate diagnoses and grading of severity. The precise assessment of muscle injuries with MRI plays an important role in determining the treatment plan and prognosis of injured athletes. The article begins with a review of the anatomy and MRI characteristics of normal muscle. The general mechanisms of muscle injury and associated MRI findings are discussed, with emphasis on the most common injuries in both the upper and lower extremities. Several mimics of muscle injury are briefly outlined. PMID- 12777892 TI - Trauma: the paradigm for medical care in the 21st century. PMID- 12777893 TI - Battlefield casualties treated at Camp Rhino, Afghanistan: lessons learned. AB - BACKGROUND: Operation Enduring Freedom is an effort to combat terrorism after an attack on the United States. The first large-scale troop movement (> 1,300) was made by the U.S. Marines into the country of Afghanistan by establishing Camp Rhino. METHODS: Data were entered into a personal computer at Camp Rhino, using combat casualty collecting software. RESULTS: Surgical support at Camp Rhino consisted of two surgical teams (12 personnel each), who set up two operating tables in one tent. During the 6-week period, a total of 46 casualties were treated, and all were a result of blast or blunt injury. One casualty required immediate surgery, two required thoracostomy tube, and the remainder received fracture stabilization or wound care before being transported out of Afghanistan. The casualties received 6 major surgical procedures and 11 minor procedures, which included fracture fixations. There was one killed in action and one expectant patient. The major problem faced was long delay in access to initial surgical care, which was more than 5 hours and 2 hours for two of the casualties. CONCLUSION: Smaller, more mobile surgical teams will be needed more frequently in future military operations because of inability to set up current larger surgical facilities, and major problems will include long transport times. Future improvements to the system should emphasize casualty evacuation, en-route care, and joint operations planning between services. PMID- 12777894 TI - Trauma laparotomy in a rural setting before transfer to a regional center: does it save lives? AB - BACKGROUND: In the rural setting, long distances may necessitate that a patient undergo emergency laparotomy before transfer to a regional trauma center for definitive management. The purpose of this study was to review the experience of three regional trauma centers with such treated patients. METHODS: This study was a retrospective chart review of patients who underwent emergency laparotomy for trauma before transfer, identified from the respective databases of participating centers over a 6-year period. RESULTS: Fifty-six patients met the study criteria. Twenty-six (46%) were transferred primarily for management of the abdominal injury, and 14 of these patients (25% overall) underwent damage control procedures. Overall survival was 82%. Logistic regression demonstrated that transfer for treatment of the extra-abdominal injury was the only significant predictor of survival (odds ratio, 34.33; 95% confidence interval, 1.80-655.24). CONCLUSION: Although patients undergoing laparotomy who were subsequently transferred for management of abdominal injury have reasonable outcome, patients transferred primarily for management of extra-abdominal injury seem to have a survival advantage. PMID- 12777895 TI - Does volume matter? The effect of trauma surgeons' caseload on mortality. AB - BACKGROUND: Evidence suggests that trauma centers treating high volumes of severely injured patients produce lower mortality rates than those with low volumes. However, the effect of individual surgeons' trauma caseload on outcomes has not been studied. This study compares outcomes between high-volume (HV) trauma surgeons admitting many patients with high injury severity, and low-volume (LV) surgeons treating fewer critical patients per year. METHODS: All trauma patients admitted to a large Level I trauma center over a 12-year period were assigned to either the HV or LV group, depending on the yearly volume of their admitting surgeon. Surgeons treating > 35 severely injured (Injury Severity Score > 15) patients per year were considered HV. Student's t test and chi2 analysis were used to test comparability of LV and HV patient groups and to compare mortality rates. Mortality rates of HV and LV surgeons' patients were compared in six injury patterns selected to represent moderate to severe injury. TRISS methodology (z score) was also used to assess outcomes in the two groups. The inherent bias of the TRISS method in comparing trauma outcomes was minimized by the homogeneity of the studied patient population. RESULTS: A total of 16,481 patients were admitted to HV surgeons, and 4,214 patients were admitted to LV surgeons. In all subgroups, HV and LV patients were similar regarding age, sex, physiologic status at admission, injury pattern, and injury severity. Mortality rates for HV and LV surgeons were not significantly different between the two groups in any injury pattern. The z score was 1.88 in the HV patient group versus 0.47 in the LV group. CONCLUSION: Within a single institution, mortality rates for patients treated by surgeons admitting many severely injured patients were not significantly different from low-volume surgeons' patients, although there was a trend toward higher mortality in the less active surgeons' patients in some subgroups. PMID- 12777896 TI - Do factors other than trauma volume affect attrition of ATLS-acquired skills? AB - BACKGROUND: We previously demonstrated that trauma patient volume affects attrition rate of Advanced Trauma Life Support (ATLS)-acquired skills. This study assesses the possible roles of age, gender, and practice specialty on attrition of these skills over 8 years. METHODS: Cognitive (assessed by the 40-item Multiple Choice Question Examination [MCQE]) and clinical (assessed by four trauma Objective Structured Clinical Examination [OSCE] stations) skills performance were compared among physicians who completed the ATLS course 0 months, 6 months, 2 years, 4 years, 6 years, and 8 years previously. The physicians were further divided into the following groups: age < 32 years (n = 72) or 32 years or older (n = 72), gender (41 women and 103 men), and specialty (54 surgeons, 90 nonsurgeons, and 22 general surgeons). Multivariate analysis of variance was used for statistical comparison over time and unpaired t tests for between-group comparisons for each time period, with p < 0.05 being considered statistically significant. RESULTS: Regarding age, MCQE decreased from 82.3 +/- 2.8% to 62.7 +/- 3.0% (mean +/- SD) for age < 32 and from 84.1 +/- 3.6% to 62.8 +/- 2.1% for age 32 or older (p = not significant). Overall OSCE score (maximum, 20) decreased similarly for age < 32 (18.0 +/- 0.4 to 13.6 +/- 2.0) and age > 32 or older (18.0 +/- 0.3 to 12.4 +/- 1.3). Decrease in Priorities and Organized Approach scores also showed no differences between the groups. Regarding gender, MCQE decreased similarly in both groups (women, 81.5 +/- 2.2% to 64.4 +/- 2.4%; men, 83.3 +/- 3.2% to 64.1 +/- 4.2%) and so did OSCE, Priorities, and Organized Approach scores. Regarding specialty, surgeons (83.0 +/- 3.1% to 66.1 +/- 4.5%), nonsurgeons (82.9 +/- 3.2% to 63.3 +/- 3.9%), and general surgeons (82.5 +/- 3.5% to 63.8 +/- 5.3%) showed similar decreases in MCQE scores. Overall OSCE scores and Priority and Approach scores decreased similarly in all specialty groups. When trauma volume was controlled, there was still no difference in attrition rate between surgeons and nonsurgeons. CONCLUSION: Trauma patient volume is the most critical determinant of attrition rate of ATLS-acquired skills. Gender, age (at time of taking the course), and practice specialty do not alter this attrition rate. PMID- 12777897 TI - Complications of preinjury warfarin use in the trauma patient. AB - BACKGROUND: The frequency of use of warfarin anticoagulation increases significantly in the elderly population. It remains controversial whether this puts these patients at increased risk for hemorrhagic complications after trauma. METHODS: We prospectively evaluated consecutive trauma patients who were taking warfarin and compared their outcomes to a group of age-matched patients with head injuries but not taking warfarin. RESULTS: One hundred fifty-nine trauma patients on warfarin were evaluated, 94 (59%) with some type of head trauma; 25 of these 94 patients (27%) had documented intracranial trauma. Fifteen patients died (9.4%); they had an international normalized ratio of 3.3 +/- 1.6 versus 3.0 +/- 2.1 for survivors in the warfarin group (p = 0.585). Twelve deaths were in the group of 25 patients with intracranial injuries (48%). Three patients without head injury died (5%) of other causes not related to warfarin or hemorrhage at a mean of 13 days after admission. Ten of 12 patients on warfarin with intracranial injuries who died had documented loss of consciousness (LOC); two patients who died secondary to an isolated intracranial injury had no LOC. Of 70 age-matched patients with head trauma not taking warfarin, 47 (67%) had intracranial injury and 5 of these died (10%) (p < 0.001 for both values compared with study patients). There were no significant differences for patients with intracranial injury comparing those on warfarin and those who were not in terms of age, gender, mechanism of injury, Injury Severity Score, or Glasgow Come Scale score. CONCLUSION: We conclude that the preinjury use of warfarin does not place the trauma patient at increased risk for fatal hemorrhagic complications in the absence of head trauma. Furthermore, the presence of a head trauma alone is not predictive of mortality. However, the presence of intracranial injury is strongly associated with a mortality rate that is significantly higher than patients with head trauma who are not taking warfarin. LOC is also associated with mortality, but the absence of loss of consciousness does not reliably indicate the absence of intracranial injury or risk of death. PMID- 12777898 TI - Both primary and secondary abdominal compartment syndrome can be predicted early and are harbingers of multiple organ failure. AB - BACKGROUND: Primary abdominal compartment syndrome (ACS) is a known complication of damage control. Recently secondary ACS has been reported in patients without abdominal injury who require aggressive resuscitation. The purpose of this study was to compare the epidemiology of primary and secondary ACS and develop early prediction models in a high-risk cohort who were treated in a similar fashion. METHODS: Major torso trauma patients underwent standardized resuscitation and had prospective data collected including occurrence of ACS, demographics, ISS, urinary bladder pressure, gastric tonometry (GAP(CO2) = gastric regional CO(2) minus end tidal CO(2)), laboratory, respiratory, and hemodynamic data. With primary and secondary ACS as endpoints, variables were tested by uni- and multivariate logistic analysis (MLA). RESULTS: From 188 study patients during the 44-month period, 26 (14%) developed ACS-11 (6%) were primary ACS and 15 (8%) secondary ACS. Primary and secondary ACS had similar demographics, shock, and injury severity. Significant univariate differences included: time to decompression from ICU admit (600 +/- 112 vs. 360 +/- 48 min), Emergency Department (ED) crystalloid (4 +/- 1 vs. 7 +/- 1 L), preICU crystalloid (8 +/- 1 vs. 12 +/- 1L), ED blood administration (2 +/- 1 vs. 6 +/- 1 U), GAP(CO2) (24 +/- 3 vs. 36 +/- 3 mmHg), requiring pelvic embolization (9 vs. 47%), and emergency operation (82% vs. 40%). Early predictors identified by MLA of primary ACS included hemoglobin concentration, GAP(CO2), temperature, and base deficit; and for secondary ACS they included crystalloid, urinary output, and GAP(CO2). The areas under the receiver-operator characteristic curves calculated upon ICU admission are primary= 0.977 and secondary= 0.983. Primary and secondary ACS patients had similar poor outcomes compared with nonACS patients including ventilator days (primary= 13 +/- 3 vs. secondary= 14 +/- 3 vs. nonACS = 8 +/- 2), multiple organ failure (55% vs. 53% vs. 12%), and mortality (64% vs. 53% vs. 17%). CONCLUSION: Primary and secondary ACS have similar demographics, injury severity, time to decompression from hospital admit, and bad outcome. 2 degrees ACS is an earlier ICU event preceded by more crystalloid administration. With appropriate monitoring both could be accurately predicted upon ICU admission. PMID- 12777899 TI - Oxygen debt criteria quantify the effectiveness of early partial resuscitation after hypovolemic hemorrhagic shock. AB - BACKGROUND: The effectiveness of partial resuscitation after hypovolemic hemorrhagic shock with deferment of full resuscitation is critical to successful hypotensive resuscitation. METHODS: To quantitatively address this issue, 40 canines were bled under anesthesia to a mean oxygen debt (O(2)D) of 104 +/- 7.6 mL/kg over 60 minutes (mortality, 40%). Animals surviving the shock were then immediately resuscitated with 0%, 8.4%, 15%, 30%, or 120% (full resuscitation) of shed volume as 5% albumin and held for 2 hours postshock, when the remaining portion of full resuscitation volume was given. Animals were followed for 7 days postshock with hepatic and renal function studies, and then, under anesthesia, cardiac output and organ biopsy specimens were taken before the animals were killed. RESULTS: By 2 hours postshock, 0% immediate resuscitation had an O(2)D increase of 80 mL/kg above end of shock, but O(2)D at 8.4% immediate resuscitation decreased -30 mL/kg, 15% immediate resuscitation fell -65 mL/kg, 30% immediate resuscitation decreased -80 mL/kg below end of shock, and O(2)D with 120% full resuscitation fell to preshock levels. All decreases in O(2)D were significantly (p < 0.05) below end of shock, but both 15% and 30% immediate resuscitation exceeded the 8.4% immediate resuscitation rate (p < 0.05) throughout the resuscitation, and 120% full resuscitation exceeded these (p < 0.05). The immediate resuscitation O(2)D response correlated significantly (p < 0.001) with base deficit and lactate, but blood pressure was not a significant discriminator. Seven-day biopsies showed return of bowel mucosa but a pattern of cellular injury in heart, liver, and kidney that improved from 8.4% < 15% < 30 < 120% immediate resuscitation. CONCLUSION: The data suggest that, compared with 120% postshock immediate resuscitation, 8.4% and 15% immediate resuscitation give poorer results, with 30% immediate resuscitation showing mild, transient, but acceptable changes in organ function allowing for a 2-hour delay until full resuscitation, with complete 7-day recovery. Base deficit and lactate, but not blood pressure, are significant indices of O(2)D. PMID- 12777900 TI - LF 16-0687 Ms, a new bradykinin B2 receptor antagonist, improves neurologic outcome but not brain tissue prostaglandin E2 release in a rat model of closed head trauma combined with ethanol intoxication. AB - BACKGROUND: LF 16-0687 Ms previously was reported to improve Neurological Severity Score (NSS) and decrease cerebral edema and prostaglandin E(2) (PGE(2)) release after closed head trauma (CHT) in rats. Here, we examined whether these beneficial effects of LF 16-0687 Ms are altered when CHT is accompanied by acute ethanol administration. METHODS: Six groups of rats (n = 8 per group) were examined during combination of the following experimental conditions: CHT versus sham operation, LF 16-0687 Ms 3 mg/kg subcutaneously versus saline, and ethanol 2 g/kg versus saline. RESULTS: After CHT, brain water content decreased and NSS improved with ethanol + LF 16-0687 Ms as compared with values after saline or ethanol. PGE(2) release decreased with ethanol (147 +/- 59 pg/mg tissue) but not with ethanol + LF 16-0687 Ms (286 +/- 194 pg/mg tissue). CONCLUSION: Ethanol does not affect the improvement of NSS and the decrease of cerebral edema seen with LF 16-0687 Ms after CHT, but does reverse the ability of LF 16-0687 Ms to minimize the increase of PGE(2) release. In intoxicated patients, bradykinin antagonist therapy may improve post-CHT outcome without altering PGE(2) release. PMID- 12777901 TI - Can within-category naming identify subtle cognitive deficits in the mild traumatic brain-injured patient? AB - BACKGROUND: This investigation examined the effect of a speeded, computer controlled task on detecting differences in latency and accuracy of within category name generation in adults having sustained a mild traumatic brain injury (MTBI). METHODS: Twenty-four adults in acute recovery and 24 age-matched controls were instructed to view 72 pictures on a computer monitor, and then name another item belonging to the same category as the visual stimulus as quickly as possible. RESULTS: The MTBI group demonstrated significantly longer latencies (p < 0.001) and lower accuracy (p < 0.001) than the control group. Both groups displayed similar patterns of response, although the MTBI group produced significantly more perseverative errors (p < 0.001). No significant correlations were found between performance on the Scales of Cognitive Ability for Traumatic Brain Injury and response latency or accuracy. CONCLUSION: The MTBI group performed the task significantly slower and less accurately than controls. Reaction time measures may prove more sensitive than traditional assessment measures in detecting subtle difficulties. PMID- 12777903 TI - Allogeneic blood transfusion increases the risk of postoperative bacterial infection: a meta-analysis. AB - BACKGROUND: Immunosuppression is a consequence of allogeneic (homologous) blood transfusion (ABT) in humans and is associated with an increased risk in cancer recurrence rates after potentially curative surgery as well as an increase in the frequency of postoperative bacterial infections. Although a meta-analysis has been reported demonstrating the relationship between ABT and colon cancer recurrence, no meta-analysis has been reported demonstrating the relationship of ABT to postoperative bacterial infection. METHODS: Twenty peer-reviewed articles published from 1986 to 2000 were included in a meta-analysis. Criteria for inclusion included a clearly defined control group (nontransfused) compared with a treated (transfused) group and statistical analysis of accumulated data that included stepwise multivariate logistic regression analysis. In addition, a subgroup of publications that included only the traumatically injured patient was included in a separate meta-analysis. A fixed effects analysis was conducted with odds ratios obtained by using the conditional maximum likelihood method and 95% confidence intervals on the obtained odds ratios were determined using the mid-p technique. RESULTS: The total number of subjects included in this meta-analysis was 13,152 (5,215 in the transfused group and 7,937 in the nontransfused group). The common odds ratio for all articles included in this meta-analysis evaluating the association of ABT to the incidence of postoperative bacterial infection was 3.45 (range, 1.43-15.15), with 17 of the 20 studies demonstrating a value of p < or = 0.05. These results provide overwhelming evidence that ABT is associated with a significantly increased risk of postoperative bacterial infection in the surgical patient. The common odds ratio of the subgroup of trauma patients was 5.263 (range, 5.03-5.43), with all studies showing a value of p < 0.05 (0.005 0.0001). These results demonstrate that ABT is associated with a greater risk of postoperative bacterial infection in the trauma patient when compared with those patients receiving ABT during or after elective surgery. CONCLUSION: These results demonstrate that ABT is an associated and apparently significant and frequently overlooked risk factor for the development of postoperative bacterial infection in the surgical patient. Allogeneic blood transfusion is a greater risk factor in the traumatically injured patient when compared with the elective surgical patient for the development of postoperative bacterial infection. PMID- 12777902 TI - Blood transfusion, independent of shock severity, is associated with worse outcome in trauma. AB - BACKGROUND: We have previously shown that blood transfusion in the first 24 hours is an independent predictor of mortality, intensive care unit (ICU) admission, and increased ICU length of stay in the acute trauma setting when controlling for Injury Severity Score, Glasgow Coma Scale score, and age. Indices of shock such as base deficit, serum lactate level, and admission hemodynamic status (systolic blood pressure, heart rate) and admission hematocrit were considered potential confounding variables in that study. The objectives of this study were to evaluate admission anemia and blood transfusion within the first 24 hours as independent predictors of mortality, ICU admission, ICU length of stay (LOS), and hospital LOS, with serum lactate level, base deficit, and shock index (heart rate/systolic blood pressure) as covariates. METHODS: Prospective data were collected on 15,534 patients admitted to a Level I trauma center over a 3-year period (1998-2000) and stratified by age, gender, race, Glasgow Coma Scale score, and Injury Severity Score. Admission anemia and blood transfusion were assessed as independent predictors of mortality, ICU admission, ICU LOS, and hospital LOS by logistic regression analysis, with base deficit, serum lactate, and shock index as covariates. RESULTS: Blood transfusion was a strong independent predictor of mortality (odds ratio [OR], 2.83; 95% confidence interval [CI], 1.82 4.40; p < 0.001), ICU admission (OR, 3.27; 95% CI, 2.69-3.99; p < 0.001), ICU LOS (p < 0.001), and hospital LOS (Coef, 4.37; 95% CI, 2.79-5.94; p < 0.001) when stratified by indices of shock (base deficit, serum lactate, shock index, and anemia). Patients who underwent blood transfusion were almost three times more likely to die and greater than three times more likely to be admitted to the ICU. Admission anemia (hematocrit < 36%) was an independent predictor of ICU admission (p = 0.008), ICU LOS (p = 0.012), and hospital LOS (p < 0.001). CONCLUSION: Blood transfusion is confirmed as an independent predictor of mortality, ICU admission, ICU LOS, and hospital LOS in trauma after controlling for severity of shock by admission base deficit, lactate, shock index, and anemia. The use of other hemoglobin-based oxygen-carrying resuscitation fluids (such as human or bovine hemoglobin substitutes) in the acute postinjury period warrants further investigation. PMID- 12777904 TI - Resuscitation with a novel hemoglobin-based oxygen carrier in a Swine model of uncontrolled perioperative hemorrhage. AB - BACKGROUND: Systemic and pulmonary hypertension, possibly related to nitric oxide scavenging by free hemoglobin (Hb), is often seen during resuscitation with hemoglobin-based oxygen carriers (HBOCs). Recently, a second-generation HBOC, rHb2.0 for Injection (rHb), has been developed using recombinant human Hb that has reduced reactivity with nitric oxide. The current study evaluates the efficacy of this novel compound for resuscitation in a swine model of uncontrolled perioperative hemorrhage. METHODS: After instrumentation, animals underwent splenectomy and rapid hemorrhage to a systolic blood pressure of 35 mm Hg and isoelectric electroencephalography. 15 minutes of shock was followed by resuscitation over 30 minutes. In phase I, 18 animals were randomized into three resuscitation groups: (1) lactated Ringer's (LR) equal to three times the shed blood, the negative control group; (2) heterologous blood (BL) equal to Hb 2 g/kg, the positive control group; and (3) rHb equal to 2 g/kg, the treatment group. In phase II, six animals underwent the same experiment with a first generation HBOC, diaspirin cross-linked Hb (DCLHb) equal to 2 g/kg, an additional control group. On day 0 after 2 hours of observation, spontaneously breathing animals were returned to their cages. Surviving animals were redosed on days 1, 2, and 3 (rHb/DCLHb 1 g/kg; LR/BL-LR 500 mL). Survivors were killed on day 5 and organs harvested for histologic examination. Group comparisons were performed using Student's t test, repeated-measures analysis of variance, and chi2 test. Significance was set at 95% confidence intervals. RESULTS: After resuscitation, systemic mean arterial pressure (MAP) (baseline = 107 +/- 15 mm Hg) was 128 +/- 34 and 108 +/- 15 mm Hg in rHb and BL animals, respectively, and remained stable. In LR and DCLHb animals, after normalization, MAP declined to 67 +/- 13 and 84 +/ 34 mm Hg, respectively. The rHb group maintained higher MAP than the LR and BL groups (p < 0.05 vs. both). With resuscitation, mean pulmonary arterial pressure (PAP) (baseline = 25 +/- 5 mm Hg) increased in rHb (40 +/- 4 mm Hg), BL (34 +/- 3 mm Hg), and DCLHb (40 +/- 3 mm Hg) groups, but stayed elevated only in the DCLHb group (36 +/- 3 mm Hg). PAP in the rHb group was similar to the BL group (p > 0.05), and both rHb and BL groups showed a higher PAP than the LR group (p < 0.05 vs. both). PAP was highest in the DCLHb group (p < 0.05 vs. rHb). Cardiac output of rHb and BL groups was similar (p > 0.05) throughout the observation period. Arterial lactate increased to 5.6 +/- 2.5 mmol/L with shock and then normalized to < 2.0 mmol/L in the rHb, BL, and LR groups within 30 minutes of resuscitation. It remained elevated to > 3.5 mmol/L and showed a delayed increase in the DCLHb group (p < 0.05). Causes and number of deaths were as follows: rHb, zero of six; BL-transfusion reaction, one of six; LR-irreversible shock, four of six; and DCLHb-ventricular failure, six of six. There was no significant increase in plasma methemoglobin (rHb) and no difference in liver or cardiac enzymes (rHb vs. BL). No histologic abnormalities were seen in the rHb group except for cytoplasmic vacuolation, a process thought to be related to metabolism of the test article. CONCLUSION: rHb2.0 for Injection, a second-generation recombinant human HBOC, performs as well as heterologous blood for resuscitation after perioperative blood loss, does not cause sustained pulmonary hypertension, maintains adequate cardiac output and oxygen delivery, and is superior to either LR or DCLHb. PMID- 12777905 TI - Multiplicity of solid organ injury: influence on management and outcomes after blunt abdominal trauma. AB - OBJECTIVE: The current study was undertaken to examine how concomitant injury to liver and spleen after blunt abdominal trauma affects management and outcomes. METHODS: This study was a retrospective chart review of all blunt abdominal trauma patients admitted with a diagnosis of liver or spleen injury at two Level I trauma centers over a 4-year period. Presentation, injury grade, management, and outcomes were analyzed. Patients with single-organ injury (liver or spleen) were compared with patients having injury to both organs (liver and spleen). Significance was set at 95% confidence intervals. RESULTS: Of 1,288 patients who met entry criteria, 1,125 had single (spleen, 573; liver, 552) organ injury (group S) and 163 had injury to both organs (group B). Group B patients had significantly higher Injury Severity Score, higher admission lactate, and lower admission systolic blood pressure and base excess. Eighty-one percent (915 of 1,125) of group S and 69% (112 of 163) of group B patients were managed nonoperatively (p < 0.05). Of the nonoperatively managed patients, 5.8% (53 of 915) in group S and 11.6% (13 of 112) in group B failed this form of therapy (p < 0.05). Higher failure rate in group B was because of bleeding from injured solid organ(s), and not non-solid organ related failures. Mortality, intensive care unit and hospital lengths of stay, and transfusion requirements were all significantly higher in group B. CONCLUSION: Blunt trauma patients with concomitant injury to liver and spleen have higher Injury Severity Score, mortality, lengths of stay, and transfusion requirements. There is a higher failure rate with nonoperative management, and therefore extra vigilance is warranted when choosing this form of therapy in the presence of injury to both organs. PMID- 12777906 TI - The effect of implant overlap on the mechanical properties of the femur. AB - BACKGROUND: The most biomechanically stable relationship between the side plate of a compression hip screw (CHS) and retrograde intramedullary (IM) femoral nail has not been described in the literature. This becomes a clinical issue when treating supracondylar femur fractures with a retrograde nail in patients with a history of compression hip screw fixation of intertrochanteric fractures. The proximal end of the nail and the interlocking screws may act as a stress riser in the femoral diaphysis. The purpose of this study is to determine the biomechanical consequences of the amount of implant overlap between a CHS plate and retrograde IM femoral nail. METHODS: Nine paired fresh-frozen cadaver femora from elderly donors were cleaned of soft tissue and fixed with uniaxial strain gauges. Each femur was loaded three times in a fall-loading configuration to 50 kg at a rate of 1 Hz. The study consisted of two phases. In phase 1, six pair were randomly divided into a control and test femur from each pair. Three states were compared on each test femur: uninstrumented, instrumented with CHS, and instrumented with CHS and retrograde nail. The control femur consisted of the matched femur tested in two states: uninstrumented and instrumented with a CHS. The femora were then loaded to failure. The tests were performed with the retrograde nail and CHS gapped 3 cm, kissing, and overlapping by two screw holes (two pair for each state). In phase 2, each of the remaining three pair were instrumented with a CHS and retrograde nail overlapping in one femur and gapped in the matched femur and tested in the same manner. Data analysis was performed using Pearson's correlation coefficients between groups. Paired samples t tests were used to compare differences within test states and independent samples t tests were used to compare differences between femora. Mean strain at 50-kg load, load-versus-strain patterns, failure patterns, and load and strain at failure were recorded. RESULTS; Correlation coefficients were greater than 0.98 within and between pairs (p < 0.001). There were statistically significant differences (p < 0.05) in strain patterns between the uninstrumented, CHS, and CHS/IM test states. The addition of a side plate significantly (p < 0.05) increased lateral compressive strains in the femoral diaphysis. Mean strain at 50-kg load was significantly (p < 0.05) altered by the addition of the retrograde nail in all three implant orientations. Gapped implants failed at lower loads and strains than femurs with kissing and overlapping implants. Gapped constructs failed at lower loads than control states. Overlapped constructs tolerated the highest loads and strains before failure. CONCLUSION: Strain patterns are altered by the degree of implant overlap in the proximal femoral diaphysis. Femora with uninstrumented intervals between retrograde nails and side plates fail at lower loads than femora without retrograde nails and those with kissing or overlapping implants. Kissing or overlapping instrumentation increases load to failure and creates a more biomechanically stable construct than gapped implants. The findings of this study suggest an overlapping implant orientation in the femur increases failure load at the implant interface. PMID- 12777907 TI - Extracorporeal shock waves in the treatment of nonunions. AB - BACKGROUND: Nonunion remains a major complication after skeletal trauma. In the last decade, extracorporeal shock wave therapy has become a common tool for the treatment of nonunions. To date, no prospective, randomized trial has been conducted to show the efficacy of this form of treatment. METHODS: This study was performed to determine the value of extracorporeal shock wave therapy for nonunions. Previous published results in the literature and our own clinical results were analyzed and related to the natural history of bony union. RESULTS: No study has proven that extracorporeal shock wave therapy improves bone healing. Clinical studies reporting the acceleration of union after application of shock waves instead seem to misinterpret the natural history of bony union. CONCLUSION: No evidence supports the treatment of pseudarthroses with extracorporeal shock waves. A randomized, prospective, clinical trial with a control group has to be performed before a final decision can be made regarding this indication for extracorporeal shock wave therapy. PMID- 12777908 TI - Complications of locked nailing in humeral shaft fractures. AB - BACKGROUND: The purpose of this study was to investigate the complications of humeral locked nailing. METHODS: Between 1994 and 2000, 161 humeral shaft fractures (98 acute fractures and 63 delayed unions or nonunions) in 159 patients treated with humeral locked nails were followed up for an average of 25.4 months. There were 89 men and 70 women, with an average age of 53.5 years. Acute fractures included 68 closed, 18 type I, 8 type II, 3 type IIIA, and 1 type IIIB open fractures. Thirty-six nonunions had previous operations. In general, acute fractures were treated with closed nailing and nonunions were treated with open nailing with bone grafting. Since 1998, interfragmentary wiring has been added in nonunions to compress the fracture. RESULTS: In total, 30 patients had 31 significant complications. Nine of them were persistent nonunions, six from acute fractures and three from nonunions. Fracture gap was associated with a significantly higher risk of nonunion. The risk of operative comminution was significantly higher in retrograde nailing, and operative comminution resulted in a significantly higher risk of nonunion. Seven of the nine nonunions underwent revisional nailing and achieved eventual union. Removal of the protruded screws was performed in two cases. Other complications included shoulder impairment, elbow impairment, angular malunion, and post-nailing radial nerve palsy. CONCLUSION: Many complications of humeral locked nailing can be prevented by improving the implant design or surgical techniques. The patients with persistent nonunion can be reliably treated by revisional nailing and bone grafting. PMID- 12777909 TI - Clinical effects of inhibiting leukocyte adhesion with monoclonal antibody to intercellular adhesion molecule-1 (enlimomab) in the treatment of partial thickness burn injury. AB - BACKGROUND: A randomized, prospective, multicenter, double-blind, placebo controlled, phase II clinical trial was performed to determine whether inhibition of leukocyte adherence by administration of monoclonal antibody directed against intercellular adhesion molecule-1 would improve burn wound healing. METHODS: One hundred ten patients with burn injury ranging from 10% to 30% total body surface area were enrolled. Fifty-six patients received placebo (saline) and 54 patients received murine monoclonal antibody to the human intercellular adhesion molecule 1 (enlimomab). Treatment was initiated within 6 hours of injury. Patients had three distinct partial-thickness wound sites assessed. Laser Doppler flowmetry was used to stratify wounds on the day of injury. Wounds were assessed for healing status on day 21 postburn and categorized as healed, nonhealed, or grafted. RESULTS: Patients treated with enlimomab had a significantly increased percentage of wounds that healed spontaneously in less than 21 days overall and when stratified by burn wound laser Doppler blood flow readings for those wounds at greatest risk for nonhealing. CONCLUSION: These results support the concept that leukocyte adherence is involved in the pathogenesis of burn wound necrosis and suggest a therapeutic mechanism for modulating the inflammatory response after the burn injury that may improve wound healing. PMID- 12777910 TI - The risk factors and time course of sepsis and organ dysfunction after burn trauma. AB - BACKGROUND: Sepsis and organ dysfunction are common and likely contribute to death after burn trauma. We sought to define relationships between sepsis, severe multiple organ dysfunction (MOD), and death after burn trauma. METHODS: Adults with > or = 20% total body surface area burns were prospectively enrolled. Information regarding infection, severity of sepsis, and organ failure was collected daily. Risk factors (e.g., age, burn size, shock) were analyzed for their association with severe MOD, complicated sepsis, and death. We characterized the temporal relationship between organ failure and sepsis. RESULTS: Of 175 patients, 27% developed severe MOD, 17% developed complicated sepsis, and 22% died. Full-thickness burn size, age, and inhalation injury were associated with MOD, sepsis, and death. Infection preceded MOD in 83% of patients with both. A base deficit of > or = 6 mEq/L at 24 hours after injury was associated with death. CONCLUSION: When it occurs, severe MOD is usually preceded by infection. In addition, an elevated base deficit at 24 hours and septic shock are the most important factors associated with and possibly contributing to death after burn trauma. PMID- 12777911 TI - The risk of death for Jehovah's Witnesses after major trauma. AB - BACKGROUND: Trauma surgeons are faced with life-threatening blood loss in patients such as Jehovah's Witnesses. We assessed and compared the risks of death after major trauma for Jehovah's Witnesses and other religious groups. METHODS: A retrospective cohort study was conducted between August 1992 and September 1999 in a Level I academic trauma center. Statistical methods included Tukey's one-way analysis of variance, chi2 analysis, and bivariate and multivariate logistic regression analyses. RESULTS: The cohort consisted of 556 patients: 82 Jehovah's Witnesses (14.7%), 52 Baptists (9.4%), 101 Catholics (18.2%), and 321 patients belonging to other religious groups (57.7%). Mean Injury Severity Scores for 433 patients were 10.3 +/- 9, 8.9 +/- 10, 10.3 +/- 11, and 11.3 +/- 14, respectively. There were no significant differences in mean Injury Severity Scores between religious groups, and no statistically significant associations between religion and Injury Severity Scores were identified. Significant predictors of mortality were age, systolic blood pressure at admission, Glasgow Coma Scale score, and type of trauma. Jehovah's Witnesses were 6% more likely to die after major trauma than Baptists, 20% more likely than Catholics, and as likely as patients from any other religious groups. CONCLUSION: After controlling for age, race, systolic blood pressure, Glasgow Coma Scale score, and type of trauma, Jehovah's Witnesses have a nonsignificant increased risk of death after major trauma compared with other religious groups. PMID- 12777912 TI - Quality of multiple trauma care in 33 German and Swiss trauma centers during a 5 year period: regular versus on-call service. AB - BACKGROUND: The purpose of this study was to evaluate process and outcome quality of severely injured patients admitted during on-call (OC) versus regular trauma service (RS). METHODS: This was a prospective and multicentric analysis of the Trauma Registry of the German Trauma Society. Patients were evaluated if directly admitted from the scene of accident with an Injury Severity Score of > 15 and if alive on arrival at the emergency department. RESULTS: Seventy percent of patients were admitted during OC; these patients were significantly younger. Blunt trauma predominated, with a 95% incidence. Falls from great heights were significantly more frequent during RS, whereas motor vehicle crashes predominated during OC. No differences were found for emergency department management (e.g., time to abdominal ultrasound, chest radiograph, or cranial computed tomography). However, time to admission to the intensive care unit was substantially longer during RS. No significant differences were found for outcome parameters such as length of intensive care unit stay, hospitalization time, incidence of organ failure, or mortality. CONCLUSION: This study demonstrates a constant quality of care provided 24 hours per day, 7 days per week in the participating hospitals. Differences within individual trauma centers were not compared and need to be assessed by internal quality management. PMID- 12777913 TI - Immediate microcirculatory derangements in skeletal muscle and periosteum after closed tibial fracture. AB - BACKGROUND: Severe musculoskeletal soft tissue injury sustained after a closed fracture to the extremities significantly influences bone healing and determines the patient's prognosis. The present study was aimed at quantitatively assessing immediate microcirculatory changes in skeletal muscle and periosteum after standardized closed fracture. METHODS: Standardized closed fracture of the left tibia in isoflurane-anesthetized Sprague-Dawley rats (n = 14) was induced using a modified weight-drop technique. The left extensor digitorum longus (EDL) muscle (n = 7) and tibial periosteum (n = 7) were surgically exposed for in vivo fluorescence microscopy 15 minutes after fracture. Nonfractured rats (n = 14) served as controls. EDL muscle edema was determined by the ratio of wet to dry weight (EDL water content). RESULTS: Closed tibial fracture resulted in a significant reduction of functional capillary density, red blood cell velocity, and volumetric blood flow in both EDL muscle and periosteum. Microvascular diameter, leukocyte adherence, and macromolecular leakage were markedly increased, indicating trauma-induced inflammation and endothelial disintegration. EDL muscle edema was found increased significantly after fracture. CONCLUSION: This model permits for the first time direct in vivo visualization and quantification of fracture-induced microhemodynamic changes and cellular interactions within the surrounding soft tissue. It demonstrates that even simple fractures lead to profound microcirculatory disturbances in skeletal muscle and periosteum, and also at sites remote from the diaphyseal fracture site. It provides a useful approach for the development of therapeutic strategies to counteract fracture-induced microvascular dysfunction. PMID- 12777914 TI - Single dose of acetylsalicylic acid prevents thromboxane release after tourniquet ischemia. AB - BACKGROUND: Ischemia, such as that caused by a tourniquet, stimulates thromboxane (Tx) A(2) synthesis. TxA(2) might sensitize the operated limb to various complications, such as compartment syndrome and thromboembolic events. METHODS: We studied the effect of pretreatment with a single dose of acetylsalicylic acid (ASA) (25, 100, and 500 mg) given 3 hours before surgery on the formation of TxB(2), a stable metabolite of TxA(2), after tourniquet deflation in 32 knee or ankle surgery patients. RESULTS: Tourniquet time varied between 60 +/- 8 to 71 +/ 7 (SE) minutes. In control patients without ASA pretreatment, the platelet produced femoral vein serum TxB(2) concentration over 30 minutes in vitro coagulation increased remarkably (from 40.0 +/- 20 ng/mL to 73.5 +/- 39 ng/mL) immediately after tourniquet deflation. Plasma concentrations increased similarly, approximately threefold. Pretreatment with 100 or 500 mg ASA prevented the increase in TxB(2) concentrations. Radial artery concentrations of TxB(2) were similar to venous concentrations in the different treatment groups. CONCLUSION: Pretreatment with a single 100-mg dose of ASA prevents the release of TxB(2) after tourniquet deflation. PMID- 12777915 TI - Readiness to change alcohol use after trauma. AB - BACKGROUND: Alcohol is the leading risk factor for severe injury. This study examined whether patients hospitalized after an alcohol-related injury are motivated to change alcohol use, thus making them potential candidates for brief motivational interventions. METHODS: Fifty patients hospitalized in a Level I trauma center, admitted with a positive blood alcohol concentration, were assessed for motivation to change alcohol-related behavior using validated questionnaires. Information was gathered regarding level of alcohol use, consequences of use, and motivation to change drinking habits. Demographic variables, alcohol use measures, perception of alcohol's contribution to the current injury, and negative consequences of use were evaluated by linear regression to predict readiness to change drinking. RESULTS: Mean blood alcohol concentration was 197 mg/dL at admission. Patients reported a pattern of binge drinking, with 86% reporting at least one binge-drinking episode in the past month, and a mean of 3.4 days of binge drinking per month. Most patients (84%) reported considering making a change (cutting down or quitting) in their drinking. Finally, patients reported experiencing an average of 22.5 negative lifetime consequences to their drinking. Having more negative consequences was found to significantly predict readiness to change drinking (p < 0.001). CONCLUSION: In this study, most patients were motivated to change their drinking. An increased number of negative consequences of alcohol use before admission predicted readiness to change drinking habits. Brief motivational interventions would be a reasonable option in this group of patients. PMID- 12777916 TI - Impact of culture and policy on organ donation: a comparison between two urban trauma centers in developed nations. AB - BACKGROUND: The similarities and differences in organ donation policies, consent rates, and number of organs transplanted from patients declared "brain dead" after traumatic injury in different countries has not been previously reported. METHODS: An international trauma survey questionnaire was developed. Analysis of two responding centers with regard to organ donation practices between urban, free-standing adult trauma hospitals is presented: the R Adams Cowley Shock Trauma Center (STC) in Baltimore, Maryland, and the Lorenz Bohler Hospital (LBH) in Vienna, Austria. RESULTS: Hospital admissions resulting from traumatic brain injury (TBI) account for a significantly greater number of admissions at the STC than at the LBH (761 vs. 276), and the STC has a higher number of patients admitted with severe TBI (Glasgow Coma Scale score < 8). Of 39 medically suitable brain-dead patients at the STC, 18 went to organ donation. At the LBH, 16 patients were declared brain-dead, 7 were considered to be medically suitable, and all 7 went to donation. A "presumed consent" organ donation policy in Austria resulted in 100% of medically suitable patients going to donation at the LBH. With a volunteer donation policy at the STC, 46% of patients went to donation. Of those families who refused donation at the STC, 9 of 16 eligible African Americans (56%), 10 of 21 eligible Caucasians (48%), 1 Hispanic, and 1 Native American Indian family declined donation. CONCLUSION: "Presumed" organ donation in Austria led to 4 organs transplanted per trauma brain-death at the LBH, as compared with 3.8 organs per brain-death at the STC. The greater number of patients with severe TBI at the STC accounts for a similar organ donation rate compared with the LBH, despite the fact that the consent at the STC is voluntary and at the LBH is "presumed." A higher organ donation rate in the United States would result in a greater number of organ transplants from patients who die after traumatic injury and a resultant increase in potential lives saved. There does not appear to be a significant difference in ethnicity between families who accept and those who refuse organ donation after traumatic brain death declaration at the STC. PMID- 12777917 TI - Percutaneous transluminal coronary angioplasty in a patient with myocardial infarction after penetrating trauma. PMID- 12777918 TI - Traumatic multiple pseudoaneurysms of the intrasplenic artery: case report. PMID- 12777919 TI - Traumatic bilateral testicular dislocation. PMID- 12777920 TI - Management of severe combined pulmonary and myocardial contusion with extracorporeal membrane oxygenation. PMID- 12777921 TI - Isolated pulmonary mucormycosis in a patient with multiple injuries: a case presentation and review of the literature. PMID- 12777922 TI - Incorporating sustainability into the concept of optimal care. PMID- 12777923 TI - Craniofacial and skull base trauma. AB - BACKGROUND: Traumatic craniofacial and skull base injuries require a multidisciplinary team approach. Trauma physicians must evaluate carefully, triage properly, and maintain a high index of suspicion to improve survival and enhance functional recovery. Frequently, craniofacial and skull base injuries are overlooked while treating more life-threatening injuries. Unnoticed complex craniofacial and skull base fractures, cerebrospinal fluid fistulae, and cranial nerve injuries can result in blindness, diplopia, deafness, facial paralysis, or meningitis. Early recognition of specific craniofacial and skull base injury patterns can lead to identification of associated injuries and allow for more rapid and appropriate management. CONCLUSION: Early detection and treatment of craniofacial and skull base traumatic injuries should lead to decreased morbidity and mortality. This review discusses the most common of these injuries, their possible complications, and treatment. PMID- 12777924 TI - Impaled fractured rib. PMID- 12777925 TI - Thoracic plank impalement: an engineering perspective. PMID- 12777926 TI - Reducing future combat deaths through medical technology. PMID- 12777927 TI - Empty sella and exposure to radio-frequency fields. PMID- 12777928 TI - Health care economic analyses. AB - Health care economic analyses are becoming increasingly important as health care costs consume an increasing proportion of the gross domestic product (GDP). Among the different forms of health care economic analysis are (1) cost-minimization analysis, (2) cost-benefit analysis, (3) cost-effectiveness analysis, and (4) cost-utility analysis. Cost-utility analysis is the most sophisticated because it incorporates the value (improvement in quality of life and length of life) conferred by an intervention for the resources expended. The different types of analyses of importance to the clinician are addressed herein. PMID- 12777929 TI - Advances in the development of visual prostheses. AB - Visual prostheses are based on neuronal electrical stimulation at different locations along the visual pathway (ie, cortical, optic nerve, epiretinal, subretinal). In terms of retinal prostheses, advances in microtechnology have allowed for the development of sophisticated, high-density integrated circuit devices that may be implanted either in the subretinal or epiretinal space. Analogous to the cochlear implants for some forms of deafness, these devices could restore useful vision by converting visual information into patterns of electrical stimulation that would excite the remaining spared inner retinal neurons in patients with diseases such as retinitis pigmentosa and age-related macular degeneration. The different types of implants and recent results are discussed, but special emphasis is given to retinal implants. PMID- 12777930 TI - New treatments for serpiginous choroiditis. AB - Serpiginous choroiditis is a bilateral, inflammatory condition of the inner choroid and overlying retinal pigment epithelium with a distinctive retinal distribution. Although rare, serpiginous choroiditis deserves attention because it can lead to permanent loss of vision due to foveal involvement by inflammation or choroidal neovascularization from frequent recurrences. This article reviews the recent ophthalmic literature published on the management of serpiginous choroiditis. PMID- 12777931 TI - Cost-effectiveness of ocular interventions. AB - The cost-effectiveness of ophthalmic interventions has become increasingly important as the US health care budget continues to grow. During the past two decades, a methodology rooted in utility analysis has provided a way to objectively assess the value of an intervention for a specific disease. Specifically within the field of ophthalmology, there is a growing body of literature on utility analysis using patient preference-based outcomes for ophthalmic disease. This has led the way to combining costs with utility values to derive a cost-utility analysis, or cost-effectiveness, for a given ophthalmic procedure. This article outlines this methodology, provides a reference-case example for initial cataract surgery, and describes a comparative study of the value associated with interventions within the field of ophthalmology and across other medical specialties. PMID- 12777932 TI - Update on treatment of retinal arterial occlusions. AB - Retinal arterial occlusion remains a difficult clinical entity to manage. Treatment can best be categorized as conservative (ocular massage, pharmacologic, anterior chamber paracentesis) and invasive (catheterization of the proximal ophthalmic artery through the femoral artery with the infusion of thrombolytic agents). Most reports remain anecdotal due to its low incidence (0.85/100,000/y). PMID- 12777933 TI - Circumscribed choroidal hemangioma. AB - Circumscribed choroidal hemangioma is an uncommon, benign vascular tumor manifesting as an orange-red mass in the posterior pole of the eye. Serous retinal detachment accounts for decreased vision in most patients. Diagnosis of this tumor is challenging with many patients initially misdiagnosed with choroidal melanoma or metastasis. Several ancillary tests such as ultrasonography, fluorescein angiography, indocyanine green angiography, and magnetic resonance imaging help differentiate this tumor from other simulating lesions. Asymptomatic lesions should be observed, but visually threatening or visually impairing lesions require treatment. Photodynamic therapy, laser photocoagulation, and transpupillary thermotherapy may be used for primary management of this tumor. Patients who fail to respond to previous treatment or those with extensive serous retinal detachment can be treated using radiotherapeutic modalities. Long interval between onset of symptoms and treatment, poor visual acuity at presentation, and presence of chronic retinal or retinal pigment epithelial changes are associated with poor long-term vision. PMID- 12777934 TI - Cataract surgery results after pars plana vitrectomy. AB - Continuous refinements in vitreoretinal surgical techniques and an increasing number of posterior segment disorders are being successfully managed with pars plana vitrectomy (PPV). This has resulted in an increase in the number of vitrectomized eyes seen by cataract surgeons. Cataract surgery in previously vitrectomized eyes has been reported to be more challenging than in eyes without previous vitrectomy. Special considerations and precautions are often required before, during, and after the surgery to successfully address these challenges. Several studies have reported that phacoemulsification with intraocular lens implantation after PPV is a relatively safe procedure that can improve visual acuity and quality of life. The extent of visual improvement may be limited only by retinal comorbidity. PMID- 12777935 TI - Current indications of transpupillary thermotherapy for the treatment of posterior segment diseases. AB - Transpupillary thermotherapy has recently emerged as a therapeutic option for patients with numerous posterior segment disorders. During the past decade, it has been used for the management of benign and malignant tumors, either as sole therapy or in combination with other treatments such a plaque radiotherapy. More recently, it has been used for management of choroidal neovascularization (CNV) secondary to age-related macular degeneration. A prospective, randomized, sham controlled multicenter clinical trial (TTT4CNV) is currently underway to evaluate the effectiveness of TTT for the management of CNV. PMID- 12777936 TI - Who should receive oral supplement treatment for age-related macular degeneration? AB - Oxidative damage to the retina has been proposed as a risk factor for age-related macular degeneration (AMD). Dietary or supplemental antioxidants may play a protective role. The Age-Related Eye Disease Study (AREDS), a randomized, multicenter, placebo-controlled clinical trial designed to test the effect of pharmacologic doses of antioxidants and zinc on the incidence and progression of AMD, reported a beneficial effect of high-dose supplements, taken for approximately 6 years, in delaying the progression of intermediate AMD to advanced AMD. AREDS and subsequent research on dietary intake or supplement use have not indicated a protective role of antioxidant or zinc intake or supplement use in the incidence or prevalence of early AMD. Numbers of cases were insufficient to investigate effects on late AMD. Persons with intermediate AMD and without contraindications should consider using antioxidant and zinc supplements. There is no evidence to date that earlier use conveys benefit. PMID- 12777937 TI - Update on photodynamic therapy. AB - To date, photodynamic therapy with verteporfin has been shown to benefit those patients with age-related molecular degeneration and choroidal neovascularization that is subfoveal and predominately classic (> 50%). As of this writing, the Food and Drug Administration is requiring additional data before verteporfin is approved for treatment of occult subfoveal choroidal neovascularization. Photodynamic therapy has also proved beneficial for subfoveal lesions secondary to high myopia. Although there is potential for patients with angioid streaks, ocular histoplasmosis syndrome, and idiopathic causes of choroidal neovascularization to benefit from photodynamic therapy, randomized clinical trials have not been performed. Photodynamic therapy has not been shown to benefit patients with minimally classic (< 50%) lesions. PMID- 12777938 TI - 2003 European Society of Hypertension-European Society of Cardiology guidelines for the management of arterial hypertension. PMID- 12777939 TI - Cardiovascular prevention and blood pressure reduction: a quantitative overview updated until 1 March 2003. AB - BACKGROUND: In a meta-analysis published in October 2001, we reported that new and old classes of antihypertensive drugs had similar long-term efficacy and safety. Furthermore, we observed that in clinical trials in hypertensive or high risk patients gradients in systolic pressure accounted for most differences in outcome. OBJECTIVE: To test whether our previous conclusions would hold, we updated our quantitative overview with new information from 14 clinical trials presented before 1 March 2003. METHODS: To compare new and old antihypertensive drugs, we computed pooled odds ratios from stratified 2 x 2 contingency tables. If Zelen's test of heterogeneity was significant, we used a random effects model. In a meta-regression analysis, we correlated odds ratios with corresponding between-group differences in systolic pressure. We then contrasted observed odds ratios with those predicted from gradients in systolic pressure. MAIN OUTCOMES: Differences in achieved systolic blood pressure and incidence of total and cardiovascular mortality, cardiovascular events, stroke, myocardial infarction and heart failure. NEW VERSUS OLD DRUGS: In 15 trials, 120 574 hypertensive patients were randomized to old drugs (diuretics or beta-blockers) or new agents [calcium-channel blockers, alpha-blockers, angiotensin-converting enzyme (ACE) inhibitors or angiotensin type-1 receptor (AR1) blockers]. Old and new drugs provided similar protection against total and cardiovascular mortality and fatal plus non-fatal myocardial infarction. Calcium-channel blockers, including (-8%, P = 0.07) or excluding verapamil (-10%, P = 0.02), as well as AR1 blockers (-24%, P = 0.0002) resulted in better stroke prevention than did the old drugs, whereas the opposite trend was observed for ACE inhibitors (+10%, P = 0.03). The risk of heart failure was higher (P < 0.0001) on calcium-channel blockers (+33%) and alpha-blockers (+102%) than on conventional therapy involving diuretics. META REGRESSION: Between-group differences in achieved systolic pressure ranged from 0.1 to 3.2 mmHg in seven actively controlled trials (73 237 patients), and from 2.1 to 22.1 mmHg in seven studies comparing varying intensities of blood pressure lowering (11 128 patients). For these 14 new trials, we predicted outcome from achieved systolic blood pressure using our previously published meta-regression models based on 30 trials with 149 407 patients. In general, predicted and observed odds ratios were similar. Larger reductions in systolic pressure (weighted mean 1.8 mmHg) in two trials accounted for the advantage of AR1 blockers over conventional therapy in the prevention of stroke. Only for cardiovascular mortality in very old patients (P = 0.02) and for cardiovascular events and myocardial infarction in old Australians (P < 0.05), the observed odds ratios deviated from our predictions based on the gradients in systolic blood pressure. INTERPRETATION: The hypothesis that new antihypertensive drugs, such as calcium-channel blockers, alpha-blockers, ACE inhibitors or AR1 blockers might influence cardiovascular prognosis over and beyond their antihypertensive effects remains unproven. The finding that blood pressure differences largely accounted for cardiovascular outcome emphasizes the desirability of tight blood pressure control. However, the level to which blood pressure must be lowered to achieve maximal benefit remains currently unknown. PMID- 12777940 TI - Genetic determinants of the dynamics and kinetics of alcohol as an environmental modifier of blood pressure. PMID- 12777941 TI - Anti-hypertensive and natriuretic effect of glucagon-like peptide 1 in Dahl S rats: a novel function for a pleotrophic hormone? PMID- 12777942 TI - Nitric oxide in diabetes mellitus. PMID- 12777943 TI - Adrenomedullin: a new member of the vasoactive peptide family with trophic action. PMID- 12777944 TI - Erythrocyte-derived depressing factor: a novel cardiovascular player? PMID- 12777945 TI - Risk stratification by guidelines compared with risk assessment by risk equations applied to a MONICA sample. AB - BACKGROUND: The World Health Organization/International Society of Hypertension (WHO/ISH) Hypertension Guidelines from 1999 propose a risk stratification scheme for estimating absolute risk for cardiovascular disease (CVD). Risk equations estimated by statistical methods are another way of predicting cardiovascular risk. OBJECTIVE: We studied the differences between these two approaches when applied to the same set of individuals with high blood pressure. DESIGN AND METHODS: The two northernmost counties in Sweden (NSW) constitute one of the centres in the WHO MONICA (monitoring trends and determinants in cardiovascular disease) Project. Three population surveys have been carried out in 1986, 1990 and 1994, and were used to estimate a risk equation for predicting the 10-year risk of fatal/non-fatal stroke and myocardial infarction. Another MONICA sample from 1999, a total of 5997 subjects, was classified according to the recent WHO/ISH risk stratification scheme. A risk assessment was also performed, by using the risk equations from the NSW MONICA sample and Framingham risk equations. RESULTS: The agreement between the two methods was good when the values obtained from the risk equation were averaged for each risk group obtained from the risk classification by guidelines. However, if the predicted risk for each individual was considered, the agreement was poor for the medium and high risk groups. Although the average risk for all individuals is the same, many subjects have a higher risk or a lower risk than predicted by guidelines. CONCLUSIONS: Risk classification by the 1999 WHO/ISH Hypertension Guidelines is not accurate and detailed enough for medium- and high-risk patients, which could be of clinical importance in the medium risk group. PMID- 12777946 TI - Do the ethanol metabolizing enzymes modify the relationship between alcohol consumption and blood pressure? AB - BACKGROUND: Several cross-sectional studies have examined whether the relationship between alcohol consumption and blood pressure (alcohol-BP relationship) differs among individuals with different aldehyde dehydrogenase-2 (ALDH2) genotypes, but few studies have examined the association with alcohol dehydrogenase-2 (ADH2), and those have yielded inconsistent results. We examined the potential modulatory effects of ADH2 and ALDH2 genotypes on the alcohol-BP relationship in a cross-sectional sample of a Japanese rural community. METHODS AND RESULTS: The study subjects were 335 randomly selected men aged 40-69 years, who lived in Shiso, a Japanese rural county, in 1999 or 2000. The genetic polymorphisms of ADH2 and ALDH2 were determined by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) methods. The frequencies of ADH21/21 (wild-type), 21/22 (superactive heterozygotes), and 22/22 (superactive homozygotes) were 8.4, 34.9 and 56.7%, respectively; and those of ALDH21/21 (wild type), 21/22 (inactive heterozygotes), and 22/22 (inactive homozygotes) were 52.8, 40.9, and 6.3%, respectively. A multiple linear regression analysis showed that the relationship between alcohol consumption and diastolic blood pressure was significantly stronger in men with ADH21/21 than those with ADH21/22 or 22/22 (adjusted regression coefficient = 0.0392 versus 0.0113 mmHg for + 1 g ethanol/week, P for difference in slope = 0.018). The strength of the alcohol-BP relationship was similar in all of the ALDH2 genotype groups. CONCLUSION: The alcohol-BP relationship was significantly stronger in men with ADH21/21 than in men with ADH21/22 or 22/22 in this Japanese rural population. This finding was exactly the opposite of what one previous study suggested. PMID- 12777947 TI - Circadian gene expression of clock genes and plasminogen activator inhibitor-1 in heart and aorta of spontaneously hypertensive and Wistar-Kyoto rats. AB - OBJECTIVE: Heart and aorta possess biologic clocks, but their involvement in genetic hypertension has been unknown. Plasminogen activator inhibitor-1 (PAI-1) expression is directly regulated by clock genes, while angiotensin II modulates both PAI-1 and clock gene expression. We therefore examined circadian expression of PAI-1 and clock genes, and effects of angiotensin type 1 (AT1) receptor antagonism, in heart and aorta of spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) rats. METHODS: We examined cardiac and aortic mRNA expression for PAI-1 and clock genes (Per2, Bmal1, Clock, and Dbp) every 4 h throughout the day by quantitative reverse transcription-polymerase chain reaction, and intervention with the AT1 receptor antagonist candesartan and equihypotensive hydralazine. RESULTS: Cardiac PAI-1 expression was high in the dark, while aortic PAI-1 expression was high in the light. Both cardiac and aortic PAI-1 expression were greater in SHR than in WKY rats. Candesartan treatment decreased cardiac PAI 1 expression only in the dark in WKY rats but throughout the day in SHR. Candesartan but not hydralazine strongly attenuated circadian fluctuation of aortic PAI-1 mRNA in SHR and WKY rats. Clock genes oscillated synchronously in heart and aorta of SHR and WKY rats. Clock gene expression was increased in heart but not aorta of SHR. Candesartan did not affect clock gene expression. CONCLUSIONS: Enhanced expression of clock genes may increase PAI-1 expression in concert with activated renin-angiotensin system in SHR heart. Rather than clock genes, the renin-angiotensin system induces daily fluctuation and increased expression of aortic PAI-1 mRNA in SHR. PMID- 12777948 TI - Age-related alterations in soluble guanylyl cyclase and cGMP pathway in spontaneously hypertensive rats. AB - BACKGROUND: Vascular contractility and blood pressure (BP) are regulated by soluble guanylyl cyclase (sGC) and cyclic guanosine monophosphate (cGMP) pathway, which can be influenced by heme oxygenase (HO)-derived carbon monoxide (CO). The age-related changes in sGC/cGMP pathway in tail artery smooth muscle cells (SMCs) in hypertension have not been systematically investigated. METHODS: In the present study, spontaneously hypertensive rats (SHR) of 4, 8, and 20 weeks old were used. The basal and hemin-modulated levels of sGC and cGMP in tail artery tissues were examined. RESULTS: Although BP of 20-week SHR was significantly elevated, sGC and cGMP levels were unaltered compared with age-matched Wistar Kyoto rats (WKY). The levels of sGC and cGMP were significantly lower in 4- and 8 week SHR compared with age-matched WKY although BP of 4-week SHR was normotensive. Hemin administration resulted in a significant decrease in BP in 8 week (158.7 +/- 2.4 versus 123.5 +/- 1.3 mmHg, P < 0.01), but not in pre hypertensive (4 weeks) or 20-week SHR or WKY at all ages. Coincidently, sGC and cGMP levels in 8-week SHRs were significantly elevated and so did the expression levels of HO-1. Hemin treatment did not increase the cyclic adenosine monophosphate (cAMP) content of tail artery from 8-week SHR. The constitutive HO 2 levels remained unchanged in 8- and 20-week SHR and age-matched WKY. CONCLUSION: The HO-activity inhibitor, chromium mesoporphyrin, abolished the BP lowering and HO- stimulating effects of hemin in young SHR. Our results suggest that alteration in sGC/cGMP pathway in vascular SMCs precedes the occurrence of hypertension but returns to normal once hypertension is fully manifested. PMID- 12777949 TI - Antihypertensive effect of glucagon-like peptide 1 in Dahl salt-sensitive rats. AB - BACKGROUND: Dahl salt-sensitive (Dahl S) rats exhibit many phenotypic traits associated with salt-sensitive hypertension in man. Specifically, they are salt sensitive, insulin-resistant and hyperlipidemic. They also develop endothelial dysfunction, cardiac injury and glomerulosclerosis. Insulin resistance is linked to hypertension, renal and cardiac damage and endothelial dysfunction. Thus, an agent that has diuretic action and can improve insulin resistance, like recombinant glucagon-like peptide-1(7-36)amide (rGLP-1), may have an antihypertensive effect. OBJECTIVE: To determine whether chronic administration of rGLP-1 attenuates the development of hypertension, endothelial dysfunction and/or hypertension-induced renal and cardiac end organ damage in Dahl S rats. METHODS: Mean arterial pressure (MAP) and urinary excretion of protein and albumin were measured in Dahl S rats before and after they were fed a 8% NaCl diet and infused with rGLP-1 (1 micro g/kg per min, i.v.) or vehicle for 14 days. At the end of the study, the degree of renal and cardiac injury was histologically assessed and endothelium-dependent relaxing function was studied using aortic rings. In other rats, the effects of rGLP-1 on sodium and water balance and plasma glucose and insulin levels for the first 3 days following a step change in sodium intake from a 0.1% NaCl diet to 7.5 mEq/day were determined. RESULTS: rGLP-1 significantly attenuated the development of hypertension in Dahl S rats (136 +/- 7 versus 174 +/- 6 mmHg). This was associated with reduction in proteinuria (46 +/- 7 versus 128 +/- 15 mg/day) and albuminuria (46 +/- 7 versus 86 +/- 18 mg/day) and improvement of endothelial function and renal and cardiac damage. rGLP-1 markedly increased urine flow and sodium excretion for the first 3 days following elevation in sodium intake. It had no significant effects on plasma glucose and insulin concentrations. CONCLUSION: rGLP-1 has antihypertensive and cardiac and renoprotective effects in Dahl S rats fed a high salt diet. The antihypertensive effect of rGLP-1 in Dahl S rats is due mainly to its diuretic and natriuretic effects, rather than an effect to improve insulin-resistance. PMID- 12777951 TI - Progesterone does not influence vascular function in postmenopausal women. AB - BACKGROUND: The protective functions of oestrogen therapy alone on cardiovascular risk parameters are well established; however, the action of progesterone on vascular parameters in an oestrogen-deprived environment is less clear. OBJECTIVES: To examine the effects of progesterone alone on vascular function and hormone levels in postmenopausal women. DESIGN: In a randomized, double-blind, cross-over design study, 20 healthy postmenopausal women were tested before and after 6 weeks of treatment with micronized progesterone (100 mg/daily) and matching placebo. METHODS: Tests included measurement of sex hormones and gonadatropin levels, lipids and measures of surrogate markers of vascular function including, blood pressure, flow-mediated dilation of the brachial artery, systemic arterial compliance and cutaneous vascular reactivity. RESULTS: The mean (+/- SEM) age of subjects was 56.4 +/- 2.7 years and the average body mass index at the baseline visit was 27.1 +/- 1.0 kg/m2. Progesterone levels increased as a result of progesterone treatment (0.9 +/- 0.2 to 9.5 +/- 2.3 nmol/l, P = 0.001), whereas follicle-stimulating hormone levels decreased (75.1 +/- 11.4 to 67.6 +/- 10.0, P = 0.001). Systemic arterial compliance, flow mediated dilation, cutaneous vascular reactivity, blood pressure, body mass index, plasma levels of cholesterol, lipids and oestrogen were unchanged. CONCLUSIONS: We conclude that progesterone given without oestrogen does not adversely affect vascular function in postmenopausal women. PMID- 12777950 TI - Effect of glycaemic control on the vascular nitric oxide system in patients with type 1 diabetes. AB - OBJECTIVE: The mechanisms underlying the relationship between the vascular complications of diabetes and the glycaemic control are not well understood. We tested whether glycaemic control influences the functioning of the nitric oxide system in type 1 diabetic patients and the role for oxidative stress. METHODS: The changes in the forearm blood flow after the infusion in the brachial artery of NG-monomethyl-l-arginine, methacholine, methacholine plus superoxide dismutase, and nitroprusside were evaluated using strain gauge plethysmography in 14 healthy subjects and 24 patients with type 1 diabetes (12 with HbA(1c) < 7.5%; 12 with HbA(1c) > or = 7.5%). After adjusting insulin treatment, the vascular studies were repeated in the initially poorly controlled patients (HbA(1c) > or = 7.5%). RESULTS: Compared with healthy people, impaired vascular responses to NG monomethyl-l-arginine (P = 0.0001), methacholine (P = 0.007) and nitroprusside (P = 0.0015) were found in the patients with type 1 diabetes and a poor glycaemic control (HbA(1c) >/= 7.5%), but not in subjects with good control (HbA(1c) < 7.5%). Superoxide dismutase improved the responses to methacholine only in those patients with poor control (P = 0.0037). After the adjustment of the insulin treatment in poorly-controlled patients, the responses improved and the effect of superoxide dismutase disappeared only in the patients that achieved good control (n = 9), but not in those who remained poorly-controlled (n = 3). CONCLUSIONS: In patients with diabetes type 1, glycaemic control determines the functioning of the NO system by a reversible mechanism involving superoxide anions. This finding provides an explanation of the relationship between glycaemic control and vascular complications in diabetes. PMID- 12777952 TI - Hormone replacement therapy causes a decrease in hepatocyte growth factor in hypertensive women. AB - OBJECTIVE: Serum hepatocyte growth factor (HGF) is associated with blood pressure. We investigated whether the serum HGF level differs between hypertensive and normotensive postmenopausal women (PMW) and whether hormone replacement therapy (HRT) alters the serum HGF level and blood pressure in hypertensive and normotensive PMW. DESIGN: Prospective observational study. METHODS: A total of 33 PMW with mild to moderate essential hypertension controlled by antihypertensive treatment (mean age, 57 +/- 6 years) and 23 normotensive PMW (mean age, 57 +/- 7 years) received continuous HRT (0.625 mg of conjugated equine estrogen combined with 2.5 mg of medroxyprogesterone acetate) once a day orally for 12 months, and we measured serum HGF levels and blood pressure before and 12 months after the start of HRT. RESULTS: The baseline serum HGF level was significantly higher in hypertensive PMW than in normotensive PMW. HRT significantly decreased the serum HGF level in hypertensive subjects, from 2.85 +/- 0.64 pmol/l to 2.49 +/- 0.65 pmol/l (P < 0.001), but not in normotensive subjects. HRT did not change blood pressure in either group. CONCLUSIONS: Serum HGF level before the start of HRT was higher in the hypertensive PMW than in the normotensive PMW. Furthermore, HRT decreases serum HGF without decreasing blood pressure in hypertensive PMW. The HRT-induced decrease in serum HGF was greater in hypertensive PMW than in normotensive PMW, and the decrease was independent of blood pressure changes. PMID- 12777953 TI - Prevalence of renal artery stenosis in high-risk veterans referred to cardiac catheterization. AB - OBJECTIVE: The prevalence of atherosclerotic renal artery stenosis (RAS) varies depending on patient selection with no specific guidelines on indications for selective renal angiography in patients referred for coronary angiography. The goal of this study is to determine the prevalence and predictors of renal artery stenosis in hypertensive veterans referred for coronary angiography. DESIGN: Prospective study. SETTING: Tertiary care veterans' administration facility in the USA. PATIENTS, PARTICIPANTS: A total of 90 veterans referred for coronary angiography with an initial ascending aortic pressure > 135 mmHg. INTERVENTIONS: Selective renal angiography was performed following coronary angiography. RESULTS: We found that 28% of the patients had single RAS (> or = 50% stenosis), while 16% had single RAS > or = 70% stenosis, 10% had bilateral RAS >or = 50% and 6% had bilateral RAS > or = 70%. Significant positive univariate predictors of RAS (> or = 50%) were age, peripheral vascular disease (PVD), creatinine level (Cr) and myocardial infarction. Significant multivariate predictors of RAS (> or = 50%) were age > 65 years [relative risk (RR), 3.6; 95% confidence interval (CI), (1.2-10.6)], PVD [RR 3.2, 95% CI (1.1-9.1)] and Cr > 1 mg/dl [RR 4.9, 95% CI (1.53-15.9)]. No complications related to renal angiography were noted. CONCLUSIONS: Selective renal angiography during routine coronary angiography in hypertensive veterans with coronary artery disease is safe and uncovers RAS in many older patients with PVD and renal insufficiency. PMID- 12777954 TI - Microalbuminuria, a parameter independent of metabolic influences in hypertensive men. AB - OBJECTIVE: To evaluate the relationship of albuminuria and microalbuminuria (overnight urine albumin > or = 15 micro g/min) with insulin resistance and related metabolic abnormalities in patients with essential hypertension. DESIGN: Cross-sectional evaluation of 271 (age range, 19-77 years) never-treated, non diabetic, uncomplicated hypertensive men. MAIN OUTCOME MEASURES: Triplicate overnight urine albumin determination and homeostasis model assessment (HOMA) of insulin resistance as a surrogate measure of insulin sensitivity. Additional parameters were fasting and post-load circulating glucose and insulin, lipids, body mass index, blood pressure and echocardiographic left ventricular mass. RESULTS: HOMA, fasting and post-challenge glucose and insulin, percentages of glucose-intolerant patients, triglycerides and high-density lipoprotein cholesterol levels did not differ across ascending urine albumin quartiles. Body mass index, blood pressure and ventricular mass were significantly greater in the upper quartiles, and the prevalence of obesity fivefold more frequent in the top as compared with the bottom urine albumin fourth. The statistical trend was unchanged after adjustment for HOMA, while accounting for systolic blood pressure and left ventricular mass by co-variance analysis abolished it. Eighty-eight patients bearing the phenotypic traits of the metabolic syndrome and a striking degree of insulin resistance and hyperinsulinemia showed urine albumin rates and prevalence of microalbuminuria comparable with the 183 patients who were not affected by that syndrome. CONCLUSIONS: Albuminuria is independent of insulin resistance and other phenotypic components of the metabolic syndrome in never treated, non-diabetic essential hypertensive men. Microalbuminuria is more frequent in obese hypertensives but this association is explained by higher blood pressure more than insulin resistance. PMID- 12777955 TI - Upregulation of ligand, receptor system, and amidating activity of adrenomedullin in left ventricular hypertrophy of severely hypertensive rats: effects of angiotensin-converting enzyme inhibitors and diuretic. AB - OBJECT: We investigated the pathophysiological role of the cardiac adrenomedullin (AM) system, including the ligand, receptor and amidating activity in the hypertrophied heart in severe hypertension. METHOD: We studied the following four groups: control Wistar-Kyoto rats (WKY), spontaneously hypertensive stroke-prone rats (SHR-SP), 8 weeks captopril-treated SHR-SP, and 8 weeks trichlormethiazide treated SHR-SP. AM precursor is converted to inactive glycine-extended AM (AM Gly) and subsequently AM-Gly is converted to active mature AM (AM-m) by enzymatic amidation. We measured AM-m, AM-total (AM-T; AM-T = AM-m + AM-Gly), and atrial natriuretic peptide (ANP) in the plasma and left ventricle (LV) by immunoradiometric assay. We also measured gene expression of AM and ANP was and gene expression and protein levels of AM receptor system components such as calcitonin receptor-like receptor (CRLR), receptor-activity modifying protein (RAMP) 2 and RAMP3. RESULTS: At 7 weeks old, SHR-SP had higher blood pressure and ANP mRNA levels and lower plasma AM-T compared with WKY, however, there were no differences in other indices between the two groups. At 17 weeks old, SHR-SP had increased blood pressure, LV weight, plasma and LV ANP levels and mRNA levels of ANP compared with WKY. AM-m and AM-T levels in plasma (AM-m: + 31%; AM-T: + 56%) and the LV (AM-m: + 84%; AM-T: + 31%) were significantly higher in SHR-SP than in WKY. The LV tissue AM-m/AM-T ratio was significantly higher in SHR-SP (93.2%) than in WKY. The mRNA levels of AM, CRLR, and RAMP2 in the LV were significantly higher in SHR-SP than in WKY. Captopril and trichlormethiazide similarly decreased blood pressure and LV hypertrophy with the reduction of the LV AM-m and AM-T levels and mRNA abundance of AM and its receptor component. CONCLUSION: These results suggest that cardiac AM system is upregulated in the hypertrophied heart in this hypertension model. Considering that AM acts as an anti-remodeling autocrine and/or paracrine factor, upregulation of the AM system may modulate the pathophysiology in LV hypertrophy. PMID- 12777957 TI - Awareness, treatment and control of hypertension in patients attending hospital clinics in China. AB - OBJECTIVES: To determine awareness, treatment and control of hypertension in patients attending hospital clinics in China. DESIGN: A cross-sectional survey. PATIENTS AND SETTING: Patients over the age of 35 years, who were attending outpatient clinics in 18 hospitals of eight major cities of Northern and Southern China, were interviewed face-to-face between June and July 1999. METHODS: Trained fieldworkers completed questionnaires regarding demographics, hypertension knowledge and awareness, treatment history and quality of life issues. Qualified physicians performed blood pressure assessment. RESULTS: A total of 9703 volunteers enrolled, of which 4510 (46.5%) were hypertensives. Among hypertensives, 23% were unaware of their high blood pressure. Although 89% of those aware reported receiving therapy, only 56% of them were taking medication regularly, and 33% were thus controlled. Although 69% of all hypertensives measuring at examination reported taking antihypertensive therapy, only 44% of them were actually taking medication regularly. Furthermore, 73% of hypertensives believed hypertension was not a serious problem, and required no regular medication. CONCLUSIONS: The results showed that a substantial number of outpatients have a hypertensive range of blood pressure. Although most of these patients are already being treated with drug regimen, the patient's misconceptions and lack of knowledge about the disease appear to be the major cause of treatment failure. The low rate of high blood pressure control for both sexes calls for further improvements. PMID- 12777956 TI - Influence of hypertension on cardiac contractile response of human erythrocyte derived depressing factor in ventricular myocytes. AB - BACKGROUND: Erythrocyte-derived depressing factor (EDDF), a novel hypotensive factor purified from human erythrocytes, elicits endothelium-dependent vasorelaxation by reducing intracellular Ca2+ in vascular smooth muscle cells. However, its cardiac response is unknown. OBJECTIVE: This study was designed to examine the cardiac contractile response of EDDF under both normotensive and hypertensive conditions. METHODS: Ventricular myocytes were isolated from adult male spontaneously hypertensive rats (SHR) and age-matched Wistar-Kyoto (WKY) normotensive rats. Mechanical properties were evaluated using an IonOptix MyoCam system and intracellular Ca2+ was measured with fura-2 fluorescence. Myocytes were electrically stimulated to contract at 0.5 Hz. The contractile properties analyzed included peak shortening (PS), time-to-PS (TPS), time-to-90% re lengthening (TR(90)), maximal velocity of shortening/re-lengthening (+/- dl/dt), fura-fluorescence intensity change (DeltaFFI), and fura-fluorescence decay rate (tau). RESULTS: SHR rats displayed significantly elevated blood pressure. EDDF (10-9-10-4 g/ml) did not affect PS, TPS, TR(90), DeltaFFI and tau but depressed +/- dl/dt at higher doses in WKY myocytes. However, EDDF depressed PS, +/- dl/dt and DeltaFFI, shortened TPS without affecting TR(90) and tau in SHR myocytes. Pretreatment of the myocytes with the nitric oxide synthase inhibitor Nvarpi nitro-l-arginine methyl ester (l-NAME) did not affect the EDDF-induced inhibition of PS and +/- dl/dt in SHR myocytes but unmasked an EDDF-induced negative response in WKY myocytes. CONCLUSIONS: These data indicate that EDDF may participate in the modulation of cardiac contractile function under hypertensive, but not normotensive, conditions. The cardiac depressive effect of EDDF is unlikely due to release of nitric oxide, as suggested in vascular smooth muscles. PMID- 12777958 TI - Why is hypertension so frequently uncontrolled in secondary prevention? AB - OBJECTIVE: To analyze blood pressure (BP) control in secondary prevention. DESIGN: Individual data of two cross-sectional studies on preventive cardiology (PRATIK and ESPOIR studies conducted, respectively, in general practice and with private cardiologists) were analyzed. SETTING: Primary care. PARTICIPANTS: Patients both with treated hypertension and coronary disease. MAIN OUTCOME MEASURES: Risk factors, treatments, cardiovascular history and BP were recorded. Each population was divided in three groups: group I, no other risk factor; group II, one or two risk factors; group III, three or more risk factors or diabetes. RESULTS: A total of 1423 and 2596 patients, respectively, recruited in general practice and by cardiologists were analyzed. Of these, 473 (33.24%) and 1060 (40.83%) patients, respectively, had controlled hypertension. Among uncontrolled hypertensives, more than 50% had borderline isolated systolic hypertension. Associated risk factors negatively affect hypertension control, which had been achieved in a lower percentage of patients in group III than in group I (general practice, 26.28 versus 42.20%; cardiological practice, 32.42 versus 56.13%). In general practice, the percentage of patients receiving beta-blockers was significantly lower in group III. Among individuals with uncontrolled hypertension, only 17.58 and 26.69% received at least three-drug treatment including diuretics in general and in cardiological practice, respectively. CONCLUSION: The negative influence of associated risk factors and the under-use of combination therapy contribute to poor BP control. In addition the high frequency of borderline isolated systolic hypertension suggests that the prerequisite to improve hypertension control should be to convince practitioners of the beneficial effect of tight systolic BP control (below 140 mmHg) in secondary prevention. PMID- 12777960 TI - In search of endogenous CYP2D6 substrates. PMID- 12777959 TI - Effects of antihypertensive agents, alpha receptor blockers, beta blockers, angiotensin-converting enzyme inhibitors, angiotensin receptor blockers and calcium channel blockers, on oxidative stress. AB - BACKGROUND: Free oxygen radicals and insufficiency of antioxidant enzymes have been implicated in the pathogenesis of hypertension disease (HD). Trace elements function as a co-factor in antioxidant enzymes. The antioxidant system and trace elements have been investigated in many different studies including HD, but these subjects have not been investigated as a whole in these patients. OBJECTIVE: The aim of the present study was to investigate the antioxidative system and trace elements in hypertensive patients given different antihypertensive therapy. METHODS: We examined malondialdehyde and superoxide dismutase activities together with copper and zinc levels in plasma of 102 patients with HD and in 51 healthy controls. RESULTS: It was found that in patients with HD, plasma malondialdehyde was significantly higher than those of controls, while plasma superoxide dismutase activities were significantly lower in patients with HD. Plasma zinc levels were significantly higher than those of controls and plasma copper levels were significantly lower in patients with HD. Plasma lipid levels and oxidative state were analyzed in five different treatment groups given antihypertensive drug therapy before and after a 3-month treatment period. CONCLUSIONS: In conclusion, our clinical study shows that angiotensin-converting enzyme inhibitors and angiotensin receptor blockers have notable effects on oxidative stress, and are an essential step in managing essential hypertension by the way of improvement of endothelial dysfunction. Although it has been shown that calcium channel blockers, beta blockers and alpha receptor blockers have antioxidant effects in in vitro conditions, we did not demonstrate these effects in our clinical study. PMID- 12777961 TI - Screening for endogenous substrates reveals that CYP2D6 is a 5 methoxyindolethylamine O-demethylase. AB - The objective of this investigation was to screen for potential endogenous substrates for CYP2D6. Using recombinant CYP2D6, together with hepatic microsomes from CYP2D6-transgenic mice, human liver microsomes, and a specific anti-CYP2D6 monoclonal antibody, it was ascertained that CYP2D6 does not significantly metabolize the endogenous phenylethylamines 2-phenylethylamine, octopamine, synephrine, 3-methoxy-p-tyramine, 4-methoxy-m-tyramine, metanephrine, and normetanephrine, nor the indolethylamines tryptamine, serotonin, 6 methoxytryptamine, and melatonin, nor the beta-carbolines harman, norharman and tryptoline. However, the indolethylamines 5-methoxy-N,N-dimethyltryptamine (5 MDMT) and pinoline (6-methoxy-1,2,3,4-tetrahydro-beta-carboline) showed relatively high affinity for CYP2D6 in a spectral binding assay (K(s) 28 +/- 5, and 0.5 +/- 0.3 microm (mean +/- SEM), respectively) and were O-demethylated only by CYP2D6 in a panel of 15 recombinant common human P450s. Pinoline and 5-MDMT O demethylase activities were 35- and 11-fold greater in liver microsomes from CYP2D6-humanized mice, respectively, than those in liver microsomes from control mice. Moreover, the increased activities were completely inhibited by an anti CYP2D6 monoclonal antibody. Kinetic analysis with recombinant CYP2D6 gave K(m) and k(cat) values for 5-MDMT and pinoline O-demethylations of 12 +/- 1 microm and 65 +/- 1 min(-1) and 1.8 +/- 0.3 microm and 26 +/- 1 min(-1), respectively. These two substrates can be added to 5-methoxytryptamine, which we have recently reported to be an endogenous CYP2D6 substrate. CYP2D6 is therefore a relatively highly specific, high-affinity, high-capacity 5-methoxyindolethylamine O demethylase. Polymorphic cytochrome CYP2D6 may therefore exert an influence on mood and behavior by the O-demethylation of these 5-methoxyindolethylamines found in the brain and pineal gland. These processes may also impact on mental and neurological health. The findings may open new vistas for the determination of CYP2D6 phenotype. PMID- 12777963 TI - Confirmation of quantitative trait loci for cocaine-induced activation in the AcB/BcA series of recombinant congenic strains. AB - Individual differences in the psychomotor stimulant effects of cocaine are influenced by genetic factors. Several quantitative trait loci (QTL) have been identified for cocaine-induced locomotor activation using the AXB/BXA recombinant inbred series of strains derived from the A/J (A) and C57BL/6J (B6). The aim of the present study was to conduct an independent analysis of cocaine-induced activation in the AcB/BcA recombinant congenic strains. The AcB/BcA RC series consists of 37 inbred strains derived from reciprocal backcrosses between the A and B6, followed by systematic inbreeding. Locomotor activity was measured in a computerized open-field apparatus following intraperitoneal administration of saline and cocaine (20 mg/kg). Linkage maps constructed with 625 informative microsatellite markers were used to identify chromosomal regions associated with cocaine difference scores. Significant (P < 0.00001) regions were identified on chromosomes 1 (13-25.7 and 36.9-58.5 cM), 5 (1-28 and 84-86 cM), 6 (7-26.35 cM), 7 (9.4-27.8 cM), 9 (9-28 cM), 13 (21-37 cM), 16 (36-66 cM), 17 (22.5-24.5 cM) and 18 (45-48 cM). Multiple regression analysis demonstrated that a subset of four markers, including D5Mit182 (24 cM), D5Mit409 (84 cM), D7Mit83 (26.5 cM) and D13Mit54 (35 cM), accounted for 90% of the genetic variance in cocaine difference scores. The results of the present study provide confirmation for a number of QTL on chromosomes 1, 5, 6, 9, 16 and 17 which were previously identified in the recombinant inbred AXB/BXA and BXD strains that share a common B6 ancestor. PMID- 12777962 TI - CYP2E1*1D regulatory polymorphism: association with alcohol and nicotine dependence. AB - OBJECTIVE: CYP2E1 bioactivates environmental protoxins and metabolizes alcohol. CYP2E1 is induced by alcohol and cigarette smoking and may contribute to metabolic tolerance in alcoholics. The CYP2E1*1D polymorphism has been associated with greater CYP2E1 inducibility. One objective was to determine the frequency of the variant allele in eight ethnic groups. Further, the Canadian Native Indian, South-east Asian Canadian and Caucasian Canadian groups were stratified by alcohol and nicotine dependence (as measured by DSM-IV criteria) to examine the potential association of CYP2E1*1D with drug dependence. RESULTS AND CONCLUSIONS: We found a significantly greater frequency of the CYP2E1*1D allele among Indo Asian Canadians (0.31), Chinese Canadians (0.19), Taiwanese (0.20), Japanese Canadians (0.18), African Americans (0.13), African Canadians (0.10) and Canadian Native Indians (0.09) compared to Caucasian Canadians (0.02). Although the power of the association study was low among some subgroups, the CYP2E1*1D genotype (subjects with at least one variant allele) was associated with alcohol as well as nicotine dependence. Specifically, Canadian Native Indians dependent on nicotine alone or alcohol alone exhibited significantly greater CYP2E1*1D frequencies compared to non-drug dependent controls, while the variant frequency among Southeast Asians dependent on nicotine was greater than their non-drug dependent counterparts. We also found that CYP2E1*1D genotype was associated with significantly greater 3-hydroxycotinine per cigarette in African Americans. The variable frequency of CYP2E1*1D among ethnic groups suggests a greater risk for diseases putatively related to CYP2E1 in some non-Caucasian ethnic groups. The association of CYP2E1*1D with alcohol and nicotine dependence suggests that CYP2E1 may contribute to the development of these dependencies. PMID- 12777964 TI - Structure and polymorphisms of human aryl hydrocarbon receptor repressor (AhRR) gene in a French population: relationship with CYP1A1 inducibility and lung cancer. AB - OBJECTIVE: The aryl hydrocarbon receptor repressor (AhRR) protein may dimerize with the AhR nuclear translocator (ARNT) and may compete with the aryl hydrocarbon receptor (AhR) to bind the xenobiotic responsive elements. The result is a negative feedback mechanism that involves a down regulation of all genes regulated by the AhR transcription factor which positively regulates the expression of the Cytochrome P-4501A1 gene (CYP1A1). METHODS: The structure of the AhRR gene was reconstituted, then the genetic polymorphisms of this gene including the promoter were investigated and the link between these polymorphisms, CYP1A1 inducibility and lung cancer incidence in a French population was examined. Four polymorphisms were found, two in the coding region (609G>C and 1977G>C) and two in the 5'-untranslated region (-96G>A and -869A>T). Among the four polymorphisms, only one, the 609G>C has been previously described. The 609G>C and 1977G>C are localized respectively in exon 6 and 12 and lead to Pro554Ala and Asp641His substitutions, respectively. To evaluate the frequency of these allelic variants, a DNA library of a case-control study of lung cancer (164 controls and 171 patients) was screened. These polymorphisms were detected at the same allele frequency (0.40 for 609C, 0.05 for 1977C, 0.24 for -96A and 0.17 for 869T) in both controls and patients. Statistical analysis did not show any relationship between all the mutations found and CYP1A1 inducibility and lung cancer incidence. CONCLUSION: None of the polymorphisms were found to play a key role in CYP1A1 inducibility or in the susceptibility to develop lung cancer. PMID- 12777965 TI - CYP2E1 and NQO1 genotypes, smoking and bladder cancer. AB - BACKGROUND: Cytochrome P450 2E1 (CYP2E1) and NAD(P)H:quinone oxidoreductase (NQO1) catalyze the activation of some environmental procarcinogens present in tobacco smoke (i.e. nitrosoamines and heterocyclic amines). We conducted a hospital based case-control study to evaluate the potential association between genetic polymorphisms of CYP2E1 (C1019T in the 5' flanking region) and NQO1 (C609T in exon 6) and bladder cancer risk in Asian population. METHODS: The study population was comprised of 218 histologically confirmed prevalent bladder cancer cases and 199 controls without cancer or systemic illness. PCR-restriction fragment length polymorphism based methods were used for the genotyping analyses and unconditional logistic regression model for the statistical evaluations. RESULTS: The risk of bladder cancer increased with the amount of smoking (P for trend < 0.01). The frequency of CYP2E1 c1/c1 genotype was significantly higher in bladder cancer patients (57.9%) than in the controls (47.9%) (OR = 1.8, 95% CI = 1.1-2.9). Similarly, the NQO1 C/C genotypes were significantly more prevalent in the patients (45.8%) than in the controls (37.6%) (OR = 1.6, 95% CI = 1.0-2.7). The risk for bladder cancer increased with the number of the putative risk genotypes (P for trend = 0.03); the most remarkable risk was observed for heavy smokers with both CYP2E1 c1/c1 and NQO1 C/C genotypes (OR = 13.8, 95% CI = 3.9 48.6) when compared to non/light smokers with other genotypes. CONCLUSION: Our findings suggest that CYP2E1 and NQO1 genotypes may play an important role in development of smoking related bladder cancer among Korean men. PMID- 12777966 TI - Expression of human paraoxonase (PON1) during development. AB - BACKGROUND: Paraoxonase (PON1), a HDL-associated enzyme, protects against toxicity from specific organophosphorus compounds and oxidized lipids. Common polymorphisms in the PON1 gene have been identified and characterized in the coding region, 5' regulatory region and 3' UTR. The Q192R coding region polymorphism determines substrate-dependent differences in catalytic efficiency of hydrolysis. The -108CT polymorphism in the 5' regulatory region has a significant effect on PON1 expression, with the -108C allele expressing on average twice the level of plasma PON1 as the -108T allele. In addition to the effects of regulatory and coding region polymorphisms on PON1 levels and activity, plasma PON1 levels are also developmentally regulated. Since PON1 levels are important in determining resistance to specific organophosphorus compounds, the time course of appearance of PON1 in newborns is of great interest. RESULTS: We report here that PON1 levels plateau between 6 to 15 months of age, and that variability in the age at which PON1 levels plateau is quite variable among individuals. In mice and rats, plasma PON1 activity reaches a plateau at 3 weeks of age. In mice that lack endogenous PON1, human transgenes encoding either PON1(Q192) or PON1(R192) under the control of the human PON1 regulatory sequences exhibited a similar time course of expression as that seen in wild-type mice, indicating conservation of the developmental regulatory elements between mouse and human PON1. PMID- 12777968 TI - Thiopurine methyltransferase phenotypes and genotypes in Brazilians. AB - The polymorphism of thiopurine methyltransferase (TPMT) was studied in 306 healthy Brazilians who were classed, on the basis of self-declared colour and ancestry, as Euro-derived (n = 81), Afro-derived (n = 18) or having interethnic admixture (n = 204). TPMT activity (range 0.17-25.93 U) displayed a trimodal distribution of high (> 11.3 U; 9% of individuals), intermediate (5-11.3 U; 9.8%) and low (0.17 U; 0.3%) phenotypes. The occurrence of the TPMT mutations 238G>C, 460G>A and 719A>G was investigated in all individuals with low or intermediate phenotype, and in 43 with high-activity phenotype. None and two mutant alleles were associated with high- or low-activity phenotypes, respectively, whereas one mutant allele was detected in 26 of the 30 intermediate phenotype individuals. The allele frequencies of TPMT*2, TPMT*3A and TPMT*3C did not differ between individuals classed as Euro-derived (0.76%, 2.03% and 2.54%, respectively) or having interethnic admixture (0.60%, 1.81% and 1.81%, respectively). Furthermore, within each of these groups, the frequencies of TPMT*3A and TPMT*3C were not significantly different. PMID- 12777967 TI - Cholecystokinin, cholecystokinin-A receptor and cholecystokinin-B receptor gene polymorphisms in Parkinson's disease. AB - Cholecystokinin modulates the release of dopamine and dopamine-related behaviours in the mesolimbic pathway, where cholecystokinin and dopamine coexist in dopaminergic neurones. Because cholecystokinin and its receptors (A and B) have a functional interaction with dopaminergic neurotransmission, alterations in them may constitute a predisposition for Parkinson's disease. We performed a case control study to investigate the association between the cholecystokinin system and Parkinson's disease using genetic markers for three genes: cholecystokinin and its two receptors (A and B). One hundred and sixty patients with Parkinson's disease and 160 controls, matched for age, gender, ethnic origin and area of residence, were recruited. Cholecystokinin -45C>T, cholecystokinin-A receptor 779T>C and cholecystokinin-B receptor 1550G>A gene polymorphisms were studied using polymerase chain reaction-restriction fragment length polymorphism analyses. These three gene polymorphisms showed no correlation with risk of Parkinson's disease; however, the cholecystokinin CT/TT genotype was associated with a 4.429-fold increased risk for visual hallucinations in Parkinson's disease. Cholecystokinin-A receptor and B receptor polymorphisms, considered alone, showed no correlation with hallucinations in Parkinson's disease; however, a combined effect was found in patients with hallucinations harboring both the cholecystokinin CT/TT and cholecystokinin-A receptor TC/CC genotypes. Parkinson's disease patients harboring this genotype have a 5.922-fold increased risk for developing visual hallucinations. These results suggest that, in Chinese, visual hallucinations in Parkinson's disease are associated with cholecystokinin -45C>T polymorphism, and this association was still observed in the presence of the cholecystokinin-A receptor TC/CC genotype, indicating a possible interaction of these two genes in the visual hallucinogenesis in Parkinson's disease. PMID- 12777969 TI - Standards of practice for case managers: the bottom line. PMID- 12777970 TI - Rural case management for stroke: the development of a community-based screening and education program. AB - This article describes the evolution of a Community-Based Stroke Risk Screening and Education Program at a 400-bed regional medical center located in southeastern Alabama. The author outlines the implementation of a case management oriented program developed to target the specific high-risk populations residing in this rural area. Retrospective chart reviews were used to identify a large discrepancy between patient-recognized onset of stroke symptoms and the actual time it took for the patient to present to the Emergency Department (ED). A performance improvement goal was initiated to increase community awareness of stroke warning signs, stroke risk, and prevention. Summaries of the research process, design, and outcome data overwhelmingly indicated a positive correlation between stroke education and ED presentation time. PMID- 12777971 TI - Information management as a process and product of case management. AB - Information technology is an invaluable tool in case management practice management. Informed use of information technology can facilitate and enhance the work of case managers. This article reviews the usefulness of information management to nursing case management. The article discusses system requirement, security considerations, and database structure for a practice database. Voice recognition software, video conferencing, and use of the Web sites are discussed as additional practice enhancement. PMID- 12777972 TI - Providing relevant guidelines for case management practice: revised CMSA Standards of Practice for Case Management. PMID- 12777973 TI - Effect of a hypertension teaching protocol on patient and staff satisfaction in a rural community health center. AB - The purpose of this research was to examine the effect of a patient teaching protocol related to hypertension on patient and staff satisfaction in a rural community healthcare clinic. The rationale for this study was based on the positive relationship between staff and patient satisfaction highlighted in the literature. The literature also indicates that the implementation of a protocol can increase both patient and staff satisfaction. This study reinforces the positive relationship between patient and staff satisfaction in a rural setting. It also suggests a protocol may enhance patient and staff satisfaction. PMID- 12777974 TI - International case management in Japan: one case manager's story. AB - This article reviews one case manager's experience with international case management. It is a practical and useful view by an American case manager based in another country who developed and implemented a case management service for U.S. employees in Japan. The importance of cross-cultural relationships between hospitals, providers, and case managers is highlighted. Examples of cross cultural cooperation and international referrals are cited. PMID- 12777975 TI - The JAK/STAT pathway is not sufficient to sustain the antiproliferative response in an interferon-resistant human melanoma cell line. AB - The mechanism of resistance of malignant melanoma to treatment with interferon alpha is unknown, and currently there is no reliable method of predicting response. Signalling via the JAK/STAT pathway is known to mediate many interferon regulated events and has been implicated in mediating the antiproliferative response. The objective of this study was to determine whether defects in JAK/STAT signalling may be responsible for interferon resistance. The in vitro response to interferon was determined in a panel of established melanoma cell lines, and the components and functioning of the JAK/STAT pathway were examined in sensitive and resistant cell lines. Two melanoma cell lines, characterized as sensitive (MM418) and resistant (MeWo) to the antiproliferative effect of interferon, were both shown by Western blotting to possess all the protein components of the JAK/STAT pathway, and were shown to be capable of producing functional transcription factors using an electrophoretic mobility shift assay and a ribonuclease protection assay of known interferon-induced genes. In addition, both cell lines had intact antiviral and HLA upregulation responses. These data suggest that there is no defect in the JAK/STAT pathway per se in the MeWo cell line, and that the substantial resistance to interferon must be mediated through components either downstream or additional to this signalling pathway. Others have shown JAK/STAT defects to be responsible for interferon resistance in some melanoma cell lines. However, our results highlight the likely heterogeneity in the mechanisms leading to interferon resistance both in cell lines and tumours, and suggest that a clinical assay based on analysis of components of the JAK/STAT pathway may have only limited use as a predictor of interferon response. PMID- 12777976 TI - The effect of flavopiridol on the growth of p16+ and p16- melanoma cell lines. AB - Flavopiridol is the first cyclin-dependent kinase inhibitor to enter clinical trials. Flavopiridol has been shown to mimic, in part, the effect of the cell cycle control gene p16, which is frequently lost or mutated in malignant melanoma, making it an ideal candidate for targeted therapy in this disease. In these studies we investigated the effect of flavopiridol, at various concentrations, on the growth and gene expression of nine human melanoma cell lines with intact, absent or mutated p16. A cytostatic effect of flavopiridol on the growth of six melanoma cell lines with a mutated or non-expressed p16 (p16-) was seen at low concentrations of flavopiridol (mean 50% inhibitory concentration [IC(50)] = 12.5 nM), while the three melanoma cell lines with intact p16 (p16+) required higher concentrations (mean IC(50) = 25 nM) to produce this effect. Apoptotic cell death increased with increasing concentrations of flavopiridol in both p16- and p16+ cells. Exposure of cells to high flavopiridol concentrations (>100 nM) resulted in decreased expression of genes downstream in the normal p16 cell cycle control pathway (Rb and E2F) and the anti-apoptotic gene BCL2. No change in BCL2 expression was found after exposure to IC(50) concentrations of flavopiridol. These data indicate that flavopiridol in low, clinically achievable concentrations may have significant cytostatic effects, particularly in p16- melanoma cells, and may provide new molecular-based therapies for melanoma, particularly when combined with agents that target anti-apoptotic mechanisms. PMID- 12777977 TI - Autonomous histamine metabolism in human melanoma cells. AB - Melanoma cells constitutively produce various cytokines as well as growth factors and express their corresponding receptors. Exogenous histamine is known to be a growth factor for some tumours while in other cases histamine inhibits tumour growth, and acts on G protein-coupled H1 and H2 histamine receptors. In previous studies we have detected the expression of the l-histidine decarboxylase (HDC) gene and the presence of HDC protein in human melanoma cell lines. In the present study, the activities of the histamine-forming enzyme HDC and of the degrading enzymes diamine oxidase (DAO) and histamine N-methyltransferase (HNMT) were measured in primary (WM35 and WM983) and metastatic (M1 and HT168) human melanoma cell lines. HDC activity was found in WM35 and WM983 cell lines, while detectable HNMT activity was measured in WM983, M1 and HT168 lines. In contrast, DAO showed very low activity in melanoma cell lines. Melanoma cells release a detectable amount of histamine into the medium without external stimuli. These findings support the possibility of autonomous histamine metabolism in melanoma cells. Our results suggest that not only exogenous histamine but also histamine produced and released by the melanoma cells and acting as an autocrine and paracrine factor may influence cell proliferation and modulate the in situ immune response of the host. PMID- 12777978 TI - Establishment and characterization of human uveal malignant melanoma xenografts in nude mice. AB - The purpose of this study was to develop a suitable animal model for the investigation of the pathogenesis and therapy of uveal malignant melanoma. Eight choroidal malignant melanomas from eight patients were transplanted into nude mice in an attempt to establish a serially transplantable tumour model. Tumour tissue blocks (2 x 2 x 2 mm) from enucleated eyes with choroidal malignant melanoma were transplanted subcutaneously into the flanks of nude mice. The growing tumours were measured and serially transplanted. The tumour samples were investigated by histology, immunohistochemistry and electron microscopy. Only one of the eight transplanted primary tumours (13%) was established as a xenograft in nude mice. Furthermore, the take rate of the transplantable tumour was low (13%). The growth of the tumour fitted a Gompertz function, and the calculated tumour volume doubling time was 54 days. The transplanted tumour cells were epithelioid and slightly larger than the primary tumour cells and had prominent nucleoli. However, the transplanted tumour retained a morphological appearance similar to that of the primary tumour. Immunohistochemical examinations demonstrated that the cells preserved the characteristic properties of malignant melanoma. However, the transplanted cells demonstrated vimentin reactivity, whereas the primary tumour cells were negative for vimentin. It can be concluded that a new experimental model of malignant uveal melanoma with tumours that were easy to observe and access was established in nude mice. PMID- 12777980 TI - Rapid detection of metastatic melanoma in lymph nodes using proton magnetic resonance spectroscopy of fine needle aspiration biopsy specimens. AB - Accurate staging of patients with primary cutaneous melanoma includes assessment of regional lymph nodes for the presence of micrometastatic disease. Sentinel lymph node biopsy is highly accurate but is an invasive surgical procedure with a 5-10% complication rate, and requires labour-intensive and expensive histological examination to identify disease. A rapid, accurate and cost-effective non surgical technique able to detect micrometastatic deposits of melanoma in regional lymph nodes would be of great benefit. Fine needle aspiration biopsies and tissue specimens were obtained from lymph nodes from 18 patients undergoing node resection for metastatic melanoma and five patients undergoing radical retropubic prostatectomy. One-dimensional proton magnetic resonance spectroscopy was undertaken at 360 MHz (8.5 T). Lymph nodes were cut into 3 mm thick slices and embedded. Four sequential 5 microm tissue sections were cut from each block and stained, with haematoxylin and eosin, for S100 protein, for HMB45, and again with haematoxylin and eosin, respectively. Proton magnetic resonance spectroscopy distinguished between benign and malignant lymph node tissue (P < 0.001, separate t-test) and benign and malignant lymph node fine needle aspiration biopsy (P < 0.012) based on the ratio of the integrals of resonances from lipid/other metabolites (1.8-2.5 p.p.m. region) and 'choline' (3.1-3.3 p.p.m. region). In conclusion, one-dimensional proton magnetic resonance spectroscopy on a simple fine needle aspiration biopsy can distinguish lymph nodes containing metastatic melanoma from uninvolved nodes, providing a rapid, accurate and cost-effective non-surgical technique to assess regional lymph nodes in patients with melanoma. PMID- 12777979 TI - Association of TAP1 downregulation in human primary melanoma lesions with lack of spontaneous regression. AB - Spontaneous regression of primary melanoma lesions is regarded as the result of the recognition of melanoma-associated antigen (MAA)-derived peptides by cytotoxic T-lymphocytes and destruction of melanoma cells. The transporter associated with antigen processing (TAP1/2) is likely to play a crucial role in this process since it loads antigen peptides onto MHC class I molecules. To determine the impact of TAP defects on the spontaneous regression of melanoma lesions, we have compared the expression of TAP1 and TAP2 in 39 primary melanoma lesions exhibiting clinical and histological signs of tumour regression and in 35 primary melanoma lesions without regression phenomena. TAP1 expression was significantly associated with regression of melanoma lesions, since the staining pattern with anti-TAP1 antibody was positive in 38 of the 39 lesions exhibiting regression phenomena and in only 24 of the 35 lesions without histopathological signs of tumour regression. In the latter group, six lesions were stained with a heterogeneous pattern and five with a negative pattern. Furthermore, in lesions with a heterogeneous staining pattern, a clear association was found between TAP1 expression in melanoma cells and the presence of tumour-infiltrating lymphocytes. These results suggest that TAP1 plays an important role in the MAA-specific cytotoxic T-lymphocyte response, which has been suggested to underlie the spontaneous regression of primary melanoma. PMID- 12777981 TI - Dendritic cell density and activation status of tumour-infiltrating lymphocytes in metastatic human melanoma: possible implications for sentinel node metastases. AB - Nodal deposits of melanoma may present many years after resection of the primary tumour, implying initial suppression of tumour growth with subsequent immune escape. Using immunocytochemical techniques on frozen sections, the cellular types and activation status of infiltrating cells within a series of 19 clinically apparent nodal metastases of melanoma were studied. Infiltrating cells were assessed using a semiquantitative grading system. Macrophages (CD68+) and T lymphocytes (CD3+) (including both CD8+ and probably also CD4+ T-cells) were the predominant cells infiltrating the tumours. B-lymphocytes (CD20+) were generally present in low numbers. CD1a+ putative dendritic cell density and expression of the early lymphocyte activation markers interleukin-2 receptor alpha (IL2Ralpha) and CD69 was low. However, greater evidence of intermediate lymphocyte activation (CD38) was identified. Expression of interleukin-2 (IL2) by tumour-infiltrating cells was not detected. The paucity of staining for IL2 and IL2Ralpha, with greater expression of CD38 by infiltrating cells, suggests that the usual pathways of lymphocyte activation via IL2 were bypassed or impaired within the lymph node metastases. Low numbers of CD1a+ putative dendritic cells may result in reduced effector cell activation. These findings provide evidence to support the hypothesis that antitumour immune responses within clinically involved lymph nodes are reduced in metastatic melanoma. This also has possible implications for micrometastases to the sentinel lymph node. PMID- 12777982 TI - Proliferation rates in human malignant melanoma: relationship to clinicopathological features and outcome. AB - This prospective study investigated the clinical significance of cell kinetics, measured using bromodeoxyuridine injection and flow cytometry, in primary and metastatic cutaneous malignant melanoma. The findings illustrate that melanoma is a relatively slowly proliferating tumour, with a median potential doubling time (T(pot)) of 8.6 days. There were no significant differences in cell kinetics between primary and secondary disease. Both the duration of the S phase (T(s)) and T(pot), but not the labelling index (LI), showed a correlation with some of the main clinicopathological features in primary disease, including Breslow thickness. On univariate analysis, a short T(pot) and a high LI were associated with shorter disease-free intervals and overall survival for metastatic but not primary lesions. On multivariate analysis, these parameters retained significance when analysed separately with the main clinicopathological variables. These findings suggest that assessment of proliferation in melanoma is able to refine the available prognostic information and identify patients with a poor clinical outcome. PMID- 12777983 TI - Body site of cutaneous malignant melanoma--a study on patients with hereditary and multiple sporadic tumours. AB - Individuals with an increased risk of developing cutaneous malignant melanoma (CMM) include members of kindreds with hereditary cutaneous malignant melanoma (HCMM) and patients who have already been treated for a CMM. Some of these patients develop multiple primary cutaneous malignant melanomas (MCMMs). Ultraviolet radiation is the main instigator of CMM. There are indications that patients in these high-risk groups react differently to sunlight than patients who develop a single sporadic CMM. The objectives of this study were to analyse tumour site in patients with HCMM and sporadic MCMM. Data on 2517 patients with 2608 CMMs from a population-based regional cancer registry were used. The new computer program EssDoll was used for the analyses of primary tumour sites. This software is able to analyse any chosen body area(s) with reference to the number of tumours arising there. When the site of the first and second tumours in patients with sporadic MCMM were analysed in a skin 'field division', there was a significant concordance with respect to site (P < 0.0001). In patients with MCMM, the second primary tumour was significantly thinner than the first (P = 0.001). Primary tumour sites in patients with HCMM were compared with those in patients with a single sporadic CMM. In HCMM we found significantly fewer tumours in the head and neck area and more on the trunk. These differences remained significant in two different body area models, even when stratified for age (P < 0.05). In conclusion, a site-concordance was noted for sporadic MCMM. This may be the result of a 'field effect'. Our results indicate that intermittent ultraviolet exposure may be of relatively greater importance than chronic exposure in HCMM. PMID- 12777984 TI - Cyclooxygenase-2 promoter for tumour-specific targeting of adenoviral vectors to melanoma. AB - Novel therapeutic strategies are warranted for the treatment of metastatic melanoma as conventional therapies are inefficient. Conceptually, these strategies should be systemic and tumour-targeted. Gene therapy and viral oncolysis represent promising new approaches for cancer treatment that allow for the incorporation of molecular targeting strategies. In this regard, we analysed cyclooxygenase-2 (cox-2) expression as a potential new target for melanoma gene therapy. By reverse transcription polymerase chain reaction analysis, we showed cox-2 mRNA expression in all of the six tested melanoma cell lines, thus establishing cox-2 as a tumour marker for melanoma of potential interest for targeted therapeutics. Next, we analysed the activity and specificity of the cox 2 promoter within adenoviral vectors by luciferase assays. For this purpose, melanoma cell lines, primary melanoma cells and normal melanocytes were infected with adenoviruses containing cox-2 promoter sequences driving the luciferase reporter gene. The results demonstrated activity of the cox-2 promoter in melanoma cell lines and primary melanoma cells, but not in non-malignant primary epidermal melanocytes. Thus, we established herein the tumour specificity of the cox-2 promoter with potential applications for transcriptional targeting of adenoviral vector-based cancer gene therapy or virotherapy to melanoma. PMID- 12777985 TI - Phase I-II study on isolation antiblastic fotemustine perfusion after dacarbazine chemosensitization for advanced melanoma of the extremities. AB - Isolation limb perfusion (ILP) is the treatment of choice for locally advanced limb melanoma. With melphalan, the referral drug, complete response (CR) is achieved in about 50% of patients, but significant local toxicity occurs in up to 30%. The aim of the present phase I-II study was to challenge fotemustine (F) in ILP after systemic chemosensitization with dacarbazine (DTIC), given its lower toxicity and greater efficacy, as reported in a previous pilot study. Eleven patients with locally advanced limb melanoma were subdivided into triplets, and given F ILP at escalating doses (starting from 25 mg/l) after intravenous administration of 500 mg/m2 DTIC. Acute and chronic locoregional and systemic toxicity, tumour response and clinical outcome were evaluated. Two patients in the first triplet had G3-G4 local toxicity, so that the scheduled F dosage was halved. At drug levels of 12.5, 15.6 and 18.2 mg/l, local toxicity decreased, but only one of eight patients showed CR. The trial was then interrupted due to the low tolerability and poor efficacy of this perfusion regimen. At present, F ILP after DTIC chemosensitization should not be recommended for the treatment of locally advanced limb melanoma. PMID- 12777986 TI - Inefficacy of vindesine monotherapy in advanced stage IV malignant melanoma patients previously treated with other chemotherapeutic agents. AB - The anti-melanoma activity of vindesine as a single or polychemotherapeutic agent has been reported previously in adjuvant and first-line melanoma treatment. In this study, we investigated the usefulness of vindesine monotherapy as salvage therapy in stage IV melanoma patients after failure of other chemotherapies. Thirteen patients with progressive disease were treated with 3 mg/m2 vindesine every 2 weeks (median age, 61 years). Previous systemic treatment consisted of polychemotherapy or combined chemo-immunotherapy. All 13 patients suffered from visceral metastases (three lung, one liver, one adrenal gland and eight multiple visceral metastases). A median of three vindesine treatments was administered. Despite the various pre-treatments, the toxicity of vindesine was mild. In all 13 patients, vindesine treatment was stopped due to disease progression. The median survival after primary tumour diagnosis was 42 months (8-151 months), the survival after entering stage IV was 11 months (3-35 months), and the survival after starting vindesine therapy was 4 months (1-22 months). We conclude that vindesine monotherapy is ineffective in stage IV melanoma patients previously treated with other chemotherapeutic agents. PMID- 12777987 TI - Phase II evaluation of temozolomide in metastatic choroidal melanoma. AB - Temozolomide (Temodar) has demonstrated clinical activity against melanoma equivalent to that of intravenous dacarbazine (DTIC). Phase I clinical studies have shown that low dose chronic administration of temozolomide permits the delivery of higher dose intensities than a 5 day dose schedule. Temozolomide is hydrolysed to its active metabolite monomethyltriazenoimidazole carboxamide (MTIC) upon absorption from the gastrointestinal tract, while DTIC is inactive until it is metabolized in the liver to MTIC. In view of this, a higher concentration of MTIC will pass through the liver during the first pass when its source is temozolomide rather than DTIC. To determine if these characteristics of temozolomide will translate into a higher response rate than that achieved with DTIC, we conducted a phase II clinical trial of temozolomide in patients with uveal melanoma metastatic to the liver. Temozolomide was administered orally at a starting dose of 75 mg/m2 per day for 21 days every 4 weeks. Fourteen patients were enrolled in the trial. No complete or partial responses were observed. Stabilization of disease was achieved in two patients. The treatments were well tolerated. We conclude that, like DTIC, temozolomide at the dose and schedule studied in this trial is not effective for the control of metastatic melanoma of uveal origin. PMID- 12777988 TI - Treatment with histamine dihydrochloride and interleukin-2 in patients with advanced metastatic malignant melanoma: a detailed safety analysis. AB - Intravenous high dose bolus therapy with interleukin-2 (IL2) is associated with low overall response rates (15%) and significant toxicity. Phase II and III trials of a lower dose subcutaneous regimen of IL2 administered alone (n = 152) or in combination with histamine dihydrochloride (n = 239) have recently been completed. This article describes a comprehensive safety and toxicity analysis of the results of these two trials. The phase III trial demonstrated a survival benefit in patients with liver metastases in the histamine/IL2 arm. Eligible patients had stage IV malignant melanoma with at least one measurable lesion. Toxicity was graded using the National Cancer Institute (NCI) common toxicity scale. All reported adverse events were included in the analysis. Almost all toxicities in each treatment group were NCI grade 1 or 2. The incidence of toxicities expected to occur with histamine treatment, such as hypotension/vasodilation, headache and injection site reaction, were higher among patients receiving histamine. With the exception of headache, the incidence of grade 3 or 4 toxicities was similar across the treatment groups. The addition of histamine to the subcutaneous IL2 regimen did not result in a difference in the incidence of drug interruption, dose modification or discontinuation. Study related deaths were low and were not impacted by the addition of histamine to the IL2 regimen. PMID- 12777990 TI - Standard reference material for Her2 testing: report of a National Institute of Standards and Technology-sponsored Consensus Workshop. AB - A workshop was sponsored by the National Institute of Standards and Technology, the Cancer Diagnosis Program of the National Cancer Institute, the Food and Drug Administration, and the College of American Pathologists to address the need for a reference material for Her2 gene protein testing. It was agreed that such a standard was desirable and necessary to ensure the reliability of Her2 testing to qualify patients for trastuzumab therapy. Two standards consisting of well characterized cell lines will be produced, 1 that will be a National Institute of Standards and Technology-certifiable standard, and 1 that will be a commercially developed standard for use in all Her2 testing. It was also agreed that all Her2 testing must be performed on samples fixed only in 10% buffered formalin, as specified in the Food and Drug Administration-approved testing methods. Participants agreed to plan strategies to educate pathologists, clinicians, and laboratories about the need and use of such a standard. A National Committee for Clinical Laboratory Standards guideline for the use of the standard reference material will be created to facilitate this process. PMID- 12777989 TI - Melanoma patient staging: histopathological versus molecular evaluation of the sentinel node. AB - Lymphatic mapping and sentinel lymphadenectomy provide a minimally invasive means of directly determining the status of the regional lymph nodes in all patients who have a primary melanoma >1 mm thick but no clinical evidence of nodal involvement. Since the histological status of the sentinel node (SN) has been shown to be the most important prognostic factor in primary melanoma patients, the World Health Organization has recently recommended that sentinel lymphadenectomy should become the new standard of care for primary melanoma patients. This paper reviews the literature with regards to developments in and the current status of SN evaluation. Developments in the histopathological versus molecular detection of melanoma nodal metastases are reviewed, with specific emphasis on the strengths, limitations and clinical significance of these techniques. Molecular evaluation of the SN offers several advantages over standard histopathological analysis. These include an improved sensitivity, the cost-effective use of multiple markers for the improvement of detection rate and prognosis, as well as being less labour-intensive and costly. Moreover, molecular analysis has the potential to allow estimation of tumour burden. We review the potential causes of technical false-negative and false-positive reverse transcription-polymerase chain reaction (RT-PCR) results and how these could be eliminated by a systematic approach consisting of (i) careful and systematic assay design, which would include efficient tissue homogenization, choice of reagents and molecular markers, primer design and the use of one-stage versus two stage PCR; (ii) careful optimization of the RT-PCR parameters (in particular the PCR cycle number) through the use of appropriate control tissues; and (iii) aiming for high assay reproducibility and lastly by applying the necessary positive and negative controls with each batch of samples. We also review the significant improvement in patient prognosis and management that has been made possible by the development of sentinel lymphadenectomy and histopathological evaluation of the SN, and compare the clinical (predictive) value of histopathological analysis with that of RT-PCR. Although RT-PCR is able to detect additional, clinically significant SN metastases that are missed by routine histopathology, its current limitation is that it overestimates the number of patients who have clinically significant melanoma metastases. Therefore, we suggest and discuss appropriate steps that need to be taken in order to minimize these false-positives and make this molecular tool more acceptable for routine clinical use. PMID- 12777991 TI - Utility of immunohistochemistry in separating thymic neoplasms from germ cell tumors and metastatic lung cancer involving the anterior mediastinum. AB - A variety of primary and metastatic neoplasms can involve the anterior mediastinum and have similar radiographic or histologic appearances. We report an immunohistochemical (IHC) panel that includes CD5, placental-like alkaline phosphatase (PLAP), thyroid transcription factor 1 (TTF-1), cytokeratin (CK) 7, CK20, CK5/6, and CD57 for the separation of thymic neoplasms from germ cell tumors (GCTs) and metastatic lung malignancies. One hundred cases representing different types of neoplasms involving the anterior mediastinum were retrieved and consisted of 20 pulmonary adenocarcinomas (PAs), 20 pulmonary large cell carcinomas (PLCs), 20 pulmonary squamous cell carcinomas (PSCCs), 20 thymomas (Ths), 10 invasive thymomas (IThs), and 10 thymic carcinomas (TCs). Fifteen cases of GCTs were also included in the study. CD5 was positive in 17 of 20 PAs, 11 of 20 PLCs, 2 of 15 GCTs, and 9 of 10 TCs. PLAP was positive in 5 of 20 PAs, 2 of 20 PLCs, 1 of 20 PSCCs, and 11 of 15 GCTs. TTF-1 was positive in 18 of 20 PAs, 10 of 20 PLCs, and 2 of 20 PSCCs. CK7 was positive in 20 of 20 PAs, 15 of 20 PLCs, 8 of 20 PSCCs, 5 of 15 GCTs, 11 of 20 Ths, 7 of 10 IThs, and 4 of 10 TCs. CK20 was positive only in 2 of 20 PAs. CK5/6 was positive in all cases of PSCC, Th, ITh, and TC, in 9 of 20 PLCs, and 1 of 15 GCTs. CD57 was positive in 9 of 20 PAs, 3 of 20 PLCs, 8 of 15 GCTs, 13 of 20 Ths, and 6 of 10 IThs. In conclusion, CD5 had low specificity for TC (60%) because it also stained 85% of PA and 55% of PLC. CK5/6 was a sensitive but not a specific IHC marker for the epithelial component of thymic neoplasms (100%); TTF-1 had a good sensitivity (90%) and excellent specificity (100%) for distinguishing PA from thymic neoplasms. An IHC panel consisting of CD5, CK5/6, PLAP, TTF-1, and CD57 is useful for the separation of thymic neoplasms from other epithelial malignancies involving the mediastinum. PMID- 12777992 TI - Renal cell carcinoma antigen is expressed by yolk sac tumors and yolk sac elements of embryonal carcinomas. AB - Renal cell carcinoma antigen is a rather specific marker for normal and neoplastic renal tissue. We investigated the expression of this antigen in 34 gonadal and extragonadal germ cell tumors, including 8 pure yolk sac carcinomas and 26 embryonal carcinomas, 15 of which were combined with teratomas, seminomas, and dysgerminomas. Renal cell carcinoma antigen was demonstrated in all 8 yolk sac tumors and 21 of 26 embryonal carcinomas (81%). In yolk sac tumors, renal cell carcinoma antigen reactivity was diffusely present throughout the tumors. In embryonal carcinomas, this marker was identified only in yolk sac components. Both intracytoplasmic and membranous staining patterns were present. No reactivity was noticed in embryonal carcinoma cells, seminoma, dysgerminoma, and other components of teratomas. The study suggests an antigenic similarity between renal tubules and yolk sac tumors. Furthermore, the renal cell carcinoma antigen may be used as an addition to the panel of immunocytochemical markers for yolk sac carcinomas. PMID- 12777993 TI - Diagnostic utility of renal cell carcinoma marker in cytopathology. AB - Renal cell carcinoma (RCC) not uncommonly presents with metastases and causes diagnostic difficulty to the cytopathologist who is involved in the initial diagnostic workup of tumors with an unknown primary site. RCC marker (RCC Ma) recognizes a human proximal tubule antigen and was shown to have high specificity and relatively low sensitivity in preliminary studies on routinely processed tissue sections. We investigated the diagnostic usefulness of RCC Ma immunohistochemically in fine-needle aspiration (FNA) samples. A total of 34 FNA samples obtained from the following carcinomas were used: 7 RCCs, 5 metastatic RCCs, 4 hepatocellular carcinomas, 2 non-small cell carcinomas of the lung, 3 metastatic non-small cell carcinomas of the lung, 4 invasive ductal carcinomas of the breast, 2 pancreatic ductal adenocarcinomas, 4 metastatic transitional cell carcinomas of the urinary bladder, and 3 metastatic colon carcinomas. Routinely processed cell block sections of FNA specimens were stained with RCC Ma by using routine immunohistochemistry. Presence and distribution of staining were evaluated. Two of 7 (29%) primary and 2 of 5 (40%) metastatic RCCs showed immunoreactivity in less than 50% of carcinoma cells. Staining was focal, cytoplasmic, and granular. Scattered positive cells were present in two of the four hepatocellular carcinomas. All breast, lung, pancreas, colon, and transitional cell carcinomas were negative. RCC antibody has a low sensitivity (33%), most likely because of its focal staining pattern, and a high specificity (91%) in FNA specimens. Immunoreactivity in metastatic carcinoma of an unknown primary site, especially as part of a panel of antibodies, is useful in diagnostic cytopathology. RCC antibody has not been studied in hepatocellular carcinoma, and the significance of positivity observed in some of our cases is unclear. PMID- 12777994 TI - Assessment of Her-2/Neu status by immunohistochemistry and fluorescence in situ hybridization in mammary Paget disease and underlying carcinoma. AB - HER-2/Neu overexpression is seen in 20% to 30% of invasive breast carcinomas and has been reported in as many as 80% of high-grade infiltrating carcinomas. Earlier studies have suggested that 100% of the tumor cells in mammary Paget disease show overexpression of HER-2 protein. We undertook this study to assess HER-2 status of mammary Paget disease and of the underlying breast carcinoma, when present, by immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH). Formalin-fixed, paraffin-embedded tissue from 20 cases of mammary Paget disease were analyzed for HER-2 status by IHC and FISH. IHC for estrogen receptor (ER) was also performed. The patients ranged in age from 34 to 88 years, with a mean age of 62 years. Eighty percent of the cases showed strong overexpression (3+) of HER-2 protein by IHC, and all of these cases showed more than 5-fold amplification of the HER-2 gene by FISH. The remaining 4 cases, which were negative for HER-2/Neu by IHC, showed no amplification by FISH. All of the latter cases expressed ER, whereas no case that overexpressed HER-2 expressed ER. Sixteen cases had an underlying tumor, which was in situ in 6 cases. The underlying tumors were identical to the Paget disease with respect to their HER 2/Neu overexpression by both IHC and FISH. HER-2 overexpression was identified in 80% of our cases of Paget disease. There was 100% concordance between HER-2 protein overexpression by immunohistochemistry and gene amplification in both the Paget and the underlying tumor. Moreover, all of the cases negative for HER-2 overexpression expressed ER, whereas those positive for HER-2 did not. PMID- 12777995 TI - Zonula occludens-1 and Her-2/neu expression in invasive breast carcinoma. AB - A subset of breast carcinomas shows unexplained overexpression of HER-2/neu protein without HER-2/neu gene amplification. These tumors are often scored 2+ by immunohistochemistry (DAKO HercepTest). In vitro, Zonula occludens-1 (ZO-1), a tight junction protein, can upregulate HER-2/neu expression by sequestering a repressor of the HER-2/neu gene promoter. To assess this relation in vivo, we examined the expression of ZO-1 in 2 series of breast carcinomas. In group 1, 35 invasive breast carcinomas previously scored by HercepTest for HER-2/neu status (negative [n = 12], 2+ [n = 13], 3+ [n = 10]) were examined by immunohistochemistry for expression of ZO-1. ZO-1 expression was then correlated with HER-2/neu expression status. Twenty-four of 35 carcinomas (69%) were positive for ZO-1 expression. Frequency of ZO-1 expression did not differ between HER-2/neu-negative carcinomas (67%) and 2+ positive carcinomas (77%). Carcinomas with 3+ HER-2/neu expression were less commonly positive for ZO-1 (60%), but this difference was not significant. Twenty tumors contained ductal carcinoma in situ (DCIS); 18 of 20 DCIS (90%) were positive for ZO-1. Interestingly, in those tumors with ZO-1-positive DCIS, 4 of 18 (22%) were negative for ZO-1 expression in the invasive tumor. In group 2, 31 of 42 (74%) fluorescent in situ hybridization-nonamplified breast carcinomas showed ZO-1 expression, also with no significant difference in frequency between HER-2/neu-positive (by immunohistochemistry) and negative cases. ZO-1 expression does not correlate with HER-2/neu expression in breast carcinomas. Other causes of HER-2/neu protein overexpression should be sought in cases without HER-2/neu gene amplification. PMID- 12777996 TI - Lack of p53 nuclear immunostaining is not indicative of absence of TP53 gene mutations in colorectal adenocarcinomas. AB - Multiple studies using primary tumors have reported that alterations in p53 expression and detection of TP53 mutations are associated with clinical aggressiveness and poor response to specific therapies. However, there is no general agreement regarding the optimal technical approach to the analysis of p53. We have studied a series of 100 primary colorectal adenocarcinomas by immunohistochemistry with the monoclonal antibody PAb1801, and single-stranded conformation polymorphism (PCR-SSCP, exons 4-8) followed by direct sequencing of shifted bands. p53 Nuclear staining was undetectable (score 0) in 29 of 100 cases. However, gene mutations were detected in 15 of these cases, with all of these mutations leading to abnormal proteins. p53 Nuclear staining was detectable and scored as less than 10% tumor cells positive in 15 of 100 cases but was still considered to be displaying a p53-negative phenotype because the cut-off value for positivity was 10% positive tumor cells. Nevertheless, TP53 gene mutations were detected in 2 of these cases. p53 Nuclear immunoreactivities were detectable and scored as more than 10% tumor cells positive in 56 cases, considered the p53 positive phenotype. TP53 gene mutations were identified in 51 of these 56 cases. These results reveal that immunohistochemical assessment does not predict TP53 mutation status in colorectal adenocarcinoma, mainly in cases displaying absence of nuclear staining. It is thus concluded that molecular profiling should be conducted in parallel with immunophenotyping when analyzing colorectal tumors for p53 status. PMID- 12777997 TI - p27 and cyclin E/D2 associations in testicular germ cell tumors: implications for tumorigenesis. AB - The cyclins are key regulators of cell cycle progression and cellular proliferation. We have previously shown that in testicular germ cell tumors, cyclin E expression correlates with more aggressive tumors, higher clinical stage, and the presence of pulmonary metastases. Here, we have examined the association between cyclin activation and the proliferative rate of the pluripotential testicular tumor cell. We have shown that in a panel of 30 testicular germ cell tumors, 15 cases (50%) expressed the cyclin dependent kinase inhibitor p27; of note, 13 of 14 embryonal carcinomas (93%) coexpressed cyclin E and p27, suggesting inhibition of this cyclin. We show that 25 of 30 (83%) of the testicular germ cell tumors express cyclin D2. Using immunoprecipitation assays from the embryonal carcinoma cell line NTera2 or from tumor cell extracts, we have shown that cyclin D2 is complexed with p27, consistent with its known ability to sequester and block the cyclin E inhibitory function of p27. From these results, we propose a model in testicular germ cell tumors, in particular embryonal carcinomas, whereby the overexpression of cyclin D2, a gene localized on chromosome 12p--a region of DNA amplification in germ cell tumors--leads to the functional sequestration of p27 in the presence of cyclin E and cyclin D2, thus favoring cellular proliferation. PMID- 12777998 TI - Different localization of dendritic cell reservoirs in human immunodeficiency virus-1 subtype B versus subtype E-infected lymph nodes. AB - The presence of p24 protein was studied in lymph nodes from human immunodeficiency virus (HIV)-positive patients affected by persistent generalized lymphadenopathy. Paraffin-embedded lymph node sections from 50 HIV-1 subtype E infected lymph nodes from patients in Thailand and 25 HIV-1 presumably subtype B infected lymph nodes from patients in the United States were immunostained with p24 HIV major core and capsid monoclonal antibodies using the streptavidin-biotin immunoperoxidase technique. Positivity for HIV p24 protein was detected in 20 of 22 HIV-1 subtype B infected nodes in which lymphoid follicles were present, with p24 staining demonstrating a reticular pattern within the germinal centers. Interestingly, no case from 50 clade E-infected lymph nodes containing lymphoid follicles had such a reticular pattern in the germinal centers. This difference could be explained by differential infection of subsets of dendritic cells by the two HIV-1 clades, or perhaps by different routes of initial HIV-1 transmission. PMID- 12777999 TI - Gastric carcinoma with lymphoid stroma: incidence of EBV and Helicobacter pylori infection. AB - Lymphoepithelioma-like carcinoma (LELC) of the stomach is reported to be associated with Epstein-Barr virus (EBV) and to have a better prognosis than typical adenocarcinoma. However, the incidence of EBV and its association with Helicobacter pylori is controversial. In our series, 9 of 370 (2.4%) cases of gastric adenocarcinoma were LELC. Formalin-fixed, paraffin-embedded LELC tissue was immunostained with monoclonal antibodies against CD3, CD20, and Ki-67. Warthin-Starry stain for H. pylori and in situ hybridization for EBV were performed. As a control, 21 cases of adenocarcinoma with prominent lymphoid stroma were compared with LELC. The most common location of LELC was the body of the stomach (five cases), followed by the cardia (two cases), whereas the antrum was the most common site for the control group. There were no significant differences in age, sex, or lymph node metastasis between the LELC and control groups. The mean Ki-67 index was similar (38.2% in LELC versus 35.3% in control). H. pylori was found in 2 LELC cases (22%) and 6 controls (28.5%). EBV was positive in 7 LELC cases (77.8%) and 4 controls (19.0%). All patients with LELC are alive and well, whereas three patients have died of their disease in the control group. Although EBV appeared to be a contributing factor, H. pylori had a minor role in LELC. There were no clinicopathologic differences between LELC and the control group, except for the location of the tumor in the body of the stomach and the tendency toward a better prognosis in LELC. PMID- 12778000 TI - Immunohistochemical profile of serous papillary cystadenofibroma of the fallopian tube: a clue of paramesonephritic origin. AB - Cystadenofibromas of the fallopian tube are rare tumors of the female genital tract. These tumors are usually asymptomatic and are found incidentally. The authors present an incidentally found fallopian serous cystadenofibroma in a 48 year-old woman with leiomyoma uteri. The topographic localization of the lesion, histopathologic findings of mullerian-type epithelium, immunophenotypic profile of vimentin-cytokeratin coexpression, and diffuse apical epithelial membrane antigen (EMA) immunoreactivity suggested that the tumor was an embryologic remnant originating from the mullerian duct. PMID- 12778001 TI - Angiomyolipoma and PEComa are immunoreactive for MyoD1 in cell cytoplasmic staining pattern. AB - The family of tumors derived from mesenchymal perivascular epithelioid cells (so called PEComas) includes angiomyolipoma, lymphangioleiomyomatosis, clear cell sugar tumor of the lung, clear cell myomelanocytic tumor of ligamentum teres/falciform ligament, and abdominopelvic sarcoma of perivascular epithelioid cells. These tumors were characterized by coexpression of melanocytic (HMB-45) and muscle markers. MyoD1 transcription factor has crucial role in commitment and differentiation of mesenchymal progenitor cells to myogenic lineage. Antibodies to MyoD1 protein (nuclear immunoreactivity) have been shown highly valuable adjuncts in the diagnosis of rhabdomyosarcomas. To evaluate expression of the transcription factor MyoD1 in PEComas, we performed immunohistochemistry. Monoclonal antibody 5.8A for MyoD1 was used on a series of cases of formalin fixed, paraffin-embedded angiomyolipoma (n = 19), lymphangioleiomyomatosis (n = 3), clear cell sugar tumor of the lung (n = 1), and abdominopelvic sarcoma of perivascular epithelioid cells (n = 2). All cases showed strong granular immunostaining in the tumor cell cytoplasm with the anti-MyoD1 antibody. Cytoplasmic reactivity was noted in the spindle cells, fat cells, and epithelioid cells. Nuclei were negative in all tumors studied, and a clean background was obtained. Several normal and neoplastic human tissues have also been immunostained for MyoD1 without any positive cytoplasmic staining, with the exception of 2 alveolar soft part sarcomas. Cytoplasmic immunostaining with monoclonal antibody 5.8A for MyoD1 in PEComas may correspond to cross-reactivity with an undetermined cytoplasmic protein. Great caution should be exercised in interpreting the immunostaining results with anti-MyoD1 antibody 5.8A. PMID- 12778002 TI - Experimental brain injury induces activation of neural stem cells in the forebrain subependyma. AB - The subependymal zone (SEZ) of adult mammalians contains relatively quiescent neural stem cells that can be stimulated toward proliferation in response to specific stimuli. We used immunophenotypization to demarcate sharp boundaries of the SEZ and identify cell populations constituting the rat intact SEZ. Moreover, we studied the proliferation rates of SEZ cells under various experimental conditions that induced the lesion of the neighboring brain parenchyma or SEZ cells. Four groups of experimental animals included rats that were (1). mechanically injured, (2). intracerebrally injected with kainic acid, (3). treated with intracerebral injection of neurotoxic sodium nitroprusside, or (4). treated with intraperitoneal injection of cyclophosphamide. Animals were killed after 4 or 8 days. The number of SEZ proliferating cells was counted in coronal sections immunostained for proliferating cell nuclear antigen (PCNA) and bromodeoxyuridine. Our results show that all types of injury induced activation of SEZ neural stem cells, evidenced by the increase of corresponding proliferation indices when compared with intact brains. The increase was detected not only in ipsilateral but also in contralateral (intact) SEZ. After mechanically induced trauma of the right cerebral hemisphere, the increase in the number of SEZ proliferative cells was observed after 8 days in the right cerebral ventricle. Injection of kainic acid induced early responses in SEZ cells that reached the highest values. Injury induced by sodium nitroprusside evoked early increase of PCNA, whereas bromodeoxyuridine increase was detected in SEZ at day 8. Cyclophosphamide activated SEZ proliferation after 4 days, and the level of proliferation indices remained approximately the same at day 8. Our data suggest that each type of brain injury induces a SEZ proliferative response with a specific temporal pattern. PMID- 12778003 TI - Microarray as a model for quantitative visualization chemistry. AB - For visualization of proteins or nucleic acids, direct and indirect in situ fluorescence and absorption methods (immunohistochemistry and cytochemistry) have existed for many years. The authors describe a new experimental approach using microarray as a model to quantitatively compare both visualization methods. The spots obtained with the microarray robot had a progressive twofold decrease in concentrations and are used as objects with known amounts of DNA. Subsequent hybridization resulted in a direct fluorescence (DF) label or in hapten for indirect fluorescence (IF) and absorption. The results show that the image of the object in the IF method is larger than that in the DF method because of an edge effect, with stronger staining at the circumference. This leads to a higher plateau level and an 8- to 10-fold reduction in the detection threshold for IF compared with DF. These features are especially useful for one-color DNA-related microarray analysis, such as single nucleotide polymorphism, loss of heterozygosity, and mutation analysis, provided that the spots are not designed directly adjacent to each other, so that the edge effect is taken into account. The slope of the linear range for the IF method is much steeper than for the DF method, pointing to a narrow dynamic range in immunohistochemistry. It is noteworthy that the detection limit for absorption images after indirect immunoenzyme visualization is lower than for the DF images. The indirect immunohistochemistry semiquantitative absorption signal was at least similar compared with the DF fluorescence. In conclusion, an explanation for the difficulties experienced in quantitative immunohistochemistry is provided, and the data emphasize that in general, for daily pathology, semiquantitative patterns should suffice. Indirect labeling of DNA has useful characteristics for application in microarray analyses because of the large signal enhancement. PMID- 12778004 TI - Tissue macroarray: a simple and cost-effective method for high-throughput studies. AB - The tissue microarray technique is a powerful technique that can examine many clinical specimens in a single slide. However, this technique requires a specialized instrument for the construction of tissue microarray block. The authors describe an alternative tissue macroarray technique for the simultaneous analysis of multiple clinical specimens on a single slide. As an example of this method, the authors assembled 12 tumor samples from 12 different paraffin blocks on a glass slide and then immunostained with hMLH1 antibody. This simple method does not need specialized equipment and should be an alternative efficient approach for large-scale clinical specimens. PMID- 12778005 TI - HER-2 analysis in tissue microarrays of archival human breast cancer: comparison of immunohistochemistry and fluorescence in situ hybridization. AB - HER-2 gene alterations have been shown to have prognostic and predictive value for the treatment of breast cancer with therapeutic agents. As a result, the accurate evaluation of HER-2 status is crucial. HER-2 status is assessed at the protein level by immunohistochemistry and at the DNA level by fluorescence in situ hybridization (FISH). Although the best approach is to combine immunohistochemistry and FISH assays, doing so is not practical or cost-effective for routine histopathologic laboratories. The recent development of tissue microarray technology has allowed large-scale studies using formalin-fixed, paraffin-embedded material. We used this technique to assess HER-2 status in a cohort of 54 invasive breast cancer cases by immunohistochemistry and FISH assays to determine whether the results obtained were representative of the protein and gene expression patterns of the original whole tissue section. Concordance for HER-2 immunohistochemistry between the tissue microarray and full sections was 93%. Concordance for HER-2 FISH between the tissue microarray and full sections was 91%. Concordance between HER-2 FISH and HER-2 immunohistochemistry on the tissue microarray was 98%. We conclude that tissue microarrays provide highly comparable results in the assessment of HER-2 protein levels and allow large scale analysis of the HER-2 gene by FISH. PMID- 12778006 TI - Application of automated mRNA in situ hybridization for formalin-fixed, paraffin embedded mouse skin sections: effects of heat and enzyme pretreatment on mRNA signal detection. AB - Recently, an automated mRNA in situ hybridization application was introduced for the Ventana Discovery instrument. The application was designed so that all necessary steps from baking through signal detection were completed within 1 day on the instrument. We applied this technology for visualizing the expression site of versican in formalin-fixed mouse skin paraffin tissue sections. Our focus of this study was to demonstrate the effects of protease digestion or heating pretreatment, termed cell conditioning, on the hybridization signal using a well characterized versican antisense riboprobe. Paraffin sections were automatically deparaffinized, fixed, and acid-treated. Then, the tissue sections were subjected to protease digestion alone (3 strengths), cell conditioning alone, or the combination of cell conditioning and protease digestion. Hybridization was performed with digoxigenin-labeled versican antisense probe (20 ng/slide) for 6 hours, and the signal was detected using a Nitro blue Tetrazolium chloride 5 Bromo-4-cloro-3-indolyl phosphate toluidine salt (NBT/BCLIP) substrate solution for 3 hours on the instrument. Cell conditioning alone did not produce any signal, whereas the highest strength of protease digestion produced noticeable background staining. However, when cell conditioning and mild protease digestion were combined, the signal for versican mRNA was clearly demonstrated in the hair papilla region. Thus, we demonstrated the effects of the cell conditioning step followed by mild protease digestion for enhancing the mRNA target staining compared with protease digestion or the cell conditioning step alone. We verified that the automated in situ hybridization process was applicable for formalin fixed mouse skin paraffin sections and that the automated 1-day protocol is simple and reproducible. The precise control of automation allows fine tuning of temperature and enzyme dose to find the optimized assay condition for the signal to noise ratio and morphology. PMID- 12778007 TI - Differential protein analysis of spasomolytic polypeptide expressing metaplasia using laser capture microdissection and two-dimensional difference gel electrophoresis. AB - Full analysis of cellular protein constituents is a valuable tool in the evaluation of tissues. Traditional methods of evaluation, however, are time consuming and difficult to reproduce. Two-dimensional difference gel electrophoresis (2D-DIGE), a recently developed proteomic system, affords the ability to compare and evaluate protein extracts from multiple sources. Coupled with laser capture microdissection (LCM), this technology is a powerful tool in comparing the protein profiles of separate pure cell populations. Proteins are labeled in vitro with reactive cyanine dyes that fluoresce at differential wavelengths, and after comigration on two-dimensional gels, differing protein populations become apparent. The unique aspect of this technology is the ability to identify and quantify proteins from separate preparations without issues of gel-to-gel differences. These techniques coupled with the systems for robotic acquisition of specific spots on the gel, tryptic digestion, and MALDI mass spectrometry permit identification of proteins differentially expressed in two pure cell populations. The authors used these new technologies to analyze the protein constituents of spasmolytic polypeptide expressing metaplasia (SPEM), a gastric mucosal metaplasia (fundic antralization or pseudopyloric metaplasia) that develops in the atrophic fundus mucosa of mice infected with Helicobacter felis and in humans infected with Helicobacter pylori. In addition, SPEM has been identified in the atrophic mucosa surrounding a high percentage of gastric adenocarcinomas and may represent a precursor lineage of malignancy. This technology recognized 28 differentially expressed proteins between SPEM and surface cells. Identification of novel SPEM-related proteins would allow the development of new immunohistochemical antibodies to further study this important metaplasia. PMID- 12778008 TI - Effect of decalcification on the immunohistochemical expression of ABH blood group isoantigens. AB - Immunohistochemical stains for ABH blood group antigens were recently shown to be useful ancillary tools for sorting out specimen mix-ups in surgical pathology, irrespective of the fixatives used. However, the effects of decalcification on the expression of these antigens are not known. Therefore, to examine the validity of using ABH blood group immunohistochemistry in decalcified tissues, we studied the immunohistochemical expression of ABH blood group antigens in B5 fixed, decalcified, paraffin-embedded archival bone marrow specimens from 43 consecutive patients (13 blood group A, 6 group B, 20 group O, and 4 group AB). Immunohistochemical staining for A, B, and H blood group antigens was performed with monoclonal antibodies and an avidin-biotin detection method with citrate antigen retrieval. The results of immunohistochemistry were correlated with patients' blood groups as determined by serology. Immunohistochemical expression of the A and B blood group antigens with good staining intensity was detected in erythrocytes and endothelial cells. The immunoreactivity for H blood group antigen was attenuated by the decalcification process and could not be enhanced with citrate antigen retrieval. However, in all 43 cases, the A and B antigen staining pattern was concordant with patients' blood groups as determined with serology. These results show that decalcification does not have adverse effects on immunohistochemical expression of the A and B blood group antigens in tissue sections but may nullify the expression of H isoantigen. However, based on A and B immunoreactivity patterns, immunostaining for ABH blood group antigens can be helpful in resolving problems of specimen mix-ups and tissue floaters in decalcified specimens. PMID- 12778010 TI - The role of gamma knife radiosurgery in the treatment of primary and metastatic brain tumors. AB - With the widespread diffusion of stereotactic radiosurgical procedures, GKR treatments have gained considerable momentum as a major therapeutic option for patients harboring primary or metastatic brain tumors. Present results in high grade gliomas indicate a potential palliative role of this technique. The overall low radiosensitivity of these oncotypes and their infiltrative nature-with the resulting problems in properly defining the tumor target-are still a major obstacle to further development of the approach. In this regard, useful contributions are expected from advances in molecular neurobiology and functional neuroimaging as shown by preliminary investigations with MR spectroscopy. Surgery maintains a dominant role in the therapeutic armamentarium for low grade gliomas. However, in unfavorable cases (unresectable tumors, recurrences), GKR seems to be an effective alternative to conventional radiochemotherapy. In grade 2 astrocytomas and specifically in grade 1 pilocytic forms, short-to-mid-term reported studies have documented encouraging 70 to 93% local tumor control rates, with minimal cerebral toxicity. Finally, during the last decade, GKR has become a primary treatment choice for patients harboring small-to-medium-size brain metastases, with reasonable life expectancy and no impending intracranial hypertension. Focal tumor responses are consistently elevated, even in the most radioresistant oncotypes (melanoma, renal carcinoma); median and actuarial survival rates are far better than with conventional radiation treatments and are comparable to those observed in accurately selected surgical-radiation series. PMID- 12778011 TI - Recent advances in the molecular genetics of primary gliomas. AB - Primary brain tumors, particularly glioblastomas, remain a challenge for oncology. Uncontrolled cellular proliferation, lack of apoptosis, invasion, and angiogenesis are among the biologic processes that render these tumors both aggressive and difficult to treat. An understanding of the genetics and molecular events regulating these aggressive tumors is beginning to emerge, partly because of recent knowledge in genomics, gene expression analysis, and mouse tumor models. As a result, it is now generally accepted that brain tumors, particularly those arising from cells of glial lineage, result from stepwise accumulation of deleterious genetic alterations. Several genetic abnormalities have been described, and current research is aimed at elucidating their causal association with brain tumor formation and progression. The purpose of this review is to summarize some of the most important recently published findings on the molecular genetics of primary gliomas. PMID- 12778012 TI - Immunotherapy and biological modifiers for the treatment of malignant brain tumors. AB - The relative ineffectiveness of current therapies for malignant gliomas has led to the need for novel therapeutics. Therapies based on biologic modifiers are among a variety of cancer treatments currently in use or under experimental evaluation and have shown great promise, especially since several potent stimulators of the immune system have been cloned and are now available for clinical use. Early attempts at glioma therapy based on biologic modifiers, however, have failed to demonstrate significant effectiveness. In this review, we select and summarize the results of preclinical and clinical studies published during the past two years that focus on immunotherapy and biologic modifiers for treating gliomas. Despite limited clinical success, we conclude that an increased understanding of molecular biology and immunology from recent studies may pave the way for more effective approaches. PMID- 12778013 TI - Treatment of skull base tumors. AB - The rich multidisciplinary environment in which skull base surgery has developed has allowed for significant progress in the field and has improved patient outcomes. More precise and detailed anatomic dissection in the laboratories has provided surgeons with the benefit of undertaking the serious technical challenges encountered in the skull base. Significant work has been done and continues to be developed in the field of adjuvant treatments including radiotherapy, chemotherapy, and hormonal therapy. Using biologic end points and specific genetic targets in the tumors may further expand our management options in the future. PMID- 12778014 TI - Renal cell carcinoma. AB - Renal cell carcinoma (RCC) continues to present a diagnostic and therapeutic challenge. The increased use of abdominal imaging studies does not appear to completely account for the rising incidence of RCC. Alcohol consumption has been found to be a possible protective factor among women in a recent study, but among women with children, RCC risk may increase with each child born when compared with nulliparous women. An alternative staging system shows promise, and two randomized clinical trials clarify the role of removing the primary tumor in the setting of metastatic RCC. New agents have shown promise in early clinical trials such as CCI-779, pegylated interferon, thalidomide, and anti-VEGF antibody. PMID- 12778015 TI - Prostate cancer update. AB - Research has yielded important insights into the impact of radical prostatectomy in patients with localized prostate cancer. Other recent research has focused on the role of nutrition in prostate cancer development and progression, improved prognostication for patients with both early and advanced prostate cancer, efficacy of adjuvant and neoadjuvant hormonal therapy, and development of novel agents. In addition, the role of bisphosphonates in patients with bone metastasis was established. PMID- 12778016 TI - Pediatric genitourinary tumors. AB - Advances continue to be made in the evaluation and treatment of genitourinary tumors in children. Continued research on the cellular, molecular, and genetic process in tumorigenesis is rapidly evolving. We review the 2002 literature on pediatric genitourinary tumors. Molecular and cellular biology research continues to identify markers that may predict a worse prognosis. This identification will allow a more tailored approach with intensified treatment of aggressive tumors. PMID- 12778017 TI - Bladder cancer. AB - Bladder cancer is a significant public health problem responsible for more than 130,000 deaths annually worldwide. Disease prevalence is also remarkable, with more than 500,000 patients carrying the diagnosis in the United States alone. Significant progress has been made in understanding the underlying molecular and genetic events in bladder cancer. However, there remains a great need for the development of reliable markers that can provide clinically useful information regarding diagnosis and prognosis and to facilitate the selection of appropriate therapy in the individual patient. Ongoing and future investigation is anticipated to refine treatment of patients with high-risk superficial disease, to determine the role of neoadjuvant and adjuvant chemotherapy for high-risk invasive disease, and to improve the efficacy of chemotherapy for patients with metastatic bladder cancer. PMID- 12778018 TI - Germ cell tumors: review of selected studies from 2002. AB - Testicular cancer remains a major success story in the realm of solid tumors. There are still many remaining challenges in diagnosis, treatment, and prevention of long-term toxicity. Surgery and platinum-based chemotherapy, however, still encompass the main treatment modalities. Attempts to limit toxicity from both surgery and chemotherapy remain at the forefront of research. New chemotherapeutic options are available for patients with platinum-resistant disease, and stem cell transplant remains an area of active study. PMID- 12778019 TI - Sarcomas of the head and neck region. AB - PURPOSE OF REVIEW: This review discusses the classification, etiology, diagnosis, evaluation, treatment, and prognosis of sarcoma of the head and neck region. RECENT FINDINGS: Sarcomas account for less than 1% of all malignancies in the United States with only 5 to 15% of these sarcomas occurring in the head and neck region. However, about 1 in 3 pediatric sarcomas will occur in the head and neck region. Occasionally, these tumors are associated with genetic syndromes or previous radiation exposures, but, most commonly, no clear etiology exists. Pathologic classification is critical to the ultimate treatment and prognosis of sarcoma of the head and neck. Osteosarcoma, rhabdomyosarcoma, malignant fibrous histiocytoma, and angiosarcoma are the most common types of sarcoma to occur in the head and neck region; however, up to 20% of head and neck sarcomas will remain unclassified. Surgery has been central to the management of these malignancies with some exceptions in the pediatric population. Adjuvant chemotherapy is being utilized and/or studied for most high-grade sarcomas and adjuvant radiotherapy is important for disease control in high-grade soft-tissue sarcomas. Prognosis is clearly related to tumor grade and margin status. SUMMARY: Sarcomas of the head and neck region are rare malignancies often without a clear etiology. Expert pathologic review and classification is critical, as are quality imaging and multidisciplinary management. PMID- 12778020 TI - Simultaneous analysis of wall motion and coronary flow reserve of the left anterior descending coronary artery by transthoracic doppler echocardiography during dipyridamole stress echocardiography. AB - BACKGROUND: Coronary flow reserve (CFR) can be measured in the left anterior descending artery (LAD) by dipyridamole transthoracic Doppler echocardiography (DTTDE). This information may critically improve the diagnostic accuracy of dipyridamole stress echocardiography, which is limited by moderate sensitivity. OBJECTIVE: We sought to assess the feasibility and accuracy of value of DTTDE. METHODS: We evaluated 752 consecutive patients (478 men; mean age, 64.7 years) referred for dipyridamole stress echocardiography. The diastolic velocity in the LAD was continuously monitored with pulsed Doppler at baseline and during hyperemia induced by the infusion of 0.84 mg/kg of dipyridamole. CFR was calculated as the ratio of maximal and basal diastolic velocity; a value < 2 was considered to indicate decreased CFR. Wall motion was assessed semiquantitatively. Coronary angiography was performed in 132 patients the week after DTTDE. RESULTS: Adequate tracings were obtained in 95% of patients studied (715 of 752). In the subset of 132 patients undergoing coronary angiography, 71 patients (group A) showed a nonsignificant (<70%) and 61 patients (group B) a significant stenosis of the LAD. In group A, 65 had a normal wall-motion response (91.5% specificity) and 19 patients showed a decreased CFR (73.2% specificity; P =.1). In group B, new wall-motion abnormalities were seen in 42 patients (68.8% sensitivity), whereas CFR was decreased in 52 patients (86.8% sensitivity; P <.02). CONCLUSION: Simultaneous assessment of wall motion and CFR of the LAD with DTTDE was highly feasible and safe. The information about CFR had a significantly higher sensitivity than the analysis of wall motion during dipyridamole stress echocardiography. PMID- 12778021 TI - Which is the better method in detecting significant left anterior descending coronary artery stenosis during contrast-enhanced dobutamine stress echocardiography: coronary flow velocity reserve or wall-motion assessment? AB - The diagnostic accuracy of dobutamine stress echocardiography (DSE) depends on wall-motion assessment. Coronary flow velocity reserve (CFVR) during DSE can be measured by transthoracic Doppler echocardiography. To investigate comparative diagnostic accuracy between wall-motion and CFVR assessment in the detection of significant left anterior descending coronary artery (LAD) stenosis, 274 patients underwent both contrast-enhanced DSE and coronary angiography. Intravenous contrast agent, Levovist, was injected to enhance left ventricular endocardial border delineation and coronary flow velocity in the LAD. Wall motion was assessed by standard technique, and CFVR was calculated as coronary flow velocity at peak dobutamine stress divided by baseline coronary flow velocity by transthoracic Doppler echocardiography. CFVR could be successfully obtained in 232 patients (feasibility, 85%). After excluding 14 patients with isolated diagonal stenosis, CFVR was significantly lower in 65 patients with significant LAD stenosis than it was in 153 patients without stenosis (1.62 +/- 0.56 vs 2.72 +/- 0.94, P <.001). CFVR < or = 2.0 had a 75% sensitivity, a 81% specificity, and a 79% diagnostic accuracy for detecting significant LAD stenosis, and these values were comparable with those by wall-motion analysis (sensitivity, 78%; specificity, 89%; and diagnostic accuracy, 86%). The measurement of CFVR in the LAD during DSE was feasible and the diagnostic accuracy of CFVR was equivalent to wall-motion assessment in the detection of LAD stenosis. PMID- 12778022 TI - Mitral annulus motion compared with wall motion scoring index in the assessment of left ventricular ejection fraction. AB - The biplane disc summation method is the recommended echocardiographic procedure to determine left ventricular (LV) ejection fraction (EF). Assessment of mitral annulus motion (MAM) or wall motion scoring index (WMI) has been reported to be less dependent on image quality compared with the recommended method, and proposed as a surrogate to the disc summation method in calculation of LVEF. We aimed to compare MAM and WMI in the echocardiographic assessment of LVEF. In a randomly selected population-based sample of 75-year-old men and women in sinus rhythm (n = 409) MAM, as measured by M-mode, was compared with WMI, calculated as the mean value of wall motion scoring in 9 LV segments. LVEF, as measured by the biplane disc summation method was used as reference. The limits of agreement (mean difference +/- 1.96 SD) between LVEF and corresponding MAM values were -18 to +13 LVEF%, and between LVEF and corresponding WMI values were -12 to +13 LVEF%. The areas under the receiver operating characteristic curves for MAM and WMI to predict a LVEF < 50% were 0.892 and 0.998, respectively (95% confidence interval of the difference 0.062-0.149). The corresponding areas for MAM and WMI to predict a LVEF < 40% were 0.955 and 0.998, respectively (95% confidence interval of the difference 0.017-0.069). In conclusion, the ability of WMI to estimate LVEF was more favorable than MAM in this population-based sample of 75 year-old participants. The findings suggest that the WMI is preferable to MAM in estimating LVEF. PMID- 12778024 TI - Contrast echocardiography using intravenous octafluoropropane and real-time perfusion imaging predicts functional recovery after acute myocardial infarction. AB - Akinesia after acute myocardial infarction (MI) may be reversible, secondary to stunning, or irreversible, as a result of extensive myocyte necrosis. Distinguishing these 2 entities soon after MI is difficult, but has important clinical implications. The current study assessed the use of intravenous myocardial contrast echocardiography (MCE) in this setting. A total of 35 patients were studied 2 (+/- 1) days after an acute MI. Of these, 31 (91%) underwent myocardial revascularization. Perfusion was assessed using real-time MCE and an intravenous infusion of octafluoropropane microbubbles. Repeated echocardiograms were obtained 56 (+/- 29) days later. Normal perfusion predicted functional recovery with a positive predictive value of 66% and a negative predictive value of 81%. The accuracy of the technique was superior in myocardial segments supplied by the left anterior descending coronary artery (positive and negative predictive value: 70% and 90%, respectively). In multivariable analysis, the mean MCE perfusion score in akinetic segments was the most powerful independent predictor of functional recovery (odds ratio 8.6, P =.02). These data suggest that real-time intravenous MCE is a useful predictor of functional recovery of akinetic myocardium after acute MI. PMID- 12778023 TI - Early changes in left ventricular volume and function are predictors for long term remodeling in patients with acute transmural myocardial infarction and preserved systolic function. AB - BACKGROUND: We sought to describe the degree of long-term left ventricular (LV) remodeling after acute transmural myocardial infarction with preserved LV systolic function, and to evaluate whether Doppler echocardiographic parameters in the early phase could predict this process. METHODS: A total of 60 patients without heart failure and with LV ejection fraction > or = 0.40 (mean 0.48 +/- 0.054), were followed up with Doppler echocardiographic examinations at baseline, 3 months, and 1 and 2 years. RESULTS: There was a significant increase in LV end diastolic volume index of 7% (P =.006) and LV end-systolic volume index of 8% (P =.03), and no change in ejection fraction. This remodeling was confined to 7 patients (12%) with a significant increase in LV end-diastolic volume index above 20 mL/m(2). There was also a significant increase in the deceleration time of both the early mitral filling wave (Delta early mitral filling wave = 58 milliseconds, P <.0005) and the diastolic forward component of pulmonary venous flow (Delta diastolic forward component of pulmonary venous flow = 61 milliseconds, P <.0005), and a shift in filling pattern with increasing prevalence of abnormal relaxation. Changes in end-diastolic volume index were predicted by baseline early mitral filling wave less than 100 milliseconds, but the most powerful predictors of 2-year remodeling were volume changes at 3 months. CONCLUSION: Twelve percent of patients with Q-wave infarction and ejection fraction > or = 0.40 experienced significant LV dilatation at 2 years, and this late remodeling was partly related to baseline filling characteristics. PMID- 12778025 TI - Force-frequency relationship in the echocardiography laboratory: a noninvasive assessment of Bowditch treppe? AB - BACKGROUND: Estimation of contractility of the left ventricle is an important, and as yet elusive, goal with noninvasive techniques. OBJECTIVE: We sought to assess the feasibility of a totally noninvasive estimation of force-frequency relation (FFR) during exercise stress in the echocardiography laboratory. METHODS: We enrolled 13 healthy control patients (12 men, age 38 +/- 15 years) as group I, and 50 patients (38 men, age 64 +/- 11 years) referred for exercise echocardiography as group II. To build the FFR, the force was determined at each step as the ratio of the systolic pressure (cuff sphygmomanometer)/end-systolic volume index (biplane Simpson's rule/body surface area). The slope of the relationship was calculated with the linear best fit of the FFR. RESULTS: Noninvasive systolic pressure/end-systolic volume ratio was obtained in all patients. The slope of the linear best fit of the force-frequency curve was lower in group II compared with group I (group II = 10.1 +/- 9.3 x 10(-2) vs group I = 14.9 +/- 9.9 x 10(-2) group I, P =.04). By regional wall-motion analysis, 2 subgroups were identified in group II: group IIA (n = 8) had a positive echocardiogram; and group IIB (n = 42) had a negative echocardiogram. The slope of the force-frequency curve was lower in patients with ischemia compared with those without (group IIA = 3.5 +/- 4.2 x 10(-2) vs group IIB = 11.4 +/- 9.5 x 10( 2); P =.012). Heart rate-systolic pressure/end-systolic volume index relation was biphasic, with an initial positive slope and a subsequent negative slope in 1 patient of group I, 4 patients of group IIA, and 15 patients of group IIB (P <.05 vs group I). CONCLUSION: A noninvasive estimation of FFR can be easily determined during exercise echocardiography. This index of global contractility is theoretically appealing for identification of limited contractile reserve and latent global left ventricular dysfunction. PMID- 12778026 TI - Acute reductions in ventricular myocardial tissue velocities after direct current cardioversion of atrial fibrillation. AB - BACKGROUND: Cardioversion by direct current (DC) and other methods can cause atrial "stunning." There are case reports of acute pulmonary edema after DC cardioversion, but whether acute ventricular dysfunction is a general consequence of DC cardioversion is unknown. We have investigated whether DC cardioversion acutely affects myocardial velocity assessed by Doppler tissue imaging. METHODS: 40 patients (30 with atrial fibrillation and 10 with atrial flutter) undergoing elective DC cardioversion underwent transthoracic echocardiography with Doppler tissue imaging before and immediately after cardioversion, and after follow-up. Peak systolic velocity was derived for 6 ventricular segments using Doppler tissue imaging. RESULTS: Immediately after DC cardioversion of atrial fibrillation, peak systolic velocity decreased in basal lateral (4.3 +/- 2.0-3.3 +/- 1.7 cm/s, P <.001), mitral annulus-septal (3.8 +/- 1.0-3.5 +/- 0.9, P <.05), mitral annulus-lateral (4.9 +/- 1.6-4.1 +/- 1.7, P <.001), and tricuspid annular (7.8 +/- 2.0-7.0 +/- 1.2, P <.03) segments, even though left ventricular ejection fraction was unchanged. In contrast, for the atrial flutter group there were no significant changes in peak systolic velocity in any segment post-DC cardioversion. Follow up studies were performed after sustained in sinus rhythm in both atrial fibrillation and atrial flutter groups. For both groups, increased peak systolic velocity was found in all 6 segments on follow-up (all P <.05). CONCLUSIONS: DC cardioversion causes subclinical, acute reversible reduction in left ventricular peak systolic velocity in patients with atrial fibrillation. The causes of this reduction in myocardial contractile velocity and the circumstances in which acute dysfunction become clinically significant warrant further investigation. PMID- 12778027 TI - Left ventricular filling patterns and pulmonary artery pressures in patients aged 90 to 100 years with normal echocardiography results. AB - Left ventricular filling and pulmonary artery pressure (PAP) have not been well described in patients > or = 90 years old. We reviewed our database to obtain echocardiograms with normal findings in this age group. The peak early and late mitral filling velocities, the early and late velocity-time integral (VTI), the mitral deceleration time, and the atrial filling fraction (late VTI/total VTI) were measured. PAP was obtained from the peak tricuspid regurgitation velocity. Of 252 echocardiograms performed on patients > or = 90 years old, 47 (19%) were normal. The mean +/- SD values were: ratio of peak early and late mitral filling velocities, 0.69 +/- 0.16; deceleration time, 240 milliseconds +/- 31; ratio of early and late VTI, 1.0 +/- 0.3; atrial filling fraction, 47% +/- 7; and PAP, 31 mm Hg +/- 6. Left ventricular filling in patients aged 90 to 100 years demonstrates increasingly impaired left ventricular relaxation from patients in their eighth and ninth decades. PAP continues to increase with age and PAP < or = 40 mm Hg in the elderly could be considered normal. PMID- 12778028 TI - Left ventricular systolic torsion and exercise in normal hearts. AB - This study examines the use of a novel 2-dimensional echocardiographic technique to measure left ventricular (LV) systolic torsion or twist in normal human hearts during maximal exercise. The effects of dynamic exercise on LV twist have not previously been determined. LV twist was measured in 25 healthy human control patients before and after maximal treadmill exercise. LV end-systolic volume decreased and LV ejection fraction increased in a predictable manner. Systolic twist increased by 8.4 +/- 2.8 degrees (86%). Twist is believed to store potential energy and to play an important role in generating diastolic suction. The increase in twist observed may play an important role in facilitating LV diastolic filling during maximal exercise. PMID- 12778029 TI - Three-dimensional visual guidance improves the accuracy of calculating right ventricular volume with two-dimensional echocardiography. AB - Three-dimensional guidance programs have been shown to increase the reproducibility of 2-dimensional (2D) left ventricular volume calculations, but these systems have not been tested in 2D measurements of the right ventricle. Using magnetic fields to identify the probe location, we developed a new 3 dimensional guidance system that displays the line of intersection, the plane of intersection, and the numeric angle of intersection between the current image plane and previously saved scout views. When used by both an experienced and an inexperienced sonographer, this guidance system increases the accuracy of the 2D right ventricular volume measurements using a monoplane pyramidal model. Furthermore, a reconstruction of the right ventricle, with a computed volume similar to the calculated 2D volume, can be displayed quickly by tracing a few anatomic structures on 2D scans. PMID- 12778030 TI - Feasibility of transesophageal echocardiography with a ten-French monoplane probe. AB - OBJECTIVES: We examined the feasibility of transesophageal echocardiography (TEE) using a 10F monoplane probe developed for intracardiac ultrasound (AcuNav, Acuson/Siemens, Mountain View, Calif). BACKGROUND: Traditional TEE uses a 10- to 12-mm-diameter probe, and conscious sedation is customary to minimize patient discomfort. Because of its small size (3.2-mm diameter), the 10F monoplane probe can be inserted into the esophagus using only topical anesthesia. This provides the potential for a more easily tolerated examination. METHODS: A total of 20 patients underwent a comprehensive TEE using an adult multiplane probe. Immediately afterward, the 10F monoplane probe was inserted into the esophagus and a targeted examination completed. The 10F monoplane studies were blindly reviewed by 3 observers for the study indication and for 16 diagnostic elements. These were graded against an expert's review of standard TEE. RESULTS: The 10F monoplane probe was well tolerated in all patients. Observers A, B, and C answered the clinical question in 80%, 85%, and 100%, respectively, with the 10F probe. The percentage of clinical elements deemed evaluable was 71%, 78%, and 80%, respectively. Limitations included incomplete visualization of the mitral valve and a systematic underestimation of the severity of valve regurgitation. CONCLUSIONS: The 10F monoplane probe is safe, well-tolerated, and capable of evaluating many clinical questions. Because of its small size, conscious sedation may not be necessary. It may be useful for targeted evaluations, for monitoring invasive procedures, or for intermediate or long-term monitoring in an intensive care department. PMID- 12778031 TI - Transesophageal echocardiography monitoring in the intensive care department: the management of hemodynamic instability secondary to thoracic tamponade after single lung transplantation. AB - A 65-year-old woman with pulmonary fibrosis had hemodynamic instability develop after single lung transplantation. Transesophageal echocardiography was useful in determining the cause of hemodynamic instability, which was secondary to thoracic tamponade. Furthermore, transesophageal echocardiography monitoring was important in the decision to initiate extracorporeal membrane oxygenation to evaluate adequate positioning of the cannula and in the extracorporeal membrane oxygenation weaning process. PMID- 12778032 TI - Unusual right ventricular flow pattern in a patient with inferior wall myocardial infarction. AB - We describe a case of an unusual right ventricular flow pattern in a patient with inferior wall myocardial infarction associated with massive right ventricular infarction. Echocardiography was significant for nearly continuous antegrade flow through both the tricuspid and pulmonic valves demonstrated by color and pulsed wave Doppler. The tricuspid and pulmonic valves remained open throughout the cardiac cycle. These findings signify the nearly complete loss of right ventricular pumping function, and indicate that the right heart was serving as a passive conduit with a flow pattern similar to a vein. PMID- 12778033 TI - Both diastolic and systolic function alternate in pulsus alternans: a case report and review. AB - Pulsus alternans is occasionally seen in the setting of heart failure. This case describes a patient with both these findings, and alternating diastolic and systolic left ventricular function on color Doppler and Doppler tissue imaging. PMID- 12778034 TI - Diastolic ventricular dysfunction as a marker for hypertrophic cardiomyopathy in a family with a novel alpha-tropomyosin mutation. AB - BACKGROUND: Early identification of familial cases of hypertrophic cardiomyopathy (HCM) depends on screening echocardiography, but hypertrophy may not be the most sensitive marker for the disease. We report the echocardiographic findings of a family with HCM and a newly reported mutation in the gene (TPM1) encoding alpha tropomyosin.Methods and results An 8-year-old girl had sudden cardiac death, and was found to have HCM and a novel L185R-TPM1 mutation on postmortem examination. Screening echocardiograms and DNA analyses were performed on her family. Of the 5 remaining family members, 3 were genetically affected. Those without the TPM1 mutation had normal echocardiographic results. The only echocardiographic finding that identified all 3 of the gene-positive family members was an abnormal left ventricular diastolic filling pattern. CONCLUSION: Abnormal left ventricular diastolic filling patterns, indicating diastolic dysfunction, may provide an early marker for the diagnosis of familial HCM in children, even in the absence of left ventricular hypertrophy. PMID- 12778036 TI - Aging issues moving mainstream. PMID- 12778038 TI - Food biosecurity. PMID- 12778039 TI - Predictors and consequences of food neophobia and pickiness in young girls. AB - OBJECTIVE: Vegetable intake among children is well below recommended levels. We assessed whether food neophobia and pickiness contribute to low vegetable intake in school-aged girls and if there are distinct predictors for neophobia and pickiness. Children with food neophobia are reluctant to eat new foods whereas picky children resist eating many familiar foods. Design/subjects Participants were 192 7-year-old girls and their parents, recruited for a study of girls' nutrition and development. We examined relationships between food neophobia and pickiness and assessed whether these variables predicted girls' vegetable consumption and predictors of food neophobia and pickiness. ANALYSES: The data were analyzed using a two-step process. First, we used a two-way analysis of variance to assess whether girls who scored high or low on food neophobia and pickiness measures had different levels of vegetable consumption. We used multiple regression analysis to determine predictors of food neophobia and pickiness in the girls. RESULTS: Girls with both food neophobia and pickiness consumed fewer vegetables (1.1+/-0.1) than girls with neither neophobia nor pickiness (1.6+/-0.1). Neophobia and pickiness were modestly related in this sample, but had different predictors. Girls with food neophobia were more anxious and had mothers with food neophobia. Picky girls had mothers with less variety in their vegetable intake (r=-0.22) and mothers who perceived their family to have little time to eat healthful foods (r=0.36). In addition, picky eaters were breastfed for fewer than 6 months (r=-0.25). Pickiness was predicted primarily by environmental or experiential factors subject to changes; neophobia was predicted by more enduring and dispositional factors. APPLICATIONS: Because food neophobia and pickiness negatively influence vegetable intake, intervention strategies to increase vegetable intake should focus on predictors of neophobia and pickiness, especially those subject to change. PMID- 12778040 TI - Baseline fat-related dietary behaviors of white, Hispanic, and black participants in a cholesterol screening and education project in New England. AB - OBJECTIVE: To examine baseline fat-related dietary behaviors of white, Hispanic, and black participants in Minimal Contact Education for Cholesterol Change, a National Institutes for Health-funded cholesterol screening and education project conducted in New England. SUBJECTS: A sample of 9,803 participants who joined the study at baseline (n=7,817 white; n=1,425 Hispanic; and n=561 black). METHODS: Participants completed baseline questionnaires that included demographic and psychosocial items as well as the Food Habits Questionnaire, a dietary assessment tool measuring fat-related dietary behaviors. They also had their blood cholesterol level and height and weight measured. STATISTICAL ANALYSES: Analysis of variance (ANOVA) was used to compare racial/ethnic groups on continuous demographic variables, and the chi(2) test of association was used to compare groups on demographic categorical variables. Multivariate analysis of variance (MANOVA) was used to compare mean differences between racial/ethnic groups on six behavioral subscales (Fat Factors) differentiating domains of behavior related to fat intake and to compare 27 individual fat-related dietary behaviors. RESULTS: After adjusting for sex, age, marital status, education, employment status, and percent time lived in the United States, white participants had the lowest Food Habits Questionnaire summary score (2.44) (indicating a lower fat diet), followed by Hispanic (2.61) and black (2.68) participants. The three ethnic groups also differed on the prevalence of Fat Factors and specific fat-related dietary behaviors. White participants were more likely to use lower-fat alternatives, to avoid frying, to replace meat, and to modify meat to make it lower in fat. However, they were least likely to eat fruits and vegetables for snacks and desserts. Hispanic participants were more likely to engage in fat-avoidance behaviors and to eat fruits and vegetables for snacks and desserts. Black participants were less likely to eat meatless meals and modify meats to make them lower in fat. Black and Hispanic participants were more likely than white participants to fry foods. Hispanics were less likely to read food labels for nutrition information. The most and least prevalent fat-related behaviors also differed by each ethnic group, showing that different behaviors were more and less easily implemented by each ethnic group. CONCLUSIONS/APPLICATIONS: The results of this study suggest that there is a need for improvement in dietary behaviors related to fat intake, especially for blacks and Hispanics, and that the specific dietary behavior issues differ widely by ethnicity. These results can be used by nutrition educators and researchers to help them decide what messages to emphasize in dietary counseling, nutrition education programs, and materials. The results can also be used to help design better dietary assessment tools and more effective interventions for culturally diverse populations. PMID- 12778042 TI - What are some resources that can help my clients sort through the conflicting information on dietary supplements? PMID- 12778041 TI - A prospective study: growth and nutritional status of children treated with the ketogenic diet. AB - OBJECTIVE: To assess the nutritional status of children treated with the classic and medium-chain triglyceride (MCT) ketogenic diets. DESIGN: A prospective, nonrandomized study design was used to measure nutrient intakes, growth, and biochemical indexes of children, age 1 to 16 years, with intractable epilepsy before and after 4 months' treatment with the classic and MCT ketogenic diets. None of the children had been on earlier dietary regimens. SUBJECTS: Of 58 children asked to participate in the study between September 1998 and July 2000, consent was obtained for 30 children. Fourteen children on the classic diet and 11 children on the MCT diet completed the study (83% completed). Statistical analysis performed Paired t tests were done on anthropometric and biochemical indexes. Nutrient intakes were compared with Dietary Reference Intakes (DRIs). RESULTS: Both groups had statistically significant height increases of 2 to 3 cm (P<.05), but did not have significant increases in height/age percentiles. Weight percentiles decreased by approximately 10 percentiles for both diets; P=.043 for classic diet and.051 for MCT diet. Nutrient intakes from the diet and vitamin and mineral supplements met the DRIs except for phosphorus (both diets) and folate (classic diet). All biochemical indexes, including albumin, remained within the normal range. For the MCT diet, there was a 0.7 decrease in the ratio of total cholesterol to high-density lipoprotein ratios (P<.0009) at 4 months. APPLICATIONS: When treating children on a ketogenic diet, clinicians should recommend adequate intake of energy and protein, a higher proportion of unsaturated to saturated dietary fats, and consider vitamin and mineral supplements. PMID- 12778043 TI - Relative validation of a beverage frequency questionnaire in children ages 6 months through 5 years using 3-day food and beverage diaries. AB - OBJECTIVE: To determine the relative validity of a quantitative beverage frequency questionnaire in assessing beverage, calcium, and vitamin D intakes using 3-day food diaries for reference. DESIGN: Parents were asked to complete questionnaires for the preceding week and diaries for the following week for their children. Calcium and vitamin D intakes were estimated from human milk, infant formulas, and cow's milk ("beverages") for questionnaires and diaries and from "all foods and beverages" for diaries. Data collected at 6 and 12 months and 3 and 5 years of age as part of the Iowa Fluoride Study (N=700) were analyzed cross-sectionally. SUBJECTS: Children (N=240); 60 randomly selected from each quartile of energy intake at 6 months of age. STATISTICAL ANALYSES: Spearman correlation coefficients, weighted kappa statistics, and percentages of exact agreement were used to assess associations between tools. RESULTS: Correlations between mean daily beverage intakes estimated from questionnaires and diaries ranged from 0.95-0.99 for human milk, 0.84-0.85 for infant formula, 0.63-0.86 for cow's milk, 0.54-0.69 for juice/drinks, 0.26-0.59 for liquid soft drinks, 0.35 0.74 for powdered soft drinks and 0.54-0.70 for water. Correlations between mean daily nutrient intakes estimated from questionnaires and diaries "beverages" ranged from 0.64-0.74 for calcium and 0.60-0.80 for vitamin D; and between questionnaires and diaries "all foods and beverages" ranged from 0.41-0.63 for calcium and 0.43-0.80 for vitamin D. APPLICATIONS: A quantitative beverage frequency questionnaire can provide a relative estimate of beverage, calcium, and vitamin D intakes. PMID- 12778044 TI - Developing health messages: qualitative studies with children, parents, and teachers help identify communications opportunities for healthful lifestyles and the prevention of obesity. AB - Childhood overweight is at an all-time high in the United States. In an effort to better understand children's, parents', and teachers' attitudes, perceptions, and behaviors about preventing overweight in childhood and to explore potential avenues for communicating overweight prevention messages, we conducted qualitative research with these three groups in 2000. Our research consisted of three progressive phases, each building on information obtained from the previous phase: Phase 1, 16 focus groups (N = 112); Phase 2, in-home observations, in depth interviews, and diaries (N = 6 families); Phase 3, 10 qualitative interview sessions (N = 46). Both parents and children indicated that encouragement and "small victories" to sustain involvement in getting more fit were critical to success. The findings also suggest that children need direct messages to motivate them to change their exercise and eating habits, as well as tips on cooperating with their parents to achieve fitness goals. Parents need to learn how to talk about eating and exercise habits with their children in positive and encouraging ways and to learn how to help their children maintain efforts to get fit. Teachers consider it essential that parents support healthful lifestyles at home. Parents and children need positive, realistic approaches to getting fit, such as answers to questions about healthful lifestyles; ideas for physical games and activities the family can enjoy together; attainable goals and small steps to healthful eating; healthful meal, snack, and recipe suggestions; incentive ideas for getting kids active; and referral services for local support groups. Parents and children need to work together in addressing the overweight prevention issue and need effective tools to facilitate this cooperative effort. PMID- 12778045 TI - An overview of methodologies, proficiencies, and training resources for controlled feeding studies. AB - Dietary intervention studies of human beings produce valuable information regarding dietary effects on biological processes and risk factors for chronic diseases. Using the well-controlled feeding approach, participants consume only foods that have been precisely prepared in a research kitchen, whereas in behavioral counseling studies, participants self-select their foods within guidelines. Because controlled feeding studies meticulously control experimental diets, they are intellectually and logistically challenging to conduct. They afford exciting opportunities for dietetic professionals in designing protocols, developing budgets, and collaborating in multidisciplinary research teams. Research dietitians use food composition data and chemical analysis of menus to prepare research diets with precision. They determine the energy requirements of subjects and adjust diets as required, most often for weight maintenance, throughout the study. All people involved in research must be attentive to the ethical treatment of the study participants while motivating them to adhere to the protocol requirements. Dietitians possess many of these skills, but may require training specific to well-controlled feeding studies. Information related to the conduct of controlled feeding studies has recently become more accessible. We provide an overview of well-controlled feeding study methodologies, proficiencies for planning and implementing these studies, and training resources. PMID- 12778046 TI - History and evolution of a successful certification program in nutrition support: the CNSD experience. AB - As early as the 1970s, articles were published on the role of dietitians in nutrition support. Both the American Society for Parenteral and Enteral Nutrition and The American Dietetic Association addressed the issues of specialization and certification. Development of a specialty area credential requires a strong demand by practicing clinicians, a unique area of clinical practice with a distinct body of knowledge and the commitment to evolve with clinical practice. Numerous dietitians were involved in the inception, development, and leadership that brought forth certification in nutrition support. This article documents the efforts and participation of a dedicated group of professionals whose common goals resulted in the establishment of a successful, sustaining certification in a specialty area of clinical dietetics: certified nutrition support dietitian. The certified nutrition support dietitian program is now in its 15th year with 2,000 dietitians currently certified both nationally and internationally. A primary focus of the credential is to recognize minimum competency for dietitians practicing in the field of enteral and parenteral nutrition and to provide safe and effective nutrition support therapy. This article provides a framework for persons in other specialty areas attempting to develop certification programs. PMID- 12778047 TI - Lower bone mass detected at femoral neck and lumbar spine in lower-weight vs normal-weight small-boned women. AB - Sixty-one nonsmoking, healthy, young, menstruating women aged 18 to 30 years generally considered at peak skeletal bone mass were screened for diseases and drugs known to adversely affect bone mineral density (BMD). Anthropometric measures, BMD of the lumbar spine (LS) and femoral neck (FN), exercise time, selected nutrient, and energy intake were compared. The women were categorized by frame size and body mass index (BMI), with the upper range for normal weight (NW) being BMI 23.0 to 25.9 (n=30) and lower weight (LW) being BMI 16.0-19.9 (n=31). Multivariate t tests, Pearson correlations, and independent sample t tests were used for statistical analysis. Ten of 21 in the LW group, all with small frames, had varying degrees of low BMD of the LS and/or FN. The amount of exercise time was greater in the NW group. Energy and nutrient intakes did not differ significantly between groups. PMID- 12778048 TI - Self-reported vegetarianism may be a marker for college women at risk for disordered eating. AB - The purpose of this study was to determine whether differences exist in eating attitudes and behaviors of vegetarian and nonvegetarian college women. The Eating Attitudes Test (EAT) and a questionnaire were used to gather information on eating attitudes and behaviors of 143 female college students. Thirty participants were self-reported vegetarians, and 113 participants were nonvegetarians. There was no significant difference between the vegetarians and nonvegetarians in height, weight, age, or body mass index. The median EAT score of the vegetarians (16.5) was significantly higher (P<.0001) than that of the nonvegetarians (9.0). A significantly higher (P<.0001) proportion of the vegetarians (37%) compared with nonvegetarians (8%) had EAT scores greater than 30 (indicating eating disorder risk). There was no difference in supplement use or meal skipping between the two groups. In conclusion, self-reported vegetarian college women may be more likely to display disordered eating attitudes and behaviors than nonvegetarians. PMID- 12778049 TI - Position of the American Dietetic Association and Dietitians of Canada: Vegetarian diets. AB - It is the position of the American Dietetic Association and Dietitians of Canada that appropriately planned vegetarian diets are healthful, nutritionally adequate, and provide health benefits in the prevention and treatment of certain diseases. Approximately 2.5% of adults in the United States and 4% of adults in Canada follow vegetarian diets. A vegetarian diet is defined as one that does not include meat, fish, or fowl. Interest in vegetarianism appears to be increasing, with many restaurants and college foodservices offering vegetarian meals routinely. Substantial growth in sales of foods attractive to vegetarians has occurred, and these foods appear in many supermarkets. This position paper reviews the current scientific data related to key nutrients for vegetarians, including protein, iron, zinc, calcium, vitamin D, riboflavin, vitamin B-12, vitamin A, n-3 fatty acids, and iodine. A vegetarian, including vegan, diet can meet current recommendations for all of these nutrients. In some cases, use of fortified foods or supplements can be helpful in meeting recommendations for individual nutrients. Well-planned vegan and other types of vegetarian diets are appropriate for all stages of the life cycle, including during pregnancy, lactation, infancy, childhood, and adolescence. Vegetarian diets offer a number of nutritional benefits, including lower levels of saturated fat, cholesterol, and animal protein as well as higher levels of carbohydrates, fiber, magnesium, potassium, folate, and antioxidants such as vitamins C and E and phytochemicals. Vegetarians have been reported to have lower body mass indices than nonvegetarians, as well as lower rates of death from ischemic heart disease; vegetarians also show lower blood cholesterol levels; lower blood pressure; and lower rates of hypertension, type 2 diabetes, and prostate and colon cancer. Although a number of federally funded and institutional feeding programs can accommodate vegetarians, few have foods suitable for vegans at this time. Because of the variability of dietary practices among vegetarians, individual assessment of dietary intakes of vegetarians is required. Dietetics professionals have a responsibility to support and encourage those who express an interest in consuming a vegetarian diet. They can play key roles in educating vegetarian clients about food sources of specific nutrients, food purchase and preparation, and any dietary modifications that may be necessary to meet individual needs. Menu planning for vegetarians can be simplified by use of a food guide that specifies food groups and serving sizes. PMID- 12778051 TI - A new food guide for North American vegetarians. PMID- 12778052 TI - Neuroadaptations in cystine-glutamate exchange underlie cocaine relapse. AB - Repeated cocaine treatment and withdrawal produces changes in brain function thought to be involved in relapse to drug use. Withdrawal from repeated cocaine reduced in vivo extracellular glutamate in the nucleus accumbens of rats by decreasing the exchange of extracellular cystine for intracellular glutamate. In vivo restoration of cystine/glutamate exchange by intracranial perfusion of cystine or systemically administered N-acetylcysteine normalized the levels of glutamate in cocaine-treated subjects. To determine if the reduction in nonvesicular glutamate release is a mediator of relapse, we examined cocaine primed reinstatement of drug seeking after cocaine self-administration was stopped. Reinstatement was prevented by stimulating cystine/glutamate exchange with N-acetylcysteine and restoring extracellular glutamate. Thus, withdrawal from repeated cocaine increases susceptibility to relapse in part by reducing cystine/glutamate exchange, and restoring exchanger activity prevents cocaine primed drug seeking. PMID- 12778053 TI - Opsin activation as a cause of congenital night blindness. AB - Three different mutations of rhodopsin are known to cause autosomal dominant congenital night blindness in humans. Although the mutations have been studied for 10 years, the molecular mechanism of the disease is still a subject of controversy. We show here, using a transgenic Xenopus laevis model, that the photoreceptor cell desensitization that is a hallmark of the disease results from persistent signaling by constitutively active mutant opsins. PMID- 12778054 TI - Ubp3 requires a cofactor, Bre5, to specifically de-ubiquitinate the COPII protein, Sec23. AB - Ubiquitination is important for a broad array of cellular functions. Although reversal of this process, de-ubiquitination, most probably represents an important regulatory step contributing to cellular homeostasis, the specificity and properties of de-ubiquitination enzymes remain poorly understood. Here, we show that the Saccharomyces cerevisiae ubiquitin protease Ubp3 requires an additional protein, Bre5, to form an active de-ubiquitination complex that cleaves ubiquitin from specific substrates. In particular, this complex rescues Sec23p, a COPII subunit essential for the transport between the endoplasmic reticulum and the Golgi apparatus, from degradation by the proteasome. This probably contributes to maintaining and adapting a Sec23 expression level that is compatible with an efficient secretion pathway, and consequently with cell growth and viability. PMID- 12778055 TI - Development of potent monoclonal antibody auristatin conjugates for cancer therapy. AB - We describe the in vitro and in vivo properties of monoclonal antibody (mAb)-drug conjugates consisting of the potent synthetic dolastatin 10 analogs auristatin E (AE) and monomethylauristatin E (MMAE), linked to the chimeric mAbs cBR96 (specific to Lewis Y on carcinomas) and cAC10 (specific to CD30 on hematological malignancies). The linkers used for conjugate formation included an acid-labile hydrazone and protease-sensitive dipeptides, leading to uniformly substituted conjugates that efficiently released active drug in the lysosomes of antigen positive (Ag+) tumor cells. The peptide-linked mAb-valine-citrulline-MMAE and mAb phenylalanine-lysine-MMAE conjugates were much more stable in buffers and plasma than the conjugates of mAb and the hydrazone of 5-benzoylvaleric acid-AE ester (AEVB). As a result, the mAb-Val-Cit-MMAE conjugates exhibited greater in vitro specificity and lower in vivo toxicity than corresponding hydrazone conjugates. In vivo studies demonstrated that the peptide-linked conjugates induced regressions and cures of established tumor xenografts with therapeutic indices as high as 60-fold. These conjugates illustrate the importance of linker technology, drug potency and conjugation methodology in developing safe and efficacious mAb drug conjugates for cancer therapy. PMID- 12778056 TI - Engineering a mevalonate pathway in Escherichia coli for production of terpenoids. AB - Isoprenoids are the most numerous and structurally diverse family of natural products. Terpenoids, a class of isoprenoids often isolated from plants, are used as commercial flavor and fragrance compounds and antimalarial or anticancer drugs. Because plant tissue extractions typically yield low terpenoid concentrations, we sought an alternative method to produce high-value terpenoid compounds, such as the antimalarial drug artemisinin, in a microbial host. We engineered the expression of a synthetic amorpha-4,11-diene synthase gene and the mevalonate isoprenoid pathway from Saccharomyces cerevisiae in Escherichia coli. Concentrations of amorphadiene, the sesquiterpene olefin precursor to artemisinin, reached 24 microg caryophyllene equivalent/ml. Because isopentenyl and dimethylallyl pyrophosphates are the universal precursors to all isoprenoids, the strains developed in this study can serve as platform hosts for the production of any terpenoid compound for which a terpene synthase gene is available. PMID- 12778057 TI - Phase II study of docetaxel in patients with relapsed or refractory malignant lymphoma. AB - We report the activity and toxicity of docetaxel in 12 evaluable heavily pretreated patients with relapsed and refractory non-Hodgkin's lymphoma and Hodgkin's disease. In all, 42% achieved a partial response, 25% achieved stable disease. Median duration of response was 16 (10-21) weeks. The median overall survival was 70 (9-178) weeks and for responders it was 120 (22-178) weeks. One patient developed one episode of neutropenic sepsis. Docetaxel has limited activity in this group of patients. PMID- 12778058 TI - Neoadjuvant docetaxel for operable breast cancer induces a high pathological response and breast-conservation rate. AB - Docetaxel (Taxotere), alone or in combination with other anticancer agents, has proven efficacy in the first- and second-line treatment of metastatic breast cancer. This phase II study investigated the efficacy and tolerability of docetaxel as neoadjuvant chemotherapy in women with stage II-III primary operable breast cancer. Patients (n=88) were treated with six cycles of docetaxel at 100 mg m(-2) every 21 days, followed by definitive surgery and radiotherapy. After six cycles of docetaxel, the overall clinical response rate was 68.4% (CI 95%: 58.1-78.7%), including 19.0% complete remissions. Breast conservation was achieved in 72.4% of patients. A high pathological complete response (pCR) rate in breast was confirmed in 15 patients (19.8% (CI 95%: 10.8-28.8%)) on Chevallier's classification restricted to breast and in 27 patients (35.5% (CI 95%: 24.7-46.3%)) on Sataloff's classification. After a median follow-up of 30.8 months, 19 recurrences were documented with a median time to first recurrence of 17.3 months. Patients with stage III tumours had more recurrences than patients with stage II tumours (P=0.02). The principal toxicity of docetaxel is myelosuppression and 70.5% of patients developed grade III or IV neutropenia with 13.6% developing neutropenic sepsis. There was no case of severe cardiac toxicity, thrombocytopenia or any other serious adverse events. In conclusion, neoadjuvant docetaxel induces a high pCR and breast-conservation rate. Docetaxel monotherapy is a highly effective regimen that merits formal comparison with currently used combination regimens in a randomised phase III study. PMID- 12778060 TI - Concurrent chemoradiotherapy for squamous cell carcinoma of the anus using a shrinking field radiotherapy technique without a boost. AB - Chemoradiotherapy (CRT) is now widely accepted as the primary treatment modality for squamous cell cancer of the anus. While randomised trials have clearly shown CRT to be more effective than radiotherapy alone, there remains uncertainty over the optimal integration of chemotherapy and radiation. We describe a series of 50 patients treated by a site specialist gastrointestinal nonsurgical oncologist with CRT at a single UK centre. Chemotherapy comprised mitomycin C (MMC) (day 1) and 5-fluorouracil (5-FU) (days 1-4, and 29-32), concurrent with 50 Gy in 25 fractions radiation, using a two-phase shrinking field technique. A radiation boost was not planned. At a median follow-up of 48 months, 11 (22%) of the patients have failed locally, of which three have been surgically salvaged. Nine (18%) have died of anal cancer. These results are comparable with those from large randomised studies, and suggest that a two-phase shrinking field radiotherapy technique with no boost, concurrent with MMC/5-FU chemotherapy, is an effective regimen for this disease. The CRT regimen described here provides the basis for the 'control arm' of the current UK-randomised CRT trial in anal cancer (ACT2). PMID- 12778059 TI - Immunotherapy with concurrent subcutaneous GM-CSF, low-dose IL-2 and IFN-alpha in patients with progressive metastatic renal cell carcinoma. AB - The purpose of the study was to determine toxicity, efficacy and immunologic effects of concurrent subcutaneous injections of low-dose interleukin-2 (LD-IL 2), granulocyte-monocyte colony-stimulating factor (GM-CSF) and interferon-alpha 2b (IFNalpha) in progressive metastatic renal cell carcinoma. In a multicentre phase II study, 59 evaluable patients received two to six cycles of subcutaneous IL-2 (4 mIU m(-2)), GM-CSF (2.5 microg kg(-1)) and IFNalpha (5 mIU flat(-1)) for 12 days per 3 weeks with evaluation after every two cycles. Cycles were repeated in responding or stable patients. Data were analysed after a median of 30 months follow-up (range 16-48 months). In 42 patients, the immunologic response was studied and related to response and survival. The main toxicity were flu-like symptoms, malaise and transient liver enzyme elevations, necessitating IL-2 reduction to 2 mIU m(-2) in 29 patients, which should be considered the maximal tolerable dose. The response was 24% (eight out of 34, three complete response (CR), five partial response (PR)) in patients with metachronic metastases and 12% (three out of 25, 2CR, 1PR) in patients with synchronic metastases. Overall response was 19% (11 out of 59). Median survival was 9.5 months. All tested patients showed expansion and/or activation of lymphocytes, T cells and subsets, NK cells, eosinophils and monocytes. Pretreatment HLA-DR levels on monocytes and number of CD4(+)HLA-DR(+) cells correlated with response. Pretreatment number of CD4(+)HLA-DR(+) cells and postimmunotherapy levels of lymphocytes, CD3(+), CD4(+) and CD8(+) T cells, but not of NK or B cells, correlated with prolonged survival. Immunotherapy with concurrent subcutaneous GM-CSF, LD-IL-2 and IFNalpha has limited toxicity, can be given as outpatient treatment and can induce durable CR. Response and survival with this form of immunotherapy seem to be more dependent on expansion/activation of T cells than of NK cells. PMID- 12778061 TI - Seasonal variation in the diagnosis of cancer: a study based on national cancer registration in Sweden. AB - Data from the Swedish Cancer Register 1987-1996 were used to examine seasonal trends in the diagnosis of cancer. For melanomas, prostate, breast and thyroid cancer there were clear seasonal variations with reductions in the number of cases reported during the summer and December that are likely to reflect mainly hospital delays. PMID- 12778062 TI - Changes in the geographical and temporal patterns of cancer incidence among black gold miners working in South Africa, 1964-1996. AB - We describe here the results of the final 8 years of geographical and temporal data of a 33-year study of the cancer experience of 12.8 million man-years of black miners working on the gold fields of South Africa over the period 1964-96. These workers were recruited from 15 territories, the major areas during the most recent period being Lesotho (26.8%), Transkei (21.5%) and Mozambique (15%). The earliest analyses, 1964-71 and 1972-79, showed hepatocellular and oesophageal cancers to be the most frequent cancers. The final analysis, for 1989-96, however, shows marked temporal changes in the relative position of four cancers or grouped malignancies: respiratory cancer up by 236%, hepatocellular carcinoma down to 32%, oesophageal holding steady, and lymphatic system cancers up by 420%, almost certainly because of association with HIV/AIDS infection. Significant geographical variations occurring between the home areas of the miners are important, as mining operations have little to do with the cancers that develop. The causes are essentially socio-environmental rather than occupational, and this means that the rates of the major cancers in the miners are surrogate measures of the same cancers in the home areas. PMID- 12778063 TI - Childhood solid tumours in relation to population mixing around the time of birth. AB - In a retrospective cohort study of 673 787 live births in the Northern Region of England, 1975-1994, we investigated whether a higher level of population mixing around birth was a risk factor for solid tumours, by diagnostic group (Hodgkin's disease, brain and spinal tumours, neuroblastoma, other solid tumours), diagnosed during 1975-2001 under age 15 years. Logistic regression was used to relate risk to population mixing, based on (i) all movers and (ii) incomers from outside the region. Both ward and county district level analyses were performed. There was a decreased risk of brain and spinal tumours with increasing population mixing based on incomers from outside the region (OR for trend across three categories=0.79, 95% CI: 0.66-0.95, P=0.01 in the ward level analysis). Although this may be because of chance, it is consistent with a role of exposure to infection and immunological response in the aetiology of these tumours. For other tumour groups, there was no consistent evidence of an association between risk and population mixing. PMID- 12778064 TI - Familial invasive and in situ squamous cell carcinoma of the skin. AB - We used the updated nation-wide Swedish Family-Cancer Database to examine familial risks in data from 1961 to 1998 on 1252 invasive and 2474 in situ squamous cell carcinoma (SCC) of the skin among offspring, and over 10 times more among parents. In 259 families a parent and an offspring had skin SCC. The familial standardised incidence ratios (SIRs) were 2.72 for invasive and 2.40 for in situ skin cancers in offspring. Multiple skin cancers in parents were associated with increased SIRs for invasive SCC in offspring, being 2.55 for one and up to 14.93 for two invasive and two in situ cancers in parents; the corresponding in situ SCC risks were 2.28 and 7.49. The population attributable fraction for any familial skin SCC, invasive or in situ, was 4.1%. Melanoma was the only discordant tumour that was associated with invasive and in situ skin SCC. These results provide evidence that there is an underlying hereditary susceptibility for at least a part of the familial clustering for skin SCC. PMID- 12778065 TI - Dietary intakes of vitamins A, C, and E and risk of melanoma in two cohorts of women. AB - Within the two Nurses' Health Study cohorts of US women, we examined whether higher intakes of vitamin C, vitamin E, retinol, or individual tocopherols or carotenoids are associated with a lower risk of melanoma. We confirmed 414 cases of invasive melanoma among over 162,000 Caucasian women aged 25-77 years during more than 1.6 million person-years of follow-up. Diet was measured every 4 years with a food frequency questionnaire and supplement use was reported every 2 years. Several measures of sun sensitivity were assessed and included in proportional hazards models. We found that vitamins A, C, E and their individual components were not associated with a lower risk of melanoma. Only retinol intake from foods plus supplements appeared protective within a subgroup of women who were otherwise at low risk based on nondietary factors (relative risk (RR)=0.39, 95% confidence interval (CI) 0.22-0.71 for >/=1,800 vs 400 microg day(-1), P for linear trend=0.01). Contrary to expectation, we observed higher risks of melanoma with greater intakes of vitamin C from food only (RR=1.43, 95% CI 1.01-2.00 for >/=175 vs <90 mg day(-1), P for linear trend=0.05) and a significant positive dose-response with frequency of orange juice consumption (P=0.008). Further research is needed to determine whether another component in foods such as orange juice may contribute to an increase in risk. PMID- 12778066 TI - Role of paan chewing and dietary habits in cervical carcinoma in Chennai, India. AB - Non-viral factors contribute to human papillomavirus (HPV)-related cervical carcinogenesis. We investigated the role of paan chewing and dietary habits among 205 women with invasive cervical cancer (ICC) and 213 age-matched control women in Chennai, India. Odds ratios (OR) and 95% confidence intervals (CI) were computed by means of unconditional multiple regression, taking into account major correlates of ICC risk. Paan chewing showed a dose-dependent direct association with ICC (OR for >/=5 paan day(-1)=4.0; 95% CI 1.2-13.3). Among dietary habits, the highest vs lowest intake tertile for vegetables and fruit was associated with an OR of 0.5 (95% CI 0.2-1.0). Low education level and low body weight were also risk factors for ICC, but they did not account for the associations of paan chewing and low vegetable and fruit intake. In the analyses restricted to HPV positive cases and controls, the inverse association with vegetable and fruit intake was confirmed. Conversely, the adverse influence of paan chewing on ICC risk seemed to be attributable to a higher prevalence of cervical HPV infection in women who chewed. PMID- 12778067 TI - Urinary endogenous sex hormone levels and the risk of postmenopausal breast cancer. AB - To assess the relation between urinary endogenous sex steroid levels and the risk of postmenopausal breast cancer, a nested case-cohort study was conducted within a large cohort (the DOM cohort) in the Netherlands (n=9,349). Until the end of follow-up (1 January 1996), 397 postmenopausal breast cancer cases were identified and a subcohort of 424 women was then taken from all eligible women. Women using hormones were excluded, leaving 364 breast cancer cases and 382 women in the subcohort for the analyses. Concentrations of oestrone, oestradiol, testosterone, 5alpha-androstane-3alpha, 17beta-diol and creatinine were measured in first morning urine samples, which had been stored since enrolment at -20 degrees C. A Cox proportional Hazards model was used, with Barlow's adjustment for case-cohort sampling, to estimate breast cancer risk in quartiles of each of the, creatinine corrected, hormone levels, the lowest quartile being the reference group. Women with higher levels of all four of the hormones were at increased risk for postmenopausal breast cancer (highest vs lowest quartile: incidence rate ratio for oestrone (IRR(oestrone)=2.5, 95% CI: 1.6-3.8; IRR(oestradiol)=1.5, 95% CI: 1.0-2.3; IRR(testosterone)=1.6, 95% CI: 1.0-2.4; IRR(5alpha-androstane-3alpha, 17beta-diol)=1.7, 95% CI: 1.1-2.7). In conclusion, women with higher excretion levels of both oestrogens and androgens have an increased risk of breast cancer. PMID- 12778068 TI - Elevated protein kinase C alpha expression may be predictive of tamoxifen treatment failure. AB - We previously reported that stable transfection of protein kinase C alpha (PKCalpha) into T47D human breast cancer cells results in tamoxifen (TAM) resistant tumour growth. Relevance of PKCalpha expression in clinical specimens was determined by comparing PKCalpha expression in tumours from patients exhibiting disease recurrence with patients remaining disease-free following TAM treatment. Our results suggest that PKCalpha expression may predict TAM treatment failure. PMID- 12778069 TI - Absence of BRAF gene mutations in uveal melanomas in contrast to cutaneous melanomas. AB - The recent discovery of activating mutations in the BRAF gene in many cutaneous melanomas led us to screen the genomic sequence of BRAF exons 11 and 15 in a series of 48 intraocular (uveal) melanomas, together with control samples from three cutaneous melanomas and the SK-Mel-28 cell line, which has a BRAF mutation. The same mutation was detected in two-thirds of our cutaneous melanoma samples, but was not present in any uveal melanomas. This finding further underlines the distinction between uveal and cutaneous melanomas, and suggests that BRAF inhibitors are unlikely to benefit patients with uveal melanoma. PMID- 12778070 TI - Expression of high-mobility-group-protein HMGI-C mRNA in the peripheral blood is an independent poor prognostic indicator for survival in metastatic breast cancer. AB - HMGI-C belongs to the high-mobility-group-protein (HMG) family of architectural transcription factors and considerable interest has recently been shown in its expression in neoplastic tissues and apparent involvement in tumorigenesis. We could previously demonstrate an expression of HMGI-C mRNA in the peripheral blood of breast cancer patients for the first time. In this prospective study, we evaluated the independent prognostic power of HMGI-C mRNA expression in the peripheral blood of an unselected cohort of 69 patients with metastatic breast cancer using a hemi-nested reverse transcriptase polymerase chain reaction (RT PCR) followed by sequence analysis of the resulting PCR products. Multivariate analysis was performed using the Cox regression model. HMGI-C mRNA was detected in peripheral blood from 21 out of 69 (30%) patients with metastatic breast cancer. Median survival was 15.9 months in patients expressing HMGI-C, while in the group of patients without HMGI-C expression the median survival had not been reached yet after a median follow-up of 24.7 months and 85.4% were still alive in this group. Disease-specific survival was significantly worse for patients positive for HMGI-C in comparison to those not expressing HMGI-C (P=0.0001). In a multivariate regression analysis, HMGI-C remained an independent prognostic factor for overall survival (P=0.001) besides oestrogen receptor status (P=0.024) and presence of metastases in liver and lungs (P=0.029). HMGI-C expression in the peripheral blood of patients with metastatic breast cancer is a powerful independent indicator for poor overall survival and this is the first study to demonstrate its prognostic relevance in univariate and multivariate analysis. PMID- 12778071 TI - The coxsackievirus and adenovirus receptor acts as a tumour suppressor in malignant glioma cells. AB - The coxsackievirus and adenovirus receptor (CAR) is a membrane glycoprotein with a cytoplasmic domain, a transmembrane domain and an extracellular region consisting of two immunoglobulin-like domains, an amino-terminal immunoglobulin variable (IgV)-related domain (D1), which is distal to the cell surface, and a proximal IgC2 domain (D2). The coxsackievirus and adenovirus receptor has been shown to exhibit tumour suppression activity in human bladder and prostate cancer cells. In the current paper, we demonstrate that CAR is a tumour suppressor in glioma cells and that the extracellular D2 domain is not required for this inhibitory effect. This finding provides a biological basis for the observation that expression of CAR is downregulated in malignant glioma cells. This suggests that strategies to redirect adenoviruses to achieve CAR-independent infection will be necessary to realise the full potential of adenoviral vectors for cancer gene therapy. PMID- 12778072 TI - Expression of cyclin D1, D3, E, and p27 in human renal cell carcinoma analysed by tissue microarray. AB - Aberrations in the G1/S transition of the cell cycle have been observed in many malignancies and seem to be critical in the transformation process. Few studies have delineated the presence of G1/S regulatory defects and their clinical relevance in renal cell carcinoma (RCC). Therefore, we have examined the protein contents of cyclin D1, D3, E, and p27 in 218 RCCs, using tissue microarray and immunohistochemistry. The results from a subset of tumours were confirmed by Western blotting and immunohistochemical staining of regular tissue sections. Interestingly, low protein contents of cyclin D1 and p27 were associated with high nuclear grade, large tumour size, and poor prognosis for patients with conventional tumours. We further observed substantial differences in the pattern of G1/S regulatory defects between the different RCC subtypes. The majority of both conventional and papillary cases expressed p27; however, chromophobe tumours generally lacked p27 staining. In addition, conventional RCCs often expressed high cyclin D1 protein levels, while papillary RCCs exhibited high cyclin E. In summary, we have shown that G1/S regulatory defects are present in RCC and are associated with clinico-pathological parameters. The pattern of cell cycle regulatory defects also differed between RCC subtypes. PMID- 12778074 TI - FGF8 isoform b expression in human prostate cancer. AB - Overexpression of fibroblast growth factor 8 (FGF8) mRNA has been previously described in prostate cancer. Of its four isoforms, FGF8b is thought to be the most important in carcinogenesis. We hypothesised that immunodetection of FGF8b in archival prostate cancer specimens is of potential prognostic value. Using a selected cohort of prostate tumours from transurethral (n=30) and radical prostatectomies (n=59), an optimised protocol for FGF8b immunoreactivity was used to corroborate expression with clinical parameters. No expression was observed in benign prostates (n=10). In prostate cancer, immunoreactivity was localised to the malignant epithelium with weak signals in the adjacent stroma. Expression of FGF8b in stage T1 and T2 cancers were 40 and 67%, respectively. In contrast, FGF8b expression was present in 94% of T3 and 100% of T4 cancers. By histological grade, FGF8b was found in 41% of low-grade cancers (Gleason score 4-6), 60% of intermediate-grade cancers (Gleason score 7 and 92% of high-grade cancers (Gleason score 8-10). The intensity of expression was significantly associated with stage (P=0.0004) and grade (P<0.0001) of disease. We further hypothesised that FGF8b overexpression resulted from enhanced transcription and translation rather than from abnormalities involving the FGF8 gene locus. This was tested by means of fluorescent in situ hybridisation in 20 cancer specimens to map the FGF8 gene locus. FGF8 gene copy number in benign and malignant nuclei was found to be similar (2.33+/-0.57 and 2.0+/-0.81, respectively P=0.51). Based on these findings, we propose a multicentre study on cohorts of patients to further evaluate FGF8b as a potential prognostic marker in prostate cancer. PMID- 12778073 TI - Both high intratumoral microvessel density determined using CD105 antibody and elevated plasma levels of CD105 in colorectal cancer patients correlate with poor prognosis. AB - CD105 and its ligand transforming growth factor beta (TGFbeta) are modulators of angiogenesis, which drives tumour growth and metastasis. Tumour microvessel density (MVD) has proven to be an important determinant of prognosis. In this study, we have examined the prognostic value of MVD identified using Mabs to the pan-endothelial marker CD34 and to CD105 in 111 patients with colorectal cancer. The Mab to CD105 preferentially reacts with angiogenic endothelial cells. Of the 111 patients studied, 38 were alive and 73 had died of the disease. The median MVD values counted using anti-CD34 and anti-CD105 were 5 (range 1.40-9.00) and 3.10 (range 0.90-8.00), respectively. Kaplan-Meier survival analysis revealed that only MVD values obtained using CD105 Mab correlated with survival. Patients with a high MVD, above the median (3.10), showed the worst prognosis. A similar outcome was observed when MVD was divided into quartiles. In order to ascertain if this strong expression of CD105 in the tumour vasculature is reflected in patients' plasma, circulating levels of CD105, TGFbeta1 and TGFbeta3 together with the receptor-ligand complexes were quantified in patients with colorectal carcinoma and normal controls. Results showed that except for TGFbeta1, the levels of all other molecules were significantly elevated compared with controls. The levels of CD105 were positively correlated with Dukes' stages. A lower TGFbeta1 level was noted in patients with carcinoma over the controls. Furthermore, TGFbeta3 and CD105/TGFbeta3 complexes were markedly lowered in postoperative compared with preoperative plasma samples. Immunostaining revealed that TGFbeta1 was expressed in cancer cells but TGFbeta3 in the stromal cells, whereas CD105 was exclusively expressed in vascular endothelial cells of tumour blood vessels. In conclusion, this study demonstrates that MVD quantified using a Mab to CD105 is an independent prognostic parameter for survival of patients with colorectal cancer, and that plasma levels of CD105, TGFbeta1, TGFbeta3 and CD105/TGFbeta complexes may be useful markers for assessing disease progression. These data have led us to propose that quantification of these determinants may prove useful to monitor therapeutic efficacy in patients with colorectal cancer, especially those who are being treated with antiangiogenic therapies. PMID- 12778075 TI - Pimonidazole binding in C6 rat brain glioma: relation with lipid droplet detection. AB - In C6 rat brain glioma, we have investigated the relation between hypoxia and the presence of lipid droplets in the cytoplasm of viable cells adjacent to necrosis. For this purpose, rats were stereotaxically implanted with C6 cells. Experiments were carried out by the end of the tumour development. A multifluorescence staining protocol combined with digital image analysis was used to quantitatively study the spatial distribution of hypoxic cells (pimonidazole), blood perfusion (Hoechst 33342), total vascular bed (collagen type IV) and lipid droplets (Red Oil) in single frozen sections. All tumours (n=6) showed necrosis, pimonidazole binding and lipid droplets. Pimonidazole binding occurred at a mean distance of 114 microm from perfused vessels mainly around necrosis. Lipid droplets were principally located in the necrotic tissue. Some smaller droplets were also observed in part of the pimonidazole-binding cells surrounding necrosis. Hence, lipid droplets appeared only in hypoxic cells adjacent to necrosis, at an approximate distance of 181 microm from perfused vessels. In conclusion, our results show that severe hypoxic cells accumulated small lipid droplets. However, a 100% colocalisation of hypoxia and lipid droplets does not exist. Thus, lipid droplets cannot be considered as a surrogate marker of hypoxia, but rather of severe, prenecrotic hypoxia. PMID- 12778076 TI - Cooperative effects of matrix metalloproteinase and cyclooxygenase-2 inhibition on intestinal adenoma reduction. AB - Matrix metalloproteinases (MMPs) and cyclooxygenase-2 (COX-2) are expressed in both sporadic and familial adenomatous colonic polyps and tumours and have been independently shown to play causal roles in intestinal tumour formation in mouse models of colon cancer. The apparent roles of these enzymes in intestinal tumorigenesis led us to examine, in the Min mouse model of colon cancer, if selective COX-2 and MMP inhibitors provide additive or synergistic therapeutic benefits in intestinal tumour prevention. The broad-spectrum MMP inhibitor (A 177430; MMPI) and the selective COX-2 inhibitor (A-285969; COX-2I) both showed dose-dependent inhibition of the number of adenomas in Min mice. Using suboptimal doses, the MMPI reduced tumour multiplicity by 32%, the COX-2I by 48% and, both agents in combination resulted in a 67% decrease compared to control demonstrating a cooperative effect on intestinal tumorigenesis. Apoptosis, proliferation, and angiogenesis were assayed in tumors from each treatment group. These agents in combination allowed for a lowered dosage to be administered to achieve significant biological effects. Clinically, this could potentially reduce side effects associated with currently used MMP and COX-2 inhibitors. Together, these compounds could represent an easily tolerated chemopreventive approach. PMID- 12778077 TI - Mechanism of IL-12 mediated alterations in tumour blood vessel morphology: analysis using whole-tissue mounts. AB - New blood vessel formation within tumours is a critical feature for tumour growth. A major limitation in understanding this complex process has been the inability to visualise and analyse vessel formation. Here, we report on the development of a whole-tissue mount technique that allows visualisation of vessel structure. Mice expressing green fluorescent protein (GFP) made it possible to easily see GFP(+) vessels within non-GFP-expressing B16 melanoma tumours. The small fragments of tumour used in this technique were also effectively stained with fluorescent probe-conjugated antibodies, allowing characterisation of the vessels based on surface marker phenotype. The vessels within tumour tissue were much more irregular and tortuous compared to those within surrounding normal muscle. B16 tumours stably transfected with the genes for IL-12 were used to assess the effects of this cytokine on tumour growth and vessel formation. The IL 12-expressing tumours grew more slowly and had much smaller blood vessels than the large, webbed vessels characteristic of the parental tumours, effects that were dependent on interferon gamma (IFN-gamma). Vessels in the parental tumours were found to express VEGFR-3, the receptor for VEGF-C and VEGF-D. Expression of this receptor by the endothelial cells of the blood vessels was lost in the cytokine expressing tumours, thus suggesting a mechanism for the antiangiogenic effects of IL-12. The combination of the whole mount technique and the GFP transgenic mice provides a powerful method for visualising tumour vasculature and characterising the effects of agents such as cytokines. PMID- 12778078 TI - Effects of light fractionation and different fluence rates on photodynamic therapy with 5-aminolaevulinic acid in vivo. AB - To improve efficacy of photodynamic therapy (PDT) with intravenously administered 5-aminolaevulinic acid (ALA) fractionating the light dose or reducing the light intensity may be a possibility. Therefore, Syrian Golden hamsters were fitted with dorsal skinfold chambers containing an amelanotic melanoma (n=26). PDT was performed (100 mW cm(-2), 100 J cm(-2), continuously or fractionated, and 25 mW cm(-2), 100 J cm(-2); continuously or fractionated) using an incoherent light source following i.v. application of ALA. Following fractionated irradiation, the light was paused after 20 J cm(-2) for 15 min. Prior to and up to 24 h after PDT tissue, pO(2) was measured using luminescence lifetime imaging. The efficacy was evaluated by measuring the tumour volume of amelanotic melanoma cells grown subcutaneously in the back of Syrian Golden hamsters (n=36). Only high-dose PDT resulted in a significant decrease of pO(2). Irrespective of the mode of irradiation only high-dose PDT induced complete remission of all tumours (13 out of 13). It could be shown that low-dose PDT failed to induce a significant decrease of pO(2). No significant effect of fractionated irradiation was shown regarding the therapeutic efficacy 28 days after PDT. Thus performing a fractionated PDT with ALA or reducing the light intensity seems not to be successful in clinical PDT according to the present data. PMID- 12778079 TI - Cell adhesion to the extracellular matrix protein fibronectin modulates radiation dependent G2 phase arrest involving integrin-linked kinase (ILK) and glycogen synthase kinase-3beta (GSK-3beta) in vitro. AB - Cell adhesion to extracellular matrix (ECM) is thought to confer resistance against cell-damaging agents, that is, drugs and radiation, in tumour and normal cells in vitro. The dependence of cell survival on beta1-integrin-linked kinase (ILK), protein kinase Balpha/Akt (PKBalpha/Akt) and glycogen synthase kinase 3beta (GSK-3beta) activity, which participate in beta1-integrin signalling and cell cycle progression was investigated as a function of radiation exposure. Colony-formation assays on polystyrene, fibronectin (FN), laminin (LA), bovine serum albumin (BSA) or poly-L-lysine (poly-L) (0-8 Gy), kinase assays, flow cytometric DNA and annexin-V analysis and immunoblotting were performed in nonirradiated and irradiated (2 or 6 Gy) A549 human lung cancer cells and CCD32 normal human lung fibroblasts. Cell contact to FN in contrast to polystyrene elevated basal ILK, PKBalpha/Akt and GSK-3beta kinase activities in A549 and CCD32 cells, as well as the basal amount of A549 G2 phase cells. Irradiation on FN or LA as compared to polystyrene, BSA or poly-L significantly improved cell survival. Following irradiation, kinase activities were stimulated strongly on polystyrene but showed to be less prominent on FN, which was because of the FN related basal induction. Following irradiation, FN compared to polystyrene enlarged and prolonged G2 arrest in both the cell lines. For the analysis of phosphatidylinositol-3 kinase (PI3-K) dependence of protein kinases and cell cycle transition, the PI3-K inhibitors LY294002 and wortmannin were used showing decreased kinase activities, antiproliferative and radiation-dependent G2 accumulation-abrogating effects accompanied by downregulation of cyclin D1 and phospho-pRb in cells attached to polystyrene. Fibronectin partly abrogated these effects PI3-K-independently. These findings suggest a novel pathway that makes direct phosphorylation of GSK-3beta by ILK feasible after irradiation. Conclusively, the data indicate that ILK, PKBalpha/Akt and GSK-3beta are involved in modulations of the cell cycle after irradiation. These interactions are strictly dependent on ECM components in a cell line-specific manner. Our findings provide molecular insights into mechanisms likely to be important for ECM dependent cell survival and cellular radioresistance as well as tumour growth. PMID- 12778080 TI - Age-related difference in susceptibility of Apc(Min/+) mice towards the chemopreventive efficacy of dietary aspirin and curcumin. AB - The nonsteroidal anti-inflammatory drug aspirin and the spice curcumin retard adenoma formation when administered long-term to Apc(Min/+) mice, a model of human familial adenomatous polyposis coli. Both agents interfere with cyclooxygenase activity. When aspirin is administered to Apc(Min/+) mice only postweaning, but not before, it is inefficacious, while curcumin given postweaning is active. Here the hypothesis was tested that dietary aspirin (0.05%) or curcumin (0.2%) prevent or delay adenoma formation in offsprings when administered to Apc(Min/+) mothers and up to the end of weaning, but not afterwards. Whereas curcumin was without effect when administered in this way, aspirin reduced numbers of intestinal adenomas by 21%. When aspirin given up to the end of weaning was combined with curcumin administered from the end of weaning for the rest of the animals' lifetime, intestinal adenoma numbers were reduced by 38%. The combination was not superior to intervention postweaning with curcumin alone. These results show that aspirin exerts chemopreventive activity in the Apc(Min/+) mouse during tumour initiation/early promotion, while curcumin is efficacious when given at a later stage of carcinogenic progression. Thus, the results suggest that in this mouse model aspirin and curcumin act during different 'windows' of neoplastic development. PMID- 12778081 TI - Antiangiogenic properties of selected ruthenium(III) complexes that are nitric oxide scavengers. AB - The nitric oxide synthase (NOS) pathway has been clearly demonstrated to regulate angiogenesis. Increased levels of NO correlate with tumour growth and spreading in different experimental and human cancers. Drugs interfering with the NOS pathway may be useful in angiogenesis-dependent tumours. The aim of this study was to pharmacologically characterise certain ruthenium-based compounds, namely NAMI-A, KP1339, and RuEDTA, as potential NO scavengers to be used as antiangiogenic/antitumour agents. NAMI-A, KP1339 and RuEDTA were able to bind tightly and inactivate free NO in solution. Formation of ruthenium-NO adducts was documented by electronic absorption, FT-IR spectroscopy and (1)H-NMR. Pretreatment of rabbit aorta rings with NAMI-A, KP1339 or RuEDTA reduced endothelium-dependent vasorelaxation elicited by acetylcholine. This effect was reversed by 8-Br-cGMP. The key steps of angiogenesis, endothelial cell proliferation and migration stimulated by vascular endothelial growth factor (VEGF) or NO donor drugs, were blocked by NAMI-A, KP1339 and RuEDTA, these compounds being devoid of any cytotoxic activity. When tested in vivo, NAMI-A inhibited angiogenesis induced by VEGF. It is likely that the antitumour properties previously observed for ruthenium-based NO scavengers, such as NAMI-A, are related to their NO-related antiangiogenic properties. PMID- 12778082 TI - Gene therapy for bladder cancer using E1B-55 kD-deleted adenovirus in combination with adenoviral vector encoding plasminogen kringles 1-5. AB - Mutations or loss of heterozygosity of p53 are detected in approximately 50% of bladder cancers. E1B-55 kD-deleted adenovirus has been shown to kill tumour cells with defective p53 function while sparing normal cells. Here, we examined the cytolytic effect and replication of E1B-55 kD-deleted adenovirus, designated Ad5WS1, on human bladder cancer cell lines with various p53 status. Ad5WS1 caused more severe cytolytic effect and replicated more efficiently in J82 and TCC-SUP bladder cancer cells carrying mutant p53 compared with TSGH-8301 and BFTC-905 bladder cancer cells retaining wild-type p53. Introduction of dominant negative p53 into BFTC-905 cells rendered them more susceptible to Ad5WS1-induced cytolysis. Furthermore, cells susceptible to lysis caused by Ad5WS1 were not attributable to their greater infectability by adenovirus. Finally, Ad5WS1 suppressed the growth of TCC-SUP bladder tumour xenografts, which could be augmented when combined with replication-defective adenoviral vector encoding kringles 1-5 of plasminogen (K1-5), an angiogenic inhibitor. Taken together, our results show that E1B-55 kD-deleted adenovirus replicates and hence lyses bladder cancer cells with mutant p53 much more efficient than those with wild-type p53. Thus, E1B-deleted adenovirus may have therapeutic potential, especially in combination with adenoviral vector expressing K1-5, for the treatment of bladder cancer. PMID- 12778084 TI - Expectations. PMID- 12778083 TI - Smoking and breast cancer. PMID- 12778086 TI - Fruit juice and dried fruit--healthy choices or not? PMID- 12778085 TI - Warfarin and extraction. PMID- 12778087 TI - Watching and waiting. PMID- 12778088 TI - Web-based learning. PMID- 12778089 TI - Antibacterial prescribing and warfarin: a review. PMID- 12778090 TI - Interdisciplinary management of hypodontia: oral surgery. PMID- 12778091 TI - Xylitol and caries prevention--is it a magic bullet? PMID- 12778099 TI - Evaluation of gustatory function in postmenopausal women. AB - OBJECTIVE: The aim of this study is to evaluate the gustatory function in postmenopausal women. SUBJECTS AND METHODS: Whole mouth taste test and spatial taste test were administered to 20 postmenopausal women (Group 1) and 20 age matched male subjects (Group 2). Further investigations were carried out for subjective assessment of taste alteration and nutritional differences between premenopausal and postmenopausal periods. RESULTS: There was a significant reduction in sucrose perception and in palatal sensitivity in postmenopausal women (p<0.05). Only 7(35%) of the female patients noticed alteration in taste perception during the postmenopausal period. Alteration in dietary habits after menopause was reported by 9 (45%) patients. CONCLUSION: This study demonstrates that taste perception of the tongue does not change, but that there is a disorder in palatal perception in postmenopausal women. Furthermore, there is tendency to preference of sweeter foods during menopause. PMID- 12778100 TI - Propofol target-controlled infusions for sedation--a safe technique for the non anaesthetist? AB - OBJECTIVE: As an alternative to general anaesthesia for dentistry.attention has been focused on other, apparently safer, methods of pain and anxiety control. We have undertaken a study to evaluate the safety and efficacy of intravenous sedation using propofol target-controlled infusions. DESIGN: We describe a retrospective observational analysis of propofol conscious sedation as an adjunct to local anaesthesia for patients undergoing simple or surgical exodontia. All the patients were assessed,selected and treated according to standardized hospital sedation protocols. Experienced anaesthetists used a standard regimen, with ECG, pulse oximetry and non-invasive blood pressure monitoring. A standard sedation record was completed for each patient. The initial target plasma propofol concentration was set at 1.5 microg ml(-1), adjusted thereafter to achieve the desired level of sedation. Any adverse events were recorded. SETTING: Birmingham Dental Hospital. SUBJECTS: Three hundred consecutive sedation episodes in adult dental phobic patients requiring exodontia under local anaesthesia. RESULTS: Sedation and treatment were satisfactorily completed in 297 patients. The mean target propofol concentration required was 2.1 microg ml(-1), (SD = 0.47 microg ml(-1), range 1 - 4 microg ml(-1)). General anaesthesia occurred in two cases (target concentration (TC) 2 microg ml(-1)), over sedation in 11 cases(TC 1 - 3 microg ml(-1)), and transient hypoxaemia in 7 cases (TC 1.8 - 2.5 microg ml( 1)). CONCLUSION: Intravenous sedation with target-controlled propofol infusions is effective. However, significant adverse effects may occur at target concentrations below 2.1 microg ml(-1). Only anaesthetists working in an appropriate environment should practise this technique. PMID- 12778101 TI - Does oral contraceptive use affect the incidence of complications after extraction of a mandibular third molar? AB - OBJECTIVE: This study investigated whether oral contraceptive use affects the incidence of complications (pain, trismus, dry socket) in women undergoing removal of impacted mandibular third molars. PATIENTS AND METHOD: Two hundred and sixty seven women, aged 17 - 45 years, underwent removal of an impacted mandibular third molar. Eighty seven of the women were regular users of oral contraceptives. All patients were evaluated for postoperative pain, trismus and dry socket (localized alveolar osteitis). RESULTS: Mean trismus values (measured as maximum interincisal distance) were similar in the two groups of patients. Postoperative pain was significantly more frequent among women taking contraceptives, both on day 1 (30% of women taking contraceptives used analgesics, versus 11% of women not taking contraceptives, p < 0.001) and on day 5 (14% versus 5%, p = 0.024). Similarly, dry socket occurred more frequently among women taking contraceptives than among women not taking contraceptives (11% versus 4%, p = 0.017). CONCLUSIONS: The results of this study support the view that oral contraceptive use favours the appearance of dry socket and postoperative pain after extraction, but has no effect on trismus. PMID- 12778114 TI - Structural basis for negative regulation of hypoxia-inducible factor-1alpha by CITED2. AB - Expression of hypoxia-responsive genes is mediated by the heterodimeric transcription factor hypoxia-inducible factor-1 (HIF-1) in complex with the p300/CREB-binding protein (p300/CBP) transcriptional coactivator. The protein CITED2, which binds p300/CBP, is thought to be a negative regulator of HIF-1 transactivation. We show that the CITED2 transactivation domain (TAD) disrupts a complex of the HIF-1alpha C-terminal TAD (C-TAD) and the cysteine-histidine-rich 1 (CH1) domain of p300/CBP by binding CH1 with high affinity. The high-resolution solution structure of the CITED2 TAD-p300 CH1 complex shows that the CITED2 TAD, like the HIF-1alpha C-TAD, folds on a helical, Zn2+-containing CH1 scaffold. The CITED2 TAD binds a different, more extensive surface of CH1 than does the HIF 1alpha C-TAD. However, a conserved 'LPXL' sequence motif in CITED2 and HIF-1alpha interacts with an overlapping binding site on CH1. Mutation of the LPEL sequence in full-length CITED2 abolishes p300 binding in vivo. These findings reveal that CITED2 regulates HIF-1 by competing for a hot spot on the p300 CH1 domain. PMID- 12778115 TI - Bacteriophage phi29 scaffolding protein gp7 before and after prohead assembly. AB - Three-dimensional structures of the double-stranded DNA bacteriophage phi29 scaffolding protein (gp7) before and after prohead assembly have been determined at resolutions of 2.2 and 2.8 A, respectively. Both structures are dimers that resemble arrows, with a four-helix bundle composing the arrowhead and a coiled coil forming the tail. The structural resemblance of gp7 to the yeast transcription factor GCN4 suggests a DNA-binding function that was confirmed by native gel electrophoresis. DNA binding to gp7 may have a role in mediating the structural transition from prohead to mature virus and scaffold release. A cryo EM analysis indicates that gp7 is arranged inside the capsid as a series of concentric shells. The position of the higher density features in these shells correlates with the positions of hexamers in the equatorial region of the capsid, suggesting that gp7 may regulate formation of the prolate head through interactions with these hexamers. PMID- 12778116 TI - Exploring the consequences of the previous trial. PMID- 12778117 TI - Chemokine receptors: signposts to brain development and disease. PMID- 12778118 TI - Extracellular matrix molecules and synaptic plasticity. PMID- 12778119 TI - Looking into the functional architecture of the brain with diffusion MRI. PMID- 12778120 TI - Roles and rules of kainate receptors in synaptic transmission. PMID- 12778121 TI - Reelin and brain development. PMID- 12778122 TI - Neural foundations of logical and mathematical cognition. PMID- 12778123 TI - Molecular views of recombination proteins and their control. AB - The efficient repair of double-strand breaks in DNA is critical for the maintenance of genome stability and cell survival. Homologous recombination provides an efficient and faithful pathway of repair, especially in replicating cells, in which it plays a major role in tumour avoidance. Many of the enzymes that are involved in recombination have been isolated, and the details of this pathway are now being unravelled at the molecular level. PMID- 12778124 TI - Rocks: multifunctional kinases in cell behaviour. AB - ROCKs, or Rho kinases, are serine/threonine kinases that are involved in many aspects of cell motility, from smooth-muscle contraction to cell migration and neurite outgrowth. Recent experiments have defined new functions of ROCKs in cells, including centrosome positioning and cell-size regulation, which might contribute to various physiological and pathological states. PMID- 12778125 TI - Killing the messenger: short RNAs that silence gene expression. AB - Short interfering RNAs can be used to silence gene expression in a sequence specific manner in a process that is known as RNA interference. The application of RNA interference in mammals has the potential to allow the systematic analysis of gene expression and holds the possibility of therapeutic gene silencing. Much of the promise of RNA interference will depend on the recent advances in short RNA-based silencing technologies. PMID- 12778126 TI - A fuzzy mitochondrial fusion apparatus comes into focus. AB - Membrane fusion is fundamental to eukaryotic life. Unlike the predominant intracellular fusion machineries that fuse compartments bounded by a single membrane, the mitochondrial fusion machinery must sequentially fuse the outer and inner mitochondrial membranes. These coordinated fusion events rely on a transmembrane GTPase that is known as fuzzy onions or Fzo. Recent studies have revealed that Fzo has an evolutionarily conserved role in mitochondrial fusion, and they take the first strides in determining the molecular nature of such a role. PMID- 12778127 TI - Fibulins: a versatile family of extracellular matrix proteins. AB - Fibulins are a newly recognized family of extracellular matrix proteins. The five known members of the family share an elongated structure and many calcium-binding sites, owing to the presence of tandem arrays of epidermal growth factor-like domains. They have overlapping binding sites for several basement-membrane proteins, tropoelastin, fibrillin, fibronectin and proteoglycans, and they participate in diverse supramolecular structures. New insights into their biological roles are now emerging from studies of transgenic mice and of some inherited human diseases. PMID- 12778128 TI - When ubiquitin meets ubiquitin receptors: a signalling connection. AB - Ubiquitylation is emerging as a versatile device for controlling cellular functions. Here, we propose that monoubiquitylation is rapidly induced by signalling events and allows the establishment of protein-protein interactions between monoubiquitylated proteins and partners that contain distinct ubiquitin binding domains. We also put forward speculative models for the regulation of monoubiquitylation versus polyubiquitylation. PMID- 12778129 TI - The present view of the mechanism of protein folding. AB - We can track the positions and movements of all the atoms in small proteins as they fold and unfold by combining experimental studies with atomic-resolution molecular dynamics simulations. General principles as to how such complex architectures form so rapidly are now emerging from in-depth studies of a few proteins. PMID- 12778130 TI - Tumorigenesis and the angiogenic switch. AB - It has become evident that we cannot understand tumour growth without considering components of the stromal microenvironment, such as the vasculature. At the same time, the tumour phenotype determines the nature of the tumour vasculature. Much research is now devoted to determining the impact of angiogenesis on tumour development and progression, and the reciprocal influences of tumour products on the microvasculature. A more detailed understanding of the complex parameters that govern the interactions between the tumour and vascular compartments will help to improve anti-angiogenic strategies-- not only for cancer treatment, but also for preventing recurrence. PMID- 12778131 TI - Vasculogenic mimicry and tumour-cell plasticity: lessons from melanoma. AB - The gene-expression profile of aggressive cutaneous and uveal melanoma cells resembles that of an undifferentiated, embryonic-like cell. The plasticity of certain types of cancer cell could explain their ability to mimic the activities of endothelial cells and to participate in processes such as neovascularization and the formation of a fluid-conducting, matrix-rich meshwork. This ability has been termed 'vasculogenic mimicry'. How does vasculogenic mimicry contribute to tumour progression, and can it be targeted by therapeutic agents? PMID- 12778132 TI - Basement membranes: structure, assembly and role in tumour angiogenesis. AB - In recent years, the basement membrane (BM)--a specialized form of extracellular matrix (ECM)--has been recognized as an important regulator of cell behaviour, rather than just a structural feature of tissues. The BM mediates tissue compartmentalization and sends signals to epithelial cells about the external microenvironment. The BM is also an important structural and functional component of blood vessels, constituting an extracellular microenvironment sensor for endothelial cells and pericytes. Vascular BM components have recently been found to be involved in the regulation of tumour angiogenesis, making them attractive candidate targets for potential cancer therapies. PMID- 12778133 TI - Stem-cell hierarchy in skin cancer. AB - Tumour architecture mimics many of the features of normal tissues, with a cellular hierarchy that regulates the balance between cell renewal and cell death. Although many tumours contain cells with the characteristics of stem cells, the identity of the normal cells that acquire the first genetic hits leading to initiation of carcinogenesis has remained elusive. Identification of the primary cell of origin of cancers and the mechanisms that influence cell-fate decisions will be crucial for the development of novel non-toxic therapies that influence tumour-cell behaviour. PMID- 12778134 TI - Contribution of stem cells and differentiated cells to epidermal tumours. AB - The outer covering of the skin--the epidermis--is subject to sustained environmental assaults. As a result, many cells acquire potentially oncogenic mutations. Most cells are lost through differentiation, and only long-term epidermal residents, such as stem cells, accumulate the number of genetic hits that are necessary for tumour development. So, what genetic and environmental factors determine whether a mutant stem cell forms a tumour and what type of tumour will develop? PMID- 12778135 TI - The pathogenesis of cancer metastasis: the 'seed and soil' hypothesis revisited. AB - Researchers have been studying metastasis for more than 100 years, and only recently have we gained insight into the mechanisms by which metastatic cells arise from primary tumours and the reasons that certain tumour types tend to metastasize to specific organs. Stephen Paget's 1889 proposal that metastasis depends on cross-talk between selected cancer cells (the 'seeds') and specific organ microenvironments (the 'soil') still holds forth today. It is now known that the potential of a tumour cell to metastasize depends on its interactions with the homeostatic factors that promote tumour-cell growth, survival, angiogenesis, invasion and metastasis. How has this field developed over the past century, and what major breakthroughs are most likely to lead to effective therapeutic approaches? PMID- 12778136 TI - RAS oncogenes: the first 30 years. AB - From the pioneering work with acute transforming retroviruses to the current post genomic era, RAS genes have always been at the leading edge of signal transduction and molecular oncology. Yet, a complete understanding of RAS function and dysfunction - mainly in human cancer - is still to come. The knowledge that has accumulated since their discovery 30 years ago has, however, been remarkable, and should pave the way for not only solving the outstanding issues regarding RAS biology, but also for developing efficacious drugs that could have a significant impact on cancer treatment. PMID- 12778137 TI - Immunotherapy of established tumors using bone marrow transplantation with antigen gene--modified hematopoietic stem cells. AB - A major focus of cancer immunotherapy is to develop strategies to induce T-cell responses through presentation of tumor antigens by dendritic cells (DCs). Current vaccines are limited in their ability to efficiently transfer antigens to DCs in vivo. Ex vivo-generated DCs can be efficiently loaded with antigen but after reinjection, few DCs traffic to secondary lymphoid organs, the critical sites for antigen presentation. To enhance efficiency and durability of antigen presentation by DCs, we transduced hematopoietic stem-progenitor cells (HSCs) with a model tumor antigen and then transplanted the gene-modified cells into irradiated recipient mice, which resulted in efficient expression of the transgene in a large proportion of donor derived DCs in lymphoid organs. The combination of bone marrow transplantation (BMT) using transduced HSCs, systemic agents that generate and activate DCs, and mature T-cell infusion resulted in substantial expansion and activation of antigen-specific T cells. This tripartite strategy provided potent antigen-specific immunotherapy for an aggressive established tumor. PMID- 12778139 TI - Tapping a research-rich vein. PMID- 12778138 TI - A prion protein epitope selective for the pathologically misfolded conformation. AB - Conformational conversion of proteins in disease is likely to be accompanied by molecular surface exposure of previously sequestered amino-acid side chains. We found that induction of beta-sheet structures in recombinant prion proteins is associated with increased solvent accessibility of tyrosine. Antibodies directed against the prion protein repeat motif, tyrosine-tyrosine-arginine, recognize the pathological isoform of the prion protein but not the normal cellular isoform, as assessed by immunoprecipitation, plate capture immunoassay and flow cytometry. Antibody binding to the pathological epitope is saturable and specific, and can be created in vitro by partial denaturation of normal brain prion protein. Conformation-selective exposure of Tyr-Tyr-Arg provides a probe for the distribution and structure of pathologically misfolded prion protein, and may lead to new diagnostics and therapeutics for prion diseases. PMID- 12778141 TI - US to limit AIDS trials in developing nations. PMID- 12778142 TI - US AIDS initiative clears first hurdle. PMID- 12778143 TI - The road to research is paved with restrictions. PMID- 12778145 TI - Pediatric drug testing enters infancy. PMID- 12778148 TI - Mary-Claire King. PMID- 12778149 TI - Natural-born killers. PMID- 12778150 TI - A European Research Council for basic research. PMID- 12778153 TI - Autoimmunity: busting the atherosclerotic plaque. PMID- 12778155 TI - Pausing paw proliferation. PMID- 12778154 TI - Prime boost vaccines power up in people. PMID- 12778156 TI - Lamin mutations come of age. PMID- 12778158 TI - T cells bite the hand that feeds them. PMID- 12778157 TI - Rethinking the central causes of diabetes. PMID- 12778159 TI - This is your lung on diesel exhaust. PMID- 12778160 TI - Biological drug duo delivers one-two tumor punch. PMID- 12778161 TI - HIV hijacks dendritic cells. PMID- 12778163 TI - Angiogenesis in health and disease. AB - Blood vessels constitute the first organ in the embryo and form the largest network in our body but, sadly, are also often deadly. When dysregulated, the formation of new blood vessels contributes to numerous malignant, ischemic, inflammatory, infectious and immune disorders. Molecular insights into these processes are being generated at a rapidly increasing pace, offering new therapeutic opportunities that are currently being evaluated. PMID- 12778164 TI - Endothelial signaling during development. AB - Blood vessels perfuse all tissues in the body and mediate vital metabolic exchange between tissues and blood. Increasing evidence, however, points to a direct role for paracrine signaling between blood vessel cells and surrounding target organ cells, during embryonic development and cell differentiation. Understanding the nature of this signaling and its heterogeneity, both in the embryo and in adult tissues, may not only provide insights into mechanisms for normal developmental cell fate decisions, but could also lead to novel targeted therapeutic approaches for a variety of diseases such as heart disease, diabetes or cancer. PMID- 12778165 TI - The biology of VEGF and its receptors. AB - Vascular endothelial growth factor (VEGF) is a key regulator of physiological angiogenesis during embryogenesis, skeletal growth and reproductive functions. VEGF has also been implicated in pathological angiogenesis associated with tumors, intraocular neovascular disorders and other conditions. The biological effects of VEGF are mediated by two receptor tyrosine kinases (RTKs), VEGFR-1 and VEGFR-2, which differ considerably in signaling properties. Non-signaling co receptors also modulate VEGF RTK signaling. Currently, several VEGF inhibitors are undergoing clinical testing in several malignancies. VEGF inhibition is also being tested as a strategy for the prevention of angiogenesis, vascular leakage and visual loss in age-related macular degeneration. PMID- 12778166 TI - Regulation of angiogenesis by hypoxia: role of the HIF system. AB - The regulation of angiogenesis by hypoxia is an important component of homeostatic mechanisms that link vascular oxygen supply to metabolic demand. Molecular characterization of angiogenic pathways, identification of hypoxia inducible factor (HIF) as a key transcriptional regulator of these molecules, and the definition of the HIF hydoxylases as a family of dioxygenases that regulate HIF in accordance with oxygen availability have provided new insights into this process. Here we review these findings, and the role of HIF in developmental, adaptive and neoplastic angiogenesis. We also discuss the implications of oncogenic activation of extensive, physiologically interconnected hypoxia pathways for the tumor phenotype. PMID- 12778167 TI - Molecular regulation of vessel maturation. AB - The maturation of nascent vasculature, formed by vasculogenesis or angiogenesis, requires recruitment of mural cells, generation of an extracellular matrix and specialization of the vessel wall for structural support and regulation of vessel function. In addition, the vascular network must be organized so that all the parenchymal cells receive adequate nutrients. All of these processes are orchestrated by physical forces as well as by a constellation of ligands and receptors whose spatio-temporal patterns of expression and concentration are tightly regulated. Inappropriate levels of these physical forces or molecules produce an abnormal vasculature--a hallmark of various pathologies. Normalization of the abnormal vasculature can facilitate drug delivery to tumors and formation of a mature vasculature can help realize the promise of therapeutic angiogenesis and tissue engineering. PMID- 12778168 TI - Gene transfer as a tool to induce therapeutic vascular growth. AB - Therapeutic induction of vascular growth may provide a treatment option for those patients with myocardial or peripheral ischemia who are not suited to conventional revascularization therapies. Some lymphatic vascular disorders may also be amenable to this therapy. However, clear evidence of efficacy must be obtained from phase 2 and 3 clinical trials before these new treatments can be entered into clinical practice. Apart from the clinical applications, gene transfer aimed at stimulating or blocking vascular growth with various growth factors, cytokines, transcription factors and receptors or their antagonists is useful for analyzing the effects of those molecules on the vasculature, especially when gene targeting results in lethality or when large animal models are required. PMID- 12778169 TI - Therapeutic stem and progenitor cell transplantation for organ vascularization and regeneration. AB - Emerging evidence suggests that bone marrow-derived endothelial, hematopoietic stem and progenitor cells contribute to tissue vascularization during both embryonic and postnatal physiological processes. Recent preclinical and pioneering clinical studies have shown that introduction of bone marrow-derived endothelial and hematopoietic progenitors can restore tissue vascularization after ischemic events in limbs, retina and myocardium. Corecruitment of angiocompetent hematopoietic cells delivering specific angiogenic factors facilitates incorporation of endothelial progenitor cells (EPCs) into newly sprouting blood vessels. Identification of cellular mediators and tissue-specific chemokines, which facilitate selective recruitment of bone marrow-derived stem and progenitor cells to specific organs, will open up new avenues of research to accelerate organ vascularization and regeneration. In addition, identification of factors that promote differentiation of the progenitor cells will permit functional incorporation into neo-vessels of specific tissues while diminishing potential toxicity to other organs. In this review, we discuss the clinical potential of vascular progenitor and stem cells to restore long-lasting organ vascularization and function. PMID- 12778170 TI - Imaging of angiogenesis: from microscope to clinic. AB - Advances in imaging are transforming our understanding of angiogenesis and the evaluation of drugs that stimulate or inhibit angiogenesis in preclinical models and human disease. Vascular imaging makes it possible to quantify the number and spacing of blood vessels, measure blood flow and vascular permeability, and analyze cellular and molecular abnormalities in blood vessel walls. Microscopic methods ranging from fluorescence, confocal and multiphoton microscopy to electron microscopic imaging are particularly useful for elucidating structural and functional abnormalities of angiogenic blood vessels. Magnetic resonance imaging (MRI), computed tomography (CT), positron emission tomography (PET), ultrasonography and optical imaging provide noninvasive, functionally relevant images of angiogenesis in animals and humans. An ongoing dilemma is, however, that microscopic methods provide their highest resolution on preserved tissue specimens, whereas clinical methods give images of living tissues deep within the body but at much lower resolution and specificity and generally cannot resolve vessels of the microcirculation. Future challenges include developing new imaging methods that can bridge this resolution gap and specifically identify angiogenic vessels. Another goal is to determine which microscopic techniques are the best benchmarks for interpreting clinical images. The importance of angiogenesis in cancer, chronic inflammatory diseases, age-related macular degeneration and reversal of ischemic heart and limb disease provides incentive for meeting these challenges. PMID- 12778172 TI - Thromboxane A2 modulates interaction of dendritic cells and T cells and regulates acquired immunity. AB - Physical interaction of T cells and dendritic cells (DCs) is essential for T cell proliferation and differentiation, but it has been unclear how this interaction is regulated physiologically. Here we show that DCs produce thromboxane A2 (TXA2), whereas naive T cells express the thromboxane receptor (TP). In vitro, a TP agonist enhances random cell movement (chemokinesis) of naive but not memory T cells, impairs DC-T cell adhesion, and inhibits DC-dependent proliferation of T cells. In vivo, immune responses to foreign antigens are enhanced in TP-deficient mice, which also develop marked lymphadenopathy with age. Similar immune responses were seen in wild-type mice treated with a TP antagonist during the sensitization period. Thus, TXA2-TP signaling modulates acquired immunity by negatively regulating DC-T cell interactions. PMID- 12778173 TI - Mutations in proto-oncogene GFI1 cause human neutropenia and target ELA2. AB - Mice lacking the transcriptional repressor oncoprotein Gfi1 are unexpectedly neutropenic. We therefore screened GFI1 as a candidate for association with neutropenia in affected individuals without mutations in ELA2 (encoding neutrophil elastase), the most common cause of severe congenital neutropenia (SCN; ref. 3). We found dominant negative zinc finger mutations that disable transcriptional repressor activity. The phenotype also includes immunodeficient lymphocytes and production of a circulating population of myeloid cells that appear immature. We show by chromatin immunoprecipitation, gel shift, reporter assays and elevated expression of ELA2 in vivo in neutropenic individuals that GFI1 represses ELA2, linking these two genes in a common pathway involved in myeloid differentiation. PMID- 12778175 TI - Bioassay of 2-(Chloromethyl)Pyridine Hydrochloride for Possible Carcinogenicity (CAS No. 6959-47-3). AB - 2-(Chloromethyl)pyridine hydrochloride, an aromatic heterocycle used in a variety of syntheses, was selected for bioassay by the National Cancer Institute because of the structural similarity of this compound to 2-(a,b-dichloroethyl)-pyridine hydrochloride, a carcinogen in rats, mice, Syrian hamsters, and Mongolian gerbils. A bioassay for the possible carcinogenicity of 2-(chloromethyl)pyridine hydrochloride was conducted using Fischer 344 rats and B6C3F1 mice. 2 (Chloromethyl)pyridine hydrochloride was administered by gavage, at either of two dosages, to groups of 50 male and 50 female animals of each species, with the exception of 49 male rats in the high dose group. Twenty animals of each sex and species were placed on test as vehicle controls. The high and low dosages of 2 (chloromethyl)pyridine hydrochloride administered were, respectively, 150 and 75 mg/kg for rats and 250 and 125 mg/kg for mice. The compound was administered for 99 weeks to rats and mice. The period of compound administration was followed by an observation period of 6 weeks for rats and 5 weeks for mice. There were no significant positive associations between the dosages of 2-(chloromethyl)pyridine hydrochloride administered and mortality in rats or mice of either sex. Adequate numbers of animals in all groups survived sufficiently long to be at risk from late-developing tumors. Slight dose-related mean body weight depression was observed in mice of both sexes, indicating that the dosages of 2 (chloromethyl)pyridine hydrochloride administered tothese animals in this bioassay may have approximated the maximum tolerated concentrations. Since no distinct mean body weight depression relative to vehicle controls, no significant accelerated mortality, and no other signs of toxicity were associated with administration of 2-(chloromethyl)pyridine hydrochloride to rats, it is possible that these animals may have been able to tolerate a higher dosage. None of the statistical tests for any site in female rats or in mice of either sex indicated a significant positive association between compound administration and tumor incidence. There was a significant positive trend between the dosages administered and the incidences of subcutaneous fibromas in male rats. The Fisher exact comparisons, however, were not significant. Under the conditions of this bioassay, administration of 2-(chloromethyl)pyridine hydrochloride was not carcinogenic to Fischer 344 rats or B6C3F1 mice. Levels of Evidence of Carcinogenicity: Male Rats: Negative Female Rats: Negative Male Mice: Negative Female Mice: Negative Synonyms: 2-(Cl-methyl)pyridineHCl; 2-pyridylmethyl chloride hydrochloride; 2-picolylchloride hydrochloride PMID- 12778174 TI - AAV serotype 2 vectors preferentially integrate into active genes in mice. AB - Recombinant adeno-associated virus serotype 2 (rAAV2) is a promising vector for gene therapy because it can achieve long-term stable transgene expression in animals and human subjects after direct administration of vectors into various target tissues. In the liver, although stable transgene expression primarily results from extrachromosomal vector genomes, a series of experiments has shown that vector genomes integrate into host chromosomes in hepatocytes at a low frequency. Despite the low integration efficiency, recent reports of retroviral insertional mutagenesis in mice and two human subjects have raised concerns about the potential for rAAV2-mediated insertional mutagenesis. Here we characterize rAAV2-targeted chromosomal integration sites isolated from selected or non selected hepatocytes in vector-injected mouse livers. We document frequent chromosomal deletions of up to 2 kb at integration sites (14 of 14 integrations, 100%; most of the deletions were <0.3 kb) and preferred integration into genes (21 of 29 integrations, 72%). In addition, all of the targeted genes analyzed (20 of 20 targeted genes, 100%) were expressed in the liver. This is the first report to our knowledge on host chromosomal effects of rAAV2 integration in animals, and it provides insights into the nature of rAAV2 vector integration into chromosomes in quiescent somatic cells in animals and human subjects. PMID- 12778176 TI - Bioassay of Phenol for Possible Carcinogenicity (CAS No.108-95-2). AB - Phenol ranked 38th in production among U.S. chemicals in 1978 with annual production of 2.38 billion pounds. Approximately 90% of the phenol produced is used in the manufacture of phenolic (phenol formaldehyde) resins, caprolactam, bisphenol A, alkyl phenol, and adipic acid. The remainder of the phenol is used to produce an assortment of end products, including salicylic acid, phenacetin, dyes, metal cleaners, disinfectants, antiseptics, photographic chemicals, wood preservatives (pentachlorophenol), paints, paint and varnish removers, and agricultural chemicals (2,4-D and parathion). A bioassay of phenol to test for possible carcinogenicity was conducted by providing this substance in drinking water to F344 rats and B6C3F1 mice. Groups of 50 rats and 50 mice of each sex were given drinking water containing 2,500 or 5,000 ppm phenol for 103 weeks. As matched controls, groups of 50 rats and 50 mice of each sex received tap water. A dose-related depression in mean body weight gain occurred in rats and mice of each sex. Rats and mice given water containing phenol drank less than did the corresponding controls. A dose-related decrease in water consumption was observed for mice. An increased incidence of leukemia or lymphomas was detected in male rats and may have been associated with the administration of phenol. Although the incidence of these tumors in the low-dose group was significantly higher than that in controls, the incidence in the high-dose group was not. Thus an association with administration of phenol was not established. Under the conditions of this bioassay, phenol was not carcinogenic for either male or female F344 rats or male and female B6C3F1 mice. Levels of Evidence of Carcinogenicity: Male Rats: Negative Female Rats: Negative Male Mice: Negative Female Mice: Negative PMID- 12778177 TI - Bioassay of a Mixture of 1,2,3,6,7,8-Hexachlorodibenzo-p-dioxin and 1,2,3,7,8,9 Hexachlorodibenzo-p-dioxin (Dermal Study) for Possible Carcinogenicity (CAS No. 57653-85-7; CAS No. 19408-74-3). AB - Hexachlorodibenzo-p-dioxin (HCDD) is formed as a byproduct during the manufacture of certain chlorophenols and has been found in trichlorophenol, tetrachlorophenol, pentachlorophenol and in the chlorophenol-derived herbicides 2,4-dichlorophenoxyacetic acid (2,4-D) and 2,4,5-trichlorophenoxyacetic acid (2,4,5-T). From 1967 to 1970, the concentration of HCDD in commercial pentachlorophenol ranged from 0.03 to 38 ppm. Since then, HCDD levels in pentachlorophenol have been reduced to less than 1 ppm. A bioassay of a mixture of 1,2,3,6,7,8-and 1,2,3,7,8,9-hexachlorodibenzo-p-dioxins (HCDD) for possible carcinogenicity was conducted by dermal application of a suspension of this substance to Swiss-Webster mice. HCDD (0.01 &mgr;g) suspended in 0.1 ml acetone was applied to the backs of 30 mice of each sex 3 days per week for 104 weeks. During the first 16 weeks, doses were 0.005 &mgr;g HCDD per application. An additional 30 mice of each sex were pretreated with one application of 50 &mgr;g DMBA in 0.1 ml acetone 1 week before the initiation of the HCDD applications. As vehicle controls, 45 mice of each sex received 0.1 ml of acetone three times per week. Thirty animals of each sex served as untreated controls. Mean body weights of all test and vehicle control mice were comparable throughout the bioassay; mean body weights of untreated controls were higher than those of the test and vehicle-control groups. In male mice, the incidence of alveolar/bronchiolar carcinomas in the group administered only HCDD was significantly higher (P=0.045) than that in the vehicle-control group; however, the incidence was not significantly higher when compared with untreated controls. In female mice, the incidences of fibrosarcomas of the skin were significantly higher (P=0.044) in animals administered HCDD (both with and without pretreatment with DBMA) than in the untreated-control group; however, when the incidences were compared with those of the vehicle controls (relative risk=3.037) the results were not significant. Under the conditions of this bioassay, HCDD was not carcinogenic for male or female Swiss-Webster mice. Levels of Evidence of Carcinogenicity: Male Mice: Negative Female Mice: Negative Synonym: HCDD PMID- 12778178 TI - Carcinogenesis Bioassay of 2,3,7,8-Tetrachlorodibenzo-p-dioxin (CAS No. 1746-01 6) in Swiss-Webster Mice (Dermal Study). AB - 2,3,7,8-Tetrachlorodibenzo-p-dioxin occurs as a highly toxic impurity found in herbicides containing 2,4,5-trichlorophenoxyacetic acid (2,4,5-T) and 2,4,5-T- derivatives, as well as in other chemicals synthesized using 2,4,5 trichlorophenol. The herbicide 2,4,5-T has been marketed in the United States since 1948. Production increased sharply between 1960 and 1970 when a 1:1 mixture of 2,4,5-T and 2,4-dichlorophenoxyacetic acid (2,4-D) was used as a defoliant in Vietnam under the names of "herbicide agent orange, herbicide orange, agent orange, and orange". During this 10-year period, about 106 million pounds of 2,4,5-T were sprayed. A carcinogenesis bioassay was conducted by applying an acetone suspension of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) to the clipped backs of 30 male and female Swiss-Webster mice 3 days per week for 99 or 104 weeks. Similar groups were pretreated with 1 application of 50 &mgr;g dimethylbenzanthracene (DMBA) in 0.1 ml acetone 1 week before TCDD administration began. Female mice received 0.005 &mgr;g TCDD per application, and the male mice received 0.001 &mgr;g TCDD. As vehicle controls, 45 mice of each sex received 0.1 ml acetone three times per week. Thirty animals of each sex were used as untreated controls. Throughout the bioassay, mean body weights of the male and female mice administered TCDD, or TCDD following DMBA, were essentially the same as those of the corresponding vehicle control group. Mean body weights of dosed and vehicle control groups of males were less thanthose of the untreated control group throughout the study; for the females, mean body weights were less than the untreated controls during the first 80 weeks. In female mice, the incidences of fibrosarcoma in the integumentary system in dosed groups with TCDD were significantly (P=0.007) higher than that in the corresponding controls (2/41, 5%; 8/27, 30%). An increase in the same tumor type, although not statistically significant (P=0.084), was also observed in the male mice (3/42, 7%; 6/28, 21%). In the DMBA-TCDD experiment, failure to have included groups skin painted with only DMBA precluded interpretation of these results. Under the conditions of this bioassay, 2,3,7,8-tetrachlorodibenzo-p-dioxin applied to the skin was not carcinogenic for male Swiss-Webster mice (the increase of fibrosarcomas in the integumentary system may have been associated with the skin application of TCDD). TCDD was carcinogenic for female Swiss-Webster mice causing fibrosarcomas in the integumentary system. Levels of Evidence of Carcinogenicity: Male Mice: Equivocal Female Mice: Positive Synonyms: 2,3,7,8-TCDD; TCDD PMID- 12778179 TI - Bioassay of 2,6-Toluenediamine Dihydrochloride for Possible Carcinogenicity (CAS No. 15481-70-6). AB - 2,6-Toluenediamine is used as an intermediate in the production of dyes for furs and textiles, and of flexible polyurethane foams and elastomers. A bioassay of 2,6-toluenediamine dihydrochloride for possible carcinogenicity was conducted by feeding diets containing the test chemical to F344 rats and B6C3F1 mice. Groups of 50 rats of each sex were fed the test chemical at two doses, 250 or 500 ppm, for 103 weeks and observed for 1 additional week. Groups of 50 mice of each sex were fed the test chemical at two doses, 50 or 100 ppm, for 103 weeks and then observed for 1 additional week. Groups of 50 untreated rats and 50 untreated mice of each sex were used as matched controls. All surviving animals were killed and necropsied at 104 weeks. Weight gain depression was less than 10% for dosed groups of male rats and male and female mice, when compared with controls. Mean body weight gain was depressed 17% in low-dose female rats and 27% in high-dose female rats. Mortality was not increased in rats or mice of either sex by the test chemical. No clinical evidence indicated that mice of either sex received a maximum tolerated dose of the compound. In male rats, islet-cell adenomas of the pancreas and neoplastic nodules or carcinomas of the liver occurred in dose related trends that were significant using the Cochran-Armitage test (P=0.025 and P=0.037, respectively). The results of the Fisher exact test were not significant for either lesion. The occurrences of tumors in dosed female rats were not significantly different from those in control rats. Significant results in the negative direction were observed in the incidences of C-cell tumors of the thyroid in male rats and of fibroadenomas of the mammary gland in female rats. In male mice, in the low-dose group, lymphomas occurred at an incidence significantly higher (P=0.046) than that of the corresponding control group; however, the incidence was not significant when the Bonferroni criterion for multiple comparison was used. The occurrence of hepatocellular carcinomas in female mice was dose related, but the result of the Fisher exact test comparing the incidence in the high-dose group with that in the controls was not significant. It was concluded that, under the conditions of the bioassay, 2,6 toluenediamine dihydrochloride was not carcinogenic for male and female F344 rats or for male and female B6C3F1 mice. Levels of Evidence of Carcinogenicity: Male Rats: Negative Female Rats: Negative Male Mice: Negative Female Mice: Negative PMID- 12778180 TI - Bioassay of Selsun(R) for Possible Carcinogenicity. AB - A bioassay of Selsun(R) for possible carcinogenicity was conducted by applying this substance dermally to ICR Swiss mice. Selsun(R), an antidandruff shampoo, contains 2.5% selenium sulfide. Groups of 50 mice of each sex were exposed to 0.05 ml of 25% or 50% Selsun(R) in distilled water three times a week on a 2-x 3 cm clipped dorsal surface. Vehicle controls consisted of 50 mice of each sex that were clipped and treated with distilled water. Untreated controls consisted of 50 mice of each sex that were only clipped. Surviving mice were killed and necropsied at week 88. Mean body weights of untreated control, vehicle control, low-dose, and high-dose groups were comparable throughout the bioassay. Amyloidosis was a factor in the deaths of most animals after 1 year. In male mice, alveolar/bronchiolar carcinomas or adenomas occurred witha dose-related trend that was significant (P=0.008). The result of the Fisher exact test comparing the incidence in the high-dose group with that in the vehicle controls is also significant, but the incidence of the high-dose group, when compared with that of the untreated controls, is not significant. Under the conditions of this bioassay, dermal application of Selsun(R) was not carcinogenic for ICR Swiss mice. The study was limited, however, by the relatively short lifespan of this strain of mouse. Levels of Evidence of Carcinogenicity: Male Mice: Negative Female Mice: Negative PMID- 12778181 TI - Bioassay of a Mixture of 1,2,3,6,7,8-Hexachlorodibenzo-p-dioxin and 1,2,3,7,8,9 Hexachlorodibenzo-p-dioxin (Gavage) for Possible Carcinogenicity (CAS No. 57653 85-7,CAS No. 19408-74-3). AB - Hexachlorodibenzo-p-dioxins (HCDD) are formed during the manufacture of certain chlorophenols. They have been found in trichlorophenol, tetrachlorophenol, and pentachlorophenol and in the chlorophenol-derived herbicides, 2,4 dichlorophenoxyacetic acid (2,4-D) and 2,4,5-trichlorophenoxyacetic acid (2,4,5 T). From 1967 to 1970, the concentration of HCDD in commercial pentachlorophenol ranged from 0.03 to 38 ppm. Since then, HCDD levels in pentachlorophenol have been less than 1 ppm. A bioassay of a mixture of 1,2,3,6,7,8- and 1,2,3,7,8,9 hexachlorodibenzo-p-dioxin (HCDD) for possible carcinogenicity was conducted by administering the test material by gavage to Osborne-Mendel rats and B6C3F1 mice for 104 weeks. Fifty rats and 50 mice of each sex were administered HCDD suspended in a vehicle of 9:1 corn oil-acetate 2 days per week for 104 weeks at doses of 1.25, 2.5, or 5 &mgr;g/kg/wk for rats and male mice and 2.5, 5, or 10 &mgr;g/kg/wk for female mice. Seventy-five rats and 75 mice of each sex served as vehicle controls. In addition, one untreated control group containing 25 rats and 25 mice of each sex was present in the HCDD treatment room, and one untreated control group containing 25 rats and 25 mice of each sex was present in the vehicle control room. All surviving animals were killed at 105 to 108 weeks. In rats, a dose-related depression in mean body weight gain became evident in the males after week 68 of the bioassay and in the females after week 33. In mice, the mean body weight gain in the dosed groups was comparable with that of the vehicle control groups. No other toxic clinical signs were reported in either the rats or the mice. Administration of HCDD had no adverse effect on the survival of either species. In male rats, hepatocellular carcinomas or neoplastic nodules occurred at low incidences that were dose related (P=0.003). In a direct comparison, the incidence of these tumors in the high-dose group was higher (P=0.022) than that in the corresponding vehicle-control groups, but the Bonferroni requirement of P=0.017 for the multiple comparison of three dosed groups with a control group was not met. In female rats, hepatocellular carcinomas, adenomas, or neoplastic nodules occurred at incidences that were dose related (P<0.001), and in direct comparisons the incidences of these tumors in the mid-and high-dosed groups were significantly higher (P=0.006 and P<0.001, respectively) than those in the corresponding vehicle-control group. In male mice, hepatocellular carcinomas or adenomas occurred at incidences that were dose related (P=0.001), and in a direct comparison the incidence of these tumors in the high-dose group was significantly higher (P=0.001) than that in the corresponding vehicle-control group. In female mice, hepatocellular carcinomas or adenomas occurred at incidences that were dose-related (P=0.002), and the incidence of these tumors in the high-dose group was significantly higher (P=0.004) than that in the corresponding vehicle-control group. Complex nonneoplastic toxic liver lesions were seen in all dosed groups of rats and mice. Compound-associated hyperplastic lesions of the lung were also found in both male and female rats. Under the conditions of this bioassay, HCDD administered by gavage was carcinogenic, causing increased incidences of hepatocellular carcinomas or neoplastic nodules in female Osborne-Mendel rats and inducing hepatocellular carcinomas and adenomas in male and female B6C3F1 mice. HCDD was not demonstrated to be carcinogenic for male rats. Levels of Evidence of Carcinogenicity: Male Rats: Equivocal Female Rats: Positive Male Mice: Positive Female Mice: Positive Synonym: HCDD PMID- 12778182 TI - Bioassay of Selenium Sulfide (Dermal Study) for Possible Carcinogenicity (CAS No. 7446-34-6). AB - Selenium is an essential nutrient, and various selenium compounds have industrial and medical uses. The possible carcinogenicity of selenium sulfide (a component in shampoos) was investigated by applying a suspension of this substance to the skin of ICR Swiss mice. Groups of 50 mice of each sex were treated by applying 0.5 mg or 1.0 mg selenium sulfide three times a week for 86 weeks to a clipped 2 x 3-cm dorsal surface. The selenium sulfide was suspended in 0.05 ml saline solution containing 0.5% carboxymethylcellulose. Mean body weights of all dosed and control groups were comparable throughout the study. Amyloidosis, previously reported as a cause of death in Swiss mice, was a factor in the deaths of most treated and control mice after 1 year, andthe study was terminated after 88 weeks when the majority of animals in all dosed and control groups had died. Under the conditions of this bioassay, dermal application of selenium sulfide did not produce a carcinogenic effect in ICR Swiss mice, but the study was limited by the relatively short lifespan of this strain of mouse. Levels of Evidence of Carcinogenicity: Male Mice: Negative Female Mice: Negative PMID- 12778183 TI - Bioassay of Cinnamyl Anthranilate for Possible Carcinogenicity (CAS No. 87-29-6). AB - A bioassay of cinnamyl anthranilate (a synthetic flavoring agent) for possible carcinogenicity was conducted by administering the test chemical in feed to F344 rats and B6C3F1 mice. Groups of 50 rats and 50 mice of each sex were fed the test chemical in diets containing 15,000 or 30,000 ppm for 103 weeks and then observed for an additional 2 or 3 weeks. Controls consisted of groups of 50 untreated rats and 50 untreated mice of each sex. All surviving animals were killed and necropsied at 105 to 107 weeks. Mean body weights of the dosed male and female rats and mice were lower than those of the corresponding controls throughout the bioassay, and weight decrements were dose related. Mortality in rats and mice of either sex was not affected by administration of the test chemical. In male rats, adenocarcinomas or adenomas of the renal cortex and acinar-cell carcinomas or adenomas of the pancreas were found in low incidences in dosed rats but not in control rats. In direct comparisons with matched control groups, the incidences of these tumors were not significantly increased; however, because these tumors rarely occur spontaneously in aging F344 rats, they were considered to be related to compound administration. Similar pancreatic or renal tumors have not been detected among 634 historical-control male F344 rats at the same laboratory. In the female rats, no tumors occurred at incidences that could be clearly related to the administration of the test chemical. In both male and female mice, the incidences of hepatocellular carcinomas or adenomas were dose related (P<0.001) and significant (P99% pure) were conducted by administering the test chemical in corn oil by gavage to groups of 50 male and 50 female F344/N rats at doses of 125 or 250 mg/kg body weight and to groups of 50 male and 50 female B6C3F1 mice at doses of 250 or 500 mg/kg. (1,1,1,2-tetrachloroethane is used as a chemical intermediate in the production of trichloroethylene and tetrachloroethylene). Doses were administered five times per week for 103 weeks. Due to chemically induced toxicity, high-dose mice received the chemical for only 65 weeks. Groups of 50 rats and 50 mice of each sex received corn oil by gavage on the same dosing schedule and served as vehicle controls. The chemical produced cumulative toxic effects with signs of central nervous system involvement from week 44 forward in the chronic study, resulting in significantly lower survival of high-dose male rats (P=0.001), and possibly decreasing the incidence of late-developing tumors in this group. Mean body weights of dosed and control rats of each sex were similar. Fourteen control, 10 low-dose, and 3 high-dose male rats and 2 control, 5 low-dose, and 8 high-dose female rats were killed accidentally during the study; of these, 11 control and 7 low-dose males died apparently from heat stress during week 62 as a result of a 6 hour elevated temperature (>34 degrees C) in the animal room. Neither hepatocellular neoplastic nodules alone nor hepatocellular carcinomas alone occurred in statistically significant incidences in male rats, but the combined incidence of male rats with either hepatocellular neoplastic nodules or carcinomas occurred with a statistically significant positive trend (P<0.05) in the life table test (controls, 0/49, 0%; low-dose, 1/49, 2%; high-dose, 3/48, 6%). A single hepatocellular carcinoma occurred in the high-dose group. The combined incidence of liver tumors in the high-dose males (3/48, 6%) did not greatly exceed the historical incidences of liver tumors in groups of vehicle controls in other studies at this laboratory (5/243, 2.1%; range 0%-4%). However, reduced survival of the high-dose group in the present study may have reduced the sensitivity of this bioassay for detecting liver tumors. Mineralization of the kidney increased in a dose-related fashion in the male rats (12/48, 19/50, 26/48). Fibroadenomas in the mammary gland of female rats occurred with a statistically significant (P<0.05) increased incidence in the low-dose group as compared with the controls (6/49, 15/49, 7/46). The incidence in the high-dose group was not different than that in the controls. The combined incidence of adenomas, adenocarcinomas, and carcinomas in the pituitary gland of female rats showed a statistically significant (P<0.05) negative trend and the incidence in the high dose group was significantly (P<0.05) less than that in the controls (18/39, 16/45, 7/42). Mean body weight of high-dose mice was less than that of controls after week 20 in males and after week 40 in females. Clinical signs of central nervous system toxicity occurred at week 51 in both sexes of high-dose mice and by week 66 they were dead or moribund and were killed. Survival of low dose females was also significantly (P<0.05) less than that of controls. The maximum tolerated dose was exceeded in high-dose mice. Inflammation, necrosis, fatty metamorphosis, and hepatocytomegaly were observed in increased incidences in the livers of high-dose male and female mice. The major neoplastic histopathological effects occurred in the liver, where dose-related statistically significant (P<0.05) increases in the incidence of hepatocellular adenomas occurred in both male and female mice: vehicle controls, low-, and high-dose male mice had rates of 13% (6/48), 30% (14/46), and 42%, (21/50); corresponding percentages in female mice were 8% (4/49), 17% (8/46), and 50% (24/48). Evidence for the association between 1,1,1,2-tetrachloroethane and development of hepatocellular carcinomas in mice was limited because of poor survival in the high-dose grouporoethane and development of hepatocellular carcinomas in mice was limited because of poor survival in the high-dose groups. Nevertheless, there was an increased incidence of hepatocellular carcinomas in female mice despite the reduced survival in the dosed groups (controls, 1/49, 2%; low-dose, 5/46, 11%; high-dose, 6/48, 13%). There was no clear effect in male mice. Under the conditions of these studies, 1,1,1,2-tetrachloroethane was not demonstrated to be carcinogenic in F344/N rats, although the observed increase in the proportion of male rats with liver tumors may have been associated with the administration of 1,1,1,2-tetrachloroethane; accidental killing of 27 male and 15 female rats reduced the sensitivity of this bioassay for detecting a carcinogenic response. 1,1,1,2-Tetrachloroethane was carcinogenic for B6C3F1 mice, causing an increased proportion of female mice with hepatocellular carcinomas and an increased proportion of male and female mice with hepatocellular adenomas; the decreased survival in high-dose male and female mice compromised the ability of this bioassay to further determine the presence or absence of a carcinogenic effect and gave clear evidence that these doses were toxic. Levels of Evidence of Carcinogenicity: Male Rats: Equivocal Female Rats: Negative Male Mice: Positive Female Mice: Positive PMID- 12778200 TI - Carcinogenesis Bioassay of D-Mannitol (CAS No. 69-65-8) in F344/N Rats and B6C3F1 Mice (Feed Study). AB - A carcinogenesis bioassay of D-mannitol (98%-100% pure), a food and drug additive, was conducted by feeding diets containing 25,000 or 50,000 ppm D mannitol to groups of 50 F344/N rats and 50 B6C3F1 mice of each sex for 103 weeks. Groups of 50 rats and 50 mice of each sex served as controls. Survival and mean body weights of dosed and control male rats and of dosed and control mice of each sex were comparable. Survival of high-dose female rats was significantly higher (P<0.05) than that of the low-dose female rats. However, neither the survival of the low-dose group nor that of the high-dose group was significantly different from that of the controls. Throughout the study, mean body weight gain of dosed female rats was depressed (0.05) increase in tumor incidence occurred in low-dose female mice: lymphoma (2/48, 9/49, 6/50) and lymphoma or leukemia (7/48, 15/49, 7/50). These increases were not considered to be related to vinylidene chloride administration because similar effects were not found in the high-dose female mice or in male mice or rats. Under the conditions of this bioassay, vinylidene chloride administered by gavage was not carcinogenic for F344/N rats or B6C3F1/N mice of either sex. However, since the use of a maximum tolerated dose in this study has not been clearly demonstrated and since previously reported studies have shown that carcinogenicity is associated with inhalation exposure to vinylidene chloride, this study should not be taken as proof that the chemical is not a carcinogen. Levels of Evidence of Carcinogenicity: Male Rats: Negative Female Rats: Negative Male Mice: Negative Female Mice: Negative Synonyms: 1,1-dichloroethylene; VDC; 1,1-DCE PMID- 12778209 TI - Carcinogenesis Bioassay of Gum Arabic (CAS No. 9000-01-5) in F344 Rats and B6C3F1 Mice (Feed Study). AB - A carcinogenesis bioassay of gum arabic (81-86% pure), a widely used food stabilizer, was conducted by feeding diets containing 25,000 or 50,000 ppm of the test substance to 50 F344 rats and 50 B6C3F1 mice of each sex for 103 weeks. Groups of untreated rats and mice of each sex served as controls. Throughout most of the study, mean body weights of dosed male and female mice and of dosed male rats were comparable with those of the controls; mean body weights of the dosed female rats were slightly lower than those of the controls. No other compound related clinical signs or effects on survival were observed. Mean daily feed consumption by high-dose rats and mice of either sex was 85% to 94% that of the controls. The high dose (50,000 ppm) used in this bioassay is the maximum concentration (5%) currently used in feed studies. Statistically significant (P<0.05) increasing trends were observed for the number of female mice with hepatocellular carcinomas (1/49, 2/50, 6/50), and with total liver tumors (4/49, 2/50, 10/50). No statistically significant differences were obtained when comparing the control rates with those observed in the treated groups. These observations were not considered to be clearly associated with the dietary administration of gum arabic. Thus, no compound-related neoplastic or nonneoplastic lesions were found in rats or mice of either sex. Under the conditions of this bioassay, gum arabic was not carcinogenic for F344 rats or B6C3F1 mice of either sex. Levels of Evidence of Carcinogenicity: Male Rats: Negative Female Rats: Negative Male Mice: Negative Female Mice: Negative Synonym: gum acacia PMID- 12778210 TI - Carcinogenesis Bioassay of C.I. Solvent Yellow 14 (CAS No. 842-07-9) in F344/N Rats and B6C3F1 Mice (Feed Study). AB - A carcinogenesis bioassay of C.I. Solvent Yellow 14 (94.1% pure), a widely used monoazo dye, was conducted by feeding diets containing 250 or 500 ppm of C.I. Solvent Yellow 14 to groups of 50 F344 rats of either sex for 103 weeks. Similar groups of 50 B6C3F1 mice received diets containing 500 or 1,000 ppm of C.I. Solvent Yellow 14 for 103 weeks. Groups of 50 untreated rats and mice of either sex served as controls. Throughout the bioassay, mean body weights of dosed rats and mice were slightly lower than those of controls. No compound-related clinical signs or effects on survival were observed. Increases in nonneoplastic lesions included cardiac valve fibrosis for male and female rats, lymphoid hyperplasia of the lung for male rats, and for female rats, bile duct hyperplasia, focal atrophy of the pancreatic acinus, and nephropathy. None of these effects were observed in mice. Neoplastic nodules of the liver occurred in rats of either sex with a dose related trend that was significant (male, P<0.001; female, P=0.005), and the incidences in the high-dose groups were significantly higher than those in the controls (male: control, 5/50; low-dose, 10/50; high-dose, 30/50, P<0.001 and female: control, 2/50; low-dose, 3/49; high-dose, 10/48, P=0.011). Lymphomas or leukemias occurred in low-dose female mice at an incidence significantly (P<0.05) higher than that in the controls (12/50, 23/50, 17/50). Because of the lack of a dose-related trend and because the incidence in the high-dose group was not significant, the association between the increased incidence of hematopoietic tumors in the low-dose group and the administration of C.I. Solvent Yellow 14 is not clearly established. The incidence of lymphomas or leukemias in male mice was higher (not statistically significant) than that in the corresponding controls (5/49, 10/50, 10/50); in both low-and high-dose rats of either sex the incidence was significantly (P<0.001) lower than that in controls. Under the conditions of this bioassay, C.I. Solvent Yellow 14 was carcinogenic in male and female F344/N rats, as evidenced by increased incidences of neoplastic nodules of the liver. C.I. Solvent Yellow 14 was not carcinogenic for B6C3F1 mice of either sex. Levels of Evidence of Carcinogenicity: Male Rats: Positive Female Rats: Positive Male Mice: Negative Female Mice: Negative Synonym: 1-(phenylazo)-2-naphthol PMID- 12778211 TI - Carcinogenesis Bioassay of D & C Red No. 9 (CAS No. 5160-02-1) in F344 Rats and B6C3F1 Mice (Feed Study). AB - A carcinogenesis bioassay of D & C Red No. 9, a pigment used in topical drugs and cosmetics, was conducted by feeding diets containing 1,000 or 3,000 ppm of the test substance (89.8% pure) to groups of 50 F344 rats of either sex for 103 weeks. Similar groups of 50 B6C3F1 mice received diets containing 1,000 or 2,000 ppm of the test substance for 103 weeks. Groups of 50 untreated rats and mice of either sex served as controls. In a 13-week subchronic study, the spleens of most dosed rats were enlarged and pigment (unidentified) was present in the renal tubular epithelium. Lymphoreticular hyperplasia of thymic lymph nodes was found in 75-100% of females receiving 6,000-50,000 ppm D & C Red No. 9 and in 70-100% of male rats receiving 3,000-25,000 ppm. Hemosiderosis of the liver was observed at the high-dose levels in male and female rats. Mice receiving 1,250 ppm or more D & C Red No. 9 had congestion of the spleen and hemosiderin deposits. Thus, the selection of doses for the chronic study was based on the appearance of hemosiderosis and the incidences and severity of splenic lesions observed in the 91-day subchronic study. In the chronic study, mean body weights of dosed rats of either sex and of male mice were comparable with those of controls. After week 50, the mean body weight of high-dose female mice was lower than that of the controls. No compound-related effects on survival or clinical signs were observed for rats or mice of either sex. With the possible exception of female mice, all other dosed groups of rats or mice might have tolerated higher doses, thus a clear maximum tolerated dose may not have been utilized in this study. Splenic sarcomas (0/50, 0/50, 26/48; P<0.001) and neoplastic nodules of the liver (0/50, 6/50, 7/49; P<0.01) were observed in high-dose male rats at incidences significantly higher than those in the controls. Incidences of neoplastic nodules in the livers (1/50, 1/50, 5/50) of female rats showed a statistically significant (P<0.05) trend. Nonneoplastic splenic lesions were also observed in dosed male and female rats. Lymphocytic leukemia was observed in dosed male (10/50, 2/50, 2/50) and female (10/50, 2/50, 1/50) rats at statistically significant (P<0.05) decreased incidences, compared with controls. Adenomas or carcinomas of the preputial gland in male rats (7/50, 2/50, 0/50) occurred with a statistically significant (P<0.01) negative relationship to dose of D & C Red No. 9 (P=0.007). Under the conditions of this bioassay, D & C Red No. 9 was carcinogenic for male F344 rats causing an increased incidence of sarcomas of the spleen and a dose-related increase in neoplastic nodules of the liver. D & C Red No. 9 was not considered to be carcinogenic to female F344 rats, although the increased incidence of neoplastic nodules of the liver may have been associated with administration of the test chemical. D & C Red No. 9 was not carcinogenic for B6C3F1 mice of either sex. Levels of Evidence of Carcinogenicity: Male Rats: Positive Female Rats: Equivocal Male Mice: Negative Female Mice: Negative Synonyms: 5-chloro-2-[(2-hydroxy-1-naphthalenyl)azo]-4-methylbenzene sulfonic acid, barium salt; C.I. Pigment Red; C.I. Pigment Red 53:1; C.I. Pigment Red, barium salt PMID- 12778212 TI - Carcinogenesis Bioassay of Tara Gum (CAS No. 39300-88-4) in F344 Rats and B6C3F1 Mice (Feed Study). AB - A carcinogenesis bioassay of tara gum, a potential stabilizer for cosmetics and foods, was conducted by feeding diets containing 25,000 or 50,000 ppm of the test substance to 50 F344 rats and 50 B6C3F1 mice of either sex for 103 weeks. Groups of 50 untreated rats and mice of either sex served as controls. In the chronic bioassay, mean body weights of dosed and control rats of either sex were comparable over the course of the study. Feed consumption by low-and high-dose male rats was 92% and 95% that of the controls, and feed consumption by low-and high-dose female rats was 87% and 79% that of the controls. Mean body weights of high-dose mice of either sex were lower than those of controls; feed consumption by dosed mice was comparable with that of controls. Although the rats and mice might have been able to tolerate higher doses, 50,000 ppm (5%) is the recommended maximum concentration of a test substance mixed in feed, according to the guidelines of the Bioassay Program. No tumors were observed in increased incidences that were considered to be related to administration of tara gum to either species. Interstitial cell tumors of the testis in male rats were observed in a statistically significant (P 99% pure), a widely used flavor additive and chemical intermediate, were conducted by feeding diets containing 6,000 or 12,500 ppm of eugenol to groups of 50 female F344/N rats and by feeding diets containing 3,000 or 6,000 ppm to groups of 50 male F344/N rats and B6C3F1 mice of each sex for 103 weeks. Groups of 40 rats and 50 mice of each sex served as controls. Dose levels selected for the two year studies were based on thirteen-week (91-day) studies in which dietary concentrations for the six groups ranged from 0 to 12,500 ppm. Other than a -10% difference from controls in body weights in the 12,500 ppm male rats, no chemically related gross or histopathologic effects were observed. In the two-year studies, with the exception of the high dose female rats and female mice, final body weights of the treated groups were comparable to their respective controls. No significant differences in survival were apparent for any of the eight groups receiving eugenol and for the appropriate controls. Food consumption among groups was not different in comparison with controls-- rats: males equal to or greater than 97%, females equal to or greater than 91%; mice: males equal to or greater than 94%, females equal to or greater than 90%. There were no significant observable differences between treated and control groups of rats for either nonneoplastic (toxic) lesions or neoplasms that could be attributed to eugenol. Increases in tumor incidences were diagnosed for low dose male rats with alveolar, bronchiolar adenomas or carcinomas (combined), for C-cell adenomas of the thyroid gland in low dose female rats, and for endometrial stromal polyps of the uterus in high dose female rats. Fibroadenomas of the mammary gland were decreased in dosed groups of female rats compared with controls. None of these differences were considered to be associated with the dietary administration of eugenol. In male mice, the low dose animals had an increased incidence (P<0.05) of both hepatocellular adenomas (control, 4/50; low dose, 13/50; high dose, 10/49) and hepatocellular carcinomas (10/50, 20/50, 9/49) when compared with control animals. A significant increase in hepatic neoplasms was not observed in high dose animals. No single liver tumor type was observed in female mice with a statistically significant increased incidence. When the incidences of female mice with hepatocellular adenoma or carcinoma were combined (2/50, 7/49, 9/49), there was a dose-related positive trend and the incidence of liver neoplasms in high dose animals was higher than in controls (P<0.05). Eugenol was given in the diets of female F344/N rats (0, 0.6, or 1.25%) and of male F344/N rats and male and female B6C3F1 mice (0, 0.3, or 0.6%) for 103 weeks. Under these experimental conditions, there was no evidence of carcinogenicity observed for male or female rats. For mice there was equivocal evidence of carcinogenicity since eugenol caused increased incidences of both carcinomas and adenomas of the liver in male mice at the 3,000 ppm dietary level and because eugenol was associated with an increase in the combined incidences of hepatocellular carcinomas or adenomas in female mice. Levels of Evidence of Carcinogenicity: Male Rats: Negative Female Rats: Negative Male Mice: Equivocal Female Mice: Equivocal Synonym: 1-allyl-3-methoxy-4-hydroxybenzene PMID- 12778214 TI - Carcinogenesis Bioassay of Disperse Yellow 3 (CAS No. 2832-40-8) in F344 Rats and B6C3F1 Mice (Feed Study). AB - A carcinogenesis bioassay of C.I. Disperse Yellow 3 (87.6% dye), a textile dye, was conducted by feeding diets containing 5,000 or 10,000 ppm of the test substance to groups of 50 F344 rats of either sex for 103 weeks. Similar groups of 50 B6C3F1 mice received diets containing 2,500 or 5,000 ppm of the test substance for 103 weeks. Groups of 50 untreated rats and mice of each sex served as controls. Throughout the bioassay, mean body weights of dosed rats and mice of either sex were lower than those of the controls. Survival of dosed rats of either sex was significantly greater than that of the corresponding controls. No other compound-related clinical signs or effects on survival were observed. A significant increase in neoplastic nodules of the liver occurred in dosed male rats as compared to controls (controls 1/49, 2%; low-dose 15/50, 30%; P<0.001; high-dose, 10/50, 20%; P<0.01). No increase was observed for female rats. Stomach tumors, rare in F344 rats (10/2960, 0.3%), were found in the dosed male rats: one adenocarcinoma and a sarcoma in a high-dose male and in the low-dose group a squamous cell papilloma, fibrosarcoma, adenoma, and mucinous adenocarcinoma. The incidence of these tumors was not significantly greater than that in controls; thus, the association between the administration of C.I. Disperse Yellow 3 and the stomach tumors in male rats is not clearly established. Negative trends in the incidences of certain primary tumors in dosed rats included: decreased lymphocytic leukemia in both sexes; decreased malignant mesothelioma and C-cell carcinoma of the thyroid in males; and decreased pituitary chromophobe adenoma and endometrial stromal polyps in females. Hepatocellular adenomas occurred in dosed female mice at incidences significantly higher than that in the controls (control 0/50, 0%; low-dose 6/50, 12%, P<0.05; high-dose 12/50, 24%, P<0.001). The incidences of hepatocellular carcinomas were also higher in the dosed female mice than in the controls, but the increased incidences were not statistically significant (2/50, 4/50, 5/50). A significantly (P<0.05) lower incidence of hepatocellular adenomas was detected among low-dose (7/50, 1/49, 7/49) male mice. Alveolar/bronchiolar adenomas occurred in high-dose male mice at an incidence significantly (P%lt;0.05) higher than that in the controls (control 2/50, 4%; low dose 6/49, 12%; high-dose 9/49, 18%). However, the high-dose effect was not significant when adenomas and carcinomas were combined; the incidence among low dose female mice was significantly reduced as compared with controls. Thus, the incidence of alveolar/bronchiolar adenomas among males is not considered to be related to treatment with C.I. Disperse Yellow 3. Malignant lymphomas occurred in a dose-related (P<0.05) trend in female mice and at incidences greater (P<0.05) in the high-dose group than that in the controls (10/50; 16/50; 19/50). However, because of the range of variability in the historical incidence of this tumor and because of the lack of a similar effect in male mice or in male and female rats, this increase was not regarded as being unequivocally related to the administration of C.I. Disperse Yellow 3. Under the conditions of this bioassay, C.I. Disperse Yellow 3 was considered to be carcinogenic for male F344 rats, causing an increased incidence of neoplastic nodules of the liver; this dye was not carcinogenic for female F344 rats. In addition, the stomach tumors found in the male rats may have been induced by the administration of the test chemical. C.I. Disperse Yellow 3 was carcinogenic for female B6C3F1 mice, as evidenced by the increased incidence of hepatocellular adenomas; C.I. Disperse Yellow 3 was not carcinogenic for male B6C3F1 mice. Also, the increased incidence of malignant lymphoma in female mice may have been associated with the administration of C.I. Disperse Yellow 3. Levels of Evidence of Carcinogenicity: Male Rats: Positive Female Rats: Negative Male Mice: Negative Female Mice: Positive Synonyms: Disperse Fast Yellow 6; Acetamine Yellow CG PMID- 12778215 TI - Carcinogenesis Bioassay of Locust Bean Gum (CAS No. 9000-40-2) in F344 Rats and B6C3F1 Mice (Feed Study). AB - A carcinogenesis bioassay of locust bean gum, a widely used food stabilizer, was conducted by feeding diets containing 25,000 or 50,000 ppm of the test substance to 50 F344 rats and 50 B6C3F1 mice of either sex for 103 weeks. Groups of 50 untreated rats and mice of either sex served as controls. Mean body weights of high-and low-dose rats of either sex, of low-dose male mice, and of high-and low dose female mice were comparable with those of the controls; mean body weights of high-dose male mice were slightly lower than those of controls. No other compound related clinical signs or effects on survival were observed. Although the rats and mice might have been able to tolerate higher doses, 50,000 ppm (5%) is the recommended maximum concentration of a test chemical mixed in feed according to the guidelines of the Bioassay Program. Although alveolar/bronchiolar adenomas occurred in low-dose male mice at a significantly (P=0.017) higher incidence than that in the controls (7/50, 17/50, 11/50), no significant statistical results were obtained when the combined incidence of animals with either alveolar/bronchiolar adenomas or carcinomas was analyzed (14/50, 21/50, 14/50). Cortical adenomas in the adrenal gland of female rats occurred with a statistically significant (P=0.042) positive trend (1/50, 4/50, 6/50), but comparisons between test groups and the control group were not statistically different. Under the conditions of this bioassay, locust bean gum was not carcinogenic for male or female F344 rats or B6C3F1 mice. Levels of Evidence of Carcinogenicity: Male Rats: Negative Female Rats: Negative Male Mice: Negative Female Mice: Negative PMID- 12778216 TI - Carcinogenesis Bioassay of C.I. Acid Red 14 (CAS No. 3567-69-9) in F344/N Rats and B6C3F1 Mice (Feed Study). AB - A carcinogenesis bioassay of textile grade C.I. Acid Red 14 (67%-71% purity) was conducted by feeding diets containing 6,000 or 12,500 ppm of this dye for 103-104 weeks to groups of 50 male F344 rats, 12,500 or 25,000 ppm to groups of 50 female F344 rats, and 3,000 or 6,000 ppm to groups of 50 B6C3F1 mice of either sex. Groups of 90 untreated rats of either sex and 50 untreated mice of either sex served as controls. Throughout the study, mean body weights of dosed rats of either sex and dosed female mice were comparable with those of the controls, while the mean body weight of high-dose male mice was slightly lower than that of the controls. Fourteen male rats in the low-dose group and 2 in the high-dose group accidentally drowned between weeks 84 and 103; 56% and 60% of these groups survived to terminal kill compared with 78% of the controls. These losses may have reduced the sensitivity of the assay in male rats. Rats and mice may have tolerated higher doses, but the slight depression of mean body weight in high dose male mice and the non-neoplastic lesions observed in dosed female mice and in rats of both sexes suggest that doses administered in this study could be considered maximum tolerated doses. Endometrial stromal polyps of the uterus were observed in high-dose female rats at an incidence significantly higher (P=0.008) than that seen in the controls (controls: 9/87, 10%; low-dose: 11/50, 22%; high dose: 14/50, 28%). However, the observed incidence of this tumor in the dosed groups was similar to the historical rate in untreated female F344 rats at this laboratory (65/286, 23%; range 10%-37%). Hence, the increased incidence of this lesion is not regarded as being associated with the administration of C.I. Acid Red 14. Administration of C.I. Acid Red 14 to mice was not associated with an increased incidence of any tumor type. Under the conditions of this bioassay, C.I. Acid Red 14 was not carcinogenic for F344 rats or B6C3F1 mice of either sex. Levels of Evidence of Carcinogenicity: Male Rats: Negative Female Rats: Negative Male Mice: Negative Female Mice: Negative Synonym: 4-hydroxy-3-(4-Sulfo-1 naphthalenyl)azo-1-naphthalenesulfonic acid, disodium PMID- 12778217 TI - Carcinogenesis Bioassay of 2,6-Dichloro-p-Phenylenediamine (CAS No. 609-20-1) in F344 Rats and B6C3F1 Mice (Feed Study). AB - A carcinogenesis bioassay of 2,6-dichloro-p-phenylenediamine, a chemical intermediate, was conducted in groups of 50 F344 rats and B6C3F1 mice of either sex. Male rats were fed diets containing 1,000 or 2,000 ppm 2,6-dichloro-p phenylenediamine and female rats were fed 2,000 or 6,000 ppm for 103 weeks. Mice were fed 1,000 or 3,000 ppm of the test chemical for 103 weeks and observed for an additional 8 weeks. Controls consisted of 50 untreated rats and 50 untreated mice of each sex. Throughout the study, mean body weights of dosed rats and mice of either sex were lower than those of the corresponding controls. A dose-related weight gain depression was particularly pronounced for rats. Ectopic hepatocytes were observed at an increased incidence in the pancreas and nephrosis was observed in increased severity in dosed rats of either sex when compared with the corresponding controls. No increase in any tumor type was observed in treated male or female rats when compared to controls. Increased incidences of liver tumors were observed in mice of both sexes. In male mice, the incidence of hepatocellular adenomas exhibited a significant positive dose-related trend (P=0.002), and the increased incidence of hepatocellular adenomas was statistically significant in the high-dose group(4/50, 7/50, 15/50: P=0.005). The combined incidence of hepatocellular adenomas and carcinomas showed a significant positive dose-related trend (P=0.004) and was statistically significant in the high-dose group (16/50, 19/50, 29/50: P=0.008). In female mice, hepatocellular carcinomas exhibited a significant positive dose-related trend (P=0.025), but no single dose group had a statistically significant increased incidence of either adenomas (4/50, 4/50, 9/50; high-dose effect: P=0.12) or carcinomas (2/50, 2/50, 7/50; high-dose effect: P=0.08) alone. When the incidences of hepatocellular adenomas and carcinomas were combined (6/50, 6/50, 16/50), these data gave a positive dose-related trend (P=0.004) and were statistically significant in the high-dose group (P=0.014). Under the conditions of this bioassay, 2,6-dichloro-p phenylenediamine was carcinogenic for male and female B6C3F1 mice, causing increased incidences of combined hepatocellular adenomas and carcinomas, and for male B6C3F1 mice, causing an increased incidence of hepatocellular adenomas alone. 2,6-Dichloro-p-phenylenediamine was not carcinogenic for male or female F344 rats. Levels of Evidence of Carcinogenicity: Male Rats: Negative Female Rats: Negative Male Mice: Positive Female Mice: Positive PMID- 12778218 TI - Carcinogenesis Bioassay of Di(2-ethylhexyl)phthalate (CAS No. 117-81-7) in F344 Rats and B6C3F1 Mice (Feed Studies). AB - A bioassay of di(2-ethylhexyl)phthalate, the most commonly used plasticizer for polyvinylchloride polymers, for possible carcinogenicity was conducted by feeding diets containing 6,000 or 12,000 ppm of the test chemical to groups of 50 male and 50 female F344 rats and 3,000 or 6,000 ppm to groups of 50 male and 50 female B6C3F1 mice for 103 weeks. Controls consisted of 50 untreated rats and 50 untreated mice of either sex. Mean body weights of dosed male rats (high- and low dose), high-dose female rats, and dosed female mice (high- and low-dose) were marginally-to-moderately lower than those of the corresponding controls at the end of the chronic study, reflecting a decrease in body weight gain. Food consumption was reduced slightly in rats of either sex, whereas there was no apparent difference among the mouse groups. Female rats and male and female mice administered di(2-ethylhexyl)phthalate had significantly higher incidences of hepatocellular carcinomas than those observed in the controls (rats -- males: 1/50, 2%; 1/49, 2%; 5/49, 10%; females -- 0/50, 0%; 2/49, 4%; 8/50, 16%, P=0.003; mice -- males: 9/50, 18%; 14/48, 29%; 19/50, 38%, P=0.022; females: 0/50, 0%; 7/50, 14%; P=0.006, 17/50, 34%, P<0.001). Further, a statistically significant positive trend for hepatocellular carcinomas occurred in female rats (P=0.002) and in male (P=0.018) and female (P<0.001) mice. In addition, di(2 ethylhexyl)phthalate caused a statistically significant increased incidence of male rats with either hepatocellular carcinomas or neoplastic nodules (3/50, 6%; 6/49, 12%; 12/49, 24%; P=0.010). Degeneration of the seminiferous tubules was observed in the high-dose male rats (1/49, 2%; 2/44, 5%; 43/48, 90%) and in the high-dose male mice (1/49, 2%; 2/48 4%; 7/49, 14%). Hypertrophy of cells in the anterior pituitary was also found at increased incidences in the high-dose male rats (1/46, 2%; 0/43, 0%; 22/49, 45%). Under the conditions of this bioassay, di(2-ethylhexyl)phthalate was carcinogenic for F344 rats and B6C3F1 mice, causing increased incidences of female rats and male and female mice with hepatocellular carcinomas, and inducing an increased incidence of male rats with either hepatocellular carcinomas or neoplastic nodules. Levels of Evidence of Carcinogenicity: Male Rats: Positive Female Rats: Positive Male Mice: Positive Female Mice: Positive Synonym: DEHP PMID- 12778219 TI - Carcinogenesis Bioassay of 11-Aminoundecanoic Acid (CAS No. 2432-99-7) in F344 Rats and B6C3F1 Mice. AB - 11-Aminoundecanoic acid is the monomer used in the manufacture of the polyamide, nylon-11. Aminoundecanoic acid is synthesized through a series of reactions from ricinoleic acid isolated from castor bean oil. Nylon-11 is used in automobile parts, industrial fabrics (e.g. filter bags, work clothes, and netting), and brushes because of its resistance to vibration and shock and its stability when in contact with fuels. Nylon-11 resins are approved by the U.S. Food and Drug Administration for use on food contact films. A carcinogenesis bioassay of 11 aminoundecanoic acid was carried out by administering diets containing 7,500 or 15,000 ppm of 11-aminoundecanoic acid to F344 rats and B6C3F1 mice. Groups of 50 rats and 50 mice of either sex were administered the test chemical for 104 weeks (rats) or 103 weeks (mice). Controls consisted of 50 untreated rats and 50 untreated mice of each sex. Nonneoplastic effects included dose-related decreases in mean body weight gain and survival for male rats and for mice of each sex; a dose-related increased incidence of hyperplasia of the transitional epithelium of the kidney and urinary bladder in rats of each sex; and mineralization of the kidney in dosed mice of each sex. Neoplastic nodules of the liver in dosed male rats (control 1/50, 2%; low dose 9/50, 18%; high dose 8/50, 16%; P<0.01) and transitional-cell carcinomas of the urinary bladder in high-dose male rats (control 0/48, 0%; low dose 0/48, 0%; high dose 7/49, 14%: P<0.01) were observed at significantly increased incidences compared with controls. Malignant lymphomas occurred at a significantly (P<0.05) increased rate in low-dose male mice (control 2/50, 4%; low dose 9/50, 18%; high dose 4/50, 8%). Under the conditions of this bioassay, 11-aminoundecanoic acid was carcinogenic for male F344 rats, inducing neoplastic nodules in the liver and transitional-cell carcinomas in the urinary bladder. The test chemical was not carcinogenic for female F344 rats. No clear evidence was found for the carcinogenicity of 11-aminoundecanoic acid in B6C3F1 mice of either sex, although the increase in malignant lymphoma in male mice may have been associated with administration of 11-aminoundecanoic acid. Levels of Evidence of Carcinogenicity: Male Rats: Positive Female Rats: Negative Male Mice: Equivocal Female Mice: Negative PMID- 12778220 TI - Carcinogenesis Bioassay of Bisphenol A (CAS No. 80-05-7) in F344 Rats and B6C3F1 Mice (Feed Study). AB - A carcinogenesis bioassay of bisphenol A, an intermediate used in the manufacture of epoxy, polycarbonate, and polyester-styrene resins, was conducted by feeding diets containing 1,000 or 2,000 ppm of the test chemical to groups of 50 F344 rats of either sex, 1,000 or 5,000 ppm to groups of 50 male B6C3F1 mice, and 5,000 or 10,000 ppm to groups of 50 female B6C3F1 mice for 103 weeks. Groups of 50 rats and 50 mice of either sex served as controls. Mean body weights of rats of either sex and of high-and low-dose female mice and high-dose male mice were lower than those of the controls throughout the study. Since food consumption of dosed female rats was only 70% to 80% that of the controls throughout most of this study, reduced body weight gain was probably due to reduced food consumption. Food consumption by dosed male rats was 90% that of controls. Food consumption among all groups of mice appear to be similar. Leukemias occurred at increased incidences in dosed rats of both sexes. In male rats, the dose-related (13/50, 12/50, 23/50) trend was statistically significant (P=0.021) by a Cochran Armitage test, but neither the trend nor the increase in the high-dose group was significant by life table analyses, which adjust for survival differences among groups. The increased incidences in dosed female rats were also not statistically significant (7/50, 13/50, 12/50). Interstitial-cell tumors of the testes occurred at statistically significant incidences in low- and high-dose male rats; however, since this lesion normally occurs at a high incidence in aging F344 male rats, the increased incidence observed in this study was not considered compound related (35/49, 48/50, 46/49). In male mice, there was an increased incidence of leukemias or lymphomas (2/49, 9/50, 5/50), but this increase was not statistically significant. A compound-related increased incidence of multinucleated giant hepatocytes was observed in male mice (1/49, 41/49, 41/50), but there was no increase of liver tumors in male mice. The marginally significant increase in leukemias in male rats, along with an increase (not statistically significant) in leukemias in female rats and a marginally significant increase in the combined incidence of lymphomas and leukemias in male mice, suggests that exposure to bisphenol A may be associated with increased cancers of the hematopoietic system. A statistically significant increase in interstitial-cell tumors of the testes in male rats was also suggestive of carcinogenesis, but was not considered to be convincing evidence of a compound related effect because this lesion normally occurs at a high incidence in aging F344 rats. Under the conditions of this bioassay, there was no convincing evidence that bisphenol A was carcinogenic for F344 rats or B6C3F1 mice of either sex. Levels of Evidence of Carcinogenicity: Male Rats: Equivocal Female Rats: Equivocal Male Mice: Equivocal Female Mice: Negative Synonym: 4,4' isopropylidenediphenol PMID- 12778221 TI - Carcinogenesis Bioassay of Caprolactam (CAS No. 105-60-2) in F344 Rats and B6C3F1 Mice (Feed Study). AB - A carcinogenesis bioassay of caprolactam, a chemical intermediate used in the production of nylon 6, was conducted by feeding diets containing 3,750 or 7,500 ppm caprolactam to groups of 50 male or female F344 rats and 7,500 or 15,000 ppm to groups of 50 male or female B6C3F1 mice for 103 weeks. Control groups consisted of 50 undosed rats and 50 undosed mice of each sex. Throughout the bioassay, mean body weight gains for dosed rats and mice of either sex were decreased when compared with those of the controls. No other compound-related effects were observed. Under the conditions of this bioassay, caprolactam was not carcinogenic for F344 rats or B6C3F1 mice. Levels of Evidence of Carcinogenicity: Male Rats: Negative Female Rats: Negative Male Mice: Negative Female Mice: Negative Synonyms: aminocaproic lactam; 2-oxohexamethylenimine PMID- 12778222 TI - Carcinogenesis Bioassay of Butyl Benzyl Phthalate (CAS No. 85-68-7) in F344/N Rats and B6C3F1 Mice (Feed Study). AB - A carcinogenesis bioassay of butyl benzyl phthalate, a plasticizer for vinyl chloride plastics, was accomplished by feeding diets containing 6,000 or 12,000 ppm of the phthalate to groups of 50 F344/N rats and 50 B6C3F1 mice of each sex for 28 to 103 weeks. Mean body weights of dosed female rats and mice of each sex were lower than those of the control animals throughout most of the study. After week 14, an increasing number of dosed male rats died as a result of an unexplained internal hemorrhaging, and all surviving male rats were killed at week 29 to 30. Because of compound-related mortality, butyl benzyl phthalate was not adequately tested for carcinogenicity in male F344/N rats. Mononuclear cell leukemias occurred at a statistically significant (P=0.011) increased incidence in the high-dose group of female rats when compared with the control group and with a significantly (P=0.006) increasing trend (controls 7/49, 14%; low-dose 7/49, 14%; high-dose 18/50, 36%). The incidence in the high-dose group and the overall trend remained statistically significant (P=0.008 and P=0.019) when compared with the historical incidence for F344/N female rats with leukemia at this laboratory (77/ 399, 19%). Further, this leukoproliferation was generally characterized by splenomegaly and often by hepatomegaly. Administration of butyl benzyl phthalate was not associated with increased incidences of any type of tumor among male or female mice. Tumor rates were decreased in female rats for fibroadenomas of the mammary glands (20/49, 14/49, 9/50) and in male mice for lymphomas of the hematopoietic system (13/50, 11/49, 4/50) and for alveolar/bronchiolar adenomas or carcinomas (17/50, 11/49, 8/50). Under the conditions of this bioassay, butyl benzyl phthalate was probably carcinogenic for female F344/N rats, causing an increased incidence of mononuclear cell leukemias. The male F344/N rat study was considered inadequate for evaluation due to compound-related toxicity and early mortality. Butyl benzyl phthalate was not carcinogenic for B6C3F1 mice of either sex. Levels of Evidence of Carcinogenicity: Male Rats: Inadequate Study Female Rats: Positive Male Mice: Negative Female Mice: Negative Synonyms: BBP; benzyl butyl phthalate; phthalic acid; benzyl butyl ester; Santicizer 160 PMID- 12778223 TI - Carcinogenesis Bioassay of Di(2-ethylhexyl)adipate (CAS No. 103-23-1) in F344 Rats and B6C3F1 Mice (Feed Study). AB - Di(2-ethylhexyl)adipate is a plasticizer used to give flexibility to vinyl plastics. A carcinogenesis bioassay was conducted by feeding diets containing 12,000 or 25,000 ppm of di(2-ethylhexyl)adipate to groups of 50 male and 50 female F344 rats and 50 male and 50 female B6C3F1 mice for 103 weeks. Groups of 50 undosed rats and mice of each sex served as controls. All surviving animals were killed at 104 to 107 weeks. Mean body weights of high-dose rats and mice of either sex were lower than those of the controls throughout the study. Compound administration was not associated with tumor formation in F344 rats of either sex. Hepatocellular carcinomas or adenomas occurred in mice of both sexes in a dose-related fashion at incidences that were significantly higher for high-dose males and for low- and high-dose females than those in the controls. When compared with the incidence in historical laboratory control mice, however, the liver tumors in male mice could not be clearly related to compound administration. Under the conditions of this bioassay, di(2-ethylhexyl)adipate was not carcinogenic for F344 rats. Di(2-ethylhexyl)adipate was carcinogenic for female B6C3F1 mice, causing increased incidences of hepatocellular carcinomas, and was probably carcinogenic for male B6C3F1 mice, causing hepatocellular adenomas. Levels of Evidence of Carcinogenicity: Male Rats: Negative Female Rats: Negative Male Mice: Positive Female Mice: Positive Synonyms: bis(2 ethylhexyl)adipate; DEHA; octyl adipate; dioctyl adipate; DOA PMID- 12778224 TI - Carcinogenesis Studies of C.I. Acid Orange 10 (CAS No. 1936-15-8) in F344 Rats and B6C3F1 Mice (Feed Studies). AB - Carcinogenesis studies of 80% pure C.I. Acid Orange 10 (a monoazo textile dye) were conducted by feeding to groups of 50 male and 50 female F344/N rats diets containing 1,000 or 3,000 ppm C.I. Acid Orange 10 for 103 weeks. Groups of 50 male and 50 female B6C3F1 mice were fed diets containing 3,000 or 6,000 ppm for 103 weeks. Groups of 90 male and 90 female untreated rats and 50 male and 50 female untreated mice served as controls. Mean body weights and clinical signs of control and dosed rats and mice were comparable. Because no toxic effects or consistent weight differences were observed, the rats and mice may have been able to tolerate higher doses. In male rats with neoplastic nodules of the liver, the dose response trend was positive (P<0.05) and the incidence in the 3,000 ppm group was increased (P<0.05) compared to controls (control, 5/90, 6%; low dose, 3/50, 6%; high dose, 8/50, 16%). One male rat in the high dose group had both a neoplastic nodule and a carcinoma of the liver. This marginal increase in liver cell neoplasms may have been associated with the dietary administration of C.I. Acid Orange 10. For both dose groups of male and female rats, leukemia was significantly (P<0.05) decreased in a dose related (P<0.005) trend (male: 22/90, 24%; 4/50, 8%; 3/50, 6%; female: 16/88, 18%; 2/50, 4%; 0/50). No compound-related nonneoplastic or neoplastic lesions were observed in the female rats or in mice of either sex. For 103 weeks C.I. Acid Orange 10 was given in the diets of male and female F344/N rats (0, 0.1, or 0.3%) and of male and female B6C3F1 mice (0, 0.3%, or 0.6%). Under these conditions, there was no evidence of carcinogenicity for male and female F344/N rats or for male and female B6C3F1 mice. Levels of Evidence of Carcinogenicity: Male Rats: Negative Female Rats: Negative Male Mice: Negative Female Mice: Negative Synonym: 7-hydroxy-8-(phenylazo)-1,3 naphthalenedisulfonic acid, disodium salt PMID- 12778225 TI - Carcinogenesis Bioassay of 1,2-Dibromoethane (CAS No. 106-93-4) in F344 Rats and B6C3F1 Mice (Inhalation Study). AB - A carcinogenesis bioassay of 1,2-dibromoethane, a widely used nematocide and leaded gasoline additive, was conducted by exposing groups of 50 F344 rats and B6C3F1 mice of each sex by inhalation to concentrations of 10 or 40 ppm of the 1,2-dibromoethane for 78-103 weeks. Untreated controls consisted of 50 rats and 50 mice of each sex exposed in chambers to ambient air. Throughout the study, mean body weights of high-dose rats and high-dose mice of either sex were lower than those of the corresponding untreated controls. Survival of the high-dose rats of either sex and of the low- and high-dose female mice was significantly shorter than that in the corresponding controls. The principal cause of early death in control and dosed male mice was ascending, suppurative urinary tract infection that resulted in necrotic, ulcerative lesions around the urethral opening, chronic or suppurative cystitis (often with urinary tract obstruction), and ascending suppurative pyelonephritis. Carcinomas and adenocarcinomas of the nasal cavity were observed with significantly increased incidences (P<0.001) in high-dose rats of either sex relative to controls. The incidences of adenocarcinomas and adenomas of the nasal cavity were also significantly increased (P<0.001) in low-dose rats of either sex. Adenomatous polyps of the nasal cavity showed significantly increased incidence (P<0.001) in low-dose male rats. The combined incidence of alveolar/bronchiolar adenomas and carcinomas was statistically significant (P=0.024) for high-dose female rats. Hemangiosarcomas of the circulatory system (mainly spleen) and mesotheliomas of the tunica vaginalis occurred in high-dose male rats with significantly increased incidences (P<0.001) relative to controls. The incidence of fibroadenomas of the mammary gland was significantly elevated (P<0.001) in dosed female rats relative to controls. The incidences of alveolar/bronchiolar carcinoma and alveolar/bronchiolar adenoma were significantly increased(P<0.001) in high-dose male mice relative to controls. These tumors were also increased in high-dose female mice (P=0.007 for adenomas and P<0.001 for carcinomas). Hemangiosarcomas occurred in low- and high dose female mice at incidences significantly greater (P<0.001) than the incidence in the controls (0/50). High-dose female mice also had significantly increased incidences of subcutaneous fibrosarcomas (P<0.001) and of nasal cavity carcinomas (P=0.013). Low-dose female mice also showed a significantly increased incidence (P<0.001) of mammary gland adenocarcinomas. Exposure to 1,2-dibromoethane was also associated with hepatic necrosis and toxic nephropathy in rats of either sex, testicular degeneration in male rats, retinal degeneration in female rats, and epithelial hyperplasia of the respiratory system in mice. Under the conditions of this bioassay, 1,2-dibromoethane was carcinogenic for F344 rats, causing increased incidences of carcinomas, adenocarcinomas, adenomas of the nasal cavity, and hemangiosarcomas of the circulatory system in males and females; mesotheliomas of the tunica vaginalis and adenomatous polyps of the nasal cavity in males; and fibroadenomas of the mammary gland and alveolar/bronchiolar adenomas and carcinomas (combined) in females. 1,2-Dibromoethane was carcinogenic for B6C3F1 mice, causing alveolar/bronchiolar carcinomas and alveolar/bronchiolar adenomas in males and females; and hemangiosarcomas of the circulatory system, fibrosarcomas in the subcutaneous tissue, carcinomas of the nasal cavity, and adenocarcinomas of the mammary gland in females. Levels of Evidence of Carcinogenicity: Male Rats: Positive Female Rats: Positive Male Mice: Positive Female Mice: Positive Synonyms: ethylene dibromide; EDB; ethylene bromide PMID- 12778226 TI - Carcinogenesis Bioassay of 2,3,7,8-Tetrachlorodibenzo-p-dioxin (CAS No. 1746-01 6) in Osborne-Mendel Rats and B6C3F1 Mice (Gavage Study). AB - A carcinogenesis bioassay of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), a contaminant in several phenoxy herbicides, was conducted by administering TCDD by gavage to Osborne-Mendel rats and B6C3F1 mice for 104 weeks. Fifty rats and mice of each sex were given TCDD suspended in a vehicle of 9:1 corn oil-acetone 2 days per week for 104 weeks at doses of 0.01, 0.05, or 0.5 &mgr;g/kg/wk for rats and male mice and 0.04, 0.2, or 2.0 &mgr;g/kg/wk for female mice. Seventy-five rats and 75 mice of each sex served as vehicle controls. One untreated control group containing 25 rats and 25 mice of each sex was present in the TCDD treatment room, and one untreated control group containing 25 rats and 25 mice of each sex was present in the vehicle-control room. All surviving animals were killed at 105 to 108 weeks. In rats, a dose-related depression in mean body weight gain was observed in the males after week 55 of the bioassay and in the females after week 45. In mice, the mean body weight gain in the dosed groups was comparable to that of the vehicle-control groups. In male rats, increased incidences of follicular cell adenomas in the thyroid were dose related and were significantly (P=0.001) higher in the high-dose group than in the vehicle controls (1/69, 1%; 5/48, 10%; 6/50, 12%; 10/50, 20%). Similarly in the female rats, an increase (though not statistically significant) was seen in the high-dose group (3/73, 4%; 2/45, 4%; 1/49, 2%; 6/47, 13%). In female rats, the incidence of neoplastic nodules of the liver in the high-dose group was significantly (P=0.006) higher than that in the vehicle control group (5/75, 7%; 1/49, 2%; 3/50, 6%; 12/49, 24%). In male and female mice, incidences of hepatocellular carcinomas were dose related and the incidences in the high-dose groups were significantly (P=0.002 and 0.014, respectively) higher than those in the corresponding vehicle control groups (males: 8/73, 11%; 9/49, 18%; 9/49, 16%; 17/50, 34%; females: 1/73, 1%; 2/50, 4%; 2/48, 4%; 6/47, 13%). In female mice, follicular-cell adenomas in the thyroid occurred at dose-related incidences, and were significantly (P=0.009) higher in the high-dose groups than those in the vehicle controls (0/69, 0%; 3/50, 6%; 1/47, 2%; 5/46, 11%). Increased incidences of toxic hepatitis related to the administration of the test chemical were detected among high-dose rats and high dose mice of each sex. Under the conditions of this bioassay, 2,3,7,8 tetrachlorodibenzo-p-dioxin was carcinogenic for Osborne-Mendel rats, including follicular-cell thyroid adenomas in males and neoplastic nodules of the liver in females. TCDD was also carcinogenic for B6C3F1 mice, including hepatocellular carcinomas in male and females and follicular-cell thyroid adenomas in females. Levels of Evidence of Carcinogenicity: Male Rats: Positive Female Rats: Positive Male Mice: Positive Female Mice: Positive Synonyms: 2,3,7,8-TCDD; TCDD PMID- 12778227 TI - Carcinogenesis Bioassay of FD & C Yellow No. 6 (CAS No. 2783-94-0). AB - A carcinogenesis bioassay was conducted using groups of 50 male and 50 female F344 rats and B6C3F1 mice which were fed diets containing 12,500 or 25,000 ppm FD & C Yellow No. 6, a widely used food colorant, for 103 weeks. Groups of 90 male and 90 female rats and 50 male and 50 female mice served as undosed controls. Throughout the study, mean body weights of high-dose female rats and all low-dose groups were comparable with those of the controls, but mean body weights of high dose male rats and high-dose male and female mice were slightly lower (10% or less) than those of the controls. No compound-related neoplastic or nonneoplastic lesions were observed in the rats. The incidence of hepatocellular carcinomas in low-dose male mice was significantly higher than that in the controls, but the lack of a significant increase in high-dose males and the variability of liver tumors in B6C3F1 male mice precluded clearly relating the occurrence of these tumors to the administration of FD & C Yellow No.< >6. Under the conditions of this bioassay, there was no clear evidence of the carcinogenicity of FD & C Yellow No. 6 in F344 rats or B6C3F1 mice of either sex. Levels of Evidence of Carcinogenicity: Male Rats: Negative Female Rats: Negative Male Mice: Negative Female Mice: Negative Synonym: Sunset Yellow FCF PMID- 12778228 TI - Carcinogenesis Bioassay of Cytembena (CAS No. 21739-91-3). AB - A carcinogenesis bioassay of cytembena, a cytostatic agent, was conducted by injecting intraperitoneally 7 or 14 mg/kg into groups of 50 male and 50 female F344 rats and 12 or 24 mg/kg into groups of 50 male or 50 female B6C3F1 mice three times per week for 104 weeks. Groups of 50 rats and 50 mice of both sexes served as vehicle controls. Mean body weights of dosed and vehicle-control rats were comparable throughout the bioassay. Mean body weights of dosed and vehicle control mice were comparable for the first 73 weeks of the bioassay; mean body weight of the high dose male mice was slightly lower than that of the vehicle controls after 73 weeks, and that of the high-dose female mice was lower after week 87. In dosed male rats, mesotheliomas in the tunica vaginalis and malignant mesotheliomas in multiple organs occurred with dose-related trends and at incidences in each of the dosed groups which were significantly higher than those in the vehicle control rats. In dosed female rats, fibroadenomas in the mammary gland occurred with a dose-related trend and at a significantly higher incidence in the high-dose group than in vehicle control rats. Under the conditions of this bioassay, cytembena was carcinogenic for male and female F344 rats, causing increased incidences of mesotheliomas in the tunica vaginalis and in multiple organs of males and fibroadenomas in the mammary gland of females. Cytembena was not carcinogenic for male or female B6C3F1 mice. Levels of Evidence of Carcinogenicity: Male Rats: Positive Female Rats: Positive Male Mice: Negative Female Mice: Negative Synonyms: cytembene; 2-butenoic acid; 3-bromo-4-(4 methoxyphenyl)-4-oxo-, sodium salt PMID- 12778229 TI - Carcinogenesis Bioassay of 1,2-Dibromo-3-chloropropane (CAS No. 96-12-8) in F344 Rats and B6C3F1 Mice (Inhalation Study). AB - 1,2-Dibromo-3-chloropropane (DBCP), a contaminant (0.05%) of the flame retardant tris(2,3-dibromopropyl)phosphate, has been used primarily as a soil fumigant to control nematodes. Unlike other halogenated nematocides, DBCP can be applied to soil without damaging growing perennials. Since it is slightly soluble in water at the concentrations used (30 ppm), DBCP can be either injected directly into the soil or added to irrigation water. By 1972, an estimated 12.3 million pounds were being used annually; in 1977, a total of 832,000 pounds were used in California, mostly on grapes and tomatoes. A carcinogenesis bioassay of technical grade 1,2-dibromo-3-chloropropane (DBCP), which contained trace amounts of epichlorohydrin and 1,2-dibromoethane, was conducted by exposing groups of 50 F344 rats and B6C3F1 mice of each sex by inhalation to concentrations of 0.6 or 3.0 ppm DBCP for 6 hours per day, 5 days per week, for 76 to 103 weeks. Untreated chamber controls consisted of 50 rats and 50 mice of each sex. Surviving high dose rats were killed at week 84. Surviving high-dose female mice and low-and high-dose male mice were killed at week 76. Low-dose rats and female mice were killed at week 104. Accelerated mortality occurred in the high-dose groups of both species. Early deaths of high-dose rats and mice were associated with respiratory tract tumors. Interference with breathing and metastasis to the brain were major contributing factors in these deaths. Among male mice, accelerated mortality occurred in low-dose and control groups as well as in the high-dose group. Urogenital infection appeared to be associated with these deaths. Carcinomas, squamous-cell carcinomas, and adenocarcinomas of the nasal cavity and squamous-cell papillomas of the tongue each occurred in high-dose male rats at incidences significantly higher than those in the corresponding controls. Adenocarcinomas, adenomas, adenomatous polyps, and squamous-cell papillomas of the nasal cavity and adenomatous polyps of the nasal turbinates occurred in low dose male rats with significantly increased incidences relative to controls. Carcinomas and adenocarcinomas of the nasal cavity, squamous-cell papillomas of the tongue, squamous-cell papillomas and carcinomas (combined) of the pharynx, and adenomas of the adrenal cortex each occurred in high-dose female rats at incidences significantly higher than those in the corresponding controls. Also, adenomas and squamous-cell papillomas of the nasal cavity, adenomas of the adrenal cortex, and fibroadenomas of the mammary gland were increased significantly in low-dose female rats when compared with controls. Adenocarcinomas of the nasal cavity in high-dose female mice, papillary carcinomas in low-dose female mice, and carcinomas, squamous cell carcinomas of the nasal cavity, and alveolar/bronchiolar adenomas or carcinomas of the lung in high-dose male and female mice occurred at incidences significantly higher than those in the corresponding controls. Exposure to DBCP vapor was also associated with toxic tubular nephropathy in rats and mice of either sex and with proliferative changes in the nasal mucosa, lung, and forestomach in mice. Under the conditions of this bioassay, DBCP was carcinogenic for male and female F344/N rats, including increased incidences of nasal cavity tumors and tumors of the tongue in both sexes, and cortical adenomas in the adrenal glands of females. DBCP was carcinogenic in male and female B6C3F1 mice, including increased incidences of nasal cavity tumors and lung tumors. Levels of Evidence of Carcinogenicity: Male Rats: Positive Female Rats: Positive Male Mice: Positive Female Mice: Positive Synonyms: DBCP; dibromochloropropane; Nemagon; Fumazone PMID- 12778230 TI - Bioassay of 4,4'-Oxydianiline for Possible Carcinogenicity (CAS No. 101-80-4). AB - 4,4'-Oxydianiline is used in the manufacture of high temperature resistant metal adhesives, molding and machine parts, and insulators. A bioassay of this chemical for possible carcinogenicity was conducted by feeding diets containing 200, 400, or 500 ppm of the test chemical to groups of 50 male or female F344 rats and 150, 300, or 800 ppm to groups of 50 male or female B6C3F1 mice for 104 weeks. Matched controls consisted of 50 untreated rats and 50 untreated mice of each sex. All surviving animals were killed at 104 to 105 weeks. A dose-related decrement in mean body weight gain was observed for all groups of dosed rats and mice. Survival was significantly shortened in the high-dose female rats and in the low- and mid-dose female mice. In male and female rats, hepatocellular carcinomas or neoplastic nodules occurred at incidences that were dose-related, and the incidences in all dosed groups (except low-dose females) were higher than those in the controls. The occurrence of follicular-cell adenomas or carcinomas of the thyroid was dose-related. Among groups of male and female rats, the incidences in the mid- and high-dose groups of either sex were significantly higher than those of the corresponding controls. In male and female mice, adenomas in the harderian glands occurred in all dosed groups at incidences that were significantly higher than the incidence in the matched controls. In low-dose male mice and in high dose female mice, hepatocellular adenomas or carcinomas occurred at incidences significantly higher than those in the matched controls. In female mice, follicular-cell adenomas in the thyroid occurred with a positive linear trend, and in a direct comparison the incidence in the high-dose group was also significantly higher than that in the controls. Tumors occurring among male mice at increased incidences which could not be statistically related to the chemical were adenomas in the pituitary and hemangiomas of the circulatory system. Under the conditions of this bioassay, 4,4'-oxydianiline was carcinogenic for male and female F344 rats, inducing hepatocellular carcinomas or neoplastic nodules and follicular-cell adenomas or carcinomas of the thyroid. 4,4'-Oxydianiline was also carcinogenic for male and female B6C3F1 mice, including adenomas in the harderian glands, hepatocellular adenomas or carcinomas in both sexes, and follicular-cell adenomas in the thyroid of females. Levels of Evidence of Carcinogenicity: Male Rats: Positive Female Rats: Positive Male Mice: Positive Female Mice: Positive PMID- 12778231 TI - Bioassay of Benzoin for Possible Carcinogenicity (CAS No.119-53-9). AB - A bioassay of benzoin for possible carcinogenicity was conducted by incorporating the test chemical in diets of F344 rats and B6C3F1 mice. Benzoin is used as a photopolymerization catalyst, chemical intermediate, and flavor ingredient. Groups of 50 male rats were fed diets containing 125 or 250 ppm benzoin for 104 weeks, and similar groups of female rats received feed containing 250 or 500 ppm. Groups of 50 mice of each sex were fed diets containing 2,500 or 5,000 ppm, benzoin for 104 weeks. Groups of 50 untreated rats and mice of each sex were used as matched controls. Rats and mice of either sex probably could have tolerated higher doses. An increased incidence of lymphomas or leukemia occurred in dosed male rats, but the observed dose-related trend was not statistically significant. Mean body weights and clinical signs of low-dose, high-dose, and control male and female rats and male mice were comparable throughout the study. After week 44, mean body weights of dosed female mice were slightly lower (10% or less) than those of the controls. The incidences of lymphomas that occurred in male mice varied with each dose but were not statistically significant when compared with those of matched controls. Lymphomas or leukemias occurred in low-dose female mice at an incidence that was significant when compared with the matched controls. However, because the incidence of lymphomas or leukemias in the high dose female mice was not significant, the occurrence of these tumors was not clearly related to administration of the test compounds. Under the conditions of this bioassay, benzoin was not carcinogenic for F344 rats or B6C3F1 mice. Levels of Evidence of Carcinogenicity: Male Rats: Negative Female Rats: Negative Male Mice: Negative Female Mice: Negative Synonym: 2-hydroxy-1,2-diphenylethanone PMID- 12778232 TI - The history of health care in the Kingdom of Saudi Arabia with emphasis on pediatric surgery. AB - The Kingdom of Saudi Arabia (KSA) is a large country with a population approaching 20 million people. It is a relatively young country, only united in 1932. The health services started with limited resources and very small clinics, and gradually reached highly sophisticated modern hospitals, medical centers and cities. The rapid growth of health services has also been accompanied by a well planned saudization process in manpower, to run such services. This was reflected by investing in scholarships, training courses and finally establishing the Saudi Council for Health Specialties, which provide structured training programs. Pediatric surgery is one of the youngest specialties to grow in KSA. It only received attention after 1980. However, in over 20 years, it has rapidly grown to become a well-recognized specialty with approximately 50 pediatric surgery specialists and consultants practicing in the field. Also lately, the sub specialty fellowship in pediatric surgery was approved as a 3-year training program following general surgery training. In summary, this review will highlight the rapid growth of health services in Saudi Arabia and the vision for the future to provide a modern medicine to parallel the country's growth and civilization. PMID- 12778233 TI - Retroperitoneal minimally invasive endoscopic adrenalectomy in children. AB - Conventional adrenalectomy for tumor excision is one of the challenging tasks in adults as well as children with mounted postoperative complications. Advancing laparoscopic techniques with the new era of diagnostic image developments aids this kind of procedure performed mainly in adults with only few reports in children. Here we present a review of this considerably new approach in children, adrenalectomy achieved through endoscopic retroperitoneal minimally invasive technique, discussing the indications, the advantages and disadvantages, and a comparison between this approach and other endoscopic techniques performed for this type of surgery. PMID- 12778234 TI - Laparoscopy and small bowel obstruction in children. AB - Three issues are studied: 1. What is well-known about post-operative small bowel obstructions. The cost of this pathology, the complications associated (blood loss, enterocutaneous fistulas, major bowel resections, abdominal wall damages, death), the distribution of adhesions in the abdominal cavity, and previous surgeries. 2. Small bowel obstructions happen after laparoscopy and the incidence is more than 1% of patients: herniation of the small bowel through a trocar site, herniation of the omentum through a trocar site, peritoneal defect, spillage of stones and stercolitis. Suggestions to avoid small bowel obstruction after laparoscopy include the use of smaller trocars whenever possible, repairing the fascia under direct vision, carefully desufflating the abdominal cavity, avoiding bleeding, spilled stones and staples and washing the abdominal cavity at the end of the surgical procedure. 3. The treatment of small obstruction is feasible by laparoscopy, reliability, and results are studied with a multicentric GECI (Groupe d'Etude en Coelioscopie Infantile) and literature series. We obtained good results for 2/3 children. PMID- 12778235 TI - Idiopathic intussusception in infancy and childhood. AB - OBJECTIVE: Idiopathic intussusception is an important cause of abdominal pain, bleeding per rectum and intestinal obstruction in infancy and childhood. This aim of this study was to undertake a retrospective review of all children who presented with idiopathic intussusception over a 17-year period. METHODS: The medical records of children who presented with idiopathic intussusception from January 1984 through December 2000 at King Fahad National Guard Hospital, Riyadh, Kingdom of Saudi Arabia were reviewed. The data obtained included age, sex, clinical presentation, diagnostic investigations, mode of treatment, length of hospital stay and results. RESULTS: Thirty-three children (21 male, 12 female) presented with 37 episodes of intussusception. Their mean age was 8.4 months (range 5 hours to 36 months). The clinical features included rectal bleeding (81%), vomiting (78%), abdominal colic/pain (65%) and abdominal mass (62%). All cases were ileocolic intussusception with no leading point. Barium enema was attempted in 36 cases with success in 20 (56%). Laparotomy was required in 16 cases, manual reduction being successful in 11 (30%) and 6 (16%) had bowel resection. At surgery, after attempted barium reduction, 9 (56%) cases had the intussusception already reduced to the cecum. Seventy percent of the cases presented within 24 hours of onset of symptoms. The 4 recurrences in 3 children had successful enema reduction. There was no mortality but 3 operative cases required late surgery for adhesive intestinal obstruction including one requiring bowel resection. CONCLUSION: Idiopathic intussusception commonly presents as an ileo-colic type but is uncommon in our institution. The clinical features are classical, rectal bleeding being the most common. The majority presented within 24 hours of onset of symptoms and barium enema reduction was successful in 20 out of 36 cases in which it was attempted. Since most intussusceptions were already in the cecum at surgery after failed enema reduction, a repeat or delayed enema reduction could be considered in stable cases. Recurrent intussusception occurred in 3 non-operated cases and adhesive intestinal obstruction in 3 laparotomy cases. PMID- 12778236 TI - Open laparoscopic access for primary trocar using modified Hasson's technique. AB - OBJECTIVE: To describe the safety and efficacy of open laparoscopic access for the primary trocar using modified Hasson's technique for laparoscopic surgery in children. METHODS: All 100 laparoscopic procedures performed at King Khalid University Hospital, Riyadh, Kingdom of Saudi Arabia between January 1999 and April 2001 using modified Hasson's technique were prospectively evaluated. They were aged from 3 months to 12 years. RESULTS: One hundred children who had open laparoscopic access during the study period had diagnosis of acute appendicitis (n=57), impalpable undescended testes (n=29), gallstones (n=5), varicocele (n=3) and others (n=6). Three children had minor operative complications (2 cases of pre-peritoneal placement of trocar, which were recognized immediately and the other had omental bleeding). Two children had post-operative complications related to primary access (one port infection and other port site hematoma). Access to the abdominal cavity was generally secured in 3-12 minutes (average 4+/ 2). Clinic follow-up ranged from 3-14 months. CONCLUSION: Open laparoscopic access using modified Hasson's technique was associated with no major or life threatening complications. Minor operative (3%) and post-operative (2%) complications occurred in the first 100 cases. Modified Hasson's technique for the primary trocar for accessing the abdominal cavity is a safe and effective method, and is recommended for all laparoscopic procedures in children. PMID- 12778237 TI - Treatment and postoperative complication of 420 patients with congenital megacolon. AB - OBJECTIVE: Hirschprung's disease is still one of the most important problems of pediatric surgery and its treatment has its own different complications and difficulties. Therefore, pediatric surgeons use different methods in each case. The objective of this study was to review the advantages and disadvantages of the Swenson's procedure over the past 20-years. METHODS: In this survey, we reported 420 patients who in a 20-year interval (from 1981-2000) with the net diagnosis of Hirschprung's disease, were admitted to the pediatric surgery ward of Taleghani Medical Center, Tehran, Iran and operated on by Swenson's procedure. This survey has been carried out in 2 parts with descriptional method using patients' files and analytical method by using patients' observation and questionnaires. In the first step, they were 420 patients and in second step 216 patients were studied and the results of the first 10 years (190 cases) and the results of the second 10 years (230 cases) were analyzed alone and compared with each. RESULTS: The results revealed wound infection in 17 cases in the first 10 years (14.2%), with18 cases in the second (7.9%) giving a mean of 10.7%. Anastomosis leakage was 6.8% in the first and 2.2% in the second 10 years with a mean of 4.2%. Septicemia was 3.6% in the first and 0.9% in the second period with a mean of 4%. Intestinal obstruction was 4.2% in the first and 2.1% in the second 10 years with a mean of 3%. There was no mortality during the surgery and for one month after. The results of late complications are: rectal anastomosis stricture, 13% in the first and 3% in the second 10 years with a mean of 7.6%; inguinal hernia of 1.6% in the first and 0.4% in the second period with a mean of 1% and finally, the death rate due to enterocolitis was 1.6% in the first and 0.9% in the second 10 years. Altogether, 87% of patients and their parents were satisfied with bowel habits after operation; 10% were relatively satisfied and 3% were not satisfied at all. Age distribution at the time of diagnosis: 50% of 0-1 months; 23% of 1-12 months and 27% were above one-year-old. Three hundred and eleven patients (74%) were male and 109 (26%) were female. The youngest patient was 3-days-old and the oldest was a 14-year and 3-month-old boy. The most common denerved part of the intestine was sigmoid colon with 54.4% prevalence. CONCLUSION: Our results show that Swenson's method is a successful procedure in the treatment of Hirschprung's disease. PMID- 12778303 TI - [The diagnostic challenges in fecal incontinence]. PMID- 12778304 TI - [Pudendal neuropathy: correlation with demographic data, severity index and pressoric parameters in patients with fecal incontinence]. AB - BACKGROUND: Fecal incontinence stands for inability in maintaining the control of defecation to a socially acceptable and adequate time and place, resulting in unwanted release of gas, liquid or solid stool. The diagnosis needs multiple exams. Anorectal manometry is mandatory for this study. The correlation between manometry with electophysiological studies and symptoms is not yet clear in the literature. AIMS: Correlate values of anal manometry, pudental nerve terminal motor latency and co morbidity in fecal incontinence patients. METHODOLOGY AND PATIENTS: Patients with clinical fecal incontinence, who attended the outpatient department of "Hospital Nossa Senhora da Concei o", Porto Alegre, RS, Brazil, between March 1997 and June 2000, were studied prospectively. Every single patient has undergone a general investigation, incontinence score, anal manometry, bilateral pudendal nerve terminal motor latency and physical examination. For statistical purposes the patients were separated in groups according to the results of the anal manometry, pudendal neuropathy, for age and sex. RESULTS/CONCLUSION: Thirty-nine patients were studied, 85,6% female, average age 60,1 years (+/- 12,89). The average index of the fecal incontinence was of 9,30 (+/- 4,93). In the anorectal manometry 23 patients showed low pressure. The pressures were significantly higher among males. The nerve terminal motor latency and pudental nerve (neuropathy) time was greater in 14 patients (35,9%). The age and the time of pudental nerve terminal motor were significantly co related. The other co relations were not statistically significant. PMID- 12778305 TI - [Outcome of liver transplantation in patients with alcoholic liver disease]. AB - BACKGROUND: Liver transplantation is accepted as effective therapeutic option for end-stage liver disease, including alcoholic liver disease AIM: To evaluate the outcome of liver transplantation for alcoholic liver disease in the Liver Transplantation Program at "Hospital de Clinicas" of the Federal University of Parana, Curitiba, PR, Brazil. PATIENTS AND METHODS: It was performed a retrospective study of the patients who underwent liver transplantation for alcoholic end-stage liver disease between September 1991 and January 2001. The minimum abstinence period required was 6 months before liver transplantation. Identification of alcohol consumption after liver transplantation was determinated by information provided by patient or family and biochemical or histological anormalities. RESULTS: Twenty patients underwent liver transplantation for alcoholic liver disease in the study period, 95% (19/20) were men and the median age was 50 years (29-61 years). Seventy-five percent of the patients (15/20) had severe liver disfunction (Child C class) in the pre transplant period. In six of them (30%) there was association with viral hepatitis and in one, with hepatocarcinoma. Median abstinence period before liver transplantation was 24 months, varying from 9 to 120 months. One-year and 3-year survival rate were 75% and 50%, respectively. The main complications were: acute cellular rejection (40%), chronic rejection (5%), hepatic artery thrombosis (15%), biliary complications (15%), bacterial or fungal infections (45%), cytomegalovirus infection (20%). Three patients returned to alcohol use after liver transplantation CONCLUSION: The survival of patients who received liver transplant for alcoholic cirrhosis are satisfactory. In the present study there was a small index of alcohol use after liver transplantation. PMID- 12778306 TI - [Efficacy of contrast chromoendoscopy of the colon after oral administration of indigo carmine dye]. AB - INTRODUCTION: Indigo carmine dye is usually spread directly over the colon in many chromoscopic techniques aiming better visualization of a lesion already detected by conventional colonoscopy. Examination of the colon already stained by oral administration of indigo carmine dye may increase detection of small lesions resulting in higher sensibility of the colonoscopy in diagnosing diminutive lesions. OBJECTIVE: Analyze the results regarding the quality of chromoscopic technique and the indigo carmine dye distribution over the colon after oral administration. PATIENTS AND METHODS: Fifty patients undergoing colonoscopy were evaluated. A capsule containing 100 mg of indigo carmine dye was offered to these patients 30 min before oral mannitol prep routinely used. The indigo carmine dye contrast effect was graded as bad, regular or good according to preestablished criteria in three segments of the colon: right and left colon and the rectum. RESULTS: In the right colon, good indigo carmine dye contrast effect was observed in only 9 (18.8%) patients, while it was considered regular and bad in 32 (66.6%) and in 7 (14.6%) patients, respectively. A good indigo carmine dye contrast effect was never observed in this series for the left colon or in the rectum. As a matter of fact, no indigo carmine dye was observed in the left colon in 80.9% and in the rectum in 92% of patients in this series. CONCLUSION: Although it may be simple and desirable, oral administration of indigo carmine dye seems ineffective for enhancing detection of diminutive lesions by chromoscopy as result of poor colonic distribution of indigo carmine dye mainly at distal colonic sites. PMID- 12778307 TI - [Prevalence and prognosis of spontaneous bacterial peritonitis. Experience in patients from a general hospital in Porto Alegre, RS, Brazil (1991-2000)]. AB - BACKGROUND: Spontaneous bacterial peritonitis is a frequent complication that occurs in patients with cirrhosis and ascites and has a recurrence rate of 70% in 1 year. In addition, this infection determines a poor short and long-term prognosis and a shorter survival rate. AIMS: Evaluate the prevalence of spontaneous bacterial peritonitis in cirrhotic patients with ascites and the effect of its occurrence on the survival. PATIENTS/METHODS: One thousand and thirty admissions of patients with cirrhosis and ascites were reviewed and 114 episodes of spontaneous bacterial peritonitis were documented in 94 patients. The ascitic analysis was accomplished in all patients. The diagnosis of this infection was established when the ascitic fluid polymorphonuclear count was equal or above 250 cells mm3. RESULTS: The prevalence of this infection was 11.1% and the mortality rate 21.9%. Spontaneous bacterial peritonitis was community acquired in 61.4% and hospital acquired in 37.7%. The mortality rate was 18.6% and 27.9%, respectively. The infection resolved in 91.1% of the episodes by the analysis of ascitic fluid at 48 hours on antibiotics. The use of prophylactic antibiotics was documented in 22.3% of the episodes, but there are not significant differences on the mortality or type of bacteria isolated when comparing the patients with or without this treatment. CONCLUSIONS: Spontaneous bacterial peritonitis is a common complication in patients with cirrhosis and ascites and determines a worse prognosis, mainly when related with absence of initial response to antibiotics. PMID- 12778308 TI - [Clinical and macroscopic variables that influence the prognosis of colorectal carcinoma]. AB - BACKGROUND AND AIMS: The paradoxical evolution of approximately one third of patients with neoplasms cataloged in Dukes stages B and C demonstrates the desirability of utilizing other prognostic criteria that are capable of broadening the information provided by these two important variables. Only a small number of investigators have dedicated themselves to the study of the prognostic value of clinical and macroscopic parameters of colorectal neoplasms, and the results obtained have been shown to be controversial. The principal aim of this work was to evaluate the prognostic importance of these parameters. PATIENTS AND METHODS: A study was made of 320 patients with colorectal cancer who underwent curative extirpation. They had a median age of 58 years, and there were 199 females (62.2%) and 121 males (37.8%). The patients were divided into three age groups: under 40 years old, between 40 and 60 years old and over 60 years old. The tumors were distributed in three intestinal segments: right colon, left colon and rectum. The neoplasms were classified as small (diameter less than or equal to 35 mm) and large (diameter greater than 35 mm). With regard to their form, they were classified as exophytic, when characterized by luminal growth, and endophytic, when there was intramural growth. The involvement of the intestinal circumference at the site of the neoplasm was considered as partial or total. RESULTS: Of the 320 patients, 22 (6.9%) were aged under 40 years, 159 (49.7%) from 40 to 60 years and 139 (43.4%) presented an age of over 60 years. Seventy-three (22.8%) of the neoplasms were located in the right colon, 130 (40.6%) in the left colon and 117 (36.6%) in the rectum. Regarding the size, 280 (87.5%) were large and 40 (12.5%) small; exophytic lesions predominated over endophytic ones - 173 (54.1%) vs 147 (45.9%). A greater number of tumors presented total involvement of the intestinal circumference - 216 (67.5%) - while 104 (32.5%) presented partial involvement. The 5-year survival of the patients was not influenced by their age and sex, or by the location and size of the neoplasms. Exophytic lesions conferred greater survival on their sufferers (65.9%), in comparison with endophytic lesions (49.0%). The survival of patients with lesions partially involving the intestinal circumference was greater than for those with total involvement - 72.1% vs. 51.4%. CONCLUSIONS: Clinical variables had no influence on the patients' prognosis. Among the macroscopic variables, the form of the neoplasia and its involvement in the intestinal circumference did influence the patients' prognosis. These last two variables are important data capable of contributing to the identification of patient subpopulations with greater or lesser prognostic risk. PMID- 12778309 TI - [Ulcerative esophagitis associated with the use of alendronate sodium: histopathological and endoscopic features]. AB - BACKGROUND: Drug-induced or "pill-induced" esophagitis may be secondary to the prolonged contact of the drug with the esophageal mucosa or secondary to the drug ability to alter the local conditions. The alendronate sodium, a bone resorption inhibitor used in the treatment and prevention of osteoporosis, has been cited, recently, as one of the causes of adverse upper gastrointestinal tract injury. AIM: To describe the clinical, endoscopic and histopathological features of patients with ulcerative esophagitis associated with alendronate sodium. PATIENTS: Four women and one man with osteoporosis were treated with alendronate sodium and submitted to endoscopy followed esophageal biopsy. RESULTS: The age range of the patients was from 64 to 84 years old. The patients showed dyspeptic symptoms after taking alendronate sodium during a period of 2-12 months. At endoscopic evaluation, the mucosa was friable, with erosion and/or ulceration covered by fibrin in the distal esophagus. The pathological examination of the esophageal biopsies revealed ulcerative esophagitis characterized by necrofibrinpurulent material, granulation tissue, and yellow refractile polarizable crystal. The patients' symptoms resolved after stopping alendronate sodium use. CONCLUSIONS: The esophagus injuries associated with alendronate sodium are not frequent and seem to be associated with the incorrect use of medication. The endoscopists and pathologists should be alert to the possibility of alendronate sodium therapy in cases of diagnosis of ulcerative esophagitis in ancient patients, particularly in women. The recognition of this condition would improve the patient care. PMID- 12778310 TI - Infectious endocarditis from Streptococcus bovis associated with colonic carcinoma: case report and literature review. AB - BACKGROUND: Many studies in the literature have warned of the need for investigation of colonic lesions among patients, especially elderly ones, who have bacteremia and/or endocarditis from Streptococcus bovis. Bacteremia and infectious endocarditis from Streptococcus bovis may be related to the presence of neoplastic lesions in the large intestine and hepatic disease. AIM: This report describes a patient who presented infectious endocarditis from Streptococcus bovis associated with colonic carcinoma and tubular-villous adenomas. CONCLUSIONS: The finding of this bacterium among patients with septicemia and/or endocarditis is also related to the presence of villous or tubular-villous adenomas in the large intestine. For this reason, complete and detailed investigation of the large intestine must be performed in patients with infectious endocarditis, even in the absence of intestinal symptoms. An increased incidence of this condition or hepatic dysfunction has been reported among patients with infectious endocarditis from Streptococcus bovis. Patients with infectious endocarditis from Streptococcus bovis and normal colonoscopy may be included in the group at risk for developing colonic cancer. The knowledge that there is an association between endocarditis from Streptococcus bovis and carcinoma of the colon has important clinical implications. If the lesion can be discovered at an early stage, curative resection may become possible. PMID- 12778311 TI - [Subjective nutrition assessment: Part 1 - A review of its validity after two decades of use]. AB - BACKGROUND: The subjective global assessment is a clinical method for nutritional assessment that considers not only body composition alterations but also changes in physiological function. The method is simple, inexpensive and non-invasive, and it can be performed at bedside, and its use was described for the first time almost two decades ago. AIM: To review the validity studies of subjective global assessment described in literature in the last two decades of its use. METHODS: It was performed a systematic review in MEDLINE, using "subjective global assessment" as search term and the most relevant papers were selected. RESULTS: Being a subjective method, its precision depends on the observer's experience. Nevertheless, the method showed a good diagnostic precision when performed by trained observers. Subjective global assessment was validated by convergent validity, when this method was compared to other objective nutritional assessment methods, and by predictive validity, showing that subjective global assessment could identify patients who were at high risk for developing postoperative complications. CONCLUSIONS: Subjective global assessment has been a good option in nutritional assessment in surgical patients and some modifications have been suggested to adjust the method to other clinical situations. The observer experience is of extreme importance, since the precision of the method depends on it. PMID- 12778312 TI - [Damage control: a tactical alternative for the management of exanguinating trauma patients]. AB - BACKGROUND: Despite the advances in the treatment of exanguinating patients, hemorrhage remains as the leading cause of early deaths. A great deal of attention has been given to "damage control" as a therapeutic alternative in this scenario. AIM: To appraise the definition, indications, operative techniques and results of damage control for the treatment of exanguinating trauma patients. METHOD: Bibliographic review. RESULTS: Damage control introduces the concept of breaking the vicious cycle of metabolic acidosis, hypothermia and coagulopathy which results from hemorrhagic shock. Thus, the operation has to be interrupted before this irreversible stage, even if the injured organs were not given the definitive treatment at this moment. So, damage control involves three steps: an abbreviated operation, a recovering period in the intensive care unit, and the reoperation for the definitive treatment. At the abbreviated operation, operative techniques as stapling intestinal injuries or packing liver wounds are applied, allowing rapid control of the bleeding and spillage. In the intensive care unit, the patient is warmed, oxygen delivery and consumption are restored and coagulation factors administered. As soon as the hemodynamic stability, ideal body temperature and coagulation status are reached, the definitive operation is carried out. Damage control is a helpful option if correctly used. However, there are also severe complications that can occur. Therefore, it should be employed only in centers that could provide optimum resources. CONCLUSION: Damage control is an important tactical alternative for the treatment of exanguinating trauma patients. PMID- 12778313 TI - [Evaluation of enzyme-linked immunosorbent assay for the diagnosis of Helicobacter pylori infection in children and adolescents]. AB - BACKGROUND: Helicobacter pylori infection is recognized as the most frequent cause of chronic gastritis in adults and children. The diagnosis is accomplished with invasive methods in fragments of endoscopic gastric biopsies and non invasive methods. The enzyme-linked immunosorbent assay constitutes a simple, fast exam and of low cost with high sensibility in adult patients. AIM: The purpose of this study was to evaluate the ELISA method for the infection diagnosis for Helicobacter pylori in children and adolescents using the Cobas Core II kit (Roche). Helicobacter pylori was positive when the rapid urease test and the histology were both positive or when the culture was positive and Helicobacter pylori negative when all the tests were negative. PATIENTS/METHODS: Eleven hundred patients were studied, their age ranged from 3 months and 16 years, (mean = 8y 7m +/-; 4.0; median = 9y 5m). Helicobacter pylori infection was diagnosed in 47.7% (53/111). RESULT: Sensitivity was 83.0% and 86.0%; specificity was 70.6% and 71.0%, using the cutoff of 7 U/mL and 5 U/mL, respectively. When only the positive culture was used as gold standard and the cutoff of 5 U/mL, the sensitivity was 93.3%. In patients older than 10 years, the sensibility was 90.6% and 96.8%; specificity was 71.0% and 61.9%, with the cutoff of 7 U/mL and 5 U/mL, respectively. CONCLUSION: ELISA method had good sensitivity in children older than 10 years, using the cutoff 5 U/mL, but the specificity was low. PMID- 12778314 TI - [Inguinal hernia repair in children: importance of combined local anesthesia]. AB - AIM: To describe an anesthetic technique, as well as the results of surgical treatment of the inguinal hernia in children. PATIENTS/METHODS: Forty-eight patients were submitted to inguinal hernia repair under local anesthesia at "Santa Casa de Misericordia de Cerqueira Cesar", State of Sao Paulo, Brazil. There were 34 male and 14 female patients, range from 3 months to 12 years old. Local anesthesia was performed with a dose of 5 mg/kg body weight of 1% lidocaine through iliohypogastric and ilioinguinal nerve blocks, medially to the anterior superior iliac spine, and at level of the pubic tubercle. Sedation was done with an association of ketamine (1 to 2 mg/kg) and diazepam (0,2 to 0,4 mg/kg). RESULTS: In all patients except one the procedure was done without complications. In that patient the local anesthesia was not effective and was followed by inhalatory anesthesia. Surgical complications (blood collections) were observed in three patients: two at the scrotum and one at subcutaneous, with good evolution. CONCLUSIONS: We conclude that the use of local anesthesia associated to sedative is a simple and safe procedure for the inguinal hernia repair in children. PMID- 12778315 TI - Expression of a novel bHLH-Zip gene in human testis. AB - AIM: To identify specifically expressed genes in the adult and fetal testes. METHODS: A human testis cDNA microarray was established. Then the mRNA of adult and fetal testis was purified and probes were prepared by a reverse transcription reaction with the testis mRNA as template. The microarray was hybridized with probes of adult and fetal testes. The nucleic sequences of differentially expressed genes were determined and homologies were searched in the databases of the GenBank. RESULTS: When hybridized with adult or fetal testis probes, the positive clones were 96.8 % and 95.4 %, respectively. Among these genes, one was a new testis-specific gene, which was named TSP1. TSP1 was highly expressed in human adult testis. The cDNA of TSP1 was 1,484 bp in length. The cDNA sequence of this clone was deposited in the Genbank (AF333098). TSP1 was also determined as Interim Gen Symbol (Unigene, No. Hs.98266). Protein analysis showed that TSP1 contained two functional domains: an N-terminal basic helix-loop-helix (bHLH) and a C-terminal leucine zipper (Zip). Homologous analysis showed that the 430 amino acid sequences deduced from the 1293 bp open reading frame (ORF) had a homology with the human gene FLJ2509 (AK098575). TSP1 had also a sequence homology with Spz 1 protein of mouse. Expression profiles showed that TSP1 was specifically and strongly expressed in the testis. CONCLUSION: TSP1 is a gene highly expressed in adult testis. It may play an important role in spermatogenesis in the humans. PMID- 12778316 TI - Effect of age on quality of fresh and frozen-thawed semen in White Italian ganders. AB - AIM: To comparatively evaluate the fresh semen quality of 1, 2 and 3-yr-old White Italian ganders (Anser anser L.) and the susceptibility of spermatozoa to freezing-thawing procedure. METHODS: Semen was collected by dorso-abdominal massage every 2 days~3 days from three groups of ganders: 1-yr-old (n=11), 2-yr old (n=7) and 3-yr-old (n=9). In the pooled fresh semen samples, the following parameters were evaluated: the ejaculate volume, the blood or feca contamination and the motility, concentration and morphology of spermatozoa. Sperm motility and morphology were evaluated in the frozen-thawed semen. Semen diluted with EK extender was frozen in straws in a computerized freezing unit with 6 % dimethyl formamide to -140 deg at a rate 60 deg/min and then transferred into the LN2 container. Straws with semen were thawed in a water bath at 60 deg. RESULTS: The ejaculate volume decreased with the age (0.21 mL for 1-yr-old, 0.18 mL for 2-yr old and 0.14 mL for 3-yr-old ganders); the sperm concentration increased with the age (327 x 10(6) mL(-1) for 1-yr-old, 431 x 10(6) mL(-1) for 2-yr-old and 547x10(6) mL(-1) for 3-yr-old ganders); the number of live - normal sperm was significantly (P<0.01) lower in the 1-yr-old than that in the 2- and 3-yr-old ganders (26.61 %, 41.54 and 35.9 %, respectively). The percentage of normal cells survived the freezing-thawing process was 37.7 %, 43.3 % and 40.9 % for 1-, 2- and 3-yr-old ganders, respectively. CONCLUSION: Freezing and thawing processes more significantly (P<0.01) affected the motility, viability and morphology of spermatozoa in semen of 1-yr-old ganders in comparison with older males. PMID- 12778317 TI - Potential role of reactive oxygen species on testicular pathology associated with infertility. AB - AIM: To investigate the level of malondialdehyde (MDA), a direct indicator of lipid peroxidation-induced injury by reactive oxygen species (ROS), in testicular biopsy specimens from infertile patients. METHODS: Levels of MDA were measured in testicular biopsy specimens from 29 consequent-randomized infertile men, aged 29.58+/-4.76 (21 to approximately 45) years. All patients were evaluated by a complete medical and reproductive history, physical examination, semen analysis (at least two), serum follicle-stimulating hormone and free testosterone levels, testicular biopsy and contact imprint. Scrotal colour Doppler ultrasonography was used to confirm suspected varicocele. The testicular MDA level was measured using the thiobarbituric acid test and the results were expressed per unit tissue weight. RESULTS: As a causal factor in infertility, varicocele was identified in 17 (58.6 %) patients, and idiopathic infertility, testicular failure and obstruction in 4 (13.8 %) patients each. The testicular MDA level was 13.56 (6.01), 49.56 (24.04), 58.53 (48.07), and 32.64 (21.51), 32.72 (13.61), 23.07 (7.82), 42,12 (34.76) pmol/mg tissue in the normal spermatogenesis (control), late maturation arrest, Sertoli cell only (SCO) and hypospermatogenesis (mild, moderate, severe) groups, respectively. The elevation of MDA levels was significant in the testicular tissue from SCO and maturation arrest groups compared with the controls (P<0.05). In addition, the elevation in testicular MDA levels between the SCO and the moderate hypospermatogenesis, and the moderate hypospermatogenesis and the maturation arrest groups was significant (P<0.05). CONCLUSION: Severe pathologic changes in the testicular tissue are associated with a high level of lipid peroxidation. These findings suggest that overproduction of ROS may play a role in the mechanism of testicular degeneration associated with infertility. PMID- 12778318 TI - Seminal concentrations of trace elements in various animals and their correlations. AB - AIM: To determine the seminal concentrations of copper, zinc, iron, cadmium, lead and nickel in bulls, rams, boars, stallions and foxes and study their correlations. METHODS: Semen samples were obtained, digested and analyzed by means of the atomic absorption spectrophotometer. Data were analyzed statistically with the Student's t-test and Scheffe's test using PC programs SAS and Excel. RESULTS: The seminal copper concentration was significantly higher in ram [(2.49+/-0.18) mg/kg] and fox [(2.16+/-0.53) mg/kg] than that in bull [(1.64+/-0.21) mg/kg], boar [(1.64+/-0.28) mg/kg] and stallion (0.86 mg/kg). In boar a significantly higher seminal zinc concentration (171.74+/-65.72) mg/kg] was found in comparison with stallion [(86.20+/-45.88) mg/kg], bull [(83.15+/ 61.61) mg/kg], ram [(60.46+/-35.37) mg/kg] and fox (13.09+/-5.22) mg/kg]. The iron concentration in semen was significantly higher in ram [(40.32+/-10.81) mg/kg), bull [(38.04+/-22.07) mg/kg] and fox [(33.16+/-24.36) mg/kg] than that in boar [(16.14+/-10.35) mg/kg] and stallion (12.68 mg/kg). The seminal cadmium concentration was relatively low [(0.05~0.12) mg/kg] in all studied species. The seminal lead concentration was the highest in ram [(0.35+/-0.68) mg/kg], which was much higher than in fox [(0.08+/-0.06) mg/kg], bull [(0.06+/-0.04) mg/kg], stallion [(0.05+/-0.05) mg/kg] and boar [0.02+/-0.03) mg/kg]. The level of seminal nickel was significantly higher in fox [(0.35+/-0.24) mg/kg] and ram [(0.31+/-0.19) mg/kg] in comparison with bull [(0.12+/-0.07) mg/kg] and boar [(0.06+/-0.08) mg/kg]. The concentration of nickel in the semen of stallion was (0.20 +/-0.24) mg/kg. There was a high positive correlation between seminal iron and zinc in bull (r=0.723) and stallion (r=0.723), between cadmium and lead in ram (r=0.976) and boar (r=0.973) and between iron and cadmium (r=0.783) and iron and lead (r=0.791) in boar. A high negative correlation between seminal nickel and copper in ram (r=-0.709) and between seminal copper and lead in fox (r= 0.854) was found. CONCLUSION: There are significant differences in the concentrations of seminal elements in different animals. There is a high concentration of seminal copper in ram and fox, a high zinc level in boar, a high iron level in bull, ram and fox and a high nickel level in ram and fox. PMID- 12778319 TI - Effect of a single dose of malathion on spermatogenesis in mice. AB - AIM: To observe the acute effect of the organophosphorous insecticide malathion on testicular function in mice. METHODS: The effects of a single dose of malathion [240 mg/kg (1/12 LD(50))] on plasma acetylcholinesterase (ACE) activity, spermatozoa (epididymal cauda counts and teratozoospermia), testis and plasma testosterone concentration) were evaluated at day 1, 8, 16, 35 and 40 after treatment. RESULTS: The sperm count was decreased significantly 24 h after treatment and teratozoospermia was increased at day 35 and 40. The height of the seminiferous epithelium and the diameter of tubular lumen were decreased at day 8. The percentage of tubular blockade was increased between day 8 and 35. A decrease in testosterone plasma level was observed at day 16 after treatment. CONCLUSION: Malathion damages male reproduction. The depletion of seminiferous tubules and the increase in teratozoospermia may be a genotoxic damage to the renewing spermatogonia, but the possibility of spermatogenic/spermiogenic disfunction due to a decrease in the plasma testosterone level can not be ruled out. PMID- 12778320 TI - Effect of experimental varicocele on structure and function of epididymis in adolescent rats. AB - AIM: To study the effect of experimental left varicocele (ELV) on epididymal structure and function in adolescent Sprague-Dawley rats. METHODS: ELV was induced by partial ligation of the left renal vein. Sham-operated animals served as the controls. Four and 8 weeks after the operation, the histological, ultrastructural and biochemical (alpha-glucosidase activity and carnitine content) changes in different segments of the epididymis were observed. RESULTS: In the treated animals, there were degeneration of the epididymal epithelium and edema of the interstitial tissue; numerous shedding cells, residual bodies, deformed sperm and macrophages appeared in the epididymal lumen. Morphometric measurement indicated a significant reduction in the epididymal tubular diameter (P<0.05) and a significant increase in the epididymal interstitial area (P<0.05) compared with the controls. Ultrastructural study showed sparse microvilli of the columnar epithelium, increased and enlarged lysosomes in the principal cells with defected organelles and the presence of large cytoplasmic vacuoles. The protein and carnitine contents and the alpha-glucosidase activity in the caput, corpus and cauda epididymis of the ELV rats were lower than those of the controls (P<0.05). CONCLUSION: There were structural and functional changes in the epididymis of adolescent ELV rats, which may contribute to the infertility caused by varicocele. PMID- 12778321 TI - Influences of dibutyryl cyclic adenosine monophosphate and forskolin on human sperm motility in vitro. AB - AIM: To study the influences of dibutyryl cyclic adenosine monophosphate (dbcAMP) and forskolin on human sperm motility in vitro. METHODS: Semen samples, aseptically obtained by masturbation and prepared by swim-up technique from 20 fertile men, were incubated with different concentrations of dbcAMP and forskolin at 37 deg. Measurements were carried out after 10 min, 20 min, 30 min and 60 min incubation. Motility parameters were estimated by using an automatic analyzing system. RESULTS: Treatment with dbcAMP or forskolin resulted in a significant increase in sperm motility and progressive motility. The larger the concentrations of dbcAMP or forskolin, the greater the effect appeared. The straight linear velocity and curvilinear velocity were not affected by both agents. CONCLUSION: dbcAMP and forskolin increase the motility and progressive motility of human sperm in vitro. PMID- 12778322 TI - Effect of aging on expression of nitric oxide synthase I and activity of nitric oxide synthase in rat penis. AB - AIM: To investigate the effect of aging on the expression of nitric oxide synthase I (NOS I) and the activity of NOS in rat penis. METHODS: Sixty male rats from 3 age groups (adult, old and senescent) were investigated. The expression of NOS I protein and mRNA in rat penis were detected by Western blot and RT-PCR respectively and the NOS activity, with ultraviolet spectrophotometry. RESULTS: In the old and senescent group, NOS I protein expression was significantly decreased as compared with the adult. NOS I mRNA expression was well correlated with the protein expression. NOS activity was not statistically different between the adult and old groups, but it was significantly reduced in the senescent compared with the adult group (P<0.01). CONCLUSION: The aging-induced decreases in NOS I expression and NOS activity may be one of the main mechanisms leading to erectile dysfunction in the senescent rats. PMID- 12778323 TI - Effect of Semecarpus anacardium fruits on reproductive function of male albino rats. AB - AIM: To evaluate the effect of an ethanolic extract of Semecarpus anacardium fruits on spermatogenesis in albino rats. METHODS: Male albino rats were fed with a 50 % ethanolic extract of Semecarpus anacardium fruit at 100 mg.kg(-1).day(-1), 200 mg.kg(-1).day(-1) and 300 mg.kg(-1).day(-1) for 60 days. Fertility test was performed after 60 days of treatment. Sperm motility and density were observed in the cauda epididymis. Biochemical and histological analyses of the blood and reproductive organs were done. Recovery of fertility was followed to evaluate the reversibility of drug action. RESULTS: S. anacardium fruit extract administration resulted in spermatogenic arrest in albino rats. The sperm motility and density was reduced significantly. The RBC and WBC counts, haemoglobin, haematocrit, blood sugar and urea were found to be within the normal range in the whole blood. The protein, cholesterol and glycogen in the testes and the fructose in the seminal vesicle were significantly decreased after the treatment. The fruit extract feeding caused marked reduction in the number of primary spermatocytes, secondary spermatocytes and spermatids. The number of mature Leydig cells was also decreased and degenerating cells increased proportionately. CONCLUSION: S. anacardium fruit extract causes spermatogenic arrest in albino rats. PMID- 12778324 TI - Effect of Stephania hernandifolia leaf extract on testicular activity in rats. AB - AIM: The testicular inhibitory effect of the aqueous fraction of methanol extract of Stephania hernandifolia leaf was studied in male Wistar rats. METHODS: The supernatent and the precipitate part of aqueous fractions of the methanol extract of the leaf were gavaged separately to rat at a similar dose of 200 mg/mL per 100 g body weight per day for 28 days. After cessation of treatment, various observations were conducted. RESULTS: In both treated groups, there were significant decreases in the relative weights of the sex organs, the testicular key androgenic enzymes activities, the plasma level of testosterone, the number of different germ cells at stage VII of seminiferous epithelial cell cycle and the seminiferous tubular diameter in comparison to the controls. Neither of the parts had somatic, renal and hepatic toxicity. This study suggested that the active molecules present in the aqueous fraction of methanol extract of Stephania hernandifolia leaves might be steroids as indicated by thin layer chromatography using specific staining substance for steroid molecules. CONCLUSION: In rats, the aqueous fraction of methanol extract of the S. hernandifolia leaves possesses certain testis-inhibitory substances, which may be steroid-like agents. PMID- 12778325 TI - Sperm immobilization activity of Allium sativum L. and other plant extracts. AB - AIM: To identify possible spermicidal agents through screening a number of edible medicinal plants with antimicrobial activity. METHODS: Initial screening was made on the basis of ram cauda epididymal sperm immobilization immediately after addition of extracts. The most potent extract was selected and was evaluated on both ram and human spermatozoa. To unravel its mode of action several sperm functional tests were carried out, namely viability of cells, hypo-osmotic swelling test for membrane integrity and assays of membrane-bound enzyme 5' nucleotidase and acrosomal marker enzyme acrosin. RESULTS: The crude aqueous extract of the bulb of Allium sativum L. showed the most promising results by instant immobilization of the ram epididymal sperm at 0.25 g/mL and human ejaculated sperm at 0.5 g/mL. Sperm immobilizing effects were irreversible and the factor of the extract responsible for immobilization was thermostable up to 90 deg. On boiling at 100 deg for 10 minutes, this activity was markedly reduced. Moreover, this extract was able to cause aggregation of ram sperms into small clusters after 30 minutes of incubation at 37 deg. However this property was not found in human spermatozoa. More than 50 % reduction in sperm viability and hypo osmotic swelling occurred in treated sperm as compared with the controls, indicating the possibility of plasma membrane disintegration which was further supported by the significant reduction in the activity of membrane bound 5' nucleotidase and acrosomal acrosin. CONCLUSION: The crude aqueous extract of A. sativum bulb possesses spermicidal activity in vitro. PMID- 12778326 TI - Searching for candidate genes for male infertility. AB - AIM: We describe an approach to search for candidate genes for male infertility using the two human genome databases: the public University of California at Santa Cruz (UCSC) and private Celera databases which list known and predicted gene sequences and provide related information such as gene function, tissue expression, known mutations and single nucleotide polymorphisms (SNPs). METHODS AND RESULTS: To demonstrate this in silico research, the following male infertility candidate genes were selected: (1) human BOULE, mutations of which may lead to germ cell arrest at the primary spermatocyte stage, (2) mutations of casein kinase 2 alpha genes which may cause globozoospermia, (3) DMR-N9 which is possibly involved in the spermatogenic defect of myotonic dystrophy and (4) several testes expressed genes at or near the breakpoints of a balanced translocation associated with hypospermatogenesis. We indicate how information derived from the human genome databases can be used to confirm these candidate genes may be pathogenic by studying RNA expression in tissue arrays using in situ hybridization and gene sequencing. CONCLUSION: The paper explains the new approach to discovering genetic causes of male infertility using information about the human genome. PMID- 12778327 TI - Androgen and bone mass in men. AB - Androgens have multiple actions on the skeleton throughout life. Androgens promote skeletal growth and accumulation of minerals during puberty and adolescence and stimulate osteoblast but suppress osteoclast function, activity and lifespan through complex mechanisms. Also androgens increase periosteal bone apposition, resulting in larger bone size and thicker cortical bone in men. There is convincing evidence to show that aromatization to estrogens was an important pathway for mediating the action of testosterone on bone physiology. Estrogen is probably the dominant sex steroid regulating bone resorption in men, but both testosterone and estrogen are important in maintaining bone formation. PMID- 12778328 TI - Biofeedback therapy for chronic pelvic pain syndrome. AB - AIM: To evaluate the efficacy of biofeedback therapy in patients with chronic pelvic pain syndrome (CPPS). METHODS: From November 2001 to April 2002, patients visiting the Urological Outpatient Clinic of this Hospital were evaluated by means of the National Institute of Health Chronic Prostatitis Symptom Index (NIH CPSI) and classified by the NIH classification standard. Sixty-two patients of CPPS category III were involved in this study. All patients had been treated by conventional approaches such as antibiotics and alpha-blockers for more than half a year without any improvement. The expressed prostatic secretion results were as follows: WBC 5 to 9/high power field, lipid + approximately +++ and bacterial culture negative. Their NIH-CPSI were 12 approximately 40. All the 62 cases complained of micturitional irritation (frequency, urgency, splitted stream and sense of residual urine), 32 cases, of pain or discomfort at the testicular, penile, scrotal, pelvic or rectal region and 13 cases, of white secretion dripping. The patients were treated by the Urostym Biofeedback equipment (Laborie Co., Canada) 5 times a week for 2 weeks with a stimulus intensity of 15 mA approximatley 23 mA and duration of 20 minutes. RESULTS: Sixty patients were significantly improved or cured, while no significant improvement in the remaining 2. No apparent side effect was observed. The NIH-CPSI dropped to 6 to 14 with an average reduction of 21 (P<0.01). In the 60 improved cases, pain was relieved after 2 approximately 3 treatment courses and other symptoms disappeared after 4 approximately 5 courses. CONCLUSION: Biofeedback therapy is a safe and effective treatment for CPPS. Large randomized clinical trials are needed to confirm its efficacy and to explore the mechanism of action. PMID- 12778329 TI - Folding and everting distal end of graft flap to reduce orifice stenosis following onlay urethroplasty. AB - AIM: Orifice stenosis remained to be a common complication of hypospadias repair. We had modified the preputial island flap urethroplasty by folding and everting the distal end of the pedicle graft flap to prevent the neo-orifice from stenosis. METHODS: Sixteen patients had undergone hypospadias repair using a modified onlay island flap technique. A urethral catheter was retained for 8 days to 10 days after operation. RESULTS: Satisfactory results were seen in all the patients with a cosmetically fine appearance. One patient had a urinary tract infection and another, urethrocutaneous fistula and both were amply treated. No glanular adhesion or stenosis occurred. A long-term follow up of 6 months to 4 years (mean: 2 years) in 15 patients did not find any complication. CONCLUSION: The modified preputial island flap urethroplasty technique is an easy, reliable and effective approach to reduce orifice stenosis in hypospadias repair. PMID- 12778330 TI - Absence of sperm meiotic segregation error of chromosomes 1, 9, 12, 13, 16, 18, 21, X and Y in a case of 100% necrozoospermia. AB - Varying degrees of necrozoospermia are common findings in cases of male sub fertility; however, it is rare to find persistent and 100 % necrozoospermia. A case of persistent 100 % necrozoospermia was presented in this paper, where aneuploidy analysis was carried out on sperm. No known associations like thyrotoxicosis, genital infection, spinal injury and diabetes were found. Sperm fluorescent in situ hybridization (FISH) was carried out to evaluate sperm aneuploidy for chromosome 1, 9, 12, 13, 16, 18, 21, X and Y and did not show any excess of aneuploidy over controls. To the best of our knowledge, this is the first attempt on meiotic segregation analysis on 100 % necrozoospermic patients. PMID- 12778332 TI - Molecular characterization of the target antigens of anti-glomerular basement membrane antibody disease. AB - The abnormal immune response to renal antigens is a significant cause of progressive glomerulonephritis and end-stage renal disease, leading to the need for dialysis or kidney transplantation. Type IV collagen of the glomerular basement membrane (GBM), an important component of the blood filtration barrier, is the target of pathogenic antibodies in two forms of anti-GBM antibody nephritis. Type IV collagen is a family of six chains that assemble into three networks with distinct composition and tissue-specific distribution. The GBM contains an alpha3.alpha4.alpha5(IV) network essential for the maintenance of kidney ultrafiltration function: the absence of this network in patients with Alport's syndrome leads to progressive glomerulonephritis. In some Alport patients that receive a kidney transplant, anti-GBM alloantibodies develop against the non-collagenous (NC1) domains of the alpha3.alpha4.alpha5(IV) collagen network, which is present in the renal allograft but absent in the Alport kidneys, causing Alport post-transplant nephritis. In Goodpasture's (GP) syndrome, anti-GBM autoantibodies target the NC1 domain of the alpha3 (IV) chain in the GBM, causing rapidly progressing glomerulonephritis. The GP epitopes have been localized to two homologous regions of the alpha3 NC1 domain, E(A) and E(B), and several populations of autoantibodies with distinct epitope specificity were purified and characterized. The epitopes of GP autoantibodies are sequestered in the NC1 hexamer that connects two adjoining triple-helical molecules. Hydrophobic amino acids have been identified in the epitope of the immunodominant GP(A) autoantibodies, suggesting that the cryptic nature of the GP epitopes is due to interactions among NC1 domains in the NC1 hexamer. Experimental anti-GBM nephritis can be induced in animal models by passive transfer of anti-GBM antibodies or by active immunization with NC1 domains of the alpha3.alpha4.alpha5(IV) network. PMID- 12778333 TI - The role of antibodies and B cells in the pathogenesis of lupus nephritis. AB - Nephritis occurs commonly in lupus patients, and many immunological and nonimmunological factors contribute to disease expression. It is generally appreciated that glomerular immune deposit formation is an early and initiating event, although the mechanisms leading to the deposition of nephritogenic antibodies continue to be debated. Furthermore, it has recently become evident that autoantibodies expressed on B cells also play an important role in pathogenesis. This review focuses on the properties of nephritogenic autoantibodies, their mechanisms of immune deposit formation, and the contribution of B cells expressing autoantibodies to lupus nephritis. PMID- 12778334 TI - T cells in glomerulonephritis. AB - The involvement of immunoglobulin and complement in glomerulonephritis has been recognized for many decades, but the involvement of sensitized T cells and the contributions of cellular immunity have only recently been appreciated. The relative contributions of humoral and cellular immunity in the immunopathogenesis may be an important determinant of the various histological patterns and clinical features of human glomerulonephritis. Subsets of T helper cells, Th1 and Th2, induce immune activation with distinct patterns of involvement of immunoglobulin isotypes and cellular immune effectors. The relative activation of Th subsets is determined by a variety of factors including the nature, dose and mode of presentation of antigens and the cytokine milieu. The balance of Thl/Th2 activation may be important in directing effector pathways and patterns of injury in glomerulonephritis. In the effector phase, T cells are most prominent in proliferative and crescentic glomerulonephritis, where they are usually associated with other mediators of delayed-type hypersensitivity including macrophages and fibrin. In anti-neutrophil cytoplasmic antibody-associated crescentic glomerulonephritis, T cells are invariable participants, frequently in the absence of glomerular antibody deposition. In these forms of glomerulonephritis, which characteristically have a rapid course and poor outcome, T cells are likely to play a pivotal effector role. PMID- 12778335 TI - Role of complement and complement regulatory proteins in glomerulonephritis. PMID- 12778336 TI - Chemokines: therapeutic targets for autoimmune and inflammatory renal disease. PMID- 12778337 TI - Role of nephrin in proteinuric renal diseases. PMID- 12778338 TI - Cell cycle control in glomerular disease. AB - The sequential activation of the cyclin-dependent kinases by their partner cyclins underlies the progression of the cell cycle from quiescence through growth to cell division. More recently a role for these proteins and their inhibitors has been appreciated in several diverse renal and non-renal cell processes, including proliferation, development, differentiation, hypertrophy and apoptosis. The glomerulus represents a unique micro-environment in which to study the cellular outcome following injury, as each of the three resident cell types undergoes a specific and distinct response to a given stimulus. The mesangial cell is capable of marked proliferation, often accompanied by the deposition of extracellular matrix. In contrast, the podocyte has previously been considered a relatively inert cell, and the reparative proliferation of glomerular endothelial cells following injury has recently been described. There is currently increasing awareness of the need to prevent, control and ameliorate the progression of renal diseases. Knowledge of the cell cycle and an understanding of how this may be beneficially manipulated may be crucial to improving the outlook for patients with both diabetic and non-diabetic glomerular disease. PMID- 12778339 TI - New insights into mechanisms of fibrosis in immune renal injury. AB - . Renal fibrosis is the final common pathway for many kidney lesions that lead to chronic progressive organ failure. The tubulointerstitial space occupies up to 90% of kidney volume, indicating that pathological changes in that space can not be without functional significance. By analogy to wound healing, renal fibrogenesis can be divided arbitrarily into three phases: induction, inflammatory, and post-inflammatory phases. The latter phase is of particular importance, since its length often exceeds what would be required for healing. The induction phase is characterized by the infiltration of the tubulointerstitial space by mononuclear inflammatory cells. This influx is mediated by proinflammatory cytokines and chemokines often secreted by activated tubular epithelial cells. Subsequently, these infiltrating mononuclear cells stimulate a heterogeneous group of resident fibroblasts and tubular epithelial cells to undergo phenotypic conversion into activated fibroblasts that secrete abundant extracellular matrix. Tubular epithelial cells contribute to this process through epithelial-mesenchymal transition. During the inflammatory phase these activated fibroblasts are stimulated to produce collagenous matrix mainly by cytokines, such as TGF-beta1, EGF, ET-1, and FGF-2, which are secreted by inflammatory and injured somatic cells. Occasionally however, when inflammation subsides, the matrix synthesis in the post-inflammatory phase of renal fibrogenesis continues and may be more dependent on autocrine stimulation from resident renal cells such as remaining tubular epithelium. Eventually, the collagenous matrix of fibrogenesis destroys blood supply and the perimeter of viability for fibroblasts regresses to the point where scars become acellular. PMID- 12778340 TI - New insights into the pathogenesis of IgA nephropathy. Pathogenesis of IgA nephropathy. PMID- 12778344 TI - The safety and efficacy of chicken type II collagen on uveitis associated with juvenile rheumatoid arthritis. AB - PURPOSE: To investigate the safety and efficacy of chicken type II collagen in treating uveitis associated with juvenile rheumatoid arthritis (JRA). METHODS: A prospective dose-ranging (60 and 540 microg) pilot study of orally administered chicken type II collagen in 13 participants, aged 2-18 years, with JRA and uveitis and without prior exposure to collagens. Anterior chamber cells, flare, vitreous haze, visual acuity, and concomitant anti-inflammatory medications were the ophthalmic outcomes. Arthritis outcomes included the American College of Rheumatology (ACR) core set. RESULTS: No serious or related adverse events were reported. Four participants (2 low dose, 2 high dose) experienced improvement in ophthalmic outcome, while two participants (1 in each group) worsened (p > 0.5). According to ACR criteria, six participants showed improvement in JRA. CONCLUSIONS: Although appearing safe, clearly demonstrating the efficacy of this treatment for JRA or uveitis remains a challenge. Based on the results from this pilot study, a large positive treatment effect on uveitis is unlikely. PMID- 12778345 TI - Bcl-2 expression by CD4 T lymphocytes in Vogt-Koyanagi-Harada disease. AB - PURPOSE: To determine whether Bcl-2 is expressed on CD4(+ ) lymphocytes in the aqueous humor (AH) and cerebrospinal fluid (CSF) of patients with Vogt-Koyanagi Harada (VKH) disease, and to determine whether Fas will induce apoptosis of lymphocytes in the CSF. METHODS: The percentages of CD4, CD8, CD45RO, Fas, and Bcl-2 positive T lymphocytes in the AH and CSF of eight patients with active VKH and five healthy controls were determined by flow cytometry. Soluble Fas ligand (sFasL) in the CSF was measured by ELISA. Freshly isolated cells from the CSF were cultured with anti-Fas antibody (Ab) and apoptosis was assessed by the TUNEL method. RESULTS: Fas(+) CD4(+) lymphocytes were the predominant lymphocytes in the AH and CSF of VKH patients. Bcl-2 was strongly expressed in these cells. Soluble FasL was also detected in the CSF. The number of apoptotic cells detected by anti-Fas Ab was not significantly increased in the CSF of VKH patients. CONCLUSIONS: In spite of the high expression of Fas antigen on CD4(+) cells and the presence of sFasL in the CSF, apoptosis was not observed. Bcl-2 expression may contribute to the regulation of apoptosis of inflammatory cells in the CSF of VKH patients. PMID- 12778346 TI - The role of T(H)1 and T(H)2 cytokines in HSV-1-induced corneal scarring. AB - PURPOSE: To determine the relative impact of T(H)1 and T(H)2 cytokines on the induction of corneal scarring. METHODS: BALB/c and C57BL/6 mice were infected ocularly with herpes simplex virus type 1 (HSV-1)-recombinant viruses expressing either IL-2, IL-4, IFN-gamma, IL-12p35, or IL-12p40. Parental virus and recombinant viruses in one group (dLAT2903, HSV-IL-2, HSV-IL-4, and HSV-IFN gamma) contained an intact neurovirulence gene, gamma34.5, while the second set of recombinant viruses (DM33, dbl-IL2, dbl-IL4, dbl-IFNgamma, dbl-IL12p35, and dbl-IL12p40) lacked the gamma34.5 gene. RESULTS: In the presence of gamma34.5, viruses that expressed either IL-2 or IL-4 reduced the severity of corneal scarring in both BALB/c and C57BL/6 mice compared with the parental virus. In contrast, the recombinant virus expressing IFN-gamma was not protective in BALB/c mice, while it exacerbated corneal scarring in C57BL/6 mice compared with the parental or wt McKrae virus. In the absence of the gamma34.5 gene, recombinant viruses expressing IL-2, IL-4, IFN-gamma, IL-12p35, or IL-12p40 did not induce any corneal scarring. CONCLUSIONS: Our results suggest the following: (1). IL-2 and IL-4 are both involved in protection against HSV-1-induced corneal scarring; (2). IFN-gamma is not involved in protection against HSV-1-induced corneal scarring; and (3). the degree of neurovirulence plays a major role in the protection against or induction of corneal scarring. PMID- 12778347 TI - An immunohistochemical study of the 'snowbank' in a case of pars planiti. AB - PURPOSE: To characterize the extracellular matrix and cellular components of a 'snowbank' removed during vitreous surgery for treatment of retinal detachment complicating pars planitis. METHODS: The 'snowbank' was examined using immunohistochemical techniques and a panel of monoclonal and polyclonal antibodies directed against glial fibrillary acidic protein (GFAP), cytokeratin, alpha smooth muscle actin, tenascin, laminin, fibronectin, and collagen types I, II, and III. RESULTS: The 'snowbank' was acellular except on the uveal side where there were cytokeratin-positive retinal pigment epithelial cells. There were no cells positive for the glial cell marker GFAP and the myofibroblast cell marker alpha smooth muscle actin. The extracellular matrix of the 'snowbank' contained tenascin and collagen types I, II, and III. There was no immunoreactivity for laminin and fibronectin. CONCLUSION: These results on the immunohistochemical components of the 'snowbank' may be useful in clarifying the nature of the chronic inflammatory process in pars planitis. They indicate extensive tissue repair and remodeling, leading to major loss of function. PMID- 12778348 TI - Fuchs' heterochromic iridocyclitis: clinical manifestations in dark-eyed Mexican patients. AB - PURPOSE: To describe the clinical features of Fuch's heterochromic iridocyclitis (FHI) in dark-eyed Mexican patients. MATERIAL AND METHODS: A longitudinal, retrospective, and analytical study of the clinical features of FHI patients was performed. It reviewed the clinical charts of all patients with a diagnosis of FHI seen at the Inflammatory Eye Diseases Clinic of 'Dr. Luis Sanchez Bulnes' Hospital between April 1988 and September 1999. RESULTS: Data of 68 patients (136 eyes) were reviewed. One eye was affected in 89.71% of the cases (61 patients). Fifty patients had brown eyes and iris heterochromia was clinically evident in 25%. Mild stellate keratic precipitates were seen in 80% of the affected eyes. Blurring of the iris surface was found in 40 eyes; iris nodules (Koeppe and/or Busacca) were present in half the cases. A cataract was observed in 69% of the affected eyes, 57.7% of which underwent surgery. Ocular hypertension was present in 30.66% of the affected eyes, glaucoma in 4%. CONCLUSIONS: The more common clinical features in this group of patients were fine and stellate keratic precipitates, atrophy of the iris crypts, and iris nodules. Clinically evident iris heterochromia was present in only 25% of the affected eyes. FHI diagnosis in brown-eyed patients cannot rely on iris heterochromy only. PMID- 12778349 TI - The monthly variation in acute anterior uveitis in a community-based ophthalmology practice. AB - BACKGROUND: A significant increase in the number of episodes of acute anterior uveitis was found in December for two consecutive years at a community-based ophthalmology practice. PURPOSE: To evaluate the monthly variation in episodes of acute anterior uveitis (AAU). METHODS: The charts of all patients with AAU in a practice within a community-based multispecialty ophthalmology group in Albuquerque, New Mexico, were reviewed. Seventy-seven patients with acute, self limited, nontraumatic, unilateral, nongranulomatous anterior uveitis over a two year period were identified. RESULTS: Ninety-four episodes of AAU occurred in 77 patients (42 men, 35 women). The median number of episodes of AAU was three episodes per month (range: 0-14 episodes/month). An increased number of episodes was found in December of both years (11 in December 1996 [p < 0.01] and 14 in December 1997 [p < 0.003]). Smaller clusters of AAU were seen in other months, but did not reach statistical significance. CONCLUSIONS: The presence of a seasonal variation in the number of episodes of AAU suggests that environmental factors are important. A search for such factors that increase the risk of AAU during selected periods may be useful for a better understanding of disease pathogenesis. PMID- 12778350 TI - Intensive immunosuppression treatment for central retinal vein occlusion in a young adult: a case report. AB - PURPOSE: To describe a case of severe central retinal vein occlusion (CRVO) in a young patient, in whom intensive immunosuppressive therapy improved the clinical outcome. CASE REPORT: A 35-year-old men presented with a first episode of CRVO in his right eye in 1990. Despite corticosteroids and laser treatment, rubeotic glaucoma developed and the eye had to be enucleated. Seven years later, CRVO developed in the fellow eye, with venous tortuosity and haemorrhages. An extensive systemic workup was unremarkable. Corticosteroids failed to control the clinical situation. Cells were seen in the anterior vitreous. Visual acuity decreased to 2/60. Cyclosporine and azathioprine were added, but did not prevent recurrences. Campath-1H treatment was then started and visual acuity improved to 6/36. In November 2000, visual acuity was 6/24 and haemorrhages had cleared. DISCUSSION: Many authors have proposed a role for inflammation in the pathophysiology of CRVO in young patients. However, there is no general agreement on corticosteroid use in these patients. Our case illustrates that, in some settings, high-dose corticosteroids and intensive immunosuppression might be used successfully to preserve vision. PMID- 12778352 TI - Role of computed chest tomography (CT scan) in tuberculous retinal vasculitis. AB - AIM: To compare computed chest tomography (chest CT) and routine chest X-rays for the detection of pulmonary tuberculosis in patients with presumed tuberculous retinal vasculitis. MATERIALS AND METHODS: Case reports. RESULTS: Three patients underwent a detailed workup for uveitis. Routine chest X-rays showed normal lung fields in all cases with abnormal hilar shadows in one case. Chest CT revealed the presence of active tuberculous mediastinitis in all three patients. CONCLUSIONS: Chest CT revealed the presence, dimensions, and activity of tuberculous mediastinal lymphadenopathy which routine chest X-rays were unable to detect. Thus, chest CT may be the preferred modality for pulmonary evaluation in these patients. PMID- 12778351 TI - Submacular exudates with serous retinal detachment caused by cat scratch disease. AB - PURPOSE: To present submacular exudates as a manifestation of cat scratch disease. METHODS: Report of two cases. RESULTS: The first patient, a 34-year-old man, developed submacular exudates with serous retinal detachment ten days after having axillary lymphadenopathy and fever. The second patient, a 30-year-old woman, developed submacular exudates with serous retinal detachment mimicking central serous chorioretinopathy. Fluorescein angiography revealed late staining of the subretinal lesions in both cases. The lesion resolved spontaneously in the first patient, while sulfamethoxazole and trimethoprim was required for the second patient. Both patients had a positive IgG titer for Bartonella henselae. CONCLUSIONS: Submacular exudates with serous retinal detachment can occur in cat scratch disease. Cat scratch disease should be included in the differential diagnosis of submacular exudates with central serous chorioretinopathy. PMID- 12778354 TI - [18-FDG-PET-findings in children and adolescents with Hodgkin's disease: retrospective evaluation of the correlation to other imaging procedures in initial staging and to the predictive value of follow up examinations]. AB - Today no evidence based medicine analyses exist about the value of positron emission tomography (PET) in children and adolescents with Morbus Hodgkin. The increasing number of registered PET-examinations within the scope of the GPOH-HD 95 trial motivated to analyse the validity of 18-FDG-PET-examination findings in comparison to the conventional diagnostic methods (CT/MRI/ultrasound) and to the patients follow up. 67 PET-primary staging findings and 48 PET-follow up findings of altogether 106 patients from 27 PET-centres were analysed. Concerning the primary staging findings a concordance of 92% of the PET-findings and the findings of the CT/MRI/ultrasound-examinations per localisation was found, but in more than 50% of the patients a discrepancy occurred in at least one of the 9 investigated localisations. The analysis of the PET follow up findings showed a negative predictive value of 94% in regularly examinations (without previous suspicion of relapse), but only a positive predictive value of 25%. In case of relapse suspicion there was a negative predictive value of 83% and a positive predictive value of 76% in PET. A good prognosis is possible to predict from negative PET follow up findings (relapse risk in regularly controls 7%, at relapse suspicion 17%), whereas the probability for a true relapse in positive PET follow up findings is only markedly increased in case of former relapse suspicion (relapse risk in regularly controls 25%, at relapse suspicion 82%). A prospective multicenter PET study should be realized to analyse systematically the value of PET diagnostics in staging and restaging examinations of children and adolescents with Hodgkin's disease, especially to validate the PET diagnostics in exclusion of vital tumor residuals. PMID- 12778355 TI - [Analysis of causes of death during intensive chemotherapy according to treatment protocol AML-BFM 93]. AB - BACKGROUND: During intensive chemotherapy for AML, more than 10% of patients die because of treatment complications but not because of progression of their underlying disease. In order to improve supportive care and to decrease mortality, we analysed the causes of death and their relationship to the cycles of chemotherapy in children undergoing treatment for AML according to the study AML-BFM 93. RESULTS: Thirty-five (7.4%) of a total of 471 patients treated according to protocol AML-BFM 93 died before or within the first 6 weeks after diagnosis (early death). Fourty-nine patients (10%) did not achieve remission, and 18 (4 %) died of therapy-related complications after having achieved remission. In comparison to earlier AML-BFM studies, early mortality was reduced from 13%, 12%, 9% (AML-BFM 78, 83, 87) to 7% (AML-BFM 93, p-trend = 0.03). In contrast, mortality of patients in complete continuous remission (CCR) did not change. Infectious complications, in particular due to bacterial and fungal pathogens, were the main cause of death. One patient died of arrhythmia associated with SIAD. After stem-cell transplantation in first remission, 7 of 51 patients died, mainly because of graft-versus-host-disease and/or infections. The incidence of infectious complications decreased with the number of chemotherapy cycles and was highest during induction therapy. Fatal complications occurred in one patient during maintenance therapy and in one patient thereafter; both patients were in CCR. Another 14 patients died during intensive therapy (before day 150) mostly with a low percentage of blasts, but no haematologic recovery. The cause of death in these children was mainly bacterial infection or invasive aspergillosis, but seldom progression of leukaemia. CONCLUSION: This analysis confirmed the high incidence of fatal infections in children with AML during chemotherapy-induced severe neutropenia. To increase overall survival in children undergoing therapy for AML, we propose (1) to improve the prophylactic and therapeutic measures for haemorrhage and infections, (2) to continue risk-adapted therapy and (3) to treat high-risk patients in specialised centres only. PMID- 12778356 TI - [Differentiated treatment protocols for high- and standard-risk hepatoblastoma- an interim report of the German Liver Tumor Study HB99]. AB - BACKGROUND: Tumor-free survival from hepatoblastoma could be improved to 75 % of all patients by combining surgery with chemotherapy. This figure reaches 90 % for potentially resectable (SR, standard risk) tumors. The outcome of high risk (HR) hepatoblastomas with multifocally disseminating growth in the liver, invasion of large vessels, extrahepatic extension and metastases is still poor, especially since these tumors often rapidly develop resistance against cytotoxic drugs. In the Study HB 99 of the German Society for Pediatric Oncology and Hematology, it is attempted to reach an improved regression and thereby a better prognosis with inauguration of high dose (HD) chemotherapy. This first interim analysis shall evaluate the preliminary results of this strategy. PATIENTS AND METHODS: 53 children with a hepatoblastoma have entered the study. 8 patients were excluded from this analysis because of different or not completed therapy. 10 SR-patients with a small tumor underwent a primary complete (stage I, n = 8) or microscopically incomplete (stage II, n = 2) resection. These received two courses of ifosfamide (3 g/m2), cisplatin (100 mg/m2), and doxorubicin (adriamycin, 60 mg/m2) (IPA). 26 patients with an extended, but potentially resectable tumor (stage III SR) were preoperatively treated with three courses of IPA, followed by a tumor resection and a 4th course of IPA. 9 patients with a HR hepatoblastoma (3 x stage III HR, 6 x stage IV) were treated with two courses of carboplatin (800 mg/m2) and etoposide (400 mg/m2) (CARBO/VP16). In case of tumor response, they received one or two courses of HD-chemotherapy with carboplatin (2000 mg/m2) and etoposide (2000 mg/m2) after sampling of peripheral stem cells, followed by resection of the primary tumor and metastases, whenever possible. IPA therapy was administered in case of inadequate response to these drugs. The preliminary therapy results were analyzed in relation to the post-surgical stages (I-IV) and to the SIOPEL-PRETEXT (Pretreatment extend of disease) -grouping system (groups I-IV, V, P, E, M). Furthermore, the response of HR-hepatoblastoma to CARBO/VP16, the achieved resectability, and the acute toxicity were evaluated. RESULTS: 40 of 45 (89%) of all hepatoblastoma patients are in remission. 34/36 (94%) SR-patients (stage I - III-SR) are tumor-free, two died of therapy complications. 6 HR-patients are tumor-free, one alive with tumor and two died. In relation to the PRETEXT-grouping, a remission was achieved in 4/4 group I, 14/16 group II, 16/16 group III, 5/6 groups I-III, V, P, E, M, and 1/3 group IV tumors. 6/9 HR-hepatoblastomas were good responders to CARBO/VP16, 5 of these are in remission. 3/9 tumors did not respond, only one could be eradicated by a liver transplantation. In 5/9 HR-patients a R0-resection was possible after chemotherapy, in one a R1-resection, one received a liver transplant, and two tumors remained inoperable. In 4/6 cases lung metastases could be completely removed or, in one case, they had vanished in the CT-scan under chemotherapy. These 4 patients remained in remission. The most frequent severe toxicity of CARBO/VP16 concerned leucopenia (23% of courses) and thrombocytopenia (85% of courses). Under HD therapy severe infections (2/7, 28%) and elevation of transaminases occurred. There was no toxic death. CONCLUSIONS: A cure rate of over 90% can be reached by conventional cisplatin and doxorubicin containing chemotherapy and radical surgery in SR-hepatoblastoma. 50-60% of all hepatoblastomas respond to CARBO/VP16. In these cases HD-therapy with these drugs is highly efficient and enables a remission in the majority of advanced and metastasised HR-hepatoblastoma. A larger number of patients and longer follow-up have to confirm these results. Therefore, the study will be continued. PMID- 12778357 TI - [Euthanasia and caring for the dying in pediatric oncology]. AB - During the last 30 years pediatric oncology has developed therapeutic schemes for all kinds of tumors. Nevertheless, a third of the children suffering from malignancies have still to die. Therefore it is necessary to develop concepts, how to deal with the death of children and how to care for them and their families during the dying process, because the responsibility for these children does not end at the point of finishing therapy, but at the time of their death. Especially during this last part of life these children and their families need an extremely intensive care. Since most of the children want to die at home, we must also be able to care for them there, possibly in cooperation with a local colleague. This, of course, requires an adequate therapy against pain which is possible in most cases. The basement for an optimal care is to be very open to the children. If this openness is established right at the beginning of therapy it will later serve to cope with difficult situations. "Never to lie" is the most important principle. If the patients are not left alone during the dying process the claim for a final injection will be an exception. However, if euthanasia is required, it is rather an expression of despair and a cry for help. The application of very high doses of medicine, necessary in order to prevent pain, might lead to a shortening of life time. This is neither killing on demand nor euthanasia. PMID- 12778359 TI - [Presentation of the "Companion"]. PMID- 12778358 TI - [Quantitative and qualitative evaluation of psychosocial care for patients in pediatric oncology and hematology--a multicenter study on behalf of the PSAPOH]. AB - Due to the ongoing activities to introduce the Disease Related Groups into the German health system, the PSAPOH (Psychosocial Working Group of Pediatric Oncology and Hematology) has initiated a multicenter study to evaluate relevant parameters regarding the amount of psychosocial care in children and adolescents suffering from malignant diseases. This study was performed in 26 German hospitals over three months and evaluated the kind of psychosocial interventions, their duration and frequency and, in addition, their quality. The data collected were evaluated according to the diagnosis, the phase of disease when collecting the data, the psychosocial diagnosis and the grade of risk for psychosocial strain (standard strain [S], multidimensional strain [M] and extreme strain [H]). The results showed no significant differences comparing the amount of psychosocial care within the various kinds of hematological-oncological diseases. But, as expected, a significant and statistically proven increase of the frequency and duration of psychosocial treatment was found according to the stage of the disease and the psychosocial strain. The frequency of treatment is increasing up to 70 % comparing groups with standard strain to groups with extreme strain. Summarizing the amount of psychosocial care within the different phases of disease, a calculation of costs will be possible for each patient. PMID- 12778360 TI - [Bottom-up analysis of the case costs of stem cell transplantation and selected chemotherapies]. AB - BACKGROUND: Up to now, reliable data has not been available on the actual costs of treating oncological patients. However, such data material is urgently required in view of the institution of the health service reform with the concomitant introduction of the G-DRG remuneration system. PATIENTS: The medical services and their costs for 66 patients comprising several stays in hospital, part-time hospitalization and outpatient visits to the clinic categorized "establishment of the diagnosis", "stem cell therapy (SCT)" and "chemotherapy" were recorded in parallel to treatment over a time period of three months. It was thus possible to relate a cost volume of 2.7 million euro to more than 2800 nursing days (full and partial hospitalization) and more than 500 outpatient visits. METHODS: All pertinent costs were collated with the various stays/movements of the patients over 100 calendar days. This was very largely possible thanks to a sophisticated costing and setting off of payments within the hospital on the part of the service providers in the hospital in direct allocation to patients and the third-party payers. Additionally, partial and full surveys as well as questionnaires enabling allocation to the individual case as far as possible, especially the staff requirement, were used. A breakdown of the costs in terms of the patient nursing days was only effected in marginal areas such as the "hotel function" or the general administrative costs. A further subdivision of the stays and movements as well as the phases of the stays applied to the service or the treatment progress above and beyond the areas of treatment. In the case of chemotherapy, a distinction was therefore made between "protocol blocks" and "stays due to complications" and in the case of SCT inter alia even individual phases such as "conditioning". RESULTS AND CONCLUSIONS: It is problematical to represent these patients within an DRG case flat rate system because of frequent and very divergent residence periods with regard to services and costs. The multiplicity of treatment sessions is manifested in the compact "establishment of the diagnosis" to determine a therapy protocol which entails very elaborate inpatient measures under hospitalization, in the expensive and individual "stays due to complications" which usually take place between the "chemotherapy blocks", and the expensive long-term care of the SCT patients after the end of the residence limit (GVD). Owing to the small and divergent numbers of cases in Germany (2000 new patients in 50 centers and 330 SCT per year) and per center, this problem cannot be dealt with by means of a quantity compensation argumentation, as is doubtless justified elsewhere. The actual individual case costs of oncological patients would be 166,72 euro; per outpatient contact and covered by 459,30 euro; per day of hospital care and 808,20 euro; per inpatient treatment day (admission plus discharge day calculated separately and comprised stem cell transplantation patients beyond the residence limit) is covered. The level of the currently applicable case flat rate payments for stem cell transplantations and heterologous donors, mismatched heterologous donors and family donors appear appropriate in relation to the GVD. (The true costs of all oncological patients would indeed be even higher if the GOA payment of the ILV did not only relate to a house-internal low pricing of the GOA scoring value, but would also relate to HLA typing, unpaid physician overtime and the costs of the study centers to the individual cases. From 2002, there will be changes consequent on the judgment of the European Court that stand-by is working time, so that more physicians must be employed and there are expensive changes in the infrastructure owing to the need for a GMP clean area laboratory to process stem cells according to the medical products law.) The indeterminate bound-aries between the three treatment categories place in question the dual financing by the Panel Doctor's Association and the health insurance funds and indicate that a total-sum remuneration appears appropriate. PMID- 12778361 TI - Is hypercortisolaemia a factor in chronic fatigue syndrome? PMID- 12778362 TI - New insights into how adipocytes sense their triglyceride stores. Is cholesterol a signal? AB - In recent years, our view of adipose tissue has evolved from a passive sink for energy storage to an active tissue producing multiple molecules acting on various tissues in different aspects of energy homeostasis. The production of adipose derived secretory products is tightly regulated as a function of adipocyte lipid accumulation, but the mechanisms by which fat cells are able to sense the levels of their triglyceride stores still remains largely unknown. This paper reviews new insights into this question taking cholesterol as a potential intracellular signaling molecule. PMID- 12778363 TI - Nutritional regulation of white adipocyte vascular endothelial growth factor (VEGF). AB - White adipose tissue has distinctive angiogenic properties. White adipocytes are capable of producing vascular endothelial growth factor (VEGF) in response to insulin and catecholamines. Recognizing the dual functions of adipose tissue as an endocrine/metabolic organ and fuel storage depot with an inimitable ability to adjust its organ mass, our aim was to determine whether fasting would affect adipocyte VEGF 165 production. Rats were fed ad libitum, then fasted for 24 hours, and then refed after the 24-hour fast. The isolated white adipocytes from each group and blood endocrine/metabolic profiles were examined at each stage, yielding three sets of results. Using cultured adipocytes, fasting caused a two fold increase in VEGF 165 formation compared to fed rats that normalized after refeeding. Likewise, freshly prepared adipocytes manifested a three-fold augmentation in adipocyte VEGF 165 mRNA expression and a 60 % increase the transcriptional regulator hypoxia-inducible factor 1 (HIF-1alpha) that normalized after refeeding. Blood studies revealed the expected fasting-related alterations in glucose, beta-hydroxybutyrate and corticosterone. Plasma VEGF concentrations were attenuated by 26 % with fasting, and did not normalize with refeeding. Multiple linear regression analyses uncovered statistically significant inverse correlations between plasma VEGF and blood beta-hydroxybutyrate or serum corticosterone. Blood glucose, in contrast, correlated directly with plasma VEGF. We will discuss the potential role of enhanced adipocyte VEGF formation during starvation in light of the known actions of this factor on vascular endothelial mitogenesis and permeability. PMID- 12778364 TI - Endothelial cell specific molecule-1--a newly identified protein in adipocytes. AB - Expression of the endothelial cell-specific molecule (ESM)-1 was originally identified in lung and kidney endothelial cells, where its expression is regulated by cytokines. In vitro, ESM-1 interferes with the molecular mechanisms of immune cell migration by binding to adhesion molecules. In this study, we have explored the expression of ESM-1 in isolated human adipocytes and in rat adipose tissue depots. Human primary adipocytes were cultivated after collagenase digestion and used for in vitro incubation studies. Adipocytes were also isolated from different fat depots of Sprague-Dawley rats. Gene expression was quantified by TaqMan RT-PCR using specific human and rat ESM-1 primers. The cellular localisation of ESM-1 was determined by confocal microscopy using a specific antibody. ESM-1 expression in human adipocytes was stimulated by phorbol ester, an activator of protein kinase C, and by retinoic acid, an activator of nuclear receptors. The maximum increase in gene expression was 3.2-fold after 72 h treatment with phorbol ester and 4.6-fold after 72 h treatment with retinoic acid. The highest expression was found in subcutaneous rat adipose tissue - two fold compared to epididymal and six-fold compared to intrascapular brown adipose tissue. As obesity is related to systemic inflammation (examplified by increased circulating levels of C-reactive protein and interleukin-6), the formation of ESM 1 in adipocytes and its activation by protein kinase C may play a role in the regulation of inflammatory processes. PMID- 12778365 TI - Suppression of aquaporin adipose gene expression by isoproterenol, TNFalpha, and dexamethasone. AB - Aquaporin adipose (AQPap) is a putative glycerol channel in adipocytes. It has recently been shown to be upregulated in insulin resistance stimulated by thiazolidinediones and inhibited by insulin. To further clarify regulation of AQPap gene expression, 3T3-L1 adipocytes were chronically treated with various hormones known to influence insulin sensitivity and adipocyte metabolism, and AQPap mRNA was measured by quantitative real-time reverse transcription polymerase chain reaction. Interestingly, treatment of 3T3-Ll adipocytes with 10 micro M isoproterenol, 10 ng/ml TNFalpha, and 100 nM dexamethasone for 16 h inhibited AQPap gene expression by 62 %, 60 %, and 39 %, respectively; angiotensin 2, growth hormone, and triiodothyronine did not have any effect. The inhibitory effects were dose-dependent with significant suppression detectable at concentrations as low as 1 nM isoproterenol, 1 ng/ml TNFalpha, and 10 nM dexamethasone. Furthermore, inhibition of AQPap gene expression could be almost completely reversed by pretreating 3T3-L1 adipocytes with the beta-adrenoceptor antagonist propranolol. Moreover, stimulation of Gs-proteins with cholera toxin and adenylyl cyclase with forskolin and dibutyryl-cAMP dramatically downregulated AQPap mRNA. Taken together, our results suggest that AQPap is an adipocyte expressed glycerol channel selectively regulated and profoundly downregulated by hormones implicated in the pathogenesis of insulin resistance and dyslipidemia. PMID- 12778366 TI - Prolactin receptor signaling during platelet activation. AB - Prolactin is a newly recognized platelet coactivator that functions through potentiation of ADP-induced platelet activation. However, the possible association between hyperprolactinemia and venous thromboembolism (VTE) has not been systematically investigated up to now; prolactin signaling mechanisms in platelets still need to be elucidated. In this study, plasma prolactin levels in healthy subjects and patients with VTE were determined, demonstrating that patients with VTE and no other congenital risk factors had significantly increased plasma prolactin levels. Moreover, prolactinoma patients demonstrated a higher incidence of VTE than the general population. To elucidate the molecular mechanisms for the development of venous thrombosis, prolactin receptor signaling during platelet activation was investigated with a focus on ADP-stimulated G protein-regulated signaling pathways. The short isoform of prolactin receptors was detected on platelets. Signaling through this receptor, although not directly linked to Gq-proteins, substitutes for Gq-protein regulated signaling pathways involved in platelet activation. We identified protein kinase C, a well established signaling molecule in platelet activation, as a target molecule for prolactin signaling pathways in human platelets. Our findings indicate that hyperprolactinemia may be an important novel risk factor for VTE, suggesting that its thrombogenic effect may be mediated through enhanced platelet reactivity. Revealing the molecular mechanisms of prolactin signaling will allow the design of new antithrombotic therapies. PMID- 12778367 TI - Growth hormone-mediated breakdown of body fat: insulin and leptin responses to GH are modulated by diet composition and caloric intake in old rats. AB - This work was performed to elucidate whether growth hormone (GH)-mediated loss of adipose tissue and responses in plasma insulin and leptin are modulated by diet composition. 12-month-old rats were first fed a high-fat (HF) diet or a low-fat (LF) diet for 14 weeks. After that, GH or saline was administered to rat groups that were maintained on either HF or LF diets or that were switched from the HF to the LF diet. All 6 groups had free access to food. One additional saline group was pair-fed with the GH group that was switched from the HF to the LF diet. The caloric consumption of this latter group was also translated to yet another GH group receiving restricted amounts of the HF diet. GH was given in a total dose of 4 mg/kg/d for three weeks. After sacrifice, blood was collected and tissues were excised. In groups injected with saline, the weight of excised adipose tissue was 60 +/- 4.7, 41 +/- 3.8 and 50 +/- 4.5 g in animals that continued with the HF diet, LF diet, or that were switched from HF to LF, respectively. Corresponding figures after GH treatment were significantly (p < 0.05) decreased to 38 +/- 2.7, 30 +/- 2.3, and 31 +/- 2.7 g, respectively. Pair-feeding had no effect, whereas only 26 +/- 3.0 g of adipose tissue was retrieved in rats fed restricted amounts of HF diet while receiving GH. In this group, plasma insulin and leptin were also significantly (p < 0.05) depressed compared with other GH groups, especially to the group fed the unrestricted HF diet (203 +/- 35 vs. 1345 +/- 160 pmol/l and 9.3 +/- 1.2 vs. 31 +/- 4.4 micro g/l). In conclusion, this study shows that GH mediates breakdown of adipose tissue under a variety of dietary conditions, and that induction of hyperinsulinemia can be prevented if GH treatment is combined with restricted feeding of a diet which is relatively low in carbohydrates and rich in fat. This will also promote a fall of plasma leptin. PMID- 12778368 TI - Growth hormone-mediated breakdown of body fat: effects of GH on lipases in adipose tissue and skeletal muscle of old rats fed different diets. AB - Lipid storage and breakdown is mainly controlled by lipoprotein lipase and hormone-sensitive lipase. The aim of this work was to elucidate whether growth hormone mediated loss of adipose tissue involves a concerted action on tissue lipases, and to what degree such events are modulated by dietary regimen. Twelve month-old rats fed first a high-fat diet or a low-fat diet for 14 weeks were injected with saline or growth hormone (4 mg/kg/d) for four days or three weeks in different combinations with either high- or low-fat diets. In adipose tissue, growth hormone generally inhibited lipoprotein lipase and also attenuated the inhibiting effect of insulin on hormone-sensitive lipase activity. Growth hormone treatment combined with restricted high-fat feeding reduced the activity of both lipases in adipose tissue and stimulated hormone-sensitive lipase in muscle. Generally, plasma levels of free fatty acids, glycerol and cholesterol were reduced by growth hormone, and in combination with restricted high-fat feeding, triglyceride levels improved too. We conclude that growth hormone inhibits lipid storage in adipose tissue by reducing both lipoprotein lipase activity and insulin's inhibitory action on hormone-sensitive lipase. We also propose that growth hormone's effects on tissue lipases and blood lipids are modulated by dietary regimen. PMID- 12778369 TI - Release of beta-endorphin by caffeic acid to lower plasma glucose in streptozotocin-induced diabetic rats. AB - The role of alpha 1A -adrenoceptors in the regulation of opioid secretion from the adrenal glands of streptozotocin-induced diabetic rats (STZ-diabetic rats) was examined in an attempt to determine the mechanism of plasma glucose-lowering action of caffeic acid. In agreement with a previous report, we showed that caffeic acid produced a dose-dependent lowering of the plasma glucose concentration in STZ-diabetic rats along with an increase of plasma beta endorphin-like immunoreactivity (BER). These actions of caffeic acid were abolished by pretreatment with WB 4101 or RS 17 056 at doses sufficient to block alpha 1A -adrenoceptors. In addition, naloxone and naloxonazine at doses effective for blocking opioid micro -receptors abolished the plasma glucose lowering action of caffeic acid. Also, unlike that in wild-type diabetic mice, caffeic acid failed to produce a plasma glucose lowering effect in opioid micro receptor knockout diabetic mice. We observed that caffeic acid could enhance BER release from isolated rat adrenal medulla in a concentration-dependent manner; inhibitors of alpha 1A -adrenoceptors such as WB 4101 and RS 1705 abolished this action. Investigations of the signal pathways further supported that activation of alpha 1A -adrenoceptor is responsible for the stimulatory effect of caffeic acid on BER secretion from the adrenal medulla. In the presence of U73312, a specific inhibitor of phospholipase C, the caffeic acid-induced increase of BER was reduced in a concentration-dependent manner, but it was not affected by U73343, the negative control of U73312. Chelerythrine and GF 109203X also diminished the action of caffeic acid at concentrations sufficient for inhibiting protein kinase C. Moreover, bilateral adrenalectomy in STZ-diabetic rats resulted in the loss of this plasma glucose-lowering effect of caffeic acid, and there was no increase in plasma BER with caffeic acid. Therefore, beta-endorphin release from the adrenal gland appears to be responsible for the lowering of plasma glucose in STZ-diabetic rats induced by caffeic acid, through the activation of alpha 1A -adrenoceptors. PMID- 12778370 TI - Effect of parathyroidectomy on bone mineral density in hemodialysis patients with secondary hyperparathyroidism: possible usefulness of preoperative determination of parathyroid hormone level for prediction of bone regain. AB - PURPOSE: To examine longitudinal changes of bone mineral density (BMD) after parathyroidectomy (PTx) in patients undergoing maintenance hemodialysis (HD) with severe secondary hyperparathyroidism (HPT) to determine which factor contributes most to bone changes. METHODS: Fifteen Japanese HD patients who had been refractory to medical therapy were subject to PTx with autotransplantation. We measured BMD by dual energy X-ray absorptiometry (DXA) at the lumbar spine (L2 - 4 BMD) and the distal 1/3 region of the radius (1/3R BMD) at 1, 3, 6, 12, 24, and 36 months after PTx. RESULTS: Baseline Z-score of BMD was markedly low at 1/3R (- 3.07) and slightly low at L2 - 4 (-0.59) in this group. A significant increase in L2 - 4 BMD was observed as early as one month after PTx, which was sustained afterwards. Annual percent changes in L2 - 4 and 1/3R BMD were + 15.6 % and + 6.4 %, respectively. The annual percent changes in BMD at both sites were positively associated with preoperative intact PTH levels (L2 - 4; r = 0.642, p = 0.010, 1/3R; r = 0.884, p < 0.001) and total alkaline phosphatase (ALP) levels (L2 - 4; r = 0.663, p = 0.007, 1/3R; r = 0.858, p < 0.001). Stepwise multiple regression analysis revealed that serum levels of intact PTH and ALP were the best predictors of both percentage and net changes in radial BMD with high determination coefficients (r 2 > 0.8). CONCLUSION: Successful PTx following appropriate supplementation with vitamin D and calcium provides a marked increase in lumbar BMD and a modest increase in radial BMD in HD patients with secondary HPT. Preoperative levels of PTH and ALP are useful for predicting postoperative changes in bone mass. PMID- 12778371 TI - Hyperleptinemia induced by dexamethasone treatment is not the result of leptin mRNA overexpression in old rats. PMID- 12778372 TI - Liver deiodinase activity is increased in adult rats whose mothers were submitted to malnutrition during lactation. PMID- 12778375 TI - Caustic esophageal strictures in children: 30 years' experience. AB - Many children in developing countries continue to sustain caustic esophageal injures. The first line of treatment is dilatation, unless contraindicated, where 60% to 80% success rate is expected. In cases of failure, esophageal replacement is the only hope for achieving normal swallowing. Over the last 30 years, more than 850 cases of esophageal replacement were done in the Pediatric Surgery Department at Ain-Shams University. Three types of replacement were performed, gastric pull-up (75 cases), retrosternal colon replacement (550 cases), and, in the last 12 years, transhiatal esophagectomy with posterior mediastinal colon replacement (225 cases). Complications in the last 475 cases include 10% cervical leakage, 5% proximal strictures, 2% postoperative intestinal obstruction, 1% mortality, and 0.6% late graft stenosis. Colonic replacement of the esophagus is the ideal treatment in cases of caustic esophageal strictures after failure of dilatation. The posterior mediastinal route is shorter, and in long-term follow up results show improved evacuation and less reflux than with the retrosternal route. PMID- 12778376 TI - Human fetal trachea-SCID mouse xenografts: efficacy of vesicular stomatitis virus G pseudotyped lentiviral-mediated gene transfer. AB - BACKGROUND/PURPOSE: Postnatal gene transfer in respiratory epithelium has been inefficient, particularly in submucosal gland cells, the target cells for cystic fibrosis transmembrane regulator (CFTR) gene transfer in cystic fibrosis. The authors hypothesized that fetal tracheobronchial epithelium may be more receptive to gene transfer in that precursor cells of the respiratory epithelium may be more accessible. METHODS: Vesicular stomatitis virus-G (VSV-G) pseudotyped lentiviral vector was first tested in human fetal tracheal organ culture then validated in a human fetal tracheal xenograft model in severe combined immunodeficiency (SCID) mice. Between 7 x 10(7) and 1.6 x 10(8) transducing units of lentiviral vector encoding the transgene LacZ under the control of the human CMV promoter were administered onto the lumenal surface of the xenografts (n = 6). Biopsy specimens were taken from the xenografts at one month (n = 1), 2 months (n = 2), 3 months (n = 1), 6 months (n = 4), and 9 months (n = 1) after vector administration. Analysis of transgene expression was performed on X-gal stained sections. RESULTS: Transgene expression was observed in 20.2% to 99% of the surface epithelial cells (mean, 70.8 +/- 32.3% SD) and in 28.4% to 99% of the submucosal gland cells (mean, 68.5 +/- 26.2% SD) out to 9 months after vector administration in the tracheal xenografts, equivalent to 63 weeks postconceptual age. No staining was seen in the controls. CONCLUSIONS: Excellent gene transfer in human fetal tracheal xenografts after VSV-G pseudotyped lentiviral vector administration, which may result from more accessible target precursor cells during development, suggests the feasibility of fetal gene therapy for the treatment of congenital airway disease such as cystic fibrosis. PMID- 12778377 TI - Gene alteration of intestinal intraepithelial lymphocytes with administration of total parenteral nutrition. AB - BACKGROUND: Total parenteral nutrition (TPN) is associated with sepsis and loss of immune reactivity. The authors previously have shown that changes in the intestinal mucosal immune system--ie, intraepithelial lymphocytes (IEL)--lead to a loss of epithelial barrier function. This may be a mechanism by which bacteria and toxins endanger individuals receiving TPN. To identify altered IEL gene expression during TPN administration, microarray assays were used. METHODS: Mice received oral feeding (control) or TPN for 7 days. Small bowel IEL were separated and retained, RNA purified, and microarray assays performed (Affymetrix system, 12,491 genes). Results were expressed as quantile-normalized trimmed-means. Significance equals a greater than 2-fold change (TPN v control), P <.01 (t test) or greater than 3-fold, P <.05. RESULTS: In the TPN group 88, IEL genes were significantly up regulated and 114 downregulated (v control). Of these genes, 4 were identified to have highest degree of upregulation (FK506-binding protein 5; mannose-binding lectin, metallothionein 1 and 2), 2 were highly downregulated (microsomal epoxide hydrolase 1 and cytochrome P450 1a1). These genes were found to have high potential for immune-modulatory effects. CONCLUSIONS: The observed alterations in IEL gene expression may have an important role in the altered immune response with TPN and may relate to the increase in sepsis with TPN administration. PMID- 12778378 TI - Trophic enteral nutrition increases hepatic glutathione and protects against peroxidative damage after exposure to endotoxin. AB - BACKGROUND: During total parenteral nutrition (TPN), hepatic concentration of the important intracellular antioxidant glutathione (GSH) is decreased. This study sought to determine whether enteral trophic (small quantity) feeding of GSH precursors would increase hepatic GSH levels during TPN and result in decreased peroxidative injury after endotoxin exposure. METHODS: Sprague-Dawley rats received full TPN for 7 days with postpyloric infusions of either (1) amino acid GSH precursors (cysteine, 60 mg/d; glycine, 86 mg/d; glutamate, 31 mg/d; F1); (2) iso-nitrogenous alanine (132 mg/d; F2); or (3) normal saline (SA). Hepatic GSH concentration was measured by gas chromatography/mass spectrometry. In a parallel study, animals were given TPN and either F1 or SA for 7 days, and endotoxin was administered intravenously before death. Hepatic lipid peroxidation and histology were assessed. RESULTS: Hepatic GSH concentration measured 11.7 +/- 0.6 micromol/g in F1. This was significantly higher (P <.001) than in F2 (7.0 +/- 0.8 micromol/g) or SA (5.0 +/- 0.4 micromol/g). F2 and SA were not significantly different. Hepatic malondialdehyde concentration after exposure to endotoxin was significantly higher in SA (10.36 micromol/g +/- 0.65) than in F1 (7.38 micromol/g +/- 0.77; P <.01). All SA animals had histologic evidence of hepatic necrosis, whereas none of the F1 group showed these changes. CONCLUSIONS: Targeted trophic feeding of GSH amino acid precursors during parenteral nutrition markedly increased hepatic GSH concentration. This was associated with decreased lipid peroxidation and enhanced hepatocellular protection after endotoxin challenge. Thus, targeted trophic feedings may aid in the prevention of TPN related liver disease. PMID- 12778379 TI - In utero limb salvage: fetoscopic release of amniotic bands for threatened limb amputation. AB - The natural history of extremity amniotic band syndrome (ABS) is a progression to deformity or amputation. Fetoscopic laser release of amniotic bands in extremity ABS offers the potential to prevent limb amputation. The authors report on 2 patients with isolated extremity ABS who underwent fetoscopic release. Both patients had isolated extremity ABS with compromised extremity perfusion and no other significant structural abnormality diagnosed sonographically. The first patient underwent fetoscopic release of bands of the left wrist, and the second patient underwent lysis of bands around the right wrist and an unsuspected band around the right lower extremity. Both patients had restoration of blood flow by color Doppler to the affected extremity. There were no perioperative complications. In select patients with isolated extremity ABS, early fetoscopic release may prevent amputation and allow improved limb development. PMID- 12778380 TI - Long-term analysis of children with esophageal atresia and tracheoesophageal fistula. AB - BACKGROUND/PURPOSE: For children with esophageal atresia (EA) or tracheoesophageal fistula (TEF), the first years of life can be associated with many problems. Little is known about the long-term function of children who underwent repair as neonates. This study evaluates outcome and late sequelae of children with EA/TEF. METHODS: Medical records of infants with esophageal anomalies (May 1972 through December 1990) were reviewed. Study parameters included demographics, dysphagia, frequent respiratory infections (> 3/yr), gastroesophageal reflux disease (GERD), frequent choking, leak, stricture, and developmental delays (weight, height < 25%, < 5%, respectively). RESULTS: Over 224 months, 69 infants (37 boys, 32 girls) were identified: type A, 10 infants; type B, 1; type C, 53; type D, 4; type E, 1. Mean follow-up was 125 months. During the first 5 years of follow-up, dysphagia (45%), respiratory infections (29%), and GERD (48%) were common as were growth delays. These problems improved as the children matured. CONCLUSIONS: Children with esophageal anomalies face many difficulties during initial repair and frequently encounter problems years later. Support groups can foster child development and alleviate parent isolationism. Despite growth retardation, esophageal motility disorders, and frequent respiratory infections, children with EA/TEF continue to have a favorable long-term outcome. PMID- 12778381 TI - Advanced neuroblastoma impairs dendritic cell function in adoptive immunotherapy. AB - BACKGROUND/PURPOSE: The authors previously described the complete regression of established neuroblastoma (NB) by the adoptive transfer of syngeneic interleukin 12 transduced dendritic cells (DC) from naive mice. However, some malignancies, like NB, abrogate DC immunostimulation. The authors hypothesize that IL-12 transduction of DC from NB-bearing mice will have the same antitumor properties. METHODS: A/J mice (n = 32) with established NB received peritumoral injection of 1 x 10(6) DC (DC, IL-12 DC, day 7 IL-12 DC or day 14 IL-12 DC) on day 7. Tumor growth, phenotype, and ability to induce NK and T cell activity were measured. RESULTS: Vaccination with naive admIL-12 DC resulted in 100% tumor regression and prolonged survival. Transduced DC induced only partial responses in 75% (day 7) and 25% (day 14) of animals. No differences in phenotype or effector cell activation were noted between admIL-12DC in tumor-bearing or naive mice. CONCLUSIONS: IL-12 DC from tumor-bearing animals have a decreased ability to induce antitumor activity against established murine NB. This decreased capacity appears to be related to the duration of exposure to tumor because day 14 transduced DC had less of an effect than day 7 DC, despite similar phenotypes and ability to activate immune effector cells. PMID- 12778382 TI - Restorative proctocolectomy with ileal pouch-anal anastomosis in very young patients with refractory ulcerative colitis. AB - BACKGROUND/PURPOSE: Restorative proctocolectomy with ileal pouch-anal anastomosis (IPAA) is the standard surgical treatment for ulcerative colitis (UC). The purpose of the current study was to determine the outcome of very young patients (< or =10 years of age) with UC undergoing IPAA. METHODS: Between 1978 and 2002, 13 patients 10 years of age or younger underwent IPAA for management of UC at the authors' institution. Charts were reviewed for patient characteristics, and a standardized telephone interview was performed. RESULTS: Average age at diagnosis was 4.0 years (range, 1.0 to 8.4 years), and patients underwent IPAA at a mean of 6.8 years (range, 3.7 to 10.8 years). Pancolitis was present in 100%. The mean follow-up was 9.1 years (1.0 to 16.1 years), the average number of stools per day was 5 (3 to 8). All patients are continent while awake. Pouchitis was documented in 9 patients (75%). All patients or their parents rated the outcome of their procedures as "excellent." CONCLUSIONS: When compared with older children, very young patients with UC tend to have more frequent total colonic involvement and a greater frequency of pouchitis after IPAA. The functional outcome and patient/family satisfaction with the procedure endorse IPAA as an attractive procedure even in the very young population with UC. PMID- 12778383 TI - A serum factor(s) after small bowel resection induces intestinal epithelial cell proliferation: effects of timing, site, and extent of resection. AB - BACKGROUND/PURPOSE: After small bowel resection (SBR), serum induces proliferation in rat intestinal epithelial cells (RIEC-6). This study was designed to elucidate the effects of postoperative time interval, site, and magnitude of SBR on RIEC-6 proliferation. METHODS: Serum was collected from rats at various times after a 75% mid-SBR or sham operation and added to RIEC-6 cells and growth determined over 5 days. In other experiments, cell growth was recorded in the presence of serum from rats after 25%, 50%, or 75% SBR, or after jejunal or ileal SBR. RESULTS: SBR serum enhanced RIEC-6 cell proliferation as early as 12 hours after resection. The extent of SBR directly correlated with the level of adaptation; however, the effects on cell growth by the serum were similar. SBR serum induced proliferation equally after either proximal or distal resection. CONCLUSIONS: Serum contains a factor that stimulates intestinal cell proliferation soon after SBR but independent of the degree or site of intestinal resection. Although humoral factor(s) play a role in the early induction of enterocyte proliferation after SBR, further modulation of adaptation to varied lengths or sites of intestinal resection are probably governed by mechanisms independent of factors that circulate in the serum. PMID- 12778384 TI - EGF receptor signaling affects bcl-2 family gene expression and apoptosis after massive small bowel resection. AB - BACKGROUND: After massive small bowel resection (SBR), enterocyte apoptosis is elevated and inversely correlates with epidermal growth factor receptor (EGFR) signaling. The purpose of the current study was to determine whether EGFR manipulation affects the expression of specific bcl-2 family members. METHODS: A 50% proximal SBR or sham operation was performed in 3 groups of mice control, after exogenous EGF, or mutant mice with defective EGFR signaling (waved-2). Apoptotic index (no. of apoptotic bodies per crypt), and bax (pro-apoptosis) and bcl-w (anti-apoptosis) protein expression was measured in the remnant ileum after 12, 24, and 72 hours. RESULTS: Waved-2 mice with defective EGFR showed the greatest increase in apoptosis and altered the ratio of bax to bcl-w in favor of apoptosis after SBR. Conversely, EGF prevented the expected increase in apoptosis after SBR by shifting the ratio of bax to bcl-w in favor of cell survival. CONCLUSIONS: After massive small bowel resection, inhibition of the EGFR accelerates the rate of apoptosis and modifies the expression of specific bcl-2 family members to favor apoptosis. These results further support a specific mechanistic pathway for the regulation of enterocyte apoptosis after SBR via EGFR signaling. PMID- 12778385 TI - Serial transverse enteroplasty for short bowel syndrome: a case report. AB - The patient is a 2-year-old boy born with gastroschisis and midgut volvulus that left him dependent on total parenteral nutrition (TPN). At 11 months of age, a Bianchi procedure was performed increasing the total length of bowel from 72 cm to 130 cm. Although he appeared to have sufficient bowel length, he continued to have malabsorption and could only tolerate 10% of his caloric requirement enterally. A barium study found significant dilatation of the lengthened small bowel. At 23 months, we performed a novel bowel lengthening procedure that we have reported previously in an animal model. The serial transverse enteroplasty (STEP) operation increased the 83 cm of dilated and previously lengthened bowel to 147 cm, making the total small bowel length 200 cm. The patient tolerated the procedure well and began to have semisolid bowel movements. Small intestinal absorptive capacity measured by D-xylose absorption showed a substantial increase from 5 to 12 mg/dL (normal range, >20), implying improved but not completely normal small bowel function. This case shows that the STEP procedure increases intestinal length, can be used after a prior Bianchi, and may result in improved intestinal absorptive capacity. The STEP procedure should be considered a surgical option for children with short bowel syndrome. PMID- 12778386 TI - The respiratory advantage of laparoscopic Nissen fundoplication. AB - BACKGROUND/PURPOSE: Laparoscopic Nissen fundoplication is replacing the open approach in the treatment of children with gastroesophageal reflux. The postoperative respiratory advantages seem obvious but remain unproven. The authors hypothesized that laparoscopic Nissen fundoplication provides postoperative respiratory advantages in neurologically normal children as well as those with mental retardation or profound neurologic impairment. METHODS: The charts of all laparoscopic Nissen fundoplications over a 4-year period were reviewed. Sixty-one laparoscopic procedures were compared with the most recent 61 consecutive open Nissen fundoplications. The following variables were reviewed: age, weight, gender, preexisting comorbidities, operating time, postoperative pulmonary complications, and length of stay. Categorical data were compared for significance utilizing chi2 cross tabulation. Variables representing numerical data were compared by t test. RESULTS: Although there appeared to be a trend toward sicker patients in the open group, the laparoscopic group showed significantly improved rates of extubation, shorter recovery room stays, shorter durations of chest physiotherapy, fewer intensive care unit admissions, more rapid resumption of baseline feedings, and overall decreased length of stay (P < 0.05). Pulmonary benefits also were noted in the neurologically impaired population when analyzed separately. CONCLUSIONS: Laparoscopic Nissen fundoplication confers a definable benefit with a significant pulmonary advantage in both neurologically normal children and those with neurologic impairment. PMID- 12778387 TI - Role of immunologic costimulatory factors in the pathogenesis of biliary atresia. AB - BACKGROUND: The authors studied the patterns of expression of immunologic costimulatory molecules (B7-1, B7-2, and CD40) in biliary atresia (BA) patients to confirm any correlation with clinical course/outcome. METHODS: Based on clinical status 2 years postoperatively, 24 BA patients were divided into group I (n = 8, normal liver function), group II (n = 10, anicteric with moderate liver dysfunction), and group III (n = 6, icteric with severe liver dysfunction). Liver biopsies obtained at portoenterostomy and from 6 age-matched controls, were analyzed immunohistochemically using antibodies against B7-1, B7-2, and CD40. RESULTS: There was no expression of B7-1, B7-2, or CD40 in any control liver specimen. In all BA specimens, B7-1, B7-2, and CD40 were expressed strongly in bile ductules in portal tracts. In groups with liver dysfunction, B7-1, B7-2, and CD40 were expressed strongly on the surfaces of Kupffer and dendritic cells and in hepatocyte cytoplasm. Positive staining cells were significantly fewer in patients with better clinical outcome. B7-1 was found in vascular and sinusoidal endothelial cells only in cases of postoperative portal hypertension. CONCLUSIONS: Costimulatory factors expressed on bile ductules, hepatocytes, and vascular endothelial cells appear to mediate autoimmune processes causing progressive liver fibrosis and portal hypertension in BA. PMID- 12778388 TI - Outcome of children with cystic partially differentiated nephroblastoma treated with or without chemotherapy. AB - BACKGROUND/PURPOSE: Cystic partially differentiated nephroblastoma (CPDN) is a rare variant of Wilms' tumor thought to be more favorable than standard nephroblastoma. The purpose of this report is to examine the outcome of children with CPDN, after nephrectomy, treated with vincristine and dactinomycin based chemotherapy (+/- doxorubicin) or no chemotherapy. METHODS: Patients were registered with the National Wilms' Tumor Study Group (NWTSG) and data were collected prospectively. All patients had central review by the NWTSG Pathology Center to confirm the diagnosis of CPDN. RESULTS: Twenty-one patients were identified with a diagnosis of CPDN. Thirteen patients received chemotherapy, and 8 patients did not. In the chemotherapy group the stage distribution was as follows: stage I (n = 10), stage II (n = 2), stage V (n = 1). In the no chemotherapy group, all 8 patients were stage I. All patients had complete tumor resection. There were no cases of disease progression or recurrence in any patient. In patients receiving chemotherapy, 30% (n = 4) had toxicities causing dose reduction. CONCLUSIONS: The outcome of patients with CPDN is favorable with 100% survival rate and no recurrences. For stage I patients, treatment with complete tumor resection appears to be as efficacious as nephrectomy plus chemotherapy. Stage II patients also have excellent outcome when treated with tumor resection and postoperative vincristine and dactinomycin. PMID- 12778389 TI - Neuroblastoma cells inhibit the immunostimulatory function of dendritic cells. AB - PURPOSE: Dendritic cells (DC) are critical for induction of antitumor immunity. Recent studies suggest that tumors may avoid immune destruction by inhibiting DC function. The authors investigated the effect of neuroblastoma (NB) on surface antigen expression and T cell activation by DCs. METHODS: DCs were generated in the presence of granulocyte and macrophage colony-stimulating factor (GM-CSF) and interleukin 4 (IL-4) from peripheral blood of healthy donors. On day 6 of culture, DCs were exposed to human NB cells and were analyzed by flow cytometry. RESULTS: The proinflammatory cytokine tumor necrosis factor alpha (TNF-alpha) failed to upregulate the expression of HLA-DR and costimulatory molecule CD86 by DCs that were cultured with NB. Conversely, upregulation was preserved when DCs were cultured in the absence of NB. Exposure to NB also led to apoptosis of DCs as shown by 2-fold increase in surface phosphatidylserine. It appears that direct contact was required to inhibit DC maturation, because DCs separated from NB cells using a transwell insert did not suppress surface antigen expression. Finally, DCs exposed to NB inhibited the proliferation of allogeneic T cells in mixed lymphocyte reactions. CONCLUSIONS: These findings have significant implications for tumor-pulsed DC vaccines in the treatment of NB and suggest a mechanism by which NB escape rejection. PMID- 12778390 TI - Delayed blind-pouch apposition, guide wire placement, and nonoperative establishment of luminal continuity in a child with long gap esophageal atresia. AB - Delayed primary esophago-esophagostomy is the most physiologically attractive approach to establish luminal continuity in children with pure esophageal atresia. However, excessive tension on the mobilized esophageal ends can lead to anastomotic separation and severe leakage. To circumvent these complications, the blind pouches of a 2(1/2)-month-old child were approximated unopened when, during the extrapleural thoracotomy, a conventional anastomosis was not feasible. A suture was passed through the tip of the 2 ends and made to exit through the mouth and the gastrostomy. After healing occurred, the suture was replaced under fluoroscopic control by a Seldinger-type wire. A vein dilator then was passed over the wire, establishing the continuity between the esophageal segments. Subsequent dilatations increased the anastomosis to the desired diameter. Leakage and an additional thoracotomy were avoided, and the hospital stay was shortened. The child, now 6 years old, is well and swallows normally. PMID- 12778391 TI - The effect of neoadjuvant chemotherapy and surgery in children with malignant germ cell tumors of the genital region: a pediatric intergroup trial. AB - PURPOSE: This study was designed to evaluate (1) the efficacy of standard or high dose cisplatin with etoposide and bleomycin and (2) the role of surgical resection in infants and children with malignant germ cell tumors (MGCT) of the genital region. METHODS: Fourteen of 317 children enrolled in to the Pediatric Oncology Group/Children's Cancer Group intergroup study of MGCT from 1990 through 1996 had genital tumors. Thirteen were eligible for inclusion (12 vaginal, one penile). The initial procedure was biopsy in 11 and subtotal resection in 2. Patients were assigned randomly to receive 4 cycles of etoposide, bleomycin, and either standard or high-dose cisplatin. RESULTS: Nine children underwent postchemotherapy excision of the residual site, and 2 had subsequent biopsies to confirm a complete response. Two with relapse were saved with additional therapy, and one with progressive disease died. The 4-year event-free survival rate in these patients is 76.2% +/- 13.1%, and 4-year survival rate is 91.7% +/- 8.4%. CONCLUSIONS: The author conclude that: (1) the current survival rate for genital MGCT is excellent, (2) delayed surgical resection with organ preservation is not associated with an adverse outcome, and (3) the treatment comparison of the effect of cisplatin dose was inconclusive in this small study population. PMID- 12778392 TI - Living-related partial bladder transplantation for bladder augmentation in rats: an experimental study. AB - PURPOSE: To assess the feasibility of transplanting adult bladder tissue to its offspring as a source of neobladder tissue for bladder augmentation. METHODS: The dome of the bladder of an adult Lewis rat was excised and transplanted into the omentum of a 6-week-old offspring (living-related partial bladder transplant: n = 15). The bladder remnant of the donor rat was closed. Two weeks after transplantation, a laparotomy was performed to mobilize the bladder graft with its omental pedicle into the pelvis. Bladder augmentation (BA) was performed by anastomosing the graft to the recipient's bladder. Thirty days after BA, the entire neobladder was excised and histopathologically examined. RESULTS: At laparotomy, each bladder graft appeared macroscopically as a thin-walled cyst in the recipient's omentum. Each graft could be mobilized into the pelvis and anastomosed to the recipient's bladder. BA was successful in all 15 recipients, and histopathologic studies showed that the mucosa was normal throughout each neobladder. Postoperatively, donors and recipients were clinically well without any sign of urinary incontinence or obstruction. CONCLUSIONS: This is the first report of adult tissue being transplanted successfully into a recipient without vascular reconstruction in a rat. Living-related partial bladder transplantation for the purpose of BA is feasible using our technique and could have application as an alternative technique for BA in a rat. PMID- 12778393 TI - A pilot study of the impact of surgical repair on disease-specific quality of life among patients with pectus excavatum. AB - BACKGROUND: This study was conducted to determine the ability of 2 questionnaires (ie, child and parent versions) to measure physical and psychosocial quality-of life changes after surgical repair of pectus excavatum. METHODS: The authors administered these questionnaires by telephone interviews with 22 parents and 19 children (ages 8 to 18) before surgery and 6 to 12 months after repair by the Nuss procedure. RESULTS: The instruments had high test-retest reliability (Rho > 0.6 for all retained questions). Children reported significant improvements in exercise intolerance, shortness of breath, and tiredness. Of 9 questions asking the children how they feel or act about their bodies, all but one question showed significant improvement after surgery. Parents also reported significant improvements in their child's exercise tolerance, chest pain, shortness of breath, and tiredness and decreases in the frequency of the child being frustrated, sad, self-conscious, and isolated. CONCLUSIONS: These questionnaires appear to be more than adequate to measure disease-specific quality-of-life changes after surgery. These data confirm for the first time that surgical repair of pectus excavatum has a positive impact on both the physical and psychosocial well-being of the child. PMID- 12778394 TI - Suboptimal restraint affects the pattern of abdominal injuries in children involved in motor vehicle crashes. AB - BACKGROUND: Both solid and hollow visceral abdominal injuries have been associated with the use of seat belts in children involved in motor vehicle crashes. The relationship between the types of restraint used and the pattern of abdominal injury is unknown. METHODS: A probability sample of restrained children involved in crashes was enrolled in an ongoing crash surveillance system (1998 through 2002) linking insurance claims data to telephone survey and crash investigation data. Significant abdominal injuries were considered when the Abbreviated Injury Scale (AIS) score was > or =2 and were defined as hollow visceral (HV; intestine, bladder), or solid visceral (liver, spleen, pancreas, kidney). Restraint type was categorized as optimal restraint (OR) or suboptimal restraint (S-OR) based on the child's age and size. RESULTS: For the 33 months of review, interviews were obtained for 13,558 restrained children aged 0 to 15 years, of which, 56% were OR (n = 7,591) and 44% were S-OR (n = 5,967). A significant abdominal injury was recorded in 78 children. A hollow visceral injury was recorded in 38 (9 OR and 29 S-OR), and a solid visceral injury in 32 (18 OR and 14 S-OR). Both hollow and solid visceral injuries were present in 8 children (2 OR and 6 S-OR). Suboptimally restrained children had a higher risk for hollow visceral injury when compared with optimally restrained children (Odds Ratio, 4.14 [95% Confidence Interval 1.33 to 13.22, P <.01]). CONCLUSIONS: Among restrained children with intraabdominal injuries, those who were suboptimally restrained were 4 times more likely to have a hollow visceral than a solid visceral injury when compared with those who were optimally restrained. This suggests that the mechanism of injury for hollow viscus may be directly related to the improper positioning of the restraint. PMID- 12778395 TI - The impact of surgery on infants born at extremely low birth weight. AB - PURPOSE: The aim of this study was to determine the outcome of extremely low birth-weight infants (ELBW) requiring surgical interventions for the complications of prematurity METHODS: One hundred eighty-seven consecutive infants with a birth weight less than 1,000 g treated over a 5-year period were reviewed. Outcome variables included number and types of surgical procedures; length of stay; survival rate and; pulmonary, neurologic, and gastrointestinal morbidity. RESULTS: Surgical interventions were required in 66 (35%) infants (group S) weighing less than 1,000 g at birth (33% necrotizing enterocolitis/bowel perforation, 36% patent ductus arteriosus, 56% other). Overall mortality rate for group S infants was 23% compared with 22% for those not requiring surgery (group NS; P >.05). Mortality rate rose to 38% for those infants undergoing procedures for necrotizing enterocolitis/bowel perforation (P <.05). Although neurologic and pulmonary morbidity for the entire population were high, there was no difference in their incidence between surgical and nonsurgical groups (29% v 26% and 44% v 65%, group S v group NS, respectively; P >.05). CONCLUSIONS: These data suggest an improving outcome for ELBW infants. Common associated morbidities of prematurity do not appear adversely affected by surgical interventions supporting an aggressive approach to the care of these infants at the extreme of life. PMID- 12778396 TI - Slide tracheoplasty for the management of complete tracheal rings. AB - BACKGROUND/PURPOSE: Although rare, complete tracheal rings are the most common cause of congenital tracheal stenosis. The last 2 decades have seen an evolution in management, with increasing awareness of the potential advantages of slide tracheoplasty. METHODS: Between March 2001 and August 2002, 11 children had complete tracheal rings corrected by slide tracheoplasty. Ages ranged from newborn to 15 years, and weight ranged from 1.8 to 57 kg. Length of stenosis ranged from 3 rings to virtually the whole length of the trachea. The most narrow point in the airway varied from less than 1.9 mm to 4.8 mm. Eight children had other congenital anomalies, severe in one child. Most children underwent repair on cardiopulmonary bypass. RESULTS: Nine children are asymptomatic or minimally symptomatic, although 2 have endoscopic evidence of mild residual tracheal stenosis. One child, who had been unstable pre-operatively, died of multiple organ failure 7 weeks postoperatively. A second child required a tracheotomy for bronchomalacia at 6 months and died at 9 months with tracheotomy tube occlusion. Both children had adequate tracheal repairs. Complications have included lateral tracheal stenosis (the "Figure 8" trachea) and recurrent laryngeal nerve damage. CONCLUSIONS: Our management of complete tracheal rings has evolved over the last decade, and slide tracheoplasty currently is our preferred surgical approach for tracheal stenosis regardless of the length of narrowing. PMID- 12778397 TI - Pediatric ileal pouch-anal anastomosis: functional outcomes and quality of life. AB - BACKGROUND/PURPOSE: The aim of this study was to assess and correlate functional outcomes and surgical results with health-related quality of life after ileal pouch-anal anastomosis (IPAA) in pediatric patients. METHODS: Functional outcome was determined by questionnaire and telephone interview. Surgical results were determined by retrospective chart review. RESULTS: Data were gathered from 26 patients (mean age at IPAA, 12 years; mean follow-up, 3.7 years). Diagnoses were ulcerative colitis in 18, indeterminate colitis in 4, and familial polyposis in 4. Indications for IPAA included intractability, medication toxicity, growth delay, and cancer prophylaxis. Short-term complications (5 patients; 19%) included partial small bowel obstruction, stomal revision, pouch abscess, and negative exploration. Long-term complications (8 patients; 31%) were chronic pouchitis and anal stricture. The average number of stools per 24 hours was 3.9. No incontinence was reported; dietary restrictions were negligible. Although there were minimal differences from population norms, parental anxiety remained high. Chronic pouchitis correlated negatively with physical summary score. Nocturnal stooling negatively affected psychosocial quality of life. CONCLUSIONS: Pediatric IPAA resulted in excellent bowel health. Quality of life, physical function, mental health, and self-esteem were equivalent to those of healthy children. These data may help families and physicians make informed surgical decisions. PMID- 12778398 TI - Congenital high airway obstruction syndrome: natural history and management. AB - BACKGROUND: Congenital high airway obstruction syndrome (CHAOS) is a life threatening condition with a poorly understood natural history. METHODS: A retrospective review of five patients with CHAOS between 1997 and 2002 was performed. RESULTS: All fetuses had large echogenic lungs, dilated airways, inverted diaphragms, and massive ascites. One fetus with a laryngeal cyst was terminated at 22 weeks. A twin fetus with findings suggestive of a tracheal web had progressive hydrops, which led to fetal demise. The remaining 3 patients delivered via the ex utero intrapartum treatment (EXIT) procedure survived. The first patient tolerated progressive hydrops for 12 weeks in utero. He had tracheal atresia but underwent laryngotracheoplasty successfully. He is the first long-term CHAOS survivor and is speaking at 5 years of age. The 2 patients with relatively stable lung volumes prenatally have laryngeal atresia with a pinpoint posterior laryngeal fistula. Their postnatal clinical courses were much more benign than the first survivor. CONCLUSIONS: The prenatal natural history and postnatal course of CHAOS depends on whether the airway obstruction is complete. The EXIT procedure offers the potential for salvage of this otherwise lethal condition. Hydrops may be well tolerated prenatally for weeks with potential resolution if airway fistulization is present. PMID- 12778399 TI - The influence of Down's syndrome on the management and outcome of children with Hirschsprung's disease. AB - BACKGROUND: Children with Down's syndrome (DS) have a reportedly poorer outcome after treatment of Hirschsprung's disease (HD) compared with control children. Because of overall improvements in their management, the authors hypothesized that the diagnosis of DS would not influence outcome after the management of HD. METHODS: Consecutive children with HD (1995 through 2002) were collected prospectively then divided retrospectively into those with DS and controls (C). Patients who underwent surgery at another institution and those with total colonic aganglionosis were excluded. RESULTS: Of 66 patients, 9 had DS. Mean age at diagnosis, gender, racial distribution, gestational age, and proximity to our center were similar between groups. Presenting symptoms, location of the transition zone, and type of initial operation were similar. Patients with DS had significantly more comorbidities than controls, which generated significantly greater treatment costs and a higher mortality rate. However, with an average of 22 months of follow-up, the overall outcome including postoperative complications, enterocolitis, and constipation was similar. CONCLUSIONS: These data suggest that in contrast to earlier reports, DS has minimal influence on surgical outcome of patients with HD. Although the overall cost of treating patients with DS is greater, this mainly reflects the impact of managing comorbidities. PMID- 12778400 TI - Transumbilical laparotomy in infants: a novel approach for a wide variety of surgical disease. AB - BACKGROUND/PURPOSE: Laparoscopy has kindled an interest in minimally invasive surgery in infants. To combine improved cosmesis with the advantages of open exposure, we adopted the transumbilical approach for laparotomy in babies up to 26 months of age. METHODS: Only hemodynamically stable babies were considered candidates. The umbilicus was incised nearly circumferentially, and the peritoneum was entered in the midline. The procedure was completed in the standard, open fashion by positioning the incision over the area of interest, and the periumbilical wound was closed subcuticularly. RESULTS: Since 1998, 42 infants have undergone 44 transumbilical laparotomies, with only one conversion to a traditional incision (operating time, 35 to 202 minutes). Twenty-nine newborns were treated transumbilically for duodenal atresia/stenosis/web (n = 11), malrotation (n = 7), intestinal atresia/stricture (n = 5), and other pathology (n = 7). Thirteen older infants (1 to 9 months) were treated transumbilically for intussusception (n = 4), malrotation (n = 4), and other pathology (n = 5). Three toddlers (13 to 24 months) also underwent successful transumbilical laparotomy. There were 2 wound infections and one anastomotic leak. CONCLUSIONS: These results suggest that a transumbilical approach is feasible, safe, and cosmetically superior to the open operation for a broad spectrum of surgical procedures in infants, and it may also be useful in older children. PMID- 12778401 TI - Evaluation of pectus bar position and osseous bone formation. AB - PURPOSE: Minimally invasive repair has become a popular approach for pectus excavatum (PE). The bar is secured to the thoracic wall and left for approximately 2 years. The authors have noticed an intense bone formation (BF) around some of these bars at removal. A review of children undergoing bar removal was performed to better understand this BF in relation to bar placement. METHODS: A retrospective review of children undergoing bar removal after PE repair since January 1998 was performed. Chart review included age at bar insertion and removal, bar insertion position (subcutaneous [SC] v submuscular [SM]), BF on Chest x-ray and at bar removal, operating time, and estimated blood loss (EBL). RESULTS: Thirty-six patients underwent bar removal during the study period (16 SC and 20 SM). Chest x-ray evaluation was possible in 27 patients (16 SM, 11 SC). No difference existed for length of time the bar was in place or age at insertion/removal between groups. EBL was higher in the SM (18.3 v 8.8 mL, not significant). BF was seen radiographically in 15 SM and 3 SC patients (P <.001). BF was encountered at removal in 19 SM patients and a single SC patient (P <.001). Operating time was statistically longer (P <.01) for the SM group (30.2 v 15.6 min). CONCLUSIONS: Bar position during repair of PE is important. SM positioning virtually always results in BF with increased EBL and statistically longer operating time at removal. Careful placement of the bar in the SC position without violating the fascia should be used to avoid these undesirable effects. PMID- 12778402 TI - Primary reversed gastric tube reconstruction in long gap esophageal atresia. AB - BACKGROUND/PURPOSE: Esophageal reconstruction in long-gap esophageal atresia (EA) poses a technical challenge with several surgical options. The purpose of this study was to review the authors' experience with the reversed gastric tube (RGT) in esophageal reconstruction. METHODS: This series describes 7 babies with pure EA treated at 2 centers between 1989 and 2001. Data, gathered by retrospective chart review, included clinical details of the esophageal and associated malformations, technique and timing of repair, early and late complications, and long-term follow-up. Institutional review board (IRB) approval of this study has been obtained. RESULTS: Seven babies were included. Associated malformations were present in 4: trisomy 21 in 2 and imperforate anus in 2. After gastrostomy tube placement, patients were treated with gastrostomy tube feedings and continuous upper pouch suction. Median gap length was 5.5 vertebral segments (range, 3 to 9). RGT with a posterior mediastinal esophagogastric anastomosis was performed at median age of 62 days (range, 38 to 131). There were no anastomotic leaks. Three patients had strictures, one required resection. Exclusive oral nourishment was achieved in 5 patients by 6 months of age. At last follow-up (mean, 4.5 years), 6 patients were receiving oral nutrition exclusively, and all were maintaining growth curves. CONCLUSIONS: In long gap EA, early esophageal reconstruction using an RGT can be performed with minimal morbidity and promising short-term results. PMID- 12778403 TI - Laparoscopic-assisted bowel resection offers advantages over open surgery for treatment of segmental Crohn's disease in children. AB - BACKGROUND/PURPOSE: Minimally invasive approaches have been shown to decrease hospital length of stay (LOS), decrease postoperative pain, and speed return to normal activity for a number of intraabdominal procedures. In this study, laparoscopic (LAP)-assisted bowel resection is compared with an open technique for patients undergoing an initial bowel resection. METHODS: A retrospective review was conducted of 28 patients (12 LAP, 16 open) undergoing initial bowel resection for segmental Crohn's disease. RESULTS: Patients in the LAP group had decrease LOS (5.5 days v 11.5 days) decreased days of parenteral narcotics (3 days v 5 days) and more rapid return to regular diet (3 days v 5 days). CONCLUSIONS: The data suggest that the laparoscopic approach may offer advantages to pediatric patients undergoing an initial bowel resection for segmental Crohn's disease. PMID- 12778404 TI - Mechanism of intestinal-derived fungal sepsis by gliotoxin, a fungal metabolite. AB - BACKGROUND/PURPOSE: Gut barrier dysfunction resulting from fungal overgrowth may be caused by the interaction of gliotoxin (GT), a fungal metabolite, with enterocytes. The goal of this study was to determine the mechanisms by which gliotoxin (GT), a fungal metabolite, causes enterocyte apoptosis. METHODS: The authors measured enterocyte apoptosis, caspase-3 activity, pro-caspase-3, and poly (ADP-ribose) polymerase (PARP) cleavage in GT-exposed IEC-6 cells, a rat intestinal cell line. RESULTS: GT induced apoptosis in IEC-6 cells. The pan caspase inhibitor ZVAD suppressed this GT-mediated apoptosis. GT induced a 15 fold increase in caspase-3 activity over media control. The authors detected PARP cleavage by after GT exposure. DTT pretreatment decreased apoptosis compared with GT alone. CONCLUSIONS: This study supports the concept that fungal overgrowth may lead to gut barrier dysfunction by the local release of gliotoxin and the induction enterocyte apoptosis. PMID- 12778405 TI - Does V-A ECMO increase the likelihood of chylothorax after congenital diaphragmatic hernia repair? AB - BACKGROUND: The authors noticed a relatively large number of patients with congenital diaphragmatic hernia (CDH) repair after extracorporeal membrane oxygenation (ECMO) who had a chylothorax (CT). The data are reviewed. METHODS: The charts of patients from 1990 until 2000 with CDH, treated with or without ECMO, together with the charts of patients treated with ECMO for other reasons and patients with esophageal atresia (EA) repair were reviewed. The diagnosis of CT was made if aspirated fluid appeared chylous and contained more than 90% lymphocytes or if the triglyceride level was more than 1.50 mmol/L. RESULTS: Eighty-nine patients with CDH were analyzed. Postoperatively, 10% had a CT-21% in CDH patients with ECMO treatment and 6% in CDH patients without ECMO treatment. This difference appeared to be significant (P <.05). The presence of a patch as independent variable for the development of CT also showed significance (P <.05). CONCLUSIONS: Chylothorax presented in almost all cases as a left-sided fluid accumulation, and a patch was present in the majority of patients with CDH. Therefore, CT should be considered the result of the severity of the defect rather than the consequence of ECMO as a therapeutic modality. PMID- 12778406 TI - Laryngotracheal separation for intractable aspiration pneumonia in neurologically impaired children: experience with 11 cases. AB - PURPOSE: Laryngotracheal separation (LTS) with or without end-to-side laryngoesophagostomy was performed as an antiaspiration procedure for intractable aspiration pneumonia in 11 children. The effectiveness of LTS for preventing aspiration was investigated. METHODS: Eleven children aged from 9 months to 16 years with intractable aspiration pneumonia underwent LTS with (n = 8) or without (n = 3) laryngoesophagostomy at our institution over the last 2 years. Of these 11 patients, 7 underwent fundoplication with or without gastrostomy for gastroesophageal reflux (GER) before LTS (n = 5) or concurrently with LTS (n = 2). The effectiveness of LTS was evaluated by chart review and follow-up phone questionnaires. RESULTS: LTS decreased the frequency of performing suction from an average of once every 30 minutes to once every 4.5 hours in all patients. In 5 patients who underwent fundoplication with gastrostomy before LTS, aspiration pneumonia remained unless they underwent LTS. Two patients who underwent LTS with or without laryngoesophagostomy tolerated oral feeding postoperatively. All parents rated LTS as excellent or good in terms of improving the quality of life. CONCLUSIONS: LTS can be recommended for neurologically impaired children with intractable aspiration as a primary surgical intervention. If patients show impaired swallowing and GER, LTS could be performed simultaneously with fundoplication and gastrostomy. PMID- 12778407 TI - Distribution of Ca2+-activated K channels, SK2 and SK3, in the normal and Hirschsprung's disease bowel. AB - PURPOSE: The aim of this study was to investigate the expression and distribution of SK2 and SK3 channels in the normal and Hirschsprung's disease (HD) bowel. METHODS: Full-thickness colonic specimens were collected at pull-through operation from 10 patients with HD and from 6 patients during bladder augmentation. Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis for SK2 and SK3 channels and double immunostaining using anti SK2/c-kit, SK3/c kit, SK2/alpha-SMA, and SK2/PGP 9,5 antibodies was performed. Immunolocalization was detected using laser scanning microscopy. RESULTS: RT-PCR analysis showed strong expression of SK2 and SK3 mRNA in the normal human bowel and significantly reduced SK3 expression in the aganglionic bowel (P <.05). In the normal colon, double labeling immunohistochemistry showed strong SK3 immunoreactivity (IR) colocalizing in the c-kit-positive ICCs. In the aganglionic bowel, SK3 IR was reduced markedly in the sparsely found ICCs. There was strong SK2 IR mainly in smooth muscles in the normal and aganglionic bowel. CONCLUSIONS: The results of this study provide the first evidence for the presence of SK2 and SK3 channels and for the immunocolocalization of SK3 channels in the ICCs in the normal human colon. Decreased expression SK3 channels in the aganglionic bowel may contribute to motility dysfunction in HD. PMID- 12778408 TI - Fetal tracheal augmentation with cartilage engineered from bone marrow-derived mesenchymal progenitor cells. AB - BACKGROUND/PURPOSE: The authors have described previously the use of engineered fetal cartilage in a large animal model of fetal tracheal repair. This study was aimed at comparing cartilage engineered from bone marrow-derived stromal cells (BMSC) to native and engineered cartilage, in this model. METHODS: Ovine BMSC were expanded in vitro, seeded onto biodegradable scaffolds, and maintained in transforming growth factor beta 1 (TGF-beta1)-supplemented medium for 3 months (group I). Identical scaffolds were seeded with fetal chondrocytes (group II). All constructs were analyzed in vitro, implanted into fetal tracheas, and harvested after birth for further analysis. RESULTS: There were no differences in survival between the groups. All BMSC-based constructs exhibited chondrogenic differentiation. Matrix analyses in vitro showed that both groups had similar levels of glycosaminoglycans (GAG) and type II collagen (C-II), but lower levels of elastin when compared with native fetal cartilage. Yet, compared with group II, group I had higher levels of GAG, equal levels of C-II, and lower levels of elastin. However, remodeling resulted in no differences between the 2 groups in any of these variables in vivo. CONCLUSIONS: The bone marrow may be a useful cell source for cartilage engineering aimed at the surgical repair of severe congenital tracheal anomalies, such as tracheal atresia and agenesis, in utero. PMID- 12778409 TI - Pediatric surgical images: mixed gonadal dysgenesis. PMID- 12778426 TI - Colonization by Helicobacter pylori (HP) of Meckel's diverticulum (MD). PMID- 12778427 TI - Cases of ileosigmoid knotting. PMID- 12778432 TI - Clostridium difficile colitis associated with hemolytic-uremic syndrome. AB - The authors report the case of a 46-year-old woman who presented with vomiting and profuse bloody diarrhea. Laboratory studies were significant for a hematocrit of 27% and lactate dehydrogenase of 5,394 U/L (5,394 U/L). Her renal function deteriorated rapidly with a peak creatinine of 12.4 mg/dL (1,096.4 micromol/L), and platelet count dropped simultaneously to a nadir of 123,000/microL (123 x 10(9)/L]. Schistocytes were observed in peripheral blood smear. Stool was positive for Clostridium difficile toxin A by enzyme immunoassay (EIA). Stool assay for Shiga-like toxin was negative by EIA, and stool cultures returned negative for Escherichia coli O157:H7 and other enteric pathogens. A diagnosis of C difficile colitis associated with hemolytic-uremic syndrome was made; the patient received plasmapheresis and recovered with no relapse after 10 months of follow-up. This is the second reported case of C difficile colitis associated with hemolytic-uremic syndrome in an adult. PMID- 12778433 TI - Diffuse alopecia in a hemodialysis patient caused by a low-molecular-weight heparin, tinzaparin. AB - The authors report on a 66-year-old hemodialysis patient who had diffuse alopecia while on anticoagulation treatment to prevent extracorporeal blood clotting with a low- molecular-weight heparin (LMWH), tinzaparin. The alopecia reversed completely when tinzaparin was substituted by another LMWH, enoxaparin. The biopsy findings, the possible mechanisms of heparin's action in the hair roots, and the restoration of the patient's hairs after the discontinuation of tinzaparin are discussed. PMID- 12778434 TI - Acute myelo-monocytic infiltrate of the lower esophagus in a 4-year-old renal transplant recipient. AB - BACKGROUND: Acute myeloid leukemia after solid organ transplantation is a rare phenomenon. Few achieve remission; most succumb to relapse and infection. METHODS: A 4-year-old male renal transplant recipient on triple immunosuppression had culture-negative high spiking fever, persistent leukopenia, anemia and severe gastritis. Upper endoscopy showed 2 ulcerating masses in the lower esophagus. RESULTS: Esophageal biopsy showed a highly atypical myelo-monocytic infiltrate. A blastic population of cells featuring convoluted nuclear envelopes with an open chromatin pattern and abundant cytoplasm were filling the submucosa and infiltrating into the muscularis propria. Extensive investigation including bone marrow aspiration showed no spread. Drastic reduction of immunosuppression except 4-mg/d (0.22 mg/kg/d) prednisone for 2 to 3 weeks led to resolution of the leukemic process proven on repeat biopsy. The patient still is in remission 2 years later. CONCLUSION: This case provides evidence that early diagnosis and aggressive reduction of immunosuppression may remit a posttransplant locally invasive acute myelo-monocytic infiltrative process. PMID- 12778435 TI - Zopiclone-induced acute interstitial nephritis. AB - Zopiclone, a relatively new nonbenzodiazepine short-acting hypnotic medication is prescribed frequently for insomnia. The authors report a case of zopiclone induced acute interstitial nephritis in a young, otherwise healthy man. The patient presented with anuric acute renal failure requiring hemodialysis. Kidney biopsy results showed acute interstitial nephritis with a prominent eosinophilic infiltrate in the interstitium. He recovered his renal function after stopping the medication and receiving corticosteroids. PMID- 12778436 TI - Short-term, high-dose pamidronate-induced acute tubular necrosis: the postulated mechanisms of bisphosphonate nephrotoxicity. AB - A 76-year-old man had biopsy-proven acute tubular necrosis (ATN) after intravenous administration of 3 doses of 60 mg of pamidronate (Aredia) over a 2 week period. Pamidronate was given to treat hypercalcemia of unknown etiology. Other potential causes of acute renal failure were excluded with appropriate investigations. The patient's preexisting renal impairment in the context of high doses of pamidronate might have been a potentiating factor for nephrotoxicity. The ATN encountered in this patient resolved; however, short-term hemodialysis was needed. To the best of our knowledge, this is the first reported case of short-term, high-dose pamidronate-induced ATN in the absence of concomitant nephrotoxins. Although necrotic and apoptotic cell death after bisphosphonate administration has been seen in a variety of cells, the exact mechanism of nephrotoxicity is unknown. This report presents a case of pamidronate-induced ATN and discusses the potential mechanisms of bisphosphonate-induced nephrotoxicity. PMID- 12778437 TI - [Spastin gene mutation in Chinese patients with hereditary spastic paraplegia]. AB - OBJECTIVE: To investigate the mutation characteristics of spastin gene in Chinese patients with hereditary spastic paraplegia (HSP) and thus provide a basis for the gene diagnosis of HSP. METHODS: Mutation of spastin gene was screened by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) combined with DNA direct sequencing in 31 unrelated affected HSP individuals in China, of whom 22 were from autosomal dominant families and 9 were sporadic HSP patients. Co-segregation analysis was carried out after the finding of abnormal SSCP bands. RESULTS: Six cases were found to have abnormal SCP bands, and among them, two missense mutations (T1258A, A1293G in exon 8) and one deletion mutation (1667delACT or 1668delCTA or 1669delTAC in exon 14) were found and all of them were not reported previously. They were all co-segregated with the disease and were localized within the functional domain of spastin gene. Besides, T1258A was seen in two unrelated families. CONCLUSION: The mutation rate (18.2%) in autosomal dominant HSP in Chinese patients is comparatively low. Point mutation is the major mutation type and exon 8 may be the mutation hot spot. PMID- 12778438 TI - [Molecular scanning of candidate mtDNA gene fragment in diabetic pedigrees]. AB - OBJECTIVE: To explore novel pathogenic mutation in the mitochondrial DNA gene in diabetic pedigree. METHODS: Twenty-eight suspected mitochondrial DNA diabetic families were recruited. The gene fragment was produced by PCR, and mutation was detected by direct sequencing. RESULTS: In one pedigree, the proband and her mother were found carrying the most common nt3243 A --> G mutation and another 16S rRNA 3205C --> T mutation. But only 3205C --> T was found in her affected brother. All the two patients were deaf and developed diabetes in early age, characterized by impaired beta cell function and low body mass index (BMI). The proband had relatively higher lactic acid concentration than normal individuals. A novel ND1 gene 3434 A --> G(TAT --> TGT) mutation was explored in another proband with deafness and her affected family members. CONCLUSION: 16SrRNA 3205C -> T mutation was found in a mitochondrial diabetes mellitus pedigree, implying its potential pathogenic role in diabetes. Another novel ND1 3434 A --> G mutation was found in another diabetic pedigree. Because this mutation causes amino acid change (Tyr --> Cys) and is co-segregated with diabetes, it may be diabetogenic. PMID- 12778439 TI - [The family investigation of an RHD 270A allele carrier]. AB - OBJECTIVE: Previously the weak D and D category allele were investigated in Caucasian and Japanese families. The current study is aimed at an RHD positive, D antigen negative allele in a Chinese family. METHODS: A pair of primers specific for RHD 270A allele were designed, and a sequence specific primer-PCR (SSP-PCR) method was then established to detect RHD 270A allele in 6 members of a family. Furthermore, RFLP method was used to determine the RHD zygosity in all family members. RESULTS: The RHD 270A allele was detected in the proband, her father and uncle but not grandmother. Therefore this allele may be from grandfather and is inherited through 3 generations. The RHD zygosity test showed that the father and uncle possess one normal RHD gene as RHD 270A carriers, the mother is RHD(+)/RHD( )heterozygote and the individual is RHD 270A/RHD(-)which causes an RHD positive, D antigen negative trait. CONCLUSION: The RHD 270A allele is an ancestral allele, but not a spontaneous. PMID- 12778440 TI - [Molecular cytogenetic detection of partial chromosome 13q trisomy and its relation with the clinical features of tortilcollis]. AB - OBJECTIVE: To search for the possible relation between tortilcollis and partial chromosome 13q trisomy. METHODS: Fluorescence in situ hybridization (FISH) technique combined with chromosome banding was performed to determine the karyotype of two patients with typical clinical features of partial 13q trisomy syndrome, then their manifestations were compared with those of the literatures published previously. RESULTS: The two cases were partial trisomy of 13q14--> ter with a different second derivative chromosome, in spite of this difference, both of them had tortilcollis. CONCLUSION: It is suggested that a potential site for tortilcollis may locate on the long arm of chromosome 13. With reference to a report previously published, the more precise candidate related region may be 13q32--> qter. PMID- 12778441 TI - [Transmission disequilibrium test for congenital dislocation of the hip and HOXB9 gene or COL1AI gene]. AB - OBJECTIVE: To detect the correlation between the congenital dislocation of the hip (CDH) and HOXB9 gene or COL1AI gene. METHODS: A microsatellite DNA marker D17S1820 was chosen in the region of chromosome 17q21 where exists the HOXB9 gene which regulates the embryonic limb development and exists the COL1AI gene. The genotypes of 303 members in 101 CDH nuclear family trios were analyzed by the techniques of polymerase chain reaction(PCR) and denaturing polyacrylamide gel electrophoresis. Then transmission disequilibrium test (TDT) was used to test the data of genotypes. RESULTS: There exist 12 alleles at this polymorphic locus. Transmission disequilibrium was found between CDH and the fourth allele of D17S1820 (chi-square=6.025,P=0.014). CONCLUSION: CDH is associated with the region of chromosome 17q21. HOXB9 gene and/or COL1AI gene may be susceptibility genes of CDH. PMID- 12778442 TI - [Mutational studies of adenomatous polyposis coli gene in carcinomas from patients with hereditary non-polyposis colorectal cancers]. AB - OBJECTIVE: To analyze the mutational features of adenomatous polyposis coli (APC) gene and to explore the effect of mismatch repair (MMR) deficiency on its mutations in hereditary non-polyposis colorectal cancers (HNPCC). METHODS: PCR based in vitro synthesized protein test (IVSP) assay and sequencing analysis were used to confirm somatic mutations of whole APC gene in 19 HNPCC patients. RESULTS: Eleven cases with thirteen mutations were determined. The frequency of APC mutation was 58%(11/19). The exhibiting mutations consisted of 9 frameshift mutations and 4 nonsense ones, indicating the existence of more frameshift mutations (69%). All of frameshift mutations were deletion or insertion of 1-2 bp and most of them (7/9) happened at simple nucleotide repeat sequences, particularly within (A) n tracts (5/9). All of four nonsense mutations resulted from C to T transitions at CpG sites. CONCLUSION: Mutational inactivations of APC gene were detected in more than half of HNPCC patients in this study, indicating that APC mutation is a common molecular event in the tumorigenesis of HNPCC. According to the location of frameshift mutations at simple nucleotide repeat sequences and point mutations at CpG sites, it was suggested that endogenous mechanisms like MMR deficiency might exert an effect on the nature of APC mutations in most HNPCC. PMID- 12778443 TI - [Screening gene mutations of the beta subunit of phosphodiesterase in the Chinese retinitis pigmentosa patients]. AB - OBJECTIVE: To investigate the mutation spectrum of phosphodiesterase beta subunit (PDE6B) gene and incidence in Chinese retinitis pigmentosa (RP) patients. METHODS: Genomic DNA was extracted from the blood samples of 38 patients from 35 autosomal recessive retinitis pigmentosa (ARRP) families and 55 sporadic cases. The mutation of the PDE6B gene was detected using PCR-SSCP, and the amplified PCR product of abnormal bands was sequenced. RESULTS: Within intron 10 of PDE6B gene, a mutation was found in an ARRP family, a G --> A transition at 19th base upstream 5'-end of exon 11. A novel complex heterozygous variant of PDE6B gene in a sporadic case, a T to C transition in codon 323 resulting in the substitution of Gly by Ser and two bp(TG) inserted between the 27th-28th bp upstream of the 5' end of exon 10 were both present in the same isolate RP. But they were not found in 100 unrelated normal individuals. A sporadic RP was found carrying a sequence variant of PDE6B gene, a G to C transversion in intron 18, the 15th base adjacent to the 3'end of exon 18. Another isolate RP was found to have 2 bp inserted between 31st and 32nd base upstream 5'end of exon 4 (in intron 3) of PDE6B gene. CONCLUSION: There is a complex heterozygous mutation of PDE6B gene responsible for a sporadic RP patient in China. Several DNA variants were found in intron of PDE6B gene in the national population. PMID- 12778444 TI - [Polymorphism of two novel SNPs, which locate on chromosome 9p21-22, in Han Chinese of Hunan]. AB - OBJECTIVE: To search novel SNPs in exons and regulatory regions of CDKN2A and two novel putative tumor suppressor genes NGX6 and UBAP1, which all reside on chromosome 9p21-22. METHODS: The exons and regulatory regions of those genes were amplified and sequenced in 96 subjects. RESULTS: Two novel SNPs were found, one resides on the sixth exon of UBAP1 gene and the other on the fourth exon of CDKN2A gene. Two novel SNPs were submitted to the dbSNP database, and their access ID are rs3135929 and rs3088440. The polymorphic information contents of them are 0.102 and 0.213 respectively. There is linkage equilibrium between them, and the polymorphic information content of their haplotype is 0.302, higher than any of them individually. CONCLUSION: The polymorphic information content can be improved by using haplotype analysis of several SNPs. PMID- 12778445 TI - [Association of HMG-CoA reductase gene polymorphism with levels of lipids]. AB - OBJECTIVE: To study the distribution of ScrF1 restriction polymorphism in intron 2 of the 3-hydroxy-3-methylglutaryl coenzyme A(HMG-CoA) reductase gene in Chinese Han population and the association of the polymorphism with coronary heart disease(CHD). METHODS: HMG-CoA reductase genotyping was performed using polymerase chain reaction-restriction fragment polymorphism. RESULTS: HMG-CoA reductase allelic frequencies of A, a were 0.519, 0.481; 0.440, 0.560 in CHD group and control group respectively. There was no significant difference in frequencies of allele and genotype in ScrF1 polymorphism between CHD group and control group(P>0.05). However, the levels of plasma very low density lipoprotein (VLDL) and TG in CHD patients with AA genotype were higher than those in CHD patients with other genotypes(P<0.05). The frequencies of A, a alleles at ScrF1 polymorphic site were significantly different from those reported in European Caucasians (0.44 vs 0.55, 0.56 vs 0.45, P<0.05). CONCLUSION: No direct association was found between the ScrF1 polymorphism and CHD, but there is a significant correlation between the AA genotype of the HMG-CoA reductase gene and the levels of plasma VLDL and TG in CHD group. PMID- 12778446 TI - [Relationship between the Fnu4HI site polymorphism of monoamine oxidase A gene and Parkinson's disease]. AB - OBJECTIVE: To study the association between the polymorphism of human monoamine oxidase type A (MAO-A) gene and Parkinson's disease(PD). METHODS: Fnu4HI restriction fragment length polymorphism(RFLP) and PCR-RFLP were used to detect the mutation of MAO-A gene. The frequencies of alleles and genotypes at the MAO-A Fnu4HI locus on the X chromosome in different PD group were compared with those of the control group. RESULTS: It was found that the frequencies of G allele in the patients with PD and controls were 0.613 and 0.527 respectively, P=0.039 "the frequencies of TT genotype were 0.303 and 0.415(P=0.014), and the frequencies of GG genotype were 0.564 and 0.451 respectively(P=0.021). When the patients were divided into two groups by age-onset, significant difference in the allelic and genotypic frequencies was observed only between early-onset PD group and control group. And when the PD patients were grouped by sex, significant difference was observed only between male PD group and male control group (the frequencies of G allele being 0.669 and 0.500 respectively, P=0.005). CONCLUSION: This study revealed significant differences between PD group and control group in allelic and genotypic frequencies. The findings supported the hypothesis about an association between MAO-A gene and PD, suggesting that age at onset of PD and gender predisposition might be related to the putative association, and Fnu4HI SNP be a risk factor for PD. PMID- 12778447 TI - [The association of paraoxonase 2 gene C311S variant with ischemic stroke in Chinese type 2 diabetes mellitus patients]. AB - OBJECTIVE: To investigate the association between the C311S polymorphism of paraoxonase 2 (PON2) gene and ischemic stroke in Chinese type 2 diabetes mellitus (T2DM) patients. METHODS: A case-control study of 279 Chinese subjects (including 162 T2DM with or without ischemic stroke and 117 non-diabetic control) was performed. Genotype frequencies of C311S polymorphism were studied by polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) analysis with DdeI digestion. RESULTS: C311S polymorphism of PON2 gene was detected in Chinese with the C/S allele frequencies 0.145 and 0.855. The frequency distribution showed significant difference between Chinese and Asian Indian. Furthermore, the genotype distribution (SS, CS and CC) of the PON2 C311S gene polymorphism exhibited a significant difference between T2DM patients complicated with ischemic stroke and T2DM without ischemic stroke, the former had a significantly higher C allele frequency(P<0.05). CONCLUSION: The above data indicate that the polymorphism at codon 311(Cys --> Ser)in the PON2 gene is associated with ischemic morbidity in Chinese T2DM patients and C allele might be a risk factor. PMID- 12778448 TI - [A1166C polymorphism of the angiotensin II type 1 receptor gene and essential hypertension in Han, Tibetan and Yi populations]. AB - OBJECTIVE: To clarify whether A1166C polymorphism of the angiotensin II type 1 receptor (AT(1)R) gene is associated with susceptibility to essential hypertension in Han, Tibetan and Yi populations in China. METHODS: This study involved 302 normotensive and 446 hypertensive subjects. The polymorphism was detected by polymerase chain reaction-restriction fragment length polymorphism in genomic DNA. The data were analyzed by ANCOVA, chi-square test, and multiple logistic regression. RESULTS: In normotensive controls, the A1166 allele frequencies were 0.979, 0.939 and 0.965 in Han, Tibetan and Yi participants, respectively. There was no significant intergroup variation in frequency of the allele in normotensives (chi-square=4.166, P=0.125). The frequency of the A1166 allele in Tibetan male hypertensives was significantly higher than that in normotensives (chi-square=11.46, P=0.001). There was no significant difference in A1166C genotype distribution and allele frequency between normotensives and hypertensives either in the Han (P=0.465) or Yi (P=0.357) populations. Body mass index in the Han and Yi populations (P=0.0001), age in the Tibetan and Yi populations (P=0.0001), and AA genotype in the Tibetan male population (P=0.0034) all were independent risk factors for hypertension. Diastolic blood pressure levels were significantly higher in Tibetan male subjects with the AA genotype than in those with the AC+CC genotype (P=0.0040). CONCLUSION: The A1166 allele is very common in Han, Tibetan and Yi populations, approximately 1.35-fold more common than in Caucasians. The A1166 allele of the AT(1)R gene may be a predisposing factor for essential hypertension in Tibetan males. A1166C polymorphism of the AT(1)R gene is probably not involved in the pathogenesis of essential hypertension in Han and Yi populations. PMID- 12778449 TI - [Coding single nucleotide polymorphism is an ideal marker for detecting gene imprinting by 5' nuclease assay]. AB - OBJECTIVE: To establish a novel approach for quick and high throughput verification of human gene imprinting. METHODS: By use of a pair of dye-labeled probes, 5' nuclease assay was combined with reverse transcriptase-PCR(RT-PCR) to genotype a coding single nucleotide polymorphism (cSNP), rs705(C/T) of a known imprinted gene, small nuclear ribonucleotide protein N (SNRPN), on both genomic DNA and cDNA of human lymphoblast cell lines. RESULTS: Allele discrimination showed a clear monoallelic expression pattern of SNRPN, which was confirmed by RT PCR based restriction fragment length polymorphisms. Pedigree analysis verified the paternal origin of expressed allele, which is in consistency with previous report. CONCLUSION: Coding SNP is an ideal marker for detecting gene imprinting by 5' nuclease assay. This approach has also a potentiality to discover differential allele expression of non-imprinted genes in order to find gene cis acting functional polymorphism. PMID- 12778450 TI - [Diagnosing achondroplasia by single cell nested-PCR]. AB - OBJECTIVE: To research on the reliability of diagnosing achondroplasia (ACH) on single cell level and to provide a basis for preimplantation genetic diagnosis(PGD). METHODS: The high-frequency mutation region G380R of fibroblast growth factor receptor 3(FGFR3) gene was amplified by nested-PCR with single lymphocyte and single blastomere. The products of PCR were digested by restriction enzyme Bfm I, then the digested products were detected by 10% polyacrylamida gel electrophoresis(PAGE). RESULTS: The amplification success rate, allele dropout rate and correct diagnosis rate of single lymphocyte's PCR were 90.4%, 8.2% and 91.8%,respectively. The amplification success rate of single blastomere was 75.4%. CONCLUSION: The diagnosis of ACH by single cell nested-PCR is comparatively stable and reliable. PMID- 12778451 TI - [Gene diagnosis of facioscapulohumeral muscular dystrophy]. AB - OBJECTIVE: To observe the characteristics of changes of p13E-11 labelled 4q35 EcoRI fragments and to make a gene diagnosis of facioscapulohumeral muscular dystrophy(FSHD). METHODS: Genomic DNA was extracted and was digested by EcoR I /Bln I. After pulsed field gel electrophoresis, it was hybridized with probe p13E 11 by Southern blot. The illness was diagnosed as FSHD when the 4q35 EcoRI fragment was smaller than 38 kb. RESULTS: In 26 cases of FSHD, the fragments of 20 cases were smaller than 38 kb. The positive rate was 76.92%. In 12 cases of FSHD family members, the fragments of 2 cases were smaller than 38 kb. All fragments of the 21 controls were greater than 38 kb. CONCLUSION: It was rather good to use <38 kb as a standard for diagnosis of FSHD. The positive rate of FSHD was similar to that from the references. PMID- 12778452 TI - [Study on a new point mutation of nt3426 A --> G of mitochondrial DNA in a diabetes mellitus family]. AB - OBJECTIVE: To elucidate the relationship between point mutations of nt3243A --> G, nt3426 A --> G of mitochondrial DNA and type 2 diabetes mellitus(DM). METHODS: Two hundred patients with type 2 DM and 180 controls with normal glucose tolerance and absence of DM family history were included. The mutations were determined by PCR-restriction fragment length polymorphism. RESULTS: The point mutation nt3426A --> G of mitochondrial DNA ND1 was found in 2 of the patients with type 2 DM (1.0%) but in none of the controls (0). The incidence of this mutation showed no significant difference between the two groups(P>0.05). And none was found to have the mutation of nt3243 --> G. CONCLUSION: The point mutation nt3426 A --> G of mitochondrial DNA ND1 may not be an independent factor to cause type 2 DM. PMID- 12778453 TI - [Dopamine beta hydroxylase gene polymorphism and Parkinson's disease]. AB - OBJECTIVE: To investigate the correlation between the polymorphism of dopamine beta hydroxylase(DBH) gene and the susceptibility of Shanghai Chinese Han population to Parkinson's disease(PD). METHODS: Association study was performed in 144 PD patients and 188 healthy control subjects matched for age, sex and origin. Polymorphism of DBH gene was analyzed with polymerase chain reaction restriction fragment length polymorphism. RESULTS: The allelic frequency of A2 allele of DBH gene was significantly higher in PD patients than in controls(P<0.01).The risk of suffering from PD increased (OR=1.82) in the individual with A2 allele. And the genotypic frequency of A2/A2 was significantly higher in PD patients(OR=2.11, P<0.01),too. On the other hand, the allelic frequency of A1 allele and the genotypic frequency of A1/A2 genotype of DBH gene in PD patients were significantly lower(A1 alleles: OR=0.54, P<0.01; A1/A2 genotypes: OR=0.45, P<0.01). CONCLUSION: The polymorphism in DBH gene might play an important role in the susceptibility of Shanghai Chinese Han population to PD. PMID- 12778454 TI - [Relationship between apolipoprotein E and apolipoprotein B polymorphisms in youths with coronary heart disease]. AB - OBJECTIVE: To investigate hereditary susceptibility to coronary heart disease (CHD) in apolipoprotein E(apo E) and apo B polymorphisms of youths. METHODS: Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to analyze apoE, apoB Xba I, apoB 3' variable number of tandem repeat (VNTR) genotypes for 244 healthy Han students (among them were 109 students with positive CHD family history). RESULTS: The allele frequencies of apo e4, XbaI x(+), 3'VNTR-B(hypervariable element, HVE>38) in the positive group were obviously higher than those in the negative group(P<0.05), and were significantly correlated with the increase in TC, LDL-C, apoB100 levels (P<0.05). CONCLUSION: The alleles for apo e4, XbaI x(+), 3'VNTR-B may be the important genetic markers of Han CHD. PMID- 12778455 TI - [A study on the relation between the apolipoprotein E promoter -427C/T polymorphism and Alzheimer's disease]. AB - OBJECTIVE: To determine the relation between the apolipoprotein E(apoE) promoter 427C/T polymorphism and Alzheimer's disease (AD) in a Chinese Han population in Shanghai. METHODS: The apoE promoter -427C/T polymorphism in 104 AD cases and 110 healthy subjects was detected using polymerase chain reaction method and restriction fragment length polymorphism genotyping technique. The differences in polymorphic distribution between the two groups were tested, and odds ratio was computed. RESULTS: No differences in apoE -427C/T genotypic distribution were observed between AD cases and controls (P>0.05). Even after stratification according to apoE epsilon 4 stratum, there was not any polymorphic distribution difference when epsilon 4 carriers or non epsilon 4 carriers were compared with controls (P>0.05). The association between AD and apoE epsilon 4 appeared in the TT group(OR=3.94,95%, CI:22067038, chi-square=21.48, P<0.05), but not in CT or CC group. CONCLUSION: ApoE -427C/T polymorphism was not a susceptibility factor for AD in this Han population in Shanghai. PMID- 12778456 TI - [The association of HLA-DRB1 allele polymorphism with the genetic susceptibility to liver cirrhosis due to hepatitis B virus]. AB - OBJECTIVE: To assess the association of human leucocyte antigen (HLA)-DRB1 allele with the genetic susceptibility to cirrhosis due to hepatitis B virus(HBV). METHODS: One hundred and six patients with cirrhosis due to HBV in Hubei area were investigated for HLA-DRB1 gene by polymerase chain reaction-sequence specific primers technique. The results were compared with those from 108 normal healthy people. RESULTS: The frequency of HLA-DRB1*1201/1202 allele was 20.28% in patient group, which was significantly higher than the frequency (6.01%) in control group, the relative risk (RR) being 4.9878 (P<0.01). The frequency of HLA DRB1*1501/1502 allele was decreased in patient group (patient 6.6%, control 16.67%, RR=0.3043, P<0.05), while the frequencies of other HLA-DRB1 alleles were not significantly different(P>0.05). CONCLUSION: HLA-DRB1*1201/1202 allele may be the susceptibility gene in patients with cirrhosis due to HBV in Hubei Han nationality; HLA-DRB1*1501/1502 allele is a resistant gene in patients with cirrhosis. PMID- 12778457 TI - [Polymorphisms of six short tandem repeat loci in Zhejiang She ethnic population and Zhejiang Hans of China]. AB - OBJECTIVE: To investigate genetic polymorphism of six short tandem repeat (STR) loci (D3S1358,D16S539, TH01,TPOX, CSF1PO,D7S820) in Zhejiang She ethnic population and Han population. METHODS: By use of AmpFlSTR Cofiler kit, 6 STR loci in 108 She samples and 102 Han samples were amplified. The PCR products were electrophoresed by ABI Prism 377 sequencer; the data were analyzed by Genescan software. RESULTS: All genotype frequencies of the 6 STR loci in She and Han ethnic groups met Hardy-Weinberg equilibrium. In the She population, the heterozygosities (H) in D3S1358,D16S539,TH01,TPOX, CSF1PO and D7S820 were 0.8028, 0.9148, 0.7522, 0.6728, 0.9123,0.8338, the exclusion of paternity(EP) were 0.4067, 0.6057, 0.4437, 0.3200, 0.5250, 0.5358, discrimination power (DP) were 0.6690, 0.7841, 0.6447, 0.5382, 0.7298, 0.7296.The combined DP, PE and polymorphism information content were 0.9991,0.9805,0.9988 respectively. There were significant differences at D3S1358, D16S539 and TPOX loci, compared with Hans. CONCLUSION: She population has its own STR allele distribution characteristic. The above data obtained from She population can be used not only in genetic researches and population investigation, but also in human identity and paternity testing. PMID- 12778458 TI - [Relationship of I/D polymorphism of angiotensin converting enzyme gene with hypertension in Xinjiang Kazakh isolated group]. AB - OBJECTIVE: To investigate whether the insertion/deletion(I/D) polymorphism in the angiotensin converting enzyme(ACE) gene is associated with essential hypertension in Xinjiang Kazakh isolated population. METHODS: The study covered 201 hypertensives and 151 normotensive controls in Xinjiang Barlikun Kazakh population. The I/D polymorphism of ACE gene was determined by polymerase chain reaction. RESULTS: The frequencies of D and I in the hypertensive group (0.44 and 0.56, respectively) were not significantly different from the controls(0.39 and 0.61, respectively, P=0.16). The frequencies of ACE genotypes of DD, ID, and II were 0.18, 0.52, 0.30 in hypertensives respectively and 0.17, 0.43, 0.40 in control group respectively. There was no significant difference in genotypes between hypertensive group and normotensive group (P=0.14). CONCLUSION: The results suggested that the I/D polymorphism of ACE gene might not be associated with hypertension in the Kazakh population of Xinjiang Barlikun area. PMID- 12778459 TI - [DNA mismatch repair enzyme hMSH2 genetic polymorphism in southern Chinese Han population]. AB - OBJECTIVE: To study hMSH2 genetic polymorphism in southern Chinese Han population. METHODS: The basic materials and blood samples from 163 southern Chinese were collected. The mutations of exon 6 and exon 7 of hMSH2 gene were investigated by PCR-SSCP, followed by DNA sequencing. RESULTS: Fragments of 250 bp including exon 6 and fragments of 323 bp including exon 7 of hMSH2 gene were amplified by multiple PCR. The allele frequencies of C18, A82 and B39 type mutations were 0.0184, 0.0031, 0.0031, respectively. The gene frequencies and gene type frequencies of three polymorphism sites in normal population accorded with Hardy-Weinberg equilibrium (P>0.05). The heterozygosity of C18 type mutation (0.0361) was the highest. CONCLUSION: There were three polymorphism sites in exon 7 of hMSH2 gene in southern Chinese Han population, among which the genotype frequency of C18 type was the highest, suggesting that C18 type mutation be a useful genetic mark. PMID- 12778460 TI - [Genetic polymorphism of six short tandem repeat loci in the Han population in Hebei province of China]. AB - OBJECTIVE: This article reports a population genetic study on six short tandem repeat(STR) loci, D7S820, D19S253, D12S391, D5S818, D16S539 and D8S1179, in a sample of unrelated Chinese Han individuals(n=122-173) living in Hebei province. METHODS: DNA extraction from blood samples (200 in number) and multiplex amplification of the above six loci were carried out. Using denaturing polyacrylamide gel electrophoresis and silver stain, the authors investigated the distribution of allele frequencies of the six loci in Han population in Hebei province. RESULTS: The STR polymorphisms at all of the six loci were observed in Chinese Han population in Hebei province. The observed heterozygosities of D7S820, D19S253, D12S391, D5S818, D16S539 and D8S1179 were 0.828, 0.757, 0.769, 0.837, 0.785 and 0.852, respectively. The measured values of the power of discrimination (PD) were 0.914, 0.919, 0.940, 0.909, 0.917, 0.944; of the mean exclusion chance(MEC) 0.618, 0.740, 0.801, 0.557, 0.655, 0.696 and of the polymorphism information content (PIC) in Chinese 0.771, 0.760, 0.762, 0.708, 0.776 and 0.794, respectively. CONCLUSION: The genotype distributions of the six STR were in accordance with Hardy-Weinberg equilibrium. The numerical values of the PD and MEC are relatively high in Hebei province, and thus can be of significant application in population genetics and forensic medicine. PMID- 12778461 TI - [Association study on HLA-DP and -DQ allelic polymorphisms and nasopharyngeal carcinoma in the Han nationality in Hunan province]. AB - OBJECTIVE: To investigate whether the polymorphisms of HLA-DPA1,DPB1,DQA1 and DQB1 alleles were associated with nasopharyngeal carcinoma(NPC). METHODS: Eighty seven NPC patients and 91 normal controls of Han nationality in Hunan province were genotyped for HLA-DPA1, HLA-DPB1,HLA-DQA1 and HLA-DQB1 by PCR/SSO technique. RESULTS: The frequencies of allelic gene DPA1*0201, DPB1*1901 and DQA1*0201 were lower, and of DPB1*0402, DQA1*0101 were higher in patients than in controls; the frequencies of haplotype DPA1*0201-DPB1*1401 and DQA1*0201-DQB1*0201 in patients were lower than those in controls; however, the values of P are not significant after Bonferroni correction(Pc>0.05). CONCLUSION: No significant association between the HLA-DP and HLA-DQ loci and NPC in Han nationality in Hunan province was confirmed. PMID- 12778462 TI - The role of interleukin-12 in human infectious diseases: only a faint signature. AB - IL-12 is the signature IFN-gamma-inducing cytokine and, as such, is thought to be crucial for protective immunity against intracellular microorganisms. This concept is supported by results from experimental infections of knockout mice lacking IL-12 or the IL-12 receptor. The description of human patients with inherited IL-12 or IL-12-receptor deficiency challenges this view. Indeed, in natural conditions of infection and immunity - the hallmark of the human model - IL-12 was found to be redundant in defense against intracellular microorganisms other than Mycobacteria and Salmonella. More surprisingly, IL-12 was recently found to be redundant even in defense against primary intection by Mycobacteria and Salmonella in many patients, and against secondary infection by Mycobacteria but not Salmonella in most patients. PMID- 12778464 TI - Selective absence of CD8+ TCRalpha beta+ intestinal epithelial cells in transgenic mice expressing beta2-microglobulin-associated ligands exclusively on thymic cortical epithelium. AB - Whereas interactions between the TCRalpha beta and self MHC:peptide complexes are clearly required for positive selection of mature CD4(+) and CD8(+) T cells during intrathymic development, the role of self or foreign ligands in maintaining the peripheral T cell repertoire is still controversial. In this report we have utilized keratin 14-beta2-microglobulin (K14-beta2m)-transgenic mice expressing beta2m-associated ligands exclusively on thymic cortical epithelial cells to address the possible influence of TCR:ligand interactions in peripheral CD8(+) T cell homeostasis. Our data indicate that CD8(+) T cells in peripheral lymphoid tissues are present in normal numbers in the absence of self MHC class I:peptide ligands. Surprisingly, however, steady state homeostasis of CD8(+) T cells in the intestinal epithelium is severely affected by the absence of beta2m-associated ligands. Indeed TCRalpha beta(+) IEL subsets expressing CD8alpha beta or CD8alpha alpha are both dramatically reduced in K14-beta2m mice, suggesting that the development, survival or expansion of CD8(+) IEL depends upon interaction of the TCR with MHC class I:peptide or other beta2m-associated ligands elsewhere than on thymic cortical epithelium. Collectively, our data reveal an unexpected difference in the regulation of CD8(+) T cell homeostasis by beta2m-associated ligands in the intestine as compared to peripheral lymphoid organs. PMID- 12778463 TI - Role of Toll-like receptors in costimulating cytotoxic T cell responses. AB - Stimulation of Toll-like receptors (TLR) by pathogen-derived compounds leads to activation of APC, facilitating the induction of protective immunity. This phenomenon is the basis of most adjuvant formulations currently in development. Here, we tested the ability of TLR2, 3, 4, 5, 7 and 9 signaling to enhance CTL responses upon vaccination with virus-like particles. Stimulation of TLR2 and 4 failed to increase CTL responses, whereas ligands for TLR3, 5 and 7 exhibited moderate adjuvant function. In contrast, stimulation of TLR9 dramatically increased CTL responses, indicating that ligands for TLR9 are likely to be the most promising candidates for the development of novel adjuvant formulations for stimulating CTL responses. PMID- 12778465 TI - Impaired contact hypersensitivity in macrophage migration inhibitory factor deficient mice. AB - To determine whether macrophage migration inhibitory factor (MIF) is required for contact hypersensitivity (CHS) response, MIF-deficient (MIF KO) and wild-type (WT) mice were sensitized with trinitrochlorobenzene (TNCB) or oxazolone on their abdominal skin and challenged on the dorsum skin of one ear 5 days later. Significant ear swelling was observed in the WT mice, but this response was inhibited in the MIF KO mice (p<0.01 for MIF KO vs. WT mice in 24 h). In addition, lymph node cells from hapten-sensitized MIF KO mice showed a decreased capacity for transferring the CHS response. A topical application of TNCB (200 microg) caused a significant decline in epidermal Langerhans cell (LC) density (20.3%; p<0.01 compared with vehicle) 4 h after application in WT mice, but it failed to provoke a significant epidermal LC migration in MIF KO mice (7.4%). By mixed lymphocyte reaction, the T cell proliferative response to alloantigen was significantly decreased in the MIF KO mice compared with WT mice (p<0.005). Taken together, these results indicate that MIF is pivotal in the regulation of cutaneous immune responses and plays a central role in LC migration and T cell proliferation for the CHS response. PMID- 12778466 TI - Skin-versus gut-skewed homing receptor expression and intrinsic CCR4 expression on human peripheral blood CD4+CD25+ suppressor T cells. AB - In humans and rodents a population of naturally occurring CD4(+)CD25(+) T cells are suppressor T (CD25(+) Ts) cells, which are considered to maintain peripheral immunological tolerance. Recently, we have described a unique chemotactic response profile for human CD25(+) Ts cells, but their homing potential remains poorly defined. Here, we document a heterogeneous homing potential of human peripheral blood CD25(+) Ts cells consistent with their ability to mediate immunosuppression at distinct locations. Surprisingly, CD25(+)Ts cells are depleted of gut-homing integrin alpha(4) (+)beta(7) (+) T cells, while being enriched in skin-homing cutaneous lymphocyte antigen (CLA)(+) T cells. These findings document heterogeneous homing potential of peripheral blood-borne CD25(+) Ts cells with marked skewing for skin- versus gut-homing. Expression of CCR4 associates with both CD25 and CLA cell surface markers, being highest on CD4(+)CLA(+)CD25(+) T cells. Importantly, CD4(+)CD25(+) Ts cells isolated from human cord blood lack expression of CLA while expressing CCR4, suggesting intrinsic expression of CCR4 on CD25(+) Ts cells. These observations indicate that the increased expression of CCR4, which is proposed to guide CD25(+) Ts cells to DC, is an intrinsic feature of CD25(+) Ts cells. PMID- 12778467 TI - Unique T cell proliferation associated with PKCmu activation and impaired ZAP-70 phosphorylation in recognition of overexpressed HLA/partially agonistic peptide complexes. AB - Altered peptide ligands (APL) induce T cell responses different from those induced by the original agonistic peptide. As shown for CD4(+) T cells, partial agonists induce partial T cell activation without proliferation because of lower affinities and higher off rates to TCR than those of agonists. To determine whether overexpression of partially agonistic TCR ligands on antigen-presenting cells provides high-avidity TCR ligands, we generated L cell transfectants expressing various numbers of HLA-DR4 covalently linked with APL derived from a streptococcal peptide and observed responses of the cognate T cells. Some overexpressed HLA-DR4/partially agonistic APL complexes induced T cell proliferation in a density-dependent manner. However, tyrosine phosphorylation of zeta-associated protein-70 (ZAP-70) and linker for activation of T cells and kinase activity of ZAP-70 were not detectable. T cell proliferation stimulated with L cell transfectants was sensitive to the PKC inhibitor Go6976, but to a lesser extent to Go6983, suggesting the involvement of mu isotype of PKC (PKCmu). In vitro kinase assays revealed that PKCmu activity was up-regulated only in T cells stimulated with L cell transfectants that induced T cell proliferation. Our data suggest the presence of a unique signaling pathway coupling TCR ligation with T cell proliferation associated with PKCmu activation and impaired ZAP-70 activation. PMID- 12778468 TI - Targeting of immunostimulatory DNA cures experimental visceral leishmaniasis through nitric oxide up-regulation and T cell activation. AB - Active targeting of CpG-containing oligodeoxynucleotide (CpG-ODN) to macrophages was studied by incorporating it in mannose-coated liposomes, using visceral leishmaniasis as the model macrophage disease. Mannosylated liposomal CpG-ODN was more effective than liposomal or free CpG-ODN in inhibiting amastigote multiplication within macrophages. Moreover, in a 60-day mouse model of visceral leishmaniasis, complete elimination of spleen parasite burden was achieved by mannosylated liposomal CpG-ODN, compared to 62% and 81% parasite suppression by free and liposomal ODN, respectively, at a similar dose. Although in vitro exposure of CpG-ODN did not induce marked nitric oxide (NO) generation by macrophages, considerably enhanced amount of NO was generated by macrophages of CpG-ODN-treated animals. Their splenocytes secreted soluble factors required for the induction of NO generation, and the increased NO generation was paralleled by an increase in antileishmanial activity. Inducible NO generation was suppressed by treating splenocyte supernatants with anti-IFN-gamma or anti-IL-12 antibodies, whereas in vivo administration of these anti-cytokine Ab along with CpG-ODN reversed protection against infection. CpG-ODN treatment resulted in reduced levels of IL-4, but increased levels of IFN-gamma, IL-12 and inducible NO synthase in infected spleen cells, which was magnified by encapsulation in mannose-coated liposomes. This targeted treatment was not only curative, but it also imparted resistance to reinfection. These results represent a general approach for intracellular targeting of CpG-ODN, which effectively enhances its therapeutic potential in redirecting curative Th1 responses in Th2-driven disorders. PMID- 12778469 TI - Deficiency of CD26 results in a change of cytokine and immunoglobulin secretion after stimulation by pokeweed mitogen. AB - To investigate the role of CD26 in the immune system, CD26 gene knockout mice with C57BL/6 background were used to study the immune response after stimulation with PWM. CD26(-/-) mice display an apparently normal phenotype. However, in their spleen lymphocyte population the percentage of CD4(+) T cells is lower, and that of NK cells is higher, than that in CD26(+/+) mice. In their peripheral blood, CD26(-/-) mice present a conspicuously decreased proportion of CD4(+) NKT lymphocytes. In vitro, the PWM-stimulated IL-4 production was decreased by 60-80% in the supernatants of spleen lymphocytes of CD26(-/-) mice compared to that of CD26(+/+) mice, whereas levels of IL-10 and IFN-gamma were increased. No significant differences were found in the production of IL-2, IL-5, IL-6 and IL 13 between knockout and wild-type mice. After immunization of mice with PWM in vivo, serum levels of total IgG, IgG1, IgG2a and IgE were markedly lower in CD26( /-) mice than those in CD26(+/+) mice, while no difference was found in IgM production. Further analysis of cytokine levels in vivo revealed a reduced IL-4, IL-2 and delayed IFN-gamma production in sera of CD26(-/-) mice upon immunization with PWM. These results indicate that CD26 contributes to the regulation of development, maturation and migration of CD4(+) T, NK and NKT cells, cytokine secretion, T cell-dependent antibody production and immunoglobulin isotype switching of B cells. PMID- 12778470 TI - Human cytomegalovirus impairs dendritic cell function: a novel mechanism of human cytomegalovirus immune escape. AB - Human cytomegalovirus (HCMV) employs multiple mechanisms to evade the immune system and succeeds to persist lifelong in the host. Human dendritic cells (DC) are the main antigen-presenting cells and play the key role in inducing and maintaining immune responses. Here, we studied the interaction of HCMV with DC. We found that DC, irrespectively of their stage of maturation, were fully permissive for HCMV when endothelial cell-adapted HCMV strains were applied. When fibroblast-adapted strains were used, viral replication was abrogated at the level of immediate early (IE) and/or early (E) gene expression. Irrespective of the HCMV strain used, infection of DC prevented the signal delivery essential for T cell activation in a multistep manner. Furthermore, we observed an altered expression of adhesion molecules. This might contribute to an impairment of DC migration. Our data indicate that a soluble factor induced by IE and/or E genes is involved in these processes. The impairment of DC function upon HCMV infection may contribute to virus-mediated immunosuppression and help the virus to establish persistence in the host. PMID- 12778471 TI - Vaccination with myelin oligodendrocyte glycoprotein adsorbed to alum effectively protects DBA/1 mice from experimental autoimmune encephalomyelitis. AB - To prevent an organism from developing autoimmunity the body limits the number of autoreactive cells through thymic negative selection and regulates their activity through induction of suppressor T cells. Development of antigen-specific therapies provides an interesting opportunity to imitate the body's own, often effective, method of protection. Our study demonstrates that DBA/1 mice could be protected from experimental autoimmune encephalomyelitis induced through injection of recombinant myelin oligodendrocyte glycoprotein (rMOG) when they were previously immunized intraperitoneally with rMOG adsorbed to aluminium hydroxide. This protection was associated with a decreased IFN-gamma production by rMOG-specific cells, but not a decreased proliferative response. Protection was long lasting, indicating that MOG-alum vaccination might be developed as a prophylactic therapy in multiple sclerosis. PMID- 12778473 TI - Virus infection expands a biased subset of T cells that bind tetrameric class I peptide complexes. AB - We have used a TCR beta-chain transgenic mouse to examine the relationship between the ability of a T cell to bind soluble class I-peptide complexes and its response to antigenic stimulation in vivo. T cells from gBT-I.3beta TCR beta chain transgenic mice preferentially carried TCR alpha-chains bearing the same Valpha2 V region as found in the parent receptor specific for an immunodominant HSV-1 gB-peptide. Furthermore, CD8(+) T cells from these mice bound K(b)-gB tetrameric complexes with relatively high frequency, and most of these cells contained a Valpha2 TCR alpha-chain. Detailed sequence analysis of the tetramer binding peripheral T cells showed that this was a heterogenous population expressing TCR with only partial sequence similarity to the parent receptor, which took the form of preferential inclusion of the parental Jalpha16 element. Infection with HSV-1, however, selected a subset of tetramer-positive T cells. This was based on the emergence of a co-dominant Jalpha usage and selection of a restricted CDR3alpha length. Therefore, the ability to bind soluble MHC-peptide complexes does not always correlate with the ability of a T cell to respond to its cognate antigen after in vivo stimulation. PMID- 12778472 TI - Efficient mucosal delivery of the HIV-1 Tat protein using the synthetic lipopeptide MALP-2 as adjuvant. AB - A major requirement for HIV/AIDS research is the development of a mucosal vaccine that stimulates humoral and cell-mediated immune responses at systemic and mucosal levels, thereby blocking virus replication at the entry port. Thus, a vaccine prototype based on biologically active HIV-1 Tat protein as antigen and the synthetic lipopeptide, macrophage-activating lipopeptide-2 (MALP-2), asa mucosal adjuvant was developed. Intranasal administration to mice stimulated systemic and mucosal anti-Tat antibody responses, and Tat-specific T cell responses, that were more efficient than those observed after i.p. immunization with Tat plus incomplete Freund's adjuvant. Major linear B cell epitopes mapped within aa 1-20 and 46-60, whereas T cell epitopes were identified within aa 36-50 and 56-70. These epitopes have also been described in vaccinated primates and in HIV-1-infected individuals with better prognosis. Analysis of the anti-Tat IgG isotypes in serum, and the cytokine profile of spleen cells indicated that a dominant Th1 helper response was stimulated by Tat plus MALP-2, as opposed to the Th2 response observed with Tat plus incomplete Freund's adjuvant. Tat-specific IFN-gamma-producing cells were significantly increased only in response to Tat plus MALP-2. These data suggest that Malp-2 may represent an optimal mucosal adjuvant for candidate HIV vaccines based on Tat alone or in combination with other HIV antigens. PMID- 12778474 TI - T cell receptor CDR3 loop length repertoire is determined primarily by features of the V(D)J recombination reaction. AB - The third complementarity-determining region (CDR) of the TCR alpha and beta chains forms loops that engage amino acid residues of peptides complexed with MHC. This interaction is central to the specific discrimination of antigenic peptide-MHC complexes by the TCR. The TCRbeta chain CDR3 loop is encoded by the Dbeta gene segment and flanking portions of the Vbeta and Jbeta gene segments. The joining of these gene segments is imprecise, leading to significant variability in the TCRbeta chain CDR3 loop length and amino acid composition. In marked contrast to other pairing antigen-receptor chains, the TCR beta and alpha chain CDR3 loop size distributions are relatively narrow and closely matched. Thus, pairing of TCR alpha and beta chains with relatively similar CDR3 loop sizes may be important for generating a functional repertoire of alpha beta TCR. Here we show that the TCRbeta chain CDR3 loop size distribution is minimally impacted by TCRbeta chain or alpha beta TCR selection during thymocyte development. Rather, this distribution is determined primarily at the level of variable-region gene assembly, and is critically dependent on unique features of the V(D)J recombination reaction that ensure Dbeta gene segment utilization. PMID- 12778475 TI - CD40-dependent and -independent activation of human tonsil B cells by CpG oligodeoxynucleotides. AB - Immunostimulatory CpG oligodeoxynucleotide (CpG-ODN) sequences are known to directly activate B cells. We investigated the expression of the CpG receptor, Toll-like receptor 9 (TLR9), in human tonsil B cells, and determined functional responses following stimulation by a well-characterized stimulatory CpG containing ODN sequence in the human immune system, ODN 2006. Tonsil B cells were found to express high amounts of TLR9 mRNA and protein, and exposure of B cells to CpG-ODN but not to an inactive control ODN induced a concentration- and time dependent up-regulation of the activation markers CD23, CD25, CD40, CD54, CD80, CD86 and HLA-DR. However, significant induction of proliferation and the release of IL-6, IL-10, IgG and IgM were only noted when B cells were co-incubated with irradiated CD40L-expressing CHO cells. Endogenous IL-10 was identified as a critical mediator of Ig production, whereas all activating effects were independent of IL-6. Further, CpG-ODN counteracted IgE production induced by IL 4. Collectively, these findings suggest a synergistic role of the TLR9/CD40 system and a critical role for the immunomodulatory cytokine IL-10 in the orchestration of CpG-ODN-induced responses in B lymphocytes. PMID- 12778476 TI - The generalized endotoxic principle. AB - Bacterial lipopolysaccharides (endotoxins, LPS) belong to the most potent immunostimulators in mammals. The endotoxic principle of LPS is located in its lipid A moiety, which for Escherichia coli-type LPS consists of a hexaacylated diphosphoryl diglucosamine backbone. This lipid A adopts a cubic inverted aggregate structure from which a conical shape of the molecule can be deduced, whereas the tetraacyl lipid A precursor IVa adopts a cylindrical shape and is endotoxically inactive, but antagonizes active LPS. We hypothesize that non-lipid A amphiphiles with similar physicochemical properties of amphiphilicity, charge, and shape, might mimic the respective lipid A. To test this hypothesis, phospholipid-like amphiphiles with six acyl chains attached to a bisphosphorylated serine-like backbone of varying length replacing the diglucosamine backbone were synthesized. The compound with a short backbone fulfills all criteria of an endotoxic agonist, and that with long backbone fulfills those of an antagonist. This holds true for the human as well as for the murine system. Interestingly, these compounds are inactive in the Limulus amebocyte lysate test which is specific for LPS diglucosamine backbone. These results define a general endotoxic principle and, furthermore, provide new insights into an understanding of early steps of endotoxin action. PMID- 12778478 TI - B cell apoptosis accelerates the onset of murine lupus. AB - To investigate whether the increased rate of lymphocyte apoptosis in systemic lupus erythematosus is involved in the onset of the disease, apoptotic or necrotic T or B lymphocytes from various cell lines were injected intraperitoneally into pre-autoimmune (NZBxNZW)F1 mice (BW) and non-autoimmune BALB/c mice. The intraperitoneal production of cytokines and chemokines, the specific T cell response in the spleen, and the production of anti-histone and anti-dsDNA Ab were investigated. The onset of the disease was characterized by creatinine levels and evaluation of glomerular IgG deposits. In BW, but not in BALB/c mice, injection of apoptotic and not necrotic cells up-regulated IL-6 and IL-10 in resident macrophages. Administration of apoptotic cells augmented the number of Th2 and B lymphocytes recruited in the peritoneal cavity. Only the treatment with apoptotic B cells promoted a systemic Th2 autoimmune response to H2 histones, associated with earlier occurrence of high levels of anti-dsDNA autoantibodies, higher creatinine levels and more numerous glomerular IgG deposits than in BW controls not injected with apoptotic B cells. In genetically susceptible mice exposure to apoptotic of B, but not T, lymphocytes can elicit a Th2 response to H2 histones that helps B cell production of anti-dsDNA Ab and finally triggers the onset of lupus. PMID- 12778477 TI - Epstein-Barr virus (EBV)-positive lymphoproliferations in post-transplant patients show immunoglobulin V gene mutation patterns suggesting interference of EBV with normal B cell differentiation processes. AB - In a model for persistent infection, Epstein-Barr virus (EBV) uses the germinal center (GC) reaction to establish persistence in memory B cells. To study whether EBV adopts to normal B cell differentiation processes also in EBV-associated lymphoproliferative diseases, we micromanipulated EBV(+) cells from biopsies of five patients with post-transplantation lymphoproliferative disease (PTLD) and one unusual Hodgkin lymphoma with many small EBV(+) cells, and analyzed rearranged V genes of single cells. In all cases clonal expansions of EBV(+) B cells were identified. The vast majority of these clones carried mutated V gene rearrangements and a fraction of clones showed ongoing hypermutation. Hence, PTLD likely derive from GC and/or post-GC B cells. In two clones hypermutation occurred in the absence of follicular dendritic and CD4(+) T cells, important interaction partners of normal GC B cells. Furthermore, in one case sustained somatic hypermutation occurred without expression of a functional antigen receptor. Hence, EBV(+) B cells in PTLD can retain or acquire features of GC B cells in an unphysiological setting and may continue to undergo somatic hypermutation uncoupled from normal selection processes, suggesting that EBV interferes with normal B cell differentiation and selection processes in PTLD. PMID- 12778479 TI - Longitudinal analysis of Mycobacterium tuberculosis 19-kDa antigen-specific T cells in patients with pulmonary tuberculosis: association with disease activity and cross-reactivity to a peptide from HIVenv gp120. AB - CD8(+) T cells play a central role in immune protection against infection with Mycobacterium tuberculosis. One of the target epitopes for anti-M. tuberculosis directed CD8(+) T cells is the HLA-A2-restricted 19-kDa lipoprotein peptide VLTDGNPPEV. T cell clones directed against this epitope recognized not only the nominal peptide ligand, but also a closely related peptide (VPTDPNPPEV) from the HIV envelope gp120 (HIV(env) gp120) protein characterized by IFN-gamma release. This cross-reactivity was confirmed in ex vivo in M. tuberculosis 19-kDa tetramer sorted T cells from patients with tuberculosis and in HIVgp120 tetramer-reactive T cells sorted from HIV(+) patients. M. tuberculosis 19-kDa antigen-reactive T cells were present in HLA-A2(+) patients (10/10) with HIV infection with no evidence of M. tuberculosis infection, but they are absent in peripheral blood lymphocytes from healthy HLA-A2(+) individuals (10/10). M. tuberculosis 19-kDa antigen-reactive T cells were elevated in acute pulmonary tuberculosis, declined with response to therapy (7/10 patients) and resided in the terminally differentiated CD8(+) T cell subset. CD8(+) cross-reactive T cells recognizing HIV(env) or M. tuberculosis 19-kDa antigens may contribute to pathogenesis in individuals co-infected with both pathogens and may also present a marker for active tuberculosis. PMID- 12778480 TI - MUC1-derived glycopeptide libraries with improved MHC anchors are strong antigens and prime mouse T cells for proliferative responses to lysates of human breast cancer tissue. AB - Multi-component glycopeptide libraries and single glycopeptides were used for immunization of mice with the aim of inducing strong T helper cell responses to the repetitive sequence of MUC1 expressed by human tumor cells. The glycopeptides and glycopeptide libraries were modeled upon the native human MUC1 amino acid variable number of tandem repeats sequence by introduction of modifications in the MHC anchor positions to optimally fulfil the binding requirements of the A(d) MHC class II molecule in the BALB/c mouse. The immunogenicity of the MUC1 glycopeptides in BALB/c mice was determined by immunization in complete Freund's adjuvant and assaying lymph node T cells for a proliferative response to the glycopeptide used. Strong proliferative responses with stimulation indices over 50 were obtained with anchor-improved glycopeptide libraries as well as with single glycopeptides. Immunization with one of the glycopeptide libraries primed T cells for a proliferative cross-response to the native MUC1 glycopeptide, which by itself was nonimmunogenic. In addition, immunization with the same glycopeptide library primed T cells for a strong response to lysate of a MUC1 expressing human breast cancer, and immunization with the tumor lysate primed T cells for a response to the glycopeptide library. The T cells responding in the assay for proliferation were restricted to the A(d) MHC class II molecule. The results indicate that immunization with MHC anchor-improved MUC1 glycopeptide libraries can effectively prime T helper cells and may induce long-term memory. The approach may be useful in the design of preventive cancer vaccines for use in humans. PMID- 12778481 TI - Rational design of new CpG oligonucleotides that combine B cell activation with high IFN-alpha induction in plasmacytoid dendritic cells. AB - Two different types of CpG motif-containing oligonucleotides (CpG ODN) have been described: CpG-A with high induction of IFN-alpha in plasmacytoid dendritic cells; and CpG-B with little induction of IFN-alpha, but potent activation of B cells. In this study, we demonstrate that CpG-A fail to activate B cells unless plasmacytoid dendritic cells are present. We identified a new set of CpG ODN sequences which induces high levels of IFN-alpha in plasmacytoid dendritic cells but remains capable of directly activating B cells. These new CpG ODN (termed CpG C) are more potent stimulants of B cells than CpG-B due to their ability of directly and indirectly (via plasmacytoid dendritic cells) activating B cells. The sequence of CpG-C combines structural elements of both CpG-A and CpG-B. The most potent sequence, M362, contains a 5'-end 'TCGTCG-motif' and a 'GTCGTT motif', both of which are present in CpG-B (ODN 2006); a palindromic sequence characteristic for CpG-A (ODN 2216); but no poly G motif required for CpG-A. In conclusion, we defined the first CpG-containing sequences that potently activate both TLR9-expressing immune cell subsets in humans, the plasmacytoid dendritic cell and the B cell. CpG-C may allow for improved therapeutic immuno-modulation in vivo. PMID- 12778482 TI - Ganglioside GD3 expression on target cells can modulate NK cell cytotoxicity via siglec-7-dependent and -independent mechanisms. AB - Siglec-7 is a sialic acid binding receptor with inhibitory potential, expressed on human NK cells and monocytes. It has an unusual binding preference for alpha2,8-linked disialic acids, such as those displayed by ganglioside GD3. Here we have investigated whether siglec-7-GD3 interactions are able to modulate NK cell cytotoxicity. Using synthetic polyacrylamide glycoprobes, siglec-7 was found to be masked at the NK cell surface but it could be unmasked by sialidase treatment of NK cells. GD3 synthase-transfected P815 target cells expressed high levels of GD3 and bound strongly to recombinant siglec-7-Fc protein. Surprisingly, GD3 synthase-transfected P815 cells were killed more effectively by untreated cells in a siglec-7-independent manner. However, following sialidase treatment of NK cells, a siglec-7-dependent inhibition of killing was observed. These findings have important implications for NK cell cytotoxicity against tumor cells like melanoma that express high levels of GD3 ganglioside. PMID- 12778483 TI - Proliferating intestinal gamma/delta T cells recirculate rapidly and are a major source of the gamma/delta T cell pool in the peripheral blood. AB - The proliferation, recirculation and repertoire of gut-derived gamma/delta T cells were studied in pigs in vivo. Proliferating gamma/delta T cells (detected by BrdU labeling) are present in all intestinal compartments. In the gut lymph approximately 0.5% of all gamma/delta T cells were proliferating. These gut derived BrdU(+) gamma/delta T cells re-enter the intestinal tissues, and re appear in the intestinal lymph far more often than other cells: about 22% of i.v. injected BrdU(+) gamma/delta T cells were recovered again from the intestinal lymph within 72 h (compare with BrdU(+) B cells 2%, and other BrdU(+) T cells 10%). The contribution of the gut to the migrating gamma/delta T cell pool in the blood became obvious: the proportion of BrdU(+) gamma/delta T cells was three times larger in control versus cannulated pigs. In 9-month-old pigs, clonally expanded T cells were identified in the intestine by complementarity-determining region 3 spectratyping of TCR-delta transcripts. Such expansions were not visible in the blood or intestinal lymph. The distribution of gamma/delta T cells within the intestinal tract is likely to depend to a large degree on the proliferation and the migratory properties of these cells which are different to those of alpha/beta T cells and B lymphocytes. PMID- 12778484 TI - The natural killer cell-mediated killing of autologous dendritic cells is confined to a cell subset expressing CD94/NKG2A, but lacking inhibitory killer Ig like receptors. AB - The cognate NK-DC interaction in inflamed tissues results in NK cell activation and acquisition of cytotoxicity against immature DC (iDC). This may represent a mechanism of DC selection required for the control of downstream adaptive immune responses. Here we show that killing of monocyte-derived iDC is confined to the NK cell subset that expresses CD94/NKG2A, but not killer Ig-like receptors (KIR). Consistent with these data, the expression of HLA-E (i.e. the cellular ligand of CD94/NKG2A) was down-regulated in iDC. On the other hand, HLA-B and HLA-C down regulation in iDC was not sufficient to induce cytotoxicity in NK cells expressing KIR3DL1 or KIR2DL. Remarkably, CD94/NKG2A(+)KIR(-) NK cells were heterogeneous in their ability to kill iDC and an inverse correlation existed between their CD94/NKG2A surface density and the magnitude of their cytolytic activity. It is conceivable that the reduced CD94/NKG2A surface density enables these cells to efficiently sense the decrease of HLA-E surface expression in iDC. Finally, most NK cells that lysed iDC did not kill mature DC that express higher amounts of HLA class I molecules (including HLA-E)as compared with iDC. However, a small NK cell subset was capable of killing not only iDC but also mature DC. PMID- 12778485 TI - Prevention of allergen-specific IgE production and suppression of an established Th2-type response by immunization with DNA encoding hypoallergenic allergen derivatives of Bet v 1, the major birch-pollen allergen. AB - In atopic patients, programming towards a preferential Th2 immunity leads to IgE antibody production and cellular Th2 immunity against otherwise harmless antigens. We report the development of prophylactic and therapeutic DNA vaccines for the major birch-pollen allergen, Bet v 1. We constructed three DNA vaccines, coding for the complete cDNA, coding for two hypoallergenic fragments or coding for a hypoallergenic Bet v 1 mutant. The protective effect was studied in mice pretreated by intradermal DNA injections, then sensitized with Bet v 1 protein. Mice pretreated with any of the three Bet v 1-specific DNA vaccines were protected against allergic sensitization to Bet v 1. Protection was characterized by a lack of Bet v 1-specific IgE production, a lack of basophil activation and an enhanced IFN-gamma expression. DNA vaccines with wild-type Bet v 1 induced strong Bet v 1-specific antibody responses whereas DNA vaccines with hypoallergenic Bet v 1 derivatives induced no (fragments) or only transient (mutant) Bet v 1-specific antibody responses. A therapeutic approach with the fragment-DNA vaccine reduced IgE production and stimulated a sustained Th1 cytokine milieu. Our results demonstrate that DNA vaccines with hypoallergenic forms of the allergen specifically protect against sensitization and suppress established Th2-type responses. This concept may be applied for the development of safe and specific DNA vaccines for the prophylaxis and therapy of allergic diseases. PMID- 12778486 TI - Respiratory syncytial virus-induced CCL5/RANTES contributes to exacerbation of allergic airway inflammation. AB - Severe respiratory syncytial virus (RSV) infection has a significant impact on airway function and may induce or exacerbate the response to a subsequent allergic challenge. In a murine model combining early RSV infection with later cockroach allergen (CRA) challenge, we examined the role of RSV-induced CCL5/RANTES production on allergic airway responses. RSV infection increased CCL5 mRNA and protein levels, peaking at days 8 and 12, respectively. Administration of CCL5 antiserum during days 0-14 of the RSV infection did not significantly alter viral protein expression when compared to mice treated with control serum. In mice receiving the combined RSV-allergen challenge, lungs collected on day 22 exhibited significantly increased numbers of CD4- and CD8-positive T cells. This increase in T cell numbers was not observed in mice receiving alpha-CCL5. On day 43, peribronchial eosinophilia and leukotriene levels were increased in RSV allergen mice. Pretreatment with CCL5 antiserum resulted in decreased recruitment of inflammatory cells to bronchoalveolar and peribronchial regions of the lungs and these reductions were associated with a reduction in both T cell recruitment into the bronchoalveolar space, leukotriene release and chemokine generation. Thus, CCL5 released during RSV infection has a significant effect on the inflammatory response to subsequent allergic airway challenges. PMID- 12778487 TI - Instruction of naive CD4+ T cells by polarized CD4+ T cells within dendritic cell clusters. AB - Cooperation between CD4(+) T cells can enhance the response and modulate the cytokine profile, and defining these parameters has become a major issue for multivalent-vaccine strategies. We explored cooperation using adoptive transfer of two populations of TCR transgenic T cells of different specificity. One was transferred without prior activation, whereas the second was activated for five days by antigen stimulation under polarizing culture conditions. Both populations were transferred into a single adoptive host and then primed by particle-mediated DNA delivery. Polarized Th1 cells (inducers) raised the frequency of IFN-gamma(+) cells within a naive (target) population, whereas Th2 inducers raised the frequency of IL-4(+) and reduced that of IL-2(+) cells. These effects were obtained when the genes for both antigens were on the same particle, favoring presentation by the same dendritic cell, but not when on different particles delivered to different dendritic cells. Autonomy of DC clusters allows linked sets of antigens (e.g. from a single pathogen) to maintain cytokine bias, but allows other independent responses, each with their own set of autonomous clusters. PMID- 12778488 TI - Th2 cells shape the differentiation of developing T cell responses during interactions with dendritic cells in vivo. AB - During priming, naive CD4(+) Th cells differentiate into cells that produce either IFN-gamma or IL-4. Even though the cascade of pathways that induces IL-4 producing Th2 cells has been determined in vitro, the signals promoting Th2 differentiation under physiological conditions remain enigmatic, especially the natural role of the single most important Th2-inducing signal,IL-4. Using Th2 and naive Th cells, each expressing a distinct transgenic TCR, here we show that Th2 cells migrate with the same dynamics as naive Th cells in draining lymph nodes and bind to the same DC, when driven by antigen in complete Freund's adjuvant (CFA). Th2-cell-derived IL-4 deviates CFA-induced Th1 development toward a Th2 phenotype, if both cell populations co-localize in the same T cell area, and are activated simultaneously. Thus, intranodal Th2 cells directly influence Th cell differentiation in vivo, but only under restricted conditions. These findings have implications for the design of cytokine-based therapies and explain the spreading of Th2 responses to multiple aeroallergens in allergic asthma, where naive Th and Th2 cells co-localize in lung-draining lymph nodes. PMID- 12778489 TI - Amino acid composition of alpha1/alpha2 domains and cytoplasmic tail of MHC class I molecules determine their susceptibility to human cytomegalovirus US11-mediated down-regulation. AB - During co-evolution with its host, human cytomegalovirus has acquired multiple defense mechanisms to escape from immune recognition. In this study, we focused on US11, which binds to MHC class I heavy chains and mediates their dislocation to the cytosol and subsequent degradation by proteasomes. To examine which domains of class I heavy chains are involved in this process, we constructed chimeric HLA molecules of US11-sensitive and -insensitive class I molecules (HLA A2 and HLA-G, respectively). Pulse-chase experiments were performed to evaluate protein stability and interactions between class I heavy chains and US11. Flow cytometry was employed to assess the effect of US11 on surface expression of the different chimeras. Our results indicate that the alpha1 and alpha2 domains of HLA molecules are important for the affinity of US11 association. However, the degradation efficiency seems to rely mostly on cytosolic tail residues. We found that the nonclassical HLA-G molecule is insensitive to US11-mediated degradation solely because it lacks essential tail residues. A deletion of the last two tail residues in full-length MHC class I molecules already caused a severe reduction in degradation efficiency. Altogether, our data provide new insights into the mechanism by which US11 down-regulates MHC class I molecules. PMID- 12778490 TI - The immunodominant epitope of lipocalin allergen Bos d 2 is suboptimal for human T cells. AB - We have proposed earlier that the poor capacity of the lipocalin allergen Bos d 2 to stimulate highly allergic subjects' peripheral blood mononuclear cells could be ascribed to endogenous lipocalins and could be related to the allergenic potential of the molecule. Here, we have characterized the proliferative and cytokine responses of human T cell clones against the immunodominant epitope of Bos d 2. We observed, for clone F1-9, that a substitution of aspartic acid for asparagine in the core region of the epitope increased the stimulatory capacity of the peptide about 100-fold in comparison with the natural peptide. For clone K3-2, from a different patient, the substitution of lysine for glutamine or isoleucine for leucine in the core region resulted in about 30-fold and 10-fold increases in the stimulatory capacity of the peptides, respectively. The clones also recognized self-protein-derived peptides but not the peptides derived from other lipocalins. We suggest that the poor recognition of the immunodominant epitope of Bos d 2 can be a factor accounting for Bos d 2-allergic subjects' weak cellular responses. Suboptimal recognition of self and allergen epitopes by T cells may be of significance for the allergenicity of proteins. PMID- 12778491 TI - PU.1 is required for transcriptional activation of the Stat6 response element in the Igepsilon promoter. AB - Signal transducer and activator of transcription 6 (Stat6) has a crucial role in regulation of IL-4-induced gene responses. Stat6-binding sites are present in the promoters of both ubiquitously and cell-type-specifically expressed genes. The promoter regions of IL-4-inducible genes contain cis-acting elements for several transcription factors that act in concert with Stat6 and are also likely to modulate lineage-specific gene expression. We have observed that the Stat6 response element from the B-cell-specific Igepsilon promoter is readily activated upon IL-4 stimulation in B cells but not in non-hematopoietic cells. A minimal low-affinity PU.1-core-binding sequence (5'-AGAA-3') was identified within the Stat6 DNA-binding site in the Igepsilon promoter. Ectopic expression of the myeloid- and B-cell-specific transcription factor PU.1 restored the IL-4 inducibility of the Igepsilon-Stat6 response element in HepG2 cells, and the induction required an intact PU.1-binding sequence. Both the transactivation and the DNA-binding domains of PU.1 were required for induction of Stat6-mediated transcription. The co-operation between PU.1 and Stat6 in transactivation of the Igepsilon gene represents a molecular mechanism for the fine-tuning of cell-type restricted expression of IL-4-induced gene responses. PMID- 12778493 TI - Neutropenia alters lung cytokine production in mice and reduces their susceptibility to pulmonary cryptococcosis. AB - Neutrophils are generally considered to contribute to host defense through their potent microbicidal activity. However, there is accumulating evidence that neutrophils also have an important regulatory role in establishing the balance of Th1 and Th2 responses. This study investigated the role of neutrophils in defense against pulmonary Cryptococcus neoformans infection using neutrophil-depleted BALB/c mice generated by administering mAb RB6-8C5. Neutropenic mice with pulmonary infection survived significantly longer than control mice, but there was no difference between groups infected intravenously. On day 1 of infection, neutropenic mice had significantly smaller fungal burdens than control mice. On day 7, neutropenic mice had significantly higher lung concentrations of IL-10, TNF-alpha, IL-4, and IL-12 than control mice, but there was no difference in IFN gamma and MCP-1 levels. Neutrophils influenced the outcome of cryptococcal infection in mice through mechanisms that did not involve a reduction in early fungal burden. The absence of neutrophils in lung tissue during the initial stages of infection appeared to alter the inflammatory response in a manner that was subsequently beneficial to the host. Higher levels of Th1- and Th2-associated cytokines in neutropenic mice could have simultaneously promoted a strong cellular response while reducing inflammatory damage to the lung. Our results support the emerging concept that neutrophils play an important function in modulating the development of the immune response. PMID- 12778492 TI - Lymphotoxin beta receptor-Ig fusion protein treatment blocks actively induced, but not adoptively transferred, uveitis in Lewis rats. AB - Previous studies have shown that treatment of rodents with a lymphotoxin (LT) beta receptor-Ig fusion protein (LTbetaR-Ig), which binds to both LT and LIGHT, prevents the development of autoimmune diseases, but the mechanism involved is unclear. To explore the potential role of LT or LIGHT in the pathogenesis of autoimmune uveitis, uveitis was induced in Lewis rats either by immunization with an uveitogenic peptide, R16, derived from the interphotoreceptor retinoid-binding protein, or by adoptive transfer of R16-specific T cells. Interestingly, LTbetaR Ig treatment completely prevented actively induced uveitis, but not the adoptively transferred disease. We also show that LTbetaR-Ig-treated R16-injected rats had a significantly decreased T cell response to R16 and that herpesvirus entry mediator (HVEM)-Ig, a fusion protein that blocks LIGHT, also inhibited disease development. Our results suggest that LT or LIGHT plays a critical role in the induction, rather than the effector, phase of the disease. PMID- 12778499 TI - The power to prescribe. PMID- 12778494 TI - Heat shock factor 1-independent activation of dendritic cells by heat shock: implication for the uncoupling of heat-mediated immunoregulation from the heat shock response. AB - The induction of heat shock proteins by heat shock is classically defined as the heat shock response, which is involved in cytoprotection, inflammation and immune responses. Whereas the cytoprotective properties of heat shock have been well characterized, the immunomodulating roles of the heat shock response on the immune system are just emerging. In particular, it is not known whether immunomodulating functions of heat are mediated by the heat shock response. We addressed this question genetically, using a murine model that is unable to mount the heat shock response because of deletion of a major transcriptional factor, heat shock factor 1 (Hsf1). We focused on the roles of heat shock on modulating the functions of dendritic cells (DC) because of their important roles in both innate and adaptive immunity. We found that heat shock matures CD11c(+) DC both in vitro and in vivo, phenotypically and functionally, in the absence of any exogenous inflammatory stimuli. Furthermore, heat-shock-mediated DC maturation is independent of Hsf1, as Hsf1(-/-) DC can be matured by heat shock equally well as wild-type DC. Our novel findings demonstrate that heat shock, one of the most primitive biological responses, can modulate the immune response without the requirement for the transcriptional induction/repression of target genes mediated by Hsf1. PMID- 12778500 TI - Erectile dysfunction: getting to the root of the problem. PMID- 12778501 TI - Effective therapies for managing seasonal allergic rhinitis. PMID- 12778502 TI - Role of vitamins in healthy eating. PMID- 12778503 TI - Addressing the problem of latex glove allergy. PMID- 12778504 TI - Chronic obstructive pulmonary disease: boosting quality of life. PMID- 12778505 TI - Helping to heal the scars of breast cancer treatment. PMID- 12778506 TI - The causes and management of constipation in children. PMID- 12778508 TI - Managing pressure ulcers: the need for pain assessment. PMID- 12778510 TI - A guide to understanding adverse drug reactions. PMID- 12778509 TI - Choosing the right mattress and bed for pressure relief. PMID- 12778511 TI - Pharmacists and nurse prescribers working together to improve patient care. PMID- 12778512 TI - What is nurse prescribing? PMID- 12778513 TI - A new structure for health care in the community. PMID- 12778517 TI - Enteral tube feeding: helping to provide nutritional support. PMID- 12778518 TI - Confirming an asthma diagnosis in the under-fives. PMID- 12778519 TI - A nurse-led initiative to screen and treat hypertension. PMID- 12778520 TI - Switching from an oral regimen to insulin in diabetes. PMID- 12778521 TI - Meeting the health and support needs of older carers. PMID- 12778522 TI - Urinary catheters: reducing the incidence of problems. PMID- 12778523 TI - Postnatal depression: working and learning with mothers. PMID- 12778524 TI - Compression bandaging in leg ulcer management. PMID- 12778525 TI - Paediatric prescribing: the principles and pitfalls. PMID- 12778526 TI - Infected wounds: how to get the diagnosis right. PMID- 12778527 TI - Clinical and financial governance of nurse prescribing. PMID- 12778530 TI - The heavy burden of heart disease. PMID- 12778531 TI - Malaria: how to protect travellers from the big killer. PMID- 12778532 TI - The role of bronchodilators in controlling COPD symptoms. PMID- 12778533 TI - Male catheterisation and the female nurse: still a controversy. PMID- 12778535 TI - Improving men's self-awareness of testicular cancer. PMID- 12778534 TI - Ways forward in promoting teenage sexual health. PMID- 12778536 TI - Simple techniques for managing eczema in children. PMID- 12778537 TI - Using compression hosiery to combat venous leg ulcers. PMID- 12778538 TI - Prescribing HRT: helping women make the right choice. PMID- 12778539 TI - Devising a footcare regimen for people with diabetes. PMID- 12778540 TI - Allaying parental fears over the MMR vaccine scare. PMID- 12778541 TI - Perspective: repression of competition and the evolution of cooperation. AB - Repression of competition within groups joins kin selection as the second major force in the history of life shaping the evolution of cooperation. When opportunities for competition against neighbors are limited within groups, individuals can increase their own success only by enhancing the efficiency and productivity of their group. Thus, characters that repress competition within groups promote cooperation and enhance group success. Leigh first expressed this idea in the context of fair meiosis, in which each chromosome has an equal chance of transmission via gametes. Randomized success means that each part of the genome can increase its own success only by enhancing the total number of progeny and thus increasing the success of the group. Alexander used this insight about repression of competition in fair meiosis to develop his theories for the evolution of human sociality. Alexander argued that human social structures spread when they repress competition within groups and promote successful group against-group competition. Buss introduced a new example with his suggestion that metazoan success depended on repression of competition between cellular lineages. Maynard Smith synthesized different lines of thought on repression of competition. In this paper, I develop simple mathematical models to illustrate the main processes by which repression of competition evolves. With the concepts made clear, I then explain the history of the idea. I finish by summarizing many new developments in this subject and the most promising lines for future study. PMID- 12778542 TI - The additive genetic variance after bottlenecks is affected by the number of loci involved in epistatic interactions. AB - We investigated the role of the number of loci coding for a neutral trait on the release of additive variance for this trait after population bottlenecks. Different bottleneck sizes and durations were tested for various matrices of genotypic values, with initial conditions covering the allele frequency space. We used three different types of matrices. First, we extended Cheverud and Routman's model by defining matrices of "pure" epistasis for three and four independent loci; second, we used genotypic values drawn randomly from uniform, normal, and exponential distributions; and third we used two models of simple metabolic pathways leading to physiological epistasis. For all these matrices of genotypic values except the dominant metabolic pathway, we find that, as the number of loci increases from two to three and four, an increase in the release of additive variance is occurring. The amount of additive variance released for a given set of genotypic values is a function of the inbreeding coefficient, independently of the size and duration of the bottleneck. The level of inbreeding necessary to achieve maximum release in additive variance increases with the number of loci. We find that additive-by-additive epistasis is the type of epistasis most easily converted into additive variance. For a wide range of models, our results show that epistasis, rather than dominance, plays a significant role in the increase of additive variance following bottlenecks. PMID- 12778543 TI - Testing for phylogenetic signal in comparative data: behavioral traits are more labile. AB - The primary rationale for the use of phylogenetically based statistical methods is that phylogenetic signal, the tendency for related species to resemble each other, is ubiquitous. Whether this assertion is true for a given trait in a given lineage is an empirical question, but general tools for detecting and quantifying phylogenetic signal are inadequately developed. We present new methods for continuous-valued characters that can be implemented with either phylogenetically independent contrasts or generalized least-squares models. First, a simple randomization procedure allows one to test the null hypothesis of no pattern of similarity among relatives. The test demonstrates correct Type I error rate at a nominal alpha = 0.05 and good power (0.8) for simulated datasets with 20 or more species. Second, we derive a descriptive statistic, K, which allows valid comparisons of the amount of phylogenetic signal across traits and trees. Third, we provide two biologically motivated branch-length transformations, one based on the Ornstein-Uhlenbeck (OU) model of stabilizing selection, the other based on a new model in which character evolution can accelerate or decelerate (ACDC) in rate (e.g., as may occur during or after an adaptive radiation). Maximum likelihood estimation of the OU (d) and ACDC (g) parameters can serve as tests for phylogenetic signal because an estimate of d or g near zero implies that a phylogeny with little hierarchical structure (a star) offers a good fit to the data. Transformations that improve the fit of a tree to comparative data will increase power to detect phylogenetic signal and may also be preferable for further comparative analyses, such as of correlated character evolution. Application of the methods to data from the literature revealed that, for trees with 20 or more species, 92% of traits exhibited significant phylogenetic signal (randomization test), including behavioral and ecological ones that are thought to be relatively evolutionarily malleable (e.g., highly adaptive) and/or subject to relatively strong environmental (nongenetic) effects or high levels of measurement error. Irrespective of sample size, most traits (but not body size, on average) showed less signal than expected given the topology, branch lengths, and a Brownian motion model of evolution (i.e., K was less than one), which may be attributed to adaptation and/or measurement error in the broad sense (including errors in estimates of phenotypes, branch lengths, and topology). Analysis of variance of log K for all 121 traits (from 35 trees) indicated that behavioral traits exhibit lower signal than body size, morphological, life history, or physiological traits. In addition, physiological traits (corrected for body size) showed less signal than did body size itself. For trees with 20 or more species, the estimated OU (25% of traits) and/or ACDC (40%) transformation parameter differed significantly from both zero and unity, indicating that a hierarchical tree with less (or occasionally more) structure than the original better fit the data and so could be preferred for comparative analyses. PMID- 12778544 TI - Gene flow and the coevolution of parasite range. AB - The geographic range of many parasites is restricted relative to that of their hosts. We study possible evolutionary mechanisms for this observation using a simple model that couples coevolution and demography. The model assumes that the environment consists of two habitats connected by movement and that coevolution is governed by quantitative traits. Our results demonstrate that host gene flow is an important determinant of parasite geographic range. Fluctuations in the rate of host gene flow cause shifts in parasite population densities and associated range expansions or contractions. In extreme cases, changing the rate of host gene flow can lead to global extinction of the parasite. Through a process we term demographic compensation, these shifts in parasite density may occur with little or no change in parasite adaptation to the host. As a consequence, reciprocal adaptation between host and parasite can become uncoupled from the rate of host gene flow. PMID- 12778545 TI - The evolution of virulence in a plant virus. AB - The evolution of virulence is a rapidly growing field of research, but few reports deal with the evolution of virulence in natural populations of parasites. We present here an observational and experimental analysis of the evolution of virulence of the plant virus Cucumber mosaic virus (CMV) during an epidemic on tomato in eastern Spain. Three types of CMV isolates were found that caused in tomato plants either a systemic necrosis (N isolates), stunting and a severe reduction of leaf lamina (Y isolates), or stunting and leaf curl (A isolates). These phenotypes were due to the presence of satellite RNAs (satRNAs) necrogenic (in N isolates) or attenuative (in A isolates) of the symptoms caused by CMV without satRNA (Y isolates). For these three types of isolates, parameters of virulence and transmission were estimated experimentally. For virulence the ranking of isolates was N > Y > A, for transmissibility, Y > A > N. The predictions of theoretical models for the evolution of virulence were analyzed with these parameters and compared with observations from the field. A single infection model predicted adequately the observed long-term evolution of the CMV population to intermediate levels of virulence. A coinfection model that considered competition between isolates with an effect on transmission explained the invasion of the CMV population by N isolates at the beginning of the epidemic, and its predictions also agreed with field data on the long-term evolution of the CMV population. An important conclusion from both models was that the density of the aphid vector's population is a major factor in the evolution of CMV virulence. This may be relevant for the design of control strategies for CMV-induced diseases. PMID- 12778546 TI - Intraspecific competition and mating between fungal strains of the anther smut Microbotryum violaceum from the host plants Silene latifolia and S. dioica. AB - We studied intraspecific competition and assortative mating between strains of the anther smut fungus Microbotryum violaceum from two of its host species, Silene latifolia and S. dioica. Specifically, we investigated whether strains from allopatric host populations have higher competitive ability on their native host species and show positive assortative mating. In general, strains isolated from S. latifolia outcompeted strains isolated from S. dioica on both host species, but in female hosts, heterotypic dikaryons (i.e., dikaryons composed of a haploid strain originating from S. latifolia and a haploid strain originating from S. dioica) were most successful in competition. Furthermore, the latency period was significantly shorter for heterokaryons that contained at least one strain originating from S. latifolia, compared to heterokaryons that only contained strains originating from S. dioica. The frequencies of conjugations between strains originating from S. latifolia were much higher than conjugation frequencies between strains originating from S. dioica. A significant positive correlation was detected between the relative success of strains in competition and in conjugation, suggesting that success of a strain in competition might be partly determined by its swiftness of mating. In addition, reciprocal differences within heterotypic crosses revealed a significant effect of fungal mating type, with mating type a1 being the main determinant of mating pace. The observed differences in infection success, conjugation rate, and latency period in favor of strains from S. latifolia relative to strains from S. dioica on both host species are discussed in an evolutionary context of opportunities for the maintenance of differentiation between different formae speciales upon secondary contact. PMID- 12778547 TI - Test of synergistic interaction between infection and inbreeding in Daphnia magna. AB - It has been proposed that parasitic infections increase selection against inbred genotypes. We tested this hypothesis experimentally using pairs of selfed and outcrossed sibling lines of the freshwater crustacean Daphnia magna, which can be maintained clonally. We studied the performance of selfed relative to outcrossed sibling clones during repeated pairwise clonal competition in the presence and absence of two species of microsporidian parasites. In 13 of the 14 pairs, the selfed clones did worse than the outcrossed ones in the control treatment, but the presence of either parasite did not result in an overall increase in this difference. Rather, it decreased the performance of the selfed relative to the outcrossed sibling in some pairs and increased it in others. Moreover, the two parasite species did not have the same effect in a given pair. This indicates that, contrary to the hypothesis that parasites generally lead to a decreased performance of inbred genotypes, their effect may depend on the genetic background of the host as well as on the parasite species, and suggests that inbreeding can lead to reduced or increased resistance to parasites. Our findings also indicate that there is variation for specific resistance to different species of parasites in the meta-population from which the hosts for this study were obtained. PMID- 12778548 TI - Evidence for strong host clone-parasite species interactions in the Daphnia microparasite system. AB - Organisms are often confronted with multiple enemy species. Defenses against different parasite species may be traded off against each other. However, if resistance is based on (potentially costly) general defense mechanisms, it may be positively correlated among parasites. In an experimental study, we confronted 19 clones from one Daphnia magna population with two bacterial and three microsporidian parasite species. All parasites were isolated from the same pond as the hosts. Host clones were specific in their susceptibility towards different parasite species, and parasite species were host-clone specific in their infectivity, spore production, and virulence, resulting in highly significant host-parasite interactions. Since the Daphnia's resistance to different parasite species showed no obvious correlation, neither general defense mechanisms nor trade-offs in resistance explain our findings. None of the Daphnia clones were resistant to all parasite species, and the average level of resistance was quite similar among clones. This may reflect a cost of defense, so that the cumulative cost of being resistant to all parasite species might be too high. PMID- 12778549 TI - Costs of glandular trichomes in Datura wrightii: a three-year study. AB - Models accounting for genetic variation for resistance to herbivores within plant populations often postulate a balance between the costs of that resistance and its benefits. The production of glandular trichomes by Datura wrightii was shown to be costly in a previous one-year study because plants producing glandular trichomes (sticky plants), a factor conferring resistance to some insect herbivores, also produced 45% fewer seeds than plants producing nonglandular trichomes (velvety plants) when grown in a common garden. Because sticky plants tended to be larger than velvety plants but produced fewer seed capsules, we postulated an allocation trade-off in which velvety plants are more reproduction dominated whereas sticky plants are more growth-dominated. If a greater commitment to vegetative growth eventually allows sticky plants to compensate for reduced seed production, we would expect a reduction or elimination of the cost of resistance over time in this perennial plant. We monitored growth, survival, and seed production of plants from defined crosses of local populations for three years in a common garden when exposed to and protected from herbivores, and with and without supplemental water. The majority of plants exposed to herbivores had died by the end of the study. We used standard life-table methods to determine the net reproductive rate (R0) and the finite rate of increase (lambda) of plants of each trichome type. After three years, when plants were protected from herbivores, sticky plants were 187-245% larger than velvety plants, depending upon irrigation treatment, but sticky plants continued to be less efficient in producing seeds per unit of canopy volume. Even though the total seed production of sticky plants eventually equaled that of velvety plants, the advantage of earlier reproduction by velvety plants increased lambda by 55-230% over that of sticky plants, depending upon herbivore and irrigation treatment. Exposure to herbivores reduced lambda by 69-83%, depending upon plant type and irrigation treatment, whereas supplemental irrigation increased lambda by 29-175%, depending upon plant type and exposure to herbivores. Although there was a large allocation trade-off between growth and reproduction, the benefits of such a trade-off did not emerge before most plants were killed by herbivores. The cost of producing glandular trichomes strictly for herbivore resistance continued to exceed its benefits, and in the absence of other, unmeasured benefits from the suite of life history characters associated with glandular trichome production, natural selection is expected to eliminate this costly resistance trait from D. wrightii populations. PMID- 12778550 TI - Phenotype matching in wild parsnip and parsnip webworms: causes and consequences. AB - According to the geographic mosaic theory of coevolution, selection intensity in interactions varies across a landscape, forming a selection mosaic; interaction traits match at coevolutionary hotspots where selection is reciprocal and mismatch at coldspots where reciprocity is not a factor. Chemical traits play an important role in the interaction between wild parsnip (Pastinaca sativa) and the parsnip webworm (Depressaria pastinacella). Furanocoumarins, produced as plant defenses, are detoxified by the webworms by cytochrome P450 monooxygenases; significant additive genetic variation exists for both furanocoumarin production in the plant and detoxification in the insect, making these traits available for selection. To test the hypothesis that differences in selection intensity affect the distribution of coevolutionary hotspots and coldspots in this interaction, we examined 20 populations of webworms and wild parsnips in Illinois and Wisconsin that varied in size, extent of infestation, proximity to woods (and potential vertebrate predators), and proximity to a chemically distinct alternate host plant, Heracleum lanatum (cow parsnip). Twelve of 20 populations displayed phenotype matching between plant defense and insect detoxification profiles. Of the eight mismatched populations, a logistic regression model related matching probability to two predictors: the presence of the alternate host and average content of xanthotoxin (one of the five furanocoumarins produced by P. sativa). The odds of mismatching were significantly increased by the presence of the alternate host (odds ratio = 15.4) and by increased xanthotoxin content (odds ratio = 6.053). Parsnips growing near cow parsnip displayed chemical phenotypes that were chemically intermediate between cow parsnip and parsnips growing in isolation. Rapid phenotype matching in this system is likely due in part to differential mortality every season; larvae transferred to a plant 30 m or more from the plant on which they developed tended to experience increased mortality over larvae transferred to another umbel on the same plant on which they had developed, and plant populations that mismatched in 2001 displayed a change in chemical phenotype distribution from the previous year. Trait mixing through gene flow is also a likely factor in determining mismatch frequency. Populations from which webworms were eradicated the previous year were all recolonized; in three of seven of these populations, infestation rates exceeded 90%. Our findings, consistent with the geographic mosaic theory, suggest that the presence of a chemically distinct alternate host plant can affect selection intensity in such a way as to reduce the likelihood of reciprocity in the coevolutionary interaction between wild parsnip and the parsnip webworm. PMID- 12778551 TI - Species delimitation and the origin of populations in island representatives of Phylica (Rhamnaceae). AB - Relationships between the closely related island species of Phylica (Rhamnaceae) and a mainland species, P. paniculata, were elucidated using amplified fragment length polymorphisms (AFLPs). Parsimony, neighbor joining, and principal coordinate (PCO) analyses indicated that each of the species studied is distinct. AFLPs were also useful in elucidating the genetic relationships and possible infraspecific origins of different island populations in the Atlantic and Indian Oceans. Phylica nitida on Reunion is likely to have been derived from P. nitida on Mauritius. Although the sampling on New Amsterdam is not extensive, the data are also consistent with the hypothesis that P. arborea on New Amsterdam was derived from a single colonization of P. arborea from Gough Island. Similarly, the Gough Island population appears to have been derived from a single colonization event, but it is so distinct from those on Tristan da Cunha, that there may have been two separate dispersals to Gough and Tristan/Nightingale from different lines of the mainland progenitor. There is also evidence of a recolonization from Gough to Tristan da Cunha. Thus, Phylica arborea is capable of repeated long distance dispersal, up to 8000 km, even though the fruits and seeds are not of a type normally associated with this phenomenon. PMID- 12778552 TI - Interaction between genetic and inductive factors controlling the expression of dispersal and dormancy morphs in dimorphic astigmatic mites. AB - Some astigmatic mites display dimorphic deutonymphs (hypopus) which are facultatively intercalated in their development cycle between protonymph and tritonymph. Such species, among them Glycyphagus privatus and Glycyphagus ornatus show three potential developmental pathways: (1) to bypass the hypopus stage and develop directly from the protonymph to the tritonymph and the subsequent reproductive stage when conditions are favorable; (2) to leave the original site and disperse by means of a phoretic hypopus morph; or (3) to survive inimical life conditions in the natal environment by means of a sedentary hypopus morph. By producing both dispersing (and afterwards at the arrival site reproducing) and sedentary (drought-hardy and dormancy-prone) progeny each single parent attains a selective advantage through a risk-reducing insurance against irregularly fluctuating and often fatal life conditions of their temporary patch habitats. Both genetic heterogeneity and ecological plasticity for hypopus production adapt the Glycyphagus species to cope with variation in the environment. Both traits (for dispersal and survival) are extremely polymorphic with genotypes ranging from low to high propensities for production of each hypopus type. There is a substantial environmental effect on genetic expression such that expression of both morphs depends on the quality of food. This ecological response allows a fast reaction of the mite to the current trophic environment. Phoretic morphs are predominantly expressed at favorable trophic conditions and sedentary morphs at poor trophic conditions. Ecological influences may override genetic propensities and vice versa. Although selection imposed by changing environmental patterns adjusts the frequencies of genotypes over generations and provides for long-term adaptation, the short-term process of environmental induction adapts the population within a generation to transient-habitat disturbances. The interaction of genetic and ecological determinants explains the varying proportions of directly developing mites, phoretic hypopodes, and sedentary hypopodes, in a population at any moment. PMID- 12778553 TI - Seasonal cycles of allozyme-by-chromosomal-inversion gametic disequilibrium in Drosophila subobscura. AB - Allozyme loci are frequently found non randomly associated to the chromosomal inversions in which they are included in Drosophila. Two opposite views compete to explain strong allozyme-by-inversion gametic disequilibria: they result from natural selection or, conversely, merely represent remnants of associations accidentally established at the origin of inversions. Empirical efforts aimed at deciding between adaptive and historical scenarios have focused on the spatial distribution of disequilibria. Yet, the evolutionary significance of these associations remains uncertain. I report here the results of a time-series analysis of the seasonal variation of alleles at six allozyme loci (Acph, Lap, Pept-1, Ao, Mpi, and Xdh) in connection with the O chromosomal polymorphisms of D. subobscura. The findings were: (1) in the segment I of the O chromosome, Lap and Pept-1 allozymes changed seasonally in a cyclical fashion within the ST gene arrangement, but they changed erratically within the 3 + 4 gene configuration; (2) the frequencies of Lap1.11 and Pept-1(0.40) within ST dropped to their lowest values in early and late summer, respectively, when the seasonal level of the ST arrangement is lowest. Furthermore, Lap1.11 and Pept-1(0.40) covary with ST only within these seasons, yet in a fashion inconsistent with these alleles having a major influence on the dynamics of the inversion; (3) seasonal cycling of alleles within inversions were not detected at Acph, Ao, Mpi, and Xdh, yet these loci are nearly monomorphic at the study population, and/or their sampled series were shorter than those for Lap and Pept-1; and (4) simply monitoring allozyme frequencies separately for each inversion proved to be superior, for evidencing the seasonal cycles of the disequilibria, to the use of the D' coefficient of association. Observed seasonal cycles of allozymes within inversions likely reflect natural selection. PMID- 12778554 TI - A comparison of five hybrid zones of the weta Hemideina thoracica (Orthoptera: Anostostomatidae): degree of cytogenetic differentiation fails to predict zone width. AB - Tension zones are maintained by the interaction between selection against hybrids and dispersal of individuals. Investigating multiple hybrid zones within a single species provides the opportunity to examine differences in zone structure on a background of differences in extrinsic factors (e.g., age of the zone, ecology) or intrinsic factors (e.g., chromosomes). The New Zealand tree weta Hemideina thoracica comprises at least eight distinct chromosomal races with diploid numbers ranging from 2n = 11 (XO) to 2n = 23 (XO). Five independent hybrid zones were located that involve races differing from one another by a variety of chromosomal rearrangements. The predicted negative correlation between extent of karyotypic differentiation (measured in terms of both percent of genome and number of rearrangements) and zone width was not found. Conversely, the widest zones were those characterized by two chromosome rearrangements involving up to 35% of the genome. The narrowest zone occurred where the two races differ by a single chromosome rearrangement involving approximately 2% of the genome. The five estimates of chromosomal cline width ranged from 0.5 km to 47 km. A comparative investigation of cline width for both chromosomal and mitochondrial markers revealed a complex pattern of zone characteristics. Three of the five zones in this study showed cline concordance for the nuclear and cytoplasmic markers, and at two of the zones the clines were also coincident. Zones with the widest chromosomal clines had the widest mitochondrial DNA clines. It appears that, even within a single species, the extent of karyotypic differentiation between pairs of races is not a good predictor of the level of disadvantage suffered by hybrids. PMID- 12778555 TI - Size-dependent selection on arrival times in sticklebacks: why small males arrive first. AB - Studies on arrival time to breeding areas show that high-quality males usually arrive first and gain the highest reproductive success. This is generally assumed to be due to phenotype-dependent costs and benefits of early arrival. We show that the opposite arrival order can occur, probably due to selection on poor quality males to increase their chances of reproduction. In a fish species, the threespine stickleback, Gasterosteus aculeatus, small males arrived before larger males at the breeding grounds. Early arrival was costly because predation risk was at its highest at the start of the season and early territory establishment was selected against, as demonstrated by selection coefficients for territory maintenance and hatching success. Large males probably postponed arrival until females were available to decrease predation risk costs and increase offspring production. An experimental study showed that a delay in arrival of large males does not decrease their probability of reproduction, because large males are able to take over nest sites from small males. Small males, on the other hand, are less likely to establish territories in competition with large males but can pay the costs of early arrival in exchange for the benefit of access to territories. Thus, whereas natural selection favors later arrival, sexual selection through competition for breeding territories favors early arrival in small, competitively inferior males. This results in the benefits of early arrival depending on the competitive ability of the male, which favors size-dependent optimal arrival times. PMID- 12778556 TI - Patterns of natural selection on size at metamorphosis in water frogs. AB - Strategies for optimal metamorphosis are key adaptations in organisms with complex life cycles, and the components of the larval growth environment causing variation in this trait are well studied empirically and theoretically. However, when relating these findings to a broader evolutionary or ecological context, usually the following assumptions are made: (1) size at metamorphosis positively relates to future fitness, and (2) the larval growth environment affects fitness mainly through its effect on timing of and size at metamorphosis. These assumptions remain poorly tested, because data on postmetamorphic fitness components are still rare. We created variation in timing of and size at metamorphosis by manipulating larval competition, nonlethal presence of predators, pond drying, and onset of larval development, and measured the consequences for subsequent terrestrial survival and growth in 1564 individually marked water frogs (Rana lessonae and R. esculenta), raised in enclosures in their natural environment. Individuals metamorphosing at a large size had an increased chance of survival during the following terrestrial stage (mean linear selection gradient: 0.09), grew faster and were larger at maturity than individuals metamorphosing at smaller sizes. Late metamorphosing individuals had a lower survival rate (mean linear selection gradient: -0.03) and grew more slowly than early metamorphosing ones. We found these patterns to be consistent over the three years of the study and the two species, and the results did not depend on the nature of the larval growth manipulation. Furthermore, individuals did not compensate for a small size at metamorphosis by enhancing their postmetamorphic growth. Thus, we found simple relationships between larval growth and postmetamorphic fitness components, and support for this frequently made assumption. Our results suggest postmetamorphic selection for fast larval growth and provide a quantitative estimate for the water frog example. PMID- 12778557 TI - Genetic benefits of a female mating preference in gray tree frogs are context dependent. AB - "Good genes" models of sexual selection predict that male courtship displays can advertise genetic quality and that, by mating with males with extreme displays, females can obtain genetic benefits for their offspring. However, because the relative performance of different genotypes can vary across environments, these genetic benefits may depend on the environmental context; in which case, static mating preferences may not be adaptive. To better understand how selection acts on the preference that female gray tree frogs (Hyla versicolor) express for long advertisement calls, I tested for genetic benefits in two realistic natural environments, by comparing the performance of half-sibling offspring sired by males with long versus short calls. Tadpoles from twelve such maternal half sibships were raised in enclosures in their natal pond at two densities. In the low-density treatment, offspring of long-call males were larger at metamorphosis than were offspring of short-call males, whereas in the high-density treatment, offspring of males with long calls tended to metamorphose later than offspring of males with short calls. Thus, although the genes indicated by long calls were advantageous under low-density conditions, they were not beneficial under all conditions, suggesting that a static preference for long calls may not be adaptive in all environments. Such a genotype-by-environment interaction in the genetic consequences of mate choice predicts that when the environment is variable, selection may favor plasticity in female preferences or female selectivity among environments to control the conditions experienced by the offspring. PMID- 12778558 TI - Reproductive character displacement in the acoustic communication system of green tree frogs (Hyla cinerea). AB - Interactions between species can affect the evolution of their sexual signals, receiver selectivity, or both. One commonly expected outcome is reproductive character displacement, whereby adverse consequences of mismating select for greater differentiation of communication systems in areas of sympatry than in areas of allopatry. We found evidence of reproductive character displacement in the acoustic communication system of green tree frogs (Hyla cinerea). The strength of female preferences for the spectral properties of calls that distinguish conspecific calls from those of a closely related congener, H. gratiosa, was greater in areas of sympatry with H. gratiosa than in areas of allopatry. We also found subtle differences in advertisement calls and in the heights of male calling perches when we restricted our comparisons to localities in which H. gratiosa was also breeding (syntopy) with localities where this species was absent. Hyla cinerea and H. gratiosa show only weak genetic incompatibility, but the calls representative of interspecific hybrids were unattractive to females of both parental species. Hybrids might also be at an ecological disadvantages because of different habitat preferences of the two taxa. Thus, selection against production of less fit or less attractive hybrid or backcross offspring are probably the main causes responsible for the differences documented in this paper. PMID- 12778559 TI - The evolution of immune defense and song complexity in birds. AB - There are three main hypotheses that explain how the evolution of parasite virulence could be linked to the evolution of secondary sexual traits, such as bird song. First, as Hamilton and Zuk proposed a role for parasites in sexual selection, female preference for healthy males in heavily parasitized species may result in extravagant trait expression. Second, a reverse causal mechanism may act, if sexual selection affects the coevolutionary dynamics of host-parasite interactions per se by selecting for increased virulence. Third, the immuno suppressive effects of ornamentation by testosterone or limited resources may lead to increased susceptibility to parasites in species with elaborate songs. Assuming a coevolutionary relationship between parasite virulence and host investment in immune defense we used measures of immune function and song complexity to test these hypotheses in a comparative study of passerine birds. Under the first two hypotheses we predicted avian song complexity to be positively related to immune defense among species, whereas this relationship was expected to be negative if immuno-suppression was at work. We found that adult T cell mediated immune response and the relative size of the bursa of Fabricius were independently positively correlated with a measure of song complexity, even when potentially confounding variables were held constant. Nestling T-cell response was not related to song complexity, probably reflecting age-dependent selective pressures on host immune defense. Our results are consistent with the hypotheses that predict a positive relationship between song complexity and immune function, thus indicating a role for parasites in sexual selection. Different components of the immune system may have been independently involved in this process. PMID- 12778560 TI - Genetic and geographic differentiation in the Rio Negro tuco-tuco (Ctenomys rionegrensis): inferring the roles of migration and drift from multiple genetic markers. AB - Among tuco-tucos, Ctenomys rionegrensis is especially amenable to the study of the forces driving population differentiation because of the restricted geographic range it occupies in Uruguay. Within this limited area, the Rio Negro tuco-tuco is limited to sandy soils. It nonetheless exhibits remarkable variation in pelage color, including melanic, agouti, and dark-backed individuals. Two hypotheses have been put forth to explain this pattern: (1) local differentiation and fixation of alternative pelage types by genetic drift under limited gene flow; or (2) fixation by natural selection that may take place even in the presence of gene flow. A previous allozyme study rejected the genetic drift hypothesis on the basis of high inferred levels of migration. New estimates of gene flow from microsatellites and mitochondrial cytochrome b sequences were obtained for C. rionegrensis populations to further test these hypotheses. Much lower levels of gene flow were estimated with these more sensitive markers. Microsatellite-based estimates of gene flow are close to zero and may come closest to estimating current levels of migration. A lack of equilibrium between migration and genetic drift is also strongly suggested by the absence of an isolation-by-distance pattern found in all three genetic datasets. The microsatellite genotype data show that the species is strongly structured geographically, with subpopulations constituting distinct genetic entities. If current levels of gene flow are very low, as indicated by the new data, the local fixation of alternative alleles, including those responsible for pelage color polymorphism, is possible by drift alone. A scenario is thus proposed in which the species expanded in the recent past from a more restricted geographic range and has subsequently differentiated in near isolation, with genetic drift possibly playing a primary role in overall genetic differentiation. The local fixation of pelage color types could also be due to drift, but selection on this trait cannot be ruled out without direct analysis. PMID- 12778561 TI - Viviparity as a constraint on sex-ratio evolution. AB - In polytocous mammals, the sex ratio during gestation can influence a variety of morphological, physiological, and life-history traits because of steroid leakage between fetuses. Similar phenomena have also recently been described for a viviparous lizard. Some of these effects have important fitness consequences by influencing reproductive success later in life. Thus, biasing the sex ratio toward one sex may lead to a decreased fitness for the other sex, and therefore constrain the evolution of skewed sex ratios. By incorporating effects of sex ratio on offspring fitness in a simple sex-allocation model, I show that, under some circumstances (1) skewed sex ratios are predicted to evolve, and (2) this cost can constrain the evolution of skewed sex ratios. PMID- 12778562 TI - Rapid laboratory evolution of adult wing area in Drosophila melanogaster in response to humidity. AB - We examined the evolutionary response of wing area (a trait highly correlated with other measures of body size) to relative humidity (RH), temperature, and their interaction in Drosophila melanogaster, using replicated lines that had been allowed to evolve at low or high humidity at 18 degrees C or at 25 degrees C. We found that after 20 weeks of selection (5-10 generations), low RH lines had significantly greater wing areas than high RH lines in both sexes. This evolutionary response may have resulted from selection of larger flies with a smaller surface area for water loss relative to their weight, or as a correlated response to selection on some other unidentified trait. There were no evolutionary effects of temperature on wing area or cell density. This may have been due to the short duration of the selection experiment, and/or counteracting selection pressures on body size at warm temperature. PMID- 12778563 TI - Effect of high-hydrostatic pressure and temperature on rheological characteristics of glycomacropeptide. AB - The influences of high pressure and temperature on the rheological characteristics of glycomacropeptide (GMP) were studied using a controlled rate rheometer. GMP dispersions at a concentration of 12.5% (w/w) were subjected to high pressure from 100 to 400 MPa for 30 min and temperature from 20 to 80 degrees C for 15 min followed by rheological measurements at a shear rate ranged between 0 and 200 s-1. Shear stress-shear rate data of both pressure and heat induced GMP samples fitted Herschel-Bulkley model well with yield stress. It exhibited shear-thinning behavior with flow behavior index ranged between 0.882 and 0.996. Consistency coefficient and apparent viscosity increased with pressure up to 300 MPa while those parameters decreased at 400 MPa. The rheology of GMP was influenced by temperature. The consistency coefficient and apparent viscosity at 100 s-1 obeyed the Arrhenius relationship with activation energies ranged between 8.17 to 12.38 kJ/mol. Lower activation energy signified lesser molecular aggregation or unfolding of protein molecules during thermal treatment of GMP. PMID- 12778564 TI - Chemistry of buttermilk solid antioxidant activity. AB - Antioxidant activity of buttermilk solids was assessed by analyzing for relative reducing activity, sulfhydryl content, and ferrous and ferric iron binding affinity. These experiments were followed by monitoring the affinity of buttermilk solids to scavenge both hydroxyl and peroxyl radicals in vitro. Notable relative reducing activity of buttermilk solids to L-ascorbic acid (43.80 to 85.85% over a range of 5.0 to 10.0 mg) was attributed in part to the sulfhydryl content (28.8 microM). Buttermilk solids sequestering activity was greater for ferrous than ferric ion. These chemical properties of buttermilk solids corresponded to a significant affinity to scavenge Fenton-induced hydroxyl radical over a range of 5 to 10 mg. A significant affinity of buttermilk solids to protect against lipid peroxidation, tested using an in vitro model lipid system, was also observed at both 0.1 and 0.2% (wt/vol). These findings demonstrated that buttermilk solids possess significant antioxidant activity, thereby suggesting potential use as a value-added ingredient for stabilizing food matrixes against lipid peroxidation reactions. PMID- 12778565 TI - Impaired rennetability of heated milk; study of enzymatic hydrolysis and gelation kinetics. AB - Casein micelles in milk are stable colloidal particles with a stabilizing hairy brush of kappa-casein. During cheese production rennet cleaves kappa-casein into casein macropeptide and para-kappa-casein, thereby destabilizing the casein micelle and resulting in aggregation and gel formation of the micelles. Heat treatment of milk causes impaired clotting properties, which makes heated milk unsuitable for cheese production. In this paper we compared five different techniques, often described in the literature, for their suitability to quantify the enzymatic hydrolysis of kappa-casein. It was found that the technique is crucial for the yield of casein macropeptide and this yield then affects the calculated enzymatic inhibition caused by heat treatment, ranging from 5 to 30%. The technique, which we found to be the most reliable, demonstrates that heat induced calcium phosphate precipitation does not affect the enzymatic cleavage, while whey protein denaturation causes a very slight reduction of enzyme activity. By using diffusing wave spectroscopy, a very sensitive technique to monitor gelation processes, we demonstrated that heat-induced calcium phosphate precipitation does not affect the clotting. Whey protein denaturation does not affect the start of flocculation but has a clear effect on the clotting process. This work adds to a better understanding of the processes causing the impaired clotting properties of heated milk. PMID- 12778566 TI - Gelation mechanism of milk as influenced by temperature and pH; studied by the use of transglutaminase cross-linked casein micelles. AB - Casein micelles in milk are colloidal particles consisting of four different caseins and calcium phosphate, each of which can be exchanged with the serum phase. The distribution of caseins and calcium between the serum and micellar phase is pH and temperature dependent. Furthermore, upon acidification casein micelles lose their colloidal stability and start to aggregate and gel. In this paper, we studied two methods of acid-induced gelation, i.e., 1) acidification of milk at temperatures of 20 to 50 degrees C and 2) decreasing the pH at 20 degrees C to just above the gelation pH and subsequently inducing gelation by increasing the temperature. These two routes are called T-pH and pH-T, respectively. The gelation kinetics and the properties of the final gels obtained are affected by the gelation route applied. The pH-T milks gel at higher pH and lower temperature and the gels formed are stronger and show less susceptibility to syneresis. By using intramicellar cross-linked casein micelles, in which release of serum caseins is prevented, we demonstrated that unheated milk serum caseins play a key role in gelation kinetics and characteristics of the final gels formed. This mechanism is presented in a model and is relevant for optimizing and controlling industrial processes in the dairy industry, such as pasteurization of acidified milk products. PMID- 12778567 TI - Economic feasibility evaluation of microfiltration of milk prior to cheesemaking. AB - A nonlinear programming optimization model was used to evaluate the net revenues and potential profit-ability of microfiltration (MF) prior to cheesemaking in the 3-year period 1998 to 2000, using monthly milk price and composition data. The model identifies the optimal mix of milk resources and determines if MF cheesemaking produces a higher net revenue than conventional cheesemaking that uses NDM and condensed milk for fortification. This study demonstrates the potential of this model to evaluate new technologies in cheese manufacture and improve decision making in the cheese industry. The use of MF produced higher net revenues in 30 out of the 36 mo for both Cheddar and low-moisture, part-skim mozzarella, leading to an appreciable increase in net revenue (vs. conventional cheesemaking) for both cheeses. The benefit from MF in net revenue was greater when the cream price was high. The use of 3X MF yielded the same net revenue as 2X MF. An estimate of manufacturing costs of MF vs. conventional cheesemaking was also made. To this end, the yields of products were calculated by the optimization model, while the production cost of each product was estimated from data of two economic engineering studies and a MF cheesemaking trial. The manufacturing cost of MF Cheddar was slightly higher than the manufacturing cost of conventional Cheddar. However, the benefit in net revenue from the use of MF was estimated to be higher than the difference in manufacturing costs. Moreover, some advantages in the new coproducts of MF Cheddar could outweigh its higher manufacturing cost. The relationships between prices and recoveries of coproducts required to render MF profitable were identified. PMID- 12778568 TI - Effect of temperature of CO2 injection on the pH and freezing point of milks and creams. AB - The objectives of this study were to measure the impact of CO2 injection temperature (0 degree C and 40 degrees C) on the pH and freezing point (FP) of (a) milks with different fat contents (i.e., 0, 15, 30%) and (b) creams with 15% fat but different fat characteristics. Skim milk and unhomogenized creams containing 15 and 30% fat were prepared from the same batch of whole milk and were carbonated at 0 and 40 degrees C in a continuous flow CO2 injection unit (230 ml/min). At 0 degree C, milk fat was mostly solid; at 40 degrees C, milk fat was liquid. At the same total CO2 concentration with CO2 injection at 0 degree C, milk with a higher fat content had a lower pH and FP, while with CO2 injection at 40 degrees C, milks with 0%, 15%, and 30% fat had the same pH. This indicated that less CO2 was dissolved in the fat portion of the milk when the CO2 was injected at 0 degree C than when it was injected at 40 degrees C. Three creams, 15% unhomogenized cream, 15% butter oil emulsion in skim milk, and 15% vegetable oil emulsion in skim milk were also carbonated and analyzed as described above. Vegetable oil was liquid at both 0 and 40 degrees C. At a CO2 injection temperature of 0 degree C, the 15% vegetable oil emulsion had a slightly higher pH than the 15% butter oil emulsion and the 15% unhomogenized cream, indicating that the liquid vegetable oil dissolved more CO2 than the mostly solid milk fat and butter oil. No difference in the pH or FP of the 15% unhomogenized cream and 15% butter oil emulsion was observed when CO2 was injected at 0 degree C, suggesting that homogenization or physical dispersion of milk fat globules did not influence the amount of CO2 dissolved in milk fat at a CO2 injection temperature of 0 degree C. At a CO2 injection temperature of 40 degrees C and at the same total CO2 concentration, the 15% unhomogenized cream, 15% vegetable oil emulsion, and 15% butter oil emulsion had similar pH. At the same total concentration of CO2 in cream, injection of CO2 at low temperature (i.e., < 4 degrees C) may produce a better antimicrobial effect during refrigerated shelf life due to the higher concentration of CO2 in the skim portion of the cream. PMID- 12778569 TI - Serum protein and casein concentration: effect on pH and freezing point of milk with added CO2. AB - The objective of this study was to determine the effect of protein concentration and protein type [i.e., casein (CN) and serum protein (SP)] on pH (0 degree C) and freezing point (FP) of skim milk upon CO2 injection at 0 degree C. CN-free skim milks with increasing SP content (0, 3, and 6%) and skim milks with the same SP content (0.6%) but increasing CN content (2.4, 4.8, and 7.2%) were prepared using a combination of microfiltration and ultrafiltration processes. CO2 was injected into milks at 0 degree C using a continuous flow carbonation unit (230 ml/min). Increasing SP or CN increased milk buffering capacity and protein-bound mineral content. At the same CO2 concentration at 0 degree C, a milk with a higher SP or a higher CN concentration had more resistance to pH change and a greater extent of FP decrease. The buffering capacity provided by an increase of CN was contributed by both the CN itself and the colloidal salts solublized into the serum phase from CN upon carbonation. Skim milks with the same true protein content (3%), one with 2.4% CN plus 0.6% SP and one with 3% SP, were compared. At the same true protein content (3%), increasing the proportion of CN increased milk buffering capacity and protein-bound mineral content. Milk with a higher proportion of CN had more resistance to pH change and a greater extent of FP decrease at the same carbonation level at 0 degree C. Once CO2 was dissolved in the skim portion of a milk, the extent of pH reduction and FP depression depended on protein concentration and protein type (i.e., CN and SP). PMID- 12778570 TI - Sensory threshold of off-flavors caused by proteolysis and lipolysis in milk. AB - The objective of this study was to determine the sensory threshold of off-flavor caused by lipolysis in 2% fat milk and to establish the relationship between increased proteolytic activity in milk and the detection of bitter off-flavor. Homogenized raw milk was held at room temperature for 100 min to allow the native milk lipase to release free fatty acids from the triglycerides. Low and high lipolysis pasteurized milk containing 2% fat were blended together in varying amounts to create a series of six milks with increasing free fatty acid (FFA) concentration for sensory evaluation. Sensory threshold for lipolysis in 2% fat milk was determined by ascending forced-choice procedure, with a series of triangle tests in four sessions with 25 panelists in each session. The group best estimated threshold was the geometric mean of the individual thresholds within each of four panel sessions. The geometric mean best estimated detection thresholds for off-flavors caused by lipolysis in 2% fat milk carried out by native milk lipases were 0.320, 0.322, 0.351, and 0.316 meq of FFA/kg milk for panels 1 to 4, respectively. One third of the panelists detected an off-flavor at or below 0.250 meq of FFA/kg milk. To establish the relationship between proteolysis and detection of off-flavor in pasteurized skim milk, 2800 ppm of CO2 were added to pasteurized skim milk, and it was stored for 27 d at 6 degrees C. Another portion of the same milk was frozen on d 1 at -40 degrees C for use as a low proteolysis portion of the same milk. Decrease in casein as a percentage of true protein (CN/TP) was used as an index of proteolysis. After 27 d at 6 degrees C the milk had a decrease in CN/TP of 4.76% and a standard plate count of 430 cfu/ml. The novel approach of storing milk at 6 degrees C for 27 d with added CO2 blocked microbial growth but allowed proteolytic degradation by milk enzymes to proceed. Before sensory analysis, CO2 was removed by vacuum from the high proteolysis milk and the low proteolysis milk was given the same heat and vacuum. Two triangle tests were performed to determine whether panelists could detect off flavors caused by proteolysis in milk. The threshold detection of off-flavor in skim milk produced by the action of native milk proteases was less than a decrease of CN/TP of 4.76%, but this value is probably near the threshold. PMID- 12778571 TI - Effects of standardization of whole milk with dry milk protein concentrate on the yield and ripening of reduced-fat cheddar cheese. AB - Commercial milk protein concentrate (MPC) was used to standardize whole milk for reduced-fat Cheddar cheesemaking. Four replicate cheesemaking trials of three treatments (control, MPC1, and MPC2) were conducted. The control cheese (CC) was made from standardized milk (casein-to-fat ratio, C/F approximately 1.7) obtained by mixing skim milk and whole milk (WM); MPC1 and MPC2 cheeses were made from standardized milk (C/F approximately 1.8) obtained from mixing WM and MPC, except that commercial mesophilic starter was added at the rate of 1% to the CC and MPC1 and 2% to MPC2 vats. The addition of MPC doubled cheese yields and had insignificant effects on fat recoveries (approximately 94% in MPC1 and MPC2 vs. approximately 92% in CC) but increased significantly total solids recoveries (approximately 63% in CC vs. 63% in MPC1 and MPC2). Although minor differences were noted in the gross composition of the cheeses, both MPC1 and MPC2 cheeses had lower lactose contents (0.25 or 0.32%, respectively) than in CC (0.60%) 7 d post manufacture. Cheeses from all three treatments had approximately 10(9) cfu/g initial starter bacteria count. The nonstarter lactic acid bacteria (NSLAB) grew slowly in MPC1 and MPC2 cheeses during ripening compared to CC, and at the end of 6 mo of ripening, numbers of NSLAB in the CC were 1 to 2 log cycles higher than in MPC1 and MPC2 cheeses. Primary proteolysis, as noted by water-soluble N contents, was markedly slower in MPC1 and MPC2 cheeses compared to CC. The concentrations of total free amino acids were in decreasing order CC > MPC2 > MPC1 cheeses, suggesting slower secondary proteolysis in the MPC cheeses than in CC. Sensory analysis showed that MPC cheeses had lower brothy and bitter scores than CC. Increasing the amount of starter bacteria improved maturity in MPC cheese. PMID- 12778572 TI - Effect of CO2 addition to raw milk on proteolysis and lipolysis at 4 degrees C. AB - Fresh raw milks, with low (3.1 x 10(4) cell/ml) and high (1.1 x 10(6) cells/ml) somatic cell count (SCC), were standardized to 3.25% fat, and from each a preserved (with 0.02% potassium dichromate) and an unpreserved portion were prepared. Subsamples of each portion were carbonated to contain 0 (control, pH 6.9) and 1500 (pH 6.2) ppm added CO2, and HCl acidified to pH 6.2 Milk pH was measured at 4 degrees C. For the preserved low- and high-SCC milks, two additional carbonation levels, 500 (pH 6.5) and 1000 (pH 6.3) ppm, were prepared. Milks were stored at 4 degrees C and analyzed on d 0, 7, 14, and 21 for microbial count, proteolysis, and lipolysis. The addition of 1500 ppm CO2, but not HCl, effectively delayed microbial growth at 4 degrees C. In general, in both the low- and high-SCC unpreserved milks, there was more proteolysis and lipolysis in control and HCl acidified milks than in milk with 1500 ppm added CO2. Higher levels of proteolysis and lipolysis in the unpreserved milks without added CO2 were related to higher bacteria counts in those milks. In preserved low- and high SCC milks, microbial growth was inhibited, and proteolysis and lipolysis were caused by endogenous milk enzymes (e.g., plasmin and lipoprotein lipase). Compared with control, both milk with 1500 ppm added CO2 and milk with HCl acidification had less proteolysis. The effect of carbonation or acidification with HCl on proteolysis in preserved milks was more pronounced in the high SCC milk, probably due to its high endogenous protease activity. Plasmin is an alkaline protease and the reduction in milk pH by added CO2 or HCl explained the reduction in proteolysis. No effect of carbonation or acidification of milk on lipolysis was observed in the preserved low- and high-SCC milks. The CO2 addition to raw milk decreased proteolysis via at least two mechanisms: the reduction of microbial proteases due to a reduced microbial growth and the possible reduction of endogenous protease activity due to a lower milk pH. The effect of CO2 on lipolysis was mostly due to a reduced microbial growth. High-quality raw milk (i.e., low initial bacteria count and low SCC) with 1500 ppm added CO2 can be stored at 4 degrees C for 14 d with minimal proteolysis and lipolysis and with standard plate count < 3 x 10(5) cfu/ml. PMID- 12778573 TI - Microstructure and rheology of yogurt made with cultures differing only in their ability to produce exopolysaccharides. AB - Yogurt was made using an exopolysaccharide-producing strain of Streptococcus thermophilus and its genetic variant that only differed from the mother strain in its inability to produce exopolysaccharides. The microstructure was investigated using confocal scanning laser microscopy, allowing observation of fully hydrated yogurt and the distribution of exopolysaccharide within the protein network. Yogurt made with the exopolysaccharide-producing culture exhibited increased consistency coefficients, but lower flow behavior index, yield stress, viscoelastic moduli and phase angle values than did yogurt made with the culture unable to produce exopolysaccharide. The exopolysaccharides, when present, were found in pores in the gel network separate from the aggregated protein. These effects could be explained by the incompatibility of the exopolysaccharides with the protein aggregates in the milk. Stirring affected the yogurt made with exopolysaccharide differently from yogurt without exopolysaccharide, as it did not exhibit immediate syneresis, although the structural breakdown was increased. The shear-induced microstructure in a yogurt made with exopolysaccharide producing culture was shown to consist of compartmentalized protein aggregates between channels containing exopolysaccharide, hindering syneresis as well as the buildup of structure after stirring. PMID- 12778574 TI - Extraction of immunoglobulin-G from colostral whey by reverse micelles. AB - Separation of immunoglobulin G (IgG) from the other colostral whey proteins was carried out by reversed micellar extraction. The colostral whey was diluted to 5 times its original volume with 50 mM phosphate buffer at pH 6.35 containing 100 mM of sodium chloride. The aqueous solution was then mixed with an equal volume of isooctane containing 50 mM bis-(2-ethylhexyl) sodium sulfosuccinate (AOT), and shaken at 200 rpm and 25 degrees C for 10 min. After extraction, the mixture was separated to the aqueous phase and the reversed micellar phase by centrifugation. This procedure extracted most of the non-IgG proteins to the reversed micellar phase and recovered more than 90% of the IgG in the aqueous phase. The IgG in the aqueous phase had a purity of 90%, and still possessed immunological activity. AOT was not detectable in the aqueous phase. PMID- 12778575 TI - Short-term effect of transition from conventional to automated milking on teat skin and teat end condition. AB - A higher milking frequency, as a consequence of milking with an automated milking system, incorporates a threat to teat condition. To study the effect of transition from conventional to automated milking on teat skin and teat end condition, 40 lactating Holstein-Friesian cows and heifers from a high yielding dairy herd were randomly allocated to either a conventional or an automated milking system group. In the latter group, automated milking was initiated during the study period, while conventional milking was continued in the control group. Teat skin and teat end condition were evaluated weekly on quarter level for all animals from 5 wk before until 8 wk after transition. A high emollient iodine teat dip was used on all cows during the study period. Teat skin condition of the animals in the automated milking system group was consistent from before and during milking with the automated milking system. Rear teats had a better skin and end condition than front teats. Evolution of teat end condition over time between the automated and conventional milking groups was not statistically different. Heifers, however, seemed to be more sensitive to the change than multiparous cows, as their teat end condition slightly decreased. PMID- 12778576 TI - Sources of variation in milk urea nitrogen in Ohio dairy herds. AB - The purpose of this study was to estimate the amount of variation in milk urea nitrogen (MUN) concentrations attributable to test-day, individual cow, and herd effects and to describe factors associated with MUN measurements in Ohio dairy herds. The data came from 24 Holstein herds, half of which were classified as low producing (LP) [rolling herd average (RHA) milk production < 7,258 kg] and half as high producing (HP) herds (RHA production > 10,433 kg). MUN concentration was measured from cow's monthly test-day milk samples. The data were analyzed using multilevel modeling technique in MLwiN, separately for LP and HP herds. The unadjusted mean MUN was 13.9 mg/dl for the HP herds and 11.3 mg/dl for the LP herds. The variance structure was different between the two groups. Most of the variability was found at test-day level in the LP herds, but at herd level in HP herds. MUN was lowest during the first month of lactation, and also season was associated with MUN in both groups. Test-day milk yield, milk fat percentage, and SCC were associated with MUN in the HP herds. With significant explanatory variables in the model, proportionally more of the variation was explained at herd level and less at test day level in both groups. Lower variability in MUN between test days in the HP herds may indicate more consistent day-to-day feeding and management within a herd. The great variability between test days should be considered when interpreting MUN and samples should be collected at the same time of the day to minimize day-to-day variability. PMID- 12778577 TI - Anticipating market effects of new uses for whey and evaluating returns to research and development. AB - As U.S. dairy farms continue to become more productive, increasing demand is a key to improved economic prospects for the dairy industry. One way to expand demand for dairy products is to find new, economically viable uses for milk. Ex ante economic analysis of new uses for agricultural products anticipates the potential market effects of innovations, and provides a basis for evaluating investment in research and development and setting research priorities. This study evaluated potential economic effects of new applications of films and coatings made from whey protein. An economic simulation model was used to predict the likely effects of the innovations on dairy markets. Cost comparisons with existing technologies and interviews with industry officials were the basis for evaluating potential for commercial adoption of the innovations. The economic simulation model traces the projected increased demand for whey through the markets for dairy products and milk. The associated increased demand for milk could result in benefits to U.S. milk producers of $123.0 million in present value terms, compared to a research cost of $ 4.9 million, with the dairy industry, consumers, and taxpayers all contributing. Interpreting the cost of the research program as an investment on behalf of milk producers, the benefits to producers from development of new whey uses represent an annual rate of return between 28 and 33%. These results are useful for evaluating further investment in the whey research program. The methods illustrated here are applicable to the evaluation of a wide range of research and promotion efforts. PMID- 12778578 TI - Effects of once versus twice daily milking throughout lactation on milk yield and milk composition in dairy goats. AB - The effects of once (1X) vs. twice (2X) daily milking throughout lactation on milk yield, milk composition, somatic cell count (SCC), and udder health were studied in 32 Murciano-Granadina dairy goats. Goats were assigned at wk 2 of lactation to two treatment groups; once daily milking at 0900 (1X, n = 17), or twice daily milking at 0900 and 1700 (2X, n = 15). Milk yield was recorded weekly until wk 28, and milk composition and SCC were evaluated for each individual udder half at each milking at wk 2 and 4 of lactation and then, monthly until the end of the experiment. Once daily milking resulted in an 18% reduction in the yield of 4% fat-corrected milk compared to twice daily milking (1.61 vs. 1.95 L/d, respectively). This reduction was more marked from wk 2 to 12 than in mid and late lactation. Response to milking frequency also varied according to parity number where goats of less than four parities suffered more milk yield losses during 1X than older goats. Milk of 1X goats contained higher percentages of total solids (13.6 vs. 12.9%), fat (5.10 vs. 4.62%) and casein (2.57 vs. 2.35%) than milk of 2X goats, but milk protein percentage did not differ between treatments (3.28 vs. 3.20%). Yields of total solids, fat, protein and casein tended to be higher for 2X than 1X. Milk SCC did not differ between treatments. We conclude that application of once daily milking in Murciano-Granadina dairy goats moderately reduced milk yield without negative effects on milk composition and udder health. Losses in milk yield would be reduced if 1X is practiced during mid- or late lactation and in older goats. An increase in labor productivity and a higher farmer's standard of living is also expected. PMID- 12778579 TI - Effects of amounts and degradability of dietary protein on lactation, nitrogen utilization, and excretion in early lactation Holstein cows. AB - Five treatment diets varying in crude protein (CP) and rumen undegradable protein (RUP) were calculated to supply a postruminal lysine to methionine ratio of about 3:1. Diets were fed as a total mixed ration to 65 Holstein cows that were either primiparous (n = 28) or multiparous (n = 37) from 21 to 120 d in milk to determine effects on lactation and nitrogen utilization. Crude protein % and calculated RUP (% of CP) of diets [on a dry matter (DM) basis] were: 1) 19.4, 40 (HPMU), 2) 16.5, 34 (LPLU), 3) 16.8, 40 (LPMU), 4) 16.8, 46 (LPHU), 5) 17.2, 43 (LPHU + UREA), which is the result of adding 0.4% of the diet DM as urea to LPHU. The corn silage-based treatment diets contained an average of 24% acid detergent fiber and 1.6 Mcal/kg net energy of lactation. Milk urea nitrogen (MUN) concentrations and body weights (BW) were used to calculate predicted amounts of urinary nitrogen (N) using the relationship: urinary N (g/d) = 0.0259 x BW (kg) x MUN (mg/dl). Cows fed HPMU had greater CP and RUP intakes, which resulted in higher concentrations of plasma urea nitrogen, rumen ammonia, MUN, and predicted urinary N. Milk yield, fat yield, fat percent, protein yield, and protein percent were not significantly different among treatments. Parity primarily affected parameters that were related to body size and not measurements of N utilization. The interaction of treatment and parity was not significant for any measurements taken. In this study, cows fed LPHU had significantly lower MUN and predicted urinary N without limiting production. These results demonstrate the potential to optimize milk production while minimizing N excretion in lactating dairy cattle. PMID- 12778580 TI - Concentrate feeding strategy in lactating dairy cows: metabolic and endocrine changes with emphasis on leptin. AB - This study aimed to evaluate metabolic and endocrine adaptations to energy intake in multiparous Holstein cows (n = 90; mean 9434 kg energy-corrected milk yield/305 d) over the first 20 wk postpartum and to assess the association of leptin with metabolic, endocrine, and zootechnical traits. Concentrates were fed automatically for 24 h at 30% (C30) or 50% (C50) of total dry matter intake (DMI) from wk 1 to 10 postpartum and at linearly reduced amounts thereafter. Roughage was fed for ad libitum intake. The DMI was measured over 24 h; milk yield and body weight (BW), twice/d; milk composition, 4 times/wk; and milk acetone, weekly. Blood samples for determination of metabolite, hormone, and electrolyte concentrations and enzyme activities were obtained at wk 2 prepartum, and at wk 1 to 16 and at wk 20 postpartum from 0730 to 0900. Body condition scores (BCS) and backfat thickness were measured postpartum and during wk 1, 4, 8, 12, 16, and 20. Energy balance (EB) was considerably lower, but milk yield only slightly lower, in C30 than C50. Metabolic stress was more marked in C30 than C50, expressed by lower, glucose, insulin, insulin-like growth factor-1 (IGF-1), triiodothyronine, milk protein, and lactose concentrations, higher nonesterified fatty acid, beta hydroxybutyrate, growth hormone, and milk acetone concentrations, and an accelerated decrease in BCS and backfat thickness. Nevertheless, C30 adapted successfully and thus maintained high milk yields despite negative EB. Leptin concentrations were lower in C30 than in C50 over the first 20 wk postpartum and were positively associated with BCS, EB, BW, cholesterol, albumin, insulin, and IGF-1; negatively associated with DMI and triiodothyronine; and were higher in cows calving in spring than in fall. Leptin is one among several factors involved in the regulation of energy metabolism and may be important for overall homeostatic and homeorhetic control of metabolism and thus for maintenance of performance. PMID- 12778581 TI - Influence of glyphosate-tolerant (event nk603) and corn rootworm protected (event MON863) corn silage and grain on feed consumption and milk production in Holstein cattle. AB - Two studies were conducted to evaluate the effect of a glyphosate-tolerant (event nk603) and a corn rootworm protected (event MON863) corn hybrid on feed intake and milk production compared with the nontransgenic hybrid and two reference hybrids. In Experiment 1, 16 multiparous Holstein cows were assigned to one of four treatments in replicated 4 x 4 Latin squares with 28-d periods. Diets contained 40% (dry matter [DM] basis) of either 1) glyphosate-tolerant corn silage (GT), 2) nontransgenic control corn silage, or 3) two nontransgenic reference hybrids which are commercially available. Each diet also contained 23% corn grain from the same hybrid that supplied the silage. At ensiling, rapid drying conditions prevailed and the GT hybrid was the last to be harvested which resulted in greater DM content at similar physiological maturity. The 4% fat corrected milk (FCM) yield and DMI were reduced for cows fed the GT corn diet due to the higher DM content of the GT silage (37.1 vs. 33.2 kg/d and 4.05 vs. 3.61% of BW, respectively). There was no effect of the GT diet on milk composition or efficiency of 4% FCM production that averaged 1.43 kg/kg of DM intake for all diets. In Experiment 2, 16 multiparous Holstein cows were assigned to one of four treatments in replicated 4 x 4 Latin squares with 21-d periods. Diets contained 26.7% (DM basis) corn grain from either 1) corn rootworm protected (event MON863) corn hybrid, 2) nontransgenic control corn hybrid, or 3) the same two nongenetically enhanced reference hybrids used in Experiment 1. The 4% FCM yield (34.8 kg/d) and DM intake (4.06% of BW) were unaffected by diet. Efficiency of FCM production (average 1.32 kg/kg of DMI) was not affected by diet. In summary, these two studies indicated that insertion of a gene for glyphosate tolerance or corn rootworm protection into a corn hybrid did not affect its nutritional value (as measured by efficiency of milk production) for lactating dairy cows compared with conventional corn hybrids. PMID- 12778582 TI - Factors affecting application of milk allantoin as an estimator of microbial protein flow to the duodenum under commercial conditions. AB - Three experiments were conducted to determine the effect of diet change, milk sampling technique, and bovine somatotropin (bST) on allantoin output in milk and the use of allantoin as a practical, noninvasive method for estimating microbial protein flow in dairy cattle. In experiment 1, four lactating Holstein cows were used in a 2 x 2 Latin square design with two treatments (ratio of forage to concentrate) and two periods. In experiment 2, six Holstein cows were used in a completely randomized design, and milk was collected by 1) a strip sample collected immediately before milking, 2) a strip sample collected 3 min from start of milking, and 3) a composite sample taken with an autosampler. In experiment three, 10 cows were used in a randomized block design to determine the effect of bST on milk allantoin. Milk samples were taken daily for 21 d, 7 d before, and 14 d after bST administration. In experiment 1, allantoin output (mmol/d) was significantly greater for cows fed the higher ratio of concentrate to forage, and there was a significant change in the amount of allantoin in milk 12 h (first subsequent milking) after a diet change. There was no difference in milk yield or dry matter intake between treatments. In experiment 2, no difference was detected in milk allantoin concentration among the three sampling methods. In experiment 3, milk yield, allantoin concentration, and total allantoin output was significantly increased after bST administration even though dry matter intake (DMI) remained unchanged. During the first 14 d following bST administration, estimates of microbial protein production derived from milk allantoin may be inaccurate due to increased milk production without an increase in DMI. PMID- 12778583 TI - Liver metabolism and production of cows fed increasing amounts of rumen-protected choline during the periparturient period. AB - Forty-eight multiparous Holstein cows were fed treatments consisting of either 0, 45, 60, or 75 g/d of a rumen-protected choline (RPC) source in a completely randomized design from 21 d before expected calving to 63 d postpartum to determine whether choline supplementation to the diet would affect hepatic fatty acid and glucose metabolism, key metabolites in plasma, and cow performance. Dry matter intake (DMI), milk yield, body condition score, and body weights (BW) were similar for cows receiving the four treatments. Feeding RPC tended to increase yields of milk fat, 3.5% fat-corrected milk, and total solids. Plasma concentrations of nonesterified fatty acids and beta-hydroxybutyrate were not different among cows fed the four treatments. Concentrations of triglycerides in liver were similar, but concentrations of glycogen in liver increased as cows consumed increasing amounts of RPC. Hepatic capacity for storage of [1 (14)C]palmitate as esterified products within liver slices tended to decrease as the amount of RPC consumed by cows increased; however, effects of treatment on hepatic capacity for oxidation of [1-(14)C]palmitate to CO2 were not significant. These data imply that choline may increase the rate of very low density lipoprotein synthesis and secretion of esterified lipid products from liver. Hepatic capacities for conversion of [1-(14)C] propionate to CO2 and to glucose in liver were similar among cows fed the four treatments. Collectively, these results suggest that hepatic fatty acid metabolism and cow performance are responsive to increasing the supply of choline during the periparturient period. PMID- 12778584 TI - Performance of lactating dairy cows fed corn as whole plant silage and grain produced from a glyphosate-tolerant hybrid (event NK603). AB - Sixteen multiparous Holstein cows averaging 74 d in milk were used in a replicated 4 x 4 Latin square to compare the effects on animal performance of feeding whole plant silage and grain from a glyphosate-tolerant corn hybrid (event NK603), a nontransgenic control hybrid, and two commercial nontransgenic hybrids (DK647 and RX740). The grain and silage from the four corn hybrids were produced using the same procedures and under similar agronomic conditions at the University of Illinois. On a dry matter (DM) basis, diets contained 30% corn silage and 27.34% corn grain produced either from event NK603, a nontransgenic control, or commercial hybrids. Apart from the DM content of silages, the chemical composition of both grain and silage produced from the four corn hybrids were substantially equivalent. Feeding diets that contained event NK603 and DK647 hybrids tended to decrease DM intake (DMI) compared with the control nontransgenic and RX740. The intakes of crude protein (CP), acid and neutral detergent fiber, and nonfiber carbohydrates were not different for cows fed event NK603 and control diets. The RX740 diet resulted in the highest intakes of fiber and CP, whereas the DK647 diet resulted in the lowest intake of CP. These differences in nutrient intake arose from small variations in both the DMI and the chemical composition of feed ingredients and experimental diets. Production of milk and 3.5% fat-corrected milk; milk fat, CP, and true protein percentage and yield; milk urea N; milk total solids percentage and yield; and somatic cell count were not affected by treatments. These data indicate that the stable insertion of the gene that confers tolerance to glyphosate (event NK603) in the corn line used in this experiment does not affect its chemical composition and nutritional value for lactating dairy cows when compared with conventional corn. PMID- 12778585 TI - Fresh forage and solin supplementation on conjugated linoleic acid levels in plasma and milk. AB - Two experiments were run concurrently to determine the effect of fresh forage consumption on the production and proportions of plasma and milk fat vaccenic acid (VA), conjugated linoleic acid (CLA), and linolenic acid in diary cattle. In experiment 1, the cows consumed 50, 65, and 80% of their feed intake as pasture with the remainder of intake as a barley-based concentrate. The proportion of VA in milk fatty acids increased 12% when pasture intake increased from 50 to 65% of total dry matter intake and VA, CLA, and linolenic acid proportions increased 26, 18, and 27%, respectively, as pasture increased from 65 to 80% of dietary intake. In experiment 2, fresh forage was compared to conserved hay (cut from the same pasture the previous summer) to determine the effect on plasma and milk fat VA, CLA, and linolenic acid. Also, the effect of crushed solin seed (a flax cultivar that is high in linoleic acid) supplementation to the fresh forage diet was determined. Fresh forage compared to conserved hay in the diet, increased the proportion of CLA in the plasma very low density lipoproteins (VLDL) fraction by 71% but had no effect on linolenic acid. Supplementation of the fresh forage diet with a linoleic acid source increased VA and CLA in the plasma VLDL fraction 25 and 58% and slightly decreased the proportion of linolenic acid. Fresh forage, compared to conserved hay, increased milk fat VA and CLA proportions by 22 and 15%. Supplementing the fresh forage diet with linoleic acid from crushed solin seed further increased milk fat VA and CLA proportions 41 and 25%. Solin supplementation in a lactation diet is a superior method to increase CLA levels in milk fat than feeding fresh forage alone. PMID- 12778586 TI - A review of nutritional and physiological factors affecting goat milk lipid synthesis and lipolysis. AB - Although the effect of lactation stage is similar, the responses of milk yield and composition (fat and protein contents) to different types of lipid supplements differ greatly between goats and cows. Milk fat content increases with almost all studied fat supplements in goats but not in cows. However, the response of milk fatty acid (FA) composition is similar, at least for major FA, including conjugated linoleic acid (CLA) in goats and cows supplemented with either protected or unprotected lipid supplements. Goat milk CLA content increases sharply after either vegetable oil supplementation or fresh grass feeding, but does not change markedly when goats receive whole untreated oilseeds. Important interactions are observed between the nature of forages and of oil supplements on trans-10 and trans-11 C18:1 and CLA. Peculiarities of goat milk FA composition and lipolytic system play an important role in the development of either goat flavor (release of branched, medium-chain FA) or rancidity (excessive release of butyric acid). The lipoprotein lipase (LPL) activity, although lower in goat than in cow milk, is more bound to the fat globules and better correlated to spontaneous lipolysis in goat milk. The regulation of spontaneous lipolysis differs widely between goats and cows. Goat milk lipolysis and LPL activity vary considerably and in parallel across goat breeds or genotypes, and are low during early and late lactation, as well as when animals are underfed or receive a diet supplemented with protected or unprotected vegetable oils. This could contribute to decreases in the specific flavor of goat dairy products with diets rich in fat. PMID- 12778587 TI - Models for predicting dry matter intake of Holsteins during the prefresh transition period. AB - The objectives of this study were to develop and validate a model for predicting dry matter intake (DMI) of Holsteins during the prefresh transition period. The original database (ODB) for model development was established by compiling parity, body condition score (BCS), and DMI data during the final 3 wk of gestation from 366 Holsteins fed 24 different diets that were used in eight experiments conducted at three universities. For model validation, a validation database (VDB) was established by compiling data from 333 prefresh transition Holsteins fed 25 different diets that were used in eight experiments conducted at five universities. Dry matter intake during the prefresh transition period was fitted to an exponential function: DMI(t) = a + pe(kt), where DMI(t) = DMI as a percentage of body weight (BW) at time t, a = asymptotic intercept at time- infinity, p = change in intake (kg) from the asymptotic intercept until parturition, k = rate constant influencing the shape of the curve, and t = day relative to parturition expressed as days pregnant--280. The model developed from the ODB predicted DMI of heifers in the VDB with satisfactory accuracy and precision. However, this was not true for cows, probably due to differences in BCS of cows and diets fed to cows from the two data sets. When a subset of cows was selected from each data set that had similar BCS (> 4.0) and were fed similar diets, accuracy and precision of the model predicting DMI was improved. Finally, both databases were combined to develop final models for predicting DMI of heifers and cows. Proposed models for predicting mean daily DMI of heifers and cows during the prefresh transition period were DMI(t) = 1.713-0.688e(0.344t) (R2 = 0.96) and DMI(t) = 1.979-0.756e(0.154t) (R2 = 0.97), respectively. Adjustment factors for animal and dietary factors were generated to demonstrate the plausibility of adaptive fitting of the prediction. The regression coefficients of prediction models (a, p, and k) were affected by BCS and dietary organic macronutrient concentrations. PMID- 12778588 TI - Effects of feeding silage and grain from glyphosate-tolerant or insect-protected corn hybrids on feed intake, ruminal digestion, and milk production in dairy cattle. AB - Lactating dairy cows were used to determine effects of feeding glyphosate tolerant or insect-protected corn hybrids on feed intake, milk production, milk composition, and ruminal digestibility. Corn resistant to European corn borer (Ostrinia nubilalis) infestation (Bt-MON810), or its nontransgenic control (Bt CON), were planted in alternating fields during two successive years. One-half of each strip was harvested for whole plant corn silage and the remainder was allowed to mature and harvested as grain. Effects of feeding diets containing either Bt-MON810 or Bt-CON grain and silage were determined in two experiments (1 and 2) conducted during successive years. In experiment 3, glyphosate-tolerant Roundup Ready corn (RR-GA21) or its nontransgenic control (RR-CON) corn were grown in alternating fields during one cropping season. Diets contained 42 to 60% corn silage and 20 to 34% corn grain from Bt-MON810, RR-GA21, or the appropriate nontransgenic counterpart; treatments were applied using a switchback design. Cows were fed ad libitum and milked twice daily. There were no differences for nutrient composition between silage sources or between grain sources within an experiment. Data for experiments 1 and 2 indicated similar dry matter intake (DMI), 4% fat-corrected milk (FCM) production, and milk composition between Bt MON810 and Bt-CON diets. There were no differences for DMI, 4% FCM production, and milk composition between RR-GA21 and RR-CON diets. There was no difference in ruminal degradability, determined separately for corn silage and corn grain, for RR-GA21 or Bt-MON810-hybrids compared with their respective controls. These data demonstrate equivalence of nutritional value and production efficiency for corn containing Bt-MON810 compared with its control and for RR-GA21 corn compared with its control. PMID- 12778589 TI - Effect of Lactobacillus buchneri, Lactobacillus fermentum, Leuconostoc mesenteroides inoculants, or a chemical additive on the fermentation, aerobic stability, and nutritive value of crimped wheat grains. AB - The preservation of crimped wheat grains by three bacterial inoculants or a chemical additive was compared. Crimped wheat grain [56.8 g dry matter (DM)/kg] was conserved in 1.75-kg plastic bag, mini-silos without treatment, with 4L/tonne of Crimpstore (CS; an additive containing a mixture of ammonium formate, propionate, ethyl benzoate, and benzoate, SAS Kelvin Cave, Ltd., UK) or 1 x 10(5) cfu/g of each of three inoculant additives containing Lactobacillus fermentum (A), Leuconostoc mesenteroides (B), and Lactobacillus buchneri (C). Six replicates were conserved per treatment. Ensiling DM losses, chemical composition, fermentation characteristics, and aerobic stability were measured in the silages after 68 d of ensiling. All the silages were well fermented and remained stable for 84 h after aeration. Subsequently, the rate of deterioration was slowest in crimped grains treated with CS treatment, followed by those treated with inoculant C, while those treated with inoculant A deteriorated most rapidly. Residual water-soluble carbohydrate concentration was higher in crimped grains treated with CS than those treated with the inoculants. Ammonia nitrogen concentrations were lowest in CS-treated crimped grains, followed by inoculants C and A. DM losses were greater in CS-treated crimped grains than in crimped grains treated with inoculants A and C. In vivo digestibility was also measured in Texel cross lambs fed a grass silage basal diet supplemented with the additive-treated crimped grains or a conventional, lamb finisher concentrate. Dry matter intake and digestibility were unaffected by treatment. In conclusion, bacterial inoculants containing L. buchneri are promising preservatives for crimped wheat grains. PMID- 12778590 TI - Intestinal morphology, epithelial cell proliferation, and absorptive capacity in neonatal calves fed milk-born insulin-like growth factor-I or a colostrum extract. AB - Concentrations of nonnutritional factors, such as insulin-like growth factor-I (IGF-I), in bovine colostrum are high and can modulate neonatal gastrointestinal tract development and function. In neonatal calves, we have investigated effects on intestinal epithelial cell morphology, proliferation, and absorption of feeding milk-born human IGF-I (hIGF-I) or a bovine colostrum extract. Calves were fed a milk-based formula containing amounts of nutrients comparable to colostrum for the first 3 d and a milk replacer from d 4 on. Formula and milk replacer contained only traces of nonnutritional factors. In experiment 1, supraphysiological amounts of hIGF-I (3.8 mg/L formula; secreted by transgenic rabbits with their milk) were added to the formula. Xylose appearance in blood (after feeding xylose on d 5) and intestinal traits (after euthanasia on d 8) did not differ between groups. In experiment 2, an extract of first-milked bovine colostrum that provided physiological amounts of IGF-I (0.50, 0.15, and 0.09 mg of IGF-I/L formula on d 1, 2, and 3, respectively, and 0.09 mg of IGF-I/L milk replacer on d 4) was added to formula or milk replacer. Plasma xylose concentration in the control group was transiently higher than in calves fed the colostrum extract. On d 5 (after euthanasia), villus circumferences and heights in small intestine, and epithelial cell proliferation rate in intestine were higher in calves fed the colostrum extract than in controls. In conclusion, orally administered hIGF-I from transgenic rabbits had no effect on the intestinal tract. However, feeding a bovine colostrum extract enhanced intestinal villus size, although it appeared to transiently decrease the absorptive capacity. PMID- 12778591 TI - Genetic parameters of postnatal mortality in Danish Holstein calves. AB - The objective of this study was to estimate genetic parameters of postnatal mortality (PM) in dairy cattle. Data originated from 841,921 Danish Holstein calves. Four binary traits of mortality were considered: D1-14, D15-60, D61-180, and D1-180 with numbers indicating the period of risk in days after birth. The unadjusted frequency of D1-14, D15-60, D61-180, and D1-180 were 0.027, 0.018, 0.020, and 0.066, respectively. A linear sire-model was fitted to the data, and average information-REML was used to estimate (co)variance components. Estimates of direct heritabilities for the four mortality traits ranged from 0.001 to 0.008 but were all significant. D61-180 and D1-180 had the highest direct heritabilities. Maternal heritabilities were very low, ranging from 0.0002 to 0.0015 and significant for D1-14 and D1-180 only. The direct genetic correlation between D1-14 and D15-60, between D15-60 and D61-180, and between D1-14 and D61 180 was 0.73, 0.54, and 0.34, respectively. It indicates that different genes are responsible for early PM (D1-14) and late PM (D61-180). When D61-180 was treated as a different trait for females, males not transferred, and transferred males, the direct heritability was 0.004, 0.008, and 0.034, respectively, but the direct genetic correlations between these three traits were very high. If transfers of calves are getting more common, the importance of including PM in a breeding program will increase, as the genetic variation of PM was considerably higher for transferred calves than for calves that were not transferred. PMID- 12778592 TI - Analysis of vaginal swabs for paternity testing and marker-assisted selection in cattle. AB - Blood is the standard source for DNA analysis, but requires venipuncture of cows by veterinarian and tedious and costly DNA extraction. A procedure was developed for sampling of vaginal cells from cows, establishing a cell lysis protocol using robotics, and applying fluorescent analysis of genetic markers. Two insemination technicians collected vaginal cells from 254 elite Israeli Holstein cows located in 152 herds using commercial Catch-All sample collection brushes. Cells were lysed in a 400-microliters solution, and 5 microliters was used as template for polymerase chain reaction (PCR). Sensitivity of the PCR was enhanced using only 1 microliter of lysed cells. Eight markers of the International Society of Animal Genetics paternity panel were amplified in four separate PCR. ILSTS039, a marker for a quantitative trait loci on BTA14, was amplified in a separate reaction. Genotypes from one to nine genetic markers were obtained for 253 out of 254 samples, and 244 cows had genotypes for at least three markers (96%). Cows that did not inherit either paternal allele for at least two loci were considered not to be daughters of the sire listed. Fifteen cows met this criterion, for a paternity misidentification rate of 6.25%. The frequency of allele 225 of ILSTS039, which was associated with increased milk fat content, was 11.1% in the bull-dam population, similar to the 12% found in the cow population in Israel. The use of vaginal cells for genetic analysis is accurate, as demonstrated by replicated analysis and by comparison to individual and population analysis based on DNA derived from blood. PMID- 12778593 TI - Genotype x environment interaction for milk production in Guernsey cattle. AB - International genetic evaluations that use national genetic evaluation results as input need to acknowledge country boundaries. The current model for international evaluation treats each country as a genetically separate trait, i.e., assumes milk production to be similar within country, but different between countries. The use of cow performance records does not require such restriction, and allows for other statistical models to consider genotype x environment interaction. First-lactation records from 40,000 Guernsey cows in four countries (Australia, Canada, United States, and South Africa) were used to detect and describe genotype x environment interaction for milk production traits. Five statistical models were considered: single-trait across-county (ST), single-trait across country with heterogeneous residual variance (SThet), multiple-trait across country (MT), multiple-trait herd cluster model (HC), and reaction norm model (RR). For the herd cluster model, herds were clustered into groups based on information on herd management, genetic composition, and climate. Reaction norms describe the phenotype expressed by a genotype as a function of the environment, and was modeled by random regression on the herd average for peak milk yield as the descriptor of production environment. Gibbs sampling was used to make inferences about the parameters of interest, and models were compared based on goodness of fit and deviance information criterion. Posterior mode of the heritability for the single-trait model was 0.32, and ranged from 0.15 to 0.53 for models SThet and MT. Posterior mode of the genetic correlations between countries estimated with model MT were generally high (0.78 to 0.90). However, posterior SD were high (up to 0.15 for Australia-South Africa), and values near unity for the genetic correlations were not unlikely. Model HC gave more precise inferences but lower goodness of fit compared with model MT. Results from model RR provided evidence for heterogeneity of genetic variances. This model was least supported by the data, probably because heterogeneity of residual variances was not considered. Among the models in this study, the one with homogeneous genetic and heterogeneous residual variances across countries fitted best to the data, and we expect a model for which the assumption of homogeneous genetic variance is relaxed to show an even better fit to the data. PMID- 12778595 TI - Bayesian estimation of parameters of a structural model for genetic covariances between milk yield in five regions of the United States. AB - Inference about genetic covariance matrices using multiple-trait models is often hindered by lack of information. This leads to imprecise estimates of genetic parameters and of breeding values. Patterns in a genetic covariance matrix can be exploited to reduce the number of parameters and to increase quality of inferences. A structural model for genetic covariances was developed and fitted to milk yield data in five regions of the United States. This was compared with a standard multiple-trait analysis using a deviance information criterion, a measure of quality of fit. Data consisted of 3,465,334 Holstein first-lactation records from daughters of 43,755 sires in five regions of the United States (Midwest, Northeast, Northwest, Southeast, Southwest). Parameters of the structural model included an intercept and effects of measures of genetic and of management similarity on genetic covariances. Genetic similarity depended on the number of records contributed by sires that were common to a pair of regions. Management similarity was a function of the quantity of concentrate used to produce 1000 kg of milk in each pair of regions. The structural and the multiple trait models gave similar estimates of genetic covariances, but the number of parameters was 8 in the former vs. 15 in the latter. Hence, estimates of genetic covariances were more precise with the structural model. A deviance information criterion suggested a slight superiority of the multiple-trait model, although probably within sampling error. For both models, genetic correlations between milk yield in five regions of the United States were larger than 0.93. PMID- 12778594 TI - Quantitative trait loci affecting milk production traits in Finnish Ayrshire dairy cattle. AB - A whole genome scan of Finnish Ayrshire was conducted to map quantitative trait loci (QTL) affecting milk production. The analysis included 12 half-sib families containing a total of 494 bulls in a granddaughter design. The families were genotyped with 150 markers to construct a 2764 cM (Haldane) male linkage map. In this study interval mapping with multiple-marker regression approach was extended to analyse multiple chromosomes simultaneously. The method uses identified QTL on other chromosomes as cofactors to increase mapping power. The existence of multiple QTL on the same linkage group was also analyzed by fitting a two-QTL model to the analysis. Empirical values for chromosome-wise significance thresholds were determined using a permutation test. Two genome-wise significant QTL were identified when chromosomes were analyzed individually, one affecting fat percentage on chromosome (BTA) 14 and another affecting fat yield on BTA12. The cofactor analysis revealed in total 31 genome-wise significant QTL. The result of two-QTL analysis suggests the existence of two QTL for fat percentage on BTA3. In general, most of the identified QTL confirm results from previous studies of Holstein-Friesian cattle. A new QTL for all yield components was identified on BTA12 in Finnish Ayrshire. PMID- 12778596 TI - Calving ease (Co)variance components for a sire-maternal grandsire threshold model. AB - Variance components for a sire-maternal grandsire (MGS) threshold model were estimated from subsets of the US calving ease (CE) database, which includes over 10 million calving records with CE scored 1 (no problem) to 5 (extreme difficulty). Selected records included sire and MGS among the 2601 most frequently appearing bulls. The data were further restricted by requiring at least 20 records in each herd year. Five mutually exclusive sample datasets of approximately 200,000 records each were created based on herd code. The model included random herd-year, sire, MGS, and residual effects and fixed year-season, parity-sex, and birth year of sire and MGS effects. Fewer than 50 iterations were required to reach convergence. The (co)variance component estimates from the five replicates were quite similar. The set of estimates (0.438, herd-year; 0.022, sire; 0.016, MGS; 0.009, sire-MGS) that yielded among the highest heritabilities (0.086, direct; 0.048, maternal) and a correlation of direct and maternal effects near the mean (-0.12) was selected for use in the implementation of a sire-MGS model for CE. PMID- 12778597 TI - Impacts of dairy diagnostic teams on herd performance. AB - This study evaluated impacts of educational diagnostic teams of consultants used to transfer technology to dairy farms. Herd management performance changes were measured by comparing Dairy Herd Improvement data from 38 project farms to data from herds that were geographical contemporaries. The value of focused goals for effecting change was also assessed. Interviews provided producers' perception of project outcomes and insight on organization and conduct of dairy diagnostic teams. Changes observed in project herds were small compared with controls with tendencies for increased herd size and improved milk production per cow. Focused goals had greater impacts on increasing herd size, milk per cow, first lactation peak milk, reducing age at first calving, and percentages of cows with subclinical mastitis. Time, money, facility limitations, labor, and alternative priorities were the most cited constraints to implementing changes. Satisfaction scores of producers were significantly related to the degree that team recommendations were followed. Improved attitudes, quality of life, and financial well-being were benefits listed by a majority of producers from participation in the project. If similar projects were to be offered, 83% said they would participate again, and 69% indicated they would pay at least some of the costs. Project farms served as demonstration farms for 1930 other producers in their respective locales, resulting in a multiplier effect of original advice given by consultant teams. Suggestions by farmer participants for improvements in dairy diagnostic teams included needs for at least some unbiased team members, more frequent meetings, more follow-up on recommendations, and consistency of recommendations with family goals. PMID- 12778598 TI - Modification of the Penn State forage and total mixed ration particle separator and the effects of moisture content on its measurements. AB - The Penn State Particle Separator has led to widespread measurement of forage and total mixed ration (TMR) particle size. However, a large proportion of small particles may pass through both sieves when a TMR is analyzed, and field research has suggested that both shaking frequency and sample dry matter may affect the results. The objectives of this project were to test the effects of an additional sieve with a smaller aperture size, shaking frequency, and sample moisture content on results obtained. A sieve was constructed out of wire with a nominal size aperture of 1.18 mm. Samples of alfalfa haylage, corn silage, and a TMR were shaken at frequencies of 0.9, 1.1, and 1.6 Hz with a 17-cm stroke length. Reducing shaking frequency to 0.9 Hz resulted in more material being retained on the 19.0-mm sieve for all sample types, increasing the geometric mean. Increasing frequency to 1.6 Hz did not affect the geometric mean, but did result in a greater amount of corn silage falling through the 1.18-mm sieve. For alfalfa haylage, moisture content between 57.4 and 35.6% did not affect results; however, for corn silage, less moisture increased the percentage of particles less than 1.18 mm and decreased the geometric mean. For both sample types, further drying caused a greater proportion of small particles and a smaller geometric mean. We suggest using a third sieve and shaking at 1.1 Hz or greater with a stroke length of 17 cm when using the Penn State Particle Separator to analyze forage particle size. PMID- 12778600 TI - Central auditory processing disorder (CAPD). PMID- 12778599 TI - ACAAI conducts seventh annual Nationwide Asthma Screening Program. PMID- 12778601 TI - The Mediterranean food guide pyramid. PMID- 12778602 TI - Poison ivy 'leaves of three, let it be!'. PMID- 12778603 TI - The complex role of the school nurse. Promoting who we are and what we do. PMID- 12778604 TI - Trichotillomania. PMID- 12778605 TI - "You hit your head?" After mild traumatic brain injury: helping school staff meet the needs of students and families. PMID- 12778606 TI - Dental dilemma. Should schools provide on-site oral care for children? PMID- 12778607 TI - SARS. Nurses among first quarantined. PMID- 12778608 TI - Delving into bone density. PMID- 12778609 TI - Severe acute respiratory syndrome. The Mount Sinai experience. PMID- 12778610 TI - The Dr. Peter Centre. Harm reduction nursing. PMID- 12778611 TI - Recruitment study. Why students choose nursing. PMID- 12778612 TI - Survey of CKD patients reveals significant gaps in knowledge about kidney disease. Part 2. PMID- 12778613 TI - Connecting the dots: caring for the patient with progressive CKD. AB - The approach we take to CKD in the KDIPPP Clinic represents a complicated regimen. It's not something we can do by ourselves, and it requires an integrated team approach. The team approach includes many different individuals all working together (see Fig. 2, p. 25). If we are to be successful, the initiative must include collaboration from the primary care physicians (PCPs), providers, and patients. Patients need to be screened yearly for serum creatinine, blood glucose, and blood pressure if they are at risk for CKD. Creatinine should be expressed as a GFR. Examine the urine to see if it is abnormal and check for microalbuminuria. The PCP, the patient, and the nephrologist must team up to establish the balance of care (see Fig. 3, p. 26). As the CKD patient progresses from the presence of risk factors to CKD Stages 1, 2, 3, and finally 4 and 5, the input from the PCP and the nephrologist gradually changes, from nearly full-time PCP care in the early stages to nearly full-time nephrology team care in the later stages. When patients migrate through progression, prevention, education, access placement, and RRT, the degree of involvement and interaction must be individualized. I recommend follow-up every year or two for patients in CKD Stages 1 and 2; follow-up every six months for Stage 3, and increased nephrology input as CKD progresses. We need to work together to treat these patients. If we do, then we're going to end up with better patient outcomes and better lives. The KDIPPP Clinic has been successful at meeting these goals. PMID- 12778614 TI - Automating patient records. An independent facility reaches for excellence. PMID- 12778615 TI - Can we find the road to improvement? PMID- 12778617 TI - Investing in the lifeline: the value of a vascular access coordinator. PMID- 12778616 TI - Vascular access use and outcomes in the U.S., Europe, and Japan: results from the Dialysis Outcomes and Practice Patterns Study. PMID- 12778618 TI - Making cents of logistics. 15 ways to improve inventory control. PMID- 12778619 TI - The patients and staff who try our patience. PMID- 12778620 TI - Running water: designing the dialysis clinic water room. Part 1. PMID- 12778621 TI - Reuse: is it right for your facility? PMID- 12778622 TI - Oral pathology. PMID- 12778623 TI - The use of bovine porous bone mineral in combination with collagen membrane or autologous fibrinogen/fibronectin system for ridge preservation following tooth extraction. AB - Preservation of the alveolar ridge following tooth extraction is desirable since it facilitates placement of endosseous implants and may improve the adverse esthetics often associated with fixed partial dentures. The purpose of this study was to compare the clinical effectiveness of bovine porous bone mineral (BPBM) used as a graft material combined with either guided tissue regeneration (GTR) or with the autologous fibrinogen/fibronectin system (AFFS) in preserving alveolar ridges following tooth extraction. Twenty-six patients who required extraction of two or more anterior or bicuspid teeth participated in a split-mouth design study. Following tooth extraction and elevation of a buccal full thickness flap, sockets were filled with bovine porous bone mineral which was then covered with either a collagen membrane or mixed and covered with an AFFS system. An acrylic stent served as a reference point for measurements. Primary flap closure was achieved in all surgical sites, and reentry surgery was performed at 6 months. Reentry surgery showed that BPBM/GTR sites presented with [1] significantly more internal socket bone fill (6.04 +/- 0.21 mm vs. 4.98 +/- 0.26 mm), [2] less, although not statistically significant, resorption of alveolar bone height (0.23 +/- 0.28 mm vs. 0.3 +/- 0.21 mm), and [3] significantly less horizontal resorption of the alveolar bony ridge as compared to BPBM/AFFS (1.06 +/- 0.28 mm vs. 2.60 +/- 0.25 mm). This study suggests that treatment of extraction sockets with a combination of bovine porous bone mineral and guided tissue regeneration is of slightly more benefit in preserving alveolar ridge dimensions following tooth extraction than treatment with a combination of bovine porous bone mineral and the autologous fibrinogen/fibronectin system. PMID- 12778624 TI - [Surgical treatment of ascending aortic aneurysm]. AB - Despite of its low incidence ascending aortic aneurysm is an extremely important disease due to its continuous life threatening feature. In spite of advanced diagnostic and surgical and anesthetic methods--developed in the last decade- aneurysm operations have high mortality rate particularly in cases of complications such as rupture or dissection (30-45%). Author discusses data of 55 patients operated on from 1994 to 2001. Overall mortality was 16.4% (28% for acute and 6.6% for scheduled cases) which is considered to be good in comparison to the literature. In good results contribute introduction of new surgical techniques, use of tissue glue, and hypothermic circulatory arrest. Author summarises symptoms, diagnostic modalities, surgical indication, methods of operation, early and late complications based on data from international literature and on his own experience. PMID- 12778625 TI - [Enzyme replacement therapy in Gaucher disease: monitoring visceral and bone changes with MRI]. AB - INTRODUCTION: Gaucher disease (GD) is the most common lysosomal storage disorder. It can be successfully treated with infusion of the modified deficient enzyme. AIM: Visceral and skeletal changes during enzyme replacement therapy (ERT) in patients with type 1 Gaucher disease (GD) were studied by radiological methods. METHODS: Eight patient with GD (8-39 years) were examined, who underwent ERT for 1-8 years (30-80 IU/bwkg/months Ceredase or Cerezyme). MRI from the upper abdomen were made before ERT was started and once a year during ERT, that allowed measurement of hepatic and splenic volume. The bone changes were depicted by MRI and conventional X-ray images. Bone marrow infiltration was estimated by MRI scans of femora with semiquantitative method. RESULTS: Hepato- and splenomegaly were detected before ERT, which decreased in each patient during therapy. Decrease of liver volume was more expressed in patients with splenectomy. The infiltration of the bone marrow with Gaucher cells has reduced during treatment in six cases. Decreases of hepatosplenomegaly occured earlier and faster, than the improvement of bone disorders. Two splenectomised patients exhibited 4 episodes of bone crisis diagnosed by MRI. Their symptoms regrediated on increased enzyme doses. The grade of bone marrow infiltration and concurrent bone complications could not be detected by conventional X-ray films. CONCLUSIONS: We conclude that the use of ERT in all patients leads to marked improvements in visceral and skeletal pathology of patients with Gaucher disease. The volume of the liver and the spleen, and the extension of bone marrow infiltration can be measured precisely by MRI, so it can be the method of choice in the diagnosis and in the monitoring of response to ERT. MRI helps to determine the appropriate enzyme dosage and to establish the necessity of elevation the enzyme dose. MRI can be practicable in the differential diagnosis of bone crisis and osteomyelitis. PMID- 12778626 TI - [Triple helix: a new promise for gene therapy]. AB - AIM: This publication focuses on the inhibition of gene expression by triplex forming oligonucleotides, which is a promising approach to the treatment of dominant genetic diseases. The most common cause for autosomal dominant retinitis pigmentosa is a mutation in the rhodopsin gene. METHODS: Two psoralen-linked triplex-forming oligonucleotides were used to inhibit the expression from a plasmid carrying the rhodopsin and green fluorescent protein fusion gene. Following in vitro triplex formation, UVA irradiation was used to activate the psoralen moiety to form covalent photoadducts between the three strands. The samples where then transfected into human fibrosarcoma cells and analysed for green fluorescence. RESULTS: Photoadducted triple helix formation resulted in reduction of gene expression by as much as 90% and this effect persisted for at least 72 hours. However, expression levels from a cotransfected control plasmid were unaffected. Mutations at one of the triplex binding sites within the rhodopsin gene also abolished the effect of the corresponding triplex forming oligonucleotide, without diminishing the inhibition by the other oligo. Northern blots indicated that photoadducted triplex formation blocked the progression of the RNA polymerase, resulting in truncated transcripts. CONCLUSION: The authors conclude, that psoralen linked triplex forming oligonucleotides are efficient and specific tools for blocking gene expression from the human rhodopsin gene. PMID- 12778627 TI - [Pleiotropic effect of the micronized fenofibrate: the reduction of plasma chlamydia pneumoniae antibody levels in patients with coronary artery disease]. AB - INTRODUCTION: A growing amount of data suggest that atherosclerosis is an inflammatory disease and in it's development Chlamydia pneumoniae infection may contribute. Recent studies have shown that administration of micronized fenofibrate reduces the plasma levels of several markers of the inflammatory response. AIM: The aim of the authors was to evaluate the effect of micronized fenofibrate on the lipids and Chlamydia pneumoniae antibody levels of 20 patients with coronary artery disease. METHODS: The plasma total cholesterol, HDL cholesterol, triglyceride, lipoprotein (a), ApoA1, ApoB, fibrinogen and Chlamydia Ig A, IgG and IgM antibody concentrations were examined. The patients were on strict lipid lowering diet and were treated by daily 200 mg micronized fenofibrate for 3 weeks. RESULTS: A significant reduction of the total cholesterol (18.3%), LDL cholesterol (17.7%), triglyceride (37.8%), ApoB (18.4%), fibrinogen (16.1%) levels was observed. The concentration of HDL cholesterol (17.0%) and ApoA1 (12.2%) showed a significant elevation. The Chlamydia pneumoniae IgM antibody (characterizing the acute infections) was not detectable. The IgA antibody level decreased from 13.0 EIU to 12.3 EIU (p < 0.05) and IgG from 85.1 EIU to 78.7 EIU (p < 0.05). CONCLUSIONS: Beside the expected lipid lowering effect of the micronized fenofibrate a significant reduction in the plasma Chlamydia pneumoniae IgA and IgG antibody levels was observed supporting the anti-inflammatory, pleiotropic effect of this drug. PMID- 12778628 TI - [Thoughts about the treatment of tumors based on the work of Professor Tibor Verebely]. PMID- 12778629 TI - [Klinikai Fuzetek--100 years ago]. PMID- 12778630 TI - 401(k) plan asset allocation, account balances, and loan activity in 2001. PMID- 12778631 TI - Retiree health benefits: savings needed to fund health care in retirement. PMID- 12778632 TI - Unannounced JCAHO surveys mean more planning, not less. PMID- 12778633 TI - Planning more important for unannounced surveys. PMID- 12778634 TI - NC hospital issues quality report cards. PMID- 12778636 TI - Closed-circuit TV wins fans in children's hospital. PMID- 12778635 TI - Patient Satisfaction Planner. 'Making it personal' improves patient care. PMID- 12778637 TI - Study shows QI helps heart attack patients. PMID- 12778638 TI - 'Brewing cataclysm' in emergency response? PMID- 12778639 TI - Reduce infections with root-cause analysis. PMID- 12778644 TI - Easing menopausal symptoms with dietary phytoestrogens. PMID- 12778645 TI - Helping coeliac disease patients adapt to a gluten-free diet. PMID- 12778646 TI - Urinary catheterisation in cancer: improving quality of life. PMID- 12778647 TI - Why promoting good bowel health may help prevent cancer. PMID- 12778648 TI - The nurse's contribution to the care of dermatology patients. PMID- 12778650 TI - Prescribing pitfalls: cyproterone acetate with ethinyloestradiol. PMID- 12778649 TI - Interpreting and analysing practice prescribing data. PMID- 12778651 TI - Selecting an appropriate treatment for infected wounds. 2. PMID- 12778653 TI - Wound contact layers under compression. PMID- 12778654 TI - Online nutrition resources. PMID- 12778655 TI - Controlling blood pressure with a healthy lifestyle. PMID- 12778656 TI - Type 2 diabetes. Adjusting to the diagnosis. PMID- 12778657 TI - Exercise myths and facts. PMID- 12778658 TI - Beyond broccoli. PMID- 12778659 TI - Vegetarian and vegan meal planning. PMID- 12778660 TI - Disease, treatment and oral health. PMID- 12778661 TI - Diabetes quiz. How much do you know about food safety? PMID- 12778662 TI - Supermarket smarts. Jams, jellies, and syrups. PMID- 12778663 TI - Treating heart failure. PMID- 12778664 TI - For parents. When divorce enters the picture. PMID- 12778665 TI - The physician-dentist liaison. PMID- 12778666 TI - Advanced technology brings equine dentistry into light of day. PMID- 12778667 TI - CDC release "refreshes" Hispanic parents' knowledge of good oral health habits. PMID- 12778668 TI - Oral health and the pregnant patient. PMID- 12778669 TI - The facts about cold sores. PMID- 12778670 TI - New hand hygiene guidelines from the CDC. PMID- 12778671 TI - Hygiene recall: is it a waste of time? PMID- 12778672 TI - The contact lens porcelain veneer. PMID- 12778674 TI - Simplified cosmetic denture fabrication utilizing the patient's old denture. PMID- 12778673 TI - Maximizing aesthetics using a combined periodontal-restorative protocol. PMID- 12778675 TI - Making a difference by making someone's smile. PMID- 12778676 TI - Optimum dental care, Part 3. Case finishing. PMID- 12778677 TI - Everything I know about endodontics, I learned after dental school, Part 1. PMID- 12778678 TI - Predictable endodontic success. The apical control zone. PMID- 12778679 TI - Obturation of the root canal system. PMID- 12778680 TI - The future of periodontal diagnostic testing. PMID- 12778681 TI - The evolution of dentin bonding. PMID- 12778682 TI - Clinical applications of appliance therapy in the general practice, Part 1. Interceptive orthodontics in children. PMID- 12778683 TI - How staff can tap a gold mine of production with just a nugget of problem-solving skills. PMID- 12778684 TI - Managing your practice through team accountability. PMID- 12778685 TI - Ethics in action. PMID- 12778686 TI - Improve your suctioning technique. PMID- 12778687 TI - Tracheal suction catheters. PMID- 12778688 TI - Is your pediatric patient a victim of bullying? PMID- 12778689 TI - Pairing quick-hire tactics with retention strategies. PMID- 12778690 TI - Thalidomide makes a comeback. PMID- 12778691 TI - Inflammatory bowel disease. PMID- 12778692 TI - Tips for teaching the elderly. PMID- 12778693 TI - IABP therapy. Getting patients treatment fast. PMID- 12778694 TI - Does error + injury = negligence? PMID- 12778695 TI - Metallo-enzyme catalysis. AB - All organisms depend upon metallo-enzymes. The dependence arises from the inability of individual organic side-chains of proteins to activate molecules such as H2, N2, CH4 and CO and their weakness in hydrolysing many simple compounds such as many peptides, phosphates, even urea. The metal ion sites have been found to be 'designed' for selective uptake and catalytic activity. In this article a few examples will be used to illustrate these points. For more details of all the examples see the reference at the end of this article to Messerschmidt et al. (2001). PMID- 12778696 TI - Automated carbohydrate synthesis to drive chemical glycomics. AB - This feature article describes the development of the first automated solid-phase oligosaccharide synthesizer. A series of chemical challenges had to be addressed to accomplish this breakthrough and provide rapid access to oligosaccharides of biological significance. Accelerated synthesis of glycoconjugates promises to greatly impact the emerging field of glycobiology. Chemical glycomics uses synthetic carbohydrates and analogs to study their role in recognition, signal transduction pathways and other events of fundamental biomedical significance and shapes up to become the next major wave in biomedical research. The automated synthesis of a novel malaria vaccine candidate is discussed to illustrate the medical potential of chemical glycomics. PMID- 12778698 TI - Self-assembled dimers with supramolecular chirality. AB - The possibilities to form dimeric capsules with supramolecular chirality from tetraurea calix[4]arenes composed of two different phenolic urea units are discussed in general and demonstrated experimentally for various selected examples. PMID- 12778697 TI - Bioinspired synthesis of new silica structures. AB - Silicon and oxygen are the two most abundant elements in the Earth's crust but despite the vast scientific literature on crystalline and amorphous silica, new chemistries, structures and applications continue to be discovered for compounds formed from these elements--thus we present here for the first time the formation of new amorphous silica structures that were uniquely synthesized by a bioinspired synthetic system. PMID- 12778699 TI - Bimetallic nanocatalysts for the conversion of muconic acid to adipic acid. AB - Adipic acid (2) production currently entails use and generation of environmentally harmful materials: an efficient catalyst, consisting of nanoparticles of Ru10Pt2 anchored within the pores of mesoporous silica, facilitates the production of (2) by hydrogenating muconic acid, that may be derived biocatalytically from D-glucose. PMID- 12778700 TI - Unwinding of DNA polymerases by the antitumor drug, cis diamminedichloroplatinum(II). AB - The helix-turn-helix motifs of the DNA binding domains of human polymerase-alpha and polymerase-kappa are dramatically perturbed upon binding to cisplatin with concomitant release of zinc. PMID- 12778701 TI - First comparative emission assay of single-wall carbon nanotubes--solutions and dispersions. AB - A variety of single-wall carbon nanotube samples were tested in steady-state and time-resolved emission experiments, where it was found that chemical functionalization and different degrees of oxidation affect the emission behavior. PMID- 12778702 TI - Dicationic [(BINAP)Pd(solvent)2]2+[TfO-]2: enantioselective hydroamination catalyst for alkenoyl-N-oxazolidinones. AB - Dicationic (BINAP)palladium(II) complex induced high enantioselectiviies in the addition of primary (and secondary) aromatic amines to alpha,beta-unsaturated oxazolidinones (up to 93% ee). PMID- 12778703 TI - Sensitised near-infrared emission from lanthanides using a covalently-attached Pt(II) fragment as an antenna group. AB - In a series of heterodinuclear complexes in which a Pt(PPh3)2(catecholate) chromophore is covalently linked to a lanthanide tris(diketonate) unit, sensitised near-IR emission from Yb(III), Nd(III) and Er(III) occurs on excitation of the Pt(II) chromophore at 520 nm. PMID- 12778704 TI - The truly symmetrical doubly-bridging mode of trimethylphosphine in a dinuclear rhodium complex. AB - The dinuclear complex [Rh2I2(mu-CPh2)2(mu-PMe3)] was prepared from the Rh2(acac)2 counterpart and Me3SiI and shown crystallographically to contain the PMe3 ligand in a truly symmetrical bridging position; a new synthetic route to migrate more bulky phosphines such as PPh3 and PiPr3 from a terminal into a bridging site is also described. PMID- 12778705 TI - A facile synthesis of bimodal mesoporous silica and its replication for bimodal mesoporous carbon. AB - Bimodal mesoporous silica material composed of 30-40 nm sized nanoparticles with 3.5 nm sized three-dimensionally interconnected mesopores was synthesized under neutral conditions using sodium silicate as a silica source. Using the bimodal mesoporous silica as a template, bimodal mesoporous carbon having 4 nm sized framework mesopores and approximately 30 nm sized textural pores was synthesized. PMID- 12778706 TI - Synthesis and crystallographic structure of a novel photoresponsive azobenzene containing organosilane. AB - A novel photoresponsive azobenzene-containing organosilane was synthesized via an isocyanato-amino coupling reaction, and its crystal structure was determined by X ray crystallography. PMID- 12778707 TI - Synthesis and characterization of highly ordered functional mesoporous silica thin films with positively chargeable -NH2 groups. AB - Highly ordered mesoporous organic-inorganic hybrid silica thin films with covalently bonded, positively chargeable -NH2 terminal groups were synthesized by evaporation induced self-assembly of tetraethoxysilane, 3-aminopropyl triethoxysilane, and a nonionic surfactant under acid conditions and characterized using TEM, GISAXS, FTIR, SAW-based N2 sorption, and TGA. PMID- 12778708 TI - First helical zinc(II) complex with a salen ligand. AB - The structure of a tetra-coordinated zinc(II) complex with a salen ligand was determined for the first time; unexpectedly, the complex was an interesting 2:2 metal-to-ligand complex. PMID- 12778709 TI - Stereoselective aza-Diels-Alder reactions with 2H-azirines as dienophiles furnishing highly functionalized tetrahydropyridines. AB - Highly diastereoselective Lewis acid mediated aza-Diels-Alder reactions of chiral auxiliary derivatized 2H-azirines have been accomplished for the first time, yielding bi and tri-cyclic heterocyclic compounds, comprising aziridine and tetrahydropyridine substructures, in up to 97% de; with the absolute stereochemistry of the major product confirmed by X-ray crystallography. PMID- 12778710 TI - A ruthenium dipyridophenazine complex that binds preferentially to GC sequences. AB - Uniquely, a Ru11 complex of the dppz ligand shows a preference for GC sequences of DNA. PMID- 12778711 TI - Chirality control in 2,2'-biphosphole ligand leading to enantiopure Pd complex. AB - Asymmetric alkylation of the 3,4-dimethyl-5-phenyl-2,2'-biphospholyl anion with the (2R,4R)-(-)-pentaneditosylate leads to a new chirally flexible 2,2' biphosphole ligand as a mixture of three diasteroisomers. By complexation with Pd(II), a chirality control occurs to afford enantiopure Pd complex. PMID- 12778712 TI - Isolation of electronic from geometric contributions to Bergman cyclization of metalloenediynes. AB - Conformationally constrained ethylene-diamine metalloenediyne compounds exhibit alkyne termini separations that are constant and independent of metal center geometry. Ancillary chloride ligand electron donation into the Bergman cyclization reaction coordinate, however, dramatically influences the observed temperatures. PMID- 12778713 TI - First report of Zn2+ sensing exclusively at mesoscopic interfaces. AB - We present a simple but unique, new probe 1-pyrenyl-methyl-bis(2-picolyl) amine (Pybpa) that selectively senses interfacially bound Zn2+ ions while being completely silent towards Zn2+ in bulk water. PMID- 12778714 TI - Biarylpyrimidines: a new class of ligand for high-order DNA recognition. AB - Biarylpyrimidines bearing omega-aminoalkyl substituents have been designed as ligands for high-order DNA structures: spectrophotometric, thermal and competition equilibrium dialysis assays showed that changing the functional group for substituent attachment from thioether to amide switches the structural binding preference from triplex to tetraplex DNA; the novel ligands are non-toxic and moderate inhibitors of human telomerase. PMID- 12778715 TI - Photoaccelerated oxidation of chlorinated phenols. AB - Exposure to visible light increases the rate of oxidation of chlorinated phenols by hydrogen peroxide in aqueous solution in either the presence or the absence of iron-based catalysts, which may be explained by the aqueous photoreactions of chloroquinone intermediates. PMID- 12778716 TI - A stable alkyl hydride of a first row transition metal. AB - Hydrogenolysis of a Cr(III) dialkyl precursor produced a binuclear chromium complex with a bridging hydride and a bridging alkyl; this structurally characterized organometallic compound is thermally very stable and does not undergo the expected reductive elimination of alkane. PMID- 12778717 TI - Studies of the functionalisation of organic-inorganic hybrid materials by using the Heck reaction. AB - Cogelation of 4-styryltrimethoxysilane was performed using NH4F as catalyst. The functionalisation of the styryl group was then studied by using Heck's reaction with ethyl 4-bromobenzyl- or 4-bromoarylvinylbenzyl-phosphonates. The efficiency of the solid-phase reaction was highly dependent on the texture of the solids. PMID- 12778718 TI - [Ni(eta 3-CH2C(CH3)CH2)(SbPh3)3][BAr'4]: an extremely active cationic allyl nickel-stibine catalyst for the oligomerization of styrene. AB - The pseudotetrahedral, formally 5-coordinate complex [Ni(eta 3 CH2C(CH3)CH2)(SbPh3)3][BAr'4] (Ar' = 3,5-C6H3(CF3)2) as well as the 4-coordinate derivative [Ni(eta 3-CH2C(CH3)CH2)(AsPh3)2][BAr'4] act as extremely efficient catalysts for the oligomerization of styrene. PMID- 12778719 TI - First total synthesis of the neuronal cell protecting carbazole alkaloid carbazomadurin A by sequential transition metal-catalyzed reactions. AB - The highly oxygenated neuronal cell protecting carbazole alkaloid carbazomadurin A was synthesized in nine steps and 11% overall yield from isovanillic acid. PMID- 12778720 TI - Copper and manganese: two concordant partners in the catalytic oxidation of p cresol to p-hydroxybenzaldehyde. AB - Copper and manganese were found to be two concordant partners in the synthesis of p-hydroxybenzaldehyde from p-cresol; under mild conditions, this research realised 95.6% selectivity for p-hydroxybenzaldehyde at 98.5% conversion of p cresol. PMID- 12778721 TI - New mode of sterically imposed phosphorus hyperco-ordination. AB - The sterically imposed electronic interaction in Nap (POCl2)(PCl4) (Nap = naphthalene-1,8-diyl) results in hyperco-ordination of the P atom by the O donor in the bridging position between the two peri-substituents. PMID- 12778722 TI - Cationic benzyl zirconium heteroscorpionates: synthesis and characterization of a novel ethylene polymerisation catalyst showing an unusual temperature dependent polymerisation mechanism. AB - The reaction of (bpzmp)Zr(CH2Ph)3 with B(C6F5)3 produces the active ethylene polymerisation catalyst [(bpzmp)Zr(CH2Ph)2]+[PhCH2B(C6F5)3]- which showed a temperature dependent polymerisation mechanism identified by variable temperature 1H NMR analysis of the catalyst solution. PMID- 12778723 TI - A new strategy towards ultra stable mesoporous titania with nanosized anatase walls. AB - A new and generally applicable synthesis procedure is developed in order to synthesise micelle-templated mesoporous titania built up of nanosized anatase walls with thermal stability up to 600 degrees C. PMID- 12778724 TI - Well-defined star-shaped calcite crystals formed in agarose gels. AB - Single crystals of calcite exhibiting a morphology of well-defined 8-armed stars, which evolved from original rhombohedral calcite crystals with their 8 points extending radially into eight arms, were produced by crystallization of CaCO3 in agarose gels. PMID- 12778725 TI - Homolytic 1,5-transfer of chiral organosilicon groups from an enoxy oxygen to an alkoxy oxygen--implications for mechanism. AB - Reaction of the optically active silanes, ((Ssi)-(-)-6), formed by treatment of racemic 2-methylenecycloheptanone oxide with LDA followed by (R)-(+) chloromethyl(1-naphthyl)-phenylsilane, with tributyltin hydride under standard radical conditions affords (2R/2S)-[(S)-(methyl(1-naphthyl) phensylsilyloxy)methyl]cycloheptanone, (Ssi)-(-)-7, providing strong evidence that homolytic 1,5-transfers of organosilicon groups from enoxy oxygen to alkoxy oxygen proceed with retention of configuration, most likely through a frontside attack mechanism rather than via a hypervalent intermediate. PMID- 12778726 TI - Halogen-induced selectivity in heterogeneous epoxidation is an electronic effect- fluorine, chlorine, bromine and iodine in the Ag-catalysed selective oxidation of ethene. AB - Selectivity promotion in the Ag-catalysed heterogeneous epoxidation of ethene correlates with halogen electron affinity showing that it is an electronic phenomenon rather than a steric or geometrical effect. PMID- 12778727 TI - Towards "designer" surfaces: functionalisation of self-assembled monolayer (SAM) on colloidal gold by alkene metathesis. AB - A variety of groups like a Fischer carbene complex, an N-hydroxysuccinimide or a ferrocene derivative have been grafted by ruthenium-catalyzed cross-metathesis reaction with terminal alkene groups on monolayer-protected gold clusters as a mild and convenient strategy to anchor functional molecules. PMID- 12778728 TI - A novel bioassay for screening and quantification of taxanes. AB - We report a novel method for screening and quantifying tubulin-binding antimitotic agents that promote microtubule formation. The method is based on the shift in the peak of the fluorescence emission difference spectrum of tubulin complexed to metal free tetrakis(4-carboxyphenyl)porphyrin (TCPP) in the presence of antimitotic agents. Taxol (paclitaxel), an anti-tumor drug approved for the treatment of a variety of cancers, caused the appearance of a new fluorescence peak at 645 nm at concentrations as low as 125 nM, the intensity of which was a function of the paclitaxel concentration. Cephalomannine caused the induction of a new fluorescence peak at 651 nm only above 1 muM. Baccatin did not induce the appearance of any new peak within detectable operating measurement conditions. These observations are in accordance with the biological activities/cytotoxicities of these compounds. Accordingly, it is proposed that the new method can be used for high throughput screening of antimitotic compounds. PMID- 12778729 TI - Two types of intramolecular homolytic substitution reactions at group XIV atoms: unusual radical 1,4-Sn shifts from Si to C and carbonylative SHi reaction at Si. AB - 4-[(Trimethylstannyl)diphenylsilyl]butanoyl radical, arising from the corresponding 3-(stannylsilyl)propyl radical and CO, undergoes an SHi reaction at Si with extrusion of trimethyltin radical to give silacyclopentanone. The parent 3-(stannylsilyl)propyl radical was also found to isomerize to (3 stannylpropyl)silyl radical via a 1,4-Sn shift from Si to C with a rate constant of 9.3 x 10(4) s-1 at 80 degrees C. Ab initio and DFT MO calculations support a front-side attack mechanism. PMID- 12778730 TI - Reductive cleavage of the C-O bond of acetals and orthoesters: reduction by silane in the presence of a Rh-PPh3 complex. AB - Reductions of acetals to ethers and of orthoester to acetal by hydrosilane using rhodium catalyst are described. PMID- 12778731 TI - Synthesis, spectroscopic characterisation and structure of the first chiral heteroleptic bis(phthalocyaninato) rare earth complexes. AB - Treatment of MIII(Pc)(acac) (M = Sm, Eu, Gd; Pc = phthalocyaninate; acac = acetylacetonate), generated in situ, with 3-(3-pentyloxy)phthalonitrile in the presence of 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) in n-pentanol affords racemic mixtures of the chiral double-deckers MIII(Pc)[Pc(OC5H11)4] [Pc(OC5H11)4 = 1,8,15,22-tetrakis(3-pentyloxy)phthalocyaninate], which have been spectroscopically and structurally characterised. PMID- 12778732 TI - Control of Al distribution in ZSM-5 by conditions of zeolite synthesis. AB - Concentration of Al-O-(Si-O)1,2-Al sequences located in one ring and forming cationic sites for divalent cations in ZSM-5 can be controlled in a wide range of their concentrations by the variation of the source of aluminium and silicon used for synthesis of this zeolite. PMID- 12778734 TI - Photochemical isomerization of 1,2,5-trisilabicyclo[3.2.0]hepta-3,6-diene to 1,4,7-trisilabicyclo[2.2.1]hepta-2,5-diene. AB - Upon irradiation of a benzene-d6 solution of 1,2,2,5-tetrakis[di-tert butyl(methyl)silyl]-4,7-diaryl- 1,2,5-trisilabicyclo-[3.2.0]hepta-3,6-diene [1a: aryl = phenyl, b: aryl = 3,5-bis-(trimethylsilyl)phenyl], 1,4,7,7-tetrakis[di tert-butyl-(methyl)silyl]-2,5-diaryl-1,4,7- trisilabicyclo[2.2.1]hepta-2,5-diene (2a,b) was formed via skeletal rearrangement. PMID- 12778733 TI - Biocatalytic reduction of ketones by a semi-continuous flow process using supercritical carbon dioxide. AB - The immobilized resting-cell of Geotrichum candidum was used as a catalyst for the reduction of a ketone in a semi-continuous flow process using supercritical carbon dioxide for the first time; it was also applied for the asymmetric reduction of a ketone and resulted in excellent enantioselectivity (ee > 99%) and a higher space-time yield than that of the corresponding batch process. PMID- 12778735 TI - A novel dry route to ortho-functionalized triarylbismuthanes that are difficult to access by conventional wet routes. AB - When an aryl iodide bearing an electron-withdrawing group at the ortho position was milled together with bismuth shots and calcite grains in the presence of Cu powder and CuI using a laboratory ball mill, the corresponding ortho functionalized triarylbismuthane was obtained in moderate to good yield. PMID- 12778736 TI - Ion-exchange synthesis and magneto-optical spectra of colored magnetic thin films composed of metal(II) hexacyanochromate(III). AB - Magnetic thin films composed of hexacyanochromate-based magnets, MII1.5[CrIII(CN)6].ZH2O (M = Co, Ni, Cu), were prepared on a transparent Nafion membrane by an ion-exchange process and their Faraday spectra were observed in the visible region. PMID- 12778737 TI - Preparation of novel polythioether dendrons on a solid support. AB - A synthetic scheme for the solid-phase synthesis of unprecedented polythioether dendrons has been established, the dendrons prepared up to the fourth generation, and the applicability of the dendronized resins for supported catalysis has been demonstrated. PMID- 12778738 TI - [Resveratrol promote permeability transition pore opening mediated by Ca2+]. AB - AIM: To investigate the mechanisms of anti-cancer effect of resveratrol (Res), and the effects of Res in cell apoptosis. The role of Res playing in mitochondrial permeability transition pore (PTP) induction was studied. METHODS: Mitochondria was prepared from the liver of Wistar rats. The effects of Res on oxygen consumption of isolated mitochondria from rat liver was measured with Clark-type electrode and resulted in respiration control rate (RCR). Mitochondrial swelling affected by Res was assessed spectrophotometrically, through the changes in absorbance at 540 nm. The PTP opening was learned from the results. Membrane potential of mitochondia was measured through fluorescence spectrophotometry. RESULTS: Res was shown to inhibit the respiration and decrease the RCR of mitochondria. Res can promote the PTP opening mediated by Ca2+. Res was shown to promote the increase of mitochondial membrane potential mediated by Ca2+ and loss of mitochondial membrane potential. CONCLUSION: Res was shown to inhibit mitochondial respiration and induce PTP opening of mitochondria. These may be one of the pathways that Res showed anti-cancer action and induce cells apoptosis. PMID- 12778739 TI - [Antioxidant effect of Ganoderma polysaccharide peptide]. AB - AIM: To study the antioxidant effect of Ganoderma polysaccharide peptide (GLPP) and its mechanism. METHODS: Copper was used as oxidant to induce low lipoprotein (LDL) oxidative modification, and alloxan was given i.v. to induce reactive oxygen species (ROS) injury in mice. RESULTS: GLPP decreased oxidation of LDL and the relative electrophoretic mobility (REM) of oxidative product of LDL. After GLPP was given i.p. for 20 days, the concentration of malondialdehyde(MDA) in serum and heart of mice was decreased. The GSHpx enzyme activity was increased, while the SOD level was decreased. The catalase(CAT) levels were not significantly changed by GLPP. CONCLUSION: GLPP showed antioxidant effect by scavenging ROS or enhancing the enzyme activity of GSHpx in vivo and in vitro. PMID- 12778740 TI - [A new vomiting animal model--mink]. AB - AIM: To establish a new, reliable vomiting model in minks. METHODS: Adult male minks (Mustela vison) were randomly divided into groups (n = 6). Cisplatin, apomorphine, copper sulfate and X-radiation were used to establish vomiting model. Retching and vomiting were observed after the vomiting models were given anti-vomiting agents. After the behavioral experiment, assay of 5-HT in the ileum was performed by immunohistologic method. RESULTS: Cisplatin 7.5 mg.kg-1 i.p., apomorphine 1.6 mg.kg-1 s.c. and copper sulfate 40 mg.kg-1 ig were shown to evoke vomiting. Retching and vomiting were significantly inhibited in ondansetron and metoclopramide pretreated minks (P < 0.05, P < 0.01). CONCLUSION: As a new vomiting model, minks may be of great value in studying vomiting mechanism and screening new antiemetic drugs. PMID- 12778741 TI - Comparison of the effects of polysaccharides from wood-cultured and bag-cultured Ganoderma lucidum on murine spleen lymphocyte proliferation in vitro. AB - AIM: To compare the influences of wood-cultured Ganoderma lucidum polysaccharides (Gl-PS-WC) and bag-cultured Ganoderma lucidum polysaccharides (Gl-PS-BC) on the proliferation activities of murine spleen lymphocytes in vitro, and investigate whether Gl-PS-BC can be substituted for Gl-PS-WC. METHODS: Mixed lymphocyte culture (MLC) reaction, lymphocyte proliferation induced by concanavalin A (Con A, 1 mg.L-1) or lipopolysaccharide (LPS, 5 mg.L-1), MLC reactions inhibited by immunosuppressive drugs, cyclosporine A (CsA, 0.1 mg.L-1), mitomycin (Mit C, 0.1 mg.L-1), or antitumor drug, etoposide (VP-16, 0.1 mg.L-1), were detected in the presence or absence of Gl-PS-WC and Gl-PS-BC in the concentration range of 0.2 12.8 mg.L-1. RESULTS: Two kinds of polysaccharides were shown to promote MLC in the range of 0.2-12.8 mg.L-1, increase lymphocyte proliferation induced by Con A or LPS and antagonize the inhibitory effects of CsA, Mit C or VP-16 on MLC. No significant difference was observed between these two kinds of polysaccharides in selected concentrations. CONCLUSION: Gl-PS-WC and Gl-PS-BC showed similar effects on the proliferation activities of murine spleen lymphocytes in vitro. PMID- 12778742 TI - [Effect of ginkgolide B on platelet-activating factor induced activation of rat polymorphonuclear leukocytes]. AB - AIM: To investigate the effect of ginkgolide B on PAF-induced adhesion, chemotaxis and degranulation of rat polymorphonuclear leukocytes (PMNs). METHODS: The adhesion of rat PMNs to rat synovial cells (RSC) was measured with MTT colorimetry. The chemotaxis of PMNs was quantified with Boyden chamber method. The degranulation of rat PMNs was evaluated by determining the activity of released beta-glucuronidase. RESULTS: In comparison with control, ginkgolide B at the concentration of 10 mumol.L-1 significantly inhibited the adhesion of PMNs to RSC by 71.74%. At the final concentration of 1-1,000 nmol.L-1, ginkgolide B dose dependently inhibited the chemotaxis of PMNs stimulated with 10 nmol.L-1 platelet activating factor (PAF), the IC50 was 4.84 nmol.L-1. At the final concentration of 0.01-10 mumol.L-1, ginkgolide B decreased the release of beta-glucuronidase in PMNs induced by 1 mumol.L-1 PAF in dose-dependent manner. The IC50 was 3.56 mumol.L-1. CONCLUSION: Ginkgolide B was found to significantly inhibit PAF induced adhesion, chemotaxis and degranulation in rat polymorphonuclear leukocytes. These effects might be considered a part of the mechanisms underlying the antiinflammatory action of ginkgolide B. PMID- 12778743 TI - [Protective effect of puerarin on vascular endothelial cell apoptosis induced by chemical hypoxia in vitro]. AB - AIM: To study the effect of puerarin on vascular endothelial cells apoptosis induced by chemical hypoxia-ischemia in vitro. METHODS: The chemical hypoxia ischemia model was performed by treating cultured bovine aortic endothelial cells (BAECs) with NaCN in glucose-free medium. Cell viability was determined by trypan blue staining. Cell apoptosis was defined by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL), flow cytometry and Hoechst 33342 staining. The expression of Caspase-3 in endothelial cells was detected by immunocytochemical method. RESULTS: Chemical hypoxia-ischemia was shown to initiate bovine aortic endothelial cell apoptosis. Puerarin (0.5-3 mmol.L-1) was found to inhibit endothelial cell apoptosis effectively, and reduce the expression of Caspase-3 significantly. CONCLUSION: Puerarin can protect apoptotic endothelial cells induced by chemical hypoxia-ischemia markedly and the effect was performed partly by decreasing Caspase-3 expression. PMID- 12778744 TI - [Prognosticating the region-selectivity in the O-methylation reaction of erythromycins by conformational search model]. AB - AIM: In order to find a way to prognosticate the region-selectivity in the O methylation reaction of erythromycin and its derivatives, conformation search model of Hyperchem Pro 6.0 used in personal computer was used to establish it. METHODS: The results of O-methylation reaction of compound 1, 2 and 3 showed the difference between 6-OH and 11-OH. Using the conformational search model of hyperchem, 1,000 conformations of compound 4, 5 and 6 were found with the parameter of dihedral angle. The 14-membered lactone ring was defined as a "plane". The position of carbonyl of 1- or 9- and hydroxyl of 6- or 11- is different, either "up-plane" or "down-plane". A statistic analysis of low-energy conformation clusters was used to sort these clusters by the parameter of the dihedral angle. The data is in accord with the results of the experiments. RESULTS: Conformation numbers, energy minimum, energy average and position of the active groups of compounds 4, 5 and 6, the simplified structures of compound 1, 2 and 3, were given in Table 1. by sorting the dihedral angle. CONCLUSION: The interaction of the hydroxyl and carbonyl were analysed and found that O methylation reactions of erythromycin and its derivatives were impacted by the space factor. The clusters of energy minimum, maximum number and "outside" were discussed. The selectivity of the O-methylation reaction of erythromycin and its derivatives can be prognosticated by analysing the parameter of the dihedral angle. PMID- 12778745 TI - [A new chromone glycoside from Cassia siamea lam]. AB - AIM: To study the chemical constituents of the stem of Cassia siamea. METHODS: The compounds were isolated by chromatography on silica gel, and identified on the basis of spectral analysis. RESULTS: Five compounds were isolated and identified as: beta-sitosterol (I), sucrose (II), n-octacosanol (III), 2-methyl-5 (2'-hydroxypropyl)-7-hydroxy-chromone-2'-O-beta-D-glucopyranoside (IV) and piceatannol (V). CONCLUSION: Compound IV is a new compound. Compounds II, III and V were obtained from this plant for the first time. PMID- 12778746 TI - [Isolation and structural identification of phenylethanoid glycosides from Corallodiscus flabellata]. AB - AIM: To study the chemical constituents from Corallodiscus flabellata. METHODS: The compounds were isolated and purified by macroporous adsorption resin, silica gel column chromatography and identified on the basis of their physiochemical and spectral data. RESULTS: Three phenylethanoid glycosides (I-III) were obtained from the n-BuOH fraction of water-extracts. Their structures were elucidated as 3,4-dihydroxyphenylethanol-8-O-[beta-D-apiofuranosyl (1-->3)]-beta-D glucopyranoside (I), 3,4-dihydroxyphenylethanol-8-O-[4-O-trans-caffeoyl-beta-D apiofuranosyl (1-->3)-beta-D-glucopyranosyl-(1-->6)]-beta-D-glucopyranoside (II) and 3,4-dihydroxyphenylethanol-8-O-[beta-D-apiofuranosyl(1-->3)-beta- D glucopyranosyl-(1-->6)]-beta-D-glucopyranoside (III). CONCLUSION: Compounds I, II and III are new compounds. PMID- 12778747 TI - [Determination of adefovir in monkey plasma by liquid chromatography-tandem mass spectrometry]. AB - AIM: To develop a sensitive and specific liquid chromatography-tandem mass spectrometry (LC/MS/MS) method for the determination of adefovir in monkey plasma. METHODS: Adefovir and internal standard 9-(3-phosphonylmethoxypropyl) adenine were isolated from plasma by protein precipitation with methanol, then chromatographed by using a Diamonsil C18 column. The mobile phase consisted of methanol-water-formic acid (20:80:1). Electrospray ionization (ESI) source was applied and operated in the positive ion mode. Selected reaction monitoring (SRM) mode with the transitions of m/z 274-->m/z 162 and m/z 288-->m/z 176 were used to quantify adefovir and the internal standard, respectively. RESULTS: The linear calibration curve was obtained in the concentration range of 0.02-4.00 mg.L-1. The lower limit of quantitation was 20 micrograms.L-1. The inter- and intra-day precision (RSD) was less than 5.8%, and the accuracy (relative error) was within +/- 4.5%. The method was successfully used in a pharmacokinetic study of adefovir dipivoxil in monkeys. CONCLUSION: The method is proved to be suitable for pre clinical investigation of adefovir dipivoxil pharmacokinetics, which offers advantages of specificity and simple sample preparation compared with the previously reported methods. PMID- 12778749 TI - [Study on separation of sulfonamides by capillary high-performance liquid chromatography and electrochromatography]. AB - AIM: To establish separation methods of five sulfonamides by using capillary high performance liquid chromatography(mu-HPLC) and electrochromatography. The effect of mobile phase varies such as methanol content, pH, buffer solution concentration and voltage on their chromatographic behavior and electroosmesis flow was investigated. Capillary electrochromatography (CEC) was compared with mu HPLC at the same condition. METHODS: Stationary phase was ODS, mobile phase was methanol and 2 mmol.L-1 H3PO4 buffer solution (pH 3.0-7.0), voltage was 0- -15 kV, flow rate was 10 microL.min-1, pressure was approximately 70 MPa and UV detection wavelength was 254 nm. RESULTS: Separations on base line have been respectively accomplished for five sulfonamides by mu-HPLC with mobile phase of methanol-2 mmol.L-1 H3PO4 buffer solution (30:70) at pH 5.0 in 67 min, and CEC with the same mobile phase at -5 kV voltage in 25 min. CONCLUSION: Electroosmesis flow of CEC decreased with the increase in methanol content, buffer solution concentration, increased with the increase in voltage and increase slightly with the increase in pH of mobile phase. Retention values (k) of solutes to be examined decreased with increasing methanol content of mobile phase in mu-HPLC and CEC. Retention values (k) of solutes increased slightly with increasing buffer solution concentration, decreased with increasing voltage in CEC. Trimethoprim(TMP) decreased obviously with increasing voltage in CEC. The effect of pH of mobile phase on retention values (k) was more complex. Five sulfonamides were separated at the same mobile phase condition by mu-HPLC and CEC. And separation speed of CEC was much faster than that of mu-HPLC. CEC was very fit for rapid separation of sulfonamides. PMID- 12778748 TI - [Determination of 9-nitrocamptothecin in rat plasma and tissues by high performance liquid chromatography]. AB - AIM: To develop a sensitive, specific and accurate method for quantifying 9 nitrocamptothecin in rat plasma and tissues and to study the distribution of 9 nitrocamptothecin in rat tissues. METHODS: Plasma and tissue samples were prepared based on a simple liquid-liquid extraction and separation through a Hypersil BDS C18 column. The mobile phase for plasma samples and tissue samples consisted of a mixture of acetonitrile-water-formic acid (35:65:2) and a mixture of acetonitrile-water-formic acid (30:70:2), respectively. The UV detector was set at 370 nm. RESULTS: A linear calibration curve of 9-nitrocamptothecin in plasma was obtained in the concentration range of 25-1,600 micrograms.L-1, and the quantitation limit of plasma and tissues was 25 micrograms.L-1. A linear range of concentrations for 9-nitrocamptothecin in heart, lung, spleen, stomach, fat, womb, and ovary was 10-1,000 ng.g-1, and the quantitation limit was 10 ng.g 1. A linear range of concentrations for 9-nitrocamptothecin in brain, kidney, liver, intestine, smooth muscle, skeletal muscle, and tectical was 5-500 ng.g-1, and the quantitation limit was 5 ng.g-1. The intra- and inter-run precision was measured to be below 11%. The inter-run accuracy was less than 5% for the analyte. After i.v. administration of 9-nitrocamptothecin, the drug was distributed extensively in rat in vivo. The concentration in lung was the highest, and the drug was accumulated in lung and liver. Following ig administration, the concentration in stomach was higher than that in other organs. CONCLUSION: The method is shown to be accurate and convenient, and suitable for preclinical pharmacokinetic studies of 9-nitrocamptothecin. PMID- 12778750 TI - [Preparation of cisplatin multivesicular liposomes and release of cisplatin from the liposomes in vitro]. AB - AIM: To prepare cisplatin multivesicular liposomes with high encapsulation efficiency and sustained-release character, and compare the release characteristics with conventional liposomes prepared by reverse-phase evaporation method. METHODS: Cisplatin multivesicular liposomes were prepared using multiple emulsion method. The concentrations of cisplatin and lipids in the liposomes were measured by flameless atomic absorbance spectroscopy (FAAS) and phosphalipid enzyme reagent method, respectively. The encapsulation efficiency, size and release of the cisplatin from the liposomes were studied in vitro. RESULTS: The mean diameter of cisplatin multivesicular liposomes was (16.6 +/- 1.0) micron. The encapsulation efficiency of cisplatin was more than 80%. The release profile in vitro fitted with a first-order equation. The releasing t1/2 of cisplatin multivesicular liposomes is 37.7 h, which is 8.4 that of conventional liposomes. Co-membrane stabilizer has remarkable stabilizing effect on the multivesicular liposomal membrane confirmed by differential scattering calorimetry (DSC). CONCLUSION: The cisplatin multivesicular liposomes showed high encapsulation efficiency and sustained-release character. PMID- 12778751 TI - [Hypoglycemic effect of polysaccharide-coated insulin liposomes after oral administration in mice]. AB - AIM: To evaluate the hypoglycemic effect of chitosan-coated and sodium alginate coated insulin liposomes after oral administration in mice. METHODS: Insulin liposomes were prepared by reverse-phase evaporation. Chitosan and alginate coating was carried out by mixing liposomal suspension with chitosan and sodium alginate solutions, followed by incubation. The particle size and morphology of insulin-liposomes were determined using laser light scattering instrument and transmission electron microscopy (TEM). The entrapment efficiency was analyzed using HPLC and ultracentrifuge. The protection of insulin from peptic and tryptic digestion was studied with HPLC. The hypoglycemic effects of polysaccharide coated insulin liposomes were investigated using the glucose oxidase method after oral administration in mice. RESULTS: The particle size of uncoated, chitosan coated and alginate-coated insulin-liposomes was (138 +/- 31) nm, (230 +/- 20) nm and (266 +/- 19) nm, respectively. All insulin-liposomes were of spherical or ellipsoidal shape. The entrapment efficiencies were 81.6%, 73.5% and 68.7%, respectively. Insulin was protected from tryptic digestion by chitosan-coated liposomes and protected from peptic digestion by alginate-coated liposomes. The hypoglycemic effects of insulin-liposomes, coated with 0.1% chitosan and 0.1% sodium alginate, were observed. CONCLUSION: Chitosan-coated and sodium alginate coated liposomes were shown to reduce peptic or tryptic digestion on insulin, and enhance enteral absorption of insulin. PMID- 12778752 TI - [Study on the hepatocytic cell targetability of liposomes]. AB - AIM: To target for hepatocytic cell, liposomes was modified by special ligand. METHODS: Sterically stabilized liposomes (SSL) was conjugated with asialofeticin (AF), the ligand of asialoglycoprotein receptor (ASGP-R) of hepatocyte. ASGP-R BLM is the ASGP-R reconstructed on bilayer lipid membrane (BLM). The recognition reaction between AF-SSL and ASGP-R-BLM can be monitored by the varieties of membrane electrical parameters. The targetability of AF-SSL mediated to hepatocyte was detected by radioisotopic labeled in vitro and in vivo. The therapeutic effect of antihepatocarcinoma was observed also. RESULTS: The lifetime of ASGP-R-BLM decreased with the added amount of AF-SSL. It was demonstrated that there was recognition reaction between AF-SSL and ASGP-R-BLM. The combination of AF-SSL with hepatocyte was significantly higher than that of SSL without AF-modified in vitro and in vivo. The survival time of rat for AF-SSL carriered ADM (adriamycin) group was much longer and the toxicities on heart, kidney and lung were lower than those SSL carried ADM group. CONCLUSION: It is possible to actively target the cell with specific receptor by ligand modified liposomes. The result prvide scientific basis of hepatocyte targeted liposomes. PMID- 12778753 TI - [RbcL sequence analysis of Belamcanda chinensis and related medicinal plants of Iris]. AB - AIM: To identify "Shegan" [Belamcanda chinensis (L.) DC.] and relative medicinal plants of Iris including Iris tectorum Maxim., I. dichotoma Pall., I. germanica L. and I. japonica Thunb. by ribulose 1,5-bisphosphate carboxylase Large Gene (rbcL) sequence analysis. METHODS: General DNA was isolated from the fresh leaves of Belamcanda chinensis and 4 Iris spp. by CTAB. A pair of primers was designed to amplify the rbcL gene and PCR Preps DNA kit was used to purify the PCR products. The rbcL sequences were determined by ABI (Applied Biosystems Inco.) Prism 310 Genetic Analyzer. RESULTS: A fragment of about 750 bp of rbcL gene from Belamcanda chinensis and 4 Iris spp. were amplified and sequenced. The rbcL sequences of Iris tectorum, I. dichotoma Pall. and I. japonica were reported for the first time. The rbcL sequences of 5 species of Iridaceae were aligned and analyzed using Clustal (Version 8.0) and MEGA (Version 2.0.) programs. The nucleotide number of difference is from 1.000 to 20.000. The tranversions is from 0.000 to 9.000 and the transitions is from 0.000 to 14.000. Phylogenetic tree based on rbcL partial sequence data indicated that the eleven samples of 5 species clustered separately. CONCLUSION: The sequence variation of rbcL can be used to identify Belamcanda chinensis and 4 species of relative medicinal plants of Iris. The molecular phylogenetic tree accords with the classical taxonomy. PMID- 12778754 TI - [Solid phase micro-extraction technique and its applications in medical analysis]. PMID- 12778755 TI - [A review of 545 consecutive free flap transfers for head and neck reconstruction in a new microsurgery unit]. AB - OBJECTIVE: To analyze the value of free flap transfers in head and neck reconstruction. METHODS: Five hundred and forty-five consecutive free flap transfers performed in 507 patients from May 1999 to September 2002 were reviewed. Data concerning the operation included date of surgery, defect description and site, stage and histology of tumor, flap type, recipient vessel and complications. RESULTS: The free fibula flap was most commonly used, followed by free radial forearm flap, rectus abdominis flap, jejunum flap, scapular flap, illiac crest flap, latissimus dorsi flap, and anterolateral thigh flap. The overall success rate of flap was 98.2% (535/545). The overall complication rate was 25.2% (128/507). The vessel thrombosis rate was 4.8% (26/545), and the flap salvage rate was 61.5%(16/26). CONCLUSION: Free flap transfer in head and neck region is safe and reliable. It is superior to the conventional pedicled flap technique. PMID- 12778756 TI - [Application of frontal-lateral laryngectomy in laryngeal cancer treatment]. AB - OBJECTIVE: Thirty-eight patients of laryngeal cancer were treated by extended frontal-lateral laryngectomy and the results were evaluated. METHODS: Thirty eight patients with laryngeal cancer were treated by frontal-lateral laryngectomy and the laryngeal defects were reconstructed by epiglottic flap, bi-pedicle and bi-muscle flap and epiglottic flap plus bi-pedicle and bi-muscle flap from February 1988 to February 2000. There were 29 males and 9 females, ranging in age from 32 to 72 years old. Radical neck dissection was performed in 12 patients. 36 cases belonged to glottic cancer and 2 was superglottic cancer (T2 16, T3 22). RESULTS: The aspiration was light and the majority of cases began to eat in 10-15 days after operation. The decannulation rate was 97.3% (37/38). Wound infection occurred in 3 cases, but pharyngocutaneous fistula was not found. The three and five years survival rates were 86.1% (31/36) and 83.9% (26/31) respectively. CONCLUSION: This procedure present an ideal effect and is worthy to be recommended, but only the indication is limited. PMID- 12778757 TI - [Laryngeal function reconstruction with sternohyvoid muscle after partial laryngectomy]. AB - OBJECTIVE: To study the laryngeal function reconstruction with sternohyvoid muscle after partial laryngectomy to treat T2 phase of glottis cancer. METHODS: 66 patients diagnosed to be at T2 phase of glottis cancer were treated by partial laryngectomy from 1992 to 1998. At the same time, they were rebuilt vocal cords with sternohyvoid muscle. RESULTS: All patients were decannulated from 2 to 3 weeks. They renewed own respiration, swallow and voice function. Overall three year survival rate was 97.0% and five-year survival rate was 93.8%. CONCLUSION: New operation is superior than traditional one in improving survival rate and life quality. PMID- 12778758 TI - [Pharyagoesophageal sphinoter myotomy for voice rehabilitation after total laryngectomy]. AB - OBJECTIVE: In those patients received total laryngectomy, Blom-Singer prosthesis speech appears to be the more effective form of rehabilitation. To reduce the effect of pharyngoesophageal sphincter on the Blom-Singer prosthesis voice restoration following total laryngectomy, the pharyagoesphageal sphincter myotomy was designed and carried out. METHODS: After total laryngectomy, the cricopharyngeus muscle stricture or spasm appears to inhibit the air flowing and prevent the speech. The pharyngoesophageal sphincter is now defined as the area including the cricopharyngeus with a portion of inferior constrictor, as well as some of the upper cervical esophagus. The pharyngoesophageal sphincter were incised intra- and post- operation of total laryngectomy, the dissection was approximately 5 cm in vertical length and 1 cm in width. RESULTS: Among the 33 patients who received pharyngoesophageal sphincter myotomy following total laryngectomy, voice restoration was achieved in 32 cases, the success rate was 97%. 25 patients had survived for 3 years, 18 patients survived for 5 years. CONCLUSION: Pharyagoesophageal sphincter myotomy can raise the success rate of rehabilitation of the voice by Blom-Singer prosthesis. PMID- 12778759 TI - [Treatment of severe subglottic laryngotracheal stenosis using hyoid graft with sternohyoid muscle flap]. AB - OBJECTIVE: To evaluate the possibility and reliability of the hyoid-sternohyoid graft transfer in the correction of server subglottic laryngotracheal stenosis, and delineate the operation skills and clinical results. METHODS: Seven patients with severe subglottic stenosis underwent laryngotracheal reconstruction using the hyoid grafts with sternohyoid muscle flaps (HG-SHMF). Five of these patients had traumatic subglottic stenosis, one with scar tissue of unknown etiology arising in the subglottic region, another with tracheal narrowing caused by inhalation of hydrochloric acid. RESULTS: All seven patients were successfully decannulated with moderate good voice. The average time from reconstruction to decannulation was 15.4 months. The stent was endoscopically removed with a range of 3 to 22 months; the mean time required for stenting was 9.6 months. Two patients who received additional salvage reconstruction procedures because of graft or stent displacement were extubated with improved voices and satisfactory airway. CONCLUSIONS: The HG-SHMF transfer was a single-stage reconstruction, relatively simple procedure that can restore an adequate airway and a good voice. Patients undergoing laryngotracheal reconstruction with HG-SHMF must have regular, long-term follow-up since graft displacement and recurrent granulation tissue or scar reformation can cause restenosis after an initially successful surgery. This procedure should be used in a large number of patients to further test its reliability. PMID- 12778760 TI - [Surgical management of benign pharyngo-esophageal stenosis]. AB - OBJECTIVE: To study the methods and outcome of surgical management for pharyngo esophageal stenosis. METHODS: Twenty-seven patients with pharyngo-esophageal stenosis from January 1983 to June 2001 were reviewed. Among 27 cases, there were 12 cases accompanied with laryngeal stenosis and one case accompanied with tracheal stenosis. In terms of etiological factor, all patients were cataloged into two groups, i.e. 25 cases with chemo-causis and 2 cases with trauma. The repair methods included colon interposition in 20 patients, free jejunum transplantation in 6 patients and pectoralis major muculocutaneous flap in 1 patient. In total 12 cases of laryngeal stenosis, sternohyoid myofascial flap was applied in 8 cases, and sternohyoid myofascial flap and epiglottis were applied in 4 cases. RESULTS: Swallow function recovered in 25 cases and failed in 2 cases. Laryngeal function recovered totally in 10 patients and partially in 2 patients with laryngeal stenosis. The patient with tracheal stenosis recovered and decannulated. CONCLUSION: According to the foci of pharyngo-esophageal stenosis, the colon interposition, the free jejunum transplantation and the pectoralis major myocutaneous flap can be applied respectively to restore normal physiological function. PMID- 12778761 TI - [Construction of bicistronic eukaryotic vector containing basic fibroblast growth factor and study of their functions in gene therapy for hearing impairment]. AB - OBJECTIVE: To construct basic fibroblast growth factor(bFGR) and enhance green fluorescence protein(EGFP) fusion gene eukaryotic expression vector internal ribosome entry site (pIRES)-bFGF-GFP and to evaluate the effect of transduction bFGF gene on noise induced hearing-loss in inner ear hair cells of guinea pigs. METHODS: Human bFGF cDNA was inserted into mammalian expressed plasmid pIRES EGFP. The recombinant expression plasmid pIRES-bFGF-EGFP was transfected into inner ear of guinea pigs, using lipofectin method. The transduced bFGF gene was mediated by SA lipidsome. IRES- bFGF-GFP was administered into the round window as rescue agent at the same time of noise exposure or as a protective agent 7 days before. RESULTS: SA liposome-mediated bFGF expressed at a high level in the cochlea of guinea pigs, and in the rescue group, a significant lower hearing thresholds was displayed. pIRES- bFGF-EGFP could protect hair cells. It demonstrated that pIRES- bFGF-GFP could protect the inner ear both structurally and functionally. bFGF/EGFP gene could be transcripted and translated into inner ear hair cells of guinea pig. bFGF/EGFP gene could express a specific protein. The recombinant bFGF/EGFP had significant protective effect as well as manifestation of autonomous fluorescence. CONCLUSION: bFGF/EGFP fusion protein not only expressed in hair cells of guinea pig but also showed significant bFGF activity and autonomous fluorescence. IRES induced exogenous gene could enter the hair cells. PMID- 12778762 TI - [Intra-arterial thrombolysis for the treatment of restricted cochlear microcirculation disorder in guinea pigs]. AB - OBJECTIVE: To ascertain a new treatment method for restricted cochlear microcirculation disorder. METHODS: Photochemical reaction was utilized to induce localized microcirculation damage to the second cochlear turn of every guinea pig. The animals were divided into 5 groups. Group 1 was blank control. Group 2 and Group 3 were injected urokinase (UK) through left subclavian artery or left external jugular vein 30 minutes after photochemical reaction. Group 4 and Group 5 were two control groups. Instead of urokinase, saline injection was applied accordingly. RESULTS: Both intra-arterial thrombolysis and intravenous thrombolysis were effective to improve the hearing levels and the blood supply to the inner ear. 50 minutes after urokinase injection, animals with intra-arterial thrombolysis showed a lower action potential threshold than that with intravenous thrombolysis (P = 0.025). And this phenomenon lasted 30 minutes. Cochlear blood flow of the animals with intra-arterial thrombolysis began to decrease and action potential began to increase 80-90 minutes after UK injection. CONCLUSION: Both intra-arterial thrombolysis and intravenous thrombolysis were effective to the restricted thrombosis in the stria vascularis, and intra-arterial thrombolysis showed a better improvement in the early stage of thrombolysis. PMID- 12778763 TI - [Protective effect of daizein on apoptosis cells of nasal mucosas in ovariectomized rats]. AB - OBJECTIVE: To study the protective effect of daizein on apoptosis cells of atrophic nasal mucosas in ovariectomized rats. METHODS: Sixty rats were divided into four groups as contrary, ovariectomized, ovariectomized + nylestriol (O + N) and ovariectomized + daizein (O + D), each with 15 rats. Earlier apotosis cells of nasal mucosas taken from nasal septum were measured with flow cytometry. RESULTS: Compared with contrary group, the number of apoptosis cells of mucosas increased after being ovariectomized, the number of apoptosis cells of mucosas in O + N and O + D groups didn't change. CONCLUSION: Estrogen replacement and daizein might have effects on protecting cells of mucosas from lacking of estrogen by decreasing apoptosis cells in ovariectomized rats. PMID- 12778764 TI - [Role of expression inducible nitric oxide synthase-mRNA of leukocyte and nasal mucosa in allergic rhinitis]. AB - OBJECTIVE: To investigate the relationship between expression of leukocyte inducible nitric oxide synthase (iNOS)-mRNA and allergic rhinitis (AR). METHOD: Thirty-five patients with AR and 30 healthy controls were included in this study. Expression of iNOS-mRNA in peripheral blood leukocyte was detected by in situ hybridization. NO in plasm was measured by nitrate reductase. Expression of iNOS mRNA in nasal mucosal was detected in 8 patients with AR and 6 healthy controls. RESULT: No expression of leukocyte iNOS-mRNA in healthy controls was found. In AR patients, the positive cells were significantly increased, the positive rate reached 40.82%. Expression of iNOS-mRNA was localized at the epithelium, gland and macrophage in healthy controls. Hyperplasia and expression of iNOS-mRNA increased at epithelium, gland and macrophage in the AR patients(t = 23.17, P < 0.001). The level of plasm NO in AR group was higher than that in healthy control group (t = 27.89, P < 0.001). CONCLUSION: There is a relationship between expression of leukocyte iNOS-mRNA and the level of plasm NO in AR patients. The study provides an easy method of in situ hybridization for detecting some signal in body. PMID- 12778765 TI - [Adenovirus p16(INK4A) gene therapy for laryngeal squamous cell carcinoma]. AB - OBJECTIVE: To assess the anti-tumor effects of p16INK4A gene transfer in laryngeal squamous cell carcinoma. METHODS: A Complete p16INK4A gene was inserted into a replication-defective recombinant adenovirus (Ad-p16) and the tumor cells were infected with Ad-p16. Confirmation of p16INK4A protein expression after Ad p16 infection was performed by Western Blotting. The therapeutic effects were evaluated both in in vitro and in vivo study. RESULTS: The replication-defective recombinant adenovirus can direct a high level of p16INK4A protein expression in laryngeal squamous cell carcinoma. Studies both in in vitro and in vivo experiments demonstrated that Ad-p16 treatment significantly inhibits the cell growth and the established tumors in nude mice. The mean value of tumor volumes among the Ad-p16, Ad-Lacz and phosphatic buffered saline(PBS) groups was (91.00 +/- 6.32) mm3, (137.00 +/- 9.62) mm3 and (144.00 +/- 13.87) mm3 respectively. Statistic analysis showed that there was a significant difference between the Ad p16 group and the control groups (P < 0.05). No significant difference was found between Ad-lacZ and PBS groups, which indicated that the antitumor effects was not influenced by the adenovirus. CONCLUSION: These results demonstrate a significant antitumor effect of Ad-p16 against human laryngeal squamous cell carcinoma. This data also further support the potential application of Ad-p16 to treat the human head and neck cancer. PMID- 12778766 TI - [Expression and significance of the urokinase-type plasminogen activator and its inhibitors in squamous cell carcinoma of human larynx]. AB - OBJECTIVE: To detect the expression of urokinase-type plasminogen activator (uPA) and its inhibitors(plasminogen activator inhibitors, PAI) type-1 and type-2 in squamous cell carcinoma of human larynx and reveal the correlation of the major clinicopathologicl parameters and prognosis. METHODS: uPA, PAI-1 and PAI-2 were detected from 104 cases squamous cell carcinoma of human larynx undergoing primary resection using immunohistochemistry(labeled-streptoavidin-biotin peroxidase, SAB) method. The results were classified positive and negative. Patients were followed-up prospectively for a median of 41 months(rang 24 to 84 months). Overall survival were analyzed according to Kaplan-Meier and log-rank statistics, the prognostic relevance of uPA, PAI-1 and PAI-2 and conventional prognostic factors were analyzed by Cox analyses. RESULTS: uPA, PAI-1 and PAI-2 positivity were present both in neoplastic cells and in fibroblast cells and macrophages. However, depending on the histological grading and invasive capacity of the tumor, a pronounced intra- and intertumoral heterogeneity in uPA staining was observed. The total positive rate of uPA, PAI-1 and PAI-2 was 66.3%, 70.2% and 50.0% respectively. No relationship between the expression of these proteins and clinicopathological parameters except for lymph node metastasis and recurrence, P value was 0.010, 0.027 and 0.038 respectively. Univariate survival analysis revealed a high significant inverse correlation of uPA positive expression survival time. Patients with PAI-2 positive expression had a significantly longer survival time than those with PAI-2 negative expression. In different clinicopathological parameters subgroups, uPA, PAI-1 and PAI-2 added significant survival information. Multivariate analysis revealed that four independent prognostic factors for overall survival time were uPA, PAI-2, lymph node metastasis and recurrences and clinical stage, P = 0.001, 0.002, 0.035 and 0.005 respectively. CONCLUSION: These findings suggest that uPA play important role in the metastasis of human laryngeal carcinoma and PAI-2 appears to be a true inhibitor contrary to PAI-1. PAI-1 might act as an essential modulator of the plasminogen activation system or a protector of carcinoma tissue against self degradation rather than as a simple inhibitor of system. uPA and PAI-2 could be new independent and strong biologically prognostic factors. PMID- 12778767 TI - [Electrically evoked compound action potential thresholds of pediatric cochlear implant recipients and its clinical application]. AB - OBJECTIVE: To investigate the possibility of using pediatric cochlear implant mapping protocol, to estimate the psychophysical levels based on the electrically evoked compound action potential (ECAP) threshold measured with the neural response telemetry (NRT) capabilities of cochlear corporation's CI24M device. METHODS: ECAP amplitude growth functions were regressed via NRT 3.0 software to determine ECAP threshold. 6 pediatric cochlear implant recipients' ECAP thresholds 1, 2 and 3 months after surgery were compared, meanwhile, intraoperative and postoperative ECAP thresholds in 7 pediatric recipients were compared. RESULTS: The ECAP amplitude growth functions were not linear when ECAP was close to threshold or saturation. There were no significant difference among the ECAP thresholds 1, 2 and 3 months after surgery. The average intraoperative ECAP threshold of 22 electrodes was 15 CL greater than postoperative threshold. It showed high relationship between them (R2 = 0.9154). CONCLUSION: ECAP threshold should be determined by regression of data from linear part of amplitude growth function. Only one NRT trial should be assessed when mapping protocol based on ECAP threshold to estimate pediatric map parameter. The intraoperative ECAP threshold can act as C-level at the initial mapping. Key words cochlear implant, electrically evoked compound action potential, mapping. PMID- 12778768 TI - [Treatment of endonasal-encephalomeningocele by transnasal endoscopic surgery]. AB - OBJECTIVE: To inquire into the feasibility of transnasal endoscopic surgery for endonasal encephalomeningocele, and to put forward the basic management guidelines. METHOD: Five cases of endonasal encephalomeningocele were managed by transnasal endoscopic surgery. Different surgical techniques were applied according to the size and shape of skull base defect. At the same time, one-stage repair of skull base defect was achieved by applying muscle, fascia, cartilage and bone. RESULTS: All 5 patients were successfully treated without complication and relapse during 1-5 year follow-up. CONCLUSION: Endonasal type encephalomeningocele can be managed by transnasal endoscopic surgery. If the skull base defect is less than 0.5 cm, it can be repaired by packing muscle and fascia. If the defect is larger than 0.5 cm, the cartilage or bone should be used to repair the skull base. PMID- 12778769 TI - [Treatment of traumatic pseudoaneurysm through completely embolizing siphon section of internal carotid artery]. AB - OBJECTIVE: Pseudoaneurysm in siphon section of internal carotid artery (ICA) caused by craniocerebral trauma was diagnosed and totally embolized by applying digital subtraction angiography (DSA) technique, with satisfactory results. The clinical application of this technique was discussed. METHODS: The patients suffered from craniocerebral trauma with serious epistaxis were managed in our hospital by DSA. The patients with pseudoaneurysm in ICA were chosen from them. The circular function of Willis circulus in these patients were checked. If it was good, the ICA was totally embolised. RESULTS: Among 32 patients suffered from craniocerebral trauma with serious epistaxis, 6 patients were definitely diagnosed by DSA to have pseudoaneurysm. The embolism was performed in 5 of them. Among these 5 patients, 4 cured, and 1 died. The patient who could not be embolized also died. CONCLUSION: The pseudoaneurysm definitely diagnosed by DSA can be treated by embolizing siphon section of internal carotid artery completely, after checking the compensatory function of side branches in Willis circulus. PMID- 12778770 TI - [Clinical analysis and long-term follow up study of asymptomatic nasopharyngeal carcinoma patients]. AB - OBJECTIVE: To observe the character of Epstein-Barr(EB) virus serology, fibroscopy appearance and prognosis of asymptomatic nasopharyngeal carcinoma(NPC). METHODS: Viral capsid antigen's IgA (VCA/IgA) of EB virus and early antigen's IgA(EA/IgA) of EB virus were detected by immunoenzymatic method. The clinical examination was carried out, including indirect mirror examination and fibroscopy of the nasopharynx and multiple biopsies. All patients of NPC were followed up to the end of 1999. RESULTS: 1. The geometric mean titer of VCA/IgA and EA/IgA are 1:100.79 and 1:10.76 respectively when asymptomatic NPC was diagnosed. There were no significant difference between VCA/IgA and EA/IgA antibody titres of asymptomatic patients and symptomatic cases (P > 0.05). The survival rates in these asymptomatic cases were higher than symptomatic patients (P < 0.05). 2. There was no correlation with the VCA/IgA or EA/IgA titer and the prognosis (P > 0.05) and the cervical lymph node metastasis (P > 0.05) when NPC was diagnosed. CONCLUSION: This is helpful to detect asymptomatic NPCs by EB serological screening periodically and nasopharyngeal fibroscopy and multiple biopsies. PMID- 12778771 TI - [How to avoid hoarseness for operation of schwannoma of the cervical vagus nerve]. AB - OBJECTIVE: To avoid hoarseness for operation of schwannoma of the cervical vagus nerve (SVN). METHODS: From Jan. 1997 to Sept. 2001, ten cases of SVN were operated. The procedures were as follows: Exposing the tumor and the vagus nerve, confirming its origin from the fascicle of recurrent laryngeal nerve (RLN) or the fascicle of non-RLN, making an small incision on the posterolateral surface of the tumor, usually posterior to the internal jugular vein, and then tearing and enlarging the small incision to enucleate the tumor. RESULTS: Two of the four cases of SVN originated from the fascicle of RLN happened temporary hoarseness and other two without hoarseness. All six cases of SVN originated from the fascicle of non-RLN, but one, happened no hoarseness. CONCLUSION: Above-mentioned technique is a good and reliable method to avoid hoarseness in the operation of SVN. PMID- 12778772 TI - [Fabry disease: new clinical research--current therapeutic perspectives]. PMID- 12778773 TI - Implausible research: how much is enough? PMID- 12778776 TI - Distant healing--an "update" of a systematic review. AB - AIM: To update our published systematic review of clinical trials of distant healing. DESIGN: Systematic review. DATA SOURCES: Medline, Embase, Cochrane Library and personal files. STUDY SELECTION: Any type of clinical study of any type of distant healing published between 2000 and December 2002. DATA EXTRACTION: For each included study, essential data were extracted and summarised in narrative form. RESULTS: 8 non-randomised and 9 randomised clinical trials were located. The majority of the rigorous trials do not to support the hypothesis that distant healing has specific therapeutic effects. The results of two studies furthermore suggest that distant healing can be associated with adverse effects. CONCLUSION: Since the publication of our previous systematic review in 2000, several rigorous new studies have emerged. Collectively they shift the weight of the evidence against the notion that distant healing is more than a placebo. PMID- 12778775 TI - Anderson-Fabry disease in Austria. AB - Fabry disease is an X-linked inherited inborn error of glycosphingolipid catabolism. The deficiency of alpha-galactosidase A leads to the deposition of glycosphingolipids primarily in lysosomes of blood vessel cells. In classically affected hemizygotes clinical manifestations include pain in the extremities, vessel ectasia (angiokeratoma) in skin and mucous membranes, ophthalmological abnormalities, and hypohidrosis. As disease progresses there is renal, cardiac, cerebral and vascular involvement, with most patients experiencing renal insufficiency, cardiac hypertrophy or stroke. Many female carriers of Fabry disease also have symptoms. Recently available enzyme replacement therapy has the potential to control or even reverse disease progression. The present analysis reports on five Austrian families with Fabry disease, cared for by nephrologists in June 2002. Furthermore we discuss potential indications for enzyme replacement therapy in patients maintained on renal replacement therapy. PMID- 12778774 TI - [Atherosclerosis and uremia: signifance of non-traditional risk factors]. AB - Arteriosclerosis, atherosclerosis and vascular calcification are causally related to the high morbidity and mortality of patients with chronic renal failure. Oxidative stress and carbonyl stress of uremia, dialysis procedure and/or intravenous iron therapy result in AGE (advanced glycation end-product), ALE (advanced lipoxidation end-product) and AOPP (advanced oxidation protein product) formation, favouring together with elevated CRP (C-reactive protein) levels the development of cardiovascular and cerebrovascular complications. Enhanced plasma levels of homocysteine and ADMA (asymmetric dimethylarginine) contribute to this process. In addition, in chronic renal insufficiency hyperphosphatemia and an enhanced calcium x phosphorus ion product are associated with the morbidity and mortality of the patients, particularly in the presence of fetuin deficiency. Phosphorus, AGEs and AOPPs, beside other factors, catalyze the conversion of vascular smooth muscle cells to osteoblast--like cells (particularly in the presence of monocytes/macrophages), resulting in bone matrix protein formation. Other risk factors, such as age, male sex, smoking, hypertension, diabetes, chronic inflammation, insulin resistance or dyslipidemia (enhanced non-HDL cholesterol) also contribute to the atherosclerotic risk profile of the patient with chronic renal insufficiency. While there is growing understanding of the mechanisms involved in arteriosclerosis, atherosclerosis and vascular calcification in uremia, we are still missing effective therapeutic maneuvers for reduction of excess mortality in uremic patients. PMID- 12778778 TI - Influence of tumor grade on expression of CA 125 in serum and tissue in patients with serous ovarian carcinoma. AB - OBJECTIVE: The factors affecting CA 125 expression in tissue in serous carcinoma are not well known. The aim of our study was to establish how CA 125 expression in serum and tissue is affected by DNA-ploidy, proliferative activity of tumor cells, tumor grade and the stage of the disease. STUDY DESIGN: This retrospective study included 50 patients with ovarian carcinoma. The serum CA 125 level was determined preoperatively in all patients. Antigen expression in tissue was confirmed using CA 125-immunohistochemistry. Suspensions of nuclei were prepared from tissue sections and DNA-ploidy was determined using flow cytometry. Data were analyzed using nonparametric statistics and multivariant analysis. RESULTS: The average level of serum CA 125 was 674.5 U/ml and was significantly higher in advanced (p = 0.00044) and dedifferentiated (p = 0.00055) tumors and in tumors with a higher S-phase fraction (SPF) (p = 0.011). The mean percent of CA 125 positive cells in tissue was 38.3. CA 125 immunostaining was not related to tumor grade or DNA-ploidy. A positive CA 125 test in both tissue and serum was confirmed in 33 cases (66%). The correlation between expression of CA 125 in tissue and serum was weak (r = 0.28) and nonsignificant (p = 0.052). CONCLUSIONS: No reliable conclusions could be drawn on the influence of the studied parameters on expression of CA 125 in tissue and serum. The effect of tumor grade seems to be the strongest influence, particularly on CA 125 expression in serum, but it is not independent. PMID- 12778777 TI - Quality of life in nonorganic and organic sleep disorders: I. Comparison with normative data. AB - Subjective health-related quality of life (HRQoL) was investigated in 100 patients with disturbed sleep (39 women aged 52 +/- 13 years and 61 men aged 53 +/- 10 years) referred to the sleep laboratory and compared with HRQoL in 100 normal healthy adults. Measurements included the Quality of Life Index (QLI) (Mezzich and Cohen), and objective (polysomnographic) and subjective (psychometric) quality of sleep and awakening. Statistical analysis (Mann-Whitney U-test) showed HRQoL to be significantly reduced in sleep disorders (SDs), with a more pronounced reduction in nonorganic than in organic SDs. Patients with nonorganic hypersomnia were more disturbed than those with nonorganic insomnia. Within organic SDs, patients with apnea were more disturbed than those with obstructive snoring. Out of ten elementary HRQoL components, seven were disturbed in SDs: physical well-being, psychological well-being, self-care and independent functioning, occupational functioning, interpersonal functioning, personal fulfillment, and overall quality of life. No differences between patients and normal healthy subjects where found in the components social support, community and services support or spiritual fulfillment. Patients suffering from nonorganic SDs had significantly worse scores in physical and psychological well-being and overall quality of life than those with organic SDs. Patients with both SDs and additional diagnoses of affective disorders had more profoundly reduced HRQoL than those with anxiety disorders. Follow-up of 51 patients (31 with nonorganic SDs and 20 with organic SDs) one year after sleep laboratory investigation and subsequent treatment found significantly improved HRQoL compared with pre treatment. Moreover, patients diagnosed and treated in the sleep laboratory showed lower re-hospitalization rats. PMID- 12778779 TI - [Evaluation of a new insulating system for infusion solutions in preclinical trauma therapy: a prospective, randomized study]. AB - OBJECTIVE: Infusion of cold fluids in a patient leads to a reduction of core temperature and subsequently worsens hypothermia. We evaluated the efficacy of a newly developed self-warming insulation device for use in pre-hospital rescue. METHODS: We studied 50 trauma patients with a rescue time of more than one hour. They were randomly assigned to either infusions taken directly from a warming box in the ambulance (Group A, n = 25) or infusions taken from the warming box and packed in an insulation device (Group B, n = 25). We recorded ambient temperatures, infusion temperatures in five-minute-steps and transport duration of the infusions from the ambulance to the site of accident. RESULTS: Ambient temperatures and transport duration did not differ significantly between both groups. In Group A the infusion temperature decreased from 36.0 +/- 6.4 degrees C to 19.8 +/- 6.8 degrees C during the transport from the ambulance to the site of accident. In Group B infusion temperature decreased only about 1 degree C. In Group A the temperature of the infusion continued to decrease until the end of measurements. In contrast in Group B the infusion temperature even increased by 0.5 degree C over the measurement period. These differences between the two groups were statistically significant. CONCLUSIONS: Our data show that even pre warmed infusions from a warming box cool down considerably before they can be given to the patient. A self-warming insulation device can stabilize infusion temperature even under extreme conditions of prehospital trauma care. PMID- 12778780 TI - First documented case of human granulocytic ehrlichiosis in Austria. AB - We report the first documented case of human granulocytic ehrlichiosis (HGE) in Austria. The infection was acquired near Arzl in the surroundings of Innsbruck in northern Tyrol. Except for a biphasic course of illness, presentation in this 33 year-old female patient was comparable to clinical findings observed in other European adults with high fever, transient thrombocytopenia, elevated levels of CRP and LDH, arthralgias, myalgias, fatigue and subfebrile temperature. Flu-like symptoms started about seven days after a tick bite and lasted for ten days, followed by an asymptomatic interval of three days and an acute onset of fever up to 39.5 degrees C on day 20. On admission, the patient showed high antibody titres against Anaplasma phagocytophilum (IgG 1:1024, IgM 1:640); six weeks later the IgG-titre had risen to 1:2048, and IgM-levels had fallen below 1:40. The demonstration of anti-platelet antibodies in acute-phase serum was noteworthy. We conclude that also in Austria HGE should be considered in patients with febrile thrombocytopenia, especially when the medical history reveals recent tick exposure. PMID- 12778781 TI - [Development of squamous epithelial carcinoma on the lower leg in long-term verrucous lichen planus]. AB - Lichen ruber planus and squamous cell carcinoma are common diseases of the skin. Several publications have dealt with the association between these two types of dermatosis. Extensive trials have shown the potentially premalignant nature of lichen planus of the oral mucosa. However, no study conducted so far was able to show that lichen planus of the skin increases the risk for squamous cell carcinoma. We report a 65-year-old man who developed a squamous cell carcinoma in a long standing verrucous lichen planus in the lower leg. PMID- 12778782 TI - "Philosophy" in medicine. PMID- 12778783 TI - [Immediate maxillary reconstruction after tumor excision by using lined buccal fat-pad flap with bone graft technique]. AB - OBJECTIVE: To evaluate the role of buccal-fat-pad flap in the lined coverage of the maxillary bone graft for the maxillary reconstruction after the tumor excision. METHODS: Based on the cadaver anatomical observation of the buccal-fat pad, 38 patients with the maxillary tumors were undertaking the treatment in this paper. Under general anesthesia, the buccal-fat-pad flap was elevated and transferred to the anterior or nasal cavities as a line of the sinus after the tumor was immediately excised through an intro-oral or external approach. The bone from the ribs was then grafted in the place for the maxillary support. RESULTS: After the anatomical evaluation, 38 patients were carried out the operation successfully. All of the grafted bones were well covered without bone exposure. The aesthetic appearance was achieved. CONCLUSION: Bone graft lined by the pedicled buccal fat pad flap may be a safe and good method for reconstruction of the maxilla with a lower rate of re-absorption and sequestration. PMID- 12778784 TI - [Intro-extro cranial approach treat serious post fronto-orbital fracture deformities]. AB - OBJECTIVE: To explore the method of treating serious secondary fronto-orbital fracture deformities through intro-extra cranial approach. METHODS: The fronto orbital fracture was divided into two types according to whether there were any large scale fronto-orbital bone defects: type I: Large scale fronto-orbital bone defect; type II: Concave fronto-orbital fracture deformity without large scale bone defect. Both types were treated through intro-extra cranial approach to expose the fracture site. For type I deformity, the bone defects were repaired and reconstructed with outer table of cranial bone and artificial bone. For type II, the deformity was repaired by osteotomy, bone reposition and internal rigid fixation. RESULTS: 18 cases were treated from June 1998 to October 2000, include type I, 12 cases, and type II, 6 cases. All the patients recovered well and the post-operative appearance were greatly improved. CONCLUSIONS: Intro-extra cranial approach can expose the fractured site better than the simple extrocranio approach, and make the operation more easily done. Combined with the technique of cranio maxillo facial surgery, the treatment can be more complete and the results can be more satisfactory. PMID- 12778786 TI - [The neurovascular anatomy and its clinical implication of the rectus femoris muscle]. AB - OBJECTIVE: The objective of this anatomic study was to investigate the intramuscular neurovascular configuration and to evaluate whether the muscle could be split into two functional units in transplantation. METHODS: Ten fresh cadavers and ten preserved cadavers were used in the study. A mixture of lead oxide, gelatin and water was injected to the femoral artery of the fresh cadaver. The rectus femoris muscle with its neurovascular pedicles was dissected and radiographed. RESULTS: Three vascular patterns of the rectus femoris muscle were found in the 40 cadaver legs. The muscle received its blood supply through a single vascular pedicle (12.5%), or a dominant pedicle with 1-2 ramified (80%), or two dominant vascular pedicles (7.5%). CONCLUSIONS: The study provided a detailed description on the intramuscular neurovascular territories of the rectus femoris muscle. Based on the neurovascular supply of the muscle, it is possible to subdivide the muscle into two functional units for segmental muscle transfer. PMID- 12778785 TI - [The effect of insulin-like growth factor 1 on the fusion of cranial suture]. AB - OBJECTIVE: To evaluate the effect of insulin-like growth factor 1 for the bone induction and the regulation for the fusion of the sagittal cranial sutures. METHODS: The cells, derived from cranial sutures in the newborn SD rats and the sagittal suture from the mice, were cultured with a serum-free medium and treated with and without insulin-like growth factor 1. The osteoblast phetotypes (osteocalcin, alkaline, osteoponcin and type-1 collagen) were measured with the RT-PCR and ELISA, and the explanted sagittal sutures were then evaluated under light microscopy. RESULTS: The cells, treated with the insulin-like growth factor 1, significantly produced more osteocalcin, alkaline, osteoponcin and type-1 collagen than those without insulin-like growth factor 1. The fusion of the sagittal suture explants will delay till to 30 days when it was not treated with IGF1. However, in the group with IGF1 the fusion was observed to start in 8 days, and a small amount of the sagittal suture fusion was found at the 20th day while a large amount was at the 30th day. CONCLUSION: The IGF1 has a direct effect on the fusion of cranial suture due to enhancing bone induction of cranial suture cell. PMID- 12778787 TI - [Labia minora skin flap vaginoplasty using tissue expansion]. AB - OBJECTIVE: This is to introduce a method for vagina reconstruction using the expanded labia minora flap. METHODS: Two tissue expanders were implanted in the labia minora bilaterally and expanded slowly over 4 weeks. In the operation, the expanded labial tissue was advanced as a bipedicle flap to line the reconstructed vagina. Five patients were treated with this method. Postoperative stent placement and dilation resulted in a vaginal canal exceeding 8 cm in depth. RESULTS: During the follow-up of 6 months to 2 years, four of the five patients got married. The vulva exhibited almost indistinguishable appearance. The reconstructed vagina had sensory and secretary functions. Its morphology and depth well meet the physiological demand. CONCLUSIONS: The modified method of tissue expansion vaginoplasty using the labia minora bipedicle flap is a good option for vagina surgery. The reconstructed vagina possesses the anatomical and physiological resemblance. PMID- 12778788 TI - [Systemic influence after an island flap with venous congestion-reperfusion injury]. AB - OBJECTIVE: To investigate the systemic influence after an island flap with venous congestion-reperfusion. METHODS: An island flap was formed in a Rat model. The vein in the pedicle was clamped for 2 hours, 6 hours and 10 hours and released. The ear microcirculation, levels of TNF alpha and IL-10 were measured, and the neutrophils sequestration in tissues were counted. The vascular structure of the lung and intestine were evaluated. RESULTS: There were significant changes in the ear microcirculation, neutrophils sequestration of the lung and the intestine in the 2 hours, 6 hours and 10 hours groups, and became more serious by the time increasing. The TNF alpha level reached in maximum at 1 hour after the reperfusion, while the IL-10 became to the lowest level at 3 hours after the reperfusion. However, the TNF alpha and IL-10 levels were significant high in the 6 hour and 10 hours groups, compared with the 2 hours group and the control, but there was no differences between the2 hours group and the control. CONCLUSIONS: Venous congestion-reperfusion of flap could injury the remote organs such as lung, intestinal etc. The injury could be more serious with the time increasing. PMID- 12778789 TI - [Immediately mammaplasty after resection of large breast fibroadenoma]. AB - OBJECTIVE: To explore an ideal technique for remodeling the breast after resection of large breast fibroadenoma. METHODS: Based on the principle of breast reduction, an operation plan was designed. The new locations of the nipple, and the areola and the area of the breast skin to be resected were marked. The preoperative-marked skin, epidermis, and the whole breast tumor were resected routinely. Then breast remodeling followed according to augmentation mammoplasty. Seven patients underwent this operation. RESULTS: The operative results were satisfactory in terms of multiple parameters during the follow-up period. CONCLUSIONS: This operation can effectively resect the breast tumor and remodel the breast simultaneously. PMID- 12778790 TI - [Breast ptosis correction with a knitted polypropylene mesh]. AB - OBJECTIVE: To evaluate a knitted polypropylene mesh used for mammapexy in correcting the breast ptosis. METHODS: Twenty-one patients with the mild or moderate breast ptosis were undergoing the mammapexy with a knitted polypropylene mesh. The operation was performed through the periareolar incision and a monofilament knitted polypropylene mesh was subcutaneously implanted on the surface of the gland and fixed upward. RESULTS: All of the patients were satisfactory of the appearance with the follow-ups from 2 to 16 months. No secondary ptosis, hypertrophic scars and foreign body reactions were found. CONCLUSION: The above-mentioned technique could be a good, safe and reliable method for correcting breast ptosis. PMID- 12778791 TI - [Analysis of the Millard method for unilateral cleft lip repair]. AB - OBJECTIVE: To analyze and describe the advantages and disadvantages of the Millard repair in the unilateral cleft lip (UCL). METHODS: In 30 patients with UCL undergoing the cleft lip repair with the Millard I or II method, the vermilion was repaired by a modified method with a triangle flap, while the alar cartilage reposition was performed. RESULTS: Postoperative follow-up of 6-12 months revealed the good results with invisible scar, good preservation of philtrum dimple and column, full vermilion and lengthened columella, good alar cartilage reposition. CONCLUSION: The Millard method is good for UCL repair. The modified technique with a triangle flap on the vermilion edge can overcome Millard's disadvantages. The anatomic reposition of the affected alar cartilage by blunt dissection at the first stage is suitable for the oriental. PMID- 12778792 TI - [Using scar tissue flap to repair secondary nasal deformities of unilateral cleft lip]. AB - OBJECTIVE: To introduce the clinical application of the upper lip scar tissue flap in correcting secondary nasal deformities of unilateral cleft lip. METHODS: Two types of the upper lip scar tissue flap were developed in correcting nasal deformities with Millard's technique. RESULTS: The procedure was performed on 51 patients. All the scar tissue flaps survived completely. The deformities were better corrected than before. CONCLUSION: The scar tissue flap has adequate blood supply. This procedure is routinely combined with other commonly used techniques to correct secondary deformities of cleft lip. It is particularly applicable for repairing the nasal deformity when usable tissue is not enough. PMID- 12778793 TI - [Autologous buccal mucosal graft for urethral reconstruction]. AB - OBJECTIVE: To search for a new method for urethra reconstruction using autologous buccal mucosal graft while lacking of local skin. METHODS: Since 1998, a total of 25 patients with complex hypospadias have been treated using buccal mucosal grafts for urethral reconstruction. The reconstructed urethra was anastomosed with the meatus half year later. RESULTS: All the reconstructed urethra survived without contracture or stricture except one infection, which healed with no adverse consequence. CONCLUSION: The key points for operation success is rich capillary network, thick epidermis and thin lamina propria of the buccal mucosa. Buccal mucosa is an excellent tissue for urethral reconstruction. PMID- 12778794 TI - [Local application of bFGF and sucralfate during continuous tissue expansion]. AB - OBJECTIVE: To investigate the effect of local application of bFGF and sucralfate during continuous tissue expansion (CTE). METHODS: CTE combined with local administration of bFGF and sucralfate was used in twelve patients with scar and nasal tip defects. Twenty three expanders were placed in the subcutaneous pockets through intralesion short incisions. Continuous expansion began at 1-3 days after expander implantation. The histomorphological changes and epidermal cell proliferation were observed. The clinical results were investigated. RESULTS: The average inflation time was 8.9 days. The average interval of the two operations was 13.5 days. The average hospitalization was 28.4 days. The average immediate stretch-back rate of the expanded skin was 25.7%. The clinical results were satisfactory without any complications. Histological examinations showed that the epidermal, granular and spinous layer became thicker. The basal cells increased significantly. The dermis thinned slightly and the collagen fibers became thicker. The elastic fiber regenerated significantly. Fibroblast and capillary density increased obviously. The immunohistochemistry analysis showed that the proliferation of epidemic basal cells was significant postoperatively. CONCLUSION: Local application of exogenous bFGF and sucralfate during CTE was feasible in patients. It could accelerate tissue expansion and improve the quality of expanded skin flap. PMID- 12778795 TI - [The expression of EST and ER in hemangioma with its clinical value]. AB - OBJECTIVE: To study the relationship between estrogen and the development of hemangioma. METHODS: The expression of EST and ER in samples from the thirty eight cases of hemangioma and six cases of normal control group was examined with the immunohistochemical steptavidin peroxidase conjugated method (SP method). RESULTS: The EST in capillary hemangioma expressed significantly higher than in the cavernous hemangioma, the racemose hemangioma or the control group. Although the EST in cavernous hemangioma and racemose hemangioma also expressed higher than in the control, there are no statistical differences among them. The ER only expressed in some cases in the capillary hemangioma group. No sexual difference was shown in the expressions of the EST and the ER. CONCLUSION: This study shows that there may be a relationship existed between the estrogen and the capillary hemangioma. It may indicate that some capillary hemangioma may be possibly treated by the drugs. PMID- 12778796 TI - [Fabrication of a blood vessel scaffold with a combined polymer for tissue engineering]. AB - OBJECTIVE: To investigate the possibility to fabricate a blood vessel scaffold with a combined polymer for tissue engineering. METHODS: A blood vessel scaffold was designed with a combined polymer composed of rabbit vascular smooth muscle cells(VSMCs), collagen and a non-spinning fabric mesh of polyglycolic acid (PGA). VSMCs were implanted into collagen gel and their growth was observed. The mixed solution of VSMCs and collagen was dropped into the tubular scaffold, followed by 7-day culturing. RESULTS: VSMCs formed many prominences after culturing in gelatinous collagen for 3-4 hours. With cells extending, some cells became shuttle- or spindle-shaped. After VSMCs-collagen complex was implanted into the PGA mesh, most of VSMCs remained in the pore of PGA mesh with the formation of gelation. VSMCs could adhere to and grow on the PGA fiber. CONCLUSION: The non spinning PGA porous biodegradable material coated with collagen is a good carrier for VSMCs to adhere and grow. PMID- 12778797 TI - [The expression and significance of structural proteins, VEGF and Ang-1 in cavernous venous malformations of the body surface]. AB - OBJECTIVE: To study the expression and significance of structural proteins, VEGF and Ang-1 in cavernous venous malformations of the body surface. METHODS: Tissue samples came from 25 cases of cavernous venous malformations, 12 cases of normal moderate veins and 12 cases of normal small veins. Envision immunohistochemical stain was used to investigate the expression of IV collagen, fibronectin, laminin, VEGF and Ang-1. The results were analyzed semi-quantitatively. RESULTS: The distribution of structural proteins in cavernous venous malformations is similar to moderate and small veins, but the expression in venous malformations is less obviously. VEGF expression in cavernous venous malformations and small veins is stronger obviously than moderate veins. Ang-1 expression in small veins is stronger remarkably than cavernous venous malformations and moderate veins. CONCLUSION: The abnormal expression of structural proteins may be an important factor in etiopathology and progress of cavernous venous malformations. There is disturbance of blood vessel remodelling in the sinusoid of cavernous venous malformations, with which the less expression of Ang-1 may be related. PMID- 12778798 TI - [Experimental study on effects of endothelin in the proliferation and collagen synthesis of human scar-derived fibroblasts]. AB - OBJECTIVE: To investigate the role of endothelin (ET) in the proliferation and collagen synthesis of human scar-derived fibroblasts and the modulation of its antagonists such as nitric oxide (NO), tetrandrine (Tet). METHODS: With the cultured fibroblasts from the scarring tissue, the cell proliferation was determined by [3H]-TdR incorporation, while the collagen synthesis was evaluated by [3H]-proline incorporation. RESULTS: The ET-1 was significantly increasing the proliferation and collagen synthesis of human scar-derived fibroblasts. The values of [3H]-TdR absorption in the 2.5 ng/ml, 25 ng/ml and 100 ng/ml of ET-1 groups were 1.8 times, 4 times and 4.9 times more than in the control group, respectively (P < 0.01), while the values of the [3H]-proline incorporation were 1.1 times, 3.1 times and 3.8 times respectively (P < 0.01). The fibroblasts, treated with 50 micrograms/ml of S-nitroso-N-acetyl penicillamine(SNAP), were no detectable effect on the basal level of DNA synthesis, but produced decreasing effect on the [3H]-TdR absorption (the rate of inhibition was 22.89%, P < 0.05). It was found that the SNAP inhibited the [3H]-proline incorporation in cultured fibroblasts, but the rate of [3H]-proline incorporation induced by ET-1 was unaltered. The Tet with 3 micrograms/ml, in which does not inhibit the basal level of DNA synthesis, was significantly decreasing the collagen synthesis and decreasing the ET-mediated DNA synthesis (the rate of inhibition was 33.21% (P < 0.01). CONCLUSION: These results indicate that the ET can obviously increase the proliferation and collagen synthesis of human scar-derived fibroblasts, but it can be partially antagonized by NO and Tet. PMID- 12778800 TI - [A review and expectation of my transmillenarian life]. PMID- 12778799 TI - [Influence of prostaglandin E2 on proliferation of melanocytes in full-thickness skin graft]. AB - OBJECTIVE: To investigate the influence of the prostaglandin E2 on the proliferation of the melanocytes in the full-thickness skin graft. METHODS: Sixty eight guinea-pigs were divided into experimental-1 group (skin graft), experimental-2 group (skin graft + diclofenac), and control groups. After the full-thickness skin graft, the dynamic changes of the prostaglandin E2 were measured and the proliferation of the melanocyte with its density was also evaluated by using histochemical and autoradiographic methods. RESULTS: In the experimental-1 group, the content of PGE2 was increasing in seven days after the operation, continued to the one month, and then returned to the base level. The labelling indices of 3H-MC-TdR of the group was also increasing postoperatively between the second day and the fourteenth day, and reach a second peak after one month, then came to the normal level. The density of the melanocytes was decreasing rapidly 3 days after the surgery, then began to increase and exceeded over the normal level 21 days after the operation. However, in the experimental-2 group, the content of PGE2 decreased in two days after the surgery, and then showed the inclination similar to the experimental-1 group with the different points in narrower range. The number of melanocytes labelled by 3H-TdR began to increase at the first day after the surgery, which appeared earlier than the experimental-1 group and was similar in the changing tendency with a less extent. The density of MC showed the similar tendency to the experimental-1 group in a narrower changing range with both of increasing and decreasing. The density of the MC was much lower in 21 after the operation than the experimental-1 group and normal control group. CONCLUSION: The increased PGE2 in the earlier stage of the skin grafting could enhance the inflammatory reaction to the tissue, as well as the melanocytes. It may stimulate the proliferation of the MC with the result of increasing their density. The use of the diclofenac might reduce the inflammation and suppress the proliferation of melanocytes, and result in the skin with light color due to decreasing the number of MC in the epidermis of the graft. PMID- 12778801 TI - [Proteins regulating neurotransmitter release of synaptic vesicles at nerve terminals]. AB - Neurotransmitter release of synaptic vesicle at nerve terminals is a complicated and elaborately regulated process, involving a cascade of protein-protein interactions. The SNARE core complex consisting of synaptobrevin/VAMP (a synaptic terminal protein), syntaxin and SNAP-25 (synaptic membrane-associated proteins), acts as the membrane fusion machinery and plays essential roles in synaptic vesicle exocytosis. This review will discuss proteins with potential roles in synaptic vesicle exocytosis by regulating assembly, disassembly and the function of SNARE complex, and sum up the molecular model of vesicle exocytosis. PMID- 12778803 TI - [Progress in the study of physiological function of sphingosine 1-phosphate]. AB - Sphingosine 1-phosphate, a bioactive sphingolipid produced from the metabolism of sphingomyelin, play important roles in diverse biological process, including cell proliferation, survival, cytoskeleton changes, migration, angiogenesis, wound healing and embryonic development. Here we review the role of sphingosine 1 phosphate in cell biological function regulation and signal transduction. PMID- 12778802 TI - [Capsaicin and its receptor--vanilloid receptor]. AB - The detection of painful stimuli occurs primarily at the peripheral terminals of specialized sensory neurons called nociceptors. These small--diameter neurons transmit this information to the central nervous system, ultimately eliciting a perception of pain or discomfort. The capsaicin (vanilloid) receptor, an excitatory ion channel expressed at nociceptors, plays an important role in transducing thermal and inflammatory pain. Mice lacking the VR1 gene have deficits in thermal- or inflammation-induced hyperalgesia, which confirms the involvement of this channel in pain sensation, especially the sensation of heat evoked pain. PMID- 12778804 TI - [Effects of insulin on the diseases of central nervous system]. AB - There are increasing experimental evidence and clinical data showing that insulin plays an important role in the CNS. Insulin can be found at high level in the brain of some animals. Insulin receptors and insulin-second messengers systems are present in neurons and glial cells. The pathogenesis of some neurological diseases is related with insulin level or insulin sensitivity. Insulin-like growth factors also have a modulatory role in neuronal function. Insulin and neurotrophic factors including insulin-like factors have generated considerable excitement for their potential as therapy for a wide variety of degenerative neurological disorders, for which there is currently no treatment. PMID- 12778805 TI - [Current progress of phenotypic modulation of vascular smooth muscle cells]. AB - Proliferation and migration of vascular smooth muscle cells were the common pathological feature of development and progression of atherosclerosis, hypertension and vascular restenosis. Phenotypic modulation of vascular smooth muscle cells was the basis of its proliferation and migration. So, understanding the mechanisms of phenotypic modulation of these cells may be helpful to prevent and treat the diseases mentioned above. This paper will review the influence factors, signal transduction and transcription factors involved in phenotypic modulation of vascular smooth muscle cells. PMID- 12778806 TI - [Alterations of G proteins in heart diseases]. AB - Guanine nucleotide binding proteins (G proteins), known as an important signal transduction molecular, can physiologically couple extracellular signals identified by G protein-coupled receptors (GPCRs) to a series of intracellular effector to cause changes of gene transcription and protein's structure and function. Members of G protein family including Gs, Gi/o, Gq/11, and G12/13 express in the myocardium. In the heart G protein-mediated signal transduction generates diverse effects on myocardiac contractility, heart rate, heart rhythm, and myocyte growth. This review focuses on alterations of G protein in cardiac hypertrophy, heart failure, acute myocardial ischemia, and arrhythmia to help us to understand the pathogenesis and pathophysiology in heart diseases. PMID- 12778807 TI - [The role of neuronal nicotinic acetylcholine receptors in the mechanisms of general anesthesia]. AB - Neuronal nicotinic acetylcholine receptors (nnAChRs) are members of the ligand gated ion channel superfamily, and widely expressed in the central and peripheral nervous systems with many subunits. NnAChRs have been represented novel targets for a wide variety of therapeutic agents based on their complex functions. It is known that both central and peripheral nnAChRs are sensitive to various types of general anesthetics, among those, barbiturates, ketamine, volatile and gaseous anesthetics depress nnAChRs at or below clinical concentrations. It is possible that inhibition of nnAChRs is one of factors involved in the mechanisms of general anesthesia. PMID- 12778808 TI - [Development and compensation of pancreatic beta cells]. PMID- 12778809 TI - [Astrocyte: a new star in pain research]. PMID- 12778810 TI - [Research progress in two-pore-domain K+ channels]. PMID- 12778811 TI - [A new member of histamine receptor family: H4 receptor]. PMID- 12778812 TI - [New ideas in the study of gene function]. PMID- 12778813 TI - [ABCG2/Bcrp1 transporter as phenotype markers and regulators of side population stem cells]. PMID- 12778814 TI - [The excitatory role of GABA during the early development of the central nervous system]. PMID- 12778815 TI - [Opioids and ischemic preconditioning]. PMID- 12778816 TI - [The relationship between iron metabolism in central nervous system and Parkinson's disease]. PMID- 12778817 TI - [The structure and function of RAGE]. PMID- 12778818 TI - [The prognostic relation of nm23-H1/NDPK-A with leukemia and lymphoma]. PMID- 12778819 TI - [Putative role of 5-lipoxygenase in central nerve system]. PMID- 12778820 TI - [RNA interference: one of physiologic mechanisms on RNA-mediated memory transfer]. PMID- 12778821 TI - [Cyclooxy genase-2 and tumor progression]. PMID- 12778822 TI - [The relation of human and mammalian fertilization and glycoproteins]. PMID- 12778823 TI - The state of the science of urinary incontinence. PMID- 12778824 TI - Interstitial cystitis: an updated overview. AB - Interstitial cystitis (IC) is a chronic inflammatory condition of the bladder wall, characterized by pelvic pain, urinary urgency, and frequency. Its etiology is unknown. There are an estimated 700,000 cases of IC in the United States. IC remains a diagnosis of exclusion. Although there are no uniformly effective therapies, many treatment options are currently available. Current research is focused on the cause, diagnosis, and treatment of IC. PMID- 12778825 TI - Me and my bladder, my bladder and me. AB - This article is intended to "put a face" on the disease interstitial cystitis (IC). It is a personal case study based on the experiences of the author. Being misdiagnosed and dismissed for years, her story is typical of the IC patient's struggle. If one IC patient is recognized and treated in a timely fashion, then those years of being misunderstood would have counted for something. PMID- 12778826 TI - The effects of a continence program on frail community-dwelling elderly persons. AB - Urinary incontinence among community-dwelling frail elderly is estimated to affect 6.23 million people at an annual cost of over $7 billion. Overwhelmed caregivers may seek institutional placement of their incontinent family member. Results from this study of 78 subjects demonstrate that even in physically and cognitively impaired persons a behavioral treatment program can reduce incontinence, lower costs, and relieve caregiver burden. PMID- 12778827 TI - Urinary dysfunction and urodynamics in the elderly. AB - As the elderly population continues to grow, it is important to consider some of the normal changes inherent with the aging process, as well as to understand common urologic disorders that may occur in this population. Urodynamic clinicians should be aware of some special considerations when working with this group of individuals. The purpose of this article is to describe common urologic conditions in the elderly and correlate these conditions with typical urodynamic findings. PMID- 12778828 TI - A pilot study comparing two methods of intermittent catheterization: limitations and challenges. AB - Given a complex research design, a pilot study was conducted to compare a closed catheter system for intermittent catheterization to the traditional open system. Based on 33 urine samples for 11 subjects, the closed system yielded fewer positive cultures. Efforts to obtain this preliminary data proved beneficial in identifying unanticipated design limitations that will guide modifications of the main study, which will include multiple centers. PMID- 12778829 TI - An evaluation of a specialist nursing role for men with prostate cancer. AB - The role of a specialist prostate nurse in the care of men with newly diagnosed prostate cancer has not been well documented. Therefore, the needs of men diagnosed with prostate cancer were individually assessed in this qualitative study. The nurse was an acceptable source of information, with time to listen and respond to the men's physical and psychosocial needs at diagnosis and throughout the continuum of care. PMID- 12778830 TI - Demystifying the clinical nursing research process. PMID- 12778831 TI - Aminocaproic acid (Amicar): potent antifibrolytic agent for treating hematuria. PMID- 12778832 TI - Herbs, amino acids, and female libido. PMID- 12778833 TI - Getting ready for certification: obstructive uropathies. PMID- 12778834 TI - Osteoporosis. Part IV -- rapid review of drug therapies (A to Z) for preventing male osteoporosis/fractures.. AB - Numerous drug therapies exist for women in terms of prevention and treatment of osteoporosis and fractures from this same condition. However, the data for effective drug therapies are limited in men, but are beginning to accumulate rapidly. Most of the available drug treatments for women should be reviewed with men when considering osteoporosis drug prevention for males. Numerous potential possibilities of effective drug treatment for males osteoporosis exist and the number of future therapies seems limitless, but weighing the potential benefits and detriments of each currently available treatment is time consuming. PMID- 12778835 TI - Pharmacogenomics of psychiatric drug treatment. AB - It is the goal of pharmacogenomics in psychiatry to establish predictive relationships between polymorphisms of candidate genes and therapeutic response to drug treatment. Polymorphisms of candidate genes related to drug mechanisms and pathophysiology of illness and defined clinical phenotype are the foundations for pharmacogenomic studies. Pharmacogenomic studies of antipsychotic response have focused on polymorphisms of genes for dopamine and serotonin receptors with most positive results reported for polymorphisms of genes of the 5HT2a and 5HT2c serotonin receptor subtypes. Although the goal of establishing individualized medicine predicated on an individual patient's genetic code has yet to be achieved, the fundamentals are now in place for second-generation investigation and more application to health care. PMID- 12778836 TI - Biological predictors of treatment response in affective illness. AB - Attempts to identify biological response predictors generally have met with limited success, particularly where the goal is to develop clinically useful indices. This article reviews the biological approaches to predicting treatment response, beginning with neuroendocrine studies and electroencephalogram analysis and concluding with structural and functional neuroimaging. The article describes the designs of typical studies to aid in interpreting their results and concludes by addressing some of the problems and limitations associated with these approaches and suggesting future directions for this research. PMID- 12778837 TI - Predictors of response to antidepressants general principles and clinical implications. AB - What variables will predict who should be treated with antidepressants and who should not? If someone should be treated, what variables will predict which treatment is most likely to work? If patients are treated, what will predict who will respond and who will not? Clinicians have asked these questions ever since researchers established that antidepressants were superior to placebo in the treatment of depression. Innumerable reports claim either biologic or psychosocial predictor variables of response, and every few years yet another review of predictors is published in the literature. This article reviews methodologic issues that have been neglected in the pursuit of predictors to antidepressant response and proposes improved requirements for predictor research that would impact clinical practice. PMID- 12778838 TI - Predictors of response in depression. AB - Conflicting or sparse data on predictors of treatment response in depression have resulted in lack of clear guidelines in choosing antidepressant treatment. Critical to treatment outcome are accurate diagnosis and adequate treatment. Other data easy to obtain, such as age, gender, and marital status, have failed to be consistent predictors; more elaborate studies, such as receptor analysis or neuroimaging, are not yet accessible to most clinicians or economically feasible; however, they offer hope for the future, when more biologically based diagnostic distinctions may come to guide treatment choices. PMID- 12778839 TI - Treatment of schizophrenia: preventing the progression of disease. AB - Atypical antipsychotic drugs offer a number of advantages compared with typical antipsychotic drugs during the acute treatment of psychotic episodes and maintenance treatment to prevent psychotic relapses in patients with schizophrenia. These advantages include superior efficacy, especially for negative symptoms and reduced extrapyramidal adverse effects. Clinicians, however, need to be vigilant regarding new adverse effects that may be more strongly associated with atypical than typical drugs, especially those related to weight gain. Despite increased pill costs for atypical antipsychotic drugs, the overall costs of providing care to patients with schizophrenia using these drugs do not appear to be substantially higher. PMID- 12778840 TI - Outcome predictors in substance use disorders. AB - Given the heterogeneous nature of substance abuse, it is notable that several predictors of response are independent of the primary drug of abuse or the treatment setting [208]. Although the strength of the relationship of predictor to outcome varies, the following factors have been identified consistently: severity of dependence or withdrawal; psychiatric comorbidity; substance-related problems; motivation (abstinence commitment); length of treatment; negative affective states; cognitive factors; personality traits and disorders; coping skills; multiple substance abuse; contingency contracting or coercion; genetic factors; sleep architecture; urges and craving; self-efficacy; and economic and social factors. Although it is well known that severity of dependence (including polysubstance abuse), serious psychiatric comorbidity, and social problems are associated with poor treatment response, only recently has research examined the efficacy of intervention strategies that specifically address these problems. Adequate treatment of psychiatric comorbidity and improvement in social, economic, and family functioning lead to better treatment outcomes. The development of specific techniques to enhance self-efficacy, motivation, coping skills, and functioning in the community are concrete examples of how the identification of factors associated with positive outcomes has led to the development of new treatments. Despite significant accomplishments, the field is left with many unanswered questions. Although several biologic markers, such as neuroendocrine response and sleep architecture, show promise as outcome predictors, it is not known whether these are critical factors in the initiation of substance use or its progression to dependence. Determining whether biologic markers are epiphenomena reflecting the amount and duration of substance abuse or are fundamental to the pathophysiology of dependence is a matter of urgent concern. With some exceptions, identification of biologic predictors has not led to innovative therapies. One of these exceptions is the development of naltrexone for the treatment of alcoholism, which was based in a solid theoretical rationale and followed by hypothesis-driven experiments. Similar opportunities should emerge from current basic science and clinical research. The application of pharmacogenetic techniques to the field of addiction also holds great promise. As future studies are undertaken, researchers and clinicians must be mindful that differences in outcome predictors across drugs of abuse and treatments may emerge as subgroups of individuals with addictive disorders and new therapies are identified. There is already evidence that early onset alcoholism is associated with poor response under some circumstances, yet may be a predictor of response to targeted pharmacotherapy with ondansetron [64, 112]. As the ability to subtype disorders based on meaningful biologic differences grows, it is anticipated that several relevant outcome predictors that are specific for pharmacotherapy will emerge. PMID- 12778841 TI - Predictors of response in anxiety disorders. AB - Anxiety disorder variables such as duration, severity of illness, and comorbidity with other anxiety or mood disorders appear to identify individuals who are at the greatest risk of treatment nonresponse. Conversely, in accord with clinical experience, shorter periods of illness, less severe illness, being treatment naive, and the absence of comorbidity tend to identify patients who are likely to respond robustly to medication management. Symptom clusters in OCD and PTSD are promising as a means of stratifying those more likely to respond to standard pharmacologic treatment. The presence of hoarding or sexual obsessions seems to presage poorer response in OCD, while the presence of dissociative symptoms in PTSD has been linked to high nonspecific treatment response rates to placebo. Genotyping individuals with respect to genes that are thought to have an important role in the underlying disease process, such as the work with the 5HTTL PR allele, is exciting and is perhaps the first glimmer of using genotyping to identify treatment strategies or to predict the likelihood or speed of response. The use of neuroimaging as a means of identifying individuals who may respond favorably to pharmacologic or neurosurgical intervention is still in its infancy. As a strategy, it may help combine symptom severity and response variables into a clear neurobiologic vulnerability model of illness. In the future, it may be possible to identify specific treatment interventions for specific patterns of abnormal metabolic rates in certain areas of the brain. However, it should be emphasized that such an approach has not been empirically demonstrated in a rigorous experimental context at this time. PMID- 12778842 TI - Predictors of response to treatment in children and adolescents with mood disorders. AB - Depression and bipolar disorder are frequently chronic disorders, with onset often beginning in childhood. Mood disorders are becoming more recognized in children and adolescents, and treatment of these disorders has received much attention, particularly in the past 10 years. Recent studies have demonstrated efficacy of antidepressant medications (particularly SSRIs) and specific psychotherapies (primarily CBT). Rates of remission (little or no symptoms) in these studies, however, have remained quite low (35% to 40% in most acute studies). Furthermore, recurrence is common in this population, and affects 40% to 50%. Early onset mood disorders are also associated with increased risk of developing other psychiatric disorders, substance abuse, and suicide, and having poor academic, work, and social functioning. The lifelong implications are serious. Identifying factors that may predict response to treatment, both in general and to specific treatments, may lead to improved outcomes for these patients. Unfortunately, studies have typically been inconsistent. Most studies do not identify demographic variables as predictive of outcome, although older age has been associated with poor prognosis in several studies. Psychosocial factors have yielded some results, particularly with regard to family environments. Generally, intact families with positive interaction styles and less dysfunction have been associated with better outcomes. Psychiatric disorders among parents not only predicts the development of the disorder, but is also associated with poorer prognosis. Finally, several clinical factors have been linked to poorer outcome in children and adolescents with mood disorders. More frequent episodes, increased severity (particularly suicidality and psychosis), and comorbid disorders are likely to lead to fewer recoveries, longer episodes, and increased rate of recurrence. Recent attention has focused on mediators and moderators of outcomes to treatment. In general, the theory is that enumerable factors contribute to the course of an individual's mood disorder, but that by identifying some of the variables that have more impact may allow for more specific or modified treatments to improve outcome. Many of the predictive factors explored in this article are examples of mediators and moderators that affect outcome. Each one alone may not provide definitive answers for predicting response to treatment, but each must be taken into account at the outset of treatment. It is clear that treatments must be individualized for each patient. Furthermore, selecting only one treatment exclusively for patients may hinder progress. The first step is to attempt to identify some of the underlying causes and the consequences of the disorder itself (i.e., decreased social interaction). The next step in successful treatment is to address both the causes and consequences of the disorder, through medication, psychotherapy, skills training, family intervention, or any other methods needed to assist the child to begin functioning better in all domains (social, academic, work, family, and so forth). Such a biopsychosocial approach to treatment of these disorders will likely improve overall outcome. PMID- 12778843 TI - Background and rationale for the sequenced treatment alternatives to relieve depression (STAR*D) study. AB - Sequenced Treatment Alternatives to Relieve Depression (STAR*D) attempts to fill in major clinical information gaps and to evaluate the theoretical principles and clinical beliefs that currently guide pharmacotherapy of major depressive disorder. The study is conducted in representative participant groups and settings using clinical management tools that easily can be applied in daily practice. Outcomes include clinical outcomes and health care utilization and cost estimates. Research findings should be immediately applicable to, and easily implemented in, the daily primary and specialty care practices. This article provides the overall rationale for STAR*D and details the rationale for key design, measurement, and analytic features of the study. PMID- 12778844 TI - Treatment of bipolar depression: current status, continued challenges, and the STEP-BD approach. AB - Even though at least 10% (if not 20%) of those who experience a first lifetime episode of depression will subsequently develop bipolar disorder, the alliance of academic and industry research agendas that leads to developing and testing new antidepressants has failed to produce a sufficient knowledge base. It is therefore impossible to apply a truly empirical approach to guide the treatment of people with bipolar depression. Consequently, there are holes in contemporary evidenced-based practice guidelines large enough to drive a truck through; furthermore, there are some recommendations that have no factual basis other than expert opinion. However, with new research emerging on lamotrigine and olanzapine, in addition to the pending results of larger studies supported by the National Institute of Mental Health and the Stanley Foundation, there is evidence that some progress is being made. PMID- 12778845 TI - [Chromosome abnormalities in hyperparathyroidism: utility of comparative genomic hybridization in the study of parathyroid hyperplasias]. AB - Genetic abnormalities responsible for primary (pHPT) and secondary hyperparathyroidism (sHPT) are not well described, especially those underlying the autonomous and refractory behaviour of glands from uremic patients with glandular hyperplasia and nodular growth. Comparative Genomic Hybridization (CGH) is a molecular cytogenetic technique based on a double-color in situ fluorescent analysis, allowing a global description of gains and losses of genomic material. It is a useful tool that localizes unstable genetic areas whose alteration could modify the expression of one or several genes related to the pathology in study. Results on primary hyperparathyroidism adenomas have shown a series of genetic changes correlating with areas where genes related to pHPT are located, such as MEN1 and cyclin D1. A large number of chromosomal aberrations in glands from patients with secondary hyperparathyroidism have also been found, and although some of them are common with those described for primary hyperparathyroidism, most of them are located in different areas or in a different proportion. These results confirm that although severe sHPT hyperplasias can evolve into neoplasias similar to pHPT adenomas, both parathyroid alterations must be considered, from a genetic point of view, as unrelated. PMID- 12778846 TI - [Regulation of the calcium-sensing receptor. Influence of secondary hyperparathyroidism]. AB - The parathyroid glands have a great sensitivity to small changes in the extracellular ionic calcium. The calcium-sensing receptor (CaR) is a G protein coupled receptor that responds to extracellular ionic calcium changes activating several intracellular signalling systems (phospholipases C, A2 and D) finally inhibiting the PTH secretion. In addition to calcium, there are some other agonists and modulators such as the Mg2+, spermine, amyloid beta-peptides, a variety of aminoacids, especially aromatic aminoacids and ionic strength. In the uraemia, the sensitivity of the parathyroid glands to calcium is altered and higher values of calcium are necessary to suppress the PTH. In the secondary hyperparathyroidism the CaR expression is reduced. It has been found a negative correlation between cellular proliferation and the expression of the CaR in hyperplasic glands. Despite it is a calcium receptor, the expression of the CaR does not seem to be regulated by calcium and there is some controversy about the role of calcitriol regulating its expression. On the other hand, the phosphorous induces hyperplasia of the parathyroid gland increasing the cellular proliferation and a decrease of the CaR expression. PMID- 12778847 TI - [The PTH/PTHrP receptor: biological implications]. AB - Since its discovery in 1923, the parathyroid hormone (PTH), was thought to be the sole hormone capable of stimulating bone resorption, renal tubular calcium reabsorption, calcitriol synthesis, and urinary excretion of phosphate. However, in 1987, the PTHrP (PTH-related peptide), was demonstrated to share most of the biological actions of PTH through the activation of the same receptor. This receptor was cloned in 1992 and named PTH/PTHrP receptor or PTH-R1. Both, PTH and PTHrP bind with great affinity to PTH-R1 and stimulate a signal transduction system involving different G-proteins, phospholipase C, and adenylate cyclase. A third member of the PTH family, the TIP-39 (tuberoinfundibular peptide), binds and activates another PTH receptor (PTH-R2). There is evidence for other PTH receptors, a PTH-R3, probably specific for PTHrP in keratinocytes, kidney, placenta and a PTH-R4 specific for C-terminal PTH fragments. Activating mutations in the PTH-R1 gene cause Jansen type metaphyseal chondrodysplasia, whereas inactivating mutations are responsible for Blomstrand type rare chondrodysplasia and enchondromatosis. The renal and bone PTH-R1 expression is upregulated in vitamin D deficient rats and by endotoxin, interleukin-2, dexamethasone, T3, and TGF beta. On the contrary, PTH, PTHrP, angiotensin-II, IGF-1, PGE2, vitamin D, and chronic renal failure decrease its expression. In conclusions, the biological implications of the identification and cloning of different PTH receptors are at their beginning. The almost ubiquitous distribution of PTHrP and PTH-R1, the numerous PTHrP and PTH fragments, let us suppose the existence of other PTH related receptors, and a great complexity of the bone and mineral metabolism. PMID- 12778848 TI - [The growth plate in chronic renal insufficiency]. AB - Several alterations have been reported in the growth plate of young rats rendered uremic by subtotal nephrectomy, a widely used experimental model of growth failure secondary to renal insufficiency. In our lab's experience, uremia is associated with a markedly increased growth plate height which results from an elongation of the hypertrophic zone. These findings are not consistently observed in all studies, likely because of the different experimental conditions. Regardless of growth plate size, growth retardation induced by chronic renal failure is accompanied by an alteration of the dynamics of the growth plate with a decreased bone apposition rate at the metaphyseal end of growth cartilage and slower production and progression of chondrocytes from the resting zone up to the most distal hypertrophic zone adjacent to bone. These abnormal dynamics are associated with an irregular bone-cartilage interface and a disturbed process of chondrocyte maturation which becomes evident by a morphological criteria and by depressed expression of markers of chondrocyte maturation such as collagen X. The microscopic findings also suggest a disturbed process of capillary invasion, which precedes formation of new osseous tissue in the primary spongiosa, although the levels of vascular endothelial growth factor, as measured by immunohistochemistry, have been reported to be similar in the growth plate of uremic and control rats. The meaning of these findings in the pathogenesis of growth impairment secondary to chronic renal failure remains to be determined. PMID- 12778849 TI - [Effect of growth hormone and calcitriol on the growth plate in uremic rats]. AB - This study analyzed the modifications induced by growth hormone (GH) and/or calcitriol treatments in the growth plate of growth retarded uremic rats. Four groups of 5/6 nephrectomized rats were studied: untreated (U), treated with GH (U + GH), treated with calcitriol (U + D), treated with GH and calcitriol (U + GH + D). Treatments were given intraperitoneally during the second week of renal failure. Uremic groups were compared with sham-operated rats fed ad libitum (C) or pair-fed with U (CP). In comparison with C and CP, histomorphometric analysis of tibial proximal ends of U group showed decreased bone formation, as estimated by osseous front advance (OFA), elongation of growth cartilage and its hypertrophic zone, and decreased size of most distal chondrocytes. The U + D group tended to normalize growth cartilage height, and that of its hypertrophic zone, as well as the size of chondrocytes. In U + GH group OFA improved and chondrocyte size became normal, but growth cartilage remained elongated. Similar results were found in the U + GH + D group. These findings indicate that, in chronic renal insufficiency, the beneficial effect of GH on growth is not associated with normalization of growth cartilage morphology and that calcitriol facilitates chondrocyte maturation. When given together the effect of GH prevails. PMID- 12778850 TI - [Effect of desferrioxamine and deferiprone on osteocalcin secretion in osteoblast type cells]. AB - Desferrioxamine and deferiprone are both metal-chelating drugs often used in aluminum-overloaded dialysis patients. In these patients, desferrioxamine produces an improvement on bone mineralisation without a relevant decrease in bone aluminum. Thus, desferrioxamine might have a direct effect on bone cells. The aim of this study was to assess the effect of desferrioxamine and deferiprone on 1,25(OH)2D3-stimulated osteocalcin secretion in osteoblast--like cells. The study was carried out in MG-63 cell cultures. Cells were seeded at a density of 15,000 cel/cm2 and grown to confluence for 72 hours in DMEM supplemented with 10% FCS. The medium was then replaced by another medium containing 1% BSA, 10(-9) M 1,25(OH)2D3 and desferrioxamine 5, 10, 20, 40, 60, 80 microM or deferiprone 15, 30, 60, 120, 180, 240 microM. Tris-HCl at pH 7.4 was used as control. After 48 hours, supernatants were collected for the measurement of secreted osteocalcin. Desferrioxamine and deferiprone, at high doses (desferrioxamine: 60 microM, 80 microM; deferiprone: 180 microM, 240 microM), inhibited the 1,25(OH)2D3-induced osteocalcin secretion. On the contrary, at lower doses (desferrioxamine 5 microM; deferiprone 15 microM) stimulated the secretion. In summary, these results suggest that desferrioxamine and deferiprone exert a direct effect on bone cell metabolism that might be independent from their metal-chelating properties. PMID- 12778851 TI - [Influence of the vitamin D receptor gene polymorphism and 25-hydroxyvitamin D on arterial pressure in health individuals]. AB - The role of vitamin D in the regulation of blood pressure is unclear. There are no studies that relate Bsm I polymorphism with blood pressure. OBJECTIVE: To analyze if Bsm I polymorphism and 25-hydroxyvitamin D (25OHD3) influence blood pressure in healthy individuals with normal blood pressure. METHODS: Systolic (SBP) and diastolic (DBP) blood pressure, Body Mass Index (BMI), plasma creatinine, serum calcium, serum phosphorus, serum iPTH, serum 25OHD3 and Bsm I genotype were determined in 590 healthy individuals (260 men and 330 women). Data were analysed using a multiple linear regression model. SBP and DBP were defined as dependent variables and the rest of variables as independent. RESULTS: Gender was strongly associated with both SBP (beta: -12.01, p: 0.000) and DBP (beta: 4.78, p: 0.000). Therefore, a separate analysis was performed according to gender. In males, SBP was associated with BMI (beta: 0.83, p: 0.001), 25OHD3, (beta: 0.36, p: 0.000) and genotype (beta: -3.90, p: 0.002); and DBP with 25OHD 3 (beta: 0.16, p: 0.018) and age (beta: 0.28, p: 0.000). Differences of blood pressure among the three genotypes were explored by analysis of variance. SBP was higher in men with bb genotype than in the other genotypes (p: 0.007). In females, 25OHD3 and genotype were not associated with blood pressure. CONCLUSIONS: Healthy men with higher levels of vitamin D have higher levels of SBP and DBP. Moreover, men with bb genotype have the highest levels of SBP. Blood pressure levels in women are not influenced by vitamin D nor by Bsml genotype. Our data suggest a possible pathophysiological interaction between vitamin D and sex hormones in blood pressure control. PMID- 12778852 TI - [Effect of PTH, phosphate, and metabolic acidosis on the progression of renal insufficiency in the azotemic rat]. AB - In a previous study we have observed that NH4Cl-induced metabolic acidosis halted the progression of renal disease in azotemic rats with a high phosphate diet. We hypothesized that NH4Cl-induced metabolic acidosis may exert its protective effect by decreasing renal calcium content independent of serum levels of PTH and phosphate loading. To test this hypothesis we studied azotemic rats with very low phosphate diet or parathyroidectomy. Rats with low phosphate diet and parathyroidectomized rats developed renal failure after 5/6 nephrectomy, and in both groups the acid loading significantly decreased the progression of renal disease. Calcium renal content increased in both groups, even in rats with low phosphate diet, and this effect was also significantly decreased after an acid loading. Rats with acid loading developed greater hypertrophy of renal tissue than rats without acid loading. We conclude that NH4Cl-induced metabolic acidosis halted the progression of renal disease by decreasing calcium precipitation on renal tissue. Parathyroidectomy did not prevent progression of renal disease nor calcium precipitation, and a low phosphate diet in azotemic rats did not prevent increased calcium content on remnant renal tissue. PMID- 12778853 TI - [Evaluation of osteodystrophy parameters in a pre-dialysis unit]. AB - A cross-sectional study was developed with 100 of the first-time pre-dialysis patients visiting the Princesa University Hospital's Advanced Chronic Kidney Disease (ACKD) unit, with the aim of analysing various parameters of osteodystrophy at this time. Parameters evaluated were: age, gender, renal function, osteodystrophy serum parameters, comorbidity index (ICED) and the patients' origin to establish correlations between these parameters. Mean iPTH levels were higher irrespective of the patients' origin, and were significantly higher in men than in women, the former also having poorer renal function and higher comorbidity score. The mean levels of calcium, phosphorous, alkaline phosphatase and CO2 did not justify this rise in iPTH. Nutritional parameters NPNA and albumin were adequate in spite of ageing. At early stages of ESRD, iPTH could be elevated and ACKD units play an important part its early detection and subsequent treatment. PMID- 12778854 TI - [Biochemical and histological spectrum of renal osteodystrophy in Argentina]. AB - Between 1994-2001 we have performed 57 bone biopsies for diagnostic purposes in symptomatic CRF patients. We analyzed here 52 samples where the material was optimal for study, and divided them into 2 periods according to when the biopsy was performed: 1994-1996 and 1997-2001, to verify changes in the spectrum of renal osteodystrophy. Mean serum values were: serum calcium 9.9 +/- 1.8 mg/dl, phosphate 5.8 +/- 3.2 mg/dl, alkaline phosphatase 693.9 +/- 968.9 Ul/L, iPTH 562.0 +/- 598.5 pg/ml, serum aluminum 65.7 +/- 79.3 ug/L and bone aluminum 22.8 +/- 22.4 ug/g. Hyperparathyroidism was the most common histological diagnosis as severe in 13 patients (25%), or as mild in 14 (27%). Ten patients had osteomalacia (19%), adynamic bone disease was diagnosed in 5 (9.6%) and mixed renal osteodystrophy in 10 (19.2%). Low bone turnover patients showed higher bone and serum aluminum than high bone turnover patients. We observed a relative increment in high turnover bone disease in the later period (1997-2001) without changes in low turnover bone disease. These data showed a high prevalence of hyperparathyroidism and aluminum-related low turnover bone disease, with no significant changes between the two time-periods analyzed here. PMID- 12778855 TI - [Effect of strontium on bone metabolism in hemodialysis patients]. AB - The objective of this study was to assess the relationship between the bone strontium content and bone histomorphometric parameters in bone biopsies from patients with chronic renal failure undergoing hemodialysis. The study was carried out in 74 illiac crest bone biopsies from patients with renal osteodystrophy from different worldwide regions (Argentina, Portugal and Spain). They were underwent to histological and histomorphometric evaluation. The bone strontium/calcium ratio was measured by quadrupole inductively coupled plasma mass spectrometry. The samples were classified into groups according to histological criteria: hyperparathyroidism (HP), mixed (MX), osteomalacia (OM) and adynamic bone disease (ABD). Serum PTH and alkaline phosphatase before biopsy were available in most of the patients. No correlation was found between the different histomorphometric parameters and the Sr/Ca ratio. The one way ANOVA test showed statistical differences in the Sr/Ca ratio of the different histological forms (HP: 0.58 +/- 0.39; MX: 1.16 +/- 0.74; OM: 1.10 +/- 0.46; ABD: 0.91 +/- 0.40 microgram Sr/mg Ca; p < 0.003). The post-Hoc analysis showed differences between HP and MX. The biopsies having greater or equal values than 1.4 micrograms Sr/mg Ca showed higher levels of bone formation histomorphometric parameters and serum alkaline phosphatase (395 +/- 519 vs 1,022 +/- 989 UI/L, p < 0.05). Although it has been found that the biopsies with higher bone strontium had higher levels of osteoid tissue (characteristic of osteomalacia), the hypothesis of strontium-induced osteomalacia could not be demonstrated. PMID- 12778856 TI - [Role of diet in the management of osteodystrophy during progressive renal insufficiency]. AB - Secondary hyperparathyrodism (SH) is an early manifestation of chronic renal failure (CRF), which has serious complications. Moreover, treating SH is not a risk-free process. Once in its advanced state, it is extremely difficult to reverse and therefore it is critical an early intervention and prevention. An excess of phosphorus and a deficit of calcium and calcitriol are key factors in the evolution of SH. Despite the fact that plasma phosphorus levels remain normal until an extremely advanced stage of CRF, and even apparent hyperphosphatemia in mild CRF, it has been shown that restricting dietary levels of protein and phosphorus impedes the progression of SH. A decrease of protein in the diet also decreases the amount of calcium, thus the calcium levels must be supplemented in order to prevent their deficit. In addition to that slightly diminished levels of calcitriol can be observed in the early stages of CRF, thus it is logical to provide this hormone. However, administering calcitriol may induce hypercalcemia and hyperphosphatemia, which in turn risks the onset of cardiovascular calcifications and complications. Therefore, the calcitriol dosage should be small and then adjusted according to the degree of SH. Neither the PTH levels nor alterations in the phospho-calcium metabolism follow a linear increase appropriate to the decrease in renal function, therefore we propose a treatment strategy which adapts to the different degrees of renal failure. PMID- 12778857 TI - [Early treatment of secondary hyperparathyroidism in moderate renal insufficiency: low-phosphorus diet versus calcium carbonate]. AB - Calcitriol deficiency and phosphorus retention are mechanisms involved in the pathogenesis of renal hyperparathyroidism. The aim of this study was to evaluate the effect of dietary phosphorus restriction versus calcium carbonate treatment for one month on PTH and calcitriol levels in patients with mild renal failure. We studied two groups of patients: Group I: 21 patients (14M/7F); mean age 61 years old; mean glomerular filtration rate 51 ml/min. Their diet contained phosphorus 700 mg/day. Group II: 30 patients (21M/9F); mean age 58; mean glomerular rate 56 ml/min. They were divided in two subgroups: 18 patients treated with calcium carbonate 2.5 g/day and 12 patients with 5 g/day. Serum PTH, calcitriol, 25(OH)D3, calcium, phosphorus and urinary excretion of calcium and phosphorus were measured before and after a 30 day period. The low phosphorus diet (Group I) resulted in a significant decrease in PTH levels (81.3 +/- 35 vs 71 +/- 39 pg/ml, p < 0.05) and significant increase in calcitriol levels (22.4 +/ 4.4 vs 33.4 +/- 7.5 pg/ml, p < 0.05). In our study calcium carbonate treatment (Group II) had no effect on PTH and calcitriol levels. PMID- 12778858 TI - The assay of the hypocalcemic PTH fragment inhibitor with PTH provides a more accurate assessment of renal osteodystrophy compared to the intact PTH assay. AB - As the chronic kidney disease patient is being managed for PTH, calcium, phosphate, vitamin D, calcium x phosphate product and bone quality an accurate PTH measurement is essential. Over and under PTH suppressive therapies pose significant risks of mineral metabolism disturbances, osteodystrophies and soft tissue calcifications. Until recently it was thought that there was only one hormone secreted by the parathyroid gland, 1-84 PTH (or CAP). It is now known that there is another hormone secreted by the parathyroid gland (CIP) which is most likely 7-84 PTH. 7-84 PTH has been demonstrated to be an antagonist of 1-84 PTH with inverse biological activities. 7-84 PTH has been demonstrated to be hypocalcemic and able to lower bone turnover through an inhibition of osteoclast formation resulting in an overall inhibition of bone resorption. Whereas, 1-84 PTH operates through the PTH/PTHrp receptor the 7-84 PTH appears to operate through a C terminal PTH receptor. The CAP/CIP ratio decreases in the dialysis patient when calcium increases and vice versa. The 2nd generation "intact" PTH assays measure the sum of CAP plus CIP which render them ineffective at predicting bone turnover (72% predictive) and monitoring PTH suppressive treatments. By contrast the CAP/CIP ratio predicts bone turnover in the dialysis patient with a histologically determined 93% predictability. An elevated CAP/CIP ratio indicates high bone turnover and a decreased CAP/CIP ratio indicates adynamic low bone turnover. PMID- 12778859 TI - [Review of the concept of vitamin D "sufficiency and insufficiency"]. AB - There has been a poor consensus in defining normal levels of 25(OH) D. It has been traditionally recognized that 25(OH)D serum levels below 5-7 ng/ml induce osteomalacia, serum levels below 10-12 ng/ml induce secondary hyperparathyroidism and osteoporosis, and serum levels above 18-20 ng/ml are usually considered normal or adequate. Due to the results obtained in several studies, a more functional classification has recently been proposed defining serum 25(OH)D levels > 40 ng/ml or > 100 nmol/l as "desirable", serum levels between 20 and 40 ng/ml or 50 and 100 nmol/l as hypovitaminosis D, levels between 10 and 20 ng/ml or 25 and 50 mmol/l as vitamin D insufficiency and 25(OH)D levels below 10 ng/ml or 25 nmol/l as deficient. These new cut-off levels, suggest that, in the past, we had been using a wrong statistical approach for defining "normal serum 25(OH)D levels". In agreement with this new classification, in a recent study conducted in a random sample of our population, a high prevalence of low levels of 25(OH)D and secondary hyperparathyroidism was found. In our study, only in those people having "excellent" renal function, representing only 15% of the sample (serum creatinine < 1 mg/dl in men and < 0.8 in women, mean age of 68 years) hyperparathyroidism was not diagnosed despite observing 25(OH)D serum levels around 18-30 ng/ml or 45-75 nmol/l). In the remaining people (85% of the sample), who showed the expected serum creatinine increments according to their age, secondary hyperparathyroidism was avoided only if the serum 25(OH)D levels were higher than 30 ng/ml or 75 nmol/l. These remarkable findings demonstrate the importance of maintaining higher 25(OH)D levels--in addition to normal calcitriol levels--in order to avoid stimulation of the parathyroid gland. In 87 patients with a functioning renal transplantation only a 11.5% of they had levels of 25(OH)D higher than 30 ng/ml and it was correlated with PTH. These remarkable findings demonstrate the importance of maintaining higher 25(OH)D levels--in addition to normal calcitriol levels--in order to avoid stimulation of the parathyroid gland in aged people. Thus, the deficiency or even "subtle deficiency" of 25(OH)D, currently neglected in the daily management of patients with chronic renal failure, may play an important role in the maintenance of hormonal and mineral homeostasis. PMID- 12778860 TI - [Osteoporosis, estrogens, and bone metabolism. Implications for chronic renal insufficiency]. AB - The relationship between estrogens, bone metabolism and osteoporosis is well known. Chronic renal failure in women is associated with menstrual disorders, lower bone mineral density and increased risk of fractures. However, most studies on renal osteodystrophy have not taken into account the role of oestrogen deficiency, its interaction, and the possible benefits of hormone replacement therapy (HRT) in uremic women. According to these limitations and the actual evidence of benefits and risks of HRT, we conclude that: a) Osteoporosis must be evaluated as a part of renal osteodystrophy; b) HRT would be considered in women with climateric symptoms and osteoporosis, and should not be used for prevention of cardiovascular disease, and c) Clearly we need to do more studies related to osteoporosis and estrogens in CRF, but right now we have to try to optimize bone turnover in our uremic patients. PMID- 12778861 TI - [Importance of estrogen insufficiency time in the efficacy of the bone response to hormonal therapy in experimental chronic renal insufficiency]. AB - Bone disease develops relatively early in the development of CRF. The aim of this study was to evaluate the repercussion of estrogen insufficiency and the effectiveness of hormonal replacement therapy, after different periods of estrogen deprivation, on bone metabolism in an animal model with chronic renal failure and ovariectomy. A secondary purpose was to evaluate the effectiveness of bone densitometry for predicting changes in bone mass for comparison with bone histomorphometry. We used Sprague-Dawley rats with chronic renal failure and ovariectomy performed at the same time. Animals were divided into two phases according to the period of estrogen insufficiency, 4 weeks in the long estrogen insufficiency period and 1 week in the short estrogen insufficiency period. In both phases, the animals were divided into four treatment groups receiving placebo (corn oil), 17 beta-estradiol (15 micrograms/kg body weight/day), calcitriol (10 ng/kg body weight/day) or the combined treatment with estradiol and calcitriol. In both phases, a group of animals with chronic renal failure (normal ovarian function) was used as a control group. The period of treatment was 8 weeks. After this period the animals were sacrificed. This model emphasizes the importance of the period of estrogen insufficiency in the efficiency of the treatment. Four weeks of estrogen insufficiency resulted in an significant loss of trabecular bone, and less possibility of recovery. After one week of estrogen deprivation a response to the treatment was observed. The utilization of bone densitometry allowed to reproduce changes in bone mass observed afterwards by histomorphometric analysis. PMID- 12778862 TI - [Management of hyperphosphoremia]. AB - Phosphorus is involved in the pathogenesis of hyperparathyroidism in Chronic Renal Failure (CRF) and in extra-osseous calcifications. Therefore, it is directly related with the morbi-mortality of these patients. The control of phosphorus is of the utmost importance and is often extremely difficult to achieve. A quick overview of the drugs that can bind phosphorus is given in this paper. Iron salts are beginning to come through as a viable possibility in the near future; nevertheless, calcium salts continue to be the most widespread. The problem is the ease with which a positive calcium balance can be produced, as extra-osseous calcifications (and above all, vascular calcifications) can appear and morbid-mortality increases. As a result of all this, calcium-free binders such as lanthanum carbonate and sevelamer are eagerly awaited. The lanthanum is as of yet little and not very well known. Sevelamer possesses good phosphorus binding properties, as it is insoluble, does not cause intestinal degradation, does not contain traces of other elements and its use has few side effects. Long term studies and accumulated experience have demonstrated its efficacy as a binder, plus additional advantages such as an improvement in the lipid profile, with a decrease in LDL--and an increase in HDL-cholesterol. As to whether these new binders--or any other compounds that could appear-will be of any real use only time will tell. PMID- 12778863 TI - [Importance of the "adequate blood phosphorus" concept as a risk factor for hyperphosphatemia]. AB - Hyperphosphatemia is an important risk factor of secondary hyperparathyroidism and extraosseous calcifications in chronic renal failure patients. In this study our hypothesis is that physicians misconception of adequate phosphatemia is a risk factor for hyperphosphatemia. In 1999 GEMOR sent a renal osteodystrophy inquiry to different hemodialysis centers in Argentina. It included 80 dialysis centers in 17 Argentinian provinces. The enquire had 33 questions about renal osteodystrophy. Here we report the section related to phosphorous metabolism. We obtained responses from 80 dialysis centers (4,512 dialysis patients), which represents about 24% of Argentinian dialysis centers. Physicians considered phosphorous levels between 4.5 to 5.5 mg/dl in 83.5% of centers as adequate, and between 5.5 to 6.5 mg/dl in 10.1%. Five out of 77 centers reported that they had no patients with hyperphosphatemia. The percentage of hemodialysis patients that had more than 6 mg/dl in each center was 28.8 +/- 15.9%. Those centers that aimed for phosphatemia between 5.5 and 6.5 mg/dl, had a higher percentage of patients with phosphatemia above 6 mg/dl than those aiming for between 4.5 and 5.5 mg/dl (42.8 +/- 16.7 vs 27.1 +/- 15.2% respectively, p = 0.007), and had higher mean of phosphatemia (6.4 +/- 0.7 vs 5.3 +/- 0.7 mg/dl respectively, p = 0.0001), than the last group. In conclusion, a higher mean phosphate level was obtained in hemodialysis centers where physicians considered higher pre-dialysis target levels. Some centers had no patients with hyperphosphatemia (neglect or good control?). PMID- 12778864 TI - [Estimation of bone mass of hemodialysis patients by digital radiologic radiogrammetry (DXR)]. AB - Bone mass (BMD) may be evaluated by digital X-ray radiogrammetry (DXR) which it is estimated from multiple cortical measures carried out on five regions of interest over a simple hand-forearm X-ray film. We included 168 HD patients (108 men and 60 women; 63 +/- 14 and 66 +/- 12 years old respectively--p < 0.05-) coming from seven HD units in Asturias. We performed a hand-forearm X-ray, an epidemiologic questionnaire and we revised their medical and analytical records. As the normative data we used the measurements of a random sample of 247 subjects from our own population. The prevalence of osteoporosis was 7% and 40% in men and women respectively. BMD was correlated with weight (r = 0.346), time on HD (r = 0.188), time on treatment (r = -0.235). The porosity was correlated with PTH levels. PMID- 12778865 TI - [Prevalence of vertebral fractures and aortic calcifications in hemodialysis patients: comparison with a population of the same age and sex]. AB - Dialysis patients have bone metabolic disorders and a higher prevalence of fractures, principally peripheral fractures. However, there are few studies focusing on the prevalence of vertebral fractures. Moreover, aortic calcifications are very common and are an independent predictive factor of vascular morbidity and mortality. The objective of this study was to assess the prevalence of vertebral fractures and vascular calcifications in haemodialysis (HD) patients (n = 99), in comparison with a random sample of general population of similar age and from the same geographical area (n = 624) and study their relationship with clinical, biochemical and therapeutical data. The prevalence of vertebral fractures in HD patients and general population was 19.1% and 24.1% respectively (non-significant statistical differences). In both, sexes, the presence of vertebral fractures was positively associated with age, mean maximum Ca, mean maximum CaxP. In women, time in HD was positively associated as well. On the other hand, the prevalence of aortic calcifications was much higher in HD patients (77.9% vs 37.5%, p < 0.001). HD was a risk factor for aortic calcification in women [OR = 7.7 (IC 95% = 2.6-22.9)] as in men [OR = 5 (IC 95% = 1.9-12.9)]. Severe vascular calcifications were more frequent in HD patients, it reached 57.4% compared with 17% of general population (p < 0.001). Both, in women (64.5% vs 13.3% p < 0.001) and in men (51.4% vs 20.9%), respectively (p < 0.001). In conclusion, the prevalence of vertebral fractures was similar in HD patients and in general population. Nevertheless, frequency and severity of aortic calcifications was higher in HD patients. PMID- 12778866 TI - [Vascular and tissue calcifications of hemodialysis patients]. AB - The present paper aims to assess radiographic vascular and soft parts calcifications occurrence and its correlation with biochemical profiles. The study was performed in 47 patients (ten diabetic patients), 49 years old, who had been on dialysis for a period of 51 months. Vascular calcifications (VCs) were classified as proximal, distal and soft tissues. In addition, Ca, P, CaxP values in the six months prior to the recruitment period, PTH, FAL and calcium carbonate, calcium acetate, and vitamin D3 intake were determined. A higher frequency of VCs was observed in diabetics, yielding a significant association with proximal 60% (p = 0.05) and almost significant with distal calcifications 70% (p = 0.07). Likewise, a lower CaxP was noted for diabetic VCs in comparison to that seen in non-diabetic VCs (p < 0.05). Proximal and distal VCs in the non diabetics population were 25% and 20%, respectively; and tissue calcifications were 24%. Age was correlated with proximal and distal VCs (p < 0.01). A higher CaxP was observed in patients with VCs and it yielded an even higher value for tissue calcifications. Lastly, calcium acetate and overall calcium intake was higher in patients with tissue calcifications (p = 0.05). VCs were more frequent in diabetics and they also showed a lower CaxP. VCs in non-diabetics were correlated with CaxP values, whereas tissue calcifications were associated with calcium intake. Therefore, the management of renal osteodystrophy should be changed in order to prevent calcifications as well as to decrease morbidity in hemodialysis patients. PMID- 12778867 TI - [Differential effect of vitamin D analogues on the proliferation of vascular smooth muscle cells]. AB - Atherosclerosis is the principal cause of myocardial infarction, stroke, and peripheral vascular disease, accounting for nearly half of all mortality in developed countries. The excessive growth of vascular smooth muscle cells is an important component in the development of atherosclerotic lesion. The direct effect of calcitriol and vitamin D analogs on the VSMCs proliferation is not clear. In this study we have analysed if calcitriol, Paricalcitol (19-nor-1,25 dihydroxy-vitamin D2) and EB1089 (experimental analog used as anticancerous) modify proliferation and the expression of vitamin D receptor (VDR) gene that is regulated at the transcriptional level by itself in the VSMCs. VSMCs proliferation was analysed by BrdU incorporation and VDR gene expression using RT PCR. VSMCs proliferation was stimulated when calcitriol was added to the culture. VSMCs proliferation was significantly lower with analogs at the same dose. With regard to the functional study, the expression of VDR gene was upregulated by calcitriol at a concentration of 100 nM. There were no changes in this expression with the analogs. In conclusion, calcitriol, do not modify VSMCs proliferation. Therefore, Paricalcitol could have a minor proliferating effect on the wall of vessels that vitamin D. PMID- 12778868 TI - [Bone metabolism alterations after kidney transplantation]. AB - Early after renal transplantation (RT) a rapid decrease in bone mineral density at the lumbar spine, femoral neck, and femoral shaft has been documented. In addition, an appreciable proportion of patients still remain losing bone late after RT. As a consequence, RT patients are at a high risk of bone fractures as compared to general population. Most fractures involve appendicular skeleton, particularly the feet and ankles, and the diabetic patient is at increased risk of fractures. Thus, early institution of preventive measures and treatment of established osteoporosis are central. The major cause of post-transplantation bone loss is corticosteroid treatment, and this should be used at the lower dose compatible with graft survival. Preexisting hyperparathyroidism also affects the early cancellous bone loss at the spine, and post-transplantation bone loss reflects variable individual susceptibility, resembling the polygenic determination of bone mineral density in general. Clinical trials have demonstrated that bisphosphonates or vitamin D plus calcium supplementation, prevent post-transplantation bone loss during the first 6-12 months. However, their role in preventing bone fractures has not been proven. Finally, recommendations for management, prevention and treatment, are summarized. PMID- 12778869 TI - Osteoporosis after renal transplantation. AB - This study was performed to determine risk factors associated with osteoporosis that develops after renal transplantation. Sixty-five kidney graft recipients were included in this study. They were divided into four groups according to the time since transplantation: Group 1 (< 1 year; n = 26), group 2 (1-3 years; n = 16), group 3 (3-5 years; n = 12) and group 4 (> 5 years; n = 11). These groups were matched according to probable risk factors for osteoporosis, findings of serum biochemistry, biochemical markers of bone turnover and measurements of bone mineral density. One way ANOVA test and Kruskal-Wallis test were used for statistical analysis. Osteoporosis was found in 22 recipients (33.8%). There were significant differences in recipient age, cumulative steroid dose, and episodes of acute rejection between the four groups. Increasing age, cumulative steroid dose and episodes of acute rejection were found to be risk factors for osteoporosis in our study. PMID- 12778870 TI - Bone loss in pediatric renal transplant recipients. AB - The factors that affect bone mineral density (BMD) and the long term progress of BMD after transplantation in children is still unknown. Therefore we performed a cross-sectional study to determine BMD in 83 recipients who received living renal allotransplants in Mansoura Urology & Nephrology Center between 1981 and 2001 (mean age at transplantation 13.2 +/- 3.1 years) by dual energy x-ray absorptiometry at various time intervals up to 16 years after transplantation (mean duration after transplantation was 48 +/- 34 months, range 6-192 months). The Z-score for lumbar spine was -2.28 +/- 2.06 and -1.44 +/- 1.44 for the total body. Osteopenia/osteoporosis were present in about two thirds of our kidney transplant recipients. The significant predictors for osteopenia/osteoporosis by univariate analysis were cyclosporine based immunosuppression, the cumulative steroid dose/m2 surface area, graft dysfunction and the urinary deoxypyridinoline. Using logistic regression analysis the cumulative steroid dose/m2 surface area and the urinary deoxypyridinoline were the major significant predictors for bone loss. PMID- 12778871 TI - [New reflections on reno-vascular medicine]. PMID- 12778872 TI - [ANCA vasculitis]. PMID- 12778873 TI - [Transplants in Italy: times are changing]. PMID- 12778874 TI - [Emerging features in acute renal failure]. PMID- 12778875 TI - [Multicenter study of darbepoetin alfa in the treatment of anemia secondary to chronic renal insufficiency on dialysis]. AB - This Spanish single-arm, multicenter, prospective clinical trial assessed the maintenance of hemoglobin concentrations (Hb) between 10-13 g/dL with unit doses of darbepoetin alfa and the safety of the treatment in dialysis patients. Eight hundred twenty-six patients with chronic renal failure (CRF) (94% receiving haemodialysis and 6% receiving peritoneal dialysis) previously maintained on stable recombinant human erythropoietin (r-HuEPO) therapy with stable hemoglobin (Hb) concentrations (mean Hb concentration = 11.7 g/dL) were switched to darbepoetin alfa at a reduced dosing frequency for 24 weeks (a 20-week titration phase plus a 4-week treatment evaluation phase). Subjects receiving r-HuEPO two or three times weekly were switched to darbepoetin alfa once weekly, and those. who were receiving r-HuEPO once weekly were switched to darbepoetin alfa once every two weeks. The initial dose of darbepoetin alfa was determined from the r HuEPO dose at inclusion into the study using a formula equating the peptide mass of the two molecules and rounding to the nearest available prefilled syringe dose. Overall, 86.8% of patients completed the 24-weeks of study. Changing the treatment from r-HuEPO to darbepoetin alfa and increasing the dose interval did not result in any clinically significant change in the Hb concentration. From base-line to the evaluation phase, the mean Hb fell 0.09 (95% CI, -0.2; -0.0) g/dl, with an increase of 0.19 (95% CI, 0.0;0.3) g/dL i.v. and a decrease of 0.22 (95% CI, -0.3; -0.1) g/dL s.c.). This maintenance of the mean Hb concentration was accompanied by a mean 9.8% reduction of the darbepoetin alfa dose (19.7% (95% CI, -24.9; -14.2) i.v. and 4.7% (95% CI, -8.5; -0.7) s.c. Treatment with darbepoetin alfa was well tolerated and no unexpected adverse events were reported. In conclusion, the replacement of previous r-HuEPO treatment by darbepoetin alfa in the therapy of anemia secondary to chronic renal failure in diaiyzed patients was effective, well tolerated, and decreased the frequency of dose administration compared with the previous r-HuEPO treatment. Darbepoetin alfa administered once weekly or once every two weeks maintained the baseline Hb levels whilst allowing dose reduction, which was higher in patients receiving i.v. darbepoetin alfa. PMID- 12778876 TI - [Immunoallergic interstitial nephritis vs. cholesterol atheroembolism. Differentiating characteristics]. AB - The commonest clinical presentation of both immunoalergic interstitial nephritis (IIN) and atheroembolic renal disease (ATD) is an acute renal failure accompanied by skin lesions and eosinophilia. As a consequence, differential diagnosis between both entities is often very difficult. We have performed a comparative retrospective study of those patients diagnosed as having IIN or ATD in our Hospital in the period 1980-2000. A total of 42 patients have been diagnosed of IIN and 16 of ATD. Demographic data, as well as clinical and laboratory parameters and outcomes of every studied patient were analysed. We found a significantly higher prevalence of male sex (100% vs 57%, p < 0.01), previous history of hypertension (100% vs 55%, p < 0.01), chronic renal insufficiency (56% vs 17%, p < 0.01), ischemic heart disease (56% vs 14%, p < 0.001), peripheral ischemic disease, endovascular procedures (87% vs 7%, p < 0.001) and anticoagulant treatments (25% vs 5%, p < 0.001) among patients with ATD as compared with IIN, respectively. On the contrary, previous infections (45% vs 12%, p < 0.01) and exposure to new drugs (100% vs 40%, p < 0.001) were significantly more frequent among IIN patients in compare with ATD. ATD patients showed skin lesions consisting of livedo reticularis and digital infarcts (63% vs 31%, p < 0.05) accompanied by blood pressure increase (100% vs 24%, p < 0.001), whereas IIN patients showed fever (41% vs 19%, p < 0.05) and cutaneous rash as significant clinical manifestations, respectively. The number of ATD patients with proteinuria > 1 g/24 h was significantly higher, but no differences between both groups in the prevalence of urinary sediment abnormalities were observed. The prevalence of absolute eosinophilia was high in both groups (88% among ATD patients, 64% among IIN patients; pNS). Prognosis of both entities was clearly different: Almost all patients with ATD died (69%) or evolved to end-stage renal failure, whereas most patients with IIN showed a recovery of renal function after withdrawal of responsible drugs and steroid treatment. In summary, the analysis of clinical and laboratory data allows an initial differential diagnosis in patients suspected as having IIN or ATD. PMID- 12778877 TI - [Survival of myeloma patients treated with dialysis]. AB - BACKGROUND: Renal failure is a common complication of myeloma. Renal replacement therapy in these patients is controversial due to poor survival outcomes and low tolerance to treatment. We reviewed our experience on patients with myeloma undergoing dialysis therapy at one centre. PATIENTS AND METHODS: Between 1980 and 2000, 28 patients (21 men and 7 women) with myeloma were admitted to chronic dialysis programme and the following variables were analysed: sex, age when starting dialysis, lapse of time between diagnosis of myeloma and admission to dialysis (TD), disease stage, comorbity, mode of presentation, calcium, creatinine at diagnostic, albumin and Hb at the beginning of dialysis, and cause of death. We studied survival among these patients (Kaplan-Meier), identified predictors of survival outcome (Cox's regression) and compared survival between the two decades studied. RESULTS: Mean age was 65 years, median TD was 0.4 months, and modes of presentation were: end-stage renal failure (18 patients), acute renal failure (8), amyloldosis (2). Eleven patients (39%) had myeloma IgG, four (14%) IgA and thirteen (46%) had light chains. Kappa light chain was the most frequent one. In 75% of patients myeloma was at IIIb stage. Cause of death were: Cardiovascular disease (5 patients), infections (4), suspension of treatment (4), tumours (4), and others causes (2). Median survival for all patients was 16.8 months (range 0.4-78) and 25% survived over 39 months. Hb level was the only significant predictor in the multivariant analysis (p = 0.02). In the 80's median survival was 6.17 months versus 17 months in the 90's but this difference was not significant with long-rank test. CONCLUSION: Although survival of patients with myeloma treated with dialysis is still short, 25 percent survive over 3 years, being Hb level the only predictive factor. Moreover, we observed an improvement of survival in recent years. PMID- 12778878 TI - [Effect of arterial pressure and age on renal function, The "Care for the Kidney" study]. AB - BACKGROUND: The kidney is one of the principal target organs of hypertension. The mechanism by which hypertension damages the kidney and the relative contribution of high blood pressure to the progression of renal failure remains incompletely defined. The clinical quantification of renal function is usually thought to require determination of plasma clearance of endogenous creatinine (ClCr), an impractical test for epidemiological evaluation. For this reason several formulae have been proposed to estimate the ClCr from the serum creatinine concentration, such as sex, age, and body weight. The most often used formula is the one proposed by Crockroft and Gault in 1976. OBJECTIVE: To determine the prevalence of loss of renal function and the influence of hypertension and age on this loss in hypertensive and normotensive patients. PATIENTS AND METHOD: An observational, cross-sectional comparative study was carried out. 3,420 patients (1,171 normotensive and 2,249 hypertensive) were studied. Average age was 70 years (19.4% of patients < 65 years; 23.3% > 75 years). 98.1 of hypertensive patients were treated with drugs, 26.7% of them with more than one drug. 27.9% of treated patients achieved blood pressure control (< 140-90 mm Hg). The level of renal function was estimated by means of ClCr using the Crockroft and Gault formula. RESULTS: 3.9% of normotensive and 12.2% of hypertensive patients had serum creatinine values > 1.3 mg/dl, but 28.6% of normotensive and 40.4% of hypertensive had ClCr < 60 ml/min. The patients over 75 years had ClCr mean values < 60 ml/min. There was correlation between ClCr and age, but there was no correlation between ClCr and blood pressure values. CONCLUSIONS: The estimated ClCr decrease with age and there existed correlation between both variables. The ClCr is lower in hypertensive than in normotensive patients. Hypertension and age influence loss of renal function, but age is a determining factor. The estimated ClCr was a more sensible method of evaluating the loss of renal function than the serum creatinine values. The use of Crockroft and Gault formula can be useful for making diagnostic and therapeutic decisions in primary care. PMID- 12778879 TI - [Self-esteem in Mexican pediatric patients on peritoneal dialysis and kidney transplantation]. AB - It has been reported in literature that self-esteem increases in patients who pass from continuous ambulatory peritoneal dialysis to renal transplantation. However, our clinical observations in the Department of Nephrology of Hospital Infantil de Mexico Federico Gomez indicate otherwise. Therefore, we carried out this research on 60 patients, children and adolescents, male and female between the ages from 8 to 15 years old, and we applied them the Coopersmith self-esteem questionnaire, which has been validated in mexican population. These findings indicate that the social environment continuous ambulatory peritoneal dialysis patients feel more adapted than patients in renal transplant (p = 0.05). However it was also observed that these patients tried to show a better image on their situation through compensatory mechanisms (defense p = 0.03). It was concluded that it would be necessary to follow the adaptation of the transplanted patients, in order to help them achieve complete rehabilitation. PMID- 12778880 TI - [Atherosclerotic renovascular hypertension: clinical findings and results of treatment over 15 years]. AB - The aim of this study was to present our clinical experience and results of different treatments in 83 atherosclerotic renovascular hypertensive patients treated in the last 15 years in the Instituto de nefrologia in Havana. Regardless of the type of treatment the patients were divided in two groups. Group I: 52 (62.3%) cases with standard oral hypotensive drugs alone and control of other cardiovascular risk factors (mean age 53 years old, sex m/f 50/50%, race white/no white 75/25%, mean known hypertension follow-up 10.2 +/- 10 years, mean SBP 208 +/- 30 mmHg, mean DBP 123 +/- 17 mmHg, mean serum creatinina 1.62 mg/dl and increase peripheral plasma renin value in 61.6% of patients) and group II: 31 (37.7%) cases treated with revascularizacion procedures (PTA or surgery) or nephrectomy in selected patients (mean age 50 years old, sex m/f 68/32%, race white/no-white 16/84%, mean known hypertension follow-up 8.5 +/- 8.6 years, mean SBP 214 +/- 32 mmHg, mean DBP 1.31 +/- 16 mmHg, mean serum creatinina 1.85 mg/dl and increase peripheral plasma renin value 78.3% of patients). As end point for treatment results we selected: 1) hypertension cure or control, 2) evolution of the serum creatinine value and 3) kidney and patients survival. RESULTS: In those cases with a follow up for more than one year, in 82.9% the blood pressure was cure (21.4%) or controlled (61.4%). The proportion of failed was superior in group I (20.9%) than in group II (11.1%). All 18 cases treated by PTA with a follow up period longer than a year, blood pressure cure in 10 (55.6%), ameliorate in 5 (27.8%) and in 3 (16.6%) was unchanged (one patient lost of follow up). Nine patients were treated by surgery (3 revascularization and 6 nephrectomy), 5 (55.5%) cases cured and 4 (44.5%) ameliorate his blood pressure. Patients in group II maintain normal renal function in more cases than in group I (48.4% vs 30.8%). Both group had similar percentage of normal-normal + pathology normal renal function (G I: 65.4% vs G II: 77.4%) p = 0.29. When chronic renal function was present at the base line study none of the revascularization procedure were superior. Patient and Kidney actuarial survivals rate do not showed superiority for any treatment procedure after 10 years of follow up. CONCLUSIONS: In atherosclerosis renovascular hypertension patients treated with intervention procedure had better BP control than those treated by hypotensive drugs. Not significant different between intervention procedures and drugs treatment in renal function preservation or in patient and kidney actuarial survival rate were found in these patients. PMID- 12778881 TI - [Evaluation of the oblique supine position for percutaneous renal biopsy]. PMID- 12778882 TI - [Presence of hepatitis C virus in renal tissue in membranoproliferative glomerulonephritis and cryoglobulinemia]. AB - Although hepatitis C virus infection has been documented in several extrahepatic diseases, the deposition of HCV RNA in glomerular structures has proved to be difficult to demonstrate. We report a patient with membranoproliferative glomerulonephritis, type III circulating cryoglobulins and hepatitis C virus infection with detection of HCV RNA in serum, cryoprecipitate and renal tissue using specific RT-PCR technique. These data confirm that HCV could have a direct role in renal damage. PMID- 12778883 TI - [Non-invasive monitoring of a humeral arteriovenous fistula with hemodynamic repercussions]. AB - A 70-year-old woman, who undergoing hemodialysis due to chronic pyelonephritis, is reported. She suffered from dyspnea due to pulmonary artery hypertension secondary to volume overload as a complication of high-flow brachial AVF. The combined estimation of vascular access blood flow rate (QA) and systolic pulmonary artery pressure by noninvasive methods (ultrafiltration and Doppler echocardiography, respectively) allowed us the diagnosis, make a surgical indication and post-surgical follow-up of AVF with hemodynamic repercussion. PMID- 12778884 TI - [Familial focal and segmentary hyalinosis]. AB - Focal segmental glomerulosclerosis represents a finding in several renal disorders, characterized by proteinuria and sometimes by arterial hypertension and progressive decline in renal function. There are primary (idiopathic and familial) and secundary forms. In the last 20 years several familial cases has been reported, with a great genetic heterogeneity (dominant and recessive forms) and with multiple associations with particular MHC class-I and class-II gene loci, being Al, DR3 o DR7 the most frequently reported. We described three members of same family with focal segmental hyalinosis that shared the HLA haplotype A31 B61 DR13. This association has not been described previously. We highlight that genetic and acquired factors (obesity, hypertension...) could have importance in the development of progressive renal failure in these patients. PMID- 12778885 TI - [A hematocrit greater than 36 lessens mortality and hospitalization in hemodialysis]. PMID- 12778886 TI - [High prevalence of sebaceous hyperplasias in renal transplants]. PMID- 12778887 TI - [Changes in iron metabolism and erythropoietin requirements after the switch from ferric gluconate to iron saccharose. Is it worth the increased expense?]. PMID- 12778888 TI - [Functional glycoconjugates involved in cellular interaction]. AB - We first examined the involvement of the complex sphingolipids in cell-substratum adhesion using GM-95, a mutant cell line deficient in glycosphingolipids (GSLs) due to the lack of ceramide glucosyltransferase activity. We determined the adhesion of the mutant cells and stable transfectants expressing GSLs, which were established by transfection of GlcT-1 cDNA into GM-95 cells under neutral sphingomyelinase (sm) treatment. We confirmed that complex sphingolipids play critical roles in cell-substratum adhesion, and the presence of either GSLs or SM is sufficient for the adhesion. We also investigated intracellular signaling (glycosignaling) mediated by endogenous GM1a involved in the neuronal differentiation of PC12 cells using the cholera toxin B subunit (CTB) that specifically binds to ganglioside GM1a. Treatment with CTB induced neuron-like differentiation of PC12 cells. Biochemical analyses demonstrated that the tyrosine phosphorylation induced by CTB was responsible for neuron-like differentiation of PC12 cells and that the MEK-ERK cascade is a part of the biological signals mediated by endogenous ganglioside GM1a on PC12 cells. We further demonstrated that glycosignaling is mediated through a high-affinity ligand, PSGL-1, for P-selection on neutrophils. In this case, engagement of PSGL 1 on the cell surface strongly induced tyrosine phosphorylation of several cellular proteins including ERKs and activated a canonical MAP kinase pathway. Tyrosine phosphorylation induced by engagement of PSGL-1 is responsible for the secretion of interleukin-8 from neutrophils, suggesting that PSGL-1-mediated glycosignals are involved in the progression of the inflammatory response. In this review, we mainly discuss the biological and pathological significance of glycoconjugates in relation to the above issues. PMID- 12778889 TI - [Role of metabosensitive afferent fibers in neuromuscular adaptive mechanisms]. AB - Role of metabosensitive afferent fibers in neuromuscular adaptive mechanisms. Adaptation to exercise is provided by central neuron activity adjustments which are regulated partly by activation of group I and II (mechanosensitive) and group III and IV (metabosentitive) afferent fibers. These last two groups are activated by exercise-induced changes in muscle metabolism. The role played by these afferents seems to be crucial to exercise and fatigue tolerance adaptive mechanisms. Nevertheless, many questions remain unresolved. The aim of this review is to focus on the involvement of metabosensitivity in sensorimotor loops and neuromuscular adaptive mechanisms. The existence of an adaptive cardiovascular and respiratory reflex to exercise originating from metabosensitive afferent fiber activation is well established. Furthermore, the mechanism of skeletal muscle protection against fatigue could be due to modulation of central motor command at the spinal and supraspinal levels via these afferent fibers. PMID- 12778890 TI - Validation of three alternative methods to measure total energy expenditure against the doubly labeled water method for older Japanese men. AB - In a previous study using young Japanese men as subjects, Ebine et al. found that accelerometer (AC) represents a promising technique for measuring free-living total energy expenditure (TEE) when compared to activity records (AR) and heart rate monitoring (HR). Thus, the present study was designed to validate the use of an AC and to determine whether or not the previous findings regarding the three alternative field methods (AC, AR, and HR) could be extended to older Japanese men (n = 24; mean +/- SD age 48 +/- 10 y, body mass index 23.1 +/- 2.7 kg/m2 and body fat 18.7 +/- 4.8%). TEE values obtained over a 3 d period by AR, HR, and AC (3dAC), and AC over a 14 d period (14dAC) were simultaneously validated against TEE measured by the doubly labeled water (DLW) method applied within a 14 d period. TEE values obtained by AR, HR, 3dAC, and 14dAC ranged from 1,750 to 3,447 kcal/d, 1,691 to 5,286 kcal/d, 1,716 to 2,765 kcal/d, and 1,700 to 2,855 kcal/d, respectively. Expenditures obtained by HR were similar to those obtained using the DLW method, with a mean difference of 57 +/- 603 kcal/d (2%), but those obtained using AR, 3dAC, and 14dAC differed substantially from the DLW method, with mean differences of -335 +/- 289 kcal/d (12%), -542 +/- 249kcal/d (-19%), and -566 +/- 223kcal/d (-20%), respectively. AR, HR, 3dAC, and 14dAC were significantly correlated with the DLW method, with r values of 0.76 (p < 0.0001), 0.67 (p < 0.001), 0.78 (p < 0.0001), and 0.83 (p < 0.0001), respectively. Intra individual variation indicated by the coefficient of variation (CV) was significantly higher for HR (15 +/- 11%, p < 0.001) than for AR (7 +/- 4%), 3dAC (7 +/- 5%), and 14dAC (8 +/- 31%). The same findings were obtained using Bland and Altman plots at the population level. Interestingly, 3dAC and 14dAC were significantly correlated with r = 0.97 (p < 0.0001), with a lower mean difference of 24 kcal/d. These results suggest that, same as the previous study, AC is superior to HR in estimating TEE, and seems to be satisfactory for estimation at both group and individual levels, particularly for large-scale studies of older individuals when compared to the DLW method. However, some modifications of the AC method may be needed to compensate for the underestimation of TEE. PMID- 12778892 TI - Current awareness in prenatal diagnosis. PMID- 12778891 TI - Seasonal changes in immune response and parasite impact on hosts. AB - Seasonal changes in the impact of parasites on hosts should result in seasonal changes in immune function. Since both ectoparasites and endoparasites time their reproduction to that of their hosts, we can predict that hosts have been selected to show an annual peak in their ability to raise an immune response during the reproductive season. We found large seasonal changes in immune function between the breeding and the nonbreeding season for a sample of temperate bird species. These changes amounted to a decrease in spleen mass from the breeding to the nonbreeding season by on average 18% across 71 species and a seasonal decrease in T-cell-mediated immunity by on average 33% across 13 species. These seasonal changes in immune function differed significantly among species. The condition dependence of immune function also differed between the breeding and the nonbreeding season, with individuals in prime condition particularly having greater immune responses during breeding. Analyses of ecological factors associated with interspecific differences in seasonal change of immune function revealed that hole-nesting species had a larger increase in immune function during the breeding season than did open nesters. Since hole nesters suffer greater reduction in breeding success because of virulent parasites than do open nesters, this seasonal change in immune function is suggested to have arisen as a response to the increased virulence of parasites attacking hole-nesting birds. PMID- 12778893 TI - AARC clinical practice guideline. Bland aerosol administration--2003 revision & update. PMID- 12778894 TI - AARC clinical practice guideline. Capnography/capnometry during mechanical ventilation--2003 revision & update. PMID- 12778895 TI - AARC clinical practice guideline. Intermittent positive pressure breathing--2003 revision & update. PMID- 12778896 TI - [One-sided spinal anesthesia--a beautiful, but future concept? AINS 2002; 37:649 650]. PMID- 12778897 TI - Lives of the cell. AB - What is the relation between things and theories, the material world and its scientific representations? This is a staple philosophical problem that rarely counts as historically legitimate or fruitful. In the following dialogue, the interlocutors do not argue for or against realism. Instead, they explore changing relations between theories and things, between contested objects of knowledge (like the cell) and less contested, more everyday things (like frog eggs scooped from a pond). Widely seen as the life sciences' first general theory, the cell theory underwent dramatic changes during the nineteenth century. The dialogue established that each successive version of the cell theory was formulated - each identity of the object cell was formed - around a different material: cork, cartilage, eggs in cleavage, muscle. Such things thus serve as exemplary materials, in ways not described by standard concepts like induction, theory testing, theory-laden observation, and construction. Still, how can theories and perspective possibly be honed on things if these are apprehended differently by different observers according to their interests, training, culture, or indeed theories? The second part of the dialogue addresses this problem, partly through the verbal and visual schemata that were used by nineteenth-century microscopists and that are comparable to schemata in the visual arts. The third part of the dialogue considers the exemplary materials as a historical sequence, itself needing explanation. Theoretical change devolved partly from wider histories and geographies of the prevalence, availability, or scientific and cultural status of materials such as plants, animals, and muscle. PMID- 12778898 TI - More fibre: the negotiation of microscopic facts in Victorian Britain. AB - During the 1840 and 1850s the British embryologist and histologist Martin Barry (1802-1855) propounded a bold and original thesis about the microscopic structure of animal and vegetable tissue. He maintained that minute double spirals were virtually ubiquitous in the makeup of a wide range of structures. This paper considers how a claim of this kind was consonant with a romantic image of scientific creativity with which Barry identified. It describes his partially successful strategies to convince contemporaries of the veracity of his claims. Major figures in the field, such as Richard Owen and Jan Evangelista Purkyne, affirmed that Barry's spirals were real objects in nature. Others, notably William Sharpey, became convinced that the spirals were mere artifacts and that Barry was deeply flawed as a scientific investigator. The ultimate rejection of his hypothesis had much to do with the moral repugnance that Barry's attempts to gain credit for a major discovery evoked among influential medical scientists. This negative assessment of Barry as an investigator reveals the lineaments of an alternative ethic of scientific practice. PMID- 12778899 TI - Linkage: from particulate to interactive genetics. AB - Genetics was established on a strict particulate conception of heredity. Genetic linkage, the deviation from independent segregation of Mendelian factors, was conceived as a function of the material allocation of the factors to the chromosomes, rather than to the multiple effects (pleiotropy) of discrete factors. Although linkage maps were abstractions they provided strong support for the chromosomal theory of inheritance. Direct Cytogenetic evidence was scarce until X-ray induced major chromosomal rearrangements allowed direct correlation of genetic and cytological rearrangements. Only with the discovery of the polytenic giant chromosomes in Drosophila larvae in the 1930s were the virtual maps backed up by physical maps of the genetic loci. Genetic linkage became a pivotal experimental tool for the examination of the integration of genetic functions in development and in evolution. Genetic mapping has remained a hallmark of genetic analysis. The location of genes in DNA is a modern extension of the notion of genetic linkage. PMID- 12778900 TI - The article of Shertzer et al. (2002) has several short comings. PMID- 12778902 TI - Stem cell transplant regenerates neuronal cells in mice. PMID- 12778901 TI - Non-steroidal anti-inflammatory drug users possibly have a decreased risk of Alzheimer's disease. PMID- 12778903 TI - Clinical impact and variable distribution of sleep disorders among the epilepsies. PMID- 12778904 TI - Researcher's study whether Parkinson's disease impacts body mass index. PMID- 12778905 TI - Unraveling Reaven's syndrome X: serum insulin-like growth factor-I and cardiovascular disease. PMID- 12778906 TI - Unraveling Reaven's syndrome X: serum insulin-like growth factor-I and cardiovascular disease. PMID- 12778907 TI - Polyboroides radiatus predation attempts on Propithecus verreauxi. PMID- 12778908 TI - Vaginal fatty acids produced by chimpanzees during menstrual cycles. PMID- 12778909 TI - Association between Galago thomasi and Pan troglodytes schweinfurthii in the Kibale National Park, Uganda. PMID- 12778910 TI - Observation of a sudden resident-male replacement in a unimale bisexual troop of Hanuman langurs, Semnopithecus entellus, around Jodhpur (India). PMID- 12778911 TI - You can't get there from here: a case comment on Arndt v. Smith. PMID- 12778912 TI - Physician-assisted suicide: the Hippocratic dilemma. PMID- 12778913 TI - Altitudinal ranging of Rhinopithecus bieti at Jinsichang, Lijiang, China. PMID- 12778914 TI - Mother may I ... live? Parental refusal of life-sustaining medical treatment for children based on religious objections. PMID- 12778916 TI - Harming future persons: obligations to the children of reproductive technology. PMID- 12778915 TI - Silverback male presence and group stability in gorillas (Gorilla gorilla gorilla). PMID- 12778917 TI - Protecting children against unnecessary institutionalization. PMID- 12778918 TI - Reactions of chimpanzees and gorillas to human observers in a non-protected area in south-eastern Cameroon. PMID- 12778920 TI - Observed infanticide in a seasonal breeding prosimian (Propithecus verreauxi verreauxi) in Kirindy Forest, Madagascar. PMID- 12778919 TI - Patient advocates or patient adversaries? Using fiduciary law to compel disclosure of managed care financial incentives. PMID- 12778921 TI - Pegram v. Herdrich: HMOs aren't perfect, but ... PMID- 12778922 TI - The rebirth of informed consent: a cultural analysis of the Informed Consent Doctrine after Schreiber v. Physicians Insurance Co. of Wisconsin. PMID- 12778923 TI - A new subspecies of golden langur (Trachypithecus geei) from Bhutan. PMID- 12778924 TI - Stopping the baby-trade: affirming the value of human life through the invalidation of surrogacy contracts: a blueprint for New Mexico. PMID- 12778925 TI - Moore 10 years later--still trying to fill the gap: creating a personal property right in genetic material. PMID- 12778926 TI - White-faced Capuchins cooperate to rescue a groupmate from a boa constrictor. PMID- 12778927 TI - Redefining family: should lesbians have access to assisted reproduction? AB - The recent Federal Court decision in McBain v. Victoria, which rendered inoperative a Victorian law that restricted assisted reproductive technology to married or heterosexual de facto couples, has raised the issue of whether lesbians should have access to such technology. This article provides an overview of State laws currently regulating lesbian access to assisted reproduction in Australia. It then explores the growing body of empirical research indicating that the welfare of children raised in lesbian households does not differ in any significant respect from the welfare of children raised in comparable circumstances by heterosexual parents. This research undermines the view that children suffer social stigma or experience hardship caused by the lack of a 'father figure.' The 'welfare of child' rhetoric has in fact been used to mask marginalisation of 'alternative' family forms, and the reluctance to extend assisted reproductive technology to lesbians is underpinned by a deep-rooted fear of undermining the traditional heterosexual nuclear family. PMID- 12778928 TI - Termination of pregnancy for reason of foetal disability: are there grounds for a special exception in law? PMID- 12778929 TI - The road to elucidating the mechanism of manganese-bilirubin-induced cholestasis. PMID- 12778930 TI - Genetic secrets and the family. PMID- 12778931 TI - Genetic secrets and the family: a response to Bell and Bennett. PMID- 12778932 TI - Beaten before they are born: immigrants, their children, and a right to prenatal care. PMID- 12778933 TI - The Hope Clinic v. James E. Ryan. PMID- 12778934 TI - Mentoring the mentors: the Yoda factor in promoting scientific integrity. PMID- 12778935 TI - The pressing need for postdoctoral research ethics education. PMID- 12778936 TI - A rose by any other name is still a rose. PMID- 12778937 TI - One field, many disciplines, one goal. PMID- 12778938 TI - A lawyer's perspective on graduate studies in bioethics. PMID- 12778939 TI - Medical ethics and the moral practice of medicine. PMID- 12778940 TI - In focus. Core faculty and their publications at bioethics centers in the United States. PMID- 12778941 TI - In focus. Has patient autonomy gone to far? Geneticists' views in 36 nations. AB - We surveyed genetics professionals, patients, and the public about rights to information, to requested services, and to parenthood, posing difficult cases found in practice. In all, 2906 genetics professionals (63%), 499 primary care physicians (59%), 476 North American genetics patients (67%), 394 French patients (51%), 593 German patients (65%), and 988 members of the American public (99%) returned anonymous questionnaires. Results suggest a trend toward increased respect for patient autonomy since an earlier survey in 1985; in most nations more would perform prenatal diagnosis for a couple with 4 daughters who desire a son. A minority (35% in U.S., 14% elsewhere) would perform PND for a deaf couple who want a deaf child, but most (94% in U.S., 62% elsewhere) would do prenatal paternity testing in the absence of rape or incest. About half (51%) would support a woman with fragile X who wants children. The trend to respect patient autonomy was greatest in the U.S. and was least evident in China and India. In general, responses to these cases illustrate a shift away from population or eugenic concerns to a model of genetics focused on the individual. PMID- 12778942 TI - The reactions on Hugo de Vries's Intracellular pangenesis: the discussion with August Weismann. AB - In 1889 Hugo de Vries published Intracellular Pangenesis in which he formulated his ideas on heredity. The expectations of the impression these ideas would make did not come true and publication was negated or reviewed critically. From the reactions of his Dutch colleagues and the discussion with the famous German zoologist August Weissmann we conclude that the assertion that each cell contains all hereditary material was controversial and even more the claim that characters are inherited independently of each other. De Vries felt that he had to convince his colleagues of the validity of his theory by providing experimental evidence. He established an important research program which resulted in the rediscovery of Mendal's laws and the publication of The Mutation Theory. This article also illustrates some phenomena that go beyond an interesting episode in the development of theories of heredity. It shows that criticism from colleagues can move a researcher so deeply that he feels compelled to set up an extensive research program. Moreover it illustrates that it is not unusual that a creative scientist is only partially willing to take criticism on his theories into account. Last but not least it demonstrates that common opinion on the validity of specific arguments may change in the course of time. PMID- 12778943 TI - In focus. Life after BioethicsLine: a reply to Joyce Plaza. AB - The recent closure and removal of BioethicsLine led many researchers to wonder where to turn for their research needs. Joyce Plaza wrote that the closure is a mistake. In this essay I maintain, contra Plaza, that due to its cross disciplinary nature researchers can find bioethics literature in other databases. In developing the search strategy the researcher needs to consider what the problem is about. If the researcher has a philosophical approach in mind, a wise choice would be to use Philosopher's Index; a legal approach suggests using Academic Universe, Westlaw, Lexis or legal databases freely available through state and federal websites. Further, in so far as the National Library of Medicine is integrating citations in BioethicsLine into the NLM Gateway databases (PubMed and LOCATORplus) I point out suggestions on using the latter databases effectively. There is a wealth of information readily available and researchers have much to learn by trying the alternatives. PMID- 12778944 TI - In focus. The brief career of a government advisory committee: one members's perspective. The life and death of the National Human Research Protections Advisory Committee (NHRPAC). PMID- 12778945 TI - In praise of reading. PMID- 12778946 TI - Promoting clinical laboratory science. PMID- 12778947 TI - Waived testing. PMID- 12778948 TI - Bioterrorism: What? Why? and Who? AB - The former Secretary of the Department of Health and Human Services, Donna Shalala, indicated in an address in 1999 that complacency needs to be replaced with a sense of urgency in order for us to deal successfully with the threats of bioterrorism. The attack on September 11, 2001 and the anthrax threats have made our vulnerability clear. We are now living in a new and frightening world. Our complacency is gone. The victims and the survivors shall remain forever in our minds. Dr. Jeffery Koplan, Director, Centers for Disease Control and Prevention in his broadcast, Building Infrastruture to Protect the Public Health said we must look at preparedness in a new way. We need to: build a solid public health infrastructure with grant monies; rapidly address the problem of inadequately trained staff; and address the capacity of a laboratory to produce timely and accurate results for the diagnosis of agents in the investigation of outbreaks. We must take action to prepare the healthcare system to rapidly meet any challenge, overt or covert, that may emerge. PMID- 12778949 TI - Using the urine dipstick to screen out unnecessary urine cultures: implementation at one facility. AB - This study examined the utility of performing urine cultures on biochemically negative urine specimens and details the implementation of a policy to cancel these cultures. Four reactions of the Multistix SG (Bayer, Elkhart IN) urine dipstick (protein, occult blood, leukocyte esterase, and nitrite) were used as biochemical indicators. A three-month retrospective study examining the results of 843 urinalysis/ urine culture pairs indicated that one-third of these cultures were probably unnecessary (negative dipstick/negative culture). Based on these results, a policy was implemented to screen those urine samples having both a urinalysis and urine culture ordered. Over a six-month period, 6,192 urine specimens were evaluated. Of these, 36% (2,260 cultures) were cancelled. Of the 3,932 samples cultured 22.4% (883) were true positives (positive dipstick/positive culture) while 31.6% (1245) had a positive dipstick but grew organisms considered contaminants. The false positive rate was 40% (positive dipstick/negative culture), and the false negative rate was 6%. Implementation of this policy reduced the number of urines cultured by 36%. PMID- 12778950 TI - Differentiating the dermatophytes. PMID- 12778951 TI - The incidence of group B streptococcus in the vaginal tracts of pregnant women in central Alabama. AB - Group B streptococcus (Streptococcus agalactiae) or GBS is the most common cause of neonatal sepsis and meningitis in the United States. One important risk factor for infants who acquire GBS is maternal colonization. Colonization rates have been estimated in various studies to be between 15% and 35% of pregnant women. Colonization rates for black women have also been shown to be higher than for non blacks. Local data were collected and compared to those of other studies. Of the pregnant women in this study overall, 22.76% tested positive for GBS. Black women were colonized by GBS (35.71%) significantly more often than non-black women (19.84%; G = 8.9729, p < 0.00274). Generalized linear models were used to examine age and race. Both maternal age and the interaction of maternal race and age as predictors of infection were ruled out, leaving only race as a significant predictor of colonization. PMID- 12778952 TI - Segmented neutrophil size and platelet morphology in HIV/AIDS patients. AB - OBJECTIVE: A study was conducted to determine if HIV/AIDS patients have smaller than normal size neutrophils and increased prevalence of abnormal platelet morphology. DESIGN: Wright's-stained peripheral blood smears from 100 HIV/ AIDS patients were evaluated for size of segmented neutrophils and degree of abnormal platelet morphology. SETTING: East Carolina University, Greenville, North Carolina. PARTICIPANTS: The study subjects consisted of 100 HIV/AIDS patients seen in an outpatient clinic in a teaching hospital in an academic health center. Peripheral blood smears were made from EDTA tubes drawn as a part of a routine immunology panel. MAIN OUTCOME MEASURES: Segmented neutrophils from each of ten oil immersion fields were measured for diameter with a micrometer and the average diameter calculated. In addition, any platelet morphology abnormality, which was noted in at least five oil immersion fields, was recorded. One researcher evaluated one slide on each patient, and the second researcher randomly selected 20% of the subjects and performed the same procedure on a second slide for quality assurance of results. RESULTS: A segmented neutrophil mean diameter of 15.1 microns was found. Though this mean is a mere 0.1 microns above the upper limit of the normal range of 10 to 15 microns, 53% of the patients had an observed average diameter above 15 microns. The HIV/AIDS patients' mean diameter was statistically different when compared to a normal population mean of 12.0 microns (T-test = 16.15, p < .0001), thus, showing a tendency for HIV/AIDS patients' segmented neutrophils to be large. Over half of the study subjects demonstrated abnormal platelet morphology including agranularity, small size, and giant size. CONCLUSION: Neutrophil size as based on cell diameter, was found to be significantly larger for a sample of HIV/AIDS patients than the normal mean size. There was also a tendency for platelet morphology to be abnormal. PMID- 12778953 TI - The learning and application of generic skills by CLSs/MTs who have 'left the field'. AB - OBJECTIVE: To determine whether generic skills that dinical laboratory scientists (CLSs)/medical technologists (MTs) learned as students and/or practitioners are applied to jobs outside the field of CLS/MT; and to determine if there are any significant differences in learning and/or doing these skills by CLS/MT majors vs. non-CLS/MT majors. DESIGN: An Occupational Change Survey was sent to CLS/MT practitioners who had identified themselves as having left the field (LTF) of CLS/MT. The participants were asked whether or not they were CLS/MT majors as undergraduates, whether they utilized generic baccalaureate level skills in their LTF jobs, and whether or not they learned these skills as CLS/MT students and/or practitioners. The skills were: problem solving, decision making, troubleshooting, analytical reasoning, data correlation, precision studies, quality assessment, teaching, research, communication, technical writing, computer use, utilization review, and supervision. SETTINGS AND PARTICIPANTS: The survey was sent to 105 participants of an ongoing longitudinal study who identified themselves as having LTF. MAIN OUTCOME MEASURES: Responses for doing/utilizing the skills were grouped as 'Yes' if participants indicated they frequently or sometimes used the skills in their LTF jobs, and 'No' if they indicated they rarely or never used the skills in their LTF jobs. Responses for learning the skills were grouped as 'Yes' if participant indicated they learned the skills as CLS/MT students, practitioners or both and 'No' if they indicated they never learned the skills as CLS/MT students, practitioners, or both. Participants indicated whether or not they were CLS/MT majors in college. Chi square analyses were performed to test for any statistical significant (p = 0.05) differences between: doing and learning the skills, doing the skills and being a CLS/MT major, and learning the skills and being a CLS/MT major. RESULTS: The response rate for the survey was 48% (50/103). Chi square analyses could not be performed for doing the skills in the LTF jobs for three variables: problem solving, analytical reasoning, and computer use because all respondents reported that they used these skills. Chi square analyses indicated there were no significant differences between doing and learning the skills in the LTF job for the entire sample group for all remaining skills except supervision. There were no significant differences between doing the skills in the LTF job and being a CLS/MT major. A statistically significant difference in learning the skills was observed between CLS/MT majors and non-CLS/MT majors for the following skills: problem solving, correlating data, precision studies, research, analytical reasoning, and troubleshooting. The 'Yes' answer frequencies for learning the skills was higher for the CLS/ MT majors for all the generic skills except teaching, where they were equal, and utilization studies where they were lower. CONCLUSION: The results indicate that, in general, for this sample group, generic skills learned as CLS/MT students and/or practitioners can be and are applied to a wide variety of LTF jobs. Furthermore, CLS/MT majors learned the generic skills at least as well, if not better, than other baccalaureate level laboratory practitioners who obtained degrees in other areas. PMID- 12778954 TI - The minimum concentration of fibrinogen needed for platelet aggregation using ADP. AB - OBJECTIVE: Determine the minimum concentration of plasma fibrinogen needed to stimulate the aggregation of platelets, collected from normal subjects, using ADP. DESIGN: Platelet rich plasmas (300 x 10(9) platelets/L) were made and adjusted to final fibrinogen concentrations of 75, 19, 5, and 0 mg/dL using fibrinogen free serum. Each fibrinogen concentration in all twelve subjects was aggregated with ADP SETTING: Research laboratory in the Department of Clinical Laboratory Science at Saint Louis University. PARTICIPANTS: Twelve healthy volunteers of both genders, between the ages of 18 and 60 years who were not pregnant and weighed at least 110 pounds were included in the study. Subjects were excluded from the study if they had ingested aspirin within one week prior to blood collection. In addition, subjects with a history of bleeding disorders such as afibrinogenemia, hypofibrinogenemia, von Willebrand disease, and Bernand Soulier disease were rejected from the study. MAIN OUTCOME MEASURES: Platelet aggregation tracings were analyzed for amplitude and compared across plasma fibrinogen concentrations. In addition, the type of curve (monophasic vs. biphasic), smoothness and aggregation stability were also noted. RESULTS: The results show that aggregation occurred with every dilution of fibrinogen tested and that the amplitude of the aggregation curves appears not to be dependent on plasma fibrinogen. CONCLUSIONS: The results indicate that platelets from healthy individuals previously exposed to normal fibrinogen levels will aggregate equally well in decreasing plasma fibrinogen concentrations and even in the absence of plasma fibrinogen using ADP as the aggregator. PMID- 12778955 TI - Malignancy: an evolving definition of a cancer cell. PMID- 12778956 TI - The new WHO nomenclature: introduction and myeloid neoplasms. PMID- 12778957 TI - The new WHO nomenclature: lymphoid neoplasms. AB - The development of the WHO classification of lymphoid neoplasms is a remarkable example of cooperation and communication between pathologists and oncologists from around the world. Joint classification committees of the major hematopathology societies will periodically review and update this classification, facilitating further progress in the understanding and treatment of hematologic malignancies. PMID- 12778958 TI - Laboratory services payment threatened once again. PMID- 12778959 TI - Mycology at a distance. AB - GermWare Mycology is an image-rich, CD-ROM-based instruction divided into tutorial and reference programs. The tutorial program, designed for new students, provides only for sequential progress through each of the subject modules, so that each page of information is seen. In contrast, the reference program allows the more experienced learner with random and direct access to each facet of information. The aspergilli, the agents of chromomycosis, the dermatophytes, the dimorphic fungi, the hyaline molds, the dematiaceous molds, the yeasts, and the zygomyces are divided into separate modules. The tutorial program also includes an opening 'isolation procedures' module, in which details of specimen collection, culture media, and microscopic techniques are presented. The random access program includes system maps separating out each of the fungal species, and flow diagrams allowing an algorithm approach to species identifications. A global map is also included through which each fungal species can be directly accessed by the simple click of the mouse. Random access to information on the ecology, clinical presentations, pathology and therapy of the various mycotic diseases is also a feature of the reference program. A series of self-assessment exercises is included at the end of each module, with immediate 'pop-up' feedback to both correct and incorrect answers. The entire program includes over 2500 screens and over 700 color images and diagrams. GermWare Mycology is available through the Colorado Association for Continuing Medical Laboratory Education (CACMLE), who also can provide continuing education credits for individuals who complete a separate examination. For more information contact CACMLE at (303) 321 1734 or info@cacmle.org. PMID- 12778960 TI - Antibody identification in a patient with chronic lymphocytic leukemia: a case study. PMID- 12778961 TI - Diagnosing hemolytic disease of the newborn. AB - A Rh negative, pregnant female presented to a major medical center for possible Rh alloimmunization. This female had nine previous pregnancies, including three spontaneous abortions, four live births, and two fetal demises. Because of poor prenatal care, the immunization Rh immune globulin was administered to only the first two pregnancies. After much laboratory testing and treatment, this tenth pregnancy also ended in fetal demise. PMID- 12778962 TI - Genetics and molecular diagnostics in the clinical laboratory science curriculum. AB - OBJECTIVE: To determine the nature and extent of education in human genetics and molecular diagnostics in clinical laboratory science (CLS) programs throughout the U.S. DESIGN: A written survey was mailed to 263 CLS programs. Data were expressed as raw numbers and percentages of responses. SETTING: State University of New York, Upstate Medical University. PARTICIPANTS: There were 162 responses and 151 usable surveys. Most respondents (86.8%) were department chairs/CLS program directors; 13.2% were CLS faculty or educational coordinators. MAIN OUTCOME MEASURES: Questions were designed to determine frequency of CLS programs providing education in genetics, specific molecular methods and clinical applications, format of instruction, satisfaction levels with education provided, and perceptions on importance of teaching genetics, molecular diagnostics, and related hands-on experiences. RESULTS: Over 92% of CLS programs teach human genetics and molecular diagnostics in varied formats. Polymerase chain reaction was the most frequently taught molecular method; microorganism detection, the most commonly taught clinical application. More programs teach theory than provide hands-on experience in molecular diagnostics. Only 59 (39.1%) teach related ethical issues. Sixty-seven respondents (44.4%) were dissatisfied with the education they provide, due to lack of time to teach the material (n = 49; 73.1%), lack of knowledgeable faculty (n = 43; 64.2%), and expense of methods (n = 37; 55.2%). Most respondents felt it was important to include human genetics (n = 145; 96%) and molecular diagnostics (n = 149; 98.7%) in their curriculum, and related hands-on experiences in the student laboratory (n = 106; 70.2%) or clinical rotation (n = 135; 89.4%). Over 82% (n = 124) expected instruction of molecular diagnostics to increase in the next five years. CONCLUSION: Most CLS programs include human genetics and molecular diagnostics in their curriculum, and expect the education they provide to increase in the next 5 years. In order to meet this expectation, CLS programs may need to provide opportunities for faculty training, seek funding to cover the cost of methods, and consider innovative curriculum changes. PMID- 12778963 TI - An entry-level MS degree in clinical laboratory science: is it time? AB - OBJECTIVE: The study was undertaken to address the following questions: 1) Does the scope of practice of the clinical laboratory scientist require an entry-level master's (MS) degree? 2) How would a change to an entry-level MS degree in clinical laboratory science (CLS) affect educational programs, the practice field, and students? and 3) Based on this study, what recommendations can be made to CLS educators? DESIGN: Surveys were developed to assess the opinions of educators, managers, and practitioners on the need for an entry-level MS degree in CLS. Surveys were also sent to students to assess their interest in an entry level MS degree and their perceptions of the advantages and disadvantages of this type of program. Surveys sent to educators included questions addressing the effect of a change to an entry-level MS degree in CLS on enrollment and program viability. Managers were asked questions concerning job expectations and compensation for graduates with an entry-level MS degree and practitioners were asked about their interest in this type of program. PARTICIPANTS: The sample for the survey included 280 directors of National Accrediting Agency for Clinical Laboratory Sciences (NAACLS) educational programs, 600 managers randomly selected from the Clinical Laboratory Management Association (CLMA) mailing list, 600 practitioners randomly selected from the American Society for Clinical Laboratory Science (ASCLS) mailing list, and 1400 CLS students selected by program directors. MAIN OUTCOME MEASURES: Educators, managers, and practitioners were asked to read 12 statements related to educational preparation for entry into CLS and indicate their level of agreement on a five point scale. Mean responses to these questions were compared for educators, managers, and practitioners, for educators in hospital-based and university-based programs, and for managers with BS and advanced degrees. Responses to demographic and other forced-choice type questions related to entry-level MS programs were counted and reported. RESULTS: Response rates of 58% (educators), 28% (practitioners), 39% (managers), and 40% (students) were obtained. Educators, managers, and practitioners all agreed that the scope of practice of CLS does not require an entry-level MS degree and that the MS degree is appropriate for those practitioners who wish to further their education. There were no major differences in educators', managers', and practitioners' responses to questions on the need for an MS in CLS. Students indicated that they would be interested in an entry-level MS program if the additional education would give them higher salaries and more job opportunities. Students who entered their CLS program with a baccalaureate (BS) degree were more interested in the entry-level MS option than students who entered with an associate degree or high school diploma. Managers indicated that they would not pay a graduate with an entry-level MS degree more than a graduate with a baccalaureate degree. CONCLUSION: There is currently no support for an overall change from the BS degree to the MS degree as the entry-level requirement for CLS practitioners. Entry-level MS programs in CLS may be attractive to students who already have BS degrees. PMID- 12778964 TI - The Laboratory Response Network for bioterrorism. AB - OBJECTIVE: To describe the function and levels of analysis performed by members of the Laboratory Response Network in coping with biological agents of terrorism. DATA SOURCES: Current literature and the Internet. CONCLUSIONS: The Laboratory Response Network is designed to enable rapid, safe, and accurate diagnosis of disease in order to mobilize the nation's response to acts of bioterrorism. PMID- 12778965 TI - Anthrax 2001--lessons learned: clinical laboratory and beyond. AB - OBJECTIVE: Re-visit the 2001 anthrax outbreak to assess the ideas and concepts learned from the event as they relate to the illness and to bioterrorism preparedness. DATA SOURCES: Current literature. CONCLUSION: A multitude of lessons have been brought to light. The future of bioterrorism preparedness depends on whether those lessons are acknowledged and acted upon. PMID- 12778966 TI - Anthrax 2001--lessons learned by public health laboratories. AB - OBJECTIVE: To share lessons learned by one local public health department during the anthrax outbreak and associated public hysteria during the autumn of 2001. DATA SOURCES: Current literature and personal experience. CONCLUSIONS: Previous planning for a possible bioterrorism event is essential. Management of the communication and testing process is essential for the protection of the public. PMID- 12778967 TI - 2002 JCAHO laboratory surveys. AB - OBJECTIVE: Review changes for organizations surveyed under the 2002-2003 Comprehensive Accreditation Manual for Pathology and Clinical Laboratory Services published by the Joint Commission on Accreditation of Healthcare Organizations. DATA SOURCES: Current literature, Joint Commission on the Accreditation of Healthcare Organizations. CONCLUSIONS: Few standards changes have been made in the manual, but the Joint Commission on the Accreditation of Healthcare Organizations has refined the survey process in subtle ways and new requirements in the area of proficiency testing are in place. PMID- 12778968 TI - Generating recombinant anti-idiotypic antibodies for the detection of haptens in solution. AB - A new method is described for generating recombinant human and chicken antibody fragments for accurate quantification of haptens in solution. The chemistry of labelling small molecules has always been a problem in the development of immunoassays. Here, we describe a specific panning procedure that enables the selection of recombinant anti-idiotypic phage antibodies that bind to hapten binding molecules (e.g., antibodies) in the absence of the hapten, but are displaced in a highly specific and concentration dependent manner, in the presence of the hapten. The major advantage of such a detection system is that there is no need to label the hapten or to covalently attach it to a solid phase. In this study we demonstrate, using cortisol and aldosterone as model haptens, that the recombinant antibody phage display technology offers great possibilities to generate recombinant anti-idiotypic antibodies. Furthermore, we show that such antibodies can be used successfully to design highly sensitive immunoassays for the quantification of small molecules. PMID- 12778969 TI - Use of defined estrone glucuronide-hen egg white lysozyme conjugates as signal generators in homogeneous enzyme immunoassays for urinary estrone glucuronide. AB - Three structurally characterized estrone glucuronide-lysozyme conjugates, E1 (a 60:40 mixture acylated at K3 and K97), E3 (acylated at K33), and E5 (acylated at both K33 and K97) were isolated and purified using a combination of cation exchange chromatography on S-sepaharose in 7M urea and hydrophobic interaction chromatography on butyl sepharose. Urea was essential to separate the conjugates into six chromatographically homogeneous fractions. In the absence of urea, complex mixtures of lysozyme and the six conjugate fractions were always encountered. The E1, E3, and E5 conjugates were highly inhibited by a sheep polyclonal anti-estrone glucuronide antibody only after the hydrophobic interaction chromatography step. The high level of inhibition enabled all three conjugates to be utilized as signal generators in homogenous enzyme immunoassays for urinary estrone glucuronide. Despite the apparently higher affinity of E3 for the antibody, both E1 and E3 gave standard curves that were indistinguishable provided that 1.7-fold more antiserum was used for E1. Both E1 and E3 yielded menstrual cycle urinary data that agreed with that provided by the Ovarian Monitor pre-coated assay tubes. Although, the menstrual cycle pattern was similar for the three signal generators, the E1G excretion rates yielded by E5 as the signal generator were only 60% of the reference values. Despite structural differences, there was no advantage gained in separating E1 and E3, but higher substituted conjugates such as E5 need removal for best assay performance. PMID- 12778970 TI - Comparative studies with penicillinase, horseradish peroxidase, and alkaline phosphatase as enzyme labels in developing enzyme immunoassay of cortisol. AB - Relative merit of different enzyme labels for measuring cortisol directly in serum by competitive enzyme immunoassay (ELISA) was examined. Cortisol-21 hemisuccinate was labeled separately with penicillinase, horseradish peroxidase (HRP), and alkaline phosphatase (ALP) under identical reaction conditions. Antibody developed in rabbits against cortisol-3-0-(carboxymethyl)-oxime-bovine serum albumin was used to coat polystyrene tubes that were precoated with anti rabbit gamma globulin (ARGG). Cortisol standards were prepared in steroid-free human serum in buffer (1:4) contaning 8-anilino-1-naphthalene sulfonic acid (8 ANS). Assay buffer also consisted 8-ANS. The assay involved adding standard cortisol or serum sample to antibody-coated tubes, followed by addition of enzyme label and buffer, and incubation for 2 h at 37 degrees C. The whole procedure took 3 h for completion. All three labels proved to be sensitive, with a slope around -2.0. Although penicillinase as an enzyme label was highly sensitive and stable compared with others, the assays were not always accurate and precise, especially at low concentrations of cortisol. This was mainly due to the color reagent used for measuring penicillinase activity. Serum samples that underwent 2 3 freeze-thaw cycles gave high values with HRP label compared with ALP. Therefore, utilizing ALP as an enzyme label, an ELISA was developed and its performance was comparable with some of the commercial kits already in the market. PMID- 12778971 TI - Substitution of carbonate buffer by water for IgG immobilization in enzyme linked immunosorbent assay. AB - The first step of enzyme linked immunosorbent assay (ELISA), namely, adsorption of antigen or antibody to the plastic microtiter well plate, was studied as a function of insolubility of IgG in water. Immobilization efficiency was assessed in terms of number of wells coated per milliliter of primary antiserum. We have compared different coating/immobilization protocols, i.e., direct and indirect immobilization of primary antibody to the plastic microtiter well plate using carbonate buffer and phosphate buffer with glutaraldehyde. We have observed efficient coating when the immobilization of primary antibody through an immunobridge technique was performed, where water was used as a coating medium. It gave a higher number of wells coated per milliliter of anti-serum (primary or secondary) than other compared coating protocols and it allowed the use of serum (non-immune) and anti-serum (primary and secondary antibody) dilutions, avoiding the need for gamma-globulin purification from normal and immunized serum. PMID- 12778972 TI - One step enzyme linked immunosorbent assay for direct estimation of serum testosterone. AB - One step competitive enzyme linked immunosorbent assay (ELISA) for direct estimation of testosterone in human serum is described. Testosterone-3-O carboxymethyl-oxime-bovine serum albumin (testosterone-3-O-CMO-BSA), was used as immunogen and testosterone-3-O-carboxymethyl-oxime-adipic-acid dihydrazide horseradish peroxidase (testosterone-3-O-CMO-ADH-HRP) was used as tracer. To the testosterone antibody coated microtiter wells, standard or serum samples (100 microL), along with testosterone-3-O-CMO-ADH-HRP conjugate (100 microL) were incubated for 1 h at 37 degrees C. Bound enzyme activity was measured by using tetra methyl benzidine/hydrogen peroxide (TMB/H2O2) as a substrate. In this new strategy, charcoal stripped pooled human serum spiked with non-cross reactive C18, C19, C21, and C27 steroids, used for preparing the standards and blocking the sex hormone binding globulin (SHBG)/and other steroid binding globulins (SBG). The sensitivity of the assay was 0.015 ng/mL. The intra-assay and inter assay coefficients of variation (CVs) were ranged from 7.8 to 11.8 and 4.8 to 10.4, respectively. The serum testosterone values, obtained by this method, were correlated well with those obtained by radioimmunoassay r = .98 (n = 100). PMID- 12778973 TI - Development and application of a pig IL-8 ELISA detection system. AB - Interleukin 8 (IL-8) is a chemotactic and activating chemokine, especially for neutrophils, which plays an important role in inflammatory process. A pig IL-8 specific enzyme-linked immunosorbent assay (ELISA) was developed to measure IL-8 concentrations in cell culture supernatants and biological fluids. A streptavidin biotin amplified sandwich method uses mouse capture mAb IZ8.03 and detection biotinylated mouse mAb IZ8.04 against recombinant pig IL-8. The assay specifically and reproducibly recognizes both recombinant and natural pig IL-8. A working range of the assay is 16-1000 pg/mL and takes a mere 3.5 h of incubation time. This pig IL-8 ELISA is a suitable alternative way of measurement of IL-8 concentrations to time consuming and laborious IL-8 bioassays. PMID- 12778974 TI - Staff members' feelings toward psychiatric patients related to their own and the patient's self-image and gender. AB - The relation between staff members' feelings toward a patient and their own and the patient's self-image in different gender combination groups was studied. Staff at 16 psychiatric treatment homes for patients with severe psychopathology reported their feelings toward their patients on a number of occasions. At the start of treatment, both staff members and patients rated their self-images using the Structural Analysis of Social Behavior (SASB). Male staff seemed less influenced by the patient, with their feelings relating mainly to aspects of their own self-image, while the feelings of the female staff were more related to the patient's self-image. The patient's diagnosis was less important for a staff member's feelings than that member's self-image. Generally, the relation between feelings and self-image was stronger for negative feelings. The results point to the importance of understanding more about the influence of staff members' self structure on their negative feelings toward their patients and how this relates to both the staff member's and patient's gender. PMID- 12778975 TI - The role of locomotion in the acquisition and transfer of spatial knowledge in children. AB - The role of locomotion in the acquisition and transfer of spatial knowledge was investigated in 144 five-, seven- and eleven-year-old children. Two experiments were conducted in the Kiel locomotor maze. In the first experiment, one group of children explored the spatial layout by walking through the maze, while another learned the maze by surveying the layout. In the second experiment, children were exposed to one of two orientation tests in the maze, one of which could be solved using "landmark orientation", the other only using a "relational place orientation". Children sitting by the side of the experimental chamber surveying the maze needed fewer trials to learn the spatial layout than children exploring the environment in the locomotion condition, but in the orientation test demanding the "relational place orientation" children who had explored the maze in the locomotion condition outperformed the children in the non-locomotion condition. Results are discussed in the context of cognitive mapping models. PMID- 12778976 TI - Measuring implementation intentions in the context of the theory of planned behavior. AB - The usefulness of measuring implementation intentions in the context of the theory of planned behavior (TPB) was explored among 112 Norwegian college students. They responded to a questionnaire measuring past behavior, perceived behavioral control, behavioral intentions, implementation intentions, and actual performance of regular exercising and recycling of drinking cartons. Implementation intentions were measured using five items relating to recycling and four items relating to exercise, which showed satisfactory internal consistencies. Consistent with the main prediction, the presence of implementation intentions was related to performing the two behaviors, although behavioral intentions were the strongest determinant for both behaviors. The results suggest that the TPB may benefit from inclusion of the concept of implementation intentions to provide a more complete understanding of the psychological process in which motivation is translated into action. PMID- 12778977 TI - Genetic and environmental causes of the interrelationships between self-reported fears. A study of a non-clinical sample of Norwegian identical twins and their families. AB - The present study reports results from a study of the self-reported fears of identical twins and their spouses and offspring. Factor analysis with oblique rotation of questionnaire responses yielded four correlated fear dimensions: situational fears, illness-injury fears, social fears, and fear of small animals. Models allowing for genetic and cultural transmission, together with specially correlated environments for twins, were fitted, both for separate fears and across fears. Simple models with only genetic and uncorrelated environments were sufficient to account for each the fear dimensions considered separately. The cross-dimensional analyses revealed a genetic and an environmental factor common to the four fear dimensions, together with fear-specific genetic and environmental factors. The impact of the common genetic and common environmental factor varied across dimensions. No evidence of cultural transmission or specially correlated twin environments of the cross-dimensional environments was detected. It is concluded that both common and fear-specific genes and (individual-specific) common and fear-specific environments are necessary to account for the data. The results are discussed in terms of the prepared learning hypothesis and the expectancy bias hypothesis. PMID- 12778978 TI - Classifying homicide offenders and predicting their characteristics from crime scene behavior. AB - A theoretical distinction between instrumental and expressive aggression was used in analyzing offender characteristics and their associations with crime scene actions in Finnish homicides. Twenty-one variables reflecting the offenders' criminal activity, previous relationships with intimates and victims, and general social and psychological adjustment were derived from files of single offender/single-victim homicides occurring between 1980 and 1994 (n = 502). Additionally, three variables describing post-offense actions and police interview behavior were included. A multidimensional scaling procedure was used to investigate the interrelationships between the variables. A distinction between expressive and instrumental characteristics was observable in the empirical structure, which was divided into three subthemes of Instrumental, Expressive: Blood, and Expressive: Intimate. Associations between the characteristics with five previously identified subthemes of crime scene actions were computed. In addition, the subthemes of crime scene actions were related to post-offense actions and police interview behavior, with Expressive themes being associated with less denial as well as a greater likelihood of surrendering and confession. The practical usefulness for police investigations and theoretical implications of the results are discussed. PMID- 12778979 TI - Early phonological skills as a predictor of reading acquisition: a follow-up study from kindergarten to the middle of grade 2. AB - The purpose of this study was to investigate the power of early measures of phonological skills (phonemic awareness, rapid naming, short-term memory) in predicting later reading skills at various points of time. About 70 children were followed from the end of kindergarten to the middle of grade 2. Correlation analyses were performed as well as a linear growth curve analyses. In the traditional regression analysis, phonemic awareness in kindergarten explained about 27% of the variance in word reading six months later and about 9.5% of the variance at the end of grade 1. Even when prior level of reading skill was included in the predictive equation, a significant amount of variance was still explained by phonemic awareness. The other predictor variables did not explain any variance in word reading, and phonemic awareness did not predict any variance in reading skills in grade 2. When using sentence reading as the dependent variable, phonemic awareness explained about 16% of unique variance after six months, and about 13% of the variance in the middle of grade 2. Similarly, when employing growth curve analysis, phonemic awareness was the only phonological factor that accounted for significant variance in the word reading slope, explaining about 25% of its variance, whereas naming and short-term memory did not explain any unique variance. The lack of predictive power of phonemic awareness on the sentence b-slope is assumed to be caused by unreliable sentence scores in kindergarten. PMID- 12778980 TI - Gender bias in the evaluation of new age music. AB - Eminent composers in Western European art music continue to be predominantly male and eminence in contemporary pop music is similarly male dominated. One contributing factor may be the continuing under-valuation of women's music. Possible anti-female bias in a contemporary genre was investigated using the Goldberg paradigm to elicit judgments of New Age compositions. Since stronger stereotyping effects occur when information provided about individuals is sparse, fictitious male and female composers were presented either by name only or by name with a brief biography. Evidence for anti-female bias was found in the name only condition and was stronger when liking for the music was controlled. Other findings were the tendency for females to give higher ratings, and the association of gender differences in liking of the music with ratings of quality in the name-only condition. These results are relevant to the design of formal assessment procedures for musical composition. PMID- 12778981 TI - Rorschach administration: a comparison of the effect of two instructions given to an inpatient sample of drug addicts. AB - The effect of administering the Rorschach Inkblot Method under two instructional sets was compared on three classes of outcome variables: the frequency with which subjects asked questions about the test; the frequency of brief protocols (fewer than 14 responses); and 17 traditional Rorschach structural summary scores. Sixty subjects, obtained from three inpatient psychiatric clinics treating drug addicts, randomly received either the short pre-testing instruction "What might this be?" originally developed by Herman Rorschach and recommended in the Comprehensive System, or a longer and more elaborated instruction, which for many years has been the standard instruction in Norway. Compared with the Norwegian instruction, the short instruction produced significantly more questions to the examiner about the test. For the other outcome measures no differences were observed. PMID- 12778982 TI - Models-of-self and models-of-others as related to facial muscle reactions at different levels of cognitive control. AB - The hypotheses of this investigation were based on attachment theory and Bowlby's conception of "internal working models", supposed to consist of one mainly emotional (model-of-self) and one more conscious cognitive structure (model-of others), which are assumed to operate at different temporal stages of information processing. Facial muscle reactions in individuals with positive versus negative internal working models were compared at different stages of information processing. The Relationship Scale Questionnaire (RSQ) was used to categorize subjects into positive or negative model-of-self and model-of-others and the State-Trait Anxiety Inventory was used to measure trait anxiety (STAI-T). Pictures of happy and angry faces followed by backward masking stimuli were exposed to 61 subjects at three different exposure times (17 ms, 56 ms, 2,350 ms) in order to elicit reactions first at an automatic level and then consecutively at more cognitively elaborated levels. Facial muscle reactions were recorded by electromyography (EMG), a higher corrugator activity representing more negative emotions and a higher zygomaticus activity more positive emotions. In line with the hypothesis, subjects with a negative model-of-self scored significantly higher on STAI-T than subjects with a positive model-of-self. They also showed an overall stronger corrugator than zygomatic activity, giving further evidence of a negative tonic affective state. At the longest exposure time (2,350 ms), representing emotionally regulated responses, negative model-of-self subjects showed a significantly stronger corrugator response and reported more negative feelings than subjects with a positive model-of-self. These results supported the hypothesis that subjects with a negative model-of-self would show difficulties in self-regulation of negative affect. In line with expectations, model-of-others, assumed to represent mainly knowledge structures, did not interact with the physiological emotional measures employed, facial muscle reactions or tonic affective state. PMID- 12778983 TI - School-related stress, social support, and distress: prospective analysis of reciprocal and multilevel relationships. AB - This three-wave prospective study investigated the reciprocal relationships among school-related stress, school-related social support, and distress in a cohort of 767 secondary school students (mean age 13.9 years). Stress, support, and distress were measured at three occasions with six-month lags between. Reciprocal relationships were analyzed with multivariate multilevel modeling (MLwiN). Each of the three factors at baseline predicted change in one or two of the other factors at subsequent measurements, indicating a complex pattern of reciprocal relationships among stress, support, and distress across time. A high level of distress at baseline predicted a lower level of support and a higher level of stress six months later. High levels of stress at baseline predicted a higher level of distress and a lower level of support 12 months later. The results are consistent with a transactional and dynamic model of stress, support, and distress, and indicate the need to view school-related stress, support, and distress as mutually dependent factors. PMID- 12778984 TI - Preattentive processing of alcohol stimuli. AB - An experiment was conducted to test the automatic analysis of briefly presented alcohol stimuli in alcohol-dependent individuals. Alcoholics and controls were exposed to four different conditions: two brief (30 ms) and two long (130 ms) exposure conditions, each containing alcoholic and non-alcoholic pictures. Heart rate (HR) interbeat intervals were recorded and phasic cardiac responses assessed. Alcoholics had a stronger initial HR deceleration after exposure to masked alcohol slides compared with masked control slides, indicating a preattentive analysis of alcohol stimuli. This initial HR deceleration in the masked condition suggests an automatic attentional focusing to degraded alcohol cues. No such attentional effect was found when the pictures were presented unmasked and were clearly perceived. The implication of these results for the understanding of relapse in addictive behavior is discussed. PMID- 12778985 TI - ALLHAT. Is it all hat? PMID- 12778986 TI - Ensuring the integrity of medicine. PMID- 12778987 TI - ECG of the month. Premature beats. Ventricular parasystole. PMID- 12778988 TI - Radiology case of the month. Raccoon eyes and abdominal distention. Neuroblastoma. PMID- 12778989 TI - Pathology case of the month. Abdominal pain in pregnancy. HELLP syndrome with subcapsular hematoma of the liver. PMID- 12778990 TI - The New Orleans Journal of Medicine: April 1868. PMID- 12778991 TI - Clinical case of the month. Weakness in an 88-year-old man. Multiple myeloma. AB - Multiple myeloma is a malignant proliferation of plasma cells that typically occurs in older individuals and is characterized by anemia, renal failure, bone pain, lytic lesions, and a monoclonal gammopathy. This disease is associated with significant morbidity and mortality. PMID- 12778992 TI - Mitral stenosis: I. Anatomical, physiological, and clinical considerations. AB - Because of the dramatic decline in the incidence of acute rheumatic fever in the United States and much of the developed world over the past 70 years, mitral stenosis, once a common valvular problem, is now distinctly uncommon in many countries. Nevertheless, because mitral stenosis maintains a high prevalence in developing countries and among emigrants from those countries to the United States, continued awareness of the condition is warranted. Furthermore, failure to recognize mitral stenosis not only precludes many effective therapies but in so doing may result in serious complications. PMID- 12778993 TI - MRI-documented spontaneous regression of cervical disc herniation: a case report and review of the literature. AB - Spontaneous regression of the herniated lumbar discs is a well-established phenomenon. This phenomenon of disappearance of the disc has been reported occasionally in the cervical spine and very rarely in the thoracic regions. In this article, we report the rare occurrence of spontaneous resolution of one large herniated cervical disc (C6-7) with simultaneous progression of another disc prolapse (C4-5) in a 46-year-old man. PMID- 12778994 TI - Chart review and pilot study of blood pressure control in acute ischemic stroke. AB - BACKGROUND: There are no data describing a relation between the course of blood pressure and outcome in acute ischemic stroke. We found such a link in a chart review, and then used that data to design a pilot study. We report on the chart review and the first 13 patients enrolled. METHODS: We examined charts of patients admitted to our service in 2000 and stratified them by good (Rankin scale < 4 on discharge, meaning functional independence with a device or better) or poor outcome. A significant difference in blood pressure course and outcome was found, and we then designed a pilot study in an attempt to reproduce these findings. RESULTS: Analysis of the chart review data from 54 patients revealed that the course of mean arterial pressure was different in the two groups: the mean arterial pressure in the group with good outcome decreased about 2 mmHg daily while there was no change in the group with poor outcome (p=0.002 for a difference between slopes). In the pilot trial there were no differences between mean arterial pressure parameters or outcome in either the placebo or treatment groups. Nearly all patients with an admission NIH Stroke Scale score of 6 or less were discharged home. CONCLUSIONS: While our retrospective data were suggestive, we were unable to reproduce findings linking gradual control of blood pressure and outcome in acute ischemic stroke. This pilot study may be used to refine future trials. PMID- 12778995 TI - Genes that permit or prevent infections. AB - In addition to heritable immunodeficiencies and polygenic susceptibility, acquisition genes and resistance genes are important genetic determinants of infectious diseases. Normal forms of acquisition genes permit entry of microorganisms into cells; mutations prevent entry. Normal forms of resistance genes prevent infections; mutations permit infections. The Genome Project has resulted in a growing list of each type. The current lists support important clinical principles. Many, if not most, pathogens readily enter cells unless prevented by antisepsis, blocking, and sterile techniques. Many, if not most, infections occur primarily in persons with heritable mutations of resistance genes. PMID- 12778996 TI - Esophageal duplication cyst. PMID- 12778997 TI - Management of endobronchial hamartoma. AB - Pulmonary hamartomas are uncommon benign tumors with a population incidence of 0.25%. They occur in parenchymal and endobronchial locations, the latter representing 10 to 20% of the total number of pulmonary hamartomas. Parenchymal tumors are generally small (<4 cm) and well-circumscribed and present asymptomatically as an incidental radiographic finding. Endobronchial tumors are generally broad-based lobulated nodules, which grow slowly, resulting in symptoms of airway obstruction including cough, hemoptysis, dyspnea, or obstructive pneumonia. Histologically the tumors consist of varying combinations of benign elements including cartilage, connective tissue, fat, and smooth muscle. We present a 67-year-old man who presented with persistent right lower lobe pneumonia and was found to have a right mainstem endobronchial hamartoma which was resected endoscopically by laser mechanical debridement. A review of the pathobiology of endobronchial hamartoma is presented. PMID- 12778998 TI - Ovarian cancer in women with prior hysterectomy. AB - OBJECTIVE: To determine the proportion of women with ovarian cancer treated at our institution who may have had their disease prevented if oophorectomy had been performed during prior hysterectomy. MATERIALS AND METHODS: One hundred twelve women with ovarian cancer treated at our institution were identified through records in our Gynecologic Oncology office. We identified 19 women (17%) who had undergone prior hysterectomy without bilateral oophorectomy (Group 1). We compared age at cancer diagnosis, stage of disease, and cell type between these women and the 93 without prior hysterectomy (Group 2). RESULTS: The mean age at hysterectomy for women in Group 1 was 33.8+/-5.9 years (range 21-44 years). There were 7 women over the age of 35 years, 3 of whom were over the age of 40 years at time of hysterectomy. Overall, 17.0% of ovarian cancer cases theoretically could have been prevented if bilateral oophorectomy had been performed at the time of hysterectomy. However, this drops to only 2.7% if oophorectomy was limited to women over 40 years. CONCLUSION: Women in our study who underwent prior hysterectomy did so at a relatively young age. This limited the impact prophylactic oophorectomy could have had on ovarian cancer prevention in our population. PMID- 12779000 TI - Human genome sequence completed. PMID- 12778999 TI - Neurological diseases and RNA-directed gene regulation: prospects for new diagnostics and therapy. PMID- 12779001 TI - SARS: new sequence could lead to better diagnostic tests. PMID- 12779002 TI - Nanogen introduce new products for detecting genetic mutations. PMID- 12779003 TI - Roche PCR-based test enhances ability to measure viral loads of HIV patients. PMID- 12779004 TI - Aspects of molecular diagnostics and therapy in obstetrics and gynecology. AB - Scientific progress and information relating to the theoretical and clinical work being carried out in the field of obstetrics and gynecology has dramatically increased due to recent developments in molecular biology. Molecular obstetrics and gynecology is therefore the link between the different sections in obstetrics and gynecology. At present, the molecular understanding of cellular pathways is much greater than that of the direct integration of molecular diagnostics and therapy in routine clinical practice. The use of molecular diagnostics, such as preimplantation diagnostics or predictive genetic testing, still has technical problems as well as novel, and to date unclear, social, ethical and legal implications. To date, the technical elements of molecular therapy have not yet fulfilled their expectations. In the broad spectrum of obstetrics and gynecology, new molecular discoveries are influenced not only by technical but also by socioeconomic and political considerations. These include, for example, free access to genetic testing, patents for genes and the financial monopoly over molecular medication. Society must propose rules for the potential integration of the knowledge of molecular obstetrics and gynecology into the daily care of those seeking aid or advice. PMID- 12779005 TI - Methods for screening tumors for p53 status and therapeutic exploitation. AB - Mutations in the p53 oncosuppressor gene occur in most human cancers and regulation of the protein is defective in a variety of others. Novel strategies are emerging for the treatment of tumors that have p53 mutations. In this context, the analysis of p53 status is useful in diagnosis and prognosis, and could serve to evaluate the effectiveness of a cancer treatment. In this review, we report an overview of major methods for screening tumors for p53 status and the major strategies suggested for restoring p53 function. PMID- 12779006 TI - Biomarkers in melanoma: staging, prognosis and detection of early metastases. AB - Currently, melanoma remains a surgical disease since early detection and excision of thin melanomas offers the best chance of a cure. Despite intensive clinical investigation, no effective systemic therapies exist for metastatic melanoma. Sentinel lymph node biopsy has greatly aided the staging and prognostic evaluation of primary cutaneous melanoma, however, approximately a third of patients diagnosed with metastatic melanomas present without prior regional lymph node involvement. Additional prognostic biomarkers exist which help determine the risk of advanced melanoma but the accuracy for each current marker is less than 100%. A greater understanding of the biology of melanomas and the development of new methods to identify patients with early (subclinical) metastatic disease may allow for selective and more effective therapy for patients at-risk for advanced disease. In this paper, current and novel potentially more accurate biomarkers for the staging and prognostic evaluation of melanoma patients, and for the detection of subclinical metastases are reviewed. PMID- 12779008 TI - Molecular genetic events in adult acute lymphoblastic leukemia. AB - Treatment of acute lymphoblastic leukemia in adults focuses on the initial assessment of prognostic relevant genetic features as well as response-guided therapy based on molecular data. In at least half of adult acute lymphoblastic leukemia patients, clonal chromosomal abnormalities can be identified that deregulate candidate oncogenes or transcription factors by introducing a heterologous promoter or enhancer. Altered cell cycle progression or upregulated tyrosine kinase activity are other important mechanisms. Most of the translocations can lead to the generation of fusion genes that are translated into chimeric oncogeneic proteins, such as BCR-ABL, providing targets for novel therapeutic agents. PMID- 12779007 TI - Possible role of matriptase in the diagnosis of ovarian cancer. AB - There are increasing data that suggest a role for the serine protease matriptase and its inhibitor, hepatocyte growth factor activator inhibitor-1, in the pathogenesis and progression of ovarian cancer. This review will discuss the matriptase/inhibitor system in the context of ovarian cancer and examine the possibility that this system might be a useful therapeutic and/or diagnostic target in this disease. PMID- 12779009 TI - Molecular detection of West Nile virus RNA. AB - West Nile virus is a mosquito-borne flavivirus that is primarily maintained in nature in a mosquito-bird-mosquito transmission cycle. Mammals, including humans and horses, are incidentally infected through biting by mosquitoes infected with West Nile virus. Since 1994, West Nile virus outbreaks have occurred with a high incidence of severe disease in humans and horses. In the USA, West Nile virus was first detected in 1999 in New York City and has since spread to 39 states in humans. The virus has resulted in over 4161 known human cases and at least 277 human deaths. Surveillance techniques employing nucleic acid-based assays have played an essential role in monitoring the spread of West Nile virus and are displacing the former gold standard cell culture-based assays. In this article we review the current techniques for diagnosis of West Nile virus, focusing on RNA detection, and suggest a number of new directions for genetic diagnosis of West Nile virus. PMID- 12779010 TI - Genetic variation in immune function and susceptibility to human filariasis. AB - The generation of a draft sequence of a the human genome has provided the opportunity to characterize human diversity, even as it pertains to differences in host response to parasitic infection with organisms that cause lymphatic filariasis, malaria and schistosomiasis. Worldwide, human infection with filarial pathogens represents a significant cause of morbidity throughout the tropics. In particular, epidemiologic evidence suggests that a genetic component contributes to susceptibility and possibly the outcomes of filarial infection. Different approaches can be applied in population-based studies in areas where filarial infection is endemic, such as genome linkage scans and candidate gene analysis for the purpose of identifying genetic risk factors. This review summarizes recent advances in our understanding of genetic contributions to human lymphatic filariasis and addresses the immediate questions facing the field. It is anticipated that the identification of susceptibility genes in filarial infection could provide new insights into therapeutic strategies, including pharmacological intervention and vaccine development, and influence public health measures to control or avert infection. PMID- 12779011 TI - Integrating microarrays into disease-gene identification strategies. AB - Positional cloning represents one of the most successful paradigm shifts in identifying the underlying patho-mechanisms in human disease. While traditional discovery tools focused on identifying defects at the tissue or cellular level, positional cloning identifies the damaged region of the genome as the preliminary step. While a large number of inherited single gene disorders have been mapped using this approach, a bottleneck still exists in combing through the genomic interval, often millions of nucleotides in length, to identify the nucleotide changes which result in a defective protein and subsequent disease. Along with the recent unravelling of the human genetic code, the development of massively parallel tools, such as microarrays, represent an equally important step forward in unraveling pathogenic genome dysfunctions. There are many emerging variants on microarray technology, such as expression arrays, exon arrays, array-based comparative genomic hybridization and sequencing arrays. Several of these platforms, if used properly, can accelerate the positional cloning process. The proper use of the platform is driven by knowledge of the underlying molecular defect being searched for and the operating characteristics of the array. The resultant insight forms the basis for improved molecular diagnostics and novel therapeutic targets. PMID- 12779012 TI - DNA diagnosis by capillary electrophoresis and microfabricated electrophoretic devices. AB - DNA diagnosis is experiencing an impressive progression towards the development of novel technology to identify various clinically relevant categories of genetic changes and to meet the exponential growth of genomics. The introduction of capillary electrophoresis has dramatically accelerated the completion of the first draft of the human DNA sequence in the Human Genome Project, and thus, has become the method of choice for analysis of various genetic variants. The recent development of microfabricated electrophoretic devices has led to the possibility of integrating multiple sample handling with the actual measurement steps required for automation of molecular diagnostics. This review highlights the most recent progress in capillary electrophoresis and electrophoretic microdevices for DNA-based diagnostics, including the important areas of genotyping for point mutation, single nucleotide polymorphisms, short tandem repeats and organism identification. The application of these techniques for infectious and genetic disease diagnosis, as well as forensic identification purpose, are covered. The promising development and the challenges for techinical problems are also discussed. PMID- 12779014 TI - Pretreatment with interleukin-2 modulates perioperative immunodysfunction in patients with renal cell carcinoma. AB - Complex perioperative immunodysfunction occurs in patients with renal cell carcinoma undergoing nephrectomy. Here, the effect of pretreatment with IL-2 is addressed. Of 63 patients who underwent tumour nephrectomy, 26 patients received four doses of 10 Mio IE/m2 IL-2 b.d. s.c. (i.e. a total of 40 Mio IE/m2) a week before operation, 37 did not. Parameters of cellular and humoral immunity (differential blood count, T-cell markers CD2, CD3, CD4, and CD8, B-cell markers CD19 and CD20, monocyte markers CD13 and CD14, NK-cell marker CD16, activation markers CD25, CD26, CD69 and HLA-DR, and cytokines IL-1-receptor antagonist (IL 1RA), IL-2, soluble IL-2-receptor (sIL-2R), IL-6, IL-10, and TGFbeta) were measured in peripheral venous blood. Blood was drawn before IL-2, one day before and immediately after the operation, and on the 1st, 3rd, 5th, and 10th postoperative day. All patients showed postoperatively elevated leukocyte and granulocyte counts, and elevated serum levels of cytokines IL-6 and IL-10. T-cell and activation markers were decreased. However, all these alterations were less accentuated in patients who had been pretreated with IL-2. Monocyte counts and IL 2 and TGFbeta levels were decreased, but IL-1RA and sIL-2R levels were elevated in pretreated patients. IL-2-related toxicity was WHO grade I-II in all patients, grade III in one patient. The anaesthetic regimen had no measurable effect. IL-6 concentrations were higher in renal venous than in venous pool blood, indicating IL-6 production in the tumour in vivo. Tumour-specific survival was better in pretreated patients with tumours extending beyond the kidney. Pretreatment with IL-2 modulates perioperative immunodysfunction in patients undergoing tumour nephrectomy. This affects in particular T-cell-mediated immunity and levels of cytokines IL-10 and IL-6. The IL-2 application scheme used here was followed by distinct counter regulation including monocytes, IL-2, sIL-2R, IL-1RA and TGFbeta. Taken together, pretreatment with IL-2 may complement surgery in the treatment of patients with renal cell carcinoma, and may help close the therapeutic gap between neo-adjuvant and adjuvant immunotherapy. PMID- 12779013 TI - The 3' untranslated region of the chicken c-src protooncogene modulates gene expression. AB - Tight regulation of the Src tyrosine kinase activity is essential for a variety of cellular processes, namely transitions of the cell cycle. The peaks of Src activity are dependent on its posttranslational modifications as well as on the regulation of gene expression. The 3'UTRs of mRNAs are often crucial for rapid changes of the protein level. The chicken c-src 3'UTR effects on gene expression have been explored. The c-src 3'UTR decreased the in vivo tumorigenic potential of the src-activated mutants in chickens. This corresponds with the finding that the c-src 3'UTR reduced the Src protein and src mRNA levels and luciferase activity in vitro. Our results suggest that the chicken c-src 3'UTR plays a role in the negative control of gene expression, either transcriptionally or posttranscriptionally. PMID- 12779015 TI - Adjuvant autologous tumour cell-lysate vaccine versus no adjuvant treatment in patients with M0 renal cell carcinoma after radical nephrectomy: 3-year interim analysis of a German multicentre phase-III trial. AB - Even M0 RCC is associated with tumour progression in approximately 30% of all patients after radical nephrectomy. Nevertheless, no effective adjuvant treatment after radical nephrectomy has been established. In a multicentre phase-III trial we investigated the impact of an adjuvant autologous tumour cell-lysate vaccination on the progression-free survival of patients with M0 RCC after radical nephrectomy. Between January 1997 and August 1998 a total of 558 patients with a renal tumour were enrolled at 55 different centres (study group) in Germany. Prior to radical nephrectomy all patients were centrally randomized (Quintiles Germany) to either receive an adjuvant autologous tumour cell-lysate vaccine (6 applications at 4-week intervals after radical nephrectomy) or to receive no adjuvant treatment (control group) after radical nephrectomy. All patients were evaluated following standardized diagnostic investigations at 6 month intervals. Following the inclusion criteria (RCC stages pT2-3bpN0-3M0, TNM classification, UICC 1993), 365 patients were evaluable for the 3-year progression-free survival analysis. There were 240 patients with stage pT2pN0M0 (104 in the vaccine group and 136 patients in the control group) and 89 patients with stage pT3pN0M0 (46 in the vaccine group and 43 patients in the control group). The remaining 36 patients had positive lymph nodes. The trial was performed according to ICH-GCP guidelines. The 3-year progression-free survival rate for all tumour stages was 84.7% in the vaccine group and 80.9% in the control group. Patients with RCC stage pT3pN0-3M0 in the vaccine group demonstrated an advantage (74.4% in the vaccine group vs 65.9% in the control group). For RCC stage pT2pN0-3M0 the 3-year progression-free survival rate in the vaccine group was 89.7% compared to 85.7% in the control group. Follow-up of all patients enrolled in this trial is ongoing. This is the first randomized trial indicating a benefit from an adjuvant vaccination in patients with M0 RCC after radical nephrectomy. The advantage in terms of progression-free survival was more pronounced in patients with T3-tumours. However, it must be emphasized that the results of the final study report (2003) must be awaited before definite recommendations can be made. PMID- 12779016 TI - A prospective open-label single-arm phase II study of chimeric monoclonal antibody cG250 in advanced renal cell carcinoma patients. AB - cG250 is an IgG1 kappa light-chain chimeric monoclonal antibody that binds to a cell surface antigen found on 95% of clear-cell renal cancer. A multicentre phase II study was performed to evaluate the safety and efficacy of repeated doses of cG250. Thirty-six patients with metastatic RCC were included. All patients were nephrectomized for the primary tumour. Twenty-one patients were pretreated (e.g. with IL-2, IFN-alpha). A weekly dose of 50 mg cG250 was given by i.v. infusion for 12 weeks. Patients with SD or tumour response (PR, CR) after 12 weeks of treatment could receive additional treatment for 8 more weeks. None of the 36 enrolled patients had any cG250 grade III or IV toxicity. Only three patients had grade II toxicity possibly related to the study medication. ELISA testing gave no evidence for relevant amounts of HACA. Eleven patients presented with SD and ten were eligible for extension treatment. After the end of the study in the follow up period, one patient demonstrated a CR in week 38 and another patient with SD showed a significant reduction of the overall tumour load in week 44. Six additional patients with progressive disease at study entry were stable for more than six months after the treatment start. The weekly schedule of i.v. cG250 in patients with metastatic RCC was safe, very well tolerated and non-immunogenic in a 12-week treatment regimen. cG250 showed anti-tumour activity. PMID- 12779017 TI - Assessment of nephrotoxicity in the chick embryo: effects of cisplatin and 1,2 dibromoethane. AB - Morphological symptoms of mesonephric kidney damage were analysed in chick embryos treated with nephrotoxic agents--CDDP or DBE. The drugs were administered intraamniotically on ED 3 at doses 0.03 and 0.3 microg CDDP or 100 and 300 microg DBE per embryo. Body weight and absolute and relative measures of the mesonephroi (length, weight and form) were evaluated on ED 10. The higher doses of both agents affected the mass of this organ significantly. Simultaneously, a dose dependent increase of renal malformations was detected in treated embryos, while the incidence of gross and cardiovascular defects was low (DBE) or absent (CDDP). Together with less pronounced effects on the total body growth, the results gave evidence for a higher sensitivity of the mesonephros to toxic insult when compared to the whole organism. A direct cytotoxic effect multiplied by concomitant injury of blood supply seemed to be the main cause of CDDP nephrotoxicity. In the case of DBE, damage to the mesonephros was probably associated with a primary impairment of the vascular network. The chick embryo in ovo provides a promising system for the assessment of nephrotoxic effects induced by prospective therapeutic agents and environmental contaminants during the prenatal period. PMID- 12779018 TI - Morphological, genetic and functional variability of a T-cell hybridoma line. AB - The variability in the morphology, modal number of chromosomes, TCR expression and functional reactivity of a CII-specific T-cell hybridoma at continuous subcultivation have been investigated. As the number of passages increased, besides the oval semiadherent cells (normal phenotype), fibroblast-like cells (transformed phenotype) were also observed. The two cell subpopulations differed in their karyotype characteristic, as well as in their functional reactivity. The cell population with a normal phenotype was characterized by a tetramodal number of chromosomes (30, 40, 48 and 70) and trisomies of chromosomes 6 and 14, while the cell population with a transformed phenotype was characterized by a trimodal number of chromosomes (11, 68 and 74) and trisomy of chromosome 12. A nullisomy of sex chromosomes was established in both types of cells. In the initial passages of subcultivation, 73.04% of the cells with a normal morphological phenotype expressed TCR-CD3 complexes on their surface and possessed high functional reactivity. After a two-week subcultivation, the values of these indices went down considerably: 46.11% of the cells expressed functional TCR-CD3 complexes, as a result of which their functional reactivity decreased. Only 2.71% of the cells with a transformed morphological phenotype expressed functional TCR CD3 complexes on their surface. In these cells, a total loss of reactivity towards the specific antigens was established. The achieved results show that at continuous subcultivation the T-cell hybridomas are unstable, and with the increase in the number of passages there appear chromosome rearrangements, leading to loss of their functional reactivity. PMID- 12779019 TI - Helicobacter pylori: what test for diagnosis and what treatment for eradication? PMID- 12779020 TI - Diagnosis and eradication of Helicobacter pylori in patients with duodenal ulceration in the community. AB - OBJECTIVE: To determine the value of Helicobacter pylori (Hp) serology in diagnosis of active Hp infection in patients with documented duodenal ulcer (DU) and to directly compare the efficacy and side-effects profiles of metronidazole or tinidazole in a triple therapy regimen to eradicate active Hp infection. DESIGN OF STUDY: Prospective, single-blinded, randomised trial. METHODS: One hundred patients from General Practice with documented DU and Hp seropositivity had a C14 Urea Breath Test (UBT). Those who tested positive were randomised to receive one-week, twice daily omeprazole 20 mgs and clarithromycin 250 mgs in combination with metronidazole 400 mgs (OCM) or tinidazole 500 mgs (OCT). Eradication was confirmed by a repeat UBT. RESULTS: Eighty five sero-positive patients had a positive pre-treatment UBT. On intention to treat basis, OCT (100%) had a significantly better eradication rate than OCM (87.8%), p = 0.023. There was no difference in side effects. CONCLUSION: (1) Positive Hp serology in patients with DU does not always mean active infection and (2) for patients in the community with active Hp and DU disease OCT is significantly better than OCM for eradicating Hp. PMID- 12779021 TI - Breast diseases: a histopathological analysis of 3279 cases at a tertiary care center in Pakistan. AB - OBJECTIVE: To know the frequency of breast diseases in Pakistani females. METHODS: A retrospective analysis of 3279 breast specimens received over a period of 4 years (1993-1996) at the department of pathology, the Aga Khan University Hospital. RESULTS: Out of a total of 3279 breast specimens, common breast lesions included infiltrating duct carcinoma 37%, followed by fibro adenoma 16.95%, fibrocystic change 13.96%, mastitis 6.83% and duct ectasia 5.33%. Majority of the cases of infiltrating duct carcinoma were encountered in the 5th and 6th decades of life. Tumour size was 2 or >2 cms. in 93% of cases and 40% of them showed 3 or >3 positive lymph nodes. Grade I tumours were 11.38%, grade II 59.17% and grade III tumours 29.47%. Correlation of grade with lymph node metastases (3 or >3+ve nodes) showed 15 cases (1.53%) of grade I, 178 cases (18.25%) of grade II and 68 (6.97%) cases of grade III tumours. CONCLUSION: This study shows that in Pakistani females, the most commonly encountered lesion in carcinoma of the breast followed by the benign lesions such as fibro adenoma, fibrocystic disease & others. Breast carcinoma occurs at a younger age group with predominance of high-grade lesions and with frequent lymph node metastasis. PMID- 12779022 TI - National Polio Day campaign in a squatter settlement through medical students. AB - OBJECTIVE: To provide maximum polio vaccination coverage to under-five (<5) year children of a squatter settlement through Ziauddin Medical University (ZMU) medical students. SETTING: ZMU has established a Primary Health Care Program in a squatter settlement, which is predominantly inhabited by migrants from North Western Province of Pakistan and Afghanistan. The total population is approximately 20,000 and the proportion of <5 year children is nearly 19%. METHODOLOGY: ZMU started Oral Polio Vaccination (OPV) campaigns from 1996; up to 1999. The OPV campaigns were confined to the National Immunization Days (NIDs). A different strategy of "door-to-door" OPV services by medical students was undertaken in 1997. In December 1999 the polio vaccination coverage of <5-year children was evaluated through duster sampling. This paper describes the steps for improving OPV coverage in Sikanderabad. RESULTS: The estimated number of <5 year children in the area is 4,600. In a duster sample survey after the 1999 NID campaign out of 620 under 5 year children living in 429 households, 529 (85%) received OPV, with a 95% CI for OPV coverage of 82 to 88%. The coverage before the campaigns initiated by ZMU was 52%. CONCLUSION: The results indicate that the polio coverage of <5-year children has risen with the change in strategy. Door-to door coverage strategy with the help of medical students proved effective in providing vaccination. No case of suspected poliomyelitis has been reported from the area since October 1999. PMID- 12779023 TI - Serum aminotransferase levels and platelet count as predictive factor of fibrosis and cirrhosis in patients with chronic hepatitis C infection. AB - OBJECTIVE: To assess the ratio of aminotransferases in combination with platelets count as a predictive factor for stages of fibrosis in patients with chronic hepatitis C virus infection. METHODS: A total of 266 patients were evaluated retrospectively who had undergone liver biopsy as a part of their evaluation for the management of chronic hepatitis C infection at Shifa International Hospital from 1998 to 2002. All these patients had complete blood counts and liver function tests including transaminases. The ratio of AST/ALT and platelet count of <150,000 were noted and degree of the fibrosis and cirrhosis on liver biopsies were matched. None of these patients had history of alcohol consumption. The fibrosis stages used were according to METAVIR score. RESULTS: An AST/ALT ratio of >1 and a platelet count of <150,000 had sensitivity of 85.6 and specificity of 90.0 with positive predictive value of 91.2 and negative predictive value of 83.4 for the fibrosis grade-III and IV. An AST/ALT ratio of <1 and a platelet count of >150,000 for grades 0-2, the sensitivity was 87.3, specificity 60.0, PPV 87.3 and NPV 48.0. CONCLUSION: An AST/ALT ratio of >1 in combination with a platelet count of <150,000 can predict advance stage of fibrosis and cirrhosis in patients with chronic hepatitis C infection. In these patients, a liver biopsy may not be necessary. PMID- 12779024 TI - Optimization of low pre-operative hemoglobin reduces transfusion requirement in patients undergoing transurethral resection of prostate. AB - OBJECTIVE: To identify factors that influence peri-operative hemorrhage in view of reducing the need for transfusions in patients undergoing trans uretheral resection of prostate (TURP). METHODS: All patients undergoing TURP between January 1997 and December 1999 were identified using ICD 9CM coding and indexing system. Overall 430 patients were identified, however, 384 charts were included and reviewed for demographics, pre and intra-operative data and post-operative morbidity. Patients were divided into two groups on the basis of presence of significant hemorrhage. RESULTS: Overall 384 patients were analyzed. Nineteen patients had hemorrhage--group I whereas 365 had no significant hemorrhage--group II. Mean age and co-morbidities in the two groups were similar. However, in group I, 58% presented with urinary retention compared to 33% in group II. In group I, factors that reached statistical significance include; operative time (p<0.05), mean resected tissue weight (p<0.02), and patient presentation (urinary retention) (p<0.032). There was no significant difference in the two groups with respect to type of anesthesia (regional versus general) and histology of the resected tissue. Patients with mean pre-operative hemoglobin of 10.6 % had a 37% transfusion rate. CONCLUSION: Operative time, weight of resected prostate tissue are inter related and are only partly controllable. Low pre-operative hemoglobin is the only reversible factor in reducing transfusion following TURP. PMID- 12779025 TI - Injection practices and sharp waste disposal by general practitioners of Murree, Pakistan. AB - OBJECTIVE: To describe the injection and sharp waste disposal practices of general practitioners of Murree, Pakistan. METHODOLOGY: In-depth interviews of all general practitioners available between 10th to 20th July 2000 were conducted. Practitioners were interviewed about injection administration and disposal of waste due to injections and other sharp material. RESULTS: Twenty general practitioners out of 25 were interviewed. All claimed using disposable syringes only once. None of them was disposing off syringes in sharp containers in the clinic. Of 20, 12 (60%) were throwing syringes at open places and 5 (25%) in municipal waste bins. Most of the injections by general practitioners were administered for fever, body aches, diarrhea and respiratory tract infection. CONCLUSION: Sharp waste disposal is not safe in Murree. Improper disposal of sharp waste needs development of cost effective methods that are applicable at a small scale. Larger studies are required to quantify the gravity of the problem. PMID- 12779026 TI - Sonographic prevalence of acquired cystic renal disease in patients receiving haemodialysis. AB - OBJECTIVE: To determine the sonographic prevalence of Acquired Cystic Renal Disease (ACRD) in patients on maintenance haemodialysis and its relationship to the duration on dialysis. METHODS: All patients with end stage renal disease (ESRD) who were receiving maintenance haemodialysis in Urology Department of Mayo Hospital Lahore between November 1997 to February 1998 were screened by ultrasound for the prevalence of ACRD. RESULTS: Forty patients with ESRD on maintenance haemodialysis were evaluated. The mean age of patients was 39.9 years. The male to female ratio was 2.3:1. Four patients (10%) had documented ACRD. The proportion of patients with ACRD increased with increasing duration of dialysis; ACRD was found in 60% of patients who had dialysis for more than 3 years, 20% in those who had been dialyzed for 1-3 years, and no patient developed ACRD who was on dialysis for less than one year. All patients remained asymptomatic except one, who had intermittent macroscopic haematuria. There was no evidence of neoplasm in any of these patients on clinical grounds or on ultrasound. CONCLUSION: We found a low prevalence of ACRD in the population studied compared to previously published series. This can be explained by the fact that most of our patients were on dialysis for less than one year. A further study is suggested with greater numbers in a center where patient stay on dialysis for longer period and patient turnover is low. PMID- 12779027 TI - Male breast cancer. AB - OBJECTIVE: To review cases of male breast cancer. SETTINGS: Histopathology Section, Department of Pathology, The Aga Khan University, Karachi. METHOD: Fifty one cases of male breast cancer specimens, received during a period of 10 years, routinely processed and stained with Haematoxylin and Eosin were analyzed. Special stains and Immunohistochemistry were used in difficult cases. RESULTS: Male breast cancer affected individuals in the sixth and seventh decades of life with a mean age of 56.2 years. Infiltrating ductal carcinoma (IDC) was the predominant type. Skin involvement was seen in 27.45% of the cases. Breast lump was the most common presenting symptom followed by skin ulceration. At the time of presentation 43.13% patients had a tumour size of more than 3 cm. CONCLUSION: Male breast cancer is a rare disease. Most of our findings correspond to the published local and international data. PMID- 12779028 TI - Chromosomal abnormalities as a cause of recurrent abortions: a hospital experience. AB - OBJECTIVE: To analyze the chromosomal abnormalities in couples with history of recurrent abortions. DESIGN: Retrospective study. SETTING: Cytogenetic section, Department of Pathology at the Aga Khan University Hospital, Karachi, Pakistan. PATIENTS: Three hundred couples with history of recurrent abortions. MAIN OUTCOME MEASURE: Structural chromosomal abnormalities in carrier couples. RESULTS: G banded chromosomes were analyzed from both partners of 300 couples who had had two or more spontaneous abortions. Sixteen (5.3%) couples showed a chromosomal aberration in one of the partners. Seven translocations, 5 inversions, 2 deletions and 2 sex chromosomal abnormalities were identified. There was an increase in structural aberations, which was not significantly different from that reported worldwide. CONCLUSION: Chromosomal analysis is advocated as a primary tool in the evaluation of couples with repetitive fetal wastage. PMID- 12779029 TI - Acute angioedema in paraphenylenediamine poisoning. PMID- 12779030 TI - Implications for the practice of a patient expectation and satisfaction survey, at a teaching hospital in Karachi, Pakistan. AB - OBJECTIVE: To study the expectations and satisfaction of patients visiting Family Practice Clinics, at the Aga Khan University Hospital, in Karachi, Pakistan. METHODOLOGY: A cross sectional survey of 316 patients was carried out. A questionnaire, based on the study objectives was developed and administered. The participating patient signed a consent form, after assurance of confidentiality was provided. Data on the demographic profile of the patients was collected. RESULTS: The mean age was 33.81 years, with 105 (33.2%) women, and 211 (66.8%) men. The majority were married, with education above intermediate level and were in private or government service or were housewives. The median for the patient waiting time was 30 minutes, against an expectation of 12.69 minutes. Reading newspaper, watching television, reading magazine, reading Quran and listening to music were quoted as ways to lessen the burden of waiting to see a physician. The average consultation time with the physician was 13.89 minutes, against an expectation of 16.37 minutes. Patient expectation in terms of listening by the doctor with patience, explanation of the diagnosis and treatment, prescription of medicines, ordering of investigations and specialist referral has been documented. Objections to the presence of medical student, nursing student, resident doctor, nurse and an observer, in the consultation room have also been documented. Reasons quoted for the objection include issues of privacy/confidentiality, lack of justification, discomfort, and interference with the consultation process. The expected average cost for doctor's consultation was Pakistani Rs. 124, while 196 (61.8%) of the respondents were satisfied with the consultation based on Rs. 70. CONCLUSION: We have collected important information to improve the services offered at our Family Practice clinics. PMID- 12779031 TI - Seasonal variation in bacterial pathogens isolated from stool samples in Karachi, Pakistan. AB - OBJECTIVE: To determine the seasonal variation of the commonly isolated bacterial pathogens in stool samples. MATERIAL AND METHODS: A retrospective descriptive study was undertaken of all the stool samples submitted from within Karachi to the Aga Khan University Hospital Laboratory over a period of five years (January 1997- December 2001) in order to determine the commonly isolated bacterial pathogens and to predict their seasonal variation. RESULTS: A total of 16379 stool samples were included in this review. Bacterial isolates were found in 6670 stool samples (culture detection rate=40.7%). The mean age at the time of culture of each sub-group was < or = 1 year group (6.58 +/- 3.1 months), 1-5 years (2.13 +/- 0.94 years), 5-14 years (8.3 +/- 2.6 yrs) and adults (43.2 +/- 18.5 years). Male: Female ratio was 1.2:1. Vibrio cholera 01 Ogawa (32.8%), Campylobacter jejuni (17.3%), Enteropathogenic Escherichia coli (9.9%), Salmonella paratyphi b (6.6%) and Shigella flexneri (6.2%) were the most common organisms isolated. These organisms show a distinct seasonal variation with summer predilection. CONCLUSION: In contrast to the previous studies from South Asia, which have identified E. coil, followed by Vibrio cholerae as the most common enteric isolates, we found Vibrio cholera 01 Ogawa followed by Campylobacter jejuni as the most common enteric pathogens isolated in an urban setting. It is important to consider seasonal variation when empirically treating diarrheal diseases in our region. PMID- 12779032 TI - Anesthesia--past, present and future. AB - In 1680 Sydenham wrote, "Among the remedies which has pleased almighty God to give to man to relieve his sufferings, none is so universal and so efficacious as opium." This appraisal is still true today. Morphine, the principal analgesic agent of the opium alkaloids, has provided the structural model from which modern synthetic opiates have been designed and synthesized. Morphine, as well as the synthetic opioids, has become a permanent part of the armamentarium of an anesthesiologist. The inhalational anesthetics (nitrous oxide and ether) were not introduced until the mid 1800s, and the cardiovascular stabilizing qualities of ether have led to the use of ether as the structural model for synthesizing many of the inhalational agents used today. Aside from the important discoveries of intravenous and inhalational agents, the introduction of capnograph and pulse oximeter in the 1980s has significantly improved morbidity and mortality in anesthesia. With the improved anesthetic management, surgeries have been pushed to new heights, which would have never been possible without creative and optimal anesthetic management. Unfortunately, surgery and anesthesia are continued to be pressured by medical economics to cut costs, while there is increased risk of managing the critically ill patients. This trend will continue; therefore, anesthesiologists today must rise to the challenge. PMID- 12779033 TI - Endovascular treatment of the carotid artery rupture with massive hemorrhage. AB - BACKGROUND: To assess the efficacy and safety of endovascular embolization in the treatment of the carotid artery rupture. METHODS: A series of 21 patients who had carotid artery rupture (CAR) with massive transoronasal or transcervical hemorrhage, all treated with endovascular embolization, were reviewed. There were 17 males and 4 females ranging in age from 21 to 79 years, with a mean of 47 years. The predisposing factors of the CAR were head and neck tumors associated with surgical treatment and/or post-radiation therapy (n = 17), surgical complications (n = 1) and trauma (n = 3). Detachable balloons with liquid adhesives were used in 13 patients for vascular occlusion and obliteration of pseudoaneurysm of the internal or common carotid artery. Liquid adhesives and/or coils were used in 8 patients, in whom only the branches of the external carotid artery were involved (n = 6), or obliteration of a pseudoaneurysm with preservation of the ICA (n = 2). RESULTS: Endovascular treatment was technically successful in all 21 cases of the CAR with cessation of profuse hemorrhage immediately after embolization. No recurrence of bleeding was observed. Two patients developed permanent hemiparesis several hours later after occlusion of the carotid artery (10%). Three patients had a transient ischemic attack; all returned to normal neurological conditions later (14%). Ten patients having advanced head and neck carcinoma died during follow-up period owing to the disease itself. Clinical follow-up period for these patients lasted 2 to 48 months, with a mean of 16 months. CONCLUSIONS: Therapeutic endovascular embolization of the CAR is the primary invasive modality for treating CAR with profuse hemorrhage. It was proven both safe and effective over a relatively long follow-up period. PMID- 12779034 TI - Prognostic factors for Vogt-Koyanagi-Harada disease. AB - BACKGROUND: Vogt-Koyanagi-Harada disease is a chronic bilateral granulomatous panuveitis with involvement of the central nervous system, auditory and integumentary systems. The prognosis depends mostly on the development of complications or not. The purpose of this study was to identify the prognostic factors of Vogt-Koyanagi-Harada (VKH) disease. METHODS: Retrospective chart review of all the patients diagnosed with VKH disease between 1991 and 2000 was performed. Data recorded included age, sex, clinical features, systemic manifestations, recurrence, HLA typing, treatment, complications, and final visual acuity. Statistical analysis was performed using Statistical Products and Services Solutions. RESULTS: There were thirty-nine patients, the mean age at presentation was 39.82 +/- 12.38 years. The majority (46.2%) of patients were probable VKH disease, followed by incomplete type (43.5%), with only 10.3% complete type. Ten patients had at least one complication, including cataract in 19 eyes, glaucoma in 4 eyes and choroidal neovascularization in 3 eyes. Fifty eyes (64.1%) had final visual acuity of 6/12 or better. The interval between ocular symptom and treatment had significant effect on the development of complicaion (p = 0.022) and recurrence (p = 0.003), which were also significantly associated with the final visual acuity (p = 0.007). For patients who were diagnosed and treated within two weeks, the extent of exudative retinal detachment correlated significantly with final visual acuity (p = 0.001). CONCLUSIONS: The visual prognosis of VKH disease is generally favorable. The development of complications or recurrence was significantly associated with the visual outcome. Interval between development of ocular symptom and treatment seemed to be an important predictor in the early stage of VKH. It is important to start the treatment early even when the typical features of VKH disease have not completely appeared. PMID- 12779035 TI - Glycemic control with different premixed insulin in Taiwanese people with type two diabetes mellitus. AB - BACKGROUND: The aim of this study is to compare the clinical responses between Mixtard 50 HM (premixed insulin with 50% regular insulin (RI) and 50% neutral protamine Hagedom (NPH)) and Mixtard 30 HM (premixed insulin with 30% RI and 70% NPH) among type 2 diabetic patients. The acceptability of NovoPen 3 and traditional syringe was also evaluated among these patients. METHODS: Twenty-six patients were injected with Mixtard 30 HM initially using traditional syringe, which was switched to Mixtard 50 HM eight weeks later. When the switch commenced, patients were randomly assigned to two groups. One group used traditional syringe and the other used NovoPen 3. Blood glucoses were measured before breakfast, before lunch and at bedtime for two days in a week by patients themselves. Hemoglobin A(1c)(HbA(1c)) was checked at week 8 and week 16. At the end of the trial, patients completed 1 questionnaires regarding their acceptability of NovoPen 3. RESULTS: There was no statistical difference in mean blood glucose levels measured before breakfast, before lunch, or at bedtime, or in HbA(1c), between Mixtard 30 HM and Mixtard 50 HM treatment periods. However, blood glucose after breakfast declined more in Mixtard 50 HM treatment period than in Mixtard 30 HM treatment period. Up to 96.2% of the study subjects reported that NovoPen 3 was more convenient in use than traditional syringe. CONCLUSIONS: Blood glucose declined more from pre-breakfast level to pre-lunch level when patients used Mixtard 50 HM instead of Mixtard 30 HM. We suggest that Mixtard 50 HM is more appropriate for post-breakfast hyperglycemic diabetic patients. NovoPen 3 results in a similar clinical glycemic response but is better accepted by the patients than conventional syringe. PMID- 12779036 TI - A radiological study after anterior cruciate ligament reconstruction. AB - BACKGROUND: Disruption of the anterior cruciate ligament is a common injury, often resulting in functional instability. Currently, arthroscopic anterior cruciate ligament (ACL) reconstruction using the central third bone-patellar tendon-bone graft is a common surgical procedure. The purpose of this study was to evaluate the postoperative structural remodeling in the ligament following ACL reconstruction over time by X-ray and magnetic resonance imaging (MRI). METHODS: We performed arthroscopic ACL reconstruction with central third bone-patellar tendon-bone autograft for 27 patients (14 males, 13 females, mean 27.2 years old) between October 1995 and January 1999. All patients were examined post operatively at 3, 6, 12, and 18 months by X-ray to evaluate the sequential change of the tibia bone tunnel and by MRI to monitor the sequential signal changes in the recipient and donor sites. RESULTS: The MRI showed an increased signal at the patellar tendon (donor site) at 3 and 6 months and decreased signal after 1 year. The signal of ACL graft from MRI remained increased after operation. On AP, lateral view of X-ray, the diameter at the joint site of the tunnel was larger than at the bone plug site. Pearson correlation method revealed that the diameter of the bone tunnel expanding at the joint site was statistically greater than that at the bone plug site (p < 0.05). The difference between the bone tunnel diameter at the bone plug site and the joint site was not statistically correlated with the distance between them (by linear regression method, p = 0.77). CONCLUSIONS: We concluded that the windshield wiper effect could not be the only factor inducing the enlargement of the bone tunnel, based on our results. The signal change of patellar tendon was more obvious than that of the bone-patellar tendon-bone graft. PMID- 12779037 TI - Metallic stenting for treatment of central venous obstruction in hemodialysis patients. AB - BACKGROUND: Central venous obstruction is a serious and common complication in hemodialysis patients. The recurrence rate is high after balloon dilation, while surgical repair is hazardous in these chronically ill patients. The aim of this study was to evaluate the efficacy of primary Wallstent placement for treatment of hemodialysis-related central venous obstructions. METHODS: Eighteen hemodialysis patients with symptomatic shunt dysfunction and arm swelling due to subclavian (n = 3) or innominate (n = 15) venous obstructions were treated at Kaohsiung Veterans General Hospital with primary Wallstent placement from November 1998 to August 2001. Technical success and complications were reported. Primary and secondary patency rates of stent and hemodialysis access were calculated by survival analysis with Kaplan-Meier method. RESULTS: A total of twenty-five Wallstents were deployed for central venous stenosis (n = 18) and occlusion (n = 7) in these 18 hemodialysis patients. The initial technical success rate was 100%. Fourteen episodes of re-obstruction developed during the observation period. Of them, seven episodes occurred within the stent, four episodes outside of but abutting to the peripheral end of the stent, and another three episodes of re-obstruction were attributed to delayed shortening of the Wallstent. Six of the 14 episodes were treated percutaneously with angioplasty alone, while seven episodes necessitated additional stent placement. No stent migration or other complications were encountered. Primary patency rates of stent and hemodialysis access at 3, 6, 12 and 18 months were 100 and 89%, 73 and 68%, 49 and 42%, and 16 and 0%, respectively. Secondary patency rates of stent and hemodialysis access each were both 100% after 3 months, 93 and 100% after 6 months, 85 and 91% after 12 months and, 68 and 72% after 24 months. CONCLUSIONS: Wallstents are safe to deploy, with excellent technical success for hemodialysis related central venous obstructions. The Wallstent provides continued use of a hemodialysis access for a substantial period. However, repeated interventions may be necessary to maintain the patency. PMID- 12779038 TI - Mitral valve repair--early and medium-term results. AB - BACKGROUND: Mitral valve repair is the treatment of choice in patients with mitral insufficiency. Mechanical valves, even when functioning perfectly, are less efficient hemodynamically than a native mitral valve and also have attendant morbidity from thrombolism, anticoagulant-related hemorrhage, and endocarditis. Long-term data have confirmed the durability of mitral repair. METHODS: Mitral valve repair was performed on 89 consecutive patients (20 patients in ischemic group) between Nov. 1997 and Jan. 2002, In all patients, posterior annuloplasty with Capentier ring (30) or polytetrafluoroethylene Teflon strip (59) was performed. The repair technique included quadrangular resection, artificial chordae replacement, chordal transfer, and chordal shortening. RESULTS: There were 5 early deaths (4 deaths in the ischemic group), 9 major complications (5 complications in the ischemic group). Mean follow-up in the entire patient series was 25.18 months. There was one late death and one thromboembolic cerebral episode. Two patients required reoperation after mitral repair. There was a significant improvement in New York Heart Association functional class and grade of mitral regurgitation. CONCLUSIONS: The results of our study confirm that mitral valve repair results in acceptable operative mortality rate, excellent early and medium-term results and incidence of post operative complications. PMID- 12779039 TI - Aggressive multiple myeloma with brain involvement in a young patient. AB - Multiple myeloma is generally a neoplastic plasma cell disorder of the old age. It seldom attacked young people or manifested intracranial plasmacytomas. Here we report a 28-year-old young patient of plasma cell dyscrasia who presented multiple plasmacytomas of bones while sparing bone marrow. Cerebral involvement with intratumorous hemorrhage complicated the clinical course. The unique clinical features are presented and literature is reviewed. PMID- 12779040 TI - Entrapped vaginal pessary presented with frequency and urge incontinence. AB - Pessaries remain helpful for patients with genital prolapse who refuse or are not able to have surgery due to medical complications. It is generally considered both conservative and safe to wear a vaginal pessary. Only a few cases of severe complications have been reported whereby most result from a neglected pessary. We present a patient with extreme frequency and urge incontinence with recovery following the removal of the entrapped pessary. An 82-year-old oriental female carrying a vaginal pessary for more than 10 years had suffered from intractable frequency and urge incontinence for 6 months. The urinary analysis and urodynamic studies showed no sign of lower urinary infection or outlet obstruction. Physical examination found a black ring pessary in the vagina which could not be mobilized due to its deep embedment. The lower urinary tract symptoms were recovered after surgical removal of the pessary. Complication of a vaginal pessary may be caused by an ignorant patient or doctor. Frequency and urge incontinence could be an early sign of a severe complication due to a neglected pessary. PMID- 12779041 TI - An unusual buccal adenoma with extensive squamous metaplasia and cyst formation. AB - Salivary gland tumors are the second most common neoplasm of the mouth following squamous cell carcinoma, and account for a significant proportion of oral tumors. We report a case of an unusual adenoma that presented as a solitary intraoral buccal mass in a 28-year-old male patient. The tumor was intrepreted as an unusual pleomorphic adenoma because of the absence of connective tissue changes and presence of metaplastic epithelial change which is an occasional finding of such tumors. Moreover, the presence of cystic structures with papillary infoldings filled with some crystals and pearl-like keratinized material were also salient features. PMID- 12779042 TI - Nasopharyngeal lymphoid hyperplasia of an HIV carrier, mimicking nasopharyngeal cancer. AB - Lymphoid hyperplasia of cervical lymph nodes is a common head and neck presentation of human immunodeficiency virus (HIV) infection. Lymphoid hyperplasia of HIV carrier presenting as nasopharyngeal tumor and neck mass is rarely discussed. We reported a 30-year-old male patient who had left side neck mass for 3 months. Nasopharyngeal cancer with neck metastasis was impressed due to huge nasopharyngeal tumor and neck mass. The patient was then transferred to our clinic for further evaluation. Physical examination revealed a 30 x 30 mm mass over left jugulodigastric area and a 20 x 20 mm mass over left posterior triangle of neck. Telescopic examination revealed a 20 x 20 mm bulging mass over nasopharynx. Nasopharyngeal biopsy was done and histopathologic examination revealed lymphoid hyperplasia. Laboratory survey showed seropositive of HIV infection for both enzyme-linked immunosorbent assay (ELISA) and western blot tests. PMID- 12779043 TI - Synthesis and nicotinic binding studies on enantiopure pinnamine variants with an 8-azabicyclo[3.2.1]octane moiety. AB - Bioisosteric replacement of the 9-azabicyclo[4.2.1]nonane pharmacophoric element of the novel alkaloidal marine toxine pinnamine (5) by the 8 azabicyclo[3.2.1]octane moiety resulted in conformationally restricted analogues 6a and 6c of (-)-ferruginine (4). Key step in the diastereoselective synthesis of these pyranotropanes was the condensation of enantiopure ecgonine methyl ester (9) from the "chiral pool" with the lithium anion of N-tert-butylbutyraldimin and subsequent cyclisation with TFA. The potential nAChR ligands were tested for their in vitro affinity for the (alpha4)2(beta2)3 and the alpha7* nAChR subtypes. Despite obvious structural similarities with the potent alkaloids pinnamine (5) and (-)-ferruginine (4) the pyranotropanes 6a and 6c exhibited distinctly lower nAChR affinities. PMID- 12779044 TI - Synthesis and analgesic activity of stereoisomers of cis-fluoro-ohmefentanyl. AB - Four stereoisomers 1a-d of cis-fluoro-ohmefentanyl have been synthesized. Their absolute configurations were determined by X-ray analysis of (3S,4R,2'S)-(-)-cis I d. The analgesic activity (mice, sc, hot plate) revealed extreme stereodifferences. The ED50 value of (3R,4S,2'S)-(+)-cis-I (1a) was 0.000774 mg/kg (17958 times more potent than that of morphine), while the corresponding antipode 1c was almost inactive. PMID- 12779045 TI - Synthesis, characterization and mutagenicity of new cis-[Pt(2-substituted benzimidazole)2Cl2] complexes. AB - In this study, four new platinum(II) complexes with the structures cis [Pt(Ligand)2Cl2] (ligand = 2-(p-methoxy-/or-p-chlorobenzyl or p methoxyphenyl)benzimidazol (1, 2, 4 respectively) and 5(6)-methyl-2 phenoxymethylbenzimidazole (3) were synthesized and characterized by their elemental analysis, and IR and 1H NMR spectra. The potentials of the Pt(II) complexes for short-term bacterial mutagenicity were tested in reverse-mutation assays using Salmonella typhimurium frame-shift strain T 98 and S. typhimurium TA 100 and TA 102 strains, which carry mutations particularly sensitive to reversion by DNA base-pair substitution. The tests were performed in the absence of S9 rat liver fraction. Among the complexes tested 1 had no mutagenic activity. Complex 4 was found to be weakly mutagenic in TA 98 only. The Pt(II) complexes 2 and 3 were found to be mutagenic in TA 98, TA 100 and TA 102. PMID- 12779046 TI - Photochemistry and in vitro phototoxicity studies of levomepromazine (methotrimeprazine), a phototoxic neuroleptic drug. AB - The neuroleptic drug levomepromazine (1, previously known as methotrimeprazine) is photolabile under UV-A and UV-B light in aerobic conditions. Irradiation of a methanol solution of this drug produces one photoproduct, resulting from the oxidation of 1 to its sulfoxide parent. It is demonstrated that photodegradation occurs via type II mechanism involving irreversible trapping of self photogenerated singlet molecular oxygen. 1 shows a photohemolytic effect on human erythrocytes and photoinducers lipid peroxidation. PMID- 12779047 TI - Physico-chemical properties and spectrophotometric determination of biologically active 1-alkyl-2-(2-pyridyl)pyridinium bromides. AB - Physico-chemical properties of compounds prepared from 2,2'-bipyridine, 1-alkyl-2 (2-pyridyl)pyridinium bromides, were investigated by DC polarography and by GC MS. Their ionization potentials were calculated. Additionally, the formation of associates with bromothymol blue and methyl orange during the spectrophotometric determination was measured. It was determined that 1-alkyl-2-(2 pyridyl)pyridinium ions are reduced by 2 one-electron steps in a DC polarography system. The reduction potentials are not related to the ionization potential values calculated for the substances investigated. The carcinogenic potential (tg alpha) of the parent compound 2,2'-bipyridine and of a series of 1-alkyl derivatives was very low indicating that the compounds are not carcinogenic. The MS fragmentation patterns indicate the low stability of the 1-alkyl substituents. It was shown that 2,2'-bipyridine is either fragmented to two pyridine ions or the--N=CH--fragments are removed. Additionally, spectrophotometric determinations of colored associates of 1-alkyl-2-(2-pyridyl)pyridinium bromides with bromothymol blue and methyl orange were investigated and the optimal condiditions for these determinations are reported. PMID- 12779049 TI - Effect of vehicles and penetration enhancers on the in vitro percutaneous absorption of celecoxib through human skin. AB - The aim of this study was the comparison of three different formulations (gel, o/w emulsion, oleagenous cream) and two penetration enhancers (oleic acid and menthol) as vehicle systems for celecoxib in respect of release and penetration through excised human skin in vitro. The influence of the vehicle on the release rate was studied in vitro using a cellulose acetate membrane. The release rate could be increased by up to 6.5 and 2.5 times with gel and o/w emulsion compared to oleagenous cream respectively. Further in vitro penetration measurements using human skin on Franz diffusion cells were performed with and without oleic acid and menthol as enhancers. It was shown that the penetration rate is strongly dependent upon the enhancer type and concentration but not on the vehicle itself and could be increased by 48% when 5% oleic acid was used in oleagenous cream. In all formulations tested, celecoxib was released and penetrated into human skin more quickly and to a greater extent from the gel formulations. There is no topical formulation available of celecoxib and its penetration properties through human skin have not been investigated. Since celecoxib creates some gastrointestinal disturbances, topical formulations of celecoxib preferably in gel form including 5% oleic acid could be suggested as an alternative. PMID- 12779050 TI - Studies on gynaecological hydrophilic lactic acid preparations, part 6: use of Eudragit E-100 as lactic acid carrier in intravaginal tablets. AB - Hydrophilic intravaginal tablets based on methylcellulose and containing lactic acid component with Eudragit E-100 undergo swelling in standard conditions. A high flow-limit of the gel that originates from the tablets as well as its dynamic viscosity should allow for the durable dosage form in the vagina. By choosing a 1:1 ratio of lactic acid to Eudragit E-100, it is possible to obtain tablets disintegrating into a gelform at physiological range of 3.8-4.4. An increase in the amount of lactic acid in the complex in relation to the polymer up to 2:1 and 3:1 ratios results in gels with a lower pH. These gels possess an acid reserve that might be ableto neutralise the excess of alkali present in severe vaginal infections. PMID- 12779048 TI - Hydrophilic gel containing nanocapsules of diclofenac: development, stability study and physico-chemical characterization. AB - The purpose of this work was to develop and to characterize hydrophilic gels containing nanocapsules (NC) of diclofenac (DIC). Nanocapsules suspension of poly epsilon-caprolactone containing free acid diclofenac were prepared by nanoprecipitation. The pH value of the nanocapsules suspension was 5.70 +/- 0.03 and the mean sizes of the NC were in the sub 300 nm range. Drug incorporated into the nanocapsules was close to 100% and the encapsulation efficiency was 104.1% +/ 3.5%. Diclofenac nanocapsules suspension (1 mg/mL) was incorporated in a Carbopol gel matrix fournishing a formulation with 0.5 mg of DIC/g. The gel stability was evaluated in terms of the macroscopic and microscopic aspect, rheological properties, pH and drug recoveries. As a result, we obtained a suitable formulation for topical use presenting a non-Newtonian behaviour with plastic properties and with intact nanostructures in the gel matrix after 3 months storage atroom temperature (freeze-fracture electron microscopy). PMID- 12779051 TI - Structure activity relationship studies of cinnamic acid derivatives as inhibitors of human neutrophil elastase revealed by ligand docking calculations. AB - Structure-activity relationship of cinnamic acid derivatives as inhibitors of the human neutrophil elastase is reported. Comparison of the inhibitory concentrations (IC50 values) with the results of the ligand docking calculations revealed that the structure element of the aromatic ortho-dihydroxy groups combined with a lipophilic residue seems to be a prerequisite for an optimal binding within the active site. PMID- 12779052 TI - Detrimental effects of water extracts from surface and interior of Taxus baccata leaves on the two-spotted spider mite (Tetranychus urticae Koch). AB - Surface deposits on Taxus baccata needles removed by dipping in water of 96, 60 or 40 degrees C for 5 s caused changes in life history components of mites. Paclitaxel was among other peaks present in the removed fractions in concentrations between 0.017 and 0.170 microg/g of fresh weight (f.w.) increasing with temperature. Long extraction for 60 min at only 40 degrees C did not increase removable paclitaxel, but at 60 degrees C extraction rate was the highest (1.326 microg/g) suggesting that leakage from an interior of needles occurred. Mortality, developmental time, total fecundity, oviposition period and life history parameters of Tetranychus urticae Koch. were detrimentally affected. PMID- 12779053 TI - Antitrypanosomal naphthylisoquinoline alkaloids and related compounds. AB - In view of the need to develop new drugs against human African trypanosomiasis, a series of naturally occurring naphthylisoquinoline alkaloids, axially chiral acetogenic products derived from tropical plants, have been investigated for their activity against Trypanosoma brucei brucei TC 221. Likewise compounds corresponding to the two molecular portions, the naphthalene and the isoquinoline parts were tested, as well as molecules related to the central biaryl core of the alkaloids. Among all compounds tested, the natural, genuine alkaloids themselves, in particular dioncophylline B with its biaryl system and a moderate number of free hydroxy functions, showed the highest activities. Our results demonstrate that naphthylisoquinoline alkaloids constitute an interesting novel class of antitrypanosomal compounds worth further optimization. PMID- 12779054 TI - Antituberculosis agents IV: in vitro antimycobacterial activity and cytotoxicity of N-piperazinyl quinolone derivatives containing 2-thienyl and 2-furyl moiety. AB - A series of N-[2-(2-furyl)-2-oxoethyl], N-[2-(2-furyl)-2-oxyiminoethyl], N-[2-oxo 2-(2-thienyl)ethyl] and N-[2-oxyimino-2-(2-thienyl)ethyl] piperazinyl quinolones (1a-h; 2a-h) were evaluated for antituberculosis activity against M. tuberculosis H37Rv using the BACTEC 460 radiometric system and BACTEC 12B medium. Our results indicated that compounds 1a, 1e and 1g were efficient antimycobacterial agents showing MIC values ranging from 0.78 to 6.25 microg/ml. In general, ciprofloxacin derivatives were more active than norfloxacin derivatives and the oxime analogues were less active than corresponding ketones. Active compounds (1a, 1e and 1g) were also screened by serial dilution to assess toxicity to VERO cell line. The cytotoxicity of tested compounds indicated that compound 1a was the less toxic compound (IC50 > 62.5 microg/ml). This compound was tested for efficacy in vitro in TB-infected macrophage model (EC90 = 3.25 microg/ml). PMID- 12779055 TI - Sesquiterpenes from roots of Lingularia veitchiana. AB - Together with seven known sesquiterpenes, a new guaiane, a new furanoeremophilane, and a new eudesmane were isolated from the roots of Ligularia veitchiana. Their structures were elucidated by spectroscopic methods. The bioactivities of three known guaiane sesquiterpenes were determined. PMID- 12779056 TI - Improved synthesis and in vitro evaluation of quinuclidin-2-ene based ligands for the nicotinic acetylcholine receptor. PMID- 12779057 TI - Influence of drug solubility on the release of slightly water-soluble drugs from HPMC matrices. PMID- 12779058 TI - Enalapril maleate form II: stabilization in a tablet formulation. PMID- 12779059 TI - Sublingual delivery of 17beta-estradiol from cyclodextrin containing tablets. PMID- 12779060 TI - Hypericum perforatum L. and Chamomilla recutita (L.) Rausch.--accumulators of some toxic metals. PMID- 12779061 TI - Inhibition of UV-induced mutagenesis in Euglena gracilis by benzothiazole derivatives. PMID- 12779062 TI - A new sterol from the pseudobulb of Desmotrichum fimbriatum Blume. PMID- 12779063 TI - Megastigmane and flavone glycosides from Acanthus ilicifolius. PMID- 12779064 TI - The problem of diagnosing chronic pancreatitis. AB - High alcohol consumption in Europe makes it necessary to consider chronic pancreatitis when a patient with history of alcohol abuse presents with abdominal pain. Diagnosis should be made on the basis of imaging procedures and function tests. A scoring system for this, together with a short evaluation of the present diagnostic procedures, are presented. PMID- 12779065 TI - Flat colorectal neoplasia: identification, pathogenesis and clinical significance. PMID- 12779066 TI - Evaluation of dyspeptic symptoms in patients with and without Helicobacter pylori infection and normal upper gastrointestinal endoscopy. AB - BACKGROUND: A relationship between Helicobacter pylori infection and dyspeptic symptoms has not yet been demonstrated. AIM: To evaluate any possible difference in symptom score between dyspeptic patients with and without H. pylori infection who have normal upper gastrointestinal endoscopy and no other appreciable gastrointestinal or systemic disease. PATIENTS: A series of consecutive patients affected by upper abdominal disturbances completed a symptoms questionnaire before undergoing upper gastrointestinal endoscopy with a rapid urease test to detect H. pylori infection. Patients with normal upper gastrointestinal endoscopy and abdominal ultrasound were included in the study. The symptoms assessed were burping and belching, bloating, odynophagia, dysphagia, postprandial fullness, heartburn, early satiety, nausea, vomiting, regurgitation, sour taste in mouth, epigastric pain at fasting, epigastric pain postprandial, epigastric pain nocturnal, and pain in right hypocondrium and were scored in terms of intensity and frequency on a scale from 0 to 4. RESULTS: The total number of patients who met the inclusion criteria was 263 out of 1187 examined. A total of 113 H. pylori positive and 150 H. pylori-negative patients were compared. Among the symptoms evaluated, belching and bloating and heartburn were present in more than 50% of patients of both groups. No statistical difference was found in terms of presence or absence of each symptom, and intensity or frequency between H. pylori-positive and -negative patients. CONCLUSION: H. pylori infection does not seem to be associated with a specific symptom in patients with upper abdominal complaints and normal upper gastrointestinal endoscopy. PMID- 12779067 TI - Oesophageal acid-peptic strictures in the histamine H2 receptor antagonist and proton pump inhibitor era. AB - BACKGROUND AND AIM: We present a survey on the incidence, demography and natural history (re-dilatation rates) of patients with oesophageal acid-peptic stricture seen between 1977 and 1995. PATIENTS AND METHODS: Prospective, cohort observational study. Stricture severity was graded mild, moderate or severe (needing forcible dilatation with bougies). Most were treated with histamine H2 receptor antagonists or proton pump inhibitors, and were followed up by serial check endoscopy. RESULTS: A total of 156 of 7429 (2%) reflux patients had stricture: mild, n=56; moderate, n=25; severe, n=75. The prevalence of reflux disease is rising, paralleled by an increase in the number of patients with stricture; hence the incidence of stricture is unchanged: 1977-1982, 1.8% (18/986); 1983-1989, 2.4% (61/2595); 1990-1995, 2% (77/3848). Demography: mean age 68 years; mean length of reflux history 6.4 years; 15% on non-steroidal anti inflammatory drugs/aspirin; 18% with Barrett's metaplasia. Mean follow-up: 5.6 years. OUTCOME: of the 75 with severe stricture, 36 (48%) needed forcible dilatation only once, and 39 more often (13 twice, nine three times), most within 2 years. Only six of these patients need > or = 6 re-dilatations. CONCLUSION: The incidence of stricture is low and remains unchanged, despite the increased prevalence of reflux disease. Most patients have a good prognosis, only half needing further forcible dilatation, mainly within the first 2 years. PMID- 12779068 TI - Acid exposure and altered acid clearance in GERD patients treated for Helicobacter pylori infection. AB - BACKGROUND: After the eradication of Helicobacter pylori, an increased incidence of gastroesophageal reflux disease and acid gastric secretion have been reported. AIM: To evaluate the effect of Helicobacter pylori-eradication on proximal and distal gastroesophageal reflux and acid clearance in patients with gastroesophageal reflux disease. PATIENTS AND METHODS: Sixty-eight gastroesophageal reflux disease patients (age range 18-61 years) were studied by upper endoscopy. All underwent esophageal manometry and dual probe 24-h pH-metry. RESULTS: Percent of time at pH<4 was significantly increased in the proximal esophagus of Helicobacter pylori-eradicated patients compared to Helicobacter pylori-negative (2.4+/-0.5 vs. 1.0+/-0.2; p<0.01); no differences were found in the distal esophagus (14.0+/-3.7 vs. 9.0+/-1.4%, NS). The total number of reflux episodes was significantly higher in the proximal oesophagus of Helicobacter pylori-eradicated patients (37+/-3 vs. 22+/-3, p<0.05). In the distal esophagus, acid clearance was significantly longer, both during total time (1.4+/-0.2 vs. 0.8+/-0.7 min, p<0.01), and in the supine period (8.5+/-2.7 vs. 2.7+/-0.4 min, p<0.05). No differences were reported in the manometric parameters of the two groups of patients. CONCLUSION: In patients with gastroesophageal reflux disease, Helicobacter pylori eradication is associated with increased acid exposure of the proximal esophagus and delayed distal acid clearance. PMID- 12779069 TI - Dyspepsia and Helicobacter pylori infection: a prospective multicentre observational study. AB - OBJECTIVES: Dyspepsia still represents an unsolved clinical enigma. AIM: The aims of this study were to determine whether symptoms and Helicobacter pylori infection are predictors of organic disease in uninvestigated dyspepsia, and if H. pylori eradication improves symptoms in functional dyspepsia. METHODS: An observational study was performed on outpatients with uninvestigated dyspepsia. Symptoms were scored and H. pylori status determined. Patients with functional dyspepsia and H. pylori infection were randomly given either a standard eradicating treatment or a 1-month course of empirical treatment. The latter was also given to functional dyspeptic patients without infection. Symptoms were re assessed in functional dyspeptic patients at 2- and 6-month follow-up visits. Patients receiving eradicating treatment were re-tested for H. pylori at the 2 month visit. RESULTS: A total of 860 patients were studied and 605 (70.3%) were affected by functional dyspepsia. H. pylori infection was diagnosed in 71.8% of patients with organic dyspepsia and in 65.0% with functional dyspepsia (p=0.053). Male sex, anaemia, smoking habit, age over 45 years, and severe epigastric pain, but not H. pylori infection, were independent predictors of organic disease. Symptoms significantly improved in most functional dyspeptic patients regardless of their H. pylori status and type of treatment. CONCLUSION: H. pylori infection is not a strong predictor of organic disease in uninvestigated dyspepsia. H. pylori eradication is not essential to improve symptoms in functional dyspepsia. PMID- 12779070 TI - Colorectal flat neoplasia. AB - BACKGROUND AND AIMS: An attempt has been made to evaluate the clinicopathological characteristics of flat colorectal neoplastic lesions, and analyse the factors associated with the malignancy. PATIENTS AND METHODS: A total of 115 flat neoplastic lesions, > or = 5 mm in size, diagnosed in 87 patients by colonoscopy, were investigated. RESULTS: The rectum was the most common location. Almost half (49.6%) of the flat neoplasms were small (5-10 mm), 27.8% were 11-20 mm and the remainder (22.6%) larger than 20 mm. The surface was smooth in 55.7%, granular in 20.0% and nodular in 24.3%. Histologically, the flat lesions were tubular, tubulovillous and villous adenomas in 69.6%, 20.9% and 5.2%, respectively. Five lesions (4.3%) were composed of carcinomas without adenoma. High-grade dysplasia, intramucosal carcinoma and invasive carcinoma were diagnosed in 9.6%, 7.8% and 6.1% of all flat neoplasms, respectively. Univariate analysis demonstrated that the location, size, surface pattern and histologic type of the flat lesions were factors associated with malignancy. However, in multivariate analysis, the size of the flat lesions was the only significant risk factor for malignant transformation. CONCLUSIONS: Flat neoplastic lesions of the colorectum have a relatively high rate of malignancy, and size is the most important factor associated with malignancy. PMID- 12779071 TI - Increased susceptibility to nonalcoholic fatty liver disease in heterozygotes for the mutation responsible for hereditary hemochromatosis. AB - BACKGROUND: Insulin resistance is a key feature of nonalcoholic fatty liver disease. Patients with hereditary hemochromatosis, a disease characterized by progressive iron overload due, in most cases, to homozygosity for C282Y mutation in the HFE gene, have often decreased insulin sensitivity and release. AIMS: To determine whether increased iron parameters/heterozygosity for the mutations of the HFE gene confer susceptibility to nonalcoholic fatty liver disease. PATIENTS: One hundred and thirty-four consecutive Italian patients with clinical and ultrasonographic diagnosis of nonalcoholic fatty liver disease (82 with hyperferritinemia), half confirmed by liver biopsy. METHODS: Insulin was determined by radioimmunoassay. HFE gene mutations were determined by polymerase chain reaction and restriction fragment length polymorphism analysis. RESULTS: (1) Prevalence of C282Y HFE mutation was significantly higher in patients with nonalcoholic fatty liver disease compared to controls, the difference being more striking in patients with hyperferritinemia than in those without. (2) The presence of mild iron overload was associated with a lower insulin release. (3) Carriers of C282Y mutation developed nonalcoholic fatty liver disease despite lower body mass index and triglycerides. CONCLUSION: The mild iron overload associated with heterozygosity for C282Y HFE mutation confers susceptibility to nonalcoholic fatty liver disease, causing relative insulin deficiency. PMID- 12779072 TI - Natural history of pancreatitis associated with cystic fibrosis gene mutations. AB - BACKGROUND: An increased incidence of CFTR mutations has recently been reported in chronic and idiopathic pancreatitis. AIM: The aim of the study was to verify these data and describe the clinical, morphological and histological findings in 99 patients (59 males, 40 females, mean age 40+/-16 years), 45 suffering from idiopathic chronic pancreatitis and 54 from acute recurrent pancreatitis. METHODS: Each subject was screened for the 18 CFTR mutations: DF508, DI507, R1162X, 2183AA>G, 21303K, 3849+10KbC>T, G542X, 1717-1G>A, R553X, Q552X, G85E, 711+5G>A, 3132delTG, 2789+5G>A, W1282X, R117H, R347P, R352Q), which cover 72% of cystic fibrosis chromosomes in the Italian population, plus the 5-thymidine allele in intron 8 of the CFTR gene (IVS85T). RESULTS: Among the 99 patients, we found 14 patients with CFTR mutation (14.1%). Three idiopathic chronic pancreatitis patients had cystic fibrosis (compound mutations in two and a single mutation with a pathological sweat test in one) and 11 (11.1%) presented a single mutation (carriers) (seven idiopathic chronic pancreatitis and four acute recurrent pancreatitis). The incidence of patients with cystic fibrosis was 167.5 times higher than that observed in the general population, whereas the carrier frequency was 4.43 times higher for chronic pancreatitis and 2.11 times for acute recurrent pancreatitis than that observed in 428 unrelated partners of cystic fibrosis patients. The prevalence of IVS8-5T was similar (7.1%) to that of the general population (10%). All idiopathic chronic pancreatitis patients with one or more CFTR gene mutations had a long history of recurrent attacks of pancreatitis. The length of recurrences of pancreatitis before diagnosis of chronic pancreatitis was shorter in chronic pancreatitis patients with one or more CFTR gene mutations than in the other idiopathic chronic pancreatitis patients (7.4+/-5.8 vs. 2.1+/-2 years). In idiopathic chronic pancreatitis patients with one or more CFTR gene mutations, exocrine and endocrine insufficiency (diabetes and steatorrhoea) were rare or delayed events. CONCLUSIONS: The natural history of pancreatitis associated with CFTR gene mutations seems to be characterised by recurrences of pancreatitis which develops into chronic pancreatitis. PMID- 12779073 TI - Prospective evaluation of magnetic resonance cholangiopancreatography in patients with biliary disease: comparative study with conventional ultrasonography and endoscopic retrograde cholangiopancreatography diagnostic algorithm. AB - BACKGROUND: Magnetic resonance cholangiopancreatography is an accurate technique that can replace invasive diagnostic methods of the biliary and pancreatic duct. AIMS: Our aim was to assess sensitivity and specificity of magnetic resonance cholangiopancreatography and ultrasonography using the results of endoscopic retrograde cholangiopancreatography as reference, and to establish a diagnostic algorithm under which circumstances magnetic resonance cholangiopancreatography can replace endoscopic retrograde cholangiopancreatography. PATIENTS: Eighty three patients with suspicion of biliary disease based on clinical, biochemical and ultrasonography findings were studied. METHODS: Ultrasonography, magnetic resonance cholangiopancreatography and endoscopic retrograde cholangiopancreatography were performed, comparing the results of the techniques for the determination of their sensitivity and specificity. RESULTS: Sensitivity and specificity results obtained by magnetic resonance cholangiopancreatography were: 100 and 92.8% when dilated ducts were detected (n=61); 97.4 and 97.2% in the diagnosis of choledocholithiasis (n=38); 100 and 96.7% in malignant lesions (n=14) and 81.8 and 98.4% when biliary ducts were normal. The percentage of images of diagnostic quality was 97.6%. Sensitivity and specificity achieved by ultrasonography was: 100 and 57.1% in detection of dilatation, 71 and 97.2% in choledocholithiasis, 92.8 and 96.7% in malignancy and 66.6 and 96.8% in normal ducts. CONCLUSIONS: Magnetic resonance cholangiopancreatography is a technique with high sensitivity and specificity in the evaluation of biliary ducts. Thus, magnetic resonance cholangiopancreatography may replace diagnostic endoscopic retrograde cholangiopancreatography for purely diagnostic purposes, following an initial clinical and ultrasonographic exam. PMID- 12779075 TI - Helicobacter pylori infection in patients with liver cirrhosis: facts and fictions. AB - Helicobacter pylori infection could play a role in different clinical alterations observed in cirrhosis, from gastroduodenal lesions to hepatic encephalopathy. Although its prevalence in cirrhotics is similar to that in controls, H. pylori infection is responsible for the increased prevalence of peptic ulcer observed in these patients. The ammonia production by H. pylori urease does not seem to increase blood ammonia levels during cirrhosis, indicating that its role in hepatic encephalopathy could be marginalized in clinical practice. Dual and triple therapies have been shown to be equally effective for H. pylori eradication in these patients. PMID- 12779074 TI - Lower intestinal bleeding due to aorto-enteric fistula. AB - The case is described of a man who complained of intermittent fever and fatigue. After three digestive endoscopies and computed tomography, a 99m technetium-HM PAO-labelled white cell scan was usefully employed to establish diagnosis. Anaerobic aortic Graft infection and anaemia due to lower intermittent occult intestinal bleeding were found. The intestinal bleeding was caused by secondary aorto-jejunal fistula. This condition is rare, but should be suspected whenever a patient with aortic prosthesis presents with occult digestive bleeding and unexplained fever. PMID- 12779076 TI - A novel approach of targeted ablation of mammary carcinoma cells through luteinizing hormone receptors using Hecate-CGbeta conjugate. AB - Recent studies have shown that human and animal mammary gland carcinoma cell line express luteinizing hormone receptors (LHRs). We have examined the cytotoxic effect of Hecate-CGbeta conjugate, that is, fusion of a lytic peptide (Hecate) and a 15-amino acid fragment of the CGbeta-chain in vitro. To test the hypothesis that the Hecate-CGbeta conjugate selectively abolishes cells possessing LHR, estrogen dependent and independent human breast cancer cell lines (MCF-7; MDA-MB 231) and a mouse Leydig tumor cell line (BLT-1) were treated in vitro with Hecate CGbeta conjugate and Hecate alone. Cytotoxic effects of the Hecate-CGbeta conjugate and the Hecate alone was measured by lactate dehydrogenase (LDH) release immediately after treatment. We observed that the Hecate-CGbeta conjugate selectively, in dose-dependent manner destroys cells possessing LHR in lower concentrations of preparate comparing to the Hecate alone and that the cytotoxic effect is strongly correlated with the number of LHR. Using Western blot analysis we characterized the LHR on membranes of MDA-MB-231, MCF-7 and BLT-1 tumor cell lines. In addition, we showed the evaluation of inhibition potential of the Hecate-CGbeta conjugate to LHR. At a concentration of 33 microM the conjugate inhibited (50%; IC50) the binding of CG to LHR. We suggest further development of this novel approach for the treatment of breast cancer by the Hecate-CGbeta for in vivo trials. PMID- 12779078 TI - Enterolactone in breast cyst fluid: correlation with EGF and breast cancer risk. AB - The purpose of our study was to investigate whether enterolactone does accumulate into breast cyst fluid and whether it correlates with breast cancer risk. We included 258 women who had at least one cyst aspiration and known intracystic cation and epidermal growth factor (EGF) concentration values. For 191 of such women serum aliquots were also available. The median value of serum enterolactone was 17 nM/l (range 1-140 nM/l). The median intracystic level of enterolactone was much higher (63 nM/l, range 0-872 nM/l) and was significantly higher in type I cysts (p = 0.000). This cyst type contained also significantly higher levels of EGF (p = 0.000). A direct relationship was found between serum and cyst fluid enterolactone levels (p = 0.000) and between cyst enterolactone and EGF levels (p = 0.03), the latter correlation being evident especially in type II cysts. Twelve patients in the cohort of women were found to have developed a breast cancer. After univariate analysis breast cancer risk was associated with cyst type and especially with EGF concentration. No association was evident for enterolactone concentration. However, enterolactone concentration appeared to significantly decrease the risk of patients with high EGF concentrations. Our results show that enterolactone does accumulate in breast cysts, and that it modulates the risk related to the intracystic level of EGF, which is confirmed to be a strong predictor of breast cancer risk. PMID- 12779077 TI - Sequential changes in serum triglyceride levels during adjuvant tamoxifen therapy in breast cancer patients and the effect of dose reduction. AB - Tamoxifen (TAM) and estrogen increase serum triglyceride (TG) levels, sometimes inducing severe hypertriglyceridemia and possibly contributing to death. Decreasing the dose of estrogen in estrogen replacement therapy may minimize its adverse effects, including the hypertriglyceridemia. The serum TG, total cholesterol, low-density-lipoprotein (LDL) cholesterol and high-density lipoprotein (HDL) cholesterol were measured periodically in 116 breast cancer patients on adjuvant TAM therapy at a dose of 10 mg twice daily. The serum TG level was significantly increased 15 months after treatment, but the magnitude of increase was clinically insignificant in most patients (n = 102). If TG levels rose above 400 mg/dl, the dose of TAM was reduced to 10 mg once daily (n =14). The TG was lowered to a safer level after the dose reduction of TAM in 10 of 14 patients. The other four patients had early onset hypertriglyceridemia after beginning TAM (within 6 months), and their TG levels did not decrease satisfactorily after dose reduction. Hence, they required antilipemic medication with or without discontinuing TAM. This study suggests that reducing TAM from 10 mg twice daily to 10 mg once daily decreases the marked hypertriglyceridemia that occurs in some patients during TAM treatment. PMID- 12779079 TI - Differential effects of raloxifene, tamoxifen and fulvestrant on a murine mammary carcinoma. AB - The purpose of this study was to evaluate the effect of the selective estrogen receptor modulators raloxifene and tamoxifen and of the pure antiestrogen fulvestrant on tumor growth and progesterone receptor (PR) expression in an experimental model of breast cancer. The effects of these compounds on cell proliferation were studied in primary cultures of a progestin-dependent mammary carcinoma tumor line, in the presence of medroxyprogesterone acetate (MPA) or 17 beta-estradiol (E2). In in vivo studies the tumor was inoculated subcutaneously in BALB/c female mice treated with 20 mg MPA depot. Raloxifene (12.5 mg/kg) or tamoxifen (5 mg/kg) were administered in daily doses or E2 silastic pellets (5 mg) were implanted. When the tumors reached about 25-50 mm2 MPA was removed in half of the animals. E2 induced complete tumor regressions, tamoxifen inhibited tumor growth in vivo while raloxifene disclosed proliferative effects in animals in which MPA had been removed. In vitro, E2 inhibited cell proliferation at concentrations higher than 10(-14)M. Raloxifene and fulvestrant, but not tamoxifen, partially reverted E2-induced inhibition. Fulvestrant and tamoxifen inhibited MPA-induced cell proliferation while raloxifene had a stimulatory effect. Tamoxifen and E2 increased, raloxifene induced no effect, and fulvestrant significantly decreased PR expression. In this study we provide evidence for differential effects of tamoxifen and raloxifene on experimental mammary tumors. Since raloxifene is under evaluation for use in breast cancer prevention, these results may have important clinical implications. PMID- 12779080 TI - Comparison of p53 mutational status with mRNA and protein expression in a panel of 24 human breast carcinoma cell lines. AB - We analyzed the p53 mutational status, mRNA and protein expression in 24 human breast carcinoma cell lines. Following measurement of their DNA content with flow cytometry, we ascertained the copy numbers of the centromere of chromosome 17 (cen17) and p53 with fluorescence in situ hybridization (FISH). A functional yeast assay (FASAY) was used to screen for inactivating mutations. Positive results were subsequently verified by DNA sequencing. Finally, we assessed the mRNA expression with a competitive reverse transcription-polymerase chain reaction (RT-PCR) assay and the protein expression with immunocytochemical staining, western blot, and quantitative flow cytometry. The DNA content of the cell lines ranged from 0.85 to 2.58. Nine cell lines had concordant copy numbers (between two and four) of p53 and cen17, whereas 12 had more, and three less cen17 than p53 copies. The FASAY was successful in all but one cell line and revealed the presence of mutated alleles in 16 of them, 13 cell lines expressed only the mutated, and three both the mutated and the wild-type alleles. The mutations were comprised of 11 missense, two nonsense, and three frameshift mutations. Immunocytochemical staining, western blot and quantitative flow cytometry yielded comparable p53 protein expression results. However, both the mRNA and the protein expression levels varied considerably in the different cell lines and no consistent pattern with regard to the respective p53 mutational status became evident. The results obtained in these breast carcinoma cell lines indicate that no clear-cut linear relationship exists between the p53 mutational status and the extent of its respective mRNA and protein expression. Therefore, direct DNA analyses and functional assays remain the only methods for the reliable detection of p53 mutations. PMID- 12779081 TI - Axilla surgery severely affects quality of life: results of a 5-year prospective study in breast cancer patients. AB - No long-term prospective study has investigated arm morbidity and patient quality of life. It is unclear to what extent breast cancer patients suffer from arm problems, how long such problems affect their lives, and whether quality of life improves as arm problems abate. This prospective cohort study aims to provide data on the clinical factors associated with arm dysfunction, to estimate its prevalence and to relate arm morbidity to quality of life. The Munich Cancer Registry records clinical details of all cancer patients in and around Munich. Quality of life information was provided directly by breast cancer patients (n = 990) over 5 years. Arm morbidity, including movement limitations, swelling and lymph drainage, and quality of life (EORTC QLQ-C30) were assessed. Up to 5 years after diagnosis, 38% of patients were still experiencing arm problems (swelling and limited movement). Consistently over the 5 years, quality of life was significantly (p < 0.001) lower for patients with arm difficulties. For those whose arm problems dissipated, quality of life significantly improved (p < 0.01). A logistic regression analysis showed that extent of axilla surgery (p < 0.003), comorbidity (CVD and diabetes) (p < 0.003), employment (p < 0.01), younger age (p < 0.02), and operating clinic (p < 0.05) significantly contributed to arm problems. Axilla surgery should be re-evaluated since arm morbidity has such a profound effect on patient quality of life. PMID- 12779082 TI - hOGG1 Ser326Cys polymorphism and breast cancer risk among Asian women. AB - To evaluate the potential association between breast cancer risk and Ser326Cys polymorphism of hOGG1 gene, encoding for an enzyme involved in the base excision repair of 8-hydroxyguanine, hospital based case-control studies were conducted in two Asian populations consisting of 475 breast cancer cases (271 Korean and 204 Japanese) and 500 controls (314 Korean and 186 Japanese). PCR-based methods were employed for the genotyping analyses and the statistical evaluations were performed by unconditional logistic regression model. The frequency of hOGG1 Ser/Ser, Ser/Cys, and Cys/Cys genotypes were 22.5, 48.7, and 28.8% in all cases, and 23.7, 52.1, and 24.1% in the controls. No statistically significant associations between the genotypes and breast cancer risk were observed, neither when the ethnic groups were examined separately nor when the total study population was included. Neither did stratification by menopausal status reveal any association between hOGG1 genotypes and breast cancer. Our novel findings therefore suggest that hOGG1 Ser326Cys polymorphism is unlikely to play a modifying role in individual susceptibility to breast cancer among Asian women. PMID- 12779083 TI - PPARgamma ligands and ATRA inhibit the invasion of human breast cancer cells in vitro. AB - Invasion and metastasis are the main causes of death in breast cancer patients. Increased expression of matrix metalloproteinases (MMPs), especially gelatinases (MMP-2 and -9), has been closely associated with tumor progression. One of the nuclear hormone receptors (NHR), peroxisome proliferator-activated receptor gamma (PPARgamma), is a ligand-activated transcriptional factor that regulates cell proliferation, differentiation and apoptosis in both normal and cancer cells. Recent data indicate that PPARgamma activation by its ligands can also lead to the inhibition of gelatinase B (MMP-9) and the blockage of migration in macrophages and muscle cells, implying the possibility that PPARgamma ligands may possess anti-invasive activities on tumor cells. In this study, we showed that treatment of the highly aggressive human breast cancer cell line MDA-MB-231 with the synthetic PPARgamma ligands pioglitazone (PGZ), rosiglitazone (RGZ), GW7845 or its natural ligand 15-deoxy-delta 12, 14-prostaglandin J2(15d-PGJ2), at concentrations at which no obvious cytotoxicity was observed in vitro, led to a significant inhibition of the invasive capacities of this cell line through a reconstituted basement membrane (Matrigel) in a Transwell chamber model. All trans-retinoic acid (ATRA), a ligand for retinoic acid receptor (RAR), was also studied and showed a similar inhibitory effect on invasion. Although no change was observed in the expression of MMP-9 after challenge with PPARgamma ligands and/or ATRA on this cell line, the natural tissue inhibitor of gelatinases, namely the tissue inhibitor of MMP 1 (TIMP-1) was upregulated by these treatments and the gelatinolytic activities of gelatinases in the conditioned media were decreased. Since MMP-2 was not detectable in the conditioned media of MDA-MB-231 cells, and the gelatinolytic activities of the conditioned media were reduced only by MMP-9 neutralizing antibodies, it is most likely that the reduction of gelatinolytic activities by PPARgamma ligands and/or ATRA was due to the decrease of MMP-9 activities. Because MMP-9 was absolutely required in the transmigration of this cell line through Matrigel in our in vitro model as demonstrated by neutralizing antibodies against MMP-2 and -9, we concluded that down-regulation of gelatinase activities is, at least in part, responsible for the reduction of the invasive capacities of MDA-MB-231 cell line in vitro. Our results, for the first time, indicate that PPARgamma ligands may have therapeutic value for the treatment of highly invasive breast cancer by targeting its invasive behavior. PMID- 12779084 TI - SR protein expression and CD44 splicing pattern in human breast tumours. AB - Altered gene expression during breast tumour progression can occur through alternative splicing of mRNAs. The SR proteins have been identified as important factors in RNA splicing and in the incorporation of alternative exons in experimental systems. We have studied SR protein expression by western blot in human breast cell lines and in a cohort of 101 invasive breast tumours to examine the relationship with alternatively spliced isoforms of the CD44 gene. Multiple SR proteins (SR75, 55, 40, 30) were expressed in most cell lines and tumours, and their relative expression was independent of grade, size, or nodal status. Higher relative expression of SR55 protein was associated with an altered pattern of CD44 variants incorporating exon v7 (p = 0.047) as determined by reverse transcription-polymerase chain reaction (RT-PCR) and Southern blot analysis. Nevertheless, transient transfection of MCF7 and HBL100 breast cell lines with SR55 had no direct effect on the expression of CD44 v7 variant expression. We conclude that while SR proteins may be important and necessary factors in mRNA splicing, other factors are also necessary to influence the regulation of alternatively spliced isoforms of CD44. PMID- 12779085 TI - Expression profiling of breast cancer cells by differential peptide display. AB - Expression profiling of RNAs or proteins has become a promising means to investigate the heterogeneity of histopathologically defined classes of cancer. Peptides, representing degradation as well as processing products of proteins offer an even closer insight into cell physiology. Peptides are related to the turnover of cellular proteins and are capable to reflect disease-related changes in homoeostasis of the human body. Furthermore, peptides derived from tumor cells are potentially useful markers in the early detection of cancer. In this study, we introduced a method called differential peptide display (DPD) for separating, detecting, and identifying native peptides derived from whole cell extracts. This method is a highly standardized procedure, combining the power of reversed-phase chromatography with mass spectrometry. This technology is suitable to analyze cell lines, various tissue types and human body fluids. Peptide-based profiling of normal human mammary epithelial cells (HMEC) and the breast cancer cell line MCF-7 revealed complex peptide patterns comprising of up to 2300 peptides. Most of these peptides were common to both cell lines whereas about 8% differed in their abundance. Several of the differentially expressed peptides were identified as fragments of known proteins such as intermediate filament proteins, thymosins or Cathepsin D. Comparing cell lines with native tumors, overlapping peptide patterns were found between HMEC and a phylloides tumor (CP) on the one hand and MCF-7 cells and tissue from a invasive ductal carcinoma (DC) on the other hand. PMID- 12779086 TI - Transformation of non-cancerous human breast epithelial cell line MCF10A by the tobacco-specific carcinogen NNK. AB - Repeated treatments of non-cancerous human breast epithelial cells MCF10A with a low dose of the tobacco-specific carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1 butanone (NNK) induced the development of cancerous cells. NNK-transformed MCF10A cells acquired cancerous properties including anchorage-independent cell growth and increased cell motility. Cellular transformation of MCF10A cells was accompanied by a loss of responsiveness to 17beta-estradiol and decreased rate of cell proliferation. NNK-transformed MCF10A cells were also tumorigenic in immunodifficient mice. Studies of changes in the regulation of intracellular signaling pathways revealed that the upstream Erk pathway was down-regulated in the NNK-transformed cells. Our data provide the first evidence suggesting that the tobacco carcinogen NNK is competent to induce malignant transformation of non cancerous human breast epithelial cells. Our findings suggest that the tobacco carcinogen NNK may contribute to early events in human breast carcinogenesis. PMID- 12779087 TI - Localization of BRCA1 protein in human breast cancer cells. AB - There is still an ongoing debate concerning the cellular localization of BRCA1 protein in breast cancer. To address this question, we compared the localization of BRCA1 protein using several monoclonal (Ab-1) or polyclonal (C20, D20, I20) antibodies under different technical conditions on human breast cancer cell lines. We worked on the fixation and permeabilization conditions in order to preserve the morphological structures of the cells, as confirmed by transmission electron microscopy studies. As expected from the gene sequence analysis and the biochemical features, both nucleus and cytoplasmic BRCA1 protein staining were detected in cells fixed for 60 min in 4% paraformaldehyde and permeabilized with either 0.3% saponin or 0.02% Triton. In these conditions, the same results were obtained: (i) with the four antibodies tested, (ii) with several dilutions (up to tenfold) of the monoclonal antibody, and (iii) in all the tested breast cancer cell lines. In addition, we validated the functionality of these conditions by quantifying the effects of estrogens and their antagonists on the regulation of BRCA1 protein expression in the MCF7 cell line. PMID- 12779088 TI - Androgen pathway dysregulation in BRCA1-mutated breast tumors. AB - BACKGROUND: Using array analysis for screening RNA from BRCA1-mutated and sporadic breast tumors, we observed that AIGF/FGF-8 expression was lost in BRCA1 mutated breast tumors. Since this growth factor is induced by androgens, we studied the androgen receptor (AR) expression in BRCA-mutated tumors and in matched sporadic breast tumors. METHODS: Paraffin embedded breast tumors of carriers of a BRCA1 mutation (n=41, median age of patients at time of surgery was 41 years [range 28-59 years]) or a BRCA2 mutation (n=14, median age 41 years [range 31-85 years]) were analyzed for the presence of ER-alpha, PR, P53 and AR using standard immunohistochemical techniques. All statistical tests used, Pearson chi2 and Fisher exact, were two-sided. RESULTS: The AR was only present in 12% of BRCA1-mutated tumors, with mutations located at the C-terminal half of the BRCA1-gene. The AR expression was significantly more prevalent, however, in a series of 61 sporadic breast tumors (80%) and in BRCA2-mutated tumors (50%). In contrast to an increased percentage of p53 positive cells, in 66% of the BRCA1 mutated tumors, the ER-alpha expression was observed only in 25% and the PR in 13% of these specimens. The three steroid hormone receptors were expressed in about half of the BRCA2-mutated specimens studied. CONCLUSIONS: Our data add to the emerging evidence that the biological phenotype of BRCA1-associated tumors may be different from BRCA2 and non-hereditary cases. The loss of the AR expression, as shown by immunohistochemistry, together with the observed loss of other steroid hormone receptors in BRCA1-mutated tumors may lead to a hormone independent growth or to anti-hormone resistant growth of these tumors. PMID- 12779089 TI - E1AF expression levels are not associated with prognosis in human breast cancer. AB - E1AF is a transcription factor involved in regulation of several metastasis associated genes, and is associated with overexpression of HER2/neu. We were unable to find a clear prognostic value of E1AF expression in human breast cancer. Furthermore, no association of E1AF levels with HER2/neu mRNA levels, hormone receptor status, histological grade, tumor size, or lymph node involvement was found. PMID- 12779090 TI - Generation of CD8+ cytotoxic T lymphocytes against breast cancer cells by stimulation with mammaglobin-A-pulsed dendritic cells. AB - Mammaglobin-A is exclusively expressed by breast cancer cells. Thus, mammaglobin A-specific T cell immune responses may be useful for the design of new breast cancer-specific immunotherapies. We show herein that CD8+ T cells generated against recombinant mammaglobin-A-pulsed dendritic cells display a marked cytotoxic activity against mammaglobin-A-positive breast cancer cell lines. This study indicates the immunotherapeutic potential of this novel antigen for the treatment of breast cancer. PMID- 12779091 TI - Parathyroid hormone assays--evolution and revolutions in the care of dialysis patients. AB - Renal osteodystrophy may present with low, normal, or high bone turnover. An ideal parathyroid hormone (PTH) assay should discriminate between the bioactive whole PTH-(1-84) molecule and PTH fragments, including the PTH-(7-84) fragment. Most dialysis patients have "intact" PTH (iPTH) levels between 65 and 450 pg/ml, which are poorly predictive of bone turnover state, making the iPTH test of limited value for bone turnover prediction. iPTH levels higher than 500 pg/ml can be observed in some dialysis patients with low bone turnover, while iPTH levels as low as 100 pg/ml have been found in patients with bone turnover above normal, indicating the standard second generation iPTH assay is not a reliable sole indicator of bone turnover. The whole PTH immunoradiometric assay, a third generation assay, uses a detection antibody that recognizes antigenic determinants at the extreme amino-terminal (1-4) end of the PTH molecule, making the assay specific for biologically active whole PTH-(1-84). Comparing results using the whole PTH and iPTH assays, the PTH-(7-84) level is indirectly determined and the PTH-(1-84)/PTH-(7-84) ratio can be calculated. It was shown that PTH-(7-84) inhibits the calcemic effect of PTH-(1-84) and its stimulatory effect on bone turnover. In the interpretation of results using the PTH-(1 84)/PTH-(7-84) ratio, it must be taken into consideration that second generation "intact" PTH assays have different cross-reactivity with PTH-(7-84). Therefore, when comparing or analyzing PTH-(1-84)/PTH-(7-84) ratios, the employed PTH assays must be identical. The whole PTH assay and the PTH-(1-84)/PTH-(7-84) ratio allow more meaningful interpretation of PTH trends, and offer a non-invasive means to more accurately diagnose bone disease in this population. PMID- 12779092 TI - A review of intravenous versus oral vitamin D hormone therapy in hemodialysis patients. AB - BACKGROUND/METHODS: We examined 21 clinical trials (16 articles and 5 abstracts) that compared intravenous (i.v.) and oral vitamin D analogs for the treatment of secondary hyperparathyroidism in hemodialysis patients. Nearly all the studies had severe limitations, especially study size. Only 2 studies utilized more than 25 patients per treatment arm; 11 of the 16 articles and 4 of the 5 abstracts had less than 15 patients per arm. Calcitriol and/or alpha-calcidol were studied in 20 trials of the 21 studies (15 of the 16 articles) while 1 article examined doxercalciferol. RESULTS: No difference of efficacy between i.v. and oral dosing was found in 10 of the 15 articles in which efficacy was assessed. The i.v. route provided significantly faster suppression of elevated parathyroid hormone (PTH) and/or a greater degree of suppression in 5 of 15 applicable articles, but in 2 of these 5 studies the i.v. dose was substantially greater than the oral dose. Side effects, chiefly hypercalcemia, were noted in half of the articles. Six of 9 articles with detailed results found no significant difference; only 2 found significantly increased hypercalcemia with oral dosing, and 1 found significantly increased hypercalcemia with i.v. dosing. Only 3 articles reported on hyperphosphatemia and no difference was found for mode of administration. One factor influencing 19 of the 21 comparisons was the use of oral doses that were therapeutically equivalent to about half or less the i.v. dose given the lower bioavailability of oral D hormones. One larger study (70 patients) that compared equipotent dosing of the 2 administration routes found 4 times more hypercalcemia using oral than i.v. dosing (p < 0.001). Another factor complicating interpretation is that the treatment periods were short, with half being 16 weeks or less and only 2 lasting 36 weeks. CONCLUSION: Conclusions about the comparative efficacy and safety of the 2 administration routes require larger studies of longer duration that utilize therapeutically equivalent doses. PMID- 12779093 TI - Serum potassium in the crush syndrome victims of the Marmara disaster. AB - BACKGROUND: Hyperkalemia is a major cause of mortality in the patients who suffer from crush syndrome in the aftermath of major earthquakes. The aim of this study is to investigate the frequency and effects of hyperkalemia in the 639 victims of catastrophic Marmara earthquake that struck northwestern Turkey, in August 1999. PATIENTS AND METHODS: Within the first week of disaster, questionnaires were sent to 35 reference hospitals that treated the victims. Information on serum potassium which was provided in 595 out of 639 questionnaires was submitted to analysis. RESULTS: In the patients who were admitted within the first 3 days of the disaster (n = 401) serum potassium was 5.4 +/- 1.3 mEq/l, which was higher than in those admitted thereafter (n = 171) (4.5 +/- 1.1 mEq/l) (p = 0.02). Considering the whole series, males (p = 0.01), patients needing dialysis support (p < 0.001) and non-survivors (p = 0.001) were characterized by higher serum potassium at admission. Seventy patients' serum potassium was above 7 mEq/l, while 22 patients were hypokalemic (< 3.5 mEq/l). Admission potassium correlated with many clinical and laboratory variables indicating the severity of the trauma, and a logistic regression model with clinical and laboratory parameters upon admission, revealed potassium as the most significant predictor of dialysis needs in the victims admitted within the first 3 days (p = 0.008, OR = 3.33). Among the victims who were admitted to hospitals 1 week after the disaster, 8 had serum potassium levels above 6.5 mEq/l; among 4 of them were complicated by hyperkalemia even higher than 7.5 mEq/l. These findings undeline the importance of hyperkalemia during clinical course. CONCLUSION: The most important and fatal medical complication in crush syndrome patients is hyperkalemia. Risk of fatal hyperkalemia continues even after hospitalization. Empirical therapy at the scene is indicated especially in male victims with severe soft tissue traumas. Early detection and treatment of hyperkalemia may improve the final outcome of renal disaster victims. PMID- 12779094 TI - Survival analysis of the factors affecting in mortality in injured patients requiring dialysis due to acute renal failure during the Marmara earthquake: survivors vs non-survivors. AB - BACKGROUND: We reviewed medical records of dialyzed patients admitted to our hospital after the Marmara earthquake and evaluated the factors affecting mortality in survivors and non-survivors according to the survival times. PATIENTS AND METHODS: Crush syndrome (CS) was diagnosed in 110 patients. Dialysis treatment was initiated in 60 patients; 21 of all died. The patients were divided into 2 groups which consisted of 39 survivors (Group A, 25 male, 14 female, mean age: 31 +/- 2.2 years) and 21 non-survivors (Group B, 9 male, 12 female, mean age: 27 +/- 3.0 years). Victims treated by any form of renal replacement therapy, including daily or intermittant hemodialysis and/or continuous venovenous hemodiafiltration. Clinical and laboratory findings were recorded regularly. Statistical analysis was performed with Kaplan-Meier method, log rank test and Cox regression analysis for the survival functions. RESULTS: APACHE II scores were 13.5 +/- 0.5 for Group A and 13 +/- 0.9 for Group B. Dialysis support was started to patients in Group A in a mean period of 2.8 +/- 0.2 days and in Group B in a mean period of 3.7 +/- 0.6 days after the earthquake (p > 0.05). The most frequent site of trauma was lower extremity (61.5%) and upper + lower extremities (23%) in Group A, and lower extremity (38.1%) and trunk + lower extremity (23.8%) in Group B. The frequencies of abdominal trauma, pelvic fracture and thoracic trauma in Group B were 23.8%, 19% and 14.2%, respectively. Multiple trauma was more frequent in Group B than in Group A (42.8% vs 2.5%). The rates of fasciotomy, amputation and surgery were similar in both groups. The frequency of sepsis was higher in non-survivors. In our center, the overall mortality rate was 8%, mortality rate in CS was 21% and in dialyzed patients it was 35%. Mortality was mainly associated with sepsis. Survival periods (52.3 +/- 4.0 days) in Group A were longer than in Group B (17.3 +/- 2.5 days). With Cox regression analysis, the parameters such as systolic hypotension on admission, female gender, high serum peak creatine kinase (> 20,000 U/l) and multiple trauma including thoracic and abdominal regions, were factors increasing risk of mortality. CONCLUSION: As a result, sepsis, multiple trauma and severe crush injury were the main factors increasing mortality risk in dialyzed injuries after the earthquake. PMID- 12779095 TI - Acute renal failure in the tropics: emerging trends from a tertiary care hospital in South India. AB - AIM: To study epidemiology, laboratory parameters, outcome and factors determining outcome of patients presenting with acute renal failure in our hospital. MATERIALS AND METHODS: A prospective study between December 1997 December 1999 in which all patients presenting with acute renal failure were included. Demographics, laboratory parameters, etiology, outcome and prognostic factors determining outcome were studied. RESULTS: 642 patients were part of the study. The predominant underlying cause for ARF was sepsis (153/642); overall mortality was 26.5%. Patients who had sepsis were admitted in the ICU, and patients with oliguria had poorer outcome. CONCLUSION: Sepsis continues to be the predominant cause of ARF. Overall mortality of our patients is better, because of the case mix, a large percentage of patients had acute gastroenteritis as a cause of CRF. PMID- 12779096 TI - Renal function in proteinuric glomerular diseases correlates to the changes in urine IgM excretion but not to the changes in the degree of albuminuria. AB - Renal function in proteinuric glomerular diseases correlates to the changes in urine IgM but not to the changes in the degree of albuminuria. BACKGROUND: Albuminuria is believed to correlate to the progression of renal failure in glomerular diseases. Nevertheless, many patients with glomerular disorders maintain their renal function despite persistent albuminuria. In previous studies, we found that the baseline urine excretion of IgM, rather than the degree of albuminuria, predicts the renal outcome in glomerulopathies. In the present study, we examine correlations between changes in the content and in the amount of urine proteins and renal survival during a follow-up time of 3.5 years. METHODS: An observational study of a mean of 44 (+/- 3.6) months was conducted in 37 proteinuric patients (21 males and 16 females) with biopsy-verified primary glomerular disease. The patients were subdivided, according to the findings at the end of the study, into 3 groups, 1 group with decreasing albuminuria (by more than 50%), 1 group with persisting albuminuria and low (< 0.04 mg/mmol creatinine) urinary IgM excretion and 1 group with persisting albuminuria and with high (> or = 0.04 mg/mmol) urinary IgM excretion. RESULTS: All patients that showed remission of albuminuria had also low IgM excretion at the end of the study. All these patients, except 1, maintained their renal function. Patients with persistent albuminuria and high urinary IgM excretion showed a decrease in the glomerular filtration rate (GFR) of a mean of 9.6 ml/min/year compared to a mean GFR increase by 1.5 ml/min/year in patients with low IgM excretion and the same degree of albuminuria (p < 0.01). Seven out of the 9 patients in the former group fall in GFR by more than 5 ml/min/year compared to only 1 of the 10 patients in the latter group. Furthermore, the GFR alterations that occurred during follow-up time correlated in a higher degree to the changes in urinary IgM excretion (r = 0.6, p < 0.01) than to the changes in the degree of albuminuria, (r = 0.4, p < 0.05). A stepwise regression analysis indicated that increased urine IgM excretion is a strong predictor of the GFR decline (r = 0.73, p < 0.001). CONCLUSION: High urinary IgM excretion correlates to decreased GFR in primary glomerular diseases regardless of the degree of albuminuria. In parallel, low urinary IgM excretion indicates beneficial prognosis in these diseases. Since IgM passes the glomerular barrier entirely through large shunts or defects in the glomerular capillary wall, decreased urine content of IgM might be considered as a sign of recovery in the glomerular damage. PMID- 12779097 TI - Genetic variations of the SLC7A9 gene: allele distribution of 13 polymorphic sites in German cystinuria patients and controls. AB - Cystinuria is a hereditary disorder of cystine and dibasic amino acid transport across the luminal membrane of renal tubules and intestine, resulting in recurrent nephrolithiasis. While mutations in the SLC3A1 gene cause type I cystinuria, patients with non-type I cystinuria carry mutations in the SLC7A9 gene. Both gene products form the renal amino acid transporter rBAT/b0,+AT affected in cystinuria. In the present study a total of 59 patients with different ethnic background were screened for sequence variations in SLC7A9, out of these 32 were of German origin. For determination of allele frequencies of detected polymorphisms, 58 healthy German controls were investigated. Molecular genetic analysis was performed using single-strand conformation polymorphism analysis, restriction assays and sequencing. Allele frequencies were analyzed statistically for the detected polymorphisms. In addition to the 6 already known variants we identified 7 new polymorphisms. Statistical analyses showed a significantly different distribution of alleles between German patients and German controls in case of the polymorphisms c. 147C>T (exon 2), c.386C>T (exon 3), IVS3+22T>G, c.584C>T (exon 4), c.610T>C (exon 4), c.692C>T (exon 5), c.852C>A (exon 6) and c.872C>T (exon 6). In summary, our results show that cystinuria is a complex disease which is not only caused by mutations in SLC7A9 and SLC3A1, but also influenced by other modifying factors such as variants in SLC7A9. PMID- 12779098 TI - Resting energy expenditure in chronic kidney disease: relationship with glomerular filtration rate. AB - BACKGROUND: The kidneys may account for up to 20% of resting energy expenditure (REE), but there is controversy regarding the relationship between severity of renal impairment and REE. Furthermore, reduction in protein intake is associated with declining glomerular filtration rate (GFR) that contributes to malnutrition. If a selective aversion to protein intake or a global reduction in food intake takes place is not known. METHODS: To define the relationship between REE and its relationship to GFR, we measured REE with indirect calorimetry, GFR with iothalamate clearance, and body composition with creatinine kinetics and skin fold measurements in 16 patients with mild to moderate renal failure due to type 2 diabetes mellitus (n = 12) or glomerulonephritides (n = 4). RESULTS: In univariate analysis, age, weight, height, body mass index, lean body mass, urine urea nitrogen but not hemoglobin or albumin correlated with REE. In multivariate analysis, lean body mass emerged as the strongest predictor of REE followed by GFR. A lower GFR was associated with a lower REE. Also, we found reduced REE, but not a fall in GFR, was associated with a fall in dietary protein intake. This supports the hypothesis that patients with CKD have no selective aversion to protein; reduction in dietary protein intake with progressive CKD is due to global reduction in food intake. CONCLUSIONS: We conclude that REE falls with declining renal function in patients with CKD. This is likely secondary to combination of reduced energy expenditure of kidneys, adaptation to decreased energy intake or alteration in cellular energy metabolism. PMID- 12779099 TI - Clinical significance of eosinophils in suspicious or borderline renal allograft biopsies. AB - AIMS: Renal allograft biopsies play a critical role in renal transplantation. Acute rejection characterized by tubulitis and intimitis is of primary concern. There is an association between eosinophilic infiltrates and irreversible acute rejection; however, the significance of eosinophils in biopsies that fall short of the diagnostic threshold for acute rejection has not been well studied. This report describes clinical course, treatment and long-term outcome of 5 transplant recipients with biopsy histology that showed borderline changes associated with eosinophilic infiltrates. METHODS: Clinical records were selected for review on the basis of biopsy histology satisfying the following criteria: presence of interstitial infiltrates with eosinophils, absence of definitive criteria for acute rejection and absence of findings suggestive of infection or cyclosporine toxicity. RESULTS: All identified biopsies occurred within the first month of transplantation, and histology showed varying degrees of patchy mononuclear cell infiltrates composed of lymphocytes, with eosinophilic infiltrates, but no evidence of acute rejection based on Banff criteria. These patients were taking trimethoprim-sulfamethoxazole and ranitidine at the time of biopsy. Serum creatinine returned to baseline levels in each case after stopping both drugs, and remained stable during the duration of follow-up without any documented episode of acute rejection. No patient received specific therapy for acute rejection. CONCLUSION: This report suggests that independent of decisions on treatment with high-dose steroids or anti-lymphocyte antibody preparations, the management algorithm should include stopping drugs associated with acute interstitial nephritis when non-diagnostic biopsies show eosinophilic infiltrates. PMID- 12779101 TI - The outcome of pregnancy among patients receiving hemodialysis treatment. AB - Pregnancy among women receiving hemodialysis as a renal replacement therapy is rare and the chance of a successful delivery is relatively low. In this article, we present 3 cases of women conceiving either before or after the initiation of hemodialysis. The various risks of each pregnancy and the outcome are discussed as well as the latest management guidelines for pregnant dialysis patients. PMID- 12779100 TI - Long-term evaluation of nutritional status using dual-energy X-ray absorptiometry in chronic hemodialysis patients. AB - BACKGROUND: Hemodialysis patients very often suffer from malnutrition with net loss of body muscle and fat stores. Since protein-calorie malnutrition has been considered to be highly associated with increased morbidity and mortality, it is important that the nutritional status is evaluated accurately and that long-term evaluation of the nutritional status is performed to obtain good outcome. PATIENTS AND METHODS: Forty-six patients (28 men and 18 women) undergoing maintenance hemodialysis were studied. Nutritional status of the patients was examined by dual-energy X-ray absorptiometry (DXA) and anthropometry measurements after a hemodialysis session and 4 years after the initial measurement. RESULTS: Percent fat, the ratio of body fat (BF) to weight, determined by DXA correlated positively with body mass index (BMI) calculated from weight and height. There was also a significant positive correlation between lean body mass (LBM) determined by DXA and arm muscle circumference (AMC) calculated using the anthropometric method. In the study period of 4 years, LBM and bone mineral content (BMC) observed at the end point were significantly lower than those at the start. In contrast, BF and %fat at the end point were significantly higher as compared with those at the initial measurement. CONCLUSION: These results indicate that DXA is a reliable method for body composition analysis in chronic hemodialysis patients. Because the detection of early alterations in body composition may provide an early indication of the development of malnutrition, serial evaluation of body composition using DXA should be valid for assessment of the nutritional status. PMID- 12779102 TI - Acute renal failure due to non-Hodgkin lymphoma infiltration of the kidneys detected by ultrasonography and confirmed by positron emission tomography. AB - Acute renal failure (ARF) as a consequence of non-Hodgkin lymphoma infiltration of the kidneys (LIK) is an uncommon complication of non-Hodgkin lymphoma. In literature, ARF due to LIK is reported in progressive disease. A case of non oliguric acute renal failure secondary to relapse of large B cell non-Hodgkin lymphoma primarily localized in the mediastinum is reported. LIK of both kidneys was diagnosed by ultrasonography, computer tomography scan and 18 fluorodeoxyglucose positron emission tomography. No other causes for renal failure were found. The prognosis of renal involvement in relapsed non-Hodgkin lymphoma is poor, as is demonstrated by our case. PMID- 12779104 TI - Weight loss, fever and a swollen elbow joint in a hemodialysis patient--a case of tuberculous arthritis. PMID- 12779103 TI - Neurotoxicity induced by Cefepime in a very old hemodialysis patient. AB - Neurotoxicity is an unusual complication of cephalosporin therapy. Only few cases of neurotoxicity induced by Cefepime have been described and probably the frequency of Cefepime-induced status epilepticus is underestimated. We report a case of an 82 year-old male, ESRD patient on chronic hemodialysis program affected by pneumonia, who received a treatment with intravenous Cefepime (1 g/day) and developed a seizure 4 days after the starting antibiotic therapy. Cefepime-induced neurotoxicity was suspected and its administration was immediately discontinued. In order to increase Cefepime clearance a hemodialysis session was urgently started and an improvement of his conscious level was observed. On the following day, after a second hemodialysis session his clinical condition and the status of neurotoxicity were completely recovered. The patient was discharged from the hospital in stable clinical condition one week later. At variance with the cases previously reported, the daily dose of Cefepime administrated to our patient was 50% lower and respected drug prescription dosage. Thus, we speculate on the hypothesis that advanced age of our patient and metabolic encephalopathy induced by chronic uremia made him more sensitive to the neurotoxicity induced by the drug. In conclusion, our case suggests that, in very old patients on long-term hemodialysis, it should be considered, to avoid neurotoxicity, to monitor the clinical neurological status, to use Cefepime at lower dosage than that allowed in patients with severe renal impairment (1 g/day) and, when possible, to evaluate Cefepime plasma levels. However, in these patients, other agents of the same class should be considered such as Cefotaxime and Ceftriaxone which are characterized by both an hepatic and renal excretion. In alternative to cephalosporins, antibiotics with the same action spectrum in the absence of neurological toxicity (i.e. Meropenem) should be recommended. PMID- 12779105 TI - Bartter's/Gitelman's syndrome: a model for the relationships between hypertension, angiotensin II, oxidative stress and remodeling. PMID- 12779106 TI - Dental anesthesiology--a most noble profession. PMID- 12779107 TI - Should a mucoadhesive patch (DentiPatch) be used for gingival anesthesia in children? AB - A local anesthetic-impregnated mucosal adhesive patch (DentiPatch) was compared with topical anesthetic (Hurricaine Dry Handle Swab) for gingival anesthesia before rubber dam clamp placement in children. Twenty-eight children needing sealants on their posterior teeth were enrolled in this study. Topical anesthesia was provided using either the mucoadhesive patch (20% lidocaine) or topical anesthetic (20% benzocaine). Subjects were randomized using a split mouth model. Either the patch or topical anesthetic was applied to the gingiva for 5 minutes or 1 minute, respectively. Subjects used a visual analog scale to describe their pain during the procedure. Linear regression and mixed linear models were used for data analysis. The visual analog scale results (pain scores) showed no significant difference between treatments. The mean per-child patch-sticking fraction was 29.7%. Patch adherence to oral mucosa increased with age in girls (P = .0045), but not in boys. The DentiPatch is as effective as, although not superior to, the Hurricaine Dry Handle Swab for gingival anesthesia before rubber dam clamp placement in children. These study results would not support the use of the DentiPatch for gingival anesthesia in children because of poor adherence to oral mucosa and the extra time necessary to apply and retain the device. PMID- 12779108 TI - The relationship of placement accuracy and insertion times for the laryngeal mask airway to the training of inexperienced dental students. AB - Any health care professional can be faced with a medical emergency in which the patient needs ventilatory support. Bag-valve-mask ventilation with the assistance of an oropharyngeal airway that uses 100% oxygen is currently the preferred method for artificial ventilation. This procedure is generally performed ineffectively by most dentists inexperienced in airway management. We examined whether a short and simple period of training by dental students inexperienced in airway management would increase the speed and accuracy of the placement of the laryngeal mask airway (LMA), which may be a superior airway device to the bag valve-mask and oropharyngeal airway. Thirty-five dental students inexperienced in airway management were divided into 3 groups. The first group received only a demonstration on how to use the LMA. The second and third groups received the demonstration plus practiced inserting the LMA 5 and 10 times, respectively. A dental anesthesiologist graded the placement of the LMA with a tracheobroncho fiberscope (fiberoptic bronchoscope). Those who practiced inserting the LMA 5 times faired better than those who received no training; however, those who practiced 10 times did not do any better than the second group. The LMA can be inserted rapidly and effectively by dentists inexperienced in airway management after a short period of simple training that may be critical when personnel experienced in intubation are not readily available. PMID- 12779109 TI - A comparison study between ketamine and ketamine-promethazine combination for oral sedation in pediatric dental patients. AB - This study compared the incidence of vomiting and the sedative effectiveness of ketamine to a ketamine-prornethazine combination in pediatric dental patients. Twenty-two patients with American Society of Anesthesiologists' classification I physical status who were between the ages of 21 and 43 months were randomly divided into 2 groups. The control group received 10 mg/kg of ketamine orally, whereas the experimental group received 10 mg/kg of ketamine and 1.1 mg/kg of promethazine orally. Nitrous oxide in oxygen was supplemented between 35 and 50%. Each patient received 1 or 2 quadrants of restoration by one operator. Heart rate, blood pressure, and oxygen saturation were monitored and recorded during the treatment. Crying, alertness, movement, and overall general behavior were rated using the scale by Houpt et al. A dentist-anesthesiologist conducted the vital sign monitoring and behavioral assessment. Ketamine combined with promethazine eliminated the incidence of vomiting. A 2 x 2 chi-square contingency table showed a statistical difference between the 2 groups at P < .05 (control group, 27%; experimental group, 0%). Ketamine alone yielded better sedations than the combined agents as shown by the Mann-Whitney U statistical analysis (P < .05). Ketamine and a ketamine-promethazine combination are effective in the sedation of pediatric dental patients. PMID- 12779110 TI - Good isn't good enough anymore: upgrading training and provider evaluations. PMID- 12779111 TI - Pharmacokinetics of oral tramadol drops for postoperative pain relief in children aged 4 to 7 years--a pilot study. AB - Tramadol hydrochloride is an analgesic with mu receptor activity suitable for administration to children as oral drops. As the serum concentration profile and pharmacokinetic parameters in young children are not known via this route, we studied 24 healthy ASA 1 children to determine those parameters. The children's mean age was 5.3 +/- 1.1 years and their mean weight was 17.8 +/- 3.1 kg. They underwent general anesthesia with sevoflurane for dental surgery. The mean duration of anesthesia was 27.9 +/- 10.1 minutes. Tramadol 1.5 mg/kg (this dose was chosen because we have previously shown it to be effective in providing analgesia following pediatric dental surgery) was administered as oral drops 30 minutes before anesthesia. Venous blood samples were taken following the tramadol at 30-minute intervals for 4 hours, every 2 hours for 6 hours, and every 4 hours for 12 hours. The samples were centrifuged and the serum stored at -20 degrees C, and nonstereoselective gas chromatography was used to determine the concentration of (+) and (-) tramadol enantiomers plus their o-demethyltramadol (M1) metabolite concentrations. The tramadol absorption was rapid, the maximum measured serum concentration present occurring before the first sample at 30 minutes. That first sample had a concentration of 352 +/- 83.4 ng/mL. The concentration remained above the 100 ng/mL analgesic level until 6.8 +/- 0.9 hours. The elimination half life was 3.6 +/- 1.1 hours, the serum clearance 5.6 +/- 2.7 mL/kg/min, and the volume of distribution 4.1 +/- 1.2 L/kg. The (+) enantiomer concentration was 14.2 +/- 4.9% greater than that of the (-) enantiomer. The M1 metabolites had a ( ) enantiomer concentration 92.3 +/- 75.1% greater than the (+) enantiomer. From the peak concentration at 4.5 +/- 1.5 hours, the concentration of the metabolite was approximately one third that of the parent drug. The M1 elimination half-life was 5.8 +/- 1.7 hours. Apart from the rapid rise in the serum concentration, these kinetic parameters are similar to those seen in healthy young adults. The concentration profile supports an effective clinical duration in the region of 7 hours. PMID- 12779112 TI - Investigation of previously reported mucosal swellings after injection with Citanest Forte. AB - The purpose of this study was to determine the reason for an apparent increase in the number of mucosal swellings after maxillary infiltration with Citanest Forte (prilocaine HCl 4% solution with epinephrine 1:200,000), 2 years after its introduction in 1971 by Astra Pharmaceutical Co (now AstraZeneca) in the United States. Approximately 70% of these reported reactions were from California, where less than 11% of all cartridges were sold. Comparison with New York State, with 27% of total sales but less than 1% of the reactions, suggested that possible differences in practice characteristics were responsible for the swellings. On the basis of the Bureau of Economic Research and Statistics Survey of Dental Practice, dentists in the Far West (eg, California) were found to schedule appointments with a median length of approximately twice that of their Mid-East colleagues, the implication being that more anesthetic solution was injected per office visit. Follow-up telephone interviews of dentists reporting such reactions at that time verified that they administered more than the recommended 1.8-mL dose. The most important epidemiologic information was that prilocaine HCl 4% solution with epinephrine 1:200,000 had been on sale in Canada 4 years before it was introduced in the US market, with little or no evidence of drug-related effects. Comparison of the US and Canadian prilocaine HCl with epinephrine 1:200,000 specifications revealed that NaCl was added to an already hypertonic prilocaine solution in the US but not in Canada. Comparison of the responses to intradermal injection of US and Canadian prilocaine solutions into the backs of rabbits with follow-up studies of dose-related NaCl injections demonstrated that the added NaCl was responsible for the onset and duration of irritation from the initially marketed US Citanest solutions. PMID- 12779113 TI - Clinical recovery time from conscious sedation for dental outpatients. AB - For dental outpatients undergoing conscious sedation, recovery from sedation must be sufficient to allow safe discharge home, and many researchers have defined "recovery time" as the time until the patient was permitted to return home after the end of dental treatment. But it is frequently observed that patients remain in the clinic after receiving permission to go home. The present study investigated "clinical recovery time," which is defined as the time until discharge from the clinic after a dental procedure. We analyzed data from 61 outpatients who had received dental treatment under conscious sedation at the Hiroshima University Dental Hospital between January 1998 and December 2000 (nitrous oxide-oxygen inhalation sedation [n = 35], intravenous sedation with midazolam [n = 10], intravenous sedation with propofol [n = 16]). We found that the median clinical recovery time was 40 minutes after nitrous oxide-oxygen sedation, 80 minutes after midazolam sedation, and 52 minutes after propofol sedation. The clinical recovery time was about twice as long as the recovery time described in previous studies. In a comparison of the sedation methods, clinical recovery time differed (P = .0008), being longer in the midazolam sedation group than in the nitrous oxide-oxygen sedation group (P = .018). These results suggest the need for changes in treatment planning for dental outpatients undergoing conscious sedation. PMID- 12779114 TI - Paradoxical reactions to benzodiazepines in intravenous sedation: a report of 2 cases and review of the literature. AB - Paradoxical reactions to benzodiazepines have been thoroughly reported since the introduction of this type of drug. The mechanism of benzodiazepine action is through the gamma-aminobutyric acid receptors. Properties of benzodiazepine include sedation, anxiolysis, amnesia, anticonvulsion, and muscle relaxation. Unfortunately, adverse paradoxical reactions can be stimulated by benzodiazepines and are difficult to predict and diagnose. Two cases of paradoxical reactions associated with the use of intravenous midazolam are presented, and the management of this complication and its different etiologies are reviewed. The relationship of the paradoxical reaction to alteration of the cholinergic homeostasis, serotonin levels, the role of genetics, and gamma-aminobutyric acid receptor configuration is discussed. PMID- 12779115 TI - Sexy transgenes: the impact of gene transfer and gene inactivation technologies on the understanding of mammalian sex determination. AB - Amongst the various developmental pathways ending in a sound mammal, sex determination presents the peculiarity of a choice between two equally viable options: female or male. Therefore, destroying a 'male-determining gene' or a 'female-determining gene' should generally not be lethal. Genetic sex determination is divided into two consecutive steps: construction of the bipotential gonad, and then sex determination per se. The genes involved in the first step are in fact involved in the development of various body compartments, and their mutation is generally far from innocuous. From transgenic and inactivation studies carried out on the laboratory mouse, a complete picture of the two steps is beginning to emerge, where the gonad itself and the necessary ducts are shown to evolve in a very coordinate way, with well-defined sex specificities. Compared with testis determination, the ovarian side of the picture is still relatively empty, but this situation can change rapidly as candidate ovarian genes for inactivation studies are beginning to be identified. PMID- 12779116 TI - The promoter of a metallothionein-like gene from the tropical tree Casuarina glauca is active in both annual dicotyledonous and monocotyledonous plants. AB - A chimeric gene consisting of the beta-glucuronidase (gusA) reporter gene under the control of the metallothionein-like promoter cgMT1 from the tropical tree Casuarina glauca was introduced into Nicotiana tabacum via Agrobacterium tumefaciens and into Oryza sativa by particle bombardment. The strongest histochemical staining for GUS activity was observed in the root system of the transgenic plants, and especially in lateral roots. In contrast, a relatively low level of reporter gene expression was seen in the aerial tissues and GUS staining was located mainly in the plant vascular system. The average ratio of GUS activity between root and leaf was found to be 13:1 in tobacco and 1.5:1 in rice. The pattern of cgMT1 promoter activity in floral organs was found to be different in tobacco and rice. High levels of gusA gene expression were detected in the ovules, pollen grains and tapetum, whereas in rice PcgMT1 directs expression to the vascular system of the floral organs. These results suggest that PcgMT1 is potentially useful in molecular breeding to express genes of interest whose products are preferentially needed in roots. PMID- 12779117 TI - Transgenic mice expressing recombinant human protein C exhibit defects in lactation and impaired mammary gland development. AB - To determine if the production of recombinant human protein C (rHPC) could be increased in milk, we created two lines of mice homozygous for the mouse whey acidic protein (WAP)/human protein C (HPC) transgene. Females of both lines had normal growth, activity and fertility, but failed to lactate normally and were unable to raise litters. Histological analyses of mammary glands from lactating homozygous females showed barely distended alveoli filled with dense-staining milk. Epithelial cells within these alveoli had distinct, centrally located nuclei and contained intracellular lipid droplets. Hemizygous animals derived from these lines were able to lactate and raised normal sized litters. Northern blot analysis showed that the 6.4 homozygous (6.4H) line expressed the transgene at higher levels then corresponding hemizygous (6.4) animals, but the 4.2 homozygous (4.2H) line expressed the transgene at lower levels than the 4.2 hemizygous line. The 6.4H line also had increased rHPC levels in the milk as revealed by western blot analysis. The 4.2H, 6.4, and 6.4H lines showed decreased and/or delayed expression of WAP, beta-casein, and alpha-lactalbumin mRNA's compared to wild type animals during lactogenesis. The 4.2 line showed decreased mRNA expression for beta-casein and alpha-lactalbumin, but normal or higher expression of WAP during lactogenesis. Elevated levels of some proteins were detected in the milk of transgenic mice. From these results, it is concluded that expression of rHPC induced a lactational phenotype that involves abnormal morphological, biochemical, and functional differentiation of mammary epithelial cells. However, the induction of this phenotype does not appear to be directly related to the level of rHPC mRNA expression, thus suggesting that the basis of this phenotype may involve secondary, rather than primary, effects of rHPC on mammary gland development. PMID- 12779118 TI - Monitoring the spread of recombinant DNA from field plots with transgenic sugar beet plants by PCR and natural transformation of Pseudomonas stutzeri. AB - Previous studies had shown that recombinant DNA can be detected for several months in soil after the deposition of litter from transgenic (tg) plants. Here we show by PCR monitoring of field releases of tg sugar beet plants that during the growth of the plants the soil close to the plants and also plant material contains recombinant DNA, in the form of extracellular molecules. Surprisingly, the monitoring also revealed the presence of tg DNA in many field plots (30-70%) in which tg plants were never grown. These studies and the further monitoring during other tg sugar beet release experiments by PCR and a novel bioassay (measuring the transforming potential of recombinant DNA for Pseudomonas stutzeri) indicated that recombinant DNA was only detectable in the surface soil of field plots and their vicinity where flowering of the tg beet plants was allowed. Recombinant DNA was found in soil at a distance of 50 m from pollen producing plants surrounded by a strip with hemp plants as a containment regime. It is concluded that recombinant DNA is deposited in soil during the growth of tg sugar beets and that a major mechanism of recombinant DNA spread in the environment is the dispersal of pollen which allows recombinant DNA to persist in the field plot for at least a year. PMID- 12779119 TI - Analysis of trans-silencing interactions using transcriptional silencers of varying strength and targets with and without flanking nuclear matrix attachment regions. AB - We investigated the effect of the Rb7 matrix attachment region (MAR) on trans silencing in tobacco plants, comparing the effects of three transgene silencer loci on ten target loci. Two of the silencer loci, C40 and C190, contain complex and rearranged transgene arrays consisting of 35S:GUS or NOS:NPTII containing plasmids. The third silencer locus, V271, was previously characterized as a complex locus containing rearranged 35S:RiN sequences. Each of these silencers can reduce 35S promoter-driven expression at other loci, albeit with varying efficiencies. The presence of MARs at a target locus does not prevent trans silencing by the V271 silencer. However, four of seven MAR-containing loci were at least partially resistant to silencing by the C40 and C190 loci. One MAR locus was unaffected by C40, our weakest silencer, and three were silenced only when the silencer locus was maternally inherited. Silencing is progressive in the F1 and F2 generations; two days after germination there is little or no difference between seedlings derived from crosses to silencing or control lines, but seedlings containing silencer loci slowly lose expression during subsequent development. These observations are compatible with the hypothesis that a product of the silencer locus must accumulate before unlinked loci can be affected. However, our silencer loci are themselves silenced for GUS transcription, and coding region homology is not required for their effects on target loci. Our results are consistent with a model in which transcriptional silencing is triggered by transcription of sequences during the early stages of embryo or seedling development. PMID- 12779120 TI - Tissue specific high level expression of a full length human endogenous retrovirus genome transgene, HERV-R, under control of its own promoter in rats. AB - Human endogenous retrovirus-R (HERV-R) is one of a full length HERV with a long open reading frame in the env region. The env transcripts are expressed in various human tissues. To investigate the biological role of HERV-R in vivo, we established two lines of transgenic rats carrying a full sequence of HERV-R under control of its own long terminal repeat (LTR) promoter. One line with tandem integration of multiple copies of the transgene expressed HERV-R mRNA in various organs with different expression levels and relatively higher in Harderian and submandibular salivary glands. In another line, the transgene was integrated as a single copy in a haploid and the expression was detected only in Harderian and submandibular salivary glands. In the placenta, one of the tissues with high levels of the HERV-R expression in humans, the transcription was evident starting the 12th day after gestation. A rabbit antiserum against synthetic peptides corresponding with the HERV-R env gene sequence led to detection of an 85 kDa product as a glycoprotein in the Harderian glands. While no pathological significance was observed in either line, the transgenic rat may prove to be a suitable model for analyzing the role of HERV-R function in vivo. PMID- 12779121 TI - The sweet pepper ferredoxin-like protein (pflp) conferred resistance against soft rot disease in Oncidium orchid. AB - Genetic engineering to date has not been used to introduce disease resistance genes into the orchid gene pool. The ferredoxin-like protein gene originally isolated from sweet pepper is thought to function as a natural defense against infection due to its antimicrobial properties. Hence it was reasoned that introduction of this gene might produce Oncidium plants resistant to Erwinia carotovora, the causal agent for the soft rot disease. An expression vector containing sweet pepper ferredoxin-like protein (pflp) cDNA, hph and gusA coding sequence was successfully transformed into protocorm-like bodies (PLBs) of Oncidium orchid, using Agrobacterium tumefaciens strain EHA105. A total of 17 independent transgenic orchid lines was obtained, out of which six transgenic lines (beta-glucuronidase (GUS) positive) were randomly selected and confirmed by Southern, northern and western blot analyses. A bioassay was conducted on the transgenic lines. Transgenic plants showed enhanced resistance to E. carotovora, even when the entire plant was challenged with the pathogen. Our results suggest that pflp may be an extremely useful gene for genetic engineering strategies in orchids to confer resistance against soft rot disease. PMID- 12779123 TI - Tritrophic choice experiments with bt plants, the diamondback moth (Plutella xylostella) and the parasitoid Cotesia plutellae. AB - Parasitoids are important natural enemies of many pest species and are used extensively in biological and integrated control programmes. Crop plants transformed to express toxin genes derived from Bacillus thuringiensis (Bt) provide high levels of resistance to certain pest species, which is likely to have consequent effects on parasitoids specialising on such pests. A better understanding of the interaction between transgenic plants, pests and parasitoids is important to limit disruption of biological control and to provide background knowledge essential for implementing measures for the conservation of parasitoid populations. It is also essential for investigations into the potential role of parasitoids in delaying the build-up of Bt-resistant pest populations. The diamondback moth (Plutella xylostella), a major pest of brassica crops, is normally highly susceptible to a range of Bt toxins. However, extensive use of microbial Bt sprays has led to the selection of resistance to Bt toxins in P. xylostella. Cotesia plutellae is an important endoparasitoid of P. xylostella larvae. Although unable to survive in Bt-susceptible P. xylostella larvae on highly resistant Bt oilseed rape plants due to premature host mortality, C. plutellae is able to complete its larval development in Bt-resistant P. xylostella larvae. Experiments of parasitoid flight and foraging behaviour presented in this paper showed that adult C. plutellae females do not distinguish between Bt and wildtype oilseed rape plants, and are more attracted to Bt plants damaged by Bt-resistant hosts than by susceptible hosts. This stronger attraction to Bt plants damaged by resistant hosts was due to more extensive feeding damage. Population scale experiments with mixtures of Bt and wildtype plants demonstrated that the parasitoid is as effective in controlling Bt-resistant P. xylostella larvae on Bt plants as on wildtype plants. In these experiments equal or higher numbers of parasitoid adults emerged per transgenic as per wildtype plant. The implications for integrated pest management and the evolution of resistance to Bt in P. xylostella are discussed. PMID- 12779122 TI - Transgenic and tissue culture analyses of the muscle creatine kinase enhancer Trex control element in skeletal and cardiac muscle indicate differences in gene expression between muscle types. AB - The muscle creatine kinase (MCK) gene is expressed at high levels only in differentiated skeletal and cardiac muscle. The activity of the cloned enhancer promoter has previously been shown to be dependent on the Trex element which is specifically bound by a yet unidentified nuclear factor, TrexBF. We have further characterized the function of the Trex site by comparing wild-type and Trex mutated MCK transgenes in five mouse skeletal muscles: quadriceps, extensor digitorum longus (EDL), soleus, diaphragm, and distal tongue, as well as in heart ventricular muscle. Several types of statistical analysis including analysis of variance (ANOVA) and rank sum tests were used to compare expression between muscle types and between constructs. Upon mutation of the Trex site, median transgene expression levels decreased 3- to 120-fold in the muscles examined, with statistically significant differences in all muscles except the EDL. Expression in the largely slow soleus muscle was more affected than in the EDL, and expression in the distal tongue and diaphragm muscles was affected more than in soleus. Median expression of the transgene in ventricle decreased about 18 fold upon Trex mutation. Transfections into neonatal rat myocardiocytes confirmed the importance of the Trex site for MCK enhancer activity in heart muscle, but the effect is larger in transgenic mice than in cultured cells. PMID- 12779124 TI - Expression of the bovine growth hormone alters the root morphology in transgenic tobacco plants. AB - The bovine growth hormone (bGH) is a natural peptide hormone that controls the differentiation, growth and metabolism, and is produced in the pituitary gland of cows. For the production of bGH from plants, two different bgh clones, of which the pGAbGH1 contaions only mature peptide sequences and the pGAbGH15 contains signal sequences and the first intron, as well as mature peptide sequences, were used. Those bghs under the control of the CaMV 35S promoter and NOS terminator were introduced to tobacco plants via Agrobacterium tumefaciens-mediated transformation. By PCR analyses using bgh and nptII specific primers, 17 and 21 putative transformants were respectively selected from pGAbGH1- and pGAbGH15 transformed tobacco plants. Northern blot analysis showed that the most of the transgenic lines expressed the bgh mRNA. Western blot analysis revealed that the pGAbGH1-transformed tobaccos produced recombinant bGH, but pGAbGH15-transformed ones did not produce the protein. Interestingly, some morphological changes were observed in the roots of transgenic tobacco plants. The transgenic tobacco plants had thick and short roots containing few root hairs in contrast to the non transformed wild type plants. PMID- 12779125 TI - Placental anomalies and fetal loss in mice, after administration of doxycycline in food for tet-system activation. AB - During the course of a study aiming to obtain a tetracycline (Tet)-inducible transgene expression restricted to the placenta, we have observed a toxicity of doxycycline (dox) given in the food at doses of 2.5-10 mg/g to pregnant mice from two different inbred strains. During the second half of gestation, dox-fed non transgenic mice presented placental anomalies and impaired fetal development proportional to the dose of antibiotic. Thus, dox administered in commonly used food doses can have an adverse effect on pregnancy. These observations are important for studies of placental or fetal development using inducible gene promoters. PMID- 12779126 TI - The production of transgenic Scots pine (Pinus sylvestris L.) via the application of transformed pollen in controlled crossings. AB - The study demonstrates the production of a transgenic Scots pine (Pinus sylvestris L.) seedling through the application of transformed pollen in controlled crossings. The pollen lots were transformed by particle bombardment, resulting in transient transformation frequencies varying from 15 to 49% of the germinated pollen grains, and bombarded pollen was used to pollinate megasporangiate strobili. Progeny was screened by histochemical, GUS assays, and selected seedlings were further analysed by PCR. PCR amplification revealed the presence of both the nptII and gusA genes in one seedling (23/237). Results were confirmed by Southern blot analysis. The morphology and growth of this transgenic seedling was normal. Although the transformation frequency of recovered plants was very low (1/14999), the present protocol suggests that production of transgenic Scots pine is possible without the use of any tissue culture methods or the involvement of marker genes, for selection of transformants. PMID- 12779127 TI - From emerging viruses to cardiovascular disease: preventive measures matter. PMID- 12779128 TI - Whither arthroscopic treatment for osteoarthritis of the knee? PMID- 12779129 TI - When to wean from a ventilator: an evidence-based strategy. AB - It is often unclear when and how to wean patients from mechanical ventilation. We have devised an evidence-based protocol in which patients undergo a 30-minute trial of spontaneous breathing with a T tube or pressure support of 7 cm H2O. Those who can tolerate the trial are extubated, while those who cannot are reconnected to mechanical ventilation but undergo another trial every day until they can be extubated. More study is needed to improve the criteria to predict successful spontaneous breathing and extubation, and to clarify the role of non invasive ventilation to avoid reintubation. PMID- 12779131 TI - Severe acute respiratory syndrome (SARS): update on a moving target. PMID- 12779130 TI - A perspective on the study of Moseley et al: questioning the value of arthroscopic knee surgery for osteoarthritis. AB - Arthroscopy for degenerative conditions of the knee is among the most commonly employed orthopedic procedures, but its effectiveness (like the effectiveness of many surgical operations) has never been proven in prospective trials. Moreover, the precise mechanism by which arthroscopy improves the course of degenerative conditions of the knee has not been established conclusively. Moseley et al performed a double-blinded, randomized, placebo-controlled trial to compare the effectiveness of arthroscopic lavage and arthroscopic debridement vs a sham procedure. Data regarding pain and function were obtained at multiple time points over a 2-year period. The authors found that all three treatment groups fared equally: each reported subjective symptomatic relief, but no objective improvement in function was noted in any of the groups. These data suggest that the benefits of arthroscopy for the treatment of osteoarthritis of the knee is to provide subjective pain relief, and that the means by which arthroscopy provides this benefit is via a placebo effect. PMID- 12779132 TI - A 51-year-old woman with debilitating cramps 12 years after bariatric surgery. PMID- 12779133 TI - Fecal incontinence in elderly patients: common, treatable, yet often undiagnosed. AB - It is important for primary care physicians to take fecal incontinence seriously and not dismiss it as a normal part of aging. Elderly patients may be reluctant to admit fecal incontinence, so clinicians need to ask about it. Two of the most common causes are fecal impaction (especially in nursing home patients) and rectosphincter dysfunction in people with diabetes. PMID- 12779134 TI - West Nile fever: lessons from the 2002 season. AB - West Nile fever has now spread to much of the United States. This disease can be diagnosed using one of several laboratory tests, notably an immunoglobulin M enzyme-linked immunosorbent assay. It can cause devastating neurologic damage, including an unusual polio-like syndrome. Magnetic resonance imaging is an important imaging tool in such patients. Treatment is largely supportive, although antiviral agents are under investigation. PMID- 12779135 TI - Patient information. Reducing your risk of West Nile virus. PMID- 12779136 TI - New guidelines for occupational exposure to blood-borne viruses. AB - The US Public Health Service recently updated its guidelines for managing health care workers exposed to blood or other body fluids that might contain blood-borne viruses. The update addresses, among other things, timely administration of hepatitis B immune globulin and hepatitis B vaccine, appropriate testing for hepatitis C exposure, and new information on prophylaxis after exposure to human immunodeficiency virus (HIV). PMID- 12779137 TI - Clopidogrel and thrombotic thrombocytopenic purpura: no clear case for causality. AB - Although the antiplatelet agent clopidogrel is associated with an increased incidence of thrombotic thrombocytopenic purpura (TTP), available evidence is not sufficient to establish or disprove a causal relationship. We review and assess the evidence and case reports linking clopidogrel use with TTP and discuss how to approach the risk of TTP in patients taking clopidogrel. PMID- 12779138 TI - Calcific aortic stenosis: another face of atherosclerosis? AB - Calcific aortic stenosis is not the result of decades of wear and tear alone. Rather, it is being increasingly recognized as an inflammatory, atheromatous, and potentially modifiable disease. Exciting new research suggests that medical therapies may soon be available to retard its progression and reduce the need for surgery. PMID- 12779139 TI - Equivalent volume-averaged light scattering behavior of randomly inhomogeneous dielectric spheres in the resonant range. AB - Finite-difference time-domain numerical experiments and supporting analyses demonstrate that the spectral dependence of the total scattering cross sections of randomly inhomogeneous dielectric spheres of sizes in the resonant range closely resemble those of their homogeneous counterparts that have a volume averaged refractive index. This result holds even for the extreme case in which the refractive index within an inhomogeneous sphere varies randomly over the range 1.0-2.0. PMID- 12779140 TI - Two-photon excited intramolecular energy transfer and light-harvesting effect in novel dendritic systems. AB - Two-photon absorption excited intramolecular energy transfer and light-harvesting effects are demonstrated in three novel dendritic systems. These systems contain both an antenna structure that can effectively absorb two-photon energy at approximately 800 nm and emit fluorescence at approximately 515 nm and a core moiety that can absorb one-photon energy at approximately 520 nm and emit at approximately 590 nm. Covalently combining the core and antenna functionalities intrinsically changes the optical behavior of the component pieces. The two photon energy absorbed by the antenna structure is resonantly transferred to the core, where the core's emission intensity is enhanced by 8, 20, and 34 times for the three dendritic systems. PMID- 12779141 TI - Phase pupil functions for reduction of defocus and spherical aberrations. AB - Radially symmetric pupil plane phase retardation functions are derived that extend focal depth and alleviate third-order spherical aberration (SA) effects. The radial symmetry of these functions means that they can be more conveniently manufactured by use of traditional techniques such as diamond machining than previously reported filters with rectangular symmetry. The method employs minimization of the variation of Strehl ratio with defocus, W20, and SA, W40. The performance of the derived phase filters is illustrated by comparison with standard optical systems and with previously reported phase filters. PMID- 12779142 TI - Summing Lax series for nonparaxial beam propagation. AB - It is shown that the approach proposed by Lax et al. [Phys. Rev. A 11, 1365 (1975)] for studying the propagation of an electromagnetic beam beyond the paraxial approximation can be efficiently employed evenwhen the beam under consideration presents a very nonparaxial character. The method that we present consists of applying a nonlinear resummation scheme, the so-called delta transformation, to the divergent perturbative series arising from the Lax scheme. Numerical results pertinent to the evaluation of transverse and longitudinal components of the electric field are presented for the particular case of vectorial Gaussian beams, showing the effectiveness of the method. PMID- 12779143 TI - Group-delay ripple correction in chirped fiber Bragg gratings. AB - Group-delay ripple (GDR) introduced by systematic and random errors in chirped fiber Bragg grating fabrication is the most significant impediment to application of these devices in optical communication systems. We suggest and demonstrate a novel iterative procedure for GDR correction by subsequent UV exposure by use of a simple solution of the inverse problem for the coupled-wave equation. Our method is partly based but does not fully rely on the accuracy of this solution. In the experiment we achieved substantial reduction of the low-frequency group delay ripple, from +/- 15 to +/- 2 ps, which resulted in dramatic improvement of the optical signal-to-noise-ratio system penalty, from 7 to less than 1 dB, for a chirped fiber Bragg grating used as a dispersion compensator in a 40-Gbit/s carrier-suppressed return-to-zero system. PMID- 12779144 TI - Suppression of beating noise of narrow-linewidth erbium-doped fiber ring lasers by use of a semiconductor optical amplifier. AB - Beating noise in narrow-linewidth erbium-doped fiber ring lasers puts severe limitations on applications of the lasers. We demonstrate the suppression of beating noise in fiber ring lasers by using a semiconductor optical amplifier in the laser cavity, which acts as a high-pass filter. Two different ring structures are presented as examples to demonstrate this beating noise suppression. PMID- 12779145 TI - Doped photonic bandgap fibers for short-wavelength nonlinear devices. AB - Microstructured photonic bandgap fibers with a doped honeycomb cladding structure and the guiding defect defined by the absence of doping are proposed as nonlinear optical fibers for short wavelengths. It is shown that zero-dispersion wavelengths below 500 nm and corresponding effective areas of 1-2 microm2 can be obtained if structures with interhole distances near 600 nm can be fabricated. The cutoff wavelength for guidance of second-order modes can be controlled by variation of the radius and index contrast of the doped regions. PMID- 12779146 TI - Strong phase-controlled fiber Bragg gratings for dispersion compensation. AB - Dispersion-compensating fiber Bragg gratings with approximately 99.9% reflectivity that are made by continuous apodization and phase control are demonstrated. These strong dispersion-compensating gratings provide precision second-order, third-order, or even more complex dispersion compensation, as well as sufficient transmission isolation to be used at add-drop stages without additional filtering. A 99.84% grating with a constant approximately 700-ps/nm dispersion and a 99.94% grating with dispersion varying linearly from 1000 to 1000 ps/nm are demonstrated. PMID- 12779147 TI - Nondestructive imaging of a type I optical fiber Bragg grating. AB - Nondestructive images of refractive-index variation within a type I fiber Bragg grating have been recorded by the differential interference contrast imaging technique. The images reveal detailed structure within the fiber core that is consistent with the formation of Talbot planes in the diffraction pattern behind the phase mask that had been used to fabricate the grating. PMID- 12779148 TI - Total-internal-reflection mode in holographic polymer dispersed liquid crystals. AB - We have developed a holographic polymer dispersed liquid-crystal device based on total internal reflection (TIR). In the field-off state, the holographic planes are slanted such that the angle of the diffraction beam is beyond the TIR angle of the glass substrate. The beam is trapped in the sample and propagates to the edge. In the field-on state, the holographic grating is erased and the incident beam propagates directly through the sample. Contrast ratios of 10 dB, submillisecond response times, and an index modulation of 0.042 at 1550 nm are reported. PMID- 12779149 TI - Optimum holographic optical element lens recorded by visible laser beams for an infrared two-dimensional vertical-cavity surface-emitting laser array. AB - We have designed an optimum holographic optical element (HOE) lens recorded by visible laser beams for an infrared two-dimensional vertical-cavity surface emitting laser (VCSEL) array. The hologram computer-aided design tool that we have developed is used for the optimum design. The optimum HOE has both high light efficiency and a small amount of aberration. An Nd:YAG laser operated at 532 nm and an 8 x 8 VCSEL array operated at 850 nm are used for HOE recording and reconstruction, respectively. The designed lens is experimentally demonstrated, and the experimental results of the lens almost agree with the numerical results. PMID- 12779150 TI - Experimental validation of Fourier-transform wave-front reconstruction at the Palomar Observatory. AB - Wave-front reconstruction with use of the Fourier transform has been validated through theory and simulation. This method provides a dramatic reduction in computational costs for large adaptive (AO) systems. Because such a reconstructor can be expressed as a matrix, it can be used as an alternative in a matrix-based AO control system. This was done with the Palomar Observatory AO system on the 200-in. Hale telescope. Results of these tests indicate that Fourier-transform wave-front reconstruction works in a real system. For both bright and dim stars, a Hudgin-geometry Fourier-transform method produced performance comparable to that of the Palomar Adaptive Optics least squares. The Fried-geometry method had a noticeable Strehl ratio performance degradation of 0.043 in the K band (165-nm rms wave-front error added in quadrature) on a dim star. PMID- 12779151 TI - Phase retrieval for high-numerical-aperture optical systems. AB - We describe a phase retrieval approach for intensity point-spread functions of high-numerical-aperture optical systems such as light microscopes. The method calculates a generalized pupil function defined on a spherical shell, using measured images at several defocus levels. The resultant pupil functionsreproduce measured point-source images significantly better than does an ideal imaging model. Availability of pupil function information will facilitate new approaches to aberration correction in such systems. PMID- 12779152 TI - Observation of vacuum-ultraviolet Ar2* radiation gain at 126 nm produced by an ultrashort high-intensity laser pulse propagating in a hollow fiber. AB - We observed a small-signal gain of Ar2* emission at 126 nm by use of a hollow fiber to guide the high-intensity laser propagation in high-pressure Ar. The small-signal gain coefficient was measured to be 0.05 cm(-1) at 126 nm. Kinetic analysis revealed that the electrons produced by the high-intensity laser through an optical-field-induced ionization process initiated the Ar2* production processes. The increase in the emission intensity was measured to be exp(2.5), with an increase in the fiber length. PMID- 12779153 TI - Production of radially or azimuthally polarized beams in solid-state lasers and the elimination of thermally induced birefringence effects. AB - Production and amplification of radially and azimuthally (tangentially) polarized laser beams are demonstrated. Based on the different focusing between radially and tangentially polarized light in thermally stressed isotropic laser rods, Nd:YAG laser oscillators were developed to produce low-loss stable oscillation in a single polarization. Pure radially polarized light at 70 W with M2 = 2 and on axis impure radially polarized light at 150 W with M2 = 2.5 were achieved. The radially polarized beams were then amplified while good beam quality and polarization purity were retained. Complete elimination of thermal-birefringence induced aberrations was demonstrated. This should allow much better beam quality from rod-based high-power lasers. PMID- 12779154 TI - Low-threshold quantum-cascade lasers at 3.5 THz (lambda = 85 microm). AB - Chirped-superlattice quantum-cascade lasers are reported that emit at lambda approximately 85 microm (3.6 THz), which is to the authors' knowledge the longest wavelength demonstrated so far with this technology. Collected peak output powers of 1.5 mW per facet were measured at liquid-helium temperature, and a maximum operating temperature of 45 K was reached. Record low-threshold-current densities of 95 and 115 A cm(-2) were observed in pulsed and continuous-wave operation, respectively. For the latter, output powers of a few hundred microwatts are estimated at low temperatures. PMID- 12779155 TI - Ultralow-jitter, 1550-nm mode-locked semiconductor laser synchronized to a visible optical frequency standard. AB - Using high-bandwidth feedback, we have synchronized the pulse train from a mode locked semiconductor laser to an external optical atomic clock signal and achieved what is to our knowledge the lowest timing jitter to date (22 fs, integrated from 1 Hz to 100 MHz) for such devices. The performance is limited by the intrinsic noise of the phase detector used for timing-jitter measurement. We expect such a highly stable device to play an important role in fiber-network based precise time/frequency distribution. PMID- 12779156 TI - Full-field optical coherence tomography by two-dimensional heterodyne detection with a pair of CCD cameras. AB - A two-dimensional heterodyne detection technique based on the frequency synchronous detection method [Jpn. J. Appl. Phys. 39, 1194 (2000)] is demonstrated for full-field optical coherence tomography. This technique, which employs a pair of CCD cameras to detect the in-phase and quadrature components of the heterodyne signal simultaneously, offers the advantage of phase-drift suppression in interferometric measurement. Horizontal cross-sectional images are acquired at the rate of 100 frames/s in a single longitudinal scan, with a depth interval of 6 microm, making the rapid reconstruction of three-dimensional images possible. PMID- 12779157 TI - Ultrasound detection through turbid media. AB - Optical coherence-domain reflectometry and laser-based ultrasound detection have been combined with the use of adaptive optics to detect ultrasound through turbid media. The dynamic hologram in a photorefractive quantum-well device performs as a coherence gate that eliminates multiply scattered background. Quadrature homodyne detection conditions are selected by the choice of center wavelength of the pulse spectrum, requiring no active stabilization or feedback. A depth resolution of 30 microm was achieved, with a pulse duration of nominally 120 fs for ultrasound detection through turbid media up to optical thicknesses of 11 mean free scattering lengths. PMID- 12779158 TI - Evolution of the mean instantaneous intensity in fibers affected by polarization mode dispersion. AB - It is shown that the mean instantaneous intensity (MII) of a pulse propagating in an optical fiber affected by polarization mode dispersion is related to the frequency autocorrelation of the fiber's Jones matrix through a Fourier transform. A simple derivation of the diffusion equation satisfied by the MII and the autocorrelation function of a Jones matrix is described. PMID- 12779159 TI - Helmholtz dark solitons. AB - A general dark-soliton solution of the Helmholtz equation (with defocusing Kerr nonlinearity) that has on- and off-axis, gray and black, paraxial and Helmholtz solitons as particular solutions, is reported. Modifications to soliton transverse velocity, width, phase period, and existence conditions are derived and explained in geometrical terms. Simulations verify analytical predictions and also demonstrate spontaneous formation of Helmholtz solitons and transparency of their interactions. PMID- 12779160 TI - Parametric vector solitons in tetragonal crystals. AB - We introduce novel types of spatial vector soliton that can be generated in anisotropic optical media, such as tetragonal crystals with third-order nonlinear susceptibility. We demonstrate that these vector solitons provide a nontrivial generalization to both conventional vector solitons of birefringent cubic media and parametric solitons supported by third-order cascaded nonlinearities. PMID- 12779161 TI - Electromagnetically induced transparency due to intervalence band coherence in a GaAs quantum well. AB - We demonstrate electromagnetically induced transparency in the transient optical response in a GaAs quantum well by using the nonradiative coherence between the heavy-hole and the light-hole valence bands. PMID- 12779162 TI - Interactions of discrete solitons with structural defects. AB - We investigated the interaction of discrete solitons with defect states fabricated in arrays of coupled waveguides. We achieved attractive and repulsive defects by decreasing and increasing, respectively, the spacing of one pair of waveguides in an otherwise uniform array. Linear and nonlinear propagation in the same samples show distinctly different properties. The role of the Peierls Nabarro potential in the interaction of the soliton with the defect is discussed. PMID- 12779163 TI - Demonstration of an all-optical switch by use of a multiwavelength mutual injection-locked laser diode. AB - Error-free all-optical packet switching is demonstrated by use of a multiwavelength mutual injection-locked Fabry-Perot laser diode. A 10-Gbit/s data signal is switched on and off with an extinction ratio of 16.9 dB when an optical control signal is turned off and on with a power difference of only 3 dB. PMID- 12779164 TI - Polarization-independent photochromic diffraction in a dye-doped liquid crystal. AB - We report the observation of polarization-independent photochromic diffraction in an azo-dye-doped liquid crystal. The generation of the phase grating is more than 90% independent of the polarization of the writing beams, and the diffraction by the phase grating is more than 90% independent of the polarization of the probe beam. Unpolarized lamp light was also used to generate real-time phase gratings and self-diffraction. For the first time to our knowledge, photochromic phase modulation and light diffraction that exhibit more than 90% polarization independence for both writing and probe beams were produced in an anisotropic liquid-crystalline material. PMID- 12779165 TI - Fabrication and performance of a medium-dependent SiO2/Si photonic heterostructure device. AB - An optical device based on a photonic bandgap heterostructure is designed, fabricated, and characterized. The sample contains two sets of Si/SiO2 photonic crystals with different periods. When the device is working in air, it reflects omnidirectionally both TE and TM mode lights at the wavelength near 1.3 microm. The reflectivity measured in the bandgap is higher than 98% in an incident angle range 0 degrees-70 degrees. When the device is surrounded with silica (n > 1.33), it permits the total transmission for the TM mode but prevents the TE mode from propagating, thus behaving as a polarization splitter. The experimental extinction ratio of the reflected TE/TM is approximately 31 dB. The uniformity of the device performance over a large sample area is demonstrated. PMID- 12779166 TI - Absorption losses in periodic arrays of thin metallic wires. AB - We analyze the transmission and reflection of electromagnetic waves calculated from transfer matrix simulations of periodic arrangements of thin metallic wires. The effective permittivity and the absorption of the arrangements of wires are determined. Their dependence on the wire thickness and the conductance of the metallic wires is studied. The cutoff frequency, or effective plasma frequency, is obtained and compared with analytical predictions. It is shown that the periodic arrangement of wires exhibits a frequency region in which the real part of the permittivity is negative while its imaginary part is very small. This behavior is seen for wires with thickness as small as 17 microm with a lattice constant of 3.33 mm. PMID- 12779167 TI - Relativistic scattering from moving fractally corrugated surfaces. AB - The solution to the problem of optical or electromagnetic-wave scattering from perfectly conducting rough surfaces modeled by a multiscale fractal function is generalized to the case of relative uniform translating motion with respect to the observer. The presence of motion can be treated by the relativistic frame hopping method by means of plane-wave simplification techniques, since the solution relevant to the static case is expressed in terms of a generalized expansion of Floquet modes. PMID- 12779168 TI - Intensity-related dynamics of femtosecond frequency combs. AB - We have performed systematic studies of intensity-related dynamics of the pulse repetition and carrier-envelope offset frequencies in mode-locked Ti:sapphire lasers. We compared the results far two laser systems that have different intracavity dispersion-compensation schemes. We found that the carrier-envelope phase noise and its dynamic response depend critically on the mode-locking conditions. An intensity-related shift of the laser spectrum was found to be instrumental in interpretations. PMID- 12779169 TI - Semianalytic treatment for propagation in finite photonic crystal waveguides. AB - We present a semianalytic theory for the properties of two-dimensional photonic crystal waveguides of finite length. For single-mode guides, the transmission spectrum and field intensity can be accurately described by a simple two parameter model. Analogies are drawn with Fabry-Perot interferometers, and generalized Fresnel coefficients for the interfaces are calculated. PMID- 12779170 TI - What is your diagnosis? Primary pulmonary tumour (carcinoma) with digital metastases. PMID- 12779171 TI - Safety of Cartrophen Vet in the dog: review of adverse reaction reports in the UK. AB - Suspected adverse reactions (SARs) reported for Cartrophen Vet (100 mg sodium pentosan polysulphate/ml) to the Veterinary Medicines Directorate in the UK for the period January 1991 to October 1999 were reviewed. Of the 161 reports, 28 were probably product related, 54 were possibly product related, 71 were unlikely to be related and eight were unclassified. An estimated real incidence of adverse reactions probably and possibly associated with Cartrophen Vet of 0.074 per cent on an individual dose basis was calculated (assuming only 10 per cent were documented due to underreporting). Sixty-two SARs (38.5 per cent) documented emesis, 22 (35.5 per cent) of which were product related (onset five to 15 minutes after administration). Sixty-eight SARs (42.2 per cent) documented general changes to demeanour, 10 (14.7 per cent) were product related (lethargy and/or mild depression and/or mild inappetence lasting up to two days after administration). Six SARs were considered likely to be associated with concurrently administered carprofen. Cartrophen Vet had a low incidence of side effects that were mild and transitory. PMID- 12779172 TI - Pulmonary oedema in Swedish hunting dogs. AB - A syndrome of acute dyspnoea after hunting in 16 Swedish hunting dogs is characterised. Radiographic pulmonary infiltrates interpreted as pulmonary oedema were found in the acute stage. In 12 dogs, the infiltrates regressed after five to 14 days. Subendocardial necrosis and pulmonary oedema were found at postmortem examination in four other dogs with acute and recurrent dyspnoea after hunting, and myocardial fibrosis in a further three dogs with a history of recurrent dyspnoea after hunting; none of these pathological changes was seen in dogs which had no previous history of dyspnoea after hunting. A pathogenetic mechanism is proposed whereby high catecholamine levels, present during hunting due to the stress of excitement and exercise, cause acute cardiac and pulmonary lesions in some susceptible dogs, similar to neurogenic or postictal pulmonary oedema. PMID- 12779173 TI - High-grade surface osteosarcoma in a dog. AB - A case of a high-grade surface osteosarcoma in the thoracic limb of a dog is reported. Radiographically, there was no bone involvement but there was increased soft tissue opacity. An aggressive osteosarcoma was diagnosed on the basis of early metastatic disease and histopathological examination. Previously reported surface osteosarcomas in the dog have been periosteal and parosteal osteosarcomas which are generally considered to have a low to intermediate biological activity. The tumour described here had radiographical, gross pathological, histopathological and biological features consistent with human high-grade surface osteosarcoma. The characteristics of surface osteosarcomas are briefly reviewed. PMID- 12779175 TI - Neuronal ceroid-lipofuscinosis and hydrocephalus in a chihuahua. AB - A two-year-old, female chihuahua presented with a six-month history of visual dysfunction. Computed tomography revealed dilation of the lateral ventricles in the central nervous system (CNS). The dog was tentatively diagnosed as having hydrocephalus and a month later was euthanased at the owner's request. The skull was expanded and dome-like in shape and an open fontanelle was observed on postmortem examination. Histologically, swollen neurons possessing yellowish pigment granules in the cytoplasm were observed throughout the CNS. These storage materials stained positively with periodic acid Schiff, Schmorl method for lipofuscin and oil red O for lipid, and showed autofluorescence under fluorescence microscopy. Ultrastructurally, the storage materials consisted of dense lamellar structures. This case was unique in having ceroid-lipofuscinosis in association with hydrocephalus. PMID- 12779174 TI - Hypercalcaemia associated with a retroperitoneal apocrine gland adenocarcinoma in a dog. AB - A seven-year-old, entire female golden retriever was presented with a history of polyuria/polydipsia and progressive dysuria. Clinical examination, radiography and ultrasonography demonstrated urinary retention due to a large soft tissue mass in the retroperitoneal space. Laboratory findings revealed paraneoplastic hypercalcaemia. Fine-needle aspiration cytology of the mass suggested an epithelial tumour, resembling an apocrine gland carcinoma of the anal sac. Following euthanasia and necropsy, the histopathological diagnosis of the retroperitoneal mass was apocrine gland adenocarcinoma. Despite ante- and postmortem examination, no perineal or anal sac tumour was found. The retroperitoneal tumour in this case could be a very large lymph node metastasis from an occult primary apocrine carcinoma of the anal sacs, or it could represent the first case of an ectopic apocrine gland carcinoma of the retroperitoneal space in a dog. PMID- 12779176 TI - Extrahepatic biliary tract surgery in the cat: a case series and review. AB - Four cases of extrahepatic biliary tract surgery in the cat are described. The causes of the disease were inflammation of the gallbladder, distal common bile duct (CBD) or major duodenal papilla, and traumatic avulsion of the CBD. Bile peritonitis was present in two of the cats. Biliary enterostomy was performed in three cats, two of which were euthanased at five weeks and three months postsurgery; the third was alive at the time of writing, four months postsurgery. Cholecystectomy was curative in one cat. A literature review reveals high early mortality following biliary diversion, with only 50 per cent of cases surviving more than two weeks, and 23 per cent surviving more than six months. Surviving cats had repeated intermittent vomiting and anorexia that responded to antibiotics. No postoperative mortality was seen when biliary diversion was avoided. Whenever biliary enterostomy or temporary diversion methods are performed, a poorer prognosis should be offered due to the increased likelihood of postoperative complications and mortality. PMID- 12779177 TI - Use of an ankle-foot orthosis in a dog with traumatic sciatic neuropathy. AB - Traumatic unilateral sciatic neuropathy secondary to a gunshot wound was diagnosed in a seven-month-old, intact male golden retriever. Findings on neurological assessment, performed after cardiovascular stabilisation, were incompatible with a return to normal limb function. An ankle-foot orthosis was used to correct excessive flexion of the digits and tarsus. This device greatly improved the dog's willingness to ambulate and could serve as an alternative to amputation in companion animals with severe sciatic axonotmesis or neurotmesis. Complications associated with chronic tibiotarsal hyperflexion necessitated a pantarsal arthrodesis one year after initial presentation; the dog also developed cranial cruciate disease in the affected limb three years after the gunshot injury. PMID- 12779178 TI - Progress under the BVA and KC hip dysplasia scheme. PMID- 12779179 TI - Focusing on issues of the day at Congress 2003. PMID- 12779180 TI - Professional guidance on handling animal and human abuse. PMID- 12779181 TI - Towards excellence in orthopaedics. PMID- 12779182 TI - Treatments for depression: wisdom imparted from treatments discarded. AB - Many different forms of treatment have been used with patients suffering from depression. It can be difficult to examine current treatments and anticipate problems, mistakes, and limitations. However, a historical perspective allows an appreciation of the strengths and limitations of older treatments, and sheds light on current strategies. A variety of effective treatments have been discarded and forgotten. Important lessons can be learned by reviewing the history of treatments for depression. Many treatment strategies were based on accidental discovery, but nonetheless were found effective in reducing the symptoms of depression. Both case examples and group studies have documented the effectiveness of these older treatments. A review of discarded treatments reveals how a biological model can be limited in its ability to produce a lasting cure. Caution and skepticism are encouraged when innovative treatments are proposed. PMID- 12779183 TI - Fluoxetine's effects on cognitive performance in patients with traumatic brain injury. AB - OBJECTIVE: There are preclinical data showing that fluoxetine stimulated expression of Brain Derived Neurotrophic Factor (BDNF) and its specific tyrosine kinase receptor, and caused neuritic elongation and increased dendritic branching density of CA3 hippocampal pyramidal cell neurons in rodents. The latter effect of fluoxetine has been referred to as neuronal remodeling. In view of this preclinical data, we wondered if specific cognitive measures could serve as novel therapeutic targets for fluoxetine in head-injured patients. Theoretically, fluoxetine-induced "neuronal remodeling" might improve cognition, independently of a primary effect on mood. METHOD: In an open-label pilot investigation, fluoxetine hydrochloride (Prozac; 20-60 mg/day) was administered to a heterogeneous group of five head-injured patients with either no or moderate depression for a period of eight months. These patients had no histories of prior treatment with antidepressant medications. They were administered cognitive and memory tests at baseline and after eight months of treatment on fluoxetine. RESULTS: The preliminary results showed that fluoxetine improved mood, in addition to improving performance on the Trail Making Test Part A, an attentional motor speed task, and the letter-number sequencing subtest of the WAIS-III, a measure reflecting "working memory." CONCLUSIONS: Although fluoxetine had beneficial effects on some measures of cognition, more work is needed to connect these improvements with neuronal remodeling. PMID- 12779184 TI - Expression of anger and alexithymia in patients with psychogenic excoriation: a preliminary report. AB - OBJECTIVE: Psychogenic excoriation (PE), which is characterized by lesions formed by self-picking, has a significant place among the dermatoses related to psychological factors. Emotions, particularly anger that cannot be expressed, may be important in the etiology. The objective of this study was to evaluate the sociodemographic characteristics of patients with PE and with another psychodermatosis, and compare them in terms of anger, manner of anger expression, and alexithymia. METHODS: Thirty-one consecutive subjects with PE and thirty-one patients with chronic urticaria were recruited from an outpatient dermatology clinic. All of the subjects completed Toronto Alexithymia Scale and Trait Anger and Anger Expression Scale. RESULTS: PE patients had higher levels of anger (p < .01), tended not to show their anger (p < .05), and were more alexithymic (p < .05). There was also a positive correlation between anger and alexithymia scores (r = .49, p < .05). DISCUSSION: PE, a severe and chronic psychiatric and dermatological problem, may be related to affect-regulation, particularly anger and alexithymia. Due to the fact that it has a different place among psychodermatoses, individuals with PE might benefit from learning how to regulate their affects other than by excoriation. PMID- 12779185 TI - Psychogenic cough: a profile of 32 cases. AB - OBJECTIVE: Psychogenic or nervous cough has generally been viewed as a clinical oddity and largely ignored. Though it is not rare, its clinical profile is not yet well known. METHOD: In the present study, a series of 32 cases of psychogenic cough is reported. RESULTS: Of 32 cases, there were 19 females (59.4 percent) and 13 males (40.6 percent). A majority of patients had cough for more than one month. School phobia (in children) and fear of rejection and need for attention were the most common precipitating or perpetuating factors. Twenty patients (62.5 percent) had a psychiatric disorder, the most common being conversion disorder (21.9 percent) followed by mixed anxiety and depressive disorder (12.5 percent). Drug therapy and/or psychotherapy were used as treatment with 12 cases (37.5 percent) showing remission, 16 cases (50.0 percent) having improvement, and 4 cases (12.5 percent) continuing with the complaint. CONCLUSION: Further studies are warranted to study the treatment and outcome of this important psychiatric disorder. PMID- 12779186 TI - Acute stress reactions to recent life events among women and men living with HIV/AIDS. AB - OBJECTIVE: This study examined the prevalence of acute stress reactions to recent life events among persons living with HIV/AIDS. A second aim was to investigate the relationship of acute stress reactions among HIV-infected men and women to posttraumatic stress disorder (PTSD) symptoms to previous traumatic life events. METHOD: Participants included 64 HIV-seropositive persons (33 men and 31 women) drawn from a larger study examining the effects of group therapy on quality of life and health behavior. These individuals were assessed at baseline on demographic and medical status characteristics and (PTSD) symptoms andthen randomly assigned to either receive group therapy plus education or education alone. Three months later they were assessed for acute stress reactions to recent life events. RESULTS: Nearly a third (31.3 percent) of the participants reported levels of acute stress reactions to recent life events that met all symptom criteria for the diagnosis of acute stress disorder. However, only 9.4 percent of the respondents described a recent stressful life event that was threatening to the life or physical integrity of themselves or others. Acute stress reactions to recent life events were significantly and positively related to experiencing PTSD symptoms to prior traumatic life events. Acute stress did not differ significantly by gender, AIDS status, or whether or not participants had received 12 weeks of group therapy. CONCLUSIONS: A subset of individuals with HIV/AIDS experience high levels of acute stress reactivity to life events considered non traumatic. HIV-infected individuals who react strongly to ongoing life stressors are more likely to have developed PTSD symptoms in response to previous traumatic life events. PMID- 12779187 TI - Is depressed affect a function of one's relationship with God?: Findings from a study of primary care patients. AB - OBJECTIVE: This study examines the association between a self-reported loving relationship with God and the presence of depressed affect. Building on prior clinical and epidemiologic research on religious factors in mental health, it seeks to extend consideration to internal religious resources. METHOD: Data are from 205 primary care outpatients who completed a self-administered survey inquiring about their relationship with God, their mental and physical health, and various religious and psychosocial issues. The principal dependent construct is the depressed affect subscale of the General Well-Being Scale. The principal independent construct is a validated eight-item self-report measure of loving and being loved by God based on a theoretical taxonomy developed by Sorokin. RESULTS: Hierarchical ordinary least squares regression was used to investigate the association between this construct, which Sorokin termed "religious love," and the measure of depressed affect. After controlling for sets of hypothesized mediating factors (multi-item measures of religious involvement, social resources, psychological resources, and physical health status) in five successive models, as well as several key sociodemographic variables, the statistically significant inverse association between these two constructs in the baseline model (beta = -.29, p < .01) remained strong and statistically significant (beta = -.21, p < .05). CONCLUSIONS: These findings raise the possibility that a loving relationship with God may exert a protective effect on psychological distress. One's relationship with God thus may represent an important personal resource for mitigating the emotional consequences of poor health and other deleterious life circumstances, as well as marker for successful religious coping. PMID- 12779188 TI - Fire-settng behavior associated with Klinefelter syndrome. AB - This is a report on a male patient with a previous history of psychotic disorder who was arrested for arson. During the following hospitalization in a psychiatric unit, a genetic syndrome, namely Klinefelter syndrome (47,XXY), was diagnosed. The association between firesetting behavior and Klinefelter syndrome is reviewed, and the underlying explanatory hypothesis is discussed. PMID- 12779189 TI - Neurosarcoidosis presenting as psychosis and dementia: a case report. AB - Neurosarcoidosis is a rare disorder in which psychosis and dementia may occur. They usually appear subsequently to the diagnosis of pulmonary sarcoidosis. We report on a 39-year-old patient who presented with long-term decline and acute onset of psychosis and delirium, and who was found to have neurosarcoidosis. PMID- 12779190 TI - Modafinil augmentation of mirtazapine in a failure-to-thrive geriatric inpatient. AB - The failure-to-thrive syndrome in geriatric patients is marked by social withdrawal, apathy, depression, anorexia, and cognitive impairment. For therapy to be effective, the treatment plan must target several of theseareas. This case report describes one such course of treatment for a patient with multiple myeloma with failure-to-thrive who was successfully treated with modafinil and mirtazapine. By using combination pharmacotherapy, we were able to achieve immediate results in a gravely ill patient. PMID- 12779191 TI - Lessons to be learnt from the International Subarachnoid Haemorrhage Trial (ISAT). PMID- 12779192 TI - Factors likely to affect the location and provision of future neurosurgical services in the UK. PMID- 12779193 TI - Shaken baby syndrome. PMID- 12779194 TI - Shaken baby syndrome. PMID- 12779195 TI - Shaken baby syndrome. PMID- 12779196 TI - Shaken baby syndrome. PMID- 12779197 TI - Diagnostic criteria for the clinical syndrome of internal disc disruption: are they reliable? AB - A new disease, named internal disc disruption (IDD), has provoked debate. Some insist that discography is specific for the diagnosis, while others disagree. Without scientific verification, some doctors have performed invasive operations for this uncertain disease. It is necessary to explore the diagnostic criteria and characteristics of IDD. We investigated the background, history, diagnostic methods and criteria of IDD by a review of the literature. The criteria for diagnosis of IDD are diverse. The minimum requirements for the diagnosis were the pattern of pain and the shape on discography. Although the pain pattern is important for the correct diagnosis, it depends on the subjective report of the patient. The diagnosis is up to the patient, and the examiner alone cannot make it. We conclude that IDD is not a real, but a hypothetical disease. Until scientific verification is forthcoming any invasive procedures should be restricted. PMID- 12779198 TI - To determine the effect of metoclopramide on gastric emptying in severe head injuries: a prospective, randomized, controlled clinical trial. AB - To determine the effect of 8-hourly administration of 10 mg intravenous metoclopramide, over a 48-h period on gastric emptying in severe head injury (SHI), 22 patients were prospectively randomized (Glasgow Coma Score of 3-8) to receive 2 ml of intravenous metoclopramide or 2 ml of 5% saline 8-hourly for 48 h. Baseline and serial blood paracetamol absorption assays were performed at time (t) = 0, 15, 30, 45, 60, 90 and 120 min on day 0 and day 2. The area under the curve between the day 0 and day 2 was used to measure the degree of gastric emptying. In SHI, sequential doses of metoclopramide did not appear to improve gastric motility within subject comparisons (p = 0.65) and between subject comparisons (placebo p = 0.4 and drug p = 0.12). Metoclopramide has no significant prokinetic effect on gastric emptying in SHI patients when given in the early postinjury period. PMID- 12779199 TI - Traumatic brain injury in childhood: intensive care time series data and outcome. AB - Age-specific norms are necessary to determine potential secondary brain insult after head injury in children. We describe and quantify the secondary physiological derangement recorded in children of different ages following traumatic brain injury, and relate it to outcome at 12 months post-injury. Prospective time-series data (including intracranial pressure, arterial blood pressure, cerebral perfusion pressure, oxygen saturation, temperature and heart rate) downloaded from ICU monitors, were examined to identify abnormal (i.e. outside normal age-specific limits) recordings lasting more than 5 min. Cumulated total duration of derangement was calculated for each parameter and as a percentage of the time that the ICP monitor was in situ. Univariate and multivariate logistic regression modelling was used to evaluate predictors of outcome. Age-specificity allows realistic comparisons of physiological data among children. Duration of age-specific derangement of CPP was found to predict outcome (dead v. alive: p = 0.003 and Glasgow Outcome Score 1-3 v. 4-5, i.e. poor v. independent outcome p = 0.004). PMID- 12779200 TI - Costs of thalamic deep brain stimulation for movement disorders in patients with multiple sclerosis. AB - Several studies have shown that thalamic deep brain stimulation (DBS) reduces tremor and improves hand performance in patients with multiple sclerosis (MS). The purpose of this paper is to describe the cost implications of DBS in MS patients and to highlight postoperative medical requirements that can be associated with this therapy. In a prospective study of thalamic DBS in MS patients the mean equipment costs were pounds 4769 (median pounds 7010, Medtronic, 1998 prices); mean neurosurgical inpatient costs per operated patient (n = 15) were pounds 4848 (range pounds 1982-8920, median pounds 5110); and mean in-patient postoperative rehabilitation cost pounds 4602 (range pounds 0-32,225, median pounds 1783). In addition there were transport and follow up costs. Mean neurosurgical inpatient stay following stereotactic DBS implantation was 15 days (median 12 days); and mean inpatient, postoperative rehabilitation stay 54 days (median 25 days). Although there were significant improvements in hand function and tremor reduction at 12 months postoperation, the level of patient performance in activities of daily living, their perception of their handicap and ipse facto the amount of home support required were unchanged from preoperative levels. This study has highlighted significant unforeseen medical requirements and costs that can occur in MS patients who have thalamic DBS surgery. PMID- 12779201 TI - Therapeutic decision and management of aneurysmal subarachnoid haemorrhage based on computed tomographic angiography. AB - The purpose of this study was to evaluate the potential of high quality computed tomographic angiography (CTA) to replace digital subtraction angiography (DSA) in cases of ruptured saccular aneurysms and perform early surgical clipping or coiling on the basis of CTA alone. In a prospective study, 100 patients with aneurysmal subarachnoid haemorrhage (SAH) diagnosed by computed tomography underwent CTA. CTA revealed a total of 118 aneurysms including all ruptured aneurysms. A decision of direct surgical clipping, endovascular coiling or therapeutic abstention was made in 89 cases (89%) on the basis of CTA alone. Sixty-one direct surgical procedures were performed after CTA. Twenty-six cases underwent DSA for immediate endovascular treatment of the ruptured aneurysm. In 11 cases (11%), a DSA was performed prior to the therapeutic decision because of unclear aneurysm. Four cases were not treated because of initial poor clinical grade. The surgical findings were compared with CTA data and were considered accurate in all but one case. All patients underwent postoperative DSA within 10 days after SAH. The sensitivity and the specificity of CTA for the detection of all aneurysms, as compared with postoperative DSA, were 95.1 and 100%, respectively. A total of six unruptured aneurysms were missed initially, but were visible retrospectively on CTA in all but one case and were found in patients with multiple aneurysms in whom the ruptured aneurysm was detected by CTA. Current quality CTA allows reliable pretreatment planning for the majority of cases of aneurysmal subarachnoid haemorrhage and diminishes the pretreatment evaluation time critically. Complementary pretreatment DSA is required in situations where CTA characteristics of the ruptured aneurysm is unsatisfactory. PMID- 12779202 TI - Assessment of fusion after anterior cervical discectomy. AB - There is no established method to assess fusion in patients following anterior cervical discectomy. In this study we have made a series of measurements to detect movement between vertebrae adjacent to an operated space. The absence of movement implies structural union between adjacent vertebrae. Measurements have been made in two distinct surgical groups. Group A patients had anterior cervical discectomy with insertion of a BOP graft into the disc space. Group B patients underwent simple anterior cervical discectomy with no spacer or graft material inserted, the disc space being left empty. Details of the measurements and interpretation of results are described. In the absence of a 'gold standard' to assess bony union we propose that these measurement methods provide an objective and scientific method to assess fusion at the operated level after anterior cervical discectomy. Objective measurement of fusion will allow comparison between different surgical techniques that claim fusion as an end point. It will also become possible to study the influence of fusion on clinical outcome in different surgical populations. PMID- 12779203 TI - How accurate is magnetic resonance angiography in predicting neurovascular compression in patients with trigeminal neuralgia? A prospective, single-blinded comparative study. AB - Ninety-two patients with trigeminal neuralgia (TGN) were investigated prior to posterior fossa surgery with magnetic resonance imaging (MRI) and contrast enhanced magnetic resonance angiography (MRA). The preoperative investigation was matched to one consultant neuroradiologist (co-author) who was blinded to the side of symptomotology. The imaging results were compared with the operative findings in all patients. In 76 patients MRA showed present neurovascular compression in accordance with surgical findings. Eight cases had no compression either on MRA or intraoperatively. Eight predictions of no compression were false and there was no false positive. Based on surgical findings, the sensitivity of MRA was 90.5% and the specificity 100%. In 19 cases MRA predicted bilateral compression of the trigeminal nerves. Only two cases had clinically bilateral TGN. We conclude that MRA with gadolinium enhancement is an extremely sensitive and specific method for demonstrating compression in TGN. As a result posterior fossa surgery can be recommended with confidence, and microvascular decompression remains the treatment of choice for TGN at the authors' centre. PMID- 12779204 TI - Pericranial flap fixation using titanium miniplates and screws. AB - Opening the frontal sinuses is a common occurrence during neurosurgical procedures involving the anterior cranial base. Obliteration of the exposed sinuses is usually accomplished packing them with various materials and/or covering them with an anteriorly-based pericranial flap. In the absence of more extensive cranial base defects, the flap only needs to be wrapped around the sinus and secured against its posterior wall. We describe the technique for and the results of using titanium miniplates and screws to secure the pericranial flap to the inner surface of the frontal bone. In 10 patients treated with this alternative technique, the pericranial flap provided an excellent separation between the sinuses and the intradural compartment. No cases of cerebrospinal fluid leakage or infections were observed. PMID- 12779205 TI - The pretzel sign: angiographic pattern of tortuous intra-aneurysmal blood flow in a giant serpentine aneurysm. AB - Giant serpentine aneurysms (GSAs) form a specific subgroup of giant cerebral aneurysms that have pathognomonic angiographic features. We report the angiographic findings of a GSA demonstrating a striking convoluted dynamic flow pattern, which we have called the 'pretzel sign'. The aneurysm was successfully treated by permanent occlusion of the parent vessel using a detachable balloon. GSAs should be identified prior to treatment in view of their particular management requirements. PMID- 12779206 TI - Spinal pneumorrhachis. AB - The term pneumorrhachis refers to the presence of air in the spinal canal. There are only a few previously reported cases in the literature. A case of air within the cervical and lumbar subarachnoid spaces, and a second case of air within the cervical subarachnoid space due to traumatic pneumocephalus are presented in this study. PMID- 12779207 TI - Remote hypertensive intracerebral haematoma following clipping of an intracranial aneurysm. AB - We report a case of a 58-year-old woman who presented with an aneurysmal subarachnoid haemorrhage. Immediately following clipping of this aneurysm, she had a spontaneous hypertensive bleed in the contralateral hemisphere. Although very unusual, hypertensive episodes following aneurysmal subarachnoid haemorrhage must carry a risk of such an intracranial event. PMID- 12779208 TI - Hindbrain hernia masquerading as postpartum subarachnoid haemorrhage. AB - A 29-year-old postpartum lady presented with a headache suggestive of subarachnoid haemorrhage. Investigations were negative for sources of haemorrhage, but revealed a hindbrain hernia. Hindbrain hernia should be considered as a cause of headache postpartum, as repeated Valsalva manoeuvres performed during vaginal delivery may further aggravate tonsillar decent. PMID- 12779209 TI - Pseudotumour cerebri, CSF rhinorrhoea and the role of venous sinus stenting in treatment. AB - We present a case of pseudotumour cerebri (PTC), which is important as it illustrates the effects of chronically raised CSF pressure, the relationship between PTC and venous sinus obstruction and the successful treatment of PTC using a venous sinus stent. A 38-year-old woman, previously diagnosed with PTC and unsuccessfully treated 10 years previously re-presented with spontaneous CSF rhinorrhoea. Imaging revealed dramatic changes of chronically raised CSF pressure and a defect in the anterior cranial fossa. The CSF leak was corrected surgically and a lumbo-peritoneal shunt inserted to correct a large postoperative subgaleal CSF collection. Direct retrograde cerebral venography (DRCV) demonstrated venous sinus obstruction due to a filling defect. This was associated with a pressure gradient and a high superior sagittal sinus pressure. The venous sinus obstruction was successfully treated with a venous sinus stent and the lumbo peritoneal shunt was removed. Chronically raised CSF pressure in untreated cases of PTC may cause widespread changes in the skull, which in this case, culminated in a spontaneous CSF leak despite relatively mild headache and visual symptoms. Furthermore, cases of PTC secondary to venous sinus obstruction may be successfully treated using venous sinus stenting. The index of suspicion for venous sinus stenosis or obstruction should be raised in the investigation of patients with PTC. PMID- 12779210 TI - Nasal polyps: not always an ENT problem. PMID- 12779211 TI - Britain's ailing health service: a question of money or a question of morale? PMID- 12779212 TI - Contralateral headache. PMID- 12779213 TI - Resolution of papilloedema after neurosurgical decompression for primary Chiari I malformation. PMID- 12779214 TI - The advantages of frameless stereotactic biopsy over frame-based biopsy. PMID- 12779215 TI - Axial and radial diffusion coefficients in a liquid chromatography column and bed heterogeneity. AB - The axial and transverse diffusion coefficients of a band of iodine in a chromatographic column were measured optically as a function of time. It was found that the axial diffusion coefficient remains constant even when the edges of the sample band get close to the wall. By contrast, the radial diffusion coefficient decreases progressively with increasing time when the edges of the sample band leave the core region and begin to diffuse inside the wall region. The local axial and transverse diffusion coefficients of the band decrease from the column center toward the wall. Hence, the increase in local height equivalent to a theoretical plate observed in the region close to the wall must be explained by increasing mass transfer resistances and degree of heterogeneity of the bed. PMID- 12779216 TI - Chromatographic classification and comparison of commercially available reversed phase liquid chromatographic columns using principal component analysis. AB - A total of 135 commercially available alkyl, cyano, phenyl, perfluorinated, polar embedded, enhanced polar selectivity (i.e., polar/hydrophilic endcapped), "Aqua type" and a variety of novel phases including some non-silica based stationary phases have been characterised in terms of their surface coverage, hydrophobic selectivity, shape selectivity, hydrogen bonding capacity and ion-exchange capacity at pH 2.7 and 7.6. Principal component analysis has been used to provide a simple graphical comparison of the differences/similarities between columns in the entire database and differing subsets such as "Aqua type"/enhanced polar selectivity phases. The PCA has been correlated to the phase's ability to analyse a range of hydrophilic bases. PMID- 12779217 TI - Preparative separation and purification of squalene from the microalga Thraustochytrium ATCC 26185 by high-speed counter-current chromatography. AB - High-speed counter-current chromatography (HSCCC) was successfully applied to the preparative separation and purification of squalene from microalgae. Crude squalene was obtained from the microalga Thraustochytrium ATCC 26185 by extraction with organic solvents. The crude squalene was further separated using a waterless two-phase solvent system composed of n-hexane-methanol (2:1, v/v). The upper phase as the mobile phase was pumped into the column at a flow-rate of 2.0 ml min(-1) in the tail-to-head elution mode. The fractions purified and collected were analyzed by high-performance liquid chromatography. The method yielded 0.2 mg squalene at 96% purity from 150 mg of the crude squalene (0.14% squalene) with 95% recovery. The separation of squalene by HSCCC was completed in 90 min. PMID- 12779218 TI - Liquid chromatography-electrospray ionisation-mass spectrometry based method for the simultaneous determination of algal and cyanobacterial toxins in phytoplankton from marine waters and lakes followed by tentative structural elucidation of microcystins. AB - A liquid chromatography (LC)-based method with mass spectrometric (MS) detection was developed for simultaneous determination of various algal and cyanobacterial toxins extracted from phytoplankton occurring world-wide in marine waters and lakes. The method enables quantification of saxitoxin, anatoxin-A, domoic acid, nodularin, microcystins, okadaic acid and dinophysistoxin-1 with a single chromatographic run. In addition, the applied chromatographic conditions allow isolation and identification of substances suspected to be "new" microcystins (cyclic peptides) by fraction collection, hydrolysis, derivatisation of resulting free amino acids with the modified chiral Marfey's reagent N-alpha-(2,4-dinitro-5 fluorophenyl)-L-valinamide (L-FDVA) and enantioselective analysis of the amino acid derivatives by LC-ESI-MS. PMID- 12779219 TI - Separation and determination of denatured alpha(s1)-, alpha(s2)-, beta- and kappa caseins by hydrophobic interaction chromatography in cows', ewes' and goats' milk, milk mixtures and cheeses. AB - Caseins alpha(s1)-, alpha(s2)-, beta- and kappa- from raw cows', ewes' and goats' milk were separated and determined by hydrophobic interaction chromatography (HIC) by using a Propyl column (Eichrom) in the presence of 8.0 M urea in the mobile phase. The method is based on fast and easy solubilization of real raw samples by 4.0 M guanidine thiocyanate followed by the HIC analysis, without any preliminary precipitation or separation of the casein fraction. Elution conditions have been optimized by analyzing commercial single bovine standard caseins and their mixture. In the optimized chromatographic conditions the four casein fractions were separated in less than 45 min. A linear relationship between the concentration of casein and peak area (UV absorbance detector at 280 nm) has been obtained over the concentration range of 0.5 to 40 microM. The detection limit for alpha-, beta- and kappa-caseins ranged between 0.35 and 0.70 microM. The precision of the method was evaluated, the coefficient of variation for alpha-, beta- and kappa-casein determination ranging between 3.0 and 6.0%. The method has been validated by the analysis of reference skim milk powder (BCR 063R) certificated for total nitrogen content. The method was applied to commercial casein mixture and to the qualitative and quantitative analysis of casein fractions in unprocessed, raw cows', goats' and ewes' milk (10 samples analyzed for each species), in one sample of unprocessed buffalos' milk and in commercial cheeses (mozzarella, robiola, ricotta and stracchino). Binary mixtures of milk (cow/goat and cow/ewe) were also analyzed and the ratio between casein peak areas (alpha(s1)/kappa, alpha(s2)/beta, beta/kappa and alpha(s2)/alpha(s1)) of the HIC chromatograms was proposed and discussed in order to evaluate a possible application of this method to detect milk adulteration. PMID- 12779220 TI - Determination of the antibiotic chloramphenicol in meat and seafood products by liquid chromatography-electrospray ionization tandem mass spectrometry. AB - A confirmatory method based on isotope dilution liquid chromatography electrospray ionization tandem mass spectrometry is described for the determination of the antibiotic chloramphenicol (CAP) in foods. The method is quantitative and entails liquid-liquid extraction followed by a clean-up step on a silica gel solid-phase extraction cartridge. Mass spectral acquisition is done in the negative ion mode applying multiple reaction monitoring of two diagnostic transition reactions for CAP (m/z 321 --> 257 and m/z 321--> 152). In addition, the presence of two chlorine atoms in the CAP molecule provides further analyte certainty by assessing the 37Cl/35Cl ratio using the transition reactions m/z 323 --> 257 and m/z 323 --> 152. Validation of the method in chicken meat is conducted according to the latest European Union criteria for the analysis of veterinary drug residues at levels of 0.05, 0.10, and 0.20 microg/kg, employing [2H5]-chloramphenicol as internal standard. The decision limit and the detection capability were calculated at 0.01 microg/kg and 0.02 microg/kg, respectively. At the lowest fortification level (i.e. 0.05 microg/kg), precision values below 14 and 17% were achieved under repeatability and within-laboratory reproducibility conditions, respectively. The accuracy of the method was within 20, 15, and 5% of the target values at the 0.05, 0.10, and 0.20 microg/kg fortification levels, respectively. The applicability of this procedure was demonstrated by the analysis of other meat (turkey, pork, beef) and seafood (fish, shrimps) products. The method is robust and suitable for routine quality control operations, and more than 200 sample injections were performed without excessive pollution of the mass spectrometer or loss of LC column performance. PMID- 12779221 TI - Rapid separation of chlorophylls a and b and their demetallated and dephytylated derivatives using a monolithic silica C18 column and a pyridine-containing mobile phase. AB - A monolithic C18-bonded silica rod column (Merck Chromolith) was applied to the separation of mixtures of chlorophylls a and b and their derivatives originated by hydrolysis of the phytyl ester linkage (chlorophyllides), loss of the central Mg atom (pheophytins), or both processes (pheophorbides). Mobile phases containing two different ion-pair reagents, ammonium acetate and buffered pyridine, were tested. Both eluents achieved the resolution of the eight pigments in less than 5 min. The method based on the pyridine-containing mobile phase was applied to the separation of chlorophylls and their green coloured degradation products in senescing leaves of deciduous trees, green fruits and a marine microalga. PMID- 12779222 TI - Rapid identification and quantitation of compounds with forensic interest using fast liquid chromatography-ion trap mass spectrometry and library searching. AB - A fast liquid chromatography-electrospray tandem mass spectrometric (LC-ESI-MS MS) method by using a monolithic column, gradient elution and ion trap mass spectrometer was developed for 14 forensically interesting and chemically different compounds. All compounds were eluted within 2.5 min and the total analysis time was 5 min including stabilisation time required for the next injection. All the compounds, basics, neutrals and acids were efficiently ionised by positive ion ESI. A laboratory library including MS-MS spectra and retention times was developed and tested. Results with 476 standard samples and 50 authentic samples showed that the compounds studied can be unambiguously identified with the library. A quantitative method was developed for the compounds using external calibration. The evaluation process showed good linearity of the method and reasonable repeatability. Limits of detection ranged from 10.0 to 50.0 ng/ml. PMID- 12779223 TI - Prediction of the plate height of capillary columns operated at any inlet pressure of the carrier gas by using few retention data measured under isobaric conditions. AB - Programming inlet pressure in gas chromatography permits to decrease the analysis time without changing the elution order of compounds of different polarity whose relative retention changes with changing temperature. The choice of the best values of the inlet pressure and flow-rate of the carrier gas often requires many preliminary analyses with different parameters to be carried out. A method for the prediction of the separation by starting from few experimental data measured in isothermal and isobaric conditions decreases the time required for the optimisation of the analysis. The efficiency of the separation depends on the change of the theoretical plate height at various pressures and temperatures, due to pressure drop along the column. By calculation of the diffusion coefficients of the analysed compounds into the mobile and stationary phase it is possible to evaluate the column efficiency and predict the number of theoretical plates at any inlet pressure. A procedure for the prediction of the plate height of a capillary column at any inlet pressure of the carrier gas and column temperature by using retention data of polar and non-polar compounds (1-alcohols and linear alkanes) obtained in few isobaric runs is described. PMID- 12779224 TI - Mixed chiral stationary phase containing modified resorcinarene and beta cyclodextrin selectors bonded to a polysiloxane for enantioselective gas chromatography. AB - Both a resorcinarene with pendant L-valine diamide groups (used as hydrogen bonding selector) and a permethylated beta-cyclodextrin (used as inclusion-type selector) were chemically bonded to poly(hydromethyl)dimethylsiloxane in a one pot reaction via Pt-catalyzed alkene hydrosilylation. This novel mixed chiral stationary phase (mixCSP) named Chirasil-Calixval-Dex resembles a combination of the known chiral stationary phases (CSPs) Chirasil-Calixval and Chirasil-Dex and it was used successfully in enantioselective gas chromatography toward a unified enantioselective GC separation system. It is demonstrated that Chirasil-Calixval Dex retains the individual enantioselectivites of the single components. Thus the enantiomers of apolar hydrocarbons as well as polar amino acid derivatives can be separated with the mixed CSP. PMID- 12779225 TI - Identification and confirmation of traces of chlorinated fatty acids in fish downstream of bleached kraft pulp mills by gas chromatography with halogen specific detection. AB - Methyl esters of threo-9,10-dichlorooctadecanoic, threo-7,8-dichlorohexadecanoic, and threo-5,6-dichlorotetradecanoic acids, present in transesterified extracts of filets, gonad, intestinal fat and carcass of white sucker (Catostomus commersoni) sampled in receiving waters of bleached kraft pulp mill effluents, were identified by gas chromatography with halogen-specific detection (XSD). Identification was based on (1) a comparison of the retention times of a sample peak with a prospective reference standard on two stationary phases of very different polarities by spiking, and (2) elution behavior of configurational and positional isomers of dichloro fatty acid methyl esters. PMID- 12779226 TI - Automated fast extraction of nitrated polycyclic aromatic hydrocarbons from soil by focused microwave-assisted Soxhlet extraction prior to gas chromatography- electron-capture detection. AB - An approach for the automated fast extraction of nitrated polycyclic aromatic hydrocarbons (nitroPAHs) from soil, using a focused microwave-assisted Soxhlet extractor, is proposed. The main factors affecting the extraction efficiency (namely: irradiation power, irradiation time, number of cycles and extractant volume) were optimised by using experimental design methodology. The reduction of the nitro-PAHs to amino-PAHs and the derivatisation of the reduced analytes with heptafluorobutyric anhydride was mandatory prior to the separation-determination step by gas chromatography--electron-capture detection. The proposed approach has allowed the extraction of these pollutants from spiked and "real" contaminated soils with extraction efficiencies similar to those provided by the US Environmental Protection Agency methods 3540-8091, but with a drastic reduction in both the extraction time and sample handling, and using less organic solvent, as 75-85% of it was recycled. PMID- 12779227 TI - Screening and determination of pesticides in soil using continuous subcritical water extraction and gas chromatography-mass spectrometry. AB - In the present work the efficiency of water under subcritical conditions for the extraction of pesticides having a broad spectrum of polarities from soils was evaluated. The pesticides under study were carbofuran, hexachlorobenzene, dimethoate, simazine, atrazine, lindane, diazinon, methylparathion, alachlor, aldrin-R, metholachlor, chlorpyrifos, heptachlor epoxide, dieldrin, endrin, 4,4 DDT and metoxichlor. Optimization studies were carried out using a blank soil (Non-Polluted Soil 1, CLN-1, RTC) and a real soil which were previously spiked with the pesticide mixture and aged for 60 days. A laboratory-made aluminum oven with controlled temperature was used to carry out the leaching process with subcritical water, where it is placed a pre-heater and the extraction cell. The following variables were studied, keeping the pressure controlled about 1200 p.s.i.: the extraction temperature, the time of static and dynamic extraction and the flow-rate of water (1 p.s.i. = 6894.76 Pa). The extraction efficiency of the pesticides increases with the temperature trending to the quantitative extraction at temperatures near to 300 degrees C. After the extraction process, the analytes were transferred quantitatively to 5 ml dichloromethane, before the determination by GC-MS. The results indicate that under the optimized conditions mostly of the analytes are extracted quantitatively in 90 min with recoveries quite similar to those obtained by the standard Soxhlet extraction procedure. Alternatively, by using an extraction time of 25 min, the method can be used as screening for all the pesticides, with recoveries depending on their polarity. PMID- 12779228 TI - Attempt to unravel the composition of toxaphene by comprehensive two-dimensional gas chromatography with selective detection. AB - Comprehensive two-dimensional gas chromatography (GC x GC) coupled with micro electron-capture and time-of-flight mass spectrometric (TOF-MS) detection has been used to analyse technical toxaphene. An HP-1 x HT-8 column combination yielded highly structured chromatograms and revealed a complex mixture of over 1000 compounds what is significantly higher number than in any study before. The analysis of a mixture of 23 individual congeners and TOF-MS evaluation of technical toxaphene showed that the chromatogram is structured according to the number of chlorine substituents in a molecule. The nature of the compounds (bornane and camphene) does not appear to have any influence. The sum of the peak areas of all congeners in each group was calculated using laboratory-written software; based on these results, the composition of technical toxaphene as a function of the number of chlorine substituents was provisionally calculated and was found that hepta- and octachlorinated compounds represents 75% of the total toxaphene area. PMID- 12779229 TI - Mathematical representation of electrophoretic mobility of basic drugs in ternary solvent buffers in capillary zone electrophoresis. AB - The electrophoretic mobilities of two beta-blocker drugs, i.e., labetalol and atenolol, have been determined in a mixed solvent background electrolyte system containing sodium acetate+acetic acid as buffering agent and different volume fractions of water, methanol and ethanol using capillary electrophoresis. The produced data and three other sets collected from a recent work are employed to study the accuracy and prediction capability of a mathematical model to calculate the electrophoretic mobility with respect to the volume fractions of the solvents in the mixture. The results show that the proposed model is able to correlate/predict the mobility within an acceptable error range and it is possible to use the model in industry to achieve the optimum solvent composition for the buffer where using a ternary solvent system is required. The average percentage deviations (APDs) obtained for correlated and predicted data points are 0.71-2.48 and 1.72-4.39%, respectively. The accuracy of the proposed model is compared with that of a mixture response surface method and the results show that the proposed model is superior from both correlation and prediction points of view. The possibility of calculation of the mobility of chemically related drugs in water-methanol-ethanol mixtures using the proposed model is also shown and the produced prediction APD is approximately 8%. PMID- 12779230 TI - Determination of the absolute mobility and the equivalent ionic conductivity of NpO2+ at 25 degrees C and at infinite dilution by capillary electrophoresis inductively coupled plasma-mass spectrometry. AB - The absolute mobility of NpO2+ and its equivalent ionic conductivity were extrapolated at 25 degrees C and at infinite dilution using a set of experimental data obtained at various ionic strengths. The separation was carried out by capillary electrophoresis (CE) at various concentrations of creatinine at a pH of 5. The detection of NpO2+ was performed by inductively coupled plasma mass spectrometry coupled on-line with CE. The following values have been found: mu0NpO2+ (25 degrees C) = (2.94 +/- 0.07) x 10(-4) cm2 V(-1) s(-1) and lambda0NpO2+ (x 10(4), 25 degrees C) = 28.3 +/- 0.7 m2 S mol(-1). PMID- 12779231 TI - Determination of spore concentration in Bacillus thuringiensis through the analysis of dipicolinate by capillary zone electrophoresis. AB - A new capillary zone electrophoresis (CZE) method for the analysis of dipicolinic acid, a specific component found in spores but not in vegetative cells, was used to determine spore concentration in Bacillus thuringiensis according to the relationship between the spore concentration and the content of dipicolinate. The quantitative relationship was established by using purified spores. Electrolyte conditions that affected the separation efficiency of dipicolinate and the reproducibility were investigated. With 10 mM phosphate, 10 mM ethylenediaminetetraacetic acid and 0.25 mM tetradecyltrimethylammonium bromide at pH 6.2 as the carrier electrolyte, dipicolinate can be determined within 8 min at an applied voltage of -25 kV (anode at detector) and a capillary temperature of 25 degrees C. The method has a high separation efficiency with which the number of theoretical plates is above 300,000 plates m(-1). The relative standard deviations for migration time and peak area are less than 0.5% and 2.0%, respectively. The detection limit for dipicolinate was 10 ng ml(-1), which corresponds to 7.2 x 10(5) spores ml(-1). The method was used to determine spores in fermentation broths, and the results obtained agreed well with the values obtained by plate counting. PMID- 12779232 TI - Determination of biotin on a protein by quantitative sodium dodecyl sulfate capillary gel electrophoresis of monomeric avidin. AB - Sodium dodecyl sulfate-capillary gel electrophoresis (SDS-CGE) is performed to quantify monomeric avidin and biotin on a protein. Under non-reducing SDS-CGE conditions, avidin migrates as monomers exhibiting apparent molecular mass 17,000. In the presence of a biotin-protein conjugate, monomeric avidin binds the conjugate and forms a larger complex that migrates later in the separation. The difference between the remaining monomeric avidin and the initial amount is the portion of monomeric avidin bound to the conjugate. Accordingly, the number of biotin on the protein can be calculated. The assay is linearly responsive to increasing biotin loading in a biotinylation reaction of a protein. Accuracy of the assay is also demonstrated by good sample dilution recovery. Excellent quantitative reproducibility < 2% (relative standard deviation) is obtained for both intra- and inter-day measurements. Main advantages of the method include the use of monomeric avidin that minimizes steric hindrance to capture biotin on a protein and assay automation on a capillary electrophoresis apparatus. PMID- 12779233 TI - Determination of uracil in 5-fluorouracil substance by high-performance liquid chromatography. AB - Separation of 5-fluorouracil and uracil in chromatographic systems consisting of a silica column as a stationary phase and ethyl acetate or ethyl acetate partially saturated with water as a mobile phase has been studied. The results indicated that when saturation of ethyl acetate with water was chosen to reach more than 60%, such mobile phases [e.g., 2.4% (v/v) water in ethyl acetate] are useful in determining uracil in 5-fluorouracil substances. PMID- 12779234 TI - Enantiomeric separation of acidic compounds of pharmaceutical interest by capillary electrochromatography employing glycopeptide antibiotic stationary phases. AB - Enantiomeric separation of some selected acidic compounds of pharmaceutical interest belonging to the group of non-steroidal anti-inflammatory drugs were separated by capillary electrochromatography employing silica based glycopeptide antibiotic stationary phases, namely vancomycin or a teicoplanin derivatives (Hepta-Tyr). The vancomycin stationary phase allowed to achieve the chiral resolution of some racemic studied compounds only using mobile phases containing ammonium formate at a relatively low pH 2.5-3.5 and acetonitrile. Employing the teicoplanin derivative stationary phase, good enantiomeric resolution was achieved eluting with mobile phases containing sodium phosphate pH 6 acetonitrile. Enantiomers were moved to the detector because a relatively high reversed electroosmotic flow (due to the positive charge of the stationary phase) and to the electrophoretic mobility of analytes. PMID- 12779235 TI - Bacillus subtilis CPx-type ATPases: characterization of Cd, Zn, Co and Cu efflux systems. AB - Metal ion homeostasis requires the balanced expression of metal ion uptake systems, when metals are limiting, and corresponding efflux or storage systems, when metals are in excess. CPx-type ATPases are a family of membrane-bound transporters that often function to export toxic metals from cells. The Bacillus subtilis genome encodes three CPx-type ATPases: zosA, yvgW and yvgX. We demonstrate that yvgW and yvgX encode CadA and CopA, respectively, and that these genes function in metal ion resistance. A cadA mutant was sensitive to Cd(II), Zn(II) and Co(II), but not copper. Transcription of cadA initiates from a single, sigmaA-type promoter and was induced by Cd(II), Zn(II), and Co(II). The adjacent copZA operon is expressed as a bicistronic transcript from a sigmaA-type promoter and is selectively induced by copper. Mutation of either copZ, encoding a metallochaperone, or copA sensitizes the cells to copper but not to other metal ions. PMID- 12779236 TI - Identification and mutational studies of conserved amino acids in the outer membrane receptor protein, FepA, which affect transport but not binding of ferric enterobactin in Escherichia coli. AB - Many gram-negative bacteria produce and excrete siderophores, which complex iron with high affinity in the environment. The ferric siderophore complexes are transported across the outer membrane by receptor proteins. This process requires energy and is TonB dependent and must involve conformational changes in the receptor proteins to allow the transport of the ferric siderophores from the extracellular binding site to the periplasm. There is a large variety in the structures, molecular weights and charges among the siderophores. It was therefore realized that when the sequences of the many different receptor proteins were compared, simultaneously, all identities and close similarities, found in this manner, could only be due to residues involved in the conformational changes and transport mechanism, common to all the proteins, and not be due to the specificity of ligand recognition. Once the crystal structures of FepA, FhuA and FecA became available, it was immediately clear that the sequence similarities which were found in the simultaneous alignment, were all localized in a few structural domains, which are identical in the three structures and can therefore be expected to be maintained in all the proteins in this family. One of these domains, tentatively named the lock region, consists of 10 residues with a central quadrupole formed by two arginines and two glutamates, from the plug region and the beta barrel. We mutated several of these residues in FepA. All showed normal binding in quantitative binding studies. Some showed normal transport as well, however, the majority showed moderate to severe defective transport with ferric enterobactin. The results therefore show the validity of the hypothesis that the simultaneous sequence alignment will select the residues involved in the transport function of the receptor proteins. In addition the results allow to relate the severity of the transport deficiency to be correlated with the structure of the lock region while it is also possible to propose a function of this region in the conformational changes of the protein during the transport of the ligand from the binding site to the periplasm. PMID- 12779237 TI - Variability of the Fenton reaction characteristics of the EDTA, DTPA, and citrate complexes of iron. AB - The common metal chelation agents, DTPA and EDTA are often used as models for physiological low-molecular weight iron complexes in biochemical studies, or for common biochemical protocols. In the biochemical literature there are apparent conflicts as to whether EDTA and DTPA are pro-oxidant or antioxidant additives. This apparent conflict is puzzling since in chemical systems FeIIEDTA and FeIIDTPA are well known Fenton reaction reagents. In this investigation we examined the voltammetric characteristics of the iron complexes of EDTA, DTPA, and citrate and the effect of the ligand:metal ratio (L:M) on the electrocatalytic (EC') waves that result from reduction of H2O2 by this complex. At a ratio of 1:1, the cyclic voltammetric waves of the complexes indicate the presence of a reversible species corresponding to the Fe(II/III)L couple, along with a second irreversible reduction peak. The second irreversible voltammetric peak decreases at higher L:M ratios for EDTA and citrate. The 1:1 iron complexes of EDTA, DTPA, and citrate clearly induce the catalytic reduction of H2O2. In the presence of a greater than 100 fold excess of H2O2 relative to iron, higher L:M ratios greatly reduced the catalytic EC' wave compared to the 1:1 ratios. At H2O2:Fe ratios less than 50, the L:M ratio has very little effect of the EC' current. These observations may explain the apparent discrepancies in the biochemical literature. Addition of EDTA or DTPA may enhance oxidative processes if the L:M is low (less than unity), whereas rates of on-going oxidative processes may decrease if that ratio, along with the relative amount of H2O2, are both high (excess ligand). The impact of this study is of particular importance given the widespread use of these ligands in biochemical studies. PMID- 12779238 TI - Cobalt-resistance in wall-less mutant (fz; sg; os-1) of Neurospora crassa. AB - A cobalt-resistant wall-less mutant (slime) of Neurospora crassa was obtained by repeated sub-culturing of the sensitive wall-less mutant (W-sl) on agar medium containing toxic concentrations of cobalt. Resistance was stable on culturing Cor sl on cobalt-free medium up to 15 weekly subcultures. Cor-sl is 10-fold more resistant to cobalt when compared to W-sl. It is also cross-resistant to Cu (10 fold) and Ni (3-fold). Cobalt accumulated by Cor-sl during growth and in short term uptake experiments was lower when compared to W-sl. Cells previously loaded with cobalt was released into medium in both mutants, while in case of Cor-sl most of cobalt taken up (> 80%), was released back into the medium when compared to W-sl. Metabolic inhibitor (Sodium azide) and magnesium ions inhibited cobalt uptake in both the mutants. Fractionation of cell-free extracts showed that most of the cobalt (70%) taken up by Cor-sl was bound to an inducible protein fraction which bound to DEAE-Cellulose, while in W-sl only 20% of cobalt was associated with this fraction. Subcellular localization of cobalt in W-sl indicated most of it to be cytoplasmic (70%) while nuclei and mitochondria had 10% and 5% respectively. In case of Cor-sl, mitochondrial cobalt accounted for only 2% while no significant differences were noted for other fractions. Our data implicate both transport block and intracellular sequestration of cobalt to play a major role in resistance. PMID- 12779239 TI - Trishydroxamates and triscatecholates based on monosaccharides and myo-inositol as artificial siderophores. AB - New trishydroxamates and triscatecholates based on methyl alpha-D glucopyranoside, methyl alpha-D-galactopyranoside, methyl alpha-D-ribopyranoside and methyl alpha-D-xylopyranoside as well as on 1,3,5-tri-O-benzyl-myo-inositol were synthesized. N-Methylsuccinohydroxamate, N-methylglutarohydroxamate and their O-benzoyl derivatives were used as hydroxamate moieties. 2,3 Dihydroxybenzoyl derivatives and acylated compounds as well as 2,3- and 3,4 dihydroxybenzylidenehydrazino derivatives, partly with spacer groups, were utilized as catecholate components. The siderophore activity of the prepared siderophore analogues was examined by a growth promotion assay with various Gram negative bacteria and mycobacteria and by the CAS-assay. Some trishydroxamates and triscatecholates showed siderophore activity on Gram-negative bacteria and triscatecholates on mycobacteria. Iron complexes of the trishydroxamates act as siderophores for all types of iron transport mutants. The recognition and uptake specificity of these compounds was studied by E. coli siderophore receptor and iron transport mutants. Structure activity correlations are discussed. PMID- 12779240 TI - Cis-[Pt(Cl)2(pyridine)(5-SO3H-isoquinoline)] complex, a selective inhibitor of telomerase enzyme. AB - Since it has been widely demonstrated that platinum-based drugs, like cisplatin, carboplatin and oxaliplatin, bind preferentially to guanine in N7 position and that telomerase assemblage includes a RNA portion rich in guanine, we previously designed and synthesized a series of new complexes with a cytotoxic [Pt(II)Cl2] moiety, with the aim of selecting carrier ligands able to inhibit telomerase enzyme. Among these compounds, [cis-dichloropyridine-5-isoquinolinesulfonic acid Pt(II)], named Ptquin8, showed the most significant inhibition of telomerase in a cell-free biochemical assay. In this paper, we report the biological effects of Ptquin8 on in vitro tumor model (MCF-7). This complex is able to reduce telomerase activity from 12 to 46%, in a concentration range between 10(-9) and 10(-5) M after 24 h continuous treatment. Moreover, Ptquin8 shows significant cytotoxicity after 10 days of continuous treatment only at concentrations higher than 10(-5) M. The determination of residual telomere length confirmed the inhibition of telomerase action. This induced a progressive reduction of the cell proliferative capacity, and the appearance of an elevated number of apoptotic cells after 18 days. RT-PCR analysis of telomerase RNA components excluded any interaction of the compound at genomic level. The biochemical effects of Ptquin8 were also evaluated on non-neoplastic NIH3T3 cells, that are able to down regulate telomerase activity as a consequence of the confluence contact inhibition. In this cell model, the reactivation of telomerase due to re-seeding at lower density was significantly inhibited by Ptquin8 in a dose-dependent manner. These results highlight a possible role of Ptquin8 as a selective anti telomerase tool for cancer treatment. PMID- 12779241 TI - Long lasting cadmium intake is associated with reduction of insulin receptors in rat adipocytes. AB - The effects of chronic cadmium exposure on adipose tissue have not been extensively reported. In adult Wistar male rats we investigated in vivo effect of 6 weeks lasting cadmium intake in drinking tap water (CdCl2 9,7 mg/l). Insulin receptors in isolated adipocytes from epididymal fat and glucose transporter protein GLUT4 content in fat tissue plasma membranes were determined. Control and Cd treated rats had similar water intake with subsequent heavy augmentation of Cd content in liver of experimental animals. In comparison with controls, Cd intake did not influence body mass increment and fat cell size, but significantly increased serum glycemia and moderately elevated insulinemia. Cadmium intake significantly reduced (approximately 50%) both, total insulin receptors number and density of the receptors in fat cells. No differences in the content of GLUT4 in crude plasma membranes of adipose tissue were observed. Diminished insulin receptors in adipocytes could account for diabetogenic effect of long lasting cadmium intake. PMID- 12779242 TI - Nickel removal from nickel plating waste water using a biologically active moving bed sand filter. AB - Efficient removal of dissolved nickel was observed in a biologically active moving-bed 'MERESAFIN' sand filter treating rinsing water from an electroless nickel plating plant. Although nickel is fully soluble in this waste water, its passage through the sand filter promoted rapid removal of approximately 1 mg Ni/l. The speciation of Ni in the waste water was modelled; the most probable precipitates forming under the conditions in the filter were predicted using PHREEQC. Analyses of the Ni-containing biosludge using chemical, electron microscopical and X-ray spectroscopic techniques confirmed crystallisation of nickel phosphate as arupite (Ni3(PO4)2 x 8H2O), together with hydroxyapatite within the bacterial biofilm on the filter sand grains. Biosorption contributed less than 1% of the overall sequestered nickel. Metabolising bacteria are essential for the process; the definitive role of specific components of the mixed population is undefined but the increase in pH promoted by metabolic activity of some microbial components is likely to promote nickel desolubilisation by others. PMID- 12779243 TI - High-resolution identification of mercury in particles in mouse kidney after acute lethal exposure. AB - Contamination of the food chain by mercury is a major concern of Public Health of our day. Kidney and nervous system are the major targets of mercury toxicity in mammals. We show here that the detailed subcellular in vivo topography of microparticles of mercury in tissues can be achieved by scanning electron microscopy (SEM) coupled with X-ray elemental microanalysis (XRM). SEM-XRM offered the fine topography of mercury in the kidney of BALB/c mice that were submitted to an intraperitoneal lethal injection of mercuric chloride (HgCl2). All of the renal mercury was seen inside blood vessels located in both cortex and medulla of the mouse kidney. This blood-born mercury was organised in spheroid particles of less than 50 nm in diameter (31.4 +/- 14.1 nm). They were seen attached either to aggregates of plasma proteins or to the surface of blood cells. No evidence of internalisation of mercury by blood, endothelial or kidney cells was found. The average kidney density of mercury microspheres was 1920 +/- 1320 particles per mm2. We propose SEM-XRM as an elective approach to further investigations, at the subcellular level, on the quantitative dynamics of mercury particles in the tissues. PMID- 12779244 TI - Production of singlet oxygen on irradiation of a photodynamic therapy agent, zinc coproporphyrin III, with low host toxicity. AB - Zinc-coproporphyrin III (Zincphyrin) acts efficiently as a photodynamic therapy (PDT) agent in mice, while it shows no tumor cell-killing activity in vitro and has a high LD50 (low toxicity) in mice. It appears to have advantages over other porphyrins as a practical PDT reagent. In order to examine the action mechanism of Zincphyrin in PDT, we evaluated the photochemical characteristics of Zincphyrin by measurement of the near-infrared emission at 1268 nm, which provides direct evidence for formation of 1O2. Intense emission was observed in the presence of Zincphyrin, and was completely inhibited by NaN3, a 1O2 scavenger. Based on a quenching study, the rate constant of the reaction of 1O2 with NaN3 was determined to be 1.5-3.5 M(-1) s(-1), which is close to the reported value (3.8 x 10(8) M(-1) s(-1)). The intensity of the 1O2-specific emission was proportional to both the laser power and the concentration of Zincphyrin. The fluorescence quantum yield of Zincphyrin was 0.004 in phosphate buffer (100 mM, pH 7.4), which indicates that the excited state decays via other pathway(s) faster than through the fluorescence emission pathway. The lifetime of the triplet state of Zincphyrin (210 micros) was relatively long compared to that of other porphyrins, such as hematoporphyrin (Hp) (40 micros), coproporphyrin I (50 gs), or coproporphyrin III (36 gs). These results demonstrate the photodynamic generation of 1O2 by Zincphyrin. PMID- 12779245 TI - The HET-CAM test combined with histological studies for better evaluation of active ingredient innocuity. AB - In the search for alternative methods to animal testing, the Hen's egg test on chorioallantoic membrane (HET-CAM) plays a central role in evaluating the innocuity of active ingredients. Therefore, in the following studies we combined the HET-CAM test with histological evaluation in order to increase the sensitivity of evaluation. Twenty active ingredients from four different categories of origin (vegetal, marine, biotechnological and chemical synthetic) were subjected to innocuity evaluation at two different concentrations (pure and 10%). We performed the HET-CAM test and histological evaluation after trypan blue and hematoxylin-eosin staining of the chorioallantoic membrane to microscopically evaluate its state of damage after application of each active ingredient. These studies showed that when the active ingredient was diluted (10%), no discrepancy was seen between the classical HET-CAM evaluation and the histological reading of the chorioallantoic membrane. The histological findings corresponded with the visual observation of the CAM. When the active ingredients were tested at pure concentration, 7 out of 20 tested products demonstrated discrepancy between the two tests. In six cases, the histological examination revealed signs of irritation, such as hyperemia, while visual HET-CAM evaluation was negative. In another case, the histological examination revealed a slight hemorrhage whereas the HET-CAM reading showed only hyperemia. Moreover, the results of trypan blue staining corroborated the histological evaluation of the CAM. These results strongly suggest that the combination of histological and visual HET-CAM tests is of interest for a more sensitive evaluation of the innocuity of cosmetic active ingredients. This additional sensitivity may help to prevent some cases of in vivo intolerance reactions. PMID- 12779246 TI - Topical retinaldehyde treatment in oral lichen planus and leukoplakia. AB - The aim of this exploratory study was to assess the efficacy of a natural metabolite of vitamin A, retinaldehyde 0.1%, vehicled in a gel in 17 patients with oral lichen planus and in 13 patients with oral leukoplakia, twice daily for 2 months. Our investigation was clinical, histological, immunohistochemical through the expression of markers of cell terminal differentiation and biochemical by using two-dimensional gel electrophoresis of cytokeratins (CK). In addition, the activity of retinaldehyde was studied ex vivo on surviving buccal mucosa. Retinaldehyde gel 0.1% showed good clinical efficacy, resulting in 6% disappearance and 82% improvement of the lesions in lichen planus and 17% disappearance and 75% improvement in leukoplakia. This was confirmed with immunohistochemistry, which revealed down-regulation of filaggrin and CK-10 as markers of terminal differentiation in both diseases. The effects of retinaldehyde in these two diseases were further demonstrated in the ex vivo surviving mucosal model, resulting in histological disappearance of keratinization in 80% of the lichen planus fragments and 40% of the leukoplakia fragments, associated with down-regulation of filaggrin and CK-10. PMID- 12779247 TI - Date palm kernel extract exhibits antiaging properties and significantly reduces skin wrinkles. AB - Hormones play a central role in skin appearance and are implicated in skin aging. Recently, along with the remarkable increase in interest in natural products, the application of phytohormones in antiaging products has become very important. In this context, we developed date palm kernel extract. Date palm kernel is rich in phytohormones and we investigated the antiaging properties of date palm kernel in this in vivo study on wrinkles. Ten healthy women volunteers, between the ages of 46 and 58 years, applied the cream formula with 5% date palm kernel or placebo on the eye area twice a day for 5 weeks. The evaluation was made both clinically and by silicon replica analysis followed by statistical analysis using the Wilcoxon test. Silicon replica results showed that topical application of date palm kernel reduced the total surface of wrinkles by 27.6% (p = 0.038). Moreover, date palm kernel reduced the depth of wrinkles by 3.52% (p = 0.0231). These results are statistically significant and were clinically confirmed where visual improvement was seen in 60% of the volunteers treated. This in vivo study demonstrates that date palm kernel exhibits a significant antiwrinkle effect and is therefore of interest in antiaging skin care products. PMID- 12779249 TI - Inhibitory effect of local ischemic preconditioning on gamma ray-induced lipid peroxidation in rats: a preliminary study. AB - We examined the effect of local ischemic preconditioning on postradiation lipid peroxidation in the serum of total body irradiated rats. Markers of peroxidative damage provoked by radiation alone or radiation preceeded by ischemic preconditioning were thiobarbituric acid reactive substances, triglycerides and uric acid concentrations in serum. These data indicated that local ischemic preconditioning modifies the peroxidizing effects of radiation through inhibition of free radical-dependent lipid peroxidation. Other unrecognized mechanisms are probably also involved. Uric acid could act as an antioxidant against radiation alone and local preconditioned ischemia together with radiation. PMID- 12779248 TI - Up-regulation of vascular endothelial growth factor and down-regulation of pigment epithelium-derived factor messenger ribonucleic acid levels in leptin exposed cultured retinal pericytes. AB - Leptin, a circulating hormone secreted mainly from adipose tissues, is involved in the control of body weight. Recently, leptin was found to be an angiogenic factor and its vitreous levels were shown to be elevated in patients with angiogenic eye diseases such as proliferative diabetic retinopathy. However, the role of leptin in diabetic retinopathy is not fully understood. Since pericyte loss and dysfunction have been considered to be one of the characteristic changes of the early phases of diabetic retinopathy, we investigated the effects of leptin on the growth and function of bovine cultured retinal pericytes. Although it did not affect cell growth, leptin significantly up-regulated pericyte messenger ribonucleic acid levels of an endogenous angiogenic stimulator, vascular endothelial growth factor (VEGF). Leptin was also found to significantly inhibit gene expression of pigment epithelium-derived factor (PEDF), the most potent angiogenesis inhibitor in the mammalian eye, in pericytes. The present study suggests that leptin might elicit angiogenesis through VEGF induction as well as PEDF suppression in pericytes and could thus be involved in the development and progression of diabetic retinopathy, especially in obese insulin resistant patients. PMID- 12779250 TI - Drosophila neuropeptide signaling. PMID- 12779251 TI - From genes to aging in Drosophila. AB - Despite the intimate nature of the aging process we actually know little about it. In more recent years, work on a variety of organisms, utilizing approaches including demography, molecular genetics, and epidemiology, have challenged some of the more commonly held assumptions about the aging process. These studies have served to reinvigorate the field of aging research and are beginning to lead the way in a renaissance in aging research (Helfand and Inouye, 2002). Invertebrate model systems such as Drosophila and Caenorhabditis elegans that permit extensive genetic analysis are at the forefront of this renaissance. PMID- 12779252 TI - Genetics of wheat gluten proteins. PMID- 12779253 TI - Pathology quiz 43. PMID- 12779254 TI - Quality of life following surgical treatment of oculopharyngeal syndrome. AB - Oculopharyngeal syndrome is a hereditary disease mostly affecting French Canadians. Cricopharyngeal myotomy and blepharoplasty are commonly performed on these patients. We reviewed the files of 20 patients who underwent these procedures between June 1986 and June 1999. The average perioperative stay was 4.7 days. The nine patients still alive were contacted and answered a telephone survey on their quality of life. None reported postoperative complications. Myotomy significantly improved the medical domain of their quality of life as confirmed by improvement in the dysphagia for solids, food incarceration, duration of meals, and choking during meals. The psychological domain also improved, namely, appetite and interest in food. The effects of blepharoplasty were also notable. Long-term satisfaction is reported by the majority of patients up to a maximum of 10 years postoperatively. In conclusion, simple procedures such as blepharoplasty and cricopharyngeal myotomy greatly improved the quality of life of these patients. We therefore recommend early interventions. PMID- 12779255 TI - Endoscopic septoplasty: technique and outcomes. AB - Recent literature has already embraced the topic of endoscopic septoplasty, with several published articles on the subject. This approach provides a direct targeted route to the anatomic deformity, improved visualization, and magnification of the surgical field. It allows improved evaluation of the posterior nasal septal deformities, identification of the degree of mucosal involvement of the posterior ends of the inferior turbinates, and concomitant assessment of the middle meatus. It permits objective documentation of the cause of nasal obstruction with possible use in outcome assessment. It is also an effective teaching method and a motivating approach for the nursing team. We present our experience in a series of 47 patients performed during a 1 1/2-year period and discuss the surgical technique and patients' outcomes. We systematically used the endoscope for all septal and turbinate surgery. We evaluated outcomes using a telephone survey along with a validated disease specific health status measure and a global rating questionnaire. PMID- 12779256 TI - Variability in the vascularity of the pectoralis major muscle. AB - OBJECTIVE: Although the pectoralis major muscle has been the subject of numerous anatomic studies over the past 20 years, there remains a high complication rate for pedicled pectoralis musculocutaneous flaps. In this report, angiograms of 43 pectoralis major muscles were studied to assess the vascular territories of its three arterial supplies: lateral thoracic artery, the pectoral branch of the thoracoacromial artery, and the anterior intercostal perforators of the internal mammary artery. METHODS: Twenty-two adult human cadavers underwent whole-body arterial perfusion (200 mL/kg) with a mixture of lead oxide, gelatin, and water through the carotid artery. All pectoralis major muscles were dissected and radiographed. Radiographs were photographically printed as contact prints. The vasculature of each muscle was analyzed using the paper template technique. RESULTS: The pectoral branch of the thoracoacromial artery supplied 50.7% of the vascular territory of the pectoralis major. The lateral thoracic artery was present in 37 of 43 angiograms and supplied a mean territory of 6.6%. The anterior intercostal perforating branches of the internal mammary artery supplied 43% of muscle parenchyma. There was considerable variability in the extent of various vascular territories from muscle specimen to specimen. CONCLUSION: Despite excellent surgical technique, certain pedicled musculocutaneous pectoralis major flaps may suffer partial distal necrosis simply owing to the relatively small vascular territory of the pectoral branch of the thoracoacromial artery. PMID- 12779257 TI - Survey of the use of suction drains in head and neck surgery and analysis of their biomechanical properties. AB - BACKGROUND: Closed suction drains have an important role in surgical wound healing. Although most surgeons use them routinely, indications for use and their postoperative management (emptying, removal) vary. The purpose of this study was to assess drain use by head and neck surgeons in Canada, to conduct a biomechanical analysis of the drains in a laboratory setting, and to make recommendations for drain use and management. METHODS: A survey was mailed to 343 active members of the Canadian Society of Otolaryngology. Three sets of experimental trials were conducted on the most commonly used drains to assess the effect of increased reservoir filling on suction generated through (1) incrementally increasing the amount of fluid within the reservoir, (2) compression of the reservoir with no fluid within, and (3) compression with the reservoirs while filled to 25% capacity with fluid. RESULTS: A 41% response rate was obtained. It was found that the majority of head and neck surgeons in Canada use Hemovac and Jackson-Pratt drainage systems routinely. There is considerable variability in practice with regard to drain emptying and timing of removal. Experimental results indicate that as filling of the reservoir increases, suction generated decreases sharply, to between 13 and 20% of initial values at 50% capacity. CONCLUSION: Postoperative drain management has important implications in surgical wound healing. Drain reservoirs should be monitored frequently to ensure adequate compression, particularly in the first 24 hours after insertion. Anticipated volume of drainage should dictate in part which reservoir is chosen. A larger reservoir is preferable in most cases. Drains should be removed promptly to decrease the risk of wound contamination. PMID- 12779258 TI - Hearing loss in acoustic neuromas following stereotactic radiotherapy. AB - BACKGROUND: Hearing preservation is a major concern in the management of acoustic neuroma (AN), whether treated by surgery or radiation or followed conservatively. OBJECTIVE: To assess the change in hearing in patients who underwent radiotherapy for ANs and to compare it with data from conservatively managed patients. DESIGN: Prospective case series. SETTING: Neuro-otology, neurosurgery, and radiation oncology practices in tertiary care hospitals of the University Health Network, University of Toronto. PATIENTS AND METHODS: Between 1991 and 1999, 32 patients who underwent radiotherapy were followed prospectively. Seventeen patients had pretreatment measurable hearing and a minimum 1-year follow-up. Twelve of these patients underwent Gamma Knife radiation and five had fractionated radiation therapy with a linear accelerator. Pre- and post-treatment hearing was classified according to the Gardner-Robertson (GR) classificatiom and according to the 1995 guidelines of the American Academy of Otolaryngology-Head and Neck Surgery Committee on Hearing and Equilibrium (AAO-HNS CHE). OUTCOME MEASURE: Change in hearing post-treatment. RESULTS: Mean follow-up was 48 months. Deterioration in the level of hearing was observed in 8 and 11 of the patients using the AAO-HNS CHE and the GR classifications, respectively. Forty-four to 70% of the patients had lost their pretreatment serviceable hearing, depending on the classification of hearing used and how serviceable hearing was defined. Compared with series of patients followed conservatively in our institution and in other studies, patients who receive stereotactic radiation appear to lose hearing at least at the same rate. CONCLUSION: Our data suggest that hearing loss continues to occur despite radiotherapy, and patients should be advised accordingly. PMID- 12779259 TI - Langerhans' cell histiocytosis in the paediatric population: presentation and treatment of head and neck manifestations. AB - Langerhans' cell histiocytosis (LCH) is a rare paediatric disease of unknown etiology affecting 1 to 5 children per 1 million each year. It is characterized by the idiopathic proliferation of Langerhans' cells. The clinical spectrum of disease is quite varied, ranging from a solitary eosinophilic granuloma to diffuse multisystem involvement. The head and neck is the most common site of involvement, occurring in approximately 60% of LCH patients. Head and neck manifestations are diverse and include skull and temporal bone lesions, cervical lymphadenopathy, and skin rash. Diagnosis can be difficult as these lesions mimic other common conditions seen by the otolaryngologist, including otitis externa, acute mastoiditis, and gingivitis. A retrospective study was carried out to study our centre's experience with LCH over the last 10 years. Twenty-one patients were diagnosed between January 1990 and December 1999. Patient's age at time of diagnosis ranged from 6 days to 14 years. Fifty-seven percent of patients had localized bony lesions; the remaining 43% had diffuse multisystem disease. The head and neck was also the most commonly involved site in our study, affecting 67% of our patients. Presentation and diagnosis of these lesions are discussed in detail. Treatment, complications, and patient outcomes will also be discussed. PMID- 12779260 TI - [Revision stapedectomy for otosclerosis: report of 73 cases]. AB - OBJECTIVE: The aim of this study was to determine causes and evaluate results of revision stapedectomy. DESIGN: Retrospective review of 73 revision stapedectomies. SETTING: Revision stapedectomies were performed in two tertiary otolaryngology departments (Fondation Adolphe de Rothschild, Paris, France, and Cliniques universitaires Saint-Luc, Bruxelles, Belgium). METHODS: Patients characteristics, indications for revision, intraoperative findings, and hearing results were noted. MAIN OUTCOME MEASURES: Hearing results were reported as recommended by the American Academy of Otolaryngology-Head and Neck Surgery. We also reported hearing results with and without the use of a laser. RESULTS: Conductive hearing loss was the main indication for revision (78%). Mean intraoperative findings included prosthesis malfunction (50%), fibrous adhesions (32.8%), incus necrosis (8.5%), and otosclerotic regrowth (7%). When revision was performed because of cochleovestibular complication, middle ear exploration revealed three findings: oval window granuloma (30.7%), perilymphatic fistula (30.7%), and a too long prosthesis (23%). Postoperative air-bone gap (ABG) was closed to less than 10 dB in 51.5% of cases and to less than 20 dB in 68.7% of cases, with 9% of sensorineural hearing loss (not exceeding 15 dB in 80% of cases). The use of an argon laser in 14 patients (19%) showed slightly better hearing results (postoperative ABG < 10 dB in 61.5% of cases and < 20 dB in 77% of cases), but this difference was not statistically significant compared with patients operated on without the use of a laser. CONCLUSION: The results of this series are comparable with previously published studies. Revision stapes surgery is not as successful as primary stapedectomy, but good gap closure can be expected in two-thirds of cases with an experienced surgeon. PMID- 12779261 TI - Expression of p53 in inverted papilloma and malignancy associated with inverted papilloma. AB - OBJECTIVE: To determine the association of p53 protein in malignant cases of inverted papilloma compared with cases of benign inverted papilloma of the sinonasal tract. DESIGN: Case-control study of archived pathologic material. SETTING: Tertiary care hospital. METHODS: Archived pathologic material of cases of malignancy associated with inverted papilloma and controls of benign inverted papilloma were obtained from Mount Sinai Hospital. These were subjected to immunohistochemistry for p53. Clinical correlation was obtained by retrospective chart review. MAIN OUTCOME MEASURES: Staining of pathologic specimens for p53 and survival or recurrence. RESULTS: Four of the five cases of malignancy associated with inverted papilloma demonstrated overexpression of p53. None of the benign cases of inverted papilloma demonstrated overexpression. Only two of the five patients with malignancy associated with inverted papilloma were alive at 2 years. CONCLUSIONS: Overexpression of p53 may serve as a marker for malignant transformation of inverted papilloma. PMID- 12779262 TI - Surgical management of nonmalignant parotid masses in the pediatric population: the Montreal Children's Hospital's experience. AB - Nonmalignant parotid masses in children can have protean etiologies ranging from infective parotitis to a benign neoplastic, vascular, or congenital origin. We review the 10-year experience of a tertiary care pediatric centre with respect to the surgical management of nonmalignant parotid masses. In total, 15 patients with nonmalignant masses of the parotid gland region underwent surgery. Five children were diagnosed with lymphoepithelial cyst or first branchial cleft cyst. Three children were diagnosed with parotid abscess, one of whom had atypical mycobacteria. Other diagnoses included lymphangioma (three cases), chronic inflammation (two cases), and epidermoid cyst (one case). One patient who presented with a parotid cyst was diagnosed postoperatively with plexiform neurofibroma of the facial nerve. She was the only patient with postoperative facial nerve paresis, affecting the orbital branch. Presentation and postoperative complications of these surgically managed nonmalignant parotid masses are reviewed. The history and physical examination are of the utmost importance in predicting the diagnosis, although ultrasonography and computed tomography can be useful. Fine-needle aspiration cytology was not well tolerated by children and appears of little use as the accurate diagnosis was provided by the surgical pathology specimen. PMID- 12779263 TI - Early complications of tracheostomy performed in the operating room. AB - Tracheostomy is one of the oldest operations in medicine. The intraoperative and postoperative complications associated with this procedure are well established. Recently, percutaneous and open bedside tracheostomy in the intensive care unit has been reported as an alternative to tracheostomies performed in the operating room. We investigated the early complications in 70 consecutive tracheostomies performed in the operating room in Bnai Zion Medical Center in Haifa, Israel. The study revealed no complications related to patient transportation to and from the operating room. The complication rate of standard surgical tracheostomy performed in the operating room was very low. PMID- 12779264 TI - Jejunojejunal intussusception: an unusual cause of free radial forearm flap necrosis following pharyngolaryngectomy. PMID- 12779265 TI - Cervical cystic schwannoma of the vagus nerve: diagnostic and surgical challenge. PMID- 12779266 TI - Sebaceous gland carcinoma of the scalp: case report and review of the literature. PMID- 12779267 TI - Nasopharyngeal choristoma: case report and review of the literature. PMID- 12779268 TI - Potential myocardial iron content evaluation by magnetic resonance imaging in thalassemia major patients treated with Deferoxamine or Deferiprone during a randomized multicenter prospective clinical study. AB - The purpose of this study was to evaluate if the variations of heart magnetic resonance imaging in beta-thalassemia major patients treated with Deferoxamine B mesylate (DF) or Deferiprone (L1) chelation therapy is a useful tool of the indirect myocardial iron content determination. For this reason, a prospective study was carried out. Seventy-two consecutive patients with beta-thalassemia major (35 treated with DF and 37 with L1) were studied. The main outcome results were laboratory parameters including determination of the liver iron concentration (LIC) and magnetic resonance imaging (MRI) of the heart and liver. The heart to muscle signal intensity ratios (HSIRs) were significantly increased in both the DF (t = -2.8; p < 0.01) and L1 (t = -3.1; p < 0.01) groups after one year of treatment No statistically significant difference in the values of HSIRs was present between the two groups at the beginning of treatment (p = 0.25; t = 1.13), and after one year of treatment (p = 0.20; t = 1.28). The HSIR were inversely correlated to the LIC (r = -0.52; p < 0.001) but not with ferritin levels (r = 0.10; p = 0.18). A positive correlation was found between the variation of HSIRs and that of the liver signal intensity ratios (r=0.52; p < 0.001), and a mild correlation (r = 0.40; p < 0.001) was found between the gamma glutamyltransferase (gammaGt) levels and the HSIRs values. Our data confirm that heart MRI is sensitive enough to detect significant variations of the mean HSIR during iron chelation with DF or L1. PMID- 12779269 TI - Flow cytometric analysis of hydroxyurea effects on fetal hemoglobin production in cultures of beta-thalassemia erythroid precursors. AB - An increase in fetal hemoglobin (Hb F) ameliorates the clinical symptoms of the underlying disease in the beta hemoglobinopathies-sickle cell anemia and beta thalassemia (thal). Hydroxyurea (HU) can elevate Hb F production in erythroid cells and is the agent currently in clinical use for patients with sickle cell anemia; it is presently being tested in clinical trials for thalassemia. We have developed a two-phase liquid culture system that mimics the in vivo hematological changes that are observed in patients treated with HU. Adding HU during the second phase of the culture increases the proportion of Hb F, increases the levels of total hemoglobin (Hb) content per cell and increases cell size, but it decreases the numbers of cells and the total amount of Hb produced. In the present study we developed and utilized a double labeling procedure for flow cytometric analysis of the cellular distribution of Hb F. Cells exposed to various concentrations of HU on day 6 of the second phase of the culture were harvested on day 12, and stained simultaneously with fluorochrome-conjugated monoclonal antibodies specific for human glycophorin A, an erythroid specific marker, and human Hb F. Both the percentage of the Hb F-containing cells and their intensity of fluorescence were recorded. The latter value gives a semi quantitative estimation of the mean cellular Hb F content. The results indicated that cultures derived from different beta-thalassemic patients differ in their response to HU. In most patients, low doses of HU decreased the percentage of Hb F-cells as well as their Hb F content. At high doses, some patients showed an increase in both parameters, while others showed an increase in the percentage of Hb F-cells with minimal increase in their mean Hb F content, while still other patients showed little effect at all. In all patients, high doses of HU caused a decrease in cell numbers. These results suggest that HU has mixed effects on erythroid precursors. Both the two-phase liquid culture and the flow cytometric analysis procedures described herein provide the experimental tools for screening of Hb F-inducing drugs and for evaluating patients' cell response prior to treatment. PMID- 12779270 TI - Analysis of beta-thalassemia mutations in northern Thailand using an automated fluorescence DNA sequencing technique. AB - A total of 218 beta-thalassemia (thal) genes from 109 beta-thal major patients were characterized using an automated fluorescence DNA sequencing technique. Eight different mutations were identified in all 218 alleles (100%). Four common mutations accounted for 96.8% [49.5% were codons 41/42 (-TTCT), 34.4% were codon 17 (A --> T), 6.9% were IVS-I-1 (G --> T) and, 6.0% were codons 71/72 (+A)]. There were three cases of -28 (A --> G) and one of IVS-II-654 (C --> T), mutations that have been previously described in Thai subjects. We also identified two mutations in the beta-globin promoter region which have not been reported in Thailand before [-31 (A --> G) and -87 (C --> A)]. Although these mutations are described as beta+-thal, the compound heterozygote with one of the common beta(o)-thal mutations exhibits the phenotype of beta-thal major. The frequency of beta-thal genes in northern Thailand were similar to the northeastern region, but different from those reported in southern and central Thailand, where IVS-I-5 (G --> C) and IVS-II-654 (C --> T) were the second most common anomalies, respectively. The spectrum of beta-globin gene mutations from this study will be useful for planning a prenatal diagnosis program especially for this region of Thailand. PMID- 12779272 TI - Origin of Hb A2' (Hb B2) [delta16(A13)Gly --> Arg (GGC --> CGC)]. AB - On a field trip to the Dogon country (le Pays Dogon) in central Mali, we detected a high frequency of the Hb A2 abnormality, reaching higher numbers among blacksmiths (up to 12.4%) living in the same villages. In this report, by direct nucleotide sequencing and employing a polymerase chain reaction-restriction fragment length polymorphism approach, we show that the Hb A2 variant observed in the Dogon population is indeed Hb A2', also called Hb B2, and that in all of the cases the abnormal delta-globin gene is linked to a unique haplotype. The same haplotype was found linked to Hb A2' in the Herero population belonging to the South African Bantu-speaking Blacks from Namibia. Although the unique origin of this mutation in Africa is a possibility, a recurrent mutational event cannot be excluded because the linked beta cluster haplotype is one of the two major haplotypes found in all African populations. A study of populations from other regions of Africa is required to clarify this issue. PMID- 12779271 TI - Molecular characterization of hereditary persistence of fetal hemoglobin in the Karen people of Thailand. AB - Hereditary persistence of fetal hemoglobin (HPFH) is the condition whereby a continuously active gamma-globin gene expression leads to elevated fetal hemoglobin (Hb F) levels in adult life [Stamatoyannopoulos G, Grosveld F. Hemoglobin switching. In: Stamatoyannopoulos G, Majerus PW, Perlmutter RM, Varmus H, eds. The Molecular Basis of Blood Diseases. Philadelphia: W.B. Saunders, 2001:135-182; Wood WG. Hereditary persistence of fetal hemoglobin and delta(beta) thalassemia. In: Steinberg MH, Forget BG, Higgs DR, Nagel RL, eds. Disorders of Hemoglobin: Genetics, Pathophysiology, and Clinical Management. Cambridge: Cambridge University Press, 2001:356-388; and Weatherall DJ, Clegg JB. Hereditary persistence of fetal hemoglobin. In: Weatherall DJ, Clegg JB, eds. The Thalassaemia Syndromes. Oxford: Blackwell Scientific Publishers, 1981:450-507]. The condition is caused either by mutation of the beta- and gamma-globin genes, or the gamma-gene controlled region on other chromosomes. Several families with this condition have been reported from Vietnam, Cambodia and China, and the Southeast Asian mutation (or HPFH-6), a 27 kb deletion, was demonstrated. Here we report on a mother and her daughter of the Karen ethnic group with high levels of Hb F, living in the Suan Pueng District on the border of Thailand and Myanmar. Genotyping showed a heterozygosity for the 27 kb deletion of the beta-globin gene. Their conditions have been confirmed by gap polymerase chain reaction (PCR) with three oligonucleotide primers recently developed by Xu et al. [Xu X-M, Li Z Q, Liu Z-Y, Zhong X-L, Zhao Y-Z, Mo Q-H. Molecular characterization and PCR detection of a deletional HPFH: application to rapid prenatal diagnosis for compound heterozygotes of this defect with beta-thalassemia in a Chinese family. Am J Hematol 2000; 65:183-188.], and a DNA sequencing method. Thus far there has been no official report of the HPFH-6 anomaly from Thailand. The compound heterozygosity of beta-thalassemia (thal) and hereditary persistence of Hb F causes the phenotype of thalassemia intermedia; in contrast, homozygotes for this anomaly show only mild microcytic anemia. Hence, genetic counseling for hereditary persistence of Hb F carriers is needed for family planning. PMID- 12779273 TI - A new unstable alpha2-globin gene variant: Hb Chartres [alpha33(B14)Phe --> Ser]. PMID- 12779274 TI - A new hemoglobin variant, Hb Fukui [alpha139(HC1)Lys --> Asn (AAA --> AAC) (alpha2)]. PMID- 12779275 TI - The mutation of Hb Turriff [alpha99(G6)Lys --> Glu (AAG --> GAG)] is carried by the alpha1-globin gene in a Japanese (Hb Turriff-I). PMID- 12779276 TI - Identification of alpha-thalassemia mutations in Iranian individuals with abnormal hematological indices and normal Hb A2. PMID- 12779277 TI - Co-existence of the codon 16 (-C) (beta(o)) and codon 10 (C --> A) (beta+) mutations on the same beta-globin gene. PMID- 12779278 TI - Hb St. Luke's [alpha95(G2)Pro --> Arg (alpha1)]. PMID- 12779279 TI - Design and evaluation of matrix diffusion controlled transdermal patches of verapamil hydrochloride. AB - Transdermal patches of verapamil hydrochloride were prepared using four different polymers (individual and combination): Eudragit RL100 (ERL100), Eudragit RS100 (ERS100), hydroxypropyl methylcellulose 15 cps (HPMC), and ethyl cellulose (EC), of varying degrees of hydrophilicity and hydrophobicity. The effect of the polymers on the technological properties, i.e., drug release, water vapor transmission rate (WVTR), and percentage moisture loss (ML), percentage moisture absorption (MA), folding endurance, and thickness, was investigated. Different formulations were prepared in accordance with the 2(3) factorial design, with ERL100 being the parent polymer. The patch containing ERL100 alone showed maximum WVTR, % MA, and % ML, which could be attributed to its hydrophilic nature. As expected, substitution with ERS100, HPMC, and EC decreased all the above values in accordance with their decreasing degree of hydrophilicity. In vitro release studies showed zero-order release of the drug from all the patches, and the mechanism of release was diffusion mediated. Moreover, the release of the drug was sustained and it extended over a period of 24 hr in all formulations. A12 emerged as the most satisfactory formulation insofar as its technological properties were concerned. Further, release and permeation of the drug from the most satisfactory formulation (A12) was evaluated through different biological barriers (shed snake skin, rabbit skin, and rat skin) to get an idea of the drug permeation through human skin. Shed snake's skin was found to be most permeable (82.56% drug release at 24 hr) and rat skin was least permeable (52.38%). Percutaneous absorption studies were carried out in rabbits. The pharmacokinetic parameters calculated from blood levels of the drug revealed a profile typical of a sustained release formulation, with the ability to maintain adequate plasma levels for 24 hr. [AUC: 3.09 mg/mL hr, Cmax: 203.95 microg/mL, Tmax: 8 hr]. It can therefore be concluded that the patch containing ERL100 and HPMC in the ratio 8:2 has achieved the objectives of transdermal drug delivery system, such as avoidance of first pass effect, extended release, and reduced frequency of administration. PMID- 12779280 TI - Formulation and efficacy studies of new topical anesthetic creams. AB - Local anesthetics (lidocaine or tetracaine) spontaneously melted at 25 degrees C when mixed with thymol and aqueous isopropyl alcohol solution (IPA) at proper ratios and formed novel two-phase melt systems (TMS). The TMS consisted of a homogeneous oil phase containing primarily a local anesthetic agent (lidocaine or tetracaine) and thymol, and a homogeneous aqueous phase containing primarily IPA and pH 9.2 buffer. The relationship between melting of the solid components and system composition was determined from the phase diagram obtained by a titration method. A select TMS of a local anesthetic agent (lidocaine or tetracaine) was directly emulsified to prepare an O/W cream and tested for the anesthetic efficacy on intact human skin. While both lidocaine (6%) and tetracaine (4%) creams were highly effective for dermal anesthesia with a similar onset time, the tetracaine cream exhibited a significantly longer duration of action than the lidocaine cream. An accelerated stability study indicated that lidocaine was significantly more stable than tetracaine in the creams. PMID- 12779281 TI - Comparison of shelf-life estimates for a human insulin pharmaceutical preparation using the matrix and full-testing approaches. AB - The design of a stability study is intended to establish a shelf-life based on testing a limited number of batches of a drug product, which is applicable to all future batches of the drug product manufactured under similar circumstances. In the FDA guidelines for stability testing of new drug products, the matrix design is suggested as the alternative statistical design to the full testing approach. In the last few years, the matrix design has received increasing attention from pharmaceutical companies in order to reduce the cost of stability studies. For this, the full and matrix stability test plans were used on three batches of human insulin pharmaceutical preparation stored under a kinetic mean temperature of 22.7 degrees C for 3 years. The statistical method was suggested by Ruberg and Stegeman (Biometrics, 1991, 47, 1059-1069) and Llabres et al. (Int. J. Pharm. 2000, 204, 61-68), and was used for comparing batches, and the maximum allowable difference in slopes between batches was used as criteria for estimating a common shelf-life. The results obtained show the conservative nature of the matrix approach, although the difference between the matrix and full shelf-life was less than 1 month. The effect of the matrix method on shelf-life was also studied. The results showed that the model for estimating the shelf-life varied as a function of the permutation used, after testing for both slope and intercept poolability at alpha = 0.25. The batches should be pooled for both full testing and matrix design since the critical significance level required to obtain a power of 0.80 was achieved when the maximum allowable difference in slopes between batches was 1%/month, according to Ruberg and Stegeman. In contrast, when the proposed method by Llabres et al. was used, the results varied in function of the maximum allowable difference in slopes. PMID- 12779282 TI - Highly stabilized amorphous 3-bis(4-methoxyphenyl)methylene-2-indolinone (TAS 301) in melt-adsorbed products with silicate compounds. AB - 3-Bis(4-Methoxyphenyl)methylene-2-indolinone (TAS-301) is a poorly water-soluble drug showing low oral bioavailability in rats and dogs. Previously, we reported that when a physical mixture of TAS-301 and a porous calcium silicate, Florite RE (FLR), was heated at high temperature (250 degrees C), the drug melted and was adsorbed by the FLR in an amorphous state, and that the preparation (melt adsorbed product) showed a significantly increased solubility and dissolution rate, and a significantly enhanced oral bioavailability of the drug. The aim of the present study was to elucidate important factors for preparing a melt adsorbed product showing greater stability of drug in an amorphous state. We examined the effects of the kind of adsorbent, drug/adsorbent ratio, heating conditions, and drug particle size on converting drug crystal into an amorphous state, the stability of amorphous state, and chemical stability of the drug in the melt-adsorbed products under a high temperature and high humidity condition (60 degrees C/80% RH, open). FLR, light anhydrous silicic acid and two types of hydrated silicon dioxides were tested as adsorbents. For the batch method, TAS 301 was converted into an amorphous state by heating TAS-301/adsorbents physical mixtures above the melting point of TAS-301 for more than 2 min. The amorphous state was most stabilized when FLR was used as an adsorbent and drug/FLR ratio was 1:0.5 and more. For the continuous method using the twin screw extruder that enables significantly larger scale manufacturing than batch method, TAS-301 melt adsorbed products were able to produce when only FLR was used as adsorbent. The heating temperature was needed to be set above the melting point of TAS-301 to convert it into an amorphous state as well as batch method. The amorphous state was stabilized when drug/FLR ratio was 1:2 and more. The micronization of the drug decreased the stability of the amorphous state. These results indicate the importance of optimizing the above factors in the preparation of melt-adsorbed product. PMID- 12779283 TI - Semisolid matrix filled capsules: an approach to improve dissolution stability of phenytoin sodium formulation. AB - Seven semisolid fill bases were selected for the formulation of 24 capsule formulations, each containing 100 mg of phenytoin sodium. The fill materials were selected based on the water absorption capacity of their mixtures with phenytoin sodium. The fill matrices included lipophilic bases (castor oil, soya oil, and Gelucire (G) 33/01), amphiphilic bases (G 44/14 and Suppocire BP), and water soluble bases (PEG 4000 and PEG 6000). The drug:base ratio was 1:2. Excipients such as lecithin, docusate sodium, and poloxamer 188 were added to some formulations. The dissolution rate study indicated that formulations containing lipophilic and amphiphilic bases showed the best release profiles. These are F4 (castor oil-1% docusate sodium); F10 (castor oil-3% poloxamer 188); F14 (G33/01 10% lecithin); F17 (G33/01-1% docusate sodium), and F20 (Suppocire BP). Further, the dissolution stability of the five formulations above was assessed by an accelerated stability study at 30 degrees C and 75% RH using standard Epanutin capsules for comparison. The study included the test and standard capsules either packed in the container of marketed Epanutin capsules (packed) or removed from their outer pack (unpacked). Release data indicated superior release rates of castor oil based formulations (F4 and F10) relative to standard capsules in both the unpacked and packed forms. For instance, the extent of drug release at 30 min after 1 month was 91% for F4 and F10 and 20% for standard capsules. Drug release from packed capsules after 6 months storage was 88% for both formulations F4 and F10 and 35% for standard capsules. In conclusion, the pharmaceutical quality of phenytoin sodium capsules can be improved by using a semisolid lipophilic matrix filled in hard gelatin capsules. PMID- 12779284 TI - Physicochemical characterization of diclofenac sodium-loaded poloxamer gel as a rectal delivery system with fast absorption. AB - Rectal poloxamer gel systems composed of poloxamers and bioadhesive polymers were easy to administer to the anus and were mucoadhesive to the rectal tissues without leakage after the dose. However, a poloxamer gel containing diclofenac sodium could not be developed using bioadhesive polymers, since the drug was precipitated in this preparation. To develop a poloxamer gel using sodium chloride instead of bioadhesive polymers, the physicochemical properties such as gelation temperature, gel strength, and bioadhesive force of various formulations composed of diclofenac sodium, poloxamers, and sodium chloride were investigated. Furthermore, the pharmacokinetic study of diclofenac sodium delivered by the poloxamer gel was performed. Diclofenac sodium significantly increased the gelation temperature and weakened the gel strength and bioadhesive force, while sodium chloride did the opposite. The poloxamer gels with less than 1.0% sodium chloride, in which the drug was not precipitated, were inserted into the rectum without difficulty and leakage, and were retained in the rectum of rats for at least 6 hr. Furthermore, poloxamer gel gave significantly higher initial plasma concentrations and faster Tmax of diclofenac sodium than did solid suppository, indicating that drug from poloxamer gel could be absorbed faster than that from the solid one in rats. Our results suggested that a rectal poloxamer gel system with sodium chloride and poloxamers was a more physically stable, convenient, and effective rectal dosage form for diclofenac sodium. PMID- 12779285 TI - Potential applications of polymeric microsphere suspension as subcutaneous depot for insulin. AB - The objective of this investigation was to develop an injectable, depot-forming drug delivery system for insulin based on microparticle technology to maintain constant plasma drug concentrations over prolonged period of time for the effective control blood sugar levels. Formulations were optimized with two well characterized biodegradable polymers namely, poly(DL-lactide-co-glycolide) and poly-epsilon-caprolactone and evaluated in vitro for physicochemical characteristics, drug release in phosphate buffered saline (pH 7.4), and evaluated in vivo in streptozotocin-induced hypoglycemic rats. With a large volume of internal aqueous phase during w/o/w double emulsion solvent evaporation process and high molecular weight of the polymers used, we could not achieve high drug capture and precise control over subsequent release within the study period of 60 days. However, this investigation revealed that upon subcutaneous injection, the biodegradable depot-forming polymeric microspheres controlled the drug release and plasma sugar levels more efficiently than plain insulin injection. Preliminary pharmacokinetic evaluation exhibited steady plasma insulin concentration during the study period. These formulations, with their reduced frequency of administration and better control over drug disposition, may provide an economic benefit to the user compared with products currently available for diabetes control. PMID- 12779286 TI - Development of modified release diltiazem HCl tablets using composite index to identify optimal formulation. AB - This article reports the preparation of tartaric acid treated ispaghula husk powder for the development of modified release tablets of diltiazem HCl by adopting direct compression technique and a 32 full factorial design. The modified ispaghula husk powder showed superior swelling and gelling as compared to untreated powder. Addition of compaction augmenting agent such as dicalcium phosphate was found to be essential for obtaining tablets with adequate crushing strength. In order to improve the crushing strength of diltiazem HCl tablets, to modulate drug release pattern, and to obtain similarity of dissolution profiles in distilled water and simulated gastric fluid (pH 1.2), modified guar gum was used along with modified ispaghula husk powder and tartaric acid. A novel composite index, which considers a positive or a negative deviation from an ideal value, was calculated considering percentage drug release in 60, 300, and 540 min as dependent variables for the selection of a most appropriate batch. Polynomial equation and contour plots are presented. The concept of similarity factor (f2) was used to prove similarity of dissolution in water and simulated gastric fluid (pH 1.2). PMID- 12779287 TI - Film coated pellets containing verapamil hydrochloride: enhanced dissolution into neutral medium. AB - Weakly basic drugs, such as verapamil hydrochloride, that are poorly soluble in neutral/alkaline medium may have poor oral bioavailability due to reduced solubility in the small intestine and colon. Film coated pellets were prepared using two strategies to enhance drug release at high pH values. Firstly, pellets were coated with Eudragit RS/hydroxypropyl methylcellulose acetate succinate (HMAS) mixtures in proportions of 10:1 and 10:3, respectively. The enteric polymer, HMAS, would dissolve in medium at pH > 6 creating pores through the insoluble Eudragit RS membrane to increase drug release. Secondly, an acidic environment was created within the core by the inclusion of fumaric acid at concentrations of 5 and 10% in order to increase drug solubility. Both strategies enhanced drug release into neutral medium in dissolution studies using the pH change method to simulate GIT transit. Dissolution profiles of samples tested in pH 1.2 for 12 hr were compared with those using the pH change method (pH 1.2 for first 1.5 hr, pH raised to 6.8 for remaining 10.5 hr) using the area under the dissolution curve (AUC), the dissolution half-life (t50%), and the amount of drug released in 3 hr (A3hr) values. Both strategies enhanced drug release into neutral medium although the strategy using HMAS in the film was more effective. The formulation least affected by pH change was a combination of the two strategies, i.e., pellets containing 5% fumaric acid coated with Eudragit RS 12% w/w and HMAS 1.2% w/w. PMID- 12779288 TI - Effect of anionic water-soluble dyes on film coating properties of chitosan acetate. AB - Solution of chitosan in dilute acetic acid was prepared to have an apparent viscosity of 125 mPa s and mixed with solution of anionic water-soluble dye. The effects of concentration and type of dye and molecular weight and percentage deacetylation of chitosan on their miscibility and physical stability were investigated. High concentration of dye and high molecular weight and percentage deacetylation of chitosan resulted in precipitation or colloidal dispersion due to ionic interaction between dye and the polymer. The effect was more prominent upon storage. The miscibility of dye and the polymer depended on the molecular configuration and ionic group in the dye molecule. It was ranked brilliant blue approximately euqal to green FS > fast green > ponceau SX approximately euqal to green sunset yellow > erythrosine approximately euqal to green tartrazin > indigo carmine. Solutions of low molecular weight chitosan with and without green FS were then used as coating formulations onto propranolol hydrochloride core tablets. There was no color migration on coated tablets even after storage for 1 year. Disintegration and drug dissolution from tablets coated with colored film were slightly slower than those from tablets coated with plain film and core tablet, respectively. This was corresponding to the results of swelling and dissolution of cast films. However, all tablets conformed to the specification in monograph of USP XXIV. PMID- 12779289 TI - Lyophilization of unit dose pharmaceutical dosage forms. AB - A lyophilization process for a pharmaceutical unit dosage form was developed which comprised a container closed with an impermeable membrane pierced with one or more holes through which the material in the container can be lyophilized. The hole or holes in the membrane have to be sufficiently large to allow water vapor to escape but small to ensure that the material is kept within the container. Lyophilization from sealed, perforated, unit-dose package has shown to be feasible. The technique offers a novel convenient means of lyophilizing nonsterile products in their primary pack and increases the potential for the development of lyophilized formulations for nonparenteral applications. PMID- 12779290 TI - Postmenopausal health: a reassessment post WHI. PMID- 12779291 TI - Alternatives to HRT: an evidence-based review. AB - In the search for alternatives to HRT--a search motivated by the yearning for "natural" treatments and, more recently, by bad news from HRT trials--herbs and botanicals, and some foods, have received much attention and wide acceptance. This acceptance is, however, based on little more than advertising and consumers' desire to believe it. The available research is mainly not of high quality, and there remains a host of products to be tested. The chief problems have been lack of standards for admission to study, unknown dosages of active principles, and unclear or lax criteria for effectiveness. So far, though, black cohosh and soy (in many forms) can be said to offer some benefits. PMID- 12779292 TI - Unusual late complications after two previous cesarean deliveries: a case report. AB - BACKGROUND: Women with multiple previous cesarean deliveries (CDs) risk the development of uterine synechiae and ventral fixation of the uterus to the abdominal wall. CASE: A para 2, gravida 2 women who had two prior CDs experienced prolonged menstrual bleeding with persistent cramps, both of which became more severe after insertion of an intrauterine contraceptive device that was subsequently removed. Pelvic ultrasound revealed an enlarged uterus with fibroids. After a failed attempt to perform a dilation and curettage because of uterine synechiae, the patient underwent a total abdominal hysterectomy. During surgery, the gynecologist observed a total dehiscence of the previous uterine incision, with ventral fixation of the uterus to the lower anterior abdominal wall and marked elongation of the portio cervix. CONCLUSION: Dysmenorrhea in a patient with multiple previous CDs, mainly of the classical type, should be carefully evaluated and, in addition, when such patient requires a dilation and curettage, the possibility of cervical stenosis and uterine synechiae should be kept in mind. Having had a number of previous CDs may have an adverse impact on the uterine complication rate. PMID- 12779293 TI - Evaluation of an indirect immunofluorescence assay for detecting Chlamydia trachomatis as a method for diagnosing tubal factor infertility in Mexican women. AB - To evaluate the clinical impact of the use of an indirect immunofluorescence assay (IFA) against Chlamydia as a method to identify patients with tubal factor infertility (TFI) in a population of infertile Mexican women. METHODS: This was a retrospective analysis made on 100 patients attending the infertility clinic who underwent laparoscopy. Blood and cervical samples were collected during the clinical examination. The presence of anti-Chlamydia trachomatis IgG antibodies was documented using the IFA test, and the presence of active chlamydial infection was evaluated using the nucleic acid hybridization assay. RESULTS: The sensitivity and specificity values of the IFA test to identify patients with periadnexal adhesions were 45% and 82%, respectively; and the positive predictive and negative predictive values were 42% and 84%, and the positive and negative likelihood ratios were 2.5 and 0.7, respectively. CONCLUSION: The IFA test was not usable for the identification of patients with periadnexal adhesions as a cause of infertility in this population. However, it could be useful as a screening test to decide which patients might receive laparoscopic treatment. Furthermore, it could be useful for identifying patients with active chlamydial infections in the upper genital tract, but a study with a larger sample needs to be done. PMID- 12779294 TI - Do characteristic spermatozoal morphological abnormalities exist in patients who have undergone unilateral orchiectomy and preventive radiotherapy? AB - DESIGN: We examined 16 men who had been subjected to unilateral orchiectomy owing to seminoma and to preventive radiotherapy, in order to investigate the morphologic abnormalities of the spermatozoa (headless and small-round headedness) that may contribute to infertility. RESULTS: The same morphologic abnormalities of the head and neck found in the semen samples of fertile men were also found in the semen samples of the patients, albeit in higher percentages; the morphologic abnormalities of the tail in the semen samples of the patients were similar to those of the fertile men, both qualitatively and quantitatively. CONCLUSION: No specifically characteristic morphologic abnormalities of the spermatozoa were detected in men who were subjected to unilateral orchiectomy and to preventive radiotherapy in comparison with fertile men. The percentage rate of morphologic tail abnormalities is not affected by preventive radiotherapy. PMID- 12779295 TI - Sperm velocity and morphology, female characteristics, and the hypo-osmotic swelling test as predictors of fertilization potential: experience from the IVF model. AB - OBJECTIVE: To determine, among the following variables, the predictors of sperm fertilization potential of a given semen sample: sperm count, eight sperm velocity characteristics determined by computer-assisted semen analysis, the sperm morphology score determined by strict criteria, and the hypo-osmotic swelling (HOS) test, using the IVF model. DESIGN: Data from 58 couples attending the assisted conception unit for conventional IVF treatment were analyzed retrospectively by simple linear regression and multiple stepwise regression analysis, taking the fertilization rate as the dependent factor. RESULTS: The mean sperm velocity and the strict sperm morphology assessment were the only parameters showing significant correlation with the fertilization rate. A minimum sperm velocity of 13 microm/s and a minimum strict sperm morphology of 2% were necessary to achieve fertilization in 50% of the oocytes. The results of the HOS test did not correlate with the fertilization rate. An equation was devised to calculate the expected fertilization rate based on the mean sperm velocity and the strict morphology score, and can be used to counsel patients prior to assisted conception. CONCLUSIONS: The mean sperm velocity and strict sperm morphology evaluation are good predictors of fertilization potential, but the HOS test is not. PMID- 12779296 TI - Which patients with acute exacerbation of chronic obstructive pulmonary disease benefit from noninvasive positive-pressure ventilation? A systematic review of the literature. AB - BACKGROUND: Over the past decade, noninvasive positive-pressure ventilation (NPPV) in the setting of acute exacerbations of chronic obstructive pulmonary disease (COPD) has increased in popularity. Although several trials have been published on the relative effectiveness of this treatment, apparent inconsistencies in study results remain. PURPOSE: To assess the effect of NPPV on rate of endotracheal intubation, length of hospital stay, and in-hospital mortality rate in patients with an acute exacerbation of COPD and to determine the effect of exacerbation severity on these outcomes. DATA SOURCES: MEDLINE (1966 to 2002) and EMBASE (1990 to 2002). Additional data sources included the Cochrane Library, personal files, abstract proceedings, reference lists of selected articles, and expert contact. There were no language restrictions. STUDY SELECTION: The researchers selected randomized, controlled trials that 1) examined patients with acute exacerbation of COPD; 2) compared noninvasive ventilation and standard therapy with standard therapy alone; and 3) included need for endotracheal intubation, length of hospital stay, or hospital survival as an outcome. DATA EXTRACTION: Methodologic quality and results were abstracted independently and in duplicate. DATA SYNTHESIS: The addition of NPPV to standard care in patients with an acute exacerbation of COPD decreased the rate of endotracheal intubation (risk reduction, 28% [95% CI, 15% to 40%]), length of hospital stay (absolute reduction, 4.57 days [CI, 2.30 to 6.83 days]), and in hospital mortality rate (risk reduction, 10% [CI, 5% to 15%]). However, subgroup analysis showed that these beneficial effects occurred only in patients with severe exacerbations, not in those with milder exacerbations. CONCLUSIONS: Patients with severe exacerbations of COPD benefit from the addition of NPPV to standard therapy. However, NPPV has not been shown to benefit hospitalized patients with milder COPD exacerbations. PMID- 12779297 TI - Spinal manipulative therapy for low back pain. A meta-analysis of effectiveness relative to other therapies. AB - BACKGROUND: Low back pain is a costly illness for which spinal manipulative therapy is commonly recommended. Previous systematic reviews and practice guidelines have reached discordant results on the effectiveness of this therapy for low back pain. PURPOSE: To resolve the discrepancies related to use of spinal manipulative therapy and to update previous estimates of effectiveness by comparing spinal manipulative therapy with other therapies and then incorporating data from recent high-quality randomized, controlled trials (RCTs) into the analysis. DATA SOURCES: MEDLINE, EMBASE, CINAHL, the Cochrane Controlled Trials Register, and previous systematic reviews. STUDY SELECTION: Randomized, controlled trials of patients with low back pain that evaluated spinal manipulative therapy with at least 1 day of follow-up and at least one clinically relevant outcome measure. DATA EXTRACTION: Two authors, who served as the reviewers for all stages of the meta-analysis, independently extracted data from unmasked articles. Comparison treatments were classified into the following seven categories: sham, conventional general practitioner care, analgesics, physical therapy, exercises, back school, or a collection of therapies judged to be ineffective or even harmful (traction, corset, bed rest, home care, topical gel, no treatment, diathermy, and minimal massage). DATA SYNTHESIS: Thirty-nine RCTs were identified. Meta-regression models were developed for acute or chronic pain and short-term and long-term pain and function. For patients with acute low back pain, spinal manipulative therapy was superior only to sham therapy (10-mm difference [95% CI, 2 to 17 mm] on a 100-mm visual analogue scale) or therapies judged to be ineffective or even harmful. Spinal manipulative therapy had no statistically or clinically significant advantage over general practitioner care, analgesics, physical therapy, exercises, or back school. Results for patients with chronic low back pain were similar. Radiation of pain, study quality, profession of manipulator, and use of manipulation alone or in combination with other therapies did not affect these results. CONCLUSIONS: There is no evidence that spinal manipulative therapy is superior to other standard treatments for patients with acute or chronic low back pain. PMID- 12779298 TI - Accepting critically ill transfer patients: adverse effect on a referral center's outcome and benchmark measures. AB - BACKGROUND: Common methods of benchmarking clinical performance rarely, if ever, account for admission source and, in particular, the effect of a patient being transferred from one medical center to another. Small biases in comparisons of observed versus expected deaths can substantially affect how high-quality institutions compare with peer hospitals. With the most sophisticated and validated set of case-mix measures available for patients, the intensive care unit is an ideal setting in which to study the effect of a patient's being transferred from another hospital. OBJECTIVE: To determine the extent of bias in benchmarking outcomes when performance measures do not account for transfer patients' greater severity of illness. DESIGN: Prospectively developed cohort study. SETTING: Medical intensive care unit (MICU) at a tertiary care university hospital. PATIENTS: 4579 consecutive admissions for 4208 patients from 1 January 1994 to 1 April 1998. MEASUREMENTS: MICU and hospital lengths of stay, MICU readmission, and hospital mortality rates. RESULTS: Compared with directly admitted patients, MICU patients transferred from another hospital had significantly higher Acute Physiology Scores at the time of admission and discharge (P = 0.001). Even after full adjustment for case mix and severity of illness, transfer patients had a 38% longer MICU stay (95% CI, 32% to 45%), a 41% longer hospital stay (CI, 34% to 50%), and a 2.2 times greater odds of hospital mortality (CI, 1.7 to 2.8) than directly admitted patients. With identical efficiency and quality, a referral hospital with a 25% MICU transfer rate compared with another with a 0% transfer rate would be penalized by 14 excess deaths per 1000 admissions when a benchmarking program adjusts only for case mix and severity of illness and not for the source of admission. CONCLUSIONS: In a setting with the most thorough diagnostic-based, case-mix adjustment and the most physiologically precise severity-of-illness information, accepting transfer patients can adversely affect efficiency and quality benchmarks. Benchmarking and profiling efforts beyond intensive care units must also recognize and account for this phenomenon; otherwise, referral centers may have an incentive to refuse care for patients who could benefit from being transferred to their facility. PMID- 12779299 TI - Relationship between cigarette smoking and novel risk factors for cardiovascular disease in the United States. AB - BACKGROUND: Few studies have examined the relationship between cigarette smoking and novel risk factors for cardiovascular disease in a general population or have included a biochemical marker of current smoking. OBJECTIVE: To examine the relationship between cigarette smoking and serum C-reactive protein, fibrinogen, and homocysteine levels. DESIGN: Cross-sectional study. SETTING: The U.S. general population. PATIENTS: 4187 current smokers, 4791 former smokers, and 8375 never smokers 18 years of age or older who participated in the Third National Health and Nutrition Examination Survey conducted between 1988 and 1994. MEASUREMENTS: Serum C-reactive protein levels were categorized as detectable (2.2 to 9.9 mg/L) or clinically elevated (> or =10 mg/L), and fibrinogen and homocysteine levels were defined as elevated if in the 85th percentile or greater (11.1 micromol/L and 12.7 mmol/L, respectively). RESULTS: After adjustment for traditional cardiovascular disease risk factors, cigarette smoking was related to elevated levels of C-reactive protein, fibrinogen, and homocysteine. Compared with never smoking cigarettes, self-reported current cigarette smoking was associated with a C-reactive protein level in the detectable (odds ratio, 1.66 [95% CI, 1.40 to 1.97]; P < 0.001) or clinically elevated (odds ratio, 1.98 [CI, 1.57 to 2.51]; P < 0.001) ranges, with elevated levels of fibrinogen (odds ratio, 2.15 [CI, 1.65 to 2.80]; P < 0.001) and homocysteine (odds ratio, 2.10 [CI, 1.62 to 2.74]; P < 0.001). There were positive and significant dose-response relationships between measures of cigarette smoking (cigarettes per day, pack-years, and serum cotinine levels) and elevated levels of novel risk factors. CONCLUSIONS: These findings suggest that inflammation and hyperhomocysteinemia may be important mechanisms by which smoking promotes atherosclerotic disease. PMID- 12779300 TI - A review of the evidence for the effectiveness, safety, and cost of acupuncture, massage therapy, and spinal manipulation for back pain. AB - BACKGROUND: Few treatments for back pain are supported by strong scientific evidence. Conventional treatments, although widely used, have had limited success. Dissatisfied patients have, therefore, turned to complementary and alternative medical therapies and providers for care for back pain. PURPOSE: To provide a rigorous and balanced summary of the best available evidence about the effectiveness, safety, and costs of the most popular complementary and alternative medical therapies used to treat back pain. DATA SOURCES: MEDLINE, EMBASE, and the Cochrane Controlled Trials Register. STUDY SELECTION: Systematic reviews of randomized, controlled trials (RCTs) that were published since 1995 and that evaluated acupuncture, massage therapy, or spinal manipulation for nonspecific back pain and RCTs published since the reviews were conducted. DATA EXTRACTION: Two authors independently extracted data from the reviews (including number of RCTs, type of back pain, quality assessment, and conclusions) and original articles (including type of pain, comparison treatments, sample size, outcomes, follow-up intervals, loss to follow-up, and authors' conclusions). DATA SYNTHESIS: Because the quality of the 20 RCTs that evaluated acupuncture was generally poor, the effectiveness of acupuncture for treating acute or chronic back pain is unclear. The three RCTs that evaluated massage reported that this therapy is effective for subacute and chronic back pain. A meta-regression analysis of the results of 26 RCTs evaluating spinal manipulation for acute and chronic back pain reported that spinal manipulation was superior to sham therapies and therapies judged to have no evidence of a benefit but was not superior to effective conventional treatments. CONCLUSIONS: Initial studies have found massage to be effective for persistent back pain. Spinal manipulation has small clinical benefits that are equivalent to those of other commonly used therapies. The effectiveness of acupuncture remains unclear. All of these treatments seem to be relatively safe. Preliminary evidence suggests that massage, but not acupuncture or spinal manipulation, may reduce the costs of care after an initial course of therapy. PMID- 12779301 TI - The epidemiology of "asymptomatic" left ventricular systolic dysfunction: implications for screening. AB - Congestive heart failure is a progressive disorder that is frequently preceded by asymptomatic left ventricular systolic dysfunction. We reviewed the epidemiology, diagnosis, and natural history of asymptomatic left ventricular systolic dysfunction and evaluated community-wide screening for this condition as a potential strategy to reduce the incidence of heart failure. Asymptomatic left ventricular systolic dysfunction has an estimated prevalence of 3% to 6%, and is at least as common in the community as systolic heart failure. Because it often occurs in the absence of known cardiovascular disease, this condition may go unrecognized and undertreated. In randomized trials, individuals with asymptomatic left ventricular systolic dysfunction have high rates of incident heart failure and death. However, little is known about the prognosis of individuals with this condition in the community, who have a substantially lower prevalence of myocardial infarction, have milder degrees of systolic dysfunction, and are older than patients enrolled in clinical trials. Current evidence is inadequate to support community-wide screening for asymptomatic left ventricular systolic dysfunction, either with echocardiography or with assays for natriuretic peptides. Given the increasing prevalence of heart failure, additional studies are needed to develop effective strategies to detect and optimally manage individuals with asymptomatic left ventricular dysfunction in the community. PMID- 12779302 TI - Pharmacotherapy for heart failure in patients with renal insufficiency. AB - Clinical trials have demonstrated that angiotensin-converting enzyme (ACE) inhibitors, beta-blockers, and spironolactone improve survival in patients with heart failure. Because patients with heart failure and renal insufficiency have been underrepresented in these trials, little evidence is available to guide clinicians in the optimal management of patients with both conditions. Approximately one third to one half of patients with heart failure have renal insufficiency (estimated glomerular filtration rate [GFR] <60 mL/min per 1.73 m2), and renal insufficiency is among the strongest predictors of mortality in patients with heart failure. Evidence supports the use of ACE inhibitors to improve survival in patients with moderate renal insufficiency (GFR, 30 to 60 mL/min per 1.73 m2), but there is little evidence with which to weigh the risks and benefits in patients with more advanced renal dysfunction. beta-Blockers improve survival in patients with heart failure, and their beneficial effect is unlikely to differ according to renal function. Spironolactone improves outcomes in patients with advanced heart failure, but renal insufficiency appears to increase risk for hyperkalemia and limits the use of the drug in patients with severe renal insufficiency. Future clinical trials in heart failure should include a representative number of patients with renal insufficiency to improve the evidence base and outcomes in this vulnerable population. PMID- 12779303 TI - Screening for dementia: recommendation and rationale. PMID- 12779304 TI - Screening for dementia in primary care: a summary of the evidence for the U.S. Preventive Services Task Force. AB - BACKGROUND: Dementia is a large and growing problem but is often not diagnosed in its earlier stages. Screening and earlier treatment could reduce the burden of suffering of this syndrome. PURPOSE: To review the evidence of benefits and harms of screening for and earlier treatment of dementia. DATA SOURCES: MEDLINE, PsycINFO, EMBASE, the Cochrane Library, experts, and bibliographies of reviews. STUDY SELECTION: The authors developed eight key questions representing a logical chain between screening and improved health outcomes, along with eligibility criteria for admissible evidence for each question. Admissible evidence was obtained by searching the data sources. DATA EXTRACTION: Two reviewers abstracted relevant information using standardized abstraction forms and graded article quality according to U.S. Preventive Services Task Force criteria. DATA SYNTHESIS: No randomized, controlled trial of screening for dementia has been completed. Brief screening tools can detect some persons with early dementia (positive predictive value < or =50%). Six to 12 months of treatment with cholinesterase inhibitors modestly slows the decline of cognitive and global clinical change scores in some patients with mild to moderate Alzheimer disease. Function is minimally affected, and fewer than 20% of patients stop taking cholinesterase inhibitors because of side effects. Only limited evidence indicates that any other pharmacologic or nonpharmacologic intervention slows decline in persons with early dementia. Although intensive multicomponent caregiver interventions may delay nursing home placement of patients who have caregivers, the relevance of this finding for persons who do not yet have caregivers is uncertain. Other potential benefits and harms of screening have not been studied. CONCLUSIONS: Screening tests can detect undiagnosed dementia. In persons with mild to moderate clinically detected Alzheimer disease, cholinesterase inhibitors are somewhat effective in slowing cognitive decline. The effect of cholinesterase inhibitors or other treatments on persons with dementia detected by screening is uncertain. PMID- 12779305 TI - Applying evidence to patient care: from black and white to shades of grey. PMID- 12779308 TI - Ethics and complementary and alternative medicine. PMID- 12779309 TI - Summaries for patients. Noninvasive positive-pressure ventilation for severe worsening of chronic obstructive pulmonary disease. PMID- 12779310 TI - Summaries for patients. The effectiveness of spinal manipulation relative to other therapies for low back pain. PMID- 12779311 TI - Summaries for patients. Accepting critically ill transfer patients. PMID- 12779312 TI - Summaries for patients. Associations between smoking and newer risk factors for cardiovascular disease. PMID- 12779313 TI - Summaries for patients. Screening for asymptomatic heart failure. PMID- 12779314 TI - Summaries for patients. Screening for dementia in primary care settings. PMID- 12779315 TI - Urinary catheters: a one-point restraint? PMID- 12779316 TI - A lesson in poverty. PMID- 12779317 TI - New phases of phospholipids and implications to the membrane fusion problem. AB - Membrane fusion is a ubiquitous process in eukaryotic cells. When two membranes fuse, lipid must undergo molecular rearrangements at the point of merging. To understand how lipid structure transitions occur, scientists studied the phase transition of lipid between the lamellar (L(alpha)) phase and the inverted hexagonal (H(II)) phase, based on the idea that lipid must undergo a similar rearrangement as in fusion. However, previous investigations on the system of dioleoylphosphatidylcholine (DOPC) and dioleoylphosphatidylethanolamine (DOPE) did not reveal intermediate phases between the L(alpha) and H(II) phases. Recently, we found a rhombohedral phase of diphytanoylphosphatidylcholine between its L(alpha) and H(II) phases using substrate-supported samples. Here we report the observation of two new phases in the DOPC-DOPE system: a rhombohedral phase and a distorted hexagonal phase. The rhombohedral phase confirms the stalk hypothesis for the L(alpha)-H(II) transition, but the phase of stable stalks exists only for a certain range of spontaneous curvature. The distorted hexagonal phase exists only in a lipid mixture. It implies that lipids may demix to adjust its local spontaneous curvature in order to achieve energy minimum under stress. PMID- 12779318 TI - Both aromatic and cationic residues contribute to the membrane-lytic and bactericidal activity of eosinophil cationic protein. AB - Eosinophil cationic protein (ECP) and eosinophil derived neurotoxin (EDN) are proteins of the ribonuclease A (RNase A) superfamily that have developed biological properties related to the function of eosinophils. ECP is a potent cytotoxic molecule, and although the mechanism is still unknown this cytotoxic activity has been associated with its highly cationic character. Using liposome vesicles as a model, we have demonstrated that ECP tends to disrupt preferentially acidic membranes. On the basis of structure analysis, ECP variants modified at basic and hydrophobic residues have been constructed. Changes in the leakage of liposome vesicles by these ECP variants have indicated the role of both aromatic and basic specific amino acids in cellular membrane disruption. This is the case with the two tryptophans at positions 10 and 35, but not phenylalanine 76, and the two arginines 101 and 104. The bactericidal activity of both native ECP and point-mutated variants, tested against Escherichia coli and Staphylococcus aureus, suggests that basic amino acids play, in addition to the effect on the disruption of the cellular membrane, other roles such as specific binding on the surface of the bacteria cell. PMID- 12779319 TI - Structural features of paramyxovirus F protein required for fusion initiation. AB - On the basis of the coordinates of the related Newcastle disease virus (NDV) F protein, Valine-94, a determinant of measles virus (MV) cytopathicity, is predicted to lie in a cylindrical cavity with 10 A diameter located at the F neck. A 16-residue domain around V94 is functionally interchangeable between NDV and MV F, supporting our homology model. Features of the cavity are conserved within the Paramyxovirinae. A hydrophobic base and a hydrophilic residue at the rim are required for surface expression. Small residue substitutions predicted to open the cavity were found to disrupt transport or limit fusogenicity of transport-competent mutants but can be compensated for by simultaneous insertion of larger residues at the opposing wall. Variants containing histidine substitutions mediate fusion at pH 8.5, while at pH 7.2 fusion is blocked, suggesting that functionality requires low charge in the cavity. These results indicate that specific structural features of the cavity are essential for paramyxovirus fusion initiation. PMID- 12779320 TI - D18G transthyretin is monomeric, aggregation prone, and not detectable in plasma and cerebrospinal fluid: a prescription for central nervous system amyloidosis? AB - Over 70 transthyretin (TTR) mutations facilitate amyloidosis in tissues other than the central nervous system (CNS). In contrast, the D18G TTR mutation in individuals of Hungarian descent leads to CNS amyloidosis. D18G forms inclusion bodies in Escherichia coli, unlike the other disease-associated TTR variants overexpressed to date. Denaturation and reconstitution of D18G from inclusion bodies afford a folded monomer that is destabilized by 3.1 kcal/mol relative to an engineered monomeric version of WT TTR. Since TTR tetramer dissociation is typically rate limiting for amyloid formation, the monomeric nature of D18G renders its amyloid formation rate 1000-fold faster than WT. It is perplexing that D18G does not lead to severe early onset systemic amyloidosis, given that it is the most destabilized TTR variant characterized to date, more so than variants exhibiting onset in the second decade. Instead, CNS impairment is observed in the fifth decade as the sole pathological manifestation; however, benign systemic deposition is also observed. Analysis of heterozygote D18G patient's serum and cerebrospinal fluid (CSF) detects only WT TTR, indicating that D18G is either rapidly degraded postsecretion or degraded within the cell prior to secretion, consistent with its inability to form hybrid tetramers with WT TTR. The nondetectable levels of D18G TTR in human plasma explain the absence of an early onset systemic disease. CNS disease may result owing to the sensitivity of the CNS to lower levels of D18G aggregate. Alternatively, or in addition, we speculate that a fraction of D18G made by the choroid plexus can be transiently tetramerized by the locally high thyroxine (T(4)) concentration, chaperoning it out into the CSF where it undergoes dissociation and amyloidogenesis due to the low T(4) CSF concentration. Selected small molecule tetramer stabilizers can transform D18G from a monomeric aggregation-prone state to a nonamyloidogenic tetramer, which may prove to be a useful therapeutic strategy against TTR associated CNS amyloidosis. PMID- 12779321 TI - gamma-Secretase cleavage and binding to FE65 regulate the nuclear translocation of the intracellular C-terminal domain (ICD) of the APP family of proteins. AB - Regulated intramembrane proteolysis (RIP) of the amyloid precursor protein (APP) produces amyloid beta-protein (Abeta), the probable causative agent of Alzheimer's disease (AD), and is therefore an important target for therapeutic intervention. However, there is a burgeoning consensus that gamma-secretase, one of the proteases that generates Abeta, is also critical for the signal transduction of APP and a growing list of other receptors. APP is a member of a gene family that includes two amyloid precursor-like proteins, APLP1 and APLP2. Although APP and the APLPs undergo similar proteolytic processing, there is little information about the role of their gamma-secretase-generated intracellular domains (ICDs). Here, we show that APLP1 and 2 undergo presenilin dependent RIP similar to APP, resulting in the release of a approximately 6 kDa ICD for each protein. Each of the ICDs are degraded by an insulin degrading enzyme-like activity, but they can be stabilized by members of the FE65 family and translocate to the nucleus. Given that modulation of APP processing is a therapeutic target and that the APLPs are processed in a manner similar to APP, any strategy aimed at altering APP proteolysis will have to take into account possible effects on signaling by APLP 1 and 2. PMID- 12779322 TI - Identification of small-molecule inhibitors of human angiogenin and characterization of their binding interactions guided by computational docking. AB - Angiogenin (ANG) is a potent inducer of angiogenesis and an RNase A homologue whose ribonucleolytic activity is essential for its biological action. Recently, we reported the identification of small non-nucleotide inhibitors of the enzymatic activity of ANG by high-throughput screening (HTS) [Kao, R. Y. T., et al. (2002) Proc. Natl. Acad. Sci. U.S.A. 99, 10066-10071]. Two of the inhibitors that were obtained, National Cancer Institute compound NSC-65828 [8-amino-5-(4' hydroxybiphenyl-4-ylazo)naphthalene-2-sulfonate] and ChemBridge compound C-181431 [4,4'-dicarboxy-3,3'-bis(naphthylamido)diphenylmethanone], were judged to be suitable for further development, and one of these (NSC-65828) was shown to possess antitumor activity in mice. Here we have used computational docking as a guide for the identification of available NSC-65828 and C-181431 analogues that bind more tightly to ANG, and for the characterization of inhibitor binding modes. Numerous analogues were found to have greater avidity than the HTS compounds or any small nucleotide inhibitors; four were considered to be of interest as potential leads (K(i) = 5-25 microM). Two of these analogues bind more tightly to ANG than to RNase A, and are the first small molecules shown to exhibit this selectivity. The predicted binding orientations of the HTS compounds and the new lead inhibitors were evaluated by determining the effects of ANG active site mutations on inhibitory potency. The results with ANG variants R5A, H8A, N68A, and des(121-123) are highly consistent with the docking models. Affinity changes observed with Q12A and Q117G reveal aspects of active site function that are not apparent from the free ANG crystal structure or from the modeled complexes. These findings should prove to be useful in the design of more effective and specific ANG antagonists. PMID- 12779323 TI - Linearization of a naturally occurring circular protein maintains structure but eliminates hemolytic activity. AB - Cyclotides are a recently discovered family of disulfide rich proteins from plants that contain a circular protein backbone. They are exceptionally stable, as exemplified by their use in native medicine of the prototypic cyclotide kalata B1. The peptide retains uterotonic activity after the plant from which it is derived is boiled to make a medicinal tea. The circular backbone is thought to be in part responsible for the stability of the cyclotides, and to investigate its role in determining structure and biological activity, an acyclic derivative, des (24-28)-kalata B1, was chemically synthesized and purified. This derivative has five residues removed from the 29-amino acid circular backbone of kalata B1 in a loop region corresponding to a processing site in the biosynthetic precursor protein. Two-dimensional NMR spectra of the peptide were recorded, assigned, and used to identify a series of distance, angle, and hydrogen bonding restraints. These were in turn used to determine a representative family of solution structures. Of particular interest was a determination of the structural similarities and differences between des-(24-28)-kalata B1 and native kalata B1. Although the overall three-dimensional fold remains very similar to that of the native circular protein, removal of residues 24-28 of kalata B1 causes disruption of some structural features that are important to the overall stability. Furthermore, loss of hemolytic activity is associated with backbone truncation and linearization. PMID- 12779324 TI - The crystal structure, mutagenesis, and activity studies reveal that patatin is a lipid acyl hydrolase with a Ser-Asp catalytic dyad. AB - Patatin is a nonspecific lipid acyl hydrolase that accounts for approximately 40% of the total soluble protein in mature potato tubers, and it has potent insecticidal activity against the corn rootworm. We determined the X-ray crystal structure of a His-tagged variant of an isozyme of patatin, Pat17, to 2.2 A resolution, employing SeMet multiwavelength anomalous dispersion (MAD) phasing methods. The patatin crystal structure has three molecules in the asymmetric unit, an R-factor of 22.0%, and an R(free) of 27.2% (for 10% of the data not included in the refinement) and includes 498 water molecules. The structure notably revealed that patatin has a Ser-Asp catalytic dyad and an active site like that of human cytosolic phospholipase A(2) (cPLA(2)) [Dessen, A., et al. (1999) Cell 97, 349-360]. In addition, patatin has a folding topology related to that of the catalytic domain of cPLA(2) and unlike the canonical alpha/beta hydrolase fold. The structure confirms our site-directed mutagenesis and bioactivity data that initially suggested patatin possessed a Ser-Asp catalytic dyad. Alanine-scanning mutagenesis revealed that Ser77 and Asp215 were critical for both esterase and bioactivity, consistent with prior work implicating a Ser residue [Strickland, J. H., et al. (1995) Plant Physiol. 109, 667-674] and a Ser Asp dyad [Hirschberg, H. J. H. B., et al. (2001) Eur. J. Biochem. 268, 5037-5044] in patatin's catalytic activity. The crystal structure aids the understanding of other structure-function relationships in patatin. Patatin does not display interfacial activation, a hallmark feature of lipases, and this is likely due to the fact that it lacks a flexible lid that can shield the active site. PMID- 12779325 TI - 2.4 A resolution crystal structure of the prototypical hormone-processing protease Kex2 in complex with an Ala-Lys-Arg boronic acid inhibitor. AB - This paper reports the first structure of a member of the Kex2/furin family of eukaryotic pro-protein processing proteases, which cleave sites consisting of pairs or clusters of basic residues. Reported is the 2.4 A resolution crystal structure of the two-domain protein ssKex2 in complex with an Ac-Ala-Lys-boroArg inhibitor (R = 20.9%, R(free) = 24.5%). The Kex2 proteolytic domain is similar in its global fold to the subtilisin-like superfamily of degradative proteases. Analysis of the complex provides a structural basis for the extreme selectivity of this enzyme family that has evolved from a nonspecific subtilisin-like ancestor. The P-domain of ssKex2 has a novel jelly roll like fold consisting of nine beta strands and may potentially be involved, along with the buried Ca(2+) ion, in creating the highly determined binding site for P(1) arginine. PMID- 12779326 TI - New substrates for beta-lactam-recognizing enzymes: aryl malonamates. AB - Aryl malonamates are demonstrated to be novel substrates of a broad range of beta lactam-recognizing enzymes. These compounds are isomers of the aryl phenaceturates, which are well-known substrates of these enzymes, but the new compounds contain a retro-amide side chain. Several lines of evidence, including comparisons of steady-state kinetic parameters between enzymes and a detailed investigation of the methanolysis kinetics, solvent deuterium isotope effects, and pH-rate profile for turnover of a retro substrate by the Enterobacter cloacae P99 beta-lactamase, suggested that the new substrates are likely to be hydrolyzed by the same chemical mechanisms as "normal" substrates. Molecular modeling indicated that the retro-amide group fits snugly into the active site of the P99 beta-lactamase by hydrogen bonding to the conserved lysine-67 residue. The retro amide side chain may represent a lead to novel mechanism-based and transition state analogue inhibitors. PMID- 12779327 TI - Spectroscopic and mutational analysis of the blue-light photoreceptor AppA: a novel photocycle involving flavin stacking with an aromatic amino acid. AB - The flavoprotein AppA is a blue-light photoreceptor that functions as an antirepressor of photosynthesis gene expression in the purple bacterium Rhodobacter sphaeroides. Heterologous expression studies show that FAD binds to a 156 amino acid N-terminal domain of AppA and that this domain is itself photoactive. A pulse of white light causes FAD absorption to be red shifted in a biphasic process with a fast phase occurring in <1 micros and a slow phase occurring at approximately 5 ms. The absorbance shift was spontaneously restored over a 30 min period, also in a biphasic process as assayed by fluorescence quenching and electronic absorption analyses. Site-directed replacement of Tyr21 with Leu or Phe abolished the photochemical reaction implicating involvement of Tyr21 in the photocycle. Nuclear magnetic resonance analysis of wild-type and mutant proteins also indicates that Tyr21 forms pi-pi stacking interactions with the isoalloxazine ring of FAD. We propose that photochemical excitation of the flavin results in strengthening of a hydrogen bond between the flavin and Tyr 21 leading to a stable local conformational change in AppA. PMID- 12779328 TI - A new strategy for the site-specific modification of proteins in vivo. AB - We recently developed a method for genetically incorporating unnatural amino acids site-specifically into proteins expressed in Escherichia coli in response to the amber nonsense codon. Here we describe the selection of an orthogonal tRNA TyrRS pair that selectively and efficiently incorporates m-acetyl-l-phenylalanine into proteins in E. coli. We demonstrate that proteins containing m-acetyl-l phenylalanine or p-acetyl-l-phenylalanine can be selectively labeled with hydrazide derivatives not only in vitro but also in living cells. The labeling reactions are selective and in general proceed with yields of >75%. In specific examples, m-acetyl-l-phenylalanine was substituted for Lys7 of the cytoplasmic protein Z domain, and for Arg200 of the outer membrane protein LamB, and the mutant proteins were selectively labeled with a series of fluorescent dyes. The genetic incorporation of a nonproteinogenic "ketone handle" into proteins provides a powerful tool for the introduction of biophysical probes for the structural and functional analysis of proteins in vitro or in vivo. PMID- 12779329 TI - Nucleotide excision repair from site-specifically platinum-modified nucleosomes. AB - Nucleotide excision repair is a major cellular defense mechanism against the toxic effects of the anticancer drug cisplatin and other platinum-based chemotherapeutic agents. In this study, mononucleosomes were prepared containing either a site-specific cis-diammineplatinum(II)-DNA intrastrand d(GpG) or a d(GpTpG) cross-link. The ability of the histone core to modulate the excision of these defined platinum adducts was investigated as a model for exploring the cellular response to platinum-DNA adducts in chromatin. Comparison of the extent of repair by mammalian cell extracts of free and nucleosomal DNA containing the same platinum-DNA adduct reveals that the nucleosome significantly inhibits nucleotide excision repair. With the GTG-Pt DNA substrate, the nucleosome inhibits excision to about 10% of the level observed with free DNA, whereas with the less efficient GG-Pt DNA substrate the nucleosome inhibited excision to about 30% of the level observed with free DNA. The effects of post-translational modification of histones on excision of platinum damage from nucleosomes were investigated by comparing native and recombinant nucleosomes containing the same intrastrand d(GpTpG) cross-link. Excision from native nucleosomal DNA is approximately 2-fold higher than the level observed with recombinant material. This result reveals that post-translational modification of histones can modulate nucleotide excision repair from damaged chromatin. The in vitro system established in this study will facilitate the investigation of platinum-DNA damage by DNA repair processes and help elucidate the role of specific post translational modification in NER of platinum-DNA adducts at the physiologically relevant nucleosome level. PMID- 12779330 TI - Major kinetic traps for the oxidative folding of leech carboxypeptidase inhibitor. AB - The leech carboxypeptidase inhibitor (LCI) is a 66-amino acid protein, containing four disulfides that stabilize its structure. This polypeptide represents an excellent model for the study and understanding of the diversity of folding pathways in small, cysteine-rich proteins. The pathway of oxidative folding of LCI has been elucidated in this work, using structural and kinetic analysis of the folding intermediates trapped by acid quenching. Reduced and denatured LCI refolds through a rapid, sequential flow of one- and two-disulfide intermediates and reaches a rate-limiting step in which a mixture of three major three disulfide species and a heterogeneous population of non-native four-disulfide (scrambled) isomers coexist. The three three-disulfide intermediates have been identified as major kinetic traps along the folding pathway of LCI, and their disulfide structures have been elucidated in this work. Two of them contain only native disulfide pairings, and one contains one native and two non-native disulfide bonds. The coexistence of three-disulfide kinetic traps adopting native disulfide bonds together with a significant proportion of fully oxidized scrambled isomers shows that the folding pathway of LCI features properties exhibited by both the bovine pancreatic trypsin inhibitor and hirudin, two diverse models with extreme folding characteristics. The results further demonstrate the large diversity of disulfide folding pathways. PMID- 12779332 TI - Monitoring RNA base structure and dynamics using site-directed spin labeling. AB - Site-directed spin labeling utilizes site-specific attachment of a stable nitroxide radical to probe the structure and dynamics of macromolecules. In the present study, a 4-thiouridine base is introduced at each of six different positions in a 23-nucleotide RNA molecule. The 4-thiouridine derivatives were subsequently modified with one of three methanethiosulfonate nitroxide reagents to introduce a spin label at specific sites. The electron paramagnetic resonance spectra of the labeled RNAs were analyzed in terms of nitroxide motion and the RNA solution structure. At a base-paired site in the RNA helix, where the nitroxide has weak or no local interactions, motion of the nitroxide is apparently dominated by rotation about bonds within the probe. The motion is similar to that found for a structurally related probe on helical sites in proteins, suggesting a similar mode of motion. At other sites that are hydrogen bonded and stacked within the helix, local interactions within the RNA molecule modulate the nitroxide motion in a manner consistent with expectations based on the known structure. For a base that is not structurally constrained, the mobility is higher than at any other site, presumably due to motion of the base itself. These results demonstrate the general utility of the 4 thiouridine/methanethiosulfonate coupling method to introduce nitroxide spin labels into RNA and the ability of the resulting label to probe local structure and dynamics. PMID- 12779331 TI - Regulation and mutational analysis of the HPr kinase/phosphorylase from Bacillus subtilis. AB - In most Gram-positive bacteria, catabolite repression is mediated by a bifunctional enzyme, the HPr kinase/phosphorylase (HprK/P). It has recently been shown that HprK/P could catalyze the phosphorylation of the protein HPr by using pyrophosphate (PP(i)) as a phosphate donor instead of ATP. Here we showed that, as for ATP, PP(i) binds to the enzyme with strong positive cooperativity. However, in contrast to ATP, PP(i) binding does not modify the fluorescence properties of the unique Trp residue of Bacillus subtilis HprK/P. In addition, to understand how two conserved motifs, namely, the P-loop and the specific signature of this family, participate in the three enzymatic activities of HprK/Ps (ATP-kinase, PP(i)-kinase, and phosphorylase), several site-directed mutants were generated. Whereas the three activities are mediated by the P-loop which is directly involved in the binding of ATP, PP(i), or Pi, the signature motif seems to be involved preferentially in the dephosphorylation reaction. On the basis of these results, we propose a model in which the binding of the allosteric activator FBP induces a conformational change of a central loop located above the active site of HprK/P, thereby allowing the ATP binding. However, this conformational change is not required for the binding of PP(i). PMID- 12779333 TI - A second site rescue mutation partially restores functional expression to the serotonin transporter mutant V382P. AB - Transmembrane span 7 (TM7) of the serotonin transporter (SERT) was previously subjected to random mutagenesis, and the mutation V382P was found to abolish transport activity. Val-382 lies next to a threonine residue in the native sequence, creating a TP motif in this mutant. On the basis of molecular modeling studies, which have shown that the presence of a TP motif produces a very large kink in an alpha-helix, it was hypothesized that this motif could be the source of V382P's deleterious effects. We tested this hypothesis by producing second site mutations in the V382P construct that removed the TP motif: T381A-V382P and T381V-V382P. These mutants were tested for the recovery of serotonin transport and binding activities and for expression at the cell surface. The TM7 alpha helix was modeled computationally, using Biased Monte Carlo simulations to quantify the conformational preferences of the wild type and mutant helices. The double mutation T381A-V382P, which was predicted by modeling to produce a smaller perturbing bend in TM7, was indeed found to allow partial rescue of transport activity. The double mutation T381V-V382P, on the other hand, did not rescue transport activity. Computational analysis of this mutant predicted a markedly different conformational preference from either the V382P or the T381A-V382P mutants. These studies show that changes in the structure of TM7 exert a strong influence on SERT's ability to achieve a mature, properly folded, cell surface conformation. PMID- 12779334 TI - Copper coordination in the full-length, recombinant prion protein. AB - The prion protein (PrP) binds divalent copper at physiologically relevant conditions and is believed to participate in copper regulation or act as a copper dependent enzyme. Ongoing studies aim at determining the molecular features of the copper binding sites. The emerging consensus is that most copper binds in the octarepeat domain, which is composed of four or more copies of the fundamental sequence PHGGGWGQ. Previous work from our laboratory using PrP-derived peptides, in conjunction with EPR and X-ray crystallography, demonstrated that the HGGGW segment constitutes the fundamental binding unit in the octarepeat domain [Burns et al. (2002) Biochemistry 41, 3991-4001; Aronoff-Spencer et al. (2000) Biochemistry 39, 13760-13771]. Copper coordination arises from the His imidazole and sequential deprotonated glycine amides. In this present work, recombinant, full-length Syrian hamster PrP is investigated using EPR methodologies. Four copper ions are taken up in the octarepeat domain, which supports previous findings. However, quantification studies reveal a fifth binding site in the flexible region between the octarepeats and the PrP globular C-terminal domain. A series of PrP peptide constructs show that this site involves His96 in the PrP(92 96) segment GGGTH. Further examination by X-band EPR, S-band EPR, and electron spin-echo envelope spectroscopy, demonstrates coordination by the His96 imidazole and the glycine preceding the threonine. The copper affinity for this type of binding site is highly pH dependent, and EPR studies here show that recombinant PrP loses its affinity for copper below pH 6.0. These studies seem to provide a complete profile of the copper binding sites in PrP and support the hypothesis that PrP function is related to its ability to bind copper in a pH-dependent fashion. PMID- 12779335 TI - Preparation and characterization of a 5'-deazaFAD T491V NADPH-cytochrome P450 reductase. AB - NADPH-cytochrome P450 reductase is a flavoprotein which contains both an FAD and FMN cofactor. Since the distribution of electrons is governed solely by the redox potentials of the cofactors, there are nine different ways the electrons can be distributed and hence nine possible unique forms of the protein. More than one species of reductase will exist at a given level of oxidation except when the protein is either totally reduced or oxidized. In an attempt to unambiguously characterize the redox properties of the physiologically relevant FMNH(2) form of the reductase, the T491V mutant of NADPH-cytochrome P450 reductase has been reconstituted with 5'-deazaFAD which binds to the FAD-binding site of the reductase with a K(d) of 94 nM. The 5'-deazaFAD cofactor does not undergo oxidation or reduction under our experimental conditions. The molar ratio of FMN to 5'-deazaFAD in the reconstituted reductase was 1.1. Residual FAD accounted for less than 5% of the total flavins. Addition of 2 electron equivalents to the 5' deazaFAD T491V reductase from dithionite generated a stoichiometric amount of the FMN hydroquinone form of the protein. The 5'-deazaFAD moiety remained oxidized under these conditions due to its low redox potential (-650 mV). The 2-electron reduced 5'-deazaFAD reductase was capable of transferring only a single electron from its FMN domain to its redox partners, ferric cytochrome c and cytochrome b(5). Reduction of the cytochromes and oxidation of the reductase occurred simultaneously. The FMNH(2) in the 5'-deazaFAD reductase autoxidizes with a first order rate constant of 0.007 s(-)(1). Availability of a stable NADPH-cytochrome P450 reductase capable of donating only a single electron to its redox partners provides a unique tool for investigating the electron-transfer properties of an intact reductase molecule. PMID- 12779337 TI - Equilibrium folding of the core histones: the H3-H4 tetramer is less stable than the H2A-H2B dimer. AB - To compare the stability of structurally related dimers and to aid in understanding the thermodynamics of nucleosome assembly, the equilibrium stabilities of the recombinant wild-type H3-H4 tetramer and H2A-H2B dimer have been determined by guanidinium-induced denaturation, using fluorescence and circular dichroism spectroscopies. The unfolding of the tetramer and dimer are highly reversible. The unfolding of the H2A-H2B dimer is a two-state process, with no detected equilibrium intermediates. The H3-H4 tetramer is unstable at moderate ionic strengths (mu approximately 0.2 M). TMAO (trimethylamine-N-oxide) was used to stabilize the tetramer; the stability of the H2A-H2B dimer was determined under the same solvent conditions. The equilibrium unfolding of H3-H4 was best described by a three-state mechanism, with well-folded H3-H4 dimers as a populated intermediate. When compared to H2A-H2B, the H3-H3 tetramer interface and the H3-H4 histone fold are strikingly less stable. The free energy of unfolding, in the absence of denaturant, for the H3-H4 and H2A-H2B dimers are 12.4 and 21.0 kcal mol(-)(1), respectively, in 1 M TMAO. It is postulated that the difference in stability between the histone dimers, which contain the same fold, is the result of unfavorable tertiary interactions, most likely the partial to complete burial of three salt bridges and burial of a charged hydrogen bond. Given the conservation of these buried interactions in histones from yeast to mammals, it is speculated that the H3-H4 tetramer has evolved to be unstable, and this instability may relate to its role in nucleosome dynamics. PMID- 12779336 TI - Modulation of the active complex assembly and turnover rate by protein-DNA interactions in Cre-LoxP recombination. AB - Cre promotes recombination at the 34 bp LoxP sequence. Substitution of a critical C-G base pair in LoxP with an A-T base pair, to give LoxAT, reduced Cre binding in vitro and abolished recombination in vivo [Hartung, M., and Kisters-Woike, B. (1998) J. Biol. Chem. 273, 22884-22891].We demonstrated that LoxAT can be recombined in vitro. However, Cre discriminates against this substrate both before and after DNA binding. The preference for LoxP over LoxAT is the result of reduced binding and a slower turnover rate, amplified by changes in cooperativity of complex assembly. With LoxAT, similar levels of substrate turnover required 2 2.5-fold higher protein-DNA concentrations compared to LoxP, but the sigmoidal behavior of the concentration dependence was more pronounced. Further, the Cre LoxAT complexes reacted 4-5-fold more slowly. In the 2.3 A resolution Cre-LoxAT complex structure, the major groove Arg259-guanine interaction was disrupted, explaining the reduced binding. Overall structural shifts and mobility changes indicate more favorable interactions between subunits, providing a hypothesis for the reduced turnover rate. Concomitant with the displacement of Arg259 from the DNA, adjacent charged residues Glu262 and Glu266 shifted to form salt bridges with the Arg259 guanidinium moiety. Substitution of Glu262 and Glu266 with glutamine increased Cre complex assembly efficiency and reaction rates with both LoxAT and LoxP, but diminished Cre's ability to distinguish them. The increased rate of this variant suggests that DNA substrate binding and turnover are coupled. The improved efficiency, made at some expense of sequence discrimination, may be useful for enhancing recombination in vivo. PMID- 12779338 TI - Experimental investigation of initial steps of helix propagation in model peptides. AB - It is not certain whether the helix propagation parameters s(n)() (i.e., the equilibrium constants between (n - 1)- and n-residue long alpha-helices) determined from numerous studies of rather long model peptides are applicable for description of the initial steps of the helix formation during the protein folding process. From fluorescence, NMR, and calorimetric studies of a series of model peptides, containing the La(3+)-binding sequence nucleating the helix (Siedlecka, M., Goch, G., Ejchart, A., Sticht, H., and Bierzynski, A. (1999) Proc. Natl. Acad. Sci. U.S.A. 96, 903-908), we have determined, at 25 degrees C, the average values of the enthalpy DeltaH(n)() and of the helix growth parameters s(n)() describing the first four steps of helix propagation in polyalanine. The absolute values of the C-cap parameters, describing the contribution of the C terminal residues to the helix free energy, have also been estimated for alanine (1.2 +/- 0.5) and NH(2) group (1.6 +/- 0.7). The initial four steps of the helix growth in polyalanine can be described by a common propagation parameter s = 1.54 +/- 0.04. The enthalpy DeltaH(n)() is also constant and equals -980 +/- 100 cal mol(-)(1). PMID- 12779339 TI - A familiar motif in a new context: the catalytic mechanism of hydroxyisourate hydrolase. AB - Hydroxyisourate hydrolase is a recently discovered enzyme that participates in the ureide pathway in soybeans. Its role is to catalyze the hydrolysis of 5 hydroxyisourate, the product of the urate oxidase reaction. There is extensive sequence homology between hydroxyisourate hydrolase and retaining glycosidases; in particular, the conserved active site glutamate residues found in retaining glycosidases are present in hydroxyisourate hydrolase as Glu 199 and Glu 408. However, experimental investigation of their roles, as well as the catalytic mechanism of the enzyme, have been precluded by the instability of 5 hydroxyisourate. Here, we report that diaminouracil serves as a slow, alternative substrate and can be used to investigate catalysis by hydroxyisourate hydrolase. The activity of the E199A protein was reduced 400-fold relative to wild-type, and no activity could be detected with the E408A mutant. Steady-state kinetic studies of the wild-type protein revealed that the pH-dependence of V(max) and V/K describe bell-shaped curves, consistent with the hypothesis that catalysis requires two ionizable groups in opposite protonation states. Addition of 100 mM azide accelerated the reaction catalyzed by the wild-type enzyme 8-fold and the E199A mutant 20-fold but had no effect on the E408A mutant. These data suggest that Glu 408 acts as a nucleophile toward the substrate forming a covalent anhydride intermediate, and Glu 199 facilitates formation of the intermediate by serving as a general acid and then activates water for hydrolysis of the intermediate. Thus, the mechanism of hydroxyisourate hydrolase is strikingly similar to that of retaining glycosidases, even though it catalyzes hydrolysis of an amide bond. PMID- 12779340 TI - Introduction of a pi-pi interaction at the active site of a cupredoxin: characterization of the Met16Phe Pseudoazurin mutant. AB - The Met16Phe mutant of the type 1 copper protein pseudoazurin (PACu), in which a phenyl ring is introduced close to the imidazole moiety of the His81 ligand, has been characterized. NMR studies indicate that the introduced phenyl ring is parallel to the imidazole group of His81. The mutation has a subtle effect on the position of the two S(Cys)-->Cu(II) ligand-to-metal charge transfer bands in the visible spectrum of PACu(II) and a more significant influence on their intensities resulting in a A(459)/A(598) ratio of 0.31 for Met16Phe as compared to a A(453)/A(594) ratio of 0.43 for wild-type PACu(II) at pH 8. The electron paramagnetic resonance spectrum of the Met16Phe variant is more axial than that of the wild-type protein, and the resonance Raman spectrum of the mutant exhibits subtle differences. A C(gamma)H proton of Met86 exhibits a much smaller hyperfine shift in the paramagnetic (1)H NMR spectrum of Met16Phe PACu(II) as compared to its position in the wild-type protein, which indicates a weaker axial Cu-S(Met86) interaction in the mutant. The Met16Phe mutation results in an approximately 60 mV increase in the reduction potential of PACu. The pK(a) value of the ligand His81 decreases from 4.9 in wild-type PACu(I) to 4.5 in Met16Phe PACu(I) indicating that the pi-pi contact with Phe16 stabilizes the Cu-N(His81) interaction. The Met16Phe variant of PACu has a self-exchange rate constant at pH 7.6 (25 degrees C) of 9.8 x 10(3) M(-)(1) s(-)(1) as compared to the considerably smaller value of 3.7 x 10(3) M(-)(1) s(-)(1) for the wild-type protein under identical conditions. The enhanced electron transfer reactivity of Met16Phe PACu is a consequence of a lower reorganization energy due to additional active site rigidity caused by the pi-pi interaction between His81 and the introduced phenyl ring. PMID- 12779341 TI - Mechanism of posttranslational regulation of phenol sulfotransferase: expression of two enzyme forms through redox modification and nucleotide binding. AB - Sulfotransferase catalyzes sulfuryl group transfer between a nucleotide and a variety of nucleophiles that may be sugar, protein, xenobiotics, and other small molecules. Nucleotides may serve as cosubstrate, cofactor, inhibitor, or regulator in an enzyme catalyzed sulfuryl group transfer reaction. We are trying to understand how nucleotide regulates the activity of phenol sulfotransferase (PST) through the expression of two enzyme forms. The homogeneous rat recombinant PST was obtained from Escherichia coli, and the nucleotide copurified was examined. The nucleotide was completely removed from inactive PST in high salt and oxidative condition. Total enzyme activity was recovered following incubation in reductive environment. Many nucleotides are known to tightly bind to PST but only one nucleotide, 3'-phosphoadenosine 5'-phosphate (PAP), was identified to combine with PST by ion-pair RP-HPLC, UV-visible spectra, (31)P NMR, and ESI-MS and MS-MS spectrometry. In addition to the presence or absence of PAP, oxidation following reduction of PST was required to completely interconvert the two forms of PST. According to the experimental results, a mechanism for the formation of the two enzyme forms was proposed. PMID- 12779343 TI - Structural studies of N- and C-terminally truncated human apolipoprotein A-I. AB - Apolipoprotein A-I (apoA-I) plays an important structural and functional role in lipid transport and metabolism. This work is focused on the central region of apoA-I (residues 60-183) that is predicted to contain exclusively amphipathic alpha-helices. Six N- and/or C-terminally truncated mutants, delta(1-41), delta(1 59), delta(198-243), delta(209-243), delta(1-41,185-243), and delta(1-59,185 243), were analyzed in their lipid-free state in solution at pH 4.7-7.8 by far- and near-UV CD spectroscopy. At pH 7.8, all mutants show well-defined secondary structures consisting of 40-52% alpha-helix. Comparison of the alpha-helix content in the wild type and mutants suggests that deletion of either the N- or C terminal region induces helical unfolding elsewhere in the structure, indicating that the terminal regions are important for the integrity of the solution conformation of apoA-I. Near-UV CD spectra indicate significant tertiary and/or quaternary structural changes resulting from deletion of the N-terminal 41 residues. Reduction in pH from 7.8 to 4.7 leads to an increase in the mutant helical content by 5-20% and to a large increase in thermal unfolding cooperativity. Van't Hoff analysis of the mutants at pH 4.7 indicates melting temperatures T(m) ranging from 51 to 59 degrees C and effective enthalpies deltaH(v)(T(m)) = 35 +/- 5 kcal/mol, similar to the values for plasma apoA-I at pH 7.8 (T(m) = 57 degrees C, deltaH(v) = 32 kcal/mol). Our results provide the first report of the pH effects on the secondary, tertiary, and/or quaternary structure of apoA-I variants and indicate the importance of the electrostatic interactions for the solution conformation of apoA-I. PMID- 12779342 TI - Role of radical formation at tyrosine 193 in the allene oxide synthase domain of a lipoxygenase-AOS fusion protein from coral. AB - Coral allene oxide synthase (cAOS), a fusion protein with 8R-lipoxygenase in Plexaura homomalla, is a hemoprotein with sequence similarity to catalases. cAOS reacts rapidly with the oxidant peracetic acid to form heme compound I and intermediate II. Concomitantly, an electron paramagnetic resonance (EPR) signal with tyrosyl radical-like features, centered at a g-value of 2.004-2.005, is formed. The radical is identified as tyrosyl by changes in EPR spectra when deuterated tyrosine is incorporated in cAOS. The radical location in cAOS is determined by mutagenesis of Y193 and Y209. Upon oxidation, native cAOS and mutant Y209F exhibit the same radical spectrum, but no significant tyrosine radical forms in mutant Y193H, implicating Y193 as the radical site in native cAOS. Estimates of the side chain torsion angles for the radical at Y193, based on the beta-proton isotropic EPR hyperfine splitting, A(iso), are theta(1) = 21 to 30 degrees and theta(2) = -99 to -90 degrees. The results show that cAOS can cleave nonsubstrate hydroperoxides by a heterolytic path, although a homolytic course is likely taken in converting the normal substrate, 8R hydroperoxyeicosatetraenoic acid (8R-HpETE), to product. Coral AOS achieves specificity for the allene oxide formed by selection of the homolytic pathway normally, while it inactivates by the heterolytic path with nonoptimal substrates. Accordingly, with the nonoptimal substrate, 13R hydroperoxyoctadecadienoic acid (13R-HpODE), mutant Y193H is inactivated after turning over significantly fewer substrate molecules than required to inactivate native cAOS or the Y209F mutant because it cannot absorb oxidizing equivalents by forming a radical at Y193. PMID- 12779344 TI - Combinatorial determination of sequence specificity for nanomolar DNA-binding hairpin polyamides. AB - Development of sequence-specific DNA-binding drugs is an important pharmacological goal, given the fact that numerous existing DNA-directed chemotherapeutic drugs rely on the strength and selectivity of their DNA interactions for therapeutic activity. Among the DNA-binding antibiotics, hairpin polyamides represent the only class of small molecules that can practically bind any predetermined DNA sequence. DNA recognition by these ligands depends on their side-by-side amino acid pairings in the DNA minor groove. Extensive studies have revealed that these molecules show extremely high affinity for sequence-directed, minor groove interaction. However, the specificity of such interactions in the presence of a large selection of sequences such as the human genome is not known. We used the combinatorial selection method restriction endonuclease protection, selection, and amplification (REPSA) to determine the DNA binding specificity of two hairpin polyamides, ImPyPyPy-gamma-PyPyPyPy-beta-Dp and ImPyPyPy-gamma ImPyPyPy-beta-Dp, in the presence of more than 134 million different sequences. These were verified by restriction endonuclease protection assays and DNase I footprinting analysis. Our data showed that both hairpin polyamides preferentially selected DNA sequences having consensus recognition sites as defined by the Dervan pairing rules. These consensus sequences were rather degenerate, as expected, given that the stacked pyrrole-pyrrole amino acid pairs present in both polyamides are unable to discriminate between A.T and T.A base pairs. However, no individual sequence within these degenerate consensus sequences was preferentially selected by REPSA, indicating that these hairpin polyamides are truly consensus-specific DNA-binding ligands. We also discovered a preference for overlapping consensus binding sites among the sequences selected by the hairpin polyamide ImPyPyPy-gamma-PyPyPyPy-beta-Dp, and confirmed by DNase I footprinting that these complex sites provide higher binding affinity. These data suggest that multiple hairpin polyamides can cooperatively bind to their highest-affinity sites. PMID- 12779345 TI - A novel alpha-conotoxin identified by gene sequencing is active in suppressing the vascular response to selective stimulation of sensory nerves in vivo. AB - We describe the identification of a conopeptide sequence in venom duct mRNA from Conus victoriae that suppresses a vascular response to pain in the rat. PCR-RACE was used to screen venom duct cDNAs for those transcripts that encode specific antagonists of vertebrate neuronal nicotinic acetylcholine receptors (nAChRs). One of these peptides, Vc1.1, was active as an antagonist of neuronal nAChRs in receptor binding and functional studies in bovine chromaffin cells. It also suppressed the vascular responses to unmyelinated sensory nerve C-fiber activation in rats. Such vascular responses are involved in pain transmission. Furthermore, its ability to suppress C-fiber function was greater than that of MVIIA, an omega-conotoxin with known analgesic activity in rats and humans. Vc1.1 has a high degree of sequence similarity to the alpha-conotoxin family of peptides and has the 4,7 loop structure characteristic of the subfamily of peptides that act on neuronal-type nAChRs. The results suggest that neuronal alpha-conotoxins should be further investigated with respect to their potential to suppress pain. PMID- 12779346 TI - The ATPase activity of the ChlI subunit of magnesium chelatase and formation of a heptameric AAA+ ring. AB - The AAA(+) ATPase component of magnesium chelatase (ChlI) drives the insertion of Mg(2+) into protoporphyrin IX; this is the first step in chlorophyll biosynthesis. We describe the ATPase activity, nucleotide binding kinetics, and structural organization of the ChlI protein. A consistent reaction scheme arises from our detailed steady state description of the ATPase activity of the ChlI subunit and from transient kinetic analysis of nucleotide binding. We provide the first demonstration of metal ion binding to a specific subunit of any of the multimeric chelatases and characterize binding of Mg(2+) to the free and MgATP(2)(-) bound forms of ChlI. Transient kinetic studies with the fluorescent substrate analogue TNP-ATP show that there are two forms of monomeric enzyme, which have distinct magnesium binding properties. Additionally, we describe the self-association properties of the subunit and provide a structural analysis of the multimeric ring formed by this enzyme in the presence of nucleotide. This single particle analysis demonstrates that this species has a 7-fold rotational symmetry, which is in marked contrast to most members of the AAA(+) family that tend to form hexamers. PMID- 12779347 TI - Direct binding of the N-terminus of HTLV-1 tax oncoprotein to cyclin-dependent kinase 4 is a dominant path to stimulate the kinase activity. AB - The involvement of Tax oncoprotein in the INK4-CDK4/6-Rb pathway has been regarded as a key factor for immortalization and transformation of human T-cell leukemia virus 1 (HTLV-1) infected cells. In both p16 -/- and +/+ cells, expression of Tax has been correlated with an increase in CDK4 activity, which subsequently increases the phosphorylation of Rb and drives the infected cells into cell cycle progression. In relation to these effects, Tax has been shown to interact with two components of the INK4-CDK4/6-Rb pathway, p16 and cyclin D(s). While Tax competes with CDK4 for p16 binding, thus suppressing p16 inhibition of CDK4, Tax also binds to cyclin D(s) with concomitant increases in both CDK4 activity and the phosphorylation of cyclin D(s). Here we show that both Tax and residues 1-40 of the N-terminus of Tax, Tax40N, bind to and activate CDK4 in vitro. In the presence of INK4 proteins, binding of Tax and Tax40N to CDK4 counteracts against the inhibition of p16 and p18 and acts as the major path to regulate Tax-mediated activation of CDK4. We also report that Tax40N retains the transactivation ability. These results of in vitro studies demonstrate a potentially novel, p16-independent route to regulate CDK4 activity by the Tax oncoprotein in HTLV-1 infected cells. PMID- 12779349 TI - Cytokines and antitumor immunity. AB - Currently, the notion of immunosurveillance against tumors is enjoying something of a renaissance. Even if we still refuse to accept that tumors arising in the normal host are unable to trigger an immune response because of the lack of initiation ("danger") signals, there is no doubt that the immune system can be manipulated experimentally and by implication therapeutically to exert anti-tumor effects. For this activity to be successful, the appropriate cytokine milieu has to be provided, making cytokine manipulation central to immunotherapy. On the other hand, the major hurdle currently preventing successful immunotherapy is the ability of tumors to evolve resistant variants under the pressure of immune selection. Here, too, the cytokine milieu plays an essential role. The purpose of this brief review is to consider the current status of the application of cytokines in facilitating antitumor immunity, as well their role in inhibiting responses to tumors. Clearly, encouraging the former but preventing the latter will be the key to the effective clinical application of cancer immunotherapy. PMID- 12779350 TI - Solid tumor therapy: manipulation of the vasculature with TNF. AB - Drug delivery to solid tumors is one of the most challenging aspects in cancer therapy. Whereas agents seem promising in the test tube, clinical trials often fail due to unfavorable pharmacokinetics, poor delivery, low local concentrations, and limited accumulation in the target cell. A major step forwards in the treatment of solid tumors is the recognition of the tumor associated vasculature as an important target for therapy. Inhibition of tumor vascular development has a direct effect on the growth and progression of the tumor. Destruction of an existing vasculature also directly inflicts serious damage to the tumor cell. Moreover, the tumor vascular bed can be manipulated facilitating enhanced permissiveness of the tumor for administered chemotherapeutics. In this review, we focus on the use of tumor necrosis factor alpha (TNF) in local and systemic therapy in conjunction with chemotherapy. In these settings TNF demonstrates potent and selective activity on the tumor vascular bed, which strongly improves tumor response. PMID- 12779351 TI - The timing of immunotherapy and nephrectomy in multimodality treatment of metastatic renal cell carcinoma. AB - Multimodality treatment of metastatic renal cell carcinoma with immunotherapy and cytoreductive surgery is controversial. Especially the benefit of removing asymptomatic primary tumors in synchronous metastatic renal cell cancer has been debated since several non-randomized, retrospective studies revealed an improved response to immunotherapy and prolonged survival following initial nephrectomy. Two recent randomized prospective trials both demonstrated a prolonged survival in those who were randomly assigned to undergo nephrectomy of the primary tumor prior to treatment with interferon alfa-2b than in those who were assigned to undergo treatment with interferon alfa-2b alone. In these trials the survival benefit was limited and strongly influenced by overall performance score. The timing of immunotherapy, either as neoadjuvant (prior to nephrectomy) or adjuvant treatment (following nephrectomy) in the multimodality approach of synchronous metastatic renal cell carcinoma remains controversial. Selection of patients, the possible mechanisms underlying the survival advantage of the combination of nephrectomy and immunotherapy, and the timing of the treatment modalities are discussed herein. PMID- 12779352 TI - IL-2 gene and antisense TGF-beta1 strategies counteract HSV-2 transformed tumor progression. AB - The H238 tumor cells are Herpes simplex virus type 2-transformed BALB/c mouse fibroblasts that constitutively express transforming growth factor (TGF-beta1). TGF-beta can diminish immune capacity, whereas interleukin 2 (IL-2) is stimulatory to the immune system and can counteract the negative effects of TGF beta1. The H238-BALB/c system provides a syngeneic model to evaluate new strategies with the potential to ameliorate tumor-induced immune depression. Plasmids expressing either antisense TGF-beta1 or murine Il-2 were constructed and stably transformed cells generated (masH238 and H238-IL2, respectively). In vitro measurements (ELISA and RT-PCR) demonstrated a >70% decrease in TGF-beta1 secretion by the masH238 tumor cells, and significant levels of IL-2 production by the H238-IL2 transfected cells when compared to wild type and control plasmid transfected H238 cells. BALB/c mice injected subcutaneously with the masH238 cells developed significantly smaller tumors than the controls. Mice injected with H238-IL-2 cells developed tumors that failed to progress relative to control tumor growth. The differences in tumor growth in the mice were associated with enhanced immune reactivity and an increased response to T lymphocyte mitogens. Significant differences were also noted in lymphocyte populations and expression of CD25 and CD71 activation markers in the blood and spleens of mice receiving transfected tumor cells. Collectively, the data demonstrate that strategies employing antisense TGF-beta1 and IL-2 expression by transfected tumor cells can counteract the progression of a TGF-beta1-secreting tumor and enhance immune function involving modulation of T lymphocyte populations. PMID- 12779353 TI - Radiation enhances the anti-tumor effects of vaccinia-p53 gene therapy in glioma. AB - The overall goal of this study was to analyze the effect and mechanism of radiation in combination with vaccinia viruses (VV) carrying the p53 gene against glioma. Comparison of two alternative treatments of cultured C6 (p53(+)) and 9L (p53(-)) rat glioma cells showed significantly reduced survival for both cell lines, especially 9L, when radiation was applied prior to virus versus radiation alone. High p53 protein expression mediated by VV-TK-p53 was measured in infected cells. Single modality treatment of C6 cells with psoralen and UV (PUV) inactivated VV-TK-p53 (PUV-VV-TK-53) or radiation significantly decreased survival compared with PUV-inactivated L-15 (PUV-L-15) control virus. However, no difference was observed between radiation and combination treatments of C6 cells. In contrast, radiation followed by PUV-VV-TK-53 resulted in dramatic reduction of 9L cell viability, compared to single modality treatment. Flow cytometry analysis of Annexin-V-stained 9L cells showed that radiation and PUV-VV-TK-53 caused a significant decrease in live cells (17.2%) as compared to other treatments and control (61.6-98.3%). Apoptosis was observed in 37.2% of cells, while the range was 0.7-7.8% in other treatment groups; maximal p53 level was measured on day 7 post-infection. In athymic mice bearing C6 tumors, VV-TK-53 plus radiation in both single and multiple therapies resulted in significantly smaller tumors by day 30 compared to the agents given only once. Immunohistochemical analysis of tumor sections demonstrated p53 protein expression over 20 days after VV-TK-53 treatment. Analysis of blood and spleen cells of mice given multiple combination treatments showed significant splenomegaly, leukocytosis, and increased DNA synthesis and response to mitogen. Multiple combination treatments were also associated with significantly elevated natural killer and B cells in the spleen. There were no overt toxicities, although depression in red blood cell and thrombocyte parameters was noted. Collectively, the data demonstrate that radiation significantly improves the efficacy of VV-mediated tumor suppressor p53 therapy and may be a promising strategy for glioma treatment. Furthermore, the results support the conclusion that the mechanisms underlying the enhanced anti tumor effect of combination treatment include apoptosis/necrosis and upregulation of innate immune defenses. PMID- 12779354 TI - Immunotherapy of cancer by active vaccination: does allogeneic bone marrow transplantation after non-myeloablative conditioning provide a new option? AB - The critical role of antigen-specific T cells in cancer immunotherapy has been amply demonstrated in many model systems. Though success of clinical trials still remains far behind expectation, the continuous improvement in our understanding of the biology of the immune response will provide the basis of optimized cancer vaccines and allow for new modalities of cancer treatment. This review focuses on the current status of active therapeutic vaccination and future prospects. The latter will mainly be concerned with allogeneic bone marrow cell transplantation after non-myeloablative conditioning, because it is my belief that this approach could provide a major breakthrough in cancer immunotherapy. Concerning active vaccination protocols the following aspects will be addressed: i) the targets of immunotherapeutic approaches; ii) the response elements needed for raising a therapeutically successful immune reaction; iii) ways to achieve an optimal confrontation of the immune system with the tumor and iv) supportive regimen of immunomodulation. Hazards which one is most frequently confronted with in trials to attack tumors with the inherent weapon of immune defense will only be briefly mentioned. Many question remain to be answered in the field of allogeneic bone marrow transplantation after non-myeloablative conditioning to optimize the therapeutic setting for this likely very powerful tool of cancer therapy. Current considerations to improve engraftment and to reduce graft versus host disease while strengthening graft versus tumor reactivity will be briefly reviewed. Finally, I will discuss whether tumor-reactive T cells can be "naturally" maintained during the process of T cell maturation in the allogeneic host. Provided this hypothesis can be substantiated, a T cell vaccine will meet a pool of virgin T cells in the allogeneically reconstituted host, which are tolerant towards the host, but not anergised towards tumor antigens presented by MHC molecules of the host. PMID- 12779355 TI - The efficacy of linear accelerator stereotactic radiosurgery in treating glomus jugulare tumors. AB - Treatment of glomus jugulare tumors with radiosurgery has grown in acceptance since the first reported treatment in 1995, but only a few centers have reported their experiences with limited follow up time. We report our experience with stereotactic radiosurgery in nine patients with ten glomus tumors. All patients were treated either with frame based LINAC or Cyberknife with doses ranging from 1600 cGy to 2500 cGy. Three patients received no previous therapy and one patient received additional external beam radiation for concomitant treatment of carotid body tumors. Patients were then followed for post treatment side effects in addition to change in tumor size by MRI evaluation. The median clinical follow-up time was 26 months (mean 54 months), median radiographic follow-up was 21.5 months (mean 46 months), with a range from 3 to 126 months. The results from our center demonstrated nine of ten tumors to be stable in size by MRI exam, and one tumor which regressed in size. Nine patients had stable neurological symptoms, and one patient experienced transient ipsilateral tongue weakness and hearing loss, both of which subsequently resolved. Our results continue to support radiosurgery as a suitable form of treatment for glomus jugulare tumors as evidenced by results from this four and a half year follow-up. PMID- 12779356 TI - Factors associated with improved survival in patients with brain metastases from esophageal cancer: a retrospective review. AB - There are over 200,000 cases of brain metastases (BrM) every year, but very few are from esophageal cancer primaries. In order to determine predictors for outcome of these patients, the authors conducted a retrospective review of twenty seven patients with BrM from esophageal carcinoma diagnosed at the Cleveland Clinic Foundation between 1991 and 2001. For the entire cohort, median follow-up and median survival was 3.6 months and 3.6 months, respectively. On univariate analysis, patients with Karnofsky Performance Status (KPS) >/= 70, low recursive partitioning analysis score, single BrM, no systemic disease, and aggressive treatment [surgery, stereotactic radiosurgery (SRS) + whole brain radiation (WBRT), SRS + surgery + WBRT, surgery + WBRT)] had a significantly improved survival. In a multivariate model, patients with higher KPS and aggressive treatment had improved survival. The 1-year survival for the WBRT alone group and the aggressive treatment group was 6%, and 36% respectively. We conclude that based on the data presented here, patients with BrM from esophageal cancer have poor outcome. Aggressive treatment and favorable KPS are associated with longer survival for selected patients. We recommend esophageal cancer patients with BrM be enrolled in clinical trials to better delineate the role of treatment and potentially improve results. PMID- 12779357 TI - Virtual bronchoscopy: comparison of different surface rendering models. AB - The aim of this study was to compare different representation models of surface rendered virtual bronchoscopy. 10 consecutive patients with inoperable primary lung tumors underwent thin-section spiral computed tomography. The structures of interest, the tracheobronchial system and anatomical and pathological thoracic structures were segmented using an interactive threshold interval volume-growing segmentation algorithm and visualized with the aid of a color-coded surface rendering method. For virtual bronchoscopy, the tracheobronchial system was visualized using a triangle-surface rendering model, a shaded-surface rendering model and a transparent shaded-surface rendering model. The triangle-surface rendering model allowed optimum detailed spatial representation of the dimensions of extraluminal anatomical and pathological mediastinal structures. As the lumen of the tracheobronchial system was less well defined, the rendering model was of limited use for depiction of the airway surface. The shaded-surface rendering model facilitated an optimum assessment of the airway surface, but the mediastinal structures could not be depicted. The transparent shaded-surface rendering model provides simultaneous adequate to optimum visualization and assessment of the intraluminal airway surface and the extraluminal mediastinal structures as well as a quantitative assessment of the spatial relationship between these structures. Fast data acquisition with a multi-slice detector spiral computed tomography scanner and the use of virtual bronchoscopy with the transparent shaded-surface rendering model obviate the need for time consuming detailed analysis and presentation of axial source images by providing improved the diagnostic imaging of endotracheal and endobronchial diseases and offering a useful alternative to fiberoptic bronchoscopy. PMID- 12779358 TI - Chaotic kinetics and transport (Overview). PMID- 12779359 TI - Nonlinear instability of elementary stratified flows at large Richardson number. AB - Elementary stably stratified flows with linear instability at all large Richardson numbers have been introduced recently by the authors [J. Fluid Mech. 376, 319-350 (1998)]. These elementary stratified flows have spatially constant but time varying gradients for velocity and density. Here the nonlinear stability of such flows in two space dimensions is studied through a combination of numerical simulations and theory. The elementary flows that are linearly unstable at large Richardson numbers are purely vortical flows; here it is established that from random initial data, linearized instability spontaneously generates local shears on buoyancy time scales near a specific angle of inclination that nonlinearly saturates into localized regions of strong mixing with density overturning resembling Kelvin-Helmholtz instability. It is also established here that the phase of these unstable waves does not satisfy the dispersion relation of linear gravity waves. The vortical flows are one family of stably stratified flows with uniform shear layers at the other extreme and elementary stably stratified flows with a mixture of vorticity and strain exhibiting behavior between these two extremes. The concept of effective shear is introduced for these general elementary flows; for each large Richardson number there is a critical effective shear with strong nonlinear instability, density overturning, and mixing for elementary flows with effective shear below this critical value. The analysis is facilitated by rewriting the equations for nonlinear perturbations in vorticity-stream form in a mean Lagrangian reference frame. (c) 2000 American Institute of Physics. PMID- 12779360 TI - Eulerian mean flow from an instability of convective plumes. AB - The dynamical origin of large-scale flows in systems driven by concentrated Archimedean forces is considered. A two-dimensional model of plumes, such as those observed in thermal convection at large Rayleigh and Prandtl numbers, is introduced. From this model, we deduce the onset of mean flow as an instability of a convective state consisting of parallel vertical flow supported by buoyancy forces. The form of the linear equation governing the instability is derived and two modes of instability are discussed, one of which leads to the onset of steady Eulerian mean flow in the system. We are thus able to link the origin of mean flow precisely to the profiles of the unperturbed plumes. The form of the nonlinear partial differential equation governing the Eulerian mean flow, including nonlinear effects, is derived in one special case. The extension to three dimensions is outlined. (c) 2000 American Institute of Physics. PMID- 12779361 TI - Power spectrum of passive scalars in two dimensional chaotic flows. AB - In this paper the power spectrum of passive scalars transported in two dimensional chaotic fluid flows is studied theoretically. Using a wave-packet method introduced by Antonsen et al., several model flows are investigated, and the fact that the power spectrum has the k(-1)-scaling predicted by Batchelor is confirmed. It is also observed that increased intermittency of the stretching tends to make the roll-off of the power spectrum at the high k end of the k(-1) scaling range more gradual. These results are discussed in light of recent experiments where a k(-1) scaling range was not observed. (c) 2000 American Institute of Physics. PMID- 12779362 TI - Nonasymptotic properties of transport and mixing. AB - We study relative dispersion of passive scalar in nonideal cases, i.e., in situations in which asymptotic techniques cannot be applied; typically when the characteristic length scale of the Eulerian velocity field is not much smaller than the domain size. Of course, in such a situation usual asymptotic quantities (the diffusion coefficients) do not give any relevant information about the transport mechanisms. On the other hand, we shall show that the Finite Size Lyapunov Exponent, originally introduced for the predictability problem, appears to be rather powerful in approaching the nonasymptotic transport properties. This technique is applied in a series of numerical experiments in simple flows with chaotic behaviors, in experimental data analysis of drifter and to study relative dispersion in fully developed turbulence. (c) 2000 American Institute of Physics. PMID- 12779363 TI - Lattice models of advection-diffusion. AB - We present a synthesis of theoretical results concerning the probability distribution of the concentration of a passive tracer subject to both diffusion and to advection by a spatially smooth time-dependent flow. The freely decaying case is contrasted with the equilibrium case. A computationally efficient model of advection-diffusion on a lattice is introduced, and used to test and probe the limits of the theoretical ideas. It is shown that the probability distribution for the freely decaying case has fat tails, which have slower than exponential decay. The additively forced case has a Gaussian core and exponential tails, in full conformance with prior theoretical expectations. An analysis of the magnitude and implications of temporal fluctuations of the conditional diffusion and dissipation is presented, showing the importance of these fluctuations in governing the shape of the tails. Some results concerning the probability distribution of dissipation, and concerning the spatial scaling properties of concentration fluctuation, are also presented. Though the lattice model is applied only to smooth flow in the present work, it is readily applicable to problems involving rough flow, and to chemically reacting tracers. (c) 2000 American Institute of Physics. PMID- 12779364 TI - Self-consistent chaotic transport in fluids and plasmas. AB - Self-consistent chaotic transport is the transport of a field F by a velocity field v according to an advection-diffusion equation in which there is a dynamical constrain between the two fields, i.e., O(F,v)=0 where O is an integral or differential operator, and the Lagrangian trajectories of fluid particles exhibit sensitive dependence on initial conditions. In this paper we study self consistent chaotic transport in two-dimensional incompressible shear flows. In this problem F is the vorticity zeta, the corresponding advection-diffusion equation is the vorticity equation, and the self-consistent constrain is the vorticity-velocity coupling z nabla xv=zeta. To study this problem we consider three self-consistent models of intermediate complexity between the simple but limited kinematic chaotic advection models and the approach based on the direct numerical simulation of the Navier-Stokes equation. The first two models, the vorticity defect model and the single wave model, are constructed by successive simplifications of the vorticity-velocity coupling. The third model is an area preserving self-consistent map obtained from a space-time discretization of the single wave model. From the dynamical systems perspective these models are useful because they provide relatively simple self-consistent Hamiltonians (streamfunctions) for the Lagrangian advection problem. Numerical simulations show that the models capture the basic phenomenology of shear flow instability, vortex formation and relaxation typically observed in direct numerical simulations of the Navier-Stokes equation. Self-consistent chaotic transport in electron plasmas in the context of kinetic theory is also discussed. In this case F is the electron distribution function in phase space, the corresponding advection equation is the Vlasov equation and the self-consistent constrain is the Poisson equation. This problem is closely related to the vorticity problem. In particular, the vorticity defect model is analogous to the Vlasov-Poisson model and the single wave model and the self-consistent map apply equally to both plasmas and fluids. Also, the single wave model is analogous to models used in the study of globally coupled oscillator systems. (c) 2000 American Institute of Physics. PMID- 12779365 TI - Chaotic advection, diffusion, and reactions in open flows. AB - We review and generalize recent results on advection of particles in open time periodic hydrodynamical flows. First, the problem of passive advection is considered, and its fractal and chaotic nature is pointed out. Next, we study the effect of weak molecular diffusion or randomness of the flow. Finally, we investigate the influence of passive advection on chemical or biological activity superimposed on open flows. The nondiffusive approach is shown to carry some features of a weak diffusion, due to the finiteness of the reaction range or reaction velocity. (c) 2000 American Institute of Physics. PMID- 12779366 TI - Finding finite-time invariant manifolds in two-dimensional velocity fields. AB - For two-dimensional velocity fields defined on finite time intervals, we derive an analytic condition that can be used to determine numerically the location of uniformly hyperbolic trajectories. The conditions of our main theorem will be satisfied for typical velocity fields in fluid dynamics where the deformation rate of coherent structures is slower than individual particle speeds. We also propose and test a simple numerical algorithm that isolates uniformly finite-time hyperbolic sets in such velocity fields. Uniformly hyperbolic sets serve as the key building blocks of Lagrangian mixing geometry in applications. (c) 2000 American Institute of Physics. PMID- 12779367 TI - Heteroclinic primary intersections and codimension one Melnikov method for volume preserving maps. AB - We study families of volume preserving diffeomorphisms in R(3) that have a pair of hyperbolic fixed points with intersecting codimension one stable and unstable manifolds. Our goal is to elucidate the topology of the intersections and how it changes with the parameters of the system. We show that the "primary intersection" of the stable and unstable manifolds is generically a neat submanifold of a "fundamental domain." We compute the intersections perturbatively using a codimension one Melnikov function. Numerical experiments show various bifurcations in the homotopy class of the primary intersections. (c) 2000 American Institute of Physics. PMID- 12779368 TI - Transient chaotic mixing during a baroclinic life cycle. AB - We discuss how atmospheric eddies affect transport and mixing of tracers at midlatitudes. To this purpose, we study baroclinic life cycles in a simple dynamical model of the atmosphere. We consider the trapping properties of the developing eddies and the characteristics of meridional transport, and we identify regions of increased mixing. Although the flow is in principle three dimensional, we illustrate how some of the concepts developed in the study of two dimensional chaotic advection provide useful information on tracer dynamics in more complicated flows. (c) 2000 American Institute of Physics. PMID- 12779369 TI - Hierarchical structures in the phase space and fractional kinetics: I. Classical systems. AB - Hamiltonian chaotic dynamics is not ergodic due to the infinite number of islands imbedded in the stochastic sea. Stickiness of the islands' boundaries makes the wandering process very erratic with multifractal space-time structure. This complication of the chaotic process can be described on the basis of fractional kinetics. Anomalous properties of the chaotic transport become more transparent when there exists a set of islands with a hierarchical structure. Different consequences of the described phenomenon are discussed: a distribution of Poincare recurrences, characteristic exponents of transport, nonuniversality of transport, log periodicity, and chaos erasing. (c) 2000 American Institute of Physics. PMID- 12779370 TI - Hierarchical structures in the phase space and fractional kinetics: II. Immense delocalization in quantized systems. AB - Anomalous transport due to Levy-type flights in quantum kicked systems is studied. These systems are kicked rotor and kicked Harper model. It is confirmed for a kicked rotor that there exist special "magic" values of a control parameter of chaos K=K(*)=6.908 745 em leader for which an essential increasing of a localization length is obtained. Functional dependence of the localization length on both parameter of chaos and quasiclassical parameter h is studied. We also observe immense delocalization of the order of 10(9) for a kicked Harper model when a control parameter K is taken to be K(*)=6.349 972. This "magic" value corresponds to special phase space topology in the classical limit, when a hierarchical self-similar set of sticky islands emerges. The origin of the effect is of the general nature and similar immense delocalization as well as increasing of localization length can be found in other systems. (c) 2000 American Institute of Physics. PMID- 12779371 TI - Phase space structure and anomalous diffusion in a rotational fluid experiment. AB - The transport of passive scalars is considered in a model of rotating annulus experiments. The system has a chain of vortices and a jet, separated by a stochastic layer. For special values of the control parameters, the boundary of the stochastic layer can contain self-similar structures of islands with regular trajectories. Two such values are identified, with the structure being on the jet boundary and on the vortex boundary, respectively. The transport properties for both cases were studied by high-precision direct numerical integration of the equations of motion. The presence of such structures is found to significantly affect the statistical properties of the trajectories and the transport exponent. The results of the computations are compared with various theoretical models of anomalous diffusion. The particle behavior was found to depend significantly on the time scale, with different theories being applicable on different time intervals. Some regimes do not match any of the existing theories. (c) 2000 American Institute of Physics. PMID- 12779372 TI - Anomalous diffusion in two-dimensional potentials with hexagonal symmetry. AB - The diffusion process in a Hamiltonian dynamical system describing the motion of a particle in a two-dimensional (2D) potential with hexagonal symmetry is studied. It is shown that, depending on the energy of the particle, various transport processes can exist: normal (Brownian) diffusion, anomalous diffusion, and ballistic transport. The relationship between these transport processes and the underlying structure of the phase space of the Hamiltonian dynamical system is investigated. The anomalous transport is studied in detail in two particular cases: in the first case, inside the chaotic sea there exist self-similar structures with fractal properties while in the second case the transport takes place in the presence of multilayered structures. It is demonstrated that structures of the second type can lead to a physical situation in which the transport becomes ballistic. Also, it is shown that for all cases in which the diffusive transport is anomalous the trajectories of the diffusing particles contain long segments of regular motion, the length of these segments being described by Levy probability density functions. Finally, the numerical values of the parameters which describe the diffusion processes are compared with those predicted by existing theoretical models. (c) 2000 American Institute of Physics. PMID- 12779373 TI - Entropy computing via integration over fractal measures. AB - We discuss the properties of invariant measures corresponding to iterated function systems (IFSs) with place-dependent probabilities and compute their Renyi entropies, generalized dimensions, and multifractal spectra. It is shown that with certain dynamical systems, one can associate the corresponding IFSs in such a way that their generalized entropies are equal. This provides a new method of computing entropy for some classical and quantum dynamical systems. Numerical techniques are based on integration over the fractal measures. (c) 2000 American Institute of Physics. PMID- 12779374 TI - Spectral properties and anomalous transport in a polygonal billiard. AB - We analyze a class of polygonal billiards, whose behavior is conjectured to exhibit a variety of interesting dynamical features. Correlation functions are numerically investigated, and in a subclass of billiard tables they give indications about a singular continuous spectral measure. By lifting billiard dynamics we are also able to study transport properties: the (normal or anomalous) diffusive behavior is theoretically connected to a scaling index of the spectral measure; the proposed identity is shown to agree with numerical simulations. (c) 2000 American Institute of Physics. PMID- 12779375 TI - Statistical properties of actions of periodic orbits. AB - We investigate statistical properties of unstable periodic orbits, especially actions for two simple linear maps (p-adic Baker map and sawtooth map). The action of periodic orbits for both maps is written in terms of symbolic dynamics. As a result, the expression of action for both maps becomes a Hamiltonian of one dimensional spin systems with the exponential-type pair interaction. Numerical work is done for enumerating periodic orbits. It is shown that after symmetry reduction, the dyadic Baker map is close to generic systems, and the p-adic Baker map and sawtooth map with noninteger K are also close to generic systems. For the dyadic Baker map, the trace of the quantum time-evolution operator is semiclassically evaluated by employing the method of Phys. Rev. E 49, R963 (1994). Finally, using the result of this and with a mathematical tool, it is shown that, indeed, the actions of the periodic orbits for the dyadic Baker map with symmetry reduction obey the uniform distribution modulo 1 asymptotically as the period goes to infinity. (c) 2000 American Institute of Physics. PMID- 12779376 TI - Evaluation of the smoothed interference pattern under conditions of ray chaos. AB - A ray-based approach has been considered for evaluation of the coarse-grained Wigner function. From the viewpoint of wave propagation theory this function represents the local spectrum of the wave field smoothed over some spatial and angular scales. A very simple formula has been considered which expresses the smoothed Wigner function through parameters of ray trajectories. Although the formula is ray-based, it nevertheless has no singularities at caustics and its numerical implementation does not require looking for eigenrays. These advantages are especially important under conditions of ray chaos when fast growing numbers of eigenrays and caustics are the important factors spoiling applicability of standard semiclassical approaches already at short ranges. Similar factors restrict applicability of some semiclassical predictions in quantum mechanics at times exceeding the so-called "logarithm break time." Numerical calculations have been carried out for a particular model of range-dependent waveguide where ray trajectories exhibit chaotic motion. These calculations have confirmed our conjecture that by choosing large enough smoothing scales, i.e., by sacrificing small details of the interference pattern, one can substantially enhance the validity region of ray theory. (c) 2000 American Institute of Physics. PMID- 12779377 TI - Effect of interfacial tension on propagating polymerization fronts. AB - This paper is devoted to the investigation of polymerization fronts converting a liquid monomer into a liquid polymer. We assume that the monomer and the polymer are immiscible and study the influence of the interfacial tension on the front stability. The mathematical model consists of the reaction-diffusion equations coupled with the Navier-Stokes equations through the convection terms. The jump conditions at the interface take into account the interfacial tension. Simple physical arguments show that the same temperature distribution could not lead to Marangoni instability for a nonreacting system. We fulfill a linear stability analysis and show that interaction of the chemical reaction and of the interfacial tension can lead to an instability that has another mechanism: the heat produced by the reaction decreases the interfacial tension and initiates the liquid motion. It brings more monomer to the reaction zone and increases even more the heat production. This feedback mechanism can lead to the instability if the frontal Marangoni number exceeds a critical value. (c) 2000 American Institute of Physics. PMID- 12779378 TI - Homoclinic bifurcation in a Hodgkin-Huxley model of thermally sensitive neurons. AB - We study global bifurcations of the chaotic attractor in a modified Hodgkin Huxley model of thermally sensitive neurons. The control parameter for this model is the temperature. The chaotic behavior is realized over a wide range of temperatures and is visualized using interspike intervals. We observe an abrupt increase of the interspike intervals in a certain temperature region. We identify this as a homoclinic bifurcation of a saddle-focus fixed point which is embedded in the chaotic attractors. The transition is accompanied by intermittency, which obeys a universal scaling law for the average length of trajectory segments exhibiting only short interspike intervals with the distance from the onset of intermittency. We also present experimental results of interspike interval measurements taken from the crayfish caudal photoreceptor, which qualitatively demonstrate the same bifurcation structure. (c) 2000 American Institute of Physics. PMID- 12779379 TI - A method to Fourier filter textured images. AB - An algorithm is introduced to extract an underlying image from a class of textures. It is assumed that the image is bandwidth limited and the noise is broad-band. The initial step of the algorithm extends the signal to a larger periodic image using "Distributed Approximating Functionals." The second step introduces a low-pass filter which allows the identification and elimination of the high-frequency components of the noise. The periodicity of the resulting image allows it to be Fourier filtered without aliasing. The feasibility of the algorithm is demonstrated on several noisy patterns generated in experiments and model systems. (c) 2000 American Institute of Physics. PMID- 12779380 TI - Experiments on arrays of globally coupled chaotic electrochemical oscillators: Synchronization and clustering. AB - Experiments on chaotically oscillating arrays of 64 nickel electrodes in sulfuric acid were carried out. External resistors in parallel and series are added to vary the extent of global coupling among the oscillators without changing the other properties of the system. The array is heterogeneous due to small variations in the properties of the electrodes and there is also a small amount of noise. The addition of global coupling transforms a system of independent elements to a state of complete synchronization. At intermediate coupling strengths stable clusters, or condensates of elements, form. All the elements in a cluster follow the same chaotic trajectory but each cluster has its own dynamics; the system is thus temporally chaotic but spatially ordered. Many cluster configurations occur under the same conditions and transitions among them can be produced. For values of the coupling parameter on either side of the stable cluster region a non-stationary behavior occurs in which clustered and synchronized states alternately form and break up. Some statistical properties of the cluster states are determined. (c) 2000 American Institute of Physics. PMID- 12779381 TI - The correlation dimension of rat hearts in an experimentally controlled environment. AB - The electric response of several isolated rat hearts in a controlled environment was studied experimentally. The correlation dimension D(2) was estimated and was found to be between 4 and 6.5 when the response was nearly periodic. The variation of D(2) with the concentration of calcium was studied and a general trend of its increase with increasing concentration was found. Two types of ventricular fibrillation (VF) were observed, one that corresponds to a stochastic signal where D(2) is unbounded and the other to a low dimensional dynamical system with 3.5 infinity for three-dimensional flows and an increase of the gradient of the vorticity for planar and axisymmetric flows. Examples of such flows in the planar and axisymmetric cases are flows with a rectilinear streamline; this can be established using a special local Lyapunov function. Incompressible flows of a dusty medium are another example (it turns out that collapse is impossible for such flows, but the vorticity and the rate of deformation, as a rule, grow with no limit). Other examples can be constructed by composition of shear flows. It is shown that in the vorticity metric almost all stationary planar flows are unstable with respect to three-dimensional disturbances and in the vorticity gradient metric planar and axisymmetric flows with a rectilinear streamline are unstable. (c) 2000 American Institute of Physics. PMID- 12779421 TI - Resonantly forced inhomogeneous reaction-diffusion systems. AB - The dynamics of spatiotemporal patterns in oscillatory reaction-diffusion systems subject to periodic forcing with a spatially random forcing amplitude field are investigated. Quenched disorder is studied using the resonantly forced complex Ginzburg-Landau equation in the 3:1 resonance regime. Front roughening and spontaneous nucleation of target patterns are observed and characterized. Time dependent spatially varying forcing fields are studied in the 3:1 forced FitzHugh Nagumo system. The periodic variation of the spatially random forcing amplitude breaks the symmetry among the three quasi-homogeneous states of the system, making the three types of fronts separating phases inequivalent. The resulting inequality in the front velocities leads to the formation of "compound fronts" with velocities lying between those of the individual component fronts, and "pulses" which are analogous structures arising from the combination of three fronts. Spiral wave dynamics is studied in systems with compound fronts. (c) 2000 American Institute of Physics. PMID- 12779422 TI - Persistence of zero velocity fronts in reaction diffusion systems. AB - Steady, nonpropagating, fronts in reaction diffusion systems usually exist only for special sets of control parameters. When varying one control parameter, the front velocity may become zero only at isolated values (where the Maxwell condition is satisfied, for potential systems). The experimental observation of fronts with a zero velocity over a finite interval of parameters, e.g., in catalytic experiments [Barelko et al., Chem. Eng. Sci., 33, 805 (1978)], therefore, seems paradoxical. We show that the velocity dependence on the control parameter may be such that velocity is very small over a finite interval, and much larger outside. This happens in a class of reaction diffusion systems with two components, with the extra assumptions that (i) the two diffusion coefficients are very different, and that (ii) the slowly diffusing variables has two stable states over a control parameter range. The ratio of the two velocity scales vanishes when the smallest diffusion coefficient goes to zero. A complete study of the effect is carried out in a model of catalytic reaction. (c) 2000 American Institute of Physics. PMID- 12779423 TI - Experimental investigation of high-quality synchronization of coupled oscillators. AB - We describe two experiments in which we investigate the synchronization of coupled periodic oscillators. Each experimental system consists of two identical coupled electronic periodic oscillators that display bursts of desynchronization events similar to those observed previously in coupled chaotic systems. We measure the degree of synchronization as a function of coupling strength. In the first experiment, high-quality synchronization is achieved for all coupling strengths above a critical value. In the second experiment, no high-quality synchronization is observed. We compare our results to the predictions of the several proposed criteria for synchronization. We find that none of the criteria accurately predict the range of coupling strengths over which high-quality synchronization is observed. (c) 2000 American Institute of Physics. PMID- 12779424 TI - Embedding dynamics for round-off errors near a periodic orbit. AB - We study the propagation of round-off errors near the periodic orbits of a linear map conjugate to a planar rotation with rational rotation number. We embed the two-dimensional discrete phase space (a lattice) in a higher-dimensional torus, where points sharing the same round-off error are uniformly distributed within finitely many convex polyhedra. The embedding dynamics is linear and discontinuous, with algebraic integer coefficients. This representation affords efficient algorithms for classifying and computing the orbits and their exact densities, which we apply to the case of rational rotation number with denominator 7, corresponding to certain algebraic integers of degree three. We provide evidence that the hierarchical arrangement of orbits previously detected in quadratic cases [Lowenstein et al., Chaos 7, 49-66 (1997)] disappears, and that the growth of the number of orbits with the period is algebraic.(c) 2000 American Institute of Physics. PMID- 12779425 TI - The Sinai billiard, square torus, and field chaos. AB - An experiment is reported in which the Sinai quantum billiard and square-torus quantum billiard are compared for field chaos. In this mode of chaos, electromagnetic fields in a waveguide are analogous to the wave function. It is found that power loss in the square-torus guide exceeds that in the Sinai billiard guide by approximately 3.5 dB, thereby illustrating larger field chaos for the square-torus quantum billiard than for the Sinai quantum billiard. Solutions of the Helmholtz equation are derived for the rectangular coaxial guide that illustrate that transverse electric or transverse magnetic modes exist in the guide provided the ratio of edge lengths of the outer rectangle to parallel edge lengths of the inner rectangle is rational. Eigenfunctions partition into four sets depending on even or odd reflection properties about Cartesian axis on which the concentric rectangles are oriented. These eigenfunctions are uniquely determined by four coaxial parameters and two eigen numbers. Justification of experimental findings is based on the argument that the rationals comprise a set of measure zero with respect to the irrationals. Consequently, from an observational point of view, these modes do not exist, which is in accord with the reported experiment. (c) 2000 American Institute of Physics. PMID- 12779426 TI - N-dimensional dynamical systems exploiting instabilities in full. AB - We report experimental and numerical results showing how certain N-dimensional dynamical systems are able to exhibit complex time evolutions based on the nonlinear combination of N-1 oscillation modes. The experiments have been done with a family of thermo-optical systems of effective dynamical dimension varying from 1 to 6. The corresponding mathematical model is an N-dimensional vector field based on a scalar-valued nonlinear function of a single variable that is a linear combination of all the dynamic variables. We show how the complex evolutions appear associated with the occurrence of successive Hopf bifurcations in a saddle-node pair of fixed points up to exhaust their instability capabilities in N dimensions. For this reason the observed phenomenon is denoted as the full instability behavior of the dynamical system. The process through which the attractor responsible for the observed time evolution is formed may be rather complex and difficult to characterize. Nevertheless, the well-organized structure of the time signals suggests some generic mechanism of nonlinear mode mixing that we associate with the cluster of invariant sets emerging from the pair of fixed points and with the influence of the neighboring saddle sets on the flow nearby the attractor. The generation of invariant tori is likely during the full instability development and the global process may be considered as a generalized Landau scenario for the emergence of irregular and complex behavior through the nonlinear superposition of oscillatory motions. (c) 2000 American Institute of Physics. PMID- 12779427 TI - Anticontrol of chaos in continuous-time systems via time-delay feedback. AB - In this paper, a systematic design approach based on time-delay feedback is developed for anticontrol of chaos in a continuous-time system. This anticontrol method can drive a finite-dimensional, continuous-time, autonomous system from nonchaotic to chaotic, and can also enhance the existing chaos of an originally chaotic system. Asymptotic analysis is used to establish an approximate relationship between a time-delay differential equation and a discrete map. Anticontrol of chaos is then accomplished based on this relationship and the differential-geometry control theory. Several examples are given to verify the effectiveness of the methodology and to illustrate the systematic design procedure. (c) 2000 American Institute of Physics. PMID- 12779428 TI - Fractals and quantum mechanics. AB - A new application of a fractal concept to quantum physics has been developed. The fractional path integrals over the paths of the Levy flights are defined. It is shown that if fractality of the Brownian trajectories leads to standard quantum mechanics, then the fractality of the Levy paths leads to fractional quantum mechanics. The fractional quantum mechanics has been developed via the new fractional path integrals approach. A fractional generalization of the Schrodinger equation has been discovered. The new relationship between the energy and the momentum of the nonrelativistic fractional quantum-mechanical particle has been established, and the Levy wave packet has been introduced into quantum mechanics. The equation for the fractional plane wave function has been found. We have derived a free particle quantum-mechanical kernel using Fox's H-function. A fractional generalization of the Heisenberg uncertainty relation has been found. As physical applications of the fractional quantum mechanics we have studied a free particle in a square infinite potential well, the fractional "Bohr atom" and have developed a new fractional approach to the QCD problem of quarkonium. We also discuss the relationships between fractional and the well-known Feynman path integral approaches to quantum mechanics. (c) 2000 American Institute of Physics. PMID- 12779429 TI - Dynamical properties of chemical systems near Hopf bifurcation points. AB - In this paper, we numerically investigate local properties of dynamical systems close to a Hopf bifurcation instability. We focus on chemical systems and present an approach based on the theory of normal forms for determining numerical estimates of the limit cycle that branches off at the Hopf bifurcation point. For several numerically ill-conditioned examples taken from chemical kinetics, we compare our results with those obtained by using traditional approaches where an approximation of the limit cycle is restricted to the center subspace spanned by critical eigenvectors, and show that inclusion of higher-order terms in the normal form expansion of the limit cycle provides a significant improvement of the limit cycle estimates. This result also provides an accurate initial estimate for subsequent numerical continuation of the limit cycle. (c) 2000 American Institute of Physics. PMID- 12779430 TI - A phase transition in water coupled to a local external perturbation. AB - A flux of ideal fluid coupled to perturbation is investigated by nonperturbative methods of the quantum field theory. Asymptotic behavior of the flux coupled to perturbation turns out to be similar to that of superfluids. (c) 2000 American Institute of Physics. PMID- 12779431 TI - Asynchronous algorithm for integration of reaction-diffusion equations for inhomogeneous excitable media. AB - An asynchronous algorithm for the integration of reaction-diffusion equations for inhomogeneous excitable media is described. Since many physical systems are inhomogeneous where either the local kinetics or the diffusion or conduction properties vary significantly in space, integration schemes must be able to account for wide variations in the temporal and spatial scales of the solutions. The asynchronous algorithm utilizes a fixed spatial grid and automatically adjusts the time step locally to achieve an efficient simulation where the errors in the solution are controlled. The scheme does not depend on the specific form of the local kinetics and is easily applied to systems with complex geometries. (c) 2000 American Institute of Physics. PMID- 12779432 TI - Bistable reaction-diffusion systems can have robust zero-velocity fronts. AB - We show that for a class of bistable reaction-diffusion systems, zero-velocity fronts can be robust in the singular limit where one of the diffusion coefficients vanishes. In this case, stationary fronts can persist along variations of the system parameters. This property contrasts with the standard result that the front velocity v(&mgr;), expressed as a function of a control parameter &mgr;, is zero only at some isolated values &mgr;(0), and thus not giving robustness to zero-velocity fronts when &mgr; is varied. (c) 2000 American Institute of Physics. PMID- 12779433 TI - Nonlinear interactions in a rotating disk flow: From a Volterra model to the Ginzburg-Landau equation. AB - The physical system under consideration is the flow above a rotating disk and its cross-flow instability, which is a typical route to turbulence in three dimensional boundary layers. Our aim is to study the nonlinear properties of the wavefield through a Volterra series equation. The kernels of the Volterra expansion, which contain relevant physical information about the system, are estimated by fitting two-point measurements via a nonlinear parametric model. We then consider describing the wavefield with the complex Ginzburg-Landau equation, and derive analytical relations which express the coefficients of the Ginzburg Landau equation in terms of the kernels of the Volterra expansion. These relations must hold for a large class of weakly nonlinear systems, in fluid as well as in plasma physics. (c) 2000 American Institute of Physics. PMID- 12779434 TI - Nonlinear time series analysis of normal and pathological human walking. AB - Characterizing locomotor dynamics is essential for understanding the neuromuscular control of locomotion. In particular, quantifying dynamic stability during walking is important for assessing people who have a greater risk of falling. However, traditional biomechanical methods of defining stability have not quantified the resistance of the neuromuscular system to perturbations, suggesting that more precise definitions are required. For the present study, average maximum finite-time Lyapunov exponents were estimated to quantify the local dynamic stability of human walking kinematics. Local scaling exponents, defined as the local slopes of the correlation sum curves, were also calculated to quantify the local scaling structure of each embedded time series. Comparisons were made between overground and motorized treadmill walking in young healthy subjects and between diabetic neuropathic (NP) patients and healthy controls (CO) during overground walking. A modification of the method of surrogate data was developed to examine the stochastic nature of the fluctuations overlying the nominally periodic patterns in these data sets. Results demonstrated that having subjects walk on a motorized treadmill artificially stabilized their natural locomotor kinematics by small but statistically significant amounts. Furthermore, a paradox previously present in the biomechanical literature that resulted from mistakenly equating variability with dynamic stability was resolved. By slowing their self-selected walking speeds, NP patients adopted more locally stable gait patterns, even though they simultaneously exhibited greater kinematic variability than CO subjects. Additionally, the loss of peripheral sensation in NP patients was associated with statistically significant differences in the local scaling structure of their walking kinematics at those length scales where it was anticipated that sensory feedback would play the greatest role. Lastly, stride-to stride fluctuations in the walking patterns of all three subject groups were clearly distinguishable from linearly autocorrelated Gaussian noise. As a collateral benefit of the methodological approach taken in this study, some of the first steps at characterizing the underlying structure of human locomotor dynamics have been taken. Implications for understanding the neuromuscular control of locomotion are discussed. (c) 2000 American Institute of Physics. PMID- 12779435 TI - Timely detection of dynamical change in scalp EEG signals. AB - We present a robust, model-independent technique for quantifying changes in the dynamics underlying nonlinear time-serial data. After constructing discrete density distributions of phase-space points on the attractor for time-windowed data sets, we measure the dissimilarity between density distributions via L(1) distance and chi(2) statistics. The discriminating power of the new measures is first tested on data generated by the Bondarenko "synthetic brain" model. We also compare traditional nonlinear measures and the new dissimilarity measures to detect dynamical change in scalp EEG data. The results demonstrate a clear superiority of the new measures in comparison to traditional nonlinear measures as robust and timely discriminators of changing dynamics. (c) 2000 American Institute of Physics. PMID- 12779436 TI - Exact solutions to chaotic and stochastic systems. AB - We investigate functions that are exact solutions to chaotic dynamical systems. A generalization of these functions can produce truly random numbers. For the first time, we present solutions to random maps. This allows us to check, analytically, some recent results about the complexity of random dynamical systems. We confirm the result that a negative Lyapunov exponent does not imply predictability in random systems. We test the effectiveness of forecasting methods in distinguishing between chaotic and random time series. Using the explicit random functions, we can give explicit analytical formulas for the output signal in some systems with stochastic resonance. We study the influence of chaos on the stochastic resonance. We show, theoretically, the existence of a new type of solitonic stochastic resonance, where the shape of the kink is crucial. Using our models we can predict specific patterns in the output signal of stochastic resonance systems. (c) 2001 American Institute of Physics. PMID- 12779437 TI - Geometrical constraints on finite-time Lyapunov exponents in two and three dimensions. AB - Constraints are found on the spatial variation of finite-time Lyapunov exponents of two- and three-dimensional systems of ordinary differential equations. In a chaotic system, finite-time Lyapunov exponents describe the average rate of separation, along characteristic directions, of neighboring trajectories. The solution of the equations is a coordinate transformation that takes initial conditions (the Lagrangian coordinates) to the state of the system at a later time (the Eulerian coordinates). This coordinate transformation naturally defines a metric tensor, from which the Lyapunov exponents and characteristic directions are obtained. By requiring that the Riemann curvature tensor vanish for the metric tensor (a basic result of differential geometry in a flat space), differential constraints relating the finite-time Lyapunov exponents to the characteristic directions are derived. These constraints are realized with exponential accuracy in time. A consequence of the relations is that the finite time Lyapunov exponents are locally small in regions where the curvature of the stable manifold is large, which has implications for the efficiency of chaotic mixing in the advection-diffusion equation. The constraints also modify previous estimates of the asymptotic growth rates of quantities in the dynamo problem, such as the magnitude of the induced current. (c) 2001 American Institute of Physics. PMID- 12779438 TI - Chaotic synchronization of coupled ergodic maps. AB - With few exceptions, studies of chaotic synchronization have focused on dissipative chaos. Though less well known, chaotic systems that lack dissipation may also synchronize. Motivated by an application in communication systems, we couple a family of ergodic maps on the N-torus and study the global stability of the synchronous state. While most trajectories synchronize at some time, there is a measure zero set that never synchronizes. We give explicit examples of these asynchronous orbits in dimensions two and four. On more typical trajectories, the synchronization error reaches arbitrarily small values and, in practice, converges. In dimension two we derive bounds on the average synchronization time for trajectories resulting from randomly chosen initial conditions. Numerical experiments suggest similar bounds exist in higher dimensions as well. Adding noise to the coupling signal destroys the invariance of the synchronous state and causes typical trajectories to desynchronize. We propose a modification of the standard coupling scheme that corrects this problem resulting in robust synchronization in the presence of noise. PMID- 12779439 TI - Synchronization experiments with an atmospheric global circulation model. AB - Synchronization in a chaotic system with many degrees of freedom is investigated by coupling two identical global atmospheric circulation models. Starting from different initial conditions, the two submodels show complete synchronization as well as noncomplete synchronization depending on the coupling strength. The relatively low value of the coupling strength threshold for complete synchronization indicates the potential importance of synchronization mechanisms involved in climate variability. In addition, the results suggest synchronization experiments as a valuable additional method to analyze complex dynamical models, e.g., to estimate the largest Lyapunov exponent. (c) 2001 American Institute of Physics. PMID- 12779440 TI - Parametrically forced pattern formation. AB - Pattern formation in a nonlinear damped Mathieu-type partial differential equation defined on one space variable is analyzed. A bifurcation analysis of an averaged equation is performed and compared to full numerical simulations. Parametric resonance leads to periodically varying patterns whose spatial structure is determined by amplitude and detuning of the periodic forcing. At onset, patterns appear subcritically and attractor crowding is observed for large detuning. The evolution of patterns under the increase of the forcing amplitude is studied. It is found that spatially homogeneous and temporally periodic solutions occur for all detuning at a certain amplitude of the forcing. Although the system is dissipative, spatial solitons are found representing domain walls creating a phase jump of the solutions. Qualitative comparisons with experiments in vertically vibrating granular media are made. (c) 2001 American Institute of Physics. PMID- 12779441 TI - Dynamics of spatially nonuniform patterning in the model of blood coagulation. AB - We propose a reaction-diffusion model that describes in detail the cascade of molecular events during blood coagulation. In a reduced form, this model contains three equations in three variables, two of which are self-accelerated. One of these variables, an activator, behaves in a threshold manner. An inhibitor is also produced autocatalytically, but there is no inhibitor threshold, because it is generated only in the presence of the activator. All model variables are set to have equal diffusion coefficients. The model has a stable stationary trivial state, which is spatially uniform and an excitation threshold. A pulse of excitation runs from the point where the excitation threshold has been exceeded. The regime of its propagation depends on the model parameters. In a one dimensional problem, the pulse either stops running at a certain distance from the excitation point, or it reaches the boundaries as an autowave. However, there is a parameter range where the pulse does not disappear after stopping and exists stationarily. The resulting steady-state profiles of the model variables are symmetrical relative to the center of the structure formed. (c) 2001 American Institute of Physics. PMID- 12779442 TI - Filament instability and rotational tissue anisotropy: A numerical study using detailed cardiac models. AB - The role of cardiac tissue anisotropy in the breakup of vortex filaments is studied using two detailed cardiac models. In the Beeler-Reuter model, modified to produce stable spiral waves in two dimensions, we find that anisotropy can destabilize a vortex filament in a parallelepipedal slab of tissue. The mechanisms of the instability are similar to the ones reported in previous work on a simplified cardiac model by Fenton and Karma [Chaos 8, 20 (1998)]. In the Luo-Rudy model, also modified to produce stable spiral waves in two dimensions, we find that anisotropy does not destabilize filaments. A possible explanation for this model-dependent behavior based on spiral tip trajectories is offered. (c) 2001 American Institute of Physics. PMID- 12779443 TI - Molecular, metabolic, and genetic control: An introduction. AB - The living cell is a miniature, self-reproducing, biochemical machine. Like all machines, it has a power supply, a set of working components that carry out its necessary tasks, and control systems that ensure the proper coordination of these tasks. In this Special Issue, we focus on the molecular regulatory systems that control cell metabolism, gene expression, environmental responses, development, and reproduction. As for the control systems in human-engineered machines, these regulatory networks can be described by nonlinear dynamical equations, for example, ordinary differential equations, reaction-diffusion equations, stochastic differential equations, or cellular automata. The articles collected here illustrate (i) a range of theoretical problems presented by modern concepts of cellular regulation, (ii) some strategies for converting molecular mechanisms into dynamical systems, (iii) some useful mathematical tools for analyzing and simulating these systems, and (iv) the sort of results that derive from serious interplay between theory and experiment. (c) 2001 American Institute of Physics. PMID- 12779444 TI - Molecular interaction maps as information organizers and simulation guides. AB - A graphical method for mapping bioregulatory networks is presented that is suited for the representation of multimolecular complexes, protein modifications, as well as actions at cell membranes and between protein domains. The symbol conventions defined for these molecular interaction maps are designed to accommodate multiprotein assemblies and protein modifications that can generate combinatorially large numbers of molecular species. Diagrams can either be "heuristic," meaning that detailed knowledge of all possible reaction paths is not required, or "explicit," meaning that the diagrams are totally unambiguous and suitable for simulation. Interaction maps are linked to annotation lists and indexes that provide ready access to pertinent data and references, and that allow any molecular species to be easily located. Illustrative interaction maps are included on the domain interactions of Src, transcription control of E2F regulated genes, and signaling from receptor tyrosine kinase through phosphoinositides to Akt/PKB. A simple method of going from an explicit interaction diagram to an input file for a simulation program is outlined, in which the differential equations need not be written out. The role of interaction maps in selecting and defining systems for modeling is discussed. (c) 2001 American Institute of Physics. PMID- 12779445 TI - Extracting information from cDNA arrays. AB - High-density DNA arrays allow measurements of gene expression levels (messenger RNA abundance) for thousands of genes simultaneously. We analyze arrays with spotted cDNA used in monitoring of expression profiles. A dilution series of a mouse liver probe is deployed to quantify the reproducibility of expression measurements. Saturation effects limit the accessible signal range at high intensities. Additive noise and outshining from neighboring spots dominate at low intensities. For repeated measurements on the same filter and filter-to-filter comparisons correlation coefficients of 0.98 are found. Next we consider the clustering of gene expression time series from stimulated human fibroblasts which aims at finding co-regulated genes. We analyze how preprocessing, the distance measure, and the clustering algorithm affect the resulting clusters. Finally we discuss algorithms for the identification of transcription factor binding sites from clusters of co-regulated genes. (c) 2001 American Institute of Physics. PMID- 12779446 TI - On the deduction of chemical reaction pathways from measurements of time series of concentrations. AB - We discuss the deduction of reaction pathways in complex chemical systems from measurements of time series of chemical concentrations of reacting species. First we review a technique called correlation metric construction (CMC) and show the construction of a reaction pathway from measurements on a part of glycolysis. Then we present two new improved methods for the analysis of time series of concentrations, entropy metric construction (EMC), and entropy reduction method (ERM), and illustrate (EMC) with calculations on a model reaction system. (c) 2001 American Institute of Physics. PMID- 12779447 TI - Analysis of nonlinear dynamics on arbitrary geometries with the Virtual Cell. AB - The Virtual Cell is a modeling tool that allows biologists and theorists alike to specify and simulate cell-biophysical models on arbitrarily complex geometries. The framework combines an intuitive, front-end graphical user interface that runs in a web browser, sophisticated server-side numerical algorithms, a database for storage of models and simulation results, and flexible visualization capabilities. In this paper, we present an overview of the capabilities of the Virtual Cell, and, for the first time, the detailed mathematical formulation used as the basis for spatial computations. We also present summaries of two rather typical modeling projects, in order to illustrate the principal capabilities of the Virtual Cell. (c) 2001 American Institute of Physics. PMID- 12779449 TI - Design principles for elementary gene circuits: Elements, methods, and examples. AB - The control of gene expression involves complex circuits that exhibit enormous variation in design. For years the most convenient explanation for these variations was historical accident. According to this view, evolution is a haphazard process in which many different designs are generated by chance; there are many ways to accomplish the same thing, and so no further meaning can be attached to such different but equivalent designs. In recent years a more satisfying explanation based on design principles has been found for at least certain aspects of gene circuitry. By design principle we mean a rule that characterizes some biological feature exhibited by a class of systems such that discovery of the rule allows one not only to understand known instances but also to predict new instances within the class. The central importance of gene regulation in modern molecular biology provides strong motivation to search for more of these underlying design principles. The search is in its infancy and there are undoubtedly many design principles that remain to be discovered. The focus of this three-part review will be the class of elementary gene circuits in bacteria. The first part reviews several elements of design that enter into the characterization of elementary gene circuits in prokaryotic organisms. Each of these elements exhibits a variety of realizations whose meaning is generally unclear. The second part reviews mathematical methods used to represent, analyze, and compare alternative designs. Emphasis is placed on particular methods that have been used successfully to identify design principles for elementary gene circuits. The third part reviews four design principles that make specific predictions regarding (1) two alternative modes of gene control, (2) three patterns of coupling gene expression in elementary circuits, (3) two types of switches in inducible gene circuits, and (4) the realizability of alternative gene circuits and their response to phased environmental cues. In each case, the predictions are supported by experimental evidence. These results are important for understanding the function, design, and evolution of elementary gene circuits. (c) 2001 American Institute of Physics. PMID- 12779448 TI - How gap genes make their domains: An analytical study based on data driven approximations. AB - We consider a mathematical formulation of the problem of protein production during segment determination in the Drosophila blastoderm, together with some preliminary results of its analytical study. We reformulate the spatial difference equations as a set of nonlinear partial differential equations and obtain their dimensionless form in the continuum limit. Using previous results obtained by the gene circuit method, we find an asymptotic statement of the problem with a small parameter. Some results of the comparison method applied to the model are obtained, and exact stationary upper solutions are derived. They exhibit distinctive features of localized bell-shaped structures. (c) 2001 American Institute of Physics. PMID- 12779450 TI - Symbolic dynamics and computation in model gene networks. AB - We analyze a class of ordinary differential equations representing a simplified model of a genetic network. In this network, the model genes control the production rates of other genes by a logical function. The dynamics in these equations are represented by a directed graph on an n-dimensional hypercube (n cube) in which each edge is directed in a unique orientation. The vertices of the n-cube correspond to orthants of state space, and the edges correspond to boundaries between adjacent orthants. The dynamics in these equations can be represented symbolically. Starting from a point on the boundary between neighboring orthants, the equation is integrated until the boundary is crossed for a second time. Each different cycle, corresponding to a different sequence of orthants that are traversed during the integration of the equation always starting on a boundary and ending the first time that same boundary is reached, generates a different letter of the alphabet. A word consists of a sequence of letters corresponding to a possible sequence of orthants that arise from integration of the equation starting and ending on the same boundary. The union of the words defines the language. Letters and words correspond to analytically computable Poincare maps of the equation. This formalism allows us to define bifurcations of chaotic dynamics of the differential equation that correspond to changes in the associated language. Qualitative knowledge about the dynamics found by integrating the equation can be used to help solve the inverse problem of determining the underlying network generating the dynamics. This work places the study of dynamics in genetic networks in a context comprising both nonlinear dynamics and the theory of computation. (c) 2001 American Institute of Physics. PMID- 12779451 TI - Multistationarity, the basis of cell differentiation and memory. I. Structural conditions of multistationarity and other nontrivial behavior. AB - A biological introduction serves to remind us that differentiation is an epigenetic process, that multistationarity can account for epigenetic differences, including those involved in cell differentiation, and that positive feedback circuits are a necessary condition for multistationarity and, by inference, for differentiation. The core of the paper is comprised of a formal description of feedback circuits and unions of disjoint circuits. We introduce the concepts of full-circuit (a circuit or union of disjoint circuits which involves all the variables of the system), and of ambiguous circuit (a circuit whose sign depends on the location in phase space). We describe the partition of phase space (a) according to the signs of the ambiguous circuits, and (b) according to the signs of the eigenvalues or their real part. We introduce a normalization of the system versus one of the circuits; in two variables, this permits an entirely general description in terms of a common diagram in the "circuit space." The paper ends with general statements concerning the requirements for multistationarity, stable periodicity, and deterministic chaos. (c) 2001 American Institute of Physics. PMID- 12779452 TI - Multistationarity, the basis of cell differentiation and memory. II. Logical analysis of regulatory networks in terms of feedback circuits. AB - Circuits and their involvement in complex dynamics are described in differential terms in Part I of this work. Here, we first explain why it may be appropriate to use a logical description, either by itself or in symbiosis with the differential description. The major problem of a logical description is to find an adequate way to involve time. The procedure we adopted differs radically from the classical one by its fully asynchronous character. In Sec. II we describe our "naive" logical approach, and use it to illustrate the major laws of circuitry (namely, the involvement of positive circuits in multistationarity and of negative circuits in periodicity) and in a biological example. Already in the naive description, the major steps of the logical description are to: (i) describe a model as a set of logical equations, (ii) derive the state table from the equations, (iii) derive the graph of the sequences of states from the state table, and (iv) determine which of the possible pathways will be actually followed in terms of time delays. In the following sections we consider multivalued variables where required, the introduction of logical parameters and of logical values ascribed to the thresholds, and the concept of characteristic state of a circuit. This generalized logical description provides an image whose qualitative fit with the differential description is quite remarkable. A major interest of the generalized logical description is that it implies a limited and often quite small number of possible combinations of values of the logical parameters. The space of the logical parameters is thus cut into a limited number of boxes, each of which is characterized by a defined qualitative behavior of the system. Our analysis tells which constraints on the logical parameters must be fulfilled in order for any circuit (or combination of circuits) to be functional. Functionality of a circuit will result in multistationarity (in the case of a positive circuit) or in a cycle (in the case of a negative circuit). The last sections deal with "more about time delays" and "reverse logic," an approach that aims to proceed rationally from facts to models. (c) 2001 American Institute of Physics. PMID- 12779453 TI - Invariant manifold methods for metabolic model reduction. AB - After the decay of transients, the behavior of a set of differential equations modeling a chemical or biochemical system generally rests on a low-dimensional surface which is an invariant manifold of the flow. If an equation for such a manifold can be obtained, the model has effectively been reduced to a smaller system of differential equations. Using perturbation methods, we show that the distinction between rapidly decaying and long-lived (slow) modes has a rigorous basis. We show how equations for attracting invariant (slow) manifolds can be constructed by a geometric approach based on functional equations derived directly from the differential equations. We apply these methods to two simple metabolic models. (c) 2001 American Institute of Physics. PMID- 12779454 TI - Designer gene networks: Towards fundamental cellular control. AB - The engineered control of cellular function through the design of synthetic genetic networks is becoming plausible. Here we show how a naturally occurring network can be used as a parts list for artificial network design, and how model formulation leads to computational and analytical approaches relevant to nonlinear dynamics and statistical physics. We first review the relevant work on synthetic gene networks, highlighting the important experimental findings with regard to genetic switches and oscillators. We then present the derivation of a deterministic model describing the temporal evolution of the concentration of protein in a single-gene network. Bistability in the steady-state protein concentration arises naturally as a consequence of autoregulatory feedback, and we focus on the hysteretic properties of the protein concentration as a function of the degradation rate. We then formulate the effect of an external noise source which interacts with the protein degradation rate. We demonstrate the utility of such a formulation by constructing a protein switch, whereby external noise pulses are used to switch the protein concentration between two values. Following the lead of earlier work, we show how the addition of a second network component can be used to construct a relaxation oscillator, whereby the system is driven around the hysteresis loop. We highlight the frequency dependence on the tunable parameter values, and discuss design plausibility. We emphasize how the model equations can be used to develop design criteria for robust oscillations, and illustrate this point with parameter plots illuminating the oscillatory regions for given parameter values. We then turn to the utilization of an intrinsic cellular process as a means of controlling the oscillations. We consider a network design which exhibits self-sustained oscillations, and discuss the driving of the oscillator in the context of synchronization. Then, as a second design, we consider a synthetic network with parameter values near, but outside, the oscillatory boundary. In this case, we show how resonance can lead to the induction of oscillations and amplification of a cellular signal. Finally, we construct a toggle switch from positive regulatory elements, and compare the switching properties for this network with those of a network constructed using negative regulation. Our results demonstrate the utility of model analysis in the construction of synthetic gene regulatory networks. (c) 2001 American Institute of Physics. PMID- 12779455 TI - Robustness of the bistable behavior of a biological signaling feedback loop. AB - Biological signaling networks comprised of cellular components including signaling proteins and small molecule messengers control the many cell function in responses to various extracellular and intracellular signals including hormone and neurotransmitter inputs, and genetic events. Many signaling pathways have motifs familiar to electronics and control theory design. Feedback loops are among the most common of these. Using experimentally derived parameters, we modeled a positive feedback loop in signaling pathways used by growth factors to trigger cell proliferation. This feedback loop is bistable under physiological conditions, although the system can move to a monostable state as well. We find that bistability persists under a wide range of regulatory conditions, even when core enzymes in the feedback loop deviate from physiological values. We did not observe any other phenomena in the core feedback loop, but the addition of a delayed inhibitory feedback was able to generate oscillations under rather extreme parameter conditions. Such oscillations may not be of physiological relevance. We propose that the kinetic properties of this feedback loop have evolved to support bistability and flexibility in going between bistable and monostable modes, while simultaneously being very refractory to oscillatory states. (c) 2001 American Institute of Physics. PMID- 12779456 TI - Bistability in cell signaling: How to make continuous processes discontinuous, and reversible processes irreversible. AB - Xenopus oocyte maturation is an example of an all-or-none, irreversible cell fate induction process. In response to a submaximal concentration of the steroid hormone progesterone, a given oocyte may either mature or not mature, but it can exist in intermediate states only transiently. Moreover, once an oocyte has matured, it will remain arrested in the mature state even after the progesterone is removed. It has been hypothesized that the all-or-none character of oocyte maturation, and some aspects of the irreversibility of maturation, arise out of the bistability of the signal transduction system that triggers maturation. The bistability, in turn, is hypothesized to arise from the way the signal transducers are organized into a signaling circuit that includes positive feedback (which makes it so that the system cannot rest in intermediate states) and ultrasensitivity (which filters small stimuli out of the feedback loop, allowing the system to have a stable off-state). Here we review two simple graphical methods that are commonly used to analyze bistable systems, discuss the experimental evidence for bistability in oocyte maturation, and suggest that bistability may be a common means of producing all-or-none responses and a type of biochemical memory. (c) 2001 American Institute of Physics. PMID- 12779457 TI - A bifurcation analysis of two coupled calcium oscillators. AB - In many cell types, asynchronous or synchronous oscillations in the concentration of intracellular free calcium occur in adjacent cells that are coupled by gap junctions. Such oscillations are believed to underlie oscillatory intercellular calcium waves in some cell types, and thus it is important to understand how they occur and are modified by intercellular coupling. Using a previous model of intracellular calcium oscillations in pancreatic acinar cells, this article explores the effects of coupling two cells with a simple linear diffusion term. Depending on the concentration of a signal molecule, inositol (1,4,5) trisphosphate, coupling two identical cells by diffusion can give rise to synchronized in-phase oscillations, as well as different-amplitude in-phase oscillations and same-amplitude antiphase oscillations. Coupling two nonidentical cells leads to more complex behaviors such as cascades of period doubling and multiply periodic solutions. This study is a first step towards understanding the role and significance of the diffusion of calcium through gap junctions in the coordination of oscillatory calcium waves in a variety of cell types. (c) 2001 American Institute of Physics. PMID- 12779459 TI - Dynamic behavior in mathematical models of the tryptophan operon. AB - This paper surveys the general theory of operon regulation as first formulated by Goodwin and Griffith, and then goes on to consider in detail models of regulation of tryptophan production by Bliss, Sinha, and Santillan and Mackey, and the interrelationships between them. We further give a linear stability analysis of the Santillan and Mackey model for wild type E. coli as well as three different mutant strains that have been previously studied in the literature. This stability analysis indicates that the tryptophan production systems should be stable, which is in accord with our numerical results. (c) 2001 American Institute of Physics. PMID- 12779458 TI - From simple to complex oscillatory behavior in metabolic and genetic control networks. AB - We present an overview of mechanisms responsible for simple or complex oscillatory behavior in metabolic and genetic control networks. Besides simple periodic behavior corresponding to the evolution toward a limit cycle we consider complex modes of oscillatory behavior such as complex periodic oscillations of the bursting type and chaos. Multiple attractors are also discussed, e.g., the coexistence between a stable steady state and a stable limit cycle (hard excitation), or the coexistence between two simultaneously stable limit cycles (birhythmicity). We discuss mechanisms responsible for the transition from simple to complex oscillatory behavior by means of a number of models serving as selected examples. The models were originally proposed to account for simple periodic oscillations observed experimentally at the cellular level in a variety of biological systems. In a second stage, these models were modified to allow for complex oscillatory phenomena such as bursting, birhythmicity, or chaos. We consider successively (1) models based on enzyme regulation, proposed for glycolytic oscillations and for the control of successive phases of the cell cycle, respectively; (2) a model for intracellular Ca(2+) oscillations based on transport regulation; (3) a model for oscillations of cyclic AMP based on receptor desensitization in Dictyostelium cells; and (4) a model based on genetic regulation for circadian rhythms in Drosophila. Two main classes of mechanism leading from simple to complex oscillatory behavior are identified, namely (i) the interplay between two endogenous oscillatory mechanisms, which can take multiple forms, overt or more subtle, depending on whether the two oscillators each involve their own regulatory feedback loop or share a common feedback loop while differing by some related process, and (ii) self-modulation of the oscillator through feedback from the system's output on one of the parameters controlling oscillatory behavior. However, the latter mechanism may also be viewed as involving the interplay between two feedback processes, each of which might be capable of producing oscillations. Although our discussion primarily focuses on the case of autonomous oscillatory behavior, we also consider the case of nonautonomous complex oscillations in a model for circadian oscillations subjected to periodic forcing by a light-dark cycle and show that the occurrence of entrainment versus chaos in these conditions markedly depends on the wave form of periodic forcing. (c) 2001 American Institute of Physics. PMID- 12779460 TI - Kick-starting the cell cycle: From growth-factor stimulation to initiation of DNA replication. AB - The essential genes, proteins and associated regulatory networks involved in the entry into the mammalian cell cycle are identified, from activation of growth factor receptors to intracellular signal transduction pathways that impinge on the cell cycle machinery and ultimately on the initiation of DNA replication. Signaling pathways mediated by the oncoproteins Ras and Myc induce the activation of cyclin-dependent kinases CDK4 and CDK2, and the assembly and firing of pre replication complexes require a collaboration among E2F, CDK2, and Cdc7 kinase. A proposed core mechanism of the restriction point, the major checkpoint prior to commitment to DNA synthesis, involves cyclin E/CDK2, the phosphatase Cdc25A, and the CDK inhibitor p27Kip1. (c) 2001 American Institute of Physics. PMID- 12779461 TI - Mathematical model of the cell division cycle of fission yeast. AB - Much is known about the genes and proteins controlling the cell cycle of fission yeast. Can these molecular components be spun together into a consistent mechanism that accounts for the observed behavior of growth and division in fission yeast cells? To answer this question, we propose a mechanism for the control system, convert it into a set of 14 differential and algebraic equations, study these equations by numerical simulation and bifurcation theory, and compare our results to the physiology of wild-type and mutant cells. In wild-type cells, progress through the cell cycle (G1-->S-->G2-->M) is related to cyclic progression around a hysteresis loop, driven by cell growth and chromosome alignment on the metaphase plate. However, the control system operates much differently in double-mutant cells, wee1(-) cdc25Delta, which are defective in progress through the latter half of the cell cycle (G2 and M phases). These cells exhibit "quantized" cycles (interdivision times clustering around 90, 160, and 230 min). We show that these quantized cycles are associated with a supercritical Hopf bifurcation in the mechanism, when the wee1 and cdc25 genes are disabled. (c) 2001 American Institute of Physics. PMID- 12779462 TI - Topographic organization of Hebbian neural connections by synchronous wave activity. AB - Experimental studies have revealed that the refinement of early, imprecise connections in the developing visual system involves activity in the retina before the onset of vision. We study the evolution of initially random unidirectional connections between two excitable layers of FitzHugh-Nagumo neurons with simulated spontaneous activity in the input layer. Lateral coupling within the layers yields synchronous neural wave activity that serves as a template for the Hebbian learning process, which establishes topographically precise interlayer connections. (c) 2001 American Institute of Physics. PMID- 12779463 TI - Weak mixing and anomalous kinetics along filamented surfaces. AB - We consider chaotic properties of a particle in a square billiard with a horizontal bar in the middle. Such a system can model field-line windings of the merged surfaces. The system has weak-mixing properties with zero Lyapunov exponent and entropy, and it can be also interesting as an example of a system with intermediate chaotic properties, between the integrability and strong mixing. We show that the transport is anomalous and that its properties can be linked to the ergodic properties of continued fractions. The distribution of Poincare recurrences, distribution of the displacements, and the moments of the truncated distribution of the displacements are obtained. Connections between different exponents are found. It is shown that the distribution function of displacements and its truncated moments as a function of time exhibit log periodic oscillations (modulations) with a universal period T(log)=pi(2)/12 ln 2. We note that similar results are valid for a family of billiard, particularly for billiards with square-in-square geometry. (c) 2001 American Institute of Physics. PMID- 12779464 TI - Lowering the dimension of polynomial vector fields in R(2) and R(3). AB - We classify the normal forms associated to polynomial vector fields with dimensions two and three whose principal part is linear. Then we reduce by one the dimension of the associated differential systems. This is achieved by means of the extension of a symmetry of the unperturbed part to the whole system, up to a certain order of approximation. The corresponding transformation is formal in the sense that we have no estimation on its radius of convergence. We calculate the invariants, reduced phase spaces and differential systems associated to each reduction. This is the general setting for the study of the dynamics of the reduced system in order to obtain information about the original one. (c) 2001 American Institute of Physics. PMID- 12779465 TI - Efficient noncausal noise reduction for deterministic time series. AB - We present a simple noncausal noise reduction algorithm for time series that consist of noisy measurements of the state vectors of a deterministic (chaotic) nonlinear system. The underlying dynamical system is assumed to be known and to operate in discrete time. The noise reduction algorithm is an iterative scheme for finding exact deterministic orbits close to the measured noisy orbits. Furthermore, we discuss cases where the solution is not the original orbit but homoclinic to it. (c) 2001 American Institute of Physics. PMID- 12779466 TI - Orbits in the H(2)O molecule. AB - We study the forms of the orbits in a symmetric configuration of a realistic model of the H(2)O molecule with particular emphasis on the periodic orbits. We use an appropriate Poincare surface of section (PSS) and study the distribution of the orbits on this PSS for various energies. We find both ordered and chaotic orbits. The proportion of ordered orbits is almost 100% for small energies, but decreases abruptly beyond a critical energy. When the energy exceeds the escape energy there are still nonescaping orbits around stable periodic orbits. We study in detail the forms of the various periodic orbits, and their connections, by providing appropriate stability and bifurcation diagrams. (c) 2001 American Institute of Physics. PMID- 12779467 TI - Coupled catalytic oscillators: Beyond the mass-action law. AB - We present Monte Carlo simulations of the reaction kinetics corresponding to two coupled catalytic oscillators in the case when oscillations result from the interplay between the reaction steps and adsorbate-induced surface restructuring. The model used is aimed to mimic oscillations on a single nm catalyst particle with two kinds of facets or on two catalyst particles on a support. Specifically, we treat the NO reduction by H(2) on a composite catalyst containing two catalytically active Pt(100) parts connected by an inactive link. The catalyst is represented by a rectangular fragment of a square lattice. The left- and right hand parts of the lattice mimic Pt(100). With an appropriate choice of the model parameters, these sublattices play a role of catalytic oscillators. The central catalytically inactive sublattice is considered to be able only to adsorb NO reversibly and can be viewed as a Pt(111) facet or a support. The interplay of the reactions running on the catalytically active areas occurs via NO diffusion over the boundaries between the sublattices. Using this model, we show that the coupling of the catalytically active sublattices may synchronize nearly harmonic oscillations observed on these sublattices and also may result in the appearance of aperiodic partly synchronized oscillations. The spatio-temporal patterns corresponding to these regimes are nontrivial. In particular, the model predicts that, due to phase separation, the reaction may be accompanied by the formation of narrow NO-covered zones on the left and right sublattices near the boundaries between these sublattices and the central sublattice. Such patterns cannot be obtained by using the conventional mean-field reaction-diffusion equations based on the mass-action law. The experimental opportunities to observe the predicted phenomena are briefly discussed. (c) 2001 American Institute of Physics. PMID- 12779468 TI - Two models for the dynamics of boiling in a short capillary tube. AB - We investigate the dynamical origin of the statistical properties of boiling in a short capillary tube. Two different models are proposed (Markov model and rotational model). We show that the behavior of each model may be related to different properties of the physical phenomenon. We conclude with a suggestion of a new experimental measurement which can help to adjust several aspects of the models. (c) 2001 American Institute of Physics. PMID- 12779469 TI - Particle dynamics and mixing in the frequency driven "Kelvin cat eyes" flow. AB - The "Kelvin cat eyes" stream function is used as a simple fluid flow model to study particle dynamics, mixing and transport in a two-dimensional time-dependent flow field. Lagrangian formulation is used to describe the motion of small spherical particles present in the flow. Individual particle trajectories, under the influence of various flow parameters are studied. The equation describing the motion of these particles constitutes a set of first-order nonlinear differential equations describing a dynamical system. The time-dependent Eulerian flow field is studied as a nonintegrable Hamiltonian system in order to get insight into the underlying nonlinear properties of the system, which directly influence its complicated transport and mixing behavior. Chaotic advection (Lagrangian turbulence) was observed for heavy particles (high Stokes numbers) while no stochastic behavior was observed for light particles. The introduction of perturbation had only a limited effect on individual particle trajectories. However, the introduction of perturbation caused a shrinking of the phase space where bounded stochastic or quasi-periodic motion occurs. This phenomenon can lead to a better understanding of the link between the behavior of the underlying flow in the Hamiltonian formulation and the dynamics of the passive scalars in the Lagrangian description. The Eulerian flow field itself was found to behave chaotically under the influence of a periodic perturbation, because the stable and unstable manifolds associated with neighboring hyperbolic points intersected. This coincides with the better mixing of the fluid. Stochasticity was also discovered close to the periodic points of the system using Poincare maps. Mixing and transport properties are analyzed as a function of the perturbation frequency. (c) 2001 American Institute of Physics. PMID- 12779470 TI - Julia sets for the super-Newton method, Cauchy's method, and Halley's method. AB - We study numerically and dynamically three cubically convergent iterative root finding algorithms, namely Cauchy's method, the super-Newton method, and Halley's method. Using the concept of a universal Julia set (motivated by the results of McMullen), we establish that these algorithms converge when applied to any quadratic with distinct roots. We give examples showing the existence of attracting periodic orbits not associated to a root for the super-Newton method and Halley's method applied to cubic polynomials. We include computer plots showing the dynamic structure for each algorithm applied to a variety of polynomials. (c) 2001 American Institute of Physics. PMID- 12779471 TI - Influence of low intensity noise on assemblies of diffusively coupled chaotic cells. AB - The effect of time-correlated and white Gaussian noises of low intensity on one dimensional arrays consisting of diffusively coupled chaotic cells is analyzed. An improvement or worsening of the synchronization between cells of the array driven by low-intensity colored noise is observed for a resonant interval of time correlation values. A comparison between colored and white noise and additive and multiplicative contribution has been carried out investigating the nonlinear cooperative effects of noise strength, correlation time, and coupling strength to control spatiotemporal chaos in coupled arrays of chaotic cells. The possibility to distinguish highly correlated areas of a diffusively coupled network of cells by using low-intensity time correlated noise is discussed. (c) 2001 American Institute of Physics. PMID- 12779472 TI - The benefits of multibit chaotic sigma delta modulation. AB - Sigma delta modulation is a popular technique for high-resolution analog-to digital conversion and digital-to-analog conversion. We investigate chaotic phenomena in multibit first-order sigma-delta modulators. Particular attention is placed on the occurrence of periodic orbits or limit cycles. These may result in idle tones audible to the listener when sigma-delta modulation is used for audio signal processing. One suggested method of eliminating idle tones is the operation of a sigma delta modulator in the chaotic regime. Unfortunately, chaotic modulation of a first order sigma delta modulator is a poor system for signal processing. We show that minor variations on a traditional first order sigma-delta modulator, together with a multibit implementation, may be used to produce an effective, stable chaotic modulator that accurately encodes the input and helps remove the presence of idle tones. (c) 2001 American Institute of Physics. PMID- 12779473 TI - Directional fractional kinetics. AB - Kinetic equations used to describe systems with dynamical chaos may contain fractional derivatives of an order alpha in space and beta in time in order to represent processes of stickiness, intermittency, and so on. We demonstrate for a simple example that the kinetics is anisotropic not only in the angular dependence of the diffusion constant, but also in the angular dependence of the exponents alpha and beta. A theory of such kinetic processes has been developed on the basis of integral representation and asymptotic solutions for different cases have been obtained. The results show the existence of self-similar solutions as well as possible logarithmic deviations. (c) 2001 American Institute of Physics. PMID- 12779474 TI - Population dynamics advected by chaotic flows: A discrete-time map approach. AB - A discrete-time model of reacting evolving fields, transported by a bidimensional chaotic fluid flow, is studied. Our approach is based on the use of a Lagrangian scheme where fluid particles are advected by a two-dimensional symplectic map possibly yielding Lagrangian chaos. Each fluid particle carries concentrations of active substances which evolve according to its own reaction dynamics. This evolution is also modeled in terms of maps. Motivated by the question, of relevance in marine ecology, of how a localized distribution of nutrients or preys affects the spatial structure of predators transported by a fluid flow, we study a specific model in which the population dynamics is given by a logistic map with space-dependent coefficient, and advection is given by the standard map. Fractal and random patterns in the Eulerian spatial concentration of predators are obtained under different conditions. Exploiting the analogies of this coupled map (advection plus reaction) system with a random map, some features of these patterns are discussed. (c) 2001 American Institute of Physics. PMID- 12779475 TI - In-out intermittency in partial differential equation and ordinary differential equation models. AB - We find concrete evidence for a recently discovered form of intermittency, referred to as in-out intermittency, in both partial differential equation (PDE) and ordinary differential equation (ODE) models of mean field dynamos. This type of intermittency [introduced in P. Ashwin, E. Covas, and R. Tavakol, Nonlinearity 9, 563 (1999)] occurs in systems with invariant submanifolds and, as opposed to on-off intermittency which can also occur in skew product systems, it requires an absence of skew product structure. By this we mean that the dynamics on the attractor intermittent to the invariant manifold cannot be expressed simply as the dynamics on the invariant subspace forcing the transverse dynamics; the transverse dynamics will alter that tangential to the invariant subspace when one is far enough away from the invariant manifold. Since general systems with invariant submanifolds are not likely to have skew product structure, this type of behavior may be of physical relevance in a variety of dynamical settings. The models employed here to demonstrate in-out intermittency are axisymmetric mean field dynamo models which are often used to study the observed large-scale magnetic variability in the Sun and solar-type stars. The occurrence of this type of intermittency in such models may be of interest in understanding some aspects of such variabilities. (c) 2001 American Institute of Physics. PMID- 12779476 TI - Chaotic transition in a three-coupled phase-locked loop system. AB - The chaotic transition is observed in a three-coupled phase-locked loop (PLL) system in both experiments and numerical simulations. In this system, three PLL oscillators are connected with the periodic boundary condition. Intermittency is found in partially synchronized phase, in which two of three oscillators synchronize with each other and form a pair, and the chaotic transition occurs due to the recombination of synchronized pairs so that different pair is re formed. In this phase, on-off intermittency is also observed and statistical analyses are carried out for on-off intermittent time series. This intermittency is considered as a hybrid type of intermittency with both on-off intermittency and intermittency due to the recombination of synchronized pairs present in the same time series. We also show the chaotic transition phenomena in a three coupled logistic map system. (c) 2001 American Institute of Physics. PMID- 12779477 TI - Synchronization phenomena in nephron-nephron interaction. AB - Experimental data for tubular pressure oscillations in rat kidneys are analyzed in order to examine the different types of synchronization that can arise between neighboring functional units. For rats with normal blood pressure, the individual unit (the nephron) typically exhibits regular oscillations in its tubular pressure and flow variations. For such rats, both in-phase and antiphase synchronization can be demonstrated in the experimental data. For spontaneously hypertensive rats, where the pressure variations in the individual nephrons are highly irregular, signs of chaotic phase and frequency synchronization can be observed. Accounting for a hemodynamic as well as for a vascular coupling between nephrons that share a common interlobular artery, we develop a mathematical model of the pressure and flow regulation in a pair of adjacent nephrons. We show that this model, for appropriate values of the parameters, can reproduce the different types of experimentally observed synchronization. (c) 2001 American Institute of Physics. PMID- 12779478 TI - Comment on "Finding finite-time invariant manifolds in two-dimensional velocity fields" [Chaos 10, 99 (2000)]. AB - This note serves as a commentary of the paper of Haller [Chaos 10, 99 (2000)] on techniques for detecting invariant manifolds. Here we show that the criterion of Haller can be improved in two ways. First, by using the strain basis reference frame, a more efficient version of theorem 1 of Haller (2000) allows to better detect the manifolds. Second, we emphasize the need to nondimensionalize the estimate of hyperbolic persistence. These statements are illustrated by the example of the Kida ellipse. (c) 2001 American Institute of Physics. PMID- 12779479 TI - Response to "Comment on 'Finding finite-time invariant manifolds in two dimensional velocity fields' " [Chaos 11, 427 (2001)]. AB - Lapeyre, Hua, and Legras have recently suggested that the detection of finite time invariant manifolds in two-dimensional fluid flows, as described by Haller and Haller and Yuan, can be substantially improved. In particular, they suggested (a) a change of coordinates to strain basis before the application of Theorem 1 of Haller and (b) the use of a nondimensionalized time computed from Theorem 1. Here we discuss why these proposed steps will not result in a significant overall improvement. We verify our arguments in a more detailed computation of the example analyzed in Lapeyre, Hau, and Legras (the Kida ellipse), as well as in a two-dimensional barotropic turbulence simulation. While in both of these examples the techniques suggested by Lapeyre, Hau, and Legras reveal additional thin regions of hyperbolicity near vortex cores, they also lead to an overall loss of detail in the global computation of finite-time invariant manifolds. (c) 2001 American Institute of Physics. PMID- 12779480 TI - Manipulating the scaling factor of projective synchronization in three dimensional chaotic systems. AB - Scaling factor characterizes the synchronized dynamics of projective synchronization in partially linear chaotic systems but it is difficult to be estimated. To manipulate projective synchronization of chaotic systems in a favored way, a control algorithm is introduced to direct the scaling factor onto a desired value. The control approach is derived from the Lyapunov stability theory. It allows us to arbitrarily amplify or reduce the scale of the response of the slave system via a feedback control on the master system. In numerical experiments, we illustrate the application to the Lorenz system. (c) 2001 American Institute of Physics. PMID- 12779481 TI - Some topological invariants for three-dimensional flows. AB - We deal here with vector fields on three manifolds. For a system with a homoclinic orbit to a saddle-focus point, we show that the imaginary part of the complex eigenvalues is a conjugacy invariant. We show also that the ratio of the real part of the complex eigenvalue over the real one is invariant under topological equivalence. For a system with two saddle-focus points and an orbit connecting the one-dimensional invariant manifold of those points, we compute a conjugacy invariant related to the eigenvalues of the vector field at the singularities. (c) 2001 American Institute of Physics. PMID- 12779482 TI - Stability of the ground state of a harmonic oscillator in a monochromatic wave. AB - The stability of the ground state of a harmonic oscillator in a monochromatic wave is studied. This model describes, in particular, the dynamics of a cold ion in a linear ion trap, interacting with two laser fields with close frequencies. The stability of the "classical ground state"-the vicinity of the point (x=0,p=0) is analyzed analytically and numerically. For the quantum case, a method for studying a stability of the quantum ground state is developed, based on the quasienergy representation. It is demonstrated that stability of the ground state may be substantially improved by increasing the resonance number, l, where l=Omega/omega+delta, Omega and omega are, respectively, the wave frequency and the oscillator frequency, l=1,2, em leader, mid R:deltamid R:<1; or by detuning the system from exact resonance, so that delta not equal 0. The influence of a large-amplitude wave (in the presence of chaos) on the stability of the ground state is analyzed for different parameters of the model in both the quantum and classical cases. (c) 2001 American Institute of Physics. PMID- 12779483 TI - Comparison between phase space structures in coupled Morse systems and in various su(2) approximations. AB - While Hamiltonians written in terms of position and momentum provide a transparent picture of the motion of a system, Hamiltonians written in terms of Lie algebras are easier to handle quantum mechanically. Therefore we are interested to know how to transform one into the other. Since the exact transformation often leads to complicated expressions, we look for approximations which preserve the essential features. As basic criterion we look for the degree of equality of the classical phase space structures. We illustrate our ideas for the case of two coupled Morse systems and its approximation in terms of the Lie algebra su(2), which is relevant to anharmonic models of molecular spectroscopy. (c) 2001 American Institute of Physics. PMID- 12779484 TI - Robust chaos in a model of the electroencephalogram: Implications for brain dynamics. AB - Various techniques designed to extract nonlinear characteristics from experimental time series have provided no clear evidence as to whether the electroencephalogram (EEG) is chaotic. Compounding the lack of firm experimental evidence is the paucity of physiologically plausible theories of EEG that are capable of supporting nonlinear and chaotic dynamics. Here we provide evidence for the existence of chaotic dynamics in a neurophysiologically plausible continuum theory of electrocortical activity and show that the set of parameter values supporting chaos within parameter space has positive measure and exhibits fat fractal scaling. (c) 2001 American Institute of Physics. PMID- 12779485 TI - Wave propagation in distributed media. AB - The problem of chemical reaction-diffusion wave propagation through a random, heterogeneous medium is considered using a model based on cubic autocatalysis with decay. The autocatalyst is taken to diffuse and react through a reactant loaded at constant initial concentration in a reaction domain except that there may be gaps of arbitrary width in which the reactant concentration is zero. We first study the propagation of a permanent-form wave across a single gap and determine the critical width of the gap in terms of the kinetic parameters in the system. The numerical results are compared with an analytical estimate. Next, the critical conditions for propagation across two gaps separated by a domain are determined numerically, and this is extended to a series of three gaps. From these results, a series of "rules" is established to allow us to predict whether a wave will pass through an arbitrary random array of gaps of a given size subject to some imposed total void fraction for the material. (c) 2001 American Institute of Physics. PMID- 12779486 TI - Spiral wave drift induced by stimulating wave trains. AB - We investigate the drift of a spiral wave core in a homogeneous excitable medium under the influence of a periodic stimulation by wave trains close to the core. Two important results were found. First, as opposed to existing theories of spiral wave drift, we observe drift induced by wave trains with periods larger than the period of the freely rotating spiral wave. Second, when investigating the drift of meandering spirals we found that the property of meandering of spirals is not robust against periodic stimulations. Simple phenomenological arguments are provided to explain these observations. (c) 2001 American Institute of Physics. PMID- 12779487 TI - A fractional diffusion equation for a marker in porous media. AB - To study the properties of the flow of a marker through an irregular packed bed, porosity is described by stochastic processes with exponentially decaying correlation functions. We show that the ensemble average concentration of the marker satisfies a fractional diffusion equation. (c) 2001 American Institute of Physics. PMID- 12779488 TI - Global bifurcations in a laser with injected signal: Beyond Adler's approximation. AB - We discuss the dynamics in the laser with an injected signal from a perturbative point of view showing how different aspects of the dynamics get their definitive character at different orders in the perturbation scheme. At the lowest order Adler's equation [Proc. IRE 34, 351 (1946)] is recovered. More features emerge at first order including some bifurcations sets and the global reinjection conjectured in Physica D 109, 293 (1997). The type of codimension-2 bifurcations present can only be resolved at second order. We show that of the two averaging approximations proposed [Opt. Commun. 111, 173 (1994); Quantum Semiclassic. Opt. 9, 797 (1997); Quantum Semiclassic. Opt. 8, 805 (1996)] differing in the second order terms, only one is accurate to the order required, hence, solving the apparent contradiction among these results. We also show in numerical studies how a homoclinic orbit of the Sil'nikov type, bifurcates into a homoclinic tangency of a periodic orbit of vanishing amplitude. The local vector field at the transition point contains a Hopf-saddle-node singularity, which becomes degenerate and changes type. The overall global bifurcation is of codimension-3. The parameter governing this transition is theta, the cavity detuning (with respect to the atomic frequency) of the laser. (c) 2001 American Institute of Physics. PMID- 12779489 TI - Food chain chaos due to junction-fold point. AB - Consideration is given to a basic food chain model satisfying the trophic time diversification hypothesis which translates the model into a singularly perturbed system of three time scales. It is demonstrated that in some realistic system parameter region, the model has a unimodal or logistic-like Poincare return map when the singular parameter for the fastest variable is at the limiting value 0. It is also demonstrated that the unimodal map goes through a sequence of period doubling bifurcations to chaos. The mechanism for the creation of the unimodal criticality is due to the existence of a junction-fold point [B. Deng, J. Math. Biol. 38, 21-78 (1999)]. The fact that junction-fold points are structurally stable and the limiting structures persist gives us a rigorous but dynamical explanation as to why basic food chain dynamics can be chaotic. (c) 2001 American Institute of Physics. PMID- 12779490 TI - Overview: Unsolved problems of noise and fluctuations. AB - Noise and fluctuations are at the seat of all physical phenomena. It is well known that, in linear systems, noise plays a destructive role. However, an emerging paradigm for nonlinear systems is that noise can play a constructive role-in some cases information transfer can be optimized at nonzero noise levels. Another use of noise is that its measured characteristics can tell us useful information about the system itself. Problems associated with fluctuations have been studied since 1826 and this Focus Issue brings together a collection of articles that highlight some of the emerging hot unsolved noise problems to point the way for future research. (c) 2001 American Institute of Physics. PMID- 12779491 TI - Quantum vacuum noise in physics and cosmology. AB - The concept of the vacuum in quantum field theory is a subtle one. Vacuum states have a rich and complex set of properties that produce distinctive, though usually exceedingly small, physical effects. Quantum vacuum noise is familiar in optical and electronic devices, but in this paper I wish to consider extending the discussion to systems in which gravitation, or large accelerations, are important. This leads to the prediction of vacuum friction: The quantum vacuum can act in a manner reminiscent of a viscous fluid. One result is that rapidly changing gravitational fields can create particles from the vacuum, and in turn the backreaction on the gravitational dynamics operates like a damping force. I consider such effects in early universe cosmology and the theory of quantum black holes, including the possibility that the large-scale structure of the universe might be produced by quantum vacuum noise in an early inflationary phase. I also discuss the curious phenomenon that an observer who accelerates through a quantum vacuum perceives a bath of thermal radiation closely analogous to Hawking radiation from black holes, even though an inertial observer registers no particles. The effects predicted raise very deep and unresolved issues about the nature of quantum particles, the role of the observer, and the relationship between the quantum vacuum and the concepts of information and entropy. (c) 2001 American Institute of Physics. PMID- 12779492 TI - Modeling quantum measurement probability as a classical stochastic process. AB - The time-dependent measurement probabilities for the simple two-state quantum oscillator seem to invite description as a classical two-state stochastic process. It has been shown that such a description cannot be achieved using a Markov process. Constructing a more general non-Markov process is a challenging task, requiring as it does the proper generalizations of the Markovian Chapman Kolmogorov and master equations. Here we describe those non-Markovian generalizations in some detail, and we then apply them to the two-state quantum oscillator. We devise two non-Markovian processes that correctly model the measurement statistics of the oscillator, we clarify a third modeling process that was proposed earlier by others, and we exhibit numerical simulations of all three processes. Our results illuminate some interesting though widely unappreciated points in the theory of non-Markovian stochastic processes. But since quantum theory does not tell us which one of these quite different modeling processes "really" describes the behavior of the oscillator, and also since none of these processes says anything about the dynamics of other (noncommuting) oscillator observables, we can see no justification for regarding any of these processes as being fundamentally descriptive of quantum dynamics. (c) 2001 American Institute of Physics. PMID- 12779493 TI - Spatiotemporal stochastic resonance and its consequences in neural model systems. AB - The realization of spatiotemporal stochastic resonance is studied in a two dimensional FitzHugh-Nagumo system, and in a one-dimensional system of integrate and-fire neurons. We show that spatiotemporal stochastic resonance occurs in these neural model systems, independent of the method of modeling. Moreover, the ways of realization are analogous in the two model systems. The biological implications and open questions are discussed. (c) 2001 American Institute of Physics. PMID- 12779494 TI - Additive noise in noise-induced nonequilibrium transitions. AB - We study different nonlinear systems which possess noise-induced nonequlibrium transitions and shed light on the role of additive noise in these effects. We find that the influence of additive noise can be very nontrivial: it can induce first- and second-order phase transitions, can change properties of on-off intermittency, or stabilize oscillations. For the Swift-Hohenberg coupling, that is a paradigm in the study of pattern formation, we show that additive noise can cause the formation of ordered spatial patterns in distributed systems. We show also the effect of doubly stochastic resonance, which differs from stochastic resonance, because the influence of noise is twofold: multiplicative noise and coupling induce a bistability of a system, and additive noise changes a response of this noise-induced structure to the periodic driving. Despite the close similarity, we point out several important distinctions between conventional stochastic resonance and doubly stochastic resonance. Finally, we discuss open questions and possible experimental implementations. (c) 2001 American Institute of Physics. PMID- 12779495 TI - Noise-assisted traffic of spikes through neuronal junctions. AB - The presence of noise, i.e., random fluctuations, in the nervous system raises at least two different questions. First, is there a constructive role noise can play for signal transmission in a neuron channel? Second, what is the advantage of the power spectra observed for the neuron activity to be shaped like 1/f(k)? To address these questions a simple stochastic model for a junction in neural spike traffic channels is presented. Side channel traffic enters main channel traffic depending on the spike rate of the latter one. The main channel traffic itself is triggered by various noise processes such as Poissonian noise or the zero crossings of Gaussian 1/f(k) noise whereas the variation of the exponent k gives rise to a maximum of the overall traffic efficiency. It is shown that the colored noise is superior to the Poissonian and, in certain cases, to deterministic, periodically ordered traffic. Further, if this periodicity itself is modulated by Gaussian noise with different spectral exponents k, then such modulation can lead to noise-assisted traffic as well. The model presented can also be used to consider car traffic at a junction between a main and a side road and to show how randomness can enhance the traffic efficiency in a network. (c) 2001 American Institute of Physics. PMID- 12779496 TI - Activated escape of periodically driven systems. AB - We discuss activated escape from a metastable state of a system driven by a time periodic force. We show that the escape probabilities can be changed very strongly even by a comparatively weak force. In a broad parameter range, the activation energy of escape depends linearly on the force amplitude. This dependence is described by the logarithmic susceptibility, which is analyzed theoretically and through analog and digital simulations. A closed-form explicit expression for the escape rate of an overdamped Brownian particle is presented and shown to be in quantitative agreement with the simulations. We also describe experiments on a Brownian particle optically trapped in a double-well potential. A suitable periodic modulation of the optical intensity breaks the spatio temporal symmetry of an otherwise spatially symmetric system. This has allowed us to localize a particle in one of the symmetric wells. (c) 2001 American Institute of Physics. PMID- 12779497 TI - Noise-induced escape on time scales preceding quasistationarity: New developments in the Kramers problem. AB - Noise-induced escape from the metastable part of a potential is considered on time scales preceding the formation of quasiequilibrium within that part of the potential. It is shown that, counterintuitively, the escape flux may then depend exponentially strongly, and in a complicated manner, on time and friction. (c) 2001 American Institute of Physics. PMID- 12779498 TI - "Green" noise in quasistationary stochastic systems. AB - We consider the behavior of stochastic systems driven by noise with a zero value of spectral density at zero frequency ("green" noise). For this purpose we propose the version of the Krylov-Bogoliubov averaging method to study the systems which are not stationary in the case of an external white noise. We use the ergodicity of a nonlinear random function in the method, and obtain equations for any approximation of the theory. In particular, it is shown in the first approximation that there is an effective potential to describe the averaged motion of the system. We consider a phase-locked loop as an example and show that metastable states are possible. The lifetime of these states essentially increases if the form of a green noise spectrum becomes sharper in the low frequency region. The high stability of the system driven by green noise is confirmed by numerical simulation. It is important that the theoretical result obtained by the averaging method and the one obtained in the simulation coincide with sufficient accuracy. In conclusion, we discuss some of the unsolved green noise problems. (c) 2001 American Institute of Physics. PMID- 12779499 TI - Level-crossing statistics of the horizontal wind speed in the planetary surface boundary layer. AB - The probability density of the times for which the horizontal wind remains above or below a given threshold speed is of some interest in the fields of renewable energy generation and pollutant dispersal. However there appear to be no analytic or conceptual models which account for the observed power law form of the distribution of these episode lengths over a range of over three decades, from a few tens of seconds to a day or more. We reanalyze high resolution wind data and demonstrate the fractal character of the point process generated by the wind speed level crossings. We simulate the fluctuating wind speed by a Markov process which approximates the characteristics of the real (non-Markovian) wind and successfully generates a power law distribution of episode lengths. However, fundamental questions concerning the physical basis for this behavior and the connection between the properties of a continuous-time stochastic process and the fractal statistics of the point process generated by its level crossings remain unanswered. (c) 2001 American Institute of Physics. PMID- 12779500 TI - Amplitude truncation of Gaussian 1/f(alpha) noises: Results and problems. AB - An interesting property of Gaussian 1/f noise was found experimentally a few years ago: The amplitude truncation does not change the power spectral density of the noise under rather general conditions. Here we present a brief theoretical derivation of this invariant property of band-limited Gaussian 1/f noise and include 1/f(alpha) noises also with 0 infinity. We also study these networks numerically. Using three different measures (types of attractors, fraction of elements that are active, and length of period), we show that finite, scale-free networks are more ordered than either the Poisson or delta function networks below the critical point. Thus the topology of scale-free biochemical networks, characterized by a wide distribution in the number of inputs per element, may provide a source of order in living cells. (c) 2001 American Institute of Physics. PMID- 12779521 TI - Lagrangian chaos and Eulerian chaos in shear flow dynamics. AB - Shear flow dynamics described by the two-dimensional incompressible Navier-Stokes equations is studied for a one-dimensional equilibrium vorticity profile having two minima. These lead to two linear Kelvin-Helmholtz instabilities; the resulting nonlinear waves corresponding to the two minima have different phase velocities. The nonlinear behavior is studied as a function of two parameters, the Reynolds number and a parameter lambda specifying the width of the minima in the vorticity profile. For parameters such that the instabilities grow to a sufficient level, there is Lagrangian chaos, leading to mixing of vorticity, i.e., momentum transport, between the chains of vortices or cat's eyes. Lagrangian chaos is quantified by plotting the finite time Lyapunov exponents on a grid of initial points, and by the probability distribution of these exponents. For moderate values of lambda, there is Lagrangian chaos everywhere except near the centers of the vortices and near the boundaries, and there are competing effects of homogenization of vorticity and formation of structures associated with secondary resonances. For smaller values of lambda Lagrangian chaos occurs in the regions in the centers of the vortices, and the Eulerian behavior of the flow undergoes bifurcations leading to Eulerian chaos, as measured by the time series of several Galilean invariant quantities. A discussion of Lagrangian chaos and its relation to Eulerian chaos is given.(c) 2001 American Institute of Physics. PMID- 12779522 TI - Dynamics of one- and two-dimensional kinks in bistable reaction-diffusion equations with quasidiscrete sources of reaction. AB - We study the evolution of fronts in a bistable reaction-diffusion system when the nonlinear reaction term is spatially inhomogeneous. This equation has been used to model wave propagation in various biological systems. Extending previous works on homogeneous reaction terms, we derive asymptotically an equation governing the front motion, which is strongly nonlinear and, for the two-dimensional case, generalizes the classical mean curvature flow equation. We study the motion of one- and two-dimensional fronts, finding that the inhomogeneity acts as a "potential function" for the motion of the front; i.e., there is wave propagation failure and the steady state solution depends on the structure of the function describing the inhomogeneity. (c) 2001 American Institute of Physics. PMID- 12779523 TI - A new approximate analytical approach for dispersion relation of the nonlinear Klein-Gordon equation. AB - A novel approach is presented for obtaining approximate analytical expressions for the dispersion relation of periodic wavetrains in the nonlinear Klein-Gordon equation with even potential function. By coupling linearization of the governing equation with the method of harmonic balance, we establish two general analytical approximate formulas for the dispersion relation, which depends on the amplitude of the periodic wavetrain. These formulas are valid for small as well as large amplitude of the wavetrain. They are also applicable to the large amplitude regime, which the conventional perturbation method fails to provide any solution, of the nonlinear system under study. Three examples are demonstrated to illustrate the excellent approximate solutions of the proposed formulas with respect to the exact solutions of the dispersion relation. (c) 2001 American Institute of Physics. PMID- 12779524 TI - Complexity of regular invertible p-adic motions. AB - We consider issues of computational complexity that arise in the study of quasi periodic motions (Siegel discs) over the p-adic integers, where p is a prime number. These systems generate regular invertible dynamics over the integers modulo p(k), for all k, and the main questions concern the computation of periods and orbit structure. For a specific family of polynomial maps, we identify conditions under which the cycle structure is determined solely by the number of Siegel discs and two integer parameters for each disc. We conjecture the minimal parametrization needed to achieve-for every odd prime p-a two-disc tessellation with maximal cycle length. We discuss the relevance of Cebotarev's density theorem to the probabilistic description of these dynamical systems. (c) 2001 American Institute of Physics. PMID- 12779525 TI - On the wavelet formalism for multifractal analysis. AB - It is proved that the multifractal characterizations of diametrically regular measures that are provided by the wavelet and by the Hentschel-Procaccia formalisms are identical. (c) 2001 American Institute of Physics. PMID- 12779526 TI - Control of long-period orbits and arbitrary trajectories in chaotic systems using dynamic limiting. AB - We demonstrate experimental control of long-period orbits and arbitrary chaotic trajectories using a new chaos control technique called dynamic limiting. Based on limiter control, dynamic limiting uses a predetermined sequence of limiter levels applied to the chaotic system to stabilize natural states of the system. The limiter sequence is clocked by the natural return time of the chaotic system such that the oscillator sees a new limiter level for each peak return. We demonstrate control of period-8 and period-34 unstable periodic orbits in a low frequency circuit and provide evidence that the control perturbations are minimal. We also demonstrate control of an arbitrary waveform by replaying a sequence captured from the uncontrolled oscillator, achieving a form of delayed self-synchronization. Finally, we discuss the use of dynamic limiting for high frequency chaos communications. (c) 2002 American Institute of Physics. PMID- 12779527 TI - Dispersion management for solitons in a Korteweg-de Vries system. AB - The existence of "dispersion-managed solitons," i.e., stable pulsating solitary wave solutions to the nonlinear Schrodinger equation with periodically modulated and sign-variable dispersion is now well known in nonlinear optics. Our purpose here is to investigate whether similar structures exist for other well-known nonlinear wave models. Hence, here we consider as a basic model the variable coefficient Korteweg-de Vries equation; this has the form of a Korteweg-de Vries equation with a periodically varying third-order dispersion coefficient, that can take both positive and negative values. More generally, this model may be extended to include fifth-order dispersion. Such models may describe, for instance, periodically modulated waveguides for long gravity-capillary waves. We develop an analytical approximation for solitary waves in the weakly nonlinear case, from which it is possible to obtain a reduction to a relatively simple integral equation, which is readily solved numerically. Then, we describe some systematic direct simulations of the full equation, which use the soliton shape produced by the integral equation as an initial condition. These simulations reveal regions of stable and unstable pulsating solitary waves in the corresponding parametric space. Finally, we consider the effects of fifth-order dispersion. (c) 2002 American Institute of Physics. PMID- 12779528 TI - Tracking target and spiral waves. AB - A new algorithm for analyzing the evolution of patterns of spiral and target waves in large aspect ratio chemical systems is introduced. The algorithm does not depend on finding the spiral tip but locates the center of the pattern by a new concept, called the spiral focus, which is defined by the evolutes of the actual spiral or target wave. With the use of Gaussian smoothing, a robust method is developed that permits the identification of targets and spirals foci independently of the wave profile. Examples of an analysis of long image sequences from experiments with the Belousov-Zhabotinsky reaction catalyzed by ruthenium-tris-bipyridyl are presented. Moving target and spiral foci are found, and the speed and direction of movement of single as well as double spiral foci are investigated. For the experiments analyzed in this paper it is found that the movement of a focus correlates with foci in the immediate neighborhood independently of how they were created. (c) 2002 American Institute of Physics. PMID- 12779529 TI - An application of the least-squares method to system parameters extraction from experimental data. AB - We explore the possibility and the limits of extracting the parameters of the model from simulated logistic and Henon time series. For the models considered, the least-squares approach provides accurate values of the recurrence order and polynomial degree along with the model parameters. We found that the number of data points increases the accuracy of the estimation only for noise-free data. With the white noise added to the data, the accuracy could not be improved above a certain threshold that is almost independent of the number of data points. The additive noise flattened the global minimum of the least-squares function such that above a noise threshold it is no longer possible to discern the optimum values of the recurrence order and/or polynomial degree. (c) 2002 American Institute of Physics. PMID- 12779530 TI - Dynamical ordering of symmetric non-Birkhoff periodic points in reversible monotone twist mappings. AB - We study the coexistence of symmetric non-Birkhoff periodic orbits of C(1) reversible monotone twist mappings on the cylinder. We prove the equivalence of the existence of non-Birkhoff periodic orbits and that of transverse homoclinic intersections of stable and unstable manifolds of the fixed point. We derive the positional relation of symmetric Birkhoff and non-Birkhoff periodic orbits and obtain the dynamical ordering of symmetric non-Birkhoff periodic orbits. An extension of the Sharkovskii ordering to two-dimensional mappings has been carried out. In the proof of various properties of the mappings, reversibility plays an essential role. (c) 2002 American Institute of Physics. PMID- 12779531 TI - Chaotic maps derived from trajectory data. AB - We assume that we have the following information from observed trajectory data about a discrete time dynamical process: (1) a rough graph of the phase plot, (2) the probability density function and (3) the metric entropy. We describe a method for constructing a nonlinear map whose graph is "within" the rough graph of the phase plot, and which has the observed probability density function and entropy.(c) 2002 American Institute of Physics. PMID- 12779532 TI - Convection in chemical fronts with quadratic and cubic autocatalysis. AB - Convection in chemical fronts enhances the speed and determines the curvature of the front. Convection is due to density gradients across the front. Fronts propagating in narrow vertical tubes do not exhibit convection, while convection develops in tubes of larger diameter. The transition to convection is determined not only by the tube diameter, but also by the type of chemical reaction. We determine the transition to convection for chemical fronts with quadratic and cubic autocatalysis. We show that quadratic fronts are more stable to convection than cubic fronts. We compare these results to a thin front approximation based on an eikonal relation. In contrast to the thin front approximation, reaction diffusion models show a transition to convection that depends on the ratio between the kinematic viscosity and the molecular diffusivity. (c) 2002 American Institute of Physics. PMID- 12779533 TI - The effect of reactor geometry on frontal polymerization spin modes. AB - Using reactors of different sizes and geometries the dynamics of the frontal polymerization of 1,6-hexanediol diacrylate (HDDA) and pentaerythritol tetraacrylate (PETAC), with ammonium persulfate as the initiator were studied. For this system, the frontal polymerization exhibits complex behavior that depends on the ratio of the monomers. For a particular range of monomers concentration, the polymerization front becomes nonplanar, and spin modes appear. By varying the reactor diameter, we experimentally confirmed the expected shift of the system to a greater number of "hot spots" for larger diameters. For square test tubes a "zig-zag" mode was observed for the first time in frontal polymerization. We confirmed the viscosity-dependence of the spin mode instabilities. We also observed novel modes in cylinder-inside-cylinder reactors. Lastly, using a conical reactor with a continuously varying diameter, we observed what may be evidence for bistability depending on the direction of propagation. We discuss these finding in terms of the standard linear stability analysis for propagating fronts. (c) 2002 American Institute of Physics. PMID- 12779534 TI - A note on discretization of nonlinear differential equations. AB - An important issue when integrating nonlinear differential equations on a digital computer is the choice of the time increment or step size. For example, it is known that if this quantity is not sufficiently short, spurious chaotic motions may be induced when integrating a system using several of the well-known methods available in the literature. In this paper, a new approach to discretize differential equations is analyzed in light of computational chaos. It will be shown that the fixed points of the continuous system are preserved under the new discretization approach and that the spurious fixed points generated by higher order approximations depend upon the increment parameter. (c) 2002 American Institute of Physics. PMID- 12779535 TI - Symmetry breaking, bifurcations, quasiperiodicity, and chaos due to electric fields in a coupled cell model. AB - A model for the asymmetric coupling of two oscillatory cells is considered. The coupling between the cells is both through diffusional exchange (symmetric) and through the electromigration of ionic reactant species from one cell to the other (asymmetric) in applied electric fields. The kinetics in each cell are the same and based on the Gray-Scott scheme. Without the electric field, only simple, stable dynamics are seen. The effect of the asymmetry (applying electric fields) is to create a wide variety of stable dynamics, multistability, multiperiodic oscillations, quasiperiodicity and chaos being observed, this complexity in response being more prevalent at weaker coupling rates and at weaker field strengths. The results are obtained using a standard dynamical systems continuation program, though asymptotic results are obtained for strong coupling rates and strong electric fields. These are seen to agree well with the numerically determined values in the appropriate parameter regimes. (c) 2002 American Institute of Physics. PMID- 12779536 TI - Perturbed ion traps: A generalization of the three-dimensional Henon-Heiles problem. AB - This paper presents an analytical study of an axially symmetric perturbation of the Penning trap. This system is modeled as a generalization of the three dimensional (3D) Henon-Heiles potential. Thus, the same techniques which succeeded in the study of the 3D Henon-Heiles system apply here. The departure Hamiltonian is three dimensional, although it possesses an axial symmetry. This property, together with an averaging process, is used to reduce the original system to an integrable one. We study the flow of the reduced Hamiltonian: equilibria, bifurcations, and stability, extracting thereafter the relevant information about the dynamics of the original problem. (c) 2002 American Institute of Physics. PMID- 12779537 TI - n:m phase synchronization with mutual coupling phase signals. AB - We generalize the n:m phase synchronization between two chaotic oscillators by mutual coupling phase signals. To characterize this phenomenon, we use two coupled oscillators to demonstrate their phase synchronization with amplitudes practically noncorrelated. We take the 1:1 phase synchronization as an example to show the properties of mean frequencies, mean phase difference, and Lyapunov exponents at various values of coupling strength. The phase difference increases with 2pi phase slips below the transition. The scaling rules of the slip near and away from the transition are studied. Furthermore, we demonstrate the transition to a variety of n:m phase synchronizations and analyze the corresponding coupling dynamics. (c) 2002 American Institute of Physics. PMID- 12779538 TI - Introduction: Nonlinear pattern formation in surface science. AB - (c) 2002 American Institute of Physics. PMID- 12779539 TI - Microscopic aspects of pattern formation on surfaces. AB - Recent scanning tunneling microscopy (STM) work gives insight into microscopic processes of surface reactions that play a role for spatio-temporal pattern formation. STM allows to resolve adsorbed particles, follow their surface motion, and monitor reactions with other particles on the atomic scale. The data reveal pronounced deviations from the implicite assumptions of the reaction-diffusion equations traditionally used to model spatio-temporal patterns. In contrast to these descriptions, particles are often not randomly distributed, but cluster in islands because of attractive interactions, and particle hopping can be highly correlated. It is shown that such phenomena can even affect the macroscopic kinetics. The article also discusses a case where the atomic processes inside propagating reaction fronts could be resolved. Here particular strong interaction effects were observed, caused by hydrogen bonds between the reacting species. (c) 2002 American Institute of Physics. PMID- 12779540 TI - Kinetic instabilities during the NO(x) reduction with hydrogen on Pt crystals studied with field emission on the nanoscale. AB - This paper reviews field emission studies of kinetic instabilities occurring during the catalytic reduction of nitric oxide (NO) and nitrogen dioxide (NO(2)) by hydrogen on three-dimensional platinum crystals. Emphasis is placed on revealing that both field ion microscopy (FIM) and field electron microscopy (FEM) can image such instabilities under truly in situ reaction conditions with a lateral resolution on the nanoscale. In particular, oscillatory behavior with rapid ignition from a state of low to a state of high catalytic activity is demonstrated for both NO and NO(2) reduction. Results of a local chemical probing during FIM studies of the NO+H(2) reaction are also shown and provide clear evidence for the oscillatory behavior of water (detected as H(2)O(+) and H(3)O(+)) formation and for diffusion supply of NO into surface regions emptied during the stage of high catalytic activity. The rapid ignition ("surface explosion") of the catalytic cycle is discussed on the basis of an autocatalytic mechanism of the NO decomposition. On the (001) plane of the Pt crystal small island formation is seen to occur during the low-activity state of the catalytic cycle. Islands have a size equivalent to approximately 3 nm, move independently from each other, and do not merge when colliding. A tentative model is discussed associating islands with patches of hydroxyl groups. Very regular oscillatory behavior is demonstrated for the NO(2) reduction using FEM. Advantages as well as shortcomings of the FEM/FIM experimental approach are discussed and an outlook on future studies using local chemical probing will be given wherever appropriate. (c) 2002 American Institute of Physics. PMID- 12779541 TI - From atomistic lattice-gas models for surface reactions to hydrodynamic reaction diffusion equations. AB - Atomistic lattice-gas models for surface reactions can accurately describe spatial correlations and ordering in chemisorbed layers due to adspecies interactions or due to limited mobility of some adspecies. The primary challenge in such modeling is to describe spatiotemporal behavior in the physically relevant "hydrodynamic" regime of rapid diffusion of (at least some) reactant adspecies. For such models, we discuss the development of exact reaction diffusion equations (RDEs) describing mesoscale spatial pattern formation in surface reactions. Formulation and implementation of these RDEs requires detailed analysis of chemical diffusion in mixed reactant adlayers, as well as development of novel hybrid and parallel simulation techniques. (c) 2002 American Institute of Physics. PMID- 12779542 TI - Self-organized nanostructures in surface chemical reactions: Mechanisms and mesoscopic modeling. AB - Nanoscale patterns can form in reactive adsorbates on catalytic surfaces as a result of attractive lateral interactions. These structures can be described within a mesoscopic theory that is derived by coarse graining the microscopic master equation thus providing a link between microscopic lattice models and reaction-diffusion equations. Such mesoscopic models allow to systematically investigate mechanisms responsible for the formation of nanoscale nonequilibrium patterns in reactive condensed matter. We have found that stationary and traveling nanostructures may result from the interplay of the attractive lateral interactions and nonequilibrium reactions. Besides reviewing these results, a detailed investigation of a single reactive adsorbate in the presence of attractive lateral interactions and global coupling through the gas phase is presented. Finally, it is outlined how a mesoscopic theory should be constructed for a particular scanning tunneling microscopy experiment [the oxidation of hydrogen on a Pt(111) surface] in order to overcome the failure of a corresponding reaction-diffusion model to quantitatively reproduce the experiments. (c) 2002 American Institute of Physics. PMID- 12779543 TI - Pattern formation during the CO-oxidation involving subsurface oxygen. AB - This paper focuses on subsurface oxygen and its influence on pattern formation during CO-oxidation on platinum surfaces. For the observation of spatiotemporal pattern formation during catalytic reactions the photoelectron emission microscope (PEEM) has proven to be an excellent real-time imaging instrument, capable of tracking local work function changes. The existence of subsurface oxygen on platinumlike surfaces has been extensively discussed and for palladium its presence has been clearly established during rate oscillations. Subsurface oxygen is defined at this point as an atomic O species located directly underneath the uppermost metal crystal layer; its dipole moment therefore considerably lowers the work function of the surface. Here we review some of the investigations involving subsurface oxygen, focusing on the role subsurface oxygen might play in pattern formation during CO-oxidation on platinum. We will also present some new results, where this species clearly interacts with chemisorbed oxygen under restrictions by boundary conditions on the Pt(110) single crystal. These previously (through microlithography) constructed domain boundaries on the surface are made out of Rh or Pd, and they are acting as an additional source of CO molecules for the Pt surface. (c) 2002 American Institute of Physics. PMID- 12779544 TI - Spatio-temporal pattern formation during CO oxidation on Pt(100) at low and intermediate pressures: A comparative study. AB - Experimental studies of CO oxidation on Pt(100) over two different ranges of reactant pressures will be reviewed. Using photoemission electron microscopy (PEEM), spatio-temporal pattern formation was observed at temperatures between 420 and 540 K in the 10(-5) mbar pressure range. In an attempt to bridge the "pressure-gap," ellipsomicroscopy for surface imaging was used to follow pattern formation at temperatures around 600 K in the 10(-2) mbar pressure range. The features of the nonlinear phenomena, observed in these two different pressure regimes, are markedly different. This is shown by comparison of various qualitative and quantitative features of spatio-temporal pattern formation as well as the dynamics of the macroscopic reaction rate. Subsurface oxygen is proposed as a tentative alternative to the surface phase transition for oscillations in the reaction rate at higher temperatures and intermediate pressures. (c) 2002 American Institute of Physics. PMID- 12779545 TI - Hot zones evolution and dynamics in heterogeneous catalytic systems. AB - Stationary and complex moving hot regions formed for temperatures close to the extinction temperature of uniformly ignited states of several catalytic systems, such as thin rings and hollow cylinders, a thin radial flow reactor (RFR) and a shallow packed bed. IR imaging revealed that the hot and cold regions (temperature difference of the order of 100 degrees C) were separated by a sharp (about 3 mm wide) temperature front. The transition from the branch of uniformly ignited to the states with a hot region was usually supercritical. In some experiments a disjoint branch of states with hot regions existed and two qualitatively different states with hot zones existed under the same operating conditions. A very intricate periodic motion of a hot zone was observed in a shallow packed bed reactor. For example, Fig. 16 shows a hot zone which splits and later coalesces several times during the long (14 h) period. Hot pulse motions were observed on a single catalytic pellet. These were caused by global coupling between the surface reaction rate and the ambient reactant concentration and the inherent nonuniformity of the catalytic activity. It is not yet clear what rate processes generate the transversal hot zones in uniform packed bed reactors. (c) 2002 American Institute of Physics. PMID- 12779546 TI - Catalysis on microstructured bimetallic surfaces. AB - Microstructured bimetallic Pt/Rh and Pt/Ti surfaces have been employed to study the dynamics of catalytic NO reduction and the O(2)+H(2) reaction at low pressure (p<10(-3) mbar). Photoelectron emission microscopy and scanning photoelectron microscopy were used as spatially resolved in situ methods to image the local work function changes and to identify chemical changes in the substrate and in the adsorbate layer. It is shown that diffusional coupling leads to dynamic effects which are dependent on the macroscopic size (&mgr;m range). With alkali metals on the surface, stationary patterns form whose mechanism of formation has been studied in detail. (c) 2002 American Institute of Physics. PMID- 12779547 TI - Front initiation on microdesigned composite catalysts. AB - We first briefly review the subject of spatiotemporal pattern formation on microdesigned composite catalysts. One of the most significant interaction mechanisms between different reacting domains (consisting of different metal catalysts such as Pt and Rh, coupled through surface diffusion) is the initiation of reaction fronts at the interface between them. We then explore in some detail the effect of two-dimensional composite geometry on this basic building block of composite catalyst dynamics. (c) 2002 American Institute of Physics. PMID- 12779548 TI - Pattern formation on anisotropic and heterogeneous catalytic surfaces. AB - We review experimental and theoretical work addressing pattern formation on anisotropic and heterogeneous catalytic surfaces. These systems are typically modeled by reaction-diffusion equations reflecting the kinetics and transport of the involved chemical species. Here, we demonstrate the influence of anisotropy and heterogeneity in a simplified model, the FitzHugh-Nagumo equations. Anisotropy causes stratification of labyrinthine patterns and spiral defect chaos in bistable media. For heterogeneous media, we study the situation where the heterogeneity appears on a length scale shorter than the typical pattern length scale. Homogenization, i.e., computation of effective medium properties, is applied to an example and illustrated with simulations in one (fronts) and two dimensions (spirals). We conclude with a discussion of open questions and promising directions that comprise the coupling of the microscopic structure of the surface to the macroscopic concentration patterns and the fabrication of nanostructures with heterogeneous surfaces as templates. (c) 2002 American Institute of Physics. PMID- 12779549 TI - Theory of electrochemical pattern formation. AB - The spatial coupling in electrochemical systems is mediated by ion migration under the influence of the electric field. Since field effects spread very rapidly, every point of an electrode can communicate with every other one practically instantaneously through migration coupling. Based on mathematical potential theory we present the derivation of a generally applicable reaction migration equation, which describes the coupling via an integral over the whole electrode area. The corresponding coupling function depends only on the geometry of the electrode setup and has been computed for commonly used electrode shapes (such as ring, disk, ribbon or rectangle). The pattern formation observed in electrochemical systems in the bistable, excitable and oscillatory regime can be reproduced in computer simulations, and the types of patterns occurring under different geometries can be rationalized. (c) 2002 American Institute of Physics. PMID- 12779550 TI - Evolution of spatiotemporal patterns during the electrodissolution of metals: Experiments and simulations. AB - Spatiotemporal patterns including accelerating fronts, rotating waves, and homogeneous oscillations evolve during the electrodissolution of metals like cobalt and iron that exhibit passivity under potentiostatic control. The nature of the patterns is determined by long-range (nonlocal) coupling through the electric field which in turn is influenced by the geometry of the electrochemical cell, the applied potential, and the conductivity of the electrolyte. A two variable model in a three-dimensional geometry is presented which is able to simulate the essential features of the experimental system.(c) 2002 American Institute of Physics. PMID- 12779552 TI - Populations of coupled electrochemical oscillators. AB - Experiments were carried out on arrays of chaotic electrochemical oscillators to which global coupling, periodic forcing, and feedback were applied. The global coupling converts a very weakly coupled set of chaotic oscillators to a synchronized state with sufficiently large values of coupling strength; at intermediate values both intermittent and stable chaotic cluster states occur. Cluster formation and synchronization were also obtained by applying feedback and forcing to a moderately coupled base state. The three cases differ, however, in other details. The feedback and forcing also produce periodic cluster states and more than two clusters. Configurations of two (chaotic) clusters and two, three, or four (periodic) clusters were observed. (c) 2002 American Institute of Physics. PMID- 12779551 TI - Nanoscale pore formation dynamics during aluminum anodization. AB - A theoretical analysis of nanoscale pore formation during anodization reveals its fundamental instability mechanism to be a field focusing phenomenon when perturbations on the minima of the two oxide interfaces are in phase. Lateral leakage of the layer potential at high wave number introduces a layer tension effect that balances the previous destabilizing effect to produce a long-wave instability and a selected pore separation that scales linearly with respect to voltage. At pH higher than 1.77, pores do not form due to a very thick barrier layer. A weakly nonlinear theory based on long-wave expansion of double free surface problem yields two coupled interface evolution equations that can be reduced to one without altering the dispersion relationship by assuming an equal and in-phase amplitude for the two interfaces. This interface evolution equation faithfully reproduces the initial pore ordering and their dynamics. A hodograph transformation technique is then used to determine the interior dimension of the well-developed pores in two dimensions. The ratio of pore diameter to pore separation is found to be a factor independent of voltage but varies with the pH of the electrolyte. Both the predicted pH range where pores are formed and the predicted pore dimensions are favorably compared to experimental data. (c) 2002 American Institute of Physics. PMID- 12779553 TI - Erratum: "A fractional diffusion equation for a marker in porous media" [Chaos 11, 495 (2001)]. PMID- 12779554 TI - A three-dimensional renormalization group bubble merger model for Rayleigh-Taylor mixing. AB - In this paper we formulate a model for the merger of bubbles at the edge of an unstable acceleration driven (Rayleigh-Taylor) mixing layer. Steady acceleration defines a self-similar mixing process, with a time-dependent inverse cascade of structures of increasing size. The time evolution is itself a renormalization group (RNG) evolution, and so the large time asymptotics define a RNG fixed point. We solve the model introduced here at this fixed point. The model predicts the growth rate of a Rayleigh-Taylor chaotic fluid mixing layer. The model has three main components: the velocity of a single bubble in this unstable flow regime, an envelope velocity, which describes collective excitations in the mixing region, and a merger process, which drives an inverse cascade, with a steady increase of bubble size. The present model differs from an earlier two dimensional (2-D) merger model in several important ways. Beyond the extension of the model to three dimensions, the present model contains one phenomenological parameter, the variance of the bubble radii at fixed time. The model also predicts several experimental numbers: the bubble mixing rate, alpha(b)=h(b)/Agt(2) approximately 0.05-0.06, the mean bubble radius, and the bubble height separation at the time of merger. From these we also obtain the bubble height to the radius aspect ratio. Using the experimental results of Smeeton and Youngs (AWE Report No. O 35/87, 1987) to fix a value for the radius variance, we determine alpha(b) within the range of experimental uncertainty. We also obtain the experimental values for the bubble height to width aspect ratio in agreement with experimental values. (c) 2002 American Institute of Physics. PMID- 12779555 TI - Noise-resistant chaotic maps. AB - Synchronized chaotic systems are highly vulnerable to noise added to the synchronizing signal. It was previously shown that chaotic circuits could be built that were less sensitive to this type of noise. In this work, simple chaotic maps are demonstrated that are also less sensitive to added noise. These maps are based on coupling a shift map to a digital filter. These maps are simple enough that they should help lead to an understanding of how noise-robust chaotic systems work. (c) 2002 American Institute of Physics. PMID- 12779556 TI - A class of cellular automata modeling winnerless competition. AB - Neural units introduced by Rabinovich et al. ("Sensory coding with dynamically competitive networks," UCSD and CIT, February 1999) motivate a class of cellular automata (CA) where spatio-temporal encoding is feasible. The spatio-temporal information capacity of a CA is estimated by the information capacity of the attractor set, which happens to be finitely specified. Two-dimensional CA are studied in detail. An example is given for which the attractor is not a subshift. (c) 2002 American Institute of Physics. PMID- 12779557 TI - Volume-preserving maps with an invariant. AB - Several families of volume-preserving maps on R(3) that have an integral are constructed using techniques due to Suris. We study the dynamics of these maps as the topology of the two-dimensional level sets of the invariant changes. (c) 2002 American Institute of Physics. PMID- 12779558 TI - Successive bifurcations in a simple model of atmospheric zonal-flow vacillation. AB - Low-frequency variability of the atmospheric flow in the Southern Hemisphere is dominated by irregular changes in the latitude and intensity of the mid-latitude eastward jet about its climatological mean state. This phenomenon, known as atmospheric zonal-flow vacillation, is characterized by the existence of two persistent states of the zonal (i.e., east-west oriented) jet and irregular transitions between them. Nonlinear interactions between the mean flow and the waves play a key role in the dynamics of this vacillation. In the present study, we develop a low-order, deterministic model for the nonlinear dynamics of atmospheric zonal-flow vacillation. Multiple equilibria arise in this model's zonal-mean flow, that is, in the longitudinal flow averaged along a given latitude circle. These equilibria bear a strong resemblance to the two persistent flow regimes found in Southern Hemisphere observations. The two equilibrium states are maintained by wave forcing against surface drag, as in the observations. Successive bifurcations to periodic and chaotic zonal-mean flow regimes occur as the model's dissipation parameter is reduced. (c) 2002 American Institute of Physics. PMID- 12779559 TI - Length of magnetic field lines in turbulent plasmas. AB - An estimation of the length of any magnetic field line in a two-dimensional periodic magnetohydrodynamic problem is provided. This is done by using some classical function theory results on the analytic extension of the vector potential. The essential parameter, the maximum of this extension, may be analyzed in the case of turbulent plasmas by admitting the Iroshnikov-Kraichnan statistics, establishing in this way a relation between the length of any magnetic field line and the energy dissipation scale. (c) 2002 American Institute of Physics. PMID- 12779560 TI - On the geometry of master-slave synchronization. AB - In 1990, Pecora and Carroll reported the observation that one can synchronize the orbits of two identical dynamical systems, which may be chaotic, by feeding state variables of one of them to the other one with no feedback, a phenomenon often called master-slave synchronization. We report here some results on the theory of master-slave synchronization for maps and flows, which are all inspired by a similar geometric and coordinate independent point of view to the one introduced in master-slave synchronization by Tresser, Worfolk, and Bass. Our results are variations on the theme that projection often can compensate for expansion.(c) 2002 American Institute of Physics. PMID- 12779561 TI - Short-lived two-soliton bound states in weakly perturbed nonlinear Schrodinger equation. AB - Resonant soliton collisions in the weakly discrete nonlinear Schrodinger equation are studied numerically. The fractal nature of the soliton scattering, described in our previous works, is investigated in detail. We demonstrate that the fractal scattering pattern is related to the existence of the short-lived two-soliton bound states. The bound state can be regarded as a two-soliton quasiparticle of a new type, different from the breather. We establish that the probability P of a bound state with the lifetime L follows the law P approximately L(-3). In the frame of a simple two-particle model, we derive the nonlinear map, which generates the fractal pattern similar to that observed in the numerical study of soliton collisions. (c) 2002 American Institute of Physics. PMID- 12779562 TI - Complex dynamics in a simple model of pulsations for super-asymptotic giant branch stars. AB - When intermediate mass stars reach their last stages of evolution they show pronounced oscillations. This phenomenon happens when these stars reach the so called asymptotic giant branch (AGB), which is a region of the Hertzsprung Russell diagram located at about the same region of effective temperatures but at larger luminosities than those of regular giant stars. The period of these oscillations depends on the mass of the star. There is growing evidence that these oscillations are highly correlated with mass loss and that, as the mass loss increases, the pulsations become more chaotic. In this paper we study a simple oscillator which accounts for the observed properties of this kind of stars. This oscillator was first proposed and studied in Icke et al. [Astron. Astrophys. 258, 341 (1992)] and we extend their study to the region of more massive and luminous stars -the region of super-AGB stars. The oscillator consists of a periodic nonlinear perturbation of a linear Hamiltonian system. The formalism of dynamical systems theory has been used to explore the associated Poincare map for the range of parameters typical of those stars. We have studied and characterized the dynamical behavior of the oscillator as the parameters of the model are varied, leading us to explore a sequence of local and global bifurcations. Among these, a tripling bifurcation is remarkable, which allows us to show that the Poincare map is a nontwist area preserving map. Meandering curves, hierarchical-islands traps and sticky orbits also show up. We discuss the implications of the stickiness phenomenon in the evolution and stability of the super-AGB stars. (c) 2002 American Institute of Physics. PMID- 12779563 TI - Generating chaos with a switching piecewise-linear controller. AB - This paper introduces a new chaos generator, a switching piecewise-linear controller, which can create chaos from a three-dimensional linear system within a wide range of parameter values. Basic dynamical behaviors of the chaotic controlled system are investigated in some detail. (c) 2002 American Institute of Physics. PMID- 12779564 TI - On creation of Hopf bifurcations in discrete-time nonlinear systems. AB - Bifurcation characteristics of a nonlinear system can be manipulated by small controls. In this paper, we present a control method to create Hopf bifurcations in discrete-time nonlinear systems. The critical conditions for the Hopf bifurcations are discussed. The center manifold method, normal form technique and the Iooss's Hopf bifurcation theory are employed in the derivation of the control gain. Numerical demonstration is provided. (c) 2002 American Institute of Physics. PMID- 12779566 TI - Introduction: Active chaotic flow. PMID- 12779565 TI - Arnold's method for asymptotic stability of steady inviscid incompressible flow through a fixed domain with permeable boundary. AB - The flow of an ideal fluid in a domain with a permeable boundary may be asymptotically stable. Here the permeability means that the fluid can flow into and out of the domain through some parts of the boundary. This permeability is a principal reason for the asymptotic stability. Indeed, the well-known conservation laws make the asymptotic stability of an inviscid flow impossible, if the usual no flux condition on a rigid wall (or on a free boundary) is employed. We study the stability problem using the direct Lyapunov method in the Arnold's form. We prove the linear and nonlinear Lyapunov stability of a two dimensional flow through a domain with a permeable boundary under Arnold's conditions. Under certain additional conditions, we amplify the linear result and prove the exponential decay of small disturbances. Here we employ the plan of the proof of the Barbashin-Krasovskiy theorem, established originally only for systems with a finite number of degrees of freedom. (c) 2002 American Institute of Physics. PMID- 12779567 TI - Chaotic stirring by a mesoscale surface-ocean flow. AB - The horizontal stirring properties of the flow in a region of the East Australian Current are calculated. A surface velocity field derived from remotely sensed data, using the maximum cross correlation method, is integrated to derive the distribution of the finite-time Lyapunov exponents. For the region studied (between latitudes 36 degrees S and 41 degrees S and longitudes 150 degrees E and 156 degrees E) the mean Lyapunov exponent during 1997 is estimated to be lambda( infinity )=4x10(-7) s(-1). This is in close agreement with the few other measurements of stirring rates in the surface ocean which are available. Recent theoretical results on the multifractal spectra of advected reactive tracers are applied to an analysis of a sea-surface temperature image of the study region. The spatial pattern seen in the image compares well with the pattern seen in an advected tracer with a first-order response to changes in surface forcing. The response timescale is estimated to be 20 days. (c) 2002 American Institute of Physics. PMID- 12779568 TI - Hyperbolic lines and the stratospheric polar vortex. AB - The necessary and sufficient conditions for Lagrangian hyperbolicity recently derived in the literature are reviewed in the light of older concepts of effective local rotation in strain coordinates. In particular, we introduce the simple interpretation of the necessary condition as a constraint on the local angular displacement in strain coordinates. These mathematically rigorous conditions are applied to the winter stratospheric circulation of the southern hemisphere, using analyzed wind data from the European Center for Medium-Range Weather Forecasts. Our results demonstrate that the sufficient condition is too strong and the necessary condition is too weak, so that both conditions fail to identify hyperbolic lines in the stratosphere. However a phenomenological, nonrigorous, criterion based on the necessary condition reveals the hyperbolic structure of the flow. Another (still nonrigorous) alternative is the finite-size Lyapunov exponent (FSLE) which is shown to produce good candidates for hyperbolic lines. In addition, we also tested the sufficient condition for Lagrangian ellipticity and found that it is too weak to detect elliptic coherent structures (ECS) in the stratosphere, of which the polar vortex is an obvious candidate. Yet, the FSLE method reveals a clear ECS-like barrier to mixing along the polar vortex edge. Further theoretical advancement is needed to explain the apparent success of nonrigorous methods, such as the FSLE approach, so as to achieve a sound kinematic understanding of chaotic mixing in the winter stratosphere and other geophysical flows. (c) 2002 American Institute of Physics. PMID- 12779569 TI - Structure analysis of turbulent mixing patterns in inert and reactive turbulent liquid flows. AB - The paper contains an extended summary of an invited plenary talk given at the Workshop on Active Chaos at the Los Alamos National Laboratory on 29-31 May 2001 by one of us (F.S.R.). (c) 2002 American Institute of Physics. PMID- 12779570 TI - Open problems in active chaotic flows: Competition between chaos and order in granular materials. AB - There are many systems where interaction among the elementary building blocks-no matter how well understood-does not even give a glimpse of the behavior of the global system itself. Characteristic for these systems is the ability to display structure without any external organizing principle being applied. They self organize as a consequence of synthesis and collective phenomena and the behavior cannot be understood in terms of the systems' constitutive elements alone. A simple example is flowing granular materials, i.e., systems composed of particles or grains. How the grains interact with each other is reasonably well understood; as to how particles move, the governing law is Newton's second law. There are no surprises at this level. However, when the particles are many and the material is vibrated or tumbled, surprising behavior emerges. Systems self-organize in complex patterns that cannot be deduced from the behavior of the particles alone. Self-organization is often the result of competing effects; flowing granular matter displays both mixing and segregation. Small differences in either size or density lead to flow-induced segregation and order; similar to fluids, noncohesive granular materials can display chaotic mixing and disorder. Competition gives rise to a wealth of experimental outcomes. Equilibrium structures, obtained experimentally in quasi-two-dimensional systems, display organization in the presence of disorder, and are captured by a continuum flow model incorporating collisional diffusion and density-driven segregation. Several open issues remain to be addressed. These include analysis of segregating chaotic systems from a dynamical systems viewpoint, and understanding three-dimensional systems and wet granular systems (slurries). General aspects of the competition between chaos-enhanced mixing and properties-induced de-mixing go beyond granular materials and may offer a paradigm for other kinds of physical systems. (c) 2002 American Institute of Physics. PMID- 12779571 TI - Autocatalytic reactions of phase distributed active particles. AB - We investigate the effect of asynchronism of autocatalytic reactions taking place in open hydrodynamical flows, by assigning a phase to each particle in the system to differentiate the timing of the reaction, while the reaction rate (periodicity) is kept unchanged. The chaotic saddle in the flow dynamics acts as a catalyst and enhances the reaction in the same fashion as in the case of a synchronous reaction that was studied previously, proving that the same type of nonlinear reaction kinetics is valid in the phase-distributed situation. More importantly, we show that, in a certain range of a parameter, the phenomenon of phase selection can occur, when a group of particles with a particular phase is favored over the others, thus occupying a larger fraction of the available space, or eventually leading to the extinction of the unfavored phases. We discuss the biological relevance of this latter phenomenon. (c) 2002 American Institute of Physics. PMID- 12779572 TI - Noise-induced enhancement of chemical reactions in nonlinear flows. AB - Motivated by the problem of ozone production in atmospheres of urban areas, we consider chemical reactions of the general type: A+B-->2C, in idealized two dimensional nonlinear flows that can generate Lagrangian chaos. Our aims differ from those in the existing work in that we address the role of transient chaos versus sustained chaos and, more importantly, we investigate the influence of noise. We find that noise can significantly enhance the chemical reaction in a resonancelike manner where the product of the reaction becomes maximum at some optimal noise level. We also argue that chaos may not be a necessary condition for the observed resonances. A physical theory is formulated to understand the resonant behavior. (c) 2002 American Institute of Physics. PMID- 12779573 TI - Chaotic mixing induced transitions in reaction-diffusion systems. AB - We study the evolution of a localized perturbation in a chemical system with multiple homogeneous steady states, in the presence of stirring by a fluid flow. Two distinct regimes are found as the rate of stirring is varied relative to the rate of the chemical reaction. When the stirring is fast localized perturbations decay towards a spatially homogeneous state. When the stirring is slow (or fast reaction) localized perturbations propagate by advection in form of a filament with a roughly constant width and exponentially increasing length. The width of the filament depends on the stirring rate and reaction rate but is independent of the initial perturbation. We investigate this problem numerically in both closed and open flow systems and explain the results using a one-dimensional "mean strain" model for the transverse profile of the filament that captures the interplay between the propagation of the reaction-diffusion front and the stretching due to chaotic advection. (c) 2002 American Institute of Physics. PMID- 12779574 TI - Surface quasigeostrophic turbulence: The study of an active scalar. AB - We study the statistical and geometrical properties of the potential temperature (PT) field in the surface quasigeostrophic (SQG) system of equations. In addition to extracting information in a global sense via tools such as the power spectrum, the g-beta spectrum, and the structure functions we explore the local nature of the PT field by means of the wavelet transform method. The primary indication is that an initially smooth PT field becomes rough (within specified scales), though in a qualitatively sparse fashion. Similarly, initially one-dimensional iso-PT contours (i.e., PT level sets) are seen to acquire a fractal nature. Moreover, the dimensions of the iso-PT contours satisfy existing analytical bounds. The expectation that the roughness will manifest itself in the singular nature of the gradient fields is confirmed via the multifractal nature of the dissipation field. Following earlier work on the subject, the singular and oscillatory nature of the gradient field is investigated by examining the scaling of a probability measure and a sign singular measure, respectively. A physically motivated derivation of the relations between the variety of scaling exponents is presented, the aim being to bring out some of the underlying assumptions which seem to have gone unnoticed in previous presentations. Apart from concentrating on specific properties of the SQG system, a broader theme of the paper is a comparison of the diagnostic inertial range properties of the SQG system with both the two- and three-dimensional Euler equations. (c) 2002 American Institute of Physics. PMID- 12779575 TI - Water vapor as an active scalar in tropical atmospheric dynamics. AB - Water vapor is a constituent of the tropical atmosphere which, though to a significant extent locally controlled by vertical advection, precipitation, and surface evaporation, is also affected by horizontal advection. Water vapor affects the flow in turn, because a humid atmosphere supports deep, precipitating convection more readily than a dry atmosphere. Precipitation heats the atmosphere, and this heating drives the flow. Water vapor is thus a dynamically active constituent. Simplifications to the primitive equations of dynamical meteorology, based on the so-called weak temperature gradient approximation, are presented which highlight this behavior. The weak temperature gradient approximation is valid on large scales near the equator. It eliminates gravity waves, leaving only balanced dynamics, though the fundamental balance occurs in the temperature rather than the momentum equation (as is customary in most balance models of geophysical fluid dynamics). The dynamical role of water vapor is examined in a couple of idealized contexts, where either the vertical or horizontal structure of the flow is severely simplified. (c) 2002 American Institute of Physics. PMID- 12779576 TI - Metabolic network dynamics in open chaotic flow. AB - We have analyzed the dynamics of metabolically coupled replicators in open chaotic flows. Replicators contribute to a common metabolism producing energy rich monomers necessary for replication. The flow and the biological processes take place on a rectangular grid. There can be at most one molecule on each grid cell, and replication can occur only at localities where all the necessary replicators (metabolic enzymes) are present within a certain neighborhood distance. Due to this finite metabolic neighborhood size and imperfect mixing along the fractal filaments produced by the flow, replicators can coexist in this fluid system, even though coexistence is impossible in the mean-field approximation of the model. We have shown numerically that coexistence mainly depends on the metabolic neighborhood size, the kinetic parameters, and the number of replicators coupled through metabolism. Selfish parasite replicators cannot destroy the system of coexisting metabolic replicators, but they frequently remain persistent in the system. (c) 2002 American Institute of Physics. PMID- 12779577 TI - Small-scale structure of nonlinearly interacting species advected by chaotic flows. AB - We study the spatial patterns formed by interacting biological populations or reacting chemicals under the influence of chaotic flows. Multiple species and nonlinear interactions are explicitly considered, as well as cases of smooth and nonsmooth forcing sources. The small-scale structure can be obtained in terms of characteristic Lyapunov exponents of the flow and of the chemical dynamics. Different kinds of morphological transitions are identified. Numerical results from a three-component plankton dynamics model support the theory, and they serve also to illustrate the influence of asymmetric couplings. (c) 2002 American Institute of Physics. PMID- 12779578 TI - Front speed enhancement in cellular flows. AB - The problem of front propagation in a stirred medium is addressed in the case of cellular flows in three different regimes: slow reaction, fast reaction and geometrical optics limit. It is well known that a consequence of stirring is the enhancement of front speed with respect to the nonstirred case. By means of numerical simulations and theoretical arguments we describe the behavior of front speed as a function of the stirring intensity, U. For slow reaction, the front propagates with a speed proportional to U(1/4), conversely for fast reaction the front speed is proportional to U(3/4). In the geometrical optics limit, the front speed asymptotically behaves as U/ln U. (c) 2002 American Institute of Physics. PMID- 12779579 TI - Noise- and inertia-induced inhomogeneity in the distribution of small particles in fluid flows. AB - The dynamics of small spherical neutrally buoyant particulate impurities immersed in a two-dimensional fluid flow are known to lead to particle accumulation in the regions of the flow in which vorticity dominates over strain, provided that the Stokes number of the particles is sufficiently small. If the flow is viewed as a Hamiltonian dynamical system, it can be seen that the accumulations occur in the nonchaotic parts of the phase space: the Kolmogorov-Arnold-Moser tori. This has suggested a generalization of these dynamics to Hamiltonian maps, dubbed a bailout embedding. In this paper we use a bailout embedding of the standard map to mimic the dynamics of neutrally buoyant impurities subject not only to drag but also to fluctuating forces modeled as white noise. We find that the generation of inhomogeneities associated with the separation of particle from fluid trajectories is enhanced by the presence of noise, so that they appear in much broader ranges of the Stokes number than those allowing spontaneous separation. (c) 2002 American Institute of Physics. PMID- 12779581 TI - The effect of Lagrangian chaos on locking bifurcations in shear flows. AB - The effect of an externally imposed perturbation on an unstable or weakly stable shear flow is investigated, with a focus on the role of Lagrangian chaos in the bifurcations that occur. The external perturbation is at rest in the laboratory frame and can form a chain of resonances or cat's eyes where the initial velocity v(x0)(y) vanishes. If in addition the shear profile is unstable or weakly stable to a Kelvin-Helmholtz instability, for a certain amplitude of the external perturbation there can be an unlocking bifurcation to a nonlinear wave resonant around a different value of y, with nonzero phase velocity. The interaction of the propagating nonlinear wave with the external perturbation leads to Lagrangian chaos. We discuss results based on numerical simulations for different amplitudes of the external perturbation. The response to the external perturbation is strong, apparently because of non-normality of the linear operator, and the unlocking bifurcation is hysteretic. The results indicate that the observed Lagrangian chaos is responsible for a second bifurcation occurring at larger external perturbation, locking the wave to the wall. This bifurcation is nonhysteretic. The mechanism by which the chaos leads to locking in this second bifurcation is by means of chaotic advective transport of momentum from one chain of resonances to the other (Reynolds stress) and momentum transport to the vicinity of the wall via chaotic scattering. These results suggest that locking of waves in rotating tank experiments in the presence of two unstable modes is due to a similar process. (c) 2002 American Institute of Physics. PMID- 12779580 TI - Coherent structures and self-consistent transport in a mean field Hamiltonian model. AB - A study of coherent structures and self-consistent transport is presented in the context of a Hamiltonian mean field, single wave model. The model describes the weakly nonlinear dynamics of marginally stable plasmas and fluids, and it is related to models of systems with long-range interactions in statistical mechanics. In plasma physics the model applies to the interaction of electron "holes" and electron "clumps," which are depletions and excesses of phase-space electron density with respect to a fixed background. In fluid dynamics the system describes the interaction of vortices with positive and negative circulation in a two-dimensional background shear flow. Numerical simulations in the finite-N and in the N--> infinity kinetic limit (where N is the number of particles) show the existence of coherent, rotating dipole states. We approximate the dipole as two "macroparticles" (one hole and one clump) and consider the N=2 limit of the model. We show that this limit has a family of symmetric, rotating integrable solutions described by a one-degree-of-freedom nontwist Hamiltonian. A perturbative solution of the nontwist Hamiltonian provides an accurate description of the mean field and rotation period of the dipole. The coherence of the dipole is explained in terms of a parametric resonance between the rotation frequency of the macroparticles and the oscillation frequency of the self consistent mean field. This resonance creates islands of integrability that shield the dipole from regions of chaotic transport. For a class of initial conditions, the mean field exhibits an elliptic-hyperbolic bifurcation that leads to the filamentation, chaotic mixing and eventual destruction of the dipole. (c) 2002 American Institute of Physics. PMID- 12779582 TI - Lagrangian averages, averaged Lagrangians, and the mean effects of fluctuations in fluid dynamics. AB - We begin by placing the generalized Lagrangian mean (GLM) equations for a compressible adiabatic fluid into the Euler-Poincare (EP) variational framework of fluid dynamics, for an averaged Lagrangian. This is the Lagrangian averaged Euler-Poincare (LAEP) theorem. Next, we derive a set of approximate small amplitude GLM equations (glm equations) at second order in the fluctuating displacement of a Lagrangian trajectory from its mean position. These equations express the linear and nonlinear back-reaction effects on the Eulerian mean fluid quantities by the fluctuating displacements of the Lagrangian trajectories in terms of their Eulerian second moments. The derivation of the glm equations uses the linearized relations between Eulerian and Lagrangian fluctuations, in the tradition of Lagrangian stability analysis for fluids. The glm derivation also uses the method of averaged Lagrangians, in the tradition of wave, mean flow interaction. Next, the new glm EP motion equations for incompressible ideal fluids are compared with the Euler-alpha turbulence closure equations. An alpha model is a GLM (or glm) fluid theory with a Taylor hypothesis closure. Such closures are based on the linearized fluctuation relations that determine the dynamics of the Lagrangian statistical quantities in the Euler-alpha equations. Thus, by using the LAEP theorem, we bridge between the GLM equations and the Euler-alpha closure equations, through the small-amplitude glm approximation in the EP variational framework. We conclude by highlighting a new application of the GLM, glm, and alpha-model results for Lagrangian averaged ideal magnetohydrodynamics. (c) 2002 American Institute of Physics. PMID- 12779583 TI - Food chain chaos due to Shilnikov's orbit. AB - Assume that the reproduction rate ratio zeta of the predator over the prey is sufficiently small in a basic tri-trophic food chain model. This assumption translates the model into a singularly perturbed system of two time scales. It is demonstrated, as a sequel to the earlier paper of Deng [Chaos 11, 514-525 (2001)], that at the singular limit zeta=0, a singular Shilnikov's saddle-focus homoclinic orbit can exist as the reproduction rate ratio epsilon of the top predator over the predator is greater than a modest value epsilon(0). The additional conditions under which such a singular orbit may occur are also explicitly given. (c) 2002 American Institute of Physics. PMID- 12779584 TI - Degenerate resonances in Hamiltonian systems with 3/2 degrees of freedom. AB - Hamiltonian systems with 3/2 degrees of freedom close to autonomous systems are considered. Special attention is focused on the case of degenerate resonances. In this case, an averaged system in the first approximation reduces to an area preserving mapping of a cylinder whose rotation number is a nonmonotonic function of the action variable. Behavior of the trajectories of such a map is similar to that of the trajectories of a Poincare map. Three regions: B(+/-) in the upper and lower parts of the cylinder and an additional region A which contains separatrices of fixed points for the corresponding resonance are distinguished on the cylinder. It is shown that there is a nonempty set of initial points corresponding to walking trajectories in B(+/-) and, hence, there are no closed invariant curves that are homotopically nontrivial on the cylinder. Cells limited by a "stochastic network" can exist in region A. The number of cells is the greater the higher the order of degeneration of the resonance. Possible types of orbit behavior in region A are described. (c) 2002 American Institute of Physics. PMID- 12779585 TI - Investigating nonlinear dynamics from time series: The influence of symmetries and the choice of observables. AB - When a dynamical system is investigated from a time series, one of the most challenging problems is to obtain a model that reproduces the underlying dynamics. Many papers have been devoted to this problem but very few have considered the influence of symmetries in the original system and the choice of the observable. Indeed, it is well known that there are usually some variables that provide a better representation of the underlying dynamics and, consequently, a global model can be obtained with less difficulties starting from such variables. This is connected to the problem of observing the dynamical system from a single time series. The roots of the nonequivalence between the dynamical variables will be investigated in a more systematic way using previously defined observability indices. It turns out that there are two important ingredients which are the complexity of the coupling between the dynamical variables and the symmetry properties of the original system. As will be mentioned, symmetries and the choice of observables also has important consequences in other problems such as synchronization of nonlinear oscillators. (c) 2002 American Institute of Physics. PMID- 12779586 TI - Chaos control by using Motor Maps. AB - In this paper a new method for chaos control is proposed, consisting of an unsupervised neural network, namely a Motor Map. In particular a feedback entrainment scheme is adopted: a chaotic system with a given parameter set generates the reference trajectory for another chaotic system with different parameters to be controlled: the Motor Map is required to provide the appropriate time-varying gain value for the feedback signal. The state of the controlled system is considered as input to the Motor Map. Particular efforts have been paid to the feasibility of the implementation. Indeed, the simulations performed have been oriented to design a Motor Map suitable for an hardware realization, thus some restrictive hypotheses, such as for example a low number of neurons, have been assumed. A huge number of simulations has been carried out by considering as system to be controlled a Double Scroll Chua Attractor as well as other chaotic attractors. Several reference trajectories have also been considered: a limit cycle generated by a Chua's circuit with different parameters values, a double scroll Chua attractor, a chaotic attractor of the family of the Chua's circuit attractors. In all the simulations instead of controlling the whole state space, only two state variables have been fed back. Good results in terms of settling time (namely, the period in which the map learns the control task) and steady state errors have been obtained with a few neurons. The Motor Map based adaptive controller offers high performances, specially in the case when the reference trajectory is switched into another one. In this case, a specialization of the neurons constituting the Motor Map is observed: while a group of neurons learns the appropriate control law for a reference trajectory, another group specializes itself to control the system when the other trajectory is used as a reference. A discrete components electronic realization of the Motor Map is presented and experimental results confirming the simulation results are shown. (c) 2002 American Institute of Physics. PMID- 12779587 TI - The stability of stationary rotation of a regular vortex polygon. AB - This paper is devoted to the Lord Kelvin's (1878) problem on stability of the stationary rotation of the system of n equal vortices located in the vertices of a regular n-gon. During the last decades this problem again became actual in connection with the investigation of point vortices in liquid helium and electron columns in plasma physics. This regime is described by the explicit solution of the Kirchhoff equations. The corresponding eigenvalue problem for the linearization matrix can be also decided explicitly. This was used in the works of Thomson (1883) and Havelock (1931) to obtain exhaustive results on the linear stability. Kurakin (1994) proved that for n/=8 it is unstable. We also present the general theory of stationary motions of a dynamical system with symmetry group. The definitions of stability and instability are necessary to modify in the specific case of stationary regimes. We do not assume that the system is conservative. Thus, the results can be applied not only to various stationary regimes of an ideal fluid flows but, for instance, also to motions of viscous fluids. (c) 2002 American Institute of Physics. PMID- 12779588 TI - Recurrence plots and unstable periodic orbits. AB - A recurrence plot is a two-dimensional visualization technique for sequential data. These plots are useful in that they bring out correlations at all scales in a manner that is obvious to the human eye, but their rich geometric structure can make them hard to interpret. In this paper, we suggest that the unstable periodic orbits embedded in a chaotic attractor are a useful basis set for the geometry of a recurrence plot of those data. This provides not only a simple way to locate unstable periodic orbits in chaotic time-series data, but also a potentially effective way to use a recurrence plot to identify a dynamical system. (c) 2002 American Institute of Physics. PMID- 12779589 TI - Onset of chaotic dynamics in a ball mill: Attractors merging and crisis induced intermittency. AB - In mechanical treatment carried out by ball milling, powder particles are subjected to repeated high-energy mechanical loads which induce heavy plastic deformations together with fracturing and cold-welding events. Owing to the continuous defect accumulation and interface renewal, both structural and chemical transformations occur. The nature and the rate of such transformations have been shown to depend on variables, such as impact velocity and collision frequency that depend, in turn, on the whole dynamics of the system. The characterization of the ball dynamics under different impact conditions is then to be considered a necessary step in order to gain a satisfactory control of the experimental set up. In this paper we investigate the motion of a ball in a milling device. Since the ball motion is governed by impulsive forces acting during each collision, no analytical expression for the complete ball trajectory can be obtained. In addition, mechanical systems exhibiting impacts are strongly nonlinear due to sudden changes of velocities at the instant of impact. Many different types of periodic and chaotic impact motions exist indeed even for simple systems with external periodic excitation forces. We present results of the analysis on the ball trajectory, obtained from a suitable numerical model, under growing degree of impact elasticity. A route to high dimensional chaos is obtained. Crisis and attractors merging are also found. (c) 2002 American Institute of Physics. PMID- 12779590 TI - Stability of active systems with a spatially periodic activity: Analysis of a simple model and application to the boiling crisis problem. AB - We investigate theoretically the possibility to control the transition between two metastable states in reactive systems by imposing a spatial modulation. In particular, we consider the technologically very important case of the transition between the low temperature (nucleate boiling) and the high temperature (film boiling) phases of boiling of a liquid over a heat generating element, also known as the boiling crisis. With the help of a simplified model, we demonstrate that the dangerous regime where the high temperature phase invades the whole system requires a larger heat power in a periodically spatially modulated system, than in a uniform system. The possibility that a local perturbation, such as a small gas bubble, may induce locally a transition to the film boiling state is also considered. We show that the transition to the film boiling regime is hindered in a spatially periodic system. (c) 2002 American Institute of Physics. PMID- 12779591 TI - Sensitivity of ray travel times. AB - Ray in a waveguide can be considered as a trajectory of the corresponding Hamiltonian system, which appears to be chaotic in a nonuniform environment. From the experimental and practical viewpoints, the ray travel time is an important characteristic that, in some way, involves an information about the waveguide condition. It is shown that the ray travel time as a function of the initial momentum and propagation range in the unperturbed waveguide displays a scaling law. Some properties of the ray travel time predicted by this law still persist in periodically nonuniform waveguides with chaotic ray trajectories. As examples we consider few models with special attention to the underwater acoustic waveguide. It is demonstrated for a deep ocean propagation model that even under conditions of ray chaos the ray travel time is determined, to a considerable extent, by the coordinates of the ray endpoints and the number of turning points, i.e., by a topology of the ray path. We show how the closeness of travel times for rays with equal numbers of turning points reveals itself in ray travel time dependencies on the starting momentum and on the depth of the observation point. It has been shown that the same effect is associated with the appearance of the gap between travel times of chaotic and regular rays. The manifestation of the stickiness (the presence of such parts in a chaotic trajectory where the latter exhibits an almost regular behavior) in ray travel times is discussed. (c) 2002 American Institute of Physics. PMID- 12779592 TI - Cookbook asymptotics for spiral and scroll waves in excitable media. AB - Algebraic formulas predicting the frequencies and shapes of waves in a reaction diffusion model of excitable media are presented in the form of four recipes. The formulas themselves are based on a detailed asymptotic analysis (published elsewhere) of the model equations at leading order and first order in the asymptotic parameter. The importance of the first order contribution is stressed throughout, beginning with a discussion of the Fife limit, Fife scaling, and Fife regime. Recipes are given for spiral waves and detailed comparisons are presented between the asymptotic predictions and the solutions of the full reaction diffusion equations. Recipes for twisted scroll waves with straight filaments are given and again comparisons are shown. The connection between the asymptotic results and filament dynamics is discussed, and one of the previously unknown coefficients in the theory of filament dynamics is evaluated in terms of its asymptotic expansion. (c) 2002 American Institute of Physics. PMID- 12779593 TI - Chaos in the relativistic two-electron atom. AB - Chaotic autoionization of the relativistic two-electron atom is investigated. A theoretical analysis of chaotic dynamics of the relativistic outer electron under the periodic perturbation due to the inner electron, assumed to be on a circular orbit, based on the Chirikov criterion, is given. The diffusion coefficient, the ionization rate, and time are calculated. (c) 2002 American Institute of Physics. PMID- 12779594 TI - Transversal homoclinic orbits in a transiently chaotic neural network. AB - We study the existence of snap-back repellers, hence the existence of transversal homoclinic orbits in a discrete-time neural network. Chaotic behaviors for the network system in the sense of Li and Yorke or Marotto can then be concluded. The result is established by analyzing the structures of the system and allocating suitable parameters in constructing the fixed points and their pre-images for the system. The investigation provides a theoretical confirmation on the scenario of transient chaos for the system. All the parameter conditions for the theory can be examined numerically. The numerical ranges for the parameters which yield chaotic dynamics and convergent dynamics provide significant information in the annealing process in solving combinatorial optimization problems using this transiently chaotic neural network. (c) 2002 American Institute of Physics. PMID- 12779595 TI - Winnerless competition between sensory neurons generates chaos: A possible mechanism for molluscan hunting behavior. AB - In the presence of prey, the marine mollusk Clione limacina exhibits search behavior, i.e., circular motions whose plane and radius change in a chaotic-like manner. We have formulated a dynamical model of the chaotic hunting behavior of Clione based on physiological in vivo and in vitro experiments. The model includes a description of the action of the cerebral hunting interneuron on the receptor neurons of the gravity sensory organ, the statocyst. A network of six receptor model neurons with Lotka-Volterra-type dynamics and nonsymmetric inhibitory interactions has no simple static attractors that correspond to winner take all phenomena. Instead, the winnerless competition induced by the hunting neuron displays hyperchaos with two positive Lyapunov exponents. The origin of the chaos is related to the interaction of two clusters of receptor neurons that are described with two heteroclinic loops in phase space. We hypothesize that the chaotic activity of the receptor neurons can drive the complex behavior of Clione observed during hunting. (c) 2002 American Institute of Physics. PMID- 12779596 TI - Collective chaos synchronization of pairs of modes in a chaotic three-mode laser. AB - We study chaos synchronization experimentally in a modulated globally coupled three-mode laser with different modal gains subjected to self-mixing Doppler shifted feedback, which can apply the loss modulation to individual modes at Doppler-shift frequencies. Depending on the pump power, different forms of collective chaos synchronizations were found to appear when the laser was modulated at the highest relaxation oscillation frequency, reflecting the change in cross-saturation coefficient among modes. In the present experiment, each pair of modes exhibited phase, lag, or generalized synchronization collectively according to the inherent antiphase dynamics, where these types of synchronization have already been demonstrated in two coupled chaotic oscillators in different physical systems. Information flows among oscillating modes which are established in different forms of collective chaos synchronizations were characterized by information-circulation analysis of the experimental time series. (c) 2002 American Institute of Physics. PMID- 12779597 TI - Characterization of finite-time Lyapunov exponents and vectors in two-dimensional turbulence. AB - This paper discusses the application of Lyapunov theory in chaotic systems to the dynamics of tracer gradients in two-dimensional flows. The Lyapunov theory indicates that more attention should be given to the Lyapunov vector orientation. Moreover, the properties of Lyapunov vectors and exponents are explained in light of recent results on tracer gradients dynamics. Differences between the different Lyapunov vectors can be interpreted in terms of competition between the effects of effective rotation and strain. Also, the differences between backward and forward vectors give information on the local reversibility of the tracer gradient dynamics. A numerical simulation of two-dimensional turbulence serves to highlight these points and the spatial distribution of finite time Lyapunov exponents is also discussed in relation to stirring properties. (c) 2002 American Institute of Physics. PMID- 12779598 TI - Reversible maps in two-degrees of freedom Hamiltonian systems. AB - It has been shown that a sub-class of two-degrees of freedom Hamiltonian systems possesses a reversing symmetry discovered by Birkhoff in the restricted problem of three bodies. This mixed space-time reversing symmetry, which is different from the classical time reversal symmetry, can be shared by time-reversible as well as time-irreversible systems. Examples of time-irreversible systems which possess this reversing symmetry are the restricted problem of three bodies as shown by Birkhoff in 1915, and a special case of the motion of a rigid body with a fixed point discussed in this paper. If a Hamiltonian system possesses this Birkhoff reversing symmetry, then there exists a surface of section for which the corresponding Poincare map is Birkhoff-reversible. The Birkhoff-reversibility of this map may be used to study its global dynamics such as the locations and the distribution of the stable and unstable periodic points, the distribution of stable and chaotic regions, and the identification of the scattering regions. (c) 2002 American Institute of Physics. PMID- 12779599 TI - Instabilities and spatio-temporal chaos of long-wave hexagon patterns in rotating Marangoni convection. AB - We consider surface-tension driven convection in a rotating fluid layer. For nearly insulating boundary conditions we derive a long-wave equation for the convection planform. Using a Galerkin method and direct numerical simulations we study the stability of the steady hexagonal patterns with respect to general side band instabilities. In the presence of rotation, steady and oscillatory instabilities are identified. One of them leads to stable, homogeneously oscillating hexagons. For sufficiently large rotation rates the stability balloon closes, rendering all steady hexagons unstable and leading to spatio-temporal chaos. (c) 2002 American Institute of Physics. PMID- 12779600 TI - A hierarchy of coupled maps. AB - A large number of logistic maps are coupled together as a mathematical metaphor for complex natural systems with hierarchical organization. The elementary maps are first collected into globally coupled lattices. These lattices are then coupled together in a hierarchical way to form a system with many degrees of freedom. We summarize the behavior of the individual blocks, and then explore the dynamics of the hierarchy. We offer some ideas that guide our understanding of this type of system. (c) 2002 American Institute of Physics. PMID- 12779601 TI - Introduction: Mapping and control of complex cardiac arrhythmias. AB - This paper serves as an introduction to the Focus Issue on mapping and control of complex cardiac arrhythmias. We first introduce basic concepts of cardiac electrophysiology and describe the main clinical methods being used to treat arrhythmia. We then provide a brief summary of the main themes contained in the articles in this Focus Issue. In recent years there have been important advances in the ability to map the spread of excitation in intact hearts and in laboratory settings. This work has been combined with simulations that use increasingly realistic geometry and physiology. Waves of excitation and contraction in the heart do not always propagate with constant velocity but are often subject to instabilities that may lead to fluctuations in velocity and cycle time. Such instabilities are often treated best in the context of simple one- or two dimensional geometries. An understanding of the mechanisms of propagation and wave stability is leading to the implementation of different stimulation protocols in an effort to modify or eliminate abnormal rhythms. (c) 2002 American Institute of Physics. PMID- 12779602 TI - Anatomic determinants of atrial arrhythmias: New insights from three-dimensional mapping. AB - The recent development of new technologies for in vivo three-dimensional mapping of arrhythmias has enabled a better understanding of the richness of intracardiac anatomy and the relationship between anatomy and arrhythmogenesis. In the present manuscript we review two new technologies for in vivo mapping of atrial arrhythmias and explores the degree to which the anatomic complexity they reveal is important in determining the physiology of both focal and macroreentrant atrial tachycardias. These observations highlight the importance of including sufficient anatomic detail in modeling studies aimed at elucidating the pathophysiology of atrial arrhythmias. (c) 2002 American Institute of Physics. PMID- 12779603 TI - Propagation through heterogeneous substrates in simple excitable media models. AB - The interaction of waves and obstacles is simulated by adding heterogeneities to a FitzHugh-Nagumo model and a cellular automata model. The cellular automata model is formulated to account for heterogeneities by modelling the interaction between current sources and current sinks. In both models, wave fronts propagate if the size of the heterogeneities is small, and block if the size of the heterogeneities is large. For intermediate values, wave fronts break up into numerous spiral waves. The theoretical models give insights concerning spiral wave formation in heterogeneous excitable media. (c) 2002 American Institute of Physics. PMID- 12779604 TI - Study of atrial arrhythmias in a computer model based on magnetic resonance images of human atria. AB - The maintenance of multiple wavelets appears to be a consistent feature of atrial fibrillation (AF). In this paper, we investigate possible mechanisms of initiation and perpetuation of multiple wavelets in a computer model of AF. We developed a simplified model of human atria that uses an ionic-based membrane model and whose geometry is derived from a segmented magnetic resonance imaging data set. The three-dimensional surface has a realistic size and includes obstacles corresponding to the location of major vessels and valves, but it does not take into account anisotropy. The main advantage of this approach is its ability to simulate long duration arrhythmias (up to 40 s). Clinically relevant initiation protocols, such as single-site burst pacing, were used. The dynamics of simulated AF were investigated in models with different action potential durations and restitution properties, controlled by the conductance of the slow inward current in a modified Luo-Rudy model. The simulation studies show that (1) single-site burst pacing protocol can be used to induce wave breaks even in tissue with uniform membrane properties, (2) the restitution-based wave breaks in an atrial model with realistic size and conduction velocities are transient, and (3) a significant reduction in action potential duration (even with apparently flat restitution) increases the duration of AF. (c) 2002 American Institute of Physics. PMID- 12779605 TI - Development of a computer algorithm for the detection of phase singularities and initial application to analyze simulations of atrial fibrillation. AB - Atrial fibrillation (AF) is a common cardiac arrhythmia, but its mechanisms are incompletely understood. The identification of phase singularities (PSs) has been used to define spiral waves involved in maintaining the arrhythmia, as well as daughter wavelets. In the past, PSs have often been identified manually. Automated PS detection algorithms have been described previously, but when we attempted to apply a previously developed algorithm we experienced problems with false positives that made the results difficult to use directly. We therefore developed a tool for PS identification that uses multiple strategies incorporating both image analysis and mathematical convolution for automated detection with optimized sensitivity and specificity, followed by manual verification. The tool was then applied to analyze PS behavior in simulations of AF maintained in the presence of spatially distributed acetylcholine effects in cell grids of varying size. These analyses indicated that in almost all cases, a single PS lasted throughout the simulation, corresponding to the central-core tip of a single spiral wave that maintained AF. The sustained PS always localized to an area of low acetylcholine concentration. When the grid became very small and no area of low acetylcholine concentration was surrounded by zones of higher concentration, AF could not be sustained. The behavior of PSs and the mechanisms of AF were qualitatively constant over an 11.1-fold range of atrial grid size, suggesting that the classical emphasis on tissue size as a primary determinant of fibrillatory behavior may be overstated. (c) 2002 American Institute of Physics. PMID- 12779606 TI - Wave front fragmentation due to ventricular geometry in a model of the rabbit heart. AB - The role of the heart's complex shape in causing the fragmentation of activation wave fronts characteristic of ventricular fibrillation (VF) has not been well studied. We used a finite element model of cardiac propagation capable of simulating functional reentry on curved two-dimensional surfaces to test the hypothesis that uneven surface curvature can cause local propagation block leading to proliferation of reentrant wave fronts. We found that when reentry was induced on a flat sheet, it rotated in a repeatable meander pattern without breaking up. However, when a model of the rabbit ventricles was formed from the same medium, reentrant wave fronts followed complex, nonrepeating trajectories. Local propagation block often occurred when wave fronts propagated across regions where the Gaussian curvature of the surface changed rapidly. This type of block did not occur every time wave fronts crossed such a region; rather, it only occurred when the wave front was very close behind the previous wave in the cycle and was therefore propagating into relatively inexcitable tissue. Close wave front spacing resulted from nonstationary reentrant propagation. Thus, uneven surface curvature and nonstationary reentrant propagation worked in concert to produce wave front fragmentation and complex activation patterns. None of the factors previously thought to be necessary for local propagation block (e.g., heterogeneous refractory period, steep action potential duration restitution) were present. We conclude that the complex geometry of the heart may be an important determinant of VF activation patterns. (c) 2002 American Institute of Physics. PMID- 12779607 TI - Stability conditions for the traveling pulse: Modifying the restitution hypothesis. AB - As a simple model of reentry, we use a general FitzHugh-Nagumo model on a ring (in the singular limit) to build an understanding of the scope of the restitution hypothesis. It has already been shown that for a traveling pulse solution with a phase wave back, the restitution hypothesis gives the correct stability condition. We generalize this analysis to include the possibility of a pulse with a triggered wave back. Calculating the linear stability condition for such a system, we find that the restitution hypothesis, which depends only on action potential duration restitution, can be extended to a more general condition that includes dependence on conduction velocity restitution as well as two other parameters. This extension amounts to unfolding the original bifurcation described in the phase wave back case which was originally understood to be a degenerate bifurcation. In addition, we demonstrate that dependence of stability on the slope of the restitution curve can be significantly modified by the sensitivity to other parameters (including conduction velocity restitution). We provide an example in which the traveling pulse is stable despite a steep restitution curve. (c) 2002 American Institute of Physics. PMID- 12779608 TI - Wave propagation in an excitable medium with a negatively sloped restitution curve. AB - Recent experimental studies show that the restitution curve of cardiac tissue can have a negative slope. We study how the negative slope of the restitution curve can influence basic processes in excitable media, such as periodic forcing of an excitable cell, circulation of a pulse in a ring, and spiral wave rotation in two dimensions. We show that negatively sloped restitution curve can result in instabilities if the slope of the restitution curve is steeper than -1 and report different manifestations of this instability. (c) 2002 American Institute of Physics. PMID- 12779609 TI - Triggered alternans in an ionic model of ischemic cardiac ventricular muscle. AB - It has been known for several decades that electrical alternans occurs during myocardial ischemia in both clinical and experimental work. There are a few reports showing that this alternans can be triggered into existence by a premature ventricular contraction. Detriggering of alternans by a premature ventricular contraction, as well as pause-induced triggering and detriggering, have also been reported. We conduct a search for triggered alternans in an ionic model of ischemic ventricular muscle in which alternans has been described recently: a one-dimensional cable of length 3 cm, containing a central ischemic zone 1 cm long, with 1 cm segments of normal (i.e., nonischemic) tissue at each end. We use a modified form of the Luo-Rudy [Circ. Res. 68, 1501-1526 (1991)] ionic model to represent the ventricular tissue, modeling the effect of ischemia by raising the external potassium ion concentration ([K(+)](o)) in the central ischemic zone. As [K(+)](o) is increased at a fixed pacing cycle length of 400 ms, there is first a transition from 1:1 rhythm to alternans or 2:2 rhythm, and then a transition from 2:2 rhythm to 2:1 block. There is a range of [K(+)](o) over which there is coexistence of 1:1 and 2:2 rhythms, so that dropping a stimulus from the periodic drive train during 1:1 rhythm can result in the conversion of 1:1 to 2:2 rhythm. Within the bistable range, the reverse transition from 2:2 to 1:1 rhythm can be produced by injection of a well-timed extrastimulus. Using a stimulation protocol involving delivery of pre- and post mature stimuli, we derive a one-dimensional map that captures the salient features of the results of the cable simulations, i.e., the {1:1-->2:2-->2:1} transitions with {1:1<-->2:2} bistability. This map uses a new index of the global activity in the cable, the normalized voltage integral. Finally, we put forth a simple piecewise linear map that replicates the {1:1<-->2:2} bistability observed in the cable simulations and in the normalized voltage integral map. (c) 2002 American Institute of Physics. PMID- 12779610 TI - Interplay of ionic and structural heterogeneity on functional action potential duration gradients: Implications for arrhythmogenesis. AB - Action potential duration (APD) dispersion in the heart is governed by the underlying cellular architecture and the spatial distribution of the membrane properties. Understanding the contribution of each factor is important in designing more effective methods for the control of arrhythmias. Recent experimental studies have shown that the insertion of structural barriers in ionically heterogeneous tissue facilitates the formation of unidirectional block and discordant alternans. In this work, computational modeling is used to examine the effect of internal obstacles on the formation of functional APD gradients in ionically heterogeneous tissue. Intrinsic APD differences are introduced by assigning two discrete cell types to each half of a square domain. The combined effect of structural and ionic heterogeneities is shown to produce gradients in APD that are oblique to both the intrinsic gradients in APD and the physical boundary. Simulation results are presented that show that the magnitude and spatial extent of the subsequent APD gradients are modulated by the size and orientation of the obstacle, the degree of anisotropy, and the location of the pacing site. Long, thin internal obstacles are found to produce the greatest dispersion in APD. The combination of internal obstacles and ionic heterogeneities is shown to produce a substrate for re-entrant excitation following a pair of near threshold point stimuli. (c) 2002 American Institute of Physics. PMID- 12779611 TI - A primary mechanism for spiral wave meandering. AB - The stability and dynamics of spiral wave meandering were studied by examining the behavior of small perturbations to a steadily rotating action potential wave. The disturbances responsible for meandering were found to be generated through an interaction between the unstable local linear dynamics characteristic of the action potential trailing edge near the core and perturbations existing in the region immediately behind this edge. Significantly, for the cases studied, neither wavefront curvature nor head-tail interactions were involved in this process. Study of the generation mechanism using a series of representative mathematical models and computer experiments led to the prediction that the following features of rotating action potentials render them more susceptible to meandering: (1) proximity of the wave tip to the center of rotation, (2) wider action potential leading and trailing edges, and (3) slower wave rotation speeds. Variation of basic tissue properties, including firing threshold potentials and excitability above threshold, affected these properties, and those of the perturbation dynamics, in several ways, producing both stabilizing and destabilizing effects. The nature of the involvement of various tissue and membrane electrical properties is therefore complex, affecting several factors relevant to meandering at once. (c) 2002 American Institute of Physics. PMID- 12779612 TI - Enhanced self-termination of re-entrant arrhythmias as a pharmacological strategy for antiarrhythmic action. AB - Ventricular tachycardia and fibrillation are potentially lethal cardiac arrhythmias generated by high frequency, irregular spatio-temporal electrical activity. Re-entrant propagation has been demonstrated as a mechanism generating these arrhythmias in computational and in vitro animal models of these arrhythmias. Re-entry can be idealised in homogenous isotropic virtual cardiac tissues as spiral and scroll wave solutions of reaction-diffusion equations. A spiral wave in a bounded medium can be terminated if its core reaches a boundary. Ventricular tachyarrhythmias in patients are sometimes observed to spontaneously self-terminate. One possible mechanism for self-termination of a spiral wave is meander of its core to an inexcitable boundary. We have previously proposed the hypothesis that the spatial extent of meander of a re-entrant wave in the heart can be directly related to its probability of self-termination, and so inversely related to its lethality. Meander in two-dimensional virtual ventricular tissues based on the Oxsoft family of cell models, with membrane excitation parameters simulating the inherited long Q-T syndromes has been shown to be consistent with this hypothesis: the largest meander is seen in the syndrome with the lowest probability of death per arrhythmic episode. Here we extend our previous results to virtual tissues based on the Luo-Rudy family of models. Consistent with our hypothesis, for both families of models, whose different ionic mechanisms produce different patterns of meander, the LQT virtual tissue with the larger meander simulates the syndrome with the lower probability of death per episode. Further, we search the parameter space of the repolarizing currents to find their conductance parameter values that give increased meander of spiral waves. These parameters may provide targets for antiarrhythmic drugs designed to act by increasing the likelihood of self-termination of re-entrant arrhythmias. (c) 2002 American Institute of Physics. PMID- 12779613 TI - Multiple mechanisms of spiral wave breakup in a model of cardiac electrical activity. AB - It has become widely accepted that the most dangerous cardiac arrhythmias are due to reentrant waves, i.e., electrical wave(s) that recirculate repeatedly throughout the tissue at a higher frequency than the waves produced by the heart's natural pacemaker (sinoatrial node). However, the complicated structure of cardiac tissue, as well as the complex ionic currents in the cell, have made it extremely difficult to pinpoint the detailed dynamics of these life threatening reentrant arrhythmias. A simplified ionic model of the cardiac action potential (AP), which can be fitted to a wide variety of experimentally and numerically obtained mesoscopic characteristics of cardiac tissue such as AP shape and restitution of AP duration and conduction velocity, is used to explain many different mechanisms of spiral wave breakup which in principle can occur in cardiac tissue. Some, but not all, of these mechanisms have been observed before using other models; therefore, the purpose of this paper is to demonstrate them using just one framework model and to explain the different parameter regimes or physiological properties necessary for each mechanism (such as high or low excitability, corresponding to normal or ischemic tissue, spiral tip trajectory types, and tissue structures such as rotational anisotropy and periodic boundary conditions). Each mechanism is compared with data from other ionic models or experiments to illustrate that they are not model-specific phenomena. Movies showing all the breakup mechanisms are available at http://arrhythmia.hofstra.edu/breakup and at ftp://ftp.aip.org/epaps/chaos/E CHAOEH-12-039203/ INDEX.html. The fact that many different breakup mechanisms exist has important implications for antiarrhythmic drug design and for comparisons of fibrillation experiments using different species, electromechanical uncoupling drugs, and initiation protocols. (c) 2002 American Institute of Physics. PMID- 12779614 TI - Critical role of inhomogeneities in pacing termination of cardiac reentry. AB - Reentry around nonconducting ventricular scar tissue, a cause of lethal arrhythmias, is typically treated by rapid electrical stimulation from an implantable cardioverter defibrillator. However, the dynamical mechanisms of termination (success and failure) are poorly understood. To elucidate such mechanisms, we study the dynamics of pacing in one- and two-dimensional models of anatomical reentry. In a crucial realistic difference from previous studies of such systems, we have placed the pacing site away from the reentry circuit. Our model-independent results suggest that with such off-circuit pacing, the existence of inhomogeneity in the reentry circuit is essential for successful termination of tachycardia under certain conditions. Considering the critical role of such inhomogeneities may lead to more effective pacing algorithms. (c) 2002 American Institute of Physics. PMID- 12779615 TI - Resetting and annihilation of reentrant activity in a model of a one-dimensional loop of ventricular tissue. AB - Resetting and annihilation of reentrant activity by a single stimulus pulse (S1) or a pair (S1-S2) of coupled pulses are studied in a model of one-dimensional loop of cardiac tissue using a Beeler-Reuter-type ionic model. Different modes of reentry termination are described. The classical mode of termination by unidirectional block, in which a stimulus produces only a retrograde front that collides with the activation front of the reentry, can be obtained for both S1 and S1-S2 applied over a small vulnerable window. We demonstrate that another scenario of termination-that we term collision block-can also be induced by the S1-S2 protocol. This scenario is obtained over a much wider range of S1-S2 coupling intervals than the one leading to a unidirectional block. In the collision block, S1 produces a retrograde front, colliding with the activation front of the pre-existing reentry, and an antegrade front propagating in the same direction as the initial reentry. Then, S2 also produces an antegrade and a retrograde front. However, the propagation of these fronts in the spatial profile of repolarization left by S1 leads to a termination of the reentrant activity. More complex behaviors also occur in which the antegrade fronts produced by S1 and S2 both persist for several turns, displaying a growing alternation in action potential duration ("alternans amplification") that may lead to the termination of the reentrant activity. The hypothesis that both collision block and alternans amplification depend on the interaction between the action potential duration restitution curve and the recovery curve of conduction velocity is supported by the fact that the dynamical behaviors were reproduced using an integro-delay equation based on these two properties. We thus describe two new mechanisms (collision block and alternans amplification) whereby electrical stimulation can terminate reentrant activity. (c) 2002 American Institute of Physics. PMID- 12779616 TI - Spatiotemporal control of cardiac alternans. AB - Electrical alternans are believed to be linked to the onset of life-threatening ventricular arrhythmias and sudden cardiac death. Recent studies have shown that alternans can be suppressed temporally by dynamic feedback control of the pacing interval. Here we investigate theoretically whether control can suppress alternans both temporally and spatially in homogeneous tissue paced at a single site. We first carry out ionic model simulations in a one-dimensional cable geometry which show that control is only effective up to a maximum cable length that decreases sharply away from the alternans bifurcation point. We then explain this finding by a linear stability analysis of an amplitude equation that describes the spatiotemporal evolution of alternans. This analysis reveals that control failure above a critical cable length is caused by the formation of standing wave patterns of alternans that are eigenfunctions of a forced Helmholtz equation, and therefore remarkably analogous to sound harmonics in an open pipe. We discuss the implications of these results for using control to suppress alternans in the human ventricles as well as to probe fundamental aspects of alternans morphogenesis. (c) 2002 American Institute of Physics. PMID- 12779617 TI - Suppressing arrhythmias in cardiac models using overdrive pacing and calcium channel blockers. AB - Recent findings indicate that ventricular fibrillation might arise from spiral wave chaos. Our objective in this computational study was to investigate wave interactions in excitable media and to explore the feasibility of using overdrive pacing to suppress spiral wave chaos. This work is based on the finding that in excitable media, propagating waves with the highest excitation frequency eventually overtake all other waves. We analyzed the effects of low-amplitude, high-frequency pacing in one-dimensional and two-dimensional networks of coupled, excitable cells governed by the Luo-Rudy model. In the one-dimensional cardiac model, we found narrow high-frequency regions of 1:1 synchronization between the input stimulus and the system's response. The frequencies in this region were higher than the intrinsic spiral wave frequency of cardiac tissue. When we paced the two-dimensional cardiac model with frequencies from this region, we found that spiral wave chaos could, in some cases, be suppressed. When we coupled the overdrive pacing with calcium channel blockers, we found that spiral wave chaos could be suppressed in all cases. These findings suggest that low-amplitude, high frequency overdrive pacing, in combination with calcium channel inhibitors (e.g., class II or class IV antiarrhythmic drugs), may be useful for eliminating fibrillation. (c) 2002 American Institute of Physics. PMID- 12779618 TI - Termination of spiral wave breakup in a Fitzhugh-Nagumo model via short and long duration stimuli. AB - Rotating spiral waves have been observed in a variety of nonlinear biological and physical systems. Spiral waves are found in excitable and oscillatory systems and can be stationary, meander, or even degenerate into multiple unstable rotating waves (a process called "spiral wave breakup"). In the heart, spiral wave breakup is thought to be the underlying mechanism of cardiac fibrillation. The spatiotemporal complexity of multiple unstable spiral waves is difficult to control or terminate. Here, the mechanisms of the termination of spiral wave breakup in response to global stimulation are investigated. A modified Fitzhugh Nagumo model was used to represent cellular kinetics to study the role of the fast (activation) and slow (recovery) variables. This simplified model allows a theoretical analysis of the termination of spiral wave breakup via both short and long duration pulses. Simulations were carried out in both two-dimensional sheets and in a three-dimensional geometry of the heart ventricles. The short duration pulses affected only the fast variable and acted to reset wave propagation. Monophasic pulses excited tissue ahead of the wave front thus reducing the amount of excitable tissue. Biphasic shocks did the same, but they also acted to generate new wave fronts from the pre-existing wave tails by making some active regions excitable. Thus, if the short duration stimuli were strong enough, they acted to fill in excitable tissue via propagating wave fronts and terminated all activity. The long duration wave forms were selected such that they had a frequency spectrum similar to that of the pseudoelectrocardiograms recorded during fibrillation. These long duration wave forms affected both the recovery and activation variables, and the mechanism of unstable multiple spiral wave termination was different compared to the short duration wave forms. If the long duration stimuli were strong enough, they acted to alter the "state" (i.e., combination of fast and slow variables) of the tissue throughout 1.5 cycles, thus "conditioning" the tissue such that by the end of the stimuli almost no excitable tissue remained. The peak current, total energy, and average power of stimuli required to terminate spiral wave breakup were less for the long duration wave forms compared to the short duration wave forms. In addition, closed loop feedback via stimulation with a wave form that was the difference of the pseudoelectrocardiogram and a strongly periodic chaotic signal was successful at terminating spiral wave breakup. These results suggest that it may be possible to improve cardiac defibrillation efficacy by using long duration wave forms to affect recovery variables in the heart as opposed to the traditional brief duration wave forms that act only on the fast variables. (c) 2002 American Institute of Physics. PMID- 12779619 TI - Progress toward controlling in vivo fibrillating sheep atria using a nonlinear dynamics-based closed-loop feedback method. AB - We describe preliminary experiments on controlling in vivo atrial fibrillation using a closed-loop feedback protocol that measures the dynamics of the right atrium at a single spatial location and applies control perturbations at a single spatial location. This study allows investigation of control of cardiac dynamics in a preparation that is physiologically close to an in vivo human heart. The spatial-temporal response of the fibrillating sheep atrium is measured using a multi-channel electronic recording system to assess the control effectiveness. In an attempt to suppress fibrillation, we implement a scheme that paces occasionally the cardiac muscle with small shocks. When successful, the inter activation time interval is the same and electrical stimuli are only applied when the controller senses that the dynamics are beginning to depart from the desired periodic rhythm. The shock timing is adjusted in real time using a control algorithm that attempts to synchronize the most recently measured inter activation interval with the previous interval by inducing an activation at a time projected by the algorithm. The scheme is "single-sided" in that it can only shorten the inter-activation time but not lengthen it. Using probability distributions of the inter-activation time intervals, we find that the feedback protocol is not effective in regularizing the dynamics. One possible reason for the less-than-successful results is that the controller often attempts to stimulate the tissue while it is still in the refractory state and hence it does not induce an activation. (c) 2002 American Institute of Physics. PMID- 12779620 TI - Shock-induced arrhythmogenesis in the myocardium. AB - The focus of this article is the investigation of the electrical behavior of the normal myocardium following the delivery of high-strength defibrillation shocks. To achieve its goal, the study employs a complex three-dimensional defibrillation model of a slice of the canine heart characterized with realistic geometry and fiber architecture. Defibrillation shocks of various strengths and electrode configurations are delivered to the model preparation in which a sustained ventricular tachycardia is induced. Instead of analyzing the post-shock electrical events as progressions of transmembrane potential maps, the study examines the evolution of the postshock phase singularities (PSs) which represent the organizing centers of reentry. The simulation results demonstrate that the shock induces numerous PSs the majority of which vanish before the reentrant wavefronts associated with them complete half of a single rotation. Failed shocks are characterized with one or more PSs that survive the initial period of PS annihilation to establish a new postshock arrhythmia. The increase in shock strength results in an overall decrease of the number of PSs that survive over 200 ms after the end of the shock; however, the exact behavior of the PSs is strongly dependent on the shock electrode configuration. (c) 2002 American Institute of Physics. PMID- 12779621 TI - Artifacts, assumptions, and ambiguity: Pitfalls in comparing experimental results to numerical simulations when studying electrical stimulation of the heart. AB - Insidious experimental artifacts and invalid theoretical assumptions complicate the comparison of numerical predictions and observed data. Such difficulties are particularly troublesome when studying electrical stimulation of the heart. During unipolar stimulation of cardiac tissue, the artifacts include nonlinearity of membrane dyes, optical signals blocked by the stimulating electrode, averaging of optical signals with depth, lateral averaging of optical signals, limitations of the current source, and the use of excitation-contraction uncouplers. The assumptions involve electroporation, membrane models, electrode size, the perfusing bath, incorrect model parameters, the applicability of a continuum model, and tissue damage. Comparisons of theory and experiment during far-field stimulation are limited by many of these same factors, plus artifacts from plunge and epicardial recording electrodes and assumptions about the fiber angle at an insulating boundary. These pitfalls must be overcome in order to understand quantitatively how the heart responds to an electrical stimulus. (c) 2002 American Institute of Physics. PMID- 12779622 TI - Critical points and transitions in an electric power transmission model for cascading failure blackouts. AB - Cascading failures in large-scale electric power transmission systems are an important cause of blackouts. Analysis of North American blackout data has revealed power law (algebraic) tails in the blackout size probability distribution which suggests a dynamical origin. With this observation as motivation, we examine cascading failure in a simplified transmission system model as load power demand is increased. The model represents generators, loads, the transmission line network, and the operating limits on these components. Two types of critical points are identified and are characterized by transmission line flow limits and generator capability limits, respectively. Results are obtained for tree networks of a regular form and a more realistic 118-node network. It is found that operation near critical points can produce power law tails in the blackout size probability distribution similar to those observed. The complex nature of the solution space due to the interaction of the two critical points is examined.(c) 2002 American Institute of Physics. PMID- 12779623 TI - Targeting in dissipative chaotic systems: A survey. AB - The large number of unstable equilibrium modes embedded in the strange attractor of dissipative chaotic systems usually presents a sufficiently rich repertoire for the choice of the desirable motion as a target. Once the system is close enough to the chosen target local stabilization techniques can be employed to capture the system within the desired motion. The ergodic behavior of chaotic systems on their strange attractors guarantees that the system will eventually visit a close neighborhood of the target. However, for arbitrary initial conditions within the basin of attraction of the strange attractor the waiting time for such a visit may be intolerably long. In order to reduce the long waiting time it usually becomes indispensable to employ an appropriate method of targeting, which refers to the task of steering the system toward the close neighborhood of the target. This paper provides a survey of targeting methods proposed in the literature for dissipative chaotic systems. (c) 2002 American Institute of Physics. PMID- 12779624 TI - Multifractal chaotic attractors in a system of delay-differential equations modeling road traffic. AB - We study a system of delay-differential equations modeling single-lane road traffic. The cars move in a closed circuit and the system's variables are each car's velocity and the distance to the car ahead. For low and high values of traffic density the system has a stable equilibrium solution, corresponding to the uniform flow. Gradually decreasing the density from high to intermediate values we observe a sequence of supercritical Hopf bifurcations forming multistable limit cycles, corresponding to flow regimes with periodically moving traffic jams. Using an asymptotic technique we find approximately small limit cycles born at Hopf bifurcations and numerically preform their global continuations with decreasing density. For sufficiently large delay the system passes to chaos following the Ruelle-Takens-Newhouse scenario (limit cycles-two tori-three-tori-chaotic attractors). We find that chaotic and nonchaotic attractors coexist for the same parameter values and that chaotic attractors have a broad multifractal spectrum. (c) 2002 American Institute of Physics. PMID- 12779625 TI - Generation of undular bores in the shelves of slowly-varying solitary waves. AB - We study the long-time evolution of the trailing shelves that form behind solitary waves moving through an inhomogeneous medium, within the framework of the variable-coefficient Korteweg-de Vries equation. We show that the nonlinear evolution of the shelf leads typically to the generation of an undular bore and an expansion fan, which form apart but start to overlap and nonlinearly interact after a certain time interval. The interaction zone expands with time and asymptotically as time goes to infinity occupies the whole perturbed region. Its oscillatory structure strongly depends on the sign of the inhomogeneity gradient of the variable background medium. We describe the nonlinear evolution of the shelves in terms of exact solutions to the KdV-Whitham equations with natural boundary conditions for the Riemann invariants. These analytic solutions, in particular, describe the generation of small "secondary" solitary waves in the trailing shelves, a process observed earlier in various numerical simulations. (c) 2002 American Institute of Physics. PMID- 12779626 TI - An extension to chaos control via Lie derivatives: Fully linearizable systems. AB - The technique of using Lie derivatives to control chaos introduced by Kocarev et al. [Chaos, Solitons Fractals 9, 1359-1366 (1998)] is extended in this contribution. Here, by using Lie derivatives in an extended space state, it is proved that chaos can be practically suppressed via feedback in spite of the Lie derivative being ill-posed at the reference. The main idea is to construct a dynamically equivalent system. In this way, the chaotic system can be practically stabilized around any point of singularity x(0). The Lorenz equation is used as an illustrative example to show the application in the chaos control context. (c) 2002 American Institute of Physics. PMID- 12779627 TI - Analysis of the Fenton-Karma model through an approximation by a one-dimensional map. AB - The Fenton-Karma model is a simplification of complex ionic models of cardiac membrane that reproduces quantitatively many of the characteristics of heart cells; its behavior is simple enough to be understood analytically. In this paper, a map is derived that approximates the response of the Fenton-Karma model to stimulation in zero spatial dimensions. This map contains some amount of memory, describing the action potential duration as a function of the previous diastolic interval and the previous action potential duration. Results obtained from iteration of the map and numerical simulations of the Fenton-Karma model are in good agreement. In particular, the iterated map admits different types of solutions corresponding to various dynamical behavior of the cardiac cell, such as 1:1 and 2:1 patterns. (c) 2002 American Institute of Physics. PMID- 12779628 TI - Transport in a slowly perturbed convective cell flow. AB - We study transport properties in a simple model of two-dimensional roll convection under a slow periodic (period of order 1/ varepsilon >>1) perturbation. The problem is considered in terms of conservation of the adiabatic invariant. It is shown that the adiabatic invariant is well conserved in the system. It results in almost regular dynamics on large time scales (of order approximately varepsilon (-3) ln varepsilon ) and hence, fast transport. We study both generic systems and an example having some symmetry. (c) 2002 American Institute of Physics. PMID- 12779629 TI - Border collision bifurcations at the change of state-space dimension. AB - We present the theory of border collision bifurcation for the special case where the state space is piecewise smooth, but two-dimensional in one side of the borderline, and one dimensional in the other side. This situation occurs in a class of switching circuits widely used in power electronic industry. We analyze this particular class of bifurcations in terms of the normal form, where the determinant of the Jacobian matrix at one side of the borderline is greater than unity in magnitude, and in the other side it is zero. (c) 2002 American Institute of Physics. PMID- 12779630 TI - Generation of large-amplitude solitons in the extended Korteweg-de Vries equation. AB - We study the extended Korteweg-de Vries equation, that is, the usual Korteweg-de Vries equation but with the inclusion of an extra cubic nonlinear term, for the case when the coefficient of the cubic nonlinear term has an opposite polarity to that of the coefficient of the linear dispersive term. As this equation is integrable, the number and type of solitons formed can be determined from an appropriate spectral problem. For initial disturbances of small amplitude, the number and type of solitons generated is similar to the well-known situation for the Korteweg-de Vries equation. However, our interest here is in initial disturbances of larger amplitude, for which there is the possibility of the generation of large-amplitude "table-top" solitons as well as small-amplitude solitons similar to the solitons of the Korteweg-de Vries equation. For this case, and in contrast to some earlier results which assumed that an initial disturbance in the shape of a rectangular box would be typical, we show that the number and type of solitons formed depend crucially on the disturbance shape, and change drastically when the initial disturbance is changed from a rectangular box to a "sech"-profile. (c) 2002 American Institute of Physics. PMID- 12779631 TI - Self-similarity and renormalization in chaotic dynamics. PMID- 12779632 TI - Stable periodic motions in the problem on passage through a separatrix. AB - A Hamiltonian system with one degree of freedom depending on a slowly periodically varying in time parameter is considered. For every fixed value of the parameter there are separatrices on the phase portrait of the system. When parameter is changing in time, these separatrices are pulsing slowly periodically, and phase points of the system cross them repeatedly. In numeric experiments region swept by pulsing separatrices looks like a region of chaotic motion. However, it is shown in the present paper that if the system possesses some additional symmetry (like a pendulum in a slowly varying gravitational field), then typically in the region in question there are many periodic solutions surrounded by stability islands; total measure of these islands does not vanish and does not tend to 0 as rate of changing of the parameter tends to 0.(c) 1997 American Institute of Physics. PMID- 12779633 TI - Pesin's dimension for Poincare recurrences. AB - A new characteristic of Poincare recurrences is introduced. It describes an average return time in the framework of a general construction for dimension-like characteristics. Some examples are considered including rotations on the circle and the Denjoy example. (c) 1997 American Institute of Physics. PMID- 12779634 TI - Dynamics of spatial averages. AB - We study the dynamics of spatial averages of spatially extended dynamical systems. We present various examples of lattice dynamical systems to show the possibility of different behaviors, including asymptotically constant, periodic, and non-periodic, of spatial averages. We explain that the fluctuation in spatial averages is caused by the transitivity and the lack of symmetry of the dynamics of local subsystems. (c) 1997 American Institute of Physics. PMID- 12779635 TI - On a general concept of multifractality: Multifractal spectra for dimensions, entropies, and Lyapunov exponents. Multifractal rigidity. AB - We introduce the mathematical concept of multifractality and describe various multifractal spectra for dynamical systems, including spectra for dimensions and spectra for entropies. We support the study by providing some physical motivation and describing several nontrivial examples. Among them are subshifts of finite type and one-dimensional Markov maps. An essential part of the article is devoted to the concept of multifractal rigidity. In particular, we use the multifractal spectra to obtain a "physical" classification of dynamical systems. For a class of Markov maps, we show that, if the multifractal spectra for dimensions of two maps coincide, then the maps are differentiably equivalent. (c) 1997 American Institute of Physics. PMID- 12779636 TI - Monte Carlo methods for turbulent tracers with long range and fractal random velocity fields. AB - Monte Carlo methods for computing various statistical aspects of turbulent diffusion with long range correlated and even fractal random velocity fields are described here. A simple explicit exactly solvable model with complex regimes of scaling behavior including trapping, subdiffusion, and superdiffusion is utilized to compare and contrast the capabilities of conventional Monte Carlo procedures such as the Fourier method and the moving average method; explicit numerical examples are presented which demonstrate the poor convergence of these conventional methods in various regimes with long range velocity correlations. A new method for computing fractal random fields involving wavelets and random plane waves developed recently by two of the authors [J. Comput. Phys. 117, 146 (1995)] is applied to compute pair dispersion over many decades for systematic families of anisotropic fractal velocity fields with the Kolmogorov spectrum. The important associated preconstant for pair dispersion in the Richardson law in these anisotropic settings is compared with the one obtained over many decades recently by two of the authors [Phys. Fluids 8, 1052 (1996)] for an isotropic fractal field with the Kolmogorov spectrum. (c) 1997 American Institute of Physics. PMID- 12779637 TI - Quasi-periodicity, global stability and scaling in a model of Hamiltonian round off. AB - We investigate the effects of round-off errors on the orbits of a linear symplectic map of the plane, with rational rotation number nu=p/q. Uniform discretization transforms this map into a permutation of the integer lattice Z(2). We study in detail the case q=5, exploiting the correspondence between Z and a suitable domain of algebraic integers. We completely classify the orbits, proving that all of them are periodic. Using higher-dimensional embedding, we establish the quasi-periodicity of the phase portrait. We show that the model exhibits asymptotic scaling of the periodic orbits and a long-range clustering property similar to that found in repetitive tilings of the plane. (c) 1997 American Institute of Physics. PMID- 12779638 TI - Entropy conservation as h(T(&mgr;) ) approximately lambda(&mgr;) (+)d(&mgr;) in neurobiological dynamical systems. AB - That the topological entropy, h(T(&mgr;) ), of a C(1M, of a surface, M, upon which invariant measure(s) &mgr; are concentrated, varies as the product of its average leading Lyapunov characteristic exponent, lambda(&mgr;), and the Hausdorff dimension of its support, d(&mgr;),was proven by Pesin [Russ. Math Surveys 32, 55-114 (1977)] for nonuniform partial hyperbolic systems and by Ledreppier and Young [Ergod. Theor. Dyn. Syst. 2, 109-123 (1982)], and Manning [Ergod. Theor. Dyn. Syst. 1, 451-459 (1981)] for uniformly hyperbolic (Axiom A) diffeomorphisms. When considered in conjunction with the post-Shannon information encoding theorems of Adler [Trans. Am. Math. Soc. 114, 309-319 (1965); Mem. Am. Math. Soc., No. 219 (1979)] and others, this suggests a way to differentiate equal entropy behaviors in systems with varying patterns of dynamical behaviors. Here we show this relation to be useful in the quantitative discrimination among the behaviors of abstract neuronal models and two real, finite time, partially and nonuniformly hyperbolic, brain-related dynamical systems. We observe a trade-off in finite time between two competing dynamical processes, jittery sticking (tending to increase d(&mgr;)) and convective escaping (more prominently incrementing lambda(&mgr;) (+)). In finite time systems, these changes in combination can statistically conserve the dynamical entropy, h(T(&mgr;) ), while altering the Levy characteristic exponent, alpha (describing the tail of the density distribution of observables, rho(x) approximately exp-gammamid R:xmid R:(alpha),10.5 implicates sequential correlations and H(*)<0.5 sequential anticorrelation. When the relation h(T(&mgr;) )=lambda(&mgr;) (+)d&mgr; fails, the way it does so provides information about the system. (c) 1997 American Institute of Physics. PMID- 12779639 TI - Cosmic lacunarity. AB - The present distribution of galaxies in space is a remnant of their formation and interaction. On a large enough scale, we may represent the galaxies as a set of points and quantify the structures in this set by its generalized dimensions [Beck and Schlogl, Thermodynamics of Chaotic Systems (Cambridge University Press, Cambridge, 1986); Paladin and Vulpiani, Phys. Rep. 156, 147 (1987)]. The results of such evaluation are often taken to be evidence of a fractal (or multifractal) distribution of galaxies. However, those results, for some scales, may also reveal the presence of singularities formed in the gravitational processes that produce structure in the galaxy distribution. To try to make some decision about this issue, we look for the more subtle galactic lacunarity. We believe that this quantity is discernible in the currently available data and that it provides important evidence on the galaxy formation process. (c) 1997 American Institute of Physics. PMID- 12779640 TI - The multifractal analysis of Gibbs measures: Motivation, mathematical foundation, and examples. AB - We first motivate the study of multifractals. We then present a rigorous mathematical foundation for the multifractal analysis of Gibbs measures invariant under dynamical systems. Finally we effect a complete multifractal analysis for several classes of hyperbolic dynamical systems. (c) 1997 American Institute of Physics. PMID- 12779641 TI - Plume dynamics in quasi-2D turbulent convection. AB - We have studied turbulent convection in a vertical thin (Hele-Shaw) cell at very high Rayleigh numbers (up to 7x10(4) times the value for convective onset) through experiment, simulation, and analysis. Experimentally, convection is driven by an imposed concentration gradient in an isothermal cell. Model equations treat the fields in two dimensions, with the reduced dimension exerting its influence through a linear wall friction. Linear stability analysis of these equations demonstrates that as the thickness of the cell tends to zero, the critical Rayleigh number and wave number for convective onset do not depend on the velocity conditions at the top and bottom boundaries (i.e., no-slip or stress free). At finite cell thickness delta, however, solutions with different boundary conditions behave differently. We simulate the model equations numerically for both types of boundary conditions. Time sequences of the full concentration fields from experiment and simulation display a large number of solutal plumes that are born in thin concentration boundary layers, merge to form vertical channels, and sometimes split at their tips via a Rayleigh-Taylor instability. Power spectra of the concentration field reveal scaling regions with slopes that depend on the Rayleigh number. We examine the scaling of nondimensional heat flux (the Nusselt number, Nu) and rms vertical velocity (the Peclet number, Pe) with the Rayleigh number (Ra(*)) for the simulations. Both no-slip and stress-free solutions exhibit the scaling NuRa(*) approximately Pe(2) that we develop from simple arguments involving dynamics in the interior, away from cell boundaries. In addition, for stress-free solutions a second relation, Nu approximately nPe, is dictated by stagnation-point flows occurring at the horizontal boundaries; n is the number of plumes per unit length. No-slip solutions exhibit no such organization of the boundary flow and the results appear to agree with Priestley's prediction of Nu approximately Ra(1/3). (c) 1997 American Institute of Physics. PMID- 12779642 TI - Indecomposable continua in dynamical systems with noise: Fluid flow past an array of cylinders. AB - Standard dynamical systems theory is based on the study of invariant sets. However, when noise is added, there are no bounded invariant sets. Our goal is then to study the fractal structure that exists even with noise. The problem we investigate is fluid flow past an array of cylinders. We study a parameter range for which there is a periodic oscillation of the fluid, represented by vortices being shed past each cylinder. Since the motion is periodic in time, we can study a time-1 Poincare map. Then we add a small amount of noise, so that on each iteration the Poincare map is perturbed smoothly, but differently for each time cycle. Fix an x coordinate x(0) and an initial time t(0). We discuss when the set of initial points at a time t(0) whose trajectory (x(t),y(t)) is semibounded (i.e., x(t)>x(0) for all time) has a fractal structure called an indecomposable continuum. We believe that the indecomposable continuum will become a fundamental object in the study of dynamical systems with noise. (c) 1997 American Institute of Physics. PMID- 12779643 TI - Average exit time for volume-preserving maps. AB - For a volume-preserving map, we show that the exit time averaged over the entry set of a region is given by the ratio of the measure of the accessible subset of the region to that of the entry set. This result is primarily of interest to show two things: First, it gives a simple bound on the algebraic decay exponent of the survival probability. Second, it gives a tool for computing the measure of the accessible set. We use this to compute the measure of the bounded orbits for the Henon quadratic map. (c) 1997 American Institute of Physics. PMID- 12779644 TI - Parabolic resonances and instabilities. AB - A parabolic resonance is formed when an integrable two-degrees-of-freedom (d.o.f.) Hamiltonian system possessing a circle of parabolic fixed points is perturbed. It is proved that its occurrence is generic for one parameter families (co-dimension one phenomenon) of near-integrable, two d.o.f. Hamiltonian systems. Numerical experiments indicate that the motion near a parabolic resonance exhibits a new type of chaotic behavior which includes instabilities in some directions and long trapping times in others. Moreover, in a degenerate case, near a flat parabolic resonance, large scale instabilities appear. A model arising from an atmospherical study is shown to exhibit flat parabolic resonance. This supplies a simple mechanism for the transport of particles with small (i.e. atmospherically relevant) initial velocities from the vicinity of the equator to high latitudes. A modification of the model which allows the development of atmospherical jets unfolds the degeneracy, yet traces of the flat instabilities are clearly observed. (c) 1997 American Institute of Physics. PMID- 12779645 TI - Self-similarity, renormalization, and phase space nonuniformity of Hamiltonian chaotic dynamics. AB - A detailed description of fractional kinetics is given in connection to islands' topology in the phase space of a system. The method of renormalization group is applied to the fractional kinetic equation in order to obtain characteristic exponents of the fractional space and time derivatives, and an analytic expression for the transport exponents. Numerous simulations for the web-map and standard map demonstrate different results of the theory. Special attention is applied to study the singular zone, a domain near the island boundary with a self similar hierarchy of subislands. The birth and collapse of islands of different types are considered. (c) 1997 American Institute of Physics. PMID- 12779646 TI - Chaotic transmission of waves and "cooling" of signals. AB - Ray dynamics in waveguide media exhibits chaotic motion. For a finite length of propagation, the large distance asymptotics is not uniform and represents a complicated combination of bunches of rays with different intermediate asymptotics. The origin of the phenomena that we call "chaotic transmission," lies in the nonuniformity of the phase space with sticky domains near the boundary of islands. We demonstrate different fractal properties of ray propagation using underwater acoustics as an example. The phenomenon of the kind of Levy flights can occur and it can be used as a mechanism of cooling of signals when the width of spatial spectra dispersion is significantly reduced. (c) 1997 American Institute of Physics. PMID- 12779647 TI - Dynamics of a pair of spherical gravitating shells. AB - The dynamical N body problem for a system of mass points interacting solely through gravitational forces is not integrable. The difficulties which arise in constructing accurate numerical codes for simulating the motion over long time scales are legend. Thus, in order to test their theories, astronomers and astrophysicists resort to simpler, one-dimensional models which avoid the problems of binary formation, escape, and the singularity of the inverse square force law. To date, the most frequently employed "test" model consists of a system of parallel mass sheets moving perpendicular to their surface. While this system avoids all of the above problems, the time scale for reaching equilibrium is extremely long and probably arises from the system's weak ergodic properties, which become manifest even in the three sheet system. Here we consider a different one-dimensional gravitating system consisting of nonrotating concentric mass shells. For the case of two shells we investigate the structure of the phase space by studying the stability of periodic trajectories. By employing an event driven algorithm, we are able to directly investigate the influence of the singularity without having to resort to regularization of the force. Although stable structures are present at every energy, we find that the ergodic properties of this system are more robust than its planar counterpart. (c) 1997 American Institute of Physics. PMID- 12779648 TI - On the chaotic nature of turbulence observed in benchmark analysis of nonlinear plasma simulation. AB - Simulational results of two dissipative interchange turbulence (Rayleigh-Taylor type instability with dissipation) models with the same physics are compared. The convective nonlinearity is the nonlinear mechanism in the models. They are shown to have different time evolutions in the nonlinear phase due to the different initial value which is attributed to the initial noise. In the first model (A), a single pressure representing the sum of the ion and electron components is used (one-fluid model). In the second model (B) the ion and electron components of the pressure fields are independently solved (two-fluid model). Both models become physically identical if we set ion and electron pressure fields to be equal in the model (B). The initial conditions only differ by the infinitesimally small initial noise due to the roundoff errors which comes from the finite difference but not the differentiation. This noise grows in accordance with the nonlinear development of the turbulence mode. Interaction with an intrinsic nonlinearity of the system makes the noise grow, whose contribution becomes almost the same magnitude of the fluctuation itself in the results. The instantaneous deviation shows the chaotic characteristics of the turbulence. (c) 1997 American Institute of Physics. PMID- 12779649 TI - Cycles homoclinic to chaotic sets; robustness and resonance. AB - For dynamical systems possessing invariant subspaces one can have a robust homoclinic cycle to a chaotic set. If such a cycle is stable, it manifests itself as long periods of quiescent chaotic behaviour interrupted by sudden transient 'bursts'. The time between the transients increases as the trajectory approaches the cycle. This behavior for a cycle connecting symmetrically related chaotic sets has been called 'cycling chaos' by Dellnitz et al. [IEEE Trans. Circ. Sys. I 42, 821-823 (1995)]. We characterise such cycles and their stability by means of normal Lyapunov exponents. We find persistence of states that are not Lyapunov stable but still attracting, and also states that are approximately periodic. For systems possessing a skew-product structure (such as naturally arises in chaotically forced systems) we show that the asymptotic stability and the attractivity of the cycle depends in a crucial way on what we call the footprint of the cycle. This is the spectrum of Lyapunov exponents of the chaotic invariant set in the expanding and contracting directions of the cycle. Numerical simulations and calculations for an example system of a homoclinic cycle parametrically forced by a Rossler attractor are presented; here we observe the creation of nearby chaotic attractors at resonance of transverse Lyapunov exponents. (c) 1997 American Institute of Physics. PMID- 12779650 TI - Exploring invariant sets and invariant measures. AB - We propose a method to explore invariant measures of dynamical systems. The method is based on numerical tools which directly compute invariant sets using a subdivision technique, and invariant measures by a discretization of the Frobenius-Perron operator. Appropriate visualization tools help to analyze the numerical results and to understand important aspects of the underlying dynamics. This will be illustrated for examples provided by the Lorenz system. (c) 1997 American Institute of Physics. PMID- 12779651 TI - Branching equation of Andronov-Hopf bifurcation under group symmetry conditions. AB - In branching theory of solutions of nonlinear equations group analysis methods [Ovsyannikov, Group Analysis of Differential Equations (Nauka, Moscow, 1978); Lectures on the Theory of Group Properties of Differential Equations (Novosibirsk University, Novosibirsk, 1966)] give the general approach for the construction of the complete form of branching equation and its subsequent investigation. These methods are applied here to the general situation of Andronov-Hopf bifurcation when there are some multiple semisimple eigenvalues on imaginary axis. (c) 1997 American Institute of Physics. PMID- 12779652 TI - Turning point properties as a method for the characterization of the ergodic dynamics of one-dimensional iterative maps. AB - Dynamical as well as statistical properties of the ergodic and fully developed chaotic dynamics of iterative maps are investigated by means of a turning point analysis. The turning points of a trajectory are hereby defined as the local maxima and minima of the trajectory. An examination of the turning point density directly provides us with the information of the position of the fixed point for the corresponding dynamical system. Dividing the ergodic dynamics into phases consisting of turning points and nonturning points, respectively, elucidates the understanding of the organization of the chaotic dynamics for maps. The turning point map contains information on any iteration of the dynamical law and is shown to possess an asymptotic scaling behaviour which is responsible for the assignment of dynamical structures to the environment of the two fixed points of the map. Universal statistical turning point properties are derived for doubly symmetric maps. Possible applications of the observed turning point properties for the analysis of time series are discussed in some detail. (c) 1997 American Institute of Physics. PMID- 12779653 TI - Separatrix reconnection and periodic orbit annihilation in the Harper map. AB - Structure of the periodic accelerator orbits of the Harper map is investigated in detail from the viewpoint of underlying scenario of chaos in the area preserving nontwist map. Since the twist function of the Harper map is free from the polynomial local approximation, it admits rigorous treatment for the entire range of phase variable. The results obtained in the present analysis describes generic novel phenomena, which are outside of the applicability of the Kolmogorov-Arnol'd Moser theory. (c) 1997 American Institute of Physics. PMID- 12779654 TI - Construction of the Jordan basis for the Baker map. AB - The Jordan canonical form basis states for an invertible chaotic map, the Baker map, are constructed. A straightforwardly obtained recursion formula is presented for construction of the Jordan states and of the spectral decomposition of the Frobenius-Perron evolution operator. Comparison of this method with earlier, subdynamics techniques demonstrates that it is much more direct and simpler. The physical significance of the Jordan states is approached from the point of view of an entropy evolution equation. The method is also applied to the Bernoulli map, yielding its eigenstates more straightforwardly than done previously. (c) 1997 American Institute of Physics. PMID- 12779655 TI - The interaction of resonances in a weakly nonlinear oscillator with a quasiperiodic excitation. AB - Conditions for chaos onset due to the interaction of resonances in a weakly nonlinear oscillator with a quasiperiodic excitation are investigated. The focus is made on the interaction of the 1:1 resonance with that of 1:m, where m is an arbitrary integer number. Analytical conditions of the chaos arising are obtained through the combined application of the averaging procedure and Melnikov's technique. The comparison study of these conditions with those following from standard Chirikov's approach is provided. (c) 1997 American Institute of Physics. PMID- 12779656 TI - Parameter renormalization of maps based on potential function. AB - A systematic way for deriving the parameter renormalization group equation for one-dimensional maps is presented and the critical behavior of periodic doubling is investigated. Introducing a formal potential function in one-parameter cases, it is shown that accumulation points correspond to local potential maxima and universal constants are easily determined. The estimates of accumulation points and universal constants match the known values asymptotically when the order of potential grows large. The potential function shows scaling in the parameter space with the universal convergent rate at the accumulation point similar to the Feigenbaum universal function. For two-parameter cases, a parameter reduction transformation is found to be useful to determine some important fixed points. A locally defined potential function is introduced and its scaling property is discussed. (c) 1997 American Institute of Physics. PMID- 12779657 TI - Control of noisy chaotic motion in a system with nonlinear excitation and restoring forces. AB - In this study we examine the complex and chaotic oscillations of a dynamical system with nonlinear excitation and restoring forces for the purpose of controlling these oscillatory states. The physical system, modeled as a system of first-order nonlinear ordinary differential equations, takes into account a geometric nonlinearity in the restoring force, a quadratic viscous drag, and a harmonic excitation force. It is controlled using small perturbations about a selected unstable cycle and control is instigated for periodic cycles of varying periodicities. The controller, when applied on the dynamical system with additive random noise in the excitation, successfully controls the system with noise levels in excess of 5% of the total energy, giving the first evidence that (stochastic) control of these systems is possible. (c) 1997 American Institute of Physics. PMID- 12779658 TI - On the global orbits in a bistable CML. AB - In an infinite one-dimensional coupled map lattice (CML) for which the local map is piecewise affine and bistable, we study the global orbits using a spatiotemporal coding introduced in a previous work. The set of all the fixed points is first considered. It is shown that, under some restrictions on the parameters, the latter is a Cantor set, and we introduce an order to study the fixed points' existence. This also involves the proof of the coexistence of propagating fronts and stationary structures. In the second part, we analyze the global orbits which occur for strong coupling using the splitting of the dynamics into two independent (sub-)lattices, and emphasize the description of various traveling structures. (c) 1997 American Institute of Physics. PMID- 12779659 TI - Dynamics of coupled maps with a conservation law. AB - A particularly simple model belonging to a wide class of coupled maps which obey a local conservation law is studied. The phase structure of the system and the types of the phase transitions are determined. It is argued that the structure of the phase diagram is robust with respect to mild violations of the conservation law. Critical exponents possibly determining a new universality class are calculated for a set of independent order parameters. Numerical evidence is produced suggesting that the singularity in the density of Lyapunov exponents at lambda=0 is a reflection of the singularity in the density of Fourier modes (a "Van Hove" singularity) and disappears if the conservation law is broken. Applicability of the Lyapunov dimension to the description of spatiotemporal chaos in a system with a conservation law is discussed. (c) 1997 American Institute of Physics. PMID- 12779660 TI - Numerical modeling of self-propagating polymerization fronts: The role of kinetics on front stability. AB - Frontal propagation of a highly exothermic polymerization reaction in a liquid is studied with the goal of developing a mathematical model of the process. As a model case we consider monomers such as methacrylic acid and n-butyl acrylate with peroxide initiators, although the model is not limited to these reactants and can be applied to any system with the similar basic polymerization mechanism. A three-step reaction mechanism, including initiation, propagation and termination steps, as well as a more simple one-step mechanism, were considered. For the one-step mechanism the loss of stability of propagating front was observed as a sequence of period doubling bifurcations of the front velocity. It was shown that the one-step model cannot account for less than 100% conversion and product inhomogeneities as a result of front instability, therefore the three step mechanism was exploited. The phenomenon of superadiabatic combustion temperature was observed beyond the Hopf bifurcation point for both kinetic schemes and supported by the experimental measurements. One- and two-dimensional numerical simulations were performed to observe various planar and nonplanar periodic modes, and the results for different kinetic schemes were compared. It was found that stability of the frontal mode for a one-step reaction mechanism does not differ for 1-D and 2-D cases. For the three-step reaction mechanism 2-D solutions turned out to be more stable with respect to the appearance of nonplanar periodic modes than corresponding 1-D solutions. Higher Zeldovich numbers (i.e., higher effective activation energies or lower initial temperatures) are necessary for the existence of planar and nonplanar periodic modes in the 2-D reactor with walls than in the 1-D case. (c) 1997 American Institute of Physics. PMID- 12779661 TI - Irregular phase slipping in a model for a laser with injected signal. AB - In a model for a single mode laser with injected signal we find irregular pulsing of the output signal. During the turbulent time intervals the relative phase between the internal and the external field slips by an integer multiple of 2pi, where the multiplicity varies irregularly from pulse to pulse. Between the pulses, in the laminar time intervals, the relative phase is locked. We compare this behavior with the results of various other laser models. By the investigation of this interesting mechanism for the creation of phase chaos in a driven oscillator, we learn which properties are responsible for the irregular interruption of the phase locking. (c) 1997 American Institute of Physics. PMID- 12779662 TI - Modelling complex transient oscillations for the BZ reaction in a batch reactor. AB - The recent observations of transient complex oscillations in the Belousov Zhabotinsky (BZ) reaction in a batch reactor are confirmed and an attempt is made to model this behavior using the Gyorgyi-Field mechanism (an extension of the classic Field-Koros-Noyes model). It is seen that the concentration of bromomalonic acid plays an important role, acting somewhat like a "slowly-varying parameter," causing the system to sweep through a region of oscillatory response even in the absence of reactant consumption. Complex behavior is not observed in the model if the usual "pre-equilibrium" assumption in made for the intermediate species BrO(2) but does arise if a full steady-state approximation is applied to this radical. These results may bring into question some assumptions made in the usual treatment of the cerium-catalyzed BZ system in closed reactors. (c) 1997 American Institute of Physics. PMID- 12779663 TI - The convergence of chaotic integrals. AB - We review the convergence of chaotic integrals computed by Monte Carlo simulation, the trace method, dynamical zeta function, and Fredholm determinant on a simple one-dimensional example: the parabola repeller. There is a dramatic difference in convergence between these approaches. The convergence of the Monte Carlo method follows an inverse power law, whereas the trace method and dynamical zeta function converge exponentially, and the Fredholm determinant converges faster than any exponential. (c) 1997 American Institute of Physics. PMID- 12779664 TI - Nonintegrability of two problems in vortex dynamics. AB - An analysis is presented of two problems in vortex dynamics whose equations can be written in the Hamiltonian form. They are: the interaction of three coaxial vortex rings and the motion of four point vortices on a sphere. The nonintegrability of these problems in the restricted formulation is demonstrated analytically by the method of split separatrices, using a small parameter. (c) 1997 American Institute of Physics. PMID- 12779665 TI - Bifurcation of the branching of a cycle in n-parameter family of dynamic systems with cosymmetry. AB - A study is reported of the bifurcation of the branching of a cycle (Poincare Andronov-Hopf bifurcation) from a smooth one-dimensional submanifold of equilibria of a dynamical system that depends on a vector parameter and admits cosymmetry. The paper reports a topological classification of local phase portraits near a known equilibrium, when the system parameter is close to its critical value that corresponds to an oscillatory instability. New phenomena that are not observed in the classical case of an isolated equilibrium include a delay of cycle creation with respect to the system parameter, loss of stability by the family of equilibria without loss of attraction, and the possibility of unstable supercritical self-oscillations. (c) 1997 American Institute of Physics. PMID- 12779666 TI - The evolution of the magnetic moment in a corrugated magnetic field. AB - In the first part, the equations of motion in a weakly corrugated, periodic magnetic field are linearized and solved by using paraxial approximation, to describe the model and the associated resonance condition. In the second part, the nonlinear evolution of the magnetic moment of resonant particles, in connection with their axial displacement is investigated analytically by using the multiple scale method. It is seen that the linear evolution is converted into a slow and periodic oscillation around the unperturbed value, with a considerable amplitude. The analytic expressions for the period and amplitude of the oscillations are derived and compared with the numerical simulations, which are also presented. Finally, the limitations of the paraxial approximation are concluded by investigating the numerical simulations, with actual field expressions. (c) 1997 American Institute of Physics. PMID- 12779667 TI - Master-slave synchronization and the Lorenz equations. AB - Since the seminal remark by Pecora and Carroll [Phys. Rev. Lett. 64, 821 (1990)] that one can synchronize chaotic systems, the main example in the related literature has been the Lorenz equations. Yet this literature contains a mixture of true and false, and of justified and unsubstantiated claims about the synchronization properties of the Lorenz equations. In this note we clarify some of the confusion. (c) 1997 American Institute of Physics. PMID- 12779668 TI - Synchronization of chaotic systems: Transverse stability of trajectories in invariant manifolds. AB - We examine synchronization of identical chaotic systems coupled in a drive/response manner. A rigorous criterion is presented which, if satisfied, guarantees that synchronization to the driving trajectory is linearly stable to perturbations. An easy to use approximate criterion for estimating linear stability is also presented. One major advantage of these criteria is that, for simple systems, many of the calculations needed to implement them can be performed analytically. Geometrical interpretations of the criterion are discussed, as well as how they may be used to investigate synchronization between mutual coupled systems and the stability of invariant manifolds within a dynamical system. Finally, the relationship between our criterion and results from control theory are discussed. Analytical and numerical results from tests of these criteria on four different dynamical systems are presented. (c) 1997 American Institute of Physics. PMID- 12779669 TI - Detecting noise in a time series. AB - A numerical algorithm is presented for estimating whether, and roughly to what extent, a time series is noise corrupted. Using phase-randomized surrogates constructed from the original signal, metrics are defined which can be used to quantify the noise level. A saturation occurs in these metrics at signal to noise ratios (SNRs) of around 0 dB and below, and also at around 20 dB and above. In between these two regions there is a monotonic transition in the value of the metrics from one region to the other corresponding to changes in the SNR. (c) 1997 American Institute of Physics. PMID- 12779670 TI - Scalar observations from a class of high-dimensional chaotic systems: Limitations of the time delay embedding. AB - The time delay embedding for the reconstruction of a state space from scalar data introduces strong folding of the smooth manifold in which a chaotic attractor is embedded, which is absent in some more natural state space. In order to observe the deterministic nature of data, the typical length scale related to this folding has to be resolved. Above this length scale the data appear to be random. For a particular model class we prove these statements and we derive analytically the dependence of this length scale on the complexity of the system. We show that the number of scalar observations required to observe determinism increases exponentially in the product of the system's entropy and dimension. (c) 1997 American Institute of Physics. PMID- 12779671 TI - Nonlinear analysis of the pharmacological conversion of sustained atrial fibrillation in conscious goats by the class Ic drug cibenzoline. AB - Methods from nonlinear dynamics were applied to test the hypothesis that the dynamics of sustained atrial fibrillation (AF) is modified by the class Ic drug cibenzoline during pharmacological conversion. The experiments were performed in conscious goats in which sustained AF was induced by continuous maintenance of AF via programmed electrical stimulation. Data were collected from electrophysiological experiments in five goats to terminate sustained AF by continuous infusion of cibenzoline. Sets of five unipolar epicardial electrograms of one minute duration were recorded from the left and right atrial free wall during sustained AF (control), and at three episodes during infusion of cibenzoline, when the mean AF interval had been prolonged to 25%, 50% and 85% with respect to control. Ventricular far-field potentials were removed from atrial electrograms by a coherent averaging procedure. Using the Grassberger Procaccia method, the dynamics of the local atrial electrograms was investigated by estimating the (coarse-grained) correlation dimension and correlation entropy from the correlation integral. The results were related to a recently proposed classification (types I-III) of AF based on the degree of complexity of atrial activation patterns. The coarse-grained correlation dimension D(cg) and entropy K(cg) indicated that sustained AF corresponded to type II. During drug administration the coarse-grained parameters were not significantly different from control. Scaling regions in the correlation integral were observed after infusion of cibenzoline (3 out of 5 goats) suggesting that the drug introduced low-dimensional features (type I) in the dynamics of AF (correlation dimension D ranging from 2.8 to 4.4 and correlation entropy K from 1.6 to 6.2 nats/s). Sinus rhythm recorded shortly after cardioversion was very regular (D<2 and K<3 nats/s). The hypothesis that the electrograms during AF and sinus rhythm were generated by a static transformation of a linear Gaussian random process was rejected using a test for time reversibility. The nonlinear analysis revealed that cibenzoline does not significantly alter the dynamics of sustained AF during pharmacological conversion other than a slowing down of the atrial activation and a somewhat increasing global organization of the atrial activation pattern. The sudden change in the dynamical behavior at cardioversion suggests a mechanism that is reminiscent of a bifurcation. (c) 1997 American Institute of Physics. PMID- 12779672 TI - Internal symmetries of cellular automata. AB - (Internal) transformations on the space Sigma of automaton configurations are defined as bi-infinite sequences of permutations of the cell symbols. A pair of transformations (gamma,theta) is said to be an internal symmetry of a cellular automaton f:Sigma-->Sigma if f=theta(-1)fgamma. It is shown that the full group of internal symmetries of an automaton f can be encoded as a group homomorphism F such that theta=F(gamma). The domain and image of the homomorphism F have, in general, infinite order and F is presented by a local automaton-like rule. Algorithms to compute the symmetry homomorphism F and to classify automata by their symmetries are presented. Examples on the types of dynamical implications of internal symmetries are discussed in detail. (c) 1997 American Institute of Physics. PMID- 12779673 TI - Spatiotemporal dynamics in the coherence collapsed regime of semiconductor lasers with optical feedback. AB - This paper presents a spatiotemporal characterization of the dynamics of a single mode semiconductor laser with optical feedback. I use the two-dimensional representation of a time-delayed system (where the delay time plays the role of a space variable) to represent the time evolution of the output intensity and the phase delay in the external cavity. For low feedback levels the laser output is generally periodic or quasiperiodic and with the 2D representation I obtain quasiperiodic patterns. For higher feedback levels the coherence collapsed regime arises, and in the 2D patterns the quasiperiodic structures break and "defects" appear. In this regime the patterns present features that resemble those of an extended spatiotemporally chaotic system. The 2D representation allows the recognition of two distinct types of transition to coherence collapse. As the feedback intensity grows the number of defects increases and the patterns become increasingly chaotic. As the delay time increases the number of defects in the patterns do not increase and there is a signature of the previous quasiperiodic structure that remains. The nature of the two transitions is understood by examining the behavior of various chaotic indicators (the field autocorrelation function, the Lyapunov spectrum, the fractal dimension, and the metric entropy) when the feedback intensity and the delay time vary. (c) 1997 American Institute of Physics. PMID- 12779674 TI - Cellular pattern formation in circular domains. AB - An analysis of stationary and nonstationary cellular patterns observed in premixed flames on a circular, porous plug burner is presented. A phenomenological model is introduced, that exhibits patterns similar to the experimental states. The primary modes of the model are combinations of Fourier Bessel functions, whose radial parts have neighboring zeros. This observation explains several features of patterns, such as the existence of concentric rings of cells and the weak coupling between rings. Properties of rotating rings of cells, including the existence of modulated rotations and heteroclinic cycles can be deduced using mode coupling. For nonstationary patterns, the modal decomposition of experimental data can be carried out using the Karhunen-Loeve (KL) analysis. Experimental states are used to demonstrate the possibility of using KL analysis to differentiate between uniform and nonuniform rotations. The methodology can be extended to study more complicated nonstationary patterns. In particular, it is shown how the complexity of "hopping states" can be unraveled through the analysis. (c) 1997 American Institute of Physics. PMID- 12779675 TI - Bifurcation analysis of the travelling waveform of FitzHugh-Nagumo nerve conduction model equation. AB - The FitzHugh-Nagumo model for travelling wave type neuron excitation is studied in detail. Carrying out a linear stability analysis near the equilibrium point, we bring out various interesting bifurcations which the system admits when a specific Z(2) symmetry is present and when it is not. Based on a center manifold reduction and normal form analysis, the Hopf normal form is deduced. The condition for the onset of limit cycle oscillations is found to agree well with the numerical results. We further demonstrate numerically that the system admits a period doubling route to chaos both in the presence as well as in the absence of constant external stimuli. (c) 1997 American Institute of Physics. PMID- 12779676 TI - Time scales of overdamped nonlinear Brownian motion in arbitrary potential profiles. AB - This paper presents a new approach and new results concerning the problem of the rate of the nonlinear Brownian motion in a field of force. Within the scope of one-dimensional overdamped Brownian motion the exact values of time scales are obtained in detail for three types of arbitrary dimensionless potential profiles varphi(x) which differ from one another by their limiting values at x=+/- infinity. Three time scales, the relaxation time, the decay time of a metastable state and the life time of a nonstable state, are found via given potential profiles varphi(x) by explicit quadrature formulae, as it takes place for the well-known mean first passage times. Specific examples are considered. (c) 1997 American Institute of Physics. PMID- 12779677 TI - Introduction: Control and synchronization of chaos. PMID- 12779678 TI - Targeting in Hamiltonian systems that have mixed regular/chaotic phase spaces. AB - The problem of directing a trajectory of a chaotic dynamical system to a target has been previously considered, and it has been shown that chaos allows targeting using only small controls. In this paper we consider targeting in a Hamiltonian system, whose phase space contains a mixture of regular quasi-periodic and chaotic regions. A multistep forward-backward method targeting strategic intermediate points is found to be efficient and robust. It takes full advantage of the phase space structure and is believed to yield optimal transport times. It is robust under the influence of small noise and small modeling errors and recovers from temporary loss of control. Two illustrative examples, the standard map and the restricted circular three body problem, are presented. (The latter corresponds to motion of a space probe in the presence of the earth and the moon.) Comparisons are made of our method to other targeting strategies. (c) 1997 American Institute of Physics. PMID- 12779679 TI - Fundamentals of synchronization in chaotic systems, concepts, and applications. AB - The field of chaotic synchronization has grown considerably since its advent in 1990. Several subdisciplines and "cottage industries" have emerged that have taken on bona fide lives of their own. Our purpose in this paper is to collect results from these various areas in a review article format with a tutorial emphasis. Fundamentals of chaotic synchronization are reviewed first with emphases on the geometry of synchronization and stability criteria. Several widely used coupling configurations are examined and, when available, experimental demonstrations of their success (generally with chaotic circuit systems) are described. Particular focus is given to the recent notion of synchronous substitution-a method to synchronize chaotic systems using a larger class of scalar chaotic coupling signals than previously thought possible. Connections between this technique and well-known control theory results are also outlined. Extensions of the technique are presented that allow so-called hyperchaotic systems (systems with more than one positive Lyapunov exponent) to be synchronized. Several proposals for "secure" communication schemes have been advanced; major ones are reviewed and their strengths and weaknesses are touched upon. Arrays of coupled chaotic systems have received a great deal of attention lately and have spawned a host of interesting and, in some cases, counterintuitive phenomena including bursting above synchronization thresholds, destabilizing transitions as coupling increases (short-wavelength bifurcations), and riddled basins. In addition, a general mathematical framework for analyzing the stability of arrays with arbitrary coupling configurations is outlined. Finally, the topic of generalized synchronization is discussed, along with data analysis techniques that can be used to decide whether two systems satisfy the mathematical requirements of generalized synchronization. (c) 1997 American Institute of Physics. PMID- 12779680 TI - Control of chaos in excitable physiological systems: A geometric analysis. AB - Model-independent chaos control techniques are inherently well-suited for the control of physiological systems for which quantitative system models are unavailable. The proportional perturbation feedback (PPF) control paradigm, which uses electrical stimulation to perturb directly the controlled system variable (e.g., the interbeat or interspike interval), was developed for excitable physiological systems that do not have an easily accessible system parameter. We develop the stable manifold placement (SMP) technique, a PPF-type technique which is simpler and more robust than the original PPF control algorithm. We use the SMP technique to control a simple geometric model of a chaotic system in the neighborhood of an unstable periodic orbit (UPO). We show that while the SMP technique can control a chaotic system that has UPO dynamics which are characterized by one stable manifold and one unstable manifold, the success of the SMP technique is sensitive to UPO parameter estimation errors. (c) 1997 American Institute of Physics. PMID- 12779681 TI - The OPCL control method for entrainment, model-resonance, and migration actions on multiple-attractor systems. AB - A survey is given of three different control objectives that can be achieved with the use of the Open-Plus-Closed-Loop (OPCL) control method, developed by Jackson and Grosu. For a system that can be characterized by N first-order ordinary differential equations, these objectives are: (1) the asymptotic entrainment of the system's dynamics to a prescribed "goal" dynamics, g(t); (2) an experimental search method to determine an approximate dynamic model; (3) the transferal of the system from one attractor to any "target" attractor. For one class of systems, this may be accomplished without a model, by using only a short-duration record of the natural dynamics in the target attractor, as demonstrated experimentally using the Chua system. (c) 1997 American Institute of Physics. PMID- 12779682 TI - Controlling chaos in a fast diode resonator using extended time-delay autosynchronization: Experimental observations and theoretical analysis. AB - We stabilize unstable periodic orbits of a fast diode resonator driven at 10.1 MHz (corresponding to a drive period under 100 ns) using extended time-delay autosynchronization. Stabilization is achieved by feedback of an error signal that is proportional to the difference between the value of a state variable and an infinite series of values of the state variable delayed in time by integral multiples of the period of the orbit. The technique is easy to implement electronically and it has an all-optical counterpart that may be useful for stabilizing the dynamics of fast chaotic lasers. We show that increasing the weights given to temporally distant states enlarges the domain of control and reduces the sensitivity of the domain of control on the propagation delays in the feedback loop. We determine the average time to obtain control as a function of the feedback gain and identify the mechanisms that destabilize the system at the boundaries of the domain of control. A theoretical stability analysis of a model of the diode resonator in the presence of time-delay feedback is in good agreement with the experimental results for the size and shape of the domain of control. (c) 1997 American Institute of Physics. PMID- 12779683 TI - Sharp diffraction peaks from chaotic structures. AB - Recently various models for spatially chaotic structures have been proposed. We study the diffraction patterns produced by plane chaotic waves incident on one dimensional chaotic point scatterers. The spacing between the scatterers and the dynamics of the incident wave are given by a logistic map or standard map. We find a sharp diffraction peak when the incident dynamics is produced by the same map as the structure of the spatial configuration. The diffraction pattern is symmetric about the incident direction only if the map dynamics is invertible. Diffraction patterns with chaotic incident waves have a large signal-to-noise ratio and are well suited for pattern identification. We discuss possible applications to the scattering of microwaves from aperiodic structures. (c) 1997 American Institute of Physics. PMID- 12779684 TI - Stability analysis of fixed points via chaos control. AB - This paper reviews recent advances in the application of chaos control techniques to the stability analysis of two-dimensional dynamical systems. We demonstrate how the system's response to one or multiple feedback controllers can be utilized to calculate the characteristic multipliers associated with an unstable periodic orbit. The experimental results, obtained for a single and two coupled diode resonators, agree well with the presented theory. (c) 1997 American Institute of Physics. PMID- 12779685 TI - Multistability and the control of complexity. AB - We show how multistability arises in nonlinear dynamics and discuss the properties of such a behavior. In particular, we show that most attractors are periodic in multistable systems, meaning that chaotic attractors are rare in such systems. After arguing that multistable systems have the general traits expected from a complex system, we pass to control them. Our controlling complexity ideas allow for both the stabilization and destabilization of any one of the coexisting states. The control of complexity differs from the standard control of chaos approach, an approach that makes use of the unstable periodic orbits embedded in an extended chaotic attractor. (c) 1997 American Institute of Physics. PMID- 12779686 TI - Maintenance of chaos in a computational model of a thermal pulse combustor. AB - The dynamics of a thermal pulse combustor model are examined. It is found that, as a parameter related to the fuel flow rate is varied, the combustor will undergo a transition from periodic pulsing to chaotic pulsing to a chaotic transient leading to flameout. Results from the numerical model are compared to those obtained from a laboratory-scale thermal pulse combustor. Finally the technique of maintenance (or anticontrol) of chaos is successfully applied to the model, with the result that the operation of the combustor can be continued well into the flameout regime. (c) 1997 American Institute of Physics. PMID- 12779687 TI - Nonlinear prediction, filtering, and control of chemical systems from time series. AB - Prediction, filtering and control of nonlinear systems is formulated in terms of corresponding nonlinear surfaces in the phase space of delayed system readings and control parameters. The construction of these surfaces from time series and their use is demonstrated with a simple chemical model in the chaotic regime. (c) 1997 American Institute of Physics. PMID- 12779688 TI - Adaptive strategies for recognition, noise filtering, control, synchronization and targeting of chaos. AB - Combining knowledge of the local variation rates with some information on the long time trends of a dynamical system, we introduce an adaptive recognition technique consisting in a sequence of variable resolution observation intervals at which the geometrical positions are sampled. The sampling times are chosen so that the sequence of observed points forms a regularized set, in the sense that the separation of adjacent points is almost uniform. We show how this adaptive technique is able to recognize the unstable periodic orbits embedded within a chaotic attractor and stabilize anyone of them even in the presence of noise, through small additive corrections to the dynamics. These techniques have been applied to the synchronization of three chaotic systems, assuring secure communication between a message sender and a message receiver; furthermore they provide robust solutions to the problems of targeting of chaos and of filtering the noise out of an experimental chaotic data set. Implementation of adaptive methods to chaotic Lorenz, three and four dimensional Roessler models and Mackey Glass delayed system are reported.(c) 1997 American Institute of Physics. PMID- 12779689 TI - Synchronizing spatiotemporal chaos. AB - We show analytically and numerically that a pair of uni-directionally coupled spatially extended systems can synchronize. For the case of partial differential equations the synchronization can be achieved by applying the scalar driving signals only at finite number of space points. Our approach is very general and can be useful for practical applications since the synchronization is achieved via feeding in the response system only the information from certain (discrete) spatial locations of the drive system. We also stress some open problems in the field of synchronization of spatiotemporal chaos. (c) 1997 American Institute of Physics. PMID- 12779690 TI - Control and synchronization of chaos in high dimensional systems: Review of some recent results. AB - Controlling chaos and synchronization of chaos have evolved for a number of years as essentially two separate areas of research. Only recently it has been realized that both subjects share a common root in control theory. In addition, as limitations of low dimensional chaotic systems in modeling real world phenomena become increasingly apparent, investigations into the control and synchronization of high dimensional chaotic systems are beginning to attract more interest. We review some recent advances in control and synchronization of chaos in high dimensional systems. Efforts will be made to stress the common origins of the two subjects. (c) 1997 American Institute of Physics. PMID- 12779691 TI - On a simple recursive control algorithm automated and applied to an electrochemical experiment. AB - We review a simple recursive proportional feedback (RPF) control strategy for stabilizing unstable periodic orbits found in chaotic attractors. The method is generally applicable to high-dimensional systems and stabilizes periodic orbits even if they are completely unstable, i.e., have no stable manifolds. The goal of the control scheme is the fixed point itself rather than a stable manifold and the controlled system reaches the fixed point in d+1 steps, where d is the dimension of the state space of the Poincare map. We provide a geometrical interpretation of the control method based on an extended phase space. Controllability conditions or special symmetries that limit the possibility of using a single control parameter to control multiply unstable periodic orbits are discussed. An automated adaptive learning algorithm is described for the application of the control method to an experimental system with no previous knowledge about its dynamics. The automated control system is used to stabilize a period-one orbit in an experimental system involving electrodissolution of copper. (c) 1997 American Institute of Physics. PMID- 12779692 TI - Tracking controlled chaos: Theoretical foundations and applications. AB - Tracking controlled states over a large range of accessible parameters is a process which allows for the experimental continuation of unstable states in both chaotic and non-chaotic parameter regions of interest. In algorithmic form, tracking allows experimentalists to examine many of the unstable states responsible for much of the observed nonlinear dynamic phenomena. Here we present a theoretical foundation for tracking controlled states from both dynamical systems as well as control theoretic viewpoints. The theory is constructive and shows explicitly how to track a curve of unstable states as a parameter is changed. Applications of the theory to various forms of control currently used in dynamical system experiments are discussed. Examples from both numerical and physical experiments are given to illustrate the wide range of tracking applications. (c) 1997 American Institute of Physics. PMID- 12779693 TI - Phase synchronization of chaotic oscillations in terms of periodic orbits. AB - We consider phase synchronization of chaotic continuous-time oscillator by periodic external force. Phase-locking regions are defined for unstable periodic cycles embedded in chaos, and synchronization is described in terms of these regions. A special flow construction is used to derive a simple discrete-time model of the phenomenon. It allows to describe quantitatively the intermittency at the transition to phase synchronization. (c) 1997 American Institute of Physics. PMID- 12779695 TI - Generalized entropies of chaotic maps and flows: A unified approach. AB - A thermodynamic study of nonlinear dynamical systems, based on the orbits' return times to the elements of a generating partition, is proposed. Its grand canonical nature makes it suitable for application to both maps and flows, including autonomous ones. When specialized to the evaluation of the generalized entropies K(q), this technique reproduces a well-known formula for the metric entropy K(1) and clarifies the relationship between a flow and the associated Poincare maps, beyond the straightforward case of periodically forced nonautonomous systems. Numerical estimates of the topological and metric entropy are presented for the Lorenz and Rossler systems. The analysis has been carried out exclusively by embedding scalar time series, ignoring any further knowledge about the systems, in order to illustrate its usefulness for experimental signals as well. Approximations to the generating partitions have been constructed by locating the unstable periodic orbits of the systems up to order 9. The results agree with independent estimates obtained from suitable averages of the local expansion rates along the unstable manifolds. (c) 1997 American Institute of Physics. PMID- 12779694 TI - Sensitive dependence on initial conditions for cellular automata. AB - The property of sensitive dependence on intial conditions is the basis of a rigorous mathematical construction of local maximum Lyapunov exponents for cellular automata. The maximum Lyapunov exponent is given by the fastest average velocity of either the left or right propagating damage fronts. Deviations from the long term behavior of the finite time Lyapunov exponents due to generation of information are quantified and could be used for the characterization of the space time complexity of cellular automata. (c) 1997 American Institute of Physics. PMID- 12779696 TI - Entropy potential and Lyapunov exponents. AB - According to a previous conjecture, spatial and temporal Lyapunov exponents of chaotic extended systems can be obtained from derivatives of a suitable function, the entropy potential. The validity and the consequences of this hypothesis are explored in detail. The numerical investigation of a continuous-time model provides a further confirmation to the existence of the entropy potential. Furthermore, it is shown that the knowledge of the entropy potential allows determining also Lyapunov spectra in general reference frames where the time-like and space-like axes point along generic directions in the space-time plane. Finally, the existence of an entropy potential implies that the integrated density of positive exponents (Kolmogorov-Sinai entropy) is independent of the chosen reference frame. (c) 1997 American Institute of Physics. PMID- 12779697 TI - On chaotic dynamics in "pseudobilliard" Hamiltonian systems with two degrees of freedom. AB - A new class of Hamiltonian dynamical systems with two degrees of freedom is studied, for which the Hamiltonian function is a linear form with respect to moduli of both momenta. For different potentials such systems can be either completely integrable or behave just as normal nonintegrable Hamiltonian systems with two degrees of freedom: one observes many of the phenomena characteristic of the latter ones, such as a breakdown of invariant tori as soon as the integrability is violated; a formation of stochastic layers around destroyed separatrices; bifurcations of periodic orbits, etc. At the same time, the equations of motion are simply integrated on subsequent adjacent time intervals, as in billiard systems; i.e., all the trajectories can be calculated explicitly: Given an initial data, the state of the system is uniquely determined for any moment. This feature of systems in interest makes them very attractive models for a study of nonlinear phenomena in finite-dimensional Hamiltonian systems. A simple representative model of this class (a model with quadratic potential), whose dynamics is typical, is studied in detail. (c) 1997 American Institute of Physics. PMID- 12779698 TI - Waves of excitation on nonuniform membrane rings, caustics, and reverse involutes. AB - Chemical wave experiments on concentric nonuniform membrane rings are presented together with their theoretical description. A new technique is applied to create a slow inner and a fast outer zone in an annular membrane. An abrupt qualitative change of the wave profile was observed while decreasing the wave velocity in the inner zone. This phenomenon and all the experimental wave profiles can be adequately described by assuming that waves are involutes of a relevant caustic. A possible connection with recent models of atrial flutter is also set forth. (c) 1997 American Institute of Physics. PMID- 12779699 TI - Synchronizing Moore and Spiegel. AB - This paper presents a study of bifurcations and synchronization {in the sense of Pecora and Carroll [Phys. Rev. Lett. 64, 821-824 (1990)]} in the Moore-Spiegel oscillator equations. Complicated patterns of period-doubling, saddle-node, and homoclinic bifurcations are found and analyzed. Synchronization is demonstrated by numerical experiment, periodic orbit expansion, and by using coordinate transformations. Synchronization via the resetting of a coordinate after a fixed interval is also successful in some cases. The Moore-Spiegel system is one of a general class of dynamical systems and synchronization is considered in this more general context. (c) 1997 American Institute of Physics. PMID- 12779700 TI - Fractional kinetic equations: solutions and applications. AB - Fractional generalization of the diffusion equation includes fractional derivatives with respect to time and coordinate. It had been introduced to describe anomalous kinetics of simple dynamical systems with chaotic motion. We consider a symmetrized fractional diffusion equation with a source and find different asymptotic solutions applying a method which is similar to the method of separation of variables. The method has a clear physical interpretation presenting the solution in a form of decomposition of the process of fractal Brownian motion and Levy-type process. Fractional generalization of the Kolmogorov-Feller equation is introduced and its solutions are analyzed. (c) 1997 American Institute of Physics. PMID- 12779701 TI - Ring wave solutions of a n+1-dimensional Sine-Gordon model. AB - The dynamical properties of the ring wave solutions of the model psi(tt)- nabla (n) (2)psi+sin psi+ varepsilon sin (psi/2)+alphapsi(t)=0 (00, alpha=0 (or alpha>0) the return effect of the ring wave does not occur only for well defined values of varepsilon. It will be shown numerically that the dissipative perturbation alphapsi(t) (alpha>0) stabilizes both the velocity and the wave profile of the ring wave when the return effect does not occur. (c) 1997 American Institute of Physics. PMID- 12779702 TI - On invariant manifolds attached to oscillating boundaries in Stokes flows. AB - Consider a steady Stokes flow bounded by a flat wall to which a separatrix, not tangent to the wall, is attached at point S. If the wall is made to slowly oscillate periodically with small amplitude in a longitudinal direction, then the Poincare map associated with this perturbed flow has a fixed point on the wall near S with an invariant manifold. This result also holds for flows bounded by a cylindrical wall and flows in which the small periodic perturbation is not due to the oscillation of the wall. This result is applied to several flow geometries in the literature. (c) 1997 American Institute of Physics. PMID- 12779703 TI - Chaotic dynamics of sea clutter. AB - The notion that a deterministic nonlinear dynamical system (with relatively few degrees of freedom) can display aperiodic behavior has a strong bearing on sea clutter characterization: random-looking sea clutter may be the outcome of a chaotic process. This new approach envisages deterministic rules for the underlying sea clutter dynamics, in contrast to the stochastic approach where sea clutter is viewed as a random process with a large number of degrees of freedom. In this paper, we demonstrate, convincingly for the first time, the chaotic dynamics of sea clutter. We say so on the basis of results obtained using radar data collected from a series of extensive and thorough experiments, which have been carried out with ground-truthed sea clutter data sets at three different sites. The study includes correlation dimension analysis (based on the maximum likelihood principle) and Lyapunov spectrum analysis. The Lyapunov (Kaplan-Yorke) dimension, which is a byproduct of Lyapunov spectrum analysis, shows that it is indeed a good estimator of the correlation dimension. The Lyapunov spectrum also reveals that sea clutter is produced by a coupled system of nonlinear differential equations of order five or six. (c) 1997 American Institute of Physics. PMID- 12779704 TI - Dynamics of transients in yttrium-iron-garnet. AB - Yttrium-iron-garnet (YIG) is an important technological material used in microwave devices. In this paper we use dual microwave (1-4 GHz) drives to study the dynamical bifurcation behavior of magnetostatic and spin-wave modes in YIG spheres and rectangular films. The samples are placed in a dc magnetic field and driven by cw and pulse-modulated microwave excitations at magnetostatic mode frequencies. A second microwave drive applied to the sample excites additional spin-wave modes that can interact with those arising from the original excitation and thereby affect the transmission characteristics at the primary frequency. We find a significant decrease in transmission of the primary when the secondary frequency is tuned to approximately half that of the primary drive. This decrease is observed both in the steady state behavior and in the initial overshoot transient associated with pulse modulation of the primary excitation. Results such as these are often treated by extending linear theory to include higher order interaction terms. Herein we present a simple dynamical model that reproduces results that qualitatively resemble the experimental data. (c) 1997 American Institute of Physics. PMID- 12779705 TI - An experimental spatio-temporal state transition of coupled magneto-elastic system. AB - In this paper the vibration and the traveling wave in a coupled magneto-elastic beam system are discussed experimentally. The vibration excited by the periodical forcing at the beam system propagates to another as a wave through the coupling elastic beams. Each magneto-elastic beam shows the variety of vibrations caused by the nonlinearity of the potential well and the wave propagation with time delay. The temporal vibration of the magneto-elastic beam is explained with relations to the spatial state transition based on the experimental results. (c) 1997 American Institute of Physics. PMID- 12779706 TI - Nonannihilation dynamics in an exothermic reaction-diffusion system with mono stable excitability. AB - We consider a 2-component excitable and diffusive system which describes a simple exothermic reaction process. In some parameter regime, there are two characteristics of travelling pulses of the system: (i) travelling pulses are planarly unstable; (ii) when two travelling pulses approach closely, they do not annihilate each other and repel like elastic objects. Under this situation, it is shown that ring patterns break down into complex patterns in 2-dimensions, which are totally different from those arising in the well-known excitable and diffusive system with the FitzHugh-Nagumo nonlinearity. (c) 1997 American Institute of Physics. PMID- 12779707 TI - Evolving perspectives during 12 years of electrical turbulence. AB - This Focus issue describes a problem in electrical dynamics which has fascinated generations of physiologists. There are today so many views of fibrillation that only the rarest generalization can embrace all of them. Fifty-two prominent investigators collaborate here to present aspects of the problem in these eighteen articles (including this introduction) tailored for readers whose principal expertise lies elsewhere. In "The High One's Lay" (Norse Runes, ca. 800) Odin remarks, "Much too early I came to many places: the beer was not yet ready, or was already drunk em leader " but to this one we come at very nearly the right time in 1998. This introduction attempts to guide newcomers by noting the changed or multiple meanings of novel technical terms while sorting the key facts and ideas into an order that facilitates comparison and contrast with those of a dozen years ago. This Focus issue is authored by some of the foremost innovators of both theory and experiment in this area. By assimilating their presentations the readers of Chaos can become well poised to appreciate and evaluate the definitive evidence expected in the next few years. (c) 1998 American Institute of Physics. PMID- 12779708 TI - Vortex dynamics in three-dimensional continuous myocardium with fiber rotation: Filament instability and fibrillation. AB - Wave propagation in ventricular muscle is rendered highly anisotropic by the intramural rotation of the fiber. This rotational anisotropy is especially important because it can produce a twist of electrical vortices, which measures the rate of rotation (in degree/mm) of activation wavefronts in successive planes perpendicular to a line of phase singularity, or filament. This twist can then significantly alter the dynamics of the filament. This paper explores this dynamics via numerical simulation. After a review of the literature, we present modeling tools that include: (i) a simplified ionic model with three membrane currents that approximates well the restitution properties and spiral wave behavior of more complex ionic models of cardiac action potential (Beeler-Reuter and others), and (ii) a semi-implicit algorithm for the fast solution of monodomain cable equations with rotational anisotropy. We then discuss selected results of a simulation study of vortex dynamics in a parallelepipedal slab of ventricular muscle of varying wall thickness (S) and fiber rotation rate (theta(z)). The main finding is that rotational anisotropy generates a sufficiently large twist to destabilize a single transmural filament and cause a transition to a wave turbulent state characterized by a high density of chaotically moving filaments. This instability is manifested by the propagation of localized disturbances along the filament and has no previously known analog in isotropic excitable media. These disturbances correspond to highly twisted and distorted regions of filament, or "twistons," that create vortex rings when colliding with the natural boundaries of the ventricle. Moreover, when sufficiently twisted, these rings expand and create additional filaments by further colliding with boundaries. This instability mechanism is distinct from the commonly invoked patchy failure or wave breakup that is not observed here during the initial instability. For modified Beeler-Reuter-like kinetics with stable reentry in two dimensions, decay into turbulence occurs in the left ventricle in about one second above a critical wall thickness in the range of 4-6 mm that matches experiment. However this decay is suppressed by uniformly decreasing excitability. Specific experiments to test these results, and a method to characterize the filament density during fibrillation are discussed. Results are contrasted with other mechanisms of fibrillation and future prospects are summarized. (c)1998 American Institute of Physics. PMID- 12779709 TI - Reentrant waves and their elimination in a model of mammalian ventricular tissue. AB - The vulnerability to reentrant wave propagation, its characteristics (period, meander, and stability), the effects of rotational transmural anisotropy, and the control of reentrant waves by small amplitude perturbations and large amplitude defibrillating shocks are investigated theoretically and numerically for models based on high order, stiff biophysically derived excitation equations. PMID- 12779710 TI - Spiral breakup as a model of ventricular fibrillation. AB - The phenomenon of spiral breakup in a 2D and a 3D excitable medium is described. Differences between breakup in two dimensions and in three dimensions are discussed. Spiral breakup in an anatomical model of the ventricles of the heart is also studied. The patterns of excitation in the heart are presented at different wavelengths together with their electrocardiograms. Finally it is suggested that the phenomenon of spiral breakup is a possible mechanism of the ventricular fibrillation (VF). (c) 1998 American Institute of Physics. PMID- 12779711 TI - Ventricular fibrillation and atrial fibrillation are two different beasts. AB - Although the mechanisms of fibrillation are no doubt multi-faceted, the geometry of the heart may play a major role in the dynamics of wave propagation during fibrillation [A. T. Winfree, Science 266, 1003-1006 (1994)]. The ventricles are thick chambers made up of sheets of parallel muscle fibers with the direction of fibers rotating across the ventricular walls (rotational anisotropy). The thick walls of the ventricles allow reentry to develop transmurally, provided the wavelength is sufficiently small. Depending on the kinetics of heart cells, the dynamics of rotating waves in three dimensions may be fundamentally different than in two dimensions, leading to destabilization of reentry and ventricular fibrillation (VF) in thick ventricles. The atria have an intricate geometry comprised of a thin sheet of cardiac tissue attached to a very complex network of pectinate muscles. The branching geometry of the pectinate muscles may lead to destabilization of two-dimensional reentry via "long-distance" electrical connections giving rise to atrial fibrillation (AF). Therefore, although fibrillation occurs via complex three-dimensional wave propagation in the ventricles and the atria, the underlying mechanisms and factors that sustain VF and AF are probably different.(c) 1998 American Institute of Physics. PMID- 12779712 TI - Self-organization and the dynamical nature of ventricular fibrillation. AB - This article reviews recent data supporting the conjecture that, in the structurally and electrophysiologically normal heart, cardiac fibrillation is not a totally random phenomenon. Experimental and numerical studies based on the theory of excitable media suggest that fibrillation in the mammalian ventricles is the result of self-organized three-dimensional (3-D) electrical rotors giving rise to scroll waves that move continuously (i.e., drift) throughout the heart at varying speeds. A brief review of studies on the dynamics of rotors in two dimensional (2-D) and 3-D excitable media is presented with emphasis on the experimental demonstration of such dynamics in cardiac muscle of various species. The discussion is centered on rotor dynamics in the presence and the absence of structural heterogeneities, and in the phenomena of drifting and anchoring, which in the electrocardiogram (ECG) may manifest as life-threatening cardiac rhythm disturbances. For instance, in the rabbit heart, a single electrical rotor that drifts rapidly throughout the ventricles gives rise to complex patterns of excitation. In the ECG such patterns are indistinguishable from ventricular fibrillation. On the other hand, a rotor that anchors to a discontinuity or defect in the muscle (e.g., a scar, a large artery or a bundle of connective tissue) may result in stationary rotating activity, which in the ECG is manifested as a form of so-called "monomorphic" ventricular tachycardia. More recent data show that ventricular fibrillation occurs in mammals irrespective of size or species. While in small hearts, such as those of mice and rabbits, a single drifting or meandering rotor can result in fibrillation, in larger hearts, such as the sheep and possibly the human, fibrillation occurs in the form of a relatively small number of coexisting but short-lived rotors. Overall, the work discussed here has paved the way for a better understanding of the mechanisms of fibrillation in the normal, as well as diseased human heart. (c) 1998 American Institute of Physics. PMID- 12779713 TI - A method for visualization of ventricular fibrillation: Design of a cooled fiberoptically coupled image intensified CCD data acquisition system incorporating wavelet shrinkage based adaptive filtering. AB - The measurement of cardiac transmembrane potential changes with voltage sensitive dyes is in increasing use. Detection of these very small fluorescent alterations using large multiplexed arrays, such as charge coupled device (CCD) cameras at high sampling rates, has proven challenging and usually requires significant averaging to improve the signal-to-noise ratio. To minimize the damage of living tissue stained with voltage sensitive dyes, excitation photon exposure must be limited, with the inevitable consequence of diminishing the fluorescence that is generated. State-of-the-art high frame rate CCD cameras have read noise levels in the 5-10 e(-) rms range, which is at least two orders of magnitude above that required to detect voltage sensitive dye alterations at individual pixels corresponding to 1 mm(2) heart regions illuminated with levels of 100 mW/cm(2) at frame rates approaching 1000 frames/sec. Image intensification is thus required prior to photon quantification. We report here the development of such a data acquisition system using commercially available hardware. Additionally, in the past ten years, a mathematical theory of multiresolution has been developed, and new building blocks called wavelets, allow a signal to be observed at different resolutions. Wavelet analysis also makes possible a new method of extricating signals from noise. We have incorporated spatially adaptive filters based on wavelet denoising of individual pixels to significantly reduce the multiple noise sources present in the acquired data. (c) 1998 American Institute of Physics. PMID- 12779714 TI - Spatial organization, predictability, and determinism in ventricular fibrillation. AB - The degree of spatial organization of ventricular fibrillation (VF) is a fundamental dynamical property of the arrhythmia and may determine the success of proposed therapeutic approaches. Spatial organization is closely related to the dimension of VF, and hence to its predictability and controllability. We have explored several techniques to quantify spatial organization during VF, to predict patterns of activity, and to see how spatial organization and predictability change as the arrhythmia progresses. Epicardial electrograms recorded from pig hearts using rectangular arrays of unipolar extracellular electrodes (1 mm spacing) were analyzed. The correlation length of VF, the number of Karhunen-Loeve modes required to approximate data during VF, the number, size and recurrence of wavefronts, and the mean square error of epicardial potential fields predicted 0.256 seconds into the future were all estimated. The ability of regularly-timed pacing stimuli to capture areas of fibrillating myocardium during VF was confirmed by a significant increase in local spatial organization. Results indicate that VF is neither "low-dimensional chaos" (dimension <5) nor "random" behavior (dimension= infinity ), but is a high-dimensional response with a degree of spatial coherence that changes as the arrhythmia progresses. (c) 1998 American Institute of Physics. PMID- 12779715 TI - Electrical turbulence as a result of the critical curvature for propagation in cardiac tissue. AB - In cardiac tissue, the propagation of electrical excitation waves is dependent on the active properties of the cell membrane (ionic channels) and the passive electrical properties of cardiac tissue (passive membrane properties, distribution of gap junctions, and cell shapes). Initiation of cardiac arrhythmias is usually associated with heterogeneities in the active and/or passive properties of cardiac tissue. However, as a result of the effect of wave front geometry (curvature) on propagation of cardiac waves, inexcitable anatomical obstacles, like veins and arteries, may cause the formation of self sustained vortices and uncontrolled high-frequency excitation in normal homogeneous myocardium. (c) 1998 American Institute of Physics. PMID- 12779716 TI - Computerized mapping of fibrillation in normal ventricular myocardium. AB - It is well known that the ability to fibrillate is intrinsic to a normal ventricle that exceeds a critical mass. The questions we address are how is ventricular fibrillation (VF) initiated and perpetuated in normal myocardium, and why is VF not seen more often in the general population if all ventricles have the ability to fibrillate. To study the mechanisms of VF, we used computerized mapping techniques with up to 512 channels of simultaneous multisite recordings for data acquisition. The data were then processed for dynamic display of the activation patterns and for mathematical analyses of the activation intervals. The results show that in normal ventricles, VF can be initiated by a single strong premature stimulus given during the vulnerable period of the cardiac cycle. The initial activations form a figure-eight pattern. Afterward, VF will perpetuate itself without any outside help. The self-perpetuation itself is due to at least two factors. One is that single wave fronts spontaneously break up into two or more wavelets. The second is that when two wavelets intersect perpendicular to each other, the second wavelet is broken by the residual refractoriness left over from the first wavelet. Mathematical analyses of the patterns of activation during VF revealed that VF is a form of chaos, and that transition from ventricular tachycardia (VT) to VF occurs via the quasiperiodic route. In separate experiments, we found that we can convert VF to VT by tissue size reduction. The physiological mechanism associated with the latter transition appears to be the reduction of the number of reentrant wave fronts and wandering wavelets. Based on these findings, we propose that the reentrant wave fronts and the wandering wavelets serve as the physiological equivalent of coupled oscillators. A minimal number of oscillators is needed for VF to perpetuate itself, and to generate chaotic dynamics; hence a critical mass is required to perpetuate VF. We conclude that VF in normal myocardium is a form of reentrant cardiac arrhythmia. A strong electrical stimulus initiates single or dual reentrant wave fronts that break up into multiple wavelets. Sometimes short-lived reentry is also generated during the course of VF. These organized reentrant and broken wavelets serve as coupled oscillators that perpetuate VF and maintain chaos. Although the ability to support these oscillators exists in a normal ventricle, the triggers required to generate them are nonexistent in the normal heart. Therefore, VF and sudden death do not happen to most people with normal ventricular myocardium. (c) 1998 American Institute of Physics. PMID- 12779717 TI - Patterns of spiral tip motion in cardiac tissues. AB - In support of the spiral wave theory of reentry, simulation studies and animal models have been utilized to show various patterns of spiral wave tip motion such as meandering and drifting. However, the demonstration of these or any other patterns in cardiac tissues have been limited. Whether such patterns of spiral tip motion are commonly observed in fibrillating cardiac tissues is unknown, and whether such patterns form the basis of ventricular tachycardia or fibrillation remain debatable. Using a computerized dynamic activation display, 108 episodes of atrial and ventricular tachycardia and fibrillation in isolated and intact canine cardiac tissues, as well as in vitro swine and myopathic human cardiac tissues, were analyzed for patterns of nonstationary, spiral wave tip motion. Among them, 46 episodes were from normal animal myocardium without pharmacological perturbations, 50 samples were from normal animal myocardium, either treated with drugs or had chemical ablation of the subendocardium, and 12 samples were from diseased human hearts. Among the total episodes, 11 of them had obvious nonstationary spiral tip motion with a life span of >2 cycles and with consecutive reentrant paths distinct from each other. Four patterns were observed: (1) meandering with an inward petal flower in 2; (2) meandering with outward petals in 5; (3) irregularly concentric in 3 (core moving about a common center); and (4) drift in 1 (linear core movement). The life span of a single nonstationary spiral wave lasted no more than 7 complete cycles with a mean of 4.6+/-4.3, and a median of 4.5 cycles in our samples. Conclusion: (1) Patently evident nonstationary spiral waves with long life spans were uncommon in our sample of mostly normal cardiac tissues, thus making a single meandering spiral wave an unlikely major mechanism of fibrillation in normal ventricular myocardium. (2) A tendency toward four patterns of nonstationary spiral tip motion was observed. (c) 1998 American Institute of Physics. PMID- 12779718 TI - Vulnerability to ventricular fibrillation. AB - One of the factors that favors the development of ventricular fibrillation is an increase in the dispersion of refractoriness. Experiments will be described in which an increase in dispersion in the recovery of excitability was determined during brief episodes of enhanced sympathetic nerve activity, known to increase the risk of fibrillation. Whereas in the normal heart ventricular fibrillation can be induced by a strong electrical shock, a premature stimulus of moderate intensity only induces fibrillation in the presence of regional ischemia, which greatly increases the dispersion of refractoriness. One factor that is of importance for the transition of reentrant ventricular tachycardia to ventricular fibrillation during acute regional ischemia is the subendocardial Purkinje system. After selective destruction of the Purkinje network by lugol, reentrant tachycardias still develop in the ischemic region, but they do not degenerate into fibrillation. Finally, attempts were made to determine the minimal mass of thin ventricular myocardium required to sustain fibrillation induced by burst pacing. This was done by freezing of subendocardial and midmural layers. The rim of surviving epicardial muscle had to be larger than 20 g. Extracellular electrograms during fibrillation in both the intact and the "frozen" left ventricle were indistinguishable, but activation patterns were markedly different. In the intact ventricle epicardial activation was compatible with multiple wavelet reentry, in the "frozen" heart a single, or at most two wandering reentrant waves were seen. (c) 1998 American Institute of Physics. PMID- 12779719 TI - Two forms of spiral-wave reentry in an ionic model of ischemic ventricular myocardium. AB - It is well known that there is considerable spatial inhomogeneity in the electrical properties of heart muscle, and that the many interventions that increase this initial degree of inhomogeneity all make it easier to induce certain cardiac arrhythmias. We consider here the specific example of myocardial ischemia, which greatly increases the electrical heterogeneity of ventricular tissue, and often triggers life-threatening cardiac arrhythmias such as ventricular tachycardia and ventricular fibrillation. There is growing evidence that spiral-wave activity underlies these reentrant arrhythmias. We thus investigate whether spiral waves might be induced in a realistic model of inhomogeneous ventricular myocardium. We first modify the Luo and Rudy [Circ. Res. 68, 1501-1526 (1991)] ionic model of cardiac ventricular muscle so as to obtain maintained spiral-wave activity in a two-dimensional homogeneous sheet of ventricular muscle. Regional ischemia is simulated by raising the external potassium concentration ([K(+)](o)) from its nominal value of 5.4 mM in a subsection of the sheet, thus creating a localized inhomogeneity. Spiral-wave activity is induced using a pacing protocol in which the pacing frequency is gradually increased. When [K(+)](o) is sufficiently high in the abnormal area (e.g., 20 mM), there is complete block of propagation of the action potential into that area, resulting in a free end or wave break as the activation wave front encounters the abnormal area. As pacing continues, the free end of the activation wave front traveling in the normal area increasingly separates or detaches from the border between normal and abnormal tissue, eventually resulting in the formation of a maintained spiral wave, whose core lies entirely within an area of normal tissue lying outside of the abnormal area ("type I" spiral wave). At lower [K(+)](o) (e.g., 10.5 mM) in the abnormal area, there is no longer complete block of propagation into the abnormal area; instead, there is partial entrance block into the abnormal area, as well as exit block out of that area. In this case, a different kind of spiral wave (transient "type II" spiral wave) can be evoked, whose induction involves retrograde propagation of the action potential through the abnormal area. The number of turns made by the type II spiral wave depends on several factors, including the level of [K(+)](o) within the abnormal area and its physical size. If the pacing protocol is changed by adding two additional stimuli, a type I spiral wave is instead produced at [K(+)](o)=10.5 mM. When pacing is continued beyond this point, apparently aperiodic multiple spiral-wave activity is seen during pacing. We discuss the relevance of our results for arrythmogenesis in both the ischemic and nonischemic heart. (c) 1998 American Institute of Physics. PMID- 12779720 TI - The effect of gap junctional distribution on defibrillation. AB - We summarize a mathematical theory for direct activation and defibrillation of cardiac tissue. We show that the direct stimulus and defibrillation thresholds are likely to be strongly affected by the gap junctional distribution and density, suggesting an indirect experimental test of the theory. (c) 1998 American Institute of Physics. PMID- 12779721 TI - Models of defibrillation of cardiac tissue. AB - Heterogeneities, such as gap junctions, defects in periodical cellular lattices, intercellular clefts and fiber curvature allow one to understand the effect of an electric field in cardiac tissue. They induce membrane potential variations even in the bulk of the myocardium, with a characteristic sawtooth shape. The sawtooth potential, induced by heterogeneities at large scales (tissue strands) can be more easily observed, and lead to stronger effects than the one induced at the cellular level. In the generic model of propagation in cardiac tissue (FitzHugh), 4 mechanisms of defibrillation were found, two mechanisms based on excitation (E(A),E(M)), and two-on de-excitation (D(A),D(M)). The lowest electric field is required by an E(M) mechanism. In the Beeler-Reuter ionic model, mechanism D(M) is impossible. We critically review the experimental basis of the theory and propose new experiments. (c) 1998 American Institute of Physics. PMID- 12779722 TI - The induction of reentry in cardiac tissue. The missing link: How electric fields alter transmembrane potential. AB - This review examines the initiation of reentry in cardiac muscle by strong electric shocks. Specifically, it concentrates on the mechanisms by which electric shocks change the transmembrane potential of the cardiac membrane and create the physiological substrate required by the critical point theory for the initiation of rotors. The mechanisms examined include (1) direct polarization of the tissue by the stimulating current, as described by the one-dimensional cable model and its two- and three-dimensional extensions, (2) the presence of virtual anodes and cathodes, as described by the bidomain model with unequal anisotropy ratios of the intra- and extracellular spaces, (3) polarization of the tissue due to changing orientation of cardiac fibers, and (4) polarization of individual cells or groups of cells by the electric field ("sawtooth potential"). The importance of these mechanisms in the initiation of reentry is examined in two case studies: the induction of rotors using successive stimulation with a unipolar electrode, and the induction of rotors using cross-field stimulation. These cases reveal that the mechanism by which a unipolar stimulation induces arrhythmias can be explained in the framework of the bidomain model with unequal anisotropy ratios. In contrast, none of the examined mechanisms provide an adequate explanation for the induction of rotors by cross-field stimulation. Hence, this study emphasizes the need for further experimental and theoretical work directed toward explaining the mechanism of field stimulation. (c) 1998 American Institute of Physics. PMID- 12779723 TI - The role of cardiac tissue structure in defibrillation. AB - The purpose of this paper is to investigate the relationship between cardiac tissue structure, applied electric field, and the transmembrane potential induced in the process of defibrillation. It outlines a general understanding of the structural mechanisms that contribute to the outcome of a defibrillation shock. Electric shocks defibrillate by changing the transmembrane potential throughout the myocardium. In this process first and foremost the shock current must access the bulk of myocardial mass. The exogenous current traverses the myocardium along convoluted intracellular and extracellular pathways channeled by the tissue structure. Since individual fibers follow curved pathways in the heart, and the fiber direction rotates across the ventricular wall, the applied current perpetually engages in redistribution between the intra- and extracellular domains. This redistribution results in changes in transmembrane potential (membrane polarization): regions of membrane hyper- and depolarization of extent larger than a single cell are induced in the myocardium by the defibrillation shock. Tissue inhomogeneities also contribute to local membrane polarization in the myocardium which is superimposed over the large-scale polarization associated with the fibrous organization of the myocardium. The paper presents simulation results that illustrate various mechanisms by which cardiac tissue structure assists the changes in transmembrane potential throughout the myocardium. (c) 1998 American Institute of Physics. PMID- 12779724 TI - A numerical method for the solution of the bidomain equations in cardiac tissue. AB - A numerical scheme for efficient integration of the bidomain model of action potential propagation in cardiac tissue is presented. The scheme is a mixed implicit-explicit scheme with no stability time step restrictions and requires that only linear systems of equations be solved at each time step. The method is faster than a fully explicit scheme and there is no increase in algorithmic complexity to use this method instead of a fully explicit method. The speedup factor depends on the timestep size, which can be set solely on the basis of the demands for accuracy. (c) 1998 American Institute of Physics. PMID- 12779725 TI - Complexity lower bounds for polygonal billiards. AB - Polygonal billiards have zero topological entropy. Complexity is a finer measure of their asymptotic behavior. In this article we show an explicit quadratic lower bound for the complexity of the billiard in an arbitrary polygon. (c) 1998 American Institute of Physics. PMID- 12779726 TI - Diffusion in Hamiltonian systems. AB - The study is reported of a diffusion in a model of degenerate Hamiltonian systems. The Hamiltonian under consideration is the sum of a linear function of action variables and a periodic function of angle variables. Under certain choices of these functions the diffusion of action variables exists. In the case of two degrees of freedom during the process of diffusion, the vector of the action variables returns many times near its initial value. In the case of three degrees of freedom the choice of Hamiltonian allows one to obtain a diffusion rate faster than any prescribed one. (c) 1998 American Institute of Physics. PMID- 12779727 TI - Motion in a stochastic layer described by symbolic dynamics. AB - The motion in the stochastic layer surrounding an island can be studied by using the standard map: This problem is of direct relevance to the diffusion of magnetic field lines in a tokamak. In a previous work it was shown that this process can be adequately modelled by a continuous time random walk (CTRW) describing transitions of the running point between three basins representing, respectively, trapped motion around the island, and passing motion above or below the island. The sticking property of the island deeply modifies the nature of the transport process, leading to subdiffusive behavior. In the present work it is shown that the motion can be analyzed in terms of a symbolic dynamics which leads to the possibility of an automatic measurement of the data necessary for the construction of the CTRW. The logical features of the procedure are described, and the method is applied to an analysis of long time series, thus completing the results of the previous work. (c) 1998 American Institute of Physics. PMID- 12779728 TI - Solitary wave solutions of nonlocal sine-Gordon equations. AB - In this paper a nonlocal generalization of the sine-Gordon equation, u(tt)+sin u=( partial differential / partial differential x) integral (- infinity ) (+ infinity )G(x-x('))u(x(') )(x('),t)dx(') is considered. We present a brief review of the applications of such equations and show that involving such a nonlocality can change features of the model. In particular, some solutions of the sine Gordon model (for example, traveling 2pi-kink solutions) may disappear in the nonlocal model; furthermore, some new classes of solutions such as traveling topological solitons with topological charge greater than 1 may arise. We show that the lack of Lorenz invariancy of the equation under consideration can lead to a phenomenon of discretization of kink velocities. We discussed this phenomenon in detail for the special class of kernels G(xi)= summation operator (j=1) (N)kappa(j)e(-eta(j)mid R:ximid R:), eta(j)>0, j=1,2, em leader,N. We show that, generally speaking, in this case the velocities of kinks (i) are determined unambiguously by a type of kink and value(s) of kernel parameter(s); (ii) are isolated i.e., if c(*) is the velocity of a kink then there are no other kink solutions of the same type with velocity c in (c(*)- varepsilon,c(*)+ varepsilon ) for a certain value of varepsilon. We also used this special class of kernels to construct approximations for analytical and numerical study of the problem in a more general case. Finally, we set forth results of the numerical investigation of the problem with the kernel that is the McDonald function G(xi) approximately K(0)(mid R:ximid R:/lambda) (lambda is a parameter) that have applications in the Josephson junction theory. (c) 1998 American Institute of Physics. PMID- 12779729 TI - On the absence of a real-analytic first integral for ABC flow when A=B. AB - In this paper we report on the absence of a real-analytic first integral for the well-known system describing the steady-state flow of an ideal incompressible liquid with periodic boundary conditions, called ABC flow, in the case when two of the three parameters entering the system are equal and some inequality exists between them and the third parameter. (c) 1998 American Institute of Physics. PMID- 12779730 TI - Multirhythmic bursting. AB - A complex modeled bursting neuron [C. C. Canavier, J. W. Clark, and J. H. Byrne, J. Neurophysiol. 66, 2107-2124 (1991)] has been shown to possess seven coexisting limit cycle solutions at a given parameter set [Canavier et al., J. Neurophysiol 69, 2252-2259 (1993); 72, 872-882 (1994)]. These solutions are unique in that the limit cycles are concentric in the space of the slow variables. We examine the origin of these solutions using a minimal 4-variable bursting cell model. Poincare maps are constructed using a saddle-node bifurcation of a fast subsystem such as our Poincare section. This bifurcation defines a threshold between the active and silent phases of the burst cycle in the space of the slow variables. The maps identify parameter spaces with single limit cycles, multiple limit cycles, and two types of chaotic bursting. To investigate the dynamical features which underlie the unique shape of the maps, the maps are further decomposed into two submaps which describe the solution trajectories during the active and silent phases of a single burst. From these findings we postulate several necessary criteria for a bursting model to possess multiple stable concentric limit cycles. These criteria are demonstrated in a generalized 3-variable model. Finally, using a less direct numerical procedure, similar return maps are calculated for the original complex model [C. C. Canavier, J. W. Clark, and J. H. Byrne, J. Neurophysiol. 66, 2107-2124 (1991)], with the resulting mappings appearing qualitatively similar to those of our 4-variable model. These multistable concentric bursting solutions cannot occur in a bursting model with one slow variable. This type of multistability arises when a bursting system has two or more slow variables and is viewed as an essentially second-order system which receives discrete perturbations in a state-dependent manner. (c) 1998 American Institute of Physics. PMID- 12779731 TI - Single-head spin modes in frontal polymerization. AB - The single-head spin mode is the first two-dimensional pattern of self propagating thermal reaction fronts observed after the planar front loses stability. A "hot spot" is observed to propagate around the periphery of the front. The dynamics of the single-head frontal polymerization regime were studied experimentally, and two novel results were found. The "hot spot" was measured and found to be superadiabatic. The relationship between the rotational velocity and the propagating velocity was found. Experimental data were also compared with the theoretical results based on the linear stability analysis and found to be in a reasonable agreement. (c) 1998 American Institute of Physics. PMID- 12779732 TI - Easy-to-implement method to target nonlinear systems. AB - In this work we present a method to rapidly direct a chaotic system, to an aimed state or target, through a sequence of control perturbations, with few different amplitudes chosen according to the allowed control-parameter changes. We applied this procedure to the one-dimensional Logistic map, to the two-dimensional Henon map, and to the Double Scroll circuit described by a three-dimensional system of differential equations. Furthermore, for the Logistic map, we show numerically that the resulting trajectory (from the starting point to the target) goes along a stable manifold of the target. Moreover, using the Henon map, we create and stabilize unstable periodic orbits, and also verify the procedure robustness in the presence of noise. We apply our method to the Double Scroll circuit, without using any low-dimensional mapping to represent its dynamics, an improvement with respect to previous targeting methods only applied for experimental systems that are mapping-modeled. (c) 1998 American Institute of Physics. PMID- 12779734 TI - Chaos and irreversibility: Introductory Comments. PMID- 12779733 TI - Synchronization and control of spatiotemporal chaos using time-series data from local regions. AB - In this paper we show that the analysis of the dynamics in localized regions, i.e., sub-systems can be used to characterize the chaotic dynamics and the synchronization ability of the spatiotemporal systems. Using noisy scalar time series data for driving along with simultaneous self-adaptation of the control parameter representative control goals like suppressing spatiotemporal chaos and synchronization of spatiotemporally chaotic dynamics have been discussed. (c) 1998 American Institute of Physics. PMID- 12779735 TI - Probabilistic and thermodynamic aspects of dynamical systems. AB - The probabilistic approach to dynamical systems giving rise to irreversible behavior at the macroscopic, mesoscopic, and microscopic levels of description is outlined. Signatures of the complexity of the underlying dynamics on the spectral properties of the Liouville, Frobenius-Perron, and Fokker-Planck operators are identified. Entropy and entropy production-like quantities are introduced and the connection between their properties in nonequilibrium steady states and the characteristics of the dynamics in phase space are explored. (c) 1998 American Institute of Physics. PMID- 12779736 TI - Thermostats: Analysis and application. AB - Gaussian isokinetic and isoenergetic deterministic thermostats are reviewed in the correct historical context with their later justification using Gauss' principle of least constraint. The Nose-Hoover thermostat for simulating the canonical ensemble is also developed. For some model systems the Lyapunov exponents satisfy the conjugate pairing rule and a Hamiltonian formulation is obtained. We prove the conjugate pairing rule for nonequilibrium systems where the force is derivable from a potential. The generalized symplectic structure and Hamiltonian formulation is discussed. The application of such thermostats to the Lorentz gas is considered in some detail. The periodic orbit expansion methods are used to calculate averages and to categorize the generic transitions in the structure of the attractor. We prove that the conductivity in the nonequilibrium Lorentz gas is non-negative. (c) 1998 American Institute of Physics. PMID- 12779737 TI - The conjugate-pairing rule for non-Hamiltonian systems. AB - In systems that satisfy the Conjugate Pairing Rule (CPR), the spectrum of Lyapunov exponents is symmetric. The sum of each conjugate pair of exponents is identical. Since in dissipative systems the sum of all the exponents is the entropy production divided by Boltzmann's constant, the calculation of transport coefficients from the Lyapunov exponents is greatly simplified in systems that satisfy CPR. Sufficient conditions for CPR are well known: the underlying adiabatic dynamics should be symplectic. However, the necessary conditions for CPR are not known. In this paper we report on the results of computer simulations which shed light on the necessary conditions for the CPR to hold. We provide, for the first time, convincing evidence that the standard molecular dynamics algorithm for calculating shear viscosity violates the CPR, even in the thermodynamic limit. In spite of this it appears that the sum of the maximal exponents is equal to the entropy production per degree of freedom. Thus it appears that the shear viscosity can still be calculated using the standard viscosity algorithm by summing the maximal pair of exponents.(c) 1998 American Institute of Physics. PMID- 12779738 TI - Variational principles for thermostatted systems. AB - A generalized Lagrangian formalism is proposed for dealing with interacting many body systems subject to time reversible friction forces. The corresponding generalized Hamiltonian and Poissonian formalisms are presented. Explicit connection is made between the new Poisson brackets obtained and the 2-forms of conformally symplectic systems recently investigated. Applications to Nose-Hoover and isokinetic dynamics are treated in detail. (c) 1998 American Institute of Physics. PMID- 12779739 TI - Note on phase space contraction and entropy production in thermostatted Hamiltonian systems. AB - The phase space contraction and the entropy production rates of Hamiltonian systems in an external field, thermostatted to obtain a stationary state, are considered. While for stationary states with a constant kinetic energy the two rates are formally equal for all numbers of particles N, for stationary states with constant total (kinetic and potential) energy this only obtains for large N. However, in both cases a large number of particles is required to obtain equality with the entropy production rate of Irreversible Thermodynamics. Consequences of this for the positivity of the transport coefficients and for the Onsager relations are discussed. Numerical results are presented for the special case of the Lorentz gas. (c) 1998 American Institute of Physics. PMID- 12779740 TI - Chaos and irreversibility in simple model systems. AB - The multifractal link between chaotic time-reversible mechanics and thermodynamic irreversibility is illustrated for three simple chaotic model systems: the Baker Map, the Galton Board, and many-body color conductivity. By scaling time, or the momenta, or the driving forces, it can be shown that the dissipative nature of the three thermostated model systems has analogs in conservative Hamiltonian and Lagrangian mechanics. Links between the microscopic nonequilibrium Lyapunov spectra and macroscopic thermodynamic dissipation are also pointed out. (c) 1998 American Institute of Physics. PMID- 12779741 TI - Dynamical ensembles in nonequilibrium statistical mechanics and their representations. AB - The stationary states of driven systems of particles are considered from the point of view of the invariant probability distributions in the phase space which characterize them. The main features of various representations of such distributions are reviewed, and a brief derivation of the one based on orbital measures is given. We mention the limits of the mathematical derivations, and discuss the expected range of applicability beyond such limits. (c) 1998 American Institute of Physics. PMID- 12779742 TI - Chaotic dynamics, fluctuations, nonequilibrium ensembles. AB - The ideas and the conceptual steps leading from the ergodic hypothesis for equilibrium statistical mechanics to the chaotic hypothesis for equilibrium and nonequilibrium statistical mechanics are illustrated. The fluctuation theorem linear law and universal slope prediction for reversible systems is briefly derived. Applications to fluids are briefly alluded to. (c) 1998 American Institute of Physics. PMID- 12779743 TI - Remark on the (non)convergence of ensemble densities in dynamical systems. AB - We consider a dynamical system with state space M, a smooth, compact subset of some R(n), and evolution given by T(t), x(t)=T(t)x, x in M; T(t) is invertible and the time t may be discrete, t in Z, T(t)=T(t), or continuous, t in R. Here we show that starting with a continuous positive initial probability density rho(x,0)>0, with respect to dx, the smooth volume measure induced on M by Lebesgue measure on R(n), the expectation value of logrho(x,t), with respect to any stationary (i.e., time invariant) measure nu(dx), is linear in t, nu(logrho(x,t))=nu(logrho(x,0))+Kt. K depends only on nu and vanishes when nu is absolutely continuous with respect to dx.(c) 1998 American Institute of Physics. PMID- 12779744 TI - Entropy balance, time reversibility, and mass transport in dynamical systems. AB - We review recent results concerning entropy balance in low-dimensional dynamical systems modeling mass (or charge) transport. The key ingredient for understanding entropy balance is the coarse graining of the local phase-space density. It mimics the fact that ever refining phase-space structures caused by chaotic dynamics can only be detected up to a finite resolution. In addition, we derive a new relation for the rate of irreversible entropy production in steady states of dynamical systems: It is proportional to the average growth rate of the local phase-space density. Previous results for the entropy production in steady states of thermostated systems without density gradients and of Hamiltonian systems with density gradients are recovered. As an extension we derive the entropy balance of dissipative systems with density gradients valid at any instant of time, not only in stationary states. We also find a condition for consistency with thermodynamics. A generalized multi-Baker map is used as an illustrative example. (c) 1998 American Institute of Physics. PMID- 12779745 TI - Chaotic and fractal properties of deterministic diffusion-reaction processes. AB - We study the consequences of deterministic chaos for diffusion-controlled reaction. As an example, we analyze a diffusive-reactive deterministic multibaker and a parameter-dependent variation of it. We construct the diffusive and the reactive modes of the models as eigenstates of the Frobenius-Perron operator. The associated eigenvalues provide the dispersion relations of diffusion and reaction and, hence, they determine the reaction rate. For the simplest model we show explicitly that the reaction rate behaves as phenomenologically expected for one dimensional diffusion-controlled reaction. Under parametric variation, we find that both the diffusion coefficient and the reaction rate have fractal-like dependences on the system parameter. (c) 1998 American Institute of Physics. PMID- 12779746 TI - An analytical construction of the SRB measures for Baker-type maps. AB - For a class of dynamical systems, called the axiom-A systems, Sinai, Ruelle and Bowen showed the existence of an invariant measure (SRB measure) weakly attracting the temporal average of any initial distribution that is absolutely continuous with respect to the Lebesgue measure. Recently, the SRB measures were found to be related to the nonequilibrium stationary state distribution functions for thermostated or open systems. Inspite of the importance of these SRB measures, it is difficult to handle them analytically because they are often singular functions. In this article, for three kinds of Baker-type maps, the SRB measures are analytically constructed with the aid of a functional equation, which was proposed by de Rham in order to deal with a class of singular functions. We first briefly review the properties of singular functions including those of de Rham. Then, the Baker-type maps are described, one of which is nonconservative but time reversible, the second has a Cantor-like invariant set, and the third is a model of a simple chemical reaction R<-->I<-->P. For the second example, the cases with and without escape are considered. For the last example, we consider the reaction processes in a closed system and in an open system under a flux boundary condition. In all cases, we show that the evolution equation of the distribution functions partially integrated over the unstable direction is very similar to de Rham's functional equation and, employing this analogy, we explicitly construct the SRB measures. (c) 1998 American Institute of Physics. PMID- 12779747 TI - Bogoliubov-Born-Green-Kirkwood-Yvon hierarchy methods for sums of Lyapunov exponents for dilute gases. AB - We consider a general method for computing the sum of positive Lyapunov exponents for moderately dense gases. This method is based upon hierarchy techniques used previously to derive the generalized Boltzmann equation for the time-dependent spatial and velocity distribution functions for such systems. We extend the variables in the generalized Boltzmann equation to include a new set of quantities that describe the separation of trajectories in phase space needed for a calculation of the Lyapunov exponents. The method described here is especially suitable for calculating the sum of all of the positive Lyapunov exponents for the system, and may be applied to equilibrium as well as nonequilibrium situations. For low densities we obtain an extended Boltzmann equation, from which, under a simplifying approximation, we recover the sum of positive Lyapunov exponents for hard-disk and hard-sphere systems, obtained before by a simpler method. In addition we indicate how to improve these results by avoiding the simplifying approximation. The restriction to hard-sphere systems in d dimensions is made to keep the somewhat complicated formalism as clear as possible, but the method can be easily generalized to apply to gases of particles that interact with strong short-range forces. (c) 1998 American Institute of Physics. PMID- 12779748 TI - Lyapunov instability of two-dimensional fluids: Hard dumbbells. AB - We generalize Benettin's classical algorithm for the computation of the full Lyapunov spectrum to the case of a two-dimensional fluid composed of linear molecules modeled as hard dumbbells. Each dumbbell, two hard disks of diameter sigma with centers separated by a fixed distance d, may translate and rotate in the plane. We study the mixing between these qualitatively different degrees of freedom and its influence on the full set of Lyapunov exponents. The phase flow consists of smooth streaming interrupted by hard elastic collisions. We apply the exact collision rules for the differential offset vectors in tangent space to the computation of the Lyapunov exponents, and of time-averaged offset-vector projections into various subspaces of the phase space. For the case of a homogeneous mass distribution within a dumbbell we find that for small enough d/sigma, depending on the density, the translational part of the Lyapunov spectrum is decoupled from the rotational part and converges to the spectrum of hard disks. (c) 1998 American Institute of Physics. PMID- 12779749 TI - Entropy evolution for the Baker map. AB - Gibbs entropy is invariant for the Baker map. A Jordan basis spectral decomposition of the Baker Frobenius-Perron operator suggests that any initial density evolves to the stationary density that has maximal entropy. This entropy conundrum is resolved by considering the difference between weak and strong convergence. A binary representation is used to make these points transparent. (c) 1998 American Institute of Physics. PMID- 12779750 TI - Convergence of Hamiltonian systems to billiards. AB - We examine in detail a physically natural and general scheme for gradually deforming a Hamiltonian to its corresponding billiard, as a certain parameter k varies from one to infinity. We apply this limiting process to a class of Hamiltonians with homogeneous potential-energy functions and further investigate the extent to which the limiting billiards inherit properties from the corresponding sequences of Hamiltonians. The results are mixed. Using theorems of Yoshida for the case of two degrees of freedom, we prove a general theorem establishing the "inheritability" of stability properties of certain orbits. This result follows naturally from the convergence of the traces of appropriate monodromy matrices to the billiard analog. However, in spite of the close analogy between the concepts of integrability for Hamiltonian systems and billiards, integrability properties of Hamiltonians in a sequence are not necessarily inherited by the limiting billiard, as we show by example. In addition to rigorous results, we include numerical examples of certain interesting cases, along with computer simulations. (c) 1998 American Institute of Physics. PMID- 12779751 TI - Order and chaos in the planar isosceles three-body problem. AB - The planar isosceles three-body problem has been reduced to a two-dimensional area preserving Poincare map f. Using certain symmetry properties of the underlying differential equations and numerical integration, we offer a global description of f in the case of three equal masses. This description, which is based on the mapping of areas, immediately leads to the existence of various types of motion such as capture-escape, permanent capture, ejection-collision, etc., and their corresponding measures in the map domain. Moreover, this technique readily allows one to distinguish between so-called "fast" and "chaotic" scattering. Although capture-escape is the subset with the highest measure, there exist two important distinct invariant subsets under f where the solutions neither are captured nor lead to escape. The first set is a Cantor set which has zero measure and it is the outcome of the fact that f acts similar to the Smale horseshoe map in part of the domain. On this subset the action of f is chaotic. The second subset is an invariant region with positive measure surrounding an elliptic fixed point. In this region f acts essentially as a perturbed twist mapping where the iterates of f for the points in a large subset move on invariant curves in an orderly manner. In an appendix we cast our results in the framework of the widely studied isosceles triple collision manifold. (c) 1998 American Institute of Physics. PMID- 12779752 TI - On the evidence of deterministic chaos in ECG: Surrogate and predictability analysis. AB - The question whether the human cardiac system is chaotic or not has been an open one. Recent results in chaos theory have shown that the usual methods, such as saturation of correlation dimension D(2) or the existence of positive Lyapunov exponent, alone do not provide sufficient evidence to confirm the presence of deterministic chaos in an experimental system. The results of surrogate data analysis together with the short-term prediction analysis can be used to check whether a given time series is consistent with the hypothesis of deterministic chaos. In this work nonlinear dynamical tools such as surrogate data analysis, short-term prediction, saturation of D(2) and positive Lyapunov exponent have been applied to measured ECG data for several normal and pathological cases. The pathology presently studied are PVC (Premature Ventricular Contraction), VTA (Ventricular Tachy Arrhythmia), AV (Atrio-Ventricular) block and VF (Ventricular Fibrillation). While these results do not prove that ECG time series is definitely chaotic, they are found to be consistent with the hypothesis of chaotic dynamics. (c) 1998 American Institute of Physics. PMID- 12779753 TI - Chaos in a dynamic model of traffic flows in an origin-destination network. AB - In this paper we investigate the dynamic behavior of road traffic flows in an area represented by an origin-destination (O-D) network. Probably the most widely used model for estimating the distribution of O-D flows is the gravity model, [J. de D. Ortuzar and L. G. Willumsen, Modelling Transport (Wiley, New York, 1990)] which originated from an analogy with Newton's gravitational law. The conventional gravity model, however, is static. The investigation in this paper is based on a dynamic version of the gravity model proposed by Dendrinos and Sonis by modifying the conventional gravity model [D. S. Dendrinos and M. Sonis, Chaos and Social-Spatial Dynamics (Springer-Verlag, Berlin, 1990)]. The dynamic model describes the variations of O-D flows over discrete-time periods, such as each day, each week, and so on. It is shown that when the dimension of the system is one or two, the O-D flow pattern either approaches an equilibrium or oscillates. When the dimension is higher, the behavior found in the model includes equilibria, oscillations, periodic doubling, and chaos. Chaotic attractors are characterized by (positive) Liapunov exponents and fractal dimensions.(c) 1998 American Institute of Physics. PMID- 12779754 TI - Chaos suppression in gas-solid fluidization. AB - Fluidization in granular materials occurs primarily as a result of a dynamic balance between gravitational forces and forces resulting from the flow of a fluid through a bed of discrete particles. For systems where the fluidizing medium and the particles have significantly different densities, density wave instabilities create local pockets of very high void fraction termed bubbles. The fluidization regime is termed the bubbling regime. Such a system is appropriately termed a self-excited nonlinear system. The present study examines chaos suppression resulting from an opposing oscillatory flow in gas-solid fluidization. Time series data representing local, instantaneous pressure were acquired at the surface of a horizontal cylinder submerged in a bubbling fluidized bed. The particles had a weight mean diameter of 345 &mgr;m and a narrow size distribution. The state of fluidization corresponded to the bubbling regime and total air flow rates employed in the present study ranged from 10% to 40% greater than that required for minimum fluidization. The behavior of time varying local pressure in fluidized beds in the absence of a secondary flow is consistent with deterministic chaos. Kolmogorov entropy estimates from local, instantaneous pressure suggest that the degree of chaotic behavior can be substantially suppressed by the presence of an opposing, oscillatory secondary flow. Pressure signals clearly show a "phase-locking" phenomenon coincident with the imposed frequency. In the present study, the greatest degree of suppression occurred for operating conditions with low primary and secondary flow rates, and a secondary flow oscillation frequency of 15 Hz. (c) 1998 American Institute of Physics. PMID- 12779755 TI - The effect of convection on a propagating front with a liquid product: Comparison of theory and experiments. AB - This work is devoted to the investigation of propagating polymerization fronts converting a liquid monomer into a liquid polymer. We consider a simplified mathematical model which consists of the heat equation and equation for the depth of conversion for one-step chemical reaction and of the Navier-Stokes equations under the Boussinesq approximation. We fulfill the linear stability analysis of the stationary propagating front and find conditions of convective and thermal instabilities. We show that convection can occur not only for ascending fronts but also for descending fronts. Though in the latter case the exothermic chemical reaction heats the cold monomer from above, the instability appears and can be explained by the interaction of chemical reaction with hydrodynamics. Hydrodynamics changes also conditions of the thermal instability. The front propagating upwards becomes less stable than without convection, the front propagating downwards more stable. The theoretical results are compared with experiments. The experimentally measured stability boundary for polymerization of benzyl acrylate in dimethyl formamide is well approximated by the theoretical stability boundary. (c) 1998 American Institute of Physics. PMID- 12779756 TI - Overview: The constructive role of noise in fluctuation driven transport and stochastic resonance. AB - Random noise is typically thought of as the enemy of order rather than as a constructive influence. Recent work has shown however that under certain circumstances, noise and Brownian motion can facilitate transmission of information via a mechanism know as stochastic resonance, and help systems use chemical energy and nonequilibrium fluctuations to drive directed motion via fluctuation driven transport. In this focus issue we have collected several articles that capture the flavor of these developing fields and point the way to new directions for research. (c) 1998 American Institute of Physics. PMID- 12779757 TI - Minireview of stochastic resonance. AB - We present an introductory overview of the subject of stochastic resonance. As researchers' interest in the phenomenon has spread from physics to biology, new questions both fundamental and practical have emerged. After reviewing some key aspects of the subject, we describe a promising candidate for exploring the possible beneficial effects of random noise in sensory transduction. (c) 1998 American Institute of Physics. PMID- 12779758 TI - Barrier fluctuations and stochastic resonance in membrane transport. AB - The role of barrier fluctuations in membrane enzymatic processes, in particular in the active transport of ions through cell membranes, is examined. For enzymes embedded in the cell membrane the role of the barrier height (activation energy) is played by the membrane electric potential. This barrier height can be modulated either by internal fluctuations or by external electrical fields, either random or periodic. Existing experimental data on active transport of Na(+) and Rb(+) in human erythrocytes (catalyzed by Na(+)-K(+)-ATPase) can be interpreted as evidence of stochastic resonance between the external ac field and the fluctuations of the membrane potential. The obtained results suggest that the significant part of these fluctuations is supplied by the stimulated action of neighbor voltage-gated ionic channels. This supports the idea that intrinsic noise plays a constructive role in one of most important and most frequent biophysical processs, viz. ion transmission through cell membranes. Means of further experimental verification of this conjecture are proposed. (c) 1998 American Institute of Physics. PMID- 12779759 TI - Stochastic resonance in thermally activated reactions: Application to biological ion channels. AB - At the molecular level many thermally activated reactions can be viewed as Poisson trains of events whose instantaneous rates are defined by the reaction activation barrier height and an effective collision frequency. When the barrier height depends on an external parameter, variation in this parameter induces variation in the event rate. Extending our previous work, we offer a detailed theoretical analysis of signal transduction properties of these reactions considering the external parameter as an input signal and the train of resulting events as an output signal. The addition of noise to the system input facilitates signal transduction in two ways. First, for a linear relationship between the barrier height and the external parameter the output signal power grows exponentially with the mean square fluctuation of the noise. Second, for noise of a sufficiently high bandwidth, its addition increases output signal quality measured as the signal-to-noise ratio (SNR). The output SNR reaches a maximum at optimal noise intensity defined by the reaction sensitivity to the external parameter, reaction initial rate, and the noise bandwidth. We apply this theory to ion channels of excitable biological membranes. Based on classical results of Hodgkin and Huxley we show that open/closed transitions of voltage-gated ion channels can be treated as thermally activated reactions whose activation barriers change linearly with applied transmembrane voltage. As an experimental example we discuss our recent results obtained with polypeptide alamethicin incorporated into planar lipid bilayers.(c) 1998 American Institute of Physics. PMID- 12779760 TI - Noise sustained waves in subexcitable media: From chemical waves to brain waves. AB - We discuss a novel type of spatiotemporal pattern that can be observed in subexcitable media when coupled to a thermal environment. These patterns have been recently observed in several different types of systems: a subexcitable photosensitive Belousov-Zhabotinsky reaction, hippocampal slices of rat brains, and astrocyte syncytium. In this paper, we introduce the basic concepts of subexcitable media, describe recent experimental observations in chemistry and neurophysiology, and put these observation into context with computer simulations. (c) 1998 American Institute of Physics. PMID- 12779761 TI - Detection of weak electric fields by sharks, rays, and skates. AB - The elasmobranchs-sharks, rays, and skates-can detect very weak electric fields in their aqueous environment through a complex sensory system, the ampullae of Lorenzini. The ampullae are conducting tubes that connect the surface of the animal to its interior. In the presence of an electric field, the potential of the surface of the animal will differ from that of the interior and that potential is applied across the apical membrane of the special sensory cells that line the ampullae. The firing rate of the afferent neurons that transmit signals from the ampullae has been shown to vary with that potential. We show that those firing rates can be described quantitatively in terms of synchronous firing of the sensory cells that feed the neurons. We demonstrate that such synchronism follows naturally from a hypothetical weak cell-to-cell interaction that results in a self-organization of the sensory cells. Moreover, the pulse rates of those cells-and the neurons that service the cells-can be expected to vary with the imposed electric fields in accord with measured values through actions of voltage gated transmembrane proteins in the apical sector of the cell membranes that admit Ca(++) ions. We also present a more conjectural model of signal processing at the neuron level that could exploit small differences in firing rates of nerve fibers servicing different ampullae to send an unambiguous signal to the central nervous system of the animal. (c) 1998 American Institute of Physics. PMID- 12779762 TI - Stochastic resonance in mammalian neuronal networks. AB - We present stochastic resonance observed in the dynamics of neuronal networks from mammalian brain. Both sinusoidal signals and random noise were superimposed into an applied electric field. As the amplitude of the noise component was increased, an optimization (increase then decrease) in the signal-to-noise ratio of the network response to the sinusoidal signal was observed. The relationship between the measures used to characterize the dynamics is discussed. Finally, a computational model of these neuronal networks that includes the neuronal interactions with the electric field is presented to illustrate the physics behind the essential features of the experiment. (c) 1998 American Institute of Physics. PMID- 12779763 TI - Using electrical noise to enhance the ability of humans to detect subthreshold mechanical cutaneous stimuli. AB - Stochastic resonance (SR) is a phenomenon wherein the response of a nonlinear system to a weak input signal is optimized by the presence of a particular, nonzero level of noise. Our objective was to demonstrate cross-modality SR in human sensory perception. Specifically, we were interested in testing the hypothesis that the ability of an individual to detect a subthreshold mechanical cutaneous stimulus can be significantly enhanced by introducing a particular level of electrical noise. Psychophysical experiments were performed on 11 healthy subjects. The protocol consisted of the presentation of: (a) a subthreshold mechanical stimulus plus electrical noise, or (b) no mechanical stimulus plus electrical noise. The intensity of the electrical noise was varied between trials. Each subject's ability to identify correctly the presence of the mechanical stimulus was determined as a function of the noise intensity. In 9 of the 11 subjects, the introduction of a particular level of electrical noise significantly enhanced the subject's ability to detect the subthreshold mechanical cutaneous stimulus. In 2 of the 11 subjects, the introduction of electrical noise did not significantly change the subject's ability to detect the mechanical stimulus. These findings indicate that input electrical noise can serve as a negative masker for subthreshold mechanical tactile stimuli, i.e., electrical noise can increase the detectability of weak mechanical signals. Thus, for SR-type effects to be observed in human sensory perception, the noise and stimulus need not be of the same modality. From a bioengineering and clinical standpoint, this work suggests that an electrical noise-based technique could be used to improve tactile sensation in humans when the mechanical stimulus is around or below threshold. (c) 1998 American Institute of Physics. PMID- 12779764 TI - Stochastic resonance in coupled nonlinear dynamic elements. AB - We investigate the response of a linear chain of diffusively coupled diode resonators under the influence of thermal noise. We also examine the connection between spatiotemporal stochastic resonance and the presence of kink-antikink pairs in the array. The interplay of nucleation rates and kink speeds is briefly addressed. The experimental results are supplemented with simulations on a coupled map lattice. We furthermore present analytical results for the synchronization and signal processing properties of a Phi(4) field theory and explore the effects of various forms of nonlinear coupling. (c) 1998 American Institute of Physics. PMID- 12779765 TI - Enhancing aperiodic stochastic resonance through noise modulation. AB - We show that the conventional stochastic resonance (SR) effect for aperiodic signals in a model neuron can be enhanced by modulating the intensity of the input noise (which could be introduced artificially in bioengineering applications) with either the input signal or the unit's output rate signal. We analyze SR enhancement theoretically and numerically. We discuss how this work provides the theoretical foundation for the development of an optimal noise-based technique for enhancing sensory function. (c) 1998 American Institute of Physics. PMID- 12779766 TI - Communicating with noise: How chaos and noise combine to generate secure encryption keys. AB - An approach for the secure transmission of encrypted messages using chaos and noise is presented in this paper. The method is based on the synchronization of certain types of chaotic oscillators in response to a common noise input. This allows two distant oscillators to generate identical output which can be used as a key for encryption and decryption of a message signal. The noiselike synchronizing input-which contains no message information-is communicated to identical oscillators in the transmitter and the receiver over a public channel. The encrypted message is also sent over a public channel, while the key is never transmitted at all. The chaotic nature of the oscillators which generate the key and the randomness of the signal driving the process combine to make the recovery of the key by an eavesdropper extremely difficult. We evaluate system performance with respect to security and robustness and show that a robust and secure system can be obtained. (c) 1998 American Institute of Physics. PMID- 12779767 TI - Quantum features of Brownian motors and stochastic resonance. AB - We investigate quantum Brownian motion sustained transport in both, adiabatically rocked ratchet systems and quantum stochastic resonance (QSR). Above a characteristic crossover temperature T(0) tunneling events are rare; yet they can considerably enhance the quantum-noise-driven particle current and the amplification of signal output in comparison to their classical counterparts. Below T(0) tunneling prevails, thus yielding characteristic novel quantum transport phenomena. For example, upon approaching T=0 the quantum current in Brownian motors exhibits a tunneling-induced reversal, and tends to a finite limit, while the classical result approaches zero without such a change of sign. As a consequence, similar current inversions generated by quantum effects follow upon variation of the particle mass or of its friction coefficient. Likewise, in this latter regime of very low temperatures the tunneling dynamics becomes increasingly coherent, thus suppressing the semiclassically predicted QSR. Moreover, nonadiabatic driving may cause driving-induced coherences and quantized resonant transitions with no classical analog. (c) 1998 American Institute of Physics. PMID- 12779768 TI - Constructive role of noise: Fast fluctuation asymptotics of transport in stochastic ratchets. AB - The constructive role of random fluctuations is studied in the context of transport in stochastic ratchets. We discuss the interplay of independent white (thermal) and discrete (external) noises and their generation of transport in anisotropic potentials. The constructive cooperation of such fluctuations is most apparent in the asymptotic limit of fast discrete-valued noise, a limit which presents some interesting mathematical features. We describe the asymptotic analysis of the current in the limit of fast external noise, pointing out the strong qualitative dependence of the current on the interplay of the independent noise sources and its surprising sensitivity to the regularity of the underlying anisotropic ratchet potential. (c) 1998 American Institute of Physics. PMID- 12779769 TI - Dielectrophoretic ratchets. AB - We have experimentally applied some concepts of "force-free" motion to micron size particles (latex beads). The coupling of dissipation and local spatial asymmetry of the potential experienced by the beads can put them into motion. The potentials used in these experiments are of dielectrophoretic nature. To that end, electrodes of particular shapes were used in order to submit the considered suspensions to inhomogeneous ac electric fields. Two regimes were explored: i-the Brownian ratchet case in which a Brownian particle is successively trapped in a factory roof-like potential and left free to diffuse. ii-the shifted ratchets case in which two potentials exhibiting similar characteristics are applied successively, one of them being shifted by a fraction of their common period relatively to the other. In both cases, a good agreement with the theoretical predictions was observed. In particular, particles of different sizes were characterized by different macroscopic velocities leading to the prospect of promising separation techniques. (c) 1998 American Institute of Physics. PMID- 12779770 TI - Collective transport in locally asymmetric periodic structures. AB - In this paper we give an overview of the cooperative effects in fluctuation driven transport arising from the interaction of a large number of particles. (i) First, we study a model with finite-sized, overdamped Brownian particles interacting via hard-core repulsion. Computer simulations and theoretical calculations reveal a number of novel cooperative transport phenomena in this system, including the reversal of direction of the net current as the particle density is increased, and a very strong and complex dependence of the average velocity on both the size and the average distance of the particles. (ii) Next, we consider the cooperation of a collection of motors rigidly attached to a backbone. This system possesses dynamical phase transition allowing spontaneous directed motion even if the system is spatially symmetric. (iii) Finally, we report on an experimental investigation exploring the horizontal transport of granular particles in a vertically vibrated system whose base has a sawtooth shaped profile. The resulting material flow exhibits complex collective behavior, both as a function of the number of layers of particles and the driving frequency; in particular, under certain conditions, increasing the layer thickness leads to a reversal of the current, while the onset of transport as a function of frequency occurs gradually in a manner reminiscent of a phase transition. (c) 1998 American Institute of Physics. PMID- 12779771 TI - Exponential asymptotic expansions and approximations of the unstable and stable manifolds of singularly perturbed systems with the Henon map as an example. AB - The subject of this paper is the construction of the exponential asymptotic expansions of the unstable and stable manifolds of the area-preserving Henon map. The approach that is taken enables one to capture the exponentially small effects that result from what is known as the Stokes phenomenon in the analytic theory of equations with irregular singular points. The exponential asymptotic expansions were then used to obtain explicit functional approximations for the stable and unstable manifolds. These approximations are compared with numerical simulations and the agreement is excellent. Several of the main results of the paper have been previously announced in A. Tovbis, M. Tsuchiya, and C. Jaffe ["Chaos integrability transition in nonlinear dynamical systems: exponential asymptotic approach," Differential Equations and Applications to Biology and to Industry, edited by M. Martelli, K. Cooke, E. Cumberbatch, B. Tang, and H. Thieme (World Scientific, Singapore, 1996), pp. 495-507, and A. Tovbis, M. Tsuchiya, and C. Jaffe, "Exponential asymptotic expansions and approximations of the unstable and stable manifolds of the Henon map," preprint, 1994]. (c) 1998 American Institute of Physics. PMID- 12779772 TI - A spectroscopical measure for the exploration of phase space. AB - Many nonintegrable systems have eigenstates that typically require numerous basis states to represent them. We develop a criterion to judge the extent to which phase space is explored by the spectrum of such a Hamiltonian. Our criterion uses the eigenvalues rather than the eigenfunctions and is based on identifying a direct relation between the intensity of Shnirelman's peak and the localization length. We illustrate our procedure by applying it to the spectrum of two prototypical nonintegrable systems, the kicked rotor and the kicked top. (c) 1998 American Institute of Physics. PMID- 12779773 TI - Data compression and information retrieval via symbolization. AB - Converting a continuous signal into a multisymbol stream is a simple method of data compression which preserves much of the dynamical information present in the original signal. The retrieval of selected types of information from symbolic data involves binary operations and is therefore optimal for digital computers. For example, correlation time scales can be easily recovered, even at high noise levels, by varying the time delay for symbolization. Also, the presence of periodicity in the signal can be reliably detected even if it is weak and masked by a dominant chaotic/stochastic background. (c) 1998 American Institute of Physics. PMID- 12779774 TI - Stages of chaotic synchronization. AB - In an experimental investigation of the response of a chaotic system to a chaotic driving force, we have observed synchronization of chaos of the response system in the forms of generalized synchronization, phase synchronization, and lag synchronization to the driving signal. In this paper we compare the features of these forms of synchronized chaos and study their relations and physical origins. We found that different forms of chaotic synchronization could be interpreted as different stages of nonlinear interaction between the coupled chaotic systems. (c) 1998 American Institute of Physics. PMID- 12779775 TI - Communication with chemical chaos in the presence of noise. AB - We use control of chaos to encode information into the oscillations of the Belousov-Zhabotinsky reaction. An arbitrary binary message is encoded by forcing the chaotic oscillations to follow a specified trajectory. The information manipulating control requires only small perturbations to vary the binary message. In this paper we extend our recent theoretical work [Bollt and Dolnik, Phys. Rev. E 64, 1196 (1990)] by introducing a new and simplified encoding technique which can be utilized in the presence of experimental noise. We numerically and theoretically study several practical aspects of controlling symbol dynamics including: modeling noisy time-series, learning underlying symbol dynamics, and evaluation of derivatives for control by observing system responses to an intelligent and deliberate sequence of input parameter variations. All of the modeling techniques incorporated here are ultimately designed to learn and control symbol dynamics of experimental data known only as an observed time series; the simulation assumes no global model. We find that noise affects reliability of encoding information and may cause coding errors. But, if the level of noise is confined to relatively small values, which are achievable in experiments, the control mechanism is robust to the noise. Thus we can still produce a desired symbolic code. However, scarce errors in encoding may occur due to rare but large fluctuations. These errors may be corrected during the decoding process by a variation of the filtering technique suggested by Rosa et al. [Phys. Rev. Lett. 78, 1247 (1997)]. (c) 1998 American Institute of Physics. PMID- 12779776 TI - Internal symmetries of cellular automata via their polynomial representation. AB - A polynomial representation of elementary cellular automata (ECA) is used to give a complete characterization of the local internal symmetries of all ECA. It is also shown that the polynomial representation is a natural choice for the study of local internal transformations of all cellular automata with two symbols. This is achieved by proving that local internal transformations are simply expressed in this representation as sums of polynomials. (c) 1998 American Institute of Physics. PMID- 12779777 TI - How organisms do the right thing: The attractor hypothesis. AB - Neo-Darwinian theory is highly successful at explaining the emergence of adaptive traits over successive generations. However, there are reasons to doubt its efficacy in explaining the observed, impressively detailed adaptive responses of organisms to day-to-day changes in their surroundings. Also, the theory lacks a clear mechanism to account for both plasticity and canalization. In effect, there is a growing sentiment that the neo-Darwinian paradigm is incomplete, that something more than genetic structure, mutation, genetic drift, and the action of natural selection is required to explain organismal behavior. In this paper we extend the view of organisms as complex self-organizing entities by arguing that basic physical laws, coupled with the acquisitive nature of organisms, makes adaptation all but tautological. That is, much adaptation is an unavoidable emergent property of organisms' complexity and, to some a significant degree, occurs quite independently of genomic changes wrought by natural selection. For reasons that will become obvious, we refer to this assertion as the attractor hypothesis. The arguments also clarify the concept of "adaptation." Adaptation across generations, by natural selection, equates to the (game theoretic) maximization of fitness (the success with which one individual produces more individuals), while self-organizing based adaptation, within generations, equates to energetic efficiency and the matching of intake and biosynthesis to need. Finally, we discuss implications of the attractor hypothesis for a wide variety of genetical and physiological phenomena, including genetic architecture, directed mutation, genetic imprinting, paramutation, hormesis, plasticity, optimality theory, genotype-phenotype linkage and puncuated equilibrium, and present suggestions for tests of the hypothesis. (c) 1998 American Institute of Physics. PMID- 12779778 TI - Dynamical properties of a ferroelectric capacitor observed through nonlinear time series analysis. AB - By data analysis the ordinary differential equation for the description of an experimental electric resonance circuit with nonlinear capacitor is derived. Triglycine sulfate (TGS) was used as nonlinear dielectric material. This is the most thoroughly investigated ferroelectric with a second order phase transition. Its static dielectric small signal behavior is well described in the framework of the Landau theory, yielding a Duffing-type ordinary differential equation as a model equation of the circuit. Data analysis allows us to check carefully the validity of this model and to determine required corrections of this simplified equation. (c) 1998 American Institute of Physics. PMID- 12779779 TI - Ordering of the Mandelbrot-like set of the exponential map. AB - We graphically study the Mandelbrot-like set of the complex exponential family of maps E(lambda)(z)=lambdae(z), which we call the Baker-Rippon-Devaney (BRD) set. We observe that the period of every hyperbolic component can be deduced with the naked eye by using two simple rules. (c) 1998 American Institute of Physics. PMID- 12779780 TI - Spectral decomposition of the tent map with varying height. AB - The generalized spectral decomposition of the Frobenius-Perron operator of the tent map with varying height is determined at the band-splitting points. The decomposition includes both decay onto the attracting set and the approach to the asymptotically periodic state on the attractor. Explicit compact expressions for the polynomial eigenstates are obtained using algebraic techniques. (c) 1998 American Institute of Physics. PMID- 12779782 TI - Aperiodic stochastic resonance in chaotic maps. AB - It is shown by means of numerical simulations that aperiodic stochastic resonance occurs in chaotic one-dimensional maps with various kinds of intermittency. The effect appears in the absence of external noise, as the system control parameter is varied. In the case of input signals slowly varying in time the analytic treatment, using the adiabatic approximation based on the expressions for the mean laminar phase duration, yields the input-output covariance function comparable with numerical results. (c) 1998 American Institute of Physics. PMID- 12779781 TI - Near threshold anomalous transport in the standard map. AB - Anomalous transport is investigated near threshold in the standard map. Very long time flights, and a large anomaly in the transport, are shown to be associated with a new form of multi-island structures causing orbit sticking. The phase space structure of these traps, and the exponents of the characteristic long time tails associated with them are determined. In general these structures are very complex, but some cases, consisting of layers of islands, allow simple modeling. (c) 1998 American Institute of Physics. PMID- 12779783 TI - Nonlinear noise reduction through Monte Carlo sampling. AB - We consider the problem of nonlinear noise reduction within the framework of Bayesian Theory. This enables us to place appropriate weights on the measurement and dynamic errors and thereby avoid over cleaning the data. Using a Metropolis Hastings sampler, we are able to achieve robust noise reduction without the introduction of ad hoc parameters but at the expense of higher computational complexity. Such an algorithm should also allow us to explore the potential and limitations of other noise reduction methods. (c) 1998 American Institute of Physics. PMID- 12779784 TI - Stabilizing unstable steady states using extended time-delay autosynchronization. AB - We describe a method for stabilizing unstable steady states in nonlinear dynamical systems using a form of extended time-delay autosynchronization. Specifically, stabilization is achieved by applying a feedback signal generated by high-pass-filtering in real time the dynamical state of the system to an accessible system parameter or variables. Our technique is easy to implement, does not require knowledge of the unstable steady state coordinates in phase space, automatically tracks changes in the system parameters, and is more robust to broadband noise than previous schemes. We demonstrate the controller's efficacy by stabilizing unstable steady states in an electronic circuit exhibiting low-dimensional temporal chaos. The simplicity and robustness of the scheme suggests that it is ideally suited for stabilizing unstable steady states in ultra-high-speed systems. (c) 1998 American Institute of Physics. PMID- 12779785 TI - A novel nonlinear car-following model. AB - The mathematical models used to describe the dynamical behavior of a group of road vehicles traveling in a single lane without overtaking are known as car following models. These models are widely used in many commercially available microscopic traffic simulation software packages. They attempt to mimic the interactions between individual vehicles that are traveling sufficiently close together for the behavior of each vehicle to be dependent upon the motion of the vehicle immediately in front. In this paper we modify the traditional car following model by adding a new nonlinear term to take account of the driver attempting to achieve a certain desired intervehicle separation distance as well as the traditional aim of matching the velocity of the vehicle ahead. Numerical solution of the resulting coupled system of nonlinear differential equations is used to analyze the stability of the equilibrium solution to a periodic perturbation. For certain parameter values chaotic oscillations are generated, consisting of a broad spectrum of frequency components. Such chaotic motion produces extremely complicated dynamical behavior that has an inherent lack of predictability associated with it. The results of simulating over a range of parameter values are presented and, where it is present, the degree of chaos is estimated. (c) 1998 American Institute of Physics. PMID- 12779786 TI - Using chaos to generate variations on movement sequences. AB - We describe a method for introducing variations into predefined motion sequences using a chaotic symbol-sequence reordering technique. A progression of symbols representing the body positions in a dance piece, martial arts form, or other motion sequence is mapped onto a chaotic trajectory, establishing a symbolic dynamics that links the movement sequence and the attractor structure. A variation on the original piece is created by generating a trajectory with slightly different initial conditions, inverting the mapping, and using special corpus-based graph-theoretic interpolation schemes to smooth any abrupt transitions. Sensitive dependence guarantees that the variation is different from the original; the attractor structure and the symbolic dynamics guarantee that the two resemble one another in both aesthetic and mathematical senses. (c) 1998 American Institute of Physics. PMID- 12779787 TI - Chaotic evolution of arms races. AB - A new set of model equations is proposed to describe the evolution of the arms race, by extending Richardson's model with special emphases that (1) power dependent defensive reaction or historical enmity could be a motive force to promote armaments, (2) a deterrent would suppress the growth of armaments, and (3) the defense reaction of one nation against the other nation depends nonlinearly on the difference in armaments between two. The set of equations is numerically solved to exhibit stationary, periodic, and chaotic behavior depending on the combinations of parameters involved. The chaotic evolution is realized when the economic situation of each country involved in the arms race is quite different, which is often observed in the real world. (c) 1998 American Institute of Physics. PMID- 12779788 TI - Synchronization of cellular automaton pairs. AB - The phenomenon of synchronization in pairs of cellular automata coupled in a driver-replica mode is studied. Necessary and sufficient conditions for synchronization in linear cellular automaton pairs are given. The couplings that make a pair synchronize are determined for all linear elementary cellular automata. (c) 1998 American Institute of Physics. PMID- 12779789 TI - A cryptosystem based on cellular automata. AB - Cryptosystems for binary information are based on two primitives: an indexed family of permutations of binary words and a generator of pseudorandom sequences of indices. A very efficient implementation of the primitives is constructed using the phenomenon of synchronization in cellular automata. (c) 1998 American Institute of Physics. PMID- 12779790 TI - (Global and local) fluctuations of phase space contraction in deterministic stationary nonequilibrium. AB - We studied numerically the validity of the fluctuation relation introduced in Evans et al. [Phys. Rev. Lett. 71, 2401-2404 (1993)] and proved under suitable conditions by Gallavotti and Cohen [J. Stat. Phys. 80, 931-970 (1995)] for a two dimensional system of particles maintained in a steady shear flow by Maxwell demon boundary conditions [Chernov and Lebowitz, J. Stat. Phys. 86, 953-990 (1997)]. The theorem was found to hold if one considers the total phase space contraction sigma occurring at collisions with both walls: sigma=sigma( upward arrow )+sigma( downward arrow ). An attempt to extend it to more local quantities sigma( upward arrow ) and sigma( downward arrow ), corresponding to the collisions with the top or bottom wall only, gave negative results. The time decay of the correlations in sigma( upward arrow, downward arrow ) was very slow compared to that of sigma. (c) 1998 American Institute of Physics. PMID- 12779791 TI - Systematic derivation of amplitude equations and normal forms for dynamical systems. AB - We present a systematic approach to deriving normal forms and related amplitude equations for flows and discrete dynamics on the center manifold of a dynamical system at local bifurcations and unfoldings of these. We derive a general, explicit recurrence relation that completely determines the amplitude equation and the associated transformation from amplitudes to physical space. At any order, the relation provides explicit expressions for all the nonvanishing coefficients of the amplitude equation together with straightforward linear equations for the coefficients of the transformation. The recurrence relation therefore provides all the machinery needed to solve a given physical problem in physical terms through an amplitude equation. The new result applies to any local bifurcation of a flow or map for which all the critical eigenvalues are semisimple (i.e., have Riesz index unity). The method is an efficient and rigorous alternative to more intuitive approaches in terms of multiple time scales. We illustrate the use of the method by deriving amplitude equations and associated transformations for the most common simple bifurcations in flows and iterated maps. The results are expressed in tables in a form that can be immediately applied to specific problems. (c) 1998 American Institute of Physics. PMID- 12779792 TI - Counting unstable periodic orbits in noisy chaotic systems: A scaling relation connecting experiment with theory. AB - The experimental detection of unstable periodic orbits in dynamical systems, especially those which yield short, noisy or nonstationary data sets, is a current topic of interest in many research areas. Unfortunately, for such data sets, only a few of the lowest order periods can be detected with quantifiable statistical accuracy. The primary observable is the number of encounters the general trajectory has with a particular orbit. Here we show that, in the limit of large period, this quantity scales exponentially with the period, and that this scaling is robust to dynamical noise. (c) 1998 American Institute of Physics. PMID- 12779793 TI - Recurrence plots of experimental data: To embed or not to embed? AB - A recurrence plot is a visualization tool for analyzing experimental data. These plots often reveal correlations in the data that are not easily detected in the original time series. Existing recurrence plot analysis techniques, which are primarily application oriented and completely quantitative, require that the time series data first be embedded in a high-dimensional space, where the embedding dimension d(E) is dictated by the dimension d of the data set, with d(E)>/=2d+1. One such set of recurrence plot analysis tools, recurrence quantification analysis, is particularly useful in finding locations in the data where the underlying dynamics change. We have found that for certain low-dimensional systems the same results can be obtained with no embedding. (c) 1998 American Institute of Physics. PMID- 12779794 TI - Formation and evolution of scroll waves in photosensitive excitable media. AB - Experimental and computational studies of the formation and evolution of scroll waves in three-dimensional excitable media are presented. Scroll waves are initiated in the photosensitive Belousov-Zhabotinsky reaction by perturbing traveling waves transverse to their direction of propagation. Scroll rings are generated by perturbing circular waves, which expand or contract depending on the strength of an imposed excitability gradient and its direction relative to the rotational direction of the scroll wave. (c) 1998 American Institute of Physics. PMID- 12779795 TI - Erratum: "Vortex dynamics in three-dimensional continuous myocardium with fiber rotation: Filament instability and fibrillation" [Chaos 8, 20-47 (1998)]. PMID- 12779796 TI - Editorial: Nonlinear science in chemical engineering. PMID- 12779797 TI - On some dynamical diagrams of chemical reaction engineering. AB - A brief historical survey of some of the influential types of diagrams that have been used in chemical reaction engineering is given. These include the phase plane, the simple autocatalytic diagram, and the stroboscopic phase plane. (c) 1999 American Institute of Physics. PMID- 12779798 TI - Bifurcation analysis of chemical reactors and reacting flows. AB - In this work we review the local bifurcation techniques for analyzing and classifying the steady-state and dynamic behavior of chemical reactor models described by partial differential equations (PDEs). First, we summarize the formulas for determining the derivatives of the branching equation and the coefficients in the amplitude equations for the most common singularities. We also illustrate the procedure for the numerical computation of these coefficients. Next, the application of these local results to various reactor models described by PDEs is discussed. Specifically, we review the recent literature on the bifurcation features of convection-reaction and convection diffusion-reaction models in one and more spatial dimensions, with emphasis on the features introduced due to coupling between the flow, heat and mass diffusion and chemical reaction. Finally, we illustrate the use of dynamical systems concepts in developing low dimensional (effective or pseudohomogeneous) models of reactors and reacting flows, and discuss some problems of current interest. (c) 1999 American Institute of Physics. PMID- 12779799 TI - Temperature patterns on a hollow cylindrical catalytic pellet. AB - The atmospheric oxidation of a mixture containing 6 vol % carbon monoxide was carried out on a hollow cylindrical catalytic pellet. The catalyst was held in a conical reactor which enabled simultaneous measurement of the temperature patterns on the top and side of the pellet by an IR imager. Upon a decrease in the reactor temperature the fully ignited, high temperature state of the pellet is transformed to a nonuniform one with temperature fronts separating high and low temperature regions. The transition and the resulting states are rather intricate and are strongly influenced by the nonuniformity of the catalyst and the transport to and from it, as well as the global coupling, which stabilizes temperature fronts and patterns, which would not exist in its absence. Intricate pulse splitting and extinction were observed both on the top and the side of the pellet. Highly irregular motions and conversions were obtained following a decrease in the reactor temperature. (c) 1999 American Institute of Physics. PMID- 12779800 TI - Chaotic patterns in a coupled oscillator-excitator biochemical cell system. AB - In this paper we examine dynamical modes resulting from diffusion-like interaction of two model biochemical cells. Kinetics in each of the cells is given by the ICC model of calcium ions in the cytosol. Constraints for one of the cells are set so that it is excitable. One of the constraints in the other cell - a fraction of activated cell surface receptors-is varied so that the dynamics in the cell is either excitable or oscillatory or a stable focus. The cells are interacting via mass transfer and dynamics of the coupled system are studied as two parameters are varied-the fraction of activated receptors and the coupling strength. We find that (i) the excitator-excitator interaction does not lead to oscillatory patterns, (ii) the oscillator-excitator interaction leads to alternating phase-locked periodic and quasiperiodic regimes, well known from oscillator-oscillator interactions; torus breaking bifurcation generates chaos when the coupling strength is in an intermediate range, (iii) the focus-excitator interaction generates compound oscillations arranged as period adding sequences alternating with chaotic windows; the transition to chaos is accompanied by period doublings and folding of branches of periodic orbits and is associated with a Shilnikov homoclinic orbit. The nature of spontaneous self-organized oscillations in the focus-excitator range is discussed. (c) 1999 American Institute of Physics. PMID- 12779801 TI - Ripening of surface phases coupled with oscillatory dynamics and self-induced spatial chaos through surface roughening. AB - Some pattern formation processes on single-crystal catalytic surfaces involve transitions between alternative surface phases coupled with oscillatory reaction dynamics. We describe a two-tier symmetry-breaking model of this process, based on nanoscale boundary dynamics interacting with oscillations of adsorbate coverage on microscale. The surface phase distribution oscillates together with adsorbate coverage, and, in addition, undergoes a slow coarsening process due to the curvature dependence of the drift velocity of interphase boundaries. The coarsening is studied both statistically, assuming a circular shape of islands of the minority phase, and through detailed Lagrangian modeling of boundary dynamics. Direct simulation of boundary dynamics allows us to take into account processes of surface reconstruction, leading to self-induced surface roughening. As a result, the surface becomes inhomogeneous, and the coarsening process is arrested way before the thermodynamic limit is reached, leaving a chaotic distribution of surface phases. (c) 1999 American Institute of Physics. PMID- 12779802 TI - Pattern selection during electropolishing due to double-layer effects. AB - We extend our earlier study of nanoscale pattern formation during electropolishing [Nanotechnology 7, 360 (1996); Phys. Rev. B 56, 12 608 (1997)]. The patterns are attributed to preferential adsorption of organic molecules on the convex portion of the electrode due to its enhanced electric field. This local enhancement occurs because of the effect of surface curvature on the double layer potential drop. By allowing for transport correction to the double-layer potential drop at thermodynamic equilibrium, we estimate this anodic overpotential to be in the realistic mV range and hence verify the Debye-Huckel approximation used in our model. This small anodic overpotential suggests that pattern formation is a generic electropolishing phenomenon whose only requirement is that the polarizability of the organic additive relative to water must lie within a range specified by our theory. We verify this prediction experimentally with a variety of electrolyte solutions. The voltage ranges for specific hexagonal and ridge patterns are well correlated by our model with only a single parameter. (c) 1999 American Institute of Physics. PMID- 12779803 TI - Using weighted global control for stabilizing patterned states. AB - A general method to control a desired patterned state in reaction-diffusion processes is presented. Weighted global control is aimed to keep weighted spatially averaged properties of state variable at preset values. It is shown that weighted global control creates a stable direction in the global space of system states and affects system dynamics globally. We apply it for a specific two-component reaction-diffusion system and show that the desired pattern is attainable for a wide range of the control parameters. (c) 1999 American Institute of Physics. PMID- 12779804 TI - On the dynamics of nonlinear systems with input constraints. AB - In this work we deal with the dynamical analysis of nonlinear systems with input constraints. A characterization of the domain of attraction of the region of controllability of an equilibrium point under bounded control is provided and the concept of regions of invariance within such domains of attraction is introduced and characterized. The concepts and results are illustrated through case studies on chemical reactor models. (c) 1999 American Institute of Physics. PMID- 12779805 TI - Reaction network reduction for distributed systems by model training in lumped reactors: Application to bifurcations in combustion. AB - A new methodology is presented to derive reduced reaction mechanisms for distributed reacting flows by model training in a lumped parameter system (a continuous-stirred tank reactor). The method identifies the relevant transport time scales in the reaction zone of a distributed system along with the local composition vector, over a range of operation conditions. A training box in the parameter space of pressure-transport time scale-composition is then identified. Sensitivity and principal component analyses are subsequently performed at bifurcation points in a lumped parameter system at representative conditions of the training box. The most inclusive chemistry derived in the lumped system captures the proper transport-chemistry coupling and is suitable for the distributed reactor. Application to ignition of hydrogen/air and methane/air mixtures is presented and validated for premixed and diffusion flames in a stagnation flow geometry. (c) 1999 American Institute of Physics. PMID- 12779806 TI - Identification of low order manifolds: Validating the algorithm of Maas and Pope. AB - The algorithm of Maas and Pope (1992) is presented as a method for identification of invariant reduced-order manifolds for stable systems which exhibit dynamics with a time-scale separation. While this method has been published previously in the literature, theoretical justification for the algorithm was not presented in the original work. Here, it will be shown rigorously that the algorithm correctly identifies the slow manifold. Before the theoretical results are presented, a brief background on the behavior of singularly perturbed systems is presented. The algorithm of Maas and Pope (1992) is then introduced. This method will be applied to two different examples, a distillation column and a two-phase chemical reactor. For each of these examples, the resulting reduced-order description will be compared to other standard methods of producing reduced-order models. In addition, some preliminary thoughts on how this method can be used to form reduced-order models are presented. (c) 1999 American Institute of Physics. PMID- 12779807 TI - A tutorial on the Rayleigh-Marangoni-Benard problem with multiple layers and side wall effects. AB - A brief review in the form of a tutorial is presented on convective instabilities that arise from thermocapillary and buoyancy effects. This tutorial primarily focuses on the effect of multiple layers and side walls on the nature of the convective flows and associated patterns. A comprehensive explanation of the physics of this type of convection is followed by a discussion of the mathematical features of bifurcation associated with the problem and some of the recent experimental studies. (c) 1999 American Institute of Physics. PMID- 12779808 TI - Spreading of a surfactant monolayer on a thin liquid film: Onset and evolution of digitated structures. AB - We describe the response of an insoluble surfactant monolayer spreading on the surface of a thin liquid film to small disturbances in the film thickness and surfactant concentration. The surface shear stress, which derives from variations in surfactant concentration at the air-liquid interface, rapidly drives liquid and surfactant from the source toward the distal region of higher surface tension. A previous linear stability analysis of a quasi-steady state solution describing the spreading of a finite strip of surfactant on a thin Newtonian film has predicted only stable modes. [Dynamics in Small Confining Systems III, Materials Research Society Symposium Proceedings, edited by J. M. Drake, J. Klafter, and E. R. Kopelman (Materials Research Society, Boston, 1996), Vol. 464, p. 237; Phys. Fluids A 9, 3645 (1997); O. K. Matar Ph.D. thesis, Princeton University, Princeton, NJ, 1998]. A perturbation analysis of the transient behavior, however, has revealed the possibility of significant amplification of disturbances in the film thickness within an order one shear time after the onset of flow [Phys. Fluids A 10, 1234 (1998); "Transient response of a surfactant monolayer spreading on a thin liquid film: Mechanism for amplification of disturbances," submitted to Phys. Fluids]. In this paper we describe the linearized transient behavior and interpret which physical parameters most strongly affect the disturbance amplification ratio. We show how the disturbances localize behind the moving front and how the inclusion of van der Waals forces further enhances their growth and lifetime. We also present numerical solutions to the fully nonlinear 2D governing equations. As time evolves, the nonlinear system sustains disturbances of longer and longer wavelength, consistent with the quasi-steady state and transient linearized descriptions. In addition, for the parameter set investigated, disturbances consisting of several harmonics of a fundamental wavenumber do not couple significantly. The system eventually singles out the smallest wavenumber disturbance in the chosen set. The summary of results to date seems to suggest that the fingering process may be a transient response which nonetheless has a dramatic influence on the spreading process since the digitated structures redirect the flux of liquid and surfactant to produce nonuniform surface coverage. (c) 1999 American Institute of Physics. PMID- 12779809 TI - The sharkskin instability of polymer melt flows. AB - Flows of polymeric liquids undergo instabilities whose origins are quite different from those of Newtonian flows, due to their elastic character and the complexity of the fluid/solid boundary condition. This article reviews recent studies of one such instability, the sharkskin phenomenon observed during extrusion of many linear polymers. Key experimental observations are summarized; one important fact that has become clear is the importance of the interaction between the molten polymer and the solid walls of the flow channel, especially near the contact line at the exit of the channel. Recent developments in understanding the relationship between wall slip and disentanglement of wall adsorbed polymers from the bulk flow are briefly described, and putative heuristic mechanisms relating the instability to slip and contact line motion are presented. Finally, we review mathematical analyses of the stability of viscoelastic shear flows with slip boundary conditions. Some recent analyses yield instability predictions that are consistent with experiments, but further work is required to discriminate between the various mechanisms that have been proposed. (c) 1999 American Institute of Physics. PMID- 12779810 TI - Contacting and forming singularities: Distinguishing examples. AB - A thin film bridge breaks in a way that starts at one equilibrium state and ends at another equilibrium state. The dynamical trajectory that carries it from connected to disconnected provides rare evidence regarding the singularity of passage through topological change. This nonequilibrium trajectory, called a "forming" flow, is discussed in an attempt to frame it within the larger class of singularities for which bounding surfaces do not remain material surfaces. As a contrast, the weaker "contacting" singularity is illustrated by a stagnation flow where material points reach the stagnation point in finite time. A classification scheme based on pathology of the nonunique Lagrangian motions is suggested. New results for the disconnection example include healing of surgery in post disconnection simulations, different dynamical scalings of the just-disconnected components and a comparison of post-disconnection simulation to experiment. (c) 1999 American Institute of Physics. PMID- 12779811 TI - Residence-time distributions for chaotic flows in pipes. AB - In this paper we derive two rigorous properties of residence-time distributions for flows in pipes and mixers motivated by computational results of Khakhar et al. [Chem. Eng. Sci. 42, 2909 (1987)], using some concepts from ergodic theory. First, a curious similarity between the isoresidence-time plots and Poincare maps of the flow observed in Khakhar et al. is resolved. It is shown that in long pipes and mixers, Poincare maps can serve as a useful guide in the analysis of isoresidence-time plots, but the two are not equivalent. In particular, for long devices isoresidence-time sets are composed of orbits of the Poincare map, but each isoresidence-time set can be comprised of many orbits. Second, we explain the origin of multimodal residence-time distributions for nondiffusive motion of particles in pipes and mixers. It is shown that chaotic regions in the Poincare map contribute peaks to the appropriately defined and rescaled axial distribution functions. (c) 1999 American Institute of Physics. PMID- 12779812 TI - Design criteria of a chemical reactor based on a chaotic flow. AB - We consider the design criteria of a chemical mixing device based on a chaotic flow, with an emphasis on the steady-state devices. The merit of a reactor, defined as the Q-factor, is related to the physical dimension of the device and the molecular diffusivity of the reactants through the local Lyapunov exponents of the flow. The local Lyapunov exponent can be calculated for any given flow field and it can also be measured in experimental situations. Easy-to-compute formulae are provided to estimate the Q-factor given either the exact spatial dependence of the local Lyapunov exponent or its probability distribution function. The requirements for optimization are made precise in the context of local Lyapunov exponents. (c) 1999 American Institute of Physics. PMID- 12779813 TI - Chaotic mixing of granular materials in two-dimensional tumbling mixers. AB - We consider the mixing of similar, cohesionless granular materials in quasi-two dimensional rotating containers by means of theory and experiment. A mathematical model is presented for the flow in containers of arbitrary shape but which are symmetric with respect to rotation by 180 degrees and half-filled with solids. The flow comprises a thin cascading layer at the flat free surface, and a fixed bed which rotates as a solid body. The layer thickness and length change slowly with mixer rotation, but the layer geometry remains similar at all orientations. Flow visualization experiments using glass beads in an elliptical mixer show good agreement with model predictions. Studies of mixing are presented for circular, elliptical, and square containers. The flow in circular containers is steady, and computations involving advection alone (no particle diffusion generated by interparticle collisions) show poor mixing. In contrast, the flow in elliptical and square mixers is time periodic and results in chaotic advection and rapid mixing. Computational evidence for chaos in noncircular mixers is presented in terms of Poincare sections and blob deformation. Poincare sections show regions of regular and chaotic motion, and blobs deform into homoclinic tendrils with an exponential growth of the perimeter length with time. In contrast, in circular mixers, the motion is regular everywhere and the perimeter length increases linearly with time. Including particle diffusion obliterates the typical chaotic structures formed on mixing; predictions of the mixing model including diffusion are in good qualitative and quantitative (in terms of the intensity of segregation variation with time) agreement with experimental results for mixing of an initially circular blob in elliptical and square mixers. Scaling analysis and computations show that mixing in noncircular mixers is faster than that in circular mixers, and the difference in mixing times increases with mixer size. (c) 1999 American Institute of Physics. PMID- 12779814 TI - Observation of structure in the Lorenz map. AB - Through massive numerical integration of the Lorenz system, we are able to discern structure in its Poincare map. We are also able to estimate its capacity dimension; our result is consistent with previous measurements of the correlation dimension of the Lorenz attractor. (c) 1999 American Institute of Physics. PMID- 12779815 TI - Numerical study of reverse period doubling route from chaos to stability in a two mode intracavity doubled Nd-YAG laser. AB - We have numerically studied the behavior of a two-mode Nd-YAG laser with an intracavity KTP crystal. It is found that when the parameter, which is a measure of the relative orientations of the KTP crystal with respect to the Nd-YAG crystal, is varied continuously, the output intensity fluctuations change from chaotic to stable behavior through a sequence of reverse period doubling bifurcations. The graph of the intensity in the X-polarized mode against that in the Y-polarized mode shows a complex pattern in the chaotic regime. The Lyapunov exponent is calculated for the chaotic and periodic regions. (c) 1999 American Institute of Physics. PMID- 12779816 TI - A method for visualization of invariant sets of dynamical systems based on the ergodic partition. AB - We provide an algorithm for visualization of invariant sets of dynamical systems with a smooth invariant measure. The algorithm is based on a constructive proof of the ergodic partition theorem for automorphisms of compact metric spaces. The ergodic partition of a compact metric space A, under the dynamics of a continuous automorphism T, is shown to be the product of measurable partitions of the space induced by the time averages of a set of functions on A. The numerical algorithm consists of computing the time averages of a chosen set of functions and partitioning the phase space into their level sets. The method is applied to the three-dimensional ABC map for which the dynamics was visualized by other methods in Feingold et al. [J. Stat. Phys. 50, 529 (1988)]. (c) 1999 American Institute of Physics. PMID- 12779817 TI - Hyperchaotic qualities of the ball motion in a ball milling device. AB - Ball collisions in milling devices are governed by complex dynamics ruled by impredictable impulsive forces. In this paper, nonlinear dynamics techniques are employed to analyze the time series describing the trajectory of a milling ball in an empty container obtained from a numerical model. The attractor underlying the system dynamics was reconstructed by the time delay method. In order to characterize the system dynamics the calculation of the spectrum of Lyapunov exponents was performed. Six Lyapunov exponents, divided into two terns with opposite sign, were obtained. The detection of the positive tern demonstrates the occurrence of the hyperchaotic qualities of the ball motion. A fractal Lyapunov dimension, equal to 5.62, was also obtained confirming the strange features of the attractor. (c) 1999 American Institute of Physics. PMID- 12779818 TI - Role of multistability in the transition to chaotic phase synchronization. AB - In this paper we describe the transition to phase synchronization for systems of coupled nonlinear oscillators that individually follow the Feigenbaum route to chaos. A nested structure of phase synchronized regions of different attractor families is observed. With this structure, the transition to nonsynchronous behavior is determined by the loss of stability for the most stable synchronous mode. It is shown that the appearance of hyperchaos and the transition from lag synchronization to phase synchronization are related to the merging of chaotic attractors from different families. Numerical examples using Rossler systems and model maps are given. (c) 1999 American Institute of Physics. PMID- 12779819 TI - The control of high-dimensional chaos in time-delay systems to an arbitrary goal dynamics. AB - We present the control of high-dimensional chaos, with possibly a large number of positive Lyapunov exponents, of unknown time-delay systems to an arbitrary goal dynamics. We give an existence-and-uniqueness theorem for the control force. In the case of an unknown system, a formula to compute a model-based control force is derived. We give an example by demonstrating the control of the Mackey-Glass system toward a fixed point and a Rossler dynamics. (c) 1999 American Institute of Physics. PMID- 12779820 TI - Chirality dependent component of vortex advection in excitable media. AB - An advective field induces drift of a vortex in excitable media. The component of the drift velocity C( perpendicular ) perpendicular to the field is known to change its sign with the chirality of the vortex. In an experiment with vortices in an electric field in a chemical excitable medium, we have found unexpectedly that C( perpendicular ) changes its sign also independently of chirality with changing composition of the medium. We did not succeed to explain this phenomenon by using existing mathematical models of chemical excitable media. The experiment described calls for more realistic models.(c) 1999 American Institute of Physics. PMID- 12779821 TI - Dynamics of a nonlinear parametrically excited partial differential equation. AB - We investigate a parametrically excited nonlinear Mathieu equation with damping and limited spatial dependence, using both perturbation theory and numerical integration. The perturbation results predict that, for parameters which lie near the 2:1 resonance tongue of instability corresponding to a single mode of shape cos nx, the resonant mode achieves a stable periodic motion, while all the other modes are predicted to decay to zero. By numerically integrating the p.d.e. as well as a 3-mode o.d.e. truncation, the predictions of perturbation theory are shown to represent an oversimplified picture of the dynamics. In particular it is shown that steady states exist which involve many modes. The dependence of steady state behavior on parameter values and initial conditions is investigated numerically. (c) 1999 American Institute of Physics. PMID- 12779822 TI - Overview: Nonlinear dynamics related to polymeric systems. PMID- 12779823 TI - Beating polymer gels coupled with a nonlinear chemical reaction. AB - We report on a beating polymer gel that exhibits periodical volume changes (swelling and deswelling) in a closed solution without external stimuli, like autonomous heartbeat. The mechanical oscillation is driven by the chemical energy of the oscillatory Belousov-Zhabotinsky (BZ) reaction. The gel is a copolymer gel of N-isopropylacrylamide (NIPAAm) in which ruthenium tris(2,2(')-bipyridine) [Ru(bpy)(3)], known as a catalyst of the BZ reaction, is covalently bonded to the polymer chain. The poly[NIPAAm-co-Ru(bpy)(3)] gel provides an open system where the BZ reaction proceeds, when immersed in an aqueous solution containing the reactants of the BZ reaction (with the exception of a catalyst). The chemical oscillation in the BZ reaction generates the periodical changes of the charge of Ru(bpy)(3) in the gel network between reduced [Ru(II)] and oxidized [Ru(III)] states. The gel swells at the oxidized state because the hydrophilicity of the polymer chains increases, while at the reduced state the gel deswells. Thus, the chemical energy is transduced into the mechanical energy to drive the polymer gel oscillation with a period of about 5 min, depending on the composition of the surrounding solution. The oscillation mode of the gel depends on its size scaled by the wavelength of the BZ pattern. Sufficiently small bead-like gels demonstrate isotropic beating. A large rectangular gel shows mechanical oscillation with a peristaltic motion coupled with the propagating chemical waves. The dynamic behavior of the chemical and mechanical oscillations have been analyzed with a model simulation. (c) 1999 American Institute of Physics. PMID- 12779824 TI - Autonomous gel/enzyme oscillator fueled by glucose: Preliminary evidence for oscillations. AB - A novel prototype gel oscillator that functions by dissipating the chemical energy of glucose by an enzyme-mediated reaction is proposed. The product of the reaction modulates the degree of swelling and hence the permeability of a poly(N isopropylacrylamide-co-methacrylic acid) gel membrane which in turn regulates the flow of substrate to the enzyme. No external energy is required aside from the chemical energy of glucose present externally at constant concentration. A negative chemomechanical feedback loop is established which, coupled with hysteresis in the membrane permeability characteristics, produces pulsing oscillations. In this study, we introduce a simple model which provides guidelines for experimental design, and report preliminary experimental evidence for oscillation. Application of this prototype system to the episodic delivery of drugs and hormones is envisaged. (c) 1999 American Institute of Physics. PMID- 12779825 TI - The guest-induced oscillation of a monolayer composed of polypeptide containing beta-cyclodextrin at the terminal. AB - We prepared a rod-like amphiphile with a molecular recognition end group, alpha helical and hydrophobic poly(gamma-methyl L-glutamate) (PMG) containing hydrophilic beta-cyclodextrin (CyD) as an active end group (PMG-CyD), and formed its monolayer at the n-hexane/water interface. The interfacial pressure (pi)-area (A) isotherms of the monolayer showed that alpha-helix rod of PMG-CyD could be vertically oriented at the oil/water interface, facing the hydrophilic terminal CyD group to the water phase, by increasing the interfacial concentration of the polypeptide. Under the condition 2-p-toludinyl-naphthalene-6-sulfonate (TNS), an intimate guest molecule for the CyD in water was introduced into the water phase beneath the monolayer. Within a minute the monolayer began to oscillate which could be monitored by the rhythmic response of the interfacial pressure of the monolayer. The oscillation continued over ten minutes and then terminated. The mode of the oscillation was found to change with time, i.e., the initial stage showing a periodic sharp reduction in the interfacial pressure (period I), the second stage having sharp increase in the pi value (period II), and the last stage of irregular oscillations (period III). The Fourier analysis of each period also supported the three stages during the oscillatory process. It was also found that when the alpha-helix rod of PMG-CyD lay down in the monolayer, the guest TNS did not induce any changes in the interfacial tension. This nonlinear rhythmic interfacial phenomenon was explained in terms of the periodic movement of the PMG CyD monolayer resulting from the binding and releasing of the guest TNS across the oil/water interface. (c) 1999 American Institute of Physics. PMID- 12779826 TI - Nonlinear effects of electrolyte diodes and transistors in a polymer gel medium. AB - The polarization curve of an acid-base interface in a hydrogel medium has a diode characteristic. Two of each such electrolyte diodes can be combined to give an electrolyte transistor. When a salt is added to the alkaline or to the acidic part of a reverse biased electrolyte diode, the current response is highly nonlinear. If the salt is added to the acidic side, even bistability can be observed. This bistability can generate complex oscillations in a base-acid-base electrolyte transistor. These nonlinear effects are studied experimentally and theoretically. While the nonlinear salt effect can be explained with the Nernst Planck equations, to understand the bistable behavior further investigations are necessary. (c) 1999 American Institute of Physics. PMID- 12779827 TI - Modes selection in polymer mixtures undergoing phase separation by photochemical reactions. AB - Phase separation kinetics and morphology of binary polymer mixtures (A/B) in the presence of photochemical reactions were investigated by using phase-contrast optical microscopy combined with digital image analysis. The polymers were chemically designed in such a way that two types of chemical reactions, intermolecular photodimerization and intramolecular photoisomerization, of polymer segments can be induced and controled by irradiation with ultraviolet light. Unlike the conventional case, the phase separation in the presence of these reactions is spontaneously frozen due to the suppression of the long wavelength instabilities, resulting in stationary spatial structures with intrinsic periodicities. These characteristic length scales are determined by the competition between the two antagonistic interactions: phase separation as a relatively short-range activation and the photochemical reaction as a long-range inhibition. Furthermore, it was found that the spatial symmetry breaking of concentration fluctuations can emerge from the elastic stress associated with the nonhomogeneous kinetics of the reactions. Experimental data obtained with three types of reactions: A-A only cross-link, A-A and B-B simultaneous cross-links and the reversible A<-->B photoisomerization are described. These results do not only indicate that combination of chemical reactions and phase separation could provide a novel method to control the morphology of multiphase polymer materials, but also suggest that photoreactive polymers can be used as a chemical system to study the mode-selection process in polymers far from thermodynamic equilibrium. (c) 1999 American Institute of Physics. PMID- 12779828 TI - Formation of ordered mesoscopic polymer arrays by dewetting. AB - It could be shown that by a simple casting process from solution two dimensionally ordered arrays of mesoscopic (i.e., in the range of submicrometer to micrometer) polymer aggregates on solid substrates can be formed. Patterns were investigated by optical microscopy and atomic force microscopy. The pattern formation was observed in situ by optical and fluorescence microscopy and it was found that a "fingering instability" at the three-phase-line of a solution droplet is the crucial process for pattern formation. (c) 1999 American Institute of Physics. PMID- 12779829 TI - Period-doubling behavior in frontal polymerization of multifunctional acrylates. AB - Front dynamics in the frontal polymerization of two multifunctional acrylate monomers, 1,6-hexanediol diacrylate (HDDA) and trimethylolpropane ethoxylate triacrylate (TMPTA), with Lupersol 231 [1,1-di(t-butylperoxy)-3,3,5 trimethylcyclohexane] as the initiator, are studied. In most frontal polymerization systems, the dynamics are associated with a planar front propagating through the sample. However, in some cases, front behavior can be altered: the front becomes nonplanar characterized by complex patterns like spin modes and pulsations. To determine how these periodic and aperiodic modes arise, reactant solutions consisting of HDDA diluted with diethyl phthalate (DEP) and TMPTA diluted with dimethyl sulfoxide (DMSO) were used in the study. In the study we reveal frontal behavior characteristic of period-doubling behavior, a doubling of spin heads that degenerate into an apparently chaotic mode. Also, a pulsating symmetric mode has been observed. These observations have a striking similarity to observations made in studies of self-propagating high-temperature synthesis (SHS) in which the addition of an inert diluent afforded a rich variety of dynamical behavior. The degree of cross-linking has also been found to be a bifurcation parameter. The energy of activation of multifunctional acrylate polymerization is a strong function of the degree of polymerization. By adding a monoacrylate (benzyl acrylate: BzAc), such that the front temperature was invariant, we observed a period-doubling bifurcation sequence through changes in the energy of activation, which has not been previously reported. (c) 1999 American Institute of Physics. PMID- 12779830 TI - Effect of anisotropy on viscous fingering patterns of polymer solutions in linear Hele-Shaw cells. AB - Viscous fingering patterns of hydroxypropyl methyl cellulose (HPMC) solutions were investigated by forcing air in linear geometry isotropic and anisotropic Hele-Shaw cells as functions of HPMC concentration and injection pressure of air. The resulting patterns depended on the HPMC concentration and the cell anisotropy. The characteristic quantities of pattern growth, such as the finger velocity and the finger width, were evaluated. The finger velocity in the anisotropic linear cell was faster than that in the isotropic one, and the finger velocity and the finger width were well correlated with the pattern morphological changes. (c) 1999 American Institute of Physics. PMID- 12779831 TI - Analytical solutions describing the phase separation driven by a free energy functional containing a long-range interaction term. AB - We are primarily concerned with the variational problem with long-range interaction. This functional represents the Gibbs free energy of the microphase separation of diblock copolymer melts. The critical points of this variational problem can be regarded as the thermodynamic equilibrium state of the phase separation phenomenon. Experimentally it is well-known in the diblock copolymer problem that the final equilibrium state prefers periodic structures such as lamellar, column, spherical, double-diamond geometries and so on. We are interested in the characterization of the periodic structure of the global minimizer of the functional (corresponding to the strong segregation limit). In this paper we completely determine the principal part of the asymptotic expansion of the period with respect to epsilon (interfacial thickness), namely, we estimate the higher order error term of the period with respect to epsilon in a mathematically rigorous way in one space dimension. Moreover, we decide clearly the dependency of the constant of proportion upon the ratio of the length of two homopolymers and upon the quench depth. In the last section, we study the time evolution of the system. We first study the linear stability of spatially homogeneous steady state and derive the most unstable wavelength, if it is unstable. This is related to spinodal decomposition. Then, we numerically investigate the time evolution equation (the gradient flow of the free energy), and see that the free energy has many local minimizers and the system have some kind of sensitivity about initial data. (c) 1999 American Institute of Physics. PMID- 12779832 TI - The autowave modes of solid phase polymerization of metal-containing monomers in two- and three-dimensional fiberglass-filled matrices. AB - The phenomenon of autowave (frontal) solid phase polymerization of metal containing monomers based on metal-acrylamide complexes is considered. The comparison of the features of autowave processes realized in both the single component matrices of the monomer and the matrices filled by the fiberglass materials is performed. The unstable regimes of the polymerization wave as well as the conditions for the stabilization of the flat front in the filled matrices are described. The peculiarities of the frontal regimes in the three- and two dimensional media are studied. Some possibilities for using of autowave polymerization in the fabrication of the polymer-fiberglass composites and composition prepregs are discussed. (c) 1999 American Institute of Physics. PMID- 12779833 TI - On the local stability of limit cycles. AB - Orbital stability of limit cycles is the result of the competing local tendencies of perturbations from the cycle to decay (during phases of local stability) and to grow (during phases of local instability), averaged over a cycle. We examine this coexistence of attractive and repulsive phases on limit cycles, including the local rates of expansion and contraction of phase space volumes. This is done in a frame of reference that moves along the orbit, to partially decouple motions tangential and perpendicular to the cycle. Dynamical systems used for illustration are the generalized Bonhoeffer-van-der-Pol and Rossler models, both far from and near to different types of bifurcations. Finally, it is shown that the nonuniformity of local stability in phase space affects the response of limit cycle oscillators to perturbations and gives rise to their phase-dependent response. (c) 1999 American Institute of Physics. PMID- 12779834 TI - Theory and examples of the inverse Frobenius-Perron problem for complete chaotic maps. AB - The general solution of the inverse Frobenius-Perron problem considering the construction of a fully chaotic dynamical system with given invariant density is obtained for the class of one-dimensional unimodal complete chaotic maps. Some interesting connections between this general solution and the special approach via conjugation transformations are illuminated. The developed method is applied to obtain a class of maps having as invariant density the two-parametric beta probability density function. Varying the parameters of the density a rich variety of dynamics is observed. Observables like autocorrelation functions, power spectra, and Liapunov exponents are calculated for representatives of this family of maps and some theoretical predictions concerning the decay of correlations are tested. (c) 1999 American Institute of Physics. PMID- 12779835 TI - Basin boundaries and focal points in a map coming from Bairstow's method. AB - This paper is devoted to the study of the global dynamical properties of a two dimensional noninvertible map, with a denominator which can vanish, obtained by applying Bairstow's method to a cubic polynomial. It is shown that the complicated structure of the basins of attraction of the fixed points is due to the existence of singularities such as sets of nondefinition, focal points, and prefocal curves, which are specific to maps with a vanishing denominator, and have been recently introduced in the literature. Some global bifurcations that change the qualitative structure of the basin boundaries, are explained in terms of contacts among these singularities. The techniques used in this paper put in evidence some new dynamic behaviors and bifurcations, which are peculiar of maps with denominator; hence they can be applied to the analysis of other classes of maps coming from iterative algorithms (based on Newton's method, or others). (c) 1999 American Institute of Physics. PMID- 12779836 TI - Diffusion and scaling in escapes from two-degrees-of-freedom Hamiltonian systems. AB - This paper summarizes an investigation of the statistical properties of orbits escaping from three different two-degrees-of-freedom Hamiltonian systems which exhibit global stochasticity. Each time-independent H=H(0)+ varepsilon H('), with H(0) an integrable Hamiltonian and varepsilon H(') a nonintegrable correction, not necessarily small. Despite possessing very different symmetries, ensembles of orbits in all three potentials exhibit similar behavior. For varepsilon below a critical varepsilon (0), escapes are impossible energetically. For somewhat higher values, escape is allowed energetically but still many orbits never escape. The escape probability P computed for an arbitrary orbit ensemble decays toward zero exponentially. At or near a critical value varepsilon (1)> varepsilon (0) there is a rather abrupt qualitative change in behavior. Above varepsilon (1), P typically exhibits (1) an initial rapid evolution toward a nonzero P(0)( varepsilon ), the value of which is independent of the detailed choice of initial conditions, followed by (2) a much slower subsequent decay toward zero which, in at least one case, is well fit by a power law P(t) proportional, variant t( &mgr;), with &mgr; approximately 0.35-0.40. In all three cases, P(0) and the time T required to converge toward P(0) scale as powers of varepsilon - varepsilon (1), i.e., P(0) proportional, variant ( varepsilon - varepsilon (1))(alpha) and T proportional, variant ( varepsilon - varepsilon (1))(beta), and T also scales in the linear size r of the region sampled for initial conditions, i.e., T proportional, variant r(-delta). To within statistical uncertainties, the best fit values of the critical exponents alpha, beta, and delta appear to be the same for all three potentials, namely alpha approximately 0.5, beta approximately 0.4, and delta approximately 0.1, and satisfy alpha-beta-delta approximately 0. The transitional behavior observed near varepsilon (1) is attributed to the breakdown of some especially significant KAM tori or cantori. The power law behavior at late times is interpreted as reflecting intrinsic diffusion of chaotic orbits through cantori surrounding islands of regular orbits. (c) 1999 American Institute of Physics. PMID- 12779837 TI - Numerical study of chaos based on a shell model. AB - A shell model is introduced to study a turbulence driven by the thermal instability (Rayleigh-Benard convection). This model equation describes cascade and chaos in the strong turbulence with high Rayleigh number. The chaos is numerically studied based on this model. The characteristics of the turbulence are analyzed and compared with those of the Gledzer-Ohkitani-Yamada (GOY) model. Quantities such as a mean value of total fluctuation energy, it's standard deviation, time averaged wave spectrum, probability distribution function, frequency spectrum, the maximum instantaneous Lyapunov exponent, distribution of instantaneous Lyapunov exponents, are evaluated. The dependences of these quantities on the error of numerical integration are also examined. There is not a clear correlation between the numerical accuracy and the accuracy of these quantities, since the interaction between a truncation error and an intrinsic nonlinearity of the system exists. A finding is that the maximum Lyapunov exponent is insensitive to a truncation error. (c) 1999 American Institute of Physics. PMID- 12779838 TI - Bifurcations and selection of equilibria in a simple cosymmetric model of filtrational convection. AB - A three-dimensional set of ordinary differential equations that constitutes a simple abstract model of Darcy convection is investigated. The model reproduces a number of effects that are typical for dynamic systems with nontrivial cosymmetry. Nontrivial cosymmetry can give rise to a continuous family of equilibria where, in this case, the equilibrium stability spectrum varies along the family. The family of equilibria and its stability are examined analytically, and special bifurcations that occur in the system are investigated. It is shown that discrete and continual symmetries, called "flash symmetries," can be present in the system for certain parameter values. Computer experiments on the selection of equilibria in the symmetric and cosymmetric cases have been carried out. They showed that, for initial points that are far enough from a cycle of equilibria, the neighborhood of a single equilibrium is established in the case of cosymmetry, but all the equilibria are equivalent in the case of symmetry. The authors hope that these results, as well as the formulation of the problems and the approach to their solution, will serve as a sample in the investigation of more complex systems in mathematical physics. (c) 1999 American Institute of Physics. PMID- 12779839 TI - Practical implementation of nonlinear time series methods: The TISEAN package. AB - We describe the implementation of methods of nonlinear time series analysis which are based on the paradigm of deterministic chaos. A variety of algorithms for data representation, prediction, noise reduction, dimension and Lyapunov estimation, and nonlinearity testing are discussed with particular emphasis on issues of implementation and choice of parameters. Computer programs that implement the resulting strategies are publicly available as the TISEAN software package. The use of each algorithm will be illustrated with a typical application. As to the theoretical background, we will essentially give pointers to the literature. (c) 1999 American Institute of Physics. PMID- 12779840 TI - Estimating the amplitude of measurement noise present in chaotic time series. AB - We propose a method for estimating the amplitude of measurement noise present in chaotic time series. This method is based on the evaluation of initial errors for a given time series and for a new one synthesized by adding an adequate amount of noise to the given one. The method is valid over a much wider range of noise levels than the previous methods are because it is not based on the detail of dynamical structure which generates the data. In addition, it is possible to check if the method is valid for the given data prior to its application. To confirm the effectiveness of the method we show the results of numerical experiments and apply the method to chaotic data obtained from an electrochemical experiment. (c) 1999 American Institute of Physics. PMID- 12779841 TI - Using multiple attractor chaotic systems for communication. AB - In recent work with symmetric chaotic systems, we synchronized two such systems with one-way driving. The drive system had two possible attractors, but the response system always synchronized with the drive system. In this work, we show how we may combine two attractor chaotic systems with a multiplexing technique first developed by Tsimring and Suschick to make a simple communications system. We note that our response system is never synchronized to our drive system (not even in a generalized sense), but we are still able to transmit information. We characterize the performance of the communications system when noise is added to the transmitted signal. PMID- 12779842 TI - Scale and space localization in the Kuramoto-Sivashinsky equation. AB - We describe a wavelet-based approach to the investigation of spatiotemporally complex dynamics, and show through extensive numerical studies that the dynamics of the Kuramoto-Sivashinsky equation in the spatiotemporally chaotic regime may be understood in terms of localized dynamics in both space and scale (wave number). A projection onto a spline wavelet basis enables good separation of scales, each with characteristic dynamics. At the large scales, one observes essentially slow Gaussian dynamics; at the active scales, structured "events" reminiscent of traveling waves and heteroclinic cycles appear to dominate; while the strongly damped small scales display intermittent behavior. The separation of scales and their dynamics is invariant as the length of the system increases, providing additional support for the extensivity of the spatiotemporally complex dynamics claimed in earlier works. We show also that the dynamics are spatially localized, discuss various correlation lengths, and demonstrate the existence of a characteristic interaction length for instantaneous influences. Our results motivate and advance the search for localized, low-dimensional models that capture the full behavior of spatially extended chaotic partial differential equations. (c) 1999 American Institute of Physics. PMID- 12779843 TI - Scaling and interleaving of subsystem Lyapunov exponents for spatio-temporal systems. AB - The computation of the entire Lyapunov spectrum for extended dynamical systems is a very time consuming task. If the system is in a chaotic spatio-temporal regime it is possible to approximately reconstruct the Lyapunov spectrum from the spectrum of a subsystem by a suitable rescaling in a very cost effective way. We compute the Lyapunov spectrum for the subsystem by truncating the original Jacobian without modifying the original dynamics and thus taking into account only a portion of the information of the entire system. In doing so we notice that the Lyapunov spectra for consecutive subsystem sizes are interleaved and we discuss the possible ways in which this may arise. We also present a new rescaling method, which gives a significantly better fit to the original Lyapunov spectrum. We evaluate the performance of our rescaling method by comparing it to the conventional rescaling (dividing by the relative subsystem volume) for one- and two-dimensional lattices in spatio-temporal chaotic regimes. Finally, we use the new rescaling to approximate quantities derived from the Lyapunov spectrum (largest Lyapunov exponent, Lyapunov dimension, and Kolmogorov-Sinai entropy), finding better convergence as the subsystem size is increased than with conventional rescaling. (c) 1999 American Institute of Physics. PMID- 12779844 TI - Wave analysis of ray chaos in underwater acoustics. AB - The dispersion of a wave packet in an acoustic medium is considered in the paraxial wave approximation, where the effective potential, due to variation of the speed of propagation, varies both with depth and propagation distance. The analysis of the resulting parabolic equation, similar to the Schrodinger equation, clearly demonstrates the role of ray chaos in enhancing the dispersion of the initial packet. However, wave coherence effects are also seen that suppress the effects of the ray chaos in a manner analogous to the effects of quantum chaos. (c) 1999 American Institute of Physics. PMID- 12779845 TI - Chaos in spacecraft attitude motion in Earth's magnetic field. AB - The rotational motion of a satellite with a magnetic stabilization system is discussed. The motion is described by a nonautonomous differential equation, with the magnetic moment of the satellite as a parameter. The global phase portrait of the problem is investigated in a wide range of magnetic-parameter values, using a numerical realization of the method of Poincare point maps. New periodic solutions of the problem are found, and an analysis is carried out of the evolution of the phase portrait and the bifurcation of periodic solutions with varying magnetic-parameter values. The values of the magnetic parameter that must be avoided in the design of the satellite magnetic stabilization system are discussed. (c) 1999 American Institute of Physics. PMID- 12779846 TI - Noise and O(1) amplitude effects on heteroclinic cycles. AB - The dynamics of structurally stable heteroclinic cycles connecting fixed points with one-dimensional unstable manifolds under the influence of noise is analyzed. Fokker-Planck equations for the evolution of the probability distribution of trajectories near heteroclinic cycles are solved. The influence of the magnitude of the stable and unstable eigenvalues at the fixed points and of the amplitude of the added noise on the location and shape of the probability distribution is determined. As a consequence, the jumping of solution trajectories in and out of invariant subspaces of the deterministic system can be explained. (c) 1999 American Institute of Physics. PMID- 12779847 TI - Introduction to the focus issue on granular materials. AB - In a review paper [H. M. Jaeger, S. R. Nagel, and R. P. Behringer, "Granular solids, liquids and gases," Rev. Mod. Phys. 68, 1259-1273 (1996)] a few years ago, we wrote about granular material as a distinctive form of matter that exhibits behavior rather different from that of ordinary solids, liquids, or gases. We traced this distinction to three characteristic properties. First, the individual particles making up a granular material are typically large so that thermal energy is irrelevant compared to gravitational energy. Consequently, concepts from equilibrium statistical mechanics are often not applicable. Second, the interactions between particles are frictional and can be mobilized to different degrees depending on the preparation history, giving rise to memory effects, i.e., a static pile will remember how it was formed. Third, when particles collide they do so inelastically so that a "gas" of particles will slow down and come to rest in clumps. In the intervening years, the research on granular matter has progressed rapidly and this may be a good time to ask what we have learned since that article was written. In this spirit, the present special issue of the journal Chaos assembles a spectrum of papers discussing recent developments in the field. (c) 1999 American Institute of Physics. PMID- 12779848 TI - Jamming and static stress transmission in granular materials. AB - We have recently developed some simple continuum models of static granular media which display "fragile" behavior: They predict that the medium is unable to support certain types of infinitesimal load (which we call "incompatible" loads) without plastic rearrangement. We argue that a fragile description may be appropriate when the mechanical integrity of the medium arises adaptively, in response to a load, through an internal jamming process. We hypothesize that a network of force chains (or "granular skeleton") evolves until it can just support the applied load, at which point it comes to rest; it then remains intact so long as no incompatible load is applied. Our fragile models exhibits unusual mechanical responses involving hyperbolic equations for stress propagation along fixed characteristics through the material. These characteristics represent force chains; their arrangement expressly depends on the construction history. Thus, for example, we predict a large difference in the stress pattern beneath two conical piles of sand, one poured from a point source and one created by sieving. (c) 1999 American Institute of Physics. PMID- 12779849 TI - Modeling of stress distribution in granular piles: Comparison with centrifuge experiments. AB - The classical method to compute stress and strain distributions in granular materials is recalled using continuum mechanics approach, and different rheological laws described. It is recalled that granular materials exhibit highly nonlinear response such as nonlinear elasticity, dilatancy and plastic flow. Finite element technique is used to predict the stress field distribution below a conic and a triangular pile. The dependence of the stress distribution on the rheological law, the bottom boundary condition and the building process (horizontal or inclined strata) is demonstrated. These results are compared to experimental data obtained in centrifuge. (c) 1999 American Institute of Physics. PMID- 12779850 TI - Contact forces in a granular packing. AB - We present the results of a systematic numerical investigation of force distributions in granular packings. We find that all the main features of force transmission previously established for two-dimensional systems of hard particles hold in three-dimensional systems and for soft particles, too. In particular, the probability distribution of normal forces falls off exponentially for forces above the mean force. For forces below the mean, this distribution is either a decreasing power law when the system is far from static equilibrium, or nearly uniform at static equilibrium, in agreement with recent experiments. Moreover, we show that the forces below the mean do not contribute to the shear stress. The subnetwork of the contacts carrying a force below the mean thus plays a role similar to a fluid surrounding the solid backbone composed of the contacts carrying a force above the mean. We address the issue of the computation of contact forces in a packing at static equilibrium. We introduce a model with no local simplifying force rules, that allows for an exact computation of contact forces for given granular texture and boundary conditions. (c) 1999 American Institute of Physics. PMID- 12779851 TI - Compactivity and transmission of stress in granular materials. AB - We outline a statistical-mechanical theory of granular materials. Stress propagation and force fluctuations in static granular media are still poorly understood. We develop the statistical-mechanical theory that delivers the fundamental equations of stress equilibrium. The formalism is based on the assumptions that grains are rigid, cohesionless, and that friction is perfect. Since grains are assumed perfectly rigid, no strain or displacement field can enter the equations for static equilibrium of the stress field. The complete system of equations for the stress tensor is derived from the equations of intergranular force and torque balance, given the geometric specification of the material. These new constitutive equations are indeed fundamental and are based on relations between various components of the stress tensor within the material, and depend on the topology of the granular packing. The problem of incorporating into the formalism the "no tensile forces" constraint is considered. The compactivity concept is reviewed. We discuss the relation between the concept of compactivity and the problem of stress transmission. (c) 1999 American Institute of Physics. PMID- 12779852 TI - Fluctuations in granular media. AB - Dense slowly evolving or static granular materials exhibit strong force fluctuations even though the spatial disorder of the grains is relatively weak. Typically, forces are carried preferentially along a network of "force chains." These consist of linearly aligned grains with larger-than-average force. A growing body of work has explored the nature of these fluctuations. We first briefly review recent work concerning stress fluctuations. We then focus on a series of experiments in both two- and three-dimension [(2D) and (3D)] to characterize force fluctuations in slowly sheared systems. Both sets of experiments show strong temporal fluctuations in the local stress/force; the length scales of these fluctuations extend up to 10(2) grains. In 2D, we use photoelastic disks that permit visualization of the internal force structure. From this we can make comparisons to recent models and calculations that predict the distributions of forces. Typically, these models indicate that the distributions should fall off exponentially at large force. We find in the experiments that the force distributions change systematically as we change the mean packing fraction, gamma. For gamma's typical of dense packings of nondeformable grains, we see distributions that are consistent with an exponential decrease at large forces. For both lower and higher gamma, the observed force distributions appear to differ from this prediction, with a more Gaussian distribution at larger gamma and perhaps a power law at lower gamma. For high gamma, the distributions differ from this prediction because the grains begin to deform, allowing more grains to carry the applied force, and causing the distributions to have a local maximum at nonzero force. It is less clear why the distributions differ from the models at lower gamma. An exploration in gamma has led to the discovery of an interesting continuous or "critical" transition (the strengthening/softening transition) in which the mean stress is the order parameter, and the mean packing fraction, gamma, must be adjusted to a value gamma(c) to reach the "critical point." We also follow the motion of individual disks and obtain detailed statistical information on the kinematics, including velocities and particle rotations or spin. Distributions for the azimuthal velocity, V(theta), and spin, S, of the particles are nearly rate invariant, which is consistent with conventional wisdom. Near gamma(c), the grain motion becomes intermittent causing the mean velocity of grains to slow down. Also, the length of stress chains grows as gamma-->gamma(c). The 3D experiments show statistical rate invariance for the stress in the sense that when the power spectra and spectral frequencies of the stress time series are appropriately scaled by the shear rate, Omega, all spectra collapse onto a single curve for given particle and sample sizes. The frequency dependence of the spectra can be characterized by two different power laws, P proportional, variant omega(-alpha), in the high and low frequency regimes: alpha approximately 2 at high omega; alpha<2 at low omega. The force distributions computed from the 3D stress time series are at least qualitatively consistent with exponential fall-off at large stresses. (c) 1999 American Institute of Physics. PMID- 12779853 TI - Axial segregation of granular materials. AB - When mixtures of granular materials are rotated, it is often found that they segregate into bands, along the axis of rotation, which are rich in the various components. This effect is discussed experimentally and theoretically, with emphasis on a mechanism based on surface flow. The complimentary phenomenon of radial segregation is reviewed, and a mechanism is proposed. Finally, we consider the long-time behavior of rotating mixtures, particularly their anomolous coarsening. (c) 1999 American Institute of Physics. PMID- 12779854 TI - Measurement of particle motions within tumbling granular flows. AB - Flowing granular materials are complex, industrially important, and scientifically provocative. In this paper we report measurements of granular transport in 3-dimensional tumbling containers. We use magnetic resonance imaging techniques for direct tracking of particles and measure the interior flows of granular materials. One goal is to measure industrial mixer performance over a wide range of conditions. As the mixer geometries are relatively simple, such measurements could serve as incisive tests during development of better granular equations of motion. (c) 1999 American Institute of Physics. PMID- 12779855 TI - Mixing and segregation of granular materials in chute flows. AB - Mixing of granular solids is invariably accompanied by segregation, however, the fundamentals of the process are not well understood. We analyze density and size segregation in a chute flow of cohesionless spherical particles by means of computations and theory based on the transport equations for a mixture of nearly elastic particles. Computations for elastic particles (Monte Carlo simulations), nearly elastic particles, and inelastic, frictional particles (particle dynamics simulations) are carried out. General expressions for the segregation fluxes due to pressure gradients and temperature gradients are derived. Simplified equations are obtained for the limiting cases of low volume fractions (ideal gas limit) and equal sized particles. Theoretical predictions of equilibrium number density profiles are in good agreement with computations for mixtures of equal sized particles with different density for all solids volume fractions, and for mixtures of different sized particles at low volume fractions (nu<0.2), when the particles are elastic or nearly elastic. In the case of inelastic, frictional particles the theory gives reasonable predictions if an appropriate effective granular temperature is assumed. The relative importance of pressure diffusion and temperature diffusion for the cases considered is discussed. (c) 1999 American Institute of Physics. PMID- 12779856 TI - Chaotic granular mixing. AB - Several models for convective mixing of coarse, freely flowing in granular tumblers have been proposed over the past decade. Powders of practical interest, by contrast, are frequently fine and cohesive, and cannot be analyzed with these models. Moreover, even in the freely flowing regime, mixing transverse to the dominant, convective, direction is typically slow and inefficient. In this paper, we examine two chaotic mixing mechanisms, the first of which can be intentionally applied to increase transverse mixing rates severalfold, with new prospects for further improvements in three-dimensional mixing through judicious process design. The second mechanism occurs spontaneously in fine grains, resulting in mixing rates overwhelmingly exceeding what would be possible in freely flowing grains. Finally, we show that the same chaotic mixing mechanisms seen in simple drum mixers are also found to be at work in more complex blender configurations widely used in batch industrial operations. (c) 1999 American Institute of Physics. PMID- 12779857 TI - Segregation induced instabilities of granular fronts. AB - Experimental investigation of granular flows containing particles of several sizes and moving down slopes shows that segregation of coarse-grained, irregularly shaped particles induces a fingering instability at the propagating front. The size-segregation mechanism involves percolation of small particles downward and a corresponding migration of large ones toward the flow surface. Large particles at the flow surface experience velocities that are greater than average so that they migrate forward and begin to collect at the flow front. In the case of dry cohesionless flows, the instability depends upon these large particles at the flow perimeter being more angular and thus more resistant to flow than the smaller rounder ones in the interior. A simple analytical model predicts the fingering instability when friction of the flow front is greater than that of the following flow. The presence of viscous liquid inhibits both size-segregation and the development of the instability. Fluidization of dry flows permits segregation of large particles to flow perimeters, thus increasing permeability and permitting a similar instability that owes its development to the dry frictional perimeter that surrounds a partly fluidized interior. (c) 1999 American Institute of Physics. PMID- 12779858 TI - The rotating bucket of sand: Experiment and theory. AB - The surface shape of a bucket of sand rotating about its cylindrical axis is studied experimentally and theoretically. Focusing on fast time scales on which surface shape is determined by avalanches, we identify three regimes of behavior. At intermediate and high frequencies, the surface shape is always at its critical shape determined by the Coulomb yield condition. The low frequency behavior displays an unexpected subcritical region at the center of the bucket. To understand this central region, we adapt a continuum model of surface flow developed by Bouchaud et al. and Mehta et al. The model indicates that the subcritical region is due to a nonlinear instability mechanism. (c) 1999 American Institute of Physics. PMID- 12779859 TI - Motion of grains down a bumpy surface. AB - We summarize in this article an extensive experimental and theoretical effort carried out to understand the behavior of a single ball when rolling down a bumpy surface. This may appear to be a simple problem but in fact is one that displays a rich variety of different behaviors which allow us to understand better dissipative systems such as granular media. Studies performed previously have shown that the motion of the single ball on the rough surface can be characterized by three different dynamic regimes according to the different values of the two control parameters, the inclination angle theta and the ratio Phi=R/r, where R is the radius of the rolling ball and r the radius of the glass beads which make up the rough surface. The three regimes are a decelerated regime A, a stationary regime B, characterized by a constant average velocity and a jumping regime C. This result was found to be independent of the composition of the rolling ball and the rough surface. It has been demonstrated that regime B is characterized by a viscous-like friction force that appears for specific parameter values. This friction force can be explained by a model whose central ingredient is the geometry of the surface. The trajectory of the ball in regime B can be pictured as a driven random walk motion where the fluctuations of the local velocities are due to collisions of the moving sphere and the surface grains. A detailed analysis of diffusive properties of the motion is discussed. (c) 1999 American Institute of Physics. PMID- 12779860 TI - Hysteretic transition between avalanches and continuous flow in rotated granular systems. AB - Experiments in drums or cylinders partly filled with a granular system and rotated constantly about their horizontally aligned axis of symmetry show a hysteretic transition from discrete avalanches to continuous flow if the rotation rate is adiabatically changed. Herein, we show that this hysteresis can be explained by the impact of global Langevin-type fluctuations in a recently proposed minimal model for surface flow along granular piles. For too large magnitudes of the fluctuations corresponding to almost elastic grains, the hysteresis vanishes. This might explain why molecular dynamical simulations were not yet able to detect the hysteretic transition. (c) 1999 American Institute of Physics. PMID- 12779861 TI - Hydraulic theory for a debris flow supported on a collisional shear layer. AB - We consider a heap of grains driven by gravity down an incline. We assume that the heap is supported at its base on a relatively thin carpet of intensely sheared, highly agitated grains that interact through collisions. We adopt the balance laws, constitutive relations, and boundary conditions of a kinetic theory for dense granular flows and determine the relationship between the shear stress, normal stress, and relative velocity of the boundaries in the shear layer in an analysis of a steady shearing flow between identical bumpy boundaries. This relationship permits us to close the hydraulic equations governing the evolution of the shape of the heap and the velocity distribution at its base. We integrate the resulting equations numerically for typical values of the parameters for glass spheres. (c) 1999 American Institute of Physics. PMID- 12779862 TI - Scales and kinetics of granular flows. AB - When a granular material experiences strong forcing, as may be the case, e.g., for coal or gravel flowing down a chute or snow (or rocks) avalanching down a mountain slope, the individual grains interact by nearly instantaneous collisions, much like in the classical model of a gas. The dissipative nature of the particle collisions renders this analogy incomplete and is the source of a number of phenomena which are peculiar to "granular gases," such as clustering and collapse. In addition, the inelasticity of the collisions is the reason that granular gases, unlike atomic ones, lack temporal and spatial scale separation, a fact manifested by macroscopic mean free paths, scale dependent stresses, "macroscopic measurability" of "microscopic fluctuations" and observability of the effects of the Burnett and super-Burnett "corrections." The latter features may also exist in atomic fluids but they are observable there only under extreme conditions. Clustering, collapse and a kinetic theory for rapid flows of dilute granular systems, including a derivation of boundary conditions, are described alongside the mesoscopic properties of these systems with emphasis on the effects, theoretical conclusions and restrictions imposed by the lack of scale separation. (c) 1999 American Institute of Physics. PMID- 12779863 TI - Cluster-growth in freely cooling granular media. AB - When dissipative particles are left alone, their fluctuation energy decays due to collisional interactions, clusters build up and grow with time until the system size is reached. When the effective dissipation is strong enough, this may lead to the "inelastic collapse," i.e., the divergence of the collision frequency of some particles. The cluster growth is an interesting physical phenomenon, whereas the inelastic collapse is an intrinsic effect of the inelastic hard sphere (IHS) model used to study the cluster growth-involving only a negligible number of particles in the system. Here, we extend the IHS model by introducing an elastic contact energy and the related contact duration t(c). This avoids the inelastic collapse and allows to examine the long-time behavior of the system. For a quantitative description of the cluster growth, we propose a burning-like algorithm in continuous space, that readily identifies all particles that belong to the same cluster. The criterion for this is here chosen to be only the particle distance. With this method we identify three regimes of behavior. First, for short times a homogeneous cooling state (HCS) exists, where a mean-field theory works nicely, and the clusters are tiny and grow very slowly. Second, at a certain time which depends on the system's properties, cluster growth starts and the clusters increase in size and mass until, in the third regime, the system size is reached and most of the particles are collected in one huge cluster. (c) 1999 American Institute of Physics. PMID- 12779864 TI - Velocity statistics in excited granular media. AB - We present an experimental study of velocity statistics for a partial layer of inelastic colliding beads driven by a vertically oscillating boundary. Over a wide range of parameters (accelerations 3-8 times the gravitational acceleration), the probability distribution P(v) deviates measurably from a Gaussian for the two horizontal velocity components. It can be described by P(v) approximately exp(-mid R:v/v(c)mid R:(1.5)), in agreement with a recent theory. The characteristic velocity v(c) is proportional to the peak velocity of the boundary. The granular temperature, defined as the mean square particle velocity, varies with particle density and exhibits a maximum at intermediate densities. On the other hand, for free cooling in the absence of excitation, we find an exponential velocity distribution. Finally, we examine the sharing of energy between particles of different mass. The more massive particles are found to have greater kinetic energy. (c) 1999 American Institute of Physics. PMID- 12779865 TI - Convection in horizontally vibrated granular material. AB - We report observations of convective motion in a container filled with granular material when it is vibrated in the horizontal direction. We find that the roughness of the boundaries and the container dimensions play an important role in determining the shape and number of the convection cells. When the container bottom and lateral walls are rough, the system typically exhibits four counter rotating rolls stacked in two pairs on top of each other; for very low filling height, it is possible to observe a single row of rolls arranged laterally along the bottom of the container. With smooth walls, on the other hand, we find that the system typically forms only a single pair of counter-rotating convection rolls that originate in the two upper corners of the vibrated material; when the filling height is increased to a level that depends on the container width, we observe a transition to the four-roll state. (c) 1999 American Institute of Physics. PMID- 12779866 TI - Islands of accelerator modes and homoclinic tangles. AB - Islands are divided according to their phase space structure-resonant islands and tangle islands are considered. It is proved that in the near-integrable limit these correspond to two distinct sets, hence that in general their definitions are not trivially equivalent. It is demonstrated and proved that accelerator modes of the standard map and of the web map are necessarily of the tangle island category. These islands have an important role in determining transport-indeed it has been demonstrated in various works that stickiness to these accelerator modes may cause anomalous transport even for initial conditions starting in the ergodic component. (c) 1999 American Institute of Physics. PMID- 12779867 TI - From scattering singularities to the partition of a horseshoe. AB - In a chaotic scattering system there are two different approaches to construct a symbolic dynamics. One comes from the branching tree obtained from a scattering function. The other comes from a Markov partition based on the line of primary homoclinic tangencies in the Poincare map taken in the interaction region. In general the two results only coincide for a complete horseshoe. We show how to make a different choice for the partition in the internal Poincare section based on scattering behavior and not on homoclinic tangencies. Then the corresponding symbolic dynamics coincides also for the incomplete case with the one obtained naturally from the scattering functions. The scattering based partition lines of the horseshoe are constructed by an iterative procedure. (c) 1999 American Institute of Physics. PMID- 12779868 TI - Statistics of return times in a self-similar model. AB - In this paper, we propose a model for the hyperbolic part of the phase space of a Hamiltonian system that is located near a hierarchical islands around islands structure. We study the statistics of Poincare recurrences, the type of orbits that are mainly responsible for slowly decaying correlations, and thus the mechanisms generating power law tails. (c) 1999 American Institute of Physics. PMID- 12779869 TI - Spectral properties of dissipative chaotic quantum maps. AB - I examine spectral properties of a dissipative chaotic quantum map with the help of a recently discovered semiclassical trace formula. I show that in the presence of a small amount of dissipation the traces of any finite power of the propagator of the reduced density matrix, and traces of its classical counterpart, the Frobenius-Perron operator, are identical in the limit of variant Planck's over 2pi -->0. Numerically I find that even for finite variant Planck's over 2pi the agreement can be very good. This holds in particular if the classical phase space contains a strange attractor, as long as one stays clear of bifurcations. Traces of the quantum propagator for iterations of the map agree well with the corresponding traces of the Frobenius-Perron operator if the classical dynamics is dominated by a strong point attractor. (c) 1999 American Institute of Physics. PMID- 12779870 TI - Synchronized family dynamics in globally coupled maps. AB - The dynamics of a globally coupled, logistic map lattice is explored over a parameter plane consisting of the coupling strength, varepsilon, and the map parameter, a. By considering simple periodic orbits of relatively small lattices, and then an extensive set of initial-value calculations, the phenomenology of solutions over the parameter plane is broadly classified. The lattice possesses many stable solutions, except for sufficiently large coupling strengths, where the lattice elements always synchronize, and for small map parameter, where only simple fixed points are found. For smaller varepsilon and larger a, there is a portion of the parameter plane in which chaotic, asynchronous lattices are found. Over much of the parameter plane, lattices converge to states in which the maps are partitioned into a number of synchronized families. The dynamics and stability of two-family states (solutions partitioned into two families) are explored in detail. (c) 1999 American Institute of Physics. PMID- 12779871 TI - Modal decomposition of hopping states in cellular flames. AB - We use Karhunen-Loeve (KL) decomposition of video images from an experiment to analyze a spatiotemporal dynamic state, unique to cellular flames, referred to as a "hopping state." Ordered states of cellular flames on a circular burner consist of one or two concentric rings of luminous cells. The hopping states correspond to the motions of individual cells in a ring sequentially executing abrupt changes in their angular position, while the other cells in the ring remain symmetric and at rest. KL decomposition separates the spatial and temporal characteristics of the hopping motion. The underlying symmetries of the experiment allow us to deduce a set of normal form equations that describe the formation of these states. We find that they result from secondary bifurcations connecting two primary branches of traveling waves. The solutions corresponding to hopping states exist as mixed-mode solutions away from the secondary bifurcations. (c) 1999 American Institute of Physics. PMID- 12779872 TI - Two-dimensional global manifolds of vector fields. AB - We describe an efficient algorithm for computing two-dimensional stable and unstable manifolds of three-dimensional vector fields. Larger and larger pieces of a manifold are grown until a sufficiently long piece is obtained. This allows one to study manifolds geometrically and obtain important features of dynamical behavior. For illustration, we compute the stable manifold of the origin spiralling into the Lorenz attractor, and an unstable manifold in zeta(3)-model converging to an attracting limit cycle. (c) 1999 American Institute of Physics. PMID- 12779873 TI - Optimal chaos control through reinforcement learning. AB - A general purpose chaos control algorithm based on reinforcement learning is introduced and applied to the stabilization of unstable periodic orbits in various chaotic systems and to the targeting problem. The algorithm does not require any information about the dynamical system nor about the location of periodic orbits. Numerical tests demonstrate good and fast performance under noisy and nonstationary conditions. (c) 1999 American Institute of Physics. PMID- 12779874 TI - Obtaining functional form for chaotic time series evolution using genetic algorithm. AB - A genetic algorithm (GA) based strategy is presented for deducing an exact or near-exact functional form from a time series. The GA formalism proposed here utilizes (i) the "postfix" representation with a view to reduce the procedural complexities and (ii) the "elitist mating" scheme to produce fitter offspring strings. The GA procedure is exemplified by considering chaotic time series of the well-known logistic, Henon and universal maps. The GA correctly recovers the underlying functional forms for the respective time series. Measurements from a number of finite-dimensional physical, biological, and other systems often give rise to complex time series and the presented methodology should prove useful in obtaining functional forms describing accurately the evolution of the time series. (c) 1999 American Institute of Physics. PMID- 12779875 TI - A coupled ordinary differential equation lattice model for the simulation of epileptic seizures. AB - A coupled ordinary differential equation lattice model for the CA3 region of the hippocampus (a common location of the epileptic focus) is developed. This model consists of a hexagonal lattice of nodes, each describing a subnetwork consisting of a group of prototypical excitatory pyramidal cells and a group of prototypical inhibitory interneurons connected via on/off excitatory and inhibitory synapses. The nodes communicate using simple rules to simulate the diffusion of extracellular potassium. Both the integration time over which a node's trajectory is integrated before the diffusional event is allowed to occur and the level of inhibition in each node were found to be important parameters. Shorter integration times lead to total synchronization of the lattice (similar to synchronous neural activity occurring during a seizure) whereas longer times cause more random spatiotemporal behavior. Moderately diminished levels of inhibition lead to simple nodal oscillatory behavior. It is postulated that both the lack of inhibition and an alteration in conduction time may be necessary for the development of a behaviorally manifest seizure. (c) 1999 American Institute of Physics. PMID- 12779876 TI - Symbolic dynamics of jejunal motility in the irritable bowel. AB - Different studies of the irritable bowel syndrome (IBS) by conventional analysis of jejunal motility report conflicting results. Therefore, our aim is to quantify the jejunal contraction activity by symbolic dynamics in order to discriminate between IBS and control subjects. Contraction amplitudes during fasting motility (phase II) are analyzed for 30 IBS and 30 healthy subjects. On the basis of a particular scale-independent discretization of the contraction amplitudes with respect to the median, IBS patients are characterized by increased block entropy as well as increased mean contraction amplitude. In a further more elementary level of analysis these differences can be reduced to specific contraction patterns within the time series, namely the fact that successive large contraction amplitudes are less ordered in IBS than in controls. These significant differences in jejunal motility may point to an altered control of the gut in IBS, although further studies on a representative number of patients have to be done for a validation of these findings. (c) 1999 American Institute of Physics. PMID- 12779877 TI - Topological analysis of chaos in neural spike train bursts. AB - We show how a topological model which describes the stretching and squeezing mechanisms responsible for creating chaotic behavior can be extracted from the neural spike train data. The mechanism we have identified is the same one ("gateau roule," or jelly-roll) which has previously been identified in the Duffing oscillator [Gilmore and McCallum, Phys. Rev. E 51, 935 (1995)] and in a YAG laser [Boulant et al., Phys. Rev. E 55, 5082 (1997)]. (c) 1999 American Institute of Physics. PMID- 12779878 TI - Global analysis of periodic orbit bifurcations in coupled Morse oscillator systems: time-reversal symmetry, permutational representations and codimension-2 collisions. AB - In this paper we study periodic orbit bifurcation sequences in a system of two coupled Morse oscillators. Time-reversal symmetry is exploited to determine periodic orbits by iteration of symmetry lines. The permutational representation of Tsuchiya and Jaffe is employed to analyze periodic orbit configurations on the symmetry lines. Local pruning rules are formulated, and a global analysis of possible bifurcation sequences of symmetric periodic orbits is made. Analysis of periodic orbit bifurcations on symmetry lines determines bifurcation sequences, together with periodic orbit periodicities and stabilities. The correlation between certain bifurcations is explained. The passage from an integrable limit to nointegrability is marked by the appearance of tangent bifurcations; our global analysis reveals the origin of these ubiquitous tangencies. For period-1 orbits, tangencies appear by a simple disconnection mechanism. For higher period orbits, a different mechanism involving 2-parameter collisions of bifurcations is found. (c) 1999 American Institute of Physics. PMID- 12779879 TI - Dynamics of a gravitational billiard with a hyperbolic lower boundary. AB - Gravitational billiards provide a simple method for the illustration of the dynamics of Hamiltonian systems. Here we examine a new billiard system with two parameters, which exhibits, in two limiting cases, the behaviors of two previously studied one-parameter systems, namely the wedge and parabolic billiard. The billiard consists of a point mass moving in two dimensions under the influence of a constant gravitational field with a hyperbolic lower boundary. An iterative mapping between successive collisions with the lower boundary is derived analytically. The behavior of the system during transformation from the wedge to the parabola is investigated for a few specific cases. It is surprising that the nature of the transformation depends strongly on the parameter values. (c) 1999 American Institute of Physics. PMID- 12779880 TI - Lyapunov exponents, dual Lyapunov exponents, and multifractal analysis. AB - It is shown that the multifractal property is shared by both Lyapunov exponents and dual Lyapunov exponents related to scaling functions of one-dimensional expanding folding maps. This reveals in a quantitative way the complexity of the dynamics determined by such maps. (c) 1999 American Institute of Physics. PMID- 12779881 TI - Calculations of periodic orbits: The monodromy method and application to regularized systems. AB - We describe a numerical method for calculating periodic orbits, which is a generalization of the monodromy method by Baranger et al. to the case of an arbitrary autonomous dynamical system. Two variants of the method are developed, using the midpoint and the Runge-Kutta discretization of equations of motion, respectively. Particularly, we adapt the first variant for calculating periodic orbits of Hamiltonian systems when the period or the energy is given a priori. Finally, we consider the application of the monodromy method to the case of regularized mechanical systems and demonstrate the use by two examples. (c) 1999 American Institute of Physics. PMID- 12779882 TI - Detecting dynamical nonstationarity in time series data. AB - Nonlinear time series analysis is becoming an ever more powerful tool to explore complex phenomena and uncover underlying patterns from irregular data recorded from experiments. However, the existence of dynamical nonstationarity in time series data causes many results of such analysis to be questionable and inconclusive. It is increasingly recognized that detecting dynamical nonstationarity is a crucial precursor to data analysis. In this paper, we present a test procedure to detect dynamical nonstationarity by directly inspecting the dependence of nonlinear statistical distributions on absolute time along a trajectory in phase space. We test this method using a broad range of data, chaotic, stochastic and power-law noise, both computer-generated and observed, and show that it provides a reliable test method in analyzing experimental data. (c) 1999 American Institute of Physics. PMID- 12779883 TI - Transport in finite size systems: An exit time approach. AB - In the framework of chaotic scattering we analyze passive tracer transport in finite systems. In particular, we study models with open streamlines and a finite number of recirculation zones. In the nontrivial case with a small number of recirculation zones a description by means of asymptotic quantities (such as the eddy diffusivity) is not appropriate. The nonasymptotic properties of dispersion are characterized by means of the exit time statistics, which shows strong sensitivity on initial conditions. This yields a probability distribution function with long tails, making impossible a characterization in terms of a unique typical exit time. (c) 1999 American Institute of Physics. PMID- 12779884 TI - Threshold, excitability and isochrones in the Bonhoeffer-van der Pol system. AB - Some new insight is obtained for the structure of the Bonhoeffer-van der Pol system. The problems of excitability and threshold are discussed for all three types of the system classified according to the existing attractors: a focus only, a limit cycle only and a limit cycle together with a focus. These problems can be treated by the T-repellers and the T-attractors of the system which are mutually reciprocal under time inversion. The threshold depends on the structure of the T-repeller (unstable part of integral manifold). This structure is then used to understand the behavior and the properties of the two different types of isochrones: Winfree isochrones (W-isochrones) and regular isochrones. Winfree's description of a W-isochrone is extended to excitable systems. Both W-isochrones and regular isochrones are calculated for the Bonhoeffer-van der Pol system in its limit cycle and excitable regimes. The important role of the T-repeller as an asymptotic limit for both types of isochrones is manifested. (c) 1999 American Institute of Physics. PMID- 12779885 TI - An expansion of system with time delayed feedback control into spatio-temporal state space. AB - Time delayed feedback control is well known as an effective continuous control method for stabilizing the unstable periodic orbit embedded in chaotic attractors. As for the system with time delay, the solution is in a function space and shows characteristics governed by an infinite dimension. Therefore it is difficult to understand the system behavior analytically. In this paper, it is shown that, when the state space for the system with time delay is expanded into the spatio-temporal state space, the solution propagates in the space as a wave theoretically and numerically. The dynamic behavior experimentally obtained in the sinusoidally excited magneto-elastic beam system under time delayed feedback control is also discussed by the transformation into the expanded spatio-temporal state space. (c) 1999 American Institute of Physics. PMID- 12779886 TI - Traveling waves and chaotic properties in cellular automata. AB - Traveling wave solutions of cellular automata (CA) with two states and nearest neighbors interaction on one-dimensional (1-D) infinite lattice are computed. Space and time periods and the number of distinct waves have been computed for all representative rules, and each velocity ranging from 2 to 22. This computation shows a difference between spatially extended systems, generating only temporal chaos and those producing as well spatial complexity. In the first case wavelengths are simply related to the velocity of propagation and the dispersivity is an affine function, while in the second case (which coincides with Wolfram class 3), the dispersivity is multiform and its dependence on the velocities is highly random and discontinuous. This property is typical of space time chaos in CA. (c) 1999 American Institute of Physics. PMID- 12779887 TI - Anomalous spatio-temporal chaos in a two-dimensional system of nonlocally coupled oscillators. AB - A two-dimensional system of nonlocally coupled complex Ginzburg-Landau oscillators is investigated numerically for the first time. As previously shown for the one-dimensional case, this two-dimensional system exhibits anomalous spatio-temporal chaos characterized by power-law spatial correlations. In this chaotic regime, the amplitude difference between neighboring elements displays temporal noisy on-off intermittency. The system is also spatially intermittent in this regime, as revealed by multiscaling analysis: The amplitude field is multiaffine and the difference field is multifractal. Correspondingly, the probability distribution function of the measure defined for each field is strongly non-Gaussian, exhibiting scale-dependent deviations in the tail due to intermittency. (c) 1999 American Institute of Physics. PMID- 12779888 TI - Chaotic mixing of granular material in slowly rotating containers as a discrete mapping. AB - Chaotic mixing of granular material in a two-dimensional slowly rotating noncircular container in the absence of granular diffusivity is studied analytically and numerically as a discrete mapping. The noncircularity of a drum produces a time periodic disturbance and chaotization of the flow field. The location of the fixed points of the mapping and the separatrices of the hyperbolic points are determined in a closed analytical form.(c) 1999 American Institute of Physics. PMID- 12779889 TI - Hysteresis and bistability in the direct transition from 1:1 to 2:1 rhythm in periodically driven single ventricular cells. AB - The transmembrane potential of a single quiescent cell isolated from rabbit ventricular muscle was recorded using a suction electrode in whole-cell recording mode. The cell was then driven with a periodic train of current pulses injected into the cell through the same recording electrode. When the interpulse interval or basic cycle length (BCL) was sufficiently long, 1:1 rhythm resulted, with each stimulus pulse producing an action potential. Gradual decrease in BCL invariably resulted in loss of 1:1 synchronization at some point. When the pulse amplitude was set to a fixed low level and BCL gradually decreased, N+1:N rhythms (N>/=2) reminiscent of clinically observed Wenckebach rhythms were seen. Further decrease in BCL then yielded a 2:1 rhythm. In contrast, when the pulse amplitude was set to a fixed high level, a period-doubled 2:2 rhythm resembling alternans rhythm was seen before a 2:1 rhythm occurred. With the pulse amplitude set to an intermediate level (i.e., to a level between those at which Wenckebach and alternans rhythms were seen), there was a direct transition from 1:1 to 2:1 rhythm as the BCL was decreased: Wenckebach and alternans rhythms were not seen. When at that point the BCL was increased, the transition back to 1:1 rhythm occurred at a longer BCL than that at which the {1:1-->2:1} transition had initially occurred, demonstrating hysteresis. With the BCL set to a value within the hysteresis range, injection of a single well-timed extrastimulus converted 1:1 rhythm into 2:1 rhythm or vice versa, providing incontrovertible evidence of bistability (the coexistence of two different periodic rhythms at a fixed set of stimulation parameters). Hysteresis between 1:1 and 2:1 rhythms was also seen when the stimulus amplitude, rather than the BCL, was changed. Simulations using numerical integration of an ionic model of a single ventricular cell formulated as a nonlinear system of differential equations provided results that were very similar to those found in the experiments. The steady-state action potential duration restitution curve, which is a plot of the duration of the action potential during 1:1 rhythm as a function of the recovery time or diastolic interval immediately preceding that action potential, was determined. Iteration of a finite-difference equation derived using the restitution curve predicted the direct {1:1<-->2:1} transition, as well as bistability, in both the experimental and modeling work. However, prediction of the action potential duration during 2:1 rhythm was not as accurate in the experiments as in the model. Finally, we point out a few implications of our findings for cardiac arrhythmias (e.g., Mobitz type II block, ischemic alternans). (c) 1999 American Institute of Physics. PMID- 12779890 TI - Stochastic webs and their applications. AB - The conditions for the appearance of a stochastic web in degenerate dynamic systems and typical physical problems that lead to such a web are analyzed. Examples of webs are considered, as well as their symmetry, width, and structural changes. A description is given of a change in the diffusion dynamics along the web channels as a function of the number of the degrees of freedom and the phenomenon of stochastic percolation is discussed. PMID- 12779891 TI - Cardiac arrhythmias and circle maps-A classical problem. AB - The periodic forcing of nonlinear oscillations can often be cast as a problem involving self-maps of the circle. Consideration of the effects of changes in the frequency and amplitude of the periodic forcing leads to a problem involving the bifurcations of circle maps in a two-dimensional parameter space. The global bifurcations in this two-dimensional parameter space is described for periodic forcing of several simple theoretical models of nonlinear oscillations. As was originally recognized by Arnold, one motivation for the formulation of these models is their connection with theoretical models of cardiac arrhythmias originating from the competition and interaction between two pacemakers for the control of the heart. PMID- 12779892 TI - Cardiac arrhythmias and circle mappings(a)). AB - The description of the Gelfand-Tsetlin model is presented as excerpts from the author's diploma dissertation on the mappings of a circle to itself. PMID- 12779893 TI - An approach to renormalization on the n-torus. AB - The coding theory of rotations (by inspecting closely their relation to flows) and the continued fractions algorithm (by considering even two-coloring of the integers with a given proportion of, say, blue and red) are revisited. Then, even n-coloring of the integers is defined. This allows one to code rotations on the (n-1)-torus by considering linear flows on the n-torus and yields a simple geometric approach to renormalization on tori by first return maps on the coding regions. PMID- 12779894 TI - Experimental measurement of chaotic attractors in solid mechanics(a)). AB - In this paper a review is given of experimental techniques in chaotic dynamics of solid mechanical systems based on modern ideas of nonlinear dynamics. These methods include Poincare maps, double Poincare sections, symbol dynamics, and fractal dimension. The physical problems discussed include nonlinear elastic beams, forced motion of a string, flow-induced vibration of a rod, forced motions of a magnetic pendulum, and rigid body dynamics of a magnet and high-temperature superconductor. PMID- 12779895 TI - Probability phenomena due to separatrix crossing. AB - The effect of a small or slow perturbation on a Hamiltonian system with one degree of freedom is considered. It is assumed that the phase portrait ("phase plane") of the unperturbed system is divided by separatrices into several regions and that under the action of the perturbations phase points can cross these separatrices. The probabilistic phenomena are described that arise due to these separatrix crossings, including the scattering of trajectories, random jumps in the values of adiabatic invariants, and adiabatic chaos. These phenomena occur both in idealized problems in classical mechanics and in real physical systems in planetary science and plasma physics contexts. PMID- 12779896 TI - Chaos in a multimode solid-state laser system. AB - When a nonlinear crystal is placed within a multimode solid-state laser cavity, deterministic fluctuations are induced in the output intensity. In this paper, the results of our studies of the intensity noise in a diode pumped, intracavity frequency doubled Nd:YAG (neodymium doped yttrium aluminum garnet) laser will be presented. First, a novel technique to eliminate these fluctuations is described. Second, the observation of antiphase states in the laser output is discussed. These states are characterized by a cyclic periodic pulsing of the individual longitudinal mode intensities. Finally, the statistical properties of chaotic intensity fluctuations are characterized. It is be demonstrated that it is possible to accurately model the laser dynamics by a system of globally coupled, nonlinear oscillators. PMID- 12779897 TI - Symbol dynamic maps of spatial-temporal chaotic vibrations in a string of impact oscillators. AB - Spatially complex, temporally chaotic dynamics of N-coupled impact oscillators connected by a string are studied experimentally using a discrete measure of the motion for each of the masses. For N=8, a binary assignment of symbols, corresponding to whether or not the masses impact an amplitude constraint, is used to code the spatial pattern as a binary number and to store its change in time in a computer. A spatial pattern return map is then used to observe the change in spatial patterns with time. Bifurcations in spatial impact patterns are observed in this experiment. An entropy measure is also used to characterize the dynamics. Numerical simulation shows behavior similar to the experimental system. PMID- 12779898 TI - Diffusion-induced instability in chemically reacting systems: Steady-state multiplicity, oscillation, and chaos. AB - The dynamical behavior of two coupled cells or reactors is described. The cells are coupled by diffusion, e.g., through a semipermeable membrane, and the chemical reactions and initial or feed concentrations of all species are the same in the two cells. Each cell has only a single stable steady state in the absence of coupling, and the coupled system may exhibit multiple steady states, periodic oscillation, or chaos. The attractors of the coupled system may be either homogeneous (the two cells have equal concentrations) or inhomogeneous. Three two variable kinetic models are examined: the Brusselator, a model of the chlorine dioxide-iodine-malonic acid reaction, and the Degn-Harrison model. The dynamical behavior of the coupled system is determined by the nonlinearities in the uncoupled subsystems and by two ratios, that of the diffusion constants of the two species and that of the area of the membrane to the product of the membrane thickness and the volume of a cell. PMID- 12779899 TI - Identifying coherent structures in nonlinear wave propagation. AB - Nonlinear wave phenomena are often characterized by the appearance of "solitary wave coherent structures" traveling at speeds determined by their amplitudes and morphologies. Assuming that time intervals exist in which these structures are essentially noninteracting, a method for identifying the number of independent features and their respective speeds is proposed and developed. The method is illustrated with a variety of increasingly realistic specific applications, beginning with a simple nonlinear but analytically tractable Gaussian model, continuing with (numerically generated) data describing multisoliton solutions to the Korteweg-de Vries equation, and concluding with (numerical) data from a realistic simulation of nonlinear wave interactions in plasma turbulence. These studies reveal both strengths and limitations of the method in its present incarnation and suggest topics for future investigations. PMID- 12779900 TI - A theory of quantum diffusion localization. AB - The quantum localization of chaotically diffusive classical motion is reviewed, using the kicked rotator as a simple but instructive example. The specific quantum steady state, which results from statistical relaxation in the discrete spectrum, is described in some detail. A new phenomenological theory of quantum dynamical relaxation is presented and compared with the previously existing theory. PMID- 12779901 TI - Chaotic scattering, unstable periodic orbits, and fluctuations in quantum transport. AB - The field of quantum chaos has recently focused attention on the quantum description of chaotic scattering processes. The new physical intuition, analytical methods, and numerical tools developed in the study of the quantum behavior of classically chaotic bound systems, like quantum billiards or atoms in strong fields, has led to exciting new predictions for the scattering of electromagnetic waves in curved waveguides, electrons in mesoscopic wires, and atoms off molecules. After a brief review of recent progress in the field of quantum chaos, this paper focuses on specific results relating to ballistic electron transport in small, mesoscopic devices. Several specific geometries are suggested for experimental studies of this "game" of quantum pinball with explicit predictions for the fluctuations in the electrical conductivity as functions of the electron Fermi energy and of an applied magnetic field. PMID- 12779902 TI - Near-classical noise enhancement of microwave ionization of Rydberg atoms. AB - The ionization of the highly excited hydrogen atom in a strong external microwave field is a classically chaotic, near-classical quantum system for microwave frequencies somewhat below the initial Kepler electron orbit frequency. The addition of microwave noise is found to reduce the sinewave microwave field needed for ionization, modifying the near-classical fast process responsible for the microwave energy absorption. A classical numerical calculation based upon a many-frequency model of the noise qualitatively reproduces the observed noise enhancement. PMID- 12779903 TI - Spectral and stability aspects of quantum chaos. AB - The relation between the spectrum of a generalized quasienergy operator and the stability of quantum systems driven by quasiperiodic time-dependent forces is discussed. PMID- 12779904 TI - Limiting probability distribution for a random walk with topological constraints. AB - The joint limiting probability distribution is studied for the two-dimensional random walk with topological constraints, omega(2ns), on Z(2) lattice, where 2n is its total length and (0X with small "traps" Y( varepsilon ) subset, dbl equals X (diam(Y( varepsilon )) infinity conditional probabilities that f(n+1)x in X?Y( varepsilon ) if x,fx,.,f(nx) in X?Y( varepsilon ) and the point x is chosen at random. Also proven is the convergence of &mgr;( varepsilon ) to smooth f-invariant measures as varepsilon -->0. By means of this construction, the numerical phenomenon of the convergence of histograms of trajectories of maps with marginal singularities to densities of nonfinite smooth invariant measures in the computer modeling was investigated. PMID- 12779933 TI - Rate processes in nonlinear optical dynamics with many attractors. AB - Kramers' 1940 paper and its successive elaborations have extensively explored the transition rate between two stable situations, that is, in the language of system dynamics, the transition between the basins of attraction of two stable fixed point attractors. In a nonequilibrium system some of the above conditions may be violated, either because one of the two fixed points is unstable, as in the case of transient phenomena, or because both fixed points are unstable, as in the case of heteroclinic chaos, or because the attractors are more complex than fixed points, as in a chaotic dynamics where two or more strange attractors coexist. Furthermore, there is recent experimental evidence of space-time complexity consisting in the alternate or simultaneous oscillation of many modes, each one with its own (possibly chaotic) dynamics. In all the above cases, coexistence of many alternative paths implies a choice, either due to noise or self-triggered by the same interacting degrees of freedom. A review of the above phenomena in the case of nonequilibrium optical systems is here presented, with the aim of stimulating theoretical investigation on these novel rate processes. PMID- 12779934 TI - A history of chemical oscillations and waves. AB - The history of the discovery and study of chemical oscillations and waves is presented from the very first accidental observations up to the systematic design of chemical oscillators. Special emphasis is devoted to the long-term debate over the possibility of pure chemical oscillations, i.e., concentration oscillations in homogeneous closed systems. PMID- 12779935 TI - Oscillations and chaos in CO+O(2) combustion. AB - The gas-phase reaction between carbon monoxide and oxygen (in the presence of small amounts of hydrogen) shows bistability and oscillatory behavior. Typically, the oscillatory ignition has a period-1 relaxation waveform. The limit cycle is born at a saddle-node loop and terminates via a supercritical Hopf bifurcation. For a mean residence time of 8 s there is a period-doubling to a period-2 solution followed by period-halving to quasisinusoidal period-1 oscillations. At longer residence times, more period-doublings forming a full cascade to chaos with subsequent periodic windows are observed. The chaotic attractor has an underlying single-humped next maximum map. PMID- 12779936 TI - Fractal analysis of size effects and surface morphology effects in catalysis and electrocatalysis. AB - Fractal geometry tools are used in order to analyze several related problems in surface science, catalysis, and electrocatalysis. The effects of complex morphologies of adsorbents, catalysts, and electrodes on various molecular processes with these materials are determined both theoretically and experimentally. It is shown that fractal geometry provides a convenient and natural tool for the elucidation of geometry-performance relations in heterogeneous chemistry. Issues covered are particle size effects in physisorption and chemisorption; morphology effects on a variety of catalytic processes with unsupported catalysts (including coal liquefaction, alkene polymerizations, oxidations, dehydrogenations, and esterifications); surface accessibility effects on molecular interactions in an Eley-Rideal mechanism; surface patterning effects on concentration profiles near the surface; and electrode-morphology effects on a variety of electrochemical and electrocatalytic processes. The domains of applicability of the fractal approach to these problems is discussed. PMID- 12779937 TI - Transition to chemical turbulence. AB - Experiments have been conducted on Turing-type chemical spatial patterns and their variants in a quasi-two-dimensional open spatial reactor with a chlorite iodide-malonic acid reaction. A variety of stationary spatial structures hexagons, stripes, and mixed states-were observed, and transitions to these states were studied. For conditions beyond those corresponding to the emergence of patterns, a transition was observed from stationary spatial patterns to chemical turbulence, which is marked by a continuous motion of the pattern within a domain and of the grain boundaries between domains. The transition to chemical turbulence was analyzed by measuring the correlation length, the average pattern speed, and the total length of the domain boundaries. The emergence of chemical turbulence is accompanied by a large increase in the defects in the pattern, which suggests that this is an example of defect-mediated turbulence. PMID- 12779938 TI - Vortex dynamics in oscillatory chemical systems. AB - Vortex core dynamics is studied in the Brusselator both near to and far from the Hopf bifurcation line for random and pair initial conditions. Extensive simulations are carried out for a pair of counter-rotating vortices close to the Hopf bifurcation line. Provided the vortices are not so far apart that wave-front annihilation produces strong gradients between their centers, the simulation results compare favorably with theories based on the complex Ginzburg-Landau equation. Far from the Hopf line the vortex core dynamics changes character and phenomena such as periodic motion of the vortex centers arise. PMID- 12779939 TI - Transverse coupling of chemical waves. AB - The transverse coupling of chemical waves is investigated using a model scheme for excitable media. Chemical waves supported on the surfaces of a semipermeable membrane couple via diffusion through the membrane, resulting in new types of spatiotemporal behavior. The model studies show that spontaneous wave sources may develop from interacting planar waves, giving rise to a complex sequence of patterns accessible only by perturbation. Coupled circular waves result in the spontaneous formation of spiral waves, which subsequently develop patterns in distinct domains with characteristic features. The long time entrainment behavior of coupled spiral waves reveals regions of 1:2 phase locking. PMID- 12779940 TI - Bifurcation structures of periodically forced oscillators. AB - A theoretical investigation of bifurcation structures of periodically forced oscillators is presented. In the plane of forcing frequency and amplitude, subharmonic entrainment occurs in v-shaped (Arnol'd) tongues, or entrainment bands, for small forcing amplitudes. These tongues terminate at higher forcing amplitudes. Between these two limits, individual tongues fit together to form a global bifurcation structure. The regime in which the forcing amplitude is much smaller than the amplitude of the limit cycle is first examined. Using the method of multiple time scales, expressions for solutions on the invariant torus, widths of Arnol'd tongues, and Liapunov exponents of periodic orbits are derived. Next, the regime of moderate to large forcing amplitudes is examined through studying a periodically forced Hopf bifurcation. In this case the forcing amplitude and the amplitude of the limit cycle can be of the same order of magnitude. From a study of the normal forms for this case, it is shown how Arnol'd tongues terminate and how complicated bifurcation structures are associated with strong resonances. Aspects of model and experimental chemical systems that show some of the phenomena predicted from the above theoretical results are mentioned. PMID- 12779941 TI - Geometric phases in dissipative systems. AB - It is shown that a phenomenon analogous to the geometric phase shifts of Berry and Hannay occurs for dissipative oscillatory systems and can be detected in numerical simulations of chemical oscillators. The approach herein to the theory of geometric phases begins with a study of simple first-order differential equations on the circle (circle dynamics). It is shown how more complicated systems exhibit geometric phases through reduction to a circle dynamics. In this way, the various manifestations of the phenomenon are seen from a single unified perspective. The results are illustrated in numerical experiments on several model systems ranging from analytically solvable, but contrived, to realistic models of chemical oscillators. PMID- 12779943 TI - Parameter dependence of stochastic layers in a quasicrystalline web. AB - Stochastic web maps with approximate quasicrystalline symmetry possess an infinite number of inequivalent fixed points embedded in stochastic layers of varying thickness. In this investigation exploratory steps are taken toward a systematic numerical determination of the widths of the stochastic layers as a function of the web map's control parameter. The study concentrates on a particular stochastic layer in the approximately fivefold symmetric web. Computer graphics and a simple stretching-and-folding criterion provide a coarse view, which is supplemented at finer scales by Greene's residue method. The exact reflection symmetries of invariant sets, as well as a five-dimensional representation of the map, are exploited to improve numerical precision. As the control parameter varies, one finds not only variations expected from island chain structures, but also larger-scale oscillations whose origin is not understood. PMID- 12779942 TI - Channeling and percolation in two-dimensional chaotic dynamics. AB - The Hamiltonian dynamics of a particle moving in a nearly periodic two dimensional (2-D) potential of square symmetry is analyzed. The particle undergoes two types of unbounded stochastic or random walks in such a system: a quasi-1-D motion (a "stochastic channeling") and a 2-D motion which results from a sort of stochastic percolation. A scenario for the onset of this stochastic percolation is analyzed. The threshold energy for percolation is found as a function of the perturbation parameter. Each type of random walk has the property of intermittency. The particle transport is anomalous in certain energy intervals. PMID- 12779944 TI - Periodic orbit theory in classical and quantum mechanics. PMID- 12779945 TI - On dynamical zeta function. AB - The dynamical zeta function is usually defined as an infinite (and divergent) product over all primitive periodic orbits. It is possible to show that as variant Planck's over 2pi -->0 it can be represented as det(1-T), where the operator T(q,q') defines the semiclassical Poincare map. Here, certain consequences of this representation for chaotic systems are discussed. In particular, it is shown that the zeta function can be expressed through a subset of specially selected orbits, the error of this approximation being small as variant Planck's over 2pi -->0. Assuming that the chosen Poincare surface of section is divided into small cells of phase-space area of 2pi variant Planck's over 2pi, these trajectories are uniquely characterized by the requirement that they never go twice through the same cell. PMID- 12779946 TI - The semiclassical functional equation. AB - A semiclassical analog of the functional equation for the Riemann zeta function is considered. In the case of the zeta function itself, this equation forms the basis for a finite approximation to the Dirichlet series, known as the approximate functional equation. In the same way, the semiclassical functional equation can be shown to give rise to a finite approximation to the semiclassical representation of the quantum spectral determinant as a sum over classical pseudo orbits. This finite approximation has been called the Riemann-Siegel look-alike formula. The formal nature of the derivation of this result is discussed and the fact that it appears to imply a remarkable relationship between long and short pseudo-orbits is shown. PMID- 12779947 TI - The semiclassical helium atom. AB - Recent progress in the semiclassical description of two-electron atoms is reported herein. It is shown that the classical dynamics for the helium atom is of mixed phase space structure, i.e., regular and chaotic motion coexists. Semiclassically, both types of motion require separate treatment. Stability islands are quantized via a torus-quantization-type procedure, whereas a periodic orbit cycle expansion approach accounts for the states associated with hyperbolic electron pair motion. The results are compared with highly accurate ab initio quantum calculations, most of which are reported here for the first time. The results are discussed with an emphasis on previous interpretations of doubly excited electron states PMID- 12779948 TI - Applications of periodic-orbit theory. AB - The periodic-orbit theory of Gutzwiller is applied in various ways by using generalized periodic-orbit sum rules. Numerical evaluations are carried out for the hyperbola billiard, a strongly chaotic system. The most efficient semiclassical determination of quantum energies is achieved by a quantization condition, which is formulated in terms of a zeta function by using a functional equation. PMID- 12779949 TI - Semiclassical mechanics of bound chaotic potentials. AB - Semiclassical methods for determining quantum eigenvalues in chaotic systems are discussed. A recent calculation for an open scattering system with Axiom-A properties serves as a starting point for the discussion. How deviation from Axiom-A properties, such as intermittency and occurrence of small stability islands, normally arise in bound Hamiltonian systems, and how these deviations complicate the calculation of semiclassical eigenvalues are demonstrated. It is also stressed that since such deviations are typical of bound Hamiltonian systems, they might be of crucial importance for the statistical properties of the energy levels. PMID- 12779950 TI - Quantization of chaotic systems. AB - Starting from the semiclassical dynamical zeta function for chaotic Hamiltonian systems we use a combination of the cycle expansion method and a functional equation to obtain highly excited semiclassical eigenvalues. The power of this method is demonstrated for the anisotropic Kepler problem, a strongly chaotic system with good symbolic dynamics. An application of the transfer matrix approach of Bogomolny is presented leading to a significant reduction of the classical input and to comparable accuracy for the calculated eigenvalues. PMID- 12779951 TI - Periodic orbit quantization of the anisotropic Kepler problem. AB - The periodic orbit quantization on the anisotropic Kepler problem is tested. By computing the stability and action of some 2000 of the shortest periodic orbits, the eigenvalue spectrum of the anisotropic Kepler problem is calculated. The aim is to test the following claims for calculating the quantum spectrum of classically chaotic systems: (1) Curvature expansions of quantum mechanical zeta functions offer the best semiclassical estimates; (2) the real part of the cycle expansions of quantum mechanical zeta functions cut at appropriate cycle length offer the best estimates; (3) cycle expansions are superfluous; and (4) only a small subset of cycles (irreducible cycles) suffices for good estimates for the eigenvalues. No evidence is found to support any of the four claims. PMID- 12779952 TI - Pruning of orbits in four-disk and hyperbola billiards. AB - It is shown that for the four-disk system and the hyperbola billiard it is possible to construct a new symbolic plane preserving the orientation existing in the dynamical space. Physical orbits are mapped into the topological well-ordered plane and it is shown that the forbidden and allowed orbits are separated by a monotone pruning front. PMID- 12779953 TI - Validity of the semiclassical periodic orbit approximation in the two- and three disk problems. AB - The high-lying resonances in the quantum mechanical scattering problem of a point particle from two or three equally sized (and spaced) circular hard disks in the two-dimensional plane are predicted quite well by the classical cycle expansion. There are, however, noticeable deviations for the lowest resonances. Therefore, the leading corrections from creeping paths to the cycle expansion in the two disk scattering problem are constructed. Generalizations to the three-disk problem are indicated. The size of the corrections are estimated. They are shown to be too small to account for the deviations mentioned above. Finally, arguments are given that, for the two- and three-disk problem, the semiclassical predictions of the low-lying resonance poles are bound to fail. PMID- 12779954 TI - Investigation of the Lorentz gas in terms of periodic orbits. AB - The diffusion constant and the Lyapunov exponent for the spatially periodic Lorentz gas are evaluated numerically in terms of periodic orbits. A symbolic description of the dynamics reduced to a fundamental domain is used to generate the shortest periodic orbits. Applied to a dilute Lorentz gas with finite horizon, the theory works well, but for the dense Lorentz gas the convergence is hampered by the strong pruning of the admissible orbits. PMID- 12779955 TI - Cycle expansion for the Lyapunov exponent of a product of random matrices. AB - Using cycle expansion for the thermodynamic zeta function, a formula is derived for the Lyapunov exponent of a product of random hyperbolic matrices chosen from a discrete set. This allows for an accurate numerical solution of the Ising model in one dimension with quenched disorder. The formula is compared with weak disorder expansions and with the microcanonical approximation and shown to apply to matrices with degenerate eigenvalues. PMID- 12779956 TI - Anatomy of the trace formula for the baker's map. AB - The structure of the trace formula for quantum maps on a compact phase space is analyzed. An explicit expression for the functional determinant in terms of a finite number of traces is derived which is algebraic and independent of any approximation. For the specific case of the baker's map, its simple structure allows the implementation of a symbolic decomposition of the propagator which is exact and which has the structure of usual semiclassical formulas. The method allows the testing of the accuracy of the individual contribution of each periodic orbit to the functional determinant. PMID- 12779957 TI - Time domain approach to semiclassical dynamics: Breaking the log time barrier. AB - The presence of chaos in the classical dynamics is not necessarily the destructive element it was thought to be for semiclassical approximations in the time domain. The method of calculating the semiclassical propagation of initial states and correlation functions for nonlinear and chaotic dynamics is shown, and the excellent accuracy is noted for rather long times. The breakdown timescale is much longer than the infamous "log time" for the cases investigated here. PMID- 12779958 TI - Chaotic spectroscopy. AB - The spectra of quantized chaotic billiards from the point of view of scattering theory are discussed. It is shown how the spectral and resonance density functions both fluctuate about a common mean. A semiclassical treatment explains this in terms of classical scattering trajectories and periodic orbits of the Poincare scattering map. It is shown that this formalism provides an alternative derivation and a new interpretation of Gutzwiller's periodic orbits sum for the spectral density. Moreover, it is a convenient starting point for a derivation of a Riemann-Siegel "look alike" expression for the secular equation in terms of periodic orbits of finite length. PMID- 12779959 TI - Topological aspects of quantum chaos. AB - Quantized classically chaotic maps on a toroidal two-dimensional phase space are studied. A discrete, topological criterion for phase-space localization is presented. To each eigenfunction is associated an integer, analogous to a quantized Hall conductivity, which tests the way the eigenfunction explores the phase space as some boundary conditions are changed. The correspondence between delocalization and chaotic classical dynamics is discussed, as well as the role of degeneracies of the eigenspectrum in the transition from localized to delocalized states. The general results are illustrated with a particular model. PMID- 12779960 TI - Experimental evidence for the influence of "scars" in hydrogen atoms driven by strong microwave fields. AB - A preliminary analysis is reported of new experimental data for the microwave ionization of excited hydrogen atoms with each of the principal quantum numbers n(0) between 50 and 79. Data are shown for a linearly polarized field at each of the frequencies 26.43, 30.36, and 36.02 GHz produced in a resonant cavity. For each frequency certain n(0) values are more stable against ionization; this is called "local stability." The data demonstrate classical scaling of these cases of local stability including certain cases that have been explained classically and others that are nonclassical. The data strongly support a theoretical explanation for nonclassical local stability being due to the influence of "scarred" wave functions. PMID- 12779961 TI - Winding number formula for Maslov indices. AB - A formula for the Maslov index of closed curves on Lagrangian manifolds is derived. The index is expressed as the number of times the plane tangent to the curve winds around it. Applications include unstable periodic orbits, in which case the Lagrangian manifolds are the stable or unstable manifolds of the orbits, and cycles on the invariant tori of integrable systems, in which case the manifolds are the tori themselves. PMID- 12779962 TI - Phase integral theory, coupled wave equations, and mode conversion. AB - Phase integral or WKB theory is applied to multicomponent wave equations, i.e., wave equations in which the wave field is a vector, spinor, or tensor of some kind. Specific examples of physical interest often have special features that simplify their analysis, when compared with the general theory. The case of coupled channel equations in atomic or molecular scattering theory in the Born Oppenheimer approximation is examined in this context. The problem of mode conversion, also called surface jumping or Landau-Zener-Stuckelberg transitions, is examined in the multidimensional case, and cast into normal form. The group theoretical principles of the normal form transformation are laid out, and shown to involve both the Lorentz group and the symplectic group. PMID- 12779963 TI - Pattern dynamics and heat transport in rotating Rayleigh-Benard convection. AB - Pattern dynamics and heat transport in moderate aspect ratio (Gamma=10) rotating Rayleigh-Benard convection are discussed. Patterns are obtained from shadowgraph visualization of the temperature field. Using 2-D Fourier transform analysis of the patterns, it is demonstrated that organized patterns with one dominant roll orientation produce greater thermal heat transport than do more disordered patterns generated by the Kuppers-Lortz instability and consisting of combinations of different roll orientations. PMID- 12779964 TI - Coupled map models for chaos in extended systems. AB - Coupled maps with conserved quantities are introduced as models for chaos in extended systems. The long-wavelength limit of a simple one-dimensional example is investigated in detail. A Langevin equation for this system is derived, and the applicability of the fluctuation-dissipation theorem is discussed. Several numerical experiments to investigate possible thermodynamic properties are described. PMID- 12779965 TI - Coupling of the relaxation and resonant elements in the autonomous chaotic relaxation oscillator (ACRO). AB - If a harmonic oscillator is embedded in a relaxation oscillator, the resulting system may behave like an autonomous chaotic relaxation oscillator (ACRO). The discharge transient of the relaxation oscillator excites sinusoidal oscillations in the harmonic oscillator and these sinusoids affect when the next discharge occurs. This can lead to chaotic intervals in the oscillator periods. A simple electronic model of the ACRO is studied over a wide range of parameters using numerical, analytic, and experimental techniques. The dynamics of the ACRO is found to be determined by three parameters: (1) tuning, (2) coupling, and (3) damping. Complex, intermittent outputs can always be inhibited by increasing the damping of the harmonic oscillator. For weak damping, strong coupling yields chaotic periods. With weak damping and weak coupling, complex behavior only occurs if the relaxation oscillator is tuned near a resonance of the harmonic oscillator. A new path to chaos, called a disruption bifurcation, is the source for intermittency in the ACRO. This bifurcation occurs when the amplitude of internal resonances is excited to the degree that existing limit cycles are disrupted. PMID- 12779966 TI - The evolution of nonlinear resonant oscillations in closed tubes: Solution of the standard mapping. AB - The concern of this paper is the evolution of small-amplitude resonant oscillations of an inviscid gas in a closed tube. The evolution of the oscillations of the gas generated at and near the fundamental frequency and half the fundamental frequency, where experiment shows that shocks are a feature of the final periodic motion, is examined. The basis for the analysis of this nonlinear initial value, boundary value problem on a semi-infinite strip is the Dissipative Standard Map. Since the purpose is to elucidate how the initial rest state of the gas evolves to the final periodic state, the focus of the analysis is on how a prescribed initial curve is mapped, under the Dissipative Standard Map, onto an invariant curve. The method used is to approximate the Dissipative Standard Map, in the small rate limit, by the partial differential equation appropriate to the resonance in question. PMID- 12779967 TI - Calculations of periodic trajectories for the Henon-Heiles Hamiltonian using the monodromy method. AB - The monodromy method, for calculating classical periodic trajectories, is applied to the famous Henon-Heiles potential, which is invariant under the group D(3). The monodromy method is computationally very efficient and is used to find many families of periodic trajectories, including a number of simple bifurcations from the main families of the Henon-Heiles potential. PMID- 12779968 TI - Nonlinear forecasting and iterated function systems. AB - The theory of dynamical forecasting can be extended to iterated function systems. An unordered set of iterates may be sufficient to construct a simulation of the unknown dynamics. The underlying dynamical system may be nondeterministic: A random element may be allowed in the dynamics, and retrieved by our proposed procedures. A first application of these extensions pertains to the reconstruction of fractal attractors, and allows us to solve the so-called inverse problem in Iterated Functions Systems. PMID- 12779969 TI - Bifurcations and traveling waves in a delayed partial differential equation. AB - Here cell population dynamics in which there is simultaneous proliferation and maturation is considered. The resulting mathematical model is a nonlinear first order partial differential equation for the cell density u(t,x) in which there is retardation in both temporal (t) and maturation variables (x), and contains three parameters. The solution behavior depends on the initial function varphi(x) and a three component parameter vector P=(delta,lambda,r). For strictly positive initial functions, varphi(0) greater, similar 0, there are three homogeneous solutions of biological (i.e., non-negative) importance: a trivial solution u(t) identical with 0, a positive stationary solution u(st), and a time periodic solution u(p)(t). For varphi(0)=0 there are a number of different solution types depending on P: the trivial solution u(t), a spatially inhomogeneous stationary solution u(nh)(x), a spatially homogeneous singular solution u(s), a traveling wave solution u(tw)(t,x), slow traveling waves u(stw)(t,x), and slow traveling chaotic waves u(scw)(t,x). The regions of parameter space in which these solutions exist and are locally stable are delineated and studied. PMID- 12779970 TI - Fractal dimension of steady nonequilibrium flows. AB - The Kaplan-Yorke information dimension of phase-space attractors for two kinds of steady nonequilibrium many-body flows is evaluated. In both cases a set of Newtonian particles is considered which interacts with boundary particles. Time averaged boundary temperatures are imposed by Nose-Hoover thermostat forces. For both kinds of nonequilibrium systems, it is demonstrated numerically that external isothermal boundaries can drive the otherwise purely Newtonian flow onto a multifractal attractor with a phase-space information dimension significantly less than that of the corresponding equilibrium flow. Thus the Gibbs' entropy of such nonequilibrium flows can diverge. PMID- 12779971 TI - Discrete model of a dynamo with diffusion. AB - A model of magnetic field generation by the turbulent motions of a highly conducting fluid is constructed. It is assumed that the field is generated in individual cells of space in a statistically independent manner. A coupling between the cells is realized by magnetic diffusion. The rates of growth of the field and its moments are calculated. The field distribution generated by this mechanism possesses an intermittent nature in both time and space. PMID- 12779972 TI - Quantum chaos of atoms in a resonant cavity. AB - A system of atoms interacting with a radiation field in a resonant cavity is studied under conditions when the dynamics in the classical limit is stochastic. This situation is called quantum chaos. Equations of motion are obtained for the quantum-mechanical expectation values which take into account the quantum correlation functions. It is shown that in a situation corresponding to quantum chaos, the quantum corrections grow exponentially, making the evolution of the system essentially quantal after a certain time tau( variant Planck's over 2pi ) has elapsed. Analytical and numerical analysis show that in this regime the time tau( variant Planck's over 2pi ) obeys the logarithmic law tau( variant Planck's over 2pi ) approximately ln N (N is the number of atoms), and not the law tau( variant Planck's over 2pi ) approximately N(alpha) (alpha is a certain constant of order unity), as would be the case in the absence of chaos. PMID- 12779973 TI - Cantori for the stadium billiard. AB - Although almost all orbits in the stadium billiard are "chaotic," there are many regular orbits as well-the ordered periodic orbits and cantori. The symmetries of the stadium are exploited to find maximizing and saddle periodic orbits. Cantori are maximizing quasiperiodic orbits; they have caustics. Transport in the stadium should be impeded by cantori, just as in any twist map; this is particularly important for those orbits that are nearly glancing. PMID- 12779974 TI - Erratum: Varieties of spiral wave behavior: An experimentalist's approach to the theory of excitable media [CHAOS 1, 303-334 (1991)]. PMID- 12779975 TI - Overview of coupled map lattices. AB - Studies in coupled map lattices are briefly surveyed in connection with the papers in the present focus issue. PMID- 12779976 TI - Coupled trivial maps. AB - The first nontrivial example of coupled map lattices that admits a rigorous analysis in the whole range of the strength of space interactions is considered. This class is generated by one-dimensional maps with a globally attracting superstable periodic trajectory that are coupled by a diffusive nearest-neighbor interaction. PMID- 12779977 TI - Periodic orbits in coupled Henon maps: Lyapunov and multifractal analysis. AB - A powerful algorithm is implemented in a 1-d lattice of Henon maps to extract orbits which are periodic both in space and time. The method automatically yields a suitable symbolic encoding of the dynamics. The arrangement of periodic orbits allows us to elucidate the spatially chaotic structure of the invariant measure. A new family of specific Lyapunov exponents is defined, which estimate the growth rate of spatially inhomogeneous perturbations. The specific exponents are shown to be related to the comoving Lyapunov exponents. Finally, the zeta-function formalism is implemented to analyze the scaling structure of the invariant measure both in space and time. PMID- 12779978 TI - Space-time renormalization at the onset of spatio-temporal chaos in coupled maps. AB - The transition regime to spatio-temporal chaos via the quasiperiodic route as well as the period-doubling route is examined for coupled-map lattices. Space time renormalization-group analysis is carried out and the scaling exponents for the coherence length, the Lyapunov exponent, and the size of the phase fluctuations are determined. Universality classes for the different types of coupling at various routes to chaos are identified. PMID- 12779979 TI - Collective behaviors in coupled map lattices with local and nonlocal connections. AB - After having recalled the basic properties of the nontrivial collective dynamics exhibited by lattices of maps with local coupling and synchronous updating, we present the behavior of the same models in which all the connections are random. The mean-field, synchronized limit is shown to be reached only for large enough connectivities and sufficiently strong local chaos. Intermediate models, in which only a few of the connections of each site are taken at random, are then considered. Preliminary results indicate that the nontrivial collective behaviors shown by the regularly connected models may be robust to a small proportion of nonlocal, random connections. PMID- 12779980 TI - Describing the dynamics with a bi-orthogonal decomposition. AB - A space-time decomposition of a signal into orthogonal temporal modes (chronos) and orthogonal spatial modes (topos) for the analysis of the dynamics of CML closed to a Hopf bifurcation are used. Global energy and entropy as a tool to distinguish different kinds of dynamics in CML are introduced. PMID- 12779981 TI - Periodic orbits in a two-variable coupled map. AB - Periodic orbits are calculated for a linear transformation composed of two coupled tent maps using a symbolic dynamics defined as the direct product of the single-map symbols {0,1,2}. As the coupling strength is increased orbits are pruned and a crossover to one-dimensional behavior is observed. The disallowed binary orbits containing only symbols {0,1} form a connected region in a binary symbol plane. Stable orbits may appear for strong couplings. PMID- 12779982 TI - Coupled maps and pattern formation on the Sierpinski gasket. AB - The bifurcation structure of coupled maps on the Sierpinski gasket is investigated. The fractal character of the underlying lattice gives rise to stability boundaries for the periodic synchronized states with unusual features and spatially inhomogeneous states with a complex structure. The results are illustrated by calculations on coupled quadratic and cubic maps. For the coupled cubic map lattice bistability and domain growth processes are studied. PMID- 12779983 TI - Coupled map lattice techniques for simulating interfacial phenomena in reaction diffusion systems. AB - Interfacial patterns arise due to the dynamical evolution of phase boundaries in physical, chemical and biological systems. Coupled map lattices (CML) offer a useful tool for the simulation of such systems, being able to naturally accommodate the disparate length and time scales inherent in the dynamics of these processes. We illustrate this idea by reviewing work done on applying CML methods to crystal growth and to excitable media. PMID- 12779984 TI - Phenomenology of boiling: A coupled map lattice model. AB - A minimal model for boiling is proposed. With increasing temperature of a bottom plate, the model shows three successive phases; conduction, nucleate, and film boiling. In the nucleate regime the heat flux increases with the temperature of the bottom plate, while it decreases in the film boiling regime. In the boiling phase, the maximum Lyapunov exponent is positive, implying that the boiling phenomena are spatiotemporally chaotic. PMID- 12779985 TI - The plastic coupled map lattice: A novel image-processing paradigm. AB - Coupled map lattices (CML) can describe many relaxation and optimization algorithms currently used in image processing. We recently introduced the "plastic-CML" as a paradigm to extract (segment) objects in an image. Here, the image is applied by a set of forces to a metal sheet which is allowed to undergo plastic deformation parallel to the applied forces. In this paper we present an analysis of our "plastic-CML" in one and two dimensions, deriving the nature and stability of its stationary solutions. We also detail how to use the CML in image processing, how to set the system parameters and present examples of it at work. We conclude that the plastic-CML is able to segment images with large amounts of noise and large dynamic range of pixel values, and is suitable for a very large scale integration (VLSI) implementation. PMID- 12779986 TI - Coupled map lattices as computational systems. AB - The coupled map lattice (CML) as a mathematical model for a computer is considered. Using the theory of synchronous concurrent algorithms, it is shown that the CML is a valid new model for a parallel deterministic analog machine, but that, in principle, such a CML computer does not generate computations that cannot be reproduced by the standard mathematical models for computing on real numbers. The analysis is based on new general mathematical definitions of CMLs, and an axiomatic approach to determining which models of computation can be used to simulate CMLs. PMID- 12779987 TI - A neural network model as a globally coupled map and applications based on chaos. AB - First, a neural network model as the globally coupled map (GCM) is proposed. The model is obtained by modification of a Hopfield network model that has a negative self-feedback connection. Second, information processed by this model is interpreted in terms of the variety of the maps acting on the network elements, and a new, dynamic information processing model is described. The search for information using vague keywords, and solution of the traveling salesman problem (TSP) are introduced as applications. PMID- 12779988 TI - Nonequilibrium dynamics in lattice ecosystems: Chaotic stability and dissipative structures. AB - A generalized coupled map lattice (CML) model of ecosystem dynamics is presented. We consider the spatiotemporal behavior of a prey-predator map, a model of host parasitoid interactions, and two-species competition. The latter model can show phase separation of domains (Turing-like structures) even when chaos is present. We also use this CML model to explore the time evolution and structural properties of ecological networks built with a set of N competing species. The May-Wigner criterion is applied as a measure of stability, and some regularities in the stable networks observed are discussed. PMID- 12779989 TI - Evolution of host-parasitoid network through homeochaotic dynamics. AB - Host-parasitoid systems with evolving mutation rates are studied. By increasing the growth rate of hosts, the diversity of both species is maintained dynamically. For the lower growth rate, diversity is brought about by mere parasitism. The average mutation rate for parasites is elevated to a high value, while that for hosts is suppressed at a low level. For the higher growth rate, the mutation rates for both hosts and parasites are elevated to form a symbiotic cluster connected by on-going mutation. This symbiotic state is sustained through a chaotic oscillation keeping some coherency among species. For a flat landscape for hosts, dynamical clustering of oscillation is observed. Lyapunov spectra of such oscillations show that high dimensional chaos with small positive exponents underlies in the symbiotic state. This weak high dimensional chaos, termed "homeochaos," is essential to the maintenance of symbiosis in ecosystems. PMID- 12779990 TI - Semiclassical Poincare map for integrable systems. AB - The semiclassical Poincare map is applied to integrable systems and in particular to the rectangular billiard. The zeroes of the functional determinant are shown to give EBK quantization. The transfer operator is explicitly unitary and finite, resulting in a finite expansion of the Euler product over periodic orbits. PMID- 12779991 TI - Interpolating Hamiltonians for a stochastic-web map with quasicrystalline symmetry. AB - A systematic Hamiltonian approximation scheme is developed for a stochastic-web map with fivefold quasicrystalline symmetry. Interpolating Hamiltonians are calculated up to tenth order in the control parameter a. The higher order Hamiltonians are used to provide bounds for closed invariant curves of the map, and to investigate the structural evolution of map's phase portrait for a/=3/2), and for a suitably chosen classical particle with orbital angular momentum. A simple generalization of the PKQS model gives rise to stochastic webs on the surface of the unit sphere very similar to the Zaslavsky stochastic webs in a plane. PMID- 12780007 TI - Viscous attractor for the Galton board. AB - We analyze the Galton Board [or periodic "Lorentz Gas"] with a point mass scattered by elastic disks of diameter sigma, using a constant driving field g and a constant-viscosity linear drag force -p/tau, where p is the point-mass momentum. This combination leads to a nonequilibrium steady state which depends only upon the dimensionless ratio gtau(2)/sigma. The long-time-averaged trajectory leads to multifractal phase-space structures closely resembling those we found earlier using isokinetic equations of motion derived from Gauss' Principle of Least Constraint. A highly damped [small tau] creeping-flow limit describes our results for gtau(2)/sigma less than about 0.2. The lightly damped Green-Kubo linear-response limit for the model provides an accurate description of the dissipative dynamics for gtau(2)/sigma greater than about 2.0. PMID- 12780008 TI - Symmetric patterns in linear arrays of coupled cells. AB - In this note we show how to find patterned solutions in linear arrays of coupled cells. The solutions are found by embedding the system in a circular array with twice the number of cells. The individual cells have a unique steady state, so that the patterned solutions represent a discrete analog of Turing structures in continuous media. We then use the symmetry of the circular array (and bifurcation from an invariant equilibrium) to identify symmetric solutions of the circular array that restrict to solutions of the original linear array. We apply these abstract results to a system of coupled Brusselators to prove that patterned solutions exist. In addition, we show, in certain instances, that these patterned solutions can be found by numerical integration and hence are presumably asymptotically stable. PMID- 12780009 TI - Molecular Turing structures in the biochemistry of the cell. AB - Reactive lattice gas automata simulations show that Turing structure can form on a mesoscopic scale and are stable to molecular fluctuations in this domain. Calculations on the Sel'kov model suggest that Turing instabilities can give rise to global spatial symmetry breaking in ATP concentration within the cell cytoplasm with a mesoscopic Turing scale well within typical cell dimensions. This leads to a new mechanism for the global breaking of energy distribution in the cell. It also leads to reappraisal of the importance of the Turing effect on extended biochemical spatial structures and energy transport available to cell morphogenesis. PMID- 12780010 TI - Complexity in spiral wave dynamics(a)). AB - In closed systems of the Belousov-Zhabotinsky reaction a large number of dynamic states found in open systems is sampled as they evolve in time. During such slow aging processes of thin solution layers, prepared under appropriately chosen chemical conditions, an unexpectedly rich variety of spiral tip behavior was observed experimentally. Within a (concentration, time) parameter plane, the movement of free ends of waves was classified as follows: (a) in a stable domain periodic rigid rotation with cores of small (200 &mgr;m) or very large (2 mm) diameter; quasiperiodic compound motion along a hypocycle, a straight loopy line or an epicycle; complex meandering composed of possibly more than two components; (b) rectilinear tip motion indicating the boundary of spiral wave stability; and (c) in an unstable domain-shrinking of open ends of wave fronts during propagation. The main properties of these parameters are compared with recently published computer calculations. PMID- 12780011 TI - Turbulent decomposition of chemical waves by spontaneously induced hydrodynamic oscillation. AB - The dynamic evolution of a chemical reaction-diffusion pattern and its interaction with hydrodynamic flow is investigated by two-dimensional velocimetry and spectrophotometry based on microscope video imaging techniques. Oscillatory deformation and turbulent decomposition of chemical wave fronts are observed which are induced by a pronounced oscillatory flow excited spontaneously in a Belousov-Zhabotinsky solution layer with a free surface. PMID- 12780012 TI - Rescaled range analysis of resting respiration. AB - Fluctuations in resting depth of breathing (tidal volume) at constant breathing rate in the anesthetized adult rat exhibit fractal properties when analyzed by a rescaled range method characterized by a mean (+/-SD) exponent H=0.83+/-0.02 and 0.92+/-0.03 with and without sighs, respectively, for up to 400 breaths. Values of H determined from shuffled tidal volumes and simulated tidal volumes taken randomly from a Gaussian distribution of mean and variance approximating that of the actual data are consistent with the expected value of H=0.5 for an independent random process with finite variances. An empirical description is proposed to predict the change in H with length of time record. PMID- 12780013 TI - Statistical properties of chaos demonstrated in a class of one-dimensional maps. AB - One-dimensional maps with complete grammar are investigated in both permanent and transient chaotic cases. The discussion focuses on statistical characteristics such as Lyapunov exponent, generalized entropies and dimensions, free energies, and their finite size corrections. Our approach is based on the eigenvalue problem of generalized Frobenius-Perron operators, which are treated numerically as well as by perturbative and other analytical methods. The examples include the universal chaos function relevant near the period doubling threshold. Special emphasis is put on the entropies and their decay rates because of their invariance under the most general class of coordinate changes. Phase-transition like phenomena at the border state of chaos due to intermittency and super instability are presented. PMID- 12780014 TI - Periodically kicked hard oscillators. AB - A model of a hard oscillator with analytic solution is presented. Its behavior under periodic kicking, for which a closed form stroboscopic map can be obtained, is studied. It is shown that the general structure of such an oscillator includes four distinct regions; the outer two regions correspond to very small or very large amplitude of the external force and match the corresponding regions in soft oscillators (invertible degree one and degree zero circle maps, respectively). There are two new regions for intermediate amplitude of the forcing. Region 3 corresponds to moderate high forcing, and is intrinsic to hard oscillators; it is characterized by discontinuous circle maps with a flat segment. Region 2 (low moderate forcing) has a certain resemblance to a similar region in soft oscillators (noninvertible degree one circle maps); however, the limit set of the dynamics in this region is not a circle, but a branched manifold, obtained as the tangent union of a circle and an interval; the topological structure of this object is generated by the finite size of the repelling set, and is therefore also intrinsic to hard oscillators. PMID- 12780015 TI - Observing a codimension-two heteroclinic bifurcation. AB - This paper reports experimental observations of codimension-two heteroclinic bifurcations in an autonomous third-order electrical circuit. The paper also reports confirmations by computer simulations. In the laboratory experiments, a pair of programmable resistors are used in order to adjust two bifurcation parameters. In the associated two-parameter space, several codimension-one bifurcation sets are experimentally measured to capture codimension-two bifurcation structures. All of these bifurcation sets are numerically confirmed by exact bifurcation equations which are derived from piecewise-linear circuit dynamics. PMID- 12780016 TI - Phase-locking for maps of a torus: a computer assisted study. AB - In the description of bifurcations in a family of maps of an n-torus it is natural to consider phase-locked regions in the parameter space that correspond approximately to the sets of parameter values for which the maps have invariant tori. The extreme case of phase-locking is resonance, where the torus map has a periodic orbit. We study a family of maps of an n-torus that only differ from a family of torus translations by a small nonlinear perturbation. The widths of the phase-locked regions for this family generally increase linearly with the perturbation amplitude. However, this growth varies to a higher power law for families of maps that are given by trigonometric polynomials (the so-called Mathieu-type maps). The exponent of the asymptotic power law can be found by simple arithmetic calculations that relate the spectrum of the trigonometric polynomial to the unperturbed translation. Perturbation theory and these calculations predict that typical resonance regions for the family of Mathieu type maps are narrow elliptical annuli. All these results are illustrated in a number of numerical examples. PMID- 12780017 TI - Torus knots and polynomial invariants for a class of soliton equations. AB - In this paper is shown how to interpret the nonlinear dynamics of a class of one dimensional physical systems exhibiting soliton behavior in terms of Killing fields for the associated dynamical laws acting as generators of torus knots. Soliton equations are related to dynamical laws associated with the intrinsic kinematics of space curves and torus knots are obtained as traveling wave solutions to the soliton equations. For the sake of illustration a full calculation is carried out by considering the Killing field that is associated with the nonlinear Schrodinger equation. Torus knot solutions are obtained explicitly in cylindrical polar coordinates via perturbation techniques from the circular solution. Using the Hasimoto map, the soliton conserved quantities are interpreted in terms of global geometric quantities and it is shown how to express these quantities as polynomial invariants for torus knots. The techniques here employed are of general interest and lead us to make some conjectures on natural links between the nonlinear dynamics of one-dimensional extended objects and the topological classification of knots. PMID- 12780018 TI - 3-D spatial chaos in the elastica and the spinning top: Kirchhoff analogy. AB - The existence of spatially chaotic deformations in an elastica and the analogous motions of a free spinning rigid body, an extension of the problem originally examined by Kirchhoff are investigated. It is shown that a spatially periodic variation in cross sectional area of the elastica results in spatially complex deformation patterns. The governing equations for the elastica were numerically integrated and Poincare maps were created for a number of different initial conditions. In addition, three dimensional computer images of the twisted elastica were generated to illustrate periodic, quasiperiodic, and stochastic deformation patterns in space. These pictures clearly show the existence of spatially chaotic deformations with stunning complexity. This finding is relevant to a wide variety of fields in which coiled structures are important, from the modeling of DNA chains to video and audio tape dynamics to the design of deployable space structures. PMID- 12780019 TI - Ray dynamics of the propagation of sound in nonuniform moving media. AB - This paper presents a new ray theory for the propagation of sound waves in nonuniformly moving media. It is found that the ray equations in weakly inhomogeneous and slowly moving media are analogous to the equations of motion of charged particles in nonuniform electric and magnetic fields. The adiabatic approximation is used to study the problem of the propagation of sound rays in a model of near-ocean-bottom waveguide with horizontal flow and slowly varying parameters along the direction of propagation of the wave. A general formula is derived that describes the transverse displacement of the trajectory of the ray relative to the direction of propagation of the wave. PMID- 12780020 TI - Waveguide propagation of intense electromagnetic radiation in slightly inhomogeneous nonlinear media. AB - The propagation of self-localizing beams of electromagnetic waves in the form of nonlinear waveguides in a slightly inhomogeneous medium is studied analytically and numerically. The trajectories of the axial ray are studied as a function of its direction and the field strength at the initial point on the basis of a nonlinear scalar Helmholtz equation. Analytic expressions are derived. The longitudinal refractive index, the field intensity, and the waveguide radius are plotted as functions of the instantaneous position of the point on the axial ray. Deep penetration of the beam into the opaque region and the position of the screening surface are studied as functions of the parameters of the beam and the medium. A steady-state 3D problem is analyzed for a power-law nonlinearity with an arbitrary power. A 2D problem is analyzed for the case of a ponderomotive nonlinearity with saturation. PMID- 12780021 TI - On the equality of Hausdorff and box counting dimensions. AB - By viewing the covers of a fractal as a statistical mechanical system, the exact capacity of a multifractal is computed. The procedure can be extended to any multifractal described by a scaling function to show why the capacity and Hausdorff dimension are expected to be equal. PMID- 12780022 TI - On noise reduction methods for chaotic data. AB - Recently proposed noise reduction methods for nonlinear chaotic time sequences with additive noise are analyzed and generalized. All these methods have in common that they work iteratively, and that in each step of the iteration the noise is suppressed by requiring locally linear relations among the delay coordinates, i.e., by moving the delay vectors towards some smooth manifold. The different methods can be compared unambiguously in the case of strictly hyperbolic systems corrupted by measurement noise of infinitesimally low level. It was found that all proposed methods converge in this ideal case, but not equally fast. Different problems arise if the system is not hyperbolic, and at higher noise levels. A new scheme which seems to avoid most of these problems is proposed and tested, and seems to give the best noise reduction so far. Moreover, large improvements are possible within the new scheme and the previous schemes if their parameters are not kept fixed during the iteration, and if corrections are included which take into account the curvature of the attracting manifold. Finally, the fact that comparison with simple low-pass filters tends to overestimate the relative achievements of these nonlinear noise reduction schemes is stressed, and it is suggested that they should be compared to Wiener-type filters. PMID- 12780023 TI - Measuring the similarity between trajectories using clustering techniques. AB - A clustering method has been developed to group signals that display similar dynamic behavior. The procedure involves using the method of time delay embedding to construct a trajectory in state space from a time series. Certain features that characterize the geometry of the trajectory have been defined. These features were subjected to a series of statistical tests to determine their usefulness in a hierarchical clustering analysis. The latter is aimed at finding groups of similar trajectories. The trajectory-based clustering algorithm has been applied to simulated data, which included both stochastic data generated by a linear AR model, and nonlinear data generated by a Duffing oscillator. The results show that the algorithm works reliably in both cases. PMID- 12780024 TI - Exit times and transport for symplectic twist maps. AB - The exit time decomposition of a set yields a description of the transport through the set as well as a visualization of the invariant structures inside it. We construct several sets computationally easier to deal with than the construction of resonances, based on the ordering properties for orbits of twist maps. Furthermore these sets can be constructed for four- and higher-dimensional twist mappings. For the four-dimensional case-using the example of Froeshle-we find "practically" invariant volumes surrounding elliptic fixed points. The boundaries of these regions are remarkably sharp; however, the regions are threaded by "tubes" of escaping orbits. PMID- 12780025 TI - Solution multistability in first-order nonlinear differential delay equations. AB - The dependence of solution behavior to perturbations of the initial function (IF) in a class of nonlinear differential delay equations (DDEs) is investigated. The structure of basins of attraction of multistable limit cycles is investigated. These basins can possess complex structure at all scales measurable numerically although this is not necessarily the case. Sensitive dependence of the asymptotic solution to perturbations in the initial function is also observed experimentally using a task specific electronic analog computer designed to investigate the dynamics of an integrable first-order DDE. PMID- 12780026 TI - A method of investigation of the evolution of a nonlinear system driven by pulsed noise. AB - A numerical method is proposed for determining the evolution of nonlinear systems subjected to noise. The method is based on a recurrence equation for the probability density which has been obtained analytically due to the choice of noise in the form of discrete series of random pulses. The method is applied to a dynamical system which describes the motion of a particle in a plane-wave field. The evolution of the probability density in phase and energy space is obtained. It is shown that because of noise effects, the region in phase space where particles can be found rapidly reaches the separatrix and then spreads over the phase space, mainly along the separatrix. In the energy spectrum a new peak appears at the separatrix's energy. This peak grows in time, while the main peak corresponding to the initial energy drops in time and shifts to lower energy. The moments of motion were analyzed. The character of their evolution indicates a high rate of chaotization. The growth of the fraction of energetic particles is very rapid (exponential at the beginning), whereas the mean energy grows linearly. PMID- 12780027 TI - Chaos in the one-dimensional gravitational three-body problem. AB - We have investigated the appearance of chaos in the one-dimensional Newtonian gravitational three-body system (three masses on a line with -1/r pairwise potential). In the center of mass coordinates this system has two degrees of freedom and can be conveniently studied using Poincare sections. We have concentrated in particular on how the behavior changes when the relative masses of the three bodies change. We consider only the physically more interesting case of negative total energy. For two mass choices we have calculated 18 000 full orbits (with initial states on a 100x180 lattice on the Poincare section) and obtained dwell time distributions. For 105 mass choices we have calculated Poincare maps for 10x18 starting points. Our results show that the Poincare section (and hence the phase space) divides into three well defined regions with orbits of different characteristics: (1) There is a region of fast scattering, with a minimum of pairwise collisions. This region consists of 'scallops' bordering the E=0 line, within a scallop the orbits vary smoothly. The number of the scallops increases as the mass of the central particle decreases. (2) In the chaotic scattering region the interaction times are longer, and both the interaction time and the final state depend sensitively on the starting point on the Poincare section. For both (1) and (2) the initial and final states consist of a binary + single particle. (3) The third region consists of quasiperiodic orbits where the three masses are bound together forever. At the center of the quasiperiodic region there is a periodic orbit discovered (numerically) by Schubart in 1956. The stability of the Schubart orbit turns out to correlate strongly with the global behavior. PMID- 12780028 TI - Deterministic chaos in a quantum mechanical system described by a time independent Hamiltonian: A model of three Josephson junctions in a loop. AB - Quantum mechanical equations of motion are obtained for a system consisting of a very large number of three types of interacting bosons. Under suitable choice of parameters of the Hamiltonian of the system, the equations of motion are those describing three Josephson junctions in a superconducting loop. It is shown numerically that this system is capable of exhibiting deterministic chaos (extreme sensitivity to initial conditions). PMID- 12780029 TI - New cases of quasiperiodic motions in reversible systems. AB - A theorem on the existence of invariant D-dimensional tori in reversible mappings near surfaces foliated into invariant tori of dimension d is announced, where d union or logical sum (i=0) (n 1)S(1) (i) which maps each circle onto another by a rotation. This particular type of interval exchange map arises naturally in bifurcation theory. In this paper we give a full description of the symbolic dynamics associated to such maps. PMID- 12780117 TI - kltool: A tool to analyze spatiotemporal complexity. AB - We announce the availability of a software package, called kltool, that can extract phase space information from complex spatiotemporal data via the Karhunen Loeve analysis. Data generated by the periodic, quasiperiodic or chaotic evolution of a small number of spatially coherent structures can be processed. A key feature of kltool is that it allows the user to interact easily with the data processing and its graphical display. We illustrate the use of kltool on numerical data from the Kuramoto-Sivashinsky equation and laboratory data from a flame experiment. PMID- 12780118 TI - From oscillations to excitability: A case study in spatially extended systems. AB - This volume is devoted to the presentation of the main contributions to the workshop "From oscillations to excitability: A case study in spatially extended systems," organized by the authors in Nice in June 1993. It gives an overview of the current research on spatiotemporal patterns in a wide range of systems that display self-oscillatory or excitable behavior. It tries to give a better understanding of the transition from the oscillatory to the excitable regime and of its effect on the properties of spiral waves, and to fill the gap between the theories and concepts used to describe both regimes in the so-called "active media." PMID- 12780119 TI - The Ginzburg-Landau approach to oscillatory media. AB - Close to a supercritical Hopf bifurcation, oscillatory media may be described, by the complex Ginzburg-Landau equation. The most important spatiotemporal behaviors associated with this dynamics are reviewed here. It is shown, on a few concrete examples, how real chemical oscillators may be described by this equation, and how its coefficients may be obtained from the experimental data. Furthermore, the effect of natural forcings, induced by the experimental realization of chemical oscillators in batch reactors, may also be studied in the framework of complex Ginzburg-Landau equations and its associated phase dynamics. We show, in particular, how such forcings may locally transform oscillatory media into excitable ones and trigger the formation of complex spatiotemporal patterns. PMID- 12780120 TI - A dynamical systems approach to spiral wave dynamics. AB - A simple system of five nonlinear ordinary differential equations is shown to reproduce many dynamical features of spiral waves in two-dimensional excitable media. PMID- 12780121 TI - Electrical alternans and spiral wave breakup in cardiac tissue. AB - This paper reports the results of a theoretical investigation of spiral wave breakup in model equations of action potential propagation in cardiac tissue. A general formulation of these equations is described in which arbitrary experimentally determined restitution and dispersion curves can in principle be fitted. Spiral wave behavior is studied in two-dimension as a function of a parameter Re which controls the steepness of the restitution curve at short diastolic intervals. Spiral breakup is found to occur when the minimum period T(min), below which a periodically stimulated tissue exhibits alternans in action potential duration, exceeds by a finite amount the spiral rotation period T(S). At this point, oscillations in action potential duration are of sufficiently large amplitude to cause a spontaneous conduction block to form along the wavefront. The latter occurs closer to the initiation point of reentry (spiral tip) with increasing steepness and, hence, in smaller tissue sizes. Spiral breakup leads to a spatially disorganized wave activity which is always transient, except for tissues larger than some minimum size and within a very narrow range of Re which increases with dispersion. PMID- 12780122 TI - Mechanisms of target and spiral wave propagation in single cells. AB - Target and spiral wave propagation have been observed in single cells such as myocites. Moreover, in the same cells, transition from target waves to planar waves or from the latter to spiral waves was also observed. Considering an oscillatory medium described by the Ginzburg-Landau equation we suggest that such phenomena could be explained if cell nuclei and cell organelles are considered as obstacles in a small bounded medium. We discuss the role of cell geometry as well as the phenomenon of reentry at the cellular level. PMID- 12780123 TI - Complex patterns in reaction-diffusion systems: A tale of two front instabilities. AB - Two front instabilities in a reaction-diffusion system are shown to lead to the formation of complex patterns. The first is an instability to transverse modulations that drives the formation of labyrinthine patterns. The second is a nonequilibrium Ising-Bloch (NIB) bifurcation that renders a stationary planar front unstable and gives rise to a pair of counterpropagating fronts. Near the NIB bifurcation the relation of the front velocity to curvature is highly nonlinear and transitions between counterpropagating fronts become feasible. Nonuniformly curved fronts may undergo local front transitions that nucleate spiral-vortex pairs. These nucleation events provide the ingredient needed to initiate spot splitting and spiral turbulence. Similar spatiotemporal processes have been observed recently in the ferrocyanide-iodate-sulfite reaction. PMID- 12780124 TI - Excitability in liquid crystal. AB - The spiral waves observed in a liquid crystal submitted to a vertical electric field and a horizontal rotating magnetic field are explained in the framework of a purely mechanical description of the liquid crystal. The originality of the experiment described in this paper is the presence of the vertical electric field which allows us to analyze the spiral waves in the framework of a weakly nonlinear theory. PMID- 12780125 TI - Pattern formation and competition in photorefractive oscillators. AB - We introduce a general model of pattern formation in optical systems made of a cavity with an active medium as a photorefractive crystal fed by a pump. The model is based on the interplay of a diffractive equation for the optical field and a diffusive equation for the medium refractivity. The aim of the model is to describe a series of experiments which have shown mode competition (periodic or chaotic alternation) for low Fresnel numbers (F) and mode coexistence, leading to short range space correlations, for high F. For low F, a linear stability analysis provides the set of modes above threshold as a function of the transverse wave number. Due to the interplay of the optical and the diffusive interactions, different behaviors result depending on the thickness of the medium as compared to the optical absorption length and diffusion length. Including the leading nonlinearities compatible with the symmetry constraints, we introduce normal form equations which describe the time-dependent mode competition. In the case of a large number of modes (high F), nonlinear mode-mode interaction is equivalent to a self-induced noise. In this limit, the relevant feature to be compared with the experiment is the power spectrum. PMID- 12780126 TI - Chemical turbulence and standing waves in a surface reaction model: The influence of global coupling and wave instabilities. AB - Among heterogeneously catalyzed chemical reactions, the CO oxidation on the Pt(110) surface under vacuum conditions offers probably the greatest wealth of spontaneous formation of spatial patterns. Spirals, fronts, and solitary pulses were detected at low surface temperatures (T<500 K), in line with the standard phenomenology of bistable, excitable, and oscillatory reaction-diffusion systems. At high temperatures (T greater, similar 540 K), more surprising features like chemical turbulence and standing waves appeared in the experiments. Herein, we study a realistic reaction-diffusion model of this system, with respect to the latter phenomena. In particular, we deal both with the influence of global coupling through the gas phase on the oscillatory reaction and the possibility of wave instabilities under excitable conditions. Gas-phase coupling is shown to either synchronize the oscillations or to yield turbulence and standing structures. The latter findings are closely related to clustering in networks of coupled oscillators and indicate a dominance of the global gas-phase coupling over local coupling via surface diffusion. In the excitable regime wave instabilities in one and two dimensions have been discovered. In one dimension, pulses become unstable due to a vanishing of the refractory zone. In two dimensions, turbulence can also emerge due to spiral breakup, which results from a violation of the dispersion relation. PMID- 12780127 TI - External forcing of spiral waves. AB - The effect of an external rhythm on rotating spiral waves in excitable media is investigated. Parameters of the unperturbed medium were chosen, such that the organizing spiral tip describes meandering (hypocyclic) trajectories, which are the most general shape for the experimentally observed systems. Periodical modulation of excitability in a model of the Belousov-Zhabotinsky (BZ) reaction forces meandering spiral tips to describe trajectories that are not found at corresponding stationary conditions. For different modulation periods, two types of resonance drift, phase-locked tip motion, a spectrum of hypocyclic trajectories, and complex multifrequency patterns were computed. The computational results are complemented by experimental data obtained for periodically changing illumination of the photosensitive BZ reaction. The observed drastic deformation of the tip trajectory is considered as an efficient means to study and to control wave processes in excitable media. PMID- 12780128 TI - Spiral breakup induced by an electric current in a Belousov-Zhabotinsky medium. AB - Sprial breakup in the Belousov-Zhabotinsky reaction has been observed under the influence of an externally applied alternating electric current. The dynamic mechanism of this breakup is explained in the framework of this reaction. The dependence of the critical electric current amplitude on the period of the wave and on the excitability of the medium is analyzed. Spiral breakup is shown to provide a limit of validity of electric-field-induced drift of vortices in excitable media. Experimental results are complemented with numerical simulations provided by two- and three-variable Oregonator models. PMID- 12780129 TI - High-frequency instability of wave fronts. AB - Experimental evidence is presented that a lateral instability of a wave front, as described earlier in a chemically active medium with the Belousov-Zhabotinsky reaction with decreased excitability, can also occur in a medium with any degree of excitability provided that a high-frequency wave train travels through the medium. The interaction of chemical waves with the boundary of the medium can result in the appearance of wave breaks and spiral waves. PMID- 12780130 TI - Spatiotemporal chaos in an electric current driven ionic reaction-diffusion system. AB - Two types of transitions from the time-periodic spatiotemporal patterns to chaotic ones in the spatially one-dimensional ionic reaction-diffusion system forced either with direct or alternating electric field are described and analyzed by numerical techniques. An ionic version of the Brusselator kinetic scheme is considered. The Karhunen-Loeve decomposition technique is shown to be a possible tool for the global representation of dynamic behavior, but fails as a tool in the identification of the route of transition to chaos in the case of direct current forcing. Higher dimensional chaos with two positive Lyapunov exponents has been identified for the case of alternating current forcing. Results of the Karhunen-Loeve analysis are compared to results of classical analysis of local time series (attractor dimensions, Lyapunov exponents). PMID- 12780131 TI - Effect of an externally applied electric field on excitation propagation in the cardiac muscle. AB - Classical theory of potential distribution in cardiac muscle (cable theory) postulates that all effects of electric field (internally or externally applied) should decay exponentially with a space constant of the order of the tissue space constant ( approximately 1 mm). Classical theory does not take into account the cellular structure of the heart. Here, we formulate a mathematical model of excitation propagation taking into account cellular gap junctions. Investigation of the model has shown that the classical description is correct on the macroscopic scale only. At microscopic scale, electric field is modulated with a spatial period equal to the cell size (Plonsey and Barr), with the zero average. A very important new feature found here is that this effect of electric field does not decay at arbitrary big distances from the electrode. It opens the new way to control the excitation propagation in the cardiac muscle. In particular, we show that electric field can modify the velocity of propagation of an impulse in cardiac tissue at arbitrary big distances from electrode. In 2-dimensions, it can make rotating waves drift. To test these predictions, experiments with cardiac preparations are proposed. PMID- 12780132 TI - Modeling excitable media by a one variable cellular automaton: Application to the cardiac case. AB - The dynamics of an assembly of cardiac cells is modeled by a simple cellular automaton that reduces to a single variable the two variable competition of the standard models of excitable media. Furthermore, a short superexcitability period is introduced, as suggested by the dynamics of the single cardiac miocyte. The model reproduces several pathological cardiac behaviors as, e.g., the fast transition from normal behavior to fibrillation, showing how this latter one can either occur over the whole spatial domain or can be confined within a limited region. PMID- 12780133 TI - Modeling spatial patterns in Dictyostelium. AB - The life cycle of Dictyostelium discoideum provides a striking example of the transition from single cell behavior to multicellular cooperativity. In this paper the status of the attempts at making semiquantitative models of the aggregation phase of this cycle is reviewed. Specifically, it is discussed how the propagation of cAMP waves is a typical example of excitable signaling, which is then rendered unstable by coupling to cell chemotaxis. To investigate the streaming pattern that emerges from this clumping instability, we next turn to a new simulation strategy, which couples dynamical cell-like entities ("bions") to continuum chemical concentration fields. Finally, we discuss two directions for further research: One is the study of the robustness with respect to the variation of system parameters (such as the cell density) exhibited by the biological system, but not by any simple model. The other concerns going beyond the aggregation phase to tackle the three-dimensional problem of slug formation and motion. PMID- 12780134 TI - Two-dimensional model of a kicked oscillator: Motion with intermittency. AB - The phenomenon of intermittency, which arises near a point of degeneracy of an unperturbed Hamiltonian under the influence of a discontinuous perturbation function, is studied in the example of a two-dimensional (2-D) model of a kicked oscillator. This example describes the dynamics of a particle in a cylindrically symmetric potential well subjected to radial kicks which occur periodically in time. The problem is reduced to a Hamiltonian system with N=3/2 degrees of freedom, whose unperturbed Hamiltonian has a degeneracy point. The intermittency is studied numerically and analytically. PMID- 12780135 TI - One-dimensional mappings of the eigenelements of the Schrodinger problem. AB - An approach is proposed for analyzing the inverse spectral problems for the Schrodinger equation based on writing the equation for the analog of the number of-quanta operator for a harmonic oscillator. This equation makes it possible to determine not only the one-dimensional mapping of the energy eigenvalues but also the linear equation for the point spectrum shift operator of the Schrodinger problem. The solvability conditions of the latter lead to a nonlinear equation that determines the class of allowable potentials. Two classes of potentials regular in R(1) and symmetrical are isolated on the basis of the proposed approach. The first of these leads to equidistant spectra with a gap of arbitrary size and location. The spectrum of the second potential class is a combination of three rigorously equidistant spectra with ground states that are shifted by an arbitrary amount. Generalizations to the case of essentially nonequidistant spectra are shown to be possible. PMID- 12780136 TI - The width of the exponentially narrow stochastic layers. AB - Exponentially small splitting of the separatrix has been calculated for a high frequency large amplitude perturbation and the correspondent correction to the width of the stochastic layer is obtained. The result can be applied to the large amplitude perturbation. PMID- 12780137 TI - Separatrices splitting for Birkhoff's billiard in symmetric convex domain, closed to an ellipse. AB - An example of a convex domain on the plane with the phenomenon of the transversal intersection of separatrices of the corresponding billiard mapping is presented. This example is constructed as an analytic global symmetric perturbation of an ellipse and we investigate the global symmetric analytic perturbation of the integrable billiard mapping in the ellipse. We establish a theorem on the separatrices splitting of the perturbed billiard mapping and derive the asymptotic formulas for a homoclinic invariant as well as for a "principal" splitting angle of separatrices, arising from the hyperbolic fixed point of the mapping. (c) 1994 American Institute of Physics. PMID- 12780138 TI - Chaotic domains: A numerical investigation. AB - We study the chaotic domain state in rotating convection using a model equation that allows for a continuous range of roll orientations as in the experimental system. Methods are developed for extracting the domain configuration from the resulting patterns that should be applicable to a wide range of domain states. Comparison with the truncated three mode amplitude equation description is made. (c) 1994 American Institute of Physics. PMID- 12780139 TI - Stability and bifurcations of a stationary state for an impact oscillator. AB - The motion of a vibroimpacting one-degree-of-freedom model is analyzed. This model is motivated by the behavior of a shearing granular material, in which a transitional phenomenon is observed as the concentration of the grains decreases. This transition changes the motion of a granular assembly from an orderly shearing between two blocks sandwiching a single layer of grains to a chaotic shear flow of the whole granular mass. The model consists of a mass-spring dashpot assembly that bounces between two rigid walls. The walls are prescribed to move harmonically in opposite phases. For low wall frequencies or small amplitudes, the motion of the mass is damped out, and it approaches a stationary state with zero velocity and displacement. In this paper, the stability of such a state and the transition into chaos are analyzed. It is shown that the state is always changed into a saddle point after a bifurcation. For some parameter combinations, horseshoe-like structures can be observed in the Poincare sections. Analyzing the stable and unstable manifolds of the saddle point, transversal homoclinic points are found to exist for some of these parameter combinations. (c) 1994 American Institute of Physics. PMID- 12780140 TI - Complexity of precipitation patterns: Comparison of simulation with experiment. AB - Numerical simulations show that a simple model for the formation of Liesegang precipitation patterns, which takes into account the dependence of nucleation and particle growth kinetics on supersaturation, can explain not only simple patterns like parallel bands in a test tube or concentric rings in a petri dish, but also more complex structural features, such as dislocations, helices, "Saturn rings," or patterns formed in the case of equal initial concentrations of the source substances. The limits of application of the model are discussed. (c) 1994 American Institute of Physics. PMID- 12780141 TI - Four modes competition and chaos in a shell. AB - Steady-state chaotic vibrations of a shallow shell as a system with a nonsymmetrical restoring force and one equilibrium state are considered. Mode interaction and its effect on a chaotic behavior of the shell is studied. The terms "natural" and "imposed" chaos are introduced for the response of resonant and nonresonant modes. It is shown that such a qualitative difference is important for better understanding of chaos in systems with distributed parameters, and may be very useful for numerical investigations. Some qualitative comparisons with previous papers on chaos in distributed mechanical systems are also made. (c) 1994 American Institute of Physics. PMID- 12780142 TI - Normally attracting manifolds and periodic behavior in one-dimensional and two dimensional coupled map lattices. AB - We consider diffusively coupled logistic maps in one- and two-dimensional lattices. We investigate periodic behaviors as the coupling parameter varies, i.e., existence and bifurcations of some periodic orbits with the largest domain of attraction. Similarity and differences between the two lattices are shown. For small coupling the periodic behavior appears to be characterized by a number of periodic orbits structured in such a way to give rise to distinct, reverse period doubling sequences. For intermediate values of the coupling a prominent role in the dynamics is played by the presence of normally attracting manifolds that contain periodic orbits. The dynamics on these manifolds is very weakly hyperbolic, which implies long transients. A detailed investigation allows the understanding of the mechanism of their formation. A complex bifurcation is found which causes an attracting manifold to become unstable. (c) 1994 American Institute of Physics. PMID- 12780143 TI - The kinetics of liquid-gas phase transitions of a Van der Waals substance with fluctuations taken into account. AB - The isotherm of a Van der Waals substance, containing only stable points, is obtained on the basis of using the thermodynamic potential for nonequilibrium states and taking fluctuations into account. It is shown that in the vicinity of two-phase states this isotherm is close to the horizontal phase equilibrium line, defined by Maxwell's rule. The lifetimes of the metastable states of the Van der Waals substance, which depend on the intensity of the external fluctuations and the number of particles in the system, are estimated. (c) 1994 American Institute of Physics. PMID- 12780144 TI - Regular and chaotic transport of impurities in steady flows. AB - This paper considers the properties of the transport of impurity particles in steady fluid flows and describes the principal modes of particle motion. An impurity consisting of particles with a lower density than that of the medium is localized at stationary points of the flow, whereas a heavy impurity can perform a spatially unbounded motion. The conditions for the transition from the bounded motion of a heavy impurity to the long-range transport mode, which occurs as a result of a loss of the stability of the heteroclinic trajectory, are obtained for a model two-dimensional flow having an eddy-cell structure. A mode is found in which a particle, after being transported over a long distance, is trapped forever within the confines of one cell. The transition from regular to chaotic particle transport is analyzed. The question of the effect of a small noise (for example, molecular diffusion) on the character of the motion of a heavy impurity is investigated. It is shown that this effect is important at high viscosity and leads to a transition from bounded motion of the impurity particle to diffusion type chaotic motion. (c) 1994 American Institute of Physics. PMID- 12780145 TI - Chaotic capture of vortices by a moving body. II. Bound pair model. AB - Previously, we have presented a simple model for the interaction of a fluid vortex structure with a moving bluff body, and demonstrated the existence of a trapping mechanism related to chaotic scattering. This single point vortex model required explicit perturbation to generate chaos and the subsequent complex dynamics. Here, we present a model which attempts to introduce internal degrees of-freedom in the vortex structure in the simplest manner, by replacing the single vortex with a like-signed pair. We show that this model exhibits chaotic trapping without the need of explicit perturbation, however, the region of parameter space for which trapping occurs is exceedingly small due to the spatially dependent form of the perturbation. We claim that this result explains some the behavior observed in Navier-Stokes simulations of the same vortex-body system, where we find close correspondence between the dynamics of an extended vorticity distribution and the single vortex model. Finally, we generalize the model to unequal strength vortex pairs, and find more complex behavior which includes "partial" capture of the weaker vortex by the body. (c) 1994 American Institute of Physics. PMID- 12780146 TI - Unsteady processes in machines. AB - Couplings in machines and mechanisms always have play and friction. While under loading, stick-slip phenomena and impact events can take place. Such processes are modeled as multibody systems whose structure is time variant or unsteady. The time-variant number of degrees of freedom is due to stick-slip contacts. The coupling characteristics become unsteady, for instance there exist jumps in the loads, if impacts occur. For establishing a uniform theory for such phenomena we use a Lagrangian approach connecting the additional constraint equations and the equations of motion by Lagrange multipliers, which are proportional to the constraint forces. Stick-slip and impact events are evaluated by indicator functions leading to special numerical algorithms for the search of switching points. Contact problems are formulated as a complementarity problem which can be solved by efficient algorithms. The theory is applied to rattling in gears, impact drilling machines, turbine blade dampers, and a woodpecker toy. In some of these applications, chaos as a result of bifurcations is possible, which results from variations in the parameters. (c) 1994 American Institute of Physics. PMID- 12780147 TI - Dynamical disease: Identification, temporal aspects and treatment strategies of human illness. AB - Dynamical diseases are characterized by sudden changes in the qualitative dynamics of physiological processes, leading to abnormal dynamics and disease. Thus, there is a natural matching between the mathematical field of nonlinear dynamics and medicine. This paper summarizes advances in the study of dynamical disease with emphasis on a NATO Advanced Research Worshop held in Mont Tremblant, Quebec, Canada in February 1994. We describe the international effort currently underway to identify dynamical diseases and to study these diseases from a perspective of nonlinear dynamics. Linear and nonlinear time series analysis combined with analysis of bifurcations in dynamics are being used to help understand mechanisms of pathological rhythms and offer the promise for better diagnostic and therapeutic techniques. (c) 1995 American Institute of Physics. PMID- 12780148 TI - Dynamic diseases in neurology and psychiatry. AB - Thirty-two (32) periodic diseases of the nervous system are identified in which symptoms and/or signs recur. In 10/32, the recurrence of a symptom complex is one of the defining features of the illness, whereas in 22/32 oscillatory signs occur in the setting of an ongoing nervous system disorder. We discuss the possibility that these disorders may be dynamic diseases. (c) 1995 American Institute of Physics. PMID- 12780149 TI - Hiccups as a dynamical disease. PMID- 12780150 TI - Dysrhythmias of the respiratory oscillator. AB - Breathing is regulated by a central neural oscillator that produces rhythmic output to the respiratory muscles. Pathological disturbances in rhythm (dysrhythmias) are observed in the breathing pattern of children and adults with neurological and cardiopulmonary diseases. The mechanisms responsible for genesis of respiratory dysrhythmias are poorly understood. The present studies take a novel approach to this problem. The basic postulate is that the rhythm of the respiratory oscillator can be altered by a variety of stimuli. When the oscillator recovers its rhythm after such perturbations, its phase may be reset relative to the original rhythm. The amount of phase resetting is dependent upon stimulus parameters and the level of respiratory drive. The long-range hypothesis is that respiratory dysrhythmias can be induced by stimuli that impinge upon or arise within the respiratory oscillator with certain combinations of strength and timing relative to the respiratory cycle. Animal studies were performed in anesthetized or decerebrate preparations. Neural respiratory rhythmicity is represented by phrenic nerve activity, allowing use of open-loop experimental conditions which avoid negative chemical feedback associated with changes in ventilation.In animal experiments, respiratory dysrhythmias can be induced by stimuli having specific combinations of strength and timing. Newborn animals readily exhibit spontaneous dysrhythmias which become more prominent at lower respiratory drives. In human subjects, swallowing was studied as a physiological perturbation of respiratory rhythm, causing a pattern of phase resetting that is characterized topologically as type 0. Computational studies of the Bonhoeffer van der Pol (BvP) equations, whose qualitative behavior is representative of many excitable systems, supports a unified interpretation of these experimental findings. Rhythmicity is observed when the BvP model exhibits recurrent periods of excitation alternating with refractory periods. The same system can be perturbed to a state in which amplitude of oscillation is attenuated or abolished. We have characterized critical perturbations which induce transitions between these two states, giving rise to patterns of dysrhythmic activity that are similar to those seen in the experiments. We illustrate the importance of noise in initiation and termination of rhythm, comparable to normal respiratory rhythm intermixed with spontaneous dysrhythmias. In the BvP system the incidence and duration of dysrhythmia is shown to be strongly influenced by the level of noise. These studies should lead to greater understanding of rhythmicity and integrative responses of the respiratory control system, and provide insight into disturbances in control mechanisms that cause apnea and aspiration in clinical disease states. (c) 1995 American Institute of Physics. PMID- 12780151 TI - Nonlinear dynamics of the voice: Signal analysis and biomechanical modeling. AB - Irregularities in voiced speech are often observed as a consequence of vocal fold lesions, paralyses, and other pathological conditions. Many of these instabilities are related to the intrinsic nonlinearities in the vibrations of the vocal folds. In this paper, bifurcations in voice signals are analyzed using narrow-band spectrograms. We study sustained phonation of patients with laryngeal paralysis and data from an excised larynx experiment. These spectrograms are compared with computer simulations of an asymmetric 2-mass model of the vocal folds. (c) 1995 American Institute of Physics. PMID- 12780152 TI - Tremor: Is Parkinson's disease a dynamical disease? AB - Experimental evidence has shown a plethora of short-term fluctuations in patients with Parkinson's disease. We investigate these transitory events using the concept of dynamical disease. Several examples of short-term fluctuations in tremor are analyzed, and in two cases, other systemic variables (i.e., respiration and blood pressure) are examined as well. A model for tremor, based on negative feedback with delays is proposed, and the transient events are simulated. The theoretical implications of the model suggest that interactions between the central and peripheral loops, as well as interactions between the control loops and other systemic signals, can give rise to transitory events in tremor, both in the pathological and in the normal case. (c) 1995 American Institute of Physics. PMID- 12780153 TI - The dynamical structure of tremor in tardive dyskinesia. AB - The movement disorder syndrome of tardive dyskinesia arises as a consequence of prolonged regimens of neuroleptic medication, and is characterized, although not exclusively, by jerky and sometimes rhythmical stereotypical motions in a wide range of muscle systems. It is well established that the degree and variability of tremor in tardive dyskinesia is greater than that in normal age-matched subjects. The findings from the current experiment show that the dimension of the tardive dyskinetic finger tremor time series is systematically lower than that evident in normal finger tremor. Furthermore, the variability of finger motion in both groups is inversely related to the dimension of the respective attractor dynamic. The neuroleptic medication appears to constrain the degrees of freedom regulated in organization of the motor system. (c) 1995 American Institute of Physics. PMID- 12780154 TI - Tremor classification and tremor time series analysis. AB - The separation between physiologic tremor (PT) in normal subjects and the pathological tremors of essential tremor (ET) or Parkinson's disease (PD) was investigated on the basis of monoaxial accelerometric recordings of 35 s hand tremor epochs. Frequency and amplitude were insufficient to separate between these conditions, except for the trivial distinction between normal and pathologic tremors that is already defined on the basis of amplitude. We found that waveform analysis revealed highly significant differences between normal and pathologic tremors, and, more importantly, among different forms of pathologic tremors. We found in our group of 25 patients with PT and 15 with ET a reasonable distinction with the third momentum and the time reversal invariance. A nearly complete distinction between these two conditions on the basis of the asymmetric decay of the autocorrelation function. We conclude that time series analysis can probably be developed into a powerful tool for the objective analysis of tremors. (c) 1995 American Institute of Physics. PMID- 12780155 TI - Variations on tremor parameters. AB - This paper describes our analysis procedure for long-term tremor EMG recordings, as well as three examples of applications. The description of the method focuses on how characteristics of the tremor (e.g. frequency, intensity, agonist antagonist interaction) can be defined and calculated based on surface EMG data. The resulting quantitative characteristics are called "tremor parameters." We discuss sinusoidally modulated, band-limited white noise as a model for pathological tremor-EMG, and show how the basic parameters can be extracted from this class of signals. The method is then applied to (1) estimate tremor severity in clinical studies, (2) quantify agonist-antagonist interaction, and (3) investigate the variations of the tremor parameters using simple methods from time-series analysis. (c) 1995 American Institute of Physics. PMID- 12780156 TI - Upright, correlated random walks: A statistical-biomechanics approach to the human postural control system. AB - The task of maintaining erect stance involves a complex sensorimotor control system, the output of which can be highly irregular. Even when a healthy individual attempts to stand still, the center of gravity of his or her body and the center of pressure (COP) under his or her feet continually move about in an erratic fashion. In this study, we approach the problem of characterizing postural sway from the perspective of random-walk theory. Specifically, we analyze COP trajectories as one-dimensional and two-dimensional random walks. These analyses reveal that over short-term intervals of time during undisturbed stance the COP behaves as a positively correlated random walk, whereas over long term intervals of time it resembles a negatively correlated random walk. We interpret this novel finding as an indication that during quiet standing the postural control system utilizes open-loop and closed-loop control schemes over short-term and long-term intervals, respectively. From this perspective, our approach, known as stabilogram-diffusion analysis, has the advantage that it leads to the extraction of COP parameters which can be directly related to the steady-state behavior and functional interaction of the neuromuscular mechanisms underlying the maintenance of erect stance. (c) 1995 American Institute of Physics. PMID- 12780157 TI - Self-organizing dynamics of the human brain: Critical instabilities and Sil'nikov chaos. AB - Using a sensorimotor coordination task in conjunction with an array of SQUIDs (Superconducting QUantum Interference Devices) we demonstrate critical instabilities in human brain activity patterns. Analysis of the dominant spatial pattern of the brain and its time-varying amplitude displays a task-dependent geometry characteristic of Sil'nikov-like chaos, which changes qualitatively at the transition. (c) 1995 American Institute of Physics. PMID- 12780158 TI - Characterizing the dynamics of auditory perception. AB - After listening to a sound that is presented repeatedly, subjects report hearing different transforms of the original sound. The frequency of reported transforms is a sensitive index of some speech disorders as well as cognitive flexibility in aging. In this paper, we propose and investigate quantitative measures that characterize the dynamics of this phenomenon, known as the verbal transformation effect. In particular, we show that the distribution of the dwell time, the time spent perceiving a string of a given phonemic form before switching to another form, obeys a power law for normal subjects with an exponent valued between 1 and 2. This result suggests that within this paradigm there is no characteristic time scale for the perceptual process. Additionally, we analyze the correlation properties of the transforms. We suggest that the complexity measures and techniques introduced here might be useful diagnostic tools for a number of speech and cognitive disorders. (c) 1995 American Institute of Physics. PMID- 12780159 TI - Nonlinear dynamics in pulsatile secretion of parathyroid hormone in normal human subjects. AB - In many biological systems, information is transferred by hormonal ligands, and it is assumed that these hormonal signals encode developmental and regulatory programs in mammalian organisms. In contrast to the dogma of endocrine homeostasis, it could be shown that the biological information in hormonal networks is not only present as a constant hormone concentration in the circulation pool. Recently, it has become apparent that hormone pulses contribute to this hormonal pool, which modulates the responsiveness of receptors within the cell membrane by regulation of the receptor synthesis, movement within the membrane layer, coupling to signal transduction proteins and internalization. Phase space analysis of dynamic parathyroid hormone (PTH) secretion allowed the definition of a (in comparison to normal subjects) relatively quiet "low dynamic" secretory pattern in osteoporosis, and a "high dynamic" state in hyperparathyroidism. We now investigate whether this pulsatile secretion of PTH in healthy men exhibits characteristics of nonlinear determinism. Our findings suggest that this is conceivable, although on the basis of presently available data and techniques, no proof can be established. Nevertheless, pulsatile secretion of PTH might be a first example of nonlinear deterministic dynamics in an apparently irregular hormonal rhythm in human physiology. (c) 1995 American Institute of Physics. PMID- 12780160 TI - Quantitative analysis of heart rate variability. AB - In the modern industrialized countries every year several hundred thousands of people die due to sudden cardiac death. The individual risk for this sudden cardiac death cannot be defined precisely by common available, noninvasive diagnostic tools like Holter monitoring, highly amplified ECG and traditional linear analysis of heart rate variability (HRV). Therefore, we apply some rather unconventional methods of nonlinear dynamics to analyze the HRV. Especially, some complexity measures that are based on symbolic dynamics as well as a new measure, the renormalized entropy, detect some abnormalities in the HRV of several patients who have been classified in the low risk group by traditional methods. A combination of these complexity measures with the parameters in the frequency domain seems to be a promising way to get a more precise definition of the individual risk. These findings have to be validated by a representative number of patients. (c) 1995 American Institute of Physics. PMID- 12780161 TI - Nonlinear time series analysis of electrocardiograms. AB - In recent years there has been an increasing number of papers in the literature, applying the methods and techniques of Nonlinear Dynamics to the time series of electrical activity in normal electrocardiograms (ECGs) of various human subjects. Most of these studies are based primarily on correlation dimension estimates, and conclude that the dynamics of the ECG signal is deterministic and occurs on a chaotic attractor, whose dimension can distinguish between healthy and severely malfunctioning cases. In this paper, we first demonstrate that correlation dimension calculations must be used with care, as they do not always yield reliable estimates of the attractor's "dimension." We then carry out a number of additional tests (time differencing, smoothing, principal component analysis, surrogate data analysis, etc.) on the ECGs of three "normal" subjects and three "heavy smokers" at rest and after mild exercising, whose cardiac rhythms look very similar. Our main conclusion is that no major dynamical differences are evident in these signals. A preliminary estimate of three to four basic variables governing the dynamics (based on correlation dimension calculations) is updated to five to six, when temporal correlations between points are removed. Finally, in almost all cases, the transition between resting and mild exercising seems to imply a small increase in the complexity of cardiac dynamics. (c) 1995 American Institute of Physics. PMID- 12780162 TI - Age-related changes in the "complexity" of cardiovascular dynamics: A potential marker of vulnerability to disease. AB - Healthy physiologic control of cardiovascular function is a result of complex interactions between multiple regulatory processes that operate over different time scales. These include the sympathetic and parasympathetic nervous systems which regulate beat-to-beat heart rate (HR) and blood pressure (BP), as well as extravascular volume, body temperature, and sleep which influence HR and BP over the longer term. Interactions between these control systems generate highly variable fluctuations in continuous HR and BP signals. Techniques derived from nonlinear dynamics and chaos theory are now being adapted to quantify the dynamic behavior of physiologic time series and study their changes with age or disease. We have shown significant age-related changes in the 1/f(x) relationship between the log amplitude and log frequency of the heart rate power spectrum, as well as declines in approximate dimension and approximate entropy of both heart rate and blood pressure time series. These changes in the "complexity" of cardiovascular dynamics reflect the breakdown and decoupling of integrated physiologic regulatory systems with aging, and may signal an impairment in cardiovascular ability to adapt to external and internal perturbations. Studies are currently underway to determine whether the complexity of HR or BP time series can distinguish patients with fainting spells due to benign vasovagal reactions from those due to life-threatening cardiac arrhythmias. Thus, measures of the complexity of physiologic variability may provide novel methods to monitor cardiovascular aging and test the efficacy of specific interventions to improve adaptive capacity in old age. (c) 1995 American Institute of Physics. PMID- 12780163 TI - Approximate entropy (ApEn) as a complexity measure. AB - Approximate entropy (ApEn) is a recently developed statistic quantifying regularity and complexity, which appears to have potential application to a wide variety of relatively short (greater than 100 points) and noisy time-series data. The development of ApEn was motivated by data length constraints commonly encountered, e.g., in heart rate, EEG, and endocrine hormone secretion data sets. We describe ApEn implementation and interpretation, indicating its utility to distinguish correlated stochastic processes, and composite deterministic/ stochastic models. We discuss the key technical idea that motivates ApEn, that one need not fully reconstruct an attractor to discriminate in a statistically valid manner-marginal probability distributions often suffice for this purpose. Finally, we discuss why algorithms to compute, e.g., correlation dimension and the Kolmogorov-Sinai (KS) entropy, often work well for true dynamical systems, yet sometimes operationally confound for general models, with the aid of visual representations of reconstructed dynamics for two contrasting processes. (c) 1995 American Institute of Physics. PMID- 12780164 TI - Tests for nonlinearity in short stationary time series. AB - To compare direct tests for detecting determinism in chaotic time series, data from Henon, Lorenz, and Mackey-Glass equations were contaminated with various levels of additive colored noise. These data were analyzed with a variety of recently developed tests for determinism, and the results compared. (c) 1995 American Institute of Physics. PMID- 12780165 TI - Interspike interval embedding of chaotic signals. AB - According to a theorem of Takens [Lecture Notes in Mathematics (Springer-Verlag, Berlin, 1981), Vol. 898], dynamical state information can be reproduced from a time series of amplitude measurements. In this paper we investigate whether the same information can be reproduced from interspike interval (ISI) measurements. Assuming an integrate-and-fire model coupling the dynamical system to the spike train, there is a one-to-one correspondence between the system states and interspike interval vectors of sufficiently large dimension. The correspondence implies in particular that a data series of interspike intervals, formed in this manner, can be forecast from past history. This capability is demonstrated using a nonlinear prediction algorithm, and is found to be robust to noise. A set of interspike intervals measured from a simple neuronal circuit is studied for deterministic structure using a prediction error statistic. (c) 1995 American Institute of Physics. PMID- 12780166 TI - Noise in chaotic data: Diagnosis and treatment. AB - A prominent limiting factor in the analysis of chaotic time series are measurement errors in the data. We show that this influence can be quite severe, depending on the nature of the noise, the complexity of the signal, and on the application one has in mind. Theoretical considerations yield general upper bounds on the tolerable noise level for dimension, entropy and Lyapunov estimates. We discuss methods to detect and analyze the noise present in a measured data set. We show how the situation can be improved by nonlinear noise reduction. (c) 1995 American Institute of Physics. PMID- 12780167 TI - Dimension estimates and physiological data. AB - Dimension estimates for data from physiological systems are notoriously difficult since the data are far from ideal in the sense of deterministic dynamical systems. Possible pitfalls and necessary precautions are pointed out and a recipe is given which is viable for those researchers who want to use the Grassberger Procaccia algorithm but who are not familiar with the vast existing literature on dimension estimates. The relevance of dimension estimates for the characterization of physiological data is discussed, where both the cases of finding and not finding a low dimension are considered. (c) 1995 American Institute of Physics. PMID- 12780168 TI - A basic mathematical model of the immune response. AB - Interaction of the immune system with a target population of, e.g., bacteria, viruses, antigens, or tumor cells must be considered as a dynamic process. We describe this process by a system of two ordinary differential equations. Although the model is strongly idealized it demonstrates how the combination of a few proposed nonlinear interaction rules between the immune system and its targets are able to generate a considerable variety of different kinds of immune responses, many of which are observed both experimentally and clinically. In particular, solutions of the model equations correspond to states described by immunologists as "virgin state," "immune state" and "state of tolerance." The model successfully replicates the so-called primary and secondary response. Moreover, it predicts the existence of a threshold level for the amount of pathogen germs or of transplanted tumor cells below which the host is able to eliminate the infectious organism or to reject the tumor graft. We also find a long time coexistence of targets and immune competent cells including damped and undamped oscillations of both. Plausibly the model explains that if the number of transformed cells or pathogens exeeds definable values (poor antigenicity, high reproduction rate) the immune system fails to keep the disease under control. On the other hand, the model predicts apparently paradoxical situations including an increased chance of target survival despite enhanced immune activity or therapeutically achieved target reduction. A further obviously paradoxical behavior consists of a positive effect for the patient up to a complete cure by adding an additional target challenge where the benefit of the additional targets depends strongly on the time point and on their amount. Under periodically pulsed stimulation the model may show a chaotic time behavior of both target growth and immune response. (c) 1995 American Institute of Physics. PMID- 12780169 TI - Shift of a limit cycle in biology: From pathological to physiological homeostasia@f| AB - Biological systems may show homeostatic behaviors that are similar to the ones of forced dynamic systems with a stable limit cycle. For a large class of dynamic systems, it is shown that a shift of a pathological limit cycle over the physiological limit cycle can never be executed by means of a control with a desired periodicity. The above statement shows that the only possibility is to reduce as much as possible the dimensions of a small residual limit cycle. Moreover, it is possible to give some information about the structure of feedback laws that would allow the shift of the limit cycle. The fact that it is generally not possible to recover a physiological limit cycle from a pathological one, results into the fear of never or hardly ever reaching a physiological behavior, and it seems that any hope of therapeutics is given up. This leads to introduce the locking concept, which permits system parameters to change and provides the basis for an adaptive and iterative control, which allows a step by step approach and to finally reach the physiological limit cycle. (c) 1995 American Institute of Physics. PMID- 12780170 TI - "Dynamical confinement" in neural networks and cell cycle. AB - In this paper randomization of well-known former mathematical models is proposed (i.e., the Hopfield model for neural networks and the Hahn model for the cell cycle) in order to facilitate the study of their asymptotic behavior: in fact, we replace the determination of the stability basins for attractors and boundaries by the study of a unique (or a small number of) invariant measure(s), whose distribution function maxima (or, respectively, percentile contour lines) correspond to the location of the attractors (or, respectively, boundaries of their stability basins). We give the name of "confinement" to this localization of the mass of the invariant measure(s). We intend to show here that the study of the confinement is in certain cases easier than the study of underlying attractors, in particular if these last are numerous and possess small stability basins (for example, for the first time we calculate the invariant measure in the random Hopfield model in a case for which the deterministic version exhibits many attractors, and in a case of phase transition). (c) 1995 American Institute of Physics. PMID- 12780171 TI - A minimal single-channel model for the regularity of beating in the sinoatrial node. AB - It has been suggested that the normal irregular beating of the heart is a manifestation of deterministically chaotic dynamics. Evidence proffered in support of this hypothesis includes a 1/f-like power spectrum, a small noninteger correlation dimension, and self-similarity of the time series. The major cause of the normal fluctuations in heart rate is the impingement of several neural and hormonal control systems upon the sinoatrial node, the natural pacemaker of the heart. However, intrinsic fluctuations of beat rate can be seen in the isolated node, devoid of all neural and hormonal inputs, and even in a single cell isolated from the node. The electrical activity in such a single cell is generated by ions flowing through discrete channels in the cell membrane.We decided to test the hypothesis that the fluctuations in beat rate in a single cell might be due to the fluctuations in the activity of this population of single channels. We thus assemble a model consisting of 6000 channels and probe its dynamics. Each channel has one or more gates, all of which must be open to allow current to flow through the channel. Since these gates are thought to open and close in a random manner, we model each gate by a Markov process, assigning a pseudorandom number to each gate every time that it changes state from open to closed or vice versa. This number, in conjunction with the classical voltage dependent Hodgkin-Huxley-like rate constants that control the speed with which a gate will open or close, then determines when that gate will next change state. We also employ a second method that is much more efficient computationally, in which one computes the lifetime of the ensemble of 6000 channels. We show that the Monte Carlo model has behavior consistent with the hypothesis that the irregular beating seen experimentally in single nodal cells is due to the (pseudo)random opening and closing of single channels. However, since the pseudorandom number generator used in the simulations is deterministic, one cannot state that the activity in the model is random (or stochastic). Thus, it would be premature to claim that the irregularity of beating in a single nodal cell is accounted for by the stochastic behavior of a population of a few thousand single channels lying in the membrane of the cell. Finally, we consider some implications of our work for the naturally occurring in situ fluctuations in heart rate ("heart rate variability"). (c) 1995 American Institute of Physics. PMID- 12780172 TI - Phase resetting and dynamics in isolated atrioventricular nodal cell clusters. AB - In the heart, the AV node is the primary conduction pathway between the atria and ventricles and subserves an important function by virtue of its rate-dependent properties. Cell clusters isolated from the rabbit atrioventricular (AV) node beat with a stable rhythm (cycle length: 300-520 ms) and are characterized by slow action potential upstroke velocities (7 to 30 V/s). The goal of this study is to better characterize the phase resetting and the rhythms during periodic stimulation of this slow inward current system. Single or periodic depolarizing pulses (20 ms in duration) were injected into AV nodal cell clusters using glass microelectrodes. Phase resetting curves of both strong, weak as well as discontinuous types were obtained by applying single current pulses of different intensities and latencies following every ten action potentials. Graded responses were elicited in a wide range of stimulus phases and amplitudes. A single premature stimulus caused a transient prolongation of the cycle length. Sustained periodic stimulation, at rates faster than the intrinsic beat rate, resulted in various N:M (stimulus frequency: action potential frequency) entrainment rhythms as well as periodic or irregular changes in action potential morphology. The changes in action potential characteristics were evaluated by computing the area under the action potential trace and above a fixed threshold (-45 mV). We show that the variations in action potential morphology play a major role in the onset of complicated dynamics observed in this experimental preparation. In this context, the prediction of entrainment rhythms using techniques based on the iteration of phase resetting curves (PRCs) is inadequate since the PRC does not carry information directly related to the changes in action potential morphology. This study demonstrates the need to consider graded events which, though not propagated, have important implications in the understanding of dynamical diseases of the heart. (c) 1995 American Institute of Physics. PMID- 12780173 TI - Phase-locking regions in a forced model of slow insulin and glucose oscillations. AB - We present a detailed numerical investigation of the phase-locking regions in a forced model of slow oscillations in human insulin secretion and blood glucose concentration. The bifurcation structures of period 2pi and 4pi tongues are mapped out and found to be qualitatively identical to those of several other periodically forced self-oscillating systems operating across a Hopf-bifurcation point. The numerical analyses are supplemented by clinical experiments. (c) 1995 American Institute of Physics. PMID- 12780174 TI - Discrete-time stimulation of the oscillatory and excitable forms of a FitzHugh Nagumo model applied to the pulsatile release of luteinizing hormone releasing hormone. AB - A model for the pulsatile release of luteinizing hormone releasing hormone (LHRH) can be reduced to a FitzHugh-Nagumo model subject to regular and quasiregular (i.e., with slight random variation in the interstimulus interval), discrete-time stimulation. The relationship of output pulse frequency (OPF) to stimulus frequency is compared between the excitable and oscillatory forms of the model and discussed in the context of results from other pulse-driven model systems. Some examples of the changes in OPF caused by quasiregular and purely Poissonian stimuli are given for the excitable case. The unstimulated system frequently interacts with the stimulation in such a complex manner that the OPF bears little resemblance to the frequency of stimulation or of the unstimulated system. Furthermore, the inability of the oscillatory form of the model to allow complete suppression of output pulses for moderate stimulation frequencies suggests that the LHRH system can be more appropriately described by the excitable form of the model. (c) 1995 American Institute of Physics. PMID- 12780175 TI - Mechanisms of stochastic phase locking. AB - Periodically driven nonlinear oscillators can exhibit a form of phase locking in which a well-defined feature of the motion occurs near a preferred phase of the stimulus, but a random number of stimulus cycles are skipped between its occurrences. This feature may be an action potential, or another crossing by a state variable of some specific value. This behavior can also occur when no apparent external periodic forcing is present. The phase preference is then measured with respect to a time scale internal to the system. Models of these behaviors are briefly reviewed, and new mechanisms are presented that involve the coupling of noise to the equations of motion. Our study investigates such stochastic phase locking near bifurcations commonly present in models of biological oscillators: (1) a supercritical and (2) a subcritical Hopf bifurcation, and, under autonomous conditions, near (3) a saddle-node bifurcation, and (4) chaotic behavior. Our results complement previous studies of aperiodic phase locking in which noise perturbs deterministic phase-locked motion. In our study however, we emphasize how noise can induce a stochastic phase-locked motion that does not have a similar deterministic counterpart. Although our study focuses on models of excitable and bursting neurons, our results are applicable to other oscillators, such as those discussed in the respiratory and cardiac literatures. (c) 1995 American Institute of Physics. PMID- 12780176 TI - Checkerboard maps. AB - When a map has one positive Lyapunov exponent, its attractors often look like multidimensional, Cantorial plates of spaghetti. What saves the situation is that there is a deterministic jumping from strand to strand. We propose to approximate such attractors as finite sets of K suitably prescribed curves, each parametrized by an interval. The action of the map on each attractor is then approximated by a map that takes a set of curves into itself, and we graph it on a KxK checkerboard as a discontinuous one-dimensional map that captures the quantitative dynamics of the original system when K is sufficiently large. (c) 1995 American Institute of Physics. PMID- 12780177 TI - On the singularity structure of invariant curves of symplectic mappings. AB - We study invariant curves in standard-like maps conjugate to rigid rotation with complex frequencies. The main goal is to study the analyticity domain of the functions defined perturbatively by Lindstedt perturbation expansions. We argue, based on infinite-dimensional bifurcation theory that the boundary of analyticity should typically consist of branch points of order two and we verify it in some examples using nonperturbative numerical methods. We show that this nature of the singularities of the analyticity domain can explain previously reported numerical results and also suggests other numerical methods. (c) 1995 American Institute of Physics. PMID- 12780178 TI - Statistical properties of 2-D generalized hyperbolic attractors. AB - Recently Pesin introduced a large class of hyperbolic attractors, and for those attractors he established the Smale spectral decomposition. In this paper our main results are a stretched exponential bound on the decay of correlations and the central limit theorem. Also we will obtain conditions under which two well known attractors-those of Belykh and Lozi-are subject to our main results. (c) 1995 American Institute of Physics. PMID- 12780179 TI - Characterizing strange nonchaotic attractors. AB - Strange nonchaotic attractors typically appear in quasiperiodically driven nonlinear systems. Two methods of their characterization are proposed. The first one is based on the bifurcation analysis of the systems, resulting from periodic approximations of the quasiperiodic forcing. Second, we propose to characterize their strangeness by calculating a phase sensitivity exponent, that measures the sensitivity with respect to changes of the phase of the external force. It is shown that phase sensitivity appears if there is a nonzero probability for positive local Lyapunov exponents to occur. (c) 1995 American Institute of Physics. PMID- 12780180 TI - Periodic orbits of nonscaling Hamiltonian systems from quantum mechanics. AB - Quantal (E,tau) plots are constructed from the eigenvalues of the quantum system. We demonstrate that these representations display the periodic orbits of the classical system, including bifurcations and the transition from stable to unstable. (c) 1995 American Institute of Physics. PMID- 12780181 TI - Unstable periodic orbits and templates of the Rossler system: Toward a systematic topological characterization. AB - The Rossler system has been exhaustively studied for parameter values (a in [0.33,0.557],b=2,c=4). Periodic orbits have been systematically extracted from Poincare maps and the following problems have been addressed: (i) all low order periodic orbits are extracted, (ii) encoding of periodic orbits by symbolic dynamics (from 2 letters up to 11 letters) is achieved, (iii) some rules of growth and of pruning of the periodic orbits population are obtained, and (iv) the templates of the attractors are elaborated to characterize the attractors topology. (c) 1995 American Institute of Physics. PMID- 12780182 TI - Time scale to ergodicity in the Fermi-Pasta-Ulam system. AB - We study the approach to near-equipartition in the N-dimensional Fermi-Pasta-Ulam Hamiltonian with quartic (hard spring) nonlinearity. We investigate numerically the time evolution of orbits with initial energy in some few low-frequency linear modes. Our results indicate a transition where, above a critical energy which is independent of N, one can reach equipartition if one waits for a time proportional to N(2). Below this critical energy the time to equipartition is exponentially long. We develop a theory to determine the time evolution and the excitation of the nonlinear modes based on a resonant normal form treatment of the resonances among the oscillators. Our theory predicts the critical energy for equipartition, the time scale to equipartition, and the form of the nonlinear modes below equipartition, in qualitative agreement with the numerical results. (c) 1995 American Institute of Physics. PMID- 12780183 TI - Distinguishing the transition to chaos in a spherical pendulum. AB - Complex responses are studied for a spherical pendulum whose support is excited with a translational periodic motion. Governing equations are studied analytically to allow prediction of responses under various excitation conditions. Stability for certain cases of damping is predicted by means of existing analysis and compared with experimental data. Numerical time-step integration of the governing equations is developed to predict responses for various types of excitation and damping conditions. Predicted results are compared with corresponding motions measured in an experimental spherical pendulum system. A data acquisition system is included whereby detailed digitized time histories of the pendulum motion can be established and various parameters can be computed to characterize the type of motion present. Two new vector spaces are defined for describing complex responses which occur for certain specified excitation conditions. It is shown in these parameter spaces that the transition from quasiperiodic to chaotic motions can be carefully quantified in systems with very light damping. This discovery provides a convenient means for comparison of complex motions in the numerical and experimental air pendulum systems. The implications of the results are important for dynamic response in various applications, including fluid motions in satellite tanks and other nonlinear time dependent physical processes which include very light damping. (c) 1995 American Institute of Physics. PMID- 12780184 TI - Nonlinear dynamical analysis of turbulence in a stable cloud layer. AB - An eight mode truncated spectral model based on Burgers' approximation to the one dimensional Navier-Stokes equations is used to compute the Lyapunov dimension of the dynamical attractor for turbulence in a stable cloud layer. The model results are compared with the correlation dimension obtained earlier from a time series of radar Doppler and reflectivity signals from a turbulent layer in a marine stratus cloud. The analysis supports a weak coupling explanation for the lower correlation dimension found for the reflectivity time series compared with that for the Doppler time series. Turbulent Prandtl number emerges from the analysis as a flow parameter which can enlarge the dimension of the model's dynamical attractor, but the attractor dimension computed for the model remains lower than the radar Doppler correlation dimension. Linear stability analysis of the model's equilibrium states suggests that a nontruncated version of the model will possess an attractor which is also of lower dimension than the radar Doppler correlation dimension. (c) 1995 American Institute of Physics. PMID- 12780185 TI - Effect of a magnetic flux line on the quantum beats in the Henon-Heiles level density. AB - The quantum density of states of the Henon-Heiles potential displays a pronounced beating pattern. This has been explained by the interference of three isolated classical periodic orbits with nearby actions and periods. A singular magnetic flux line, passing through the origin, drastically alters the beats even though the classical Lagrangian equations of motion remain unchanged. Some of the changes can be easily understood in terms of the Aharonov-Bohm effect. However, we find that the standard periodic orbit theory does not reproduce the diffraction-like quantum effects on those classical orbits which intersect the singular flux line, and argue that corrections of relative order variant Planck's over 2pi are necessary to describe these effects. We also discuss the changes in the distribution of nearest-neighbor spacings in the eigenvalue spectrum, brought about by the flux line. (c) 1995 American Institute of Physics. PMID- 12780186 TI - Finite-dimensional behavior in dissipative partial differential equations. AB - Dissipative partial differential equations have applications throughout the sciences: models of turbulence in fluids, chemical reactions, and morphogenesis in biology can all be written in a general form which allows them to be subjected to a unified analysis. Recent results on these equations show that in many cases they are not as complex as they initially appear, and can be converted into a set of ordinary differential equations. However, most of the relevant references present a bewildering array of terms which can obscure the simple underlying ideas. The main purpose of this paper is to introduce this terminology, motivated by several major results, slowly and by example. Detailed proofs are omitted, but it is hoped that this approach will give a good understanding of and intuitive feel for the subject without recourse to technicalities. Nevertheless, sufficient mathematical detail is included to allow application of these results to many examples. (c) 1995 American Institute of Physics. PMID- 12780187 TI - Erratum: "Torus knots and polynomial invariants for a class of soliton equations" [Chaos 3, 83 (1993)]. PMID- 12780188 TI - Variational principle for periodic trajectories of hyperbolic billiards. AB - We prove for some classes of hyperbolic billiards that the action functional has only one local minimum or only one local maximum for any finite admissible sequence of regular components of the boundary. This result suggests an effective algorithm for the search of all periodic trajectories of these billiards. (c) 1995 American Institute of Physics. PMID- 12780189 TI - Monitoring changes in time of chaotic nonlinear systems. AB - We extend our method for classifying signals from chaotic nonlinear dynamical systems to the problem of monitoring chaotic nonlinear dynamical systems with the goal of detecting that the state of a system has changed. One potential application would be to systems where the changes are not easily detectable by spectral analysis or other linear techniques. The method is expected to be most useful in comparison to other techniques when there are other signals or noise present, some of which have a broad band frequency spectrum, and the signal of interest is associated with either a low dimensional dynamical system or a low dimensional chaotic attractor. The method is applied to data from a laboratory model of a fluidized bed reactor and to data from a gyroscope as well as to numerically generated signals from mathematical models. For the dynamical systems considered in the paper, the proposed method provides significantly better discrimination than spectral analysis. (c) 1995 American Institute of Physics. PMID- 12780190 TI - Bifurcations in biparametric quadratic potentials. AB - Numerous dynamical systems are represented by quadratic Hamiltonians with the phase space on the S (2) sphere. For a class of these Hamiltonians depending on two parameters, we analyze the occurrence of bifurcations and we obtain the bifurcation lines in the parameter plane. As the parameters evolve, the appearance-disappearance of homoclinic orbits in the phase portrait is governed by three types of bifurcations, the pitchfork, the teardrop and the oyster bifurcations. We find that the teardrop bifurcation is associated with a non elementary fixed point whose Poincare index is zero. (c) 1995 American Institute of Physics. PMID- 12780191 TI - Logarithmic correction to the probability of capture for dissipatively perturbed Hamiltonian systems. AB - Hamiltonian systems are analyzed with a double homoclinic orbit connecting a saddle to itself. Competing centers exist. A small dissipative perturbation causes the stable and unstable manifolds of the saddle point to break apart. The stable manifolds of the saddle point are the boundaries of the basin of attraction for the competing attractors. With small dissipation, the boundaries of the basins of attraction are known to be tightly wound and spiral-like. Small changes in the initial condition can alter the equilibrium to which the solution is attracted. Near the unperturbed homoclinic orbit, the boundary of the basin of attraction consists of a large sequence of nearly homoclinic orbits surrounded by close approaches to the saddle point. The slow passage through an unperturbed homoclinic orbit (separatrix) is determined by the change in the value of the Hamiltonian from one saddle approach to the next. The probability of capture can be asymptotically approximated using this change in the Hamiltonian. The well known leading-order change of the Hamiltonian from one saddle approach to the next is due to the effect of the perturbation on the homoclinic orbit. A logarithmic correction to this change of the Hamiltonian is shown to be due to the effect of the perturbation on the saddle point itself. It is shown that the probability of capture can be significantly altered from the well-known leading order probability for Hamiltonian systems with double homoclinic orbits of the twisted type, an example of which is the Hamiltonian system corresponding to primary resonance. Numerical integration of the perturbed Hamiltonian system is used to verify the accuracy of the analytic formulas for the change in the Hamiltonian from one saddle approach to the next. (c) 1995 American Institute of Physics. PMID- 12780192 TI - Secondary homoclinic bifurcation theorems. AB - We develop criteria for detecting secondary intersections and tangencies of the stable and unstable manifolds of hyperbolic periodic orbits appearing in time periodically perturbed one degree of freedom Hamiltonian systems. A function, called the "Secondary Melnikov Function" (SMF) is constructed, and it is proved that simple (resp. degenerate) zeros of this function correspond to transverse (resp. tangent) intersections of the manifolds. The theory identifies and predicts the rotary number of the intersection (the number of "humps" of the homoclinic orbit), the transition number of the homoclinic points (the number of periods between humps), the existence of tangencies, and the scaling of the intersection angles near tangent bifurcations perturbationally. The theory predicts the minimal transition number of the homoclinic points of a homoclinic tangle. This number determines the relevant time scale, the minimal stretching rate (which is related to the topological entropy) and the transport mechanism as described by the TAM, a transport theory for two-dimensional area-preserving chaotic maps. The implications of this theory on the study of dissipative systems have yet to be explored. (c) 1995 American Institute of Physics. PMID- 12780193 TI - Secondary cycle of equilibria in a system with cosymmetry, its creation by bifurcation and impossibility of symmetric treatment of it. AB - A study is reported of the bifurcation of a cycle of equilibria of an autonomous differential equation with cosymmetry in Hilbert space, which is a simulation of the problem of planar filtrational convection of a fluid in a porous medium. The Lyapunov-Schmidt method and perturbation theory are used to find its amplitude and the damping rate of the dominant mode. It is shown that, in the abstract general model, and also in the problem of convection in a rectangular container, this damping rate varies along the cycle of equilibria. Hence, the cycle of equilibria cannot be an orbit of the action of any symmetry group of the given system. (c) 1995 American Institute of Physics. PMID- 12780194 TI - Singular Lyapunov spectra and conservation laws. AB - We give analytic arguments and numerical evidence to show that the presence of conservation laws can produce a singularity in the spectrum of Lyapunov exponents for extended dynamical systems of low spatial dimensionality. This phenomenon can be used, e.g., for finding hidden conservation laws. (c) 1995 American Institute of Physics. PMID- 12780195 TI - On stability and instability criteria for magnetohydrodynamics. AB - It is shown that for most, but not all, three-dimensional magnetohydrodynamic (MHD) equilibria the second variation of the energy is indefinite. Thus the class of such equilibria whose stability might be determined by the so-called Arnold criterion is very restricted. The converse question, namely conditions under which MHD equilibria will be unstable is considered in this paper. The following sufficient condition for linear instability in the Eulerian representation is presented: The maximal real part of the spectrum of the MHD equations linearized about an equilibrium state is bounded from below by the growth rate of an operator defined by a system of local partial differential equations (PDE). This instability criterion is applied to the case of axisymmetric toroidal equilibria. Sufficient conditions for instability, stronger than those previously known, are obtained for rotating MHD. (c) 1995 American Institute of Physics. PMID- 12780196 TI - On small stationary localized solutions for the generalized 1-D Swift-Hohenberg equation. AB - We prove the existence of small localized stationary solutions for the generalized Swift-Hohenberg equation and find under some assumption a part of a boundary of their existence in the parameter plane. The related stationary equation creates a reversible Hamiltonian system with two degrees of freedom that undergoes the Hamiltonian-Hopf bifurcation with an additional degeneracy. We investigate this bifurcation in a two-parameter unfolding by means of the sixth order normal form for the related Hamiltonian. The region where no localized solutions exist has been pointed out as well. (c) 1995 American Institute of Physics. PMID- 12780197 TI - Chaotic advection in a 2-D mixed convection flow. AB - Two-dimensional numerical simulations of particle advection in a channel flow with spatially periodic heating have been carried out. The velocity field is found to be periodic above a critical Rayleigh number of around 18 000 and a Reynolds number of 10. Particle motion becomes chaotic in the lower half plane almost immediately after this critical value is surpassed, as characterized by the power spectral density and Poincare section of the flow. As the Rayleigh number is increased further, particle motion in the entire domain becomes chaotic. (c) 1995 American Institute of Physics. PMID- 12780198 TI - The dynamics of a generalized Heisenberg ferromagnetic spin chain. AB - We introduce a generalized Heisenberg ferromagnetic spin chain with a four dimensional target space, and investigate its continuous limit, the generalized continuous Heisenberg model (GCHM). We reduce the dynamics of the GCHM to a nonlinear evolution of space curves in four dimensions. The space curve evolution is expressed in terms of a system of coupled nonlinear equations for the three curvatures, k(1)(x),k(2)(x),k(3)(x), of a curve in R(4). Applying the Painleve analysis to the stationary equations, we conclude that GCHM, in general, is not integrable, unless k(1) is constant. We obtain explicit solutions of the resulting stationary system under the latter condition. (c) 1995 American Institute of Physics. PMID- 12780199 TI - Involutes: the geometry of chemical waves rotating in annular membranes. AB - According to earlier theories certain parts of a chemical wave front propagating in a 2-D excitable medium with a convex obstacle should be involutes of that obstacle. The present paper discusses a special case where self-sustained chemical waves are rotating around a central obstacle in an annular 2-D excitable region. A simple geometrical model of wave propagation based on the Fermat principle (minimum propagation time) is suggested. Applying this model it is shown that the wave fronts in the case of an annular excitable region should be purely involutes of the central obstacle in the asymptotic state. This theory is supported by experiments in a novel membrane reactor where a catalyst of the Belousov-Zhabotinsky reaction is fixed on a porous membrane combined with a gel medium. Involutes of circular and triangular obstacles are observed experimentally. Deviations from the ideal involute geometry are explained by inhomogeneities in the membrane. (c) 1995 American Institute of Physics. PMID- 12780200 TI - Multiple time scale analysis of two models for the peroxidase-oxidase reaction. AB - A multiple time scale analysis of two four-variable models of the peroxidase oxidase reaction, the DOP, and the Olsen model, is carried out. It is shown that autonomous limit cycle oscillations are exhibited by the fast subsets of these two models, but only in certain regions of parameter space, confirming the prior suggestion that the slow variable (NADH) is not essential for oscillatory behavior. However, it is found that the slow variable is essential for oscillatory behavior over other ranges of parameter values, and is always essential for complex oscillatory and chaotic behavior. This latter conclusion is based on a study involving driving the fast subset with a sinusoidally varying (NADH). This study suggests the level of coupling between fast and slow variables of an autonomous system necessary to cause the chaos observed in the DOP model. Further study of the driven system allows for the identification of a natural period of the nonoscillatory but bistable fast subsystem and a set of rules for applying a parametric driving in such a way as to generate a more complete Farey sequence from a truncated Farey sequence. These conclusions are used to compare the very similar DOP and Olsen models, which, nevertheless, exhibit quite different Farey sequences and routes to chaos. (c) 1995 American Institute of Physics. PMID- 12780201 TI - Noise controlled spiral growth in excitable media. AB - We consider a two-dimensional pulse-coupled array of noisy threshold elements with a long-range interaction. For tight coupling, we observe the formation of spatio-temporal excitation waves, such as target and spiral waves. For weak coupling, we find noise sustained spiral growth, where the noise level controls the scale of the spiral. (c) 1995 American Institute of Physics. PMID- 12780202 TI - On the interplay between symmetry breaking, integrability, and chaos in the semiclassical limit of the Heisenberg system. AB - In this work we present a detailed numerical analysis of the interplay between symmetry breaking, integrability, and chaos in the two- and three-spin Heisenberg models. The results suggest that a very simple and powerful tool to convey such information are the plots of the energy level spacings Delta(n) versus the energy level index n, together with the correlation plots Delta(n+1)xDelta(n). When integrability is broken, these plots are shown to identify very sharply an energy below which one has chaotic behavior. The particularly strong point in favor of such analysis is that it can be useful in partially chaotic regimes. (c) 1995 American Institute of Physics. PMID- 12780203 TI - Quantum zeta function for perturbed cat maps. AB - The behavior of semiclassical approximations to the spectra of perturbed quantum cat maps is examined as the perturbation parameter brings the corresponding classical system into the nonhyperbolic regime. The approximations are initially accurate but large errors are found to appear in the traces and in the coefficients of the characteristic polynomial after nonhyperbolic structures appear. Nevertheless, the eigenvalues obtained from them remain accurate up to large perturbations. (c) 1995 American Institute of Physics. PMID- 12780204 TI - Introduction to acoustical chaos. PMID- 12780205 TI - Transitions and statistical characteristics of vibrations in a bimodular oscillator. AB - Regular and stochastic oscillations in a simple periodically forced vibroacoustic system with a piecewise-linear (bimodular) elasticity are considered from the viewpoint of their statistical properties: oscillation spectra, the largest Lyapunov exponents, and fractal dimension. It is shown that a strange attractor exists in a limited range of parameters together with a triple-period dynamic cycle being absorbed by the latter at some finite value of the parameter. The corresponding bifurcation process is followed in detail. (c) 1995 American Institute of Physics. PMID- 12780206 TI - Dynamical systems theory for music dynamics. AB - We show that, when music pieces are cast in the form of time series of pitch variations, the concepts and tools of dynamical systems theory can be applied to the analysis of temporal dynamics in music. (i) Phase space portraits are constructed from the time series wherefrom the dimensionality is evaluated as a measure of the global dynamics of each piece. (ii) Spectral analysis of the time series yields power spectra ( approximately f(-nu)) close to red noise (nu approximately 2) in the low frequency range. (iii) We define an information entropy which provides a measure of the local dynamics in the musical piece; the entropy can be interpreted as an evaluation of the degree of complexity in the music, but there is no evidence of an analytical relation between local and global dynamics. These findings are based on computations performed on eighty sequences sampled in the music literature from the 18th to the 20th century. (c) 1995 American Institute of Physics. PMID- 12780207 TI - The transient behaviour of models of bowed-string motion. AB - Theoretical models of the action of a bowed string may be able to shed light on differences of "playability" between different violins. Subjective judgements seem to be concerned, at least in part, with the robustness with which one particular oscillation regime of the string (the "Helmholtz motion") may be obtained under different bowing conditions. In this paper, after a review of bowed-string modelling, systematic simulation is used to obtain plots of the basin of attraction of the Helmholtz motion in a particular subspace of the player's control space. Variations in the size and structure of this basin of attraction are seen when parameters of the problem are varied, and some physical interpretation of these variations is given. Some parallels and contrasts are pointed out between the particular features of the bowed string as a nonlinear system, and the range of more familiar dynamical systems. (c) 1995 American Institute of Physics. PMID- 12780208 TI - The nonlinear decay of complex signals in dissipative media. AB - The solution of Burgers' equation with random initial conditions is often said to describe "Burgers turbulence." The Burgers equation describes two fundamental effects characteristic of any turbulence-the nonlinear transfer of energy over the spectrum and the dissipation of energy in the small-scale components. Strong interaction between coherent harmonics, associated with the nondispersive nature of the dynamics, leads to the appearance of local self-similar structure. In Burgers' equation, continuous random initial fields are transformed into sequences of regions with regular behavior, with random locations of the shocks separating them. Moreover, the statistical properties of such random fields are also self-similar. It is already known that the merging of the shocks leads to an increase of the external scale of the turbulence, and because of this the energy of a random signal ("noise") decreases more slowly than the energy of simple signals. Here we show that similar behavior takes place for complex regular signals with fractal structure in the coordinate or in the wave-number space. In all these cases, the law of increase of the external scale is determined by the behavior of the structure function of the integral of the initial field-i.e., the structure function of the initial action. (c) 1995 American Institute of Physics. PMID- 12780209 TI - Bifurcations in biparametric quadratic potentials. II. AB - Quadratic Hamiltonians with the phase space on the S (2) sphere represent numerous dynamical systems. There are only two classes of quadratic Hamiltonians depending on two parameters. We analyze the occurrence of bifurcations and we obtain the bifurcation lines in the parameter plane for one of these classes, thus complementing the work done in a previous paper where the other class was analyzed. As the parameters evolve, the appearance-disappearance of homoclinic orbits in the phase portrait is governed by four types of bifurcations: namely the pitchfork, the butterfly, the oyster and the pentadent bifurcations. We find also values where the system is degenerate, that is, there are nonisolated equilibria. (c) 1995 American Institute of Physics. PMID- 12780210 TI - The nonequilibrium Lorentz gas. AB - We study the conductivity of a Lorentz gas system, composed of a regular array of fixed scatterers and a point-like moving particle, as a function of the strength of an applied external field. In order to obtain a nonequilibrium stationary state, the speed of the point particle is fixed by the action of a Gaussian thermostat. For small fields the system is ergodic and the diffusion coefficient is well defined. We show that in this range the Periodic Orbit Expansion can be successfully applied to compute the values of the thermodynamic variables. At larger values of the field we observe a variety of possible dynamics, including the breakdown of ergodic behavior, and later the existence of a single stable trajectory for the largest fields. We also study the behavior of the system as a function of the orientation of the array of scatterers with respect to the external field. Finally, we present a detailed dynamical study of the transitions in the bifurcation sequence in both the elementary cell and the fundamental domain. The consequences of this behavior for the ergodicity of the system are explored. (c) 1995 American Institute of Physics. PMID- 12780211 TI - The iteration-approximation decoupling in the reversible KAM theory. AB - General theorems on the persistence of quasiperiodic motions in reversible flows and diffeomorphisms satisfying very weak nondegeneracy conditions are obtained by a new method. The essence of this method is that the reversible system under consideration is embedded in a multiparameter family of reversible systems, and standard results on Diophantine approximations of dependent quantities are then applied to Whitney-smooth Cantor foliations of invariant tori of this family. Invariant tori are constructed for all the permissible values of m, p, q (for vector fields V) or m, p, q, P, Q (for diffeomorphisms A) where m is the torus dimension, (q,p) is the type of the reversing involution G, and (Q,P) is the type of the involution AG. The excitation of elliptic normal modes is also considered. (c) 1995 American Institute of Physics. PMID- 12780212 TI - Fixed-point densities for a quasiperiodic kicked-oscillator map. AB - Suppose that a one-dimensional harmonic oscillator is subjected to instantaneous kicks q times per natural period, with the kick amplitude varying sinusoidally with position. Viewed stroboscopically in phase space, the motion has an infinitely extended periodic or quasiperiodic array of fixed points, as well as an infinite web of chaotic orbits. In the present work (restricted to the quasiperiodic case q=5) the fixed points are classified according to their local linear behavior, which depends essentially on a single variable, the residue R. With the aid of a five-dimensional embedding, a function rho(R) is calculated which for infinitesimal DeltaR gives the average density of fixed points in the plane with residue in the range (R,R+DeltaR). The location and strength of the singularities and discontinuities of rho(R) are extracted from relatively simple transcendental equations, and this makes possible efficient numerical determination of rho(R). An exact equality for the densities of positive-R and negative-R fixed points is proved using decagonal symmetry and the integral representation of rho(R). For parameter values below the period-doubling threshold, there are no unstable fixed points with R greater, similar 0, and so we have equality of the densities of stable centers and unstable saddles. (c) 1995 American Institute of Physics. PMID- 12780213 TI - The dynamics of spatiotemporal modulations. AB - The modulational instability of traveling waves is often thought to be a crucial point in the mechanism of transition to space-time disorder and turbulence. The aim of this paper is to study the effect of spatiotemporal modulations on some dynamics u(0)(x,t), which may occur as an instability process when a control parameter varies, for instance. We analyze the properties of the modulated dynamics of the form g(1)(x)g(2)(t)u(0)(x,t) compared to those of the reference dynamics u(0)(x,t), using operator theory. We show that, if the reference dynamics is invariant under some space-time symmetry in the sense of Ref. [J. Nonlinear Sci. 2, 183 (1992)], the modulation has the effect of either deforming this symmetry or breaking it, depending on whether the corresponding operator remains unitary or not. We also demonstrate that the smallest Euclidean space containing the modulated dynamics has a dimension smaller than or equal to the smallest Euclidean space containing u(0)(x,t). The previous results are then applied to the case of modulated uniformly traveling waves. While the spatiotemporal translation invariance of the wave never persists in the presence of a modulation, the existence of a spatiotemporal symmetry depends on the resonance of the Fourier sidebands due to the modulation. In case of nonresonance, a spatiotemporal symmetry exists and is explicitly determined. In this situation, the modulated wave and the carrier wave have the same spectrum (up to a normalization factor), the same entropy, and the spatial (resp., temporal) two-point correlation is deformed only by the spatial (resp., temporal) modulation. (c) 1995 American Institute of Physics. PMID- 12780214 TI - The evolution of chemical patterns in reactive liquids, driven by hydrodynamic instabilities. AB - We summarize our activity in unveiling a very wide phenomenon: When a chemical reaction takes place at a liquid interface, spectacular patterns of product form (see Plate 1). The pattern formation phenomenon is general, and is observed in reactions between liquids separated by a membrane, in liquids subjected to gaseous reactants, and in photoreactive liquids. We have demonstrated the phenomenon on over 100 different reactions of all types, thus discovering what we believe to be one of the widest macroscopic pattern formation processes known to chemistry. As can be seen in the accompanying pictures, the richness, beauty, and variations in types of patterns can be breathtaking. Two important aspects of these patterns are noted: First, the patterns are true far-from-equilibrium structures, which are maintained only as long as reactants are available, or only as long as light energy is supplied to the system; and second, the chemical products that form the patterns are not precipitates, but are entirely soluble in the liquid in which they form. Thus, if the containers in which the patterns form are shaken or stirred, a homogeneous solution results. Our research of this phenomenon concentrated on three main aspects. The first one was phenomenological. Here we explored the scope and generality of the phenomenon, motivated both by the aesthetic appeal of the phenomenon, and by the puzzle of how is it that such a wide-scope, experimentally simple phenomenon, has by and large, escaped the attention of the scientific community.The second aspect was devoted to the understanding of the underlying general mechanism. Of the many mechanisms we analyzed and tested, some very complex, others quite trivial, the one that fits the majority of the physical and chemical observations is the following: By performing a reaction through a liquid interface, a concentration gradient of the product forms near the interface. We have shown that in many cases, these gradients lead to hydrodynamic instabilities, which then break nonlinearly into a pattern which onsets slow convections. In other words, we found that these patterns mark the route along which a chemical instability relaxes. The third aspect of our research was theoretical. Here we concentrated in depth on one of the reactions (the Fe(+2)/Fe(+3) photoredox reaction), determined all its important physical parameters, and modeled its behavior theoretically. Our model, which was based on the instability buildup described above, was solved numerically, and its results compared with computerized image analysis of the evolving patterns; very good agreement between theory and experiment, was obtained. (c) 1995 American Institute of Physics. PMID- 12780215 TI - Kink dynamics in one-dimensional coupled map lattices. AB - We examine the problem of the dynamics of interfaces in a one-dimensional space time discrete dynamical system. Two different regimes are studied: the non propagating and the propagating one. In the first case, after proving the existence of such solutions, we show how they can be described using Taylor expansions. The second situation deals with the assumption of a travelling wave to follow the kink propagation. Then a comparison with the corresponding continuous model is proposed. We find that these methods are useful in simple dynamical situations but their application to complex dynamical behaviour is not yet understood. (c) 1995 American Institute of Physics. PMID- 12780216 TI - Correlation in stimulated respiratory neural noise. AB - Noise in spontaneous respiratory neural activity of the neonatal rat isolated brainstem-spinal cord preparation stimulated with acetylcholine (ACh) exhibits positive correlation. Neural activity from the C4 (phrenic) ventral spinal rootlet, integrated and corrected for slowly changing trend, is interpreted as a fractal record in time by rescaled range, relative dispersional, and power spectral analyses. The Hurst exponent H measured from time series of 64 consecutive signal levels recorded at 2 s intervals during perfusion of the preparation with artificial cerebrospinal fluid containing ACh at concentrations 62.5 to 1000 &mgr;M increases to a maximum of 0.875+/-0.087 (SD) at 250 &mgr;M ACh and decreases with higher ACh concentration. Corrections for bias in measurement of H were made using two different kinds of simulated fractional Gaussian noise. Within limits of experimental procedure and short data series, we conclude that in the presence of added ACh of concentration 250 to 500 &mgr;M, noise which occurs in spontaneous respiratory-related neural activity in the isolated brainstem-spinal cord preparation observed at uniform time intervals exhibits positive correlation. (c) 1995 American Institute of Physics. PMID- 12780217 TI - Signatures of deterministic chaos in radar sea clutter and ocean surface winds. AB - Several time series of wind components and X-band Doppler radar signals, gathered concurrently over a approximately 0.01 km(2) area of the ocean surface, were examined for evidence of a low-dimensional dynamical attractor with the Grassburger-Procaccia algorithm. Only the vertically polarized radar reflectivity and the horizontal surface wind speed time series suggested the presence of such an attractor. The correlation dimension for these two observables appeared to be nearly the same. This suggested a working hypothesis that the dynamical behavior of both the vertically polarized radar reflectivity and the horizontal surface winds are controlled by a single low-dimensional dynamical system. The hypothesis was further examined by predicting winds from radar reflectivity, using a neural network deterministic model, and comparing the prediction performance with that of the SEASAT statistical algorithm for retrieving surface winds from radar backscatter. It was found that the deterministic model did, in fact, achieve a higher prediction correlation coefficient for a limited time period. (c) 1995 American Institute of Physics. PMID- 12780218 TI - Unstable evolution of pointwise trajectory solutions to chaotic maps. AB - Simple chaotic maps are used to illustrate the inherent instability of trajectory solutions to the Frobenius-Perron equation. This is demonstrated by the difference in the behavior of delta-function solutions and of extended densities. Extended densities evolve asymptotically and irreversibly into invariant measures on stationary attractors. Pointwise trajectories chaotically roam over these attractors forever. Periodic Gaussian distributions on the unit interval are used to provide insight. Viewing evolving densities as ensembles of unstable pointwise trajectories gives densities a stochastic interpretation. (c) 1995 American Institute of Physics. PMID- 12780219 TI - Iterated function systems and dynamical systems. AB - We study the relationship between measures invariant for a piecewise expanding transformation tau of a compact metric space endowed with a underlying measure and measures invariant for an iterated function system T(tau), generated by inverse branches of tau. The main result says that the tau-invariant absolutely continuous measure &mgr; is also T(tau) invariant if and only if tau is absolutely continuously conjugated with a piecewise linear transformation. Measures of maximal entropy and general equilibrium states are also discussed. (c) 1995 American Institute of Physics. PMID- 12780220 TI - Complex dynamics and multistability in a damped harmonic oscillator with delayed negative feedback. AB - A center manifold reduction and numerical calculations are used to demonstrate the presence of limit cycles, two-tori, and multistability in the damped harmonic oscillator with delayed negative feedback. This model is the prototype of a mechanical system operating with delayed feedback. Complex dynamics are thus seen to arise in very plausible and commonly occurring mechanical and neuromechanical feedback systems. (c) 1995 American Institute of Physics. PMID- 12780221 TI - On the Riemannian description of chaotic instability in Hamiltonian dynamics. AB - In this work we investigate Hamiltonian chaos using elementary Riemannian geometry. This is possible because the trajectories of a standard Hamiltonian system (i.e., having a quadratic kinetic energy term) can be seen as geodesics of the configuration space manifold equipped with the standard Jacobi metric. The stability of the dynamics is tackled with the Jacobi-Levi-Civita equation (JLCE) for geodesic spread and is applied to the case of a two degrees of freedom Hamiltonian. A detailed comparison is made among the qualitative informations given by Poincare sections and the results of the geometric investigation. Complete agreement is found. The solutions of the JLCE are also in quantitative agreement with the solutions of the tangent dynamics equation. The configuration space manifold associated to the Hamiltonian studied here is everywhere of positive curvature. However, curvature is not constant and its fluctuations along the geodesics can yield parametric instability of the trajectories, thus chaos. This mechanism seems to be one of the most effective sources of chaotic instabilities in Hamiltonians of physical interest, and makes a major difference with Anosov flows, and, in general, with abstract geodesic flows of ergodic theory. (c) 1995 American Institute of Physics. PMID- 12780222 TI - From Hamiltonian chaos to Maxwell's Demon. AB - The problem of the existence of Maxwell's Demon (MD) is formulated for systems with dynamical chaos. Property of stickiness of individual trajectories, anomalous distribution of the Poincare recurrence time, and anomalous (non Gaussian) transport for a typical system with Hamiltonian chaos results in a possibility to design a situation equivalent to the MD operation. A numerical example demonstrates a possibility to set without expenditure of work a thermodynamically non-equilibrium state between two contacted domains of the phase space lasting for an arbitrarily long time. This result offers a new view of the Hamiltonian chaos and its role in the foundation of statistical mechanics. (c) 1995 American Institute of Physics. PMID- 12780223 TI - Chaos in thermal pulse combustion. AB - An experimental thermal pulse combustor and a differential equation model of this device are shown to exhibit chaotic behavior under certain conditions. Chaos arises in the model by means of a progression of period-doubling bifurcations that occur when operating parameters such as combustor wall temperature or air/fuel flow are adjusted to push the system toward flameout. Bifurcation sequences have not yet been reproduced experimentally, but similarities are demonstrated between the dynamic features of pressure fluctuations in the model and experiment. Correlation dimension, Kolmogorov entropy, and projections of reconstructed attractors using chaotic time series analysis are demonstrated to be useful in classifying dynamical behavior of the experimental combustor and for comparison of test data to the model results. Ways to improve the model are suggested. (c) 1995 American Institute of Physics. PMID- 12780224 TI - A simple model of chaotic advection and scattering. AB - In this work, we study a blinking vortex-uniform stream map. This map arises as an idealized, but essential, model of time-dependent convection past concentrated vorticity in a number of fluid systems. The map exhibits a rich variety of phenomena, yet it is simple enough so as to yield to extensive analytical investigation. The map's dynamics is dominated by the chaotic scattering of fluid particles near the vortex core. Studying the paths of fluid particles, it is seen that quantities such as residence time distributions and exit-vs-entry positions scale in self-similar fashions. A bifurcation is identified in which a saddle fixed point is created upstream at infinity. The homoclinic tangle formed by the transversely intersecting stable and unstable manifolds of this saddle is principally responsible for the observed self-similarity. Also, since the model is simple enough, various other properties are quantified analytically in terms of the circulation strength, stream velocity, and blinking period. These properties include: entire hierarchies of fixed points and periodic points, the parameter values at which these points undergo conservative period-doubling bifurcations, the structure of the unstable manifolds of the saddle fixed and periodic points, and the detailed structure of the resonance zones inside the vortex core region. A connection is made between a weakly dissipative version of our map and the Ikeda map from nonlinear optics. Finally, we discuss the essential ingredients that our model contains for studying how chaotic scattering induced by time-dependent flow past vortical structures produces enhanced diffusivities. (c) 1995 American Institute of Physics. PMID- 12780225 TI - On vortex entities of 2D turbulence in wavenumber space. AB - To connect vortices in physical space and scales in wavenumber space, spectral definitions for vortex size and momentum are introduced within the framework of a probabilistic method. At a late stage of 2D decaying turbulence, a simple solution is given for the vortex position and momentum probabilities. From the solution, an energy spectrum E(k) for self-similar vortices is constructed, which is in agreement with that observed in numerical simulations. (c) 1995 American Institute of Physics. PMID- 12780226 TI - Painleve' analysis of a variable coefficient Sine-Gordon equation. AB - In this paper we study a variable coefficient Sine-Gordon (vSG) equation given by theta(tt)-theta(xx)+F(x,t)sin theta=0 where F(x,t) is a real function. To establish if it may be integrable we have performed the standard test of Weiss, Tabor, and Carnevale (WTC). We have got that the (vSG) equation has the Painleve' property (Pp) if the function F(x,t) satisfies a well-defined nonlinear partial differential equation. We have found the general solution of this last equation and, consequently, the functions F(x,t) such that the (vSG) equation possesses the (Pp), are given by F(x,t)=F(1)(x+t)F(2)(x-t) where F(1)(x+t) and F(2)(x-t) are arbitrary functions. Using this last result we have obtained some particular solutions of the vSG equation. (c) 1995 American Institute of Physics. PMID- 12780227 TI - Master-slave synchronization from the point of view of global dynamics. AB - We present a mathematical framework for the theory of a synchronization phenomenon for dynamical systems discovered by Pecora and Carroll [Phys. Rev. Lett. 64, 821-824 (1990)]. From this perspective, we can synchronize, using a single coordinate, an open dense set of linear systems. We use our insights to synchronize nonlinear systems which were not previously recognized as being synchronizable. (c) 1995 American Institute of Physics. PMID- 12780228 TI - Wave trains in a model of gypsy moth population dynamics. AB - A recent model of gypsy moth [Lymantria dispar (Lepidoptera: Lymantriidae)] populations led to the observation of traveling waves in a one-dimensional spatial model. In this work, these waves are studied in more detail and their nature investigated. It was observed that when there are no spatial effects the model behaves chaotically under certain conditions. Under the same conditions, when diffusion is allowed, traveling waves develop. The biomass densities involved in the model, when examined at one point in the spatial domain, are found to correspond to a limit cycle lying on the surface of the chaotic attractor of the spatially homogeneous model. Also observed are wave trains that have modulating maxima, and which when examined at one point in the spatial domain show a quasiperiodic temporal behavior. This complex behavior is determined to be due to the interaction of the traveling wave and the chaotic background dynamics. (c) 1995 American Institute of Physics. PMID- 12780230 TI - Symmetries and regular behavior of Hamiltonian systems. AB - The behavior of the phase trajectories of the Hamilton equations is commonly classified as regular and chaotic. Regularity is usually related to the condition for complete integrability, i.e., a Hamiltonian system with n degrees of freedom has n independent integrals in involution. If at the same time the simultaneous integral manifolds are compact, the solutions of the Hamilton equations are quasiperiodic. In particular, the entropy of the Hamiltonian phase flow of a completely integrable system is zero. It is found that there is a broader class of Hamiltonian systems that do not show signs of chaotic behavior. These are systems that allow n commuting "Lagrangian" vector fields, i.e., the symplectic 2 form on each pair of such fields is zero. They include, in particular, Hamiltonian systems with multivalued integrals. (c) 1996 American Institute of Physics. PMID- 12780229 TI - Erratum: "Effect of a magnetic flux line on the quantum beats in the Henon-Heiles level density" [Chaos 5, 317 (1995)]. PMID- 12780231 TI - An averaging method for Hamiltonian systems, exponentially close to integrable ones. AB - Exponentially small separatrix splitting for a pendulum with rapidly oscillating suspension point and for the standard Chirikov map is studied by means of a new averaging method, which is a continuous version of the Neishtadt averaging procedure. An asymptotic formula for the rate of the separatrix splitting is obtained. (c) 1996 American Institute of Physics. PMID- 12780232 TI - Dynamical phenomena in systems with structurally unstable Poincare homoclinic orbits. AB - Recent results describing non-trivial dynamical phenomena in systems with homoclinic tangencies are represented. Such systems cover a large variety of dynamical models known from natural applications and it is established that so called quasiattractors of these systems may exhibit rather non-trivial features which are in a sharp distinction with that one could expect in analogy with hyperbolic or Lorenz-like attractors. For instance, the impossibility of giving a finite-parameter complete description of dynamics and bifurcations of the quasiattractors is shown. Besides, it is shown that the quasiattractors may simultaneously contain saddle periodic orbits with different numbers of positive Lyapunov exponents. If the dimension of a phase space is not too low (greater than four for flows and greater than three for maps), it is shown that such a quasiattractor may contain infinitely many coexisting strange attractors. (c) 1996 American Institute of Physics. PMID- 12780233 TI - Universality in the quasiperiodic route to chaos. AB - Numerous physical systems with two competing frequencies exhibit frequency locking and chaos associated with quasiperiodicity. In this paper we review certain universal aspects of the quasiperiodic route to chaos by making use of the standard circle map. Particular attention is paid to the golden mean and silver mean with a view to comparison with experimental work. (c) 1996 American Institute of Physics. PMID- 12780234 TI - A two-parameter study of the extent of chaos in a billiard system. AB - The billiard system of Benettin and Strelcyn [Phys. Rev. A 17, 773-785 (1978)] is generalized to a two-parameter family of different shapes. Its boundaries are composed of circular segments. The family includes the integrable limit of a circular boundary, convex boundaries of various shapes with mixed dynamics, stadiums, and a variety of nonconvex boundaries, partially with ergodic behavior. The extent of chaos has been measured in two ways: (i) in terms of phase space volume occupied by the main chaotic band; and (ii) in terms of the Lyapunov exponent of that same region. The results are represented as a kind of phase diagram of chaos. We observe complex regularities, related to the bifurcation scheme of the most prominent resonances. A detailed stability analysis of these resonances up to period six explains most of these features. The phenomenon of breathing chaos [Nonlinearity 3, 45-67 (1990)]-that is, the nonmonotonicity of the amount of chaos as a function of the parameters-observed earlier in a one parameter study of the gravitational wedge billiard, is part of the picture, giving support to the conjecture that this is a fairly common global scenario. (c) 1996 American Institute of Physics. PMID- 12780235 TI - The dripping faucet revisited. AB - High accuracy experimental results on the nonlinear dynamical behaviour of a dripping faucet are presented. The distribution functions for droplet sizes and drip intervals together with return maps are studied for various dripping rates. Increasing this control parameter, chaotic behaviour is obtained and discussed. (c) 1996 American Institute of Physics. PMID- 12780236 TI - Initial conditions of closed classical orbits from quantum spectra. AB - A method is presented for determining the initial conditions of classical orbits from the quantum spectra of the diamagnetic hydrogen atom. Each classical trajectory which is closed at the nucleus produces a sinusoidal fluctuation in the photoabsorption spectrum. The amplitude of each orbit's contribution appears in the Fourier transform of a spectrum computed at constant scaled energy. For a given initial state, closed-orbit theory gives the dependence of this recurrence amplitude on the initial angle of an orbit. By comparing the recurrence amplitudes for different initial states, the initial conditions of closed classical orbits are determined from quantum spectra. (c) 1996 American Institute of Physics. PMID- 12780237 TI - Changes in the adiabatic invariant and streamline chaos in confined incompressible Stokes flow. AB - The steady incompressible flow in a unit sphere introduced by Bajer and Moffatt [J. Fluid Mech. 212, 337 (1990)] is discussed. The velocity field of this flow differs by a small perturbation from an integrable field whose streamlines are almost all closed. The unperturbed flow has two stationary saddle points (poles of the sphere) and a two-dimensional separatrix passing through them. The entire interior of the unit sphere becomes the domain of streamline chaos for an arbitrarily small perturbation. This phenomenon is explained by the nonconservation of a certain adiabatic invariant that undergoes a jump when a streamline crosses a small neighborhood of the separatrix of the unperturbed flow. An asymptotic formula is obtained for the jump in the adiabatic invariant. The accumulation of such jumps in the course of repeated crossings of the separatrix results in the complete breaking of adiabatic invariance and streamline chaos. (c) 1996 American Institute of Physics. PMID- 12780238 TI - Catastrophic extinction, noise-stabilized turbulence and unpredictability of competition in a modified Volterra-Lotka model. AB - Spatial coexistence and competition among species is investigated through a modified Volterra-Lotka model which takes into account sexual breeding. This allows the population specific growth rate to depend on the population density. As a result of this modification the degeneracy inherent in the classical model is eliminated and qualitatively novel regimes are observed, as demonstrated by parametric analysis of the model. In the case where the corresponding parameters of competing species do not differ significantly the model can be reduced to a single Ginzburg-Landau type equation. The spatially distributed model is analyzed both in the absence and in the presence of noise mimicking inherent fluctuations in birth and death rates. It is shown that noise can qualitatively change the behavior of the system. Not only does it induce the formation of spatial patterns, but also switches on endless turbulent-like rearrangement of the system. When initially unpopulated habitat is occupied by competing species even a very low-intensity noise makes the final state of the system totally unpredictable and sensitive to any fluctuations. (c) 1996 American Institute of Physics. PMID- 12780239 TI - Nonlinear noise reduction for electrocardiograms. AB - The electrical activity of the heart usually shows dynamical behavior which is neither periodic nor deterministically chaotic: The interbeat intervals seem to contain a random component. Although long term predictions are thus impossible, good predictions can be made for times smaller than one heart cycle. This fact is used in order to suppress measurement errors by a local geometric projection method which was originally developed for chaotic signals. The result constitutes evidence that techniques of time series analysis based on chaos theory can be useful despite the fact that very few natural phenomena have been actually established to be deterministically chaotic. (c) 1996 American Institute of Physics. PMID- 12780240 TI - Musical variations from a chaotic mapping. AB - A chaotic mapping provides a technique for generating musical variations of an original work. This technique, based on the sensitivity of chaotic trajectories to initial conditions, produces changes in the pitch sequence of a piece. A sequence of musical pitches {p(i)}, i.e., any piece ranging from Bach (or earlier) to contemporary music, is paired with the x-components {x(i)} of a Lorenz chaotic trajectory. Each p(i) is marked on the x axis at the point designated by its x(i). In this way, the x axis becomes a pitch axis configured according to the notes of the original composition. Then, a second chaotic trajectory, whose initial condition differs from the first, is launched. Its x components trigger pitches on the pitch axis (via the mapping) that vary in sequence from the original work, thus creating a variation. There are virtually an unlimited number of variations possible, many appealing to expert and nonexpert alike. (c) 1996 American Institute of Physics. PMID- 12780241 TI - On noninvertible mappings of the plane: Eruptions. AB - In this paper we are concerned with the dynamics of noninvertible transformations of the plane. Three examples are explored and possibly a new bifurcation, or "eruption," is described. A fundamental role is played by the interactions of fixed points and singular curves. Other critical elements in the phase space include periodic points and an invariant line. The dynamics along the invariant line, in two of the examples, reduces to the one-dimensional Newton's method which is conjugate to a degree two rational map. We also determine, computationally, the characteristic exponents for all of the systems. An unexpected coincidence is that the parameter range where the invariant line becomes neutrally stable, as measured by a zero Lyapunov exponent, coincides with the merging of a periodic point with a point on a singular curve. (c) 1996 American Institute of Physics. PMID- 12780242 TI - Piecewise linear models for the quasiperiodic transition to chaos. AB - We formulate and study analytically and computationally two families of piecewise linear degree one circle maps. These families offer the rare advantage of being non-trivial but essentially solvable models for the phenomenon of mode locking and the quasiperiodic transition to chaos. For instance, for these families, we obtain complete solutions to several questions still largely unanswered for families of smooth circle maps. Our main results describe (1) the sets of maps in these families having some prescribed rotation interval; (2) the boundaries between zero and positive topological entropy and between zero length and non zero length rotation interval; and (3) the structure and bifurcations of the attractors in one of these families. We discuss the interpretation of these maps as low-order spline approximations to the classic "sine-circle" map and examine more generally the implications of our results for the case of smooth circle maps. We also mention a possible connection to recent experiments on models of a driven Josephson junction. (c) 1996 American Institute of Physics. PMID- 12780243 TI - Regular and chaotic motions in applied dynamics of a rigid body. AB - Periodic and regular motions, having a predictable functioning mode, play an important role in many problems of dynamics. The achievements of mathematics and mechanics (beginning with Poincare) have made it possible to establish that such motion modes, generally speaking, are local and form "islands" of regularity in a "chaotic sea" of essentially unpredictable trajectories. The development of computer techniques together with theoretical investigations makes it possible to study the global structure of the phase space of many problems having applied significance. A review of a number of such problems, considered by the authors in the past four or five years, is given in this paper. These include orientation and rotation problems of artificial and natural celestial bodies and the problem of controlling the motion of a locomotion robot. The structure of phase space is investigated for these problems. The phase trajectories of the motion are constructed by a numerical implementation of the Poincare point map method. Distinctions are made between regular (or resonance), quasiregular (or conditionally periodic), and chaotic trajectories. The evolution of the phase picture as the parameters are varied is investigated. A large number of "phase portraits" gives a notion of the arrangement and size of the stability islands in the "sea" of chaotic motions, about the appearance and disappearance of these islands as the parameters are varied, etc. (c) 1996 American Institute of Physics. PMID- 12780244 TI - Enlarged scaling ranges for the KS-entropy and the information dimension. AB - Numerical estimates of the Kolmogorov-Sinai entropy based on a finite amount of data decay towards zero in the relevant limits. Rewriting differences of block entropies as averages over decay rates, and ignoring all parts of the sample where these rates are uncomputable because of the lack of neighbours, yields improved entropy estimates. In the same way, the scaling range for estimates of the information dimension can be extended considerably. The improvement is demonstrated for experimental data. (c) 1996 American Institute of Physics. PMID- 12780245 TI - Semiclassical limit for a truncated Hamiltonian. AB - In the numerical calculation of the eigenenergies of a polynomial Hamiltonian, the majority of the levels depend on the cutoff of the basis used. By analyzing the finite Hamiltonian matrix as corresponding to a classical "Action Billiard" we are able to explain several features of the full spectrum using semiclassical periodic orbit theory. There are a large number of low-period orbits which interfere at the higher energies contained in the billiard. In this range the billiard becomes more regular than the untruncated Hamiltonian, as reflected by the Berry-Robnik level spacing distribution. (c) 1996 American Institute of Physics. PMID- 12780246 TI - Classical and quantum localization and delocalization in the Fermi accelerator, kicked rotor and two-sided kicked rotor models. AB - The phenomena of dynamical localization, both classical and quantum, are studied in the Fermi accelerator model. The model consists of two vertical oscillating walls and a ball bouncing between them. The classical localization boundary is calculated in the case of "sinusoidal velocity transfer" [A. J. Lichtenberg and M. A. Lieberman, Regular and Stochastic Motion (Springer-Verlag, Berlin, 1983)] on the basis of the analysis of resonances. In the case of the "sawtooth" wall velocity we show that the quantum localization is determined by the analytical properties of the canonical transformations to the action and angle coordinates of the unperturbed Hamiltonian, while the existence of the classical localization is determined by the number of continuous derivatives of the distance between the walls with respect to time. (c) 1996 American Institute of Physics. PMID- 12780247 TI - The quantized D-transformation. AB - We construct a new example of a quantum map, the quantized version of the D transformation, which is the natural extension to two dimensions of the tent map. The classical, quantum and semiclassical behavior is studied. We also exhibit some relationships between the quantum versions of the D-map and the parity projected baker's map. The method of construction allows a generalization to dissipative maps which includes the quantization of a horseshoe. (c) 1996 American Institute of Physics. PMID- 12780248 TI - Dynamical behavior of the multiplicative diffusion coupled map lattices. AB - We report a dynamical study of multiplicative diffusion coupled map lattices with the coupling between the elements only through the bifurcation parameter of the mapping function. We discuss the diffusive process of the lattice from an initially random distribution state to a homogeneous one as well as the stable range of the diffusive homogeneous attractor. For various coupling strengths we find that there are several types of spatiotemporal structures. In addition, the evolution of the lattice into chaos is studied. A largest Lyapunov exponent and a spatial correlation function have been used to characterize the dynamical behavior. (c) 1996 American Institute of Physics. PMID- 12780249 TI - Homoclinic connections and period doublings of a ship advancing in quartering waves. AB - The large-amplitude motions of a ship running in waves are analyzed with a mathematical model reduced to a system of eight coupled nonlinear ordinary differential equations. Bifurcation analysis in relation to the surf-riding condition, with control parameter the angle of the rudder, shows the existence of a region of oscillatory behavior, containing also a chaotic domain. This region ends with a homoclinic connection and a dangerous jump toward the overtaking-wave mode, which can incur ship capsize. Addition of linear control removes the chaotic domain while giving rise to new regions of oscillation. (c) 1996 American Institute of Physics. PMID- 12780250 TI - Pseudo-spectral methods and linear instabilities in reaction-diffusion fronts. AB - We explore the application of a pseudo-spectral Fourier method to a set of reaction-diffusion equations and compare it with a second-order finite difference method. The prototype cubic autocatalytic reaction-diffusion model as discussed by Gray and Scott [Chem. Eng. Sci. 42, 307 (1987)] with a nonequilibrium constraint is adopted. In a spatial resolution study we find that the phase speeds of one-dimensional finite amplitude waves converge more rapidly for the spectral method than for the finite difference method. Furthermore, in two dimensions the symmetry preserving properties of the spectral method are shown to be superior to those of the finite difference method. In studies of plane/axisymmetric nonlinear waves a symmetry breaking linear instability is shown to occur and is one possible route for the formation of patterns from infinitesimal perturbations to finite amplitude waves in this set of reaction diffusion equations. (c) 1996 American Institute of Physics. PMID- 12780251 TI - Chaos and intermittent bursting in a reaction-diffusion process. AB - Karhunen-Loeve decomposition is done on a chaotic spatio-temporal solution obtained from a nonlinear reaction-diffusion model of a chemical system simulating a chemical process in an open Couette-flow reactor. Using a Galerkin projection of the dominant Karhunen-Loeve modes back onto the nonlinear partial differential system, we obtain an ordinary differential equation model of the same process. Major features such as intermittent and chaotic bursting of the nonlinear process as well as the mechanism of transition to chaos are shown to exist in the low-dimensional model as well as the PDE model. From the low dimensional model the onset of intermittent bursts followed by small amplitude oscillations is shown to arise due to a sequence of saddle-node bifurcations. PMID- 12780252 TI - Collective patterns arising out of spatio-temporal chaos. AB - We present a simple mathematical model in which a time averaged pattern emerges out of spatio-temporal chaos as a result of the collective action of chaotic fluctuations. Our evolution equation possesses spatial translational symmetry under periodic boundary conditions. Thus the spatial inhomogeneity of the statistical state arises through spontaneous symmetry breaking. The transition from a state of homogeneous spatio-temporal chaos to one exhibiting spatial order is explained by introducing a collective viscosity which relates the averaged pattern with a correlation of the fluctuations. (c) 1996 American Institute of Physics. PMID- 12780253 TI - Statistical measures derived from the correlation integrals of physiological time series. AB - In this work correlation integrals are used for the analysis of various EEG signals from rabbits in resting states and under the influence of an anesthetic. The comparison with surrogate data reveals nonlinear dynamics in all of the time series. Our attempt to determine the correlation dimension D(2) by the modified algorithm of Theiler [Phys. Rev. A 34, 2427 (1986)] failed since no saturation is reached with increasing embedding dimension. The hypothesis of low-dimensional chaos turns out to be inconsistent with our results, but we can still distinguish, at least qualitatively, between different states of brain dynamics. A quantitative characterization of the time series is possible by defining correlation parameters P(a) derived from correlation integrals reflecting also autocorrelation of the signal. (c) 1996 American Institute of Physics. PMID- 12780254 TI - Erratum: "Steady vortices in plasmas and geophysical flows" [Chaos 4, 253 (1994)]. PMID- 12780255 TI - Overview: Synchronization and patterns in complex systems. AB - The theory of complex systems, such as neural assemblies or lattices of chaotic oscillators has generated many new problems including the synchronization or regularization of the cooperative behavior of systems consisting of chaotic elements, regular spatial patterns in "chaotic" lattices, and so on. A number of these problems were discussed at the International School in Nonlinear Science-95 (Nizhniy Novgorod, Russia). In this overview we try to formulate some of the most interesting problems that were discussed at that meeting. (c) 1996 American Institute of Physics. PMID- 12780256 TI - Images of synchronized chaos: Experiments with circuits. AB - Synchronization of oscillations underlies organized dynamical behavior of many physical, biological and other systems. Recent studies of the dynamics of coupled systems with complex behavior indicate that synchronization can occur not only in case of periodic oscillations, but also in regimes of chaotic oscillations. Using experimental observations of chaotic oscillations in coupled nonlinear circuits we discuss a few forms of cooperative behavior that are related to the regimes of synchronized chaos. This paper is prepared under the request of the editors of the special focus issue of Chaos and contains the materials for the lecture at the International School in Nonlinear Science, "Nonlinear Waves: Synchronization and Patterns," Nizhniy Novgorod, Russia, 1995. The main goal of the paper is to outline the collection of examples that illustrate the state of the art of chaos synchronization. (c) 1996 American Institute of Physics. PMID- 12780257 TI - Bifurcation analysis of nephron pressure and flow regulation. AB - One- and two-dimensional continuation techniques are applied to study the bifurcation structure of a model of renal flow and pressure control. Integrating the main physiological mechanisms by which the individual nephron regulates the incoming blood flow, the model describes the interaction between the tubuloglomerular feedback and the response of the afferent arteriole. It is shown how a Hopf bifurcation leads the system to perform self-sustained oscillations if the feedback gain becomes sufficiently strong, and how a further increase of this parameter produces a folded structure of overlapping period-doubling cascades. Similar phenomena arise in response to increasing blood pressure. The numerical analyses are supported by existing experimental results on anesthetized rats. (c) 1996 American Institute of Physics. PMID- 12780258 TI - Dynamics and kinematics of simple neural systems. AB - The dynamics of simple neural systems is of interest to both biologists and physicists. One of the possible roles of such systems is the production of rhythmic patterns, and their alterations (modification of behavior, processing of sensory information, adaptation, control). In this paper, the neural systems are considered as a subject of modeling by the dynamical systems approach. In particular, we analyze how a stable, ordinary behavior of a small neural system can be described by simple finite automata models, and how more complicated dynamical systems modeling can be used. The approach is illustrated by biological and numerical examples: experiments with and numerical simulations of the stomatogastric central pattern generators network of the California spiny lobster. (c) 1996 American Institute of Physics. PMID- 12780259 TI - An advanced investigation of interaction of allocated quasi-two-dimensional vortices. AB - This work should be regarded as a natural development of the investigations by Dolzhanskii, Krymov and Manin [Sov. Phys. Usp. 33, 495-520 (1990); J. Fluid Mech. 241, 705-722 (1992); Russ. J. Comput. Model. 1, 107-118 (1993)] of quasi-two dimensional (Q2D) flows in which the linear and weakly nonlinear stability theory based on the 2D hydrodynamic equations with the Rayleigh (Ekman) friction term imitating the influence of the bottom on the motion of upper fluid layers was corroborated with laboratory and observational data. The applicability of the Q2D approach to describe self-oscillating supercritical regimes was even more vague as Batchaev's experiments [Izv. AN SSSR Fiz. Atmos. Okeana 25, 434-439 (1989); Z. Prikl. Mech. Tech. Fiz., No. 4, 85-91 (1990)] on modeling the four vortex self oscillations in a thin fluid layer by the magnetohydrodynamics method (the so called hydrodynamical clock [Obukhov, Dolzhanskii and Batchaev, Topological Fluid Mechanics, Proceedings of the IUTAM Symposium, Cambridge, 13-18 August 1989 (Cambridge University Press, Cambridge, 1989), pp. 304-314]) did not find an appropriate theoretical explanation. To remove earlier uncontrolled effects the supplementary detailed measurements of the experimental flow characteristics were implemented, including the spatial spectral composition of the external vorticity sources and free surface 2D velocity fields. Satisfactory agreement is found between experimentally measured flow characteristics and the results of numerical simulations. The frequency of self-oscillations was found to be greatly susceptible to the spectral composition of the external vorticity sources and fluid layer thickness, which should be taken into account in designing laboratory experiments to simulate the natural Q2D processes observed in the ocean and atmosphere. Applicability conditions of the Q2D approach and the influence of geometrical parameters of vortices on their nonlinear interplay are also discussed. (c) 1996 American Institute of Physics. PMID- 12780260 TI - Self-organization of the large-scale planetary and plasma drift vortices. AB - This paper is a semi-review. A new understanding of the self-organization mechanism of solitary (i.e., long-lived and, in this sense, soliton-like) large scale vortices in geophysical fluid dynamics, as well as that of drift vortices in the magnetized plasma is discussed. This understanding differs from that described in a review paper by Nezlin [Chaos 4, 187-202 (1994)]. Earlier it was believed that formation of the solitary Rossby (and plasma drift) vortices was a result of equilibrium between wave dispersion and KdV-type nonlinearity. Under the influence of experimental data obtained by our team [M. V. Nezlin and E. N. Snezhkin, Rossby Vortices, Spiral Structures, Solitons (Springer-Verlag, Heidelberg, 1993)], it became obvious that the self-organization of the structures inevitably includes an essential effect of other nonlinearities; first, that presented by the Jacobian in the equations. (We replace the term "Rossby soliton" by the more exact one "the Rossby solitary vortex.") It must be noted from the very beginning that the term "self-organization" is used mainly in context with an explanation which factors (dispersion and nonlinearities of different kind) condition formation of the solitary (stable, long-lived) Rossby structures. Although, the experimental fact (see below) that the size of solitary vortices turns out to be close to the Rossby-Obukhov radius, independently of the size of the vortex local source, calls to mind the formation of an attractor. In essence, the Rossby solitary vortex self-organization process (although, only for the case of anticyclones) was described by Nycander and Sutyrin [Dyn. Atmos. Oceans 16, 473-498 (1992)]. Unfortunately, however, the authors did not use the term "self-organization." Our description, being in accord with Nycander and Sutyrin, relates not only to anticyclones, but also to anticyclones and cyclones. Second, a description of the experimental discovery of "anomalous" cyclonic anticyclonic asymmetry is given. Unlike "normal" asymmetry which manifests, in particular, in that the big vortices dominating in giant planet atmospheres (e.g., the Great Red Spot of Jupiter, the Brown Spot of Saturn, the Great Dark Spot of Neptune, et al.) are anticyclones, the asymmetry described manifests in that large-scale solitary vortices may be as cyclones only, not anticyclones. This phenomenon is observed in the presence of a rather strong and properly directed gradient in rotating shallow water depth. This type of asymmetry exists with drift vortices in magnetized plasma. Third, the physical difference between the planetary atmosphere and the laboratory model based on the shallow water layer in a rotating paraboloid is discussed (following, in principle, the Nycander-93 work). It is shown that laboratory modeling is adequate. Fourth, it is also shown that the most essential behavior of the vortices studied on the so called "beta-plane" of planets and in plasmas can be described by means of rather simplified and visual equations. These are the so-called generalized Charney Obukhov equation in fluid dynamics and its plasma counterpart, the generalized Hasegawa-Mima equation. Finally, nonlinearities are revealed, which condition properties of the geostrophic vortices under study on the "f-plane," i.e., in the polar regions of planets. &c PMID- 12780261 TI - Light induced annihilation and shift of spiral waves. AB - The light-induced collapse of a pair of spiral waves was studied in a chemically active medium based on the photosensitive Ru(bpy)(3)-catalyzed Belousov Zhabotinsky reaction. Spiral waves annihilate only if the light intensity is increased in proper phase relative to the spiral waves' rotation. Otherwise, the distance between spiral wave cores increases and the pair survives. Computer simulations reveal the mechanism which forces the spiral waves to collide and annihilate. It is based on the shift of a single spiral wave upon an instantaneous decrease of excitability of the medium. (c) 1996 American Institute of Physics. PMID- 12780262 TI - Drift mechanism caused by a nonlinear wave and the Cassini Division and Uranian rings formation. AB - The mechanism leading to the observed coexistence of gaps and narrow ringlets in the planetary rings is found. It is based upon the quasi-stationary radial drift of the matter under action of two forces in the disk plane: the Coriolis force and the Reynolds stresses. To an accuracy of the factor of 2 the first force coincides with the Lorentz force, therefore the radial drift in rings is similar to the gradient drift of plasma in the magnetic field. The second force is produced by the wave generated by the nearby satellite in the resonance position. In inertial systems, the second force alone causes a matter flow in its direction, called acoustic streaming. Since the radial drift is caused by nonlinear time-averaged force of high-frequency harmonic interactions in the wave, it exists in the wave propagation zone: from the birth place of the wave the resonance position, up to the reflection point of the wave, where its group velocity vanishes. Our estimations show that the size of the density wave propagation zone corresponding to the density wave which had been formerly generated the 2:1 orbital resonance with Mimas is consistent with the width of the Cassini Division. In our case the nature of the radial drift is such that first of all it clears out the farthest from the resonance position; later, the closer areas also get affected by the drift. The zone closest to the resonance position itself is the last to be involved in the process. The matter carried away by the drift is partially accumulated near the resonance position forming a narrow dense ringlet. (c) 1996 American Institute of Physics. PMID- 12780263 TI - Complex spatiotemporal convection patterns. AB - This paper reviews recent efforts to describe complex patterns in isotropic fluids (Rayleigh-Benard convection) as well as in anisotropic liquid crystals (electro-hydrodynamic convection) when driven away from equilibrium. A numerical scheme for solving the full hydrodynamic equations is presented that allows surprisingly well for a detailed comparison with experiments. The approach can also be useful for a systematic construction of models (order parameter equations). (c) 1996 American Institute of Physics. PMID- 12780264 TI - Nonlinear waves, chaos and patterns in microwave electronic devices. AB - We discuss some problems, concerning the application of nonlinear dynamics methods and ideas to vacuum microwave electronics. We consider such phenomena as solitons, deterministic chaos and pattern formation in different models of electron flows and devices. Our results reveal that microwave electronics is an interesting field of application of nonlinear dynamics. (c) 1996 American Institute of Physics. PMID- 12780265 TI - Formation and control of localized structures in nonlinear optical systems. AB - Diffractive effects in passive nonlinear optical resonators can lead to pattern forming instabilities. When the pattern (in our case, a regular hexagonal lattice of intensity peaks) coexists with the homogeneous solution, soliton-like intensity peaks in the transverse plane can be excited. These solutions have the characteristics of localized structures and are highly degenerate with respect to the peak location. By injecting narrow laser pulses, it is possible to turn on such peaks at desired locations and to turn them off selectively. The conditions to ensure independence among the peaks are described as well. These features suggest the possibility of encoding optical information in the structure of the field profile. (c) 1996 American Institute of Physics. PMID- 12780266 TI - Low-frequency dynamics of lasers. AB - An approach to nonlinear dynamics of multimode lasers is developed. It is based on the concept of two systems of eigenoscillations: optical modes and relaxation oscillations. The importance of a correct (not arbitrary) choice of a model is underlined. Characteristic features of two different rate equation models are formulated and compared. A method of selective perturbation on the system is described which makes it possible to study interrelations between optical modes and relaxation oscillations, and to control dynamical behavior of a laser. The possibility of using dynamical regularities for solving both applied and basic problems is illustrated in several examples. (c) 1996 American Institute of Physics. PMID- 12780267 TI - Wave collapse in plasmas and fluids. AB - This lecture is a review of recent results (obtained mainly by the author and his co-authors) in the wave collapse theory with applications to plasma physics, fluid dynamics and nonlinear optics as well. The main attention in the review is paid to the qualitative reasons of the wave collapse and to the exact methods based on the integral estimations. Both approaches are applied to both the nonlinear Schrodinger equation and the two-dimensional generalized Benjamin-Ono equation which describes self-focusing of low-frequency oscillations in the boundary layer. (c) 1996 American Institute of Physics. PMID- 12780268 TI - Quantum chaos. AB - In this paper we present an overview of important recent results in the study of a very controversial topic, the so-called quantum chaos. The theoretical and numerical results are compared with real laboratory experiments with special emphasis on the problem of ionization of hydrogen atoms in external microwave fields. (c) 1996 American Institute of Physics. PMID- 12780269 TI - Quantum chaos in gas. AB - A new approach to the description of gas kinetics is proposed. The approach is based on a fundamental presumption which states that binary collisions of gas atoms, moving in a chaotic way, can serve as "interior measurements." These "measurements" result in collapses of atomic wave functions. The ensuing decoherence is capable of transforming the wave function of gas atoms into a set of small-size wave packets. Their kinetics is considered in detail. Quantum effects linear in the Planck constant are taken into account. (c) 1996 American Institute of Physics. PMID- 12780270 TI - Nonlinear oscillating structures in the earthquake and seaquake dynamics. AB - The article deals with the problem of parametric excitation of oscillating structures during an earthquake and a seaquake associated with external forces. Tidal forces and their linear and nonlinear components are considered as possible causes of earthquakes. A seaquake is regarded as a typical large-scale structural disturbance of the water surface resulting from the ocean bottom earthquake. There are given results of original laboratory seaquake modeling where wave structures with hexagonal and square cells appeared. The received wave lattices were similar to Faraday ripples, but with the size of cells from 15 to 120 mm. These experiments provided parameters on transition from a wave structure to chaos. Comparison of laboratory experimental data with descriptions of full-scale seaquakes and parametric wave theory has confirmed the submitted interpretation of the phenomenon. (c) 1996 American Institute of Physics. PMID- 12780271 TI - Entropy estimation of symbol sequences. AB - We discuss algorithms for estimating the Shannon entropy h of finite symbol sequences with long range correlations. In particular, we consider algorithms which estimate h from the code lengths produced by some compression algorithm. Our interest is in describing their convergence with sequence length, assuming no limits for the space and time complexities of the compression algorithms. A scaling law is proposed for extrapolation from finite sample lengths. This is applied to sequences of dynamical systems in non-trivial chaotic regimes, a 1-D cellular automaton, and to written English texts. (c)1996 American Institute of Physics. PMID- 12780272 TI - Escape orbits for non-compact flat billiards. AB - It is proven that, under some conditions on f, the non-compact flat billiard Omega={(x,y) in R(0) (+)xR(0) (+); 0Phe (Y753F)/Y759F] and SH2 (Src homology 2) domains (R564A/R672A) in the context of Lyn-PLCgamma2, we found that Tyr753/Tyr759 were essential, whereas the PLCgamma2 SH2 domains did not have an important role in the transient activation of Lyn PLCgamma2 but may serve to stabilize an activated form in sustained activation. PMID- 12780342 TI - A fat-enriched, glucose-enriched diet markedly attenuates adiponectin mRNA levels in rat epididymal adipose tissue. AB - Adiponectin levels are decreased in subjects with obesity, diabetes and coronary artery disease. In the present study, we have investigated whether the decrease in the levels and mRNA expression of adiponectin is due to obesity or to the diet itself. Wistar rats were either fed standard laboratory chow throughout (controls) or given a fat-enriched, glucose-enriched diet (diet-fed) for 2 days or 16 weeks. After 2 days of diet feeding, total body weight, fat pad masses and the plasma levels of glucose, insulin and leptin were all comparable between the two groups, while plasma NEFA (non-esterified fatty acid) and triacylglycerol levels were increased in the diet-fed animals (P<0.01 for both). There was a marked (P<0.01) decrease in plasma adiponectin levels. After 16 weeks of diet feeding, diet-fed rats had significantly higher body weight, fat pad mass and plasma levels of leptin, adiponectin, NEFA and triacylglycerol (P<0.001 for all) compared with chow-fed controls, whereas plasma levels of glucose and insulin were similar in the two groups. After 2 days of diet feeding, there were no significant changes in Ob mRNA levels in epididymal fat, whereas there was a marked decrease in adiponectin mRNA levels. After 16 weeks of diet feeding, rats had significantly increased levels of Ob mRNA, but decreased adiponectin mRNA levels, in epididymal fat compared with the chow-fed group (P<0.001 for both). These findings suggest that obesity per se is not a factor in the decreased adiponectin levels observed in obese subjects. We propose that the lipid profile of the plasma and/or the constituents of the diet consumed by rats may contribute to adiponectin levels more than obesity per se. PMID- 12780344 TI - Decreased HLA (human leucocyte antigen)-DR expression on peripheral blood monocytes predicts the development of organ failure in patients with acute pancreatitis. AB - Immune suppression plays an important role in the pathogenesis of acute pancreatitis. Monocyte expression of HLA (human leucocyte antigen)-DR, a cellular marker of immune suppression, was determined in relation to the development of organ dysfunction in patients with acute pancreatitis. A total of 310 consecutive patients with acute pancreatitis, admitted to a university hospital within 72 h of pain onset, were studied; 194 (63%) had mild disease (group I), 87 (28%) had severe disease without organ dysfunction (group II), and 29 (9%) had severe disease with organ dysfunction (group III). HLA-DR expression, defined both as the proportion of monocytes that were HLA-DR-positive and as monocyte HLA-DR fluorescence intensity, was determined at admission, using whole-blood flow cytometry. Of the patients in group III, 13 (45%) developed organ dysfunction within 24 h of admission. The proportion of HLA-DR-positive monocytes and monocyte HLA-DR density were both related to the severity of pancreatitis (P<0.001 for linear trend). In predicting organ dysfunction, the sensitivity, specificity and positive-likelihood ratio for the proportion of HLA-DR-positive monocytes were 83% [95% CI (confidence interval) 64-94%], 72% (67-77%) and 3.0 respectively, and for monocyte HLA-DR density the respective values were 69% (49 85%), 84% (79-88%) and 4.3. In conclusion, monocyte HLA-DR expression predicts the development of organ dysfunction that occurs early in patients with acute pancreatitis. PMID- 12780343 TI - Regulation of hypoxia-inducible factor-1alpha by cyclical mechanical stretch in rat vascular smooth muscle cells. AB - Vascular smooth muscle cells (VSMCs) are exposed to hormonal and mechanical stress in vivo. Hormonal factors have been shown to affect hypoxia-inducible factor-1alpha (HIF-1alpha). How mechanical stress affects the regulation of HIF 1alpha in VSMCs has not been reported previously, and therefore we sought to investigate the regulation of HIF-1alpha by cyclical mechanical stretch in cultured rat VSMCs. Rat VSMCs grown on a flexible membrane base were stretched by vacuum to 20% of the maximum elongation at 60 cycles/min. The levels of HIF 1alpha protein began to increase as early as 2 h after stretch was applied and reached a maximum of 2.8-fold over the control by 4 h. Real-time PCR showed that the levels of HIF-1alpha mRNA increased 2.1-fold after cyclical stretch for 4 h. Cyclical mechanical stretch also increased the immunohistochemical labelling of HIF-1alpha in VSMCs after cyclical stretch for 4 h. The phosphorylation of p42/p44 mitogen-activated protein kinase (MAP kinase) increased after stretch and this was inhibited by the MAP kinase kinase inhibitors PD98059 and U0126. PD98059 and U0126 also blocked HIF-1alpha gene expression induced by cyclical stretch. In conclusion, cyclical mechanical stretch activates the gene expression of HIF 1alpha in cultured VSMCs and this mechanical effect is possibly mediated by the p42/p44 MAP kinase kinase pathway. PMID- 12780345 TI - Effects of two lipid-lowering, carotenoid-controlled diets on the oxidative modification of low-density lipoproteins in free-living humans. AB - This study compares the effects of two lipid-lowering diets [a diet enriched in MUFAs (monounsaturated fatty acids) and a HCLF (high-carbohydrate/low-fat) diet] with a controlled carotenoid content on risk factors for coronary heart disease, including in vitro copper-induced LDL (low-density lipoprotein) oxidation and serum lipid levels. A randomized crossover dietary intervention study, with two diets each consumed for 14-16 days, was conducted in 18 women and 13 men aged 20 70 years, recruited via personal contacts and advertisements in newspapers. Both diets (MUFA-enriched diet and HCLF diet) contained the same basic foods and had a controlled carotenoid content, high in lycopene. The in vitro copper-induced oxidation of isolated LDL showed a longer lag phase (mean difference 7.4 min in women and 7.34 min in men) after the MUFA-enriched diet compared with the HCLF diet. Serum total cholesterol, LDL cholesterol and carotenoid levels were similar after the two diets. Serum triacylglycerol levels were significantly lower and those of HDL (high-density lipoprotein) cholesterol were significantly higher at the end of the MUFA-enriched diet compared with the HCLF diet. It is concluded that the significantly longer lag phase for oxidation of LDL, the higher HDL cholesterol level and the lower triacylglycerol level in subjects following a carotenoid-controlled, MUFA-enriched diet may decrease the risk of coronary heart disease. PMID- 12780346 TI - Differential expression of Flk-1 and Flt-1 in rat skeletal muscle in response to chronic ischaemia: favourable effect of muscle activity. AB - To determine the involvement of vascular endothelial growth factor (VEGF) and its receptors Flk-1 and Flt-1 in capillary growth in ischaemic skeletal muscle, extensor digitorum longus muscles from hindlimbs of Sprague-Dawley rats were studied at 1, 2 and 5 week intervals after iliac artery ligation. Muscle VEGF protein levels (as determined by Western-blot analysis) increased only after 2 (60%) and 5 (80%) weeks, with more capillaries positively immunostained for VEGF than in control muscles. Ischaemia-induced angiogenesis was gradual, with capillary proliferation at 1 and 2 weeks and capillary:fibre ratio increased 20% after 5 weeks. This was associated with an initial doubling of Flk-1 protein after 1 week that declined below control levels by 5 weeks, whereas Flt-1 expression was elevated more than 40% at all time points. During more sustained ischaemia (femoral ligation 3 weeks after iliac ligation), VEGF protein level at 5 weeks was even higher, but Flt-1 and Flk-1 were unchanged from control levels and no capillary growth occurred. Intermittent electrical stimulation (10 Hz, 7x15 min/day) of these ischaemic muscles between weeks 3-5 did not elevate VEGF further, but increased Flk-1 by 32%, decreased Flt-1 by 71%, and led to significant capillary growth. These results demonstrate that during chronic muscle ischaemia Flk-1 and Flt-1 are regulated differentially and that electrical stimulation of ischaemic muscles can promote angiogenesis via Flk-1 up regulation. Even when ischaemic muscle VEGF levels are high, capillary growth appears to be dependent on the presence of Flk-1. PMID- 12780347 TI - Response of yeast to the regulated expression of proteins in the Bcl-2 family. AB - The mechanisms by which pro-apoptotic members of the Bcl-2 family of proteins promote the release of mitochondrial factors like cytochrome c, subsequently activating the apoptotic cascade, or by which anti-apoptotic family members block this release, are still not understood. When expressed in yeast, Bcl-2 family members act directly upon conserved mitochondrial components that correspond to their apoptotic substrates in mammalian cells. Here we describe a system in which the levels of representative pro- and anti-apoptotic members of the Bcl-2 family can be regulated independently in yeast. Using this system, we have focused on the action of the anti-apoptotic family member Bcl-x(L), and have defined the quantitative relationships that underlie the antagonistic action of this protein on the lethal consequences of expression of the pro-apoptotic family member Bax. This system has also allowed us to demonstrate biochemically that Bcl-x(L) has two actions at the level of the mitochondrion. Bcl-x(L) is able to inhibit the stable integration of Bax into mitochondrial membranes, as well as hinder the action of Bax that does become stably integrated into these membranes. Taken together, our results suggest that both the functional and biochemical actions of Bcl-x(L) may be based on the ability of this molecule to disrupt the interaction of Bax with a resident mitochondrial target that is required for Bax action. Finally, we confirm that VDAC (voltage-dependent anion channel) is not required for the functional responses observed following the expression of either pro- or anti-apoptotic members of the Bcl-2 family. PMID- 12780348 TI - XB51 isoforms mediate Alzheimer's beta-amyloid peptide production by X11L (X11 like protein)-dependent and -independent mechanisms. AB - XB51 (derived from X11-like binding protein of clone number 51) was isolated by yeast two-hybrid cDNA screening using the N-terminal domain of X11L (X11-like protein) as a bait. X11L is a neuron-specific adaptor protein that is known to down-regulate APP (beta-amyloid precursor protein) metabolism by associating with the cytoplasmic domain of APP, but the detailed mechanisms are still unknown. Thus the X11L-associated protein XB51 is believed to regulate APP metabolism by modifying X11L function through its interaction with X11L. Here we report that the hXB51 (human XB51 ) gene can yield two transcripts, one with exon 9 spliced out (resulting in the hXB51beta isoform) and the other containing exon 9 (yielding the hXB51alpha isoform). hXB51alpha binds to X11L to form a tripartite complex composed of hXB51alpha, X11L and APP. Complex-formation results in blocking X11L's suppression of Abeta (beta-amyloid) generation from APP. hXB51beta associates with X11L and inhibits its interaction with APP. However, hXB51beta suppresses Abeta generation and secretion in an X11L-independent manner. Thus the hXB51 isoforms regulate Abeta generation differently, either enhancing it by modifying the association of X11L with APP or suppressing it in an X11L-independent manner. These observations advance our understanding of the molecular mechanisms regulating intracellular Abeta production and the pathogenesis of Alzheimer's disease. PMID- 12780349 TI - Characterization of human torsinA and its dystonia-associated mutant form. AB - Deletion of a single glutamate in torsinA correlates with early-onset dystonia, the most severe form of a neurological disorder characterized by uncontrollable muscle contractions. TorsinA is targeted to the ER (endoplasmic reticulum) in eukaryotic cells. We investigated the processing and membrane association of torsinA and the dystonia-associated Glu-deletion mutant (torsinAdeltaE). We found that the signal sequence of torsinA (residues 1-20 from the 40 amino-acid long N terminal hydrophobic region) is cleaved in Drosophila S2 cells, as shown by the N terminal sequencing after partial protein purification. TorsinA is not secreted from S2 cells. Consistently, sodium carbonate extraction and Triton X-114 treatment showed that torsinA is associated with the ER membrane in CHO (Chinese hamster ovary) cells. In contrast, a variant of torsinA that contains the native signal sequence without the hydrophobic region Ile24-Pro40 does not associate with the membranes in CHO cells, and a truncated torsinA without the 40 N terminal amino acids is secreted in the S2 culture. Thus the 20-amino-acid-long hydrophobic segment in torsinA, which remains at the N-terminus after signal peptide cleavage, is responsible for the membrane anchoring of torsinA. TorsinAdeltaE showed similar cleavage of the 20 N-terminal amino acids and membrane association properties similar to wild-type torsinA but, unlike the wild type, torsinAdeltaE was not secreted in the S2 culture even after deletion of the membrane-anchoring segment. This indicates that the dystonia-associated mutation produces a structurally distinct, possibly misfolded, form of torsinA, which cannot be properly processed in the secretory pathway of eukaryotic cells. PMID- 12780351 TI - The evolving contribution of gambling research to addiction theory. PMID- 12780352 TI - Polydrug use and overdose: overthrowing old myths. PMID- 12780350 TI - Regulation of alternative splicing of Gtf2ird1 and its impact on slow muscle promoter activity. AB - A human MusTRD (muscle TFII-I repeat domain (RD)-containing protein) isoform was originally identified in a yeast one-hybrid screen as a protein that binds the slow fibre-specific enhancer of the muscle gene troponin I slow [O'Mahoney, Guven, Lin, Joya, Robinson, Wade and Hardeman (1998) Mol. Cell. Biol. 18, 6641 6652]. MusTRD shares homology with the general transcription factor TFII-I by the presence of diagnostic I-RDs [Roy (2001) Gene 274, 1-13]. The human gene encoding MusTRD, GTF2IRD1 ( WBSCR11 / GTF3 ), was subsequently located on chromosome 7q11.23, a region deleted in the neurodegenerative disease, Williams-Beuren Syndrome [Osborne, Campbell, Daradich, Scherer, Tsui, Franke, Peoples, Francke, Voit, Kramer et al. (1999) Genomics 57, 279-284; Franke, Peoples and Francke (1999) Cytogenet. Cell. Genet. 86, 296-304; Tassabehji, Carette, Wilmot, Donnai, Read and Metcalfe (1999) Eur. J. Hum. Genet. 7, 737-747]. The haploinsufficiency of MusTRD has been implicated in the myopathic aspect of this disease, which manifests itself in symptoms such as lowered resistance to fatigue, kyphoscoliosis, an abnormal gait and joint contractures [Tassabehji, Carette, Wilmot, Donnai, Read and Metcalfe (1999) Eur. J. Hum. Genet. 7, 737-747]. Here, we report the identification of 11 isoforms of MusTRD in mouse skeletal muscles. These isoforms were isolated from a mouse skeletal muscle cDNA library and reverse transcription-PCR on RNA from various adult and embryonic muscles. The variability in these isoforms arises from alternative splicing of a combination of four cassettes and two mutually exclusive exons, all in the 3' region of the primary transcript of Gtf2ird1, the homologous mouse gene. The expression of some of these isoforms is differentially regulated spatially, suggesting individual regulation of the expression of these isoforms. Co-transfection studies in C2C12 muscle cell cultures reveal that isoforms differentially regulate muscle fibre type-specific promoters. This indicates that the presence of different domains of MusTRD influences the activity exerted by this molecule on multiple promoters active in skeletal muscle. PMID- 12780353 TI - Rethinking alcohol, tobacco and other drug control. PMID- 12780354 TI - Legalization of illicit drugs: two sides to the coin. PMID- 12780355 TI - Public health and drugs: why is policy so persistently irrational? PMID- 12780356 TI - The inherent limits of regulation. PMID- 12780357 TI - Sitting on the beach: comments on Room. PMID- 12780358 TI - Learning from tobacco: bans on commercial availability are not unthinkable. PMID- 12780360 TI - Are scratchcards addictive? The prevalence of pathological scratchcard gambling among adult scratchcard buyers in the Netherlands. AB - AIMS: To determine the prevalence of regular, potential problematic and pathological scratchcard gambling (PSG) 5 years after the introduction of scratchcards in the Netherlands. DESIGN AND PARTICIPANTS: A non-proportional stratified random sample of 12,222 scratchcard buyers was approached. Regular scratchcard buyers (n = 3342) were asked to fill out the South Oaks Gambling Screen (SOGS). Those with a SOGS score of 3 or more (n = 340) were interviewed with the gambling section of the DSM-IV Diagnostic Interview Schedule (DIS-T). Weighted data were used to obtain unbiased prevalence estimates. FINDINGS: The estimated prevalence of regular and potential problematic scratchcard gambling were 28.4% and 2.68%, respectively. Only 0.24% met DSM-IV criteria for PSG. Of those, only 0.09% were addicted uniquely to scratchcards. The remaining 0.15% were also addicted to other games of chance. Demographic and gambling characteristics of these 'combined' PSG (young men, mainly slot-machine players) resembled characteristics of pathological gamblers in general. In contrast to these 'combined' PSG, 'unique' PSG were mainly women between 25 and 34 years who spent relatively small amounts of money on scratchcards (equivalent to one scratchcard a day). CONCLUSION: Scratchcards have a very low addiction potential among adults in the Netherlands. Given the specific characteristics of the unique PSG and the relatively small amount of money they spent, the appropriateness of DSM criteria for this particular form of gambling can be questioned. PMID- 12780361 TI - Video lottery: winning expectancies and arousal. AB - AIMS: This study investigates the effects of video lottery players' expectancies of winning on physiological and subjective arousal. DESIGN: Participants were assigned randomly to one of two experimental conditions: high and low winning expectancies. SETTING: Participants played 100 video lottery games in a laboratory setting while physiological measures were recorded. Level of risk taking was controlled. PARTICIPANTS: Participants were 34 occasional or regular video lottery players. They were assigned randomly into two groups of 17, with nine men and eight women in each group. INTERVENTION: The low-expectancy group played for fun, therefore expecting to win worthless credits, while the high expectancy group played for real money. MEASUREMENTS: Players' experience, demographic variables and subjective arousal were assessed. Severity of problem gambling was measured with the South Oaks Gambling Screen. In order to measure arousal, the average heart rate was recorded across eight periods. FINDINGS: Participants exposed to high as compared to low expectations experienced faster heart rate prior to and during the gambling session. According to self-reports, it is the expectancy of winning money that is exciting, not playing the game. CONCLUSIONS: Regardless of the level of risk-taking, expectancy of winning is a cognitive factor influencing levels of arousal. When playing for fun, gambling becomes significantly less stimulating than when playing for money. PMID- 12780362 TI - Opiates, cocaine and alcohol combinations in accidental drug overdose deaths in New York City, 1990-98. AB - AIMS: Accidental drug overdose contributes substantially to mortality among drug users. Multi-drug use has been documented as a key risk factor in overdose and overdose mortality in several studies. This study investigated the contribution of multiple drug combinations to overdose mortality trends. DESIGN: We collected data on all overdose deaths in New York City between 1990 and 1998 using records from the Office of the Chief Medical Examiner (OCME). We standardized yearly overdose death rates by age, sex and race to the 1990 census population for NYC to enable comparability between years relevant to this analysis. FINDINGS: Opiates, cocaine and alcohol were the three drugs most commonly attributed as the cause of accidental overdose death by the OCME, accounting for 97.6% of all deaths; 57.8% of those deaths were attributed to two or more of these three drugs in combination. Accidental overdose deaths increased in 1990-93 and subsequently declined slightly in 1993-98. Changes in the rate of multi-drug combination deaths accounted for most of the change in overdose death rates, whereas single drug overdose death rates remained relatively stable. Trends in accidental overdose death rates within gender and racial/ethnic strata varied by drug combination suggesting different patterns of multi-drug use among different subpopulations. CONCLUSIONS: These data suggest that interventions to prevent accidental overdose mortality should address the use of drugs such as heroin, cocaine and alcohol in combination. PMID- 12780363 TI - Reasons for selecting an initial route of heroin administration and for subsequent transitions during a severe HIV epidemic. AB - AIM: To identify the most important reasons for selecting a particular route of heroin administration and for subsequent transitions during a period of epidemic HIV transmission. To study temporal trends in these reasons. DESIGN: Cross sectional survey. PARTICIPANTS: Nine hundred heroin users in three Spanish cities: 305 in Seville, 297 in Madrid and 298 in Barcelona. MEASUREMENTS: A separate analysis was made of the reasons for five types of behaviour: (a) selecting injection as the initial usual route of heroin administration (URHA); (b) changing the URHA to injection; (c) never having injected drugs; (d) selecting the smoked or sniffed route as the initial URHA; and (e) changing the URHA to a non-injected route. Subjects were invited to evaluate the importance of each reason included in a closed list. Spontaneously self-perceived reasons were also explored in an open-ended question for each of the five types of behaviour studied. FINDINGS: The primary reason selected for each type of behaviour was: (a) pressure of the social environment; (b) belief that injection is a more efficient route than smoking or sniffing heroin; (c) concern about health consequences (especially fears of HIV and overdose), and fear of blood or of sticking a needle into one's veins; (d), pressure of the social environment and (e) concern about health consequences and vein problems. For women, having a sexual partner who injected heroin played a decisive role in initiating or changing to injection. Few people spontaneously mentioned market conditions for purchasing heroin as an important reason for any behaviour, nor did many mention risk of overdose as reasons for (c) or (d). CONCLUSIONS: These findings should be considered when designing interventions aimed at preventing initiation of injecting or facilitating the transition to non-injected routes. PMID- 12780364 TI - Marginality among older injectors in today's illicit drug culture: assessing the impact of ageing. AB - AIMS: As conceptualized by Robert Park in 1928, the 'marginal man' occupies simultaneously an intermediate point between two distinct cultures that embody some level of contradictory normative expectations, behaviors and beliefs. This paper explores the influence of ageing and social change on the creation of marginality among injection drug users over age 50 who have been chronic heroin users since their youth. DESIGN: Our data are drawn from in-depth interviews with 40 (28 male and 12 female) not-in-drug-treatment, active injecting drug users between the ages of 18 and 68. SETTING: Our sample was recruited using street outreach as part of a larger, community-based study conducted from a converted storefront in a high crime, economically depressed neighborhood on the west side of Chicago. FINDINGS: As they aged, and under the pressures of social change, the older users whom we studied moved from center positioning in the illicit drug culture of their youth to the margins of a differing drug culture of today where they participate largely unseen by others. Nostalgic for the 'Old School' mores of the past, and unable to transcend or assimilate fully into the cultural practices and norms of the 'New', they respond to their predicament by embracing 'poise' in the face of loneliness, stress and fear of victimization. CONCLUSIONS: Our respondents' experiences draw attention to the need to refine the field's theoretical and practical understandings of what it means to be marginal and its consequences for older drug users and people in general. The results also point to what happens when society changes but some of its members do not. PMID- 12780365 TI - Factors associated with volatile solvent use among junior high school students in Kanto, Japan. AB - AIMS: To estimate the relative association between life-time volatile solvent use and risk factors for usage. DESIGN: Cross-sectional anonymous questionnaire survey. SETTING: Junior high schools in Kanto, Japan. PARTICIPANTS: Junior high school students (n = 7744). MEASUREMENTS: Data on life-time and past-year solvent use, demographic variables, urbanization, regularity of waking patterns, school life, family life, peer relationships, prior alcohol and cigarette use and knowledge on harmful effects of solvent use. FINDINGS: Uni- and multivariate logistic regression analyses were conducted to estimate crude and adjusted odds ratios for each index. The primary findings were (1) 'smoking cigarettes nearly every day' (adjusted OR = 9.88, 95% Cl = 3.74, 26.12) and peer pressure measured by 'been tempted to use solvents' (adjusted OR = 9.53, 95% Cl = 4.84, 18.74) demonstrated the highest adjusted odds ratios; (2) being male (adjusted OR = 2.56, 95% Cl = 1.37, 4.76), seeing school life as 'not at all enjoyable' (adjusted OR = 2.69, 95% Cl = 1.03, 7.01) and family environment as 'neither good nor bad' (adjusted OR = 2.15, 95% Cl = 1.19, 3.88) also showed significant association; (3) life-time alcohol use did not show a significant association in the multivariate model (adjusted OR = 0.80, 95% Cl = 0.30, 2.12); and (4) solvent use appeared independent of knowledge regarding its effects ('death by acute intoxication', 'psychotic symptoms', 'amotivational syndrome', 'flashbacks' adjusted ORs all non-significant). CONCLUSION: Alcohol use may not function as a gateway to solvent use in Japan. The reasons may be culture-bound. A longitudinal study is required to test this hypothesis. PMID- 12780366 TI - Maternal drug use and length of neonatal unit stay. AB - AIMS: Infants with neonatal abstinence syndrome (NAS) may require a prolonged neonatal unit admission, which has implications for both their families and bed occupancy. The aim of this study was to test the hypothesis that the duration of neonatal unit stay would be influenced by the type of maternal drug use and particularly prolonged for the infants whose mothers had taken methadone with other substances. DESIGN: The medical records of infants born at term who were admitted consecutively to a neonatal unit because of NAS were reviewed. Data were collected regarding antenatal and neonatal factors likely to affect neonatal stay. Comparisons were then made between three groups of infants: those whose mothers took methadone alone, methadone plus other drugs or non-methadone opioids. SETTING: Level three neonatal intensive care unit. PARTICIPANTS: Forty one infants with a median gestational age of 39 (range 37-42) weeks. FINDINGS: The 41 infants had a median duration of admission of 30 (range 3-68) days. Thirty six of the infants required treatment for NAS; their median duration of treatment was 29 (range 6-68) days. The duration of stay and requirement for treatment were greater in the infants exposed to methadone and other drugs compared to those exposed to non-methadone opiods only (P = 0.0212, P = 0.0343, respectively). The duration of stay without requirement for treatment was also longest in the methadone plus other drugs group (P = 0.0117). CONCLUSIONS: Prolonged treatment and neonatal unit stay are influenced by the type of maternal drug abused. PMID- 12780367 TI - Characteristics of non-respondents in a US national longitudinal survey on drinking and intimate partner violence. AB - AIMS: To identify non-respondents' characteristics in a longitudinal household survey of intimate partner violence and drinking among United States couples. DESIGN: Multistage probability sampling procedure with over-samples from black and Hispanic populations. A total of 1635 couples 18 years of age or older were interviewed at baseline (1995) for a response rate of 85%. Five years later, 1392 respondents were re-interviewed, with a response rate of 72%. MEASUREMENTS: Sociodemographic variables, drinking volume and pattern, drug use, intimate partner violence, exposure to parental violence and childhood physical abuse. FINDINGS: Among men, those who are younger, those who did not graduate from high school, those who are unemployed and those who are drinkers are more likely to be among the non-respondents. Among women, those who are 40-49 years of age are two times more likely than those aged 50 years or older to be among non-respondents. Also, women who reported experiencing abuse during their childhood are less likely to be among non-respondents compared with those who were not victimized. Gender-specific regression models accounted for only 5% of the variance in the response status variable, even after the inclusion of a comprehensive range of predictors in the model. CONCLUSIONS: A low proportion of the variance in survey participation was explained by the variables included in the analysis, suggesting that sociodemographic, drinking-related and violence-related variables might not be the main factors determining participation in the survey. PMID- 12780368 TI - Prevalence of alcohol use disorders and associated factors: a population-based study using AUDIT in southern Brazil. AB - AIMS: To assess the prevalence of potential alcohol use disorders and associated factors using the Alcohol Use Disorders Identification Test (AUDIT). DESIGN: Cross-sectional study. SETTING: A town in southern Brazil. PARTICIPANTS: A representative sample of 1260 people aged 15 and over. MEASUREMENTS: Demographic, socioeconomic, smoking habit and mental health data were collected. Logistic regression was used in the multivariate analysis, and odds ratios (ORs) and 95% confidence intervals (CIs) were calculated. FINDINGS: Overall prevalence of alcohol use disorder was 7.9%, with 14.5% prevalence among men and 2.4% among women. The risk of alcohol misuse increased across social class (P linear trend = 0.03) and compared with the highest classes (A and B), groups C through E had ORs of 1.48, 1.51 and 2.36, respectively. Males had an OR of 6.89 (CI 3.61-13.16) compared with women. A linear trend was found (P = 0.001) between smoking categories, and smokers (OR 3.27; CI 1.91-5.58) and ex-smokers (OR 1.30; CI 0.56 2.98) were at higher risk than non-smokers. Those with minor psychiatric disorders had a 2.48 OR (CI 1.35-4.56) of presenting a positive test. CONCLUSIONS: The AUDIT detected a high prevalence of potential alcohol use disorders in the population sampled. Those identified are potential targets for preventive measures implemented through health policies. PMID- 12780369 TI - Probabilities of alcohol high-risk drinking, abuse or dependence estimated on grounds of tobacco smoking and nicotine dependence. AB - AIMS: To estimate probabilities of alcohol high-risk drinking, alcohol abuse and alcohol dependence on grounds of smoking-behaviour related variables and single nicotine dependence criteria. DESIGN: Cross-sectional population-based study. SETTING: Adult population of a region in north Germany. PARTICIPANTS: Cigarette smokers (n = 2437) among a random sample of 4075 females and males aged 18-64, drawn in 1996. MEASUREMENT: Smoking, nicotine dependence according to the Diagnostic and Statistical Manual of Psychiatric Disorders (DSM-IV) and the Fagerstrom Test for Nicotine Dependence (FTND); increasing alcohol-related harm (ARH): high-risk drinking, DSM-IV alcohol abuse, remitted and current alcohol dependence diagnosed by the Composite International Diagnostic Interview (CIDI). FINDINGS: Having smoked 30 cigarettes or more per day, onset of smoking at the age of 17 or younger, nicotine dependence and single nicotine dependence criteria revealed odds ratios higher than 4.0 for alcohol dependence. For alcohol dependence, a logistic regression model showed an increased odds ratios for male gender, smoking for 25 years or more, no attempt to quit or cut down, continuation of smoking despite problems, craving for nicotine, withdrawal experience 1 day or longer, smoking first cigarette in the morning 5 minutes or less after waking. The probability of increasing ARH was more likely in males, smokers for 25 years or more, no attempt to quit or cut down, continuation of smoking despite problems and smoking first cigarette in the morning 5 minutes or less after waking. CONCLUSIONS: Gender and single nicotine dependence criteria show particularly high probabilities of alcohol dependence and increasing ARH. Interventions need to take these connections into account. PMID- 12780370 TI - Family structure and adolescent cigarette smoking in eleven European countries. AB - AIMS: This study seeks to establish (1) if different types of non-traditional family structures are related equally to adolescent cigarette smoking; (2) if each type of family structure is related equally to adolescent smoking in different countries and (3) if differences in such patterns can be explained by the prevalence of such family structures in each country. DESIGN: Self-reported cigarette smoking among 33 978 students in Cyprus, France, Hungary, Iceland, Ireland, Lithuania, Malta, Slovak Republic, Slovenia, Sweden and the United Kingdom is analysed with multi-level hierarchical regression models. FINDINGS: Adolescents living with both biological parents smoke less than those living with single mothers, who in turn smoke less than those living with single fathers, mothers-stepfathers, or with neither biological parent. Living with fathers stepmothers is associated with less smoking than living with single fathers, mother-stepfathers, or with neither biological parent, but does not differ from living with both biological parents or single mothers. The effects of living with single mothers, single fathers, or with neither biological parent are stronger in countries where such family types are less common. Differences in the strength of effects between countries become non-significant once the prevalence of each family type has been taken into account. CONCLUSIONS: Adolescents living with both biological parents smoke less than their counterparts in most other family types, and adolescents living with single mothers or fathers-stepmothers smoke less than those living in other non-traditional family structures. The strength of this pattern varies inversely with the prevalence of such households in each country. PMID- 12780371 TI - Eye movements to smoking-related pictures in smokers: relationship between attentional biases and implicit and explicit measures of stimulus valence. AB - AIMS: To investigate biases in overt orienting of attention to smoking-related cues in cigarette smokers, and to examine the relationship between measures of visual orienting and the affective and motivational valence of smoking cues. DESIGN: Smokers and non-smokers took part in a single session in which their attentional and evaluative responses to smoking-related and matched control pictures were recorded. PARTICIPANTS: Twenty smokers and 25 non-smokers. MEASUREMENTS: Direction and duration of gaze was measured while participants completed a visual probe task. Subjective and cognitive-experimental measures of the motivational and affective valence of the stimuli were recorded. FINDINGS: Smokers, but not non-smokers, maintained their gaze for longer on smoking-related pictures than control pictures. They were also faster to detect probes that replaced smoking-related than control pictures, consistent with an attentional bias for smoking-related cues. Furthermore, smokers showed greater preferences for smoking-related than control pictures, compared with non-smokers, on both the subjective (explicit) and cognitive-experimental (implicit) indices of stimulus valence. Within smokers, longer initial fixations of gaze on smoking-related pictures were associated with a bias to rate the smoking pictures more positively, with greater approach tendencies for smoking pictures on the cognitive-experimental task, and with a greater urge to smoke. CONCLUSIONS: These results demonstrate that smokers show biased attentional orientating to smoking cues, which is related to craving and the affective and motivational valence of the stimuli. PMID- 12780379 TI - Current initiatives in drug abuse research in Spain. PMID- 12780372 TI - The effectiveness of personalized smoking cessation strategies for callers to a Quitline service. AB - AIM: To assess the effectiveness of a program of computer-generated tailored advice for callers to a telephone helpline, and to assess whether it enhanced a series of callback telephone counselling sessions in aiding smoking cessation. DESIGN: Randomized controlled trial comparing: (1) untailored self-help materials; (2) computer-generated tailored advice only, and (3) computer generated tailored advice plus callback telephone counselling. Assessment surveys were conducted at baseline, 3, 6 and 12 months. SETTING: Victoria, Australia. PARTICIPANTS: A total of 1578 smokers who called the Quitline service and agreed to participate. MEASUREMENTS: Smoking status at follow-up; duration of cessation, if quit; use of nicotine replacement therapy; and extent of participation in the callback service. FINDINGS: At the 3-month follow-up, significantly more (chi2(2) = 16.9; P < 0.001) participants in the computer-generated tailored advice plus telephone counselling condition were not smoking (21%) than in either the computer-generated advice only (12%) or the control condition (12%). Proportions reporting not smoking at the 12-month follow-up were 26%, 23% and 22%, respectively (NS) for point prevalence, and for 9 months sustained abstinence; 8.2, 6.0, and 5.0 (NS). In the telephone counselling group, those receiving callbacks were more likely than those who did not to have sustained abstinence at 12 months (10.2 compared with 4.0, P < 0.05). Logistic regression on 3-month data showed significant independent effects on cessation of telephone counselling and use of NRT, but not of computer-generated tailored advice. CONCLUSION: Computer generated tailored advice did not enhance telephone counselling, nor have any independent effect on cessation. This may be due to poor timing of the computer generated tailored advice and poor integration of the two modes of advice. PMID- 12780373 TI - Sex differences in the effects of stressful life events on changes in smoking status. AB - AIMS: Stressful life events known to be associated with substance use were examined to determine if there were sex-specific responses to stress resulting in changes in smoking status. PARTICIPANTS AND MEASUREMENTS: A community-based sample of ever smokers from the Americans' Changing Lives study (n = 1512, 45% female based on sample weights) was used to examine the interactive effects of sex and stressful life events on the likelihood of two outcomes; relapse among former smokers and failure to quit among current smokers. Logistic regression procedures were used to calculate odds ratios. Factors known to be associated with smoking status (e.g. depression, self-esteem, social support) were assessed as control variables. FINDINGS: In the sample of former smokers (n = 729) interpersonal loss events were associated with continued abstinence, whereas change of residence and adverse financial events were associated with increased occurrence of relapse. Women were more likely than men to relapse in response to a financial event. In the sample of current smokers (n = 783), financial events were associated with continued smoking, whereas health events were associated with increased likelihood of quitting. Women were more likely than men to continue smoking in the presence of an adverse financial event and less likely than men to quit in response to an adverse health event. CONCLUSIONS: Overall, stressful life events appear to have a greater deleterious effect on continued abstinence and the ability to quit smoking for women when compared to men. In particular, health and financial events are important risk factors for women and tobacco use. PMID- 12780380 TI - Emergence of a Pan-European College of Addictions. PMID- 12780384 TI - Age, cell signalling and cardioprotection. AB - For many years investigators have been researching methods of preconditioning the myocardium against ischaemia-induced damage; however, a majority of this research has been carried out in young animals and cells. Normal ageing is accompanied by changes in the human myocardium that decrease its capacity to tolerate and respond to various forms of stress. Also, the likelihood of experiencing an ischaemic stress and other cardiovascular complications increases as an individual ages; therefore, an aged population would benefit most from cardioprotective treatments. Methods currently known to provide cardioprotection (or preconditioning) include exercise, heat stress, oxidative stress, brief ischaemia, stretch and certain pharmacological interventions. It is unclear whether the aged myocardium can adapt to a preconditioning stimulus; however, many researchers have observed age-related alterations in the expression and activation of proteins key to the cardioprotective process. These proteins include heat shock protein 70 (HSP70), nitric oxide synthase (NOS), the sodium hydrogen exchanger (NHE), and the mitogen-activated protein (MAP) kinases c-Jun N terminal Kinase (JNK), extracellular signal-regulated kinase (ERK), and p38. Therefore, the purpose of the current review will be to outline the current knowledge of these cardioprotective agents in an aged myocardium. Interactions among the cardioprotective agents outlined herein suggest that age-related changes in the myocardium will need to be better understood before cardioprotective interventions that have been proved effective in young animals can be applied to an aged human population. PMID- 12780383 TI - Dose-dependent effects of acute exercise on PKC levels in rat heart: is PKC the heart's prophylactic? AB - Epidemiological studies have demonstrated that chronic exercise is cardioprotective, and recent evidence from our laboratory suggests a key role for protein kinase C (PKC)-dependent pathways, at least in part, as a cellular basis for this response. However, the dose-response relationship linking exercise volume and the time course of isoform-specific PKC activation are poorly understood. AIM: The purpose of this investigation was to determine the effects of acute exercise of varying durations on PKC subcellular distribution and phosphorylation in the rat left ventricle. METHODS: Adult (5 months) male Fischer 344 more rats were subjected to a single bout (OB) or 7 days (SB) of treadmill running (n = 6/group; 23 m min-1, 20 min), and compared with sedentary controls (SED; n = 8). Hearts were isolated immediately after [early window (EW); n = 3/group] or 24 h after the last exercise bout [late window (LW); n = 3/group] in OB and SD, respectively. Total PKC and subcellular distribution for the alpha, delta, epsilon, betaI, and betaII isoforms, as well as phosphorylated (phospho-) PKC epsilon (pSer729), PKC alpha (pSer657) and PKCdelta (pThr507) levels were assessed by western blotting. Protein kinase C epsilon and PKC alpha mRNA levels were assessed by real time polymerase chain reaction. RESULTS: Following OB, PKCbetaI protein levels were reduced, while total phospho-PKC epsilon (pSer729), PKC alpha (pSer657) and PKC delta (pThr507) levels were increased during EW (P < 0.05). Interestingly, total PKC delta (31%) and membrane-associated PKC alpha (24%) levels decreased from EW to LW (P < 0.05). In contrast, SB yielded chronic increases in total PKC epsilon (80.5%) levels and PKC delta (20.0%) levels (P < 0.03), with reversal of effects on phospho-PKC epsilon (Ser729), phospho-PKC alpha (Ser657) and phospho-PKC delta (Thr507) levels observed with OB. Reductions in total phospho-PKC alpha (Ser657) persisted at SB (26.1%; P < 0.02). Interestingly, mRNA levels for PKC epsilon were significantly increased following SB while PKC alpha mRNA levels were reduced, respectively. CONCLUSION: These data suggest that divergent patterns of PKC activation occur following OB and SB at both the transcriptional and translational levels. That similar patterns of PKC translocation are observed in experimental models of ischaemic preconditioning and genetic PKC manipulation provide evidence for a dose-dependent cardioprotective phenotype induced by physical activity. PMID- 12780385 TI - Coronary smooth muscle adaptation to exercise: does it play a role in cardioprotection? AB - Substantial evidence exists supporting the role of chronic exercise in reducing the incidence and severity of coronary vascular disease. Physical inactivity is an independent risk factor for coronary heart disease suggesting that the cardioprotective effect of exercise is due, in part, to an intrinsic adaptation within the coronary vasculature. Surprisingly, a paucity of information exists regarding the intrinsic cellular changes within the coronary vasculature associated with exercise training and even less is known regarding the effect of physical activity on long-term phenotypic modulation of coronary smooth muscle (CSM). The purpose of this symposium is to provide a concise update on the current knowledge regarding CSM adaptation to exercise training and the potential for these adaptations to contribute to exercise-induced cardioprotection. The potential role of CSM in exercise-induced cardioprotection will be approached from two perspectives. First, endurance exercise training effects on the regulation of coronary vasomotor tone via changes in CSM calcium regulation will be reviewed, i.e. short-term functional adaptation. Secondly, we will discuss potential long-term consequences of this altered calcium regulation, i.e. exercise-induced phenotypic modulation of CSM. We propose that exercise training alters CSM intracellular calcium regulation to reduce Ca2+-dependent activation of the contractile apparatus and Ca2+-dependent gene transcription and increase activation of sarcolemmal potassium channels. The overall effect is to increase the gain of the vasomotor system and maintain a stable, contractile CSM phenotype. PMID- 12780386 TI - Lowering of interstitial fluid pressure in rat trachea after substance P alone and in combination with calcitonin gene-related peptide. AB - Neurogenic inflammation is mediated following a release of sensory neuropeptides including calcitonin gene-related peptide (CGRP) and substance P (SP). The release of peptides can be mediated chemically with capsaicin, or electrically by stimulation of the vagal nerve, both inducing vasodilation, plasma protein extravasation and lowering of interstitial fluid pressure (Pif) which will contribute to the enhancement of oedema formation. AIM: Lowering of Pif has previously been demonstrated following intravenous (i.v.) treatment with CGRP, but it was not possible to demonstrate that SP had this effect under the same condition. METHODS: Micropuncture measurements of Pif in the submucosa, without opening of the trachea, was conducted on rats anaesthetized with pentobarbital sodium (50 mg kg-1) and cardiac arrest was induced with i.v. KCl. RESULTS: Pif in vehicle-treated animal averaged -1.7 +/- 0.4 (SD) mmHg (n = 9). Intravenous injection of SP induced significant lowering of Pif compared with control, both at low dose (0.47 nmol kg-1 body weight) with 1 min distribution time (P < 0.007, -4.2 +/- 2.3 mmHg) and at high dose with seconds of distribution time (P < 0.03, 4.2 +/- 1.6 mmHg). The same response was observed after treatment with SP co injected with CGRP. CONCLUSIONS: Substance P alone or in combination with CGRP is able to induce a rapid lowering of Pif showing that this peptide is a potent agent in increasing the hydrostatic driving pressure initially transporting fluid into the tissue during an acute inflammatory reaction. PMID- 12780387 TI - Ischaemic preconditioning alters the energy metabolism and protects the ischaemic myocardium in a stepwise fashion. AB - AIMS AND METHODS: Recently it was suggested that ischaemic preconditioning (IP) protects the myocardium in a graded pattern as assessed by myocardial infarct size estimation. Using tissue biopsies we investigated the impact of the proposed graded pattern of protection on myocardial energy state in an open-chest porcine model of IP with either one (1xIP) or four (4xIP) episodes of preconditioning. Furthermore, we evaluated the relationship between interstitial energy-related metabolite levels obtained by the microdialysis technique and the degree of subsequent ischaemic insult. RESULTS: During the long ischaemia the difference between pre-ischaemic and post-ischaemic total adenylate pools and the sum of adenylate breakdown products (adenosine, inosine and hypoxanthine) as well as tissue lactate levels appeared as follows: non-IP > 1xIP > 4xIP (P < 0.05). Moreover interstitial peak levels of lactate, hypoxanthine and taurine displayed a graded pattern analogous to the development of ischaemic damage, where non-IP > 1xIP > 4xIP. CONCLUSIONS: We present for the first time concordant energy metabolic and morphometric data in support of IP being a stepwise phenomenon for protection of the ischaemic myocardium. Furthermore, IP resulted in proportionally higher levels of hypoxanthine (relative to inosine) in the ischaemic myocardium, suggesting a different handling of adenine nucleotide breakdown products in the IP myocardium. PMID- 12780388 TI - Epidermal growth factor and insulin short-term increase hPepT1-mediated glycylsarcosine uptake in Caco-2 cells. AB - AIMS: Little is known about the physiological regulation of the human intestinal di/tri-peptide transporter, hPepT1. In the present study we evaluated the effects of epidermal growth factor (EGF) and insulin on hPepT1-mediated dipeptide uptake in the intestinal cell line Caco-2. METHODS: Caco-2 cells were grown on filters for 23-27 days. Apical dipeptide uptake was measured using [14C]glycylsarcosine([14C]Gly-Sar). HPepT1 mRNA levels were investigated using RT PCR, cytosolic pH was determined using the pH-sensitive fluorescent probe BCECF. RESULTS: Basolateral application of EGF increased [14C]Gly-Sar uptake with an ED50 value of 0.77 +/- 0.25 ng mL-1 (n = 3-6) and a maximal stimulation of 33 +/- 2% (n = 3-6). Insulin stimulated [14C]Gly-Sar uptake with an ED50 value of 3.5 +/ 2.0 ng mL-1 (n = 3-6) and a maximal stimulation of approximately 18% (n = 3-6). Gly-Sar uptake followed simple Michaelis-Menten kinetics. Km in control cells was 0.98 +/- 0.11 mM (n = 8) and Vmax was 1.86 +/- 0.07 nmol cm-2 min-1 (n = 8). In monolayers treated with 200 ng mL-1 of EGF, Km was 1.11 +/- 0.05 mM (n = 5) and Vmax was 2.79 +/- 0.05 nmol cm-2 min-1 (n = 5). In monolayers treated with 50 ng mL-1 insulin, Km was 1.03 +/- 0.08 mM and Vmax was 2.19 +/- 0.06 nmol cm-2 min-1 (n = 5). Kinetic data thus indicates an increase in the number of active transporters, following stimulation. The incrased Gly-Sar uptake was not accompanied by changes in hPepT1 mRNA, nor by measurable changes in cytosolic pH. CONCLUSIONS: Short-term stimulation with EGF and insulin caused an increase in hPepT1-mediated uptake of Gly-Sar in Caco-2 cell monolayers, which could not be accounted for by changes in hPepT1 mRNA or proton-motive driving force. PMID- 12780389 TI - Oxygen dependency of the adrenergic Na/H exchange in rainbow trout erythrocytes is diminished by a hydroxyl radical scavenger. AB - AIM: Potassium transport via the potassium chloride cotransporter in rainbow trout erythrocytes is increased by high oxygen tension. It appears that the effect of oxygen is mediated by reactive oxygen species, especially hydroxyl radicals. In contrast, the activity of adrenergically stimulated sodium proton exchange decreases with increasing oxygen tension. As available data suggest that the two transporters are regulated reciprocally, the present study was undertaken to evaluate, if hydroxyl radicals may inhibit sodium transport via the adrenergically stimulated sodium proton exchanger. METHODS: The effects of the hydroxyl radical scavenger, 2 mm mercaptopropionyl glycine (MPG), on the activity of the adrenergically activated sodium proton exchange in rainbow trout erythrocytes were examined by measuring unidirectional sodium flux, using radioactive isotope, and cellular water content. RESULTS: The activity of the sodium proton exchange increased with decreasing oxygen tension after adrenergic stimulation. When MPG was present during incubation, there was no statistically significant effect of oxygen tension on the adrenergically stimulated sodium proton exchange, whereby the activity of the transporter at atmospheric oxygen tension was markedly higher in the presence than in the absence of MPG. In the absence of adrenergic stimulation, MPG did not influence the transporter activity significantly at any oxygen tension. CONCLUSION: The data suggest that hydroxyl radicals are involved in the inhibition of the adrenergically stimulated sodium proton exchange at elevated oxygen tensions. PMID- 12780390 TI - Strength, workload, anaerobic intensity and the immune response to resistance exercise in women. AB - AIM: The mechanism linking exercise intensity to the magnitude of the immune response is not completely understood. The purpose of this investigation was to determine whether the immune response to resistance exercise was associated with (1) changes in workload or (2) anaerobic exercise intensity. METHODS: Previously untrained women underwent 6 months of resistance training for lower and upper body (TOTAL, n = 34) or for upper body alone (UPPER, n = 30). Lymphocyte subsets [T (CD3+), CD4+, CD8+, NK and B], functional markers (CD45RA+ and CD45RO+), and mitogen (phytohemagglutinin-M, concanavalin A and pokeweed mitogen) and superantigen (staphylococcus a. cowans)-stimulated proliferation were measured from blood samples collected pre- and post-exercise for a squat resistance exercise consisting of six sets of 10 repetitions at 75% of one repetition maximum. This protocol was performed before (T0) and after 3 (T3) and 6 months (T6) of training. RESULTS: Lymphocyte recruitment to the circulation and proliferation following resistance exercise did not differ between training groups at any time, although the TOTAL group performed at a higher workload as training progressed. With respect to anaerobic intensity, exercise-induced increases in NK, CD4+, CD8+ and B lymphocyte concentrations were 42 (P = 0.07), 76 (P < 0.05), 72 (P < 0.05) and 242% (P < 0.01) greater in women in the highest compared with the lowest post-exercise lactate quartiles. Lymphocyte proliferation did not differ between lactate quartiles. CONCLUSIONS: Anaerobic intensity, rather than increased strength and workload, is associated with the number of lymphocytes recruited to the circulation, but not T and B cell proliferation responses. PMID- 12780391 TI - Interaction of fibre type, potentiation and fatigue in human knee extensor muscles. AB - AIM: To examine the effect of fibre type on potentiation and fatigue. METHODS: Young men (n = 4 per group) with a predominance of type I [61.4 +/- 6.9% (SD), group I (GI)] or type II [71.8 +/- 9.2%, group II (GII)] fibres in vastus lateralis, performed a fatigue protocol of sixteen 5-s maximal voluntary isometric contractions (MVCs) of the right knee extensors. Maximal twitches and corresponding muscle action potentials (M-waves) were evoked before the first MVC, during the 3-s rest period after each MVC and at intervals during the 5-min recovery period after the last MVC. RESULTS: Group II [49.3 +/- 2.6% (SE)] had a greater decrease in MVC force than GI (22.8 +/- 6.2%) during the fatigue protocol. Group II (126.4 +/- 13.6%) showed greater twitch force potentiation early in the fatigue protocol than GI (38.2 +/- 2.3%), but greater depression at the end (33.7 +/- 13.7% vs.17.4 +/- 3.4%). Twitch time-to-peak torque (TPT) and half relaxation time (HRT) initially decreased but then increased as the fatigue protocol progressed; GII had a greater increase in HRT. During a 5-min recovery period twitch force increased above the prefatigue level and remained so until the end of the recovery period; the pattern was similar in GI and GII. Twitch TPT and HRT remained elevated during recovery. M-wave area increased throughout the fatigue protocol and the first part of recovery before returning to baseline values in GII, whereas there were no significant changes in GI. The interaction between potentiation and fatigue was amplified in GII early in the fatigue protocol with concurrently greater twitch and M-wave potentiation, and greater MVC force decrease and HRT increase. Late in the protocol, GII had a greater decrease in twitch and MVC force combined with greater M-wave potentiation. CONCLUSION: It is concluded that fibre type distribution influences potentiation and fatigue of the twitch, and potentiation of the M-wave during fatiguing exercise. PMID- 12780392 TI - Muscle performance following fatigue induced by isotonic and quasi-isometric contractions of rat extensor digitorum longus and soleus muscles in vitro. AB - AIM: To study the effect of contraction mode on fatigue development. METHODS: Muscle fatigue was induced by isotonic and quasi-isometric contractions in rat soleus (SOL) and extensor digitorum longus (EDL) muscles, using identical stimulation protocol (60 Hz, 400 ms s-1) for 100 s in SOL and 60 s in EDL. Fatigue was quantified as the decline in peak values of shortening, shortening velocity, relaxation and work during the isotonic contractions, and, correspondingly, of force, rate of force development, relaxation and work during the quasi-isometric contractions. Maximal test contractions (60 Hz, 1.5 s) performed before and after fatigue were analysed for decline in force development (Fmax), rate of force development (dF/dtmax) and relaxation (-dF/dtmax). RESULTS: Fmax declined to significantly lower values after isotonic than after quasi isometric fatiguing contractions (fatigued in percentage of unfatigued): 58.5 +/- 6.4% vs. 64.4 +/- 7.0% in SOL, and 30.4 +/- 4.1% vs. 33.3 +/- 3.6% in EDL, respectively. The same pattern was seen for dF/dtmax which decreased to: 46.3 +/- 9.9% vs. 52.3 +/- 8.5% in SOL, and 19.1 +/- 4.3% vs. 22.3 +/- 3.2% in EDL after isotonic and quasi-isometric contractions, respectively. Similarly, when comparing fatigue development during the two contraction modes, the respective fatigue variables decreased more rapidly and to lower levels during isotonic vs. quasi-isometric contractions. During maximal test contractions, the dynamic fatigue variables (+/-dF/dtmax) declined to significantly lower levels than Fmax. CONCLUSIONS: Fatigue development was significantly larger during isotonic vs. quasi-isometric contractions. The use of force as the only experimental fatigue variable may underestimate the functional impairment of fatigued muscle, neglecting the fatigue effect on time and length dimensions. PMID- 12780394 TI - Acta ophthalmologica and the net. PMID- 12780395 TI - Ocular complications and visual outcome in juvenile chronic arthritis: a 25-year follow-up study. AB - OBJECTIVE: Assessment of longterm ophthalmic outcome in juvenile chronic arthritis (JCA) with emphasis on visual acuity and identification of disease related parameters associated with rheumatic eye affection. MATERIAL: Sixty-five adults (52 females, 13 males) with a history of or still active JCA were assessed an average of 26.7 years after disease onset. METHODS: Cross-sectional, longterm, follow-up study including complete ophthalmological and rheumatological examinations. RESULTS: Evidence of ocular complications was found in 13 subjects (20%): 10 had had recurrent iritis, two band keratopathy, and one diplopia due to rheumatoid eye muscle involvement. The three single eyes with poor sight (<0.1) were all in the complication subgroup. Binocularly, all but one patient scored at least 6/9 as best corrected visual acuity; one, however, had severe glaucoma impaired visual fields. The refractive range was -8.12 to +6.0 D. There were no cases of keratoconjunctivitis sicca. Iritis was associated with early disease onset (p = 0.02), longer disease duration (p = 0.02) and a positive antinuclear antibodies (ANA) (p = 0.02). CONCLUSION: Ophthalmic complications were recorded in 20% of the study group, primarily in ANA positive patients with pauciarticular onset JCA. In addition to the two patients with severely affected bilateral visual status (3%), one of the 65 patients had acquired unilateral blindness. Half of the complications were late in that they were first recorded after the age of 16 years. Otherwise, the findings were in keeping with other studies of visual prognosis using non-selected series. PMID- 12780396 TI - Visual prognosis in immunocompetent patients with herpes zoster ophthalmicus. AB - PURPOSE: To determine the normal spectrum of ocular complications and associated visual outcome in patients with herpes zoster ophthalmicus. METHODS: This prospective observational cohort study included 73 immunocompetent adults with herpes zoster ophthalmicus, referred by their general practitioners within 7 days of skin rash onset. The follow-up period was 6 months. All patients received a 7 14-day course of systemic aciclovir treatment combined with longterm application of a lubricating ophthalmic ointment as long as the corneal epithelium was affected. Topical corticosteroids were strictly avoided in the acute phase of ocular disease. Acquired visual loss scores at 1, 2 and 6 months were based on best corrected visual acuity (BCVA) level and evaluation of the ophthalmological history and findings. RESULTS: Ophthalmic herpes zoster led to a variety of transient inflammatory reactions within the anterior eye segment of the involved side in 46 patients (63%), but did not seriously compromise their ultimate visual outcome. Mild to moderate visual loss, with corrected VA between 0.3 and 0.8, was found in 17 patients at 1 month (23%), in 10 patients at 2 months (14%) and in seven patients at 6 months follow-up (10%). None of the patients developed visual loss with a corrected VA of less than 0.3. CONCLUSION: Functional vision was retained in all ophthalmic zoster patients referred to the ophthalmologist in the acute phase of the disease by vigorous antiviral treatment and adequate prevention of corneal exposure. PMID- 12780397 TI - Type 1 diabetes patients with severe non-proliferative retinopathy may benefit from panretinal photocoagulation. AB - PURPOSE: To examine whether panretinal photocoagulation for severe non proliferative retinopathy in type 1 diabetes patients could halt the progression of retinopathy with subsequent vitreous haemorrhages and visual impairment. METHODS: During a 10-year follow-up study period of 344 type 1 diabetes patients, 81 subjects went through panretinal photocoagulation. Forty patients were treated for severe non-proliferative retinopathy (age at onset of diabetes 14 +/- 8 years, diabetes duration 18 + 10 years) and 41 for proliferative retinopathy (age at onset 15 +/- 10 years, diabetes duration 22 + 13 years). One randomly selected eye per patient forms the basis for the study. Metabolic control, systolic and diastolic blood pressure, serum creatinine and urinary albumin levels were measured and analysed yearly during the follow-up period. RESULTS: A total of 35% (14/40) of eyes treated for severe non-proliferative retinopathy developed neovascularizations during a mean time of 2.9 +/- 1.5 years. Vitreous haemorrhages were more frequent in eyes with proliferative retinopathy at treatment than in eyes with severe non-proliferative retinopathy (12/41 versus 2/40; p = 0.007). The number of vitrectomies due to vitreous haemorrhages in eyes treated for severe non-proliferative retinopathy tended to be lower (1/40 versus 6/41; p = 0.052). Before photocoagulation, visual acuity (VA) was similar in eyes with severe non-proliferative retinopathy and in those with proliferative retinopathy (1.0, 0.4-1.0 versus 1.0, 0.1-1.0; median and range). Visual impairment and blindness tended to develop more often in eyes treated for proliferative retinopathy compared to those treated for severe non-proliferative retinopathy (10/40 versus 4/40; p = 0.056). Eyes with neovascularizations at follow-up were more often visually impaired (VA < 0.5) than eyes without neovascularizations (15/55 versus 1/26; p = 0.016). CONCLUSION: In type 1 diabetes, panretinal photocoagulation may be beneficial even at the severe non proliferative retinopathy stage in terms of preventing vitreous haemorrhage, subsequent vitrectomy and visual impairment. PMID- 12780398 TI - Topical medication interferes with face-down positioning after macular hole surgery. AB - PURPOSE: To investigate whether different procedures of topical medication interfere with postoperative face-down positioning and thereby influence the anatomical success rate of macular hole surgery. METHODS: A total of 39 consecutive eyes with macular holes were operated with vitrectomy and fluid-gas exchange, followed by face-down positioning for 1 week. Postoperatively, the procedures for topical medication differed between two consecutive groups of patients. In the first group (16 patients), a conventional postoperative regimen of topical antibiotics and steroids was administered as eye drops six times daily. In the second group (23 patients), topical medication was administered as ointments once daily, while the patients kept their heads straight forwards and looked slightly downwards. RESULTS: The macular hole closed successfully in 10 (62.5%) of the 16 eyes in the eye drop group, and in 21 (91.3%) of the 23 eyes in the ointment group (p < 0.05). CONCLUSION: The present study indicates that frequent postoperative administration of eye drops reduces the success rate of macular hole surgery. To prevent patients from looking upwards during the instillation of topical antibiotics and steroids, the use of ointments is recommended. PMID- 12780399 TI - A conservative blepharoplasty may be a means of alleviating dry eye symptoms. AB - PURPOSE: Patients with dermatochalasis often complain of dry eye symptoms. In a prospective study, the influence of upper eyelid blepharoplasty on dry eye symptoms, tear film parameters and ocular surface morphology was evaluated. METHODS: Patients with upper eyelid dermatochalasis were studied before and 3 months after upper eyelid blepharoplasty. Changes in ocular symptoms and physical examinations such as tear film break-up time, Schirmer test without local anaesthesia, impression cytology of the bulbar conjunctiva and inflammatory reaction were recorded. RESULTS: Subjective dry eye symptoms were found in 11 patients (46%) preoperatively and in five patients (21%) postoperatively. Objective dry eye signs were present in eight patients (33%) before blepharoplasty and in nine patients (38%) 3 months postoperatively. Morphological findings did not change within the observation period. The inflammatory reaction shown by impression cytology decreased postoperatively. CONCLUSION: Blepharochalasis may be associated with dry eye. Blepharoplasty might be a means of alleviating dry eye symptoms. The reason for this subjective alleviation might be due to reduced inflammatory reaction, changes in blink mechanism or the fact that patients are more confident about their appearance. PMID- 12780400 TI - Longterm results of one-site phacotrabeculectomy. AB - PURPOSE: To evaluate results of one-site phacotrabeculectomy and intraocular lens (IOL) implantation. METHODS: A retrospective study of a consecutive series of 62 eyes with cataract and glaucoma, which underwent primary one-site phacotrabeculectomy with IOL implantation. Mean follow-up time was 28 +/- 12.3 months (range 12-46 months). RESULTS: Preoperative mean intraocular pressure (IOP) (25.1 +/- 8.6 mmHg) dropped significantly, to 16.4 +/- 1.3 mmHg 1 year after surgery (p < 0.01). Mean logMAR best corrected visual acuity (VA) improved significantly from 0.74 +/- 0.49 to 0.22 +/- 0.28 (p < 0.01). The most common complications were fibrinous exudation (10%) and choroidal detachment (6%). In two eyes (3%) we successfully performed scleral flap revision to achieve IOP control. CONCLUSION: One-site combined surgery was a safe and effective procedure with good IOP control and visual rehabilitation. PMID- 12780401 TI - Effects of thick Tenon's capsule on primary trabeculectomy with mitomycin-C. AB - PURPOSE: To investigate the effects of thick Tenon's capsule on primary trabeculectomy with adjunctive mitomycin-C. METHODS: In this prospective interventional case series of 45 consecutive uncomplicated glaucoma patients, 45 eyes with thick Tenon's capsule underwent primary trabeculectomy with intraoperative mitomycin-C (0.4 mg/ml for 3 min). Success was defined as intraocular pressure (IOP) 13 s, a seizure energy index of >1000 u, and an electroencephalograph seizure length of > 24 s. "Adequate" overall ratings as determined by the rating scale were shown to correlate significantly with overall clinical improvement. The rating scale had extremely high interrater and test-retest reliability; and was relatively easy to use and time efficient. CONCLUSIONS: The use of the rating scale has the potential to help clinicians optimize ECT. While not perfect in any sense, it still gives clinicians good predictive qualities when used as a clinical guide. It is important that larger studies look at all the potential variables that may interact with seizure adequacy and clinical outcome. PMID- 12780472 TI - Bouffee delirante and ICD-10 acute and transient psychoses: a comparative study. AB - OBJECTIVE: ICD-10 acute and transient psychotic disorder (ATPD; F23) has one of its historical roots in the French concept of bouffee delirante. This study explores the empirical relationship of the two concepts. METHOD: During a 5-year period, all inpatients with ATPD were identified; the diagnosis of bouffee delirante was also determined. We systematically evaluated demographic and clinical features and carried out follow-up investigations at an average of 2.2 years after the index episode using standardized instruments. RESULTS: Forty-two (4.1%) of 1036 patients treated for psychotic disorders or major affective episode fulfilled the ICD-10 criteria of ATPD. Of these, only 28.6% also fulfilled the criteria of bouffee delirante. Patients with bouffee delirante were significantly younger than the remaining ATPD patients. Outcome parameters were generally more favourable for patients diagnosed with bouffee delirante than for ATPD patients without a concurrent diagnosis of bouffee delirante, but the difference was statistically significant only for occupational status. CONCLUSIONS: There are indications that a diagnosis of bouffee delirante carries a somewhat better prognosis than ATPD in general. However, the low frequency of operationally diagnosed bouffee delirante suggests that the criteria might be too narrow. PMID- 12780471 TI - A memory clinic at a geriatric hospital: a report on 577 patients assessed with the CAMDEX over 9 years. AB - OBJECTIVE: To report 9 years' experience of an Australian memory clinic using the -Cambridge Mental Disorders in the Elderly Examination (CAMDEX) assessment schedule, summarizing patient demographics, diagnoses at presentation and the utility of four instruments used in distinguishing patients with and without dementia. METHODS: All patients seen at the clinic between December 1989 and September 1998 were assessed using the CAMDEX. Diagnoses were determined according to criteria of the International Classification of Diseases, tenth edition (ICD-10). RESULTS: The mean age of 577 patients seen was 72.9 years and 60.8% were female. Over 40% fulfilled ICD-10 diagnostic criteria for dementia in Alzheimer's disease. A further 24% had another dementing illness. Only 28 patients were "normal". There was no significant difference in the ability of the 107-item Cambridge cognitive examination, the 30-item mini-mental state examination, the 10-item abbreviated mental test score and the 26-item informant questionnaire on cognitive decline in the elderly to differentiate dementia patients from those who were normal or had functional psychiatric disorders. The four cognitive screening tools had high correlations with one another (r = - 0.57 to 0.93). CONCLUSION: Patient demographics and diagnoses were similar to those found in other clinics. Most people who attended the memory clinic had significant cognitive or psychiatric disorders. PMID- 12780473 TI - Posttraumatic stress disorder: do electrical startle responses and thyroid function usefully supplement self-report? A study of Vietnam War veterans. AB - OBJECTIVE: To investigate the usefulness of electrical startle responses and thyroid function as supplements to self-report measures of posttraumatic stress disorder (PTSD). METHOD: Invitations were sent to all New Zealand Vietnam War veterans known to be living in North Canterbury; 50 responded and the 35 living in or near Christchurch were included. Self-report measures of PTSD (the Davidson Trauma Scale (DTS) and the Symptom Check List (SCL-90-R) ), an eye blink electrical startle response and thyroid function were measured. The DTS was re administered one to two weeks later to assess short-term test-retest reliability. Six months later the DTS and the electrical startle response were measured again. RESULTS: The veterans reported a wide range of PTSD severity, with 15/35 reporting prior diagnosis of PTSD. The DTS showed high short-term test-retest reliability (r = 0.93) and a moderate correlation after 6 months (r = 0.73). It also showed sensitivity to change; in one to two weeks the scores increased by nearly half a standard deviation, possibly because of an imminent "homecoming" march. The DTS and a PTSD scale from the SCL-90-R were highly correlated (r = 0.89). The total triiodothyronine (T3) to free thyroxine (T4) ratio measure of thyroid function correlated poorly with self-report (r < or = 0.27). The electrical startle response also correlated poorly with self-report (r < or = 0.26), showed low internal consistency between left and right sides (r = 0.43), and correlated 0.39 over six months. It was disliked by the veterans and had increased slightly at 6 month follow-up, perhaps because of sensitization. CONCLUSIONS: The DTS was reliable and correlated highly with the SCL-90-R PTSD scale. Neither thyroid function nor eye blink electrical startle correlated with each other or with self-report, and reliability was not good for electrical startle. These two measures do not appear to add anything useful to the assessment of PTSD. PMID- 12780474 TI - Dysexecutive syndrome and social adjustment in schizophrenia. AB - OBJECTIVE: This paper reports on a study designed to (i) assess levels of executive functions among patients suffering from schizophrenia; (ii) investigate associations between measures of executive functions and psychosocial adjustment; and (iii) examine the influence of psychopathology on the relationship between executive functions and psychosocial adjustment. Clear knowledge of executive functions and of their impact on social adjustment in patients with schizophrenia may play a decisive role in preparing and structuring appropriate outpatient care. METHOD: An extensive battery constituted of several tests developed for the assessment of executive functions was used in 38 inpatients with a DSM-IV diagnosis of schizophrenia. Psychosocial adjustment was assessed with a set of commonly used scales. RESULTS: Performance on measures of executive functions was heterogeneous among the patient population, with a subgroup performing within normal scores. A similar pattern was found on measures of psychosocial adjustment. Most of the executive measures were not dependent on demographic or clinical variables. A factorial analysis on measures of psychosocial adjustment yielded a one-factor model which showed inconsistent, and at most, weak to moderate correlations with executive functions. Regression analysis revealed that symptom levels accounted for two-thirds of the variance of psychosocial adjustment, and together with measures of executive functions for 91% of the variance. General and negative symptoms showed strong correlations with psychosocial adjustment. CONCLUSIONS: Our study supports the view that executive function is a heterogeneous construct with several subdimensions. Additionally, it suggests that symptom level has an important role as "rate limiting factor" on psychosocial adjustment and is partially mediated by executive dysfunction. Our results call for careful and detailed assessment in this patient population in order to establish appropriate treatment programmes such as cognitive remediation. PMID- 12780475 TI - What's the use of worrying? Its function and its dysfunction. AB - OBJECTIVE: To provide an overview of theory and research regarding the phenomenology and function of worry. METHOD: A systematic literature search focusing on conceptualizations of worry as a cognitive activity and worry research over the last 20 years was conducted. RESULTS: Worry can be both functional and dysfunctional. Disruption to daily functioning, perceived uncontrollability, pervasiveness and extensive coverage of worry themes (including "remote" events and the presence of meta-worry), all have specificity to more severe and dysfunctional worry. CONCLUSIONS: Research investigating both the relationship between worry and anxiety and the attributes of severe worriers, has brought attention to its role in fostering emotional disturbance and has clarified factors relevant to treatment. PMID- 12780476 TI - Awareness of illness in patients with bipolar I disorder with or without comorbid anxiety disorders. AB - BACKGROUND: The present study examined whether specific types of comorbid anxiety disorders, namely panic disorder (PD), social phobia (SP) and obsessive compulsive disorder (OCD) are differentially associated with course variables and insight into bipolar illness. METHOD: The sample consisted of 151 consecutively hospitalized patients with bipolar I disorder. They were assessed in the week prior to discharge using the Structured Clinical Interview for DSM-III-R (SCID P), the Brief Psychiatric Rating Scale (BPRS), the Global Assessment of Functioning Scale (GAF) and the Hopkins Symptom Checklist (HSCL-90). Level of insight was assessed with the Scale to assess Unawareness of Mental Disorders (SUMD). RESULTS: Of the 151 bipolar subjects, 92 had no PD, SP and OCD comorbidity, 35 had PD and 24 had SP and/or OCD. The three groups differed significantly on the current awareness of illness and treatment response scores and the retrospective awareness of illness and treatment response scores. Post hoc analyses revealed that, compared with bipolar patients without PD/SD/OCD and those with comorbid PD, patients with comorbid SP and/or OCD had better insight on current awareness of illness, current awareness of treatment response, retrospective awareness of illness and retrospective awareness of treatment response. The regression analysis showed that the presence of no panic type anxiety comorbidity was a predictor of good insight. CONCLUSIONS: These data indicate the value of identifying comorbid anxiety disorders in patients with bipolar illness. The results could be interpreted as evidence of discrete disorders within the bipolar spectrum, one that is characterized by, among other things, SP and/or OCD with good insight, another characterized by PD with poor insight. PMID- 12780477 TI - The excess cost of depression in South Australia: a population-based study. AB - OBJECTIVE: To establish excess costs associated with depression in South Australia, based on the prevalence of depression (from the Primary Care Evaluation of Mental Disorders (PRIME-MD)) and associated excess burden of depression (BoD) costs. METHOD: Using data from the 1988 South Australian (SA) Health Omnibus Survey, a properly weighted cross-sectional survey of SA adults, we calculated excess costs using two methods. First, we estimated the excess cost based on health service provision and loss of productivity. Second, we estimated it from loss of utility. RESULTS: We found symptoms of major depression in 7% of the SA population, and 11% for other depression. Those with major depression reported worse health status, took more time off work, reported more work performance limitations, made greater use of health services and reported poorer health-related quality-of-life. Using the service provision perspective excess BoD costs were AUD$1921 million per annum. Importantly, this excluded non-health service and other social costs (e.g. family breakdown, legal costs). With the utility approach, using the Assessment of Quality of Life (AQoL) instrument and a very modest life-value (AUD$50 000), the estimate was AUD$2800 million. This reflects a societal perspective of the value of illness, hence there is no particular reason the two different methods should agree as they provide different kinds of information. Both methods suggest estimating the excess BoD from the direct service provision perspective is too restrictive, and that indirect and societal costs ought be taken into account. CONCLUSIONS: Despite the high ranking of depression as a major health problem, it is often unrecognized and undertreated. The findings mandate action to explore ways of reducing the BoD borne by individuals, those affected by their illness, the health system and society generally. Given the limited information on the cost-effectiveness of different treatments, it would seem important that resources be allocated to evaluating alternative depression treatments. PMID- 12780478 TI - Validity and psychometric properties of the General Health Questionnaire-12 in young Australian adolescents. AB - BACKGROUND: The General Health Questionnaire (GHQ) is a measure of current mental wellbeing that has been extensively validated with adults. The instrument has also been used with adolescents. OBJECTIVE: (i) To assess the psychometric properties of the GHQ-12 among school students in grades 7-10; (ii) to validate it against other psychological tests; and (iii) to suggest a threshold score. METHOD: The survey was conducted in single sex and mixed schools from the state and private system in Perth, Western Australia. The survey contained the GHQ-12 and measures of anxiety, depression, self-esteem, stress, generalized self efficacy, social desirability and negative affectivity. RESULTS: There were 336 students (female 55%) with an age range of 11-15 years (median 13). The GHQ showed good internal consistency (alpha 0.88). Girls had higher mean GHQ-12 scores than boys (F (1,326) 15.0, p < 0.001) and scores for both genders increased with school grade (F (3,326) 4.2, p < 0.01). Multiple linear regression showed that depression, anxiety, self-esteem and stress were significant independent predictors of GHQ scores. The model accounted for 68% of the variance (adjusted R 2). Screening indices were calculated by comparison with a combined depression and/or anxiety category. Threshold scores of 13/14 for males and 18/19 for females appeared optimal. General Health Questionnaire scores were compared with two criterion groups: adolescents in hospital with alcohol or drug (AOD) related problems and those with problems not related to AOD use. Only the former group had significantly higher total scores. CONCLUSIONS: The GHQ-12 showed good structural characteristics and was appropriately correlated with other measures of related traits. Overall, the GHQ-12 appears to be a valid index of psychological wellbeing in this population and was considerably shorter than some of the other instruments. PMID- 12780479 TI - Relationship of meditation and psychosis: case studies. PMID- 12780480 TI - Publishing by Australasian psychiatrists. PMID- 12780481 TI - Psychosis associated with prescribed dexamphetamine use. PMID- 12780482 TI - Daily hassles, cortisol, and depression. PMID- 12780494 TI - Abnormal Pap test results and the rurality factor. AB - OBJECTIVE: To review the published socio-cultural determinants that put women in rural areas at greater risk of having an abnormal Pap test result. DESIGN: Literature review. CONCLUSION: An Australian woman is at increased risk of developing cervical dysplasia if she lives in a rural area, identifies as an Indigenous Australian or is from a non-English speaking country. Being aged over 50 years, increases the risk of cervical cancer but not cervical intra-epithelial neoplasia (CIN) 1, which peaks in the 20-24 year age group. Other connections with cervical dysplasia relate to a woman or her partner's sexual history, her nutritional status and possibly even her personality type. Smoking, social isolation, high negative life change events and having lived a 'troubled' life are all identified in the literature as increasing a woman's risk of having an abnormal Pap test result. Receiving an abnormal Pap test result often leads to feelings of fear, anxiety, guilt, shock, vulnerability, powerlessness, depression, shame and feeling dirty for the woman concerned. Compounding these negative feelings are social, practical and financial difficulties that have to be addressed. It is argued that social and medical models of health must complement each other to improve the well being of rural women experiencing an abnormal Pap result. PMID- 12780496 TI - Differences in access to health care services among adults in rural America by rural classification categories and age. AB - OBJECTIVE: To study differences in excess to health care services between different population groups in rural areas of the United States. DESIGN: Using data from the 1994 National Health Interview Survey and the 1991 Area Resource File, we examined the differences in excess with seven measures: having a regular source of care, having a usual place of care, having health insurance coverage, delaying medical care because of cost for all rural residents; number of doctor visits, number of hospital discharges and length of hospital stay per discharge for those who reported their health as being either poor or fair. Rural residents were classified by ages and grouped into four rural classification categories that were characterised along two dimensions: adjacent to a metropolitan statistical area (MSA) (yes/no) and inclusion of a city of at least 10,000 people (yes/no). SETTING: Rural areas. SUBJECTS: Rural populations. RESULTS: Residents aged 18-24 years had the worst access to services and the residents aged 65 years and over had the best access to services when measured by regular source of care, a usual place of care and health insurance status. Compared to those aged 50-64 years, residents aged 25-49 years were less likely to report having health insurance and more likely to report delaying seeking medical care because of costs. Rural residents who lived in a county adjacent to an MSA generally were less limited in access than those who lived in a county not adjacent to an MSA. CONCLUSIONS: Rural America is not a homogeneous entity in many aspects of the access to health care services. PMID- 12780495 TI - Straddling the pathway from paediatrician to mainstream health care: transition issues experienced in disability care. AB - OBJECTIVE: To identify the strengths and limitations of health care and related services provided to young adults with a disability during the period of transition from the care of a paediatrician to the mainstream health system. DESIGN: A descriptive design was used to address the study objectives. SETTING: Barwon and south-western region of Victoria. SUBJECTS: Twelve focus group discussions, with young adults with a disability, carers of young adults with a disability and health care service providers. Each focus group involved eight to 10 participants. RESULTS: The findings revealed a number of problems with the transition period. All participants acknowledged the supportive, coordinating role of the paediatrician. In the absence of this type of role, carers felt they lacked the knowledge and support to manage the adolescent with a disability. Communication problems between all service providers were identified as being problematical. The general lack of continuity of care between providers made it difficult for individuals to negotiate the transition period and increased the burden of care on carers. CONCLUSION: There is a need for policy makers to address these transition problems and develop appropriate services that improve the situation for young adults with a disability and their carers. PMID- 12780497 TI - Rural secondary school students living in a small community: their attitudes, beliefs and perceptions towards general practice. AB - OBJECTIVE: To study the attitudes and beliefs towards general practice of rural secondary school students, in particular, their perception of the barriers to the effective delivery of primary health care. SETTING: Students attending a government-funded secondary school in a rural community of less than 4000 people. SUBJECTS: 250 students in years 7-12 were surveyed, with 177 (71%) replying, 45% of these being male. DESIGN: Cross-sectional, self-administered questionnaire survey. RESULTS: The study found that most students value general practitioners (GPs) as valuable sources of health care, feel they have reasonable access to care and in most instances appear to attend appropriately for their health problems. Most felt their doctor would be an empathetic listener, would provide sufficient time to provide effective care and communicated in a language they could understand. Confidentiality issues were considered important, especially by females. CONCLUSION: Although students have very positive attitudes about general practice, the findings could be used to improve a number of GP services for young people in small rural communities. This is particularly so for presentations which require a high degree of confidentiality and skillful counselling, such as psychological problems and risk-taking behaviours. PMID- 12780498 TI - Self-reported patterns of health services utilisation: an urban-rural comparison in South Australia. AB - OBJECTIVE: To compare self-reported patterns of health service utilisation among residents of urban and rural South Australia. DESIGN, SETTING AND MAIN OUTCOME MEASURES: Secondary analysis of data generated by computer-assisted telephone interviews of 7377 adults done in 1995-6. Respondents were asked if they had used each of 18 different health services during the previous 12 months. Residence was classified in three ways: (1) capital city versus rest of the state, (2) by the Rural, Remote and Metropolitan Areas classification (RRMA) and (3) by the Accessibility and Remoteness Index for Australia classification (ARIA). RESULTS: General practitioner services were most frequently used, by approximately 89% of respondents. Only 4% reported not using any service. Comparing capital city with rest of the state, modest but statistically significant differences in utilisation (P < 0.01) were measured for nine services. In eight of these nine, utilisation was higher among rural residents. Analysing by RRMA, eight services were reportedly used differently and seven of these were the same as those identified from the capital city versus rest of state comparison. Across the five ARIA categories, six previously identified services were reported as being used differentially. Overall, rural residents had a higher than expected rate of moderate and high level of health service use. CONCLUSIONS: Self-reported use of a range of health services was broadly similar across urban and rural South Australia, with most cases of higher use were reported from rural areas rather than urban areas. Similar results were obtained when residence was classified in the three different ways. PMID- 12780499 TI - Rural origin medical students: how do they cope with the medical school environment? AB - Australia suffers from a well documented shortage of rural medical practitioners. In an attempt to increase recruitment, it has emerged that rural origin medical students are more than twice as likely as their urban colleagues to become rural practitioners. This has led to a wide range of programs aimed at increasing the number of rural students who apply for and gain entry into medical school. But how do rural students cope with the medical school environment? This paper was based on the results of a survey of senior medical students and looked at how rural students' fare with the medical school environment compared to their urban counterparts. It was concluded that government initiatives currently supporting rural medical students must be continued into the future and continuously evaluated to ensure that rural students have a positive learning experience in preparation for future rural practice. PMID- 12780500 TI - Wanted: trainees for rural practice. AB - To address the increasing need for rural health practitioners, Canada is trying various methods of medical education, including community based residency streams, additional skills training and teamwork models. This paper discusses some of the factors that may affect the effectiveness of these methods. PMID- 12780501 TI - Patterns of injury in indigenous Australians admitted to Cairns Base Hospital. AB - AIM: To study and compare the type and frequency of injury to indigenous and non indigenous patients admitted to the Orthopaedic Unit at Cairns Base Hospital. METHOD AND ANALYSIS: Relevant statistics for all patients were entered or calculated: demographics, length of stay, comorbidity and complications were available. After Variance Analysis, age-standardised rates of trauma were tested using confidence intervals. RESULTS: Of 2254 admissions for trauma in 32 months, 23% were indigenous (12% of the catchment population). Upper limb injuries were significantly higher for all indigenes, these males being admitted three times as frequently as non-indigenous males. Specific lesions showed greater differences and punch injuries were prominent in both indigenous sexes. Indigenous women were much more likely to be the victims of assault or fights than other groups. CONCLUSIONS: Personal injury costs are high in the indigenous community, in human and material terms. More resources should be directed towards injury prevention. PMID- 12780504 TI - Interventional radiology in the conservation of vascular access for hemodialysis. PMID- 12780505 TI - Study of the different types of actuators and mechanisms for upper limb prostheses. AB - Research in the area of actuators and mechanisms has shown steadily growing technological advances in externally activated upper limb prostheses. From among the actuators, advances include the use of piezoelectric materials, special metal alloys, polymers, and new motor applications, while the advances in mechanisms include mechanical designs based on the anatomy of the human hand and improvements in the way these components are combined. These efforts are aimed at meeting the need for anthropomorphic and functional prosthetic devices that enable patients to carry out basic daily tasks more easily and reduce the rejection rate of prostheses. This article technically discusses the several types of actuators and mechanisms, listing their main characteristics, applications, and advantages and disadvantages, and the current state of research in the area of rehabilitation of upper limb functions through the use of active prostheses. Comparisons of these devices are made with regard to the main criteria of construction and operation required to achieve optimal prosthetic performance. PMID- 12780506 TI - Shear stress related blood damage in laminar couette flow. AB - Artificial organs within the blood stream are generally associated with flow induced blood damage, particularly hemolysis of red blood cells. These damaging effects are known to be dependent on shear forces and exposure times. The determination of a correlation between these flow-dependent properties and actual hemolysis is the subject of this study. For this purpose, a Couette device has been developed. A fluid seal based on fluorocarbon is used to separate blood from secondary external damage effects. The shear rate within the gap is controlled by the rotational speed of the inner cylinder, and the exposure time by the amount of blood that is axially pumped through the device per given time. Blood damage is quantified by the index of hemolysis (IH), which is calculated from photometric plasma hemoglobin measurements. Experiments are conducted at exposure times from texp=25 - 1250 ms and shear rates ranging from tau=30 up to 450 Pa ensuring Taylor-vortex free flow characteristics. Blood damage is remarkably low over a broad range of shear rates and exposure times. However, a significant increase in blood damage can be observed for shear stresses of tau>or= 425 Pa and exposure times of texp>or= 620 ms. Maximum hemolysis within the investigated range is IH=3.5%. The results indicate generally lower blood damage than reported in earlier studies with comparable devices, and the measurements clearly indicate a rather abrupt (i.e., critical levels of shear stresses and exposure times) than gradual increase in hemolysis, at least for the investigated range of shear rates and exposure times. PMID- 12780507 TI - Extracorporeal gas exchange with the DeltaStream rotary blood pump in experimental lung injury. AB - In most severe cases of the acute respiratory distress syndrome, veno-venous extracorporeal membrane oxygenation (ECMO) can be used to facilitate gas exchange. However, the clinical use is limited due to the size and the concomitant risk of severe adverse events of conventionally-used centrifugal blood pumps with high extracorporeal blood volumes. The DeltaStream blood pump is a small-sized rotary blood pump that may reduce extracorporeal blood volume, foreign surfaces, contact activation of the coagulation system, and blood trauma. The aim of the present study was to test the safety and efficacy of the DeltaStream pump for ECMO in animals with normal lung function and experimental acute lung injury (ALI). Therefore, veno-venous ECMO was performed for 6 hours in mechanically ventilated pigs with normal lung function (n=6) and with ALI induced by repeated lung lavage (n=6) with a blood flow of 30% of the cardiac output. Gas flow with a FiO2 of 1.0 was set to equal blood flow. With a mean activated clotting time of 121 +/- 22 s, no circulatory impairment or thrombus formation was revealed during ECMO. Furthermore, free plasma Hb did not increase. In controls, hemodynamics and gas exchange remained unchanged. In animals with ALI, hemodynamics remained stable and gas transfer across the extracorporeal oxygenators was optimal, but only in 2 animals was a marked increase in PaO2 observed. CO2 removal was efficacious in all animals. We concluded that the DeltaStream blood pump may be used for veno-venous ECMO without major blood damage or hemodynamic impairment. PMID- 12780508 TI - Porcine stentless bioprostheses: prevention of aortic wall calcification by dye mediated photo-oxidation. AB - PURPOSE: Aortic wall calcification is problematic in stentless porcine valves. We evaluated the possible anticalcification effect of photo-oxidation on the aortic wall portion of porcine stentless bioprostheses. A comparison with glutaraldehyde fixed tissue was made. METHODS: Six Photofix and six Freestyle valves were implanted in juvenile sheep in pulmonary position. Valves were explanted after 3 or 6 months and examined macroscopically, by x-ray, light, and transmission electron microscopy. Calcium content was determined by atomic absorption spectrometry. RESULTS: The aortic wall portion of all Photofix valves remained free from calcification, while the wall portion of glutaraldehyde-fixed valves calcified strongly, both after 3 and 6 months. Calcium content of the aortic wall portion was: 0.71 +/- 1.27 in the Photofix valves versus 10.78 +/- 77.22 in the Freestyle valves (P < 0.05). CONCLUSION: Photo-oxidation of a porcine stentless valve prevents calcification not only in the cusps but also in the aortic wall portion. PMID- 12780509 TI - Prediction of single-pool Kt/v based on clinical and hemodialysis variables using multilinear regression, tree-based modeling, and artificial neural networks. AB - The impact of clinical and other variables on single-pool Kt/V (spKt/V) is unclear. The goal of this study was to identify clinical and hemodialysis treatment related predictors of spKt/V and use multilinear regression (LM), tree based modeling (TBM), and artificial neural networks (ANN) to predict actual spKt/V. When 602 hemodialysis records were analyzed, spKt/V correlated with urea reduction ratio (URR) (r=0.91) and weakly with other variables. When URR was excluded, both LM and TBM identified normalized protein equivalent of total nitrogen appearance (nPNA), prehemodialysis (HD) and post-HD weights, blood flow rate, and dialyzer surface area as predictors of spKt/V. LM identified sex, height, dialyzer ultrafiltration coefficient (Kuf), and duration of dialysis, while TBM identified the dialysis nurse code. Prediction algorithms were developed from a "training" dataset, and validated on a separate ("testing") dataset. Correlation coefficients of predicted spKt/V with measured spKt/V with and without nPNA respectively were 0.745 and 0.679 for LM, 0.6 and 0.512 for TBM, and 0.634 for ANN, which performed better without using nPNA. PMID- 12780511 TI - Successful MARS treatment in severe cholestatic patients with acute on chronic liver failure. AB - Extracorporeal liver devices have gained great attention as a complementary approach to liver transplantation in patients with acute on chronic liver failure. Among others, Molecular Adsorbent Recycling System (MARS) is a hemodiafiltration against albumin able to remove low molecular weight toxins. We aimed to validate the use of MARS in patients presenting with acute on chronic liver failure with severe cholestasis. We enrolled 7 patients with acute on chronic liver failure, presenting with bilirubin >25 mg/dl and hepatorenal syndrome and/or hepatic encephalopathy grade >II. Liver biochemistry, coagulation, blood cell count, electrolytes, ammonia, lactate, blood urea nitrogen, creatinine, bile acids, Fischer ratio, and encephalopathy grade were assessed before and after each MARS treatment. MARS can represent a safe therapeutic choice to achieve a quick improvement of neurological status, a hemodynamic stability, and a better clinical outcome. In particular, our encouraging results suggest that also, patients with severe cholestasis may represent in the future a good indication for MARS treatment. PMID- 12780510 TI - Modified DALI LDL-apheresis using trisodium citrate anticoagulation plus bicarbonate or lactate-buffered hemofiltration substitution fluids as primers. AB - BACKGROUND: DALI (direct adsorption of lipids) is the first LDL-apheresis technique able to adsorb low-density lipoprotein (LDL) and lipoproteina) directly from whole blood. In the standard procedure, acid citrate dextrose (ACD-A) is used as anticoagulation and the adsorber is rinsed with a specially manufactured priming solution (PS). Using neutral trisodium citrate (TSC) instead of ACD-A might improve the acid-base homeostasis during DALI apheresis; moreover, applying wholesale hemofiltration solutions instead of the special PS might avoid the use of two separate solutions for both priming before and reinfusion after the treatment, thus simplifiying the procedure. AIM: The present study was performed to test the effect of neutral (TSC) anticoagulation and of two different commercially available hemofiltration (HF) priming solutions on the efficacy and biocompatibility of DALI apheresis. MATERIALS AND METHODS: Five hypercholesterolemic chronic DALI patients were treated prospectively, on a weekly or biweekly basis, 3 times each by standard DALI-apheresis (A). by DALI using 4% TSC and bicarbonate-buffered HF BIC35-210 priming (B). as well as by DALI using 4% TSC and lactate-buffered HF 23 priming (C). After the sessions, the extracorporeal circuit (ECC) was rinsed with saline in study arm A and with the corresponding HF solutions in study arms B and C, respectively. RESULTS: Acute LDL-cholesterol reductions in the study arms A/B/C averaged 64/64/63%, for Lp(a) 62/64/62%, respectively (n=15). Clinically, all sessions were essentially uneventful and no clots were observed in the ECC. No major differences were found between the 3 study arms with respect to biocompatibility (elastase, C3a, thrombin-antithrombin, beta-thromboglobulin, bradykinin). CONCLUSION: DALI apheresis using TSC anticoagulation and HF solutions for both priming and reinfusion proved to be as safe and effective as the standard DALI apheresis. These modifications, however, further simplify the procedure. PMID- 12780512 TI - The effect of s-nitroso-glutathione on platelet and leukocyte function during experimental extracorporeal circulation. AB - Treatment with extracorporeal membrane oxygenation ECMO) is associated with side effects, e.g., blood cell consumption and activation. Our group has earlier shown that nitric oxide administered as a gas reduces platelet consumption and activation. In the present work we have studied the effect of the NO-donor S nitroso-glutathione GSNO) on platelets and leukocytes in an in vitro extracorporeal circuit. Two complete ECMO circuits were perfused with fresh heparinized human blood for 24 hours. GSNO was administered as a continuous infusion to one circuit at a rate of 0.7 mg/hour in four paired experiments and at a rate of 3.5 mg/hour in another four paired experiments. The other circuit was used as a control. Blood samples were withdrawn from both circuits before the start of the experiments and at 0.5, 1, 3, 12, and 24 hours of perfusion. The samples were analyzed for red blood cell count, leukocyte count, platelet count, platelet membrane expression of glycoproteins GP) Ib and GPIIb/IIIa, leukocyte membrane expression of cluster of differentiation CD) 11b/CD18, as well as plasma concentration of tumor necrosis factor TNF)-alpha, interleukin IL)-1beta, and IL 8. No difference in these parameters between the GSNO and the control circuit at any time point was assayed. In this study, no significant effect of GSNO on circulating platelets or leukocytes during experimental extracorporeal circulation could be shown. PMID- 12780513 TI - Heparin monitoring in sheep by activated partial thromboplastin time. AB - Heparin anticoagulation is utilized during and after vascular surgery in animals to reduce the risk of acute or chronic thromboembolic problems. In this study, we examined variation of activated partial thromboplastin time APTT) after the intravenous bolus IV bolus) and subcutaneous SC) heparin injection in order to monitor heparin therapy in sheep. Nine healthy sheep were assigned to 3 groups A, B, and C) according to their body weights: less than 40 kg, 40 to 80 kg, and more than 80 kg, respectively. All animals were treated with heparin 300 IU/kg body weight) through two routes, and the APTT, fibrinogen, and platelet count were measured before and every hour after treatment. This showed that the APTT was increased significantly between 1 to 4 hours after IV bolus injection and between 2 to 6 hours after SC injection P < 0.05). The APTT was returned to baseline values 6 and 10 hours after the respective treatments. The APTT in Group C was consistently higher than in Group A and B after heparin treatment by the two routes. The APTT ratio entered the subtherapeutic range 5 and 8 hours after IV bolus and SC injection, respectively. The APTT ratio was maintained in the therapeutic range for about 1 and 4 hours after IV bolus and SC injection, respectively. The highest APTT ratio in Group C after SC injection of heparin was significantly higher than that in Groups A and B P < 0.05). The mean platelet counts in Groups A, B, and C before the injection were 3197 +/- 365.6, 2886 +/- 78.2, and 1861 +/- 298.0 102/microL, respectively. The mean platelet count gradually decreased without significant variation after IV bolus and SC injection. These results produced elementary data for monitoring in sheep using APTT, and suggested that heparin should be administrated by the SC route at 4 hour intervals in order to remain in the therapeutic range, after an initial IV bolus dose. PMID- 12780514 TI - Honoring living legends. AB - From August 17 to 19, 2001, the University of Ottawa Heart Institute hosted the 6th Symposium of the World Artificial-Organ, Immunology and Transplantation Society (WAITS 2001). During WAITS 2001 a very special session entitled "Living Legends-Lessons Learned and Future Visions" was held. This session brought together giants in the field who had pioneered organ transplantation, cardiovascular surgery, pacemakers, and artificial hearts, among many other major contributions. The purpose of the session was to both honor and learn from those who have contributed greatly to the major scientific and technological advances in our field during the twentieth century. PMID- 12780515 TI - In vitro and in vivo study of ion-implanted collagen for the substrate of small diameter artificial grafts. AB - Ion implantation into the collagen-coated inner surface of the grafts was performed and evaluated in vitro and in vivo to develop small diameter artificial vascular grafts. He+ ion implanted collagen-coated grafts with a fluence of 1 x 1014 ions/cm2 inhibited platelet adhesion and demonstrated patency for 240 days in an animal study. The platelet adhesion test using platelet rich plasma (PRP) showed antithrombogenicity at the fluence of 1 x 1014 ions/cm2. Washed platelet adhesion test showed thrombus formation at the fluence of 1 x 1014 ions/cm2. The results suggested that plasma protein adsorption onto the ion-implanted collagen significantly improved performance of these synthetic grafts. PMID- 12780516 TI - Trileaflet valve for VAD use with purged sinus. AB - Clinical applications of ventricle assist devices continue to be problematic due to thromboembolic complications. The problem originates mainly at the valves, which are usually made of an antithrombogenic material, such as cross-linked bovine pericardium. However, wherever the blood flow is stagnant or forms a recirculation region, a thrombus is likely to form. A similar blood flow is found in the space between the housing of the valve and the leaflets, the so-called valve sinus. Consequently, thrombi are often generated in this region. The novel valve design presented in this article avoids the formation of stagnant flow in the valve sinus during systole by a purge flow. This flow is taken from the main flow through the valve and is directed into each sinus region. The effect is achieved by perforation of the valve sinus with a small orifice at the bottom of the sinus. The purge flow effect is investigated with the computational fluid dynamics (CFD) method. The simulation shows that the purge flow effectively increases flow in the valve sinuses. PMID- 12780517 TI - Activated leukocytes adsorbed on the surface of an extracorporeal circuit. AB - Biocompatibility of extracorporeal circuits has mostly been investigated by sampling blood in circulation. However, a proportion of activated leukocytes leave circulation by sticking to the circuits, and might affect the circuit biocompatibility. To reveal these characteristics, we eluted the adsorbed leukocytes from circuits used for 6 patients by washing with ethylenediaminetetraacetic acid (EDTA)-HEPES buffer. 1.3 x 109 leukocytes, representing 5 to 10% of the circulating leukocytes, were collected. Most were neutrophils expressing CD11b antigens as high as those in circulation during cardiopulmonary bypass. Adsorbed monocytes expressed tissue factor higher than those in circulation (P < 0.05). An elution procedure by EDTA itself induced only a minimal activation of the leukocytes. These results indicate that extracorporeal circuits adsorb a large number of activated neutrophils and a small number of highly thrombogenic monocytes during their use. Although our data is limited, this elution procedure appears useful to investigate the biocompatibility of extracorporeal circuits. PMID- 12780518 TI - Prognostic factors in laryngeal carcinoma: the role of apoptosis, p53, proliferation (Ki-67) and angiogenesis. AB - Even though the roles of different known or suggested prognostic factors in laryngeal cancer have been studied in detail, clinical stage at time of diagnosis and anatomic subsite of the tumour remain the only practical predictors of clinical outcome and offer the only guidelines in the planning of treatment. In this study, the relative roles of known demographic and clinical prognostic factors, in addition to four histopathological factors, were evaluated in a sample of 100 laryngeal carcinoma patients with multivariate analysis using the Cox regression model. In addition to advanced stage (stage III-IV) (relative hazard of death (HR) 8.9, p=0.01) and supraglottic disease (HR 5.6, p=0.02), high apoptotic index (HR 11.1, p=0.05) was significantly associated with poor survival. Cell proliferation, p53 and angiogenesis did not significantly affect the prognosis. In the future, high degree of apoptosis could be used to identify patients with poor prognosis in laryngeal cancer. PMID- 12780519 TI - VEGF in 105 pheochromocytomas: enhanced expression correlates with malignant outcome. AB - Pheochromocytomas are rare sympathoadrenal tumors that are highly vascular. Their malignancy is extremely difficult to estimate on the basis of histopathological features. Vascular endothelial growth factor (VEGF) is one of the most important angiogenic factors involved in both tumor growth and metastasis. In our search for new prognostic markers, we investigated the expression of VEGF in normal adrenal gland, in 105 primary pheochromocytomas, and in 6 metastases by using immunohistochemistry and Northern blot analysis. We also calculated the microvessel density of these tumors by staining the endothelial cells with monoclonal CD34 antibody. VEGF messenger ribonucleic acid was found in all pheochromocytomas studied. Immunohistochemically, VEGF was not found in normal adrenal medullary cells. Interestingly, all malignant pheochromocytomas (n=8), regardless of their primary location, had strong or moderate VEGF immunoreactivity, while most benign adrenal pheochromocytomas (26 of 37, 70.3%) were either negative or only weakly positive. The staining was heterogenous in extraadrenal pheochromocytomas as well as in a group of tumors that had histologically suspicious features but had not metastasized, here called borderline tumors (n=29). The microvessel density varied greatly in all of the tumor groups, and no statistical difference was found between these groups. Here we report moderate to strong VEGF expression in malignant pheochromocytomas, and negative or weak expression in benign adrenal pheochromocytomas. Normal medullary cells are immunohistochemically negative. Thus, low VEGF expression in pheochromocytomas favors a benign diagnosis. PMID- 12780520 TI - Genetic alterations of the HCCS1 gene in Korean hepatocellular carcinoma. AB - We analyzed the gene mutations and loss of heterozygosity (LOH) of the HCCS1 gene using intragenic polymorphic markers in a series of 88 primary HCCs. We found two sequence variations at exon 5 and 14 in both normal and tumor DNAs of case 50 and 51, respectively. The variation in case 50 led to a reading frameshift and a premature stop (TGA) at codon 125 and case 51 showed amino acid change at codon 448 (Val-->Ala, GTG-->GCG). Interestingly, these variations were not found in peripheral lymphocytes of 69 normal individuals and 227 cancer patients (86 HCC, 75 unselected gastric cancer, and 66 breast cancer), suggesting that these two variations are mutation, not polymorphism. In addition, we found 14 novel intragenic polymorphic sites in the HCCS1 gene. Thirty-two (47%) of sixty-eight informative cases showed allelic loss at at least one or more intragenic polymorphic sites, but there was no significant relationship between the frequency of LOH and clinicopathologic parameters. These results suggest that mutation of the HCCS1 gene might not be a main inactivation mechanism in the development of Korean HCC and that the HCCS1 gene might be involved in acceleration of the tumorigenic process in Korean HCC. PMID- 12780521 TI - Comamonas testosteroni meningitis in a patient with recurrent cholesteatoma. AB - Comamonas testosteroni, a lesser-known member of the genus, has shown little apparent capacity for causing infections in humans. We here present a case of purulent meningitis due to C. testosteroni, which occurred in a patient who had recurrent cholesteatoma. Ceftriaxone treatment was not effective in this patient even though in vitro the bacteria were susceptible to the drug. The patient responded well to meropenem therapy. PMID- 12780522 TI - Effects of Ocimum gratissimum L essential oil at subinhibitory concentrations on virulent and multidrug-resistant Shigella strains from Lagos, Nigeria. AB - Ocimum gratissimum leaf extracts have been extensively demonstrated to be effective against the various aetiologic agents of diarrhoea, including Shigellae. However, the mechanism of the shigellocidal action of this plant remains to be understood. This study investigated the effects of O. gratissimum essential oil (EO) at subinhibitory concentrations of 0.75 and 1.0 microg/ml on virulence and multidrug-resistant strains of 22 Shigella isolates from Nigeria. Compared with untreated Shigella strains, O. gratissimum EO caused significant decreases (p<0.01) in extracellular protease activity, o-lipopolysaccharide rhamnose content and incidence of invasiveness mediated as keratoconjunctivitis in guinea pig. The disparity in extracellular protease activity and o lipopolysacharide rhamnose between the two treatment groups was also found to be significant (p<0.05), suggesting greater anti-virulent effects of O. gratissimum oil at 1.0 microg/ml. Antibiotic susceptibility testing revealed that the EO of O. gratissimum reduced the MICs of antibiotics to which Shigellae showed resistance by 9.8-53.1% and fluoroquinolones by 18.2-45.5%. The results of this study strongly suggest inhibition of extracellular protease and expression of O LPS rhamnose in Shigellae by O. gratissimum EO. The future use of O. gratissimum- antibiotic combinations as a therapeutic measure against shigellosis is discussed. PMID- 12780523 TI - Tumour-type-specific capillary endothelial cell stainability in malignant B-cell lymphomas using antibodies against CD31, CD34 and Factor VIII. AB - The microvessel density (MVD) was assessed in lymph nodes infiltrated by diffuse large B-cell lymphomas, mantle cell lymphomas, chronic lymphatic leukemia and follicular lymphomas, and in lymphadenitis. Serial sections of formalin-fixed and paraffin-embedded tissue were stained with antibodies against CD31, CD34 or Factor VIII. Using light microscopy and computerised image analysis, the number and size of individual immunostained vessel profiles within a preselected area size range corresponding to capillaries, postcapillary venules, small collecting venules and small arterioles were determined. A significantly larger number of vessels were registered following staining with anti-CD34 than with anti-CD31 or anti-Factor VIII. Moreover, among the smallest capillary-sized vessel profiles in all lesion types, there was a selective relative loss of stainability of anti CD31 and anti-Factor VIII, resulting in a substantial total loss of visualised capillary-sized vessels compared with anti-CD34. In fact, the number of non detected capillaries following staining with anti-CD31 and anti-Factor VIII was significantly tumour type specific. These findings influence how we evaluate MVD data in B-cell lymphomas and possibly also other tumour types, as well as data relating to capillary endothelium-related functional variables of proliferation, apoptosis and maturation when different double-labelling immunohistochemical techniques are used and different tumour types are analysed. PMID- 12780524 TI - Mutational analysis of Fas ligand gene in human non-Hodgkin lymphoma. AB - Among the systems triggering apoptosis, the Fas-Fas ligand (FasL) system is recognized as a major pathway for the induction of apoptosis in cells and tissues. Ligation of Fas by either an agonistic antibody or FasL transmits a 'death signal' to the target cell, potentially triggering apoptosis. Alterations of genes along the Fas-mediated apoptosis pathway have been reported in many human cancers. However, there have been no data regarding FasL gene mutations in human cancers. We hypothesized that FasL gene mutation might be involved in the development of non-Hodgkin lymphoma (NHL). In this study, we analyzed the entire coding region of the FasL gene for the detection of somatic mutations in a series of 111 NHLs and found that one tumor had a FasL gene mutation in the cytoplasmic domain. To evaluate the functional alterations of the mutant in apoptosis, we overexpressed the mutant in 293T cells, but couldn't find any significant loss of cell death compared to the wild-type FasL. Together, these data suggest that FasL is occasionally mutated in human NHL and that FasL mutations appear to play no role in the pathogenesis of the vast majority of NHLs. PMID- 12780525 TI - Heat shock preconditioning modulates proliferation and apoptosis after superficial injury in isolated guinea pig gastric mucosa via an eicosanoid and protein synthesis-dependent mechanism. AB - AIM: In restitution after superficial injury of the gastric mucosa, the epithelial continuity is restored by cellular migration. We have shown that heat shock preconditioning inhibits restitution after superficial injury. This study investigates the effect of heat shock preconditioning on tissue proliferation and apoptosis. EXPERIMENTAL DESIGN: Paired guinea pig gastric mucosae were mounted and perfused in Ussing chambers (37 degrees C). After heat shock preconditioning (42 degrees C) (30 min) and normothermic recovery (37 degrees C) (150 min) or normothermic perfusion, a superficial injury was induced by luminal exposure to 1.25 mol/L NaCl (5 min) followed by a 3 h restitution. During perfusion, the mucosa was exposed to 30 micromol/L arachidonic acid (AA) to enhance heat shock response, to 50 micromol/L quercetin (Q) to inhibit the metabolism of arachidonic acid via lipoxygenases, to 50 micromol/L indomethacin (In) to inhibit the metabolism of arachidonic acid via cyclo-oxygenases, or to 150 micromol/L cycloheximide (CHX) to inhibit de novo protein synthesis. After the experiment the mucosa was prepared for immunohistochemical analysis of the expression of Mib 1 proliferation antigen and pro-apoptotic protein Bax. RESULTS: Heat shock decreased Mib-1/Bax ratio and this effect was maintained after superficial injury and exposure to Q, to AA+CHX or to In+CHX. Exposure to CHX, to AA, to In+Q, to In+AA, In+AA+Q or to In+AA+CHX, however, blocked the effect of heat shock preconditioning. The decreasing effect of heat shock preconditioning on Mib-1/Bax ratio could be reversed by exposure to AA+Q or to In. CONCLUSION: The heat preconditioning-induced effects on the mucosa are reversible and sensitive to exogenous pharmacological modulation. Heat shock preconditioning inhibits proliferation of superficially injured isolated gastric mucosa by a mechanism involving eicosanoid pathways and de novo protein synthesis. PMID- 12780526 TI - A study on forensic samples of Bartonella spp antibodies in Swedish intravenous heroin addicts. AB - Infection with Bartonella, an emerging bacterial pathogen which often affects immunodeficient patients, has been reported in Sweden over the past few years, with a high seroprevalence of B. elizabethae. A high prevalence of antibodies against B. elizabethae has also been found in urban intravenous drug users in the USA. Using immunofluorescence, we retrospectively examined serum samples taken at autopsy from 59 Swedish intravenous drug addicts from the Stockholm area for evidence of antibodies against 6 pathogenic strains of Bartonella. The 59 addicts died following heroin injection during the years 1987-1992 and include 24 individuals (41%) who were additionally HIV-positive. An overall seropositivity rate for Bartonella spp. of 39% (23/59) was found with the following antigenic reactivities: B. elizabethae, 39% (23/59); B. grahamii, 3% (2/59); B. henselae (Houston-1), 14% (8/59); and B. quintana, 3% (2/59). There were no positive reactions for B. henselae (Marseille) or B. vinsonii subsp. vinsonii. The Bartonella-seropositive cases included 11/23 (48%) individuals who were HIV positive. Subacute to chronic myocarditis was seen in 2/11 microscopically investigated Bartonella-seropositive cases that were HIV-negative and in 1/14 seronegative cases. No cases of endocarditis or other common manifestations of Bartonella infection were found. An overall Bartonella seropositivity of 21% (9/44) was observed in control forensic autopsy samples. PMID- 12780527 TI - Two-year follow-up of Helicobacter pylori infection in C57BL/6 and Balb/cA mice. AB - Helicobacter pylori infection is associated with chronic gastritis, peptic ulcer disease, gastric adenocarcinoma and MALT lymphoma. We previously found high-grade lymphoma after 13 months' H. pylori infection in C57BL/6 mice. In this study we followed H. pylori infection by three different isolates in C57BL/6 and Balb/cA mice for 23 months. Six-week-old C57BL/6 and Balb/cA mice were infected with H. pylori strains 119p (CagA+, VacA+), SS1 (CagA+, VacA+) and G50 (CagA-, VacA-). Mice were followed at 2 weeks, 10 weeks and 23 months post-inoculation (p.i.) by culture, histopathology and serology. Strain G50 was only reisolated from mice 2 weeks p.i. There was no difference in colonization between strain 119p and SS1 at 10 weeks p.i., whereas SS1 gave 100% colonization versus 119p gave 50% 23 months p.i. Interestingly, the inflammation score was higher in mice infected with strain 119p than with SS1 10-week p.i., and there were lymphoepithelial lesions in mice infected with strain 119p and G50 but not with SS1 at 23 months post infection. Eight mice infected with strains 119p and G50 developed gastric lymphoma (grade 5 and 4). One C57BL/6 mouse infected with strain 119p developed hepatocellular carcinoma after 23 months. Immunoblot showed specific bands of 26 33 kDa against H. pylori in infected mice, and two mice infected with strain SSI reacted with antibodies to the 120 kDa CagA toxin. CONCLUSION: A reproducible animal model for H. pylori-induced lymphoma and possibly hepatocellular carcinoma is described. Strain diversity may lead to different outcomes of H. pylori infection. PMID- 12780528 TI - Expression of heat-shock proteins hsp27, hsp70 and hsp90 in malignant epithelial tumour of the ovaries. AB - Recently, considerable attention has been focused on the role of the small heat shock protein group hsp27, hsp70 and hsp90 in the clinical outcome of several malignancies. However, conflicting data exist regarding the prognostic role of hsp27 expression in ovarian carcinoma, and the prognostic significance of hsp70 and hsp90 expression still remains unknown in these tumours. The purpose of this study was to investigate immunohistochemically whether hsp27, hsp70 and hsp90 expression was associated with clinicopathological parameters and survival in 52 epithelial ovarian carcinomas. Chi-square test, Kaplan-Meier and Cox regression analysis were used for statistical analysis. Among clinicopathological parameters, hsp27, hsp70 and hsp90 expression was only correlated with FIGO stage; hsp70 and hsp90 positivity failed to detect survival. However, the overall survival rate of patients with hsp27 expression was 13%, which was significantly worse than that of patients without hsp27 expression (47%) (p<0.01). The prognosis was also adversely affected by FIGO stage (p<0.01) and presence of ascites (p<0.01). In multivariate analysis, hsp27 expression and FIGO stage were independent prognostic variables. Our results indicate that hsp70 and hsp90 expression had no prognostic relevance in epithelial ovarian carcinomas. However, hsp27 expression and FIGO stage in these tumours could be reliable indicators of prognosis. PMID- 12780529 TI - Experimentally induced hypo- and hyper-thyroidism influence on the antioxidant defence system in adult rat testis. AB - The objective of the present experiment was to study the effect of thyroid hormone on the antioxidant defence system of rat testis. Hypothyroidism induced in rats by 6-n propyl 2-thiouracil (PTU) treatment resulted in a reduction in body weight, seminal vesicle and ventral prostate gland. A further decrease in the weight of seminal vesicle was recorded following administration of T3 to hypothyroid rats. The oxidative stress parameters such as hydrogen peroxide and protein carbonyl content increased in the crude homogenate of testis of hypothyroid rats. T3 administration to hypothyroid rats resulted in no further change in the hydrogen peroxide level but the protein carbonyl content further elevated in the crude homogenate of testis. No significant change was observed in the endogenous lipid peroxidation level of the crude homogenate of testis whereas the FeSO4/ascorbic acid induced lipid peroxidation level decreased in hypothyroid rats and did not change further by T3 administration. Although the reduced glutathione level in the crude homogenate of testis did not change following hypothyroidism, oxidized glutathione level increased. The reduced and oxidized glutathione level decreased and increased, respectively following T3 administration to hypothyroid rats in comparison with PTU-treated rats. Activities of superoxide dismutase and catalase decreased in the post mitochondrial fraction (PMF) of testis of hypothyroid rats. T3 injection to PTU treated rats resulted in an elevation in the level of catalase activity only. The activity of glutathione peroxidase in the PMF of testis elevated in the hypothyroid rats and reduced following T3 treatment to hypothyroid rats. The results of the present study suggest that any alteration in the thyroid hormone level in the body affects the antioxidant defence system of testis of adult rats and, thereby, may affect the physiology of testis through oxidative stress. PMID- 12780531 TI - Megalospermatocytes in the human testis exhibit asynapsis of chromosomes. AB - Testis biopsies of three infertile patients were identified, which showed a predomination of megalospermatocytes in the seminiferous tubules. Megalospermatocytes are very large primary spermatocytes indicating a spermatogenic arrest. Because of the high percentage of these germ cells it was possible to apply a whole-mount spreading technique to investigate the chromosomal pairing behaviour in prophase I of meiosis. It could be shown that most of the megalospermatocytes exhibited extensive chromosomal asynapsis, suggesting that a characteristic meiotic disorder may give rise to reduced fertility, or even infertility. PMID- 12780530 TI - Immunohistochemical distribution of S-100 protein and subunits (S100-alpha and S100-beta) in the swamp-type water buffalo (Bubalus bubalis) testis. AB - The distribution and localization of S-100 protein (S-100) and its subunits (S100 alpha and S100-beta) in the testis of swamp-type water buffalo were investigated using immunohistochemistry. S-100 was detected in the Sertoli cells in the convoluted seminiferous tubules, modified Sertoli cells lining the terminal segment of the seminiferous tubules and in the intratesticular excurrent ducts (straight tubules and rete testis). S100-beta showed the same distribution and localization with that of S-100. However, the cytoplasmic extension of the Sertoli cells in S100-beta staining showed less staining intensity compared with that of S-100. S100-alpha showed a positive staining only in the modified Sertoli cells of the terminal segment of the seminiferous tubule. Endothelial cells of blood vessels were also positive with the proteins while the Leydig and spermatogenic cells showed a negative reaction. The localization of S-100 in the testis of the water buffalo was in parallel with that of other artiodactyls which supports the hypothesis that this protein is a multifunctional protein. S100-beta in the Sertoli cells suggests that this protein is involved in establishing blood testis barrier. Its presence in the modified Sertoli cells and in the epithelium of the excurrent ducts suggest secretory and absorptive function, respectively. Meanwhile, S100-alpha, which was detected only in the modified Sertoli cells, is involved in the secretory activity of these cells that are related to exocrine function. PMID- 12780532 TI - Significance of inflammation on standard semen analysis in chronic prostatitis/chronic pelvic pain syndrome. AB - The impact of defined urogenital inflammations on standard ejaculate parameters is still a matter of controversial debate. Basic spermiogram parameters has been analysed in patients with inflammatory and noninflammatory chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS-NIH IIIA/IIIB) with regard to indicators of inflammation in prostatic secretions and/or the ejaculate. A total of 112 consecutive patients symptomatic for chronic pelvic pain were included in the study. All of them underwent a 'four glass-test' including leukocyte determination in expressed prostatic secretions followed by ejaculate analysis according to WHO. The analysis included pH, volume, total sperm count, sperm density, motility, morphology (Shorr stain), vitality (eosin stain), and counting of peroxidase positive leukocytes (PPL). Patients were first subgrouped according to elevated leukocyte counts in prostatic secretions, and then according to the number of PPL in semen. Leukocytes neither in the prostatic secretions nor in the ejaculate were associated with reduced standard semen parameters. Our data supports previous results that elevated leukocyte counts in prostatic secretions and in ejaculate, as indicators of inflammation have no negative impact on total sperm count, sperm density, motility, morphology, and sperm vitality in patients with CP/CPPS. PMID- 12780533 TI - Virus infections in disorders of the male reproductive system. PMID- 12780534 TI - Classification and diagnosis of prostatitis: a gold standard? AB - The National Institutes of Health Classification System for prostatitis has now been accepted by the North American and International urology community. This categorization system consists of category I (acute bacterial prostatitis), category II (chronic bacterial prostatitis), category III (chronic prostatitis/chronic pelvic pain syndrome - CP/CPPS) and category IV asymptomatic inflammatory prostatitis. The evaluation of a patient with category I and category II bacterial prostatitis consists of history and physical examination and urine culture for lower urinary tract localization cultures, respectively. The clinical tests for the evaluation of CP/CPPS can be classified as mandatory, recommended and optional. Mandatory evaluations include history and physical examination, urinalysis and urine culture. Recommended evaluations include lower urinary tract localization tests, symptom index, flow rate, residual urine determination and urine cytology. Optional evaluations include semen analysis and culture, urethral swab, urodynamics, cystoscopy, imaging, and prostate specific antigen determination. The physician must individualize a rational diagnostic strategy for each patient. There is no 'gold standard' for the diagnosis and evaluation of patients presenting with prostatitis. PMID- 12780535 TI - Assisted reproductive techniques: risks, contraindications, prognostic factors, therapeutic strategies. PMID- 12780536 TI - Limits of infertility surgery - gynaecological view. PMID- 12780537 TI - Pre-implantation genetic diagnosis - possibilities and pitfalls. PMID- 12780538 TI - Assisted reproduction: genetic aspects - risk of malformations - pre-treatment counselling. PMID- 12780539 TI - From the oocyte to the blastocyst. PMID- 12780540 TI - Is there an increased genetic risk of old fathers towards their offspring? PMID- 12780541 TI - Current aspects of donor insemination. PMID- 12780542 TI - Limits of microsurgical refertilization under urological aspects. PMID- 12780543 TI - Therapeutic strategies in severe male factor infertility. PMID- 12780544 TI - Cryopreservation of ovarian tissue to preserve female fertility - state of the art. PMID- 12780545 TI - Limitations for ICSI, MESA, TESE? - experiences from the IVF centre in Giessen. PMID- 12780546 TI - Perspectives in the diagnosis of testicular biopsies using molecular biological techniques. PMID- 12780547 TI - Reproductive medicine after menopause. PMID- 12780548 TI - Gynaecological malignancies and infertility. PMID- 12780549 TI - Epididymal sperm retrieval: indications, risks, and outcome. PMID- 12780550 TI - HIV-infected people who wish to have children - chances and limits of assisted reproductive techniques. PMID- 12780552 TI - The clinical importance of alloantibody-mediated rejection. PMID- 12780553 TI - Role of nitric oxide synthase (NOS) during ex vivo warm kidney perfusion prior to renal transplantation. PMID- 12780554 TI - Less is more: maintenance minimization as a step toward tolerance. PMID- 12780555 TI - Complement C4d in graft capillaries -- the missing link in the recognition of humoral alloreactivity. AB - Staining of C4d in graft capillaries has emerged as a useful method to detect antibody-mediated rejections in situ. Demonstration of capillary C4d has provided substantial clinical results and allows several conclusions: Antidonor antibodies (preformed or produced de novo) activate complement directly in the graft. Capillary C4d is present in about 30% of biopsies with acute and chronic rejections and separates rejections with a humoral component from 'pure' cell mediated rejections. Recognition of humoral alloreactivity is important, since effective treatment is now available. Since capillary C4d can appear and disappear at any time post transplantation, every transplant biopsy should be tested. Capillary C4d is now incorporated in the 'Banff classification'. The incidence of C4d-positive cases will probably decline because of the 'routine' application of potent immunosuppressants, including mycophenolate mofetil, that can inhibit antibody production. Presensitization, however, will remain a potential threat to allografts. PMID- 12780556 TI - Utility of intravenous immune globulin in kidney transplantation: efficacy, safety, and cost implications. AB - Intravenous immunoglobulin preparations (IVIG) are known to be effective in the treatment of various autoimmune and inflammatory disorders into their immunomodulatory, immunoregulatory, and anti-inflammatory properties. Recently, IVIG has been utilized in the management of highly sensitized patients awaiting renal transplantation. The mechanisms of suppression of panel reactive antibodies (PRA) in patients awaiting transplantation are currently under investigation and appear to be related to anti-idiotypic antibodies present in IVIG preparations. In this review, the various immunomodulatory mechanisms attributable to IVIG and their efficacy in reducing PRAs will be described. In addition, the use of IVIG in solid organ transplant recipients will be reviewed. The adverse events, safety considerations, and economic impact of IVIG protocols for patients awaiting solid organ transplantation will be discussed. PMID- 12780557 TI - Humoral theory of transplantation. AB - According to the humoral theory of transplantation, antibodies cause allograft rejection. Publications are cited showing that antibodies: (1). cause hyperacute kidney rejection, (2). lead to C4d deposits associated with early kidney graft failures, (3). are a good indicator of presensitization leading to early acute rejections, (4). were present in 96% of 826 patients who rejected a kidney graft, (5). are associated with chronic rejection in 33 studies of kidney, heart, lung and liver grafts, and (6). in three studies, appeared in the circulation BEFORE evidence of bronchiolitis obliterans in lung transplants, and BEFORE kidney rejection. In addition, a prospective cooperative study of 1629 patients in 24 centers demonstrated that antibodies foretold subsequent failures after a follow up period of 6 months (p = 0.05). The specificity of antibodies detected in the serum of rejecting patients were often not donor specific, presumably because they were absorbed by the rejecting organ. If the humoral theory is accepted, even provisionally, transplanted patients who have antibodies could be treated with immunosuppression until the antibodies disappear to determine whether chronic rejection can be blocked. If successful, in patients who do not have antibodies, immunosuppression could be reduced until antibodies appear. PMID- 12780558 TI - NOS: the underlying mechanism preserving vascular integrity and during ex vivo warm kidney perfusion. AB - Research involving metabolically active and functioning organs, maintained ex vivo in culture-like conditions, could provide numerous opportunities for medical innovations and research. We report successful perfusion of isolated canine and human kidneys ex vivo at near physiologic temperature for 48 h. During the perfusions parameters of metabolism and function remained stable. Nitric oxide synthase (NOS) was identified as the underlying mechanism preserving vascular integrity. Most importantly, when the canine kidneys were reimplanted there was immediate normal renal function. This report highlights the potential significance of whole organ culture using a warm temperature ex vivo perfusion and discusses medical applications that could be developed. PMID- 12780559 TI - CD154 regulates primate humoral immunity to influenza. AB - Current methods of immunosuppression for the purposes of allowing solid organ transplantation in humans are broadly inhibitory and thus are associated with an increased risk of opportunistic infections and neoplasia. We have shown previously that a selective blockade of CD40-CD154 interactions during heart transplantation in cynomolgus macaques significantly delays immune-mediated graft injury. Here, we determined the effect of anti-CD154 mAb therapy on primate serologic responses to immunization with influenza virus hemagglutinin (HA), a T cell-dependent Ag. We found that CD154 blockade attenuated primary and secondary serum Ab responses of IgM and IgG isotypes to influenza, even when anti-CD154 treatment was discontinued prior to reimmunization. These findings show that in primates CD40-CD154 interactions are necessary for both primary and secondary Ab responses to viral Ags. Furthermore, the data suggest that viral Ag stimulation of primates in the absence of CD154 stimulation may have a tolerizing effect on that Ag. PMID- 12780560 TI - Dendritic cell subset ratio in peripheral blood correlates with successful withdrawal of immunosuppression in liver transplant patients. AB - Human dendritic cell (DC) subsets appear to play distinct roles in the induction and regulation of immune responses. While monocytoid DC (DC1) induce T-helper (Th) 1-type responses, plasmacytoid DC (DC2) have been reported to selectively induce Th2 responses. In blood, their precursors (p) can be identified as HLA-DR+ lineage- cells that are further characterized as CD11c+ CD123-/lo (IL-3Ralpha /lo) (pDC1) or as CD11c- CD123hi (pDC2) by rare event, flow cytometric analysis. We compared the incidences of pDC1 and pDC2 in peripheral blood mononuclear cell populations isolated from normal healthy controls and from 3 groups of clinically stable liver transplant patients. Group A had been successfully withdrawn from immunosuppression, whereas group B were undergoing prospective drug weaning and on minimal anti-rejection therapy. In group C, drug withdrawal had either failed or never been attempted and patients were on maintenance immunosuppression. Assessment of DC subsets and the pDC2 : pDC1 ratio showed good intra-and interassay reproducibility. Compared with patients in group C, those in groups A and B demonstrated a significantly higher relative incidence of pDC2 and a lower incidence of pDC1 - similar to those values observed in normal healthy controls. Moreover, the pDC2 : pDC1 ratio was significantly higher in patients undergoing (successful) weaning and in those off immunosuppression compared with patients on maintenance immunosuppression. PMID- 12780561 TI - Preservation of pancreas tissue during cold storage assessed by X-ray microanalysis. AB - Clinical transplantation requires cold storage of tissue for several hours. We have examined the elemental content in exocrine and endocrine cells in mouse pancreas after cold storage by X-ray microanalysis, and in parallel carried out morphological studies. Tissue was stored at 4 degrees C for 4-12 h in Normal Krebs-Ringer's (high Na+/K+ ratio), Modified Krebs-Ringer's (low Na+/K+ ratio), Euro-Collins, University of Wisconsin (UW) solution, and seven modified version of UW solution. Incubation in Normal Krebs-Ringer's solution caused significantly increased Na and decreased K concentrations in contrast to incubation in other solutions. The cellular concentration of Na and Cl followed the concentration in the storage solution. Changes in the endocrine cells were similar to, but less pronounced than those in exocrine cells. Calcium was retained best in UW and some variants of UW, and least in Euro-Collins. This may indicate differences in preservation of secretory granules. Also, morphological studies showed that endocrine cells were less affected than exocrine cells. In conclusion, the only factor determining the intracellular concentration of diffusible ions after cold tissue storage is the ionic composition of the extracellular medium. X-ray microanalysis provides an objective method to assess whether the intracellular ionic composition of tissue is maintained during storage. PMID- 12780562 TI - Antibody-mediated rejection criteria - an addition to the Banff 97 classification of renal allograft rejection. AB - Antibody-mediated rejection (AbAR) is increasingly recognized in the renal allograft population, and successful therapeutic regimens have been developed to prevent and treat AbAR, enabling excellent outcomes even in patients highly sensitized to the donor prior to transplant. It has become critical to develop standardized criteria for the pathological diagnosis of AbAR. This article presents international consensus criteria for and classification of AbAR developed based on discussions held at the Sixth Banff Conference on Allograft Pathology in 2001. This classification represents a working formulation, to be revisited as additional data accumulate in this important area of renal transplantation. PMID- 12780563 TI - Improved scoring system to assess adult donors for cadaver renal transplantation. AB - We previously proposed a quantitative approach to assess donor organs for cadaver renal transplantation. To improve on our original scoring system, we studied 34 324 patients who received cadaver renal transplants from adult donors between 1994 and 1999 and were reported to the UNOS Scientific Renal Transplant Registry. A scoring system was developed from five donor variables (age, 0-25 points; history of hypertension, 0-4; creatinine clearance before procurement, 0-4; cause of death, 0-3; HLA mismatch, 0-3) that showed a significant correlation with renal function and long-term graft survival. Cadaver kidneys were stratified by cumulative donor score: grade A, 0-9 points; grade B, 10-19; grade C, 20-29; and grade D, 30-39. The influence of donor score on renal function and graft survival was most severe above 20 points, designated 'marginal' kidneys. In summary, a donor scoring system developed from a large population database was useful in predicting outcome after cadaver renal transplantation. The improved system provides a quantitative approach to evaluation of marginal kidneys and may improve allocation of these organs in cadaver renal transplantation. PMID- 12780565 TI - Cytomegalovirus disease in high-risk transplant recipients despite ganciclovir or valganciclovir prophylaxis. AB - The clinical patterns and predictors of cytomegalovirus (CMV) disease in kidney and/or pancreas transplant patients on ganciclovir (1.0 g po t.i.d.) or valganciclovir (450 mg po q.d.) prophylaxis were studied. This is a retrospective analysis of 129 transplant recipients. Median follow up was 12 months (range, 6 18 months). The overall incidence of CMV disease at 1-year post-transplant was 14% (4% tissue-invasive, 10% noninvasive). Seventeen of 18 patients were diagnosed with CMV after completion of 3 months' prophylaxis (median 8 weeks, range, 2-28 weeks). Induction treatment with thymoglobulin, and Donor +/Recipient - CMV status were the strongest predictors for the development of CMV disease. Cytomegalovirus incidence was not different between patients treated with ganciclovir or valganciclovir (15 vs. 17%, respectively). Valganciclovir (450 mg q.d.) is as effective as oral ganciclovir in CMV prophylaxis. High-risk individuals might require higher doses or longer duration of valganciclovir treatment. PMID- 12780564 TI - Campath-1H induction plus rapamycin monotherapy for renal transplantation: results of a pilot study. AB - Campath-1H, an anti-CD52 monoclonal antibody, was used as induction therapy (40 mg i.v. total dose) in 29 primary human renal transplants, and the patients were maintained on rapamycin monotherapy (levels 8-15 ng/mL) post-transplant. Campath 1H profoundly depletes lymphocytes long-term and more transiently depletes B cells and monocytes. All patients are alive and well at 3-29 months of follow up. One graft was lost because of rejection. There have been no systemic infections and no malignancies. Eight of 29 patients have experienced rejection, which was successfully treated in seven of eight patients. Five of these patients had pathological evidence of a humoral component of their rejection. Seven of the 29 patients were converted to standard triple therapy on account of rejection. Rapamycin was generally well tolerated in that there were no significant wound healing problems; two lymphoceles required surgical drainage; and most patients were treated with a lipid-lowering agent. Flow crossmatch testing post-transplant revealed evidence of alloantibody in two patients tested with previous combined cellular and humoral rejection. Biopsies have shown no chronic allograft nephropathy to date. In view of the relatively high incidence of early humoral rejection, we plan to modify the immunosuppressive regimen in subsequent pilot studies. This clinical trial provides insight into the use of Campath-1H induction in combination with rapamycin maintenance monotherapy. PMID- 12780566 TI - A role for indirect allorecognition in lung transplant recipients with obliterative bronchiolitis. AB - Obliterative bronchiolitis (OB) occurs in 50% of patients 2 years after lung transplantation. Although a correlation between OB and indirect recognition of donor peptides has been reported, the relative roles of direct vs. indirect recognition have not been investigated. Limiting dilution analysis was used to determine the frequencies of recipient T-helper cells recognizing donor and third party alloantigens in 19 patients (8 OB positive, 11 OB negative). The assays were designed to distinguish between indirect and direct presentation. In three patients the direct and indirect assay were performed on the same blood sample. Six out of seven patients with OB were hyperresponsive in the indirect pathway to donor antigens compared to third-party, the corresponding figure for OB negative patients being 2/7. In contrast, 5/7 patients were hyporesponsive in the direct pathway; hyporesponsiveness in the direct pathway did not correlate with freedom from OB. The patients in whom the assays were performed from the same blood sample confirmed that donor specific hyperresponsiveness in the indirect route can coexist with donor-specific hyporesponsiveness in the direct route. In conclusion, lung allograft recipients, like recipients of other organ allografts, become hyporesponsive in the direct route but sensitization via the indirect pathway is associated with chronic rejection. PMID- 12780567 TI - Characterization of hepatitis B virus surface antigen and polymerase mutations in liver transplant recipients pre- and post-transplant. AB - We evaluated serum samples from 18 chronic hepatitis B virus (HBV) patients who underwent liver transplantation for the presence of HBV polymerase and S gene mutations and HBV genotype using a new commercially available sequencing assay. All three patients with hepatitis B immune globulin (HBIG) treatment failure followed by nucleoside analogue treatment failure were infected with HBV genotype C; a pre-existing HBV S antigen (HBsAg) mutation (sD144A) was identified in one patient pretransplant, while sG145R mutations emerged in the other two patients post-transplant. These HBsAg mutations persisted for the duration of the study (5 6 years), despite the absence of HBIG administration for a 4-5-year period. Significant viral polymerase mutations (rtL180M and rtM204I/V) also emerged in all of these patients following treatment with lamivudine and/or famciclovir. Four of six patients with HBIG breakthrough without nucleoside analogue treatment failure yielded potentially significant HBsAg mutations post transplant. These data do not support previous reports highlighting the disappearance of HBsAg mutants in liver transplant recipients after discontinuation of HBIG. Determination of HBV genotype, as well as identification of HBV polymerase and S gene mutations in liver transplant candidates may be warranted to optimize HBV management strategies post transplant. PMID- 12780568 TI - CD32-mediated platelet aggregation in vitro by anti-thymocyte globulin: implication of therapy-induced in vivo thrombocytopenia. AB - Induction therapy with polyclonal antithymocyte-globulin (ATG) is widely used in the prophylaxis and treatment of acute cardiac-allograft rejection. Thrombocytopenia, however, is a major side-effect of ATG therapy and its mechanisms are poorly understood. The influence of ATG on platelet aggregation was studied aggregometrically, expression of platelet surface activation antigens CD62P and CD63 was determined by flow cytometry analysis, and electron microscopy was utilized to determine thrombocyte morphology. Treatment of platelets with ATG markedly induced aggregation, whereas OKT3 or anti-IL-2R antibodies did not. Furthermore, platelets incubated with ATG featured an up-regulation of the surface activation markers CD62P and CD63, secretion of platelet-bound sCD40L (CD154) and increased signs of aggregation in electron microscopy analysis. The capacity of ATG to induce platelet aggregation was completely blocked by antibodies against the low-affinity Fc IgG receptor (CD32). Since blocking of CD32 abrogates platelet aggregation, we suggest that CD32 plays a crucial role in ATG-induced thrombocytopenia. PMID- 12780569 TI - Veno-atrial bypass for the operative treatment of septic gas gangrene secondary to delayed hepatic artery thrombosis. AB - The occurrence of late hepatic artery thrombosis after orthotopic liver transplantation can result in gas gangrene of the graft. This clinical scenario has the potential to be rapidly fatal as a result of fulminant hepatic failure, sepsis and multiple-organ-failure syndrome. Emergency operative intervention is indicated to remove the septic source and replace the failed liver. In this report, both cases demonstrated rapid deterioration within 24 h from the onset of symptoms, in spite of maximum supportive care. Intra-operative handling of the gangrenous graft resulted in hemodynamic instability and a technically unfeasible hepatectomy. The use of extra-corporeal veno-atrial bypass, by isolating the septic source, allowed for graft hepatectomy and successful re-transplantation in the second of these reported cases. PMID- 12780571 TI - Rectal Cancer: Where are we now? PMID- 12780570 TI - Calicivirus enteritis in an intestinal transplant recipient. AB - Protracted diarrhea of uncertain etiology is a significant problem following intestinal transplantation. We report an infant who developed severe secretory diarrhea 178 days after intestinal transplantation that persisted for more than 120 days. Repeated allograft biopsies demonstrated only nonspecific inflammation. Enzyme immunoassay (for rotavirus), culture, and reverse transcription polymerase chain reaction [calicivirus (Norwalk-like virus)] were used to identify the allograft viral infection. A heavy density of calicivirus RNA nucleotide sequences (genogroup II, strain Miami Beach) was isolated from the jejunal and ileal allograft. Following a reduction in immunosuppressive therapy, diarrhea and enteritis remitted in association with the disappearance of all calicivirus RNA sequences. Calicivirus may cause severe allograft dysfunction in intestinal transplant recipients. PMID- 12780572 TI - The colonic J-pouch in colo-anal anastomosis. AB - Anastomosis of the colon to the anal canal is now an accepted technique in the surgical management of low and mid rectal cancers. Although significant postoperative bowel disturbance is often seen with straight colo-anal anastomosis, controversy exists over the benefit of adding a colonic pouch for low anastomoses. Several short and long-term studies have demonstrated the early functional superiority of pouch-anal over straight anastomosis. Pouch construction does not compromise anal physiological parameters. It is recommended the pouch be constructed from a length of descending colon and be small (5 cm) in size to adequately act as a neo-rectum; long-term evacuatory difficulties are encountered with the construction of large pouches (10 cm). Anastomotic complications appear to be less frequent with pouch surgery; construction of a pouch does not significantly add to operative time, patient morbidity and mortality. At present there is no compromise to long-term oncological survival. The data supporting these statements is weak and based largely upon retrospective studies. Furthermore the impact of improved function with pouch-anal anastomosis on overall quality of life has been poorly investigated. Further prospective randomized studies are required to ascertain whether the potential benefits of a colonic pouch are realized in the randomized setting. PMID- 12780573 TI - Spotlight on a distant mirror - colorectal cancer surgery in a 20-year period at a general hospital. AB - BACKGROUND: A published audit of the management of colorectal cancer at a general hospital in the 1970s was available for comparison with a later audit at the same hospital in the 1990s. METHODS: Case note analysis. RESULTS: In the later audit, more cases were treated annually by an unchanged surgical team. The incidence of synchronous combined excision of the rectum, for rectal cancers suitable for resection, was halved, and that of anterior resection of the rectum (sphincter sparing, without a permanent stoma) increased almost threefold. The incidence of local recurrence in cases suitable for rectal surgery dropped from 17% to 9%, in spite of the change in the principal operation undertaken for this population. Outcomes associated with critical care improved as resources in this discipline became available. Overall survival figures were only improved by 6% in the20-year period, reflecting a diagnosis of Dukes C tumours or worse in at least 45% of the stable population studied in both audits. CONCLUSION: More resources are necessary in Great Britain to increase survival figures in this common cancer. Earlier diagnosis and more specialist management of the disease may allow us to emulate American and Swedish survival figures. PMID- 12780574 TI - Topical 0.2% glyceryl trinitrate ointment for anal fissures: long-term efficacy in routine clinical practice. AB - OBJECTIVE: Randomized controlled trials have reported fissure healing rates of 60 70% using topical 0.2% GTN ointment, but the effectiveness of this therapy in routine clinical practice, particularly in the long term, is uncertain. This study aimed to evaluate the efficacy of topical GTN for anal fissures in an outpatient setting. PATIENTS AND METHODS: A case note review and postal questionnaire survey were undertaken for patients with a diagnosis of anal fissure dispensed 0.2% GTN ointment from one hospital pharmacy over a two year period (June 1996-May 1998). RESULTS: Ninety-three patients (53 male) applied 0.2% GTN ointment twice daily to the anoderm for anal fissure. They were reviewed on average 8 (2-16), weeks later, by which time 57 (61%) fissures had healed, 33 persisted and 3 patients were lost to follow up. Seventy-two patients (41male), median age 42 (22-83) years, returned completed questionnaires (77% response rate). Forty-nine (68%) had healed with GTN, but 25 had recurrent symptoms after a median of 6 (1-18) months. Sixteen of those patients reporting symptomatic recurrence were prescribed further GTN for a recurrent fissure: 14 (88%) healed, but 2 persisted, and had surgery. In the other nine patients symptoms resolved spontaneously. Thirty-five (49%) experienced headaches, 3 (4%) discontinuing treatment as a result. The median follow up was 25 (13-36) months. CONCLUSION: 0.2% GTN heals 60% of fissures in the outpatient setting but half the patients develop headaches. Up to one third of healed fissures may recur within 18 months but the majority respond to further GTN. Only 4% of patients initially healed with GTN later require sphincterotomy for recurrences. PMID- 12780575 TI - Treatment of enterocele by abdominal colporectosacropexy - efficacy on pelvic pressure. AB - OBJECTIVE: Enterocele induces pelvic pressure, obstructed defaecation, lower abdominal pain and/or false urge to defaecate in patients. The aim of this study was to evaluate the efficacy of abdominal colporectosacropexy in these symptoms, especially on pelvic pressure. METHODS: Sixty-two consecutive women with enterocele were included. All patients were symptomatic because they had: pelvic pressure (n = 62), obstructed defaecation (n = 40), lower abdominal pain (n = 8) or faecal incontinence (n = 16). Defaecography confirmed enterocele in all patients. The surgical procedure was performed by the same surgeon and was an abdominal colporectosacropexy with a nonabsorbable Prolene(R) mesh. After surgery, clinical evaluation (62/62 patients) and a telephone questionnaire (56/62 patients) were performed, respectively, 3 months and 27 +/- 13 months after surgery. RESULTS: Defaecography showed rectal abnormalities associated with enterocele in 59/62 patients (rectocele, rectal prolapse). No recurrence of enterocele was observed 3 months after surgery, but 1 patient demonstrated recurrence 10 months after surgery. Pelvic pressure was less frequent after abdominal colporectosacropexy, than before surgery (P < 0.01): pelvic pressure totally disappeared in 41/56 patients, and partially in 10/56 patients. The number of patients with obstructed defaecation, lower abdominal pain, or faecal incontinence was not different before and 27 months after surgery. The number of patients with urinary incontinence was also not different before and after surgery (30 and 27 patients). CONCLUSIONS: This study of a large number of patients with enterocele shows that abdominal colporectosacropexy improves pelvic pressure in most patients and does not modify urinary status. PMID- 12780576 TI - Scintigraphic assessment of colostomy irrigation. AB - OBJECTIVE: This study aims to evaluate colonic transport following colostomy irrigation with a new scintigraphic technique. MATERIALS AND METHODS: To label the bowel contents 19 patients (11 uncomplicated colostomy irrigation, 8 complicated colostomy irrigation) took 111In-labelled polystyrene pellets one and two days before investigation. 99mTc-DTPA was mixed with the irrigation fluid to assess its extent within the bowel. Scintigraphy was performed before and after a standardized washout procedure. The colon was divided into three segments 1: the caecum andascending colon; 2: the transverse colon; 3: the descending and sigmoid colon. Assuming ordered evacuation of the colon, the contribution of each colonic segment to the total evacuation was expressed as a percentage of the original segmental counts. These were added to reach a total defaecation score (range: 0 300). RESULTS: In uncomplicated colostomy irrigation, the median defaecation score was 235 (range: 145-289) corresponding to complete evacuation of the descending and transverse colon and 35% evacuation of the caecum/ascending colon. In complicated colostomy irrigation it was possible to distinguish specific emptying patterns. The retained irrigation fluid reached the caecum in all but one patient. CONCLUSION: Scintigraphy can be used to evaluate colonic emptying following colostomy irrigation. PMID- 12780577 TI - The influence of a colorectal service on the outcome of patients with colorectal cancer. AB - OBJECTIVE: To perform a prospective audit of outcomes and survival of all patients presenting to a colorectal service with colorectal cancer, and to compare these results with an historical control group. PATIENTS AND METHODS: At a community based teaching hospital, a prospective audit of outcomes and survival of patients with colorectal cancer was compared with a historical control. The study included all patients referred to a colorectal service with colorectal cancer from 1996 to 2000 (5-year period). The control group was a retrospective review of patients presenting to the same hospital with colorectal cancer from 1989 to 1994 (6-year period). A Kaplan-Meier survival analysis compared the overall survival (all-cause mortality) between the two groups. RESULTS: When comparing the study periods 1989-95 (n = 477) to 1996-2000 (n = 323), there has been a significant reduction in postoperative stay (16.2 vs 8.0 days, P < 0.05), and a reduction in postoperative mortality (4.5%vs 2.7%, n.s.). There was a significant increase in the overall 2 years survival for patients with colorectal cancer (62% to 71%, P < 0.01). There was also a significant increase in the overall 2 years survival of patients with rectal cancer (66% to 74%, P < 0.01), patients with ACPS C colon cancers (64% to 83%, P < 0.05), and ACPS C rectal cancers (74% to 85%, P < 0.01). CONCLUSIONS: There have been significant gains in the survival of patients presenting to a community based teaching hospital with colorectal cancer. These improvements have been most notable in patients with nodal metastases at the time of diagnosis. PMID- 12780578 TI - Total anorectal reconstruction with an artificial bowel sphincter: Report of five cases with a minimum follow-up of 6 months. AB - BACKGROUND: The artificial bowel sphincter (Acticon ABS - American Medical Systems, Minneapolis, MN, USA) has been proposed as a treatment for patients with faecal incontinence. The good results achieved with this procedure encouraged us to utilize this device for reconstruction of patients who previously underwent an abdominoperineal resection (APR). METHOD: Between 1999 and 2000 we implanted the ABS in five patients undergoing an APR. One patient was male and four female, the mean age was 51.3 years. Three patients had been operated on for rectal cancer, one for rectal agenesia and one for a giant benign tumour of the pelvis. RESULTS: The length of follow up ranged from 6 to 22 months. Manometry assessed a basal pressure with the ABS cuff inflated between 58 and 62.2 mmHg. All but one achieved a good grade of continence with a Wexner score range between 3 and 9. A certain degree of impaired evacuation occurred in two patients but, with adequate training, this improved and did not affect patient satisfaction. CONCLUSION: The ABS is a good option for reconstruction of patients previously treated with an APR. As compared to electrostimulated graciloplasty the ABS technique seems to be easier to perform and more acceptable for the patients, although the cost of the device is still high. PMID- 12780579 TI - Severe dysplastic lesions in the colon - how aggressive should we be? AB - INTRODUCTION: Biopsies of colonic lesions are often reported as showing dysplasia, though in reality some lesions may harbour invasive malignancy. AIM: To assess the risk of underlying invasive malignancy in sessile polyps where biopsies had shown severe dysplasia and also to attempt to define a management strategy in such patients. METHODS: Between 1997 and 2001, 30 patients were diagnosed as having severe dysplasia using Morson's criteria in colonic lesions not amenable for endoscopic polypectomy. Severely dysplastic lesions were completely excised by appropriate surgical measures. RESULTS: Out of 30 patients, 15 had invasive cancers. Surgical intervention involved anterior resections, endoanal excisions, sigmoid colectomies, or abdomino-perineal excisions as deemed appropriate. The lesions ranged in size from 0.5 cm to 13 cm (mean 3.4 cm). There were nine T1 lesions (one of which was T1N1) and two each of T2, T3, T4 lesions (10 Dukes' A, 3 Dukes' B, 2 Dukes' C). Complete resection was confirmed histologically in all cases. One patient had a leak following endoanal excision, which required intervention. There was no mortality. DISCUSSION: This study demonstrates that endoscopic sampling can be misleading and severely dysplastic sessile lesions should be managed along the same principles as followed for invasive cancers, rather than adopting a 'wait and watch' policy with repeated endoscopies, biopsies or piece-meal polypectomies. PMID- 12780580 TI - Variability in serotonin and enterochromaffin cells in patients with colonic inertia and idiopathic diarrhoea as compared to normal controls. AB - AIM: To evaluate differences in distribution, density and staining intensity of enterochromaffin cells (EC) and serotonin cells (SC) in the colonic mucosa of patients with colonic inertia (CI), idiopathic diarrhoea (ID) and a control group. METHODS: Three groups were studied: 19 patients' colons after subtotal colectomy for CI, and 17 patients' biopsies for diarrhoea (>3 bowel movements/day) with histological findings of normal mucosa (excluding microscopic, eosinophillic and collagenous colitis). The third group included 15 patients who underwent colonoscopy and biopsy for indications other than constipation, inflammatory bowel disease, diarrhoea or neoplasm (control group). Specimen blocks were obtained in each case from the right and left colon. Immunohistochemical staining for EC and SC were done on 4 micro m sections from Hollandes fixed, paraffin embedded tissues with primary rabbit antibody against chromagranin A or serotonin, and biotynylated secondary antibody and enzyme labelled streptavidin. RESULTS: The number of EC in the mucosa of the left colon in patients with CI (16.8 +/- 10.2) and ID (19.9 +/- 9.7) were significantly higher than they were on the right side (CI: 9.4 +/- 6.0, ID: 12.1 +/- 5.3). However, there were no significant differences between the left and right sides in the control group (L: 10.3 +/- 5.3; R: 13.4 +/- 7.6). Although the quantity of EC in the left colon in both patients with CI (P < 0.05) and ID (P < 0.01) were significantly higher than in the controls, there was no significant difference between CI and ID. In both the right and left colon, the percentage of EC with low positive density was significantly higher (P < 0.01) while those cells with moderate or low staining intensity were significantly lower in patients with CI than in either patients with ID or control group. In patients with CI, the quantity of SC in the mucosa of the left colon (12.1 +/- 6.4) was higher than in the right (CI: 7.9 +/- 3.6; control 4.6 +/- 3.3; ID 4.6 +/- 2.9) (P = 0.0057). In contrast there was no significant difference in SC in either the ID or control groups. The quantity of SC in both sides of the colon was significantly higher both in patients with CI as compared to the control group (P < 0.01) and patients with CI vs. patients with ID (L = P < 0.01; R = P < 0.05). There was a significantly positive correlation between the numbers of EC and SC in patients with CI (L: r = 0.5425, P < 0.05; R: r = 0.745, P < 0.01). CONCLUSION: In patients with CI, EC increases possibly due to an increase in SC. Conversely, in patients with ID, the EC increase results from peptides other than SC. Our results suggest that different aetiological factors contribute to ID and CI. PMID- 12780581 TI - Demonstrating the clinical and cost effectiveness of adhesion reduction strategies. AB - OBJECTIVE: To examine the feasibility of conducting Randomized Controlled Trials (RCT) in lower abdominal surgery to demonstrate a reduction in adhesion-related admissions following use of an adhesion reduction product, and to model the cost effectiveness of such products. METHODS: The number of patients in each limb of a RCT comparing an adhesion reduction product to a control has been estimated based on 25% and 50% reductions in adhesion-related readmissions one year after surgery, for P = 0.05 at a power of 80% and P=0.01 at a power of 90%. A cost effectiveness model based on the Surgical and Clinical Adhesions Research Group (SCAR) database has been developed which calculates the percentage reduction in readmissions required of an adhesion reduction product to return the cost of investment. It also estimates the cumulative costs of adhesion-related readmissions for lower abdominal surgery and the cost savings associated with an adhesion reduction policy using a low or high cost product. RESULTS: 7.2% of patients undergoing lower abdominal surgery will readmit due to adhesions in the first year after surgery. To demonstrate a 25% reduction in readmissions one year after surgery, it is calculated that a RCT would require between 5686 (P = 0.05, power=80%) and 7766 (P = 0.01, power = 90%) lower abdominal surgery patients followed-up for one year. A cost effectiveness analysis demonstrates that routine use of adhesion reduction products costing pound 50 per patient will payback the cost of such investment if they reduce adhesion-related readmissions by 16% after 3 years. A product costing pound 200 will need to offer a 64.1% reduction in readmissions after 3 years. For the estimated 158 000 lower abdominal surgery operations conducted in the UK each year, the cumulative costs of adhesion related readmissions over 10 years are estimated at pound 569 Million. CONCLUSION: Demonstrating the clinical effectiveness of adhesion reduction products in the RCT setting is unlikely to be feasible due to the large number of patients required. Products costing pound 200 or more are unlikely to payback their direct costs. PMID- 12780583 TI - The Cancer Collaborative Services Project. PMID- 12780582 TI - Safety of the temporary loop ileostomy. AB - OBJECTIVE: To evaluate the complications of the temporary loop ileostomy. METHOD: A retrospective study of 222 consecutive patients with low anterior resection, ileal pouch-anal anastomosis or continent ileostomy and a diverting loop ileostomy routinely fashioned during the primary operation. The loop ileostomy was closed in 213 patients (96%) during the minimum follow-up period of 15 months. RESULTS: Four patients (2%) required preterm closure of the ostomy due to stomal retraction (n = 3) or bowel obstruction (n = 1). Four patients were readmitted due to transient bowel obstruction that resolved without surgery. After closure of the loop ileostomy a total of 27 patients (13%) had complications. In 7 patients emergency re-operation was done due to small bowel obstruction (n = 5) or intra-abdominal abscess (n = 2). Elective re-operation was done in 5 patients for hernia at the site of the previous stoma. Despite the use of a loop ileostomy there was 1 postoperative death after the initial operation in consequence of anastomotic leakage. There was 1 death in consequence of closure of the loop ileostomy after 3 weeks due to intra-abdominal sepsis and heart failure. CONCLUSION: In this series closure of the ostomy wasassociated with one death (0.5%) and overall ostomy-related morbidity included the need to re-operate in 6%. PMID- 12780584 TI - Acute rectal obstruction after PPH stapled haemorrhoidectomy. AB - An unusual case of acute intestinal obstruction following a PPH stapled haemorrhoidectomy is reported: the complication was due to an almost complete interruption of the rectal lumen tightened by a purse-string suture anchored by staples. The stricture was dilated and the suture released thus restoring intestinal continuity. Surgeons should be aware that such troublesome complication may occur and can be treated successfully by a transanal approach. PMID- 12780585 TI - Anal canal metastases from left sided colorectal cancer. PMID- 12780587 TI - European Association of Coloproctology Annual Meeting, Maastricht, 14-15 September 2001. PMID- 12780586 TI - European Association of Coloproctology Second Annual Scientific Meeting, 13-15 September, 2001, Maastricht, The Netherlands. PMID- 12780590 TI - ACPGBI: the 12th Annual Meeting. PMID- 12780591 TI - The management of persistent and recurrent chronic anal fissures. AB - There are numerous definitive reviews concerning aetiology and management of acute and chronic anal fissures. The problem of persistence and recurrence after surgical and nonsurgical therapy has not been specifically addressed and there is little evidence-based guidance for the management of this difficult group of patients. A review of the literature with particular reference to persistence and recurrence of chronic anal fissures is presented and an algorithm of management incorporating evidence-based data is suggested. PMID- 12780592 TI - Does perifascial rectal excision (i.e. TME) when combined with the autonomic nerve-sparing technique interfere with operative radicality? AB - OBJECTIVE: The lymphatic drainage from the rectum was studied to evaluate if the autonomic nerve sparing dissection may interfere with the operative radicality and might result in metastatic lymph nodes being overlooked and left in situ. PATIENTS AND METHODS: 50 consecutive patients had an extended extrafascial rectal excision resection for cancer. In 19 of the 50 patients activated carbon particles (CH40) were injected preoperatively into the rectum. The autonomic nerves with surrounding connective tissue were serially dissected from the resected specimen, carefully sliced at 5-mm intervals and collected for histological study. Lymph nodes along the axial and lateral drainage routes were examined, and the inclusion of CH40 in the nodes was microscopically studied according to the site of CH40 injection. RESULTS: Lymph nodes within the connective tissue along the dissected autonomic nerves were demonstrated in 47 of the 50 cases. Two of 50 cases had positive nodes along preaortic plexus or pelvic plexus, and a case with nodal involvement along the pelvic plexus had poor prognosis in spite of nerve excision. CH40 when injected into the rectum above the peritoneal reflection was demonstrated in the vast majority of the axial nodes, while in only one lymph node along the preaortic plexus when injected in the rectum above the peritoneal reflection. On the other hand when injected in the rectum below the peritoneal reflection, CH40 was demonstrated both in axial and lateral nodes as well as in lymph nodes along bilateral pelvic plexuses, right hypogastric nerve, superior hypogastric plexus, preaortic plexus and mesenteric plexus as well. CONCLUSIONS: When located above the peritoneal reflection a rectal carcinoma will spread preferentially along the upper axial route, while a carcinoma located below the peritoneal reflection will also spread laterally and along the autonomic nerves. It was inferred that lymphatic flow along the autonomic nerves came up from the rectum below the peritoneal reflection mainly through a so-called lateral ligament but its clinical significance was negligible. Therefore doing TME with autonomic nerve preservation does not imply a less radical surgery from the point of lymphatic spread. PMID- 12780593 TI - Palliative stenting of malignant large bowel obstruction. AB - OBJECTIVE: Untreated malignant large bowel obstruction is rapidly fatal. Short term palliation of symptoms can be achieved by formation of a stoma in those patients for whom resection surgery is inappropriate. In the final months of life, a stoma represents a significant burden for both patients and carers. Palliative endoluminal stenting may therefore be an attractive alternative option for this poor prognosis group. In this paper, we examine our experience of palliative endoluminal colonic stenting. PATIENTS: Twenty patients, 11 males and 9 females of median age 81 years were referred for stenting. All had left sided colonic cancers. Ten patients had confirmed metastases on presentation, four had fixed rectal cancers and the remainder had severe comorbidity limiting surgical options. Stents were placed endoscopically using a radiologically controlled 'stent over wire' technique. RESULTS: Stenting successfully relieved the obstruction in 18 of the 20 patients attempted. In one patient the stricture could not be negotiated and the procedure was abandoned. Eleven patients have died of their disease, their median duration of palliation was 50 days (3-152 days). The rest of the patients continue in follow-up and have had 80 days median palliation (14-257 days). One stent-related complication has been observed in a patient who suffered anal pain due to fracture and migration of part of a stent into the low rectum. This complication occurred after 250 days and the distal stent fragment was removed with further symptom relief. CONCLUSION: Carefully selected patients benefit from colonic endoluminal stenting with relief of obstructive symptoms. They may be spared the potential problems associated with palliative stoma formation. PMID- 12780594 TI - Management of recurrent colorectal carcinoma. AB - BACKGROUND/OBJECTIVE: The records of patients treated for adenocarcinoma of the colon and rectum between 1 January 1988 and 31 December 1995 at Naval Medical Center San Diego were reviewed. Analysis was made of patients who developed recurrences after potentially definitive primary therapy. METHODS: A retrospective review of 410 patients diagnosed with colorectal cancer at our institution was conducted. The focus of this review was to identify patients with recurrent disease after curative initial procedures, and to determine how recurrences were detected and treated. Survival data for 48 patients undergoing various curative and palliative procedures, or no therapy, were generated. RESULTS: The decision to re-operate with curative intent was made after a multidisciplinary review of restaging studies. Laparoscopy was not used in this determination. Curative resection of recurrence confers increased survival over non-curative surgery and no surgery (P < 0.001). This is misleading because of patient selection; several patients undergo potentially curative surgery but are determined intraoperatively to best be palliated, or to have further surgery aborted. Analysis of results in patients undergoing potentially curative surgery vs. those undergoing planned palliation vs. those not operated reveals that these also provide significantly different outcomes (P < 0.003). CONCLUSIONS: Proper delineation of resectable lesions in patients with recurrent colorectal cancer contributes to better outcomes for them. That determination is difficult, and efforts are underway in our institution and elsewhere to better delineate which patients are optimal preoperatively. We consider multidisciplinary Tumor Board evaluation to be central to this process. PMID- 12780595 TI - The outcome of surgery for complex anal fistula. AB - OBJECTIVE: The aim of this study was to examine the results of surgery for complex anal fistulas treated by a variety of techniques, in terms of fistula healing, recurrent anal sepsis and effect of surgery on anal continence. PATIENTS AND METHODS: This study included 63 patients with complex fistulas treated between November 1995 and September 1999. A variety of techniques were employed, including short-term loose seton drain (12), long-term loose seton drain (11), cutting seton (17), and rectal advancement flap (19). Outcome was assessed at clinic review and continence was further assessed by detailed questionnaire sent to the patients sometime after surgery. RESULTS: Healing occurred in 9 (75%) patients treated with a short-term, loose drainage seton; 16 (94%) patients treated with a cutting seton and 17 (89%) patients in the rectal advancement flap group. Incontinence reported at clinic review seemed to be more frequent in the advancement flap group. However, a detailed continence questionnaire revealed that 50% of patients reported episodes of incontinence to flatus or liquid after all techniques, which had not been detected at routine clinical review. Incontinence to solids was only reported by two of the patients who had been treated with a cutting seton. CONCLUSIONS: Complex fistulas may be successfully treated by a variety of techniques. Disturbed anal continence following surgery is common and worse than clinic assessment would suggest. PMID- 12780596 TI - Simple equipment for decompression of the colon during laparotomy for large bowel obstruction. AB - A simple method is described for decompressing the colon at laparotomy for large bowel obstruction. PMID- 12780597 TI - Femoral nerve injury as a complication of pelvic surgery. AB - Femoral nerve injury is a rare complication of pelvic surgery but may lead to medicolegal actions. The common cause is the use of a long-bladed self-retaining pelvic retractor. The injury is not well recognised in coloproctology practice and this brief article is to draw attention to this complication. PMID- 12780598 TI - Long-term results of modified graciloplasty for sphincter replacement after rectal excision. AB - OBJECTIVE: Restoration of the anal sphincter by means of electrically stimulated (dynamic) graciloplasty is a new therapeutic option for patients with severe faecal incontinence or those having abdomino-perineal resection (APR) of the anorectum. The present study reviews the outcome of total anorectal reconstruction (TAR) after APR for low rectal cancer or recurrent anal cancer. METHODS: From 1992 to 2000, 35 of 64 patients treated with dynamic graciloplasty had a TAR performed either synchronously (n=26) or as a secondary procedure one to five years after rectal excision (n = 9). RESULTS: The most frequent complication was injury or erosion of the neorectum (n = 9) which, was avoided with increasing surgical experience. Defaecation disorders and consequent incontinence were the most common functional problem and had to be treated with periodical enemas. CONCLUSION: Although sphincter replacement by means of TAR after APR led to poorer functional results than those achieved in patients treated with dynamic graciloplasty for faecal incontinence, TAR remains a valid treatment option for patients who do not tolerate a permanent stoma. PMID- 12780599 TI - Colonic resection for colovesical fistula: 5-year follow-up. AB - OBJECTIVES: The outcome of colovesical fistula management may be unsatisfactory; complications are reported in up to 45% of patients. Published studies are retrospective and tend to lack standardized management strategies and long-term follow-up. This cohort study assesses a policy of resection of colovesical fistulae in continuity with any distal colorectal stricture, and includes 5-year follow-up. METHOD: All patients undergoing surgery in our institution for colovesical fistula between February 1991 and April 1995 were entered into the study. The fistulae were resected in continuity with any distal bowel stricture, according to a standard single-stage operative protocol. Postoperative mortality and morbidity were recorded, and prospective review was undertaken at April 2000. RESULTS: Nineteen consecutive patients entered the study. The source of the fistula was diverticular disease (n = 14), colorectal cancer (n = 3), trauma (n = 1) or Crohn's (n = 1) disease. Thirteen patients had a colorectal stricture. One patient died due to ischaemic colitis within 30 days of surgery. Eleven other patients died of unrelated causes before April 2000, in whom there was no evidence of fistula recurrence before death at a median of 37 months after operation (range 2-95 months). At 5-year follow-up there was no evidence of fistula recurrence in the seven remaining patients. CONCLUSIONS: A policy of resection of the fistula and associated colorectal stricture with primary bowel anastomosis and bladder drainage, resulted in no recurrences and low morbidity. However comorbidity is important in this patient population, most of whom will die from unrelated causes within a few years. PMID- 12780600 TI - Overlapping sphincteroplasty: does preservation of the scar influence immediate outcome? AB - BACKGROUND: The importance of the overlapping scar in an anterior sphincteroplasty is often emphasized. The aim of this study was to identify the tissue type used in overlapping sphincter repair based upon ultrasound images, and to correlate these results with the immediate clinical outcome. METHODS: Data were collected prospectively on all patients with faecal incontinence who underwent anterior overlapping sphincteroplasty between June 1998 and May 1999. Continence was assessed by a standardized incontinence score ranging from 0 to 20. Pre-operative ultrasound images were compared to intraoperative ultrasound findings for each patient. In each case the surgeon performed an overlap of what was grossly felt to represent scar after which a single blinded observer performed intraoperative ultrasound. The degree of overlap was measured and classified as hyperechoic over hyperechoic (muscle over muscle; Type 1), hyperechoic over or under hypoechoic (muscle over or under scar; Type 2), hypoechoic over hypoechoic (scar over scar; Type 3). The patient follow-up included incontinence score that was obtained by telephone interview; suboptimal outcome was considered as an incontinence score >/= 6. Statistical analysis was performed using the Mann-Whitney test and Wilcoxon matched-pairs test. RESULTS: Fourteen female patients with a mean age of 51.6 (range 28-79) years were evaluated. The mean pre-operative incontinence score was 17.1 (range 7-20) and 13 of the 14 (93%) patients had an incontinence score >/= 15. All pre-operative ultrasound images were hypoechoic which correlated with the surgeon's intraoperative findings of scar. The operative appearance included two Type 1, four Type 2, and eight Type 3 images. Larger pre-operative ultrasound image defects were statistically significantly related to intraoperative Type 3 ultrasound images. At a mean follow up of 7.5 (range 2-16) months the mean postoperative incontinence score was 4.5 (range 0-12). In patients with Type 1 and Type 2 images, the mean postoperative score was 8.6 (range 4-12) whereas in patients with Type 3 it was 1.3 (range 0-5) (P < 0.003); 7 of the 8 patients in Type 3 (87.5%) had an incontinence score 10 mm or villous component). The rate of advanced adenomas in males was 4.9% and in females 0.9% (P=0.0006). Three patients (0.3%) were found to have colorectal cancer. Of 43 diabetic patients, 7 (16.3%) had adenomas whereas 6.5% of the non-diabetic persons had adenomas (P=0.03). The proximal colon was investigated in 34 patients by DCBE and/or colonoscopy and the rate of proximal adenomas in patients with neoplastic findings in screening sigmoidoscopy was 5/34 (14.7%). Concerning the experience of screening sigmoidoscopy, 56 persons (93%) found bowel preparation easy and 4 unpleasant, 55 (92%) experienced either no or mild discomfort during the endoscopy whereas 5 found the examination painful; 59 people (98%) said that they would participate again in screening sigmoidoscopy.CONCLUSIONS: In this study of screening sigmoidosopy, the first published in Finland, adenoma rate was 6.9%, advanced adenoma rate was 2.8% and cancer rate was 0.3%. Males had more hyperplastic polyps and advanced adenomas than females. Diabetic people had more adenomas than non-diabetics. Experiences of the people screened were positive and nearly all said that they would participate again in screening sigmoidoscopy. PMID- 12780614 TI - Diverticular disease of the colon and concomitant abnormalities in patients undergoing endoscopic evaluation of the large bowel. AB - OBJECTIVE: To study the prevalence of diverticula and presence of concomitant pathology in consecutive patients undergoing endoscopic examination of the colon. METHODS: A cross-sectional analysis of the endoscopy reports of all patients sent for endoscopic evaluation of the colon in a period of 8.5 years. RESULTS: A total of 9086 endoscopies were performed. Of these 2259 (24.7%) were undertaken for follow-up. Diverticula were seen in 1849 patients (27%) (739 male, 1110 female, mean age 69 year). In 4978 patients (73%)(2162 male, 2816 female, mean age 52 year) no diverticula were seen, of these 2303 (46%) had no abnormalities in their colon. Patients with diverticula were significantly older, 69 vs 46.7 years (P < 0.001). No difference was present in gender. Colorectal cancer and inflammatory bowel disease were significantly more common in patients without diverticula, while polyps were more often seen in patient with diverticula. Patients with diverticula had a significantly lower incidence of colorectal cancer. In addition, the number of cancers located proximal to the splenic flexure was statistically higher in the group of patients presenting with diverticula (P < 0.001). CONCLUSIONS: The overall prevalence of diverticular disease in patients undergoing endoscopy is 27%, and increases with age. Patients with diverticulosis have significantly lower incidence of colorectal cancer and if cancer is detected then it is more common proximal to the splenic flexure. PMID- 12780615 TI - Colorectal cancer in patients with chronic renal failure: the effect of dialysis or renal transplantation. AB - OBJECTIVE: This study reports the characteristics and surgical outcome of colorectal cancer in patients with chronic renal failure treated either by renal transplantation or dialysis. METHODS: Two thousand four hundred and seventeen patients with CRF (1387 transplant and 1030 dialysis) were treated at The Queen Elizabeth Hospital, South Australia, between January 1967 and September 2000. RESULTS: Fourteen have developed colorectal cancer and had their clinical records reviewed. Nine patients were renal transplant recipients and 5 patients were treated with dialysis. Eight patients had surgery. Chemotherapy was given palliatively to 3 patients. One patient died in hospital postoperatively. Seven patients with a renal transplant had notes available. Six presented with late stage disease and all were dead within 9 months of presentation from their disease. In the dialysis patients, tumour stage at presentation was not so advanced and although all the patients have died, the cause of death was secondary to the colorectal cancer in only 2 patients. CONCLUSIONS: This study highlights the poor outcome of patients who have had renal transplant and develop colorectal cancer. Immunosuppression and late diagnosis are implicated in this poor outcome. PMID- 12780616 TI - An analysis of the accuracy of P-POSSUM scoring for mortality risk assessment after surgery for colorectal cancer. AB - OBJECTIVE: To compare predicted vs. observed mortality using P-POSSUM and determine whether this scoring system reflected observed surgical outcomes. PATIENTS AND METHODS: Consecutive patients who underwent a colorectal resection for cancer over a 12-month period were scored using P-POSSUM to estimate mortality risk and POSSUM to calculate predicted morbidity. This was then compared with observed outcomes. The main outcome measurements were 30 day postoperative mortality and postoperative morbidity. RESULTS: 173 patients underwent a colorectal resection for cancer during the study period. The observed mortality rate was 8.7% compared with a P-POSSUM predicted mortality rate of 15.6% (P < 0.01). Observed morbidity was 29% whereas the predicted morbidity rate was 32% (P=0.1). CONCLUSIONS: P-POSSUM allows for standardization of physiological and operative variables. Risk of postoperative mortality may be overestimated by the scoring system however. PMID- 12780617 TI - Tailgut cyst associated with a pilonidal sinus: an unusual case and a review. AB - Tailgut cysts are rare congenital abnormalities in the retrocaecal/presacral region. Clinical diagnosis is difficult and delayed and they can present in childhood and adult life with a variety of clinical symptoms and complications. Differential diagnoses include, rectal duplication cysts, cystic teratoma, epidermal cyst, anal gland cyst and anal gland carcinoma. Magnetic resonance imaging has recently become the modality of choice to image these cysts. Although Tailgut cysts rarely undergo malignant transformation, early surgical resection is presently considered the treatment of choice. Here we report the case of a 34 year old gentleman with a Tailgut cyst associated with a Pilonidal sinus, and review of the literature. We believe that the high incidence of complications associated with operations in the presacral region should be weighed against the generally benign course of these lesions, especially with the quality of modern imaging technology. PMID- 12780618 TI - Inhibition of proliferation of a colon cancer cell line by indole-3-carbinol. AB - OBJECTIVE: To observe the effect of Indole-3-carbinol (I3C), a naturally occurring component of cruciferous vegetables, on cell proliferation of a colon cancer cell line. METHODS: Cell proliferation was measured using three different methods; 3H-thymidine incorporation, cell count and colourimetric assay. RESULTS: Each method of measurement revealed that I3C significantly reduced cell proliferation at concentrations of > 0.1 mM. CONCLUSION: The present study demonstrates for the first time the capacity of indole-3-carbinol to inhibit cell proliferation of a colon cancer cell line. PMID- 12780619 TI - Diverticular colitis - therapeutic and aetiological considerations. AB - OBJECTIVE: Treatment of diverticulitis may change when associated with mucosal inflammation of either ulcerative-like or Crohns-like colitis. To determine effective treatment of diverticular colitis, four cases were analysed. Mechanisms to account for the colitis associated with diverticulitis are put forward. PATIENTS AND TREATMENT: Four cases had colitis and active diverticulitis established by clinical, colonoscopic or CT evidence of active inflammation. Biopsies confirmed mucosal inflammation: two with ulcerative colitis-like and two with granulomata suggestive of Crohns-like colitis. Treatment for colitis in all cases included sulphasalazine and steroids in two cases. RESULTS: Colitis subsided in three cases but one had continuing inflammatory polyps and one case did not resolve. Segmental resection was performed in two cases one with continuing colitis and one with inflammatory polyps. No further attacks of colitis have occurred since the initial observations were made or operations performed. CONCLUSION: An association of colitis with diverticulitis has been confirmed by present observations and case reports of others. Colitis requires medical treatment and if inflammation fails to resolve, segmental resection is indicated. Diverticular colitis, either ulcerative colitis-like or Crohns-like, is part of the spectrum of acute diverticulitis. PMID- 12780620 TI - Hyperplastic polyposis of the colon and rectum. AB - The significance of multiple hyperplastic polyps in relation to the risk of colon cancer is unknown although recent investigation suggests a causative link. We have prospectively identified a small but distinct group of patients that also suggests an association. These patients have either numerous (usually more than twenty, in sites other than the rectosigmoid alone) or large (greater than 1 cm) hyperplastic polyps, in association with either adenomatous polyps, polyps of mixed pathology or carcinoma of the colon and rectum. Additionally, there is frequently a first or second degree family history of colorectal carcinoma. PMID- 12780621 TI - Sigmoid cancer adherent to the uterus during pregnancy: case report. PMID- 12780622 TI - Correction of symptomatic ano-rectal varices with circumferential stapled anoplasty. PMID- 12780623 TI - Colorectal subspecialization in a DGH. The way forward! PMID- 12780626 TI - Trials & tribulations. PMID- 12780627 TI - Early cellular events in colorectal carcinogenesis. AB - Colorectal cancer develops through a multistage process recognizable at a histopathological level by progression from normal mucosa to invasive carcinoma (the adenoma-carcinoma sequence). For many years, it has been hypothesized that increased cell proliferation in the colonic crypt represents the earliest recognizable stage in this sequence. This perspective is now changing. While several human studies have reported increased crypt cell proliferation in samples from at-risk patients, there are many inconsistencies and paradoxes in their conclusions. In addition, it is appreciated that the process of apoptosis (programmed cell death) is vital for normal crypt homeostasis and its impairment may be an early event in the neoplastic process. It is now believed that aberrant crypt foci (ACFs) represent the earliest step in colorectal carcinogenesis. Two ACF types are identifiable: hypercellular and dysplastic. Increased proliferative activity may be seen in both, but the dysplastic entity is most relevant to carcinogenesis. Animal and human studies support the notion that ACFs grow by crypt fission leading to the formation of microadenomas. Adenomas are monoclonal expansions of an altered cell, but very early lesions may be polyclonal. There are outward and inward theories of polypoid growth, and evidence to support both mechanisms. The ACF assay has become a useful tool to detect carcinogens in animal studies but has been less frequently used in human studies. For future cancer chemopreventive and risk assessment studies in humans, the identification and quantification of ACFs should be considered a more effective intermediate marker of risk than the determination of crypt cell proliferation alone. PMID- 12780628 TI - A review of recent randomized trials in colorectal disease. PMID- 12780629 TI - Intestinal spirochetosis: clinicopathological features with review of the literature. AB - OBJECTIVE: The purpose of this study is to evaluate the clinical presentation of patients with intestinal spirochetosis, as it is has not been well described in the literature. METHODS: We studied 15 patients with colonic biopsies that showed spirochetosis. The study group consisted of 11 males and 4 females, 10 of the males were heterosexual and one was homosexual. None of these patients were clinically immunocompromized. A colonoscopy was performed for evaluation of different symptoms. The histological diagnosis of spirochetosis was made on H&E examination and confirmed by Giemsa and/or Steiner stain. There was no evidence of associated active or any specific colitis in any of the cases. The control group consisted of 30 patients, 16 males and 14 females that were matched for age and clinical indications for obtaining a colonic biopsy. RESULTS: Colonic mucosa with prominent brush border-like surface colonized with large amounts of spirochetes was noted in all 15 cases in the study group and in none of 30 cases in the control group. The clinical presentation in patients with spirochetosis was compared to the control group. Of the spirochetosis patients 86% presented with some form of clinical symptoms compared with 13% of controls. These symptoms included chronic watery diarrhoea in 40% spirochetosis patients vs 5% in controls; a change in bowel habit was present in 33% of spirochetosis patients vs 3.3% in controls. The endoscopic appearance in spirochetosis patients was reported as normal in six patients, 'polyoid' in seven patients, erythematous in one patient and 'lesion' in one patient. CONCLUSION: These findings suggest that colonic spirochetosis affects a heterogenous group of patients, the majority of whom presented with gastrointestinal symptoms. The variable clinical findings may be related to the infecting organism and the condition may not be harmless in all patients. PMID- 12780630 TI - Colonic malignancy arising in colitis - a single unit experience. AB - OBJECTIVES: Colorectal malignancy complicating inflammatory bowel disease constitutes 1% of all colorectal malignancies. Although its overall numbers are low it represents the greatest cause of colitis related mortality in these patients. This paper describes the management of 24 patients presenting to a single unit over a period of 10 years. METHODS: The names of patients were collected prospectively when they presented with malignancy. Clinical details were collected by retrospective review of charts. RESULTS: In all, 24 patients with 27 malignancies were identified. The median age of presentation with malignancy was 56 years. Most patients were treated with proctocolectomy. Other patients were treated with segmental colectomy. In these patients the surgical procedure was dictated by the stage of the cancer, the age and comorbid state of the patient and the severity of ongoing colitis. CONCLUSIONS: Malignancy arising in colitis will constitute only a small part of a colorectal practice. The optimum method for detecting early, and potentially curable, disease has not been defined. Surgery should be tailored to the individual needs of the patient. PMID- 12780631 TI - Outcome of argon beam and LASER ablation of large rectal adenomas. AB - OBJECTIVE: Endoscopic ablation of large rectal adenomas is being increasingly used as primary treatment. Despite the avoidance of general anaesthesia and the prevention of more major procedures, patients undergoing endoscopic ablation have the disadvantage of multiple treatment sessions and the lack of adequate tissue sample for complete histological study. The aim of this study was to analyse the outcome of all patients with large rectal polyps treated with endoscopic ablation. PATIENTS AND METHODS: Between 1993 and 1998, 29 patients who underwent endoscopic ablation of large rectal adenoma were identified. All their case notes were analysed and information was collected on recurrence, treatment episodes, complications, the incidence of carcinoma and the necessity for further procedures. RESULTS: At a median 40 (range 4-67) months follow-up, 41% of patients had recurrence of their adenoma and 14% had been diagnosed with adenocarcinoma. Only 24% of patients had been discharged while 21% were clear but were still under surveillance. Seven (24%) patients had complications, 6 stenosis and one severe bleeding. All stenosis occurred in patients who had more than 10 treatment sessions. In all, 31% of patients needed further endoanal or abdominal surgery and the median time to making this decision was 28 (range 4-66) months. There were no deaths. CONCLUSION: Laser and argon ablation of large rectal adenomas has proved very disappointing. It should be reserved for patients who are unfit to undergo general anaesthesia. PMID- 12780632 TI - Preliminary evaluation of United Kingdom National Referral Guidelines for lower gastrointestinal tract cancer. AB - OBJECTIVE: To evaluate the efficiency of the criteria proposed by the National Referral Guidelines at selecting patients with lower gastrointestinal tract cancer for urgent outpatient assessment when compared with the standard method for prioritization. PATIENTS AND METHODS: Consecutive patients aged 50 and over referred to a colorectal unit were offered urgent or non-urgent outpatient appointments on the basis of the contents of the referral letter. Clinical information obtained during the outpatient consultation was used to identify those patients that satisfied the guideline criteria for urgent referral. Patients were investigated and the findings recorded. RESULTS: Of 247 patients who completed their investigations 18 were found to have lower gastrointestinal tract cancer. Urgent outpatient appointments were offered to 115 patients of whom 14 had cancer (P = 0.0067, sensitivity=78%). A total of 119 patients satisfied the guideline criteria for urgent referral, including 17 of the patients who were found to have cancer (P < 0.0001, sensitivity=94%). CONCLUSION: The criteria used in the guidelines provide an effective method for selecting patients with cancer for urgent assessment. Application of the guidelines by general practitioners can be recommended. PMID- 12780633 TI - The effectiveness of new criteria for colorectal fast track clinics. AB - INTRODUCTION: Fast Track Colorectal Clinics are becoming a requirement to meet the government's 10-day rule for patients with suspected cancers to be seen by a specialist. However, such clinics soon tend to get overwhelmed by huge numbers of referrals, many of them inappropriate. The Association of Coloproctology of Great Britain and Ireland has published criteria for appropriate referral. We evaluated the sensitivity of these criteria and attempted to recommend appropriate changes. METHODS: Data of 50 consecutive colorectal cancers from our DGH since January 2000 has been collected in relation to demographics, presenting signs and symptoms, haemoglobin levels as well as treatment. The new Criteria were then applied strictly to these cases and we evaluated whether these patients would have been eligible for fast track referral, if these criteria had been in force since January 2000. RESULTS: Forty-one (82%) of 50 patients would have been eligible for referral as follows: 11 (22%) rectal bleeding with diarrhoea; 8(16%) persistent diarrhoea without bleeding (>60 years), 12 (24%) bleeding without anal symptoms (>60 years), 0 (0%) palpable right sided abdominal mass, 7 (14%) palpable rectal mass, 16 (32%) iron deficiency anaemia (<11 g/dl in men & <10 g/dl in women), 10 (20%) patients qualifying on more than one criterion. However, 9 (18%) patients had presenting features that would have excluded them from a fast track referral as follows: 4 (8%) only abdominal pain + weight loss (48, 54, 72, 75 years old), 2 (4%) change in bowel habit + no rectal bleeding (54, 57 years old), 1 (2%) palpable mass at a site other than the right side of the abdomen. CONCLUSIONS: We feel that this study, although small, highlights the dangers of having very rigid criteria for such clinics. The new criteria though high in specificity have low sensitivity for safety. We recommend some modifications to the criteria as follows: rectal bleeding without anal symptoms >50 years instead of 60 years; patients with a palpable mass anywhere on the abdomen; and the adddition of a new criterion of abdominal pain with weight loss. These modifications would significantly increase the sensitivity from 82% to 94%. PMID- 12780634 TI - The role of anal ultrasound in the management of anal fistulas. AB - OBJECTIVE: To assess the accuracy of anal ultrasound (AUS) for anal fistulas, and the impact of routine pre-operative AUS on their surgical management. METHODS: Pre-operative AUS was performed in 38 consecutive patients with an anal fistula using a 10-MHz Bruel & Kjaer probe. All patients underwent subsequent examination under anaesthetic (EUA) with documentation of the anatomy of the fistula before the surgeon was shown the AUS results. Agreement between AUS and EUA findings and any modification to the surgical treatment was recorded. RESULTS: There was 84% agreement between AUS and EUA findings regarding presence and site of fistulas. One fistula not seen at AUS was found at EUA, and 5 fistulas seen on AUS were not demonstrated at EUA. AUS influenced the surgery undertaken in 9/24 (38%) patients; demonstrating occult sphincter defects (2 patients), reclassifying fistulas from low to higher fistulas (3 patients), deciding a surgical treatment open to doubt (2 patients) and helping identify an obscure fistula not initially found at EUA (2 patients). CONCLUSIONS: Accuracy of AUS in the assessment of anal fistulas is confirmed. Operative management is influenced in 38% of cases, usually towards more conservative treatment. We recommend the use of pre operative AUS in the assessment of anal fistulas. PMID- 12780635 TI - Flexible sigmoidoscopy in general practice. AB - OBJECTIVE: Flexible sigmoidoscopy (FS) is increasingly being accepted as the method of choice for initial investigation of rectal bleeding and other lower gastrointestinal symptoms. The aim of this study was to assess the feasibility of FS in general practice and to compare the yield, cost and efficiency of a service provided by a consultant surgeon and a General Practitioner (GP). SUBJECTS AND METHODS: A prospective study of FS was undertaken on 430 unsedated patients with symptoms suggestive of lower bowel disease in two general practices. RESULTS: The yield was comparable to hospital based data. Fifteen (3.5%) cancers were detected, of which 40% were Dukes A, and 46 (10.7%) adenomatous polyps. Cost per patient examination by a GP including capital costs, depreciation of equipment, and training was calculated to be cheaper or at least equivalent to a hospital based service but inefficient due to the low numbers examined per month. There were no complications and no missed cancers after a minimum follow up of 2 years. CONCLUSIONS: The results suggest that FS can be safely performed by a suitably trained GP in health centres, but better utilization of equipment and value for money would be obtained by providing the service for a group of practices rather than a single practice as in this study. PMID- 12780636 TI - Minimally invasive surgery for ileo-colic Crohn's disease. AB - OBJECTIVE: Patients requiring surgery for Crohn's disease are likely to undergo further surgery due to recurrent disease. A prospective study of laparoscopic assisted surgery for ileo-colic Crohn's disease is reported. METHOD: Forty-one patients (26 female; median age 35 years) with ileo-colic Crohn's disease, without evidence of either fistula or abscess formation underwent laparoscopic assisted surgery. All had medical therapy, including steroids, but had failed to respond or relapsed. Sixteen patients (39%) had previous surgery for Crohn's disease and 26 (63.4%) had previous abdominal surgery. Laparoscopic procedures (n=42) included ileocaecal/ileocolic resection (n=39), small bowel resection (n=1) and strictureplasty (n=2). RESULTS: The median operating time was 90 min (range 60-180). There were 6 conversions (14.6%). Of the 35 patients whose operation was completed laparoscopically, the median postoperative stay was 5 days (3-9) and the median time to full activity was 20 days (7-49). No major complications or death were recorded. The median follow up (34 out of 35) after surgery is 15.3 months (1-55). Twenty-four patients remain symptom free and 4 have minimal symptoms with no clinical or radiological evidence of recurrence of Crohn's disease. Six have however, developed recurrence requiring medical (n=5) and surgical (n=1) treatment. CONCLUSION: Laparoscopic-assisted surgery for Crohn's disease is feasible. It has the advantage of minimizing hospital stay and promoting early recovery in a group of patients who are likely to have further surgery due to the nature of the disease. PMID- 12780637 TI - Preperitoneal approach to parastomal hernia with coexistent large incisional hernia. AB - OBJECTIVE: To assess the outcome of preperitoneal mesh repair of complex incisional herniae incorporating a stoma and large parastomal hernia. METHODS: From 1994 to 1998, symptomatic patients who had repair of combined incisional hernia and parastomal hernia were reviewed. Body mass index, co-morbidity, length of hospital stay, patient satisfaction and outcomes were recorded. RESULTS: Ten patients (seven females and three males), mean age 62 (range 48-80) years underwent primary repair. All had significant comorbidities (ASA grade 3) and mean body mass index was 31.1 (range 20-49). Median hospital stay was 15 (range 8 150) days. Complications were of varying clinical significance (seroma, superficial infection, major respiratory tract infection and stomal necrosis). There were no recurrences after a mean follow up of 54 (range 22-69) months. CONCLUSION: The combination of a parastomal hernia and generalised wound dehiscence is an uncommon but difficult problem. The application of the principles of low-tension mesh repair can provide a satisfactory outcome and low recurrence rate. This must be tempered by recognition of the potential for significant major postoperative complication. PMID- 12780638 TI - Patient's perceptions of biofeedback therapy for obstructive defaecation. AB - OBJECTIVE: The aim of this study is to assess the long-term outcomes of a biofeedback treatment programme for obstructive defaecation up to five years after completion of treatment, reporting from the patients' perspective the subsequent progression of defaecation difficulties and the perceived usefulness of the biofeedback training and its components. METHODS: A biofeedback program for obstructive defaecation at a major hospital in Sydney provided 37 patients with reading material, taught defaecation techniques, and prescribed simulated defaecation and anal relaxation exercises to practice between their monthly biofeedback sessions. Twenty-nine of these patients were contacted by telephone an average of 23 months after last attending and asked to rate the present severity of their condition, changes since last attending biofeedback, and which aspects of the program they found most helpful. RESULTS: Of patients who completed the program, 70% achieved at least a 15% improvement in perceived symptom severity. Those who completed the program and achieved benefit were more likely to name the exercises as the most helpful program component. At the time of follow-up, however, there was no difference in perceived symptom severity between program completers and noncompleters. Program completers, and those followed up more than two years after last attending biofeedback, were more likely to report a worsening in symptom severity subsequent to their final treatment session. CONCLUSIONS: This study confirms the favourable short-term outcomes achieved by some patients completing biofeedback treatment for obstructive defaecation. The findings suggest that biofeedback equipment may be economically used in an educative and corrective role, and that other factors such as education, exercises and counselling alone play an important role. Finally, the benefits of biofeedback may not be maintained through long-term follow-up, raising questions as to the natural history of the disorder and the long-term effectiveness of biofeedback treatment. PMID- 12780639 TI - Traumatic 'degloving' of the colon in blunt abdominal injury. AB - We describe two cases of traumatic 'degloving' of the colon in blunt abdominal trauma. This is an extremely rare injury with the potential to present late. The mechanism of injury appears to be a combination of focal blunt abdominal trauma associated with a shearing force. The potential diagnostic dilemma posed by colonic 'degloving' is outlined and following review of the literature we conclude that CT scanning is the most reliable way of detecting such injuries, if emergency laparotomy is not indicated. PMID- 12780641 TI - Short Papers Meeting, Royal Society of Medicine, London, Section of Coloproctology, 28 November 2001. PMID- 12780640 TI - Intra-dermal methylene blue, hydrocortisone and lignocaine for chronic, intractable pruritus ani. AB - OBJECTIVE: To assess the use of intradermal methylene blue, lignocaine and hydrocortisone in cases of chronic pruritus ani refractory to standard primary, dermatological and colorectal care. METHODS: Five ml 1% methylene blue, 100 mg hydrocortisone and 15 ml 1% lignocaine were injected into the peri-anal skin of 25 patients with chronic pruritus ani which had proved refractory to standard care. Clinical and telephone follow-up was undertaken. RESULTS: After one injection of the above solution, 16 (64%) of patients were rendered symptom free. Repeat injection in those initial nonresponders ultimately rendered 22 (88%) symptom free overall. Morbidity was 4%. Treatment failure occurred in three patients (12%). CONCLUSIONS: Methylene blue used in solution with hydrocortisone and lignocaine can achieve effective control of pruritus ani in 88% of patients who have failed to respond to standard dermatological, hygiene and surgical treatments. PMID- 12780642 TI - Colitis indeterminate. PMID- 12780643 TI - European news. PMID- 12780646 TI - Editorial. PMID- 12780647 TI - Radiation damage to the rectum and anus: pathophysiology, clinical features and surgical implications. AB - Radiation kills cancer cells by inducing various degrees of deoxyribonucleic acid fragmentation and disruption of intracellular membranes that lead to either immediate or delayed cell death. Although radiation can be effective in destroying cancer, its usefulness is limited by damage to normal tissues that surround the target tumour or those in the path of the radiation beam. The rectum and anus are damaged frequently during radiotherapy for abdominopelvic malignancy, including preresection therapy for rectal cancer. Such damage is often associated with lesions in the perineal skin, genitourinary tract, colon, and small intestine. Surgical intervention often is required for the most severe forms of these complications. PMID- 12780648 TI - A prospective audit of fistula-in-ano at St. Mark's hospital. AB - OBJECTIVE: To conduct a prospective audit of all patients presenting with anal fistula at St. Mark's Hospital during one calendar year and to compare the presentation and outcome of this cohort with previous reports from this institution. PATIENTS AND METHODS: All patients undergoing examination under anaesthetic (EUA) for anal fistula during 1997 were studied. All fistulae were anatomically classified and operative procedures recorded. During a mean follow up period of 14 months details of healing, recurrence and function were gathered. RESULTS: 98 patients with a mean age of 43.7 years were assessed. 86 (88%) patients had fistulae of cryptoglandular (idiopathic) origin. Fistulae were superficial in 11 (11%) patients, intersphincteric in 30 (31%) patients, trans sphincteric in 52 (53%) patients, suprasphincteric in 3 (3%) patients and extra sphincteric in 2 (2%) patients. 49 (50%) fistulae were classified as complex. Eradication of fistulae with preservation of function was achieved in 89 (91%) patients. Fistula recurrence occurred in 4 (4%) cases. Ten (10%) patients had some degree of incontinence, 9 (9%) of whom had undergone previous fistula surgery. Nine (9%) patients still had setons in situ at the end of the follow-up period. CONCLUSIONS: A greater proportion of difficult fistulae was seen during the year compared with previous studies from St. Mark's. Despite this a satisfactory outcome was achieved in the vast majority with a relatively low rate of disturbed function. Previous fistula surgery is a risk factor for incontinence, which can usually be managed conservatively. PMID- 12780649 TI - Successful treatment of chronic anal fissure with diltiazem gel. AB - OBJECTIVE: Diltiazem is a calcium channel blocker that has shown potential to heal chronic anal fissures. We have prospectively evaluated the treatment of patients with anal fissure with diltiazem gel (SLA Pharma, UK). PATIENTS AND METHODS: All patients with chronic anal fissure from September 1999 to May 2000 were treated topically with 8 mg 2% diltiazem hydrochloride gel (equivalent to 2 cm of gel supplied by SLA Pharma) three times daily. The 23 patients (12 female), median age 45 (range 22-80) years had a 2-36 month (median 6 months) history of fissure. These were associated with a sentinel tag in 9 (39%) patients. RESULTS: The fissure healed in 11 (48%) of 23 patients, including 6 (75%) of 8 patients who previously failed to heal with GTN ointment. There were no recurrences at 3 months and no adverse effects. Of the 12 patients who did not respond to diltiazem, 4 have undergone sphincterotomy. CONCLUSION: Diltiazem gel has a healing rate similar to that observed with GTN ointment but there were no adverse effects, the recurrence rate was lower, and diltiazem was effective in 75% of patients previously treated unsuccessfully with GTN ointment. PMID- 12780650 TI - Effects of lanreotide on the healing of small bowel anastomoses following obstructive ileus in rats. AB - PURPOSE: Lanreotide, a long-acting somatostatin analogue, inhibits intestinal, bile and pancreatic secretions and decreases intestinal motility. The purpose of this experimental study was to evaluate the effects of lanreotide on the healing of intestinal anastomoses following small bowel obstruction. METHODS: Two groups of 16 Wistar rats (average weight 310 g) were used. Basal diameters of ileus were measured prior to the ligation of the bowel, 15 cm from the ileocecal valve. Luminal fluid was also withdrawn proximal to the obstructed bowel for sodium and potassium analysis. Lanreotide was administered intramuscularly in a single dose (5.4 mg/kg) in the first group, while the same volume of saline was used in the control group. 48 h later rats were re-operated upon. Diameters of the obstructed segments were measured, and luminal fluid of the obstructed bowel was withdrawn and sodium and potassium levels were measured. A segment of 1 cm of the obstructed bowel was resected and end-to-end intestinal anastomosis was performed. Rats were sacrificed on day 7 following the second operation. Anastomoses were examined macroscopically and resected including a 2.5 cm of small bowel on either side. Bursting pressures were measured and the specimens were send for histological examination. RESULTS: The diameter of obstructed bowel increased significantly in both groups. The increase was more prominent in the control group (P < 0.001). Total luminal electrolyte contents for sodium and potassium were stastistically higher in the control group compared to the lanreotide group (P < 0.001). Adhesion formation was more extensive in the control group. Bursting pressures were significantly higher in the lanreotide group compared to the control group (P=0.003). Histological examination of anastomoses showed a more profound inflammatory reaction in the control group compared to the lanreotide group while microscopical healing of the anastomoses was almost the same in both groups. CONCLUSIONS: Lanreotide administration in rats with small bowel obstruction decreases significantly distension and electrolyte losses and seems to improve strength of small bowel anastomoses. PMID- 12780651 TI - A prospective study to assess the implementation of a fast-track system to meet the two-week target for colorectal cancer in Somerset. AB - OBJECTIVE: This prospective study assesses the introduction of a fast-track referral system for patients with suspected colorectal cancer. PATIENTS AND METHODS: The referral system was initiated in Yeovil District Hospital and Taunton and Somerset Hospital using six screening criteria to select high-risk patients. Data on all high-risk patients from 1 November 1999 to 30 April 2000 was recorded prospectively. Patients with proven colorectal cancer diagnosed between 1 November 1998 and 30 April 1999 have been identified for comparison. RESULTS: There were 158 new cases of colorectal cancer in total (111 elective, 47 emergency). 59 cases of colorectal cancer were diagnosed from 433 fast-track referrals (53% of total elective cases). Median time from referral to diagnosis in the fast-track group was 11 days vs. 32.5 days for nonfast-track elective patients (P < 0.001). Median time from referral to diagnosis for all elective cases was 17 days vs. 38.5 days for patients presenting one year earlier (P < 0.001). 75% of fast-track patients were diagnosed with colorectal cancer within two weeks, compared with 17% of nonfast-track elective patients (P < 0.001). The proportion of emergency admissions was reduced from 40% to 30% (P=0.07) following the introduction of this system. CONCLUSION: Data from the first six months reveal that over half of the elective cases of colorectal cancer were diagnosed within the fast-track system. The median time from referral to diagnosis in these patients was within two weeks and there has been a significant reduction in the time to diagnosis associated with the introduction of this service. PMID- 12780652 TI - Factors affecting survival after palliative resection of colorectal carcinoma. AB - OBJECTIVE: To determine the factors affecting survival following palliative large bowel resection for colorectal adenocarcinoma. PATIENTS AND METHOD: From the Colorectal Cancer Database of a single institution patients who had a palliative resection of a colorectal cancer from 1980 to 1993 inclusive were identified. Survival curves were constructed using the Kaplan-Meier method. Criteria studied were sex, age at operation, site of tumour, T, N and M status, tumour differentiation, involvement of tumour margins, tumour fixity and the presence or absence of peritoneal, liver or distant metastases. Multivariate analysis of factors was conducted using Cox proportional hazards analysis. RESULTS: Three hundred and seventy-seven patients (232 men, 145 women, median age 64 years) fitted the above criteria. Operative mortality was 5.6%. Crude 6 month survival rate was 71.1% and median survival 10.5 months. Significant factors affecting survival on univariate analysis were - Age (<75 vs. >75 years) (P=0.019); T status (T1/T2 vs. T3/T4) (P=0.039); nodal status (N0 vs. N1/N2) (P=0.0059); distant metastases (P=0.039) or liver metastases (P=0.0058); tumour differentiation (poor vs. moderate/well differentiated) (P < 0.001); involved tumour margins (P < 0.001). Multivariate analysis found the following factors significant: age (P=0.02), liver metastases (P=0.05), distant metastases (P=0.044), T status (P=0.042), nodal status (P=0.0063), tumour differentiation (P < 0.001) and involvement of tumour margins (P < 0.001). CONCLUSIONS: The data suggest that palliative resection of advanced colorectal carcinoma should be considered carefully in patients with advanced age, where distant metastases are present and in cases when primary tumours can not be completely resected. For the remaining patients, palliative resection may be accomplished with acceptable operative mortality and postoperative survival. PMID- 12780653 TI - The results of total mesorectal excision for rectal carcinoma in a district general hospital before the era of surgical specialization. AB - OBJECTIVE: To evaluate the results of rectal cancer surgery performed by a gastrointestinal surgeon in a district general hospital prior to the introduction of specialization, and to compare these to the targets set by the Royal College of Surgeons for specialist units. METHODS: Data collection in 73 consecutive patients (prospective in 53) undergoing elective excisional surgery (sphincter conserving in 77%) for rectal cancer. While adjuvant radio/chemotherapy and pathological assessment evolved over the study period, a standardized surgical technique (total mesorectal excision) was used in all patients. RESULTS: Observed (and recommended) outcome measures were operative mortality 4.1% (<5), anastomotic leak 6.2% (<8), wound infection 2.7% (< 10), pelvic recurrence after curative resection 9.6% (< 10). Temporary defunctioning stomas were used in 32/48 (66.7%) of patients. Other complications, currently without recommended outcomes, were erectile dysfunction (13%), stoma related (7.1%) urinary retention (4.1%), urinary incontinence (2.7%) and benign anastomotic stricture (2.7%). CONCLUSION: All treatment outcome criteria were met. Trained gastrointestinal surgeons outside the setting of a specialist unit can achieve good results with acceptable complication rates. PMID- 12780654 TI - Using quality of life scores to help determine treatment: is restoring bowel continuity better than an ostomy? AB - OBJECTIVE: In colorectal surgery, evaluation of heath-related quality of life (HRQL) has been relatively minimal when compared to other medical specialties. Would the performance of such HRQL evaluations change our decision-making in patient care? In familial adenomatous polyposis (FAP), procedures that restore bowel continuity (i.e. Ileorectal anastomosis or ileal pouch anal anastomosis) are routinely preferred to ileostomy because of the perceived, but unproven, better HRQL. This study evaluates FAP patients who underwent prophylactic colectomy with either permanent ileostomy or 'restored bowel continuity' reconstruction. The functional outcomes of both groups are reported, and the HRQL assessments are compared. METHODS: All FAP patients who underwent (procto) colectomy resection with reconstruction, either restored bowel continuity (BC) or permanent ileostomy (OST), between 1980 and 1998 were studied. Functional data were obtained by questionnaire and medical record review. HRQL was assessed by 2 validated instruments - the SF-36 Physical and Mental Health Summary Scales and the SF-36 Health Survey - which measure physical summary (PSF) and mental summary functioning (MSF) as well as eight separate health quality dimensions including health perception (HP), physical (PF) and social functioning (SF), physical (PR) and emotional role limitations (ER), mental health (MH), bodily pain (BP), and energy level (E). RESULTS: Results were obtained in 54 patients; bowel continuity (44), ileostomy (10). Mean patient age was 39 years, mean follow up time was 10.5 years. Mean patient age at operation was 28 years. Functional results for BC included number of bowel movements/day (6.7), leakage (30%), having to wear a pad (11%), perianal skin problems (25%), food avoidance (68%), and inability to distinguish gas (27%). Functional results for OST were routinely excellent. Results of the HRQL surveys reveal no significant differences for BC vs OST (HP: 67 +/- 28 vs 79 +/- 39; PF: 91 +/- 14 vs 90 +/- 17; SF: 86 +/- 23 vs 97 +/- 5; PR: 79 +/- 34 vs 83 +/- 40; ER: 86 +/- 28 vs 88 +/- 27; MH: 77 +/- 19 vs 82 +/- 14; BP: 78 +/- 24 vs 71 +/- 32; E 60 +/- 21 vs 58 +/- 18, respectively). CONCLUSION: Although the perceived quality of life for ileostomy patients is generally worse than the 'restored bowel continuity' group, the measured HRQL is the same for both groups. These results suggest that a permanent ileostomy should be included as a viable and appropriate first line treatment option for FAP patients after resection. This study also suggests that HRQL should play a greater role in the evaluation of care and treatment in colorectal surgery. PMID- 12780655 TI - Untreatable faecal incontinence: colostomy or colostomy and proctectomy? AB - PURPOSE: To determine the frequency of rectal symptoms and secondary proctectomy in patients undergoing elective permanent end sigmoid colostomy for faecal incontinence and determine risk factors. METHODS: A retrospective chart review of patients undergoing elective end sigmoid colostomy for faecal incontinence at St Mark's Hospital between January 1991 and December 1998. Patients were divided into three groups: A, symptoms leading to subsequent proctectomy; B, symptomatic but avoiding proctectomy; C, asymptomatic. RESULTS: There were 44 patients (80% women, average age 57 years): Group A 12 (27%); Group B 13 (30%); Group C 19 (43%). Group A were on average younger than Group C (45 years vs 64 years, P < 0.05). No other risk factor for symptoms or proctectomy was evident, and in particular a prior history of obstructed defaecation/anal digitation was not related. Only half the patients undergoing proctectomy had histological evidence of defunctioned proctitis in the resected rectum. CONCLUSIONS: Data are insufficient presently to recommend primary proctectomy in this group of patients (even if it were to be performed laparoscopically). PMID- 12780656 TI - Transanal full-thickness excision of rectal tumours: should the defect be sutured? a randomized controlled trial. AB - PURPOSE: It is generally recommended that the defect, after full thickness total wall excision of a tumour located in the extraperitoneal part of the rectum, should be sutured. There is a lack of controlled studies however, supporting this approach. The aim of this study was to compare the results obtained in patients after peranal local excision of rectal tumours whose defect were sutured with those that were not. METHODS: 44 patients were prospectively randomized to group A: The defect is closed; Group B: Defect left un-sutured. Pre-operative test were digital examination, proctoscopy and endorectal ultrasound. Local full-thickness excision was performed mainly with the Transanal Endoscopic Microsurgery (TEM) equipment, but for cases near the anal verge a Parks' retractor was used. Data recorded were operation time, blood loss, hospital stay and early and late complications. The first postoperative assessment was planned at 1 month and then every three months until 18 months of follow-up. Result for 40 patients (21 from group A; 19 from group B) were analysed. There were no differences between groups regarding age, sex, location of the tumour and specimen's size. RESULTS: The intra-operative loss of blood was 22 ml for group A and 39 ml for B, the difference was not significant. The mean operation time was slighter longer for group A (93 min) than for group B (77 min) but not statistically significant. For both group the mean hospital stay was of 4[2-7] days. No differences in early or late complications could be demonstrated. CONCLUSION: The present study suggests that there is no difference between these two practices in terms of intra operative results and outcome. PMID- 12780657 TI - Altered serotonin immunoreactivities in the left colon of patients with colonic inertia. AB - BACKGROUND: Serotonin is an important positive regulator of colonic motility and transit. Its quantity and distribution in the left colon could be abnormal in patients with colonic inertia (CI) and contribute to the disease. AIM: To evaluate serotonin positive cells and immuno-reactivities in the mucosa, submucosa and muscularis propria of the left colon from patients with CI was compared to a control group. PATIENTS AND METHODS: Nineteen patients who underwent subtotal colecotomy for CI were assessed. The control group consisted of 15 patients who underwent left hemicolectomy for colonic cancer; histologically normal tissue specimens were used. Immunohistochemical staining for serotonin was performed. In the mucosa, the average number of serotonin positive cells per microscopic field (200 x ) was determined. The positively stained area (square pixels) in the mucosa, submucosa and muscularis propria per microscopic field (200 x ) was calculated utilizing a computer image analysis program. RESULTS: In the mucosa, both the number of serotonin positive cells and positively stained area were significantly higher in the patient group than in controls (P < 0.05). The difference in serotonin positive area in the submucosa in the CI group compared to the control group was not statistically significant. There was a very significant correlation between the serotonin positive area in the submucosa and muscularis propria in controls (r=0.65, p < 0.01), but not in patients with CI. CONCLUSION: The increased serotonin level in patients with CI may contribute to the disease or be an adaptive response to some other pathology. The lack of a positive correlation in serotonin levels between the submucosa and muscularis propria in CI patients suggests that the coordinated distribution of serotonin may be disrupted in CI. PMID- 12780658 TI - Objective assessment of morbidity and quality of life after surgery for low rectal cancer. AB - OBJECTIVE: To determine whether an anterior resection for low rectal cancer is associated with less morbidity and a better quality of life than an abdominoperineal resection. PATIENTS AND METHODS: Fifty-three patients who had anterior resection for low rectal cancer were individually matched with 53 other patients, who had an abdominoperineal resection during the same period of time, for gender, age and socioeconomic status. There was no evidence of tumour recurrence in these patients. All patients answered three quality of life questionnaires at least one year after surgery: Short Form 36 version 2, EORTC QLQ-C30 and EORTC QLQ-CR38. Details on short-term and long-term complications were collected prospectively. RESULTS: There was no significant difference in both the short-term and the long-term complication rates between the two groups of patients. There was also no difference in the global quality of life score on the QLQ-C30. However, patients who had an anterior resection had a better perception of body image than those who had an abdominoperineal resection, as measured on the QLQ-CR38 questionnaire (P=0.009), but were more prone to suffer from constipation (P=0.001). CONCLUSIONS: The overall similarities in quality of life between the two groups is probably a reflection of better stoma care, although problems with body image still persist in stoma patients. PMID- 12780659 TI - Outlet obstruction due to anismus and rectal hyposensation: effect of biofeedback training. PMID- 12780660 TI - Sulindac and tamoxifen in the treatment of desmoid tumours in patients with familial adenomatous polyposis. PMID- 12780663 TI - Chronic rejection with or without transplant vasculopathy. AB - BACKGROUND: Chronic allograft nephropathy (CAN) is defined and graded in the Banff '97 scheme by the severity of interstitial fibrosis and tubular atrophy. It has been denoted that chronic rejection can be diagnosed if the typical vascular lesions are seen, consisting of fibrointimal thickening. We observed several patients who developed CAN without vascular changes or signs of cyclosporine toxicity. Therefore, we assessed the risk factor profiles of CAN with and without transplant vasculopathy. METHODS: A cohort of 654 cadaveric renal transplants performed between 1983 and 1997 that functioned for more than 6 months was studied. Fifty-four transplants had CAN defined by a significant decline in renal function together with interstitial fibrosis and tubular atrophy without signs of cyclosporine nephrotoxicity or recurrent disease. Using the Banff chronic vascular (CV) score, 23 of 54 cases (43%) had a chronic vasculopathy score of 0 or 1 whereas 31 cases (57%) had a CV score of 2 or 3. Applying multivariate logistic regression, predictor variables of the two groups were compared with 231 transplants with a stable function for at least 5 yr. RESULTS: Graft histology was obtained at a mean of 2.4 and 2.9 yr after transplantation in the group with or without vasculopathy, respectively. Acute rejection episodes (AREs) after 3 months post-transplantation were the strongest risk factor for both forms of CAN, odds ratio (OR) 14.7 (6.0-36.0). CAN with vasculopathy was also associated with transplants performed in the 1980s, OR 4.95 (1.65-14.9) and with creatinine clearance at 6 months, OR 0.58 (0.44-0.75) per 10 mL/min increase. In contrast, young recipient age, OR 0.69 (0.47-0.99) per 10-yr increase, and the presence of panel reactive antibodies at the time of transplantation, OR 1.26 (1.08-1.47) per 10% increase, were independent risk factors for CAN without vasculopathy. CONCLUSIONS: After exclusion of cyclosporine toxicity or recurrent disease CAN occurred without moderate or severe transplant vasculopathy in 43% of the cases. The correlation with young recipient age, sensitization and late ARE suggest an immune pathogenesis, consistent with chronic rejection. PMID- 12780664 TI - Influence of SDZ RAD vs. MMF on gastric emptying in renal transplant recipients. AB - SDZ RAD and mycophenolate mofetil (MMF) are increasingly used in the prevention of renal allograft rejection. SDZ RAD, having a macrolide structure, and MMF, known with gastrointestinal side-effects, may have gastric motility modifying properties. Gastric emptying was examined 1 yr after renal transplantation in eight patients taking corticosteroids (CS), cyclosporin A (CsA) and SDZ RAD and six patients treated with CS, CsA and MMF. Comparing the two groups, no significant differences in gastric emptying of solids and liquids were noted. Compared with normal volunteers, solid gastric emptying was faster in the SDZ RAD group and similar in the MMF group. It is concluded that in stable renal transplant recipients treated with MMF, gastric emptying was normal. Because of the impact on drug absorption and gastrointestinal symptoms, further studies are indicated to corroborate the potential prokinetic properties of SDZ RAD. PMID- 12780665 TI - Steroid-free immunosuppression during and after liver transplantation--a 3-yr follow-up report. AB - BACKGROUND/AIMS: Steroids are traditionally used in liver transplantation as a part of a triple or quadruple immunosuppressive regimen. Steroids act non specifically and cause multiple side-effects. Most liver transplantation centers reduce the dosage of steroids and eventually withdraw them after various time intervals. A few steroid-free trials have been recently conducted after liver transplantation but long-term data are not yet available. In addition, in these trials steroids were usually given during surgery. We report the long-term (median = 40 months) follow-up data of a prospective pilot study designed to determine whether liver transplantation could be performed with no steroids at all (neither during nor after surgery). METHODS: Twenty-one consecutive liver transplantations in 20 adult patients between August 1998 and July 1999 were prospectively included in an ab initio steroid-free immunosuppressive protocol. Mean age was 54 yr (40-67 yr). Tacrolimus (through levels, 8-10 ng/mL) and azathioprine (1-2 mg/kg) were started after liver transplantation. Patients were not given steroids during or after liver transplantation except in the event of rejection or in case of tacrolimus or azathioprine toxicity requiring significant dose reduction and/or withdrawal. RESULTS: There has been no case of primary graft dysfunction or non-function. Eleven of 21 liver transplantations (52%) received no steroids throughout the whole study. Rejection developed in five of 21 liver transplantations (23.5%). These rejections responded to standard i.v. steroids (plus ATG in one patient), followed by an oral steroid taper stopped 3 months after rejection. Steroids were transiently given in six liver transplantations for non-immune reasons: two with tacrolimus-induced neurotoxicity, three cases where azathioprine was discontinued, and one for an allergic reaction; four of these six patients are off steroids at last follow-up. The 3-yr graft and patient survival is 95 and 100%, respectively. CONCLUSIONS: Steroids are not necessary in more than 50% of liver transplantations. Steroids were transiently needed to treat acute rejection in 23.5% liver transplantations and for toxicity of calcineurin inhibitors or azathioprine or other reason in 28%. Of the patients who received steroids, the majority (70%) was eventually taken off steroids. This prospective single-center pilot study shows that liver transplantation without steroids is feasible and yields no penalty in terms of acute and chronic rejection, immune graft loss, graft function, patient and graft survival. PMID- 12780666 TI - The first report from the patient outcomes registry for transplant effects on life (PORTEL): differences in side-effects and quality of life by organ type, time since transplant and immunosuppressive regimens. AB - BACKGROUND: Post-transplant patient quality of life (QOL) is affected by a number of different factors. A nationwide patient registry has been established to evaluate QOL and determine the effects of transplant and immunosuppressive regimens on patient outcomes. METHODS: Patients were contacted directly at national meetings, through transplant centers, and patient support groups and invited to participate in the registry. All transplant patients aged 16 and over were eligible to enroll. Patients completed a 100-item self-administered questionnaire consisting of questions about patient demographics, organ functioning, and other post-transplant outcomes. General QOL was measured by the Short form - 12 (SF-12). The Memphis Survey, an instrument developed and psychometrically validated at the University of Tennessee, was administered to patients to evaluate side-effects associated with immunosuppression. Data were analyzed from the first 722 patients who entered the registry. Side-effect profile and QOL outcomes were evaluated by organ type, time since transplant and immunosuppressive regimen. Multiple regression analyses were conducted to determine predictors of post-transplant QOL. RESULTS: When outcomes were analyzed by organ type, there were no differences in SF-12 or total weighted Memphis scores. Analysis by time since transplant demonstrated that side-effects in the mobility domain increased with patient age and time since transplant. Analysis by immunosuppressive regimen focused on cyclosporine and tacrolimus-based regimens congruent with similar classifications reported in previous studies (Pirsch JD et al. Transplantation 1997: 63: 977, Shield CF III et al. Transplantation 1997: 64: 1738). When analyzed by regimen, there were no differences between the groups in terms of patients reporting good to excellent organ function, treatment for rejection, infection, and over-immunosuppression. Statistically significant differences were observed when side-effect profile was analyzed by immunosuppressive regimen. Patients on cyclosporine-based regimens reported greater overall side-effect severity and more problems with mobility and life roles. Cyclosporine patients also reported more problems in the miscellaneous subscale, including high blood pressure, enlarged gums and hair growth, but less trouble with trembling hands. Multiple stepwise regression models identified several side-effect subscales as having profound effects on mental and physical QOL. CONCLUSION: Transplant recipients report good to excellent levels of QOL, however, side-effects associated with immunosuppressive regimens impair post transplant QOL. Problems in certain domains, such as mobility, are found to increase with time since transplant. Tacrolimus-based regimens are associated with fewer and less severe side-effects than cyclosporine-based regimens in key domains that affect post-transplant QOL. PMID- 12780667 TI - Chronic liver disease in kidney recipients with hepatitis C virus infection. AB - BACKGROUND: The prevalence of anti-hepatitis C virus (HCV) positive test is higher among patients in dialysis and in kidney recipients than in general population. Hepatitis C virus infection is the main cause of chronic liver disease in renal transplant patients. Liver biopsy and virological analysis were performed to clarify the grade of liver damage in kidney recipients. METHODS: Renal recipients patients with at least 5 yr under immunosuppression were submitted to clinical and laboratory analysis. Patients who tested anti-HCV positive were candidates to liver biopsy with no regard to transaminase levels. RESULTS: Forty-five patients tested anti-HCV positive and 42 anti-HCV negative. Twenty-six anti-HCV and RNA-HCV positive patients were submitted to liver biopsy. Seventy-three percentage of these patients presented chronic active hepatitis, from these only one patient presented cirrhosis. Only 29% of the anti-HCV positive group presented elevated alanine aminotransferase levels. Anti-HCV positive patients presented longer previous time on dialysis and less rejection episodes than the group anti-HCV negative (p < 0.05). All anti-HCV positive patients but one tested RNA-HCV positive by polymerase chain reaction (PCR). CONCLUSIONS: In this series the prevalence of anti-HCV positive is 51.7%. Most of the patients presented liver damage in histology caused by HCV. However, we found only mild or minimal fibrosis and inflammatory activity grade, despite 10 yr of HCV infection and 5 yr of immunosuppressive treatment. Only one patient presented cirrhosis (4%). Performing serial liver biopsies in a long-term follow-up is needed to clarify the impact of HCV infection in renal transplant patients. PMID- 12780668 TI - The impact of mycophenolate mofetil dosing patterns on clinical outcome after renal transplantation. AB - BACKGROUND: Mycophenolate mofetil (MMF) has proven to be a very effective drug for the prevention of acute rejection following renal transplantation when dosed as prescribed at 2 or 3 g/d. However, circumstances arise in clinical transplantation where the dose must be lowered, either to avoid drug toxicity or because of concurrent infection. The impact on the incidence of acute rejection and graft survival when the MMF dose must be lowered has not previously been investigated. METHODS: In this study, a cohort of 721 kidney transplant recipients who received immunosuppression using MMF in conjunction with cyclosporine and prednisone and OKT3 (n = 425) or Simulect (n = 296) induction were evaluated. Clinical outcomes were compared and contrasted between patients with and without MMF dose changes within the first year post-transplantation. RESULTS: The majority of patients (70.3%, n = 507) had at least one dose change within the first post-transplant year. Compared with the 214 patients who did not have a dose change, these patients had a much higher incidence of acute rejection within the first post-transplant year (23.3% vs. 3.7%, p < 0.001). This resulted in a significantly decreased 3-yr death-censored graft survival (76.3% vs. 88.3%, p = 0.003). The incidence of acute rejection for patients who had a dose change was highest if the dose change occurred within the first post-transplant month (34.4%). The incidence of acute rejection for the dose change patients was influenced by recipient ethnicity (African-American vs. Caucasian) and the type of induction agent used (OKT3 vs. Simulect). CONCLUSION: Altering the dose of MMF within the first post-transplant year correlated with a significantly worse clinical outcome in this cohort of renal transplant recipients. These data suggest that avoidance of MMF dose changes within the first year after renal transplantation would result in improved graft survival. PMID- 12780669 TI - Effect of cyclosporin pharmacokinetics on renal allograft outcome in African Americans. AB - African-Americans (A-As) experience inferior outcome after transplantation compared with other ethnic groups. Bioavailability of cyclosporin (CsA) has been implicated as a possible contributing factor. This paper describes the outcome of 32 A-A recipients of de novo renal allograft who received CsA-based triple immunotherapy according to individual pharmacokinetic profiles. Patients received CsA-microemulsion q 12 h, dosed initially at 3.5 mg/kg (8 am) and 3.0 mg/kg (8 pm). The am and pm doses were independently adjusted to achieve a 12-h area under the concentration-time curve (AUC0-12) of 6600-7200 nghr/mL and morning trough level (C0) of 250-325 ng/mL, respectively. Mean age was 43 +/- 12 yr, 37% (12) female. Mean AUC0-12 in 1 wk, 1, 3, 6, and 12 months were 7810 +/- 1880 nghr/mL, 9057 +/- 2097 nghr/mL, 7674 +/- 1912 nghr/mL, 7132 +/- 2040 nghr/mL, and 6503 +/- 1410 ngl/h with corresponding C0 of 301 +/- 79 ng/mL, 316 +/- 66 ng/mL, 275 +/- 59 ng/mL, 273 +/- 66 ng/mL, and 224 +/- 49 ng/mL, respectively. Acute rejection occurred in two patients (6%) 1 yr after transplantation. Prospective use of CsA pharmocokinetic profiles improves renal allograft outcome in A-As. PMID- 12780670 TI - The effect of calcineurin inhibitors on endothelial function in renal transplant recipients. AB - Endothelial dysfunction is of vital importance, as it may cause ischemia and dysfunction in various organs. Despite, this problem has been well documented in patients with end-stage renal disease (ESRD), there is not enough data considering this issue following renal transplantation. One of the potential causes of endothelial dysfunction in renal transplant recipients may be administration of calcineurin inhibitors. The aim of this study is to evaluate the effects of two different calcineurin inhibitors [cyclosporin A (CsA) and tacrolimus (FK506)] on endothelial function in renal transplant patients. Forty four renal transplant recipients [22 on FK506 (group I) and 22 on CsA (group II)] were studied. Endothelial functions of the brachial artery were evaluated by using high resolution vascular ultrasound. Endothelium-dependent and -independent vasodilations were assessed by establishing reactive hyperemia and using sublingual nitroglycerine (NTG), respectively. Results are presented as percentage change from baseline values. Significant endothelial dysfunction was noted in renal transplant patients treated with CsA. While endothelium-dependent vasodilation was 12.1 +/- 5.1% in group I and it was 6.5 +/- 3.7% in group II (p < 0.001). The increase in brachial artery diameter after sublingual NTG was 20.1 +/- 6.3 and 12.7 +/- 5.6% in groups I and II, respectively. This indicates that the endothelium-dependent and -independent vasodilation of the patients on FK506 is better preserved than the patients on CsA therapy. Besides, blood flow volume (BFV) increase was 51.2 +/- 39.4 and 43.9 +/- 24.3%, in groups I and II, respectively, in reactive hyperemia period (p > 0.05). Post-transplant course of renal transplant recipients is complicated by endothelial dysfunction. This problem is more prominent in patients on CsA therapy, which can predispose these patients to more frequent cardiac complications. PMID- 12780671 TI - The effect of ATG on cytokine and cytotoxic T-lymphocyte gene expression in renal allograft recipients during the early post-transplant period. AB - BACKGROUND: Despite the long history of ATG use, the exact in vivo mechanism of action remains unclear. In the present study, we analyzed the effect of ATG induction therapy on expression of 10 immunologically relevant genes in the early post-transplant period. METHODS: Eight renal allograft recipients received post transplant prophylactic ATG treatment on 10 consecutive days and an additional three patients received treatment on 5, 6, or 7 consecutive days, respectively. Gene expression was measured at the beginning and the end of therapy and normalized to a control gene using Taqman real-time PCR methodology. Results were compared with those of matched control patients. No patients were diagnosed with rejection. RESULTS: ATG-treated patients showed decreases in the expression of cytotoxic T cell genes perforin (-56%, p = 0.03) and granzyme B (-45%, p = 0.01) and cytokine gene IFN-gamma (-75%, p = 0.005), and significant increases in the expression of cytokine genes IL-7 (550%, p = 0.04), IL-10 (275%, p = 0.01), IL-15 (417%, p = 0.03), TNF-alpha (615%, p = 0.01), and TGF-beta (235%, p = 0.02). No significant changes were observed in the control group, with the exception of a decrease in IL-10 expression (-42%, p = 0.01). There were no significant changes in IL-12 or Fas-L expression in either group. CONCLUSION: ATG-induced decreases in the expression of IFN-gamma, perforin, and granzyme B and increases in IL-10 and TGF-beta might be considered beneficial to the recipient, whereas increases in the expression of IL-7, IL-15, and TNF-alpha genes might be involved in immunological processes not effected by ATG that may harm the transplant in the long term. PMID- 12780672 TI - Exercise capacity and muscle structure in kidney recipient and twin donor. AB - Exercise capacity and muscle function are known to be abnormal in patients with renal disease. We present a case study in which we studied a kidney transplant recipient who received a kidney from his identical twin. Testing included maximal exercise testing with measurement of oxygen uptake (VO2max), isokinetic muscle function testing, body composition analysis using dual energy X-ray absorptiometry (DEXA) imaging and quality of life using The Medical Outcomes Short Form (SF-36) questionnaire. Muscle biopsies were also analysed for fibre composition and size and ultrastructure. We found that the twin recipient had lower values for VO2max, muscle strength, muscle mass and quality of life scales (physical domains) compared with the twin donor. Muscle fibre composition was identical in the twins, however, muscle fibres size was reduced in the twin donor in Type I, and Type IIA fibre types. The twin recipient also showed lower fibre density (volume percentage) and greater intrafibrillar space than the donor. The twins were both sedentary, thus we conclude that the low exercise capacity and abnormalities observed in the recipient are characteristics of renal failure, and not the result of physical deconditioning, nor was it related to prednisone therapy, which may negatively affect muscle function. PMID- 12780673 TI - Cyclosporine and its metabolites before and 2 h post-dose: comparative measurements of a monoclonal and a polyclonal immunoassay. AB - The aim of the study was to investigate the better accuracy of the 2-h post-dose (C2) levels of cyclosporine (CyA), compared with the pre-dose (C0) levels and to evaluate the results measured by a monoclonal or a polyclonal immunoassay. The parent compound of CyA in C2 (monoclonal2) was measured in 53 kidney transplant patients by the monoclonal fluorescence polarization method, as well as the parent compound plus metabolites (polyclonal2) by the polyclonal fluorescence polarization method. Also, the parent compound was measured in 21 of the patients for the C0 (monoclonal0), whereas the parent compound plus metabolites in 36, for the C0 (polyclonal0). As level of metabolites was considered the difference between polyclonal and monoclonal values (polyclonal-monoclonal), either in C0 (metabolites0) or in C2 (metabolites2). The ratio polyclonal2/monoclonal2 gave a mean value of 1.7+/-0.2 (mean+/-SD), whereas the mean value of the ratio polyclonal0/monoclonal0 was 2.3+/-0.6, with almost double variation. The mean value of the ratio metabolites2/monoclonal2 was 0.7+/-0.2 and of the ratio metabolites0/monoclonal0 was 1.3+/-0.6. The difference between the two ratios is very significant (p = 0.000001) and they are not correlated with each other (r = 0.18, p = 0.44). The measurements of monoclonal0 and polyclonal0 or monoclonal2 and polyclonal2 are very significantly correlated (r = 0.94, p = 0.000001 and r = 0.97, p = 0.000001, respectively). In C0 the proportion of metabolites is higher than in C2, with a double variation, as the degree of metabolism is diverse. Consecutively, in monoclonal methods, as cross-reactions occur with metabolites, it is more accurate to use the C2 measurement for the evaluation of CyA. The application of both methods, the polyclonal and the monoclonal, could be a useful tool as it gives an estimation of metabolites whose degree of contribution to the immunosuppressive result is difficult to ascertain. Finally, if for reasons of clinical experience, the polyclonal method is used, then the mean therapeutic levels of polyclonal2 are 1.5-1.7 compared with monoclonal2. PMID- 12780674 TI - The use of an anti-CD25 monoclonal antibody and mycophenolate mofetil enables the use of a low-dose tacrolimus and early withdrawal of steroids in renal transplant recipients. AB - BACKGROUND: Reducing calcineurin-inhibitor-induced nephrotoxicity and simultaneously avoiding long-term steroid related side-effects is a desirable goal in renal transplantation. We examined the hypothesis that using anti-CD25 monoclonal antibody induction and mycophenolate mofetil (MMF) would allow the lowering of target pre-dose blood concentrations of tacrolimus immediately after transplantation and subsequently stopping steroids at 5 months. METHODS: Eighty two kidney recipients were enrolled in a single-center study comparing two tacrolimus-based protocols. Group I (n = 41) patients received a standard-dose tacrolimus (blood concentration 10-15 ng/mL) with MMF and a standard dose corticosteroid. Group II (n = 41) patients were treated with a low-dose tacrolimus (blood concentration 5-10 ng/mL) and MMF, a low-dose corticosteroid (stopped after 5 months) and induction with daclizumab. RESULTS: Patient (95.1 versus 100%) and graft survival (92.6 versus 97.5%) at 1 yr were not different between groups. Patients of group II experienced significantly less acute rejections than group I (17.1 versus 41.4% p = 0.03). Delayed graft function occurred less often in group II (5 versus 12% p = 0.43). Graft function at 1 yr was significantly better in group II (serum creatinine 1.49 versus 1.69 mg/dL and creatinine clearance 59.6 versus 49 mL/min; p < 0.05). Corticosteroids could be stopped after 5 months in 82.9% of group II patients. CONCLUSION: A regimen consisting of anti-CD25 monoclonal antibody induction and MMF allows the safe and efficient use of low-target pre-dose trough concentrations of tacrolimus and enables the early discontinuation of steroids. Preliminary results indicate a better 1-yr graft function compared to a normal-dose tacrolimus regimen. PMID- 12780675 TI - Pancreas transplantation in crossmatch-positive recipients. AB - BACKGROUND: Prolonged cold preservation time can unfavorably affect outcome in pancreas transplantation. To reduce this ischemic time, cadaver pancreas grafts, in selected cases, are sometimes transplanted before crossmatch results are known. We report our experience with pancreas transplants in recipients with either current or historically positive T- or B-cell crossmatches. METHODS: Crossmatch-positive pancreas transplants were identified using a computerized database. T-cell crossmatches were performed using an antihuman-globulin augmented complement-dependent cytotoxicity (CDC) test; B-cell crossmatches were performed using an extended incubation CDC test. All patients received anti-T cell induction therapy and either cyclosporine (1987-1993) or tacrolimus-based (1994-2001) immunosuppression. More recent recipients (2000-2001) also received intravenous gamma globulin and postoperative plasmapheresis. RESULTS: Between October 1, 1987 and March 31, 2001, of a total of 1076 pancreas transplants performed, 59 (5.48%) were crossmatch-positive. Of these, 8 had a current T-cell positive crossmatch and 15 had a current B-cell-positive crossmatch. One recipient was both current B- and T-positive, and the rest were past B- and/or T cell positive. One-year pancreas graft survival for current T- and B-cell crossmatch-positive transplants was 63% and 67%, respectively. T- or B-cell crossmatch-negative transplants had a 1-yr survival of 70%. In the T-cell crossmatch-positive group, four grafts are still functioning (follow-up range, 2 12 yr), one patient died with a functioning graft at 4 months, and four grafts failed (one each from pancreatitis, infection, primary nonfunction, and vascular thrombosis). No grafts were lost to rejection. In the B-cell crossmatch-positive group, six grafts are still functioning (follow-up range, 2-11 yr) and nine have failed (four from chronic rejection, three from vascular thromboses, and two from pancreatitis). Crossmatch-positive cases were significantly more likely to be retransplants (70.8%) than crossmatch-negative cases (14.8%, p < 0.0001). In a multivariate analysis, crossmatch positivity did not affect pancreas graft outcome, whereas retransplants had a significant impact on outcome (relative risk 1.84, p < 0.0001). CONCLUSIONS: (: i) Pancreas transplants performed in the setting of a positive current crossmatch may have long-term function. (ii) With current immunosuppressive protocols, graft loss from hyperacute and acute rejection may be prevented in current crossmatch-positive pancreas transplants. Chronic rejection was only seen in B-cell crossmatch-positive cases. (iii) High rates of technical graft loss in crossmatch-positive cases may reflect a high frequency of retransplants in this group. PMID- 12780677 TI - Cost-effective and safe ambulatory long-term immunoprophylaxis with intramuscular instead of intravenous hepatitis B immunoglobulin to prevent reinfection after orthotopic liver transplantation. AB - BACKGROUND: Hepatitis B (HBV)-infected patients receive an anti-HBs immunoprophylaxis [hepatitis B immunoglobulin (HBIG) titre of more than 100 IU/L] in combination with lamivudine to prevent reinfection after orthotopic liver transplantation (OLT). In comparison with intramuscular (i.m.) HBIG, costs for intravenous (i.v.) HBIG are found to be extremely high. We therefore studied patients' outcome (i) after a switch from i.v. to i.m. HBIG and (ii) the outcome after the patients were initially treated with i.m. HBIG after discharge from the hospital. METHODS: (i) Six outpatients were switched from 2000 IU i.v. HBIG (Hepatect) administered every 2 wk to 2000 IU i.m. HBIG (Hepatitis-B Immunoglobulin Behring) given once a month. (ii) Six other outpatients were directly treated with i.m. HBIG every 4 wk after OLT. All patients also received 100 mg lamivudine/d. RESULTS: Patients switched from i.v. to i.m. HBIG had stable anti-HBs titres (i.v. HBIG: 180 +/- 37 IU/L vs. i.m. HBIG: 173 +/- 23 IU/L). Patients directly treated with i.m. HBIG also had sufficient anti-HBs titres (176 +/- 31 IU/L). Intramuscular application of HBIG was well tolerated by all patients and no side-effects were observed in patients receiving i.m. HBIG. In comparison with the protocol using i.v. HBIG, the costs of i.m. treatment were 60% lower. CONCLUSION: Long-term administration of i.m. HBIG saves up to 60% of the usual costs for i.v. prophylaxis of HBV reinfection in patients after OLT. In combination with lamivudine, long-term i.m. HBIG therapy is as efficient as i.v. HBIG treatment, but its lower costs clearly favour its use in preventing HBV reinfection after OLT. PMID- 12780676 TI - Two- to three-fold increase in blood tacrolimus (FK506) levels during diarrhea in liver-transplanted children. AB - BACKGROUND: The diagnosis and treatment of diarrhea in liver transplant recipients often pose a challenge owing to the variety of infectious and non infectious causes. However, diagnosis is principally focused on ruling out an infectious etiology. Tacrolimus, an immunosuppressive agent generally used after liver transplantation, is absorbed mainly from the duodenum through the upper jejunum. It can be assumed that metabolism of the drug will be influenced by diarrhea. METHODS: Four liver transplant recipients who developed an episode of acute gastroenteritis. Infectious etiology was confirmed; trough tacrolimus levels were measured before, during and after gastroenteritis. RESULTS: All patients presented a two- to three-fold increase in blood tacrolimus levels after the onset of gastroenteritis. CONCLUSIONS: Until the role played by the intestine in the metabolism of tacrolimus is fully understood, it is prudent to recommend early dose reduction of tacrolimus and careful monitoring of trough levels during diarrheal disorders of any nature in pediatric liver-transplanted patients. PMID- 12780678 TI - The dynamics of antidonor antibody formation early after clinical kidney transplantation measured by flow cytometry and microcytotoxicity test. AB - To study the dynamics of antidonor sensitization 92 patients were monitored for antibodies against donor T and B spleen lymphocytes before transplantation, within the first month after transplantation and 3 months after transplantation. Patients were monitored for donorreactive antibodies (DRA) of immunoglobulin G (IgG), IgA, and IgM isotype using flow cytometry (FC) and the standard microcytotoxicity test (CYT). Graft function was followed for at least 2 yrs, 51% of patients for 3 yrs after transplantation. Within the first month after transplantation the percentage of patients with antidonor sensitization detected by FC rose dramatically, so that the overall sensitization rate increased from 28 (30.4%) of 92 patients prior to transplantation to 63 (68.5%) of 92 patients after transplantation. Whereas preoperatively only one isotype (IgM for T lymphocytes, IgG for B lymphocytes) and only one target cell type (either T or B lymphocytes) dominated, the postoperative patterns of positive FC results were more variable regarding target and isotype, whereby FC-DRA of the IgA class substantially contributed. Appearance of donor-directed antibodies early after kidney transplantation is a frequent event. In our cases, sensitization with FC DRA per se seemed not to be detrimental to the graft outcome (p > 0.05) but CYT DRA did resulting in a significant poor graft outcome 3 months after transplantation (p < 0.001). PMID- 12780679 TI - Hyperparathyroidism and long-term bone loss after renal transplantation. AB - BACKGROUND: While early bone loss after renal transplantation (RT) is well described, factors affecting the long-term fate of bone have received less attention. METHODS: Whole body (WB), lumbar spine (LS) and femoral neck (FN) bone mineral density (BMD) was measured using dual energy X-ray absorptionometry in 126 stable RT patients and repeated in 114 survivors after 3 yr. Percentage change per year (%/yr) was correlated to clinical and biochemical markers of bone metabolism. RESULTS: Low bone mass was a marker of increased mortality (FN < 80% normal 6.3%/yr; >80% 2.2%/yr). Percent change was WB -0.7 +/- 1.5 (p < 0.01); LS 0.3 +/- 2.6; FN -1.0 +/- 3.0 (p < 0.01) and, corrected for expected loss for age and sex: WB -0.5 (p < 0.01); LS 0.0; FN -0.8 (p < 0.05). Factors associated with increased loss rates were (LS%): short RT duration [<2 yr: -3.1 (p < 0.01)], high prednisone dose [>9 mg/d: -1.9 (p < 0.01)], high cyclosporine trough concentration [>175 ng/L: -1.9 (p < 0.05)], high hyperparathyroidism (PTH) [>150 ng/L: -1.5 (p < 0.05)], high alkaline phosphatase [>275 U/L: -1.6 (p < 0.05)], high osteocalcin [>75 microg/L: -1.6 (p < 0.05)]. Marginal detrimental effects of uremia, hypoalbuminemia and hyperphosphatemia were noted. Thiazide treatment seemed to protect against, and furosemide to exacerbate, bone loss, but this may have been related to associated uremia. Patients treated with vitamin D gained bone, while untreated patients with low initial 1,25-dihydroxyvitamin D lost bone [FN%-2.1 (p < 0.05)]. The prevalence of PTH (52%) and hypercalcemia (22%) remained unchanged. There was no effect of sex hormone levels, calcium and phosphate excretion, or serum calcium. CONCLUSION: While LS BMD stabilizes after RT, there is a continuing loss of WB and FN BMD. The major causes of bone loss are steroid therapy and continuing PTH, with no tendency towards spontaneous resolution. Increased vitamin D and calcium therapy should be considered for this patient group, and more aggressive therapy, e.g. parathyroidectomy given for patients with resistant PTH of >150 ng/L. PMID- 12780680 TI - Recurrent allograft HCV presenting as acute cellular rejection: successful management with interferon and ribavirin alone. AB - Hepatitis C virus (HCV) is currently the leading indication worldwide for orthotopic liver transplantation. However, the majority of patients receiving transplant for HCV eventually develop histopathologic evidence of recurrent allograft HCV and approximately 10% die or require retransplantation within the first 5 post-operative years because of accelerated graft injury and cirrhosis. Traditional induction immunosuppressive regimens and intensive immunosuppression used to treat episodes of acute cellular rejection (ACR) are associated with enhanced viral replication and higher likelihood and severity of recurrent HCV. At our institution, therefore, we have used low-dose steroid therapy in an effort to limit HCV replication. However, this practice has been associated with frequent early presentations consistent with ACR. Here, we present three cases consistent with histologic ACR treated with conventional antirejection therapy that improved transiently, but evolved rapidly to progressive HCV. A fourth patient with a similar presentation experienced dramatic improvement in aminotransferases when treated solely with interferon and ribavirin. We propose that histologic characteristics traditionally associated with ACR may, in fact, represent early recurrent HCV as both processes share common immunopathogenetic mechanisms, or alternatively, that both ACR and recurrent HCV may be present simultaneously. We conclude that in cases suggestive of ACR, careful consideration should be given to treatment for recurrent HCV in lieu of or in concert with intensive immunosuppression. PMID- 12780681 TI - Diagnostic efficiency of rejection monitoring after heart transplantation with cardiac troponin T is improved in specific patient subgroups. AB - Cardiac troponin T (cTnT) is a cardio-specific myofibrillar protein known to be elevated early after heart transplantation and during cardiac allograft rejection. cTnT determination in heart allograft recipients showed elevated levels in patients with higher degrees of graft rejection (International Society for Heart and Lung Transplantation grades >/=3A-4). Subgroup analyses revealed demographic patient characteristics markedly improving the diagnostic efficiency of cTnT measurement for rejection monitoring, including male recipient gender, recipient age <60 yr, female donor gender and donor age >/= 33 yr. The clinical utility of these parameters was confirmed by longitudinal patient data and may help to select recipients most likely to benefit from cTnT rejection surveillance. PMID- 12780682 TI - The role of p53 in the immunobiology of cutaneous squamous cell carcinoma. AB - Cutaneous squamous cell carcinoma is typically characterized by the over expression of the tumour suppressor protein p53. Considerable evidence suggests that immune competence is important in the control of cutaneous SCC. We discuss the immunobiology of p53 and its relevance to cutaneous SCC, including the potential interaction with human papillomavirus. PMID- 12780683 TI - Th2 cytokines, IgE and mast cells play a crucial role in the induction of para phenylenediamine-induced contact hypersensitivity in mice. AB - We previously reported the establishment of a mouse model system of contact hypersensitivity (CHS) to paraphenylemediamine (PPD). In order to analyse the functional contribution of Th2 cytokines, IL-4 and IL-5, in PPD induced CHS, STAT6 deficient (STAT6-/-) and wild-type control (WT) mice (C57BL/6) were immunized by the topical application of a PPD solution, and then the subsequent skin reactions were examined. Ear swelling was significantly reduced with a delayed peak response in STAT6-/- mice as compared with that of WT mice. A histological analysis showed the infiltration of both eosinophils and neutrophils in the skin of STAT6-/- mice challenged 24 h previously to significantly decrease in comparison with that in the WT mice. The expression of Th2 cytokines (IL-4, IL 5) by ELISA in the PPD-challenged skin tissue specimens as well as the IgE and IgG1 response after challenge were also profoundly reduced in the STAT6-/- mice. The adoptive transfer of the serum obtained from sensitized WT mice for the putative IgE transfer induced a peak response at 3 h and 24 h after challenge. To further investigate the role of mast cells in the induction of PPD-CHS, mast cell deficient W/Wv mice were sensitized with PPD and then were challenged. Maximal ear swelling was detected from 12 to 24 h and another small peak response was observed at 1 h in+/+mice, whereas only a small peak response at 24 h was detected in W/Wv mice. These data indicate that not only Th2 cytokines and IgE but also mast cells play an essential role in the induction of PPD-CHS. PMID- 12780684 TI - Absence of CD4+CD25+ regulatory T cells is associated with a loss of regulation leading to increased pathology in Helicobacter pylori-infected mice. AB - Helicobacter pylori induces symptomatic chronic gastritis in a subpopulation of infected individuals. The mechanism(s) determining the development and severity of pathology leading to symptoms are not fully understood. In a mouse model of H. pylori infection we analysed the influence of immunoregulatory CD4+CD25+ T cells on H. pylori colonization and gastritis. Athymic C57BL/6 nu/nu mice were reconstituted with (a) lymph node (LN) cells (b) LN cells depleted of CD25+ T cells (CD25(-) LN) or (c) not reconstituted at all. Mice were then infected orally with 3 x 10(8)H. pylori SS1 bacteria. At 2 and 6 weeks after the inoculation there was a significant (P < 0.001) reduction in H. pylori colonization in athymic mice transferred with CD25(-) LN cells compared to mice transferred with LN cells. Colonization was still reduced at 12 weeks after inoculation. Mice transferred with CD25(-) LN cells showed an earlier onset and increased severity of gastritis as compared to mice receiving LN cells. Splenic cells isolated from mice receiving CD25(-) LN cells produced the highest level of IFN-gamma on stimulation with H. pylori antigens in vitro, had a higher H. pylori specific DTH response and increased infiltration of CD4+ T cells and macrophages in the gastric mucosa. Athymic mice not transferred with T cells had persistent high H. pylori colonization and displayed a normal gastric epithelium without inflammatory cells. In conclusion, CD4+CD25+ cells reduce immunopathology in H. pylori infection, possibly by reducing the activation of IFN-gamma producing CD4+ T cells, even at the expense of a higher H. pylori load in the gastric mucosa. PMID- 12780685 TI - Evidence for intramolecular B-cell epitope spreading during experimental immunization with an immunogenic thyroglobulin peptide. AB - Thyroglobulin (Tg) is a target autoantigen in autoimmune thyroid diseases, such as Graves' disease (GD) and Hashimoto's thyroiditis. In a previous study we identified three 20mer Tg peptides bearing epitopes of autoantibodies associated with GD (TgP15, TgP26 and TgP41: sequences 2339-2358, 2471-2490 and 2651-2670 of human Tg, respectively). In the present study, we investigated the antigenicity of the above peptides in experimental immunization with Tg, the immunogenicity of antigenic peptides and the possibility of intramolecular B-cell epitope spreading during peptide immunization. For this purpose, two rabbits were injected with human Tg in CFA six times, every three weeks. Two control animals were injected only with CFA. Testing of antisera and of affinity-purified antibodies, by ELISA against the three peptides, revealed reactivity only to TgP41. This synthetic peptide was subsequently administered to two rabbits, in its free form (100 micro g in CFA six times, every two weeks). A strong serological response was developed not only against TgP41, but also to intact human and rabbit Tg. Immunization with TgP41 induced intramolecular B-cell epitope spreading, i.e. production of antibodies to sites on Tg other than that corresponding to TgP41, as revealed by immunoadsorption and competitive ELISA. Histopathological studies did not reveal any infiltration in thyroid glands. We conclude that peptide TgP41 encompasses not only an epitope of disease-associated autoantibodies, but also a dominant immunogenic epitope of experimentally induced Tg-specific antibodies, able to drive B-cell epitope spreading. PMID- 12780686 TI - Insight into antibody responses induced by plasmid or adenoviral vectors encoding thyroid peroxidase, a major thyroid autoantigen. AB - Plasmid and adenoviral vectors have been used to generate antibodies in mice that resemble human autoantibodies to the thyrotrophin receptor. No such studies, however, have been performed for thyroid peroxidase (TPO), the major autoantigen in human thyroiditis. We constructed plasmid and adenovirus vectors for in vivo expression of TPO. BALB/c mice were immunized directly by intramuscular injection of TPO-plasmid or TPO-adenovirus, as well as by subcutaneous injection of dendritic cells (DC) infected previously with TPO-adenovirus. Intramuscular TPO adenovirus induced the highest, and TPO-plasmid the lowest, TPO antibody titres. Mice injected with TPO-transfected DC developed intermediate levels. Antibodies generated by all three approaches had similar affinities (Kd approximately 10( 9)M) and recognized TPO expressed on the cell-surface. Their epitopes were analysed in competition assays using monoclonal human autoantibodies that define the TPO immunodominant region (IDR) recognized by patients with thyroid autoimmune disease. Surprisingly, high titre antibodies generated using adenovirus interacted with diverse TPO epitopes largely outside the IDR, whereas low titre antibodies induced by DNA-plasmid recognized restricted epitopes in the IDR. This inverse relationship between antibody titre and restriction to the IDR is likely to be due to epitope spreading following strong antigenic stimulation provided by the adenovirus vector. However, TPO antibody epitope spreading does not occur in Hashimoto's thyroiditis, despite high autoantibody levels. Consequently, these data support the concept that in human thyroid autoimmunity, factors besides titre must play a role in shaping an autoantibody epitopic profile. PMID- 12780687 TI - Oral administration of type II collagen suppresses pro-inflammatory mediator production by synoviocytes in rats with adjuvant arthritis. AB - The objective of this study was to investigate the effect of the oral administration of type II collagen (CII) on pro-inflammatory mediator production by synoviocytes in rats with adjuvant arthritis (AA). Sprague-Dawley rats were fed with bovine CII either before immunization with Complete Freund's adjuvant (CFA) or after initiation of arthritis. Hind paw secondary swelling was measured and synoviocytes were harvested. Sera from portal vein of oral tolerized rats were collected and in vitro synoviocytes culture or synoviocytes-Peyer's Patches (PP) cells coculture system were developed. Interleukin (IL)-1 activity was measured by a mouse thymocyte activation assayed by MTT dye reduction and tumour necrosis factor (TNF) activity was measured by an L929 cytotoxicity bioassay. Nitric oxide (NO) and malondialdehyde (MDA) levels were measured by biochemical methods. We found that feeding with CII (5, 50 and 500 micro g/kg) for 7 days before immunization significantly suppressed hind paw secondary swelling measured at day 16, 20, 24 and 28 (all P < 0.01) and pro-inflammatory mediator (IL-1, TNF, NO and MDA) production by synoviocytes (all P < 0.01) in rats with AA. Feeding with CII (5, 50 and 500 micro g/kg) for 7 days after initiation of arthritis had a similar effect. CII (1, 10, 100 micro g/ml) had no effect on IL-1 and TNF production by synoviocytes in vitro, but CII 10 micro g/ml suppressed IL-1 and TNF production by synoviocytes-PP cells coculture system (P < 0.01), which was antagonized by anti-TGF-beta antibody (10 micro g/ml) (P < 0.01). Portal serum (1 : 10) from oral tolerized rats suppressed IL-1 and TNF production by synoviocytes (P < 0.01), which was also antagonized by anti-TGF-beta antibody (10 micro g/ml) (P < 0.01). We conclude that oral administration of CII had prophylactic and therapeutic effects on AA and over-production of IL-1, TNF, NO and MDA by synoviocytes was suppressed. Bystander active suppression may be the main mechanism of oral CII in the suppression of synoviocyte function. PMID- 12780688 TI - Activation of human intraepithelial lymphocytes reduces CD3 expression. AB - The aim of this study was to examine in detail the low functional capacity of human intraepithelial lymphocytes (IELs) in response to phytohaemagglutinin (PHA) and CD3 ligation. Human IELs were extracted from jejunal mucosa obtained from patients undergoing gastric bypass operations for morbid obesity and compared to peripheral blood (PB) lymphocytes composed predominantly of CD8+ T cells. Calcium influx ([Ca2+]i) was analysed using Fura-2-loaded cells; IL-2 receptor expression was measured by immunofluorescence and flow cytometry; IL-2 binding was determined using radiolabelled IL-2; IL-2 production was quantified by ELISA; and apoptosis was detected with Apo 2.7 staining. Compared to naive PB CD8+ T lymphocytes, calcium influx by IELs was only transient with CD3 ligation and low in amplitude with PHA. IL-2 receptor expression was reduced after CD3 ligation, yet normal in numbers and affinity after PHA stimulation. Both cell types secreted similar amounts of IL-2. CD3 expression on IELs, but not PB CD8+ T cells, declined upon activation, due partly to incomplete reexpression after modulation. Little apoptosis was found. The partial activation of IELs in response to PHA and CD3 ligation, as manifested by diminished [Ca2+]i, resulted in a decline in CD3 expression. PMID- 12780689 TI - Interleukin-12 alone can not enhance the expression of the cutaneous lymphocyte associated antigen (CLA) by superantigen-stimulated T lymphocytes. AB - It has been reported that bacterial superantigens induce interleukin (IL)-12 dependent expression of the cutaneous lymphocyte associated antigen (CLA) and that this may be relevant to the association between certain skin diseases and infections including psoriasis and streptococcal tonsillitis. We have confirmed that the streptococcal pyrogenic superantigen C (SpeC) increases CLA expression by both CD4+ and CD8+ T cells when PBMCs are incubated in medium enriched with fetal calf serum (FCS). However, such an increase could not be induced in medium enriched with human serum (HS) even when recombinant IL-12 was added to the PBMCs cultures. Strikingly, CD4+ T cells incubated with SpeC in HS showed a marked reduction in CLA expression, which was not due to apoptosis. In contrast, SpeC did induce T cell proliferation and expression of CD25, CD54 and CD103 in the presence of HS indicating that the absence of SpeC induced CLA expression in HS was not due to SpeC inhibitors. Although addition of low amounts of lipopolysaccharide endotoxin (LPS) caused a highly significant increase in CLA expression in the absence of SpeC in cultures enriched with HS, a combination of LPS and SpeC did not increase CLA expression beyond that induced by LPS alone. The superantigen-induced CLA expression in FCS was partially inhibited by anti-IL 12 but not by anti-IL-18 or antibodies to transforming growth factor (TGF)-beta. It is concluded that IL-12 alone can not increase CLA expression but requires the help of other factor(s) present in FCS but not in HS. Although LPS can induce CLA expression it does not seem to be the factor that interacts with IL-12 to induce superantigen-mediated CLA expression in cultures enriched with FCS. PMID- 12780690 TI - Mucosal and systemic immune responses to plasmid protein pgp3 in patients with genital and ocular Chlamydia trachomatis infection. AB - The circulating and cervical B cell responses to Chlamydia trachomatis plasmid protein pgp3 were characterized in children and adults with ocular or genital chlamydial infection using the enzyme-linked immunospot assay (ELISPOT) and ELISA. No pgp3-specific ASCs were detected in healthy controls, but predominantly IgA ASCs were detected in UK adults with uncomplicated cervicitis or urethritis (P = 0.03, 0.019). In patients with extragenital complications or pelvic inflammatory disease a mixed response with more IgG and IgM ASCs was evident, suggesting a breach of mucosal immune compartmentalization with more extensive infection. In women with chlamydial cervicitis, ASCs secreting predominantly IgA, but also IgG, to pgp3 were present in cervix at presentation, with a frequency 30 50 times higher than blood. Cervical ASC numbers, especially IgG, fell markedly six weeks after antibiotic treatment. We detected principally IgA pgp3-specific antibody secreting cells (ASCs) in children resident in a Gambian endemic area, with a trend towards suppression of IgA responses during intense trachomatous inflammation (P = 0.06), as previously reported for other chlamydial antigens, and in keeping with the findings in genital disease. These data provide a rationale for further studies of immune responses to pgp3 in humans and animal models of chlamydia-induced disease, and its potential use in diagnostic assays and protective immunization strategies. PMID- 12780691 TI - The production of tumour necrosis factor-alpha is decreased in peripheral blood mononuclear cells from multidrug-resistant tuberculosis patients following stimulation with the 30-kDa antigen of Mycobacterium tuberculosis. AB - The clearance of intracellular bacteria requires the appropriate induction of proinflammatory cytokines and chemokines to recruit macrophages and T cells to the site of infection. In this study, we investigated the production of tumour necrosis factor (TNF)-alpha, interleukin (IL)-8 and interferon (IFN)-gamma by the peripheral blood mononuclear cells (PBMC) of patients with multidrug-resistant tuberculosis (MDR-TB) in response to in vitro stimulation with the 30-kDa antigen of Mycobacterium tuberculosis. The results were compared with those from cases of newly diagnosed TB (N-TB) and TB with treatment failure (TF-TB), and healthy tuberculin reactors (HTR). The most significantly depressed TNF-alpha levels were found in MDR-TB patients. IFN-gamma production was depressed significantly in all groups of TB patients compared with the HTR group. TNF-alpha secretion in response to the 30-kDa antigen was unchanged by coculturing with recombinant human interferon (rhIFN)-gamma, and was increased dramatically following IL-10 neutralization with an anti-human IL-10 antibody. The IL-8 levels were depressed significantly in MDR-TB patients compared with N-TB patients, but were similar to the IL-8 levels in TF-TB patients. Furthermore, rhTNF-alpha directly increased IL 8 secretion, and neutralizing antibody to TNF-alpha inhibited IL-8 production by the PBMC of MDR-TB patients that were stimulated with the 30-kDa antigen. Taken together, these data suggest that the PBMC of MDR-TB patients typically show TNF alpha depression in response to the 30-kDa antigen, and this effect is modulated by IL-10. In addition, we highlight the role of TNF-alpha in IL-8 secretion in MDR-TB patients. PMID- 12780692 TI - Specific lytic activity against mycobacterial antigens is inversely correlated with the severity of tuberculosis. AB - The ability of peripheral blood mononuclear cells (PBMC) from patients with active tuberculosis to display cytotoxic responses against autologous Mycobacterium tuberculosis (Mtb)-pulsed macrophages was evaluated. Non-MHC restricted cell-dependent lytic activity was observed in ex vivo effector cells from tuberculosis patients and was mediated mainly by CD3(+)gammadelta TCR(+) T (gammadelta T) cells bearing CD56 and/or CD16 molecules. MHC-restricted and non MHC restricted cytotoxic T cells (CTL) were differentially expanded upon stimulation with Mtb in tuberculosis patients and normal controls (N). Class-I restricted CD8(+) CTL and class-II restricted CD4(+) CTL were generated in PPD(+)N and to a lesser extent in PPD(-)N. Mtb-stimulated effector cells from tuberculosis patients became progressively non-MHC restricted CD4(-)CD8( )gammadelta T cells, while lytic activity of CD4(+) and CD8(+)CTL decreased gradually as the disease became more severe. On the other hand, target cells were lysed by ex vivo cells from tuberculosis patients through the Fas-FasL and perforin pathways. Mtb-induced CD4(+) CTL from tuberculosis patients and N controls preferentially employed the Fas-FasL mechanism. Mtb-induced CD8(+) CTL effector cells from patients used the perforin-based mechanism while cells from N controls also used the Fas-FasL pathway. While Mtb-induced gammadelta CTL from patients and PPD(-)N employed the latter mechanism cells from PPD(+)N individuals also used the perforin pathway. It can be concluded that shifts in the CTL response and the cytolytic mechanisms take place as the pulmonary involvement becomes more severe. PMID- 12780693 TI - Raised serum levels of interleukin-18 is associated with disease progression and may contribute to virological treatment failure in HIV-1-infected patients. AB - To gain further insight into the possible role of interleukin (IL)-18 in HIV-1 infection we examined serum levels of IL-18 in various clinical and immunological stages of HIV-1 infection during cross-sectional (n = 41) and longitudinal testing (n = 20) and during HAART (n = 21, 24 months follow-up). Our main findings were that HIV-1-infected patients had significantly raised IL-18 levels comparing healthy controls, particularly in those with advanced disease, that while HAART induced a marked decline in IL-18, virological treatment failure was associated with persistently raised IL-18 levels during such therapy and that our in vitro experiments showed an IL-18-mediated up-regulation of the HIV-1 coreceptor CXCR4 and the pro-apoptotic mediator TRAIL in PBMC from HIV-1-infected patients receiving HAART. HIV-1 infection appears to be characterized by persistently raised IL-18 levels and during HAART, such a pattern was associated with virological treatment failure, possibly contributing to immunodeficiency and HIV-1 replication in these patients. PMID- 12780694 TI - Circulating concentrations of soluble granzyme A and B increase during natural and experimental Plasmodium falciparum infections. AB - Release of soluble Granzymes (sGranzymes) is considered to reflect activation of cytotoxic T lymphocytes and NK cells. sGranzymes and a number of pro-inflammatory cytokines were measured in plasma of malaria patients with natural or experimentally induced Plasmodium falciparum infections. Concentrations of sGranzyme A and B, IL-10, IL-12p70 and CRP were significantly increased in African children presenting with clinical malaria; IL-10 and CRP concentrations were significantly correlated with disease severity. In nonimmune Dutch volunteers which were experimentally infected by P. falciparum-infected mosquitoes, sGranzyme A increment started 1-2 days prior to clinical symptoms and microscopically detectable parasitaemia. This coincided with increases in IFNgamma, IL-12p40 and IL-8, while sGranzyme B and IL-10 levels increased 24-48 h later. The elevation of sGranzyme A and IFNgamma in nonimmune volunteers suggests that NK cells are activated upon release of parasites by infected liver cells and subsequently during blood stage infection; thus, NK cells are likely involved innate immune human host resistance in the early phase of a malaria infection. PMID- 12780695 TI - Anti-Saccharomyces cerevisiae antibodies (ASCA) and autoimmune liver diseases. AB - Antibodies to the baker's yeast Saccharomyces cerevisiae (ASCA), recently proposed as a serological marker of Crohn's disease, have also been detected in other autoimmune disorders. The aim of this study was to determine prevalence and clinical significance of ASCA in autoimmune liver disease. The presence of IgG and IgA ASCA was evaluated using a commercially available immunoassay in 215 patients with autoimmune liver disease (primary biliary cirrhosis, PBC, 123 cases; autoimmune hepatitis, AIH, 67 cases; primary sclerosing cholangitis, PSC, 25 cases), 48 with inflammatory bowel disease and 19 healthy blood donors. Anti neutrophil cytoplasmic antibodies with the perinuclear pattern (p-ANCA) were assessed by indirect immunofluorescence in PSC patients. The main clinical and biochemical parameters between ASCA-positive and negative patients were analysed and compared. ASCA are predominant in Crohn's disease (70%); among liver patients, PSC and AMA-negative PBC show the highest ASCA prevalence (53% and 44%). In PBC ASCA correlate with higher levels of circulating IgA (P < 0.05). In PSC the detection of either ASCA or p-ANCA is neither associated with any clinical or biochemical feature, nor with an underlying inflammatory bowel disease. ASCA can not be considered an additional serological marker of autoimmune liver disease, but the possibility of detecting such a reactivity in autoimmune liver disorders should be considered; their correlation with elevated IgA in PBC suggests that ASCA may be an indirect sign of enhanced mucosal immunity; in PSC patients neither ASCA nor p-ANCA predict the occurrence of a concomitant inflammatory bowel disease. PMID- 12780696 TI - Measurement of soluble Fcgamma receptor type IIIa derived from macrophages in plasma: increase in patients with rheumatoid arthritis. AB - FcgammaRIII (CD16) is found in two alternative forms, a transmembrane FcgammaRIIIa expressed on NK cells and macrophages, and a glycosylphosphatidylinositol-linked FcgammaRIIIb present on neutrophils. Previously, we measured soluble FcgammaRIIIa (sFcgammaRIIIa) in plasma of NA(1 +, 2-) phenotyped donors with the anti-FcgammaRIII monoclonal antibody (MoAb) GRM1, which recognizes NA2-FcgammaRIIIb and FcgammaRIIIa. The level of sFcgammaRIIIa, as well as the total sFcgammaRIII (sFcgammaRIIIa plus sFcgammaRIIIb) in patients with rheumatoid arthritis (RA) was significantly higher than that in healthy controls. In this study, we measured sFcgammaRIIIa(M)(phi) in plasma with a newly developed anti-FcgammaRIII MoAb, MKGR14 (mIgM), which recognizes FcgammaRIIIa(M)(phi) specifically. From the recovery of purified sFcgammaRIIIa(M)(phi), the amount of sFcgammaRIIIa(M)(phi) present was about half that of sFcgammaRIIIa(NK), and that of sFcgammaRIIIa was about 50 times lower than that of sFcgammaRIIIb in pooled plasma from healthy NA(1 +, 2-) phenotyped donors. The level of sFcgammaRIIIa(M)(phi) in RA patients was about four times higher than that in healthy controls. In RA patients, both the sFcgammaRIIIa(M)(phi) and sFcgammaRIIIa levels were increased as proportionally as the Lansbury Index. The sFcgammaRIIIa, but not sFcgammaRIIIa(M)(phi) levels, were increased directly proportional to C-reactive protein. sFcgammaRIIIa(M)(phi) may be a novel marker of disease activity in RA. PMID- 12780697 TI - Release of endogenous anti-inflammatory complement regulators FHL-1 and factor H protects synovial fibroblasts during rheumatoid arthritis. AB - Rheumatoid arthritis is a chronic inflammatory disease of unknown aetiology predominantly affecting cells and tissues of synovial joints. Here we show that the two important complement regulators FHL-1 and factor H play a protective anti inflammatory role in rheumatoid arthritis. Expression analyses at the mRNA- and protein level show in vitro expression and secretion of both regulators by synovial fibroblasts derived from patients with rheumatoid arthritis. Similarly the two regulators are synthesized in vivo in diseased synovial tissue, and in particular synovial lining cells express high levels of FHL-1. The anti inflammatory role of these regulators in rheumatoid arthritis is highlighted by their induction with IFN-gamma and dexamethasone, whilst the pro-inflammatory cytokine TNF-alpha had no effect. Transient transfection experiments with various FHL-1/factor H promoter-luciferase reporter constructs into cells of distinct origin show independent cell and tissue specific promoter regulated transcription of these two regulators. The inducible expression, specifically of FHL-1 has physiological consequences. By binding directly to surfaces the released proteins protect cells from inflammatory damage and complement-mediated cell lysis. This study shows a novel protective and anti-inflammatory role of the two important complement regulators FHL-1 and factor H in rheumatoid arthritis and suggests a disease controlling role of the two proteins. PMID- 12780698 TI - Staphylococcal superantigens and T cell expansions in Wegener's granulomatosis. AB - In Wegener's granulomatosis (WG), a form of autoimmune systemic vasculitis, chronic carriage of Staphylococcus aureus constitutes a risk factor for the development of exacerbations. Circulating T cells in this disease are persistently activated, suggesting the presence of a chronic stimulus. A causal link between chronic carriage of S. aureus and chronic T cell activation in WG is conceivable, because S. aureus produces superantigens (SAg), which are potent T cell stimulators. Superantigenic stimulation of T cells results in expansion of T cell subsets expressing SAg-binding T cell receptor V-beta (Vbeta) chains. In the present study we hypothesized that in WG the presence of staphylococcal SAg is accompanied by expansion of SAg-reacting T cell subsets. We tested our hypothesis in a cross-sectional and a longitudinal study in which the association between seven staphylococcal SAg genes [typed by poplymerase chain reaction (PCR)], eight SAg-binding Vbeta chains and four SAg-non-binding Vbeta chains (assessed by flow cytometry) was assessed. Both studies showed that T cell expansions were present at a significantly higher rate in WG patients than in healthy individuals, but were not associated with the presence of either S. aureus or its SAg. Moreover, T cell expansions were generally of small extent, and did not appear simultaneously in both CD4 and CD8 subsets. We conclude that in WG S. aureus effects its supposed pathogenic function by a mechanism other than superantigenic T cell activation. PMID- 12780699 TI - Difference in antigenic determinant profiles between human and rat myeloperoxidase. AB - We tested whether rat and human MPO have similar antigenic determinants using 36 human MPO-ANCA positive sera, one mouse anti-rat MPO and four mouse anti-human MPO monoclonal reagents. Purified rat and human MPO were used in ELISA, with or without crossinhibition by preincubation with human MPO or irrelevant antigen in the liquid phase. Only one human MPO ANCA positive serum exhibited significant binding in rat MPO ELISA. This binding was poorly inhibited by preincubation with human MPO in the liquid phase, but was conserved after adsorption of non specific anti-rat activity in a chromatography column. Three mouse anti-human MPO IgG monoclonal antibodies did not recognize rat MPO. Only one mouse anti-human MPO IgA monoclonal antibody bound to rat MPO. This binding was poorly inhibited by preincubation with human MPO (35% at 2 micro g/ml). Conversely, the mouse anti rat MPO monoclonal did not bind human MPO. We have concluded that: (1) Most human MPO-ANCA recognize antigenic determinants on human MPO which are absent on rat MPO. Therefore, human auto-antibodies bind to epitopes which recently appeared after species evolution; (2) Inversely, the mouse anti-rat MPO monoclonal do not bind human MPO. Therefore, rat MPO epitopes have been altered during species evolution; (3) Mice injected with human MPO preferentially develop antibodies against xeno-epitopes which are not present in rodents. Therefore, human MPO may not be the best antigen to raise ANCA in animal models and (4) A comparison of the amino acid sequences of rat and human MPO may help elucidate the major antigenic epitopes. PMID- 12780701 TI - The clinical spectrum of nonbullous congenital ichthyosiform erythroderma and lamellar ichthyosis. AB - Until about 20 years ago, the term lamellar ichthyosis (LI) represented all nonbullous autosomal recessive ichthyoses except for harlequin ichthyosis and ichthyosis syndromes. Since the 1980s, nonbullous autosomal recessive ichthyoses have been divided into two major clinical entities, nonbullous congenital ichthyosiform erythroderma (NBCIE) and LI. The nature of scaling and intensity of erythroderma are important clinical features that distinguish between NBCIE and LI. However, a considerable number of cases show an intermediate phenotype between the two classic clinical features. Histologically, parakeratosis and inflammatory cell infiltration are seen more frequently in NBCIE than in LI and the stratum corneum is usually thicker in LI than in NBCIE. However, neither histopathological findings nor ultrastructural features seem to help clearly distinguish between NBCIE and LI. Mutations in any of the three known causative genes, TGM1, ALOXE3 or ALOX12B, can lead either to NBCIE or LI. Candidate genes specific to either NBCIE or LI alone have not been identified. Based on these facts, it might be better to consider NBCIE and LI as variations of a single keratinization disorder, although the classification of these autosomal recessive congenital ichthyosis patients into NBCIE or LI depending on their clinical features is still useful for practical patient management. PMID- 12780700 TI - Beta-2-glycoprotein I expression on monocytes is increased in anti-phospholipid antibody syndrome and correlates with tissue factor expression. AB - It is well known that monocytes may play an active role in thrombogenesis, since they may express on their surface tissue factor, the major initiator of the clotting cascade. The results of this investigation demonstrate beta-2 glycoprotein I (beta2-GPI) mRNA expression by human peripheral blood monocytes, indicating that these cells synthesize beta2-GPI. In addition, we show beta2-GPI expression on cell surface of these cells by flow cytometric analysis, and the presence of this protein in cell lysate by Western blot. Interestingly, beta2-GPI expression on monocytes is significantly increased in patients with anti phospholipid syndrome (APS) or systemic lupus erythematosus (SLE) as against healthy blood donors and correlates with tissue factor expression on monocytes. These findings support the view that monocytes are able to synthesize beta2-GPI and suggest that patients with APS may have increased beta2-GPI exposure on cell surface, which leads to persistently high monocyte tissue factor expression and consequently to a prothrombotic diathesis. PMID- 12780702 TI - Treatment of persistent painful nodules of hidradenitis suppurativa with cryotherapy. AB - We report outcomes for 10 patients with persistent painful nodules of hidradenitis suppurativa treated with cryotherapy. Eight patients reported improvement and to date have had no recurrence of lesions at the treated sites. Most patients had significant pain during and after treatment. Eight patients had post-treatment ulceration, infection or both. The average number of days for the treatment areas to heal was 25. Seven patients rated cryotherapy as better than oral antibiotics and eight patients would consider this treatment again in the future. Cryotherapy can be an effective treatment for patients who have limited but persistent painful nodules. However, patients must be warned about pain, prolonged healing time and risk of infection after the procedure. PMID- 12780703 TI - Chronic macrocheilia: a clinico-pathological study of 28 patients. AB - Chronic macrocheilia has a multifactorial aetiology and is often a diagnostic and therapeutic challenge. Epidemiological information on this condition is scarce, most of the data reported relating only to granulomatous cheilitis. We have performed a detailed clinico-pathological analysis of all patients with chronic macrocheilia presenting to us during the last 6.5 years. Of the 28 patients identified, 13 (46.4%) had granulomatous cheilitis (GC), six (21.4%) had tuberculosis of the lip, three (10.7%) had leprous macrocheilia, two (7.1%) had multiple endocrine neoplasia type IIb, and one each had Ascher's syndrome and non Hodgkin's lymphoma. Two patients were diagnosed as 'nonspecific cheilitis'. Histopathological differentiation between tuberculosis and GC was often not possible; but PCR for Mycobacterium tuberculosis was positive in all patients with tuberculosis and negative in four patients with GC in whom M. tuberculosis was sought. In spite of detailed clinical examination and investigations, a therapeutic trial was required to confirm the diagnosis in five (17.9%) patients. We have reviewed the available literature on this subject, and to our knowledge this study is the first of its kind. More such studies from other centres will help physicians to make an accurate aetiological diagnosis and treat this uncommon but disfiguring condition with confidence. PMID- 12780704 TI - Post-surgical zygomycotic necrotizing subcutaneous infection caused by Absidia corymbifera. AB - Necrotizing infections of the skin and subcutaneous tissue are usually bacterial in origin. Rarely, fungi of the class Zygomycetes, which cause deep mycoses, may be responsible for acute necrotizing infections of these areas. Several local and systemic predisposing factors have been associated with such acute necrotizing infections. Surgical debridement and amphotericin B remain the mainstay of treatment. In this report we describe a patient with post-surgical necrotizing subcutaneous infection caused by Absidia corymbifera, 2 weeks following appendectomy. Successful management with surgical debridement, topical amphotericin B and oral potassium iodide is reported. PMID- 12780705 TI - Severe generalized argyria secondary to ingestion of colloidal silver protein. AB - Argyria is a rare cause of cutaneous discolouration caused by silver deposition. We report a case of dramatic and diffuse argyria secondary to ingestion of colloidal silver protein over a 1-year period. Stained electron microscopy with spectral analysis was used to confirm the clinical diagnosis. Silver-protein complexes are deposited in the skin and reduced to inert silver salts by sunlight in a process similar to that harnessed in photography. Our patient had obtained the silver for consumption via mail order. It had been advertised as a cure for a variety of diseases. Colloidal silver protein is commercially available as a 'food supplement', hence circumventing the strict controls placed on medicines. PMID- 12780706 TI - Giant lichenification of the scalp. AB - Lichenification is characterized clinically by thickening of areas of skin as a result of the itch-scratch cycle and therefore is seen in conditions associated with chronic pruritus. The characteristic feature of giant lichenification is the occurrence of tumour-like growths with a warty cribriform surface. We describe a renal transplant patient presenting with giant lichenification of the scalp following an attack of herpes zoster at the same site. Chronic pruritus following scalp dysaethesia secondary to herpes zoster was considered the most likely explanation for the occurrence of these lesions. PMID- 12780707 TI - Successful treatment of multiple filiform facial warts with imiquimod 5% cream in a patient infected by human immunodeficiency virus. AB - Imiquimod, an imidazoquinoline amine, is approved for the topical treatment of external anogenital warts induced by human papilloma virus. Several clinical trials have shown imiquimod to be an effective and safe drug for treatment of anogenital warts. Consequently, it was considered that imiquimod might be effective on warts caused by the same aetiological agent located on other skin areas. We describe the favourable outcome of a case of multiple facial verrucae in a human immunodeficiency virus (HIV)-infected patient treated with imiquimod 5% cream. This is a promising finding which supports those of two previous reports. We feel that imiquimod could be used in HIV-infected patients with multiple facial warts in whom conventional therapies are ineffective or produce significant side-effects. PMID- 12780708 TI - Acquired idiopathic generalized anhidrosis: a rare cause of heat intolerance. AB - A reduced level or complete cessation of sweating can be caused by a variety of factors affecting sweat glands directly or indirectly through alterations in their nerve supply. The most common presentation is the syndrome of heat intolerance with or without features of dysautonomia. An acquired idiopathic form of generalized anhidrosis is characterized by loss of sweating in the absence of any neurological features or destruction of sweat glands. Cholinergic urticaria has been associated with some cases of this acquired idiopathic form of generalized anhidrosis. A case of generalized absence of sweating without a neurological deficit is described, and the implications of heat intolerance are discussed. PMID- 12780709 TI - Chronic actinic dermatitis as the presenting feature of HIV infection in three Chinese males. AB - There have been a few reports in the literature of chronic actinic dermatitis (CAD) associated with HIV infection, mostly in African--Americans of skin type VI, where photosensitivity predated the diagnosis of HIV infection. We report three cases, all Chinese males with skin type III or IV, who presented to our centre with CAD, and in whom advanced asymptomatic HIV infection was subsequently diagnosed. All had CD4 cell counts less than 100 cells/ micro L, with no evidence of AIDS-related complex. They were treated conservatively with photoprotection and topical steroids with mild to moderate improvement. A comparison with nine previously reported cases is made. The pathogenesis of CAD is unclear, but predominance of CD8 cells in severe cases and reversal of the CD4 : CD8 ratio in lesional skin and peripheral blood of HIV-negative CAD patients has been observed. CAD may be consequent to, and a presenting feature of, advanced HIV infection. PMID- 12780710 TI - Treatment of toxic epidermal necrolysis in AIDS with intravenous immunoglobulins. AB - Individuals with AIDS are at higher risk of developing severe cutaneous adverse drug reactions. We report two AIDS patients with drug-induced toxic epidermal necrolysis (TEN). The suspected drugs were discontinued. Both patients were treated with intravenous human immunoglobulins at a dose of 1 g/kg body weight per day for two consecutive days and both experienced a good outcome. Intravenous immunoglobulin potentially lowers the morbidity and mortality of TEN and shortens the duration of the patient's hospitalization. PMID- 12780711 TI - Lymphomatoid contact dermatitis to para-tertyl-butyl phenol resin. AB - Lymphomatoid contact dermatitis refers to the relatively little known phenomenon of allergic contact dermatitis producing histological features suggestive of cutaneous T-cell lymphoma. We report the first case of lymphomatoid contact dermatitis in response to para-tertyl-butyl phenol resin. PMID- 12780712 TI - Pyoderma gangrenosum in a thoracotomy wound associated with a pulmonary cavitating lesion. AB - We present a patient with both pyoderma gangrenosum in a thoracotomy wound and a pulmonary cavitating lesion with the histological features of Wegener's granulomatosis. An ulcer with blue undermined edges developed in our patient's thoracotomy scar after a lobectomy was carried out for the cavitating lung lesion and c Antineutrophil Cytoplasmic Antibodies (cANCA), which is highly specific for active Wegener's granulomatosis but was negative at the time of her skin lesion. This suggests that our patient had cutaneous pyoderma gangrenosum rather than cutaneous Wegener's granulomatosis, although the two conditions may occasionally have similar clinical and histological features. PMID- 12780713 TI - Papuloerythroderma of Ofuji associated with acute myeloid leukaemia. AB - Papuloerythroderma of Ofuji (PEO) is an uncommon skin condition most commonly described in Japan. The aetiology of PEO is unknown and treatment may be difficult. There are several reports of an association between PEO and malignancy, most particularly with T-and B-cell lymphomas. We describe PEO complicated by acute myeloid leukaemia occurring in a 62-year-old man. This association has not been reported previously. PMID- 12780714 TI - Nonsynchronized segmented heterochromia in black scalp hair. AB - Nonsynchronized segmented heterochromia in black scalp hair is a rarely reported entity, the only previous report being described in association with iron deficiency anaemia. A 14-year-old girl presented with a 2-year history of nonsynchronized segmented heterochromia. She was otherwise well and her serum iron, copper, zinc and protein levels were all within the normal range. She had no clinical evidence of vitiligo or alopecia areata. This patient is believed to represent the first reported case of nonsynchronized segmented heterochromia in black scalp hair as a presentation of premature greying of the hair. PMID- 12780715 TI - Onychomycosis due to Onychocola canadensis. AB - Onychocola canadensis is an uncommon nondermatophyte mould which can occasionally cause onychomycosis, and rarely, tinea manuum. It typically affects elderly individuals, some of whom have been farmers, in temperate climates. We describe a patient with onychomycosis due to this organism. She failed to respond to a 12 week course of terbinafine 250 mg o.d. PMID- 12780716 TI - Effectiveness of oral Ginkgo biloba in treating limited, slowly spreading vitiligo. AB - For effective treatment of vitiligo, it is as important to arrest the progression of the disease as it is to induce repigmentation. Recently, oxidative stress has been shown to play an important role in the pathogenesis of vitiligo. Ginkgo biloba extract has been shown to have antioxidant and immunomodulatory properties. In a double-blind placebo-controlled trial, we evaluated the efficacy of G. biloba extract in controlling the activity of the disease process in patients with limited and slow-spreading vitiligo and in inducing repigmentation of vitiliginous areas. Fifty-two patients were assigned to two treatment groups (A and B) in a double-blind fashion, but only 47 patients could be evaluated, because one patient in group A and four patients in group B withdrew for reasons unrelated to the study. Patients in group A were given G. biloba extract 40 mg three times daily whereas patients in group B received placebo in similar doses. A statistically significant cessation of active progression of depigmentation was noted in patients treated with G. biloba (P = 0.006). Marked to complete repigmentation was seen in 10 patients in group A, whereas only two patients in group B showed similar repigmentation. The G. biloba extract was well tolerated. G. biloba extract seems to be a simple, safe and fairly effective therapy for arresting the progression of the disease. PMID- 12780717 TI - Nitric oxide and cellular immunity in experimental cutaneous leishmaniasis. AB - We examined the local and systemic production of nitric oxide (NO) and the pattern of cytokine during the course of Leishmania mexicana infection in susceptible BALB/c and resistant C57BL/6 mice. NO derivatives were measured in serum, and the expression of inducible nitric oxide synthase (iNOS), interferon (IFN-gamma), interleukin (IL-4) and epidermal Langerhans cells (LC) was measured in the lesions by immunohistology. Circulating NO concentrations, iNOS+ cell density, IFN-gamma+ Th1 cells and CD205+ Langerhans cells were higher in early lesions of resistant C57BL/6 mice. In contrast, susceptible BALB/c mice developed chronic and progressive lesions with a predominance of IL-4+ Th2 cells. In both susceptible and resistant mice, lesion size and lymph node volume followed a similar course. The early local and systemic production of NO in resistant mice may be related with the premature production of IFN-gamma observed, contributing to the resolution of the lesion. PMID- 12780718 TI - Selenium protects primary human keratinocytes from apoptosis induced by exposure to ultraviolet radiation. AB - The generation of reactive oxygen species has been implicated in ultraviolet radiation (UVR)-induced skin damage. In mice, increasing dietary selenium intake protects skin from UVR-induced DNA damage and photocarcinogenesis. We sought to determine whether selenium supplementation could protect keratinocytes from apoptosis resulting from exposure to broadband (TL20W/12) UVR. Unirradiated cultures contained 6.5 +/- 1% apoptotic cells; the maximum percentage of apoptotic cells (34 +/- 5%) was seen 16 h after UVR of 600 J/m(2). Under these conditions cell death from necrosis was 15 +/- 2.5% of the total cells. A 24-h preincubation with sodium selenite (10 nm(-1) microm) or selenomethionine (50 nm( 1) microm) protected cultured human keratinocytes from UVR-induced apoptosis. In primary keratinocytes the greatest reduction in apoptosis was found with 100 nm of either selenium compound (71% reduction in the numbers of total apoptotic cells; P < 0.01). Supplementation with 100-200 nm selenite or selenomethionine prevented UVR-induced apoptosis, but did not decrease the levels of UVR-induced p53, as measured by Western blotting. Collectively, this data suggests that selenium prevents UVR-induced cell death by inhibiting p53-independent cell death pathways. PMID- 12780719 TI - Clinicopathological features and HLA tissue typing in pemphigoid gestationis patients in Kuwait. AB - Pemphigoid gestationis (PG) is a rare autoimmune disease of pregnancy. We report a series of 22 cases of PG in Kuwait. They constituted 18% of all the autoimmune bullous diseases registered in our centre over a span of 11 years. PG was observed to be the third most common bullous disease in our region. Ninety-five per cent of the patients were of Arab ethnicity. The clinical features observed in our patients were comparable to those reported elsewhere. Systemic steroids (prednisolone 20-60 mg daily) remained the mainstay of treatment to control the active disease and an optimal dose of 20 mg of prednisolone was maintained throughout the pregnancy and immediate postpartum period. We observed a favourable outcome of pregnancies complicated by PG without any associated maternal or foetal morbidity. Kuwaiti patients with PG were observed to have a predominance of HLA-DR3 and DQ2 antigens. No predominance of HLA-DR4 antigen was observed. PMID- 12780720 TI - Association between long-term acitretin therapy and osteoporosis: no evidence of increased risk. AB - Osteoporosis has been observed with chronic hypervitaminosis A, leading some authors to hypothesize that systemic retinoids may have an effect on bone mineral density. Two previous small studies identified osteoporosis in patients who received long-term therapy with etretinate. Etretinate has now been superceeded by acitretin in clinical use. We hypothesized that bone mineral density is lower in patients taking long-term acitretin than control cases who had never taken acitretin. Thirty Caucasian patients receiving acitretin for a median of 3.6 years for a variety of dermatoses were studied. Bone mineral density measurements were determined using DEXA scanning at two standard sites, the lumbar spine and Ward's triangle. We did not find an association between daily dose of acitretin, total dose administered or overall duration of treatment and risk of osteopenia or osteoporosis. Acitretin appears to be safe for long-term use in patients with chronic dermatoses. PMID- 12780721 TI - Cytomorphological variations, proliferation and angiogenesis in the prognosis of cutaneous melanoma. AB - Depth of invasion and stage of the disease are well known prognostic indicators in cutaneous melanoma (CM). However, the role of other parameters, such as the variations in cytomorphology of melanocyte tumours, mitotic activity and angiogenesis is still open to question. The aim of this study was to analyse proliferation by mitotic activity index (MAI) and immunostaining of proliferating cell nuclear antigen (PCNA), and the intensity of neovascularization (microvessel density; MVD) in CM clinical stage I in relation to epithelioid, spindle and nevoid cell type, histological type (superficial spreading melanoma and nodular melanoma), Clark's level and Breslow thickness. Finally, the role of all parameters in the prognosis of CM was evaluated. Statistical analysis demonstrated that cytological characteristics of CM correlate only with Clark's level, while histological types correlate with MAI, PCNA and MVD. MAI and PCNA also showed correlation between groups according to Clark's level and Breslow thickness. Finally, tumour cell PCNA was found to correlate with MVD. Survival of patients with CM correlated significantly with MAI. These results suggest that cytological variation, histological type, PCNA and MVD alone are not independent prognostic parameters, whereas MAI is a potentially important prognostic marker in CM. PMID- 12780722 TI - Cutaneous endometriosis: non-invasive analysis by epiluminescence microscopy. AB - The clinical appearance of cutaneous endometriosis can share some features with malignant melanoma, thus representing a possible cause for concern in both patient and clinician. In recent years, the use of epiluminescence microscopy (ELM, dermoscopy) has proved useful in improving the accuracy of diagnosis of pigmented skin lesions. The purpose of this study was to analyse the dermoscopic features of cutaneous endometriosis with histopathological correlation. We studied a case which showed homogeneous reddish pigmentation, regularly distributed. Within this typical pigmentation there were small red globular structures, but more defined and of a deeper hue, which we called 'red atolls'. ELM thus revealed a distinctive pattern in cutaneous endometriosis. PMID- 12780723 TI - Over-expression of Mn-superoxide dismutase as a marker of oxidative stress in lesional skin of chronic idiopathic urticaria. AB - We studied the involvement of oxidative stress in chronic idiopathic urticaria (CIU), assessing the activities of superoxide dismutase (SOD) and glutathione and the levels of malondialdeyde (MDA), a marker of lipid peroxidation, in samples taken from lesional skin (n = 16) and nonlesional skin (n = 11) of CIU patients. The activity of SOD and glutathione and the levels of MDA were markedly increased in lesional skin as compared with skin of healthy subjects, whereas no differences were detected between nonlesional skin of CIU patients and control samples. Immuno-dot blot assay revealed an up-regulation of Mn-SOD expression in lesional skin. These findings show that oxidative stress is crucially involved in CIU. The evidence of lipid peroxidation and compensatory increase of Mn-SOD and glutathione activities in lesional skin, in the absence of any alteration in uninvolved skin, suggests that oxidative stress is secondary to the development of inflammation. PMID- 12780724 TI - Xanthoma striatum palmare as an early sign of familial type III hyperlipoproteinemia with an apoprotein E genotype epsilon2/epsilon2. PMID- 12780725 TI - Eczema in a port-wine stain. PMID- 12780726 TI - Connective tissue naevus (collagenoma) in a patient with benign joint hypermobility syndrome (Ehlers-Danlos syndrome type III). PMID- 12780727 TI - Recurrent confluent and reticulate papillomatosis of Gougerot and Carteaud in siblings. PMID- 12780728 TI - Acral variant of eosinophilic pustular folliculitis (Ofuji disease). PMID- 12780729 TI - Childhood pemphigus associated with montelukast administration. PMID- 12780730 TI - Punctate porokeratosis in a renal transplant recipient. PMID- 12780731 TI - Aleukaemic leukaemia cutis in a patient with acute T-cell lymphoblastic leukaemia. PMID- 12780732 TI - "Tinea pseudoimbricata": tinea corporis in a renal transplant recipient mimicking the concentric rings of tinea imbricata. PMID- 12780733 TI - Delayed diagnosis of angiotensin-converting enzyme (ACE) inhibitor induced angioedema and urticaria. PMID- 12780734 TI - Case 1: enlarging lesion on right inner thigh. Diagnosis: erythema chronicum migrans caused by Lyme disease. PMID- 12780735 TI - Case 2: sudden ecchymosis of the legs with feelings of pain and weakness. Diagnosis: adult scurvy. PMID- 12780736 TI - Case 3: widespread scaly papules affecting a patient's limbs and pinnae giving the skin a rough feeling. Diagnosis: Flegel's disease. Syn. hyperkeratosis lenticularis perstans. PMID- 12780737 TI - Case 4: eruption on the face of a diabetic man suffering from retinopathy, hypertension, and nephropathy. Diagnosis: ciclosporin-associated hyperplastic folliculitis. PMID- 12780738 TI - Case 5: extensive skin eruption causing skin to become tight, sore, and itchy. Diagnosis: scleromyxoedema. PMID- 12780739 TI - Case 6: plaques extending in a linear pattern from left ankle to hip forming over a 2-year period. Diagnosis: linear porokeratosis with disseminated superficial porokeratosis erupting in pregnancy. PMID- 12780741 TI - Pituitary tumour transforming gene in endocrine cancer. PMID- 12780742 TI - Hyperprolactinaemia due to macroprolactins: some progress but still a problem. PMID- 12780743 TI - A practical method for the detection of macroprolactinaemia using ultrafiltration. AB - OBJECTIVE: The aim of this study was to evaluate a method for the estimation of the percentage of big-big prolactin (bb-PRL) in serum, based on centrifugal ultrafiltration (UF) and to be used for the detection of macroprolactinaemia. DESIGN: Serum samples were submitted to gel filtration chromatography (GFC), UF and precipitation with polyethylene glycol (PEG). PATIENTS: Serum samples obtained from 13 patients with macroprolactinaemia (Group 1) and from 15 hyperprolactinaemic patients without macroprolactinaemia (Group 2) were studied by the three methods. MEASUREMENTS: PRL was measured in serum, fractions from GFC, filtrate and retentate obtained after UF (this last measurement was used only to study the 'recovery' of PRL after UF and was not necessary for clinical purposes), and supernatant obtained after PEG precipitation. The calculation of the percentage of bb-PRL, by UF, was made using a simple algorithm. RESULTS: In Group 1 the percentages (mean and SD) of bb-PRL as determined by GFC, UF and PEG were, respectively, 73 +/- 16, 83 +/- 5.3 and 78 +/- 7.0. In Group 2, the percentages of bb-PRL were, in the same order as above, 4.3 +/- 8.2, 16 +/- 19 and 3.7 +/- 11. The difference of the means between Groups 1 and 2 was highly significant (P << 0.001) for all the three methods. In Group 1 the percentages of bb-PRL were widely scattered when measured by GFC but closely clustered when measured by either UF or PEG. The opposite occurred in Group 2. CONCLUSIONS: The centrifugal ultrafiltration method for the detection of macroprolactinaemia is simple, inexpensive, requires only two prolactin assays and yields the same results as the polyethylene glycol precipitation method. PMID- 12780744 TI - Bone mineral density outcomes following long-term treatment with subcutaneous testosterone pellet implants in male hypogonadism. AB - BACKGROUND: Osteoporosis is a common complication of untreated male hypogonadism. Even mild hypogonadism due to suboptimal testosterone replacement may result in decreased bone mineralization and osteoporosis. OBJECTIVE: To assess bone mineral density in hypogonadal men following long-term long-acting subcutaneous testosterone pellet implants as replacement therapy. PATIENTS: A cross-sectional study of 37 patients with primary or secondary hypogonadism receiving long-term (mean 6.6 years) subcutaneous testosterone pellet implants as replacement therapy. MEASUREMENTS: Bone mineral density was measured in all patients using dual energy X-ray absorptiometry. Serum testosterone 3-4 months after insertion of pellets was measured in all patients to assess adequacy of replacement therapy. RESULTS: Mean areal bone mineral density were 1.02 (SD 0.14) g/cm2 with a mean Z score of -0.64 (SD 1.3) and 0.87 (SD 0.13) g/cm2 with a mean Z score of 0.72 (SD 1.2) at lumbar spine and neck of femur, respectively. Mean serum testosterone 3-4 months after pellets insertion was 15.45 nmol/l (SD 4.2 nmol/l). There was no significant correlation between bone mineral density and patient's age at start or duration of testosterone therapy. CONCLUSIONS: Bone mineral density in long-term regularly treated hypogonadal men was not different from the age-matched reference range for normal men. Long-acting subcutaneous testosterone pellet implants as replacement therapy in male hypogonadism are safe, acceptable to the patient, result in adequate bone mass accumulation and maintenance of normal bone mineral density. By provision of sustained physiological levels of testosterone they may contribute to increased androgen effect at the receptor level. PMID- 12780745 TI - The correlation between the response to progestogen treatment and the expression of progesterone receptor B and 17beta-hydroxysteroid dehydrogenase type 2 in human endometrial carcinoma. AB - OBJECTIVE: In situ metabolism and synthesis of oestrogens are considered to play important roles in the pathogenesis and development of human endometrial endometrioid adenocarcinoma. Approximately 3-5% of patients with these neoplasms are under age 40, some of whom have been treated with progestogen alone as a primary therapy for both atypical endometrial hyperplasia and adenocarcinoma in order to preserve their fertility. Medroxyprogesterone acetate (MPA) has been used extensively in the treatment of both breast and endometrial disorders as an endocrine therapy. However, details of the alterations of in situ oestrogen metabolism following progestogen treatment have yet to be fully elucidated. DESIGN, PATIENTS AND MEASUREMENTS: In this study we examined the immunolocalization of 17beta-hydroxysteroid dehydrogenase (17beta-HSD) types 1 and 2, oestrogen receptor (ER), progesterone receptor (PR)A + PRB, PRB, and Ki67 in progestogen-treated endometrial endometrioid adenocarcinoma (16 cases). We compared our findings both prior to and following treatment. These findings were then correlated with the treatment outcome of individual patients in order to elucidate factors associated with the response to treatment. RESULTS: 17beta-HSD type 2 immunoreactivity was detected in 8/16 cases examined, whereas 17beta-HSD type 1 immunoreactivity was undetected in all cases examined. 17beta-HSD type 2 positive immunostaining, PRA + PRB labelling index (LI), and PRB/PRA + PRB ratio were all significantly higher in cases responding to the treatment than in those not responding. There were no significant correlations between responsive and nonresponsive cases for positive 17beta-HSD type 1 immunostaining, Ki67 LI, ER LI and age. There were no significant differences in the positive immunostaining for 17beta-HSD types 1 and 2, Ki67 LI, ER LI, PRA + PRB LI, age and PRB/PRA + PRB ratio between specimens taken prior to and following progestogen treatment. CONCLUSION: These results suggest that in situ abundance of 17beta-HSD type 2 and PR, especially PRB, can predict the possible response of patients with endometrial carcinoma to progestogen treatment. PMID- 12780746 TI - The effects of soy protein containing isoflavones on lipids and indices of bone resorption in postmenopausal women. AB - OBJECTIVE: To assess the effect of a dietary soy protein supplement containing isoflavones on lipids and indices of bone resorption in postmenopausal women. DESIGN: Placebo-controlled, double-blind, randomized study. PATIENTS: One hundred and six postmenopausal women were randomized to dietary soy supplementation (n = 51) or placebo (n = 55) for 3 months, of which 78 were included in the final analysis. MEASUREMENTS: Lipid profiles including total, low-density lipoprotein (LDL) and HDL cholesterol as well as triacylglycerol were measured. Pyridinoline and deoxypyridinoline were used as markers of bone resorption. Urinary isoflavone excretion was measured to assess compliance. RESULTS: There was a significantly greater increase in urinary isoflavone excretion detected in the soy group compared to placebo. Lipid profiles improved with significant decreases in LDL cholesterol (-0.60 +/- 0.10 vs.-0.29 +/- 0.09 mmol/l, P < 0.05), triacylglycerol (-0.22 +/- 0.07 vs. +0.01 +/- 0.05 mmol/l, P < 0.005) and the LDL : HDL ratio ( 0.32 +/- 0.10 vs. +0.20 +/- 0.10, P < 0.005) in the soy group compared to placebo. There were no significant differences between the soy and placebo groups for urinary excretion of pyridinoline (-3.8 +/- 3.1 vs.-0.8 +/- 3.1 nmol/mmolCr, P = 0.4) or deoxypyridinoline (-0.8 +/- 0.9 vs.-0.3 +/- 0.7 nmol/mmolCr, P = 0.4). CONCLUSIONS: In postmenopausal women, dietary supplementation with soy protein containing isoflavones does not appear to have oestrogenic effects on markers of bone resorption. Soy protein favourably affected lipids; however, these effects (fall in triacylglycerol and no change in HDL) differ from those observed with oral oestrogen. These findings suggest that soy may not have biologically significant oestrogenic effects on bone resorption and we hypothesize that the lipid effects may be mediated, at least in part, through nonoestrogenic mechanisms. PMID- 12780747 TI - Diurnal rhythms of serum total, free and bioavailable testosterone and of SHBG in middle-aged men compared with those in young men. AB - BACKGROUND: Conflicting views are reported on the association between advancing age and gradually diminishing concentrations of serum total testosterone in men. The putative loss of diurnal rhythm in serum total testosterone in older men is reported to be in part due to low concentrations in the morning when compared to concentrations found in young men. We have measured total, free and bioavailable testosterone along with SHBG in samples taken every 30 min throughout a 24-h period in 10 young and eight middle-aged men. RESULTS: Both young and middle-aged men displayed a significant diurnal rhythm in all variables, with a minimum fall of 43% in total testosterone from peak to nadir in all subjects. Subjecting the data to a time series analysis by least squares estimation revealed no significant difference in mesor (P = 0.306), amplitude (P = 0.061) or acrophase (P = 0.972) for total testosterone between the two groups. Comparing bioavailable testosterone in the two groups revealed no significant difference in mesor (P = 0.175) or acrophase (P = 0.978) but a significant difference (P = 0.031) in amplitude. Both groups display a significant circadian rhythm (middle-aged group P < 0.001; young group P = 0.014). Free testosterone revealed a highly significant rhythm in both the young group (P < 0.001) and the middle-aged group (P = 0.002), with no significant difference between the groups in mesor (P = 0.094) or acrophase (P = 0.698). Although analysis of the SHBG data revealed a significant rhythm in the young group (P = 0.003) and the older group (P < 0.001), the acrophase occurred in the mid afternoon in both groups (15.12 h in the young and 15.40 h in the middle-aged). The older men had a significantly greater amplitude (P = 0.044) but again no significant difference was seen in mesor (P = 0.083) or acrophase (P = 0.477) between the two groups. Acrophases for total, bioavailable and free testosterone occurred between 07.00 h and 07.30 h; for SHBG the acrophase occurred at 15.12 h in the young group and 15.40 h in the middle-aged group. CONCLUSIONS: The study suggests that the diurnal rhythm in these indices of androgen status is maintained in fit, healthy men into the 7th decade of life. PMID- 12780748 TI - The corticotrophin-releasing hormone test is the most reliable noninvasive method to differentiate pituitary from ectopic ACTH secretion in Cushing's syndrome. AB - OBJECTIVE: It has been reported previously that the paired interpretation of the corticotrophin-releasing hormone (CRH) test and the 8-mg dexamethasone suppression test (HDDST) could have higher diagnostic power than any single test in the differential diagnosis of ACTH-dependent Cushing's syndrome. This finding has not been confirmed thereafter in large series. The aim of the present study has been to assess the operating characteristics of either the CRH test or the overnight HDDST and also to evaluate the potential utility of combining the interpretation of both tests in the differential diagnosis of ACTH-dependent Cushing's syndrome. DESIGN AND PATIENTS: We have reviewed the medical records of 59 consecutive cases with ACTH-dependent Cushing's syndrome: 49 patients with proven Cushing's disease (CD) and 10 patients with proven ectopic ACTH syndrome (EAS). Univariate curves of the receiver operating characteristics (ROC) have been performed to define the best cut-off values, the sensitivity and the specificity for CRH and overnight HDDST. A comparison between the areas under the ROC curves has also been performed. RESULTS: For the CRH test, the point on the ROC curve closest to 1 corresponded to a value of ACTH percentage increment of 50%[sensitivity 86% (72.6-94.8) and specificity 90% (55.5-98.3)]. The best threshold for cortisol percentage (30%) increment gave inferior results [sensitivity 61% (45.5-75.6) and specificity 70% (34.8-93.0)]. For the HDDST, the point on the ROC curve closest to 1 corresponded to a value of cortisol decrease from the baseline of 50%[sensitivity 77% (62.7-88.5), specificity 60% (26.4 87.6)]. The area under the ROC curve of the ACTH percentage increment after CRH was significantly greater than the area under the diagonal [0.9 (0.7-1.0), P= 0.0001]. Conversely, the area under the cortisol percentage decrement after dexamethasone was not different from that obtained by chance [0.7 (0.5-0.9), P= ns]. The area under the ROC curve of CRH is significantly greater than that of overnight HDDST (P = 0.03). A correct diagnosis has been achieved by the CRH test in 86.5% of cases and by the HDDST in 73% (P = 0.06). The combination of both tests has given a correct diagnosis in a significantly lower percentage of cases than the CRH test alone (69%, P= 0.04). The bilateral inferior petrosal sinus sampling (BIPSS) has been performed in 29 patients (24 CD, five EAS) who had negative imaging and/or discordant results of the noninvasive tests. Considering the criterion of a central to peripheral ACTH ratio > 3 after CRH stimulation, a correct diagnosis was achieved in all cases. CONCLUSIONS: The present data suggest that the CRH is likely to be the most reliable noninvasive diagnostic procedure for the differential diagnosis of the ACTH-dependent Cushing's syndrome. The criterion for a diagnosis of EAS is an ACTH percentage increment lower than 50%. The use of a combination of tests is not recommended because it does not add valuable information and may even impair the outcome of the CRH test. Cases with discordant results in pituitary imaging and CRH test should undergo BIPSS. The validity of this approach, which is straightforward and easily applicable in clinical practice, should be verified in larger series. PMID- 12780749 TI - Bone mineral density in acromegaly: the effect of gender, disease activity and gonadal status. AB - OBJECTIVE: Data on bone mineral density (BMD) in acromegaly are conflicting as most previous studies collectively evaluated eugonadal and hypogonadal patients of both sexes, with or without active disease. We have evaluated BMD in 152 acromegalic patients of both sexes with varying disease activity and gonadal status. DESIGN: Cross-sectional, retrospective. PATIENTS: We studied 152 acromegalic patients (99 women aged 26-72 years, and 53 men aged 21-75 years), 107 with active and 45 with controlled disease. Eighty-five patients had normal gonadal status and 67 were hypogonadal. MEASUREMENTS: In all patients we measured serum GH levels by immunoenzimometric assay, and serum IGF-I levels by radioimmunoassay. BMD was assessed at spine L2-L4 (LS) and at femoral neck (FN) by dual energy X-ray absorptiometry; results are expressed as Z-values. RESULTS: We evaluated the effect of GH excess on bone at different sites in relation to gonadal status, disease activity and gender. At LS, in respect to the reference population, BMD (mean +/- SE) values were higher in eugonadal patients (active: 0.71 +/- 0.29, P < 0.02; controlled: 0.65 +/- 0.28, P < 0.05) and lower in hypogonadal ones (active: -0.64 +/- 0.35, 0.1 < P < 0.05; controlled: -1.05 +/- 0.36, P < 0.01), regardless of disease activity. On the contrary, at FN, BMD was higher than in the reference population, both in eugonadal (1.01 +/- 0.22, P < 0.001) and hypogonadal (0.63 +/- 0.17, P < 0.001) patients only in subjects with active disease, but not in those in which the disease was controlled (eugonadal: 0.31 +/- 0.23, P = ns; hypogonadal 0.04 +/- 0.28, P = ns). We did not observe any difference in BMD values according to gender both at LS (males vs. females -0.02 +/- 0.30 vs. 0.01 +/- 0.24, P = ns) or at FN (0.77 +/- 0.19 vs. 0.63 +/- 0.15, P = ns). CONCLUSIONS: The anabolic effect of GH excess on bone in acromegalic patients is: (i) gender-independent; (ii) evident at the spine only in eugonadal regardless of disease activity; (iii) evident at femoral neck only in the presence of active disease regardless of gonadal status. PMID- 12780750 TI - CTLA4 gene and Graves' disease: association of Graves' disease with the CTLA4 exon 1 and intron 1 polymorphisms, but not with the promoter polymorphism. AB - OBJECTIVE: Recent studies have shown that Graves' disease (GD) is linked to and associated with alleles of the cytotoxic T lymphocyte antigen-4 (CTLA4) locus. However, the true pathogenic polymorphism(s) at this locus remains uncertain. Moreover, the association studies of the promoter CTLA4(-318)C/T polymorphism in white GD populations have produced conflicting results. Therefore, we have analysed three CTLA4 single nucleotide polymorphisms, including promoter CTLA4( 318)C/T, exon 1 CTLA4(49)A/G and intron 1 CTLA4(1822)C/T in our GD cohort from the UK. PATIENTS AND METHODS: We studied 301 white patients with GD and 349 healthy ethnically matched local controls. Amongst GD probands, 129 had significant thyroid-associated orbitopathy (TAO; NOSPECS class III or worse). The CTLA4(-318)C/T, CTLA4(49)A/G and CTLA4(1822)C/T polymorphisms were genotyped by using the restriction enzymes MseI, Bst71I and HaeIII, respectively. RESULTS: We found no association between GD and alleles of CTLA4(-318)C/T. GD was found to be associated with the G allele of CTLA4(49)A/G[P = 5.9 x 10(-6), odds ratio (OR) 1.65] and the T allele of CTLA4(1822)C/T (P = 7.7 x 10(-6), OR 1.64). The frequencies of these alleles were significantly higher in GD probands with significant TAO than in those without TAO (G allele: P = 0.001, OR 1.68; T allele: P = 0.001, OR 1.70). CONCLUSIONS: The promoter CTLA4(-318)C/T polymorphism is not in linkage disequilibrium with the pathogenic polymorphism(s) at the CTLA4 locus. The alleles of both the exon 1 CTLA4(49)A/G and the intron 1 CTLA4(1822)C/T polymorphisms are associated with GD, which is stronger in patients with TAO. PMID- 12780752 TI - Hypothalamic growth hormone-releasing hormone (GHRH) cell number is increased in human illness, but is not reduced in Prader-Willi syndrome or obesity. AB - BACKGROUND: Acute illness leads to increased GH, but reduced IGF-I secretion, while both are reduced in chronic illness. Prader-Willi syndrome (PWS) is a genetic obesity syndrome, with GH deficiency a feature independent of obesity. Reduced GH secretion may result from decreased hypothalamic release of GH releasing hormone (GHRH). OBJECTIVE: To quantify hypothalamic GHRH neurone cell number in control subjects with various lengths of premorbid illness duration, PWS and non-PWS obese subjects. DESIGN: We examined GHRH neurones in the infundibular nucleus/median eminence complex of control subjects (n = 26, including four children), PWS (n = 6) and non-PWS (n = 4) obese adults and PWS children (n = 2), by quantitative immunocytochemistry, using postmortem material. RESULTS: We found: (i) higher GHRH cell number during prolonged illness prior to death in both control adults (r = +0.62, P = 0.002, cell number vs. premorbid illness duration) and PWS adults (r = +0.90, P = 0.02); (ii) higher GHRH cell number in female than male adults [by 53% (95% confidence interval 28-83%) in controls, P = 0.005, correcting for premorbid illness duration]; (iii) no difference in GHRH cell number between PWS adults and control or non-PWS obese adults (P = 0.7 and P = 0.4, adjusting for sex and illness duration); and (iv) low GHRH cell number in only one PWS child (who had been receiving exogenous GH therapy). CONCLUSIONS: These findings suggest continued activation of GHRH neurones during prolonged illness. There is no evidence that the GH deficiency in PWS results from reduced GHRH cell number, and GHRH neuronal responses to illness and exogenous GH treatment appear normal in PWS. PMID- 12780751 TI - Perturbations in adiponectin, leptin and resistin levels in acromegaly: lack of correlation with insulin resistance. AB - BACKGROUND: Insulin resistance, impaired glucose tolerance and type 2 diabetes are common in acromegalic subjects. The mechanism underlying this insulin resistance is unclear. DESIGN: We investigated the levels of the adipocytokines, resistin, adiponectin and leptin in a group of 18 acromegalic subjects and 18 control subjects matched for age, gender and body mass index. RESULTS: Here we demonstrate for the first time significant elevation in adiponectin levels in acromegalic subjects compared to control subjects 12.5 +/- 1.2 vs. 8.97 +/- 1.1 mg/l, P = 0.029. The resistin levels were similar in acromegalic subjects and controls; 20.65 +/- 2.99 vs. 19.03 +/- 4.72 micro g/l. No evidence of a correlation between adiponectin and insulin resistance as calculated from HOMA-R was found. No correlation was observed either between adiponectin or resistin levels and GH levels, total IGF-I or free IGF-I levels. Leptin levels were significantly reduced in acromegalic subjects, 8.22 +/- 2.26 vs. 18.3 +/- 4.1 micro g/l, P = 0.004. In control subjects, significant correlations between leptin levels and HOMA-R and between resistin levels and HOMA-R were observed. These relationships were not apparent in acromegalic subjects. CONCLUSION: From these data we conclude that changes in resistin and adiponectin levels are unlikely to account for the insulin resistance of acromegaly. PMID- 12780753 TI - Hereditary corticosteroid-binding globulin deficiency due to a missense mutation (Asp367Asn, CBG Lyon) in a Brazilian kindred. AB - OBJECTIVE: Abnormal corticosteroid-binding globulin (CBG) is an extremely rare condition and only three mutations have been described in four families. The molecular basis of an abnormal CBG in a Brazilian family was studied and correlations between genotype and serum cortisol, cortisol binding capacity (CBC) and CBG levels were determined. SUBJECTS: All 10 family members, comprising three generations, and nine healthy volunteers were studied. MEASUREMENTS: Genomic DNA was extracted from white blood cells from all family members. The human cbg exons 2-5 were amplified by PCR, submitted to automatic sequencing. Cortisol and CBG levels in serum were measured by radioimmunoassay (RIA). CBC in serum was determined using tritiated cortisol and other cortisol binding parameters were calculated through Scatchard analysis. RESULTS: A missense mutation in exon 5 of cbg (1254G --> A; Asp367Asn), recently described as CBG Lyon, was found in all family members. The proband and one sister were homozygous whereas all other family members, including parents, were heterozygous for this mutation. Cortisol levels in the only two homozygotes were lower than in heterozygotes and both were significantly lower as compared to controls (69 and 182 nmol/l vs. 267 +/- 129 nmol/l vs. 459 +/- 195 nmol/l, respectively, P < 0.05). CBC was decreased in the two homozygotes as compared to heterozygotes and in both groups as compared to controls (< 90 and 114 nmol/l vs. 305.0 +/- 81.4 nmol/l vs. 594.8 +/- 59.5 nmol/l, respectively, P < 0.05). CBG levels were lower in homozygotes as compared to heterozygotes and in both as compared to controls (325 and 375 nmol/l vs. 496.75 +/- 50.75 nmol/l vs. 647.25 +/- 87.50 nmol/l, respectively, P < 0.05). CONCLUSIONS: An abnormal CBG resulting from a missense mutation and known as CBG Lyon was found in this Brazilian kindred. This abnormal CBG has decreased affinity for cortisol and results in low or low normal serum cortisol levels in homozygous and heterozygous subjects. Although relative hypotension and fatigue have recently been associated with CBG deficiency in a family with two CBG mutations (null and Lyon), the two homozygous subjects in this kindred were both normotensive and only the proband presented with fatigue. PMID- 12780754 TI - Postoperative surveillance of clinically nonfunctioning pituitary macroadenomas: markers of tumour quiescence and regrowth. AB - BACKGROUND: Postoperative management of clinically nonfunctioning pituitary adenomas (NFPA) presents difficult challenges. There are no good serum markers for presence or growth of the tumour, medical treatment is not effective and radiotherapy carries the risk of significant side-effects. OBJECTIVE: The purpose of this study was to investigate the natural history and biological behaviour of surgically treated NFPA, with a special effort to identify characteristics indicative of a more aggressive course that could assist in the clinical decision making process. STUDY DESIGN: Patients operated on at our institution for NFPA undergo uniform routine clinical follow-up at the endocrine clinic. Magnetic resonance imaging (MRI) studies are performed 3, 6 and 12 months after transsphenoidal surgery and yearly thereafter for the first 5 years. Subsequently, imaging is performed once every 2 years or as clinically indicated. From 1992 onwards, no patient received immediate postoperative radiation therapy. PATIENTS: One hundred and twenty-two patients (78M/45F) operated on at our institution since 1989 and with a minimal follow-up of 1 year comprised the study group. MEASUREMENTS: Tumour size and characteristics were determined by MRI using a modification of Hardy's and Wilson's classifications. Maximal tumour height was also recorded and the information was routinely stored in a computerized database. RESULTS: Mean (+/- SD) follow-up was 51 +/- 31 months. Fourteen patients received postoperative radiation therapy. Subsequent tumour growth was observed in five of them, reduction in tumour size in four and no size changes in five. One hundred and eight patients did not receive postoperative radiation. Tumour enlargement occurred in 41 of 78 and in six of 30 patients with and without residual tumour after operation (P = 0.0024). The presence of cavernous sinus invasion before surgery [P = 0.02, odds ratio (OR) 2.72; confidence interval (CI) 1.1-6.43] and the extent of suprasellar extension in the postoperative tumour remnant (P = 0.0054 for presence of stage A, OR 4.4; 95% CI 1.5-12.5; and P = 0.012 for presence of stages B or C, OR 16.2; CI 1.8-144) were strong independent predictors of tumour enlargement. CONCLUSION: Our data may ease the selection of patients in whom radiation therapy is likely to be necessary for tumour control, and confirms that close postoperative follow-up is an adequate primary approach in low-risk patients. PMID- 12780755 TI - Short-term effects of prednisolone and dexamethasone on circulating concentrations of leptin and sex hormone-binding globulin in children being treated for acute lymphoblastic leukaemia. AB - OBJECTIVE: Disturbances in body weight regulation are often encountered during glucocorticoid treatment and are associated with increased insulin resistance and truncal fat accumulation. Children were investigated who were receiving glucocorticoid treatment for acute lymphoblastic leukaemia (ALL). They were randomized to receive either prednisolone or dexamethasone as part of induction of remission. This randomization process provided a suitable opportunity to compare the effects of these two administered steroid on surrogate markers of adipocyte activity (leptin) and hyperinsulinaemia/insulin resistance (SHBG). DESIGN AND PATIENTS: Prospective study over 16 weeks of children randomized to receive prednisolone (40 mg/m2) or dexamethasone (6.5 mg/m2) as part of the MRC ALL97/99 induction chemotherapy for ALL. Nineteen children (8 male, 11 female) with a median age 5.9 years (range 2.6-13 years) were recruited into the study. Main outcome measures were body mass index (BMI), serum leptin and sex hormone binding globulin (SHBG). RESULTS: Glucocorticoid administration for 5 weeks resulted in significant (P < 0.05) increases in BMI, leptin (corrected for BMI) and the leptin : SHBG ratio and lowering of SHBG. Dose for dose, dexamethasone was significantly more potent than prednisolone in altering these parameters. CONCLUSIONS: Short-term glucocorticoid treatment has significant effects on BMI, leptin and SHBG. The leptin : SHBG ratio increase indicates that this may be a novel and sensitive biochemical marker of metabolic change. Our results suggest that glucocorticoid treatment regimens should be kept as short as possible to avoid possible detrimental effects associated with increased adiposity and insulin resistance. PMID- 12780756 TI - The circulating IGF system and its relationship with 24-h glucose regulation and insulin sensitivity in healthy subjects. AB - OBJECTIVE AND DESIGN: It has been suggested that circulating free IGF-I participates in glucose homeostasis and that IGFBP-1 reflects changes in insulin sensitivity. To study this further, we examined 10 healthy, nonobese subjects under standardized conditions for 24 h with and without an intravenous infusion of glucose, the latter in order to augment insulin sensitivity. Serum was collected every 2 h for analysis of free and total IGFs, IGFBP-1, - 2 and - 3 and the acid labile subunit (ALS). Insulin sensitivity was estimated at the end of each 24-h study period by use of the hyperinsulinaemic euglycaemic clamp technique. RESULTS: Glucose infusion resulted in mild hyperglycaemia (P < 0.0001), a reduction in IGFBP-1 by approximately 40% (P < 0.0003), and increased insulin and C-peptide levels (P < 0.0001). Glucose infusion also increased insulin sensitivity (P < 0.003). However, despite the reduction in IGFBP-1, glucose infusion did not increase free IGF-I over the control level, and free IGF II was slightly reduced (P < 0.02). Irrespective of glucose infusion, free IGF-I and -II remained stable during daytime (i.e. they were unresponsive to meal related changes in plasma glucose), but both free fractions decreased during the night, reaching nadir at 04.00 h. None of the other members of the IGF system showed any relationship with plasma glucose levels. Finally, we failed to observe any relationship between changes in insulin sensitivity and the circulating IGF system. CONCLUSION: We found no evidence that the circulating IGF system is involved in meal-related blood glucose regulation or that it reflects short-term changes in insulin sensitivity in healthy, nonobese subjects. However, we cannot preclude that the observed changes in circulating IGFBP-1 may affect the glucose lowering effect of IGF-I and -II at the local tissue level. PMID- 12780757 TI - Combined pituitary hormone deficiency in Australian children: clinical and genetic correlates. AB - OBJECTIVE: Mutations in the gene for the POU domain transcription factor POU1F1 (human Pit-1) have been reported in patients with GH, TSH and PRL deficiencies. PROP1 (Prophet of Pit-1) gene mutations also cause gonadotrophin deficiencies and in some cases partial ACTH deficiency. This study analyses the POU1F1 and PROP1 genes in a cohort of Australian children with combined pituitary hormone deficiency (CPHD) and correlates results with patient phenotype. PATIENTS AND DESIGN: Genomic analysis was carried out on 33 patients with CPHD referred from centres around Australia. Clinical data were collected from medical records and referring physicans. RESULTS: POU1F1 mutations were identified in two of four patients with a suggestive phenotype. In a female patient, novel compound heterozygous POU1F1 mutations were identified: Arg143Leu in exon 3 and Leu194Gln in exon 4. This patient presented with failure to thrive at 6 weeks of age and has deficiencies of TSH and GH. A previously described heterozygous Arg271Trp mutation in exon 6 of the POU1F1 gene was identified in a female infant who presented with growth failure and was diagnosed with TSH then GH deficiencies. No PROP1 mutations were identified; however, we describe a number of previously unreported PROP1 polymorphisms. No patients presenting with deficiencies of all anterior pituitary hormones early in life had POU1F1 or PROP1 gene mutations. CONCLUSIONS: In 33 Australian children with CPHD we have identified POU1F1 mutations in two patients and no PROP1 mutations. We speculate that in the majority of children other genes must be responsible for the CPHD phenotype. PMID- 12780758 TI - Cushing's syndrome secondary to topical corticosteroids abuse. PMID- 12780759 TI - Hypophosphatemia, osteomalacia and proximal muscle weakness treated by surgery. PMID- 12780760 TI - North Staffordshire Headache Survey: development, reliability and validity of a questionnaire for use in a general population survey. AB - The North Staffordshire Headache Survey aims to measure the effect and impact of headaches, medicine use and healthcare utilization in a general population sample. A self-reporting questionnaire was piloted in a general population sample, with reliability being tested in a sample of pilot responders after one month and validity by comparing pilot responders with primary and secondary care headache consulters. One hundred and twenty-two (61%) responded to the pilot survey, with 56% of items having completion rates of 90% or more, and tests showed good internal consistency (>90%). One-month test-retest data showed good agreement, though questions relating to specific time periods (with partial or no overlap between survey periods) showed expected lower agreement. The headache consulters reported greater frequency, duration and severity of headaches than the population sample suggesting good construct validity. Results from these studies indicate that the questionnaire is a reliable and valid instrument to collect data about headaches in the general population. PMID- 12780761 TI - Caffeine-induced headache in children and adolescents. AB - Caffeine is the most widely used behaviourally active substance. Excessive caffeine consumption, mostly in the form of coffee and tea, is a well-recognized cause of headache or migraine, and withdrawal can cause headache. Nevertheless, caffeine abuse headache is not listed as a separate category in the International Headache Society classification, 1988. We report our experience with children and adolescents with daily or near-daily headache and excessive consumption of caffeine in the form of cola drinks. Over a period of 5 years we have encountered, in a tertiary headache clinic in a general hospital, 36 children and adolescents (17 girls and 19 boys) with daily or near-daily headache related to excessive caffeine intake in the form of cola drinks. The mean age of the subjects was 9.2 years (range 6-18) and mean headache duration was 1.8 years (range 0.6-5). All were heavy cola drinks consumers; at least 1.5 L of cola drinks per day (192.88 mg of caffeine daily), and an average of 11 (range 10.5 21) L of cola drinks a week, which amounts to 1414.5 mg of caffeine (range 1350.1 2700.3). Patients were encouraged to achieve gradual withdrawal from cola drinks, which led to complete cessation of all headaches in 33 subjects, whereas one boy and two adolescent girls continued to suffer from migraine without aura not frequent enough to justify prophylactic medication. Children and adolescents with high daily caffeine consumption in the form of cola drinks may suffer from caffeine-induced daily headache. Gradual withdrawal can be achieved without withdrawal headache and with complete disappearance of the induced chronic daily headache. PMID- 12780762 TI - Migraine and tension-type headache in Croatia: a population-based survey of precipitating factors. AB - The careful monitoring of the trigger factors of headache could be an important step in treatment, because their avoidance may lessen the frequency and severity of attacks. Furthermore, they may provide a clue to the aetiology of headache. The aim of the present study was to estimate the prevalence of tension-type headache (TTH) and to establish the frequency of precipitating factors in subjects with migraine and TTH in the adult population of Bakar, County of the Coast and Gorski Kotar, Croatia. Another important purpose of the study was to examine the relationship of the precipitating factors with migraine and TTH, and with migraine subtypes: migraine with aura (MA) and migraine without aura (MO). We performed a population-based survey using a 'face-to-face door-to-door' interview method. The surveyed population consisted of 5173 residents aged between 15 and 65 years. The 3794 participants (73.3%) were screened for headache history according to the International Headache Society (IHS) criteria. Headache screen-positive responders, 2475 (65.2%), were interviewed by trained medical students with a structured detailed interview focused on the precipitating factors. The following precipitating factors in lifetime migraineurs and tension type headachers have been assessed: stress, sleep disturbances, eating habits, menstrual cycle, oral contraceptives, food items, afferent stimulation, changes in weather conditions and temperature, frequent travelling and physical activity. A total of 720 lifetime migraineurs and 1319 tension-type headachers have been identified. The most common precipitants for both migraine and TTH were stress and frequent travelling. Stress (odds ratio (OR) 1.4, 95% confidence interval (CI) 1.17, 1.69) was associated with migraine, whereas physical activity (OR 0.72, 95% CI 0.59, 0.87) was related to TTH. Considering MA and MO, frequent travelling (OR 2.2, 95% CI 1.59, 2.99), food items (OR 2.2, 95% CI 1.35, 3.51) and changes in weather conditions and temperature (OR 1.75, 95% CI 1.27, 2.41) exhibited a significant positive association with MA. The present study demonstrated that precipitant-dependent attacks are frequent among both migraineurs and tension-type headachers. Lifetime migraineurs experienced headache attacks preceded by triggering factors more frequently than tension-type headachers. MA was more frequently associated with precipitating factors than MO. We suggest that some triggering factors may contribute to the higher occurrence of precipitant-dependent headache attacks in susceptible individuals. PMID- 12780763 TI - Effect of active treatment is lower when using placebo control in clinical trials on acute therapy of migraine. AB - Methodological aspects of study design affect therapeutic outcome. We tested the hypothesis that treatment effect of an active drug in studies on acute therapy of migraine is lower in placebo-controlled studies than in those not using a placebo controlled design. From 522 eligible studies on acute therapy of migraine cited in Pubmed database which were published between 1964 and 1997 we randomly selected 100 studies for evaluation. We excluded five studies because they did not include a quantitative measurement of pain intensity. Of the 95 studies included in the analysis, 61 used placebo control. Response to active drug was significantly lower (P < 0.05) in placebo-controlled studies (61% [lower and upper quartiles 44% and 75%] vs. 71% [56% and 75%]). We did not find any significant effect of quality of study design on net effect of verum. This observation should be taken into account for future planning of controlled clinical studies in acute migraine. PMID- 12780764 TI - Chronic daily headaches: clinical profile in Indian patients. AB - Chronic daily headache (CDH) remains a relatively unexplored entity in India. Misconceptions are common, unnecessary investigations often done and inappropriate therapy prescribed. Analgesic overuse is seldom recognized. The present report appears to be the first of its kind from India. CDH has been defined as headaches occurring more than 15 days per month for more than 3 months (secondary causes excluded). Over 2 years (1998-1999) 849 cases (49.6% of all primary headaches) were seen. More than 1 year's follow-up data were available in 205 subjects (M 34; F 171). The distribution of these was as follows: (i), chronic tension-type headache (CTH), 33 (16.1%); (ii), chronic/transformed migraine (TM), 169 (82.4%); (iii), new persistent CDH, 3 (1.5%). There were 169 cases of TM (M : F 1 : 4.7; age 26-58 years). History of past episodic migraine was present in all. Transformation had been gradual (89.4%) or acute (10.6%). Possible factors in transformation included psychological stress (44.4%), analgesic overuse (28.4%), ergot overuse (4.1%). HRT seemed to be implicated in three female subjects. Analgesic overuse was limited between intake of 600 and 2400 mg of aspirin equivalent per day (mean 735 mg). Ergot overuse varied between 1 and 3 mg/day of ergotamine for > or = 3 days/week. With medical therapy approximately 70% TM and 40% CTH patients noted significant improvement. About 80% of these relapsed on therapy withdrawal. CDH in India is not uncommon. Analgesic/ergot overuse needs to be recognized early. The average dose of analgesic implicated in CDH seems much less compared with that reported in the West. PMID- 12780765 TI - Abnormal 5-HT1D receptor function in cluster headache: a neuroendocrine study with sumatriptan. AB - The purpose of this study was to assess the sensitivity of 5-HT(1D) receptors in patients with episodic cluster headache using sumatriptan as a pharmacological probe. The drug, a selective 5-HT(1B/1D) agonist, stimulates the secretion of growth hormone and inhibits the release of prolactin, adrenocorticotropic hormone (ACTH) and cortisol. These effects may be used to explore the function of serotonergic systems in vivo. We administered subcutaneous sumatriptan and placebo to 20 patients with cluster headache (10 in the active phase and 10 in the remission period) and to 12 controls. The sumatriptan-induced increase of growth hormone concentrations was significantly (P < 0.05) blunted in patients with active cluster headache. Prolactin and ACTH responses to the drug were significantly (P < 0.05) reduced in patients with cluster headache, both in the active and in the remission period. Our results suggest that cerebral serotonergic functions mediated by 5-HT(1D) receptors are altered in patients with episodic cluster headache. PMID- 12780766 TI - Nociceptive temporalis inhibitory reflexes evoked by CO2-laser stimulation in tension-type headache. AB - The aim of the study was to evaluate the laser-induced suppression periods of the temporalis muscle in patients with tension-type headache, compared with the pattern of temporalis activity suppression induced by electrical stimulation. Fifteen patients with chronic and 10 with episodic tension-type headaches were selected. Suppression periods were recorded simultaneously from both temporalis muscles using both electrical stimuli and CO2-laser stimuli. A significant reduction in the later electrically induced suppression period was found in both tension-type headache groups. Laser stimulation induced a first suppression period (LSP1) with a latency of about 50 ms in all patients. The features of LSP1 were similar across groups. The LSP1 should correspond to the first suppression period induced by electrical stimulus, which is partly a nociceptive response, whereas the second period seemed negligibly linked with the activation of pain related afferents, though probably their activation may contribute to increase the reflex duration and to emphasize abnormalities in tension-type headache. PMID- 12780767 TI - Selective decrease in serotonin synthesis rate in rat brainstem raphe nuclei following chronic administration of low doses of amitriptyline: an effect compatible with an anti-migraine effect. AB - The effects of chronic, low-dose amitriptyline on serotonin (5-HT) synthesis rate were measured in rat brain using autoradiography and the trapping of alpha-[14C] methyl-L-tryptophan (alpha-[14C]-MTrp). Rats received amitriptyline (2 mg/kg per day) or saline via intraperitoneal osmotic minipumps for 21 days. Amitriptyline had no effect on any physiological parameters measured, or on free or total plasma tryptophan levels. However, amitriptyline exerted selective decreases of 15% and 17% (P < 0.001) in serotonin synthesis rates in the dorsal and median raphe nuclei, respectively. There was no reduction in any of the projection areas studied, including the cerebral cortex, hippocampus, thalamus, hypothalamus or striatum. The data suggest that chronic low doses of amitriptyline can lead to sustained 5-HT re-uptake inhibition selectively in the raphe nuclei, an effect compatible with tonic activation of 5-HT(1A) autoreceptors and inhibition of 5-HT synthesis. The failure of chronic amitriptyline treatment to affect 5-HT synthesis rate in the projection areas may ensure an adequate regulation of pain pathways implicated in migraine headache, an effect possibly related to amitriptyline anti-migraine efficacy. PMID- 12780769 TI - Epidemiological and clinical characteristics of migraine and tension type headache in 1146 females in Kayseri, Turkey. AB - In a female population of Turkey (1146 adult females), some epidemiological and clinical characteristics of migraine and tension type headache and their subtypes were investigated. The relation of the headache severity to clinical characteristics were inquired. Migraine prevalence was found to be statistically higher in the 35-44 years age group (P < 0.01) and those who were university graduates (P < 0.001), married (P < 0.01) and living in urban areas (P < 0.01). Tension type headache was found to be higher in the 45-64 years age group (P < 0.05). Chronic tension type headache patients were found to be older than episodic type (P < 0.01) and frequently were in the lowest education level (P < 0.05). Presence of impact on daily activities because of the severity of headache was found to be related to aggravation by physical activities (P = 0.001) in tension type headache, with no clinical characteristics in migraine headache and on consideration of all headache patients with throbbing nature (P < 0.05), aggravation on physical activities (P = 0.001), nausea (P < 0.01), vomiting (P < 0.05) and phonophobia (P < 0.05). PMID- 12780768 TI - A note on cluster headache in a population-based twin register. AB - Evidence of a familial risk factor in cluster headache is accumulating and studies of twin concordance may resolve family resemblance into genetic and environmental influences. The past literature on cluster headache in twins comprises a few case reports of concordant monozygotic pairs. Swedish twin pairs with a diagnosis of cluster headache were selected through a cross-match of national registers of twin births and hospitalizations. Seventeen discordant twin pairs were found, in which it was possible to verify cluster headache status in 11 complete pairs (two monozygotic, four dizygotic, and five unlike-sexed pairs). In both members of a female monozygotic pair, migraine without aura developed after birth of the first child and remitted by menopause, whereas post-menopausal development of chronic cluster headache occurred in only one of them. The importance of individual specific factors for cluster headache was demonstrated. However, to explain familial aggregation a larger sample of affected twin pairs is necessary. PMID- 12780770 TI - Migrainous cerebral infarction in the Sagrat Cor Hospital of Barcelona stroke registry. AB - Nine of 2000 consecutive stroke patients included in the Sagrat Cor Hospital of Barcelona Stroke Registry over a 10-year period fulfilled the strictly defined International Headache Society criteria for migrainous stroke and in whom other causes of stroke were ruled out. They accounted for 13% of all first-ever ischaemic stroke of unusual cause. Migrainous stroke was more common in women (67%) and in patients aged 45 years or younger (78%) compared to the remaining ischaemic strokes of unusual cause. No patient died during hospital stay and 67% were symptom-free at discharge. In the multivariate analysis, nausea or vomiting (odds ratio (OR) 8.40, 95% confidence interval (CI) 1.49-47.21) and age (OR 0.95, 95% CI 0.91-0.99) were predictors of migrainous stroke. Migrainous stroke is a rare entity. Vascular risk factors are uncommon and the prognosis is generally good. Patients with migrainous stroke present some different clinical features from other ischaemic strokes of unusual aetiology. PMID- 12780771 TI - Shortlasting, unilateral, neuralgiform headache attacks with conjunctival injection and tearing (SUNCT syndrome) and tumour of the cavernous sinus. PMID- 12780772 TI - SUNCT syndrome: the first German case series. PMID- 12780773 TI - A patient with cluster headache responsive to indomethacin: any relationship with chronic paroxysmal hemicrania? PMID- 12780774 TI - Adhesion of enteropathogenic Escherichia coli to host cells. AB - Enteropathogenic Escherichia coli (EPEC) adhere to the intestinal mucosa and to tissue culture cells in a distinctive fashion, destroying microvilli, altering the cytoskeleton and attaching intimately to the host cell membrane in a manner termed the attaching and effacing effect. Typical EPEC strains also form three dimensional microcolonies in a pattern termed localized adherence. Attaching and effacing, and in particular intimate attachment requires an outer membrane adhesin called intimin, which binds to the translocated intimin receptor, Tir. Tir is produced by the bacteria and delivered to the host cell via a type III secretion system. In addition to this well-established adhesin-receptor pair, numerous other adhesin interactions between EPEC and host cells have been described including those between intimin and cellular receptors and those involving a bundle-forming pilus and flagella and unknown receptors. Much additional work is needed before a full understanding of EPEC adhesion to host cells comes to light. PMID- 12780775 TI - Show me the substrates: modulation of host cell function by type IV secretion systems. AB - Evidence for the involvement of type IV protein secretion systems in bacterial virulence is accumulating. Many of the substrate proteins secreted by type IV systems either hijack or interfere with specific host cell pathways. These substrates can be injected directly into host cells via the type IV apparatus or are secreted by the type IV machinery in a state that allows them to gain access to cellular targets without the further assistance of the type IV system. Arguably, the protein substrates of most type IV secretion systems remain undiscovered. Here, we review the activities of known type IV substrates and discuss the putative roles of unidentified substrates. PMID- 12780777 TI - Structural definition on the surface of Helicobacter pylori type IV secretion apparatus. AB - Genetic and functional studies have indicated that the type IV secretion system (TFSS) of Helicobacter pylori forms a secretion complex in the cell envelope that protrudes towards the outside in order to inject CagA protein into gastric epithelial cells. However, the proposed structural model is based on partial amino acid homology with the components of the Agrobacterium tumefaciens TFSS. Therefore, we undertook the identification of the structural features of the TFSS exposed on the surface of H. pylori and found that filamentous structures present on the bacterial surface are related to the secretion apparatus. Using immunofluorescence microscopy with antibodies directed to tyrosine-phosphorylated CagA (pY-CagA) and Hp0532 (VirB7) in the infection assay, pY-CagA signals were detected just below the host cell-attached bacteria, where Hp0532 (VirB7) signals were detected as co-localized, suggesting that the CagA injected into the host cell through the TFSS apparatus is still mostly confined to the areas just below the attached bacteria after being phosphorylated. Furthermore, the filamentous structures on bacterium were found to be associated with Hp0532 (VirB7) or Hp0528 (VirB9), the major components of TFSS, by immunogold electron microscopy. These results strongly suggest that the H. pylori TFSS apparatus is a filamentous macromolecular structure protruding from the bacterial envelope. PMID- 12780776 TI - Actin-based motility of Burkholderia pseudomallei involves the Arp 2/3 complex, but not N-WASP and Ena/VASP proteins. AB - The facultative intracellular bacterium Burkholderia pseudomallei induces actin rearrangement within infected host cells leading to formation of actin tails and membrane protrusions. To investigate the underlying mechanism we analysed the contribution of cytoskeletal proteins to B. pseudomallei-induced actin tail assembly. By using green fluorescent protein (GFP)-fusion constructs, the recruitment of the Arp2/3 complex, vasodilator-stimulated phosphoprotein (VASP), Neural Wiskott-Aldrich syndrome protein (N-WASP), zyxin, vinculin, paxillin and alpha-actinin to the surface of B. pseudomallei and into corresponding actin tails was studied. In addition, antibodies against the same panel of proteins were used for immunolocalization. Whereas the Arp2/3 complex and alpha-actinin were incorporated into B. pseudomallei-induced actin tails, none of the other proteins were detected in these structures. The overexpression of an Arp2/3 binding fragment of the Scar1 protein, shown previously to block actin-based motility of Listeria, had no effect on B. pseudomallei tail formation. Infections of either N-WASP- or Ena/VASP-defective cells showed that these proteins are not essential for B. pseudomallei-induced actin polymerization. In conclusion, our results suggest that B. pseudomallei induces actin polymerization through a mechanism that differs from those evolved by Listeria, Shigella, Rickettsia or vaccinia virus. PMID- 12780778 TI - Deficiency in mycolipenate- and mycosanoate-derived acyltrehaloses enhances early interactions of Mycobacterium tuberculosis with host cells. AB - Lipids that are uniquely found in the cell envelope of pathogenic mycobacteria, such as those containing multiple methyl-branched long-chain fatty acids, have long been thought to play a role in host-pathogen interactions. The recent construction by Dubey et al. (2002) Mol Microbiol 45: 1451-1459, of a Mycobacterium tuberculosis mutant that is deficient in the synthesis of the di- and tri-methylbranched fatty acids, mycolipenates and mycosanoates, found in some forms of diacyltrehaloses (DAT) and polyacyltrehaloses (PAT) provided the opportunity to assess the contribution of these complex lipids to pathogenesis directly. We provide evidence that DAT/PAT deficiency affects the surface global composition of the mycobacterial cell envelope improving the efficiency with which M. tuberculosis binds to and enters phagocytic and non-phagocytic host cells. Interestingly, this property did not affect the overall replication and persistence of the tubercle bacillus in the lungs, spleen and liver of mice infected via the respiratory or intravenous route. PMID- 12780780 TI - Amyloid deposits in the bone marrow. PMID- 12780779 TI - Src kinase has a central role in in vitro cellular internalization of Staphylococcus aureus. AB - Traditionally recognized as an extracellular pathogen, the Gram-positive bacterium Staphylococcus aureus can also be internalized by a variety of cell types in vitro. Internalization is known to involve binding of the host extracellular protein fibronectin to the bacterium, recognition of the fibronectin-coated bacterium by the fibronectin-binding integrin alpha5beta1 on the host cell surface, and integrin-mediated internalization. Here we examine elements of mammalian cell signalling pathways involved in S. aureus internalization. The mouse fibroblast cell line GD25, in which the gene encoding the beta1 integrin subunit is inactivated, has been complemented with a beta1 integrin cDNA encoding a tyrosine (Y) to phenylalanine (F) mutation in each of the two beta1 integrin intracellular NPXY motifs. This cell line, GD25beta1 A Y783/795F, is defective in migration on fibronectin coated surfaces and intracellular signalling activities involving the tyrosine kinase Src. GD25beta1 A Y783/795F cells have a decreased ability to internalize S. aureus compared to GD25beta1 A cells expressing wild-type beta1 integrins. Furthermore, using mouse embryo fibroblasts in which different members of the Src family kinases are genetically inactivated, we demonstrate that optimal internalization is dependent on expression of Src kinase. Interferon, which has been implicated in repression of the effects of the viral homologue of Src inhibits internalization of S. aureus indicating that internalization may be blocked by inhibitors of Src kinase function. We then demonstrate that Src family kinase specific inhibitors effectively block S. aureus internalization into HeLa cells leading to the conclusion that a function unique to Src is required for optimal internalization of S. aureus in vitro. PMID- 12780781 TI - Plasmacytoma occurring in scar tissue. PMID- 12780782 TI - Angioimmunoblastic T-cell lymphoma. PMID- 12780783 TI - Mechanism of action of purine analogues in chronic lymphocytic leukaemia. PMID- 12780784 TI - Six characters in search of an author: the history of the nomenclature of coagulation factors. PMID- 12780785 TI - Cytotoxic drugs enhance the ex vivo sensitivity of malignant cells from a subset of acute myeloid leukaemia patients to apoptosis induction by tumour necrosis factor receptor-related apoptosis-inducing ligand. AB - We have studied the actions of tumour-necrosis-factor-related apoptosis-inducing ligand (TRAIL) on cells isolated from patients with acute myeloid leukaemia (AML). Apoptosis induction was initially assessed by quantitative morphological analysis. Only 2/19 isolates showed a > 10% increase in apoptotic cells following TRAIL treatment. However, incubation with TRAIL combined with fludarabine, cytosine arabinoside or daunorubicin resulted in additive or super-additive apoptosis induction in approximately half of the isolates. Molecular evidence of super-additive apoptosis induction by TRAIL and cytotoxic agents was obtained by quantification of caspase 3 activation, detected by Western blot analysis of poly (ADP ribose) polymerase cleavage. The ability of TRAIL and daunorubicin to induce super-additive apoptosis correlated with the ability of these agents to activate caspase 8 and to augment cellular levels of the truncated pro-apoptotic form of the BCL-2 family member BID. Our data suggest that co-administration of TRAIL with conventional cytotoxic drugs may be of therapeutic value in some patients with AML. PMID- 12780786 TI - HLA-DRB1*16-restricted recognition of myeloid cells, including CD34+ CML progenitor cells. AB - The therapeutic effect of a human leucocyte antigen (HLA)-identical allogeneic stem cell transplantation (allo-SCT) for the treatment of haematological malignancies is mediated partly by the allogeneic T cells that are administered together with the stem cell graft. Chronic myeloid leukaemia (CML) is particularly sensitive to this graft-versus-leukaemia (GVL) effect. Several studies have shown that in allogeneic responses both CD4 and CD8 cells are capable of strong antigen-specific growth inhibition of leukaemic progenitor cells, but that CD4 cells mainly exert the GVL effect against CML. Efficient activation of allogeneic CD4 cells, as well as CD8 cells, may explain the sensitivity of CML cells to elimination by allogeneic T cells. Identification of the antigens recognized by CD4 cells is crucial in understanding the mechanism through which CML cells are so successful in activating allogeneic T cells. In the present report, we describe the characterization of an allogeneic CD4 T-cell clone, DDII.4.4. This clone was found to react against an antigen that is specifically expressed in myeloid cells, including CD34+ CML cells. The antigen recognition is restricted by HLA-DRB1*16. To our knowledge, this is only the second report on an allogeneic CD4 T-cell clone that reacts with early CD34+ myeloid progenitor cells. PMID- 12780787 TI - P-glycoprotein and BCL-2 levels predict outcome in adult acute lymphoblastic leukaemia. AB - Concurrent resistance mechanisms, such as P-glycoprotein (PGP) and bcl-2, may contribute to a worse outcome in adult acute lymphoblastic leukaemia (ALL). Between 1990 and 2000, we analysed PGP and bcl-2 by flow cytometry, using two anti-PGP (C219 and JSB-1) monoclonal antibodies (mAbs) and an anti-bcl-2 mAb in 115 de novo adult ALL patients. Both a longer overall survival (OS) and longer disease-free survival (DFS) were observed in PGP-negative patients (23%vs 0% at 3 years, P = 0.011 and 29%vs 0% at 2 years, P = 0.006 for C219 respectively; 42%vs 0% at 1.5 years, P = 0.004 and 53%vs 0% at 8.5 months, P = 0.00006 for JSB-1 respectively). Bcl-2 positivity was associated with a significantly higher complete remission rate (90%vs 66%, P = 0.01). Moreover, in 69 patients not presenting with either t(9;22) or B-mature immunophenotype, PGP negativity (JSB 1) maintained its significant favourable prognostic impact with regard to OS (41%vs 0% at 1.5 years, P = 0.009) and DFS (83%vs 0% at 6 months, P = 0.0005). Importantly, within a subset of 62 patients with normal (n = 31) or unknown (n = 31) karyotype, PGP (JSB-1)-negative patients showed both a significantly longer OS and DFS (63%vs 0% at 1.4 years, P = 0.018 and 84%vs 0% at 6 months, P = 0.001 respectively). In multivariate analysis, JSB-1 (P = 0.008) and cytogenetics (P = 0.02) were found to be independent prognostic factors with regard to DFS. Therefore, in adult ALL, PGP and bcl-2 represent sensitive indicators of clinical outcome, and potential targets of novel molecules aimed at overcoming chemoresistance and recurrent relapses. PMID- 12780788 TI - Growth regulation by p27Kip1 is abrogated by multiple mechanisms in aggressive malignant lymphomas. AB - The cyclin-dependent kinase inhibitor p27Kip1 is a key regulator of the G1/S transition, and an inverse relationship between p27Kip1 protein expression and proliferation index has been reported in malignant lymphomas. However, a subset of aggressive B-cell lymphomas demonstrates high p27Kip1 expression despite a high proliferation index. The aim of this study was to determine potential mechanisms by which lymphoma cells abrogate the growth inhibitory effect of high p27Kip1. The effect of transforming growth factor-beta (TGF-beta) and serum stimulation on p27Kip1 expression and cyclin E/cdk2 activity was investigated in four lymphoma cell lines, Jurkat, CEM-6, OCI-Ly1 and Nalm-6. Reactive lymphocytes responded to growth inhibitory TGF-beta by inducing p27Kip1 expression, with subsequent accumulation of cells in G0/G1. In contrast, TGF-beta did not alter the level of p27Kip1 in Jurkat, CEM-6 and OCI-Ly1 cells with no change in cyclin E/cdk2-kinase activity. Serum stimulation also did not result in a significant change in p27Kip1 expression. Western blot analysis of subcellular fractions demonstrated cytoplasmic p27Kip1, corroborated by immunocytochemistry in a subset of the lymphoma cells. Sequestration of p27Kip1 by cyclin D3 was observed in the nuclear and cytoplasmic fractions of Nalm-6, OCI-Ly-1 and NCEB cells. These results indicate that multiple mechanisms contribute to the abrogation of growth regulation by unscheduled high p27Kip1 protein expression including deficient response to TGF-beta and serum, sequestration by cyclin D3 and cytoplasmic displacement. PMID- 12780789 TI - Criteria for the classification of monoclonal gammopathies, multiple myeloma and related disorders: a report of the International Myeloma Working Group. AB - The monoclonal gammopathies are a group of disorders associated with monoclonal proliferation of plasma cells. The characterization of specific entities is an area of difficulty in clinical practice. The International Myeloma Working Group has reviewed the criteria for diagnosis and classification with the aim of producing simple, easily used definitions based on routinely available investigations. In monoclonal gammopathy of undetermined significance (MGUS) or monoclonal gammopathy, unattributed/unassociated (MG[u]), the monoclonal protein is < 30 g/l and the bone marrow clonal cells < 10% with no evidence of multiple myeloma, other B-cell proliferative disorders or amyloidosis. In asymptomatic (smouldering) myeloma the M-protein is >/= 30 g/l and/or bone marrow clonal cells >/= 10% but no related organ or tissue impairment (ROTI)(end-organ damage), which is typically manifested by increased calcium, renal insufficiency, anaemia, or bone lesions (CRAB) attributed to the plasma cell proliferative process. Symptomatic myeloma requires evidence of ROTI. Non-secretory myeloma is characterized by the absence of an M-protein in the serum and urine, bone marrow plasmacytosis and ROTI. Solitary plasmacytoma of bone, extramedullary plasmacytoma and multiple solitary plasmacytomas (+/- recurrent) are also defined as distinct entities. The use of these criteria will facilitate comparison of therapeutic trial data. Evaluation of currently available prognostic factors may allow better definition of prognosis in multiple myeloma. PMID- 12780790 TI - Paediatric myelodysplastic syndromes and juvenile myelomonocytic leukaemia in the UK: a population-based study of incidence and survival. AB - We aimed to identify and classify cases of paediatric myelodysplastic syndromes (MDS) occurring in Britain to estimate the incidence of this rare group of diseases, investigate the results of therapy and identify prognostic risk factors. Patients aged below 15 years at diagnosis were collected from England, Scotland and Wales, inclusively between 1990 and 1999. One hundred and thirty five patients were accepted as de novo MDS or juvenile myelomonocytic leukaemia (JMML). The incidence for this period was 1.35 per million (age standardized rate) which is below that reported outside the UK. The overall survival was 45%[standard error (SE) = 4%] at 5 years: 40% (SE = 6%) for JMML and 50% (SE = 6%) for other MDS. Significant adverse prognostic factors for JMML were a platelet count < 40 x 10(9)/l, raised fetal haemoglobin, FPC score and age above 2 years at diagnosis, for other MDS only monosomy 7 was significant. To conclude, the incidence of MDS/JMML in children in the UK appears to be lower than that reported outside the UK. This may be either a real difference in incidence or variation in reporting. Monosomy 7 is associated with poor outcome in MDS other than JMML. The prognosis of JMML depends on age, platelet count and fetal haemoglobin. PMID- 12780791 TI - Thalidomide and dexamethasone for resistant multiple myeloma. AB - Between November 1998 and April 2000, the combination of thalidomide and dexamethasone was evaluated in 47 consecutive patients with multiple myeloma that was resistant to prior high-dose dexamethasone-based therapies. Remission was observed in 22 patients (47%), including six patients with complete remission. Side-effects were frequent, mild and usually reversible, but deep vein thrombosis occurred in 8% of patients. Survival and remission times were longer among patients treated for previous resistant disease rather than for resistant relapse. This experience supports the use of thalidomide-dexamethasone in myeloma patients with resistant disease and justifies further trials in newly diagnosed patients. PMID- 12780792 TI - Prevalence and prognostic significance of sMUC-1 levels in plasma cell dyscrasias. AB - High serum Mucin-1 (sMUC-1) levels have been shown in patients with adenocarcinoma and multiple myeloma (MM). We evaluated sMUC-1 levels in 76 patients with MM, 6 with plasma cells leukaemia (PCL) and 89 with monoclonal gammopathy of undetermined significance, to establish prevalence data and verify its possible prognostic role. Of the 171 patients, 27 [16%; 95% confidence interval (CI): 10-21%] had high sMUC-1 levels compared with healthy subjects (1.5%; 95% CI: 0-4%). Elevated sMUC-1 levels in MM and PCL patients correlated with anaemia and elevated serum lactate dehydrogenase levels; these patients showed a shorter survival than those with normal sMUC-1 levels (median overall survival: 25 vs. 49 months, P = 0.003). PMID- 12780793 TI - Incidence and prognosis of c-KIT and FLT3 mutations in core binding factor (CBF) acute myeloid leukaemias. AB - DNA from 110 adult de novo acute myeloid leukaemia (AML) patients exhibiting either inv(16) (n = 63) or t(8;21) (n = 47) was screened for mutations in the c KIT (exon 8 and Asp816) and FLT3 (ITD and Asp835) genes. c-KIT exon 8 mutations were found in 15/63 (23.8%) inv(16) patients and 1/47 (2.1%) t(8;21) patients. c KIT Asp816 mutations were present in 5/63 (7.9%) inv(16) AML and 5/47 (10.6%) t(8;21) AML. FLT3 mutations were identified in five patients (7.9%) with inv(16) and three patients (5.6%) with t(8;21) AML. All mutations were mutually exclusive; 40% of inv(16) AML patients possessed either a c-KIT or FLT3 mutation. c-KIT exon 8 mutations were shown to be a significant factor adversely affecting relapse rate. PMID- 12780794 TI - Venous thromboembolism associated with the management of acute thrombotic thrombocytopenic purpura. AB - Venous thromboembolism (VTE) is not a feature of thrombotic thrombocytopenic purpura (TTP), but there has been a recent report of VTE in association with plasma exchange (PEX) treatment for TTP using the solvent detergent (SD) plasma, PLAS+SD. We reviewed the occurrence of VTE in 68 consecutive patients with TTP (25 men, 43 women). Eight documented VTE events [six deep venous thromboses (DVTs), three pulmonary emboli] were identified in seven patients (all female) during PEX therapy. All six DVTs were associated with central lines at the site of thrombosis. Other known precipitating factors included pregnancy, immobility, obesity and factor V Leiden heterozygosity. VTE occurred at a mean of 53 d following the first PEX. The European SD plasma, Octaplas was the last plasma to be used in PEX prior to the VTE in 7/8 events. This is the first report of VTE following Octaplas infusion. VTE is a multifactorial disease and, although several known precipitating factors were present in all patients in this study, the use of large volumes of SD plasma in PEX may be an additional risk factor. We recommend prevention of VTE with graduated elastic compression stockings (class I) at diagnosis and prophylactic low-molecular-weight heparin once the platelet count rises above 50 x 10(9)/l. PMID- 12780795 TI - Biological and clinical features of low-molecular-weight heparin-induced thrombocytopenia. AB - Heparin-induced thrombocytopenia (HIT) is a common adverse effect of unfractionated heparin (UFH) therapy. In contrast, only a few patients have been reported with HIT following low-molecular-weight heparin (LMWH) therapy (LMW HIT). To define the clinical and biological characteristics of LMW-HIT, 180 patients treated for suspected HIT at 15 French centres were investigated. Clinical history was recorded and HIT was confirmed in 59 patients with positive serotonin release assay results: 57 of them had high levels of antibodies (Abs) to heparin-platelet factor 4 complexes (H/PF4) and two had Abs to interleukin 8. Eleven patients were treated exclusively with LMWH (LMW-HIT) and 48 with UFH either alone (UF-HIT, n = 34) or combined with LMWH (UF/LMW-HIT, n = 14). The LMW HIT and UF-HIT groups were similar with respect to sex, age, platelet count before heparin therapy, frequency of bleeding and occurrence of disseminated intravascular coagulation. The interval to onset of HIT was longer in LMW-HIT patients compared with UF-HIT patients (P = 0.03). Severe thrombocytopenia (platelets < 15 x 10(9)/l) was more frequent in the LMW-HIT group (P = 0.04). Thrombosis occurred in three of 11 LMW-HIT patients, i.e. as frequently as in UF HIT patients. LMW-HIT is potentially severe and may be observed after longer heparin treatment compared with UF-HIT. It is highly recommended, therefore, that platelet counts be monitored carefully whenever LMWH is administered. PMID- 12780796 TI - Functional inhibitory human leucocyte antigen class I receptors on natural killer (NK) cells in patients with chronic NK lymphocytosis. AB - Chronic natural killer (NK) lymphocytosis is a rare disorder characterized by an indolent clinical course. Despite high NK cell numbers, many patients present with only mild clinical symptoms, and are often asymptomatic. NK cells are equipped with a range of receptors that bind human leucocyte antigen (HLA)-class I molecules. The killer immunoglobulin-like receptors (KIR, CD158) bind groups of HLA alleles, the CD94/NKG2 receptors bind HLA-E, and the CD85j (ILT2, LIR-1) receptor binds to the relatively non-polymorphic alpha3 domain of HLA molecules. Inhibitory HLA class I receptors silence NK cells against cells expressing normal levels of HLA class I. Analysis of NK cells in six patients with chronic NK lymphocytosis revealed a high level of the inhibitory CD94/NKG2A receptor on all NK cells. In four patients, KIR were absent, in one patient a single KIR was expressed in the absence of self-ligand, and in one patient CD85j and multiple KIR were expressed. Cytotoxicity assays demonstrated that all HLA class I receptors were functional. The ability of monoclonal antibodies to block the receptors and allow killing of autologous target cells established that both receptor and ligand expression were adequate for inhibitory function. We propose that the silent behaviour of NK cells in patients with chronic NK lymphocytosis is due to effective inhibitory HLA class I receptors. PMID- 12780797 TI - Common cytological and cytogenetic features of Epstein-Barr virus (EBV)-positive natural killer (NK) cells and cell lines derived from patients with nasal T/NK cell lymphomas, chronic active EBV infection and hydroa vacciniforme-like eruptions. AB - In this study, we describe the cytological and cytogenetic features of six Epstein-Barr virus (EBV)-infected natural killer (NK) cell clones. Three cell clones, SNK-1, -3 and -6, were derived from patients with nasal T/NK-cell lymphomas; two cell clones, SNK-5 and -10, were isolated from patients with chronic active EBV infection (CAEBV); and the other cell clone, SNK-11, was from a patient with hydroa vacciniforme (HV)-like eruptions. An analysis of the number of EBV-terminal repeats showed that the SNK cell clones had monoclonal EBV genomes identical to the original EBV-infected cells of the respective patients, and SNK cells had the type II latency of EBV infection, suggesting that not only the cell clones isolated from nasal T/NK-cell lymphomas but also those isolated from CAEBV and HV-like eruptions had been transformed by EBV to a certain degree. Cytogenetic analysis detected deletions in chromosome 6q in five out of the six SNK cell clones, while 6q was not deleted in four control cell lines of T-cell lineage. This suggested that a 6q deletion is a characteristic feature of EBV positive NK cells, which proliferated in the diseased individuals. The results showed that EBV-positive NK cells in malignant and non-malignant lymphoproliferative diseases shared common cytological and cytogenetic features. PMID- 12780798 TI - Is macrocytosis a favourable prognostic factor in myelodysplastic syndrome patients without increased blasts? PMID- 12780799 TI - Novel combination chemotherapy for primary (AL) amyloidosis myeloma: clinical, laboratory and serum amyloid-P protein scan improvement. PMID- 12780800 TI - Successful treatment of multicentric Castleman's disease with bilateral orbital tumour using rituximab. PMID- 12780825 TI - Abstracts of the Annual Meeting of the British Association of Urological Surgeons, 23-27 June 2003, Manchester, United Kingdom. PMID- 12780826 TI - Where are the 'poster boys' for bladder cancer? PMID- 12780827 TI - Sexual dysfunction in men with lower urinary tract symptoms and benign prostatic hyperplasia: an emerging link. PMID- 12780828 TI - Lithotripsy for ureteric stones: throw away the ureteroscope. PMID- 12780829 TI - Urinary markers in bladder cancer. PMID- 12780830 TI - Contralateral adrenal metastasis of renal cell carcinoma: treatment, outcome and a review. AB - OBJECTIVE: To report the surgical treatment of patients with renal cell carcinoma (RCC) metastatic to the contralateral adrenal gland and compare our experience with previous reports, as such metastases are found in 2.5% of patients with metastatic RCC at autopsy, and the role of resecting metastatic RCC at this site is not well defined. PATIENTS AND METHODS: We retrospectively identified 11 patients who had surgery for metastatic RCC to the contralateral adrenal gland between October 1978 and April 2001. The patients' medical records were reviewed for clinical, surgical and pathological features, and the patients' outcome. RESULTS: The mean (median, range) age of the patients at primary nephrectomy was 60.9 (64, 43-79) years; all had clear cell (conventional) RCC. Synchronous contralateral adrenal metastasis occurred in two patients. The mean (median, range) time to contralateral adrenal metastasis after primary nephrectomy for the remaining nine patients was 5.2 (6.1, 0.8-9.2) years. All patients were treated with adrenalectomy; there were no perioperative complications or mortality. Seven patients died from RCC at a mean (median, range) of 3.9 (3.7, 0.2-10) years after adrenalectomy for contralateral adrenal metastasis; one died from other causes at 3.4 years, one from an unknown cause at 1.7 years and two were still alive at the last follow-up. CONCLUSIONS: The surgical resection of contralateral adrenal metastasis from RCC is safe; although most patients died from RCC, survival may be prolonged in individual patients. Hence, in the era of cytoreductive surgery, the removal of solitary contralateral adrenal metastasis seems to be indicated. PMID- 12780831 TI - Identifying under-performing surgeons. AB - OBJECTIVE: To estimate the likelihood of poor surgical results being explained by chance rather than under-performance. METHODS: The 30-day mortality rates after radical cystectomy for bladder cancer were analysed theoretically. Surgical competence was defined as a mortality rate of 4%, excellence as 2% and under performance as 8%, 12%, 20% or 40%. Four scenarios were explored for surgeons of different competence: first, the sample size required to show that a given level of under-performance is very unlikely to be due to chance; second, the likelihood of two or more consecutive deaths in a series of cases; third, the likelihood of clustering of deaths, defined as two deaths in five or in 10 cases; and last, the likelihood of outstanding surgical results (i.e. no deaths) being achieved in small cohorts by surgeons of differing competence. RESULTS: For surgeons with a mortality rate of 8%, 12%, 20% or 40%, the sample sizes needed to prove under performance are 211, 65, 21 and seven, respectively. For consecutive deaths, 0.4% of excellent, 1.4% of competent and 21% of surgeons with a mortality rate of 12% will experience two or more consecutive deaths in the next 10 cases. For clustered deaths, 1% of excellent, 5% of competent and 23% of seriously under performing surgeons (mortality rates > or = 12%) will experience two deaths in their next 10 cases. Lastly, for the likelihood of outstanding results, only 3.6% of surgeons with an 8% mortality rate and < 1% of surgeons with a mortality rate > or = 12% will experience no deaths over 40 consecutive cases. CONCLUSIONS: Very large cohorts are needed to confirm even significant under-performance. Consecutive deaths are very unlikely events for competent surgeons. Clustered deaths (two deaths in 10 cases) are very unlikely events for excellent surgeons but plausible for competent ones. Analysis of consecutive/clustered deaths is limited by low statistical sensitivity; only up to a quarter of seriously under performing surgeons are identified. No deaths in 40 consecutive cases implies competence. PMID- 12780832 TI - Problems with prostate specific antigen screening for prostate cancer in the primary healthcare setting in South Africa. AB - OBJECTIVES: To assess the feasibility of detecting early-stage prostate cancer in the primary healthcare setting, and to investigate whether there is a higher incidence of prostate cancer in Black African men. PATIENTS AND METHODS: The study was a collaboration with registrars in the authors' institutions and primary healthcare centres serving mainly a Black African or mixed ancestry (Coloured) population in the semi-urban Cape Town metropolitan area of South Africa. Men aged 50-70 years attending the clinics were counselled about prostate cancer and invited to have a digital rectal examination (DRE), serum prostate specific antigen (PSA) assay and transrectal ultrasonography-guided sextant prostate biopsy if the DRE was clinically suspicious of malignancy or the serum PSA was > or = 4.0 ng/mL. An American Urological Association Symptom Index (AUA SI) was obtained, and urine analysed using dipsticks. RESULTS: From May 2000 to November 2001, 660 men were assessed (mean age 59.4 years, range 30-82); 60.6% were Black African, 37.3% mixed (Coloured), 1.8% White (Caucasian) and 0.2% Asian (Indian). The mean (range) AUA-SI was 5.98 (0-35) in the whole group; the DRE was recorded as clinically suspicious of malignancy in 3.2%. The mean PSA was 20.39 (0.04-10 000) ng/mL in the whole group, but when two outliers (1865 and 10 000 ng/mL) were disregarded, it was 2.4 ng/mL. In Black patients the mean PSA was 31.8 (0.04-10 000) ng/mL, and without the outliers, 2.1 ng/mL; in Coloured patients it was 2.94 (0.05-50) ng/mL. The PSA was > or = 4.0 ng/mL in 9.6% of the whole group, in 7.8% of Black and in 13% of Coloured patients. Prostate biopsies were taken in 21 patients (3.2% of the whole group and a third of those with a PSA of > or = 4.0 ng/mL); in Black patients, biopsies were taken in 1.5% and 19.4%, respectively, and in Coloured patients in 6.1% and 46.9%, respectively. The prostate biopsy showed cancer in 43% of the whole group, in a third of Black and in 47% of Coloured patients; prostate cancer was detected in 1.4%, 0.5% and 2.8%, respectively. CONCLUSIONS: That prostate biopsies were obtained in only 19% of Black and in only 47% of Coloured men with a serum PSA of > or = 4.0 ng/mL is of concern. This indicates that there is a significant problem in getting men with an elevated serum PSA level to undergo a prostate biopsy in the primary healthcare setting in South Africa. PMID- 12780833 TI - A systematic review and meta-analysis of familial prostate cancer risk. AB - OBJECTIVE: To identify published studies quantifying familial prostate cancer risks in relatives of prostate cancer cases and, by meta-analysis, obtain more precise estimates of familial risk according to the family history. METHODS: Thirteen case-control and cohort studies were identified which have reported risks of prostate cancer in relatives of prostate cancer cases. Pooled estimates of risk for various categories of family history were obtained by calculating the weighted average of the log relative risk (RR) estimates from studies. RESULTS: The pooled RR (95% confidence interval) in first-degree relatives was 2.5 (2.2 2.8). There was evidence that this was highest in relatives of cases diagnosed before age 60 years and that RRs declined with age. The risk for the few men with two affected relatives was increased 3.5-fold (2.6-4.8). RRs to sons of cases appeared to be lower than in brothers; a complete explanation of this observation is uncertain. CONCLUSION: Men with a family history of prostate cancer have a significantly greater risk of developing prostate cancer than those with no such history. Risks are greatest for relatives of cases diagnosed when young and those with more than one relative affected. PMID- 12780834 TI - Improvement in urinary symptoms after radical prostatectomy: a prospective evaluation of flow rates and symptom scores. AB - OBJECTIVE: To evaluate the change in flow patterns and urinary symptoms before and after radical retropubic prostatectomy (RRP). PATIENTS AND METHODS: Between 1994 and 1998 one surgeon undertook RRP in 125 men. Urinary flow rates and the International Prostate Symptom Score (IPSS) were recorded before and at each visit after RRP; only voids of > 150 mL were included. Strictures and bladder neck stenoses requiring surgical intervention were noted. Statistical significance was determined using Student's t or the chi-squared test. RESULTS: Before RRP 38% men had a flow rate of < or = 10 mL/s, suggesting obstruction. At the first review (median 2 months) there was an increase in flow rate (median 16.8 vs 11.6 mL/s, P < 0.001) and at the 6-, 14- and 20-month visits this improved further, to 20, 21 and 24 mL/s, respectively. Before RRP 56% of men had moderate or severe symptoms, with an IPSS of > or = 8. At 2, 6, 14 and 20 months the proportion of men with an IPSS of > or = 8 decreased to 26%, 14.5%, 18% and 14% (P < 0.001); 20% developed stricture/stenosis and initially these men had a decrease in flow rate and a higher IPSS. Their symptoms improved when the stricture was treated. CONCLUSIONS: Two-fifths of men with prostate cancer undergoing RRP have bladder outlet obstruction, as defined by a flow rate of < 10 mL/s and bothersome symptoms. This study showed that there is a very significant increase in flow rate and decrease in IPSS after surgery. RRP offers improved voiding function and urinary symptoms, and the possibility of curing the cancer. PMID- 12780835 TI - Morbidity of transrectal ultrasonography-guided prostate biopsies in patients after the continued use of low-dose aspirin. AB - OBJECTIVE: To determine whether low-dose aspirin increases morbidity after transrectal ultrasonography-guided sextant prostate biopsy. PATIENTS AND METHODS: In a single-centre prospective cohort study of 200 patients who underwent sextant prostate biopsies, those routinely taking low-dose aspirin were encouraged to continue to do so before and after biopsy. The morbidity in each case was assessed using a standardized questionnaire that patients completed in the 7 days after biopsy. The presence of haematuria, rectal bleeding and haematospermia were recorded. The questionnaire also directed the patient to record fevers, use of analgesia and any further treatment received. RESULTS: In all, 36 patients took aspirin whilst the other 141 did not. There were no major complications in either group. Of the patients on aspirin, 20 (56%) had haematuria, compared with 83 (59%) of those not taking aspirin (difference 3%, 95% confidence interval, CI, 15 to 21). Overall bleeding (haematuria, rectal bleeding and haematospermia) occurred in 22 patients (61%) of the aspirin group and 105 (74%) of the other group (difference 13%, 95% CI -4 to 31). Comparisons of other morbidities between the groups are also discussed. CONCLUSIONS: There was no statistically significant difference in the incidence of haematuria or overall bleeding after biopsy between the groups. There is no evidence that aspirin needs to be discontinued before sextant prostate biopsy. PMID- 12780836 TI - A 4-year follow-up of a randomized prospective study comparing transurethral electrovaporization of the prostate with neodymium: YAG laser therapy for treating benign prostatic hyperplasia. AB - OBJECTIVE: To compare the safety, efficacy and durability of neodymium (Nd):YAG laser prostatectomy with transurethral electrovaporization of the prostate (TUVP) for treating benign prostatic hyperplasia (BPH). PATIENTS AND METHODS: From March 1995 to March 1997, 180 patients with bladder outlet obstruction secondary to BPH were randomized equally either to Nd:YAG laser therapy or TUVP. Laser therapy combined two different techniques, side-fire coagulation of the lateral lobes and contact vaporization of the median lobe. Before treatment the two groups had a comparable International Prostate Symptom Score (IPSS), quality-of-life score (QoL), maximum urinary flow rate (Qmax), postvoid residual urine volume (PVR), and prostate and adenoma volume. In all, 62 and 78 patients completed the 1, 2, 3 and 4-year follow-up from the laser and TUVP groups, respectively. RESULTS: At each follow-up, the IPSS, QoL, Qmax and PVR were significantly better and more durable in the TUVP than in the laser group. In the TUVP and laser groups respectively, at the 4-year follow-up the mean value of the IPSS was 3.7 vs 11.9, the QoL 1.3 vs 3.1, the Qmax 21.4 vs 13.6 mL/s and the PVR 25.1 vs 64.6 mL (all P < 0.001). The mean prostate and adenoma volume were significantly lower after TUVP than after laser therapy (P < 0.001) at the 1- and 4-year follow-up, with final values of 27.9 vs 35.9 and 11.7 vs 20 mL (both P < 0.001) for the TUVP and laser groups, respectively. Retrograde ejaculation was significantly more common after TUVP (63%) than after laser therapy (18%; P < 0.001). Impotence was reported in 8% of men after TUVP and in none after laser therapy (P = 0.040). The re-operation rate was 12% after TUVP and 38% after laser treatment (P < 0.001). CONCLUSION: These 4-year follow-up results confirm that TUVP is significantly more effective and durable than the Nd:YAG laser for treating BPH. Residual obstructing adenoma was the main cause of failure in the laser group, which reflects the inadequacy of laser therapy for removing the adenoma. PMID- 12780837 TI - Catheter-associated urinary tract infections: prevalence of uropathogens and pattern of antimicrobial resistance in a UK hospital (1996-2001). AB - OBJECTIVE: To assess the change in the bacterial profile and pattern of antibiotic resistance of catheter-associated urinary tract infections (CAUTIs, the most prevalent form of nosocomial infections) between 1996 and 2001. MATERIALS AND METHODS: Catheter samples of urine (CSUs) submitted in 1996, 1998 and 2001 (2451, 2460 and 3349 specimens, respectively) were analysed. The distribution of different uropathogens in bacterial CAUTIs and their in vitro antimicrobial resistance was evaluated over the study interval. The likelihood ratio test was used to assess whether there was a linear trend according to calendar year. RESULTS: Escherichia coli was the most frequently isolated pathogen in all years, but its frequency declined over time (35.6%, 32.5% and 26.6%, respectively). Enterococcus was the second most frequent overall, with a significant increase in frequency with time (11.8%, 15.3% and 22.0%, respectively). There was also a considerable change in resistance patterns to antibiotics. As a result, in 1996, CAUTIs were least often resistant to ciprofloxacin (8.0%) followed by co-amoxiclav (18.5%) and cephalexin (25.4%). In 2001, CAUTIs were least often resistant to co-amoxiclav (22.5%), followed by ciprofloxacin (27.2%) and nitrofurantoin (28.8%). CONCLUSION: The types of organisms associated with CAUTI have changed over the last 5 years in a UK institution, as have the patterns of antibiotic resistance. Currently, the most appropriate agents for the empirical management of CAUTIs seem to be co amoxiclav, ciprofloxacin and nitrofurantoin. PMID- 12780838 TI - The acute effect of magnetic stimulation of the pelvic floor on involuntary detrusor activity during natural filling and overactive bladder symptoms. AB - OBJECTIVE: To evaluate the effect of magnetic stimulation of the pelvic floor (MSPF) on involuntary detrusor activity observed during natural filling, and on the overactive bladder symptom complex. PATIENTS AND METHODS: Eighteen women with detrusor overactivity on conventional cystometry underwent ambulatory urodynamic monitoring over two filling cycles. Fluid intake was standardized, provocative manoeuvres applied at regular intervals and symptoms documented contemporaneously. During the second filling cycle MSPF was delivered whenever the detrusor pressure increased by > 5 cmH2O. The women were subsequently treated with MSPF for 6 weeks; their lower urinary tract symptoms were assessed before and after treatment. RESULTS: Comparing the second (stimulated) cycle with the first (unstimulated) cycle, cystometric capacity was higher (373 vs 224 mL, P < 0.03). and involuntary detrusor activity of shorter duration (370 vs 427 s, P < 0.82) and lower amplitude (53 vs 63 cmH2O, P < or = 0.05). All women tolerated the procedure comfortably, but nine found it too time-consuming and withdrew. In the nine women who completed treatment there was no consistent change in overactive bladder symptoms. CONCLUSIONS: In this pilot study, MSPF during natural filling was associated with a decrease in the amplitude of involuntary detrusor contractions and a significant increase in cystometric capacity. However, MSPF had a variable effect on sensations of urgency, both acutely and after treatment, and currently there is no evidence to suggest that MSPF has an enduring effect on symptoms of the overactive bladder. PMID- 12780839 TI - A study to detect Helicobacter pylori in fresh and archival specimens from patients with interstitial cystitis, using amplification methods. AB - OBJECTIVES: To detect Helicobacter pylori DNA in fresh and paraffin-embedded bladder biopsy specimens, and thus determine any possible role in interstitial cystitis (IC). MATERIALS AND METHODS: Thirty-three bladder biopsy samples were examined from patients with IC (29 paraffin-embedded and four freshly frozen) diagnosed according to National Institute of Diabetes, Digestive and Kidney Disease criteria. The positive control was a gastric biopsy sample from a patient with a known gastric ulcer caused by H. pylori infection. RESULTS: The anticipated polymerase chain reaction product size of 109 base pairs was obtained with the positive control, whereas none of the other biopsy samples (paraffin embedded or fresh) showed positive amplification specific for H. pylori. CONCLUSIONS: As there was no H. pylori DNA in any of the samples from patients with IC, it is an unlikely candidate in the pathogenesis of IC. PMID- 12780840 TI - Laparoscopic renal biopsy: a 9-year experience. AB - OBJECTIVES: To present our experience and outcome of consecutive laparoscopic renal biopsy over a 9-year period, as renal biopsy remains an important diagnostic procedure for evaluating proteinuria, haematuria and renal failure, but when percutaneous biopsy is contraindicated, a laparoscopic biopsy is an attractive option because it is minimally invasive. PATIENTS AND METHODS: Seventy four patients (29 male, 45 female, mean age 45 years, range 3-79) had a laparoscopic renal biopsy taken for various indications, e.g. morbid obesity, solitary kidney, coagulopathy, failed percutaneous biopsy, high location of the kidney and poor visualization with ultrasonography. The kidney was approached via a laparoscopic retroperitoneal route using a two-port technique, with the patient in the flank position. After identifying the kidney, one to five cortical biopsies were obtained with cup-biopsy forceps. RESULTS: Adequate tissue was obtained in 96% of the patients; the mean (range) operative duration was 123 (9 261) min and the estimated blood loss 67 (5-2000) mL. Forty-three patients were discharged within 24 h. Complications occurred in 10 patients, with significant bleeding in three. One patient died after surgery, secondary to a perforated peptic ulcer while on high-dose steroid therapy. CONCLUSION: Laparoscopic renal biopsy is a safe and effective alternative to open renal biopsy for patients in whom percutaneous biopsy is not feasible. It offers the advantage of obtaining cortical biopsies and achieving haemostasis under direct vision. Adequate renal tissue is obtained in most cases. Recovery and convalescence are short for most patients. PMID- 12780841 TI - Artificial neural networks in predicting optimum renal stone fragmentation by extracorporeal shock wave lithotripsy: a preliminary study. AB - OBJECTIVE: To assess the ability of artificial neural networks (ANNs) to predict optimum renal stone fragmentation in patients being managed by extracorporeal shock wave lithotripsy (ESWL). PATIENTS AND METHODS: The study included 82 patients with renal stones who were being treated by ESWL. Data (input and output values) from 60 patients in whom there was optimum fragmentation of stones by ESWL were used for training the ANN. These data mainly included the 24-h urinary variables, the radiological features of the stone disease and the ESWL settings used. The predictability of the trained ANN was tested on 22 subsequent patients, by supplying the input variables of the 22 patients into the trained ANN and recording the output values (predicted values). After subjecting these patients to ESWL, the actual results (observed values) were recorded. The predicted and the observed values were then compared. RESULTS: In the 22 patients in whom predictability was tested, the trained ANN predicted optimum fragmentation at < or = 13 000 shocks/stone (as per study protocol) in 17 and optimum fragmentation at> 13 000 shocks/stone in the other five. In the 17 patients (test set) where the trained ANN had predicted optimum fragmentation at < or = 13 000 shocks/stone, the optimum fragmentation was at that value, although the predicted and observed values were not identical. The overall correlation between the predicted and the observed values was 75.5% (correlation coefficient 0.7547) in these 17 patients. Of the other five patients, none had optimum fragmentation at < 13 000 shocks/stone, as predicted by the trained ANN, giving complete accuracy for this factor. CONCLUSION: This was a pilot study, i.e. an initial attempt to use an ANN in this regard, and although there were few patients, such that it is not possible to make final recommendations, the overall predictability was approximately 75%. An encouraging outcome of the study was that the trained ANN identified patients unlikely to benefit from ESWL. Using a larger dataset and identifying more significant variables, while eliminating inputs with a negative effect, the efficiency and utility of this ANN can probably be enhanced and in future it might be possible to predict stone fragmentation with reasonable accuracy. PMID- 12780842 TI - Transdermal electromotive administration of verapamil and dexamethasone for Peyronie's disease. AB - OBJECTIVE: To evaluate the effects of the transdermal electromotive administration of verapamil and dexamethasone on plaque size, penile deviation, pain, erectile function and capacity for vaginal penetration in patients with Peyronie's disease. PATIENTS AND METHODS: Forty-nine patients were treated four times weekly for six consecutive weeks. During each session the drug mixture was administered from a receptacle fixed to the skin overlying the plaques, using 2.4 mA pulsed current for 20 min. Plaque size and penile deviation were evaluated by dynamic penile duplex ultrasonography, X-ray and photographs; pain, erectile function and capacity for vaginal penetration were assessed using a questionnaire. Vital signs and side-effects were recorded. Differences before and after treatment were assessed. RESULTS: The plaque disappeared in 8% of patients, with a measurable reduction in volume in 74% and no change in 18% (P < 0.001). Penile deviation resolved in 10% of the men, decreased in 74% and remained unchanged in 16% (P < 0.001). The plaque volume was halved in two-thirds of the men, to a mean (sd) of 515 (301) mm3, and the penile deviation halved in 45% of patients, to 24 (5) degrees; pain was completely eliminated in 88% (P < 0.001). Erectile function was completely restored in 42% of patients with initial erectile dysfunction and improved in 17% (P < 0.001); vaginal penetration improved in 73%. No toxicity was noted, except for a transient skin erythema at the site of the penile and dispersive electrodes. CONCLUSION: The transdermal electromotive administration of verapamil and dexamethasone is clinically safe and appears to be an effective treatment in patients with Peyronie's disease. PMID- 12780843 TI - Fibrin as an inducer of fibrosis in the tunica albuginea of the rat: a new animal model of Peyronie's disease. AB - OBJECTIVES: To investigate the role of fibrin in inducing fibrosis in the tunica albuginea (TA) of the rat penis, to develop a new animal model for Peyronie's disease (PD). MATERIALS AND METHODS: The TA of rats (five per group per period) were injected with either saline, fibrin, transforming growth factor-beta1 (TGF beta1) or TGF-beta1 plus fibrin; the rats were killed at 1, 3, and 6 weeks after injection. Images were analysed quantitatively from tissue sections stained for collagen (Masson trichrome), fibrin (Verhoeff's stain) and elastin (Hart's stain), and immunostained for TGF-beta1, inducible nitric oxide synthase (iNOS), heme oxygenase 1 (HO1), alpha-smooth muscle actin (ASMA), apoptosis (TUNEL) and plasminogen activator inhibitor (PAI). Collagen fibre organization was characterized by electron microscopy. Human PD plaque tissue and normal human TA were assayed for fibrin by immunohistochemistry in nine samples. RESULTS: At 1 week after injection of fibrin into the rat TA, only oedema was present; at 3 weeks, the oedema developed into a characteristic fibrotic PD-like plaque. The injection of TGF-beta1 into the TA also induced oedema in the TA at 1 and 3 weeks but there was very little evidence of a recognisable plaque at either time. Injection with TGF-beta1 plus fibrin resulted in oedema at 1 week but at 3 weeks there was a smaller plaque than with fibrin only. At 6 weeks the induced plaques in the fibrin-only and fibrin + TGF-beta1 groups persisted, and were comparable with those elicited at this time by TGF-beta1 alone. The control animals showed no pathology at any of the sample times. At 3 weeks the PD plaque induced by injection with fibrin alone had not only greater expression of TGF-beta1 than the TA of the animals receiving TGF-beta1 alone, but also greater levels of other markers of fibrosis, e.g. HO1 (reactive oxygen species), ASMA (presence of myofibroblasts), apoptosis, and PAI (inhibitor of fibrinolysis). iNOS, a known antifibrotic agent, was also increased. In human PD plaque tissue, fibrin was detected by immunohistochemistry in all nine specimens. CONCLUSIONS: These results suggest that fibrin, when introduced into the TA of the rat penis, acts as a potential profibrotic protein, possibly via the local release of TGF-beta1, and induces a plaque not only histologically similar to that induced by TGF-beta1 but to that of the human condition. Because fibrin can extravasate from the blood into the human TA after an injury to the TA, and because fibrin persists in the plaque tissue, we hypothesise that fibrin may play a key role in the pathogenesis of human PD. PMID- 12780844 TI - Current practice in the management of vasectomy reversal and unobstructive azoospermia in Merseyside & North Wales: a questionnaire-based survey. AB - OBJECTIVE: To investigate the current incidence of vasectomy reversal procedures, the techniques used and which practitioners use them. PATIENTS AND METHODS: Using a questionnaire, 130 general surgeons and urologists practising in Merseyside and North Wales were surveyed. RESULTS: The response rate was 74%, with 24 urological surgeons and 14 general surgeons undertaking vasectomy reversal. Annually, urological surgeons carried out significantly more procedures than did general surgeons, at 8.5 and 5.3 (P = 0.029), respectively. They were also more likely to use double-layer closure and microsurgical techniques, whilst significantly less likely to use stents. Urologists reported significantly greater patency rates, at 76% and 52% (P = 0.017), respectively, with no significant differences in subsequent pregnancy rates (30% vs 25%). Only one practitioner checked tubal patency in the female partner before vasectomy reversal. CONCLUSIONS: The use of vasectomy reversal is a cost-effective treatment for men wanting paternity after vasectomy. The technique used by the clinician and proper audit of the results require close attention; it would also appear to be obvious that all the partners of men seeking a vasectomy reversal should have their fertility status established before reversal, something that is clearly not done at present. PMID- 12780845 TI - Urogynaecological and obstetric issues in women with the exstrophy-epispadias complex. AB - OBJECTIVE: To review of the sexual and urogynaecological issues faced by a large cohort of women with the exstrophy-epispadias complex (EEC). PATIENTS AND METHODS: The study comprised 83 women and girls with EEC; a confidential survey was mailed to identify their social and sexual concerns. Fifty-six women had classical bladder exstrophy (CBE), 13 had female epispadias (FE) and 14 had cloacal exstrophy (CE). Data on the initial method of reconstruction and urogynaecological problems were obtained from a review of the hospital records. Information on continence, infection and sexual function was obtained from 34 completed surveys. RESULTS: The bladder was closed in 51 patients with CBE and 13 with CE. Urinary calculi developed in 10 patients with CBE, two with FE and three with CE. Vaginal and uterine prolapse occurred an earlier age in patients with EEC. Eight women had 13 pregnancies, eight of which resulted in normal healthy children. Overall continence was achieved in 85% of the women surveyed. Urinary tract infections remained a frequent problem for women with EEC; only 27% of respondents indicated that they were infection-free. Women aged > 18 years (24) who responded indicated that they had appropriate sexual desire; 16 were sexually active and the mean age for commencing sexual activity was 19.9 years. Six patients had dyspareunia and 10 indicated that they had orgasms. However, five additional patients indicated that they had restricted intercourse, as they were dissatisfied with the cosmesis of their external genitalia. CONCLUSIONS: Sexual and gynaecological issues become increasingly important in patients with EEC as they become adults. Understanding these issues faced by patients with EEC as they mature will permit better counselling of future patients. PMID- 12780846 TI - The results of 15 years of consistent strategy in treating antenatally suspected pelvi-ureteric junction obstruction. AB - OBJECTIVE: To determine how to select patients for surgery among those with antenatally detected pelvi-ureteric junction (PUJ) obstruction. PATIENTS AND METHODS: The study comprised 100 consecutive children with antenatally detected suspected unilateral PUJ obstruction and a normal contralateral kidney. The correct diagnosis was made using postnatal ultrasonography, intravenous urography and renal scintigraphy, the last also being used for the follow-up. RESULTS: Four patients had poor function in the hydronephrotic kidney, treated in three by nephrectomy; 61 had normal function in the hydronephrotic kidney, with 49 followed for 1-10 years with no change in kidney function and no symptoms. Twelve patients in this group had later surgery (at 0.7-8 years old) because of pyelonephritis (four), pain and/or renal functional impairment (eight, three of whom had normal function afterward). Thirty-five patients had moderately impaired function of the hydronephrotic kidney; 29 had primary surgery at a median age of 4 months. The median hydronephrotic renal function increased from 32% before to 42% after surgery, with 15 kidneys having normal function. In one other patient the kidney was lost before surgery. Five other patients were initially treated conservatively and the hydronephrotic renal function increased from 32% to 35% at the 1-year follow-up, significantly less than in the surgical group. The overall operative complication rate was 4%. CONCLUSIONS: With our management programme more than half the patients can avoid surgery in childhood. Although the follow up was intense there was moderate and irreversible functional kidney deterioration in 5%. In contrast, after successful reconstructive surgery, only a few follow-up procedures causing possible discomfort to the child are needed in most. The risk of surgical complications cannot be neglected. The present results are useful for advising parents deciding whether their antenatally detected hydronephrotic child should undergo surgery or not. PMID- 12780847 TI - The long-term outcome of bilateral Cohen ureteric reimplantation under a common submucosal tunnel. AB - OBJECTIVE: To analyse the results of bilateral Cohen reimplantation under a common submucosal tunnel, over an 18-year period. PATIENTS AND METHODS: We retrospectively examined 102 children (35 boys and 67 girls, median age 5.5 years, range 0.5-13.5) who underwent bilateral antireflux ureteric reimplantation from 1983 to 2000 with a modified Cohen technique, re-implanting both ureters under a common submucosal tunnel in the mid-trigonal area, to treat primary vesico-ureteric reflux (VUR, 99 patients) or obstructive megaureter (three). The mean (range) follow-up was 10.6 (2-18) years. RESULTS: The operation was successful in 198 of 204 (97%) ureters. One patient had vesico-ureteric stenosis in one ureter and was re-operated successfully. In two ureters in two different patients there was transient stasis after surgery caused by oedema within the tunnel, which gradually resolved. Two ureters in two other patients had reflux after surgery, which resolved spontaneously after 12 and 24 months, respectively. A 6-month old baby had anuria after surgery because of acute compression of both ureters within a narrow tunnel; this patient was re-operated, the tunnel widened and the obstruction resolved. None of 82 patients who had reached school age by the time of their last follow-up showed signs of voiding dysfunction. CONCLUSIONS: The modified bilateral Cohen reimplantation with both ureters under a common submucosal tunnel offers very good long-term results in curing VUR or obstructive megaureter. Crossing one ureter upon the other within the tunnel does not predispose to long-term obstruction. From these results we recommend it as a reliable technique for surgically treating bilateral VUR or obstructive megaureter. PMID- 12780848 TI - Detection of vesico-ureteric reflux in children by colour-flow Doppler ultrasonography. AB - OBJECTIVES: To assess the findings of colour-flow Doppler ultrasonography (DUS) in the diagnosis of vesico-ureteric reflux (VUR), as an alternative to the primary diagnostic tool, voiding cysto-urethrography (VCUG). PATIENTS AND METHODS: Thirty-five children (aged 2-15 years) were examined for VUR by DUS and standard VCUG. All patients underwent DUS and VCUG within 48 h but the findings of the latter were not reported to the ultrasonographer. The DUS was undertaken using a colour Doppler real-time system; a positive ultrasonogram was defined by visualizing Doppler signals from the bladder to the ureter during the course of bladder filling. RESULTS: Of the 70 ureters assessed, 28 were refluxing on DUS and 29 on VCUG; there were two false-positive and three false-negative results. Comparing the two methods showed DUS to be 90% sensitive with a specificity of 93% for detecting VUR. Four patients underwent reimplantation during their treatment. DUS findings correlated well with standard VCUG in these patient as a method of follow-up. CONCLUSION: DUS can be used as an alternative to standard VCUG for screening and following VUR. In addition, DUS avoids the danger of exposure to ionizing radiation and the unpleasant catheterization that many of these children fear. PMID- 12780849 TI - Urodynamics in a rat neurogenic bladder model with a unilateral electrolytic lesion of the basal forebrain. AB - OBJECTIVE: To investigate the changes in bladder function in rats with an electrolytic lesion of the right basal forebrain (RBF) and to determine the effects of AH-9700, a novel sigma receptor ligand, on cystometry in RBF-lesioned rats. MATERIALS AND METHODS: A lesion was made electrolytically in the RBF of male Wistar rats. At 7 or 8 days after the lesion or sham surgery, continuous cystometry was performed in awake rats. In addition, contractile responses to electrical field stimulation or carbachol were measured in isolated bladder strips, as were the forebrain contents of acetylcholine, monoamine neurotransmitters and their metabolites. RESULTS: RBF-lesioned rats showed a remarkable increase in voiding frequency, with a decrease in voiding threshold pressure but no change in voiding pressure, compared with sham-operated rats. However, contractile responses in bladder strips isolated from RBF-lesioned rats were no different from those in strips isolated from sham-operated rats. In RBF lesioned rats, the contents of acetylcholine, dopamine, 4-dihidroxyphenylacetic acid and homovanillic acid were significantly decreased in the right forebrain. AH-9700 dose-dependently decreased the voiding frequency and increased the threshold pressure in RBF-lesioned rats. Anti-muscarinic agents (oxybutynin and propiverine) also decreased the voiding frequency, but their effects were less potent than that of AH-9700. CONCLUSIONS: The RBF-lesioned rat may be a useful model for the neurogenic bladder of supraspinal origin. Moreover, AH-9700 effectively improves bladder dysfunction in this model. PMID- 12780850 TI - Spontaneous regeneration of the seriously injured sympathetic pathway projecting to the prostate over a long period in the dog. AB - OBJECTIVES: To explore the spontaneous regeneration, over a long period, of the seriously injured sympathetic pathway controlling the prostate. MATERIALS AND METHODS: The hypogastric nerve (HGN), which is part of the sympathetic pathway from the spinal cord to the prostate, was partly removed over half of its length on both sides in six dogs. Four years after surgery the responses of the prostate to electrical stimulation of the lumbar splanchnic nerve (LSN) or the HGN proximal to the site removed, were assessed. RESULTS: In six dogs, 10 of the 17 LSNs (second to fourth) and four of the 10 HGNs stimulated elicited prostatic contraction. The pathways via the ipsilateral HGN and/or the contralateral HGN from the LSNs to the prostate were identified as having regenerated in four of six dogs. CONCLUSION: These results indicate that the sympathetic pathways via the HGN to the canine prostate can regenerate spontaneously over a long period after serious injury, and that their cross-innervation system can also be repaired. PMID- 12780851 TI - Comparison of the relaxant effects of alfuzosin, phentolamine and sildenafil on rabbit isolated corpus cavernosum. AB - OBJECTIVE: To compare the direct relaxant effects of alfuzosin, phentolamine and sildenafil in rabbit isolated corpus cavernosum (CC) pre-contracted with phenylephrine or KCl. MATERIALS AND METHODS: Penile erectile tissue was obtained from male New Zealand White rabbits (22-26 weeks old). The CC was cut into longitudinal strips and mounted under 2 g resting tension in 5-mL jacketed organ baths containing a modified Krebs solution bubbled with 95% O2, 5% CO2 and maintained at 37 degrees C. Tissue strips were pre-contracted by 60 mmol/L KCl or 10 micro mol/L phenylephrine. After obtaining a stable plateau of contractions, test compounds were added to the organ bath. The relaxant potencies were expressed as the percentage of inhibition of the plateau of contraction induced by 10 micro mol/L phenylephrine. RESULTS: Alfuzosin showed a concentration dependent relaxing effect on rabbit CC pre-contracted by 10 micro mol/L phenylephrine, with a mean (sd) pIC50 of 7.64 (0.06). The relaxant effect was unaffected by pre-incubation with 100 micro mol/L Nomega-nitro-l-arginine methyl ester (L-NAME). Phentolamine had a potency similar to alfuzosin, with a pIC50 of 7.44 (0.08). Both alfuzosin and phentolamine were completely ineffective on the plateau of contraction induced by 60 mmol/L KCl. In contrast to alfuzosin, sildenafil was equipotent in relaxing the rabbit CC against each contractile agent, with pIC50 values of 7.25 (0.09) and 7.23 (0.22) with 10 micro mol/L phenylephrine and 60 mmol/L KCl, respectively. The relaxant response to sildenafil was partly blocked by pretreatment with 100 micro mol/L L-NAME, with pIC50 values of 7.94 (0.09) and 6.63 (0.32) without and with L-NAME, respectively. Sildenafil, incubated for 45 min at 10 micro mol/L, had no relaxant effect on the resting tension of the preparation or on the concentration-response curve to phenylephrine. CONCLUSIONS: The direct relaxant effect of alfuzosin is mediated through alpha1-adrenoceptor blockade. The relaxations induced by phentolamine and alfuzosin are independent of nitric oxide, whereas those induced by sildenafil are, at least partly, sensitive to L-NAME and a selective soluble guanylate cyclase inhibitor, indicating the involvement of nitric oxide and soluble guanylate cyclase. Alfuzosin and phentolamine effectively counteract alpha1-adrenoceptor-mediated contractions of rabbit CC. If valid for human CC, such an effect may contribute to an improved erectile function in patients treated for benign prostatic hyperplasia. PMID- 12780852 TI - Structural and ultrastructural studies of the urinary tract of mice inoculated with Lactobacillus fermentum. AB - OBJECTIVE: To assess, using structural and ultrastructural studies of the urinary tract, the effects of the intraurethral inoculation of lactobacilli (probiotic treatment) as lactobacilli are the predominant micro-organisms of the urogenital tract of humans, monkeys and mice. MATERIALS AND METHODS: Previous work showed the protective effect of Lactobacillus fermentum CRL 1058 intraurethrally inoculated against the challenge of uropathogenic Escherichia coli. There was also an effect of oestrogens and antibiotics in the kinetics of colonization of both micro-organisms in mice. In the present study L. fermentum was inoculated with agarose beads (107 colony-forming units) and the number of micro-organisms determined by plating in selective media, giving a high degree of colonization in all the organs studied. The urinary tract organs were processed by histological and electron microscopy techniques standardized in our laboratory. RESULTS: The intraurethral inoculation of lactobacilli produced no adverse effects or significant changes in any of the organs assessed (kidney, ureter, bladder or urethra), when evaluated by histological and ultrastructural techniques. CONCLUSION: The use of lactobacilli as a probiotic treatment is probably safe. PMID- 12780853 TI - ADAM and the andropause. PMID- 12780854 TI - A novel method for laparoscopic placement of Tenckhoff peritoneal dialysis catheters. PMID- 12780855 TI - A digital rectal examination after radical prostatectomy is unnecessary if prostate specific antigen is undetectable. PMID- 12780856 TI - Primary T-cell rich B-cell lymphoma of the penis: a first case. PMID- 12780857 TI - Right common iliac artery pseudoaneurysm-ureteric fistula. PMID- 12780858 TI - Congenital seminal vesicle cyst: prenatal diagnosis and postnatal laparoscopic excision with an attempt to preserve fertility. PMID- 12780859 TI - Pregnancy and vaginal delivery after augmentation cystoplasty. PMID- 12780861 TI - Prostatectomy - a radical approach. PMID- 12780862 TI - Hand-assisted laparoscopic nephrectomy: complications related to the hand-port site. PMID- 12780863 TI - The mesh wallstent in the treatment of detrusor external sphincter dyssynergia in men with spinal cord injury: a 12-year follow-up. PMID- 12780864 TI - Spermatogenesis and spectral echo-colour Doppler traces from the main testicular artery. PMID- 12780870 TI - Statistical methods for longitudinal research on bipolar disorders. AB - OBJECTIVES: Outcomes research in bipolar disorders, because of complex clinical variation over-time, offers demanding research design and statistical challenges. Longitudinal studies involving relatively large samples, with outcome measures obtained repeatedly over-time, are required. In this report, statistical methods appropriate for such research are reviewed. METHODS: Analytic methods appropriate for repeated measures data include: (i) endpoint analysis; (ii) endpoint analysis with last observation carried forward; (iii) summary statistic methods yielding one summary measure per subject; (iv) random effects and generalized estimating equation (GEE) regression modeling methods; and (v) time-to-event survival analyses. RESULTS: Use and limitations of these several methods are illustrated within a randomly selected (33%) subset of data obtained in two recently completed randomized, double blind studies on acute mania. Outcome measures obtained repeatedly over 3 or 4 weeks of blinded treatment in active drug and placebo sub-groups included change-from-baseline Young Mania Rating Scale (YMRS) scores (continuous measure) and achievement of a clinical response criterion (50% YMRS reduction). Four of the methods reviewed are especially suitable for use with these repeated measures data: (i) the summary statistic method; (ii) random/mixed effects modeling; (iii) GEE regression modeling; and (iv) survival analysis. CONCLUSIONS: Outcome studies in bipolar illness ideally should be longitudinal in orientation, obtain outcomes data frequently over extended times, and employ large study samples. Missing data problems can be expected, and data analytic methods must accommodate missingness. PMID- 12780871 TI - Treatment-latency and previous episodes: relationships to pretreatment morbidity and response to maintenance treatment in bipolar I and II disorders. AB - OBJECTIVE: To clarify relationships of treatment delay and pretreatment episode count with pretreatment morbidity and responses to maintenance treatments in bipolar disorders. METHODS: In 450 DSM-IV bipolar I (n = 293) or II (n = 157) patients (280 women, 170 men), we evaluated correlations of latency from illness onset to starting maintenance treatment and pretreatment episode counts with pretreatment morbidity and treatment response. We considered morbidity measures before and during treatment, and their differences. RESULTS: Latency averaged 7.8 years, with 9.0 episodes per patient, before various maintenance treatments started. Morbidity (percentage of time-ill, episodes per year, first wellness interval, or proportion of subjects hospitalized or having no recurrences) during maintenance treatment averaging 4.2 years was unrelated to treatment latency or pretreatment episode count. However, pretreatment morbidity was greater with shorter latency, resulting in larger relative reduction of morbidity after earlier treatment. CONCLUSIONS: Greater treatment latency and pretreatment episode count were not followed by greater morbidity during treatment, although longer delay yielded smaller during-versus-before treatment reduction in morbidity. Predictions that longer treatment delay or more pretreatment episodes lead to poorer responses to various maintenance treatments in bipolar I or II disorder were not supported. PMID- 12780872 TI - The effect of lithium in bipolar disorder: a review of recent research evidence. AB - Recently published clinical research on lithium is briefly reviewed. The antimanic effect of lithium is supported by recent evidence. It is confirmed that a drastic reduction of affective morbidity is very frequent in bipolar patients receiving lithium prophylaxis regularly for several years, but that the impact of prophylaxis on the course of bipolar disorder is significantly limited by the high drop-out rate. Lithium does seem to be efficacious also in bipolar disorder with mood-incongruent psychotic features or with rapid cycling. The effect of lithium prophylaxis does not seem to decrease over time, at least in the large majority of patients. The recurrence risk is increased in the months following lithium discontinuation. Lithium seems to exert an antisuicidal effect in bipolar patients. PMID- 12780873 TI - Valproate. AB - OBJECTIVES: This article summarizes the role of valproate as a treatment for bipolar disorder and related conditions. METHODS: Published studies and reviews were systematically reviewed. Results from randomized, parallel group, double blind, placebo-controlled studies that included an active comparator are emphasized. RESULTS: Valproate is an effective treatment for manic patients. Valproate was superior to placebo in one 1-year randomized, parallel group study in rate of recurrence requiring discontinuation, rate of depression requiring discontinuation, total early termination and time to 25% of patients relapsing with mania, and in controlling mild depressive symptoms. On some measures, including time to development of a manic episode, valproate did not differ from placebo. Assessments of maintenance efficacy of valproate and other putative prophylactic treatments for bipolar disorder are problematic, because of the need to analyze multiple indices of efficacy, and practical and ethical issues that limit generalizability of results of placebo-controlled studies. Valproate has some advantages over lithium in treatment of mania for persons with more severe illnesses. Valproate benefits a broader spectrum of bipolar conditions than lithium. Valproate appears at best modestly effective for bipolar depression. Used in combination with several other treatments, additive benefits result, that are greater than with any of the treatments as monotherapy. Side effects are generally mild and manageable, particularly with divalproex. Weight gain and pharmacokinetic interaction with lamotrigine are perhaps the most consistent problems in use. Valproate contributes to neural tube defects if taken during the first trimester of pregnancy, and this risk must be conveyed to women. CONCLUSIONS: Valproate is an effective and useful treatment for bipolar disorder. Studies clarifying its spectrum of efficacy, its safety and efficacy in combination regimens, and its mechanisms of action are warranted. PMID- 12780875 TI - Combination treatment in bipolar disorder: a review of controlled trials. AB - OBJECTIVES: Monotherapy is often inadequate and combination drug regimens have become the norm for the treatment of bipolar disorder. Virtually all classes of psychotropic drugs have been used in bipolar disorder in combination for a variety of indications. This article reviews the available published data from controlled, blinded studies regarding combination treatments in the different treatment phases of bipolar disorder. METHODS: Articles for this review were obtained from a search of the Medline database (1966-2002), using the following keywords and phrases: add-on, antipsychotic, anticonvulsant, antidepressant, combination treatment, lithium, neuroleptic, and polypharmacy. The search was augmented by data presented at scientific meetings. Data included in this article were only from controlled studies that evaluated combinations of two or more agents. RESULTS: For acute mania, the most useful combination treatments as determined by controlled studies, appear to be an antipsychotic drug with a mood stabilizer. The combination of lithium and valproate, even though widely used for acute mania, is lacking in controlled data. For acute bipolar depression, the controlled combination studies reviewed fail to show clear advantages in efficacy of an antidepressant with a mood-stabilizer versus two stabilizers or a mood stabilizer alone. Large, controlled, randomized, long-term studies with modern antidepressants are not available. Controlled combination studies of mood stabilizers suggest gains in efficacy over monotherapy in the long-term treatment of bipolar disorder. CONCLUSIONS: Controlled combination studies in bipolar disorder are uncommon. Increased attention should be given to study combination treatments in all phases of bipolar illness to determine the most efficacious and safest combinations. PMID- 12780874 TI - Alternatives to lithium and divalproex in the maintenance treatment of bipolar disorder. AB - OBJECTIVES: The role of lithium carbonate in the maintenance treatment of bipolar disorder is well established. Unfortunately, many patients fail to respond adequately to this agent or are unable to tolerate its adverse effects. Divalproex has become a commonly used alternative to lithium, but it also is ineffective or poorly tolerated in many patients. This article attempts to review the available data on maintenance therapy in bipolar disorder with a variety of anticonvulsants and antipsychotics (both conventional and novel), with reference to relevant studies in acute mania and bipolar depression as well. METHODS: Evidence on maintenance therapy and relevant acute-phase data were collected using MEDLINE database searches. RESULTS: Data on maintenance therapy with agents other than lithium and divalproex are sparse, and often derived from open, uncontrolled studies. Implications and flaws of available data are discussed. CONCLUSIONS: Other than lithium, there are few robust double-blind data to support the use of a variety of agents in the maintenance phase. However, uncontrolled data suggest that a number of agents merit further study. PMID- 12780876 TI - How rare is bipolar disorder not otherwise specified? PMID- 12780877 TI - Electroencephalogram changes and its improvement with sodium valproate in a patient with electrocution-induced chronic mania. PMID- 12780878 TI - Pilonidal sinus: is surgery alone enough? PMID- 12780879 TI - Pathological staging of epidermoid anal carcinoma for the new era. AB - Chemoradiotherapy is the standard treatment for most patients with epidermoid anal cancer. Pre-treatment staging is based on size for T1-T3 lesions and clinical and radiological assessment of adjacent organ invasion for T4 lesions. For patients with residual or recurrent carcinoma, anorectal excision offers the best chance of oncological salvage. Pathological staging systems for anorectal excision specimens were validated at the time when surgical treatment was first line therapy. A validated staging system is necessary for salvage surgical excision specimens following an attempt to cure by radiotherapy and chemotherapy for the purpose of prognosis and further treatment planning. PMID- 12780880 TI - Quality of life after rectal excision. PMID- 12780881 TI - The rhomboid flap for pilonidal disease. AB - INTRODUCTION: There have been many surgical techniques described for the treatment of pilonidal sinuses. Recurrent disease causes significant morbidity particularly with time from work. AIM: To assess the rhomboid flap's role in promoting one-stage primary healing in pilonidal disease and to evaluate the morbidity and recurrence. METHODS: Fifty-three patients were prospectively recruited of which 27 had previous multiple abscess formation requiring surgical drainage from their pilonidal disease, although none had acute disease at the time of surgery. By using the transposition flap, we were able to obliterate the natal cleft and therefore the rolling action of the buttocks between the cleft in these patients and thereby remove one of the factors involved in pilonidal disease. Hospital stay, healing time, wound infection, wound breakdown and recurrence were noted. RESULTS: There were 47 males and 6 females with a median age of 28 years (range 16-64 years). Median follow-up was 24 months (range 3-36 months). Post-operative morbidity involved superficial wound infection in 7 (13%) which settled with out-patient dressings. There were four recurrences (7%), two occurred between the flap and the anal canal, and the other two in the flap margin needing intervention. All the patients healed their wounds and the median healing time was 14 days. CONCLUSION: As this condition affects a predominantly young population causing significant time off from work, we feel that the Rhomboid Flap is useful for difficult cases in that it allows early return to full activity and does not necessitate prolonged postoperative care. PMID- 12780882 TI - Efficacy and tolerance of a new gentamicin collagen fleece (Septocoll) after surgical treatment of a pilonidal sinus. AB - OBJECTIVE: Pilonidal sinus is common. It causes substantial loss of working hours. The major disadvantage of open wound treatment is the long time required for healing. Primary wound closure is on the other hand often followed by infection. A controlled, multicentre trial was carried out to evaluate the efficacy of a new gentamicin collagen fleece (Septocoll) combined with primary closure. PATIENTS AND METHODS: One hundred and three patients (88 men, 15 women, median age 30 years) were included. Fifty-one were randomised to gentamicin fleece plus primary closure (Genta Group), and 52 patients to open treatment alone (Open Group). RESULTS: The median interval to wound healing was 17 days in the Genta group and 68 days in the Open group (P = 0.0001, log-rank test). Two patients in Group 1 developed infection within the first two weeks, requiring reopening of the wound, with primary wound healing occurring in 73%. Failure of primary healing (27%) was usually due to seroma or spontaneous dehiscence which subsequently healed. CONCLUSION: The combination of gentamicin collagen fleece (Septocoll) with primary closure resulted in a shorter period to healing than the open technique without unwanted effects. PMID- 12780883 TI - The correlation between quality of life and functional outcome in ulcerative colitis patients after proctocolectomy ileal pouch anal anastomosis. AB - OBJECTIVE: The aim of this study was to evaluate functional outcome and quality of life (QOL) in patients undergoing proctocolectomy ileal pouch anal anastomosis (IPAA), to assess the correlation between functional outcome and QOL, and to identify factors influencing functional outcome and QOL in these patients. BACKGROUND: IPAA is now considered the procedure of choice for ulcerative colitis. Functional outcome and QOL are important factors in evaluating operative outcome. METHODS: All patients with UC who had undergone IPAA at our institute during the period 1990-2001 were included. QOL and functional outcome were evaluated by mailed questionnaires. QOL was scored using the Short Form 36 (SF 36). Global Assessment of Function Scale was used to evaluate functional outcome. RESULTS: Data were obtained in 77 of 99 patients (78%), with the median age of 38 years. Median follow up time was 4.25 years. The QOL in patients after pelvic pouch procedure was excellent, with scores equal to published norms for the Israeli general population in most scales. Functional outcome and QOL scores correlated strongly (r > 0.5; P < 0.0001) in all dimensions. Older age was associated with lower scores in both functional outcome and QOL scales (P < 0.0001). CONCLUSIONS: This study demonstrates a strong association between functional outcome and QOL in patients after IPAA. These patients, however, have a QOL that is comparable with the general population. Age at time of surgery strongly influences both functional outcome and QOL. This finding has to be taken into consideration in pre-operative counseling. PMID- 12780884 TI - Colonoscopic surveillance after curative colorectal resection: results of an empirical surveillance programme. AB - INTRODUCTION: Colonoscopic surveillance after colorectal cancer resection is widely practised despite little evidence that it improves survival. The optimum protocol for colonoscopic follow-up after colorectal cancer resection has not yet been elucidated. We audited the outcome of an empirical colonoscopic follow-up programme in a cohort of patients who underwent colorectal resection with a minimum of five years follow-up to establish patterns of metachronous neoplasia and suitable surveillance intervals. METHODS: The colonoscopic records, biopsy results and follow-up details of patients diagnosed with colorectal cancer between June 1990 and June 1996 were reviewed. The number and type of metachronous neoplastic lesions diagnosed was recorded. Rates of development of new neoplasms were estimated by calculating the time from operation to their first discovery. Factors predictive of further development of polyps or cancer were sought. Results were compared to published reports of intensive follow-up programmes. RESULTS: Seven hundred and ninety-eight patients underwent colorectal resection with curative intent during the study period. 226 patients had one or more follow-up colonoscopies (mean time post resection 48.8 months). In total 352 colonoscopies, encompassing 1437 patient years of surveillance, were performed. Nine metachronous cancers in eight patients, five of which were asymptomatic were diagnosed by colonoscopy at a mean of 63 months. Three asymptomatic recurrences were diagnosed but all were inoperable. 70 (31%) patients had adenomatous polyps diagnosed after a mean time from operation of 34 months for simple adenomatous polyps and 21 months for those with advanced features. Patients with multiple polyps or advanced polyps at the initial colonoscopy were more likely to form subsequent polyps. Only 5.8% of patients with a single adenoma or a normal colon formed an advanced adenoma over the next 36 months of surveillance. CONCLUSION: The results of an empirical colonoscopic follow-up programme compared favourably to the results of the intensive programmes reported in the literature. Most patients are at very low risk of developing significant colonic pathology over the first five years after resection. Colonoscopic surveillance intervals need not be less than five years unless the patient has multiple adenomas or advanced adenomas at the first colonoscopy. Three yearly surveillance intervals are most probably adequate in these individuals. PMID- 12780885 TI - Caecal-diverticulitis: a rare differential diagnosis for right-sided lower abdominal pain. AB - OBJECTIVE: There are several differential diagnoses for acute abdominal discomfort with pain referred to the right lower quadrant region. The objective of this report is to outline our experience with diverticulitis of the caecum, which is a rare condition that is particularly difficult to diagnose correctly pre-operatively. METHODS: A retrospective analysis of acute diverticulitis of the caecum was performed over the period from January 1992 to December 2000 within the Surgical Department of the RWTH Aachen, Germany. The progress of each patient was assessed by interviewing the patient and by telephone contact with the patient's general practitioner. RESULTS: Seven patients with isolated infective diverticulitis of the caecum were encountered in the course of this study. In five of these cases, the pre-operative diagnosis was either unclear or incorrect. Four patients underwent surgery for suspected appendicitis. In one patient, the diverticulitis was diagnosed during laparoscopic surgery undertaken for presumed adnexal disease. Intestinal continuity was restored in all cases. The postoperative course was uneventful. CONCLUSIONS: Right-sided caecal diverticulitis is rare in comparison to diverticulitis of the sigmoid or descending colon. Nevertheless, it should be actively considered in the differential diagnosis of patients with acute tenderness in the right lower abdominal region. PMID- 12780887 TI - The influence of bladder filling on anorectal function. AB - OBJECTIVE: The aim of this study was to develop a technique to simultaneously evaluate bladder and anorectal function. In particular, this study was designed to determine if anal sphincter resting pressure, anal sphincter squeeze pressure and rectal sensation change with bladder filling. PATIENT AND METHODS: A pilot study of ten female patients who presented to the pelvic physiology unit for assessment of urinary symptoms was performed. All patients completed a symptom questionnaire and quality of life assessment form. Following informed consent a baseline urodynamic test was performed with the bladder empty and subsequently followed by an anorectal manometric test. Changes in anal sphincter resting pressure, squeeze pressure and rectal pressure were recorded over a ten-minute period. With the patient lying in the left lateral position, the bladder was then filled with isotonic saline at room temperature at a constant rate of 30 ml/min. A continuous assessment of changes in anal sphincter resting pressure during bladder filling was made. Anal sphincter squeeze pressure and rectal sensation were measured at fixed intervals during bladder filling (50, 100, 150 ml etc.) and at fixed intervals relative to bladder capacity (25, 50, 75 and 100% capacity) by stopping bladder filling at the appropriate level. RESULTS: There was no significant change in anal sphincter resting pressure (Mean difference(s.d.) between bladder full and empty = 2.7(5.6) P = 0.92*), squeeze pressure (Mean(s.d.) difference = 9.5(26.3) P = 0.86*) and rectal sensation (Mean difference(s.d.) first sensation 10(15.2) P = 0.958; Mean difference(s.d.) urgency = 10(17.8) P = 0.07*) on bladder filling. CONCLUSION: Under normal physiological circumstances, bladder filling does not influence anorectal function. * STATISTICAL ANALYSIS: Wilcoxon signed rank sum test. P < 0.05 considered significant. Units = mmHg PMID- 12780886 TI - Rectal endosonography can distinguish benign rectal lesions from invasive early rectal cancers. AB - OBJECTIVE: To determine whether an experienced ultrasound examiner, using good ultrasound equipment with high multifrequency probes, can discriminate between a high grade or low grade dysplastic adenoma (pT0) and very early invasive rectal cancers (pT1). SUBJECTS AND METHODS: Sixty consecutive patients with clinically possibly pT0 or pT1 rectal tumours referred for transanal local excision underwent endorectal ultrasound examination. Lesions where the endorectal ultrasound image showed the mucosal layer to be expanded but the submucosal layer to be intact (uT0) were considered to represent a low grade or high grade dysplasia adenoma (pT0). An irregularity or disruption of the submucosal layer (uT1) was considered to characterize early invasive rectal cancers (pT1). The ultrasound staging was compared with the histological staging made on the basis of the diagnoses in the excised specimens. RESULTS: The histopathological diagnoses were: invasive rectal cancer (n = 18, 10 pT1, 4 pT2, 4 pT3 cancers); high grade dysplastic adenoma (n = 21); low grade dysplastic adenoma (n = 18); non adenomatous benign lesions (n = 3). Endorectal ultrasound incorrectly classified two of the invasive cancers (both pT1 tumours) as noninvasive lesions. Five of 42 pT0 tumours were overstaged as uT1 tumours. Overstaging was more common in patients who had undergone a previous excision and in tumours with peritumoral inflammation and desmoplastic reaction. The sensitivity of endorectal ultrasound with regard to invasive cancer was 89% (16/18), specificity 88% (37/42), positive predictive value 76% (16/21), negative predictive value 95% (37/39), and accuracy 88% (53/60). Among pT0 and pT1 tumours, the corresponding figures were 80% (8/10), 88% (37/42), 62% (8/13), 95% (37/39), and 87% (45/52). CONCLUSION: Endorectal ultrasound can distinguish between noninvasive lesions and invasive rectal cancers clinically of stage pT0 or pT1. PMID- 12780888 TI - A prospective randomized trial of diltiazem and glyceryltrinitrate ointment in the treatment of chronic anal fissure. AB - OBJECTIVE: The aim of this study was to compare prospectively diltiazem with GTN ointment in the treatment of anal fissure. PATIENTS AND METHODS: Of 43 outpatients with chronic anal fissure, 22 patients were randomized to topical diltiazem (2%) ointment and 21 patients to glyceryltrinitrate (GTN) (0.5%) ointment twice daily for 8 weeks. During the course of treatment each patient was seen three times. Side-effects and healing were recorded. RESULTS: Healing occurred in 19 of 22 patients treated with diltiazem and 18 of 21 patients were cured with GTN (P = 0.95). Those who were treated with nitroglycerin ointment developed headache and dizziness developed after GTN in 33.3% of cases while no patient had any side-effects after diltiazem. CONCLUSIONS: Diltiazem and glyceryltrinitrate (GTN) were equally effective in healing anal fissure but the former resulted in fewer side-effects. PMID- 12780889 TI - Anorectal function following low anterior resection. AB - INTRODUCTION: Rectal function following rectal resection for carcinoma was studied in 43 patients. METHODS: Sixteen women and 27 men with a median age of 66 years (range 41-79 years) were included. Twenty-three patients had a diverting ileostomy at the time of resection. Eight patients had a 6-cm colonic J-pouch. Ten patients had anastomotic leakage including two patients without diverting ileostomy. One patient had pre-operative radiation with 25 Gy. The patients were studied at a median 12 months (range 3-30) after rectal resection. Distance from anal verge to the anastomosis was a median 7 cm (range 3-12 cm). Clinical data, anal manometry and rectal compliance were analysed. RESULTS: Stool frequency was 3 per day (range 1-10). Twenty-two (51%) patients were continent, 11 (26%) were incontinent for flatus, and 10 (23%) were incontinent for faeces (three for liquid and seven for solid stool). Fourteen (33%) patients had constipation, two of whom also had incontinence for solid or liquid stool. The level of the anastomosis for patients with postoperative constipation was 5 cm (range 3-12 cm), while it was 7 cm (range 3-10 cm) for nonconstipated patients (NS). Anal manometry was normal. Rectal compliance was lower in patients with incontinence for liquid or solid faeces than in patients with flatus incontinence only (P < 0.01), and rectal volume tolerability was lower in incontinent patients compared with continent patients (P < 0.05). The rectoanal reflex was present in 31 (72%) patients. There was a negative correlation between maximal rectal volume and stool frequency and between level of the anastomosis and degree of incontinence. Age did not affect functional outcome. CONCLUSION: Many patients had a poor functional result following low anterior resection. One in four suffered from incontinence to liquid or solid faeces and one third of the patients experienced constipation. A low level of anastomosis tended to increase stool frequency and carried a higher risk of incontinence. Patients with faecal incontinence tended to have lower rectal compliance and volume tolerability than patients who were continent, while there was no difference in anal pressures. PMID- 12780890 TI - Safety of primary anastomosis in emergency colo-rectal surgery. AB - BACKGROUND: The surgical management of left-sided large bowel emergency patients remains controversial. There has been an increasing trend towards primary reconstructive surgery. The main dilemma remains appropriate patient selection for primary anastomosis. METHODS: The records of 323 patients who presented as acute emergencies and underwent surgery between January 1990 and December 2000 for left-sided colorectal cancer and diverticular disease were reviewed, to compare the outcome of resection and primary anastomosis with Hartmann's procedure. Patients were stratified into 3 groups according to whether the presentation was with localized or generalized peritonitis, or with obstruction. RESULTS: Resection and anastomosis was carried out in 176 (55.7%) patients with a 30-day mortality of 5.7%. Anastomotic dehiscence occurred in 9 (5.1%) patients, with no difference between the three groups. Wound sepsis occurred in 8 (4.5%) patients, and the median hospital stay was 13 days. Hartmann's resection was associated with a higher incidence of systemic and surgical morbidity (39.5% and 24.3%, respectively). The mortality rates in those selected for primary anastomosis (5.7%) compared favourably with those undergoing Hartmann's resections (20.4%) (P < 0.001). CONCLUSION: Emergency primary anastomosis in left sided disease can be performed with a low morbidity and mortality in selected patients, even in the presence of a free perforation with diffuse peritonitis. Patients selected for staged resection, were those with major comorbid disease. PMID- 12780891 TI - A novel indicator for surveillance colonoscopy following colorectal cancer resection. PMID- 12780892 TI - A novel indicator for surveillance colonoscopy following colorectal cancer resection. PMID- 12780893 TI - Further cautionary tales from histopathology of stapled haemorrhoidopexy specimens. PMID- 12780899 TI - Surgery for carcinoid tumours of the lower gastrointestinal tract. AB - INTRODUCTION: Carcinoid tumours are rare neoplasms that originate from neuroendocrine cells of the primitive gastrointestinal tract. Mid- and hind-gut tumours comprise the majority of these rare tumours. With many recent advances in medical treatment the role and importance of surgery and the surgeon needs to be assessed. METHOD: A Medline, Pubmed and Embase databases search was undertaken. All relevant articles were cross-referenced. RESULTS AND CONCLUSIONS: Incidental findings of carcinoid tumours should be treated at initial surgery whilst elective surgery and further management should be undertaken in specialist centres by a multidisciplinary team. Asymptomatic patients have a better prognosis than those with symptoms. In advanced cases surgery combined with chemotherapy and liver resection is appropriate. The outlook for the majority of cases is good. PMID- 12780900 TI - Management of soft tissue sarcomas of the abdomen and pelvis. AB - OBJECTIVE: Soft tissue sarcomas of the abdomen or pelvis are rare tumours that usually arise in the retroperitoneum but may also arise from the gastrointestinal tract or other abdominal organs. The aim of this study was to assess patient outcome following treatment of these tumours at a tertiary referral centre. PATIENTS AND METHODS: Data on all patients with soft tissue tumours treated under the care of one surgeon between January 1990 and December 2000 was reviewed. RESULTS: Over the 11-year period 61 patients underwent 79 operations for a soft tissue sarcoma of the abdomen or pelvis. Macroscopic clearance of tumour at operation was achieved in 45 (74%) of the 61 patients, for primary tumours this was 97%, recurrent tumours 50% and metastases 33%. The median survival for those undergoing complete resection was 50 months (range 3-238 months, median follow-up 33 months). This compared with a median survival of only 6 months range (1-127 months) for those that had incomplete resection (P = 0.001). The 5-year survival for patients that had complete resection was 61.5%, however, over one-third of the 5-year survivors have since recurred. CONCLUSION: This study confirms that most primary soft tissue sarcomas of the abdomen or pelvis are resectable and the majority of patients will experience a long disease free interval following resection. PMID- 12780901 TI - A comparison of the simultaneous application of sclerotherapy and rubber band ligation, with sclerotherapy and rubber band ligation applied separately, for the treatment of haemorrhoids: a prospective randomized trial. AB - OBJECTIVE: To compare simultaneous application of sclerotherapy and rubber band ligation, with sclerotherapy and rubber band ligation applied separately for the treatment of 2nd degree haemorrhoids. PATIENTS AND METHODS: Between 1993 and 1996, 255 patients that suffered from 2nd degree haemorrhoids were divided into 3 groups of 85 patients, each to receive either simultaneous sclerotherapy for smaller and rubber band ligation for larger piles (SCL/RBL) in one session, or sclerotherapy (SCL), or rubber band ligation (RBL), respectively. After a period of 4 years all patients were examined and their symptoms were recorded. RESULTS: The patients of the SCL group developed significantly fewer complications after treatment compared to the other two methods (P < 0.001), which did not differ from each other. After the SCL/RBL treatment, significantly more patients were symptom free (46%) than after SCL (8%), P < 0.001. There was no significant difference between the SCL/RBL (46%) and the RBL (31%) groups (P = 0.217), although the combined treatment seemed to be more effective than rubber band ligation. Only 10% of the patients of the SCL/RBL group needed additional sessions 6-24 months after the initial treatment compared to 30% of the patients of the SCL group (P = 0.001). However, there was no significant difference between SCL/RBL and RBL (17%) groups (P = 0.151). CONCLUSION: The combination of sclerotherapy and rubber band ligation for treatment of 2nd degree haemorrhoids is significantly more efficient than sclerotherapy on its own. PMID- 12780902 TI - Colorectal cancer in the south of Israel; comparison of the clinical characteristics and survival between two periods, 1981-2 and 1986-7. AB - BACKGROUND: Colorectal cancer has the highest incidence of all malignant tumours in men and women in Israel. The public was introduced to the concept of screening in the early 1980s, and full programs began in 1983. OBJECTIVES: The study compares patients with colorectal cancer (CRC) who were diagnosed before screening and five years later, for stage differences and survival. METHODS: All patients with CRC who were diagnosed from 1981 to 1982 (period I) and 1986-87 (period II) at Soroka Medical Centre and who were residents of the Negev area were eligible for the study. Data were collected retrospectively from the Israel Cancer Registry and patient files and endoscopic, histological and surgical reports from the Departments of Pathology, Gastroenterology, Oncology and Surgery at Soroka Medical Centre. RESULTS: The study groups included 207 patients with CRC, 85-from period I and 122-from period II. The mean annual incidence of CRC for these 2 periods was 18 and 24.8 cases per 100, 000, respectively, for an increase of 37% (P < 0.001). Five-year-survival in period I was 53.3% and in period II, 39.6% (P = 0.025). Survival did not differ by origin or gender. Dukes' stage, well and moderately differentiated tumours, and left-sided tumours were associated with better prognosis (P = 0.03). Forty-two percent of the cases were less than 64-year-old-at diagnosis, compared with 27% of 2069 Israeli patients in whom CRC was diagnosed in 1991 (P < 0.001). CONCLUSIONS: Before a nation wide decision is made with regard to public screening for CRC with fecal occult blood tests or sigmoidoscopy, the available data should be thoroughly evaluated, and further in-depth investigations performed to determine trends in diagnosis and prognosis. PMID- 12780903 TI - Iron-deficiency anaemia and delay in the diagnosis of colorectal cancer. AB - AIMS: Iron-deficiency anaemia (IDA) is a recognized complication of colorectal cancer (CRC) especially with right-sided tumours, and failure to investigate the anaemia in older patients may lead to a delay in diagnosis. The aims of this study were to establish the proportion of patients with CRC shown to have an IDA for more than six months before diagnosis and to establish the proportion of patients with IDA who subsequently prove to have CRC. METHODS: All patients presenting with confirmed CRC in a health district (catchment population 280000) in the 4 years 1996-9 were identified from the pathology database after ethical approval. The criteria for IDA were haemoglobin (Hb) < 10.1 g/dl plus mean corpuscular volume < 78 fl and/or mean corpuscular Hb concentration < 32 g/dl. The haematology data-base serving the same population was searched for evidence of: 1. IDA at diagnosis of CRC; 2. IDA more than 6 months and more than one year before the diagnosis of CRC; 3. The number of haematology referrals per annum in women over 55 and men over 50 years of age meeting the criteria for IDA. RESULTS: Of 440 patients with colorectal cancer, 166 (38%) had IDA at diagnosis and of the latter 54 (12%) were known to have IDA for more than six months before diagnosis and 26 (6%) had IDA more than one year before diagnosis. IDA was more common in right sided tumours (65%) than in those arising in the left side of the colon and rectum (26%). The annual incidence of IDA in the sampled population was 1366 in the stated age group. CONCLUSION: The investigation of iron-deficiency anaemia in older patients is important but in order to detect 26 patients with colorectal cancer a year earlier, the investigation of approximately 5000 patients would be required--a detection rate of less than 1%. PMID- 12780904 TI - The variable effect on proliferation of a colon cancer cell line by the citrus fruit flavonoid Naringenin. AB - OBJECTIVE: Naringenin, a naturally occurring flavonoid found in citrus fruits, is known to have anticarcinogenic properties. We have examined the effect of Naringenin on cell proliferation of an HT-29 colon cancer cell line. METHODS: HT 29 colon cancer cells were cultured in 96-well tissue culture plates. Naringenin concentrations ranging from 0.02 to 2.85 mmol were added to the wells of the Test group. The Control group contained all the elements present in the Test group with the exception of Naringenin. Cell proliferation was measured by colourimetric assay using the 2% WST-1 cell proliferation kit. RESULTS: Significant inhibition of cell proliferation was observed in HT29 colon cancer cells exposed to Naringenin at doses greater than 0.71 mmol. CONCLUSIONS: These results suggest a potential role for citrus fruits as a source of chemoprotective agents for colon cancer. PMID- 12780905 TI - Double stapled ileal pouch anal anastomosis (DS-IPAA) for mucosal ulcerative colitis (MUC): is there a correlation between the tissue type in the circular stapler donuts and in follow-up biopsy? AB - PURPOSE: This study aims to assess the correlation between the tissue types found in the circular stapler donut at the time of initial double-stapled ileal pouch anal anastomosis (DS-IPAA) and during subsequent periodic routine random biopsy. Secondarily, we sought to assess the risk of dysplasia, carcinoma or mucosal ulcerative colitis (MUC) recurrence in the retained mucosa. METHODS: The pathology reports of 91 patients (48 males, 43 females) who were operated upon for MUC from September 1988 to June 1997 and were reviewed and had two follow up visits for biopsy. The histological features of the distal donuts and biopsies of retained mucosa obtained at yearly interval follow-up were assessed in order to determine the epithelial tissue type (columnar, transitional and squamous), inflammation, recurrence of MUC and presence of dysplasia or malignancy. RESULTS: Median age at surgery was 43 (range 15-71) years and duration of MUC was 9.6 (range 0.3-42) years prior to surgery. The anastomosis was performed at a median height of 1.0 (range 0-2.5) cm cephalad to the dentate line and biopsy follow-up was undertaken at median 34 (range 2-110) months after DS-IPAA. The distal donuts were analysed in all cases, as were 305 follow-up biopsies (median 3.4; range 1-7 per patient). Although columnar epithelium (CE) was found in 62 (68%) donuts, it was absent on follow-up biopsy in 16 (26%) of these patients. Conversely, although no CE was identified in 29 (32%) donuts, it was identified in 11 (38%) of these patients during follow-up biopsy. CE in the donut was a significant predictor of CE in subsequent biopsies (P = 0.0012). The histological features consistent with MUC were seen in the biopsies from the retained mucosa in 15 (16%) patients from 0.3 to 7.6 years after DS-IPAA. While eight (9%) patients exhibited dysplasia or adenocarcinoma in the excised colon or rectum, none of the patients had either dysplastic changes or carcinoma within the retained mucosal biopsies. CONCLUSION: The correlation between CE in the circular stapler donut and at follow-up biopsy was high. However since CE developed in some patients in whom no CE was present in the distal donuts, regardless of the epithelial tissue type finding at the time of DS-IPAA, periodic follow-up biopsy should be obtained. PMID- 12780906 TI - Local excision of rectal carcinoma not-exceeding the muscularis layer. AB - AIM: The aim of the study was to compare the results of treatment by local excision of two different clinical stages of the rectal cancer. MATERIAL AND METHODS: Fifty-eight patients with early rectal carcinoma were operated on during the last 26 years using different methods of local excision. The carcinomas were initially assessed as not-exceeding the muscularis layer of the rectal wall. The tumours, localized up to 12 cm from the anal margin, were removed by means of "parachute" excision (47 patients). In 6 patients, carcinoma localized in the central part of rectum, was excised by means of the Localio method. Transanal endoscopic microsurgery was applied in 5 cases of carcinoma localized on the depth of 5-20 cm from the anal margin. RESULTS: After local excision the patients were divided into two groups: I, tumours of low degree of malignancy, not exceeding submucosal layer (26 patients); II, tumours of low or median degree of malignancy with infiltration of muscularis layer (32 patients). There was a significant difference in cancer relapses between groups I and II. One patient in group I and 9 in group II developed local recurrences (P < 0.05) and 5 patients in group II had neoplastic dissemination (15.6%). CONCLUSIONS: Best results were obtained in patients with carcinoma not exceeding submucosal membrane. In cases of rectal muscular layer infiltrations, the risk of carcinoma relapses was markedly higher. The use of transanal endoscopic microsurgery has permitted removal of tumours from the upper rectum. PMID- 12780907 TI - Micrometastasis in regional lymph nodes of extirpated colorectal carcinoma: immunohistochemical study using anti-cytokeratin antibodies AE1/AE3. AB - OBJECTIVE: The main objectives of this study were to identify, by immunohistochemistry, possible micrometastasis in the regional lymph nodes previously considered free by conventional histopathological examination, and to assess their influence on the survival of patients with colorectal cancer that had been extirpated in a radical manner. PATIENTS AND METHODS: From 38 patients with Dukes B staging (Colorectal Carcinoma Stage II (T3 N0 M0 or T4 N0 M0)) colorectal carcinoma, 383 lymph nodes were studied in paraffin blocks that had previously been considered free by conventional histopathological examination. These were submitted to immunohistochemical study using AE1/AE3 anti-cytokeratin monoclonal antibodies to identify neoplastic epithelial cells. RESULTS: Seven lymph nodes (1.82%) in six patients (15.78%) contained micrometastasis. The survival of the patients with extirpated colorectal carcinoma staged as Dukes B who had lymph node metastasis was less than in the group of patients without micrometastasis, although these values were not statistically significant. CONCLUSION: This immunohistochemical method can be employed successfully in the detection of neoplastic cells in lymph nodes previously considered free. In this study, there was a trend towards lower survival in node-positive patients but this did not reach statistical significance. PMID- 12780908 TI - Does the need to self-digitate or the presence of a large or nonemptying rectocoele on proctography influence the outcome of transanal rectocoele repair? AB - INTRODUCTION: Transanal repair of anterior rectocoele is a well described technique with variable success rate. In our department we offer repair to patients who fit the following criteria; the need to self digitate (transvaginal or perineal); a large rectocoele; a nonemptying rectocoele. Using these selection criteria previous authors have shown excellent results. The aim of our study was to review our results using this selective approach and also to determine whether the need to self digitate, the presence of a large rectocoele and the degree of emptying could predict which patients would achieve a successful result. METHODS: Fifty-five patients underwent repair over a three-year period. The main presenting symptom was noted for each patient and also whether self-digitation was required in order to achieve successful evacuation. Dynamic evacuation proctography was performed on all patients. Size of rectocoele, percentage of paste expelled and other proctographic abnormalities were noted for each patient. Follow up was at 6 weeks and 6 months at which point patients were asked whether their symptoms had resolved, improved, remained the same or had worsened. RESULTS: Complete data were available for 48 of the patients (median age 52 years, IQR 43-63). The presenting complaint was constipation in 22 patients, obstructive defaecation in 15, incomplete evacuation in 5, postdefaecation soiling in 4 and dyspareunia in 2. Thirty-eight patients noted the need to self digitate, 10 did not. Proctography revealed a large rectocoele (> 4 cm) in 22 patients and a medium or small rectocoele (< 4 cm) in 26 patients. There was a rectocoele alone in 34 patients, in combination with perineal descent in 11 and with intussusception in 3. Median percentage of paste expelled was 70% (range 20 95). At 6 weeks postoperatively, 43 patients had complete resolution of their symptoms whilst 5 reported only some or no improvement. At 6 months, 37 patients sustained complete resolution of their symptoms and 11 did not. Pre-operative factors were compared for these two groups of patients. There was no difference in age (P > 0.05, Mann-Whitney U-test) between the two groups There was also no difference in size of rectocoele, degree of emptying, the presence of another proctographic abnormality and the need to self-digitate between the two groups (P > 0.05, Fisher's exact test). DISCUSSION: No factors were seen to distinguish between the successful and unsuccessful groups of patients following rectocoele repair, however, an overall success rate of 75% was achieved using our selection criteria. This figure is in keeping with reported success rates in the literature. PMID- 12780909 TI - Quality of life before and after proctocolectomy and IPAA in patients with ulcerative proctocolitis--a prospective study. AB - OBJECTIVE: The principal aim of the present investigation was to study the general QoL and disease-specific adjustment before, and one-year after, operation with an ileal pouch anal-anastomosis (IPAA) in a consecutive series of patients with ulcerative colitis (UC). PATIENTS AND METHOD: Eleven patients were medically treated and 31 patients underwent colectomy with ileostomy. The patients were interviewed before the IPAA operation and again one-year after closure of the covering loop ileostomy. General quality of life was estimated with a Swedish instrument (according to Kajandi, score range 16-96) and a VA-scale. The instrument for disease-specific adjustment contained specific questions with five factors (score range 34-204). Open questions about quality of life were used to identify areas of concern, which were not included in the other instruments. Postoperative function was described in terms of a functional score (score range 0-15). RESULTS: The results of the general quality of life assessments did not differ significantly when comparing pre-operative to postoperative values. Disease-specific adjustment showed that the medically treated patients had a score of 162; interquartile range (IQR) 145-176. The patients with ileostomy scored 164; IQR 141-180. Postoperatively there was a statistically significant improvement in both groups to 189 (IQR 172-199), an increase of 15%. The open questions revealed restrictions in daily life for both patient groups pre operatively with improvements after IPAA. The median grading of functional outcome according to the score system was 2. The most frequent problems were: patients who had occasional bowel movements at night, 40%; intermittent perianal soreness, 51%; antidiarrhoeal medication use, 61%. CONCLUSION: General QoL did not change after IPAA. The bowel-specific adjustment in most factors improved in the majority of patients after IPAA. However, half of the IPAA patients had occasional bowel movements at night perianal soreness, and used antidiarrhoeal medication. PMID- 12780910 TI - Nationwide decline in annual numbers of abdomino-perineal resections: effect of a successful national trial? AB - OBJECTIVE: Large national trials may influence surgical practice. In this study the relation between the successful national randomized trial on the management of rectal cancer (the Dutch TME trial) and national ratio of abdomino-perineal resection to low anterior resection and anastomosis was analysed. PATIENTS AND METHODS: In the study period, 1994-99, 15978 patients underwent either abdomino perineal resection (n = 2575) or low anterior resection and anastomosis (n = 13403). The Dutch TME trial started in 1996 and a total of 1530 patients were included by 83 hospitals and 82.1% of these patients were treated from 1997 to 1999. Teaching sessions, tutor assisted surgery and quality control formed an integral and important part of the TME trial. RESULTS: Ratio of abdomino-perineal resection vs. low anterior resection was compared between period I (1994-96) and period II (1997-99). The ratio decreased from 0.19 to 0.13 between period I and II (95% CI, -0.08 to -0.04, P < 0.001). In hospital mortality rate did not change between period I and II (3.5 vs. 3.7, 95% CI, -0.08 to 0.03, P=0.385). CONCLUSION: Significant changes in surgical attitude may accompany successful national randomized trials in which investigated surgical procedures are specified, taught, and controlled. The APR ratio declined by 32% in the Netherlands during and following the Dutch TME trial, without a rise in hospital mortality rate for rectal resections. PMID- 12780911 TI - Colorectal transport in normal defaecation. AB - OBJECTIVE: Colorectal luminal transport during defaecation can be assessed by means of scintigraphy. However, normal values remain to be established and inter- and intra-subjective variation is unknown. The aim of the study was to describe colorectal transport during normal defaecation by means of a new method for colorectal scintigraphy and to determine inter- and intrasubjective variation. METHODS: Thirty healthy volunteers (18 men and 12 women, age 22-46 years; median 31) took 2 MBq 111In on two consecutive days (days 0 and 1). On days 1 and 2 an abdominal scintigraphy was done before and after defaecation. Changes in colorectal contents were computed in two ways: firstly, assuming ordered evacuation, the extent of colorectal emptying was computed; secondly, antegrade and retrograde transport within the colorectum was computed for each colorectal segment. RESULTS: Median colorectal emptying during normal defaecation was 99% of the rectosigmoid (range: 60% of the rectosigmoid to complete emptying of the rectosigmoid, descending colon, transverse colon and 19% of the caecum/ascending colon). There was no difference between men (median 99% of the rectosigmoid) and women (98% of the rectosigmoid). Day to day variation was large (dispersion=52%) with a trend towards larger defaecation on day 2 than on day 1 (P = 0.09). Large antegrade and retrograde movements of colorectal contents were observed within the colorectum during defaecation. Antegrade transport involved the rectosigmoid (median 99%), the descending colon (median 53%), the transverse colon (median 46%), and the caecum/ascending colon (median 11%). Retrograde transport was mainly from the transverse colon and the descending colon. Colorectal emptying was significantly correlated to the subjective description (P < 0.01) but not to total gastrointestinal or segmental colorectal transit times determined by means of radioopaque markers. CONCLUSIONS: Colorectal scintigraphy before and after defaecation provides detailed information about colorectal emptying and intracolonic antegrade or retrograde transport during defaecation. Inter- and intraindividual variations are large and antegrade and retrograde transport within the colorectum is common. PMID- 12780912 TI - Use and influence of the internet on patients undergoing ileoanal pouch surgery. PMID- 12780913 TI - Re: A novel trans-Denonvillier's fascial abdomino-perineal approach to a "buried" rectal stump when reconstructing a Hartmann's procedure. PMID- 12780914 TI - Re: Kelly: Cancer Services Collaborative Project. PMID- 12780915 TI - Residual staples as a possible cause of recurrent rectal bleeding after stapled haemorrhoidectomy. PMID- 12780920 TI - Colorectal nurse specialists. PMID- 12780921 TI - Management of colorectal liver metastases. AB - Hepatic metastases occur in 60% of patients following resection for colorectal cancer. Liver resection is the only curative option, with one third of resected patients alive at five years. In those developing recurrence in the liver following resection, further liver surgery may be curative, with similar 5 years survival rates of about 30%. Until recently surgery was feasible in only 15-25% of patients with colorectal liver metastases. New strategies, such as downstaging chemotherapy, portal vein embolization and two-stage hepatectomy, may increase the resectability rate by 15%. Earlier detection of liver metastases would increase resectability, although good follow-up trials are lacking. Once suspected, colorectal liver metastases are staged by spiral CT, CT portography and MRI, which have similar overall accuracies. Mortality following liver resection is less than 5% in major centres, with a morbidity rate of 20% to 50%. Prognostic scoring systems can be used to predict the likely cure rate with resection. Pulmonary metastases occur in 10-25% of patients with resected colorectal cancer, but are limited to the lung in only 2% of cases. In these selected cases surgery provides long-term survival in 20-40%, and repeat lung resection has shown similar rates. For patients with unresectable disease, chemotherapy and ablation techniques have been demonstrated to prolong survival, although chemotherapy alone has been shown to improve quality of life. PMID- 12780923 TI - Stapled anoplasty for haemorrhoids: a comparison of ambulatory vs. in-patient procedures. AB - OBJECTIVE: Haemorrhoids are commonly seen in colorectal practice. Stapled anoplasty is a novel approach to the treatment of this condition and is usually performed as an in-patient procedure. The aim of this study was to investigate the suitability of this technique for ambulatory surgery. PATIENTS AND METHODS: Fifty consecutive patients undergoing stapled anoplasty under general anaesthesia as day cases (DC) (mean age 41 years; 27 females) by a single consultant surgeon over a 12-month period were compared with 50 consecutive patients undergoing the same procedure as in-patients (mean age 44 years; 25 females) (IP) during the same period. RESULTS: Eight DC patients (16%) were admitted overnight from the day surgery unit for urinary retention (3), pain (2), bleeding (2) and anaesthetic reasons (1). Three other DC patients were re-admitted after a mean period of 4 days with bleeding (2), one of which required surgical haemostasis, and a septic complication (1). Mean hospital stay for IP cases was 2.6 (range 1 9) days. Two IP cases were re-admitted after 4 and 11 days for bleeding and wound infection, respectively. At review 2-4 weeks after discharge, satisfaction in both groups was high. Minor staple-line strictures were seen in 1 DC and 2 IP cases but all were easily dilated digitally. Mean costs incurred were significantly less for day surgery patients. CONCLUSIONS: Stapled anoplasty is suitable for use in day-case surgery as it is a quick and relatively painless procedure. The advantages, particularly financial, support the technique for use in an ambulatory setting, preferably in the morning, and provided detailed patient advice is given. PMID- 12780922 TI - Formaldehyde application for haemorrhagic radiation-induced proctitis: a clinical and histological study. AB - OBJECTIVES: Haemorrhagic radiation-induced proctitis is a serious complication of radiotherapy of pelvic organs. In severe cases, massive haemorrhage may necessitate hospitalization and repeated transfusions. Application of formaldehyde under direct vision is one of the most efficient treatments. The aim of this study was to evaluate the results of this treatment as well as the histological changes induced by formaldehyde on the rectal mucosa. MATERIAL AND METHODS: From January 1991 to September 2001, 13 patients who presented a haemorrhagic radiation-induced proctitis have been treated in our outpatient clinic with 4% formaldehyde cotton soaked applications. They were followed up to one year after the treatment. Endoscopic biopsies were performed before, immediately after the application, 1 month later, as well as at the one-year follow-up. RESULTS: In eight cases bleeding stopped after the first application. In two patients a second application was necessary to control the haemorrhage and in two other patients bleeding ceased definitively after the fourth application. Follow-up evaluation at 12 months showed no sign of acute proctitis or rebleeding. One asymptomatic patient had a mild stenosis of the rectum. Baseline biopsies showed signs of acute inflammation. Those performed after the application of formaldehyde showed fresh thromboses of the vessels of the mucosa. Biopsies at 1 month and 1 year showed only chronic changes secondary to the radiotherapy. CONCLUSION: Local application of 4% formaldehyde for the treatment of haemorrhagic radiation-induced proctitis gives good results, is well tolerated and easy to perform. Formaldehyde applied selectively causes thromboses of the bleeding vessels, without deep lesions or extended necrosis. PMID- 12780924 TI - A 5-cm colonic J pouch colo-anal reconstruction following anterior resection for low rectal cancer results in acceptable evacuation and continence in the long term. AB - BACKGROUND: Optimal treatment for low rectal cancer is total mesorectal excision, with most patients suitable for low colo-rectal or colo-anal anastomosis. A colon pouch has early functional benefits, although long-term function, especially evacuation, might mitigate against its routine use. The aim of this study was to assess evacuation and continence in patients with a colon pouch, and to examine the impact of possible risk factors. METHODS: In 1998, all 102 surviving patients with a colon pouch, whose stoma had been closed for more than one year, were sent a postal questionnaire. A composite incontinence score was calculated from questions on urgency, use of a pad, incontinence of gas, liquid or faeces; and a composite evacuation score from questions on medication taken to evacuate, straining, the need and number of times returned to evacuate. RESULTS: The response rate was 90% (50 M, 42 F), with a median age of 68 years (IQR 60-78) and median follow-up of 2.6 years (IQR 1.7-3.9). The anastomosis was 3 cm or less from the anus in 45/92 (49%), and incontinence scores were worse in this group (P = 0.001). There were significantly higher incontinence scores in females (P = 0.014). Age, preoperative radiotherapy, part of colon used for anastomosis, postoperative leak and length of follow-up had no demonstrable effect on either score. CONCLUSION: Gender and anastomotic height were the only variables which influenced incontinence. Ninety percent of patients reported that their bowel function did not affect their overall wellbeing, and none would have preferred to have a stoma. PMID- 12780925 TI - Patients with familial adenomatous polyposis experience better bowel function and quality of life after ileorectal anastomosis than after ileoanal pouch. AB - OBJECTIVE: To evaluate the quality of life with emphasis on bowel function in patients undergoing either total colectomy with ileorectal anastomosis (IRA) or restorative proctocolectomy with ileal pouch-anal anastomosis (IPAA) for familial adenomatous polyposis (FAP). PATIENTS AND METHODS: All 151 patients operated on in our department between 1971 and 2000 were analysed retrospectively. Since 42 patients had to undergo reoperations, a total of 194 bowel procedures were performed. The final operative breakdown was: 48 IRA, 62 IPAA, 33 total proctocolectomies and 8 other procedures. By the end of the period under consideration, 40 patients had died, 28 had been lost to follow-up, 83 patients were still alive and of these 76 were analysed. Continence function and quality of life with emphasis on overall bowel function (Wexner (WS)-, Jostarndt (JS)-, Oresland (OS)- and Pemberton (PS)-scores) were recorded in 59 patients (22 IRA and 37 IAP). RESULTS: Continence and bowel function were significantly better after IRA than after IPAA (mean scores: WS, 1.1 vs 5.4, P=0.001; JS, 32.5 vs 24.7, P=0.0001; OS, 2.9 vs 7.9, P=0.0001), as was quality of life (PS, 25.6 vs 19.6, P=0.001). The mean values for each single item of all 4 scores were consistently better after IRA. Neither gender, age nor type of pouch-anal anastomosis (stapled vs hand-sewn) had a significant influence on outcome. IPAA patients with uneventful follow-up (n = 27) had better mean values for all 4 scores (WS, 4.2 vs 7.3; JS, 26.1 vs 23.1; PS, 20.4 vs 17.7; OS, 6.6 vs 9.9) than those with postoperative complications (n = 10; 27%), but statistical significance was reached only in the OS (P = 0.026). No such difference was seen after IRA (14% complication rate). CONCLUSION: The significantly better continence after IRA resulting in superior patient comfort and quality of life must be balanced against the oncological disadvantage of this procedure. On the basis of large patient population studies, genotype-phenotype correlations with respect to the risk for rectal cancer may need to be taken into account. Postoperative complications may have a lasting negative effect on outcome after IPAA. PMID- 12780926 TI - A novel indicator for surveillance colonoscopy following colorectal cancer resection. AB - INTRODUCTION: Current surveillance for recurrent intraluminal or metachronous colorectal cancer following resection is largely undertaken by colonoscopic examination of the remaining colon. The burden on colonoscopic services is high and the procedure is expensive. Immunological faecal occult blood testing (FOBT) is a sensitive and specific test for detecting colorectal cancer, and may fine tune the need for timely surveillance colonoscopy. METHODS: Consecutive patients due for surveillance colonoscopy following colonic resection for cancer were prospectively studied. Each patient had a single faecal sample obtained at per rectal examination on a gloved examining finger. This was subjected to immunological FOBT in the clinic, and patients were categorized as FOBT positive or negative, according to the result. Colonoscopy as well as ultrasound or CT of the liver were performed within eight weeks of FOBT. RESULTS: Six hundred and eleven patients had both FOBT and colonoscopy. Fifty-nine (13.6%) were categorized as FOBT-positive. Of these, nine had biopsy-proven recurrent or metachronous cancer, 12 patients had one, or more adenomatous polyps, one patient had radiation proctitis and two patients had pan-colonic mucositis following chemotherapy. In the remaining 552 FOBT-negative patients, no cancers were found. Thirty-eight patients had polyps that were removed. The sensitivity and specificity for detecting cancer by immunological FOBT was 100% sensitivity for detecting adenomatous polyps was 24% but specificity was 93%. CONCLUSION: The immunological faecal occult blood test provides sensitive detection of metachronous and recurrent cancer in postoperative surveillance. Routine application may be used to reduce the frequency of colonoscopic surveillance, as a negative FOBT may be taken as a sign that colonoscopy may be deferred safely. PMID- 12780927 TI - A prospective audit of stomas--analysis of risk factors and complications and their management. AB - AIM: To prospectively audit stomas and to determine the nature and rate of complications and their relationship with various risk factors and their management. MATERIALS AND METHODS: The study was performed prospectively on 97 consecutive patients who had stomas formed between January 2000 to August 2000. Patients were followed up for one year. Risk factors including age, body mass index (BMI), preoperative siting, contour of the abdominal wall, smoking, grade of operating surgeon, emergency or elective procedure, diabetes, type of stoma and suture material used were noted. The type of surgery, and indications for surgery were also recorded. The complications were documented by two qualified stoma nurses and a photographic record taken. Statistical analysis comprising both univariate and multivariate methods, was performed by SPSS 10. RESULTS: The mean age was 65 years (standard deviation 16.01, range 16-99) and mean body mass index was 24.5 (standard deviation 4.66, range 15-37). Forty-nine of 97 (50.5%) stomas developed one or more complications. Twenty-three patients experienced retraction, 18 had stomas sited in a skin crease, 16 had early and 12 had late skin excoriation, 12 had detachments and a further 12 had parastomal hernia. Eleven further stoma complications were noted including prolapse, necrosis, ischaemia and sloughing. None of the risk factors achieved statistical significance when analysed against the overall complication rate. However, when the risk factors were analysed against individual complications using univariate logistic regression, a high body mass index was associated with more retractions (P = 0.003), early skin excoriation (P = 0.036) and poor siting (stoma in crease) occurred more commonly in emergencies (P = 0.022). Diabetes was associated with late skin excoriation (P = 0.02). Multivariate logistic regressions confirmed an independent association of body mass index, diabetes and emergency surgery with complications. Forty-five of 49 patients who had complications needed some conservative management such as a convexity appliance. Four patients needed refashioning. CONCLUSION: Body mass index, diabetes and emergency surgery were the significant risk factors identified in our study. Overall complications compare favourably with other series. We found that preoperative siting by stoma nurses and the grade of operating surgeon did not affect the outcome. PMID- 12780928 TI - Rectal flap advancement--a simple and effective approach for the treatment of rectourethral fistula. AB - OBJECTIVE: Rectourethral fistula is a rare complication of prostatic surgery and other pelvic procedures. We report our experience of surgical repair of using a rectal advancement flap. PATIENTS: Three patients with rectourethral fistula following prostatic surgery were treated. Two patients had an anterior partial thickness of rectal flap advancement via a trans-anal approach without urinary or faecal diversion. In one patient a rectal flap repair was performed through a posterior transsphincteric approach following urinary and faecal diversion. RESULTS: No significant postoperative complications occurred. Healing was successful in each patient and faecal and urinary continence was normal. CONCLUSION: Transanal rectal advancement flap is a simple and effective technique for the treatment of a rectourethral fistula with no need for urinary or faecal diversion. PMID- 12780929 TI - Health related quality of life in Crohn's proctocolitis does not differ from a general population when in remission. AB - OBJECTIVE: All treatment in Crohn's disease, although palliative, aims at restoring full health. The objective of this study was to compare health-related quality of life and psychosocial conditions in patients with Crohn's proctocolitis with a general population. PATIENTS AND METHODS: One hundred and twenty-seven patients with Crohn's proctocolitis (median age 44 years, 44.1% men) were compared with 266 controls (median age 45 years, 50.7% men). A questionnaire consisting of the Short Form-36 (SF-36), the Psychological General Well-Being Index (PGWB) and a visual analogue scale (VAS) evaluating general health as well as questions regarding psychosocial conditions was used. Disease activity was evaluated by Best's modification of the classical Crohn's Disease Activity Index. RESULTS: Patients in remission had a health related quality of life similar to controls according to the SF-36 apart from general health where scores were lower (P < 0.01). Patients with active disease scored lower in all aspects of the SF-36 (P < 0.001 or P < 0.0001) as well as the PGWB (P < 0.0001). In a model for multiple regression including age, gender, concomitant small bowel disease, permanent stoma, previous colonic surgery, disease activity, duration, and aggressiveness, disease activity was the only variable negatively predicting all 8 domains of the SF-36 in the patient group (P < 0.001). The mean annual sick leave for patients and controls were 33.9 and 9.5 days (P < 0.0001), respectively. Sixty-eight percent of the patients and 78.4% of the controls (P = 0.04) were married or cohabited, 67.7% and 78.0% (P = 0.04), respectively, had children. CONCLUSION: The health related quality of life for patients with Crohn's proctocolitis in remission does not differ from the general population. The disease has, however, a negative impact on parenthood, family life and professional performance. PMID- 12780930 TI - Creation of a treatment protocol for nurse-led management of anal fissure. AB - OBJECTIVE: To create a protocol for colorectal nurse-led management of patients with idiopathic anal fissure. PATIENTS AND METHODS: The outcome of 135 patients with anal fissure presenting over a 3-year period to one (half time equivalent) Colorectal Surgeon in a District General Hospital, in whom initial management was centred on the use of 0.2% glyceryl trinitrate (GTN) was audited. Patients unavailable for complete follow-up, those with fissures of a specific aetiology and those in whom GTN was contra-indicated were excluded. 44% of patients were initially seen by a suitably trained and supported Nurse Practitioner (NP). RESULTS: Success rates for fissure healing with 0.2% GTN were similar to those reported in other studies. There were no differences in outcome between those patients managed by the NP and those managed by the Consultant/Specialist Registrar. As a result of the audit, a treatment protocol for use by Nurse Practitioners (as well as surgical staff), a patient information leaflet, and policy documents relating to NP management of anal fissures have been developed. CONCLUSION: It has been possible to construct a treatment protocol allowing rational management of patients with anal fissure by suitably trained and supported nurse specialists within their own clinic. This may be adapted to encompass future treatment developments. PMID- 12780931 TI - Delayed sphincter repair for obstetric ruptures: analysis of failure. AB - OBJECTIVE: The aim of this study was to examine the clinical results after anterior anal sphincter repair in patients with obstetric trauma and to evaluate possible risk factors for poor outcome. PATIENTS AND METHODS: In years 1990-99 anterior anal sphincter repair for anal incontinence due to obstetric trauma was performed in 39 patients at Helsinki University Central Hospital. Clinical examination with Parks' classification and patients' questionnaire with endoanal ultrasound (EAUS) were done before and after surgery. Pudendal nerve terminal motor latency (PNTML) was measured postoperatively. The median follow-up time after the operation was 22 months (range 2-99). RESULTS: The follow-up results of the patients' questionnaire for 12 patients (31%) were good, for 15 patients (38%) acceptable and for 12 patients (31%) poor. Postoperative EAUS showed sphincter overlap in 28 (72%) patients but a defect was still found in 11 (28%) patients. A defect found on postoperative EAUS correlated with poor clinical result according to Parks' (R = 0.8, P < 0.01) and patients' questionnaire results (R = 0.7, P < 0.01). Patients with poor clinical results (Parks III/IV) were statistically significantly older (median 63 years, range 26-73) than those with favourable results (Parks I/II) (median 45 years, range 27-79) (P < 0.05). Further, the duration of incontinence symptoms correlated with poor functional results (R=0.4, P < 0.05). CONCLUSION: After obstetric trauma anterior anal repair gives acceptable short-term clinical results. EAUS investigation is easy and harmless to perform and should be used pre- and post-operatively. Advanced age, pre-operative signs of perineal descent, long-lasting severe incontinence symptoms and a persistent defect on postoperative EAUS seem to be related to poor clinical result. PMID- 12780932 TI - Re: Acute rectal obstruction after PPH stapled haemorrhoidectomy. PMID- 12780933 TI - European association of coloproctology annual meeting, Erlangen, Germany 12-14 september 2002. PMID- 12780936 TI - Comparative quantification of health risks conceptual framework and methodological issues. AB - Reliable and comparable analysis of risks to health is key for preventing disease and injury. Causal attribution of morbidity and mortality to risk factors has traditionally been conducted in the context of methodological traditions of individual risk factors, often in a limited number of settings, restricting comparability.In this paper, we discuss the conceptual and methodological issues for quantifying the population health effects of individual or groups of risk factors in various levels of causality using knowledge from different scientific disciplines. The issues include: comparing the burden of disease due to the observed exposure distribution in a population with the burden from a hypothetical distribution or series of distributions, rather than a single reference level such as non-exposed; considering the multiple stages in the causal network of interactions among risk factor(s) and disease outcome to allow making inferences about some combinations of risk factors for which epidemiological studies have not been conducted, including the joint effects of multiple risk factors; calculating the health loss due to risk factor(s) as a time-indexed "stream" of disease burden due to a time-indexed "stream" of exposure, including consideration of discounting; and the sources of uncertainty. PMID- 12780937 TI - Venous pseudo-aneurysm as a late complication of short-term central venous catheterisation. AB - Complications following central venous catheterisation abound in the medical literature. This reflects the under-estimated potential hazards associated with this procedure as well as an increase in the number of such procedures performed in high-risk patients. We report on a case where a venous pseudo-aneurysm developed four months after the removal of a short-term central venous catheter in a moderately high-risk patient. PMID- 12780938 TI - Blood lead concentrations in 1-3 year old Lebanese children: a cross-sectional study. AB - BACKGROUND: Childhood lead poisoning has not made the list of national public health priorities in Lebanon. This study aims at identifying the prevalence and risk factors for elevated blood lead concentrations (B-Pb >or= 100 microg/L) among 1-3 year old children. It also examines the need for universal blood lead screening. METHODS: This is a cross-sectional study of 281 well children, presenting to the pediatric ambulatory services at the American University of Beirut Medical Center in 1997-98. Blood was drawn on participating children for lead analysis and a structured questionnaire was introduced to mothers asking about social, demographic, and residence characteristics, as well as potential risk factors for lead exposure. Children with B-Pb >or= 100 microg/L were compared to those with B-Pb < 100 microg/L. RESULTS: Mean B-Pb was 66.0 microg/L (median 60.0; range 10-160; standard deviation 26.3) with 39 (14%) children with B-Pb >or= 100 microg/L. Logistic regression analysis showed that elevated B-Pb was associated with paternal manual jobs (odds ratio [OR]: 4.74), residence being located in high traffic areas (OR: 4.59), summer season (OR: 4.39), using hot tap water for cooking (OR: 3.96), exposure to kohl (OR: 2.40), and living in older buildings (OR: 2.01). CONCLUSION: Lead screening should be offered to high-risk children. With the recent ban of leaded gasoline in Lebanon, emphasis should shift to other sources of exposure in children. PMID- 12780939 TI - Finite element analysis of hepatic radiofrequency ablation probes using temperature-dependent electrical conductivity. AB - BACKGROUND: Few finite element models (FEM) have been developed to describe the electric field, specific absorption rate (SAR), and the temperature distribution surrounding hepatic radiofrequency ablation probes. To date, a coupled finite element model that accounts for the temperature-dependent electrical conductivity changes has not been developed for ablation type devices. While it is widely acknowledged that accounting for temperature dependent phenomena may affect the outcome of these models, the effect has not been assessed. METHODS: The results of four finite element models are compared: constant electrical conductivity without tissue perfusion, temperature-dependent conductivity without tissue perfusion, constant electrical conductivity with tissue perfusion, and temperature-dependent conductivity with tissue perfusion. RESULTS: The data demonstrate that significant errors are generated when constant electrical conductivity is assumed in coupled electrical-heat transfer problems that operate at high temperatures. These errors appear to be closely related to the temperature at which the ablation device operates and not to the amount of power applied by the device or the state of tissue perfusion. CONCLUSION: Accounting for temperature-dependent phenomena may be critically important in the safe operation of radiofrequency ablation device that operate near 100 degrees C. PMID- 12780940 TI - Sandwich hybridisation assay for quantitative detection of yeast RNAs in crude cell lysates. AB - BACKGROUND: A rapid microtiter plate based sandwich hybridization assay was developed for detection and quantification of single RNA species using magnetic beads. Following solution hybridization target RNA molecules were collected by biotin-streptavidin affinity binding and detected by fluorescence signal generated by alkaline phosphatase. The 18S rRNA and SUC2 mRNA of Saccharomyces cerevisiae were used as model RNA target molecules. RESULTS: The sensitivity of the assay was approximately 1.2 x 109 (2 fmol) molecules of target RNA. The developed method was feasible with crude cell lysates of S. cerevisiae carlsbergensis and was evaluated by measuring the levels of 18S rRNA during cell growth and SUC2 mRNA under repressive and inductive conditions. The 18S rRNA expression level followed the changes in the specific growth rate. SUC2 mRNA levels were in good correlation with the measured invertase enzyme activities. CONCLUSIONS: The here presented sandwich hybridisation method was succefully applied for monitoring the amounts of ribosomal RNA and mRNA with high expression level in shake flask cultivation conditions. Sandwich hybridisation method offers a fast and convenient tool for following single key RNA species of interest in the production conditions. PMID- 12780941 TI - Risk and resiliency factors in posttraumatic stress disorder. AB - BACKGROUND: Not everyone who experiences a trauma develops posttraumatic stress disorder (PTSD). The aim of this study was to determine the risk and resiliency factors for this disorder in a sample of people exposed to trauma. METHOD: Twenty five people who had developed PTSD following a trauma and 27 people who had not were asked to complete the Posttraumatic Stress Diagnostic Scale, the Coping Inventory for Stressful Situations, and the State-Trait Anxiety Inventory. In addition, they completed a questionnaire to provide information autobiographic and other information. ANALYSIS: Five variables that discriminated significantly between the two groups using chi-square analysis or t-tests were entered into a logistic regression equation as predictors, namely, being female, perceiving a threat to one's life, having a history of sexual abuse, talking to someone about the event, and the "intentionality" of the trauma. RESULTS: Only being female and perceiving a threat to one's life were significant predictors of PTSD. Taking base rates into account, 96.0% of participants with PTSD were correctly classified as having the disorder and 37.0% of participants without PTSD were correctly classified as not having the disorder, for an overall success rate of 65.4% CONCLUSIONS: Because women are more likely than men to develop PTSD, more preventive measures should be directed towards them. The same is true for trauma victims (of both sexes) who feel that their life was in danger PMID- 12780942 TI - A parsimonious explanation for intersecting perinatal mortality curves: understanding the effect of plurality and of parity. AB - BACKGROUND: Birth weight- and gestational age-specific perinatal mortality curves intersect when compared across categories of maternal smoking, plurality, race and other factors. No simple explanation exists for this paradoxical observation. METHODS: We used data on all live births, stillbirths and infant deaths in Canada (1991-1997) to compare perinatal mortality rates among singleton and twin births, and among singleton births to nulliparous and parous women. Birth weight- and gestational age-specific perinatal mortality rates were first calculated by dividing the number of perinatal deaths at any given birth weight or gestational age by the number of total births at that birth weight or gestational age (conventional calculation). Gestational age-specific perinatal mortality rates were also calculated using the number of fetuses at risk of perinatal death at any given gestational age. RESULTS: Conventional perinatal mortality rates among twin births were lower than those among singletons at lower birth weights and earlier gestation ages, while the reverse was true at higher birth weights and later gestational ages. When perinatal mortality rates were based on fetuses at risk, however, twin births had consistently higher mortality rates than singletons at all gestational ages. A similar pattern emerged in contrasts of gestational age-specific perinatal mortality among singleton births to nulliparous and parous women. Increases in gestational age-specific rates of growth-restriction with advancing gestational age presaged rising rates of gestational age-specific perinatal mortality in both contrasts. CONCLUSIONS: The proper conceptualization of perinatal risk eliminates the mortality crossover paradox and provides new insights into perinatal health issues. PMID- 12780943 TI - Patient-based studies of agreement between spirometers. PMID- 12780944 TI - Bronchial challenge: just do it! PMID- 12780945 TI - Comparison of a new desktop spirometer (Spirospec) with a laboratory spirometer in a respiratory out-patient clinic. AB - BACKGROUND: The performance of spirometers is often evaluated under ideal conditions with computer-generated waveforms or in vivo testing with healthy subjects. Real-life conditions are less ideal because of comorbidities, age of the subjects, and a variety of air flow limitations. Evaluation of new spirometry equipment can also be performed under these less favorable conditions. The Spirospec is a new desktop spirometer that is commercially available, but its accuracy has not been evaluated in a clinical setting. OBJECTIVE: Test the Spirospec with subjects with normal and pathologic pulmonary function. METHODS: A group of 45 patients (mean age 38.4 years, 27 male) booked for evaluation in the pulmonary function laboratory of a tertiary care university hospital were tested with both a Spirospec and a standard Jaeger Masterlab 4.0 spirometer, according to the guidelines of the American Thoracic Society. Three subgroups (normal spirometry, obstructive air flow limitation, and restrictive air flow limitation) of 15 consecutive subjects each underwent spirometry. RESULTS: Pulmonary function measurements from the Spirospec correlated closely (r = 0.95-0.99) with those from the Masterlab 4.0, showing good limits of agreement and differences between the 2 devices: forced vital capacity 0.03 L, forced expiratory volume in the first second (FEV(1)) -0.01 L, peak expiratory flow -0.41 L/s, peak inspiratory flow 0.43 L/s, forced expiratory flow at 50% of total lung capacity 0.13 L/s, and forced expiratory flow at 75% of total lung capacity 0.12 L/s. With the exception of forced vital capacity and FEV(1), these differences were statistically significant (p < 0.05). CONCLUSION: The Spirospec is comparable to the Masterlab 4.0, with high accuracy for FEV(1) and forced vital capacity and clinically acceptable differences in the measured flow variables. PMID- 12780946 TI - Clinical variables are poor selection criteria for the use of methacholine bronchoprovocation in symptomatic subjects. AB - OBJECTIVE: Airway hyperresponsiveness (AHR) is associated with persistent air flow limitation and accelerated FEV(1) decline. AHR can influence diagnosis, treatment, and prognosis. We assessed the value of pulmonary function variables, symptoms, and history as selection criteria for methacholine bronchoprovocation testing to detect AHR in symptomatic subjects. METHODS: Over a 4-year period we conducted a prospective study of consecutive subjects who underwent methacholine bronchoprovocation testing. Baseline pulmonary function testing (PFT) and a questionnaire were obtained prior to methacholine bronchoprovocation testing. PFT and symptom and history variables were assessed as AHR predictors in univariate and multiple logistic regression analyses for the whole group and for 4 different age groups. RESULTS: There were 530 subjects, with ages ranging from 5 to 87 years, and 232 (44%) were positive for methacholine AHR. AHR was more prevalent among subjects < or = 25 years old (59%) and > 65 years old (47%) than among the other age groups. PFT values, symptom, and history variables had different AHR predictive values among the different age groups. Symptom and history variables had no AHR predictive value among subjects < or = 25 or > 65 years old. CONCLUSIONS: Young and elderly symptomatic subjects are more likely to have methacholine AHR. None of the clinical variables we studied has significant predictive value for methacholine AHR across the age groups, so these variables are poor selection criteria for methacholine bronchoprovocation testing of symptomatic subjects. Given the high prevalence of AHR among these subjects, bronchoprovocation should be considered with all individuals who have respiratory symptoms of wheezing, cough, shortness of breath, or chest tightness. PMID- 12780947 TI - The role of transbronchial needle aspiration in the diagnosis of bronchogenic carcinoma. AB - BACKGROUND: For many years in the United States transbronchial needle aspiration (TBNA) has been used with flexible bronchoscopy to diagnosis bronchogenic carcinoma, but very few data are available from the United Kingdom. METHODS: All bronchoscopies performed for suspected bronchial carcinoma at Papworth Hospital, Cambridge, United Kingdom, over the last 3 years were reviewed retrospectively. Patients with peribronchial disease, as evidenced by submucosal infiltration or extrinsic compression on bronchoscopy, were selected for TBNA. Patients with computed tomography evidence of subcarinal lymphadenopathy were also included. In total we identified 78 patients: 67 with peribronchial disease and 21 with subcarinal lymphadenopathy. All 78 patients underwent TBNA, and in 8 of these TBNA was performed in 2 sites. RESULTS: Malignancy was confirmed in 66 of the 78 patients. TBNA was positive in 31/66 (47%) of the patients who had proven bronchogenic carcinoma. Additional staging information was obtained in 9/21 patients (42.8%) who underwent subcarinal lymph node aspiration. We also found that TBNA was diagnostic in 1 patient with tuberculosis and 1 with sarcoidosis. There was only 1 important TBNA complication, which was a small pneumothorax. CONCLUSION: In our preliminary experience with selected patients suspected to have bronchogenic carcinoma (based on peribronchial disease or subcarinal lymphadenopathy), we found TBNA a safe and useful tool. PMID- 12780948 TI - Chemotherapy-induced late acute respiratory distress syndrome following right pneumonectomy for bronchogenic carcinoma. AB - We report 2 patients who suffered late postoperative acute respiratory distress syndrome (ARDS) that was probably chemotherapy-induced. Both patients underwent neoadjuvant combination chemotherapy prior to right pneumonectomy for primary bronchogenic carcinoma, and then suffered ARDS in the remaining lung a few weeks after surgery. No evidence of infection or other specific ARDS etiologies could be found, whereas the bronchoalveolar lavage fluid cell differentiation and protein content suggested the permeability form of lung edema. Both patients had rapid clinical, functional, and radiologic improvement with high-dose corticosteroids. In the first patient the course was complicated by the development of a critical illness polyneuropathy with complete tetraplegia, but the patient recovered. The second patient died from septic shock 4 weeks after starting mechanical ventilation. The incidence of a chemotherapy-related ARDS in the remaining lung, occurring more than 4 weeks after extensive operations or after a pneumonectomy, is unknown. This kind of acute lung injury calls for particularly delicate treatments, the most potentially life-threatening complications being mainly associated with difficulties in ventilatory support and the high doses of corticosteroids required to rescue the remaining lung. PMID- 12780949 TI - Has oxygen administration delayed appropriate respiratory care? Fallacies regarding oxygen therapy. AB - Modern clinical use of supplemental oxygen supposes that: (1) exposure to F(IO)(2) < or = 60% is without adverse effects, (2) an individual at risk of developing arterial hypoxemia can be protected by administering high F(IO)(2), and (3) routine administration of supplemental oxygen is useful, harmless, and clinically indicated. There is now substantial evidence that none of those 3 suppositions are correct, and, on the contrary, supplemental oxygen is actually detrimental to many of the patients who receive it. Supplemental oxygen is much overused and its use should be limited to the few conditions and situations in which it is truly effective, useful, and non-detrimental. PMID- 12780950 TI - Survey on transfusion practices of pediatric intensivists. AB - OBJECTIVE: To describe the red blood cell transfusion practices of pediatric intensivists. DESIGN: Cross-sectional self-administered survey. SETTING: Pediatric intensive care units. PATIENTS: Academic pediatric intensivists. INTERVENTIONS: None. MEASUREMENTS AND RESULTS: Scenario-based survey among English- or French-speaking intensivists from Canada, France, Belgium, or Switzerland, working in tertiary-care pediatric intensive care units. Respondents were asked to report their decisions regarding transfusion practice with respect to four scenarios: cases of bronchiolitis, septic shock, trauma, and the postoperative care of a patient with Fallot's tetrad. The response rate was 71% (163 of 230). The overall baseline hemoglobin transfusion threshold that would have prompted intensivists to transfuse a patient ranged from 7 to 13 g/dL (70 130 g/L) within almost all scenarios. There was a significant difference between scenarios of the average baseline hemoglobin transfusion thresholds (p < .0001). A low Pao2, a high blood lactate concentration, a high Pediatric Risk of Mortality score, active gastric bleeding, emergency surgery, and age (2 wks) were important determinants of red blood cell transfusion, whereas none of the respondents' personal characteristics were. The average volume of packed red blood cells transfused in the four scenarios did not differ significantly. CONCLUSIONS: This survey documented a significant variation in transfusion practice patterns among pediatric critical care practitioners with respect to the threshold hemoglobin concentration for red blood cell transfusion. The volume of packed red blood cells given was not adjusted to the hemoglobin concentration. PMID- 12780951 TI - Association between the A/A genotype at the lymphotoxin-alpha+250 site and increased mortality in children with positive blood cultures. AB - INTRODUCTION: Tumor necrosis factor-alpha has been implicated in the hemodynamic manifestations of sepsis. Genetic polymorphisms located in the first intron of the lymphotoxin-alpha gene have been associated with increased secretion of tumor necrosis factor-alpha. We hypothesized that bacteremic children with the high secretor genotype, AA, have higher levels of tumor necrosis factor-alpha and a worse outcome. METHODS: Genotypic analysis was performed in children with bacteremia using polymerase chain reaction amplification and restriction enzyme digestion. The serum tumor necrosis factor-alpha levels were measured by enzyme linked immunosorbent assay technique. RESULTS: Genotypic frequencies at the lymphotoxin-alpha+250 site were 11 of 34 (0.32) AA, 16 of 34 (0.47) GA, and 7 of 34 (0.21) GG. Serum tumor necrosis factor-alpha levels were 324 +/- 124 pg/mL in bacteremic children with the AA genotype at the lymphotoxin-alpha+250 site compared with 92 +/- 59 pg/mL in bacteremic children with the AG genotype (p < .001) and 92 +/- 21 pg/mL in bacteremic children with the GG genotype (p < .001). Eight of 11 bacteremic children with the AA genotype died compared with 3 of 16 bacteremic children with the GA genotype (p < .001) and zero of seven bacteremic children with the GG genotype (p < .001). CONCLUSION: The AA genotype at the lymphotoxin-alpha+250 site is associated with higher serum tumor necrosis factor alpha levels and a higher mortality in children with bacteremia. PMID- 12780952 TI - Neurodevelopmental outcome of young pediatric intensive care survivors of serious brain injury. AB - OBJECTIVES: To determine the number of children with severe brain injury due to closed head injury or hypoxic-ischemic encephalopathy as a proportion of all admissions of children <3 yrs of age in the regional pediatric intensive care unit; to determine the outcome of these children at >6 mos postinjury; and to explore the relationship of outcome measures to predictors of outcome obtained within the first 24 hrs after brain injury. DESIGN: Prospective, descriptive outcome study of an inception cohort. RESULTS: Neonatal and Infant Follow-up Clinic, Glenrose Rehabilitation Hospital, Edmonton, Canada. PATIENTS: Of a cohort of 53 children of <3 yrs of age (4% of pediatric intensive care unit admissions, 1995-1998) admitted for severe acquired brain injury (Glasgow Coma Score, 80% have adverse outcome of death, disability, or mental or motor developmental scores below average. A Glasgow Outcome Scale of 5 overestimates good recovery. Universal registry and follow-up of these children are needed. PMID- 12780953 TI - Compared with specialist registrars, experienced staff nurses shorten the duration of weaning neonates from mechanical ventilation. AB - OBJECTIVE: To compare the overall performance of specially trained neonatal nurses acting autonomously, unsupervised, and without a protocol with specialist registrars when weaning neonates from mechanical ventilation. DESIGN: Prospective, randomized, controlled trial. SETTING: A single neonatal intensive care unit. PATIENTS: Neonates requiring conventional mechanical ventilation (n = 50). INTERVENTIONS: Infants on conventional ventilation were randomly assigned to receive either nurse-led (n = 25) or registrar-led (n = 23) weaning. A total of 48 infants completed the study (two infants in the registrar group were excluded when their parents withdrew consent). MEASUREMENTS AND MAIN RESULTS: The main outcome measure, median weaning time, was 1200 mins (95% confidence interval [CI], 621-1779 mins) in the nurse group and 3015 mins (95% CI, 2650-3380 mins) in the registrar group (p = .0458). The median time from treatment assignment to the first ventilator change was 60 mins (95% CI, 52-68 mins) in the nurse group and 120 mins (95% CI, 103-137 mins) in the registrar group (p = .35). On average, the nurses made ventilator changes every 4.5 hrs (95% CI, 2.9-6 hrs) and the registrars every 7.2 hrs (95% CI, 5.4-9 hrs; p = .003). The median number (range) of backward steps taken per infant was 0 (0-5 steps) in the nurse group and 1 (0 5 steps) in the registrar group (p = .019). CONCLUSIONS: The findings of this study suggest that additional domains of neonatal critical care could be reviewed for their potential transfer to appropriately prepared nurses. PMID- 12780954 TI - Successful local low-dose urokinase treatment of acquired thrombosis early after cardiothoracic surgery. AB - BACKGROUND AND OBJECTIVE: Recent surgery (1 yr, central-peripheral temperature gap and respiratory failure were major risk factors. CONCLUSIONS: The incidence of mortality is high in our setting and further research is needed to identify causes of preventable deaths. Children presenting with signs of hypoperfusion and respiratory failure had poor outcomes. This raises the concern that children may be presenting late, with advanced, severe illness to our pediatric emergency department. The data also suggest that identification of serious illness in children is possible with simple clinical signs and symptoms. PMID- 12780956 TI - Bleeding and coagulation changes during spinal fusion surgery: a comparison of neuromuscular and idiopathic scoliosis patients. AB - OBJECTIVE: Major blood loss is common during spinal fusion surgery. We have previously demonstrated that patients with neuromuscular scoliosis have more blood loss and greater transfusion requirement than those with idiopathic scoliosis. Our objective is to study the relationships between etiology of scoliosis, blood loss, and coagulation changes in children and adolescents undergoing spinal fusion surgery. DESIGN: Prospective, observational study. SETTING: University teaching hospital. PATIENTS: A total of 25 patients, 11 with neuromuscular and 14 with idiopathic scoliosis, undergoing spinal fusion surgery. INTERVENTIONS: Blood was obtained preoperatively, 2 and 4 hrs intraoperatively, and 2 and 24 hrs postoperatively for prothrombin time, partial thromboplastin time, thrombin time, platelet count, D-dimer, factor VII and VIII activity, thrombin-antithrombin III complex, and protein induced by vitamin K absence. Changes in coagulation over time were analyzed by repeated-measures analysis of variance. Comparisons between groups were made using independent t-tests. RESULTS: Neuromuscular scoliosis patients had significantly greater blood loss than idiopathic scoliosis patients (median blood loss, 78% of total blood volume; range, 25-127% vs. 20%, 2-82%; p < .001). Prothrombin time increased over time in both groups and was higher in the neuromuscular than the idiopathic group both preoperatively and postoperatively (p < .05). Factor VII activity decreased over time in both groups (p < .001) and was lower in the neuromuscular than the idiopathic group during surgery (p < .05). No changes in partial thromboplastin time, thrombin time, or factor VIII activity were observed. D-dimers were present in both groups by 4 hrs intraoperatively. Protein induced by vitamin K absence was not detected in any patient. CONCLUSIONS: Neuromuscular scoliosis patients have more blood loss during spinal fusion surgery than idiopathic scoliosis patients. The prolongation of prothrombin time and decrease in Factor VII activity suggest activation of the extrinsic coagulation pathway. Depletion of clotting factors during scoliosis surgery occurs to a greater extent in patients with underlying neuromuscular disease. PMID- 12780957 TI - Alternative arterial catheterization site using the ulnar artery in critically ill pediatric patients. AB - OBJECTIVE: To examine the patterns of usage and associated complications among radial, femoral, and ulnar artery catheters in the pediatric intensive care unit to ascertain the utility of the ulnar artery as an alternative vessel of peripheral cannulation. DESIGN: A 4-yr retrospective review of all admissions to the pediatric intensive care unit to identify all patients with ulnar, radial, or femoral arterial catheterization. SETTING: Pediatric intensive care unit of a tertiary care pediatric center. PATIENTS: A total of 124 critically ill pediatric patients (age, 0.4-36 mos) were identified. INTERVENTIONS: Ulnar and radial artery catheters were either 22 gauge or 24 gauge, femoral artery catheters were 2.5 Fr, 2.5 cm, or 3 Fr, 5 cm, in size. All catheters were percutaneously inserted. The ulnar artery was typically used when attempts at common peripheral sites were unsuccessful. MEASUREMENTS AND MAIN RESULTS: A total of 18 ulnar, 103 radial, and 69 femoral artery cannulations were identified. The ulnar artery catheter duration ranged from 1 to 19 days (mean, 4.2 +/- 4.2 days), radial artery duration ranged from 1 to 24 days (mean, 5.3 +/- 4.2 days), and femoral artery duration ranged from 1 to 21 days (mean, 6.9 +/- 4.5 days). Ischemia as the reason for catheter removal was 5.6%, 3.9%, and 8.7% for ulnar, radial, and femoral arterial catheters, respectively. The catheter-related infection rate was 0%, 0.97%, and 4.3% for ulnar, radial, and femoral arterial catheters, respectively. CONCLUSION: Based on the ischemic and infectious complication rates, the ulnar artery should be considered as a viable site for peripheral vessel cannulation in cases in which arterial catheterization is necessary and attempts at more common sites of catheterization are unsuccessful. PMID- 12780958 TI - Capillary and arterial blood gases in hemorrhagic shock: a comparative study. AB - OBJECTIVES: To determine whether capillary blood gas samples accurately reflect the information obtained from arterial blood gas samples in a porcine hemorrhagic shock model. DESIGN: Randomized prospective study. SETTING: Animal research laboratory at a children's hospital. SUBJECTS: Fifteen adult Yorkshire pigs. INTERVENTIONS: Tracheally intubated, ventilated, and anesthetized pigs were randomly assigned to either a control group (n = 10) or a hemorrhagic shock group (n = 5). After instrumentation, the hemorrhagic shock group underwent a 40-mL/kg hemorrhage over 2 hrs, while the control group did not. Animals were then monitored over a 3-hr period. MEASUREMENTS AND MAIN RESULTS: Simultaneous arterial blood gas and capillary blood gas samples were obtained from each animal by using standardized technique. Capillary blood gas pH accurately predicted arterial blood gas pH in both control and hemorrhagic shock groups (r = .94, p < .01, and r = .78, p < .01, respectively). Bland-Altman analysis revealed a mean difference between arterial blood gas pH and capillary blood gas pH of 0.016 (95% limit of agreement, +/-0.037) in the control group vs. a mean difference between arterial blood gas pH and capillary blood gas pH of 0.075 (95% limit of agreement, +/-0.142) in the hemorrhagic shock group. Capillary blood gas Pco2 accurately predicted arterial blood gas Pco2 in the control group but not in the hemorrhagic shock group (r = .80, p < .01, and r = .13, p = .41, respectively). By Bland-Altman analysis, the mean difference between arterial blood gas Pco2 and capillary blood gas Pco2 was 3.9, with a 95% limit of agreement of +/-3.61 in the control group. In the hemorrhagic shock group, the mean difference between arterial blood gas Pco2 and capillary blood gas Pco2 was 11.7 with a 95% limit of agreement of +/-38.8. Hemorrhagic shock group capillary blood gas samples significantly overestimated Pco2 (Delta = 11.7, p < .01) and underestimated pH (Delta = 0.075, p < .01) compared with arterial blood gas samples. CONCLUSIONS: Capillary blood gas samples are poor predictors of Pco2 and pH in porcine hemorrhagic shock. These findings should be taken into account when capillary blood gas samples are used in the monitoring and management of subjects diagnosed with shock. PMID- 12780959 TI - Aortoesophageal fistula in a 13-yr-old girl: complication after nasogastric tube placement in the setting of right-sided aortic arch. AB - OBJECTIVE: To report a case of aortoesophageal fistula in a 13-yr-old girl with a right aortic arch that occurred after nasogastric intubation after surgery for lumbar kyphosis. DESIGN: Case report. SETTING: Tertiary care pediatric intensive care unit. PATIENTS: One 13-yr-old girl who underwent operative repair of a 45 degree kyphosis at the level of the second to fourth lumbar vertebrae. MAIN RESULTS: On the eighth postoperative day after operative kyphosis repair and intraoperative placement of a nasogastric tube, sudden massive hematemesis developed and the patient died. Autopsy revealed esophageal ulceration with erosions. One of these had a fistulous tract connecting to the descending aorta. The aortoesophageal fistula was observed where the descending aorta indented the posterior wall of the esophagus, as the aorta crossed the midline from the right side to the left side of the body. CONCLUSION: In the setting of right-sided aortic arch and other abnormalities of the aortic arch, nasogastric intubation may result in aortoesophageal fistula, massive hemorrhage, and death. Right-sided aortic arch should be added to the list of conditions for which utmost caution during nasogastric intubation is warranted. PMID- 12780960 TI - Corticosteroids for the prevention of reintubation and postextubation stridor in pediatric patients: A meta-analysis. AB - OBJECTIVE: To determine whether corticosteroids are effective in preventing or treating postextubation stridor and in reducing the need for subsequent reintubation of the trachea in critically ill infants and children. DESIGN: Meta analysis of published randomized controlled trials. DATA SOURCES: References of each trial from a MEDLINE search were reviewed, and experts in the field were contacted. STUDY SELECTION: Any randomized controlled trial comparing the administration of corticosteroids with placebo on the prevalence of reintubation or postextubation stridor in infants or children receiving mechanical ventilation via an endotracheal tube in an intensive care unit. DATA EXTRACTION: Data extraction and methodologic quality assessment were assessed independently by two reviewers. DATA SYNTHESIS: Six controlled clinical trials met the criteria for inclusion; three trials pertain to neonates and three to children. Five trials examined the use of steroids for the prevention of reintubation (four of these evaluated postextubation stridor specifically); one trial examined the use of steroids to treat existing postextubation stridor in children. There was a nonsignificant trend toward a decreased rate of reintubation in all subjects when prophylactic steroids were used (n = 376, relative risk [RR] = 0.34, 95% confidence interval [CI] = 0.05-2.33). Prophylactic use of steroids reduced postextubation stridor in the pooled studies (n = 325, RR = 0.50, 95% CI = 0.28 0.88). In young children, there were significant reductions of postextubation stridor with preventive treatment (n = 216, RR = 0.53, 95% CI = 0.28-0.97), and a trend toward less stridor was observed in neonates (n = 109, RR = 0.42, 95% CI = 0.07-2.32). There was a nonsignificant trend toward a reduced reintubation rate when steroids were used to treat existing upper airway obstruction requiring reintubation (RR = 0.55, 95% CI = 0.17-1.78). Side effects were seldom reported and could not be evaluated. CONCLUSIONS: Prophylactic administration of dexamethasone before elective extubation reduces the prevalence of postextubation stridor in neonates and children and may reduce the rate of reintubation. PMID- 12780961 TI - Mortality risk factors of a pediatric population with fulminant hepatic failure undergoing orthotopic liver transplantation in a pediatric intensive care unit. AB - OBJECTIVES: To determine risk factors of mortality in the preoperative, perioperative, and immediate postoperative period of a pediatric population that has undergone orthotopic liver transplantation for fulminant hepatic failure in a pediatric intensive care unit. DESIGN: Retrospective review of medical records. SETTING: A pediatric intensive care unit in a children's hospital. PATIENTS: Sixty patients with fulminant hepatic failure who fulfilled King's College criteria for liver transplantation. INTERVENTION: Orthotopic liver transplantation was performed according to standard techniques. Before transplantation, patients were admitted to a pediatric intensive care unit when intensive care was required, and patients were always admitted to a pediatric intensive care unit after the operation. Measurements: A total of 20 variables were studied via univariate and multivariate analysis; statistical significance was accepted when p /=1500 g was significantly higher than for those using the Argyle catheter (p =.03). CONCLUSION: Considering the difference in gestational age for the patients weighing 1500-2500 g, we conclude that the two prongs tested are equally effective for nasal continuous positive airway pressure, but the Argyle prong is more difficult to keep in the nostrils of active patients, and nasal hyperemia, the first sign of tissue aggression, occurs more frequently among infants using this prong. PMID- 12780964 TI - Comparison of end-tidal CO2 and Paco2 in children receiving mechanical ventilation. AB - OBJECTIVE: To determine whether end-tidal CO(2) (Petco(2)) measurement provides a reliable estimate of ventilation in critically ill children who are mechanically ventilated. DESIGN: Prospective, nonrandomized, consecutive enrollment study. SETTING: A university-affiliated children's hospital pediatric intensive care unit. PATIENTS: All intubated, mechanically ventilated pediatric patients. INTERVENTIONS: All Petco(2)-Paco(2) pairs were from patients ventilated with a Servo 300 Ventilator (Siemens-Elema AB, Stockholm, Sweden). When a blood gas sample was obtained, Petco(2) as measured by a continuous mainstream Petco(2) capnograph was recorded. Measurements: The results of blood gas measurements and corresponding Petco(2) measurements were recorded. Demographic data and primary diagnosis were noted. Petco(2)-Paco(2) pairs obtained from patients with intracardiac shunts or obtained during high-frequency oscillation or extracorporeal membrane oxygenation at the time of measurement were excluded from analysis. Linear regression was used to analyze Petco(2)-Paco(2) pairs. Repeated measure analysis of variance with the mixed-model algorithm in SAS software (SAS Institute, Carey, NC) was used to analyze the trend in the Petco(2) and Paco(2) relationship. Chi-square was used to analyze categorical data. Statistical significance was considered p <.05. RESULTS: A total of 129 children were enrolled, and 1708 paired Paco(2) and Petco(2) measurements were recorded. The mean age +/- sd was 4.1 +/- 5.6 yrs. Paco(2) positively correlated with Petco(2). The linear equation for the regression analysis was y = 0.71x (95% confidence interval, 0.69-0.73) + 8.93 (95% confidence interval, 7.89-9.97), with r (2) =.716 and p <.001. The Petco(2)-Paco(2) difference was 10 mm Hg (1.33 kPa) difference between the Petco(2) and Paco(2). However, only 111 of 1068 (10%) Petco(2)-Paco(2) pairs had a difference of >10 mm Hg (1.33 kPa) in patients with a Pao(2)/Fio(2) ratio >200. Trend analysis showed the Petco(2)-Paco(2) difference increased with increasing duration of mechanical ventilation. CONCLUSION: In most intubated, mechanically ventilated infants and children, Petco(2) reliably estimates ventilation. PMID- 12780965 TI - Cisatracurium infusion for neuromuscular blockade in the pediatric intensive care unit: A dose-finding study. AB - OBJECTIVE: To evaluate the safety and efficacy of cisatracurium besylate, a neuromuscular blocking agent in infants zero to 2 yrs of age. DESIGN: An open label study to evaluate efficacy and safety of cisatracurium as a continuous infusion in infants. SETTING: A tertiary pediatric intensive care unit. PATIENTS: Eleven children, 0-2 yrs of age, requiring prolonged neuromuscular blockade. INTERVENTIONS: Cisatracurium besylate, 0.1 mg/kg, was administered as an intravenous bolus dose and repeated if necessary until a >90% neuromuscular blockade, as determined by train-of-four response, was achieved. Patients were allowed to recover to 90% blockade (I/IV twitch) after the initial bolus and were administered continuous infusion at 2 &mgr;g/kg/min. The continuous infusion rate was adjusted to maintain a train-of-four response of 0-I/IV, with an increase in the rate preceded by a bolus dose of cisatracurium besylate. An electromyographic monitor was used to measure recovery at the end of infusion, when possible. Heart rate and blood pressure were recorded after the initial bolus dose and after changes in infusion rates. Blood samples were drawn at steady-state during cisatracurium infusion at several different times during the study and at the end of infusion for measurement of plasma cisatracurium and laudanosine concentrations. MEASUREMENTS AND MAIN RESULTS: The mean infusion rate of cisatracurium besylate required to maintain train-of-four response of 0-I/iv was 5.4 +/- 3.0 &mgr;g/kg/min. The mean total duration of infusion was 64.5 +/- 36 hrs. Ten percent and complete neuromuscular recovery occurred at 26.6 +/- 10.4 and 74.8 +/- 32 mins, respectively, after discontinuation of infusion. Mean cisatracurium and laudanosine concentrations were 342.5 +/- 169 and 163.3 +/- 116 ng/mL, respectively. Four (37%) patients had undetectable (<5 ng/mL) cisatracurium concentrations at the time of 100% neuromuscular recovery (train-of four response of IV/IV or no fade at 50 mA on the electromyogram). No significant hemodynamic changes were observed during treatment with cisatracurium besylate (p <.05). CONCLUSIONS: A longer period of recovery from neuromuscular blockade was observed compared with reports of older children. Recovery from neuromuscular blockade after long-term use was not associated with any adverse events in the immediate postinfusion period. Cisatracurium besylate is a safe and effective neuromuscular blocking agent for children 0-2 yrs of age. PMID- 12780966 TI - Outcome of critically ill children before and after the establishment of a pediatric retrieval service as a component of a national strategy for pediatric intensive care. AB - OBJECTIVE: To compare outcomes of critically ill children transferred for intensive care by specialist and nonspecialist retrieval before and after the establishment of a dedicated pediatric intensive care transfer service. DESIGN: Prospective, observational study. SETTING: All hospitals that admitted children in a defined geographic region of the United Kingdom. PATIENTS: There were a total of 284 critically ill children (<16 yrs old) transferred from a district general hospital to a tertiary pediatric intensive care unit. MEASUREMENTS AND MAIN RESULTS: Age, diagnosis, Pediatric Risk of Mortality score, Therapeutic Intervention Scoring System, details of interhospital transfer, and survival to hospital discharge were analyzed for two periods, before and after the establishment of a regional transfer service. Patients were categorized as low or high risk (predicted mortality, 15%, respectively). Standardized mortality was calculated as the ratio of observed to predicted deaths in each category. After the transfer service was established, a significantly higher proportion of high-risk children were transferred by a specialist team, and there was a decreased standardized mortality rate of transferred patients (1.09 to 0.74). The greatest decrease in standardized mortality rate occurred in low-risk children transferred by nonspecialist teams (2.27 to 0.52). During the equivalent period in the same geographic region, standardized mortality rate of nontransferred children also fell in pediatric intensive care (1.59 to 0.60) and general intensive care units (1.11 to 0.81). CONCLUSIONS: We conclude that the establishment of a regional transfer service coincided with a fall in standardized mortality that reflected more general changes in intensive care performance rather than a specific benefit of a specialist transfer team. PMID- 12780967 TI - Multicenter, randomized, controlled study of porcine surfactant in severe respiratory syncytial virus-induced respiratory failure. AB - OBJECTIVE: Recently, natural exogenous surfactant replacement has been used in experimental models and clinical trials for the treatment of severe respiratory syncytial virus (RSV) disease. The present study was aimed at verifying this hypothesis and confirming the results of our previous pilot study by assessing the effect of surfactant treatment in mechanically ventilated infants with severe RSV-induced respiratory failure. DESIGN: Multicenter, randomized, controlled study. SETTING: Six pediatric intensive care units staffed by full-time intensive care physicians. PATIENTS: A total of 40 infants (20 treated and 20 controls) with RSV-induced respiratory failure requiring conventional mechanical ventilation (CMV) were randomly assigned to either exogenous surfactant (treated group) or conventional treatment (control group) over a 1-yr period. INTERVENTIONS: Fifty milligrams per kilogram of body weight of porcine-derived natural surfactant (Curosurf) was administered. The drug was instilled by means of a syringe attached to a small suction catheter inserted into the endotracheal tube down to its tip, momentarily disconnecting the patient from CMV. Main Outcome Measures: The assessment consisted of the following outcome variables: duration of CMV, length of intensive care unit stay, gas exchange, respiratory mechanics, re-treatment need, complications, and mortality. RESULTS: The two groups were similar with regard to demographics, Pediatric Risk of Mortality scores, and baseline Pao(2)/Fio(2), Paco(2), and ventilator settings. A marked increase in Pao(2)/Fio(2) and decrease in Paco(2) were observed in the treated group after surfactant administration. Hemodynamic parameters remained unchanged throughout the study period. Peak inspiratory pressure and static compliance were similar at baseline in the two groups. A decrease in peak inspiratory pressure and increase in static compliance were observed in the treated group after surfactant administration. Among surfactant-treated patients, 15 received the treatment within 24 hrs of admission, whereas the remainder (five patients) were treated later. Among children who were treated later, three needed an additional dose of surfactant. None of the children treated within 24 hrs needed an additional dose. Duration of CMV and length of stay in the intensive care unit were significantly shorter in the treated group (4.6 +/- 0.8 and 6.4 +/- 0.9 days, respectively) compared with the control group (5.8 +/- 0.7 and 8.2 +/- 1.1 days, respectively) (p <.0001). No relevant complications were observed, and all the infants survived. CONCLUSIONS: Consistent with our previous study and others, this study shows that surfactant therapy improves gas exchange and respiratory mechanics and shortens CMV and intensive care unit stay in infants with severe RSV-induced respiratory failure. PMID- 12780968 TI - Risk factors prolonging ventilation in young children after cardiac surgery: Impact of noninfectious pulmonary complications. AB - OBJECTIVE: To determine risk factors for prolonged ventilation after cardiac surgery in young children and assess the impact of noninfectious pulmonary complications on ventilatory duration. DESIGN: Retrospective case series analysis. SETTING: A tertiary pediatric cardiac center. PATIENTS: Clinical records of 222 consecutive children aged 72 hrs) to those who could be extubated at /=10 and hemodynamic instability as defined by the need for inotropes and/or >20 mL/kg fluid in a 24 hr period. INTERVENTIONS: Baseline cortisol and adrenocorticotropic hormone (ACTH) levels were measured followed by post-ACTH stimulation test cortisol levels to determine adrenal reserve. MEASUREMENTS AND MAIN RESULTS: Baseline cortisol levels ranged from 0.10 to 37.4 &mgr;g/dL (mean, 12.2 +/- 9.6). Adrenal dysfunction as defined by a baseline cortisol level of <7 &mgr;g/dL (am cortisol) or a post-ACTH stimulation level of <18 &mgr;g/dL was observed in four of the 13 patients (31%; 95% confidence interval, 5.7%,55.9%). All three of the patients with baseline cortisol levels of <7 &mgr;g/dL had low-normal ACTH levels. There was no difference in baseline PRISM scores (14.7 +/- 4.11, 12.7 +/- 2.65; p =.37) or admitting diagnoses for those with and without adrenal dysfunction. CONCLUSIONS: We conclude that some critically ill children do no mount adequate basal cortisol levels in the face of severe physiologic stress but do respond to external ACTH stimulation to better delineate the hypothalamic-pituitary-adrenal axis in critically ill children and to determine the clinical consequences of these biochemical abnormalities. PMID- 12780979 TI - Single-dose intravenous salbutamol bolus for managing children with acute severe asthma in the emergency department:Reanalysis of data. AB - OBJECTIVE: To reanalyze data from two previous studies to provide stronger evidence of benefit for early use of single-dose intravenous bolus salbutamol in children with acute severe exacerbations of asthma. Methods: Randomized, double blind, placebo-controlled trial of 84 children with acute severe asthma who presented to the emergency department of the Children's Hospital at Westmead. After clinical evaluation, patients who had severe asthma were given high-dose inhaled salbutamol and had an intravenous cannula inserted. Additional treatment consisted of intravenous methylprednisolone (1 mg/kg), oxygen (6 L/min via mask if Sao(2) was <93%). Patients were then randomized to receive an intravenous infusion of either 15 &mgr;g/kg salbutamol or saline, with clinical progress assessed hourly for 2 hrs. All patients were admitted to the hospital and clinically monitored for the proceeding 2-24 hrs, with inhaled salbutamol treatment administered in accord with the hospital's protocol. RESULTS: The intravenous salbutamol group (n = 50) demonstrated earlier clinical improvement, with earlier reduction in oxygen therapy and reduced need for ongoing inhaled salbutamol therapy by the end of phase 1 compared with the control group (n = 34). Patients in the intravenous salbutamol group were ready for discharge from the emergency department 3.7 hrs earlier than those in the control group and were ready for discharge from the hospital 9.7 hrs earlier than controls. No significant side effects were found in either group. CONCLUSION: A single-dose intravenous salbutamol bolus of 15 &mgr;g/kg administered over 10 mins in the initial treatment of children with acute severe asthma in the emergency department has the potential to shorten the duration of severe attacks and reduce overall requirements for inhaled salbutamol maintenance. PMID- 12780980 TI - Positive end-expiratory pressure and ventilation inhomogeneity in mechanically ventilated children. AB - OBJECTIVE: To obtain optimal titration of positive end-expiratory pressure (PEEP) during mechanical ventilation with functional residual capacity and ventilation homogeneity measurements. DESIGN: Experimental human and animal study. INTERVENTIONS: Functional residual capacity and five indices of uneven ventilation (alveolar mean dilution number, mean dilution number, lung clearance index, mixing ratio, and pulmonary clearance delay) were investigated in 22 children aged 0 to 14 yrs with lung disease and in seven rabbits by using a sulfur-hexafluoride wash-out technique. The children and rabbits were exposed to three different levels of PEEP (0, 5, and 10 cm H(2)O for the children and 0, 3 and 6 cm H(2)O for the rabbits). RESULTS: Functional residual capacity of the children increased from 256.9 +/- 178.6 mL (0 PEEP) to 280.0 +/- 201.1 mL (5 PEEP) and to 302.2 +/- 160.4 mL (10 PEEP, p <.001). Ventilation inhomogeneity decreased significantly in all children with increasing PEEP (p <.05). The alveolar mean dilution number decreased from 2.00 +/- 0.29 (0 PEEP) to 1.82 +/- 0.37 (5 PEEP) and to 1.66 +/- 0.34 (10 PEEP), and pulmonary clearance delay decreased from 74.9 +/- 24.2% to 66.6 +/- 38.1% and to 63.9 +/- 24.2%, respectively. The reduction in ventilation inhomogeneity was associated with an improvement in Pao(2) from 101 +/- 42 mm Hg (0 PEEP) to 122 +/- 48 mm Hg (5 PEEP) and to 126 +/- 53 mm Hg (10 PEEP). Functional residual capacity of the rabbits increased from 54.1 +/- 18.7 mL at 0 PEEP to 72.3 +/- 23.4 mL at 3 PEEP and to 93.9 +/- 27.3 mL at 10 PEEP. Alveolar mean dilution number decreased from 2.93 +/ 0.1 (0 PEEP) to 2.20 +/- 0.29 (3 PEEP) and to 1.45 +/- 0.13 (6 PEEP). CONCLUSIONS: In children with lung disease receiving ventilatory support, optimal PEEP titration can be obtained by the measurement of the functional residual capacity and ventilation distribution by using a sulfur-hexafluoride wash-in-wash out technique. PMID- 12780981 TI - National survey on the use of sedatives and neuromuscular blocking agents in the pediatric intensive care unit. AB - OBJECTIVES: To describe the sedative and neuromuscular blocking agents (NMBA) that are currently used in pediatric intensive care units across the country and to assess the use of written protocols for their use, criteria used for selecting these agents, monitoring practices, and clinicians responsible for making therapeutic decisions in the pediatric intensive care units. DESIGN: A questionnaire was mailed to pediatric attending physician members of the Society of Critical Care Medicine practicing in the United States in January 1997. A cover letter was also enclosed that explained the purpose of the survey and asked the respondent to forward the questionnaire to a colleague if unable to complete. RESULTS: A total of 176 questionnaires were returned, which represented 145 pediatric institutions across the country, for a response rate of 51%. The agents reported to be used most often for sedation were the opioids and benzodiazepines, which were used for >72 hrs. The NMBA used were vecuronium and pancuronium, which were used for >48 hrs. Newer agents such as propofol and cisatracurium were being used by some clinicians as well. Respondents primarily use the intravenous route of administration as either intermittent bolus or continuous infusion. Frequently cited indications for sedatives were anxiety, fear, and amnesia and facilitation of intubation and maintenance of mechanical ventilation for NMBA. Only 13.4% indicated using written protocols for sedatives and 26.1% for NMBA. Decisions regarding the choice of agent were usually based on clinician preference and experience and the duration of action of the agent. Respondents most often reported using clinical assessment (57%) or the Glasgow Coma Scale (47.3%) to monitor the depth or adequacy of sedation. Over 80% of those surveyed use a peripheral nerve stimulator to monitor NMBA activity. CONCLUSION: Clinicians continue to use the opioids and benzodiazepines most often for sedation in the pediatric intensive care units, but newer agents are being used more often and warrant further investigation. The use of written protocols is very low, possibly because of the lack of guidelines in the literature on pediatric intensive care unit sedation and neuromuscular blockade. Development and implementation of protocols for the selection, use, and monitoring of sedatives and NMBA through a multidisciplinary team approach may be a beneficial way to provide safe and cost effective therapy to critically ill pediatric patients. PMID- 12780982 TI - Eliminating discard volumes in neonatal and pediatric blood sampling from arterial catheters: A comparison of three simple blood-conserving aspiration techniques. AB - OBJECTIVE: To compare different blood aspiration techniques to eliminate discarding of blood in arterial blood sampling from critically ill neonates and children. DESIGN: Prospective, randomized controlled trial. SETTING: A 19-bed tertiary neonatal and pediatric intensive care unit. PATIENTS: Critically ill neonates and children with existing arterial and central venous access. INTERVENTIONS: Paired blood samples were obtained by using conventional blood discarding techniques and one of the following blood-conserving aspiration techniques: passive extracorporeal arteriovenous backflow, free passive backflow to ambient pressure, and active aspiration backflow to a distance of 10 or 20 cm proximal to the sampling port of the arterial pressure catheter. Repetitive conventional sampling served as a control and as the standard. The order of sampling was randomly allocated. We determined arterial blood gases, electrolytes, blood glucose, and hemoglobin concentration. Measurement and RESULTS: Bland-Altman bias analysis of the variability among the techniques revealed that the passive backflow and the active aspiration backflow technique with a backflow distance of 20 cm yielded identical results to repetitive conventional sampling with a standardized discard volume of 0.6 mL. In contrast, the extracorporal arteriovenous backflow technique carried the risk of overestimating blood glucose levels (mean bias, 0.96 mmol/L.). A backflow distance of 10 cm (active aspiration) proved insufficient to eliminate contamination by the catheters' flushing solution. CONCLUSIONS: At a backflow distance of 20 cm, the passive backflow and the active aspiration backflow techniques produce, with the used monitoring set, reliable and precise results in critically ill newborns and children and eliminate discard volumes. PMID- 12780983 TI - Removal of deadspace volume from arterial catheter: How muchis enough? AB - OBJECTIVE: To evaluate the amount of volume needed to be removed from arterial catheter systems to compensate for "deadspace" and to allow an accurate measurement of pH and hemoglobin (Hb). DESIGN: Twenty patients undergoing heart surgery were evaluated in a steady state after the induction of anesthesia before surgery. Six blood samples were removed from the arterial catheter, the total volume of which was 1.5 mL at 30-sec intervals and measured for pH and Hb. The first sample was then taken after removing 1.5 mL from the tubing. In subsequent samples, the volume removed before sampling increased by 0.5-mL intervals. All other samples were compared with sample number 6, in which 4 mL of volume were removed before measurements. RESULTS: The first three samples with volumes of 1.5, 2.0, and 2.5 mL before measurement were inaccurate compared with sample number 6 (p <.000), giving artificially low values for both pH and Hb. There was no significant difference between the values measured in sample numbers 4, 5, and 6 (3.0, 3.5, and 4.0 mL, respectively). CONCLUSION: The amount of volume needed to be removed before measurement from an arterial catheter system, the volume of which is 1.5 mL, is 3 mL to achieve accurate measurements of pH and Hb. Removal of less volume results in an artificially low measurement. PMID- 12780984 TI - High-frequency oscillation in children after Fontan operation. AB - OBJECTIVE: The increase in intrathoracic pressure produced by positive-pressure ventilation has been shown to have an adverse effect on pulmonary blood flow after Fontan operations. OBJECTIVE: The main objective was to compare the hemodynamic effect of high-frequency oscillation ventilation (HFOV) with intermittent positive-pressure ventilation (IPPV) in children after Fontan operations. DESIGN: Prospective study. SETTING: Department of pediatric critical care in a tertiary care, university-based children's hospital. PATIENTS: Five children (age, 2.3 +/- 0.3 yrs) in the early period after Fontan operations. INTERVENTION: After a short period of stabilization after surgery using conventional IPPV, baseline data were collected and patients were converted to HFOV. After a short period of stabilization using HFOV, a second series of measurements were performed and patients were reconnected to IPPV, after which a third series of measurements were made. Paco(2) and pH were kept unchanged throughout the study. MEASUREMENTS AND MAIN RESULTS: Hemodynamic and respiratory variables were obtained at each time point (IPPV1, HFOV, and IPPV2). The mean cardiac index at baseline (IPPV1) was 3.1 +/- 1.1 L/min/m(2); during HFOV, it was 3.2 +/- 1.2 L/min/m(2) (p =.46); and during IPPV2, it was 3.1 +/- 1.1 L/min/m(2). The mean pulmonary vascular resistance values using IPPV1, HFOV, and IPPV2 were 3.0 +/- 1.1, 2.7 +/- 1.3 (p =.21), and 2.6 +/- 1.1 Woods units/m(2), respectively. CONCLUSION: HFOV had no effect on the cardiac output or the pulmonary vascular resistance in a small group of stable patients after Fontan operations. PMID- 12780985 TI - Failed extubation after cardiac surgery in young children: Prevalence, pathogenesis, and risk factors. AB - Background: Most children who undergo congenital heart surgery require postoperative mechanical ventilation. Failed extubation (FE) may result in physiologic instability, delay, or set back of the weaning process. FE is statistically associated with prolonged mechanical ventilation. Purpose: We sought to identify frequency, pathogenesis, and risk factors for FE after congenital heart surgery in young children. SETTING: Pediatric intensive care unit. PATIENTS: Children 12 hrs after beginning BIPAP. BIPAP treatment resulted in improved ventilatory status as shown by an increase in pH from a mean of 7.26 on admission to 7.38 after 3-4 hrs and after 12 hrs, respectively. However, respiratory rate showed a steady decrease from a mean of 31.7 breaths/min on admission to 24 breaths/min at 3-4 hrs and 19 breaths/min at >12 hrs of BIPAP therapy. CO(2) tension (Pco(2)) decreased from a mean value of 54.6 mm Hg (7.28 kPa) on admission to 36.4 mm Hg (4.85 kPa) at 3-4 hrs of treatment; the mean Pco(2) after >12 hrs of treatment was 39.8 mm Hg (5.31 kPa). CONCLUSION: In three children with status asthmaticus, BIPAP seemed to improve ventilation and gas exchange, culminating in resolution of hypercarbic respiratory failure. A prospective, randomized, and controlled study is required to determine its role in pediatric status asthmaticus. PMID- 12780992 TI - Paradoxical vocal cord motion in a child presenting with cyanosis and respiratory failure. PMID- 12780993 TI - New onset of multiple organ dysfunction after receiving long-term high-dose piperacillin for outpatient treatment of osteomyelitis. AB - Toxic side effects of piperacillin occur in many organ systems and may be cumulative dose related. We report a patient receiving long-term high-dose piperacillin to accommodate convenient outpatient intravenous therapy of osteomyelitis. He developed life-threatening multiorgan dysfunction on day 20 of piperacillin administration after 49 days of antibiotics, suggesting a likely adverse drug reaction. Piperacillin should be administered according to the manufacturer's recommendations of dosage, frequency and duration, and suggested monitoring. Unorthodox treatment regimens are discouraged. PMID- 12780994 TI - Systemic inflammatory response syndrome associated with Sweet's syndrome. AB - PURPOSE: To describe the first pediatric report of systemic inflammatory response syndrome, shock, and multiple organ dysfunction syndrome associated with Sweet's syndrome. DESIGN: Case report. SETTING: Pediatric intensive care unit. PATIENTS: A patient with Sweet's syndrome and multiple organ dysfunction syndrome. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: We report the case of a 7-yr old female child with an acute nonlymphoblastic leukemia in complete remission after an autologous bone marrow transplantation, with a clinical picture of skin lesions and fever that met the criteria of Sweet's syndrome and developing systemic inflammatory response syndrome, septic shock, and multiple organ dysfunction syndrome. Her clinical condition worsened despite broad-spectrum antimicrobial therapy and standard measures of cardiovascular support. An infectious site could not be identified, and all culture results were negative. Her condition improved dramatically once steroid therapy was administered, and she made a full recovery. CONCLUSION: Although it is a rare condition, the diagnosis of Sweet's syndrome must be considered in a patient with the typical skin lesions and systemic inflammatory response syndrome. The correct diagnosis is of great clinical importance, because therapy with systemic steroids results in a fast and remarkable improvement. PMID- 12780995 TI - Acute myocardial infarction in a 12-year-old as a complication of hyperosmolar diabetic ketoacidosis. AB - OBJECTIVE: This report describes the evolution of an acute myocardial infarction in a 12-yr-old male with severe hyperosmolarity secondary to diabetic ketoacidosis. DESIGN: Case report. SETTING: Intensive care unit of a university teaching hospital. Patient: A 12-yr-old male with new-onset diabetic ketoacidosis and an abnormal electrocardiogram. MEASUREMENTS AND MAIN RESULTS: The evolution of an acute myocardial infarction in this patient is described. This includes serial electrocardiographic, echocardiographic, and enzymatic changes. Early recognition of this complication of diabetic ketoacidosis prompted immediate initiation of antithrombotic therapy and possible prevention of infarct extension. CONCLUSION: This report demonstrates the need for accurate electrocardiographic monitoring in these patients, both for arrhythmias as well as ischemic changes and the potential for myocardial injury caused by hyperosmolarity. PMID- 12780996 TI - Is the randomized, controlled trial in children an endangered species? AB - OBJECTIVE: To discuss the potential effects of current Food and Drug Administration (FDA) policy in the United States facilitating the approval of novel therapies for use in children (based on safety trials) on pediatric randomized, controlled trials in the pediatric intensive care unit setting. Method: The history of randomized, controlled trials, the FDA, and the events that led to this ruling are discussed. RESULTS: A recent FDA ruling ensures increased drug testing in children; however, it also opens the possibility for drug approval for use in children without any requirement for randomized, controlled trial testing of the efficacy of the drug on disease-specific outcomes. CONCLUSIONS: In light of the high degree of instability of pediatric intensive care unit patients, the frequent use of multiple drugs in a given patient with resultant drug-drug interactions, the risk of toxicity in critically ill infants and children with impaired drug metabolism, and age-dependent differences in physiology, clinicians must strive to be informed of the meaning of FDA approval or disapproval of the drugs they use for children. PMID- 12780997 TI - Histopathologic features of Mycobacterium ulcerans infection. AB - Because of the emergence of Buruli ulcer disease, the World Health Organization launched a Global Buruli Ulcer Initiative in 1998. This indolent skin infection is caused by Mycobacterium ulcerans. During a study of risk factors for the disease in Ghana, adequate excisional skin-biopsy specimens were obtained from 124 clinically suspicious lesions. Buruli ulcer disease was diagnosed in 78 lesions since acid-fast bacilli (AFB) were found by histopathologic examination. Lesions with other diagnoses included filariasis (3 cases), zygomycosis (2 cases), ulcerative squamous cell carcinomas (2 cases), keratin cyst (1 case), and lymph node (1 case). Thirty-seven specimens that did not show AFB were considered suspected Buruli ulcer disease cases. Necrosis of subcutaneous tissues and dermal collagen were found more frequently in AFB-positive specimens compared with specimens from suspected case-patients (p<0.001). Defining histologic criteria for a diagnosis of Buruli ulcer disease is of clinical and public health importance since it would allow earlier treatment, leading to less deforming sequelae. PMID- 12780998 TI - An ounce of prevention is a ton of work: mass antibiotic prophylaxis for anthrax, New York City, 2001. AB - Protocols for mass antibiotic prophylaxis against anthrax were under development in New York City beginning in early 1999. This groundwork allowed the city's Department of Health to rapidly respond in 2001 to six situations in which cases were identified or anthrax spores were found. The key aspects of planning and lessons learned from each of these mass prophylaxis operations are reviewed. Antibiotic distribution was facilitated by limiting medical histories to issues relevant to prescribing prophylactic antibiotic therapy, formatting medical records to facilitate rapid decision making, and separating each component activity into discrete work stations. Successful implementation of mass prophylaxis operations was characterized by clarity of mission and eligibility criteria, well-defined lines of authority and responsibilities, effective communication, collaboration among city agencies (including law enforcement), and coordination of staffing and supplies. This model can be adapted for future planning needs including possible attacks with other bioterrorism agents, such as smallpox. PMID- 12780999 TI - Inactivation of Bacillus anthracis spores. AB - After the intentional release of Bacillus anthracis through the U.S. Postal Service in the fall of 2001, many environments were contaminated with B. anthracis spores, and frequent inquiries were made regarding the science of destroying these spores. We conducted a survey of the literature that had potential application to the inactivation of B. anthracis spores. This article provides a tabular summary of the results. PMID- 12781000 TI - Children with respiratory disease associated with metapneumovirus in Hong Kong. AB - Human metapneumovirus (HMPV) is a newly discovered pathogen thought to be associated with respiratory disease. We report the results of a study of 587 children hospitalized with respiratory infection over a 13-month period. HMPV was detected in the nasopharyngeal aspirates from 32 (5.5%) children by reverse transcription-polymerase chain reaction. HMPV infection was associated with clinical diagnoses of pneumonia (36%), asthma exacerbation (23%), or acute bronchiolitis (10%). When compared to those with respiratory syncytial virus infection, children with HMPV infection were older, and wheezing was more likely to represent asthma exacerbation rather than acute bronchiolitis. HMPV viral activity peaked during the spring-summer period in Hong Kong. Phylogenetically, all HMPV virus strains from Hong Kong belonged to one of the two genetic lineages previously described. HMPV contributed to 441.6 hospital admissions per 100,000 population <6 years of age. PMID- 12781001 TI - Human metapneumovirus infections in hospitalized children. AB - We evaluated the percentage of hospitalizations for acute respiratory tract infections in children < or =3 years of age attributable to human metapneumovirus (HMPV) and other respiratory viruses in a prospective study during winter and spring 2002. We used real-time polymerase chain assays and other conventional diagnostic methods to detect HMPV, human respiratory syncytial virus (HRSV), and influenza viruses in nasopharyngeal aspirates of children. HMPV was detected in 12 (6%) of the 208 children hospitalized for acute respiratory tract infections, HRSV in 118 (57%), and influenza A in 49 (24%). Bronchiolitis was diagnosed in 8 (68%) and pneumonitis in 2 (17%) of HMPV-infected children; of those with HRSV infection, bronchiolitiss was diagnosed in 99 (84%) and pneumonitis in 30 (25%). None of the HMPV-infected children was admitted to an intensive-care unit, whereas 15% of those with HRSV or influenza A infections were admitted. HMPV is an important cause of illness in young children with a similar, although less severe, clinical presentation to that of HRSV. PMID- 12781002 TI - Dead bird clusters as an early warning system for West Nile virus activity. AB - An early warning system for West Nile virus (WNV) outbreaks could provide a basis for targeted public education and surveillance activities as well as more timely larval and adult mosquito control. We adapted the spatial scan statistic for prospective detection of infectious disease outbreaks, applied the results to data on dead birds reported from New York City in 2000, and reviewed its utility in providing an early warning of WNV activity in 2001. Prospective geographic cluster analysis of dead bird reports may provide early warning of increasing viral activity in birds and mosquitoes, allowing jurisdictions to triage limited mosquito-collection and laboratory resources and more effectively prevent human disease caused by the virus. This adaptation of the scan statistic could also be useful in other infectious disease surveillance systems, including those for bioterrorism. PMID- 12781003 TI - Gnathostomiasis: an emerging imported disease. AB - As the scope of international travel expands, an increasing number of travelers are coming into contact with helminthic parasites rarely seen outside the tropics. As a result, the occurrence of Gnathostoma spinigerum infection leading to the clinical syndrome gnathostomiasis is increasing. In areas where Gnathostoma is not endemic, few clinicians are familiar with this disease. To highlight this underdiagnosed parasitic infection, we describe a case series of patients with gnathostomiasis who were treated during a 12-month period at the Hospital for Tropical Diseases, London. PMID- 12781004 TI - Clinical implications of varying degrees of vancomycin susceptibility in methicillin-resistant Staphylococcus aureus bacteremia. AB - We conducted a retrospective study of the clinical aspects of bacteremia caused by methicillin-resistant Staphylococcus aureus (MRSA) with heterogeneously reduced susceptibility to vancomycin. Bloodstream MRSA isolates were screened for reduced susceptibility by using brain-heart infusion agar, including 4 mg/L vancomycin with and without 4% NaCl. Patients whose isolates exhibited growth (case-patients) were compared with those whose isolates did not (controls) for demographics, coexisting chronic conditions, hospital events, antibiotic exposures, and outcomes. Sixty-one (41%) of 149 isolates exhibited growth. Subclones from 46 (75%) of these had a higher MIC of vancomycin than did their parent isolates. No isolates met criteria for vancomycin heteroresistance. No differences in potential predictors or in outcomes were found between case patients and controls. These data show that patients with vancomycin-susceptible MRSA bacteremia have similar baseline clinical features and outcomes whether or not their bacterial isolates exhibit growth on screening media containing vancomycin. PMID- 12781005 TI - Serogroup W-135 meningococcal disease during the Hajj, 2000. AB - An outbreak of serogroup W-135 meningococcal disease occurred during the 2000 Hajj in Saudi Arabia. Disease was reported worldwide in Hajj pilgrims and their close contacts; however, most cases were identified in Saudi Arabia. Trends in Saudi meningococcal disease were evaluated and the epidemiology of Saudi cases from this outbreak described. Saudi national meningococcal disease incidence data for 1990 to 2000 were reviewed; cases from January 24 to June 5, 2000, were retrospectively reviewed. The 2000 Hajj outbreak consisted of distinct serogroup A and serogroup W-135 outbreaks. Of 253 identified cases in Saudi Arabia, 161 (64%) had serogroup identification; serogroups W-135 and A caused 93 (37%) and 60 (24%) cases with attack rates of 9 and 6 cases per 100,000 population, respectively. The 2000 Hajj outbreak was the first large serogroup W-135 meningococcal disease outbreak identified worldwide. Enhanced surveillance for serogroup W-135, especially in Africa, is essential to control this emerging epidemic disease. PMID- 12781006 TI - Molecular subtyping to detect human listeriosis clusters. AB - We analyzed the diversity (Simpson's Index, D) and distribution of Listeria monocytogenes in human listeriosis cases in New York State (excluding New York City) from November 1996 to June 2000 by using automated ribotyping and pulsed field gel electrophoresis (PFGE). We applied a scan statistic (p IHPH-OLT A > IHPH OLT B rats. The level of plasma PGI(2) in IHPH rats was significantly elevated compared with NL rats, while PGI(2) in IHPH-OLT A and B rats was found to be lower than in IHPH rats (P < 0.05). There was no obvious difference in PGI(2) between IHPH-OLT B and NL rats. Vasoconstrictors including ET and TXA(2) were found elevated in IHPH-OLT rats. CONCLUSIONS: OLT does not induce postoperative hyperhemodynamics per se. Vasodilators including NO and Glu, especially NO, play an important role in the hyperhemodynamics of IHPH and IHPH-OLT rats. The results of the present study demonstrate that the persistence of systemic and splanchnic hyperkinetic circulation in the early stages after OLT may result from those non eliminated factors that caused hyperhemodynamics in liver cirrhosis patients with portal hypertension before OLT. PMID- 12781048 TI - Neurotization of oculomotor, trochlear and abducent nerves in skull base surgery. AB - OBJECTIVE: To anatomically reconstruct the oculomotor nerve, trochlear nerve, and abducent nerve by skull base surgery. METHODS: Seventeen cranial nerves (three oculomotor nerves, eight trochlear nerves and six abducent nerves) were injured and anatomically reconstructed in thirteen skull base operations during a period from 1994 to 2000. Repair techniques included end-to-end neurosuture or fibrin glue adhesion, graft neurosuture or fibrin glue adhesion. The relationships between repair techniques and functional recovery and the related factors were analyzed. RESULTS: Functional recovery began from 3 to 8 months after surgery. During a follow-up period of 4 months to 6 years, complete recovery of function was observed in 6 trochlear nerves (75%) and 4 abducent nerves (67%), while partial functional recovery was observed in the other cranial nerves including 2 trochlear nerves, 2 abducent nerves, and 3 oculomotor nerves. CONCLUSIONS: Complete or partial functional recovery could be expected after anatomical neurotization of an injured oculomotor, trochlear or abducent nerve. Our study demonstrated that, in terms of functional recovery, trochlear and abducent nerves are more responsive than oculomotor nerves, and that end-to-end reconstruction is more efficient than graft reconstruction. These results encourage us to perform reconstruction for a separated cranial nerve as often as possible during skull base surgery. PMID- 12781049 TI - Combined therapy of methylprednisolone and brain-derived neurotrophic factor promotes axonal regeneration and functional recovery after spinal cord injury in rats. AB - OBJECTIVE: To investigate the effects of combination therapy with methylprednisolone (MP) and brain-derived neurotrophic factor (BDNF) on axonal remyelination and functional recovery after spinal cord injury in rats. METHODS: Forty-five rats were randomly divided into three groups: Group A received MP and BDNF; group B received MP and cerebrospinal fluid (CSF); and group C received CSF only. Contusion injury to adult rat spinal cord was produced at the T(10) vertebra level followed by immediate intravenous MP or CSF, and was thereafter infused intrathecally with BDNF or CSF for 6 weeks. Axonal remyelination and functional recovery was observed using RT-PCR, immunohistochemistry and open field locomotion. RESULTS: An increase of 28.4% +/- 2.3% in the expression of proteolipid protein (PLP) gene, an endogenous indicator of axonal remyelination, was demonstrated in group A 24 hours after injury. Ten weeks later, there were significant decreases in hematogenous inflammatory cellular infiltration in groups A and B compared to C (P < 0.05). Concomitantly, a significant amount of axonal remyelination was observed in group A compared to groups B and C (P < 0.05). Furthermore, combination therapy using MP and BDNF in group A resulted in stimulation of hindlimb activity as well as improvement in the rate of functional recovery in open field locomotion (P < 0.05). CONCLUSIONS: Combined therapy of MP and BDNF can improve functional recovery through mechanisms that include attenuating inflammatory cellular infiltration and enhancing axonal remyelination at the injury site. Such a combination may be an effective approach for treatment of spinal cord injury. PMID- 12781050 TI - Preparation of collagen-based materials for wound dressing. AB - OBJECTIVE: To describe the methods which were used to develop collagen-based materials for wound dressing. METHODS: Fresh frozen bovine tendon was treated with 0.05 mol/L acetic acid at pH 3.2 for 48-72 hours, homogenized, filtered, mixed with 8% chondroitin sulphate, for creating a deaerated 1.5%-2.5% collagen solution. The solution was lyophilized in either a pre-frozen or non-pre-frozen mould. The collagen sponge was then cross-linked with 0.25% glutaraldehyde for 24 hours. Three other types of wound dressings were developed using a similar method: collagen membrane with a polyurethane membrane onlay, polyurethane-coated collagen membrane and collagen membrane on gauze. RESULTS: It was demonstrated that the use of frozen bovine tendon was stable, and that the prepared collagen sponge contained pores of 50-400 microm in diameter. CONCLUSIONS: Collagen could be used as wound dressing. PMID- 12781051 TI - Pathological observation of brain arteries and spontaneous aneurysms in hypertensive rats. AB - OBJECTIVE: To investigate the role of hypertension in the pathogenesis of cerebral aneurysms in rats. METHODS: Twenty spontaneous hypertensive rats (SHR) and 10 Wistar-Kyoto rats (WKY) were included in this observational study. Animals were fed with normal diet and drinking water. No experimental modifications were undertaken in either group. They were sacrificed at one year of age, the bifurcations of the circle of Willis were dissected and longitudinal serial sections were prepared for light microscopic and transmission electron microscopic study. RESULTS: In the SHR group, 2 of the 20 rats formed an aneurysm respectively at the bifurcations of the basilar artery. As revealed by electron microscopy, injury at the bifurcation of the artery first occurred on the steeper side of the intimal pad. Furthermore, loss of endothelial cells, small depressions on the intima, disruptive internal elastic lamina and lymphocytes or red blood cells infiltration were noted at the steeper side of the intimal pad. No significant changes were observed in WKY group. CONCLUSIONS: Cerebral aneurysms can form spontaneously in SHR without ligation of the common carotid artery and without a diet containing beta-aminoproprionitrile. Long-standing systemic arterial hypertension is one of the etiological factors that contributes to aneurysm formation in SHR rats. PMID- 12781052 TI - Differentiation of rat embryonic neural stem cells promoted by co-cultured Schwann cells. AB - OBJECTIVE: To explore the factors which induce differentiation of embryonic neural stem cells. METHODS: Rat embryonic neural stem cells were co-cultured with newborn rat Schwann cells in serum-free medium. The phenotype and specific markers including tubulin-beta, glial fibrillary acidic protein (GFAP) and galactorcerebroside (GalC), were demonstrated by phase contrast microscopy and double immunofluorescence staining. RESULTS: Overall, 80% +/- 5% of neural stem cells protruded several elongated processes and expressed tubulin-beta antigen at high levels, while 20 +/- 3% of them protruded several short processes and were GalC or GFAP positive. CONCLUSION: The factors secreted by Schwann cells could induce rat embryonic neural stem cell to differentiate. PMID- 12781053 TI - Anti-CD25 monoclonal antibody modulates cytokine expression and prolongs allografts survival in rats cardiac transplantation. AB - OBJECTIVE: To investigate the role of anti- interleukin-2 receptor (CD25) monoclonal antibody in the regulation of cytokine mRNA expression of IL-1beta, IL 2, CD25, IL-4, IL-5, IL-6, IL-10, tumour necrosis factor-alpha (TNFalpha), and interferon-gamma (IFNgamma) in cardiac allografts to elucidate its immunological mechanism and role in rats that have undergone cardiac transplantation. METHODS: These in vivo studies were conducted using a rat MHC mismatch SD to Wistar heterotopic cardiac transplant model. Simulect, an anti-CD25 antibody, was used to prevent allograft rejection. An increase in the rate of allograft survival was observed. Rats were sacrificed on day 1, 3, 5, 7, 9, 11, 14 post-transplantation and hearts were harvested for further study. Cytokine mRNA expression was determined by semiquantitative RT-PCR. RESULTS: In the control group, cardiac allografts were rejected at 8.3 +/- 1.7 days after transplantation (x +/- s). The rats who received CsA rejected the cardiac allograft at 26.4 +/- 5.7 days post transplant. Allograft survival of Simulect-treated rats was 29.2 +/- 7.1 days (P < 0.05 vs controls). Rats treated with simulect and CsA had the longest survival of 55.0 +/- 11.6 days (P < 0.001 vs controls). CD25 mRNA expression in the heart tissue samples of treated rats was undetectable or very weak. However, the untreated group, CD25 expression increased, although anti-CD25 decreased this CD25 expression in the heart graft. Furthermore, in untreated allografts, IL-2, TNFalpha and IFN-gamma were strongly expressed, an effect that markedly decreased after simulect treatment. Finally, IL-4, IL-5, IL-6 and IL-10 expression was strong in anti-CD25-treated allografts. CONCLUSIONS: These results suggest that anti-CD25 antibody treatment may not only neutralize CD25 activity but also play a role in altering cytokine mRNA expression and prolong the survival of allografts. PMID- 12781054 TI - Graft patency in off-pump and conventional coronary artery bypass grafting for treatment of triple vessel coronary disease. AB - OBJECTIVE: To compare graft patency in off-pump and conventional coronary artery bypass grafting by using the transit time flow meter in the treatment of triple vessel coronary artery disease. METHODS: Between June 2000 and April 2001, 60 patients with triple vessel coronary artery disease underwent coronary artery bypass grafting. They were divided into two groups: off-pump and conventional coronary artery bypass. All completed grafts were tested intraoperatively using Transit Time Flow Measurement (TTFM). Preoperative and postoperative variables of the two groups were also compared. RESULTS: There were no significant differences in sex, age, weight, acute or remote myocardial infarction, hypertension, diabetes and type of bypass grafts between the two groups. The number of bypass grafts and the assisted respiratory time of the off-pump coronary artery bypass grafting (OPCAB) group were significantly less than those of the conventional coronary artery bypass grafting (CCABG) group. The flow and pulsatile index (PI) of the left anterior descending artery bypass grafts and the right coronary artery bypass grafts were not significantly different between the OPCAB and CCABG groups. The flow of OM in the CCABG group with the multiple anastomosis site of sequential grafts was higher than that in the OPCAB group. Diffused narrow coronary artery bypass grafts in both groups had less flow. CONCLUSION: No significant differences in graft patency were observed in patients with triple vessel coronary artery disease who had undergone OPCAB or CCABG. PMID- 12781055 TI - Prophylactic effect of Ca2+ -deficient artificial perilymph perfusion on noise induced hearing loss. AB - OBJECTIVE: To investigate the prophylactic effect of low calcium concentration perilymph on noise-induced hearing loss. METHODS: Forty guinea pigs with normal hearing weighing 250-350 g were assigned to five groups (8 in each group): (1) Ca(2+)-deficient perilymph perfusion (CDP) for 2 h; (2) white noise (120 dB SPL) exposure (WNE) only for 1 h, (3) combination of calcium-deficient perilymph perfusion and white noise (120 dB SPL) exposure (WNE + CDP); (4) normal artificial perilymph (NAP) perfusion for 2 h; and (5) white noise exposure + normal artificial perilymph perfusion (WNE + NAP) for 2 h. Compound action potentials (CAP) evoked by click was recorded from round window every 15 min. The cochleae from 5 animals in each group were examined with scanning electron microscope. RESULTS: The CAP for group 1 experienced a threshold shift (TS) of 15 26 dB, while group 2 yielded a 46-59 dB TS and group 3 a 37-45 dB TS; no threshold shift occurred in group 4. The CAP TS in group 5 was 33-64 dB. The CAP TS of group 3 was less than that of group 2. After one hour of noise exposure, the CAP TS of group 3 were 45.92 +/- 2.90 dB and 59.30 +/- 3.95 dB in group 2. There were significant differences (P < 0.05) between groups 3 and 2. The CAP TS of group 3 was less than that of group 5 at the points of 1, 1.5 and 2 h after noise exposure. There was a significant difference between groups 3 and 5 (P < 0.01). Stereocilia of 89 OHC(3) were in disarray in five cochleae after noise exposure in group 2. The cuticular plates of 8 OHC(2),3 sank and the stereocilia became fused in only one animal cochlea after noise exposure in group 3 combined with low calcium perilymph perfusion. CONCLUSIONS: Low calcium concentration appears to participate in preventing noise-induced hearing loss and the rising of calcium concentrations in inner hair cells after noise exposure, which may have been due to the opening of calcium channels in inner hair cells during noise exposure. The mechanism of the prophylactic effect might be caused by a lower calcium concentration in inner hair cells in the cochlea attenuating the influence of noise exposure on hearing loss; calcium deficient perilymph perfusion prevented calcium accumulation in inner hair cells of the cochlea. The motility of the OHCs might be partially inhibited by low calcium concentration that reduced noise-induced hearing loss in turn. PMID- 12781056 TI - Optical coherence tomography's diagnostic value in evaluating surgical impact on idiopathic macular hole. AB - OBJECTIVE: To identify the characteristics of and evaluate surgical impact on idiopathic macular hole (IMH) by using an optical coherence tomography (OCT) scanner. METHODS: Sixty-five cases (70 eyes) experiencing IMH were examined using OCT, then graded by their clinical characteristics. Nineteen cases (19 eyes) were scanned and measured using OCT before and after surgery. RESULTS: Of the 70 eyes,the number of stage I-IV macular holes were 11, 12, 36 and 11, respectively. For stage I holes, the OCT images revealed flattened or nonexistent fovea and minimally reflective space within or beneath the neurosensory retina; stage II holes appeared to be full-sized with attached operculum and surrounding edema; stage III holes were also full-sized with surrounding edema; finally, stage IV holes were full-sized and completely separated the posterior hyaloid membrane from the retina. Through quantitative measurements, OCT determined that the values for mean hole diameter, mean halo diameter and mean thickness of the hole's edge were reduced from 570.95 +/- 265.59 to 337.05 +/- 335.95 microm, 1043.53 +/- 278.8 to 695.00 +/- 483.00 microm and 389.78 +/- 60.58 to 298.78 +/- 109.80 microm, respectively in 19 IMH cases after surgery. In 17 eyes, the holes or halos eventually closed or were reduced in size, or the edges of the holes thinned out. The anatomic successful rate of the surgery was 89%. CONCLUSION: OCT can exhibit the characteristics of IMH and measure the diameter of holes quantitatively. This method can also judge the surgical impacts of IMH objectively, accurately and effectively. PMID- 12781057 TI - Identification of genes with consistent expression alteration pattern in ACC-2 and ACC-M cells by cDNA array. AB - OBJECTIVE: To investigate the molecular events and metastasis-related genes in ACC-2 and ACC-M cells of adenoid cystic carcinoma. METHODS: Adenoid cystic carcinoma cell line ACC-2 and a sample of adenoic cystic carcinoma cell clones highly metastatic to the lung (ACC-M) were investigated. ACC-2 and ACC-M cells were cultured and collected. Total RNA was extracted using standard Trizol RNA isolation protocol. The poly A mRNA was purified and labeled in reverse transcription using M-MLV reverse transcriptase in the presence of Easytides deoxyadenosin 5' triphosphate [alpha-(33)p]. A cDNA array was assembled with 7675 EST clones which represented the same number of independent single genes. Prepared nylon membranes were hybridized with the [alpha (33)p]-dATP labeled mRNA from ACC-2 and ACC-M cells. Membranes were exposed to phosphor screen. RESULTS: The high-through put analysis of gene expression pattern was obtained from ACC-2 and ACC-M cells by the hybridization of the cDNA array. The difference of parallel gene expression was analyzed. Genes were clustered according to their expression level in the ACC-M compared with ACC-2 cells. According to each gene's ratio of expression level, there were 17 genes which were upregulated with ratios over 3.0, and there were 12 genes which were downregulated with ratios below 0.33 (1/3.0 = 0.33). CONCLUSIONS: Significantly different expression patterns between ACC-2 and ACC-M by cDNA array were observed. The differences lie in signal pathways, tumor antigens, immune molecular and some unknown genes. PMID- 12781058 TI - Omega-3 fatty acids and non-communicable diseases. AB - OBJECTIVE: To review the relation between dietary omega-3 polyunsaturated fatty acids (omega-3 PUFA) and non-communicable diseases. METHOD: Data were collected from scientific journals and conference publications, MEDLINE (1979 - 2002) and current content which included 68 prospective, cross-sectional, case control and dietary-intervention studies. Scientific paper selections were based on the association between omega-3 PUFA and non-communicable diseases. RESULTS: omega-3 PUFA has beneficial effects on increasing heart rate variability, decreasing the risk of stroke, reducing both systolic and diastolic blood pressure, insulin resistance and glucose metabolism. Long chain omega-3 PUFA has anti-cancer and anti-inflammatory activities. omega-3 PUFA has also been reported to have a beneficial effect on attention-deficit/hyperactivity disorder and schizophrenia, and may be effective in managing depression in adults. CONCLUSIONS: Results from epidemiological and dietary intervention studies have shown that omega-3 PUFA represent powerfully a class of bioactive compounds and that dietary intake of omega-3 PUFA plays a critical role in human health in relation to non communicable diseases. PMID- 12781059 TI - Effects of leukotriene receptor antagonist on chronic obstructive [correction of obstractive] pulmonary disease induced pulmonary hypertension. AB - OBJECTIVES: To assess the hemodynamic, oxygen-dynamic and ventilative effects of Zafirlukast in chronic obstructive pulmonary disease (COPD) induced chronic cor pulmonale at acute exacerbation stage and the mechanisms of Zafirlukast efficacy. METHODS: Eleven cases of chronic cor pulmonale at acute exacerbation were examined using Swan-Ganz catheter and peripheral intra-artery catheter. The hemodynamic, oxygen-dynamic parameters and respiratory rate, plasma endothelium-1 (ET-1) level, and urea leukotriene E(4) (LTE(4)) level were measured before and at the 1st, 3rd, 5th, 7th, 9th, 12th hour after taking 40 mg Zafirlukast orally. Arterial and mixed venous blood gas analyses were done correspondingly. RESULTS: The average pulmonary arterial pressure (mPAP) and pulmonary vascular resistance index (PVRI) were lowered at the 3rd hour after taking Zafirlukast by 23% and 36.5%, respectively. They returned to the baseline around 12th hour. Respiratory rate decreased significantly within the 3rd-7th hour after taking Zafirlukast. LTE(4) and ET-1 levels lowered at the 3rd hour and showed a positive correlation with change of mPAP. CONCLUSIONS: Zafirlukast can reduce mPAP, pulmonary vascular resistance (PVR) and does not affect the ambulatory blood pressure monitoring (ABPM) and oxygenation in cases of chronic cor pulmonale at acute exacerbation stage. Zafirlukast may play a role as an alternative to decrease PAP in COPD patients. PMID- 12781060 TI - The role of CTLA4-Ig in a mouse model against allergic asthma. AB - OBJECTIVE: To investigate CTLA4-Ig's potential role in therapy for allergic asthma by blocking B7/CD28 interactions with cytotoxic T lymphocyte antigen 4-Ig (CTLA4-Ig). METHODS: We divided BALB/C mice into the three groups: Sham/Sham (control), ovalbumin (OVA)/OVA and mCTLA4-Ig. Blood, bronchoalveolar lavage, histology and determination of cytokines were performed 24 hours after airway challenge. RESULTS: In the OVA/OVA group, the number of cells, the percentage of inflamed cells and the level of IL-4 in BALF were increased. Airways in our murine model for allergic asthma underwent pathological changes, which were significantly reduced after treatment with mCTLA4-Ig. CONCLUSION: Blockage of co stimulation with mCTLA4-Ig can inhibit allergy-specific response of T cells, and asthmatic response as well. PMID- 12781061 TI - Primary culture and identification of sinoatrial node cells from newborn rat. AB - OBJECTIVE: To establish a reliable approach to primary culture and identification of sinoatrial node (SAN) cells. METHODS: The SAN cells were cultured from SAN tissue removed from neonatal Wistar rats and purified with differential attachment and 5'-bromodeoxyuridine (BrdU) treatment. The obtained cells were morphologically observed with inverted microscopy and transmission electron microscopy. Its action potential was recorded using electrophysiological methods. RESULTS: Three distinctly different cells were observed in the cultured SAN cells: spindle, triangle and irregular. Of these, the spindle cells comprised the greatest proportion, with their shape, structure and electrophysiological characteristics consistent with those of the pacemaker cells of SAN. The triangle cells were similar in features to the similarly shaped myocytes located in the atrial myocardium. CONCLUSIONS: The culture method of differential attachment combined with BrdU treatment is a reliable approach to growing SAN cells. Of the cells cultured from SAN, the spindle cells appear to function as pacemaker cells. PMID- 12781062 TI - Clinical significance of serum cardiac troponin T in patients with congestive heart failure. AB - OBJECTIVE: To determine whether the level of serum cardiac troponin T (cTnT) was increased in patients with congestive heart failure (CHF). METHODS: This study consisted of 265 patients with CHF and 75 healthy people. Serum cTnT was measured by electrochemiluminescence immunoassay using an Elecsys 1010 automatic analyzer. RESULTS: cTnT concentration was 0.181 +/- 0.536 ng/mL in CHF patients and 0.003 +/- 0.001 ng/mL in controls (P < 0.001). Patients were categorized according to the levels of heart function and left ventricular ejection fraction (LVEF). In the first group consisting of 105 patients with LVEF 35%, cTnT was 0.07 +/- 0.0 5 ng/mL (P < 0.01). In patients with NYHA class I, II, III and IV, cTnT values were 0.062 +/- 0.022 ng/mL, 0.113 +/- 0.121 mg/mL, 0.191 +/- 0.231 mg/ml and 0.384 +/- 0.211 mg/mL, respectively (class I vs class II P > 0.05, class II vs class III P < 0.01, class III vs class IV P < 0.01). A negative correlation was observed between serum cTnT concentration and LVEF in 265 patients with CHF (r = -0.493, P < 0.001). CONCLUSIONS: This study shows that the level of serum cTnT is increased in patients with CHF and that the increased level indicates the severity of CHF. PMID- 12781063 TI - Effects of vitamin E on the proliferation and collagen synthesis of rat hepatic stellate cells treated with IL-2 or TNF-alpha. AB - OBJECTIVE: To study the effects of vitamin E on the proliferation and collagen synthesis of rat hepatic stellate cells treated with interleukin-2 (IL-2) or tumor necrosis factor-alpha (TNF-alpha). METHODS: Hepatic stellate cells were isolated from male Sprague-Dawley rats by using modified Friedman's method. Using the isolated cells cultured and treated with IL-2 or TNF-alpha, we studied the effects of vitamin E on their proliferation and collagen synthesis through an (3)H-thymidine and (3)H-proline incorporation assay, as well as through observation of these cells under a contrary phase microscope. RESULTS: Adding IL 2 increased the both proliferation and collagen synthesis of hepatic stellate cells. Their proliferation was also increased by the addition of TNF-alpha, although it decreased collagen synthesis. Vitamin E had marked inhibitory effects on the ability of cells treated with IL-2 or TNF-alpha to reproduce or synthesize collagen. CONCLUSION: Vitamin E can inhibit the proliferation and collagen synthesis of hepatic stellate cells. It is possible that vitamin E affects liver fibrosis through these activities. PMID- 12781065 TI - The potential role of IGF-I receptor mRNA in rats with diabetic retinopathy. AB - OBJECTIVE: To evaluate the potential role of insulin-like growth factor-1 receptor mRNA (IGF-IR mRNA) in the onset and development of retinopathy in diabetic rats. METHODS: A diabetic model was duplicated in Wistar rats. The early changes in the retina were examined using light and transmission electron microscopy. Expression of IGF-IR mRNA was analyzed using in situ hybridization. RESULTS: Weak expression of IGF-IR mRNA (5%) was found in retinas of normal rats, but was significantly increased (15% and 18%) in the retinas of diabetic rats after 3 and 6 months of diabetes (P < 0.01). In situ hybridization and morphological study demonstrated that there was a positive correlation between IGF-IR mRNA expression and retinal changes at various stages. CONCLUSION: Increased IGF-IR mRNA might play an important role in the onset and development of diabetic retinopathy. PMID- 12781064 TI - Coagulation and fibrinolytic activity in patients with acute cerebral infarction. AB - OBJECTIVE: To measure the concentration of D-dimer (DD), tissue plasminogen activator (t-PA), plasminogen activator inhibitor-1 (PAI-1) and plasminogen (PLG) activity in plasma and cerebrospinal fluid in patients with acute cerebral infarction and to investigate their clinical significance. METHODS: The concentrations of D-dimer, t-PA, and PAI-1 in plasma and cerebrospinal fluid in patients were measured by enzyme-linked immunosorbent assay (ELISA). The PLG biological activity was detected using the chromophore method. The results were compared with those of the controls. RESULTS: The concentrations of D-dimer, t-PA and PAI-1 in cerebrospinal fluid and plasma in patients with acute cerebral infarction were much higher than those of normal subjects (P < 0.01). Conversely, the level of PLG activity was significantly lower in the patients than in the controls (P < 0.01). CONCLUSION: Hypercoagulability and secondary hyperfibrinolysis exist in patients with acute cerebral infarction. PMID- 12781066 TI - Limitations to the use of the CA-125 antigen level in ovarian cancer. PMID- 12781067 TI - Early results from randomized phase III trial of imatinib mesylate for gastrointestinal stromal tumors. PMID- 12781068 TI - Recent advances in radiotherapy for soft-tissue sarcoma. AB - The use of radiotherapy in soft-tissue sarcoma continues to evolve. This review focuses on how current management is influenced by the most recent publications in the field. In particular, experience of patients treated without radiotherapy permits guidelines to be established that define a subset of patients who do not need radiotherapy to achieve sarcoma cure with good function. Strategies for radiotherapy delivery are discussed, including the most recent prospective results from a trial of preoperative and postoperative radiotherapy, with particular emphasis on randomized data. Also, the definitions for adequacy of surgical excision and the ability to achieve high rates of local control when margins are minimally positive are integrated into a planned approach. The difficult problem of retroperitoneal sarcoma is discussed, as are chemoradiotherapy protocols that may enhance local and systemic outcome. Finally, the potential for image-guided radiotherapy, enhanced targeting, and better radiotherapy delivery in the contemporary era is addressed. PMID- 12781070 TI - Recent advances in the management of gastrointestinal stromal tumors. AB - Gastrointestinal stromal tumors (GISTs), the most common mesenchymal tumors of the intestinal tract, are characterized by the expression of KIT, also known as CD117. Increasingly, primary tumors from novel sites are being described. The KIT gene is commonly mutated, causing constitutive activation of the protein and aberrant growth. Recently, tumors with platelet-derived growth factor receptor (PDGFR)-a mutations have been described in GISTs with wild-type KIT. Factors that predict for an unfavorable outcome are being recognized. A specific molecularly targeted drug, imatinib mesylate, has altered the treatment of this disease. The results of phase I, II, and III clinical trials have consistently demonstrated activity of this agent and elucidated the patient and tumor characteristics associated with response to imatinib. The current challenge in caring for these patients is to identify the appropriate clinical setting for treatment with imatinib and to define the approach to patients whose tumors are insensitive or refractory to imatinib. PMID- 12781069 TI - Immunity against soft-tissue sarcomas. AB - Recent advances in basic medical sciences have led to a deeper understanding of the molecular characteristics of soft-tissue sarcomas. Likewise, novel technologies have led to a better appreciation of the relationship between an antigenic stimulus and the subsequent immune response against the antigen. In the past few years, the intersection of the understanding of the immune system and the knowledge of sarcoma biology has become apparent. As seen with other forms of cancer, there is a detectable autologous immune response against sarcomas. It is the hope of many investigators that the hints of a tumor-specific immune response will be enough to generate a signal that can be amplified and directed against the host sarcoma. The data regarding the initial evidence of immune responses against sarcomas are reviewed in the context of current or potential clinical studies. PMID- 12781072 TI - Induction chemotherapy in early-stage non-small-cell lung cancer. PMID- 12781071 TI - Molecular targets in therapy for human soft-tissue and bone sarcomas. AB - Sarcomas represent a heterogeneous group of tumors with different natural histories and therapeutic approaches. Recent discoveries have identified molecular alterations in the pathogenesis of these tumors that lead to distinct effects on sarcoma cell biology. These tumor cell characteristics include independence from growth factors, evasion of apoptosis, and maintenance of genomic integrity. Inhibition of these molecular alterations represents a therapeutic opportunity to reverse the biologic basis of tumor formation in soft tissue sarcomas and bone tumors. PMID- 12781073 TI - Lung cancer screening. AB - Low-dose CT screening for lung cancer is a complex and controversial topic. This article reviews the history of lung cancer screening trials and addresses the principles and confounding biases associated with screening. Chest radiography was initially used for lung cancer screening in the 1970s. In the mid-1990s helical single-detector CT came into use, followed by helical multidetector CT, the current method of screening. Results from prevalence studies and a few single arm incidence studies have raised concerns about overdiagnosis and the high rate of nodule detection. Follow-up studies and further investigation are needed. To this end, a randomized, controlled trial sponsored by the National Cancer Institute is underway to evaluate disease-specific mortality. PMID- 12781074 TI - Concurrent chemoradiotherapy for inoperable stage III non-small-cell lung cancer. AB - Chemoradiotherapy has become the standard treatment for patients with locally advanced non-small-cell lung cancer on the basis of several large randomized trials. Despite an increase in median survival from 10 months with radiotherapy alone to 16 to 17 months with concurrent chemoradiotherapy, long-term survival in this disease remains modest at best. With the advent of new biologic agents targeting specific cellular pathways associated with malignant progression, combined-modality therapy has the potential to target tumors selectively with less toxicity. PMID- 12781075 TI - Adjuvant therapy in completely resected non-small-cell lung cancer. AB - Less than 20% to 25% of patients with non-small-cell lung cancer (NSCLC) present with stage I or II disease and are best treated by surgical resection. Long-term survival in early NSCLC remains poor. The 5-year survival rate of patients who undergo complete surgical resection is only 40% to 50%. The majority of relapses after surgery are distant metastases; the risk of a local recurrence after complete resection is less than 10%. Postoperative treatments, including chemotherapy, radiotherapy, or both modalities together, have been evaluated widely, but unfortunately none of these treatments have demonstrated any significant impact on survival. Data regarding large-scale adjuvant chemotherapy trials that were closed for accrual almost 4 to 5 years ago will be fully available before the end of the year. It is hoped that a specific meta-analysis will be performed on the basis of these data. PMID- 12781077 TI - Advances in the management of malignant mesothelioma. AB - Malignant mesotheliomas are very aggressive tumors that originate from mesothelial cells, which form the serosal lining of the pleura, pericardial, and peritoneal cavities. Finding effective chemotherapeutic treatment for malignant mesothelioma is a challenge. There is no standard treatment because this tumor is relatively resistant to therapy. A resurgence of interest has been expressed in novel therapies and conventional treatments used in different ways. Several treatment modalities have been studied, including chemotherapy, radiotherapy, surgery, and immunotherapy. Chemotherapy can be administered systemically or directly into the pleura. This review presents the results of the most recent trials and highlights the most promising advances in the battle against this aggressive disease. PMID- 12781076 TI - Targeted therapy for lung cancer. AB - Lung cancer remains the leading cause of death in men and women in the United States. This status is attributed to late diagnosis and lack of effective chemotherapy for metastatic lung cancer. Understanding of the mechanism by which mutated genes confer a neoplastic phenotype on cells has resulted in the development of many potential targeted cancer therapies. This article briefly discusses the work being done with some of these targeted agents in the treatment of lung cancer. PMID- 12781079 TI - [Diagnosis of meningococcal disease by polymerase chain reaction]. AB - BACKGROUND AND OBJECTIVE: The objective of the present study was to evaluate the application of a rapid and simple PCR technique to diagnose meningococcal disease. PATIENTS AND METHOD: A retrospective study was undertaken from January 1999 to June 2002, comprising 110 samples of cerebrospinal fluid (CSF) or plasma from 110 different pediatric patients attending the Hospital Sant Joan de Deu of Barcelona. The selection of patients was based on their diagnosis at discharge: Forty three patients had a discharge diagnose of meningococcal disease (13 meningitis, 12 sepsis and 18 sepsis with meningitis) while 67 had clinical conditions other than meningococcal disease. The samples were processed following standard bacteriological methods (Gram smear and culture) and a PCR technique designed to amplify a segment of the meningococcal insertion sequence IS1106 was performed. RESULTS: Sensitivity of PCR in the group of patients with a clinical diagnosis of meningococcal disease was 93% while sensitivity of the culture was 55.8%. Samples from 19 patients were processed once treatment with -lactam antibiotics had begun (range 8-144 hours), and positive PCR results were seen in 17 cases (sensitivity: 89.4%); a positive culture was observed in two cases of pre-treated patients (sensitivity 10.5%). A false positive result was detected in the group of patients with non-meningococcal disease (specificity 98.5%). CONCLUSIONS: The application of this PCR permits a rapid (roughly 5 hours), specific and sensitive method that increases the microbiologic confirmation of meningococcal disease, mainly in patients who have received previous antibiotic treatment. PMID- 12781078 TI - Treatment options for brain metastases in patients with non-small-cell lung cancer. AB - Brain metastases are a common complication for patients with non-small-cell lung cancer and a significant cause of morbidity and mortality. In the past, treatment of brain metastases and lung cancer focused on symptom palliation with whole brain radiotherapy (WBRT) and steroids because of the grim outlook for patients. However, recent advances in technology and surgical techniques have created more options for the management of brain metastases, which include surgery, irradiation, stereotactic radiosurgery, and chemotherapy. These aggressive approaches have resulted in an improvement of neurologic outcomes and survival rates of patients with non-small-cell lung cancer. Central nervous system (CNS) metastases can be divided into three groups: solitary CNS metastases with controlled or controllable primary disease, oligometastatic disease (fewer than 3 metastases), and multiple metastases. For patients with solitary CNS metastases, long-term survival is possible. A radical treatment approach involving surgical resection or radiosurgery, followed by WBRT, is recommended. For patients with oligometastatic disease, surgical resection or radiosurgery is considered in selected cases and WBRT is indicated. For patients with multiple metastases, WBRT is recommended. For patients with oligometastatic disease and those with multiple metastases, recent evidence indicates that systemically effective chemotherapy may produce responses and can be instituted safely before radiotherapy. The treatment timing of chemotherapy and radiotherapy should be individualized. PMID- 12781080 TI - [Prevalence of infections by hepatitis A, B, C and E viruses in two different socioeconomic groups of children from Santa Cruz, Bolivia]. AB - BACKGROUND AND OBJECTIVES: The epidemiology of hepatitis A, E, B and C was analyzed in 1,393 children living in Santa Cruz de la Sierra, Bolivia. They were distributed in two groups according to the social condition. MATERIALS AND METHOD: 1,393 children were selected from two different schools: one attended by children belonging to a high social class of the town (group A), and the other school attended by children belonging to the poorest social class (group B). Blood samples were drawn by a team of physicians from Rome University La Sapienza. Serum antibodies against hepatitis A, B, C and E virus, and the hepatitis B surface antigen were evaluated by immunometric methods. The significance was evaluated using the *2 test. RESULTS: Antibodies against hepatitis A virus were detected in 82% of examined children, with a significant difference between the two groups (56.3% vs 94.8%). The incidence of anti-HBc antibodies increased with age, so the infection is acquired prevalently in adolescence with a significant difference between both groups (1.1% vs 3.8%). The same phenomenon was observed with anti-HCV antibodies (4.7% positivity only in group B). Serum antibodies against hepatitis E virus were observed in 1.7% cases. CONCLUSIONS: In Bolivia, as in other developing countries, viral hepatitis represents a serious burden for public health. Spreading of viral hepatitis can be controlled upon improving hygienic conditions and customs. Moreover, a vaccination plan against hepatitis A and B virus is necessary for the population living in endemic areas. PMID- 12781081 TI - [Control of arterial hypertension based on self-measurement of blood pressure: APACHE study]. AB - BACKGROUND AND OBJECTIVE: The objective was to analyze the control rates of hypertension in a large group of hypertensives on antihypertensive treatment by means of blood pressure (BP) self-measurements, and to assess if the degree of control at early morning hours differs from that at the rest of the day. PATIENTS AND METHOD: 9,227 subjects [4,398 men, mean age 61.0 (11.4) yr, body mass index 28.4 (4.1) kg/m2] who had hypertension and were on antihypertensive treatment at least during one month before were included. Patients were recruited from primary care physicians. After receiving training, six BP measurements over one day were performed by means of a Braun Precision Sensor BP 2550 monitor, which measures BP at wrist level. RESULTS: Whole day SBP and DBP averages were 138.4 (16.5) mmHg and 85.4 (8.7) mmHg, respectively. Both SBP and DBP in early morning hours were significantly higher than those observed before lunch or before dinner. The percentage of subjects with BP < 130/80 mmHg was 15.5%; 15.4% had SBP < 130 mmHg but DBP > 80 mmHg, and 2.0% had DBP < 80 mmHg and SBP > 130 mmHg). Control rates were lower at early morning hours and in men. CONCLUSIONS: By using BP self measurement, control rates of hypertension are low, especially in early morning hours as compared to the rest of the day. New strategies are needed to further increase the control of hypertension. PMID- 12781082 TI - [Emergency department discharge reports as a management tool of clinical information]. AB - BACKGROUND AND OBJECTIVES: Our goal was to assess the degree of readability, quality and amount of information included in discharge reports of Emergency Departments (ED) in order to evaluate their usefulness for the management of clinical information. MATERIAL AND METHOD: Analysis of a sample of ED discharge reports from 6 Barcelona hospitals (Hospital General Vall d'Hebron, Hospital Clinic de Barcelona, Institut Municipal d'Assistencia Sanitaria [IMAS] hospitals, Hospital Germans Trias i Pujol and Hospital Sant Joan de Deu). We assessed their readability together with the information included for the development of an ED Uniform Discharge Data Set (CMBDAU). RESULTS: 11,188 ED discharge reports were analyzed. Overall, 39% reports were found to have enough and readable information. However, the interpretation of the written information was deemed to be fairly difficult in 46% cases and very difficult in the remaining 15% reports. CONCLUSIONS: Formal quality and comprehensiveness of the information included in ED discharge reports is a critical factor for the precision and usefulness of any registry of data and ED patients classification systems. PMID- 12781083 TI - [Selection of patients for sentinel lymphnode biopsy in melanoma]. PMID- 12781084 TI - [Meningitis caused by streptococci other than S. pneumoniae: clinical and microbiological features of 13 cases]. AB - BACKGROUND AND OBJECTIVE: Meningitis caused by streptococci other than S. pneumoniae are uncommon. We present our experience. PATIENTS AND METHOD: We reviewed the clinical, microbiological, and epidemiological features of all patients with meningitis due to streptococci other than S. pneumoniae diagnosed in the Hospital Universitari Vall d'Hebron during the period 1991-2001. RESULTS: Main causative agents in 13 studied patients were viridans group streptococci (6 cases) and S. agalactiae (5 cases). There were no differences in the clinical presentation between different streptococci. Bacteremia was common in meningitis due to S. agalactiae (80%). Only one patient died. CONCLUSIONS: Meningitis caused by streptococci other than S. pneumoniae are often related to a distant focus of infection or to neurosurgical procedures and, in our experience, they seem to have a good outcome. PMID- 12781085 TI - [Indications of PET imaging]. PMID- 12781086 TI - [Mechanisms and function of sleep: clinical relevance]. PMID- 12781087 TI - [Delayed diagnosis in the traumatic rupture of the pancreas and the pancreatic duct]. PMID- 12781088 TI - [Hipercalcemic crisis after iodine contrast in a patient with parathyroid adenoma]. PMID- 12781089 TI - [Recurrent hypovolemic shock as an onset form of Wunderlich syndrome]. PMID- 12781090 TI - [A work position forever: a golden cage]. PMID- 12781091 TI - [Elective or programmed surgery?]. PMID- 12781092 TI - [Reversible and dose-related parkinsonism induced by venlafaxine]. PMID- 12781093 TI - [Plasma homocysteine levels in systemic lupus erythematosus]. AB - BACKGROUND AND OBJECTIVE: Cardiovascular disease is a major cause of morbidity and mortality in patients with systemic lupus erythematosus (SLE). An association between hyperhomocysteinemia and increased cardiovascular risk has been reported. On the other hand, renal failure and deficiency of vitamin B12 and/or folic acid are common causes of hyperhomocysteinemia. The aims of this study were to determine plasma total homocystein (tHcy) concentrations in SLE patients and to analyze the association of plasma tHcy with age, sex, plasma creatinine, vitamin B12, folates and total cholesterol, as well as with other clinical conditions linked to atherothrombosis in SLE patients. PATIENTS AND METHOD: Fasting plasma levels of tHcy, vitamin B12, folates, total cholesterol and creatinine were measured in 94 SLE patients (11 males, 83 females) and in a control group of 308 healthy volunteers (122 males, 186 females). A review of the medical records of SLE patients was performed. RESULTS: Plasma tHcy concentrations were higher in patients with SLE (median 10.54 (mol/L) than in controls (median 8.49 (mol/L, p < 0.001). Hyperhomocysteinemia (tHcy >=15 (mol/L) was found in 17.02% SLE patients. In a multivariate analysis, plasma creatinine (p < 0.001), total cholesterol (p = 0.038), male sex (p = 0.003) and smoking (p = 0.001) were associated with higher plasma tHcy concentrations. No associations were found between plasma tHcy and hypertension, SLE duration, prednisone therapy and antiphospholipid antibodies. CONCLUSIONS: Plasma tHcy concentrations are higher in SLE patients than in healthy controls. High concentrations of plasma creatinine and total plasma cholesterol, male sex and smoking are associated with a higher concentration of plasma tHcy in SLE. Since the clinical consequences of hyperhomocysteinemia are not well established, routine determination of plasmatic tHcy and supplemental therapy in patients with high levels of tHcy are not recommended. PMID- 12781095 TI - [A comparative psychometric study of the Spanish versions with 6, 17, and 21 items of the Hamilton Depression Rating Scale]. AB - BACKGROUND AND OBJECTIVE: Former Spanish studies on the Hamilton Depression Rating Scale (HDRS) have focused on the 17 items version and have been conducted in depressive in-patient series. This study compares the psychometric properties of the Spanish version of HDRS with 6, 17 and 21 items as assessed in depressive patients in ambulatory care. PATIENTS AND METHOD: A prospective, observational and multicenter study was conducted in clinically stable or unstable depressed patients. The discriminative validity, reliability (internal consistency, test retest and inter-raters) and sensitivity to change were assessed. RESULTS: One hundred and sixty eight patients from 15 psychiatric facilities were included. The 6, 17 and 21 items versions of HDRS showed appropriate discriminative validity (HDRS-Clinical Global Impression: p < 0.0001); internal consistency (Cronbach *: HDRS-6 = 0.6; HDRS-17 and 21 >= 0.7); test-retest reliability (intraclass correlation coefficient [ICC]: HDRS-6, 17 and 21 >= 0.9); inter raters reliability (ICC: HDRS-6, 17 and 21 >= 0.9) and sensitivity to change (effect size >= 1.5 for HDRS-6, 17 and 21). CONCLUSIONS: Spanish versions of HDRS with 6, 17 and 21 items show similar psychometric properties. HDRS-6 could be useful in ambulatory and primary care. PMID- 12781094 TI - [Administration-time dependent effects of acetyl-salicylic acid on blood pressure in patients with mild essential hypertension]. AB - BACKGROUND AND OBJECTIVES: Previous studies on the potential influence of aspirin (AAS) on blood pressure have not taken in consideration the chronopharmacologic effects of non-steroidal antiinflammatory drugs. This pilot study investigated the effects of AAS on blood pressure in hypertensive patients under no antihypertensive treatment who received AAS at different day times according to their rest-activity cycle. PATIENTS AND METHOD: We studied 64 untreated patients with mild hypertension (24 men), 43.5 (12.0) (mean [SD]) years of age, randomly divided in 3 groups: group 1, non-pharmacological hygienic-dietetic recommendations; group 2, the same recommendations plus AAS (100 mg/day) on awakening, and group 3, the same recommendations plus AAS (100 mg/day) before bedtime. Blood pressure was measured every 20 min during the day and every 30 min at night for 48 consecutive hours before and after 3 months of intervention. The circadian pattern of blood pressure in each group was established by population multiple-component analysis. Circadian parameters obtained for each group of patients before and after the intervention were compared with a nonparametric paired test. RESULTS: After 3 months of non-pharmacological intervention, there was a small and non-significant reduction of blood pressure (< 1.5 mmHg; p = 0.28). There was no change in blood pressure when AAS was given on awakening (p = 0.21). A highly significant blood pressure reduction was, however, observed in patients who received AAS before bedtime (decrease of 7 and 5 mmHg in systolic and diastolic blood pressure, respectively; p = 0.006). CONCLUSIONS: Our results indicate a statistically significant time-dependent administration effect of low dose AAS on blood pressure in untreated patients with mild hypertension. This effect reinforces the need to quantify and control AAS effects in patients using this agent in combination with antihypertensive agents. PMID- 12781096 TI - [Assessment of adherence to antiretroviral: role of determination of plasma levels of non nucleosides anologues]. AB - BACKGROUND AND OBJECTIVE: Our purpose was to assess the contribution of efavirenz and nevirapine plasma concentrations in the assessment of self-reported adherence and punctuality of appointments to take the drugs. PATIENTS AND METHOD: Adherence was assessed using questionnaires and plasma concentrations of efavirenz and nevirapine were determined. Patients received efavirenz or nevirapine in combination with two nucleotides reverse transcriptase inhibitors. RESULTS: Among the 45 enrolled patients, 7 were considered non-adherent according to the questionnaires, 17 regarding pharmacy appointments and 5 according to the plasma levels of efavirenz and nevirapine. 3.3% participants had detectable viral loads. CONCLUSION: Cross-validation between patients'self-reports and pharmacy appointments can improve the measurement of adherence to antiretroviral therapy. PMID- 12781097 TI - [Iron overload. More than hereditary hemochromatosis]. PMID- 12781098 TI - [The future of scientific training in the hospitals of the Spanish Health System]. PMID- 12781099 TI - [Hospital emergency rooms]. AB - Overuse of hospital emergency rooms (HERs) is parallel to their controversy. To understand this problem, some concepts should be first clarified. In HERs, there are some intrinsic aspects which are directly related to the emergency itself and thus cannot be modified (intermittent patient flow, need to prioritize, difficulty to achieve a rapid diagnosis, influence of time on treatment, value of clinical follow up, patient's expectations, impact of HER on the overall hospital working dynamics). On the other hand, there are some extrinsic aspects which indeed are not related to HER itself but are rather historically associated with it (precarious structure, delay on admission, lack of privacy, inadequate triage of cases, lack of professionalization); these latter aspects may be potentially modified and should be reconsidered. PMID- 12781102 TI - [Crossed bucofacial apraxia]. PMID- 12781100 TI - [Is it necessary to reduce the LDL cholesterol level below 100 mg/dl in all diabetic patients?]. PMID- 12781103 TI - [False-positive serological tests in malaria]. PMID- 12781104 TI - [Adrenal masses in Gardners syndrome: report of two cases]. PMID- 12781105 TI - [Drug abuse in pregnancy and neonatal morbidity]. PMID- 12781106 TI - [Overweight and risk of overweight in schoolchildren aged 6-17 years old in Galicia (Spain)]. AB - BACKGROUND: Overweight and obesity currently represent one of the most important public health problems in industrialized countries and are also linked to cardiovascular disease and sleep apnea, as well as to orthopaedic and other complications. OBJECTIVES: To examine overweight and the risk of overweight in the school-aged population in Galicia in comparison with the nationally representative survey in the US population: the first National Health and Nutrition Examination Survey (NHANES-1). METHODS: We performed a cross sectional study including a sample of 1105 boys and girls (50 % from a rural environment and 50 % from urban areas) in Galicia, northwest Spain. RESULTS: The 50th percentile for boys and girls in Galicia was higher than the corresponding percentile in the NHANES-1 for all the ages studied. In general, the 85th and 95th percentiles for children's body mass index in Galicia were higher in the group aged 6-14 years. CONCLUSIONS: In Galicia body mass index values in the 50th, 85th and 95th percentiles were higher in girls than in boys. For the 50th percentile, both boys and girls in Galicia had higher values than those of the NHANES-1 for all ages. In areas with risk of excess weight (85th percentile) and prevailing excess weight (95th percentile) the same was true in the 6 to 13-14 age group for both sexes. PMID- 12781107 TI - [Growth in children with juvenile idiopathic arthritis]. AB - INTRODUCTION: Juvenile idiopathic arthritis (JIA) is a chronic inflammatory disease that often requires steroid therapy. Growth retardation can be a serious complication in some of these patients. OBJECTIVE: To study linear growth in patients with JIA and evaluate the factors involved in its disturbance. METHODS: We studied 91 patients with JIA with a follow-up of at least one year. A cross sectional study, a longitudinal retrospective study, and a longitudinal prospective study were performed. Height in the first consultation, in the cross sectional study, and one year previously was evaluated. Height velocity (HV) was calculated. Several parameters related to disease activity, corticosteroid therapy, nutritional and hormonal status, and bone mineral density (BMD) were analyzed. A correlation study and multivariate regression analysis were carried out. RESULTS: Height was < or = -2 SD in 14.3 % of the series and in 55.6 % of the systemic group. Variables independently associated with height were total corticosteroid dose, functional class, nutritional index, BMD, and age at onset of the disease. HV was < or = 2 SD in 25.3 % of the series and in 61 % of the systemic group. Variables independently associated with HV were corticosteroid dose and the number of swollen joints. CONCLUSIONS: Growth retardation in patients with JIA was associated with factors related to the disease, corticosteroid therapy, nutritional status, BMD, and earlier onset of the disease. PMID- 12781108 TI - [Determination of nicotine dependence in school-aged smokers through a modified Fagerstrom Test]. AB - BACKGROUND: Approximately 65 % of young smokers have failed to give up smoking because of their addiction to nicotine. The Fagerstrom Test for Nicotine Dependence is used to quantify nicotine dependence in adults but studies in teenagers are scarce. OBJECTIVES: To determine whether young smokers are nicotine dependent, and if so, to what extent, as well as the factors linked to nicotine dependence. METHODS: A total of 2647 schoolchildren, aged 10 to 17 years old, from 41 different schools were interviewed. A sample size was calculated for each age, in years, by using the equation of finite population, but adding an extra 10 % so that if the numbers decreased, the final absolute error would not be increased. For each age the sample was stratified by sex and type of school, with proportional affixation to the number of individuals within each stratum. Schools and students were chosen by using the random numbers table. In addition to eliciting personal data, the questionnaire also included a version of the FTND adapted for use in teenagers. RESULTS: The mean age was 13.77 years and 51.5 % were male. A total of 23.1 % reported they were smokers (54.5 % of them were girls and 45.5 % were boys). Most (86.6 %) of these teenager smokers showed low moderate nicotine dependence and 3.3 % showed high dependence. No differences were found according to sex or age. CONCLUSIONS: Most teenagers who smoke show low nicotine dependence. Campaigns to prevent smoking should be aimed at children aged less than 10 years old to delay smoking the first cigarette and the subsequent next step of becoming a daily smoker for as long as possible since both factors have proved decisive in nicotine dependence. PMID- 12781109 TI - [Ambulatory blood pressure monitoring in multicystic dysplastic kidney disease]. AB - OBJECTIVE: To evaluate blood pressure in a group of patients with multicystic dysplastic kidney (MDK) disease through ambulatory blood pressure (ABP) monitoring. PATIENTS AND METHODS: ABP monitoring was performed in 16 patients with MDK disease using a Spacelabs 90207 monitor and an appropriately sized armband. The patients performed their normal daily activities except physical exercise. The activity period was 8:00 am to 8:00 pm and the resting period was midnight to 6:00 am. Patients with a percentage of correct readings of less than 70 % were excluded. The mean systolic and diastolic blood pressures for each period were calculated. Circadian variability was determined by the ratio between mean systolic and diastolic values in the active and resting periods, respectively. The results were compared with ABP values in healthy children. RESULTS: Fourteen patients (mean age 16 3.1 years, range: 6-27.6) were included. No patients with hypertension were found through casual measurement. One patient had daytime systolic hypertension. Five patients presented an abnormal drop in blood pressure during sleep. CONCLUSION: Blood pressure should be monitored in patients with MDK disease. ABP monitoring may detect alterations that can go unnoticed in casual determinations. Further studies with large samples are needed to establish the real prevalence of hypertension in these patients. PMID- 12781110 TI - [Quality of life evaluation in children and adolescents with chronic and/or incapacitating diseases: a Brazilian study]. AB - OBJECTIVE: To evaluate quality of life in children and adolescents with acute lymphocytic leukemia (ALL) and juvenile rheumatoid arthritis (JRA). MATERIAL AND METHODS: We administered the Children's Global Assessment Scale (CGAS), the Vineland Adaptative Behavior Scale (VABS) and the Autoquestionnaire qualite de vie enfant image (AUQEI) to a sample of 28 children with ALL, 28 children with JRA, and 28 healthy controls, aged 4 to 13 years old, who were diagnosed between 1 and 5 years previously. RESULTS: Slight differences were found in age between patients with ALL and those with JRA. No significant differences were found in time since diagnosis or in CGAS scores. A significant difference was found in VABS global scores, as well as in VABS communication domain scores. No significant differences were found in VABS daily living skills domain scores between patients with ARJ and healthy controls. No significant differences were found among the groups in VABS socialization domain scores or in AUQEI scores. CONCLUSION: In our study, chronically ill children clearly performed worse in adaptative behavior development. Nevertheless, their quality of life was similar to that of healthy controls. Appropriate methods to identify pediatric patients' perception of their illnesses and treatment should be urgently developed. PMID- 12781111 TI - [Diagnostic yield of abdominal ultrasonography in acute appendicitis]. AB - INTRODUCTION: Acute appendicitis (AA) is the first cause of surgical abdomen in childhood but diagnosis continues to be difficult in some cases. Abdominal ultrasonography has been shown to be useful when there is doubt about the diagnosis and can be quickly and easily performed at the patient's bedside. OBJECTIVE: To evaluate the efficacy of abdominal ultrasonography in the diagnosis of acute appendicitis in our hospital. PATIENTS AND METHODS: All patients who visited the emergency department for abdominal pain and who underwent abdominal ultrasonography to rule out AA between January 1, 1999 and December 31, 2000 were retrospectively studied. RESULTS: During the study period, 4217 children were evaluated in our service for abdominal pain. Ultrasonography was performed in 528 children. Of these, the procedure was performed to rule out AA in 308 patients. Of these 308 patients who met the study's inclusion criteria, the results of ultrasonography were compatible with AA in 102 and were normal in 196. Ultrasonographic diagnosis differed from the final diagnosis in 16 patients. In eight children with ultrasonographic findings suggestive of AA, laparotomy revealed normal appendices. In the remaining eight patients, ultrasonographic findings were normal and surgery revealed AA. Based on these findings, the diagnostic yield of abdominal ultrasonography was: sensitivity: 96.6 %; specificity: 95.9 %, positive predictive value: 86 %; negative predictive value: 95.9 %. CONCLUSIONS: The overall diagnostic yield of abdominal ultrasonography in AA our hospital is acceptable. Because ultrasonographic diagnosis is not always accurate, others diagnostic methods such as computed tomography should be used in doubtful cases. PMID- 12781112 TI - [Fetal-neonatal alloimmune thrombocytopenia]. AB - Fetal-neonatal alloimmune thrombocytopenia is the commonest cause of severe thrombocytopenia in the newborn. This disorder is due to the destruction of fetal platelets by a maternal platelet-specific antibody caused by fetal-maternal incompatibility. The most serious complication is intracranial hemorrhage (10-30 % of newborns), which may cause death (10 % of the reported cases) or irreversible neurological sequelae (20 %). The diagnosis is usually made after birth when most affected neonates have petechiae, purpura or overt bleeding. The degree of severity varies according to platelet count. Current methods allow detection of maternal platelet alloantibodies (usually HPA-1a). Clinical grounds and the exclusion of other causes of neonatal thrombocytopenia are required to establish an accurate diagnosis. Recurrence of this disease is very high and has prompted clinicians to develop antenatal prophylactic programs in subsequent pregnancies. However, the optimal treatment of at-risk pregnancies remains controversial. The early diagnosis of this process allows effective therapy based on the infusion of compatible platelets and IgG immunoglobulins when hemorrhage is not obvious. Antenatal management of subsequent pregnancies can prevent recurrence of thrombocytopenia and intracranial hemorrhage. The aim of this review is to draw pediatricians' attention to the importance of this probably under-diagnosed disease in which early diagnosis can prevent potentially severe complications. PMID- 12781113 TI - [Dietary changes in Mexico]. AB - Although the Mexican population has traditionally been malnourished, the prevalence of obesity in children and adults has increased by almost 50 % in the last 10 years. Recent studies show substantial changes in the nutritional status of Mexicans, especially in the pediatric population. Among the factors associated with the development of obesity are overeating, sedentariness, and genetics. The apparent economic development in Mexico, as well as the influence of dietary patterns from other countries, have contributed to modifying lifestyle. Despite measures taken by the health system, iron- and zinc-deficiency anemia continue to be prevalent. The present review aims to describe the changes that have taken place in Mexico in the last few decades leading to a generation of short and obese children, as well as to determine the associated factors in order to promote healthier eating patterns among the Mexican population. PMID- 12781114 TI - [Drug abuse in pregnancy and neonatal morbidity: epidemiologic changes in the last ten years]. AB - BACKGROUND: In the last few years, patterns of drug abuse have varied. This may have changed the problems presented by the infants of drug-abusing mothers. OBJECTIVES: To determine the incidence of prenatal drug-exposure in neonates in our health area in the last decade, to analyze possible changes in the drugs consumed, routes of administration and the infectious diseases affecting drug abusing mothers, and to study possible changes in the clinical features presented by their offspring. PATIENTS AND METHODS: An observational, descriptive, retrospective study of all the infants born to drug-abusing mothers from January 1, 1992 to December 31, 2001 was performed. The series was analyzed and the periods 1992-1996 and 1997-2001 were compared. RESULTS: The mean incidence was 7.5 cases per 1000 live-born infants (8.7 in 1992-1996 and 6.3 in 1997-2001). Mothers in the second half of the study were less likely to use heroin and the intravenous route and were more likely to use methadone and cocaine than mothers in the 1997-2001 period. Hepatitis C virus infection and instrumental delivery were more frequent in the 1997-2001 period. No differences were found in neonates born in either period. Birthweight was less than 2500 g in 40 % and was low for gestational age in 20 %. Preterm deliveries were performed in 24 %. Congenital malformations were found in 6 %, neonatal withdrawal syndrome in 79 %, human immunodeficiency virus infection in 2 %, and hepatitic C virus infection in 3 %. Four percent were given in adoption. CONCLUSIONS: Maternal drug abuse patterns changed in the ten years under study, but outcome in their offspring showed no differences. Drug abuse by pregnant women is linked to a high rate of low birthweight, prematurity, neonatal withdrawal syndrome, vertical infection by human immunodeficiency virus and hepatitis C virus, as well as to social and family problems. PMID- 12781115 TI - [Arteriovenous malformations of the vein of Galen]. AB - Vein of Galen arteriovenous malformations encompass a diverse group of vascular anomalies that share a common feature: dilatation of the vein of Galen. Although clinical presentations are highly variable, depending on age of presentation, signs and symptoms overlap between age groups. The association of heart failure and cranial bruit constitutes the most striking clinical presentation in neonates. However, less severe and fulminant modes of presentation are frequent in older infants, children, and adults. Treatment approaches consist of symptomatic treatment of heart failure on the one hand and of surgery or endovascular treatment on the other. The results of the latter have improved in recent years, opening up a broad spectrum of new possibilities. We present the case of an asymptomatic 15-day-old neonate who presented an arteriovenous malformation of the vein of Galen and who was treated with endovascular occlusion of the arterial afferents. An excellent result was obtained with no evidence of neurological abnormalities. PMID- 12781116 TI - [Consumption of fruit juices and beverages by Spanish children and teenagers: health implications of their poor use and abuse]. AB - In the last few decades, the consumption of soft drinks (fruit juices and beverages) in Spain has increased considerably. From 1991 to 2001, consumption of soft drinks increased by 41.5 %, that of extract-based drinks by 62.1 % and that of juices by 26.7 %. Consumption increases with age, with teenagers drinking twice as much (740 ml/day) as pre-school children (388 ml/day). High consumption of soft drinks may lead to underconsumption of foods and drinks of high nutritional quality, such as milk. Multiple studies relate this phenomenon with the risk of growth retardation, failure of non-organic growth, diarrhea produced by alterations in carbohydrate absorption, allergy, pharmacological interactions, obesity, atherothrombotic lipidic profile, alterations in glucose and bone mineral metabolism and negative effects on dental health. Because excessive consumption of soft drinks favors a diet of low nutritional quality, intervention and prevention strategies should be established to promote water and milk as the basic drink in children's and teenagers' diets while soft-drinks should be consumed only occasionally. PMID- 12781117 TI - [Subacute sclerosing panencephalitis: a still existing disease]. AB - Subacute sclerosing panencephalitis (SSPE) is a rare entity with an invariably fatal course that progressively affects the central nervous system. It is caused by persistent infection with the wild-type measles virus. While rare in industrial countries, it is not infrequent in developing countries, where there are still areas of endemic measles infection and immunization is not yet generalized. We describe an eight-year-old Spanish girl who presented rhythmic and symmetric myoclonus. She contracted measles at 13 months and required hospitalization. No cognitive deterioration was found. Neuroimaging and the initial electroencephalogram were normal. Oligoclonal bands and high titers of measles antibodies were found in serum and cerebrospinal fluid. She was treated with oral metisoprinol and intraventricular alpha-interferon (IFN-) and showed no further progression of her symptoms. The importance of including SSPE in the differential diagnosis of patients consulting for school failure, neurological deterioration or movement disorders is highlighted. Special attention should be paid to the immigrant population from countries where the incidence of SSPE is greater than in Spain. PMID- 12781118 TI - [Heterogenicity of Gorham-Stout syndrome: association with lymphatic and venous malformations]. AB - INTRODUCTION: Gorham-Stout syndrome is a rare disorder of unknown etiology characterized by osteolysis and microscopic proliferation of abnormal vessels. We report two cases of this syndrome associated with lymphatic and venous malformations. CASE REPORTS: The first case is a 5-year-old boy with disseminated lymphangiomatosis of poor prognosis, with significant pleural involvement and osteolytic lesions. The second case is a 5-year-old girl with a diagnosis of Klippel-Trenaunay syndrome with significant skeletal involvement of the lower extremities and secondary pathological fracture. CONCLUSIONS: Gorham-Stout syndrome may occasionally be associated with various lymphatic and venous malformations. Osteolysis and bone resorption may be induced by lymphatic bone involvement. PMID- 12781119 TI - [Giant pleural fibroma]. AB - Solitary fibrous tumors of the pleura are uncommon mesenchymal tumors that have been reported in adults. There are very few cases in children. These tumors are considered benign but their behavior is often unpredictable because they can invade nearby structures and transform into malignant variants after several years. Their clinical presentation is diverse but generally asymptomatic and they are detected incidentally in routine radiologic studies. Prognosis depends firstly on resectability of the tumour and secondly on its size, mitotic count, polymorphism, and necrosis within the tumour. Surgical resection is usually curative. Long-term follow-up is recommended because local recurrences can occur after surgery. We report the case of an asymptomatic 15-year-old girl with a giant thoracic mass revealed incidentally on a chest radiograph. Physical examination revealed thoracic asymmetry and no sounds of breathing were found in the right lung. Computerized tomography and magnetic resonance images showed the giant size of the mass. Preoperative fine needle biopsy was performed. Intraoperative biopsy confirmed the diagnosis. Although histologically benign, the tumors may enlarge rapidly. In the present case, because of the localization and size of the tumor, the patient was at increased risk. The tumour was resected and the postoperative course was uneventful. At present our patient remains under long term annual follow-up. PMID- 12781121 TI - [Isolated obstetric costal fractures]. PMID- 12781120 TI - [Familial hypobetalipoproteinemia]. AB - We present the case of a 5-month-old girl, with consanguineous parents, who was born at 28 weeks of gestation and who showed intermittent signs of abdominal distension accompanied by increased regurgitation and vomiting after food intake. Significant biochemical alterations (reduced levels of triglicerides, cholesterol, and vitamin A and absence of apolipoprotein B and vitamin E) led to the diagnosis of homozygous hypobetalipoproteinemia, which was subsequently confirmed by genetic studies. PMID- 12781122 TI - [Discitis and spondylodiskitis in young children: Difficulties in making an early diagnosis]. PMID- 12781123 TI - [Urinary tract infection caused by Shigella sonnei]. PMID- 12781124 TI - [Lymphomatoid papulosis]. PMID- 12781125 TI - [Liposuction as palliative treatment of giant lipoma]. PMID- 12781126 TI - [Rapid development of pneumonia]. PMID- 12781127 TI - The ubiquitin proteasome system functions as an inhibitory constraint on synaptic strengthening. AB - BACKGROUND: Long-lasting forms of synaptic plasticity have been shown to depend on changes in gene expression. Although many studies have focused on the regulation of transcription and translation during learning-related synaptic plasticity, regulated protein degradation provides another common means of altering the macromolecular composition of cells. RESULTS: We have investigated the role of the ubiquitin proteasome system in long-lasting forms of learning related plasticity in Aplysia sensory-motor synapses. We find that inhibition of the proteasome produces a long-lasting (24 hr) increase in synaptic strength between sensory and motor neurons and that it dramatically enhances serotonin induced long-term facilitation. The increase in synaptic strength produced by proteasome inhibitors is dependent on translation but not transcription. In addition to the increase in synaptic strength, proteasome inhibition leads to an increase in the number of synaptic contacts formed between the sensory and motor neurons. Blockade of the proteasome in isolated postsynaptic motor neurons produces an increase in the glutamate-evoked postsynaptic potential, and blockade of the proteasome in the isolated presynaptic sensory cells produces increases in neurite length and branching. CONCLUSIONS: We conclude that both pre- and postsynaptic substrates of the ubiquitin proteasome function constitutively to regulate synaptic strength and growth and that the ubiquitin proteasome pathway functions in mature neurons as an inhibitory constraint on synaptic strengthening. PMID- 12781128 TI - The ubiquitin proteasome system acutely regulates presynaptic protein turnover and synaptic efficacy. AB - BACKGROUND: The ubiquitin proteasome system (UPS) mediates regulated protein degradation and provides a mechanism for closely controlling protein abundance in spatially restricted domains within cells. We hypothesized that the UPS may acutely determine the local concentration of key regulatory proteins at neuronal synapses as a means for locally modulating synaptic efficacy and the strength of neurotransmission communication. RESULTS: We investigated this hypothesis at the Drosophila neuromuscular synapse by using an array of genetic and pharmacological tools. This study demonstrates that UPS components are present in presynaptic boutons and that the UPS functions locally in the presynaptic compartment to rapidly eliminate a conditional transgenic reporter of proteasome activity. We assayed a panel of synaptic proteins to determine whether the UPS acutely regulates the local abundance of native synaptic targets. Both acute pharmacological inhibition of the proteasome (<1 hr) and targeted genetic perturbation of proteasome function in the presynaptic neuron cause the specific accumulation of the essential synaptic vesicle-priming protein DUNC-13. Most importantly, acute pharmacological inhibition of the proteasome (<1 hr) causes a rapid strengthening of neurotransmission (an approximately 50% increase in evoked amplitude) because of increased presynaptic efficacy. The proteasome-dependent regulation of presynaptic protein abundance, both of the exogenous reporter and native DUNC-13, and the modulation of presynaptic neurotransmitter release occur on an intermediate, rapid (tens of minutes) timescale. CONCLUSIONS: Taken together, these studies demonstrate that the UPS functions locally within synaptic boutons to acutely control levels of presynaptic protein and that the rate of UPS-dependent protein degradation is a primary determinant of neurotransmission strength. PMID- 12781129 TI - Neddylation and deneddylation of CUL-3 is required to target MEI-1/Katanin for degradation at the meiosis-to-mitosis transition in C. elegans. AB - BACKGROUND: SCF (Skp1-Cullin-F-box) complexes are a major class of E3 ligases that are required to selectively target substrates for ubiquitin-dependent degradation by the 26S proteasome. Conjugation of the ubiquitin-like protein Nedd8 to the cullin subunit (neddylation) positively regulates activity of SCF complexes, most likely by increasing their affinity for the E2 conjugated to ubiquitin. The Nedd8 conjugation pathway is required in C. elegans embryos for the ubiquitin-mediated degradation of the microtubule-severing protein MEI 1/Katanin at the meiosis-to-mitosis transition. Genetic experiments suggest that this pathway controls the activity of a CUL-3-based E3 ligase. Counteracting the Nedd8 pathway, the COP9/signalosome has been shown to promote deneddylation of the cullin subunit. However, little is known about the role of neddylation and deneddylation for E3 ligase activity in vivo. RESULTS: Here, we identified and characterized the COP9/signalosome in C. elegans and showed that it promotes deneddylation of CUL-3, a critical target of the Nedd8 conjugation pathway. As in other species, the C. elegans signalosome is a macromolecular complex containing at least six subunits that localizes in the nucleus and the cytoplasm. Reducing COP9/signalosome function by RNAi results in a failure to degrade MEI-1, leading to severe defects in microtubule-dependent processes during the first mitotic division. Intriguingly, reducing COP9/signalosome function suppresses a partial defect in the neddylation pathway; this suppression suggests that deneddylation and neddylation antagonize each other. CONCLUSIONS: We conclude that both neddylation and deneddylation of CUL-3 is required for MEI-1 degradation and propose that cycles of CUL-3 neddylation and deneddylation are necessary for its ligase activity in vivo. PMID- 12781130 TI - C. elegans PAT-6/actopaxin plays a critical role in the assembly of integrin adhesion complexes in vivo. AB - BACKGROUND: The novel focal adhesion protein actopaxin includes tandem unconventional calponin homology (CH) domains and a less well-conserved N terminal stretch. Dominant-negative studies have implicated actopaxin in focal adhesion formation. RESULTS: PAT-6/actopaxin, the sole actopaxin homolog in C. elegans, is located in body wall muscle attachments that are in vivo homologs of focal adhesions. We show using pat-6 protein null alleles that PAT-6/actopaxin has critical nonredundant roles during attachment maturation. It is required to recruit UNC-89 and myofilaments to newly forming attachments, and also to reposition the attachments so that they form the highly ordered array of dense body and M line attachments that are characteristic of mature muscle cells. PAT 6/actopaxin is not required for the deposition of UNC-52/perlecan in the basal lamina, nor for the initiation of attachment assembly, including the clustering of integrin into foci and the recruitment of attachment proteins PAT-4/ILK, UNC 112, and DEB-1/vinculin from the cytosol. PAT-6/actopaxin, PAT-4/ILK, and UNC-112 are each required for the same steps during attachment assembly in vivo, consistent with the notion that they work together in multiprotein complex. Supporting this idea, PAT-4/ILK can simultaneously bind to PAT-6/actopaxin and UNC-112, forming a ternary complex, in yeast three-hybrid assays. Finally, we show that both calponin homology domains are required for PAT-6/actopaxin's critical functions during attachment assembly in vivo. CONCLUSIONS: We show directly by loss-of-function genetics that PAT-6/actopaxin plays essential roles during the maturation of integrin-mediated muscle attachments in vivo. PMID- 12781131 TI - Structure of the Y14-Magoh core of the exon junction complex. AB - BACKGROUND: Splicing of pre-mRNA in eukaryotes imprints the resulting mRNA with a specific multiprotein complex, the exon-exon junction complex (EJC), at the sites of intron removal. The proteins of the EJC, Y14, Magoh, Aly/REF, RNPS1, Srm160, and Upf3, play critical roles in postsplicing processing, including nuclear export and cytoplasmic localization of the mRNA, and the nonsense-mediated mRNA decay (NMD) surveillance process. Y14 and Magoh are of particular interest because they remain associated with the mRNA in the same position after its export to the cytoplasm and require translation of the mRNA for removal. This tenacious, persistent, splicing-dependent, yet RNA sequence-independent, association suggests an important signaling function and must require distinct structural features for these proteins. RESULTS: We describe the high-resolution structure and biochemical properties of the highly conserved human Y14 and Magoh proteins. Magoh has an unusual structure comprised of an extremely flat, six stranded anti-parallel beta sheet packed against two helices. Surprisingly, Magoh binds with high affinity to the RNP motif RNA binding domain (RBD) of Y14 and completely masks its RNA binding surface. CONCLUSIONS: The structure and properties of the Y14-Magoh complex suggest how the pre-mRNA splicing machinery might control the formation of a stable EJC-mRNA complex at splice junctions. PMID- 12781132 TI - Human POT1 facilitates telomere elongation by telomerase. AB - Mammalian telomeric DNA is mostly composed of double-stranded 5'-TTAGGG-3' repeats and ends with a single-stranded 3' overhang. Telomeric proteins stabilize the telomere by protecting the overhang from degradation or by remodeling the telomere into a T loop structure. Telomerase is a ribonucleoprotein that synthesizes new telomeric DNA. In budding yeast, other proteins, such as Cdc13p, that may help maintain the telomere end by regulating the recruitment or local activity of telomerase have been identified. Pot1 is a single-stranded telomeric DNA binding protein first identified in fission yeast, where it was shown to protect telomeres from degradation [10]. Human POT1 (hPOT1) protein is known to bind specifically to the G-rich telomere strand. We now show that hPOT1 can act as a telomerase-dependent, positive regulator of telomere length. Three splice variants of hPOT1 were overexpressed in a telomerase-positive human cell line. All three variants lengthened telomeres, and splice variant 1 was the most effective. hPOT1 was unable to lengthen the telomeres of telomerase-negative cells unless telomerase activity was induced. These data suggest that a normal function of hPOT1 is to facilitate telomere elongation by telomerase. PMID- 12781133 TI - Heterotrimeric G proteins regulate daughter cell size asymmetry in Drosophila neuroblast divisions. AB - Cell division often generates unequally sized daughter cells by off-center cleavages, which are due to either displacement of mitotic spindles or their asymmetry. Drosophila neuroblasts predominantly use the latter mechanism to divide into a large apical neuroblast and a small basal ganglion mother cell (GMC), where the neural fate determinants segregate. Apically localized components regulate both the spindle asymmetry and the localization of the determinants. Here, we show that asymmetric spindle formation depends on signaling mediated by the G beta subunit of heterotrimeric G proteins. G beta 13F distributes throughout the neuroblast cortex. Its lack induces a large symmetric spindle and causes division into nearly equal-sized cells with normal segregation of the determinants. In contrast, elevated G beta 13F activity generates a small spindle, suggesting that this factor suppresses spindle development. Depletion of the apical components also results in the formation of a small symmetric spindle at metaphase. Therefore, the apical components and G beta 13F affect the mitotic spindle shape oppositely. We propose that differential activation of G beta signaling biases spindle development within neuroblasts and thereby causes asymmetric spindles. Furthermore, the multiple equal cleavages of G beta mutant neuroblasts accompany neural defects; this finding suggests indispensable roles of eccentric division in assuring the stem cell properties of neuroblasts. PMID- 12781134 TI - Sensitive flow cytometric analysis reveals a novel type of parent-of-origin effect in the mouse genome. AB - The discovery of classic parental imprinting came, at least in part, from the analysis of transgene expression in mice. It was noticed that some transgenes were only expressed following paternal transmission and that others sometimes showed differential patterns of methylation depending on the parent of origin. Here, we present evidence of a novel and more subtle form of parental imprinting by taking advantage of the highly sensitive detection of murine transgene expression afforded by flow cytometry. We have produced nine lines of transgenic mice carrying a GFP reporter linked to the human alpha-globin promoter and enhancer elements, which direct expression to erythroid cells. A high proportion of transgenic lines, four of the nine, display significantly lower levels of expression following maternal transmission. Both the percentage of expressing cells and the mean fluorescence in expressing cells are between 10% and 30% lower following maternal transmission. These effects are reversible upon passage through the opposite germline. This finding raises the possibility that differences in the epigenetic state of the maternal and paternal chromosomes in adult somatic cells are more widespread than was previously thought. PMID- 12781135 TI - A role of Dishevelled in relocating Axin to the plasma membrane during wingless signaling. AB - Wnt signaling causes changes in gene transcription that are pivotal for normal and malignant development. A key effector of the canonical Wnt pathway is beta catenin, or Drosophila Armadillo. In the absence of Wnt ligand, beta-catenin is phosphorylated by the Axin complex, which earmarks it for rapid degradation by the ubiquitin system. Axin acts as a scaffold in this complex, to assemble beta catenin substrate and kinases (casein kinase I [CKI] and glycogen synthase kinase 3 beta [GSK3]). The Adenomatous polyposis coli (APC) tumor suppressor also binds to the Axin complex, thereby promoting the degradation of beta-catenin. In Wnt signaling, this complex is inhibited; as a consequence, beta-catenin accumulates and binds to TCF proteins to stimulate the transcription of Wnt target genes. Wnt induced inhibition of the Axin complex depends on Dishevelled (Dsh), a cytoplasmic protein that can bind to Axin, but the mechanism of this inhibition is not understood. Here, we show that Wingless signaling causes a striking relocation of Drosophila Axin from the cytoplasm to the plasma membrane. This relocation depends on Dsh. It may permit the subsequent inactivation of the Axin complex by Wingless signaling. PMID- 12781136 TI - Notch steers Drosophila ISNb motor axons by regulating the Abl signaling pathway. AB - The central problem in axon guidance is to understand how guidance signals interact to determine where an axon will grow. Here we investigate a specific axon guidance decision in Drosophila embryos, the sharp inward turn taken by the ISNb motor nerve to approach its muscle targets. We find that this turn requires Notch and its ligand Delta. We show that Delta is expressed on cells adjacent to the ISNb turning point, and we know from previous work that Notch is present on axonal growth cones, suggesting that Delta and Notch might provide a guidance signal to ISNb. To induce the turning of ISNb axons, Notch interacts genetically with multiple components of a signal transduction pathway that includes the Abl tyrosine kinase and its affiliated accessory proteins. In contrast, genetic interaction experiments fail to provide evidence for a major role of the "canonical" Notch/Su(H) signaling pathway in this process. We suggest that the Notch/Abl interaction promotes the turning of ISNb axons by attenuating the Abl dependent adhesion of ISNb axons to their substratum, thus releasing the axons to respond to attraction from target muscles. PMID- 12781137 TI - Importance of a single base pair for discrimination between intron-containing and intronless alleles by endonuclease I-BmoI. AB - Homing endonucleases initiate mobility of their host group I introns by binding to and cleaving lengthy recognition sequences that are typically centered on the intron insertion site (IS) of intronless alleles. Because the intron interrupts the endonucleases' recognition sequence, intron-containing alleles are immune to cleavage by their own endonuclease. I-TevI and I-BmoI are related GIY-YIG endonucleases that bind a homologous stretch of thymidylate synthase (TS) encoding DNA but use different strategies to distinguish intronless from intron containing substrates. I-TevI discriminates between substrates at the level of DNA binding, as its recognition sequence is centered on the intron IS. I-BmoI, in contrast, possesses a very asymmetric recognition sequence with respect to the intron IS, binds both intron-containing and intronless TS-encoding substrates, but efficiently cleaves only intronless substrate. Here, we show that I-BmoI is extremely tolerant of multiple substitutions around its cleavage sites and has a low specific activity. However, a single G-C base pair, at position -2 of a 39 base pair recognition sequence, is a major determinant for cleavage efficiency and distinguishes intronless from intron-containing alleles. Strikingly, this G-C base pair is universally conserved in phylogenetically diverse TS-coding sequences; this finding suggests that I-BmoI has evolved exquisite cleavage requirements to maximize the potential to spread to variant intronless alleles, while minimizing cleavage at its own intron-containing allele. PMID- 12781138 TI - A Y chromosome census of the British Isles. AB - The degree of population replacement in the British Isles associated with cultural changes has been extensively debated. Recent work has demonstrated that comparisons of genetic variation in the British Isles and on the European Continent can illuminate specific demographic processes in the history of the British Isles. For example, Wilson et al. used the similarity of Basque and Celtic Y chromosomes to argue for genetic continuity from the Upper Palaeolithic to the present in the paternal history of these populations (see also ). Differences in the Y chromosome composition of these groups also suggested genetic signatures of Norwegian influence in the Orkney Islands north of the Scottish mainland, an important center of Viking activities between 800 and 1300 A.D. More recently, Weale et al. argued for substantial Anglo-Saxon male migration into central England based on the analysis of eight British sample sets collected on an east-west transect across England and Wales. To provide a more complete assessment of the paternal genetic history of the British Isles, we have compared the Y chromosome composition of multiple geographically distant British sample sets with collections from Norway (two sites), Denmark, and Germany and with collections from central Ireland, representing, respectively, the putative invading and the indigenous populations. By analyzing 1772 Y chromosomes from 25 predominantly small urban locations, we found that different parts of the British Isles have sharply different paternal histories; the degree of population replacement and genetic continuity shows systematic variation across the sampled areas. PMID- 12781139 TI - Top-down modulation of lateral interactions in early vision: does attention affect integration of the whole or just perception of the parts? AB - Attention can modulate sensitivity to local stimuli in early vision. But, can attention also modulate integration of local stimuli into global visual patterns? We recently measured effects of attention on the phenomenon of lateral interactions between collinear elements, commonly thought to reflect long-range mechanisms in early visual cortex underlying contour integration. We showed improved detection of low-contrast central Gabor targets in the context of collinear flankers, but only when the collinear flankers were attended for a secondary task rather than ignored in favor of an orthogonal flanker pair. Here, we contrast two hypotheses for how attention might modulate flanker influences on the target: by changing just local sensitivity to the flankers themselves (flanker-modulation-only hypothesis), or by weighting integrative connections between flanker and target (connection-weighting hypothesis). Modeled on the known nonlinear dependence of target visibility on collinear flanker contrast, the first hypothesis predicts that an increase in physical flanker contrast should readily offset any reduction in their effective contrast when ignored, thus eliminating attentional modulation. Conversely, the second hypothesis predicts that attentional modulation should persist even for the highest flanker contrasts. Our results showed the latter outcome and indicated that attention modulates flanker-target integration, rather than just processing of local flanker elements. PMID- 12781144 TI - Memory on the move. PMID- 12781145 TI - IP3 receptors. PMID- 12781146 TI - Calcium-induced calcium release. PMID- 12781149 TI - Dendritic arbors: a tale of living tiles. AB - What governs the shape and size of a neuron's dendritic arbor? Part of the answer lies in the rules that govern interactions between dendrites. Interesting new insights into these rules have come from two recent studies on the embryonic sensory system of Drosophila. PMID- 12781150 TI - Microtubule cytoskeleton: navigating the intracellular landscape. AB - Recent studies have significantly advanced our understanding of how a dividing cell asymmetrically positions the mitotic spindle--a key process in metazoan development--while maintaining a dynamic spindle state that can respond and reorient when necessary. PMID- 12781151 TI - Synaptic transmission: functional autapses in the cortex. AB - Autapses--synapses made by a neuron onto itself--have often been observed anatomically in vivo, but their purpose has remained uncertain. A recent paper describes functional autapses in the cerebral cortex and gives clues to their physiological role. PMID- 12781152 TI - Evolution: have wings come, gone and come again? AB - Can complex traits be re-evolved by lineages that have lost them? Phylogenetic study now suggests that wings may indeed have reappeared several times within the ancestrally wingless stick insects. PMID- 12781153 TI - Telomere replication: an Est fest. AB - The search for subunits of the telomerase enzyme has uncovered orthologs of the budding years Est1 protein in several species, including humans. Thus, positive regulation of telomerase by Est1 appears to be a widely utilized mechanism for maintaining telomere length homeostasis. PMID- 12781154 TI - Cognitive physiology: moving the mind's eye before the head's eye. AB - Under natural conditions, shifts of spatial attention are often followed by matching eye movement. Recent evidence suggests that this close coupling is reflected in the ability of the same cortical area to shift eye position and the locus of attention. PMID- 12781156 TI - Cognitive neuroscience: numerate neurons. AB - Whether the neuronal encoding of number is linear or logarithmic divides cognitive neuroscientists working on mathematical cognition. Recordings from the prefrontal cortex of the monkey support the logarithmic hypothesis. Similarities between number and the coding of other quantities are also beginning to become apparent. PMID- 12781155 TI - Mitotic entry: tipping the balance. AB - In the eukaryotic cell cycle, the G2-M transition is regulated by a counteracting kinase/phosphatase mechanism. A novel player, Tome-1, has now been found to trigger entry into mitosis by interfering with this kinase/phosphatase 'switch'. PMID- 12781157 TI - Captivating capture: how microtubules attach to kinetochores. AB - Accurate chromosome segregation is essential to ensure genomic stability because the aneuploidy that results from segregation errors leads to birth defects and contributes to the development of cancer. Chromosome segregation is directed by the kinetochore, the chromosomal site of attachment to dynamic polymers called microtubules (MTs). Although the fidelity of chromosome segregation depends on precise interactions between kinetochores and MTs, it is still unclear how this interaction is mediated and regulated. Here we discuss current progress in determining how kinetochores assemble and attach to MTs during mitosis as well as how they correct errors. PMID- 12781158 TI - Consumers' health perceptions of three types of milk: a survey in Australia. AB - A survey was conducted among 345 randomly selected shoppers in Melbourne, Australia, to identify their perceptions of the healthiness of whole milk, reduced fat milk and soy milk and to investigate demographic influences on health perceptions and types of milk consumption. The survey revealed major differences in shoppers' perceptions of the three types of milk along the themes of bone health, weight control, disease prevention, and, allergy and disease causation. Generally whole milk was perceived more negatively than soy or reduced fat milk. There were few demographic differences in the shoppers' perceptions of the three products. Reported consumption of reduced fat milk was more frequent among women and elderly people, and whole milk consumption was positively related to parenthood. The findings suggest that previous marketing and education campaigns have been effective but the widespread uncertainty and erroneous beliefs about milk products suggest that many consumers have not assimilated new findings about milk products. This represents a challenge for industry and public health practitioners. PMID- 12781159 TI - Similar effects of foods high in protein, carbohydrate and fat on subsequent spontaneous food intake in healthy individuals. AB - Pre-loads high in protein, as compared to carbohydrate and fat, produce greater satiety and reduce food intake after a fixed time interval. This study investigated the effect of macronutrient composition on spontaneous eating behaviour. On four separate occasions, 16 fasted, healthy, non-obese men, blinded to the true purpose of the study, consumed iso-energetic ( approximately 3MJ) yoghurt-based pre-loads of equivalent weight ( approximately 0.5 kg), high in fat (40%) [HF], carbohydrate (60%) [HC] or protein (29%) [HP], and no pre-load in a randomized, single-blind fashion. Subjects ate at will from a selection of food items for the remainder of the day (7 h) with the time of food requests (h) and energy content (kJ) and macronutrient distribution (%) of food eaten recorded. The three pre-loads delayed the first spontaneous request for food by 1.5-1.8 h relative to no pre-load. Total spontaneous food intake was suppressed 29% [HP], 20% [HF] and 17% [HC] by the pre-loads. Neither the amount of food eaten per spontaneous eating episode, nor the spontaneous eating frequency differed statistically following ingestion of the different pre-loads or no pre-load. In this study, in subjects who were free to choose when as well as how much they ate, a high-protein pre-load exerted similar effects on satiety as did iso energetic high-fat and high-carbohydrate pre-loads. PMID- 12781160 TI - Choice of organic foods is related to perceived consequences for human health and to environmentally friendly behaviour. AB - We designed a questionnaire concerned with attitudes and behaviour towards organic foods, environmentally friendly behaviour (EFB), and perceived consequences of organic food choice in terms of human health, the environment and animal welfare. It was mailed in 1998 to a random nation-wide sample of 2000 Swedish citizens, ages 18-65 years, and 1154 (58%) responded. Self-reported purchase of organic foods was most strongly related to perceived benefit for human health. Performance of EFBs such as refraining from car driving was also a good predictor of purchase frequency. The results indicate that egoistic motives are better predictors of the purchase of organic foods than are altruistic motives. PMID- 12781161 TI - Effects of repeated exposure and health-related information on hedonic evaluation and acceptance of a bitter beverage. AB - The influence of exposure and information on sensory evaluation and acceptance of bitter flavor was assessed. Following sensory testing in the laboratory, subjects consumed a commercially-available bittersweet beverage once daily for 7days in a setting of their choosing, then returned to the laboratory for post-exposure tests. Hedonic ratings for the beverage increased by 68%, whereas ratings for control stimuli sampled only in the laboratory did not change. Following exposure, relationships of hedonic ratings with intensity and familiarity varied according to the context in which evaluations were made: hedonic ratings were correlated with intensity in a familiar setting and with familiarity in an unfamiliar setting. Health-related information had no effect on perceptual changes that accompanied exposure, but did tend to increase a behavioral measure of acceptability, suggesting that information may have a greater effect on behavior than on hedonics. Together, the data suggest that repeated exposure can enhance hedonic evaluation of a bittersweet beverage, perhaps through a learned association of flavor with post-ingestive consequences. Context may mediate this effect and studies addressing the influence of information on bitter food acceptance should include measures of consumption and evaluate information strategies, as bitter foods may be particularly resistant to cognitively-based appeals. PMID- 12781162 TI - Agreement between weekly vs. daily assessment of appetite. AB - Hunger, satiation, and other aspects of appetite are frequently measured using end-anchored line ratings. This study investigated whether a single assessment of appetite, which covered the prior week, would yield the same results obtained by assessing appetite 3 days during the week. Participants were 51 women and 9 men (age mean 44.1, SD 9.8 years; BMI mean 38.0, SD 5.2 kg/m(2)) in a study evaluating the effectiveness of four weight control programs, three of which used sibutramine (15 mg/d). On three separate days of one week, participants rated appetite for that day. During a subsequent clinic visit, participants completed the same ratings in reference to the past week. The three daily values were averaged. All correlations between weekly and averaged daily values were strongly positive (r=0.63-0.80) and statistically significant (all p<0.001). All mean daily and weekly ratings fell within 3.3-7.2 mm of each other. Thus, good agreement was observed between the average daily assessments and one weekly assessment. The weekly assessment of appetite in the clinic appears to have several benefits, including uniformity of test administration and decreased burden on participants. PMID- 12781163 TI - Relation between appetite ratings before and after a standard meal and estimates of daily energy intake in obese and reduced obese individuals. AB - The aim of the present study was to relate appetite ratings before and after a standard breakfast to estimates of daily energy intake, before and after weight loss obese men and women. Nineteen obese subjects (9 men and 10 women) took part in a 15-week drug-based weight-loss program coupled to energy intake restriction. Body weight and body composition were significantly decreased in men and women. Both before and after the weight loss program, desire to eat, hunger, fullness and prospective food consumption (PFC) were measured after an overnight fast and at 10-min intervals in the hour following the ingestion of a standardized breakfast. Energy intakes were also measured and reported before and after weight loss. Fasting desire to eat and postprandial area under the curve (AUC) for hunger were significantly increased (p<0.05) after the intervention. No association was observed between measured or reported energy intakes and appetite ratings before weight loss in either men or women. Reported energy intake was not associated with appetite sensations after weight loss either. In contrast, measured energy intake was significantly associated with postprandial AUC for fullness (r=-0.90, p<0.01) and PFC (r=0.80, p<0.01) in men at the end of the program. In stepwise multiple regression analysis, only postprandial AUC for PFC contributed independently to the variance of measured energy intake after weight loss (r(2)=0.60, p=0.01). This study did not show consistent associations between averaged appetite ratings after a meal and daily energy intake, either before or after weight loss. PMID- 12781164 TI - Impairments in working memory associated with naturalistic dieting in women: no relationship between task performance and urinary 5-HIAA levels. AB - The present study investigated the question of whether the previously observed impairments of working memory characteristic of dieting to lose weight can be explained in terms of preoccupying cognitions relating to body shape or to alterations in serotonergic function resulting from a low dietary intake of tryptophan. The population comprised female non-dieting, lower restrained eaters (N=23), non-dieting higher restrained eaters (N=11) and current dieters (N=19). Each participant completed three tasks, each of which selectively loaded on to a different sub-component of working memory. The tasks comprised the Tower of London task, a letter string recall task and a mental rotation task. In addition, all participants completed self-report measures of body shape concern and affective state. Serotonin turnover was assessed by means of 24 h urine sample collection for each participant on their day of testing. This was analysed (via HPLC) for levels of the main serotonin metabolite 5-HIAA.The results of the present study broadly replicated previous findings of a Central Executive and Phonological Loop (but not Visuo-Spatial Sketchpad) deficit in those subjects who reported themselves to be currently dieting. Tower of London task performance also significantly correlated with self-reported feelings of fatness and body shape disparagement. There were no group differences in 5-HIAA levels nor did 5 HIAA levels correlate with task performance. However, there was a significant negative correlation between 5-HIAA levels and self-reported depression. These results support the hypothesis that the variables mediating this deficit are preoccupying cognitions concerning body shape. They do not support the hypothesis that the serotonergic function of dieters is compromised, although this conclusion is tentative. PMID- 12781165 TI - Increasing children's acceptance of vegetables; a randomized trial of parent-led exposure. AB - Despite considerable epidemiological evidence of the health benefits of a diet high in fruit and vegetables, consumption in pre-school children remains well below recommended levels. This study evaluated the effectiveness of an exposure based intervention, carried out by parents in the home, in increasing children's liking for a previously disliked vegetable. 156 parents of 2-6 year old children were randomly assigned to Exposure, Information or Control groups after a pre intervention taste test at which a 'target' vegetable was selected. Parents in the Exposure group gave their child a taste of this vegetable daily for 14 days, parents in the Information group were given nutritional advice and a leaflet, and parents in the Control group received no further intervention. All participants took part in a post-intervention taste test. Greater increases in liking, ranking and consumption of the 'target' vegetable from pre- to post-intervention occurred in the Exposure group than in either of the other two groups. Only the Exposure group showed significant increases across all three outcomes. It can be concluded that a parent-led, exposure-based intervention involving daily tasting of a vegetable holds promise for improving children's acceptance of vegetables. These findings suggest a parental advice strategy which could be disseminated directly to parents or by health professionals. PMID- 12781167 TI - Food intake and circadian rhythms in shift workers with a high workload. AB - Shift work is associated with nutritional and health problems. In the present study, the food intake of garbage collectors of the city of Florianopolis (Brazil) was investigated using a dietary survey method based on meal recording during 24 h and adapted for the Brazilian food context. Three different shifts (morning, afternoon, and night) were compared (n=22 per shift). Age, body weight and body mass index (BMI) were similar for all groups. Daily energy expenditure was high in all three shifts, especially in morning shift workers. No difference in daily energy intake was found, in spite of differences in food choices and circadian ingestion rhythms. Energy intake was high and close to levels previously reported in athletes. Several factors not associated with shifts had significant impact on ingestion: hour of the day, time since the last meal, age, and BMI. Ingested foods were analyzed in groups based on nutrient content. Shifts significantly influenced intake of starches, alcoholic drinks, and sweets. In different periods of the day, food and nutrient intake were considerably affected by shifts. The analysis of circadian distribution of food choices and nutrient intake is important in shift workers, because total daily intake may not reveal shift-associated differences. PMID- 12781166 TI - Validation and cross-national comparison of the food neophobia scale (FNS) using confirmatory factor analysis. AB - The food neophobia scale [FNS; Appetite 19 (1992) 105] has been used to assess willingness to try new foods in studies conducted around the world. Although it is tempting to compare FNS scores across these studies, appropriate psychometric analyses are required to validate the scale and allow cross-cultural comparisons. These analyses were pursued in the current study using confirmatory factor analysis in conjunction with a data analysis strategy described by Steenkamp and Baumgartner [J. Consumer Res. 25 (1998) 78] and random, representative samples drawn from the United States, Sweden and Finland. A unidimensional scale was constructed using eight of the original 10 items from the FNS, and this model provided an excellent fit to the data from the US and Swedish samples. An acceptable fit was achieved for six items when data from the US, Sweden and Finland were used. Based on these analyses, we recommend that two items from the original FNS be dropped (items 5 and 9). Elimination of additional items may be premature given the potential contributions of difference in sampling and testing methodology associated with data collection from the three samples. Future efforts to develop a FNS for cross-national comparisons should target the development of a scale with 14-16 items so that dropping several items from a model (due to translation or other problems) allows retention of a sufficient number of items to insure a robust test. However, even with only six items, our results supported the conclusion that people from Sweden are generally more willing to try novel foods as compared to people from the US and Finland. Future studies should focus on the source of this enhanced willingness to try novel foods among the Swedes and the potential use of this information in the development of programs aimed at facilitating dietary change. PMID- 12781168 TI - Cognitive dietary restraint is associated with eating behaviors, lifestyle practices, personality characteristics and menstrual irregularity in college women. AB - This study characterized associations of restraint with selected physical, lifestyle, personality and menstrual cycle characteristics in female university students. The survey instrument, distributed to 1350 women, included standardized questionnaires (Three-Factor Eating Questionnaire, Perceived Stress Scale and Rosenberg's Self-esteem Scale), and assessed weight and dieting history, exercise, lifestyle characteristics, menstrual cycle characteristics and whether participants were following vegetarian diets. Among the 596 respondents included in the analysis (44%), women with high (n=145), medium (n=262) or low (n=189) restraint had similar ages, heights and weights. Despite this, compared to women with low scores, those with high scores exercised more (4.6+/-5.3 vs. 3.2+/-3.5 h/wk), were more likely to be vegetarian (14.5 vs. 3.7%), have a history of eating disorders (13.7 vs. 1.2%), be currently trying to lose weight (80.3 vs. 15.3%), report irregular menstrual cycles (34.7 vs. 17.0%), and have scores reflecting lower self-esteem and higher perceived stress. Menstrual irregularity was an independent predictor of restraint score, and restraint score was the only variable to differentiate women with regular and irregular menstrual cycles. We conclude that women with high restraint may use a combination of behavioral strategies for weight control, and differ from women with low restraint scores in personality characteristics and weight history. Some of these behaviors or characteristics may influence menstrual function. PMID- 12781169 TI - Food craving patterns in Egypt: comparisons with North America and Spain. PMID- 12781172 TI - Synthesis, stereochemical assignment and biological activity of a novel series of C-4" modified aza-macrolides. AB - Modification of the cladinose C-4" position via manipulation of the corresponding keto derivatives afforded two stereochemically pure series of compounds. The synthesis and structure determination of these compounds is described within. The in vitro and in vivo biological activity of this novel series of C-4" modified macrolides is also described. PMID- 12781173 TI - Identification of a dual histamine H1/H3 receptor ligand based on the H1 antagonist chlorpheniramine. AB - Combining the first generation H(1) antihistamine chlorpheniramine (1) with H(3) ligands of the alkylamine type has led to the identification of compound 9d, a dual ligand of both the H(1) and H(3) receptors. PMID- 12781174 TI - Sphingomyelin analogues as inhibitors of sphingomyelinase. AB - To search for neutral sphingomyelinase inhibitors we designed and synthesized hydrolytically stable analogues of sphingomyelin. The novel compounds 8 and 9 which were replaced the phosphodiester moiety of sphingomyelin with the carbamate moiety showed inhibitory activity with an IC(50) value of micro M on neutral sphingomyelinase in rat brain microsomes. Compound 8i showed a selective neutral sphingomyelinase inhibitory activity. PMID- 12781175 TI - Chemoenzymatic synthesis of (E)-3,7-dimethyl-2-octene-1,8-diol isolated from the hairpencils of male Danaus chrysippus (African Monarch). AB - The synthesis of (E)-3,7-dimethyl-2-octene-1,8-diol (1), which was isolated from the hairpencils of male Danaus chrysippus (African Monarch), was investigated. The key step of the sequence involves asymmetric desymmetrization of the 1,3 propanediol 7 with lipase, in which high enantioselectivity was observed. Total synthesis afforded (S)-1 in 12 steps and 26% overall yield from readily available geraniol. PMID- 12781176 TI - Novel water soluble 2,6-dimethoxyphenyl ester derivatives with intravenous anaesthetic activity. AB - A number of water soluble bis-amino-2,6-dimethoxyphenyl ester derivatives were found to exhibit improved anaesthetic activity in mice relative to propofol 1. Of the analogues disclosed, 44 was further profiled in rodents and found to be a superior agent to propofol for the induction and maintenance of anaesthesia. PMID- 12781177 TI - Hemisynthesis and preliminary evaluation of novel endocannabinoid analogues. AB - Three new endocannabinoid analogues in which amide moiety was replaced either by oxomethylene group or ester moiety with simultaneous substitution of both alpha hydrogens with methyl groups were synthesized and their abilities to interact with CB1-receptor and FAAH were investigated. PMID- 12781178 TI - Dihydroquinolines with amine-containing side chains as potent n-NOS inhibitors. AB - Dihydroquinolines with aminoalkyl side chains have been synthesized and have been shown to be potent n-NOS inhibitors. A marked selectivity versus e-NOS of up to approximately 300-fold was observed, whereas i-NOS was moderately inhibited. PMID- 12781179 TI - Unexpected inhibition of S-adenosyl-L-homocysteine hydrolase by a guanosine nucleoside. AB - A series of shape-modified flexible nucleosides ('fleximers', 1, 2, and 3) was modeled, synthesized and subsequently assayed against S-adenosyl-L-homocysteine hydrolase (SAHase). No inhibitory activity was observed for the adenosine fleximer, which served as a substrate, but moderate inhibitory activity was exhibited by the guanosine fleximers. This is the first known report of a guanosine nucleoside analogue possessing activity against SAHase. PMID- 12781180 TI - Structure-activity relationships in a series of NPY Y5 antagonists: 3-amido-9 ethylcarbazoles, core-modified analogues and amide isosteres. AB - Beginning with carbazole 1a, the amide and alkyl substituents were optimized to maintain potency while adding solubilizing groups. Efforts to replace the 3-amino 9-ethylcarbazole core, a known carcinogen, used the SAR generated in the carbazole series for guidance and led to the synthesis of a number of core modified analogues. In addition, an isosteric series, in which the amide was replaced with an imidazole, was prepared. Two potent new series lacking the putative toxicophore were identified from these endeavors. PMID- 12781181 TI - The role of 2',3'-unsaturation on the antiviral activity of anti-HIV nucleosides against 3TC-resistant mutant (M184V). AB - Molecular modeling studies show that the 2',3'-double bond of the sugar moiety of various 2',3'-unsaturated nucleosides interacts with the aromatic moiety of Tyr115 of HIV-1 reverse transcriptase (RT) by hydrophobic pi-pi interaction. In 3TC-resistant mutant (M184V) RT, 2'-fluoro-2',3'-unsaturated nucleosides with a bulky 4'-substituent experience significant steric hindrance with the side chain of Val184. PMID- 12781182 TI - Arylaminoethyl amides as inhibitors of the cysteine protease cathepsin K investigating P1' substituents. AB - Modeling, synthesis and in vitro activities of a series of arylaminoethyl amide based inhibitors of the cysteine protease cathepsin K are described. PMID- 12781183 TI - The discovery of a new class of large-conductance Ca2+-activated K+ channel opener targeted for overactive bladder: synthesis and structure-activity relationships of 2-amino-4-azaindoles. AB - 2-Amino-4-azaindoles have been identified as a structurally novel class of BK(Ca) channel openers. Their synthesis from 2-chloro-3-nitropyridine is described together with their in vitro properties assessed by 86Rb(+) efflux and whole-cell patch-clamp assays using HEK293 cells stably transfected with the BK(Ca) alpha subunit. In vitro functional characterization of BK(Ca) channel opening activity was also assessed by measurement of relaxation of smooth muscle tissue strips obtained from Landrace pig bladders. The preliminary SAR data indicate the importance of steric bulk around the 2-amino substituent. PMID- 12781184 TI - A study on the synthesis of antiangiogenic (+)-coronarin A and congeners from (+) sclareolide. AB - Coronarin A 1, epi-coronarin A 2 and some synthetic intermediates 14a and 14b synthesized from sclareolide exhibit good growth inhibition activities on HUVEC proliferation. In particular, coronarin A 1 and epi-coronarin A 2 effectively suppressed the growth factor induced tube formation of HUVEC at the concentration of 10 micro g/mL. PMID- 12781185 TI - 1,4-Dihydroxy-2,3-dioxatricyclo[8.4.0.0(4,9)]tetradecane and derivatives with in vitro activity against Plasmodium falciparum, Trypanasoma b brucei, Trypanasoma cruzi, and Leishmaniasis infantum. AB - 1,4-Dihydroxy-2,3-dioxatricyclo[8.4.0.0(4,9)]tetradecane and derivatives have been synthesised and their in vitro activity against Plasmodium falciparum (malaria) Ghana, Trypanasoma b brucei (sleeping sickness) TB-1, and Trypanasoma cruzi (Chagas' disease) TC-1, and Leishmaniasis infantum (leishmaniasis) L1 parasite strains has been assessed. PMID- 12781186 TI - N-ferrocenylmethyl, N'-methyl-2-substituted benzimidazolium iodide salts with in vitro activity against the Leishmania infantum parasite strain L1. AB - Herein, we disclose results of our research into a class of benzimidazolium compounds active against the Leishmania infantum parasite strain L1. We have discovered that N-ferrocenylmethyl, N'-methyl-2-aryl (or styryl) benzimidazolium iodide salts show in vitro activity against this strain. PMID- 12781187 TI - Discovery of selective phosphonamide-based inhibitors of tumor necrosis factor alpha converting enzyme (TACE). AB - A novel series of phosphonamide-based inhibitors of tumor necrosis factor-alpha converting enzyme (TACE) was discovered by structural modification of tetrahydroisoquinoline derivative 1b, which was extremely weak inhibitor of TACE. (S)-Isomer at the phosphorus atom (7b) displayed potent inhibition for TACE, while selectivity sparing MMP-1, -3, and -9. PMID- 12781188 TI - Chemical synthesis and cytotoxicity of dihydroxylated cyclopentenone analogues of neocarzinostatin chromophore. AB - Compounds containing the naphthoate moiety of Neocarzinostatin chromophore or 2 hydroxynaphthoate have been synthesized and evaluated for cytotoxic activity against a leukemia cell line and a small panel of human-tumor cell lines. Those compounds containing a cyclopentenone moiety were active, with the carbonyl group being essential for biological activity. PMID- 12781189 TI - D-Phe-Pro-Arg type thrombin inhibitors: unexpected selectivity by modification of the P1 moiety. AB - Synthesis of thrombin inhibitors and their binding mode to thrombin is described. Modification of the P1 moiety leads to an increased selectivity versus trypsin. The observed selectivity is discussed in view of their thrombin-inhibitor complex X-ray structures. PMID- 12781190 TI - Discovery of N-hydroxy-2-(2-oxo-3-pyrrolidinyl)acetamides as potent and selective inhibitors of tumor necrosis factor-alpha converting enzyme (TACE). AB - New inhibitors of tumor necrosis factor-alpha converting enzyme (TACE) were discovered using an N-hydroxy-2-(2-oxo-3-pyrrolidinyl)acetamide scaffold. The series was found to be potent in a porcine TACE (pTACE) assay with IC(50)s typically below 5 nM. For most compounds, selectivity for pTACE relative to MMP 1,-2, and -9 is at least 300-fold. Compound 2o was potent in inhibition of TNFalpha production in a human whole blood assay (WBA) with an IC(50) of 0.42 micro M. PMID- 12781191 TI - Inhibition of mandelate racemase by alpha-fluorobenzylphosphonates. AB - Mandelate racemase catalyzes the interconversion of the enantiomers of mandelic acid. The enzyme binds the intermediate analogues (R)- and (S)-alpha fluorobenzylphosphonate, and alpha,alpha-difluorobenzylphosphonate with 100-2500 times less affinity than it exhibits for (R,S)-alpha-hydroxybenzylphosphonate at pH 7.5. This apparent low affinity, relative to that of alpha hydroxybenzylphosphonate, arises from the altered pKa values of the alpha fluorobenzylphosphonates. For example, (S)-alpha-fluorobenzylphosphonate is bound with the same affinity as the substrate at pH 7.5, but this affinity is increased approximately 6-fold at pH 6.3. PMID- 12781192 TI - Synthesis of 2-deoxy-2-C-alkylglucosides of myo-inositol as possible inhibitors of a N-deacetylase enzyme in the biosynthesis of mycothiol. AB - Two new analogues of 1-D-1-O-(2-acetamido-2-deoxy-alpha-D-glucopyranosyl)-myo inositol, a biosynthetic intermediate in the production of mycothiol in the Mycobacteria have been synthesized. Both the 2-deoxy-2-C-(2'-hydroxypropyl)-D glucoside 5, and the 2-deoxy-2-C-(2'-oxopropyl)-D-glucoside 6, are derived from fully benzylated 1-D-1-O-(2-C-allyl-2-deoxy)-D-glucopyranosyl)-myo-inositol 20, readily assembled via a protected 2-C-allyl-2-deoxyglucosyl fluoride. Both 5 and 6 inhibit the incorporation of [3H]inositol by whole cells of Mycobacterium smegmatis into a number of metabolites which contain inositol. PMID- 12781193 TI - 3,4-disubstituted azetidinones as selective inhibitors of the cysteine protease cathepsin K. Exploring P2 elements for selectivity. AB - A novel series of 3,4-disubstituted azetidinones based inhibitors of the cysteine protease cathepsin K (Cat K) has been identified. Although not optimized, some of these compounds show at least 100-fold selectivity against other cathepsins. The use of cyclic moieties as P2 elements has proven to be crucial to achieve a high degree of selectivity. PMID- 12781194 TI - 3-(2-carboxyethyl)-4,6-dichloro-1H-indole-2-carboxylic acid: an allosteric inhibitor of fructose-1,6-bisphosphatase at the AMP site. AB - 3-(2-Carboxyethyl)-4,6-dichloro-1H-indole-2-carboxylic acid (MDL-29951), an antagonist of the glycine site of the NMDA receptor, has been found to be an allosteric inhibitor of the enzyme fructose 1,6-bisphosphatase. The compound binds at the AMP regulatory site by X-ray crystallography. This represents a new approach to inhibition of fructose 1,6-bisphosphatase and serves as a lead for further drug design. PMID- 12781195 TI - Inhibitors of inosine monophosphate dehydrogenase: SARs about the N-[3-Methoxy-4 (5-oxazolyl)phenyl moiety. AB - The first reported structure-activity relationships (SARs) about the N-[3-methoxy 4-(5-oxazolyl)phenyl moiety for a series of recently disclosed inosine monophosphate dehydrogenase (IMPDH) inhibitors are described. The syntheses and in vitro inhibitory values for IMPDH II, and T-cell proliferation (for select analogues) are given. PMID- 12781196 TI - The activity of diguanidino and 'reversed' diamidino 2,5-diarylfurans versus Trypanosoma cruzi and Leishmania donovani. AB - The in vitro activity of 20 dicationic molecules containing either diguanidino or reversed amidine cationic groups were evaluated versus Trypanosoma cruzi and Leishmania donovani. The most active compounds were in the reversed amidine series and six exhibited IC(50) values of less than 1 micro mol versus T. cruzi and five gave similar values versus L. donovani. PMID- 12781197 TI - Synthesis and biological activity of 5-methylidene 1,2-dihydrochromeno[3,4 f]quinoline derivatives as progesterone receptor modulators. AB - A series of 5-methylidene 1,2-dihydrochromeno[3,4-f]quinoline derivatives were synthesized and tested in biological assays to evaluate scope and limitations of the nonsteroidal SPRM pharmacophore (3). A number of orally available highly potent nonsteroidal modulators were identified by modification of the substituents at 5-methylidene position. PMID- 12781198 TI - Development of progesterone receptor antagonists from 1,2-dihydrochromeno[3,4 f]quinoline agonist pharmacophore. AB - A series of 1,2-dihydrochromeno[3,4-f]quinoline derivatives was synthesized and tested in biological assays to evaluate the nonsteroidal progesterone receptor modulator pharmacophore (4) as antiprogestins. A number of potent analogues were identified by modification of the substituents at the D-ring. PMID- 12781199 TI - Urea small molecule agonists on mouse melanocortin receptors. AB - A series of urea compounds based on the tripeptide Phe-Trp-Lys were synthesized and pharmacologically characterized at the mouse melanocortin receptors. The results include identification of novel melanocortin receptor agonists with potencies ranging from nanomolar to micromolar. PMID- 12781200 TI - Design and synthesis of phosphotyrosine mimetics. AB - Selective inhibitors of protein tyrosine phosphatases (PTPases) are of great interest as therapeutic agents and research tools. Several phenylalanine derivatives (1, 2) designed as phosphotyrosine mimetics or irreversible active site inhibitors were successfully synthesized, then incorporated into a combinatorial library based on a peptidomimetic beta-strand template. PMID- 12781202 TI - Inhibition of various glutathione S-transferase isoenzymes by RRR-alpha tocopherol. AB - The activity of human cytosolic glutathione S-transferases (GSTs) can positively or negatively be changed by various compounds. It is for instance known that RRR alpha-tocopherol inhibits GST P1-1 [Haaften van R.I.M. et al. (2001) Alpha tocopherol inhibits human glutathione S-transferase pi. BBRC 280, 631-633]. The effect of RRR-alpha-tocopherol on the other isoenzymes of GST in purified forms of the isoenzymes and in human liver cytosol (GST M and GST A) and lysate of human erythrocytes (GST P) is studied. It is found that all isoenzymes (purified enzymes and enzymes present in homogenates) are inhibited, in a concentration dependent way, by RRR-alpha-tocopherol. GST P is in both cases inhibited with the highest potency compared to the other isoenzymes. It also appeared that the purified GST P1-1 isoenzyme is non-competitively inhibited by RRR-alpha tocopherol. The IC(50) values of RRR-alpha-tocopherol for the purified isoenzymes of GST are much lower compared to the IC(50) values for human lysate and human liver cytosol. This is probably due to binding of RRR-alpha-tocopherol to proteins, e.g. albumin and hemoglobin, with higher affinity than to GST; so more RRR-alpha-tocopherol is needed to inhibit the enzyme. However, the inhibition of GSTs by RRR-alpha-tocopherol can still be of physiological relevance, because due to dermal application of cosmetic products very high concentrations vitamin E can be reached in the skin, where GST P1-1 is present. RRR-alpha-tocopherol might also be a good lead compound for the development of a new class of inhibitors of GST that can be used as adjuvant in cancer therapy. PMID- 12781203 TI - Flow cytometric assessment of changes in rat sperm mitochondrial function after treatment with pentachlorophenol. AB - The fluorophore 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolyl carbocyanine iodide (JC-1) localizes to the mitochondria and is affected by membrane potential, fluorescing bright orange when the membrane potential is high and green when mitochondrial membrane potential is low. The present study used flow cytometric analysis of JC-1 staining patterns of large numbers of spermatozoa to detect chemical-induced alterations of sperm mitochondrial membrane potential. Cauda epididymal rat spermatozoa were incubated with pentachlorophenol (PCP; 0.1 microM or 1.0 microM), a known uncoupler of mitochondrial oxidative phosphorylation. Microscopic evaluation showed that the midpiece (mitochondrial location) of live, highly motile spermatozoa stained bright orange, while the midpiece of live, non-motile spermatozoa stained green. The midpiece of slightly or non-progressively motile spermatozoa stained a faint orange-green. The percentage of spermatozoa stained bright orange and the total percentage of spermatozoa stained orange (bright orange+faint orange) in the control samples of spermatozoa were significantly higher (P<0.001) than in the 0.1 microM and 1.0 microM PCP treated samples. These data indicate that sperm mitochondrial membrane potential is highly sensitive to the uncoupling effects of PCP and that JC-1 staining and flow cytometric analysis may be a sensitive assay to detect the effect of toxicants on rat sperm mitochondrial function. PMID- 12781204 TI - Effects of 3,3',4,4',5-pentachlorobiphenyl, a coplanar polychlorinated biphenyl congener, on cultured neonatal mouse testis. AB - 3,3',4,4',5-Pentachlorobiphenyl (PCB126), a congener with a planar configuration, has been established to have relatively strong toxicities similar to those of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) via aryl hydrocarbon receptors. We investigated the effects of this coplanar PCB on mammalian early spermatogenesis and steroidogenesis in a mouse neonatal testicular organ culture system. Testes collected from newborn mice were subjected to organ culture in medium containing 0, 10, 100 or 1000 nM PCB126. Histochemical analysis revealed that the BrdU labeling indices of both spermatogenic cells and Sertoli cells were unchanged in all testis specimens exposed to the coplanar PCB. CYP1A1 and steroidogenic enzymes (P450scc, P450c17, 3beta-HSD and 17beta-HSD) mRNA levels were determined by semiquantitative RT-PCR. The CYP1A1 mRNA level in cultured testis was significantly increased by PCB126 in a dose-dependent manner. Although mRNA levels of 3beta-HSD and 17beta-HSD were unchanged, the P450scc mRNA level was significantly down-regulated by PCB126 in a dose-dependent manner. In contrast, the P450c17 mRNA level was significantly higher in 1000 nM PCB126-exposed testis than in control testis. These results suggest that the coplanar PCB does not alter the proliferative activity of spermatogenic cells and Sertoli cells in neonatal testis, but that it directly affects the expression of steroidogenic enzyme genes. PMID- 12781205 TI - Myeloid clonogenic assays for comparison of the in vitro toxicity of alkylating agents. AB - A battery of clonal assays for myeloid progenitor cells (HPP-CFC, CFU-gemm, CFU gm, CFU-g) was utilized to evaluate the myelotoxicity of a series of alkylating agents representing the spectrum of clinical times to nadir. Bone marrow aspirates from normal volunteers were incubated with mechlorethamine, busulfan, melphalan, carmustine or lomustine for 1 h and then cultured in methylcellulose with 30% serum and cytokines. There was a concentration-dependent inhibition of colony formation and often a differential toxicity to the myeloid progenitors with the alkylators tested. On a molar basis, mechlorethamine and melphalan were the most toxic of the alkylator drugs to the myeloid precursors. The most sensitive progenitor was CFU-gemm with the lowest inhibitory concentration IC(70) concentrations for mechlorethamine, melphalan, carmustine and lomustine. Generally, there was great similarity for drug effects between CFU-g and CFU-gm with overlapping inhibition curves. HPP-CFC proved to be the least sensitive of the progenitors to the toxic actions of the drugs. While there was no correlation between the time to clinical neutropenic nadir and the most sensitive progenitor in the clonal assays, the CFU-gm assay remains a suitable method for determining the myelotoxic potential of cytotoxic agents. PMID- 12781206 TI - Effect of 5-methoxypsoralen (5-MOP) on cell apoptosis and cell cycle in human hepatocellular carcinoma cell line. AB - The chemopreventive role of 5-methoxypsoralen (5-MOP) in the human hepatocellular carcinoma (HCC) cell line was investigated by studying the regulation of proliferation and apoptosis in HCC (J5) cells. Morphological analysis, cell viability assay, DNA analysis and cell-cycle analysis suggest that there are at least three modes of the suppressive effects shown by 5-MOP: (a) kills J5 cells directly; (b) induces apoptosis by arresting J5 cells at the G2/M phase in the cell cycle; (c) induces apoptosis through an independent pathway with cell-cycle arrest at 24-72 h of exposure. Further immunoblot analysis demonstrated that inhibition of cyclin B1 by 5-MOP may play an important role in G2/M arrest of J5 cells and provides an additional way to prevent cells from entering the M phase and undergoing apoptosis. 5-MOP therefore appears to exert its anticarcinogenic properties by cytotoxic effect, inducing apoptosis and inhibiting proliferation in the human hepatocellular carcinoma cell line. PMID- 12781207 TI - Percutaneous absorption of 2,6-di-tert-butyl-4-nitrophenol (DBNP) in isolated perfused porcine skin. AB - DBNP (2,6-di-tert-butyl-4-nitrophenol) has been reported as a potential contaminant in submarines. This yellow substance forms when lubrication oil mist containing the antioxidant additive 2,6-di-tert-butylphenol passes through an electrostatic precipitator and is nitrated. Percutaneous absorption of 14C-DBNP was assessed in the isolated perfused porcine skin flap (IPPSF). Four treatments were studied (n=4 flaps/treatment): 40.0 microgram/cm(2) in 100% ethanol; 40.0 microgram/cm(2) in 85% ethanol/15% H(2)O; 4.0 microgram/cm(2) in 100% ethanol; and 4.0 microgram/cm(2) in 85% ethanol/15% water. DBNP absorption was minimal across all treatment groups, with the highest absorption detected being only 1.08% applied dose in an aqueous ethanol group. The highest mass of 14C-DBNP absorbed was only 0.5 microgram. The majority of the applied dose remained on the surface of the skin. This suggests that there is minimal dermal exposure of DBNP when exposed topically to skin. PMID- 12781208 TI - Morphological characterisation of BGM (Buffalo Green Monkey) cell line exposed to low doses of cadmium chloride. AB - Morphological changes in the Buffalo Green Monkey (BGM) cell line after exposure to a subcytotoxic dose (0.062 mM, equivalent to EC(10)-effective concentration 10%) of cadmium chloride have been evaluated. Cells were exposed for 24 h and the effects observed at the ultrastructural level by transmission and scanning microscopy. Using transmission electron microscopy, the most notable findings in treated cells were the presence of intranuclear inclusion bodies and thin intracytoplasmic granules associated to myelin figures and the presence of apoptotic bodies. Other morphological alterations included cell vacuolisation and a reduced cytoplasm volume, condensation of the mitochondria and a decreased number of cytoplasmic organelles, except lysosomes and autophagic vacuoles, which increased in number. Scanning electron microscopy pointed to a cell with a disrupted perinuclear region and a decrease in the number of surface microvilli. We conclude that the BGM cell line may be considered an useful tool for toxicological studies involving cadmium. PMID- 12781209 TI - In vitro cytotoxic and anti-inflammatory effects of myrrh oil on human gingival fibroblasts and epithelial cells. AB - Limited scientific studies suggest that myrrh (Commiphora molmol) has antibacterial and anti-inflammatory activities. This study determined myrrh oil (MO) cytotoxicity to human gingival fibroblasts and epithelial cells and its effect, measured by ELISA, on interleukin (IL)-1beta-stimulated IL-6 and IL-8 production. Cell viability and cytotoxicity were determined by metabolic reduction of a tetrazolium salt to a formazan dye (MTT assay) and by release of lactate dehydrogenase (LDH) from membrane damaged (LDH release assay) cells, respectively. Based on the MTT assay, 24- and 48-h exposures to /=0.005%, maximally decreased viability of all cell lines. In the LDH release assay, exposure to /=0.0025% MO caused maximal cytotoxicity; /=0.0025% MO, probably reflective of loss of viability. At subtoxic MO levels (0.00001-0.001%), there was a significant reduction of IL-1beta-stimulated IL-6 and IL-8 production by fibroblasts, but not by epithelial cells. PMID- 12781210 TI - Analysis of interleukin-1alpha (IL-1alpha) and interleukin-8 (IL-8) expression and release in in vitro reconstructed human epidermis for the prediction of in vivo skin irritation and/or sensitization. AB - In the present study, reconstructed human epidermis (RHE) was used as an in vitro model to discriminate 1-chloro-2,4-dinitrobenzene (DNCB), nickel sulfate (NiSO(4)), oxazolone (OXA), 2,4-dinitrofluorobenzene (DNFB) and 2,4,6 trinitrobenzenesulfonic acid (TNBS) as skin sensitizers from benzalkonium chloride (BC), benzoic acid (BA) and sodium lauryl sulfate (SLS) as skin irritants. Our criteria were (a) the differential IL-1alpha and IL-8 synthesis and release (b) cytotoxicity assessment by MTT assay. When the RHE are topically treated with the sensitizers, very low levels of extra- and intracellular IL 1alpha are observed although they induce significant cytotoxicity. In contrast, they exhibit a sharp maximum of IL-8 release. In the presence of the tested irritants, we observe the inverse cytokine release profile, although they induce dose-dependent cytotoxicity profiles similar to those observed with the sensitizers. Finally, IL-1alpha mRNA upregulation is observed after topical application of both sensitizers and irritants, but only the latter significantly increase extracellular IL-1alpha. In conclusion, our results suggest that the associated determination of IL-8, with IL-1alpha, and MTT conversion are at least necessary to discriminate and classify, in a single assay, irritant and sensitizing agents and represent a potential in vitro alternative to two classical in vivo assays. PMID- 12781211 TI - Curcumin inhibited the arylamines N-acetyltransferase activity, gene expression and DNA adduct formation in human lung cancer cells (A549). AB - It is well known that N-acetyltransferase (NAT) plays an important role in the arylamine metabolism. We analysed the response of A549 human lung cancer cells for N-acetylation of 2-aminofluorene (AF) to curcumin. After curcumin treatment, the NAT activity was examined by HPLC, AF-DNA adduct formation was examined by HPLC, and NAT gene expression by polymerase chain reaction were detected. The NAT activity in the human A549 cells and cytosols was suppressed by curcumin in a dose-dependent manner. The results also demonstrated that gene expression (NAT1 mRNA) in human lung A549 tumor cells was inhibited and decreased by curcumin. After the incubation of human lung A549 tumor cells with AF with or without curcumin co-treatment, the cells were recovered and DNA was prepared and hydrolyzed to nucleotides. The adducted nucleotides were extracted into butanol and analyzation of AF-DNA adducts was done by HPLC. The results also demonstrated that curcumin decreases AF-DNA adduct formation in the human lung A549 tumor cells. PMID- 12781212 TI - Acute solvent exposure induced activation of cytochrome P4502E1 causes proximal tubular cell necrosis by oxidative stress. AB - Deliberate exposure to solvents has been associated with kidney disorders. However, the mechanism by which solvents induce renal damage after acute exposure has not been studied. Proximal tubular cell (LLC-PK1) cytotoxicity after exposure for 48 h to either 5 mM of p-xylene (XY) or toluene (TL) was compared to control (C) by cell viability (MTS assay), LDH release, DNA fragmentation, and malondialdehyde (MDA) release. CYP2E1 activity with or without a free radical scavenger (catalase-CT), or the CYP2E1 inhibitor disulfiram (DSF), was examined. Both p-xylene and toluene significantly reduced cell viability (XY 53.9 8+/-1.6 vs TL 54.8+/-0.9 vs C 102.7+/-2.1), increased CYP2E1 activity (mM/mg protein/min) (XY 3.6+/-0.5 vs TL 3.7+/-0.7 vs C 1.3+/-0.4) and MDA release (microM/mg protein) (XY 29.1+/-3.9 vs TL 12.3+/-1.4 vs C 2.8+/-0.3). LDH was increased (XY 59.9+/-3.0 vs TL 27.6+/-0.5 vs C 8.4+/-1.2), but there was no significant change in DNA fragmentation (OD/mg protein) suggesting necrosis as the predominant mode of cell death. DSF significantly attenuated CYP2E1 activity (XY+DSF 1.4+/-0.9, TL+DSF 2.3+/-0.1), LDH release (XY+DSF 45.1+/-2.0, TL+DSF 13.0+/-0.2) and MDA release (XY+DSF 4.3+/-0.5, TL+CT 6.2+/-1.1). Moreover, CT attenuated LDH release (XY+CT 36.4+/-5.1, TL+DSF 15.6+/-0.5) and MDA release (XY+DSF 5.4+/-0.7, TL+DSF 6.6+/ 1.3) in XY and TL treated cells. This study confirms the pivotal role of CYP2E1 in solvent-induced oxidative stress and necrosis in proximal tubular cells after exposure to solvent at 5 mM for 48 h. PMID- 12781213 TI - Involvement of apoptosis in hydrazine induced toxicity in rat primary hepatocytes. AB - The current study was undertaken to investigate the role of apoptosis in hydrazine induced hepatotoxicity. Hepatocytes were exposed to hydrazinium nitrate (HzN) at two doses (50 and 75 mM) for 2 h then placed in fresh HzN-free media and cultured for an additional 24 h. Post-exposure, cell viability was evaluated at several time points by lactate dehydrogenase (LDH) leakage and 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction. Markers of apoptosis (mitochondrial membrane potential, annexin binding, DNA fragmentation, caspase activation, and cytochrome c release) were measured 24 h post-exposure. The viability data showed time dependent increase in LDH leakage at 75 mM of HzN, with only a slight increase at 50 mM. MTT reduction showed a decrease in mitochondrial activity at both doses immediately after the 2 h continuous exposure. However, MTT reduction returned to normal at 50 mM while at 75 mM, MTT reduction initially recovered but then deteriorated to approximately 50% of controls at 24 h post-exposure. Based on viability data, exposure to 50 mM HzN for 2 h is a marginally toxic dose while 75 mM is a significantly toxic dose. The results for apoptosis biomarkers showed a reduction in mitochondrial membrane potential, an increase in annexin binding, an increase in total caspase activity, moderate activation of caspase-3, and release of cytochrome c. However, the appearance of DNA fragmentation in HzN exposed cells was very low compared to positive controls (cadmium and cyclosporine). The possibility that HzN induces apoptosis without the involvement of DNA fragmentation can not be ruled out. The present results, overall, suggest that apoptosis may be a contributing factor in acute HzN toxicity. PMID- 12781214 TI - Morphological and physiological changes in Tetrahymena pyriformis for the in vitro cytotoxicity assessment of Triton X-100. AB - Non-ionic surfactants such as Triton X-100 have been widely used in industrial processing and in cleaning products for almost 50 years, being effective and economic emulsifying, wetting agents, dispersants and solubilizers. Cleaning products containing these surfactants are disposed of mainly by discharge into wastewater, which receives biological treatment in wastewater treatment systems. However, surface-active agents interact with eukaryotic cell membranes leading to biological damage at high concentrations. Tetrahymena pyriformis was used here as model organism to assess the effects of Triton X-100 through a series of in vitro cytotoxicity tests. Growth rates and morphological changes were, by their simplicity and reproducibility, the simplest toxicological assays. Cytoskeleton analysis seemed to be related with phagocytosis rate. Viability was evaluated by two different tests. Calcein AM/EthD-1 was used to assess T. pyriformis membrane damage during the 48-h experiment. The colorimetric MTT assay proved to be highly sensitive even at very short periods of Triton X-100 exposure. Tests performed in this study included simple and fast bioassays that provide overall information on the morphological and physiological state of cells exposed to different non-lytic and lytic concentrations of Triton X-100. PMID- 12781215 TI - Ara-C induces apoptosis in monkey fibroblast cells. AB - The effect of cytosine arabinoside (Ara-C) on cell viability has been studied in African green monkey kidney fibroblasts (CV1-P). It has been shown previously that Ara-C- induced cell death in neurons is mediated by apoptosis. We investigated whether Ara-C can induce apoptosis also in CV1-P cells, and if the apoptosis is p53-associated. For comparison, human neuroblastoma cells (SH-SY5Y) were studied as a model of human neuronal cells. SYTO13/propidium iodide staining revealed condensed and fragmented nuclei in both cell lines. Ara-C treatment for 48 h induced approximately 24% apoptosis in CV1-P cells whereas approximately 55% of SH-SY5Y cells were apoptotic. Ara-C increased the level of p53 in both CV1-P and SH-SY5Y cells compared to control. The maximum level of p53 in SH-SY5Y cells was reached at 12 h and this then rapidly faded whereas CV1-P cells p53 levels remained elevated after reaching their maximum. Caspase-3 activity was 5-fold higher in human neuroblastoma cells than in monkey fibroblasts, this reflected the decreased cell viability. Our results prove that Ara-C- induced apoptosis in CV1-P cells is associated with an increase of p53 and activation of caspase-3. Ara-C-induced toxicity in CV1-P cells is modest compared to that seen in neuronal cells. PMID- 12781216 TI - Forskolin-induced cyclic AMP signaling in single adherent bovine oviductal cells: effect of dichlorodiphenyltrichloroethane (DDT) and tris(4-chlorophenyl)methanol (TCPM). AB - The influence of tris(4-chlorophenyl)methanol (TCPM) and dichlorodiphenyltrichloroethane (o,p'DDT) on forskolin induced cAMP signalling in single adherent bovine oviductal cells was investigated. An increase in the intracellular cAMP levels was measured indirectly by an increase in the 520/580 nm fluorescence emission ratio of the protein kinase A fluorosensor (FICRhR). FICRhR was microinjected into single cells, and the 520/580 nm fluorescence emission ratio was monitored by image cytometry with an image analysis system as a measure of intracellular cAMP concentration ([cAMP](i)). Applications of dibutyryl cAMP and forskolin caused time- and dose-dependent effects on [cAMP](i) in single oviductal cells. The addition of 16 or 32 microM TCPM or DDT for 1 h to the culture medium decreased the intracellular cAMP concentration significantly, whereas 8 microM was not able to influence the [cAMP](i). In the presence of both pesticides at 16 microM the forskolin (30 microM)-induced [cAMP](i) was significantly reduced after 1 h of incubation. It is suggested that TCPM can have the same influence compared with DDT on cells responsible for reproduction. PMID- 12781217 TI - Release of salts from municipal solid waste combustion residues. AB - Residues from fluidized bed combustion of municipal solid waste were investigated with respect to their leaching behavior and possible extraction of salts. The total water extractable amounts of Na, K, Ca, Cl(-), Br(-), F(-) and SO(4)(2-) along with the total dissolved solids of bottom, hopper, cyclone and bag house filter ashes were determined. A simple multistage washing process (using water as the extraction medium) was tested in lab scale experiments. The effect of variations in parameters, such as water to ash weight ratio, contact time, temperature and number of extraction steps was investigated. The leaching behavior of untreated and washed cyclone and bag house filter ashes was evaluated by a two-step batch-leaching test, i.e. the CEN test. The ashes investigated in this study can be arranged according to their decreasing water extractable contents and total dissolved solids as follows: filter ash > cyclone ash > hopper ash > bottom ash. A triple extraction with water at liquid to solid ratio 2 and extraction time 5 min gave the best results for the extraction of Ca, Na, K, Cl( ) and SO(4)(2-) from the cyclone as well as from the filter ashes. The leached amounts of salts in the CEN test performed on the washed cyclone ash were considerably lower than the corresponding amounts released from the unwashed ash. Thus, the washed cyclone ash was made more stable with respect to salt leachability. On the other hand, large amounts of salts were leached from the washed filter ashes as well as from unwashed filter ashes. Therefore, it can be concluded that three stage water extraction is not a suitable stabilization method for this type of filter ashes. PMID- 12781218 TI - Removal of unburned carbon from municipal solid waste fly ash by column flotation. AB - Unburned carbon (UC) is the major source of organic contaminants in municipal solid waste (MSW) fly ash. So most organic contaminants can be removed by the removal of the UC from the MSW fly ash. In this paper, we first used a technique of column flotation to remove UC from MSW fly ash. The influences of column flotation parameters on the recovery efficiency of UC were systematically studied. It was found that the UC recovery efficiency was greatly influenced by the gas flow rate, pH value, collector kerosene's concentration and the types of fly ash. By optimizing the above parameters, we have successfully removed 61.2% of the UC from MSW fly ash having 5.24% UC content. The removal mechanism was well accounted for the kinetic theory of column flotation and surface-chemistry theory. The results indicate that the column flotation technique is effective in removing the UC from MSW fly ash, and show that there is a strong possibility for practical application of this technique in removing the organic contaminants from MSW fly ash. PMID- 12781219 TI - Solid state dehalogenation of PCBs in contaminated soil using NaBH4. AB - In this work we present the results of an experimental study on the abatement of polychlorinated biphenyl (PCBs) in contaminated soil using a high energy milling technique, that promotes a reaction only by impact between milling bodies. A sample of soil from a controlled landfill was treated with powdered NaBH(4) using two different hydride/soil ratios (5 and 2.5% w/w). The efficiency of the dehalogenation/hydrogenation reaction was studied as a function of the milling time (3.5 up to 30 h). After each run, the total PCBs content and the production of inorganic chloride were measured. The complete abatement was obtained with a starting PCBs concentration of about 2600 mg/kg. The residual PCBs concentration resulted to be <0.2 mg/kg. The final products of the treatment were biphenyl and NaCl. Other toxic or hazardous organic by-products were not generated. Boron was found as boric acid. PMID- 12781220 TI - Prediction of unconfined compressive strength of cement paste containing industrial wastes. AB - Neural network analysis was used to construct models of unconfined compressive strength (UCS) as a function of mix composition using existing data from literature studies of Portland cement containing real industrial wastes. The models were able to represent the known non-linear dependency of UCS on curing time and water content, and generalised from the literature data to find relationships between UCS and quantities of five waste types. Substantial decreases in UCS were caused by all wastes; except for EAF dust, the effect was nonlinear with the greatest decrease caused initially by approx. 12% plating sludge, 40% foundry dust, 58% other ash, and 72% MSWI fly ash by mass of dry product. It appears that the maximum waste additions used in modelling may approximate the practical limits of waste additions used in modelling may approximate the practical limits of waste addition to Portland cement, i.e., 50% plating sludge or EAF dust, 64% foundry dust, 92% other ash, and 85% MSWI fly ash by mass of dry product. The laboratory was found to be a key predictive variable and acted as a surrogate for laboratory-specific variables related to cement composition, strength and hardening class, product mixing and preparation details, laboratory conditions, and testing details. While the neural network modelling approach has been shown to be feasible, development of better models would require larger data sets with more complete information regarding laboratory-specific variables and waste composition. PMID- 12781221 TI - Wasteless combined aggregate-coal-fired steam-generator/melting-converter. AB - A method of reprocessing coal sludge and ash into granulate for the building industry in a combined wasteless aggregate-steam-generator/melting-converter was developed and tested. The method involves melting sludge and ash from coal-fired steam-generators of power plants in a melting-converter installed under the steam generator, with direct sludge drain from the steam generator combustion chamber. The direct drain of sludge into converter allows burnup of coal with high ash levels in the steam-generator without an additional source of ignition (natural gas, heating oil, etc.). Specific to the melting process is the use of a gas-air mixture with direct combustion inside a melt. This feature provides melt bubbling and helps to achieve maximum heat transfer from combustion products to the melt, to improve mixing, to increase rate of chemical reactions and to improve the conditions for burning the carbon residue from the sludge and ash. The "gross" thermal efficiency of the combined aggregate is about 93% and the converter capacity is about 18 t of melt in 100 min. The experimental data for different aspects of the proposed method are presented. The effective ash/charging materials feeding system is also discussed. The reprocessed coal ash and sludge in the form of granules can be used as fillers for concrete and as additives in the production of cement, bricks and other building materials. PMID- 12781222 TI - Water leaching of titanium from ore flotation residue. AB - Copper ore tailings were tested for the stability of titanium submitted to water leaching in three different reactor systems (agitated vessel, bioreactor and percolated fixed-bed column). For each of these systems, titanium extraction did not exceed 1% of the available metal. Biomass removed from ore residue adsorbed a small part of the titanium with sorption capacities below 20-30 mg g(-1), but most of this biomass was sequestered in the ore residue. Oxygen and carbon dioxide concentrations were monitored and changes in concentration correlated with bacteria development at the initial stage of the process and to fungal development in the latter stages. PMID- 12781223 TI - Preparation of activated carbons from heavy-oil fly ashes. AB - The use of heavy oil fly ash with high ash content (45 wt.%) as a precursor for the preparation of activated carbons has been investigated. The raw fly ash and the fly ash with lower ash content, obtained by a HCl/HF washing treatment, have been pyrolyzed at 900 degrees C and then activated with CO(2) in the temperature range of 800-900 degrees C for different times. The activated carbons have been characterised as regards the surface area and the pore volume. The evolution of the porosity has been related to the burn-off degree. PMID- 12781224 TI - The fixation and leaching of cement stabilized arsenic. AB - The solidification/stabilization of sodium arsenite, sodium arsenate, arsenic trioxide and arsenic pentoxide at dosages of approximately 10% has been investigated using sequential batch leaching tests. The leaching of arsenic, which was found to be diffusion based, was clearly least effective for those formulations containing additional iron(II). Calcium was found to influence the leaching of cement immobilized arsenic: those formulations containing the greatest Ca:As mole ratios were generally the most successful. Analysis using both FTIR and SEM revealed substantial changes to the cement matrices of those formulations to which the ferrous sulfate had been added. Ettringite was identified in the cement+ferrous sulfate formulations. PMID- 12781226 TI - Reuse of sewage sludge ashes (SSA) in cement mixtures: the effect of SSA on the workability of cement mortars. AB - The influence of sewage sludge ash (SSA) on workability of cement mortars has been studied. The irregular morphology of SSA particles produced a decrease of mortar workability. A nonlinear reduction of workability in mortars containing SSA was observed, but when SSA content in mortars was increased the workability reduction was less significant. A superplasticizer is used in order to compensate the decrease of workability produced by SSA. When SSA sized fractions were used, only significant differences in workability for mortars prepared with high water volumes or with the presence of superplasticizer were observed. PMID- 12781225 TI - Vitrification of lead-rich solid ashes from incineration of hazardous industrial wastes. AB - Lead-rich solid industrial wastes were vitrified by the addition of glass formers in various concentrations, to produce non-toxic vitreous stabilized products that can be freely disposed or used as construction materials. Toxicity of both the as received industrial solid waste and the stabilized products was determined using standard leaching test procedures. The chemically stable vitreous products were subjected to thermal annealing in order to investigate the extent of crystal separation that could occur during cooling of large pieces of glass. Leaching tests were repeated to investigate the relation between annealing process and chemical stability. X-ray, scanning and transmission electron microscopy techniques were employed to identify the microstructure of stabilized products before and after thermal treatment. Relation between synthesis and processing, chemical stability and microstructure was investigated. PMID- 12781227 TI - Wet air oxidation of a reactive dye solution using CoAlPO(4)-5 and CeO(2) catalysts. AB - Wet air oxidation of a prepared reactive dye solution was performed to assess the efficacy of CoAlPO(4)-5 and CeO(2) as catalysts in the reaction. Via adsorption and oxidation of dye, CoAlPO(4)-5 effectively decreased American Dye Manufacturers Institute and chemical oxygen demand (COD) values in the dye solution. At a reaction temperature of 135 degrees C and an applied pressure of 1.0 MPa, color and COD removal were as high as 95% and 90%, respectively, after 2 h. Active sites on the outer surface of CoAlPO(4)-5 are responsible for adsorption and decomposition of dye while active sites in the pores dominate further destruction and oxidation of intermediate products. Since the outer surface only represents a minor part of the total surface, the color removal does not increase appreciably with loading of CoAlPO(4)-5. The CeO(2) catalyst, calcined from cerium chloride under high thermal impact (type A CeO(2)) was very effective in removing color and COD from the solution. This catalyst demonstrated near 100% color removal at temperatures above 135 degrees C and the COD removal could be above 95% at 165 degrees C. With both CoAlPO(4)-5 and CeO(2) catalysts, COD rose and then fell back during the reaction, a feature typical of a consecutive reaction. In contrast to prepared CeO(2), a commercial CeO(2) did not exhibit any catalytic ability for the removal of color and COD. The durability of both CoAlPO(4)-5 and prepared CeO(2) is considered to be fair. PMID- 12781228 TI - CO(2)-H(2)-dependent anaerobic biotransformation of phthalic acid isomers in sediment slurries. AB - This paper investigates the anaerobic biotransformation of three isomers of phthalic acid and benzoic acid in sediment slurries under four different atmospheres [N(2), N(2)/H(2) (19:1, v/v), CO(2), and CO(2)/H(2) (4:1, v/v)]. Significant differences were observed in lag periods and biotransformation rates among the phthalic acid isomers and under the different atmospheres. In most cases, the relative biotransformation rates of the three isomers of phthalic acid were ortho-phthalic acid>isophthalic acid>terephthalic acid. Benzoate was transformed faster than any isomer of phthalic acid. Since biotransformation of phthalic acid isomers in sediment slurries was enhanced by high initial levels of H(2) and CO(2) in the headspace, we propose a pathway for phthalic acid biodegradation in which the initial transformation to benzoate is CO(2)-H(2) dependent. Acetogenic bacteria were investigated for their possible involvement in this transformation reaction, but when MPN counts were used to compare the growth dynamics of acetogenic bacteria with the time course of the terephthalic acid transformation under N(2)/H(2) (19:1, v/v) and CO(2)/H(2) (4:1, v/v) atmospheres, the results were inconclusive. PMID- 12781229 TI - Characterization of lead, cadmium, arsenic and nickel in PM(2.5) particles in the Athens atmosphere, Greece. AB - Concentrations of Pb, Cd, As and Ni in PM(2.5) particles were measured in samples collected, using low volume PM(2.5) samplers (Harvard Impactor system, HI) at two sites in Athens basin; Patission Street in Athens city center and Rentis, a semi urban and industrial area, during March 1995-March 1996. Sample analysis for Pb, Cd, Ni and As was accomplished by electrothermal atomic absorption spectrometry after total digestion. Annual geometric mean values in 183 PM(2.5) particles samples were found to be: Pb: 143 nanogram(-3); Cd: 0.34 nanogram(-3); Ni: 4.55 nanogram(-3); As: 0.79 nanogram(-3). The geographical and temporal distribution patterns were investigated. Pb exhibited higher values during the winter period. For the other elements no significant seasonal variation was observed. Wind direction, air temperature and relative humidity affected element concentrations. Principal component analysis was applied on the data to enable source apportionment of toxic elements in PM(2.5) particles. It was found that Pb, As and Ni have common sources, which could be vehicles emissions/oil combustion and resuspended road dust. Cd and a portion of As originate from industrial activities. PMID- 12781230 TI - Comparative studies of fungal degradation of single or mixed bioaccessible reactive azo dyes. AB - A screening using several fungi (Phanerochaete chrysosporium, Pleurotus ostreatus, Trametes versicolor and Aureobasidium pullulans) was performed on the degradation of syringol derivatives of azo dyes possessing either carboxylic or sulphonic groups, under optimized conditions previously established by us. T. versicolor showed the best biodegradation performance and its potential was confirmed by the degradation of differently substituted fungal bioaccessible dyes. Enzymatic assays (lignin peroxidase, manganese peroxidase, laccase, proteases and glyoxal oxidase) and GC-MS analysis were performed upon the assay obtained using the most degraded dye. The identification of hydroxylated metabolites allowed us to propose a possible metabolic pathway. Biodegradation assays using mixtures of these bioaccessible dyes were performed to evaluate the possibility of a fungal wastewater treatment for textile industries. PMID- 12781231 TI - Effects of temperature on mineralisation of petroleum in contaminated Antarctic terrestrial sediments. AB - Although petroleum contamination has been identified at many Antarctic research stations, and is recognized as posing a significant threat to the Antarctic environment, full-scale in situ remediation has not yet been used in Antarctica. This is partly because it has been assumed that temperatures are too low for effective biodegradation. To test this, the effects of temperature on the hydrocarbon mineralisation rate in Antarctic terrestrial sediments were quantified. 14C-labelled octadecane was added to nutrient amended microcosms that were incubated over a range of temperatures between -2 and 42 degrees C. We found a positive correlation between temperature and mineralisation rate, with the fastest rates occurring in samples incubated at the highest temperatures. At temperatures below or near the freezing point of water there was a virtual absence of mineralisation. High temperatures (37 and 42 degrees C) and the temperatures just above the freezing point of water (4 degrees C) showed an initial mineralisation lag period, then a sharp increase in the mineralisation rate before a protracted plateau phase. Mineralisation at temperatures between 10 and 28 degrees C had no initial lag phase. The high rate of mineralisation at 37 and 42 degrees C was surprising, as most continental Antarctic microorganisms described thus far have an optimal temperature for growth of between 20 and 30 degrees C and a maximal growth temperature <37 degrees C. The main implications for bioremediation in Antarctica from this study are that a high-temperature treatment would yield the most rapid biodegradation of the contaminant. However, in situ biodegradation using nutrients and other amendments is still possible at soil temperatures that occur naturally in summer at the Antarctic site we studies (Casey Station 66 degrees 17(') S, 110 degrees 32(') E), although treatment times could be excessively long. PMID- 12781232 TI - Removal of ammonia solutions used in catalytic wet oxidation processes. AB - Ammonia (NH(3)) is an important product used in the chemical industry, and is common place in industrial wastewater. Industrial wastewater containing ammonia is generally either toxic or has concentrations or temperatures such that direct biological treatment is unfeasible. This investigation used aqueous solutions containing more of ammonia for catalytic liquid-phase oxidation in a trickle-bed reactor (TBR) based on Cu/La/Ce composite catalysts, prepared by co-precipitation of Cu(NO(3))(2), La(NO(3))(2), and Ce(NO(3))(3) at 7:2:1 molar concentrations. The experimental results indicated that the ammonia conversion of the wet oxidation in the presence of the Cu/La/Ce composite catalysts was determined by the Cu/La/Ce catalyst. Minimal ammonia was removed from the solution by the wet oxidation in the absence of any catalyst, while approximately 91% ammonia removal was achieved by wet oxidation over the Cu/La/Ce catalyst at 230 degrees C with oxygen partial pressure of 2.0 MPa. Furthermore, the effluent streams were conducted at a liquid hourly space velocity of under 9 h(-1) in the wet catalytic processes, and a reaction pathway was found linking the oxidizing ammonia to nitric oxide, nitrogen and water. The solution contained by-products, including nitrates and nitrites. Nitrite selectivity was minimized and ammonia removal maximized when the feed ammonia solution had a pH of around 12.0. PMID- 12781233 TI - Treatment of coke-plant wastewater by biofilm systems for removal of organic compounds and nitrogen. AB - Coke-plant wastewater was treated by an anaerobic-anoxic-aerobic (A(1)-A(2)-O) biofilm system and an anoxic-aerobic (A/O) biofilm system, respectively. At same or similar levels of hydraulic retention time (HRT), the two systems had almost identical chemical oxygen demand (COD) and NH(3) removals, but a different organic-N removal. Set-up of an acidogenic stage benefited for the removal of organic-N and the A(1)-A(2)-O system was more useful for total nitrogen removal than the A-O system. HRT did not have a substantial effect on the COD and NH(3)-N removal efficiencies, but considerably influenced the organic-N removal and distribution of oxidized nitrogen in the final effluent. The GC/MS analysis demonstrated that some refractory compounds were decomposed at the acidogenic stage and resulted in the production of some intermediates, which were more readily degraded in the subsequent aerobic stage. Hence, the A(1)-A(2)-O system had better effluent quality than the A-O system in terms of effluent composition. PMID- 12781234 TI - Indoor and outdoor carbon monoxide concentration relationships at different microenvironments in the Athens area. AB - Simultaneous measurements of indoor and outdoor carbon monoxide (CO) concentrations conducted at two different microenvironments in Athens, Greece, using a non-dispersive infrared analyzer, are described in this paper. The two selected microenvironments, an office and a public school, were located in the vicinity of two streets with heavy traffic, near the center of Athens. A statistical correlation analysis of indoor concentration levels with outdoor concentrations monitored at the school and at the office, as well as with meteorological parameters and outdoor concentrations monitored at a fixed monitoring site, was conducted. Hourly indoor concentrations at the office and at the school showed a significant positive correlation with outdoor concentrations measured at both measurement locations (with correlation coefficient values R=0.74 and R=0.83 respectively) and at the fixed site (with R=0.70 and R=0.67 respectively). The correlation between indoor and outdoor concentrations was even better when hourly concentrations averaged over a 4 h period were considered (correlation coefficient values between indoor and outdoor concentrations measured at the office and at the school were R=0.85 and R=0.92 respectively and the correlation coefficient value between indoor and outdoor fixed site concentrations was R=0.75 for both sites). Mean hourly outdoor concentrations at the fixed monitoring site explained approximately 56% (R=0.75) of the variation of outdoor concentrations at the office and approximately 68 % (R=0.83) of the variation of outdoor concentrations at the school. The mean daily indoor to outdoor (I/O) ratio ranged between 0.74 and 1.00 at the office and between 0.53 and 0.89 at the school. PMID- 12781235 TI - Equilibrium studies for the sorption of lead from effluents using chitosan. AB - The sorption of lead ions from aqueous solution onto chitosan has been studied. Equilibrium studies have been carried out to determine the capacity of chitosan for lead ions. The effects of solution pH and chitosan particle size on the sorption capacity have been studied.The experimental data were analyzed using three equilibrium isotherm correlations, namely, Langmuir, Freundlich and Redlich Peterson equations. The linear correlation coefficients were determined for each isotherm and the Freundlich provided the best fit. In addition, error functions have been used to determine the alternative single component equilibrium isotherm parameters by non-linear regression due to the inherent bias in using the correlation coefficient from the linearization. This technique enables the "best fit" isotherm parameters to be used in the equilibrium equations for the sorption of lead ions on chitosan within the limits and assumptions of the various error analysis methods. PMID- 12781237 TI - Noble metal catalysts for methane removal. AB - Using a bench-scale rig, the activities of Pt, Pd and Pt+Pd catalysts supported on gamma-Al(2)O(3) and on TiO(2) (anatase) for the complete oxidation of methane (300 ppmv) in air have been measured as a function of temperature; values of T(10), T(50) and T(90) together with the Arrhenius parameters (activation energy and pre-exponential factor) are reported. Pt is less active than Pd when deposited on the surface of the TiO(2), but more active when deposited on gamma Al(2)O(3), however when combined, the Pt+Pd mixture is more active than either metal individually. The T(10) for Pt+Pd/gamma-Al(2)O(3) was being as low as 228 degrees C. The significance of the Arrhenius parameters, for metal containing catalysts is that they exhibit compensation with increasing activation energy, while securing a more rapid increase in conversion from 0% to 100% when the temperature is increased. PMID- 12781238 TI - Toxic by-products from the combustion of Kraft lignin. AB - Lignin samples, sub-product in the Kraft process of cellulose from eucalyptus wood, were burnt in a laboratory scale furnace at different residence temperatures and with distinct fuel-rich atmospheres. The yields of CO, CO(2), eight light hydrocarbons (methane, ethylene, ethane, propylene, acetylene, butane, etc.) and 60 semi-volatile+volatile compounds (benzene, toluene, ethylbenzene, styrene, indene, naphthalene, dibenzofuran, phenanthrene, chrysene, etc.) were determined, with nominal reactor temperatures between 800 and 1100 degrees C and residence times of the volatiles evolved and formed between 4 and 7 s. The collection of the gases and volatiles evolved was carried out with a Tedlar bag and by XAD-4 resin respectively, comparing the data obtained in both cases. The emission factor (mg/kg) of the CO was between 2500 and 90000, and under the poor-oxygen atmosphere, the emission factors of many by-toxic products were greater than 100 mg/kg. A pyrolysis run was also performed, obtaining emission factors between 30 and 3000 mg/kg, facilitating its identification. The behaviour of different compounds in the combustion runs was discussed considering three groups in accordance with their stability vs. oxygen, and two groups vs. temperature. PMID- 12781236 TI - Growth conditions, elemental accumulation and induced physiological changes in Chinese cabbage. AB - Soils contaminated with low levels of heavy metals and other trace elements are now frequently used for vegetable growing. In this situation, heavy metals and trace elements from these polluted soils may accumulate in the agricultural plants being grown in them and thereby enter the human food chain. The objectives of this study are to elucidate the effects of growth conditions, manipulated by the crop covers, on the phytoaccumulation of elements, and to investigate the conceivable influences of these conditions on the plant biochemistry. In three consecutive years of field experiments, open air (T(0)), and floating rowcover treatments (T(1): perforated polyethylene 50 micrometers; T(2): polypropylene 17 gm(-2)) were used to produce different environmental conditions for the growth of Chinese cabbage [Brassica rapa L. (Pekinensis group) cv. 'Nagaoka 50']. Five samplings (whole tops) were carried out from transplanting to harvest and measurements of B, Al, Ag, Si and Ca concentration as well as phenolics (orto diphenols, total phenols and anthocyanins), pectic fractions, amino acids (histidine, phenylalanine and tyrosine) and polyphenol oxidase activity, were carried out in samples. The T(1) (perforated polyethylene sheet) gave greater B, Al, Ag and Si concentration and phytoextraction (in weight units) than the open air control. These findings can help to develop new cost-effective techniques for phytoremediation as the application of plastic covers in the field. The build-up of heavy metals in those crops would make the product less suitable for human consumption. PMID- 12781239 TI - Chromium species behaviour in the activated sludge process. AB - The purpose of this research was to compare trivalent chromium (Cr(III)) and hexavalent chromium (Cr(VI)) removal by activated sludge and to investigate whether Cr(VI) reduction and/or Cr(III) oxidation occurs in a wastewater treatment system. Chromium removal by sludge harvested from sequencing batch reactors, determined by a series of batch experiments, generally followed a Freundlich isotherm model. Almost 90% of Cr(III) was adsorbed on the suspended solids while the rest was precipitated at pH 7.0. On the contrary, removal of Cr(VI) was minor and did not exceed 15% in all experiments under the same conditions. Increase of sludge age reduces Cr(III) removal, possibly because of Cr(III) sorption on slime polymers. Moreover, the decrease of suspended solids concentration and the acclimatization of biomass to Cr(VI) reduced the removal efficiency of Cr(III). Batch experiments showed that Cr(III) cannot be oxidized to Cr(VI) by activated sludge. On the contrary, Cr(VI) reduction is possible and is affected mainly by the initial concentration of organic substrate, which acts as electron donor for Cr(VI) reduction. Initial organic substrate concentration equal to or higher than 1000 mgl(-1) chemical oxygen demand permitted the nearly complete reduction of 5 mgl(-1) Cr(VI) in a 24-h batch experiment. Moreover, higher Cr(VI) reduction rates were obtained with higher Cr(VI) initial concentrations, expressed in mg Cr(VI) g(-1) VSS, while decrease of suspended solids concentration enhanced the specific Cr(VI) reduction rate. PMID- 12781240 TI - Mechanism and kinetic model for the decolorization of the azo dye Reactive Black 5 by hydrogen peroxide and UV radiation. AB - A kinetic model for the decolorization of C.I. Reactive Black 5 by the combination of hydrogen peroxide and UV radiation was developed based on experimental results and known chemical and photochemical reactions. The observed kinetic reaction coefficient was determined and correlated as a function of hydrogen peroxide concentration and UV intensity. The validity of the rate expression was tested experimentally in a parameterization study. The decolorization rate follows pseudo-first order kinetics with respect to dye concentration. The rate increases linearly with UV intensity and nonlinearly with increasing hydrogen peroxide concentration, going from a linear relationship at low H(2)O(2) concentrations to a maximum as hydrogen peroxide concentration continues to increase. The decolorization rate expression derived from the proposed reaction mechanism was reconciled with that used for correlating the experimental data. PMID- 12781241 TI - Atmospheric lead concentration distribution in Northern Taiwan. AB - Atmospheric lead concentrations were measured randomly, approximately once per week, at five traffic sites in northern Taiwan from September 1994 to May 1995. Three types of theoretical distributions, lognormal, Weibull and gamma were selected to fit the frequency distribution of the measured lead concentration. Four goodness-of-fit criteria were used to judge which theoretical distribution is the most appropriate to represent the frequency distributions of atmospheric lead. The results show that atmospheric lead concentrations in total suspended particulates fit the lognormal distribution reasonably well in northern Taiwan. The intervals of fitted theoretical cumulative frequency distributions (CFDs) can successfully contain the measured data when the population mean is estimated with a 95% confidence interval. In addition, atmospheric lead concentration exceeding a critical concentration is also predicted from the fitted theoretical CFDs. PMID- 12781242 TI - Leaching behavior and chemical stability of copper butyl xanthate complex under acidic conditions. AB - Although xanthate addition can be used for treating copper-containing wastewater, a better understanding of the leaching toxicity and the stability characteristics of the copper xanthate complexes formed is essential. This work was undertaken to evaluate the leaching behavior of copper xanthate complex precipitates by means of toxicity characteristics leaching procedure (TCLP) and semi-dynamic leaching test (SDLT) using 1 N acetic acid solution as the leachant. Also, the chemical stability of the copper xanthate complex during extraction has been examined with the studying of variation of chemical structure using UV-vis, Fourier transform infrared and X-ray photoelectron spectroscopies (XPS). Both TCLP and SDLT results showed that a negligible amount of copper ion was leached out from the copper xanthate complex precipitate, indicating that the complex exhibited a high degree of copper leaching stability under acidic conditions. Nevertheless, chemical structure of the copper xanthate complex precipitate varied during the leaching tests. XPS data suggested that the copper xanthate complex initially contained both cupric and cuprous xanthate, but the unstable cupric xanthate change to the cuprous form after acid extraction, indicating the cuprous xanthate to be the final stabilizing structure. Despite that, the changes of chemical structure did not induce the rapid leaching of copper from the copper xanthate complex. PMID- 12781243 TI - Cultural abnormal psychology. PMID- 12781244 TI - Assessment of psychopathology across and within cultures: issues and findings. AB - Research based information on the impact of culture on psychopathology is reviewed, with particular reference to depression, somatization, schizophrenia, anxiety, and dissociation. A number of worldwide constants in the incidence and mode of expression of psychological disorders are identified, especially in relation to schizophrenia and depression. The scope of variation of psychopathological manifestations across cultures is impressive. Two tasks for future investigations involve the determination of the generic relationship between psychological disturbance and culture and the specification of links between cultural characteristics and psychopathology. To this end, hypotheses are advanced pertaining to the cultural dimensions investigated by Hofstede and their possible reflection in psychiatric symptomatology. It is concluded that the interrelationship of culture and psychopathology should be studied in context and that observer, institution, and community variables should be investigated together with the person's experience of distress and disability. PMID- 12781245 TI - Cultural differences in emotions: a context for interpreting emotional experiences. AB - In this article, it is suggested that cross-cultural assessment of emotional disturbances would benefit from the consideration of cultural differences in the modal, and normative emotions. A summary of the research literature on cultural differences in emotions, in particular in antecedent events, subjective feeling, appraisal, and behavior is provided. Cultural differences in emotions are understood functionally, such that the most prevalent emotional phenomena in a culture are those that fit and reinforce the distinct cultural models (i.e. goals and practices) of self and relationship. It is argued that a culture-sensitive approach to emotional disturbances would entail the assessment of emotional phenomena that are dysfunctional to the cultural models of self and relationship. PMID- 12781246 TI - Masculinity-femininity as a national characteristic and its relationship with national agoraphobic fear levels: Fodor's sex role hypothesis revitalized. AB - Hofstede's dimension of national culture termed Masculinity-Femininity [. Cultures and organizations: software of the mind. London: McGraw-Hill] is proposed to be of relevance for understanding national-level differences in self assessed agoraphobic fears. This prediction is based on the classical work of Fodor [. In: V. Franks & V. Burtle (Eds.), Women in therapy: new psychotherapies for a changing society. New York: Brunner/Mazel]. A unique data set comprising 11 countries (total N=5491 students) provided the opportunity of scrutinizing this issue. It was hypothesized and found that national Masculinity (the degree to which cultures delineate sex roles, with masculine or tough societies making clearer differentiations between the sexes than feminine or modest societies do) would correlate positively with national agoraphobic fear levels (as assessed with the Fear Survey Schedule-III). Following the correction for sex and age differences across national samples, a significant and large effect-sized national-level (ecological) r=+0.67 (P=0.01) was found. A highly feminine society such as Sweden had the lowest, whereas the champion among the masculine societies, Japan, had the highest national Agoraphobic fear score. PMID- 12781247 TI - Higher levels of state depression in masculine than in feminine nations. AB - Studies using identical measures have identified different levels of depression in different countries or cultures. Until now, however, explanations for such differences, other than methodological ones, have not been empirically addressed. It was hypothesized and found that soft or feminine nations in which both women and men are offered equal opportunities for the fulfillment of multiple social roles that are associated with good self-rated health would score significantly lower on national depression levels than tough or masculine societies in which such opportunities exist to a clearly lesser extent. Analyses of data collected in 14 nations in Europe (total N>5000) demonstrated that higher scores on Hofstede's national masculinity index and lower ones on national wealth were independent predictors of higher national depression levels. National trait neuroticism did not mediate the relationship between national masculinity and national depression levels. PMID- 12781248 TI - Objective personality assessment of people in diverse cultures: European adaptations of the MMPI-2. AB - This article examines the cross-national application of psychological tests and examines the generalizability of objective psychological assessment instruments. The most widely used and internationally adapted personality instrument, the Minnesota Multiphasic Personality Inventory (MMPI-2), is highlighted to illustrate the adaptation of psychological tests across the barriers of language and culture. The problems and limitations of using questionnaire methods were noted and effective strategies for translating, adapting, and standardizing questionnaires in languages and cultures different from their country of origin are reviewed. The history of several European adaptations of the original MMPI and MMPI-2 is surveyed to illustrate the extensive research base for the test in Europe. Applications of the MMPI-2 in clinical and non-clinical (e.g. industrial) settings were noted. Current research is described and recommendations for future research are provided. PMID- 12781251 TI - Emergence of the role of cataract and IOL surgery in the correction of refractive errors. PMID- 12781249 TI - Anxiety sensitivity in six countries. AB - In the present study, the Anxiety Sensitivity Index-Revised (ASI-R; ) was administered to a large sample of persons (n=2786) from different cultures represented in six different countries: Canada, France, Mexico, The Netherlands, Spain, and the United States. We sought to (a) determine the factor structure and internal consistency of the ASI-R and (b) examine the correlations of the measure with psychiatric symptoms and personality dimensions in a single European non English speaking country (The Netherlands). Partially consistent with the original hypothesis, the underlying structure of the anxiety sensitivity construct was generally similar across countries, tapping fear about the negative consequences of anxiety-related physical and social-cognitive sensations. Lower order factors were moderately to strongly correlated with one another and showed good internal consistency. The observed lower-order ASI-R factors correlated with established psychiatric symptoms and with the personality trait of neuroticism. Partial correlations indicated that both factors are useful in accounting for variance in symptom measures. We discuss the results of this investigation in relation to the cross-cultural assessment of the anxiety sensitivity construct. PMID- 12781252 TI - Lamellar or surface ablation? PMID- 12781254 TI - Array multifocal IOLs in a charity hospital training program. PMID- 12781256 TI - Posttraumatic neovascularization in a cataractous crystalline lens. PMID- 12781258 TI - Trypan-blue-assisted capsulorhexis for trainee phacoemulsification surgeons. PMID- 12781260 TI - Consultation section: refractive surgical problem. PMID- 12781259 TI - Posttraumatic neovascularization. PMID- 12781268 TI - Laser in situ keratomileusis and intrastromal corneal ring segments for high myopia. Three-step procedure. AB - We describe a surgical procedure to correct relatively high myopia in patients with thin corneas. The technique combines laser in situ keratomileusis (LASIK) and intrastromal corneal ring segment (ICRS) implantation. The 3-step procedure comprises ICRS channel formation, LASIK, and segment insertion. The technique was used in 2 eyes of 2 patients with low pachymetry values and high myopia. One patient achieved an uncorrected visual acuity (UCVA) of 20/30 and a best spectacle-corrected visual acuity (BSCVA) of 20/25 with +0.50 -1.75 x 101. The other patient achieved a UCVA of 20/30 and a BSCVA of 20/20 with +0.75 -1.50 x 80. PMID- 12781269 TI - Sutureless ciliary sulcus supported intraocular lens with transiridal anchoring haptics. AB - We report a technique for implanting intraocular lenses (IOLs) in the ciliary sulcus in eyes without capsule support. The IOL design allows it to be implanted without fixation sutures. The lens was implanted in 3 aphakic eyes without capsule support. PMID- 12781270 TI - Laser in situ keratomileusis for consecutive hyperopia after myopic LASIK and radial keratotomy. AB - PURPOSE: To evaluate and compare the efficacy, predictability, and safety of hyperopic laser in situ keratomileusis (H-LASIK) for the correction of consecutive hyperopia after myopic-LASIK (M-LASIK) and radial keratotomy (RK). SETTING: The Eye Institute of Utah, Salt Lake City, Utah, USA. METHODS: Seventy seven eyes of 64 patients were studied. The eyes were divided into 2 groups based on the prior refractive procedures: in Group A (n = 34), H-LASIK was performed for overcorrection after M-LASIK and in Group B (n = 43), for overcorrection after RK. All eyes were included in the analysis of intraoperative and postoperative complications. Only eyes with a minimum follow-up of 6 months were included in the analysis of visual and refractive results. Among these 66 eyes, 30 were in Group A and 36 were in Group B. The mean follow-up in these eyes was 12.34 months +/- 5.95 (SD) (range 6 to 33 months). RESULTS: Overall, the mean spherical equivalent (SE) was +1.88 +/- 0.91 diopters (D) preoperatively and 0.37 +/- 0.65 D at the last visit. Eighty-three percent of eyes were within +/ 1.00 D of emmetropia, and 66% were within +/-0.50 D. The uncorrected visual acuity (UCVA) was 20/20 in 39% of eyes and 20/40 or better in 92% of eyes. The preoperative SE was +1.43 +/- 0.59 D in Group A and +2.26 +/- 0.96 D in Group B; the difference in the preoperative SE was significant (P=.001). However, there was no statistically significant between-group difference in postoperative refraction and UCVA. One eye in Group B (3%) lost 2 or more lines of best corrected visual acuity. Corneal ectasia developed in 1 eye in Group B 11 months after H-LASIK. A sliver occurred in 1 eye in Group A after the flap was recut. CONCLUSION: Hyperopic LASIK was equally effective and predictable in treating consecutive hyperopia after overcorrected M-LASIK and overcorrected RK. The safety of the procedure in the RK group appeared to be inferior to that in the M LASIK group. Although vision-threatening complications are rare after H-LASIK retreatment, corneal ectasia developed in 1 eye in the RK group. PMID- 12781271 TI - Photorefractive keratectomy for the treatment of purely refractive accommodative esotropia. AB - PURPOSE: To evaluate the results of photorefractive keratectomy (PRK) for the treatment of young adult patients with purely refractive accommodative esotropia. SETTING: Private practice and university hospital, Milan, Italy. METHODS: The medical records of consecutive patients who had PRK for hyperopia associated with purely refractive esotropia were reviewed retrospectively. Preoperative and postoperative visual acuity, alignment, and sensory data were recorded and analyzed. Surgical methods and complications were reviewed. RESULTS: Sixteen eyes of 8 patients were treated. The mean patient age at the time of treatment was 24.6 years (range 17 to 38 years). All patients were followed for 1 year. At the 1-year follow-up evaluation, the uncorrected visual acuity was 20/40 or better in all eyes. No patient lost a line of best spectacle-corrected visual acuity. The mean spherical equivalent was -3.7 diopters (D) preoperatively and -0.7 D postoperatively. All patients were within +/-0.37 D of emmetropia at the 1-year evaluation. Preoperatively, the mean esotropic deviation was 10.75 prism diopters. Postoperatively, all patients were orthophoric without correction. Stereopsis was unaffected by PRK in all patients. There were no intraoperative or postoperative complications. CONCLUSION: Photorefractive keratectomy was an effective treatment for esotropia associated with mild to moderate hyperopia in young adults with purely refractive accommodative esotropia. These findings should not be widely applied to children with accommodative esotropia. PMID- 12781272 TI - Clear lens extraction with intraocular lens implantation for hyperopia. AB - PURPOSE: To analyze the results of clear lens extraction (CLE) with posterior chamber intraocular lens (IOL) implantation to correct hyperopia. SETTING: Eye Research Center and Dr. Agarwal's Eye Hospital, Chennai, India. METHODS: This prospective study comprised 20 hyperopic eyes of 12 patients between 19 and 50 years who had CLE with posterior chamber IOL implantation. Five patients had peripheral iridectomy during CLE as the angles were occludable. RESULTS: The mean hyperopic spherical equivalent refraction was +6.66 diopters (D) +/- 2.17 (SD) (range +4.75 to +13.00 D). The IOL power was calculated using the Holladay 2 formula. The mean follow-up was 16.96 months (range 6 to 35 months). The mean postoperative uncorrected visual acuity was 0.45 +/- 0.25 (range 0.10 to 1.00), a mean improvement of 3 Snellen lines from preoperatively. The mean postoperative best corrected visual acuity (BCVA) was 0.63 +/- 0.30, a mean improvement of 1 Snellen line. Three patients gained 2 lines of BCVA and 2 patients, 1 line. One patient lost 1 line of BCVA. Seventy percent of patients were within +/-0.50 D of the intended refraction. CONCLUSION: The results indicate that CLE with posterior chamber IOL implantation is safe, predictable, and effective. PMID- 12781273 TI - Comparison of photorefractive keratectomy and laser in situ keratomileusis for the treatment of compound hyperopic astigmatism. AB - PURPOSE: To compare photorefractive keratectomy (PRK) and laser in situ keratomileusis (LASIK) for compound hyperopic astigmatism. SETTING: University laser center. METHODS: This prospective nonrandomized study evaluated 41 consecutive eyes (27 patients) that had PRK and 24 consecutive eyes (15 patients) that had LASIK to correct compound hyperopic astigmatism. RESULTS: The mean preoperative error was +3.06 diopters of sphere (DS) +/-1.73 (SD)/+1.31 +/- 0.60 diopters of cylinder (DC) in the PRK eyes and +2.86 +/-1.28 DS/+1.55 +/- 0.96 DC in the LASIK eyes. The mean maximal pain score in PRK eyes was 1.95 +/- 1.19 (range 0.0 to 3.0) in PRK eyes and 0.84 +/-1.12 in LASIK eyes (P=.0014). The uncorrected visual acuity was 20/20 or better in 7.7% of the PRK eyes and 58.3% of the LASIK eyes at 1 month (P<.001) and 57.9% and 66.7%, respectively, at 9 months (P=.586). The mean postoperative spherical error was -0.95 +/- 0.92 D in PRK eyes and +0.33 +/- 0.56 D in LASIK eyes at 1 month (P<.001) and +0.64 +/- 1.01 D and +0.44 +/- 0.57 D, respectively, at 9 months (P=.375). There was no statistically significant between-group difference in the mean residual astigmatic error. Mild peripheral haze (grade 0.5 to 1.0) occurred in 19.5% of PRK eyes and no LASIK eye. No eye in either group lost more than 2 lines of best spectacle-corrected visual acuity. CONCLUSIONS: Photorefractive keratectomy was more painful than LASIK and led to a slower visual recovery, a higher incidence of peripheral haze, and an initial myopic overcorrection, which self-corrected by 3 to 6 months. Efficacy and stability of the astigmatic correction were similar in both groups. Long-term stability of both procedures requires further study. PMID- 12781274 TI - Combined laser in situ keratomileusis and radial keratotomy for the treatment of moderate to high myopia. AB - PURPOSE: To evaluate the results of combining laser in situ keratomileusis (LASIK) and radial keratotomy (RK) for the correction of moderate to high myopia. SETTING: Corneal Consultants of Colorado, Littleton, Colorado, and International Eye Care Laser Vision Center, Houston, Texas, USA. METHODS: In a retrospective 2 surgeon interventional case series, outcomes in 60 eyes of 41 patients with moderate to high myopia were reported. All eyes had LASIK using a Visx Star S2 or Summit Apex Plus((R)) excimer laser. Residual myopia was treated with RK. RESULTS: Of the 54 eyes corrected for distance, the mean refractive spherical equivalent was -8.09 diopters (D) preoperatively and -0.43 D after LASIK and RK. The mean follow-up was 15.4 months. At the last visit, the uncorrected visual acuity was 20/20 in 22 eyes (41%), 20/25 in 36 eyes (67%), and 20/40 in 51 eyes (94%). No eye lost 2 or more lines of best spectacle-corrected visual acuity. CONCLUSIONS: Many patients previously considered poor refractive surgery candidates because of their degree of myopia or corneal thickness could be offered this safe and effective combined refractive technique. PMID- 12781275 TI - Treatment of simple hyperopia: comparison of laser in situ keratomileusis and laser thermal keratoplasty. AB - PURPOSE: To evaluate and compare the efficacy, stability, and safety of laser in situ keratomileusis (LASIK) and laser thermal keratoplasty (LTK) for the treatment of simple hyperopia. SETTING: John Hill Eye and Laser Centre, Cape Town, South Africa. METHODS: This retrospective study comprised consecutive patients having primary treatment of simple hyperopia of up to 3.0 diopters (D) with astigmatism of 0.5 D or less. Treatment methods were as follows: Group 1 (81 eyes), LASIK with the Nidek EC-5000 excimer laser; Group 2 (69 eyes), LASIK with the LaserSight LSX excimer laser; and Group 3 (84 eyes), LTK with the Sunrise holmium:YAG laser. RESULTS: The hyperopia decreased in all 3 groups. Both LASIK groups were stable by 1 month; there was continued regression in the LTK group for up to 18 months. The percentage of eyes achieving uncorrected visual acuities of 20/20 and 20/40 at 3 months were Nidek, 41% and 92%, respectively; LaserSight, 50% and 90%, respectively; and LTK, 21% and 89%, respectively. No eye lost more than 2 lines of best spectacle-corrected visual acuity. Surgically induced astigmatism (SIA) was evident in all 3 groups; it was highest in the LTK group. The mean posttreatment astigmatism was -0.47 D +/- 0.40 (SD) (range 0 to -1.50 D) in the Nidek group, -0.45 +/- 0.40 D (range 0 to -1.25 D) in the LaserSight group, and -0.81 +/- 0.51 D (range 0 to -2.25 D) in the LTK group. The enhancement rates were 16.75%, 22.57%, and 38.30%, respectively. Because of the SIA, 61% of the LTK enhancements were corrected with LASIK. CONCLUSIONS: All 3 treatment methods corrected hyperopia, but stability was achieved early in both LASIK groups, allowing early enhancement when necessary. Because LTK cannot currently correct astigmatism, many of the LTK repeat treatments required LASIK procedures. For these reasons, LASIK remains my preferred method to treat simple hyperopia up to +3.0 D. PMID- 12781276 TI - Implantable contact lens for moderate to high myopia: relationship of vaulting to cataract formation. AB - PURPOSE: To study cataract formation in eyes with an implantable contact lens (ICL) used for moderate to high myopia. SETTING: University Eye Hospital, Lausanne, Switzerland. METHODS: An ICL (model V3 or V4, Staar Surgical AG) was implanted in 75 eyes. Three months after surgery and again at the last follow-up examination, the transparency of the crystalline lens was assessed on transilluminated photographs and the vaulting of the ICL over the crystalline lens was evaluated. Central vaulting was measured precisely on digitized photographs taken with a 75 SL Zeiss slitlamp camera, while peripheral vaulting was estimated on photographs obtained with a Scheimpflug camera. The minimum follow-up was 12 months; the mean was 21.8 months. RESULTS: At the last follow up, 20 of the 75 eyes (27%) had an ICL-induced anterior subcapsular cataract (ASCC). The number of cataracts increased with the duration of the follow-up. Cataracts developed more commonly in older patients than in younger patients. All 20 cataracts occurred when the central vaulting was equal to or less than 0.09 mm. In 26 eyes with the same range of vaulting (among which 11 had no vaulting), the lenses were clear at the last visit. The 20 patients with cataract and the 26 patients with clear lenses matched in age and duration of follow-up but not in myopia. No touch between the ICL and the crystalline lens was encountered when the central vaulting was equal to or greater than 0.15 mm. Vaulting showed a slight decrease over time. No statistically significant difference in vaulting was found between models V3 and V4. CONCLUSION: Central and/or peripheral contact between the ICL and the crystalline lens may be responsible for the high incidence of ASCC formation in this study. Central vaulting greater than 0.09 mm appears to protect the crystalline lens from cataract formation. However, we recommend aiming for higher central vaulting (0.15 mm) to avoid contact between the ICL and the crystalline lens. This should be attainable by implanting longer ICLs. PMID- 12781277 TI - Pseudophakic retinal detachment after uneventful phacoemulsification and subsequent neodymium: YAG capsulotomy for capsule opacification. AB - PURPOSE: To determine the incidence of retinal detachment (RD) after neodymium:YAG (Nd:YAG) laser capsulotomy for posterior capsule opacification (PCO) and define the characteristics of this type of RD. SETTING: A private practice, with statistical evaluation at a university department. METHODS: Five hundred twenty-six consecutive eyes that had Nd:YAG capsulotomy because of visually disturbing PCO were followed prospectively for up to 52 months (median 21 months) to determine the incidence of RD. The median age of the 142 men and 320 women was 76 years and the median axial length, 23.3 mm (range 20.5 to 31.3 mm). Before the capsulotomy, all eyes had uneventful phacoemulsification with implantation of a posterior chamber intraocular lens (PC IOL). Eyes with additional surgical procedures or with panretinal laser photocoagulation were excluded. Eyes in which a retinal hole had been treated before Nd:YAG (n = 4) were included. The fundus was examined by binocular indirect ophthalmoscopy with a 2.2 diopter Volk lens in full mydriasis. If necessary, an examination with a 3 mirror lens was done. RESULTS: The incidence of RD was 0% (0/483; 95% confidence interval [CI], 0.0%-0.8%) at 6 months, 0% (0/407; 95% CI, 0.0%-0.9%) at 12 months, and 0.5% (1/213; 95% CI, 0.0%-2.6%) at 24 months. Twenty-one months after Nd:YAG capsulotomy, 1 eye developed an RD that occurred as the result of a horseshoe tear that reopened. The tear, which formed 7 months after phacoemulsification and PC IOL implantation and 6 years before Nd:YAG capsulotomy, had been immediately treated with argon laser photocoagulation. Excluding the 4 eyes with preexisting tears, presumably sealed retinal holes, the incidence at 24 months was 0% (0/212; 95% CI, 0.0%-1.7%). No other eye developed RD over the follow-up. CONCLUSIONS: Retinal detachment after Nd:YAG capsulotomy for PCO was rare in eyes that had previous uneventful phacoemulsification and PC IOL implantation. The exact relationship between the 2 events remains to be established. PMID- 12781278 TI - Posterior vitreous detachment after neodymium:YAG laser posterior capsulotomy. AB - PURPOSE: To determine whether retinal detachment (RD) after neodymium:YAG (Nd:YAG) laser posterior capsulotomy is due to a greater incidence of posterior vitreous detachment (PVD) than in controls and whether vitreous status at the time of capsulotomy is useful in predicting the risk for RD. SETTING: Teaching hospital ophthalmology service. METHODS: Patients having Nd:YAG laser posterior capsulotomy after uneventful cataract surgery (treatment group) were prospectively studied. Fellow eyes that had extracapsular cataract extraction (ECCE) with intraocular lens (IOL) implantation but no Nd:YAG capsulotomy (no laser group) formed 1 control group, and eyes that had no cataract surgery (phakic group) formed a second control group. The treatment group comprised 322 eyes; the no-laser group, 97; and the phakic group, 142. Dilated fundus and vitreous examinations were performed at baseline (before Nd:YAG capsulotomy) and 12 months postoperatively. RESULTS: At baseline, the prevalence of PVD was similar in the treatment and no-laser groups (61.8% and 63.9%, respectively; P=.2014) but was significantly lower in the phakic group (50.7%; P=.0151). There was no significant difference among the groups in the development of PVD in eyes with attached vitreous at baseline (17.9%, treatment group; 11.4%, no-laser group; 17.1%, phakic group) (P=.6588). CONCLUSIONS: The prevalence of PVD was significantly higher in eyes after ECCE and IOL implantation than in phakic eyes independent of Nd:YAG laser posterior capsulotomy. Capsulotomy was not associated with a significantly higher incidence of new PVD; therefore, the presence or absence of PVD at the time of capsulotomy is not helpful in assessing the risk for RD in the first year after laser treatment. PMID- 12781279 TI - Early rotational stability of the longer Staar toric intraocular lens: fifty consecutive cases. AB - PURPOSE: To determine whether rotational stability is improved with the longer Staar toric intraocular lens (IOL). SETTING: A private practice, Los Altos, California, USA. METHODS: Staar Surgical manufactures toric plate-haptic IOLs under +24.0 diopters (D) in 2 lengths. Fifty consecutive eyes requiring toric IOL spherical powers less than +24.0 D received the longer toric IOL (TL; 11.2 mm) using a specific surgical protocol. The IOL axis was recorded at 1 day, 1 week, and a final follow-up visit. Five additional eyes requiring powers greater than +24.0 D received the shorter toric IOL (TF; 10.8 mm), which is the only available length in these powers. Results were compared with those in an initial consecutive series of 6 eyes having implantation of the TF IOL with a power less than +24.0 D before the TL model became available. RESULTS: Rotational stability of all 50 TL IOLs and the 5 TF IOLs of higher dioptric powers was excellent. One IOL rotated as much as 20 degrees off axis. The repositioning rate was zero in both groups. In the initial 6 eyes receiving the shorter toric IOL in powers below +24.0 D, the repositioning rate was 50%. CONCLUSIONS: The longer toric IOL had excellent early rotational stability. Rotational and repositioning rates with the TL IOL were superior to those in a previous series of the TF IOL. Adequate overall length is a critical factor in the rotational stability of plate-haptic toric IOLs, and the longer IOL should be used when available. PMID- 12781280 TI - Impact of cataract surgery on self-reported visual difficulties: comparison with a no-surgery reference group. AB - PURPOSE: To examine the impact of cataract surgery on older adults' self-reported visual difficulties and compare them with those of patients with cataract who declined surgery over the same period. SETTING: Twelve area practices. METHODS: This was a consecutive chart review over a 6-month period. Primary inclusion criteria were 55 years or older, cataract in 1 or both eyes with 20/40 visual acuity or worse (best corrected, distance), and no previous cataract surgery in either eye. The Activities of Daily Vision Scale (ADVS) and visual acuity, contrast sensitivity, and disability glare tests were administered at baseline and at a 1-year follow-up visit. RESULTS: This study comprised 245 patients, 156 of whom elected to have cataract surgery and 89 of whom declined. Those electing surgery were more likely to be white, female, and have worse visual acuity and no ocular comorbidities. At baseline, ADVS subscale scores ranged from 53 to 76 in the surgery group and from 72 to 89 in the no-surgery group. In the surgery group, subscale scores improved by 15 to 21 points on average at the 1-year follow-up; scores were unchanged or worse in the no-surgery group over this period. This difference between the groups remained statistically significant after adjustment for group baseline differences in demographics, vision, and ADVS score. In the surgery group, visual acuity improvement in the first eye was an independent predictor of increases in the ADVS overall score and night driving and glare disability subscales; contrast sensitivity was an independent predictor of improvement in the night driving subscale. A reduction in disability glare in the second eye was independently linked to increases in the overall ADVS score and the night driving, near vision, and glare disability subscales. CONCLUSIONS: Baseline findings suggest that cataract patients who have surgery have more difficulty in visual tasks than those who decline surgery. After surgery, patients reported less difficulty with visual tasks. In the no-surgery group, no change was reported. Improvements in visual acuity and contrast sensitivity and reductions in disability glare after surgery were independently linked to improvements in ADVS scores. PMID- 12781282 TI - After-cataract in adults with primary posterior capsulorhexis: comparison of hydrogel and silicone intraocular lenses with round edges after 2 years. AB - PURPOSE: To evaluate the potential of primary posterior continuous curvilinear capsulorhexis (PCCC) to prevent after-cataract 2 years after surgery and perform a bilateral comparison of 2 intraocular lens (IOL) materials, silicone and hydrogel. SETTING: Department of Ophthalmology, University of Vienna, Medical School, Vienna, Austria. METHODS: In this randomized patient- and examiner-masked study, 37 patients (74 eyes) with age-related cataract had bilateral small incision cataract surgery with PCCC. Randomly, 1 eye received a hydrogel IOL and the contralateral eye, a silicone IOL. Both IOLs had open-loop haptics and a round-edged optic. Standardized digital retroillumination photographs were taken 1 day and 1, 6, 12, and 24 months after surgery to evaluate development of after cataract on the anterior and posterior capsules by subjective grading. Opacification of the PCCC area was also objectively evaluated using new software. RESULTS: Twenty-nine patients (58 eyes) completed the 2-year follow-up. Partial closure with ongrowth at the edge of the PCCC was found in 55% in the hydrogel group and 28% in the silicone group. Total closure of the PCCC was observed in 3 eyes, 2 in the hydrogel group and 1 in the silicone group. CONCLUSION: Ongrowth onto the PCCC area was dependent on IOL material. Because of less ongrowth and more fibrotic after-cataract with silicone IOLs, the efficacy of the PCCC was higher in this group. PMID- 12781281 TI - Suture fixation of a posterior chamber intraocular lens in anticoagulated patients. AB - PURPOSE: To describe a technique of suture fixating posterior chamber silicone intraocular lenses (PC IOLs) to the iris through a limbal incision and to evaluate its effectiveness in preventing intraocular hemorrhages. SETTING: Price Vision Group, Indianapolis, Indiana, USA. METHODS: This retrospective single center study comprised 7 eyes (5 patients) that had suture fixation of a silicone PC IOL from a limbal approach to the midperipheral iris using 9-0 polypropylene (Prolene(R)) suture. All patients were taking warfarin sodium (Coumadin(R)) preoperatively and postoperatively. At each follow-up examination, visual acuity and intraocular pressure were measured, the stability of the PC IOL was assessed, and the anterior chamber and posterior segment were evaluated for blood. RESULTS: There was no incidence of intraocular hemorrhage during surgery in any eye. In 6 eyes, there was no blood in the anterior chamber or posterior segment at any postoperative examination. A nonlayering hyphema that resolved within 1 week without sequelae was noted in 1 patient 6 weeks postoperatively. The mean follow up was 10.7 months (range 6 to 25 months). CONCLUSION: Suture fixating a PC IOL to the iris through a limbal approach was an effective means of placing an IOL in anticoagulated patients without capsule support. PMID- 12781283 TI - Ultrasound axial length measurement in biphakic eyes. AB - To determine the calculations to correct the error in ultrasound measurement of a phakic eye with a phakic intraocular lens (IOL) (biphakic eye) to increase the accuracy of IOL power calculation should cataract surgery become necessary. Mathematical derivations were performed to calculate the error caused by a phakic IOL while measuring an eye using an average sound velocity. A simple model was developed to allow correction of the axial length (AL). If the eye is measured at an average velocity of 1555 meters/second, the following calculation can be used depending on the material of the phakic IOL: AL(corrected) = AL(1555) + X x T, where T is the central thickness of the phakic IOL and X = +0.42 (0.41 - 0.42) for poly(methyl methacrylate), -0.59 (0.56 - 0.59) for silicone, +0.11 (0.10 - 0.12) for Collamer, and +0.23 (0.23 - 0.24) for acrylic. By obtaining the dioptric power of the phakic IOL and central thickness from the manufacturer and knowing the material of the lens, a mathematical correction can be made to arrive at the true ultrasound AL in a biphakic eye. PMID- 12781284 TI - Surgical management of postoperative endophthalmitis: comparison of 2 techniques. AB - PURPOSE: To evaluate the results of 2 surgical techniques in eyes with postoperative endophthalmitis. SETTING: Department of Ophthalmology, Dokuz Eylul University, School of Medicine, Izmir, Turkey. METHODS: Twenty-four eyes with endophthalmitis after cataract surgery had vitrectomy as an initial procedure according to the Endophthalmitis Vitrectomy Study (EVS) criteria (Group 1, n = 24). These eyes were compared with 28 eyes that had total pars plana vitrectomy with an encircling band, silicone tamponade, and endolaser (Group 2, n = 28). The visual and anatomical outcomes and the need for additional procedures (repeat vitrectomy) were evaluated in the 2 groups. RESULTS: In Group 1, 6 eyes (25.0%) had an additional procedure, 3 eyes (12.5%) had phthisis, and 21 eyes (87.5%) had successful surgery. In Group 2, no eye had an additional procedure, 1 eye (3.5%) had phthisis, and 27 eyes (96.4%) had successful surgery. The number of additional procedures was significantly less and the rate of surgical success was significantly higher in Group 2 than in Group 1 (P<.01). CONCLUSION: Despite the poor visual prognosis of endophthalmitis surgery, more radical intervention can increase the chance of surgical success and decrease the number of additional procedures in eyes with postoperative endophthalmitis. PMID- 12781285 TI - Evaluation of refractive error measurements of the Wavescan Wavefront system and the Tracey Wavefront aberrometer. AB - PURPOSE: To evaluate the accuracy and repeatability of the WaveScan WavePrint system and the Tracey wavefront aberrometer in measuring refractive errors in phakic eyes. SETTING: Cullen Eye Institute, Baylor College of Medicine, Houston, Texas, USA. METHODS: Using subjective manifest refraction (MR) as the standard, the spherical equivalent (SE), sphere, and cylinder were compared to values measured by WaveScan and Tracey devices in virgin eyes and eyes that had had corneal refractive surgery. Astigmatism was evaluated using vector analysis. The accuracy of the WaveScan and Tracey devices was assessed by 95% limits of agreement (95% LA), and repeatability was analyzed by 2 standard deviations (SDs) and intraclass correlation coefficients (ICCs). RESULTS: The mean differences in SE, sphere, and cylinder between MR and WaveScan were -0.26 diopter (D), -0.12 D, and -0.28 D, respectively, and between MR and Tracey, -0.21 D, -0.01 D, and -0.40 D, respectively. The 95% LA for SE, sphere, and cylinder were -1.09 to 0.57 D, 1.14 to 0.89 D, and -0.95 to 0.40 D, respectively, for WaveScan and -1.37 to 0.95 D, -1.27 to 1.26 D, and -1.16 to 0.35 D, respectively, for Tracey. Vector analysis revealed mean differences of -0.47 +0.07 x 9 degrees between MR and WaveScan and of -0.53 +0.27 x 12 between MR and Tracey. The 2 SDs for SE, sphere, and cylinder were 0.26 D, 0.29 D, and 0.16 D, respectively, for WaveScan and 0.31 D, 0.36 D, and 0.33 D, respectively, for Tracey. The ICCs for SE, sphere, and cylinder were 0.993, 0.992, and 0.902, respectively, for WaveScan and 0.994, 0.992, and 0.764, respectively, for Tracey. The Tracey device measured all eyes evaluated; the WaveScan could not measure 14% of normal eyes and 50% of post laser in situ keratomileusis eyes. CONCLUSIONS: Using MR as the standard, refractive errors measured by the WaveScan and Tracey devices were reliable and reproducible. However, the Tracey device was more robust in its ability to obtain measurements in normal and postoperative eyes. PMID- 12781286 TI - Unfolder lens injection system with acrylic intraocular lenses: retrospective study. AB - PURPOSE: To examine the efficacy of using the Unfolder Sapphire series system (Allergan) for implanting acrylic intraocular lenses (IOLs) with a 6.0 mm optic (Sensar AR40, Allergan). SETTING: Ambulatory day-surgery center. METHODS: Phacoemulsification was performed using topical anesthesia. The Unfolder injection system was used to insert a Sensar AR40 acrylic IOL. Data on complications and the success of implantation were collected retrospectively. RESULTS: Two hundred two consecutive patients having implantation of the Sensar IOL with the Unfolder system were examined. Three problems related to the Unfolder injector were identified. Another patient required an anterior vitrectomy, sulcus IOL implantation with the Unfolder, and optic capture; the complication was not related to the Unfolder system. Sulcus implantation was relatively easy with the Unfolder. CONCLUSION: Implantation of the Sensar AR40 IOL with the Unfolder system was easy to perform, and no serious complications occurred. PMID- 12781287 TI - Nuclear sclerotic cataract in young patients in Taiwan. AB - PURPOSE: To document the clinical features of discrete nuclear sclerosis in young patients. SETTING: Chang-Gung Memorial Hospital, Taoyuan, Taiwan, Republic of China. METHODS: The medical records of all patients younger than 45 years with nuclear sclerosis who had cataract surgery at 1 hospital were retrospectively reviewed. Patients with congenital cataract, ocular trauma, ocular disease in addition to myopia, and systemic disease associated with cataract were excluded. The patients' age, sex, bilateral lens status, axial length, original and preoperative refractive status, and preoperative and postoperative best corrected visual acuities (BCVAs) were recorded. RESULTS: Fifty-five eyes of 35 patients aged 32 to 45 years met the study criteria. Nuclear sclerotic cataract was bilateral in 20 patients. The mean age at surgery was 40.64 years +/- 3.63 (SD). The mean refraction before the development of cataract was -7.80 +/- 3.57 diopters (D) and the mean preoperative refraction, -17.40 +/- 6.05 D. The mean axial length was 28.09 +/- 2.31 mm. The postoperative BCVA was 20/22 or better in 53 eyes (96.4%). CONCLUSIONS; In patients younger than 45 years with cataract in Taiwan, nuclear sclerosis was not a rare cause of visual loss and axial myopia was strongly associated with the formation of nuclear sclerosis. Bilateral sclerosis was observed in most cases. Patients with an axial length greater than 27.0 mm had the greatest risk. Surgical treatment was therapeutic. PMID- 12781289 TI - Increased levels of advanced glycation end products in human cataractous lenses. AB - PURPOSE: To investigate the occurrence of advanced glycation end products (AGEs) formed oxidatively (pentosidine and N(epsilon)-carboxymethyl lysine [CML]) or nonoxidatively (imidazolone) in human lenses and the relation of AGEs to lens coloration, cataract type, and patients' diabetic state. SETTING: Departments of Ophthalmology and Internal Medicine III, University of Jena, Jena, Germany. METHODS: Pentosidine, CML, and imidazolone concentrations were measured in the water-soluble protein fraction of 44 cataractous lenses (from 24 nondiabetic and 20 diabetic donors) and 6 noncataractous control lenses. RESULTS: Pentosidine, CML, and imidazolone were higher in cataractous lenses than in control lenses (pentosidine, 3.7 pmol/mg +/- 5.3 (SD) and 1.9 +/- 1.7 pmol/mg, respectively; CML, 3.0 +/- 2.2 nmol/mg and 1.3 +/- 0.7 nmol/mg, respectively; imidazoline, 80.4 +/- 93.3 AU/mg and 19.6 +/- 18.5 AU/mg, respectively). Among the cataractous lenses, the highest AGE concentrations were found in mature cataracts, with a statistically significant increase in CML. The AGE content increased relative to the intensity of brown coloration of the lens; the brown coloration also indicated the highest rise of imidazolone compared to pentosidine and CML. Lenses from diabetic donors had generally similar pentosidine values and elevated CML and imidazolone levels compared to lenses from nondiabetic donors. The pentosidine, CML, and imidazolone levels in the lenses correlated significantly with one another but not with patient age. CONCLUSION: Advanced glycation end products formed oxidatively and nonoxidatively occurred to a higher degree in cataractous lenses than in noncataractous lenses. The strong relationship between the lenses' AGE content, color/opacity, and the state of the cataract may indicate that advanced glycation plays a pivotal role in cataract formation. PMID- 12781288 TI - Effects of topical nonsteroidal antiinflammatory drugs on the expression of matrix metalloproteinases in the cornea. AB - PURPOSE: To assess the effects of nonsteroidal antiinflammatory drug (NSAID) eyedrops on the expression of matrix metalloproteinases in corneal tissue. SETTING: Ocular Microbiology and Immunology Laboratory, Refractive Surgery Research Laboratory, The Wilmer Eye Institute, Johns Hopkins University, Baltimore, Maryland, USA. METHODS: Seventy rats were divided into 2 groups: intact and debrided epithelium. Uniform central corneal epithelial defects were created in the right eye of the debrided corneal group. Each group was divided into 4 subgroups, each receiving 1 of the following eyedrops or artificial tears: The 3 NSAIDs were diclofenac sodium 0.1% (Falcon or Voltaren) and preservative free ketorolac 0.5% (Acular PF). The artificial tears were carboxymethylcellulose sodium 0.5% (Refresh Plus PF). The eyedrops were administered 4 times a day for 1 week. The rats were killed on days 2 and 7. The corneas were excised and processed for immunohistochemical staining, Western blot assay, and zymography studies to determine the localization of the production of the following matrix metalloproteinases (MMPs): MMP-1, MMP-2, MMP-8, and MMP-9. RESULTS: Matrix metalloproteinase-1, MMP-8, and MMP-2 were detected in rat corneas at 48 hours in the debrided and intact epithelium groups treated with NSAID eyedrops. The MMP-1 and MMP-8 expression levels were higher in intact corneas in the diclofenac sodium groups than in the ketorolac and artificial tears groups. The expression was localized mostly in the epithelial cells and occasionally in keratocytes. CONCLUSION: This study provides preliminary evidence that topical application of some NSAIDs can induce the early expression of MMP-1, MMP-2, and MMP-8 in the cornea, suggesting that MMPs play a role in the corneal cytotoxicity of certain NSAIDs. PMID- 12781290 TI - Feasibility and safety of laser treatments in eyes with an intraocular implantable miniature telescope. AB - PURPOSE: To evaluate the feasibility and safety of laser treatments in rabbit eyes fitted with an implantable miniature telescope (IMT). SETTING: The Goldschleger Eye Research Institute, Tel-Aviv University, Sheba Medical Center, Tel-Hashomer, Israel. METHODS: In this study, IMTs were implanted in the capsular bag in 19 eyes of 19 pigmented rabbits. After the rabbits recovered from surgery, neodymium:YAG (Nd:YAG) laser iridectomy (4 eyes), Nd:YAG laser capsulotomy (8 eyes), or argon laser photocoagulation of the retina (6 eyes) was performed. One day after the laser treatment, the rabbits were killed. An observer blinded to the treatment received by individual rabbits examined the enucleated eyes histopathologically to evaluate the laser lesions and their effect on ocular tissues. RESULTS: Laser iridectomy and laser posterior capsulotomy were carried out without difficulty. Laser photocoagulation of the posterior pole and the retinal periphery could be accomplished only when the laser beam was aimed around the optic of the IMT. It was not possible to carry out effective laser treatment of the retina through the IMT's optic. Treatment was difficult when adhesions between the pupil margin and the IMT optic were encountered. CONCLUSIONS: Standard laser treatments in eyes with an IMT in the capsular bag were feasible in pigmented rabbits, whose eyes are a good model for the eyes of human patients. Laser treatment of the retina was difficult to perform when adhesions between the pupil margin and the IMT were present. PMID- 12781291 TI - Postoperative Morganella morganii endophthalmitis associated with subclinical urinary tract infection. AB - We report a case of Morganella morganii acute endophthalmitis following clear corneal phacoemulsification cataract surgery in which a coincident asymptomatic chronic urinary tract infection was detected postoperatively. Morganella morganii is a gram-negative bacillus that inhabits the gastrointestinal tract and is part of the normal fecal flora. It is an opportunistic pathogen usually encountered in postoperative and nosocomial settings, causing urinary tract and wound infections. Chronic urinary tract infection may be a risk factor for postoperative endophthalmitis. A dipstick urinalysis before elective cataract surgery in elderly patients with a history of recurrent urinary tract infections may be considered. PMID- 12781292 TI - Rapid diagnosis and treatment of mycobacterial keratitis after laser in situ keratomileusis. AB - We report the results of laser in situ keratomileusis (LASIK) in a 51-year-old woman with subsequent mycobacterial keratitis diagnosed by staining with acid fast and fluorochrome methods, a technique known to have good sensitivity and specificity for mycobacteria. A rapid diagnosis was made without waiting for cultures, and treatment was instituted, including tapering of topical steroids and appropriate antibiotic therapy. The result was preservation of the LASIK flap and a favorable visual outcome at 6 months. PMID- 12781293 TI - Curvularia keratitis after laser in situ keratomileusis from a feline source. AB - Fungal keratitis following laser in situ keratomileusis (LASIK) is extremely rare. Except in cases that follow trauma, the source of fungal organisms is unknown in published reports. We report a case of keratitis following LASIK that could be directly traced to a fungal skin infection of the patient's pet cat. PMID- 12781294 TI - Intraocular ointment after small-incision cataract surgery causing chronic uveitis and secondary glaucoma. AB - Anterior uveitis and secondary glaucoma resulting from intraocular ointment has not been reported. The advent of small-incision surgery has likely reduced the incidence of this complication to low levels. We report a case of anterior uveitis after small-incision cataract surgery due to an intraocular ointment base. The course of this rare case is described and the literature reviewed. PMID- 12781295 TI - Acute psychotic reaction caused by topical cyclopentolate use for cycloplegic refraction before refractive surgery: case report and review of the literature. AB - A 56-year-old woman was evaluated for the surgical correction of hyperopia (+3.0 diopters). Two drops of cyclopentolate 1% were instilled in both eyes for measurement of the cycloplegic refraction and wavefront analysis. Immediately after the second instillation, the patient reported drowsiness, dizziness, nausea, and fatigue. Ten minutes later, stimulatory central nervous system symptoms in the form of restlessness, cheerfulness, and a 20-minute-long roar of laughter were observed, interrupted by a new sedative phase. Basic medical and neurologic examinations were unremarkable except for gait ataxia. Four hours later, the examination was continued uneventfully. As surgical treatment of refractive errors and measurement of cycloplegic refraction using cyclopentolate become more frequent, ophthalmologists should be aware of this unusual acute event. PMID- 12781296 TI - Exodeviation following monocular myopic regression after laser in situ keratomileusis. AB - We report a 44-year-old woman with intermittent left exotropia of 35 prism diopters at distance who initially exhibited alignment of both eyes after bilateral laser in situ keratomileusis (LASIK). The exophoria was not preserved due to myopic regression in the dominant eye. An uneventful LASIK treatment was performed to correct -11.00 -0.50 x 130 in the right eye and -13.50 -1.50 x 145 in the left eye. The aim was to achieve emmetropia in both eyes. Although an examination revealed exophoria at near and distance during the 6 months following refractive surgery, the tropic aspect of the divergent deviation appeared in the right eye following the myopic regression. Laser in situ keratomileusis is an effective option to achieve binocular visual quality in myopic anisometropic patients. However, myopic regression after LASIK may disrupt the binocular visual quality. PMID- 12781297 TI - Retinal detachment and phthisis bulbi after implantation of an iris prosthetic system. AB - We report 2 eyes of 2 patients who developed persistent inflammation, hypotony, and total retinal detachment after secondary implantation of an iris prosthetic system (IPS) in association with cataract surgery. The first patient had an uneventful history of 16 years despite a retained intraocular glass foreign body and the second patient, of 38 years after sustaining penetrating corneoscleral injury with iris tissue loss. Both patients were thoroughly assessed clinically, and the enucleated globe in the first patient was examined by light microscopy. Both eyes had chronic inflammation with retrocorneal and cyclitic membrane formation around the artificial iris-lens diaphragm. These cases suggest that implantation of an IPS combined with cataract surgery can trigger decompensation of posttraumatic eyes that had been stable over a long period. PMID- 12781298 TI - Late anterior traumatic subluxation of a circular in-the-bag intraocular lens associated with lack of posterior capsule opacification. AB - We report a case of traumatic subluxation of a circular poly(methyl methacrylate) intraocular lens (IOL) 5 years after surgery. Slitlamp examination revealed the lack of epithelial cells or fibrosis in the intact capsule bag. Trauma was minor, and no other complication was found in the eye and orbit. The subluxated IOL was easily relocated in the bag with a hook, revealing the poor capsule sealing. Among the factors ensuring the fixation of IOLs placed in the bag after continuous curvilinear capsulotomy are sealing of the anterior and posterior capsules and proliferation of epithelial cells from the equator of the bag. PMID- 12781299 TI - Laser in situ keratomileusis as treatment for anisometropia after scleral buckling surgery. AB - Scleral buckling is an excellent procedure for retinal reattachment but can induce disabling refractive errors. Laser in situ keratomileusis (LASIK) has been proven effective for the reduction of refractive errors induced by ophthalmic surgery. We describe the case of a 53-year-old man who developed symptomatic anisometropia after placement of a scleral buckle for repair of a rhegmatogenous retinal detachment (RD). After the scleral buckling procedure, he retained excellent best corrected visual acuity but could not tolerate spectacle or contact lens correction. Thirty-four months after the scleral buckling procedure, LASIK was performed to correct myopic astigmatism with excellent refractive and functional results. This case demonstrates that LASIK may be safe and effective for the correction of refractive errors induced by RD repair. PMID- 12781300 TI - Maneuver to minimize zonular stress during removal of ophthalmic viscosurgical devices. PMID- 12781301 TI - Simple cataract checklist for residents and high-volume surgeons. PMID- 12781302 TI - Cilium as a foreign body in the meibomian gland opening. PMID- 12781303 TI - The role of illness severity and illness representations in adjusting to multiple sclerosis. AB - OBJECTIVE: Multiple sclerosis (MS) is an incurable, chronic and unpredictable disease of the central nervous system. The purpose of this study was to investigate whether MS patients' illness representations impact on their adjustment to this debilitating illness even when controlling for the severity of their condition. METHODS: One hundred and sixty-eight MS patients completed a questionnaire booklet comprised of the Illness Perceptions Questionnaire-Revised and a range of adjustment variables including the Sickness Impact Profile, the Fatigue Scale, the Hospital Anxiety and Depression Scale and the Rosenberg Self Esteem Scale. The severity of patients' MS was measured by the type of MS, length of illness, remission status and ambulatory ability. RESULTS: Hierarchical multiple regression analyses demonstrated that illness severity accounted for the majority of the variance in physical and role dysfunction, while patients' illness representations were the most significant predictors of levels of social dysfunction, fatigue, anxiety, depression and self-esteem. CONCLUSIONS: Patients' illness representations play a significant role in adjustment to MS. These results suggest that a psychological intervention, which addresses patients' illness representations, may assist in their adjustment to MS. PMID- 12781305 TI - Measuring fatigue severity in primary care patients. AB - OBJECTIVE: We developed a new instrument to measure fatigue that synthesized information from existing instruments. METHODS: 35 candidate items and 4 formats for a new fatigue scale were obtained from 15 previously developed instruments. A new scale was developed using factor analysis on a data set of 409 primary care patients and validated on a sample of 816 additional subjects. RESULTS: Different formats for obtaining information about a given fatigue item gave similar results. The new 11 item scale contained four subscales: cognitive, fatigue, energy and productivity. Correlations between the four subscales ranged from.49 to.66. Patients with a higher fatigue score were much more likely to have lower health status, greater depression and more somatic symptoms. CONCLUSION: This new instrument may be useful in primary care and epidemiological studies to screen and monitor patients for fatigue severity and type. PMID- 12781306 TI - Mutilation anxiety differs among females with fibromyalgia and functional dyspepsia and population controls. AB - BACKGROUND: Studies using self-report rating scales suggest a considerable overlap regarding symptom complaints in patients with fibromyalgia (FM) and functional dyspepsia (FD), while clinical assessments point to important psychological differences. PURPOSE: To test the hypothesis that measurement of psychological state by means of content analysis of speech will demonstrate differentiation between the two patient groups and between patients with these disorders and age-matched population-based random sample controls. METHOD: The Giessener Symptom Complaints Checklist assessed somatic complaints. The computerised Gottschalk-Gleser content analysis method assessed psychological state in 42 females with FM, 17 females with FD and 48 population-based, randomly selected control subjects. RESULTS: FM patients score higher on mutilation anxiety than FD and control subjects. FD patients had the highest score for death anxiety. Mutilation anxiety and low hope score identified FM patients (sensitivity 68%, specificity 81%, overall classification 75%), but only 19% of the variation in total somatic complaints could be predicted from these or other psychological state scores. In FD patients, however, death anxiety explained 59% of the variance in gastrointestinal complaints. INTERPRETATION: Psychological state was differentiated among the three groups. Mutilation anxiety may be a psychological marker of an underlying neurobiological vulnerability for FM or may represent a secondary long-term consequence of chronic illness. In FD, death anxiety is directly related to symptom complaints, suggesting a stronger etiological association between emotions and somatic complaints in this disorder. PMID- 12781307 TI - Cognitive and emotional characteristics of alexithymia: a review of neurobiological studies. AB - OBJECTIVE: To review neurobiological studies of alexithymia in order to achieve a better understanding of the relationship between alexithymia and psychosomatic diseases and psychiatric illnesses. METHODS: Neurobiological studies of alexithymia were reviewed with a special focus on how emotional and cognitive elements of alexithymia are reflected in earlier research. RESULTS: Studies that have correlated alexithymia to corpus callosum dysfunctioning have mainly found impairments in cognitive characteristics of alexithymia, whereas from studies of right hemisphere and frontal lobe deficits, it may be concluded that both cognitive and emotional characteristics of alexithymia are impaired. CONCLUSION: The fact that there is no general agreement on how to define alexithymia seems to have hampered theoretical and empirical progress on the neurobiology of alexithymia and related psychosomatic diseases and psychiatric illnesses. Alexithymia should no longer be approached as one distinct categorical phenomenon and follow-up studies should monitor subjects according to both the cognitive and emotional characteristics of alexithymia. PMID- 12781308 TI - Are dysfunctional beliefs about illness unique to hypochondriasis? AB - OBJECTIVE: There is evidence that individuals high in hypochondriasis overestimate the likelihood of ambiguous symptoms being indicative of serious illness. However, it is not known whether this tendency is unique to hypochondriasis or whether it can be attributed to high negative affectivity or other anxiety symptoms often found to be comorbid with hypochondriasis. METHOD: College students (N=133) completed measures of hypochondriasis, depression, anxiety, worry, avoidance and estimated the likelihood of various symptoms indicating catastrophic and minor illnesses. RESULTS: Even after entering the other self-report variables, hypochondriasis was the only variable to predict estimates of the likelihood of serious illness. Conversely, being female, high levels of negative affect, agoraphobic avoidance when accompanied by others and higher estimates about the likelihood of symptoms leading to catastrophic illnesses best predicted hypochondriasis scores. CONCLUSION: Dysfunctional beliefs about illness appear to be unique to hypochondriasis and to uniquely contribute to the prediction of hypochondriasis. PMID- 12781309 TI - Severity of allergic complaints: the importance of depressed mood. AB - OBJECTIVE: To evaluate the prevalence of depressive symptoms in patients with different kinds of allergic diseases and the connection of depressive symptoms with the severity, type and seasonality of allergic complaints. METHODS: Data was obtained via a cross-sectional multicenter questionnaire survey of 528 patients aged 16-60 years attending six regional in- and outpatient allergy clinics in Hungary in June to July 1998. Consecutive patients completed a structured, self administered questionnaire containing questions about their current and past allergic complaints. Depressive symptoms were measured by the Shortened Beck Depression Inventory (BDI). RESULTS: 32.2% of patients scored above the normal level (> or =10) and 12.5% had clinically significant depressive symptomatology (> or =19) by the BDI. These rates were significantly higher than those found in the control group from a national representative population sample (22.4% and 8.3%). Patients with asthma and with perennial symptoms had significantly higher depression scores than patients with other types of allergic diseases. There was a significant association between the severity of depressive symptoms and the severity of allergic complaints independently from age, sex, type and seasonality of the allergic disease, and other current physical illnesses and symptoms tested by the General Linear Model (GLM). CONCLUSIONS: Our results draw attention that patients even with mild depressive symptoms have significantly more severe allergic complaints and assess general health state as much worse than those without depressive symptoms in any types of allergic diseases. Diagnosis and treatment of depressive symptoms in allergic patients is of great concern from both a clinical and an economical point of view. PMID- 12781310 TI - Mood, eating attitudes, and anger in obese women with and without Binge Eating Disorder. AB - OBJECTIVE: The aim of this study was to evaluate the anger levels and their management in obese patients. METHODS: A total of 103 obese women [51 with Binge Eating Disorder (BED) and 52 without BED] were included in the study and compared to 93 healthy controls. They were assessed with the State-Trait Anger Expression Inventory (STAXI), Beck Depression Inventory (BDI), and Eating Disorder Inventory 2 (EDI-2). RESULTS: The BDI score is higher in obese subjects than in controls and obese binge eaters have higher levels of depression than obese patients without BED. Differences among the three groups can be found in almost all subscales of the EDI-2, even after controlling for the variable depression (BDI). For STAXI, the only difference among the three groups, which remains significant after controlling for depression, is the tendency to express anger outside (AX OUT), which is higher in obese binge eaters. The correlation study highlights the importance of impulsivity in the group of obese binge eaters, whereas in obese patients without BED, the tendency toward anger suppression (AX-IN) is seen. DISCUSSION: Obese patients with BED might be considered a subgroup deserving greater psychiatric interest, both for the greater severity of the eating disorder and for the comorbidity with subthreshold depressive symptoms and with borderline personality traits. In obese patients without BED, eating behavior seems more correlated to the psychological functioning typical of psychosomatic disorders. Implications for treatment are discussed. PMID- 12781311 TI - The use of the Pros and Cons of Anorexia Nervosa (P-CAN) Scale with children and adolescents. AB - OBJECTIVE: The Pros and Cons of Anorexia Nervosa (P-CAN) Scale was developed out of a previous qualitative study [Int J Eat Disord 25 (1999) 177] in which the positive and negative aspects of anorexia nervosa (AN) were elicited from individuals with AN. Principal components analysis was then used in an adult sample to reduce the number of items and to derive empirically and theoretically based subscales [Serpell et al., submitted]. The aim of the current study was to validate the P-CAN in children and adolescents with AN and compare the pros and cons of young people with AN with those of adults. METHODS: Forty-eight young people with AN completed the P-CAN and the Eating Disorders Examination, a semistructured interview used to aid in the diagnosis of eating disorders. RESULTS: Validity in younger patients appeared to be good. Children and adolescents showed both similarities and differences from adults tested in a previous study. CONCLUSIONS: The measure is the first to measure attitudes of children and adolescents with AN towards their illness and may prove a useful tool for the measurement of attitudes towards AN in both clinical and research settings. PMID- 12781312 TI - Anxiety sensitivity and 35% CO2 reactivity in patients with panic disorder. AB - OBJECTIVE: The present study examines the possible relationships between anxiety sensitivity (AS) and reactivity to the 35% carbon dioxide (CO(2)) challenge in panic disorder (PD). METHODS: One-hundred eight patients with PD underwent the 35% CO(2) challenge and completed the Anxiety Sensitivity Index (ASI). Multiple regression analyses were applied to evaluate the role of AS as a predictor of CO(2)-induced anxiety. RESULTS: Fifty-six patients with PD showed high AS scores, whereas 48 showed medium scores and 4 low scores. ASI scores significantly predicted symptomatological reaction to CO(2) but not subjective induced anxiety. CONCLUSION: These findings suggest that the fear of anxiety-related bodily sensations was related to the symptomatological reactivity to CO(2) but did not seem to play a crucial role in the modulation of the subjective anxiogenic/panicogenic response to hypercapnia in patients with PD. PMID- 12781313 TI - Effect of changing referral mode to C-L Psychiatry for noncognitively impaired medical inpatients with emotional disorders. AB - OBJECTIVE: To test the effect of changing referral mode to Consultation-Liaison Psychiatry (C-L Psychiatry) by implementing early screening with systematic multidisciplinary management for emotional disorders in noncognitively impaired medical inpatients. METHOD: A prospective pre- and postintervention controlled study in the internal medicine division of a university hospital. RESULTS: Out of 515 patients admitted to the internal medicine ward during the study period, 176 were included in the study and gave their informed consent (male=63%); 81 in the control group and 95 in the intervention group. Except for the increase in referral rate to the C-L Psychiatry service (4-32%), there were no significant differences between the baseline and intervention periods either in terms of length of stay in the internal medicine ward and of use and costs of medical resources, or in terms of patients' satisfaction as regards attention paid to psychosocial issues during hospitalization. CONCLUSION: As a result of changing referral mode to C-L Psychiatry, the lack of effect on length of stay and on medical consumption of medically ill inpatients should be considered in the context of the briefness of the hospitalization periods encountered. Furthermore, the change in referral process does not seem to increase patients' perception of the quality of care provided. PMID- 12781314 TI - Treatment efficacy in behavioral pediatric sleep medicine. AB - Behavioral interventions have been identified as the treatment of choice for many forms of pediatric sleep disturbance. We adopt criteria established by the Task Force on Promotion and Dissemination of Psychological Procedures (1996) to evaluate the literature base describing behavioral interventions for pediatric sleep disturbance. Three well-established interventions, one promising intervention and one probably efficacious intervention, have emerged for bedtime refusal and frequent nighttime awakenings. Only one intervention for sleep terrors and sleepwalking has sufficient empirical support to be considered promising. Behavioral interventions targeting circadian rhythm disorders (e.g., delayed sleep phase), nightmares and rhythmic movement disorder (RMD) have not been researched sufficiently to be considered empirically supported. PMID- 12781315 TI - Nasal CPAP treatment for obstructive sleep apnea: developing a new perspective on dosing strategies and compliance. AB - Nasal continuous positive airway pressure (CPAP) is presently considered as the "treatment of choice" for obstructive sleep apnea (OSA). Though some OSA patients adhere to treatment recommendations and ultimately respond quite well to CPAP therapy, there is a substantial subgroup for which compliance is a particularly difficult issue. Despite receiving recommendations to the contrary and for reasons that are incompletely understood, the majority of OSA patients settle for a partial compliance pattern. Whether a partial compliance schedule is beneficial or harmful is virtually unexamined: Unlike other medical treatments, there are few data concerning the "dose-response relationship" of CPAP to its desired effects. We argue that CPAP "dose" is a function not only of CPAP pressure but of time-on-CPAP as well. Critical questions that remain unanswered are what "dose" of CPAP is needed to effect an appropriate treatment outcome and which treatment outcomes should form the basis of our recommendations. Recent placebo-controlled studies comparing CPAP to suboptimal CPAP pressures may be informative in this regard. Directions for future research are suggested. PMID- 12781321 TI - Experimentally-induced microencephaly: effects on cortical neurons. AB - Genetic and epigenetic factors may alter the normal development of cerebral cortex, producing laminar and cellular abnormalities and heterotopiae, major causes of juvenile, drug-resistant epilepsy. Experimentally-induced migration disorders provide interesting insights in the mechanisms of the determination of neuronal phenotype and connectivity, of congenital cortical dysgenesis and the pathophysiology of associated neurological disorders, such as epilepsy. We investigated the effects of E14 administration of methylazoxymethanol acetate (MAM), which induces microencephaly by ablating dividing cells. Brains from newborn and adult rats were reacted for NADPH-d and CO histochemistry. Moreover, callosally-projecting neurons were retrogradely labeled with DiI at P9 or with BDA in adults. MAM-treated rats displayed a remarkable reduction in cortical thickness, mainly due to reduction in layer IV and in supragranular layers. Heterotopic nodules appeared in the supragranular layers and in the hippocampus. CO-positive barrels in somatosensory cortex were almost absent. The distribution of NADPH-d-positive neurons was regular, but they were rare in heterotopic nodules. Callosally-projecting neurons displayed abnormal orientation of the apical dendrite and increase in the basal dendritic length. Alterations in the dendritic arborization of pyramidal neurons may be one of the substrates for the increased sensitivity to drugs which induce epileptic seizures in these animals. PMID- 12781320 TI - Role of citron kinase in dendritic morphogenesis of cortical neurons. AB - Small GTPases of the rho family regulate the extensive rearrangements of the cytoskeleton that characterize neuronal differentiation. Citron kinase is a target molecule for activated rhoA, previously implicated in control of cytokinesis. We have found that, in addition, it could play an important role in modulating the extension of neuronal processes. Using constitutively active and dominant negative mutants, we showed that citron kinase is involved in the morphologic differentiation of N1E-115 neuroblastoma cells induced by serum starvation. More importantly, quantitative analysis of citron kinase knockout cerebral cortex displayed that this molecule may differentially regulate the morphology of the dendritic compartment in corticocollicular versus callosally projecting pyramidal neurons. PMID- 12781322 TI - Organization of cortico-cortical associative projections in rats exposed to ethanol during early postnatal life. AB - The fine organization of cortico-cortical associative projections was investigated in adult rats exposed to inhalation of ethanol during the first postnatal week. Ethanol-treated and control animals received cortical injections of biotinylated dextran amine combined with N-methyl-D-aspartic acid, in order to obtain a Golgi-like retrograde labeling of associative pyramidal neurons. The results obtained from the analysis of labeling can be summarized as follows: (a) there are fewer associative projection neurons in ethanol-treated than in normal animals; (b) the ratio between the number of supragranular and infragranular associative neurons is higher in ethanol-treated animals compared to controls; (c) the basal dendrites of pyramidal associative cells of layer 2/3 display a simplified dendritic branching in ethanol exposed cases as compared to controls; (d) the cluster analysis shows that normal dendrites can be clearly subdivided into different groups according to their geometric properties, whereas dendrites from animals exposed to ethanol follow less robust grouping criteria. These differences are discussed in consideration of the functional alterations that characterize the fetal alcohol syndrome. PMID- 12781323 TI - Cellular physiology of the neonatal rat cerebral cortex. AB - The early development of the cerebral cortex is characterized by neurogenesis, neuronal migration, cellular differentiation and programmed cell death. Cajal Retzius cells, developing cortical plate neurons and subplate cells form a transient synaptic circuit which may serve as a template for the formation of cortical layers and columns. These three neuronal cell types show distinct electrophysiological properties and synaptic inputs. Endogenous or exogenous harmful disturbances during this developmental period may lead to the preservation of early cortical circuits, which may act as trigger zones for the initiation of pathophysiological activity. PMID- 12781324 TI - Development of functional thalamocortical synapses studied with current source density analysis in whole forebrain slices in the rat. AB - We analysed the laminar distribution of transmembrane currents from embryonic (E) day 17 until adulthood after selective thalamic stimulation in slices of rat forebrain to study the development of functional thalamocortical and cortico cortical connections. At E18 to birth a short-latency current sink was observed in the subplate and layer 6, which was decreased, but not fully abolished in a cobalt containing solution or after the application of glutamate receptor blockers (APV and DNQX). This indicated that embryonic thalamic axons were capable of conducting action potentials to the cortex and some of them had already formed functional synapses there. Between birth and P3, when thalamic axons were completing their upward growth, a sink gradually appeared more superficially in the dense cortical plate and synchronously, a current source aroused in layer 5. Both sinks and sources completely disappeared after blocking synaptic transmission. The adult-like distribution of CSDs became apparent after P7. The component in layer 6 cannot be blocked completely after this age suggesting antidromic activation. This study demonstrated that cells of the lowest layers of the cortex received functional thalamic input before birth and that thalamocortical axons formed synapses with more superficial cells as they grew into the cortical plate. PMID- 12781325 TI - Effect of developmental sensory and motor deprivation on the functional organization of adult rat somatosensory cortex. AB - Most sensory systems are active, in the sense that the animal performs specific motor actions in order to collect information of interest-signals are not merely passively received. We, therefore, expect cortical development to depend not only correct sensory experience, but also on correct motor experience. In this study, we used the rat whisker system as a model to compare the importance of these factors. In one group of animals, we trimmed all whiskers starting from post natal day 8 (P8). In a second group, we left the whiskers intact, but prevented "whisking" by sectioning the facial (VIIth cranial) nerve on P8. The first group had severely disrupted sensory experience but normal motor patterns ("whisker cut" rats); the second group had normal sensory pathways within which temporal activity patterns were disrupted by motor impairment ("nerve-cut" rats). When they reached 3 months of age, we recorded multi-unit responses from the infragranular layers of primary somatosensory cortex in response to deflection of either single whiskers or pairs of whiskers in order to compare these two groups to a third group of rats that had normal sensory and motor experience. Cortical topographic organization was unaltered in whisker- and nerve-cut rats. Whisker cut rats showed a smaller than normal difference between the response magnitudes for the principal and surrounding whiskers, as well as stronger than normal interactions between co-active whisker inputs. Responses in nerve-cut rats were nearly indistinguishable from those in normal animals. Thus, unexpectedly, neither pure sensory nor sensorimotor deprivation caused gross functional disruption of SI according to our measures. It appears that abnormal sensory experience leads to alterations in the fine-tuning of cortical properties, but cortex is unexpectedly resistant to the effects of abnormal sensory and sensorimotor experience. PMID- 12781326 TI - The evolution of mammalian cortex, from lamination to arealization. AB - We analyse some of the most important anatomical and functional features emerging at different stages of mammalian brain evolution in terms of a possible computational advantage. At the transition from reptiles to mammals, a major anatomical change occurs in the originally sensory dorsal cortex. The principal layer of pyramidal cells is split by the insertion of a new layer of granule cells, giving rise to the laminated isocortex. It has been hypothesized that this qualitative change in the evolution of mammalian brains is necessary to support fine topography in their sensory maps. The simulation of neural network models demonstrates that a nonlaminated patch of cortex must compromise between transmitting "where" information, explicitly mapped, topographically, on the cortical sheet, and retrieving "what" information, represented by the distributed firing pattern across neurons. The differentiation of a granular layer is shown in the model to yield a small quantitative advantage, allowing to transmit a slightly better combination of both information types. Along the same theoretical lines, we are investigating the multiplication of successive sensory areas coding for ever more composite stimuli, such as those in the visual and auditory temporal cortices in primates. In particular we analyse the possible computational advantage for a specific neural population devoted to encode the complex structure of whole stimuli, rather than relying on the coactivation of separate populations encoding their basic elements. PMID- 12781327 TI - Opioid-induced immunosuppression: is it centrally mediated or peripherally mediated? AB - Opioid compounds are commonly used pain medications. However, their administration is associated with a number of side-effects. Among them, opioid induced immunosuppression is a significant medical problem, which is evidenced by a strong association between the use of opioids and exacerbated infections, including AIDS. Research data have demonstrated the effects of opioids to be suppressive on phagocytic, natural killer (NK), B and T cells. However, these immunosuppressive effects may be mediated by mechanisms different from those for antinociceptive actions. This article reviews possible central and peripheral mechanisms of opioid-induced immunosuppression. To the extent that peripherally mediated immunosuppressive effects play a significant role in opioid-induced immunosuppression, novel peripheral opioid antagonists may have a therapeutic role in attenuating opioid-induced immunosuppression without affecting analgesia. PMID- 12781328 TI - Non-caspase-mediated apoptosis contributes to the potent cytotoxicity of the enediyne antibiotic lidamycin toward human tumor cells. AB - Enediyne antibiotics have been reported to be the most potent cytotoxic antitumor agents. The pathway by which these compounds cleave DNA and induce apoptosis of tumor cells may be different from the caspase-mediated pathways that initiate typical apoptosis. In this report, we studied the apoptosis induced by lidamycin (LDM), a member of the enediyne antibiotic family, and compared the characteristics of LDM-induced apoptosis with those of typical apoptosis induced by mitomycin C or etoposide. Chromatin condensation occurred very rapidly and appeared as speckles in human hepatoma BEL-7402 and breast carcinoma MCF-7 cells after treatment with 1 microM LDM. In addition, co-staining the cells with the mitochondria-specific dye Mitosensor and the DNA-specific dye Hoechst 33342 enabled the visualization of mitochondria in normal control and LDM-treated cells but not in mitomycin C-treated cells. Neither the caspase inhibitor VAD-fmk nor the caspase-3 inhibitor DEVD-fmk was able to inhibit the DNA ladder patterns caused by LDM in BEL-7042 or MCF-7 cells. Smaller fragments of histone H1 cleaved by LDM were detected by SDS-PAGE, indicating that the site of LDM action is the internucleosomal structure. Although caspase-9, caspase-3/7, and caspase-6 activities were increased in BEL-7402 cells, and caspase-7 activity was increased in MCF-7 cells after treatment with 1 microM LDM, this occurred much later, indicating that chromatin condensation reached the maximal level rapidly while caspase activities still remained low. Taken together, these results demonstrate that LDM induced rapid DNA cleavage and chromatin condensation independently of caspase activities; this may contribute to its highly potent cytotoxicity toward tumor cells. PMID- 12781329 TI - Induction of G1 phase arrest in MCF human breast cancer cells by pentagalloylglucose through the down-regulation of CDK4 and CDK2 activities and up-regulation of the CDK inhibitors p27(Kip) and p21(Cip). AB - Pentagalloylglucose (5GG) is a potent and specific inhibitor of NADPH dehydrogenase or xanthine oxidase. In our previous study, we showed that 5GG was able to induce apoptosis in HL-60 cells in a time- and concentration-dependent manner via the activation of caspase-3. Recently, we found that 5GG was capable of perturbing the cell cycle of the human breast cancer cell line MCF-7. DNA flow cytometric analysis showed that 5GG exhibited the ability of blocking MCF-7 cell cycle progression at the G1 phase. The level of several G1 phase-related cyclins and cyclin-dependent kinases did not change in these cells during a 24-hr exposure to 5GG. However, the activity of cyclin E/CDK2 was decreased in a concentration- and time-dependent manner and the activity of cyclin D/CDK4 was inhibited when serum-starved synchronized cells were released from synchronization. p27(Kip) and p21(Cip), inhibitors of cyclin/CDK complexes in G1 phase, were gradually increased after 5GG treatment in a time-dependent manner and the induction of p21(Cip) was correlated with an increase in p53 levels. These results suggest that the suppression of cell-cycle progression in the G1 phase by 5GG was mediated in MCF-7 cells, at least in part, by either the inhibition of cyclin D/CDK4 and cyclin E/CDK2 activity or the induction of the CDK inhibitors p27(Kip) and p21(Cip). PMID- 12781331 TI - Selection of insulin-producing rat insulinoma (RINm) cells with improved resistance to oxidative stress. AB - The defense system against reactive oxygen species is believed to be crucial for the survival of insulin-producing cells after various injuries. The aim of our study was to select a subpopulation of insulin-producing RINm cells with higher resistance to oxidative stress. The cells resistant to hydrogen peroxide (RINmHP) were obtained by repeated exposure of parental RINm cells to 100 and 200 microM hydrogen peroxide (HP). The increased resistance of RINmHP cells to HP was confirmed by three different cytotoxicity assays. In addition, the selected cells also were resistant to the cytotoxic effect of activated rat splenocytes compared to parental cells. The half-life of HP in the RINmHP cell culture medium was about 2.5 times lower than that of the parental cells, corresponding to the increased level of catalase expression and activity in selected cells. The increased defense property of the selected cells was not associated with any significant changes in insulin content and insulin response to a mixture of glucose with isobutyl methyl xanthine or potassium chloride. In conclusion, repeated exposure to HP induces selection of RINm cells with improved resistance to oxidative stress. This improved defense characteristic probably is due to an increased level of catalase expression and activity in the selected cells. PMID- 12781330 TI - Pharmacogenetics of human 3'-phosphoadenosine 5'-phosphosulfate synthetase 1 (PAPSS1): gene resequencing, sequence variation, and functional genomics. AB - 3'-Phosphoadenosine 5'-phosphosulfate (PAPS) is the high-energy "sulfate donor" for reactions catalyzed by sulfotransferase (SULT) enzymes. The strict requirement of SULTs for PAPS suggests that PAPS synthesis might influence the rate of sulfate conjugation. In humans, PAPS is synthesized from ATP and SO(4)(2 ) by two isoforms of PAPS synthetase (PAPSS): PAPSS1 and PAPSS2. As a step toward pharmacogenetic studies, we have resequenced the entire coding sequence of the human PAPSS1 gene, including exon-intron splice junctions, using DNA samples from 60 Caucasian-American and 58 African-American subjects. Twenty-one genetic polymorphisms were observed-1 insertion-deletion event and 20 single nucleotide polymorphisms (SNPs)-including two non-synonymous coding SNPs (cSNPs) that altered the following amino acids: Arg333Cys and Glu531Gln. Twelve pairs of these polymorphisms were tightly linked, and a total of twelve unequivocal haplotypes could be identified-two that were common to both ethnic groups and ten that were ethnic-specific. The Arg333Cys polymorphism, with an allele frequency of 2.5%, was observed only in DNA samples from Caucasian subjects. The Glu531Gln polymorphism was rare, with only a single copy of that allele in a DNA sample from an African-American subject. Transient expression in mammalian cells showed that neither of the non-synonymous cSNPs resulted in a change in the basal level of enzyme activity measured under optimal assay conditions. However, the Glu531Gln polymorphism altered the substrate kinetic properties of the enzyme. The Gln531 variant allozyme had a 5-fold higher K(m) value for SO(4)(2-) than did the wild-type allozyme and displayed monophasic kinetics for Na(2)SO(4). The wild type allozyme (Glu531) showed biphasic kinetics for that substrate. These observations represent a step toward testing the hypothesis that genetic variation in PAPS synthesis catalyzed by PAPSS1 might alter in vivo sulfate conjugation. PMID- 12781332 TI - Improved potency of the hypoxic cytotoxin tirapazamine by DNA-targeting. AB - To improve the potency of the hypoxic cytotoxin tirapazamine (TPZ), we have constructed an analog, SN26955, with the TPZ moiety attached to an acridine chromophore to target the drug to DNA. The underlying reason for this is our previous finding that the hypoxic cytotoxicity of TPZ is a result of its ability to produce DNA double-strand breaks, whereas many of the toxicities of the drug in clinical use are likely the result of its metabolism in the cytoplasm and effects on mitochondria. We found that the DNA-targeted TPZ analog was more potent than TPZ in killing hypoxic cells by 1-2 orders of magnitude, yet it retained the hypoxic selectivity for cell killing of TPZ. We show that SN26955 is only active in producing DNA damage when it is enzymatically reduced while bound to, or in close association with, the DNA. We also show that it has a different cofactor dependence than TPZ for reduction leading to DNA double-strand breaks, suggesting the involvement of a different reductase for production of the lethal lesion than for TPZ. These results show the promise of DNA-targeting of TPZ to produce a DNA compound with greater clinical efficacy than TPZ itself. PMID- 12781333 TI - An amperometric biosensor with human CYP3A4 as a novel drug screening tool. AB - We developed a biosensor based on the redox properties of human CYP3A4 to directly monitor electron transfer to the heme protein. Enzyme films were assembled on gold electrodes by alternate adsorption of a CYP3A4 layer on top of a polycation layer. Direct, reversible electron transfer between the electrode and CYP3A4 was observed with voltammetry under anaerobic conditions. In the presence of oxygen, the oxidation peak of the hemoprotein disappeared, and the reduction peak increased 2- to 3-fold. Addition of CYP3A4 substrates (verapamil, midazolam, quinidine, and progesterone) to the oxygenated solution caused a concentration-dependent increase in the reduction current in cyclic voltammetric and amperometric experiments. Product analyses after electrolysis with the enzyme film showed catalytic activity of the biosensor depending on substrate concentration, its inhibition by ketoconazole, and a minor contribution of H(2)O(2) to the catalytic cycle. These results suggest that electron exchange between the electrode and the immobilized CYP3A4 occurred, and that metabolic activity of the enzyme was maintained. Thus, important requirements for the application of human CYP biosensors in order to identify drugs or drug candidates as substrates or inhibitors to the attached enzyme are fulfilled. PMID- 12781334 TI - Large-conductance K+ channel openers NS1619 and NS004 as inhibitors of mitochondrial function in glioma cells. AB - Recently, it has been reported that large-conductance Ca(2+)-activated potassium channels, also known as BK(Ca)-type potassium channels, are present in the inner mitochondrial membrane of the human glioma LN229 cell line. Hence, in the present study, we have investigated whether BK(Ca)-channel openers (BK(Ca)COs), such as the benzimidazolone derivatives NS004 (5-trifluoromethyl-1-(5-chloro-2 hydroxyphenyl)-1,3-dihydro-2H-benzimidazole-2-one) and NS1619 (1,3-dihydro-1-[2 hydroxy-5-(trifluoromethyl)phenyl]-5-(trifluoromethyl)-2H-benzimidazol-2-one), affect the functioning of LN229 glioma cell mitochondria in situ. We examined the effect of BK(Ca)COs on mitochondrial membrane potential, mitochondrial respiration and plasma membrane potassium current in human glioma cell line LN229. We found that BK(Ca)COs decrease the mitochondrial membrane potential with an EC(50) value of 3.6+/-0.4 microM for NS1619 and 5.4+/-0.8 microM for NS004. This mitochondrial depolarization was accompanied by an inhibition of the mitochondrial respiratory chain. Both BK(Ca)COs induced whole-cell potassium current blocked by charybdotoxin, as measured by the patch-clamp technique. The BK(Ca)COs had no effect on membrane bilayer conductance. Moreover, the inhibition of mitochondrial function by NS004 and NS1619 was without effect on cell survival, as measured by lactate dehydrogenase release from the cells. PMID- 12781335 TI - K-ATP channel independent effects of pinacidil on ATP production in isolated cardiomyocyte or pancreatic beta-cell mitochondria. AB - Evidence has been presented that mitochondria contain ATP sensitive potassium channels (mK-ATP channels), which may confer tissue protection upon activation. It is, however, not known whether activation of mK-ATP channels has a direct effect on mitochondrial ATP production. This study was performed to define the effect of pinacidil (PIN) on ATP production by oxidative phosphorylation in isolated cardiomyocyte or pancreatic beta-cell mitochondria. Cardiomyocyte mitochondria produced seven times more ATP than beta-cell mitochondria in the presence of pyruvate/malate. PIN inhibited pyruvate/malate-induced mitochondrial ATP production with half maximal effect at 360 microM in both cell types. The inclusion of 5-hydroxydecanoate (5-HD) did not prevent this inhibition. Succinate induced a similar ATP production in cardiomyocyte or beta-cell mitochondria. In beta-cell mitochondria succinate-induced ATP production was inhibited by PIN with half maximal effects at 500 microM PIN. However, in cardiomyocyte mitochondria PIN stimulated succinate-induced ATP production 3-fold with half maximal effect at 100 microM and maximal effect at 200 microM. This PIN-dependent stimulation was mimicked by rotenone. The inclusion of 5-HD could not prevent these PIN effects. In conclusion, PIN may inhibit complex 1 of the respiratory chain without indications of opening mK-ATP channels. In cardiomyocytes with metabolically inhibited succinate dehydrogenase this results in a stimulation of ATP production conferring tissue protection. In beta-cells without a metabolically inhibited succinate dehydrogenase, there is no stimulation by PIN and tissue protection by PIN is not to be expected. PMID- 12781336 TI - Human MDR1 polymorphism: G2677T/A and C3435T have no effect on MDR1 transport activities. AB - The two most frequently observed single nucleotide polymorphisms (SNPs) of the human multidrug resistance 1 (MDR1) gene are 2677G/T/A (893Ala/Ser/Thr) and 3435C/T (no amino acid substitution). In this study, six forms of MDR1 cDNAs with the SNPs were expressed in LLC-PK1 cells and their transport activities were determined. Nearly identical amounts of the recombinant MDR1 proteins were expressed in the established cell lines using the Flp recombinase, which integrates a gene of interest at a specific genomic location. Four structurally diverse compounds: verapamil, digoxin, vinblastine and cyclosporin A, were examined for transcellular transport activities and intracellular accumulation. No significant differences were observed between cells expressing five polymorphic types of the MDR1 cDNAs (2677G/3435T, 2677A/3435C, 2677A/3435T, 2677T/3435C, 2677T/3435T) and cells expressing the wild-type (2677G/3435C). These results suggested that the two frequently observed MDR1 SNPs had no effect on the transport activities of MDR1 proteins expressed in LLC-PK1 cells in vitro, and other genetic or environmental factors might control the expression of MDR1 and the in vivo activity of MDR1. PMID- 12781337 TI - Preclinical factors affecting the interindividual variability in the clearance of the investigational anti-cancer drug 5,6-dimethylxanthenone-4-acetic acid. AB - Cancer chemotherapy is characterized by significant interindividual variations in systemic clearance, therapeutic response, and toxicity. These variations are due mainly to genetic factors, leading to alterations in drug metabolism and/or target proteins. The aim of this study was to determine, using a human liver bank (N=14), the interindividual variations in the expression and activity of liver enzymes that metabolize the investigational anticancer drug 5,6 dimethylxanthenone-4-acetic acid (DMXAA), i.e cytochrome P450 (CYP1A2) and uridine diphosphate glucuronosyltransferase (UGT1A9/2B7). In addition, interindividual variations in enzyme inhibition, hydrolysis of DMXAA acyl glucuronide (DMXAA-G) by plasma and hepatic microsomes, and the binding of DMXAA by plasma proteins also were examined. The results indicated that there was approximately one order of magnitude of interindividual variation in the expression of CYP1A2 and UGT2B7, activity of the enzymes toward DMXAA, and inhibition potency (IC(50)) by diclofenac, cyproheptadine, and alpha naphthoflavone. The enzyme activities toward DMXAA and IC(50) values were closely correlated with enzyme expression. There was a smaller (2- to 3-fold) variation in the enzyme-catalyzed hydrolysis of DMXAA acyl glucuronide in human plasma and liver microsomes (N=6) and in the binding of DMXAA by plasma proteins in humans. In conclusion, the interindividual variability of DMXAA disposition observed in vitro might reflect the greater elimination variability (>one order of magnitude) in Phase I cancer patients. The variability in DMXAA clearance in these cancer patients would be due mainly to differences in its metabolism and its metabolic inhibition by co-administered drugs. To a lesser extent, variability in the clearance of DMXAA could be due to the hydrolysis of its acyl glucuronide and/or its binding to plasma proteins. Further study is needed to examine the genotype phenotype relationship, and the result, together with therapeutic drug monitoring may provide a useful strategy for optimizing DMXAA treatment. PMID- 12781338 TI - Monoamine oxidase A inhibitory potency and flavin perturbation are influenced by different aspects of pirlindole inhibitor structure. AB - Reversible inhibitors of monoamine oxidase A (MAO A) are used as antidepressants. The influence of inhibitors such as pirlindole (pyrazinocarbazole) on the redox co-factor (flavin adenine dinucleotide, FAD) is a key factor in the inhibition. The kinetic, spectral, and thermodynamic changes induced by four closely related pirlindole analogues have been determined to investigate their interaction with the FAD in the active site of MAO A. For a model of flavin-inhibitor stacking, more favourable association would be expected between lumiflavin and the flatter analogues with a double bond at N3, and indeed lower K(i) values were found. However, the spectral changes induced by inhibitor binding to MAO A were 45% less for inhibitors with a double bond. Both in the absence and presence of the double bond, compounds with cyclohexyl at C8 induced 85% larger decrease in absorbance at 500nm than did those with a methyl substituent. In contrast, the K(i) values for the cyclohexyl compounds were lower, indicating greater affinity despite the lower perturbation of the flavin spectrum. All inhibitors stabilised the semiquinone of the FAD when MAO A was titrated with dithionite and prevented further reduction. These results indicate that the active site of MAO A is far more sensitive to structural variation than would be predicted by the simple flavin stacking model. Further, the independent changes in inhibitory potency and flavin perturbation preclude direct interaction with the flavin as a mode of binding and indicate that inhibitor-protein interactions must be important for inhibition. PMID- 12781339 TI - Cytosolic beta-glycosidases for activation of glycoside prodrugs of daunorubicin. AB - Human cytosolic beta-glycosidase is a small monomeric enzyme that is active under physiological conditions, which might be ideal for enzyme-prodrug therapy. We have previously reported the synthesis of a galactoside (DNR-GlA3) and a glucoside (DNR-GsA3) prodrug of daunorubicin. In the present study, we established that cellular uptake of DNR-GlA3 and DNR-GsA3 was low in contrast to that of daunorubicin. Recombinant human beta-glycosidase converted both prodrugs to daunorubicin as shown by liquid chromatography. The kinetics of the conversion of DNR-GlA3 and DNR-GsA3 by human beta-glycosidase, however, was unfavorable as the K(m) values were, respectively, 3- and 6-fold higher than those of another mammalian beta-glycosidase of bovine origin. The V(max) values were, respectively, 3.3 and 8.5nmol/hr/mg as compared to 158.3 and 147.8nmol/hr/mg of the bovine enzyme. Treatment of OVCAR-3 cells with human beta-glycosidase (0.5U/mL) and 0.5 microM DNR-GlA3 or DNR-GsA3 resulted in, respectively, 86 and 81% cell growth inhibition, while the prodrugs alone inhibited growth to only 19 and 1%. Treatment of cells with the bovine enzyme and the prodrugs inhibited cell growth more efficiently. We conclude that the endogenous intracellular beta glycosidase is not available for extracellular prodrug activation. Thus, the incorporation of the enzyme in enzyme-prodrug therapy might be an elegant approach to achieve tumor-specific prodrug conversion. The efficiency of glycoside prodrug conversion might be improved by design of a prodrug that is more readily activated by human beta-glycosidase or by evolution of the enzyme into a mutant form that displays high activity towards these prodrugs. PMID- 12781340 TI - Inhibition of leptin release by atrial natriuretic peptide (ANP) in human adipocytes. AB - The addition of atrial natriuretic peptide (ANP) to isolated human adipocytes in primary culture from very obese individuals resulted in an inhibition of leptin release after a 24- or 48-hr incubation. There was also an inhibition of leptin release by isoproterenol (ISO) that was partially reversed by insulin, whereas the inhibition due to ANP was unaffected. Similar results were seen with N-[2-(p bromocinnamylamino)ethyl]-5-isoquinolinesulphonamide (H-89), which is a cell permeable inhibitor of protein kinase A. H-89 markedly reduced the effects of ISO on both lipolysis and leptin release without affecting the stimulation of lipolysis or the inhibition of leptin release due to ANP. Inhibition of endogenous nitric oxide formation using N(omega)-nitro-L-arginine resulted in a 20% increase in leptin release over 48 hr, which suggests that the nitric oxide/cyclic GMP pathway might play a small role in the regulation of endogenous leptin release. Similarly, the addition of the nitric oxide donor (Z)-1-[2 aminoethyl)-N-(2-aminoethyl)diazen-1-ium-1,2-diolate (DETA NONOate) at 0.1 or 1 microM to explants of human adipose tissue enhanced lipolysis by 29%. Our data demonstrate that the lipolytic effect of ANP is probably secondary to stimulation of cyclic GMP accumulation in human adipocytes, and this is accompanied by an inhibition of leptin release. PMID- 12781341 TI - In vivo activation of the human CYP3A4 promoter in mouse liver and regulation by pregnane X receptors. AB - Human cytochrome P450 3A4 (CYP3A4) is responsible for the metabolism of numerous xenobiotics in the human liver. We have examined the activation of the human CYP3A4 promoter in mouse liver by using in vivo bioluminescent imaging (BLI). Transcription of the CYP3A4 promoter occurs as a result of a ligand binding to a nuclear orphan receptor, pregnane X receptor (PXR), followed by dimerization with another nuclear receptor, retinoid X receptor (RXR). Since this heterodimer then binds to xenobiotic response elements to activate transcription of CYP3A4, we examined a 13kb promoter region of CYP3A4 for responsiveness to dexamethasone and rifampicin. A reporter vector CYP3A4-luc was constructed consisting of the CYP3A4 promoter driving the firefly luciferase gene. This DNA was injected into the tail veins of mice, and reporter gene expression was monitored in the liver region using BLI. Treatment of transfected mice with dexamethasone resulted in a 188 fold induction of luciferase, whereas treatment with rifampicin resulted in a 68 fold induction. Co-injection with a human PXR expression vector resulted in a dramatic increase in rifampicin-induced activity and a smaller increase of dexamethasone-induced activity. Co-injection of an antisense murine PXR construct with the CYP3A4-luc reduced both the dexamethasone- and rifampicin-induced responses, thus demonstrating that the murine PXR receptor can participate in the regulation of the human CYP3A4 promoter in mice. The approach described here will be of general use in studying the regulation of nuclear receptors in vivo. PMID- 12781342 TI - Esculetin inhibits Ras-mediated cell proliferation and attenuates vascular restenosis following angioplasty in rats. AB - The proliferation of vascular smooth muscle cells (VSMCs) induced by injury to the intima of arteries is an important etiologic factor in vascular proliferative disorders such as atherosclerosis and restenosis. Esculetin, derived from the Chinese herb Artemisia scoparia, is well known as a lipoxygenase inhibitor. We have investigated the inhibitory effects of esculetin on VSMC proliferation and intimal hyperplasia by balloon angioplasty in the rat. We determined, using [3H]thymidine incorporation and the 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) assay, that esculetin inhibited the proliferation of VSMCs via a lipoxygenase-independent pathway. Three predominant signaling pathways were identified to be inhibited by esculetin: (a) the activation of p42/44 mitogen-activated protein kinase (MAPK) and the downstream effectors of c-fos and c-jun immediate early genes by means of western and reverse transcription-polymerase chain reaction (RT-PCR) analyses; (b) the activation of nuclear factor-kappaB (NF-kappaB) and activator protein-1 (AP-1), using the electrophoretic mobility shift assay; and (c) the activation of phosphoinositide 3-kinase (PI 3-kinase) and cell cycle progression, by western blot analysis and flow cytometric detection. Furthermore, esculetin also profoundly inhibited Ras activation, a shared upstream event of the above signaling cascades. In vascular injury studies, intraperitoneal administration of esculetin significantly suppressed intimal hyperplasia induced by balloon angioplasty. We conclude that esculetin blocks cell proliferation via the inhibition of an upstream effector of Ras and downstream events including p42/44 MAPK activation, PI 3-kinase activation, immediate early gene expression, as well as NF-kappaB and AP-1 activation. It also inhibits intimal hyperplasia after balloon vascular injury in the rat, indicating the therapeutic potential for treating restenosis after arterial injury. PMID- 12781343 TI - Intracellular signal transduction modulating expression of plasminogen activator inhibitor-1 in adipocytes. AB - The concentrations in blood of plasminogen activator inhibitor-1 (PAI-1), an inhibitor of fibrinolysis and proteolysis, are elevated in obese and insulin resistant subjects, predispose them to the risk of thrombosis, and may accelerate atherogenesis. Adipose tissue is a prominent source. Accordingly, intracellular signaling pathways that may influence PAI-1 expression in adipocytes have been the focus of considerable study. Rho, a small GTP binding and GTPase protein, when activated in turn activates its target, Rho-associated coiled-coil forming protein, to yield an active kinase, Rho-kinase, an effector in the Rho pathway. Rho-kinase exerts calcium-sensitizing effects in vascular smooth muscle cells and inhibitory effects on transforming growth factor-beta (TGF-beta) expression in chicken embryonic heart cells. Because TGF-beta is a powerful agonist of PAI-1 expression, we characterized the effects of inhibition of Rho-kinase in 3T3-L1 adipocytes. PAI-1 mRNA was determined by Northern blotting, and PAI-1 protein was determined by Western blotting. The Rho-kinase inhibitor, Y-27632 [(R)-(+)-trans N-(4-pyridyl)-4-(1-aminoethyl)-cyclohexanecarboxamide], increased PAI-1 expression markedly. Although genistein, a flavonoid tyrosine kinase, attenuated the increase of PAI-1 induced by Y-27632, other non-flavonoid tyrosine kinase inhibitors did not. However, another flavonoid, daidzein, which lacks tyrosine kinase activity, decreased basal PAI-1 expression and attenuated the induction of PAI-1 expression by Y-27632. Thus, the Rho/Rho-kinase system inhibits PAI-1 expression by a flavonoid-sensitive mechanism in adipocytes. Therefore, flavonoids may be useful in decreasing elevated PAI-1 expression in adipose tissue and its consequent pathophysiologic sequelae. PMID- 12781344 TI - Cyclothymic OCD: a distinct form? AB - BACKGROUND: Clinical research on the comorbidity of obsessive compulsive disorder (OCD) and other anxiety disorders has largely focused on depression. However in practice, resistant or severe OCD patients not infrequently suffer from a masked or hidden comorbid bipolar disorder. METHOD: The rate of bipolar comorbidity in OCD was systematically explored among 453 members of the French Association of patients suffering from OCD (AFTOC) as well as a psychiatric sample of OCD out patients (n=175). As previous research by us has shown the epidemiologic and clinical sample to be similar, we combined them in the present analyses (n=628). To assess mood disorder comorbidity, we used structured self-rated questionnaires for major depression, hypomania and mania (DSM-IV criteria), self-rated Angst's checklist of Hypomania and that for the Cyclothymic Temperament (French version developed by Akiskal and Hantouche). RESULTS: According to DSM-IV definitions of hypomania/mania, 11% of the total combined sample was classified as bipolar (3% BP-I and 8% BP-II). When dimensionally rated, 30% obtained a cut-off score >/=10 on the Hypomania checklist and 50% were classified as cyclothymic. Comparative analyses were conducted between OCD with (n=302) versus without cyclothymia (n=272). In contrast to non-cyclothymics, the cyclothymic OCD patients were characterized by more severe OCD syndromes (higher frequencies of aggressive, impulsive, religious and sexual obsessions, compulsions of control, hoarding, repetition); more episodic course; greater rates of manic/hypomanic and major depressive episodes (with higher intensity and recurrence) associated with higher rates of suicide attempts and psychiatric admissions; and finally, a less favorable response to anti-OCD antidepressants and elevated rate of mood switching with aggressive behavior. LIMITATION: Hypomania and cyclothymia were not confirmed by diagnostic interview by a clinician. CONCLUSION: Our data extend previous research on "OCD-bipolar comorbidity" as a highly prevalent and largely under-recognized and untreated class of OCD patients. Furthermore, our data suggest that "cyclothymic OCD" could represent a distinct form of OCD. More attention should be paid to it in research and clinical practice. PMID- 12781345 TI - Could mood state affect risk-taking decisions? AB - BACKGROUND: Depressed patients are advised by clinicians to avoid making critical life decisions because their decision-making process might be influenced by their pathological mood states. In order to provide empirical support for this practice, we examined the influence of different mood states on risk-taking tendencies that govern one's decision during critical life decisions. METHOD: By means of viewing a happy, neutral or sad movie clip, participants were induced to a respective mood. Risk-taking tendencies were then measured with decision tasks modified from the Choice Dilemmas Questionnaire (Kogan N., Wallach, M.A., 1964. Risk Taking: A Study in Cognition and Personality, Holt, New York). RESULTS: The findings (SS=54) indicated varying risk-taking tendencies in different mood states: individuals in induced depress mood were significantly more conservative in taking risk than those who were in neutral mood, whilst people in induced elated mood did not differ significantly from those in neutral mood. Correlation between mood-ratings with risk-taking tendency was positive (r=0.319, P<0.05). CONCLUSIONS: People in induced depress mood would have a lower willingness to take risk than people in neutral and in positive mood. The manner in which decisions were influenced by negative mood was discussed. The asymmetry of influence of different mood states on risk-taking decisions was also discussed. PMID- 12781346 TI - Perception of dyadic relationship and emotional states in patients with affective disorder. AB - BACKGROUND: Previous research has shown that interpersonal processes play a significant role in the development and maintenance of affective disorders. In this study, this claim was further investigated by comparing the perception of the dyadic relationship and judgment of other's emotions in affective disorder patients. METHOD: The sample included 39 couples (n=39 couples) with one of the partners suffering from an affective disorder and currently either in an acute or remitted depressive state. All participants completed four instruments, measuring the perceived quality of the dyadic relationship and the perception of other's emotions as reflected by judgments of facial expressions line drawings. RESULTS: While the level of marital satisfaction was found to be lower in the acute than in the remitted group both for ill partners and their spouses, spouses in both the acute and remitted group tended to be more critical of their ill partners. Patients who were depressed judged facial expressions significantly less positively than did remitted patients. Judgments of negative emotions were highly correlated between partners in the acute group, but uncorrelated in the remitted group. Acutely depressed patients were less sensitive to invitation than remitted patients, while their spouses displayed the opposite pattern. CONCLUSION: The present results shed further light on the interpersonal dynamics between depressed patients and their spouses by underscoring differences between couples with a remitted vs. acutely depressed partner in their perception of the dyadic unit and their judgments of facial emotions. LIMITATIONS: Longitudinal research is needed, in which the same patients are tested during periods of remission and acute episodes, as well as research investigating the role of patient gender in the perception of facial expressions of emotions. PMID- 12781347 TI - Factors associated with patients' knowledge of and attitudes towards treatment with lithium. AB - BACKGROUND: Treatment with lithium is often compromised by poor adherence, by side-effects and by patients' having serum levels outside the therapeutic range. These factors may be affected by patients' knowledge and attitudes towards lithium, and we set out to establish factors associated with knowledge about and attitudes towards lithium among a large representative sample of patients. METHOD: Patients known to be taking lithium in Grampian during 1995 were surveyed postally during 1998 with the Lithium Knowledge Test (LKT) and the Lithium Attitudes Questionnaire (LAQ). Scores on these measures were analysed against patients' sociodemographic and clinical characteristics by stepwise multiple regression. RESULTS: Of 742 patients, 411 (55%) completed an LKT and 362 (49%) completed an LAQ. Stepwise multiple regression established that positive attitudes towards lithium on the LAQ were associated with higher serum lithium levels (P=0.005) and with continuing to take lithium (P<0.001). Higher knowledge on the LKT was associated with positive attitudes on the LAQ (P=0.002), with younger age (P<0.001), and with shorter duration of treatment (P=0.01) LIMITATIONS: The study was retrospective and the response rate was relatively low. CONCLUSIONS: Education about lithium is likely to be of particular importance in the elderly and 'refresher courses' are advisable for those who have been on lithium for lengthy periods. Interventions which modify attitudes, rather than enhancing knowledge, are likely to be helpful in promoting adherence. PMID- 12781348 TI - Personality traits and personality disorders in early onset versus late onset major depression. AB - BACKGROUND: We aimed to determine the relationship between certain personality disorders and/or personality traits and early onset major depression. METHODS: A total of 400 depressed primary care patients were assessed for personality disorders using the SCID screen and for personality traits using the Karolinska Scales of Personality (KSP) questionnaire. Early onset was defined as onset of the first episode before the age of 26. Logistic regressions were performed to reveal relationships after adjustment for sex, age and number of previous episodes. RESULTS: Both groups had a similar severity of current illness determined by the Montgomery-Asberg Depression Rating Scale. Those with an early onset presented with a more debilitating course, seen in the form of more depressive episodes and previous hospitalisations in spite of their younger age. Early onset was also an independent predictor for avoidant, borderline and paranoid personality disorders. It also predicted increased scores on the KSP scales Psychic anxiety, Psychasthenia, Muscular tension, Suspicion and Irritability, and decreased Socialisation. LIMITATIONS: The evaluation was performed as a self-assessment, subjects had a superimposed major depressive episode when assessed, and subgroups of individuals were not eligible. CONCLUSIONS: Early onset major depression is a predictor for personality pathology and deviant personality traits. A better understanding of the interplay between genetics and environment that underlies this phenomenon will help to improve the long-term course in afflicted individuals. PMID- 12781349 TI - Factors associated with being granted a pension among psychiatric outpatients with major depression. AB - BACKGROUND: Little is known about factors associated with early retirement due to major depression in naturalistic settings. We examined to what extent major depression leads to disability pension and whether there are any associated factors with being granted a pension. METHODS: In our retrospective document based cohort study of 213 adult psychiatric outpatients with first-time documented DSM-III-R major depression, several sociodemographic, clinical and treatment characteristics were detected during the follow-up time of 3 months of medical care in Finland. This information was related to official registers of granted pensions with a follow-up time of 30 months. RESULTS: Forty-six (21.6%) patients were granted a pension during the follow-up period. Greater age, comorbidity and lowered self-esteem were strongly associated with being granted a pension. DISCUSSION: Some risk factors associated with subsequent retirement could be identified at the early phase of the illness. PMID- 12781350 TI - Consequences of displaying abnormal social behaviour: avoidance and reduction of social reinforcement. AB - BACKGROUND: Abnormal social behaviour, which is a common feature of psychiatric disorders, is associated with rejection. A passive lack of participation or involvement has been studied as characteristic of depression but active forms of nonparticipation have received little experimental attention. This study examined the interpersonal consequences of four distinct types of social behaviour by using the role enactment method. Two of the roles portrayed abnormal social behaviour, active nonparticipant 'manic' and passive nonparticipant 'sad', and two portrayed normal social behaviour, active participant 'warm' and passive participant 'shy'. METHODS: Sixty-three normal subjects were randomly allocated to a brief dyadic social interaction with a confederate acting one of four roles. Subsequently, they rated their level of rejection of the confederate and took part in the mixed-motive game with him/her. RESULTS: The subjects were more likely to reject confederates in the abnormal social behaviour roles. This was shown on both their nonverbal behaviour and their verbal report. On the mixed motive game, subjects gave fewer points and less cooperative and ingratiating messages to the confederates who had displayed abnormal social behaviour. LIMITATIONS: This result might only reveal the effects of first impressions of a confederate who behaves in a particular way, but not be generalised to long term acquaintanceship. CONCLUSIONS: These results extend previous findings that passive nonparticipant behaviour leads to rejection to active nonparticipant behaviour and show that the consequences of displaying such behaviour not only result in rejection but also in the reduction of social reinforcement. This might slow a patient's recovery process. PMID- 12781351 TI - Glia as a putative target for antidepressant treatments. AB - BACKGROUND: Since the early 1950s, various molecular mechanisms have been invoked to explain how antidepressants work. The most recent suggests that pharmacologically stimulated adult neurogenesis might be involved. Surprisingly, in the adult brain, an important source of new neurons and possibly mediators of neurogenesis appears to be glia, i.e., astrocytes. We have recently shown that protracted administration of the antidepressant fluoxetine to adult rats upregulated the astrocytic protein S100beta content and increased neurogenesis in the hippocampus. METHODS: Rats were treated with fluoxetine for 21 days; before sacrifice bromodeoxyuridine (BrdU) was injected to label the proliferating cells. Immunofluorescence was used to identify proliferating BrdU-positive cells, and cells immunopositive for S100beta and its receptor receptor for advanced glycation end products (RAGE). RESULTS: Typically, S100beta-positive cells were observed in the vicinity of BrdU-positive cells. On the other hand, we observed colocalization of RAGE receptors and BrdU immunoreactivities, suggesting that some proliferating cells express these receptors for S100beta. RAGE expression by neuronal cells or neuronal precursors and its activation by S100beta may promote their survival. LIMITATIONS: The anatomical localization of hippocampal S100beta, its receptor RAGE, and BrdU-positive cells that we describe in this study is only indicative of a putative role for glia in antidepressant-stimulated neurogenesis. Functional in vitro and in vivo studies are needed to directly investigate this role; quantitative assays and time-course studies are also warranted. CONCLUSION: We propose that a better understanding of glia functioning could establish its role as a target for novel antidepressant treatments. PMID- 12781352 TI - Studies on serotonergic markers in the human hippocampus: changes in subjects with bipolar disorder. AB - BACKGROUND: Various studies suggest the hippocampus and serotonergic systems are important in the pathology of bipolar disorder (BD). We therefore measured hippocampal serotonergic markers in post-mortem tissue from BD and control subjects. METHODS: The density and affinity of [3H]citalopram binding to the serotonin transporter (SERT), as well as the density of the 5HT(2A), 5HT(1A), 5HT(1D) and 5HT(1F) receptors were measured. RESULTS: The density of SERT and 5HT receptors was no different in BD. There was a significant decrease in the affinity of [3H]citalopram binding to SERT in the stratum lacunosum-moleculare (S(lac)) in BD (K(d) mean+/-S.E.M.=4.3+/-0.8 vs. 1.9+/-0.3 nM). LIMITATIONS: This study was completed using relatively small cohorts. CONCLUSIONS: There are no generalised changes in hippocampal serotonergic markers in the hippocampus from subjects with BD. There is a decreased affinity of radioligand binding to S(lac) SERT in subjects with BD. PMID- 12781353 TI - Validation of the Edinburgh Postnatal Depression Scale (EPDS) in Spanish mothers. AB - BACKGROUND: The EPDS is a 10-items self-report scale designed as a specific instrument to detect postnatal depression. It was validated in the UK and in other countries, but not in Spain. METHOD: We report data from all women (n=1201) attending in a routine postnatal check-up at 6 weeks postpartum during 1 year. A two-stage method was used: for the first stage, all individuals selected completed the EPDS; in the second one, 'probable cases' (n=261) with EPDS score >/=9, and a randomised sample of 10% with EPDS score less than 9 (n=126) were interviewed by a psychiatrist using the Structured Clinical Interview for DSM-IV, non-patient (SCID-NP), to establish psychiatric diagnosis of major and minor depression. RESULTS: The best cut-off of the Spanish validation of the EPDS was 10/11 for combined major and minor depression, the sensitivity was 79%, and specificity was 95.5%, with a positive predictive value of 63.2% and a negative predictive value of 97.7%. At this cut-off all cases of major depression were detected. The area under the ROC curve was 0.976 (P<0.0001) with an asymptotic interval of confidence of 95% between 0.968 and 0.984. CONCLUSIONS: Our data confirm the validity of the EPDS to identify postnatal depression in its Spanish version. The cut-off 10/11 for major and minor combined depression is useful to screen for a posterior psychiatric evaluation in Spanish sample. PMID- 12781354 TI - Altered dopamine D2 receptor function in fibromyalgia patients: a neuroendocrine study with buspirone in women with fibromyalgia compared to female population based controls. AB - BACKGROUND: To what extent fibromyalgia belongs to affective spectrum disorders or anxiety spectrum disorders remains disputed. Buspirone induces a hypothermic response, which most likely is due to 5-HT(1A) autoreceptor stimulation, and growth hormone (GH) release, which probably is related to postsynaptic 5-HT(1A) receptor stimulation. The prolactin response to buspirone has been suggested to be mediated through dopamine (DA) antagonistic effects. OBJECTIVES: Based on the assumption that fibromyalgia is more strongly related to stress and anxiety than affective spectrum disorders, we hypothesized that compared to population controls, fibromyalgia patients should demonstrate an increased prolactin response (DA sensitivity) to buspirone challenge test, but no difference in hypothermic response or GH release (5HT sensitivity). METHOD: A 60-mg dose of buspirone was given orally to 22 premenopausal women with fibromyalgia and 14 age and sex matched healthy control subjects. Core body temperature, growth hormone and prolactin levels were analyzed at baseline and after 60, 90, and 150 min. RESULTS: Fibromyalgia patients showed an augmented prolactin response to buspirone compared to controls. Temperature and growth hormone responses did not differ from controls. CONCLUSIONS: Dopaminergic rather than serotonergic neurotransmission is altered in fibromyalgia, suggesting increased sensitivity or density of dopamine D(2) receptors in fibromyalgia patients. Stress and anxiety is an important modulator of dopaminergic neurotransmission. Our results suggest that fibromyalgia is related to anxiety and associated with disturbance in the stress response systems. PMID- 12781355 TI - The clinical use of gabapentin in bipolar spectrum disorders. AB - BACKGROUND: with increasing awareness of lithium's limitations, several new anticonvulsants had been tested for their mood stabilisation during recent years. Among the innovative third generation mood stabilizing anticonvulsants, gabapentin (GBP) seems to have a broad spectrum of efficacy, although no certain data are available as to its efficacy and use in clinical practice. Accordingly, an extensive review on this subject has been carried out. METHODS: A computer generated search of the biomedical literature and abstract books of the more important scientific psychiatric congresses until June 2000 was undertaken to identify all pertinent case reports, case series and studies of GBP as monotherapy or adjunctive therapy in mood disorders. We identified 40 open-label studies on the use of GBP in at least 600 patients with bipolar disorder (BP), manic, depressed, or mixed episodes and unipolar depression and four controlled studies. RESULTS: The 40 open-label studies and two of the controlled trials suggested that GBP may have a role as adjunctive agent in the treatment of patients with bipolar disorders particularly when complicated by co-morbid anxiety disorder or substance abuse. GBP is usually very well tolerated and has no pharmacological interference with other mood stabilisers. However, in the other two double-blind studies GBP has not been found to be efficacious in the treatment of refractory mania or refractory bipolar depression. CONCLUSIONS: Although failing to show clear antimanic efficacy in randomized trials, gabapentin still remains a clinically useful agent when it comes to combination treatment in refractory and co-morbid patients. PMID- 12781356 TI - Transgenics at breaking-point. AB - In this issue of Cancer Cell, Forster et al. (2003) have generated mice that recapitulate both the mechanism (sporadic somatic translocation) and the consequences (expression of two translocation fusion genes) leading to an accurate leukemia model. PMID- 12781357 TI - Targeting oncogene dependence and resistance. AB - Our expanding experience with imatinib mesylate provides instructive lessons on the power and pitfalls of targeted therapy. The often impressive initial clinical responses seen with imatinib in a variety of malignancies inevitably give way to the emergence of resistant disease. Recent findings reveal several mechanisms of resistance and suggest ways to overcome them. PMID- 12781358 TI - FANCF methylation contributes to chemoselectivity in ovarian cancer. AB - A new model of ovarian cancer tumor progression implicates aberrant FANCF promoter methylation that is associated with gene silencing and disruption of the Fanconi-anemia-BRCA pathway. Disruption of the pathway occurs de novo in ovarian cancers and may contribute to selective sensitivity to platinum salts. PMID- 12781359 TI - Chk1 and Chk2 kinases in checkpoint control and cancer. AB - Accumulation of mutations and chromosomal aberrations is one of the hallmarks of cancer cells. This enhanced genetic instability is fueled by defects in the genome maintenance mechanisms including DNA repair and cell cycle checkpoint pathways. Here, we discuss the emerging roles of the mammalian Chk1 and Chk2 kinases as key signal transducers within the complex network of genome integrity checkpoints, as candidate tumor suppressors disrupted in sporadic as well as some hereditary malignancies and as potential targets of new anticancer therapies. PMID- 12781360 TI - Adoptive immunotherapy: engineering T cell responses as biologic weapons for tumor mass destruction. AB - Adoptive T cell immunotherapy is an evolving technology with the potential of providing a means to safely and effectively target tumor cells for destruction. PMID- 12781361 TI - PDGF receptors as cancer drug targets. PMID- 12781362 TI - Cell fusion: a hidden enemy? PMID- 12781363 TI - Engineering de novo reciprocal chromosomal translocations associated with Mll to replicate primary events of human cancer. AB - The etiology of human tumors often involves chromosomal translocations. Models that emulate translocations are essential to understanding the determinants of frank malignancy, those dictating the restriction of translocations to specific lineages, and as a basis for development of rational therapeutic methods. We demonstrate that developmentally regulated Cre-loxP-mediated interchromosomal recombination between the Mll gene, whose human counterpart is involved in a spectrum of leukemias, and the Enl gene creates reciprocal chromosomal translocations that cause myeloid tumors. There is a rapid onset and high penetrance of leukemogenesis in these translocator mice, and high proportions of cells carrying chromosomal translocations can be found in bone marrow as early as 12 days after birth. This de novo strategy is a direct recapitulation of naturally occurring human cancer-associated translocations. PMID- 12781365 TI - Altered receptor trafficking in Huntingtin Interacting Protein 1-transformed cells. AB - The clathrin-associated protein, Huntingtin Interacting Protein 1 (HIP1), is overexpressed in multiple human epithelial tumors. Here, we report that HIP1 is a novel oncoprotein that transforms cells. HIP1-transformed cells, in contrast to RasV12-transformed cells, have dysregulation of multiple receptors involved in clathrin trafficking. Examples include upregulation of the epidermal growth factor receptor (EGFR) and the transferrin receptor. Furthermore, accumulation of transferrin and EGF in the HIP1-transformed cells was increased, and breast tumors that had EGFR expressed also had HIP1 upregulated. Thus, HIP1 overexpression promotes tumor formation and is associated with a general alteration in receptor trafficking. HIP1 is the first endocytic protein to be directly implicated in tumor formation. PMID- 12781364 TI - PKC412 overcomes resistance to imatinib in a murine model of FIP1L1-PDGFRalpha induced myeloproliferative disease. AB - FIP1L1-PDGFRalpha causes hypereosinophilic syndrome (HES) and is inhibited by the tyrosine kinase inhibitor imatinib (Gleevec). Imatinib is a potent inhibitor of ABL, ARG, PDGFRalpha, PDGFRbeta, and KIT and induces durable hematologic responses in HES patients. However, we observed relapse with resistance to imatinib as consequence of a T674I mutation in FIP1L1-PDGFRalpha, analogous to the imatinib-resistant T315I mutation in BCR-ABL. We developed a murine bone marrow transplant model of FIP1L1-PDGFRalpha-induced myeloproliferative disease to evaluate the efficacy of PKC412, an alternative inhibitor of PDGFRalpha, for the treatment of HES. PKC412 is effective for treatment of FIP1L1-PDGFRalpha induced disease and of imatinib-induced resistance due to the T674I mutation. Our data establish PKC412 as molecularly targeted therapy for HES and other diseases expressing activated PDGFRalpha and demonstrate the potential of alternative kinase inhibitors to overcome resistance in target tyrosine kinases. PMID- 12781366 TI - Human mammary epithelial cell transformation through the activation of phosphatidylinositol 3-kinase. AB - Recent studies have demonstrated that introduction of hTERT in combination with SV40 large T antigen (LT), small t antigen (st), and H-rasV12 suffices to transform many primary human cells. In human mammary epithelial cells (HMECs) expressing elevated c-Myc, activated H-Ras is dispensable for anchorage independent growth. Using this system, we show that st activates the PI3K pathway and that constitutive PI3K signaling substitutes for st in transformation. Moreover, using constitutively active versions of Akt1 and Rac1, we show that these downstream pathways of PI3K synergize to achieve anchorage-independent growth. At lower levels of c-myc expression, activated PI3K also replaces st to complement H-rasV12 and LT and confers both soft agar growth and tumorigenicity. However, elevated c-myc expression cannot replace H-rasV12 for tumorigenesis. These observations begin to define the pathways perturbed during the transformation of HMECs. PMID- 12781368 TI - Caught up in a Wnt storm: Wnt signaling in cancer. AB - The Wnt signaling pathway, named for its most upstream ligands, the Wnts, is involved in various differentiation events during embryonic development and leads to tumor formation when aberrantly activated. Molecular studies have pinpointed activating mutations of the Wnt signaling pathway as the cause of approximately 90% of colorectal cancer (CRC), and somewhat less frequently in cancers at other sites, such as hepatocellular carcinoma (HCC). Ironically, Wnts themselves are only rarely involved in the activation of the pathway during carcinogenesis. Mutations mimicking Wnt stimulation-generally inactivating APC mutations or activating beta-catenin mutations-result in nuclear accumulation of beta-catenin which subsequently complexes with T-cell factor/lymphoid enhancing factor (TCF/LEF) transcription factors to activate gene transcription. Recent data identifying target genes has revealed a genetic program regulated by beta catenin/TCF controlling the transcription of a suite of genes promoting cellular proliferation and repressing differentiation during embryogenesis, carcinogenesis, and in the post-embryonic regulation of cell positioning in the intestinal crypts. This review considers the spectra of tumors arising from active Wnt signaling and attempts to place perspective on recent data that begin to elucidate the mechanisms prompting uncontrolled cell growth following induction of Wnt signaling. PMID- 12781367 TI - A peptide trivalent arsenical inhibits tumor angiogenesis by perturbing mitochondrial function in angiogenic endothelial cells. AB - Mitochondria are the powerhouse of the cell and their disruption leads to cell death. We have used a peptide trivalent arsenical, 4-(N-(S glutathionylacetyl)amino) phenylarsenoxide (GSAO), to inactivate the adenine nucleotide translocator (ANT) that exchanges matrix ATP for cytosolic ADP across the inner mitochondrial membrane and is the key component of the mitochondrial permeability transition pore (MPTP). GSAO triggered Ca(2+)-dependent MPTP opening by crosslinking Cys(160) and Cys(257) of ANT. GSAO treatment caused a concentration-dependent increase in superoxide levels, ATP depletion, mitochondrial depolarization, and apoptosis in proliferating, but not growth quiescent, endothelial cells. Endothelial cell proliferation drives new blood vessel formation, or angiogenesis. GSAO inhibited angiogenesis in the chick chorioallantoic membrane and in solid tumors in mice. Consequently, GSAO inhibited tumor growth in mice with no apparent toxicity at efficacious doses. PMID- 12781370 TI - 3rd Geneva aging workshop 2002: cancer, apoptosis and aging. AB - The topics of the 3rd Geneva Aging Workshop were cancer, apoptosis, and aging. As the risk for cancer is increasing with age, synergies that could emerge from this encounter could yield significant insights into the molecular mechanisms underlying both the etiology of cancer and the aging process. Several pathways that play a role in aging are thought to have an impact on the initiation and progression of cancer, with apoptosis being the key player. Which are the factors that are correlated with age-dependent cancer incidence? The regulators of apoptosis and genetic pathways that lead to apoptosis were discussed. This included genes that are clearly involved in aging, such as WS and BS, and genes that act in repair, such as BRCA1 and BARD1. The functions of telomere shortening and telomerase, extra cellular matrix, or epigenetic changes were presented in the light of their influence on aging and cancer risk. PMID- 12781369 TI - Raf proteins and cancer: B-Raf is identified as a mutational target. AB - A recent report has shown that activating mutations in the BRAF gene are present in a large percentage of human malignant melanomas and in a proportion of colon cancers. The vast majority of these mutations represent a single nucleotide change of T-A at nucleotide 1796 resulting in a valine to glutamic acid change at residue 599 within the activation segment of B-Raf. This exciting new discovery is the first time that a direct association between any RAF gene and human cancer has been reported. Raf proteins are also indirectly associated with cancer as effectors of activated Ras proteins, oncogenic forms of which are present in approximately one-third of all human cancers. BRAF and RAS mutations are rarely both present in the same cancers but the cancer types with BRAF mutations are similar to those with RAS mutations. This has been taken as evidence that the inappropriate regulation of the downstream ERKs (the p42/p44 MAP kinases) is a major contributing factor in the development of these cancers. Recent studies in mice with targeted mutations of the raf genes have confirmed that B-Raf is a far stronger activator of ERKs than its better studied Raf-1 homologue, even in cell types in which the protein is barely expressed. The explanation for this lies in a number of key differences in the regulation of B-Raf and Raf-1 activity. Constitutive phosphorylation of serine 445 of B-Raf leads to this protein having a higher basal kinase activity than Raf-1. Phosphorylation of threonine 598 and serine 601 within the activation loop of B-Raf at the plasma membrane also regulates its activity. The V599E mutation is thought to mimic these phosphorylations, resulting in a protein with high activity, leading to constitutive ERK activation. B-Raf now provides a critical new target to which drugs for treating malignant melanoma can be developed and, with this in mind, it is now important to gain clear insight into the biochemical properties of this relatively little characterised protein. PMID- 12781371 TI - Proceedings of the International Workshop on Taenia solium Cysticercosis/Taeniosis with Special Focus on Eastern and Southern Africa. Arusha, Tanzania, 19-22 August 2002. PMID- 12781372 TI - Background to the international action planning workshop on Taenia solium cysticercosis/taeniosis with special focus on Eastern and Southern Africa. PMID- 12781373 TI - Opening address by the Honorable Edward Lowassa (MP), Minister for Water and Livestock Development, Tanzania. PMID- 12781374 TI - The emergence of Taenia solium cysticercosis in Eastern and Southern Africa as a serious agricultural problem and public health risk. AB - Pig production has increased significantly in the Eastern and Southern Africa (ESA) region during the past decade, especially in rural, resource-poor, smallholder communities. Concurrent with the increase in smallholder pig keeping and pork consumption, there have been increasing reports of porcine cysticercosis in the ESA region. This article reviews the findings concerning the presence and impact of porcine cysticercosis in seven of the ESA countries. Most of the reported findings are based on surveys utilising lingual palpation and post mortem examination, however, some also used serological assays. In Tanzania, community-based studies on porcine cysticercosis indicate a prevalence of 17.4% in the northern highlands district of Mbulu and a prevalence range of 5.1-16.9% in the southern highlands. In Kenya recent surveys in the southwestern part of the country where smallholder pig keeping is popular indicate that 10-14% of pigs are positive for cysticercosis by lingual examination. Uganda has the most pigs in Eastern Africa, most of which are kept under smallholder conditions. Preliminary surveys in 1998 and 1999 at slaughterhouses in Kampala indicated a prevalence of porcine cysticercosis between 0.12 and 1.2%, however, a rural survey in northern Uganda in 1999 indicated 34-45% of pigs slaughtered in selected villages were infected. Additionally, a new survey of 297 pigs slaughtered in Kampala in 2002 indicated that pigs from the central region of the country were negative for cysticercosis while 33.7% of the pigs coming from the rural Lira district in the north were positive. Interestingly 8 piglet foetuses removed from an infected slaughtered sow coming from Lira district were all found to harbour cysts of T. solium providing evidence of congenital transmission of porcine cysticercosis. In Mozambique, abattoir records indicate that porcine cysticercosis is present in all provinces of the country. A serological survey on pigs in rural Tete Province found 15% of pigs positive. In Zimbabwe, a retrospective study in official abattoirs around the country from 1994 to 2001 reported a mean prevalence of 0.34% which is in contrast to a post-mortem survey in 1999, which showed that the prevalence of porcine cysticercosis in rural west Zimbabwe where smallholder pig keeping is popular was 28.6%. In Zambia, abattoir records reported porcine cysticercosis in six of the nine provinces. Routine meat inspection of 1316 pigs at a slaughter slab in Lusaka showed that 20.6% of the pigs had cysticercosis whereas serological testing of 874 pigs at the same abattoir indicated that 56.6% were found to have circulating antigens of Taenia solium. Field surveys based on lingual palpation in Southern and Eastern Provinces of Zambia revealed prevalences of 8.2-28.4 and 5.2%, respectively. South Africa has the largest number of pigs in Southern Africa and cysticercosis has been recognised as a problem in the country for many decades. There is strong evidence supporting the high prevalence of neurocysticercosis infecting humans from resource-poor areas of the country where pigs are being raised under smallholder conditions. In spite of this community-based surveys on porcine cysticercosis have never been conducted in South Africa and the last slaughterhouse survey was conducted nearly 40 years ago. The prevalences of porcine cysticercosis found in these ESA countries rank among the highest in the world and the disease is emerging as an important constraint for the nutritional and economic well being of resource-poor smallholder farming communities. The current findings suggest the widespread presence of human tapeworm carriers and thus a high risk of human cysticercosis in both rural areas and urban centres in the ESA region. More research is required in the region to assess the extent and public health and economic impact of T. solium infection in order to determine whether and what prevention and control efforts are needed. PMID- 12781375 TI - The current status of neurocysticercosis in Eastern and Southern Africa. AB - Some information has been documented on the epidemiology of neurocysticercosis in Eastern and Southern Africa through the monitoring of hospital-based patients with neurocysticercosis, community-based serological surveys of particular socio economic groups of people and surveys of porcine cysticercosis. Studies have revealed that non-pork eaters have as great a chance of infection as a pork eater, the Xhosa-speaking people of the Eastern Cape Province have the highest prevalence of cysticercosis/taeniosis in South Africa probably due to the common practice of free-range pig farming and the lack of sanitation in these areas. Several studies have revealed high prevalence rates in children and interestingly, patients with active cysts suffering from epilepsy. A startling mode of transmission is where self-trained healers use Taenia segments either for benevolent (e.g. in the treatment of severe intestinal tapeworm infections) or malevolent (evil) purposes (e.g. women "poisoning" an unfaithful husband or lover by adding the contents of Taenia solium segments to beer). PMID- 12781376 TI - Regional status, epidemiology and impact of Taenia solium cysticercosis in Western and Central Africa. AB - In West Africa, Taenia solium cysticercosis in both pigs and man has been reported in Benin, Burkina-Faso, Ghana, Ivory Coast, Senegal and Togo, and although official data are lacking, T. solium is anticipated to be present in most of the pig-raising regions of other West African countries as well. In some regions of Nigeria, the prevalence of porcine cysticercosis and human taeniosis is quite high (20.5 and 8.6%, respectively). Surprisingly, however, no cases of human cysticercosis have been reported, although epilepsy is very common. Large epidemiological surveys have only been carried out in Togo and Benin, where the prevalence of human cysticercosis was 2.4 and 1.3%, respectively. In Central Africa, porcine and human cysticercosis are (hyper)-endemic in Rwanda, Burundi, the Democratic Republic of Congo and Cameroon. The parasite also has been reported in pigs in Chad and Angola. Cysticercosis has been shown to be one of the major causes of epilepsy in Cameroon with figures as high as 44.6%. Cameroon is one of the few countries where the taeniosis-cysticercosis complex has been examined more in detail. In the Western province of Cameroon large scale surveys have shown that active cysticercosis is present in 0.4-3% of the local population and in 11% of the village pigs. However, the prevalence of adult T. solium was only 0.1%, which underscores the frequency of the T. solium paradox. Based on the available information, a very conservative economic estimate indicates that the annual losses due to porcine cysticercosis in 10 West and Central African countries amount to about 25 million Euro. The financial losses due to human cysticercosis are very difficult to estimate, but are certainly exceeded by the social impact of the disease, especially because of the particular perception of epilepsy in many African communities. It is concluded that the true prevalence of T. solium cysticercosis in pigs and humans in Central and West Africa remains underestimated because of unreliable slaughterhouse data and the lack of awareness and diagnostic facilities in the public health sector. PMID- 12781377 TI - Neurocysticercosis: regional status, epidemiology, impact and control measures in the Americas. AB - The analysis of epidemiological data concerning human cysticercosis point to important advances in understanding the magnitude and distribution of this parasitic disease in Latin America, as well as the relationship of the elements that conform the life cycle of Taenia solium. The data indicate that the main risk factor for acquiring human neurocysticercosis and swine cysticercosis is the presence of the tapeworm carrier in the household. Therefore, several intervention measures for the control of cysticercosis have been evaluated: mass treatment in order to cure tapeworm carriers, health education towards understanding the risk factors, pig control by restraining them, experimental vaccination of pigs and treatment of swine cysticercosis. In this paper, we review the information obtained in these areas. We hope it will be useful in other endemic countries that wish to elaborate an action plan for the control and ultimate eradication of T. solium. PMID- 12781378 TI - Taenia solium taeniosis/cysticercosis in Asia: epidemiology, impact and issues. AB - Several reports of patients with cysticercosis from many countries in Asia such as India, China, Indonesia, Thailand, Korea, Taiwan and Nepal are a clear indicator of the wide prevalence of Taenia solium cysticercosis and taeniosis in these and other Asian countries. However, epidemiological data from community based studies are sparse and available only for a few countries in Asia. Cysticercosis is the cause of epilepsy in up to 50% of Indian patients presenting with partial seizures. It is also a major cause of epilepsy in Bali (Indonesia), Vietnam and possibly China and Nepal. Seroprevalence studies indicate high rates of exposure to the parasite in several countries (Vietnam, China, Korea and Bali (Indonesia)) with rates ranging from 0.02 to 12.6%. Rates of taeniosis, as determined by stool examination for ova, have also been reported to range between 0.1 and 6% in the community in India, Vietnam, China, and Bali (Indonesia). An astonishingly high rate of taeniosis of 50% was reported from an area in Nepal populated by pig rearing farmers. In addition to poor sanitation, unhealthy pig rearing practices, low hygienic standards, unusual customs such as consumption of raw pork is an additional factor contributing to the spread of the disease in some communities of Asia. Undoubtedly, cysticercosis is a major public health problem in several Asian countries effecting several million people by not only causing neurological morbidity but also imposing economic hardship on impoverished populations. However, there are wide variations in the prevalence rates in different regions and different socio-economic groups in the same country. It is important to press for the recognition of cysticercosis as one of the major public health problems in Asia that needs to be tackled vigorously by the governments and public health authorities of the region. PMID- 12781379 TI - Human case management and treatment of cysticercosis. AB - Disease manifestations due to neurocysticercosis vary markedly and depend upon the location, size and number of cysts as well as the viability or degeneration of cysts and presence, type and degree of host response. Accordingly, the clinical management for each patient should be individualized. Treatment modalities include: (1) larvicidal drugs such as albendazole or praziquantel in patients with viable cysts; (2) corticosteroids or other agents to suppress or prevent the host's immune response; (3) anti-seizure medication(s) to treat or prevent recurrent seizures; (4) surgical interventions; and (5) the use of supportive family, social or health agencies in impaired individuals. Although it is known that larvicidal treatment kills viable cysts that commonly resolve or calcify, the clinical benefit of this treatment in the most common presentations is unproven. However, medical treatment of giant subarachnoid cysts, large parenchymal cysts or orbital cysts causing mass effect has led to definite clinical improvement in uncontrolled trials. Whether there is faster radiological improvement and/or clinical benefit in the treatment of cysts showing signs of inflammation by magnetic resonance imaging (enhancement and/or edema) is like wise controversial. There is no general understanding when or how to use corticosteroids to suppress natural or treatment-induced inflammation around cysts although their use when inflammation contributes or could be expected to cause undue morbidity or mortality is reasonable. Anti-seizure medication should usually be employed in patients with seizures or patients who may likely develop seizures. Surgical intervention is required to alleviate mass effect, remove some cysts causing obstruction of the ventricles, shunt placement for hydrocephalus, and sometimes for removal and/or decompression of large or critically located cysts before larvicidal treatment. PMID- 12781380 TI - Imaging findings in neurocysticercosis. AB - The diagnosis of neurocysticercosis was greatly improved by the introduction of computed tomography (CT) and magnetic resonance imaging (MRI). These techniques demonstrate the number and topography of lesions, their stage of involution, and the degree of inflammatory reaction of the host against the parasites and have largely replaced previous radiological procedures such as plain roentgenograms, pneumoencephalograms, cerebral angiography and myelography. In general, MRI provides better image detection and definition. The possibility of multiplanar reconstruction of images, its capability to visualize the posterior fossa without bone artifacts, and its high contrast resolution (far superior to that of CT) allow MRI to recognize many forms of cysticercosis not visualized on CT. However, the costs of MRI are high and the equipment is scarcely available in many endemic countries, and its sensitivity for the detection of calcified lesions is poor. CT remains the best screening neuroimaging procedure for patients with suspected neurocysticercosis, and MRI is the imaging modality of choice for the evaluation of patients with intraventricular cysticercosis, brainstem cysts and small cysts located over the convexity of cerebral hemispheres. Its better image definition also suggests that MRI is superior to CT in the follow-up of the patients after therapy. PMID- 12781381 TI - Immunodiagnostic tools for human and porcine cysticercosis. AB - The development of improved immunodiagnostic tools has contributed to our knowledge on the importance of taeniosis/cysticercosis by enabling sero epidemiological surveys and community-based studies to be carried out. Immunodiagnostic techniques include detection methods for specific antibodies and for circulating parasite antigen in serum or cerebrospinal fluid. The antigens used in immunoblot and enzyme-linked immunosorbent assay (ELISA) for antibody detection have evolved from crude extracts to highly purified specific fractions and recombinant antigens of the glycoprotein family, increasing both the sensitivity and the specificity of the tests. The application of ELISA for the detection of circulating parasite antigens may present some diagnostic advantages since it demonstrates not only exposure but also active infections. Until now only a few of the current techniques have been standardised and fully validated, making comparisons between studies difficult. The lack of a gold standard is a serious drawback. In surveys on cysticercosis, antibody detection systems have been useful in identifying the risk factors associated with transmission of Taenia solium; a high seroprevalence in a community indicates a "hot spot" where preventive and control measures should be applied. In contrast, the potential use of immunodiagnostic tools to identify cases of neurocysticercosis (NCC) in man is subject to debate. The correlation between a positive serology and neurological symptoms and/or lesions indicative for NCC on neuro-imaging techniques is poor to fair in most studies. This may be explained by the unpredictable clinical outcome of the infection and the variable immunological response of the human host to infection. A major problem is that in many developing countries, neuro-imaging methods are inaccessible and/or too expensive for the rural population at risk. Under these conditions, serology may provide the only tool for diagnosis of the infection. PMID- 12781382 TI - Immunodiagnostic tools for taeniasis. AB - Most diagnostic work conducted on the Taenia species zoonoses has been carried out on the larval stage of Taenia solium in man, reflecting the relative severity of the pathology caused by this stage of that organism. This review will, however, concentrate on the immunodiagnosis of the adult intestinal stages of these parasites in humans. Diagnosis of T. solium will be examined in most detail because of the relative importance of this parasite but relevant work from other cestodes of man and animals will also be discussed. In addition both classical and molecular approaches to diagnosis will be briefly covered. There have been a number of advances in immunodiagnosis of taeniasis over recent years that have improved both diagnostic sensitivity and specificity. Techniques for the detection of Taenia specific coproantigens in human taeniasis infections have been shown to more than double the numbers of T. solium cases accurately diagnosed in epidemiological studies. More recently, work on the serological diagnosis of T. solium have led to the development of a sensitive and specific enzyme linked immuno-transfer blot for the detection of species and stage specific circulating antibodies to adult worm excretory-secretory antigens. Work is ongoing to further improve these assays. PMID- 12781383 TI - Multiple genotypes of Taenia solium--ramifications for diagnosis, treatment and control. AB - Mitochondrial DNA sequences of Taenia solium have fully been analyzed. Analysis of the full length of cytochrome c oxidase subunit 1 (1620 bp) and cytochrome b (1068 bp) genes of T. solium, isolated from Asia (China, Thailand, Indonesia and India), from Latin America (Mexico, Ecuador, Bolivia, Peru and Brazil) and from Africa (Tanzania, Mozambique and Cameroon), has revealed that the two phylogenies obtained were similar to each other regardless of the genes examined. The isolates from Asia formed a single cluster, whereas those from Latin America combined with those from Africa to form an additional cluster. It was estimated that these two genotypes emerged approximately 4-8 x 10(5) years ago. These results together with recent study of the ancient of human taeniid cestodes emerged several MYA in Africa, historical data on swine domestication, distribution of pigs and colonization patterns suggest that T. solium was introduced recently into Latin America and Africa from different regions of Europe during the colonial age, which started 500 years ago, and that T. solium of another origin independently spread in Asian countries, perhaps from China. Why did not T. solium of European origin invade or spread into Asia during the colonial age? Analysis of T. solium distribution must include other Taenia species, especially T. saginata and T. asiatica, which can not be differentiated from each other morphologically. BESS T-base analysis for differentiation of all human Taenia species including the two genotypes of T. solium, and T. saginata and T. asiatica has also been characterized. BESS T-base analysis differentiates African isolates from Latin American isolates as well but more samples should be analyzed for obtaining conclusive evidence for the latter. Serological analysis of cyst fluid of T. solium cysticerci obtained in China and Indonesia and from Mozambique and Ecuador indicates geographical differences in their banding patterns. These differences are discussed in the light of possible differences in pathology of T. solium worldwide. As it has been speculated that the ancient T. solium emerged several million years ago in Africa, it is necessary to analyze more isolates from Africa. Such working hypothesis may be evaluated combined with symptomatology and serology when we get additional DNA data from such areas, since there are some varieties of manifestation of neurocysticercosis with or without subcutaneous cysticercosis and of antigens of cyst fluid of T. solium from Asia and from Africa and/or America. Transfer of techniques of molecular identification and sero- and immuno-diagnoses between researchers and technicians from endemic countries using their own materials should be promoted with the aim of better international cooperation for the control of cysticercosis. PMID- 12781384 TI - Control of Taenia solium. AB - Control or eradication of Taenia solium cysticercosis has been achieved to date only in Europe and North America. Significant improvements in sanitary conditions and developing functional slaughterhouse control systems were primarily responsible for control in these regions. Conversely, in endemic areas of developing countries control is limited by economic and sanitary conditions: the life cycle of T. solium is sustained because pigs have access to infected faeces, and cysticercosis-infested pork is available for consumption. Interventional trials with massive human cestocidal chemotherapy, treatment of both human and porcine populations with antihelminthic drugs and/or immunotherapy and health education have shown improvements in specific settings but not yet proven to be sustainable in the long-term. In order to ensure sustainability, any given control strategy towards elimination/eradication of porcine cysticercosis should incorporate economic incentives. PMID- 12781385 TI - Improving pig husbandry in tropical resource-poor communities and its potential to reduce risk of porcine cysticercosis. AB - To minimise the risk of cysticercosis in pigs it is necessary to raise pigs in confinement. The prevailing production system using free-range pigs is apparently very resilient although economic studies have shown that these traditional production systems are wasteful and unprofitable due to poor feed conversion, high mortality rates, low reproductive rates and poor final products. However, experiences from Africa show that intensive pig farming is stagnant and the sustainability of the traditional sector is better than that of the intensive sector. Examples from various African countries are given on the failure of intensive pig farming. This apparent paradox has various explanations. The lower fixed cost of traditional pig production compared with intensive production is one. Another explanation is that many households have some kind of kitchen waste, which can be exploited by a pig, but there is only enough for the partial feeding of a single pig. A third explanation of the paradox is that the pig has functions that are not reflected in a simple economic balance. The pig is a source of capital income, which can be realised at times of major expenses, and it can also be used as a way to put aside small amounts of money, which alternatively might evaporate. The pig's scavenging behaviour has clear nutritional benefits. Thus, a diet consisting of e.g. maize or sorghum, which are some of the feeds available on the small farms, will only provide approximately 30% of the pigs requirements of lysine and methionine, which are the most limiting amino-acids in pig feeds. Simulations with data available for green feeds and material of animal origin show that a 20% "supplement" from the fields may increase the amino-acid provision to about 80% of the optimum. If pigs are kept enclosed this supplement has to be fed to the pig which involves purchase and labour costs. The paper discusses the various options in terms of feeding, housing and use of genetic resources for the improvement of pig husbandry in tropical resource-poor countries. PMID- 12781386 TI - Improving meat inspection and control in resource-poor communities: the Nepal example. AB - Meat is an important source of protein and a valuable commodity in resource-poor communities. In many developing countries, lack of appropriate slaughtering facilities and unsatisfactory slaughtering techniques are causing unnecessary losses of meat as well as invaluable by-products from animal carcasses. Slaughtering places are frequently contaminated and may not be protected against dogs, rodents and insects. Meat products coming from such conditions are often deteriorated due to bacterial infection or contaminated, which may cause food poisoning or diseases in consumers. In many developing countries, regulations concerning meat inspection and/or control are inadequate or non-existent allowing consumers to be exposed to pathogens including zoonotic parasites. In Nepal, buffaloes contribute about 64% of the meat consumed, followed by goat meat (20%), pork (7%), poultry (6%) and mutton (2%). Goat and poultry meat is acceptable to all castes of people while buffalo meat is consumed mainly by the Newar ethnic group. Previously, pork was consumed only by people belonging to low castes, however, in recent years, the consumption of pork has increased in higher castes as the caste system has become more relaxed. Until recently, there were no official meat inspection regulations in the country, however, in 1999, the national government legislated an as-yet-to-be implemented Animal Slaughtering and Meat Inspection Act which mandates slaughterhouse construction and meat inspection and control. Due to the lack of implementation of the Meat Inspection Act and resultant absence of meat inspection, meat from sick or parasite-infected animals is serving as a source of infection to humans as well as other animals. In addition, meat quality is adversely affected by careless handling conditions in the slaughtering places as well as in the meat markets or shops. For improvement in animal slaughtering and meat inspection in both rural and urban areas of Nepal, several strategies are to be recommended. Sustainable capacity building should be introduced including training of veterinarians, meat inspectors and butchers as well as building of slaughter facilities. Government policies on slaughter procedures including ante-mortem examination, meat inspection and stamping of meat should be implemented. Programmes should be instituted with strong focus on prevention and control of meat-borne diseases to reduce infection risk of consumers and meat handlers and to avoid contamination of the environment. Lastly, emphasis should be put on improving the animal husbandry system in Nepal. These same actions can be undertaken in other developing countries to assist with improving meat inspection and control, thus helping with prevention and control of cysticercosis as well as other important meat-borne diseases. PMID- 12781387 TI - Vaccines for prevention of cysticercosis. AB - Neurocysticercosis due to Taenia solium infection is an important cause of human morbidity and mortality. Despite the availability of effective anthelmintics, the disease remains prevalent in many parts of the world and there is a need for new and improved measures for control of the infection. An effective vaccine to prevent infection in pigs, the parasite's natural intermediate host, would be a valuable new option to assist with T. solium control. Several approaches are being used currently towards the development of a T. solium vaccine and these approaches are reviewed briefly, with emphasis on the use of recombinant oncosphere antigens. Highly effective vaccines have been developed against cysticercosis in sheep and cattle caused by Taenia ovis and Taenia saginata, respectively. This success has encouraged the adoption of a similar strategy for T. solium. The recent finding that one oncosphere antigen, TSOL18, can induce complete protection against T. solium infection in pigs, highlights the potential for development of a practical vaccine. A vision is proposed for the development of a safe, effective, inexpensive vaccine for pigs, which can be administered in an edible form. Through an international collaborative effort, research is progressing towards the realisation of such a vaccine and its use to reduce the global burden of neurocysticercosis. PMID- 12781388 TI - Measures for the prevention and control of Taenia solium taeniosis and cysticercosis. AB - Taeniosis and cysticercosis due to Taenia solium are public health problems in many developing countries. Many studies of this parasitic zoonosis have focused on clinical features, diagnosis, treatment, surveillance, epidemiology and risk factors analysis. More recently projects on community and mass intervention strategies had been conducted in several rural areas worldwide focused on pig vaccination, pig cysticercosis treatment, human mass treatment, infrastructure development, as well as health education campaigns. Their advantages, disadvantages and public health impact have been published. This document discusses the feasibility and limitations of these interventions in order to assist countries in selection the best strategy for the prevention and control of this disease; we emphasized the specific strategies that might be recommended in different demographical situations. PMID- 12781389 TI - Taenia solium cysticercosis/taeniosis: potential linkage with FAO activities; FAO support possibilities. AB - Neurocysticercosis due to Taenia solium metacestodes is an important cause of human morbidity and mortality, particularly in parts of Latin America, Africa and Asia. The disease has been recognized as potentially eradicable. Emphasis has been placed on control through mass chemotherapy of human populations to remove tapeworm carriers, but this strategy does not control the source of infections, which is cysticercosis in pigs. Also, transmission may continue due to incomplete chemotherapy coverage of human carriers or because of immigration of tapeworm carriers into controlled areas. The FAO through the Veterinary Public Health (VPH) and Food Safety program has provided support for the write-up of guidelines for cysticercosis, diagnoses and control. This should be released in a joint effort with OIE and WHO and will provide regular support to seminars, workshops and congresses related to VPH. The FAO regular program has also established a global network of people directly involved in VPH, and is currently in the process of establishing four regional networks located in Asia, Africa, Eastern and Central Europe and Latin America. The networks should provide a basic framework to spread information related to diagnosis, prevention and control of major zoonotic diseases through electronic conferences, discussions, newsletters, and a Directory to establish contact with people involved in VPH and zoonotic diseases. Through the Technical Cooperation Program (TCP) the FAO has a tool to help Member Countries to create the basic environment to control emerging zoo sanitary problems, such as zoonotic and food borne diseases. PMID- 12781390 TI - The global campaign against epilepsy in Africa. AB - One of the leading brain disorders in developing countries is represented by epilepsy. It is estimated that 80% of people suffering from epilepsy around the world, reside in developing world such as Africa. Many perinatal and postnatal causes are brain-stressers in people suffering from malnutrition and low economical conditions. This context is characterised by long delay before modern treatment, reduced number and financial inaccessibility to anti-epileptic drugs (AEDs) and limited human and technical resources for epilepsy. Cultural interpretation also contributes to exclude epileptic patients from the educational and productive fields, aggravating the burden they face and favouring a treatment gap estimated to 80%. To fight against this dramatic reality, a partnership has been built between the International League against Epilepsy, the International Bureau for Epilepsy and the World Health Organisation, named the "Global Campaign Against Epilepsy" "Epilepsy Out of the Shadows" to reduce treatment gap and social and physical burden, educate health personnel, dispel stigma, support prevention. PMID- 12781391 TI - The US Centers for Disease Control and Prevention (CDC) and research and control of cysticercosis. AB - It is generally recognized that it is not possible to adequately protect the health of any nation without addressing infectious disease problems that occur elsewhere in the world. In 2002, the US Centers For Disease Control and Prevention (CDC) developed a revised strategy for consolidating, enhancing, and improving the effectiveness of CDC's efforts to prevent and control infectious diseases on a global scale. Taenia solium is one example of an imported infection disease, which impacts on the health of the US population but requires international coordinated efforts to prevent or limit transmission. This report outlines CDC's refocused global infectious disease strategy and how CDC collaborates in international efforts to eliminate taeniasis/cysticercosis. PMID- 12781392 TI - Using electronic technology for Taenia solium education: educating the educators. AB - In Taenia solium endemic areas, intervention programs include a variety of actions aimed to improve sanitary infrastructure and environment, pig husbandry conditions, detection and treatment of tapeworm carries and cysticercosis cases, among others. For a successful long-term effect, these actions must be associated with community participation and health education programs. Prior to designing/conducting health education programs tailored to specific populations, individuals involved in delivering education, as well as other professionals, students, and community leaders, must be well prepared, their knowledge increased, updated or strengthened. Educators must first be well educated to convey clear and accurate messages. Current electronic technology provides invaluable resources for teaching and learning. Learners using electronic materials benefit from a dynamic, non-linear and flexible interaction that can render long lasting effects compared to traditional passive methods. Electronic resources also provide the capability of producing one standardized template, versatile enough to be adapted, translated, modified, and updated. In this presentation, we introduce our preliminary work on the first interactive media presentation on T. solium which will hopefully, soon, become an interactive tutorial that could be utilized in health education in both developing and developed countries. PMID- 12781393 TI - Extension materials for meat-borne parasitic diseases in developing countries. AB - In support of a project on porcine cysticercosis in Tanzania, an educational video was prepared to inform the rural communities on the health risks and prevention of the parasitic disease. This paper describes the process involved in making the video, especially the importance of establishing a good understanding between veterinary public health officials and the video producer. Important steps in the process include determining the target audience, the film's core message, the construction of the "story", script development, the filming and editing activities, and, importantly, the development of strategies for production and use of the film as extension material. Suggestions on logistical and technical aspects of filming and viewing are also discussed. The experience gained in Tanzania will be of value to others planning similar projects elsewhere. PMID- 12781394 TI - The control of human (neuro)cysticercosis: which way forward? AB - Taenia solium cysticercosis, and its public health and economic consequences, appears to be a growing problem in poor areas of Africa, Asia and Latin America where people eat pork and traditional pig husbandry is practiced (and expanding). Its burden is counted in terms of human disease (mainly neurocysticercosis related epilepsy) and economic losses, in a context of both commercial and traditional subsistence pig farming. Although substantial fragmentary information seems to be available from local settings, national and global burdens due to T. solium cysticercosis are still to be comprehensively assessed. With regard to control, several strategies have been checked out at a small or medium scale and have proven to be successful. Yet, no intervention programmes have been implemented so far at the national level with proven success. Although T. solium cysticercosis is considered to be a potentially eradicable disease, there is no evidence yet that it is feasible and recommendable to envisage this within a reasonable time frame. However, it appears realistic to aim for the rapid definition of a simple package of interventions, which can routinely be carried out by existing services and structures, and will give an optimal, long-term return in terms of burden relief. Also, a number of international initiatives and opportunities currently exist in which a more pro-active attitude towards the control of T. solium cysticercosis can be integrated and promoted. Commitment of both national and local authorities to control the disease needs to be convincingly solicited and, as for most zoonotic diseases, an interdisciplinary approach is essential. PMID- 12781395 TI - Regional action plan for combating Taenia solium cysticercosis/taeniosis in Eastern and Southern Africa. PMID- 12781397 TI - Systematic reviews with individual patient data meta-analysis to evaluate diagnostic tests. AB - Systematic literature reviews with meta-analysis of reported data in diagnostic research studies about the accuracy of tests assist clinicians in decision making. However, there are limitations to this approach as the analysis of such data often does not allow reviewers to explore the diagnostic information gained from combinations of tests. In recent years meta-analysis of individual patient data (IPD) has been introduced as gold standard analytic approach in systematic reviews of randomised controlled trials. Application of IPD meta-analysis in systematic reviews of diagnostic tests can allow advanced analysis to decipher the real value of testing. In particular, the additional information provided by testing can be examined in light of the information already known from history and examination. To our knowledge there are no IPD meta-analyses in diagnostic research so far. This commentary aims to highlight the benefits of IPD meta analysis in the diagnostic domain and demonstrates how strategies for diagnostic work-up of women with postmenopausal bleeding could be improved using this approach in systematic reviews of diagnostic research on accuracy of ultrasound and hysteroscopy. PMID- 12781398 TI - Cervical cerclage in delayed interval delivery in a multifetal pregnancy: a review of seven case series. AB - OBJECTIVE: To examine whether cervical cerclage after the first delivery prolongs the inter-delivery interval in delayed interval deliveries. STUDY DESIGN: We identified 66 case reports and case series of delayed interval delivery published between 1880 and 2002. We selected seven case series that identified all cases of delayed interval delivery in their institutions during a specified period. RESULTS: Despite routine use of broad-spectrum prophylactic antibiotics, the average incidence of clinical intrauterine infection after the first delivery was 36% (95% confidence interval (CI): 26-46%). The incidence of maternal sepsis was 4.9% (95% CI: 0.2-9.6%). Studies in which cerclage was infrequently used reported a shorter inter-delivery interval compared to studies where cerclage was used in all cases (median is equal to 9 days versus 26 days, respectively, P<0.001) despite similar gestational ages at the first delivery, types of antibiotics, tocolytics, and incidence of infection. After controlling for other factors, the use of cerclage did not significantly increase the risk of intrauterine infection (adjusted relative risk=1.1, 95% CI: 0.4-3.5). CONCLUSION: Cervical cerclage after the first delivery is associated with a longer inter-delivery interval without increasing the risk of intrauterine infection. PMID- 12781399 TI - The number of abnormal plasma glucose values in the oral glucose tolerance test and the feto-maternal outcome of pregnancy. AB - OBJECTIVE: With respect to the feto-maternal outcome of pregnancy both appropriate screening and treatment of gestational diabetes mellitus (GDM) remain a matter of debate. Furthermore, the importance of only one abnormal glucose value at the glucose tolerance test (GTT) is still discussed, including the term "gestational impaired glucose tolerance" with normal fasting but an increased 2h postprandial glucose value. STUDY DESIGN: We have evaluated the feto-maternal outcome of pregnancy in 152 women with abnormal glucose values during the oral 100g GTT. The data were analysed with respect to the number of abnormal GTT values and compared to age- and body mass index (BMI)-matched control groups including 304 women with normal GTT values. RESULTS: A positive family history of diabetes was found in 24.4% of the GDM women and in 16.4% of the control group. In 45.9% of the women with abnormal GTT values glucose target levels could be obtained by dietary management, 54.1% required additional insulin therapy. Insulin therapy was administered in 32.8% of the women with one abnormal GTT value, in 65.0% of the women with two abnormal values and in 83.3% of the women with three abnormal values. Compared to the age- and BMI-matched control group the percentage of women with hypertension was increased in women with one, two and three abnormal GTT values. The percentage of LGA and macrosomic infants was significantly increased only in women with three abnormal GTT values. Women with one, two or three abnormal GTT values revealed an increased percentage of caesarean section compared to the control group. CONCLUSIONS: Our results show that women with three abnormal GTT values are at an overall increased risk with respect to the feto-maternal outcome of pregnancy. Compared to the control group also women with only one abnormal GTT value revealed an increased risk indicating a need for further control and therapy. PMID- 12781400 TI - HIV detection in amniotic fluid samples. Amniocentesis can be performed in HIV pregnant women? AB - OBJECTIVES: To assess amniotic fluid (AF) HIV contamination as a marker of intrauterine HIV infection and to evaluate amniocentesis as a risk factor for vertical HIV transmission when the mother was under antiretroviral treatment. STUDY DESIGN: Three hundred and sixty-six HIV pregnant women were included in the study. AF was obtained from three groups of patients: (a) genetic amniocentesis before 1997 (n=11); (b) amniocentesis a few days before the delivery day (n=18); and (c) AF collected on delivery (n=38). An univariate study was conducted to analyze amniocentesis as a risk factor of HIV transmission (groups a and b). Groups b and c were recruited after 1997; these patients were under combined antiretroviral treatment, they were studied to relate AF HIV contamination with fetal infection and maternal blood viral load at delivery (n=56). RESULTS: From 1984 to 1996, before antiretroviral therapy use in HIV pregnant women, transmission rate was 17%. In the group of patients who underwent genetic amniocentesis (group a) it was 30% (3/10) versus 16.2% (40/247) for patients without amniocentesis. Between 1997 and 2000 transmission rate was 3%. In group b it was 0% (0/18) when amniocentesis was done versus 3.7% (3/81) if no amniocentesis was done (no statistical differences). AF virus was undetectable in all samples (n=56) and no newborn infection was observed after the follow up. CONCLUSIONS: Amniotic fluid virus load was undetectable when maternal antiretroviral therapy was used, even if the virus was detectable in maternal blood samples. This finding raises the possibility to perform amniocentesis, when it is indicated, to provide the mother with an adequate antiretroviral treatment. PMID- 12781401 TI - Cord blood thyroid-stimulating hormone level in high-risk pregnancies. AB - OBJECTIVE: To evaluate the effect of various antepartum conditions on cord blood thyroid-stimulating hormone (TSH) level. STUDY DESIGN: The study group consisted of 24,892 consecutive singleton deliveries over a period of 4 years. The effect of preeclampsia, glucose intolerance, maternal medical diseases, and antepartum hemorrhage of unknown origin (APHUO) on cord blood TSH level were assessed by univariate analysis and linear regression. RESULTS: After controlling for potential confounders, there was a significant independent association between cord blood TSH level and preeclampsia (P=0.043), glucose intolerance (P=0.015), and maternal medical diseases (P=0.022). Antepartum hemorrhage of unknown origin was not associated with a higher cord blood TSH level. CONCLUSION: Cord blood TSH level was significantly elevated in various adverse antepartum conditions. This may be related to the placental insufficiency and fetal hypoxia commonly found in these high-risk pregnancies. PMID- 12781402 TI - Bacterial vaginosis: prevalence and predictive value for premature delivery and neonatal infection in women with preterm labour and intact membranes. AB - OBJECTIVES: Assess the predictive values of bacterial vaginosis (BV) for preterm delivery (PD) and neonatal infection and compare them with standard markers of infection among women with preterm labour (PL). STUDY DESIGN: Prospective blinded study in a tertiary referral centre in Paris. Women hospitalised for PL with intact membranes at a term between 24 and 34 weeks were included. Vaginal fluid, collected at inclusion was Gram-stained, scored, and interpreted according to Nugent's criteria. RESULTS: Out of 354 women tested, 254 had normal flora (72.3%), 76 intermediate (21.7%) and 24 BV (6.8%). A history of spontaneous miscarriage after 14 weeks was the only risk factor significantly associated with BV. BV was not significantly associated with PD<35 weeks or neonatal infection. Very preterm delivery (before 33 weeks) was significantly associated with the flora grade (P=0.02): women with normal, intermediate and abnormal flora, respectively had 27 (10.6%), 14 (18.4%) and 6 (25.0%) births before 33 weeks. Of the markers tested, the highest risk of very preterm delivery was associated with BV (odds ratio 2.95, 95% CI (1.1-0.8.1)) and CRP>20mg/dl (4.23 95% CI (1.8-9.7)). Predictive value of BV for preterm birth before 33 weeks were: sensitivity 12.8%, specificity 95.0%, positive predictive value 35.3%, and negative predictive value 84.3%. CONCLUSIONS: The frequency of BV and its association with PD are probably very variable and must be interpreted differently from one population to another. While we found an association between BV results and delivery before 33 weeks, the predictive value of BV was disappointing. Although these findings reinforce the importance of a useful marker of subclinical infection, the usefulness of testing for BV in women with PL has not been demonstrated. PMID- 12781403 TI - Effect of amniotic fluid upon prostaglandin E2 and I2 production by cultured human myometrial cells. AB - OBJECTIVE: Our goal was to study the effect of amniotic fluid obtained at 16 and 39 weeks of gestation in normal human pregnancies upon prostaglandin production by human myometrial cells in culture. STUDY DESIGN: Amniotic fluid, sampled either at 16 weeks, during amniocentesis, or at 39 weeks, during caesarean section before labor, was fractionated by molecular-weight and then incubated with human myometrial cells in culture. We then used radioimmunoassay to measure PGE(2) and PGI(2) production. RESULTS: The "3-30 kDa" fraction of amniotic fluid sampled at 16 weeks significantly inhibited PGE(2) and PGI(2) production by human myometrial cells. When amniotic fluid was sampled at 39 weeks, it stimulated both PGE(2) and PGI(2) production, and the ">30 kDa" fraction increased levels of PGE(2) considerably more than of PGI(2) (420.0+/-88.0 ng/10(6)cells versus 188.2+/-21.4 ng/10(6)cells, P<0.001). CONCLUSION: Amniotic fluid contains substances whose effects in cultured myometrial cells vary according to gestational age and type of prostaglandin. These data suggest that the fetus plays a role in the regulation of myometrial activity during pregnancy. PMID- 12781404 TI - A comparison of the neonatal morbidity of second twins to that of a low-risk population. AB - OBJECTIVE: To assess the neonatal morbidity of second twins. STUDY DESIGN: Cohort study in a department of perinatalogy. The neonatal morbidity of second twins was compared to that of a low-risk population: singletons in the cephalic presentation delivered vaginally. RESULTS: Five hundred fifty-nine second twins and 18,061 vaginally delivered singletons in the cephalic presentation were studied. Of 452 (81%) second twins delivered vaginally, 310 (69%) were extracted using obstetrical maneuvers: internal version and breech extraction, breech extraction alone, or assisted breech delivery if the breech was already engaged. Before 33 weeks of gestation, there was no significant difference between the neonatal morbidity of the vaginally delivered second twins and the vaginally delivered singletons in the cephalic presentation. After 33 weeks of gestation, only the 1-min Apgar score <7 and the rate of intubation at birth were significantly higher in the second twins. Whatever the gestational age, there was no significant difference between the neonatal morbidity of the vaginally delivered second twins and that of the second twins born by cesarean section before labor. At comparable gestational ages, there was no significant difference between the death rate of the vaginally delivered second twins and that in the reference population. CONCLUSION: The neonatal morbidity of second twins was comparable to that of a low-risk population. Immediate management of the vaginally delivered second twins was, however, more intensive than that of vaginally delivered singletons in the cephalic presentation. It, therefore, requires appropriate equipment in a suitable obstetric-pediatric setting. PMID- 12781405 TI - Placental inflammation and perinatal outcome. AB - OBJECTIVE: To examine the role of placental inflammation in adverse obstetrical outcome (AOO). METHODS: Analysis of perinatal data of 701 randomly selected mothers of singleton infants, Mombasa, Kenya. RESULTS: There were 661 (94.3%) live infants and 40 (5.7%) stillbirths. Out of the live born infants, 78 (12.4%) had a low birth weight (LBW < 2500g); 33 of them were preterm and 41 small for gestational age (SGA). The incidence of neonatal sepsis and post partum endometritis was 3.6 and 19.8%, respectively. The perinatal death rate was estimated to be 7.3% (51/701). The prevalence of acute placental inflammation was 19.6%. Acute placental inflammation was independently associated with preterm low birth weight (ARR=3.8, 95% CI=1.7-8.9, P<0.01), stillbirth (ARR=2.3, 95% CI=1.1 5.0, P=0.03) and perinatal death (ARR=2.8, 95% CI=1.4-5.4, P<0.01). Women with acute placental inflammation had a two-fold higher risk for AOO (32.6 versus 15.2%, respectively, ARR=2.5, 95% CI=1.3-4.8, P<0.01). Other risk factors for AOO were bad obstetrical history, low haemoglobin level and leucocytosis. CONCLUSIONS: The incidence of adverse obstetrical outcome defined as low birth weight, low Apgar score, perinatal mortality and post partum endometritis, was high in this population. Acute placental inflammation was associated with preterm birth, stillbirth and perinatal death. More research is needed to study the role of infection in adverse obstetrical outcome, and to design interventions to decrease infectious morbidity and mortality in pregnancy. PMID- 12781406 TI - Familial Mediterranean fever and its implications for fertility and pregnancy. AB - Familial Mediterranean fever (FMF) is a recessively inherited disease of episodic fever in combination with severe abdominal pain, pleurisy, arthritis or erysipelas-like skin rashes. The disease is mainly prevalent in Sephardic Jews, Armenians, Turks and Arabs. The gene responsible for FMF was cloned in 1997. The gene expresses a protein called pyrin which is believed to play a role in the downregulation of mediators of inflammation. Several mutations have been identified of which the homozygous form of the M694V mutation is associated with a more severe expression of FMF. Prophylactic administration of colchicine is effective in relieving most patients from their attacks and preventing the development of amyloidosis, which usually leads to end-stage renal disease. Unfortunately, there is little awareness of the disease in gynaecological practice although a FMF full blown episode may mimic an acute abdominal calamity suggesting several possible gynaecological diagnoses. FMF is also associated with subfertility. In females, infertility was mainly related to oligomenorrhea although the causes remain unclear. In male FMF patients, progression of the disease may induce testicular impairment, consequently affecting spermatogenesis. Some controversy exists as to the adverse effects of colchicine on sperm production and function although the impression is that the occurrence of sperm pathology in FMF patients, using the recommended dosage of colchicine, is very low. In pregnant FMF patients, an increased occurrence of miscarriage has been found. However, the mechanisms involved remain unclear. Although colchicine is a mitotic inhibitor and transplacental crossing of colchicine has been demonstrated, no increased risk of foetal abnormalities in colchicine-treated pregnant FMF patients has been found. Therefore, amniocentesis should not be done for reassurance alone. PMID- 12781407 TI - The involvement of progesterone in the onset of human labour. AB - Progesterone is essential for the maintenance of pregnancy, and acts to promote uterine quiescence. In many species progesterone levels fall prior to the onset of labour but no fall has been observed in humans. The concept of 'functional' progesterone withdrawal in association with labour has been proposed and a number of mechanisms have been suggested. Recent potential mechanisms of functional withdrawal include changes in progesterone receptor (PR) isoforms, regulation of PR responsive genes through nuclear factor kappaB (NF-kappaB) promoter sites and the non-genomic effects of progesterone or its metabolites. PMID- 12781408 TI - Successful pregnancies after using immotile spermatozoa from ejaculate, epididymis and testis. AB - OBJECTIVE: To assess the efficacy of intracytoplasmic sperm injection (ICSI) in term of pregnancy rate with immotile spermatozoa from ejaculate, epididymis and testis. STUDY DESIGN: A retrospective study was conducted between January 1998 and March 2001. We performed intracytoplasmic sperm injection with immotile spermatozoa, in 160 couples during 172 cycles. RESULTS: The birth rate per cycle was 38.4% in immotile spermatozoa from ejaculate, 35.4% from testis and 38.7% from epididymis. CONCLUSION: This retrospective analysis shows that immotile spermatozoa retrieved from epididymis or testicle gives similar fertilization and pregnancies rates as immotile spermatozoa from ejaculate. PMID- 12781409 TI - Cryopreservation of human ovarian tissue by direct plunging into liquid nitrogen. AB - AIM: To establish a prospective direction for further development of the protocol for cryopreservation of ovarian tissue by direct plunging into liquid nitrogen. MATERIALS AND METHODS: Human ovarian biopsies from 20 patients (cut in approximately 0.5mm(3) pieces) were exposed to: 40% ethylene glycol+0.35 M sucrose+5% egg yolk; 40% ethylene glycol+18% Ficoll-70+0.35 M sucrose; 20% ethylene glycol+20% dimethyl sulphoxide. Cryopreservation of pieces was accomplished by plunging 0.25 ml straws or copper grids into liquid nitrogen or 0.25 ml straws into precooled (-196 degrees C) metallic powder. Thawed pieces were transferred to sucrose solution for incremental dilution of cryoprotectants. Histological observation of the tissue was performed after cryopreservation and in vitro culture was done to study hormone production ability after cryopreservation. RESULTS: Only ultrarapid cooling in ethylene glycol-sucrose-egg yolk solution protected both follicles and stroma from damage. CONCLUSION: The following parameters were established as required for a protocol of human ovarian tissue cryopreservation by direct plunging into liquid nitrogen: the vitrification medium should include ethylene glycol, disaccharide and egg yolk; ultrarapid cooling/thawing should take place using standard 0.25 straws or copper grids. PMID- 12781410 TI - Pregnancy outcome after laparoscopic and laparoconverted myomectomy. AB - OBJECTIVE: To compare fertility and pregnancy-related complications after laparoscopic and laparoconverted myomectomy. METHODS: Retrospective analysis of 106 infertile women with uterine leiomyomas, of whom 88 women underwent laparoscopic myomectomy and 18 laparoconversion. RESULTS: There was no difference in the patients' baseline characteristics or the mean number of fibroids between the laparoscopic and laparoconversion groups. The mean (+/-S.D.) diameter of the largest fibroid in the laparoscopic and laparoconversion groups was 6.2+/-1.8 and 8.1+/-1.4 cm, respectively (P<0.001). There was no difference in operating time between the two groups. The hospital stay was shorter in the laparoscopic group: 3.0+/-1 versus 5.5+/-1 days (P<0.001). The mean follow-up in the laparoscopic and laparoconversion groups was 27.3+/-7.0 and 32.0+/-3.1 months, respectively (NS). No difference in the pregnancy rate was noted between the laparoscopic and laparoconversion groups (48 and 56%, respectively). The mean time before conception in the laparoscopic and laparoconversion groups was 7.5+/-2.6 and 15.1+/-2.4 months, respectively (P<0.001). There was no difference between the two groups as regards the rates of pregnancy-related complications and vaginal delivery. No uterine rupture occurred. CONCLUSION: Laparoscopic myomectomy is feasible and safe, and should be considered for infertile women with uterine fibroids. Fertility and pregnancy outcomes following laparoscopic myomectomy are comparable with those following myomectomy after laparoconversion. PMID- 12781411 TI - Glycodelin serum levels in women with ectopic pregnancy. AB - OBJECTIVE: Aim of our study was to determine circulating levels of glycodelin for biochemical monitoring of women with vaginal bleeding and/or abdominal pain in early pregnancy. The objective was, using glycodelin as a biochemical parameter, to distinguish between incomplete abortion and ectopic pregnancy in early weeks of gestation. STUDY DESIGN: In 169 women with a first trimester pregnancy, a single serum measurement of maternal glycodelin was taken. Patients were divided into groups according to the clinical and/or ultrasonografic findings at the time of hospitalisation:ectopic pregnancy, incomplete abortion and control. RESULTS: Glycodelin serum levels were significantly lower in patients with ectopic pregnancy comparable with intact pregnancy and incomplete abortion. There was no difference in serum levels between intact pregnancy and incomplete abortion. CONCLUSION: Glycodelin might represent a biochemical parameter in the differential diagnosis between ectopic pregnancies and incomplete abortion. The number of patients was too small to give reference ranges for pregnancy weeks. PMID- 12781412 TI - Radioactivity in cervical mucus after intraperitoneal deposition of Tc-99m labelled albumin particles before and after sterilisation. AB - OBJECTIVE: To study the transport of radiolabelled albumin particles from the pouch of Douglas to cervical mucus. STUDY DESIGN: A prospective randomised study, including 10 healthy women, undergoing laparoscopic sterilization. A solution of human serum albumin particles, labelled with Technetium 99(m) (Tc-99(m)) was injected into the pouch of Douglas, either before or after sterilisation. RESULTS: Gamma camera images showed gradual spread to the entire peritoneal cavity. Blood samples, showed increasing levels of radioactivity. Measurement of radioactivity in cervical mucus showed significant activity both in women sterilized before and after the deposition. CONCLUSION: The study showed evidence of transport of radioactivity from the pouch of Douglas to cervical mucus. The transport seemed to consist of free activity since no radionuclide labelled particles could be detected in cervical mucus. The transport was probably haematogenous or lymphatic and not intraluminal through the fallopian tube. Further studies utilizing particles comparable in size to fertilized ova are required to design methods for evaluating the transport from the fallopian tube to the uterine cavity. PMID- 12781413 TI - How significant is a cervical smear showing glandular dyskaryosis? AB - OBJECTIVE: To evaluate the incidence, outcome and predictive value of cytology showing glandular dyskaryosis. PARTICIPANTS: Fifty-seven women with a smear diagnosis of glandular dyskaryosis registered between January 1997 and December 2001. SETTING: Colposcopy and cytopathology units in a large district general hospital. RESULTS: Sixty smears in 57 women showing glandular dyskaryosis were identified from a cohort of 135,120 smears, giving an incidence of 0.05%. Hospital records were available for 50 women. Final diagnosis included 13 cases of cervical glandular intraepithelial neoplasia (CGIN), 4 microinvasive cervical adenocarcinomas, 2 undifferentiated tumours, 1 microinvasive squamous carcinoma, 21 cases of CIN and 13 cases of endometrial pathology (8 endometrial cancers). Twelve women had coexistent squamous and glandular disease. Forty-five out of 50 women had significant pathology (positive predictive value 90%). Colposcopy was seen to be of limited value in assessment of smears showing glandular dyskaryosis. Only 1 out of 13 glandular lesions was diagnosed by colposcopy. CONCLUSION: Smears showing glandular dyskaryosis are associated with significant pathology in 90% of cases and malignancy in 32% of cases. Hence, women with a smear showing glandular dyskaryosis should be referred urgently to a colposcopy clinic and flagged up as suspected cancer. Glandular dyskaryosis should be included in the national referral criteria for suspected gynaecological cancer. PMID- 12781414 TI - Clinical over- and under-estimation in patients who underwent hysterectomy for atypical endometrial hyperplasia diagnosed by endometrial biopsy: the predictive value of clinical parameters and diagnostic imaging. AB - OBJECTIVE: The aim of this study was to analyze the clinical and imaging characteristics in patients diagnosed with atypical endometrial hyperplasia based on endometrial biopsy in comparison with the final diagnosis from resected uteri; i.e. to determine the rates of underestimation (endometrial cancer), equivalent diagnosis (atypical hyperplasia), and overestimation (hyperplasia without atypia or non-hyperplastic lesion). MATERIALS AND METHODS: We reviewed 33 patients who were diagnosed with atypical endometrial hyperplasia by endometrial biopsy using a small curette and then underwent total abdominal hysterectomy between September 1992 and May 2002. Clinical parameters obtained from patients' charts, and imaging analyses using transvaginal ultrasonography (TUS) and magnetic resonance (MR) imaging were retrospectively re-examined. RESULTS: Among 33 patients who underwent hysterectomy due to a diagnosis of atypical hyperplasia, nine cases (27.2%) were underestimated (cancer), nine cases (27.2%) were equivalent and 15 cases (45.6%) were overestimated as indicated by examination of the endometrium of the resected uterus. There was no difference among these groups in either clinical parameters or diagnostic images obtained by TUS or MR. CONCLUSION: Diagnosis of atypical endometrial hyperplasia by endometrial biopsy often resulted in under- or over-estimation, as shown by examination after hysterectomy. As there is neither a reliable clinical parameter nor imaging feature to distinguish between these groups, hysterectomy is still the best treatment for these patients if they are willing to give up their fertility. PMID- 12781415 TI - Laparoscopic management of adnexal masses in pregnancy: a case series. AB - OBJECTIVE: To determine the feasibility, safety, limiting factors, and advantages of laparoscopic management of adnexal masses in pregnancy. STUDY DESIGN: During a 12-year period, 48 laparoscopic procedures were performed in 47 patients with adnexal masses in pregnancy. Laparoscopic surgery was done during the first trimester of pregnancy in 17 cases, the second trimester in 27 cases and the third trimester in four cases. All the procedures were performed with general anesthesia and curarization. The laparoscopic cystectomies were performed either with the intra-peritoneal or the trans-peritoneal technique. RESULTS: The indications were: persistant or sonographically abnormal ovarian cyst (36 cases), torsion or rupture of ovarian cyst (8 cases), and symptomatic pelvic mass (3 cases). Two borderline tumors were discovered. The laproscopic procedure could not be performed in two cases due to dense adhesions and difficulty of hemostasis. No patient encountered complications during the intra- and post operative periods. The mean hospital stay was 3.8 days. The outcome of the pregnancy was normal in all cases except one fetal loss 4 days after the laparoscopy. CONCLUSION: Laparoscopic management of adnexal masses in pregnancy by an experienced team, is a safe and effective procedure that allows, compared to the traditional surgery, a shorter hospital stay, a reduced rate of post operative complications and a decreased maternal and fetal morbidity. PMID- 12781417 TI - Congenital pelvic arteriovenous malformation: two cases and MR findings. AB - Congenital pelvic arteriovenous malformation (PAVM) is rare. We present two cases with emphasis on the imaging features. Magnetic resonance angiography (MRA) demonstrates comparable details of the vascular abnormalities as conventional angiography, which should be reserved for pre-embolization evaluation. PMID- 12781416 TI - Huge maternal hydronephrosis: a rare complication in pregnancy. AB - A huge maternal hydronephrosis is uncommon in pregnancy and might be mistaken as a pelvic mass. A 21-year-old primigravida was noted at 25th week of gestation to have a visible bulging mass on her left flank. The mass was originally mistaken as a large ovarian cyst but later proved to be a huge hydronephrosis. Retrograde insertion of ureteroscope and a ureteric stent failed, so we performed repeated ultrasound-guided needle aspiration to decompress the huge hydronephrosis, which enabled the patient to proceed to a successful term vaginal delivery. Nephrectomy was performed after delivery and proved the diagnosis of congenital ureteropelvic junction obstruction. PMID- 12781418 TI - Garnerella vaginalis bacteremia after vaginal myomectomy. PMID- 12781419 TI - Partial necrosis of cervix uteri as a complication to HELLP-syndrome. AB - BACKGROUND: The HELLP-syndrome is a rare complication to pregnancy and a potential fatal condition. CASE: We report such a case in a 32-year-old para 2, gravida 1 woman with no known risk factors. A lower transverse uterotomy was performed at 29 weeks and 6 days' gestation based on an indication of HELLP syndrome. Uncharacteristically lower back pain developed on the second day postoperatively and a gynaecological examination was performed. A partial necrotic cervix uteri was found with many trombosed vessels and necrotic tissue. CONCLUSION: Partial necrosis of cervix uteri can be seen as a complication to the HELLP-syndrome. PMID- 12781420 TI - Specialist life--Jim Thornton. PMID- 12781421 TI - Treatment of relapsed small-cell lung cancer--a focus on the evolving role of topotecan. AB - Despite the high response rates to chemotherapy, small-cell lung cancer (SCLC) is among the most lethal malignancies. Long-term survival is anecdotal for patients with extensive disease; 5-years survival is < or =5% for those with limited disease. All patients with extensive disease and most patients with limited disease will experience disease progression and become candidates for second-line therapy. Although a number of agents have demonstrated antitumor activity in relapsed SCLC, including paclitaxel, docetaxel, etoposide, cisplatin, and carboplatin, topotecan is the only single agent currently approved in the United States for the treatment of recurrent disease. Topotecan is a novel topoisomerase I inhibitor with established antitumor activity in recurrent SCLC and has a predictable, noncumulative toxicity profile. Furthermore, topotecan has been shown to provide symptom improvement in this predominantly palliative setting. Evidence also suggests that topotecan readily penetrates the blood-brain barrier and might be active in the relatively large subset of SCLC patients who experience brain metastases. This article reviews the clinical utility of topotecan in recurrent SCLC, including its efficacy, tolerability, and quality-of life effect, when used as monotherapy and in novel combination regimens. PMID- 12781422 TI - Survival among Chinese women with lung cancer in Singapore: a comparison by stage, histology and smoking status. AB - Lung cancer generally carries a poor prognosis and the determinants of survival have been of interest. However, survival estimates in Asian populations are scarce. This study describes survival rates and their determinants in Singapore Chinese women, a primarily non-smoking population. Three hundred and twenty-six Chinese women, diagnosed with primary lung carcinoma in three major hospitals in Singapore between April 1996 and December 1998, were followed up till 31 December 2000. The Kaplan-Meier method was used for survival analysis. Two hundred and eighty (85.7%) died from the disease during follow-up. The median survival time was 0.7 years and the three-year survival was 15.8%. These survival rates are similar to those of Western populations, and they provide a basis for examining trends over time. Age at diagnosis was an independent prognostic factor [adjusted hazard ratio (relative risk) 1.4, 95% confidence intervals (CI) 1.1-1.9 for women above 65 years relative to younger women]. Most (70.5%) tumours were stage III/IV at diagnosis. Three-year survival ranged from 72% among patients with stage I tumours to 7% for stage IV tumours. Overall, there was no survival difference among different histological types in all stages combined. When limited to stages I and II cancers, adenocarcinomas were associated with a better outcome relative to other histological subtypes combined (adjusted relative risk 0.4, 95% CI 0.1 1.0). Smoking was an independent risk factor (adjusted relative risk 1.3, 95% CI 1.0-1.8). Nevertheless, non-smokers comprised 57.4% of this series, highlighting the importance of increased awareness among health professionals and the public that lung cancer is not only a disease of smokers. The high proportion of late stage tumours in this study and the impact of disease stage on outcome underline the importance of early detection in improving survival of lung cancer. PMID- 12781423 TI - A population-based study of glutathione S-transferase M1, T1 and P1 genotypes and risk for lung cancer. AB - A deletion polymorphism for glutathione S-transferase M1 (GSTM1) has been related to risk for lung cancer among smokers in some studies but not in others. We examined GSTM1, a GSTT1 deletion polymorphism and a common GSTP1 gene variant (iso-->val), as risk factors for lung cancer in a population-based case-control study of men. Cases (N=274) were males identified from 1993 to 1996 through the Fred Hutchinson Cancer Research Center Cancer Surveillance System registry for western Washington State. Male age-matched controls (N=501) were selected by random-digit dialing. Subjects participated in a telephone interview and blood draw. GSTM1 and GSTT1 were genotyped with a multiplex PCR assay using beta-globin as a positive control, and GSTP1 single nucleotide variant determined with PCR based oligonucleotide ligation assays. GSTM1 absence was associated with a modest elevation in risk among all cases (odds ratio=1.27, 95% CI 0.91-1.77) and among non-small cell cancers (adenocarcinoma OR=1.58, 95% CI 0.99-2.52; squamous cell OR=1.40, 95% CI 0.83-2.34). Risk associated with GSTM1 null was increased two to sixfold among heavy smokers. GSTT1 was not associated with lung cancer risk and GSTP1 val was non-significantly associated with a modest reduction in risk, particularly among heavy smokers. No specific combination of GST genotypes was particularly associated with risk. These results support previous reports that the GSTM1 null genotype is associated with a modest increase in risk for lung cancer, particularly among heavy smokers, suggest no role for GSTT1 and the need for further study of GSTP1. PMID- 12781424 TI - P53 (codon 72) and P21 (codon 31) polymorphisms alter in vivo mRNA expression of p21. AB - p21 (Waf1/Cip1) is a downstream target of p53. We evaluated the association between p21 polymorphism (codon 31), p53 polymorphism (codon 72) and their corresponding in vivo mRNA expression. In this study, p21 and p53 genetic polymorphisms (using standard PCR-RFLP techniques) and p21 and p53 gene expressions (using a radiolabelled ribonuclease protection assay (RPA) technique) were evaluated in the peripheral leukocytes of 84 individuals (63 with lung cancer). Log-transformed values of mRNA expression by RPA, which approximated a normal distribution, were analyzed. p53 genotypes did not correlate with p53 mRNA log-expression (P>0.05 for all comparisons), but the Pro allele variants of p53 were associated with a significant decrease in mRNA log-expression of its downstream target, p21. The variant Arg allele of p21 was also associated with a significant decrease in p21 mRNA log-expression. When individuals with at least one variant allele of both p53 and p21 (double-variants) were compared with all other genotype groups, these double-variants had significantly lower log expression of p21 (P<0.005 by both t-tests (crude) and linear regression analyses (adjusted)). This is translated into an approximate 48% reduction in the geometric mean of the mRNA expression of the double-variants, when compared with all other groups. Results were consistent in both patients with lung cancer (n=63) and in normal controls (n=21). In conclusion, the presence of a p53 Pro allele and/or p21 Arg allele is associated with lower downstream target gene expression of p21. PMID- 12781425 TI - Discovery of distinct protein profiles specific for lung tumors and pre-malignant lung lesions by SELDI mass spectrometry. AB - OBJECTIVES: Early lung cancer detection and treatment remain a challenge. The efficacy of surface-enhanced laser desorption/ionization (SELDI) technology in lung cancer detection, has not been defined. This study identifies specific protein peak patterns in malignant lung tumors, and in pre-malignant airways epithelium showing neoplastic transformation. METHODS: Lung tumor specimens taken from patients participating in a lung cancer screening study (H. Lee Moffitt Cancer Center, Tampa, FL) were laser capture microdissected to obtain pure cell populations from frozen sections of normal lung, atypical adenomatous hyperplasia (AAH) and malignant tumors. SELDI mass spectrometry was used to generate protein profiles in each epithelial cell type. RESULTS: SELDI mass spectroscopy is highly reproducible in detecting lung tumor-specific protein profiles. Three peaks at 17 23 kDa mass range from tumor cells showed markedly increased compared with normal cells. The peak at 17250 Da was not detected in any of the normal cells. This peak appeared to be present at low levels in the atypical cell samples. CONCLUSIONS: This study demonstrates the feasibility of detecting "malignant" protein signatures from lung tumor and pre-malignant pulmonary epithelium using SELDI mass spectrometry. Although additional study is necessary to validate these patterns as unique diagnostic tools, these "malignant" protein signatures lend themselves to identification of populations at high-risk for lung cancer and for monitoring response to lung cancer chemopreventive agents. PMID- 12781426 TI - Mutational analysis of the transforming growth factor beta receptor type I gene in primary non-small cell lung cancer. AB - Transforming growth factor-beta receptor-dependent signals are critical for cell growth and differentiation and are often disrupted during tumorigenesis. The entire coding region of TGFbetaRI and flanking intron sequences from 53 primary non-small cell lung cancer (NSCLC) tissues were examined for alterations using SSCP and direct sequencing. No somatic point mutations other than two silent mutations and a polymorphism were found in the TGFbetaRI gene. The two silent mutations located at codon 344 (AAT to AAC) and codon 406 (TTA to CTA), respectively, and the polymorphism was at the 24th base of intron 7 (G to A). To investigate whether the presence of this polymorphism is associated with NSCLC, we determined its allele distribution in all the 53 carcinomas and 89 normal controls. Interestingly, we found that the subjects with homozygous genotype A/A displayed more than 3-fold increased risk of developing NSCLC than the common wild genotype G/G. As the first report, the present study showed that TGFbetaRI gene is not a frequent site of spontaneous mutational inactivation while the detected polymorphism is frequent in the pathogenesis of NSCLC. PMID- 12781427 TI - Diagnostic value of CA 549 in pleural fluid. Comparison with CEA, CA 15.3 and CA 72.4. AB - Several tumor markers have been evaluated in pleural fluid, but their clinical role has not been firmly established. The aim of this study is to determine the diagnostic value of carbohydrate antigen 549 (CA 549) levels in pleural fluid, and to compare it with another previously studied tumor markers: carcinoembryonic antigen (CEA), CA 15.3 and CA 72.4. We prospectively studied 252 patients with pleural effusion: 101 malignant (20 mesothelioma) and 151 of several benign diseases. The levels of the tumor markers were measured by immunoradiometric assays (RIA). CA 549 in pleural fluid has an acceptable sensitivity (0.49), with high specificity (0.99). The best combination of tumor markers for differentiating malignant from benign effusions was CA 549+CEA+CA 15.3, with a sensitivity of 0.65, specificity of 0.99 and accuracy of 0.85. The addition of any one tumor marker assay consistently improved the diagnostic value of cytology. In our study, none of the tumor markers was organ-specific. When mesothelioma and hematological malignancy were ruled-out, the combination of CA 549+CEA+CA 15.3, improved the results up to a sensitivity of 0.77, specificity of 1 and accuracy of 0.92. In conclusion, CA 549 assay has an acceptable sensitivity with high specificity. The best combination of tumor markers in this series with a high relative frequency of mesothelioma and low frequency of breast carcinoma was CA 549+CEA+CA 15.3. Individual tumor markers or their combination increased the sensitivity of pleural cytology. PMID- 12781428 TI - A prospective study of the impact of weight loss and the systemic inflammatory response on quality of life in patients with inoperable non-small cell lung cancer. AB - The relationship between weight loss, the systemic inflammatory response and quality of life in patients with inoperable non-small cell lung cancer (NSCLC) was studied. The extent of weight loss, the systemic inflammatory response (C reactive protein) and quality of life (EORTC-QLQ-C30) was measured in 106 patients with inoperable NSCLC (stage III and IV). Approximately 40% had more than 5% weight loss and almost 80% had elevated circulating C-reactive protein concentrations (>10 mg/l). The functional scale scores of the EORTC-QLQ-C30 were poor (50 or less) and the fatigue symptom score was also poor (50 or more). When patients were grouped according to whether or not they had experienced more than 5% weight loss, Karnofsky performance status and global quality of life were lower (P<0.05) and symptom scores fatigue (P<0.05) and pain (P<0.01) were greater in the weight-losing group. When the weight-stable cancer patients were grouped according to whether or not they had evidence of a systemic inflammatory response, the symptom fatigue was higher in the inflammatory group (P<0.05). In the weight-stable cancer patients C-reactive protein concentration was correlated with fatigue r=0.31 (P<0.05). The results of the present study indicate that both weight loss and the systemic inflammatory response impact on different aspects of quality of life. In particular, fatigue is associated with the presence of a systemic inflammatory response independent of weight loss. PMID- 12781429 TI - ZD1839, a novel, oral epidermal growth factor receptor-tyrosine kinase inhibitor, as salvage treatment in patients with advanced non-small cell lung cancer. Experience from a single center participating in a compassionate use program. AB - OBJECTIVE: We evaluated the efficacy and tolerability of the orally active, selective epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) ZD1839 in patients with pretreated advanced non-small cell lung cancer (NSCLC) participating in a compassionate use program. PATIENTS AND METHODS: Thirty-one patients with advanced, unresectable and progressive NSCLC, previously treated with one or two chemotherapy regimens, received ZD1839 250 mg orally once daily. Patients who had received only one prior chemotherapy regimen had to be considered unsuitable for second-line chemotherapy. RESULTS: The disease control rate was 32% (95% CI: 15.8-48.7) (1/31 patients had a partial response and 9/31 patients had stable disease) and the median overall survival 23 weeks (range 4 40). Symptom improvement was reported by 39% of patients overall and by 83% of patients who achieved disease control. The median time to symptom improvement was 3 weeks (range 2-4). Adverse events were generally mild (grade I or II) and reversible and consisted mostly of skin rash, diarrhea and fatigue. CONCLUSIONS: ZD1839 demonstrated clinically meaningful antitumor activity with significant improvement in symptoms in this heavily pretreated group of patients with advanced NSCLC. Furthermore, ZD1839 showed a favorable toxicity profile, with the majority of adverse events being mild and reversible. PMID- 12781430 TI - Stereotactic body frame based fractionated radiosurgery on consecutive days for primary or metastatic tumors in the lung. AB - To evaluate the feasibility and treatment outcomes of stereotactic radiosurgery (SRS) using a stereotactic body frame (Precision Therapy), we prospectively reviewed 34 tumors of the 28 patients with primary or metastatic intrathoracic lung tumors. Eligible patients included were nine with primary lung cancer and 19 with metastatic tumors from the lung, liver, and many other organs. A single dose of 10 Gy to the clinical target volume (CTV) was delivered to a total dose of 30 40 Gy with three to four fractions. Four to eight coplanar or non-coplanar static fields were generated to adequately cover the planning target volume (PTV) as well as to exclude the critical structures as much as possible. More than 90% of the PTV was delivered the prescribed dose in the majority of cases (average; 96%, range; 74-100%). The mean PTV was 41.4 cm(3) ranging from 4.4 to 230 cm(3). Set up error was within 5 mm in all directions (X, Y, Z axis). The response was evaluated by using a chest CT and/or 18FDG-PET scans after SRS treatment, 11 patients (39%) showed complete response, 12 (43%) partial response (decrease of more than 50% of the tumor volume), and four patients showed minimally decreased tumor volume or stable disease, but one patient showed progression disease. With a median follow-up period of 18 months, a local disease progression free interval was ranging from 7 to 35 months. Although all patients developed grade one radiation pneumonitis within 3 months, none had symptomatic or serious late complications after completing SRS treatment. Given these observations, it is concluded that the stereotactic body frame based SRS is a safe and effective treatment modality for the local management of primary or metastatic lung tumors. However, the optimum total dose and fractionation schedule used should be determined after the longer follow-up of these results. PMID- 12781431 TI - Impact of treatment interruptions due to toxicity on outcome of patients with early stage (I/II) non-small-cell lung cancer (NSCLC) treated with hyperfractionated radiation therapy alone. AB - We investigated the effect of treatment interruptions due to high-grade (> or =3) toxicity on outcome of patients with early stage (I/II) non-small-cell lung cancer treated with hyperfractionated radiation therapy (Hfx RT). Of 116 patients treated with total tumour doses of 69.6 Gy, 1.2 Gy b.i.d. fractionation, 44 patients refused surgery while 72 patients were medically inoperable due to existing co-morbid states. Patients who were medically inoperable had worse KPS (P=0.0059) and more pronounced weight loss (P=0.0005). Among them, 12 patients experienced high-grade toxicity and 11 of them with either acute (n=6) or "consequential" late (n=5) high-grade toxicity requested interruption in the Hfx RT course (range, 12-25 days; median, 17 days). Superior survival (OS) was observed in patients who refused surgery when compared to those who were medically inoperable (P=0.0041), as well as superior local recurrence-free survival (LRFS) (P=0.011), but not different distant metastasis-free survival (P=0.14). Cause-specific survival (CSS) also favoured patients who refused surgery (P=0.004). Multivariate analysis showed independent influence of the reason for not undergoing surgery on OS (P=0.035), but not on LRFS (P=0.084) or CSS (P=0.068). Patients who refused surgery did not experience high-grade toxicity (0/44), whereas 11 of 72 patients with medical inoperability and co morbid states experienced high-grade toxicity and had treatment interruptions to manage toxicity (P=0.0064). Patients without treatment interruptions had significantly better OS (P=0.00000), LRFS (P=0.00000) and CSS (P=0.00000) than those with treatment interruptions. When corrected for treatment interruptions, the reason for not undergoing surgery independently influenced OS (P=0.040), but not LRFS (P=0.092) or CSS (P=0.068). In contrast to this, treatment interruption was independent prognosticator of all three endpoints used (P=0.00031, P=0.0075 and P=0.00033, respectively). When 11 patients with treatment interruptions were excluded, the reason for not undergoing surgery still affected OS (P=0.037) and CSS (P=0.039) but not LRFS (P=0.11). Multivariate analyses using OS, CSS and LRFS showed that the reason for not undergoing surgery affected OS (P=0.0436), but neither CSS (P=0.083) nor LRFS (P=0.080). PMID- 12781432 TI - In vitro effects of combinations of cis-amminedichloro (2-methylpyridine) platinum (II) (ZD0473) with other novel anticancer drugs on the growth of SBC-3, a human small cell lung cancer cell line. AB - Among numerous clinical regimens of combination chemotherapy, synergy has been observed to be particularly marked with combinations containing cisplatin (CDDP). However, the clinical use of CDDP has sometimes been limited by acquired resistance. The new-generation platinum drug, ZD0473, was synthesized with the aim of hindering the reaction of the drug with thiols, by the introduction of a 2 methylpyridine ligand. This enables the drug to exert antitumor activity against cisplatin-resistant cancer cells with elevated glutathione and/or metallothionein levels. The drug was also shown experimentally to overcome cisplatin resistance due to impaired drug accumulation, and enhanced DNA repair/tolerance to platinum DNA adducts. We investigated the effects of combinations of ZD0473 with other anticancer drugs on the growth of a human small-cell lung cancer cell line (SBC 3). Six novel anticancer drugs were tested: docetaxel (TXT), paclitaxel (TXL), vinorelbine (VNB), irinotecan (CPT-11), gemcitabine (GEM) and pemetrexed (MTA). The growth inhibitory effect of the drugs was measured by MTT assay and the effects of the combination regimens were evaluated by the combination index analysis method developed by Chou and Talalay. Synergy was demonstrated for the combination regimens of ZD0473-GEM and ZD0473-TXL, while an additive effect was observed with combinations containing TXT, VNB, CPT-11 or MTA. In the case of the ZD0473-GEM combination, synergy was observed over a wide range of inhibition levels at dose ratios of 50:1, 100:1 and 250:1. The level of synergy was equivalent to that observed for combinations of CDDP-etoposide, CDDP-GEM and nedaplatin-CPT-11. The results suggest that the combination of ZD0473 with GEM merits further investigation in small cell lung cancer. PMID- 12781433 TI - Phase II study of irinotecan and carboplatin in patients with the refractory or relapsed small cell lung cancer. AB - We examined the safety and efficacy of the combination of irinotecan plus carboplatin in patients with refractory or relapsed small cell lung cancer (SCLC). Patients with previously treated SCLC were eligible. Patients were treated every 3 weeks with carboplatin (with a target area under the concentration versus time curve of 5 mg min/ml using the Calvert formula on day 1) plus irinotecan (50 mg/m(2) on days 1 and 8). From May 2000 to January 2002, 24 patients were eligible. None of the 22 patients achieved a complete response, but 15 achieved a partial response with an overall response rate of 68.2% (95% confidence interval, 45.1-86.1%). In 13 patients with sensitive disease, the response rate was 92.3% (95% confidence interval, 64.0-99.8%). The median survival time (MST) was 194 days (range 27-605 days). The MST did not differ significantly between patients with sensitive disease (245 days) and those with refractory disease (194 days, P=0.88). One patient died of treatment-related sepsis. Grade 3-4 hematologic toxicities included leukopenia in 58% of patients, neutropenia in 63%, thrombocytopenia in 58%, and anemia in 67%. Grade 3 diarrhea developed in 21% of patients and grade 3-4 infection in 13%. No patients had grade 4 diarrhea or grade 3-4 nausea and vomiting. This regimen is effective and well tolerated in patients with relapsed or refractory SCLC. However, the search for even more active regimens should be continued. PMID- 12781434 TI - Diffuse alveolar damage after ZD1839 therapy in a patient with non-small cell lung cancer. AB - ZD1839 is an orally active inhibitor selective for the epidermal growth factor receptor tyrosine kinase and has shown promise in the treatment of non-small cell lung cancer (NSCLC). We now present a case of diffuse alveolar damage (DAD) that developed in a 67-year-old man treated with ZD1839. On day 8 of ZD1839 administration, the patient complained of dyspnea and a new-ground glass opacity was apparent on a chest X-ray and computed tomography scan. Despite high-dose steroid therapy, the patient died 13 days after the first administration of ZD1839. Postmortem analysis of lung tissue revealed a pattern of DAD. No evidence of infection or of other specific etiologies was apparent. This case is the first reported of respiratory failure after ZD1839 treatment in a patient with NSCLC. Physicians should therefore be aware of the potential pulmonary toxicity of ZD1839. PMID- 12781435 TI - The role of house dust mites and other aeroallergens in atopic dermatitis. PMID- 12781436 TI - The evaluation and management of food allergy in atopic dermatitis. PMID- 12781437 TI - The use of corticosteroids and corticosteroid phobia in atopic dermatitis. PMID- 12781438 TI - Corticosteroid addiction and withdrawal in the atopic: the red burning skin syndrome. PMID- 12781439 TI - The treatment of atopic dermatitis with topical immunomodulators. PMID- 12781440 TI - The treatment of atopic dermatitis with systemic immunosuppressive agents. PMID- 12781441 TI - Phototherapy of atopic dermatitis. PMID- 12781442 TI - Five new cases of juvenile renal cell carcinoma with translocations involving Xp11.2: a cytogenetic and morphologic study. AB - Two cases of renal cell carcinoma (RCC) carrying a t(X;1)(p11.2;q21) in a 12-year old boy and a 14 year-old girl, two cases with a t(X;1)(p11.2;p34) in a 9-year old boy and a 31-year-old woman, and one case with a t(X;17)(p11.2;q25) in a 15 year-old boy are reported. Two are likely papillary RCC, with clear or slightly eosinophilic cells, and two to a clear cell RCC; one shows a mixture of papillary and clear cell RCC architecture. Renal cell carcinomas with translocations involving Xp11.2 form a specific entity characterized by subtle pathologic features and younger age of occurrence, especially for those with the t(X;17). PMID- 12781443 TI - Transfer of chromosome 8 into two breast cancer cell lines: total exclusion of three regions indicates location of putative in vitro growth suppressor genes. AB - Loss of heterozygosity (LOH) of the short arm of chromosome 8 occurs frequently in breast tumors. Fine mapping of the smallest regions of overlap of the deletions indicates that multiple tumor suppressor genes may be located in this region. We have performed microcell-mediated chromosome transfer of chromosome 8 into two breast cancer cell lines, 21MT-1 and T-47D. Twenty-two of the resulting hybrids were characterized extensively with chromosome 8 microsatellite markers and a subset were assayed for growth in vitro and soft agar clonicity. There was no evidence in any of the hybrids for suppression of growth or clonicity that could be attributed to the presence of particular regions of chromosome 8; however, none of the 22 hybrids examined had taken up all of the donor chromosome 8, and in fact there were three regions that contained only one allele of the markers genotyped in all 22 hybrids. These results are consistent with the presence of suppressor genes on the short arm of chromosome 8 causing strong growth suppression that is incompatible with growth in vitro; that is, multiple suppressor genes may exist on the short arm of chromosome 8. PMID- 12781444 TI - The effect of chlorambucil treatment on cytogenetic parameters in chronic lymphocytic leukemia patients. AB - The most common treatment of chronic lymphocytic leukemia (CLL) is the alkylating agent chlorambucil (CLB), with or without prednisone. In the present study, our aim was to evaluate whether treatment with CLB for more than one year induced genetic changes manifested by comparative genomic hybridization (CGH) as new chromosomal aberrations. We also studied whether CLB affected the pattern of replication by using fluorescence in situ hybridization (FISH). We found a similar rate of asynchronous pattern of replication in both treated and untreated patients with CLL. Most of the aberrations found with CGH were previously reported in CLL. More prognostically unfavorable aberrations and more cases with genetic changes were found in the treated group. The changes found were not typical of the secondary genetic aberrations associated with alkylating agents. Thus, we conclude that treatment of CLL with CLB for at least a year does not affect the parameters analyzed in this study. Longer studies are needed to further explore the effects of alkylating agents on normal and malignant cells. PMID- 12781445 TI - Evidence for involvement of a tumor suppressor gene on 1p in malignant peripheral nerve sheath tumors. AB - Malignant peripheral nerve sheath tumors (MPNST) are rare soft-tissue malignancies. The genetic basis of these tumors is still poorly understood. Cytogenetic analyses predominantly revealed complex karyotypes, precluding the identification of recurrent chromosomal changes. We report loss of 1p material in a near-diploid karyotype with few or no additional structural chromosome changes in two sporadic cases of MPNST, indicating an important role of 1p loss in MPNST development. In one of these two tumors, a distal 1p deletion (1p31.2 approximately pter) was detected suggesting involvement of a tumor suppressor gene located within this distal region of 1p. Further evidence for recurrent 1p loss in MPNST was obtained by interphase fluorescence in situ hybridization, which showed loss of 1p material in 3 out of 13 tumors. These findings together with data from the literature suggest that loss of a tumor suppressor gene located within distal 1p is implicated in the pathogenesis of MPNST. PMID- 12781446 TI - THY1 expression is associated with tumor suppression of human ovarian cancer. AB - Microcell-mediated transfer of chromosome 11 into the human ovarian cancer cell line SKOV-3 results in suppression of tumorigenicity in severe combined immunodeficiency (SCID) mice. To identify the differentially expressed transcripts associated with suppression of tumorigenicity, cDNA populations from the slow-growing tumorigenic clone 11(H)8-3, tumorigenic clone 11(H)8-4, and parental SKOV-3 cells were subtracted from the nontumorigenic clones, 11(H)7-2 and 11(C)9-8. The subtracted cDNA populations were either cloned, sequenced and searched in GenBank, or analyzed by gene discovery array screening. A cDNA transcript corresponding to the THY1 gene located at chromosome 11q23 approximately q24 was found to be exclusively expressed in the two nontumorigenic cell clones. In contrast, THY1 expression was not detected in SKOV-3, the tumorigenic hybrid clones, or six other tumorigenic ovarian cancer cell lines. Further analysis using immunocytochemistry and quantitative flow cytometry with a Thy-1-specific antibody confirmed the exclusive expression of THY1 at the protein level in the two nontumorigenic clones. Several cell growth and differentiation related genes, including thrombospondin 1 (THBS1), SPARC [secreted protein, acidic, cysteine-rich (osteonectin)], and fibronectin (FN1) were also found to be upregulated in the nontumorigenic clones; however, these were expressed in the slow-growing tumorigenic clones as well. Expression of these genes was not observed in the parental SKOV-3 cell line and therefore must be regulated by a gene or genes on chromosome 11. Our results suggest that THY1 is a putative tumor suppressor gene for ovarian cancer and that THBS1, SPARC, and FN1 are genes associated with the regulation of in vivo tumor growth rate. PMID- 12781447 TI - Modified allelic replication in lymphocytes of patients with neurofibromatosis type 1. AB - Transcription activity of genes is related to their replication timing, accordingly gene activation is coupled with a shift from late replication to early replication and vice versa. The relationship between replication timing and gene expression is best manifested by monoallelically expressed genes which show an asynchronous pattern of allelic replication, with the active allele replicating earlier than the inactive counterpart. Biallelically expressed genes, which normally replicate highly synchronously, when present in lymphocytes derived from patients with various types of malignancies or premalignancies, replicate highly asynchronously, similar to monoallelically expressed genes. Since neurofibromatosis-type 1 (NF1) patients are at an increased risk to develop malignancies, we used the fluorescence in situ hybridization (FISH) replication assay and evaluated the level of replication synchrony of three cancer-implicated genes (RB1, AML1, and CMYC) in lymphocytes derived from patients with NF1 without malignancy. Each gene, which normally displayed synchrony in allelic replication, in the patients' cells displayed loss of synchrony. The loss of replication synchrony, of each gene, in the patients' cells was achieved by an advanced replication of a single allele, which replicated remarkably earlier than its normal scheduled timing. In addition, the second allele showed slightly earlier replication timing than that normal for the gene. Thus, it is assumed that the NF1 condition is associated with activation of cancer-implicated genes that may be the cause for increased risk of patients to develop malignancies. As loss of synchrony in allelic replication timing differentiates well between NF1 patients and control subjects, this marker may have a potential use for identification of presymptomatic carriers of NF1 disorders. PMID- 12781448 TI - Amplification of the BCR/ABL fusion gene clustered on a masked Philadelphia chromosome in a patient with myeloblastic crisis of chronic myelocytic leukemia. AB - Although the chronic phase of chronic myelocytic leukemia (CML) is characterized by the Philadelphia (Ph) chromosome creating a hybrid BCR/ABL gene, additional genetic changes involved in blast crisis are poorly understood. We report a 4-8 fold amplification by tandem duplication of the BCR/ABL fusion gene clustered on a masked Ph chromosome in a 61-year-old male patient with CML in myeloblastic crisis. Our finding suggests that the BCR/ABL amplification may play a role as a novel mechanism in the progression to an aggressive blast transformation in some cases of Ph-positive CML. PMID- 12781449 TI - Expression of HIF-1 and ubiquitin in conventional renal cell carcinoma: relationship to mutations of the von Hippel-Lindau tumor suppressor gene. AB - Conventional clear cell renal cell carcinomas (cRCC) have mutations of the von Hippel-Lindau (VHL) tumor suppressor gene at 3p25 in approximately 50% of cases. The VHL gene normally regulates ubiquitin-mediated proteolysis of hypoxia inducible factor 1alpha (HIF-1alpha); in cell lines, VHL inactivation blocks HIF 1alpha proteolysis, resulting in increased HIF-1 expression. This study was undertaken to investigate the relationship between VHL mutations and the expression of ubiquitin and HIF-1alpha in cRCC. Eleven cRCC were studied with microsatellite analysis for 3p deletions and with sequencing for VHL mutations. Immunohistochemistry was performed for HIF-1alpha and ubiquitin. Deletions at 3p25 were found in 10 tumors, and VHL mutations were identified in 6 of these cases. There was staining for ubiquitin and HIF-1alpha in all tumors with VHL mutations. Among the five cases without VHL mutations, staining for ubiquitin or HIF-1alpha was not present in three cases but was present in two tumors, both of which had 3p deletions. The findings support a role for VHL mutations promoting cRCC development by an impairment of HIF-1alpha proteolysis. The findings also suggest that a 3p tumor suppressor gene other than VHL may also influence HIF 1alpha degradation and that there is an additional tumorigenic pathway for cRCC that does not involve VHL or HIF-1. PMID- 12781450 TI - Nodal marginal zone B-cell lymphoma with a novel t(X;5)(q28;q22): conventional and molecular cytogenetic analysis. AB - Cytogenetic studies provide important information for the diagnosis and classification of malignant lymphomas that in some cases also has prognostic significance. Furthermore, the investigation of isolated novel cytogenetic findings in malignant lymphoma has led to the discovery of many important oncogenes and tumor suppressor genes. For this reason, a case of nodal marginal zone B-cell lymphoma in a 72-year-old woman is described in which analysis by conventional and molecular cytogenetic techniques demonstrated the presence of a t(X:5)(q28;q22) as the sole chromosomal abnormality. This translocation has not been previously reported in the literature. PMID- 12781451 TI - HMGA2 is expressed in an allele-specific manner in human lipomas. AB - The architectural transcription factor HMGA2 is almost exclusively expressed in undifferentiated mesenchymal cells. Interestingly, it has been mapped to the translocation site in a variety of human mesenchymal tumors that reveal a terminally differentiated phenotype. The expression of chimeric HMGA2 transcripts encoding three DNA-binding domains fused to novel transcriptional regulatory domains was previously described in lipomas. In this study with lipoma ST91-198, we report the expression of truncated HMGA2 transcripts that gained no functional domains. The highly polymorphic region in the 5' untranslated region (UTR) of HMGA2 was used to determine the allele-specific expression of HMGA2 in lipomas. Microsatellite PCR revealed a monoallelic expression pattern, and only the translocated allele was expressed when the DNA-binding domains of the rearranged allele were fused with transcription activation domains. Surprisingly, a diallelic expression pattern of HMGA2 was observed in lipoma ST91-198, and the wild-type allele was also expressed. In conjunction with studies involving rearrangements of HMGA genes in other benign mesenchymal tumors, our results support a model in which the expression of the wild-type HMGA allele is critical for the pathogenesis of mesenchymal tumors and in which rearrangements of HMGA do not lead to a gain of function in the chimeric HMGA protein. PMID- 12781452 TI - Chromosomal abnormalities in inflammatory pseudotumor of the urinary bladder. AB - Inflammatory pseudotumors of the urinary bladder are rare, benign, nonepithelial tumors. Fewer than 30 have been reported, and no data are available on their karyotypic characteristics and/or the molecular mechanisms of pathogenesis. We performed short-term culturing and cytogenetic analysis of an inflammatory pseudotumor of the bladder, finding a der(20)t(12;20)(q13 approximately q15;q13) as the only cytogenetic aberration. The detection of a 12q13 approximately q15 rearrangement in the inflammatory pseudotumor indicates that this lesion is pathogenetically related to other benign mesenchymal tumors displaying, for example, lipogenic or leiomyomatous differentiation, something that is in sharp contrast to the karyotypic profile of epithelial tumors of the urinary bladder mucosa. PMID- 12781453 TI - Cytogenetic findings in familial B-cell chronic lymphocytic leukemia: a report of two cases in a family. AB - Familial B-cell chronic lymphocytic (B-CLL) leukemia has been defined as an entity epidemiologically different from sporadic B-CLL. Cytogenetic abnormalities in familial B-CLL, studied either by conventional cytogenetics or by interphase fluorescence in situ hybridization (i-FISH), have rarely been reported. We report a two-case family affected with B-CLL showing two different abnormal karyotypes detected by conventional cytogenetics [46,XX,del(7)(q32) and 46,XY,add(1)(p36),del(6)(q21)] but sharing a del(13)(q14) at the D13S319 locus, detected by interphase fluorescence in situ hybridization. PMID- 12781454 TI - Gonadoblastoma in a patient with del(9)(p22) and sex reversal: report of a case and review of the literature. AB - Studies of distinct clinical prototypes have significantly contributed to our understanding of evolutionary abnormalities and their association with neoplasia. We describe a phenotypic female, aged 20 years at report, who was examined as an infant for developmental retardation. The clinical characteristics of the 9p- syndrome were present and the external genitalia were those of a normal female. The karyotype was 46XY,del(9)(p22). The parental karyotypes were normal. No SRY deletion or mutation was detected. Sonography showed the presence of a uterus. Basal luteinizing hormone values were normal; follicle stimulating hormone values were high (40 IU/L). Stimulation with human chorionic gonadotropin did not produce any rise in testosterone. The gonads were removed and histologic analysis disclosed dysgenetic gonads with gonadoblastoma in situ. This case constitutes the fourth case of gonadoblastoma developing in an individual with 9p- syndrome and sex reversal. This and analogous prototypes point to a locus (or loci) on the short arm of chromosome 9, which either constitutes a nonspecific suppressor gene or a gonadoblastoma suppressor gene. An alternative hypothesis would be that a gonad not normally differentiated is more prone to gonadoblastoma development without any specific gene involvement. PMID- 12781455 TI - Secondary myelodysplastic syndrome after treatment for promyelocytic leukemia: clinical and genetic features of two cases. AB - Acute promyelocytic leukemia (APL) represents a biologic and clinically well defined subtype of acute nonlymphocytic leukemia with specific morphologic and karyotypic characteristics. Although secondary leukemia and myelodysplastic syndromes (MDS) are the most frequent secondary neoplasms following chemotherapy for acute leukemia, their development after complete remission in patients with APL is uncommon. We describe the clinical and genetic features of two APL patients who achieved CR after chemotherapy and all-trans retinoid acid treatment and subsequently developed a MDS. Therapy-related MDS karyotype changes such as abnormalities of chromosomes 5 and 7 were found in the cytogenetic analysis. Since TP53 alteration was detected in one case, possible implications of these findings in the onset of MDS are discussed. PMID- 12781456 TI - Fourier transform ion cyclotron resonance mass spectrometry with NanoLC/microelectrospray ionization and matrix-assisted laser desorption/ionization: analytical performance in peptide mass fingerprint analysis. AB - Protein identifications by peptide mass fingerprint analyses with Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) were performed using microelectrospray ionization coupled to nano liquid chromatography (NanoLC), as well as using matrix-assisted laser desorption/ionization (MALDI). Tryptic digests of bovine serum albumin (BSA), diluted down to femtomole quantities, have been desalted by fast NanoLC under isocratic elution conditions as the high resolving power of FT-ICR MS enables peptides to be separated during the mass analysis stage of the experiment. The high mass accuracy achieved with FT-ICR MS (a few ppm with external calibration) facilitated unambiguous protein identification from protein database searches, even when only a few tryptic peptides of a protein were detected. Statistical confidence in the database search results was further improved by internal calibration due to increased mass accuracy. Matrix-assisted laser desorption/ionization and micro electrospray ionization (ESI) FT-ICR showed good mass accuracies in the low femtomole range, yet a better sensitivity was observed with MALDI. However, in higher femtomole ranges slightly lower mass accuracies were observed with MALDI FT-ICR than with microESI FT-ICR due to scan-to-scan variations of the ion population in the ICR cell. Database search results and protein sequence coverage results from NanoLC FT-ICR MS and MALDI FT-ICR MS, as well as the effect of mass accuracy on protein identification for the peptide mass fingerprint analysis are evaluated. PMID- 12781457 TI - Direct determination of polycyclic aromatic hydrocarbons in solid matrices using laser desorption/laser photoionization ion trap mass spectrometry. AB - The development and characterization of a new instrument for solid sampling which couples IR laser desorption followed by UV laser photo-ionization and analysis using an ion trap mass spectrometer has been investigated. For calibration, a new type of solid sample preparation involving activated charcoal as the solid substrate was used. This solid sample provided a steady signal for several thousand laser shots, which allowed optimization of the experimental procedure. It was found that both the IR and UV intensity and the delay between them play an important role in both the magnitude and type of signals observed. A method of gas phase accumulation with multiple laser shots was examined. Finally, this technique was demonstrated to be effective in providing direct qualitative information for N.I.S.T. SRM 1944 river sediment sample with no sample pre treatment. PMID- 12781458 TI - Evaluation of chiral recognition characteristics of metal and proton complexes of di-o-benzoyl-tartaric acid dibutyl ester and L-tryptophan in the gas phase. AB - Chiral recognition of di-o-benzoyl-tartaric acid dibutyl ester (T) was achieved in the gas phase by electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry. In this method two divalent transition metal cations, zinc(II) and copper(II), were used as binding metal ions, and L tryptophan (A) was used as a chiral reference. Multimeric complexes were readily formed by electrospray ionization of a methanol:water (50:50) solution containing metal ion, L-tryptophan and T. These multimeric complexes included singly charged protonated dimeric [TAH](+), doubly charged copper(II) bound tetrameric [TACu H](2)(2+) and doubly charged zinc(II) bound tetrameric [TAZn-H](2)(2+), together with other complexes. The mass-selected complex, i.e., [TAH](+), [TACu-H](2)(2+) and [TAZn-H](2)(2+), was used to acquire the second stage mass spectra. The chiral recognition capability of these three complexes was evaluated using the abundance ratios of daughter ion to parent ion. A high degree of chiral recognition ability was observed in [TACu-H](2)(2+) and [TAZn-H](2)(2+). It was found that the type of binding ion played an important role in the chiral recognition. Different binding ions exhibited distinctive dissociation pathways and unique chiral recognition characteristics. The present method is based not only on whole-molecule loss but also on fractional-molecule loss. In addition, the reproducibility of the chiral recognition method was confirmed by several determinations of the abundance ratios of daughter ion to parent ion with a fixed activation energy and with five different activation energies. It was also shown that this chiral recognition method can tolerate acid interference. PMID- 12781459 TI - Fragmentation of cationized phosphotyrosine containing peptides by atmospheric pressure MALDI/Ion trap mass spectrometry. AB - An investigation of phosphate loss from sodium-cationized phosphotyrosine containing peptide ions was conducted using liquid infrared (2.94 microm) atmospheric pressure matrix-assisted laser desorption/ionization (AP MALDI) coupled to an ion trap mass spectrometer (ITMS). Previous experiments in our laboratory explored the fragmentation patterns of protonated phosphotyrosine containing peptides, which experience a loss of 98 Da under CID conditions in the ITMS. This loss of 98 Da is unexpected for phosphotyrosine, given the structure of its side chain. Phosphate loss from phosphotyrosine residues seems to be dependent on the presence of arginine or lysine residues in the peptide sequence. In the absence of a basic residue, the protonated phosphotyrosine peptides do not undergo losses of HPO(3) (Delta 80 Da) nor HPO(3) + H(2)O (Delta 98 Da) in their CID spectra. However, sodium cationized phosphotyrosine containing peptides that do not contain arginine or lysine residues within their sequences do undergo losses of HPO(3) (Delta 80 Da) and HPO(3) + H(2)O (Delta 98 Da) in their CID spectra. PMID- 12781460 TI - Loss of selenium from selenoproteins: conversion of selenocysteine to dehydroalanine in vitro. AB - Characterization of reduced and alkylated rat selenoprotein P by mass spectrometry yielded selenopeptides from which one or more selenium atoms were missing. Predicted selenopeptide mass peaks were accompanied by peaks corresponding to the conversion of one or more selenocysteine residues to dehydroalanine(s). Experiments were carried out to determine whether this loss of selenium occurred in vitro. A selenopeptide was isolated that contained two selenocysteine residues that were both in selenide-sulfide linkages with cysteine residues. After the peptide had been reduced and alkylated, in addition to the predicted mass peak with both selenocysteine residues present, two mass peaks were detected at positions expected for conversion of one and two selenocysteine residues of this selenopeptide to dehydroalanine residues, which was confirmed by tandem mass spectrometry. Similar findings were obtained from a study of another selenoprotein, rat plasma glutathione peroxidase. These results indicate that selenium atoms are lost from selenoproteins during purification and characterization. The loss of selenium from selenoproteins is probably through the mechanism of oxidation of selenocysteine residue to selenoxide followed by syn-beta-elimination of selenenic acid during sample processing. PMID- 12781461 TI - Advantages of Thermococcus kodakaraenis (KOD) DNA Polymerase for PCR-mass spectrometry based analyses. AB - The advantages of the thermostable DNA polymerase from Thermococcus kodakaraensis (KOD) are demonstrated for PCR amplification with subsequent detection by mass spectrometry. Commonly used DNA polymerases for PCR amplification include those from Thermus aquaticus (Taq) and Pyrococcus furiosus (Pfu). A 116 base-pair PCR product derived from a vWA locus was amplified by Taq, Pfu, or KOD DNA polymerase and compared by agarose gel electrophoresis and electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (ESI-FTICR-MS). KOD DNA polymerase demonstrated a 2- to 3-fold increase in PCR product formation compared to Pfu or Taq, respectively, and generated blunt-ended PCR product which allows facile interpretation of the mass spectrum. Additionally, we demonstrate the advantage of using high magnetic fields to obtain unit resolution of the same 116 base pair (approximately 72 kDa) PCR product at high m/z. PMID- 12781463 TI - Photodissociation of small group-11 metal cluster ions: fragmentation pathways and photoabsorption cross sections. AB - Noble metal cluster ions Cu(n)(+), Ag(n)(+) and Au(n)(+) (n = 3-21) have been stored in a Penning trap and photodissociated by low intensity laser pulses of 10 ns at photon energies of 3.49 eV and 4.66 eV. The fragmentation pathways, neutral monomer and dimer evaporation, have been monitored as a function of cluster size, excitation energy and element. It is found that the behavior of the branching ratio between monomer and dimer evaporation as a function of excitation energy depends on the metal under investigation. In particular, the slope of the energy dependence is positive for silver but negative for gold and copper cluster ions. Furthermore, photoabsorption cross sections are determined from observed total fragment yields in single-photon dissociation. PMID- 12781462 TI - Detection and characterization of methionine oxidation in peptides by collision induced dissociation and electron capture dissociation. AB - Electron capture dissociation (ECD) and collision-induced dissociation (CID), the two complementary fragmentation techniques, are demonstrated to be effective in the detection and localization of the methionine sulfoxide [Met(O)] residues in peptides using Fourier transform ion cyclotron resonance (FTICR) mass spectrometry. The presence of Met(O) can be easily recognized in the low-energy CID spectrum showing the characteristic loss of methanesulfenic acid (CH(3)SOH, 64 Da) from the side chain of Met(O). The position of Met(O) can then be localized by ECD which is capable of providing extensive peptide backbone fragmentation without detaching the labile Met(O) side chain. We studied CID and ECD of several Met(O)-containing peptides that included the 44-residue human growth hormone-releasing factor (GRF) and the human atrial natriuretic peptide (ANP). The distinction and complementarity of the two fragmentation techniques were particularly remarkable in their effects on ANP, a disulfide bond-containing peptide. While the predominant fragmentation pathway in CID of ANP was the loss of CH(3)SOH (64 Da) from the molecular ion, ECD of ANP resulted in many sequence informative products, including those from cleavages within the disulfide-bonded cyclic structure, to allow for the direct localization of Met(O) without the typical procedures for disulfide bond reduction followed by [bond]SH alkylation. PMID- 12781464 TI - Resonant excitation in a low-pressure linear ion trap. AB - It has been shown that through the process of resonant excitation the fragmentation of ions confined in a low-pressure (<0.05 mTorr) linear ion trap (LIT) can be accomplished while maintaining both high fragmentation efficiency and high resolution of excitation. The ion reserpine, 609.23 Da, has been fragmented with efficiencies greater than 90% while a higher mass ion, a homogeneously substituted triazatriphosphorine of mass 2721.89 Da, has been fragmented with 48% efficiency. This was accomplished by extended resonant excitation by low-amplitude auxiliary RF signals. Computer modelling of ion trajectories and analysis of the trapping potentials have demonstrated that a reduction in neutralization of ions on the rods (or losses on the rods) and increased fragmentation is a consequence of higher order terms in the potential introduced by the round-rod geometry of the LIT. PMID- 12781466 TI - Characterization and relative ionization efficiencies of end-functionalized polystyrenes by matrix-assisted laser desorption/ionization mass spectrometry. AB - The properties and relative ionization efficiencies of a series of polystyrenes (PS) with hydroxyl, hydrogen, tertiary amine, and quaternary amine end functionalities were examined by matrix-assisted laser desorption/ionization (MALDI) time-of-flight (TOF) mass spectrometry (MS). The hydrogen and hydroxyl functionalized PS ionized via attachment of a single Ag(+) cation, as expected. However, tertiary amine PS oligomers were found in (M - H)(+) and (M + H)(+) forms, in addition to M + Ag(+), while the quaternary amine PS oligomers only ionized to the M(+) form. Analysis of tertiary amine PS spectra revealed varying ratios of the three ionic forms depending on the oligomer length, pointing to a change in ionization efficiency. When the bulk samples were compared, the quaternary amine end-functionalized PS gave a ten-fold higher ionization efficiency over all others studied, likely because of the preexisting charge on the functionality. Samples with hydroxyl and hydrogen functionalities had similar ionization efficiencies, with the tertiary amine slightly higher, depending on the molecular weight. Changes in molecular weight affected the relative ionization efficiencies in varying fashion depending on the end functionality, though average molecular weight measurements were largely unaffected by end functionality. Quantification of end-functionalized polystyrenes with different ionization efficiencies was found to be possible if due care was taken. PMID- 12781465 TI - Mass spectrometric characterization of transferrins and their fragments derived by reduction of disulfide bonds. AB - Mass spectrometry, proteomics, and protein chemistry methods are used to characterize the cleavage products of 79 kDa transferrin proteins induced by iron catalyzed oxidation, including a novel C-terminal polypeptide released upon disulfide reduction. Top-down electrospray ionization tandem mass spectrometry (ESI-MS/MS) of intact multiply-charged transferrin from a variety of species (human, bovine, rabbit, chicken) performed on a quadrupole time-of-flight mass spectrometer yields multiply-charged b(n)-products originating near residues 56 69 from the N-terminal region, in addition to their complementary y(n)-products. Incubation of transferrin with reductants, such as dithiothreitol (DTT) or tris(2 carboxyethyl)-phosphine (TCEP), yields an increase in multiple charging observed by ESI-MS and an increase in molecular weight consistent with disulfide reduction. However, mammalian transferrins release a 6-8 kDa fragment upon disulfide reduction. Protein acetylation and MS/MS sequencing demonstrate that the fragment originates from the C-terminus of the protein, and that it is a separate polypeptide linked via three disulfide bonds to the main transferrin chain. The existence of a separate C-terminal chain is not annotated in protein sequence databases and, to date, has not been reported in the literature. Iron catalyzed cleavage induces fragments originating from both the N- and C-terminus of transferrin. PMID- 12781467 TI - Effect of the location of hydrogen abstraction on the fragmentation of diuretics in negative electrospray ionization mass spectrometry. AB - The diuretic agents bumetanide, xipamide, indapamide, and related compounds were investigated in order to determine the effect of different ionization sites on their collisionally activated dissociation and the corresponding fragmentation pathways. Therefore, analytes were selectively alkylated, and structural analogues as well as deuterium labeled compounds synthesized, which contain a reduced number of ionizable hydrogen atoms. Thus, specific hydrogen abstractions and their correlated dissociation routes of the negatively charged molecules were eliminated, providing evidence for the influence of the location of ionization on product ion spectra. Fragment ions such as m/z 78 indicate ionization at the commonly present sulfamoyl residue of diuretics but does not exclude additional ionization sites. Product ion spectra of the investigated diuretic agents proved to be composed by fragmentations initiated from different hydrogen abstractions. Moreover, the generation of radical anions by collision-activated dissociation of even-electron precursor ions was observed, the generation of which is discussed by proposed fragmentation pathways. PMID- 12781468 TI - Tandem mass spectrometry of metal nitrate negative ions produced by electrospray ionization. AB - M(NO(3))(x)(-) ions are generated by electrospray ionization (ESI) of metal solutions in nitric acid in negative ion mode. Collision-induced reactions of these ions are monitored in a tandem mass spectrometer (MS) of quadrupole octopole-quadrupole (QoQ) geometry. For Group 1 and 2 elements, the M(NO(3))(x)( ) ions dissociate into NO(3)(-) and neutral metal nitrate molecules. These elements also form some M(x)(NO(3))x+1- clusters, especially Li(+). Metal nitrate ions from transition elements and Group 13 elements fragment into oxo products and also undergo internal electron transfer to leave the M atom in a lower oxidation state. To calibrate the collision energy, the dissociation energy of O NO(2)(-) is found to be 5.55 eV, about 0.76 eV lower than a value derived from thermochemistry. The product ions from Fe(NO(3))(4)(-) ions have low formation thresholds of only 0.5 to 2 eV. PMID- 12781470 TI - A field evaluation of PCR for the routine detection of Babesia equi in horses. AB - We report on a study that evaluated the usefulness of PCR for the routine detection of Babesia equi in horses. The blood from a total of 105 horses comprising both sick and apparently healthy animals were examined for the presence of B. equi using both Wright-Giemsa-stained blood smears and PCR. Microscopic analysis of Giemsa-stained blood smears revealed 10/105 animals positive for Babesia, compared to 16/105 for the primary PCR and 36/105 for the nested PCR. Three of the 10 samples positive by Wright-Giemsa-stain were negative by PCR for B. equi. However, evidence is presented that these samples contained B. caballi and not B. equi. The Wright-Giemsa-stain was shown to identify Babesia in mostly clinically ill animals while the nested PCR detected the organism in a large number of apparently healthy animals. The results of this study suggest that the nested PCR is superior to both Wright-Giemsa-stained and primary PCR methods, and should be considered for the routine detection of B. equi in both healthy and clinically ill horses. PMID- 12781471 TI - Isolation and molecular characterization of Toxoplasma gondii from chickens and ducks from Egypt. AB - The prevalence of Toxoplasma gondii in free range chickens is a good indicator of the prevalence of T. gondii oocysts in the environment because chickens feed from the ground. In the present study, prevalence of T. gondii in 121 free range chickens (Gallus domesticus) and 19 ducks (Anas sp.) from a rural area surrounding Giza, Egypt was assessed. Blood, heart, and brain from each animal were examined for T. gondii infection. Antibodies to T. gondii, assayed with the modified agglutination test (MAT), were found in 49 (40.4%) chickens in titers of 1:5 in 11, 1:10 in four, 1:20 in four, 1:40 in eight, 1:80 in 10, and 1:160 or more in 12 chickens. Antibodies were found in three ducks each with a titer of 1:80. Hearts and brains of seropositive (MAT > or = 1:5) chickens and ducks were bioassayed in mice. Additionally, hearts and brains of seronegative (MAT<1:5) animals were bioassayed in T. gondii-free cats. T. gondii was isolated from 19 of 49 seropositive chickens (one with a titer of 1:5, two with a titer of 1:20, one with a titer of 1:40, five with a titer of 1:80, three with a titer of 1:160, and seven with a titer of > or = 1:360). One cat fed tissues pooled from 15 seronegative chickens shed T. gondii oocysts, while two cats fed tissues of 34 seronegative chickens did not shed oocysts. T. gondii was isolated from one of the seropositive ducks by bioassay in mice. The two cats fed tissues from 16 seronegative ducks did not shed oocysts. Genotyping of 20 chicken isolates of T. gondii using the SAG 2 locus indicated that 17 isolates were type III and three were type II. The duck isolate of T. gondii was type III. The mice inoculated with tissue stages of all 21 isolates of T. gondii from chickens and ducks remained asymptomatic, indicating that phenotypically they were not type I because type I strains are lethal for mice. Infections with mixed genotypes were not found. PMID- 12781472 TI - A follow-up of Beagle dogs intradermally infected with Leishmania chagasi in the presence or absence of sand fly saliva. AB - In this study, we compare the development of infection and/or disease in Beagle dogs intradermally infected with Leishmania chagasi, in the presence or absence of Lutzomyia longipalpis saliva, with those of intravenously infected animals. Spleen samples of all the animals inoculated with parasites had positive polymerase chain reaction tests for Leishmania DNA. Positive spleen cultures for Leishmania were detected earlier (P < or = 0.018) and were more frequent (five out of the five animals) in intravenously infected animals than in the intradermally infected animals, in presence (two out of the six animals) or absence (three out of the five animals) of salivary gland lysate of L. longipalpis. Significant increase in serum antibodies against Leishmania was observed only in the intravenously infected group (P = 0.004). In addition, dogs with infection confirmed by isolation of amastigotes or detection of parasite DNA were, nevertheless, negative for anti-Leishmania antibodies up to 5 months or more after infection. Only animals of the intravenously infected group developed progressive decreases in hematocrit (Pearson r = -0.8076, P = -0.0026) and hemoglobin (Pearson r = -0.8403, P = 0.0012) during the infection period. No significant difference in the course of infection was observed between groups of intradermally infected animals. The data presented herein confirms that the intradermal inoculation of dogs with Leishmania produces an asymptomatic form of infection. It also fails to show an advantage in using L. longipalpis saliva as an infection-enhancing agent in experimental canine leishmaniasis. PMID- 12781473 TI - Prevalence and infection pattern of naturally acquired giardiasis and cryptosporidiosis in range beef calves and their dams. AB - The prevalence and infection pattern of naturally acquired giardiasis and cryptosporidiosis in 20 ranch raised beef calves and their dams from birth to weaning was determined. Rectal fecal samples were collected from calves at 3 days of age and weekly thereafter; cows' fecal samples were collected at the time of calving, 1 week later and four times during the summer grazing period. Blood samples were collected from the calves at 3 days of age to determine IgG(1) concentrations. Giardia lamblia cysts were shed by all 20 calves (100%) at some date during the duration of the study. However, only one calf (5%) shed Cryptosporidium parvum on two sample dates during the trial. Giardia cysts were first detected at 3.9+/-1.37 weeks of age with a range of 2-7 weeks of age. The geometric mean number of Giardia cysts in the calf feces increased from none at 1 week of age to a maximum of 2230 cysts/g of feces at 5 weeks of age and then decreased to 2 cysts/g at 25-27 weeks of age. Infection rate of calves shedding Giardia cysts peaked at 85% at 5 weeks of age and then decreased to 21% at 25-27 weeks of age. Giardia cysts, shed by calves peaked 1 week after initial shedding and decreased (P<0.05) for the remainder of the trial with the exception of week 3. There was a lower (P<0.05) percentage of calves shedding Giardia cysts weeks 3 10 and 15-25 compared to when shedding was first detected. All calves had complete or partial transfer of passive immunity as measured by IgG(1) levels. The rate of infection (15%) and the geometric mean number of Giardia cysts in the cows' feces (38.49 cysts/g) numerically increased at 1 week post-calving compared to levels at calving. The rate of infection (40%) numerically increased and the geometric mean number of Cryptosporidium andersoni oocysts in the cow feces (37.48 oocysts/g) increased (P<0.05) at 1 week post-calving and decreased to 0 at 13-16 weeks post-calving. This study is the first to document the cumulative prevalence and infection patterns of Giardia and Cryptosporidium in beef cattle under ranch conditions. PMID- 12781474 TI - Effect of a single dose of ponazuril on neural infection and clinical disease in Sarcocystis neurona-challenged interferon-gamma knockout mice. AB - Interferon gamma-knockout mice were challenged with 5000 Sarcocystis neurona sporocysts acquired from a naturally infected opossum. Ponazuril was administered once, by gavage, at day 1, 3, 7, 10, or 14 post-infection (pi). Ponazuril was given at either 20 or 200mg/kg. Mice that survived to day 30 pi were euthanized. Severity of CNS infection was quantified as schizont density in the cerebellum. Unchallenged mice in treatment and non-treatment groups remained free of disease and gained weight throughout the experiment. All challenged mice, regardless of treatment, developed histologic evidence of CNS infection even though clinical signs were prevented in some groups. The greatest treatment benefits were seen in mice given 200mg/kg ponazuril between days 4 and 14 pi. Weight gain over the course of the experiment occurred only in mice that were given 200mg/kg ponazuril on day 7 or 10 pi. With the exception of groups given 200mg/kg ponazuril on day 7 or 14 pi, mice in groups that got sporocysts developed abnormal neurologic signs. No deaths before day 30 pi occurred in mice given ponazuril at 20mg/kg on day 7 pi or 200mg/kg on day 1, 7, 10, or 14 pi. This effect was not significant. Mice given 200mg/kg on day 7 pi had significantly fewer cerebellar schizonts than did those of the control group that was not given ponazuril. These results indicate that single-dose administration of ponazuril for prevention of CNS infection is partially protective when given between days 4 and 14 pi. PMID- 12781475 TI - Evaluation of antigen and antibody rapid detection tests for Trypanosoma evansi infection in camels in Kenya. AB - The card agglutination test for Trypanosoma evansi (CATT/T. evansi) for the detection of antibodies, and Suratex for the detection of circulating antigens were compared in a cross-sectional study involving camels in eastern and central parts of Kenya. Of the 2227 camels screened, 2038 were owned by nomadic pastoralists in T. evansi endemic areas in eastern Kenya. A herd of 86 camels were from a ranch in Mugwoni. In Athi River area, 35 camels belonged to Kenya Trypanosomiasis Research Institute, and 68 were slaughter animals. Diagnostic sensitivity estimates were obtained by testing sera from 51 camels that had been found to be parasitologically positive by the haematocrit centrifugation technique, buffy-coat technique and mouse inoculation. Diagnostic specificity was estimated by testing sera from 35 camels known to be trypanosome-free. Positive and negative predictive values (NPVs) were calculated using a range of prevalence values. The sensitivity of CATT/T. evansi (68.6%) was higher than that of Suratex (58.8%), but not significantly. Both tests had equally high specificity (100%). The overall prevalence was 2.3% (51 out of 2227) by parasite detection, 32.2% (327 out of 1017) by CATT/T. evansi and 19.6% (188 out of 961) by Suratex. Overall, there was a positive association between CATT/T. evansi and Suratex though the strength of association was low (McNemar's test=46.12, P=0.001; kappa=0.26, CI: 0.20-0.33). Parasite prevalence ranged from 0% in several herds to 27.8% in a herd in Isiolo. Prevalence was highest in Isiolo with 2.5% (51 out of 2030) by parasitological detection, 38.8% (321 out of 828) by CATT/T. evansi and 21.9% (169 out of 772) by Suratex. In Mugwoni prevalence was 7 and 18% by CATT/T. evansi and Suratex, respectively, and no parasites were detected. In Athi River Suratex detected 2.9% (3 out of 103) positive while CATT/T. evansi and parasitological methods gave negative results. At prevalence values between 10 and 100%, CATT/T. evansi as well as Suratex had infinitely high positive predictive values, whereas Suratex had a lower NPV than CATT/T. evansi. In conclusion, results of this study showed that CATT/T. evansi and Suratex were able to detect aparasitaemic infections rapidly and were more sensitive than parasitological methods in revealing the true extent of trypanosomosis in a herd. The tests effectively complemented parasitological methods in the detection of T. evansi infections in camels. PMID- 12781476 TI - Modelling the transmission dynamics of Echinococcus granulosus in sheep and cattle in Kazakhstan. AB - Cystic echinococcosis, caused by Echinococcus granulosus, is an emerging disease in many parts of the world and, in particular, in eastern Europe and the former Soviet Union. This paper examines the abundance and prevalence of infection of E. granulosus in cattle and sheep in Kazakhstan. Observed data are fitted to a mathematical model in order to determine if the parasite population is partly regulated by intermediate host immunity and to define parameters in the model. Such data would be useful to develop simulation models for the control of this disease. Maximum likelihood techniques were used to define the parameters and their confidence limits in the model and the negative binomial distribution was used to define the error variance in the observed data. The results indicated that there are significant variations in the infection pressure to sheep depending on their location. In particular sheep from Almaty Oblast and from central and northern Kazakhstan appeared to have a greater exposure than sheep from Jambyl or South Kazakhstan Oblasts. The infection pressure to cattle was somewhat lower in comparison. In common with other similar studies, there was no evidence of parasite-induced immunity in sheep or cattle in Kazakhstan due to natural infection. The highest abundance and prevalence were seen in the oldest age classes of animals. PMID- 12781478 TI - Virulence-associated genes in Escherichia coli isolates from poultry with colibacillosis. AB - Avian pathogenic Escherichia coli, the causative agent of colibacillosis, harbors several putative virulence genes. In this study we examined by polymerase chain reaction (PCR) the presence of 16 of those genes in 200 colibacillosis isolates from our region. The seven virulence genes iutA, iss, cvaC, tsh, papC, papG and felA were detected significantly more often amongst colibacillosis isolates than in fecal isolates from healthy birds, thereby confirming their worldwide occurrence and possible pathogenic role in colibacillosis. However, several of those genes were not detected in many colibacillosis isolates, and none of them were detected in 27.5% of those isolates, which suggests that variants of those genes and yet undetected virulence factors should be searched for. PMID- 12781479 TI - Use of a macrophage cell line for rapid detection of Mycobacterium bovis in diagnostic samples. AB - Mycobacterium bovis isolation on bacteriological media from suspected cases of bovine tuberculosis (TB) demands laborious and time-consuming procedures. Even polymerase chain reaction (PCR) and radiometric analyses are secondary procedures and not alternatives to bacteriological procedures. Therefore, there is a need to develop new techniques aimed at rapid M. bovis detection in diagnostic samples. The human macrophage cell line THP-1 was thus investigated in experiments of M. bovis propagation and isolation from reference lymph node suspensions. THP-1 cells were shown to support a high-titered propagation within 48h of minute amounts of both M. bovis BCG and fully pathogenic M. bovis strain 503. A semi nested PCR for TB-complex-specific insertion sequence IS6110 revealed M. bovis infection in THP-1 cells. The same was true of a flow cytometry (FC) assay for expression of M. bovis chaperonin 10 in infected cells. The reduced time for isolation and identification of M. bovis (48-72h) and the consistency of the test results make the use of macrophage cell cultures attractive and cost-effective for veterinary laboratories involved in TB surveillance. PMID- 12781480 TI - Prevalence of cpb2, encoding beta2 toxin, in Clostridium perfringens field isolates: correlation of genotype with phenotype. AB - Beta2 toxin, encoded by the cpb2 gene, has been implicated in the pathogenesis of porcine, equine and bovine enteritis by type A Clostridium perfringens. By incorporating primers to cpb2 into a multiplex genotyping PCR, we screened 3270 field isolates of C. perfringens. Of these, 37.2% were PCR positive for the cpb2 gene. The majority of isolates from cases of porcine enteritis were positive for cpb2 (>85%), and this was even more true for C. perfringens isolated from cases of porcine neonatal enteritis (91.8%). In contrast, isolates from normal pigs only contained cpb2 in 11.1% of cases. The correlation between enteritis in other animal species and the presence of cpb2 was not so strong. cpb2 was found in 21.4% of C. perfringens isolates from cattle with enteritis, and in 47.3% of isolates from calves with enteritis or abomastitis. The prevalence of cpb2 varied with genotype, with type A isolates being positive for this gene in 35.1% of cases. Furthermore, enterotoxigenic type D or type E strains almost always carried cpb2. We cloned a 6xHIS-tagged beta2 (HIS-beta2) and used this protein to raise antiserum against beta2. Culture supernatants from 68 cpb2-positive and 13 cpb2-negative strains were tested for the presence of beta2 by Western blotting. In cpb2-positive isolates of porcine origin, beta2 was almost always detected (96.9%). However, in cpb2-positive isolates from other animal species, only 50.0% expressed beta2 protein. The high rate of cpb2-positivity among strains from neonatal pigs with enteritis and the high correlation of genotype with phenotype, supports the contention that beta2 toxin plays a role in the pathogenesis of these infections. However, it may be important to consider the use of an additional method for the detection of beta2 toxin in non-porcine cpb2-positive isolates when making claims about the role of beta2 in enteritis in non-porcine species. PMID- 12781481 TI - Binding of host glycosaminoglycans and milk proteins: possible role in the pathogenesis of Streptococcus uberis mastitis. AB - The role of indirect binding of host proteins through glycosaminoglycans (GAGs) on adherence and internalization of Streptococcus uberis to bovine mammary epithelial cells was evaluated. Preincubation of S. uberis with GAGs followed by incubation with fetal bovine serum (FBS), bovine milk or milk proteins resulted in greater adherence to and internalization of S. uberis into mammary epithelial cells than observed in untreated controls. Highest values were detected, when final incubation was done with milk. Greater adherence to and internalization into mammary epithelial cells were observed when heparin sulfate (HEP) and milk were used compared with any other GAG and FBS. When individual milk proteins were used, greatest adherence and internalization were observed when S. uberis strains were pretreated with HEP followed by treatment with beta-casein. The findings of this study illustrate a pathogenic strategy of S. uberis that may occur during the very early stages of infection. PMID- 12781482 TI - Partial genome sequencing of Rhodococcus equi ATCC 33701. AB - Preliminary analysis of a partial (30% coverage) genome sequence of Rhodococcus equi has revealed a number of important features. The most notable was the extent of the homology of genes identified with those of Mycobacterium tuberculosis. The similarities in the proportion of genes devoted to fatty acid degradation and to lipid biosynthesis was a striking but not surprising finding given the relatedness of these organisms and their success as intracellular pathogens. The rapid recent improvement in understanding of virulence in M. tuberculosis and other pathogenic mycobacteria has identified a large number of genes of putative or proven importance in virulence, homologs of many of which were also identified in R. equi. Although R. equi appears to have currently unique genes, and has important differences, its similarity to M. tuberculosis supports the need to understand the basis of virulence in this organism. The partial genome sequence will be a resource for workers interested in R. equi until such time as a full genome sequence has been characterized. PMID- 12781483 TI - Prevalence and identity of cdt-related sequences in necrotoxigenic Escherichia coli. AB - The cytolethal distending toxins (CDT) are responsible for the mitosis block at G2/M and the cycle arrest of cells in culture. Escherichia coli isolated from humans and animals with intestinal and extra-intestinal diseases can be positive for the production of a CDT-like cytopathic effect or for the presence of cdt related genes. The purpose of this study was to compare the prevalence and the identity of cdt-related sequences in necrotoxigenic E. coli (NTEC). A collection of 98 bovine type 2 NTEC (NTEC2) and 45 bovine, 20 canine, 3 feline, 65 human and 129 porcine type 1 NTEC (NTEC1) isolates was studied by colony hybridisation and PCR assays specific for the cdtB genes encoding the B sub-unit of the CDT-I, CDT II, CDT-III and CDT-IV toxins produced by E. coli. cdtB-III sequences were frequent amongst bovine NTEC2, since 83% of these isolates were positive by colony hybridisation and/or PCR, whereas cdtB-related sequences were rare amongst NTEC1, since only 2 bovine (4%), 3 canine (15%), 10 human (15%) and 13 porcine (10%) of these isolates were positive. The 28 probe-positive NTEC1 harboured cdtB IV sequences (13 isolates), cdtB-I sequences (10 isolates), or still unidentified cdt-related sequences (5 isolates). After comparison with previously published and unpublished results of phenotypic assay on cell cultures, existence of other cdt-related sequences is suggested amongst NTEC1. The differences between NTEC1 and NTEC2 in their CDT profiles may have implication for the pathogenesis of those two classes of pathogenic E. coli. PMID- 12781484 TI - Analysis of virulence plasmid gene expression of intra-macrophage and in vitro grown Rhodococcus equi ATCC 33701. AB - Rhodococcus equi is a soil organism that infects macrophages of foals and immunocompromised humans. Virulence in foal isolates is tightly associated with an 80kb plasmid, which includes a pathogenicity island (PI) with a virulence associated gene family, vap. A DNA microarray containing 66 of 69 putative open reading frames (ORFs) of the virulence plasmid was developed. Virulence plasmid gene expression of R. equi grown in macrophages or under different conditions in vitro was compared against in vitro growth at 30 degrees C, pH=7. When grown in macrophages, all seven vap family genes as well as six ORFs within, but not outside, the PI were induced. Cluster analysis of the gene expression matrix assembled from different growth conditions suggested that those genes that actively responded to environmental changes divided broadly into two groups. One group, orf1, 2, 5, 6-8, 12-15, 19, and 20 (which includes all the vap genes), was induced at 37 degrees C, mostly by low iron, and to a lesser extent by the synergy of low calcium and pH=5. The second group, orf3, 9, and 10, was induced at 37 degrees C by magnesium depletion (produced by EDTA treatment of growth medium). Temperature (37 degrees C) was the most important factor inducing gene expression for the both groups. Iron restriction led to down-regulation of Group II genes and magnesium restriction led to down-regulation of Group I genes. A putative consensus IdeR binding site was identified upstream of vapA, suggesting that vapA is a member of an IdeR regulon in R. equi. Expression of genes inside macrophages was most closely but not completely mimicked by growth of bacteria at 37 degrees C, pH=5, under conditions of restricted iron, calcium and magnesium; that is, similar to environmental factors found inside macrophages. PMID- 12781485 TI - A single agenda needed for malaria. PMID- 12781487 TI - Decontamination spray could save lives after bioterrorist attack. PMID- 12781490 TI - HIV mystery in the Philippines. PMID- 12781492 TI - Improved monitoring of febrile illnesses. PMID- 12781491 TI - Kenya rejects drugs deal. PMID- 12781493 TI - Malariotherapy to treat HIV patients? PMID- 12781494 TI - Antibacterial resistance deemed a public-health crisis. PMID- 12781497 TI - Worsening health situation in Basra. PMID- 12781498 TI - WHO is SARS? PMID- 12781500 TI - Anopheles gambiae and climate in Brazil. PMID- 12781501 TI - Was the Black Death yersinial plague? PMID- 12781503 TI - Severe acute respiratory syndrome in developing countries. PMID- 12781504 TI - Hyperlactataemia syndromes associated with HIV therapy. AB - Hyperlactataemia is seen in 8-18.3% of HIV-infected patients taking nucleoside analogue reverse transcriptase inhibitors (NRTIs). Recent epidemiological studies suggest that most episodes are transient and subclinical. However, symptomatic and occasionally life-threatening cases accompanied by metabolic acidosis and hepatic steatosis (ie, lactic acidosis syndrome) have also been described. Though yet to be fully elucidated, the proposed mechanism is NRTI-induced inhibition of mitochondrial DNA polymerase culminating in derangements in oxidative phosphorylation and lactate homeostasis. Signs and symptoms range from mild hyperlactataemia accompanied by nausea, abdominal discomfort, and weight loss to severe, intractable lactic acidosis complicated by coma and multi-organ failure. Significant progress has recently been made with regard to the natural history of NRTI-related hyperlactataemia. However, other important aspects of the disorder, such as its pathogenesis, predisposing conditions, and management, remain poorly understood. This article reviews the current published work on these issues, identifies areas of controversy, and addresses directions for future research. PMID- 12781505 TI - Marine natural products and their potential applications as anti-infective agents. AB - The oceans are a unique resource that provide a diverse array of natural products, primarily from invertebrates such as sponges, tunicates, bryozoans, and molluscs, and from marine bacteria and cyanobacteria. As infectious diseases evolve and develop resistance to existing pharmaceuticals, the marine environment provides novel leads against fungal, parasitic, bacterial, and viral diseases. Many marine natural products have successfully advanced to the late stages of clinical trials, including dolastatin 10, ecteinascidin-743, kahalalide F, and aplidine, and a growing number of candidates have been selected as promising leads for extended preclinical assessment. Although many marine-product clinical trials are for cancer chemotherapy, drug resistance, emerging infectious diseases, and the threat of bioterrorism have all contributed to the interest in assessing natural ocean products in the treatment of infectious organisms. In this review, we focus on the pharmacologically tested marine leads that have shown in-vivo efficacy or potent in-vitro activity against infectious and parasitic diseases. PMID- 12781506 TI - Rabbit's revenge. PMID- 12781507 TI - Measurement of trends in childhood malaria mortality in Africa: an assessment of progress toward targets based on verbal autopsy. AB - Reduction of deaths associated with malaria in children is a primary goal of malaria control programmes in Africa, but there has been little discussion about how changes in mortality will be measured. This paper assesses recent historical changes in the contribution of malaria to child survival in Africa by examining data from demographic surveillance systems (DSS) in 25 mainly rural settings. The data were adjusted for the varying sensitivity and specificity of verbal autopsies (VA) in different ranges of malaria mortality and for varying parasite prevalences. Average malaria mortality in the DSS sites in west Africa was 7.8 per 1000 child-years between 1982 and 1998; the rate did not change significantly over this period. In the sites in east and southern Africa combined, malaria mortality was 6.5 per 1000 child-years between 1982 and 1989, but it increased to 11.9 per 1000 child-years between 1990 and 1998. All-cause child mortality and non-malaria mortality, by contrast, decreased significantly over time in both regions; consequently, the proportion of deaths due to malaria rose from 18% to 23% in west African sites and from 18% to 37% in east and southern African sites between 1982-89 and 1990-98. If malaria mortality fell at a rate consistent with the Roll Back Malaria target of halving malaria mortality by the year 2010, an individual DSS of a total population of 63 500 could with adequate VA adjustment detect this reduction after 7 years. PMID- 12781509 TI - A critical review of the SAFE strategy for the prevention of blinding trachoma. AB - Trachoma is an ocular disease caused by repeated infection with Chlamydia trachomatis. It is the leading cause of infectious blindness globally, responsible for 5.9 million cases of blindness. Although trachomatous blindness is untreatable, it is eminently possible to prevent and the World Health Organization promotes the use of the SAFE strategy (surgery to treat end-stage disease, antibiotics to reduce the reservoir of infection, facial cleanliness, and environmental improvement to reduce transmission of C trachomatis) for this purpose. In this review we have assessed the evidence base supporting the elements of the SAFE strategy. We find strong support for the efficacy of the surgery and antibiotics components, although the optimal antibiotic regimens have not yet been established. The evidence for an effect of health education and environmental improvement is weaker, and depends mostly on cross-sectional observational studies. PMID- 12781508 TI - Immunomodulatory effects of quinolones. AB - We review data on the in-vitro, ex-vivo, in-vivo, and clinical effects of fluoroquinolones on the synthesis of cytokines and their mechanisms of immunomodulation. In general, most fluoroquinolone derivatives superinduce in vitro interleukin 2 synthesis but inhibit synthesis of interleukin 1 and tumour necrosis factor (TNF)alpha; furthermore, they enhance significantly the synthesis of colony-stimulating factors (CSF). Fluoroquinolones affect in-vivo cellular and humoral immunity by attenuating cytokine responses. Interleukins 10 and 12 have an important role in the functional differentiation of immunocompetent cells and trigger the initiation of the acquired immune response. In addition, certain fluoroquinolones were seen to enhance haematopoiesis by increasing the concentrations of CSF in the lung as well as in the bone marrow and shaft. Those fluoroquinolones exerting significant effects on haematopoiesis were those with a cyclopropyl moiety at position N1 of their quinolone core structure. Mechanisms that could explain the various immunomodulatory effects of fluoroquinolones include: (1) an effect on intracellular cyclic adenosine-3',5'-monophosphate and phosphodiesterases; (2) an effect on transcription factors such as nuclear factor (NF)kappaB, activator protein 1, NF-interleukin-6 and nuclear factor of activated T cells; and (3) a triggering effect on the eukaryotic equivalent of bacterial SOS response with its ensuing intracellular events. Further studies are required, especially in the clinical setting to exploit fully the potential of the immunomodulatory effect of fluoroquinolones during, for example, immunosuppression, chronic airway inflammatory diseases, and sinusitis. PMID- 12781511 TI - Ralph Steinman--pioneering new perspectives on the immune system and infectious diseases. Interviewed by Marilynn Larkin. PMID- 12781514 TI - Uncultivated organisms. PMID- 12781515 TI - Structural basis of the herpesvirus M3-chemokine interaction. AB - Viruses have been fighting the immune systems of their hosts for millions of years and have evolved evasion strategies to ensure their survival. Viruses can teach us efficient mechanisms to control the immune system, and this information can be used to design new strategies of immune modulation that we might apply to diminish immunopathological responses that cause human diseases. Large DNA viruses, such as poxviruses and herpesviruses, encode proteins that are secreted from infected cells, bind cytokines and neutralize their activity. A subgroup of these viral proteins binds chemokines, a complex family of cytokines that control the recruitment of cells to sites of infection and inflammation. One of the major unresolved questions in the field was to understand how these viral secreted proteins bind chemokines with high affinity, despite having no amino acid sequence similarity to the host chemokine receptors, which are seven transmembrane-domain proteins that cannot be engineered as soluble proteins. PMID- 12781516 TI - Is the bacterial ferrous iron transporter FeoB a living fossil? AB - The cytoplasmic membrane protein FeoB of Escherichia coli, Helicobacter pylori, Legionella pneumophila and Synechocystis sp. strain PCC 6803 is necessary for Fe(2+) uptake. The C-terminal part of FeoB is predicted to contain 8-12 membrane spanning helices. The N-terminal domain shows much similarity to eukaryotic and prokaryotic G proteins and, indeed, GTPase activity is necessary for Fe(2+) transport. Four of the five characteristic conserved G protein motifs have been identified in FeoB proteins. Whether FeoB is involved directly, via its Me(2+) binding site, or indirectly in Fe(2+) transport, remains to be investigated. PMID- 12781517 TI - Genomics of pyoverdine-mediated iron uptake in pseudomonads. AB - Pyoverdines (PVDs) are complex siderophores produced by members of the fluorescent Pseudomonas. They comprise a dihydroxyquinoline fluorescent chromophore joined to a peptide of remarkably variable length and composition. In Pseudomonas aeruginosa, PVDs also function as signal molecules for the production of virulence factors. Genes responsible for the biosynthesis, excretion, uptake and regulation of these high-affinity siderophores are located either at a single locus or at up to three different loci in the genomes of the four pseudomonads analyzed. The peptide backbone of PVD is assembled by non-ribosomal peptide synthetases (NRPSs) and modified by accessory enzymes in the cytoplasm, and probably the periplasm. Regulation of PVD production and uptake depends on two extracytoplasmic sigma factors (ECF-sigmas), PvdS and FpvI, together with one anti-sigma, FpvR. PMID- 12781518 TI - Molecular evolution of sensory domains in cyanobacterial chemoreceptors. AB - Components of the chemotaxis system encoded in multiple homologous operons were identified in five cyanobacterial genomes. Analysis of phylogenetic profiles, genomic context, domain architectures and sequence identity reveal that sensory modules of chemoreceptors that detect environmental cues are the subject of frequent domain birth and death events and have accelerated rates of sequence evolution. This fact could explain a remarkable diversity of the sensing repertoire of chemotaxis receptors in microorganisms. PMID- 12781519 TI - The good, the bad and the ugly? PMID- 12781520 TI - The virulence-transmission trade-off and virulence management. PMID- 12781523 TI - Neutrophil granulocytes--Trojan horses for Leishmania major and other intracellular microbes? AB - Polymorphonuclear neutrophil granulocytes (PMNs) possess numerous effector mechanisms to kill ingested pathogens as the first line of defence. However, several microorganisms evade intracellular killing in neutrophils, survive and retain infectivity. There is increasing evidence that several pathogens even multiply within neutrophils. Taking Leishmania major as a prototypic intracellular pathogen, we suggest an evasion strategy that includes the manipulation of PMNs in such a way that the pathogens are able to use the granulocytes as host cells. The ability to survive and maintain infectivity in PMNs subsequently enables these organisms to establish productive infection. These organisms can use granulocytes as Trojan horses before they enter their definitive host cells, the macrophages. PMID- 12781521 TI - Understanding and managing pathogen evolution: a way forward. PMID- 12781524 TI - What is the nature of the replicative niche of a stealthy bug named Brucella? AB - Brucella spp. are facultatively intracellular bacteria that persist and multiply in the macrophages of their mammalian hosts. The so-called phagosome to which they have adapted is their natural living niche. Characterization of this niche would facilitate an understanding of the true relationship between the host cell and the intracellular bacteria. This Opinion analyses and discusses the characteristic properties and genesis of this vacuole during phagocytosis as deduced from the virulence factors necessary for intracellular multiplication of the pathogen. We conclude that the replicative niche of Brucella spp.--the 'brucellosome'--differs from all other cellular organelles, and that it isolates the pathogen from certain cytoplasmic nutrients. Adaptation to the stress conditions encountered and the use of anaerobic respiration enable brucellae to replicate in the compartment they create. PMID- 12781525 TI - Life without dihydrofolate reductase FolA. AB - Reduced folate derivatives participate in numerous reactions of bacterial intermediary metabolism. Consequently, the well-characterized enzyme implicated in the formation of tetrahydrofolate--dihydrofolate reductase FolA--was considered to be essential for bacterial growth. However, comparative genomics has revealed several bacterial genome sequences that appear to lack the folA gene. Here, we provide in silico evidence indicating that folA-lacking bacteria use a recently discovered class of flavin-dependent thymidylate synthases for deoxythymidine-5'-monophosphate synthesis, and propose that many bacteria must contain uncharacterized sources for reduced folate molecules that are still waiting to be discovered. PMID- 12781526 TI - Virulence factor regulation and regulatory networks in Streptococcus pyogenes and their impact on pathogen-host interactions. AB - Streptococcus pyogenes (group A streptococcus, GAS) is a very important human pathogen with remarkable adaptation capabilities. Survival within the harsh host surroundings requires sensing potential on the bacterial side, which leads in particular to coordinately regulated virulence factor expression. GAS 'stand alone' response regulators (RRs) and two-component signal transduction systems (TCSs) link the signals from the host environment with adaptive responses of the bacterial cell. Numerous putative regulatory systems emerged from GAS genome sequences. Only three RRs [Mga, RofA-like protein (RALP) and Rgg/RopB] and three TCSs (CsrRS/CovRS, FasBCAX and Ihk/Irr) have been studied in some detail with respect to their growth-phase-dependent activity and their influence on GAS-host cell interaction. In particular, the Mga-, RALP- and Rgg/RopB-regulated pathways display interconnected activities that appear to influence GAS colonization, persistence and spreading mechanisms, in a growth-phase-related fashion. Here, we have summarized our current knowledge about these RRs and TCSs to highlight the questions that should be addressed in future research on GAS pathogenicity. PMID- 12781527 TI - Campylobacter--a tale of two protein glycosylation systems. AB - Post-translational glycosylation is a universal modification of proteins in eukarya, archaea and bacteria. Two recent publications describe the first confirmed report of a bacterial N-linked glycosylation pathway in the human gastrointestinal pathogen Campylobacter jejuni. In addition, an O-linked glycosylation pathway has been identified and characterized in C. jejuni and the related species Campylobacter coli. Both pathways have similarity to the respective N- and O-linked glycosylation processes in eukaryotes. In bacteria, homologues of the genes in both pathways are found in other organisms, the complex glycans linked to the glycoproteins share common biosynthetic precursors and these modifications could play similar biological roles. Thus, Campylobacter provides a unique model system for the elucidation and exploitation of glycoprotein biosynthesis. PMID- 12781528 TI - The coming crisis of long-term care. PMID- 12781529 TI - Comparative analysis of the SARS coronavirus genome: a good start to a long journey. PMID- 12781530 TI - Poststroke depression: getting the full picture. PMID- 12781531 TI - Using "rationally designed drugs" rationally. PMID- 12781532 TI - The 2003 NRPB report on UK nuclear-test veterans. PMID- 12781533 TI - Epidemiological determinants of spread of causal agent of severe acute respiratory syndrome in Hong Kong. AB - BACKGROUND: Health authorities worldwide, especially in the Asia Pacific region, are seeking effective public-health interventions in the continuing epidemic of severe acute respiratory syndrome (SARS). We assessed the epidemiology of SARS in Hong Kong. METHODS: We included 1425 cases reported up to April 28, 2003. An integrated database was constructed from several sources containing information on epidemiological, demographic, and clinical variables. We estimated the key epidemiological distributions: infection to onset, onset to admission, admission to death, and admission to discharge. We measured associations between the estimated case fatality rate and patients' age and the time from onset to admission. FINDINGS: After the initial phase of exponential growth, the rate of confirmed cases fell to less than 20 per day by April 28. Public-health interventions included encouragement to report to hospital rapidly after the onset of clinical symptoms, contact tracing for confirmed and suspected cases, and quarantining, monitoring, and restricting the travel of contacts. The mean incubation period of the disease is estimated to be 6.4 days (95% CI 5.2-7.7). The mean time from onset of clinical symptoms to admission to hospital varied between 3 and 5 days, with longer times earlier in the epidemic. The estimated case fatality rate was 13.2% (9.8-16.8) for patients younger than 60 years and 43.3% (35.2-52.4) for patients aged 60 years or older assuming a parametric gamma distribution. A non-parametric method yielded estimates of 6.8% (4.0-9.6) and 55.0% (45.3-64.7), respectively. Case clusters have played an important part in the course of the epidemic. INTERPRETATION: Patients' age was strongly associated with outcome. The time between onset of symptoms and admission to hospital did not alter outcome, but shorter intervals will be important to the wider population by restricting the infectious period before patients are placed in quarantine. PMID- 12781534 TI - Spinal AV malformation. PMID- 12781535 TI - Clinical progression and viral load in a community outbreak of coronavirus associated SARS pneumonia: a prospective study. AB - BACKGROUND: We investigated the temporal progression of the clinical, radiological, and virological changes in a community outbreak of severe acute respiratory syndrome (SARS). METHODS: We followed up 75 patients for 3 weeks managed with a standard treatment protocol of ribavirin and corticosteroids, and assessed the pattern of clinical disease, viral load, risk factors for poor clinical outcome, and the usefulness of virological diagnostic methods. FINDINGS: Fever and pneumonia initially improved but 64 (85%) patients developed recurrent fever after a mean of 8.9 (SD 3.1) days, 55 (73%) had watery diarrhoea after 7.5 (2.3) days, 60 (80%) had radiological worsening after 7.4 (2.2) days, and respiratory symptoms worsened in 34 (45%) after 8.6 (3.0) days. In 34 (45%) patients, improvement of initial pulmonary lesions was associated with appearance of new radiological lesions at other sites. Nine (12%) patients developed spontaneous pneumomediastinum and 15 (20%) developed acute respiratory distress syndrome (ARDS) in week 3. Quantitative reverse-transcriptase (RT) PCR of nasopharyngeal aspirates in 14 patients (four with ARDS) showed peak viral load at day 10, and at day 15 a load lower than at admission. Age and chronic hepatitis B virus infection treated with lamivudine were independent significant risk factors for progression to ARDS (p=0.001). SARS-associated coronavirus in faeces was seen on RT-PCR in 65 (97%) of 67 patients at day 14. The mean time to seroconversion was 20 days. INTERPRETATION: The consistent clinical progression, shifting radiological infiltrates, and an inverted V viral-load profile suggest that worsening in week 2 is unrelated to uncontrolled viral replication but may be related to immunopathological damage. PMID- 12781536 TI - Lung pathology of fatal severe acute respiratory syndrome. AB - BACKGROUND: Severe acute respiratory syndrome (SARS) is a novel infectious disease with global impact. A virus from the family Coronaviridae has been identified as the cause, but the pathogenesis is still unclear. METHODS: Post mortem tissue samples from six patients who died from SARS in February and March, 2003, and an open lung biopsy from one of these patients were studied by histology and virology. Only one full autopsy was done. Evidence of infection with the SARS-associated coronavirus (SARS-CoV) and human metapneumovirus was sought by reverse-transcriptase PCR and serology. Pathological samples were examined by light and electron microscopy and immunohistochemistry. FINDINGS: All six patients had serological evidence of recent infection with SARS-CoV. Diffuse alveolar damage was common but not universal. Morphological changes identified were bronchial epithelial denudation, loss of cilia, and squamous metaplasia. Secondary bacterial pneumonia was present in one case. A giant-cell infiltrate was seen in four patients, with a pronounced increase in macrophages in the alveoli and the interstitium of the lung. Haemophagocytosis was present in two patients. The alveolar pneumocytes also showed cytomegaly with granular amphophilic cytoplasm. The patient for whom full autopsy was done had atrophy of the white pulp of the spleen. Electron microscopy revealed viral particles in the cytoplasm of epithelial cells corresponding to coronavirus. INTERPRETATION: SARS is associated with epithelial-cell proliferation and an increase in macrophages in the lung. The presence of haemophagocytosis supports the contention that cytokine dysregulation may account, at least partly, for the severity of the clinical disease. The case definition of SARS should acknowledge the range of lung pathology associated with this disease. PMID- 12781537 TI - Comparative full-length genome sequence analysis of 14 SARS coronavirus isolates and common mutations associated with putative origins of infection. AB - BACKGROUND: The cause of severe acute respiratory syndrome (SARS) has been identified as a new coronavirus. Whole genome sequence analysis of various isolates might provide an indication of potential strain differences of this new virus. Moreover, mutation analysis will help to develop effective vaccines. METHODS: We sequenced the entire SARS viral genome of cultured isolates from the index case (SIN2500) presenting in Singapore, from three primary contacts (SIN2774, SIN2748, and SIN2677), and one secondary contact (SIN2679). These sequences were compared with the isolates from Canada (TOR2), Hong Kong (CUHK-W1 and HKU39849), Hanoi (URBANI), Guangzhou (GZ01), and Beijing (BJ01, BJ02, BJ03, BJ04). FINDINGS: We identified 129 sequence variations among the 14 isolates, with 16 recurrent variant sequences. Common variant sequences at four loci define two distinct genotypes of the SARS virus. One genotype was linked with infections originating in Hotel M in Hong Kong, the second contained isolates from Hong Kong, Guangzhou, and Beijing with no association with Hotel M (p<0.0001). Moreover, other common sequence variants further distinguished the geographical origins of the isolates, especially between Singapore and Beijing. INTERPRETATION: Despite the recent onset of the SARS epidemic, genetic signatures are emerging that partition the worldwide SARS viral isolates into groups on the basis of contact source history and geography. These signatures can be used to trace sources of infection. In addition, a common variant associated with a non conservative aminoacid change in the S1 region of the spike protein, suggests that immunological pressures might be starting to influence the evolution of the SARS virus in human populations. PMID- 12781538 TI - Asking the right question. PMID- 12781539 TI - Survival in patients with primary systemic amyloidosis and raised serum cardiac troponins. AB - Patients with primary systemic amyloidosis that affects the heart have a poor outlook. Cardiac troponins T and I (cTnT, cTnI) are highly specific and sensitive biomarkers of myocardial injury. Values of these troponins provide quantitative information about the disease. We retrospectively assessed 261 patients newly diagnosed as having primary systemic amyloidosis. Median survival for patients with detectable cTnT and cTnI (6 and 8 months, respectively), was worse than that for those with undetectable values (22 and 21 months, respectively). Median and 25th and 75th percentile values for cTnT were 0.024 microg/L, less than 0.01 microg/L, and 0.084 microg/L, and for cTnI were 0.1 microg/L, 0.05 microg/L, and 0.24 microg/L, respectively. After multivariate analysis, cTnT proved a better predictor of survival than cTnI. PMID- 12781540 TI - CRIB II: an update of the clinical risk index for babies score. AB - The clinical risk index for babies (CRIB) score is a risk-adjustment instrument widely used in neonatal intensive care. Its appropriateness with contemporary data has been questioned. We have examined these questions, developed a new five item CRIB II score with data from a UK-wide sample of infants admitted to neonatal intensive care in 1998-99, and shown how mortality after neonatal intensive care has fallen in the past 12 years. CRIB II provides a recalibrated and simplified scoring system that avoids the potential problems of early treatment bias. A valid and simple method of risk-adjustment for neonatal intensive care is important to ensure accurate assessment of quality of care. Such assessments should be done in tandem with national audit systems for neonatal intensive care, incorporating measures of morbidity as well as mortality. PMID- 12781541 TI - Genital Chlamydia trachomatis infection in a subgroup of young men in the UK. AB - The frequency of genital Chlamydia trachomatis infection in young men in the UK has been found to be consistently lower than that in young women, but studies in such populations might have been affected by selection bias. We tested 798 male military recruits for chlamydia as part of their routine medical examination at Glencorse barracks in Scotland. 78 (9.8%) men were infected with chlamydia; rates of infection were similar in all age-groups. 69 (88%) chlamydia-positive men were asymptomatic. This rate is higher than those usually cited, showing the importance of opportunistic testing for chlamydia in men as well as in women. PMID- 12781542 TI - WHA approves historic tobacco accord. PMID- 12781545 TI - New hope for AIDS vaccine. PMID- 12781546 TI - Researchers call for more research into asthma in Latin America. PMID- 12781543 TI - Telerobotics brings surgical skills to remote communities. PMID- 12781547 TI - USA fights Europe's ban on genetically modified food. PMID- 12781549 TI - US Senate approves AIDS bill. PMID- 12781550 TI - Democrats see hope in health-care reform. PMID- 12781551 TI - Haemophilias A and B. AB - The haemophilias are inherited disorders in which one of the coagulation factors is deficient. Although deficiencies of factor VIII (haemophilia A) and factor IX (haemophilia B) are well recognised, von Willebrand's disease is much more common. Rare defects can occur in any of the coagulation factors. In the past, men with haemophilia were likely to die in their youth. With advances in diagnosis, and especially with development of safe and effective treatment, affected individuals can now look forward to a normal life expectancy. Complications of the disorder, particularly the development of antibodies that make treatment ineffective, and of treatment, such as transfusion-transmitted infections, have taken a severe toll on these patients. The future holds the realistic possibility of gene therapy. However, we must not forget that haemophilia is a worldwide disorder that requires significant economic resources not available for the majority. PMID- 12781552 TI - Continuity of neonatal care. PMID- 12781553 TI - Polycystic ovary syndrome and endometrial carcinoma. AB - CONTEXT: An association between polycystic ovary syndrome (PCOS)and endometrial carcinoma was first suggested in 1949, 14 years after the original description of the syndrome. Since then several studies have been published that seem to support this association. The prescription of hormonal treatment to reduce the risk of this complication is supported by the Guidelines for Good Clinical Practice of the Royal College of Obstetricians and Gynaecologists, UK, the Health Information website of the National Library of Medicine, USA, and in textbooks of gynaecological oncology. STARTING POINT: A recent practice bulletin from the American College of Obstetricians and Gynecologists on the clinical management of PCOS (Obstet Gynecol 2002; 100: 1389-402) says that there is still no consensus on the "optimal progestin, duration and frequency of treatment to prevent endometrial cancer in women with PCOS". Chronic anovulation, obesity, and hyperinsulinaemia are all associated with PCOS as well as with endometrial carcinoma. It has been assumed that PCOS predisposes to endometrial cancer. However, the evidence for such an association is inconclusive. Although PCOS is associated with risk factors for endometrial cancer, it does not necessarily follow that the incidence or mortality from endometrial cancer is increased. WHERE NEXT: Large-scale studies of morbidity and mortality in unselected populations of women with PCOS are needed. Women with PCOS are increasingly aware of the possible risks, and it will be necessary to identify which of them, if any, are at increased risk and how this risk can be effectively reduced. PMID- 12781554 TI - Increasing burden of treatment in the acute coronary syndromes: is it justified? PMID- 12781555 TI - Judging health systems: reflections on WHO's methods. PMID- 12781556 TI - Interferons in relapsing remitting multiple sclerosis. PMID- 12781557 TI - Interferons in relapsing remitting multiple sclerosis. PMID- 12781558 TI - Interferons in relapsing remitting multiple sclerosis. PMID- 12781559 TI - Interferons in relapsing remitting multiple sclerosis. PMID- 12781561 TI - Interferons in relapsing remitting multiple sclerosis. PMID- 12781563 TI - Interferons in relapsing remitting multiple sclerosis. PMID- 12781564 TI - Unrelated kidney donors. PMID- 12781566 TI - Potential for sibutramine-yohimbine interaction? PMID- 12781567 TI - The role of the Israel and World Medical Associations. PMID- 12781569 TI - The role of the Israel and World Medical Associations. PMID- 12781570 TI - The role of the Israel and World Medical Associations. PMID- 12781571 TI - When does exposure of children to tobacco smoke become child abuse? PMID- 12781572 TI - Genes for schizophrenia. PMID- 12781575 TI - Granulocyte-macrophage colony-stimulating factor for Crohn's disease. PMID- 12781574 TI - Genes for schizophrenia. PMID- 12781577 TI - Use of mobile phones in hospital. PMID- 12781578 TI - Gut flora in health and disease. PMID- 12781579 TI - What is best for the patient? PMID- 12781581 TI - SARS--a clue to its origins? PMID- 12781582 TI - Posthumous sperm retrieval. PMID- 12781585 TI - Screening negatives. PMID- 12781587 TI - Loss of nicotinic acetylcholine receptor subunits alpha4 and alpha7 in the cerebral cortex of Parkinson patients. AB - Cerebral cortical cholinergic deficits, represented by a decrease in choline acetyltransferase activity, severe losses of nicotinic binding sites as well as cell degeneration in the basal forebrain can be observed in neurodegenerative diseases such as Parkinson's disease and Alzheimer's disease. The potential role of nicotinic acetylcholine receptor subunits as pharmacological targets for the treatment of cognitive deficits raises the question as to what extent these subunits are affected in neurodegenerative diseases. We here report on a significant decrease of the alpha4 and the alpha7 nicotinic acetylcholine receptor subunit in cortices of Parkinson patients which turns out to be similar to recent findings in Alzheimer patients. PMID- 12781588 TI - Autosomal recessive parkinsonism linked to parkin gene in a Tunisian family. Clinical, genetic and pathological study. AB - OBJECTIVES: To report clinical, pathological and genetic findings in a Tunisian kindred with autosomal recessive juvenile parkinsonism (AR-JP) linked to parkin gene. BACKGROUND: AR-JP has been mapped to chromosome 6q and is caused by several mutations of the parkin gene (Park 2). Pathological features in AR-JP are characterized by neuronal loss in substantia nigra (SN) without Lewy bodies (LB). PATIENTS AND METHODS: Three affected siblings with juvenile Parkinson's disease were studied. Pathological examination of the brain was performed in one of them. Linkage studies and mutation analysis of the parkin gene were performed. RESULTS: Clinical picture was characterized by the association of rest tremor, bradykinesia and rigidity. Parkinsonian signs markedly improved with levodopa treatment in the three siblings. Dystonia was observed in one patient and diurnal fluctuations of parkinsonian signs in another one. Linkage analysis showed homozygous haplotypes in patients as compared to unaffected individuals and mutation analysis of the parkin gene revealed a homozygous two-base AG deletion in exon 2 (101-102). Pathological examination of the brain in one patient showed marked loss of pigmented neurons with extraneuronal free melanin in the lateral and medial parts of the SN associated to a slight spongiosis and astrocytic gliosis. In the locus coeruleus, there was also loss of pigmented neurons without gliosis. No LB or neurofibrillary tangles were found neither by traditional nor by histo-immunological stainings. CONCLUSION: This Tunisian kindred with AR-JP linked to a micro-deletion of the parkin gene shows clinical similarities with the previously reported Japanese and European families. Pathological features of this kindred are compared to what has been reported in AR-JP families linked to large exonic deletions of this gene. PMID- 12781589 TI - SPECT abnormalities with unilateral arm dystonia in a young mentally retarded apprentice cook: contralateral thalamo-cortical dysfunction. AB - We report a young, mentally retarded apprentice cook with a 2-month history of right upper extremity dystonia, for whom diazepam therapy was efficacious. We evaluated brain perfusion by single photon emission tomography (SPECT) before and after diazepam treatment. The abnormal hyperperfusion in the left thalamus and hypoperfusion in the left frontal cortex were normalized on the second SPECT under the successful diazepam treatment. These findings were indicative of functional changes in the left thalamus and left frontal cortex. PMID- 12781590 TI - The hung-up knee jerk in Huntington's Disease. AB - The 'hung-up' knee jerk (HUKJ) is a specific, although rarely appreciated, clinical sign of Huntington's Disease. A HUKJ was present in 11 of 31 consecutive patients with Huntington's Disease seen in our practice. The clinical characteristics of these patients are described and an EMG recording of the HUKJ is presented from one patient. The history of the description of the HUKJ and its neurophysiology are briefly reviewed. PMID- 12781591 TI - Adult-onset neuronopathic form of Gaucher's disease: a case report. AB - We report a patient with Gaucher's disease (GD) developing prominent neurological abnormalities in adult life confirming the existence of an adult neuronopathic form of GD. In this adult-onset form, an akinetic-rigid syndrome poorly responsive to dopatherapy, supranuclear gaze palsy, myoclonic jerks, seizures, cerebellar ataxia, cognitive and psychotic disturbances are frequent manifestations. The widely used clinical classification seems inadequate since it does not consider this rare form of GD. Until further understanding of the pathogenesis of the disease is achieved it is not possible to predict accurately which patients will or will not have late-onset nervous system involvement. PMID- 12781592 TI - Early and pre-symptomatic neuropsychological dysfunction in the PPND family with the N279K tau mutation. AB - The N279K mutation on the tau gene of chromosome 17 leads to an inherited condition that involves pallido-ponto-nigral degeneration (PPND). Patients with PPND develop dementia, but the pattern and onset of cognitive dysfunction has not yet been delineated. Four affected patients underwent neurocognitive evaluation within the first 2 years of PPND motor onset; one of whom underwent five serial neurocognitive evaluations, and another who was not diagnosed with PPND until the third annual evaluation. Impaired letter fluency was found in the early stages of PPND and was also shown to precede the onset of motor symptoms by 2 years. Trail Making A (visual scanning and motor speed) and Trail Making B (divided attention) were impaired within the first 2 years of the disease in all but one patient, but this individual showed clinically significant decline on these tasks by the third year of the disease. Learning, memory, and timed visuospatial sequencing skills were variably affected. Results reveal disproportionate frontal-executive dysfunction early in PPND disease course, a pattern similar to what has been reported in other FTDP-17 kindreds and in sporadic PSP. In addition, results suggest that letter fluency may be a sensitive predictor of incipient PPND. PMID- 12781593 TI - The increased utilisation of dopamine agonists and the introduction of COMT inhibitors have not reduced levodopa consumption--a nation-wide perspective in Sweden. AB - OBJECTIVE: To study the impact of the new pharmacological treatment strategies in Parkinson's disease by analysis of the sales of dopaminergic drugs in Sweden 1990 2001. METHODS: Invoice statistics and statistics on prescription sales of dopaminergic drugs 1990-2001 were obtained from the 906 public pharmacies and the 89 hospital pharmacies in Sweden. The Swedish Diagnosis and therapy survey was used to study the diagnosis the drugs were prescribed for. RESULTS: During the period the sales expressed in DDD/1000 inhabitants and day increased from 1.75 to 1.86 (6%) for levodopa and from 0.04 to 0.27 (575%) for dopamine agonists. The increase in sales of dopamine agonists occurred after 1997 and consisted entirely of the sales of the new agonists, cabergoline, pramipexole and ropinirole. In Swedish crowns the total sales of dopaminergic drugs increased with 126% during the period. The cost for the dopamine agonists was 8% of the total cost for dopaminergic drugs in 1990 and 23% in 2001. The prescription sales figures of levodopa and dopamine agonists for the different Swedish counties in 2001 varied between 1.52 and 2.44 and 0.13 and 0.74 DDD/1000 inhabitants and day, respectively. There was no correlation, whether positive or negative, between the sales of dopamine agonists, levodopa and COMT inhibitors or between the sales and the densities of neurologists. CONCLUSION: The consumption of levodopa in Sweden continues to increase in spite of a dramatic increase in the utilisation of dopamine agonists and the introduction of COMT-inhibitors. PMID- 12781594 TI - Unusual phenotypic expression of the DYT1 mutation. AB - Highly variable phenotype expression has long been recognized in DYT1 carrier patients. We report here an Ashkenazi-Jewish woman who carried a DYT1 mutation and developed a predominant unilateral myoclonic-dystonia (MD) displaying a fluctuating course. The present case is the second supporting the variability of DYT1 phenotype and further illustrates its ability to mimic the MD syndrome. PMID- 12781595 TI - Tics and developmental stuttering. AB - BACKGROUND: Developmental stuttering affects 1% of the population but its cause remains unclear. Recent PET studies of metabolism in the central nervous system suggest that it may be related to dysfunction in the basal ganglia or its connections with regions of the cortex associated with speech and motor control. OBJECTIVE: To determine the presence and characteristics of involuntary movements (IMs) in people who stutter and to investigate the hypothesis that these movements may be of a very similar nature to the IMs seen in patients with movement disorders due to basal ganglia dysfunction. METHODS: Sixteen adults with developmental stuttering and 16 controls matched for sex and age were audio videotaped while freely speaking 300 words in conversation and reading aloud 300 words. The audio data was inspected for dysfluencies and the video data was scrutinised for the presence and characteristics of IMs. RESULTS: Subjects who stuttered produced more IMs than controls during free speech (354 vs 187, p<0.05) and reading (297 vs 47, p<0.001). Most of the IMs in both groups were tics, with a greater number of both simple and complex motor tics (CMTs) in subjects who stuttered. CMTs were more frequent than simple motor tics in those who stuttered, but not in controls. The combination of repetitive eye blink followed by prolonged eye closure was found exclusively in the stuttering group, as were simple tics consisting of eyebrow raise or jaw movement. Dystonia in the form of blepharospasm was identified in a small number of subjects who stuttered. Choreic movements were not associated with stuttering. CONCLUSIONS: Developmental stuttering is associated with the presence of IMs that are predominantly simple and CMTs. This association suggests that tics and stuttering may share a common pathophysiology and supports the view that, in common with tics, stuttering may reflect dysfunction in the basal ganglia or its immediate connections. PMID- 12781596 TI - The psychometric properties of the Parkinson's Impact Scale (PIMS) as a measure of quality of life in Parkinson's disease. AB - We have previously designed and validated a 10-item bilingual questionnaire, the Parkinson's Impact Scale (PIMS), as a disease-specific instrument to measure the impact of Parkinson's disease (PD) on the quality of life of patients with PD. In this paper we extend the psychometric assessment of PIMS to a new set of patients, in the context of a cross-over trial by Hoffman-La Roche Ltd, comparing two doses of tolcapone in 116 PD patients who had developed wearing off effect on levodopa. Using data from this trial, we evaluate PIMS' test-retest reliability, construct validity, sensitivity and responsiveness to change. Validation is carried out by correlating the PIMS scores with corresponding UPDRS subscales and with the Schwab and England scale. We show that PIMS has excellent psychometric properties, and can therefore be used not only in clinical trials but also to identify quickly potential problems in major subjective areas of PD patients' lives, in order to refer them effectively to appropriate providers of assistance. PMID- 12781597 TI - Pulse width is associated with cognitive decline after thalamic stimulation for essential tremor. AB - The present study sought to identify predictors of cognitive decline after thalamic deep brain stimulation (DBS) for essential tremor (ET). Twenty-seven patients (55%) with ET demonstrated mild cognitive decrements relative to pre surgical baseline (ET-D), whereas 22 patients (45%) were classified as neuropsychologically stable (ET-S). The ET-D and ET-S groups were comparable in terms of baseline demographic, disease, and neuropsychological characteristics, as well as post-surgical motor outcomes. However, the ET-D group had significantly higher pulse width (PW) stimulator settings, and a greater proportion of ET-D than ET-S patients underwent left in comparison to right thalamic stimulation. A subsequent step-wise discriminant function analysis revealed that disease onset after age 37 years and higher PW settings (>or=120 micros) were the strongest predictors of post-surgical cognitive decline in this sample. Findings indicate that although relatively higher PW settings might afford optimal tremor control in some patients, the corresponding risk of mild, probably often subclinical, cognitive morbidity must be weighed accordingly. PMID- 12781598 TI - The economic evaluation of pharmacotherapies for Parkinson's disease. AB - As well as the significant clinical effects of Parkinson's disease (PD), the disease places a high economic burden on society. Given the scarcity of health care resources, it is becoming increasingly necessary to demonstrate that new therapies for PD provide value for money in comparison with other potential interventions. This paper outlines the basic techniques of cost-effectiveness analysis and its application to PD. These techniques are illustrated by a recent economic evaluation of entacapone for use in Canada. PMID- 12781599 TI - Parkin mutations are rare in patients with young-onset parkinsonism in a US population. AB - Parkin mutations have been associated with an autosomal recessive-juvenile parkinsonism, but it is unclear how common such patients are in a US based clinic population. In this study, we screened for parkin gene mutations in a clinic based series of 27 consecutive patients in the US with typical parkinsonism beginning before the age of 50 years. Two patients began the disease before the age of 30 years. Among the 27 patients screened, only one patient, whose disease began at the age of 24 years, was found to harbor a parkin mutation (a 40 bp deletion in exon 3). In addition, we also identified four new polymorphisms in the open reading frames of the parkin gene in this population. Our results suggest parkinsonism due to mutations in the parkin gene is extremely rare in the US population when the disease begins over the age of 30. PMID- 12781600 TI - Sensory neuropathy in autosomal recessive juvenile parkinsonism (PARK2). AB - Autosomal recessive juvenile parkinsonism (ARJP/PARK2) is a distinct clinical and genetic entity characterized by early-onset levodopa-responsive parkinsonism, foot dystonia, sleep benefit, and hyperactive tendon reflexes. We report a patient with genetically confirmed ARJP, who showed mild sensory disturbance and diminished deep tendon reflexes in the advanced stage. Nerve conduction studies revealed a sensory dominant neuropathy, which has not been described in ARJP. We suggest that peripheral neuropathy may occur in patients with advanced ARJP due to the loss of parkin protein function, although the function of parkin in the peripheral nervous system remains to be clarified. PMID- 12781604 TI - Scoring in burned patients. our opinion. PMID- 12781605 TI - What is the prevalence of hypertrophic scarring following burns? AB - Hypertrophic scarring after burns remains a major problem and is considered to be "common". Pressure garments are commonly used as treatment even though there is little sound data that they reduce the prevalence or magnitude of the scarring. In 1999 we began a study of the efficacy of pressure garments on forearm burns. After studying 30 patients, mainly white adults, we found no hypertrophic scar in either those treated with pressure or without. This prompted us to review the literature on the prevalence of hypertrophic scarring after burns and found only four articles with a relatively small number of patients and only three geographical locations. It became clear that the prevalence of hypertrophic scarring is really unknown. We then did a retrospective study of 110 burn survivors and counted all hypertrophic scars of all sizes and locations in all races and found the prevalence hypertrophic scarring to be 67% which conflicts with the published reports and our prospective study and suggests that further research is necessary. We concluded that a worldwide, prospective survey is necessary to establish the prevalence of hypertrophic scarring after burns. In this article we are calling for and offering to organize this survey. PMID- 12781606 TI - Implications for Burns Unit design following outbreak of multi-resistant Acinetobacter infection in ICU and Burns Unit. AB - We reviewed the emergence of 13 cases of multi-resistant Acinetobacter infection in burns patients over a 12-month period. The outbreak was started in a non-burn patient in the intensive care unit (ICU) that spread to burns patients in ICU and then the Burns Unit. The importance of opportunistic infection, potential risk factors, treatment and clinical outcome of Acinetobacter infection in burns patients from this cluster of cases is described. This paper implicates the movement of burns patients and medical equipment between ICU and the Burns Unit in the spread of this infection. Future design of Burn Units should aim to incorporate features to allow the management of all burns cases in one location with all intensive care, burns and theatre facilities built in close proximity. PMID- 12781607 TI - Effects of rolling inhibition on smoke inhalation injury. AB - Inhalation of chemical and particulate products of smoke is one of the principal determinant of mortality following burn injury. Inflammatory responses have been implicated in the pathogenesis of lung injury after smoke inhalation. In the current study, we tested the inhibitory effect of Fucoidin on the neutrophil rolling stage of inflammatory response and determined the degree of pulmonary injury. Fifteen rats were divided into three groups: sham group (N: 5) of rats inhaled room air; control group (N: 5) inhaled smoke, and experimental group inhaled smoke and received Fucoidin. All the rats were sacrificed 24h after smoke inhalation. The trachea and lungs were removed totally; samples for histopathological and biochemical (myeloperoxidase (MPO)) analysis were taken from each lung and trachea. Morphologic studies using light and electron microscopes showed a decrease in lung parenchymal and tracheoepithelial injury in the experiment group of rats. Also, biochemical analysis of tissue MPO was significantly lower in test group than in control group. These results suggest that the inhibition of neutrophil rolling leads to a reduction of neutrophil invasion to pulmonary parenchyma and trachea, which may be beneficial for attenuating neutrophil mediated inhalation injury. PMID- 12781608 TI - Soluble IL-4 receptor improves the skin-graft-associated cytomegalovirus infection in thermally injured mice. AB - The susceptibility of thermally injured mice (TI-mice) to murine cytomegalovirus (MCMV) infection is markedly influenced by burn-associated type 2 T cell responses, which are common with severe thermal injuries. In the present study, the effect of soluble IL-4 receptor (sIL-4R) on the skin-graft-associated MCMV infection was investigated. The marked growth of MCMV was demonstrated in the salivary glands of TI-mice grafted with MCMV seropositive [MCMV sero(+)] skin. However, the growth of MCMV was not demonstrated in the salivary glands of TI mice grafted with MCMV sero(+) skin and treated with 50ng per mouse of sIL-4R. Compared with grafted normal mice, production of type 1 cytokines was markedly decreased when splenic T cells from TI-mice grafted with MCMV sero(+) skin were stimulated with anti-CD3 monoclonal antibody (mAb). The impaired type 1 cytokine production was recovered in cultures of splenic T cells from grafted TI-mice previously treated with sIL-4R. After grafting with MCMV sero(+) skin, the growth of MCMV was markedly inhibited in the salivary glands of severe combined immunodeficient (SCID) mice inoculated with T cells from TI-mice treated with sIL 4R. These results suggest that sIL-4R regulates the skin-graft-associated MCMV infection in TI-mice. PMID- 12781609 TI - Corticosteroid-impairment of healing and gastric pentadecapeptide BPC-157 creams in burned mice. AB - The amelioration of corticosteroid-impairment of healing by a stable gastric pentadecapeptide BPC-157 (GEPPPGKPADDAGLV, M(w) 1419, currently in early clinical trials for inflammatory bowel disease) was studied in thermally injured mice. Its effects on corticosteroid impaired healing of deep partial skin thickness burns, and burn-gastric lesions were investigated. Male NMRI-Hannover mice (sacrificed at 1-3,7,14 and 21 days following burning 20% of total burn area at the back (open flame for 7s) received intraperitoneally (per kg bw) 6alpha methylprednisolone (Depo-medrol, 1.0 or 10.0mg), or an equal volume of saline (5.0 ml), once daily, first application 30 min after injury, last 24h before sacrifice. The injury was subsequently treated by topical application of a thin layer of pentadecapeptide BPC-157 cream at three different levels a neutral cream of no treatment. Pentadecapeptide BPC-157 consistently improved given burn healing (both microscopical and tensionmetry assessment), and counteracted corticosteroid-impairment of burn healing. In burn-gastric lesions investigation of the effects of BPC showed an anti-ulcer effect of its own in burned non corticosteroid-treated mice and potentiated the anti-ulcer effect observed in 6alpha-methylprednisolone-treated mice. Pentadecapeptide BPC-157 inhibited corticosteroid immunosuppression. In vitro, in spleenic cells assessment, animals (sacrificed at day 21) treated with 6alpha-methylprednisolone 1mg showed decreased reactivity to nitrogen in comparison with control, healthy animals, while the addition of BPC-157 (1 microg/g cream) returned cell reactivity to values noted in control healthy animals. PMID- 12781610 TI - Influence of local anaesthetics on inflammatory activity postburn. AB - Most studies investigating the pathophysiological processes taking place inside an experimental burn wound use in vitro techniques, which only allow for fragmented measurements of the actual and complex processes occurring inside a burn wound in vivo. In the present study, which used a recently developed in vivo technique in the rat, a full-thickness burn was induced and resulted in the formation of a subcutaneous gelatinous edema with distinct borders to the surrounding connective tissue and free communication with the systemic circulation allowing it to be easily separated for further analysis. In the present study, we investigated the effects of topical local anaesthetics (EMLA) on the inflammatory cascade of a burn wound in vivo. Results showed significantly higher myeloperoxidase (MPO) levels in EMLA-treated burned animals (P<0.01) versus placebo-treated burned controls. EMLA treatment induced a significant inhibition of the synthesis of leukotrien B(4) (LTB(4)) (P<0.001), prostaglandin E(1) (PGE(1)) (P<0.001), prostaglandin E(2) (PGE(2)) (P<0.001) and thromboxane B(2) (TXB(2)) (P<0.001) versus control, while free radical formation did not differ significantly between EMLA-treated and control animals. In conclusion, topical local anaesthetics significantly inhibit the release of several mediators known to take important part in the pathophysiological events ensuing a burn injury, such as activation of pain mechanisms (PGE), oedema formation (LTB), and postburn ischemia (TXB). The increased numbers of leukocytes (MPO) in the burn wound induced by topical local anaesthetic treatment could suggest increased influx and/or increased viability of leukocytes postburn. PMID- 12781611 TI - Delayed dilated cardiomyopathy for major burn injuries. AB - PURPOSE: The early impact on the heart of severe burns has all been well documented previously. Here, we report on the late effects of burns upon the heart, and dilated cardiomyopathy, such aspects having been little reported previously in the literature. METHODS: Over the past 8 years (June 1991 to December 1998), 55 scalded or flame-burnt patients for whom the total burnt body surface area (TBSA) exceeded 50% were treated. Seventeen patients survived and were followed up. The mean age was 44 years. The mean follow-up period was 47.3 months; four patients displayed a previous history of heart disease. We used several parameters to evaluate and follow cardiac status to see the late effect of burns upon the heart. RESULTS: Of 17 patients, 3 patients (17.7%) were found to exhibit dilated cardiomyopathies after a mean time of 6 months post-injury. A dramatic recovery from symptoms and roentgenographic findings were observed for all patients following symptomatic treatment. CONCLUSION: Here we report three cases of delayed dilated cardiomyopathy, a condition that has been little reported in the past. There may be many causating factors. Several recommendations are described. PMID- 12781612 TI - Flammable liquid burns in children. AB - OBJECTIVES: To document and describe the effects of flammable liquid burns in children. To identify the "at risk" population in order to tailor a burns prevention programme. DESIGN, PATIENTS AND SETTING: Retrospective study with information obtained from the departmental database of children treated at the burns centre at The Royal Children's Hospital, Brisbane between August 1997 and October 2002. MAIN OUTCOME MEASURES: Number and ages of children burned, risk factors contributing to the accident, injuries sustained, treatment required and long-term sequelae. RESULTS: Fifty-nine children sustained flammable liquid burns (median age 10.5 years), with a clear preponderance of males (95%). The median total body surface area burned was 8% (range 0.5-70%). Twenty-seven (46%) of the patients required debridement and grafting. Hypertrophic scars occurred in 56% of the children and contractures in 14%, of which all of the latter required surgical release. Petrol was the causative liquid in the majority (83%) of cases. CONCLUSIONS: The study identified the population most at risk of sustaining flammable liquid burns were young adolescent males. In the majority of cases these injuries were deemed preventable. PMID- 12781613 TI - Epidemiology of burns admitted to Ain Shams University Burns Unit, Cairo, Egypt. AB - This study was prospectively carried out on 880 acutely burned patients admitted to the Burn Unit of Ain Shams University in Cairo, Egypt, during the period from the 1 May 1995 to the 31 October 2001, with the objective to analyze the epidemiological features of burned patients in our country. The means of age and percentage total body surface area burned (TBSB) were 27.8+/-2.9 years and 32+/ 5.7%, respectively. Most of the patients were city dwellers and were self referred to the Burn Unit. There were no significant yearly variations in hospital admissions. Seasonal variations in the form of peaks in winter and spring were noticed. Females (53.1%) were more prevalent than males (46.8%). Adults (61%) superceded children (39%). Male children (42.7%) population exceeded female children (35.8%). In contrast, adult females (64.1%) surpassed adult males (57.2%). The commonest burn location was home followed by outdoors then, work locations. Females were most commonly burned at home and mainly suffered from flame and scald burns. Males were most commonly burned in outdoor and work locations than at home and mainly sustained electric and flame burns. There was no difference in the mean percentage TBSB and burn depth in both sexes. Children were mostly burned at home and were mainly victims of scald and flame burns. They had lower rates of full thickness burns compared to adults. Adults were mostly burned in outdoor and work locations and mainly sustained flame and electrical injuries. The results of this study clearly highlighted the specific epidemiological features of burned patients in our area, and thus should provide the necessary information to develop proper burn prevention programs, thereby reducing the frequency of burns and burn-related deaths. PMID- 12781614 TI - Analysis of wax burn cases presenting to an Emergency Department in Hong Kong during the Chinese Mid-Autumn festival. AB - Records of patients presenting to the Emergency Department (ED) of the Prince of Wales Hospital with wax burns during Mid-Autumn festival from 1998 to 2001 were retrieved and analyzed. There were 26 patients in total (21 males, 5 females). Annual attendances from 1998 to 2001 were 11, 7, 4 and 4, respectively. Age ranged from 1 to 33 years (median age: 11.5). Most patients were injured by molten wax (n=23, 88.5%), the rest were burnt by flame (n=3, 11.5%). Partial thickness burn (superficial and deep) was the most common (n=23, 88.4%). Superficial burn accounted for the rest (n=3, 11.5%). No patient had full thickness burn. All patients had <5% of body surface area (BSA) burnt, with the majority only involving <1% BSA (n=16, 61.5%). The commonest sites of injury were the lower limbs (n=15, 57.7%), the upper limbs (n=8, 30.8%) and face (n=7, 26.9%). Three patients (11.5%) had multiple sites burnt. Only eight patients (30.8%) required burns surgeon's consultation, out of which seven (26.9%) required in-patient treatment. Most burns caused by or related to boiling wax were minor. The declining incidence is the combined result of legislation, product modification, education and publicity. PMID- 12781615 TI - A retrospective study to determine the incidence of pressure ulcers in burn patients using a low air loss pressure relieving mattress. AB - In immobilized patients, unrelieved pressure can create decubitus ulcers over bony prominences. Those burn patients who require prolonged bed rest, are prone to the development of such problems. Various methods of reducing pressure on these areas, including frequent turning and the use of air fluidized and low air loss beds, have been adopted to attempt to prevent the development of this complication. The Pegasus Renaissance alternating pressure mattress is such a device, intended to reduce the incidence of decubitus ulcers. It was introduced at our burn unit and evaluated over a 29-month period. During the study period, 186 (13.4%) of 1390 acutely burned patients, believed to be at high risk for the development of decubiti, were placed on this mattress. Other patients were treated in the standard hospital bed. Care was otherwise the same. No decubitus ulcers developed in any of the patients treated on the Pegasus Renaissance mattress. PMID- 12781616 TI - Incidence of cardiac events in burned patients. AB - Given the increased level of adrenergic stimulation in burn patients, it would be expected that they would experience an increased incidence of cardiac arrhythmias and other cardiac events. We performed a retrospective chart review of 56 acute burn patients matched by age, length of hospital stay, and sex to 56 trauma patients, all of whom had been continuously monitored electrocardiographically. Burn and trauma patients were similar in injury severity, admission laboratory values, and prior history of cardiopulmonary diseases. Arrhythmias were noted in 34% of burn patients and 28% of trauma patients. One myocardial infarction and six deaths occurred in burn patients. No myocardial infarctions or deaths were observed in trauma patients. A past history of cardiopulmonary disease increased the risk of myocardial infarction or death by 6.6 times. Cardiac arrhythmias and other events are relatively infrequent and benign in burn patients and are similar to those experienced by other patients with acute injuries. PMID- 12781617 TI - "Amnion Bank"--the use of long term glycerol preserved amniotic membranes in the management of superficial and superficial partial thickness burns. AB - With about 2 million cases of burns in India of which about 90% are superficial and superficial partial thickness burns, burn injuries present a major challenge to the scarce medical resources available. The main requirement in treating these burns is an economical, easy to apply, readily available dressing that will provide good pain relief, protect the wound from infection, promote healing, prevent heat and fluid loss, be elastic and non-antigenic and adhere well to the wound. It was in the quest of such an ideal dressing that the Burn Centre of the Army Hospital (Research and Referral) New Delhi, India has been using human amniotic membranes preserved long term in 85% glycerol for treating these wounds. The results of this quest have resulted in a protocol where these long term glycerol preserved membranes (some of which have now been preserved for about 2.5 years) are used for treating superficial and superficial thickness burns presenting at our center. This paper shows the excellent results obtained with the use of this extremely economical dressing and emphasizes the importance of establishing such "Amnion Banks" in all hospitals especially in developing countries. PMID- 12781618 TI - Skin replacement with a collagen based dermal substitute, autologous keratinocytes and fibroblasts in burn trauma. AB - This case report describes the successful wound management of a 19-year-old patient with 76% of his body surface burnt. Due to the severity of the burns and the limited availability of native skin, 63% of the body surface were covered with Integra, a dermis equivalent. This material has been developed for the reconstruction of dermal structures and is designed as a bilayer. It consists of a modified, bovine collagen matrix and a silastic membrane facing toward the body surface. After vascularization of Integra, the epidermal skin replacement was created in a second step. In four different therapy procedures autologous, cultivated keratinocytes were combined with autologous fibroblast suspensions of various concentration and split-thickness skin as a 1:6 mesh graft. Rapid and largely complication-free wound healing could be achieved despite the limited availability of native skin. A correlation could be observed between the time of complete wound closure and the number of fibroblasts applied. The patient was monitored over a period of 1 year and showed good functional and cosmetic results. PMID- 12781619 TI - Proliferative "crescentic" glomerulonephritis in a burned patient. AB - Acute renal failure is one of the major complications of burn and it is accompanied by a high mortality rate. However, acute glomerulonephritis due to major burn have not been reported in burn literature. We report a case of crescentic glomerulonephritis which began at 27 days postburn. In this case glomerulonephritis may be due to infection probably pseudomonas or enterococus sepsis. We also felt that imipenem may be contributed the formation of glomerulonephritis. PMID- 12781620 TI - Combined 99mTc MDP bone SPECT and 99mTc sestamibi muscle SPECT for assessment of bone regrowth and free muscle flap viability in an electrical burn of scalp. AB - A case of deep high energy electrical burn of the skull is presented. For assessment of bone regrowth and muscle flap viability after application of latissimus dorsi free flap, combined 99mTc MDP bone SPECT and 99mTc sestamibi muscle SPECT scintigraphy were used. Whereas 99mTc MDP bone SPECT showed absent uptake at the beginning, there was good uptake 3 months after flap application. 99mTc sestamibi SPECT revealed good uptake indicating normal perfusion of flap muscle tissue. PMID- 12781621 TI - Survival of severe ARDS with five-organ system failure following burns and inhalation injury in a 15-year-old patient. AB - OBJECTIVE: To show the effectiveness of an integrated therapeutical approach in a severe case of acute respiratory distress syndrome (ARDS) following burns, inhalation injury with therapy-refractory oxygenation under maximized ventilatory settings, and an overall complicated clinical course. PATIENT AND METHODS: Case report of a patient with severe inhalation injury and burns in an intensive care unit setting, undergoing cardiopulmonary resuscitation (CPR), nitric oxide (NO) inhalation, surfactant-, kinetic-, and urodilatin-therapy. CASE REPORT: A 15-year old male presented with deep dermal and full thickness thermal injuries involving 25% of his total body surface area. Shortly after presentation, the patient developed therapy-refractory respiratory failure, cardiac arrest, and subsequently suffered five-organ system failure (lung, heart, gastrointestinal, liver, kidney), in addition to burn injury, and ischemia related cerebral lesions. The patient was successfully treated with cardiac resuscitation, extra corporeal membrane oxygenation (ECMO), NO, kinetic therapy, surfactant, urodilatin, and other standard intensive care regimens. Three months post-trauma the patient was discharged home, nearly fully recovered. CONCLUSIONS: In a patient with severe ARDS, oxygenation failure under maximized ventilatory settings, and subsequent five-organ system failure, an integrated therapeutical approach comprising ECMO, NO, kinetic therapy, surfactant, and urodilatin did cross-bridge respiratory and vital functions, enabling overall survival. PMID- 12781622 TI - Delayed obturator artery rupture: a complication of high-voltage electrical injury. AB - Electrical burns cause damage to the cardiovascular system through different mechanisms. Immediate cardiac arrhythmia is one of the common consequences. A large vessel blow out due to an electrical burn is rarely documented. It is often undetected due to its "silent" symptoms. This article reports a delayed obturator artery rupture in a 32-year-old male patient 6 weeks after he sustained severe high-voltage electrical burns. The vessel injury was treated with computed tomography (CT)-aided embolization. The patient recovered to an ambulatory status 10 weeks after the injury. PMID- 12781623 TI - A case of extensive second degree burn caused by ultraviolet ray irradiation. PMID- 12781624 TI - Potassium permanganate burn due to a dispensing error. PMID- 12781625 TI - Speciation of arsenic in human nail and hair from arsenic-affected area by HPLC inductively coupled argon plasma mass spectrometry. AB - Nail and hair are rich in fibrous proteins, i.e., alpha-keratins that contain abundant cysteine residues (up to 22% in nail and 10-14% in hair). Although they are metabolically dead materials in the epidermis, the roots are highly influenced by the health status of the living beings and their analyses are used as a tool to monitor occupational and environmental exposure to toxic elements. The aims of the present study are to speciate arsenicals in human nail and hair and also to judge whether they should be used as a biomarker to arsenic (As) exposure and/or toxicity. All human fingernail and hair samples (n = 47) were collected from the As-affected area of West Bengal, India. Speciation of arsenicals in water extracts of fingernails and hair at 90 degrees C was carried out by HPLC-inductively coupled argon plasma mass spectrometer (ICP MS). Fingernails contained iAs(III) (58.6%), iAs(V) (21.5), MMA(V) (7.7), DMA(III) (9.2), and DMA(V) (3.0), and hair contained iAs(III) (60.9%), iAs(V) (33.2), MMA(V) (2.2), and DMA(V) (3.6). Fingernails contained DMA(III), but hair did not. The higher percentage of iAs(III) both in fingernails and hair than that of iAs(V) suggests more affinity of iAs(III) to keratin. Although all arsenicals in fingernails and hair correlate to As exposure positively, As speciation in fingernails seems to be more correlated with arsenism than that in hair. Exogenous contamination is a confounding factor for hair to consider it as a biomarker, whereas this is mostly absent in fingernails, which recommends it to be a better biomarker to arsenic exposure. DMA(III) content in fingernails and DMA(V) contents in both fingernails and hair could be the biomarker to As exposure. PMID- 12781626 TI - Investigations on the inflammatory and genotoxic lung effects of two types of titanium dioxide: untreated and surface treated. AB - TiO(2) is considered to be toxicologically inert, at least under nonoverload conditions. To study if there are differences in lung effects of surface treated or untreated TiO(2) we investigated the inflammatory and genotoxic lung effects of two types of commercially available TiO(2) at low doses relevant to the working environment. Rats were exposed by instillation to a single dose of 0.15, 0.3, 0.6, and 1.2 mg of TiO(2) P25 (untreated, hydrophilic surface) or TiO(2) T805 (silanized, hydrophobic surface) particles, suspended in 0.2 ml of physiological saline supplemented with 0.25% lecithin. As control, animals were instilled with the vehicle medium only or with a single dose of 0.6 mg quartz DQ12. At days 3, 21, and 90 after instillation bronchoalveolar lavage was performed and inflammatory signs such as cells, protein, tumor necrosis factor alpha, fibronectin, and surfactant phospholipids were determined. Additionally, 8 microm frozen sections of the left lobe of the lung were cut and stored at -80 degrees C. The sections were used for immunohistochemical detection of 8 oxoguanine (8-oxoGua) by a polyclonal antibody in the DNA of individual lung cells. In the quartz-exposed animals a strong progression in the lung inflammatory response was observed. Ninety days after exposure a significant increase in the amount of 8-oxoGua in DNA of lung cells was detected. In contrast, animals exposed to TiO(2) P25 or TiO(2) T805 showed no signs of inflammation. The amount of 8-oxoGua as a marker of DNA damage was at the level of control. The results indicate that both types of TiO(2) are inert at applicated doses. PMID- 12781627 TI - Effects of endocrine disruptors on arachidonic acid metabolism in rabbit platelets. AB - To explore the possible actions of endocrine disruptors on the autacoid synthesis in the body, we investigated the effects of nonylphenol (NP), bisphenol A (BPA), di-n-butyl phthalate (DBP), benzyl-n-butyl phthalate (BBP), and di-2-ethylhexyl phthalate (DEHP) on the formation of 12-lipoxygenase metabolite, 12-HETE, and cyclooxygenase metabolites, TXB(2) and 12-HHT, from exogenous arachidonic acid (AA) in rabbit platelets. NP (10-50 microM) showed strong inhibition on the formation of cyclooxygenase metabolites (TXB(2), 34-95% inhibition; 12-HHT, 13 78% inhibition) and weaker inhibition on the formation of 12-HETE (0-49% inhibition). BPA, DBP, BBP, DEHP, and 17beta-estradiol (endogenous estrogen) failed to show any effect on the formation of cyclooxygenase and 12-lipoxygenase metabolites at concentrations up to 100 microM. These results suggest that NP inhibits AA metabolism in platelets and that its effects on the cyclooxygenase pathway predominate over those exerted via the 12-lipoxygenase pathway. PMID- 12781628 TI - Characterisation of some cytotoxic endpoints using rat liver and HepG2 spheroids as in vitro models and their application in hepatotoxicity studies. I. Glucose metabolism and enzyme release as cytotoxic markers. AB - Cytotoxicity endpoints, spontaneous glucose secretion/consumption and LDH and gamma-GT release, were characterised in rat liver and HepG2 spheroids as in vitro models for toxicology studies. Preprepared rat liver spheroids and HepG2 spheroids cultured in a six-well plate format were exposed to varying concentrations of galactosamine, propranolol, diclofenac, and paracetamol. All four model toxins significantly affected glucose secretion, which agreed well with LDH and/or gamma-GT release in rat liver spheroids. These toxins also significantly increased LDH and/or gamma-GT release in HepG2 spheroids. Whereas glucose consumption in HepG2 spheroids did not show conclusive results, LDH activities in both types of spheroids were similar and their levels were relatively high. Accordingly, the level of LDH leakage in both types of spheroids was much higher than gamma-GT after exposure to the toxins. In contrast, gamma-GT activity in HepG2 spheroids was sixfold higher than that in rat liver spheroids. This study revealed that galactosamine interfered with the gamma-GT assay and paracetamol interfered with the LDH assay. It demonstrated, for the first time, that glucose secretion by liver spheroids can be used as a functional indicator of cytotoxicity. Test compounds may interfere with enzymatic assays as indicated by LDH and gamma-GT release in this study. Combining functional parameters together with two or more indicators of enzyme releases can provide a reliable cytotoxicity evaluation. Liver and HepG2 spheroids as in vitro models showed good predictions in chemical-induced hepatic cytotoxicity. PMID- 12781629 TI - Characterisation of some cytotoxic endpoints using rat liver and HepG2 spheroids as in vitro models and their application in hepatotoxicity studies. II. Spheroid cell spreading inhibition as a new cytotoxic marker. AB - Cells in liver spheroids and Hep G2 spheroids transferred from gyrotatory culture conditions and maintained in normal static culture conditions will spread out at the edges. Based on this observation, we developed a new test called the Spheroid Cell Spreading Inhibition Test (SCSIT) to screen hepatic cytotoxicity of xenobiotics and determine the spheroid cell spreading inhibition concentration (SCSIC) of test chemicals. Four model hepatoxicants, D-galactosamine, propranolol, diclofenac, and paracetamol, were studied with SCSIT in both rat liver and HepG2 spheroids. Both liver and HepG2 spheroids were prepared under gyrotatory culture conditions and used at 6 days in vitro. The results showed that all four hepatotoxicants tested inhibited cell spreading in liver spheroids (D-galactosamine at 20 mM, propranolol at 125 microM, diclofenac at 500 microM, and paracetamol at 25 mM) and HepG2 spheroids (D-galactosamine at 16 mM, propranolol at 125 microM, diclofenac at 500 microM, and paracetamol at 25 mM). The SCSIT results agreed with the conventional cytotoxic indicators, release of LDH and/or gamma-GT and the inhibition of glucose secretion from rat liver spheroids. In conclusion, this study, for the first time, described the biological characteristics of liver and HepG2 spheroid cell spreading and demonstrates its application in hepatic cytotoxicity studies. This method may be used in testing in vitro "acute" toxicity, comparing relative cytotoxicity and generating reference concentrations for subsequent studies. Therefore, SCSIT could be a useful tool for screening hepatotoxicity relevant to preclinical lead optimization and compound library screening. PMID- 12781630 TI - Oxazolone and diclofenac-induced popliteal lymph node assay reactions are attenuated in mice orally pretreated with the respective compound: potential role for the induction of regulatory mechanisms following enteric administration. AB - The murine popliteal lymph node assay (PLNA) was examined as a preclinical assay with the potential to identify low-molecular-weight compounds (LMWCs) that are likely to be associated with immune-mediated drug hypersensitivity reactions (IDHRs) in humans. We hypothesized that the contact sensitizer oxazolone (OX) would cause a strong PLN reaction in naive mice and that the PLN reaction would be attenuated in mice orally pretreated with OX due to the induction of oral tolerance. In naive mice, OX induced a strong PLN reaction and caused dose dependent increases in PLN size, weight, cellularity, percentage of CD4(+) PLN T cells, and percentage of PLN B cells, with a concomitant decrease in the percentage of CD8(+) PLN T cells. Next, the PLNA was conducted in mice gavaged three times with either OX or vehicle alone (olive oil). Mice pretreated with OX had suppressed PLN reactions following the footpad injection of OX (decrease in PLN size, weight, and cellularity), which was associated with an increase in the percentage of PLN CD8(+)T cells. In contrast, oral pretreatment with OX had no observable effect on the PLN reaction induced following footpad injection of the irrelevant hapten dinitrochlorobenzene (DNCB). Adoptive transfer studies were conducted to examine the mechanism of PLN hyporesponsiveness. It was found that either (1) unfractionated splenocytes or (2) purified CD8(+) splenocytes, but not (3) purified CD4(+) splenocytes isolated from mice gavaged with OX adoptively transferred PLN suppression to naive BALB/c mice. Because OX is not a pharmaceutical, we also examined the NSAID diclofenac (DF) (Voltaren). Like OX, DF caused dose-dependent increases in PLN size, weight, and cellularity in naive mice. Furthermore, like OX, the diclofenac-induced PLN reaction was attenuated in mice that had been orally pretreated three times with DF. However, splenocytes from mice orally treated with DF were not able to adoptively transfer PLN hyporesponsiveness. Collectively, these observations demonstrate that both OX and DF are potent immunostimulators in the PLNA. As importantly, these results demonstrate that the immunostimulating potential of OX and DF in the PLNA is significantly decreased in mice orally exposed to the respective drug, possibly due to the presence of a cellular mechanism of oral tolerance. For OX, the mechanism appears to involve, in part, CD8(+) T cells, whereas the mechanism(s) associated with PLN hyporesponsiveness using DF remain to be defined. PMID- 12781631 TI - Visually evoked cortical potentials obtained using checker patterns can detect ethambutol-induced visual toxicity in albino rats. AB - We determined whether visually evoked cortical potentials obtained using checker patterns (P-VECPs) and albino rats would reveal visual damage induced by ethambutol (EB). Findings were compared in cases of detection of visual damage between by P-VECPs and by flash visually evoked cortical potentials (F-VECPs). Twelve adult albino male Crj:CD(SD)IGS rats were grouped into four, three per group: control, 250PS, 500PS, and 500SC groups. In the 250PS and 500PS groups, rats were administered EB orally for the first 2 weeks and then subcutaneously for the second 2 weeks to 250 and 500 mg/kg, respectively. In the 500SC group, rats were given 500 mg/kg of EB subcutaneously for 4 weeks. Rats in the control group were given the vehicle orally for the first 2 weeks and then subcutaneouly for the second 2 weeks. P-VECPs and F-VECPs were carried out prior to initiation of drug administration and at the 1st, 2nd, 3rd, and 4th weeks of the administration. Prolongation of P1 latency in the P-VECPs was evident in both the 500PS and the 500SC groups at the 4th week, while no marked changes were observed in the F-VECPs. Thus, P-VECPs in albino rats can detect visual damage induced by EB even when F-VECPs cannot do so. These studies suggest that P-VECPs are useful for evaluating the visual toxicity of drugs. PMID- 12781632 TI - Parenteral N,N-diethyldithiocarbamate produces segmental demyelination in the rat that is not dependent on cysteine carbamylation. AB - Disulfiram, a dithiocarbamate drug used in alcohol aversion therapy, produces a peripheral neuropathy characterized in rats as segmental demyelination accompanied by generation of S-(diethylaminocarbonyl)cysteine (DETC-Cys) adducts. N,N-Diethyldithiocarbamate (DEDC) is a major metabolite of disulfiram that can undergo methylation and oxidation to S-methyl-N,N-diethylthiocarbamate (MeDETC) sulfoxide and sulfone, thought to be responsible for carbamylation of sulfhydryl functions by disulfiram. To assess the role of cysteine carbamylation in disulfiram toxicity, DEDC and MeDETC were administered parenterally to male Sprague-Dawley rats for 4 and 8 weeks. The roles of the disulfide linkage in disulfiram and of carbamylated glutathione metabolites were assessed by administering S-(diethylaminodithiocarbonyl)N-acetylcysteine (DS-NAC) and S (diethylaminocarbonyl)-N-acetylcysteine (DETC-NAC), respectively, parenterally for 12 weeks. Following exposure, spinal cord-derived neurofilament preparations and hemoglobin were isolated and analyzed by RP-HPLC and LC/MS/MS for the presence of DETC-Cys adducts. Peripheral nerve sections were also obtained and examined by light and electron microscopy for morphological lesions. RP-HPLC analysis of globin preparations from DEDC-, MeDETC-, and DS-NAC-exposed animals demonstrated a late-eluting peak, identical to that reported for disulfiram generated DETC-Cys adducts on the beta(3)-globin chain. DETC-NAC exposure did not result in detectable globin modification by RP-HPLC. The quantity of DETC-Cys adducts produced on globin and neurofilament preparations determined by LC/MS/MS was twofold greater for MeDETC than DEDC following equimolar doses of each compound. Primary myelin lesions consisting of demyelinated axons and myelin splitting were observed in peripheral nerves following exposure to DEDC for 8 weeks. No lesions were detected following exposure to MeDETC, DS-NAC, or DETC-NAC at any time point or dose level. These results are consistent with DEDC, but not the other metabolites, being a demyelinating agent and thus a potential proximate toxic species for disulfiram-mediated demyelination. The production of significantly greater levels of DETC-Cys adducts by MeDETC relative to DEDC in the absence of neurotoxicity for MeDETC is consistent with cysteine carbamylation not contributing to the demyelination produced by disulfiram and DEDC. PMID- 12781633 TI - A wholly nutritional 'multifocal angiostatic therapy' for control of disseminated cancer. AB - A great deal of effort is now being devoted to the development of new drugs that hopefully will control the spread of inoperable cancer by safely inhibiting tumor evoked angiogenesis. However, there is growing evidence that certain practical nutritional measures have the potential to slow tumor angiogenesis, and it is reasonable to anticipate that, by combining several measures that work in distinct but complementary ways to impede the angiogenic process, a clinically useful 'multifocal angiostatic therapy' (MAT) might be devised. Several measures which might reasonably be included in such a protocol are discussed below, and include: a low-fat, low-glycemic index vegan diet, which may down-regulate the systemic IGF-I activity that supports angiogenesis; supplemental omega-3-rich fish oil, which has been shown to inhibit endothelial expression of Flk-1, a functionally crucial receptor for VEGF, and also can suppress tumor production of pro-angiogenic eicosanoids; high-dose selenium, which has recently been shown to inhibit tumor production of VEGF; green tea polyphenols, which can suppress endothelial responsiveness to both VEGF and fibroblast growth factor; and high dose glycine, whose recently reported angiostatic activity may reflect inhibition of endothelial cell mitosis, possibly mediated by activation of glycine-gated chloride channels. In light of evidence that tumor-evoked angiogenesis has a high requirement for copper, copper depletion may have exceptional potential as an angiostatic measure, and is most efficiently achieved with the copper-chelating drug tetrathiomolybdate. If logistical difficulties make it difficult to acquire this experimental drug, high-dose zinc supplementation can achieve a slower depletion of the body's copper pool, and in any case can be used as maintenance therapy to maintain an adequate level of copper depletion. A provisional protocol is offered for a nutritionally based MAT entailing a vegan diet and supplemental intakes of fish oil, selenium, green tea polyphenols, glycine, and zinc. Inasmuch as cox-2 is overexpressed in many cancers, and cAMP can boost tumor production of various angiogenic factors as well as autogenous growth factors, adjunctive use of cox-2-specific NSAIDS may be warranted in some cases. PMID- 12781634 TI - New biological approach and solution to SIDS. AB - The toothlessness and, specifically, the oral gap which is present between the lower and upper gums in the oral cavity of the toothless baby, while only temporary anatomical deficiencies, are, nevertheless, very important ones. It is suggested herein that this phenomenon of toothlessness and temporary physical abnormality has the ability to cause sudden infant death syndrome (SIDS) in a significant number of babies in their early months. What follows is a presentation of a new biological approach to the underlying causes of SIDS and, in the Conclusion, concrete and practical suggestions are given for a completely new method to counteract and neutralize the threatening dangers of SIDS. PMID- 12781635 TI - Dopamine receptor hetero-oligomerization: a hypothesis for behavioral sensitization to psychostimulants. AB - Repeated psychostimulant pretreatment can induce behavioral sensitization. Many previous studies have demonstrated that receptors of the dopamine and other neurotransmitter systems are involved in the sensitization process, however, the precise mechanism of this interaction is still unknown. We propose the hypothesis that oligomerization of the dopamine receptors and analogs from other neurotransmitter systems may contribute, at least in part, to psychostimulant induced behavioral sensitization. Psychostimulant exposure can enhance central dopamine release, which can increase oligomerization of the dopamine receptors with their analogs. Receptor oligomerization can enhance the functional properties of the receptors and affect receptor degradation, which may be related to the behavioral augmentation which characterizes the sensitized state and produces the persistence of behavioral sensitization. The proposed receptor hetero-oligomerization model may provide a new direction in the exploration of the functionally critical sensitization phenomenon. PMID- 12781636 TI - The relationship of long term global temperature change and human fertility. AB - According to the United Nations, global fertility has declined in the last century as reflected by a decline in birth rates. The earth's surface air temperature has increased considerably and is referred to as global warming. Since changes in temperature are well known to influence fertility we sought to determine if a statistical relationship exists between long-term changes in global air temperatures and birth rates. The most complete and reliable birth rate data in the 20th century was available in 19 industrialized countries. Using bivariate and multiple regression analysis, we compared yearly birth rates from these countries to global air temperatures from 1900 to 1994.A common pattern of change in birth rates was noted for the 19 industrialized countries studied. In general, birth rates declined markedly throughout the century except during the baby boom period of approximately 1940 to 1964. An inverse relationship was found between changes in global temperatures and birth rates in all 19 countries. Controlling for the linear yearly decline in birth rates over time, this relationship remained statistically significant for all the 19 countries in aggregate and in seven countries individually (p<0.05). CONCLUSIONS: The results of our analyses are consistent with the underlying premise that temperature change affects fertility and suggests that human fertility may have been influenced by change in environmental temperatures. PMID- 12781637 TI - How does skin adapt to repetitive mechanical stress to become load tolerant? AB - Skin breakdown from mechanical stress application is a difficult health care problem for lower-limb amputees using prosthetic limbs. Post-operative treatments to encourage skin adaptation do exist, but are largely unsuccessful. Potentially, by understanding skin adaptation on a molecular level, appropriate biomolecules can be identified and then delivered to skin to encourage adaptation in at-risk patients. Based from a critical review of the literature, it is expected that adaptation occurs by forming new collagen fibrils with larger diameters as opposed to increasing diameters of existing fibrils. Small collagen fibril breakdown by stress activated metalloproteinases is expected to be followed by increased expressions of decorin, biglycan, fibromodulin, lumican, thrombospondin 2, and collagens I and III, facilitating formation of new fibrils with larger diameters. After remodeling, total collagen fibril cross-sectional area is expected to return to baseline values since increased collagen content would increase mass and be redundant towards the purpose of adaptation. PMID- 12781638 TI - Serotonin transporter gene functional polymorphism: a plausible candidate gene for increased vascular risk in depression. AB - The evidence of increased vascular morbidity and mortality associated with depression has generated research interest in studying the mechanisms or causal pathways underlying this association. Recent advances in molecular genetics have demonstrated that serotonin transporter gene functional polymorphism may confer susceptibility for affective disorder as well as for some cardiovascular risk factors. Taking into account these genetic findings, this article proposes a hypothesis that serotonin transporter gene functional polymorphism may be a plausible candidate gene to study the genetic mechanisms of depression-related increased vascular morbidity and mortality. Future research projects to test this hypothesis is warranted. PMID- 12781639 TI - Growth and cellular differentiation: a physico-biochemical conundrum? The example of the hand. AB - Currently, the predominant hypothesis explains cellular differentiation as an essentially genetic intracellular process. The goal of this paper is to suggest that cell growth and differentiation may be, simply, the result of physical and chemical constraints. Bone growth occurs at the level of cartilage conjunction (growth plate) in a zone of lesser constrain. It appears that this growth also induces muscle, tendon, nerve and skin elongation. This cartilage growth by itself seems to explain the elongation of the hand. Growth stops at puberty likely because of feed-back from an increasing muscle load. The ossification (that is differentiation of cartilage into bone) appears to result from the shear stress induced. The study of bone age, obtained by X-ray picture of the hand, shows that ossification of epiphyses is very precise both in time and space. Computer modelization suggests that this ossification occurs where shear stress is greatest. The cartilage which does not ossify (joint, nose, larynx, ear, bronchus, etc.) is not exposed to high shear. Shear stress induces the secretion of extracellular matrix and a change of the biochemical environment of the cell. Precipitation of calcium phosphate, as in ossification, seems related to the alkalosis induced by shear stress. To speak in more general terms, loss of cellular differentiation, as occurs with cancer, can result from a change in the physical-chemical environments. PMID- 12781640 TI - Atypical depression as a secondary symptom in chronic fatigue syndrome. AB - Chronic fatigue syndrome (CFS) has gained prominence since 1988 and a substantial amount of research has been done in this domain. However, it is still regarded as a controversial condition. Moreover, most of the symptoms of CFS itself are non specific, occurring in many illnesses; some of the symptoms are also common in depression. Indeed, an area of continued controversy and debate involves the diagnostic overlap between CFS and psychiatric disorders. Through anecdotal evidence, atypical depression appears to be common in CFS. Recent developments in psychobiology underscore the role of the acute phase response and its associated sickness behavior in affective disorders. Thus, we hypothesize that atypical depression is sickness behavior rather than an affective disorder as shown by anecdotal evidence in CFS. PMID- 12781641 TI - Biomathematical modeling for diluted drugs. AB - Several workers have proven that succussed ultra high dilution of a drug molecule in water or alcoholic medium, even exceeding Avogadro number, can bring forth noticeable physiological changes of an organism. Homeopathic drugs are prepared by dissolving such drug ingredients in distilled water and then the solution is centesimally diluted serially by ethanol. A mathematical model has been proposed by the present worker, which explains why the drug does not become non-molecular even in ultra-high dilution. This is due to loss of homogeneity in the solution, caused by increase of dielectric constant of the medium during the process of potentization. Facilitated binding of the drug molecules with minute physiologically important protein factors may be the cause of visible physiological alterations. PMID- 12781642 TI - The common mitochondrial DNA deletion deltamtDNA(4977): shedding new light to the concept of a tumor suppressor mutation. AB - We propose that the age-related accumulation of deltamtDNA(4977) mutations may serve a protective function against tumor-promoting effects of other somatic mutations. The evidence discussed here is consistent with the concept that deltamtDNA(4977) plays a tumor-suppressor role, thus shedding new light to the concept of a tumor suppressor mutation. This concept may help understand how a tumor-promoting mutation may be able to cause malignant transformation in cells lacking a tumor-suppressor mutation, while the same tumor-promoting mutation can be present in cells that carry a tumor-suppressor mutation, without causing cancer. PMID- 12781643 TI - Aberrant local renin-angiotensin II responses in the pathogenesis of primary sclerosing cholangitis. AB - Primary sclerosing cholangitis (PSC) is an idiopathic inflammatory disorder of the biliary tract characterized by diffuse biliary tract stricture formation, progressive chronic cholestasis and the development of secondary biliary cirrhosis. Biliary tract ischemia can produce morphological changes identical to PSC. We propose the existence of a localized renin-angiotensin system within the liver and extend the hypothesis that aberrant production of angiotensin II within the portal tract is the critical event contributing to the pathogenesis of PSC. A chronic reparative and proliferative state caused by chronic ischemia may promote carcinogenesis. Proof of this hypothesis will have implications for future therapeutic approaches given that current treatments for PSC aimed at reducing inflammation or the effects of cholestasis have proven ineffective. PMID- 12781645 TI - The novel mechanism of peripheral tolerance. AB - The circulating pool of lymphocytes contains self- as well as non-self reactive T cells. The mature DCs present as non-self as well as self-epitopes to nai;ve T cells. To avoid autoimmunity the organism has to keep the mature DCs afar from self-specific T cells. I had proposed that the different anatomical distribution of the immature (tolerogenic) and mature (immunogenic) DCs in the peripheral lymphoid tissues may contribute to this mechanism. I had proposed that T cells to reach the mature DC should pass through the layer of immature DCs, which constitutively phagocytose and transport apoptotic cells to the regional LN and present only 'self' epitopes. Thus, self-specific T cells will be trapped by the immature DC layer and be deleted or become anergic. The layer of immature DC will be crossed only by those T cells that are not self-specific. PMID- 12781644 TI - A novel terminology and dissemination theory for a subgroup of intrathoracic extrapulmonary hydatid cysts. AB - Chest wall, mediastinal, pericardial, myocardial, lobar fissure and pleural locations of the hydatid cysts are referred to be intrathoracic extrapulmonary cysts. In this report a 20-year-old male patient had a liver dome cyst, which was opened into the diaphragm and caused diaphragmatic hydatid cysts. The scoleces have probably been transported by diaphragmatic lymphatics into the subpleural location and then hydatid disease has been disseminated via parasternal lymph nodes anteriorly and intercostal lymph nodes posteriorly towards cupola of the thorax. With regard to this localization, we propose a novel terminology as 'intrathoracic extrapleural hydatid cyst' and we hypothesize a novel dissemination theory for the hydatid cysts via diaphragmatic lymphatic drainage. PMID- 12781646 TI - Associations between psychological profiles and diseases: examining hemispheric dominance and autonomic activation as underlying regulators. AB - Personality profiles are often typical for specific illnesses, e.g., the type A personality and heart disease. We hypothesize that many more such patterns exist, and have developed a scheme in which many diseases can be classified, depending on hemispheric dominance (i.e., integrated, intellectualising or emotional) and type of autonomic control (i.e., dominance of either sympathetic or parasympathetic system, or an increase in both types of autonomic outflow). Our hypothesis is based on recent findings in neurophysiology, involving the early rearing environmental effects on the developing orbitofrontal cortex, and attachment theory. We conclude with implications for therapy, and a discussion of the limitations of our hypothesis. PMID- 12781648 TI - Speculations about alternative DNA structures. AB - An alternative model to the Watson & Crick (W&C) double DNA-spiral and the Pauling & Corey (P&C) triple spiral is presented. In this model: (1). the rotation axis of the polynucleotide chain is in the ribose ring; (2). there is a H- bond or direct covalent bond between the O2 (PO(4) and C2(') (in ribose) which makes the nucleic acid strands 'stiff'; (3). when there is a covalent bond between O2 and C2('), the unit of the DNA is the ribonucleoside 2('), 3(')-cyclic monophosphate, an intermediate form between DNA and RNA; (4). the bases point outwards from the rotation axis and may interact with each other to connect 2-4 strands together through complementary base pairs; (5). two strands may, but do not necessarily, form a helical structure and if they do, the interacting strands do not turn around each other. The architecture of this model, termed the Homulus DNA model is open (in contrast to the inverted W&C model) and using it might help us to understand the nature of some specific DNA-protein interactions, ordered chromatin formation (coiling and de-coiling), specific gene-to-gene interaction (gene targeting). It is possible that a small portion of the total DNA, the transcribed, 'working DNA', might be built by this way. PMID- 12781647 TI - I/D ACE gene polymorphism in survival of leukemia patients -- hypothesis and pilot study. AB - Angiotensin-I converting enzyme (ACE) is involved not only in intracellular volume regulation but also in proliferation control. Since both ACE gene polymorphism (I/D ACE) and ABO blood group determine ACE level in peripheral blood and probably also in bone marrow, the hypothesis to the interindividual differences in survival of leukemic patients was suggested. The data of 25 patients of both sexes with acute myelogenous (AML), acute lymphatic (ALL), chronic myelogenous (CML) and chronic lymphatic (CLL) leukemia treated by conventional were used for the study. The overall survival (SUR) was estimated as the time from the date of diagnosis to the date of death. The difference between patient's individual SUR (iSUR) and median SUR according to the type of leukemia (mSUR) was calculated. This difference (iSUR-mSUR) varied with I/D ACE genotype (p<0.02) but neither with diagnosis nor with ABO blood group. The regression model for iSUR calculation, from mSUR and I/D ACE genotype, has been suggested. PMID- 12781650 TI - Toward a holistic-organismic conceptualization of attachment style. AB - A holistic-organismic approach is suggested as foundation for intensified, systematic inquiry into relationships among adult attachment styles and various dimensions of physical and mental health. Exploratory findings of differences in stamina associated with adult attachment style are noted. The extent to which attachment style-stamina associations may be related to other wellness-illness differences is considered. Illustrative components and directions in an ongoing program of research are outlined. PMID- 12781649 TI - Statins may potentiate bisphosphonates anticancer properties: a new pharmacological approach? AB - Both statins and bisphosphonates may inhibit cancer proliferation by two main different mechanisms: inducing apoptosis along cholesterol synthesis pathway and by antiangiogenic properties. Moreover, also an immunomediated mechanism could be involved in anticancer properties of these molecules. The association of these two drugs could represent an interesting pharmacological approach in anticancer treatment. The available data offer the rationale for future in vitro studies aimed at evaluating proapoptotic and antiangiogenic action of this association. If the results of vitro studies should confirm the hypothesis that statins potentiate the action of bisphosphonates, further clinical investigations could be mandatory to evaluate the efficacy of this new pharmacological approach in anticancer therapy. PMID- 12781651 TI - Brain edema -- a new classification. AB - Brain edema is a reaction to any brain injury and can be the first stage in the beginning of intracranial hypertension. This paper puts forth a modern classification of brain edema types, based on a etiopathogenic interpretation. The hydroelectrolitic and/or proteinic buildup can occur within cells and/or in the extracellular space and differentiates three types of brain edema: cellular brain edema; extracellular brain edema and combined brain edema. Cellular brain edema (cytotoxic brain edema) occurs through intracellular hyperosmolarity or extracellular hypotonicity. Extracellular brain edema (interstitial) appears as a result of the buildup of edema fluid in the extracellular space of the brain parenchyma and can be: hydrostatic extracellular brain edema (through ultrafiltration), oncotical extracellular brain edema (vasogen brain edema) and hydrocephalic extracellular brain edema. Combined brain edema includes in variable ratios both types of brain edema, cellular and extracellular; they can be present together from the beginning or can appear successively. PMID- 12781652 TI - Brain-derived neurotrophic factor: a bridge between major depression and Alzheimer's disease? AB - Cognitive impairment is common in major depression (MD) patients, with these individuals incurring increased risk for development of Alzheimer's disease (AD). Further, depressive symptoms are common in AD patients. This apparent convergence suggests pathogenic factors common to AD and MD. Since decreased brain-derived neurotrophic factor (BDNF), a member of the neurotrophic factor family, is related to both AD and MD, the author suggests that BDNF could be a bridge between AD and MD, explaining both the depressive symptoms in AD, and, cognitive impairment in MD. Evidence supporting this hypothesis suggests that early antidepressants treatment for aged MD patients may decrease the risk of AD, and agents that increase central BDNF may offer an alternative treatment for MD patients with cognitive impairment and/or for AD sufferers with depressive symptoms. PMID- 12781653 TI - Cell brain: a new focus in the postgenomic era. AB - The centrosome, together with the embedded centrioles and connecting filaments, has come to be regarded as the 'brain' of a cell, analogous to the long known brain of an animal or a human being. It is through the 'brain' that different cellular activities are coordinated as a whole. In this article, comparative studies of the principles of life at varying levels and of the new roles of different cellular organelles in maintaining a healthy life for an organism provide further support to this theory, which is discussed based on the latest findings. Hopefully, this new theory can make a great contribution to break the paradigm of nucleus (or genes) as causes of all problems. PMID- 12781654 TI - Cell brain abnormalities in cancer development. AB - Cancer has long been regarded as a genetic disease. Therefore, current theories on cancer development focus on genetic alterations affecting oncogenes or tumor suppressor genes. However, the mechanisms through which genetic alterations are induced are largely unknown. In this paper a theory will be developed which interprets cancer as a cell brain illness rather than a genetic disease. The complex comprising the centrosome, normally two centrioles and connecting filaments, was recently termed the 'cell brain', and was found to determine a cell's fate. It is through the cell brain, instead of the genes, that genetic stability and expression are maintained and regulated. Accordingly, the nucleus is regarded as a safe storage for inheriting materials (genes) that primarily act as manufacturing templates. Therefore, cancer should be regarded as a 'brain illness' of a cell, instead of a genetic disease, which is strongly supported by the latest evidence, as discussed in this paper. Such a theory serves to better clarify the confusing observations in cancer development accumulated over the last decades. PMID- 12781655 TI - The centrosome-centered cell-brain in apoptosis. AB - Apoptosis (or programmed cell death) is one of the central cellular processes in development, stress response, aging, carcinogenesis, and disease in multi cellular eukaryotes. Although great effort has been made, the detailed mechanism through which apoptosis is initiated is yet unclear. Previously, the centrosome, or more explicitly the complex comprising the centrosome, centrioles, and connecting filaments, was reported to be required for apoptosis. It may be through this 'cell brain', reminiscent of the long known brain of animals (or humans), that complicated cellular processes, including apoptosis, are precisely coordinated. In this paper, the latest data to support this contention are scrutinized. PMID- 12781656 TI - Does reduction of blood prolactin levels reveal the activation of central dopaminergic pathways conveying reward in top athletes? AB - Prolactin blood levels are modulated directly by CNS via the known tubero infundibular dopaminergic neurons which exert an inhibitory action on lactotrope cells in the hypophysis. Prolactin is devoid of peripheral negative feedback and via lesser known central pathways (e.g., mesolimbic and mesocortical) might be further tuned by means of appropriate stimuli (e.g., addictive drugs, gambling, exercise and so forth). Therefore, a prolactin test can be utilised to obtain an objective index to assess the compliance to exercise in men. This index may be used to identify the most suitable athlete for a given sport and mainly to survey exercise as rewarding behaviour in trained people. Prolactin was assessed in male top world windsurfers (WS) and in a control group of non-windsurfer trained subjects (CS) during an exercise simulating marine windsurfing. The result was that prolactin levels decreased significantly in WS with respect to CS when compared with pre-test levels (WS: -22.7%; P<0.05). PMID- 12781657 TI - HIV, 'an evolving species'. Roles of cellular activation and co-infections. AB - Each small variation of the genome of a species can be preserved if it is useful for the survival of the species in a given environment. Within this framework, the finality of the biological cycle of HIV consists in a search for harmony (biological coherence) with man, which is to say a stable condition. Cellular activation appears to be the strategy developed by HIV in order to achieve this coherence. The price of this strategy is the AIDS. The first contact between HIV and immune system appears to determine the subsequent clinical outcome and the future of HIV. Lymphocytic activation varies during the course of the vital cycle of HIV. For each individual, this lymphocytic activation depends on both the HLA repertoire acquired during thymic ontogenesis and the antigenic experience before and after HIV infection. Thus intercurrent infections alter the immune condition of the organism and influence the outcome of HIV. We described a synthetic analysis of the effects of HIV on the surface protein expression and the cellular activation pathways which should provide insights in the evolutionary relationship between HIV and man and should permit to do a more physiological therapeutic approach. PMID- 12781658 TI - More than synchrony: EEG chaoticity may be necessary for conscious brain functioning. AB - Transient phase synchronization of EEG oscillations was reported to be essential for conscious brain activity. However, it has been argued that EEG synchrony is not the only necessary factor, but at least must be accompanied by a sufficient complexity of brain dynamics. Here, we propose that, besides complexity, EEG chaoticity may be another necessary condition for conscious brain functioning. PMID- 12781659 TI - Microstructure, mechanical, and biomimetic properties of fish scales from Pagrus major. AB - The fish scale of Pagrus major has an orthogonal plywood structure of stratified lamellae, 1-2 microm in thickness, consisting of closely packed 70- to 80-nm diameter collagen fibers. X-ray diffraction, energy-dispersive X-ray analysis, and infrared spectroscopy indicate that the mineral phase in the scale is calcium deficient hydroxyapatite containing a small amount of sodium and magnesium ions, as well as carbonate anions in phosphate sites of the apatite lattice. The tensile strength of the scale is high (approximately 90 MPa) because of the hierarchically ordered structure of mineralized collagen fibers. Mechanical failure occurs by sliding of the lamellae and associated pulling out and fracture of the collagen fibers. In contrast, demineralized scales have significantly lower tensile strength (36 MPa), indicating that interactions between the apatite crystals and collagen fibers are of fundamental importance in determining the mechanical properties. Thermal treatment of fish scales to remove the organic components produces remarkable inorganic replicas of the native orthogonal plywood structure of the fibrillary plate. The biomimetic replica produced by heating to 873 K consists of stratified porous lamellae of c-axis-aligned apatite crystals that are long, narrow plates, 0.5-0.6 microm in length and 0.1-0.2 microm in width. The textured inorganic material remains intact when heated to 1473 K, although the size of the constituent crystals increases as a result of thermal sintering. PMID- 12781660 TI - Accurate determination of local defocus and specimen tilt in electron microscopy. AB - Accurate knowledge of defocus and tilt parameters is essential for the determination of three-dimensional protein structures at high resolution using electron microscopy. We present two computer programs, CTFFIND3 and CTFTILT, which determine defocus parameters from images of untilted specimens, as well as defocus and tilt parameters from images of tilted specimens, respectively. Both programs use a simple algorithm that fits the amplitude modulations visible in a power spectrum with a calculated contrast transfer function (CTF). The background present in the power spectrum is calculated using a low-pass filter. The background is then subtracted from the original power spectrum, allowing the fitting of only the oscillatory component of the CTF. CTFTILT determines specimen tilt parameters by measuring the defocus at a series of locations on the image while constraining them to a single plane. We tested the algorithm on images of two-dimensional crystals by comparing the results with those obtained using crystallographic methods. The images also contained contrast from carbon support film that added to the visibility of the CTF oscillations. The tests suggest that the fitting procedure is able to determine the image defocus with an error of about 10nm, whereas tilt axis and tilt angle are determined with an error of about 2 degrees and 1 degrees, respectively. Further tests were performed on images of single protein particles embedded in ice that were recorded from untilted or slightly tilted specimens. The visibility of the CTF oscillations from these images was reduced due to the lack of a carbon support film. Nevertheless, the test results suggest that the fitting procedure is able to determine image defocus and tilt angle with errors of about 100 nm and 6 degrees, respectively. PMID- 12781661 TI - The sperm roller: a modified testicular duct linked to giant sperm transport within the male reproductive tract. AB - The male reproductive tracts of Drosophila display considerable variation in the relative size of their components, notably of the testes, but there are few structural differences between species. Here we report a remarkable coiled structure separating the testes from the seminal vesicles in the giant sperm species Drosophila bifurca. This evolutionary novelty, known as the 'sperm roller', seems to be an exaggeration in the size of the testicular duct as revealed by light and electron microscopic observations. It consists of a tubular monocellular epithelium lying on the basal laminae and muscle and conjunctive cells. The lumen of the roller contains crypts. The apical membrane of the epithelial cells presents numerous long microvilli protruding into the lumen. The sperm roller structure is probably involved in managing sperm during their transit through the male genital tract, because sperm are seen in bundles at the base of the testis, whereas they are singly rolled up by the time they enter the seminal vesicles. The hypercoiling of the individual spermatozoon within the roller probably occurs as the result of an osmotic process produced by features of the epithelial wall and the dramatically increased exchange surface. This is the first report of a specialized device of this type in Drosophila or, more generally, in insects. PMID- 12781662 TI - Pseudo-atomic models of swollen CCMV from cryo-electron microscopy data. AB - The capsid of cowpea chlorotic mottle virus (CCMV) can reversibly switch between two forms that are contingent on the charge of acidic residues that are clustered at the quasi-threefold axes of the T=3 icosahedral particle. The quaternary structure conformations are dependent on divalent metal ions and pH and were previously analyzed by crystallography in the native, compact form, and by cryo electron microscopy in the compact and swollen forms (Speir et al., 1995). In this report we use the atomic models of the three structurally unique viral subunits determined by crystallography for a detailed interpretation of the 28-A resolution electron density of the swollen form and the production of a pseudo atomic model of this particle. The model of the quaternary structure conforms with high fidelity to conventional geometric constraints, quasi-equivalence, intersubunit association energies, and the electron density. It was derived by conserving the pentamers and hexamers of subunits whose associated electron densities are strikingly similar in the two forms of the particles. Treating these as rigid units in the modeling implies that the particle flexibility is accommodated primarily by changes in dimer interactions, an observation that is consistent with the flexible C-terminal polypeptide extensions that stabilize this contact in the crystal structure. Because the hexamers and pentamers were incrementally translated and rotated in a screw motion, with energy minimization at each of 28 steps, a path for the expansion is also implied. PMID- 12781663 TI - Raman-atomic force microscopy of the ommatidial surfaces of Dipteran compound eyes. AB - The ommatidial lens surfaces of the compound eyes in several species of files (Insecta: Diptera) and a related order (Mecoptera) were analyzed using a recently developed Raman-atomic force microscope. We demonstrate in this work that the atomic force microscope (AFM) is a potentially useful instrument for gathering phylogenetic data and that the newly developed Raman-AFM may extend this application by revealing nanometer-scale surface chemistry. This is the first demonstration of apertureless near-field Raman spectroscopy on an intact biological surface. For Chrysopilus testaceipes Bigot (Rhagionidae), this reveals unique cerebral cortex-like surface ridges with periodic variation in height and surface chemistry. Most other Brachyceran flies, and the "Nematoceran" Sylvicola fenestralis (Scopoli) (Anisopodidae), displayed the same morphology, while other taxa displayed various other characteristics, such as a nodule-like (Tipula (Triplicitipula) sp. (Tipulidae)) or coalescing nodule-like (Tabanus punctifer Osten Sacken (Tabanidae)) morphology, a smooth morphology with distinct pits and grooves (Dilophus orbatus (Say) (Bibionidae)), or an entirely smooth surface (Bittacus chlorostigma MacLachlan (Mecoptera: Bittacidae)). The variation in submicrometer structure and surface chemistry provides a new information source of potential phylogenetic importance, suggesting the Raman-atomic force microscope could provide a new tool useful to systematic and evolutionary inquiry. PMID- 12781664 TI - Determining molecular forces that stabilize human aquaporin-1. AB - Atomic force microscopy (AFM) was used to measure the forces stabilizing human aquaporin-1 (hAQP1), a tetrameric transmembrane protein that forms highly specific water channels. To this end, the AFM tip was attached to the C-terminus of hAQP1 and secondary structure elements were extracted from the membrane while the single-molecule force-extension curve was being recorded. Force peaks, reflecting the unfolding of secondary structure elements, could be interpreted in depth using the atomic model of hAQP1. Different classes of force-extension curves indicated the existence of alternative unfolding pathways for individual proteins. In addition, transmembrane helices at the periphery of the hAQP1 tetramer exhibited smaller extraction forces than helices at the interface between hAQP1 monomers. These results represent the first direct assessment of intermolecular forces defining the oligomeric state of a membrane protein. PMID- 12781665 TI - Hsp70 and Hsp90 change their expression and subcellular localization after microspore embryogenesis induction in Brassica napus L. AB - A stress treatment of 32 degrees C for at least 8h was able to change the gametophytic program of the microspore, switching it to embryogenesis in Brassica napus, an interesting model for studying this process in vitro. After induction, some microspores started symmetric divisions and became haploid embryos after a few days, whereas other microspores, not sensitive to induction, followed their original gametophytic development. In this work the distribution and ultrastructural localization of two heat-shock proteins (Hsp70 and Hsp90) throughout key stages before and after embryogenesis induction were studied. Both Hsp proteins are rapidly induced, localizing in the nucleus and the cytoplasm. Immunogold labeling showed changes in the distribution patterns of these proteins, these changes being assessed by a quantitative analysis. Inside the nucleus, Hsp70 was found in association with RNP structures in the interchromatin region and in the nucleolus, whereas nuclear Hsp90 was mostly found in the interchromatin region. For Hsp70, the accumulation after the inductive treatment was accompanied by a reversible translocation from the cytoplasm to the nucleus, in both induced (embryogenic) and noninduced (gametophytic) microspores. However, the translocation was higher in embryogenic microspores, suggesting a possible additional role for Hsp70 in the switch to embryogenesis. In contrast, Hsp90 increase was similar in all microspores, occurring faster than for Hsp70 and suggesting a more specific role for Hsp90 in the stress response. Hsp70 and Hsp90 colocalized in clusters in the cytoplasm and the nucleus, but not in the nucleolus. Results indicated that stress proteins are involved in the process of microspore embryogenesis induction. The differential appearance and distribution of the two proteins and their association at specific stages have been determined between the two systems coexisting in the same culture: embryogenic development (induced cells) and development of gametes (noninduced cells). PMID- 12781666 TI - Automatic CTF correction for single particles based upon multivariate statistical analysis of individual power spectra. AB - Three-dimensional electron cryomicroscopy of randomly oriented single particles is a method that is suitable for the determination of three-dimensional structures of macromolecular complexes at molecular resolution. However, the electron-microscopical projection images are modulated by a contrast transfer function (CTF) that prevents the calculation of three-dimensional reconstructions of biological complexes at high resolution from uncorrected images. We describe here an automated method for the accurate determination and correction of the CTF parameters defocus, twofold astigmatism and amplitude-contrast proportion from single-particle images. At the same time, the method allows the frequency dependent signal decrease (B factor) and the non-convoluted background signal to be estimated. The method involves the classification of the power spectra of single-particle images into groups with similar CTF parameters; this is done by multivariate statistical analysis (MSA) and hierarchically ascending classification (HAC). Averaging over several power spectra generates class averages with enhanced signal-to-noise ratios. The correct CTF parameters can be deduced from these class averages by applying an iterative correlation procedure with theoretical CTF functions; they are then used to correct the raw images. Furthermore, the method enables the tilt axis of the sample holder to be determined and allows the elimination of individual poor-quality images that show high drift or charging effects. PMID- 12781667 TI - Aprotinin uptake in the proximal tubules in the rat kidney I. Length of proximal tubular uptake segment. AB - Aprotinin (Ap), a basic polypeptide with a molecular weight of 6500, is filtered at the glomerular membrane without steric restriction and is completely absorbed by the proximal tubule cells. Here Ap is broken down to amino acids, but no breakdown products enter the peritubular circulation during the first 20 min following an intravenous injection. These properties have recently been exploited for measurement of local glomerular filtration rate, based on the assumption that the proximal tubular uptake site is located at the level of the filtering glomerulus. To evaluate that assumption we have now made serial autoradiographs of the rat kidney 20 min after intravenous injection of 2-750 microg of 125I Aprotinin. With all doses the percent 125I-containing proximal tubular transections were about 50 in the outer and middle cortex and 35 in the inner third. We interpret these numbers to mean that all filtered Ap is taken up in the first two thirds of the proximal convoluted tubular length and does not reach the pars recta. Since the proximal tubule on average is located more superficial than its glomerulus, measurement of local Ap uptake will tend to overestimate glomerular filtration rate in outer layers of the cortex. Quantitative estimate of this "displacement" will be presented in a companion article. PMID- 12781668 TI - Aprotinin uptake in the proximal tubules in the rat kidney. II. Uptake site relative to glomerulus. AB - Glomerular filtration rates in whole kidney and in outer, middle and inner cortical zones have previously been estimated by measuring the amount of iodinated Aprotinin, filtered and taken up in the first two thirds of the proximal convoluted tubules, in part positioned more superficial than the parent glomerulus. Thus, an appreciable amount of the absorbed Aprotinin may be located superficial to its filtration site and lead to an underestimate of glomerular filtration in deep cortical layers. Therefore, in this study we have measured the distance from the glomerulus to the center of proximal convoluted tubular ball and the site of Aprotinin uptake. Measurements were made on photos of Microfil injected tubules and on camera lucida drawings of tubular transections from autoradiographs of nephrons containing both Microfil and iodinated Aprotinin. Both techniques showed that the center of the tubular ball was localized more superficial in all cortical layers. The average distance, in percent of cortical thickness, from all proximal convoluted tubular transections to the parent glomerulus was 9% in deep and 13% in middle and superficial cortex. Corresponding distances for tubular transections containing Aprotinin were 7 and 12%. Grain density in five reconstructed proximal convoluted tubules showed a continuous and exponential fall of Aprotinin along the uptake segment. The results may be used to estimate single nephron filtration rate from Aprotinin uptake and glomerular density in outer, middle, and inner cortex. PMID- 12781669 TI - Mycotoxin production by indoor molds. AB - Fungal growth in buildings starts at a water activity (a(w)) near 0.8, but significant quantities of mycotoxins are not produced unless a(w) reaches 0.95. Stachybotrys generates particularly high quantities of many chemically distinct metabolites in water-damaged buildings. These metabolites are carried by spores, and can be detected in air samples at high spore concentrations. Very little attention has been paid to major metabolites of Stachybotrys called spirocyclic drimanes, and the precise structures of the most abundant of these compounds are unknown. Species of Aspergillus and Penicillium prevalent in the indoor environment produce relatively low concentrations of mycotoxins, with the exception of sterigmatocystins that can represent up to 1% of the biomass of A. versicolor at a(w)'s close to 1. The worst-case scenario for homeowners is produced by consecutive episodes of water damage that promote fungal growth and mycotoxin synthesis, followed by drier conditions that facilitate the liberation of spores and hyphal fragments. PMID- 12781670 TI - Identification of genes differentially expressed during aflatoxin biosynthesis in Aspergillus flavus and Aspergillus parasiticus. AB - A complex regulatory network governs the biosynthesis of aflatoxin. While several genes involved in aflatoxin production are known, their action alone cannot account for its regulation. Arrays of clones from an Aspergillus flavus cDNA library and glass slide microarrays of ESTs were screened to identify additional genes. An initial screen of the cDNA clone arrays lead to the identification of 753 unique ESTs. Many showed sequence similarity to known metabolic and regulatory genes; however, no function could be ascribed to over 50% of the ESTs. Gene expression analysis of Aspergillus parasiticus grown under conditions conducive and non-conductive for aflatoxin production was evaluated using glass slide microarrays containing the 753 ESTs. Twenty-four genes were more highly expressed during aflatoxin biosynthesis and 18 genes were more highly expressed prior to aflatoxin biosynthesis. No predicted function could be ascribed to 18 of the 24 genes whose elevated expression was associated with aflatoxin biosynthesis. PMID- 12781671 TI - Deletion of a lectin gene does not affect the phenotype of the nematode-trapping fungus Arthrobotrys oligospora. AB - A number of filamentous fungi are known to produce high levels of saline-soluble and low-molecular-mass lectins. The function of these proteins are not clear but it has been proposed that they are involved in storage of nutrients, development, recognition of other organisms, and defense reactions. A gene encoding such a lectin (AOL) was deleted in the nematode-trapping fungus Arthrobotrys oligospora by homologous recombination. The deletion mutants did not express any hemagglutinating activity or protein cross-reacting with AOL antibodies. There were no significant differences between the DeltaAOL and wild-type strains in spore (conidia) germination, saprophytic growth, and pathogenicity. Furthermore, there was no significant difference in the growth and reproduction of collembolan feeding on the various strains of A. oligospora. Thus either the previous proposed functions of AOL are not correct, or the fungus can compensate for the absence of the lectin by expressing other proteins with similar function(s) as AOL. PMID- 12781672 TI - Male and female roles in crosses of Aspergillus nidulans as revealed by vegetatively incompatible parents. AB - To resolve the role of male and female nuclei and mitochondria in cleistothecium formation in the model organism Aspergillus nidulans, we analysed the genetic constituents of cleistothecia from crosses between vegetatively compatible and incompatible parents. We used markers that enabled us to determine the nuclear genotype of the cleistothecial wall and the nuclear and mitochondrial genotype of the ascospores. In compatible parents, nuclear genomes and cytoplasm usually mix in the vegetative hyphae prior to the formation of the sexual stage after which any cleistothecial composition is possible. In incompatible parents, the maternal strain contributes the nuclei for the cleistothecial wall and one nucleus as well as mitochondria for the ascospore origin. The paternal strain donates one nucleus for the ascospore origin. Only in crosses between vegetatively incompatible partners, it is possible to assign a female and male role to the parental strains. Our results confirm that the vegetative heterokaryotic stage is not a prerequisite for cleistothecium formation. Using this tool, we analysed sexual sporulation mutants for male or female sterility. PMID- 12781673 TI - Analysis of two additional loci in Neurospora crassa related to Spore killer-2. AB - Two new loci found in one strain of Neurospora crassa (P2604) collected in Malaya are related to the meiotic drive system Spore killer Sk-2. Sk-2 was found in Neurospora intermedia and introgressed into N. crassa. P2604 showed high resistance to killing when crossed to Sk-2. This resistance was found to be linked to, but not allelic to, resistance locus r(Sk-2) on LGIIIL. Analysis showed that the high resistance phenotype of P2604 requires resistance alleles at two different loci on LGIIIR. Strains carrying a resistance allele at only the proximal or the distal locus, respectively, were obtained and intercrossed. Highly resistant strains were obtained by rejoining the two genes. The proximal locus alone confers a low level of resistance. This locus was named pr(Sk-2) for partial resistance to Sk-2. The distal locus was named mod(pr) because its only known phenotype is to modify pr(Sk-2). PMID- 12781674 TI - Cloning and characterization of the mating type (MAT) locus from Ascochyta rabiei (teleomorph: Didymella rabiei) and a MAT phylogeny of legume-associated Ascochyta spp. AB - Degenerate primers designed to correspond to conserved regions of the high mobility group (HMG) protein encoded by the MAT1-2 gene of Cochliobolus heterostrophus, Cochliobolus sativus, and Alternaria alternata were used to amplify the portion of the sequence corresponding to the HMG box motif from Ascochyta rabiei (teleomorph: Didymella rabiei). A combination of TAIL and inverse PCR extended the MAT1-2 sequence in both directions, then primers designed to MAT1-2 flanking DNA were used to amplify the entire MAT1-1 idiomorph. MAT1-1 and MAT1-2 idiomorphs were 2294 and 2693 bp in length, respectively, and each contained a single putative open reading frame (ORF) and intron similar to MAT loci of other loculoascomycete fungi. MAT genes were expressed at high levels in rich medium. MAT-specific PCR primers were designed for use in a multiplex PCR assay and MAT-specific PCR amplicons correlated perfectly to mating phenotype of 35 ascospore progeny from a cross of MAT1-1 by MAT1-2 isolates and to the mating phenotype of field-collected isolates from diverse geographic locations. MAT specific PCR was used to rapidly determine the mating type of isolates of A. rabiei sampled from chickpea fields in the US Pacific Northwest. Mating type ratios were not significantly different from 1:1 among isolates sampled from two commercial chickpea fields consistent with the hypothesis that these A. rabiei populations were randomly mating. The mating type ratio among isolates sampled from an experimental chickpea field where asexual reproduction was enforced differed significantly from 1:1. A phylogeny estimated among legume-associated Ascochyta spp. and related loculoascocmycete fungi using sequence data from the nuclear ribosomal internal transcribed spacer (ITS) demonstrated the monophyly of Ascochyta/Didymella spp. associated with legumes but was insufficiently variable to differentiate isolates associated with different legume hosts. In contrast, sequences of the HMG region of MAT1-2 were substantially more variable, revealing seven well-supported clades that correlated to host of isolation. A. rabiei on chickpea is phylogenetically distant from other legume-associated Ascochyta spp. and the specific status of A. rabiei, A. lentis, A. pisi, and A. fabae was confirmed by the HMG phylogeny PMID- 12781675 TI - Carbohydrate and amino acid metabolism in Tuber borchii mycelium during glucose utilization: a (13)C NMR study. AB - The metabolism of [1-13C]glucose in the vegetative mycelium of the ectomycorrhizal ascomycete Tuber borchii was studied in order to characterize the biochemical pathways for the assimilation of glucose and amino acid biosynthesis. The pathways were characterized using nuclear magnetic resonance spectroscopy in conjunction with [1-13C]glucose labeling. The enzymes of mannitol cycle and ammonium assimilation were also evaluated. The majority of the 13C label was incorporated into mannitol and this polyol was formed via a direct route from absorbed glucose. Amino acid biosynthesis was also an important sink of assimilated carbon and 13C was mainly incorporated into alanine and glutamate. From this intramolecular 13C enrichment, it is concluded that pyruvate, arising from [1-13C]glucose catabolism, was used by alanine aminotransferase, pyruvate dehydrogenase and pyruvate carboxylase before entering the Krebs cycle. The transfer of 13C-labeled mycelium on [12C]glucose showed that mannitol, alanine, and glutamate carbon were used to synthesize glutamine and arginine that likely play a storage role. PMID- 12781676 TI - Alternative oxidase expression in Neurospora crassa. AB - When electron flow through the cytochrome-mediated electron transport chain is blocked by inhibitors or mutations, the mitochondria of Neurospora crassa contain a KCN-insensitive alternative oxidase, encoded by the aod-1 gene, that transfers electrons directly from the ubiquinone pool to oxygen. The mechanism by which the enzyme is induced is unknown. Comparison of the sequence upstream of the N. crassa aod-1 gene with the corresponding region of Gelasinospora spp. and Aspergillus nidulans revealed a cyclic AMP responsive element (CRE) about 700-800 bp upstream of the start codon in each species. Electrophoretic mobility shift assays showed that a factor from N. crassa cell extracts binds specifically to the CRE sequence. However, transformation of an aod-1 mutant strain with constructs lacking the CRE gave strains that regulate alternative oxidase in a normal fashion. Nuclear run-on assays indicated that uninduced cells transcribe the aod-1 gene at a low constitutive rate and that the transcription rate is increased in cells induced by antimycin A. Non-induced wild-type cultures occasionally contained significant amounts of aod-1 mRNA, but Western blots revealed no detectable AOD1 protein in mitochondria of these cells. This suggests that post-transcriptional events also play a role in alternative oxidase expression. A BLAST search of the Neurospora genome sequence revealed a second gene with the potential to encode an alternative oxidase, which we have named aod 3. Northern blot analysis using probes specific for the aod-1 and aod-3 genes revealed no evidence of expression of aod-3. PMID- 12781677 TI - ESTs from the basidiomycete Schizophyllum commune grown on nitrogen-replete and nitrogen-limited media. AB - Lambda phage cDNA libraries were constructed using mRNAs from the basidiomycete Schizophyllum commune grown on media with high or low nitrogen concentrations. A total of 440 clones were sequenced, representing 373 distinct transcripts. Of these, 166 showed significant similarity to annotated genes in GenBank. Those that could be tentatively identified using BLAST searches were classified by function using the Gene Ontology (GO) database. Genes with products involved in cell-cycle processes were more frequent in the nitrogen-limited libraries, while genes with products involved in protein biosynthesis were more frequent in the nitrogen-replete library. Overall, clones showed much greater similarity to the one publicly available basidiomycete genome, Phanerochaete chrysosporium, than to any of the ascomycete genomes. PMID- 12781678 TI - Heart development: molecular insights into cardiac specification and early morphogenesis. AB - The heart develops from two bilateral heart fields that are formed during early gastrulation. In recent years, signaling pathways that specify cardiac mesoderm have been extensively analyzed. In addition, a battery of transcription factors that regulate different aspects of cardiac morphogenesis and cytodifferentiation have been identified and characterized in model organisms. At the anterior pole, a secondary heart field is formed, which in its molecular make-up, appears to be similar to the primary heart field. The cardiac outflow tract and the right ventricle to a large extent are derivatives of this anterior heart field. Cardiac mesoderm receives positional information by which it is patterned along the three body axes. The molecular control of left-right axis development has received particular attention, and the underlying regulatory network begins to emerge. Cardiac chamber development involves the activation of a transcription program that is different from the one present in the primary heart field and regulates cardiac morphogenesis in a region-specific manner. This review also attempts to identify areas in which additional research is needed to fully understand early cardiac development. PMID- 12781679 TI - Progressive restriction of cell fates in relation to neuroepithelial cell mingling in the mouse cerebellum. AB - Neurogenesis in the cerebellum proceeds through a temporal series of cell production from two separate epithelia, the ventricular zone (VZ) and the external granule cell layer (EGL). Using the laacZ cell lineage tracer in transgenic mice, we describe cellular clones whose dates of birth span the entire period of cerebellar development and deduce a sequence of cell dispersion leading to the final allocation of cells in the cerebellum. Clones probably labeled early during neural tube formation show that individual progenitors can give rise to all cerebellar cell types. The distribution of clonally related granule cells in these clones indicates a mediolateral organization of EGL progenitors already established before the allocation of the EGL progenitors to the cerebellum. Clones restricted to the cerebellar VZ show that the VZ derives progenitors for deep nuclei and multipotent cortical progenitors, which lose their systematic lineage relationship when longitudinal cell intermingling in the cerebellar VZ becomes more limited. The small clones also show that cell dispersion is radial in the internal granule layer and tangential in the molecular layer. Together, the data demonstrate the broad maintenance of the relative order of cells from neural tube stages to the adult cerebellum. PMID- 12781681 TI - Evolution of identified arthropod neurons: the serotonergic system in relation to engrailed-expressing cells in the embryonic ventral nerve cord of the american lobster homarus americanus milne edwards, 1873 (malacostraca, pleocyemata, homarida). AB - One of the long-standing questions in zoology is that on the phylogenetic relationships within the Arthropoda. Comparative studies on structure and development of the nervous system can contribute important arguments to this discussion. In the present report, the arrangement of serotonin- and engrailed expressing cells was examined in the embryonic ventral nerve cord of the American lobster Homarus americanus Milne Edwards, 1873 (Malacostraca, Pleocyemata, Homarida), and the spatial relationship of these two cell classes was explored by a double-labelling approach. The goal of this study was to determine whether the lobster serotonergic neurons are homologous to similar cells present in representatives of the Hexapoda and other Arthropoda. The results indicate that, in fact, these neurons in the lobster ventral nerve cord have corresponding counterparts in many other mandibulate taxa. Based on the finding of these homologies, the arrangement of serotonergic neurons in a model trunk ganglion of the mandibulate ground pattern was reconstructed as comprising an anterior and a posterior pair of serotonergic neurons per hemiganglion, each cell with both an ipsilateral and a contralateral neurite. Starting from this ground pattern, the evolutionary diversification of this class of neurons within the Mandibulata is discussed. PMID- 12781680 TI - Activation of pmar1 controls specification of micromeres in the sea urchin embryo. AB - pmar1 is a transcription factor in the paired class homeodomain family that was identified and found to be transcribed in micromeres beginning at the fourth cleavage of sea urchin development [Dev. Biol. 246 (2002), 209]. Based on in situ data, molecular perturbation studies, and QPCR data, the recently published gene regulatory network (GRN) model for endomesoderm specification [Science 295 (2002) 1669; Dev. Biol. 246 (2002), 162] places pmar1 early in the micromere specification pathway, and upstream of two important micromere induction signals. The goal of this study was to test these three predictions of the network model. A series of embryo chimeras were produced in which pmar1 activity was perturbed in one cell that was transplanted to control hosts. At the fourth cleavage, micromeres bearing altered pmar1 activity were combined with a normal micromereless host embryo. If beta-catenin signaling is blocked, the micromeres remain unspecified and are unable to signal to the host cells. When such beta catenin-blocked micromeres also express Pmar1, all observed micromere functions are rescued. The rescue includes expression of the primary mesenchyme cell (PMC) differentiation program, expression and execution of the Delta signal to induce secondary mesoderm cell (SMC) specification in macromere progeny, and expression of the early endomesoderm induction signal necessary for full specification of the endoderm. Additionally, Pmar1 expressed mosaically from inserted DNA constructs causes induction of ectopic Endo 16 in adjacent cells, demonstrating further that Pmar1 controls expression of the early endomesoderm induction signal. Based on these experiments, Pmar1 is an important transcription factor necessary for initiating the micromere specification program and for the expression of two inductive signals produced by micromeres. Each of the tests we describe supports the placement and function of Pmar1 in the endomesoderm GRN model. PMID- 12781682 TI - Embryonic cell lineage of the marine nematode Pellioditis marina. AB - We describe the complete embryonic cell lineage of the marine nematode Pellioditis marina (Rhabditidae) up to somatic muscle contraction, resulting in the formation of 638 cells, of which 67 undergo programmed cell death. In comparison with Caenorhabditis elegans, the overall lineage homology is 95.5%; fate homology, however, is only 76.4%. The majority of the differences in fate homology concern nervous, epidermal, and pharyngeal tissues. Gut and, remarkably, somatic muscle is highly conserved in number and position. Partial lineage data from the slower developing Halicephalobus sp. (Panagrolaimidae) reveal a lineage largely, but not exclusively, built up of monoclonal sublineage blocs with identical fates, unlike the polyclonal fate distribution in C. elegans and P. marina. The fate distribution pattern in a cell lineage could be a compromise between minimizing the number of specification events by monoclonal specification and minimizing the need for migrations by forming the cells close at their final position. The latter could contribute to a faster embryonic development. These results reveal that there is more than one way to build a nematode. PMID- 12781683 TI - Contrasting patterns of mitochondrial redistribution in the early lineages of Caenorhabditis elegans and Acrobeloides sp. PS1146. AB - We compared the redistribution of mitochondria in the early embryos of Caenorhabditis elegans (C. elegans) and Acrobeloides sp. PS1146 (Acrobeloides)- two nematode species where the mechanisms for embryonic axis specification are different even though subsequent development is remarkably similar. During the first cell cycle of C. elegans, mitochondria move with the bulk cytoplasmic flows that are directed toward the sperm pronucleus and aggregate at the posterior cortex during the period known as "pseudocleavage." In contrast, in Acrobeloides embryos, where prominent cytoplasmic rearrangements are absent, mitochondria that are initially distributed loosely around the pronuclei and the cytoplasm are relocated around the mitotic spindle prior to cell division. Interestingly, this rearrangement is reiterated only in the germline and not the somatic lineage. In both species, the location of the mitochondria immediately prior to cell division correlates with the known location of the germline determinants, P granules, leading us to speculate that they may be associated. PMID- 12781684 TI - E74 exhibits stage-specific hormonal regulation in the epidermis of the tobacco hornworm, manduca sexta. AB - The transcription factor E74 is one of the early genes induced by ecdysteroids during metamorphosis of Drosophila melanogaster. Here, we report the cloning and hormonal regulation of E74 from the tobacco hornworm, Manduca sexta (MsE74). MsE74 is 98% identical to that of D. melanogaster within the DNA-binding ETS domain of the protein. The 5'-isoform-specific regions of MsE74A and MsE74B share significantly lower sequence similarity (30-40%). Developmental expression by Northern blot analysis reveals that, during the 5th larval instar, MsE74B expression correlates with pupal commitment on day 3 and is induced to maximal levels within 12h by low levels of 20-hydroxyecdysone (20E) and repressed by physiologically relevant levels of juvenile hormone I (JH I). Immunocytochemical analysis shows that MsE74B appears in the epidermis before the 20E-induced Broad transcription factor that is correlated with pupal commitment (Zhou and Riddiford, 2001). In contrast, MsE74A is expressed late in the larval and the pupal molts when the ecdysteroid titer has declined to low levels and in the adult molt just as the ecdysteroid titer begins to decline. This change in timing during the adult molt appears not to be due to the absence of JH as there was no change during the pupal molt of allatectomized animals. When either 4th or 5th instar larval epidermis was explanted and subjected to hormonal manipulations, MsE74A induction occurred only after exposure to 20E followed by its removal. Thus, MsE74B appears to have a similar role at the onset of metamorphosis in Manduca as it does in Drosophila, whereas MsE74A is regulated differently at pupation in Manduca than at pupariation in Drosophila. PMID- 12781685 TI - Microarray analysis of somitogenesis reveals novel targets of different WNT signaling pathways in the somitic mesoderm. AB - WNT signaling plays a major role in patterning the dermomyotome of the somitic mesoderm. However, knowledge of downstream target genes and their regulation is limited. To identify new genes involved in the development and early patterning of the somite, we performed a comparison of gene expression by microarray between the presomitic mesoderm and the 5 most recently formed somites of the mouse at embryonic day 9.5. We identified 207 genes upregulated and 120 genes downregulated in somite formation. Expression analysis and functional categorization of these genes demonstrate this to be a diverse pool that provides a valuable resource for studying somite development. Thus far, we have found three genes expressed in the dermomyotome of the early somite. Consistent with their expression patterns, these genes are transcriptional targets of WNT signals, but display differential activation by different WNTs. We further demonstrate that 1 of these genes, Troy, is a direct target of canonical WNT signaling, while the other 2 genes, Selp and Arl4, are not. Thus, our microarray study using microdissected tissues not only provides global information on gene expression during somite development, it also provides novel targets to study the inductive signaling pathways that direct somite patterning. PMID- 12781686 TI - The Onecut transcription factor HNF-6 (OC-1) is required for timely specification of the pancreas and acts upstream of Pdx-1 in the specification cascade. AB - The pancreas derives from cells in the ventral and dorsal foregut endoderm that express the transcription factor Pdx-1. These specified cells give rise to the precursors of the endocrine, ductal, and exocrine pancreatic cells. The identification of transcription factors that regulate the onset of Pdx-1 expression is therefore essential to understand pancreas development. No such factor that acts both in the ventral and in the dorsal endoderm is known. We showed previously that the Onecut transcription factor HNF-6 promotes differentiation of the endocrine cell precursors in which it stimulates expression of the proendocrine gene Ngn-3. By analyzing the phenotype of HNF-6 null mice, we now demonstrate that HNF-6 also controls an earlier step in pancreas development. Indeed, the pancreas of Hnf6(-/-) mice was hypoplastic. This did not result from decreased proliferation or from increased apoptosis, but from retarded pancreatic specification of endodermal cells. The onset of Pdx-1 expression was delayed both in the ventral and in the dorsal endoderm, leading to a reduction in the number of endodermal cells expressing Pdx-1 at the time of pancreatic budding. In normal embryos, HNF-6 was detected in the endoderm prior to the expression of Pdx-1. Moreover, HNF-6 could directly stimulate the Pdx1 promoter. Our data therefore identify HNF-6 as the first factor known to control Pdx-1 expression both in the ventral and in the dorsal endoderm. We conclude that HNF-6 controls the timing of pancreas specification and that HNF-6 acts upstream of Pdx-1 in this developmental process. Together with the known role of HNF-6 in pancreatic endocrine cell differentiation, our data point to HNF-6 as a key regulator of pancreas development. PMID- 12781687 TI - A somatic role for eyes absent (eya) and sine oculis (so) in Drosophila spermatocyte development. AB - Interactions between the soma and the germline are a conserved feature of spermatogenesis throughout the animal kingdom. In this report, we find that the transcription factors eyes absent (eya) and sine oculis (so), previously shown to play major roles during eye development [Cell 91 (1997), 881] are each required in the somatic cyst cells of the testis for proper Drosophila spermatocyte development. eya mutant testes exhibit degenerating young spermatocytes. Mosaic analysis reveals a somatic requirement for both eya and so, in that neither gene is required in the germline for spermatocyte development. Immunolocalization analysis supports this somatic role, since both proteins are localized within cyst cell nuclei as spermatocytes differentiate from amplifying spermatogonia. Using antibodies against known cyst cell markers, we demonstrate that cysts of degenerating spermatocytes in eya mutant testes are encysted, ruling out a role for eya in cyst cell viability. Finally, we have uncovered a genetic interaction between eya and so in the testis, suggesting that, as in the eye, eya and so may form a transcription complex responsible for the activation of target genes involved in cyst cell differentiation and spermatocyte development. PMID- 12781688 TI - Interaction between noradrenaline or adrenaline and the beta 1-adrenergic receptor in the nervous system triggers early metamorphosis of larvae in the ascidian, Ciona savignyi. AB - Molecular mechanisms underlying the metamorphosis of larvae, e.g., ligand and receptor interaction, have to be determined and roles for the nervous system in marine invertebrates are not well understood. We report here that treatment of swimming larvae of the ascidian Ciona savignyi with noradrenaline or adrenaline promoted morphological changes in early metamorphosis, e.g., tail resorption. Antagonists of the beta-adrenergic receptor, propranolol, and the beta(1) adrenergic receptor, metoprolol, inhibited the noradrenaline-induced tail resorption, while an antagonist of the alpha-adrenergic receptor, phentolamine, and of the beta(2)- adrenergic receptor, butoxamine, had no inhibitory effects. In addition, a selective agonist of the beta-adrenergic receptor, isoproterenol, the concentration of which was lower than the effective concentration of the neurotransmitters, facilitated tail resorption. Immunohistochemical studies, using an anti-dopamine-hydroxylase antibody, showed that neurotransmitters such as noradrenaline and adrenaline localized around the brain vesicle of the larvae during metamorphosis. The beta(1)-adrenergic receptor stained with antibodies was localized on the nervous system. Temporal expression of the beta(1)-adrenergic receptor was intense in the nervous system in the larvae competent for metamorphosis. We propose that interactions between noradrenaline or adrenaline and the beta(1)-adrenergic receptor in the nervous system mediate the process of metamorphosis of Ciona larvae. PMID- 12781689 TI - Control of epidermal stem cell clusters by Notch-mediated lateral induction. AB - Stem cells in the basal layer of human interfollicular epidermis form clusters that can be reconstituted in vitro. In order to supply the interfollicular epidermis with differentiated cells, the size of these clusters must be controlled. Evidence suggests that control is regulated via differentiation of stem cells on the periphery of the clusters. Moreover, there is growing evidence that this regulation is mediated by the Notch signalling pathway. In this paper, we develop theoretical arguments, in conjunction with computer simulations of a model of the basal layer, to show that regulation of differentiation is the most likely mechanism for cluster control. In addition, we show that stem cells must adhere more strongly to each other than they do to differentiated cells. Developing our model further we show that lateral-induction, mediated by the Notch signalling pathway, is a natural mechanism for cluster control. It can not only indicate to cells the size of the cluster they are in and their position within it, but it can also control the cluster size. This can only be achieved by postulating a secondary, cluster wide, differentiation signal, and cells with high Delta expression being deaf to this signal. PMID- 12781690 TI - Temporal effects of Sprouty on lung morphogenesis. AB - Paracrine signaling mediated by FGF-10 and the FGF-R2IIIb receptor is required for formation of the lung. To determine the temporal requirements for FGF signaling during pulmonary morphogenesis, Sprouty-4 (Spry-4), an intracellular FGF receptor antagonist, was expressed in epithelial cells of the fetal lung under control of a doxycycline-inducible system. Severe defects in lobulation and severe lung hypoplasia were observed when Spry-4 was expressed throughout fetal lung development (E6.5-E18.5) or from E6.5 until E13.5. Effects of Spry-4 on branching were substantially reversed by removal of doxycycline from the dam at E12.5, but not at E13.5. In contrast, when initiated late in development (E12.5 to birth), Spry-4 caused less severe pulmonary hypoplasia. Expression of Spry-4 from E16.5 to E18.5 reduced lung growth and resulted in perinatal death due to respiratory failure. Expression of Spry-4 during the saccular and alveolar stages, from E18.5 to postnatal day 21, caused mild emphysema. These findings demonstrate that the embryonic-pseudoglandular stage is a critical time period during which Spry-sensitive pathways are required for branching morphogenesis, lobulation, and formation of the peripheral lung parenchyma. PMID- 12781691 TI - Tissue interactions pattern the mesenchyme of the embryonic mouse lung. AB - The mechanisms that control proliferation and differentiation of embryonic lung mesenchyme are largely unknown. We describe an explant system in which exogenous recombinant N-Sonic Hedgehog (N-Shh) protein sustains the survival and proliferation of lung mesenchyme in a dose-dependent manner. In addition, Shh upregulates several mesenchymal cell markers, including its target gene Patched (Ptc), intercellular signaling genes Bone Morphogenetic Protein-4 (Bmp4) and Noggin (Nog), and smooth muscle actin and myosin. In explants exposed to N-Shh in the medium, these products are upregulated throughout the mesenchyme, but not in the periphery. This exclusion zone correlates with the presence of an overlying mesothelial layer, which, as in vivo, expresses Fibroblast Growth Factor 9 (Fgf9). Recombinant Fgf9 protein inhibits the differentiation response of the mesenchyme to N-Shh, but does not affect proliferation. We propose a model for how factors made by two epithelial cell populations, the inner endoderm and the outer jacket of mesothelium, coordinately regulate the proliferation and differentiation of the lung mesoderm. PMID- 12781692 TI - Heparan sulfate-FGF10 interactions during lung morphogenesis. AB - Signaling by fibroblast growth factor 10 (FGF10) through FGFR2b is essential for lung development. Heparan sulfates (HS) are major modulators of growth factor binding and signaling present on cell surfaces and extracellular matrices of all tissues. Although recent studies provide evidence that HS are required for FGF directed tracheal morphogenesis in Drosophila, little is known about the HS role in FGF10-mediated bud formation in the vertebrate lung. Here, we mapped HS expression in the early lung and we investigated how HS interactions with FGF10 FGFR2b influence lung morphogenesis. Our data show that a specific set of HS low in O-sulfates is dynamically expressed in the lung mesenchyme at the sites of prospective budding near Fgf10-expressing areas. In turn, highly sulfated HS are present in basement membranes of branching epithelial tubules. We show that disrupting endogenous gradients of HS or altering HS sulfation in embryonic lung culture systems prevents FGF10 from inducing local responses and markedly alters lung pattern formation and gene expression. Experiments with selectively sulfated heparins indicate that O-sulfated groups in HS are critical for FGF10 signaling activation in the epithelium during lung bud formation, and that the effect of FGF10 in pattern is in part determined by regional distribution of O-sulfated HS. Moreover, we describe expression of a HS 6-O-sulfotransferase preferentially at the tips of branching tubules. Our data suggest that the ability of FGF10 to induce local budding is critically influenced by developmentally regulated regional patterns of HS sulfation. PMID- 12781693 TI - Continuing primordial germ cell differentiation in the mouse embryo is a cell intrinsic program sensitive to DNA methylation. AB - The initial cohort of mammalian gametes is established by the proliferation of primordial germ cells in the early embryo. Primordial germ cells first appear in extraembyronic tissues and subsequently migrate to the developing gonad. Soon after they arrive in the gonad, the germ cells cease dividing and undertake sexually dimorphic patterns of development. Male germ cells arrest mitotically, while female germ cells directly enter meiotic prophase I. These sex-specific differentiation events are imposed upon a group of sex-common differentiation events that are shared by XX and XY germ cells. We have studied the appearance of GCNA1, a postmigratory sex-common germ cell marker, in cultures of premigratory germ cells to investigate how this differentiation program is regulated. Cultures in which proliferation was either inhibited or stimulated displayed a similar extent of differentiation as controls, suggesting that some differentiation events are the result of a cell-intrinsic program and are independent of cell proliferation. We also found that GCNA1 expression was accelerated by agents which promote DNA demethylation or histone acetylation. These results suggest that genomic demethylation of proliferative phase primordial germ cells is a mechanism by which germ cell maturation is coordinated. PMID- 12781694 TI - Identification and characterization of stem cells in prepubertal spermatogenesis in mice. AB - The stem cell properties of gonocytes and prospermatogonia at prepubertal stages are still largely unknown: it is not clear whether gonocytes and prospermatogonia are a special cell type or similar to adult undifferentiated spermatogonia. To characterize these cells, we have established transgenic mice carrying EGFP (enhanced green fluorescence protein) cDNA under control of an Oct4 18-kb genomic fragment containing the minimal promoter and proximal and distal enhancers; Oct4 is reported to be expressed in undifferentiated spermatogonia at prepubertal stages. Generation of transgenic mice enabled us to purify gonocytes and prospermatogonia from the somatic cells of the testis. Transplantation studies of testicular cells so far have been done with a mixture of germ cells and somatic cells. This is the first report that establishes how to purify germ cells from total testicular cells, enabling evaluation of cell-autonomous repopulating activity of a subpopulation of prospermatogonia. We show that prospermatogonia differ markedly from adult spermatogonia in both the size of the KIT-negative population and cell cycle characteristics. The GFP(+) KIT(-) fraction of prospermatogonia has much higher repopulating activity than does the GFP(+)KIT(+) population in the adult environment. Interestingly, the GFP(+)KIT(+) population still exhibits repopulating activity, unlike adult KIT-positive spermatogonia. We also show that ALCAM, activated leukocyte cell adhesion molecule, is expressed transiently in gonocytes. Sertoli cells and myoid cells also express ALCAM at the same stage, suggesting that ALCAM may contribute to gonocyte-Sertoli cell adhesion and migration of gonoyctes toward the basement membrane. PMID- 12781695 TI - A gain-of-function mutation in oma-1, a C. elegans gene required for oocyte maturation, results in delayed degradation of maternal proteins and embryonic lethality. AB - In vertebrates, oocytes undergo maturation, arrest in metaphase II, and can then be fertilized by sperm. Fertilization initiates molecular events that lead to the activation of early embryonic development. In Caenorhabditis elegans, where no delay between oocyte maturation and fertilization is apparent, oocyte maturation and fertilization must be tightly coordinated. It is not clear what coordinates the transition from an oocyte to an embryo in C. elegans, but regulated turnover of oocyte-specific proteins contributes to the process. We describe here a gain of-function mutation (zu405) in a gene that is essential for oocyte maturation, oma-1. In wild type animals, OMA-1 protein is expressed at a high level exclusively in oocytes and newly fertilized embryos and is degraded rapidly after the first mitotic division. The zu405 mutation results in improper degradation of the OMA-1 protein in embryos. In oma-1(zu405) embryos, the C blastomere is transformed to the EMS blastomere fate, resulting in embryonic lethality. We show that degradation of several maternally supplied cell fate determinants, including SKN-1, PIE-1, MEX-3, and MEX-5, is delayed in oma-1(zu405) mutant embryos. In wild type embryos, SKN-1 functions in EMS for EMS blastomere fate specification. A decreased level of maternal SKN-1 protein in the C blastomere relative to EMS is believed to be responsible for this cell expressing the C, instead of the EMS, fate. Delayed degradation of maternal SKN-1 protein in oma-1(zu405) embryos and resultant elevated levels in C blastomere is likely responsible for the observed C-to-EMS blastomere fate transformation. These observations suggest that oma-1, in addition to its role in oocyte maturation, contributes to early embryonic development by regulating the temporal degradation of maternal proteins in early C. elegans embryos. PMID- 12781696 TI - The immunogenic peptide for Th1 development. AB - Th1 cells play a critical role in the induction of cell-mediated immune responses and eradication of intracellular pathogen. The dose and route of immunization of antigen are also determining factors. It remains unclear what types of immunogenic peptide can induce the Th1 development and how it acts to regulate the immune system. Ag85B (also known as alpha antigen or MPT59) has been shown to be the most potent antigen species yet purified in humans and in mice. Strong Th1 responses have been elicited in vitro from PPD(+) asymptomatic individuals and Ag85B-primed cells of C57BL/6 (I-A(b)) mice. Peptide-25 (aa240-254) of Ag85B is a major Th1 cell epitope in I-A(b) mice. Active immunization of C57BL/6 mice with Peptide-25 can induce the development of CDT4(+) TCRVbeta11(+) and CDT4(+) TCRVbeta11(-)Th1 cells that produce IFN-gamma- and TNF-alpha, and protects against subsequent infection with live Mycobacterium tuberculosis H37Rv IFN gamma. Peptide-25 has a potent adjuvant activity in both humoral and cell mediated immune responses that is mediated by Th1 cells. We would propose to designate Peptide-25 as "Th1-inducing peptide". PMID- 12781697 TI - Delivery of IL-12 intranasally leads to reduced IL-12-mediated toxicity. AB - Interleukin-12 (IL-12) is a heterodimeric cytokine that enhances immune responses to bacterial, parasitic, and viral pathogens, and leads to tumor regression in animal models. For this reason, the use of IL-12 as a vaccine adjuvant and as a therapeutic agent for the treatment of cancer is being investigated. Unfortunately, the extreme toxicity of this molecule observed during clinical trials has limited its use. This toxicity correlates with increased IFN-gamma expression, decreased glucose levels, and altered histological responses in the spleen and duodenum. In this study, we show that intranasal (i.n.) delivery of IL 12 is a less toxic route of inoculation compared to the commonly employed subcutaneous route. When delivered i.n., IL-12 induces less systemic IFN-gamma production and fewer pathological tissue changes, yet is efficacious, as indicated by enhanced CD3(+) T cell activation and increased production of Th1 associated immunoglobulins (i.e., serum IgG2a). Thus, IL-12 can be delivered safely and effectively by the i.n. route, a finding which may allow IL-12 to fulfill its clinical potential. PMID- 12781698 TI - Mechanism of activation of human peripheral blood NK cells at the single cell level by Echinacea water soluble extracts: recruitment of lymphocyte-target conjugates and killer cells and activation of programming for lysis. AB - Echinacea purpurea, a plant originally used by native Americans to treat respiratory infections, has also been shown to exert immunomodulatory activities both in vivo and in vitro. However, the mechanism underlying Echinacea-induced immunomodulation remains largely unknown. This study examined in vitro the effects of soluble extracts of E. purpurea on natural killer (NK) cells present in human peripheral blood mononuclear cells (PBMC). Flow cytometric methods were used to examine activation, cytotoxicity, NK-target binding, and killer cell frequency. Treatment of PBMC with Echinacea overnight resulted in the activation of CD69 expression and increase in mean fluorescence intensity in both the CD16+ and CD16+CD56+ NK subsets. However, the frequency of CD16+ cells was decreased as well as the mean fluorescence intensity was down-regulated. NK cytotoxicity was augmented 100% at the concentration of 0.1 microg/ml of Echinacea in a short time (4-h) assay. Examination at the single cell level revealed augmentation of the frequency of CD56+ NK-target conjugates and a plateau was reached after 30-60 min of incubation. Likewise, the frequency of CD56+ killer cells in the conjugates was also significantly increased by Echinacea. There was recruitment of non conjugated CD56+ cells into CD16+ NK-target conjugates and activation of the NK target non-killer conjugates into killer cells. These findings demonstrate that Echinacea extracts are potent activators of NK cytotoxicity. Echinacea augments the frequency of NK target conjugates and activates the programming for lysis of NK cells. PMID- 12781699 TI - Defense mechanisms of IFN-gamma and LPS-primed murine microglia against Acanthamoeba castellanii infection. AB - In the central nervous system (CNS), cytokine-primed microglia play a central role in host's defense against Acanthamoeba castellanii infection. In this study, the effect of recombinant interferon (rIFN)-gamma and Salmonella enterica serovar enteritidis lipopolysaccharide (LPS), both inflammatory stimuli, on A. castellanii infection in murine microglia was examined. Priming of microglia with rIFN-gamma and LPS synergistically triggered, in a dose-dependent manner, amebastatic activity in these cells. More than 52%, 88% or 95% of this function was then abrogated by anti-IL-1beta (but not anti-IL-1alpha), IL-6 or TNF-alpha neutralizing antibodies, suggesting that these endogenously produced cytokines may participate in the antimicrobial capacity. Consistent with these findings, the priming of microglia with rIFN-gamma and LPS elicited the release of proinflammatory interleukin (IL)-1alpha, IL-1beta, IL-6 and tumor necrosis factor (TNF)-alpha. Since L-canavanine affected amebastatic activity only during the priming process but not during the infection process, NO-dependent pathway appears to be not the sole antiparasitic mechanism involved in this function. These data suggest that rIFN-gamma and LPS, likely through a proinflammatory network, up-regulate the release of IL-beta, IL-6 and TNF-alpha, which could trigger antimicrobial activity against A. castellanii infection in the brain. PMID- 12781700 TI - Modulation of constitutive and delayed apoptosis by brefeldin A in human neutrophils. AB - Neutrophil apoptosis is a constitutive process that can be enhanced or delayed by various stimuli. In this study, the effect of brefeldin A (BFA), which affects the biological process of secretion, on constitutive and delayed apoptosis of neutrophils was investigated. Neutrophil apoptosis was determined after culturing for 20 h in vitro by morphological changes, annexin V staining, and DNA electrophoresis. BFA dose-dependently increased the constitutive apoptotic rate of neutrophils. The delay of apoptosis induced by granulocyte-macrophage colony stimulating factor (GM-CSF) and lipopolysaccharide (LPS) was also blocked by BFA. However, this effect of BFA was less marked when neutrophils were treated with dexamethasone, interleukin-8 (IL-8), or dibutyryl cAMP (dbcAMP). Moreover, the delay of neutrophil apoptosis induced by rottlerin, a specific inhibitor of protein kinase C (PKC)-delta, was significantly abrogated by BFA. Although BFA induced apoptosis was not blocked by the caspase-3 inhibitor, zDEVD-fmk, myeloid cell leukemia-1 (Mcl-1) expression levels were downregulated by BFA. These results suggest that derangement of vesicular protein transport may be involved in the apoptosis of neutrophils, and that the action of BFA on apoptosis is dependent on changes in the expression of Mcl-1. PMID- 12781701 TI - Inhibitory effects of glucocorticoids on rat eosinophil superoxide generation and chemotaxis. AB - Eosinophil infiltration into inflammatory tissues and the subsequent release of inflammatory mediators are the hallmarks of several inflammatory allergic diseases. Although there have been a considerable number of publications on anti inflammatory effects of glucocorticoids, little is known about whether glucocorticoids affect the activation of eosinophils directly. We studied the effects of three glucocorticoids, mometasone furoate, dexamethasone and beclomethasone dipropionate, on superoxide generation and the chemotaxis of rat eosinophils. Highly purified rat eosinophils were treated for 6 h with mometasone furoate, dexamethasone or beclomethasone dipropionate. Eosinophils were stimulated with phorbol myristate acetate (PMA) for superoxide generation, while for induction of chemotaxis, platelet-activating factor (PAF) or leukotriene B(4) (LTB(4)) was used. None of the glucocorticoids used in the present study caused significant suppressive effects on superoxide generation induced by PMA. On the other hand, both PAF- and LTB(4)-induced migration of rat eosinophils were inhibited in a concentration-dependent manner by glucocorticoids. Mometasone furoate showed a significant effect at concentrations higher than 10(-11) M. Dexamethasone and beclomethasone dipropionate also caused a significant inhibition at concentrations higher than 10(-8) and 10(-7) M, respectively. These results indicated that the anti-inflammatory effects of glucocorticoids were mediated by direct inhibition of eosinophil migration. Furthermore, mometasone furoate was suggested to be more useful than the other drugs in the treatment of allergic diseases responsible for eosinophil chemotaxis. PMID- 12781702 TI - Bartonella quintana lipopolysaccharide effects on leukocytes, CXC chemokines and apoptosis: a study on the human whole blood and a rat model. AB - Bartonella quintana, an emerging gram-negative pathogen, may cause trench fever, endocarditis, cerebral abscess and bacillary angiomatosis usually with the absence of septic shock in humans. B. quintana lipopolysaccharide (LPS), a deep rough endotoxin with strong reactivity in the limulus amebocyte lysate (LAL) assay, was studied in human whole blood and in a rat model. A significant (P<0.05) increase of interleukin-8 (IL-8) concentration, comparable to the level induced by enterobacterial LPS, was stimulated in the human whole blood by B. quintana LPS. Isolated human neutrophils delayed their apoptotic behavior in the presence of B. quintana LPS. In the rat, B. quintana LPS induced a significant (P<0.001) increase in white blood cell count, both 30 and 60 min after intravenous injection. Such leukocytosis was inhibited by pretreatment with prazosin, an alpha-adrenergic antagonist. B. quintana LPS did not significantly change heart rate (HR), hematocrit (HCT) and platelet count in the above reported in vivo model, and regarding mean blood pressure (MAP) only a very early (5 min after LPS) and mild (yet significant) hypotension was observed. In contrast, a long-lasting decrease of MAP was found in Salmonella minnesota R595 LPS-treated animals. Blood TNFalpha levels did not change significantly from the baseline in rats injected with either saline or with B. quintana LPS, on the contrary S. minnesota R595 LPS-injected animals showed substantial increase of TNFalpha levels up to 2924 pg/ml at 60 min after LPS injection. B. quintana LPS as well as Salmonella LPS-injected rats exhibited an increase of the blood levels of GRO/CINC-1, particularly at 240 min after LPS administration. Apical part of rat gut villi showed several TUNEL-positive cells in tissue sections from B. quintana LPS-treated animals. Taken together, our data demonstrates that B. quintana LPS is able to selectively stimulate some inflammatory mediators. B. quintana LPS induced leukocytosis appears mediated by an alpha-adrenergic receptor. The delayed apoptotic process of leukocytes and the chemokine increase may explain the apoptotic cells found in the rat gut and the inflammatory reactions in some human Bartonella diseases. This peculiar inflammatory pattern induced by B. quintana LPS, may partially account for the lack of severe septic shock, observed in human B. quintana infections. PMID- 12781703 TI - Exogenous but not endogenous prostanoids regulate cytokine secretion from murine bone marrow dendritic cells: EP2, DP, and IP but not EP1, EP3, and FP prostanoid receptors are involved. AB - Murine bone marrow-derived dendritic cells (DC), stimulated with lipopolysaccharide (LPS) and/or LPS+interferon-gamma (IFN-gamma), secrete a variety of inflammatory mediators which may modulate their functions. We have examined the potential for exogenous prostanoids, acting in a paracrine fashion, and endogenous prostanoids, acting in an autocrine fashion, to regulate secretion of tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), IL-10, and IL 12 in DC. In order to identify receptors mediating these effects, DC were treated in vitro with receptor-selective prostanoids. Agonists of cyclic AMP-elevating receptors, namely, prostaglandin E(2) (PGE(2)), butaprost (EP(2) receptor), iloprost (IP receptor), and BW245C (DP receptor), dose-dependently inhibited the release of IL-6, TNF-alpha, and IL-12 and enhanced the release of IL-10 from LPS stimulated DC, with TNF-alpha secretion being the most strongly affected. In contrast, 15-deoxy-Delta(12,14)-PGJ(2)-an activator of nuclear peroxisome proliferator-activated receptor-gamma (PPAR-gamma) receptors-inhibited release of all tested cytokines. Exogenous prostanoids, cyclic AMP-elevating analogs, lost their ability to modulate cytokine release in cells pre-incubated for 4 h with LPS, indicating that prostanoids may affect DC functions during initial phases of LPS stimulation only. Sulprostone and (+)-fluprostenol failed to modulate any of responses tested, suggesting lack of involvement/expression of EP(1), EP(3), and FP receptors in DC activation. In order to examine the role of endogenous prostanoids, DC were treated with inhibitors of cyclooxygenases (COX). At concentrations that completely blocked PGE(2) release, neither indomethacin (nonselective inhibitor) nor rofecoxib (COX-2-selective inhibitor) influenced cytokine release from LPS-stimulated DC. Thus, cytokine release from LPS stimulated DC does not seem to be autoregulated by endogenous prostanoids, whereas in vivo regulatory function may be fulfilled in a paracrine manner by PGD(2), PGE(2), and PGI(2) released from neighboring cells. PMID- 12781704 TI - Mechanism of inhibition of T-acute lymphoblastic leukemia cells by PNP inhibitor- BCX-1777. AB - Purine nucleoside phosphorylase (PNP) deficiency in humans produces a relatively selective depletion of T cells. BCX-1777 is a potent inhibitor of PNP. BCX-1777 in the presence of deoxyguanosine (dGuo) inhibits the proliferation of CEM-SS [T acute lymphoblastic leukemia (T-ALL)] cells with an IC(50)=0.015 microM. This inhibition by BCX-1777 and dGuo is accompanied by elevation of dGTP (154-fold) and dATP (8-fold). Deoxycytidine (dCyt) completely and lamivudine (3TC) partially reverse this inhibition caused by BCX-1777 and dGuo. dNTP analysis of these samples indicates that, in the presence of dCyt, where complete reversal of inhibition is observed, dGTP and dATP pools revert back to the control levels. In samples containing 3TC, where partial reversal of inhibition was observed, dGTP decreased from 154-fold to 38-fold and dATP levels further increased from 8-fold to 30-fold compared to the control sample. In CEM-SS cells, inhibition of proliferation by BCX-1777 and dGuo is not due to accumulation of dATP because in the presence of 3TC, where reversal of inhibition is observed, dATP levels are further increased. These studies clearly indicate that inhibition of T cells is due to accumulation of dGTP resulting in cell death with characteristics of apoptosis. The half-life of dGTP in CEM-SS cells is 18 h, which is longer than that observed in human lymphocytes (4 h), suggesting that the nucleotidase level in CEM-SS cells is lower than in human lymphocytes. A 154-fold accumulation of dGTP in CEM-SS cells in the presence of BCX-1777 and dGuo compared to a 15-fold accumulation of dGTP in human lymphocytes suggests that kinase level is higher in CEM-SS cells compared to human lymphocytes. High kinase and low nucleotidase levels make CEM-SS cells more sensitive to inhibition by BCX-1777 and dGuo than human lymphocytes. Currently, BCX-1777 is in phase I/II clinical trial for the treatment of T cell malignancies. PMID- 12781705 TI - Protective effects of Chlorella vulgaris in lead-exposed mice infected with Listeria monocytogenes. AB - Chlorella vulgaris extract (CVE) was examined for its chelating effects on the myelosuppression induced by lead in Listeria monocytogenes-infected mice. The reduction in the number of bone marrow granulocyte-macrophage progenitors (CFU GM) observed after the infection was more severe in the groups previously exposed to lead. Extramedullar hematopoiesis, which was drastically increased after the infection, was not altered by the presence of lead. Treatment with CVE, given simultaneously or following lead exposure, restored to control values the myelosuppression observed in infected/lead-exposed mice and produced a significant increase in serum colony-stimulating activity. The benefits of the CVE treatment were also evident in the recovery of thymus weight, since the reduction produced by the infection was further potentiated by lead exposure. The efficacy of CVE was evident when infected and infected/lead-exposed mice were challenged with a lethal dose of L. monocytogenes after a 10-day treatment with 50 mg/kg CVE/day, given simultaneously to the exposure to 1300 ppm lead acetate in drinking water. Survival rates of 30% for the infected group and of 20% for the infected/lead-exposed groups were observed. Evidence that these protective effects of CVE are partly due to its chelating effect was given by the changes observed in blood lead levels. We have observed in the group receiving the CVE/lead simultaneous exposure a dramatic reduction of 66.03% in blood lead levels, when compared to lead-exposed nontreated control. On the other hand, CVE treatment following lead exposure produced a much less effective chelating effect. CVE treatments for 3 or 10 days, starting 24 h following lead exposure, produced a reduction in blood lead levels of 13.5% and 17%, respectively, compared to lead-exposed nontreated controls. The significantly better response observed with the simultaneous CVE/lead administration indicates that the immunomodulation effect of CVE plays an important role in the ability of this algae to reduce blood lead levels. In this regard, additional experiments with gene knockout C57BL/6 mice lacking a functional IFN-gamma gene demonstrated that this cytokine is of paramount importance in the protection afforded by CVE. The antibacterial evaluation measured by the rate of survival demonstrated that, in face of a 100% survival in the control group composed of normal C57BL/6 mice, which are resistant to L. monocytogenes, we observed no protection whatsoever in the IFN-gamma knockout C57BL/6 mice treated with CVE and inoculated with L. monocytogenes. PMID- 12781706 TI - Bestatin selectively suppresses the growth of leukemic stem/progenitor cells with BCR/ABL mRNA transcript in patients with chronic myelogeneous leukemia. AB - The in vitro effect of bestatin on Philadelphia (Ph) chromosome positive chronic myeloid leukemia (CML) was investigated using mature clonogenic cells and primitive stem cells derived from long-term culture-initiating cells (LTC-ICs). Individual colonies were grown in methycellulose culture (clonogenic cells) and after 5 weeks, LTC (colonies derived from LTC-ICs) were individually isolated. DNA isolated from these clonogenic colonies was studied by reverse transcriptase polymerase chain reaction (RT-PCR) analysis to detect BCR/ABL mRNA transcripts b3a2 and b2a2. At the mature hematopoietic progenitor cell level, almost all (20/21) colonies, including both erythroid and myeloid progenitors, were leukemic, i.e. BCR/ABL mRNA positive. Although normal progenitors were able to grow in the presence of bestatin, even at the most primitive progenitor cell level (LTC-ICs), the number of leukemic clones gradually decreased. Furthermore, bestatin suppressed the outgrowth of leukemic clones more frequently than control LTC without any effect on the growth of normal clones. These results indicate that bestatin, at levels that can be obtained by per os administration clinically, suppresses only Ph-positive leukemic clones without affecting normal hematopoiesis. Based on these results, we suggest that bestatin has the potential to provide another treatment for patients with CML. PMID- 12781707 TI - Synergistic autoactivation of the Epstein-Barr virus immediate-early BRLF1 promoter by Rta and Zta. AB - Expression of two Epstein-Barr virus (EBV) immediate-early gene products, Zta (encoded by the BZLF1 gene) and Rta (encoded by the BRLF1 gene), are required for the switch from latent infection to virus replication. We have analyzed the regions of the BRLF1 gene promoter (Rp) that are required for Rta and Zta transactivation of Rp. Notably, significant synergy between the actions of Rta and Zta on Rp was observed in both a B cell line (DG75) and an epithelial cell line (293), suggesting that during induction of the viral lytic cycle low levels of these viral transactivators are likely sufficient to initiate the entire lytic cascade. However, while two Zta binding sites (ZREs) have been identified in Rp, the proximal ZRE was the dominant site for mediating Zta transactivation. Rta activation of Rp was diminished by mutation of the proximal Sp1 binding site, as previously reported (J. Virol. 75 (2001), 5240), but mutation of this site only had a modest impact on transactivation of Rp by Rta in the presence of Zta. Further deletion analyses of Rp failed to identify a critical site for Rta transactivation of Rp in the presence of Zta, with the exception of deleting the TATAA box of Rp, suggesting that a non-DNA binding mechanism may be involved in the observed activation of Rp by Rta. We also observed promiscuous activation of several reporter constructs by Rta, suggesting that Rta activation of gene expression may involve a general non-DNA binding mechanism. Decreasing the amount of transfected Rta expression vector reduced background Rta activation, while retaining specific activation of Rp. PMID- 12781708 TI - A longitudinal assessment of autologous neutralizing antibodies in children perinatally infected with human immunodeficiency virus type 1. AB - The evolution of autologous neutralizing antibodies to sequential human immunodeficiency virus type 1 (HIV-1) isolates was studied in a population of 16 children who were perinatally infected with human immunodeficiency virus type 1. The cohort included seven children with rapid disease progression (RP) and nine who had nonrapid disease progression (NRP). Four of the NRP after 6 months of age harbored viruses that could be neutralized by antibodies found in autologous contemporaneous plasma (titers up to 1:640) while the majority of longitudinally collected viruses from five NRP were resistant to neutralization with contemporaneous plasma. Because of their shorter survival, only five of the RP had studies after 6 months of age; three of the five had neutralizing antibodies to contemporaneous virus isolates and the highest titers were 1:20. The highest titers in RP (up to 1:160) occurred in specimens obtained prior to 6 months of age but these were most likely of maternal origin. Most isolates that were not neutralized by contemporaneous plasma could be neutralized using noncontemporaneous plasma obtained months to years after the virus isolates. These autologous noncontemporaneous neutralizing antibodies persisted for years, had titers that were higher to viruses isolated at younger ages, and were generally more potent in children with NRP than RP. Demonstration of neutralizing antibodies to viruses previously resistant to neutralization by contemporaneous plasma suggests a continuous evolution of virus variants in vivo that are able to escape the effect of neutralizing antibodies. PMID- 12781709 TI - Detection of a group 2 coronavirus in dogs with canine infectious respiratory disease. AB - An investigation into the causes of canine infectious respiratory disease was carried out in a large rehoming kennel. Tissue samples taken from the respiratory tract of diseased dogs were tested for the presence of coronaviruses using RT-PCR with conserved primers for the polymerase gene. Sequence analysis of four positive samples showed the presence of a coronavirus with high similarity to both bovine and human coronavirus (strain OC43) in their polymerase and spike genes, whereas there was a low similarity to comparable genes in the enteric canine coronavirus. This canine respiratory coronavirus (CRCV) was detected by RT PCR in 32/119 tracheal and 20/119 lung samples, with the highest prevalence being detected in dogs with mild clinical symptoms. Serological analysis showed that the presence of antibodies against CRCV on the day of entry into the kennel decreased the risk of developing respiratory disease. PMID- 12781710 TI - Deletion of pe38 attenuates AcMNPV genome replication, budded virus production, and virulence in heliothis virescens. AB - The pe38 gene product of Autographa californica M nucleopolyhedrovirus (AcMNPV) has been shown to be involved in transcriptionally transactivating viral genes and augmenting viral DNA replication in transient assays. To assess the role of pe38 during infection, we generated a knockout virus, Delta pe38-E9/E9, in which the pe38 open reading frame was replaced with that of the green fluorescent protein. We compared mutant and wild-type (WT) viral replication in insect cell culture and virulence in Heliothis virescens larvae. Compared to WT, Delta pe38 E9/E9 budded virus (BV) production was delayed by at least 3 h, and BV yields were reduced over 99%. Similarly, Delta pe38-E9/E9 DNA synthesis levels were greatly reduced relative to those of WT, but onset of DNA replication was the same for both viruses. In bioassays, nearly sevenfold more Delta pe38-E9/E9 virus than WT virus was required to achieve an LD(50) when administered orally, but not hemocoelically. These results support the hypothesis that the kinetics of AcMNPV BV production greatly impact virulence in larvae infected orally (the natural route of infection) and that PE38 is an important, but not essential, factor in viral DNA synthesis and BV production. PMID- 12781711 TI - Features of the Env leader protein and the N-terminal Gag domain of feline foamy virus important for virus morphogenesis. AB - Previous studies have shown that foamy virus (FV) particle budding, especially the involvement of the viral env glycoprotein is different from that of other (ortho) retroviruses: the N-terminal Env leader protein Elp is a constituent of released FV particles. A defined sequence in Elp required for particle budding binds to the MA domain of Gag. To extend these findings, we show that feline FV Elp is a membrane-anchored protein with the N-terminus located inside the particle. Thus, the internal/cytoplasmic domain of Elp has the correct topology for interacting with Gag during budding. In addition to Elp, an Elp-related protein of about 9 kDa was shown to be virion associated and is probably generated by cellular signal peptidases. Besides the function of Elp binding, the N-terminal domain of Gag was shown to be required for proper localization of feline FV Gag to the cytoplasm and the perinuclear/nuclear region. PMID- 12781712 TI - Borna disease virus induces acute fatal neurological disorders in neonatal gerbils without virus- and immune-mediated cell destructions. AB - Borna disease virus (BDV) is a noncytolytic, neurotropic RNA virus that is known to cause neurological disturbances in various animal species. Our previous experiment demonstrated that neonate gerbils develop an acute fatal neurological disease following infection with BDV, Virology 282, 65-76). The study suggested that BDV directly causes functional damage of neuronal cells resulting in the lethal disorder in neonatal gerbils. To extend this finding, we examined whether BDV can induce neurological diseases in the absence of virus- and immune-mediated cell destruction, by using cyclosporine A (CsA)-treated neonatal gerbils. Although CsA completely suppressed specific antibody production and brain inflammation in the infected gerbil brains, the fatal neurological disorder was not inhibited by the treatment. Furthermore, we demonstrated that CsA treatment significantly decreased brain levels of cytokines, except interleukin (IL)-1 beta, in the infected gerbils. These results suggested that BDV replication, as well as brain cytokines, at least IL-1 beta, rapidly induces fatal disturbances in gerbil brain. We demonstrate here that BDV exhibits a unique neuropathogenesis in neonatal gerbil that may be pathologically and immunologically different from those in two other established rodent models, rats and mice. With this novel rodent model of virus infection it should be possible not only to examine acute neurological disturbances without severe neuroanatomical and immunopathological alterations but also to analyze molecular and cellular damage by virus replication in the central nervous system. PMID- 12781713 TI - Biological and genetic characteristics of HIV infections in Cameroon reveals dual group M and O infections and a correlation between SI-inducing phenotype of the predominant CRF02_AG variant and disease stage. AB - In Yaounde, Cameroon, HIV-1 group-specific V3 serology on 1469 HIV-positive samples collected between 1996 and 2001 revealed that group O infections remained constant around 1% for 6 years. Only one group N sample was identified and 4.3% reacted with group M and O peptides. Although the sensitivity of the group specific polymerase chain reaction (PCR) in two genomic regions was not optimal, we confirmed, in at least 6 of 49 (12.2%) dual O/M seropositive samples and in 1 of 9 group O samples, dual infection with group O and M viruses (n = 4) or with group O or M virus and an intergroup recombinant virus (n = 3). Partial env (V3 V5) sequences on a subset of 295 samples showed that at least eight subtypes and five circulating recombinant forms (CRFs) of HIV-1 group M co-circulate; more than 60% were CRF02_AG and 11% had discordant subtype/CRF designations between env and gag. Similarly as for subtype B, the proportion of syncytium-inducing strains increased when CD4 counts were low in CRF02_AG-infected patients. The V3 loop charge was significantly lower for non-syncytium-inducing strains than for syncytium-inducing strains but cannot be used as an individual marker to predict phenotype. The two predominant HIV-1 variants in Africa, CRF02_AG and subtype C, thus have different biological characteristics. PMID- 12781714 TI - Probing the ability of the coat and vertex protein of the membrane-containing bacteriophage PRD1 to display a meningococcal epitope. AB - Bacteriophage PRD1 is an icosahedral dsDNA virus with a diameter of 740 A and an outer protein shell composed of 720 copies of major coat protein P3. Spike complexes at the vertices are composed of a pentameric base (protein P31) and a spike structure (proteins P5 and P2) where the N-terminal region of the trimeric P5 is associated with the base and the C-terminal region of P5 is associated with receptor-binding protein P2. The functionality of proteins P3 and P5 was investigated using insertions and deletions. It was observed that P3 did not tolerate changes whereas P5 tolerated changes much more freely. These properties support the hypothesis that viruses have core structures and functions, which remain stable over time, as well as other elements, responsible for host interactions, which are evolutionally more fluid. The insertional probe used was the apex of exposed loop 4 of group B meningococcal outer membrane protein PorA, a medically important subunit vaccine candidate. It was demonstrated that the epitope could be displayed on the virus surface as part of spike protein P5. PMID- 12781715 TI - A functional interferon regulatory factor-1 (IRF-1)-binding site in the upstream regulatory region (URR) of human papillomavirus type 16. AB - We recently identified a new enhancer element (HIRE-1, HPV-Interferon Responsive Element-1) in the upstream regulatory region (URR) of human papillomavirus (HPV) type 16. HIRE-1 is located upstream from and in close proximity to the TATA box. HIRE-1 is 1 nt shorter in its 5' sequence in comparison to a consensus IRF-1 binding site (IRF-E). Gel shift analyses clearly demonstrated that HIRE-1 is capable of binding IRF-1 in response to interferon-gamma (IFN-gamma) treatment. In a reporter system, HIRE-1 stimulated transcription in response to IRF-1 or IFN gamma from both a heterologous or the homologous (p97) promoter in a dose dependent manner. Mutations in the core binding sequence strongly decreased this enhancer activity. Interestingly, HIRE-1 stimulated transcription in the context of the full URR in a cell-type-specific manner, thereby suggesting the role of other cell-type-specific factors that might counteract with its function. Thus, our results may explain the inconsistent clinical and experimental results observed following IFN treatment of cervical lesions or cells. Also, this new enhancer may have an important function during inflammatory responses against HPV type 16. PMID- 12781716 TI - The three-dimensional structure of cocksfoot mottle virus at 2.7 A resolution. AB - Cocksfoot mottle virus is a plant virus that belongs to the genus Sobemovirus. The structure of the virus has been determined at 2.7 A resolution. The icosahedral capsid has T = 3 quasisymmetry and 180 copies of the coat protein. Except for a couple of stacked bases, the viral RNA is not visible in the electron density map. The coat protein has a jelly-roll beta-sandwich fold and its conformation is very similar to that of other sobemoviruses and tobacco necrosis virus. The N-terminal arm of one of the three quasiequivalent subunits is partly ordered and follows the same path in the capsid as the arm in rice yellow mottle virus, another sobemovirus. In other sobemoviruses, the ordered arm follows a different path, but in both cases the arms from three subunits meet and form a similar structure at a threefold axis. A comparison of the structures and sequences of viruses in this family shows that the only conserved parts of the protein-protein interfaces are those that form binding sites for calcium ions. Still, the relative orientations and position of the subunits are maintained. PMID- 12781718 TI - Specific in vitro cleavage of Mason-Pfizer monkey virus capsid protein: evidence for a potential role of retroviral protease in early stages of infection. AB - Processing of Gag polyproteins by viral protease (PR) leads to reorganization of immature retroviral particles and formation of a ribonucleoprotein core. In some retroviruses, such as HIV and RSV, cleavage of a spacer peptide separating capsid and nucleocapsid proteins is essential for the core formation. We show here that no similar spacer peptide is present in the capsid-nucleocapsid (CA-NC) region of Mason-Pfizer monkey virus (M-PMV) and that the CA protein is cleaved in vitro by the PR within the major homology region (MHR) and the NC protein in several sites at the N-terminus. The CA cleavage product was also identified shortly after penetration of M-PMV into COS cells, suggesting that the protease-catalyzed cleavage is involved in core disintegration. PMID- 12781717 TI - Defective RNAs of Citrus tristeza virus analogous to Crinivirus genomic RNAs. AB - The family Closteroviridae includes the genera Closterovirus and Ampelovirus with monopartite genomes and the genus Crinivirus with bipartite genomes. Plants infected with the Closterovirus, Citrus tristeza virus (CTV), often contain one or more populations of defective RNAs (dRNAs). Although most dRNAs are comparatively small (2-5 kb) consisting of the genomic RNA termini with large internal deletions, we recently characterized large dRNAs of approximately 12 kb that retained the open reading frames (ORFs) 1a plus 1b. These were self replicating RNAs and appeared to be analogous to the genomic RNA 1 of the bipartite criniviruses. The present report describes the finding of an additional group of large dRNAs (LdRNAs) that retained all or most of the 10 3' ORFs and appeared to be analogous to genomic RNA 2 of criniviruses. Isolates associated with LdRNAs were found associated with double-recombinant dRNAs (DR-dRNAs) of various sizes (1.7 to 5.1 kb) that comprised the two termini and a noncontiguous internal sequence from ORF2. The genetic and epidemiological implications of the architectural identities of LdRNAs and DR dRNAs and their apparent analogy with the genomic RNA 2 of criniviruses are discussed. PMID- 12781719 TI - Membrane fusion activity of vesicular stomatitis virus glycoprotein G is induced by low pH but not by heat or denaturant. AB - The fusogenic envelope glycoprotein G of the rhabdovirus vesicular stomatitis virus (VSV) induces membrane fusion at acidic pH. At acidic pH the G protein undergoes a major structural reorganization leading to the fusogenic conformation. However, unlike other viral fusion proteins, the low-pH-induced conformational change of VSV G is completely reversible. As well, the presence of an alpha-helical coiled-coil motif required for fusion by a number of viral and cellular fusion proteins was not predicted in VSV G protein by using a number of algorithms. Results of pH dependence of the thermal stability of G protein as determined by intrinsic Trp fluorescence and circular dichroism (CD) spectroscopy show that the G protein is equally stable at neutral or acidic pH. Destabilization of G structure at neutral pH with either heat or urea did not induce membrane fusion or conformational change(s) leading to membrane fusion. Taken together, these data suggest that the mechanism of VSV G-induced fusion is distinct from the fusion mechanism of fusion proteins that involve a coiled-coil motif. PMID- 12781720 TI - The effect of ribavirin and IMPDH inhibitors on hepatitis C virus subgenomic replicon RNA. AB - The recent development of in vitro hepatitis C virus (HCV) RNA replication systems has provided useful tools for studying the intracellular anti-HCV activity of ribavirin. Ribavirin has been shown to: (1) induce "error catastrophe" in poliovirus, Proc. Natl. Acad. Sci. USA 98, 6895-6900), (2) be a pseudo-substrate of the HCV RNA-dependent RNA polymerase (RdRp) in vitro, J. Biol. Chem. 276, 46094-46098), and (3) increase mutations in HCV RNA in the binary T7 polymerase/HCV cDNA replication system, J. Virol. 76, 8505-8517). These findings have led to the hypothesis that ribavirin may also induce error catastrophe in HCV. However, the functional relevance of ribavirin-induced HCV RNA mutagenesis is unclear. By use of a colony formation assay, in which RNA is isolated from the HCV subgenomic replicon system following treatment, the impact of ribavirin, inosine-5'-monophosphate dehydrogenase (IMPDH) inhibitors, and the combination was assessed. Ribavirin reduced HCV replicon colony-forming efficiency (CFE) in a dose-dependent fashion, suggesting that ribavirin may be misincorporated into replicon RNA and result in an anti-replicon effect analogous to error catastrophe. This effect was markedly suppressed by addition of exogenous guanosine. Combination treatment with ribavirin and mycophenolic acid (MPA) or VX-497, both potent, nonnucleoside IMPDH inhibitors, led to a greatly enhanced anti-replicon effect. This enhancement was reversed by inclusion of guanosine with the treatment. In contrast, MPA or VX-497 alone had only marginal effects on both the quantity and quality (CFE) of replicon RNA, suggesting that although IMPDH inhibition is an important contributing factor to the overall ribavirin anti-HCV replicon activity, IMPDH inhibition by itself is not sufficient to exert an anti-HCV effect. Sequencing data targeting the neo gene segment of the HCV replicon indicated that ribavirin together with MPA or VX-497 increased the replicon error rate by about two-fold. Taken together these results further suggest that lethal mutagenesis may be an effective anti-HCV strategy. The colony formation assay provides a useful tool for evaluating mutagenic nucleoside analogs for HCV therapy. Finally, the data from combination treatment indicate potential therapeutic value for an enhanced anti-HCV effect when using ribavirin in combination with IMPDH inhibition. PMID- 12781721 TI - A novel method for analysis of membrane microdomains: vesicular stomatitis virus glycoprotein microdomains change in size during infection, and those outside of budding sites resemble sites of virus budding. AB - Membrane proteins, including viral envelope glycoproteins, may be organized into areas of locally high concentration, commonly referred to as membrane microdomains. Some viruses bud from detergent-resistant microdomains referred to as lipid rafts. However, vesicular stomatitis virus (VSV) serves as a prototype for viruses that bud from areas of plasma membrane that are not detergent resistant. We developed a new analytical method for immunoelectron microscopy data to determine whether the VSV envelope glycoprotein (G protein) is organized into plasma membrane microdomains. This method was used to quantify the distribution of the G protein in microdomains in areas of plasma membrane that did not contain budding sites. These microdomains were compared to budding virus envelopes to address the question of whether G protein-containing microdomains were formed only at the sites of budding. At early times postinfection, most of the G protein was organized into membrane microdomains outside of virus budding sites that were approximately 100-150 nm, with smaller amounts distributed into larger microdomains. In contrast to early times postinfection, the increased level of G protein in the host plasma membrane at later times postinfection led to distribution of G protein among membrane microdomains of a wider variety of sizes, rather than a higher G protein concentration in the 100- to 150-nm microdomains. VSV budding occurred in G protein-containing microdomains with a range of sizes, some of which were smaller than the virus envelope. These microdomains extended in size to a maximum of 300-400 nm from the tip of the budding virion. The data support a model for virus assembly in which G protein organizes into membrane microdomains that resemble virus envelopes prior to formation of budding sites, and these microdomains serve as the sites of assembly of internal virion components. PMID- 12781722 TI - The complete sequence of marine bacteriophage VpV262 infecting vibrio parahaemolyticus indicates that an ancestral component of a T7 viral supergroup is widespread in the marine environment. AB - The 46,012-bp sequence of the marine bacteriophage VpV262 infecting the bacterium Vibrio parahaemolyticus is reported. The VpV262 sequence reveals that it is a distant relative of marine Roseophage SIO1, and an even more distant relative of coliphage T7. VpV262 and SIO1 appear to represent a widespread marine phage group that lacks an RNA polymerase gene and is ancestral to the T7-like phages. We propose that this group together with the T7-like phages be designated as the T7 supergroup. The ancestral head structure gene module for the T7 supergroup was reconstructed by using sensitive biased Psi-blast searches supplemented by statistical support derived from gene order. In the early and replicative segments, these phages have participated in extensive interchange with the viral gene pool. VpV262 carries a different replicative module than SIO1 and the T7 like phages. PMID- 12781723 TI - Multimodal integration of high-resolution EEG and functional magnetic resonance imaging data: a simulation study. AB - Previous simulation studies have stressed the importance of the use of fMRI priors in the estimation of cortical current density. However, no systematic variations of signal-to-noise ratio (SNR) and number of electrodes were explicitly taken into account in the estimation process. In this simulation study we considered the utility of including information as estimated from fMRI. This was done by using as the dependent variable both the correlation coefficient and the relative error between the imposed and the estimated waveforms at the level of cortical region of interests (ROI). A realistic head and cortical surface model was used. Factors used in the simulations were the different values of SNR of the scalp-generated data, the different inverse operators used to estimated the cortical source activity, the strengths of the fMRI priors in the fMRI-based inverse operators, and the number of scalp electrodes used in the analysis. Analysis of variance results suggested that all the considered factors significantly afflict the correlation and the relative error between the estimated and the simulated cortical activity. For the ROIs analyzed with simulated fMRI hot spots, it was observed that the best estimation of cortical source currents was performed with the inverse operators that used fMRI information. When the ROIs analyzed do not present fMRI hot spots, both standard (i.e., minimum norm) and fMRI-based inverse operators returned statistically equivalent correlation and relative error values. PMID- 12781724 TI - Comparison of functional activation foci in children and adults using a common stereotactic space. AB - The development of methods allowing direct comparisons between child and adult neuroimaging data is an important prerequisite for studying the neural bases of cognitive development. Several issues arise when attempting to make such direct comparisons, including the comparability of anatomical localization of functional responses and the magnitude and time course of the hemodynamic responses themselves. Previous results suggest that, after transformation into a common stereotactic space, anatomical differences between children (ages 7 and 8) and adults are small relative to the resolution of fMRI data. Here, we investigate whether time courses (BOLD responses) and locations of functional activation foci show similarities as well. Event-related fMRI was performed on 16 children (ages 7 and 8) and 16 adults, who pressed buttons in response to a visual stimulus. After transforming images into Talairach space, the coordinates of four consistent activations in each hemisphere were determined for each subject: two foci in the sensorimotor cortex, one focus in the visual cortex, and one focus in the supplementary motor area (eight activations in total). In seven foci, time courses were similar between children and adults, and peak amplitudes of time courses were comparable in all eight foci. There were negligible between-group differences in location of all foci. Variability of activation location was statistically similar in the two groups. In voxelwise group comparison images, minimal differences were found between children and adults in visual and motor cortex regions. The small differences in time courses and locations of activation foci between child and adult brains validate the feasibility of direct statistical comparison of these groups within a common space. PMID- 12781725 TI - Sustained and transient activity during an object-naming task: a mixed blocked and event-related fMRI study. AB - Cognitive tasks often involve at least two types of processes-sustained processes potentially related to ongoing task demands and transient processes related to the processing of individual items within the task. Using functional magnetic resonance imaging, in conjunction with a mixed-blocked and event-related design, we examined sustained and transient patterns of neural activity during an object naming task. Subjects were imaged during runs that alternated between control blocks and task blocks. During task blocks, primed and unprimed objects were intermixed and jittered in time. Regions of interest based on separate analyses of sustained and transient activities were tested independently for sustained and transient responses. Three general patterns of results were observed. (1) Some regions exhibited transient responses but little or no sustained response. These regions were widely distributed across the brain. (2) Other regions clearly exhibited both transient and sustained responses. These regions were found primarily in lateral and medial frontal lobes. (3) A few regions exhibited a sustained response but little or no transient responses. These regions were found in the basal ganglia, orbitofrontal lobe, and right lateral frontal lobe. Furthermore, two homotopic regional pairs in the right and left inferior frontal lobe (frontal operculum and inferior frontal cortex) showed a crossover of sustained and transient effects, with greater transient activity in the left and greater sustained activity in the right hemisphere. The asymmetric relationship between sustained and transient responses in prefrontal regions may be an example of task-specific biasing at work. PMID- 12781726 TI - Detection of cortical transition regions utilizing statistical analyses of excess masses. AB - A new statistical approach for observer-assisted detection of transition regions of adjacent cytoarchitectonic areas within the human cerebral cortex was developed. This method analyzes the structural information of cytoarchitectural profiles (e.g., the modality of a gray level intensity distribution) based on observed excess mass differences verified by a suitable statistical test. Profiles were generated by scanning the cerebral cortex over respective regions of interest that were oriented to trajectories running parallel to the orientation of cell columns. For each single profile, determination of excess masses provided evidence for a certain number of peaks in the cell density, thereby avoiding fluctuation due solely to sampling anomalies. Comparing such excess mass measurements by means of multiple local rank tests over a wide range of profiles allowed for the detection of cytoarchitectural inhomogeneities at respective given confidence levels. Special parameters (e.g., level of significance, width of targeted region, number of peaks) then could be adapted to specific pattern recognition problems in lamination analyses. Such analyses of excess masses provided a general tool for observer-assisted evaluation of profile arrays. This observer-assisted statistical method was applied to five different cortical examples. It detected the same transition regions that had been determined earlier through direct examination of samples, despite cortical convexities, concavities, and some minor staining inhomogeneities. PMID- 12781727 TI - Overlapping neural regions for processing rapid temporal cues in speech and nonspeech signals. AB - Speech perception involves recovering the phonetic form of speech from a dynamic auditory signal containing both time-varying and steady-state cues. We examined the roles of inferior frontal and superior temporal cortex in processing these aspects of auditory speech and nonspeech signals. Event-related functional magnetic resonance imaging was used to record activation in superior temporal gyrus (STG) and inferior frontal gyrus (IFG) while participants discriminated pairs of either speech syllables or nonspeech tones. Speech stimuli differed in either the consonant or the vowel portion of the syllable, whereas the nonspeech signals consisted of sinewave tones differing along either a dynamic or a spectral dimension. Analyses failed to identify regions of activation that clearly contrasted the speech and nonspeech conditions. However, we did identify regions in the posterior portion of left and right STG and left IFG yielding greater activation for both speech and nonspeech conditions that involved rapid temporal discrimination, compared to speech and nonspeech conditions involving spectral discrimination. The results suggest that, when semantic and lexical factors are adequately ruled out, there is significant overlap in the brain regions involved in processing the rapid temporal characteristics of both speech and nonspeech signals. PMID- 12781728 TI - Amygdalar atrophy in panic disorder patients detected by volumetric magnetic resonance imaging. AB - It has been suggested that the pathophysiology of panic disorder (PD) may involve abnormalities in several brain structures, including the amygdala. To date, however, no study has used quantitative structural neuroimaging techniques to examine amygdalar anatomy in this disorder. Volumetric magnetic resonance imaging (MRI) studies of the amygdalas, hippocampi, and temporal lobes were conducted in 12 drug-free, symptomatic PD patients (six females and six males), and 12 case matched healthy comparison subjects. Volumetric MRI data were normalized for brain size. PD patients were found to have smaller left-sided and right-sided amygdalar volumes than controls. No differences were found in either hippocampi or temporal lobes. These findings provide new evidence of changes in amygdalar structure in PD and warrant further anatomical and MRI brain studies of patients with this disorder. PMID- 12781729 TI - Spatiotemporal maps of past-tense verb inflection. AB - Does the brain inflect verbs by applying rules, by associative retrieval of the inflected form, or both? We used whole-head magnetoencephalography to spatiotemporally map the brain response underlying verb past-tense inflection. Placing either regular or irregular verbs into the past tense sequentially modulates the bilateral visual, left inferotemporal, posterior superior temporal (Wernicke's area), left inferior prefrontal (Broca's area), and right prefrontal cortices. Although irregular and regular verb inflection evokes similar cortical response patterns, differences in specific frontotemporal regions are observed. At approximately 340 ms, irregular verbs evoke greater response modulation in left occipitotemporal cortex. This modulation occurs when widespread areas are simultaneously active, suggesting that it reflects associative activation necessary for generation of past-tense forms. Subsequently, regular verbs show increased response at approximately 470 ms within left inferior prefrontal regions associated with rule-based inflection. Increased right dorsolateral prefrontal response at approximately 570 ms may represent directed/effortful retrieval of irregular past-tense forms. Thus, the brain inflects verbs by dynamically modulating different functional divisions of an integrated language system. PMID- 12781730 TI - Individual differences in rCBF correlates of syntactic processing in sentence comprehension: effects of working memory and speed of processing. AB - Positron emission tomography (PET) was used to determine the effect of working memory and speed of sentence processing on regional cerebral blood flow (rCBF) during syntactic processing in sentence comprehension. PET activity associated with making plausibility judgments about syntactically more complex subject object (SO) sentences (e.g., The juice that the child spilled stained the rug) was compared to that associated with making judgments about synonymous syntactically simpler object-subject (OS) sentences (e.g., The child spilled the juice that stained the rug). Two groups of nine subjects differing in working memory and matched for speed of sentence processing both showed increases in rCBF in lateral posteroinferior frontal lobe bilaterally. The subjects were reclassified to form two groups of eight subjects who were matched for working memory but who differed in speed of sentence processing. Fast-performing subjects activated lateral posteroinferior frontal lobe bilaterally and slow-performing subjects showed activation of left superior temporal lobe. The results indicate that rCBF responses to syntactic comprehension tasks vary as a function of speed of sentence processing but not as a function of working memory. PMID- 12781731 TI - Learning-induced neural plasticity associated with improved identification performance after training of a difficult second-language phonetic contrast. AB - Adult native Japanese speakers have difficulty perceiving the English /r-l/ phonetic contrast even after years of exposure. However, after extensive perceptual identification training, long-lasting improvement in identification performance can be attained. This fMRI study investigates localized changes in brain activity associated with 1 month of extensive feedback-based perceptual identification training by native Japanese speakers learning the English /r-l/ phonetic contrast. Before and after training, separate functional brain imaging sessions were conducted for identification of the English /r-l/ contrast (difficult for Japanese speakers), /b-g/ contrast (easy), and /b-v/ contrast (difficult), in which signal-correlated noise served as the reference control condition. Neural plasticity, denoted by exclusive enhancement in brain activity for the /r-l/ contrast, does not involve only reorganization in brain regions concerned with acoustic-phonetic processing (superior and medial temporal areas) but also the recruitment of additional bilateral cortical (supramarginal gyrus, planum temporale, Broca's area, premotor cortex, supplementary motor area) and subcortical regions (cerebellum, basal ganglia, substantia nigra) involved with auditory-articulatory (perceptual-motor) mappings related to verbal speech processing and learning. Contrary to what one may expect, brain activity for perception of a difficult contrast does not come to resemble that of an easy contrast as learning proceeds. Rather, the results support the hypothesis that improved identification performance may be due to the acquisition of auditory articulatory mappings allowing for perception to be made in reference to potential action. PMID- 12781732 TI - Increased regional cerebral blood flow but normal distribution of GABAA receptor in the visual cortex of subjects with early-onset blindness. AB - Before the completion of visual development, visual deprivation impairs synaptic elimination in the visual cortex. The purpose of this study was to determine whether the distribution of central benzodiazepine receptor (BZR) is also altered in the visual cortex in subjects with early-onset blindness. Positron emission tomography was carried out with [(15)O]water and [(11)C]flumazenil on six blind subjects and seven sighted controls at rest. We found that the CBF was significantly higher in the visual cortex for the early-onset blind subjects than for the sighted control subjects. However, there was no significant difference in the BZR distribution in the visual cortex for the subject with early-onset blindness than for the sighted control subjects. These results demonstrated that early visual deprivation does not affect the distribution of GABA(A) receptors in the visual cortex with the sensitivity of our measurements. Synaptic elimination may be independent of visual experience in the GABAergic system of the human visual cortex during visual development. PMID- 12781733 TI - The spatial extent of the BOLD response. AB - Functional magnetic resonance imaging is routinely used to localize brain function, with multiple brain scans averaged together to reveal activation volumes. In this study, we examine the seldom-studied effect of multiple scan averaging on the extent of activation volume. Using restricted visual field stimulation, we obtained a large number of scan repetitions and analyzed changes in activation volume with progressively increased averaging and across single scans. Activation volume increased monotonically with averaging and failed to asymptote when as many as 22 scans were averaged together. Expansions in the spatial extent of activation were not random; rather, they were centered about activation loci that appear with little or no averaging. Using empirical and simulated data, changes with averaging in activation volumes and cross correlation coefficient distributions revealed the presence of considerably more activated voxels than commonly surmised. Many voxels have low SNR and remain undetected without extensive averaging. The primary source of such voxels was not downstream venous drainage since there was no significant and consistent delay difference between voxels activated at different averaging levels. Voxels with low SNR may reflect a diffuse subthreshold activity centered about spiking neurons, dephasing gradients from distal veins, or simply a blood flow response extending beyond the locus of neuronal firing. Across single scans, as much as twofold changes in activation volume were observed. These changes were not correlated with the order of scan acquisition, subject task performance, or signal and noise properties of activated voxels. Instead, they may reflect subtle changes between overlapping noise and signal frequency components. PMID- 12781734 TI - Dopamine receptors and transporters in the brain reward circuits of type 1 and 2 alcoholics measured with human whole hemisphere autoradiography. AB - The role of the dopamine system in brain reward mechanisms and development of substance abuse is well-established with nucleus accumbens as a key structure in mediating these effects. Several studies on alcoholism have indicated defects in dopaminergic neurotransmission and alterations in dopamine receptor densities. However, it has remained unclear if the substance abuse-related dopaminergic defect is specifically associated with a certain receptor subtype. The aim of this study was to compare putative alterations of dopamine D(1,) D(2), and D(3) receptors in nucleus accumbens, amygdala, and substantia nigra among alcoholics and controls. We studied the densities of dopamine D(1) and D(3) receptors in brains of 9 type 1 alcoholics, 8 type 2 alcoholics, and 10 healthy controls by using postmortem human whole hemisphere autoradiography. The mean densities of dopamine D(1) and D(3) receptors were at the same level in all study groups. Combining these with our previous results, our data suggest that among type 1 alcoholics dopamine transporters are lower in nucleus accumbens and dopamine D(2), but not D(1) or D(3) receptors in nucleus accumbens and amygdala. Further, the densities of all these dopamine-binding sites among type 2 alcoholics are at the level of healthy controls. The results suggest that lower dopamine receptor density is specific for D(2) receptor and for type 1 alcoholism, which supports Cloninger's neurogenetic model of two alcoholic subtypes, and indicates the importance of classifying these subgroups separately when issues related to dopaminergic activity are studied. PMID- 12781735 TI - An event-related optical topography study of cortical activation induced by single-pulse transcranial magnetic stimulation. AB - To visualize cortical activations during transcranial magnetic stimulation (TMS), it is necessary to measure those activations at high spatiotemporal resolution while preventing interference with the magnetic property of a coil. One suitable method that satisfies these demands is optical topography (OT), which has been used in cortical activation studies. In the present study, single-pulse TMS was applied to the left primary motor area, and cortical responses at the stimulation site were measured simultaneously with event-related OT. When TMS was applied at 110% motor threshold (MT), we observed significant oxyhemoglobin increases that were both time-locked and correlated with the hemodynamic basis function. Moreover, when TMS was applied at 90% MT, significant oxyhemoglobin increases were detected even though there were no motor-evoked potentials. These results demonstrate that OT can directly measure cortical responses to subthreshold single-pulse TMS, independent of the afferent feedback from the peripheral neuromuscular activity. PMID- 12781736 TI - The functional anatomy of parkinsonian bradykinesia. AB - To investigate the difficulty that patients with Parkinson's disease (PD) have in performing fast movements, we used H(2)(15)O PET to study regional cerebral blood flow (rCBF) associated with performance of a simple predictive visuomanual tracking task at three different velocities. Tracking movements in PD patients (versus tracking with the eyes alone) were associated with a general underactivation of the areas normally activated by the task (sensorimotor cortex contralateral to the moving arm, bilateral dorsal premotor cortices, and ipsilateral cerebellum). Presupplementary motor cortex (pre-SMA) ipsilateral to the moving arm had greater than normal movement-related activations. Increasing movement velocity led to increased rCBF in multiple premotor and parietal cortical areas and basal ganglia in the patients as opposed to the few cerebral locations that are normally velocity-related. The functional correlates of PD bradykinesia are: (1) impaired recruitment of cortical and subcortical systems that normally regulate kinematic parameters of movement such as velocity; and (2) increased recruitment of multiple premotor areas including both regions specialized for visuomotor control (ventral premotor and parietal cortices) and some that are not (pre-SMA). The overactivation of cortical regions observed in patients may be functional correlates of compensatory mechanisms and/or impaired suppression as a facet of the primary pathophysiology of PD. PMID- 12781737 TI - Anatomic dissociation of selective and suppressive processes in visual attention. AB - Visual spatial attention is associated with activation in parietal regions as well as with modulation of visual activity in ventral occipital cortex. Within the parietal lobe, localisation of activity has been hampered by variation in individual anatomy. Using fMRI within regions of interest derived from individual functional maps, we examined the response of superior parietal lobule, intraparietal sulcus, and ventral occipital cortex in 11 normal adults as attention was directed to the left and right visual hemifields during bilateral visual stimulation. Activation in ventral occipital cortex was augmented contralateral to the attended hemifield (P < 0.006), while intraparietal activation was augmented ipsilaterally (P < 0.009), and superior parietal lobule showed no modulation of activity as a function of attended hemifield. These findings suggest that spatial enhancement of relevant stimuli in ventral occipital cortex is complemented by an intraparietal response associated with suppression of, or preparation of a reflexive shift of attention toward, irrelevant stimuli. The spatial attention system in superior parietal cortex, in contrast, may be driven to equal degrees by currently attended stimuli and by stimuli that are potential targets of attention. PMID- 12781738 TI - Method for functional MRI mapping of nonlinear response. AB - Nonlinear systems analysis combining blood oxygen level dependent (BOLD), functional magnetic resonance imaging (fMRI) and m-sequence stimulation paradigms are proposed as a new method for exploring neuronal responses and interactions. Previous studies of electrical activity in the human visual cortex have observed significant nonlinearities of task-induced activity with temporal dynamics on a timescale of 10-20 ms. Despite the confounding effect of the seconds-long hemodynamic response, it is demonstrated that BOLD fMRI can be used to probe neuronal interactions on a time scale of tens of ms. Visual activation experiments were performed with various stimuli, and amplitude maps of first and second order kernel coefficients were generated using correlation analysis. Second order nonlinearities in BOLD fMRI were observed and attributed to temporal contrast caused by transitions in the stimulus sequence. In addition, the kernel maps showed significant differences between second order nonlinearities of foveal and peripheral vision. By including a reference experiment with a slightly modified stimulus presentation, a distinction could be made between (fast) neuronal nonlinearities and hemodynamic effects on the time scale of the seconds. The results indicate that BOLD fMRI can probe fast neuronal nonlinearities. PMID- 12781739 TI - Modeling regional and psychophysiologic interactions in fMRI: the importance of hemodynamic deconvolution. AB - The analysis of functional magnetic resonance imaging (fMRI) time-series data can provide information not only about task-related activity, but also about the connectivity (functional or effective) among regions and the influences of behavioral or physiologic states on that connectivity. Similar analyses have been performed in other imaging modalities, such as positron emission tomography. However, fMRI is unique because the information about the underlying neuronal activity is filtered or convolved with a hemodynamic response function. Previous studies of regional connectivity in fMRI have overlooked this convolution and have assumed that the observed hemodynamic response approximates the neuronal response. In this article, this assumption is revisited using estimates of underlying neuronal activity. These estimates use a parametric empirical Bayes formulation for hemodynamic deconvolution. PMID- 12781740 TI - Dynamic responses of protein homeostatic regulatory mechanisms to perturbations from steady state. AB - Nineteen hypothetical protein homeostatic regulatory mechanisms were constructed and analysed in terms of the rate at which they recovered from a perturbation in the steady-state concentration of any component. Systems were constructed to symbolize transcription/translation processes of the average protein from Escherichia coli (1000 copies of protein P along with 1 gene G per cell). In some model systems, G catalysed the synthesis of P directly, while in others G catalysed the synthesis of mRNA (called M), and M catalysed the synthesis of P in a subsequent step. Recovery rates for each regulatory mechanism were obtained by generating the corresponding system of differential equations, linearizing the system about the steady state, and determining eigenvalues of the associated coefficient matrix. The optimal rate of recovery for a given mechanism, R(D), was determined by combining random and gradient search approaches to find rate constants for which the system recovered fastest. Regulatory elements that improved dynamic regulation were identified. These consisted of negative feedback relationships that involved P binding to either G (to shut off the synthesis of P) or M (to stimulate its degradation). Regulation improved as increasing numbers of P's bound to either G or M; however, the binding to M was more effective. In other mechanisms PP dimers bound G. Dimer-binding mechanisms were roughly twice as effective in terms of regulation as those that bound P monomers. The effect of linking two regulatory "modules" was also investigated. Linking had no effect on R(D), but optimal rate constants for the linked system were similar to those of the unlinked modules, suggesting that it may be feasible to construct regulatory networks by linking individual modules of this type. PMID- 12781741 TI - Founder control and coexistence in a simple model of asymmetric competition for light. AB - Size asymmetry in plant light acquisition complicates predictions of competitive outcomes in light-limited communities. We present a mathematically tractable model of asymmetric competition for light and discuss its implications for predicting outcomes of competition during establishment in two-, three-, and many species communities. In contrast to the resource-reduction model of symmetric competition for a single resource, the model we present predicts that outcomes of asymmetric competition for light will sometimes depend on the timing of establishment and the consequent hierarchy among species in canopy position. Competitive outcomes in the model depend on the minimum light requirements (L(c)) and self-shading of species lower in the canopy compared to the light available (L(out)(*)) beneath species higher in the canopy. Succession progresses towards species with decreasing values for L(c), but arrested successions occur when initial dominants have relatively high values for L(c) but low values for L(out)(*), leading to founder control. A theoretically limitless number of species may coexist in competition for light when dominance is founder controlled. These model predictions have implications for an array of applied ecological questions, including methods to control invasive species in light limited restored ecosystems. PMID- 12781742 TI - Traveling waves in a model of influenza A drift. AB - Between major pandemics, the influenza A virus changes its antigenic properties by accumulating point mutations (drift) mainly in the RNA genes that code for the surface proteins hemagglutinin (HA) and neuraminidase (NA). The successful strain (variant) that will cause the next epidemic is selected from a reduced number of progenies that possess relatively high transmissibility and the ability to escape from the immune surveillance of the host. In this paper, we analyse a one dimensional model of influenza A drift (Z. Angew. Math. Mech. 76 (2) (1996) 421) that generalizes the classical SIR model by including mutation as a diffusion process in a phenotype space of variants. The model exhibits traveling wave solutions with an asymptotic wave speed that matches well those obtained from numerical simulations. As exact solutions for these waves are not available, asymptotic estimates for the amplitudes of infected and recovered classes are provided through an exponential approximation based on the smallness of the diffusion constant. Through this approximation, we find simple scaling properties to several parameters of relevance to the epidemiology of the disease. PMID- 12781743 TI - Advanced formulation of base pair changes in the stem regions of ribosomal RNAs; its application to mitochondrial rRNAs for resolving the phylogeny of animals. AB - The ribosomal RNAs (rRNAs) of animal mitochondria, especially those of arthropod mitochondria, have a higher content of G:U and U:G base pairs in their stem regions than the nuclear rRNAs. Thus, the theoretical formulation of base pair changes is extended to incorporate the faster base pair changes A:U<-->G:U<-->G:C and U:A<-->U:G<-->C:G into the previous formulation of the slower base pair changes between A:U, G:C, C:G and U:A. The relative base pair change probability containing the faster and slower base pair changes is theoretically derived to estimate the divergence time of rRNAs under the influence of selection for these base pairs. Using the cartilaginous fish-teleost fish divergence and the crustacean-insect divergence as calibration points, the present method successfully predicts the divergence times of the main branches of animals: Deuterostomia and Protostomia diverged 9.2 x 10(8) years ago, the divergence of Echinodermata, Hemichordata and Cephalochordata succeedingly occurred during the period from 8 x 10(8) to 6 x 10(8) years ago, while Arthropoda, Annelida and Mollusca diverged almost concomitantly about 7 x 10(8) years ago. The dating for the divergence of Platyhelminthes and Cnidaria is traced back to 1.2 x 10(9) years ago. This result is consistent with the fossil records in the Stirling Range Formation of southwestern Australia, the Ediacara and Avalon faunas and the Cambrian Burgess Shale. Thus, the present method may be useful for estimating the divergence times of animals ranging from 10(8) to 10(9) years ago, resolving the difficult problems, e.g. deviation from rate constancy and large sampling variances, in the usual methods of treating apparent change rates between individual bases and/or base pairs. PMID- 12781744 TI - Can transitive inference evolve in animals playing the hawk-dove game? AB - What should an individual do if there are no reliable cues to the strength of a competitor when fighting with it for resources? We herein examine the evolutionarily stable strategy (ESS) in the hawk-dove game, if the opponent's resource-holding potential (RHP) can only indirectly be inferred from the outcome of past interactions in the population. The strategies we examined include the classical mixed strategy in which no information on past games is utilized, the 'imprinting' strategy in which a player increases/decreases its aggressiveness if it wins/loses a game, the 'immediate inference' strategy in which a player can infer the strength of those opponents it fought before, and the 'transitive inference' strategy in which a player can infer the strength of a new opponent through a third party with which both players have fought before. Invasibility analysis for each pair of strategies revealed that (i) the transitive-inference strategy can always invade the mixed strategy and the imprinting strategy, and itself refuses invasion by these strategies; (ii) the largest advantage for transitive inference is achieved when the number of games played per individual in one generation is small and when the cost of losing an escalated game is large; (iii) the immediate inference, rather than the transitive inference, can be an ESS if the cost of fighting is small; (iv) a strong linear ranking is established in the population of transitive-inference strategists, though it does not perfectly correlate to the ranking by actual RHPs. We found that the advantage of the transitive inference is not in its ability to correct a misassessment (it is actually the worst in doing so), but in the ability of quickly lining up either incorrect or correct assessments to form a linear dominance hierarchy. PMID- 12781745 TI - The gene-orientation structure of eukaryotes. AB - It is shown that the sequence of gene orientations of four eukaryotes-for those that are presently known-are well modeled by a two-state, two-parameter (first order) Markov chain. These include the six chromosomes of nematodes (C. elegans), the 16 chromosomes of yeast (S. cerevisiae), the five chromosomes of Arabidopsis (A. thaliana), and the 19 scaffolds of fruit flies (D. melanogaster). Moreover, they are reasonably well modeled, more simply, by a one-parameter symmetric version of the Markov chain. Further, compelling statistical evidence is presented which suggests that the parameters particularizing the Markov chain are organism dependent rather than merely chromosome dependent. This surprising observation begs an appropriate biological explanation. Does there exist some kind of mechanism of "communication" among a eukaryote's chromosomes that serves to maintain common values for all of the chromosomal parameters? Or are the common parameter values merely a consequence of a common environment of origin for all of the chromosomes of an organism? (If so, why so?) A third possible explanation is ruled out: while there exists within the class of Markov chain models under consideration, a case that could be described as equivalent to "flipping a fair coin," a thorough-random-shuffling-of-the-genes explanation, via mutations, does not hold up to statistical scrutiny. PMID- 12781746 TI - Selective pressures on genomes in molecular evolution. AB - We describe the evolution of macromolecules as an information transmission process and apply tools from Shannon information theory to it. This allows us to isolate three independent, competing selective pressures that we term compression, transmission, and neutrality selection. The first two affect genome length: the pressure to conserve resources by compressing the code, and the pressure to acquire additional information that improves the channel, increasing the rate of information transmission into each offspring. Noisy transmission channels (replication with mutations) give rise to a third pressure that acts on the actual encoding of information; it maximizes the fraction of mutations that are neutral with respect to the phenotype. This neutrality selection has important implications for the evolution of evolvability. We demonstrate each selective pressure in experiments with digital organisms. PMID- 12781747 TI - A theoretical and empirical investigation of delayed growth response in the continuous culture of bacteria. AB - When the growth of bacteria in a chemostat is controlled by limiting the supply of a single essential nutrient, the growth rate is affected both by the concentration of this nutrient in the culture medium and by the amount of time that it takes for the chemical and physiological processes that result in the production of new biomass. Thus, although the uptake of nutrient by cells is an essentially instantaneous process, the addition of new biomass is delayed by the amount of time that it takes to metabolize the nutrient. Mathematical models that incorporate this "delayed growth response" (DGR) phenomenon have been developed and analysed. However, because they are formulated in terms of parameters that are difficult to measure directly, these models are of limited value to experimentalists. In this paper, we introduce a DGR model that is formulated in terms of measurable parameters. In addition, we provide for this model a complete set of criteria for determining persistence versus extinction of the bacterial culture in the chemostat. Specifically, we show that DGR plays a role in determining persistence versus extinction only under certain ranges of chemostat operating parameters. It is also shown, however, that DGR plays a role in determining the steady-state nutrient and bacteria concentrations in all instances of persistence. The steady state and transient behavior of solutions of our model is found to be in agreement with data that we obtained in growing Escherichia coli 23716 in a chemostat with glucose as a limiting nutrient. One of the theoretical predictions of our model that does not occur in other DGR models is that under certain conditions a large delay in growth response might actually have a positive effect on the bacteria's ability to persist. PMID- 12781748 TI - Gene arrangements and branching orders of gram-positive bacteria. AB - The availability of complete genomic sequence data allows one to develop new methods of reconstructing phylogenetic trees. A simple method of reconstructing branching orders based on gene transposition (or lateral transfer) is presented. It is argued that specific gene arrangements on four different genomes could determine a branching order. A computer search for such gene arrangements was carried out against gene order data of completely sequenced Gram-positive bacteria. Gene arrangements around ribosomal protein S4 gene, murC (UDP-N acetylmuramate:alanine ligase) gene and dnaE (DNA polymerase III alpha chain) gene each suggest a branching order in which actinobacteria with a high genomic G+C content first branched off from other Gram-positives with a low G+C content and then a split occurred between Mycoplasma species and a group closely related to Bacillus subtilis. A recently sequenced thermophilic bacterium Thermoanaerobacter tengcongensis is suggested to have branched off from the lineage leading to the low G+C Gram-positives prior to the split between the Mycoplasma and Bacillus groups. By contrast to the indel analysis in which a single evolutionary event of insertion or deletion of a signature sequence is assumed, the present method does not necessarily require such a parsimonious assumption of gene transposition. PMID- 12781749 TI - Factors affecting binary sex evolution with respect to avoidance of vertical transmission of deleterious intracellular parasites. AB - In sexual reproductive systems, the number of sexes is generally binary, viz. male and female. Several theoretical studies have shown that the evolution of this system is possibly related to cytoplasmic DNA, including deleterious cytoplasmic symbionts. When organisms are infected by a symbiont that is transmitted vertically to offspring via gametes, the exclusion or degeneration of the latter may evolve as a characteristic of those organisms. If this necessarily results in the elimination of organelle DNA in gametes, a reciprocal preference between individuals, one transmitting organelles and the other not, may be favored. In this theoretical study, factors affecting such an evolutionary process, in which the symbiont is considered as a parasite infecting vertically, horizontally and naturally, are considered. In addition, host individuals are assumed to recover from the infection to some degree. According to the analysis, a binary sex system can evolve only when uninfected and infected host individuals co-exist in a single host population. This condition can be satisfied only if natural infection occurs. Although recovery from infection has both positive and negative effects on binary sex evolution, the latter is promoted only when natural infection exists. Accordingly, if natural infection does not exist, the evolution of binary sex system is unlikely with respect to deleterious cytoplasmic symbionts, in absent of heterozogotic advantage in vertical transmission. PMID- 12781750 TI - Regime changes in ecological systems: an information theory approach. AB - We present our efforts at developing an ecological system index using information theory. Specifically, we derive an expression for Fisher Information based on sampling of the system trajectory as it evolves in the space defined by the state variables of the system, i.e. its state space. The Fisher Information index, as we have derived it, is a measure of system order, and captures the characteristic variation in speed and acceleration along the system's periodic steady-state trajectories. When calculated repeatedly over the system period, this index tracks steady states and transient behavior. We believe that such an index could be useful in detecting system 'flips' associated with a regime change, i.e. determining when systems are in a transient between one steady state and another. We illustrate the concepts using model ecosystems. PMID- 12781751 TI - Local perceptions of risk to livelihood in semi-arid Tanzania. AB - This paper examines variation in local perceptions of risk in semi-arid Tanzania, identifying factors that influence local perceptions of problems and testing the feasibility of risk mapping as a technique. Twelve villages in six districts were visited between February and April 2001. Villagers were asked about their worries and concerns in providing for themselves and their families using a risk questionnaire. The responses were grouped into 21 categories of problem and incidence and severity indices were calculated for each category. Most problems were associated with the availability of natural resources on which livelihoods depend but others related to human and social capital assets. In addition to environmental factors, livelihood strategy and gender both influenced people's perceptions of risk. Problems of irrigation and weather, for example, were important for agricultural communities while problems relating to livestock diseases, access to land and hunger were more important in pastoral communities. The risks cited by men and women generally reflected their traditional roles in society. Very broadly, the risks associated with natural capital tended to be seen as higher by men while women mentioned more problems relating to human and social capital. However, this was not always the case. Problems linked to finance, traditionally the concern of men, were ranked similarly by men and women. In projects designed to facilitate community management and control over common pool resources, the identification of common interests is particularly important. Risk mapping can provide a cost-effective way of gaining insights to help improve research design and to inform policy development. PMID- 12781752 TI - Ten common mistakes in designing biodiversity indicators for forest policy. AB - This paper identifies 10 common 'mistakes' in developing and using forest biodiversity indicators from the standpoint of making better forest management choices. The mistakes relate to a failure to clarify the values-basis for indicator selection and a failure to integrate science and values to design indicators that are concise, relevant and meaningful to decision makers. The combined effects of these ten mistakes include inconsistent and indefensible on ground management strategies and hidden trade-offs at a policy level. They result in frustrated professionals, a confused public, an inability to assess performance with respect to key forest policy objectives and, almost certainly, types and amounts of biodiversity conservation that fail to achieve either scientifically or socially preferred levels. Correcting the mistakes will help to address these problems and, more generally, recognizes the need to better understand the interface between science, public values, and decision making. PMID- 12781753 TI - Community modelling: a tool for correlating estimates of exposure with perception of odour from municipal solid waste (MSW) landfills. AB - Odours from municipal solid waste landfills have the potential to cause significant annoyance and impact to amenity in the environment surrounding sites. In order to assess the impact of odorous emissions on surrounding communities a quantitative model to predict annoyance was developed. The overall objective of this research was to develop the major components of the model namely, assessment of odorous emissions, dispersion and reception by the surrounding community around the landfill site. This study used community modelling as a tool to find a link between dispersion and perception of odour. The research completed a year round monitoring program engaging people within the local community as regular odour monitors. Estimates of exposure from dispersion analysis were used to compare incidents causing complaint and intensity-concentration plots were fitted for each monitor whose reports were found to be logically consistent with regard to the intensity scaling. Human responses to the vast range of odour intensities, from highly intense source odours through to less intense dispersed odours at monitors' locations, were found to differ greatly. It was observed that the psychophysical models based on the Weber-Fechner law and Power law fitted the data consistently well for the entire range of the intensity scale used, 1-7. However, the other two models, based on Beidler's law and Laffort's equation showed an inconsistency with the intensity scales >3. PMID- 12781754 TI - Economic transition and environmental sustainability: effects of economic restructuring on air pollution in the Russian Federation. AB - Economic liberalization in former socialist countries may have various implications for their environmental sustainability. Positive effects of this process are potentially associated with improved efficiency, investments into cleaner technologies, responsiveness to environmentally aware markets, and ending subsidies to heavy industries. On the other hand, market liberalization may result in weaker environmental controls, economic instabilities distracting attention from environmental issues, and increasing orientation towards profit making leading to more intensive exploitation of natural resources. In addition, trade liberalization may result in shifts towards more pollution and resource intensive industries. This article seeks to quantify effects of economic restructuring in Russia on air pollution from productive economic sectors in the 1990s. Air pollution in Russia had significantly declined in 1991-1999, however, this decline was largely due to economic decline, as the overall pollution intensity of the economy had decreased only slightly. The factors that affected the pollution intensity are: (1) a decrease in the combined share of industrial and transport activities in the economy and (2) changing pollution intensities of the industrial and transport sectors. The pollution intensity of the Russian industry had remained relatively stable during the 1990s. This was the result of the two opposite and mutually canceling trends: (a) increasing shares of pollution-intensive branches such as metal smelting and oil production vs. less pollution intensive manufacturing and (b) decline in pollution intensities within the industrial branches. The article proposes a methodology by which the contribution of both factors to the overall pollution intensity of the industrial sector can be quantified. The pollution intensity of the Russian transport sector appears to have declined in the first half of the 1990s and increased in the second half. The most recent trend can be explained by a rising proportion of private motorcars used for transportation of people and goods instead of traditional rail and other public transport. The findings of the paper demonstrate that shifts towards more pollution-, resource- and energy-intensive industries as a result of economic liberalization emerges as a significant negative factor of the process of economic transition threatening sustainability of emerging market economies. A research agenda to further investigate these impacts is proposed. PMID- 12781755 TI - Dispersion of odour: a case study with a municipal solid waste landfill site in North London, United Kingdom. AB - Municipal solid waste (MSW) landfills are a potential source of offensive odours that can create annoyance within communities. Dispersion modelling was used to quantify the potential odour strength causing an impact on the community around a particular MSW landfill site north of the London area in the United Kingdom. The case studies were completed with the short-term mode of COMPLEX-I, software developed by the US-EPA. The year 1998 was chosen as a source of baseline data. It was observed that by 2004, when the landfill will progress towards the west and a big band of the area towards the north would be partly/fully restored, the maximum contribution of the new sources giving higher odour concentrations would be in the southwesterly regions away from the landfill. Concentrations as high as 25.0 ou(E)/m(3) were observed with 3 min averaging time in the southwesterly areas as compared to concentrations of 20.0 ou(E)/m(3) at 10 min averaging times. However, the percentage frequency of such critical events occurring would be low. All other surrounding farms and small villages would be exposed to the concentration of 3.0 ou(E)/m(3) on certain occasions. In the year 2008, the majority of the filling fronts would be filled with wastes with no contributions from the active and operational cells. The maximum odour concentration around the landfill site for 1 h averaging time would be approximately 3 ou(E)/m(3) about 1.0 km north and 500 m west of the landfill site. For 3 min averaging time, the stretch of 5 ou(E)/m(3) band would be up to 2.5 km towards the north of the landfill site. It is argued that further analysis of the model calculations considering effects of wind direction, frequency of wind direction, stability of the atmosphere, selected odour threshold, integration time of the model, etc. would form a basis for calculating the separation distances of the landfill site from the surrounding community. PMID- 12781756 TI - Hedgerow planting analysed as a social system--interaction between farmers and other actors in Denmark. AB - In a number of European countries, including Denmark, the practice of planting hedgerows has a long tradition, and hedgerows form significant structures of semi natural elements in a matrix of arable land. In Denmark an institutionalised framework has developed in relation to a subsidy scheme encouraging farmers to plant hedgerows. This article analyses the planting practice as a social system; giving emphasis to the interactions between actors and how this affects the current planting activity. Combining an overall description of the nationwide network with a detailed case study in Jutland, Denmark enables an understanding of how the local planting practice is influenced by the local context and the nationwide network, and at the same time contributes to the reproduction of the entire network. It is concluded that the planting activity is characterised by routines, professionalism and division of labour. The local actors involved perceive the activity as a success, and do not question current practise. However, the actors are not aware of the potential consequences of the planting practice at the landscape scale. Even though hedgerows are planted through so called planting associations related to defined local areas the placement of hedgerows is not co-ordinated and evaluated at a landscape scale. In addition, the composition of hedgerows is standardised at a national level, and the individual farmer makes few adjustments. Thus, the sense of local landscape identity may become blurred. The increasing public attention towards a multiplicity of functions in the agricultural landscape implies further development of the potential positive landscape effects of hedgerows in a local context. This may induce changes in the relationships between actors and eventually the planting practice. PMID- 12781757 TI - Public perceptions and attitudes towards an established managed realignment scheme: Orplands, Essex, UK. AB - Where appropriate, managed realignment is the preferred policy option for sustainable coastal defences in the UK, since it provides both economic and ecological advantages. Until recently however, research has failed to address the social implications of this policy option having only acknowledged that communities in general, show reluctance in conceding land to the sea. Orplands managed realignment scheme is the third in a series of case studies to address a variety of social issues, but with a particular focus on the public acceptance of schemes and public confidence in the Environment Agency. It is hypothesised that local residents will be more accepting of a realignment scheme the longer it is in the public domain, rather than at its inception or construction. This paper presents findings from the Orplands scheme and demonstrates that overall scheme acceptance was greater at this site in comparison to previous case studies. Despite this, respondents did not show any more confidence in the Environment Agency and this has been attributed to their general scepticism towards a statutory body and the lack of recent contact between the various stakeholders. It is suggested that the development of criteria for consultation and public participation would facilitate public acceptance of managed realignment schemes. PMID- 12781758 TI - Environmental impacts of modern agricultural technology diffusion in Bangladesh: an analysis of farmers' perceptions and their determinants. AB - Farmers' perception of the environmental impacts of modern agricultural technology diffusion and factors determining such awareness were examined using survey data from 21 villages in three agro-ecological regions of Bangladesh. Results reveal that farmers are well aware of the adverse environmental impacts of modern agricultural technology, although their awareness remains confined within visible impacts such as soil fertility, fish catches, and health effects. Their perception of intangible impacts such as, toxicity in water and soils is weak. Level and duration of modern agricultural technology adoption directly influence awareness of its adverse effects. Education and extension contacts also play an important role in raising awareness. Awareness is higher among farmers in developed regions, fertile locations and those with access to off-farm income sources. Promotion of education and strengthening extension services will boost farmers' environmental awareness. Infrastructure development and measures to replenish depleting soil fertility will also play a positive role in raising awareness. PMID- 12781759 TI - A quantitative method for accounting human opinion, preferences and perceptions in ecosystem management. AB - Ecosystem management (EM) is a holistic approach, in which public participation in decision-making, and incorporation of human preferences, needs and perceptions in management plans is a main element. The decision-making from human opinion method (DeMHO), presented here, can be used in EM in selecting the more suitable and socially acceptable management plan, in order to protect or restore an ecosystem. The method focuses on the quantification of the human opinion, preferences and perceptions, which are investigated after research on the local population of the ecosystem. The results of this research are the inputs of the method; multi-criteria decision-making procedures, such as the analytic hierarchy process (AHP), the expected utility method (EUM) and compromise programming (CP) are used to assign the appropriate weights and rank according to their importance the interest groups, the issues to be studied, and the alternative management plans. The alternatives are also evaluated by assessing their sustainable character. The paper presents DeMHO and its application in the National Park of Eastern Macedonia and Thrace in Greece, after a research through a questionnaire on the local population. PMID- 12781760 TI - Effects of knowledge, personal attribution and perception of ecosystem health on depreciative behaviors in the intertidal zone of Pacific Rim National Park and Reserve. AB - Human activities levy a biological cost on ecosystems as resources are accessed and utilized at rates which are often incompatible with inherent ecosystem processes and structures. The recreational impact of humans upon intertidal zones and particularly fucoid algal assemblages is one major threat facing coastal ecosystems. The effect of human values, knowledge and perception in effecting biologically costly behaviors has rarely been examined. We hypothesize that with respect to intertidal zones: (1) Personal attribution and perception of ecosystem resiliency are more important than knowledge in determining the extent of depreciative behaviors individuals engage in, and; (2) Individuals who are uncertain about ecosystem resiliency will behave in a manner consistent with the 'precautionary principle'. We measured the depreciative behavior, and the attitudes and perceptions to ecosystem resilience, of visitors to Wick Headland in Pacific Rim National Park, British Columbia. Attitudes, knowledge, perceptions, and personal attribution were measured using questionnaire survey and structured interviews undertaken in situ. Depreciative behaviors of visitors were discreetly observed and correlated to the questionnaire survey and interview responses. We show that visitors who recorded greater knowledge of intertidal ecology engaged in more depreciative behaviors than visitors recording less knowledge. Visitors who perceived high ecosystem resilience in the intertidal zone engaged in significantly more behaviors eliciting biological cost than those who perceived low ecosystem resilience. Visitors who recorded uncertainty regarding ecosystem resilience engaged in significantly more depreciative behaviors than those who perceived low ecosystem resilience but slightly fewer depreciative behaviors than those who perceived high ecosystem resilience. Personal attribution was inversely correlated to the mean number of depreciative behaviors. We discuss the relevance of these results to the management of intertidal zones and marine protected areas, to multiple use management, the management of visitor impact, and natural resource use. PMID- 12781764 TI - Structural framework for catalysis and regulation in ribulose-1,5-bisphosphate carboxylase/oxygenase. AB - Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) is the enzyme assimilating CO2 in biology. Despite serious efforts, using many different methods, a detailed understanding of activity and regulation in Rubisco still eludes us. New results in X-ray crystallography may provide a structural framework on which to base experimental approaches for more detailed analyses of the function of Rubisco at the molecular level. This article gives a critical review of the field and summarizes recent results from structural studies of Rubisco. PMID- 12781765 TI - Role of the small subunit in ribulose-1,5-bisphosphate carboxylase/oxygenase. AB - Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) catalyzes the rate limiting step of CO2 fixation in photosynthesis, but O2 competes with CO2 for substrate ribulose 1,5-bisphosphate, leading to the loss of fixed carbon. Interest in genetically engineering improvements in carboxylation catalytic efficiency and CO2/O2 specificity has focused on the chloroplast-encoded large subunit because it contains the active site. However, there is another type of subunit in the holoenzyme of plants, which, like the large subunit, is present in eight copies. The role of these nuclear-encoded small subunits in Rubisco structure and function is poorly understood. Small subunits may have originated during evolution to concentrate large-subunit active sites, but the extensive divergence of structures among prokaryotes, algae, and land plants seems to indicate that small subunits have more-specialized functions. Furthermore, plants and green algae contain families of differentially expressed small subunits, raising the possibility that these subunits may regulate the structure or function of Rubisco. Studies of interspecific hybrid enzymes have indicated that small subunits are required for maximal catalysis and, in several cases, contribute to CO2/O2 specificity. Although small-subunit genetic engineering remains difficult in land plants, directed mutagenesis of cyanobacterial and green-algal genes has identified specific structural regions that influence catalytic efficiency and CO2/O2 specificity. It is thus apparent that small subunits will need to be taken into account as strategies are developed for creating better Rubisco enzymes. PMID- 12781766 TI - The life of ribulose 1,5-bisphosphate carboxylase/oxygenase--posttranslational facts and mysteries. AB - The life of ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco), from gene to protein to irreplaceable component of photosynthetic CO2 assimilation, has successfully served as a model for a number of essential cellular processes centered on protein chemistry and amino acid modifications. Once translated, the two subunits of Rubisco undergo a myriad of co- and posttranslational modifications accompanied by constant interactions with structurally modifying enzymes. Even after final assembly, the essential role played by Rubisco in photosynthetic CO2 assimilation is dependent on continuous conformation modifications by Rubisco activase. Rubisco is also continuously assaulted by various environmental factors, resulting in its turnover and degradation by processes that appear to be enhanced during plant senescence. PMID- 12781767 TI - Manipulating ribulose bisphosphate carboxylase/oxygenase in the chloroplasts of higher plants. AB - Transgenic manipulation of the photosynthetic CO2-fixing enzyme, ribulose bisphosphate carboxylase/oxygenase (Rubisco) in higher plants provides a very specific means of testing theories about photosynthesis and its regulation. It also encourages prospects for radically improving the efficiencies with which photosynthesis and plants use the basic resources of light, water, and nutrients. Manipulation was once limited to variation of the leaf's total content of Rubisco by transforming the nucleus with antisense genes directed at the small subunit. More recently, technology for transforming the small genome of the plastid of tobacco has enabled much more precise manipulation and replacement of the plastome-encoded large subunit. Engineered changes in Rubisco's properties in vivo are reflected as profound changes in the photosynthetic gas-exchange properties of the leaves and the growth requirements of the plants. Unpredictable expression of plastid transgenes and assembly requirements of some foreign Rubiscos that are not satisfied in higher-plant plastids provide challenges for future research. PMID- 12781768 TI - Phosphoenolpyruvate carboxylase: three-dimensional structure and molecular mechanisms. AB - Phosphoenolpyruvate carboxylase (PEPC; EC 4.1.1.31) catalyzes the irreversible carboxylation of phosphoenolpyruvate (PEP) to form oxaloacetate and Pi using Mg2+ or Mn2+ as a cofactor. PEPC plays a key role in photosynthesis by C4 and Crassulacean acid metabolism plants, in addition to its many anaplerotic functions. Recently, three-dimensional structures of PEPC from Escherichia coli and the C4 plant maize (Zea mays) were elucidated by X-ray crystallographic analysis. These structures reveal an overall square arrangement of the four identical subunits, making up a "dimer-of-dimers" and an eight-stranded beta barrel structure. At the C-terminal region of the beta barrel, the Mn2+ and a PEP analog interact with catalytically essential residues, confirmed by site-directed mutagenesis studies. At about 20A from the beta barrel, an allosteric inhibitor (aspartate) was found to be tightly bound to down-regulate the activity of the E. coli enzyme. In the case of maize C4-PEPC, the putative binding site for an allosteric activator (glucose 6-phosphate) was also revealed. Detailed comparison of the various structures of E. coli PEPC in its inactive state with maize PEPC in its active state shows that the relative orientations of the two subunits in the basal "dimer" are different, implicating an allosteric transition. Dynamic movements were observed for several loops due to the binding of either an allosteric inhibitor, a metal cofactor, a PEP analog, or a sulfate anion, indicating the functional significance of these mobile loops in catalysis and regulation. Information derived from these three-dimensional structures, combined with related biochemical studies, has established models for the reaction mechanism and allosteric regulation of this important C-fixing enzyme. PMID- 12781769 TI - Evolution of C4 phosphoenolpyruvate carboxylase. AB - C4 plants are known to be of polyphyletic origin and to have evolved independently several times during the evolution of angiosperms. This implies that the C4 isoform of phosphoenolpyruvate carboxylase (PEPC) originated from a nonphotosynthetic PEPC gene that was already present in the C3 ancestral species. To meet the special requirements of the C4 photosynthetic pathway the expression program of the C4 PEPC gene had to be changed to achieve a strong and selective expression in leaf mesophyll cells. In addition, the altered metabolite concentrations around C4 PEPC in the mesophyll cytoplasm necessitated changes in the enzyme's kinetic and regulatory properties. To obtain insight into the evolutionary steps involved in these altered enzyme characteristics, and even the order of these steps, the dicot genus Flaveria (Asteraceae) appears to be the experimental system of choice. Flaveria contains closely related C3, C3-C4, and C4 species that can be ordered by their gradual increase in C4 photosynthetic traits. The C4 PEPC of F. trinervia, which is encoded by the ppcA gene class, possesses typical kinetic and regulatory features of a C4-type PEPC. Its nearest neighbor is the orthologous ppcA gene of the C3 species F. pringlei. This latter gene encodes a typical nonphotosynthetic C3-type PEPC which is believed to be similar to the C3 ancestral PEPC. This pair of orthologous PEPCs has been used to map C4-specific molecular determinants for the kinetic and regulatory characteristics of C4 PEPCs. The most notable finding from these investigations was the identification of a C4 PEPC invariant site-specific mutation from alanine (C3) to serine (C4) at position 774 that was a necessary and late step in the evolution of C3 to C4 PEPC. The C3-C4 intermediate ppcA PEPCs are used to identify the sequence of events leading from a C3- to a C4-type PEPC. PMID- 12781770 TI - Control of the phosphorylation of phosphoenolpyruvate carboxylase in higher plants. AB - Phosphoenolpyruvate (PEP) carboxylase is regulated by reversible phosphorylation in higher plants. Recently several genes encoding PEP carboxylase kinase have been cloned. The purpose of this article is to assess the contribution that information on the structure and expression of these genes is making to our understanding of the posttranslational control of PEP carboxylase activity. PMID- 12781771 TI - Metabolic consequences of overproduction of phosphoenolpyruvate carboxylase in C3 plants. AB - Phosphoenolpyruvate carboxylase (PEPC) has a variety of functions in plants, including a major anaplerotic role in replenishing the tricarboxylic acid cycle with intermediates to meet the demand of carbon skeletons for synthesis of organic acids and amino acids. Various transgenic C3 plants that overproduce PEPC have been produced and analyzed in detail. The results indicate that foreign PEPC is under the control of the regulatory mechanisms intrinsic to the host plant and down-regulated so as not to cause detrimental metabolic effects, although the anaplerotic reaction is slightly enhanced by the foreign PEPC. By use of foreign PEPCs that can avert such regulation, metabolic flow is largely directed toward synthesis of organic acids and amino acids. Observations with transgenic C3 plants also shed light on the interrelation among various metabolic pathways inside the cell. PMID- 12781772 TI - Regulation and roles of phosphoenolpyruvate carboxykinase in plants. AB - Phosphoenolpyruvate carboxykinase (PCK) is probably ubiquitous in flowering plants, but is confined to certain cells or tissues. It is regulated by phosphorylation, which renders it less active by altering both its substrate affinities and its sensitivity to regulation by adenylates. In the leaves of some C4 plants, such as Panicum maximum, dephosphorylation increases its activity in the light. In other tissues such regulation probably avoids futile cycling between phosphoenolpyruvate and oxaloacetate. Although PCK generally acts as a decarboxylase in plants, its affinity for CO2 measured at physiological concentrations of metal ions is high and would allow it to be freely reversible in vivo. While its function in gluconeogenesis in seeds postgermination and in leaves of C4 and crassulacean acid metabolism plants is clearly established, the possible functions of PCK in other plant cells are discussed, drawing parallels with those in animals, including its integrated function in cataplerosis, nitrogen metabolism, pH regulation, and gluconeogenesis. PMID- 12781773 TI - Plant biotin-containing carboxylases. AB - Biotin-containing proteins are found in all forms of life, and they catalyze carboxylation, decarboxylation, or transcarboxylation reactions that are central to metabolism. In plants, five biotin-containing proteins have been characterized. Of these, four are catalysts, namely the two structurally distinct acetyl-CoA carboxylases (heteromeric and homomeric), 3-methylcrotonyl-CoA carboxylase and geranoyl-CoA carboxylase. In addition, plants contain a noncatalytic biotin protein that accumulates in seeds and is thought to play a role in storing biotin. Acetyl-CoA carboxylases generate two pools of malonyl CoA, one in plastids that is the precursor for de novo fatty acid biosynthesis and the other in the cytosol that is the precursor for fatty acid elongation and a large number of secondary metabolites. 3-Methylcrotonyl-CoA carboxylase catalyzes a reaction in the mitochondrial pathway for leucine catabolism. The exact metabolic function of geranoyl-CoA carboxylase is as yet unknown, but it may be involved in isoprenoid metabolism. This minireview summarizes the recent developments in our understanding of the structure, regulation, and metabolic functions of these proteins in plants. PMID- 12781774 TI - Combined effects of dynamic tissue shear deformation and insulin-like growth factor I on chondrocyte biosynthesis in cartilage explants. AB - Biophysical forces and biochemical factors play crucial roles in the maintenance of the integrity of articular cartilage. In this study, we explored the effect of dynamic tissue shear deformation and insulin-like growth factor I (IGF-I) on matrix synthesis by chondrocytes within native cartilage explants. Dynamic tissue shear in the range of 0.5-6% strain amplitude at 0.1 Hz was applied to cartilage explants cultured in serum-free medium. Dynamic tissue shear above 1.5% strain amplitude significantly stimulated protein and proteoglycan synthesis, by maximum values of 35 and 25%, respectively, over statically held control specimens. In the absence of tissue shear, IGF-I augmented protein and proteoglycan synthesis up to twofold at IGF-I concentrations in the range of 100-300 ng/ml. When tissue shear and IGF-I stimuli were combined, matrix biosynthesis levels were significantly higher than the maximal effect caused by either stimulus alone. However, there was no significant interaction between tissue shear and IGF-I as determined by two-way ANOVA. We then quantified the effect of dynamic tissue shear on the transport of IGF-I into and within cartilage explants. [125I]IGF-I was added to the medium, and the levels of intratissue [125I]IGF-I were directly measured as a function of time over 48 h in the presence and absence of continuous dynamic shear strain. Dynamic shear did not alter the rate of uptake of [125I]IGF-I into the explants, suggesting that convective diffusion of [125I]IGF-I is negligible under the shear strain conditions used. This is in marked contrast to the enhancement of transport reported in response to uniaxial dynamic compression. Taken together, these data suggest that (1) the stimulatory effect of tissue shear is via mechanotransduction pathways and not by facilitated transport of biochemical factors and (2) chondrocytes may possess complementary signal transduction pathways for biophysical and biochemical factors leading to changes in metabolic activity. PMID- 12781775 TI - The inhibition of degradation of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase by sterol regulatory element binding protein cleavage-activating protein requires four phenylalanine residues in span 6 of HMG-CoA reductase transmembrane domain. AB - 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) is the rate-limiting enzyme in the cholesterol biosynthetic pathway. This endoplasmic reticulum membrane protein contains a cytosolic catalytic domain and a transmembrane domain with eight membrane spans that are necessary for sterol-accelerated degradation. Competition experiments showed that wild-type transmembrane domains of HMGR and sterol regulatory element binding protein cleavage-activating protein (SCAP) blocked sterol-accelerated degradation of intact HMGR and HMGal, a model protein containing the membrane domain of HMGR linked to Escherichia coli beta galactosidase. However, mutant transmembrane domains of HMGR and SCAP whose sterol-sensing functions were abolished did not inhibit sterol-accelerated degradation of HMGR and HMGal. In addition, our mutagenesis studies on HMGal indicated that four Phe residues conserved in span 6 of HMGR and the sterol sensing domains of other sterol-related proteins are required for the regulated degradation of HMGR. These results suggest that HMGR and SCAP compete for binding to a sterol-regulated regulator protein, and this binding may need the four Phe residues. PMID- 12781776 TI - Identification of epitopes on cytochrome P450 3A4/5 recognized by monoclonal antibodies. AB - In this study we describe the mapping of epitopes on CYP3A4/5 recognized by a panel of monoclonal antibodies (MAbs). CYP3A4 and CYP3A5 cDNAs were cloned in GST expression vectors and the fusion proteins were subjected to Western blot. Eight MAbs reacted with the full-length GST-3A4 fusion protein as well as baculovirus cDNA-expressed CYP3A4, while six of these reacted with baculovirus cDNA-expressed CYP3A5. Five (MAb 347, 351, 352, 354, and 357) out of 8 MAbs were inhibitory in a metabolic assay using quinine as substrate. MAbs 352, 354, and 357 brought about a moderate inhibition of quinine metabolism (60-70%) while MAb 347 inhibited quinine 3- hydroxylation in human liver microsomes (n=6) by more than 70%. MAb 347 was a potent inhibitor of baculovirus-expressed CYP3A5-catalyzed metabolism of quinine (95%) at largescale sucker>northern pikeminnow>/=chinook salmon>/=redside shiner). At all supersaturation levels evaluated, percent of lateral line occlusion exhibited an inverse correlation to pore size but was not generally related to total dissolved gas level or time of exposure. This study suggests that the differences in lateral line pore diameters between species should be considered when using lateral line occlusion as an indicator of gas bubble trauma. PMID- 12781832 TI - Food choice by Blue-gray Tanagers in relation to protein content. AB - We tested discriminatory ability and food choice in relation to protein content of the diet in wild-caught Blue-gray Tanagers (Thraupis episcopus), a generalist tropical frugivorous bird. In two sets of experiments we offered to five individual birds in pair-wise choice trials two nearly iso-caloric experimental diets differing in their protein content only. Protein contents of the experimental diets were 4.6 vs. 1.4% in the first experiment and 3.2 and 1.5% (dry matter basis) in the second experiment. Response varied among individual tanagers, but 6 of the 10 birds showed a clear preference for the food highest in protein. Two individuals displayed a strong positional preference. When testing each treatment group, birds ate daily significantly more of the food that had higher protein content. We conclude that Blue-gray Tanagers prefer richer nitrogen foods. Our results also demonstrate that Blue-gray Tanagers have remarkable discriminatory abilities, they reacted to differences in protein content as small as 0.09% fresh matter. We show for the first time discriminatory ability and preference of wild frugivorous birds for foods richer in protein under controlled conditions. Our findings support the hypothesis that frugivorous birds can act as selective agents for fruit pulp composition. PMID- 12781833 TI - The fatty acid composition of oophagous tadpoles (Chirixalus eiffingeri) fed conspecific or chicken egg yolk. AB - We compared the lipid content and fatty acid composition of (1) the egg yolk of three anuran species (Chirixalus eiffingeri, Rhacophorus moltrechti and Buergeria robustus) and chicken eggs; and (2) C. eiffingeri tadpoles fed conspecific eggs or chicken egg yolk. Anuran and chicken egg yolk contained more non-polar than polar lipids but the proportions varied among species. Chicken egg yolk contained low amounts of 22:5n-3 in the polar lipid fraction, and B. robustus eggs did not contain any n-3 or n-6 non-polar lipids. The specific variation of lipid contents and fatty acid composition may relate to the maternal diet and/or breeding biology. In C. eiffingeri tadpoles that fed chicken yolk or frog egg yolk, the dominant components of polar and non-polar lipids were 16:0, 18:0, 18:1, and 18:2n-6, or 20:4n-6 fatty acids. C. eiffingeri eggs contained more n-3 fatty acids (e.g. 18:3n-3 and 20:5n-3) than chicken egg yolk, and tadpoles fed conspecific eggs contained more of these fatty acids than tadpoles fed chicken egg yolk. The compositional differences in the fatty acids between C. eiffingeri tadpoles that fed frog egg or chicken egg yolk are the reflection of the variation in the dietary sources. Our results suggest a direct incorporation of fatty acids into the body without or minimal modification, which provide an important insight into the physiological aspects of cannibalism. PMID- 12781834 TI - A symbiosome membrane is not required for the actions of two host signalling compounds regulating photosynthesis in symbiotic algae isolated from cnidarians. AB - In many cnidarians, symbiotic algae live within host-derived symbiosomes. We determined whether a symbiosome membrane alters the response of isolated symbiotic algae to two signalling compounds that regulate algal carbon metabolism. Host release factor (HRF), which stimulates photosynthate release, and photosynthesis inhibiting factor (PIF), which inhibits photosynthetic carbon fixation, are found in homogenised tissue of the scleractinian coral Plesiastrea versipora. Compared with seawater controls, photosynthate release from isolated algae incubated in P. versipora homogenate for 2 h in the light was: 6 to 19-fold higher from its own algae (free of symbiosomes); 19 to 32-fold higher from Zoanthid robustus algae (within symbiosomes) and 3 to 24-fold higher from Z. robustus algae (free of symbiosomes); and from cultured algae (free of symbiosomes) was seven-fold higher from Montipora verrucosa and four-fold higher from Cassiopeia xamachana. Incubation of algae in P. versipora homogenate inhibited photosynthesis by: 33-49% in P. versipora algae; 29-47% in Z. robustus algae (regardless of whether or not the symbiosome was present); and 25% in M. verrucosa algae. In C. xamachana algae, photosynthesis increased. We conclude that the symbiosome is not essential for, yet does not block, the effects of HRF and PIF. PMID- 12781835 TI - Metabolic priorities during starvation: enzyme sparing in liver and white muscle of Atlantic cod, Gadus morhua L. AB - Atlantic cod, Gadus morhua, respond to starvation first by mobilising hepatic lipids, then muscle and hepatic glycogen and finally muscle proteins. The dual role of proteins as functional elements and energetic reserves should lead to a temporal hierarchy of mobilisation where the nature of a function dictates its conservation during starvation. We examined (1) whether lysosomal and anti oxidant enzymes in liver and white muscle are spared during prolonged starvation, (2) whether the responses of these enzymes in muscle vary longitudinally. Hepatic contents of lysosomal proteases decreased with starvation, whereas those of catalase (CAT) increased and lysosomal enzymes of carbohydrate metabolism and glutathione S-transferase (GST) did not change. In white muscle, starvation decreased the specific activity of lysosomal enzymes of carbohydrate degradation and doubled that of cathepsin D (CaD). The activity of anti-oxidant enzymes and acid phosphatase in muscle was unchanged with starvation. In white muscle neither lysosomal enzymes nor anti-oxidant enzymes varied significantly with sampling position. In cod muscle, antioxidant enzymes, CaD and acid phosphatase are spared during a period of starvation that decreases lysosomal enzymes of carbohydrate metabolism and decreases glycolytic enzyme activities. In cod liver, the anti oxidant enzymes, CAT and GST, were also spared during starvation. PMID- 12781836 TI - Effect of ergotamine on plasma metabolite and insulin-like growth factor-1 concentrations in cows. AB - Bovine plasma was assayed to determine if ergotamine affected plasma metabolite and insulin-like growth factor-1 (IGF-1) concentrations. In Experiment 1, four cows received a single bolus intravenous injection of ergotamine tartrate (19 microg/kg body wt.) or saline vehicle in a crossover design 2 days after prostaglandin-induced luteolysis. Treatmentxtime affected plasma glucose, triglyceride, total cholesterol and IGF-1 concentrations. Glucose and cholesterol were increased after ergotamine. Triglycerides were elevated within 1 h after ergotamine, but were decreased 3 h after ergotamine treatment. Plasma IGF-1 decreased in response to ergotamine. Blood constituents were unchanged after treatment with saline. In Experiment 2, six cows received a single bolus intravenous injection of ergotamine (20 microg/kg body wt.) or saline vehicle in a crossover design 10 days after receiving norgestomet (6 mg) via subcutaneous ear implant. Treatmentxtime affected glucose, triglycerides, total cholesterol and IGF-1 concentrations. Glucose and cholesterol were increased after ergotamine. Triglycerides were elevated 1 h after ergotamine and decreased 3-7 h after ergotamine. Plasma IGF-1 decreased after ergotamine treatment. Blood constituents were unresponsive to the saline vehicle. Results indicated ergotamine altered plasma metabolite and IGF-1 concentrations in cows. PMID- 12781837 TI - Site-specific differences in the action of NRTI drugs on adipose tissue incubated in vitro with lymphoid cells, and their interaction with dietary lipids. AB - Existing theories of the origin of HIV-related adipose tissue redistribution syndrome cannot adequately explain simultaneous hypertrophy of certain depots and atrophy of others, or its occasional occurrence in untreated HIV infection. These experiments explore the hypothesis that hypertrophy of lymphoid tissue-containing adipose depots arises from drug-induced disruption to local interactions between perinodal adipocytes and activated lymphoid cells. Guinea pigs were fed on plain or lipid-supplemented (10% suet, sunflower or fish oil) chow ad libitum or restricted, and the popliteal lymph nodes were activated by repeated injection of lipopolysaccharide. Explants of perinodal and other samples from popliteal, mesentery, omentum and nodeless perirenal and epididymal depots were incubated with lymphoid cells and zidovudine, didanosine, lamivudine or stavudine at physiological concentrations (0.1-1 microg/ml) or interleukin-10 and interleukin 6, and basal and maximum lipolysis was measured. All drugs increased lipolysis from perinodal adipocytes, especially mesenteric, though less than exogenous cytokines. Effects on adipocytes from non-perinodal sites and nodeless depots were minimal. The sunflower-oil diet enhanced, and the fish-oil and restricted diets reduced, these effects. We conclude that these NRTI antiretroviral drugs modulate the local interactions between perinodal adipocytes and activated lymphoid cells. Local interactions, and hence the selective hypertrophy of node containing adipose depots, may be curtailed by dietary manipulation. PMID- 12781838 TI - Influence of 6-week exercise training on erythrocyte and liver antioxidant defense in hyperinsulinemic rats. AB - High dosage of fructose in rats causes insulin resistance and hyperinsulinemia. This study investigates the effect of physical exercise on oxidant-antioxidant balance in rats fed a high fructose diet, which show characteristic features of insulin resistance. Products of lipid peroxidation and the activity of enzymic antioxidants namely superoxide dismutase, catalase, glutathione peroxidase, glutathione-S-transferase and glutathione reductase, in red blood cells (RBCs) and liver were assayed. Levels of non-enzymic antioxidants alpha-tocopherol and ascorbic acid and of protein and non-protein thiols were also determined. The levels of lipid peroxides, diene conjugates, lipofuscin and hydroperoxides were significantly higher in the liver of fructose-fed rats. The RBCs showed significantly higher susceptibility to H(2)O(2)-induced stress compared to control rats. Inadequate antioxidant system was noted in high fructose-fed rats. Physical training to these rats reversed the adverse effects, which could be important in alleviating the pathological consequences of insulin resistance. PMID- 12781839 TI - Hepatic accumulation and effects of microcystin-LR on juvenile goldfish Carassius auratus L. AB - After intraperitoneal injection of microcystin-LR (MC-LR) (125 microg kg(-1) body wt.), the concentration of MC-LR in the liver of juvenile goldfish Carassius auratus (30 g body wt.) was assayed by a modified protein phosphatase inhibition method. A temporary accumulation occurred from 3 to 48 h post-injection, followed by a significant decrease between 48 and 96 h. Under our experimental conditions, contamination by MC-LR did not change ionic homeostasis, as attested by blood osmolality values and gill Na(+)/K(+) ATPase activity. Light microscopy observations revealed lesions and cellular necrosis progression, which was concomitant with an increase in enzyme activity of plasma aspartate aminotransferase (AspAT), alanine aminotransferase (AlaAT) and L-lactate dehydrogenase (LDH) and with a decrease of hepatic glutathione-S-transferase (GST) activity. Structural alterations and enzymatic activity modifications became significant within 24 h post-injection. Recovery of hepatocytes on day 21 after MC-LR injection was evident, together with a decrease in the MC-LR equivalent content of the liver. PMID- 12781841 TI - Increased superoxide dismutase and Na+, K+-ATPase activities in aortic strips from potassium-adapted rats: implication for altered vascular reactivity. AB - The contributions of superoxide dismutase (SOD) and Na(+), K(+)-ATPase to the altered vascular reactivity in potassium-adapted rats were investigated to test the hypothesis that smooth muscle hyperpolarisation may be involved. Isometric contractions to noradrenaline (NA), 5-hydroxytryptamine (5-HT), and relaxations to acetylcholine (ACh), levcromakalim (LEV) and sodium nitroprusside (SNP), were measured in aortic rings from potassium-adapted rats. Pieces of the aortae were also excised from the animals and assayed for SOD and Na(+), K(+)-ATPase. Maximum contractile responses were significantly attenuated (P<0.05) in aortic rings from the potassium-adapted rats to NA and 5-HT, while relaxations were also significantly augmented (P<0.05) in the same rings to LEV and SNP, but not to ACh. Both SOD and Na(+), K(+)-ATPase activities were significantly higher (P<0.05) in the aortae from the potassium-adapted rats compared to controls. It is concluded that the alteration in vascular smooth muscle reactivity may be due to hyperpolarisation caused by the activities of SOD and Na(+), K(+)-ATPase. PMID- 12781840 TI - The role of abiotic factors and pesticide levels on enzymatic activity in the freshwater mussel Anodonta cygnea at three different exposure sites. AB - Natural variation in abiotic factors, such as temperature and pH, probably influence the activity of enzymes used as potential biomarkers in bivalve mollusks to assess environmental contamination in the field. Changes in levels of an enzymatic biomarker may thus merely reflect natural variation in the annual physiological cycle of a species rather than exposure to contaminants. To investigate this issue, we documented the relationship between pesticide levels in water and three different enzymatic biomarkers over 1 year in enclosed populations of the freshwater unionid mussel Anodonta cygnea at three different sites of exposure. We considered the natural variation in temperature, pH and dissolved oxygen over the year and across the different sites as a potential correlate of enzymatic activity to disentangle the relative contribution of abiotic factors and pesticide levels. Pesticide levels varied among the three sites and over the course of the year. Catalase (CAT) and acetylcholinesterase activity (AChE) varied as a function of abiotic factors but showed no relation to pesticide levels. Glutathione S-transferase (GST) activity was also related to abiotic factors but also decreased with increases in total pesticide levels. The lack of activity induction or inhibition by pesticides and the natural variation in abiotic factors among sites and across time limits the use of CAT and AChE to assess environmental contamination in this species. PMID- 12781842 TI - Effect of microcystin on ion regulation and antioxidant system in gills of the estuarine crab Chasmagnathus granulatus (Decapoda, Grapsidae). AB - The objective of this work was to evaluate mechanisms of microcystin toxicity on crustacean species. Adult male crabs of Chasmagnathus granulatus (13.97+/-0.35 g) acclimated to low salinity (2 per thousand ) were injected with saline (control) or Microcystis aeruginosa aqueous extract (39.2 microg/l) at 24 h intervals for 48 h. After the exposure period, the anterior and posterior gills were dissected, measuring Na(+),K(+)-ATPase and glutathione-S-transferase (GST) activity. Total oxyradical scavenging capacity (TOSC) and lipid peroxides (LPO) content were also determined. Na(+),K(+)-ATPase activity in anterior gills was significantly lower in crabs injected with toxin than in control crabs, while no significant difference in the enzyme activity was detected in posterior gills. Both sodium and chloride concentration in the hemolymph were not affected by toxin exposure. Significant changes in GST activity were detected in posterior gills, with higher values being observed in the toxin-injected crabs. Crabs exposed to microcystin also showed a significant increase in the TOSC value against peroxyl radicals, for both anterior and posterior gills. Lipid peroxides level did not change in both gill types after exposure to the toxin. The increased levels of TOSC suggest the occurrence of a crab response against oxidative stress induced by toxin injection, which prevents lipid peroxidation. PMID- 12781843 TI - Alterations in cell membrane properties caused by perfluorinated compounds. AB - The recent detection of perfluorinated compounds (PFCs) in wildlife from even remote locations has spurred interest in the environmental occurrence and effects of these chemicals. While the global distribution of PFCs is increasingly understood, there is still little information available on their effects on wildlife. The amphiphillic nature of PFCs suggests that their effects could be primarily on cell membranes. In this study we measured the effects of PFCs on membrane fluidity and mitochondrial membrane potential using flow cytometry and effects on membrane permeability using cell bioassay procedures (H4IIE, MCF-7, PLHC-1). Of the PFCs tested, only perfluorooctane sulfonic acid (PFOS) increased the permeability of cell membranes to the hydrophobic ligands used. Three PFCs were tested in the membrane fluidity assay: PFOS, perfluorohexane sulfonic acid (PFHS), and perfluorobutane sulfonic acid (PFBS). PFOS increased membrane fluidity in fish leukocytes in a dose-dependent fashion, while PFHS and PFBS had no effect in the concentration range tested. The lowest effective concentrations for the membrane fluidity effects of PFOS were 5-15 mg/l. Effects on mitochondrial membrane potential occurred in the same concentration range as effects on membrane fluidity. This suggests that PFOS effects membrane properties at concentrations below those associated with other adverse effects. PMID- 12781844 TI - Reactive oxygen species in pregnant rats: effects of exercise and thermal stress. AB - With the aim of evaluating the effect of interaction between physical training or exercise only during pregnancy and thermal stress on oxidative stress, and antioxidant mechanism sedentary pregnant rats (PS), exercised pregnant rats only during pregnancy (PE) and trained rats submitted to also exercise during pregnancy (PT) were compared (N=63). Exercise sessions consisted of swimming at 80% of maximal work load supported into water at 28 degrees C (hypothermia, PS 28, PE28, PT28) or 35 degrees C (thermal neutrality, PS35, PE35, PT35) or 39 degrees C (hyperthermia, PS39, PE39, PT39), for 30 min. The initial body weight in all groups of rats was from 177 to 207 g. On the 20th day of pregnancy, 24 h after the last immersion or swimming session venous blood was collected to determine oxidative stress. Plasma concentrations of means malondialdehyde (MDA) values measured as thiobarbituric acid reactive substances (TBARS); total glutathione (GSH) and vitamin E were determined. The oxidative stress index was calculated from the ratio TBARS/GSH and TBARS/Vitamin E. TBARS did not change on the group PE at different temperatures of water; TBARS were higher for PS28 than PS35 and PS39; PT35 had higher values than PT28 and PT39. For GSH, PS39 was lower than PS35; PE28 was higher than PE35 and PE39 and PT35 were lower than PT28 and PT39. Plasma concentration of vitamin E did not present any difference for sedentary rats at different water temperatures, but for PE28, the values were lower than for PE35 and PE39, whereas PT39 was lower than PT35 and PT28. In relation to TBARS/GSH, it was verified an increase in oxidative stress for PS28 (in relation to PS35 and PS39), PE35, and PT35 (in relation to PE28 and PE39 or PT28 and PT39); regarding the ratio TBARS/vitamin E, the highest values were obtained at 35 degrees C for PS and PT groups and at 39 for PE group. These results have shown the great complexity of the interaction between physical training, thermal stress and pregnancy. Apparently, hypothermia produces large index of oxidative stress only in sedentary rats, but this index was greater at 35 degrees C in relation to extreme temperatures for trained rats. These results have suggested that physical training allows a more efficient activation of antioxidant mechanisms under thermal stress. PMID- 12781845 TI - Harmonization of anthropometric measurements for a multicenter nutrition survey in Spanish adolescents. AB - OBJECTIVE: Although the need for accurate anthropometric measurement has been repeatedly stressed, reports on growth and physical measurements in human populations rarely include estimates of measurement error. We describe the standardization process and reliability of anthropometric measurements carried out in a pilot study. METHODS: For the intraobserver assessment of anthropometric measurements, we studied 101 adolescents (58 boys and 43 girls) from five cities. For interobserver assessment, we studied 10 adolescents from the same class in Zaragoza and different from those in the intraobserver sample. RESULTS: For skinfold thickness, intraobserver technical errors of measurement (TEMs) in general were smaller than 1 mm; for circumferences, TEMs in general were smaller than 1 cm. Intraobserver reliability for skinfold thickness was greater than 95% for almost all cases; for circumferences, intraobserver reliability generally was greater than 95%. Interobserver TEMs ranged from 1 to 2 mm for the six skinfold thicknesses measured; for circumferences, TEMs were smaller than 1 cm for the arm, biceps, and waist and between 1 and 2 cm for the hip and thigh. Interobserver reliabilities for skinfold thickness and circumference were always greater than 90%, except for biceps skinfold. CONCLUSIONS: Our results are in agreement with those recommended in the literature. Therefore, these anthropometric measures seem to be adequate to assess body composition in a multicenter survey in adolescents. PMID- 12781846 TI - Postoperative enteral stimulation by gut feeding improves outcomes in severe acute pancreatitis. AB - OBJECTIVE: We assessed the clinical effectiveness of postoperative enteral stimulation by gut feeding in patients with severe acute pancreatitis (SAP). METHODS: Medical records of 63 patients who were operated on within the past 4 y due to deterioration of SAP were included in this retrospective study. Patients were stratified in gut feeding (GF; n = 33) and standard therapy (ST; n = 30) groups according to the postoperative therapy provided. The GF group received postoperative standard therapy and enteral stimulation by gut feeding, and the ST group received standard therapy only. The Acute Physiology and Chronic Health Evaluation II score, incidence of the systemic inflammatory response syndrome (SIRS), multiple organ dysfunction syndrome (MODS), daily calories supply, complication rate, and outcomes were analyzed. RESULTS: Patient characteristics did not differ between groups when considering age and severity of the disease. All patients underwent similar surgical interventions. SIRS and MODS were observed initially with the same frequency in both groups. Regression of MODS and a lower postoperative complication rate was observed more often in the GF group. Development of early pulmonary complications was observed in 12.1% to 13.3% in both groups, irrespective of the time of surgery. Subsequently, pulmonary complications developed in 15.2% of GF patients compared with 43.3% of ST patients (P <0.05). Acute renal insufficiency developed similarly in 33.3% of the GF patients and in 26.7% of the ST patients within 3 d after admission. Acute renal insufficiency developed later on only in the ST group (26.7%, P <0.05). Wound- and catheter-related septic complications were considerably more frequent in the ST group (30.0%) than in the GF group (9.1%, P <0.05). Intensive care and hospital stays did not differ. Postoperative gut feeding was associated with 6.1% mortality in the GF compared with 26.7% in the ST (P <0.05). CONCLUSIONS: Enteral stimulation by gut feeding is an effective supplement in the postoperative therapy of patients with SAP. PMID- 12781847 TI - Bioelectrical impedance vector distribution in the first year of life. AB - OBJECTIVE: We assessed the bioelectrical impedance vector distribution in a sample of healthy infants in the first year of life, which is not available in literature. METHODS: The study was conducted as a cross-sectional study in 153 healthy Caucasian infants (90 male and 63 female) younger than 1 y, born at full term, adequate for gestational age, free from chronic diseases or growth problems, and not feverish. Z scores for weight, length, cranial circumference, and body mass index for the study population were within the range of +/-1.5 standard deviations according to the Euro-Growth Study references. Concurrent anthropometrics (weight, length, and cranial circumference), body mass index, and bioelectrical impedance (resistance and reactance) measurements were made by the same operator. Whole-body (hand to foot) tetrapolar measurements were performed with a single-frequency (50 kHz), phase-sensitive impedance analyzer. The study population was subdivided into three classes of age for statistical analysis: 0 to 3.99 mo, 4 to 7.99 mo, and 8 to 11.99 mo. Using the bivariate normal distribution of resistance and reactance components standardized by the infant's length, the bivariate 95% confidence limits for the mean impedance vector separated by sex and age groups were calculated and plotted. Further, the bivariate 95%, 75%, and 50% tolerance intervals for individual vector measurements in the first year of life were plotted. RESULTS: Resistance and reactance values often fluctuated during the first year of life, particularly as raw measurements (without normalization by subject's length). However, 95% confidence ellipses of mean vectors from the three age groups overlapped each other, as did confidence ellipses by sex for each age class, indicating no significant vector migration during the first year of life. CONCLUSIONS: We obtained an estimate of mean impedance vector in a sample of healthy infants in the first year of life and calculated the bivariate values for an individual vector (95%, 75%, and 50% tolerance ellipses). PMID- 12781848 TI - Folate, vitamin B12, and homocysteine status. findings of no relation between human papillomavirus persistence and cervical dysplasia. AB - OBJECTIVES: Human papillomavirus (HPV) infections are the cause of most, if not all, cervical cancers. Women consistently positive for oncogenic type HPV infections have a greater risk of developing cervical dysplasia compared with women transiently infected. HPV infection alone appears to be insufficient to produce disease, suggesting that other cofactors may be needed. Folate, vitamin B12, and homocysteine, through their role in DNA methylation, may be involved in cervical neoplasia. METHODS: This study examined the associations between HPV persistence and circulating folate, vitamin B12, and homocysteine levels among 91 low-income Hispanic women. Further, the relation of these nutrients to cervical pathology was evaluated. HPV status was determined at two visits approximately 3 mo apart. RESULTS: Adjusted mean circulating concentrations of folate, vitamin B12, and homocysteine were not statistically different between women with two positive HPV tests, one positive test, or two negative HPV tests. No association was observed between tertiles of folate, vitamin B12, or homocysteine and HPV persistence risk. Further, adjusted mean levels of these nutrients were not statistically different between cytologic grades. CONCLUSIONS: Results from this small study did not support a role for folate, vitamin B12, or homocysteine in HPV persistence or cervical dysplasia. PMID- 12781849 TI - Non-obese (body mass index < 25 kg/m2) Asian Indians with normal waist circumference have high cardiovascular risk. AB - OBJECTIVE: Although the prevalence of obesity is not high in Asian Indians, increased prevalence rates of metabolic perturbations and cardiovascular risk factors have been reported. In this study, we evaluated body mass index (BMI), anthropometric measurements, and body fat profiles of obese and non-obese subjects and correlated those values with cardiovascular risk factors. METHODS: This cross-sectional study involved 639 subjects (170 men and 469 women) from low socioeconomic stratum residing in urban slums of New Delhi. Non-obese subjects were categorized into quartiles of percentage of body fat (%BF) and waist circumference (WC). Using logistic regression analysis, the odds ratios (ORs) and 95% confidence intervals (CIs) for the occurrence of cardiovascular risk factors (diabetes mellitus [DM], hypertension, hypercholesterolemia, hypertriglyceridemia, low levels of high-density lipoprotein cholesterol, and high levels of low-density lipoprotein cholesterol) were computed with first quartile in the non-obese group as the reference category for the following five categories: %BF and WC quartiles in the non-obese group and obese group (BMI > or = 25 kg/m(2)). RESULTS: In the non-obese group, approximately 66% of men and 88% of women had at least one risk factor. In non-obese women, significantly high ORs and 95% CIs were found for hypertension (3.2, 1.4-7.2), DM (2.8, 1.2-6.3), and hypertriglyceridemia (3.9, 1.9-8.3) in the upper quartiles of %BF, and significantly high OR (95% CIs) was observed for hypertriglyceridemia (3.9, 1.4 10.8) in non-obese men. Among non-obese men, significantly high OR (95% CIs) was found for hypertriglyceridemia (3.2, 1.0-10.3); among non-obese women, significantly high ORs (95% CIs) were observed for hypertension (2.5, 1.1-5.8) and hypertriglyceridemia (2.5, 1.2-5.2) in the normal range of WC (70-80 cm). CONCLUSION: The study showed that Asian Indians have excess cardiovascular risk at BMI and WC values considered "normal." These data suggested that definitions of "normal" ranges of BMI and WC need to be revised for Asian Indians. PMID- 12781850 TI - Bioimpedance vector pattern in cancer patients without disease versus locally advanced or disseminated disease. AB - OBJECTIVE: Bioelectrical impedance vector analysis allows non-invasive evaluation of soft tissue hydration and mass through pattern analysis of vector plots as height, normalized resistance, and reactance measurements. METHODS: Whole-body impedance measurements were made with a single frequency (50 kHz) analyzer (BIA 101, Akern/RJL Systems) in 148 adult, white, male subjects 45 to 85 y old: 56 healthy control subjects, 31 cancer patients after surgical procedure (without disease), and 61 patients with locally advanced (30 patients) or disseminated (31 patients) disease with the same body mass index and age. All patients were free from antineoplastic treatment and active nutritional intervention. RESULTS: Mean vectors of cancer groups without disease and locally advance disease versus the control group were characterized by a comparable normalized resistance component with a reduced reactance component (separate 95% confidence limits, P < 0.05), indicating a comparable ionic conduction (hydration) with loss of dielectric mass (cell membranes and tissue interfaces) of soft tissues. Overlapping 95% confidence limits of their mean vectors indicated comparable electrical tissue properties in less versus more advanced disease. CONCLUSION: Monitoring vector displacement trajectory toward the reference target vector position may represent useful feedback in support therapy planning of individual patients. PMID- 12781851 TI - Evaluation of nutritional practice in hospitalized cirrhotic patients: results of a prospective study. AB - OBJECTIVE: We evaluated the impact of malnutrition and nutrition practice in 396 hospitalized cirrhotic patients (Child stages A, B, and C: 60, 169, and 167 patients) in a readaptative unit for liver diseases. METHODS: Triceps skinfold thickness and mid-arm muscle circumference were measured at admission, and spontaneous dietary intake was evaluated at admission and during hospital stay. RESULTS: Ascites was associated with impairment of nutrition status: 49% and 30.4% of non-ascitic patients, 49.1% and 40.5% of patients with mild ascites, and 65.5% and 48.3% of patients with tense ascites had mid-arm muscle circumferences and triceps skinfold thicknesses, respectively, below the fifth percentile of a reference population (P < 0.05 and P = 0.02, respectively). Decrease in dietary intake paralleled worsening of liver failure: 48% and 34% of Child A patients, 51.7% and 35.8% of Child B patients, and 80.3% and 62.9% of Child C patients at admission had caloric intakes below 30 kcal/kg of body weight and protein intakes below 1 g/kg of body weight, respectively (P < 0.001). Mortality rate was 15.4% during hospital stay; Child-Pugh score (P = 0.0014), age (P = 0.0029), severe septic complications (P = 0.0050), and decrease in caloric intake during hospital stay (P = 0.0072) were independently associated with mortality. Twenty-four patients received enteral feeding that was initiated before admission in four patients and after 12.4 +/- 8.3 d of hospitalization in 20 patients because of low caloric intake (<25 kcal/kg) despite oral supplements. Patients receiving enteral feeding were older (P < 0.01), had a higher Child-Pugh score (P < 0.01), and a higher mortality rate (P < 0.001) than other patients. CONCLUSIONS: Hospitalized cirrhotic patients have a high prevalence rate of malnutrition, and most do not satisfy their nutritional requirements. Decrease in caloric intake is an independent risk factor of short-term mortality. Enteral nutrition after failure of oral supplementation has no clinical benefit. Tube feeding may be indicated earlier in the course of the disease. PMID- 12781852 TI - Effect of folic acid supplementation on plasma zinc concentrations of young women. AB - OBJECTIVE: Women of reproductive age are advised to consume supplements or fortified foods containing at least 400 microg/d folic acid for the prevention of neural tubes defects. Concerns exist about the adverse effects of folic acid on zinc status. METHODS: Seventy-eight women (18 to 49 y) were assigned for 12-wk to receive either a placebo or a 400 microg/d folic acid supplement. RESULTS: At 12 wk mean (95% CI) red cell folate increased by 431 (350-511) nmol/L in the supplemented group relative to placebo (P < 0.001) but there was no change in plasma zinc in either group (P = 0.213). CONCLUSIONS: Folic acid supplementation does not reduce plasma zinc concentrations in women of childbearing age. PMID- 12781853 TI - Making bread with sourdough improves mineral bioavailability from reconstituted whole wheat flour in rats. AB - OBJECTIVE: We compared the effects of different kinds of bread fermentation on mineral bioavailability. METHODS: Wistar rats were fed one of the following experimental diets for 21 d: control, reconstituted whole wheat flour (white flour plus bran), yeast bread, and sourdough bread. The apparent mineral absorption and intestinal fermentation were measured in each animal. RESULTS: Phytate contents in yeast and sourdough bread were lower than in reconstituted whole wheat flour (-52% and -71%, respectively). Total cecal pool of short-chain fatty acids, in particular the butyrate pool, was significantly increased by the ingestion of unrefined products. Calcium homeostasis was not modified by these nutritional conditions, whereas magnesium absorption was significantly greater in rats fed the control and sourdough diets than in those consuming whole wheat flour and yeast bread. Magnesium kidney excretion was slightly stimulated by sourdough bread. Compared with the control diet, iron balance was significantly reduced by reconstituted whole wheat flour diet. Yeast bread making counteracted the deleterious effects of whole wheat on iron absorption, whereas sourdough bread making enhanced iron absorption. Further, liver and plasma iron and transferrin saturation levels were lower in rats adapted to the flour diet than in other groups. Zinc absorption was strongly depressed in the presence of unprocessed reconstituted whole wheat flour in the diet, but yeast fermentation afforded a zinc assimilation comparable to the control diet, whereas the sourdough bread led to maximal zinc absorption. Copper absorption increased significantly when rats were fed the sourdough bread, whereas unprocessed whole flour depressed copper absorption (-41% versus control diet). CONCLUSION: Mineral bioavailability from reconstituted whole wheat flour can be improved by bread making. Although yeast fermentation minimizes the unfavorable effects of phytic acid, sourdough bread is a better source of available minerals, especially magnesium, iron, and zinc. PMID- 12781854 TI - In vitro effects of selenite and mercuric chloride on liver thiobarbituric acid reactive substances and non-protein thiols from rats: influences of dietary cholesterol and polyunsaturated and saturated fatty acids. AB - OBJECTIVE: We measured the in vitro effects of mercuric chloride (Hg2+) and selenite (Se4+) on hepatic 2-thiobarbituric acid-reactive substances (TBARS) and non-protein sulfhydryl (NPSH) levels of rats fed diets enriched with polyunsaturated or saturated fatty acids with and without cholesterol. METHODS: Male Wistar rats (21 d old) were assigned to one of four groups and fed diets containing 20% soybean oil, 20% soybean oil plus 1% cholesterol, 20% coconut oil, or coconut oil plus 1% cholesterol. After the feeding period (6 wk), body weight gain was equal in all groups. TBARS levels and NPSH content were measured after in vitro exposure to mercuric chloride (100 microM) and sodium selenite (25 microM) for 1 h. RESULTS: The lipid peroxidation, measured as TBARS levels in the control group, were statistically higher in hepatic homogenates of rats fed diets containing soybean oil than in groups fed coconut oil (P = 0.009). However, cholesterol supplementation did not change TBARS levels. Selenite alone did not modify TBARS production, whereas mercury alone significantly increased TBARS levels. Moreover, Se4+ protected against mercury-induced lipid peroxidation only in rats fed diets containing coconut oil. In the control group, dietary fat acids did not change NPSH levels. Selenite produced higher oxidative effects toward NPSH content, whereas Hg2+ decreased NPSH levels only in liver from rats fed diets containing soybean oil. NPSH levels were higher after concomitant exposure to Se4+ and Hg2+ chloride that after exposure to Se4+ alone, suggesting an interaction between Hg2+ and Se4+. Catalase activity was higher in animals fed diets containing soybean oil. Dietary cholesterol decreased glutathione peroxidase activity. CONCLUSION: Together these results indicated that the protective effect of Se4+ against mercury-induced lipid peroxidation depends on dietary fat saturation. PMID- 12781855 TI - Effect of green tea in the prevention and reversal of fasting-induced intestinal mucosal damage. AB - OBJECTIVE: Epidemiologic studies have suggested that high consumption of green tea protects against the development of chronic active gastritis and decreases the risk of stomach cancer. The effect of green tea on the intestinal mucosa was not studied previously, so we examined the effects of green tea on the intestinal mucosa of fasting rats in a controlled experimental setting. METHODS: Two sets of experiments were performed. In the recovery set, rats were fasted for 3 d, after which they were allowed free access to water, black tea, green tea, or vitamin E for 7 d. On day 8, the animals were killed, and small bowels were removed for histologic examination. In the pretreatment set, rats were allowed a normal diet, but the water supply was replaced with green tea, black tea, or vitamin E for 14 d. They were subsequently fasted for 3 d. On day 4, the rats were killed, and small bowels were removed for histologic examination. RESULTS: In the recovery set, fasting for 3 d caused shortening of villi, atrophy, and fragmentation of mucosal villous architecture, with a significant (P < 0.0001) reduction in the length and surface area of the villi. Ingestion of green tea and, to a lesser extent, vitamin E for 7 d helped in the recovery of villi to normal. In the pretreatment set, drinking green tea, black tea, or vitamin E for 14 d before fasting protected intestinal mucosa from damage. CONCLUSION: The mucosal and villous atrophy induced by fasting was reverted to normal by the ingestion of green tea and, to a lesser extent, vitamin E. Black tea ingestion had no effect. In addition, ingestion of black tea, green tea, and vitamin E before fasting protected the intestinal mucosa against atrophy. PMID- 12781856 TI - Detection of segmental internal fat by bioelectrical impedance analysis in a biological phantom. AB - OBJECTIVE: Quantification of internal adipose tissue such as visceral adipose tissue currently relies on expensive, cross-sectional imaging modalities. The purpose of this study was to test the hypothesis that surface impedance, determined by bioimpedance analysis, might be used to predict regional internal fat content change in a phantom model. METHODS: Fresh hollowed-out cucumbers were used as cylindrical biological phantoms to test this hypothesis. After removal of the seeds, the cucumbers were filled with normal saline, mixture of saline and corn oil, or porcine adipose tissue bathed in saline. Surface resistance and reactance were measured with a bioimpedance analyzer accurate to 0.1 Omega (Quantum 10X, RJL Systems), and impedance was calculated. A linear regression model was used to interpret the association between composition and impedance. RESULTS: Surface impedance varied linearly with changes in the relative internal corn oil portions (r- = 0.98). A similar relation was noted with porcine adipose tissue bathed in saline (r(2) = 0.95) regardless of the specific position of adipose tissue within the cucumber. CONCLUSION: Surface impedance measured by bioimpedance analysis can detect variations in fat content in the interior of a cylindrical phantom. PMID- 12781857 TI - Malondialdehyde production in Jurkat T cells subjected to oxidative stress. AB - OBJECTIVE: We investigated the relation between membrane lipid peroxidation, as evaluated by malondialdehyde (MDA), and oxidative stimuli in the Jurkat T-cell line and designed a cellular model to assess the antioxidant potential of compounds. METHODS: Jurkat T cells were subjected to different concentrations of Fe(2+) ions (from 25 to 150 micromol/L) or H(2)O(2) (from 0.1 to 5 mmol/L), and MDA was determined after separation in high-performance liquid chromatography of the adduct with thiobarbituric acid. MDA production also was investigated in cells supplemented with epigallocatechin gallate and genistein and subjected to Fe(2+) oxidative treatment. RESULTS: MDA production increased with the concentration of Fe(2+), whereas H(2)O(2) had no effect at any concentration. Oxidative stress for 15 min or 2 h produced similar MDA levels. The supplementation of epigallocatechin gallate partly prevented MDA production (about 40%, P < 0.05), whereas genistein exerted no preventive effect on lipid peroxidation. CONCLUSION: We propose this cellular model, consisting of Jurkat T cells subjected to 100 micromol/L of Fe(2+) for 15 min, to study the protective effect of antioxidant supplementation against membrane lipid peroxidation. PMID- 12781858 TI - Partially hydrolyzed guar gum: clinical nutrition uses. AB - OBJECTIVE: This paper provides a review of research on partially hydrolyzed guar gum that is relevant to clinical nutrition practice. METHODS: All relevant papers published on partially hydrolyzed guar gum were reviewed and the results summarized. RESULTS: Partially hydrolyzed guar gum (PHGG) is a water-soluble dietary fiber with a wide range of uses in clinical nutrition. Its low viscosity allows its use in enteral products and beverages. PHGG can be added to enteral formulas and food products as a dietary fiber source. PHGG provides the benefits associated with dietary fiber ingestion. Addition of PHGG to the diet reduced laxative dependence in a nursing home population. PHGG also reduced the incidence of diarrhea in septic patients receiving total enteral nutrition and reduced symptoms of irritable bowel syndrome. PHGG also increased production of Bifidobacterium in the gut. CONCLUSION: The ease of use of PHGG and its clinical effectiveness make it a good choice in clinical nutrition practice. PMID- 12781859 TI - Elevated plasma cholecystokinin and appetitive ratings after consumption of a liquid meal in humans. AB - OBJECTIVE: This study had two objectives. The first was to evaluate the possibility that, in a previous study, a soup preload augmented the reduction of food intake in a test meal induced by an exogenous infusion of cholecystokinin (CCK) because the soup also endogenously released CCK. The second was to compare CCK release by soup between men and women to determine whether the increased satiating effectiveness of soup in women as opposed to men could have been partly attributable to differences in CCK release. METHODS: By using a bioassay that measures all of its known isoforms, we determined plasma CCK levels at baseline and at several times postprandially in eight healthy, non-obese men and women (four of each sex). Each subject ingested 800 g of tomato soup, which was followed 30 min later by 300 g of a yogurt shake. Appetitive ratings were also collected and related to CCK levels. RESULTS: Ingestion of tomato soup significantly increased plasma CCK levels by 3.81 pmol/L (+/- 1.21 standard error, P = 0.016) over baseline within 30 min in all subjects combined. When CCK concentrations at 5 min after soup and 5 min after yogurt were averaged, the women's mean averaged concentration was 5.58 pmol/L (+/- 1.994, t = 2.80, P = 0.0073) higher than the men's. The elevated levels persisted but did not rise further upon consumption of the yogurt shake. Hunger ratings declined and fullness ratings increased after eating, although patterns of ratings did not match exactly patterns of CCK release. CONCLUSIONS: A large quantity of tomato soup stimulates significant CCK release; therefore, some of the satiating effects of soup preloads could have been mediated by an elevation in endogenous CCK. PMID- 12781860 TI - Reference BIA data in neonates and young infants. PMID- 12781861 TI - Fat patterning: indicators and implications. PMID- 12781862 TI - Pathobiology of metabolic syndrome X in obese and non-obese South Asian Indians: further discussion and some suggestions. PMID- 12781864 TI - Nutrition, saliva, and oral health. PMID- 12781863 TI - Nutrition support for the patient with an open abdomen after major abdominal trauma. AB - OBJECTIVE: Nutrition support in the severely injured trauma patient is crucial to minimize the hypermetabolic stress response. Even though enteral nutrition is the preferred method of feeding, it is not always feasible after multiple trauma. We present a complex nutritional case in a patient who sustained severe abdominal trauma with a severe liver injury, rib fractures, and pulmonary contusion. METHODS: The patient required several repeat laparotomies, abdominal packing, and temporary abdominal closure. The clinical course was complicated by hypotension requiring multiple vasopressors; coagulopathy requiring more than 35 U of packed red cells, more than 50 U of fresh frozen plasma, and more than 80 U of platelets; acute renal failure requiring dialysis; and pneumonia and acute respiratory distress syndrome requiring intricate ventilator management. Nutrition intervention began on post-trauma day 4 with total parenteral nutrition due to hypotension, resuscitation, and massive bowel edema; by post-trauma day 8. the patient was receiving goal nutrients. RESULTS: On post-trauma day 27, bowel edema was significantly less, and a nasoenteric feeding tube was placed and enteral feeding initiated. By post-trauma day 31, full enteral feeds were tolerated, and total parenteral nutrition was stopped. Nutrient provision was adjusted daily to account for organ and metabolic changes including hepatic, pulmonary, and renal dysfunction. The patient did well and was eventually extubated and eating a regular diet. CONCLUSION: With careful monitoring and adjusting of the nutritional plan, a hypermetabolic complex trauma patient with an open abdomen can be fed optimally, safely, and successfully despite increased bowel edema and multiple organ dysfunction. PMID- 12781865 TI - Ethanol locking as a possible treatment for microbial contamination of long-term central venous catheters. PMID- 12781866 TI - Tumour suppressors hamartin and tuberin: intracellular signalling. AB - Tumour suppressors hamartin and tuberin, encoded by tuberous sclerosis complex 1(TSC1) and TSC2 genes, respectively, are critical regulators of cell growth and proliferation. Mutations in TSC1 and TSC2 genes are the cause of an autosomal dominant disorder known as tuberous sclerosis complex (TSC). Another genetic disorder, lymphangioleiomyomatosis (LAM), is also associated with mutations in the TSC2 gene. Hamartin and tuberin control cell growth by negatively regulating S6 kinase 1 (S6K1) and eukaryotic initiation factor 4E binding protein 1 (4E BP1), potentially through their upstream modulator mammalian target of rapamycin (mTOR). Growth factors and insulin promote Akt/PKB-dependent phosphorylation of tuberin, which in turn, releases S6K1 from negative regulation by tuberin and results in the activation of S6K1. Although much has been written regarding the molecular genetics of TSC and LAM, which is associated with either the loss of or mutation in the TSC1 and TSC2 genes, few reviews have addressed the intracellular signalling pathways regulated by hamartin and tuberin. The current review will fill the gap in our understanding of their role in cellular signalling networks, and by improving this understanding, an integrated picture regarding the normal function of tuberin and hamartin is beginning to emerge. PMID- 12781867 TI - Phosphorylation of initiation factor 4E is resistant to SB203580 in cells expressing a drug-resistant mutant of stress-activated protein kinase 2a/p38. AB - Previous work has shown that increased phosphorylation of eukaryotic initiation factor (eIF) 4E at Ser209 in the C-terminal loop of the protein is observed in response to cellular stress. SB203580, a cell permeable inhibitor of stress activated protein kinase 2a (SAPK2a/p38), suppresses this response in a number of cell types. To validate the in vivo specificity of this inhibitor for the investigation of signalling pathways, which modulate the phosphorylation of eIF4E, we have used 293 cells which inducibly express either a wild-type form (WT SAPK2a) or a drug-resistant mutant of SAPK2a (DR-SAPK2a). These data show that while the arsenite-induced increase in the phosphorylation of eIF4E and hsp25 was sensitive to SB203580 in cells expressing WT-SAPK2a, these responses to SB203580 were abrogated in cells expressing DR-SAPK2a. In addition, the phosphorylation of the eIF4E kinase, MAP kinase integrating kinase-1 (Mnk1), which is activated in response to growth factors or stress, was insensitive to SB203580 in DR-SAPK2a expressing cells. However, a cell-permeable, specific inhibitor of Mnk1, CGP57380 and the phosphatidylinositol-3-kinase (PI3-K) inhibitor, LY294002, prevented eIF4E phosphorylation in 293 cells irrespective of SAPK2a expression. Therefore, this study validates the use of SB203580 for investigating signalling pathways modulating the phosphorylation of eIF4E in cultured cells. PMID- 12781868 TI - T cell receptor-independent CD4 signalling: CD4-MHC class II interactions regulate intracellular calcium and cyclic AMP. AB - CD4 is a coreceptor on T helper (Th) cells that interacts with MHC class II molecules (MHCII). The mechanisms mediating the effects of CD4 on responses by T helper cells to stimulation of the antigen-specific T cell receptor (TCR) are still poorly understood. Here, we demonstrate T cell costimulation via CD4 signalling independent of T cell receptor-mediated signals. Incubation of T helper cells with peptide mimetics of the CD4-binding region on the MHC class II beta2 domain caused intracellular calcium mobilization in the absence of antigen or other T cell receptor stimuli. Engagement of CD4 by peptide mimetics or wild type MHC class II, but not by mutant MHC class II molecules incapable of engaging CD4, inhibited the T cell receptor-mediated increase in cyclic AMP (cAMP) concentrations in T helper cells. CD4-mediated signals activated cyclic AMP phosphodiesterases (PDEs) and inhibited adenylyl cyclase. Full activation and clonal expansion of antigen-stimulated T helper cells required the CD4-mediated regulation of cyclic AMP. Our results suggest a costimulatory mechanism of CD4 function that acts on the second messengers, calcium and cyclic AMP. PMID- 12781870 TI - Involvement of proteinase-activated receptor-2 in mast cell tryptase-induced barrier dysfunction in bovine aortic endothelial cells. AB - We report here a direct modulation by mast cell tryptase of endothelial barrier function through activation of proteinase-activated receptor-2 (PAR-2). In cultured bovine aortic endothelial cells (BAECs), tryptase, trypsin and PAR-2 activating peptide impaired the barrier function as determined by the permeability of protein-conjugated Evans blue. The tryptase-induced barrier dysfunction was completely blocked by U73122, and partially reversed by xestospongin C, calphostin C or Y27632. The intracellular Ca(2+) was elevated by tryptase. It was notable that ioxaglate, a contrast material that degranulates mast cells, markedly increased the permeability when applied to BAECs in combination with mast cells, an action that was blocked by nafamostat, a potent tryptase inhibitor. Immunofluorescence analysis showed that actin stress fibre formation and disruption of VE-cadherin were observed after exposure to tryptase or ioxaglate in combination with mast cells. Therefore, it is suggested that mast cell tryptase impairs endothelial barrier function through activation of endothelial PAR-2 in a manner dependent on the phospholipase C activity. PMID- 12781869 TI - Dissociation of cGMP accumulation and relaxation in myometrial smooth muscle: effects of S-nitroso-N-acetylpenicillamine and 3-morpholinosyndonimine. AB - In guinea pig, primate and man, nitric oxide (NO)-induced regulation of myometrial smooth muscle contraction is distinct from other smooth muscles because cyclic guanosine 3',5'-cyclic monophosphate (cGMP) accumulation is neither necessary nor sufficient to relax the tissue. To further our understanding of the mechanism of action of NO in myometrium, we employed the NO donors, S-nitroso-N-acetylpenicillamine (SNAP), and 3-morpholinosyndonimine (SIN 1) proposed to relax airway smooth muscle by disparate mechanisms involving elevation in intracellular calcium ([Ca(2+)](i)) or cGMP accumulation, respectively. Treatment of guinea pig myometrial smooth muscle with either NO donor at concentrations thought to produce maximal relaxation of smooth muscles resulted in significant elevations in cGMP that were accompanied by phosphorylation of the cGMP-dependent protein kinase substrate vasodilator stimulated phosphoprotein (VASP), shown here for the first time to be present and phosphorylated in myometrium. Stimulation of myometrial strips with oxytocin (OT, 1 microM) produced an immediate increase in contractile force that persisted in the continued presence of the agonist. Addition of SNAP (100 microM) in the presence of OT relaxed the tissue completely as might be expected of an NO donor. SIN-1 failed to relax the myometrium at any concentration tested up to 300 microM. In Fura-2 loaded myometrial cells prepared from guinea pig, addition of SNAP (100 microM) in the absence of other agonists caused a significant, reproducible elevation of intracellular calcium while SIN-1 employed under the same conditions did not. Our data further support the notion that NO action in myometrium is distinct from that in other smooth muscles and underscores the possibility that discrete regional changes in [Ca(2+)](i), rather than cGMP, signal NO-induced relaxation of the muscle. PMID- 12781871 TI - Analysis of subcellular calcium signals in T-lymphocytes. AB - Subcellular Ca(2+) signals were analysed in Jurkat and peripheral human T lymphocytes by confocal Ca(2+) imaging employing an off-line deconvolution method. Stimulation of the TCR/CD3 complex in T-lymphocytes resulted in a series of subcellular pacemaker Ca(2+) signals preceding the first global Ca(2+) signal. The pacemaker signals occurred in a cytosolic "trigger" zone, which is localised close to the plasma membrane. The pacemaker signals were almost independent of extracellular Ca(2+) as shown by measurements in the absence of extracellular Ca(2+), or in the presence of the Ca(2+) channel blocker SK-F 96365. Analysis of the confocal Ca(2+) images revealed characteristic amplitudes of 82 +/- 30 to 109 +/- 21 nM, signal diameters between 2.5 +/- 0.9 and 3.5 +/- 1.5 microm and frequencies between 0.235 and 0.677 s(-1). Taken together, our data constitute the first analysis of subcellular Ca(2+) signals in T cells and indicate that the pacemaker Ca(2+) release events, which are necessary for the development of the global Ca(2+) signal, are composed of Ca(2+) release both from inositol 1,4,5 trisphosphate- and ryanodine receptors. PMID- 12781872 TI - Expression of alternatively spliced interleukin-1 receptor accessory protein mRNAs is differentially regulated during inflammation and apoptosis. AB - Two alternative splice variants of the interleukin-1 receptor accessory protein (IL-1RAcP) mRNA are known. Membrane-bound IL-1RAcP (mIL-1RAcP) promotes intracellular interleukin-1 (IL-1) signalling whereas soluble IL-1RAcP (sIL 1RAcP) is probably an inhibitor of IL-1 signalling. Here we establish that sIL 1RAcP mRNA levels increase 16-fold in response to phorbol esters in the human hepatoma cell line HepG2 via a mechanism that depends on de novo protein synthesis. Following exposure of cells to UV light, a potent inducer of apoptosis, mIL-1RAcP mRNA is rapidly down-regulated and a new steady-state level established briefly before a gradual return to pretreatment levels. Following treatment with staurosporine, also an inducer of apoptosis, mIL-1RAcP mRNA levels steadily decrease through 72 h, with little change in sIL-1RAcP mRNA levels. A novel alternative splice variant, sIL-1RAcP-beta, was identified. Its sequence indicates that sIL-1RAcP-beta is secreted and has a unique second half of the third immunoglobulin (Ig) domain. The dramatic changes in levels of IL-1RAcP mRNAs suggest important functions in regulating sensitivity to IL-1 during stress and may play a role in oncogenic processes that are engaged during chronic inflammation. PMID- 12781874 TI - Honor, respect, and passion. PMID- 12781875 TI - The surgical error examination is a novel method for objective technical knowledge assessment. AB - BACKGROUND: Objective analysis of surgical skill is necessary. A novel method of assessment using simple error analysis in synthetic models is examined for construct validity. METHODS: Two examination protocols were devised using synthetic models. These contained either a purpose made error or were representative of good surgical practice. Protocol one contained models of skin closure and minor operations. Protocol two in addition more complex procedures. Face validity was established by the approval of senior surgeons. Junior surgeons were recruited to undertake the assessment. A p value of less than 0.05 was deemed to be significant. RESULTS: Eighty-nine surgeons were recruited. Both protocol one and two were able to discriminate between groups at statistically significant levels. CONCLUSIONS: Construct validity has been established by showing that error analysis is able to distinguish surgeons with varying levels of experience. PMID- 12781873 TI - Functional interactions between native Gs-coupled 5-HT receptors in HEK-293 cells and the heterologously expressed serotonin transporter. AB - In HEK-293 cells, serotonin (5-hydroxytryptamine, 5-HT) was found to induce cAMP production showing pharmacological characteristics consistent with the 5-HT(7) receptor. The presence of 5-HT(7) (and 5-HT(6)) receptor mRNA was confirmed by RT PCR. Stable HEK-293 cell lines expressing either wild-type or haemagglutinin (HA) tagged human 5-HT transporter (SERT) were selected and SERT function was confirmed using [3H]5-HT transport. The presence of SERT caused a 10-fold reduction in the potency of 5-HT-induced cAMP production compared to control cells. Downstream signalling by 5-HT(6/7) receptors could be detected as 5-HT induced protein kinase A activation and phosphorylation of MAP kinase and CREB using phospho-specific antibodies. SERT inhibitors reversed the reduction in potency of 5-HT-induced cAMP production caused by the presence of SERT, resulting in a concentration-dependent left shift in EC(50) values but also a progressive decrease in the maximal response. Thus, when antidepressants were used to block SERT activity, 5-HT receptor signalling was effectively clamped within a mid range. PMID- 12781876 TI - Long tube decompression is successful in 90% of patients with adhesive small bowel obstruction. AB - BACKGROUND: For patients with small bowel obstruction (SBO), who do not have strangulation obstruction or other contraindications, long tube decompression has been successful in 75% in two studies. In a 1995 prospective randomized study, comparing nasogastric suction (short tube) with long tube decompression, the short tube was successful in 51% and the long tube was successful in 75%. Using upper gastrointestinal endoscopy, a long tube can be advanced into the jejunum in 20 minutes, so the delay in function has been eliminated. METHODS: There were 35 patients with 37 episodes of SBO. From 1983 to 1988, three tubes then available were advanced endoscopically into the jejunum in 17 patients. From 1989 to 2002, an improved tube designed for endoscopic placement was used in 20 patients. RESULTS: From 1983 to 1988 using three tubes, long tube decompression was successful in 12 of 17 (70%); from 1989 to 2002 with the improved tube, decompression was successful in 18 of 20 (90%). CONCLUSIONS: For patients with SBO due to adhesions, a trial with long tube decompression for 48 to 72 hours is recommended. For those who fail a trial with the long tube, laparotomy with enterolysis or bowel resection is indicated. If the operative findings indicate a high risk for recurrent obstruction, then long tube splinting of the small bowel should be considered. PMID- 12781877 TI - Videotape review leads to rapid and sustained learning. AB - BACKGROUND: Performance review using videotapes is a strategy employed to improve future performance. We postulated that videotape review of trauma resuscitations would improve compliance with a treatment algorithm. METHODS: Trauma resuscitations were taped and reviewed during a 6-month period. For 3 months, team members were given verbal feedback regarding performance. During the next 3 months, new teams attended videotape reviews of their performance. Data on targeted behaviors were compared between the two groups. RESULTS: Behavior did not change after 3 months of verbal feedback; however, behavior improved after 1 month of videotape feedback (P <0.05) and total time to disposition was reduced by 50% (P <0.01). This response was sustained for the remainder of the study. CONCLUSIONS: Videotape review can be an important learning tool as it was more effective than verbal feedback in achieving behavioral changes and algorithm compliance. Videotape review can be an important quality assurance adjunct, as improved algorithm compliance should be associated with improved patient care. PMID- 12781878 TI - Resident self-assessment of operative performance. AB - BACKGROUND: In medicine, the development of expertise requires the recognition of one's capabilities and limitations. This study aimed to verify the accuracy of self-assessment for the performance of a surgical task, and to determine whether self-assessment may be improved through self-observation or exposure to relevant standards of performance. METHODS: Twenty-six senior surgical residents were videotaped performing a laparoscopic Nissen fundoplication in a pig. Experts rated the videos using two scoring systems. Subjects evaluated their performances after performance of the Nissen, after self-observation of their videotaped performance, and after review of four videotaped "benchmark" performances. RESULTS: Expert interrater reliability was 0.66 (intraclass correlation coefficient). The correlation between experts' and residents' self-evaluations was initially moderate (r = 0.50, P <0.01), increasing significantly after the residents reviewed their own videotaped performance to r = 0.63 (Deltar = 0.13, P <0.01), yet did not change after review of the benchmarks. CONCLUSIONS: Self observation of videotaped performance improved the residents' ability to self evaluate. PMID- 12781879 TI - Local recurrence in patients with large and locally advanced breast cancer treated with primary chemotherapy. AB - BACKGROUND: Primary chemotherapy is being given in the treatment of large and locally advanced breast cancers, but a major concern is local relapse after therapy. This paper has examined patients treated with primary chemotherapy and surgery (either breast-conserving surgery or mastectomy) and has examined the role of factors which may indicate those patients who are subsequently more likely to experience local recurrence of disease. METHODS: A consecutive series of 173 women, with data available for 166 of these, presenting with large and locally advanced breast cancer (T2>/=4 cm, T3, T4, or N2) were treated with primary chemotherapy comprising cyclophosphamide, vincristine, doxorubicin, and prednisolone and then surgery (either conservation or mastectomy with axillary surgery) followed by radiotherapy were examined. RESULTS: The clinical response rate of these patients was 75% (21% complete and 54% partial), with a complete pathological response rate of 15%. A total of 10 patients (6%) experienced local disease relapse, and the median time to relapse was 14 months (ranging from 3 to 40). The median survival in this group was 27 months (ranging from 13 to 78). In patients having breast conservation surgery, local recurrence occurred in 2%, and in those undergoing mastectomy 7% experience local relapse of disease. Factors predicting patients most likely to experience local recurrence were poor clinical response and residual axillary nodal disease after chemotherapy. CONCLUSIONS: Excellent local control of disease can be achieved in patients with large and locally advanced breast cancers using a combination of primary chemotherapy, surgery and radiotherapy. However, the presence of residual tumor in the axillary lymph nodes after chemotherapy is a predictor of local recurrence and patients with a better clinical response were also less likely to experience local disease recurrence. The size and degree of pathological response did not predict patients most likely to experience recurrence of disease. PMID- 12781880 TI - Surgery or endoscopy for treatment of postcholecystectomy bile duct strictures? AB - BACKGROUND: Surgery is considered the treatment of choice for postoperative biliary strictures. Recently, endoscopic stent placement has been proposed as an alternative to surgical management in selected patients. METHODS: In this retrospective study, 157 patients with postoperative biliary strictures were included. Eighty patients (group A) were treated endoscopically and 77 by surgery (group B). Baseline characteristics of patients were comparable in both groups. Endoscopic therapy consisted of placement of endoprostheses, with trimonthly elective exchange for a 1-year period. Surgical therapy consisted of constructing a biliary-digestive anastomosis in normal ductal tissue. Data were evaluated according to intention-to treat analysis. RESULTS: Successful treatment was achieved in 54% of group A and 73% of group B (P <0.001). Overall 31% of patients developed complications in group A and 23% of patients in group B (P <0.05). However, the rates of severe complications were comparable in both groups (11% versus 13%; P = not significant) In group A the mortality rate was 0% compared with 8% of group B (P <0.05). Recurrent stenosis was evidenced in 6% of patients of group A and 5% of patients of group B. CONCLUSIONS: Surgery provides a better long-term outcome over the endoscopy, because of patients with total obstruction are not amenable to endoscopic approach. When successfully done, endoscopic results are similar to surgical results with less mortality. PMID- 12781881 TI - A loop technique for the safe, secure, and convenient fixation of subclavian central venous catheters to the chest wall. AB - Central venous catheters are placed frequently at our institution. Residents are taught the technique of subclavian line placement starting in their first year of training. Frequently the teaching stops once the line is in the vein. We have developed a method of fixation for subclavian central venous catheters that provides a safe, secure, and convenient means of fixation to the chest wall. The central venous catheter can be inserted by that technique with which the physician is the most comfortable and familiar. PMID- 12781882 TI - A meta-analysis of randomized controlled trials that compared neoadjuvant chemoradiation and surgery to surgery alone for resectable esophageal cancer. AB - BACKGROUND: Esophagectomy is a standard treatment for resectable esophageal cancer but relatively few patients are cured. Combining neoadjuvant chemoradiation with surgery may improve survival but treatment morbidity is a concern. We performed a meta-analysis of randomized controlled trials (RCTs) that compared the use of neoadjuvant chemoradiation and surgery with the use of surgery alone for esophageal cancer. METHODS: Medline and manual searches were done to identify all published RCTs that compared neoadjuvant chemoradiation and surgery with surgery alone for esophageal cancer. A random-effects model was used and the odds ratio (OR) was the principal measure of effect. Systematic quantitative review was done for outcomes unique to the neoadjuvant chemoradiation treatment group, such as pathological complete response. RESULTS: Nine RCTs that included 1,116 patients were selected with quality scores ranging from 1 to 3 (5-point Jadad scale). Odds ratio (95% confidence interval [CI]; P value), expressed as chemoradiation and surgery versus surgery alone (treatment versus control; values <1 favor chemoradiation-surgery arm), was 0.79 (0.59, 1.06; P = 0.12) for 1-year survival, 0.77 (0.56, 1.05; P = 0.10) for 2-year survival, 0.66 (0.47, 0.92; P = 0.016) for 3-year survival, 2.50 (1.05, 5.96; P = 0.038) for rate of resection, 0.53 (0.33, 0.84; P = 0.007) for rate of complete resection, 1.72 (0.96, 3.07; P = 0.07) for operative mortality, 1.63 (0.99, 2.68; P = 0.053) for all treatment mortality, 0.38 (0.23, 0.63; P = 0.0002) for local regional cancer recurrence, 0.88 (0.55, 1.41; P = 0.60) for distant cancer recurrence, and 0.47 (0.16, 1.45; P = 0.19) for all cancer recurrence. A complete pathological response to chemoradiation occurred in 21% of patients. The 3-year survival benefit was most pronounced when chemotherapy and radiotherapy were given concurrently (OR 0.45, 95% CI 0.26 to 0.79, P = 0.005) instead of sequentially (OR 0.82, 95% CI 0.54 to 1.25, P = 0.36). CONCLUSIONS: Compared with surgery alone, neoadjuvant chemoradiation and surgery improved 3-year survival and reduced local-regional cancer recurrence. It was associated with a lower rate of esophageal resection, but a higher rate of complete (R0) resection. There was a nonsignificant trend toward increased treatment mortality with neoadjuvant chemoradiation. Concurrent administration of neoadjuvant chemotherapy and radiotherapy was superior to sequential chemoradiation treatment scheduling. PMID- 12781883 TI - International prospective evaluation of scintimammography with (99m)technetium sestamibi. AB - BACKGROUND: The purpose of this study is to evaluate the efficacy of scintimammography with (99m)Technetium-Sestamibi for the diagnosis of breast cancer. METHODS: This was a multicenter prospective cohort clinical trial. A total of 1,734 women were enrolled of whom 1,243 had complete data upon study completion. RESULTS: The mean +/- standard error age of the patients is 56 +/-12 years (with a range of 19 to 94). Mammographic results were classified by the Breast Imaging Reporting and Data System (BIRADS) as 199 (16%) BIRADS 5, 149 (12%) BIRADS 4, 199 (16%) BIRADS 3, and 696 (56%) BIRADS 2 or 1. Scintimammography was positive for 322 (26%) of the patients and negative for 921 (76%). Histopathology showed malignancy for 201 (16%) of the patients. Sensitivity and specificity of scintimammography was estimated 93% and 87% respectively. A positive predictive value (PPV) of 58% with a negative predictive value of 98% were calculated. CONCLUSIONS: The present study suggests that scintimammography with (99m)Technetium-Sestamibi is highly accurate for the detection of breast cancer. PMID- 12781884 TI - Interleukin-18 concentration in the peritoneal fluid correlates with the severity of peritonitis. AB - BACKGROUND: Interleukin (IL)-18 is a novel cytokine that has recently been characterized as an inducer of interferon-gamma (IFN-gamma). The aim of this study was to investigate the clinical significance of peritoneal IL-18 concentrations in patients with peritonitis. METHODS: We measured IL-18, IFN gamma, and IL-10 concentrations in the peritoneal fluid of 28 patients undergoing laparotomy for peritonitis. Correlations between the peritoneal cytokine concentrations and the severity of illness determined by systemic inflammatory response syndrome (SIRS) criteria, Acute Physiological and Chronic Health Evaluation II (APACHE II) score, peritoneal fluid bacterial culture results, subsequent development of organ failure, and length of hospital stay were assessed. RESULTS: Interleukin-18 concentration was significantly increased in patients who developed SIRS, in those with culture-positive peritonitis, and in those who developed organ failure, as compared with the other patients. Interleukin-10 concentration, which was also significantly increased in patients with culture-positive peritonitis, showed a close correlation with IL-18 concentration. Although a weak correlation was observed between IL-18 and IFN gamma concentrations, IFN-gamma concentrations did not show any association with patients' clinical parameters. However, the IFN-gamma/IL-18 ratio was significantly lower in patients with an APACHE II >/=10, and in those with culture-positive peritonitis, as compared with the other patients. BACKGROUND: Peritoneal IL-18 concentration increased in response to intraperitoneal bacterial infection and seemed to reflect the severity of peritonitis. However, the capacity of IL-18 to produce IFN-gamma may be altered depending on the severity of peritonitis. PMID- 12781885 TI - A prospective comparison between multidetector-row computed tomography and magnetic resonance imaging in the preoperative evaluation of rectal carcinoma. AB - BACKGROUND: Multidetector-row computed tomography (MDCT, or multislice CT) is a new modality with four detectors, which makes examination time shorter and produces higher resolution and multiplanar reformation of the images. Its diagnostic role in patients with rectal carcinoma has not been determined. METHODS: Twenty-one patients with rectal carcinoma were preoperatively examined by both MDCT and magnetic resonance imaging (MRI). Diagnostic accuracies of both modalities were compared regarding depth of tumor invasion and lymph node metastasis based on the pathologic findings. RESULTS: Both examinations detected all tumors. Regarding depth of tumor invasion, the concordance was 95.2% (20 of 21) for MDCT and 100% (21 of 21) for MRI. Regarding lymph node metastasis, the overall accuracy was 61.9% for MDCT and 70.0% for MRI. CONCLUSIONS: Multidetector row computed tomography was equal to MRI in the preoperative local staging of rectal carcinoma. PMID- 12781886 TI - Gastroesophageal reflux, quality of life, and satisfaction in patients with achalasia treated with open cardiomyotomy and partial fundoplication. AB - BACKGROUND: Cardiomyotomy, often associated with an antireflux technique, is effective in the management of achalasia, although gastroesophageal reflux (GER) may occur after the procedure. Patient-centered measures, ie, health-related quality of life (HRQoL) and satisfaction, should be included in the evaluation of the patients. METHODS: A study was made of the incidence of GER (symptoms, upper endoscopy and 24-hour pH monitoring), HRQoL (Short Form-36 Health Survey), and satisfaction after open-access cardiomyotomy and 180-degree anterior fundoplication in 28 consecutive patients, with a minimum postoperative follow-up of 12 months. RESULTS: Mean age was 45 years (range 15 to 80) and 68% were female. In 8 subjects (all with heartburn) GER morbidity was present (4 with esophagitis and 4 with positive pH study), and 6 patients required proton pump inhibitors. Short Form-36 scores after surgery were similar to those found in the general population. Patient satisfaction was high and was more related to the absence of dysphagia than to the presence of GER symptoms. CONCLUSIONS: Gastroesophageal reflux is relatively frequent after cardiomyotomy and partial fundoplication, although the efficacy of proton pump inhibitor treatment minimizes its clinical significance. PMID- 12781887 TI - Successful embolization of a bleeding pseudoaneurysm of the celiac tripod. PMID- 12781888 TI - Indication for peritoneal biopsy in tuberculous peritonitis. AB - BACKGROUND: With the introduction of effective antituberculous chemotherapy, the clinical outcome of tuberculous peritonitis depends much on the diagnostic accuracy of this disease entity. This review summarizes the current state-of-the art thinking regarding the protean manifestation and diagnostic modalities of this major infectious disease. DATA SOURCES: This review was compiled after an extensive search of the current and historical literature, comprising 1,070 cases of tuberculous peritonitis. A number of important areas were highlighted, with emphasis on the diagnostic value and clinical impact of peritoneal biopsy. CONCLUSIONS: We believe an aggressive diagnostic approach, particularly with peritoneal biopsy, is warranted for the diagnosis and timely treatment of tuberculous peritonitis. PMID- 12781889 TI - A prospective controlled double-blind trial of great auricular nerve preservation at parotidectomy. AB - BACKGROUND: Earlier reports of the advantages of preservating the posterior branches of the great auricular nerve (GAN) at parotidectomy were conflicting. This prospective study was aimed at clarifying the controversy. METHODS: Eighty one patients in a university otolaryngology department were recruited. The posterior branches were preserved whenever initial dissection showed that tumor clearance would not be compromised. Touch-pressure sensation was monitored in predefined territories supplied by the GAN using a Semmes-Weinstein aesthesiometer, preoperatively and postoperatively. Minimal pressure thresholds obtained were compared between the two groups. RESULTS: Preservation of the GAN was achievable in 69% of patients; sensory deficit was transient. With the GAN divided, measurable sensory depression occurred up to 2 years after surgery. The difference is statistically significant. Patients' subjective assessment of numbness also conformed to these trends. Additional time taken for preservation of the GAN was about 10 minutes. CONCLUSIONS: The posterior branches should always be preserved if tumor clearance is not compromised. PMID- 12781890 TI - Penetrating injuries of the subclavian artery. AB - BACKGROUND: Penetrating injuries of the subclavian artery occurs infrequently but represent a surgical challenge. We reviewed our experience with penetrating injury of the subclavian artery and identify factors that influenced morbidity and mortality. METHODS: A retrospective review was performed on 54 consecutive patients who sustained penetrating injury to the subclavian artery during a 10 year period. RESULTS: The causes of injuries were gunshot wounds in 46 patients (85%), stab wounds in 5 patients (9%), and shotgun wounds in 3 patients (6%). The overall mortality was 39%. Operative management of the subclavian artery injury included primary repair in 38 patients, interposition grafting in 13 patients, and ligation in 3 patients. The most common associated injury was subclavian vein (44%) followed by brachial plexus (31%). Predictors of survivability include mechanism of penetrating injuries, hemodynamic status of patients on arrival, and three or more associated injuries involving other structures. Associated brachial plexus injury accounts for the majority of long-term morbidity in survivors. CONCLUSIONS: Penetrating injuries of the subclavian artery are associated with high morbidity and mortality. Multiple concomitant injuries, unstable vital signs upon presentation, and gun shot injuries greatly increase mortality. PMID- 12781891 TI - Technical considerations for transabdominal loading of the circular stapler in laparoscopic Roux-en-Y gastric bypass. AB - BACKGROUND: Laparoscopic Roux-en-Y gastric bypass is being undertaken with increasing frequency. We describe a technique for introducing the anvil of the circular stapler using a totally transabdominal approach. METHODS: One hundred consecutive patients underwent laparoscopic Roux-en-Y gastric bypass in a university-affiliated teaching hospital. RESULTS: The cardiojejunostomy was constructed in all 100 patients using the circular stapler with no complications. No anastomotic leaks were detected postoperatively. CONCLUSIONS: The totally transabdominal approach for introducing the anvil of the circular stapler into the gastric pouch is safe and feasible. PMID- 12781892 TI - Differential nitric oxide synthase expression during hepatic ischemia reperfusion. AB - BACKGROUND: In recent years the important role of nitric oxide in hepatic ischemia-reperfusion injury has been increasingly recognised. The prevailing consensus is that reperfusion injury may be partly the result of decreased production of nitric oxide from endothelial nitric oxide synthase and excessive production of nitric oxide from the inducible isoform. We therefore undertook this study to characterize the expression of different nitric oxide synthase isoforms during hepatic reperfusion. METHODS: Male Wistar rats (n = 6) were subjected to 45 minutes of partial hepatic ischemia (left lateral and median lobes) followed by 6 hours of reperfusion. Control animals (n = 6) were subjected to sham laparotomy. The expression of endothelial and inducible nitric oxide synthase was examined using immunohistochemistry and Western blotting. Liver sections were also stained with nitrotyrosine antibody, a specific marker of protein damage induced by peroxynitrite (a highly reactive free radical formed from nitric oxide). RESULTS: Liver sections from all the control animals showed normal expression of the endothelial isoform and no expression of inducible nitric oxide synthase. Livers from all the animals subjected to hepatic ischemia showed decreased expression of endothelial nitric oxide synthase, and all but one animal from this group showed expression of the inducible isoform both in inflammatory cells and in hepatocytes. Western blotting confirmed these findings. Staining with the antinitrotyrosine antibody was also confined to five liver sections from animals subjected to hepatic ischemia. CONCLUSIONS: During the reperfusion period after hepatic ischemia, endothelial nitric oxide synthase is downregulated while inducible nitric oxide synthase is expressed in both hepatocytes and inflammatory cells. The presence of nitrotyrosine in livers subjected to hepatic ischemia-reperfusion suggests that the expression of inducible nitric oxide synthase plays an important role in mediating reperfusion injury in this model. PMID- 12781893 TI - Abdominal wall endometriomas. AB - BACKGROUND: The diagnosis of abdominal wall endometriomas is often confused with other surgical conditions. METHODS: A retrospective study was made of 12 patients presenting with an abdominal wall mass, which proved to be endometrioma. RESULTS: Of a total of 297 patients of endometriosis treated in our hospital over a 7-year period, 12 (4%) had isolated abdominal wall endometriomas. Their mean age was 29.4 years. The presenting symptoms were abdominal mass (n = 12), cyclical (n = 5) or noncyclic pain (n = 7), dyspareunia and dysmenorrhea (n = 1). All patients had a history of gynecologic operations and presented, after an average of 1.9 years, with a tender mass (average 4 cm) at the previous incision site. Preoperative diagnosis was correct in 4 patients (33%) who presented with a cyclically painful abdominal mass. The others were diagnosed as incisional hernia (n = 4), "abdominal wall tumor" (n = 2), and inguinal hernia (n = 2). All patients underwent wide excision of their endometrioma; 2 required polytetrafluoroethylene patch grafting for the resulting fascial defect. The diagnosis was confirmed at frozen section or conventional histological examination in all patients. At follow-up, ranging from 4 months to 3 years, there was no recurrence of endometrioma. CONCLUSIONS: Scar endometrioma commonly presents as an abdominal mass with noncyclical symptoms. Imaging techniques are nonspecific and needle biopsy may confirm the diagnosis. Wide excision is the treatment of choice for abdominal wall endometrioma as well as for recurrent lesions. PMID- 12781894 TI - H-type congenital tracheoesophageal fistula. PMID- 12781895 TI - Why perform randomized clinical trials for sentinel node surgery for breast cancer? PMID- 12781897 TI - A new technique for performing hepatic resections utilizing a specially designed needle. PMID- 12781899 TI - Early postoperative enteral feeding increases anastomotic strength in a peritonitis model. PMID- 12781900 TI - Novel applications of endo GIA linear staplers during pancreaticoduodenectomy and total pancreatectomy. PMID- 12781902 TI - Trials on blood pressure-lowering and secondary stroke prevention. AB - The risk of stroke is strongly and persistently related to the usual level of both systolic blood pressure (SBP) and diastolic blood pressure (DBP). This relation holds for primary and secondary stroke, both ischemic and hemorrhagic. The Perindopril Protection Against Recurrent Stroke Study (PROGRESS) has now provided definitive evidence that lowering the blood pressure of patients with preexisting cerebrovascular disease (prior stroke or transient ischemic attack [TIA]) also reduces the incidence of secondary stroke. PROGRESS showed that a flexible blood pressure-lowering regimen involving an angiotensin-converting enzyme inhibitor (perindopril) and a diuretic (indapamide) reduces the incidence of stroke, major coronary events, and major vascular events by 28%, 26%, and 26%, respectively. These benefits were associated with an average reduction of 9.0 mm Hg (SBP) and 4.0 mm Hg (DBP). The 28% reduction in stroke incidence translated into a 24% reduction in ischemic stroke and a 50% reduction in hemorrhagic stroke. Combination therapy with perindopril and indapamide decreased blood pressure more effectively than did perindopril monotherapy (mean reduction of 12.3 mm Hg [SBP] and 5.0 mm Hg [DBP] vs 4.9 mm Hg [SBP] and 2.8 mm Hg [DBP], respectively) and was equally effective in reducing stroke risk in patients with and without hypertension. In conclusion, blood pressure-lowering therapy is now established as the most important measure for primary and secondary stroke prevention. Results of PROGRESS suggest that antihypertensive treatment with a combination of perindopril plus indapamide should now be routinely considered for all patients with previous stroke or TIA. PMID- 12781903 TI - Atrial fibrillation: hypertension as a causative agent, risk factor for complications, and potential therapeutic target. AB - Atrial fibrillation and hypertension are 2 prevalent, and often coexistent, conditions in the North American population. Their incidence increases with advancing age, and they are responsible for considerable morbidity and mortality. Although the relation between the 2 conditions has long been known, the treatment of hypertension is not currently a focus in the clinical management of atrial fibrillation. Hypertension is associated with left ventricular hypertrophy, impaired ventricular filling, left atrial enlargement, and slowing of atrial conduction velocity. These changes in cardiac structure and physiology favor the development of atrial fibrillation, and they increase the risk of thromboembolic complications. Conventional therapy of atrial fibrillation has focused on interventions to control heart rate and rhythm and the prevention of stroke through the use of anticoagulant medications. In patients with atrial fibrillation, aggressive treatment of hypertension may reverse the structural changes in the heart, reduce thromboembolic complications, and retard or prevent the occurrence of atrial fibrillation. Specific pharmacotherapy could potentially play a major role in the primary and secondary prevention of atrial fibrillation and its complications. PMID- 12781904 TI - Debate: angiotensin-converting enzyme inhibitors versus angiotensin II receptor blockers--a gap in evidence-based medicine. AB - In this article, 2 leading physicians debate the strength of outcome data on the efficacy of angiotensin-converting enzyme (ACE) inhibitors versus angiotensin II receptor blockers (ARBs) for reducing the incidence of cardiovascular, cerebrovascular, and renovascular events. Dr. Stephen G. Ball notes that the efficacy of ACE inhibitors for reducing the risk for myocardial infarction independent of their effects on blood pressure is controversial. In the Heart Outcomes Prevention Evaluation (HOPE) study, ramipril treatment in high-risk patients was associated with a 20% reduction in the risk for myocardial infarction; mean reduction in blood pressure was 3 mm Hg for systolic blood pressure and 1 mm Hg for diastolic blood pressure. The HOPE investigators propose that the 20% reduction was much greater than would be expected based on the observed blood pressure reduction. However, a meta-regression analysis of blood pressure reduction in >20 antihypertensive therapy outcome trials found that the reduction in myocardial infarction risk with ramipril observed in HOPE was consistent with the modest blood pressure reduction seen with that agent. Nevertheless, there are convincing data for prevention of myocardial infarction with ACE inhibitors in patients with heart failure, including those with heart failure after myocardial infarction, as well as supportive evidence from studies in patients with diabetes mellitus and concomitant hypertension. On the other hand, Dr. William B. White takes the position that ARBs are well-tolerated antihypertensive agents that specifically antagonize the angiotensin II type 1 (AT(1)) receptor and provide a more complete block of the pathologic effects of angiotensin II-which are mediated via the AT(1) receptor-than ACE inhibitors. The Evaluation of Losartan in the Elderly (ELITE) II study and the Valsartan Heart Failure Trial (ValHeFT) suggest that ARBs reduce the risk for mortality in patients with congestive heart failure. The Losartan Intervention for Endpoint (LIFE) Reduction in Hypertension trial also demonstrated beneficial effects of ARBs in the prevention of stroke events. The Irbesartan in Patients with Diabetes and Microalbuminuria (IRMA) study, the Irbesartan Diabetic Nephropathy Trial (IDNT), and the Reduction of Endpoints in NIDDM with the Angiotensin II Antagonist Losartan (RENAAL) study demonstrated significant reductions in the rate of progression of renal disease in patients receiving ARBs, independent of effects on blood pressure. These data support the use of ARBs, in addition to the standard of care, in hypertensive patients with heart failure who are intolerant of ACE inhibitors, and also provide compelling evidence for their use in patients with hypertension and type 2 diabetes. PMID- 12781905 TI - How cost-effective are new preventive strategies for cardiovascular disease? AB - Costs of providing a particular medical service can be measured, but it is more difficult to assess whether the service provides good value for the money spent. Rigorous trials have demonstrated the health benefits connected with interventions for treatment and prevention of cardiovascular disease (CVD), and in-depth analyses of the costs associated with many of those interventions have been performed. Careful use of terminology clearly differentiating among cost minimization (relative costs of proved equivalent therapeutics), cost effectiveness (lives saved or years of life added), and cost-benefit (total net effect in monetary terms) analyses is warranted. Although trials commonly assess clinical effectiveness as reductions in mortality or CVD-specific outcomes, improvement in quality of life may be equally important and is expressed in quality-adjusted life-years. Comparisons between therapies can be assessed as a cost-effectiveness ratio. Extensive cost-effectiveness studies have been conducted on many important cardiovascular therapies: (1) beta-blockers and diuretics for multiple CVD outcomes, mortality, and prevention of recurrent myocardial infarction (MI); (2) statins for both primary and secondary prevention of CVD; (3) enalapril for prevention and treatment of congestive heart failure; (4) tissue plasminogen activator treatment of acute MI; (5) coronary artery bypass graft for left main, single-, and 2-vessel coronary artery disease, or severe angina; (6) physician counseling for smoking; and (7) radiofrequency ablation therapy for Wolff-Parkinson-White syndrome. Therapies considered economically attractive include (1) secondary prevention with statins in hyperlipidemia, (2) smoking cessation programs, (3) primary prevention in treatment of high blood pressure with diuretics and beta-blockers, (4) primary prevention with regular exercise programs, (5) secondary prevention with cardiac rehabilitation, and (6) postinfarction treatment with beta-blockers and angiotensin-converting enzyme (ACE) inhibitors. A recent cost-minimization analysis has been performed showing aspirin to be a "best buy" therapy for secondary prevention of CVD. The Ongoing Telmisartan Alone and in Combination with Ramipril Global Endpoint Trial (ONTARGET) and Telmisartan Randomized Assessment Study in ACE-I Intolerant Patients with Cardiovascular Disease (TRANSCEND) program provide potential opportunities for both cost-minimization and cost-effectiveness analyses. PMID- 12781906 TI - The ongoing telmisartan alone and in combination with ramipril global endpoint trial program. AB - The renin-angiotensin system evolved to maintain volume homeostasis and blood pressure and to prevent ischemia during acute volume loss. But in the present age, these mechanisms are redundant, and the clinical significance of angiotensin II results from its pathologic effects, which are mediated by the angiotensin II type 1 (AT(1)) receptor. Activation of AT(1) receptors has been linked to pathologic processes that contribute to atherosclerosis and ischemic events, including oxidative stress, inflammatory processes, low-density lipoprotein cholesterol trafficking, and prothrombotic states. The Ongoing Telmisartan Alone and in Combination with Ramipril Global Endpoint Trial (ONTARGET) program will compare the efficacy of the angiotensin II receptor blocker (ARB) telmisartan, the angiotensin-converting enzyme (ACE) inhibitor ramipril, and combination therapy with telmisartan plus ramipril for reducing cardiovascular risk. The ARB telmisartan is distinguished by its long duration of action, which compares favorably with some other ARBs and conventional antihypertensives. Ramipril was shown in the Heart Outcomes Prevention Evaluation (HOPE) study to reduce the risk for myocardial infarction (MI) and other cardiovascular events in patients at high risk for cardiovascular events but without heart failure or a low ejection fraction. The ONTARGET program consists of 2 randomized, double-blind, multicenter international trials: a principal trial, ONTARGET, and a parallel trial, Telmisartan Randomized Assessment Study in ACE-I Intolerant Patients with Cardiovascular Disease (TRANSCEND). The treatment arms for the principal ONTARGET study are telmisartan 80 mg, ramipril 10 mg, and combination therapy with telmisartan 80 mg plus ramipril 10 mg; for the parallel study TRANSCEND, the treatment arms are telmisartan 80 mg and placebo. Both trials will assess cardiovascular outcomes in patients at high risk using the same criteria as that of the HOPE study, with a single exception: the TRANSCEND trial will enroll patients who do not tolerate ACE inhibitor treatment. The primary end points in both ONTARGET and TRANSCEND are death caused by cardiovascular disease, acute MI, stroke, and hospitalization because of congestive heart failure. The secondary end points include newly diagnosed heart failure, revascularization, new-onset type 2 diabetes mellitus, nephropathy, cognitive decrease and dementia, and newly diagnosed atrial fibrillation; these will be used for hypothesis generation. PMID- 12781907 TI - Ginsenoside Rg1 increases ischemia-induced cell proliferation and survival in the dentate gyrus of adult gerbils. AB - The dentate gyrus is one of the few areas of the mammalian brain where new neurons are continuously produced in adulthood. Recent studies demonstrated that dentate neurogenesis increased after transient global ischemia and it is suggested that the increased neurogenesis contributes to the recovery of hippocampal function. In the present study, adult Mongolian gerbils were chronically treated with ginsenoside Rg1 after ischemia, and the proliferation of cells in the dentate gyrus was examined. It was proved that bromodeoxyuridine (BrdU)-labeled cells in the dentate gyrus were significantly enhanced in number following Rg1 treatment after 6 min global ischemia. In addition, the number of surviving BrdU-positive cells 40 days after ischemia also increased markedly in the Rg1 group. This suggests that ginsenoside Rg1 delivered to the brain well after stroke may have therapeutic benefits. PMID- 12781908 TI - Bidirectional modulation of long-term potentiation by novelty-exploration in rat dentate gyrus. AB - Previous studies have shown that exploration of a novel environment reverses hippocampal long-term potentiation (LTP). Here we demonstrate a bidirectional modulation of LTP measured in the dentate gyrus of freely moving rats by means of novelty exploration. A transient form of LTP lasting about 6 h was induced in perforant path-granular cell-synapses by a weak tetanization protocol (three bursts of 15 pulses at f=200 Hz). LTP was reversed when non-restricted exploration of a novel environment started 2 min after LTP induction. In contrast, using the same interval and limiting the exploration duration to 1 min led to LTP-prolongation. Furthermore, LTP-reinforcement was also obtained when a not-restricted exploration started within 2 min before tetanization. The observed interplay of LTP-impairing and -enhancing factors may be relevant for the modulation of memory formation by novelty. PMID- 12781909 TI - Effects of sufentanil on the release and metabolism of dopamine and ascorbic acid and glutamate release in the striatum of freely moving rats. AB - The effects of either intraperitoneally (i.p.) or intrastriatally administered sufentanil on the release and metabolism of dopamine (DA) in the rat striatum were evaluated using in vivo microdialysis. Dialysate concentrations of DA and its acidic metabolites dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) were increased following i.p. administration of either clinical anesthetic (20 microg/kg) or clinical analgesic (1 microg/kg) sufentanil doses. In addition, sufentanil also increased uric acid concentrations. In contrast, dialysate ascorbic acid and glutamate concentrations were unaffected. Intrastriatal infusion of sufentanil (250 nM) induced only a short lasting decrease in dialysate DA. Subcutaneous naloxone (1.0 mg/kg) abolished sufentanil-induced increases in dialysate DA, DOPAC+HVA and uric acid; however, naloxone (0.1 mM) failed to affect these increases when infused intrastriatally. These results demonstrate that sufentanil, at clinical doses, increases striatal DA release and oxidative metabolism of both DA and xanthine acting at extrastriatal sites with a mu-receptor-mediated mechanism. PMID- 12781910 TI - Nerve growth factor serum concentrations in healthy human volunteers: physiological variance and stability. AB - Human nerve growth factor (NGF) serum concentrations were measured in a healthy sample of 126 participants by a modified highly sensitive and specific two-site enzyme immunoassay. The measured NGF concentrations differ considerably from a normal distribution. The median NGF concentration was 19.68 pg/ml with an interquartile range of 11.06-41.74 pg/ml, which means that 50% of the NGF levels are in this range. In our healthy sample, we found no gender differences but a slight age-related decrease of NGF (r=-0.1326, P=0.1560). Moreover intraindividual stability of NGF was examined in ten volunteers, where no significant changes of serum NGF concentrations were detected over 4 weeks. This stability of our repetitive measurements over 4 weeks suggests that this neurotrophin may be an intraindividually solid marker at least in human serum. PMID- 12781911 TI - Differential regulation of Toll-like receptor mRNAs in experimental murine central nervous system infections. AB - Toll-like receptors (TLR) play a key role in the recognition of microbial components. We investigated the differential regulation of TLR mRNA expression in bacterial and viral mouse models of central nervous system infection. Streptococcus pneumoniae meningitis led to an enhanced expression of TLR2, TLR4 and TLR9 mRNA. In Escherichia coli meningitis, TLR2, TLR4 and TLR7 mRNA expression was increased and Herpes simplex encephalitis caused a rise of TLR4 mRNA. In organotypic hippocampal cultures treatment with S. pneumoniae R6 led to increased expression of TLR2 and TLR3 mRNA. Our data provide evidence that regulation of TLR mRNA is not fully specific for the molecular patterns of the infectious pathogen. The TLR mRNA regulation observed probably represents a combination of specific response to the causative pathogen and non-specific activation of the innate immune system. PMID- 12781912 TI - Endomorphin-1 causes synovial hypoaemia in rat knee joints via a capsaicin sensitive neural pathway. AB - In joints, synthetic mu-opioids reduce inflammatory changes such as protein extravasation and associated oedema formation. However, the effect of endogenous opioid peptides on other inflammatory processes such as altered tissue blood flow has not been investigated. The present study examined the peripheral effects of the endogenous mu-opioid ligand endomorphin-1 (EM-1) on rat knee joint blood flow using laser Doppler perfusion imaging. Topical application of EM-1 (10(-16)-10( 9) mol) to exposed rat knee joints resulted in a dose-dependent increase in synovial vascular resistance with a maximum rise of 56% occurring with the 10(-9) mol dose. Destruction of unmyelinated articular afferents by capsaicin treatment completely abolished the hypoaemic effects of EM-1. These findings suggest that EM-1 acts peripherally in knee joints to decrease synovial blood flow, and this hypoaemic response is dependent on the presence of capsaicin-sensitive nerves. PMID- 12781913 TI - Heat shock protein 108 mRNA expression during chicken retina development. AB - In a developmental study on the expression of heat shock protein 108 (HSP108) mRNA in the chicken retina, we found different spatial and temporal expressions of HSP108 mRNA in each retinal layer. While intense HSP108 signals were found in the retina neuroblast layer at embryonic day 5 (E5), the ganglion cell population (GC), inner nuclear layer (IN) and pigment epithelium (PE) showed HSP108 expression at E9. At E14, HSP108 signals were reduced versus the previous stages even though signals were still detected in the GC, the IN, the outer nuclear layer and the PE. HSP108 signals were still detectable at the E21 stage, although each retinal layer showed a much differentiated morphology and diminished signal intensity. These results suggest that HSP108 expression might be developmentally regulated throughout eye organogenesis and that it plays a role in ocular development. PMID- 12781914 TI - Hippocampal synaptophysin immunoreactivity is reduced during natural hypothermia in ground squirrels. AB - Natural hypothermia during hibernation results in physiological and behavioral deficits. These changes may be traced at the level of hippocampal signal transduction. We investigated synaptophysin immunoreactivity (SYN-ir) in the hippocampus after short and long periods of hypothermia and short and long periods of euthermy in hibernating ground squirrels. SYN-ir in the stratum lucidum of the hippocampus was transiently reduced during natural hypothermia. Natural hypothermia thus reduces synaptic efficacy. This may play a role in the reduced neuronal connectivity of CA3 pyramidal cell dendrites observed in hibernating ground squirrels. PMID- 12781915 TI - Bupivacaine and ropivacaine suppress glycine- and glutamate-induced ion currents in acutely dissociated rat hippocampal neurons. AB - Bupivacaine and ropivacaine are local surgical anesthetics with great efficacy in post-operative pain relief and labor analgesia. In the present study, the effects of bupivacaine and ropivacaine on ion currents induced by glycine and glutamate in acutely dissociated hippocampal CA1 neurons of rats were investigated via a nystatin-perforated patch clamping method at a clamped voltage. The magnitude of the glycine-induced ion currents was decreased reversibly and in a time-dependent manner by continuous application of 0.1 mg/ml of either bupivacaine or ropivacaine. The magnitude of the glutamate-induced ion currents was also suppressed time-dependently by continuous application of either bupivacaine or ropivacaine. The inhibitory action of bupivacaine and ropivacaine on currents induced by glycine and glutamate could be one of the mechanisms behind the actions of these anesthetics. PMID- 12781916 TI - Differential effects of morphine and cocaine on locomotor activity and sensitization in mu-opioid receptor knockout mice. AB - The present study was undertaken to investigate the hypothesis that the mu-opioid receptors play a crucial role in locomotor activity and sensitization to cocaine and morphine in wild-type and mu-opioid receptor knockout mice. Our results show that morphine and cocaine increased locomotor activity in wild-type mice, but failed to increase locomotor activity in mu-opioid receptor knockout mice, suggesting a contribution of mu-opioid receptor. Repeated morphine treatment induced sensitization in wild-type mice, but this was not observed in mu-opioid receptor knockout mice. In contrast repeated cocaine treatment produced sensitization in mu-opioid receptor knockout mice, but not in wild-type mice on day 6. However, the sensitization to cocaine was observed in mu-opioid receptor knockout and wild-type mice on day 12. These results suggest that the expression of mu-opioid receptor may contribute to locomotor sensitization induced by morphine, but that mu-opioid receptor does not play an important role in mediating sensitization to cocaine. PMID- 12781917 TI - An event-related potentials study of biological motion perception in humans. AB - In order to clarify the neural dynamics involved in the perception of biological motion, we recorded event-related potentials from 12 subjects. The subjects were shown biological motion or scrambled motion as a control stimulus. In the scrambled motion, each point had the same velocity vector as in the biological motion, but the initial starting positions were randomized. The perception of both biological and scrambled motion elicited negative peaks at around 200 (N200) and 240 ms (N240). Furthermore, both negative peaks were significantly larger in the biological motion condition than in the scrambled motion condition over the right occipitotemporal region. In light of previous human neuroimaging studies, we speculate that component N200 is generated near the extrastriate cortex area and N240 is generated from the superior temporal sulcus region. PMID- 12781918 TI - Development of a high-throughput bioassay to screen melatonin receptor agonists using human melatonin receptor expressing CHO cells. AB - Melatonin receptors belong to the superfamily of G-protein-coupled receptors and appear to couple with Gi type of G protein, which has an inhibitory effect on the adenylate cyclase. Normally, melatonin dose not induce transient elevation of intracellular calcium concentration in CHO cells stably expressing melatonin receptors. Accordingly, the cells are unable to be used for fluorescent imaging plate reader (FLIPR), which is the device used to measure the cellular signal as a calcium elevation. To overcome this issue we tried to transfect chimeric G protein, Gqi5, into CHO cells expressing melatonin receptors. The Gqi5 is a chimeric Gq protein containing the five carboxyl-terminal amino acids from Gi, which interact with Gi-coupled receptor and possess the function of evaluating calcium concentration through the Gq pathway. The transfected cells result in a calcium elevation in a concentration-response manner. The specificity of this assay was similar to that of radioreceptor binding assay. Therefore, this FLIPR assay, using melatonin receptor and Gqi5 expressing CHO cells, is available for clinical bioassay of melatonin and for the screening of specific ligands of melatonin. PMID- 12781919 TI - Quantitative histochemical assessment of oxidative metabolism in the subfornical organ after partial aortic ligature in rats. AB - Circumventricular organs are considered to be involved in hydromineral homeostatic responses. In this study we used quantitative histochemistry of cytochrome oxidase to evaluate the oxidative metabolic activity of the subfornical organ of rats with a partial aortic occlusion. These rats showed a significant increase in water intake from the second day after the ligature, while natriophilia was already significant on the first day. Greater levels of cytochrome oxidase activity were found in subfornical organs of partial aortic ligated rats when compared with control, providing further evidence for the involvement of this circumventricular structure in fluid homeostasis at least in this hyperdipsic, hypernatriophilic, hyperreninemic and hypertensive experimental model. PMID- 12781920 TI - Oxygen seizure latency and peroxynitrite formation in mice lacking neuronal or endothelial nitric oxide synthases. AB - Nitric oxide (NO) from endothelial or neuronal NO synthases (eNOS or nNOS) may contribute both to the cerebrovascular responses to oxygen and potentially to the peroxynitrite-mediated toxic effects of hyperbaric oxygen (HBO(2)) on the central nervous system (CNS O(2) toxicity). In mice lacking eNOS or nNOS (-/-), regional cerebral blood flow (rCBF) and 3-nitrotyrosine (3-NT), a biochemical marker for peroxynitrite (ONOO(-)) formation, were measured in the brain during HBO(2) exposure. These variables were then correlated with EEG spiking activity related to CNS O(2) toxicity. In wild-type (WT) mice, HBO(2) exposure transiently reduced rCBF, but by 60 min rCBF was restored to baseline levels and above, followed by EEG spikes. Mice lacking nNOS also showed initial depression of rCBF followed by hyperemia but the delay in the onset of EEG discharges was greater. In contrast, in eNOS-deficient mice rCBF did not decrease and hyperemia was less pronounced during HBO(2). EEG spike latency was longer in eNOS(-/-) compared to WT or nNOS( /-) mice. 3-NT gradually increased in all strains during HBO(2) but accumulation was slower in nNOS(-/-) mice, consistent with less ONOO(-) production. These results indicate that NOS-deficient mice have different cerebrovascular responses and tolerance to HBO(2) depending on which enzyme isoform is affected. The data suggest a key role for eNOS-dependent NO production in cerebral vasoconstriction and in the development of hyperoxic hyperemia preceding O(2) seizures, whereas neuronal NO may mediate toxic effects of HBO(2) mainly by its reaction with superoxide to generate the stronger oxidant, peroxynitrite. PMID- 12781921 TI - Modification of cardiovascular response of posterior hypothalamic adenosine A(2) receptor stimulation by adenylate cylase, guanylate cyclase and by K(ATP) channel blockade in anesthetized rats. AB - Cardiovascular inhibitory effects induced by posterior hypothalamic adenosine A(2) receptors and their modulation by nitric oxide were suggested by our previous report. In this experiment, we examined the modulation of cardiovascular effects of adenosine A(2) receptor stimulation by adenylate cyclase, guanylate cyclase and ATP-sensitive K(+) channel in the posterior hypothalamus. Posterior hypothalamic injection of drugs was performed in anesthetized, artificially ventilated male Sprague-Dawley rats. Injection of adenosine A(2) receptor agonist 5'-(N-cyclopropyl)-carboxamidoadenosine (CPCA; 1, 2 and 5 nmol) produced a dose dependent decrease of blood pressure and heart rate. Pretreatment with adenosine A(2) receptor antagonist 3,7-dimethyl-1-propargylxanthine (10 nmol) blocked the depressor and bradycardiac effects of CPCA (5 nmol). Pretreatments with adenylate cyclase inhibitor MDL-12330 (10 nmol) and guanylate cyclase inhibitor LY-83583 (5 nmol) attenuated the depressor and bradycardiac effects of CPCA (5 nmol). In addition, pretreatment with ATP-sensitive K(+) channel blocker glipizide (20 nmol) attenuated the depressor and bradycardiac responses of CPCA (5 nmol). These results suggest that posterior hypothalamic adenosine A(2) receptors play an inhibitory role in the central cardiovascular regulation and that both adenylate cyclase and guanylate cyclase mediate the depressor and bradycardiac actions of adenosine A(2) receptors. Also, ATP-sensitive K(+) channel mediates the posterior hypothalamic cardiovascular regulations of adenosine A(2) receptors. PMID- 12781922 TI - A truncated delta opioid receptor, spontaneously produced in human but not rat neuroblastoma cells, interferes with signaling of the full-length receptor. AB - In addition to the established human delta opioid receptor SH-SY5Y neuroblastoma cells produce an atypical, shorter, form of this receptor which is predicted to lack the third intracellular domain. Hence it will be referred to as hdelta(deltaICD3). Notably, in unaltered human brain tissue only the established ('wild type') delta receptor was detected. After transfection of the human wild type delta receptor (hdelta(wt)) into NG 108-15 rodent neuroblastoma-derived cells, HEK 293 human embryonic kidney cells and NIH 3T3 mouse fibroblasts, all these cell types produced hdelta(deltaICD3). Only the human but not the rat delta opioid receptor was processed, arguing for a high sequence selectivity of the cleavage process. Upon agonist stimulation hdelta(deltaICD3) was not able to activate potassium channels (K(ir)3.1/K(ir)3.4) expressed in Xenopus laevis oocytes. However, hdelta(deltaICD3) dose-dependently inhibited the signaling of hdelta(wt) if co-expressed with the latter. Thus, hdelta(deltaICD3) can be produced by many cell types and, once produced, markedly interferes with normal delta receptor signaling. PMID- 12781923 TI - The dorsal portion of the lumbar intervertebral disc is innervated primarily by small peptide-containing dorsal root ganglion neurons in rats. AB - The intervertebral discs are innervated by the dorsal root ganglion (DRG) neurons. In the present study, we applied Fluoro-Gold (FG) to the dorsal portion of the L5-L6 intervertebral disc to label DRG neurons retrogradely, and then examined whether FG-labeled neurons were substance P (SP)-immunoreactive or isolectin B4 (IB4)-binding. Of the FG-labeled neurons, 44.0% were immunoreactive for SP, whereas only 0.6% were reactive for IB4. The rate of SP-immunoreactive neurons was significantly higher than that of IB4-binding neurons (P<0.001), suggesting that under physiological conditions the dorsal portion of the lumbar disc is mainly innervated by peptide-containing neurons. PMID- 12781925 TI - The client as partner in reproductive health consultations. PMID- 12781924 TI - Ethanol inhibits muscarinic receptor-mediated DNA synthesis and signal transduction in human fetal astrocytes. AB - We previously found that ethanol inhibits muscarinic receptor-induced proliferation of rat cortical astrocytes and human astrocytoma cells and suggested this as a possible mechanism involved in its developmental neurotoxicity. We also observed that, though several signal transduction pathways are relevant for carbachol-induced cell proliferation, activation of PKC zeta and p70S6 kinase is selectively inhibited by low concentrations of ethanol. In the present study we used fetal human astrocytes to expand these findings to a direct target of ethanol in humans. Astrocyte cultures, deriving from legally aborted fetuses, were stained for GFAP and shown to be 90-95% pure. Carbachol induced increases in [(3)H]thymidine and BrdU incorporation in synchronized cells. Carbachol-induced DNA synthesis was strongly inhibited by ethanol. Carbachol also induced phosphorylation of (Thr410)PKC zeta, (Ser473)Akt, and (Thr389)p70S6 kinase, and ethanol (50 mM) inhibited phosphorylation of PKC zeta and p70S6 kinase, but not of Akt. These results expand previous findings in rat astrocytes and human astrocytoma cells and suggest that intracellular signal transduction pathways activated by muscarinic receptors may represent a relevant target for the developmental neurotoxicity of ethanol in humans. PMID- 12781926 TI - Increasing patient participation in reproductive health consultations: an evaluation of "Smart Patient" coaching in Indonesia. AB - Paternalistic models of health care, social distance between patients and providers, and cultural norms discourage patients from playing an active role in health consultations. This study tested whether individual coaching can give family planning patients the confidence and communication skills to talk more openly and more vigorously with providers. Educators met with 384 Indonesian women in clinic waiting rooms and coached them on asking questions, expressing concerns, and seeking clarification. An analysis of audiotaped consultations found that patients who received coaching articulated significantly more questions and concerns than others. Coaching narrowed differentials in active communication by patient type, age, and assertiveness, but it widened differentials by patient education and socioeconomic class. The discontinuation rate at 8 months was lower in the intervention than the control condition, but the difference was only marginally significant. PMID- 12781927 TI - Tailored messages for breast and cervical cancer screening of low-income and minority women using medical records data. AB - Barriers to screening and early detection often result in cancers in low-income and minority women diagnosed at stages too advanced for optimal treatment. This randomized controlled trial examined whether a personalized form (PF) letter containing generic cancer information and a personalized tailored (PT) letter containing minimally tailored individualized risk factor information based on medical records data affected breast and cervical cancer screening among 1574 urban low-income and minority women. The personalized form-letter group was significantly more likely to schedule a screening appointment and to have undergone a Pap test and mammography within 1 year after the intervention than were the tailored letter and control groups (P<0.001 for all comparisons). Personalized tailored letters that contain individualized cancer risk factor information may decrease the likelihood of receiving cancer screening among medically underserved low-income and minority women, but personalized form letters that contain generic cancer information may improve these rates in this disadvantaged population. PMID- 12781928 TI - Stimulating active coping in patients with rheumatic diseases: a systematic review of controlled group intervention studies. AB - Teaching patients with rheumatic diseases to cope actively with their problems may increase the social support they receive and, also, the quality of their lives. In this paper, a systematic review of coping interventions for people with rheumatic diseases is described. Fourteen controlled trials were selected. Effects on quality of life have been measured in 13 studies of which 6 found positive effects. Effects on social support have been found in one of four studies investigating this variable. Coping has been measured in three studies with effects found on active coping in one study. There is a need for well designed research on the effects of active coping in people with rheumatic diseases, as this may open new perspectives in patient education. PMID- 12781929 TI - Comparison of activities and attitudes of general practitioners concerning genetic counseling over a 10-year time-span. AB - The aim of this study was to investigate whether the activities and attitudes of general practitioners (GPs) concerning genetic counseling have changed between 1989 and 1999. In 1989 a random sample of 124 GPs in The Netherlands was selected. Of these GPs, 98 were contacted again in 1999 and 71 completed the questionnaire. The study showed an increase in the percentage of GPs who provided genetic counseling when a risk factor for having a child with a congenital disorder was present and known. In both 1989 and 1999, the GPs seldom used a recommended combination of oral and written information, and only data that was available in the databases on the risk indicator 'use of medication' increased over the years. GPs are still supporters of a directive method of counseling, and seem to believe that the main goal of genetic counseling is to prevent hereditary and congenital disorders. Although, between 1989 and 1999 more GPs provided genetic counseling when a risk indicator was present and known or referred to a clinical geneticist, only limited improvement was found in the activities of GPs to attempt to collect these data. PMID- 12781930 TI - Evaluating the reliability and validity of three tools to assess the quality of health information on the Internet. AB - The quality of Internet information needs to be evaluated and several tools exist for this purpose. However, none have demonstrated reliability and validity. This study tested the internal consistency and validity of the information quality tool (IQT), quality scale (QS) and DISCERN using 89 web sites discussing smoking cessation. The inter-rater reliability of the tools was established by exploring the agreement between two independent raters for 22 (25%) of the sites. The IQT and DISCERN possessed satisfactory internal consistency (as measured by Cronbach's alpha). The IQT, QS and DISCERN showed satisfactory inter-rater reliability (as measured by kappa and intraclass correlations). The IQT, QS and DISCERN correlated positively with each other, supporting the convergent validity of the tools. This study provides some evidence for the reliability and validity of the IQT, QS and DISCERN, although this needs testing in further research with different types of Internet information and larger sample sizes. PMID- 12781931 TI - Underlying dimensions of verbal communication between dentists and patients in prosthetic dentistry. AB - The study explores whether the task-focused and socio-emotional dimensions of clinical communication is recognizable in the verbal communication context of prosthetic dentistry, as well as if there are other dimensions of communication in that context. Sixty-one audio recordings were made at three specialist clinics of prosthetic dentistry in Sweden. Sixty-one patients and 15 dentists participated. Sixty-one prosthetic rehabilitation periods were followed. Tooth- or implant-supported fixed prostheses were placed during this time. One visit near the end of each rehabilitation period was audiorecorded. The verbal communication was analyzed with the Roter Interaction Analysis System (RIAS) dental. The categories were subjected to exploratory factor analysis. Patient and dentist verbal behavior could not be defined exclusively as socio-emotional or task-focused but had to be defined in other dimensions; Emotional exchange, information exchange-patient horizon, relation building exchange, information exchange-dentist horizon, and administrative and counseling exchange. Since the 'emotional exchange' factor was found to capture the largest share of the variance in the communicated patterns, the present study suggests that prosthetic care in dentistry is basically a human relationship with strong emotional communicative content. PMID- 12781932 TI - Assessing the information needs of post-myocardial infarction patients: a systematic review. AB - We systematically reviewed studies examining information needs of post-myocardial infarction patients and their families. Electronic databases and bibliographies of relevant papers were searched and experts in the field contacted to find relevant studies. A standardised data form was used to extract data on study populations, instruments and results. Of 14 published studies, 6 used the same instrument. Information about risk factors ranked as the most important category overall, followed by information on cardiac anatomy and physiology, medications, and physical activity. Information about miscellaneous items, diet, psychological factors, and the CCU, although ranked lower, were still rated important. Some variation between settings was evident. Patients preferred physicians over nurses as information givers. Differences were found between patient and nurse ratings of information categories. Differences were found in the self-perceived information needs of patients responding to different instruments, indicating a priming effect. Changes in ranking of information categories between CCU, PCCU and PD are congruent with decreasing levels of patient dependency. The information needs of women and other post-MI subgroups, including the elderly, minority ethnic groups, patients with severe coronary disease, and deprived patients, have not been assessed. Patients have not been involved in the design of information needs assessment instruments. PMID- 12781933 TI - Effects of psychosocial interventions on quality of life in adult cancer patients: meta analysis of 37 published controlled outcome studies. AB - The present meta analysis summarized the results of 37 published, controlled studies that investigated the effectiveness of psychosocial interventions on quality of life (QoL) in adult cancer patients. The overall effect size of psychosocial interventions and the effect of potential moderating variables such as type and duration of intervention, sociodemographic and clinical parameters, characteristics of QoL measurement, and methodological quality of the selected studies were calculated using a meta analysis model suggested by Hunter and Schmidt. The overall effect size was 0.31 (correlation equivalent delta) which corresponds to a standardized mean difference of d=0.65 (N=3120 cancer patients). The most important moderating variable was duration of psychosocial intervention with durations of more than 12 weeks being significantly more effective than interventions of shorter duration. The meta analytical findings support the usefulness of psychosocial interventions for improving QoL in adult cancer patients. PMID- 12781934 TI - A medication self-management program to improve adherence to HIV therapy regimens. AB - This study examined whether a self-management intervention based on feedback of adherence performance and principles of social cognitive theory improves adherence to antiretroviral dosing schedules. Forty-three individuals with HIV/AIDS who were starting or switching to a new protease inhibitor regimen were randomly assigned to be in a medication self-management program or usual care control group. The self-management program included skills development exercises, three monthly visits for medication consultations, and monthly feedback of adherence performance using electronic monitors on medication bottles. Participants also completed a 40-item questionnaire that measured self-efficacy to take medications, on schedule, in a variety of situations. Logistic regression analysis indicated that individuals in the self-management group were significantly more likely to take 80% or more of their doses each week than individuals in the control group (n=29, OR=7.8, 95% CI=2.2-28.1). Self-management training with feedback of adherence performance is a potentially useful model for improving adherence to complex regimens in HIV/AIDS care. PMID- 12781935 TI - Patient and physician factors predict patients' comprehension of health information. AB - While patients frequently do not comprehend health information, little is known about patient and physician factors that influence lack of comprehension. To assess effectiveness of health information exchange, 19 physicians and 145 patients were given post-encounter questionnaires to evaluate the preceding visit. We analyzed differences in beliefs between patients who comprehended health information and patients who did not, and whether physicians' attitudes and self-assessment of their educational abilities influenced this comprehension. Patients with insufficient comprehension were more likely to have schooling below college and cited language as a barrier. Physicians who believed health information delivery to be important had fewer patients with comprehension difficulties, while physicians who assessed themselves as very effective educators had significantly more patients with lack of comprehension, compared with physicians who did not feel as effective. Patients' comprehension of health information was associated not only with patient factors but also with physicians' attitude and self-assessment. PMID- 12781936 TI - The effect of decision aids on the agreement between women's and physicians' decisional conflict about hormone replacement therapy. AB - The aim of this secondary analysis was to compare the effects of a tailored decision aid (DA) with those of a pamphlet on the agreement between women's and physicians' decisional conflict about hormone replacement therapy (HRT). A total of 40 physicians and 184 women provided data. The agreement between women's and physicians' decisional conflict scores was measured using the intraclass correlation coefficient (ICC). The ICC was higher for dyads in the DA group (ICC=0.44; 95% confidence interval (CI)=0.25-0.59) compared to the pamphlet group (ICC=0.28; 95% CI=0.06-0.47). When the average score of decisional conflict of women nested within a physician and of each physician were used, the ICC for the DA group and the pamphlet group was 0.41 (95% CI=-0.04 to 0.72) and 0.06 (95% CI= 0.41 to 0.49), respectively. Compared to pamphlets, DAs appear to improve the agreement between women's and physicians' decisional conflict about HRT. PMID- 12781937 TI - The effect of growth atmosphere on the ability of Listeria monocytogenes to survive exposure to acid, proteolytic enzymes and bile salts. AB - Four isolates of Listeria monocytogenes from food, human and environmental sources were grown separately in broth (pH 6.0 at 8 degrees C) under atmospheres of air, 100% N(2), 40% CO(2):60% N(2) or 100% CO(2). Exponential and stationary phase cells were harvested to determine if growth atmosphere and growth phase influenced this pathogen's ability to survive exposure to an acid environment coupled with proteolytic enzymes, and the activity of bile salts. In general, isolates were more resistant to the acid environment than the bile salts environment and stationary phase cells were significantly more resistant to both environments than exponential phase cells. Irrespective of prior growth atmosphere, none of the isolates when in exponential phase remained detectable following full exposure to the acid environment (110 min at 37 degrees C) or the bile environment (3 h at 37 degrees C). With the exception of one isolate grown under the atmosphere of 40% CO(2):60% N(2), all isolates when in stationary phase were detectable following full exposure to the acid environment but death rates varied significantly. Stationary phase cells of all isolates grown under 40% CO(2):60% N(2) and 100% CO(2) were highly susceptible to the bile salts environment: cells were not detectable after a 2-min exposure whereas stationary phase cells grown under air or 100% N(2) were recovered following full exposure to the bile environment. Survival curves were characterised by a population decline of at least 3 log(10)/ml (from an initial level of 7 log(10) CFU/ml) in the first 15 min; thereafter a constant population number of approximately 4 log(10)/ml was maintained over the remaining exposure period. No survival was observed when stationary phase cells of L. monocytogenes FRRB 2538 grown in air and 100% N(2) were subjected to the acid environment followed by immediate exposure to the bile salts environment. The results showed that growth atmosphere and growth phase could influence survival of this pathogen against conditions that imitate the extremes of the most important nonspecific defence mechanisms against microbial infection: the acid environment of the stomach coupled with the activity of proteolytic enzymes, and the activity of bile salts in the small intestine. PMID- 12781938 TI - Influence of food preservation parameters and associated microbiota on production rate, profile and stability of acylated homoserine lactones from food-derived Enterobacteriaceae. AB - Quorum-dependent regulation is mediated by N-acyl-L-homoserine lactones (AHLs) in several Gram-negative bacteria. The production of AHLs has typically been studied using pure bacteria cultures grown in nutrient-rich media at optimal temperature. AHLs are produced in several chill-stored foods by Gram-negative bacteria participating in spoilage. As part of our investigation of the role of AHLs in food quality, we studied the AHL production in two Enterobacteriaceae isolated from cold-smoked salmon under growth conditions typical of those found in cold smoked salmon. We tested the influence of carbon source (glucose, sucrose, xylose, arabinose, mannose, mannitol and sorbitol), temperature (5 and 25 degrees C), salt concentration (0-7%), pH (6, 7 and 8) and co-existing lactic acid bacteria microflora on the AHL profile and production rate from Serratia proteamaculans strain B5a and Enterobacter agglomerans strain B6a. The two strains produced the same types of AHLs under all conditions tested. The specific AHL concentrations (moles/liter/OD(450)) changed slightly for both strains at the various conditions. S. proteamaculans strain B5a produced approximately 150 nM/OD(450) N-3-oxo-hexanoyl homoserine lactone (OHHL) and E. agglomerans strain B6a produced two major signals, OHHL and N-3-oxo-octanoyl homoserine lactone (OOHL) in a 1:9 ratio with a total concentration of approximately 3000 nM/OD(450). The AHL signal molecules became unstable with increasing pH (>7.5). In cold-smoked salmon, pH is approximately 6 and therefore only a low degree of pH-induced turnover is expected to occur in this product. Overall, our study demonstrates that food-derived Enterobacteriaceae produce AHLs of the same type and in the same magnitude when grown under food-relevant conditions as when grown in laboratory media at high temperature. Also, the AHLs produced in foods will be relatively stable and their regulatory impact lasting during storage. PMID- 12781939 TI - Theoretical and experimental evidence of extinction and coexistence of killer and sensitive strains of yeast grown as a mixed culture in water. AB - Theoretical and experimental evidences of extinction and coexistence during batch interaction of killer and sensitive strains of Saccharomyces cerevisiae grown as a mixed culture in pure water are provided. The experimental results show that in the limited nutrient conditions of growth in pure water, the killer yeast is subject to extinction when the initial concentration ratio of killer to sensitive yeast is 1:100. However, if the initial concentration ratio of killer to sensitive yeast is 1:1, both strains coexist. Substantial damped oscillations are associated with the growth process in the mixed culture. A new theoretical model that was originally developed for recovering the growth of single species in isolation is extended and applied to two species competing over a common ecological niche. The solutions of the model are shown to recover all the qualitative features captured in the experiments. PMID- 12781940 TI - Origin and spoilage potential of the microbiota dominating genus Psychrobacter in sterile rehydrated salt-cured and dried salt-cured cod (Gadus morhua). AB - Salt-cured and dried salt-cured cod rehydrated using sterile water and equipment have a short shelf life at 4 degrees C due to high bacterial counts. The microbiota develops off-odours which partly can be described as musty, causing sensory rejection within 7-10 days of chilled storage. The microbiota composition was studied in a total of 38 samples obtained from 10 different, both commercial and laboratory produced, salt-cured and dried salt-cured cod products. The dominating bacterium, representing at least 90% of the total viable count in all products studied, was identified as belonging to the genus Psychrobacter; a Gram negative, oxidase- and catalase-positive, nonpigmented, halotolerant, psychrotolerant, facultative aerobe and nonmotile bacterium. The morphology of the bacterium resembles coccobacilli and the cells occur most often in pairs. The bacterium was able to hydrolyze lipids, but not proteins. It did not produce H(2)S or TMA and the spoilage in rehydrated salt-cured and dried salt-cured cod is therefor different from what is observed in fresh cod. However, samples inoculated with Psychrobacter immobilis gave the same musty odour as spoiled control samples but earlier in the storage period and of a stronger intensity. In a field experiment, carried out to investigate the origin of the dominating bacterium, it was found that the microbiota in both sterile rehydrated commercially produced and laboratory (aseptically) produced salt-cured cod was dominated by this same bacterium. The bacterium was also isolated from cod skin mucus immediately after capture. The bacterium survived NaCl concentrations up to 25% (w/v) NaCl, stating its ability to survive during the salt-curing process. The dominating bacterium in rehydrated salt-cured and dried salt-cured cod seems to mainly originate from the fresh fish itself and not from contamination during processing. PMID- 12781941 TI - Specific identification of certain probiotic Lactobacillus rhamnosus strains with PCR primers based on phage-related sequences. AB - PCR primers derived from Lactobacillus rhamnosus phage Lc-Nu genome were used to screen the presence of phage-related sequences in Lb. rhamnosus strains. Several primer pairs derived from structural and replication gene regions of phage Lc-Nu amplified PCR products of expected sizes from bacterial strains revealing phage related sequences in 10 of 11 Lb. rhamnosus strains. Strain-specific PCR primers for three probiotic Lb. rhamnosus strains were derived from these phage-related sequences for identification and detection purposes. Specificity of these primers was tested against 11 Lb. rhamnosus strains and over 40 other bacterial strains. PMID- 12781942 TI - Growth and energy generation by Enterococcus faecium FAIR-E 198 during citrate metabolism. AB - Citrate metabolism by Enterococcus faecium FAIR-E 198, isolated from Greek Feta cheese, was studied in various growth media containing citrate either in the presence of glucose, or as the sole carbon source, both under aerobic and anaerobic conditions. In de Man-Rogosa-Sharpe (MRS) broth with increasing citrate concentrations, cometabolism of citrate and glucose took place. Glucose was stoichiometrically converted into lactate, while citrate into acetate. Glucose consumption and biomass yield were enhanced with increasing initial citrate concentrations, even though maximum specific growth rate was not. When citrate was used as the sole carbon source in increasing initial concentrations, the main end product was acetate. Small amounts of lactate, formate, ethanol, and acetoin were also produced. In all cases, no significant differences were observed between aerobic and anaerobic conditions. However, when citrate was used as sole carbon source, formate production was favored in the absence of oxygen. The present work shows that E. faecium is able to utilize citrate in synthetic media, either in the presence of glucose or as the sole carbon source, resulting in energy production and the formation of aroma compounds. PMID- 12781943 TI - Stress response of Listeria monocytogenes isolated from cheese and other foods. AB - The responses to pH and sodium chloride of four strains of Listeria monocytogenes isolated from Portuguese cheese, with a sodium chloride concentration of about 2% (w/v) and a pH value from 5.1 to 6.2, were studied. Two isolates from meat and two clinical isolates related to food-borne listeriosis, in which the implicated food product had about 2-3.5% (w/v) sodium chloride, also were studied. The effect of temperature on pH and sodium chloride sensitivity was also determined. The results show that natural isolates vary in response to these stresses and the data were often at variance with previously published data. Strains varied in sensitivity to low pH and to high sodium chloride concentration but the cheese isolates tended to be more resistant. A lower temperature was associated with a decrease in resistance to low pH and to sodium chloride. All strains showed an acid tolerance response induction when grown at pH 5.5 and although the time required for maximum induction of the response varied between strains, 2 h of acid adaptation, at least, was necessary which is longer than previously reported. Some strains showed an osmotolerance response after incubation in 3.5% (w/v) sodium chloride. Osmoadaptation, in addition to inducing an osmotolerance response, also induced cross-protection against acid shock conditions (pH 3.5). The acid tolerance response also induced a cross-protection against osmotic shock conditions (20% (w/v) sodium chloride). In some cases there was a relationship between the degree of resistance and adaptation, but usually the behaviour of a particular strain was independent of the conditions from which it was isolated. PMID- 12781944 TI - Simultaneous detection of Escherichia coli O157:H7, Listeria monocytogenes and Salmonella strains by real-time PCR. AB - A protocol enabling simultaneous detection of Escherichia coli O157:H7, Listeria monocytogenes and Salmonella strains was devised and evaluated using artificially contaminated fresh produce. Association of Official Analytical Chemists (AOAC) approved polymerase chain reaction (PCR) detection methods for three human pathogens were modified to enable simultaneous and real-time detection with high throughput capability. The method includes a melting-curve analysis of PCR products, which serves as confirmatory test. The modified protocol successfully detected all three pathogens when fresh produce was washed with artificially contaminated water containing E. coli O157:H7 and S. typhimurium down to the predicted level of 1 to 10 cells/ml and L. monocytogenes at 1000 cells/ml. The ability to monitor several pathogens simultaneously will save time and increase our ability to assure food safety. PMID- 12781945 TI - Isolation and characterization of a Lactobacillus plantarum bacteriophage, phiJL 1, from a cucumber fermentation. AB - A virulent Lactobacillus plantarum bacteriophage, PhiJL-1, was isolated from a commercial cucumber fermentation. The phage was specific for two related strains of L. plantarum, BI7 and its mutant (deficient in malolactate fermenting ability) MU45, which have been evaluated as starter cultures for controlled cucumber fermentation and as biocontrol microorganisms for minimally processed vegetable products. The phage genome of PhiJL-1 was sequenced to reveal a linear, double stranded DNA (36.7 kbp). Sodium dodecyl sulfate-polyacryamide gel electrophoresis (SDS-PAGE) profiles indicated that PhiJL-1 contains six structural proteins (28, 34, 45, 50, 61, and 76 kDa). Electron microscopy revealed that the phage has an isometric head (59 nm in diameter), a long non-contractile tail (182 nm in length and 11 nm in width), and a complex base plate. The phage belongs to the Bradley group B1 or Siphoviridae family. One-step growth kinetics of the phage showed that the latent period was 35 min, the rise period was 40 min, and the average burst size was 22 phage particles/infected cell. Phage particles (90%) adsorbed to the host cells 20 min after infection. Calcium supplementation (up to 30 mM CaCl(2)) in MRS media did not affect the first cycle of phage adsorption, but promoted rapid phage propagation and cell lysis in the infection cycle subsequent to adsorption. The D values of PhiJL-1 at pH 6.5 were estimated to be 2.7 min at 70 degrees C and 0.2 min at 80 degrees C by a thermal inactivation experiment. Knowledge of the properties of L. plantarum bacteriophage PhiJL-1 may be important for the development of controlled vegetable fermentations. PMID- 12781946 TI - Effects of a bacteriocin-like inhibitory substance from Carnobacterium piscicola against human and salmon isolates of Listeria monocytogenes. AB - The aim of this study was to characterize the antagonism of a bacteriocin-like inhibitory substance (BLIS) produced by Carnobacterium piscicola L103 against Listeria monocytogenes strains isolated from salmon and human samples. The inhibitory effect of the BLIS was evaluated in Tryptic soy agar (TSA) during different growth phases of L. monocytogenes at 5 degrees C, using the well diffusion method. Also, the type of inhibition, either bacteriostatic or bactericidal of the BLIS in Tryptic soy broth (TSB), was studied and the development of resistant cells investigated. Results showed an antagonistic effect of the BLIS on all the strains of L. monocytogenes. Four selected strains presented a higher sensitivity to the BLIS in the exponential growth phase and were more resistant in the stationary phase. In TSB, the inhibitory substance showed a partially bactericidal effect on L. monocytogenes. After inactivation of the BLIS with a protease, however, a regrowth of L. monocytogenes was found. The isolate most affected by the action of the BLIS was one of salmon origin. From the 86 isolated colonies that grew in the presence of the BLIS, 93% showed total resistance and 7% partial resistance, which was maintained through five consecutive culture cycles in the absence of the BLIS. PMID- 12781947 TI - Extraction methods for lipopolysaccharides from Escherichia coli ATCC 25922 for quantitative analysis by capillary electrophoresis. AB - Lipopolysaccharides (LPS) are major constituents of the cell wall of Gram negative bacteria. Capillary zone electrophoresis (CZE) was applied to distinguish between lipopolysaccharides extracted from Escherichia coli ATCC 25922 with various techniques. Extraction methods proposed by Westphal and Jann [Methods Carbohydr. Chem. 5 (1965) 83], Galanos et al. [Eur. J. Biochem. 9 (1969) 245], Ni Eidhin and Mouton [FEMS Microbiol. Lett. 110 (1993) 133] and Nichols [Infect. Immun. 62 (1994) 3753] for LPS preparation were evaluated. Electrophoresis buffers with varying pHs were applied to assess the structure stability of the extracted LPS samples. Variations in structural breakdown were apparent demonstrating that different extraction methods removed different LPS molecules. Furthermore, the results obtained proved the CZE useful as an analytical technique for LPS evaluation. The LPS removed with the Nichols extraction procedure presented a unique electrophorogram that could in future be applied in the rapid identification of Gram-negative foodborne pathogens. PMID- 12781948 TI - Proteolytic activities in togwa, a Tanzanian fermented food. AB - Proteolytic activities were investigated in sorghum-based togwa prepared by natural fermentation and using starter cultures previously isolated from the native product, i.e., Lactobacillus brevis, Lactobacillus cellobiosus, Lactobacillus fermentum, Lactobacillus plantarum, Pediococcus pentosaceus, and Issatchenkia orientalis in coculture with either L. brevis or L. plantarum. Both proteinase and aminopeptidase activities were substantially higher in naturally fermented togwa than in those with starters (14-30%, 12-70%, respectively). A variable but substantial part of the proteinase activity followed the particulate fraction of togwa; aminopeptidase activity was mainly in that fraction. The breakdown of relatively high molecular mass protein (64 kDa) in togwa was detected by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS PAGE); the products were mainly in the 14-30 kDa range. Reversed-phase fast protein liquid chromatography (RP-FPLC)-protein/peptide patterns changed during fermentation with some variation between togwa of different cultures. Supplementation of gruel with malt increased the concentration of total protein [from 9.5% to 11.0% (w/w) on dry weight basis)] and of most of the free amino acids. Fermentation had no effect on total protein content; however, the concentration of most of the amino acids was reduced, except for the proline content that increased. Natural fermentation also increased the concentration of glutamic acid and ornithine. Fermentation by P. pentosaceus increased aspartic acid, while L. cellobiosus, L. fermentum, and L. brevis in coculture with I. orientalis increased the concentration of citrulline. PMID- 12781949 TI - Potential osmoprotectants for the lactic acid bacteria Pediococcus pentosaceus and Tetragenococcus halophila. AB - The physiological responses of the lactic acid bacteria Pediococcus pentosaceus and Tetragenococcus halophila (formely known as P. halophila), subjected to osmotic stress in the presence of molecules known to act as osmoprotectants for other bacteria were studied. In a defined medium, glycine betaine, dimethylsulfonioacetate, choline, proline and L-carnitine were able to relieve inhibition of growth at 0.8 M NaCl. The five compounds were shown to efficiently compete with glycine betaine transport, suggesting the existence of common transporter(s) for these molecules. T. halophila, the most tolerant strain, exhibited a larger spectrum of compatible solutes including dimethylsulfonioacetate, dimethylsulfoniopropionate and ectoine. Preliminary data suggest that restoration of growth by ectoine under osmotic constraint seems specific to the genus Tetragenococcus. PMID- 12781950 TI - Yeast autolytic mutants potentially useful for sparkling wine production. AB - A selection method, based on a temperature-sensitive autolytic phenotype, has been used to genetically improve a second fermentation Saccharomyces cerevisiae yeast strain by UV mutagenesis. The mutations carried by the resulting strains affected cell morphology, growth kinetics, sporulation and the release of nitrogenous compounds in an accelerated autolysis experimental model. Their fermentation power was not severely impaired. PMID- 12781951 TI - Presence of macrolide resistance genes in streptococci and enterococci isolated from pigs and pork carcasses. AB - Macrolide and lincosamide (ML)-resistant streptococci and enterococci from tonsillar and colon swabs from 33 pigs and 99 pork carcasses swabs from animals originating from different farms in Belgium were isolated, and their ML resistance phenotypes and genotypes were determined by disk diffusion test and PCR assay, amplifying the ermB gene and the mefA gene. From each of the 33 pigs and 88 of the 99 carcasses' swabs, at least one resistant strain was isolated. The predominant phenotype was the constitutively expressed macrolides, lincosamides and streptogramin B (MLS(B)) phenotype. This phenotype was most often encoded by the ermB gene. A minority of the strains showed the M phenotype encoded by the mefA gene in streptococci, or the L or ML phenotype. PMID- 12781952 TI - Molecular typing techniques as a tool to differentiate non-Saccharomyces wine species. AB - A total of 32 yeast strains belonging to four non-Saccharomyces species associated with winemaking was characterized by different molecular techniques. The PCR amplification of 18S rRNA-coding DNA and nontranscribed spacer, followed by restriction analysis with the endonucleases HaeIII and MspI, and PCR fingerprinting with microsatellite primers (GAC)(5) and (GTG)(5) were used. The methods used provided species-specific profiles and proved to be fast and reliable for monitoring the evolution of the four non-Saccharomyces yeast populations throughout wine fermentation. PMID- 12781953 TI - Characterisation of Aeromonas spp. isolated from frozen fish intended for human consumption in Mexico. AB - A total of 82 strains of presumptive Aeromonas spp. were identified biochemically and genetically (16S rDNA-RFLP). The strains were isolated from 250 samples of frozen fish (Tilapia, Oreochromis niloticus niloticus) purchased in local markets in Mexico City. In the present study, we detected the presence of several genes encoding for putative virulence factors and phenotypic activities that may play an important role in bacterial infection. In addition, we studied the antimicrobial patterns of those strains. Molecular identification demonstrated that the prevalent species in frozen fish were Aeromonas salmonicida (67.5%) and Aeromonas bestiarum (20.9%), accounting for 88.3% of the isolates, while the other strains belonged to the species Aeromonas veronii (5.2%), Aeromonas encheleia (3.9%) and Aeromonas hydrophila (2.6%). Detection by polymerase chain reaction (PCR) of genes encoding putative virulence factors common in Aeromonas, such as aerolysin/hemolysin, lipases including the glycerophospholipid cholesterol acyltransferase (GCAT), serine protease and DNases, revealed that they were all common in these strains. Our results showed that first generation quinolones and second and third generation cephalosporins were the drugs with the best antimicrobial effect against Aeromonas spp. In Mexico, there have been few studies on Aeromonas and its putative virulence factors. The present work therefore highlights an important incidence of Aeromonas spp., with virulence potential and antimicrobial resistance, isolated from frozen fish intended for human consumption in Mexico City. PMID- 12781954 TI - Inactivation of Lactobacillus delbrueckii bacteriophages by heat and biocides. AB - The effect of several biocides and thermal treatments on the viability of four Lactobacillus delbrueckii phages was investigated. Time to achieve 99% inactivation of phages at 63 and 72 degrees C in three suspension media (Tris Magnesium Gelatin (TMG) buffer, Man Rogosa Sharpe (MRS) broth and reconstituted nonfat dry skim milk (RSM)) was calculated. Thermal resistance depended on the phage considered, but a marked heat-resistance was exhibited by one phage (Ib(3)) since its high titre suspensions were completely inactivated only after 45 min at 72 degrees C or 15 min at 90 degrees C. A clear protective effect of the milk was revealed when the three suspension media were compared. As regards to the effects of biocides on phages, only peracetic acid was found to be effective for inactivating high titre suspensions. Ethanol, even at a concentration of 100%, was not suitable to assure no surviving phage particles and isopropanol turned out to be less effective than ethanol. Sodium hypochlorite at 200-400 ppm inactivated the phages completely, except phage Ib(3), which was only destroyed after treatments with 1200 ppm. The diversity observed in the heat and biocide resistance of L. delbrueckii phages is useful to establish a basis for adopting the most effective thermal and chemical treatments for inactivating them in dairy plants and laboratory environments. PMID- 12781955 TI - Isolation and molecular characterization of Escherichia coli O157 isolated from cattle, pigs and chickens at slaughter. AB - From 1999 until 2001, 3625 food samples were examined for the presence of Escherichia coli O157. Samples were from bovine origin (ground beef, n=549; carcasses, n=2452), calves (carcasses, n=147), chicken (breast, n=203; carcasses, n=71) and pigs (carcasses, n=85; trimmings, n=118). Vidas ECO detected 451 (12%) samples positive, but from only 27 (0.74%) samples was E. coli O157 isolated. One strain was isolated from bovine ground beef (0.18%), one from a pig carcass (1.17%) and all others were isolated from bovine carcasses (1.02%). All strains possessed the attaching-and-effacing gene, the enterohemorrhagic plasmid and verotoxin (VT) genes, except the strain isolated from the pig carcass that was therefore eliminated. Six of the strains were urease-positive. Strains were typed by two DNA fingerprinting methods: random amplification of polymorphic DNA (RAPD) and pulsed field gel electrophoresis (PFGE). PFGE revealed a similarity of 71.05%, while RAPD was 77.36% similar. None of the typing methods were able to classify all urease-positive strains to one pattern. Strains in the same PFGE cluster did not belong to one RAPD cluster. This paper highlights that Belgian fresh meat at retail level can be contaminated with E. coli O157 and that two different typing methods divide strains into different types. PMID- 12781956 TI - Molecular differentiation of clostridia associated with "blown pack" spoilage of vacuum-packed meats using internal transcribed spacer polymorphism analysis. AB - The 16S-23S rDNA internal transcribed spacer (ITS) polymorphism analysis was assessed for its suitability in rapid discrimination between species of psychrophilic and psychrotolerant clostridia associated with "blown pack" spoilage of vacuum-packed meats. DNA isolated from 10 reference and 20 meat strains of psychrophilic and psychrotolerant clostridia were used as templates in PCR amplification with primers complementary to conserved regions of the 3' end of the 16S rRNA and 5' end of the 23S rRNA genes directly flanking the spacer. The majority of strains showed multi-band ITS patterns when products of spacer amplification were visualised on an agarose gel. With the majority of meat strains, PCR amplification generated single banding pattern for a single clostridial species. However, meat strains of Cl. algidicarnis produced four different ITS banding patterns. With reference strains of psychrophilic and psychrotolerant clostridia, variation in spacer length was also observed between nonproteolytic Cl. botulinum type B (17B), E (Beluga) and F (202F). On the other hand, the number and size of the ITS amplification products could not be used for a differentiation of Cl. laramiense ATCC 51254(T) from Cl. estertheticum DSM 8809(T), Cl. putrefaciens DSM 1291(T) from Cl. algidicarnis NCFB 2931(T), or Cl. frigidicarnis strains from nonproteolytic Cl. botulinum type B (17B). The presence of interstrain, and lack of interspecies, ITS polymorphism observed in the present study with some clostridial species may preclude the use of 16S-23S rDNA spacer amplification for species-level discrimination and identification, respectively, of psychrophilic and psychrotolerant clostridia associated with meat spoilage. However, where interstrain, intraspecies heterogeneity of ITS amplification products exists, ITS analysis could be useful for tracing back psychrophilic and psychrotolerant clostridia responsible for meat spoilage to their meat plant sources. PMID- 12781957 TI - Rapid enumeration of Listeria monocytogenes in milk using competitive PCR. AB - Competitive polymerase chain reaction (cPCR) was used to develop a direct enumeration method of Listeria monocytogenes in milk. Sterile milk was artificially inoculated with L. monocytogenes and DNA was extracted using guanidine thiocyanate/phenol/chloroform, followed by PCR. Several primers for L. monocytogenes hlyA gene were tested for specific detection and DG69/DG74 primer set was selected. The primer set produced a 636-bp band from L. monocytogenes, but no band appeared from the other six Listeria spp. tested. A detection limit was as few as 10(3) colony-forming unit (cfu) per 0.5 ml of milk with this primer set. When the samples were cultured at 25 degrees C for 15 h in a TSBY medium, even a single bacterium could be detected with this primer set by PCR. For the cPCR, hlyA gene segment was cloned in pGem-4Z vector and was modified to produce competitor DNA. The competitor DNA has the same primer binding sites and sequences as the target DNA except EcoRI site. Known amount of competitor DNA was coamplified with L. monocytogenes total DNA isolated from artificially inoculated milk. The target DNA and competitor DNA were distinguished by EcoRI digestion after cPCR. The cell number determined by cPCR was approximately equal to the colony-forming unit from conventional plate counting method. For the whole procedure, it took only 5 h. PMID- 12781958 TI - Antimicrobial-resistant Salmonella serovars isolated from imported foods. AB - A total of 187 Salmonella isolates representing 82 serotypes recovered from 4072 imported foods in the year 2000 by the U.S. Food and Drug Administration field laboratories were tested for their susceptibility to 17 antimicrobials of human and veterinary importance. Fifteen (8%) isolates were resistant to at least one antimicrobial, and five (2.7%) were resistant to three or more antimicrobials. Most of the isolates (n=9) exhibited resistance to tetracycline. Four isolates from catfish or tilapia from Taiwan or Thailand also demonstrated resistance to nalidixic acid. These nalidixic acid-resistant Salmonella isolates possessed a point mutation at the Ser83 or Asp87 position in DNA gryase, resulting in amino acid substitutions to phenylalanine, tyrosine, or asparagine. One Salmonella Derby isolated from frozen anchovies imported from Cambodia was resistant to six antimicrobials including ampicillin, amoxicillin/clavulanic acid, chloramphenicol, sulfamethoxazole, tetracycline, and trimethoprim/sulfamethoxazole. Of seven isolates displaying resistance to sulfonamides, only one S. Derby and one Salmonella Agona contained class 1 integrons that were further shown to possess the aadA and pse-1 genes conferring resistance to streptomycin and ampicillin, respectively. This study indicates that antimicrobial-resistant Salmonella are present in imported foods, primarily of seafood origin, and stresses the need for continued surveillance of foodborne zoonotic bacterial pathogens from imported foods entering the United States. PMID- 12781959 TI - Biological control of Monilinia laxa and Rhizopus stolonifer in postharvest of stone fruit by Pantoea agglomerans EPS125 and putative mechanisms of antagonism. AB - Treatment of stone fruits (apricot, peach and nectarine) with Pantoea agglomerans strain EPS125 decreased the incidence and diameter of lesions of brown rot caused by Monilinia laxa and soft rot caused by Rhizopus stolonifer. Root control was achieved on fruits either wounded and subsequently inoculated with the pathogens or non-wounded and naturally infected from orchards. The efficacy of biocontrol was dependent on the concentration of the biocontrol agent and pathogen. At medium to low pathogen dose, optimal EPS125 concentrations were above 10(7) CFU ml(-1). The median effective dose (ED(50)) of EPS125 was 4.5x10(4) in M. laxa and 2.2x10(5) CFU ml(-1) in R. stolonifer. However, EPS125 was more effective in M. laxa than in R. stolonifer as indicated by the ratio between ED(50) of the biocontrol agent and pathogen (K(z)/K(x)) which was 166 and 1263, respectively. Interactions between the strain EPS125 and the fruit surface, and M. laxa and R. stolonifer, were studied to determine the mechanisms of protection from postharvest rots. The strain EPS125 colonizes, grows and survives on stone fruit wounds. Significant inhibition of conidial germination and hyphal growth of R. stolonifer and M. laxa was achieved when the fungal and EPS125 cells were cocultivated on peel leachate or nectarine juice. However, no effect was observed when the antagonist and the pathogen cells were physically separated by a membrane filter which permits nutrient and metabolite interchange. Therefore, a direct interaction between the strain and the pathogen cells is necessary for antagonism, without a significant contribution of the production of antibiotic substances or nutrient competition. Preemptive exclusion by wound colonization and direct interaction with the pathogen is proposed as the mechanism of biocontrol. PMID- 12781960 TI - Campylobacter jejuni contamination on broiler carcasses of C. jejuni-negative flocks during processing in a Japanese slaughterhouse. AB - At a slaughterhouse, we determined the Campylobacter jejuni status of each broiler flock coming from different farms by culturing cecal contents of representative birds, and examined the prevalence of C. jejuni on carcasses during processing. Then, C. jejuni isolates from the carcasses were typed with RAPD PCR and compared with those from the cecal contents of a C. jejuni-positive flock. C. jejuni was not isolated from the carcasses of C. jejuni-negative flocks processed before the C. jejuni-positive flock, and C. jejuni was isolated from the carcasses of C. jejuni-negative flocks processed after the C. jejuni-positive flock. RAPD PCR type of the isolates from the carcasses of C. jejuni-negative flocks was the same as those from the cecal contents of the previously processed C. jejuni-positive flock. Our results suggest that the carcasses of the C. jejuni negative flocks were contaminated with C. jejuni strains originating from the intestines of the previously processed C. jejuni-positive flock. PMID- 12781961 TI - Rapid urease screening of Yersinia on CIN agar plates. AB - Yersinia enterocolitica is an important foodborne pathogen, but isolation of virulent Yersinia from food sources is still time consuming and requires skills. In this article, we describe a rapid urease screening on cefsulodin-irgasan novobiocin (CIN) agar plates with an agar overlay assay. This test is simple to perform, all colonies on a plate can be checked simultaneously, it only takes minutes for detection of urease-positive colonies and the colonies survive for transfer, further characterisation, and storage. Additionally, this method is useful to isolate virulent (urease-positive and pYV harbouring) Y. enterocolitica from foodstuffs. PMID- 12781962 TI - Screening of biogenic amine production by lactic acid bacteria isolated from grape must and wine. AB - The potential to produce the biogenic amines tyramine, histamine and putrescine, was investigated for lactic acid bacteria (LAB) of various origin, including commercial malolactic starter cultures, type strains and 78 strains isolated from Spanish grape must and wine. The presence of biogenic amines in a decarboxylase synthetic broth was determined by reverse-phase high performance liquid chromatography (RP-HPLC). Tyramine was the main amine formed by the LAB strains investigated. Leuconostoc strains were the most intensive tyramine formers. No potential to form biogenic amines was observed in Oenococcus oeni strains. Two strains of Latobacillus buchneri were associated with putrescine formation. None of the lactic acid bacteria produced histamine. According to these in vitro results, the commercial starter bacteria analyzed did not produce histamine, tyramine and putrescine. PMID- 12781966 TI - Mechanism of cationic surfactant adsorption at the solid-aqueous interface. AB - Until recently, the rapid time scales associated with the formation of an adsorbed surfactant layer at the solid-aqueous interface has prevented accurate investigation of adsorption kinetics. This has led to the mechanism of surfactant adsorption being inferred from thermodynamic data. These explanations have been further hampered by a poor knowledge of the equilibrium adsorbed surfactant morphology, with the structure often misinterpreted as simple monolayers or bilayers, rather than the discrete surface aggregates that are present in many surfactant-substrate systems. This review aims to link accepted equilibrium data with more recent kinetic and structural information in order to describe the adsorption process for ionic surfactants. Traditional equilibrium data, such as adsorption isotherms obtained from depletion approaches, and the most popular methods by which these data are interpreted are examined. This is followed by a description of the evidence for discrete aggregation on the substrate, and the morphology of these aggregates. Information gained using techniques such as atomic force microscopy, fluorescence quenching and neutron reflectivity is then reviewed. With this knowledge, the kinetic data obtained from relatively new techniques with high temporal resolution, such as ellipsometry and optical reflectometry, are examined. On this basis the likely mechanisms of adsorption are proposed. PMID- 12781965 TI - Surface forces in wetting films. AB - A short review of various components of surface forces acting in a non symmetrical system such as wetting films is presented here. Experimental results are compared with modified DLVO theory, which includes, besides dispersion and electrostatic, structural (solvation) forces caused by a change in liquid structure in conditions of confined geometry. The peculiarities of disjoining pressure isotherms and conditions of the film stability of non-polar and polar simple liquids, as well as of aqueous solutions of electrolytes and surfactants, are systematically considered from a historical perspective. PMID- 12781967 TI - The water-soluble matrix fraction from the nacre of Pinctada maxima produces earlier mineralization of MC3T3-E1 mouse pre-osteoblasts. AB - Nacre or mother of pearl is a calcified structure that forms the lustrous inner layer of some shells. We studied the biological activity of the water-soluble matrix (WSM) extracted from powdered nacre from the shell of the pearl oyster, Pinctada maxima, on the MC3T3-E1 pre-osteoblast cell line from mouse calvaria. This cell line has the ability to differentiate into osteoblasts and to mineralize in the presence of beta-glycerophosphate and ascorbic acid. Cell proliferation and alkaline phosphatase activity were measured as markers of osteoblast differentiation, and mineralization was analyzed. These studies revealed that WSM stimulates osteoblast differentiation and mineralization by day 6 instead of the 21-day period required for cells grown in normal mineralizing media. We compared the activity of WSM with that of dexamethasone on this cell line. WSM can inhibit alkaline phosphatase (ALP) activity and the activity of dexamethasone on MC3T3-E1 cells. This study shows that nacre WSM could speed up the differentiation and mineralization of this cell line more effectively than dexamethasone. PMID- 12781968 TI - Na+/Ca2+ exchanger-deficient mice have disorganized myofibrils and swollen mitochondria in cardiomyocytes. AB - The Na(+)/Ca(2+) exchanger (NCX1) plays a key role in maintaining Ca(2+) homeostasis in cardiomyocytes. Disruption of Ncx1 gene in mice results in embryonic lethality between embryonic day 9 and 10, with the mice lacking spontaneous heartbeats. We examined the mechanism of lack of heartbeats in Ncx1 deficient mice. Ultrastructual analysis demonstrated that Ncx1-deficient mice showed severe disorganization of myofibrils, a lack of Z-lines and swelling of mitochondria in cardiomyocytes. However, the expressions of cardiac-specific genes including transcription factor genes and contractile protein genes were not changed in Ncx1-deficient mice. Abnormal Ca(2+) handling itself or the lack of heartbeats due to the inactivation of Ncx1 gene may cause the disorganization of myofibrillogenesis. Although NCX1 protein levels were decreased in heterozygous mice, there were no changes in NCX2 and NCX3 protein levels between wild type and heterozygous mice. PMID- 12781969 TI - Characterization of metalloproteinase-like activities in barnacle (Balanus amphitrite) nauplii. AB - The presence of extracellular matrix (ECM) degrading enzymes was investigated in naupliar stages of the barnacle Balanus amphitrite Darwin. The results of substrate gel-zymography and quantitative assays demonstrated that naupliar extracts contain several protease activities that are specific towards gelatin substrates; some caseinolytic activity was also detected. Substrate specificity was observed in all naupliar stages (II-VI). The gelatinolytic activities showed dependence on both Ca(2+) and Zn(2+) and inhibition by EDTA, EGTA, and 1,10 phenanthroline. Also Mg(2+) partially activated the enzymes, whereas Cd(2+), Cu(2+), Hg(2+) and Pb(2+) were inhibitory. The thermal denaturation profile was significantly different in the presence and absence of Ca(2+) and Zn(2+). Overall, the results indicate that the Ca(2+)/Zn(2+)-dependent gelatinase activities in barnacle nauplii belong to the subfamily of matrix metalloproteases. Barnacle larvae MMPs showed biochemical characteristics different from those of vertebrate MMPs but common to other gelatinases from marine invertebrates: they were unaffected by several protease inhibitors and insensitive to specific activators/inhibitors of vertebrate MMPs. The presence of MMP-like activities in different naupliar stages suggests a constitutive role for these enzymes in ECM remodeling during barnacle larvae growth and development. PMID- 12781970 TI - The phylogenetic distribution of sphingomyelinase D activity in venoms of Haplogyne spiders. AB - The venoms of Loxosceles spiders cause severe dermonecrotic lesions in human tissues. The venom component sphingomyelinase D (SMD) is a contributor to lesion formation and is unknown elsewhere in the animal kingdom. This study reports comparative analyses of SMD activity and venom composition of select Loxosceles species and representatives of closely related Haplogyne genera. The goal was to identify the phylogenetic group of spiders with SMD and infer the timing of evolutionary origin of this toxin. We also preliminarily characterized variation in molecular masses of venom components in the size range of SMD. SMD activity was detected in all (10) Loxosceles species sampled and two species representing their sister taxon, Sicarius, but not in any other venoms or tissues surveyed. Mass spectrometry analyses indicated that all Loxosceles and Sicarius species surveyed had multiple (at least four to six) molecules in the size range corresponding to known SMD proteins (31-35 kDa), whereas other Haplogynes analyzed had no molecules in this mass range in their venom. This suggests SMD originated in the ancestors of the Loxosceles/Sicarius lineage. These groups of proteins varied in molecular mass across species with North American Loxosceles having 31-32 kDa, African Loxosceles having 32-33.5 kDa and Sicarius having 32-33 kDa molecules. PMID- 12781971 TI - Carotenoprotein complexes in entomostracan crustaceans (Streptocephalus dichotomus and Moina micrura). AB - The carotenoprotein complexes of a freshwater fairy shrimp (Streptocephalus dichotomus) and a daphnid (Moina micrura) were characterised and compared. Based on thin layer chromatography and mass spectral analysis, a variety of cartenoprotein complexes such as astaxanthin, canthaxanthin, antheraxanthin, lutein, beta-cryptoxanthin and violaxanthin were found. Both crustaceans had astaxanthin and canthaxanthin as predominant prosthetic groups. Amino acid analysis of the complexes further revealed high levels of asparagine, glutamine and glycine in both species. Our study highlights the presence of naturally available carotenoid species in both crustaceans and their possible inter conversion in anostracans. PMID- 12781972 TI - A novel ferritin subunit involved in shell formation from the pearl oyster (Pinctada fucata). AB - Iron is one of the most important minor elements in the shell of bivalves. This study was designed to investigate the involvement of ferritin, the principal protein for iron storage, in shell formation. A novel ferritin cDNA from the pearl oyster (Pinctada fucata) was isolated and characterized. The ferritin cDNA encodes a 206 amino acid polypeptide, which shares high similarity with snail soma ferritin and the H-chains of mammalian ferritins. Oyster ferritin mRNA shows the highest level of expression in the mantle, the organ for shell formation. In situ hybridization analysis revealed that oyster ferritin mRNA is expressed at the highest level at the mantle fold, a region essential for metal accumulation and contributes to metal incorporation into the shell. Taken together, these results suggest that ferritin is involved in shell formation by iron storage. The identification and characterization of oyster ferritin also helps to further understand the structural and functional properties of molluscan ferritins. PMID- 12781973 TI - Acetylcholinesterase and butyrylcholinesterase activities in brain and plasma of freshwater teleosts: cross-species and cross-family differences. AB - Brain and plasma acetylcholinesterase (AChE; EC 3.1.1.7) and plasma butyrylcholinesterase (BChE; EC 3.1.1.8) specific activities were assayed in 16 freshwater teleosts belonging to four families: Cyprinidae, Percidae, Esocidae and Lotidae. Brain AChE activity varied among fish species approximately 15-fold, ranging from 138 to 2011 micromol/g per h. All cyprinids had higher brain AChE activity than other fish families. Plasma AChE activity was on average 100-fold lower than that in brain, varying from 1.2 to 18.6 micromol/ml per h. Plasma BChE activity was found only in cyprinids with the exception of the common and crucian carp, and sabrefish. It varied from 26 to 1083 micromol/ml per h. In bream (Abramis brama) only 30% of specimens studied had BChE activity. The correlation coefficient values between activities of brain and plasma AChE, brain AChE and plasma BChE, plasma AChE and BChE were 0.67, 0.68 and 0.84, respectively. Cross species and also cross-family differences in AChE and BChE activities among fish were demonstrated. Possible reasons for the differences are discussed. PMID- 12781974 TI - Expression of alpha-amylase isozymes in rat tissues. AB - Gene expressions of alpha-amylase isozymes in rat tissues were analyzed by a reverse transcription-polymerase chain reaction (RT-PCR), followed by EcoRI digestion. This procedure is based on evidence that an RT-PCR product from mouse pancreas RNA is sensitive to EcoRI, but not the product from the salivary gland or liver RNAs. The method was applied to the analysis of alpha-amylase expression in rat liver after partial hepatectomy, in which a potent expression of pancreas type isozyme was observed. However, no expression of the pancreatic isozyme in the regenerating liver was found. We also analyzed the expression of alpha amylase gene in several additional rat tissues. In intestine, stomach, testis and skeletal muscle, the corresponding PCR products were amplified, but few were detected in heart or spleen. Intestine and stomach expressed a pancreatic isozyme of alpha-amylase. Analyses of the alpha-amylase activity and protein indicated the presence of the enzyme in those tissues. Immunohistochemical analysis also indicated that the amylase proteins were specifically present in epithelial cells of rat intestinal mucosa. This is a convenient method for identification of alpha amylase isozyme mRNA in rodent tissues. PMID- 12781975 TI - Cleavage site of a major yolk protein (MYP) determined by cDNA isolation and amino acid sequencing in sea urchin, Hemicentrotus pulcherrimus. AB - The overall sequence of cDNA encoding vitellogenin (Vg), a precursor to major yolk protein (MYP), of Hemicentrotus pulcherrimus was determined. Its nucleotide sequence has an open reading frame of 4041 bp encoding 1346 amino acids. The amino acid sequence showed little similarity to other Vgs in vertebrates, insects or nematodes, but resembled members of the vertebrate and invertebrate transferrin family. The N-terminal amino acid sequence of the protein fragments dominant in the later embryonic stage was analyzed in order to determine the cleavage site of MYP. Determination of the cleavage site in MYP and analysis of MYP proteolysis in vitro suggested that MYP has a specific molecular shape to permit its proteolytic fragmentation at a definite site. The functional region of transferrin in MYP is conserved after proteolytic processing. Considering these results and those from other work, the protein called sea urchin Vg is not a true Vg. Therefore, a new name, echinoferrin, is proposed for this protein. PMID- 12781976 TI - Tissue essential fatty acid composition and competitive response to dietary manipulations in white bass (Morone chrysops), striped bass (M. saxatilis) and hybrid striped bass (M. chrysopsxM. saxatilis). AB - The effects of wide changes in dietary levels of docosahexaenoic (DHA) or arachidonic (ArA) acids on growth, survival and fatty acid composition in body tissues of Morone larvae were examined. White bass (WB, Morone chrysops), striped bass (SB, Morone saxatilis) and sunshine hybrid bass (HSB, M. chrysopsxM. saxatilis) larvae (day 24-46) were fed Artemia nauplii enriched with algal sources of varying proportions of DHA and ArA (from 0 to over 20% of total fatty acids). WB larvae fed DHA-deficient Artemia diet retarded over 50% of their potential growth, however, increasing dietary DHA/ArA ratios were associated with a significant growth improvement. The highest proportion of polyunsaturated fatty acids was found in WB neural tissue (approx. 50% of total fatty acids), while HSB neural tissue contained the highest proportion of saturated fatty acids (approx. 35% of total fatty acids). Within the neural tissues of all Morone larvae, both DHA and ArA were generally the most dominant as well as the most responding fatty acids to dietary manipulations (except in WB fed DHA or ArA deficient diets). HSB neural tissue was particularly efficient in retaining a significant amount of DHA in the face of dietary deficiency. However, WB neural tissue was the most responsive to dietary increase in DHA, accumulating a significantly higher amount of DHA (P<0.05) than SB or HSB. Results demonstrate significant differences in fatty acid composition and growth responsiveness to dietary manipulations between Morone larvae species and within specific tissues. WB weight gain and neural tissue composition was affected most by dietary changes in both DHA and ArA whereas SB and HSB tissue compositions were generally less affected by dietary manipulations. PMID- 12781977 TI - Effect of dietary lipid level on fatty acid beta-oxidation and lipid composition in various tissues of haddock, Melanogrammus aeglefinus L. AB - Haddock (Melanogrammus aeglefinus) is a gadoid fish species that deposits dietary lipid mainly in the liver. The fatty acid (FA) beta-oxidation activity of various tissues was evaluated in juvenile haddock fed graded levels of lipid. The catabolism of a radiolabelled FA, [1-(14)C]palmitoyl-CoA, through peroxisomal and mitochondrial beta-oxidation was determined in the liver, red and white muscle of juvenile haddock fed 12, 18 and 24% lipid in the diet. There was no significant increase in the mitochondrial or peroxisomal beta-oxidation activity in the tissues tested as the dietary lipid level increased from 12 to 24%. Peroxisomes accounted for 100% of the beta-oxidation observed in the liver, whereas mitochondrial beta-oxidation dominated in the red (91%) and white muscle (97%) of juvenile haddock. Of the tissues tested, red muscle possessed the highest specific activity for beta-oxidation expressed on a per mg protein or per g wet weight basis. However, white muscle, which forms over 50% of the body mass in gadoid fish was the most important tissue in juvenile haddock for overall FA catabolism. The total lipid and FA composition of these tissues were also determined. This study confirmed that the liver was the major lipid storage organ in haddock. The hepatosomatic index (HSI; 10.0-15.2%) and lipid (73.8-79.3% wet wt.) in the liver increased significantly as dietary lipid was increased from 12 to 24% lipid. There was no significant increase in the lipid composition of the white muscle (0.8% wet wt.), red muscle (1.9% wet wt.) or heart (2.5% wet wt.). PMID- 12781978 TI - cDNA cloning of growth hormone from giant panda (Ailuropoda melanoleuca) and its expression in Escherichia coli. AB - A cDNA encoding Ailuropoda melanoleuca growth hormone (AmGH) was isolated from pituitary total RNA using RT-PCR and expressed in Escherichia coli. This is the first report of a GH nucleotide and amino acid (aa) sequence from giant panda. The open reading frame of AmGH (651 bp) encodes a precursor of 216 aa comprising a 26 aa signal peptide and a 190 aa mature protein with four cysteine residues similar to the typical primary structure of mammalian GH precursor. AmGH shares a high degree of identity (54-98.9%) with that of mammals, birds and amphibians, but a very low identity with bony fish GH (only 20-30%). The mature AmGH exhibits striking similarity to that of putative ancestral GH with a difference of only two residues, indicating a very slow basal rate of molecular evolution. The DNA fragment encoding mature AmGH was then subcloned into the pGEX-4T-1 expression vector and highly expressed in E. coli host BL21 with IPTG induction. The expressed proteins fused to GST were found to be sequestered into inclusion bodies and therefore the NaOH method was employed to solubilize the inclusion bodies; the proteins were further purified by Glutathione Sepharose 4B affinity chromatography. The production and purification of GST-AmGH reported here provide a basis for further studies on the biological activity of AmGH. PMID- 12781979 TI - cDNA cloning of a defender against apoptotic cell death 1 (DAD1) homologue, responsive to external temperature stimulus from the spider, Araneus ventricosus. AB - A cDNA encoding a defender against apoptotic cell death (DAD1) homologue was cloned from a cDNA library of the spider, A. ventricosus. Sequence analysis of the cDNA encoding the DAD1 homologue of A. ventricosus revealed that the 339-bp cDNA has an open reading frame of 113 amino acid residues. The deduced amino acid sequence of the A. ventricosus DAD1 homologue showed 75.4% of identity to Drosophila melanogaster, 74.6% of identity to Xenopus laevis, and 73.1% of identity to Homo sapiens, Sus scrofa, Mesocricetus auratus, Rattus norvegicus and Mus musculus. All animal DAD1 including A. ventricosus DAD1 homologue formed a subgroup, excluding all plant DAD1 proteins in the phylogenetic analysis. Northern blot analysis shows that the transcripts of A. ventricosus DAD1 homologue gene are present in all tissues examined, suggesting that A. ventricosus DAD1 is expressed in most, if not all, body tissues. Interestingly, the transcript levels of A. ventricosus DAD1 homologue gene were particularly high when exposed at low (4 degrees C) and high (37 degrees C) temperatures, suggesting that the gene is responsive to temperature stimuli. PMID- 12781980 TI - Expression of a midgut-specific cadherin BT-R1 during the development of Manduca sexta larva. AB - The btr-1 gene of Manduca sexta (GenBank AF319973) encodes a cadherin, BT-R(1) (210-kDa), which contains 12 ectodomain modules in association with a number of motifs potentially involved in interactions with cadherin and integrin. The molecule is a target receptor for Bacillus thuringiensis Cry1A toxins that bind to BT-R(1) with high affinity and specificity. BT-R(1) is localized exclusively in the midgut epithelium. The amount of BT-R(1) protein increases dramatically during larval development, paralleling accumulation of its mRNA. The 5'-UTR of the btr-1 gene contains sequence motifs that most likely recruit specific transcription factors, particularly, those that determine posterior patterning and that control intestinal cell proliferation, differentiation and identity during development. The increase in abundance of BT-R(1) may be required to support not only the differentiation of the epithelial cells but also the establishment of physiological function and structural integrity of the midgut during larval development in M. sexta. We believe that BT-R(1) is essential to larval midgut epithelial organization during rapid cell proliferation and tissue growth in M. sexta because disruption of such organization and functionality occasioned by the binding of the Cry1A toxins of B. thuringiensis to BT-R(1) causes death to the insect. PMID- 12781981 TI - Cloning and tissue distribution of a carnitine palmitoyltransferase I gene in rainbow trout (Oncorhynchus mykiss). AB - The carnitine palmitoyltransferase I (EC.2.3.1.21; CPT I) mediates the transport of fatty acids across the outer mitochondrial membrane. In mammals, there are two different proteins CPT I in the skeletal muscle (M) and liver (L) encoded by two genes. The carnitine palmitoyltransferase system of lower vertebrates received little attention. With the aim of improving knowledge on the CPT family in fish, we examined CPT I cDNA and CPT activity in different tissues of rainbow trout (Oncorhynchus mykiss). Using RT-PCR, we successfully cloned a partial CPT I cDNA sequence (1650 bp). The predicted protein sequence revealed identities of 63% and 61% with human L-CPT I and M-CPT I, respectively. This mRNA is expressed in liver, white and red skeletal muscles, heart, intestine, kidney and adipose tissue of trout. This is in good agreement with the measurement of the CPT activity in the same tissues. The [IC(50)] that reflects the sensitivity to malonyl-CoA inhibition was 0.116+/-0.004 microM for the liver and 0.426+/-0.041 microM for the white muscle. These results demonstrate for the first time the existence of at least one gene encoding for CPT I present in both the liver and the muscle of rainbow trout. PMID- 12781982 TI - Morphology and chemistry of Dufour glands in four ectoparasitoids: Cephalonomia tarsalis, C. waterstoni (Hymenoptera: Bethylidae), Anisopteromalus calandrae, and Pteromalus cerealellae (Hymenoptera: Pteromalidae). AB - The venom apparatus of four hymenopterous parasitoids, including two bethylids, C. tarsalis (Ashmead) and C. waterstoni (Gahan), and two pteromalids, A. calandrae (Howard) and P. cerealellae (Ashmead), were removed and the associated Dufour glands characterized with respect to their external morphology and chemistry. Dufour glands in all four species have a characteristic translucent appearance that apparently results from their lipid content. The stalked Dufour glands of C. tarsalis and C. waterstoni are pear-shaped and have overall lengths of approximately 0.2 and 0.15 mm, respectively. The thin venom glands are bifurcate and insert through a fine duct into the transparent ovoid- to pear shaped venom reservoir in these bethylids. In A. calandrae and P. cerealellae the Dufour glands are elongated, tubular structures of ca. 0.35 and 0.8 mm in length, respectively, that constrict to a short stalk that empties into the common oviduct. The venom glands in these pteromalids are simple elongated structures that insert into the sac-like venom reservoir through a fine duct. The chemistry of the volatile contents of the Dufour gland in these four species differs considerably. C. tarsalis Dufour glands contain the same hydrocarbon components as found on the cuticle of this species (Ann. Entomol. Soc. Am. 91:101-112 (1998)), and no other chemicals. The Dufour glands of C. waterstoni also contain only hydrocarbons, most of which are the same as the cuticular hydrocarbons (Ann. Entomol. Soc. Am. 85:317-325 (1992)), but in addition the Dufour gland contains ca. 3% of a mixture of 2,17- and 2,19-dimethyl C(23). A. calandrae Dufour gland chemistry is somewhat more complex than that of either of the two bethylids, but like the bethylids, only hydrocarbons are present. The carbon number range is from C(30) to C(39) and consists of a mixture of n-alkanes (C(30)-C(38)); 3-, 5-, 7-, 9-, 11-, 12-, 13-, 14-, 15- and 17-methyl alkanes; 3,7- and 3,11-dimethyl alkanes; 5,9- and 5,17-dimethyl alkanes; 7,11-, 9,13-, 13,17-, 14,18- and 15,19 dimethyl alkanes; 3,7,11- and 3, 9,15-trimethyl alkanes; and 3,7,11,15 tetramethyl alkanes. The cuticular hydrocarbons of this species have not been previously reported, but they are the same as the Dufour gland hydrocarbons. The Dufour glands of P. cerealellae contain both hydrocarbons and two long-chain aldehydes. Most of the hydrocarbons are identical to those found on the cuticle of this species (Ann. Entomol. Soc. Am. 94:152-158 (2001)), but in addition, 5,9 dimethyl C(27), 5,13-, 5,17- and 5,19-dimethyl C(35), 12- and 14-methyl C(36), 12,16- and 13,17-dimethyl C(36), 13-methyl C(37) and 13,17-dimethyl C(37) are present. The two aldehydes detected in glands from P. cerealellae are n tetracosanal (C(23)CHO) and n-hexacosanal (C(25)CHO). PMID- 12781983 TI - Analysis of highly phosphorylated inositols in avian and crocodilian erythrocytes. AB - Both morphological and paleontological characteristics support the hypothesis of a monophyletic origin of crocodilian and avian groups. However, while the erythrocytes of all birds studied to date are reported to contain high levels of inositol pentakisphosphate (InsP(5)), which acts as an allosteric effector of hemoglobin, this molecule has not been reported in crocodilian erythrocytes. In this study we compare the highly phosphorylated inositols in crocodilian and avian erythrocytes using a particularly sensitive analytical procedure. Our aim was to obtain new data which might provide further evidence for the monophyletic origin, or otherwise, of crocodiles and birds. We studied three avian and three crocodilian species. The erythrocytes of the three bird species contained low levels of inositol-3,4,5,6-tetrakisphosphate and inositol-1,3,4,6 tetrakisphosphate, thought to be precursors of Ins(1,3,4,5,6)P(5). The crocodilian erythrocytes studied contained Ins(1,3,4,5,6)P(5) and InsP(6) in higher concentrations than those found in mammal erythrocytes and in other more active cells such as macrophages. Our data provide further evidence of the similarity between crocodilian and avian groups and agree with the hypothesis that both groups evolved from a common ancestor. The process by which the function of inositol phosphates changed from that of intracellular signaling to hemoglobin allosteric effector is discussed. PMID- 12781984 TI - Thyroid hormone may induce changes in the concentration of the mitochondrial calcium uniporter. AB - We explored the possibility that the hormone 3,3',5-tri-iodothyronine can regulate the biosynthesis of the mitochondrial calcium uniporter. To meet this objective experiments on Ca(2+) transport, and binding of the specific inhibitor Ru(360) were carried out in mitochondria isolated from euthyroid, hyperthyroid and hypothyroid rats. It was found that V(max) for Ca(2+) transport increased from 11.67+/-0.8 in euthyroid to 14.36+/-0.44 in hyperthyroid, and decreased in hypothyroid mitochondria to 8.62+/-0.63 nmol Ca(2+)/mg/s. Furthermore, the K(i) for the specific inhibitor Ru(360), depends on the thyroid status, i.e. 18, 19 and 13 nM for control, hyper- and hypothyroid mitochondria, respectively. In addition, the binding of 103Ru(360) was increased in hyperthyroid and decreased in hypothyroid mitochondria. Scatchard analysis for the binding of 103Ru(360) showed the following values: 28, 40 and 23 pmol/mg for control, hyper- and hypothyroid mitochondria, respectively. The K(d) for 103Ru(360) was found to be 30.39, 37.03 and 35.71 nM for controls, hyper- and hypothyroid groups, respectively. When hypothyroid rats were treated with thyroid hormone, mitochondrial Ca(2+) transport, as well as 103Ru(360) binding, reached similar values to those found for euthyroid mitochondria. PMID- 12781985 TI - Growth and hepatic acetyl coenzyme-A carboxylase activity are affected by dietary protein level in European seabass (Dicentrarchus labrax). AB - Two trials were undertaken with European seabass (Dicentrarchus labrax) to estimate the protein requirements for maintenance and growth as well as the effect of dietary protein level on the activity of hepatic acetyl coenzyme-A carboxylase (ACoAC). Six diets were formulated to contain graded levels of protein (from 5 to 55% crude protein (CP)) at a constant (12%) lipid level. Three other diets were also formulated to contain 35, 45 and 55% CP, but with a higher lipid level (19%). Groups of 10 individually marked fish (IBW: 100 g) and groups of 8 fish (IBW: 160 g) were used in trial I and II, respectively. Fish were fed to visual satiety and intake was recorded. At the end of both studies, whole body, liver and plasma samples were withdrawn for analyses. Growth rate was improved with increasing dietary CP level. Despite not being the object of a statistical analysis, feed efficiency tended to be enhanced at higher dietary CP level and protein efficiency ratio tended to decrease with increased protein intake. The reduction of the dietary protein/energy ratio, due to the increase of dietary lipids further improved growth and feed utilisation. Data from both experiments indicate 4.5+/-0.5 g kg(-1) d(-1) as the daily protein intake for maximum N gain and 520+/-50 mg kg(-1) d(-1) as the maintenance needs for nitrogen balance. An increase of dietary CP level, up to 25%, increased ACoAC activity. A further increase in dietary CP level (35 to 55%) did not affect liver ACoAC activity. The increase in dietary lipid level depressed significantly liver ACoAC specific activity. PMID- 12781986 TI - Neural stem cells constitutively secrete neurotrophic factors and promote extensive host axonal growth after spinal cord injury. AB - Neural stem cells (NSCs) offer the potential to replace lost tissue after nervous system injury. This study investigated whether grafts of NSCs (mouse clone C17.2) could also specifically support host axonal regeneration after spinal cord injury and sought to identify mechanisms underlying such growth. In vitro, prior to grafting, C17.2 NSCs were found for the first time to naturally constitutively secrete significant quantities of several neurotrophic factors by specific ELISA, including nerve growth factor, brain-derived neurotrophic factor, and glial cell line-derived neurotrophic factor. When grafted to cystic dorsal column lesions in the cervical spinal cord of adult rats, C17.2 NSCs supported extensive growth of host axons of known sensitivity to these growth factors when examined 2 weeks later. Quantitative real-time RT-PCR confirmed that grafted stem cells expressed neurotrophic factor genes in vivo. In addition, NSCs were genetically modified to produce neurotrophin-3, which significantly expanded NSC effects on host axons. Notably, overexpression of one growth factor had a reciprocal effect on expression of another factor. Thus, stem cells can promote host neural repair in part by secreting growth factors, and their regeneration-promoting activities can be modified by gene delivery. PMID- 12781987 TI - Astrocyte-targeted expression of IL-6 protects the CNS against a focal brain injury. AB - The effect of CNS-targeted IL-6 gene expression has been thoroughly investigated in the otherwise nonperturbed brain but not following brain injury. Here we examined the impact of astrocyte-targeted IL-6 production in a traumatic brain injury (cryolesion) model using GFAP-IL6 transgenic mice. This study demonstrated that transgenic IL-6 production significantly increased wound healing following the cryolesion. Thus, at 20 days postlesion (dpl) the GFAP-IL6 mice showed almost complete wound healing compared to litter mate nontransgenic controls. It seems likely that a reduced inflammatory response in the long term could be responsible for this IL-6-related effect. Thus, while in the acute phase following cryolesion (1-6 dpl) the recruitment of macrophages and T lymphocytes was higher in GFAP-IL6 mice, at 10-20 dpl it was significantly reduced compared to controls. Reactive astrogliosis was also significantly increased up to but not including 20 dpl in the GFAP-IL6 mice. Oxidative stress as well as apoptotic cell death was significantly decreased throughout the time period studied in the GFAP-IL6 mice compared to controls. This could be linked to the altered inflammatory response as well as to the transgenic IL-6-induced increase of the antioxidant, neuroprotective proteins metallothionein-I + II. These results indicate that although in the brain the chronic astrocyte-targeted expression of IL-6 spontaneously induces an inflammatory response causing significant damage, during an acute neuropathological insult such as following traumatic injury, a clear neuroprotective role is evident. PMID- 12781989 TI - Expression of the POU domain transcription factor, Oct-6, is attenuated in the adult mouse telencephalon, but increased by neurotoxic damage. AB - Oct-6 is a POU III domain transcription factor expressed in embryonic stem cells, Schwann cells, and neuronal subpopulations during telencephalic development. Its role is unknown except in Schwann cells where it is thought to regulate myelin specific gene expression. Expression of Oct-6 was recently discovered in neurons in postmortem human schizophrenic brain while being undetectable in matched controls. This study of human tissue contrasted in a number of regards with earlier studies of rodent brain and questioned what we can consider to be normal adult expression of this gene. In this study, we have investigated Oct-6 expression in normal adult mice and in mice treated with neuractive compounds. We show that Oct-6 is widely expressed in young adults but that its expression subsequently becomes restricted to specific neuronal subpopulations. Contrary to earlier reports, however, this specific expression is transient and is eventually completely lost from telencephalic neurons. The OCT-6 protein, somewhat surprisingly, is found to be cytoplasmic as well as nuclear in certain neuronal subpopulations. Finally, we report that neurotoxic doses of anticonvulsants reactivate OCT-6 expression in adult mouse brain. PMID- 12781990 TI - Downregulation and increased turnover of beta-amyloid precursor protein in skeletal muscle cultures by neuregulin-1. AB - The beta-amyloid precursor protein (betaAPP) is found in skeletal muscle localized to the base of the postsynaptic folds of the neuromuscular junction; yet here, as well as in neurons, its function remains enigmatic. Here we report that the motor nerve-derived trophic factor neuregulin-1 (NRG1) regulates both steady-state betaAPP levels as well as the metabolism of the cell surface associated protein in cultured muscle cells. These two effects occur over two discernible time scales. At short times (minutes to hours), NRG1 increases the rate of internalization and apparent degradation of cell surface betaAPP while reducing the release of soluble APP to the medium. At longer times (hours to days), NRG1 causes a decrease in mRNA for betaAPP with a concomitant reduction in steady-state protein levels. These are novel findings for this trophic factor originally identified as inducing the expression of nicotinic acetylcholine receptors and other important synaptic proteins in skeletal muscle. They suggest that betaAPP may play a receptor or signal transduction role at the neuromuscular junction since other receptor protein's actions are terminated in a similar fashion. The effects of NRG1 on betaAPP metabolism are overcome by inhibitors of both the phosphatidylinositol-3 (PI3) kinase and mitogen-activated protein (MAP) kinase pathways, yet are distinct from those activated during induction of nicotinic acetylcholine receptor biosynthesis. BetaAPP should be added to the list of specialized post-neuromuscular junction proteins that are regulated by cholinergic terminal-derived factors critical to synaptogenesis. PMID- 12781988 TI - Increased adenine nucleotide translocator 1 in reactive astrocytes facilitates glutamate transport. AB - A hallmark of central nervous system (CNS) pathology is reactive astrocyte production of the chronic glial scar that is inhibitory to neuronal regeneration. The reactive astrocyte response is complex; these cells also produce neurotrophic factors and are responsible for removal of extracellular glutamate, the excitatory neurotransmitter that rises to neurotoxic levels in injury and disease. To identify genes expressed by reactive astrocytes, we employed an in vivo model of the glial scar and differential display PCR and found an increase in the level of Ant1, a mitochondrial ATP/ADP exchanger that facilitates the flux of ATP out of the mitochondria. Ant1 expression in reactive astrocytes is regulated by transforming growth factor-beta1, a pluripotent CNS injury-induced cytokine. The significance of increased Ant1 is evident from the observation that glutamate uptake is significantly decreased in astrocytes from Ant1 null mutant mice while a specific Ant inhibitor reduces glutamate uptake in wild-type astrocytes. Thus, the astrocytic response to CNS injury includes an apparent increase in energy mobilization capacity by Ant1 that contributes to neuroprotective, energy-dependent glutamate uptake. PMID- 12781991 TI - FKBP12 mRNA expression is upregulated by intrinsic CNS neurons regenerating axons into peripheral nerve grafts in the brain. AB - We have examined the expression of the immunophilin FKBP12 in adult rat intrinsic CNS neurons stimulated to regenerate axons by the implantation of segments of autologous tibial nerve into the thalamus or cerebellum. After survival times of 3 days to 6 weeks, the brains were fresh-frozen. In some animals the regenerating neurons were retrogradely labelled with cholera toxin subunit B 1 day before they were killed. Sections through the thalamus or cerebellum were used for in situ hybridization with digoxygenin-labelled riboprobes for FKBP12 or immunohistochemistry to detect cholera toxin subunit B-labelled neurons. FKBP12 was constitutively expressed by many neurons, and was very strongly expressed in the hippocampus and by Purkinje cells. Regenerating neurons were found in the thalamic reticular nucleus and deep cerebellar nuclei of animals that received living grafts. Neurons in these nuclei upregulated FKBP12 mRNA; such neurons were most numerous at 3 days post grafting but were most strongly labelled at 2 weeks post grafting. Regenerating neurons identified by retrograde labelling were found to have upregulated FKBP12 mRNA. No upregulation was seen in neurons in animals that received freeze-killed grafts, which do not support axonal regeneration. We conclude that FKBP12 is a regeneration-associated gene in intrinsic CNS neurons. PMID- 12781992 TI - Inhibition of caspases promotes long-term survival and reinnervation by axotomized spinal motoneurons of denervated muscle in newborn rats. AB - We examined whether (1) a pan-caspase inhibitor, Boc-D-FMK, exerts long-term neuroprotective effects on spinal motoneurons (MNs) after root avulsion in neonatal rats and (2) whether the rescued spinal MNs regenerate their axons into a peripheral nerve (PN) graft and reinnervate a previously denervated target muscle. Eight weeks after root avulsion, 67% of spinal MNs remained in the Boc-D FMK-treated group, whereas all MNs died in the sham control group. By 12 weeks postinjury, however, all Boc-D-FMK treated MNs died. In the regeneration experiment, a PN graft was implanted at different times after injury. The animals were allowed to survive for 4 weeks following the operation. Without caspase inhibition, MNs did not regenerate at any time point. In animals treated with Ac DEVD-CHO, a caspase-3-specific inhibitor, and Boc-D-FMK, 44 and 62% of MNs, respectively, were found to regenerate their axons into a PN graft implanted immediately after root avulsion. When the PN graft was implanted 2 weeks after injury, however, MNs failed to regenerate following Ac-DEVD-CHO treatment, whereas 53% of MNs regenerated their axons into the graft after treatment with Boc-D-FMK. No regeneration was observed when a PN graft was implanted later than 2 weeks after injury. In the reinnervation study, injured MNs and the target biceps muscle were reconnected by a PN bridge implanted 2 weeks after root avulsion with administration of Boc-D-FMK. Eight weeks following the operation, 39% of MNs reinnervated the biceps muscle. Morphologically normal synapses and motor endplates were reformed in the muscle fibers. Collectively, these data provide evidence that injured neonatal motoneurons can survive and reinnervate peripheral muscle targets following inhibition of caspases. PMID- 12781993 TI - Repair of peripheral nerve transections with fibrin sealant containing neurotrophic factors. AB - Peripheral nerve injury is often followed by incomplete recovery of function and sometimes associated with neuropathic pain. There is, therefore, need for therapies which improve the speed of recovery and the final functional outcome after peripheral nerve injuries. In addition, neuropathic pain is not easily dealt with clinically and should preferably be eliminated. Neurotrophic factors have well-documented abilities to support neuron survival and stimulate neurite outgrowth, making them excellent candidates for use in repairing injured nerves. We investigated the possible beneficial effects of repairing the transected rat sciatic nerve by local application of a fibrin sealant containing nerve growth factor (NGF), glial cell line-derived neurotrophic factor (GDNF), or acidic fibroblast growth factor (aFGF). Fibrin sealant was used in conjunction with sutures. Evaluation of motor and sensory function, autotomy, and histological parameters was carried out from 1 to 12 weeks after injury. We demonstrate that NGF cotreatment decreased the occurance of autotomy, suggesting a reduction of neuropathic pain, and improved the performance in motor and sensory tests. In addition, the number of regenerating motoneurons was significantly increased after NGF administration. GDNF increased the speed of sensory recovery, but also markedly increased autotomy, indicating an increased degree of neuropathic pain. aFGF did not alter the outcome of the motor or sensory tests. Fibrin sealant could easily be used in conjunction with sutures to deliver neurotrophic substances locally to the damaged nerve and to enhance recovery of nerve function. PMID- 12781994 TI - Structural and functional neuroprotection in a rat model of Huntington's disease by viral gene transfer of GDNF. AB - Huntington's disease (HD) is an autosomal dominant disorder caused by an expanded polyglutamine (CAG) tract at the IT15 locus on chromosome 4. These excessive repeats lead to the degeneration of striatal and cortical neurons resulting in a devastating cognitive, psychiatric, and motor disorder for which no treatments are available. Neurotrophic factors support the viability of striatal neurons suggesting that they might prevent the inevitable neural degeneration and its accompanying functional decline associated with HD. The present study investigated whether glial cell line-derived neurotrophic factor (GDNF) delivered by an adeno associated virus could provide structural and functional neuroprotection in a rat model of HD. Lewis rats received bilateral injections of either AAV-GDNF (n = 12) or AAV-green fluorescence protein (AAV-GFP, n = 12) into the striatum followed 2 weeks later by chronic subcutaneous infusions of the mitochondrial toxin, 3-nitropropionic acid (3-NP, 38 mg/kg). All rats underwent 4 weeks of behavioral testing and were then sacrificed. Following 3-NP, the performance by AAV-GFP-treated rats on a raised platform motor task deteriorated while the performance by AAV-GDNF-treated rats was near normal (P < 0.001). AAV GDNF-treated rats also received better scores on a blinded semi-quantitative neurological scale compared to rats receiving AAV-GFP (P < 0.001). Histological analyses supported our behavioral findings. 3-NP-treated rats receiving AAV-GDNF displayed 70% more NeuN-immunoreactive neurons compared to 3-NP-treated rats receiving AAV-GFP (P = 0.002). Similar findings were seen with dopamine-and adenosine-3'5'-monophosphate-regulated phosphoprotein (DARPP-32) staining. These data indicate that the viral-mediated gene transfer of GDNF into the striatum provides neuroanatomical and behavioral protection in a rodent model of HD. PMID- 12781995 TI - Suppression of limbic motor seizures by electrical stimulation in thalamic reticular nucleus. AB - Kindling is a model of temporal lobe epilepsy in which repeated electrical stimulations in limbic areas lead to progressive increase of seizure susceptibility, culminating in generalized convulsions and the establishment of a permanent epileptic syndrome. We studied here the effect of stimulations in the thalamic reticular nucleus (TRN) on the development of seizures and hippocampal hyperexcitability in kindling elicited from the ventral hippocampus in rats. Animals given 12 kindling stimulations per day with 30-min intervals for 4 consecutive days developed generalized convulsions on day 4. Stimulations in TRN delivered simultaneously with those in the hippocampus induced marked suppression of seizure generalization. Similarly, the number of generalized seizures and the duration of behavioral convulsions were reduced when rats subjected to 40 kindling stimulations with 5-min intervals during about 3 h were costimulated in the TRN. The anticonvulsant effect of TRN costimulation was detected also when rats were test-stimulated in the hippocampus at 24 h and 2 and 4 weeks after the initial 40 hippocampal stimulations. Our data provide the first evidence that TRN stimulations can act to suppress limbic motor seizures in hippocampal kindling and suggest a new approach for seizure control in temporal lobe epilepsy. PMID- 12781996 TI - Quantitative analysis of perinatal rodent oligodendrocyte lineage progression and its correlation with human. AB - The development of a rodent model in the perinatal rat or mouse that reproduces the principal features of human perinatal white matter injury (periventricular leukomalacia) has been hampered by uncertainty about the developmental window in the rodent that coincides temporally with cerebral white matter development in the premature infant. We recently determined oligodendrocyte (OL) lineage progression in human cerebral white matter and found that the late OL progenitor (preOL) predominates throughout the high-risk period for periventricular leukomalacia [J. Neurosci. 21(2001), 1302-1312]. Here, we determined in the perinatal rat and mouse when each species displays a distribution of OL stages that is similar to the premature human cerebral white matter. PreOLs are abundant in the rat and mouse at P2. By P7, extensive OL maturation occurs in both species and coincides with the onset of early myelination. PreOLs and immature OLs mature in the P2 white matter along a medial to lateral gradient. This may provide an explanation for regional variation in the susceptibility of perinatal white matter to injury. We propose that the sequence of OL lineage progression is a useful means to estimate developmental windows of white matter maturation in perinatal rodents that coincide with those of developing human cerebral white matter. These studies support that the vulnerable period for white matter injury in the rodent is centered around P2 and should decline thereafter, coincident with the onset of myelination. PMID- 12781997 TI - Inhibition of endogenous VEGF impedes revascularization and astroglial proliferation: roles for VEGF in brain repair. AB - Vascular endothelial growth factor (VEGF) is upregulated following injury to the CNS. Our previous work has shown that exogenous application of VEGF promotes angiogenesis, blood-brain barrier permeability, and astroglial mitogenicity in the traumatized brain. To develop a model that could link endogenously secreted VEGF to brain tissue repair, a specific neutralizing antibody to VEGF was infused by osmotic minipump directly into the neocortex and striatum for up to 1 week. Tissues adjacent to the infusion/wound site were analyzed for specific vascular and astroglial protein markers and proliferation, necrosis/apoptosis (via TUNEL staining), VEGF, the VEGF receptors flt-1 and flk-1, and bFGF expression using immunohistochemistry and semi-quantitative RT-PCR. Neutralization of native VEGF caused significant decreases in angiogenic activity, astroglial proliferation, and nestin immunoexpression, while vascular and astroglial degeneration was substantially increased, resulting in much larger wound cavities when compared to controls. The hindrance of brain tissue repair occurred despite an increase in bFGF expression at the wound sites. VEGF appears to be an integral factor in CNS wound healing that is essential for vascular endothelial proliferation and survival and may also be necessary for astroglial proliferation and maintenance during the repair of brain injury. PMID- 12781998 TI - A new model for prenatal brain damage. I. GABAA receptor activation induces cell death in developing rat hippocampus. AB - Premature infants are at exceptionally high risk for hypoxic-ischemic insults and other traumatic events that result in permanent brain damage. However, no current models adequately mimic these events. An emerging concept is that the major excitatory drive in immature neurons is derived from depolarizing responses following activation of the gamma-aminobutyric acid (GABA)(A) receptor, resulting in the opening of voltage-sensitive calcium channels. While calcium-mediated signal transduction is trophic in developing neurons, excessive calcium entry is a major mediator of excitotoxicity. We report that exogenous activation of GABA(A) receptors by muscimol in newborn rats increases cell death in the hippocampus. The effects are region specific, persistent, and greater in males. Muscimol-induced damage is prevented by pretreatment with diltiazem, an L-type voltage-sensitive calcium channel blocker. Results using hippocampal cultures parallel those observed in vivo, indicating that the effects are mediated directly in the hippocampus. Existing models of pediatric hypoxic-ischemic brain damage focus on the effects of glutamate in the postnatal day 7 rat, because it is considered analogous to the newborn human. This makes the newborn rat analogous to the late gestational human. Ischemia in newborn rats induces GABA release and we propose that treatment with muscimol mimics the cell death cascade induced by hypoxia-ischemia in premature human infants. PMID- 12781999 TI - A novel model for prenatal brain damage. II. Long-term deficits in hippocampal cell number and hippocampal-dependent behavior following neonatal GABAA receptor activation. AB - Premature infants are at especially high risk for asphyxia, seizures, and other conditions that cause hypoxia-ischemia. These events result in abnormal brain pathology and behavioral deficits that persist throughout adolescence and into adulthood. Current rodent models of human infant hypoxic-ischemic brain damage have focused on exogenous glutamate receptor agonist exposure in the postnatal day 7 rat. While this model is considered analogous to the newborn human, no adequate models for preterm infant brain damage have been developed. Recent work from our lab has proposed a potential model for preterm infant brain damage in which neonatal rats are treated with exogenous muscimol, the selective gamma aminobutyric acid(A) (GABA(A)) receptor agonist, on postnatal days 0 and 1. In the companion paper to this one (Exp. Neurol., in press), we report fewer neurons in the hippocampal formation on postnatal day 7 (6 days after treatment), but the persistence of these anatomical deficits, and potential resultant behavioral dysfunctions, were not investigated. In the current experiment, we documented that muscimol exposure on postnatal days 0 and 1 leads to fewer neurons in the male and female rat hippocampus (CA1, CA2/3, and dentate gyrus) on postnatal day 21. Also, neonatal muscimol exposed males and females displayed deficits on hippocampal-dependent learning tasks such as a preweanling version of the Morris water maze task and the open field task. We conclude that exposure to exogenous GABA(A) receptor activation over the first 2 days of postnatal life, a model for preterm infant hypoxic injury, produces anatomical and behavioral deficits observed into adolescence. PMID- 12782000 TI - Involvement of apoptosis and calcium mobilization in tetrahydrobiopterin-induced dopaminergic cell death. AB - Parkinson's disease is a neurodegenerative disorder associated with selective loss of the dopaminergic neurons in the substantia nigra. We have previously shown that tetrahydrobiopterin (BH4), the obligatory cofactor for dopamine synthesis, exerts selective toxicity on dopamine-producing cells. In the present study we determined, both in vitro and in vivo, whether the cell death induced by this endogenous molecule involves apoptosis, resembling that which occurs in Parkinson's disease. Transmission electron microscopic analysis revealed that the dopamine-producing CATH.a cells underwent ultrastructural changes typical of apoptosis, such as cell shrinkage and chromatin condensation, upon exposure to BH4. The BH4 treatment also caused intranuclear DNA fragmentation as determined by TUNEL staining. A similar phenomenon also occurred in vivo, as the nigral cells became TUNEL-positive upon injection of BH4 into the substantia nigra. The BH4-induced CATH.a cell death seemed to involve macromolecule synthesis because cycloheximide and actinomycin D had protective effects. Concurrent treatment with the caspase inhibitor Z-VAD-FMK also suppressed cell death. BH4 treatment led to increases in the ratio of Bax/Bcl-x(L) mRNA and protein levels. Ca(2+) seemed to play a role in BH4-induced cell death, because BH4 caused an increase in Ca(2+) uptake and the intracellular Ca(2+) release blocker dantrolene, intracellular Ca(2+) chelator BAPTA/AM, and extracellular Ca(2+) chelator EGTA each attenuated the toxicity. These data provide evidence that the dopaminergic cell death induced by BH4 involves apoptosis and suggest relevance of this cell death to degeneration of the dopaminergic system in Parkinson's disease. PMID- 12782001 TI - Hypothermic preconditioning induces rapid tolerance to focal ischemic injury in the rat. AB - Stressful, preconditioning stimuli can elicit rapid and delayed forms of tolerance to ischemic injury. The identification and characterization of preconditioning stimuli that are effective, but relatively benign, could enhance the clinical applicability of induced tolerance. This study examines the efficacy of brief hypothermia as a preconditioning stimulus for inducing rapid tolerance. Rats were administered hypothermic preconditioning or sham preconditioning and after an interval of 20-120 min were subjected to transient focal ischemia using a three-vessel occlusion model. The volume of cerebral infarction was measured 24 h or 7 days after ischemia. In other experiments, the depth or duration of the hypothermic stimulus was manipulated, or a protein synthesis inhibitor (anisomycin) was administered. Twenty minutes of hypothermia delivered 20 or 60 (but not 120) min prior to ischemia significantly reduces cerebral infarction. The magnitude of protection is enhanced with deeper levels of hypothermia, but is not affected by increasing the duration of the hypothermic stimulus. Treatment with a protein synthesis inhibitor does not block the induction of rapid tolerance. Hypothermic preconditioning elicits a rapid form of tolerance to focal ischemic injury. Unlike delayed tolerance induced by hypothermia, rapid tolerance is not dependent on either de novo protein synthesis or the duration of the preconditioning stimulus. These findings suggest that the mechanisms underlying rapid and delayed tolerance induced by hypothermia differ fundamentally. Brief hypothermia could provide a rapid means of inducing transient tissue protection in the context of predictable ischemic events. PMID- 12782002 TI - Testosterone, but not nonaromatizable dihydrotestosterone, improves working memory and alters nerve growth factor levels in aged male rats. AB - Recent studies have suggested that testosterone levels are lower in men with Alzheimer's disease and that testosterone treatment improves cognition in older men. Since testosterone can be aromatized to estrogen, testosterone's effects could be due to conversion into estrogen. We treated aged male rats with either testosterone or dihydrotestosterone (DHT), the latter of which is not aromatized to estrogen, in order to determine whether these treatments improve spatial working and reference memory as assessed in the water radial arm maze. We also tested whether such effects are related to beta-amyloid levels in the hippocampus or neurotrophin levels in the hippocampus, entorhinal cortex, frontal cortex, or striatum. Aged rats made more errors than young rats on all memory measures. Testosterone, but not DHT, improved working memory and decreased hippocampal NGF protein in aged rats, while having no effect on beta-amyloid. However, higher beta-amyloid levels were correlated with poorer working memory performance in young rats. Neurotrophin levels in entorhinal cortex were positively correlated with errors for all memory measures in androgen-treated rats. Similar to findings in human studies, in our study androgen treatment lowered circulating estradiol levels in aged rats, suggesting that androgen treatment exerts feedback to the hypothalamic pituitary axis and that conversion to estrogen may not be the underlying biological mechanism of testosterone's effects on memory and growth factor levels. The ratio of estradiol to testosterone, or the actions of the aromatase enzyme itself, may be responsible for the observed effects. These data support the hypothesis that testosterone therapy in aging men may provide positive effects on cognition and that neural regions that are linked to cognition, such as the hippocampus and/or entorhinal cortex, may be involved in such effects. PMID- 12782003 TI - Denervation-activated inward rectifier in frog slow skeletal muscle fibers. AB - We tested whether the absence of an inward rectifier channel in slow skeletal muscle fibers of the frog is regulated by innervation. Normal and denervated slow fibers were identified according to their passive electrical properties. In current-clamp experiments, anomalous rectification was quantified as the ratio of effective resistances for hyperpolarizing and depolarizing pulses. In isotonic potassium solution, this ratio was 0.45 +/- 0.1 (n = 14) for twitch fibers, whereas slow fibers displayed linear behavior [ratio = 1.0 +/- 0.05 (n = 15)]. However, denervated slow fibers showed anomalous rectification (ratio, 0.48 +/- 0.07; n = 5). This finding was supported by voltage-clamp experiments in which denervated slow fibers displayed (1) an inward rectifier current during hyperpolarizing pulses, (2) an increase in this current when [K(+)](o) was increased, and (3) a current inhibition after application of Ba(2+). These results suggest that frog slow fibers, which normally do not possess inward rectifier channels, can express them after denervation. PMID- 12782004 TI - Neuronal loss in familial frontotemporal dementia with ubiquitin-positive, tau negative inclusions. AB - The neuronal density in the frontal, temporal, and parietal lobes was determined in nine cases of familial frontotemporal dementia with ubiquitin-positive, tau negative inclusions (FTDU). The mean age at onset was 56.9 +/- 2.2 years and the duration of disease was 6.7 +/- 0.5 years. The mean age at death was 63.6 +/- 2.2 years. There was substantial loss (34%) of brain weight (877 +/- 73 g) in the familial cases in comparison with 10 normal aged controls (1326 +/- 50 g, P < 0.001). All of the familial FTDU cases showed atrophy of the frontal, temporal, and parietal lobes; neuronal loss; vacuolation in superficial laminae; reactive astrocytosis; and ubiquitin-positive, tau-negative intracytoplasmic and intranuclear inclusions and dystrophic neurites in varying sites and numbers. Neuronal loss was estimated in nine cases of familial FTDU and in 10 aged controls using a stereological probe, the optical "disector," and a computerized stereology system (CAST-Grid, Olympus, Denmark). There was a significant reduction in neuronal density in the frontal lobe (22.3 +/- 3.8 x 10(3)/mm(3)) of familial FTDU in comparison to aged controls (33.1 +/- 1.7 x 10(3) per mm(3), P < 0.05). An estimate of the relative numbers of neurons was calculated by multiplying the numerical density by the cortical thickness, which showed a striking loss of neurons of 56% in the frontal lobe, 52% loss in the temporal lobe, and a 49% loss in the parietal lobe of familial FTDU when compared to controls. This study shows that familial FTDU has profound focal neuronal loss in multiple association areas that relate to the clinical symptoms characteristic of the disease. PMID- 12782005 TI - Effects of seizure severity and seizure repetition on postictal cardiac arrhythmia following maximal electroshock. AB - Convulsive seizures triggered by maximal electroshock (MES) induce profound abnormalities in neural regulation of cardiac rhythm that are manifested by a period of marked cardiac arrhythmia in the immediate postictal state. It is not known whether seizure severity or seizure experience may influence the duration of cardiac arrhythmia in the postictal state. We varied the duration of MES administered to rats to vary seizure severity, as measured by the extensor to flexion (E/F) ratio. In separate experiments, rats were subjected to daily MES. Finally, we pretreated rats with ketamine prior to MES to block seizures hindlimb extension. In all animals, the R-R interval was plotted on the tachogram, and the duration of the arrhythmia was measured. Increases in MES duration increased significantly the E/F ratio and prolonged significantly the postictal cardiac arrhythmia. Repetition of MES caused a kindling effect with respect to seizure severity resulting in a significant increase of the E/F ratio and significant increases in the duration of postictal arrhythmia. Blocking of the hindlimb extension by ketamine abolished arrhythmia suggesting that the arrhythmia is not caused directly by MES. Severity of tonic convulsive seizures is a determinant of disordered cardiac autonomic regulation and directly influences the duration of cardiac arrhythmia during the immediate postictal state following MES. Seizure repetition also increases abnormalities of postictal neural regulation of the heart, but further studies are needed to determine whether this effect is independent of seizure severity increases. PMID- 12782006 TI - Complex intracellular inclusions in the brain of a child with a stealth virus encephalopathy. AB - Unusual pigmented intracellular inclusions are commonly seen in cultures obtained from patients infected with stealth viruses. Some of these structures may potentially provide a source of chemical energy for the infected cells to help compensate for the apparent damage to the cells' mitochondria. They have accordingly been termed alternative cellular energy pigments (ACE pigments). In keeping with this suggestion, the present paper illustrates the diversity of extraneous materials present in vacuolated, mitochondria-damaged cells seen in the brain biopsy of a child with a stealth-virus-associated encephalopathy. Many of the intracellular inclusions show highly ordered structuring, while others have a more amorphous appearance. These structures may provide a target for energy-based therapeutic intervention in stealth-virus-infected patients. PMID- 12782007 TI - Stealth virus culture pigments: a potential source of cellular energy. AB - The cytopathic effect (CPE) induced in human MRC-5 fibroblasts by blood mononuclear cells of patients infected with stealth-adapted viruses is characterized by the formation of clusters of foamy vacuolated cells that commonly become heavily pigmented. The pigmented material coalesces into discrete structures of varying shapes and sizes, including solid particles, flat ribbons, and long, thin threads. Pigmented material can also be observed in long-term culture supernatants, sometimes accompanied by solid and needle-shaped lipid-like crystals. Accumulation of the pigmented material correlates with lessening of the CPE and outgrowth of normal appearing cells from cell clusters. The CPE can be rapidly reactivated by replacing the culture supernatant with fresh culture medium. Conversely, reactivation can be partially prevented by the addition of particulate pigmented material to fresh culture medium. The pigmented material displays striking luminescence and autofluorescence. Occasional particles are ferromagnetic. Varying percentages of different minerals are identifiable in the particles using energy dispersive X-ray (EDX) analysis. Metabolic studies indicate the particles can have reducing (electron donating) capacity and can liberate gas. These findings, along with the extended survival of viable cells in unfed virus-infected cultures, suggest that the pigments are a potential source of cellular energy. The presence of light and magnetic-sensitive alternative cellular energy (ACE) pigments has opened new approaches for the potential therapy of stealth virus-infected patients. PMID- 12782008 TI - Alu profiling of primary and metastatic nonsmall cell lung cancer. AB - The metastatic potential of nonsmall cell carcinoma of lung (NSCLC), is currently recognized post factum, when lymph nodes or distant organs are already involved. Our ability to determine which tumors have acquired metastatic potential could help direct therapy to be more aggressive or less aggressive based upon this information. Evaluation of microsatellite instability via detection of LOH at specific loci may be useful in identifying specific markers and/or genes associated with this process. We examined Alu insertional elements as a potential marker of genetic changes associated with the metastatic potential of NSCLC. We analyzed archived, paraffin embedded tissue from 20 proven cases of NSCLC. DNA was extracted from 10 micron paraffin sections and amplified using an Alu PCR protocol. This technique does not examine specific loci but rather results in a banding profile of cellular genomic DNA. Informative allelic banding patterns, noted as differences between primary and metastatic lesions from the same patient, were observed in five of six cases (83%) with intrapulmonary metastases and in only nine of 14 (64%) cases with extrapulmonary metastases. Multiple genomic changes were detected in metastatic tumor cells as compared to normal lung tissue or primary lung tumor tissue. It appears that Alu profiling may be useful in the detection of metastatic vs primary lesions, and this technique may offer a method for identifying novel genes responsible for tumor progression and metastases. PMID- 12782009 TI - Vascular expression of matrix metalloproteinase-13 (collagenase-3) in basal cell carcinoma. AB - Matrix metalloproteinase-13 (MMP-13; collagenase-3) was detected in the vasculature from 17 of 20 human basal cell carcinomas as assessed by immunohistology immediately after surgery. In contrast, MMP-1 (interstitial collagenase) was detected in the vasculature of only two of the same specimens. MMP-13 reactivity was also observed in the capillaries of normal human skin taken from the wound margin. Human dermal microvascular endothelial cells as well as human umbilical vein endothelial cells were isolated in culture and examined for MMP-13 expression. By reverse transcription-polymerase chain reaction and Southern blotting, an MMP-13 transcript was detected in unstimulated endothelial cells. The transcript was upregulated in cells treated with 50 nM phorbol myristate acetate (PMA). Western blotting demonstrated the presence of an anti MMP-13 - immunoreactive protein in culture fluid from both cell sources. Immunoreactivity was stronger in culture fluid from cells treated with interleukin-1alpha (IL-1alpha) than in culture fluid from control cells. Tumor necrosis factor-alpha (TNF-alpha) and PMA also upregulated MMP-13 expression but these agents were not as effective as IL-1alpha. Additionally, reactivity was greater in culture fluid from endothelial cells grown on three-dimensional lattices of polymerized type I collagen than on dried collagen films. These data indicate that endothelial cells in the skin are a source of MMP-13 and that enzyme expression is upregulated under conditions that promote endothelial cell growth and vascular differentiation. PMID- 12782010 TI - Skeletal muscle myosin is the autoantigen for experimental autoimmune myositis. AB - Experimental autoimmune myositis (EAM) is a rodent model for human inflammatory muscle disease (IMD). It can be induced by immunization of rodents with skeletal muscle homogenate and adjuvant. The specific myositogenic autoantigen has not been clearly identified although some evidence points to skeletal muscle myosin. In this report we strengthen this evidence, showing that Lewis rats immunized with purified skeletal muscle myosin develop EAM with the same pattern and severity as EAM induced by whole rabbit skeletal muscle homogenate (WRM). Multiple inflammatory lesions are detected histopathologically in the biceps, quadriceps, and gastrocnemius muscles. Myosin-reactive T cells from animals immunized either with myosin or with WRM have similar patterns of antigen-induced proliferation. The results show that myosin, a component of skeletal muscle, is at least one autoantigen in EAM. PMID- 12782011 TI - Platelet-rich plasma gel promotes differentiation and regeneration during equine wound healing. AB - Nonhealing wounds of the lower equine limb represent a challenging model. The platelet is a natural source of a myriad of growth factors and cytokines that promote wound healing. This study evaluates the potential of platelet derived factors to enhance wound healing in the lower equine limb. Platelets were isolated from horse blood and activated with thrombin, a process known to induce growth factor release. This produced a platelet gel composed of platelet-rich plasma (PRP). To test this all-natural wound healant, 2.5-cm(2) full thickness cutaneous wounds were created below the knee and hock of a thoroughbred horse. Wounds were treated with PRP gel or left untreated. Sequential wound biopsies collected at Days 7, 36, and 79 postwounding permitted comparison of the temporal expression of differentiation markers and wound repair. To test the hypothesis that wounds treated with PRP gel exhibit more rapid epithelial differentiation and enhanced organization of dermal collagen compared to controls, tissues were stained for cytokeratin 10, a suprabasal differentiation marker, and the reestablishment of collagen was evaluated by trichrome staining. PRP gel-treated wounds at Day 7 expressed intense cytokeratin 10 staining near the wound junction in suprabasal epidermal layers, while staining in control tissues was less intense and restricted to apical epidermal layers distal to the wound junction. By Day 79, the staining was equal in both groups. However, PRP gel-treated wounds at Day 79 contained abundant, dense collagen bundles oriented parallel to each other and to the overlying epithelium, whereas control tissues contained fewer collagen fibers that were oriented randomly. Thus, treatment of wounds with PRP gel induced accelerated epithelial differentiation and produced tissue with organized, interlocking collagen bundles. This study reveals that this novel all natural wound healant induced wound repair in injuries previously deemed untreatable. PMID- 12782012 TI - Activation of a caspase-dependent oxidative damage response mediates TGFbeta1 apoptosis in rat hepatocytes. AB - Activation of transforming growth factor-beta type 1- (TGFbeta1) mediated signaling occurs in response to cell injury affecting stem-type cells and hepatocytes in liver. In this work we used WB stemlike liver epithelial cells and p53-defective CWSV-1 nontumorigenic rat hepatocytes to investigate the possible roles of caspases and oxidative stress in TGFbeta1 signaling. TGFbeta1 significantly increased the level of 4-hydroxy-2-nonenal (4-HNE), a stable product of lipid peroxidation. In addition, TGFbeta1-treated cells exhibited activation of caspases that accompanied by enhanced cleavage of the caspase substrate poly(ADP)-ribose polymerase (PARP) and induction of apoptosis. WB cells were twice as sensitive as sensitive as CWSV-1 cells to induction of TGFbeta1 apoptosis. TGFbeta1-apoptosis was significantly reduced when cells were treated with TGFbeta1 in the presence of inhibitors of caspase-1, -3, -8, and -9. Importantly, in addition to suppression of apoptosis, treatment of cells with the caspase-3 inhibitor Z-DEVD-FMK in the presence of TGFbeta1 suppressed the formation 4-HNE and restored mitotic activity. Together, these data suggest TGFbeta1 induces activation of a caspase signaling cascade that includes an oxidative damage response, PARP cleavage, and apoptosis that do not require intact p53 in rat hepatocytes. PMID- 12782013 TI - Downregulation of CD44v6 in colorectal carcinomas is associated with hypermethylation of the CD44 promoter region. AB - Overexpression of the cell adhesion protein CD44v6 has been demonstrated in colorectal cancer and other gastrointestinal tumors. While CD44v6 is upregulated in benign colorectal adenomas and well-differentiated colorectal cancer tissues, downregulation frequently occurs during disease progression. The mechanism of downregulation, however, is unknown. Therefore, we evaluated the methylation status of the CD44 promoter as a mechanism for decreased CD44v6 expression in advanced colorectal carcinomas. We demonstrated by methylation-sensitive restriction enzyme digestion that the CpG islands of the CD44 promoter were methylated in 6/21 (28%) of benign colorectal adenomas. Interestingly, in colorectal carcinomas the frequency of promoter methylation was significantly increased (10/19; 53%) compared to 7/21 (33%) in the corresponding normal mucosa. Methylation seems to be associated with a more advanced cancer stage, but the trend did not reach statistical significance. In colorectal carcinomas with CD44 promoter methylation CD44v6 mRNA was detected by reverse transcription-polymerase chain reaction in 3/10 carcinomas, whereas in tumors without CD44 promoter methylation CD44v6 expression was observed in 8/9 (P 50 cyclotides. To facilitate their sequencing by MS MS, an analytical strategy is presented involving aminoethylation of cysteines. This overcomes a number of problems intimately associated with the cyclotide core structure-that is, their joined N and C termini, disulfide knot, and low or clustered content of positively charged amino acids and enzymatic cleavage sites. As a result, charges as well as cleavage sites are introduced at the most conserved part of their sequence, the cysteines. Combined with tryptic digestion, all intercysteine loops are then of suitable size and charge for MS-MS sequencing. The utility of this strategy is shown by the sequencing of two novel cyclotides isolated from V. cotyledon; vico A (cyclo (AESCVYIPCFTGIAGCSCKNKVCYYNGSIPC)) and vico B (cyclo (AESCVYIPCITGIAGCSCKNKVCYYNGSIPC)); their complete sequence could be determined by nanospray MS-MS. The strategy for converting conserved cysteines to enzymatic cleavage sites might also benefit the study of other peptides and proteins displaying similar structural problems for MS analysis. PMID- 12782039 TI - The changes in crosslink contents in tissues after formalin fixation. AB - The aim of this study was to detect crosslinks of collagen and elastin in formalin-fixed tissue, to perform quantification of these crosslinks, and to investigate the effects of formalin fixation on crosslink contents in human yellow ligament and cartilage. Pyridinoline (Pyr) is a stable and nonreducible crosslink of collagen. Pentosidine (Pen) is a senescent crosslink formed between arginine and lysine in matrix proteins, including collagen. Desmosine (Des) and its isomer isodesmosine (Isodes) are crosslinks specifically found in elastin. It is useful to measure crosslink contents of collagen and elastin as a way of investigating the properties of various tissues or their pathological changes. If it is possible to evaluate crosslinks of collagen and elastin in formalin-fixed tissues, we can investigate crosslinks in a wide variety of tissues. We used HPLC to compare the concentrations of Pyr, Pen, Des, and Isodes in the formalin-fixed tissues with their concentrations in the frozen tissues. Pyr and Pen were detected in both the formalin-fixed yellow ligament and the cartilage, and their concentrations were not significantly affected by or related to the duration of formalin fixation. Des and Isodes were detected in the formalin-fixed yellow ligament but in significantly lower amounts compared to the frozen samples. We concluded that crosslinks of collagen were preserved in formalin, but crosslinks of elastin were not preserved in it. The reason for this might be that formalin did not fix elastin tissues sufficiently or it destroyed, masked, or altered elastin crosslinks. PMID- 12782040 TI - Poly(A) tailing of ancient DNA: a method for reproducible microsatellite genotyping. AB - Microsatellites could be of great potential use in the analysis of ancient remains, but so far such analyses have failed to be reproducible mainly because of the high degree of ancient DNA (aDNA) degradation. During PCR, annealing of the primers to the complementary sequences of microsatellites occurs together with cross-annealing of partially degraded repeated sequences. This could create chimeric alleles that do not correspond to the authentic ones. Here we report a simple method for processing aDNA fragments prior to PCR that greatly reduces the production of chimeric alleles. This approach eliminates aDNA molecules broken within the repeats as targets for Taq polymerase by adding poly(A) tails at the 3(') ends of the DNA fragments, which disrupts the homology in the region and thus prevents annealing out of register. We have analyzed one dinucleotide- (D6S337) and two trinucleotide-containing loci (IT15 and SCA1) using poly(A) tailed and the same untreated aDNA as template. aDNAs were isolated from 28 human remains, 600 and 7000 years of age. In repeated experiments with untreated aDNAs we obtained three to five times more alleles compared to poly(A)-tailed aDNAs. According to our results, modification of aDNA by poly(A) tailing is an efficient pretreatment for accurate genotyping. PMID- 12782041 TI - Superoxide dismutases from the oyster parasite Perkinsus marinus: purification, biochemical characterization, and development of a plate microassay for activity. AB - We have isolated and biochemically characterized superoxide dismutase (SOD) activity in cell extracts of clonally cultured Perkinsus marinus, a facultative intracellular parasite of the Eastern oyster, Crassostrea virginica. In order to assess the SOD activity throughout the purification, we developed and optimized a 96-well-plate microassay based on the inhibition of pyrogallol oxidation. The assay was also adapted to identify SOD activity type (Cu/Zn-, Mn-, or FeSOD), even in mixtures of more than one type of SOD. All SOD activity detected in the cell extracts was of the FeSOD type. Most of the SOD activity in P. marinus trophozoites resides in a major component of subunit molecular weight 24 kDa. The protein was purified by affinity chromatography on an anti-SOD antibody-Sepharose column. Amino-terminal peptide sequence of the affinity-purified protein corresponds to the predicted product of the PmSOD1 gene and indicates that amino terminal processing has taken place. The results are discussed in the context of processing of mitochondrially targeted SODs. PMID- 12782042 TI - Characterization of proton abstraction steps in enzymatic reactions by Fourier transform infrared spectroscopy. PMID- 12782043 TI - An improved synthetic and purification procedure for the hydrophobic segment of the transmembrane peptide phospholamban. PMID- 12782044 TI - Cytotoxic effect of doxycycline and its implications for tet-on gene expression systems. PMID- 12782045 TI - Automated sample preparation using vapor-phase hydrolysis for amino acid analysis. PMID- 12782047 TI - Technical and regulatory hurdles for DNA vaccines. AB - DNA vaccines have been widely used in laboratory animals and non-human primates over the last decade to induce antibody and cellular immune responses. This approach has shown some promise, in models of infectious diseases of both bacterial and viral origin as well as in tumour models. Clinical trials have shown that DNA vaccines appear safe and well tolerated, but need to be made much more potent to be candidates for preventive immunisation of humans. This review describes recent work to improve the delivery of plasmid DNA vaccines and also to increase the immunogenicity of antigens expressed from the DNA vaccine plasmids, including various formulations and molecular adjuvants. Because DNA vaccines are relatively new and represent a novel vaccine technology, certain safety issues, such as the potential for induction of autoimmune disease and integration into the host genome, must be examined carefully. If potency can be improved and safety established, plasmid DNA vaccines offer advantages in speed, simplicity, and breadth of immune response that may be useful for the immunisation of humans against infectious diseases and cancers. PMID- 12782048 TI - Recent advances in veterinary vaccine adjuvants. AB - Next generation veterinary vaccines are going to mainly comprise of either subunit or inactivated bacteria/viruses. These vaccines would require optimal adjuvants and delivery systems to accord long-term protection from infectious diseases in animals. There is an urgent need for the development of new and improved veterinary and human vaccine adjuvants. Adjuvants can be broadly divided into two classes, based on their principal mechanisms of action: vaccine delivery systems and 'immunostimulatory adjuvants'. Vaccine delivery systems are generally particulate e.g. emulsions, microparticles, ISCOMS and liposomes, and mainly function to target associated antigens into antigen presenting cells (APC). In contrast, immunostimulatory adjuvants are predominantly derived from pathogens and often represent pathogen associated molecular patterns, e.g. LPS, MPL and CpG DNA, which activate cells of the innate immune system. Recent progress in innate immunity is beginning to yield insight into the initiation of immune responses and the ways in which immunostimulatory adjuvants might enhance this process in animals and humans alike. PMID- 12782049 TI - Plant-based vaccines. AB - Plant systems are reviewed with regard to their ability to express and produce subunit vaccines. Examples of different types of expression systems producing a variety of vaccine candidates are illustrated. Many of these subunit vaccines have been purified and shown to elicit an immune response when injected into animal models. This review also includes vaccines that have been administered orally in a non-purified form as a food or feed product. Cases are highlighted which demonstrate that orally delivered plant-based vaccines can elicit immune responses and in some case studies, confer protection. Examples are used to illustrate some of the inherent advantages of a plant-based system, such as cost, ease of scale-up and convenience of delivery. Also, some of the key steps are identified that will be necessary to bring these new vaccines to the market. PMID- 12782050 TI - Haemolysin A and listeriolysin--two vaccine delivery tools for the induction of cell-mediated immunity. AB - Haemolysin A of Escherichia coli and listeriolysin of Listeria monocytogenes represent important bacterial virulence factors. While such cytolysins are usually the reason for morbidity and even mortality, vaccine researchers have turned haemolysin A and listeriolysin into tools for vaccine delivery. Both cytolysins have found widespread application in vaccine research and are highly suitable for the elicitation of cell-mediated immunity. In this paper, we will review vaccine delivery mediated by the haemolysin A secretion system and listeriolysin and will highlight their use in vaccination approaches against protozoan parasites. PMID- 12782051 TI - A review of the effectiveness of vaccine potency control testing. AB - The use of potency control testing is a valuable tool for testing the actual relative strength of manufactured assembly lots of vaccine. Biological-based manufacturing methods are inherently variable and potency testing is a tool to ensure lot-to-lot consistency of commercial vaccines. A strong historical link to clinical efficacy has been established where correlation to efficacy and adequate test validation have been achieved. The link to immunogenicity and efficacy has traditionally been strongest with attenuated vaccines and toxoids. Control potency test failure does predict that a serial or batch of vaccine would most likely provide insufficient immunogenicity in typical field applications. Because of the complexity of pathogenic processes and associated immune responses, potency tests may not always directly predict the effectiveness of a vaccine. Thus, vaccines that pass control potency testing may not always provide adequate efficacy. This is particularly true of adjuvanted, inactivated vaccines. In the development of vaccine formulations and control tests for vaccines, the nature of the desired protective immune responses to the targeted pathogen (when known) should be considered. These considerations could provide better alternatives in the assays chosen as correlates of immunity and may more accurately predict efficacy and assure batch-to-batch consistency. Also, the effects of the dose and duration of antigen exposure as well as the nature of antigen presentation and generation of extrinsic cytokines could be characterised and correlated to vaccine potency as additional indicators of vaccine efficacy. PMID- 12782052 TI - Hepatitis vaccines: recent advances. AB - Despite the availability of hepatitis A vaccines that might provide protection for decades, hepatitis B vaccines that provides protection for at least 15 years and the recent introduction of a combined hepatitis A and B vaccine, these infections continue to spread in both the developed and developing world. Hepatitis A vaccine coverage has been limited to high-risk groups: such a selective immunisation policy is unlikely to have a major impact. If adequate immunogenicity in infants is confirmed, dosing schedules can be improved and the costs of vaccination reduced, universal paediatric immunisation with combined hepatitis A and B products is likely to result in the eventual eradication of these infections. In the interim, novel hepatitis A vaccines are being investigated and additional studies on hepatitis A vaccine immunogenicity in infants are in progress. Worldwide use of hepatitis B vaccines for the newborn, young children and high-risk groups should control this infection and obviate the need for a vaccine against hepatitis D. Newer hepatitis B vaccines that may reduce the likelihood of non-responsiveness and have immunotherapeutic value are under study. A recombinant hepatitis E vaccine for use in endemic regions is currently in clinical trials. The development of an effective hepatitis C vaccine has been agonisingly slow and many impediments have been recognised. These include the lack of a susceptible small animal, a high degree of hepatitis C virus (HCV) genomic diversity and failure to produce high quantities of HCV in tissue culture. The development of a novel HCV replicon system may be a major breakthrough. Nonetheless, it may still be exceedingly difficult to produce a vaccine that uniformly provides sterilising immunity; the possibility of developing a hepatitis C vaccine that can prevent chronic infection is an exciting concept that requires further investigation. Advances in recombinant technology, the use of novel genetic (DNA-based) vaccines, expression of hepatitis antigens in plants and improved adjuvants also hold considerable promise. PMID- 12782053 TI - Experimental vaccines against measles in a world of changing epidemiology. AB - Vaccination with the current live attenuated measles vaccine is one of the most successful and cost-effective medical interventions. However, as a result of persisting maternal antibodies and immaturity of the infant immune system, this vaccine is poorly immunogenic in children <9 months old. Immunity against the live vaccine is less robust than natural immunity and protection less durable. There may also be some concern about (vaccine) virus spread during the final stage of an eventual measles eradication program. Opinions may differ with respect to the potential threat that some of these concerns may be to the World Health Organisation goal of measles elimination, but there is a consensus that the development of new measles vaccines cannot wait. Candidate vaccines are based on viral or bacterial vectors expressing recombinant viral proteins, naked DNA, immune stimulating complexes or synthetic peptides mimicking neutralising epitopes. While some of these candidate vaccines have proven their efficacy in monkey studies, aerosol formulated live attenuated measles vaccine are evaluated in clinical trials. PMID- 12782054 TI - Recent progress in the development and testing of vaccines against human tuberculosis. AB - The growing pandemic of human tuberculosis has not been affected significantly by the widespread use of the only currently available vaccine, bacille Calmette Guerin. Bacille Calmette Guerin protects uniformly against serious paediatric forms of tuberculosis and against adult pulmonary tuberculosis in some parts of the world, but there are clearly populations in high-burden countries which do not benefit from the current vaccination regimen. New tuberculosis vaccines will be essential for the ultimate control of this ancient disease. Research over the past 10 years has produced literally hundreds of new tuberculosis vaccine candidates representing all of the major vaccine design strategies; protein/peptide vaccines in adjuvants, DNA vaccines, naturally and rationally attenuated strains of mycobacteria, recombinant mycobacteria and other living vaccine vectors expressing genes coding for immunodominant mycobacterial antigens, and non-peptide vaccines. Many of these vaccines have been tested for immunogenicity and protective efficacy in mouse and guinea pig models of low-dose pulmonary tuberculosis. In addition, alternative routes of tuberculosis vaccine delivery (e.g. oral, respiratory, gene gun) and various combinations of priming or boosting an experimental vaccine with bacille Calmette Guerin have been examined in relevant animal models. One of the most promising of these vaccines is currently in Phase I trials in human subjects, and others are expected to follow in the near future. This review will summarise the most recent progress made toward the development and preclinical evaluation of novel vaccines for human tuberculosis. PMID- 12782055 TI - Development of vaccines to control bovine tuberculosis in cattle and relationship to vaccine development for other intracellular pathogens. AB - Vaccination of cattle against bovine tuberculosis could be an important strategy for the control of disease either where there is a wildlife reservoir of Mycobacterium bovis infection or in developing countries where it is not economically feasible to implement a 'test and slaughter' control program. Advances in the understanding of protective immune responses to M. bovis infection in cattle and the advent of new molecular biological techniques, coupled with the sequencing of the M. bovis genome have provided opportunities for the rational development of improved tuberculosis vaccines. A number of new tuberculosis vaccines including attenuated M. bovis strains, killed mycobacteria, protein and DNA vaccines are under development and many are being assessed in cattle. Recent results have revealed several promising vaccine candidates and vaccination strategies. Ways of distinguishing between vaccinated and infected cattle are becoming available and the possibility of new approaches to the eradication of tuberculosis from domestic livestock is discussed. Similarities between the mechanisms of protective immunity against M. bovis and against other intracellular parasites continue to be found and discoveries from vaccine studies on bovine tuberculosis may provide helpful insights into requirements for vaccines against other intracellular pathogens. PMID- 12782056 TI - Traditional and novel approaches to flavivirus vaccines. AB - Yellow fever, dengue, Japanese encephalitis and tick-borne encephalitis viruses are the medically most important members of the Flavivirus genus composed primarily of arboviruses. In this paper, we review the commercially available traditional flavivirus vaccines against yellow fever, Japanese encephalitis, and tick-borne encephalitis, as well as modern approaches to flavivirus vaccines. Formalin inactivation technology has been employed to produce killed vaccines. Flaviviruses have been attenuated by multiple passages in animal tissues and cell cultures to produce empirical live attenuated vaccines. The use of traditional methods is being pursued to develop vaccines against other flavivirus diseases, such as dengue, and to improve existing vaccines, such as for Japanese encephalitis. With the recent development of infectious clones, rational approaches to attenuated flavivirus vaccines have employed the introduction of specific mutations into wild type viruses and chimerisation between different viruses. Novel methods for delivery of live vaccines, such as inoculation of infectious DNA or RNA, have been described. Other approaches, such as the construction of protein subunit, expression vector-based and naked DNA vaccines, have been proposed to create alternate vaccine candidates. PMID- 12782057 TI - Vaccine research efforts for filoviruses. AB - Ebola and Marburg viruses belong to the family Filoviridae, and cause acute, frequently fatal, haemorrhagic fever in humans and non-human primates. No vaccines are available for human use. This review describes the status of research efforts to develop vaccines for these viruses and to identify the immune mechanisms of protection. The vaccine approaches discussed include DNA-based vaccines, and subunit vaccines vectored by adenovirus, alphavirus replicons, and vaccinia virus. PMID- 12782058 TI - Strategies in cancer vaccines development. AB - The recent definition of tumour-specific immunity in cancer patients and the identification of tumour-associated antigens have generated renewed enthusiasm for the application of immune-based therapies for the treatment of malignancies. Recent developments in cancer vaccines have also been based on an improved understanding of the cellular interactions required to induce a specific anti tumour immune response. Consequently, a number of cancer vaccines have entered clinical trials. Targeting broad-spectrum tumour-associated antigens has emerged as a strategy to lower the risk of tumour escape due to the loss of specific nominal antigen. Amongst the most challenging of tumour-associated antigens to which to target in active specific immunotherapy applications are carbohydrate antigens. As carbohydrates are intrinsically T-cell-independent antigens, more novel approaches are perhaps needed to drive specific-T-cell-dependent immune responses to carbohydrate antigens. In this context peptide mimetics of core structures of tumour-associated carbohydrate antigens might be developed to augment immune responses to these broad-spectrum antigens. PMID- 12782059 TI - Rational antibacterial vaccine design through genomic technologies. AB - After 200 years of practice, vaccinology has proved to be very effective in preventing infectious diseases. However, several human and animal pathogens exist for which vaccines have not yet been discovered. As for other fields of medical sciences, it is expected that vaccinology will greatly benefit from the emerging genomics technologies such as bioinformatics, proteomics and DNA microarrays. In this review, the potential of these technologies will be illustrated taking into account part of the research activities currently in progress in our laboratories. In particular, I will describe the identification of new vaccine candidates against Meningococcus B through high-throughput cloning and expression of meningococcal antigens selected by: (i) in silico analysis of genome sequence; and (ii) transcriptome analysis of bacteria adherent to epithelial cells. In addition, I will show how the combination of high-throughput cloning and expression technology with two-dimensional gel/mass spectrometry led us to the elucidation of Chlamydia pneumoniae surface protein subproteome and to the identification of potential vaccine candidates. PMID- 12782060 TI - Helminth vaccines: from mining genomic information for vaccine targets to systems used for protein expression. AB - The control of helminth diseases of people and livestock continues to rely on the widespread use of anti-helminthic drugs. However, concerns with the appearance of drug resistant parasites and the presence of pesticide residues in food and the environment, has given further incentive to the goal of discovering molecular vaccines against these pathogens. The exponential rate at which gene and protein sequence information is accruing for many helminth parasites requires new methods for the assimilation and analysis of the data and for the identification of molecules capable of inducing immunological protection. Some promising vaccine candidates have been discovered, in particular cathepsin L proteases from Fasciola hepatica, aminopeptidases from Haemonchus contortus, and aspartic proteases from schistosomes and hookworms, all of which are secreted into the host tissues or into the parasite intestine where they play important roles in host-parasite interactions. Since secreted proteins, in general, are exposed to the immune system of the host they represent obvious candidates at which vaccines could be targeted. Therefore, in this article, we consider the potential values and uses of algorithms for characterising cDNAs amongst the collated helminth genomic information that encode secreted proteins, and methods for their selective isolation and cloning. We also review the variety of prokaryotic and eukaryotic cell expression systems that have been employed for the production and downstream purification of recombinant proteins in functionally active form, and provide an overview of the parameters that must be considered if these recombinant proteins are to be commercialised as vaccine therapeutics in humans and/or animals. PMID- 12782061 TI - T cell epitope identification for bovine vaccines: an epitope mapping method for BoLA A-11. AB - T cell responses play an important role in immunity to parasites and other microbial agents of infectious diseases, therefore a number of T cell-directed vaccines are in development. Computer-driven algorithms that facilitate the discovery of T cell epitopes from protein and genome sequences are now being used to accelerate preclinical studies of human vaccines. Similar tools are not yet available for predicting T cell epitopes for animal vaccines, but there may be sufficient data available to begin the process of compiling the algorithms. We describe the construction of a novel mathematical 'matrix' that describes the properties of bovine major histocompatibility complex (BoLA) system antigen (BoLA) A-11 peptide ligands, developed for use with EpiMatrix, an existing T cell epitope-mapping algorithm. An alternative means of developing BoLA matrices, using the pocket profile method, is also discussed. Matrices such as the one described here may be used to develop T cell epitope-mapping tools for cattle and other ruminants. Epitope-mapping algorithms offer a significant advantage over other methods of epitope selection, such as the screening of synthetic overlapping peptides, because high throughput screening can be performed in silico, followed by ex vivo confirmatory studies. Furthermore, using epitope mapping algorithms, putative T cell epitopes can be derived directly from genomic sequences, allowing researchers to circumvent labor-intensive cloning steps in the genome-to-vaccine discovery pathway. PMID- 12782062 TI - The effectiveness and limitations of immune memory: understanding protective immune responses. AB - Immune memory is the foundation of the practise of vaccination. Research on the molecular and cellular events leading to generation and development of memory T and B lymphocytes explain why there are heightened secondary immune responses after an initial encounter with antigen. In this review, we discuss how clonal expansion, targeted tissue localisation, more efficient antigen recognition and more proficient effector functions contribute to the improved effectiveness of memory cells. Despite the enhanced efficacy of memory cells and the recall immune response, there are numerous experimental and empirical examples in which protection provided by vaccines are short-lived, particularly against pathogens that replicate and cause pathology at their site of entry. In the absence of active immune effector activities, the ability of memory cells to respond quickly enough to control this type of infection is limited. The protective efficacy of bovine herpes virus-1 vaccines in experimental and field challenge conditions are used to illustrate the concept that full protection from disease conferred by vaccination requires the presence of active immune effector mechanisms. Thus, regardless of the many successful technological advances in vaccine design and better understanding of mechanisms underlining induction of memory responses by vaccination, we should recognise that vaccine immunoprophylaxis has limitations. Expectations for vaccines should be realistic and linked to the understanding of host immune responses and knowledge regarding the pathogen and disease pathogenesis. PMID- 12782063 TI - Configural information is processed differently in perception and recognition of faces. AB - Several previous studies have stressed the importance of processing configural information in face recognition. In this study the perception of configural information was investigated. Large overestimations were found when the eye-mouth distance and the inter-eye distance had to be estimated. Whereas configural processing is disrupted when inverted faces have to be recognized the perceptual overestimations persisted when faces were inverted. These results suggest that processing configural information is different in perceptual as opposed to recognition tasks. PMID- 12782064 TI - Spatial-frequency spectra of printed characters and human visual perception. AB - It is well known that certain spatial frequency (SF) bands are more important than others for character recognition. Solomon and Pelli [Nature 369 (1994) 395 397] have concluded that human pattern recognition mechanism is able to use only a narrow band from available SF spectrum of letters. However, the SF spectra of letters themselves have not been studied carefully. Here I report the results of an analysis of SF spectra of printed characters and discuss their relationship to the observed band-pass nature of letter recognition. PMID- 12782065 TI - Ontogenic changes of kynurenine aminotransferase I activity and its expression in the chicken retina. AB - Kynurenine aminotransferases are key enzymes for the synthesis of kynurenic acid (KYNA), an endogenous glutamate receptor antagonist. The study described here examined ontogenic changes of kynurenine aminotransferase I (KAT I) activity and its expression in the chicken retina. KAT I activity measured on embryonic day 16 (E16) was significantly higher than at all other stages (E12, P0 and P7). Double labeling with antibodies against glutamine synthetase showed that on P7 KAT I was expressed in Muller cell endfeet and their processes in the inner retina. Since KAT I activity is high in the late embryonic stages, it is conceivable that it plays a neuromodulatory role in the retina during the late phase of embryogenesis. PMID- 12782066 TI - Further evidence that chick eyes use the sign of blur in spectacle lens compensation. AB - Young animals compensate for defocus imposed by positive or negative spectacle lenses by adjusting the elongation rate of their vitreous chambers, thus matching the length of the eye with the focal length of the eye's optics combined with the spectacle lenses. The ability to compensate for either negative or positive lenses could rely on the ability to distinguish between myopic and hyperopic blur, or it could rely on the fact that positive lenses would bring nearby objects into focus, thereby reducing the amount of blur, whereas negative lenses would not. This study asks whether eyes emmetropize using the magnitude of blur or the sign of blur as a directional cue. We fitted chick eyes with positive lenses while imposing a substantial amount of blur, either (a) by having them wear lenses only when restrained in the center of a cylinder, the walls of which were beyond their far-point or (b) by having them wear mild diffusers over positive lenses. We found good refractive compensation in both situations in a large number of birds. Furthermore, we found that mild diffusers worn on top of positive lenses differentially affected the two ocular components of refractive compensation: there was less choroidal thickening, but more inhibition of ocular elongation, compared to wearing positive lenses alone. These findings argue both that the eye can discern the sign of the blur and that choroidal and ocular elongation components of the refractive compensation do not respond identically to visual inputs. PMID- 12782067 TI - Subjective contours and binocular rivalry suppression. AB - Binocular rivalry probably involves distributed neural processes, some responsible for dominance, others for suppression and still others for fluctuations in perception. Focusing on the suppression process, the present study asks whether neural events underlying rivalry suppression take place prior to, or subsequent to those underlying the synthesis of subjective contours. Specifically, we examined whether (i) a subjective contour could prematurely return a suppressed target to dominance and (ii) whether suppression of a Kanizsa type inducer precludes the formation of a subjective contour. Suppression durations were not abbreviated by the subjective contour, but suppression did prevent the formation of a subjective contour. Evidently suppression precedes the synthesis of subjective contours in the visual processing hierarchy. PMID- 12782068 TI - Distractor heterogeneity, attention, and color in visual search. AB - Experiments were designed to investigate the effects of set size and variation in the chromaticity of distractor stimuli on thresholds for detecting a target stimulus that differed from distractors only in chromaticity. Distractor chromaticities were selected from a line in the isoluminant color plane and targets were selected from lines approximately orthogonal to the distractor line. With uniform distractors thresholds increased with set size as predicted by a signal detection model. When targets and distractors were selected from lines parallel to the Cardinal directions in color space, thresholds were lower with variable distractors than with uniform distractors and variations in the location of the target along the distractor line had no effect on threshold. Results with diagonally oriented distractor lines were similar. Results suggest that many pairs of orthogonal directions in the isoluminant color plane represent independent color coding mechanisms that mediate search. Results also show that information in independent color coding mechanisms tuned to orthogonal directions in the isoluminant plane can be combined to facilitate detection of the target. PMID- 12782069 TI - Eye movements provide the extra-retinal signal required for the perception of depth from motion parallax. AB - It has been unclear whether the perception of depth from motion parallax is an entirely visual process or whether it requires extra-retinal information such as head movements, vestibular activation, or eye movements. Using a motion aftereffect and static test stimulus technique to eliminate visual cues to depth, this psychophysical study demonstrates that the visual system employs a slow eye movement signal, optokinetic response (OKR) in particular, for the unambiguous perception of depth from motion parallax. A vestibular signal, or vestibularly driven eye movement signal is insufficient for unambiguous depth from motion parallax. Removal of the OKR eye movement signal gives rise to ambiguous perceived depth in motion parallax conditions. Neurophysiological studies suggest a possible neural mechanism in medial temporal and medial superior temporal cortical neurons that are selective to depth, motion, and direction of eye movement. PMID- 12782070 TI - Early binocular vision in human infants: limitations on the generality of the Superposition Hypothesis. AB - The Superposition Hypothesis states that the binocular vision of newborn infants blends together the monocular visual responses of the two eyes, even when the visual stimulus evokes binocular rivalry in adults. According to the Superposition Hypothesis, this blending is replaced by binocular rivalry after the emergence of stereopsis [Neonate Cognition: Beyond the Blooming, Buzzing Confusion, Erlbaum, Hillsdale, NJ, 1985, p. 37; Early Visual Development Normal and Abnormal, Oxford University Press, New York, 1993, p. 201]. The main evidence for the Superposition Hypothesis is a preferential looking experiment [Vision Res., 26 (3) (1986) 501], in which 8-12-week-old infants fixated a rivalrous (for adults) dichoptic plaid, in preference over a fusible grating of parallel lines. This report describes our attempt to repeat that important experiment. Infant stereopsis emerged at 8.6 weeks under our conditions, but infants did not preferentially fixate the dichoptic plaid at any age between age 5 and 16 weeks. Control experiments showed that our result was not due to technical differences between their experiment and ours (red/green vs. polarizing glasses, the use of a fixation point, or the infant observation apparatus). Therefore, blending of the visual responses to rivalrous (for adults) stimuli is not a general feature of the pre-stereoptic infant visual system. PMID- 12782071 TI - How many positions can we perceptually encode, one or many? AB - Here we show that our sensitivity for discriminating relative position across the visual field is limited. In experiment 1 we show that we are much worse at detecting a texture defined by the relative position of elements within an array than would be expected if we had access to multiple estimates of relative position across the visual field. In experiment 2 we show that human performance is impaired for positional judgments when there is uncertainty as to which of a number of possible elements is misaligned. This impairment is greater than one would expect from an ideal observer model and greater than that found for a comparable task involving orientation. It is consistent with positional thresholds being determined by only one estimate of relative position. In experiment 3 we estimate the number of suprathreshold positional signals that can be pooled at the same time across the visual field using a standard summation variance paradigm. The results suggest that the human visual system is limited to one estimate of position, but additional estimates can be built up serially over time; however, this process is slow and probably cognitive in nature. These experiments taken as a whole suggest that only one estimate of relative position (i.e. relative to a predefined reference) at a time is accessible at the perceptual level. PMID- 12782072 TI - Visual motion detection in patients with absent vestibular function. AB - Labyrinthine defective subjects (LDS) experience oscillopsia during head movements due to the absence of the vestibulo-ocular reflex (VOR). The purpose of this study was to compare horizontal and vertical visual motion detection in LDS during (i) body-stationary and (ii) horizontal whole-body oscillation conditions. Twelve LDS and controls detected the onset of drift direction of a grating that moved with accelerating velocity. Thresholds were raised in the patient group in both conditions. The loss of the VOR per se cannot explain raised thresholds in the body-stationary condition nor during whole-body (horizontal) oscillation with vertical grating motion. Findings indicate changes in visual processing that make LDS less sensitive to visual motion. It is postulated that these changes are due to adaptive mechanisms involved to reduce oscillopsia. PMID- 12782073 TI - Purification and primary structure determination of Tf4, the first bioactive peptide isolated from the venom of the Brazilian scorpion Tityus fasciolatus. AB - In the present study Tityus fasciolatus crude venom toxicity was evaluated and we also report the purification and characterization of a 6.6 kDa neurotoxin isolated from T. fasciolatus venom. This new toxin, named Tf4, has a molecular mass of 6614Da and its primary structure is homologous to TbIT-I from T. bahiensis and TsTX-VI and TsNTxP from T. serrulatus. Tf4 delays frog sodium channel inactivation reversibly, but it is non-toxic to mammals or crustaceans. An attempt to identify the residues responsible for the partial loss of toxicity in Tf4 was carried out based on homology modeling and sequence comparison. PMID- 12782074 TI - Paralytic toxicity in the ribbon worm Cephalothrix species (Nemertea) in Hiroshima Bay, Hiroshima Prefecture, Japan and the isolation of tetrodotoxin as a main component of its toxins. AB - Paralytic toxicity of ribbon worms ("himomushi" in Japanese), identified as undescribed species of the genus Cephalothrix, found on the surface of the shells of cultured oysters in Hiroshima Bay, Hiroshima Prefecture was examined between April 1998 and December 2001. The toxicity study showed that all of specimens were found to contain toxins with strong paralytic action in mice; the highest toxicity (as tetrodotoxin, TTX) was 25,590 mouse units (MU) per gram for whole body throughout the monitoring period. The main toxic component of this himomushi toxin (HMT) was isolated from a pooled specimen (390 g; total toxicity 2,897,000MU) by a method that consisted of treatment with activated charcoal, chromatography on Bio-Gel P-2 and Bio-Rex 70 (H+ form), and finally crystallization from an acidified methanolic solution. The recrystallized toxin showed a specific toxicity of 3520MU/mg. This toxin showed (M+H)+ and (M+H H(2)O)+ ion peaks at m/z 320 and 302, respectively, by electrospray ionization mass spectrometry (ESI-MS). The absorption band at 3353, 3235, 1666, 1612 and 1076 cm(-1) were observed in infrared spectrum of this toxin. This spectrum was indistinguishable from that of TTX. The 1H-NMR spectrum for the recrystallized toxin was the same as that for TTX. The pair of doublets centered at 2.33 (J=10.0Hz) and 5.48 ppm (J=10.0Hz) which are characteristic of TTX, were shown to be coupled by double irradiation. Furthermore, by gas chromatography-mass spectrometry (GC-MS) of the alkali-hydrolyzate of this toxin indicated the presence of quinazoline skeleton (C9-base) specific to TTX. PMID- 12782075 TI - Molecular cloning and characterization of Phoneutria nigriventer toxins active on calcium channels. AB - The aim of the present study was the molecular cloning of toxins active on calcium channels expressed by the spider Phoneutria nigriventer. Clones encoding the toxins Pn3-3A, Pn3-4A, Tx3-5, Pn3-5A, Tx3-6, Pn3-6A and Pn3-6B were identified from a cDNA library derived from the venom gland of this spider, revealing toxins of 49, 76, 45, 39, 55 and 58 amino acids residues, respectively, with polypeptide precursors being composed of three major portions: a signal peptide, a propeptide and finally, the mature toxin. A high degree of homology with the amino acid sequence was found between Pn3-3A and the neurotoxin Tx3-3 (identity of 79%), and between Pn3-4A and the neurotoxin Tx3-4 (identity of 95%). The deduced amino acid sequence for the mature polypeptides Tx3-5 and Tx3-6 confirms the polypeptide sequence previously published for these neurotoxins. In addition, the toxin Pn3-5A showed 58% identity to the Tx3-5 amino acid sequence, and the toxins Pn3-6A and Pn3-6B showed 85 and 33% identity, respectively, to the Tx3-6 amino acid sequence. PMID- 12782076 TI - Cloning and characterization of a blood coagulation factor IX-binding protein from the venom of Trimeresurus stejnegeri. AB - A blood coagulation factor IX-binding protein (TSV-FIX-BP) was isolated from the snake venom of Trimeresurus stejnegeri. On SDS-polyacrylamide gel electrophoresis, TSV-FIX-BP showed a single band with an apparent molecular weight of 23,000 under non-reducing conditions, and two distinct bands with apparent molecular weights of 14,800 and 14,000 under reducing conditions. cDNA clones containing the coding sequences of TSV-FIX-BP were isolated and sequenced to determine the structure of the precursors of TSV-FIX-BP subunits. The deduced amino acid sequences of two subunits of TSV-FIX-BP were confirmed by N-terminal protein sequencing and trypsin-digested peptide mass fingerprinting. TSV-FIX-BP was a non-enzymatic C-type lectin-like anti-coagulant. The anti-coagulant activity of TSV-FIX-BP was mainly caused by its dose dependent interaction with blood coagulation factor IX but not with blood coagulation factor X. PMID- 12782077 TI - Inhibition of nuclear protein phosphatase activity in mouse hepatocytes by the cyanobacterial toxin microcystin-LR. AB - Microcystin-LR (MCLR) is a cyanobacterial hepatotoxin and protein phosphatase inhibitor that contaminates water reservoirs worldwide. MCLR localizes to the cytosol of hepatocytes, however, immunohistochemical studies indicate that it accumulates in the nucleus. MCLR toxicosis is associated with decreased hepatic protein phosphatase activity, but effects in nuclear protein phosphatase activity have not been investigated. Balb/c mice were given lethal (100 microg/kg) or sublethal (12, 23 and 45 microg/kg) i.p. doses of MCLR and hepatic nuclear extracts were analyzed for protein phosphatase 1 and 2A activity. There was profound inhibition of nuclear protein phosphatase activity within 50 min of lethal dosing, however an inhibition was not detected with sublethal doses. MCLR immunohistochemistry revealed widespread lobular staining in the lethal group and centrilobular staining in the sublethal groups. At the cellular level there was nuclear and cytoplasmic staining of equal intensity. As an indicator of nuclear protein phosphatase activity, the phosphorylation of p53, a nuclear phosphoprotein and known substrate for protein phosphatases 1 and 2A, was evaluated. Balb/c mice were treated with sublethal doses of MCLR or saline vehicle after induction of hepatic p53 by the DNA damaging agent diethylnitrosamine (DEN). P53 was immunoprecipitated and probed with phosphoserine specific antibodies by Western blotting. There was greater phosphoserine reactivity of p53 protein in animals treated with MCLR relative to saline treated controls, consistent with increased phosphorylation of serine sites. It is concluded that an interaction of this toxin with nuclear protein phosphatases occurs within 50 min of lethal dosing, which leads to a profound inhibition of enzymatic activity. Even sublethal doses of MCLR that do not result in significant inhibition of activity in bulk nuclei, result in detectable changes in phosphorylation of p53. PMID- 12782078 TI - Oral and intraperitoneal acute toxicity studies of yessotoxin and homoyessotoxins in mice. AB - The acute toxicity of yessotoxin (YTX), homoyessotoxin (homoYTX) and 45-hydroxy homoyessotoxin (45-OH-homoYTX) has been studied in comparison to that of okadaic acid (OA), the main diarrhogenic toxin, both after intraperitoneal (i.p.) and oral administration. After i.p. administration, homoYTX and YTX showed similar lethality (LD(50)=444 microg/kg and 512 microg/kg), higher than that of OA (LD(50)=225 microg/kg), while 750 microg/kg of 45-OH-homoYTX did not cause death. OA induced the already known toxic signs: before death, mice were motionless and cyanotic; small intestine and liver damage were shown at post-mortem. Mice treated with YTX and homoYTX were restless and jumped before death; necroscopy did not show major changes. After oral treatment, 2 mg/kg of OA induced diarrhoea and body weight loss, causing 4/5 deaths; necroscopy and/or histology revealed degenerative lesions to small intestine, forestomach and liver (confirmed by increased plasma transaminase), but no myocardium alterations. On the contrary, the oral treatment with YTX (1 and 2 mg/kg) and its derivatives (1 mg/kg) did not cause any death or signs of toxicity, except some ultrastructural myocardiocyte alterations, adjacent to capillaries, such as cytoplasmic protrusions (YTX, 1 and 2 mg/kg), fibrillar alteration (YTX, 1 mg/kg) or mitochondria assemblage (45-OH homoYTX). Altogether, our data show that YTX and its derivatives are less toxic than OA after acute oral and i.p. treatments, at doses which may represent up to 100 times of the possible human daily intake. PMID- 12782079 TI - Cloning and characterization of a novel neurotoxin from the sea anemone Anthopleura sp. AB - A full-length cDNA of neurotoxin (Hk2a) was isolated by RT-PCR of total RNA isolated from tentacles of Anthopleura sp. using degenerate oligonucleotide primers and 3',5'-RACE. The cDNA sequence of Hk2a encoded a polypeptide of 47 amino acids, which lacks a typical N-terminal signal sequences commonly found in proteins that are secreted via endoplasmic reticulum-Golgi pathway, indicating the possibility of secretion via a non-classical pathway. The neurotoxin has a predicted molecular mass of 4.8 kDa and a pI value of 7.62. The amino acid sequence of Hk2a is very similar to Anthopleurin C (Ap-C) and Neurotoxin I (Af I), and shares 95% amino acid sequence similarity to Ap-C. The coding region for the matured Hk2a toxin was cloned into the thioredoxin (TRX) fusion expression vector (pTRX) for the fusion expression in Escherichia coli. The recombinant polypeptide of Hk2a (rHk2a) was purified by the affinity chromatography, 15 mg/l of rHk2a was obtained after the digestion with protease 3C and further purification. The molecular weight of rHk2a (5.078 kDa) obtained by MALDI-TOF was very close to that (5Da) calculated from the sequence. The results of the UV circular dichroism spectra of rHk2a indicates that its secondary structure is similar to that of Ap-B (), having 61.7% beta-sheet and no alpha-helix. Investigation on pharmacological effects of rHk2a in vitro was undertaken, and it was found that LD(50) of rHk2a was 1.4 mg/kg on NIH mice (i.p.). The rHk2a was demonstrated to increase contracting activity on isolated SD rat atria with the enhancing degree reaching 343.5+/-160.5%. The increase in contractile amplitude reached a plateau value within 3-5 min after addition of this toxin. PMID- 12782080 TI - Anti-mitotic activity towards sea urchin embryos in extracts from the marine haptophycean Phaeocystis pouchetii (Hariot) Lagerheim collected along the coast of northern Norway. AB - The marine bloom-forming alga Phaeocystis pouchetii is suspected to produce some toxic compound responsible for reduced growth, fecundity and survival of other marine organisms. Sea urchin early development was used as a model to investigate the degree and nature of toxicity. Colonial cells of P. pouchetii were collected during its spring-bloom along the coast of northern Norway and maintained in culture for a short period of time in order to evaluate the concentration of toxic compounds present inside the cells or excreted to the surrounding seawater medium. Cells were harvested by filtration and toxins were extracted separately from the collected cells and the filtrate using organic solvents. We found that extracts from the filtered seawater at a concentration corresponding to 9.0 x 10(5) cells ml(-1) completely blocked cell divisions in embryos of the sea urchin Sphaerechinus granularis, whereas extracts from intact algal cells were only mildly cytotoxic. When the extracts from seawater culture medium were purified by RP-HPLC, cytotoxic activity towards S. granularis embryos was recovered in three consecutive fractions. Moreover, unfertilised eggs incubated in the active HPLC fractions became unproductive, whereas incubation of sperm gave a reduced fertilisation rate. This anti-proliferative effect was further characterized by immunofluorescence staining of sea urchin embryos. DNA labelling revealed that incubating sea urchin embryos in the purified algal extracts inhibited both pronuclei migration and fusion. Incorporation and detection of the DNA-base analogue 5-bromo-2-deoxyuridine showed that DNA-replication was blocked. Furthermore, staining of alpha-tubulin subunits demonstrated that embryonic tubulin organisation was altered. We conclude that P. pouchetii produce some anti mitotic compound, and that senescent colonial cells to a great extent excrete this compound to their surroundings. PMID- 12782081 TI - Pulmonary gene expression profiling of inhaled ricin. AB - Aerosol exposure to ricin causes irreversible pathological changes of the respiratory tract resulting in epithelial necrosis, pulmonary edema and ultimately death. The pulmonary genomic profile of BALB/c mice inhalationally exposed to a lethal dose of ricin was examined using cDNA arrays. The expression profile of 1178 mRNA species was determined for ricin-exposed lung tissue, in which 34 genes had statistically significant changes in gene expression. Transcripts identified by the assay included those that facilitate tissue healing (early growth response gene (egr)-1), regulate inflammation (interleukin (IL)-6, tristetraproline (ttp)), cell growth (c-myc, cytokine-inducible SH2-containing protein (cish)- 3), apoptosis (T-cell death associated protein (tdag)51, pim-1) and DNA repair (ephrin type A receptor 2 (ephA2)). Manipulation of these gene products may provide a means of limiting the severe lung damage occurring at the cellular level. Transcriptional activation of egr-1, cish-3, c-myc and thrombospondin (tsp)-1 was already apparent when pathological and physiological changes were observed in the lungs at 12 h postexposure. These genes may well serve as markers for ricin-induced pulmonary toxicity. Ongoing studies are evaluating this aspect of the array data and the potential of several genes for clinical intervention. PMID- 12782082 TI - Inflammatory oedema induced by phospholipases A2 isolated from Crotalus durissus sp. in the rat dorsal skin: a role for mast cells and sensory C-fibers. AB - The ability of the phospholipases A(2) (PLA(2)s) from Crotalus durissus cascavella, Crotalus durissus collilineatus and Crotalus durissus terrificus venoms and crotapotin to increase the vascular permeability in the rat skin as well as the contribution of both mast cells and sensory C-fibers have been investigated in this study. Vascular permeability was measured as the plasma extravascular accumulation at skin sites of intravenously injected 125I-human serum albumin. Intradermal injection of crotalic PLA(2)s (0.05-0.5 microg/site) in the rat skin resulted in dose-dependent increase in plasma extravascular whereas crotapotin (1 microg/site) failed to affect this response. Co-injection of crotapotin (1 microg/site) did not modify the increased vascular permeability induced by the PLA(2)s (0.05-0.5 microg/site). Previous treatment (30 min) of the animals with cyproheptadine (2 mg/kg, i.p.) markedly reduced PLA(2) (0.5 microg/site)-induced oedema. In rats treated neonatally with capsaicin to deplete neuropeptides, the plasma extravasation induced by all PLA(2)s (0.5 microg/site) was also significantly reduced. Similarly, the tachykinin NK(1) receptor antagonist SR140333 (1nmol/site) significantly reduced the PLA(2)-induced oedema. In addition, the combination of SR140333 with cyproheptadine further reduced the increased plasma extravasation by PLA(2) from C. d. cascavella venom, but not by PLA(2) from C. d. terrificus and C. d. collilineatus venoms. Our results suggest that increase in skin vascular permeability by crotalic PLA(2)s is mediated by activation of sensory C-fibers culminating in the release of substance P, as well as by activation of mast cells which in turn release amines such as histamine and serotonin. PMID- 12782083 TI - In vitro hemolytic activity of Lonomia obliqua caterpillar bristle extract on human and Wistar rat erythrocytes. AB - Human accidental envenomation caused by skin contact with the bristles of Lonomia obliqua caterpillar causes coagulation and fibrinolysis disorders. Alterations of hematologic parameters are observed only in severe cases of envenomation, but with no clinical evidence of intravascular hemolysis. However, since we have observed intravascular hemolysis in preliminary studies using Wistar rats as an experimental model for investigating L. obliqua envenomation, the objective of the present study was to investigate the in vitro hemolytic activity of the bristle extract of L. obliqua caterpillars on human and rat erythrocytes. Our results showed that the bristle extract has indirect and direct hemolytic activity on human and rat erythrocytes, although direct hemolytic activity was only observed at higher bristle extract concentrations. We also observed that the bristle extract has a proteolytic activity on band 3 of human and rat erythrocyte membranes. Thus, crude L. obliqua bristle extract was found to contain at least two components with hemolytic activity on erythrocytes, a phospholipase enzyme and another protein with a direct activity on the erythrocyte membrane. PMID- 12782084 TI - Production of Abrus pulchellus ribosome-inactivating protein from seeds callus culture. AB - Ribosome inactivating proteins (RIPs) were isolated from callus culture that were established from seed explants of Abrus pulchellus. Cotyledon segments of immature seeds were inoculated in basal medium MS supplemented with different concentrations of auxin (2,4-D), citokinin (kinetin and BA) and sucrose in order to determine the best callus induction. A. pulchellus type 2 RIP (pulchellin) expression was monitored in callus cultures by RT-PCR and biological activity. The calli obtained after 35 days were freeze dried, macerated and submitted to extraction of total RNA and proteins (0.1 M Tris-HCl pH 7.6 buffer, containing 0.15 M NaCl, 3 h at room temperature). A specific DNA fragment codifying the A chain pulchellin was amplified from callus RNA suggesting the presence of the protein. This was confirmed in the calli crude extract that showed haemagglutinating activity against rabbit blood cells and a high intraperitoneal toxicity to mice. The crude extract was also submitted to affinity chromatography on a Sepharose-4B column. The retained protein, peak released by 0.1 M galactose, appeared to be composed of two main bands in polyacrylamide gel electrophoresis, in denaturating conditions, with a similar pattern to that obtained with seeds. PMID- 12782085 TI - Preclinical assessment of the ability of polyvalent (Crotalinae) and anticoral (Elapidae) antivenoms produced in Costa Rica to neutralize the venoms of North American snakes. AB - Polyvalent (Crotalinae) and anticoral (Elapidae) antivenoms produced by Instituto Clodomiro Picado, Costa Rica, were assessed for their ability to neutralize various toxic activities of the venoms of North American snakes of the genera Crotalus, Agkistrodon and Micrurus, in assays involving preincubation of venom and antivenom. When the intraperitoneal route of injection was utilized, polyvalent (Crotalinae) antivenom was effective in the neutralization of the venoms of Crotalus atrox, Crotalus adamanteus, Crotalus viridis viridis, Crotalus horridus atricaudatus, Agkistrodon contortrix contortrix and Agkistrodon piscivorus piscivorus, whereas the venom of Crotalus scutulatus was not neutralized. When the intravenous route was used, results differed depending on the "challenge dose" of venom employed. Polyvalent antivenom neutralized all venoms when mice were challenged with 2 LD(50)s of venom. When 5 LD(50)s were used, antivenom neutralized the venoms of C. atrox, C. adamanteus, C. v. viridis and C. h. atricaudatus, being ineffective in the neutralization of C. scutulatus, A. c. contortrix and A. p. piscivorus. Polyvalent antivenom was effective in the neutralization of hemorrhagic and myotoxic activities of all venoms studied. It also neutralized coagulant activity of C. adamanteus venom, whereas most of the venoms were devoid of clotting activity on plasma in vitro. Moreover, it neutralized defibrinating activity of the only three venoms that induced this effect (i.e. C. adamanteus, A. c. contortrix and A. p. piscivorus). Anticoral (Elapidae) antivenom neutralized lethality induced by the venom of Micrurus fulvius, using either the intravenous or the intraperitoneal routes of injection. Moreover, it neutralized myotoxic effect of this venom as well. It is concluded that polyvalent antivenom neutralizes lethality and other activities of most of the crotaline venoms tested. However, since it is ineffective in neutralizing the lethal effect of C. scutulatus venom, it is suggested that a venom containing presynaptically-active neurotoxic phospholipases A(2) related to "mojave toxin" needs to be introduced in the immunizing mixture in order to increase the neutralizing scope of this product in North America. Anticoral antivenom is highly effective in the neutralization of the venom of M. fulvius. PMID- 12782086 TI - Inhibition of mammary carcinoma cell proliferation in vitro and tumor growth in vivo by bee venom. AB - The possible tumor growth- and metastasis-inhibiting effects of bee venom in mice and in tumor cell cultures were studied. The tumor was a transplantable mammary carcinoma (MCa) of CBA mouse. Intravenous administration of bee venom to mice significantly reduced the number of metastases in the lung. However, subcutaneous administration of bee venom did not reduce the number of lung metastases, indicating that the antitumor effect of the venom could be highly dependent on the route of injection as well as close contact between the components of the venom and the tumor cells, as was shown by in vitro studies on MCa cells. We also observed variations in immunological parameter induced by bee venom. We proposed that bee venom has an indirect mechanism of tumor growth inhibition and promotion of tumor rejection that is based on stimulation of the local cellular immune responses in lymph nodes. Apoptosis, necrosis, and lysis of tumor cells are other possible mechanisms by which bee venom inhibits tumor growth. PMID- 12782087 TI - The use of Lepidium sativum in a plant bioassay system for the detection of microcystin-LR. AB - Toxin-producing cyanobacteria pose a worldwide health threat to humans and animals due to their increasing presence in both drinking and recreational waters. Detection of microcystins in water generally relies on specialised equipment and a delay of several days for transport and analysis. Little work has, however, been done on establishing a simple, cost-effective and sensitive plant bioassay for the detection of microcystin-LR (MCLR) in water at the WHO Tolerable Daily Intake guideline level of 1 microg/l. We investigated the effect of a MCLR extract at 1 and 10 microg/l on the growth of Lepidium sativum over 6 days. Exposure to 10 microg/l MCLR resulted in a significant decrease in root and leaf lengths and fresh weights of seedlings when compared to the controls. These results were consistent with seedlings exposed to pure MCLR at 10 microg/l. Seedlings exposed to 1 microg/l MCLR showed a significant decrease in root development from day 2 to day 6. Glutathione S-transferase and glutathione peroxidase activities were also significantly raised in plants from days 5 and 4, respectively, at both toxin levels investigated. PMID- 12782088 TI - Australian scorpion stings: a prospective study of definite stings. AB - There is little information on scorpion stings in Australia. The aim of this study is to describe the circumstances and clinical effects of stings by Australian scorpions. Cases of scorpion stings were collected prospectively from calls and presentations to Australian poison information centres and emergency departments from February 2000 to April 2002. Only definite scorpion stings where the scorpion was immediately collected and expertly identified were included. There were 95 patients, 33 males and 62 females, with a mean age of 32 (SD 19.5; range 1-71) and 23 children (age<15 years). Three families of scorpions caused all stings: Buthidae (79), Bothruiridae (11, all Cercophonius spp.) and Urodacidae (five, all Urodacus spp.). The majority of stings (76%) were by one genus of scorpion Lychas spp. Seventy one percent of stings occurred between 6pm and 8am and 82 (86%) occurred indoors. Sixty percent of stings occurred on distal limbs. The median duration of effects was 6 h (interquartile range (IQR): 1-24 h). Immediate localised pain occurred in all cases and was severe in 76 cases (80%). Other local effects included red mark/redness (66%), tenderness (35%), numbness (12%) and paraesthesia (11%). Minor systemic effects (nausea, headache and malaise) occurred in 11% of cases. There were no deaths or major systemic envenoming. Less severe effects were observed for the larger Urodacus species, compared to Lychas spp. Scorpion stings in Australia do not appear to cause severe or life-threatening effects, even in children. This differs from other parts of the world, where severe envenoming is reported. The major clinical effect is severe pain, consistent with other scorpion stings. Most stings occurred indoors and at night. PMID- 12782089 TI - Molecular cloning and characterization of a platelet glycoprotein Ib-binding protein from the venom of Trimeresurus stejnegeri. AB - A platelet glycoprotein Ib-binding protein, termed TSV-GPIb-BP, was isolated from the venom of Trimeresurus stejnegeri. On SDS-polyacrylamide gel electrophoresis, TSV-GPIb-BP showed a single band with an apparent molecular weight of 28,000 and two distinct bands with apparent molecular weights of 16,000 and 15,000 under non reducing and reducing conditions, respectively. cDNA clones containing the coding sequences for both TSV-GPIb-BP subunits were isolated and sequenced. The deduced amino acid sequences of TSV-GPIb-BP subunits were confirmed by N-terminal protein sequencing and trypsin-digested peptide mass fingerprinting. Interestingly, the alpha subunit of TSV-GPIb-BP is identical to that of alboaggregin-B, and the sequence identity of their beta subunits is 94.3%. TSV-GPIb-BP inhibited ristocetin-induced human platelet agglutination in platelet-rich plasma under lower dosages (<5 microg/ml). On the other hand, it directly aggregated washed human platelets in the absence of additional Ca2+ or any other cofactors under higher dosages (>5 microg/ml). This platelet aggregation activity was dose dependently inhibited by specific GPIbalpha antibodies, but not by those antibodies against platelet GPIa, GPIIa, GPIIb and GPIIIa. PMID- 12782090 TI - Occurrence of 11-oxotetrodotoxin in the red-spotted newt, Notophthalmus viridescens, and further studies on the levels of tetrodotoxin and its analogues in the newt's efts. AB - An analogue of tetrodotoxin (TTX), 11-oxoTTX, was semi-purified from the red spotted newt, Notophthalmus viridescens, and identified by ESI-MS and 1H NMR spectra. The levels of TTX, 11-oxoTTX and 6-epiTTX in early stage of development (efts) of this newt were investigated by a post-column fluorescent-HPLC system, and compared with those of adult newts. The level of 11-oxoTTX in both of adults and efts were remarkably high, almost close to those of TTX, while 6-epiTTX was a minor component in both stages. The level of 6-epiTTX in efts (1.8+/-1.3 microg/g, SD, n=10) was significantly larger than that in adults (0.51+/-0.26 microg/g, SD, n=12) (p<0.05), while no significant difference was observed in the levels of TTX and 11-oxoTTX between efts (13+/-7.4 and 9.1+/-5.6 microg/g) and adult newts (16+/-6.3 and 13+/-6.2 microg/g, respectively) (p>0.05). PMID- 12782091 TI - Contribution of crotoxin for the inhibitory effect of Crotalus durissus terrificus snake venom on macrophage function. AB - Previous work of our group demonstrated that Crotalus durissus terrificus venom has a dual effect on macrophage function: it inhibits spreading and phagocytosis and stimulates hydrogen peroxide and nitric oxide production, antimicrobial activity and glucose and glutamine metabolism of these cells. Crotalid venom also induces analgesia and this effect is mediated by opioid receptors. The involvement of opioidergic mechanism and the determination of the active component responsible for the inhibitory effect of crotalid venom on macrophage function were investigated. The venom reduced the spreading and phagocytic activities of peritoneal macrophages. This effect was observed in vitro, 2 h after incubation of resident peritoneal macrophages with the venom, and in vivo, 2 h after subcutaneous injection of the venom. The inhibition of phagocytosis was not modified by naloxone, an antagonist of opioid receptors. Venom neutralization with crotalid antivenom abolished the inhibitory effect of the venom, indicating that venom toxins are involved in this effect. Crotoxin, the main toxin of crotalid venom, s.c. injected to rats or added to the medium of peritoneal cell incubation, inhibited macrophage function in a similar manner to that observed for crude venom. The present results suggest that crotoxin causes a direct inhibition of macrophage spreading and phagocytic activities and may contribute to the inhibitory effect of crotalid venom on macrophage function. PMID- 12782092 TI - Molecular cloning of a gamma-phospholipase A2 inhibitor from Lachesis muta muta (the bushmaster snake). AB - Several endogenous phospholipase A(2) inhibitors (PLIs) have been purified from the blood plasma of a number of snake species and are classified into three classes (alpha, beta and gamma) according to their structure and specificity. In the present study we have cloned transcripts of a protein homologous to CNF, a gammaPLI present in Crotalus durissus terrificus plasma, that is encoded in the liver of Lachesis muta muta (the bushmaster snake), a species evolutionarily related to Crotalus. The cDNA sequences code for two isoforms of a 200-residue protein including a 19-residue signal peptide followed by 181 amino acid residues in the mature form and a putative N-linked carbohydrate site. The deduced primary structures and some properties of those new proteins were compared to those of CNF. Multiple alignment was performed with the aminoacid sequences of all the gammaPLIs described so far and this used in the construction of a phylogenetic tree. PMID- 12782093 TI - Type B brevetoxins show tissue selectivity for voltage-gated sodium channels: comparison of brain, skeletal muscle and cardiac sodium channels. AB - Brevetoxins and ciguatoxins are two classes of phycotoxins which exert their toxic effect by binding to site-5 of voltage-gated sodium channels. Sodium channels, a family of at least 10 structurally different proteins, are responsible for the rising phase of the action potential in membranes of neuronal, cardiac and muscular excitable cells. This work is a comparative study of the binding properties and the cytotoxic effects of ciguatoxins and brevetoxins on human embryonic cells (HEK) stably expressing either the skeletal muscle (Na(v)1.4), or the cardiac (Na(v)1.5) sodium channel alpha-subunit isoforms. We report that type A (PbTx-1) and type B (PbTx-3 and PbTx-2) brevetoxins as well as ciguatoxins target both cardiac and muscle channels; type B brevetoxins show isoform selectivity, presenting a lower affinity for the heart than the skeletal muscle channel. The lower selectivity of type B brevetoxins for heart sodium channels may result from a more rigid backbone structure than is found in type A brevetoxins and ciguatoxins. PMID- 12782094 TI - Preliminary in vitro studies on the Marsypianthes chamaedrys (boia-caa) extracts at fibrinoclotting induced by snake venoms. AB - The extract of Marsypianthes chamaedrys, a plant used against snakebites, in the present study was shown to inhibit fibrinoclotting induced by several Brazilian snake venoms or thrombin. These data indicate that this extract affected thrombin like enzymes. In this first report we determine some features of the components present in the extract regarding the antifibrinoclotting action. Our results show that active components responsible for those effects are thermo-resistant and are concentrated in the methanolic fraction. PMID- 12782095 TI - Tremorgenic syndrome in goats caused by Ipomoea asarifolia in Northeastern Brazil. AB - Green leaves of Ipomoea asarifolia were dosed to 10 goats. Nine goats ingesting 5 37 g/kg bw daily had clinical signs in 4-38 days. One goat ingesting 2.5 g/kg bw daily during 125 days and two control goats had no clinical signs. Clinical signs were characteristic for a tremorgenic syndrome. Five goats recovered in 4-9 days after the withdrawal of the plant. Two goats died spontaneously and three were euthanased for histologic and ultrastructural studies. No significant lesions were observed at necropsies or on the histologic and ultrastructural studies. Samples of the plant analyzed for enzymatic inhibitors were negative for calystegines and contained an almost undetectable amount of swainsonine (less than 0.001%). It is concluded that I. asarifolia causes a tremorgenic syndrome due to an unknown tremorgenic phytotoxins or mycotoxins. PMID- 12782096 TI - Bibliography of toxinology. PMID- 12782098 TI - Use of microarray technologies in toxicology research. AB - Microarray technology provides a unique tool for the determination of gene expression at the level of messenger RNA (mRNA). The simultaneous measurement of the entire human genome (thousands of genes) will facilitate the uncovering of specific gene expression patterns that are associated with disease. One important application of microarray technology, within the context of neurotoxicological studies, is its use as a screening tool for the identification of molecular mechanisms of toxicity. Such approaches enable researchers to identify those genes and their products (either single or whole pathways) that are involved in conferring resistance or sensitivity to toxic substances. This review addresses: (1) the potential uses of array data; (2) the various array platforms, highlighting both their advantages and disadvantages; (3) insights into data analysis and presentation strategies; and (4) concrete examples of DNA array studies in neurotoxicological research. PMID- 12782099 TI - Cyanide enhancement of dopamine-induced apoptosis in mesencephalic cells involves mitochondrial dysfunction and oxidative stress. AB - Dopamine (DA)-induced neurotoxicity is potentiated when cellular metabolism is compromised. Since cyanide is a neurotoxin that produces mitochondrial dysfunction and stimulates intracellular generation of reactive oxygen species (ROS), KCN was used to study DA-induced apoptosis in primary cultured mesencephalon cells. Treatment of neurons with DA (300 microM) for 24h produced apoptosis as determined by TUNEL staining, DNA fragmentation and increased caspase activity. Pretreatment with KCN (100 microM) 30min prior to DA increased the number of cells undergoing apoptosis. When added to the cells alone, this concentration of KCN did not induce apoptosis. DA stimulated intracellular generation of ROS, and treatment with KCN enhanced ROS generation. Treatment of cells with glutathione or uric acid (antioxidants/scavengers) attenuated both the increase in ROS generation and the apoptosis, demonstrating that ROS are initiators of the cytotoxicity. Studies on the sequence of events mediating the response showed that DA-induced depolarization of the mitochondrial membrane was dependent on ROS generation and KCN enhanced this action of DA. Following changes in mitochondrial membrane potential, cytochrome c was released from mitochondria, leading to caspase activation and eventually cell death. These results demonstrate that oxidative stress and mitochondrial dysfunction are initiators of DA-induced apoptosis. Subsequent cytochrome c release activates the caspase effector component of apoptosis. Cyanide potentiates the neurotoxicity of DA by enhancing the generation of ROS and impairing mitochondrial function. PMID- 12782100 TI - Dentate gyrus: alterations that occur with hippocampal injury. AB - Injury to the brain usually manifests not in a diffuse uniform manner but rather with selective sites of damage indicative of differential vulnerability. This question of neuronal susceptibility has been one of major interest both in disease processes as well as damage induced by environmental factors. For experimental examination, brain structures with obvious neuronal subpopulations and organization such as the cerebellum and the hippocampus have offered the most promise. In the hippocampus distinct neuronal populations exist that demonstrate differential vulnerability to various forms of insult including ischemia, excitotoxicity, and environmental factors. The more recent data regarding the presence of neuronal progenitor cells in the subgranular zone of the dentate offers the opportunity to expand such experimental examination to the process of injury-induced neurogenesis. Thus, more recent studies have expanded the examination of the hippocampus to include models of damage to the dentate neurons in addition to the highly vulnerable pyramidal neurons. A number of these models are presented for both human disease and experimental animal conditions. Examination of the responses between these distinct cell populations offers the potential for understanding factors that are critical in neuronal death and survival. PMID- 12782101 TI - Pharmacological agents, hippocampal EEG, and anticonvulsant effects on soman induced seizures in rats. AB - Changes in the hippocampal theta rhythm were used as a model in which anticonvulsant drugs may be screened for their potential to antagonize soman induced (1xLD(50)) seizures. The zinc chelator, ethylenediaminetetra acetic acid (EDTA) (300mg/kg), and the NMDA receptor antagonist, HA-966 (60mg/kg), both disrupted the theta rhythm, but did not antagonize soman-induced seizures, neither separately, nor in combination. The anticholinergic and antiglutamatergic procyclidine (6mg/kg) did not influence the theta activity. The GABAergic agonists, diazepam (10mg/kg) and pentobarbital (30mg/kg), both reduced the theta frequency. Procyclidine, diazepam, and pentobarbital did not stop soman-induced seizures when administered separately, but both convulsions and seizure activity terminated when these agents were given together, and the rats slept through the critical convulsion period. This triple therapy was 100% effective, when administered 30-40min following onset of convulsions, and the rats displayed apparently normal behavior the next day. A screening model of potential anticonvulsants cannot be based on alterations in hippocampal EEG activity. Procyclidine, diazepam, and pentobarbital in combination disrupted the theta rhythm like the combination of EDTA and HA-966, but the latter combination did not have anticonvulsant effect. It is concluded that a triple regimen consisting of procyclidine, diazepam, and pentobarbital can effectively terminate soman induced seizures that have lasted 30min or more. PMID- 12782102 TI - Methylmercury induces apoptosis in cultured rat dorsal root ganglion neurons. AB - Methylmercury is known to have devastating effects on the mammalian nervous system. In order to characterize the dose dependence of methylmercury-induced neurotoxicity, we first studied neurite outgrowth from rat dorsal root ganglia explants. In this model, methylmercury inhibited neurite outgrowth with a TD(50) of approximately 0.5 microM. We then used this relationship to optimize dosing for subsequent transcriptional profiling analyses in two independent neuronal model systems: dissociated sensory neurons and PC12 cells. As seen in previous studies, the expression of a number of genes associated with oxidative stress was altered following a 6h challenge with 1 microM methylmercury. When PC12 cells were subjected to a longer exposure (24h), a relative increase was noted in the representation of genes associated with cell cycling and apoptosis. To confirm the presence of apoptosis in cultured neurons, we then applied TUNEL staining and bis-benzimide staining techniques to primary cultures of dissociated sensory neurons. After 24h, 1 microM methylmercury increased both DNA end-labeling (P<0.01) and nuclear fragmentation (P<0.02). The latter effect appeared to be dose-dependent. PMID- 12782104 TI - Regional variation in the activation threshold for 1,3-DNB-induced mitochondrial permeability transition in brainstem and cortical astrocytes. AB - 1,3-Dinitrobenzene (DNB) produces edematous, glio-vascular lesions in brainstem nuclei with high energy demands. Astrocytes in vulnerable brainstem nuclei appear to be an early and selective target of DNB and other nitroaromatic compounds, though the molecular basis of this susceptibility is poorly understood. It has been postulated that mitochondria are a principal target of DNB in sensitive cell types [Neuropathol. Appl. Neurobiol. 13 (5) (1987) 371], where redox-cycling of DNB increases levels of reactive oxygen species and disrupts cellular energy metabolism. The present study investigates the role of regional differences in activation of the mitochondrial permeability transition pore (mtPTP) by DNB in brainstem and cortical astrocytes and examines the expression of Bcl-2 proteins as potential regulators of mtPTP function. Neonatal rat astrocytes were cultured from both DNB-sensitive (brainstem) and insensitive (cortex) brain regions and evaluated for DNB-induced alterations in cell morphology and mitochondrial function. Exposure to DNB resulted in rapid changes in the morphology of brainstem astrocytes consistent with loss of ion homeostasis and initiation of necrotic cell death. These changes were not observed in cortical astrocytes at corresponding concentrations of DNB and were prevented in brainstem astrocytes by the mtPTP inhibitor, bongkrekic acid, suggesting that mitochondrial dysfunction is involved in DNB-induced morphological changes in brainstem astrocytes. Mitochondrial depolarization in brainstem astrocytes was observed at DNB concentrations as low as 10 microM, whereas no loss of mitochondrial membrane potential (DeltaPsi(mt)) occurred in cortical astrocytes at less than 100 microM DNB. DNB-induced loss of DeltaPsi(mt) followed apparent first-order kinetics, with EC(50)-values for half-maximal rates of mitochondrial depolarization of approximately 23 and approximately 290 microM in brainstem cortical astrocytes, respectively. DNB-induced mitochondrial depolarization was prevented by pretreatment with bongkrekic acid, indicating that loss of DeltaPsi(mt) was mediated by activation of the mtPTP. Inhibition of succinate dehydrogenase (SDH) activity occurred in astrocytes from both brain regions exposed to DNB and was blocked in brainstem, but not cortical, astrocytes by bongkrekic acid. Constitutive expression of Bcl-X(L) was high in cortical tissue and astrocytes, whereas Bax expression was low. However, Bax was highly expressed in brainstem tissue and astrocytes and Bcl-X(L) expression was markedly lower. The expression of Bcl-2 was similar in both brain regions. These data suggest that the selective vulnerability of brainstem astrocytes to DNB is due to a lower threshold for activation of the mtPTP that is be mediated, in part, by distinct expression patterns of Bcl-2 proteins rather than by intrinsic differences in susceptibility of the electron transport chain. PMID- 12782103 TI - Plasma levels of parent compound and metabolites after doses of either d fenfluramine or d-3,4-methylenedioxymethamphetamine (MDMA) that produce long-term serotonergic alterations. AB - Plasma levels of parent compounds and metabolites were determined in adult rhesus monkeys after doses of either 5mg/kg d-fenfluramine (FEN) or 10mg/kg d-3, 4 methylenedioxymethamphetamine (MDMA) i.m. twice daily for four consecutive days. These treatment regimens have been previously shown to produce long-term serotonin (5-HT) depletions. Peak plasma levels of 2.0+/-0.4 microM FEN were reached within 40min after the first dose of FEN, and then declined rapidly, while peak plasma levels (0.4+/-0.1 microM) of the metabolite norfenfluramine (NFEN) were not reached until 6h after dosing. After the seventh (next to last) dose of FEN, peak plasma levels of FEN were 35% greater than after the first dose while peak NFEN-levels were 500% greater. The t(1/2) for FEN was 2.6+/-0.3h after the first dose and 3.2+/-0.2h after the seventh. The estimated t(1/2) for NFEN was more than 37.6+/-20.5h. Peak plasma levels of 9.5+/-2.5 microM MDMA were reached within 20min after the first dose of MDMA, and then declined rapidly, while peak plasma levels (0.9+/-0.2 microM) of the metabolite 3,4 methylenedioxyamphetamine (MDA) were not reached until 3-6h after dosing. After the seventh (next to last) dose of MDMA, peak plasma levels of MDMA were 30% greater than the first dose while peak MDA levels were elevated over 200%. The t(1/2) for MDMA was 2.8+/-0.4h after the first and 3.9+/-1.1h after the seventh dose. The estimated t(1/2) for MDA was about 8.3+/-1.0h. Variability in plasma levels of MDMA and MDA between subjects was much greater than that for FEN and NFEN. This variability in MDMA and MDA exposure levels may have lead to variability in the subsequent disruption of some behaviors seen in these same subjects. There were 80% reductions in the plasma membrane-associated 5-HT transporters 6 months after either the FEN or MDMA dosing regimen indicating that both treatments produced long-term serotonergic effects. PMID- 12782105 TI - 1,3-Dinitrobenzene inhibits mitochondrial complex II in rat and mouse brainstem and cortical astrocytes. AB - 1,3-Dinitrobenzene (DNB) produces edematous, glio-vascular lesions that are initially confined to brainstem nuclei with high energy requirements in rats and mice. Perturbation of energy producing processes in the cell is known to induce formation of the mitochondrial permeability transition pore (mtPTP) complex. Selective vulnerability of brainstem astrocytes to DNB is mediated by a 10-fold lower threshold for opening of the cyclosporin A-inhibitable mitochondrial permeability transition (MPT) pore than their cortical counterparts. Other nitrocompounds, such as 3-nitropropionic acid, selectively interfere with regional energy metabolism, including mitochondrial succinate dehydrogenase activity. However, the link between DNB-induced onset of the MPT and disruption of energy producing processes in the astrocyte remains unclear. The effects of DNB on succinate dehydrogenase activity were evaluated in cultured neonatal rat and mouse brainstem and cortical astrocytes. Both histochemical and spectrophotometric assays confirmed significant temporal inhibition of SDH activity in brainstem and cortical astrocytes 0.5, 2 and 5h following exposure to 100 microM DNB in vitro. Although DNB-induced inhibition of SDH was significantly decreased by CsA pretreatment in brainstem astrocytes after 0.5 and 2h and with a second pore inhibitor, bongkrekic acid (BKA) after 5h, both inhibitors failed to reduce inhibition of SDH activity in cortical astrocytes. These data suggest that DNB-induced inhibition of SDH may be independent of differential regional activation of the mtPTP complex in astrocytes and that an unidentified cyclosporin A-inhibitable factor mediates DNB-induced loss of SDH function. PMID- 12782106 TI - 1-Benzyl-1,2,3,4-tetrahydroisoquinoline (1BnTIQ), an endogenous neurotoxin, induces dopaminergic cell death through apoptosis. AB - Endogenous MPTP-like neurotoxins such as 1-benzyl-1,2,3,4-tetrahydroisoquinoline (1BnTIQ) have been suspected in the etiology of Parkinson's disease (PD). 1BnTIQ was found in a concentration three times higher in cerebrospinal fluid of PD brains than control subjects [J. Neurochem. 65 (6) (1995) 2633]. In the present study, we have evaluated the mechanisms of 1BnTIQ toxicity in human dopaminergic SH-SY5Y cells and tested the neuroprotective action of SKF-38393, a dopamine receptor (D(1)) agonist. 1BnTIQ dose dependently decreased cell viability in dopaminergic SH-SY5Y cells and the extent of cell death was more pronounced when compared to MPP(+). Similar to MPP(+), 1BnTIQ significantly decreased [3H]dopamine uptake. 1BnTIQ significantly increased lipid peroxidation, Bax expression, and active caspase-3 formation. Furthermore, it decreased the expression of Bcl-xL, an anti-apoptotic protein, in these cells. SKF-38393, a dopamine receptor (D(1)) agonist (1 and 10 microM) completely prevented the cell death and significantly increased cell viability. These results strongly suggest that 1BnTIQ induces dopaminergic cell death by apoptosis and dopamine receptor agonists may be useful neuroprotective agents against 1BnTIQ toxicity. PMID- 12782107 TI - Thiram and ziram stimulate non-selective cation channel and induce apoptosis in PC12 cells. AB - The neurotoxicity of dithiocarbamates has been previously reported, however, the detailed mechanism underlying the neurotoxicity is still not fully understood. Among the dithiocarbamates, we investigated thiram and ziram in a neuronal-like pheochromocytoma (PC12) cells. Thiram and ziram strongly induced cell death in both dose- and time-dependent manners with the LC(50) of 0.3 and 2 microM, respectively. The cell death showed typical apoptotic features, such as DNA fragmentation and an increase of subdiploidy nuclei. Interestingly, both thiram and ziram induced rapid and sustained increases of intracellular Ca(2+) in PC12 cells, which were almost completely blocked by flufenamic acid (FFA), an inhibitor of non-selective cation channel. BAPTA-AM, an intracellular Ca(2+) chelator, inhibited the thiram- and ziram-induced apoptotic cell death. These results suggest that thiram and ziram induce apoptotic neuronal cell death by Ca(2+) influx through non-selective cation channels. The present study may provide a clue for understanding the mechanism of neurotoxicity of thiram and ziram. PMID- 12782108 TI - Behavioral effects of chronic exposure to low levels of lead in Drosophila melanogaster. AB - Through human activity lead has become a serious environmental neurotoxin, known to affect activity levels, attention and both sensory and cognitive function in children. Study of lead would be facilitated by having a model system that could be manipulated easily and quickly. We find Drosophila melanogaster ideal as such, and we have been studying effects of lead on courtship, fecundity and locomotor activity. We raised Canton-S flies from eggs to adult day 6-7 on medium made with lead acetate solution (2-100 microgram/g), or with distilled water, and we measured adult body lead burdens by means of Inductively Coupled Plasma Mass Spectrometry (ICP-MS). To measure courtship effectiveness, five virgin females and five virgin males were transferred into an empty vial and the number of females that mated within 20min was recorded. To measure fecundity, all adult offspring from eggs produced by one female within 12 days of mating were counted. To measure locomotor activity, individual flies were transferred to a grid labeled petri dish and the number of lines crossed in 30s was counted. The number of females mating within 20min was increased significantly by exposure to 2 or 8 microgram/g lead, and was decreased significantly by exposure to 20 or 50 microgram/g lead. Fecundity was increased significantly by exposure to 2 microgram/g lead, but was unaffected by exposure to 20 microgram/g lead. Locomotor activity was consistently higher for males than for females, and was significantly reduced only by exposure to 50 microgram/g lead, and then only for males. We thus defined for Drosophila a lowest observable effect level (LOEL) of 2 microgram/g lead, which is considerably lower than the doses shown previously to affect this animal. The dose-response curve was biphasic for the number of females mating within 20min, an example of hormesis, a non-linear response that has been reported for low levels of stressors as diverse as pollutants and radiation. We hope from further studies with Drosophila to understand better how lead affects the developing nervous system, and thus ultimately its effects on children. PMID- 12782109 TI - The effect of tertiary butylhydroperoxide and nicotinamide on human cortical neurons. AB - It is well known that the generation of oxygen radicals can cause neuronal death by both apoptosis and necrosis, which may lead to the onset of neurodegenerative diseases. In previous in vivo studies, nicotinamide was found to prevent both DNA fragmentation and apoptosis that were induced by free radical generating toxins like tertiary butylhydroperoxide (t-BuOOH). Nicotinamide is a precursor for NAD and is an inhibitor of the enzyme poly(ADP-ribose) polymerase (PARP). However, the effect of nicotinamide on the regulation of pro- and anti-apoptotic proteins in neurons is not clear. In our study, the human cortical neuronal cell line HCN1 A has been used to determine the mechanism of action of nicotinamide at the cellular level. Cell viability studies showed that t-BuOOH treatment (both 100 microM and 1mM) caused significant cell death at 24 and 48h compared to control cells. Pretreatment with 1mM nicotinamide before t-BuOOH administration caused significant reduction in cell death. Moreover, the morphology of HCN1-A cells that were treated with both nicotinamide and t-BuOOH appeared to be closer to control cells when compared to HCN1-A cells treated with only t-BuOOH. Also, t BuOOH treatment caused an elevation in the levels of the pro-apoptotic proteins p53 and p21/WAF-1 and a reduction in the levels of the anti-apoptotic protein bcl 2 compared to their levels in control HCN1-A cells, while pretreatment with nicotinamide reduced p53 and p21/WAF-1 levels even in the presence of t-BuOOH. However nicotinamide did not seem to alter bcl-2 levels. These results indicate that nicotinamide treatment can protect human neuronal cells from the toxic effects of t-BuOOH. PMID- 12782110 TI - Polybrominated diphenyl ethers: neurobehavioral effects following developmental exposure. AB - Polybrominated diphenyl ethers (PBDEs), a class of widely used flame retardants, are becoming widespread environmental pollutants, as indicated by studies on sentinel animal species, as well as humans. Of particular concern are the reported increasingly high levels of PBDEs in human milk, as should be given that almost no information is available on their potential effects on developing organisms. In order to address this issue, studies have been conducted in mice and rats to assess the potential neurotoxic effects of perinatal exposure to PBDEs (congeners 47, 99, 153 and the penta-BDE mixture DE-71). Characteristic endpoints of PBDE neurotoxicity are, among others, endocrine disruption (e.g. decreased thyroid hormone levels), alteration in cholinergic system activity (behavioral hyporesponsivity to nicotine challenge), as well as alterations of several behavioral parameters. In particular, the main hallmark of PBDE neurotoxicity is a marked hyperactivity at adulthood. Furthermore, a deficit in learning and memory processes has been found at adulthood in neonatally exposed animals. Some of neurotoxic effects of PBDEs are comparable to those of polychlorinated biphenyls (PCBs), though the latter class of compounds seems to exert a stronger toxic effect. Available information on PBDE neurotoxicity obtained from animal studies and the possibility of neonatal exposure to PBDEs via the mother's milk suggest that these compounds may represent a potential risk for neurobehavioral development in humans. PMID- 12782111 TI - Electrostatic charge activates inflammatory vanilloid (VR1) receptors. AB - The pathophysiology of neurogenic inflammation culminates in the overt symptoms of tissue inflammation through a series of events which are initiated by the activation of vanilloid receptors (VR1). This study was designed to test the hypothesis that a sufficiently negative, electrostatic charge carried on a particulate matter (PM) particle, could acquire a cloud of protons sufficient to activate proton-sensitive VR1 receptors and acid-sensitive ionic channels (ASICs) pathways. For this, nanometer-sized, synthetic polystyrene micells (SPM) or those charged with chemical groups (e.g. diamino, carboxyl) were used. These chemical groups imparted either a net positive (i.e. diamino) or negative (i.e. carboxyl) charge on the SPM when suspended in a neutral ionic medium. The zeta potential, a measure of the SPM's electronegativity, was taken in both cell culture nutrient medium and in ultraviolet light-distilled water (UV-DW). In both vehicles, the rank order of electronegativity (most to least negative) was carboxyl > polystyrene > diamino-SPM. Individual types of SPM were exposed to human, immortalized bronchial-tracheal epithelial cells (i.e. BEAS-2B) and endpoints of biological activation (i.e. membrane depolarization, increases in intracellular calcium (i.e. [Ca(2+)](i)) levels, IL-6 release) were measured. Cells loaded with a fluorescent probe for membrane depolarization (3,3'-dihexyloxacarbocyanine iodide, DiOC-6-3) showed a positive reaction when exposed to carboxyl-SPM but not to diamino-SPM. BEAS-2B cells exposed to carboxyl-SPM responded with significant increases in [Ca(2+)](i), and IL-6 release relative to uncharged SPM or diamino SPM. This IL-6 release could be reduced by pretreatment with antagonists to the VR1 receptor (i.e. capsazepine) or to acid-sensitive ionc channels (i.e. amiloride). Although both diamino and carboxyl-SPM groups stimulated increases in IL-6 transcript, only the more electronegatively charged carboxyl-SPM stimulated mRNA-VR1 receptor. These data suggest that measurable inflammatory changes can be stimulated in human epithelial target cells by the electrostatic charge carried on an inert particle. Further, these changes appear to be mediated through acid sensitive VR1 receptors and ASICs. PMID- 12782112 TI - The topology of the regulatory interactions predicts the expression pattern of the segment polarity genes in Drosophila melanogaster. AB - Expression of the Drosophila segment polarity genes is initiated by a pre-pattern of pair-rule gene products and maintained by a network of regulatory interactions throughout several stages of embryonic development. Analysis of a model of gene interactions based on differential equations showed that wild-type expression patterns of these genes can be obtained for a wide range of kinetic parameters, which suggests that the steady states are determined by the topology of the network and the type of regulatory interactions between components, not the detailed form of the rate laws. To investigate this, we propose and analyse a Boolean model of this network which is based on a binary ON/OFF representation of mRNA and protein levels, and in which the interactions are formulated as logical functions. In this model the spatial and temporal patterns of gene expression are determined by the topology of the network and whether components are present or absent, rather than the absolute levels of the mRNAs and proteins and the functional details of their interactions. The model is able to reproduce the wild type gene expression patterns, as well as the ectopic expression patterns observed in overexpression experiments and various mutants. Furthermore, we compute explicitly all steady states of the network and identify the basin of attraction of each steady state. The model gives important insights into the functioning of the segment polarity gene network, such as the crucial role of the wingless and sloppy paired genes, and the network's ability to correct errors in the pre-pattern. PMID- 12782113 TI - Interpreting time-series analyses for continuous-time biological models--measles as a case study. AB - An increasing number of recent studies involve the fitting of mechanistic models to ecological time-series. In some cases, it is necessary for these models to be discrete-time approximations of continuous-time processes. We test the validity of discretization in the case of measles, where time-series models have recently been developed to estimate ecological parameters directly from data. We find that a non-homogeneous contact function is necessary to capture the host-parasite interaction in a discrete-time model, even in the absence of heterogeneities due to spatial or age structure. We derive a mathematical relationship describing the expected departure from mass-action transmission in terms of the epidemiological parameters in the model, and identify conditions under which the discretization process may fail. PMID- 12782114 TI - Length analyses of Drosophila odorant receptors. AB - Odorant receptors comprise a unique family of G-protein-coupled seven transmembrane receptors both in mammals and insects. In the fruit fly Drosophila melanogaster, all 61 candidate odorant receptor genes have been identified based on the complete genome sequence, and their expression patterns have been examined. A given odorant receptor is expressed in the antenna or maxillary palp, or not expressed at all. Here we have applied a set of statistical analyses to the length of the extra- and intracellular loops and terminals (LTs) of Drosophila odorant receptors to examine possible inter- and intramolecular relations at the population level. We have first provided some useful statistical information such as mean length values and length histograms to depict a general nature of Drosophila odorant receptors at the population level, after focusing on discrepancy on assigning transmembrane domains between researchers. In a preferable transmembrane assignment, all extracellular LTs, especially the second extracellular loops, were relatively large in length, suggesting their functional significance. Somewhat surprisingly, principle component analysis (PCA) indicated that the maxillary palp receptors were almost as diverse as the antenna receptors despite their much smaller population size. PCA together with histograms also revealed that receptors with an abnormal length configuration tended not to be expressed, suggesting that LT length deviations are related to transcriptional silencing of odorant receptor genes. Rank transformation tests pointed out possible LTs that could have different length between differently expressed receptors at the population level. Taken together, length analyses provide us with a general picture, i.e. "length configuration," of Drosophila odorant receptors at the population level that could point out putatively important functional sites for experimental studies. PMID- 12782115 TI - Catastrophic desert formation in Daisyworld. AB - Feedback between life and its environment is ubiquitous but the strength of coupling and its global implications remain hotly debated. Abrupt changes in the abundance of life for small changes in forcing provide one indicator of regulation, for example, when vegetation-climate feedback collapses in the formation of a desert. Here we use a two-dimensional "Daisyworld" model with curvature to show that catastrophic collapse of life under gradual forcing provides a testable indicator of environmental feedback. When solar luminosity increases to a critical value, a desert forms across a wide band of the planet. The scale of collapse depends on the strength of feedback. The efficiency of temperature regulation is limited by mutation rate in an analogous manner to the limitation of adaptive fitness in evolutionary theories. The final state of the system emerging from single-site rules can be described by two global quantities: optimization of temperature regulation and maximization of diversity, which are mathematically analogous to energy and entropy in thermodynamics. PMID- 12782116 TI - Centers of complex networks. AB - The central vertices in complex networks are of particular interest because they might play the role of organizational hubs. Here, we consider three different geometric centrality measures, excentricity, status, and centroid value, that were originally used in the context of resource placement problems. We show that these quantities lead to useful descriptions of the centers of biological networks which often, but not always, correlate with a purely local notion of centrality such as the vertex degree. We introduce the notion of local centers as local optima of a centrality value "landscape" on a network and discuss briefly their role. PMID- 12782117 TI - Biochemical and genomic regulation of the trehalose cycle in yeast: review of observations and canonical model analysis. AB - The physiological hallmark of heat-shock response in yeast is a rapid, enormous increase in the concentration of trehalose. Normally found in growing yeast cells and other organisms only as traces, trehalose becomes a crucial protector of proteins and membranes against a variety of stresses, including heat, cold, starvation, desiccation, osmotic or oxidative stress, and exposure to toxicants. Trehalose is produced from glucose 6-phosphate and uridine diphosphate glucose in a two-step process, and recycled to glucose by trehalases. Even though the trehalose cycle consists of only a few metabolites and enzymatic steps, its regulatory structure and operation are surprisingly complex. The article begins with a review of experimental observations on the regulation of the trehalose cycle in yeast and proposes a canonical model for its analysis. The first part of this analysis demonstrates the benefits of the various regulatory features by means of controlled comparisons with models of otherwise equivalent pathways lacking these features. The second part elucidates the significance of the expression pattern of the trehalose cycle genes in response to heat shock. Interestingly, the genes contributing to trehalose formation are up-regulated to very different degrees, and even the trehalose degrading trehalases show drastically increased activity during heat-shock response. Again using the method of controlled comparisons, the model provides rationale for the observed pattern of gene expression and reveals benefits of the counterintuitive trehalase up regulation. PMID- 12782118 TI - Optimal diving under the risk of predation. AB - Many air-breathing aquatic foragers may be killed by aerial or subsurface predators while recovering oxygen at the surface; yet the influence of predation risk on time allocation during dive cycles is little known in spite of numerous studies on optimal diving. We modeled diving behavior under the risk of predation at the surface. The relationship between time spent at the surface and the risk of death is predicted to influence the optimal surface interval, regardless of whether foragers accumulate energy at a constant rate while at the food patch, deplete food resources over the course of the dive, or must search for food during the dive. When instantaneous predation risk during a single surface interval decreases with time spent at the surface, a diver should increase its surface interval relative to that which maximizes energy intake, thereby increasing dive durations and reducing the number of surfacings per foraging bout. When instantaneous risk over a single surface interval does not change or increases with increasing time at the surface, divers should decrease their surface interval (and consequently their dive duration) relative to that which maximizes energy intake resulting in more dives per foraging bout. The fitness consequences of selecting a suboptimal surface interval vary with the risk function and the way divers harvest energy when at depth. Finally, predation risk during surface intervals should have important consequences for habitat selection and other aspects of the behavioral ecology of air-breathing aquatic organisms. PMID- 12782119 TI - Metabotropic receptor activation, desensitization and sequestration-I: modelling calcium and inositol 1,4,5-trisphosphate dynamics following receptor activation. AB - A mathematical account is given of the processes governing the time courses of calcium ions (Ca2+), inositol 1,4,5-trisphosphate (IP(3)) and phosphatidylinositol 4,5-bisphosphate (PIP(2)) in single cells following the application of external agonist to metabotropic receptors. A model is constructed that incorporates the regulation of metabotropic receptor activity, the G-protein cascade and the Ca2+ dynamics in the cytosol. It is subsequently used to reproduce observations on the extent of desensitization and sequestration of the P(2)Y(2) receptor following its activation by uridine triphosphate (UTP). The theory predicts the dependence on agonist concentration of the change in the number of receptors in the membrane as well as the time course of disappearance of receptors from the plasmalemma, upon exposure to agonist. In addition, the extent of activation and desensitization of the receptor, using the calcium transients in cells initiated by exposure to agonist, is also predicted. Model predictions show the significance of membrane PIP(2) depletion and resupply on the time course of IP(3) and Ca2+ levels. Results of the modelling also reveal the importance of receptor recycling and PIP(2) resupply for maintaining Ca2+ and IP(3) levels during sustained application of agonist. PMID- 12782120 TI - Metabotropic receptor activation, desensitization and sequestration-II: modelling the dynamics of the pleckstrin homology domain. AB - Recent observations have been made regarding the generation of inositol 1,4,5 trisphosphate (IP(3)), using chimeras of green fluorescent protein and the pleckstrin homology domain of phospholipase C-delta. In this paper a model is presented giving the quantitative relations between the green fluorescent protein pleckstrin homology domain (GFP-PHD) construct and membrane phosphatidylinositol 4,5-bisphosphate (PIP(2)) levels as well as the concentration of IP(3), the product of hydrolysis of PIP(2). The model can correctly reproduce the dependence of cytosolic GFP-PHD fluorescence on IP(3) concentration. This model extends a previous one (Metabotropic receptor activation, desensitization and sequestration I: modelling calcium and inositol 1,4,5-trisphosphate dynamics following receptor activation, in this issue) dealing with the processes governing the production of IP(3) and the subsequent calcium (Ca2+) changes in cells following activation of metabotropic receptors. This model is applied to the case of purinergic P(2)Y(2) receptor activation in Madin-Darby Canine Kidney (MDCK) cells with adenosine triphosphate (ATP) (Science 284 (1999) 1527). It is shown that it can correctly reproduce the dependence of GFP-PHD fluorescence on the concentration of P(2)Y(2) receptor ligand, as well as the temporal changes of GFP-PHD fluorescence following application of ligand. PMID- 12782121 TI - Melanin and HIV in sub-Saharan Africa. AB - HIV is common in sub-Saharan Africa. Sexually transmitted bacterial and fungal infections increase the chance of HIV infection. Melanin can prevent the penetration of skin and mucus membranes by microorganisms, and soluble melanin can inhibit HIV replication. We suggest that melanin may reduce the incidence of HIV infection through venereally acquired skin lesions, thus reducing the risk of sero-conversion and slow the progress to AIDS. Indigenous sub-Saharan peoples are highly melanized, but there is pigment variation between populations. We show that skin reflectance, a negative correlate of melanin, is positively associated with adult rate of HIV in sub-Saharan countries. There is no such relationship in populations outside sub-Saharan Africa. We suggest that melanin concentration in black people may correlate with resistance to HIV infection. PMID- 12782122 TI - Genomic studies of gene expression: regulation of the Wilson disease gene. AB - Bacterial artificial chromosomes (BACs) have many advantages over other large insert cloning vectors and have been used for a variety of genetic applications, including the final contigs of the human genome. We describe the utilization of a BAC construct to study gene regulation in a tissue culture-based system, using a 170-kb clone containing the entire Wilson disease (WND) locus as a model. A second BAC construct that lacked a putative negatively regulating promoter sequence was created. A nonviral method of gene delivery was applied to transfect three human cell lines stably with each construct. Our results show correct WND gene expression from the recombinant locus and quantification revealed significantly increased expression from the clone lacking the negative regulator. Comparison with conventional methods confirms the reliability of the genomic approach for thorough examination of gene expression. This experimental system illustrates the potential of BAC clones in genomic gene expression studies, new gene therapy strategies, and validation of potential molecular targets for drug discovery. PMID- 12782123 TI - Gene expression profile of human lymphoid CEM cells sensitive and resistant to glucocorticoid-evoked apoptosis. AB - Three closely related clones of leukemic lymphoid CEM cells were compared for their gene expression responses to the glucocorticoid dexamethasone (Dex). All three contained receptors for Dex, but only two responded by undergoing apoptosis. After a time of exposure to Dex that ended late in the interval preceding onset of apoptosis, gene microarray analyses were carried out. The results indicate that the expression of a limited, distinctive set of genes was altered in the two apoptosis-prone clones, not in the resistant clone. That clone showed altered expression of different sets of genes, suggesting that a molecular switch converted patterns of gene expression between the two phenotypes: apoptosis-prone and apoptosis-resistant. The results are consistent with the hypothesis that altered expression of a distinctive network of genes after glucocorticoid administration ultimately triggers apoptosis of leukemic lymphoid cells. The altered genes identified provide new foci for study of their role in cell death. PMID- 12782124 TI - A new imprinted cluster on the human chromosome 7q21-q31, identified by human mouse monochromosomal hybrids. AB - We have previously established a series of human monochromosomal hybrids containing a single human chromosome of defined parental origin as an in vitro resource for the investigation of human imprinted loci. Using the hybrids with a paternal or maternal human chromosome 7, we determined the allelic expression profiles of 76 ESTs mapped to the human chromosome 7q21-q31. Seven genes/transcripts, including PEG10 which has previously been reported to be imprinted, showed parent-of-origin-specific expression in monochromosomal hybrids. One of the 6 candidate genes/transcripts, i.e., DLX5 was confirmed to be imprinted in normal human lymphoblasts and brain tissues by a polymorphic analysis. Thus, an imprinted domain has been newly defined in the region of human chromosome 7q21-q31 using human-mouse monochromosomal hybrids. PMID- 12782125 TI - Characterization of the OFD1/Ofd1 genes on the human and mouse sex chromosomes and exclusion of Ofd1 for the Xpl mouse mutant. AB - Oral-facial-digital type 1 (OFD1) syndrome is an X-linked dominant condition characterized by malformations of the face, oral cavity, and digits. The responsible gene, OFD1, maps to human Xp22 and has an unknown function. We isolated and characterized the mouse Ofd1 gene and showed that it is subject to X inactivation, in contrast to the human gene. Furthermore, we excluded a role for Ofd1 in the pathogenesis of the spontaneous mouse mutant Xpl, which had been proposed as a mouse model for this condition. Comparative sequence analysis demonstrated that OFD1 is conserved among vertebrates and absent in invertebrates. This analysis allowed the identification of evolutionarily conserved domains in the protein. Finally, we report the identification of 18 apparently nonfunctional OFD1 copies, organized in repeat units on the human Y chromosome. These degenerate OFD1-Y genes probably derived from the ancestral Y homologue of the X-linked gene. The high level of sequence identity among the different units suggests that duplication events have recently occurred during evolution. PMID- 12782126 TI - Genomic organization and expression of mouse Tpt1 gene. AB - The translationally controlled tumor protein (TCTP), also known as histamine releasing factor (HRF), is encoded by a gene (Tpt1) that is highly conserved throughout phylogeny. TCTP is implicated in cell growth, acute allergic response, and apoptosis. In the present study, seven putative Tpt1 genes with different chromosomal localizations were identified in the mouse genome. In six of them, analysis of the 5' and 3' untranslated regions revealed the presence of flanking direct repeats and residual poly(A) tails typical of pseudogenes. Only three of the seven genes can produce a protein of the expected molecular weight. We isolated the genomic DNA of these three genes to analyze their sequence, genomic organization, and in vitro promoter activity. We found that mouse Tpt1 is localized on chromosome 14 with a canonical intron-exon organization, a functional promoter, and only one transcript that is ubiquitously expressed in all tissues. PMID- 12782127 TI - Identification of a novel protein interacting with laforin, the EPM2a progressive myoclonus epilepsy gene product. AB - We have identified an interacting partner protein (encoded by the human EPM2AIP1 gene (approved symbol)) for laforin, the product of the EPM2A gene, which is mutated in an autosomal recessive form of adolescent progressive myoclonus epilepsy. The EPM2AIP1 gene was identified in a screen for laforin-interacting proteins with a human brain cDNA library using the yeast two-hybrid system. The specificity of the interaction was confirmed by coimmunoprecipitation of in vivo transfected protein and by using EPM2A deletion constructs. Subcellular colocalization of laforin and EPM2AIP1 protein was also demonstrated. The human EPM2AIP1 gene, corresponding to the KIAA0766 cDNA clone in the databases, was characterized and shown, like EPM2A, to be ubiquitously expressed. The gene, which comprises one large exon 1824 nucleotides in length and has alternative 3' untranslated regions, maps to human chromosome 3p22.1. The function is currently not known and extensive analyses do not reveal any homology to other proteins or any obvious structural motifs. Because genetic heterogeneity in Lafora disease has been described, mutational analysis of the EPM2AIP1 gene was performed on non EPM2A patients, but no mutations were found. The identification of this first binding partner for laforin promises to be an important step toward unraveling the underlying pathogenesis of this severest form of teenage-onset epilepsy. PMID- 12782128 TI - Identification of RELMgamma, a novel resistin-like molecule with a distinct expression pattern. AB - We have identified RELMgamma, a novel member of the resistin-like molecule/found in inflammatory zone (RELM/FIZZ) family in mice and rats. Microarray and real time RT-PCR experiments revealed a repression of RELMgamma mRNA in nasal respiratory epithelium of cigarette smoke-exposed versus untreated rats. The analysis of the physiological tissue-specific expression revealed highest expression in hematopoietic tissues, suggesting a cytokine-like role for RELMgamma. RELMgamma is most closely related to RELMalpha/FIZZ1. Despite the high similarity, the expression properties of the two genes are clearly distinct. While RELMgamma (approved symbol retnlg) is expressed in rat white adipose tissue, minute to no expression of RELMalpha was detected in that system. Thus, previous reports analyzing RELMalpha expression in rat adipose tissue might have been influenced by cross-hybridization with RELMgamma. Finally we could demonstrate that white adipose tissue of mice shows strong RELMalpha expression but only low levels of RELMgamma, indicating a species-specific gene regulation. PMID- 12782129 TI - Identification of two novel polycystic kidney disease-1-like genes in human and mouse genomes. AB - Mutations to the prototypical members of the two general classes of polycystins, polycystin-1 encoded by PKD1 and polycystin-2 encoded by PKD2, underlie autosomal dominant polycystic kidney disease. Here we report the identification of a pair of genes homologous to PKD1 from both the human and mouse genomes. PKD1L2 and PKD1L3 are located on human chromosome 16q22-q23 and mouse chromosome 8 and are alternatively spliced. The human and mouse forms of PKD1L2 are highly conserved, with each one consisting of 43 exons and approximately 2,460 codons. PKD1L3 shows regional sequence divergence, with the mouse form having two additional exons and a much larger exon 5. The predicted protein products of PKD1L2 and PKD1L3 contain the combination of GPS and PLAT/LH2 domains that uniquely define them as polycystin-1 family members. They are predicted to have 11 membrane-spanning regions with a large extracellular domain consistent with the proposed receptor function of this protein family. PKD1L2 and PKD1L3 contain strong ion channel signature motifs that suggest their possible function as components of cation channel pores. Polycystin-1-related proteins may not only regulate channels, but may actually be part of the pore-forming unit. PMID- 12782130 TI - Identification of eight genes encoding chemokine-like factor superfamily members 1-8 (CKLFSF1-8) by in silico cloning and experimental validation. AB - TM4SF11 is only 102 kb from the chemokine gene cluster composed of SCYA22, SCYD1, and SCYA17 on chromosome 16q13. CKLF maps on chromosome 16q22. CKLFs have some characteristics associated with the CCL22/MDC, CX3CL1/fractalkine, CCL17/TARC, and TM4SF proteins. Bioinformatics based on CKLF2 cDNA and protein sequences in combination with experimental validation identified eight novel genes designated chemokine-like factor superfamily members 1-8 (CKLFSF1-8). CKLFSF1-8 form gene clusters; the sequence identities between CKLF2 and CKLFSF1-8 are from 12.5 to 39.7%. Most of the CKLFSFs have alternative RNA splicing forms. CKLFSF1 has a CC motif and higher sequence similarity with chemokines than with any of the other CKLFSFs. CKLFSF8 shares 39.3% amino acid identity with TM4SF11. CKLFSF1 links the CKLFSF family with chemokines, and CKLFSF8 links it with TM4SF. The characteristics of CKLFSF2-7 are intermediate between CKLFSF1 and CKLFSF8. This indicates that CKLFSF represents a novel gene family between the SCY and the TM4SF gene families. PMID- 12782131 TI - The human DDX and DHX gene families of putative RNA helicases. AB - Nucleic acid helicases are characterized by the presence of the helicase domain containing eight motifs. The sequence of the helicase domain is used to classify helicases into families. To identify members of the DEAD and DEAH families of human RNA helicases, we used the helicase domain sequences to search the nonredundant peptide sequence database. We report the identification of 36 and 14 members of the DEAD and DEAH families of putative RNA helicases, including several novel genes. The gene symbol DDX had been used previously for both DEAD- and DEAH-box families. We have now adopted DDX and DHX symbols to denote DEAD- and DEAH-box families, respectively. Members of human DDX and DHX families of putative RNA helicases play roles in differentiation and carcinogenesis. PMID- 12782132 TI - Overlap of promoter and coding sequences in the mouse STOP gene (Mtap6). AB - The microtubule-associated proteins STOP are essential for synaptic plasticity and integrated brain function. The STOP gene (Mtap6) is composed of 4 exons and presents different developmental and tissue specific alternative transcripts resulting in the neuronal isoforms E- and N-STOP, and in the fibroblastic F-STOP isoform. We now characterize the transcription initiation sites for neuronal and non neuronal STOP transcripts. Our results show that there is a single neuronal specific promoter for transcription of E- and N-STOP mRNAs. F-STOP mRNA transcription is regulated by a distinct promoter. A remarkable feature of this promoter is that it lies within coding sequences expressed in neuronal E- and N STOP isoforms. PMID- 12782133 TI - A comparative genomic analysis of the cow, pig, and human CFTR genes identifies potential intronic regulatory elements. AB - The identification of sequences within noncoding regions of genes that are conserved between several species may indicate potential regulatory elements. This is important for genes with complex control mechanisms such as the cystic fibrosis transmembrane conductance regulator (CFTR). CFTR demonstrates similar patterns of temporal and spatial expression in human and sheep, but these differ significantly in mouse cftr. The complete sheep CFTR sequence is unavailable so we annotated BAC clones encompassing the CFTR gene from two other artiodactyl species (cow and pig) for comparative sequence analysis. Regions of introns 2, 3, 10, 17a, 18, and 21 and 3' flanking sequence corresponding to human CFTR DNase I hypersensitive sites (DHS) showed high homology in the cow and pig. Cross-species sequence conservation also enabled finer mapping of other human DHS, including those in introns 1, 16, and 20. Additional potential regulatory elements not associated with human DHS were also identified. PMID- 12782134 TI - Organization of the human FK506-binding immunophilin FKBP52 protein gene (FKBP4). AB - FKBP52 is a widely expressed FK506-binding immunophilin that possesses peptidylprolyl isomerase activity and a tetratricopeptide repeat involved in protein-protein interaction. FKBP52 plays an important role in steroid receptor function and is implicated in other diverse processes, including regulation of transcription, cation channel activity, and gene transfer efficiency. Reported here is the genomic organization of the human FKBP52 gene (FKBP4), which shares all but one of the same exon-intron boundaries as the structurally related immunophilin FKBP51 gene (FKBP5). Approximately 3.5 kb of 5'-flanking DNA of FKBP4 was subcloned into a luciferase reporter vector and was found to exhibit robust activity in T-47D, MCF7, and COS-7 cells. Promoter constructs with only 143 bp of upstream sequence maintained high activity. This region contains a CAAT motif sequence and consensus binding sites for Sp1, heat-shock factor, and MYC MAX, which are conserved in the rabbit FKBP4 promoter and, when deleted, dramatically reduced promoter activity in T-47D cells. PMID- 12782135 TI - Atp10a, the mouse ortholog of the human imprinted ATP10A gene, escapes genomic imprinting. AB - The mouse Atp10a gene is located at the border of an imprinted domain distal to the p-locus on mouse chromosome 7. The localization of Atp10a neighboring the maternally expressed gene Ube3a in the imprinted domain and an unusual inheritance pattern of the obesity phenotype with a p-locus deletion have suggested that Atp10a might be imprinted and associated with body fat. Recently, its human ortholog, ATP10A, was identified as the second imprinted gene with maternal expression in the human chromosome 15q11-q13 imprinted domain. To elucidate the imprinting status of Atp10a, we performed expression analysis in various tissues from reciprocal crosses between C57BL/6 and PWK (divergent strains of Mus musculus) mice. The results revealed that Atp10a was biallelically expressed in all tissues examined. Furthermore, there was no differential methylation in the CpG island and no antisense transcripts of the gene. These findings suggest that the mouse Atp10a gene escapes genomic imprinting. PMID- 12782137 TI - Cell signaling events: a view from the matrix. AB - Cellular activities are primarily initiated, modulated and sustained by multifunctional molecules (cytokines and growth factors) that are secreted into the extracellular space and that signal through membrane-bound, high-affinity receptors. In contrast to the fairly well understood mechanisms that mediate the specificity of signal transduction within the confined and compartmentalized environment of the cell, significantly less is known about the mechanisms that regulate the availability of signaling molecules in the extracellular milieu. Recent findings have implicated the participation of extracellular protein macroaggregates in signaling events controlling patterning and morphogenesis. The results suggest a functional coupling between the tissue-specific organization of collagenous and elastic macroaggregates and their ability to perform instructive as well as structural functions. These observations open the way to a novel understanding in these poorly understood and critically important areas of cell and developmental biology. PMID- 12782138 TI - Codistribution analysis of elastin and related fibrillar proteins in early vertebrate development. AB - Elastin is an extracellular matrix protein found in adult and neonatal vasculature, lung, skin and connective tissue. It is secreted as tropoelastin, a soluble protein that is cross-linked in the tissue space to form an insoluble elastin matrix. Cross-linked elastin can be found in association with several microfibril-associated proteins including fibrillin-1, fibrillin-2 and fibulin-1 suggesting that these proteins contribute to elastic fiber assembly, structure or function. To date, the earliest reported elastin expression was in the conotruncal region of the developing avian heart at 3.5 days of gestation. Here we report that elastin expression begins at significantly earlier developmental stages. Using a novel immunolabeling method, the deposition of elastin, fibrillin 1 and -2 and fibulin-1 was analyzed in avian embryos at several time points during the first 2 days of development. Elastin was found at the midline associated with axial structures such as the notochord and somites at 23 h of development. Fibrillin-1 and -2 and fibulin-1 were also expressed at the embryonic midline at this stage with fibrillin-1 and fibulin-1 showing a high degree of colocalization with elastin in fibers surrounding midline structures. The expression of these genes was confirmed by conventional immunoblotting and mRNA detection methods. Our results demonstrate that elastin polypeptide deposition occurs much earlier than was previously appreciated. Furthermore, the results suggest that elastin deposition at the early embryonic midline is accompanied by the deposition and organization of a number of extracellular matrix polypeptides. These filamentous extracellular matrix structures may act to transduce or otherwise stabilize dynamic forces generated during embryogenesis. PMID- 12782139 TI - Matrix deposition of tryptophan-containing allelic variants of type IX collagen in developing human cartilage. AB - Genetic polymorphisms that encode a tryptophan (Trp) residue in the triple helical domain of the alpha2 (Trp2) or alpha3 chain (Trp3) of human type IX collagen have been linked to risk of degenerative intervertebral disc disease. To determine whether these two allelic variants express protein that may affect the extracellular matrix of cartilage in vivo, we examined the properties of resident type IX collagen in an anonymous collection of embryonic and fetal human cartilage samples screened for Trp genotypes. No difference was found in the yield and electrophoretic properties of pepsin-solubilized type IX collagen between Trp2, Trp3 and non-Trp cartilage samples. On Western blot analysis, a polyclonal antiserum raised against a synthetic peptide matching the immediate Trp-containing sequence of the Trp3 allele reacted specifically with the alpha3(IX) chain prepared from Trp3 cartilage samples. Two-dimensional peptide mapping of type IX collagen in CNBr-digests of whole tissue gave indistinguishable fingerprints for Trp2, Trp3 and control tissues, including the yield of cross-linked peptides. Analysis of one cartilage sample that was homozygous for the Trp2 allele also gave a normal yield of collagen IX, including its alpha2 chain and a normal profile of cross-linked peptides. Together, the findings indicate that both Trp2 and Trp3 allelic products are incorporated into the cross-linked fibrillar network of developing human cartilage apparently normally. Any pathological consequences are likely, therefore, to be long-term and indirect rather than from overt misassembly of matrix. PMID- 12782140 TI - Discrete integration of collagen XVI into tissue-specific collagen fibrils or beaded microfibrils. AB - The structural and functional diversity of extracellular matrices is determined, not only by individual macromolecules, but even more decisively, by the alloyed aggregates they form. Although quantitatively major matrix molecules can occur ubiquitously, their organization varies from one tissue to another due to their amalgamation with specific sets of minor components. Here, we show that the fibril-associated collagen with interrupted triple helices collagen XVI is unique in that, depending on the tissue context, it can be incorporated into distinct suprastructural aggregates. In papillary dermis, the protein unexpectedly does not occur in banded collagen fibrils, but rather, is a component of specialized fibrillin-1-containing microfibrils. In territorial cartilage matrix, however, collagen XVI is not a component of aggregates containing fibrillin-1. Instead, the protein resides in a discrete population of thin, weakly banded collagen fibrils also containing collagens II and XI. Collagen IX also occurs in this population of fibrils, but at longitudinal locations discrete from those of collagen XVI. This suprastructural versatility of a collagen is without precedent and highlights pivotal differences in the tissue-specific organization of matrix aggregate structures. PMID- 12782141 TI - Crystal structure of the collagen alpha1(VIII) NC1 trimer. AB - Collagen VIII is a major component of Descemet's membrane and is also found in vascular subendothelial matrices. The C-terminal non-collagenous domain (NC1) domain of collagen VIII, which is a member of the C1q-like protein family, forms a stable trimer and is thought to direct the assembly of the collagen triple helix, as well as polygonal supramolecular structures. We have solved the crystal structure of the mouse alpha1(VIII)(3) NC1 domain trimer at 1.9 A resolution. Each subunit of the intimate NC1 trimer consists of a ten-stranded beta-sandwich. The surface of the collagen VIII NC1 trimer presents three strips of partially exposed aromatic residues shown to interact with the non-ionic detergent CHAPS, which are likely to be involved in supramolecular assemblies. Equivalent strips exist in the NC1 domain of the closely related collagen X, suggesting a conserved assembly mechanism. Surprisingly, the collagen VIII NC1 trimer lacks the buried calcium cluster of the collagen X NC1 trimer. The mouse alpha1(VIII) and alpha2(VIII) NC1 domains are 71.5% identical in sequence, with the differences being concentrated on the NC1 trimer surface. A few non-conservative substitutions map to the subunit interfaces near the surface, but it is not obvious from the structure to what extent they determine the preferred assembly of collagen VIII alpha1 and alpha2 chains into homotrimers. PMID- 12782142 TI - Changing profiles of proteoglycans in the transition of predentine to dentine. AB - Proteoglycans and their constituent glycosaminoglycans have been proposed to play important roles in matrix mediated formation of mineralised tissues, such as dentine. This study has examined the changing profile of proteoglycan species during the transition of unmineralised predentine to mineralised dentine. Three week-old calves teeth were collected and proteoglycans purified from the predentine, the predentine/dentine interface and dentine. Decorin and biglycan, together with related degradation products, were identified in the predentine fraction, alongside degradation products of versican, indicating metabolism of the proteoglycan components within this tissue. Decorin and biglycan were also identified as major proteoglycan species within extracts from the predentine/dentine interface and dentine. Analysis of the glycosaminoglycan constituents within each fraction demonstrated significant changes in their composition. Predentine contained a high proportion of dermatan sulfate (DS) (51.5%), with chondroitin sulfate (CS) (17.8%) and hyaluronan (HA) (30.7%) additionally identified. Within the predentine/dentine interface the proportion of CS increased greatly (62.5%), with corresponding decrease in the proportion of DS (21.4%) and HA (16.1%) also evident. CS only was identifiable within the dentine matrix. A four-fold increase in the level of sulfation was identified for glycosaminoglycans extracted from the predentine/dentine interface compared with the predentine and dentine fraction. The ratio of DeltaDi4S:DeltaDi6S was higher for glycosaminoglycans isolated from the predentine fraction. Glycosaminoglycans extracted from the dentine fraction possessed longer chain lengths than those present in the predentine and predentine/dentine fractions. The results indicate that the proteoglycans within each fraction undergo subtle structural modification, particularly at the onset of mineralisation, indicating an active involvement of these macromolecules in the overall mineralisation process. PMID- 12782143 TI - Syndecan-1 accumulates in lysosomes of poorly differentiated breast carcinoma cells. AB - Expression patterns of syndecan-1, the cell surface heparan sulfate proteoglycan (HSPG) predominant on epithelial cells, were analyzed in tissue samples from 30 infiltrating human breast carcinomas and in 9 human breast carcinoma cell lines. Immunohistochemical staining demonstrates that while a subset of the breast carcinomas lose syndecan-1, this proteoglycan is expressed or overexpressed in a majority of the cases. Interestingly, cells in poor grade tumors contain intracellular syndecan-1, an observation that has not been previously described and was thus further investigated. Examination of cultured breast carcinoma cell lines indicates that they also display the phenotype of the syndecan-1 positive tumors and thereby provide a model system for analysis of intracellular syndecan 1. All cell lines examined express syndecan-1, and poorly differentiated lines such as BT549 cells internalize the proteoglycan from the cell surface where it accumulates as intact HSPG in intracellular vesicles. Colocalization studies using fluorescent markers identify these to be lysosomes. This finding is unexpected, as the accepted mechanism for degradation of syndecan HSPG following endocytosis is fragmentation of the protein core and glycosaminoglycan chains in endosomes, followed by delivery of the fragments to lysosomes. Lysosomal inactivation using ammonium chloride demonstrates that well-differentiated lines such as T47D and MCF-7 cells, which maintain the majority of syndecan-1 on their cell surfaces, also target intact constitutively endocytosed syndecan-1 to lysosomes. Taken together, these results suggest that mammary epithelial cells utilize a previously uncharacterized mechanism for syndecan-1 catabolism. In this pathway the proteoglycan remains intact as it passes through the endosomal system, prior to arriving at its site of intracellular degradation in lysosomes. PMID- 12782144 TI - Calreticulin--an endoplasmic reticulum protein with calcium-binding activity is also found in the extracellular matrix. AB - Previous studies have reported that calreticulin (CRT), a calcium-binding and chaperoning protein, is expressed only in the endoplasmatic reticulum, nucleus and at the cell surface. In this study we clearly show that odontoblasts and predentin matrix contain CRT. To our knowledge, this is the first time CRT has been described in the extracellular matrix. The expression of CRT was studied by immunohistochemistry, ultrastructural immunocytochemistry and in situ hybridization in developing rat teeth. CRT was detected as a 59-kDa protein in rat pulp cell culture medium and dentin extracellular matrix extract by Western blotting. The presence of the protein was shown in rat odontoblasts and predentin with immunohistochemistry. At the ultrastructural level, the labeling was distributed in the rat odontoblasts, ameloblasts and predentin. Northern blotting showed the presence of CRT mRNA in rat molars, which was confirmed by in situ hybridization in odontoblasts and ameloblasts. We now present the first convincing evidence that CRT is found in extracellular matrix where it may play an important role in mineralization. PMID- 12782145 TI - Paxillin-associated focal adhesion involvement in perinatal pulmonary arterial remodelling. AB - Birth is followed by remodelling of the actin cytoskeleton of pulmonary arterial smooth muscle cells, then by extracellular matrix deposition. Hypothesising that the cell/matrix adhesions would also be remodelled, we investigated the expression, localisation and biochemical characteristics of the focal adhesion protein paxillin in vivo, in vessels from normal and pulmonary hypertensive neonatal piglets. Initially we showed that in intact porcine pulmonary arteries exposed to cytochalasin D there was a reduction filamentous actin accompanied by a reduction in paxillin-associated focal adhesions, similar to that seen in cultured pulmonary arterial smooth muscle cells. Vessels from normal and hypoxic animals were found to have two isoforms of paxillin, of 60 and 66 kDa with pI values of 6.7-4.2. Transient changes occurred during the first 14 days of life. Between birth and 6 days there was a reduction in the amount of both paxillin isoforms, a shift to more acidic pI values and an increase in paxillin phosphorylation. Simultaneously, immunostaining showed a transient reduction in paxillin expression, a change temporally and spatially associated with a previously demonstrated reduction in actin. Findings are consistent with an immediate postnatal spatial reorganisation of paxillin-associated focal adhesions. Paxillin content and remodelling was abnormal in pulmonary hypertensive arteries, the response varying according to postnatal age. PMID- 12782146 TI - Plant sphingolipids: structural diversity, biosynthesis, first genes and functions. AB - In mammals and Saccharomyces cerevisiae, sphingolipids have been a subject of intensive research triggered by the interest in their structural diversity and in mammalian pathophysiology as well as in the availability of yeast mutants and suppressor strains. More recently, sphingolipids have attracted additional interest, because they are emerging as an important class of messenger molecules linked to many different cellular functions. In plants, sphingolipids show structural features differing from those found in animals and fungi, and much less is known about their biosynthesis and function. This review focuses on the sphingolipid modifications found in plants and on recent advances in the functional characterization of genes gaining new insight into plant sphingolipid biosynthesis. Recent studies indicate that plant sphingolipids may be also involved in signal transduction, membrane stability, host-pathogen interactions and stress responses. PMID- 12782147 TI - Serine palmitoyltransferase, a key enzyme of sphingolipid metabolism. AB - The first step in the biosynthesis of sphingolipids is the condensation of serine and palmitoyl CoA, a reaction catalyzed by serine palmitoyltransferase (SPT) to produce 3-ketodihydrosphingosine (KDS). This review focuses on recent advances in the biochemistry and molecular biology of SPT. SPT belongs to a family of pyridoxal 5'-phosphate (PLP)-dependent alpha-oxoamine synthases (POAS). Mammalian SPT is a heterodimer of 53-kDa LCB1 and 63-kDa LCB2 subunits, both of which are bound to the endoplasmic reticulum (ER) most likely with the type I topology, whereas other members of the POAS family are soluble homodimer enzymes. LCB2 appears to be unstable unless it is associated with LCB1. Potent inhibitors of SPT structurally resemble an intermediate in a probable multistep reaction mechanism for SPT. Although SPT is a housekeeping enzyme, its activity is regulated transcriptionally and post-transcriptionally, and its up-regulation is suggested to play a role in apoptosis induced by certain types of stress. Specific missense mutations in the human LCB1 gene cause hereditary sensory neuropathy type I, an autosomal dominantly inherited disease, and these mutations confer dominant-negative effects on SPT activity. PMID- 12782148 TI - Effects of plasma apolipoproteins on lipoprotein lipase-mediated lipolysis of small and large lipid emulsions. AB - Large (ca. 120 nm) and small (ca. 35 nm) emulsions consisting of triolein (TO) and phosphatidylcholine (PC) were prepared as the primary protein-free models of chylomicrons and their remnants, respectively. Lipoprotein lipase (LPL)-mediated lipolysis of emulsion TO was retarded in chylomicron-free human plasma compared with the hydrolysis activated by isolated apolipoprotein C-II (apoC-II). In 30% plasma, free fatty acid (FFA) release rate was higher for large emulsions than for small ones, while both emulsions were hydrolyzed at similar rates in the presence of isolated apoC-II. Isolated apolipoprotein C-III (apoC-III) or apolipoprotein E (apoE) worked as LPL-inhibitor of the lipolysis activated by apoC-II. It was also observed that apolipoprotein A-I (apoA-I) showed distinct inhibitory effects on the lipolysis of large and small emulsions: more effective inhibition for small emulsions. Kinetic analyses showed that K(m)(app) and V(max)(app) for the lipolysis of emulsions were lower in the presence of 30% plasma than isolated apoC-II. ApoA-I also markedly decreased K(m)(app) and V(max)(app) for LPL-catalyzed hydrolysis of both emulsions. In chylomicron-free serum, the density of bound apoA-I at small emulsion surfaces was about three fold greater than large emulsion surfaces, but the binding densities of apoC-II, apoC-III and apoE were less for small emulsion surfaces than for large ones, suggesting that apoA-I preferentially binds to small particles and displaces other exchangeable apolipoproteins from particle surfaces. These results indicate that, in addition to the well known inhibitory effects of apoC-III and apoE, apoA I in plasma regulates the lipolysis of triglyceride (TG)-rich emulsions and lipoproteins in a size-dependent manner. PMID- 12782149 TI - Metabolism of 25-hydroxyvitamin D3 by microsomal and mitochondrial vitamin D3 25 hydroxylases (CYP2D25 and CYP27A1): a novel reaction by CYP27A1. AB - The metabolism of 25-hydroxyvitamin D(3) was studied with a crude mitochondrial cytochrome P450 extract from pig kidney and with recombinant human CYP27A1 (mitochondrial vitamin D(3) 25-hydroxylase) and porcine CYP2D25 (microsomal vitamin D(3) 25-hydroxylase). The kidney mitochondrial cytochrome P450 catalyzed the formation of 1alpha,25-dihydroxyvitamin D(3), 24,25-dihydroxyvitamin D(3) and 25,27-dihydroxyvitamin D(3). An additional metabolite that was separated from the other hydroxylated products on HPLC was also formed. The formation of this 25 hydroxyvitamin D(3) metabolite was dependent on NADPH and the mitochondrial electron transferring protein components. A monoclonal antibody directed against purified pig liver CYP27A1 immunoprecipitated the 1alpha- and 27-hydroxylase activities towards 25-hydroxyvitamin D(3) as well as the formation of the unknown metabolite. These results together with substrate inhibition experiments indicate that CYP27A1 is responsible for the formation of the unknown 25-hydroxyvitamin D(3) metabolite in kidney. Recombinant human CYP27A1 was found to convert 25 hydroxyvitamin D(3) into 1alpha,25-dihydroxyvitamin D(3), 25,27-dihydroxyvitamin D(3) and a major metabolite with the same retention time on HPLC as that formed by kidney mitochondrial cytochrome P450. Gas chromatography-mass spectrometry (GC MS) analysis of the unknown enzymatic product revealed it to be a triol different from other known hydroxylated 25-hydroxyvitamin D(3) metabolites such as 1alpha,25-, 23,25-, 24,25-, 25,26- or 25,27-dihydroxyvitamin D(3). The product had the mass spectrometic properties expected for 4beta,25-dihydroxyvitamin D(3). Recombinant porcine CYP2D25 converted 25-hydroxyvitamin D(3) into 1alpha,25 dihydroxyvitamin D(3) and 25,26-dihydroxyvitamin D(3). It can be concluded that both CYP27A1 and CYP2D25 are able to carry out multiple hydroxylations of 25 hydroxyvitamin D(3). PMID- 12782150 TI - Inhibition of cholesterol crystallization under bilirubin deconjugation: partial characterization of mechanisms whereby infected bile accelerates pigment stone formation. AB - Pigment gallstones have been reported to be closely associated with biliary tract infection. We previously reported that addition of unconjugated bilirubin (UCB), which is deconjugated by beta-glucuronidase in infected bile, could enhance cholesterol crystal formation in supersaturated model bile (MB). The present study evaluated the effect of beta-glucuronidase on the processes of pigment gallstone formation and cholesterol crystallization. Supersaturated MB (taurocholate/lecithin/cholesterol at 71:18:11, a total lipid concentration of 10.0 g/dl and a cholesterol saturation index (CSI) of 2.0) and native rat bile were mixed at a ratio of 3:1. Then, mixed bile was incubated with or without beta glucuronidase and changes of the following parameters were investigated over time: (1) the UCB/total bilirubin ratio; (2) cholesterol crystal formation; (3) the precipitate weight and the cholesterol concentration in the precipitate and supernatant; and (4) the lipid distribution of vesicles in the supernatant. Compared with beta-glucuronidase-free bile, (1) beta-glucuronidase-containing bile showed a significant increase of the UCB/total bilirubin ratio, (2) as well as a significantly longer nucleation time (96+/-17.0 vs. 114+/-20.0) and fewer cholesterol crystals. (3) The precipitate weight and the cholesterol concentration in the precipitate were significantly increased, while the cholesterol concentration in supernatant was decreased. (4) When mixed bile was incubated with beta-glucuronidase, the cholesterol concentration in the vesicles was lower than in bile without beta-glucuronidase. The precipitate weight and the cholesterol concentration in the precipitate was increased by incubation with beta-glucuronidase, while cholesterol concentration was decreased in the supernatant (especially in the vesicles). This means that bile vesicles were more stable and it was more difficult for cholesterol crystals to form. Thus, the presence of beta-glucuronidase may inhibit the formation of pure cholesterol stones even in the presence of cholesterol supersaturation. PMID- 12782151 TI - Long-chain fatty acyl-CoA esters induce lipase activation in the absence of a water-lipid interface. AB - In most lipases a mobile element or lid domain covers the catalytic site of the enzyme and the lid opening event, which usually proceed at a lipid-water interface, is required to form the catalytically competent lipase. We report here a noticeable increase in activity of two fungal lipases assayed in aqueous solution in absence of any interface when adding submicellar concentrations of amphipathic physiological molecules like long-chain acyl-CoAs. The catalytic activity was dramatically dependent on the acyl chain length of the amphiphile and could be related with a lid-opening process. Our data support that lipase activation can be triggered in the absence of a well-defined interface, and stresses the notion that other non-aggregated amphipathic constituents of the local microenvironment can act as putative regulators of lipase activity. PMID- 12782152 TI - Participation of phospholipase D and alpha/beta-protein kinase C in growth factor induced signalling in C3H10T1/2 fibroblasts. AB - We have studied phospholipase D (PLD) activation in relation to protein kinase C (PKC) and the involvement of PLD in extracellularly regulated kinase 1 (MAPK) (ERK1) activation and c-fos mRNA expression in C3H/10T1/2 (Cl8) fibroblasts. In these cells, the PLD activity was significantly increased by porcine platelet derived growth factor (PDGF-BB), phorbol 12-myristate 13-acetate (PMA), and epidermal growth factor (EGF). PLD activation by PDGF-BB and PMA, but not EGF, was inhibited in Cl8 cells expressing the HAbetaC2-1 peptide (Cl8 HAbetaC2-1 cells), with a sequence (betaC2-1) shown to bind receptor for activated C kinase 1 (RACK1) and inhibit c-PKC-mediated cell functions [Science 268 (1995) 247]. A role of alpha-PKC in PLD activation is further underscored by co immunoprecipitation of alpha-PKC with PLD1 and PLD2 in non-stimulated as well as PMA- and PDGF-BB-stimulated Cl8 cells. However, only PKC in PLD1 precipitates was activated by these agonists, while the PKC in the PLD2 precipitates was constitutively activated. The c-fos mRNA levels in Cl8 cells increased more than 30-fold in response to either PDGF-BB, EGF, or PMA. Approximately 60% inhibition of this increase in c-fos mRNA levels was observed in Cl8 HAbetaC2-1 cells. Formation of phosphatidylbutanol (PtdBut) at the expense of phosphatidic acid (PtdH) in the presence of n-butanol inhibited ERK1 activation and c-fos mRNA expression in PDGF-BB-treated Cl8 cells. ERK activation by PMA was unaffected by n-butanol in Cl8 cells but almost abolished by n-butanol in Cl8 HAbetaC2-1 cells, showing that ERK activation by PMA is heavily dependent on PKC and PLD1. In contrast, ERK activation by EGF in both cell types was not sensitive to n butanol. These results indicate (1) a role of a functional interaction between the RACK1 scaffolding protein and a alphaPKC-PLD complex for achieving full PLD activity in PDGF-BB- and PMA-stimulated Cl8 cells; (2) PLD-mediated PtdH formation is needed for optimal ERK1 activation by PDGF-BB and maximal increase in c-fos mRNA expression. These findings place PLD as an important component in PDGF-BB- and PMA-stimulated intracellular signalling leading to gene activation in Cl8 cells, while EGF does not require PLD. PMID- 12782153 TI - Gene expression and molecular composition of phospholipids in rat brain in relation to dietary n-6 to n-3 fatty acid ratio. AB - Rats were fed from conception till adulthood either with normal rat chow with a linoleic (LA) to linolenic acid (LNA) ratio of 8.2:1 or a rat chow supplemented with a mixture of perilla and soy bean oil giving a ratio of LA to LNA of 4.7:1. Fat content of the feed was 5%. Fatty acid and molecular species composition of ethanolamine phosphoglyceride was determined. Effect of this diet on gene expression was also studied. There was an accumulation of docosahexaenoic (DHA) and arachidonic acids (AA) in brains of the experimental animals. Changes in the ratio sn-1 saturated, sn-2 docosahexaenoic to sn-1 monounsaturated, sn-2 docosahexaenoic were observed. Twenty genes were found overexpressed in response to the 4.7:1 mixture diet and four were found down-regulated compared to normal rat chow. Among them were the genes related to energy household, lipid metabolism and respiration. The degree of up-regulation exceeded that observed with perilla with a ratio of LA to LNA 8.2:1 [Proc. Natl. Acad. Sci. U. S. A. 99 (2002) 2619]. It was concluded that brain sensitively reacts to the fatty acid composition of the diet. It was suggested that alteration in membrane architecture and function coupled with alterations in gene expression profiles may contribute to the observed beneficial impact of n-3 type polyunsaturated fatty acids on cognitive functions. PMID- 12782154 TI - Bezafibrate is a dual ligand for PPARalpha and PPARbeta: studies using null mice. AB - Bezafibrate is a known activator of peroxisome proliferator-activated receptors (PPARs) that can activate both PPARalpha and PPARbeta. To determine the role(s) of these receptors in mediating the biological effects of this chemical, the effect of bezafibrate was examined in PPARalpha-null and PPARbeta-null mice. Wild type, PPARalpha-null, or PPARbeta-null mice were fed either a control diet or one containing 0.5% bezafibrate for 10 days. Bezafibrate feeding caused a significant increase in liver weight in wild-type and PPARbeta-null mice compared to controls, while liver weight was unchanged in bezafibrate-fed PPARalpha-null mice. Gonadal adipose stores were significantly smaller in wild-type and PPARbeta null mice fed bezafibrate than in controls, and this effect was not found in similarly fed PPARalpha-null mice. Analysis of liver, white adipose tissue, and intestinal mRNAs showed that bezafibrate caused similar changes of mRNAs encoding lipid metabolizing enzymes in wild-type and PPARbeta-null mice compared to controls. Interestingly, in PPARalpha-null mice, bezafibrate also induced several mRNAs previously thought to be solely controlled by PPARalpha, showing that the effects of this drug are not exclusively modulated by this PPAR isoform. Western blot analysis of liver protein was consistent with changes in mRNA expression showing that the alterations in mRNA expression correlate with protein expression in this tissue. Results from these studies demonstrate that the effect of bezafibrate is mediated in large part by PPARalpha, although some changes in gene expression are dependent on PPARbeta. In contrast to other PPARalpha ligands such as WY-14,643, induction of some target genes by bezafibrate can also be modulated in the absence of a functional PPARalpha. PMID- 12782155 TI - Structure elucidation of phenolic compounds from red/white wine with antiatherogenic properties. AB - The oxidative modification of low-density lipoproteins (LDL) is supposed to play a critical role in atherogenesis. During this oxidation a potent inflammatory phospholipid mediator named platelet activating factor (PAF) is produced, and it is believed to be the key for the initiation of the inflammation and therefore for the process of atherogenesis. From many studies, it is established that wine has beneficial effects on health, including protection against cardiovascular diseases. According to our point of view, the cardioprotective effect of wine may be attributed partly to the existence of PAF antagonists in red or white wine and partly to the existence of antioxidants that reduce the oxidation of LDL and therefore the production of PAF. In this study, wine compounds that antagonize PAF were isolated and purified via chromatographic procedures, and determined structurally using chemical, enzymatic and spectroscopic methods. PMID- 12782156 TI - Morphological study of corneal endothelium and corneal thickness in pseudoexfoliation syndrome. AB - PURPOSE: To examine the endothelial morphology and the central thickness of the cornea in the eyes of patients with pseudoexfoliation syndrome (PXS). METHODS: The corneal endothelium was investigated in 21 patients (26 PXS eyes; 7 eyes of glaucoma patients, and 19 eyes of patients without glaucoma) and in 30 patients with senile cataract or refractive errors who served as the control group (30 non PXS eyes). The corneal endothelial morphology and central corneal thickness (CCT) were compared between the two groups. RESULTS: The corneal endothelial cell density was significantly lower in the PXS eyes (2336+/-383 cells/mm(2)) than in the non-PXS eyes (2632+/-327 cells/mm(2)) (P=.003). There was no significant difference in the coefficient of variation for the cell area and the percentage of hexagonal cells between these two groups. The central cornea was significantly thinner in the PXS eyes (529+/-31 micro m) than in the non-PXS eyes (547+/-28 micro m) (P=.03). There were no significant differences in these factors between the PXS eyes in patients with and without glaucoma. CONCLUSIONS: In PXS eyes, regardless of the presence of glaucoma in the patients, the corneal endothelial cell density is decreased and the central cornea is thin. PMID- 12782157 TI - The use of competitive PCR for quantitation of HSV-1 DNA. AB - PURPOSE: Polymerase chain reaction (PCR) detects the genomic materials of etiological agents with high specificity and sensitivity. However, in herpes simplex virus type 1 (HSV-1) infection, the clinical significance of the results often poses controversy because of the subclinical viral shedding during latent infection. Quantitative PCR might provide additional information to help clinical evaluation of the results. METHODS: Virus DNA was extracted from high titer stock of human HSV-1 (FK 25) by phenol/chloroform treatment. Construct (p/HSV-1) was made by inserting the glycoprotein D gene obtained from virus DNA into p(GEM-T) vector. Competitor (p/DeltaHSV-1) was made by deleting the inner 40 bp of construct (p/HSV-1) with restriction enzyme. Competitive PCR was performed using primers that amplify the glycoprotein D gene, and a template made of a 1:1 molar mixture of HSV-1 DNA and the competitor. RESULTS: The PCR product reflected the initial template dose from 20 to 30 cycles. Minimum detection level of HSV-1 DNA was 0.01 ng. CONCLUSION: Competitive PCR can quantitate HSV-1 DNA. PMID- 12782158 TI - A novel mutation of the TGFBI gene found in a Vietnamese family with atypical granular corneal dystrophy. AB - BACKGROUND: Mutation of the human transforming growth factor beta-induced (TGFBI) gene causes granular corneal dystrophy (GCD) in various ethnic groups. In this report, we identify the genetic defect on the TGFBI gene in a Vietnamese family with atypical GCD . CASES: The patient and her relatives were examined clinically. Genomic DNA was extracted from blood leukocytes. Fifty normal Vietnamese were used as controls. Analysis of the TGFBI gene was performed using polymerase chain reaction and direct sequencing. OBSERVATIONS: The 42-year-old proband clinically showed multiple white dot-like opacities scattered in the anterior and mid-stroma of the central cornea. Unlike GCD, these deposits were smaller, localized deeper and less severe. DNA analysis revealed a nucleotide transversion at codon 123 (GAC --> CAC), causing Asp --> His substitution (D123H). This mutation was also detected in 3 out of 5 unaffected family members, but was absent in the 50 normal controls. CONCLUSIONS: The novel D123H mutation of the TGFBI gene was not co-segregated with GCD in the family studied, and did not exist in the control population. It probably was a disease-causing mutation, thus expected to cause a novel variant of GCD in the proband. The detection of the D123H mutation in three unaffected family members indicates that it has low penetrance for GCD. PMID- 12782159 TI - Effects of topical instillation of traditional herbal medicines, herbal extracts, and their components on prostaglandin E2-induced aqueous flare elevation in pigmented rabbits. AB - PURPOSE: To evaluate the effect of topical instillation of traditional herbal medicines, herbal extracts, and their components on the elevation of aqueous flare induced by prostaglandin E(2) (PGE(2)) in pigmented rabbits. METHODS: Transcorneal diffusion of 25 micro g/mL of PGE(2) was carried out through a glass cylinder placed on the cornea to induce aqueous flare elevation in pigmented rabbits. Traditional herbal medicines, herbal extracts, and their components were topically instilled before the PGE(2) application. Aqueous flare was measured with a laser flare-cell meter. RESULTS: Two instillations, 60 and 30 minutes before PGE(2), of Kakkon-to, Sairei-to, Orengedoku-to, Senkanmeimoku-to, Scutellariae radix extract, Coptidis rhizoma extract, Gardeniae fructus extract, Phellodendri cortex extract, baicalein, baicalin, wogonin, crocetin, berberine, or glycyrrhizine did not inhibit the elevation induced by PGE(2). Two instillations, 60 and 30 minutes before PGE(2), of a Ligusticum wallichii extract (100 mg/mL) inhibited the elevation by 20%. Two instillations (5 and 3 hours before PGE(2)) of baicalein (1 mg/mL) or baicalin (5 mg/mL) inhibited the elevation by 16% and 24%, respectively. Two instillations, 5 and 3 hours before PGE(2), of wogonin, crocetin, berberine, or glycyrrhizine did not inhibit the elevation. CONCLUSION: Two instillations of Ligusticum wallichii extract 60 and 30 minutes before the PGE(2), and two instillations of baicalein or baicalin, 5 and 3 hours before the PGE(2), inhibited the PGE(2)-induced aqueous flare elevation in pigmented rabbits. PMID- 12782160 TI - Influence of cyclosporin on steroid-induced cataracts after renal transplantation. AB - PURPOSE: To study the effect of cyclosporin (CsA) on steroid-induced cataracts in patients following renal transplantation. METHODS: The subjects comprise 140 patients who had undergone renal transplantation at Kanazawa Medical Hospital. These subjects had received ophthalmologic examinations prior to their surgery and subsequently over the span of 12 months following surgery. The charts of these patients were re-examined for this study. The subjects were divided into two groups: the conventional therapy group (Group C) that received azathioprine and methylprednisolone; and the triple therapy group (Group T) that received azathioprine, methylprednisolone, and CsA. There were 73 and 67 patients in Groups C and T, respectively. The cataracts were classified according to Crews' classification. Steroid-induced cataract was diagnosed when vacuoles were observed or opacity was evident in the posterior subcapsular region. Subjects who exhibited any lens opacity before renal transplantation were excluded from this study. RESULTS: The total amounts of systemic steroid administered during the first year and at the final observation period were significantly lower in Group T than in Group C. The prevalence of steroid-induced cataracts of grade 1 and above was 55% and 63% for the first year, 73% and 89% for the second year, 74% and 92% for the third year, and 83% and 96% for the fifth year in Groups C and T, respectively. By the third year, the percentage of subjects exhibiting corrected visual acuity <0.8 or receiving cataract surgery was significantly higher in Group T than in Group C. There was no significant difference in the total dose of steroids, in subjects with cataracts over grade 1 and those with grades 0 and 1, between these groups. In Group C, the total steroid pulse therapy dose was significantly higher in subjects with steroid-induced cataracts over grade 1 than in those with grades 0 and 1. CONCLUSIONS: The observed rate of steroid-induced cataracts increased with the use of cyclosporin, despite a decrease in the total dose of systemic steroids. From this finding we suggest that the additional use of CSA may contribute to the development of steroid-induced cataracts. Steroid pulse therapy is considered a risk factor for the development of steroid-induced cataracts. PMID- 12782161 TI - Multiplex polymerase chain reaction for detection of herpes simplex virus type 1, type 2, cytomegalovirus, and varicella-zoster virus in ocular viral infections. AB - PURPOSE: To detect simultaneously herpes simplex virus type 1 (HSV-1), type 2 (HSV-2), cytomegalovirus (CMV), and varicella-zoster virus (VZV) in ocular specimens suspected of indicating viral infection, and to compare the results of multiplex polymerase chain reaction (PCR) with those of uniplex PCR. METHODS: Forty specimens, collected from 33 patients with clinically suspected herpes virus ocular infection, were tested. DNA was extracted from the specimens and amplified by multiplex and uniplex PCR. RESULTS: Both multiplex PCR and uniplex PCR gave the same results. Nineteen (19/33, 57.6%) patients were PCR-positive, among whom HSV-1 was detected in 13 (13/19, 68.4%) patients, and VZV in 6 (6/19, 31.6%) patients. CONCLUSION: These results demonstrated that multiplex PCR is as reliable as uniplex PCR, and is an accurate and a cost-saving method to identify several agents from a single specimen. PMID- 12782162 TI - Retinal S-antigen-reactive lymphocytes in a patient with uveitis associated with myelodysplastic syndromes. AB - BACKGROUND: Although autoimmune humoral abnormalities have been reported in patients with myelodysplastic syndromes (MDS), abnormal organ-specific cellular autoimmunity has not been demonstrated. METHODS: Peripheral blood lymphocytes (PBLs) and serum were collected from a uveitis patient with MDS. Cellular immune response against retinal S-antigen (S-Ag) was assessed by proliferation assay, and humoral immune response to S-Ag was measured by enzyme-linked immunosorbant assay. RESULTS: PBLs from the patient exhibited vigorous proliferation against S Ag, while humoral immune response against S-Ag was not detectable. PBLs from controls did not proliferate against S-Ag. CONCLUSION: These results provide new evidence that abnormal cellular immune responses against autoantigens may develop in MDS patients, thus leading to organ-specific autoimmune diseases such as uveitis, in addition to other systemic autoimmune disorders. PMID- 12782163 TI - Hepatocyte growth factor stimulates proliferation and migration during wound healing of retinal pigment epithelial cells in vitro. AB - PURPOSE: A defect in retinal pigment epithelial (RPE) cells may cause dysfunction of the neural retina, so rapid recovery of differentiated RPE cells is required after RPE injury. We investigated the effect of hepatocyte growth factor (HGF) on wound healing in RPE cells. METHODS: Confluent monolayers of bovine RPE cells were denuded, and the cells were allowed to recover in the presence or absence of HGF. The effect of HGF on RPE cell proliferation was evaluated by a 3-(4;5 dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulphophenyl)-2H-tetraz olium assay. In a migration assay, mitomycin C was used to inhibit proliferation, and the number of migrated cells was counted. The signaling pathways involved were examined using inhibitors of mitogen-activated protein kinase (MAPK), phosphatidylinositol-3 (PI3) kinase and protein kinase C pathways. RESULTS: At 80 ng/mL, HGF stimulated the wound closure of RPE monolayers and rendered the restituted cells more epithelioid in shape. HGF at 10 ng/mL stimulated RPE cell migration the most, whereas 80 ng/mL of HGF inhibited migration, but stimulated proliferation the most. In particular, PI3 kinase and MAPK inhibitor inhibited PRE cell migration and proliferation, respectively. CONCLUSIONS: HGF stimulated wound closure in cultured RPE cells, and rendered restituted cells epithelioid in shape. HGF may become a therapeutic candidate for RPE wound healing. PMID- 12782164 TI - Use of orbital color Doppler imaging for detecting internal carotid artery stenosis in patients with amaurosis fugax. AB - PURPOSE: To determine whether orbital color Doppler imaging (CDI) can be used to detect internal carotid artery stenosis (ICAS) in patients with amaurosis fugax (AF). METHODS: Twenty patients with AF (20 symptomatic eyes) and 14 normal controls (14 randomly selected eyes) were studied. The AF patients were divided by magnetic resonance angiography or carotid arteriography into those without ICAS (Group 1, n=14), and those with ICAS (Group 2, n=6). The peak systolic velocities (PSV) and end diastolic velocities (EDV) of the ophthalmic artery (OA) and the central retinal artery (CRA) were determined by orbital CDI, and the values were compared between the three groups. RESULTS: There were no significant differences in the PSV and EDV of the CRA between the three groups. However, the PSV and EDV of the OA were significantly lower in Group 2 than in the other two groups. From the distribution of PSV-OA and EDV-OA in AF patients, Group 2 could be separated from Group 1 by the PSV-OA value of 10 cm/s. CONCLUSIONS: The PSV-OA of 10 cm/s can be used to identify AF patients with ICAS. It is concluded that orbital CDI is a valuable method for detecting ICAS in AF patients. PMID- 12782165 TI - A muscle transposition procedure for abducens palsy, in which the halves of the vertical rectus muscle bellies are sutured onto the sclera. AB - PURPOSE: To review the results of a muscle transposition procedure in which the halves of the vertical rectus muscle bellies are sutured onto the sclera, without tenotomy of vertical recti as in Hummelsheim's procedure or surgical treatment of the lateral rectus (LR) as in Jensen's procedure. METHODS: Ten patients with abducens palsy received the procedure. We measured the ocular deviation and the field of single binocular vision, and observed the LR using magnetic resonance imaging (MRI). RESULTS: Preoperative or postoperative deviation was distributed from +27 to +58 prism diopters (PD) or orthophoria to +12 PD, respectively, in 7 patients with unilateral paresis, and +75 to +120 PD or +2 to +37 PD in 3 patients with bilateral paresis. The average correction was 42.4 PD per eye. Seven patients were able to regain the field of single binocular vision at least in the primary position. No postoperative complications were observed. MRI showed that the LR was atrophic and floppy, lacking muscle tension. CONCLUSIONS: Our procedure enabled the patients to obtain satisfactory postoperative results without treatment of the LR or tenotomy of the transposed muscles. This procedure can reduce operative damage to the eye more than Hummelsheim's or Jensen's procedure. PMID- 12782167 TI - Accommodative lag under habitual seeing conditions: comparison between adult myopes and emmetropes. AB - PURPOSE: To clarify whether myopes show poor accommodative response and thus have a larger accommodative lag under natural seeing conditions. METHODS: Forty-three adults without other ocular abnormalities were classified into the early-onset myopia (EOM, n=28) and the emmetropia (EMM, n=15) groups. The subjects wore glasses or contact lenses that they habitually used, and accommodative responses to four accommodative targets (16.0-50.5 cm from their eyes) were measured under a monocular or binocular condition using an open-field infrared autorefractometer. RESULTS: Under a binocular condition, the accommodative lag for each target was significantly smaller in the EOM group (analysis of variance, P<.01), but the mean slope of the accommodative stimulus-response function did not significantly differ between the EOM and EMM groups (1.05+/-0.11 and 1.02+/ 0.10 D/D, respectively). The mean slope under a binocular condition was significantly steeper than that under a monocular condition in both groups (paired t-test, P<.05). CONCLUSIONS: In adults with EOM, the accommodative stimulus-response function was not impaired, and the habitual accommodative lag was rather small, probably due to the reduced accommodative demand by a vertex distance and/or the intentional undercorrection of spectacles. PMID- 12782166 TI - Ocular factors relevant to keratoepitheliopathy in glaucoma patients with and without diabetes mellitus. AB - PURPOSE: To examine the effect of diabetes mellitus on the keratoepitheliopathy in glaucoma patients with and without diabetes mellitus who were treated with anti-glaucoma eye drops. METHODS: The presence and severity of keratoepitheliopathy was investigated in the eyes of 36 glaucoma patients with diabetes mellitus and 47 nondiabetic patients who had glaucoma. All the patients had used anti-glaucoma eye drops. The ocular factors examined were the status of the lipid layer of the tear fluid assessed by a specular reflection video recording system, the tear volume assessed by the Schirmer test, and the tear film stability assessed by tear break-up time (BUT). RESULTS: The incidence of superficial punctate keratitis (SPK) was 36.1% in the diabetic patients with glaucoma and 27.7% in the nondiabetic patients with glaucoma. Serious cases of SPK were seen only in the diabetic patients with glaucoma. The uniformity of the tear lipid layer, results of the Schirmer test, and the tear BUT in the diabetic patients with glaucoma were similar to those in the nondiabetic patients with glaucoma. CONCLUSION: In glaucoma patients who use anti-glaucoma eye drops, the effects of diabetes mellitus on the keratoepitheliopathy and other ocular factors are not significant. However, we must consider the serious cases of keratoepitheliopathy in these patients. PMID- 12782168 TI - Age-related change in contrast sensitivity among Japanese adults. AB - PURPOSE: To evaluate the age-related change in contrast sensitivity seen in a middle-aged to elderly Japanese population. METHODS: Contrast sensitivity and visual acuity were measured in subjects aged 40 to 79 years randomly recruited from a community in Aichi prefecture near Nagoya, Japan. Contrast sensitivity tests were performed using the Vistech contrast sensitivity test chart (VCTS 6500). The results were statistically analyzed relative to age. RESULTS: A statistically significant decrease in contrast sensitivity was seen with advancing age at each spatial frequency (Cochran-Mantel-Haenszel: P<.001). This trend was detected even when the subjects were limited to only those having a corrected visual acuity of 1.0 or better (Cochran-Mantel-Haenszel: P<.001). Overall, 9.4% of the eyes with good visual acuity had poor contrast sensitivity at a high spatial frequency, while in the 70-79-year-old group, the percentage with poor contrast sensitivity reached 21.1%. CONCLUSIONS: The age-related decrease in contrast sensitivity was confirmed at all frequencies in our population, even when adjusted for visual acuity. Our results suggest that contrast sensitivity tests, especially at high frequencies, assess aspects of visual function that cannot be determined in the elderly population from visual acuity tests alone. PMID- 12782169 TI - Chediak-Higashi syndrome with progressive visual loss. AB - BACKGROUND: The change of visual function in Chediak-Higashi syndrome has not been well described. CASES: The visual function of a 12-year-old Japanese girl with ocular albinism due to Chediak-Higashi syndrome was followed by periodic ophthalmological examinations. OBSERVATIONS: A lack of pigmentation in the iris and ocular fundus, and pigmentary degeneration of the peripheral retina were observed. The visual loss and the constriction of visual field progressed with increasing age. The electroretinogram was extinguished at 12 years of age. CONCLUSIONS: The constriction of the visual field may be due to the pigmentary degeneration of the ocular fundus. Ophthalmologists should be aware that progressive visual loss and the constriction of visual field can occur in patients with Chediak-Higashi syndrome as they grow older. PMID- 12782170 TI - Preoperative electroretinogram and postoperative visual outcome in patients with diabetic vitreous hemorrhage. AB - PURPOSE: To determine whether the single-flash electroretinogram (ERG) can predict the postoperative outcome in diabetic cases where massive vitreous hemorrhage precludes fundus observation. METHODS: Eighty-five diabetic patients (105 eyes) who underwent vitrectomy due to dense vitreous hemorrhage were studied retrospectively. Eyes with postoperative complications were excluded. Preoperative ERGs (mixed cone-rod ERG with maximum flash intensity) were classified as: Group A, the b-wave/a-wave ratio (b/a ratio) was > or =1.0 and the oscillatory potentials (OPs) were clearly recordable (22 eyes); Group B, the b/a ratio was > or =1.0 and the OPs were markedly reduced (33 eyes); and Group C, the b/a ratio was <1.0 (50 eyes). RESULTS: The postoperative visual acuity in Group C (hand motion to 1.5) was significantly worse than in Group A (0.4-.2) (P<.01) or Group B (0.08-1.0) (P<.01). Thick preretinal membrane causing retinal traction around the optic disc was found intraoperatively in 1 eye (4.5%) in Group A, 9 eyes (27.3%) in Group B, and 28 eyes (56.0%) in Group C (P=.0132). CONCLUSION: Our findings suggested that the configuration of the single-flash ERG can provide important preoperative information for a functional prognosis following vitrectomy in diabetic patients with vitreous hemorrhage. PMID- 12782171 TI - Choroiditis in systemic lupus erythematosus: systemic steroid therapy and focal laser treatment. AB - PURPOSE: To report a rare case of choroiditis in association with systemic lupus erythematosus (SLE). CASE: A 49-year-old woman with a 17-year history of SLE experienced acute vision impairment of her left eye during the remission stage of systemic SLE. Fundus examination revealed a gray-white subretinal exudate with serous retinal detachment. Angiographic examination disclosed choroidal inflammation at the macula and a breakdown of the blood retinal barrier. Retinal burns were applied to the subretinal exudate with an argon laser as in the treatment of central serous retinopathy. Afterward, her visual acuity showed prompt recovery due to the regression of the serous retinal detachment. However, the choroidal inflammation remained until the systemic condition was controlled with steroid therapy. RESULTS: Laser treatment of a subretinal exudate was helpful for the resolution of serous detachment and the prompt improvement of visual acuity, whereas systemic steroid therapy was effective for choroidal inflammation. CONCLUSIONS: Systemic steroid therapy is thought to be effective for SLE choroiditis; however, this therapy is also known to cause serous retinal detachment. Thus, in SLE choroiditis, laser photocoagulation at a leakage point, in addition to systemic steroid therapy, may be helpful for the prompt restoration of vision in patients with serous retinal detachment. PMID- 12782172 TI - Subconjunctival hemorrhage: the first presenting clinical feature of idiopathic thrombocytopenic purpura. AB - BACKGROUND: Subconjunctival hemorrhage as the first presenting clinical feature of idiopathic thrombocytopenic purpura, to the best of our knowledge, has not been reported earlier. CASE: A 60-year-old woman presented with an isolated finding of subconjunctival hemorrhage. She later developed hemorrhage from retinal vessels and had a single episode of hematuria. OBSERVATIONS: Her blood cell count showed an extremely low platelet count. Her medical history and clinical examination for any other systemic or ophthalmic pathology were negative. Even after treating the patient with blood and platelet transfusion and maintaining her on high doses of systemic steroids, she did not respond well. CONCLUSION: The appearance of spontaneous subconjunctival hemorrhage in a patient should be thoroughly investigated as it can be an initial sign of a grave systemic disorder. PMID- 12782179 TI - Sigma receptors: potential medications development target for anti-cocaine agents. AB - The ability of cocaine to interact with sigma receptors suggests a viable target for medications development. Recently, numerous novel compounds and antisense oligodeoxynucleotides targeting sigma receptors have been synthesized and shown to prevent the behavioral toxicity and psychomotor stimulant effects of cocaine in animals. Protective doses of sigma receptor antagonists have also been shown to prevent changes in gene expression that are induced by cocaine. Together, the studies provide insight and promising future directions for the development of potential medications for the treatment of cocaine addiction and overdose. PMID- 12782180 TI - Hydrophilic residues at position 3 highlight unforeseen features of the fMLP receptor pocket. AB - The peptides for-Met-Leu-Tyr-OMe, for-Met-Leu-Glu-OMe, for-Met-Leu-Asp-OMe and for-Met-Leu-Ser-OMe were synthesized to investigate the importance of a hydrophilic side chain of the residue at position 3 on biological activities of human neutrophils. A number of in vitro essays were carried out, including: chemotaxis, superoxide anion production, lysozyme release and receptor binding. Our results highlight that for-Met-Leu-Asp-OMe acts as a full agonist with a higher efficacy than formyl-Met-Leu-Phe-OMe, the tripeptide normally used as a model chemoattractant for the study of cell functions. The other analogs show efficacies that are in the same range or a little less than the prototype. The main point emerging from this study is that the role of Phe substitution needs to be re-hypothesised. PMID- 12782181 TI - Signalling pathways regulating inducible nitric oxide synthase expression in human kidney epithelial cells. AB - The purpose of this study was to elucidate the signalling pathways involved in the cytokine-activated inducible nitric oxide synthase (iNOS) response in a human kidney epithelial cell line, A498. Unstimulated cells did not express iNOS. Exposure of A498 cells to a cytokine mixture consisting of interferon gamma, interleukin-1 beta and tumor necrosis factor-alpha (TNF-alpha) increased nitrite production, iNOS mRNA and protein expression. Pharmacological inhibition of tyrosine kinases, including janus kinase (JAK2), and protein kinase C (PKC) inhibited cytokine-mediated nitrite production and iNOS protein expression. The involvement of mitogen-activated protein kinases (MAPKs) was investigated. Inhibition of p38 MAPK, but not of an upstream activator of extracellular signal regulated kinase (ERK), caused a decrease in iNOS expression and nitrite production in response to cytokines. Electrophoretic mobility shift assay of nuclear extract from cytokine-stimulated cells demonstrated a pronounced binding to a nuclear factor kappa B (NF-kappa B) sequence present in the human iNOS promoter. Furthermore, the NF-kappa B inhibitor pyrrolidinedithiocarbamate (PDTC) decreased cytokine-activated iNOS protein expression and nitrite production. The present study has demonstrated that cytokine-stimulated iNOS expression in human kidney epithelial cells involves activation of tyrosine kinases, including JAK2, PKC, p38 MAPK and NF-kappa B. PMID- 12782182 TI - Characterization of bradykinin-induced prostaglandin E2 release from cultured rat trigeminal ganglion neurones. AB - Bradykinin and prostaglandins are both local mediators strongly implicated in pain and inflammation. Here, we have investigated the effects of bradykinin on the release of prostaglandin E(2) from cultured neurones derived from adult rat trigeminal ganglia. Bradykinin was a potent inducer of prostaglandin E(2) release, an effect that was likely mediated by bradykinin B(2) receptors, as the bradykinin-induced prostaglandin E(2) release was attenuated by the bradykinin B(2) receptor-selective antagonist, arginyl-L-prolyl-trans-4-hydroxy-L prolylglycyl-3-(2-thienyl)-L-alanyl-L-seryl-D-1,2,3,4-tetrahydro-3 isoquinolinecarbonyl-L-(2 alpha, 3 beta, 7a beta)-octahydro-1H-indole-2-carbonyl L-arginine (HOE 140), but not by the bradykinin B(1) receptor-selective antagonist, des-Arg(9),[Leu(8)]-bradykinin. Furthermore, bradykinin-induced prostaglandin E(2) release was inhibited following treatment with the phospholipase A(2) inhibitor, arachidonyltrifluoromethyl ketone (AACOCF(3)), the nonselective cyclooxygenase inhibitor, piroxicam, the mitogen-activated protein kinase kinase-1 (MEK1) inhibitor, 2'-amino-3'-methoxyflavone (PD98059), and the protein kinase C inhibitor, bisindolylmaleimide XI (Ro320432). Taken together, these data suggest that bradykinin, acting via bradykinin B(2) receptors, induces prostaglandin E(2) release from trigeminal neurones through the protein kinase C and mitogen-activated protein kinase-dependent activation of phospholipase A(2) and consequent stimulation of cyclooxygenases. PMID- 12782183 TI - Benzodiazepine agonist and inverse agonist coupling in GABAA receptors antagonized by increased atmospheric pressure. AB - Past work found that exposure to 12 times normal atmospheric pressure (ATA) of helium-oxygen gas (heliox) selectively antagonizes (uncouples) and differentiates allosteric coupling in GABA(A) receptors initiated by benzodiazepines versus neurosteroids. The present study tested the hypothesis that pressure can differentiate coupling initiated by a spectrum of benzodiazepine receptor ligands by measuring the effects of pressure on benzodiazepine ligand modulation of GABA activated 36Cl(-) uptake in mouse brain membranes. 12 ATA completely antagonized allosteric modulation by: benzodiazepine receptor agonists diazepam and flunitrazepam; Type-1 selective benzodiazepine receptor agonist zolpidem and the benzodiazepine receptor partial inverse agonist ethyl-8-azido-5,6-dihydro-5 methyl-6-oxo-4H-imidazo[1,5-alpha][1,4]benzodiazepine-3-carboxylate (Ro15-4513). The similar, non-competitive-like characteristics of pressure antagonism of these ligands suggest common structural/functional elements underlying their coupling. Pressure also antagonized allosteric modulation by the benzodiazepine receptor inverse agonist methyl 6,7-dimethoxy-4-ethyl-beta-carboline-3-carboxylate (DMCM), but the antagonism was not complete and appeared to be surmountable (competitive like) suggesting unexpected differences in coupling for DMCM versus Ro15-4513. These studies represent the first attempt to use pressure as a tool to dissect benzodiazepine receptor coupling. The results suggest that there is a common, pressure antagonism sensitive structural/functional element underlying coupling for benzodiazepine receptor ligands and that coupling for the full inverse benzodiazepine receptor agonist DMCM differs from coupling for benzodiazepine receptor agonists and benzodiazepine receptor partial inverse agonists. PMID- 12782184 TI - Cannabinoid CB1 receptor-mediated inhibition of glutamate release from rat hippocampal synaptosomes. AB - Cannabinoid receptors are widely expressed in the brain and have been shown to regulate synaptic transmission through a presynaptic mechanism. Using synaptosomal preparation, I show here that 2,3-dihydro-5-methyl-3-(4-morpholinyl methyl)-pyrrolo-1,4-benzoxazin-6-yl-1-naphthalenylmethanone (WIN 55212-2) strongly depressed 4-aminopyridine-evoked glutamate release in a concentration dependent manner, and this effect was reversed by the selective cannabinoid CB(1) receptor antagonist 1-(2,4-dichlorophenyl)-5-(4-iodophenyl)-4-methyl-N-4 morpholinyl-1H-pyrazole-3-carboxamide (AM 281). The inhibitory modulation by WIN 55212-2 was not due to a decrease in synaptosomal excitability or a direct effect on the release machinery because WIN 55212-2 did not alter 4-aminopyridine mediated depolarization and ionomycin-induced glutamate release. In addition, the WIN 55212-2-mediated inhibition of glutamate release was blocked by the G(i)/G(o) protein inhibitor pertussis toxin, but not by the protein kinase A inhibitor 2,3,9,10,11,12-Hexahydro-10-hydroxy-9-methyl-1-oxo-9,12-epoxy-1H-diindolo benzodiazocine-10-carboxylic acid, hexyl ester (KT 5720). Furthermore, this inhibitory effect was associated with a decrease in 4-aminopyridine-evoked Ca(2+) influx, which could be completely prevented in synaptosomes pretreated with the N and P/Q-type Ca(2+) channel blockers. Together, these observations indicate that activation of cannabinoid CB(1) receptors inhibit 4-aminopyridie-evoked glutamate release from hippocampal synaptosomes through a inhibitory G protein to suppress N- and P/Q-type Ca(2+) channel activity. PMID- 12782185 TI - Activation of peripheral ATP-sensitive K+ channels mediates the antinociceptive effect of Crotalus durissus terrificus snake venom. AB - The role of peripheral potassium channels on the antinociceptive effect of Crotalus durissus terrificus venom, a mixed delta- and kappa-opioid receptor agonist, was investigated in hyperalgesia induced by carrageenin or prostaglandin E(2). Rat paw pressure test was applied before and 3 h after the intraplantar (i.pl.) injection of the nociceptive stimuli. Oral administration of venom 2 h after carrageenin or prostaglandin E(2) induces antinociception. Local pretreatment with 4-aminopyridine and tetraethylammonium (blockers of voltage dependent K(+) channel) or charybdotoxin and apamin (inhibitors of large- and small-conductance Ca(2+)-activated K(+) channel, respectively) did not modify venom effect. On the other hand, glybenclamide, an inhibitor of ATP-sensitive K(+) channel abolished antinociception induced by the venom. Glybenclamide also inhibited the antinociceptive effect of [D-Pen(2.5)] enkephalin (DPDPE), a delta opioid receptor agonist, but did not modify the effect of (+)-trans-(1R,2R)-U 50488 (U50488), a kappa opioid receptor agonist. Diazoxide and pinacidil, two ATP sensitive K(+) channel openers, injected by intraplantar route, induced a long lasting increment of pain threshold of the animals and produced antinociception in both models of hyperalgesia. These results suggest that the antinociceptive effect of crotalid venom is mediated by activation of ATP-sensitive K(+) channels at peripheral afferent neurons. PMID- 12782186 TI - Does Mg2+ deficiency induce a long-term sensitization of the central nociceptive pathways? AB - In rats, a Mg(2+)-deficient diet, which in a few days dramatically decreased the Mg(2+) concentration in plasma, cerebrospinal fluid (CSF) and spinal cord, was accompanied by a significant lowering of the nociceptive threshold. After reloading, the Mg(2+) concentration was rapidly normalized in both spinal cord and CSF. In parallel, the neurological disturbances induced by Mg(2+) deficiency vanished in less that 24 h, but the reversal of the hyperalgesia was delayed for up to 11 to 20 days. In this model, repeated doses of dizocilpine (MK-801), a non competitive NMDA receptor antagonist, given at start of the Mg(2+)-depleted diet, prevented hyperalgesia, suggesting the involvement of NMDA receptor channels. The delayed recovery of a normal pain threshold argues for long-term sensitization of the nociceptive pathways. PMID- 12782187 TI - Morphine and dipyrone co-administration delays tolerance development and potentiates antinociception. AB - This work analyses the time course of tolerance development and antinociceptive potentiation throughout repeated co-administration of morphine (an opioid receptor agonist) plus dipyrone (a non-steroidal anti-inflammatory drug) in the tail-flick test. Male Wistar rats were i.v. injected with morphine (3.1 mg/kg), dipyrone (600 mg/kg) or the combination morphine/dipyrone twice a day for 5 days. Dipyrone produced antinociceptive effects with a trend towards tolerance development at the end of the treatment. Morphine was initially effective, but complete tolerance developed after its fifth administration. The combination of morphine plus dipyrone produced a significant potentiation and longer duration of antinociceptive effects. The antinociceptive efficacy of morphine and dipyrone co administration gradually decreased after the sixth injection. An additional group of rats treated with dipyrone for 11 days developed complete tolerance after the 19th administration. These data suggest that repeated co-administration of morphine plus dipyrone results in a delay of tolerance development and in a potentiation of their individual antinociceptive effects. PMID- 12782188 TI - Antidepressant-like effect of agmatine and its possible mechanism. AB - In mammalian brain, agmatine is an endogenous neurotransmitter and/or neuromodulator, which is considered as an endogenous ligand for imidazoline receptors. In this study, the antidepressant-like action of agmatine administered p.o. or s.c. was evaluated in three behavioral models in mice or rats. Agmatine at doses 40 and 80 mg/kg (p.o.) reduced immobility time in the tail suspension test and forced swim test in mice or at dose 20 mg/kg (s.c.) in the forced swim test. Agmatine also reduced immobility time at 10 mg/kg (p.o.) or at 1.25, 2.5 and 5 mg/kg (s.c.) in the forced swim test in rats. These results firstly indicated that agmatine possessed an antidepressant-like action. With 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and lactic dehydrogenase (LDH) assay, 1, 10 and 100 microM agmatine or a classical antidepressant, 2.5 and 10 microM desipramine, protected PC12 cells from the lesion induced by 300 microM N-methyl-D-aspartate (NMDA) treatment for 24 h. Using high-performance liquid chromatography with electrochemical detection (HPLC ECD), it was found that the levels of monoamines including norepinephrine, epinephrine, dopamine or 5-hydroxytryptamine (5-HT) in PC12 cells decreased after the treatment with 200 microM NMDA for 24 h, while in the presence of 1 and 10 microM agmatine or 1 and 5 microM desipramine, the levels of norepinephrine, epinephrine or dopamine were elevated significantly while 5-HT did not change. Moreover, norepinephrine, 5-HT or dopamine had the same cytoprotective effect as agmatine at doses 0.1, 1 and 10 microM. In the fura-2/AM (acetoxymethyl ester) labeling assay, 1 and 10 microM agmatine, 1 and 5 microM desipramine or monoamines norepinephrine, 5-HT at doses 0.1 and 1 microM attenuated the intracellular Ca(2+) overloading induced by 200 microM NMDA treatment for 24 h in PC12 cells. In summary, we firstly demonstrated that agmatine has an antidepressant-like effect in mice and rats. A classical antidepressant, desipramine, as well as agmatine or monoamines protect the PC12 cells from the lesion induced by NMDA treatment. Agmatine reverses the NMDA-induced intracellular Ca(2+) overloading and the decrease of monoamines (including norepinephrine, epinephrine or dopamine) contents in PC12 cells, indicating that agmatine's antidepressant-like action may be related to its modulation of NMDA receptor activity and/or reversal of the decrease of monoamine contents and Ca(2+) overloading induced by NMDA. PMID- 12782189 TI - Antinociception by a peripherally administered novel endomorphin-1 analogue containing beta-proline. AB - We previously described a novel endomorphin-1 analogue (Tyr-L-beta-Pro-Trp-Phe NH(2); Endo1-beta-Pro) more resistant to enzymatic hydrolysis than endomorphin-1 that acts as a mu-opioid receptor agonist. In this study we report that Endo1 beta-Pro, s.c. injected in the mouse, is an effective antinociceptive agent in the tail flick (ED(50)=9.2 mg/kg) and acetic acid-induced abdominal constriction (ED(50)=1.2 mg/kg) tests. Moreover, s.c. Endo1-beta-Pro significantly decreases, in the mouse, the gastrointestinal propulsion measured as transit of an orally administered charcoal meal (ED(50)=10.0 mg/kg). Subcutaneous beta-funaltrexamine or a high dose of the mu(1)-opioid receptor-selective antagonist naloxonazine (50 mg/kg) prevents the antinociceptive and antitransit action of Endo1-beta-Pro; moreover, these effects are partially blocked by i.c.v. naloxone or by i.p. naloxone methiodide, this latter does not readily cross the blood-brain barrier. On the contrary, the kappa-opioid receptor antagonist nor-binaltorphimine or the delta-opioid receptor antagonist naltrindole are ineffective Thus, Endo1-beta-Pro may act, preferentially, through central and peripheral mu(2)-opioid receptors to produce antinociception and to inhibit gastrointestinal transit. Endo1-beta-Pro is among the first endomorphin-1 analogues showing antinociceptive activity after systemic administration. This compound will be extremely useful for exploring the pharmacological profile of endomorphins in vivo and confirms the potential therapeutic interest of endomorphin derivatives as novel analgesic agents. PMID- 12782190 TI - Anxiolytic profile of the antiepileptic drug levetiracetam in the Vogel conflict test in the rat. AB - The novel antiepileptic drug levetiracetam has been shown to reverse anxiogenic effects of benzodiazepine withdrawal in mice tested in an elevated plus-maze without altering the behaviour of normal mice in this model. This could suggest that the effect of levetiracetam is dependent upon the level of stress/anxiety of the animals. Levetiracetam was therefore further examined in another widely used animal model of anxiety, the Vogel conflict test. In the first experiment, water deprived rats were submitted to a free drinking period (habituation) in a chamber equipped with a bottle of water. Twenty-four hours later, animals were returned to the same chamber but the licks to the water bottle were then punished with a foot shock (0.5 mA, 90 ms). In the second experiment, the procedure was modified by administering a foot shock at the end of the habituation period in order to induce a state of stress/anxiety (conditioned fear/ anticipatory anxiety) for subsequent testing. Levetiracetam (17 and 54 mg/kg) and chlordiazepoxide (5 mg/kg) were administered via the intraperitoneal route. The results indicated that in the first experiment only chlordiazepoxide showed a statistically significant anxiolytic effect. In contrast, in the second experiment, where the shock was given at the end of the habituation period, levetiracetam (54 mg/kg) revealed significant anxiolytic activity similar to chlordiazepoxide. This suggests that levetiracetam may have potential anxiolytic effects and may provide therapeutic benefits to individual with anxiety spectrum disorders. PMID- 12782191 TI - Long-lasting behavioral changes induced by pre- or neonatal exposure to diazepam in rats. AB - Prenatal treatment with small doses of diazepam may counteract the effect of physical stress in rats, normalizing the time course of neonatal reflexes and the behavioral responses in adulthood. However, prenatal administration of diazepam in greater doses may induce desensitization of gamma-aminobutyric acid A (GABA(A)) receptors and induce hypersensitivity to convulsants. This study was designed to examine in rats the influence of prenatal or neonatal diazepam treatment on development of neonatal reflexes, as index of brain maturation, and susceptibility to pentylenetetrazol-induced convulsions in adulthood. Pregnant Wistar rats were injected with diazepam 2.5 mg/kg/day, intraperitoneally (i.p.) on days 14-21 of pregnancy. Offspring showed a delay in the appearing of neonatal reflexes (cliff aversion, forelimb placing, forelimb grasping and bar holding) except for righting and startle reflexes. At 50 days of age, male rats showed a greater sensitivity to pentylenetetrazol compared to controls. In contrast, females showed a decreased susceptibility to convulsions. The appearance of reflexes in pups exposed to diazepam during neonatal life appeared to be similar to that of controls. Only the appearance of cliff aversion and startle reflexes appeared to be delayed in animals exposed neonatally to diazepam as compared to controls. In adulthood, female rats exposed in early neonatal life to diazepam again showed a resistance to pentylenetetrazol-induced convulsions as compared to male animals. These data suggest that prenatal exposure to diazepam induces long lasting behavioral changes, which may be influenced by sex-dependent factors. PMID- 12782192 TI - Effects of a selective agonist and antagonist of CRF2 receptors on cardiovascular function in the rat. AB - The aim of the present study was to investigate the effects of activation or blockade of the CRF(2) receptor subtype on cardiovascular function in conscious rats following systemic i.v. administration of the CRF(2) receptor peptide agonist urocortin 2 given alone and the selective CRF(2) receptor peptide antagonist antisauvagine-30 given alone. Urocortin 2 caused a dose-dependent reduction in mean arterial blood pressure and a dose-dependent increase in heart rate. Pretreatment with antisauvagine-30 blocked the hypotensive effect of urocortin 2. Antisauvagine-30 failed to produce any statistically significant effects on mean arterial blood pressure and heart rate at doses that completely blocked the effects of urocortin 2. These data verify the cardiovascular effects of selective CRF(2) receptor activation, but find no evidence for an endogenous CRF(2)-mediated tone. PMID- 12782193 TI - Pre- and postsynaptic inhibitory potencies of the angiotensin AT1 receptor antagonists eprosartan and candesartan. AB - The aim of the present study was to determine the inhibitory potency of two selective angiotensin AT(1) receptor antagonists, eprosartan and candesartan, at the level of the sympathetic nerve terminal and the vascular smooth muscle. Male New Zealand White rabbits, weighing 2100-2550 g, were used. To study eprosartan and candesartan at the neuronal angiotensin AT(1) receptor, we investigated their influence on the angiotensin II-enhanced, electrical field stimulation-evoked sympathetic transmission in the rabbit isolated thoracic aorta in a noradrenaline spillover model. To study both antagonists at the vascular angiotensin AT(1) receptor, concentration-response curves for angiotensin II were constructed in the presence or absence of the two angiotensin AT(1) receptor antagonists. Angiotensin II (10 nM) caused a significant increase by 107+/-11.1% of the stimulation-evoked sympathetic outflow, which was concentration-dependently inhibited by both eprosartan (pIC(50) 7.91+/-0.12) and candesartan (pIC(50) 10.76+/-0.13). Angiotensin II (1 nM-0.3 microM) caused a concentration-dependent increase in contractile force (E(max) 20.62+/-2.24 mN, pD(2) 8.16+/-0.04). Both eprosartan (pA(2) 8.90+/-0.11, pIC(50) 8.87+/-0.12 (10 nM angiotensin II)) and candesartan (pD(2)' 10.80+/-0.13) counteracted the contractions evoked by cumulative concentrations of angiotensin II. Candesartan proved a more potent antagonist than eprosartan at both the pre- and postjunctional angiotensin AT(1) receptor. For eprosartan, vascular inhibitory concentrations were 10-fold lower than sympatho-inhibitory concentrations, whereas for candesartan, inhibitory concentrations at both sites were similar. The results may be explained by differences between the pre- and postjunctional angiotensin AT(1) receptor subtype. PMID- 12782194 TI - S1P3 receptors mediate the potent constriction of cerebral arteries by sphingosine-1-phosphate. AB - We characterized the effect of Sphingosine-1-phosphate (S1P) on vascular tone. S1P selectively constricted isolated cerebral, but not peripheral arteries, despite ubiquitous expression of S1P(1), S1P(2), S1P(3) and S1P(5) receptor mRNA. Clostridium B and C3 toxins and the rho-kinase inhibitor Y27632 (trans-N-(4 pyridyl)-4-(l-aminoethyl)-cyclohexane carboxamide) reduced this vasoconstriction to S1P, indicating that the response was mediated through Rho. Pertussis toxin displayed only weak inhibition, suggesting minor involvement of G(i/o) protein. The S1P effect was specifically reduced by adenovirus bearing a s1p(3) but not s1p(2), antisense construct. Furthermore, suramin, which selectively blocks S1P(3) receptors, inhibited the vasoconstrictor effect of S1P, indicating that S1P(3) receptors account for at least part of S1P-mediated vasoconstriction in cerebral arteries. In vivo, intracarotid injection of S1P decreased cerebral blood flow, an effect prevented by suramin treatment. Because S1P constricts cerebral blood vessels and is released from platelets during clotting, the S1P/S1P(3) system constitutes a novel potential target for cerebrovascular disease therapy. PMID- 12782195 TI - Pregnancy-associated changes in responsiveness of the porcine myometrium to bioactive substances. AB - To determine the pregnancy-associated changes in the porcine uterine contractility, the spontaneous contraction and the mechanical responses to bioactive substances of uteri in nonpregnant proestrus and pregnant pigs (25-60 days of gestation) were compared in vitro. Longitudinal muscle (LM) and circular muscle (CM) of the uterus exhibited spontaneous contraction, but the frequency in pregnant pigs was lower than that in the nonpregnant pigs. The duration and force of spontaneous contraction in the pregnant pigs were long and large compared with both in the nonpregnant pigs. L-Nitroarginine methylester (L-NAME) and 2a-[4-(4 phenyl-1,2,3,6-tetrahydropyridyl)butyl]-2a,3,4,5-tetrahydro-benzo[cd]indol-2(1H) one (DR4004) did not change the spontaneous contraction in the uteri of nonpregnant pigs but increased its amplitude in the uteri of pregnant pigs. Isoprenaline inhibited the uterine spontaneous contraction of the nonpregnant and pregnant pigs, and the inhibition was stronger in the pregnant than in the nonpregnant pigs. 5-Hydroxytryptamine also caused inhibition of spontaneous contraction in the uteri of nonpregnant pigs (CM>LM). In the pregnant pigs, sensitivity to 5-hydroxytryptamine increased in a muscle layer-dependent manner (LM>CM) and difference in the responsiveness between LM and CM decreased. Acetylcholine contracted the uterine LM and CM strips of the pregnant and nonpregnant pigs. The responsiveness of CM increased slightly during pregnancy, but that of the LM did not change. 5-Bromo-N-(2-imidazolin-2-yl)-6 quinoxalinamine (UK14304) caused contraction of only LM in the uteri of nonpregnant pigs, but contracted both LM and CM strips in the pregnant pigs. Oxytocin and prostaglandin F(2 alpha) also contracted the uteri of nonpregnant pigs (LM>CM). Pregnancy increased the contraction of both agents in the LM and CM, but the increment was marked in the CM. The contractile forces induced by all stimulants were increased (by 1.7- to 2.5-fold) in the LM and CM of pregnant pigs. In conclusion, (1) low frequency, slow kinetics and large force of spontaneous contraction are characteristics of the pregnant porcine uteri, and nitric oxide and 5-hydroxytryptamine are supposed to be partially involved in the regulation of spontaneous contraction, and (2) responses to both contractile and inhibitory agents are increased in the pregnant pigs. Increment of the responsiveness is conspicuous in the muscle layer that is less sensitive to each agonist in the uteri of nonpregnant pigs. According to the pregnancy-associated changes, muscle layer-related differences of responsiveness to bioactive substances in the nonpregnant pigs tend to decrease in the pregnant pigs. PMID- 12782196 TI - Protective effect of melatonin against mitochondrial injury induced by ischemia and reperfusion of rat liver. AB - Melatonin, a pineal secretory product, is a potent scavenger of a variety of free radicals. The aim of this study was to investigate the effect of melatonin on the prevention of mitochondrial injury induced by hepatic ischemia and reperfusion. Rats were subjected to 70 min of hepatic ischemia and 2 h of reperfusion. Fifteen minutes prior to ischemia and at reperfusion, animals received vehicle or melatonin (10 mg/kg body weight) intraperitoneally. In the vehicle-treated animals, the respiratory control index, ADP/O, State 3 respiration and dinitrophenol-induced uncoupled respiration decreased markedly after ischemia/reperfusion and were restored by melatonin administration. Similarly, pH change coupled with mitochondrial energy transfer was suppressed by ischemia/reperfusion with the effects being reduced by melatonin treatment. Mitochondrial lipid peroxidation was elevated in the ischemic/reperfused vehicle treated livers, but this elevation was attenuated by melatonin. Mitochondrial glutathione peroxidase activity decreased in the vehicle-treated group with this decrease being reduced by melatonin treatment. Electron microscopic studies demonstrated that treatment with melatonin restored to near normal the ischemia/reperfusion-induced disorganization of mitochondrial structure. Melatonin protects against mitochondrial injury which reduces mitochondrial oxidative stress and improves ischemia/reperfusion-induced hepatic energy metabolism. PMID- 12782197 TI - Streptozotocin-induced diabetes modulates presynaptic and postsynaptic function in the rat ileum. AB - Altered gastrointestinal motility frequently occurs in diabetic patients and also in animal models of diabetes but the underlying causes are not clear. In the present study, contractile responses to agonists and electrical field stimulation (EFS) and the inhibitory actions of an adenosine A(1) receptor agonist were investigated on ilea from 8-week streptozotocin (STZ)-induced diabetic rats. Contractile responses to carbachol, prostaglandin F(2 alpha) (PGF(2 alpha)), the calcium ionophore A23187 and to EFS were increased in diabetic tissues compared to controls. In contrast, the inhibitory effects of a potent and selective adenosine A(1) receptor agonist N(6)-cyclopentyladenosine (CPA) on electrical field stimulation-evoked contractions were decreased in diabetic tissues compared to controls but its ability to relax carbachol-contracted tissues was unaltered. These results suggest that diabetes may cause alterations at both pre- and postsynaptic sites and this may lead in turn to the gastrointestinal complications seen in diabetic patients. PMID- 12782198 TI - Antitussive principles of Glycyrrhizae radix, a main component of the Kampo preparations Bakumondo-to (Mai-men-dong-tang). AB - We attempted to elucidate the antitussive principles of Glycyrrhizae radix, a main component of Bakumondo-to (Mai-men-dong-tang). Although the 50% methanol eluted fraction (100 mg/kg, p.o.) caused a more than 60% reduction in the number of capsaicin-induced coughs, neither the water-eluted nor 100% ethanol-eluted fractions of water extract of G. radix had antitussive effects. The water extract of G. radix contained high levels of liquiritin, liquiritin apioside, isoliquiritin, isoliquiritin apioside and glycyrrhizin. On the other hand, the 50% methanol-eluted fraction contained mainly liquiritin and liquiritin apioside, but not the other compounds. Liquiritin apioside (3-30 mg/kg, p.o.), but not liquiritin, isoliquiritin, isoliquiritin apioside or glycyrrhizin, dose dependently inhibited the number of coughs. Methysergide, a serotonin receptor antagonist, antagonized the antitussive effect of liquiritin apioside. However, the antitussive effect of liquiritin apioside was not antagonized by naloxone. Pretreatment with glibenclamide (3 mg/kg, i.p.), an ATP-sensitive potassium channel blocker, also significantly reduced the antinociceptive effect of liquiritin apioside. These results suggest that G. radix contains a potent antitussive compound, liquilitin apioside, whose antitussive effect may depend on both peripheral and central mechanisms. PMID- 12782199 TI - Amiodarone minimizes experimental autoimmune myocarditis in rats. AB - Amiodarone, a promising drug for the treatment of tachyarrythmias, was recently found to have immunomodulatory effects in vitro. We hypothesized that amiodarone would affect the immune system in vivo and examined the effect of amiodarone on myocarditis in rats. We induced experimental autoimmune myocarditis in rats by cardiac myosin immunization and treated the animals with an intraperitoneal injection of amiodarone at 25 mg/kg/every other day, 10 times after the induction of experimental autoimmune myocarditis. In the treated group, both microscopic and macroscopic examinations showed reduced heart weights, a mild and localized infiltration of inflammatory cells and fibrosis in the myocardium, and a mild congestion in the liver and lungs as compared with the control group. The phenotypic distribution of lymphocytes in peripheral blood showed a significant decrease in the CD4/CD8a ratio in the treated group, but not in the control group. The proportion of mast cells involved in inflammatory cell infiltration was lower in the treated group than the control group. In vitro, amiodarone inhibited the proliferation of mast cells by arresting them in the G2 phase of the cell cycle. These results indicated that amiodarone minimized the progression of experimental autoimmune myocarditis, suggesting a potential therapeutic role for amiodarone treatment in patients suffering from myocarditis, especially myocarditis complicated by cardiac arrhythmias. One possible mechanism by which amiodarone minimizes the progression of experimental autoimmune myocarditis may be to affect the immune system via the immunomodulatory effects on T cell and mast cell functions. PMID- 12782200 TI - Intact noradrenaline transporter is needed for the sympathetic fine-tuning of cytokine balance. AB - Earlier studies demonstrated that cytokine production is under the tonic control of noradrenaline. As the level and/or the duration of noradrenaline action is regulated by the noradrenaline transporter (NET), which is also a target of antidepressant treatment, we studied its role in the regulation of the cytokine response during inflammation. The endotoxin-evoked tumour necrosis factor-alpha (TNF-alpha) and interleukin-10 response was studied in genetically produced noradrenaline transporter-deficient (NET-KO) mice and by treatment with desipramine, a monoamine uptake-blocker antidepressant. NET-KO mice responded to endotoxin with significantly lower TNF-alpha and interleukin-10 production in comparison to their wild-type counterparts. Functional involvement of both alpha- and beta-adrenoceptors could be demonstrated in our model systems, using 7,8 methylenedioxy-14 alpha-hydroxy-alloberbane.HCl (CH-38083) and propranolol; however, the differences between the two phenotypes remained, suggesting a limited role of alpha-adrenoceptors in the observed changes. Acute treatment of both wild-type and NET-KO mice with desipramine significantly decreased the TNF alpha response and significantly increased interleukin-10 production, indicating the role of an intact noradrenaline transporter in anti-inflammatory responses. PMID- 12782201 TI - Inhibitors of poly (ADP-ribose) polymerase modulate signal transduction pathways in colitis. AB - During inflammatory bowel diseases, oxidative and nitrosative stress induces DNA damage and activation of the nuclear enzyme poly (ADP-ribose) polymerase (PARP), resulting in depletion of intracellular energetics, intestinal barrier dysfunction and cellular death. The aim of our study was to evaluate the therapeutic efficacy of in vivo inhibition of PARP in experimental colitis, which was induced by rectal instillation of trinitrobenzene sulfonic acid (TNBS) in rats. In vehicle-treated rats, TNBS treatment resulted in colonic erosion and ulceration. Neutrophil infiltration (indicated by myeloperoxidase activity in the colon) was associated with formation of nitrotyrosine and marked apoptosis. Elevated levels of plasma nitrate/nitrite, metabolites of nitric oxide (NO), were also found. These inflammatory events were associated with the activation of nuclear factor-kappa B (NF-kappa B) and activator protein-1 (AP-1) in the colon; NF-kappa B was maximally activated at 3 and 7 days, whereas AP-1 increased 1 day after TNBS administration and declined thereafter. Treatment of the rats with the PARP inhibitors, 3-aminobenzamide or 1,5-dihydroxyisoquinoline, resolved colonic damage and reduced plasma levels of NO metabolites. Resolution of the damage was associated with reduction of neutrophil infiltration, nitrotyrosine formation and apoptosis. Treatment with PARP inhibitors also reduced DNA binding of NF-kappa B and AP-1 in the colon. These data demonstrate that pharmacological inhibition of PARP ameliorates colitis. Reduction of the inflammatory process is associated with modification of the activation of signal transduction pathways. PMID- 12782202 TI - Interrelationship among insulin, glucagon and somatostatin secretory responses to exendin-4 in the perfused rat pancreas. AB - We have investigated the effect of exendin-4 on insulin, glucagon and somatostatin output in the perfused rat pancreas. At 9 mM glucose, exendin-4 potentiated the insulin and somatostatin responses to arginine and reduced the glucagon response to this amino acid. Thus, this reduction might be thought to be paracrine-mediated through the concomitant increase in insulin and somatostatin concentrations. At 3.2 mM glucose, exendin-4 did not affect insulin secretion, reduced glucagon release and stimulated somatostatin output. Furthermore, exendin 4 reduced glucagon secretion as induced by a glucose decline (from 11 to 3.2 mM) without affecting insulin or somatostatin responses. In summary, exendin-4 stimulated insulin and somatostatin secretion and reduced glucagon release. The glucagonostatic effect of exendin-4 was observed under conditions in which insulin and somatostatin were not affected, thus indicating that exendin-4, per se, inhibits A-cell secretion. Indeed, an additional glucagonostatic effect of exendin-4, mediated by its stimulation of insulin and/or somatostatin secretion, cannot be ruled out. PMID- 12782204 TI - Modulation by sudden darkness of apomorphine-induced behavioral responses. AB - Sudden darkness increases motor activity and decreases anxiety. In the present study, we focused on the role of dopaminergic mechanisms involved in the effects of sudden darkness. The influence of sudden darkness on the behavioral effects of low (0.05 and 0.1 mg/kg) and high (0.25, 0.45 and 0.6 mg/kg) doses of apomorphine (APO) was tested. We assayed the effects of low APO doses on yawning-penile erection syndrome (YES; 0.05 and 0.1 mg/kg) and on motor activity (0.05 mg/kg), and the effects of high APO doses on motor activity (0.25 mg/kg) and stereotyped behavior (0.45 mg/kg and 0.6 mg/kg). Spontaneous total and genital grooming of male and female rats were also recorded. Sudden darkness modified some spontaneous behaviors and also modulated several APO-induced behavioral effects. It increased spontaneous total grooming and genital grooming in male rats but had no effect on these parameters in female rats. These results show sexual dimorphism for total and genital grooming in both control and sudden darkness conditions. APO was able to induce YES in a dose-dependent manner. Sudden darkness decreased yawning elicited by both 0.05 and 0.1 mg/kg of the drug. No other parameter of YES was modified. In the open-field test, sudden darkness increased total locomotion and rearing and decreased immobility duration. APO at a dose of 0.05 mg/kg had the opposite effect on these parameters under light conditions; none of them were modified by sudden darkness. Animals treated with APO at 0.25 mg/kg, a dose that augmented total locomotion and rearing and diminished immobility duration, were clearly divided into two groups according to their responses, i.e., hypo- and hyper-responsive rats. Sudden darkness improved total locomotion and rearing, reduced immobility duration and total grooming in the hyporesponsive group, and showed no effects on the hyper-responsive group. Sudden darkness caused no modifications of stereotyped behavior. These results may be due to a sudden darkness-induced physiological release of dopamine that diminishes pre-synaptic responses to APO and increases low-intensity post synaptic responses such as motor activity without modifying high-intensity post synaptic responses such as stereotyped behavior. PMID- 12782203 TI - Decreased hypothalamic concentration of neuropeptide Y correlates with onset of hyperphagia in fa/fa rats on postnatal day 12. AB - An increased action of hypothalamic neuropeptide Y (NPY) has been proposed as a major factor in the pathophysiology of the obesity syndrome in Zucker (fa/fa) rats. Using a developmental strategy to test this hypothesis, we showed previously that significantly more arcuate NPY was expressed in fa/fa pups than in lean littermates on postnatal day (P) 2 and throughout the preweaning period [Physiol. Behav. 67 (1999) 521], and that hyperphagia first appeared on P12 [Am. J. Physiol. 275 (1998) R1106]. To test the hypothesis further, we used a specific radioimmunoassay to measure the concentration of hypothalamic NPY peptide in lean (+/+ and +/fa) and obese fa/fa Zucker rat pups on P9, P10, and P12. The concentration of NPY in fa/fa pups was not significantly different from that of the other genotypes. There was, however, a significant decrease in NPY concentration from P9 to P12 in fa/fa pups, but not in lean pups. The combination of increased NPY message and decreasing concentration of NPY peptide in fa/fa pups with age is consistent with, but does not prove, increased release of hypothalamic NPY in fa/fa pups just before and on P12 when hyperphagia emerges. These results provide further support for the importance of hypothalamic NPY in the phenotypic expression of hyperphagia in the fa/fa pups during the second postnatal week. PMID- 12782205 TI - Effects of glucose on responsiveness to change in young adult and middle-aged rats. AB - After a 0.5-, 15-, or 30-min intertrial interval, first entries of a novel Y-maze arm that had changed in brightness, percent entries of the arm, and percent time spent in this arm during a 1-min period were recorded in 4-month-old hooded rats following 6 or 30 min of free exploration of both arms. From the results, it was concluded that maximal responsiveness to the changed (or novel) arm occurred after 6 min of exploration and a 30-min intertrial interval. In a second experiment, responsiveness to change was assessed in young adult (4 months) and middle-aged (18 months) rats after 6 min of free exploration followed by an intraperitoneal injection of distilled water, or 50 or 100 mg/kg d-glucose before testing 20 min later. While glucose increased first entries of the changed arm in all rats, longer-term responsiveness in the form of percent entries of the novel arm and time spent in the novel arm was increased only for young adults. Although the results suggested age-specific glucose-enhanced consolidation or retrieval of change-related information, it was also possible that the treatment had differentially increased preferences for novelty in the two age groups. This possibility should be addressed in future research. PMID- 12782206 TI - Changes in peripheral energy metabolism during audiogenic seizures in rats. AB - Plasma glucose and lactate, hepatic glycogen and epididymal adipose tissue lipogenesis and lipolysis were studied in Wistar audiogenic rats (WARs), a genetic model of epilepsy, under three experimental conditions, i.e., before, 3 min after and 10 min after seizures induced by intense sound exposure. Plasma glucose increased 3 min after the seizure onset and rose to a peak after 10 min. Hepatic glycogen decreased significantly in susceptible audiogenic rats compared to nonepileptic controls, even before sound stimulation. A marked ( approximately 10-fold) rise was observed in plasma lactate levels 3 and 10 min after the seizures compared to the response of the control group. Lipogenic activity showed a marked decrease even after stimulation with 25 ng/ml insulin. Based on these results, WARs showed reduced isoproterenol-stimulated lipolysis compared to control animals, whereas basal levels only differed significantly at 10 min after seizure activity. In conclusion, it can be inferred from these results that (a) the increase in plasma glucose after stimulation might result from sequential interaction of autonomic activation at seizure onset; (b) excessive muscular activity was at least partially responsible for the steady rise in plasma lactate concentrations; (c) audiogenic seizures, which increase adrenergic activity, induce desensitization of the beta-adrenergic lipolytic pathway in epididymal adipose tissue; (d) genetic selection for audiogenic seizure susceptibility results in pronounced alterations at multiple levels of metabolic regulation. PMID- 12782207 TI - Leptin resistance in mice is determined by gender and duration of exposure to high-fat diet. AB - Mice fed a high-fat diet are reported to be resistant to peripheral injections of leptin. We previously failed to induce leptin resistance in female mice fed a high-fat diet for 15 weeks. Therefore, we measured the responsiveness to peripheral infusions (10 microg/day) of leptin, and the responsiveness to third ventricle injections of leptin (1 microg) in male and female NIH Swiss mice fed low-fat (10% kcal) or high-fat (45% kcal) diets. Male and female 15-week-old mice that had been fed low- or high-fat diet from 10 days of age lost fat during a 13 day intraperitoneal infusion of leptin and lost weight in response to a single central injection of leptin. Fifteen-week-old male mice fed a high-fat diet for 5 weeks did not lose body fat during a peripheral infusion of leptin and did not lose weight in response to a central injection of leptin. Female mice fed high fat diet for 5 weeks remained leptin-responsive. Weight loss was achieved without a significant voluntary decrease in food intake, suggesting that both peripherally and centrally administered leptin increases energy expenditure. These results demonstrate that the development of leptin resistance in NIH Swiss mice fed a high-fat diet is dependent upon the gender of the mice and either the duration of exposure to high-fat diet or the age at which the mice are first exposed to the diet. PMID- 12782208 TI - Physiological mechanisms mediating aspartame-induced satiety. AB - Aspartame has been previously shown to increase satiety. This study aimed to investigate a possible role for the satiety hormones cholecystokinin (CCK) and glucagon-like peptide-1 (GLP-1) in this effect. The effects of the constituents of aspartame, phenylalanine and aspartic acid, were also examined. Six subjects consumed an encapsulated preload consisting of either 400 mg aspartame, 176 mg aspartic acid+224 mg phenylalanine, or 400 mg corn flour (control), with 1.5 g paracetamol dissolved in 450 ml water to measure gastric emptying. A 1983-kJ liquid meal was consumed 60 min later. Plasma CCK, GLP-1, glucose-dependent insulinotropic polypeptide (GIP), glucose, and insulin were measured over 0-120 min. Gastric emptying was measured from 0 to 60 min. Plasma GLP-1 concentrations decreased following the liquid meal (60-120 min) after both the aspartame and amino acids preloads (control, 2096.9 pmol/l min; aspartame, 536.6 pmol/l min; amino acids, 861.8 pmol/l min; incremental area under the curve [AUC] 60-120 min, P<.05). Desire to eat was reduced from 60 to 120 min following the amino acids preload (control, -337.1 mm min; aspartame, -505.4 mm min; amino acids, -1497.1 mm min; incremental AUC 60-120 min, P<.05). However, gastric emptying rates, plasma CCK, GIP, insulin, and glucose concentrations were unaffected. There was a correlation between the increase in plasma phenylalanine and decrease in desire to eat after the liquid meal following the constituent amino acids (r=-.9774, P=.004). In conclusion, it is unlikely that aspartame increases satiety via CCK- or GLP-1-mediated mechanisms, but small changes in circulating phenylalanine concentrations may influence appetite. PMID- 12782209 TI - Excessive weight gains in female rats with transections of the stria terminalis. AB - Recent studies have demonstrated hyperphagia and excessive weight gains in female rats with small lesions in the most posterodorsal aspects of the medial amygdala. In the present study, similar results were observed in female rats with bilateral transections of the stria terminalis just as it exits the amygdala to begin its dorsal ascent (mean weight gain of 35.9 g/20 days compared to 0.1 g/20 days for operated control animals). Cellular damage caused by the retractable wire knife was limited to the caudal globus pallidus. The results of previous studies that failed to observe weight gains after stria terminalis transections were attributed to the use of male animals. The present results, along with the pattern of anterograde degeneration that is observed after obesity-inducing amygdaloid lesions, suggest a medial amygdala-stria terminalis-medial hypothalamic pathway in the regulation of food intake and body weight, but other possibilities are considered. PMID- 12782210 TI - Water maze performance and changes in serum corticosterone levels in zinc deprived and pair-fed rats. AB - The aims of the present study were (1) to evaluate the learning and short- and long-term memory of zinc-deprived (ZD) and pair-fed (PF) rats in a Morris water maze (MWM) and (2) to monitor the serum corticosterone levels of these rats before and after swimming. Young Sprague-Dawley rats (aged 27-31 days) consumed AIN-93G diet for 10 days, and then were separated into ad libitum control (CT), PF and ZD groups. The zinc content of the diet was 25-30 ppm (CT and PF) or <1 ppm (ZD). After 17 days on experimental diets, a MWM was used to test spatial cognition. Delayed-matching-to-place (DMP) test results indicate that both zinc deprivation and food restriction had no effect on short-term memory. The PF rats exhibited significantly impaired learning and thigmotaxia (i.e., wall hugging) in the learning test. The PF group also demonstrated less preference for the target zone in the first 15 s of the probing test. When the total 120 s of the probing test was considered, there were no differences in preference for the target zone, but thigmotaxia was greater in the PF than the CT group. The only behavioral change of the ZD group was thigmotaxia observed during the 120-s probing test following training, indicating the increment of anxiety. Morning basal corticosterone levels before swim training were significantly elevated in the PF group on Day 15 of dietary treatment, whereas a significant elevation of the basal corticosterone level in the ZD group was not statistically significant until Day 22. The data indicate an association between impaired learning, poor searching strategy and elevated corticosterone in the PF group. In contrast, the ZD rats showed normal cognitive performance but had elevated corticosterone and increased anxiety-like behavior (thigmotaxia). PMID- 12782211 TI - Role of gustation in the recognition of oleate and triolein in anosmic rats. AB - Recent studies suggest a chemical perception of dietary fat in the oral cavity. To examine the role of gustation for the recognition of oleate and triolein, very short-term (5-min), two-bottle preference tests were conducted in anosmic rats. To minimize the effects of olfaction, texture and postingestive effects, rats were rendered anosmic with intranasal zinc sulfate, test substances were suspended in 0.3% xanthan gum solution and test fluids were offered for 5 min. Rats preferred oleate fluid but not triolein fluid to the control of 0.3% xanthan gum solution. The preference threshold for oleate in the rat oral cavity was between 0.2% and 0.5%. In the two-bottle preference tests between oleate and triolein, rats preferred oleate fluid to triolein fluid, showing discrimination of oleate and triolein. The results suggest that rat recognizes oleate by a gustatory cue and that fatty acid but not triglyceride is important for gustatory recognition of fat. PMID- 12782212 TI - Differential anxiolytic effect of enalapril and losartan in normotensive and renal hypertensive rats. AB - The effect of angiotensin-converting enzyme (ACE) inhibitor enalapril (EPL) (2 and 4 mg/kg), angiotensin (AT) II receptor antagonist losartan (LRN) (5 and 10 mg/kg), and anxiolytic drug diazepam (DZP) (0.5 mg/kg) on anxiety parameters were evaluated in experimentally induced renal hypertensive rats (RHR). Renal hypertension was induced in Wistar strain male albino rats weighing 200-250 g by following the method of Goldblatt. The animals having systolic blood pressure more than 180-210 mm Hg were subjected to open-field exploratory behaviour, elevated plus maze behaviour, and social interaction tests of anxiety. The RHR showed hyperactivity in open-field behaviour and anxiogenicity in elevated plus maze and social interaction tests. Losartan (5 and 10 mg/kg) and DZP (0.5 mg/kg) significantly attenuated the hyperactivity and anxiogenic behaviour in experimentally induced hypertensive rats and induced anxiolysis in normotensive rats (NTR). Enalapril reversed the hypertension-induced alteration only at higher dose (4 mg/kg) and failed to show any effect in NTR. It can be concluded that renin angiotensin aldosterone system (RAAS) has a significant role on behaviour, and LRN has shown better effect in reversing the hyperactivity and anxiogenicity in the experimentally induced hypertensive rats, indicating a possible role of AT receptor in the mediation of anxiolysis. PMID- 12782213 TI - Sensory-specific satiety is affected more by volume than by energy content of a liquid food. AB - As a food is consumed, its perceived pleasantness declines compared to that of other foods. This phenomenon, referred to as sensory-specific satiety, contributes to the termination of eating, along with other factors. This study tested whether the change in ratings of pleasantness after consuming a food is related to either the amount of food that is consumed or to its energy content. On each of 3 days, 36 women consumed a different formulation of a milk-based liquid food: (1) 300 ml, 2067 kJ; (2) 600 ml, 2067 kJ; or (3) 600 ml, 4134 kJ. The three formulations of the liquid food varied in volume and energy, but were matched for palatability and macronutrient composition. Participants rated the pleasantness of samples of the liquid food and four other foods both immediately before and after consumption of the liquid food. Results showed that doubling the volume of the liquid food that was consumed, without changing the energy content, significantly decreased pleasantness ratings of the liquid food and increased sensory-specific satiety. Doubling the energy content of the food without changing its volume, however, had no additional effect on the decrease in the ratings or on sensory-specific satiety. These results suggest that the volume of food that is consumed has a greater influence on perceptions of a food's pleasantness than does its energy content. Thus, the volume of food may affect the termination of eating in part through effects on sensory-specific satiety. PMID- 12782214 TI - Postoperative haptic training facilitates the retrieval of visual-based memories after visual cortex lesions in rats. AB - Two experiments examined the effects of postoperative haptic discrimination training on the relearning of a maze visual discrimination in rats with visual cortex lesions. In the first experiment, rats learned a visual intensity discrimination prior to ablation of the lateral Oc2L cortex. Lesion rats were exposed to either a rough/smooth haptic discrimination training condition, a random training condition, or a no-training condition prior to relearning the visual task. Lesion rats relearned the visual task faster after haptic training than after other postoperative experiences. The second experiment replicated these procedures but with rats in which most of the visual cortex was removed. The lesion-induced relearning deficits in the second experiment were similar to the deficits seen for the smaller Oc2L lesions in the first experiment, supporting the hypothesis that the lateral visual cortex is critical for intensity discrimination. Haptic training also reduced these deficits, but the magnitude of this effect was related to the characteristics of the haptic cue. Postoperative training with haptic cues can produce specific and nonspecific information transfer from the intact somatosensory system to the damaged visual system that can facilitate the visual relearning. Possible implications for neuropsychological rehabilitation are also discussed. PMID- 12782215 TI - The interaction of bombesin and corticotropin-releasing hormone on ingestive behavior in the domestic fowl. AB - An experiment was conducted to investigate if bombesin (BM) elicits its effects on feeding through the release of corticotropin-releasing factor (CRF). Single Comb White Leghorn (SCWL) and broiler cockerels were stereotaxically implanted with a 23-gauge stainless steel cannula in the right lateral ventricle. Birds were infused with 0 or 0.5 microg BM, 5 microg alphaCRF (a CRF antagonist), or a combination of both. Food and water consumption were monitored at 15-min intervals through 3 h postinjection. Food and water consumption were both significantly decreased by BM in both SCWL and broilers. In SCWL, alphaCRF had no effect on food intake by itself, but attenuated the effects of BM. In broilers, alphaCRF caused a slight, but significant, decrease in food intake, but also attenuated the effects of BM. Water consumption was not affected by alphaCRF in either broilers or SCWL. We conclude that BM may mediate its central effects on food intake in chickens through the release of CRF. PMID- 12782216 TI - Cardiovascular and metabolic responses to fasting and thermoneutrality in Ay mice. AB - Several lines of evidence support a role for reduced melanocortin signaling in the regulation of metabolic rate and cardiovascular function during negative energy balance. We tested the hypothesis that agouti yellow (B6.Cg-A(y)) mice would exhibit blunted physiologic responses to fasting and thermoneutrality. Male B6.Cg-A(y) mice (A(y); n=11, 34+/-2 g) and lean B6 littermates (B6; n=7, 26+/-2 g) were implanted with telemetry devices and housed in metabolic chambers (T(a)=23 degrees C) to determine the effects of a 24-h fasting and exposure to thermoneutrality (T(a)=30 degrees C) on mean arterial pressure (MAP), heart rate (HR), AP and HR variability (time and frequency domain), oxygen consumption (VO(2)), and locomotor activity. A(y) mice exhibited elevated baseline light period MAP (A(y): 113+/-4; B6: 99+/-3 mm Hg) and VO(2) (A(y): 1.82+/-0.08 vs. B6: 1.45+/-0.13 ml/min) with no difference in HR (A(y): 530+/-12 vs. B6: 548+/-19 bpm). At 12-24 h after food removal, A(y) mice displayed normal fasting-induced bradycardia (A(y): -106+/-12; B6: -117+/-19 bpm) and reduction in VO(2) (A(y): 0.19+/-0.04 vs. B6: -0.28+/-0.05 ml/min), but with augmented hypotension (A(y): 9+/-2 vs. B6: -0.5+/-2 mm Hg) and blunted hyperactivity (A(y): 27+/-23 vs. B6: 122+/-42 m/11 h). Fasting was associated with increased HR variability in both time and frequency domain in B6 but not A(y) mice. Exposure to thermoneutrality produced comparable reductions in MAP, HR, and VO(2) in both strains. We conclude that inhibition of melanocortin signaling is not requisite for, but participates in, the metabolic and cardiovascular responses to negative energy balance. PMID- 12782217 TI - A paper screening test to assess genetic taste sensitivity to 6-n propylthiouracil. AB - The purpose of this study was to develop a quantifiable method to fix 6-n propylthiouracil (PROP) onto filter paper disks and to test the validity of the method relative to the three-solution test, previously developed in this laboratory. Filter paper disks were impregnated with 50 mmol/l PROP or 1.0 mol/l NaCl then dried. The concentration of PROP per disk was determined to be 0.280 mg+/-2.2% (CV) by ethanol extraction. Subjects were studied in two groups (n=62 per group). All subjects were initially classified by the three-solution test and then were independently classified by the paper disk test. In the paper disk test, group means for PROP (+/-95% confidence interval) were calculated and used to establish numerical cutoff scores for taster group classification. Cutoff scores for Group 1 were used to classify subjects in Group 2, verifying that the cutoffs were applicable to another subject group. The percentage of nontasters, medium tasters, and supertasters identified by paper disk was 27%, 42%, and 31%, respectively, which is consistent with expected frequencies in the population. For Group 1, the agreement between the two classification methods ranged from 86% to 94% across taster groups. For Group 2, the agreement ranged from 83% to 100%. The contingency coefficient (P) of the degree of association between the two classification methods was high (P=0.77 and P=0.74 for Groups 1 and 2, respectively; P< or =.001). These results demonstrate that the paper disk method is a reliable screening tool for assessing sensitivity to PROP that has numerous applications in basic and applied research. PMID- 12782218 TI - Behavioral effects of static high magnetic fields on unrestrained and restrained mice. AB - High-strength static magnetic fields are common tools in clinical imaging, but the behavioral effects are not well characterized. Previous studies on rats showed that fields of 7 T or above produced locomotor circling, conditioned taste aversion (CTA) and c-Fos in vestibular nuclei. To determine the generality of the behavioral effects on a smaller species, we subjected restrained or unrestrained mice to 30-min exposures in a 14.1-T field. Mice were given saccharin immediately prior to magnet or sham exposure on 3 consecutive days. All mice exposed to the magnet developed a CTA, and a significant number displayed tight circling and suppression of rearing. Unrestrained mice exhibited larger effects than restrained mice. These effects, similar to the effects in rats, may be the result of a vestibular disturbance caused by the magnetic field. PMID- 12782219 TI - Compulsive behavior in the 5-HT2C receptor knockout mouse. AB - The efficacy of serotonergic pharmacotherapy indicates that serotonin (5-HT) plays a role in the treatment, if not the etiology, of obsessive-compulsive disorder (OCD). While some clinical evidence implicates 5-HT(2C) receptors in this disorder, a definitive function has yet to be validated. We hypothesized that 5-HT(2C) receptor knockout (KO) mice may display compulsive-like behavior. This paper describes characterization of several distinct, highly organized behaviors in mice lacking functional 5-HT(2C) receptors, which supports a compulsive-like syndrome.Compulsive-like behavior was assessed in male 5-HT(2C) receptor KO and wildtype (WT) mice. Chewing of non-nutritive clay, chewing patterns on plastic-mesh screens, and the frequency of head dipping were measured. 5-HT(2C) receptor KO mice chewed more clay, produced a distinct pattern of "neat" chewing of plastic screens and exhibited reduced habituation of head dipping activity compared to WT mice. We conclude that the 5-HT(2C) receptor null mutant mouse provides a promising model of compulsive behavior and a means to further explore the role of 5-HT in OCD. PMID- 12782220 TI - LPS fever in old rats depends on the ambient temperature. AB - In earlier work, we found that following lipopolysaccharide (LPS) injection at an ambient temperature (Ta) of 23 degrees C, old rats developed blunted fevers compared with those of young rats. However, the old rats did become febrile if placed in a thermally graded alleyway: they spent more time in the warm end of the gradient and developed a significantly higher body temperature (Tb) than they did following saline injections. In the present experiments, we maintained old and young rats for 3 days at 20 or 31 degrees C (the Ta preferred by the old rats given LPS). After LPS (50 microg/kg ip), the young rats developed equivalent fevers at both Ta's. The old rats developed fevers that were equivalent to those of the young rats at 31 degrees C. At 20 degrees C, their fever was significantly lower. These results suggest that Ta plays a decisive role in the ability of old rats to mount febrile responses. PMID- 12782221 TI - Fasting-induced increases in food hoarding are dependent on the foraging-effort level. AB - Two strategies that have evolved to help animals meet energy demands are increases in body fat and in hoarded food. Reliance on each varies, but both are characterized by energy stored in excess of current demands for future use. Fasted Siberian hamsters decrease their lipid stores and, upon refeeding, food hoarding rather than food intake increases. Here, we tested the effect of foraging-effort level on fasting-induced increases in food hoarding and whether decreases in total body fat or individual fat pad masses were associated with the hoarding increases. This was accomplished by housing female Siberian hamsters in a foraging/hoarding system where they earned food pellets upon completion of a programmed number of wheel revolutions (10, 75 or 200 per 75-mg pellet), or had no foraging requirement (free food and an active or blocked running wheel). After baseline, half the hamsters in each group were food deprived for 32 h and then refed (control hamsters were killed after the fast without refeeding). Fasted refed hamsters increased foraging and food hoarding, especially at the lowest foraging effort, but not food intake, with few exceptions. These responses became progressively smaller as foraging effort increased. Fasting induced similar losses in carcass lipid across foraging efforts, but the lipid loss was not uniform across the fat pads and was not clearly related to hoard size. Collectively, the fasting-induced, increased food hoarding may be triggered by an overall change in energy metabolism rather than by a general or fat-pad-specific decrease in lipid stores. PMID- 12782222 TI - Simmondsin: effects on meal patterns and choice behavior in rats. AB - Simmondsin, a glycoside from jojoba meal, decreases food intake after oral administration. The present experiments are designed to clarify the mechanism of simmondsin's anorectic activity. The meal pattern analysis shows that simmondsin supplementation at different doses results in a dose-dependent food intake reduction, which is more pronounced after prior simmondsin experience. The effect of simmondsin on meal patterns (decreased meal size, meal duration and eating rate, increased latency to eat) is most severe at the highest concentration. Rats familiar with simmondsin more seriously postpone their first meal than with first contact, resulting in a decrease of the meal frequency and the day/night feeding ratio. Rats given the choice between a control diet and a simmondsin-supplemented (0.5%) diet, after half an hour, have a significant preference for the control diet. Simmondsin seems to have a specific flavor when mixed in the food since rats recognise the feeder containing simmondsin. The ability of simmondsin to induce conditioned taste aversion (CTA) was also investigated. Rats receiving simmondsin at concentrations of 0.15%, 0.25% or 0.5% during their conditioning develop significant taste aversions to the saccharin solutions. The performed experiments indicate that the simmondsin activity shows some analogy with the satiating molecule cholecystokinin (CCK) at first contact, but shows more analogy with the illness-inducing agent lithium chloride (LiCl) after prior experience with simmondsin. Rats familiar with simmondsin avoid simmondsin-supplemented food by directly monitoring its presence, and by learning to relate it to the postingestive consequences of consumption. PMID- 12782223 TI - Lipopolysaccharide induces biochemical alterations in chicks trained on the passive avoidance learning task. AB - We have recently shown that activation of the immune system with lipopolysaccharide (LPS) results in memory-processing deficits for the passive avoidance learning task in the day-old chick. The current study examined two important issues in understanding the mechanisms underlying these memory deficits associated with immune system activation, namely, whether LPS (1) impairs Na(+)/K(+)-ATPase functioning and (2) increases corticosterone (CORT) concentrations in chicks trained on the task. As the effects of LPS on sickness behavior have only previously been characterized in older chickens, this study also tested whether LPS is able to produce similar alterations in day-old chicks. LPS decreased brain Na(+)/K(+)-ATPase activity and increased plasma concentrations of CORT in chicks trained on the passive avoidance learning task. These findings give an insight into some of the mechanisms that may be responsible for the LPS-induced memory-processing deficits. Consistent with previous research in older chickens, LPS increased body temperature in a dose dependent manner, however, only the lowest dose of LPS tested significantly decreased food intake in the day-old chicks. PMID- 12782224 TI - Salt intake by normotensive and spontaneously hypertensive rats: two-bottle and lick rate analyses. AB - Spontaneously hypertensive rats (SHR) overconsume NaCl compared to the normotensive Wistar Kyoto rat (WKY) rat. In the present experiment, two-bottle preference for NaCl (0.01, 0.03, 0.1, 0.3, 0.5, 1.0, 3.0 M) and lick rate analyses were used to identify the possible mechanisms that underlie the intake of NaCl by male SHR. Two-bottle preference and absolute NaCl intake by SHR were greater than that of WKY rats. When NaCl intake was calculated on the basis of body weight, SHR consumed more NaCl per 100 g body weight than did WKY. Also, during the one-bottle test, SHR consumed more 0.1 and 0.3 M NaCl per 100 g body weight than did WKY. The increased intake of NaCl by SHR was most evident for 0.3 M NaCl. Intake is determined by the initial rate of licking and the decline in lick rate over time. Nonlinear regression analysis of lick rate showed that the initial lick rates (licks/min) were similar for male WKY and SHR. Lick rate declined more rapidly when WKY rats drank 0.3 M than when they drank 0.1 M NaCl, a result consistent with the role of negative feedback in controlling the decay in lick rate. This concentration-dependent change in lick rate was not seen in SHR. Also, SHR lick rate for 0.1 and 0.3 M NaCl decelerated more slowly than that of WKY rats. The increased intake of hypertonic NaCl by SHR was due to a decrease in the decline in lick rate, suggesting that SHR are less responsive to ingestion contingent negative feedback. PMID- 12782225 TI - Anogenital distance measured at weaning is correlated with measures of blood chemistry and behaviors in 450-day-old female mice. AB - In female mice, anogenital distance (AGD), measured at weaning, provides an estimate of uterine exposure to testosterone from flanking male mouse littermates. A variant of the anogenital distance index (AGDI) that uses the residual value of AGD after accounting for the effect of weight by regression (AGDWTRES) was measured at weaning in F(2) female mice from a C57BL/6J x DBA2/J cross. AGDWTRES was used to examine the relationship between intrauterine environment and blood chemistry variables and activity-related behaviors when the females were 450 days old. Longer AGDWTRES values correlated with lower levels of calcium, cholesterol, phosphorus, iron, and protein, which is opposite to the expected direction, based on underlying sex differences for blood chemistry. A positive correlation was found between AGDWTRES and two activity-related measures (the number of rears in a test of exploration, and the number of sectors of a rod that are entered by the mouse). These findings suggest that in utero proximity to males, as indexed by AGDWTRES, may have effects on fundamental aspects of blood chemistry and behavior that extend well into mouse middle age, and could play an important role in health. PMID- 12782227 TI - Forebrain sites of NPY action on estrous behavior in Syrian hamsters. AB - Food deprivation and similar metabolic challenges inhibit estrous behavior in female Syrian hamsters. The relevant metabolic cues appear to be detected in the hindbrain, and this information is then relayed synaptically to the forebrain circuits controlling estrous behavior. Neuropeptide Y (NPY) may be one of the neuropeptides/neurotransmitters serving this function. Infusion of NPY or the Y2/Y5 agonist, peptide YY3-36 (PYY3-36), into the lateral ventricles rapidly inhibits estrous behavior in ovariectomized, steroid-primed hamsters. This experiment sought to determine the neural loci where NPY acts to inhibit estrous behavior. Steroid-primed animals received infusions of artificial cerebrospinal fluid (aCSF) vehicle, 0.024 nmol PYY3-36 and 0.24 nmol PYY3-36 in separate tests 30 min prior to testing for sexual receptivity. Infusion of 0.24 nmol, but not 0.024 nmol, of PYY3-36 reduced lordosis duration when infused into the paraventricular nucleus of the hypothalamus (PVN), the caudal part of the medial preoptic area (MPO), the anterior hypothalamus (AH) or the lateral ventricles. Placements in the ventromedial hypothalamus (VMH), the arcuate nucleus (ARC) and the fourth ventricle were generally without effect. These data suggest that increased endogenous release of NPY into the caudal MPO-AH-PVN continuum during food deprivation could contribute to the observed inhibition of sexual receptivity. The possible contributions of other neuropeptides and neural estrogen receptors to this action of NPY are discussed. PMID- 12782226 TI - Behavioral and cardiovascular changes in the chronic mild stress model of depression. AB - Depression is a multifaceted psychological disorder that involves changes in behavior, neuroendocrine function, and physiological responses. The present study investigated multiple behavioral and cardiovascular consequences in the chronic mild stress (CMS) rodent model of depression. Rats were exposed to 4 weeks of CMS followed by 4 weeks of a stress-free recovery period. Sucrose intake, a measure of anhedonia, and spontaneous locomotor activity were measured weekly throughout the study, and cardiovascular function tests were conducted at the completion of the protocol. The results indicate that CMS results in anhedonia and reduced locomotor activity, as well as elevated heart rate (HR), reduced HR variability, and elevated sympathetic cardiac tone. The behavioral effects of CMS recovered to baseline (prestress) levels during the recovery period; however, cardiovascular changes were observed following the recovery of sucrose intake and activity levels. The present findings suggest that behavioral changes that are indicative of anhedonia and locomotor alterations associated with depression are dissociable from long-term cardiovascular changes induced by CMS. PMID- 12782228 TI - Social regulation of reproduction in the female mound-builder mouse (Mus spicilegus). AB - Social environment influences the reproductive physiology and sexual behaviour of the female house mouse Mus musculus. An all-female environment tends to suppress the oestrous cycles, whereas the presence of a male induces and synchronises sexual receptivity. However, reproductive responses to social environment may differ among the various species of rodents. In mound-builder female mice, Mus spicilegus, periods of sexual receptivity are interrupted by periods where adult females display a vaginal closure. We investigated the influence of different social environments on the vaginal opening and oestrous state of adult female M. spicilegus. Result showed that when females were grouped their vaginas were generally closed but that vaginal opening occurred when they were isolated or housed with a sexual partner. Females became sexually receptive when housed with a male, but when isolated their cervical smears did not reach characteristics of the oestrus. In female M. spicilegus, male presence thus has a stimulating effect on oestrous induction. Furthermore, cohabitation with females has an inhibiting effect on vaginal opening. PMID- 12782229 TI - Fearfulness and sex in F2 Roman rats: males display more fear though both sexes share the same fearfulness traits. AB - The pattern of sex differences in a large sample (about 400 for each sex) of F2 generation rats, derived from inbred Roman high- and low-avoidance strains differing in fearfulness and brain functioning, was investigated. We obtained measures from responses to a battery of novel/threatening tests [open field (OF), plus maze (PM), hole board (HB), activity (A), and acoustic startle reflex (ASR)] as well as learned fear paradigms [classical fear conditioning (CFC) and shuttlebox avoidance conditioning (SAC)]. The results showed that almost all behaviors assessed fit with a pattern of unidirectional sex effects characterized by male rats as being more fearful than females: males defecated more than females in the OF, PM, HB, ASR, and CFC; ambulated less in the OF, PM, A, and SAC; showed more self-grooming in PM and HB; explored the open arms of the PM and the holes of the HB less; displayed enhanced ASR; and showed poorer performance in the SAC task. We applied two factor analyses to each sex showing that, in general, they shared a common three-factor structure: a Learned Fear Factor comprising SAC and CFC responding, a Fear of Heights/Open Spaces Factor with the highest loadings for open arm behavior in the PM, and an Emotional Reactivity Factor, mainly grouping defecations, ambulation, and self-grooming. These results indicate that the essential components of fearful behavior are similar for both sexes in an inbred but genetically heterogeneous population. PMID- 12782230 TI - c-Fos expression in supramammillary and medial mammillary nuclei following spatial reference and working memory tasks. AB - To investigate brain substrates of spatial memory, neuronal expression of c-Fos protein was studied. Two groups of rats were trained in two spatial memory tasks in the Morris water maze, where the rats have to apply a reference memory rule or a working memory rule. In addition to the experimental groups, two control groups were used to study c-fos activation not specific to the memory processes studied. After immunohistochemical procedures, the number of c-Fos positive neuronal nuclei was quantified in the mammillary body (MB) region (medial mammillary nucleus [MMn] and supramammillary nucleus [SuM]). The results have shown that some MMn neurons expressed c-Fos nuclear immunoreactivity related to spatial working memory but not to spatial reference memory. The increased number of c-Fos immunoreactive neuronal nuclei in the SuM was related to spatial training but not to either working or reference memory demands of the tasks. PMID- 12782231 TI - Electromyographic activity of the lip muscle as a measure of puffing on a cigarette. AB - The electromyographic activity (EMG) of the upper lip muscle (m. orbicularis oris) during sucking on a cigarette was examined as a potential new measure of smoking behaviour. This parameter was examined under conditions of actual smoking, sham smoking and sucking on a straw. We applied psychophysical procedures (methods of magnitude production), normal and paced smoking, two conventional means to confirm puffing (video recording and changes in air pressure in a cigarette holder) on different groups of smokers (heavy and light). The data revealed that lip-EMG is a valid and reliable means to quantify sucking on a cigarette in a variety of situations common to the study of smoking. It was concluded that lip-EMG may be a sensitive and selective measure of motor activity associated with smoking. The lip-EMG may also be applied together with hand activity and breathing to identify individual puffing, particularly in heavy smokers and during actual smoking. Discussed were several applications of this method to the study of smoking and other motivated behaviour such as consumption of water and palatable fluids. PMID- 12782232 TI - A dopamine D2 receptor gene polymorphism and physical activity in two family studies. AB - A role for dopamine neurotransmission in the regulation of motor activity and reinforcement of behavior is supported by considerable evidence. We studied the association between a marker in the dopamine D2 receptor gene (DRD2) and physical activity level in two cohorts. A first cohort consisted of 721 participants from 161 families of the Quebec Family Study (QFS). Physical activity phenotypes were obtained from a three-day diary and a questionnaire probing physical activity during the past year. The second cohort was the HERITAGE Family Study (HERITAGE), which included 275 Black and 497 White participants from 228 families, among whom past year leisure time and occupational physical activity were probed. A fragment length polymorphism in exon 6 of the DRD2 gene was detected by the polymerase chain reaction (PCR) and NcoI digestion. Frequencies for the T and C alleles were 28% and 72% in the QFS. In the QFS, TT homozygote women had 25% and 34% lower age and BMI-adjusted physical activity level during the past year, compared to CC homozygotes and CT heterozygotes (F=4.42, P=.016). The DRD2 genotype was not associated with the QFS phenotypes obtained from the three-day diary. In the HERITAGE, the frequency of the T allele was 30% among Whites and 63% among Blacks. Similarly, the TT homozygote White women had 29-38% lower sports index (F=4.09, P=.023) and 27-33% lower work index (F=6.23, P=.004) than the CC homozygotes and CT heterozygotes. The results suggest that DNA sequence variation in the DRD2 gene is associated with physical activity levels among White women. PMID- 12782233 TI - Dietary fish oil does not protect rats exposed to restraint or sleep deprivation stress. AB - It has been suggested that fish oil (FO) prevents weight loss caused by physiological stress such as cancer, injury, or cardiovascular disorders. Previously, we observed that a high-fat diet containing corn and coconut oil exaggerated weight loss caused by the mixed physiological and psychological stress of repeated restraint (RR). This experiment tested the effects of a high fat diet containing FO as the predominant lipid source in rats exposed to the mixed physiological and psychological stress of either RR or sleep deprivation (SD). FO did not prevent stress-induced hypophagia or weight loss in RR or SD rats but exaggerated the negative effects of stress on body weight in SD rats by promoting loss of lean body mass. RR caused a reduction in body fat content irrespective of dietary treatment. In SD rats, both stress and FO independently reduced body fat mass. FO did not have any effect on adrenal and thymus weights during RR or SD and did not influence corticosterone levels after 1 h of RR or after 48 or 96 h of SD. In conclusion, our results suggest that high levels of dietary FO do not improve the response to stress in rats exposed to mixed stressors. PMID- 12782234 TI - Fasting plasma triglyceride levels and fat oxidation predict dietary obesity in rats. AB - We investigated whether fuel metabolism prior to high-fat feeding differs in outbred Sprague-Dawley rats either prone or resistant to diet-induced obesity. Chow-fed rats were deprived of food, and blood was collected 12, 18, and 24 h later. Rats were then fed a high-fat diet ad libitum for up to 4 weeks to assess weight gain. Blood samples were analyzed for a variety of metabolic fuels and hormones. Only fasting plasma triglyceride concentrations showed a positive correlation with the weight gain during the high-fat feeding period, with concentrations after 18 h of fasting showing the most consistent relationship to weight gain. Body weights and fat pad weights did not correlate with fasting plasma triglyceride concentrations before high-fat feeding. The amount of 14CO(2) recovered from gavaged [14C]palmitic acid in chow-fed rats negatively correlated with weight gain during the subsequent period of high-fat feeding. These results show that there are preexisting differences in fat catabolism that may underlie differential susceptibility to diet-induced obesity; in particular, fasting levels of plasma triglycerides and fatty acid oxidation may be early predictive markers for this susceptibility. PMID- 12782235 TI - Parameters for quantification of hunger in broiler breeders. AB - The commercial restricted feeding programme of broiler breeders has a major negative effect on welfare, as the birds are continuously hungry. Objective parameters of hunger are needed to evaluate new management or feeding systems that may alleviate hunger and thus improve broiler breeder welfare. The aim of this experiment was to develop such parameters. Female broiler breeders (Hybro G), n=10 per treatment, were subjected to different levels of feed restriction and we assumed that we thus induced different levels of hunger. Treatment groups were fed ad libitum and at 90%, 70%, 50%, 35% and 25% of ad libitum. In addition, female broiler breeders that were less intensively selected on growth and food conversion (JA57, n=10 per treatment) were fed ad libitum and at 70% of ad libitum. At 6-7 weeks of age, home pen behaviour, behaviour in the open field and baseline plasma corticosterone, glucose and nonesterified fatty acid (NEFA) concentrations were determined. Thereafter, birds were subjected to the feed intake motivation (FIM) test that measures compensatory feed intake. Linear relationships between the level of restriction and the compensatory feed intake on the first days of the FIM test and the glucose/NEFA ratio were found which indicate that these parameters can be used to quantify the level of hunger. Curvilinear relationships between sitting and standing in the home pen, walking in the open field and level of restriction were found. A third-grade hyperbolic relationship between the level of restriction and the baseline plasma corticosterone concentration was found. Thus, these parameters do not seem to represent a straightforward relation with the level of hunger. Intensive selection on growth and food conversion seems to have affected the behaviour but not the physiological responses to feed restriction, because the JA57 birds responded behaviourally, but not physiologically, different to feed restriction as compared with the Hybro birds. PMID- 12782236 TI - Task solving by procedural strategies in the Morris water maze. AB - The aim of the present work was to assess the importance of the "general procedural" components, when for rats it was impossible to employ extramaze allothetic information to reach the goal in the Morris water maze (MWM). Groups of Long-Evans rats (males, 70 days old) were trained (10 trials per day, over five consecutive days) following seven paradigms. Four paradigms differed in context (extramaze cues available; extramaze cues not available) and in platform location (constantly at the center of one quadrant of the water maze; at random at the center of any one of the quadrants). In the fifth paradigm, there were no extramaze cues available, and the platform was located at random distances from the maze wall. In the sixth paradigm, rats underwent the standard MWM training (extramaze cues available, invisible platform constantly placed in the center of one quadrant) but they were administered with scopolamine before the daily trials. In a seventh paradigm, the platform was visible. In all paradigms, the starting point was randomized with respect to the goal. When platform distance from the wall was random, there was no significative better performance after the trials. In all the six paradigms in which platform location was at a constant distance from the wall the times spent before reaching the platform decreased progressively, to become constant on Days 4 and 5. The groups which could not employ the allothetic extramaze component (extramaze cues not available; changing of the quadrant of platform location; scopolamine administration) showed a progressively better performance even though their delays on the last 2 days were longer than those of the "standard MWM" and "visible platform" groups. The slightly less efficient performance is attributable to the rat's search strategy, a "subcircular" swimming pattern within the geometric limits of the central areas of the quadrants, where the platform was constantly placed. That no extramaze allothetic information was employed is shown by the finding that on Day 6 (probe test: 90 s in the tank without platform) no animals exhibited preference for any quadrant, while the "standard MWM" group did show such a preference. It can be concluded that rats under conditions of constant relationship of the goal to the contours of the pool employ search strategies based on general procedural components. PMID- 12782237 TI - Macronutrient selection through postingestive signals in sea bass fed on gelatine capsules. AB - The role of the food orosensory properties on protein (P), fat (F) and carbohydrate (CH) self-selection was investigated in fish fed gelatine capsules containing pure macronutrients. A total of 40 sea bass (39.6+/-6.2 g initial body weight) distributed in eight 75-l tanks were used. In a sequence of experimental phases, sea bass were fed a pelleted complete diet, an encapsulated complete diet or a combination of separately encapsulated pure macronutrients. In order to induce associative learning, capsules containing a given macronutrient were paired with a particular colour. Our results demonstrate that fish are able to regulate food intake, so as to balance their energy intake, when they are fed a complete encapsulated diet and, therefore, without using the orosensory properties of the diet. Moreover, sea bass learn to discriminate and select among colour-coded, pure macronutrient capsules to compose a complete and balanced diet using colour as the only external cue. The composition of selected diet was 55% P, 23% CH and 22% F in terms of macronutrient percentage. The diet orosensory properties do not seem to be necessary to regulate macronutrient intake either, suggesting that an associative learning between capsule colour and content can be established through monitoring of macronutrient intake by postingestive and/or postabsorptive mechanisms. These results provide the first insight into energy and macronutrient self-selection by fish fed on gelatine capsules containing separate macronutrients. PMID- 12782238 TI - A comparison of the effects of added saliva, alpha-amylase and water on texture perception in semisolids. AB - The effect of adding saliva or a saliva-related fluid (alpha-amylase solution and water) to custard prior to ingestion on the sensory ratings of odour, flavour and lip-tooth-, mouth- and after-feel sensations was investigated. Saliva had previously been collected from the subjects and each subject received his/her own saliva. Sixteen subjects from a trained panel assessed 17 flavour and texture attributes of soy- and milk-based custard desserts. Immediately prior to administration, two different volumes (0.25 and 0.5 ml) of three different saliva related fluids (saliva, alpha-amylase solution and water) were added to the product. The added volumes represented an approximately 33% and 66% increase of the volume of saliva present in the mouth during ingestion. The results show that addition of a fluid affected the mouth-feel attributes of melting, thickness and creamy. Melting was the only attribute on which the type of fluid had an effect, where saliva elicited a stronger melting effect than the alpha-amylase solution and water. The volume of the added fluid affected a number of attributes (thick and creamy mouth-feel and fatty after-feel). It can be concluded that in general the sensory attributes of semisolids were relatively stable. Mouth- and after feel sensations were partly affected, while odour, flavour and lip-tooth-feel sensations were not affected by an increase in volume of saliva or other saliva related fluid during ingestion. PMID- 12782239 TI - The surrogate nipple technique in the rat provides a useful animal model of suckling in bottle-feeding circumstances: reply to Blass (2002). PMID- 12782241 TI - The human cytomegalovirus. AB - Human cytomegalovirus (HCMV), a betaherpesvirus, represents the major infectious cause of birth defects, as well as an important pathogen for immunocompromised individuals. The viral nucleocapsid containing a linear double-stranded DNA of 230 kb is surrounded by a proteinaceous tegument, which is itself enclosed by a loosely applied lipid bilayer. Expression of the HCMV genome is controlled by a cascade of transcriptional events that leads to the synthesis of three categories of viral proteins designated as immediate-early, early, and late. Clinical manifestations can be seen following primary infection, reinfection, or reactivation. About 10% of infants are infected by the age of 6 months following transmission from their mothers via the placenta, during delivery, or by breastfeeding. HCMV is a significant post-allograft pathogen and contributes to graft loss independently from graft rejection. Histopathologic examination of necropsy tissues demonstrates that the virus enters via the epithelium of the upper alimentary, respiratory, or genitourinary tracts. Hematogenous spreading is typically followed by infection of ductal epithelial cells. Infections are kept under control by the immune system. However, total HCMV clearance is rarely achieved, and the viral genome remains at selected sites in a latent state. Virological and molecular detection of HCMV, as well as serological demonstration of a specific immune response, are used for diagnosis. Treatment of HCMV infections is difficult because there are few options. The presently available drugs produced a significant clinical improvement, but suffer from poor oral bioavailability, low potency, development of resistance in clinical practice, and dose-limiting toxicities. PMID- 12782242 TI - Formation and plasticity of GABAergic synapses: physiological mechanisms and pathophysiological implications. AB - gamma-Aminobutyric acid(A) (GABA(A)) receptors mediate most of the fast inhibitory neurotransmission in the CNS. They represent a major site of action for clinically relevant drugs, such as benzodiazepines and ethanol, and endogenous modulators, including neuroactive steroids. Alterations in GABA(A) receptor expression and function are thought to contribute to prevalent neurological and psychiatric diseases. Molecular cloning and immunochemical characterization of GABA(A) receptor subunits revealed a multiplicity of receptor subtypes with specific functional and pharmacological properties. A major tenet of these studies is that GABA(A) receptor heterogeneity represents a key factor for fine-tuning of inhibitory transmission under physiological and pathophysiological conditions. The aim of this review is to highlight recent findings on the regulation of GABA(A) receptor expression and function, focusing on the mechanisms of sorting, targeting, and synaptic clustering of GABA(A) receptor subtypes and their associated proteins, on trafficking of cell-surface receptors as a means of regulating synaptic (and extrasynaptic) transmission on a short-time basis, on the role of endogenous neurosteroids for GABA(A) receptor plasticity, and on alterations of GABA(A) receptor expression and localization in major neurological disorders. Altogether, the findings presented in this review underscore the necessity of considering GABA(A) receptor-mediated neurotransmission as a dynamic and highly flexible process controlled by multiple mechanisms operating at the molecular, cellular, and systemic level. Furthermore, the selected topics highlight the relevance of concepts derived from experimental studies for understanding GABA(A) receptor alterations in disease states and for designing improved therapeutic strategies based on subtype-selective drugs. PMID- 12782243 TI - Evolution, structure, and activation mechanism of family 3/C G-protein-coupled receptors. AB - G-protein-coupled receptors (GPCRs) represent one of the largest gene families in the animal genome. These receptors can be classified into several groups based on the sequence similarity of their common heptahelical domain. The family 3 (or C) GPCRs are receptors for the main neurotransmitters glutamate and gamma aminobutyric acid, for Ca(2+), for sweet and amino acid taste compounds, and for some pheromone molecules, as well as for odorants in fish. Although none of these family 3 receptors have been found in plants, members have been identified in ancient organisms, such as slime molds (Dictyostelium) and sponges. Like any other GPCRs, family 3 receptors possess a transmembrane heptahelical domain responsible for G-protein activation. However, most of these identified receptors also possess a large extracellular domain that is responsible for ligand recognition, is structurally similar to bacterial periplasmic proteins involved in the transport of small molecules, and is called a Venus Flytrap module. The recent resolution of the structure of this binding domain in one of these receptors, the metabotropic glutamate 1 receptor, together with the recent demonstration that these receptors are dimers, revealed a unique mechanism of activation for these GPCRs. Such data open new possibilities in the development of drugs aimed at modulating these receptors, and raise a number of interesting questions on the activation mechanism of the other GPCRs. PMID- 12782244 TI - Prevention of sudden cardiac death by n-3 polyunsaturated fatty acids. AB - This is a review of our present understanding of the mechanism by which the n-3 polyunsaturated fatty acids (PUFA) in fish oils prevent fatal ventricular arrhythmias in animals and cultured heart cells. A brief review of three clinical trials that suggest that these PUFAs prevent sudden cardiac death is also included in order to emphasize the potential importance of these fatty acids in human nutrition. The PUFAs act by stabilizing electrically every cardiac myocyte by modulating conductance of ion channels in the sarcolemma, particularly the fast, voltage-dependent sodium current and the L-type calcium currents, though other ion currents are also affected. Work in progress suggests that the primary site of action of the PUFAs may be on the phospholipid bilayer of the heart cells in the microdomains through which the ion channels penetrate the membrane bilayer in juxtaposition with the ion channels rather than directly on the channel protein itself. These PUFAs then allosterically alter the conformation and conductance of the channels. Both potential benefits and possible adverse effects of the PUFAs in man will be discussed. Knowing that the ion channels have been structurally conserved among all excitable tissues, we tested their effects on the electrophysiology of rat hippocampal CA1 neurons and found that the sodium and calcium ion channels in these neurons were also affected by PUFAs. An attempt to show the place of the PUFAs in human nutrition during the 2-4 million years of our evolution will conclude the review. PMID- 12782245 TI - The basic aspects of therapeutics in amyotrophic lateral sclerosis. AB - Once thought to be a single pathological disease state, amyotrophic lateral sclerosis (ALS) is now recognized to be the limited phenotypic expression of a complex, heterogeneous group of biological processes, resulting in an unrelenting loss of motor neurons. On average, individuals affected with the disease live <5 years. In this article, the complex nature of the pathogenesis of ALS, including features of age dependency, environmental associations, and genetics, is reviewed. Once held to be uncommon, it is now clear that ALS is associated with a frontotemporal dementia and that this process may reflect disturbances in the microtubule-associated tau protein metabolism. The motor neuron ultimately succumbs in a state where significant disruptions in neurofilament metabolism, mitochondrial function, and management of oxidative stress exist. The microenvironment of the neuron becomes a complex milieu in which high levels of glutamate provide a source of chronic excitatory neurotoxicity, and the contributions of activated microglial cells lead to further cascades of motor neuron death, perhaps serving to propagate the disease once established. The final process of motor neuron death encompasses many features of apoptosis, but it is clear that this alone cannot account for all features of motor neuron loss and that aspects of a necrosis-apoptosis continuum are at play. Designing pharmacological strategies to mitigate against this process thus becomes an increasingly complex issue, which is reviewed in this article. PMID- 12782246 TI - Antagonism of corticotropin-releasing factor attenuates the enhanced responsiveness to stress observed during protracted ethanol abstinence. AB - One of the most critical attributes of chronic abstinence from alcohol is a state of anxiety, which can lead to mood disturbances and negative affect that can last for months or even years in alcoholics. Within hours after their final exposure to ethanol in experimental conditions, laboratory animals also exhibit an anxiety like state. This state is accompanied by an enhanced stress response and can persist for weeks after withdrawal. One possible mechanism underlying these behavioral changes observed weeks after withdrawal is increased corticotropin releasing factor (CRF) activity. In the present study, we sought to examine the role of CRF in the regulation of behavior in the elevated plus-maze during protracted abstinence by using intracerebroventricular administration of the CRF receptor antagonist [D-Phe(12),Nle(21,38),CalphaMeLeu(37)]rCRF((12-41)) (D-Phe CRF((12-41))). Rats were surgically implanted with a guide cannula aimed at the lateral ventricles and subsequently fed a nutritionally complete ethanol [10% (vol./vol.)] or control liquid diet for 21 days. Rats were further divided into groups receiving microinjections of D-Phe-CRF((12-41)) or vehicle and 15 min of restraint stress, or D-Phe-CRF((12-41)) or vehicle and no restraint. Six weeks after removal of the liquid diet, rats were injected and then placed in a restraint tube or returned to their home cages for 15 min before testing in the elevated plus-maze. Rats with a history of ethanol dependence explored the open arms of the plus-maze significantly less when exposed to restraint stress compared with findings for all other groups, an effect attenuated by pretreatment with D-Phe-CRF((12-41)). Results of the current experiment demonstrated that continuous exposure to ethanol over a 3-week period leads to an increased behavioral responsiveness to stress, which seems to be regulated by CRF. PMID- 12782247 TI - Freely accessible water does not decrease consumption of ethanol liquid diets. AB - In experimental studies, liquid ethanol diets are usually given as the sole source of nutrition and fluid. Two series of experiments were conducted to examine the effect of freely accessible water on the consumption of ethanol liquid diets in male Long-Evans rats. The consumption of diets and subsequent learning ability of rats were first examined in animals given twice-daily saline injections. One group received diet with no access to water for 12 weeks and was subsequently given free access to water with diets for an additional 12 weeks. A second group was given diet and water ad libitum for 24 weeks. Control animals received an isocaloric sucrose-containing diet (with or without ad libitum access to water). Subsequently, rats were tested for active avoidance learning. In the first 12 weeks, animals with ad libitum access to water drank more diet than did water-restricted animals, and previously water-restricted animals increased their diet consumption when access to water was freely available. All water-restricted animals, in both ethanol- and sucrose-treated groups, showed deficits in active avoidance learning, whereas only ethanol-treated animals in groups with ad libitum access to water showed learning deficits. In the second series of experiments, the effect of saline injections on diet consumption, both in the presence and absence of water, was examined. Although saline injections were associated with decreased diet consumption, there was no effect of free access to water. No differences in blood ethanol concentration were seen among groups. Findings obtained from both series of studies demonstrate that consumption of a Sustacal-based liquid ethanol diet does not decrease if access to water is freely available. PMID- 12782249 TI - Effects of chronic ethanol treatment on the angiotensin II-mediated p42/p44 mitogen-activated protein kinase and phosphorylase a activation in rat hepatocytes. AB - We have demonstrated previously that 24 h of ethanol treatment potentiates angiotensin II (ANG II)-stimulated p42/p44 mitogen-activated protein kinase (MAPK) activity in hepatocytes. This potentiation of p42/p44 MAPK by ethanol exhibited agonist selectivity. To compare the effects of acute (24 h) versus chronic (6 weeks) ethanol treatment, ANG II-induced intracellular signaling was examined in (1) rat hepatocytes treated with ethanol for 24 h and (2) hepatocytes obtained from rats fed ethanol for 6 weeks. In hepatocytes obtained from rats fed ethanol for 6 weeks, ANG II-stimulated phosphorylase a was reduced, and this activity was calcium dependent and p42/p44 MAPK independent. Surprisingly, ANG II stimulated p42/p44 MAPK activation was not affected in hepatocytes obtained from rats fed ethanol chronically (6 weeks). However, chronic (6 weeks) ethanol treatment decreased ethanol potentiation of p42/p44 MAPK by about 56.3% +/- 3.6% for p42 MAPK and 61.3% +/- 11.7% for p44 MAPK. Furthermore, ethanol had no effect on the expression of angiotensinogen and c-myc mRNA in hepatocytes. A decrease in ANG II-activated phosphorylase a, but not in p42/p44 MAPK activation, after chronic (6 weeks) ethanol treatment leads to the conclusion that they may not be dependent on each other. PMID- 12782248 TI - Effects of ethanol or rimcazole on dizocilpine maleate-induced behaviors in male and female rats. AB - The current investigation was undertaken to explore further the interactions between ethanol and the phencyclidine analog dizocilpine maleate (MK-801) on behaviors in male and female rats. It was previously found that ethanol dependence conferred cross-tolerance to the behaviorally activating effects of dizocilpine. The current set of studies was designed to assay the interactions between dizocilpine and ethanol in ethanol-naive animals by measuring open field behaviors. I also tested interactions between dizocilpine and rimcazole, a sigma receptor antagonist. In agreement with previous reports, I found significant effects of dizocilpine on several open field behaviors. In general, female rats displayed a lower level of hyperlocomotion and higher level of stereotypies than did male rats. Co-administration of ethanol delayed time to peak hyperlocomotion in male rats. It reduced locomotion in female rats compared with findings for administration of dizocilpine alone. Co-administration of ethanol with dizocilpine increased stereotypies in both sexes. Administration of ethanol increased locomotion to a greater degree in female than in male rats. In contrast, co-administration of rimcazole with dizocilpine had little effect on hyperlocomotion in male rats while increasing levels in female rats. Rimcazole increased dizocilpine-induced stereotypies to a greater extent in male than in female rats. Results of receptor-binding studies revealed small differences for cerebral cortical sigma receptors between male and female rats. Dizocilpine was unable to compete for sigma receptor-binding sites. This is in contrast to phencyclidine, which acts at both N-methyl-D-aspartate (NMDA) and sigma receptors. These findings extend previous evidence of interactions between ethanol and dizocilpine, but highlight that responses vary by measure, sex, and length of ethanol exposure. In addition, findings from the current study uncovered sex-selective interactions between dizocilpine and a sigma receptor ligand, providing further evidence for complex actions and interactions of this noncompetitive NMDA receptor antagonist with multiple sites in brain. PMID- 12782250 TI - Interaction of aging and intermittent ethanol exposure on brain cytochrome c oxidase activity levels. AB - The effects of chronic, intermittent ethanol exposure on brain cytochrome c oxidase (CO) activity levels were studied in young (3- to 4-month-old) and aged (29- to 30-month-old) male Wistar rats. The rats were given highly intoxicating doses of ethanol three times a day by intragastric intubation for four successive days, followed by a 3-day ethanol-withdrawal period. This 4-day ethanol-exposure with 3-day ethanol-withdrawal cycle was repeated five times to simulate the binge drinking of human alcoholics. The histochemical demonstration of CO showed a markedly decreased activity level in the medial prefrontal cortex (especially layer V pyramids and neuropil) of the ethanol-exposed rats of both age groups compared with findings for the respective controls. In the cerebellar vermis, CO activity level was decreased in the Purkinje neurons of the aged ethanol-exposed rats and in the granule cells of both young and aged ethanol-exposed rats. The CO activity level in the locus coeruleus was decreased in both young and old ethanol exposed rats, but the decrease was more pronounced in the young ethanol-exposed group. Aging per se did not markedly change CO histochemical findings in either prefrontal or cerebellar cortex, but CO activity levels were increased in the locus coeruleus. In summary, results of the current study support our conclusion that CO activity levels were decreased in the cerebral and cerebellar cortices as well as in the locus coeruleus-CNS regions known to be negatively affected by chronic ethanol exposure. Defective energy metabolism due to decreased CO activity levels might compromise neuronal energy stores and thereby contribute to ethanol-induced brain dysfunction and irreversible CNS degeneration. PMID- 12782251 TI - Stable preference for high ethanol concentrations after ethanol deprivation in Sardinian alcohol-preferring (sP) rats. AB - Results of a recent study have demonstrated that exposure to multiple ethanol concentrations and repeated ethanol deprivation periods in Indiana ethanol preferring (P) rats resulted in the development of an alcohol deprivation effect (ADE; the temporary increase in voluntary ethanol intake after a period of deprivation from ethanol) characterized by consumption of intoxicating amounts of ethanol. The current study was designed to possibly extend these results to Sardinian alcohol-preferring (sP) rats, generated with the same selective program previously used for P rats. To this aim, ethanol-naive sP rats were exposed initially to the home cage four-bottle choice [10%, 20%, and 30% (vol./vol.) ethanol solutions and water] for eight consecutive weeks. Subsequently, rats were divided into two groups: The first group had continuous access to the four-bottle regimen (nondeprived rats), and the second group was exposed to five cycles of 14 day periods of deprivation from ethanol and 14-day periods of reexposure to the four-bottle regimen. An ADE developed after each deprivation period. However, the extra intake of ethanol was limited to the first hour of each reaccess period. Magnitude of ADE did not change with repeated periods of deprivation. However, a shift in preference toward the two highest concentrations of ethanol solutions was evident from the first reexposure to ethanol and was maintained throughout the study. These results provide further evidence on the heterogeneity of ethanol drinking behavior among rat lines selectively bred for high ethanol preference and consumption. PMID- 12782252 TI - Micro1-opioid antagonist naloxonazine alters ethanol discrimination and consumption. AB - The endogenous opioid system is implicated in excessive ethanol-drinking behavior. However, the role of individual opioid receptor subtypes in the mechanism underlying excessive ethanol-drinking behavior is not yet well understood. Therefore, we investigated the ability of a selective micro1-opioid antagonist, naloxonazine, to modulate ethanol-drinking behavior and ethanol discrimination in a rat model with the use of ethanol self-administration and drug discrimination paradigms. The effects of naloxonazine (0.001-10 mg/kg) on ethanol intake were examined in Sprague-Dawley rats under conditions of limited access to 10% (wt./vol.) ethanol and ad libitum access to food and water. Pretreatment with high doses of naloxonazine (1-10 mg/kg) significantly reduced ethanol consumption. When the effects of naloxonazine on food intake in free feeding male rats were examined, naloxonazine (1.8-10 mg/kg) significantly suppressed 24-h food intake. Another group of rats was trained to discriminate ethanol (1.25 g/kg, i.p.) from saline on a fixed-ratio schedule (FR 10), and ethanol dose-response tests were conducted once rats had acquired ethanol-saline discrimination. Injections were given 15 min before ethanol dose-response tests were conducted, and after characterization of the ethanol dose-response curve, the effects of naloxonazine on ethanol discrimination were assessed by administering naloxonazine (0.001-10 mg/kg, i.p.) 15 min before ethanol administration. Treatment with naloxonazine (0.001-1.8 mg/kg, i.p.) before the ED(100) dose of ethanol partially antagonized the discriminative stimulus of ethanol without having any effect on the response rate. The results support the suggestion of involvement of micro1-opioid receptors in the discriminative effects of ethanol and ethanol-drinking behavior. PMID- 12782253 TI - Extracellular levels of dopamine in the nucleus accumbens in AA and ANA rats after reverse microdialysis of ethanol into the nucleus accumbens or ventral tegmental area. AB - Ethanol is known to increase the release of dopamine in the nucleus accumbens. The question of whether this is a result of a direct or an indirect effect of ethanol on mesolimbic dopaminergic neurons was examined by investigating the extracellular levels of dopamine and its metabolites in the nucleus accumbens of alcohol-preferring AA (Alko Alcohol) and alcohol-avoiding ANA (Alko Non-Alcohol) rats after application of ethanol locally into either the nucleus accumbens or the ventral tegmental area with the use of reverse microdialysis. Application of ethanol (200, 400, or 800 mM in dialysate) into the nucleus accumbens, but not into the ventral tegmental area, temporarily increased the accumbal levels of dopamine in a dose-dependent manner. The ethanol-evoked increase in the level of extracellular dopamine was more prominent in AA rats than in ANA rats. Ethanol tended to suppress levels of 3,4-dihydroxyphenylacetic acid and homovanillic acid. Because the concentrations of ethanol found to elevate the extracellular level of dopamine can hardly be considered pharmacologically relevant, the increase in accumbal dopamine levels after application of ethanol may be due to nonspecific membrane effects of ethanol. The findings support the suggestion that the increase in the extracellular level of dopamine in the nucleus accumbens after systemic administration of ethanol may involve other sites on dopamine neurons or even different neurotransmitter systems, rather than the action of ethanol at the mesolimbic dopaminergic terminals. PMID- 12782254 TI - Nonlinear propagation model for ultrasound hydrophones calibration in the frequency range up to 100 MHz. AB - To facilitate the implementation and verification of the new ultrasound hydrophone calibration techniques described in the companion paper (somewhere in this issue) a nonlinear propagation model was developed. A brief outline of the theoretical considerations is presented and the model's advantages and disadvantages are discussed. The results of simulations yielding spatial and temporal acoustic pressure amplitude are also presented and compared with those obtained using KZK and Field II models. Excellent agreement between all models is evidenced. The applicability of the model in discrete wideband calibration of hydrophones is documented in the companion paper somewhere in this volume. PMID- 12782255 TI - Calibration of ultrasonic hydrophone probes up to 100 MHz using time gating frequency analysis and finite amplitude waves. AB - A number of ultrasound imaging systems employs harmonic imaging to optimize the trade off between resolution and penetration depth and center frequencies as high as 15 MHz are now used in clinical practice. However, currently available measurement tools are not fully adequate to characterize the acoustic output of such nonlinear systems primarily due to the limited knowledge of the frequency responses beyond 20 MHz of the available piezoelectric hydrophone probes. In addition, ultrasound hydrophone probes need to be calibrated to eight times the center frequency of the imaging transducer. Time delay spectrometry (TDS) is capable of providing transduction factor of the probes beyond 20 MHz, however its use is in practice limited to 40 MHz. This paper describes a novel approach termed time gating frequency analysis (TGFA) that provides the transduction factor of the hydrophone probes in the frequency domain and significantly extends the quasi-continuous calibration of the probes up to 60 MHz. The verification of the TGFA data was performed using TDS calibration technique (up to 40 MHz) and a nonlinear calibration method (up to 100 MHz). The nonlinear technique was based on a novel wave propagation model capable of predicting the true pressure-time waveforms at virtually any point in the field. The spatial averaging effects introduced by the finite aperture hydrophones were also accounted for. TGFA calibration results were obtained for different PVDF probes, including needle and membrane designs with nominal diameters from 50 to 500 micro m. The results were compared with discrete calibration data obtained from an independent national laboratory and the overall uncertainty was determined to be +/-1.5 dB in the frequency range 40-60 MHz and less than +/-1 dB below 40 MHz. PMID- 12782256 TI - A power ultrasonic technology for deliquoring. AB - Solid-liquid separation is a topic of permanent interest in many areas such as mineral recovery, food processing and sewage disposal. The adequate application of high-intensity ultrasonic fields may contribute to improve the efficiency of conventional deliquoring processes. Deliquoring refers to removal of liquid from a product without changing its phase. Different effects are involved in the application of high-intensity ultrasound for deliquoring, the most important of which are: the alternating acoustic stresses, the radiation pressure, the acoustic streaming, interface instabilities and cavitation. One of the main advantages of the ultrasonic energy in deliquoring processes is its ability to release the strongly bound moisture. This paper deals with the application of an ultrasonic procedure for deliquoring of slurries in which a high-intensity vibration, homogeneously distributed, is directly applied to the wet particulate material. The vibration travels through the solid-liquid medium and the rapid series of alternative compressions and rarefactions produce a kind of "sponge effect" which favours the migration of moisture through natural or acoustically created channels. The obtained results show that the new technique is very promising to assist filtration processes for solid-liquid separation of highly concentrated suspensions of fine particles. PMID- 12782257 TI - New technology for the design of advanced ultrasonic transducers for high-power applications. AB - A new high-frequency ultrasonic transducer for wire bonding has been conceived, designed, prototyped and tested. In the design phase an advanced approach was used and established. The method is based on the two basic principles of modularity and iteration. The transducer is decomposed to its elementary components. For each component an initial design is obtained with finite elements method (FEM) simulations. The simulated ultrasonic modules are then built and characterized experimentally through laser-interferometry measurements and electrical resonance spectra. The comparison of simulation results with experimental data allows the parameters of FEM models to be iteratively adjusted and optimized. The achieved FEM simulations exhibit a remarkably high-predictive potential and allow full control on the vibration behavior of the ultrasonic modules and of the whole transducer. The new transducer is fixed on the wire bonder with a flange whose special geometry was calculated by means of FEM simulations. This flange allows the converter to be attached on the wire bonder not only in longitudinal nodes but also in radial nodes of the ultrasonic field excited in the horn. This leads to a nearly complete decoupling of the transducer to the wire bonder, which has not been previously obtained. The new approach to mount ultrasonic transducers on a welding-device is of major importance not only for wire bonding but also for all high-power ultrasound applications and has been patented. PMID- 12782258 TI - Elastic friction drive of surface acoustic wave motor. AB - Importance of elastic deformation control to obtain large output force with a surface acoustic wave (SAW) motor is discussed in this paper. By adding pre-load to slider, stator and slider surfaces are deformed in a few tens nanometer. Appropriate deformation in normal direction against normal vibration displacement amplitude of SAW existed. By moderate deformation, the output force of the SAW motor was enlarged up to about 10 N and no-load speed was 0.7 m/s. To produce this performance, the transducer weight and slider size were only 4.2 g and 4 x 4 mm(2).By traveling wave propagation, surface particles of the SAW device move in elliptical motion. Due to the amplitude of the elliptical motion is 10 or 20 nm order, the contact condition of the slider is very critical. To control the contact condition, namely, the elastic deformation of the slider and stator surface in nanometer order, a lot of projections were fabricated on the slider surface. The projection diameter was 20 micro m. In static condition, the elastic deformation and stress were evaluated with the FEM analysis. From this calculation and the simulation result, it is consider that the wave crest is distorted, hence the elasticity has influence on the friction drive condition. Elastic deformation of the stator surface beneath the projection from the initial position were evaluated. In 4 x 4 mm(2) square area, the sliders had from 1089 to 23,409 projections. Depression was independent to the contact pressure. However, the output force depended on the depression although the projection density were different. From the view point of the output power of the motor, the proper depression was independent to the projection density. Around 25 nm depression, the output force and output power were maximized. This depression value was almost same as the vibration displacement amplitude of the stator transducer. PMID- 12782260 TI - The design of a high power ultrasonic test cell using finite element modelling techniques. AB - This paper will describe the application of a finite element (FE) code to design a test cell, in which a single transducer is used to generate acoustic cavitation. The FE model comprises a 2-D slice through the centre of the test cell and was used to evaluate the generated pressure fields as a function of frequency. Importantly, the pressure fields predicted by FE modelling are used to indicate the position of pressure peaks, or 'hot-spots', and nulls enabling the systems design engineer to visualise both the potential cavitation areas, corresponding to the 'hot-spots', and areas of low acoustic pressure. Through this design process, a rectangular test cell was constructed from perspex for use with a 40 kHz Tonpilz transducer. A series of experimental measurements was conducted to evaluate the cavitation threshold as a function of temperature and viscosity/surface tension, for different fluid load media. The results indicate the potential of the FE design approach and assist the design engineer in understanding the influence of the fluid load medium on the cell's ability to produce a strong cavitation field. PMID- 12782259 TI - Investigation of the influence of humidity on the ultrasonic agglomeration of submicron particles in diesel exhausts. AB - Removing very fine particles in the 0.01-1 micro m range generated in diesel combustion is important for air pollution abatement because of the impact such particles have on the environment. By forming larger particles, acoustic agglomeration of submicron particles is presented as a promising process for enhancing the efficiency of the current filtration systems for particle removal. Nevertheless, some authors have pointed out that acoustic agglomeration is much more efficient for larger particles than for smaller particles. This paper studies the effect of humidity on the acoustic agglomeration of diesel exhausts particles in the nanometer size range at 21 kHz. For the agglomeration tests, the experimental facility basically consists of a pilot scale plant with a diesel engine, an ultrasonic agglomeration chamber a dilution system, a nozzle atomizer, and an aerosol sampling and measuring station. The effect of the ultrasonic treatment, generated by a linear array of four high-power stepped-plate transducers on fumes at flow rates of 900 Nm(3)/h, was a small reduction in the number concentration of particles at the outlet of the chamber. However, the presence of humidity raised the agglomeration rate by decreasing the number particle concentration by up to 56%. A numerical study of the agglomeration process as a linear combination of the orthokinetic and hydrodynamic agglomeration coefficients resulting from mutual radiation pressure also found that acoustic agglomeration was enhanced by humidity. Both results confirm the benefit of using high-power ultrasound together with humidity to enhance the agglomeration of particles much smaller than 1 micro m. PMID- 12782261 TI - Enhancement of gas phase heat transfer by acoustic field application. AB - This study discusses a possibility for enhancement of heat transfer between solids and ambient gas by application of powerful acoustic fields. Experiments are carried out by using preheated Pt wires (length 0.1-0.15 m, diameter 50 and 100 micro m) positioned at the velocity antinode of a standing wave (frequency range 216-1031 Hz) or in the path of a travelling wave (frequency range 6.9-17.2 kHz). A number of experiments were conducted under conditions of gas flowing across the wire surface. Effects of sound frequency, sound strength, gas flow velocity and wire preheating temperature on the Nusselt number are examined with and without sound application. The gas phase heat transfer rate is enhanced with acoustic field strength. Higher temperatures result in a vigorous radiation from the wire surface and attenuate the effect of sound. The larger the gas flow velocity, the smaller is the effect of sound wave on heat transfer enhancement. PMID- 12782262 TI - All-optical adaptive scanning acoustic microscope. AB - We have constructed a fast laser-based surface acoustic wave (SAW) microscope, which may be thought of as a non-perturbing scanning acoustic microscope. The instrument is capable of rapid high resolution vector contrast imaging at several discrete frequencies, without any damage to the sample. Tailoring the generating optical distribution using computer-generated holograms allows us to both focus the acoustic waves (increasing their amplitude) and to spread the optical power over the sample surface (preventing damage). Accurate quantitative amplitude and phase (velocity) measurements and unique acoustic contrast mechanisms are possible with our instrument based on this technology due to the non-perturbing nature and the instrument geometries.However, the complexity of the optical generation profile leads to a strong dependence on material properties such as the SAW velocity and material anisotropy. We address these issues in this paper, and demonstrate how a spatial light modulator may be used to adapt the generating optical distribution to compensate for the material properties. This facilitates simpler alignment and velocity matching, and, combined with an acoustic wavefront sensor, will allow real-time adjustment of the generating source to enable imaging on anisotropic materials. PMID- 12782263 TI - Hidden corrosion detection in aircraft aluminum structures using laser ultrasonics and wavelet transform signal analysis. AB - Preliminary results of hidden corrosion detection in aircraft aluminum structures using a noncontact laser based ultrasonic technique are presented. A short laser pulse focused to a line spot is used as a broadband source of ultrasonic guided waves in an aluminum 2024 sample cut from an aircraft structure and prepared with artificially corroded circular areas on its back surface. The out of plane surface displacements produced by the propagating ultrasonic waves were detected with a heterodyne Mach-Zehnder interferometer. Time-frequency analysis of the signals using a continuous wavelet transform allowed the identification of the generated Lamb modes by comparison with the calculated dispersion curves. The presence of back surface corrosion was detected by noting the loss of the S(1) mode near its cutoff frequency. This method is applicable to fast scanning inspection techniques and it is particularly suited for early corrosion detection. PMID- 12782264 TI - Nonlinear characterization with burst excitation of 1-3 piezocomposite transducers. AB - Ultrasonic transducers made with 1-3 connectivity piezocomposites are frequently used in Medical applications and nondestructive testing. When the transducer is used for special applications as, for instance air-coupled transmission, it is necessary to compensate for the high difference of acoustic impedance between transducer and medium using high amplitude pulses to generate high acoustic signal. Thus, the nonlinear behavior of the transducer must be taken into account in similar application conditions. The newly developed method, which performs the nonlinear characterization with burst signal excitation near the thickness resonance frequency, is based on the measure of the current as well as the vibration velocity of the piezocomposite transducer. The current of the stationary response is measured before the end of the burst signal excitation. Burst excitation enables us to measure the nonlinear characterization without producing overheating in the transducers. The amplitude level dependence of mechanical losses tandelta(m) and the stiffness increases |Deltac/c(0)| have been studied, as well as the velocity dependence of a point of the transducer, measured with a laser vibrometer. In this method, the power level applied to the transducers can be higher than other nonlinear measurement methods, providing measurements of high accuracy. PMID- 12782265 TI - The cavitation induced Becquerel effect and the hot spot theory of sonoluminescence. AB - Over 150 years ago, Becquerel discovered the ultraviolet illumination of one of a pair of identical electrodes in liquid water produced an electric current, the phenomenon called the Becquerel effect. Recently, a similar effect was observed if the water surrounding one electrode is made to cavitate by focused acoustic radiation, which by similarity is referred to as the cavitation induced Becquerel effect. The current in the cavitation induced Becquerel effect was found to be semi-logarithmic with the standard electrode potential that is consistent with the oxidation of the electrode surface by the photo-decomposition theory of photoelectrochemistry. But oxidation of the electrode surface usually requires high temperatures, say as in cavitation. Absent high bubble temperatures, cavitation may produce vacuum ultraviolet (VUV) light that excites water molecules in the electrode film to higher H(2)O(*) energy states, the excited states oxidizing the electrode surface by chemical reaction. Solutions of the Rayleigh-Plesset equation during bubble collapse that include the condensation of water vapor show any increase in temperature or pressure of the water vapor by compression heating is compensated by the condensation of vapor to the bubble wall, the bubbles collapsing almost isothermally. Hence, the cavitation induced Becquerel effect is likely caused by cavitation induced VUV light at ambient temperature. PMID- 12782266 TI - Liquid level sensor using ultrasonic Lamb waves. AB - This paper describes a novel, noninvasive method for measurement of liquid level in closed metal tanks that are under high pressure. It is based on the use of ultrasonic Lamb waves propagating along the tank wall. Contact with liquid substantially changes the characteristics of these waves and this can be used as an indicator of liquid presence. Theoretical analysis shows that the symmetric and antisymmetric Lamb wave modes, both fundamental and higher order, are sensitive to presence of the liquid. The optimal wave frequency depends on the thickness of the tank wall and wall material. A prototype level sensor based on this principle has been developed. It uses two pairs of wedge transducers to generate and detect Lamb waves propagating along the circumference of the gas tank. An operating frequency of 100 kHz is found to be optimal for use with tanks having a wall thickness of 30-50 mm. Prototype sensors developed under this program have been used successfully in oil fields in the far northern region of Russia. PMID- 12782267 TI - Cavitation induced Becquerel effect. AB - The observation of an electrical current upon the ultraviolet (UV) illumination of one of a pair of identical electrodes in liquid water, called the Becquerel effect, was made over 150 years ago. More recently, an electrical current was found if the water surrounding one electrode was made to cavitate by focused acoustic radiation, the phenomenon called the cavitation induced Becquerel effect. Since cavitation is known to produce UV light, the electrode may simply absorb the UV light and produce the current by the photo-emission theory of photoelectrochemistry. But the current was found to be semi-logarithmic with the standard electrode potential which is characteristic of the oxidation of the electrode surface in the photo-decomposition theory, and not the photo-emission theory. High bubble collapse temperatures may oxidize the electrode, but this is unlikely because melting was not observed on the electrode surfaces. At ambient temperature, oxidation may proceed by chemical reaction provided a source of vacuum ultraviolet (VUV) radiation is available to produce the excited OH* states of water to react with the electrode. The source of VUV radiation is shown to be the spontaneous emission of coherent infrared (IR) radiation from water molecules in particles that form in bubbles because of surface tension, the spontaneous IR emission induced by cavity quantum electrodynamics. The excited OH* states are produced as the IR radiation accumulates to VUV levels in the bubble wall molecules. PMID- 12782268 TI - Ultrasonic-trap-enhanced selectivity in capillary electrophoresis. AB - We combine ultrasonic trapping and capillary electrophoresis (CE) with the goal to detect ultra-low concentrations of proteins via size-selective separation and enrichment of antibody-coated latex spheres. An 8.5 MHz standing ultrasonic wave is longitudinally coupled into the sub-100- micro m diam capillary of the CE system. Competition between acoustic and viscous forces result in in-flow separation of micro m-diam spheres according to their size. Experiments separating 2.8- and 2.1- micro m-diam fluorescent latex particles, which model a protein-specific immunocomplex/free particle mixture, indicate a potential improvement of the concentration limit of detection of 10(4) compared to current CE systems. Theoretical calculations show room for further improvement. PMID- 12782269 TI - Prepattern genes and signaling molecules regulate stripe expression to specify Drosophila flight muscle attachment sites. AB - In Drosophila, muscles attach to epidermal tendon cells specified by the gene stripe (sr). Flight muscle attachment sites are prefigured on the wing imaginal disc by sr expression in discrete domains. We describe the mechanisms underlying the specification of these domains of sr expression. We show that the concerted activities of the wingless (wg), decapentaplegic (dpp) and Notch (N) signaling pathways, and the prepattern genes pannier (pnr) and u-shaped (ush) establish domains of sr expression. N is required for initiation of sr expression. pnr is a positive regulator of sr, and is inhibited by ush in this function. The Wg signal differentially influences the formation of different sr domains. These results identify the multiple regulatory elements involved in the positioning of Drosophila flight muscle attachment sites. PMID- 12782270 TI - Vein-positioning in the Drosophila wing in response to Hh; new roles of Notch signaling. AB - The Drosophila wing is a classical model for studying the generation of developmental patterns. Previous studies have suggested that vein primordia form at boundaries between discrete sectors of gene expression along the antero posterior (A/P) axis in the larval wing imaginal disc. Observation that the vein marker rhomboid (rho) is expressed at the centre of wider vein-competent domains led to propose that narrow vein primordia form first, and produce secondary short range signals activating provein genes in neighbouring cells (see Curr. Opin. Genet. Dev. 10 (2000) 393). Here, we examined how the central L3 and L4 veins are positioned relative to the limits of expression of Collier (Col), a dose dependent Hedgehog (Hh) target activated in the wing A/P organiser. We found that rho expression is first activated in broad domains adjacent to Col-expressing cells and secondarily restricted to the centre of these domains. This restriction which depends upon Notch (N) signaling sets the L3 and L4 vein primordia off the boundaries of Col expression. N activity is also required to fix the anterior limit of Col expression by locally antagonising Hh activation, thus precisely positioning the L3 vein primordium relative to the A/P compartment boundary. Experiments using Nts mutants further indicated that these two activities of N could be temporally uncoupled. Together, these observations highlight new roles of N in topologically linking the position of veins to prepattern gene expression. PMID- 12782271 TI - Requirement of the roughest gene for differentiation and time of death of interommatidial cells during pupal stages of Drosophila compound eye development. AB - The roughest locus of Drosophila melanogaster encodes a transmembrane protein of the immunoglobulin superfamily required for several developmental processes, including axonal pathfinding in the developing optic lobe, mechanosensory bristle differentiation and myogenesis. In the compound eye, rst was previously shown to be required for establishing the correct number and spacing of secondary and tertiary pigment cells during the final steps of ommatidial assembly. We have further investigated its function in the developing pupal retina by performing a developmental and molecular analysis of a novel dominant rst allele, rst(D). In addition to showing evidence that rst(D) is a regulatory mutant, the results strongly suggest a previously unnoticed role of the rst gene in the differentiation of secondary/tertiary pigment cell fate as well as establishing the correct timing of surplus cell removal by programmed cell death in the compound eye. PMID- 12782272 TI - Genetic ablation of the tumor suppressor menin causes lethality at mid-gestation with defects in multiple organs. AB - Patients suffering from multiple endocrine neoplasia type 1 (MEN1) are predisposed to multiple endocrine tumors. The MEN1 gene product, menin, is expressed in many embryonic, as well as adult tissues, and interacts with several proteins in vitro and in vivo. However, the biological function of menin remains largely unknown. Here we show that disruption of the Men1 gene in mice causes embryonic lethality at E11.5-E13.5. The Men1 null mutant embryos appeared smaller in size, frequently with body haemorrhages and oedemas, and a substantial proportion of them showed disclosure of the neural tube. Histological analysis revealed an abnormal development of the nervous system and heart hypotrophy in some Men1 null embryos. Furthermore, Men1 null livers generally displayed an altered organization of the epithelial and hematopoietic compartments associated with enhanced apoptosis. Chimerism analysis of embryos generated by injection of Men1 null ES cells, showed that cells lacking menin do not seem to have a general cell-autonomous defect. However, primary Men1 null embryonic fibroblasts entered senescence earlier than their wild-type counterparts. Despite normal proliferation ability, Men1 null ES cells exhibited a deficiency to form embryoid bodies, suggesting an impaired differentiation capacity in these cells. The present study demonstrates that menin plays an important role in the embryonic development of multiple organs in addition to its proposed role in tumor suppression. PMID- 12782273 TI - Coquillette, a sea urchin T-box gene of the Tbx2 subfamily, is expressed asymmetrically along the oral-aboral axis of the embryo and is involved in skeletogenesis. AB - Transcription factors of the T-domain family regulate many developmental processes. We have isolated from the sea urchin a new member of the Tbx2 subfamily: coquillette. Coquillette has a late zygotic expression whose localization is dynamic: at the blastula stage it is restricted to the aboral side of most of the presumptive ectoderm and endoderm territories and from gastrulation on, to the aboral-most primary mesenchyme cells. Perturbation of coquillette function delays gastrulation and strongly disorganizes the skeleton of the larva. Coquillette is sensitive to alteration of the oral-aboral (OA) axis and we identify goosecoid, which controls oral and aboral fates in the ectoderm, as a probable upstream regulator. Coquillette appears to be an integral part of the patterning system along the OA axis. PMID- 12782274 TI - Vertebrate caudal gene expression gradients investigated by use of chick cdx A/lacZ and mouse cdx-1/lacZ reporters in transgenic mouse embryos: evidence for an intron enhancer. AB - The vertebrate caudal proteins, being upstream regulators of the Hox genes, play a role in establishment of the body plan. We describe analysis of two orthologous caudal genes (chick cdx-A and mouse cdx-1) by use of lacZ reporters expressed in transgenic mouse embryos. The expression patterns show many similarities to the expression of endogenous mouse cdx-1. At 8.7 days, cdx/lacZ activity within neurectoderm and mesoderm forms posterior-to-anterior gradients, and we discuss the possibility that similar gradients of cdx gene expression may function as morphogen gradients for the establishment of Hox gene expression boundaries. Our observations suggest that gradients form by decay of cdx/lacZ activity in cells that have moved anterior to the vicinity of the node. The cdx-A/lacZ expression pattern requires an intron enhancer that includes two functional control elements: a DR2-type retinoic acid response element and a Tcf/beta-catenin binding motif. These motifs are structurally conserved in mouse cdx-1. PMID- 12782275 TI - Further developmental roles of the Vestigial/Scalloped transcription complex during wing development in Drosophila melanogaster. AB - The Drosophila homologue of the human TEF-1 gene, scalloped (sd), is required for wing development. The SD protein forms part of a transcriptional activation complex with the protein encoded by vestigial (vg) that, in turn, activates target genes important for wing formation. One sd function involves a regulatory feedback loop with vg and wingless (wg) that is essential in this process. The dorsal-ventral (D/V) margin-specific expression of wg is lost in sd mutant wing discs while the hinge-specific expression appears normal. In the context of wing development, a VG::sdTEA domain fusion produces a protein that mimics the wild type SD/VG complex and restores the D/V boundary-specific expression of wg in a sd mutant background. Further, targeted expression of wg at the D/V boundary in the wing disc was able to partially rescue the sd mutant phenotype. This infers that sd could function in either the maintenance or induction of wg at the D/V border. Another functional role for sd is the establishment of sensory organ precursors (SOP) of the peripheral nervous system at the wing margin. Thus, the relationship between sd and senseless (sens) in the development of these cells is also examined, and it appears that sd must be functional for proper sens expression, and ultimately, for sensory organ precursor development. PMID- 12782276 TI - Asymmetric limb malformations in a new transgene insertional mutant, footless. AB - Six to eight copies of a transgene integrated into mouse chromosome 15 resulting in a new transgene insertional mutant, Footless, presenting with malformations of the limbs, kidney, and soft palate. Homozygotes possess a unique asymmetric pattern of limb truncations. Posterior structures from the autopod and zeugopod of the hindlimbs are missing with left usually more severely affected than right. In contrast, anterior structures are missing from the right forelimbs. The left forelimb is usually normal except for the absence of the distal telephalanges and nails. These structures are absent on all formed digits. In situ hybridization assays examined the expression of Shh, dHand, Msx2, Fgf8, En1, and Lmx1b in mutant limb buds and indicated normal establishment of the anterior/posterior and dorsal/ventral axes of the developing limbs. However, dysmorphology of the apical ectodermal ridge was observed in the mutant limb buds. PMID- 12782277 TI - Depletion of the cell-cycle inhibitor p27(Xic1) impairs neuronal differentiation and increases the number of ElrC(+) progenitor cells in Xenopus tropicalis. AB - The Xenopus p27(Xic1) gene encodes a cyclin dependent kinase (CDK) inhibitor of the Cip/Kip family. We have previously shown that p27(Xic1) is expressed in the cells of the neural plate as they become post-mitotic (Development 127 (2000) 1303). To investigate whether p27(Xic1) is necessary for cell cycle exit and/or neuronal differentiation, we used antisense morpholino oligos (MO) to knockdown the protein levels in vivo. For such knockdown studies, Xenopus tropicalis is a better model system than Xenopus laevis, since it has a diploid genome. Indeed, while X. laevis has two p27(Xic1) paralogs, p27(Xic1) and p28(Kix1), we have found only one ortholog in X. tropicalis, equidistant from the X. laevis genes. The X. tropicalis p27(Xic1) was expressed in a similar pattern to the X. laevis gene. Depletion of p27(Xic1) in X. tropicalis caused an increase in proliferation and a suppression of the neuronal differentiation marker, N-tubulin. At the same time, we found an increase in the expression of ElrC, a marker of cells as they undergo a transition from proliferation to differentiation. We conclude that p27(Xic1) is necessary for cells to exit the cell cycle and differentiate; in its absence, cells accumulate in a progenitor state. The expression of p27(Xic1) in the embryo is regionalised but the transcriptional regulation of p27(Xic1) is not well understood. We report the isolation of a p27(Xic1) genomic clone and we identify a 5' region capable of driving reporter gene expression specifically in the neural tube and the eye. PMID- 12782278 TI - L63, the Drosophila PFTAIRE, interacts with two novel proteins unrelated to cyclins. AB - L63 encodes a CDK-like protein homologous to the mammalian PFTAIRE. We showed previously that L63 provides a CDK-related function critical to development (Dev. Biol. 221 (2000) 23). We present here the first biochemical characterization of L63 kinase. In addition, we describe two novel Drosophila proteins, PIF-1 and PIF 2 (for PFTAIRE Interacting Factor-1 and -2), identified in a two-hybrid screen for their ability to interact with the amino-terminal region of L63. The full length PIF-1 cDNA shows an unusual dicistronic organization. PIF-1A and PIF-1B (the L63 interactor) predicted proteins are expressed in vivo, and show a distinct expression profile during development. Interaction between L63 and PIF 1B was confirmed in vitro and in vivo. The role of this interaction remains to be demonstrated, but our data suggest that PIF-1B might serve as a regulator of L63. PMID- 12782279 TI - Tob proteins enhance inhibitory Smad-receptor interactions to repress BMP signaling. AB - Tob inhibits bone morphogenetic protein (BMP) signaling by interacting with receptor-regulated Smads in osteoblasts. Here we provide evidence that Tob also interacts with the inhibitory Smads 6 and 7. A yeast two-hybrid screen identified Smad6 as a protein interacting with Tob. Tob co-localizes with Smad6 at the plasma membrane and enhances the interaction between Smad6 and activated BMP type I receptors. Furthermore, we have isolated Xenopus Tob2, and show that it cooperates with Smad6 in inducing secondary axes when expressed in early Xenopus embryos. Finally, Tob and Tob2 cooperate with Smad6 to inhibit endogenous BMP signaling in Xenopus embryonic explants and in cultured mammalian cells. Our results provide both in vitro and in vivo evidence that Tob inhibits endogenous BMP signaling by facilitating inhibitory Smad functions. PMID- 12782280 TI - DNA: the double helix. PMID- 12782281 TI - Microbial isoprenoid biosynthesis and human gammadelta T cell activation. AB - Human Vgamma9/Vdelta2 T cells play a crucial role in the immune response to microbial pathogens, yet their unconventional reactivity towards non-peptide antigens has been enigmatic until recently. The break-through in identification of the specific activator was only possible due to recent success in a seemingly remote field: the elucidation of the reaction steps of the newly discovered 2-C methyl-D-erythritol-4-phosphate (MEP) pathway of isoprenoid biosynthesis that is utilised by many pathogenic bacteria. Unexpectedly, the intermediate of the MEP pathway, (E)-4-hydroxy-3-methyl-but-2-enyl-pyrophosphate) (HMB-PP), turned out to be by far the most potent Vgamma9/Vdelta2 T cell activator known, with an EC(50) of 0.1 nM. PMID- 12782282 TI - Conserved hydrophobicity in the SH2-kinase linker is required for catalytic activity of Csk and CHK. AB - The crystal structure of full-length Csk (C-terminal Src kinase) molecules shows a hydrophobic interaction between the SH2-kinase linker residue Phe183 and the alphaC-helix of the catalytic domain. To study the possible involvement of this contact in the regulation of the activity of Csk and CHK (Csk homologous kinase), a Csk SH2-kinase linker deletion mutant, Csk Phe183 and CHK Leu223 point mutants were analyzed. It was observed that a residue with a long hydrophobic side chain in position 183 (Csk) and 223 (CHK) is required to sustain the catalytic activity of Csk and CHK. These results suggest that Csk Phe183 and CHK Leu223 stabilize the movement of the alphaC-helix of these protein tyrosine kinases. PMID- 12782283 TI - EPR study of light-induced regulation of photosynthetic electron transport in Synechocystis sp. strain PCC 6803. AB - The kinetics of the light-induced redox changes of the photosystem 1 (PS 1) primary donor P(700) in whole cells of the cyanobacteria Synechocystis sp. PCC 6803 were studied by the electron paramagnetic resonance method. It was shown that the linear photosynthetic electron transport in cyanobacteria was controlled by two main mechanisms: (i) oxygen-dependent acceleration of electron transfer from PS 1 to NADP(+) due to activation of the Calvin cycle reactions and (ii) retardation of electron flow between two photosystems governed by a transmembrane proton gradient. In addition to the linear photosynthetic electron transport, cyanobacteria were capable of maintaining alternative pathways involving cyclic electron transfer around PS 1 and respiratory chains. PMID- 12782284 TI - Effects of pathological mutations on the stability of a conserved amino acid triad in retinoschisin. AB - A three-dimensional model has been calculated for the discoidin domain of retinoschisin (RS1), the protein involved in the X-linked juvenile retinoschisis. The model allows for a mapping of the pathological retinoschisis missense mutations and a rationale for the structural effects of an evolutionary conserved surface exposed triad (W122-R200-W163). Molecular dynamics simulations of the triad mutants models, together with ab initio energy calculations of the complexes corresponding to the triad show that the observed pathological mutations sensibly destabilize local interactions and the entire fold. Moreover the presented model reveals evidence of a putative site for membrane association. PMID- 12782285 TI - The human G protein beta4 subunit: gene structure, expression, Ggamma and effector interaction. AB - The aim of this study was the characterization of the human Gbeta4 subunit of heterotrimeric G proteins. Human Gbeta4 is widely expressed. Its gene is located on chromosome 3 with a genomic structure indistinguishable from that of the genes of Gbeta1 to Gbeta3, but entirely different from Gbeta5. In vitro translation co precipitation analyses revealed that Gbeta4 can form stable dimers with Ggamma1, Ggamma2, Ggamma3, Ggamma4, Ggamma5, Ggamma7, Ggamma10, Ggamma11, Ggamma12, and Ggamma13, dimers which were also able to stimulate phospholipase beta2. PMID- 12782286 TI - Critical regions for the sweetness of brazzein. AB - Brazzein is a small, heat-stable, intensely sweet protein consisting of 54 amino acid residues. Based on the wild-type brazzein, 25 brazzein mutants have been produced to identify critical regions important for sweetness. To assess their sweetness, psychophysical experiments were carried out with 14 human subjects. First, the results suggest that residues 29-33 and 39-43, plus residue 36 between these stretches, as well as the C-terminus are involved in the sweetness of brazzein. Second, charge plays an important role in the interaction between brazzein and the sweet taste receptor. PMID- 12782287 TI - Transcriptional repression of cyclin-dependent kinase inhibitor p21 gene by phospholipase D1 and D2. AB - Phospholipase D (PLD) is known to stimulate cell cycle progression and to transform murine fibroblast cells into tumorigenic forms, although the precise mechanisms are not elucidated. In this report, we demonstrated that both PLD1 and PLD2 repressed expression of cyclin-dependent kinase inhibitor p21 gene in an additive manner. The phospholipase activity of PLDs was important for the effect. PLD1 repressed the p21 promoter by decreasing the level of p53, whereas PLD2 via a p53-independent pathway through modulating Sp1 activity. Taken together, we suggest that PLD isozymes stimulate cell growth by repressing expression of p21 gene, which may ultimately lead to carcinogenesis. PMID- 12782288 TI - Met174 side chain is the site of photoinsertion of a substance P competitive peptide antagonist photoreactive in position 8. AB - Numerous photoaffinity studies of the NK-1 receptor have been carried out with peptide agonist analogues of substance P (SP). However, no information is available with regard to the domain interaction of peptide antagonists within this receptor. We describe herein the photoaffinity labelling of the SP receptor with a peptide antagonist analogue, Bapa(0)[(pBzl)Phe(8),DPro(9),MePhe(10),Trp(CHO)(11)]SP. Photolabelling, enzymatic or chemical cleavage of the covalent complex, purification via streptavidin coated beads and matrix-assisted laser desorption/ionization time of flight mass spectrometry analysis led us to show that the methyl of Met174 side chain, within the receptor's second extracellular loop, is covalently linked to the antagonist photoreactive at position 8. PMID- 12782289 TI - NMR studies of the fifth transmembrane segment of sarcoplasmic reticulum Ca2+ ATPase reveals a hinge close to the Ca2+-ligating residues. AB - Two recent X-ray structures have tremendously increased the understanding of the sarco/endoplasmic reticulum Ca(2+)-ATPase (SERCA) and related proteins. Both structures show the fifth transmembrane span (M5) as a single continuous alpha helix. The inherent structural and dynamic features of this span (Lys758-Glu785) were studied in isolation in sodium dodecyl sulfate (SDS) micelles using liquid state nuclear magnetic resonance (NMR) spectroscopy. We find that a flexible region (Ile765-Asn768) is interrupting the alpha-helix. The location of the flexible region near the Ca(2+) binding residues Asn768 and Glu771 suggests that together with a similar region in M6 it has a hinge function that may be important for cooperative Ca(2+) binding and occlusion. PMID- 12782290 TI - Palmitoylation sites and processing of synaptotagmin I, the putative calcium sensor for neurosecretion. AB - Synaptotagmin I, the calcium sensor for neurotransmission, is palmitoylated. We have identified the palmitoylation sites as five cysteine residues located between the transmembrane and cytoplasmic regions. In contrast to wild-type synaptotagmin, the non-acylated mutant is not converted to the endoglycosidase-H resistant form after expression in CV-1 cells. This indicates a block in transport through the Golgi complex. However, when expressed in PC-12 and RBL cells non-acylated synaptotagmin is targeted to the plasma membrane and to secretory granules. No significant cleavage of [(3)H]palmitate from synaptotagmin was observed in pulse-chase experiments. This indicates that the majority of fatty acids are structural rather than dynamic components. PMID- 12782291 TI - Transient increase of phosphatidylcholine in plant cells in response to phosphate deprivation. AB - In plants, phosphate deprivation is normally known to decrease the phospholipid content consistent with a mobilization of the phosphate reserve, and conversely to increase non-phosphorous membrane lipids such as digalactosyldiacylglycerol. We report here that unexpectedly, at an early stage of phosphate starvation, phosphatidylcholine (PC) increases transiently. We also show that a significant pool of diacylglycerol (DAG) with the same fatty acid composition as that of PC is present and moreover increases in response to phosphate deprivation. The evolution of the molecular profile of the newly synthesized galactolipids is compatible with a utilization of DAG accumulating from PC hydrolysis, achieved after selection of their acyl molecular species by the galactolipid synthesizing enzymes. PMID- 12782292 TI - Snorkeling of lysine side chains in transmembrane helices: how easy can it get? AB - Transmembrane segments of proteins are often flanked by lysine residues. The side chains of these residues may snorkel, i.e. they may bury themselves with their aliphatic part in the hydrophobic region of the lipid bilayer, while positioning the charged amino group in the more polar interface. Here we estimate the free energy cost of snorkeling from thermodynamical calculations based on studies with synthetic transmembrane peptides [Strandberg et al. (2002) Biochemistry 41, 7190 7198]. The value is estimated to be between 0.07 and 0.7 kcal mol(-1) for a lysine side chain. This very low value indicates that snorkeling may be a common process, which should be taken into consideration both in experimental and in theoretical studies on protein-lipid interactions. PMID- 12782293 TI - Growth phase-dependent changes in the subcellular localization of pre-B-cell colony-enhancing factor. AB - A cDNA encoding the homolog of the human pre-B-cell colony-enhancing factor (PBEF), a cytokine-like secreted protein, was isolated from a rat cDNA library. This protein existed in both the cytoplasm and nucleus of the cells, and the amount was higher in the cytoplasm than in the nucleus of proliferating PC-12 and Swiss 3T3 cells but higher in the nucleus than in the cytoplasm of the PC-12 cells treated with nerve growth factor and the 3T3 cells grown to a confluent state. Thus, the so-called PBEF is not a cytokine-like secreted protein but an intracellular protein associated with the cell cycle. PMID- 12782294 TI - Unusual nucleotide-binding properties of the chloroplast protein import receptor, atToc33. AB - Arabidopsis Toc33 (atToc33) is a GTP-binding protein of the chloroplast outer envelope membrane. We studied its nucleotide-binding properties in vitro, and found that it binds GTP, GDP and XTP, with similar efficiencies, but not ATP. We further demonstrated that atToc33 has intrinsic GTPase activity. Mutations within the putative G4 motif of the atToc33 nucleotide-binding domain (D217N, D219N and E220Q) had no effect on nucleotide specificity or GTPase activity. Similarly, a mutation in the newly assigned G5 motif (E208Q) did not affect nucleotide specificity or GTPase activity. Furthermore, the D217N and D219N mutations did not affect atToc33 functionality in vivo. The data demonstrate that atToc33 belongs to a novel class of GTPases with unusual nucleotide-binding properties. PMID- 12782295 TI - Adrenomedullin provokes endothelial Akt activation and promotes vascular regeneration both in vitro and in vivo. AB - We previously reported that adrenomedullin (AM), a vasodilating hormone secreted from blood vessels, promotes proliferation and migration of human umbilical vein endothelial cells (HUVECs). In this study, we examined the ability of AM to promote vascular regeneration. AM increased the phosphorylation of Akt in HUVECs and the effect was inhibited by the AM antagonists and the inhibitors for protein kinase A (PKA) or phosphatidylinositol 3-kinase (PI3K). AM promoted re endothelialization in vitro of wounded monolayer of HUVECs and neo vascularization in vivo in murine gel plugs. These effects were also inhibited by the AM antagonists and the inhibitors for PKA or PI3K. The findings suggest that AM plays significant roles in vascular regeneration, associated with PKA- and PI3K-dependent activation of Akt in endothelial cells, and possesses therapeutic potential for vascular injury and tissue ischemia. PMID- 12782296 TI - Divergent evolution of flavonoid 2-oxoglutarate-dependent dioxygenases in parsley. AB - Flavone synthases (FNSs) catalyze the oxidation of flavanones to flavones, i.e. the formation of apigenin from (2S)-naringenin. While many plants express a microsomal-type FNS II, the soluble FNS I appears to be confined to a few species of the Apiaceae and was cloned recently from parsley plants. FNS I belongs to the Fe(II)/2-oxoglutarate-dependent dioxygenases characterized by short conserved sequence elements for cofactor binding, and its evolutionary context and mode of action are under investigation. Using a homology-based reverse transcription polymerase chain reaction approach, two additional flavonoid-specific dioxygenases were cloned from immature parsley leaflets, which were identified as flavanone 3beta-hydroxylase (FHT) and flavonol synthase (FLS) after expression in yeast cells. Sequence alignments revealed marginal differences among the parsley FNS I and FHT polypeptides of only 6%, while much less identity (about 29%) was observed with the parsley FLS. Analogous to FNS I, FLS oxidizes the flavonoid gamma-pyrone by introducing a C2, C3 double bond, and (2R,3S)-dihydrokaempferol (cis-dihydrokaempferol) was proposed recently as the most likely intermediate in both FNS I and FLS catalysis. Incubation of either FNS I or FLS with cis dihydrokaempferol exclusively produced kaempferol and confirmed the assumption that flavonol formation occurs via hydroxylation at C3 followed by dehydratation. However, the lack of apigenin in these incubations ruled out cis dihydrokaempferol as a free intermediate in FNS I catalysis. Furthermore, neither (+)-trans-dihydrokaempferol nor unnatural (-)-trans-dihydrokaempferol and 2 hydroxynaringenin served as a substrate for FNS I. Overall, the data suggest that FNS I has evolved uniquely in some Apiaceae as a paraphyletic gene from FHT, irrespective of the fact that FNS I and FLS catalyze equivalent desaturation reactions. PMID- 12782297 TI - A pentatricopeptide repeat-containing gene that promotes the processing of aberrant atp6 RNA of cytoplasmic male-sterile rice. AB - A fertility restorer gene (Rf-1) of [ms-bo] cytoplasmic male sterility (CMS) in rice has been reported to be responsible for the processing of RNA of aberrant atp6 of mitochondria. We have carried out map-based cloning of the Rf-1 gene and found that a 4.7-kb genomic fragment of a restorer line promoted the processing of aberrant atp6 RNA when introduced into a CMS line. The genomic fragment contained a single open reading frame encoding 18 repeats of the 35 amino acid pentatricopeptide repeat (PPR) motif. The cloned PPR gene is a possible candidate of Rf-1. A non-restoring genotype was identified to have deletions within the coding region. PMID- 12782298 TI - Identification and analysis of catalytic TIM barrel domains in seven further glycoside hydrolase families. AB - Fold recognition results allocate catalytic triose phosphate isomerase (TIM) barrels to seven previously unassigned glycoside hydrolase (GH) families, numbers 29, 44, 50, 71, 84, 85 and 89, enabling prediction of catalytic residues. Modelling of GH family 50 suggests that it may be the common evolutionary ancestor of families 42 and 14. TIM barrels now comprise the catalytic domains of more than half of the assigned GH families, and catalyse a much larger variety of GH reactions than any other catalytic domain architecture. Only 327 GH sequences still have no structurally identified catalytic domain. PMID- 12782299 TI - An E2F site in the 5'-promoter region contributes to serum-dependent up regulation of the human proliferating cell nuclear antigen gene. AB - The synthesis of proliferating cell nuclear antigen (PCNA) is strictly regulated during the cell cycle. To investigate the contribution of the promoter region to the up-regulation of human PCNA expression at the onset of S phase, we have examined 17 putative elements with reporter assays in quiescent L-O2 cells and following serum stimulation. The E2F-like sequence 5'-TTCCCCGCAA-3' located at 84 to -75 is required for the serum-induced transactivation. In electrophoretic mobility shift assays, nuclear extracts from asynchronous L-O2 cells exhibit two binding activities toward the -75 E2F oligonucleotide, and the minor band, whose formation could be interfered with by E2F-1 antibody, represents an S phase specific complex. This is the first demonstration of the E2F site in the human PCNA 5' promoter as a serum-responsive element. PMID- 12782300 TI - Kinetics of inhibition of sperm beta-acrosin activity by suramin. AB - Sperm beta-acrosin activity is inhibited by suramin, a polysulfonated naphthylurea compound with therapeutic potential as a combined antifertility agent and microbicide. A kinetic analysis of enzyme inhibition suggests that three and four molecules of suramin bind to one molecule of ram and boar beta acrosins respectively. Surface charge distribution models of boar beta-acrosin based on its crystal structure indicate several positively charged exosites that represent potential 'docking' regions for suramin. It is hypothesised that the spatial arrangement and distance between these exosites determines the capacity of beta-acrosin to bind suramin. PMID- 12782301 TI - Early changes in glucose and phospholipid metabolism following apoptosis induction by IFN-gamma/TNF-alpha in HT-29 cells. AB - The effects of apoptosis induction on glucose and phospholipid metabolite levels in cancer were studied using human colon adenocarcinoma cells (HT-29). Apoptosis was induced by co-incubation with 200 U/ml tumor necrosis factor (TNF)-alpha for 4, 8 or 15 h, after sensitization with 500 U/ml interferon (IFN)-gamma for 7 h. Perchloric acid extracts were analyzed by (1)H and (31)P nuclear magnetic resonance (NMR) spectroscopy. Significantly increased lactate and NTP (all nucleoside 5'-triphosphates) signals were detected 4 h after apoptosis-inducing IFN-gamma/TNF-alpha treatment, but not in cells which were TNF-alpha-treated without IFN-gamma preincubation. Simultaneous lactate and NTP changes, if confirmed in vivo, may serve as early, non-invasive markers of treatment response in some tumors. PMID- 12782302 TI - Neutrophil elastase up-regulates interleukin-8 via toll-like receptor 4. AB - Cystic fibrosis is characterised in the lungs by high levels of neutrophil elastase (NE). NE induces interleukin-8 (IL-8) expression via an IL-1 receptor associated kinase signalling pathway. Here, we show that these events involve the cell surface membrane bound toll-like receptor 4 (TLR4). We demonstrate that human embryonic kidney (HEK)293 cells transfected with a TLR4 cDNA (HEK-TLR4) express TLR4 mRNA and protein and induce IL-8 promoter activity in response to NE. Treatment of both HEK-TLR4 and human bronchial epithelial cells with NE decreases TLR4 protein expression. Furthermore, a TLR4 neutralising antibody abrogates NE-induced IL-8 production, and induces tolerance to a secondary lipopolysaccharide stimulus. These data implicate TLR4 in NE induced IL-8 expression in bronchial epithelium. PMID- 12782303 TI - Metabolic efficiency of liver mitochondria in rats with decreased thermogenesis. AB - We have studied changes in hepatic mitochondrial efficiency induced by 24-h fasting or acclimation at 29 degrees C, two conditions of reduced thermogenesis. Basal and palmitate-induced proton leak, which contribute to mitochondrial efficiency, are not affected after 24-h fasting, when serum free triiodothyronine decreases significantly and serum free fatty acids increase significantly. In rats at 29 degrees C, in which serum free triiodothyronine and fatty acids decrease significantly, basal proton leak increases significantly, while no variation is found in palmitate-induced proton leak. The present results indicate that mitochondrial efficiency in the liver is not related to a physiological decrease in whole body thermogenesis. PMID- 12782304 TI - Skeletal muscle mitochondrial oxidative capacity and uncoupling protein 3 are differently influenced by semistarvation and refeeding. AB - We investigated, in skeletal muscle mitochondria isolated from semistarved and refed rats, the relation between the protein expression of uncoupling protein 3 (UCP3) and mitochondrial oxidative capacity, assessed as state 4 and state 3 respiration rates in presence of substrates that are either non-lipids (glutamate, succinate) or lipids (palmitoyl CoA, palmitoylcarnitine). During semistarvation, when whole-body thermogenesis is diminished, state 3 respiration was lower than in fed controls by about 30% independently of substrate types, while state 4 respiration was lower by 20% only during succinate oxidation, but UCP3 was unaltered. After 5 days of refeeding, when thermogenesis is still diminished, neither state 4, state 3 nor UCP3 were lower than in controls. Refeeding on a high-fat diet, which exacerbates the suppression of thermogenesis, resulted in a two-fold elevation in UCP3 but no change in state 4 or state 3 respiration. These results during semistarvation and refeeding, in line with those previously reported for fasting, are not in support of the hypothesis that UCP3 is a mediator of adaptive thermogenesis pertaining to weight regulation, and underscore the need for caution in interpreting parallel changes in UCP3 and mitochondrial oxidative capacity as the reflection of mitochondrial uncoupling by UCP3. PMID- 12782305 TI - The metabolic role of human ADH3 functioning as ethanol dehydrogenase. AB - Human class III alcohol dehydrogenase (ADH3), also known as glutathione-dependent formaldehyde dehydrogenase, exhibited non-hyperbolic kinetics with ethanol at a near physiological pH 7.5. The S(0.5) and k(cat) were determined to be 3.4+/-0.3 M and 33+/-3 min(-1), and the Hill coefficient (h) 2.21+/-0.09, indicating positive cooperativity. Strikingly, the S(0.5) for ethanol was found to be 5.4 x 10(6)-fold higher than the K(m) for S-(hydroxymethyl)glutathione, a classic substrate for the enzyme, whereas the k(cat) for the former was 41% lower than that for the latter. Isotope effects on enzyme activity suggest that hydride transfer may be rate-limiting in the oxidation of ethanol. Kinetic simulations using the experimentally determined Hill constant suggest that gastric ADH3 may highly effectively contribute to the first-pass metabolism at 0.5-3 M ethanol, an attainable range in the gastric lumen during alcohol consumption. The positive cooperativity mainly accounts for this metabolic role of ADH3. PMID- 12782306 TI - Mitochondrial and cytosolic expression of human peroxiredoxin 5 in Saccharomyces cerevisiae protect yeast cells from oxidative stress induced by paraquat. AB - Human peroxiredoxin 5 is a recently discovered mitochondrial, peroxisomal and cytosolic thioredoxin peroxidase able to reduce hydrogen peroxide and alkyl hydroperoxides. To gain insight into peroxiredoxin 5 antioxidant role in cell protection, we investigated the resistance of yeast cells expressing human peroxiredoxin 5 in mitochondria or in the cytosol against oxidative stress induced by paraquat. The herbicide paraquat is a redox active drug known to generate superoxide anions in mitochondria and the cytosol of yeast and mammalian cells leading to the formation of several reactive oxygen species. Here, we report that mitochondrial and cytosolic human peroxiredoxin 5 protect yeast cells from cytotoxicity and lipid peroxidation induced by paraquat. PMID- 12782307 TI - Apoptosis triggered redistribution of caspase-9 from cytoplasm to mitochondria. AB - Caspase-9 is an apoptosis initiator protease activated as a response to the mitochondrial damage in the cytoplasmic complex apoptosome. By fluorescence labelling of proteins, confocal microscopy and subcellular fractionations we demonstrate that caspase-9 is in the cytoplasm of non-apoptotic pituitary cells. The activation of apoptosis with rotenone triggers the redistribution of caspase 9 to mitochondria. Experiments using the general caspase inhibitor z-VAD.fmk and the specific caspase-9 inhibitor z-LEHD.fmk show that the caspase-9 redistribution is a regulated process and requires the activity of a caspase other than the caspase-9. We propose that this spatial regulation is required to control the activity of caspase-9. PMID- 12782308 TI - Saccharomyces cerevisiae 14-3-3 proteins Bmh1 and Bmh2 participate in the process of catabolite inactivation of maltose permease. AB - In this study we show that Reg1, the regulatory subunit of the Reg1/Glc7 protein phosphatase (PP1) complex, interacts physically with the two yeast members of the 14-3-3 protein family, Bmh1 and Bmh2. By using different fragments of the Reg1 protein we mapped the interaction domain at the N-terminal part of the protein. We also show that Reg1 and yeast 14-3-3 proteins participate actively in the regulation of the glucose-induced degradation of maltose permease (Mal61). PMID- 12782309 TI - An archaeal homing endonuclease I-PogI cleaves at the insertion site of the neighboring intron, which has no nested open reading frame. AB - Homing endonucleases (HEs) of the LAGLIDADG family cleave intron/inteinless cognate DNA at, or near, the insertion site (IS) of their own intron/intein. Here, we describe a notable exception to this rule. Two introns, Pog.S1205 (length 32 bp) and Pog.S1213 (664 bp), whose ISs are 8 bp apart, exist within the 16S rRNA gene of the archaeon Pyrobaculum oguniense. Pog.S1213 harbors a nested open reading frame (ORF) encoding a 22 kDa monomeric protein, I-PogI, which contains two LAGLIDADG motifs and has optimal DNA cleavage activity at 90 degrees C. Intriguingly, I-PogI cleaves the Pog.S1205-less substrate DNA in the presence or absence of Pog.S1213. The cleavage site (CS) of I-PogI does not coincide with the IS of Pog.S1213 but with that of Pog.S1205. Thus, I-PogI activity both promotes the homing of its own intron, Pog.S1213, and guarantees co-conversion of the ORF-less intron Pog.S1205. PMID- 12782310 TI - Cytokine induction of prolactin receptors mediates prolactin inhibition of nitric oxide synthesis in pulmonary fibroblasts. AB - Prolactin (PRL) has been implicated as a modulator of immune function, and some of its actions may be linked to NO synthesis. Because NO acts as a mediator of inflammation, we speculated that an inflammatory milieu could unmask pathways by which PRL could affect NO synthesis. Here, we show that pro-inflammatory cytokines induce the expression of PRL receptors in pulmonary fibroblasts, allowing PRL to inhibit cytokine-induced NO production and the expression of the inducible nitric oxide synthase (iNOS). Inhibition of iNOS expression by PRL correlates with the phosphorylation of STAT-5b (signal transducer and activator of transcription 5b) and the suppression of expression of IRF-1 (interferon regulatory factor 1), a transcription factor for iNOS. These results reveal previously unrecognized mechanisms by which PRL and PRL receptors may play significant modulatory roles during immune-inflammatory processes. PMID- 12782311 TI - Expression of the VRK (vaccinia-related kinase) gene family of p53 regulators in murine hematopoietic development. AB - The vaccinia-related kinase (VRK) proteins are a new group of three Ser-Thr kinases in the human kinome. VRK proteins are upstream regulators of several transcription factors. VRK1 phosphorylates p53 in Thr-18 within the region of binding to mdm2 preventing their interaction. The tissue distribution of three genes is still largely unknown. In the present report the expression of these genes was analyzed during murine hematopoietic development. The three genes are expressed in fetal liver and peripheral blood, with higher levels between days 11.5 and 13.5, a time when there is a massive expansion of liver cells, and thereafter their expression falls significantly. VRK genes are expressed, particularly at mid-gestation, in embryo thymus and spleen, but in adult thymus and spleen their levels are very low. VRK2 is expressed at lower levels than VRK1 and VRK3 in the mouse embryo. VRK genes play a role during embryonic development of hematopoiesis. PMID- 12782312 TI - BMP-2 regulates cardiomyocyte contractility in a phosphatidylinositol 3 kinase dependent manner. AB - Bone morphogenetic protein-2 (BMP-2) regulates development of heart during vertebrate embryogenesis. In vitro BMP-2 induces differentiation of precardiac cells into mature cardiomyocytes by inducing the expression of cardiac-specific genes. However, the role of BMP-2 and its signaling in other cardiac functions have not been studied. We examined the action of phosphatidylinositol (PI) 3 kinase in isolated adult rat cardiomyocytes. Incubation of rat ventricular cardiomyocytes with BMP-2 increased the PI 3 kinase activity. Ly294002, a pharmacological inhibitor of PI 3 kinase, blocked BMP-2-induced PI 3 kinase activity completely. To investigate the contractility of isolated cardiomyocytes, fractional shortening was examined. BMP-2 significantly increased the percent fractional shortening of the cardiomyocytes. Inhibition of PI 3 kinase activity completely abolished this action of BMP-2. These data indicate that PI 3 kinase regulates BMP-2-induced myocyte contractility. To further confirm this observation, we used adenovirus-mediated gene transfer to express a constitutively active myristoylated catalytic subunit of PI 3 kinase in rat cardiomyocytes. Infection of cardiomyocytes with the adenovirus vector increased the expression of constitutively active PI 3 kinase within 24 h. Expression of constitutively active PI 3 kinase significantly increased cardiomyocyte contractility. Together, these data show for the first time that the growth and differentiation factor, BMP-2, stimulates cardiomyocyte contractility. Also we provide the first evidence that BMP-2-induced PI 3 kinase activity regulates this cardiomyocyte function. PMID- 12782313 TI - Palmitate modulates the early steps of insulin signalling pathway in pancreatic islets. AB - Insulin stimulates its own secretion and synthesis by pancreatic beta-cells. Although the exact molecular mechanism involved is unknown, changes in beta-cell insulin signalling have been recognized as a potential link between insulin resistance and its impaired release, as observed in non-insulin-dependent diabetes. However, insulin resistance is also associated with elevated plasma levels of free fatty acids (FFA) that are well known modulators of insulin secretion by pancreatic islets. This information led us to investigate the effect of FFA on insulin receptor signalling in pancreatic islets. Exposure of pancreatic islets to palmitate caused up-regulation of several insulin-induced activities including tyrosine phosphorylation of insulin receptor and pp185. This is the first evidence that short exposure of these cells to 100 microM palmitate activates the early steps of insulin receptor signalling. 2-Bromopalmitate, a carnitine palmitoyl-CoA transferase-1 inhibitor, did not affect the effect of the fatty acid. Cerulenin, an acylation inhibitor, abolished the palmitate effect on protein levels and phosphorylation of insulin receptor. This result supports the proposition that protein acylation may be an important mechanism by which palmitate exerts its modulating effect on the intracellular insulin signalling pathway in rat pancreatic islets. PMID- 12782314 TI - Quaternary structure of alpha-crustacyanin from lobster as seen by small-angle X ray scattering. AB - The structure of alpha-crustacyanin, the blue carotenoprotein of lobster (Homarus gammarus) carapace, has been investigated for the first time using small-angle X ray scattering. In this paper, we have determined the dimensions of this protein composed of eight heterodimeric subunits of beta-crustacyanin. Analysis of the scattering spectra and estimation of the shape of alpha-crustacyanin show that the protein fits into a cylinder with an axial length of 238 A and a radius of 47.5 A, in which the eight beta-crustacyanin molecules are probably arranged in a helical manner. PMID- 12782315 TI - Subsite mapping of human salivary alpha-amylase and the mutant Y151M. AB - This study characterizes the substrate-binding sites of human salivary alpha amylase (HSA) and its Y151M mutant. It describes the first subsite maps, namely, the number of subsites, the position of cleavage sites and apparent subsite energies. The product pattern and cleavage frequencies were determined by high performance liquid chromatography, utilizing a homologous series of chromophore substituted maltooligosaccharides of degree of polymerization 3-10 as model substrates. The binding region of HSA is composed of four glycone and three aglycone-binding sites, while that of Tyr151Met is composed of four glycone and two aglycone-binding sites. The subsite maps show that Y151M has strikingly decreased binding energy at subsite (+2), where the mutation has occurred (-2.6 kJ/mol), compared to the binding energy at subsite (+2) of HSA (-12.0 kJ/mol). PMID- 12782316 TI - Substitutions in the Escherichia coli RNA polymerase sigma70 factor that affect recognition of extended -10 elements at promoters. AB - Previous work has shown that the base sequence of the DNA segment immediately upstream of the -10 hexamer at bacterial promoters (the extended -10 element) can make a significant contribution to promoter strength. Guided by recently published structural information, we used alanine scanning and suppression mutagenesis of Region 2.4 and Region 3.0 of the Escherichia coli RNA polymerase sigma(70) subunit to identify amino acid sidechains that play a role in recognition of this element. Our study shows that changes in these regions of the sigma(70) subunit can affect the recognition of different extended -10 element sequences. PMID- 12782317 TI - Defined subcomplexes of the A1 ATPase from the archaeon Methanosarcina mazei Go1: biochemical properties and redox regulation. AB - The potential A(1) ATPase genes ahaA, ahaB, ahaC, ahaD, ahaE, ahaF, and ahaG from the anaerobic archaeon Methanosarcina mazei Go1 were overexpressed in Escherichia coli DK8 (pTL2). An A(1) complex was purified to apparent homogeneity and shown by Western blot and N-terminal sequence analyses to contain subunits A, B, C, D, and F but to be devoid of subunits E and G. Further removal of subunit C was without effect on ATPase activity. The enzyme was most active at pH 5.2 and required bisulfite and acetate for maximal activity. Kinetic studies confirmed three new inhibitors for A(1) ATPases (diethylstilbestrol and its derivatives hexestrol and dienestrol) and identified redox modulation as a new type of regulation of archaeal A(1) ATPases. PMID- 12782318 TI - The stomach as a site for anthocyanins absorption from food. AB - The ability of anthocyanins to permeate the gastric mucosa can be suggested as a possible explanation of the fast kinetics of plasma appearance of anthocyanins in rats and humans. This paper presents an in vivo experiment aimed to prove the involvement of the stomach in the absorption of grape anthocyanins in rats. The required analytical selectivity and sensitivity was achieved by high-performance liquid chromatography, diode array detection and mass spectrometry. Malvidin 3 glucoside appeared in both portal and systemic plasma after only 6 min. The average concentrations measured in portal and systemic plasma were 0.650+/-0.162 microM and 0.234+/-0.083 microM (mean+/-S.E.M.), respectively. PMID- 12782319 TI - Induction of MafBx and Murf ubiquitin ligase mRNAs in rat skeletal muscle after LPS injection. AB - MafBx and Murf are two new rat E3 ubiquitin ligases induced in muscle atrophy. Our goal was to investigate whether lipopolysaccharide (LPS) injection, a model of muscle catabolism, is associated with increased expression of MafBx and Murf. LPS (750 microg/100 g body weight) induces MafBx and Murf mRNA (respectively, 23 fold and 33-fold after 12 h; P<0.001). A transient induction of tumor necrosis factor-alpha mRNA (21-fold; P<0.001 at 3 h) and a decrease of insulin like growth factor-I mRNA (50%; P<0.001 at 6 h), two potential regulators of the ubiquitin proteasome system were also demonstrated. In summary, MafBx and Murf mRNA are up regulated in response to LPS and might play a role in the muscle proteolysis observed. PMID- 12782320 TI - The MDM2 RING finger is required for cell cycle-dependent regulation of its protein expression. AB - The MDM2 oncoprotein is overexpressed in many human tumors and cancers. MDM2 functions as an E3 ligase for p53 and for itself. MDM2 also interacts with the retinoblastoma protein (RB) and the transcription factor E2F1 to promote cell cycle S-phase entry. Here, we report that MDM2 protein expression is cell cycle regulated, which is dependent on its RING finger domain and requires Lys446. We show that MDM2 protein is stabilized at S phase. In addition, overexpression of MDM2 results in stimulation of E2F activity and accumulation of cells in S phase. These data suggest that ubiquitination of MDM2 is cell cycle-regulated and that MDM2 may play a role in cell cycle progression. PMID- 12782321 TI - In situ photoaffinity labeling of the target protein for lembehyne A, a neuronal differentiation inducer. AB - A C(36) linear acetylene alcohol, lembehyne A (LB-A), induces neuronal differentiation against neuroblastoma cells morphologically and also functionally. The differentiation and cytostatic effect induced by LB-A was specific to neuroblastoma, Neuro 2A cells. To identify the target protein for LB A, a radioactive photoaffinity probe, [(125)I]18-(2'-azido-5'-iodo-benzoyloxy)-LB 18 ([(125)I]azido-LB-18), was synthesized. As a result of in situ labeling experiments against Neuro 2A cells, a protein of M(r) 30 kDa was photolabeled specifically. This labeling was inhibited in the presence of LB-A or the active analogs of LB-A, whereas the inactive analogs showed no inhibitory effect on this labeling. These results suggest that this protein of M(r) 30 kDa is the target protein for LB-A and may play an important role for the neuronal differentiation in neuroblastoma, Neuro 2A cells. PMID- 12782323 TI - Predicted protein-protein interaction sites from local sequence information. AB - Protein-protein interactions are facilitated by a myriad of residue-residue contacts on the interacting proteins. Identifying the site of interaction in the protein is a key for deciphering its functional mechanisms, and is crucial for drug development. Many studies indicate that the compositions of contacting residues are unique. Here, we describe a neural network that identifies protein protein interfaces from sequence. For the most strongly predicted sites (in 34 of 333 proteins), 94% of the predictions were confirmed experimentally. When 70% of our predictions were right, we correctly predicted at least one interaction site in 20% of the complexes (66/333). These results indicate that the prediction of some interaction sites from sequence alone is possible. Incorporating evolutionary and predicted structural information may improve our method. However, even at this early stage, our tool might already assist wet-lab biology. PMID- 12782322 TI - Identification of cell surface determinants in Candida albicans reveals Tsa1p, a protein differentially localized in the cell. AB - To identify cell surface proteins of Candida albicans, the predominant fungal pathogen in humans, we have established an approach using a membrane impermeable biotin derivative in combination with affinity purification. We were able to identify 29 different proteins under two distinct conditions. Among mannoproteins, heat shock proteins and glycolytic enzymes we found thiol-specific antioxidant-like protein 1 (Tsa1p) to be differentially localized depending on the conditions applied. Only in hyphally grown cells Tsa1p was localized to the cell surface whereas in blastospores no surface but mainly nuclear localization was found. This indicates that cell surface expression of at least some proteins is mediated by differential translocation. PMID- 12782324 TI - The C-terminal peptide of thrombospondin-1 stimulates distinct signaling pathways but induces an activation-independent agglutination of platelets and other cells. AB - A peptide from the C-terminal domain of thrombospondin-1 (4N1-1) has been proposed to stimulate platelet aggregation by a novel mechanism involving both an activation-independent agglutination and an activation-dependent, glycoprotein (GP) IIb/IIIa-mediated aggregation which involves GPVI signaling but does not involve CD47. The present study demonstrates that 4N1-1 stimulated a different pattern of signal transduction pathways than the GPVI agonist convulxin. Furthermore, 4N1-1-induced platelet aggregation was activation-independent and not dependent on GPVI or GPIIb/IIIa. Interestingly, 4N1-1 also stimulated activation-independent agglutination of different megakaryocytic and non megakaryocytic cells. 4N1-1-induced cell agglutination but not platelet signaling was inhibited by anti-CD47 antibodies. PMID- 12782325 TI - Pretreatment of indole-3-carbinol augments TRAIL-induced apoptosis in a prostate cancer cell line, LNCaP. AB - Prostate cancer is one of the most common cancers in men and is the second leading cause of cancer-related deaths in the USA. Many anti-tumor agents against prostate cancer cells have been developed, but their unacceptable systemic toxicity to normal tissues frequently limits their usage in clinics. Several previous studies have demonstrated that tumor necrosis factor-related apoptosis inducing ligand (TRAIL) can induce cell death in a variety of transformed cells including prostate cancer cells, but not normal cells. Indole-3-carbinol (I3C), a phytochemical that is produced in fruits and vegetables, may play an important role in the prevention of many types of cancer, including hormone-related ones such as breast and prostate cancer. In this study, we examined the potential sensitizing effects of I3C on TRAIL-mediated apoptosis in a prostate cancer cell line, LNCaP. When LNCaP cells were incubated with I3C (either 30 or 90 microM) for 24 h and then treated with TRAIL (100 ng/ml), enhanced TRAIL-mediated apoptosis was observed. The enhanced apoptosis measured by poly(ADP-ribose) polymerase and caspase 3 cleavage. We also observed that loss of cell viability after treatment with I3C/TRAIL is greater compared with I3C and TRAIL alone. To determine the molecular mechanisms involved in the enhanced apoptosis, we examined the expression of two TRAIL death receptors (DR4 and DR5) and two TRAIL decoy receptors (DcR1 and DcR2). We found that treatment with I3C induced DR4 and DR5 expression at both transcriptional and translational levels. These findings suggest that I3C may be an effective sensitizer of TRAIL treatment against TRAIL resistant prostate cancer cell lines such as LNCaP. PMID- 12782326 TI - Anti-angiogenic property of edible berry in a model of hemangioma. AB - Hemangiomas represent a powerful model to study in vivo angiogenesis. Monocyte chemotactic protein 1 (MCP-1) is known to be responsible for recruiting macrophages to sites of infection or inflammation and facilitate angiogenesis. Recently we have demonstrated that edible berry extracts potently suppress inducible vascular endothelial growth factor expression and in vitro angiogenesis. Comparative analysis of several berry extracts led to the observation that wild blueberry and a berry mix were most effective. Our goal was to follow up on our findings with wild blueberry and the berry mix (OptiBerry). The present work rests on our current finding that these two berry powders significantly inhibit inducible MCP-1 expression in endothelioma cells. Therefore, we sought to examine the effects of wild blueberry and berry mix in an in vivo model of experimental angiogenesis. Reporter studies showed that the berry powders significantly inhibited basal MCP-1 transcription and inducible nuclear factor kappaB transcription. Endothelioma cells pre-treated with berry powders showed diminished ability to form hemangioma. Histological analysis demonstrated markedly decreased infiltration of macrophages in hemangioma of treated mice compared to placebo-treated controls. The current results provide the first in vivo evidence substantiating the anti-angiogenic property of edible berries. PMID- 12782327 TI - Identification and characterization of plant agmatine iminohydrolase, the last missing link in polyamine biosynthesis of plants. AB - The cloning, expression and characterization of plant agmatine iminohydrolase (AIH, also known as agmatine deiminase, EC 3.5.3.12) is described. Recombinant AIH of Arabidopsis thaliana forms dimers and catalyzes the specific conversion of agmatine to N-carbamoylputrescine and ammonia. Biochemical data suggested that cysteine side chains are involved in catalysis. However, site-directed mutagenesis of the two highly conserved cysteine residues of AIH showed that these cysteines are important but not essential for activity, arguing against a thioester substrate-enzyme intermediate during catalysis. This work represents the completion of the cloning of the arginine decarboxylase pathway genes of higher plants. PMID- 12782328 TI - Recognition of novel viral sequences that associate with the dynein light chain LC8 identified through a pepscan technique. AB - Recent data from multiple laboratories indicate that upon infection, many different families of viruses hijack the dynein motor machinery and become transported in a retrograde manner towards the cell nucleus. In certain cases, one of the dynein light chains, LC8, is involved in this interaction. Using a library of overlapping dodecapeptides synthesized on a cellulose membrane (pepscan technique) we have analyzed the interaction of the dynein light chain LC8 with 17 polypeptides of viral origin. We demonstrate the strong binding of two herpesvirus polypeptides, the human adenovirus protease, vaccinia virus polymerase, human papillomavirus E4 protein, yam mosaic virus polyprotein, human respiratory syncytial virus attachment glycoprotein, human coxsackievirus capsid protein and the product of the AMV179 gene of an insect poxvirus to LC8. Our data corroborate the manipulation of the dynein macromolecular complex of the cell during viral infection and point towards the light chain LC8 as one of the most frequently used targets of virus manipulation. PMID- 12782329 TI - Monovalent cation dependence and preference of GHKL ATPases and kinases. AB - The GHKL phosphotransferase superfamily, characterized by four sequence motifs that form the ATP-binding site, consists of the ATPase domains of type II DNA topoisomerases, Hsp90, and MutL, and bacterial and mitochondrial protein kinases. In addition to a magnesium ion, which is essential for catalysis, a potassium ion bound adjacent to the triphosphate moiety of ATP in a rat mitochondrial protein kinase, BCK (branched-chain alpha-ketoacid dehydrogenase kinase), has been shown to be indispensable for nucleotide binding and hydrolysis. Using X-ray crystallographic, biochemical, and genetic analyses, we find that the monovalent cation-binding site is conserved in MutL, but both Na(+) and K(+) support the MutL ATPase activity. When Ala100 of MutL is substituted by proline, mimicking the K(+)-binding environment in BCK, the mutant MutL protein becomes exclusively dependent on Na(+) for the ATPase activity. The coordination of this Na(+) ion is identical to that of the K(+) ion in BCK and involves four carbonyl oxygen atoms emanating from the hinges of the ATP lid and a non-bridging oxygen of the bound nucleotide. A similar monovalent cation-binding site is found in DNA gyrase with additional coordination by a serine side chain. The conserved and protein specific monovalent cation-binding site is unique to the GHKL superfamily and probably essential for both ATPase and kinase activity. Dependence on different monovalent cations for catalysis may be exploited for future drug design specifically targeting each individual member of the GHKL superfamily. PMID- 12782330 TI - Growth-associated protein-43 is elevated in the injured rat sciatic nerve after low power laser irradiation. AB - Low power laser irradiation (LPLI) has been used in the treatment of peripheral nerve injury. In this study, we verified its therapeutic effect on neuronal regeneration by finding elevated immunoreactivities (IRs) of growth-associated protein-43 (GAP-43), which is up-regulated during neuronal regeneration. Twenty Sprague-Dawley rats received a standardized crush injury of the sciatic nerve, mimicking the clinical situations accompanying partial axonotmesis. The injured nerve received calculated LPLI therapy immediately after injury and for 4 consecutive days thereafter. The walking movements of the animals were scored using the sciatic functional index (SFI). In the laser treated rats, the SFI level was higher in the laser treated animals at 3-4 weeks while the SFIs of the laser treated and untreated rats reached normal levels at 5 weeks after surgery. In immunocytochemical study, although GAP-43 IRs increased both in the untreated control and the LPLI treated groups after injury, the number of GAP-43 IR nerve fibers was much more increased in the LPLI group than those in the control group. The elevated numbers of GAP-43 IR nerve fibers reached a peak 3 weeks after injury, and then declined in both the untreated control and the LPLI groups at 5 weeks, with no differences in the numbers of GAP-43 IR nerve fibers of the two groups at this stage. This immunocytochemical study using GAP-43 antibody study shows for the first time that LPLI has an effect on the early stages of the nerve recovery process following sciatic nerve injury. PMID- 12782331 TI - Figure-ground activity in primary visual cortex (V1) of the monkey matches the speed of behavioral response. AB - To look at an object its position in the visual scene has to be localized and subsequently appropriate oculo-motor behavior needs to be initiated. This kind of behavior is largely controlled by the cortical executive system, such as the frontal eye field. In this report, we analyzed neural activity in the visual cortex in relation to oculo-motor behavior. We show that in a figure-ground detection task, the strength of late modulated activity in the primary visual cortex correlates with the saccade latency. We propose that this may indicate that the variability of reaction times in the detection of a visual stimulus is reflected in low-level visual areas as well as in high-level areas. PMID- 12782332 TI - Mismatch negativity to pitch change: varied stimulus proportions in controlling effects of neural refractoriness on human auditory event-related brain potentials. AB - Based on a memory-comparison process, changes in the pitch of repetitive sounds are pre-attentively detected, reflected by the mismatch negativity (MMN) event related brain potential (ERP). In such oddball sequences, ERP responses are also affected by differential refractory states of frequency-specific afferent cortical neurons. This contamination of MMN can be controlled using an additional blocked sequence of equiprobable tones. The present study investigated effects of varying the in-sequence probabilities of these control tones from 10 to 45%, respectively. Results showed that an equal distribution of all control-sequence tones is not necessary for efficiently removing neural refractoriness effects on the ERPs. The genuine memory-comparison-based frequency-change MMN can be estimated using sequences, within which the number of equiprobable control tones varies between three and nine. PMID- 12782333 TI - Adaptation responses of single avian olfactory bulb neurones. AB - Adaptation in the avian olfactory bulb (OB) was examined by recording the activity of single OB neurones in the chicken (Gallus domesticus) during prolonged (120 s) odour exposure (20 ppm ammonia). The activity of each neurone before, during and after odour stimulation was investigated with analysis examining changes in firing rate and pattern. Of the 73 OB neurones recorded, 22 responded to ammonia stimulation with either inhibition or excitation of firing (both 50%). Their responses then either adapted (lessening of the response, 32%) or sensitized (heightening of the response, 68%) during the odour stimulus period. Both inhibited and excited units underwent adaptation or sensitization and their responses were consistent during odour stimulation. These results demonstrate the role of OB neurones in adaptive responses of the avian olfactory system during prolonged odour stimulation. PMID- 12782334 TI - Heat shock protein-90-induced microglial clearance of exogenous amyloid-beta1-42 in rat hippocampus in vivo. AB - Alzheimer's disease is characterized by the accumulation of extracellular amyloid beta (A beta) fibrils with microglia. In an in vitro microglial culture, we recently found that heat-shock protein-90 (Hsp90) enhanced the microglial phagocytosis and clearance of A beta (1-42) (A beta 42). In this study, we examined the microinjection of A beta 42 in the presence or absence of Hsp90 into the rat hippocampus in vivo. Intrahippocampal injection of A beta 42 alone induced microglial accumulation, and the amount of A beta 42 then gradually decreased. In addition, simultaneous injection with Hsp90 significantly reduced the amount of A beta 42 and increased the production of cytokines. These results suggest that Hsp90 may facilitate microglial A beta 42 clearance in rat brain in vivo. PMID- 12782335 TI - Perfusate oxygen and carbon dioxide concentration influence basal microdialysate levels of striatal glucose and lactate in conscious rats. AB - O(2) concentration ([O(2)]) in air equilibrated solutions at room temperature is three fold higher than that in brain extracellular fluid (ECF), and CO(2) concentration ([CO(2)]) is 100 times lower. Using microdialysis the ECF is routinely dialyzed against glucose free isotonic perfusates containing 200 microM O(2) and 10 microM CO(2). In conscious rats, 2 days after probe implantation, decreasing perfusate [O(2)] from 200 to 68 microM (physiologic level) for 60 min, while maintaining a low [CO(2)] (10 microM), increased striatal dialysate glucose and lactate by 12% and 33%, respectively. The same protocol on the third day essentially had no effect on monoamine metabolites. Decreasing [O(2)] concurrent with increasing [CO(2)] to 1.3 mM (physiologic level) for 60 min increased glucose and lactate by 17% and 37%, respectively. This study demonstrates for dialysis studies of glucose and lactate, perfusates that mimic physiologic ECF [O(2)] and [CO(2)] are more appropriate. PMID- 12782336 TI - Two clusters of serum midkine levels in drug-naive patients with schizophrenia. AB - Midkine (MK) is a heparin-binding growth factor implicated in various biological phenomena such as development of the hippocampus and anxiety. We evaluated serum MK levels of drug-naive (n=15) and medicated (n=25) patients with schizophrenia, and age- and sex-matched normal controls (n=38). The patients showed two clusters in the levels. Four drug-naive patients (26.7%) and two medicated patients (8.0%) had abnormally high values, but no controls did, there being a significant difference in the numbers (P=0.003, Fisher's exact test). Furthermore, in other patients, the mean MK levels in drug-naive schizophrenia (0.30+/-0.10 ng/ml) were significantly (P=0.018, Fisher's protected least significant difference test) decreased than those in the controls (0.40+/-0.12 ng/ml). These suggest that there are two clusters of serum MK abnormalities in drug-naive patients with schizophrenia. PMID- 12782337 TI - Simultaneous analysis of five genetic risk factors in Polish patients with Alzheimer's disease. AB - As Alzheimer's disease (AD) is a complex disease, we decided to estimate how previously reported genetic polymorphisms interact to increase the risk for the disease. Five candidate genes were chosen: apolipoprotein E (APOE), alpha 2 macroglobulin, cathepsin D, myeloperoxidase and nitric oxide synthase. Genotyping was performed in 100 cases of late-onset AD and 100 healthy controls. We found a highly significant difference in APOE epsilon 4 distribution between groups (P<0.005). However, no evidence of association for other studied loci was found. Cumulative analysis of five genetic polymorphisms was performed, but it also failed to reveal any synergistic effect of candidate genes greater than that caused by APOE itself. Our results suggest that the APOE epsilon 4 allele is the only known genetic risk factor for late-onset, sporadic AD. PMID- 12782338 TI - The inhibition of long-term potentiation in the rat dentate gyrus by pro inflammatory cytokines is attenuated in the presence of nicotine. AB - Nicotine has previously been shown to affect both long-term potentiation (LTP) and long-term depression and to reverse age-related impairments of LTP in the hippocampus. Levels of proinflammatory cytokines are known to be elevated with age and to inhibit LTP. In the present study we have investigated the effects of three pro-inflammatory cytokines on nicotine-enhanced LTP in the rat hippocampus in vitro. In the presence of nicotine the inhibitory effect of interleukin-1 beta, interleukin-18 and tumour necrosis factor-alpha on LTP was eliminated. Furthermore, significant depotentiation of established LTP could not be obtained in slices treated with nicotine. These experiments demonstrate that nicotine can reverse the inhibitory effects of pro-inflammatory cytokines on LTP. PMID- 12782339 TI - Dexamethasone blocks the refeeding-induced phosphorylation of cAMP response element-binding protein in the rat hypothalamus. AB - We previously reported that dexamethasone pretreatment abolishes the refeeding induced neuronal nitric oxide synthase (nNOS) expression in the rat paraventricular nucleus (PVN). It was reported that nNOS upstream carries cAMP response element (CRE) and nNOS expression is mediated by a CRE-binding protein (CREB)-dependent mechanism. In this study, CREB phosphorylation was co-localized in the nNOS neurons of the rat PVN regardless of feeding conditions. The relative amount of phosphorylated CREB in the hypothalamic tissue lysates increased by 1 h of refeeding following 48 h of food deprivation, and interestingly, this increase was blocked by dexamethasone administration before the food onset. These results suggest that glucocorticoids exert an inhibitory role in CREB phosphorylation directed by nutritional stimuli in the rat hypothalamus, and this inhibition may be related to nNOS gene expression in this brain region. PMID- 12782341 TI - Cyclophosphamide induces dose- and time-dependent elevations in spleen norepinephrine levels of BALB/c mice. AB - Chemotherapeutic drugs may not only kill rapidly dividing cells but may also alter the extracellular environment of surviving cells. We investigated the possibility that cyclophosphamide might alter the noradrenergic environment of the spleen. Male BALB/cByJ mice were administered a single injection of cyclophosphamide (0, 15, 50, or 100 mg/kg). Seventy-two hours after injection animals receiving 50 or 100 but not 15 mg/kg experienced elevated norepinephrine concentrations (pmol/mg) compared to animals given 0 mg/kg. The time course of changes in norepinephrine concentration was investigated 24-216 h after administration of 50 mg/kg cyclophosphamide; norepinephrine took 48 h to elevate, remained elevated for 48-96 h, and returned to vehicle-treated levels by 120 h. Cyclophosphamide in both experiments reduced spleen mass but did not alter total norepinephrine/spleen. These results suggest that low doses of cyclophosphamide can increase the norepinephrine available to influence cell-cell interactions in the spleen. PMID- 12782340 TI - Changes in cytoskeletal and tight junctional proteins correlate with decreased permeability induced by dexamethasone in cultured rat brain endothelial cells. AB - The blood-brain barrier (BBB) plays an important role in controlling the passage of molecules from the blood to the extracellular fluid environment of the brain. An immortalised rat brain endothelial cell line (GPNT) was used to investigate the mechanisms underlying dexamethasone-induced decrease in paracellular permeability. Following treatment with 1 microM dexamethasone there was a decrease in transmonolayer paracellular permeability mainly to sucrose, fluorescein and dextrans of up to 20 KDa. According to pore theory, these differences in permeability were consistent with a decrease in the number of pores between brain endothelial cells. This effect was accompanied by a concentration of filamentous actin and cortactin to the cell periphery. Concomitantly, the continuity of the tight junctional protein ZO-1 at the cell borders was improved and was associated with an increase in both ZO-1 and occludin expression. By contrast, the expression and distribution of adherens junctional proteins such as beta-catenin and p100/p120 remained unchanged. These observations suggest that glucocorticoids induce a more differentiated BBB phenotype in cultured brain endothelial cells through modification of tight junction structure. PMID- 12782342 TI - Vestibular decompensation in labyrinthectomized rats placed in weightlessness during parabolic flight. AB - The purpose of this study was to determine whether the absence of gravitational cues during weightlessness could alter the posture and static eye deviation of Earth compensated rats with peripheral vestibular lesions. The responses of bilaterally (BL) and unilaterally (UL) labyrinthectomized rats at a compensated stage (40-43 days after lesion) during parabolic flight were compared with those at an acute stage (2-7 h after lesion) on Earth. When free-floating in 0 g, UL animals showed the same postural pattern as during water immersion just after surgery. The most striking observation was a continuous roll body motion at about 4 Hz, and a skewed asymmetric posture. When restrained in 0 g, static eye deviation was also comparable to that observed at an acute stage. A return to a compensated posture and gaze was observed within a few seconds following the end of the weightlessness conditions. BL animals were less affected. These results suggest that vestibular compensation after unilateral lesion can be disrupted momentarily and is a fragile state during which the otolith system in the remaining vestibular apparatus presumably plays a continuous role. PMID- 12782343 TI - Multilocal magnetic resonance perfusion mapping comparing the cerebral hemodynamic effects of decompressive craniectomy versus reperfusion in experimental acute hemispheric stroke in rats. AB - This study examined the hemodynamic effects of craniectomy compared to reperfusion on the temporal evolution of cerebral perfusion in different brain regions in a rat model of focal cerebral ischemia. Three groups were investigated: no treatment, reperfusion or craniectomy at 1 h. Perfusion-weighted magnetic resonance imaging (PWI) was performed serially from 0.5 to 6 h. Relative regional cerebral blood flow was calculated for different regions and infarct volume was assessed by histology at 24 h. As conclusion, both, craniectomy and reperfusion increased cerebral perfusion in the acute phase of cerebral ischemia. While reperfusion resulted in a homogeneous improvement of perfusion in the cortex and basal ganglia, craniectomy improved only cortical perfusion in areas directly under the craniectomy site. PWI is well suited to non-invasively monitor perfusion alterations after aggressive therapeutical approaches in stroke. PMID- 12782345 TI - Tumor necrosis factor alpha polymorphism C-850T is not associated with Alzheimer's disease and vascular dementia in an Italian population. AB - A pathogenic role of inflammatory factors has been proposed both in Alzheimer's disease (AD) and vascular dementia (VD). A previous report indicated the presence of polymorphism C-850T of tumor necrosis factor (TNF) alpha as a genetic risk factor for VD and, associated with apolipoprotein E epsilon 4, for AD. We have assessed the association between TNF-alpha polymorphism and dementias in Italian populations of AD, VD and elderly controls. The influence of TNF-alpha polymorphism on dementia has not been confirmed in this segment of the Italian population. PMID- 12782344 TI - Variation at the ADAM10 gene locus is not associated with Creutzfeldt-Jakob disease. AB - Prion diseases are characterized by the accumulation in the brain of a misfolded and protease-resistant form of the prion protein (PrP(c)). PrP(c) contains an amyloidogenic, neurotoxic sequence that is essential for conversion into PrP(Sc), the pathological isoform. During normal processing, PrP(c) is cleaved at a site within this sequence, and this cleavage is thought to destroy the amyloidogenic potential of the protein. ADAM10, a disintegrin and metalloprotease that plays a key role in the pathogenesis of Alzheimer's disease, was recently shown to use PrP(c) as a substrate. We investigated whether variability in the ADAM10 gene could contribute to the pathogenesis of Creutzfeldt-Jakob disease (CJD), by analyzing a single nucleotide polymorphism (SNP) within ADAM10, as a genetic marker potentially in linkage disequilibrium with a functional polymorphism, in patients with sporadic or variant CJD. We observed no significant differences in ADAM10 genotype or allele frequencies between CJD patients and healthy individuals. Moreover, the distribution of ADAM10 SNP genotypes and alleles did not differ between groups of patients based on genotype at the polymorphic codon 129 of the prion protein gene--the sole major genetic risk factor for CJD identified to date. Our data indicate that ADAM10 is unlikely to confer genetic susceptibility to CJD. PMID- 12782346 TI - Gender and age related expression of Oct-6--a POU III domain transcription factor, in the adult mouse brain. AB - Oct-6 is a POU III domain transcription factor whose primary role is thought to be developmental. It is expressed in embryonic stem cells, Schwann cells, and in neuronal subpopulations during telencephalic development. Its best characterised role is in Schwann cells where it is thought to regulate myelin specific gene expression. Expression of Oct-6 was recently discovered in neurons in post-mortem human schizophrenic specimens while being undetectable in matched controls. This study of human tissue contrasted in a number of regards with earlier studies of rodent brain, and questioned what we can consider to be normal adult expression of this gene. In this study, we have investigated Oct-6 expression via in situ hybridisation and Western blot analysis in normal adult female mice of different ages. We show that both RNA and protein levels of Oct-6 expression are highly sustained in the adult and aging cerebellum, whereas they are attenuated in the telencephalon by PW30 (postnatal week 30). These observations suggest that Oct-6 expression takes place in a sex and age dependent way. PMID- 12782347 TI - Discharge pattern of the sympathetic vertebral nerve activity in kittens in postnatal ontogenesis. AB - Electrical activity of the sympathetic vertebral nerve (VN) was studied in kittens (0-180 days) using frequency domain analyzes. Rhythmical discharges in the studied nerve were observed from the moment of birth and correlated with cardiac (1.2-3 Hz) and respiratory frequency (0.15-0.7 Hz). Amplitude of discharges was minimal in newborn kittens and increased during ontogenesis. Relative spectral power was maximal at cardiac and respiratory frequencies in animals in all age groups. '10-Hz discharges' (8-12 Hz band) in the VN oscillations appeared only in animals older than 20 days of life. In accordance to their sympathetic nerve discharge, kittens become mature at 2 month of their life. PMID- 12782348 TI - Dynamic material properties of the tectorial membrane: a summary. AB - Dynamic material properties of the tectorial membrane (TM) have been measured at audio frequencies in TMs excised from the apical portions of mouse cochleae. We review, integrate, and interpret recent findings. The mechanical point impedance of the TM in the radial, longitudinal, and transverse directions is viscoelastic and has a frequency dependence of the form 1/(K(j2pif)(alpha)) for 107 days), the modified GFP variants exhibit much lower half-lives (between 70 and 165 min) in M. smegmatis. To investigate their utility in the measurement of mycobacterial gene expression, we cloned the promoter region of a putative amino acid efflux pump gene, lysE (Rv1986), from Mycobacterium tuberculosis together with the divergently transcribed, putative lysR-type regulator gene (Rv1985c) upstream of one of the unstable GFP variants. We found that the expression kinetics of the lysRE-gfp fusion were identical throughout the M. smegmatis growth curve to those measured using a conventional lysRE-xylE reporter fusion, peaking upon entry into stationary phase. In addition, it was established that the tagged GFP variants were also unstable in Mycobacterium bovis BCG. Thus, we have demonstrated that unstable GFP variants are suitable reporter genes for monitoring transient gene expression in fast- and slow-growing mycobacteria. PMID- 12782377 TI - Application of a novel Paenibacillus-specific PCR-DGGE method and sequence analysis to assess the diversity of Paenibacillus spp. in the maize rhizosphere. AB - In this study, a Paenibacillus-specific PCR system, based on the specific primer PAEN515F in combination with bacterial primer R1401, was tested and used to amplify specific fragments of the 16S rRNA gene from rhizosphere DNA. The amplicons were used in a second (semi-nested) PCR for DGGE, in which bacterial primers F968GC and R1401 were used. The resulting products were separated into community fingerprints by DGGE. To assess the reliability of the method, the diversity of Paenibacillus species was evaluated on the basis of DNA extracted directly from the rhizospheres of four different cultivars of maize (Zea mays), i.e. CMS04, CMS11, CMS22 and CMS36, sown in two Brazilian field soils (Cerrado and Varzea). In addition, a clone library was generated from the PCR-generated 16S rDNA fragments, and selected clones were sequenced. The results of the bacterial community analyses showed, at the level of clone libraries, that considerable diversity among Paenibacillus spp. was present. The most dominantly found sequences clustered into 12 groups, each one potentially representing a species complex. Sequences closely affiliated with the P. macerans and P. azotofixans complexes were found in all samples, whereas other sequences were scarcer. Clones affiliated with the latter species complex were most abundant, representing 19% of all clones analysed. The Paenibacillus fingerprints generated via semi-nested PCR followed by DGGE showed a clear distinction between the maize plants grown in Cerrado versus Varzea soils. Thus, soil type, instead of maize cultivar type, was the overriding determinative factor that influenced the community structures of the Paenibacillus communities in the rhizospheres investigated. At a lower level (subcluster), there was a trend for maize cultivars CMS11 and CMS22 on the one hand, and CMS36 and CMS04 on the other hand, to cluster together, indicating that these respective pair of cultivars were similar in their Paenibacillus species composition. This trend was tentatively linked to the growth characteristics of these maize cultivars. These results clearly demonstrated the efficacy of the Paenibacillus-specific PCR-DGGE method in describing Paenibacillus species diversity in rhizosphere soils. PMID- 12782378 TI - Detection of Chlamydia suis from clinical specimens: comparison of PCR, antigen ELISA, and culture. AB - Cell culture is still widely regarded as the gold standard in chlamydial diagnosis despite its well-known limitations in terms of sensitivity. On the other hand, the polymerase chain reaction (PCR) has emerged as a promising alternative because of rapidity and high sensitivity. However, validation of methodologies is required before the issue of standardization can be addressed. In the present study, 109 clinical samples (organ tissue, nasal, and faecal swabs) from pigs experimentally infected with Chlamydia suis were examined by cell culture, nested PCR in the ompA gene region, and two different antigen enzyme-linked immunosorbent assays (ELISAs) in order to compare the diagnostic performance of these methods. Culture and PCR produced the highest proportion of concordant results (kappa coefficient 0.712). Among 99 samples, 34 were positive in both assays, 51 were negative in both assays, 12 culture-negatives were positive in PCR, and only 2 culture-positives were negative in PCR. Thus, the sensitivity and specificity of PCR vs. culture as standard were 94.4% and 81.0%, respectively, whereas the corresponding values for culture vs. PCR as standard were 73.9% and 96.2%, respectively. Both ELISA tests performed considerably weaker. The data underline the potential of PCR as a powerful detection method for chlamydiae. PMID- 12782379 TI - Measurement of chlorite dismutase activities in perchlorate respiring bacteria. AB - Chlorite dismutase (CD) catalyzes the disproportionation of chlorite to chloride (ClO(2)(-)-->Cl(-)+O(2)) and is present in bacteria capable of cell respiration using perchlorate or chlorate. The activity of this enzyme has previously been measured by monitoring oxygen evolution using a Clark-type dissolved oxygen (DO) probe. We demonstrate here, using two other methods to measure CD activity (a chloride-specific electrode and ion chromatography (IC)) via chloride production, that the DO probe method underestimates dismutation rates. Of the three methods, the chloride probe was the easiest to use and did not require extensive sample handling or post-experimental analysis. Using the chloride electrode method, we determined whole cell rate constants (V(max)=64 U/mg DW, K(m)=0.17 mM) for the chlorate-grown suspensions of Dechlorosoma sp. strain KJ. We compared the CD activities of strain KJ at a fixed chlorite concentration (0.6 mM) to four other perchlorate respiring bacteria (PRB), and to one non-PRB (Pseudomonas aeruginosa). Chlorate-grown cultures of the five PRB strains had CD activities ranging from 25 to 50 U/mg of cell dry weight (DW), while aerobically grown cultures of the PRB had much lower CD activities (0.5-4 U/mg DW). To our knowledge, this is the first systematic comparison of the different methods to measure CD activities, and the first comparison of CD activities of different PRBs. PMID- 12782381 TI - Analysis of the microbial functional diversity within water-stressed soil communities by flow cytometric analysis and CTC+ cell sorting. AB - Total and active cell counts within soil samples were determined by culture independent methods using flow cytometry and preparative Nycodenz gradient centrifugation. Whole cells were purified from soil cores and total extractable cell counts assessed by SYBR Green II fluorescence, while active cell counts were determined by 5-cyano-2,3-ditolyl tetrazolium chloride reduction (CTC+ cells). Parallel microcosms, maintained at either field water capacity or subjected to drying, indicated that the total extractable cell count remained between 10(8) and 10(9) g(-1) (dry weight). In contrast, the CTC+ active count fell threefold in dried microcosms (6% of total cell count) when compared to wetted microcosms (18% of total cell count). Specifically, these data highlighted an overall deactivation of microbial biomass during water stress, with 16S rDNA analyses of flow-sorted CTC+ cells demonstrating shifts within the active diversity. Flow cytometry coupled with cell purification techniques represents a significant tool for operationally defining an active and redundant microbial component within soil communities and is demonstrated during water stress. PMID- 12782380 TI - Use of denaturing gradient gel electrophoresis for analysis of the stool microbiota of hospitalized patients. AB - Denaturing gradient gel electrophoresis (DGGE) of PCR-amplified ribosomal RNA gene amplicons was used to study the stool microbiota of hospitalized patients and to examine the effect of antibiotic therapy. For one patient, 16 anaerobic species identified by random cloning and sequencing of PCR-amplified rRNA genes from stool were represented by bands on the DGGE gel. DGGE analysis and similarity index comparisons demonstrated that the anaerobic microbiota of this individual remained stable in the absence of antibiotic therapy, was minimally affected by ciprofloxacin but markedly reduced by clindamycin therapy, and recovery of some organisms was evident within days after discontinuation of clindamycin. DGGE analysis of additional patients demonstrated similar disruptions of the intestinal microbiota associated with antibiotic therapy. The DGGE banding patterns of nine patients showed considerable variability, but several bands were shared among patients. Thus, our findings are consistent with previous studies that utilized culture techniques, and suggest that DGGE is a useful technique for analysis of the stool microbiota of hospitalized patients. PMID- 12782382 TI - A microtiter-plate screening method for biofilm disinfection and removal. AB - A quantitative spectrophotometric method was developed to measure the removal and killing efficacy of antibiofilm agents. Biofilms of Pseudomonas aeruginosa or Staphylococcus epidermidis were grown in 96-well plates, treated with an agent, then stained with either the biomass indicator crystal violet or the respiratory indicator 5-cyano-2,3-ditolyl tetrazolium chloride. This rapid screening method is sensitive enough to elucidate concentration-response relationships as well as differences between species responses to treatments. Using these assays, agents can be ranked by their ability to remove or kill biofilm. PMID- 12782383 TI - Sensitivity of PCR detection of Cyclospora cayetanensis in raspberries, basil, and mesclun lettuce. AB - The sensitivity of a polymerase chain reaction (PCR) method for detection of Cyclospora cayetanensis in raspberries, basil, and mesclun lettuce was evaluated. The assay could detect 40 or fewer oocysts per 100 g of raspberries or basil, but had a detection limit of around 1000 per 100 g in mesclun lettuce. PMID- 12782384 TI - A novel colorimetric method to quantify tannase activity of viable bacteria. AB - A novel colorimetric method to quantify tannase activity of viable tannase producing bacterial strains was developed through application of a visual reading method that was to detect the activity qualitatively. The novel method was sensitive enough to quantify the marginal tannase activity of strains that could not be otherwise measured by conventional spectrophotometric or colorimetric methods. PMID- 12782385 TI - Preservation of yeast cell morphology for scanning electron microscopy using 3.28 micro m IR laser irradiation. AB - As an alternative to conventional fixation procedures for scanning electron microscopy (SEM) analysis, yeast cells (Saccharomyces cerevisiae) were irradiated in ambient air, with an intense 3.28-micro m IR laser pulse. The morphology of the irradiated cells was well preserved, while nonirradiated control cells were severely shriveled. PMID- 12782386 TI - Improved recovery of DNA from polyacrylamide gels after in situ DNA footprinting. AB - Methods used to date for the isolation of DNA from polyacrylamide gels are elution based, time-consuming and with low yield in DNA. This paper describes an improved system employing polyacrylamide gels made of a meltable matrix. The new system was successfully applied to in situ DNA footprinting following gel retardation assays. PMID- 12782387 TI - Distribution of Rho family GTPases in the adult rat hippocampus and cerebellum. AB - Small GTPases are monomeric guanine nucleotide binding proteins of 20-25 kDa mass. Rho GTPases belong to the Ras superfamily of small GTPases. The small GTPases of the Rho family have been shown to participate in the organisation of the actin cytoskeleton and signal transduction pathways leading to gene transcription. Recent evidence suggests that Rho family GTPases may play an important role in synaptic communication in the brain, and particularly in synatic plasticity. In this study the distribution of RhoA, RhoB, RhoG, Cdc42, and Rac1 was investigated in hippocampal and cerebellar tissue of adult rat brain using immunohistochemical techniques. Previous studies suggest that distribution of Rho family mRNA is uniform throughout these structures. Here we provide evidence for differences in expression of these proteins between different regions of the hippocampus, and between the molecular and granular layers in the cerebellum. These differences may prove important with regard to the physiological functions of Rho family GTPases. PMID- 12782389 TI - Delayed, but prolonged increases in astrocytic clusterin (ApoJ) mRNA expression following acute cortical spreading depression in the rat: evidence for a role of clusterin in ischemic tolerance. AB - Clusterin is a sulfated glycoprotein produced by neurons and by resting and activated astrocytes that has several putative functions, including protective responses to brain injury. Cortical spreading depression (CSD) is a powerful yet largely benign stimulus that acutely is capable of providing long-lasting ischemic tolerance. The current study investigated possible alterations in expression of clusterin mRNA in the cerebral cortex of the rat at various times after unilateral CSD. Using semiquantitative in situ hybridization histochemistry, significant increases (30-100%; P< or =0.05) in clusterin mRNA were detected in layers I-III and IV-VI of the ipsilateral cortex at 1, 2, 7 and 14 (layers I-III only) days after CSD. Transcript levels in the ipsilateral cortex were again equivalent to contralateral (control) levels at 28 days after CSD. These molecular anatomical studies also revealed that both neurons and nonneuronal cells (presumed reactive astrocytes) increased their expression of clusterin mRNA following CSD. Notably the time-course of increases in clusterin mRNA after CSD (1-14 days) overlaps that during which CSD reportedly provides neuroprotection against subsequent cerebral ischemia. These findings along with other evidence suggest that increased clusterin production and secretion, particularly by astrocytes, could be neuroprotective-perhaps via one or more of its putative actions that include inhibition of complement activation and cytolysis, effects on chemotaxis and apoptosis, and actions as an anti-stress protein chaperone. PMID- 12782388 TI - Circadian oscillator function in embryonic retina and retinal explant cultures. AB - Retinal circadian oscillators regulate many aspects of retinal function. Investigations of these oscillators and the biochemical cascades that entrain them would be greatly facilitated if experimental paradigms could be identified that permit long-term monitoring of retinal circadian oscillator function in vitro. The purpose of this study was to determine if chicken retinas maintained in explant culture conditions could serve in this capacity. Retinal circadian oscillator function was studied by monitoring iodopsin transcription under cyclic light, constant dark, and following reversal of the light cycle. Rhythms observed in the explant cultures were compared to those observed in retinas of embryos (in ovo) and post-hatch chickens. Robust iodopsin transcript rhythms were observed for up to 9 days in explant cultures maintained under cyclic light. These rhythms persisted for 48 h in constant darkness and the time course for re-entrainment of the rhythm to a reversed light/dark cycle was similar to that observed in post hatch chicken retinas. These results show that circadian oscillators located within the retina play a key role in the regulation of iodopsin transcription in retinal explant cultures and in retinas of post-hatch chickens. Interestingly, our data show that iodopsin transcription in retinas of intact embryos is primarily, if not entirely, driven by light. These results show that the circadian oscillators driving iodopsin transcription in embryonic retinal explant cultures exhibit functional characteristics similar to those found in post-hatch chicken retina, supporting use of this paradigm in further studies of entrainment of these oscillators in retina. PMID- 12782390 TI - Characterization of zebrafish proenkephalin reveals novel opioid sequences. AB - Cloning and molecular characterization of an homologous gene to proenkephalin in a teleost, the zebrafish Danio rerio is presented in this paper. The new zebrafish proenkephalin (zfPENK) encodes a 249 amino acid polypeptide that displays an identity of 40% to mammalian PENKs and which contains the consensus sequences for four Met-enkephalins, one Leu-enkephalin and one Met-enkephalin-Gly Tyr, described for the first time in teleosts. Expression studies indicate that zfPENK is selectively expressed in zebrafish brain. Our findings support the concept that PENK genes might have been highly conserved throughout evolution and that its origins might be placed more than 400 million years ago. Moreover, we present evidence that the heptapeptide Met-enkephalin-Gly-Tyr present in fish might be anterior in evolution to the heptapeptide Met-enkephalin-Arg-Phe present in tetrapods. Also another homologous sequence to proenkephalin in zebrafish genome is presented. This sequence might stand for the third exon of a possible duplicate gene of zfPENK. Our findings not only present new data in relation to the evolution of opioid peptides in vertebrates, but also we present a new heptapeptide with putative differential activity than the other peptides derived from the mammalian proenkephalins. Future research will define the functional role of this new heptapeptide in the mechanism that describes opioid activity. PMID- 12782391 TI - Period gene expression in the suprachiasmatic nucleus of behaviorally decoupled hamsters. AB - Circadian rhythms in locomotor activity in nocturnal animals typically show activity during the night followed by quiescence during the day. In hamsters, this activity pattern can be altered by confining them to novel running wheels for a few hours during the day, thereby inducing them to be active. After several days of induced activity, two bouts of activity occur spontaneously, one during the day, and the other at night. This phenomenon is known as behavioral decoupling. In the present study we examined the effects of behavioral decoupling on the pattern of SCN Per expression. Our results show robust expression of Period genes in the SCN of animals killed during either of the two inactive phases within a 24-h period, and weak expression of those genes in the SCN of animals killed during the two active phases. This contrasts with the circadian rhythm of Per expression typically seen in the SCN of entrained or free running animals. Our data support the idea that behavioral decoupling reorganizes the circadian system in a manner different from that produced by constant light induced splitting and constant dark splitting. PMID- 12782392 TI - Effect of haemorrhage on the expression of neurotransmitter-related genes in rat ventrolateral medulla: a quantitative real-time RT-PCR study. AB - The ventrolateral medulla (VLM) has three functionally defined regions that contain catecholamine-synthesising neurons (rostral C1, caudal C1 and A1 regions). Many neuromessengers can alter cardiovascular functions in the VLM. The aims of this study were, first to validate the utility of real-time RT-PCR SYBR Green assay for quantitation of mRNA expression levels of neuromessengers in small site-specific neuronal populations in the VLM, and second to compare the basal mRNA levels of the adrenaline-synthesizing enzyme phenylethanolamine-N methyltransferase (PNMT), neuropeptide Y (NPY) and preproenkephalin (ENK) in the three regions and third to examine the effects of haemorrhage on the expression of these three genes. Rats were anaesthetised with sodium pentobarbital and divided into three groups: perfused, sham-operated and haemorrhaged. A 15% haemorrhage was carried out on the haemorrhaged group. It was found that there are regional differences in the level of mRNA expression for all the three genes: with, in general, decreases from the rostral to caudal regions of VLM. A 15% haemorrhage significantly induced expression of PNMT in the rostral C1 region and NPY in the caudal C1 and A1 regions but had no effect on ENK at any sites, suggesting a differential regulation on the expression of these three genes in the VLM. Our results also demonstrate that real-time RT-PCR is a sensitive and accurate method for quantitative studies on neurotransmitter gene expressions in restricted brain regions. PMID- 12782393 TI - Quantitative RT-PCR reveals a ubiquitous but preferentially neural expression of the KIS gene in rat and human. AB - KIS is the only known protein kinase that possesses an RNA recognition motif. This original structure indicates a role for KIS in the maturation of RNAs possibly by phosphorylating and regulating the activities of RNA associated factors. Another function of KIS has recently been unravelled--it negatively regulates the cdk inhibitor p27Kip1 and thus promotes cell cycle progression through G1. In order to explore the functional expression of this kinase, we quantified its mRNA in a wide range of rat and human tissues, during development and in tumors. In both species, the highest level of KIS gene expression was in adult neural tissues. Interestingly, within the adult rat brain, KIS mRNA is enriched in several areas including the substantia nigra compacta and nuclei of the brain stem. Furthermore, KIS gene expression increases dramatically during brain development. Altogether our results point to a ubiquitous function for KIS together with a particular implication during neural differentiation or in the function of mature neural cells. No dysregulation of KIS gene expression was detected in human tumors from breast, bladder, prostate, liver and kidney origins. On the other hand, the KIS gene was overexpressed in NF1-associated plexiform neurofibromas and malignant peripheral nerve sheath tumors (MPNSTs) as compared to dermal neurofibroma which suggests a possible implication of KIS in the genesis of NF1-associated tumors. PMID- 12782394 TI - Phosphorylation of the alpha4 subunit of human alpha4beta2 nicotinic receptors: role of cAMP-dependent protein kinase (PKA) and protein kinase C (PKC). AB - This study determined whether the alpha4 subunit of human alpha4beta2 neuronal nicotinic receptors is phosphorylated in situ by cyclic AMP-dependent protein kinase (PKA) or protein kinase C (PKC). To accomplish this, human cloned epithelial cells stably transfected with the human alpha4beta2 nicotinic receptor (SH-EP1-halpha4beta2) were incubated with 32P-orthophosphate to label endogenous ATP stores, and the phosphorylation of alpha4 subunits was determined in the absence or presence of PKA or PKC activation. Autoradiographs and immunoblots indicated that alpha4 subunits immunoprecipitated from a membrane preparation of SH-EP1-halpha4beta2 cells exhibited a single 32P-labeled band corresponding to the alpha4 subunit protein; no signals were associated with untransfected SH-EP1 cells. The alpha4 subunits from SH-EP1-halpha4beta2 cells incubated in the absence of the activators exhibited a basal level of phosphorylation that was decreased in the presence of the PKA inhibitor H-89 (5 microM), but unaltered in the presence of the PKC inhibitor Ro-31-8220 (0.1 microM). Activation of PKA by forskolin (10 microM), dibutyryl-cAMP (1 mM), or Sp-8-Br-cAMP (1 mM) enhanced phosphorylation nearly threefold; the inactive isomer, Rp-8-Br-cAMP (1 mM) had no effect. In addition, the forskolin effect was totally blocked by the PKA inhibitor H-89 (5 microM). Activation of PKC by the phorbol esters PDBu (200 nM) or PMA (200 nM) increased alpha4 subunit phosphorylation approximately twofold, and the PDBu effect was blocked by the selective PKC inhibitor Ro-31-8220 (0.1 microM). These findings indicate that the alpha4 subunit of human alpha4beta2 nicotinic receptors is phosphorylated in situ by PKA and PKC. PMID- 12782395 TI - Increased CRH mRNA levels in the rat amygdala during short-term withdrawal from chronic 'binge' cocaine. AB - There is evidence that suggests that increased corticotropin-releasing hormone (CRH) release in the central nucleus of the amygdala underlies the anxiogenic and stress-like consequences of withdrawal that are common in phenomenology to all drugs of abuse. The present studies were undertaken to determine levels of CRH mRNA in the amygdala, and also in the hypothalamus, frontal cortex and brainstem after short-term (2 days) and intermediate-term (10 days) cocaine withdrawal (with continued saline injections) from chronic (14 days) 'binge' pattern cocaine administration (3 x 15 mg/kg per day at hourly intervals). Confirming our recent finding of an activation of stress responsive hypothalamic-pituitary-adrenal activity during early cocaine withdrawal, there was a significant elevation of plasma corticosterone level after 2-day cocaine withdrawal. There was also a significant elevation of CRH mRNA levels in the amygdala, but not in the hypothalamus, frontal cortex or brainstem after 2-day cocaine withdrawal. A negative correlation between amygdalar CRH mRNA and plasma corticosterone levels was found in the 2-day cocaine withdrawn rats but not in control rats, suggesting that CRH neurons in the amygdala may be differentially responsive to glucocorticoids after chronic cocaine exposure and withdrawal. There were no changes in either plasma corticosterone or amygdalar CRH mRNA levels after 10-day cocaine withdrawal. Our findings of an increase in amygdalar CRH gene expression during early cocaine withdrawal support a potentially important role for amygdalar CRH activity in the anxiogenic and aversive consequences of withdrawal from cocaine during a time when humans are most subject to relapse. PMID- 12782396 TI - DNA damage in brain cells of mice treated with an oxidized form of apomorphine. AB - We investigated whether systemic injection of apomorphine and its oxidation derivative 8-oxo-apomorphine-semiquinone (8-OASQ) could induce DNA damage in mice brain, using the single-cell gel assay. 8-OASQ induced DNA damage in the brains at 1 and 3 h, but not at 24 h after treatment whereas apomorphine induced a slight increase in brain DNA damage frequency at 3 h after treatment, suggesting that both drugs display genotoxic activity in brain tissue. PMID- 12782397 TI - The evidence for GABAB receptor-mediated regulation of acid-base balance: involvement of Na+/H+ exchanger and Na+/HCO3- cotransporter. AB - A comparative analysis of the effects of gamma-aminobutyric acid (GABA) on the expressions of Na+/H+ exchanger 1 (NHE1) and Na+/HCO3- cotransporter (NBC) was investigated in order to extend our understanding of the mechanism of GABA receptor-mediated acid-base balance using a gerbil model. In vigabatrin (VGB, GABA degradation inhibitor) treated gerbils, both NHE1 and NBC immunoreactivities in the hippocampus were significantly elevated, as compared with the controls. Analogous to VGB treatment, baclofen (GABAB receptor agonist) treatment also evoked elevations of both NHE1 and NBC expressions in the hippocampus, whilst their expressions were unaffected by muscimol (GABAA receptor agonist) treatment. Therefore, our findings suggest that GABAB receptor-mediated regulation of NHE1 and NBC expressions may participate in acid-base balance in the gerbil hippocampus. PMID- 12782398 TI - The functional roles of 11 beta-HSD1: vascular tissue, testis and brain. AB - Glucocorticoid hormones bind both glucocorticoid receptor (GR) and mineralocorticoid receptor (MR) exerting a broad spectrum of actions in various tissues. The concentrations of glucocorticoid hormones in the target cells are regulated by 11 beta-hydroxysteroid dehydrogenases, type 1 (11 beta-HSD1) and type 2 (11 beta-HSD2). 11 beta-HSD2 is a unidirectional dehydrogenase, which inactivates biologically active glucocorticoid into inert metabolite, while 11 beta-HSD1 is a bi-directional oxidoreductase, which either inactivates biologically active glucocorticoid or activates inert metabolite into active forms. GRs and MRs are present in various tissues and mediate a broad spectrum of physiological actions. The co-existence of 11 beta-HSD1 with these two types of receptors plays an important role in regulation of glucocorticoid actions. This review examines the roles of 11 beta-HSD1 in vascular tissues, testis, brain and other tissues such as placental, retinal and adipose tissues. PMID- 12782399 TI - Evidence that dehydroepiandrosterone, DHEA, directly inhibits GnRH gene expression in GT1-7 hypothalamic neurons. AB - Dehydroepiandrosterone (DHEA) has been reported to have diverse effects on overall physiology, although its mechanism of action and specific receptor are not yet known. We have used the immortalized, clonal GT1-7 hypothalamic neurons to study DHEA effects on gonadotropin-releasing hormone (GnRH) gene expression. DHEA (10(-4) M) downregulates GnRH transcription by 39, 70 and 83% at 24, 36, and 48 h, respectively, while DHEA-sulphate had no effect. Hydroxyflutamide a specific androgen receptor (AR) antagonist, and cyproterone acetate or trilostane, both inhibitors of 3 beta-hydroxysteroid dehydrogenase/delta 4,5 isomerase, the rate-limiting enzyme for the conversion of DHEA to sex steroids, did not affect the ability of DHEA to downregulate GnRH gene expression. We found that GT1-7 cells did not express aromatase, thereby precluding conversion to estrogen. Analysis of [(14)C] DHEA metabolism by thin layer chromatography indicates that the main metabolites produced are 7 alpha- and 7 beta-hydroxy DHEA, and 7-oxo DHEA, although these steroids were not able to repress GnRH gene expression alone. Cell viability studies indicated that the transcriptional repression observed is not due to GT1-7 cell death. Interestingly, SV40 T-antigen mRNA levels, under the control of 2.3 kb of the rat GnRH gene 5' regulatory region, are also repressed by DHEA. Our studies indicate that DHEA has direct effects on GnRH transcription that appear to be unique from those observed after conversion to other steroidogenic compounds. PMID- 12782400 TI - Gonadotropin releasing hormone (GnRH) and estradiol (E(2)) regulation of cell cycle in gonadotrophs. AB - The number of pituitary cells, their size, hormonal content and release and response to external cues varies between day and night and during the estrus cycle. Previous studies have demonstrated that pituitary cells proliferate rhythmically and that estradiol (E(2)) is a mitogen of alpha T3 cells. We, therefore, studied the effect of gonadotropin releasing hormone (GnRH) and E(2), on the cell cycle in primary cultures of mouse pituitary cells and in the gonadotroph cell line L beta T2. We found that GnRH and E(2) modulate the cell cycle in a time dependent manner and induce proliferation in cultures of mouse pituitary and L beta T2 cells. GnRH induces proliferation in cells isolated in the morning of the estrus day and increases the number of cells in G2 stage when isolated in noon and evening. However, the transition into the G1 stage is enabled only by co-addition of E(2) and GnRH. GnRH stimulates LH release from L beta T2 cells after 2 days via exocytosis while after 4 days in culture, the increase in LH release may be accounted for by the increase in cell number. E(2) enhanced the GnRH response after 2 days, and abolished it after 4 days in culture. Furthermore, E(2) has no effect on LH release and cell number after 2 days in culture, however, after 4 days in culture, E(2) had no effect on the total amount of LH released but inhibited LH release per cell due to increase in cell number. Our results show that GnRH and E(2) function to shorten the cell cycle and regulate the cell number of each stage of the cell cycle. The effect of GnRH and E(2) on the cell cycle is dependent on the circadian time. This mechanism may serve to modulate the size and function of the pituitary cell population and consequently the function of pituitary gonadotrophs regulating the surge of LH release before ovulation. PMID- 12782401 TI - Involvement of thyrotropin-releasing hormone receptor, somatostatin receptor subtype 2 and corticotropin-releasing hormone receptor type 1 in the control of chicken thyrotropin secretion. AB - Thyrotropin or thyroid-stimulating hormone (TSH) secretion in the chicken is controlled by several hypothalamic hormones. It is stimulated by thyrotropin releasing hormone (TRH) and corticotropin-releasing hormone (CRH), whereas somatostatin (SRIH) exerts an inhibitory effect. In order to determine the mechanism by which these hypothalamic hormones modulate chicken TSH release, we examined the cellular localization of TRH receptors (TRH-R), CRH receptors type 1 (CRH-R1) and somatostatin subtype 2 receptors (SSTR2) in the chicken pars distalis by in situ hybridization (ISH), combined with immunological staining of thyrotropes. We show that thyrotropes express TRH-Rs and SSTR2s, allowing a direct action of TRH and SRIH at the level of the thyrotropes. CRH-R1 expression is virtually confined to corticotropes, suggesting that CRH-induced adrenocorticotropin release is the result of a direct stimulation of corticotropes, whereas CRH-stimulated TSH release is not directly mediated by the known chicken CRH-R1. Possibly CRH-induced TSH secretion is mediated by a yet unknown type of CRH-R in the chicken. Alternatively, a pro-opiomelanocortin (POMC)-derived peptide, secreted by the corticotropes following CRH stimulation, could act as an activator of TSH secretion in a paracrine way. PMID- 12782402 TI - Cloning and characterization of estrogen receptor alpha in mummichog, Fundulus heteroclitus. AB - Developmental exposure to 17 beta-estradiol (E(2)) induced the death of embryos and fry, malformations, sex reversal, and incomplete ossification of vertebrae and cranial bones in the cyprinodont fish, the mummichog (Fundulus heteroclitus). To clarify the mechanism by which exogenous estrogens caused these developmental effects, we determined the sequence of an estrogen receptor (ER) coding region, encoded by 620 amino acid residues. This region shared 80% identity to that of ER alpha of medaka (Oryzias latipes). Northern blot analysis showed that two ER alpha mRNAs with 5.5 and 4 kb were expressed in the liver. These mRNAs were strongly induced by E(2) stimulation. The 4 kb mRNA was expressed 8 h after treatment, whereas the 5.5 kb mRNA was not induced until 12 h after E(2) stimulation. Vitellogenin (VTG) was expressed 8 h after E(2) stimulation in the male liver. Receptor binding assays using the protein of F. heteroclitus ER alpha (fh ER alpha) ligand binding domain showed that alkylphenols bind to fh ER alpha with a higher affinity (50 times or more) as compared with the human ER alpha. The present results demonstrate that the fh ER alpha has a sequence very similar to that of medaka, and the mRNA for this receptor was induced by E(2) stimulation, followed subsequently by VTG expression. Furthermore, alkylphenols bind to fh ER alpha more efficiently than to human ER alpha. PMID- 12782403 TI - Activation of insulin-like growth factor II signaling by mutant type p53: physiological implications for potentiation of IGF-II signaling by p53 mutant 249. AB - The aim of the present study was to investigate the intracellular mediators of the third base mutant of codon 249 in p53 gene (p53mt249) mutation that potentiate IGF-II dependent IGF-I receptor (IGF-IR) signaling. p53mt249 enhanced IGF-II dependent IGF-IR signaling in p53 negative Hep3B hepatoma cells which were specifically prevented by IGF-IR antibody, alpha IR3 and lovastin. p53mt249 increased the number of IGF-II binding sites with no change in the affinity of IGF-IR. Enhanced levels of IGF-IR expression and transcription were identified in p53mt249 transfected Hep3B cells. Pre-transfection of cultured hepatoma cells with p53mt249 resulted in a three to fourfold increase in IGF-IR phosphorylation and downstream mediator IRS-I phosphorylation but, enhanced more than 15-fold after IGF-II treatment, which coincides well with the cell growth and thymidine uptake results. Our results showed that p53mt249 modulate IGF-II dependent IGF-IR signaling by upregulating IGF-IR and potentiating IGF-IRs where IGF-IRs became more sensitive on treatment with IGF-II. We concluded that p53mt249 stimulates IGF-II dependent IGF-IR signaling by upregulating the expression of both ligand (IGF-II) and receptor (IGF-IR) through an autocrine and/or paracrine loop and we outline the physiological significance of potentiation of IGF-IR by p53 mutation in the development of hepatocellular carcinoma (HCC). PMID- 12782404 TI - Regulation of expression of thyroid hormone receptor isoforms and coactivators in liver and heart by thyroid hormone. AB - Autoregulation of thyroid hormone (TH) receptors (TRs) is a mechanism whereby a cell can regulate its responsiveness to TH. Nuclear coactivators (NCoAs) modulate TH action and may also be important for regulation of TR expression. We have determined the effect of TH withdrawal and treatment on the expression of different isoforms of TR as well as expression of the NCoAs SRC-1, TIF-2 and SRC 3 using quantitative real time polymerase chain reaction. In order to identify the effect that each TR isoform exerts over the expression of the other, NCoA and TR transcripts were measured in liver and heart tissue from wild type mice or mice with deletion of either TR isoform or SRC-1 genes. In liver, regulation of TR beta1 and TR alpha2 subtype expression is inversely related to TH levels and the regulation of TR beta expression is, in part, controlled by TR alpha. In the heart, the opposite is the case, regulation of TR alpha2 and TR beta1 isoform expression is directly related to TH levels and this regulation is primarily controlled by TR alpha. Although NCoAs are, in general, increased in response to hypothyroidism or in states of TH resistance, SRC-1 specifically does not regulate TR isoform expression. We have demonstrated that TR isoforms and NCoAs are autoregulated transcription factors with tissue specificity. PMID- 12782405 TI - PGE(2) stimulates vascular smooth muscle cell proliferation via the EP2 receptor. AB - Prostaglandins released by injured vascular tissue can modulate smooth muscle cell (SMC) proliferation. The mechanism of action of PGE(2) was investigated with porcine coronary artery SMCs obtained by explant culture. DNA and RNA syntheses exhibited a concentration-dependent increase following treatment of quiescent SMCs with PGE(2), while PGI(2) had no effect. PGE(2) also elevated PCNA expression, bromodeoxyuridine incorporation, and cell number, indicative of a hyperplastic growth response. Furthermore, induction of c-fos expression required activation of both phosphatidylinositol 3-kinase and mitogen-activated protein kinase. Contrary to these data, treatment of proliferating cells with PGE(2) caused a reduction in DNA synthesis. A role for PKA in either growth stimulation or inhibition was excluded. Interestingly, only quiescent SMCs expressed EP2 receptors, and the selective EP2 receptor agonist butaprost confirmed that this receptor was essential for growth stimulation and possibly inhibition. These data suggest that the growth state-dependent actions of PGE(2) on SMC proliferation may be mediated via the EP2 receptor. PMID- 12782406 TI - Characterization of receptor-interacting protein RIP140 in the regulation of SF-1 responsive target genes. AB - Receptor-interacting protein (RIP) 140 interacts with several nuclear receptors, but its function in regulation of nuclear receptor action has been debated. Here we have examined the role of RIP140 in regulation of Steroidogenic factor-1 (SF 1)-dependent transcription. SF-1 interacts with RIP140 through its activation function-2 (AF-2) domain. Several domains of RIP140 interact directly with SF-1, but the carboxyl-terminal region containing 4 of its 9 LXXLL motifs showed the strongest SF-1 interaction. Coexpression of RIP140 and SF-1 in different cell types demonstrated that RIP140 acts as a potent corepressor of transcription from the SF-1 responsive cAMP regulatory sequence 2 (CRS2) element of the CYP17 gene and a variety of SF-1 responsive promoter genes. RIP140 also counteracted the stimulatory action of p160/SRC coactivators. The inhibitory effect of RIP140 was partially reversed by Trichostatin A, suggesting a role of histone deacetylase (HDAC) activity in RIP140-mediated repression of SF-1. Quantitation of endogenous coregulator mRNA levels revealed cell type specific differences that could affect the repressor action by overexpressed RIP140. PMID- 12782407 TI - Islet cell hyperplasia in transgenic mice overexpressing EAT/mcl-1, a bcl-2 related gene. AB - EAT/mcl-1 (EAT), a bcl-2 related anti-apoptotic gene, is up-regulated at the early stage of differentiation of human embryonal carcinoma cells; cells which serve as a model for early embryogenesis. We generated transgenic mice for the human EAT gene driven by the EF1 alpha promoter in order to elucidate its functional role in vivo. Histologically, these mice exhibited hyperplasia of Langerhans islet cells; pancreatic cell regions composed of both insulin- and glucagon-producing cells. Furthermore, Bax and Bag-1 -- possible heterodimeric partners for EAT in the anti-apoptotic process -- were up-regulated in islets isolated from the EAT transgenic mice. The insulin tolerance test exhibited no significant difference between the EAT transgenic mice and non-transgenic mice, indicating that islet cell hyperplasia was not due to insulin resistance. In conclusion, EAT transgenic mice exhibit hyperplasia of pancreatic beta cells. EAT may inhibit apoptosis of beta cells, allowing these cells to circumvent the process of apoptosis until the adult stage. PMID- 12782408 TI - Temporal relationships between FSH receptor, type 1 insulin-like growth factor receptor, and aromatase expression during FSH-induced differentiation of bovine granulosa cells maintained in serum-free culture. AB - A serum-free culture system has been developed in ruminants that allows gonadotrophin-responsive induction of oestradiol (E2) production by non differentiated granulosa cells (GC) from small antral follicles. Critical determinants are dose of FSH and insulin-like growth factor (IGF), and the plating density of the GC. Over the first 16 h of culture when cells remained as a dispersed monolayer, expression declined in FSH receptors (FSHr) (P <0.001), IGF type 1 receptor (IGF-1r) (P <0.08) and p450 arom (CYP19, P <0.001). Characteristic GC clusters formed from 16 h and further enlarged between 24 and 48 h, accompanied by marked increases in FSHr (P <0.01), IGF-1r (P <0.05), and p450 arom (P <0.01) expression, and preceded induction and subsequent peak E2 production, at 96 and 144 h, respectively (P <0.01). In conclusion, isolation and dispersion of GC appears to induce reversion to an immature state resulting in loss of receptor expression. Re-establishment of cell-cell communications in the presence of FSH and IGF results in receptor up-regulation and induction of cellular differentiation. PMID- 12782409 TI - Expression of adenylyl cyclase types III and VI in human hyperfunctioning thyroid nodules. AB - Hyperfunctioning thyroid nodules are characterized by the presence of spontaneous somatic mutations responsible for constitutive activation of the cAMP pathway. However, alterations affecting other elements of the cAMP signaling system may counteract the effects of the mutations. In this study, the expression of the adenylyl cyclase (AC) types III and VI was investigated by Western blot in 18 hyperfunctioning thyroid nodules; in 12 samples, we also assessed the presence of TSH receptor (TSHR) or gsp mutations and levels of AC VI and III mRNA. We found that the expression of nodular AC VI (but not AC III) was significantly lower (85.1% of normal, P=0.014) than the expression of both adenylyl cycles types of perinodular tissue from the same patients. Slightly, but not significant differences were detected in nodules with or without mutations and AC protein levels generally showed correlation with the levels of the transcripts detected by RT-PCR. In addition, AC III and AC VI expression levels within a given nodule were characterized by a significant positive correlation. These findings indicate that a diminished expression of AC type VI may be part of the mechanisms occurring in the hyperfunctioning nodules, independently of the presence of TSHR or gsp mutations, which influence the resulting phenotype. PMID- 12782410 TI - Gonadotropins and inhibins along the development of a luteinized rat ovarian tumor. AB - Luteinized intrasplenic ovarian tumors develop in response to high circulating gonadotropins. The relationship between tumor development, gonadotropins and inhibins was studied. Tumor-bearing animals were sacrificed weekly along the first 6 weeks of development. Inhibins were measured by enzyme-linked immunosorbent assay (ELISA), serum gonadotropins, GH and IGF-1 by RIA. Inhibin subunit mRNAs were determined by Northern blot. Tumor histology was examined. Ovarian grafts grew significantly along development. LH increased ten-fold on week 1; a further significant increment was observed on week 3. FSH peaked on weeks 1 and 2 and fell significantly thereafter. Serum inhibins markedly increased on weeks 3-5. Tumor inhibin A content and mRNA levels for alpha and beta A subunits also increased on week 3. Inverse correlations between inhibins and FSH and direct correlations between inhibins and LH were observed. Tumor inhibin A and IGF-1 contents correlated significantly. Increasing levels of luteinization were observed along tumor development. These luteinized tumors develop mainly in response to LH, since growth continues under FSH inhibition. The active inhibin secretion and the positive correlation between inhibins and LH suggests that LH may be the main driving force behind this production, while growth factors produced by the gonads may also participate in their regulation. PMID- 12782411 TI - Chronic hypoxia activates a local angiotensin-generating system in rat carotid body. AB - Evidence exists for the presence of a functional angiotensin system in the carotid body, which can modulate the excitability of the carotid body chemoreceptors. In the present study, the effect of chronic hypoxia on the expression and localization of the angiotensinogen (AGT) and angiotensin converting enzyme (ACE), the two critical components of an intrinsic angiotensin generating system in the rat carotid body, are investigated by in situ hybridization histochemistry, semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis. In situ hybridization showed that the messenger RNA (mRNA) expression of AGT was localized within the type-I glomus cells of the carotid body, which was subjected to be upregulated under the stress of chronic hypoxia. RT-PCR further confirmed a significant increase in the expression of AGT mRNA by chronic hypoxia. Consistently, Western blot analysis demonstrated that chronic hypoxia could elicit the upregulation of AGT protein in chronically hypoxic carotid bodies when compared with their normoxic controls. On the other hand, there was a slight but significant increase in ACE mRNA expression during chronic hypoxia. This study suggests that chronic hypoxia can activate a local angiotensin-generating system in the carotid body, notably its obligatory component AGT. The activation of such an intrinsic, angiotensin generating system in the carotid body during chronic hypoxia should be important in the modulation of cardiopulmonary adaptation in the hypoxic ventilatory response and the electrolyte as well as water homeostasis. PMID- 12782412 TI - PTP1B antisense-treated mice show regulation of genes involved in lipogenesis in liver and fat. AB - Protein tyrosine phosphatases are important regulators of insulin signal transduction. Our studies have shown that in insulin resistant and diabetic ob/ob and db/db mice, reducing the levels of protein tyrosine phosphatase 1B (PTP1B) protein by treatment with a PTP1B antisense oligonucleotide resulted in improved insulin sensitivity and normalized plasma glucose levels. The mechanism by which PTP1B inhibition improves insulin sensitivity is not fully understood. We have used microarray analysis to compare gene expression changes in adipose tissue, liver and muscle of PTP1B antisense-treated ob/ob mice. Our results show that treatment with PTP1B antisense resulted in the downregulation of genes involved in lipogenesis in both fat and liver, and a downregulation of genes involved in adipocyte differentiation in fat, suggesting that PTP1B antisense acts through a different mechanism than thiazolidinedione (TZD) treatment. In summary, microarray results suggest that reduction of PTP1B may alleviate hyperglycemia and enhance insulin sensitivity by a different mechanism than TZD treatment. PMID- 12782413 TI - Pituitary tumorigenesis targeted by the ovine follicle-stimulating hormone beta subunit gene regulatory region in transgenic mice. AB - Targeted tumorigenesis in transgenic mice has been a powerful tool for the study of gene expression and oncogenesis, as well as for the production of differentiated immortal cell lines from rare cell types. Follicle-stimulating hormone (FSH) is secreted by the gonadotrope cells of the anterior pituitary gland and plays a pivotal role in mammalian reproduction. Here we have used the regulatory region of the ovine FSH beta gene to direct expression of the SV40 T antigen oncogene to gonadotrope cells in the pituitary of transgenic mice. Two of five transgenic mouse lines bearing this fusion gene rapidly developed pituitary tumors, with appearance of adenomatous foci as early as 6 weeks of age, resulting in death by 12 weeks of age in both genders. Histologic examination of tumor development over time revealed that increases in cell proliferation and dysplasia were accompanied by decreases in synthesis of pituitary hormones, indicating dedifferentiation of the pituitary cells. Histological features observed in these tumors were in agreement with this rapid transformation of cell phenotype. Tumors were multifocal in origin, and the most highly transformed cell types observed consisted of giant pale basophilic cells with enormous hyperploid nuclei associated with infiltrating neuronal-like cells, which were very abundant at later stages of tumor development. Mitotic indices were much higher in transgenic than wild-type pituitaries, as expected. Morphologic analysis of the gonads of these transgenic mice showed no major developmental differences, as compared to wild-type littermates, however the length of the seminiferous tubules in transgenic males was greater than age-matched wild-type animals. Despite this phenotype difference, both genders were fertile, with normal sperm development observed in males and normal estrous cycle stages in females. Moreover, while 8 - 10-week-old transgenic males had much lower blood levels of FSH than littermates, transgenic female FSH levels were the same as those of wild-type females. These animals offer a unique and potentially useful model of organ specific tumorigenesis, where a multistage pathway of tumor development is evident, both histologically and temporally. Study of such models will advance our knowledge on the physiological and molecular mechanisms involved in gene expression as well as tumor formation. PMID- 12782415 TI - Developmental noise, ageing and cancer. AB - Development is a very robust but far from perfect process, subjected to random variation due to the combined factors that constitute the so-called developmental noise. The effects of early developmental noise may have long-term consequences resulting from slight differences in the make-up and organisation of the former developing system. Here we present evidence suggesting that cancer is not an acquired but an intrinsic process resulting from random factors acting during early development, thus leading to a mixture of susceptibility types that may develop cancer sooner or later, depending on the combination of the environment acting upon such different susceptibility types. We discuss evidence suggesting that some supposedly tumour-suppressor functions, such as those associated with the p53 protein, actually evolved as buffering functions against the early effects of developmental noise that might compromise the stability of embryonic cells and hence of development. Ageing is a stochastic process characterised by progressive failure of somatic maintenance and repair. We put forth the notion that progressive loss of the morphological coherence of the organism (morphological disorder) is a form of ageing, and that morphological disorder is the common theme of most types of cancer. Thus, we suggest that the exhaustion of both developmental constraints and buffering developmental mechanisms link ageing and cancer. Moreover, we propose that cancer may represent one of the most radical forms of ageing, because it generally satisfies the criteria of senescence: intrinsicality, progressiveness and deleteriousness. PMID- 12782414 TI - Localization of activin and inhibin subunits, receptors and SMADs in the mouse mammary gland. AB - Activin and inhibin, two closely related protein hormones, are members of the transforming growth factor beta (TGF beta) superfamily of growth factors. Activin and TGF beta have been associated with mouse mammary gland development and human breast carcinogenesis. TGF beta expression in the mammary gland has been previously described, and was found to be expressed in nonparous tissue and during pregnancy, down-regulated during lactation, and then up-regulated during involution. The expression pattern of activin subunits, receptors and cytoplasmic signaling molecules has not been thoroughly described in post-natal mammary gland development. We hypothesize that activin signaling components are dynamically regulated during mammary gland development, thereby permitting activin to have distinct temporal growth regulatory actions on this tissue. To examine the activin signal transduction system in the mammary gland, tissue from CD1 female mice was dissected from nonparous, lactating day 1, 10, and 20 and post-weaning day 4 animals. The expression of the activin receptors (ActRIIA, ActRIIB and ActRIB), the inhibin co-receptor (betaglycan), and ligand subunit (alpha, beta A and beta B), mRNA was measured by semi-quantitative RT-PCR in these tissues. In addition, the cellular compartmentalization of the activin signaling proteins, including the cytoplasmic signaling co-activators, Smads 2, 3 and 4, were examined by immunohistochemistry. Generally, mRNA abundance of activin signaling components was greatest in the nonparous tissue, and then decreased, whereas protein immunoreactivity for activin signaling components increased during lactation and decreased during involution. The alpha-subunit protein was detected in nonparous and lactating day 1 tissue only. Importantly, Smad 3, but not Smad 2, was detected in epithelial cell nuclei during all time points examined, indicating that activin signaling is mediated by Smad 3 at these times. These findings suggest that activin's growth regulatory role during lactation may be distinguished from that of TGF beta during post-natal mammary development. Future studies will focus on determining the exact role this ligand plays in mammary tissue differentiation and neoplasia. PMID- 12782416 TI - Effect of delta-sleep inducing peptide-containing preparation Deltaran on biomarkers of aging, life span and spontaneous tumor incidence in female SHR mice. AB - From the age of 3 months until their natural deaths, female Swiss-derived SHR mice were subcutaneously injected 5 consecutive days every month with 0.1 ml of normal saline (control) or with 2.5 microg/mouse (approximately 100 microg/kg) of delta-sleep inducing peptide (DSIP, Trp-Ala-Gly-Gly-Asp-Ala-Ser-Gly-Glu) as the preparation Deltaran solved in 0.1 ml of saline. There were 54 mice in each group. The results of this study show that the treatment with Deltaran did not influence food consumption, but decreased the body weight of mice; it slowed down the age-related switching-off of estrous function; it decreased by 22.6% the frequency of chromosome aberrations in bone marrow cells; it did not influence mean life span; and it increased by 17.1% life span of the last 10% of the survivors and by 24.1% maximum life span in comparison with the control group. We also found that treatment with Deltaran significantly decreased total spontaneous tumor incidence (by 2.6-fold), mainly mammary carcinomas and leukemias in mice as compared with the control group. This is the first report on geroprotector and anticarcinogenic effect of DSIP-containing preparation Deltaran. PMID- 12782417 TI - Age-associated changes in SAPK/JNK and p38 MAPK signaling in response to the generation of ROS by 3-nitropropionic acid. AB - Mitochondrial dysfunction has been identified as a major source of oxidative stress in aged tissues. In this study we asked whether activities of components of the SAPK/JNK and p38 MAPK stress response signaling pathways are indicative of oxidative stress in aged mouse livers and whether these pathways are responsive to oxidative stress generated by 3-nitropropionic acid (3-NPA), an inhibitor of complex II (succinic dehydrogenase). We asked whether (a) aging affects the basal activity of the SAPK/JNK stress signaling pathway; (b) specific isoforms of JNK, i.e. 46 or 54 kDa JNKs are activated by 3-NPA; (c) aging affects the response of this signaling pathway to 3-NPA; (d) there is a cross pathway activation of JNK or p38 MAPK by upstream activators. Our studies have shown that although their protein pool levels are not altered, the basal JNK activities using c-Jun as substrate is elevated. Furthermore, in aged livers, JNK activity is induced to a greater extent and takes longer to recover from 3-NPA treatment. The activities of the upstream activators of JNKs, MAP kinase kinase (MKK) 4 and 7, are also elevated in livers of aged C57BL/6 male mice. These activator kinases, which are induced (phosphorylated) by 3-NPA in young livers, are not inducible by this inhibitor in aged livers. In fact, these proteins are highly phosphorylated in the control aged livers and are dephosphorylated in response to 3-NPA. Finally, we demonstrate for the first time that MKK7 serves as an upstream activator of p38 MAPK and that MKK3 and MKK6 activates 54 kDa JNK2 in aged liver. Our studies suggest that failure to respond to 3-NPA may be indicative of the susceptibility of aged tissue to oxidative stress, supporting our hypothesis that aged tissues (especially liver) develop a state of chronic stress even in the absence of a challenge. PMID- 12782418 TI - Proliferation, differentiation and self-renewal of osteoprogenitors in vertebral cell populations from aged and young female rats. AB - A significant contribution to the bone loss associated with aging is likely to be a decline in bone formation. We have characterized and compared the number, capacity for proliferation and differentiation and the self-renewal ability of osteoprogenitors of aged (17-26-month-old) and young (1.5-month-old) female Wistar rats using limiting dilution analyses and continuous subculture experiments. Cells were obtained from outgrowths of explants of lumbar vertebrae (L1-L6) and grown in alpha-minimal essential medium (alpha-MEM), 10% FBS and 50 microg/ml ascorbic acid with or without dexamethasone (Dex; 0.3-300 nM) or progesterone (Prog; 0.01-10 microM). Growth curves for cell populations of both age groups were similar with population doubling times of 27.1 and 26.7 h for the aged and young animals, respectively. Osteoprogenitors from both age groups formed bone nodules when cultured in the presence of either Dex or Prog. Limiting dilution analysis in the presence of 10 nM Dex showed no difference between the aged and young rats in the number of colony forming units-fibroblast (CFU-F), alkaline phosphatase-positive colony forming units-fibroblast (AP+ CFU-F) or colony forming units-osteoblast (CFU-O). No differences were also found for any progenitor within the aged group. Limiting dilution analysis in the presence of 3 microM Prog showed no differences in the numbers of CFU-F, AP+ CFU-F or CFU-O between the aged and young groups or within the aged group. Continuous subculture of cells in the presence of 10 nM Dex revealed that the number of nodules per 10(4) plated cells increased in second subculture over first subculture cells in the young group but decreased in the aged group. Also, in third to fifth subculture cells, the number of nodules was lower in the aged group than in the young group. A similar pattern was observed in the presence of 3 microM Prog. Results indicate that the cell population doubling times, growth characteristics, and the number of CFU-F and osteoprogenitors in vertebral bone cell populations from aged rats and young rats are similar. This suggests that the bone loss associated with aging is not caused by a decrease in osteoprogenitor cell number. However, cell populations from the aged rats showed a reduced capacity for self renewal in vitro, which would ultimately translate into a reduced number of osteoblasts and might be partly responsible for a decrease in bone formation in aged animals. PMID- 12782419 TI - Equine telomeres and telomerase in cellular immortalisation and ageing. AB - To determine the role of telomeres in cellular ageing in equids, we analysed telomere lengths in peripheral blood derived DNA samples from a panel of donkeys (Equus asinus) ranging from 2 to 30 years of age. The average telomere lengths ranged from 7 to 21 kbp and a statistically significant inverse correlation between telomere lengths and donor age was demonstrated. Similarly, telomere lengths in primary fibroblasts isolated from a horse (Equus equus) demonstrated telomeric loss with in vitro ageing when cultured to senescence. We extended this study to evaluate activity of the enzyme telomerase in various equine cell cultures, normal equine tissues and equine benign tumour samples. Initially a panel of equine immortalised and primary cell cultures were evaluated for telomerase activity using a standard telomere repeat amplification protocol (TRAP) assay. High levels of telomerase activity were detected in equine immortalised cells with no activity evident in primary cell cultures. Similarly, no telomerase activity could be detected in normal equine tissues or equine benign tumour samples of the sarcoid or papilloma type. We conclude that telomere attrition may contribute to ageing in equids. However, it would appear that telomerase does not play a major role in the development of the most common benign tumours of the horse. PMID- 12782420 TI - Longevity-associated mitochondrial DNA 5178 A/C polymorphism influences effects of cigarette smoking on serum protein fraction levels in Japanese men. AB - Mitochondrial DNA 5178 adenine/cytosine (mt5178 A/C) polymorphism is reportedly associated with longevity and susceptibility to age-related diseases in Japanese individuals. We previously reported an association between mt5178 A/C polymorphism and serum protein fraction levels in healthy Japanese women. An association between habitual smoking and serum protein fraction levels has also been reported previously. The aim of this study was to examine whether mt5178 A/C polymorphism influenced the effects of habitual smoking on serum protein fraction levels in 321 healthy Japanese men. In mt5178C genotype men, alpha-1 and alpha-2 globulin levels were higher in smokers than in nonsmokers (P<0.001, and P=0.002, respectively). The influence of smoking on these globulin levels depended on cigarette consumption. However, in mt5178A genotype men, no significant difference was observed in alpha-1 or alpha-2 globulin levels between smokers and nonsmokers. These results suggest that longevity-associated mt5178 A/C polymorphism may influence the effects of cigarette smoking on serum protein fraction levels in healthy Japanese men. PMID- 12782421 TI - Developmental outcome at 18 and 24 months of age in very preterm children: a cohort study from 1996 to 1997. AB - OBJECTIVE: To determine the effect of prematurity (gestational age (GA) < 32 weeks) on developmental outcome at the corrected age of 18 and 24 months in a regionally defined, prospective cohort study. STUDY DESIGN: The Leiden Follow-Up Project on Prematurity (LFUPP) includes all live-born infants < 32 weeks GA, born in 1996/1997 in three Dutch health regions (n=266). Mental and psychomotor developmental indices (MDI, PDI) were determined with the Bayley Scales of Infant Development I: > or = -1 S.D.: normal, -2 to -1 S.D.: moderate delay and < -2 S.D.: severe delay. RESULTS: At 18 months 168 (71%) and at 24 months, 151 children (64%) of 235 survivors were assessed. Moderate to severely delayed mental and/or psychomotor development occurred in 40% of the children at both ages. Children lost to follow-up were of lower socioeconomic status and more frequently of non-Dutch origin. Since non-Dutch origin negatively affected the outcome at both test ages, availability of the data of these children would probably have worsened the outcome. Postnatal treatment with dexamethasone was associated with an increased risk of delayed development. Other independent predictors of delayed development were bronchopulmonary dysplasia at 18 months and ethnicity, maternal age at birth, birthweight and gender at 24 months. After adjustment for these other predictors of delayed development, the mean PDI of dexamethasone-treated infants was 16.1 points lower than of non-treated infants at 18 months (p=0.03) and 12.7 points lower at 24 months (p=0.04). CONCLUSIONS: At 18 and 24 months corrected age, 40% of the very prematurely born children had both delayed mental and/or psychomotor development. Treatment with dexamethasone postnatally was a major risk factor for delayed (psychomotor) development. PMID- 12782422 TI - Stability of serial assessments of motor and communication abilities in typically developing infants--implications for screening. AB - BACKGROUND: Neuromaturational theory and dynamic systems theory make different assumptions about the rate of development of motor and communication skills. AIMS: The stability of fine motor, gross motor, and communication scores of infants was evaluated to test these assumptions. STUDY DESIGN: This longitudinal descriptive study evaluated infants in their homes at 9, 11, 13, 16, and 21 months of age. PARTICIPANTS: One hundred and two Canadian children of English speaking parents classified as typically developing at 23 months of age using the Diagnostic Inventory for Screening Children Preschool Screen were included. OUTCOME MEASURES: The Peabody Developmental Motor Scales and the Communication Symbolic Behavior Scales-Developmental Profile were used to assess development at each time. Scores were stable if the 95% confidence intervals around the scores overlapped across all assessments. Correlations evaluated the relationship of scores in a domain over time (normative stability) as well as the relationship of scores between domains (ipsative stability). RESULTS: There was large variability in scores within an infant, among infants and across developmental domains. Typical development is nonlinear rather than occurring at a constant rate. Fine motor, gross motor, and communication skills appear to develop independently. CONCLUSIONS: These results have implications both for developmental screening policies and for early intervention programs. Screening should include multiple domains and multiple time points before referrals are made to early intervention programs. PMID- 12782423 TI - Effects of maternal anxiety on perception of fetal movements in late pregnancy. AB - BACKGROUND: To determine whether maternal state and trait anxiety levels affect maternal perception of fetal movements in the third trimester. METHOD: Forty healthy pregnant nulliparous women not on medication and with a singleton, uncomplicated pregnancy were studied. Maternal anxiety was assessed using the Spielberger State-Trait Anxiety Inventory (STAI, Form-Y) at 36 gestational weeks. At term (37- 40 gestational weeks), fetal movements were recorded simultaneously by the mother and by an ultrasound observer. The ultrasound transducer was positioned to include a cross-section of the fetal trunk and of at least one fetal limb. The ultrasound observer pressed a push-button for as long as she saw a fetal movement on the ultrasound screen. The screen was placed out of sight of the pregnant woman, who pressed another push-button for as long as she felt a fetal movement. The signals were fed into two different channels of a cardiotocograph. Fetal heart rate (FHR) was recorded throughout the examination and classified as FHR patterns A, B, C, or D. Examination time was 120 min. Agreement between maternally recorded and sonographically recorded fetal movements was determined manually for each FHR pattern and was compared between women with low and high state and trait anxiety. The low state anxiety group had STAI scores ranging from 20 to 31, and the high state anxiety group had STAI scores from 32 to 57. The corresponding values for the low and high trait anxiety groups were 20 to 29 and 30 to 54. RESULTS: The agreement between maternally perceived and ultrasonographically recorded fetal movements did not differ between women with low and high state and trait anxiety in any FHR pattern. CONCLUSION: This study does not support that maternal anxiety--within the normal range--affects perception of fetal movements in late pregnancy. PMID- 12782424 TI - Cardiac adaptation in small for gestational age neonates after prenatal hemodynamic disturbances. AB - BACKGROUND: Small for gestational age neonates with prenatal hemodynamic disturbances are at increased risk for neonatal morbidity. Investigations of fetal cardiac function have proved some functional impairments. The aim of the study was to investigate postnatal cardiac adaptation in these neonates in comparison with neonates without prenatal hemodynamic impairments. METHODS AND RESULTS: Forty-one neonates with prenatal hemodynamic disturbances and 40 neonates with undisturbed prenatal hemodynamics were observed during the first 5 days of life. Doppler sonographic measurements of left ventricular time intervals, stroke volume, cardiac output and the incidence of patent ductus arteriosus were obtained in all neonates of both groups. Heart rate and blood pressure were recorded simultaneously. RESULTS: A higher incidence of patent ductus arteriosus and a diminished stroke volume, but increased cardiac output, based on a significantly increased heart rate, were determined in SGA neonates with prenatal hemodynamic disturbances. In contrast, systolic left ventricular time intervals were not changed in these neonates, as expected. CONCLUSIONS: The described findings could be signs of persistent hemodynamic impairments in growth retarded neonates with prenatal disturbed hemodynamics. The neonates revealed a reduced ability to compensate the prenatal hemodynamic disturbances. This aspect should be included in the discussion of perinatal management in cases of severe growth retardation. PMID- 12782426 TI - Energy expenditure and plasma catecholamines in preterm infants with mild chronic lung disease. AB - The present study examined the hypothesis that the energy expenditure (EE) increases during the development of chronic lung disease (CLD) together with serum catecholamines as indicator of stress. Sixteen spontaneously breathing infants with gestational age of 28-34 weeks and birth weight of 870-1920 g were studied. Eight patients were at risk for CLD, eight were healthy controls. Measurements of indirect calorimetry were done weekly at postnatal ages of 2, 3, 4 and 5 weeks. Serum concentrations of adrenaline and noradrenaline were measured by means of a high-pressure liquid chromatography (HPLC) method. The eight CLD risk infants developed mild CLD with FiO(2) of 0.27-0.31 and characteristic radiographic signs at 28 days. Compared to the healthy controls, preterm infants with mild CLD showed increases in EE from week 3 (+67%) to week 5 (+46%). Plasma noradrenaline was increased significantly in the CLD infants when compared to the controls at week 3 (0.7+/-0.3 vs. 0.5+/-0.1 ng/ml; P<0.05) and more pronounced at week 4 (1.4+/-0.2 vs. 0.6+/-0.2 ng/ml; P<0.001) and 5 (1.1+/-0.3 vs. 0.7+/-0.2 ng/ml; P<0.01). Plasma adrenaline was markedly higher in the CLD risk group (mean overall value: 0.64+/-0.1 ng/ml) than in the controls (<0.1 ng/ml in all controls) from week 2 to 5. Regression analysis for the combined values of the infants with and without CLD showed that EE was directly correlated with heart rate, noradrenaline and adrenaline concentration at each of the four study weeks and with respiratory rate at weeks 2 and 3. Increased plasma catecholamine concentrations in preterm infants with CLD suggest that these infants experienced marked stress during the early stages of the disease. Increased EE may in part be a result of this stress. PMID- 12782425 TI - Are there developmentally distinct motor indicators of pain in preterm infants? AB - The aims of this study were to examine preterm infant reactions to pain in detail over prolonged time periods using multiple measures, and to assess the value of including specific body movements of the Neonatal Individualized Developmental Care and Assessment Program (NIDCAP) system to evaluate pain. Ten preterm infants born at 31 weeks mean gestational age (GA) and mean birth weight 1676 g were studied during a routine blood collection in a Level III neonatal intensive care unit (NICU). At 32-week post-conceptional age, computerized physiologic and video recordings were obtained continuously for 60 min (prior to, during and after lance). Motor and facial behaviors were coded independently, using the NIDCAP and the NFCS (Neonatal Facial Coding System), respectively, and compared with heart rate (HR) and oxygen saturation responses. Of the movements hypothesized to be stress cues in the NIDCAP model, extension of arms and legs (80%) and finger splay (70%) were the most common following lance. Contrary to the model, most infants (70%) had lower incidence of twitches and startles post-lance compared to baseline. Whereas all infants showed some NFCS response to lance, for three infants, the magnitude was low. HR increased and oxygen saturation decreased post lance. Infants with more prior pain exposure, lower Apgar, and lower GA at birth, displayed more motor stress cues but less facial activity post-lance. Extension of extremities and finger splay, but not twitches and startles, from the NIDCAP, appear to be stress cues and show promise as clinical pain indicators to supplement facial and physiological pain measures in preterm infants. PMID- 12782427 TI - Ultra-low dose oral naltrexone decreases side effects and potentiates the effect of methadone. PMID- 12782429 TI - Cannabis reduces opioid dose in the treatment of chronic non-cancer pain. PMID- 12782428 TI - Flushing and sweating in an advanced breast cancer patient relieved by olanzapine. PMID- 12782430 TI - Palliative care in a national cancer center: results in 1987 vs. 1993 vs. 2000. AB - In the last few years, palliative care for advanced and terminal cancer patients has undergone considerable evolution. We determined the characteristics of patients admitted to the 4-bed Palliative Care Unit (PCU) of the National Cancer Institute (NCI) of Milan in 1987, 1993 and 2000 to evaluate how our diagnostic and therapeutic approaches have changed over the years. We reviewed the charts of every patient admitted to the PCU in 1987, 1993, and the first ten months of 2000. We recorded demographic data; the primary tumor sites; the main reason for admission; the types of therapies administered (oncologic, analgesic, surgical, neurosurgical analgesic procedures, and supportive therapy); the type and number of cardiological, radiological and endoscopic examinations, as well as specialist consultations; the duration of stay and eventual death on the Unit. There were no significant differences regarding gender, age, primary tumor site and death in hospital of the patients admitted during these years. The time spent in hospital increased over time (P = 0.006). A significant increase was observed in the percentage of patients admitted for supportive therapy (P < 0.001) and investigation concerning the stage of the disease (P < 0.001). There was a significant decrease in admission for invasive analgesic procedures (P < 0.001), as well as for pain diagnosis and/or uncontrolled pain. Uncontrolled pain remained the most frequent reason for admission. Over the years, during hospitalization, 7% to 12% of the patients underwent radiotherapy,1% to 9% had computerized tomography, and 4% to 8% had palliative surgery. More than 50% of the patients received intravenous hydration; a few patients received hypodermoclysis in 1987. Over time, there was a significant increase in "as needed" administration of nonsteroidal anti-inflammatory drugs and a significant reduction in their regular administration (from 24% in 1987 and 1993 to 3% in 2000) (P < 0.001). The use of codeine, tramadol and methadone increased (P < 0.001), whereas the use of oral morphine, buprenorphine and oxycodone decreased in 2000 (P < 0.001). There was a reduction in the use of antidepressants (no significant constant trend) and a significant increase in the use of anticonvulsants, laxatives and pamidronate (P < 0.001). Regularly administered hypnotics decreased in 1993 and increased in 2000 (P < 0.001). Over these years, no significant differences were found in the routes of opioid administration, in route switching and in the mean maximum oral opioid dose (ranging from 108 to 126 mg/day). The percentage of patients undergoing percutaneous cordotomy significantly decreased in 1993 and 2000 (P < 0.001). Over time, there was an increase in requests for specialist consultations, which was significant for neurological, cardiological and oncological consults (P < 0.001). Although the characteristics of the patients admitted to the PCU did not change over these years, there have been significant modifications in our therapeutic approaches, above all in the use of supportive therapy, adjuvant drugs, opioids and neurosurgical invasive procedures. Moreover, a major collaborative interaction with other specialists of the NCI took place with the aim to tailor treatment for each single patient. PMID- 12782431 TI - Effect of recombinant human erythropoietin on quality of life in cancer patients receiving chemotherapy: results of a randomized, controlled trial. AB - The purpose of this study was to assess whether the administration of recombinant human erythropoietin (rHuEPO) would correct anemia and improve the quality of life (QOL) in cancer patients receiving chemotherapy. One hundred twenty-two patients with hemoglobin 20 Kg/m(2), underwent a complete clinical evaluation, including echocardiogram and 24-hour electrocardiogram monitoring to evaluate heart rate variability (HRV) indices. RESULTS: Compared to both thin and control women, patients with AN had greater HRV time domain indices (SDNN: 246.5 +/- 32.4 vs. 170.4 +/- 24 ms and vs. 181 +/- 21.2 ms, p <.001), and in the frequency domain a lower LF/HF ratio (4.2 +/- 1.3 vs. 6.7 +/- 1.2 and vs. 6.8 +/- 1.3 ms, p <.001). AN patients showed reduced left ventricular mass with normal systolic function and typical diastolic patterns, characterized by a lower peak velocity transmitral flow in late diastole (peak A: 35.9 +/- 8.5 vs. 45.2 +/- 7.3 cm/sec and vs. 46.6 +/- 6.3 cm/sec, p <.01), a comparable peak velocity in early diastole (peak E: 92.7 +/- 12.1 vs. 83 +/- 6.2 and vs. 86.8 +/- 9.1 cm/sec, ns) and, subsequently, a greater E/A ratio (2.8 +/- 0.7 vs. 1.8 +/- 0.3 and vs. 1.9 +/- .5, p <.01) than thinness and control groups. Moreover, SDNN was also positively related to E/A ratio (r =.54, p <.01). CONCLUSIONS: Our findings demonstrate a cardiovascular vagal hyperactivity in AN, which appears to influence the ventricular diastolic dynamics. HRV and diastolic function analysis may represent useful tools in monitoring anorexia-induced cardiac modifications. PMID- 12782453 TI - Cardiac response to progressive cycle exercise in moderately obese adolescent females. AB - PURPOSE: To assess cardiac responses to exercise and cardiac functional capacity in moderately obese adolescent females. METHODS: Thirteen healthy females (mean age 13.6 +/- 1.5 years) with a body mass index from 30 to 43 kg per m(2) underwent maximal cycle testing. Cardiac responses were measured by Doppler echocardiogram, and gas exchange variables were determined with open circuit spirometry. Data were analyzed by independent Student's t-tests. RESULTS: Peak oxygen uptake relative to height(3.0) was significantly greater in the obese (570 +/- 90 ml m(-3)) compared to the nonobese controls (485 +/- 60 ml m(-3)). This difference was explained by a higher peak cardiac output (4.50 +/- 1.06 L m(-3) vs. 3.81 +/- 58 L m(-3)) and stroke volume (24 +/- 5 ml m(-3) vs. 19 +/- 3 ml m( 3)) in the obese. Maximal cardiac index was similar in the two groups. There was no evidence of myocardial dysfunction during exercise in either group. CONCLUSIONS: Low aerobic fitness in obese adolescents as indicated by depressed peak VO(2) per kg body mass and limited endurance performance does not reflect decreased cardiac functional capacity. PMID- 12782455 TI - Using social cognitive theory to explain discretionary, "leisure-time" physical exercise among high school students. AB - PURPOSE: To examine whether knowledge of high school students' actions of self regulation, and perceptions of self-efficacy to overcome exercise barriers, social situation, and outcome expectation will predict non-school related moderate and vigorous physical exercise. METHODS: High school students enrolled in introductory Physical Education courses completed questionnaires that targeted selected Social Cognitive Theory variables. They also self-reported their typical "leisure-time" exercise participation using a standardized questionnaire. Bivariate correlation statistic and hierarchical regression were conducted on reports of moderate and vigorous exercise frequency. RESULTS: Each predictor variable was significantly associated with measures of moderate and vigorous exercise frequency. All predictor variables were significant in the final regression model used to explain vigorous exercise. After controlling for the effects of gender, the psychosocial variables explained 29% of variance in vigorous exercise frequency. Three of four predictor variables were significant in the final regression equation used to explain moderate exercise. The final regression equation accounted for 11% of variance in moderate exercise frequency. CONCLUSIONS: Professionals who attempt to increase the prevalence of physical exercise through educational methods should focus on the psychosocial variables utilized in this study. PMID- 12782454 TI - Fat-sugar see-saw in school lunches: impact of a low fat intervention. AB - PURPOSE: To determine the long-term effects of a low fat intervention on sugar content in school lunches METHODS: We calculated contributions of total sugars, 6 specific sugars, and selected nutrients in National School Lunch Program meals served in 56 former intervention, 20 randomly selected control schools that had participated 3 years before in a low fat intervention, and 12 additional schools from neighboring school districts that had never been exposed to the intervention, whose goal was the lowering of total fat to under 30% of calories and saturated fat to under 10% calories. Analysis of variance adjusted for region was used to compare treatment groups. Pearson partial correlations controlling for the effects of region and treatment group were used to assess the strength of sugar and fat relationships. RESULTS: Three years after the low fat intervention, former intervention, control and unexposed schools lunches were similar in mean total sugars (25% of calories), and "added" sugars (e.g. sucrose, glucose, galactose, and maltose) at 15% of calories; and differed only in their lactose content. As percent of calories from fat or saturated fat in lunches decreased, that from sugars increased. Lunches that met reduced saturated fat goals were significantly higher than those not meeting goals in percent of calories from sugars both in meals as offered (27.6 +/- 0.3% vs. 26.2 +/- 0.3 SE p =.004) and as served (26.5 +/- 0.4 vs. 23.9 +/- 0.4 p =.009). Lunches meeting reduced total fat goals were significantly higher only in percent of calories from sugars as served. Seventy-five percent of total sugar in lunches offered was from chocolate milk, fruit/fruit juices, and white milk. More "added" sugar came from high micronutrient foods, such as fruit, fruit juices, and chocolate milk than from desserts and entrees. CONCLUSIONS: The existence of a fat-sugar "see-saw" makes it important to emphasize substitutions of fat and saturated fat with starches and fiber in school lunches. PMID- 12782456 TI - Reproductive health in school-based health centers: findings from the 1998-99 census of school-based health centers. AB - PURPOSE: To describe the state of reproductive health services, including access to contraception and health center policies, among school-based health centers (SBHCs) serving adolescents in the United States METHODS: We examined questionnaire data on provision of reproductive health services from the 1998-99 Census of School-Based Health Centers (response rate 70%). We examined 551 SBHCs in schools with high or middle school grades. We used logistic regression to define factors independently associated with services and policies. RESULTS: Most SBHCs (76%) were open full-time; over one-half (51%) of centers had opened in the past 4 years. Services provided, either on-site or by referral, included gynecological examinations (95%), pregnancy testing (96%), sexually transmitted disease (STD) diagnosis and treatment (95%), Human Immunodeficiency Virus (HIV) counseling (94%), HIV testing (93%), oral contraceptive pills (89%), condoms (88%), Depo-Provera (88%), Norplant (78%), and emergency contraception (77%). Counseling, screening, pregnancy testing, and STD/HIV services were often provided on-site (range 55%-82%); contraception was often provided only by referral (on-site availability = 3%-28%). SBHCs with more provider staffing were more likely to provide services on-site; rural SBHCs and those serving younger grades were less likely to provide these services on-site. Over three-quarters (76%) of SBHCs reported prohibitions about providing contraceptive services on site; the sources of these prohibitions included school district policy (74%), school policy (30%), state law (13%), and health center policy (12%). While SBHCs generally required parental permission for general health services, many allowed adolescents to access care independently for certain services including STD care (48%) and family planning (40%). Older SBHCs were more likely to allow independent access. CONCLUSIONS: SBHCs provide a broad range of reproductive health services directly or via referral; however, they often face institutional and logistical barriers to providing recommended reproductive health care. PMID- 12782457 TI - Second births among teenage mothers: program results and statistical methods. AB - This paper uses survival analysis to examine three large-scale, multi-site, randomized, controlled programs that attempted to prevent or delay second births to teenagers. Statistically significant differences in the hypothesized direction were found between the intervention and the control groups in the Elmira and Memphis Home Visitation sites. No statistically significant differences in the hypothesized direction were found in the Teen Parent Welfare Demonstration overall or in any of its three sites or in all New Chance sites combined. Delaying second pregnancies among teenagers requires intensive efforts. Survival analysis is a more accurate and useful way of presenting program results than simple analysis of the proportion of women with a second birth. PMID- 12782458 TI - Factors associated with self-esteem in pre-adolescents and adolescents with cerebral palsy. AB - Predictors of self-esteem were examined in 50 pre-adolescents and adolescents with cerebral palsy. On average, self-esteem was high, although 30% scored below a cut-point for low self-esteem. Self-esteem was bivariately associated with female gender, better physician-assessed functional ability, greater perception of the impact of the disability, and higher perceived parent overprotectiveness. In a multivariate model, only perceived impact of the disability remained significant. PMID- 12782459 TI - Determination of D- and L-alanine concentrations using a pyruvic acid sensor. AB - The concentrations of D- and L-alanine in bivalves are useful as indicators of environmental pollution. Amino acid oxidase with a low substrate specificity catalyzes the oxidation of various amino acids. Among the various amino acids, pyruvic acid can be generated from alanine only by the catalytic oxidative reaction of this oxidase. Therefore, in this study, the concentrations of D- and L-alanine were determined from the concentration of pyruvic acid, which was determined from the consumption of oxygen based on the oxidative reaction of pyruvate oxidase. From this point of view, there is a very strong possibility that biosensors utilizing enzymes with a low substrate specificity can be developed. The results obtained were as follows. (1) The optimum conditions for the use of pyruvic acid sensor were as follows: temperature of 25 degrees C, pH of 6.8, flow rate of 0.1 ml/min, thiamin diphosphate concentration of 1.5 mM, and injection volume of 50 microl. (2) D-Alanine and L-alanine optimally reacted with D- and L-amino acid oxidase at 30 degrees C, pH 8.2, for 30 min and at 37 degrees C, pH 7.8, for 90 min, respectively. (3) The linear relationships between the concentrations of D- and L-alanine and the output of the sensor were obtained at 3.56-106.8 microg of D-alanine and 5.34-71.3 microg of L-alanine. (4) The concentrations of D- and L-alanine in Meretrix iusoria, Patinopecten yessonsi, and Corbicula leana obtained by the proposed assay were in good agreement with those determined by a conventional method. PMID- 12782460 TI - A single-channel sensor based on gramicidin controlled by molecular recognition at bilayer lipid membranes containing receptor. AB - A novel ion-channel sensor based on a membrane bound receptor and a single gramicidin channel is described, in which the binding of an analyte to the membrane bound receptor modulates the single-channel activity of gramicidin. The sensor is composed of a planar bilayer lipid membrane (BLM) containing biotin labeled phosphatidylethanolamine as receptor for avidin and gramicidin as signal transducer. When the receptor catches an analyte (avidin or ferritin-labeled avidin (FA)) at the membrane surface, the bilayer structure is locally distorted and the gramicidin monomer/dimer kinetics is modulated in a manner that the fraction of channel opening with a short lifetime ( < or = 100 ms) to the total opening events increases. The fraction was found to increase with the concentration of avidin from 1.0 x 10(-9) to 1.0 x 10(-6) M and of FA from 1.0 x 10(-9) to 1.0 x 10(-8) M. With dinitrophenyl-labeled PE embedded as receptor in the BLM for monoclonal anti-dinitrophenyl antibody (anti-DNP), the fraction of channel openings ( < or = 100 ms) increased with the concentration of anti-DNP from 2.0 x 10(-9) to 2.0 x 10(-7) g/ml. Bovine serum albumin (BSA) and anti-BSA antibody caused no changes in the channel opening. The possible mechanism of analyte-induced modulation of single-channel activity of gramicidin is also discussed. PMID- 12782461 TI - A fractal analysis of protein to DNA binding kinetics using biosensors. AB - A fractal analysis of a confirmative nature only is presented for the binding of estrogen receptor (ER) in solution to its corresponding DNA (estrogen response element, ERE) immobilized on a sensor chip surface [J. Biol. Chem. 272 (1997) 11384], and for the cooperative binding of human 1,25-dihydroxyvitamin D(3) receptor (VDR) to DNA with the 9-cis-retinoic acid receptor (RXR) [Biochemistry 35 (1996) 3309]. Ligands were also used to modulate the first reaction. Data taken from the literature may be modeled by using a single- or a dual-fractal analysis. Relationships are presented for the binding rate coefficient as a function of either the analyte concentration in solution or the fractal dimension that exists on the biosensor surface. The binding rate expressions developed exhibit a wide range of dependence on the degree of heterogeneity that exists on the surface, ranging from sensitive (order of dependence equal to 1.202) to very sensitive (order of dependence equal to 12.239). In general, the binding rate coefficient increases as the degree of heterogeneity or the fractal dimension of the surface increases. The predictive relationships presented provide further physical insights into the reactions occurring on the biosensor surface. Even though these reactions are occurring on the biosensor surface, the relationships presented should assist in understanding and in possibly manipulating the reactions occurring on cellular surfaces. PMID- 12782462 TI - Sol-gel-derived titanium oxide/copolymer composite based glucose biosensor. AB - A new type of sol-gel-derived titanium oxide/copolymer composite material was developed and used for the construction of glucose biosensor. The composite material merged the best properties of the inorganic species, titanium oxide and the organic copolymer, poly(vinyl alcohol) grafting 4-vinylpyridine (PVA-g-PVP). The glucose oxidase entrapped in the composite matrix retained its bioactivity. Morphologies of the composite-modified electrode and the enzyme electrode were characterized with a scanning electron microscope. The dependence of the current responses on enzyme-loading and pH was studied. The response time of the biosensor was < 20 s and the linear range was up to 9 microM with a sensitivity of 405 nA/microM. The biosensor was stable for at least 1 month. In addition, the tetrathiafulvalene-mediated enzyme electrode was constructed for the decrease of detection potential and the effect of three common physiological sources that might interfere was also investigated. PMID- 12782464 TI - A bioluminescent sensor for high throughput toxicity classification. AB - A high throughput toxicity monitoring and classification biosensor system has been successfully developed using four immobilized bioluminescent Escherichia coli strains, DPD2511, DPD2540, DPD2794 and TV1061, which have plasmids bearing a fusion of a specific promoter to the luxCDABE operon. The bioluminescence of DPD2511 increases in the presence of oxidative damage, DPD2540 by membrane damage, DPD2794 by DNA damage and TV1061 by protein damage. In the developed biosensor these strains are immobilized in a single 96 well plate using an LB agar matrix, and are able to detect the toxicities of hydrogen peroxide, phenol and mitomycin C in water samples. As the concentration of each chemical was increased, the bioluminescence levels from the corresponding wells, containing either DPD2511, DPD2540, DPD2794 or TV1061, increased. This increase in bioluminescence followed a dose dependent response to the toxic chemicals within a specific concentration range. In particular, each test requires only 4 h to give clear bioluminescent response signature. Storage of the biosensor at 4 degrees C for 2 weeks caused no change in its dose-dependent response. The fast and easy detection of oxidative, membrane, protein and DNA damaging agents in aqueous environments is possible due to the high throughput capability of this biosensor. PMID- 12782463 TI - Determination of organophosphorus aromatic nitro insecticides and p-nitrophenol by microbial-cell respiratory activity. AB - We examined the possibility of measuring the organophosphorus aromatic nitro insecticides metaphos and sumithion as well as their hydrolysis product p nitrophenol (PNP) by the specific respiratory activity (SRA) of Pseudomonas putida C-11, P. putida BA-11, and Acinetobacter calcoaceticum A-122. The plots of cellular SRA against the two insecticides and PNP were linear over the ranges of 0.5-2.5 microM for P. putida C-11 and BA-11 and 0.5-1.0 microM for A. calcoaceticum A-122. P. putida BA-11 showed the greatest respiratory-response selectivity in the determination of the test substrates. We made comparison studies of the SRA of cells immobilised by two methods: carrier-surface adsorption and inclusion in various gels. We discuss the feasibility of developing a microbial sensor system for the determination of metaphos, sumithion, and PNP in aqueous media. PMID- 12782465 TI - Co-immobilized microbial biosensor for BOD estimation based on sol-gel derived composite material. AB - A novel type of biochemical oxygen demand (BOD) biosensor was developed for water monitor, based on co-immobilizing of Trichosporon cutaneum and Bacillus subtilis in the sol-gel derived composite material which is composed of silica and the grafting copolymer of poly (vinyl alcohol) and 4-vinylpyridine (PVA-g-P(4-VP)). Factors that influence the performance of the resulting biosensor were examined. The biodegradable substrate spectrum could be expanded by the co-immobilized microorganisms. The biosensor prepared also exhibited good reproducibility and long-term stability. Good agreement was obtained between the results of the sensor BOD measurement and those obtained from conventional BOD(5) method for water samples. PMID- 12782466 TI - Electrochemical transducers based on surfactant bilayers for the direct detection of affinity interactions. AB - The simple methods for the preparing of direct affinity sensors are proposed. The proposed method consists of the immobilizations of either oligonucleotide or antibodies as recognizing elements onto the surfactant bilayer. For DNA-sensor we propose to immobilize oligonucleotide by spontaneous infiltration of hydrocarbon chain bound to oligonucleotide pentadecathymidylate (dT(15)) into the hydrophobic region of surfactant bilayer. The adsorption of antibodies on bilayer surface has resulted in immunosensor development. The direct detection of affinity interactions in both cases has been investigated by impedance spectroscopy. At both studies the significant changes in impedance spectra have observed. The dynamics of response manifestation have been followed the specific DNA-coupling causing the decrease of real part of impedance, whereas the antibody-antigen interaction caused the increase of real part. The obtained results are promising for the development of impedimetric affinity sensors for clinical or environmental applications. PMID- 12782467 TI - Investigation of complexation of immobilized metallothionein with Zn(II) and Cd(II) ions using piezoelectric crystals. AB - The aim of this study is to investigate complexation of metallothionein (MT) with cadmium and zinc ions. An oligopeptide (i.e. Lys-Cys-Thr-Cys-Cys-Ala), a fragment of MT was covalently immobilized onto piezoelectric crystals, which were first treated with ethylene diamine plasma in a glow-discharge apparatus, and then were chemically reacted with glutaraldehyde. Complexation of the immobilized MT with Zn(II) and Cd(II) ions in aqueous media was followed by recording the changes of the frequency shifts of the piezoelectric quartz crystals. The amount of Cd(II) ions interacted with the immobilized MT molecules was the highest at pH 7.4, and decreased with an increase in the pH of the medium, in parallel to the decrease in the amount of immobilized MT. The number of Zn(II) ions interacted with the immobilized MT molecules was higher than the number of Cd(II) ions when the adsorption was from solutions containing a single-metal ion with the same ion concentrations. In consecutive adsorption studies, we observed that the type of metal ions used in the first interaction is important. These experiments showed also that there is an exchange between the metal ions, and competition provokes adsorption of both ions due to synergistic-antagonistic effects. PMID- 12782468 TI - Development and optimisation of biosensors based on pH-sensitive field effect transistors and cholinesterases for sensitive detection of solanaceous glycoalkaloids. AB - Highly sensitive biosensors based on pH-sensitive field effect transistors and cholinesterases for detection of solanaceous glycoalkaloids have been developed, characterised and optimised. The main analytical characteristics of the biosensors developed have been studied under different conditions and an optimal experimental protocol for glycoalkaloids determination in model solution has been proposed. Using such a biosensor and an enzyme reversible inhibition effect, the total potato glycoalkaloids content can be determined within the range of 0.2-100 microM depending on the type of alkaloid, with lowest detection limits of 0.2 microM for alpha-chaconine, 0.5 microM for alpha-solanine and 1 microM for solanidine. The dynamic ranges for the compounds examined show that such biosensors are suitable for a quantitative detection of glycoalkaloids in real potato samples. High reproducibility, operational and storage stability of the biosensor developed have been shown. PMID- 12782469 TI - Fluorescence quenching competitive immunoassay in micro droplets. AB - A fluorescence quenching competitive immunoassay in micro droplets was applied to the sensitive detection of the pyrethroid insecticide, esfenvalerate. Laser induced fluorescence from rhodamine dye was used as a marker. The competitive immunoreaction was performed in micro droplets generated by a vibrating orifice aerosol generator system with a 10-microm diameter orifice. Fluorescence that was emitted from the droplets was detected by a 1/8 m imaging spectrograph with a 512 x 512 thermoelectrically cooled, charged-coupled device camera. The conjugate of esfenvalerate with rhodamine exhibited similar fluorescence to that of pure rhodamine 6G. When anti-esfenvalerate antibodies were added to the droplets, the fluorescence decreased. The reduction in emission was due to a strong quenching effect that arises from the interaction between the protein and rhodamine molecules following the antigen-antibody reaction. When a sample of esfenvalerate was added to the droplets, the release of the conjugated rhodamine from the antigen-antibody complex allowed the fluorescence signal to recover. An assay in a picoliter droplet sample was shown to enable detection down to approximately 0.1 nM. A very small mass of analyte could be detected with this method. A sample of river water was used to gauge the impact of matrix effects and was shown to give rise to negligible interference with the immunoassay. PMID- 12782470 TI - Assessing the feasibility of using neural precursor cells and peripheral blood mononuclear cells for detection of bioactive Sindbis virus. AB - Viruses form a significant class of bio-threat agents. Currently, the only method to determine the bioactivity of viruses in vitro is to measure viral and cellular responses after co-incubation of cells with virus. Our goal is to find biomarkers for classification of agents, establishment of bioactivity, and/or prediction of disease outcomes. To begin development of a cell-based biosensor for detection of bioactive Sindbis virus (SV), our model analyte, we surveyed the outcomes of SV interaction with primary rat neural precursor cells (NPC) and human peripheral blood mononuclear cells (PBMC). Confocal fluorescence analysis of NPC treated with recombinant SV carrying green-fluorescent-protein (SV-GFP) showed that most cells were GFP positive by day 1 post inoculation. 4',6-Diamidino-2-phenylindole dihydrochloride (DAPI) staining of the nucleus showed nuclear condensation and fragmentation, and the percentage of TUNEL positive cells were higher in virus treated cells than in mock-treated control. Also, there were less BrdU positive cells in virus-treated cells compared to control. Thus, SV infects NPC, decreases cellular proliferation, and induces cell death via apoptosis. PBMC were treated with SV- or UV-inactivated SV. By day 5 post infection, there were fewer adherent cells in SV-treated PBMC compared to UV-inactivated SV treated PBMC. However, the percentage of viable cells remained the same, and virus growth curves showed only clearance of virus. Thus, SV induces detachment of a subpopulation of PBMC while not killing most of the cells. Together, these results indicate that NPC and PBMC respond to bioactive SV inoculation, suggesting potential use as detectors of SV in cell-based biosensor paradigm. These studies also provide the rationale, time scale, and phenotypic correlates for further studies with gene expression arrays. PMID- 12782471 TI - Amperometric glucose sensors based on ferrocene containing polymeric electron transfer systems-a preliminary report. AB - In this paper we introduce a preliminary work in which a redox mediator containing polyvinylferrocene immobilized on silica gel particles is synthesized. Carbon paste electrodes prepared with this polymeric mediator were studied for their stability in aqueous media for a month. The results indicated that approximately 70% of the initial activity was retained after a month. A steady state amperometric measurement of this carbon paste electrode containing this mediator and glucose oxidase at different potentials showed a linear relationship at elevated glucose concentration. PMID- 12782472 TI - Monitoring freshwater sediments. AB - The objectives of the SENSPOL Expert Meeting on 'Monitoring Freshwater Sediments' held in Antwerp, Belgium, 12-13 September, 2001, were firstly to identify and define problems and secondly to develop a realistic strategy to solve these problems. Both of the stakeholder groups (governmental authorities and the dredging industry) present at the workshop participated in detailed discussions to elucidate the role of sensors in the field of sediments and sediment/water interfaces. The 19 invited experts were agreed that in situ monitoring systems are needed to monitor freshwater sediments. New recognised tools for sediment monitoring would help industry to meet the governmental sediment quality criteria and to handle the data concerning historic river contamination and geological background data. The need to monitor by effect-related studies together with chemical monitoring was stressed. The main focus for development of new sensor tools should be for on site determination of certain priority pollutants where there would be advantage over existing methods or where no suitable method exists, and to monitor biological effects (alarm systems and effect-related on site tests). Sensing technologies would also be useful to monitor bioavailability in sediments in situ to provide information for risk assessment. In addition, they could be of use to monitor bioremediation in situ. A useful role was forseen in dredging sediments, for in situ sediment screening and to guide treatment of dredged material. The new sensing tools presented, included determination of metal concentrations in sediments using the diffuse gradients in thin films (DGT) technique (Lancaster University, UK), an analytical protocol for determination of metal speciation in sediments (Universitat Autonoma de Barcelona, Spain), microbiotests for determination of sediment toxicity (University of Ghent, Belgium), a portable whole cell sensors device for heavy metal bioavailability (VITO, Belgium) and a microfabricated sensor array system for Pb concentration profile measurement in the microM range at the liquid-solid interface (University of Geneva, Switzerland). PMID- 12782473 TI - MEAD (part II)-Predictions of radioactivity concentrations in the Irish Sea. AB - The predictions from MEAD, a model that simulates the transport of radionuclides in the marine environment, are presented for the Irish Sea. MEAD predictions for (137)Cs and Pu(alpha) are presented following discharges from BNFL Sellafield and the predictions compared to measured data from near the discharge location and further a field in the Irish Sea. The model performs well in most circumstances given the uncertainties involved in both modelling and data collection although some inconsistencies in the predictions are found. MEAD is also compared to other models of radionuclide transport in the Irish Sea. PMID- 12782474 TI - Formation of radioactivity enriched soils in mountain areas. AB - A field study was carried out in the Mercantour Mountains at 2200 m altitude to investigate the processes of soil enrichment in atmospheric Chernobyl (137)Cs. Soils with high (137)Cs activities have been collected in the pasture areas with frequently measured (137)Cs activity values of the order of 7000 Bq m(-2). At some single spots (about 6% of the studied area), activity in soils reached 300000 Bq m(-2), which represents 44% of the (137)Cs of the total area. Data further showed that spatial distribution of Cs depends widely on its origin: Chernobyl Cs is mainly concentrated in "enriched" soils, whereas older Cs and (241)Am fallout from nuclear weapons tests (NWTs) and natural atmospheric (210)Pb in soils is less heterogeneously distributed. In order to elucidate the processes which have led to the enrichment in Chernobyl (137)Cs in the Alps in May of 1986, we have studied the repartition of atmospheric (7)Be isotope (half-life=53.3 d) in the pasture compartments (soil, litter, grass, and snow). Snow (7)Be data give evidence that fallout enrichment is related to snow accumulation (snow drift). The transfer of beryllium occurs rapidly to the grass and litter, where the strongest pollutant accumulations were measured. However, (7)Be transport to the soil required more than 8 months. PMID- 12782475 TI - Genotoxicity and toxicity assay of water sampled from a radium production industry storage cell territory by means of Allium-test. AB - Water from natural reservoirs located near the radium production industry storage cell were analyzed using the anaphase-telophase chromosome aberration assay that was carried out on Allium schoenoprasum L. meristematic root tip cells. (262)Ra, (228)U, (232)Th, (210)Pb and (210)Po concentrations in all samples were found not to exceed the radioactivity concentration guides. The concentrations of 10 heavy metal ions were measured in water samples, but only Zn and Mn levels exceeded the maximum permissible concentration for the natural reservoirs. All water samples caused a significant increase of the chromosome aberration frequency as compared to control. The chromosome aberration spectrum analysis shows that the genotoxic effect was a result of chemical toxicity mainly. Two samples from the brook springhead were found to be toxic. The regression analysis results show that the mitotic index increased in parallel to Zn ion levels, and decreased with higher (238)U concentrations. The water samples genotoxicity positively correlated with the Zn concentration. The present work demonstrates that in order to achieve pollutant screening, it is not sufficient to determine the pollutants concentration only. Adequate conclusions on the risk due to environment contamination need to be based on the additional simultaneous use of toxicity and genotoxicity tests. When bioassays indicate some genotoxic and toxic effects, the determination of the chemical composition of the samples is then required. A combination of these two methods allows the identification of the elements that require constant biological monitoring. In the study reported here, those elements are Zn and (238)U. PMID- 12782476 TI - Anthropogenic radionuclides in the Japan Sea: their distributions and transport processes. AB - The anthropogenic radionuclides, (90)Sr, (137)Cs and (239+240)Pu, were measured in the water column of the Japan Sea/East Sea during 1997-2000. The vertical profiles of radionuclide concentrations showed: exponential decrease with depth for (90)Sr and (137)Cs, and surface minimum/subsurface maximum for (239+240)Pu. These results do not differ substantially from results reported previously. The area-averaged concentrations of radionuclides in the Japan Sea are higher than those found in the Northwest Pacific Ocean below surface layer showing the accumulation of the radionuclides in the deep waters in the Japan Sea. Concerning spatial distributions, the area of high (137)Cs inventory extends from the Japan Basin into the Yamato Basin. It is suggested that wintertime convection of water, occurring mainly in the Japan Basin, causes the radionuclides to sink. The nuclides then advect into the Yamato Basin after detouring around the Yamato Rise. PMID- 12782477 TI - Radon levels in Cyprus. AB - Radon levels in atmospheric and aquatic systems in Cyprus have recently been measured using the radon monitor Alpha Guard. Indoor and outdoor radon levels were obtained in situ, whereas analysis of radon concentrations in water was performed using tap and ground water samples collected from several areas of the island. The average value for outdoor and indoor radon concentration is 11+/-10 and 7+/-6 Bq m(-3), respectively, and for tap and ground water 0.4 Bq l(-1) and 1.4 Bq l(-1), respectively. From these data the annual dose equivalent of airborne radon to the Cypriot population is about 0.19 mSv y(-1), which is quite low compared to the total dose equivalent of natural and man-made ionising radiation in Cyprus. Radon levels in aquatic systems are relatively low due to an exhaustive utilisation of ground water resources and also to the increased input of desalinated sea water in the water distribution network and eventually into the ground water reservoirs. PMID- 12782478 TI - S-carboxymethylcysteine inhibits the attachment of Streptococcus pneumoniae to human pharyngeal epithelial cells. AB - Streptococcus pneumoniae causes respiratory and other invasive infections. Increased resistance of this bacterium to antibiotics necessitates new approaches to the treatment of infections. Attachment of bacteria to human pharyngeal epithelial cells is the initial step in the pathogenesis of infection and S carboxymethylcysteine (S-CMC) can modulate the attachment of Moraxella catarrhalis and nontypable Haemophilus influenzae to epithelial cells. Unlike these two, S. pneumoniae is gram-positive and has a well-defined capsule. Here we examined the effects of S-CMC on the attachment and detachment of S. pneumoniae to human pharyngeal epithelial cells in vitro. Treatment of these cells with S CMC significantly reduced the number of attached S. pneumoniae. S-CMC also resulted in a significant increase in the detachment of already attached S. pneumoniae to epithelial cells. In addition, treatment of S. pneumoniae with S CMC significantly reduced their ability to attach to epithelial cells, but not the number of viable bacteria. Our study shows that S-CMC modulates the attachment of S. pneumoniae to human pharyngeal epithelial cells by acting both on cells and bacteria. PMID- 12782479 TI - Prior exposure to Mannheimia haemolytica leukotoxin or LPS enhances beta(2) integrin expression by bovine neutrophils and augments LKT cytotoxicity. AB - Mannheimia (Pasteurella) haemolytica serotype1 produces a variety of virulence factors that play an important role during the pathogenesis of bovine pneumonic pasteurellosis. Among these, a leukotoxin (LKT) and lipopolysaccharide (LPS) are thought to be the primary virulence factors that contribute to the characteristic pathology of pasteurellosis. Recent evidence suggests that M. haemolytica LKT binding to bovine leukocytes is mediated by the beta(2)-integrin CD11a/CD18 (LFA 1), which subsequently induces activation and death of these cells. Exposure of bovine peripheral blood neutrophils (PMNs) to LKT or LPS induces expression of inflammatory cytokines, which in turn can increase LFA-1 expression and conformational activation. In this study we demonstrated, by flow cytometry and Western blot, that bovine PMNs increased their LFA-1 expression following in vitro exposure to M. haemolytica LKT and LPS. Increased LFA-1 expression by PMNs exposed to LKT and LPS was associated with increased LKT binding and cell death. The results of this study suggest that M. haemolytica LKT and LPS might cooperatively increase LFA-1 expression, and by so doing amplify the lung inflammation that characterizes bovine pasteurellosis. PMID- 12782480 TI - Pathogenesis of Acanthamoeba infections. AB - Acanthamoeba are free-living, harmless organisms, however, given the opportunity and the appropriate conditions, they can cause painful, sight-threatening as well as fatal infections and, thus, are considered opportunistic pathogens. Acanthamoeba infections have become increasingly important in the past few years due to increasing populations of contact lens users and AIDS patients. The mechanisms associated with the pathogenesis of Acanthamoeba tend to be highly complex, depending on parasite, host and the environmental factors. Elucidation of the biochemical, cellular and molecular basis of the pathogenesis of diseases caused by Acanthamoeba may lead to the development of therapeutic interventions. PMID- 12782481 TI - Functional characterization of HgbB, a new hemoglobin binding protein of Pasteurella multocida. AB - The biological function and role in pathogenesis of a Pasteurella multocida A:1 strain hemoglobin binding protein was investigated. The hgbB gene from the P. multocida A:1 strain, VP161, was cloned and characterized. hgbB was 2991 bp in length and encoded a mature length protein of 111 kDa. HgbB was predicted to be an outer membrane protein and shared 68 and 69% similarity to the hemoglobin/hemoglobin-haptoglobin binding protein, HI0712 from Haemophilus influenzae Rd and HgpC, from H. influenzae b, respectively. HgbB exhibited features typical of TonB dependent receptors, including seven conserved regions typical of these proteins, and conserved invariant residues. Escherichia coli expressing recombinant HgbB was found to bind hemoglobin in a solid phase dot blot binding assay. However, when a truncated form of the protein was expressed in E. coli, cells could no longer bind hemoglobin. Insertional inactivation of hgbB did not affect the ability of P. multocida to bind hemoglobin, nor its ability to produce disease in a mouse model. In addition, recombinant HgbB did not confer any protection against homologous or heterologous challenge. PMID- 12782482 TI - In vitro GAP activity towards RhoA, Rac1 and Cdc42 is not a prerequisite for YopE induced HeLa cell cytotoxicity. AB - The YopE cytotoxin of Yersinia is an essential virulence determinant that is translocated into the eukaryotic target cell via a plasmid-encoded type III secretion system. YopE possess a GTPase activating protein activity that in vitro has been shown to down regulate RhoA, Rac1, and Cdc42. Translocated YopE induces de-polymerisation of the actin microfilament structure in the eukaryotic cell which results in a rounding up of infected cells described as a cytotoxic effect. Here, we have investigated the importance of different regions of YopE for induction of cytotoxicity and in vitro GAP activity. Sequential removal of the N- and C-terminus of YopE identified the region between amino acids 90 and 215 to be necessary for induction of cytotoxicity. Internal deletions containing the essential arginine at position 144 resulted in a total loss of cytotoxic response. In-frame deletions flanking the arginine finger defined a region important for the cytotoxic effect to amino acids 166-183. Four triple-alanine substitution mutants in this region, YopE166-8A, 169-71A, 175-7A and 178-80A were still able to induce cytotoxicity on HeLa cells although they did not show any in vitro GAP activity towards RhoA, Rac1 or Cdc42. A substitution mutant in position 206-8A showed the same phenotype, ability to induce cytotoxic response but no in vitro GAP activity. We speculate that YopE may have additional unidentified targets within the eukaryotic cell. PMID- 12782485 TI - Interrelationships between the precautionary principle, prediction strategies, and sustainable use of the planet. AB - In this article, I examine the relationships between new concepts of human activity in the environment and several prevention strategies used to plot a course toward sustainable use. Natural capitalism and industrial ecology are relatively new concepts that provide a framework for environmental management. Although the precautionary principle puts into policy a determination to prevent environmental damage before it occurs, natural capitalism and industrial ecology go beyond the prevention of environmental damage to the optimization of environmental interactions. The risk assessment tools necessary for preventive management continue to be essential. However, additional tools are needed to go beyond prevention to optimization. A holistic, scientific approach to the human place within the environment is needed, including both interdisciplinary and large-scale research. PMID- 12782486 TI - Nerve cell damage in mammalian brain after exposure to microwaves from GSM mobile phones. AB - The possible risks of radio-frequency electromagnetic fields for the human body is a growing concern for our society. We have previously shown that weak pulsed microwaves give rise to a significant leakage of albumin through the blood-brain barrier. In this study we investigated whether a pathologic leakage across the blood-brain barrier might be combined with damage to the neurons. Three groups each of eight rats were exposed for 2 hr to Global System for Mobile Communications (GSM) mobile phone electromagnetic fields of different strengths. We found highly significant (p< 0.002) evidence for neuronal damage in the cortex, hippocampus, and basal ganglia in the brains of exposed rats. PMID- 12782487 TI - Crystallographic analysis of a hydroxylated polychlorinated biphenyl (OH-PCB) bound to the catalytic estrogen binding site of human estrogen sulfotransferase. AB - Certain hydroxylated polychlorinated biphenyls (OH-PCBs) inhibit the human estrogen sulfotransferase (hEST) at subnanomolar concentrations, suggesting a possible pathway for PCB toxicity due to environmental exposure in humans. To address the structural basis of the inhibition, we have determined the crystal structure of hEST in the presence of the sulfuryl donor product 3 phosphoadenosine 5 -phosphate and the OH-PCB 4,4 -OH 3,5,3,5 -tetraCB. The OH-PCB binds in the estrogen binding site with the position of the first phenolic ring in an orientation similar to the phenolic ring of 17 beta-estradiol. Interestingly, the OH-PCB does not bind in a planar conformation, but rather with a 30-degree twist between the phenyl rings. The crystal structure of hEST with the OH-PCB bound gives physical evidence that certain OH-PCBs can mimic binding of estrogenic compounds in biological systems. PMID- 12782488 TI - Association between arsenic exposure from a coal-burning power plant and urinary arsenic concentrations in Prievidza District, Slovakia. AB - To assess the arsenic exposure of a population living in the vicinity of a coal burning power plant with high arsenic emission in the Prievidza District, Slovakia, 548 spot urine samples were speciated for inorganic As (Asinorg), monomethylarsonic acid (MMA), dimethylarsinic acid (DMA), and their sum (Assum). The urine samples were collected from the population of a case-control study on nonmelanoma skin cancer (NMSC). A total of 411 samples with complete As speciations and sufficient urine quality and without fish consumption were used for statistical analysis. Although current environmental As exposure and urinary As concentrations were low (median As in soil within 5 km distance to the power plant, 41 micro g/g; median urinary Assum, 5.8 microg/L), there was a significant but weak association between As in soil and urinary Assum(r = 0.21, p < 0.01). We performed a multivariate regression analysis to calculate adjusted regression coefficients for environmental As exposure and other determinants of urinary As. Persons living in the vicinity of the plant had 27% higher Assum values (p < 0.01), based on elevated concentrations of the methylated species. A 32% increase of MMA occurred among subjects who consumed homegrown food (p < 0.001). NMSC cases had significantly higher levels of Assum, DMA, and Asinorg. The methylation index Asinorg/(MMA + DMA) was about 20% lower among cases (p < 0.05) and in men (p < 0.05) compared with controls and females, respectively. PMID- 12782489 TI - Influence of airspace geometry and surfactant on the retention of man-made vitreous fibers (MMVF 10a). AB - Inhaled and deposited man-made vitreous fibers (MMVF) 10a (low-fluorine preparation of Schuller 901 insulation glass) were studied by electron microscopy in hamster lungs, fixed by intravascular perfusion within 23 +/- 2 min (SD) of the initial inhalation. We found fibers on the surfaces of conducting airways and alveoli. In the airways, 89% of the fibers were totally and 11% partially covered by lining-layer material. In the alveoli, 32% of the fibers were totally submersed; others touched the alveolar wall, stuck at one end, bridging the airspace. Studies in a surface balance showed that fibers were immersed into the aqueous subphase by approximately 50% at film surface tensions of 20-25 mJ/m2) and were submersed (totally immersed; i.e., totally surrounded by fluid) at approximately 10 mJ/m2). Fibers were also found to be phagocytosed by macrophages. We found a substantial number of particle profiles within alveolar blood capillaries. Fiber length and alveolar geometry appear to be important limiting factors for the submersion of vitreous fibers into the lungs' surface lining layer. PMID- 12782490 TI - Airborne particles of the california central valley alter the lungs of healthy adult rats. AB - Epidemiologic studies have shown that airborne particulate matter (PM) with a mass median aerodynamic diameter < 10 microm (PM10) is associated with an increase in respiratory-related disease. However, there is a growing consensus that particles < 2.5 microm (PM2.5), including many in the ultrafine (< 0.1 microm) size range, may elicit greater adverse effects. PM is a complex mixture of organic and inorganic compounds; however, those components or properties responsible for biologic effects on the respiratory system have yet to be determined. During the fall and winter of 2000-2001, healthy adult Sprague-Dawley rats were exposed in six separate experiments to filtered air or combined fine (PM2.5) and ultrafine portions of ambient PM in Fresno, California, enhanced approximately 20-fold above outdoor levels. The intent of these studies was to determine if concentrated fine/ultrafine fractions of PM are cytotoxic and/or proinflammatory in the lungs of healthy adult rats. Exposures were for 4 hr/day for 3 consecutive days. The mean mass concentration of particles ranged from 190 to 847 microg/m3. PM was enriched primarily with ammonium nitrate, organic and elemental carbon, and metals. Viability of cells recovered by bronchoalveolar lavage (BAL) from rats exposed to concentrated PM was significantly decreased during 4 of 6 weeks, compared with rats exposed to filtered air (p< 0.05). Total numbers of BAL cells were increased during 1 week, and neutrophil numbers were increased during 2 weeks. These observations strongly suggest exposure to enhanced concentrations of ambient fine/ultrafine particles in Fresno is associated with mild, but significant, cellular effects in the lungs of healthy adult rats. PMID- 12782491 TI - Exposure assessment of particulate matter for susceptible populations in Seattle. AB - In this article we present results from a 2-year comprehensive exposure assessment study that examined the particulate matter (PM) exposures and health effects in 108 individuals with and without chronic obstructive pulmonary disease (COPD), coronary heart disease (CHD), and asthma. The average personal exposures to PM with aerodynamic diameters < 2.5 microm (PM2.5) were similar to the average outdoor PM2.5 concentrations but significantly higher than the average indoor concentrations. Personal PM2.5 exposures in our study groups were lower than those reported in other panel studies of susceptible populations. Indoor and outdoor PM2.5, PM10 (PM with aerodynamic diameters < 10 microm), and the ratio of PM2.5 to PM10 were significantly higher during the heating season. The increase in outdoor PM10 in winter was primarily due to an increase in the PM2.5 fraction. A similar seasonal variation was found for personal PM2.5. The high-risk subjects in our study engaged in an equal amount of dust-generating activities compared with the healthy elderly subjects. The children in the study experienced the highest indoor PM2.5 and PM10 concentrations. Personal PM2.5 exposures varied by study group, with elderly healthy and CHD subjects having the lowest exposures and asthmatic children having the highest exposures. Within study groups, the PM2.5 exposure varied depending on residence because of different particle infiltration efficiencies. Although we found a wide range of longitudinal correlations between central-site and personal PM2.5 measurements, the longitudinal r is closely related to the particle infiltration efficiency. PM2.5 exposures among the COPD and CHD subjects can be predicted with relatively good power with a microenvironmental model composed of three microenvironments. The prediction power is the lowest for the asthmatic children. PMID- 12782492 TI - Insecticidal juvenile hormone analogs stimulate the production of male offspring in the crustacean Daphnia magna. AB - Juvenile hormone analogs (JHAs) represent a class of insecticides that were designed specifically to disrupt endocrine-regulated processes relatively unique to insects. Recently we demonstrated that the crustacean juvenoid hormone methyl farnesoate programs oocytes of the crustacean Daphnia magna to develop into males. We hypothesized that insecticidal JHAs might mimic the action of methyl farnesoate, producing altered sex ratios of offspring. Daphnids were exposed chronically (3 weeks) to sublethal concentrations of methyl farnesoate, the JHA pyriproxyfen, and several nonjuvenoid chemicals to discern whether excess male offspring production is a generic response to stress or a specific response to juvenoid hormones. Only methyl farnesoate and pyriproxyfen increased the percentage of males produced by exposed maternal organisms. As previously reported with methyl farnesoate, acute exposure (24 hr) to either pyriproxyfen or the JHA methoprene caused oocytes maturing in the ovary to develop into males. We performed experiments to determine whether combined effects of a JHA and methyl farnesoate conformed better to a model of concentration addition (indicative of same mechanism of action) or independent joint action (indicative of different mechanisms of action). Combined effects conformed better to the concentration addition model, although some synergy, of unknown etiology, was evident between the insecticides and the hormone. These experiments demonstrate that insecticidal JHAs mimic the action of the crustacean juvenoid hormone methyl farnesoate, resulting in the inappropriate production of male offspring. The occurrence of such an effect in the environment could have dire consequences on susceptible crustacean populations. PMID- 12782493 TI - Cytopathology of the nasal mucosa in chronic exposure to diesel engine emission: a five-year survey of Swiss customs officers. AB - The simple and cheap technique of nasal cytology was used to assess possible adverse effects of chronic exposure to diesel engine emission (DEE) on respiratory mucous membranes. Brush cytology probes were taken from the noses of 194 male, nonsmoking customs officers twice a year (January and July) over a period of 5 years. The study group of 136 officers was solely occupied with clearing of diesel trucks (8.4 hr/day, 42 hr/week). Measured DEE concentrations varied between 31 and 60 microg/m3) and of benzo[a]pyrene concentrations were between 10 and 15 ng/m3). The control group of 58 officers worked only in the office. Over the 5-year period, similar results were obtained in summer and winter. In contrast to those not exposed to DEE, those who were had clear goblet cell hyperplasia with increased metaplastic and dysplastic epithelia and an increase in leukocytes. We found no evidence of progression of the cytopathologic changes. The findings may be described as a chronic inflammation of the nasal mucous membrane in the presence of chronic DEE exposure (chemical-induced rhinitis). Additionally, the findings of metaplastic and dysplastic nasal epithelia in the exposed subjects may indicate a genotoxic effect of chronic DEE exposure in humans. PMID- 12782494 TI - Increased concentrations of polychlorinated biphenyls, hexachlorobenzene, and chlordanes in mothers of men with testicular cancer. AB - An increasing incidence of testicular cancer has been reported from several countries in the Western world during the last decades. According to current hypothesis, testicular cancer is initiated during the fetal period, and exposure to endocrine disruptors, i.e., xenoestrogens, has been of concern. In this investigation we studied the concentrations of the sum of 38 polychlorinated biphenyls (PCBs), p,p'-dichlorodiphenyl-dichloroethylene, hexachlorobenzene (HCB), and chlordanes, in 61 cases with testicular cancer and 58 age-matched controls. Furthermore, case and control mothers were also asked to participate, and 44 case mothers and 45 control mothers agreed. They were of similar age. In cases only the concentration on lipid basis of cis-nonachlordane was significantly increased, whereas case mothers showed significantly increased concentrations of the sum of PCBs, HCB, trans- and cis-nonachlordane, and the sum of chlordanes. Among case mothers the sum of PCBs yielded an odds ratio (OR) of 3.8; 95% confidence interval (CI), 1.4-10 was calculated using the median concentration for the control mothers as cutoff value. For HCB, OR = 4.4 (95% CI, 1.7-12); for trans-nonachlordane, OR = 4.1 (95% CI, 1.5-11); for cis nonachlordane, OR = 3.1 (95% CI, 1.2-7.8); and for sum of chlordanes, OR = 1.9 (95% CI, 0.7-5.0). No consistent different risk pattern was found for seminoma or nonseminoma testicular cancer. PMID- 12782495 TI - Chlorination by-products in drinking water and menstrual cycle function. AB - We analyzed data from a prospective study of menstrual cycle function and early pregnancy loss to explore further the effects of trihalomethanes (THM) on reproductive end points. Premenopausal women ((italic)n(/italic) = 403) collected urine samples daily during an average of 5.6 cycles for measurement of steroid metabolites that were used to define menstrual parameters such as cycle and phase length. Women were asked about consumption of various types of water as well as other habits and demographics. A THM level was estimated for each cycle based on residence and quarterly measurements made by water utilities during a 90-day period beginning 60 days before the cycle start date. We found a monotonic decrease in mean cycle length with increasing total THM (TTHM) level; at > 60 microg/L, the adjusted decrement was 1.1 days [95% confidence interval (CI), -1.8 to -0.40], compared with less than or equal to 40 microg/L. This finding was also reflected as a reduced follicular phase length (difference -0.94 day; 95% CI, 1.6 to -0.24). A decrement in cycle and follicular phase length of 0.18 days (95% CI, -0.29 to -0.07) per 10 microg/L unit increase in TTHM concentration was found. There was little association with luteal phase length, menses length, or cycle variability. Examining the individual THMs by quartile, we found the greatest association with chlorodibromomethane or the sum of the brominated compounds. Incorporating tap water consumption showed a similar pattern of reduced cycle length with increasing TTHM exposure. These findings suggest that THM exposure may affect ovarian function and should be confirmed in other studies. PMID- 12782496 TI - Disparities in exposure to air pollution during pregnancy. AB - Previous research shows poorer birth outcomes for racial and ethnic minorities and for persons with low socioeconomic status (SES). We evaluated whether mothers in groups at higher risk for poor birth outcomes live in areas of higher air pollution and whether higher exposure to air pollution contributes to poor birth outcomes. An index representing long-term exposure to criteria air pollutants was matched with birth certificate data at the county level for the United States in 1998-1999. We used linear regression to estimate associations between the air pollution index and maternal race and educational attainment, a marker for SES of the mother, controlling for age, parity, marital status, and region of the country. Then we used logistic regression models both to estimate likelihood of living in counties with the highest levels of air pollution for different racial groups and by educational attainment, adjusting for other maternal risk factors, and to estimate the effect of living in counties with higher levels of air pollution on preterm delivery and births small for gestational age (SGA). Hispanic, African-American, and Asian/Pacific Islander mothers experienced higher mean levels of air pollution and were more than twice as likely to live in the most polluted counties compared with white mothers after controlling for maternal risk factors, region, and educational status [Hispanic mothers: adjusted odds ratio (AOR) = 4.66; 95% confidence interval (95% CI), 1.92-11.32; African American mothers: AOR = 2.58; 95% CI, 1.00-6.62; Asian/Pacific Islander mothers: AOR = 2.82; 95% CI, 1.07-7.39]. Educational attainment was not associated with living in counties with highest levels of the air pollution index (AOR = 0.95; 95% CI, 0.40-2.26) after adjusting for maternal risk factors, region of the country, and race/ethnicity. There was a small increase in the odds of preterm delivery (AOR = 1.05; 95% CI, 0.99-1.12) but not SGA (AOR = 0.96; 95% CI, 0.86 1.07) in a county with high air pollution. Additional risk of residing in areas with poor air quality may exacerbate health problems of infants and children already at increased risk for poor health. PMID- 12782497 TI - Maternal serum dioxin levels and birth outcomes in women of Seveso, Italy. AB - 2,3,7,8-Tetrachlorodibenzo-(italic)para(/italic)-dioxin (TCDD), a ubiquitous environmental contaminant, is associated with increased fetal loss and reduced birth weight in animal studies. In 1976, an explosion at a trichlorophenol plant near Seveso, Italy, resulted in the highest TCDD exposure known in human residential populations. In 1996, we initiated the Seveso Women's Health Study, a retrospective cohort study of women who resided in the most contaminated areas, zones A and B. We examined the relation of pregnancy outcome in 510 women (888 total pregnancies) to maternal TCDD levels measured in serum collected shortly after the explosion. Ninety-seven pregnancies (10.9%) ended as spontaneous abortions (SABs). There was no association of log(subscript)10(/subscript) TCDD with SAB [adjusted odds ratio (OR) = 0.8; 95% confidence interval (CI), 0.6-1.2], with birth weight (adjusted beta = -4 g; 95% CI, -68 to 60), or with births that were small for gestational age (SGA) (adjusted OR = 1.2; 95% CI, 0.8-1.8). However, associations with birth weight (adjusted beta = -92 g; 95% CI, -204 to 19) and with SGA (adjusted OR = 1.4; 95% CI, 0.6-2.9) were stronger for pregnancies within the first 8 years after exposure. TCDD was associated with a 1.0-1.3 day nonsignificant adjusted decrease in gestational age and a 20-50% nonsignificant increase in the odds of preterm delivery. It remains possible that the effects of TCDD on birth outcomes are yet to be observed, because the most heavily exposed women in Seveso were the youngest and the least likely to have yet had a pregnancy. PMID- 12782499 TI - Childhood leukemia: electric and magnetic fields as possible risk factors. AB - Numerous epidemiologic studies have reported associations between measures of power-line electric or magnetic fields (EMFs) and childhood leukemia. The basis for such associations remains unexplained. In children, acute lymphoblastic leukemia represents approximately three-quarters of all U.S. leukemia types. Some risk factors for childhood leukemia have been established, and others are suspected. Pathogenesis, as investigated in animal models, is consistent with the multistep model of acute leukemia development. Studies of carcinogenicity in animals, however, are overwhelmingly negative and do not support the hypothesis that EMF exposure is a significant risk factor for hematopoietic neoplasia. We may fail to observe effects from EMFs because, from a mechanistic perspective, the effects of EMFs on biology are very weak. Cells and organs function despite many sources of chemical "noise" (e.g., stochastic, temperature, concentration, mechanical, and electrical noise), which exceed the induced EMF "signal" by a large factor. However, the inability to detect EMF effects in bioassay systems may be caused by the choice made for "EMF exposure." "Contact currents" or "contact voltages" have been proposed as a novel exposure metric, because their magnitude is related to measured power-line magnetic fields. A contact current occurs when a person touches two conductive surfaces at different voltages. Modeled analyses support contact currents as a plausible metric because of correlations with residential magnetic fields and opportunity for exposure. The possible role of contact currents as an explanatory variable in the reported associations between EMFs and childhood leukemia will need to be clarified by further measurements, biophysical analyses, bioassay studies, and epidemiology. PMID- 12782500 TI - World Trade Center fine particulate matter--chemistry and toxic respiratory effects: an overview. AB - The 11 September 2001 terrorist attack on New York City's World Trade Center (WTC) caused an unprecedented environmental emergency. The collapse of the towers sent a tremendous cloud of crushed building materials and other pollutants into the air of lower Manhattan. In response to the calamity, federal, state, and city environmental authorities and research institutes devoted enormous resources to evaluate the impact of WTC-derived air pollution on public health. Unfortunately, on the day of the disaster, no air-sampling monitors were operating close to the WTC site to characterize and quantify pollutants in the dust cloud. However, analysis of fallen dust samples collected 5 and 6 days after the attack showed that 1-4% by weight consisted of particles small enough to be respirable (Lioy et al. 2002). These particles included fine particulate matter, or PM(subscript)2.5(/subscript) [PM < 2.5 micro m mass median aerodynamic diameter (MMAD)], which can be inhaled deep into the lung and is associated with cardiovascular and respiratory health effects. Because of the extremely high concentrations of dust immediately after the collapse of the towers, even a relatively small proportion of PM(subscript)2.5(/subscript) in the dust clouds could have contributed to breathing problems in rescue workers and others who were not wearing protective masks. PMID- 12782498 TI - Organochlorines, lead, and mercury in Akwesasne Mohawk youth. AB - Most humans have detectable body burdens of polychlorinated biphenyls (PCBs), hexachlorobenzene (HCB), and p,p'-dichlorophenyldichloroethylene p,p'-DDE), a metabolite of p,p'-dichlorodiphenyltrichloroethane (DDT). Native American communities may be at increased risk of exposure through subsistence-based diets and greater physical contact with contaminated soil and water. In this article we describe the levels of toxicants (PCBs, p,p'-DDE, HCB, mirex, lead, and mercury) among youth 10-17 years old (n = 271) of the Akwesasne Mohawk Nation. Ultratrace, congener-specific PCB analysis of human serum quantitated 83 PCB congeners (plus 18 as pairs/triplets), in addition to p,p'-DDE, HCB, and mirex, and included all major Aroclor-derived congeners typically present in human samples. Twenty congeners (in 16 chromatographic peaks) were detected in 50% or more of the individuals sampled [geometric mean (GM) of the sum of these congeners = 0.66 ppb]. Thirteen congeners (in 10 peaks) were detected in 75% or more of the samples (GM = 0.51 ppb). Of the 20 congeners detected in 50% or more of the samples, 17 had five or more chlorine substitutions. International Union for Pure and Applied Chemistry congeners 118, 101(+90), and 153 were detected in nearly all participants (GM = 0.06 ppb, 0.05 ppb, 0.09 ppb, respectively). p,p'-DDE and HCB were detected in 100% and 98% of the samples (GM: p,p'-DDE = 0.37 ppb; HCB = 0.03 ppb). Mirex was detected in approximately 46% of the samples (GM = 0.02 ppb). No cases of elevated lead level were observed. One participant had a mercury level marginally higher than the U.S. Environmental Protection Agency's current level of concern (0.50 microg/dL). Although differences in analytic methods and participant ages limit comparability, toxicant levels from the Mohawk youth are lower than those associated with severe food contamination (Yusho and Yu-cheng) but similar to other chronically exposed groups. PMID- 12782501 TI - Chemical analysis of World Trade Center fine particulate matter for use in toxicologic assessment. AB - The catastrophic destruction of the World Trade Center (WTC) on 11 September 2001 caused the release of high levels of airborne pollutants into the local environment. To assess the toxicity of fine particulate matter [particulate matter with a mass median aerodynamic diameter < 2.5 microm (PM2.5)], which may adversely affect the health of workers and residents in the area, we collected fallen dust samples on 12 and 13 September 2001 from sites within a half-mile of Ground Zero. Samples of WTC dust were sieved, aerosolized, and size-separated, and the PM2.5 fraction was isolated on filters. Here we report the chemical and physical properties of PM2.5 derived from these samples and compare them with PM2.5 fractions of three reference materials that range in toxicity from relatively inert to acutely toxic (Mt. St. Helens PM; Washington, DC, ambient air PM; and residual oil fly ash). X-ray diffraction of very coarse sieved WTC PM (< 53 microm) identified calcium sulfate (gypsum) and calcium carbonate (calcite) as major components. Scanning electron microscopy confirmed that calcium-sulfur and calcium-carbon particles were also present in the WTC PM2.5 fraction. Analysis of WTC PM2.5 using X-ray fluorescence, neutron activation analysis, and inductively coupled plasma spectrometry showed high levels of calcium (range, 22-33%) and sulfur (37-43% as sulfate) and much lower levels of transition metals and other elements. Aqueous extracts of WTC PM2.5 were basic (pH range, 8.9-10.0) and had no evidence of significant bacterial contamination. Levels of carbon were relatively low, suggesting that combustion-derived particles did not form a significant fraction of these samples recovered in the immediate aftermath of the destruction of the towers. Because gypsum and calcite are known to cause irritation of the mucus membranes of the eyes and respiratory tract, inhalation of high doses of WTC PM2.5 could potentially cause toxic respiratory effects. PMID- 12782503 TI - Restoring the foundation: Tracking chemical exposures and human health. PMID- 12782502 TI - World Trade Center fine particulate matter causes respiratory tract hyperresponsiveness in mice. AB - Pollutants originating from the destruction of the World Trade Center (WTC) in New York City on 11 September 2001 have been reported to cause adverse respiratory responses in rescue workers and nearby residents. We examined whether WTC-derived fine particulate matter [particulate matter with a mass median aerodynamic diameter < 2.5 microm (PM2.5)] has detrimental respiratory effects in mice to contribute to the risk assessment of WTC-derived pollutants. Samples of WTC PM2.5 were derived from settled dust collected at several locations around Ground Zero on 12 and 13 September 2001. Aspirated samples of WTC PM2.5 induced mild to moderate degrees of pulmonary inflammation 1 day after exposure but only at a relatively high dose (100 microg). This response was not as great as that caused by 100 microg PM2.5 derived from residual oil fly ash (ROFA) or Washington, DC, ambient air PM [National Institute of Standards and Technology, Standard Reference Material (SRM) 1649a]. However, this same dose of WTC PM2.5 caused airway hyperresponsiveness to methacholine aerosol comparable to that from SRM 1649a and to a greater degree than that from ROFA. Mice exposed to lower doses by aspiration or inhalation exposure did not develop significant inflammation or hyperresponsiveness. These results show that exposure to high levels of WTC PM2.5 can promote mechanisms of airflow obstruction in mice. Airborne concentrations of WTC PM2.5 that would cause comparable doses in people are high (approximately 425 microg/m3 for 8 hr) but conceivable in the aftermath of the collapse of the towers when rescue and salvage efforts were in effect. We conclude that a high-level exposure to WTC PM2.5 could cause pulmonary inflammation and airway hyperresponsiveness in people. The effects of chronic exposures to lower levels of WTC PM2.5, the persistence of any respiratory effects, and the effects of coarser WTC PM are unknown and were not examined in these studies. Degree of exposure and respiratory protection, individual differences in sensitivity to WTC PM2.5, and species differences in responses must be considered in assessing the risks of exposure to WTC PM2.5. PMID- 12782504 TI - The environment as a cornerstone of public health. PMID- 12782505 TI - Critique of Vreugdenhil et al.'s study linking PCBs to the play behaviors of Dutch girls and boys. PMID- 12782507 TI - Dutch girls and boys, PCB levels, and play behavior: what do the data really tell us? PMID- 12782508 TI - Adverse health effects of bisphenol A in early life. PMID- 12782513 TI - Body of evidence. PMID- 12782514 TI - Building civic health. PMID- 12782515 TI - The risk where you live. PMID- 12782517 TI - Effect of a delta receptor agonist on duration of survival during hemorrhagic shock. AB - OBJECTIVES: Selective delta receptor agonists have been shown to stabilize membrane physiologic processes, reduce metabolic rates, and provide protection against ischemic insults through K(ATP) channel opening in a variety of organ beds. However, their potential for affecting outcomes in states of generalized ischemia has not been explored. The authors examined the effect of the nonselective delta receptor agonist, DADLE (D-Ala2-Leu5-enkephalin), on hemodynamic stability and duration of survival in an animal model of severe hemorrhagic shock. METHODS: Conscious Sprague Dawley rats with indwelling catheters were hemorrhaged at a rate of 3.25 mL/100 grams over 20 minutes after half of the group received 1% DADLE (1 mg/kg IV). Following the hemorrhage, all rats were continuously monitored for heart rate (HR), mean arterial pressure (MAP), and life signs for up to three hours (death defined as apnea, systolic blood pressure < 30 mm Hg without pulsations, and electroencephalographic silence). Survival rates and hemodynamic trends were compared between the control and DADLE-treated groups. RESULTS: In the 14 rats studied (8 DADLE; 6 controls), initial hemorrhage resulted in similar hemodynamic shock (average MAP fall: 118 to 59 vs 119 to 55 mm Hg). Analysis of survival at 3.5 hours revealed statistically significant differences between the control and DADLE groups. While 50% of the DADLE group survived past the three hours, no control animals were still alive at the end of the experimental period. The MAP trended downward and the HR increased for the control group, but all hemodynamic parameters stabilized in the rats treated with DADLE. CONCLUSIONS: Most current strategies for treating shock focus on the supply side of resuscitation. The coordinated various actions of DADLE have the potential to work in concert in the intact organism to improve overall survival during severe hemorrhagic shock. In an animal model of severe hemorrhagic shock, there was improvement in hemodynamic stability and a prolonged survival with DADLE treatment. Physiologic manipulation with DADLE appears to be a way to improve survival during shock with possible clinical implications. PMID- 12782518 TI - Evaluation of a structured application assessment instrument for assessing applications to Canadian postgraduate training programs in emergency medicine. AB - OBJECTIVE: To determine the interrater reliability and predictive validity of a structured instrument for assessing applications submitted to a Fellow of the Royal College of Physicians of Canada (FRCP) emergency medicine residency program. METHODS: An application assessment instrument was derived based on faculty and resident input, institutional and national documents, and previous protocols. The instrument provided a score based on objective anchors for each of four application components. Three assessors were introduced to the instrument in a detailed tutorial session. Assessors were given five applications to score and results were compared for understanding of the scoring principles. The instrument was used in a developmental pilot to assess the 2001 cohort of applications and revised again. Applications for the 2002 study cohort were submitted through a central application service. Assessors used the instrument to score each application independently. Interrater reliability was determined by calculating a two-way mixed-effect Cronbach's alpha. RESULTS: Forty applications were received for the year 2002. Thirty-eight application packages were complete and data collection was complete for all 38. The single-rater reliabilities for the curriculum vitae, personal letter, transcript, reference letters, and overall package were 0.73, 0.52, 0.64, 0.61, and 0.72, respectively. The three-rater reliabilities for the components were 0.89, 0.77, 0.84, and 0.82, respectively. The three-rater reliability of the overall application score was 0.89. CONCLUSIONS: Three-rater reliabilities for each component and the entire application package were high. Multiple assessors are required to generate acceptable reliabilities. Using strict design and implementation principles can lead to a reliable instrument for assessing complex application packages. PMID- 12782519 TI - Emergency physicians' perspectives on smallpox vaccination. AB - OBJECTIVE: To evaluate emergency physician (EP) attitudes toward smallpox vaccination, the treatment of patients with suspected smallpox, and the threat of a bioterrorist attack. METHODS: This was a prospective study utilizing a standardized survey instrument that was distributed on November 16, 2002, and collected by February 1, 2003. EPs from a sample of 50 accredited emergency medicine programs were surveyed regarding their perspectives on smallpox vaccination. RESULTS: A total of 989 surveys were collected from 42 emergency medicine programs. Of the respondents, 43.4% would currently volunteer for smallpox vaccination. EPs previously vaccinated against smallpox were 1.46 times more likely to volunteer for vaccination (95% CI = 1.14 to 1.93). EPs who believed they were at risk for complications were less than half as likely to volunteer for vaccination. EPs who perceived a significant risk of a bioterrorist attack were 2.7 times more likely to volunteer for the vaccine compared with those who thought the risk was minimal (95% CI = 2.06 to 3.47). Of the respondents, 34.4% believed the risks of the vaccination outweighed the benefits, 33% did not, and 32.6% were unsure. CONCLUSIONS: Currently, fewer than half of EPs surveyed would volunteer for smallpox vaccination. Factors associated with a willingness to be vaccinated include previous smallpox vaccination and the perceived threat of a bioterrorist attack. The variation in EP attitudes toward smallpox vaccination may be due to uncertain risk-to-benefit ratio. The opinions and actions of EPs may be influential on current and future government policy and public opinion. PMID- 12782520 TI - Smallpox vaccination: a national survey of emergency health care providers. AB - Concerns about bioterrorism have prompted a national voluntary smallpox (SP) vaccination program in the United States. Although emergency health care providers are among the first targeted for vaccination, little is known about how these providers view the risks and benefits of SP vaccination. OBJECTIVES: To assess the willingness of emergency health care personnel to receive pre-event SP vaccination prior to the start of the national program. METHODS: The authors conducted a national cross-sectional, anonymous survey of 1,701 emergency physicians, nurses, and mid-level practitioners working full time in 13 adult and pediatric academic emergency departments in large U.S. cities in November and December 2002. The main outcome measure was willingness to be vaccinated against SP. Secondary outcomes included the prevalence of self-reported contraindications, and reasons for and against vaccination. RESULTS: 732 emergency health care providers returned questionnaires (response rate 43%). Overall, 73% (95% CI = 66% to 80%) were willing to receive pre-event SP vaccination. 18% (95% CI = 14% to 23%) reported contraindications to vaccination, and 50% (95% CI = 39% to 61%) of these providers were willing to receive pre event SP vaccination. Self-protection (72%) was the most common reason cited for desiring vaccination against SP; concern about vaccine-related adverse events (54%) was the most common reason cited for not wanting immunization. CONCLUSIONS: Most emergency health care providers express a willingness to receive pre-event SP immunization; self-protection is a principal motivating reason. A subset of health care providers, however, may place themselves at increased risk by desiring vaccination despite contraindications. PMID- 12782521 TI - Does the sedative agent facilitate emergency rapid sequence intubation? AB - OBJECTIVES: To ascertain whether the sedative agent administered during neuromuscular-blocking agent-facilitated intubation (rapid sequence intubation [RSI]) influences the number of attempts and overall success at RSI. METHODS: Records were drawn from an ongoing, prospective multicenter registry of emergency department intubations. Conditional logistic regression stratified by institution was used to identify factors associated with multiple intubation attempts and unsuccessful RSI. RESULTS: Of 3,407 intubations over 33 months in 22 institutions, 2,380 involved RSI. After correcting for the specialty and experience of the intubator and for the presence of airway aberrancy, the sedative agent was significantly associated with the number of attempts at intubation (p = 0.002). Specifically, the use of etomidate (adjusted odds ratio [OR] 0.35 [95% CI = 0.17 to 0.72]), ketamine (OR 0.27 [95% CI = 0.11 to 0.65]), a benzodiazepine (OR 0.47 [95% CI = 0.23 to 0.95]), or no sedative agent (OR 0.51 [95% CI = 0.23 to 1.13]) prior to neuromuscular blockade was associated with a lower likelihood of successful intubation on the first attempt, as compared with thiopental, methohexital, or propofol. The adjusted odds ratios for the likelihood of overall success had similar point estimates, but did not reach statistical significance due to lack of power (p = 0.2, with 36 unsuccessful intubations). Among patients receiving etomidate, intubation was more likely to be successful on the first attempt with increasing doses of either etomidate or succinylcholine. CONCLUSIONS: Thiopental, methohexital, and propofol appear to facilitate RSI in emergency department patients, independent of patient characteristics or intubator training. A deeper plane of anesthesia may improve intubating conditions in emergency patients undergoing RSI by complementing incomplete muscle paralysis. PMID- 12782522 TI - Is the ACLS score a valid prediction rule for survival after cardiac arrest? AB - The ACLS (advanced cardiac life support) Score was previously developed to predict survival from out-of-hospital cardiac arrest. Whether the arrest was witnessed, initial cardiac rhythm, performance of bystander cardiopulmonary resuscitation (CPR), and the response time of the paramedic unit were determined to be predictive of survival. However, the ACLS Score has not been validated in other emergency medical services systems. OBJECTIVES: The purpose of this study was to externally validate the ACLS Score in one patient population. METHODS: This was a retrospective cohort study performed at an urban county teaching hospital. The study population consisted of consecutive adult patients treated for out-of-hospital, nontraumatic cardiac arrest, and transported to the authors' institution between November 1, 1994, and September 30, 2001. Patient records for all cardiac arrests during the study period were reviewed. Study variables included witnessed arrest, initial arrest rhythm, bystander CPR, paramedic response time, and survival to hospital discharge. Predicted probability of survival to hospital discharge was calculated for each patient using the ACLS Score. The overall predicted and observed survival rates were compared using Flora's Z score. The Hosmer-Lemeshow test was used to evaluate the model's goodness-of-fit over a range of survival probabilities. RESULTS: Of 754 cardiac arrest patients enrolled in the study period, 575 (76%) patients had documentation that allowed scoring using the ACLS Score. Twenty-five (4%) patients survived to hospital discharge. The predicted number of survivors based on the ACLS Score was 104 (18%), yielding a Flora's Z statistic of -4.46 (p < 0.0001). After categorizing predicted survival probabilities into four categories, the resulting Hosmer-Lemeshow statistic was 210 (p << 10(-6)). Both goodness-of-fit statistics demonstrated extremely poor fit of the model. A receiver operating characteristic (ROC) curve was created, yielding an area under the ROC curve of 0.33 (95% CI = 0.19 to 0.47), signifying extremely poor discrimination. CONCLUSIONS: The previously published ACLS Score was not valid when applied to an external cohort of out-of-hospital cardiac arrest patients. An externally valid model is needed to predict survival to hospital discharge following out-of-hospital cardiac arrest. PMID- 12782523 TI - Acute pain is underassessed in out-of-hospital emergencies. AB - OBJECTIVES: To evaluate the quality of pain assessment by emergency medical services (EMS) in out-of-hospital emergencies. METHODS: A prospective study was conducted on a convenience sample of patients during a one-year observation period. Pain ratings assessed by emergency patients were documented at three different intervals during the emergency call, and compared with concomitant assessments by EMS providers. A visual analog scale (VAS) and a verbal pain scale (VPS) were used for pain assessment. Repeated-measures ANOVA and Dunnett's t-test were used for data analysis. RESULTS: Fifty-one out of 70 eligible patients met inclusion criteria. In most emergency patients the intensity of pain was underestimated by EMS, especially when pain was severe (p = 0.0001). During the course of transport, both pain and pain assessment by EMS improved significantly (p = 0.0001). The VAS and VPS were significantly correlated (p = 0.0001). CONCLUSIONS: EMS providers significantly underestimate their patients' pain severity. EMS providers should be more attentive to their patients' complaints and comfort. PMID- 12782524 TI - Evaluating paramedic comfort with field pronouncement: development and validation of an outcome measure. AB - OBJECTIVES: Interventions designed to improve cardiac resuscitation and the quality of field pronouncement need to consider outcomes on paramedic providers. The authors developed and evaluated the reliability and validity of a survey instrument measuring paramedic comfort with field pronouncement. METHODS: A mail survey of 120 paramedics (EMT-Ps) was performed using the Modified Dillman survey methodology. Questions were sorted for analysis into subgroups assessing psychological comfort and technical skills. Sixty-five respondents were retested within two weeks. RESULTS: The overall response rate was 96% (115). Respondents had an average age of 36 years (SD +/- 5), with 5.2 years (SD +/- 3.8) of out-of hospital experience as an EMT-P, and were involved in a median of ten field pronouncements annually (range = 2-60). The face and content validity of the survey instrument was consistent with a content matrix derived by a focus group. The Cronbach's alpha for the survey instrument was 0.91. The retest response rate was 76% (46). The test-retest reliability coefficient was 0.84. CONCLUSIONS: This survey is a valid and reliable instrument for measuring the paramedic psychological comfort with field pronouncement. The high response rate and intrareliability support its generalizability. This outcome measure may be helpful in evaluating the psychological impact of changes to emergency medical services (EMS) policy with respect to termination of resuscitation promoted by the National Association of EMS Physicians. PMID- 12782526 TI - The challenge of defining the "science" of airway management--what is the right outcome measure? PMID- 12782525 TI - Bispectral electroencephalographic analysis of patients undergoing procedural sedation in the emergency department. AB - OBJECTIVE: To determine whether there is a correlation between the level of sedation achieved during procedural sedation (PS) in the emergency department as determined by bispectral electroencephalographic (EEG) analysis (BIS) and the rate of respiratory depression (RD), the patient's perception of pain, recall of the procedure, and satisfaction. METHODS: This was a prospective observational study conducted in an urban county hospital of adult patients undergoing PS using propofol, methohexital, etomidate, and the combination of fentanyl and midazolam. Consenting patients were monitored by vital signs, pulse oximetry, nasal-sample end-tidal carbon dioxide (ETCO(2)), and BIS monitors during PS. Respiratory depression (RD) was defined as an oxygen saturation <90%, a change from baseline ETCO(2) of >10 mm Hg, or an absent ETCO(2) waveform at any time during the procedure. After the procedure, patients were asked to complete three 100-mm visual analog scales (VASs) concerning their perception of pain, recall of the procedure, and satisfaction with the procedure. Patients were divided into four groups based on the lowest BIS score recorded during the procedure, group 1, >85; group 2, 70-85; group 3, 60-69; group 4, <60. Rates of RD and VAS outcomes were compared between groups using chi-square statistics. RESULTS: One hundred eight patients were enrolled in the study. No serious adverse events were noted. RD was seen in three of 14 (21.4%) of the patients in group 1, seven of 34 (20.6%) in group 2, 16 of 26 (61.5%) in group 3, and 18 of 34 (52.9%) in group 4. The rate of RD in patients in group 2 was not significantly different from that in group 1 (p = 0.46). The rate of RD in group 2 was significantly lower than that in groups 3 (p = 0.0003) and 4 (p = 0.006). For the VAS data, when group 1 was compared with the combined groups 2, 3, and 4, it had significantly higher rates of pain (p = 0.003) and recall (p = 0.001), and a dissatisfaction rate (p = 0.085) that approached significance. When groups 2, 3, and 4 were compared with chi-square test, there was not a significant difference in pain (p = 0.151), recall (p = 0.27), or satisfaction (p = 0.25). CONCLUSIONS: Patients with a lowest recorded BIS score between 70 and 85 had the same VAS outcomes as more deeply sedated patients and the same rate of RD as less deeply sedated patients. This range of scores represented the optimally sedated patients in this study. PMID- 12782527 TI - "All the king's horses and all the king's men. . .". PMID- 12782528 TI - Site management of health issues in the 2001 World Trade Center disaster. AB - The terrorist destruction of the World Trade Center led to the greatest loss of life from a criminal incident in the history of the United States. There were 2,801 persons killed or missing at the disaster site, including 147 dead on two hijacked aircraft. Hundreds of buildings sustained direct damage or contamination. Forty different agencies responded with command and control exercised by an incident command system as well as an emergency operations center. Dozens of hazards complicated relief and recovery efforts. Five victims were rescued from the rubble. Up to 1,000 personnel worked daily at the World Trade Center disaster site. These workers collectively made an average of 270 daily presentations to health care providers in the first month post-disaster. Of presentations for clinical symptoms, leading clinical diagnoses were ocular injuries, headaches, and lung injuries. Mechanical injury accounted for 39% of clinical presentations and appeared preventable by personal protective equipment. Limitations emerged in the site application of emergency triage and clinical care. Notable assets in the site management of health issues include action plans from the incident command system, geographic information system products, wireless application technology, technical consensus among health and safety authorities, and workers' respite care. PMID- 12782530 TI - The Pediatric Emergency Care Applied Research Network (PECARN): rationale, development, and first steps. AB - Since its formal recognition as a medical specialty, the field of pediatric emergency medicine has made substantial advances with respect to its scope and sophistication. These advances have occurred in clinical practice as well as in the research base to improve clinical practice. There remain, however, many areas in emergency medical services for children (EMSC), in the out-of-hospital as well as the emergency department (ED) and hospital settings, that suffer from a lack of data to guide practice. In an effort to expand the quality and quantity of research in pediatric emergency care, the Pediatric Emergency Care Applied Research Network (PECARN) was created in October 2001. PECARN is the first federally funded national network for research in EMSC. PECARN is the result of Cooperative Agreement grants funded through the Health Resources and Services Administration (HRSA) with the purpose of developing an infrastructure capable of overcoming inherent barriers to pediatric EMSC research. Among these recognized barriers are low incidence rates of serious pediatric emergency events, the need for large numbers of children from varied backgrounds to achieve broadly representative study samples, lack of an infrastructure to test the efficacy of pediatric emergency care, and the need for a mechanism to translate study results into clinical practice. PECARN will serve as a national platform for collaborative research involving the continuum of care within the EMSC system, including out-of-hospital care, patient transport, ED and in-hospital care, and rehabilitation. This article describes the history of EMSC, the need for a national collaborative research network in EMSC, the organization and development of PECARN, and the work plan for the Network. PMID- 12782531 TI - Bench to bedside: electrophysiologic and clinical principles of noninvasive hemodynamic monitoring using impedance cardiography. AB - The evaluation of the hemodynamic state of the severely ill patient is a common problem in emergency medicine. While conventional vital signs offer some insight into delineating the circulatory pathophysiology, it is often impossible to determine the true clinical state from an analysis of blood pressure and heart rate alone. Cardiac output measurements by thermodilution have been the criterion standard for the evaluation of hemodynamics. However, this technology is invasive, expensive, time-consuming, and impractical for most emergency department environments. Impedance cardiography (ICG) is a noninvasive method of obtaining continuous measurements of hemodynamic data such as cardiac output that requires little technical expertise. ICG technology was first developed by NASA in the 1960s and is based on the idea that the human thorax is electrically a nonhomogeneous, bulk conductor. Variation in the impedance to flow of a high frequency, low-magnitude alternating current across the thorax results in the generation of a measured waveform from which stroke volume can be calculated by a modification of the pulse contour method. To adequately judge the possible role of this technology in the practice of emergency medicine, it is important to have a sufficient understanding of the basic scientific principles involved as well as the clinical validity and limitations of the technique. PMID- 12782532 TI - Smallpox vaccination for emergency physicians. AB - On December 13, 2002, President Bush formally announced the national smallpox vaccination program. The plan involves vaccinating health care workers who would respond to possible smallpox cases, including emergency physicians. Although not all aspects of the program are clearly established, it appears that vaccination of health care workers will occur in the near future. This joint statement has been used by the American Academy of Emergency Medicine and the Society for Academic Emergency Medicine regarding smallpox vaccination for emergency physicians. PMID- 12782533 TI - Emergency medicine animal research: does use of randomization and blinding affect the results? AB - OBJECTIVES: It has been shown that human clinical trials that lack randomization (RND) or blinding (BLD) often overestimate the magnitude of treatment effects. However, no studies have evaluated the effect of RND and BLD on animal research. The authors' objectives were to determine the proportion of animal studies presented at a national academic emergency medicine meeting that utilize randomization, blinding, or both; and to determine whether failure to employ these techniques changes the likelihood of observing a difference between treatment groups. METHODS: Two trained researchers reviewed abstracts presented at the 1997-2001 Society for Academic Emergency Medicine (SAEM) annual meetings using a standard data collection sheet. Studies that used an animal or cell line, compared two or more study groups, and measured an effect caused by the intervention or drugs were included. Studies were classified as randomized (RND+) if any part of the experiment involved random assignment of subjects to treatment groups, blinded (BLD+) if any assessment of the outcome was made by an investigator blinded to treatment group, and outcome-positive (Outcome+) if any difference between the study groups met the author's definition of significant. Following the initial review, differences in classification were resolved by consensus. The association between outcome and study methodology (RND, BLD or both) was measured using odds ratios (ORs) with 95% confidence intervals (95% CIs). RESULTS: A total of 2,592 studies were published as abstracts. Three hundred eighty-nine were animal studies, and 290 of these studies had two or more study groups. RND- and BLD- studies were more likely to be Outcome+ than RND+ or BLD+ studies (OR = 3.4; 95% CI = 1.7 to 6.9 and OR = 3.2; 95% CI = 1.3 to 7.7, respectively). When studies that used both RND and BND were compared with studies that used neither, the OR for a positive study was 5.2 (95% CI = 2.0 to 13.5). CONCLUSIONS: These results suggest that animal studies that do not utilize RND and BLD are more likely to report a difference between study groups than studies that employ these methods. PMID- 12782534 TI - Magnesium-aluminum hydroxide suspension for the treatment of dermal capsaicin exposures. AB - OBJECTIVES: To determine whether magnesium-aluminum-hydroxide-simethicone suspension (MgAl) is an effective treatment for dermal capsaicin exposures. METHODS: The authors performed a double-blind, randomized, controlled, pilot study comparing the effect of MgAl with that of saline in the treatment of dermal capsaicin exposures. Ten volunteers were sprayed with a commercial defensive spray containing 10% capsaicin on the flexor surface of both forearms. A dressing embedded with MgAl (Maalox) suspension was randomly applied to one arm and a saline-embedded dressing was applied to the other arm. Pain was assessed on a 10 cm visual analog scale at 0, 10, 20, 30, 60, 90, and 120 minutes. RESULTS: Mean pain scores were significantly lower in the MgAl group as compared with the saline (S) group at 10, 20, and 30 minutes. Differences in pain scores were not statistically significant at times 60, 90, and 120 minutes. CONCLUSIONS: During the initial 30 minutes of treatment, there was a statistically significant decrease in pain scores with MgAl as compared with saline treatments. Although the difference in means may have questionable clinical significance, MgAl is cheap and readily available, and has minimal side effects. Thus, MgAl may be an appropriate treatment for dermal capsaicin exposure. PMID- 12782536 TI - "No free lunch" meets the academic journals. PMID- 12782537 TI - Late seizure following ingestion of Vicks VapoRub. PMID- 12782538 TI - To wash or not to wash? That is the question. PMID- 12782539 TI - VO2MAX, blood doping, and erythropoietin. PMID- 12782540 TI - Super athletes or gene cheats? PMID- 12782541 TI - Ottawa ankle rules for the injured ankle. PMID- 12782542 TI - Chiropractic spinal manipulation for back pain. PMID- 12782545 TI - Four weeks of androstenedione supplementation diminishes the treatment response in middle aged men. AB - OBJECTIVES: To examine baseline hormonal concentrations and the pharmacokinetic response on day 0 and day 28 of 28 days of androstenedione supplementation. METHODS: Eight men (mean (SD) age 44.1 (3.0) years (range 40-48), weight 76.3 (9.4) kg, and percentage body fat 20.6 (6.7)) participated in a randomised, double blind, cross over, 2 x 28 day placebo controlled study. Subjects were tested on day 0 and 28 days after receiving 200 mg/day oral androstenedione and a placebo treatment with a 28 day washout period between treatments. Serum hormone concentrations were examined at baseline (time 0) and then at 30 minute intervals for 180 minutes to measure day 0 and day 28 pharmacokinetic responses. Analytes included androstenedione, total testosterone, dehydroepiandrosterone sulfate (DHEAS), oestradiol, and sex hormone binding globulin (SHBG). Lipid concentrations, weight, body composition, resting heart rate, and blood pressure were also measured. RESULTS: Analysis of integrated area under the curve (AUC) and time 0 hormonal concentrations by repeated measures multivariate analysis of variance (p<0.05) and Fisher's post hoc analysis showed a significant increase in AUC for serum androstenedione at day 0 (108.3 (27.6) nmol/l) in the supplemented condition as compared with day 28 (43.4 (13.1) nmol/l) and placebo (2.1 (0.8) nmol/l) conditions. No other significant AUC changes were noted. After 28 days of supplementation, DHEAS levels were significantly elevated (p = 0.00002) at time 0 (12.9 (1.3) micro mol/l) compared with placebo (7.0 (0.8) micro mol/l) with a trend (p = 0.08) toward elevation of time 0 androstenedione concentrations (16.4 (7.0) nmol/l) compared with placebo (5.6 (0.4) nmol/l). No changes were found for lipids, resting heart rate, or blood pressure, weight, or percentage body fat. CONCLUSION: Although supplementation with 200 mg/day androstenedione increases AUC for serum androstenedione in the day 0 condition, continued supplementation is characterised by a diminished treatment response, coupled with time 0 increases in testosterone precursors but not testosterone. PMID- 12782544 TI - Acetabular labrum and its tears. AB - Groin pain is a major cause of morbidity in athletes. Only in the last decade have acetabular labrum tears been recognised as a possible diagnosis. Awareness of this condition is important for appropriate management. The basic science and pathological and clinical features of acetabular labrum tears are reviewed, and diagnostic and treatment options are presented. PMID- 12782546 TI - The current status of sports medicine training in United States internal medicine residency programmes. AB - OBJECTIVE: To determine the general status of sports medicine training in internal medicine residency programmes in the United States. METHODS: A cross sectional survey of the programme directors and chief residents of each of the 407 accredited internal medicine programmes listed in the 1999-2000 Graduate Medical Education Directory. RESULTS: The questionnaire was returned by 231 of 404 (57%) programme directors and 233 of 404 (58%) chief residents. A chief and director of the same programme (paired responses) replied from 144 of 404 (36%) programmes surveyed. A formal sports medicine curriculum was reported by 22.1% of programme directors. Programmes with a formal curriculum were 2.9 times more likely to offer any of the sports medicine educational experiences (p<0.0001; Cochran-Mantel-Haenszel). Programmes with block rotations were more likely to include all of the educational experiences surveyed than those without (p<0.002 for each; chi(2) test). A total of 162 programmes included sports medicine as part of other rotations. Most programmes only included sports medicine as part of other rotations: 44.6% (103/231) of all programmes and 63.6% (103/162) of programmes with sports medicine as part of other rotations. Some 29.9% (69/231) of directors reported having an elective, and 3.9% (9/231) reported a required rotation. Almost a quarter (21.7%; 50/231) of directors reported that their residents received no clinical experience in sports medicine. CONCLUSIONS: Little attention is given to the subject of sports medicine when internal medicine residency curricula are developed in the United States. Thus only a small percentage of American internal medicine residency programmes provide significant training in sports medicine. PMID- 12782543 TI - Limits to the measurement of habitual physical activity by questionnaires. AB - Despite extensive use over 40 years, physical activity questionnaires still show limited reliability and validity. Measurements have value in indicating conditions where an increase in physical activity would be beneficial and in monitoring changes in population activity. However, attempts at detailed interpretation in terms of exercise dosage and the extent of resulting health benefits seem premature. Such usage may become possible through the development of standardised instruments that will record the low intensity activities typical of sedentary societies, and will ascribe consistent biological meaning to terms such as light, moderate, and heavy exercise. PMID- 12782547 TI - Football incident analysis: a new video based method to describe injury mechanisms in professional football. AB - OBJECTIVES: To develop and test a new video based method for match analysis that combines football specific and medical information to achieve a better understanding of the injury mechanisms and events leading up to high risk situations. METHODS: Football incident analysis (FIA) is a video based method describing incidents that may result in an injury using 19 variables and categories modified from match analysis. Videos from 35 of 76 (46%) official Norwegian under 21 matches played from 1994 to 1998 were analysed. Two football experts classified each incident on the basis of predetermined criteria, and their results were compared using interobserver and intraobserver reliability tests. RESULTS: kappa correlation coefficients for interobserver and intraobserver agreement were very good for 63% and 95% and good for 37% and 5% of the variables respectively. Fifty two incidents were recorded (1.6 incidents per team per match or 94 per 1000 player hours), and 16 (31%) led to injuries (0.5 injuries per match or 29 injuries per 1000 player hours). FIA results showed that 28 incidents occurred while attacking in midfield zone 2 or the attacking zone, and 24 took place while defending in the defensive zone or midfield zone 1. Midfielders were exposed in 67% of the incidents, mainly in breakdown attacks or during long attacks by the opposing team. Of the 28 incidents during offence, only one was classified as having great potential to score a goal. Most incidents (70%) were the result of tackling duels both in the offensive and defensive playing phases. Of the 21 offensive incidents resulting from tackling duels, in 19 cases the exposed player was unaware of the tackling (passive duellist). CONCLUSIONS: This study shows that football incident analysis is a potentially valuable tool for understanding the events leading up to injuries in football. PMID- 12782548 TI - The Football Association Medical Research Programme: an audit of injuries in professional football: an analysis of ankle sprains. AB - AIM: To conduct a detailed analysis of ankle sprains sustained in English professional football over two competitive seasons. METHODS: Club medical staff at 91 professional football clubs annotated player injuries. A specific injury audit questionnaire was used together with a weekly form that documented each club's current injury status. RESULTS: Completed injury records for the two competitive seasons were obtained from 87% and 76% of the participating clubs. Ankle ligament sprains accounted for 11% of the total injuries over the two seasons, with over three quarters (77%) of sprains involving the lateral ligament complex. A total of 12 138 days and 2033 matches were missed because of ankle sprains. More sprains were caused by contact mechanisms than non-contact mechanisms (59% v 39%) except in goalkeepers who sustained more non-contact sprains (21% v 79%, p<0.01). Ankle sprains were most often observed during tackles (54%). More ankle sprains were sustained in matches than in training (66% v 33%), with nearly half (48%) observed during the last third of each half of matches. A total of 44% of sprains occurred during the first three months of the season. A high number of players (32%) who sustained ankle sprains were wearing some form of external support. The recurrence rate for ankle sprains was 9% (see methodology for definition of reinjury). CONCLUSION: Ankle ligament sprains are common in football usually involving the lateral ligament complex. The high rate of occurrence and recurrence indicates that prevention is of paramount importance. PMID- 12782549 TI - A prospective study of running injuries: the Vancouver Sun Run "In Training" clinics. AB - OBJECTIVES: Seventeen running training clinics were investigated to determine the number of injuries that occur in a running programme designed to minimise the injury rate for athletes training for a 10 km race. The relative contributions of factors associated with injury were also reported. METHODS: A total of 844 primarily recreational runners were surveyed in three trials on the 4th, 8th, and 12th week of the 13 week programme of the "In Training" running clinics. Participants were classified as injured if they experienced at least a grade 1 injury-that is, pain only after running. Logistic regression modelling and odds ratio calculation were performed for each sex using the following predictor variables: age, body mass index (BMI), previous aerobic activity, running frequency, predominant running surface, arch height, running shoe age, and concurrent cross training. RESULTS: Age played an important part in injury in women: being over 50 years old was a risk factor for overall injury, and being less than 31 years was protective against new injury. Running only one day a week showed a non-significant trend for injury risk in men and was a significant risk factor in women and overall injury. A BMI of > 26 kg/m(2) was reported as protective for men. Running shoe age also significantly contributed to the injury model. Half of the participants who reported an injury had had a previous injury; 42% of these reported that they were not completely rehabilitated on starting the 13 week training programme. An injury rate of 29.5% was recorded across all training clinics surveyed. The knee was the most commonly injured site. CONCLUSIONS: Although age, BMI, running frequency (days a week), and running shoe age were associated with injury, these results do not take into account an adequate measure of exposure time to injury, running experience, or previous injury and should thus be viewed accordingly. In addition, the reason for the discrepancy in injury rate between these 17 clinics requires further study. PMID- 12782551 TI - Cricket injuries: a longitudinal study of the nature of injuries to South African cricketers. AB - OBJECTIVE: To determine the incidence and nature of injuries sustained by elite cricketers during a three season period in order to identify possible injury patterns. METHODS: Thirty six physiotherapists and 13 doctors working with 11 provincial and the South African national teams completed a questionnaire for each cricketer who presented with an injury during each season to determine anatomical site of injury, month of injury during the season, diagnosis, mechanism of injury, whether it was a recurrence of a previous injury, whether the injury had recurred again during the season, and biographical data. RESULTS: A total of 436 cricketers sustained 812 injuries. Bowling (41.3%), fielding and wicketkeeping (28.6%), and batting (17.1%) accounted for most of the injuries. The lower limbs (49.8%), upper limbs (23.3%), and back and trunk (22.8%) were most commonly injured. The injuries occurred primarily during first class matches (27.0%), limited overs matches (26.9%), and practices (26.8%) during the early part of the season. Acute injuries made up 64.8% of the injuries. The younger players (up to 24 years) sustained 57% of the first time injuries, and the players over 24 years of age sustained 58.7% of the injuries that recurred from a previous season. The injuries were mainly soft tissue injuries predominantly to muscle (41.0%), joint (22.2%), tendon (13.2%), and ligament (6.2%). The primary mechanism of injury was the delivery and follow through of the fast bowler (25.6%), overuse (18.3%), and fielding (21.4%). CONCLUSION: The results indicate a pattern of cause of injury, with the young fast bowler most likely to sustain an acute injury to the soft tissues of the lower limb while participating in matches and practices during the early part of the season. PMID- 12782550 TI - Energy demands during a judo match and recovery. AB - OBJECTIVE: To assess energy demand during a judo match and the kinetics of recovery by measuring the metabolites of the oxypurine cascade, lipolytic activity, and glycolytic pathway. METHODS: Venous blood samples were taken from 16 national judoists (mean (SEM) age 18.4 (1.6) years), before (T(1)) and three minutes (T(2)), one hour (T(3)), and 24 hours (T(4)) after a match. A seven day diet record was used to evaluate nutrient intake. RESULTS: Nutrient analysis indicated that these athletes followed a low carbohydrate diet. Plasma lactate concentration had increased to 12.3 (1.8) mmol/l at the end of the match. An increase in the levels of extracellular markers of muscle adenine nucleotide catabolism, urea, and creatinine was observed at T(2), while uric acid levels remained unchanged. High concentrations of urea persisted for 24 hours during the recovery period. Ammonia, hypoxanthine, xanthine, and creatinine returned to control levels within the 24 hour recovery period. Uric acid concentrations rose from T(3) and had not returned to baseline 24 hours after the match. The levels of triglycerides, glycerol, and free fatty acids had increased significantly (p<0.05) after the match (T(2)) but returned to baseline values within 24 hours. Concentrations of high density lipoprotein cholesterol and total cholesterol were significantly increased after the match. CONCLUSIONS: These results show that a judo match induces both protein and lipid metabolism. Carbohydrate availability, training adaptation, and metabolic stress may explain the requirement for these types of metabolism. PMID- 12782552 TI - Heart rate responses of women aged 23-67 years during competitive orienteering. AB - OBJECTIVES: To compare the heart rate responses of women orienteers of different standards and to assess any relation between heart rate responses and age. METHODS: Eighteen competitive women orienteers completed the study. They were divided into two groups: eight national standard orienteers (ages 23-67 years); 10 club standard orienteers (ages 24-67 years). Each participant had her heart rate monitored during a race recognised by the British Orienteering Federation. Peak heart rate (HR(PEAK)), mean heart rate (HR(MEAN)), standard deviation of her heart rate during each orienteering race (HR(SD)), and mean change in heart rate at each control point (DeltaHR(CONTROL)) were identified. The data were analysed using analysis of covariance with age as a covariate. RESULTS: National standard orienteers displayed a lower within orienteering race standard deviation in heart rate (6 (2) v 12 (2) beats/min, p<0.001) and a lower DeltaHR(CONTROL) (5 (1) v 17 (4) beats/min, p<0.001). The mean heart rate during competition was higher in the national standard group (170 (11) v 158 (11) beats/min, p = 0.025). The HR(MEAN) for the national and club standard groups were 99 (8)% and 88 (9)% of their age predicted maximum heart rate (220-age) respectively. All orienteers aged >55 years (n = 4) recorded HR(MEAN) greater than their age predicted maximum. CONCLUSIONS: The heart rate responses indicate that national and club standard women orienteers of all ages participate in the sport at a vigorous intensity. The higher DeltaHR(CONTROL) of club standard orienteers is probably due to failing to plan ahead before arriving at the controls and this, coupled with slowing down to navigate or relocate when lost, produced a higher HR(SD). PMID- 12782553 TI - Peroneus longus stretch reflex amplitude increases after ankle brace application. AB - BACKGROUND: The use of external ankle support is widespread throughout sports medicine. However, the application of ankle bracing to a healthy ankle over a long period has been scrutinised because of possible neuromuscular adaptations resulting in diminished dynamic support offered by the peroneus longus. OBJECTIVE: To investigate the immediate and chronic effects of ankle brace application on the amplitude of peroneus longus stretch reflex. METHODS: Twenty physically active college students (mean (SD) age 23.6 (1.7) years, height 168.7 (8.4) cm, and mass 69.9 (12.0) kg) who had been free from lower extremity pathology for the 12 months preceding the study served as subjects. None had been involved in a strength training or conditioning programme in the six months preceding the study. A 3 x 3 x 2 (test condition x treatment condition x time) design with repeated measures on the first and third factor was used. The peroneus longus stretch reflex (% of maximum amplitude) during sudden foot inversion was evaluated under three ankle brace conditions (control, lace up, and semi-rigid) before and after eight weeks of ankle brace use. RESULTS: A 3 x 3 x 2 repeated measures analysis of variance showed that peroneus longus stretch reflex amplitude increased immediately after application of a lace up brace (67.1 (4.4)) compared with the semi-rigid (57.9 (4.3)) and control (59.0 (5.2)) conditions (p<0.05). Peroneus longus stretch reflex also increased after eight weeks of use of the semi-rigid brace compared with the lace up and control conditions (p<0.05). CONCLUSIONS: Initial application of a lace up style ankle brace and chronic use of a semi-rigid brace facilitates the amplitude of the peroneus longus stretch reflex. It appears that initial and long term ankle brace use does not diminish the magnitude of this stretch reflex in the healthy ankle. PMID- 12782555 TI - Day to day variability in the transferrin receptor/ferritin index in female athletes. AB - OBJECTIVE: To determine the within subject (day to day) variability for variables reflecting iron status (concentrations of ferritin and soluble transferrin receptor (sTfR), and the sTfR/logFerr index) in female athletes during a training camp, and in female control subjects. RESULTS: The error for ferritin concentration was nearly twice as high in athletes as in controls, that for sTfR concentration was identical in the two groups, and that for the sTfR/logFerr index was about 50% higher in athletes than in controls. CONCLUSIONS: The within subject, day to day error for the sTfR/logFerr index computed from data recorded for untrained subjects cannot serve as a reference value for training athletes. When the sTfR/logFerr index is used to monitor iron stores in athletes, an error value of 0.20 should be used, because determination of the index after a few days of rest may not be feasible. PMID- 12782554 TI - A pilot study of the prevalence of lumbar disc degeneration in elite athletes with lower back pain at the Sydney 2000 Olympic Games. AB - OBJECTIVES: To observe the prevalence of lumbar intervertebral disc degeneration in elite athletes as compared with published literature of changes seen in non athletes-that is, normal population. METHODS: The lumbar spines of 31 Olympic athletes who presented to the Olympic Polyclinic with low back pain and/or sciatica were examined using magnetic resonance imaging. Three criteria were looked at: (a) the loss of disc signal intensity; (b) the loss of disc height; (c) the presence of disc displacement. The results were then recorded and correlated with the lumbar levels. RESULTS: The disc signal intensity was progressively reduced the more caudal the disc space. It was most common at the L5/S1 level, and, of the abnormal group, 36% (n = 11) showed the most degenerative change. Disc height reduction was also found to be most common at the L5/S1 level. However, the most common height reduction was only mild. A similar trend of increased prevalence of disc herniation was noted with more caudal levels. At the L5/S1 level, 58% were found to have an element of disc displacement, most of which were disc bulges. Compared with changes seen in the normal population (non-athletes) as described in the literature, disc degeneration defined by the above criteria was found to be significantly more severe in these Olympic athletes. CONCLUSIONS: Although the study was limited, the results suggest that elite athletes have a greater prevalence and greater degree of lumbar disc degeneration than the normal population. A more detailed follow up study should be considered to investigate which particular training activities have the most impact on the lumbar spine, and how to modify training methods so as to avoid the long term sequelae of degenerative disc disease of the lumbar spine. PMID- 12782556 TI - Bilateral radial head fractures in a martial arts athlete. AB - Isolated radial head fractures, which are rare, comprise about 2% of all fractures around the elbow. A case is reported of bilateral radial head fracture in an international female athlete. Early recognition and aggressive rehabilitation is essential if an abrupt end to a promising sporting career is to be prevented. PMID- 12782557 TI - Lacrosse stick entrapment injury to the thumb. AB - A case of injury to the left thumb following an errant stick check, and subsequent entrapment of the digit in the open sidewall of a lacrosse stick, is presented. A circumferential laceration, severe swelling, and bruising to the proximal phalanx resulted. This case report emphasises the need to limit the dimensions of openings in the sidewalls of lacrosse sticks to prevent the occurrence of this and other preventable injuries. PMID- 12782558 TI - Relation between the occurrence of bony union and outcome for fractures of the lateral process of the talus: a case report and analysis of published reports. AB - Fractures of the lateral process of the talus are uncommon, but the incidence has probably risen because of the increased popularity of snowboarding, an increased awareness of doctors, and the ready availability of computed tomography scanning. One case report and a review of the literature are provided. Aggressive management rarely resulted in non-union (7%) and was always associated with a good outcome (100%), even if the fracture fragments were excised initially. The relations between method of treatment, bony union, and outcome were significant (Fisher's exact test, p<0.01). PMID- 12782559 TI - The forgotten Barcroft/Edholm reflex: potential role in exercise associated collapse. PMID- 12782560 TI - Comparative body fat assessment in elite footballers. PMID- 12782561 TI - Lumbopelvic mechanics. PMID- 12782562 TI - Understanding tendinopathies. PMID- 12782563 TI - Muscle dysmorphia in weightlifters. PMID- 12782564 TI - New faculty of sport and exercise medicine in Ireland. PMID- 12782565 TI - Intravascular ultrasound analysis of infarct-related and non-infarct-related arteries in patients who presented with an acute myocardial infarction. AB - BACKGROUND: Previous studies have reported diffuse destabilization of atherosclerotic plaques in acute myocardial infarction (AMI). METHODS AND RESULTS: We used intravascular ultrasound (IVUS) to assess 78 coronary arteries (38 infarct-related arteries [IRAs] with culprit and nonculprit lesions and 40 non-IRAs) from 38 consecutive AMI patients. IVUS analysis included qualitative and quantitative measurements of reference and lesion external elastic membrane (EEM), lumen, and plaque plus media (P&M) area. Positive remodeling was defined as lesion/mean reference EEM >1.0. Culprit lesions were identified by a combination of ECG, wall motion abnormalities (ventriculogram or echocardiogram), scintigraphic perfusion defects, and coronary angiogram. Culprit lesions contained more thrombus (23.7% versus 3.4% in nonculprit IRA plaques and 3.1% in non-IRA plaques; P=0.0011). Culprit lesions were predominantly hypoechoic (63.2% versus 37.9% of nonculprit IRA plaques and 28.1% of non-IRA plaques; P=0.0022). Culprit lesions were longer (17.5+/-10.1, 9.8+/-4.0, and 10.3+/-5.7 mm, respectively; P<0.0001), had larger EEM area (15.0+/-6.0, 11.5+/-5.7, and 12.6+/ 5.6 mm2, respectively; P=0.0353) and P&M area (13.0+/-6.0, 7.5+/-3.7, 9.3+/-4.3 mm2, respectively; P<0.0001), smaller lumens (2.0+/-0.9, 4.1+/-3.1, and 3.4+/-2.5 mm2, respectively; P=0.0009), and more positive remodeling (79.4%, 59.0%, and 50.8%, respectively; P=0.0155). The frequency of plaque rupture/dissection was greater in culprit, nonculprit IRA, and non-IRA plaques in AMI patients than in a control group of chronic stable angina patients with multivessel IVUS imaging. CONCLUSIONS: Culprit plaques have more markers of instability (thrombus, positive remodeling, and large plaque mass); however, these markers of instability are not typically found elsewhere. This suggests that the vascular event in AMI patients is determined by local pre-event lesion morphologies. PMID- 12782566 TI - Adverse effect of ventricular pacing on heart failure and atrial fibrillation among patients with normal baseline QRS duration in a clinical trial of pacemaker therapy for sinus node dysfunction. AB - BACKGROUND: Dual-chamber (DDDR) pacing preserves AV synchrony and may reduce heart failure (HF) and atrial fibrillation (AF) compared with ventricular (VVIR) pacing in sinus node dysfunction (SND). However, DDDR pacing often results in prolonged QRS durations (QRSd) as the result of right ventricular stimulation, and ventricular desynchronization may result. The effect of pacing-induced ventricular desynchronization in patients with normal baseline QRSd is unknown. METHODS AND RESULTS: Baseline QRSd was obtained from 12-lead ECGs before pacemaker implantation in MOST, a 2010-patient, 6-year, randomized trial of DDDR versus VVIR pacing in SND. Cumulative percent ventricular paced (Cum%VP) was determined from stored pacemaker data. Baseline QRSd <120 ms was observed in 1339 patients (707 DDDR, 632 VVIR). Cum%VP was greater in DDDR versus VVIR (90% versus 58%, P=0.001). Cox models demonstrated that the time-dependent covariate Cum%VP was a strong predictor of HF hospitalization in DDDR (hazard ratio [HR], 2.99 [95% CI, 1.15 to 7.75] for Cum%VP >40%) and VVIR (HR 2.56 [95% CI, 1.48 to 4.43] for Cum%VP >80%). The risk of AF increased linearly with Cum%VP from 0% to 85% in both groups (DDDR, HR 1.36 [95% CI, 1.09, 1.69]; VVIR, HR 1.21 [95% CI 1.02, 1.43], for each 25% increase in Cum%VP). Model results were unaffected by adjustment for known baseline predictors of HF hospitalization and AF. CONCLUSIONS: Ventricular desynchronization imposed by ventricular pacing even when AV synchrony is preserved increases the risk of HF hospitalization and AF in SND with normal baseline QRSd. PMID- 12782567 TI - Transgenic mice overexpressing mutant PRKAG2 define the cause of Wolff-Parkinson White syndrome in glycogen storage cardiomyopathy. AB - BACKGROUND: Mutations in the gamma2 subunit (PRKAG2) of AMP-activated protein kinase produce an unusual human cardiomyopathy characterized by ventricular hypertrophy and electrophysiological abnormalities: Wolff-Parkinson-White syndrome (WPW) and progressive degenerative conduction system disease. Pathological examinations of affected human hearts reveal vacuoles containing amylopectin, a glycogen-related substance. METHODS AND RESULTS: To elucidate the mechanism by which PRKAG2 mutations produce hypertrophy with electrophysiological abnormalities, we constructed transgenic mice overexpressing the PRKAG2 cDNA with or without a missense N488I human mutation. Transgenic mutant mice showed elevated AMP-activated protein kinase activity, accumulated large amounts of cardiac glycogen (30-fold above normal), developed dramatic left ventricular hypertrophy, and exhibited ventricular preexcitation and sinus node dysfunction. Electrophysiological testing demonstrated alternative atrioventricular conduction pathways consistent with WPW. Cardiac histopathology revealed that the annulus fibrosis, which normally insulates the ventricles from inappropriate excitation by the atria, was disrupted by glycogen-filled myocytes. These anomalous microscopic atrioventricular connections, rather than morphologically distinct bypass tracts, appeared to provide the anatomic substrate for ventricular preexcitation. CONCLUSIONS: Our data establish PRKAG2 mutations as a glycogen storage cardiomyopathy, provide an anatomic explanation for electrophysiological findings, and implicate disruption of the annulus fibrosis by glycogen-engorged myocytes as the cause of preexcitation in Pompe, Danon, and other glycogen storage diseases. PMID- 12782568 TI - Targeting of mitogen-activated protein kinases and phosphatidylinositol 3 kinase inhibits hepatocyte growth factor/scatter factor-induced angiogenesis. AB - BACKGROUND: Hepatocyte growth factor/scatter factor (HGF/SF) can sufficiently and independently induce pathophysiological angiogenesis. However, the treatment strategies have mostly been unsuccessful. The present study is the first to evaluate the possible targeting of downstream signals for the inhibition of HGF/SF-induced angiogenesis. METHODS AND RESULTS: In a multichannel scratch assay with human endothelial cells (ECs), HGF/SF induced a strong and prolonged activation of MAPK and cell proliferation that was inhibited by PD98059 and LY294002/wortmannin, selective inhibitors of MAPK and PI3K signaling modules, respectively. Western blotting demonstrated a temporal relation between the activation of the two pathways. Chemical inhibition of the PI3K and MAPK signals inhibited HGF/SF-induced chemoinvasion of ECs in vitro and blocked the HGF/SF induced neovascularization into a polymer scaffold in vivo, as quantified by vessel counts and the clearance of radioactive 133Xe. CONCLUSIONS: These data indicate that MEK and PI3K inhibitors represent a promising approach to the clinical management of pathological conditions characterized by overt HGF/SF induced angiogenesis. PMID- 12782569 TI - New three-dimensional echocardiographic system using digital radiofrequency data- visualization and quantitative analysis of aortic valve dynamics with high resolution: methods, feasibility, and initial clinical experience. AB - BACKGROUND: Common 3D systems have only limited spatial and temporal resolution (frame rate of 25 Hz). Thin structures such as cardiac valves are not imaged exactly; rapid movement patterns cannot be precisely recorded. The objective of the present project was to achieve radiofrequency (RF) data transmission to the 3D workstation to improve image resolution. METHODS AND RESULTS: A commercially available echocardiographic system (5-MHz transesophageal echocardiography probe) with an integrated raw data interface enables transmission of RF data (up to 40 megabytes per second). A 3D data set may contain up to 3 gigabytes, so that all of the high-resolution ultrasound information of the 2D image is available. Frame rates of up to 168 Hz result in temporal resolution 6 times that of standard 3D systems. The applicability of the system and the image quality were tested in 10 patients. The structure of the aortic valve and the dynamic changes were depicted by volume rendering. The changes in the orifice areas were measured in frame-by frame planimetry. The mean number of frames recorded per cardiac cycle was 122+/ 16. The improved structural resolution enabled a detailed imaging of the morphology of the aortic cusps. The rapid systolic movement patterns were recorded with up to 51 frames. The high number of frames enabled creation of precise area-time diagrams. Thus, the individual phases of aortic valve movement (rapid opening, slow valve closing, and rapid valve closing) could be analyzed quantitatively. CONCLUSIONS: A 3D system based on RF data enables high-resolution imaging of cardiac movement patterns. This offers new perspectives for qualitative and quantitative analyses, especially of cardiac valves. PMID- 12782570 TI - Locus for atrial fibrillation maps to chromosome 6q14-16. AB - BACKGROUND: Atrial fibrillation (AF), the most common clinical arrhythmia, is a major cause of morbidity and mortality. Although AF is often associated with other cardiovascular conditions, many patients present without an obvious etiology. Inherited forms of AF exist, but the causative gene has been defined only in a single family. We have identified a large family (family FAF-1) in which AF segregates as a Mendelian trait. METHODS AND RESULTS: Thirty-four family members were evaluated by 12-lead ECG, echocardiogram, 24-hour Holter monitoring, and laboratory studies. Individuals with electrocardiographically documented AF were defined as affected. Subjects were considered unaffected if they were >60 years of age, had no personal history of AF, and had no offspring with a history of AF. DNA was extracted and genotypic analyses were performed using polymorphic microsatellite markers. Evidence of linkage was obtained on chromosome 6, with a peak 2-point logarithm of the odds (LOD) score of 3.63 (theta=0) at the marker D6S1021. A maximal multipoint LOD score of 4.9 was obtained between D6S286 and D6S1021, indicating odds of approximately 100 000:1 in favor of this interval as the location of the gene defect responsible for AF in this family. The LOD scores were robust to changes in penetrance and allele frequency. Haplotype analyses further supported this minimal genetic interval. CONCLUSIONS: We have mapped a novel locus for AF to chromosome 6q14-16. The identification of the causative gene in this interval will be an important step in understanding the fundamental mechanisms of AF. PMID- 12782571 TI - Tumor cell senescence in cancer treatment. AB - Cell senescence is broadly defined as the physiological program of terminal growth arrest, which can be triggered by alterations of telomeres or by different forms of stress. Neoplastic transformation involves events that inhibit the program of senescence, and tumor cells were believed until recently to have lost the ability to senesce. It has now become apparent, however, that tumor cells can be readily induced to undergo senescence by genetic manipulations or by treatment with chemotherapeutic drugs, radiation, or differentiating agents. Treatment induced senescence, which has both similarities with, and differences from, replicative senescence of normal cells, was shown to be one of the key determinants of tumor response to therapy in vitro and in vivo. Although senescent cells do not proliferate, they remain metabolically active and produce secreted proteins with both tumor-suppressing and tumor-promoting activities. Expression of tumor-promoting factors by senescent cells is mediated, at least in part, by senescence-associated cyclin-dependent kinase inhibitors such as p21(Waf1/Cip1/Sdi1). Clinical and preclinical studies indicate that expression of different biological classes of senescence-associated growth-regulatory genes in tumor cells has significant prognostic implications. Elucidation of the genes and regulatory mechanisms that determine different aspects of tumor senescence makes it possible to design new therapeutic approaches to improving the efficacy and to decreasing the side effects of cancer therapy. PMID- 12782572 TI - PRL-3 and PRL-1 promote cell migration, invasion, and metastasis. AB - We demonstrate here that Chinese hamster ovary cells stably expressing PRL-3, a M(r) 20000 prenylated protein tyrosine phosphatase, or its relative, PRL-1, exhibit enhanced motility and invasive activity. A catalytically inactive PRL-3 mutant has significantly reduced migration-promoting activity. We observe that PRL-3 is associated with diverse membrane structures involved in cell movement. Furthermore, we show that PRL-3- and -1-expressing cells, but not control cells, induce metastatic tumor formation in mice. Thus, our results deliver the first evidence for a causative role of PRL-3 and -1 in promoting cell motility, invasion activity, and metastasis. PMID- 12782573 TI - Magnetic resonance molecular imaging of the HER-2/neu receptor. AB - The HER-2/neu receptor is a member of the epidermal growth factor family and is amplified in multiple cancers. It is under intense investigation both as a prognostic marker and for therapy, using monoclonal antibodies targeted against the receptor. We have developed a novel two-component gadolinium-based MR contrast agent to image the HER-2/neu receptor. Positive T1 contrast in MR images was generated by the specific binding of avidin-gadolinium complexes to tumor cells prelabeled with a biotinylated anti-HER-2/neu antibody. Significant intensity enhancement was observed in HER-2/neu-expressing cell lines and in vivo in a breast cancer model. Potential applications of this approach may include determination of the HER-2/neu status for prognosis and for selecting tumors for monoclonal antibody therapy. PMID- 12782574 TI - Development of rhabdomyosarcoma in HER-2/neu transgenic p53 mutant mice. AB - Rhabdomyosarcomas derive from the skeletal muscle lineage and harbor a variety of genetic and molecular lesions. However, it is not clear which molecular alterations have a pathogenetic role. We show that activation of the HER-2/neu oncogene coupled with inactivation of the oncosuppressor gene p53 causes rhabdomyosarcoma in mice. At the age of 11-21 weeks, all male mice carrying both genetic lesions developed embryonal rhabdomyosarcomas expressing desmin, myosin, and insulin-like growth factor-II, in the genitourinary tract. Our findings led to the hypothesis that the interaction between HER family genes and the p53 pathway might be involved in the origin of human rhabdomyosarcoma. PMID- 12782575 TI - Mouse embryos cloned from brain tumors. AB - Cancer cells escape from growth control by accumulating genetic and epigenetic alterations. In rare instances, epigenetic changes alone are oncogenic. Furthermore, agents that modify DNA methylation or chromatin structure can restore a normal phenotype to cells harboring oncogenic mutations. However, it is unclear to what extent epigenetic reprogramming can reverse oncogenesis. Using somatic nuclear transfer, we show that medulloblastomas arising in Ptc1+/- mice can direct preimplantation development. Additionally, blastocysts derived from medulloblastoma nuclei form postimplantation embryos with typical cell layers. Thus, tumor cells can be epigenetically reprogrammed into normal cell types. This approach could lead to a general strategy for assessing genetic and epigenetic contributions to tumorigenesis. PMID- 12782576 TI - p73 is effective in p53-null pancreatic cancer cells resistant to wild-type TP53 gene replacement. AB - Novel therapies such as gene therapy are needed for the treatment of pancreatic carcinomas. Here we show that adenovirus-mediated p73 overexpression results in a strong induction of apoptosis, whereas the effect of p53 varies between different cell lines. In particular, p53-negative AsPC-1 cells are resistant to p53 mediated apoptosis. In these cells, only ectopically expressed p73 activates the proapoptotic p53 target P53AIP1, whereas phosphorylation of p53 at Ser-46, shown to regulate transcriptional activation of P53AIP1, is missing. Our findings support the use of p73 as an anticancer drug in p53-null pancreatic cancer cells that are resistant to wild-type TP53 gene replacement. PMID- 12782577 TI - Analysis of the phosphatidylinositol 3'-kinase signaling pathway in glioblastoma patients in vivo. AB - Deregulated signaling through the phosphatidylinositol 3'-kinase (PI3K) pathway is common in many types of cancer, including glioblastoma. Dissecting the molecular events associated with activation of this pathway in glioblastoma patients in vivo presents an important challenge that has implications for the development and clinical testing of PI3K pathway inhibitors. Using an immunohistochemical analysis applied to a tissue microarray, we performed hierarchical clustering and multidimensional scaling, as well as univariate and multivariate analyses, to dissect the PI3K pathway in vivo. We demonstrate that loss of the tumor suppressor protein PTEN, which antagonizes PI3K pathway activation, is highly correlated with activation of the main PI3K effector Akt in vivo. We also show that Akt activation is significantly correlated with phosphorylation of mammalian target of rapamycin (mTOR), the family of forkhead transcription factors (FOXO1, FOXO3a, and FOXO4), and S6, which are thought to promote its effects. Expression of the mutant epidermal growth factor receptor vIII is also tightly correlated with phosphorylation of these effectors, demonstrating an additional route to PI3K pathway activation in glioblastomas in vivo. These results provide the first dissection of the PI3K pathway in glioblastoma in vivo and suggest an approach to stratifying patients for targeted kinase inhibitor therapy. PMID- 12782578 TI - Identification of novel genetic loci contributing to 12-O-tetradecanoylphorbol-13 acetate skin tumor promotion susceptibility in DBA/2 and C57BL/6 mice. AB - Genetic differences in susceptibility to two-stage skin carcinogenesis have been known for many years. Studies of genetic crosses of sensitive DBA/2 with resistant C57BL/6 mice suggested that multiple autosomal genes determine the sensitivity of these mice to 12-O-tetradecanoylphorbol-13-acetate (TPA) skin tumor promotion. Previous studies mapped one promotion susceptibility locus, Psl1, to distal chromosome 9. Analysis of TPA promotion susceptibility in (C57BL/6 x DBA/2)F(1) x C57BL/6 mice and B x D recombinant inbred mouse strains suggested tentative associations of promotion susceptibility with several other chromosomal regions. To confirm these associations (C57BL/6 x BxD27)F(2) mice analyzed for TPA promotion susceptibility were genotyped for polymorphic genetic markers mapping to chromosomal regions for which tentative associations had been previously detected. BxD27 mice are sensitive to TPA skin tumor promotion but carry the C57BL/6 allele of Psl1. Because Psl1 does not segregate in this cross, its effect on TPA promotion susceptibility is the same for all mice in the cross. The results of this analysis support the mapping of three novel promotion susceptibility loci to chromosomes 1, 2, and 19. Psl2 maps near D2Mit229 on distal chromosome 2, and inheritance of the dominant DBA/2 allele results in increased sensitivity to TPA. Psl3 maps near D1Mit511 on distal chromosome 1. Interestingly, inheritance of an allele from the resistant C57BL/6 parent results in increased sensitivity to TPA. Psl3 appears to have an additive affect, with heterozygous mice having a stronger response to TPA than mice homozygous for the DBA/2 allele and a weaker response to TPA than mice homozygous for the C57BL/6 allele. Psl4 maps near D19Mit38 on distal chromosome 19 and inheritance of the dominant C57BL/6 allele results in decreased TPA sensitivity. Analysis of the combined effects of these loci on TPA promotion susceptibility indicates that they contribute independently to the overall sensitivity to TPA. PMID- 12782579 TI - Human enzymes involved in the metabolic activation of the environmental contaminant 3-nitrobenzanthrone: evidence for reductive activation by human NADPH:cytochrome p450 reductase. AB - Determining the capability of humans to metabolize the suspected carcinogen 3 nitrobenzanthrone (3-NBA) and understanding which human enzymes are involved in its activation are important in the assessment of individual susceptibility to this environmental contaminant found in diesel exhaust and ambient air pollution. We compared the ability of eight human hepatic microsomal samples to catalyze DNA adduct formation by 3-NBA. Using two enrichment procedures of the (32)P postlabeling method, nuclease P1 digestion and butanol extraction, we found that all hepatic microsomes were competent to activate 3-NBA. DNA adduct patterns with multiple adducts, qualitatively similar to those found recently in vivo in rats, were observed. Additionally one major DNA adduct generated by human microsomes was detected. The role of specific cytochromes p450 (p450) and NADPH:p450 reductase in the human hepatic microsomal samples in 3-NBA activation was investigated by correlating the p450- and NADPH: p450 reductase-linked catalytic activities in each microsomal sample with the level of DNA adducts formed by the same microsomes. On the basis of this analysis, most of the hepatic microsomal activation of 3-NBA was attributed to NADPH: p450 reductase. Inhibition of DNA adduct formation in human liver microsomes by alpha-lipoic acid, an inhibitor of NADPH: p450 reductase, supported this finding. Using the purified rabbit enzyme and recombinant human NADPH: p450 reductase expressed in Chinese hamster V79 cells, we confirmed the participation of this enzyme in the formation of 3-NBA derived DNA adducts. Moreover, essentially the same DNA adduct pattern found in microsomes was detected in metabolically competent human lymphoblastoid MCL-5 cells. The role of individual human recombinant p450s 1A1, 1A2, 1B1, 2A6, 2B6, 2D6, 2C9, 2E1, and 3A4 and of NADPH: p450 reductase in the metabolic activation of 3-NBA, catalyzing DNA adduct formation, was also examined using microsomes of baculovirus-transfected insect cells containing the recombinant enzymes (Supersomes). DNA adducts were observed in all Supersomes preparations, essentially similar to those found with human hepatic microsomes and in human cells. Of all of the recombinant human p450s, p450 2B6 and -2D6 were the most efficient to activate 3-NBA, followed by p450 1A1 and -1A2. These results demonstrate for the first time the potential of human NADPH: p450 reductase and recombinant p450s to contribute to the metabolic activation of 3-NBA by nitroreduction. PMID- 12782580 TI - Estrogen sulfotransferase and steroid sulfatase in human breast carcinoma. AB - Estrogen sulfotransferase (EST; SULT 1E1 or STE gene) sulfonates estrogens to inactive estrogen sulfates, whereas steroid sulfatase (STS) hydrolyzes estrone sulfate to estrone. Both EST and STS have been suggested to play important roles in regulating the in situ production of estrogens in human breast carcinoma tissues. However, the expression of EST has not been examined in breast carcinoma tissues, and the biological significance of EST and STS remains unknown. Therefore, in this study, we examined the expression of EST and STS in 35 specimens of human breast carcinoma tissues using immunohistochemistry, reverse transcription-PCR (RT-PCR), and enzymatic assay. EST and STS immunoreactivity was also correlated with various clinicopathological parameters, including prognosis to examine the biological significance of these enzymes in 113 breast carcinomas. EST and STS immunoreactivity was detected in carcinoma cells and significantly associated with their mRNA levels (P = 0.0027 and 0.0158, respectively), as measured by RT/real-time PCR, and enzymatic activities (P = 0.0005 and 0.0089, respectively) in 35 breast carcinomas. In breast cancer tissues examined by laser capture microdissection/RT-PCR analyses, the mRNA for EST was localized in both carcinoma and intratumoral stromal cells, whereas that of STS was detected only in carcinoma cells. Of the 113 invasive ductal carcinomas examined in this study, EST and STS immunoreactivity was detected in 50 and 84 cases (44.2 and 74.3%), respectively. In these cases, EST immunoreactivity was inversely correlated with tumor size (P = 0.003) or lymph node status (P = 0.0027). In contrast, STS immunoreactivity was significantly correlated with tumor size (P = 0.0047). Moreover, EST immunoreactivity was significantly associated with a decreased risk of recurrence or improved prognosis by both uni (P = 0.0044, and 0.0026, respectively) and multivariate (P = 0.0429 and 0.0149, respectively) analyses. STS immunoreactivity, however, was significantly associated with an increased risk of recurrence (P = 0.0118) and worsened prognosis (P = 0.0325) by univariate analysis. Results from our present study suggest that immunoreactivities for both EST and STS are associated with their mRNA level and enzymatic activity and that EST immunoreactivity is considered to be a potent prognostic factor in human breast carcinoma. PMID- 12782581 TI - Human kallikrein 8, a novel biomarker for ovarian carcinoma. AB - Human kallikrein 8 (hK8; neuropsin) is a serine protease and new member of the hK family. The aim of this study was to examine if hK8 may serve as a novel cancer biomarker. An hk8-ELISA, developed in-house, was used to study the distribution of hK8 in various biological fluids and tissue extracts from healthy individuals and ovarian cancer patients of different stages of the disease (International Federation of Obstetrics and Gynecology II-IV). For ovarian cancer patients, very high levels in ascites fluid were observed (90% block of the metaphase/anaphase transition, and complete inhibition of cell proliferation. The data strongly support the idea that the inhibition of centromere dynamics by Taxol prevents silencing of the mitotic spindle surveillance (checkpoint) mechanism. Because Taxol strongly suppresses microtubule dynamics, the data also indicate that centromere dynamics can be accounted for by microtubule dynamics and may not require significant energetic contributions from microtubule motors. The strict correlation between the degree of suppression of centromere dynamics by Taxol and the degree of mitotic block strongly indicates that the primary mechanism responsible for the mitotic block by Taxol in U2OS cells involves suppression of the polymerization dynamics of kinetochore microtubules. PMID- 12782585 TI - GTI-2040, an antisense agent targeting the small subunit component (R2) of human ribonucleotide reductase, shows potent antitumor activity against a variety of tumors. AB - GTI-2040 is a 20-mer oligonucleotide that is complementary to a coding region in the mRNA of the R2 small subunit component of human ribonucleotide reductase. In vitro studies using a number of human tumor cell lines have demonstrated that GTI 2040 decreases mRNA and protein levels of R2 in a sequence- and target-specific manner. In vivo studies have shown that GTI-2040 significantly inhibits growth of human colon tumors (adenocarcinoma), pancreatic tumors (adenocarcinoma), liver tumors, lung tumors, breast tumors (adenocarcinoma), renal tumors, ovarian tumors (adenocarcinoma), melanoma, brain glioblastoma-astrocytoma, prostatic tumors, and cervical tumors in nude and/or severe combined immunodeficient mice. Antitumor effects were not observed with an oligonucleotide containing four mismatches to the R2 sequence or with a scrambled sequence containing the same base content but not complementary to R2. This suggests that an antisense mechanism is responsible for the in vivo observations. In addition to tumor growth assays, GTI-2040 was tested in a murine model of human lymphoma. Treatment of severe combined immunodeficient mice bearing Burkitt's lymphoma with GTI-2040, but not control oligonucleotides, greatly extended the survival of mice, and survival extended well beyond the treatment period. Finally, GTI-2040 specifically inhibits metastasis of human melanoma cells to the lungs in nude mice. Taken together, the results of these studies indicate that GTI-2040 can act as a selective and specific anticancer agent against a broad range of human tumors. PMID- 12782586 TI - "Vasocrine" formation of tumor cell-lined vascular spaces: implications for rational design of antiangiogenic therapies. AB - Here we report that B16F10 murine melanoma cells mimic endothelial cell behavior and the angiogenic process in vitro and in vivo. Cord formation in vitro by tumor cells is stimulated by hypoxia and vascular endothelial growth factor (VEGF) and inhibited by antibodies against VEGF and the VEGF KDR receptor (VEGF receptor 2). We define regulation of tumor cell-derived vascular space formation by these vasoactive compounds as "vasocrine" stimulation. ICRF 159 (Razoxane; NSC 129943) prevents tumor cell but not endothelial cell cord formation in vitro, and the antiangiogenic drug TNP-470 (NSC 642492) inhibits endothelial but not tumor cell cord formation in vitro. Both drugs inhibit formation of blood-filled vascular spaces in vivo. These results bear on the anticipated action of ICRF 159 in human clinical trials and novel strategies for targeting tumor blood supplies. PMID- 12782587 TI - Modulation of the folate receptor alpha gene by the estrogen receptor: mechanism and implications in tumor targeting. AB - The folate receptor (FR) type alpha is a promising target for diagnostic imaging agents and therapeutic intervention in major subtypes of gynecological malignancies; however, the receptor levels in the tumors are variable and are generally relatively low in estrogen receptor (ER)-positive tumors. Here we report that the FR-alpha gene promoter is repressed in the presence of 17beta estradiol and derepressed by the antiestrogens tamoxifen and ICI 182780 in a promoter-specific and ER-alpha-dependent manner in carcinoma cell lines including HeLa (cervical carcinoma), BG-1 (ovarian carcinoma), and IGROV-1 (ovarian carcinoma). The ligand and ER dose response of the FR-alpha promoter and its time course paralleled those of a classical estrogen response element-mediated effect. Antiestrogens produced an ER-dependent increase of up to 36-fold in the expression of the endogenous FR-alpha gene. Deletion analysis and FR-alpha/SV40 promoter chimeras showed that the ER effect is mediated exclusively within the G/C-rich region in the TATA-less P4 promoter of FR-alpha; electrophoretic mobility shift analysis demonstrated interaction of ER at only one of three G/C rich elements. ER-beta only modestly affected FR-alpha promoter activity but did not diminish the ER-alpha-mediated effects. The ER corepressor, SMRT, enhanced the repression by 17beta-estradiol/ER, but ER coactivators, including SRC family members, did not appreciably impact the ER ligand response. The results suggest that in ER+ tumors, FR-alpha expression is directly and actively suppressed and predict that a brief treatment with antiestrogens will boost FR-alpha expression by passive derepression, enhancing the efficacy of FR-targeted diagnostic and therapeutic applications. They also reveal novel aspects of gene repression by ER. PMID- 12782588 TI - Gene and peptide analyses of newly defined lung cancer antigens recognized by HLA A2402-restricted tumor-specific cytotoxic T lymphocytes. AB - We investigated tumor antigens recognized by HLA-A2402-restricted CTLs established from T cells infiltrating into lung adenocarcinoma. We report here three newly identified tumor antigen genes, including one unreported gene, temporarily referred to as clone 83, and two known genes, BTB domain containing 2 (BTBD2) and hairpin-binding protein. These genes were preferentially expressed in most of the cell lines of lung cancer and also of ovarian cancer and renal cell carcinoma at the mRNA level. The expression of these genes was confirmed in lung and other cancer tissue specimens. In normal tissues, clone 83 was expressed only in the colon, and hairpin-binding protein was not expressed at all, whereas BTBD2 was ubiquitously expressed. Clone 83, BTBD2, and hairpin-binding protein encoded two, one, and one epitope peptides that can be recognized by HLA-A2402-restricted CTLs, respectively. These epitope peptides possessed the ability to induce HLA A24-restricted tumor-specific CTLs after in vitro stimulation in a culture of peripheral blood mononuclear cells from patients with lung cancer. These results suggest that these genes and peptides are potential candidates for cancer vaccines in HLA-A24(+) patients with lung cancer. PMID- 12782589 TI - CD40-stimulated B lymphocytes pulsed with tumor antigens are effective antigen presenting cells that can generate specific T cells. AB - Although they are considered as antigen-presenting cells, the role of antigen unspecific B lymphocytes in antigen presentation and T-lymphocyte stimulation remains controversial. In this paper, we tested the capacity of normal human peripheral activated B cells to stimulate T cells using melanoma antigens or melanoma cell lysates. B lymphocytes activated through CD40 ligation and then pulsed with tumor antigens efficiently processed and presented MHC class II restricted peptides to specific CD4(+) T-cell clones. This suggests that CD40 activated B cells have the functional and molecular competence to present MHC class II epitopes when pulsed with exogenous antigens, thereby making them a relevant source of antigen-presenting cells to generate T cells. To test this hypothesis, CD40-activated B cells were pulsed with a lysate prepared from melanoma cells and used to stimulate peripheral autologous T cells. Interestingly, T cells specific to melanoma antigens were generated. Additional analysis of these T-cell clones revealed that they recognized MHC class II restricted epitopes from tyrosinase, a known melanoma tumor antigen. The efficient antigen presentation by antigen-unspecific activated B cells was correlated with a down-regulation in the expression of HLA-DO, a B cell-specific protein known to interfere with HLA-DM function. Because HLA-DM is important in MHC class II peptide loading, the observed decrease in HLA-DO may partially explain the enhanced antigen presentation after B-cell activation. Results globally suggest that when they are properly activated, antigen-unspecific B lymphocytes can present exogenous antigens by MHC class II molecules and stimulate peripheral antigen-specific T cells. Antigen presentation by activated B cells could be exploited for immunotherapy by allowing the in vitro generation of T cells specific against antigens expressed by tumors or viruses. PMID- 12782590 TI - Murine dendritic cells pulsed with an anti-idiotype antibody induce antigen specific protective antitumor immunity. AB - In this study, using the carcinoembryonic antigen (CEA)-expressing C15 murine colon carcinoma system in syngeneic C57BL/6 mice, we have evaluated the efficacy of bone marrow-derived dendritic cells (DCs) pulsed with the murine anti-idiotype antibody 3H1 as a tumor vaccine. Anti-idiotype 3H1 mimics a distinct and specific epitope of CEA and can generate anti-CEA immunity in mice, rabbits, monkeys, and humans when used with a conventional immune adjuvant. Our goal was to determine whether the use of DC as direct antigen-presenting cells would improve the potency of 3H1 as vaccine. Bone marrow-DC pulsed with 3H1 and injected into naive mice induced both humoral and cellular anti-3H1, as well as anti-CEA immunity. Specific killing of C15 cells in in vitro antibody-dependent cellular cytotoxicity has been observed by immune sera. Immune-splenic lymphocytes when stimulated in vitro with 3H1 or CEA, showed increased proliferative CD4(+) Th1 type T-cell response and secreted significantly high levels of Th1 cytokines [IFN gamma, interleukin (IL)-2] and low levels of Th2 cytokines (IL-4, IL-10). This vaccine also induced MHC class I antigen-restricted CD8(+) T-cell responses. The up-regulation of activation markers CD69 and CD25 on CD8(+) CTLs correlated with antigen-specific strong CTL responses in vitro. The immunity induced in mice resulted in a complete rejection of CEA-expressing C15 tumor cells in 100% of experimental mice, whereas no protection was observed when 3H1-pulsed DC vaccinated mice were challenged with CEA-negative MC-38 cells. The tumor rejection in 3H1-pulsed DC-treated mice was associated with the induction of a memory response that helped those mice to survive a second challenge with a lethal dose of C15 cells. PMID- 12782591 TI - Inhibition of BUB1 results in genomic instability and anchorage-independent growth of normal human fibroblasts. AB - The relative contribution of aneuploidy and gene mutations to human tumorigenesis is not yet known. Studies in mice have demonstrated that even single point mutations in oncogenes and tumor suppressor genes can dramatically increase tumor frequency. However, models to evaluate the definitive role of aneuploidy and genomic instability are not yet available. Human fibroblast cells have long been used as a tool for investigating proliferation, senescence, immortalization, and tumorigenesis, all processes that are strongly interrelated. We have now used antisense and ribozyme-mediated temporary inhibition of BUB1 to study the consequences of mitotic checkpoint failure on the development of aneuploidy. The analysis of cell colonies selected by soft agar growth showed evidence of chromosome instability and delayed senescence, without being tumorigenic in nude mice. Our data suggest that chromosomal instability and aneuploidy are early changes that precede tumorigenicity in the multistep process leading to neoplastic transformation. PMID- 12782592 TI - Caveolin-1 and caveolin-2,together with three bone morphogenetic protein-related genes, may encode novel tumor suppressors down-regulated in sporadic follicular thyroid carcinogenesis. AB - Thyroid cancer is common, occurring in 1% of the general population. The relative frequencies of two of the most common subtypes of thyroid carcinoma, follicular (FTC) and papillary (PTC), vary depending on the regional prevalence of iodine deficiency. Although PTC has been more extensively studied, the etiology of sporadic FTC is poorly understood. To further elucidate this, we conducted microarray expression comparison of FTC tumors and normal thyroid tissue. Three commonly down-regulated genes, caveolin-1, caveolin-2, and GDF10/BMP3b, were chosen for further study on the basis of their localization to two chromosomal regions, 7q31.1 and 10q11.1, that commonly show loss of heterozygosity in FTC. Two additional genes, glypican-3 and a novel chordin-like gene, were also analyzed in view of their involvement in bone morphogenetic protein signaling and possible interaction with GDF10. Each of these five genes was down-regulated in >or=15 of 19 FTC tumors (79%) by semiquantitative reverse transcription-PCR. Caveolin-1 showed preferential down-regulation of its beta-isoform at both the mRNA and protein level, suggesting a distinct function for this isoform. Caveolin 1 is of particular functional interest because it has been shown to interact with PTEN, the tumor suppressor gene mutated in Cowden syndrome, an inherited multiple hamartoma syndrome that includes predisposition to FTC. Immunohistochemical analysis of 141 thyroid tumors of various histological types showed significantly fewer caveolin-1-positive tumors in FTCs, including insular type tumors, and Hurthle cell carcinomas in comparison with normal thyroid. PTC and anaplastic thyroid carcinomas did not show significant down-regulation, and thus, caveolin-1 may become a useful molecular marker to differentiate the various histologies of thyroid malignancies. PMID- 12782593 TI - Array-based comparative genomic hybridization for genome-wide screening of DNA copy number in bladder tumors. AB - Genome-wide copy number profiles were characterized in 41 primary bladder tumors using array-based comparative genomic hybridization (array CGH). In addition to previously identified alterations in large chromosomal regions, alterations were identified in many small genomic regions, some with high-level amplifications or homozygous deletions. High-level amplifications were detected for 192 genomic clones, most frequently at 6p22.3 (E2F3), 8p12 (FGFR1), 8q22.2 (CMYC), 11q13 (CCND1, EMS1, INT2), and 19q13.1 (CCNE). Homozygous deletions were detected in 51 genomic clones, with four showing deletions in more than one case: two clones mapping to 9p21.3 (CDKN2A/p16, in nine cases), one at 8p23.1 (three cases), and one at 11p13 (two cases). Significant correlations were observed between copy number gain of clones containing CCNE1 and gain of ERBB2, and between gain of CCND1 and deletion of TP53. In addition, there was a significant complementary association between gain of CCND1 and gain of E2F3. Although there was no significant relationship between copy number changes and tumor stage or grade, the linked behavior among genomic loci suggests that array CGH will be increasingly important in understanding pathways critical to bladder tumor biology. PMID- 12782594 TI - Loss of PTEN promotes tumor development in malignant melanoma. AB - Loss of tumor suppressor genes on chromosome 10 plays an important role in the development of 30-60% of melanomas; however, the identity of these genes and the mechanisms by which loss of these genes leads to tumor formation remain uncertain. The phosphatase and tensin homologue deleted from chromosome 10 (PTEN) is one of the genes on chromosome 10 whose of which the loss or inactivation may play an important role in melanoma tumorigenesis, but functional studies directly demonstrating PTEN involvement in melanomas are necessary to confirm this role. To determine the biological importance of PTEN loss in melanomas, we established a novel model in which an intact chromosome 10 was transferred into melanoma cells lacking PTEN protein to express the protein at normal physiological levels and to measure the consequent effects on melanoma tumorigenesis. PTEN expression in these cells retarded tumor development in mice unless, by analogy with loss of heterozygosity, the PTEN gene was deleted or inactivated during tumor formation. Mechanistically, PTEN loss led to the activation of Akt, which consequently down regulated the apoptotic pathway of melanoma cells. In contrast, expression of PTEN attenuated Akt activation, thereby increasing sensitivity to apoptotic stimuli in cell culture and in vivo in animal models. This model demonstrated that PTEN loss is critical for melanoma tumorigenesis and allowed a dissection of the underlying mechanism by which PTEN loss facilitated melanoma tumor development. In summary, loss of PTEN reduces apoptosis and promotes cell survival, thereby favoring melanoma tumor formation. Thus, these observations provide an etiological basis for PTEN loss during the genesis of sporadic melanomas. PMID- 12782595 TI - Histone deacetylase inhibitors activate p21(WAF1) expression via ATM. AB - Histone deacetylase (HDAC) inhibitors are known to induce expression of genes such as p21(WAF1), thereby, leading to cell cycle arrest. In this work, we show that p21(WAF1) induction by HDAC inhibitors (depsipeptide and trichostatin A) is defective in Ataxia telangiectasia (AT) cells but normal in matched wild-type (WT) cells (human diploid fibroblasts). To verify the role of ATM in this effect, we show that ectopic expression of the WT ATM gene in an AT cell line fully restores p21(WAF1) induction by the HDAC inhibitors. Furthermore, because caffeine and wortmannin attenuate p21(WAF1) induction in WT cells, it is probable that the phosphatidylinositol 3'-kinase activity is essential for this process. Besides the p21(WAF1) promoter, activation of topoisomerase IIIalpha and SV40 promoters by the HDAC inhibitors are also decreased in the AT cell lines relative to WT cells; thus, these findings pertain to other promoters. Finally, despite the obvious induction deficiency of gene expression, the overall levels of H3 and H4 histone acetylation appear to be the same between AT and normal cells in response to HDAC inhibitor treatments. Taken together, the data indicate that ATM is involved in histone acetylation-mediated gene regulation. PMID- 12782596 TI - A novel single nucleotide polymorphism within the 5' tandem repeat polymorphism of the thymidylate synthase gene abolishes USF-1 binding and alters transcriptional activity. AB - Thymidylate synthase (TS) gene expression is modulated by a polymorphism in the 5' regulatory region of the gene. The polymorphism consists mainly of either two repeats (2R) or three repeats (3R) of a 28-bp sequence, yielding greater TS gene expression and protein levels with a 3R genotype. Two USF family E-box consensus elements are found within the tandem repeats of the 3R genotype, and one is found within the 2R genotype. These elements bind USF proteins in vitro by electrophoretic mobility shift analysis and in vivo by chromatin immunoprecipitation assay. We show that the additional USF consensus element within the 3R construct confers greater transcriptional activity relative to the 2R construct. Mutagenesis of the USF sites shows that the transcriptional regulation of TS is dependent, in part, on USF proteins binding within the tandem repeats. In addition, we identified a novel G-->C single nucleotide polymorphism in the second repeat of 3R alleles within the USF consensus element that alters the ability of USF proteins to bind and thus alters the transcriptional activation of TS gene constructs bearing this genotype. Through RFLP analysis, we determined the respective frequencies of the C allele (3RC) among all 3R alleles in non-Hispanic whites, Hispanic whites, African Americans, and Singapore Chinese to be 56%, 47%, 28%, and 37%, respectively. Based on our findings, this novel single nucleotide polymorphism should be considered when the 5' tandem repeat polymorphism is being used as a predictor of clinical outcome to TS inhibitors. PMID- 12782597 TI - Cooperation of two mutant p53 alleles contributes to Fas resistance of prostate carcinoma cells. AB - Both inactivation of p53 function and loss of sensitivity to Fas contribute to a malignant phenotype and frequently occur during tumor progression. Although in the majority of cases only one of the p53 alleles is mutated, some tumors acquire mutations in both alleles of the p53 gene. To determine the biological significance of this phenomenon, we analyzed p53 mutants, p53(223Leu) and p53(274Phe), from Fas-resistant prostate carcinoma cell line DU145. Both mutants differed from wild-type p53 in their conformation, transactivation ability, and effect on the growth of p53-deficient cells, with p53(223Leu) being more similar to wild-type p53 than was p53(274Phe). Interestingly, the biological effect of coexpression of the DU145-derived mutants was dramatically different from that of each mutant expressed alone. Whereas neither of the two mutants was found to be dominant-negative against wild-type p53, each neutralized the other's growth suppressive effects and, in combination, were capable of down-regulating Fas expression and converting Fas-sensitive prostate carcinoma cells PC3 into Fas resistant ones. These results indicate that two different p53 mutants that are separately rather weak can cooperate to generate p53 protein with anti-Fas function that is likely to provide additional selective advantages to the tumor. PMID- 12782598 TI - Novel candidate targets of beta-catenin/T-cell factor signaling identified by gene expression profiling of ovarian endometrioid adenocarcinomas. AB - The activity of beta-catenin (beta-cat), a key component of the Wnt signaling pathway, is deregulated in about 40% of ovarian endometrioid adenocarcinomas (OEAs), usually as a result of CTNNB1 gene mutations. The function of beta-cat in neoplastic transformation is dependent on T-cell factor (TCF) transcription factors, but specific genes activated by the interaction of beta-cat with TCFs in OEAs and other cancers with Wnt pathway defects are largely unclear. As a strategy to identify beta-cat/TCF transcriptional targets likely to contribute to OEA pathogenesis, we used oligonucleotide microarrays to compare gene expression in primary OEAs with mutational defects in beta-cat regulation (n = 11) to OEAs with intact regulation of beta-cat activity (n = 17). Both hierarchical clustering and principal component analysis based on global gene expression distinguished beta-cat-defective tumors from those with intact beta-cat regulation. We identified 81 potential beta-cat/TCF targets by selecting genes with at least 2-fold increased expression in beta-cat-defective versus beta-cat regulation-intact tumors and significance in a t test (P < 0.05). Seven of the 81 genes have been previously reported as Wnt/beta-cat pathway targets (i.e., BMP4, CCND1, CD44, FGF9, EPHB3, MMP7, and MSX2). Differential expression of several known and candidate target genes in the OEAs was confirmed. For the candidate target genes CST1 and EDN3, reporter and chromatin immunoprecipitation assays directly implicated beta-cat and TCF in their regulation. Analysis of presumptive regulatory elements in 67 of the 81 candidate genes for which complete genomic sequence data were available revealed an apparent difference in the location and abundance of consensus TCF-binding sites compared with the patterns seen in control genes. Our findings imply that analysis of gene expression profiling data from primary tumor samples annotated with detailed molecular information may be a powerful approach to identify key downstream targets of signaling pathways defective in cancer cells. PMID- 12782599 TI - The CCL6 chemokine is differentially regulated by c-Myc and L-Myc, and promotes tumorigenesis and metastasis. AB - The CCL6 chemokine gene was identified as a direct positive target of the L-Myc oncoprotein in interleukin 3-dependent 32D myeloid cells. A mutant form of c-Myc, lacking a region of the NH(2)-terminal domain necessary for transcriptional repression (c-MycDeltaMBII), also up-regulated CCL6. Chromatin immunoprecipitation showed that L-Myc, c-MycDeltaMBII, and full-length c-Myc all bound the CCL6 promoter, although the latter was inactive in transcriptional up regulation. Exogenously added CCL6 induced marked apoptosis in some cell types. However, in 32D cells, the coexpression of c-Myc and CCL6 abrogated interleukin 3 dependence and produced a highly leukemogenic phenotype. In two solid tumor models, CCL6 overexpression also accelerated tumor growth, and/or enhanced local and metastatic spread in association with marked apoptosis of the tumor capsule and adjacent normal tissues. Our results show that CCL6 can be either a positive or negative target for Myc oncoproteins. The chemokine may alter tumor behavior by relieving its growth factor dependency and by promoting invasiveness as a result of local tissue apoptosis. PMID- 12782600 TI - The 43000 growth-associated protein functions as a negative growth regulator in glioma. AB - Previous molecular analyses of human astrocytomas have identified many genetic changes associated with astrocytoma formation and progression. In an effort to identify novel gene expression changes associated with astrocytoma formation, which might reveal new potential targets for glioma therapeutic drug design, we used the B8-RAS-transgenic mouse astrocytoma model. Using multiplex gene expression profiling, we found that growth-associated protein 43 (GAP43) RNA and protein expression were lost in select human and mouse glioma cell lines. In this study, we demonstrate that re-expression of GAP43 in deficient C6 glioma cells results in growth suppression in clonogenic assays, as well as in multiple independently derived C6 glioma cell lines in vitro. GAP43-expressing C6 cells also exhibit reduced tumor growth as s.c. explants in immunocompromised mice in vivo. In addition, GAP43-expressing C6 clones demonstrate impaired cell motility and increased homophilic aggregation. GAP43 re-expression is also associated with reduced mitogen-activated protein kinase and AKT activation in C6 cells, suggesting that GAP43 functions as a novel glioma growth suppressor by modulating mitogenic signaling pathways. PMID- 12782601 TI - Blockade of epidermal growth factor receptor signaling leads to inhibition of renal cell carcinoma growth in the bone of nude mice. AB - Renal cell carcinoma (RCC) frequently produces metastases to the musculoskeletal system that are a major source of morbidity in the form of pain, immobilization, fractures, neurological compromise, and a decreased ability to perform activities of daily living. Patients with metastatic RCC therefore have a dismal prognosis because there is no effective adjuvant treatment for this disease. Because the epidermal growth factor receptor (EGF-R) signaling cascade is important in the growth and metastasis of RCC, its blockade has been hypothesized to inhibit tumor growth and hence prevent resultant bone destruction. We determined whether blockade of EGF-R by the tyrosine kinase inhibitor PKI 166 inhibited the growth of RCC in bone. We use a novel cell line, RBM1-IT4, established from a human RCC bone metastasis. Protein and mRNA expression of the ligands and receptors was assessed by Western and Northern blots. The stimulation of RBM1-IT4 cells with epidermal growth factor or transforming growth factor alpha resulted in increased cellular proliferation and tyrosine kinase autophosphorylation. PKI 166 prevented these effects. First, RBM1-IT4 cells were implanted into the tibia of nude mice, where they established lytic, progressively growing lesions, after which the mice were treated with PKI 166 alone or in combination with paclitaxel (Taxol). Immunohistochemical analysis revealed that tumor cells and tumor-associated endothelial cells in control mice expressed activated EGF-R. Treatment of mice with PKI 166 alone or in combination with Taxol produced a significant decrease in the incidence and size of bone lesions as compared with the results in control or Taxol-treated mice (P < 0.001). Treatment with PKI 166 also decreased the expression of phosphorylated EGF-R by tumor cells and tumor-associated endothelial cells, and this was even more pronounced with PKI 166 plus Taxol treatment. The PKI 166 plus Taxol combination produced apoptosis of tumor cells and tumor-associated endothelial cells. Tumor cell proliferation, shown by proliferating cell nuclear antigen positivity, was decreased in all treatment groups. In addition, the integrity of the bone was maintained in mice treated with PKI 166 or PKI 166 plus Taxol, whereas massive bone destruction was seen in control and Taxol-treated mice. These results suggest that blockade of EGF-R signaling inhibits growth of RCC in the bone by its effect on tumor cells and tumor-associated endothelial cells. PMID- 12782602 TI - Epidermal growth factor receptor-independent constitutive activation of STAT3 in head and neck squamous cell carcinoma is mediated by the autocrine/paracrine stimulation of the interleukin 6/gp130 cytokine system. AB - The aberrant growth of head and neck squamous cell carcinoma (HNSCC) is often associated with the constitutive activation of signal-transducer-and-activator-of transcription-3 (STAT3), which is believed to result from the persistent stimulation of EGF receptors that are highly expressed in squamous cell carcinoma (SCC) cells. To investigate the mechanism underlying STAT3 deregulation in HNSCC, we examined the interplay of the STAT3 and epidermal growth factor receptor (EGFR) signaling pathways using a panel of HNSCC cell lines. Although STAT3 was active in most HNSCC cell lines, only 3 of 10 HNSCC cell lines were moderately to strongly positive for activated EGFR. Even in the EGFR-positive cell lines, STAT3 activation was not dependent on EGFR activation, as STAT3 tyrosine phosphorylation levels persisted after treatment with AG1478, a chemical inhibitor of EGFR activity. Furthermore, we found that conditioned medium harvested from HNSCC cells could induce STAT3 tyrosine phosphorylation in immortalized keratinocytes regardless of the status of EGFR signaling. In contrast, blocking the cytokine gp130 coreceptor abolished STAT3 tyrosine phosphorylation in HNSCC cells and that induced by the conditioned medium. Immunodepletion studies suggested interleukin 6 (IL6) as the major autocrine/paracrine factor for STAT3 activation, which coincided with high levels of secretion of IL6 into the culture medium by these cancer cells. Treatment with a specific inhibitor of Janus kinase, AG490, in HNSCC cells led to a reduction of active STAT3 and caused significant growth retardation and apoptosis. Thus, constitutive activation of STAT3 in HNSCC may use an autocrine/paracrine activating loop mediated by IL6 and other cytokines acting through the gp130 receptor family, which may confer both proliferative and survival potential in this malignancy. PMID- 12782603 TI - Overexpression of legumain in tumors is significant for invasion/metastasis and a candidate enzymatic target for prodrug therapy. AB - Expression of legumain, a novel asparaginyl endopeptidase, in tumors was identified from gene expression profiling and tumor tissue array analysis. Legumain was demonstrated in membrane-associated vesicles concentrated at the invadopodia of tumor cells and on cell surfaces where it colocalized with integrins. Legumain was demonstrated to activate progelatinase A. Cells overexpressing legumain possessed increased migratory and invasive activity in vitro and adopted an invasive and metastatic phenotype in vivo, inferring significance of legumain in tumor invasion and metastasis. A prodrug strategy incorporating a legumain-cleavable peptide substrate onto doxorubicin was developed. The prototype compound, designated legubicin, exhibited reduced toxicity and was effectively tumoricidal in vivo in a murine colon carcinoma model. PMID- 12782604 TI - Localization and characterization of melanoma-associated glycosaminoglycans: differential expression of chondroitin and heparan sulfate epitopes in melanoma. AB - Glycosaminoglycans (GAGs) are anionic polysaccharides present on cells and in the extracellular matrix (ECM). They likely play a role in tumor formation because of their capacity to bind and modulate a variety of proteins including growth factors, cytokines, and proteases. Using a panel of (human) phage display-derived anti-GAG antibodies, the location and expression of GAG epitopes in human cutaneous melanocytic lesions was studied. Antibodies EW4E1 and EW4G2 identified a melanoma-associated chondroitin sulfate/heparan sulfate epitope, whereas antibody EW4B7 recognized a melanoma-associated heparan sulfate epitope. These antibodies showed a high reactivity with blood vessels and ECM in cutaneous melanoma tumors, whereas their reactivity with nevi was very low. Using a set of defined oligosaccharides it was established that sulfate groups are of main importance in the binding to the antibodies and that glycomimetics can mimic natural oligosaccharides. In xenografts of melanoma cell line MeL57, a strong association of GAG epitopes with an injected fluorescent fluid flow tracer was observed. In uveal melanoma antibody, EW4E1 proved to be a sensitive probe for the detection of the geometry of ECM structures, known to have prognostic value. Taken together, data indicate that in melanoma a defined set and location of GAG epitopes are present with possible functional significance. PMID- 12782605 TI - Tissue-specific microvascular endothelial cell lines from H-2K(b)-tsA58 mice for studies of angiogenesis and metastasis. AB - Microvascular endothelial cells play a critical role in tumor progression and metastasis by forming capillary networks that encourage tumor growth and by promoting the attachment of circulating tumor cells to the vascular wall of distant tissues. Efforts to study the molecular mechanisms that mediate these complex processes in different anatomical compartments have been impeded by difficulties in the isolation and propagation of endothelial cells from different organs. To overcome these limitations, we used two-color flow cytometry to identify and select microvascular endothelial cells from primary cultures obtained from different organs of mice whose tissues harbor a temperature sensitive SV40 large T antigen (H-2K(b)-tsA58 mice; ImmortoMice). The selection strategy targeted cell populations expressing the inducible endothelial cell adhesion molecules, E-selectin and VCAM-1, and proved successful in generating microvascular endothelial cell lines from a number of different organs. When cultured under permissive temperatures (33 degrees C), individual cell lines displayed doubling times consistent with endothelial cells possessing an angiogenic phenotype. The transfer of endothelial cells to nonpermissive temperatures (37 degrees C) resulted in cell differentiation and the induction of a quiescent state. Established cell lines exhibited several inherent endothelial properties, including the expression of constitutive and inducible levels of endothelial cell adhesion molecules E-selectin, intercellular adhesion molecule 1, and vascular cell adhesion molecule-1, internalization of acetylated low density lipoprotein, and formation of loop structures on Matrigel surfaces. The immortalized endothelial cell lines established from H-2K(b)-tsA58 mice provide, for the first time, a cell culture system to examine factors regulating angiogenesis and tumor cell arrest in different organ systems. PMID- 12782606 TI - Jun activation domain-binding protein 1 expression in breast cancer inversely correlates with the cell cycle inhibitor p27(Kip1). AB - The Jun activation domain-binding protein 1 (JAB1), aside from being an activator protein 1 coactivator, is involved in degradation of the cyclin-dependent kinase inhibitor p27. We examined JAB1 and p27 protein expression in invasive breast carcinoma specimens and the association of this expression with clinical outcome. JAB1 was detected immunohistochemically in 43 of 53 (81%) tumors; 32 (60%) breast carcinomas showed high JAB1 expression (>50% of cells positive) and reduced or absent p27 levels (P = 0.02, Mann-Whitney U test). Tumors with high p27 expression were rarely positive for JAB1. All eight patients with JAB1-negative tumors had no evidence of relapse or disease progression at a median follow-up of 70 months. Immunoblotting showed strong JAB1 expression in breast carcinoma samples but not in paired normal breast epithelial samples. Targeted overexpression of JAB1 by regulated adenovirus in breast cancer cell lines also reduced p27 levels by accelerating degradation of p27. Thus, the JAB1:p27 ratio may be a novel indicator of aggressive, high-grade tumor behavior, and control of JAB1 could be a novel target for experimental therapies. PMID- 12782607 TI - Latent membrane protein 1 is critical for efficient growth transformation of human B cells by epstein-barr virus. AB - The EBV latent membrane protein 1 (LMP1) is an integral membrane protein that acts like a constitutively activated receptor. LMP1 interacts with members of the tumor necrosis factor receptor-associated factor family, as well as with tumor necrosis factor receptor-associated death domain, resulting in induction of nuclear factor-kappaB, the p38 mitogen-activated protein kinase pathway, and the c-Jun NH(2)-terminal kinase activator protein 1-signaling cascade. The binding of Janus kinase 3 results in activation of signal transducers and activators of transcription. The domain structure of LMP1 has been mapped extensively, but the quantitative contribution of distinct LMP1 domains to the efficiency of B-cell proliferation by EBV has not been determined. On the basis of the maxi-EBV system, which allows us to introduce and study mutations in the context of the complete EBV genome, a panel of 10 EBV mutants with alterations in the LMP1 gene locus was established. The mutant EBVs were tested for their efficiency to induce and maintain proliferation of clonal B-cell lines in vitro. Surprisingly and with reduced frequency, EBV mutants which deleted LMP1's COOH terminus, transmembrane domains, or the entire open reading frame were able to generate proliferating B cell clones that were dependent on the presence of human fibroblast feeder cells. A B-cell clone carrying the LMP1-null mutant EBV genome was also analyzed for oncogenicity in severe combined immunodeficiency mice. Our results demonstrate that LMP1 is critical but not mandatory for the generation of proliferating B cells in vitro. LMP1 functions greatly contribute to EBV's transformation potential and appear essential for its oncogenicity in severe combined immunodeficiency mice. PMID- 12782608 TI - Down-regulation of c-Met inhibits growth in the liver of human colorectal carcinoma cells. AB - Overexpression of c-Met, the protein tyrosine kinase receptor for the hepatocyte growth factor/scatter factor, has been implicated in the progression and metastasis of human colorectal carcinoma. To examine the role of c-Met on in vitro and in vivo growth of human colon tumor cell lines, stable subclones of the high metastatic human colorectal carcinoma cell line, KM20, isolated from a Dukes' D patient, with reduced c-Met expression were obtained after transfection with a c-Met-specific targeting ribozyme. The subclones were only modestly reduced in c-Met expression because of c-Met playing an important role in cellular survival. However, a 60-90% reduction in basal c-Met autophosphorylation and kinase activity were observed. Correlating with the reduction in c-Met kinase activity, subclones with reduced c-Met expression had significantly reduced in vitro growth rates and soft-agar colony-forming abilities. The in vivo growth of these cells was examined at both the ectopic SQ site and the orthotopic site of metastatic growth, the liver. SQ growth was delayed significantly in the c-Met down-regulated clones compared with controls, with tumors growing on loss of the ribozyme construct. In contrast, tumor incidence was significantly reduced when the c-Met down-regulated cells were grown in the orthotopic liver site. Thus, c Met activation may be important in metastatic growth of colon tumor cells in the liver. Collectively these data demonstrate that a small reduction in c-Met protein levels leads to profound biological effects, and potential c-Met inhibitors may be of therapeutic value in treatment of colon cancer. PMID- 12782609 TI - Tissue factor/factor VIIa inhibitors block angiogenesis and tumor growth through a nonhemostatic mechanism. AB - An association between cancer and thrombosis has been recognized for more than a century. However, the manner by which tumor growth is regulated by coagulation in vivo remains unclear. To assess the role of coagulation on tumor growth, in vivo, we tested coagulation inhibitors specific for either tissue factor (TF)/factor VIIa (fVIIa) complexes or factor Xa (fXa) for antitumor activity. Here, we show that two inhibitors of TF/fVIIa, TF pathway inhibitor (TFPI) and the nematode anticoagulant protein rNAPc2, inhibit both primary and metastatic tumor growth in mice. In addition, we show that rNAPc2 is also a potent inhibitor of angiogenesis. In contrast, rNAP5, a second nematode anticoagulant protein that specifically inhibits fXa, does not exhibit antitumor activity. Because the hemostatic activity of TF/fVIIa is mediated through activation of fXa, these data suggest that proteolytic activity of TF/fVIIa promotes tumor growth and angiogenesis through a novel proangiogenic mechanism and independently of hemostasis. PMID- 12782610 TI - Mouse models of human cancer consortium symposium on nervous system tumors. AB - Nervous system tumors represent unique neoplasms that arise within the central and peripheral nervous system. Recent progress in generating genetically engineered mouse models of these tumors has advanced our understanding of the critical molecular and cellular events important for the development of these tumors. Recently, the National Cancer Institute-sponsored Mouse Models of Human Cancer Consortium convened a meeting on Nervous System Tumors to review recent advances and suggest directions for future research. Refined and novel approaches to modeling central nervous system tumors, including gliomas, meningiomas, medulloblastomas, and oligodendrogliomas, as well as peripheral nervous system tumors such as neurofibromas, schwannomas, and malignant peripheral nerve sheath tumors, were presented. In this review, we discuss the current status of mouse modeling of human nervous system cancers with a specific focus on unresolved scientific questions pertaining to the molecular genetics and cellular biology of these tumors. PMID- 12782611 TI - Fourteenth Annual Pezcoller Symposium: the novel dichotomy of immune interactions with tumors. AB - The main focus of the Symposium was the fact that cell types of the innate and adaptive immune systems can have tumor-favoring as well as tumor antagonistic effects, both in a preventive and therapeutic mode. It was shown that macrophages (Mphi) and dendritic cells within a tumor exert tumor-favoring effects through the action of certain cytokines. Inflammatory reactions could favor the onset and growth of tumors. Dual immune functions were shown with CD4+ T cells and certain matrix metalloproteinase (MMP) activities favoring tumor progression and CD8+ T cells and certain heat shock proteins having antitumor action. Lack of antitumor action despite positive immune stimulation was also shown to depend on the existence of barriers to tumor infiltration by lymphocytes; remodeling of vasculature, e.g., by IFNgamma-induced cytokines like MIG and IPIO, reversed this type of impediment. Certain CXC cytokines increased tumor progression, whereas others, particularly those induced by IFNgamma, had the opposite effect; stromal derived factor-1 and its receptor CXCR4 affected tumor propensity to metastasize in certain organs. Stromal-derived factor-1 induced MMP9, which in turn regulated the bioavailability of vascular endothelial growth factor and the cascade of its tumor-favoring effects, whereas granulocyte colony-stimulating factor decreased MMP9 and the consequences of its action. The effects of certain proinflammatory cytokines and vascular endothelial growth factor functions in angiogenesis and lymphoangiogenesis were also discussed. The favoring effects of fever-like thermal stress on the function of molecules instrumental in lymphoid cell adhesion to vessels and infiltration into sites of immune actions were described. The mechanisms involved in the development of immune memory and those conditioning Type I and CTL responses were also discussed. A number of presentations were concerned with laboratory studies aimed at developing clinical regimens with potential activity in the prevention or treatment of cancer. Prevention of Her2/neu breast cancer in transgenic mice was achieved by suitable regimens with IL12 combined with vaccines, including DNA-based vaccines administered in conjunction with electroporation. Vaccination with shared tumor antigen MUCI or cyclin B was discussed, and its clinical translation was described. The prevention of TRAMP prostate tumor in transgenic mice by anti CTLA4 antibody plus vaccine was described, as was the translation of these regimens to the clinics. Clinical successes in melanoma patients using antimelanoma antigen antibodies in a therapeutic mode and precautions to be exerted in evaluating in vivo immune responses based on in vitro assays were emphasized. The symposium was concluded with an overall discussion focused on basic questions related to the capability of immunity to exert tumor-favoring or antitumor effects depending on conditions determined by both tumor and host functions. PMID- 12782612 TI - The fish story: a diet-heart hypothesis with clinical implications: n-3 polyunsaturated fatty acids, myocardial vulnerability, and sudden death. PMID- 12782613 TI - Endothelial recovery: the next target in restenosis prevention. PMID- 12782614 TI - The way to a human's heart is through the stomach: visceral endoderm-like cells drive human embryonic stem cells to a cardiac fate. PMID- 12782615 TI - Unstable angina and non-ST-elevation myocardial infarction: initial antithrombotic therapy and early invasive strategy. PMID- 12782616 TI - Clinical prevention of sudden cardiac death by n-3 polyunsaturated fatty acids and mechanism of prevention of arrhythmias by n-3 fish oils. PMID- 12782617 TI - Images in cardiovascular medicine. Electroanatomic mapping of cardiac resynchronization therapy. PMID- 12782618 TI - Images in cardiovascular medicine. Left ventricular asynchrony caused by an intramuscular lipoma: computed tomographic and magnetic resonance detection. PMID- 12782620 TI - The R-SNARE motif of tomosyn forms SNARE core complexes with syntaxin 1 and SNAP 25 and down-regulates exocytosis. AB - Tomosyn is a 130-kDa syntaxin-binding protein that contains a large N-terminal domain with WD40 repeats and a C-terminal domain homologous to R-SNAREs. Here we show that tomosyn forms genuine SNARE core complexes with the SNAREs syntaxin 1 and SNAP-25. In vitro studies with recombinant proteins revealed that complex formation proceeds from unstructured monomers to a stable four-helical bundle. The assembled complex displayed features typical for SNARE core complexes, including a profound hysteresis upon unfolding-refolding transitions. No stable complexes were formed between the SNARE motif of tomosyn and either syntaxin or SNAP-25 alone. Furthermore, both native tomosyn and its isolated C-terminal domain competed with synaptobrevin for binding to endogenous syntaxin and SNAP-25 on inside-out sheets of plasma membranes. Tomosyn-SNARE complexes were effectively disassembled by the ATPase N-ethylmaleimide-sensitive factor together with its cofactor alpha-SNAP. Moreover, the C-terminal domain of tomosyn was as effective as the cytoplasmic portion of synaptobrevin in inhibiting evoked exocytosis in a cell-free preparation derived from PC12 cells. Similarly, overexpression of tomosyn in PC12 cells resulted in a massive reduction of exocytosis, but the release parameters of individual exocytotic events remained unchanged. We conclude that tomosyn is a soluble SNARE that directly competes with synaptobrevin in the formation of SNARE complexes and thus may function in down-regulating exocytosis. PMID- 12782621 TI - Phosphatidylinositol phosphate kinase type 1gamma and beta1-integrin cytoplasmic domain bind to the same region in the talin FERM domain. AB - Talin is an essential component of focal adhesions that couples beta-integrin cytodomains to F-actin and provides a scaffold for signaling proteins. Recently, the integrin beta3 cytodomain and phosphatidylinositol phosphate (PIP) kinase type 1gamma (a phosphatidylinositol 4,5-bisphosphate-synthesizing enzyme) were shown to bind to the talin FERM domain (subdomain F3). We have characterized the PIP kinase-binding site by NMR using a 15N-labeled talin F2F3 polypeptide. A PIP kinase peptide containing the minimal talin-binding site formed a 1:1 complex with F2F3, causing a substantial number of chemical shift changes. In particular, two of the three Arg residues (Arg339 and Arg358), four of eight Ile residues, and one of seven Val residues in F3 were affected. Although a R339A mutation did not affect the exchange kinetics, R358A or R358K mutations markedly weakened binding. The Kd for the interaction determined by Trp fluorescence was 6 microm, and the R358A mutation increased the Kd to 35 microm. Comparison of these results with those of the crystal structure of a beta3-integrin cytodomain talin F2F3 chimera shows that both PIP kinase and integrins bind to the same surface of the talin F3 subdomain. Indeed, binding of talin present in rat brain extracts to a glutathione S-transferase integrin beta1-cytodomain polypeptide was inhibited by the PIP kinase peptide. The results suggest that ternary complex formation with a single talin FERM domain is unlikely, although both integrins and PIP kinase may bind simultaneously to the talin anti-parallel dimer. PMID- 12782622 TI - An endogenous inhibitor of focal adhesion kinase blocks Rac1/JNK but not Ras/ERK dependent signaling in vascular smooth muscle cells. AB - Humoral factors and extracellular matrix are critical co-regulators of smooth muscle cell (SMC) migration and proliferation. We reported previously that focal adhesion kinase (FAK)-related non-kinase (FRNK) is expressed selectively in SMC and can inhibit platelet-derived growth factor BB homodimer (PDGF-BB)-induced proliferation and migration of SMC by attenuating FAK activity. The goal of the current studies was to identify the mechanism by which FAK/FRNK regulates SMC growth and migration in response to diverse mitogenic signals. Transient overexpression of FRNK in SMC attenuated autophosphorylation of FAK at Tyr-397, reduced Src family-dependent tyrosine phosphorylation of FAK at Tyr-576, Tyr-577, and Tyr-881, and reduced phosphorylation of the FAK/Src substrates Cas and paxillin. However, FRNK expression did not alter the magnitude or dynamics of ERK activation induced by PDGF-BB or angiotensin II. Instead, FRNK expression markedly attenuated PDGF-BB-, angiotensin II-, and integrin-stimulated Rac1 activity and attenuates downstream signaling to JNK. Importantly, constitutively active Rac1 rescued the proliferation defects in FRNK expressing cells. Based on these observations, we hypothesize that FAK activation is required to integrate integrin signals with those from receptor tyrosine kinases and G protein-coupled receptors through downstream activation of Rac1 and that in SMC, FRNK may control proliferation and migration by buffering FAK-dependent Rac1 activation. PMID- 12782623 TI - Functional insights revealed by the crystal structures of Escherichia coli glucose-1-phosphatase. AB - The Escherichia coli periplasmic glucose-1-phosphatase is a member of the histidine acid phosphatase family and acts primarily as a glucose scavenger. Previous substrate profiling studies have demonstrated some of the intriguing properties of the enzyme, including its unique and highly selective inositol phosphatase activity. The enzyme is also potentially involved in pathogenic inositol phosphate signal transduction pathways via type III secretion into the host cell. We have determined the crystal structure of E. coli glucose-1 phosphatase in an effort to unveil the structural mechanism underlying such unique substrate specificity. The structure was determined by the method of multiwavelength anomalous dispersion using a tungstate derivative together with the H18A inactive mutant complex structure with glucose 1-phosphate at 2.4-A resolution. In the active site of glucose-1-phosphatase, there are two unique gating residues, Glu-196 and Leu-24, in addition to the conserved features of histidine acid phosphatases. Together they create steric and electrostatic constraints responsible for the unique selectivity of the enzyme toward phytate and glucose-1-phosphate as well as its unusually high pH optimum for the latter. Based on the structural characterization, we were able to derive simple structural principles that not only precisely explains the substrate specificity of glucose-1-phosphatase and the hydrolysis products of various inositol phosphate substrates but also rationalizes similar general characteristics across the histidine acid phosphatase family. PMID- 12782624 TI - Heparan sulfate 6-o-sulfotransferase is essential for muscle development in zebrafish. AB - Heparan sulfate proteoglycans function in development and disease. They consist of a core protein with attached heparan sulfate chains that are altered by a series of carbohydrate-modifying enzymes and sulfotransferases. Here, we report on the identification and characterization of a gene encoding zebrafish heparan sulfate 6-O-sulfotransferase (hs6st) that shows high homology to other heparan sulfate 6-O-sulfotransferases. When expressed as a fusion protein in cultured cells, the protein shows specific 6-O-sulfotransferase activity and preferentially acts on the iduronosyl N-sulfoglycosamine. In the developing embryo, hs6st is expressed in the brain, the somites, and the fins; the same structures that were affected upon morpholino-mediated functional knockdown. Morpholino injections significantly inhibited 6-O- but not 2-O-sulfation as assessed by HPLC. Morphants display disturbed somite specification independent of the somite oscillator mechanism and have impaired muscle differentiation. In conclusion, our results show that transfer of sulfate to specific positions on glycosaminoglycans is essential for muscle development. PMID- 12782625 TI - Sox8 is a specific marker for muscle satellite cells and inhibits myogenesis. AB - Sox8 belongs to a family of transcription regulators characterized by a unique DNA-binding domain known as the high mobility group box. Many Sox proteins play fundamental roles in vertebrate development and differentiation processes. Expression of Sox8 is strong during embryonic muscle development and gradually declines postnatally. In this study, we report that in adult skeletal muscle Sox8 is confined to satellite cells. Down-regulation during myogenic differentiation was also detected in cell culture systems and occurred in parallel with down regulation of the related Sox9. Overexpression of Sox8 or Sox9 on the other hand disrupted myoblasts in their ability to form myotubes. Concomitantly, expression of MyoD and myogenin decreased and basal as well as MyoD-induced activities of the myogenin promoter were strongly reduced in a Sox8-dependent manner. Our data suggest that Sox8 acts as a specific negative regulator of skeletal muscle differentiation, possibly by interfering with the function of myogenic basic helix-loop-helix proteins. PMID- 12782626 TI - Enzymes encoded by the farnesyl diphosphate synthase gene family in the Big Sagebrush Artemisia tridentata ssp. spiciformis. AB - Farnesyl diphosphate synthase catalyzes the sequential head-to-tail condensation of two molecules of isopentenyl diphosphate with dimethylallyl diphosphate. In plants the presence of farnesyl diphosphate synthase isozymes offers the possibility of differential regulation. Three full-length cDNAs encoding putative isoprenoid synthases, FDS-1, FDS-2, and FDS-5, with greater than 89% similarity were isolated from a Big Sagebrush Artemisia tridentata cDNA library using a three-step polymerase chain reaction protocol. One of the open reading frames, FDS-5, encoded a protein with an N-terminal amino acid extension that was identified as a plastidial targeting peptide. Recombinant histidine-tagged versions of three proteins were purified, and their enzymatic properties were characterized. FDS-1 and FDS-2 synthesized farnesyl diphosphate as the final chain elongation product, but their kinetic behavior varied. FDS-1 prefers geranyl diphosphate over dimethylallyl diphosphate as an allylic substrate and is active at acidic pH values compared with FDS-2. In contrast, FDS-5 synthesized two irregular monoterpenoids, chrysanthemyl diphosphate and lavandulyl diphosphate, when incubated with dimethylallyl diphosphate and an additional product, the regular monoterpene geranyl diphosphate, when incubated with isopentenyl diphosphate and dimethylallyl diphosphate. Specific cellular functions are proposed for each of the three enzymes, and a scenario for evolution of isoprenyl synthases in plants is presented. PMID- 12782628 TI - Protein kinase C epsilon dependence of the recovery from down-regulation of S1P1 G protein-coupled receptors of T lymphocytes. AB - Sphingosine 1-phosphate (S1P) from mononuclear phagocytes and platelets signals T cells predominantly through S1P1 G protein-coupled receptors (Rs) to enhance survival, stimulate and suppress migration, and inhibit other immunologically relevant responses. Cellular S1P1 Rs and their signaling functions are rapidly down-regulated by S1P, through a protein kinase C (PKC)-independent mechanism, but characteristics of cell-surface re-expression of down-regulated S1P1 Rs have not been elucidated. T cell chemotactic responses (CT) to 10 and 100 nm S1P and inhibition of T cell chemotaxis to chemokines (CI) by 1 and 3 microm S1P were suppressed after 1 h of preincubation with 100 nm S1P, but recovered fully after 12-24 h of exposure to S1P. Late recovery of down-regulated CT and CI, but not early down-regulation, was suppressed by PKC and PKCepsilon-selective inhibitors and was absent in T cells from PKCepsilon-null mice. The same PKCepsilon inhibitors blocked S1P-evoked increases in T cell nuclear levels of c-Fos and phosphorylated c-Jun and JunD after 24 h, but not 1 h. A mixture of c-Fos plus c Jun antisense oligonucleotides prevented late recovery of down-regulated CT and CI, without affecting S1P induction of down-regulation. Similarly, S1P-elicited threonine phosphorylation of S1P1 Rs was suppressed by a selective inhibitor of PKCepsilon after 24 h, but not 1 h. Biochemical requisites for recovery of down regulated S1P1 Rs thus differ from those for S1P induction of down-regulation. PMID- 12782627 TI - Potentiation of tumor necrosis factor alpha-induced secreted phospholipase A2 (sPLA2)-IIA expression in mesangial cells by an autocrine loop involving sPLA2 and peroxisome proliferator-activated receptor alpha activation. AB - In rat mesangial cells, exogenously added secreted phospholipases A2 (sPLA2s) potentiate the expression of pro-inflammatory sPLA2-IIA first induced by cytokines like tumor necrosis factor-alpha (TNFalpha) and interleukin-1 beta. The transcriptional pathway mediating this effect is, however, unknown. Because products of PLA2 activity are endogenous activators of peroxisome proliferator activated receptor alpha (PPAR alpha, we postulated that sPLA2s mediate their effects on sPLA2-IIA expression via sPLA2 activity and subsequent PPAR alpha activation. This study shows that various sPLA2s, including venom enzymes, human sPLA2-IIA, and wild-type and catalytically inactive H48Q mutant of porcine pancreatic sPLA2-IB, enhance the TNF alpha-induced sPLA2-IIA expression at the mRNA and protein levels. In cells transfected with luciferase sPLA2-IIA promoter constructs, sPLA2s are active only when the promoter contains a functional PPRE-1 site. The effect of exogenous sPLA2s is also blocked by the PPAR alpha inhibitor MK886. Interestingly, the expression of sPLA2-IIA induced by TNF alpha alone is also attenuated by MK886, by the sPLA2-IIA inhibitor LY311727, by heparinase, which prevents the binding of sPLA2-IIA to heparan sulfate proteoglycans, and by the specific cPLA2-alpha inhibitor pyrrolidine-1. Together, these data indicate that sPLA2-IIA released from mesangial cells by TNF alpha stimulates its own expression via an autocrine loop involving cPLA2 and PPAR alpha. This signaling pathway is also used by exogenously added sPLA2s including pancreatic sPLA2-IB and is distinct from that used by TNF alpha. PMID- 12782629 TI - The Gla domain of factor IXa binds to factor VIIIa in the tenase complex. AB - During blood coagulation factor IXa binds to factor VIIIa on phospholipid membranes to form an enzymatic complex, the tenase complex. To test whether there is a protein-protein contact site between the gamma-carboxyglutamic acid (Gla) domain of factor IXa and factor VIIIa, we demonstrated that an antibody to the Gla domain of factor IXa inhibited factor VIIIa-dependent factor IXa activity, suggesting an interaction of the factor IXa Gla domain with factor VIIIa. To study this interaction, we synthesized three analogs of the factor IXa Gla domain (FIX1-47) with Phe-9, Phe-25, or Val-46 replaced, respectively, with benzoylphenylalanine (BPA), a photoactivatable cross-linking reagent. These factor IX Gla domain analogs maintain native tertiary structure, as demonstrated by calcium-induced fluorescence quenching and phospholipid binding studies. In the absence of phospholipid membranes, FIX1-47 was able to inhibit factor IXa activity. This inhibition is dependent on the presence of factor VIIIa, suggesting a contact site between the factor IXa Gla domain and factor VIIIa. To demonstrate a direct interaction we did cross-linking experiments with FIX1 479BPA, FIX1-4725BPA, and FIX1-4746BPA. Covalent cross-linking to factor VIIIa was observed primarily with FIX1-4725BPA and to a much lesser degree with FIX1 4746BPA. Immunoprecipitation experiments with an antibody to the C2 domain of factor VIIIa indicate that the factor IX Gla domain cross-links to the A3-C1-C2 domain of factor VIIIa. These results suggest that the factor IXa Gla domain contacts factor VIIIa in the tenase complex through a contact site that includes phenylalanine 25 and perhaps valine 46. PMID- 12782630 TI - Exchange factors of the RasGRP family mediate Ras activation in the Golgi. AB - H-Ras and N-Ras become activated both at the plasma membrane and in endomembrane structures such as the Golgi apparatus. This compartmentalized activation is relevant from a signaling standpoint, because effector molecules can become activated differently depending on the region of the cell where Ras proteins are activated. An unsolved question in this new regulatory mechanism is the understanding of how Ras proteins become activated in endomembranes. To approach this problem, we have studied the subcellular distribution and activities of a number of Ras guanosine nucleotide exchange factors. Our results indicate that Ras activation at the plasma membrane and endoplasmic reticulum is an unspecific process that can be achieved by most Ras activators. In contrast, GTP loading of Ras at the Golgi is only induced by members of the Ras guanosine nucleotide releasing protein family. In agreement with these observations, Ras guanosine nucleotide releasing proteins are the only Ras activators showing localization in the Golgi. These results indicate that the compartmentalized activation of effector pathways by Ras proteins depends not only on the specific localization of the GTPases but also in the availability of GDP/GTP exchange factors capable of activating Ras proteins in specific subcellular compartments. PMID- 12782631 TI - Structure of the yeast cytochrome bc1 complex with a hydroxyquinone anion Qo site inhibitor bound. AB - Bifurcated electron transfer during ubiquinol oxidation is the key reaction of cytochrome bc1 complex catalysis. Binding of the competitive inhibitor 5-n-heptyl 6-hydroxy-4,7-dioxobenzothiazole to the Qo site of the cytochrome bc1 complex from Saccharomyces cerevisiae was analyzed by x-ray crystallography. This alkylhydroxydioxobenzothiazole is bound in its ionized form as evident from the crystal structure and confirmed by spectroscopic analysis, consistent with a measured pKa = 6.1 of the hydroxy group in detergent micelles. Stabilizing forces for the hydroxyquinone anion inhibitor include a polarized hydrogen bond to the iron-sulfur cluster ligand His181 and on-edge interactions via weak hydrogen bonds with cytochrome b residue Tyr279. The hydroxy group of the latter contributes to stabilization of the Rieske protein in the b-position by donating a hydrogen bond. The reported pH dependence of inhibition with lower efficacy at alkaline pH is attributed to the protonation state of His181 with a pKa of 7.5. Glu272, a proposed primary ligand and proton acceptor of ubiquinol, is not bound to the carbonyl group of the hydroxydioxobenzothiazole ring but is rotated out of the binding pocket toward the heme bL propionate A, to which it is hydrogen bonded via a single water molecule. The observed hydrogen bonding pattern provides experimental evidence for the previously proposed proton exit pathway involving the heme propionate and a chain of water molecules. Binding of the alkyl-6-hydroxy-4,7-dioxobenzothiazole is discussed as resembling an intermediate step of ubiquinol oxidation, supporting a single occupancy model at the Qo site. PMID- 12782632 TI - Cathepsin D triggers Bax activation, resulting in selective apoptosis-inducing factor (AIF) relocation in T lymphocytes entering the early commitment phase to apoptosis. AB - Activated human T lymphocytes exposed to apoptotic stimuli targeting mitochondria (i.e. staurosporine), enter an early, caspase-independent phase of commitment to apoptosis characterized by cell shrinkage and peripheral chromatin condensation. We show that during this phase, AIF is selectively released from the intermembrane space of mitochondria, and that Bax undergo conformational change, relocation to mitochondria, and insertion into the outer mitochondrial membrane, in a Bid-independent manner. We analyzed the subcellular distribution of cathepsins (Cat) B, D, and L, in a search for caspase-independent factors responsible for Bax activation and AIF release. All were translocated from lysosomes to the cytosol, in correlation with limited destabilization of the lysosomes and release of lysosomal molecules in a size selective manner. However, only inhibition of Cat D activity by pepstatin A inhibited the early apoptotic events and delayed cell death, even in the presence of bafilomycin A1, an inhibitor of vacuolar type H+-ATPase, which inhibits acidification in lysosomes. Small interfering RNA-mediated gene silencing was used to inactivate Cat D, Bax, and AIF gene expression. This allowed us to define a novel sequence of events in which Cat D triggers Bax activation, Bax induces the selective release of mitochondrial AIF, and the latter is responsible for the early apoptotic phenotype. PMID- 12782634 TI - Insulin recruits GLUT4 from distinct compartments via distinct traffic pathways with differential microtubule dependence in rat adipocytes. AB - In the present study, we investigated the physiological significance of the microtubules in the subcellular localization and trafficking of GLUT4 in rat primary adipocytes. Morphological and biochemical analyses revealed a dose- and time-dependent disruption of the microtubules by treatment with nocodazole. With nearly complete disruption of the microtubules, the insulin-stimulated glucose transport activity was inhibited by 55%. This inhibition was concomitant with a comparable inhibition of GLUT4 translocation measured by the subcellular fractionation and the cell-surface GLUT4 labeling by trypsin cleavage. In addition, the time-course of insulin stimulation of the glucose transport activity was significantly delayed by microtubule disruption (t(1/2) were 7 and 2.3 min in nocodazole-treated and control cells, respectively), while the rate of GLUT4 endocytosis was little affected. The impaired insulin-stimulated glucose transport activity was not fully restored to the level in control cells by blocking GLUT4 endocytosis, suggesting that the inhibition was due to the existence of a microtubule-dependent subpopulation in the insulin-responsive GLUT4 pool. On the other hand, nocodazole partially inhibited insulin-induced translocation of the insulin-regulated aminopeptidase and the vesicle-associated membrane protein (VAMP)-2 without affecting GLUT1 and VAMP-3. In electrically permeabilized adipocytes, the insulin-stimulated glucose transport was inhibited by 40% by disruption of the microtubules whereas that stimulated with GTP gamma S was not affected. Intriguingly, the two reagents stimulated glucose transport to the comparable level by disruption of the microtubules. These data suggest that insulin recruits GLUT4 to the plasma membrane from at least two distinct intracellular compartments via distinct traffic routes with differential microtubule dependence in rat primary adipocytes. PMID- 12782633 TI - Pulmonary expression of leukemia inhibitory factor induces B cell hyperplasia and confers protection in hyperoxia. AB - Leukemia inhibitory factor (LIF) is produced by a large number of pulmonary cells in response to diverse stimuli. Exaggerated levels of LIF have also been detected in the adult respiratory distress syndrome and other disorders. The biologic effects of LIF in the lung, however, have not been elucidated. To define the respiratory effects of LIF, we generated transgenic mice in which human LIF was selectively targeted to the mature lung. In these mice, transgene activation caused an impressive increase in bronchoalveolar lavage (BAL) cellularity with a significant increase in BAL and tissue B lymphocytes. LIF also conferred protection in 100% O2 where it decreased alveolar-capillary protein leak and enhanced survival. This protective effect was associated with the induction of interleukin (IL)-6 mRNA and protein. LIF transgenic mice with a null mutation in IL-6 were more sensitive to the toxic effects of 100% O2 than LIF-transgenic animals with a wild-type IL-6 locus. These studies demonstrate that LIF induces B cell hyperplasia and confers protection in hyperoxic acute lung injury. They also demonstrate that LIF induces IL-6 and that the protective effects of LIF are mediated, in part, via this inductive event. LIF may be an important regulator of B cell-mediated responses and oxidant injury in the lung. PMID- 12782635 TI - Gts1p activates SNF1-dependent derepression of HSP104 and TPS1 in the stationary phase of yeast growth. AB - We previously reported that the GTS1 product, Gts1p, plays an important role in the regulation of heat tolerance of yeast under glucose-limited conditions in either batch or continuous culture. Here we show that heat tolerance was decreased in GTS1-deleted and increased in GTS1-overexpressing cells under glucose-derepressed conditions during the batch culture and that the disruption of SNF1, a transcriptional activator of glucose-repressible genes, diminished this effect of GTS1. Intracellular levels of Hsp104 and trehalose, which were reportedly required for the acquisition of heat tolerance in the stationary phase of cell growth, were affected in both GTS1 mutants roughly in proportion to the gene dosage of GTS1, whereas those of other Hsps were less affected. The mRNA levels of genes for Hsp104 and trehalose-6-phosphate synthase 1 changed as a function of GTS1 gene dosage. The Q-rich domain of Gts1p fused with the DNA binding domain of LexA activated the transcription of the reporter gene LacZ, and Gts1p lacking the Q-rich domain lost the activation activity of HSP104 and TPS1. Furthermore, Gts1p bound to subunits of Snf1 kinase, whereas it did not bind to DNA. Therefore, we suggested that GTS1 increases heat tolerance by mainly activating Snf1 kinase-dependent derepression of HSP104 and TPS1 in the stationary phase of yeast growth. PMID- 12782637 TI - Homogeneous hepatitis A virus particles. Proteolytic release of the assembly signal 2A from procapsids by factor Xa. AB - Among the picornaviridae, hepatitis A virus (HAV) is unique in that its assembly is driven by domain 2A of P1-2A, the precursor of the structural proteins (Probst, C., Jecht, M., and Gauss-Muller, V. (1999) J. Biol. Chem. 274, 4527 4531). Whereas infected individuals excrete in stool mature HAV capsids with VP1 as the major structural protein, its C-terminal extended form VP1-2A is the main component of immature procapsids produced in HAV-infected cells in culture. Obviously, a postassembly proteolytic step is required to remove the primary assembly signal 2A from VP1-2A of procapsids. Mutants of VP1-2A were expressed in COS7 cells to determine the cleavage site in VP1-2A and to test for the cleavage potential of viral and host proteinases (factor Xa and thrombin). Site-specific in vitro cleavage by factor Xa and thrombin occurred in procapsids that contained VP1-2A with engineered cognate cleavage sites for these proteinases. Interestingly, factor Xa but not thrombin liberated mature VP1 also from native procapsids in an assembly-dependent manner. The data show that domain 2A, which is required for pentamerization of its precursor polypeptides and thus for the primary step of HAV assembly, is removed from the surface of immature procapsid by a host proteinase. Moreover, our data open a novel avenue to produce homogeneous HAV particles from recombinant intermediates by in vitro treatment with exogenously added proteases such as factor Xa or thrombin. PMID- 12782636 TI - A serine protease inhibitor prevents endoplasmic reticulum stress-induced cleavage but not transport of the membrane-bound transcription factor ATF6. AB - Mammalian cells express several transcription factors embedded in the endoplasmic reticulum (ER) as transmembrane proteins that are activated by proteolysis, and two types of these proteins have been extensively investigated. One type comprises the sterol regulatory element-binding proteins (SREBP-1 and SREBP-2). The other type comprises the activating transcription factors 6 (ATF6alpha and ATF6beta), which are activated in response to ER stress. It was shown previously that both SREBP and ATF6 are cleaved sequentially first by the Site-1 protease (serine protease) and then by the Site-2 protease (metalloprotease) (Ye, J., Rawson, R. B., Komuro, R., Chen, X., Dave, U. P., Prywes, R., Brown, M. S., and Goldstein, J. L. (2000) Mol. Cell 6, 1355-1364). In this study, we examined various protease inhibitors and found that 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF), a serine protease inhibitor, prevented ER stress-induced cleavage of ATF6alpha and ATF6beta, resulting in inhibition of transcriptional induction of ATF6-target genes. AEBSF also inhibited production of the mature form of SREBP-2 that was induced in response to sterol depletion, and appeared to directly prevent cleavage of ATF6alpha and ATF6beta by inhibiting Site-1 protease. As the Site-1 protease is localized in the Golgi apparatus, both SREBP and ATF6 must relocate to the Golgi apparatus to be cleaved. We showed here that AEBSF treatment had little effect on ER stress-induced translocation of ATF6 from the ER to the Golgi apparatus, but blocked nuclear localization of ATF6. These results indicate that the transport of ATF6 from the ER to the Golgi apparatus and that from the Golgi apparatus to the nucleus are distinct steps that can be distinguished by treatment with AEBSF. PMID- 12782638 TI - Gene expression profiling reveals the mechanism and pathophysiology of mouse liver regeneration. AB - Comprehensive analysis of the changes in gene expression during liver regeneration was carried out by using an in-house microarray composed of 2,304 distinct mouse liver cDNA clones. Mice were subjected to partial two-thirds hepatectomy, and changes in mRNA levels were monitored up to 48 h. Of the 2,304 genes analyzed, 496 genes showed expression levels measurable at all time points after the partial hepatectomy. 317 genes were up- or down-regulated 2-fold or more at least at one time point during liver regeneration and were classified into eight clusters based on their expression patterns. With a more stringent cut off value of +/-2 S.D., 68 genes were listed and were classified into five clusters. In these two analyses with different clustering criteria, functionally categorized genes showed similar cluster distributions. Genes involved in protein synthesis and posttranslational processing were significantly enriched in the cluster characterized by rapid gene activation and subsequent persistence. This suggests the importance of modulating the efficiency of protein supply and/or altering the composition of protein population from the early phase of hepatocyte proliferation. Genes for two major liver functions, i.e. plasma protein secretion and intermediate metabolism were enriched in distinct clusters exhibiting the features of gradual gene activation and sustained repression, respectively. Therefore, these genes are differentially regulated during the regeneration, possibly leading to changes in the flow of amino acids and energy from enzyme proteins to plasma proteins in their synthesis. Thus, clustering analysis of expression patterns of functionally classified genes gave insights into mechanism and pathophysiology of liver regeneration. PMID- 12782639 TI - Mutant alpha-latrotoxin (LTXN4C) does not form pores and causes secretion by receptor stimulation: this action does not require neurexins. AB - Alpha-latrotoxin (LTX) causes massive release of neurotransmitters via a complex mechanism involving (i) activation of receptor(s) and (ii) toxin insertion into the plasma membrane with (iii) subsequent pore formation. Using cryo-electron microscopy, electrophysiological and biochemical methods, we demonstrate here that the recently described toxin mutant (LTXN4C) is unable to insert into membranes and form pores due to its inability to assemble into tetramers. However, this mutant still binds to major LTX receptors (latrophilin and neurexin) and causes strong transmitter exocytosis in synaptosomes, hippocampal slice cultures, neuromuscular junctions, and chromaffin cells. In the absence of mutant incorporation into the membrane, receptor activation must be the only mechanism by which LTXN4C triggers exocytosis. An interesting feature of this receptor-mediated transmitter release is its dependence on extracellular Ca2+. Because Ca2+ is also strictly required for LTX interaction with neurexin, the latter might be the only receptor mediating the LTXN4C action. To test this hypothesis, we used conditions (substitution of Ca2+ in the medium with Sr2+) under which LTXN4C does not bind to any member of the neurexin family but still interacts with latrophilin. We show that, in all the systems tested, Sr2+ fully replaces Ca2+ in supporting the stimulatory effect of LTXN4C. These results indicate that LTXN4C can cause neurotransmitter release just by stimulating a receptor and that neurexins are not critical for this receptor-mediated action. PMID- 12782640 TI - Identification of a novel transcription factor, GAGATA-binding protein, involved in androgen-mediated expression of prostate-specific antigen. AB - Prostate-specific antigen (PSA) is the most valuable marker for the evaluation of prostate cancer progression. The expression of PSA is controlled by androgen receptor (AR) through its binding to androgen-response elements (AREs). Several AREs have been identified within the 5.8-kb PSA promoter. The main activity of this 5.8-kb PSA promoter resides in a 455-bp enhancer core region located about 4 kb upstream of the TATA box. Our study suggests that in addition to the four AREs identified in the PSA enhancer core, another regulatory element (GAGATA), which is located at the region designated PSA3.1, also contributes to transcriptional regulation by androgens. Furthermore, electrophoretic mobility shift assay revealed that a putative transcriptional factor bound the GAGATA sequence in the PSA-producing prostate cancer cell. Further studies demonstrated that GAGATA factor preferentially bound the (G/C)(A/C/T)GATA sequence. The replacement of ATA with GGG in the GAGATA sequence completely eliminated the androgen-mediated transcriptional activity of the enhancer core. By using DNA-coupled magnetic beads and the Southwestern method, a 56-60-kDa protein was identified as the putative GAGATA binding factor. EMSA and Western blotting assay suggested that AR is not involved in androgen-mediated activation through PSA3.1. Therefore, we propose that binding of the GAGATA binding factor and AR to GAGATA and AREs, respectively, of the PSA enhancer core are required for the maximum transcriptional response to androgens. PMID- 12782641 TI - The tetraspanin CD151 functions as a negative regulator in the adhesion-dependent activation of Ras. AB - Transmembrane proteins of the tetraspanin superfamily are associated with integrins and are thought to regulate adhesion-dependent signaling. The molecular mechanisms of this regulation remain unknown. We used rat fibroblasts to analyze the contribution of the tetraspanin CD151 in the adhesion-dependent signaling. Expression of CD151 specifically attenuated adhesion-dependent activation of Ras. Furthermore, activation of PKB/c-Akt and ERK1/2, downstream targets in the Ras signaling pathway, was also diminished in cells expressing CD151. In contrast, adhesion-dependent activation of FAK and c-Src were not affected by CD151. The attenuation of Ras signaling did not correlate with phosphorylation of Tyr925 FAK, tyrosine phosphorylation of Shc, or with assembly of the p120RasGAP-p62Dok complex. Using mutants of CD151 we established that the cytoplasmic C-terminal portion is critical for activity of CD151 toward Ras. Taken together these results identify CD151 as a negative regulator of Ras and suggest a novel mechanism of adhesion-dependent regulation of Ras activity. PMID- 12782642 TI - Increased renal medullary H2O2 leads to hypertension. AB - We have recently reported that exaggerated oxidative stress in the renal medulla due to superoxide dismutase inhibition resulted in a reduction of renal medullary blood flow and sustained hypertension. The present study tested the hypothesis that selective scavenging of O2*- in the renal medulla would prevent hypertension associated with this exaggerated oxidative stress. An indwelling, aortic catheter was implanted in nonnephrectomized Sprague-Dawley rats for daily measurement of arterial blood pressure, and a renal medullary interstitial catheter was implanted for continuous delivery of the superoxide dismutase inhibitor diethyldithiocarbamic acid (DETC, 7.5 mg x kg(-1) x d(-1)) and a chemical superoxide dismutase mimetic, 4-hydroxytetramethyl piperidine-1-oxyl (TEMPOL, 10 mg. kg-1. d-1). Renal medullary interstitial infusion of TEMPOL completely blocked DETC-induced accumulation of O2*- in the renal medulla, as measured by the conversion rate of dihydroethidium to ethidium in the dialysate and by urinary excretion of 8-isoprostanes. However, TEMPOL infusion failed to prevent DETC-induced hypertension, unless catalase (5 mg x kg(-1) d(-1)) was coinfused. Direct infusion of H2O2 into the renal medulla resulted in increases of mean arterial pressure from 115+/-2.5 to 131+/-2.1 mm Hg, which was similar to that observed in rats receiving the medullary infusion of both TEMPOL and DETC. The results indicate that sufficient catalase activity in the renal medulla is a prerequisite for the antihypertensive action of TEMPOL and that accumulated H2O2 in the renal medulla associated with exaggerated oxidative stress might have a hypertensive consequence. PMID- 12782643 TI - Pharmacological modulation of platelet function in hypertension. AB - Platelets exert a considerable influence on human morbidity and mortality. The rationale for their study in hypertension follows the observation that the major consequences of hypertension are stroke and myocardial infarction. However, the etiology of these consequences in hypertension is, paradoxically, not hemorrhagic (as might be expected from the effects of high blood pressure), but occlusive, with thrombus being the culprit lesion. Mechanisms of platelet activation include high shear force, activation of the renin-angiotensin system, endothelial changes, and the presence of comorbidity, such as atrial fibrillation. The treatment of high blood pressure brings about a reversal of the changes seen in the cell. This could be in part due to the direct effect of the drug on the megakaryocyte and/or the platelets themselves, or it might simply be due to the reduction in blood pressure. Some drugs, such as calcium channel antagonists and angiotensin II receptor blockers, however, might have direct effects on platelet biochemistry other than reducing blood pressure. Finally, antiplatelet drugs are becoming an important part of the management of high-risk hypertensives, which aim to minimize vascular complications. PMID- 12782644 TI - Baroreflex failure as a late sequela of neck irradiation. AB - Combined chemotherapy and radiotherapy increase long-term survival in patients with head and neck tumors. Late complications of treatment, however, are being recognized increasingly. Surgery or radiotherapy of the carotid sinuses or brain stem can evoke labile hypertension and orthostatic intolerance from acute or subacute baroreflex failure. Here we report cases in which chronic baroreflex failure appeared to develop as a late sequela of neck irradiation. Three patients referred for autonomic nervous system function testing had labile blood pressure and chronic orthostatic intolerance that developed years after neck irradiation for cancer. In each patient, heart rate remained constant during performance of the Valsalva maneuver, suggesting baroreflex-cardiovagal failure. All 3 patients had virtually zero baroreflex-cardiovagal gain, quantified by interbeat interval systolic blood pressure relationships after intravenous phenylephrine or nitroglycerine. Ambulatory blood pressure monitoring revealed highly variable blood pressure, with sudden pressor and depressor episodes, a characteristic feature of baroreflex failure. Cardiovagal efferent function, assessed by power spectral analysis of heart rate variability during slow, deep respiration, was normal. Sympathetic noradrenergic efferent function, assessed by cold pressor testing and plasma catecholamine levels during supine rest and orthostasis, was also normal or increased. These findings indicated a primarily afferent lesion. Carotid ultrasonography revealed intimal thickening and atheromatous plaques in all 3 patients. We propose that labile hypertension and orthostatic intolerance can develop as a late sequela of neck irradiation, due to chronic carotid baroreflex failure, which in turn is due to radiation-induced accelerated development of carotid arteriosclerosis. Splinting of carotid sinus mechanoreceptors in rigidified arterial walls would impede detection of alterations in blood pressure and thereby disrupt baroreflex regulation of cardiovagal and sympathetic outflows. PMID- 12782645 TI - Endothelin antagonism on aldosterone-induced oxidative stress and vascular remodeling. AB - Endothelin A (ETA) receptor blockade has prevented vascular remodeling in aldosterone and salt-induced hypertension. To evaluate effects of the ETA receptor antagonist, BMS 182874, compared with the aldosterone antagonist, spironolactone, on vascular remodeling in aldosterone-infused rats not exposed to a high salt diet, Sprague-Dawley rats were infused subcutaneously with aldosterone (0.75 microg/h) and treated with BMS 182874 (40 mg. kg-1. d-1), spironolactone, or hydralazine (both 25 mg. kg-1. d-1) while receiving a normal salt diet for 6 weeks. Aldosterone increased systolic BP (P<0.01), plasma endothelin (3.33+/-0.32 versus 1.85+/-0.40 pmol/L in control, P<0.05), systemic oxidative stress as shown by plasma thiobarbituric acid-reacting substances and vascular nicotinamide adenine dinucleotide phosphate (NADPH) activity. Aldosterone increased small artery media thickness (17.7+/-0.9 versus 13.6+/-0.8 microm in control, P<0.05) and media/lumen ratio (7.6+/-0.4 versus 5.5+/-0.4% in control, P<0.05), with growth index of 21% indicating hypertrophic remodeling. Laser confocal microscopy showed increased collagen and fibronectin deposition and intercellular adhesion molecule-1 (ICAM-1) content in the vessel wall of aldosterone-infused rats. The 3 treatments lowered BP, although hydralazine was slightly less effective. BMS 182874 and spironolactone decreased oxidative stress, normalized the hypertrophic remodeling, decreased collagen and fibronectin deposition, and reduced ICAM-1 abundance in the vascular wall of aldosterone-infused rats, whereas hydralazine only reduced NADPH activity in aorta but did not affect the remaining parameters. Vascular remodeling of small arteries occurs in aldosterone-infused rats exposed to a normal salt diet and may be mediated in part by ET-1 via stimulation of ETA receptors. Endothelin blockade may exert beneficial effects on vascular remodeling, fibrosis, oxidative stress, and adhesion molecule expression in aldosterone-induced hypertension. PMID- 12782648 TI - Diversified transcription initiation complexes expand promoter selectivity and tissue-specific gene expression. PMID- 12782649 TI - Strange bedfellows: polyadenylation factors at the promoter. PMID- 12782650 TI - TRF1 is degraded by ubiquitin-mediated proteolysis after release from telomeres. AB - Mammalian telomeres are coated by the sequence-specific, DNA-binding protein, TRF1, a negative regulator of telomere length. Previous results showed that ADP ribosylation of TRF1 by tankyrase 1 released TRF1 from telomeres and promoted telomere elongation. We now show that loss of TRF1 from telomeres results in ubiquitination and degradation of TRF1 by the proteasome and that degradation is required to keep TRF1 off telomeres. Ubiquitination of TRF1 is regulated by its telomere-binding status; only the telomere-unbound form of TRF1 is ubiquitinated. Our findings suggest a novel mechanism of sequential post translational modification of TRF1 (ADP-ribosylation and ubiquitination) for regulating access of telomerase to telomeres. PMID- 12782651 TI - Local spreading of MSL complexes from roX genes on the Drosophila X chromosome. AB - MSL proteins and noncoding roX RNAs form complexes to up-regulate hundreds of genes on the Drosophila male X chromosome, and make X-linked gene expression equal in males and females. Altering the ratio of MSL proteins to roX RNA dramatically changes X-chromosome morphology. In protein excess, the MSL complex concentrates near sites of roX transcription and is depleted elsewhere. These results support a model for distribution of MSL complexes, in which local spreading in cis from roX genes is balanced with diffusion of soluble complexes in trans. When overexpressed, MSL proteins can recognize the X chromosome, modify histones, and partially restore male viability even in the absence of roX RNAs. Thus, the protein components can carry out all essential functions of dosage compensation, but roX RNAs facilitate the correct targeting of MSL complexes, in part by nucleation of spreading from their sites of synthesis. PMID- 12782652 TI - Generation of Ski-knockdown mice by expressing a long double-strand RNA from an RNA polymerase II promoter. AB - We have developed a new vector, named pDECAP, to express long double-strand RNA (ds-RNA) from an RNA polymerase II (Pol II) promoter. Because the transcripts from pDECAP lack both the 5'-cap structure and the 3'-poly(A) tail that facilitate ds-RNA export to the cytoplasm, long ds-RNA from pDECAP does not induce the interferon response. Transgenic mice embryos expressing long ds-RNA for the transcriptional corepressor Ski from this vector exhibited phenotypes that were remarkably similar to those of Ski-deficient embryos, including defects of neural tube closure and eye formation. Thus, this vector provides a new tool to efficiently generate tissue-specific knockdown mice for studying gene function in whole animal systems. PMID- 12782653 TI - phospholipase C gamma-1 is required downstream of vascular endothelial growth factor during arterial development. AB - In this study, we utilize transgenic zebrafish with fluorescently labeled blood vessels to identify and characterize a mutant (y10) that displays specific defects in the formation of arteries, but not veins. We find that y10 encodes phospholipase C gamma-1 (plcg1), a known effector of receptor tyrosine kinase signaling. We further show that plcg1y10 mutant embryos fail to respond to exogenous Vegf. Our results indicate that Plcg1 functions specifically downstream of the Vegf receptor during embryonic development to govern formation of the arterial system. PMID- 12782654 TI - Dwarfism, impaired skin development, skeletal muscle atrophy, delayed bone development, and impeded adipogenesis in mice lacking Akt1 and Akt2. AB - To elucidate the functions of the serine/threonine kinase Akt/PKB in vivo, we generated mice lacking both akt1 and akt2 genes. Akt1/Akt2 double-knockout (DKO) mice exhibit severe growth deficiency and die shortly after birth. These mice display impaired skin development because of a proliferation defect, severe skeletal muscle atrophy because of a marked decrease in individual muscle cell size, and impaired bone development. These defects are strikingly similar to the phenotypes of IGF-1 receptor-deficient mice and suggest that Akt may serve as the most critical downstream effector of the IGF-1 receptor during development. In addition, Akt1/Akt2 DKO mice display impeded adipogenesis. Specifically, Akt1 and Akt2 are required for the induced expression of PPARgamma, the master regulator of adipogenesis, establishing a new essential role for Akt in adipocyte differentiation. Overall, the combined deletion of Akt1 and Akt2 establishes in vivo roles for Akt in cell proliferation, growth, and differentiation. These functions of Akt were uncovered despite the observed lower level of Akt activity mediated by Akt3 in Akt1/Akt2 DKO cells, suggesting that a critical threshold level of Akt activity is required to maintain normal cell proliferation, growth, and differentiation. PMID- 12782655 TI - Loss of circadian rhythmicity in aging mPer1-/-mCry2-/- mutant mice. AB - The mPer1, mPer2, mCry1, and mCry2 genes play a central role in the molecular mechanism driving the central pacemaker of the mammalian circadian clock, located in the suprachiasmatic nuclei (SCN) of the hypothalamus. In vitro studies suggest a close interaction of all mPER and mCRY proteins. We investigated mPER and mCRY interactions in vivo by generating different combinations of mPer/mCry double mutant mice. We previously showed that mCry2 acts as a nonallelic suppressor of mPer2 in the core clock mechanism. Here, we focus on the circadian phenotypes of mPer1/mCry double-mutant animals and find a decay of the clock with age in mPer1 /- mCry2-/- mice at the behavioral and the molecular levels. Our findings indicate that complexes consisting of different combinations of mPER and mCRY proteins are not redundant in vivo and have different potentials in transcriptional regulation in the system of autoregulatory feedback loops driving the circadian clock. PMID- 12782656 TI - The POU proteins Brn-2 and Oct-6 share important functions in Schwann cell development. AB - The genetic hierarchy that controls myelination of peripheral nerves by Schwann cells includes the POU domain Oct-6/Scip/Tst-1and the zinc-finger Krox-20/Egr2 transcription factors. These pivotal transcription factors act to control the onset of myelination during development and tissue regeneration in adults following damage. In this report we demonstrate the involvement of a third transcription factor, the POU domain factor Brn-2. We show that Schwann cells express Brn-2 in a developmental profile similar to that of Oct-6 and that Brn-2 gene activation does not depend on Oct-6. Overexpression of Brn-2 in Oct-6 deficient Schwann cells, under control of the Oct-6 Schwann cell enhancer (SCE), results in partial rescue of the developmental delay phenotype, whereas compound disruption of both Brn-2 and Oct-6 results in a much more severe phenotype. Together these data strongly indicate that Brn-2 function largely overlaps with that of Oct-6 in driving the transition from promyelinating to myelinating Schwann cells. PMID- 12782657 TI - Localized recruitment of a chromatin-remodeling activity by an activator in vivo drives transcriptional elongation. AB - To understand the role of chromatin-remodeling activities in transcription, it is necessary to understand how they interact with transcriptional activators in vivo to regulate the different steps of transcription. Human heat shock factor 1 (HSF1) stimulates both transcriptional initiation and elongation. We replaced mouse HSF1 in fibroblasts with wild-type and mutant human HSF1 constructs and characterized regulation of an endogenous mouse hsp70 gene. A mutation that diminished transcriptional initiation led to twofold reductions in hsp70 mRNA induction and recruitment of a SWI/SNF remodeling complex. In contrast, a mutation that diminished transcriptional elongation abolished induction of full length mRNA, SWI/SNF recruitment, and chromatin remodeling, but minimally impaired initiation from the hsp70 promoter. Another remodeling factor, SNF2h, is constitutively present at the promoter irrespective of the genotype of HSF1. These data suggest that localized recruitment of SWI/SNF drives a specialized remodeling reaction necessary for the production of full-length hsp70 mRNA. PMID- 12782660 TI - SARS coronavirus: a new challenge for prevention and therapy. PMID- 12782659 TI - Yeast enhancer of polycomb defines global Esa1-dependent acetylation of chromatin. AB - Drosophila Enhancer of Polycomb, E(Pc), is a suppressor of position-effect variegation and an enhancer of both Polycomb and trithorax mutations. A homologous yeast protein, Epl1, is a subunit of the NuA4 histone acetyltransferase complex. Epl1 depletion causes cells to accumulate in G2/M and global loss of acetylated histones H4 and H2A. In relation to the Drosophila protein, mutation of Epl1 suppresses gene silencing by telomere position effect. Epl1 protein is found in the NuA4 complex and a novel highly active smaller complex named Piccolo NuA4 (picNuA4). The picNuA4 complex contains Esa1, Epl1, and Yng2 as subunits and strongly prefers chromatin over free histones as substrate. Epl1 conserved N-terminal domain bridges Esa1 and Yng2 together, stimulating Esa1 catalytic activity and enabling acetylation of chromatin substrates. A recombinant picNuA4 complex shows characteristics similar to the native complex, including strong chromatin preference. Cells expressing only the N-terminal half of Epl1 lack NuA4 HAT activity, but possess picNuA4 complex and activity. These results indicate that the essential aspect of Esa1 and Epl1 resides in picNuA4 function. We propose that picNuA4 represents a nontargeted histone H4/H2A acetyltransferase activity responsible for global acetylation, whereas the NuA4 complex is recruited to specific genomic loci to perturb locally the dynamic acetylation/deacetylation equilibrium. PMID- 12782658 TI - NELF and DSIF cause promoter proximal pausing on the hsp70 promoter in Drosophila. AB - NELF and DSIF collaborate to inhibit elongation by RNA polymerase IIa in extracts from human cells. A multifaceted approach was taken to investigate the potential role of these factors in promoter proximal pausing on the hsp70 gene in Drosophila. Immunodepletion of DSIF from a Drosophila nuclear extract reduced the level of polymerase that paused in the promoter proximal region of hsp70. Depletion of one NELF subunit in salivary glands using RNA interference also reduced the level of paused polymerase. In vivo protein-DNA cross-linking showed that NELF and DSIF associate with the promoter region before heat shock. Immunofluorescence analysis of polytene chromosomes corroborated the cross linking result and showed that NELF, DSIF, and RNA polymerase IIa colocalize at the hsp70 genes, small heat shock genes, and many other chromosomal locations. Finally, following heat shock induction, DSIF and polymerase but not NELF were strongly recruited to chromosomal puffs harboring the hsp70 genes. We propose that NELF and DSIF cause polymerase to pause in the promoter proximal region of hsp70. The transcriptional activator, HSF, might cause NELF to dissociate from the elongation complex. DSIF continues to associate with the elongation complex and could serve a positive role in elongation. PMID- 12782661 TI - CT screening: a trade-off of risks, benefits, and costs. PMID- 12782662 TI - Molecular-genetic imaging: current and future perspectives. PMID- 12782664 TI - A new hypothesis for how sex steroid hormones regulate bone mass. AB - It has been proposed - but remains controversial - that estrogen's effects on various tissues may be mediated by different cell signaling pathways. Researchers have identified a synthetic ligand that activates only a subset of these pathways, suggesting that bypass of the traditional estrogen pathway can prevent bone loss without associated side effects on reproductive organs. PMID- 12782665 TI - What is the real role of antimicrobial polypeptides that can mediate several other inflammatory responses? AB - Antimicrobial peptides are effector molecules of innate immunity with microbicidal and pro- or anti-inflammatory activities. Their role is now widening following evidence that one such multifunctional peptide, LL-37, induces angiogenesis, a process essential for host defense, wound healing, and tissue repair. PMID- 12782663 TI - Cardiovascular nuclear magnetic resonance: basic and clinical applications. PMID- 12782666 TI - Functionally mature virus-specific CD8(+) T memory cells in congenitally infected newborns: proof of principle for neonatal vaccination? AB - The presence in newborns of a mature and functional CD8(+) T cell response to congenital cytomegalovirus infection suggests that the machinery necessary to prime such responses is present in utero and raises questions related to neonatal vaccination. PMID- 12782667 TI - New insights into a persistent problem -- chlamydial infections. AB - Tissue tropism of clinical ocular and genital Chlamydia trachomatis strains is shown to be linked to the tryptophan synthase genotype. It is suggested that, in the presence of IFN-gamma, which depletes available tryptophan, there exist unique host-parasite interactions that may contribute to persistent chlamydial infection. PMID- 12782669 TI - An angiogenic role for the human peptide antibiotic LL-37/hCAP-18. AB - Antimicrobial peptides are effector molecules of the innate immune system and contribute to host defense and regulation of inflammation. The human cathelicidin antimicrobial peptide LL-37/hCAP-18 is expressed in leukocytes and epithelial cells and secreted into wound and airway surface fluid. Here we show that LL-37 induces angiogenesis mediated by formyl peptide receptor-like 1 expressed on endothelial cells. Application of LL-37 resulted in neovascularization in the chorioallantoic membrane assay and in a rabbit model of hind-limb ischemia. The peptide directly activates endothelial cells, resulting in increased proliferation and formation of vessel-like structures in cultivated endothelial cells. Decreased vascularization during wound repair in mice deficient for CRAMP, the murine homologue of LL-37/hCAP-18, shows that cathelicidin-mediated angiogenesis is important for cutaneous wound neovascularization in vivo. Taken together, these findings demonstrate that LL-37/hCAP-18 is a multifunctional antimicrobial peptide with a central role in innate immunity by linking host defense and inflammation with angiogenesis and arteriogenesis. PMID- 12782668 TI - Kinase-mediated regulation of common transcription factors accounts for the bone protective effects of sex steroids. AB - It has been found that 4-estren-3alpha,17beta-diol, a synthetic ligand for the estrogen receptor (ER) or androgen receptor (AR), which does not affect classical transcription, reverses bone loss in ovariectomized females or orchidectomized males without affecting the uterus or seminal vesicles, demonstrating that the classical genotropic actions of sex steroid receptors are dispensable for their bone-protective effects, but indispensable for their effects on reproductive organs. We have now investigated the mechanism of action of this compound. We report that, identically to 17beta-estradiol or dihydrotestosterone, but differently from raloxifene, estren alters the activity of Elk-1, CCAAT enhancer binding protein-beta (C/EBPbeta), and cyclic adenosine monophosphate-response element binding protein (CREB), or c-Jun/c-Fos by an extranuclear action of the ER or AR, resulting in activation of the Src/Shc/ERK pathway or downregulation of JNK, respectively. All of these effects are non-sex specific, require only the ligand-binding domain of the receptor, and are indispensable for the antiapoptotic action of these ligands on osteoblastic and HeLa cells. Moreover, administration of 17beta-estradiol or 4-estren-3alpha,17beta-diol to ovariectomized mice induces phosphorylation of ERKs, Elk-1, and C/EBPbeta, downregulates c-Jun, and upregulates the expression of egr-1, an ERK/SRE target gene. Kinase-initiated regulation of commonly used transcription factors offers a molecular explanation for the profound skeletal effects of sex steroid receptor ligands, including synthetic ones that are devoid of classical transcriptional activity. PMID- 12782670 TI - In vivo administration of a lentiviral vaccine targets DCs and induces efficient CD8(+) T cell responses. AB - The present study evaluates the potential of third-generation lentivirus vectors with respect to their use as in vivo-administered T cell vaccines. We demonstrate that lentivector injection into the footpad of mice transduces DCs that appear in the draining lymph node and in the spleen. In addition, a lentivector vaccine bearing a T cell antigen induced very strong systemic antigen-specific cytotoxic T lymphocyte (CTL) responses in mice. Comparative vaccination performed in two different antigen models demonstrated that in vivo administration of lentivector was superior to transfer of transduced DCs or peptide/adjuvant vaccination in terms of both amplitude and longevity of the CTL response. Our data suggest that a decisive factor for efficient T cell priming by lentivector might be the targeting of DCs in situ and their subsequent migration to secondary lymphoid organs. The combination of performance, ease of application, and absence of pre existing immunity in humans make lentivector-based vaccines an attractive candidate for cancer immunotherapy. PMID- 12782671 TI - Mice deficient in alpha-actinin-4 have severe glomerular disease. AB - Dominantly inherited mutations in ACTN4, which encodes alpha-actinin-4, cause a form of human focal and segmental glomerulosclerosis (FSGS). By homologous recombination in ES cells, we developed a mouse model deficient in Actn4. Mice homozygous for the targeted allele have no detectable alpha-actinin-4 protein expression. The number of homozygous mice observed was lower than expected under mendelian inheritance. Surviving mice homozygous for the targeted allele show progressive proteinuria, glomerular disease, and typically death by several months of age. Light microscopic analysis shows extensive glomerular disease and proteinaceous casts. Electron microscopic examination shows focal areas of podocyte foot-process effacement in young mice, and diffuse effacement and globally disrupted podocyte morphology in older mice. Despite the widespread distribution of alpha-actinin-4, histologic examination of mice showed abnormalities only in the kidneys. In contrast to the dominantly inherited human form of ACTN4-associated FSGS, here we show that the absence of alpha-actinin-4 causes a recessive form of disease in mice. Cell motility, as measured by lymphocyte chemotaxis assays, was increased in the absence of alpha-actinin-4. We conclude that alpha-actinin-4 is required for normal glomerular function. We further conclude that the nonsarcomeric forms of alpha-actinin (alpha-actinin-1 and alpha-actinin-4) are not functionally redundant. In addition, these genetic studies demonstrate that the nonsarcomeric alpha-actinin-4 is involved in the regulation of cell movement. PMID- 12782672 TI - Alpha1-adrenergic receptors activate Ca(2+)-permeable cationic channels in prostate cancer epithelial cells. AB - The prostate gland is a rich source of alpha1-adrenergic receptors (alpha1-ARs). alpha1-AR antagonists are commonly used in the treatment of benign prostatic hyperplasia symptoms, due to their action on smooth muscle cells. However, virtually nothing is known about the role of alpha1-ARs in epithelial cells. Here, by using two human prostate cancer epithelial (hPCE) cell models - primary cells from resection specimens (primary hPCE cells) and an LNCaP (lymph node carcinoma of the prostate) cell line - we identify an alpha1A subtype of adrenergic receptor (alpha1A-AR) and show its functional coupling to plasmalemmal cationic channels via direct diacylglycerol (DAG) gating. In both cell types, agonist-mediated stimulation of alpha1A-ARs and DAG analogues activated similar cationic membrane currents and Ca(2+) influx. These currents were sensitive to the alpha1A-AR antagonists, prazosin and WB4101, and to transient receptor potential (TRP) channel blockers, 2-aminophenyl borate and SK&F 96365. Chronic activation of alpha1A-ARs enhanced LNCaP cell proliferation, which could be antagonized by alpha1A-AR and TRP inhibitors. Collectively, our results suggest that alpha1-ARs play a role in promoting hPCE cell proliferation via TRP channels. PMID- 12782673 TI - The voltage-gated Kv1.3 K(+) channel in effector memory T cells as new target for MS. AB - Through a combination of fluorescence microscopy and patch-clamp analysis we have identified a striking alteration in K(+) channel expression in terminally differentiated human CCR7(-)CD45RA(-) effector memory T lymphocytes (T(EM)). Following activation, T(EM) cells expressed significantly higher levels of the voltage-gated K(+) channel Kv1.3 and lower levels of the calcium-activated K(+) channel IKCa1 than naive and central memory T cells (T(CM)). Upon repeated in vitro antigenic stimulation, naive cells differentiated into Kv1.3(high)IKCa1(low) T(EM) cells, and the potent Kv1.3-blocking sea anemone Stichodactyla helianthus peptide (ShK) suppressed proliferation of T(EM) cells without affecting naive or T(CM) lymphocytes. Thus, the Kv1.3(high)IKCa1(low) phenotype is a functional marker of activated T(EM) lymphocytes. Activated myelin reactive T cells from patients with MS exhibited the Kv1.3(high)IKCa1(low) T(EM) phenotype, suggesting that they have undergone repeated stimulation during the course of disease; these cells may contribute to disease pathogenesis due to their ability to home to inflamed tissues and exhibit immediate effector function. The Kv1.3(high)IKCa1(low) phenotype was not seen in glutamic acid decarboxylase, insulin-peptide or ovalbumin-specific and mitogen-activated T cells from MS patients, or in myelin-specific T cells from healthy controls. Selective targeting of Kv1.3 in T(EM) cells may therefore hold therapeutic promise for MS and other T cell-mediated autoimmune diseases. PMID- 12782674 TI - Obesity resistance and enhanced glucose metabolism in mice transplanted with white adipose tissue lacking acyl CoA:diacylglycerol acyltransferase 1. AB - Recent studies have identified the white adipose tissue (WAT) as an important endocrine organ that regulates energy and glucose metabolism via a number of secreted factors. Mice lacking acyl CoA:diacylglycerol acyltransferase 1 (DGAT1), a key enzyme in mammalian triglyceride synthesis, are protected against diet induced obesity and glucose intolerance because of increased energy expenditure and enhanced insulin sensitivity. Because DGAT1 is highly expressed in WAT, we hypothesized that DGAT1 deficiency affects the expression of adipocyte-derived factors that regulate energy and glucose metabolism. Here we show that the transplantation of DGAT1-deficient WAT decreases adiposity and enhances glucose disposal in wild-type mice. Analysis of DGAT1-deficient WAT revealed a twofold increase in the expression of adiponectin, a molecule that enhances fatty acid oxidation and insulin sensitivity, and this increase may account in part for the transplantation-induced metabolic changes. Our results highlight the importance of the endocrine function of WAT and suggest that an alteration in this function contributes to the increased energy expenditure and insulin sensitivity in DGAT1 deficient mice. PMID- 12782675 TI - The secretory proprotein convertases furin, PC5, and PC7 activate VEGF-C to induce tumorigenesis. AB - The secretory factor VEGF-C has been directly implicated in various physiological processes during embryogenesis and human cancers. However, the importance of the conversion of its precursor proVEGF-C to mature VEGF-C in tumorigenesis, and vessel formation and the identity of the protease(s) that regulate these processes is/are not known. The intracellular processing of proVEGF-C that occurs within the dibasic motif HSIIRR(227)SL suggests the involvement of the proprotein convertases (PCs) in this process. In addition, furin and VEGF-C were found to be coordinately expressed in adult mouse tissues. Cotransfection of the furin deficient colon carcinoma cell line LoVo with proVEGF-C and different PC members revealed that furin, PC5, and PC7 are candidate VEGF-C convertases. This finding is consistent with the in vitro digestions of an internally quenched synthetic fluorogenic peptide mimicking the cleavage site of proVEGF-C ((220)Q-VHSIIRR downward arrow SLP(230)). The processing of proVEGF-C is blocked by the inhibitory prosegments of furin, PC5, and PACE4, as well as by furin-motif variants of alpha2-macroglobulin and alpha1-antitrypsin. Subcutaneous injection of CHO cells stably expressing VEGF-C into nude mice enhanced angiogenesis and lymphangiogenesis, but not tumor growth. In contrast, expression of proVEGF-C obtained following mutation of the cleavage site (HSIIRR(227)SL to HSIISS(227)SL) inhibits angiogenesis and lymphangiogenesis as well as tumor growth. Our findings demonstrate the processing of proVEGF-C by PCs and highlight the potential use of PC inhibitors as agents for inhibiting malignancies induced by VEGF-C. PMID- 12782676 TI - Thyroid functions of mouse cathepsins B, K, and L. AB - Thyroid function depends on processing of the prohormone thyroglobulin by sequential proteolytic events. From in vitro analysis it is known that cysteine proteinases mediate proteolytic processing of thyroglobulin. Here, we have analyzed mice with deficiencies in cathepsins B, K, L, B and K, or K and L in order to investigate which of the cysteine proteinases is most important for proteolytic processing of thyroglobulin in vivo. Immunolabeling demonstrated a rearrangement of the endocytic system and a redistribution of extracellularly located enzymes in thyroids of cathepsin-deficient mice. Cathepsin L was upregulated in thyroids of cathepsin K(-/-) or B(-/-)/K(-/-) mice, suggesting a compensation of cathepsin L for cathepsin K deficiency. Impaired proteolysis resulted in the persistence of thyroglobulin in the thyroids of mice with deficiencies in cathepsin B or L. The typical multilayered appearance of extracellularly stored thyroglobulin was retained in cathepsin K(-/-) mice only. These results suggest that cathepsins B and L are involved in the solubilization of thyroglobulin from its covalently cross-linked storage form. Cathepsin K(-/ )/L(-/-) mice had significantly reduced levels of free thyroxine, indicating that utilization of luminal thyroglobulin for thyroxine liberation is mediated by a combinatory action of cathepsins K and L. PMID- 12782677 TI - Mature CD8(+) T lymphocyte response to viral infection during fetal life. AB - Immunization of newborns against viral infections may be hampered by ineffective CD8(+) T cell responses. To characterize the function of CD8(+) T lymphocytes in early life, we studied newborns with congenital human cytomegalovirus (HCMV) infection. We demonstrate that HCMV infection in utero leads to the expansion and the differentiation of mature HCMV-specific CD8(+) T cells, which have similar characteristics to those detected in adults. High frequencies of HCMV-specific CD8(+) T cells were detected by ex vivo tetramer staining as early as after 28 weeks of gestation. During the acute phase of infection, these cells had an early differentiation phenotype (CD28(-)CD27(+)CD45RO(+), perforin(low)), and they acquired a late differentiation phenotype (CD28(-)CD27(-)CD45RA(+), perforin(high)) during the course of the infection. The differentiated cells showed potent perforin-dependent cytolytic activity and produced antiviral cytokines. The finding of a mature and functional CD8(+) T cell response to HCMV suggests that the machinery required to prime such responses is in place during fetal life and could be used to immunize newborns against viral pathogens. PMID- 12782679 TI - Selective inhibitors of the osteoblast proteasome stimulate bone formation in vivo and in vitro. AB - We have found that the ubiquitin-proteasome pathway exerts exquisite control of osteoblast differentiation and bone formation in vitro and in vivo in rodents. Structurally different inhibitors that bind to specific catalytic beta subunits of the 20S proteasome stimulated bone formation in bone organ cultures in concentrations as low as 10 nM. When administered systemically to mice, the proteasome inhibitors epoxomicin and proteasome inhibitor-1 increased bone volume and bone formation rates over 70% after only 5 days of treatment. Since the ubiquitin-proteasome pathway has been shown to modulate expression of the Drosophila homologue of the bone morphogenetic protein-2 and -4 (BMP-2 and BMP-4) genes, we examined the effects of noggin, an endogenous inhibitor of BMP-2 and BMP-4 on bone formation stimulated by these compounds and found that it was abrogated. These compounds increased BMP-2 but not BMP-4 or BMP-6 mRNA expression in osteoblastic cells, suggesting that BMP-2 was responsible for the observed bone formation that was inhibited by noggin. We show proteasome inhibitors regulate BMP-2 gene expression at least in part through inhibiting the proteolytic processing of Gli3 protein. Our results suggest that the ubiquitin proteasome machinery regulates osteoblast differentiation and bone formation and that inhibition of specific components of this system may be useful therapeutically in common diseases of bone loss. PMID- 12782678 TI - Polymorphisms in Chlamydia trachomatis tryptophan synthase genes differentiate between genital and ocular isolates. AB - We previously reported that laboratory reference strains of Chlamydia trachomatis differing in infection organotropism correlated with inactivating mutations in the pathogen's tryptophan synthase (trpBA) genes. Here, we have applied functional genomics to extend this work and find that the paradigm established for reference serovars also applies to clinical isolates - specifically, all ocular trachoma isolates tested have inactivating mutations in the synthase, whereas all genital isolates encode a functional enzyme. Moreover, functional enzyme activity was directly correlated to IFN-gamma resistance through an indole rescue mechanism. Hence, a strong selective pressure exists for genital strains to maintain a functional synthase capable of using indole for tryptophan biosynthesis. The fact that ocular serovars (serovar B) isolated from the genital tract were found to possess a functional synthase provided further persuasive evidence of this association. These results argue that there is an important host parasite relationship between chlamydial genital strains and the human host that determines organotropism of infection and the pathophysiology of disease. We speculate that this relationship involves the production of indole by components of the vaginal microbial flora, allowing chlamydiae to escape IFN-gamma-mediated eradication and thus establish persistent infection. PMID- 12782680 TI - The alpha(1A/C)- and alpha(1B)-adrenergic receptors are required for physiological cardiac hypertrophy in the double-knockout mouse. AB - Catecholamines and alpha(1)-adrenergic receptors (alpha(1)-ARs) cause cardiac hypertrophy in cultured myocytes and transgenic mice, but heart size is normal in single KOs of the main alpha(1)-AR subtypes, alpha(1A/C) and alpha(1B). Here we tested whether alpha(1)-ARs are required for developmental cardiac hypertrophy by generating alpha(1A/C) and alpha(1B) double KO (ABKO) mice, which had no cardiac alpha(1)-AR binding. In male ABKO mice, heart growth after weaning was 40% less than in WT, and the smaller heart was due to smaller myocytes. Body and other organ weights were unchanged, indicating a specific effect on the heart. Blood pressure in ABKO mice was the same as in WT, showing that the smaller heart was not due to decreased load. Contractile function was normal by echocardiography in awake mice, but the smaller heart and a slower heart rate reduced cardiac output. alpha(1)-AR stimulation did not activate extracellular signal-regulated kinase (Erk) and downstream kinases in ABKO myocytes, and basal Erk activity was lower in the intact ABKO heart. In female ABKO mice, heart size was normal, even after ovariectomy. Male ABKO mice had reduced exercise capacity and increased mortality with pressure overload. Thus, alpha(1)-ARs in male mice are required for the physiological hypertrophy of normal postnatal cardiac development and for an adaptive response to cardiac stress. PMID- 12782681 TI - Gliotactin, a novel marker of tricellular junctions, is necessary for septate junction development in Drosophila. AB - Septate junctions (SJs), similar to tight junctions, function as transepithelial permeability barriers. Gliotactin (Gli) is a cholinesterase-like molecule that is necessary for blood-nerve barrier integrity, and may, therefore, contribute to SJ development or function. To address this hypothesis, we analyzed Gli expression and the Gli mutant phenotype in Drosophila epithelia. In Gli mutants, localization of SJ markers neurexin-IV, discs large, and coracle are disrupted. Furthermore, SJ barrier function is lost as determined by dye permeability assays. These data suggest that Gli is necessary for SJ formation. Surprisingly, Gli distribution only colocalizes with other SJ markers at tricellular junctions, suggesting that Gli has a unique function in SJ development. Ultrastructural analysis of Gli mutants supports this notion. In contrast to other SJ mutants in which septa are missing, septa are present in Gli mutants, but the junction has an immature morphology. We propose a model, whereby Gli acts at tricellular junctions to bind, anchor, or compact SJ strands apically during SJ development. PMID- 12782682 TI - Dominant cataracts result from incongruous mixing of wild-type lens connexins. AB - Gap junctions are composed of proteins called connexins (Cx) and facilitate both ionic and biochemical modes of intercellular communication. In the lens, Cx46 and Cx50 provide the gap junctional coupling needed for homeostasis and growth. In mice, deletion of Cx46 produced severe cataracts, whereas knockout of Cx50 resulted in significantly reduced lens growth and milder cataracts. Genetic replacement of Cx50 with Cx46 by knockin rescued clarity but not growth. By mating knockin and knockout mice, we show that heterozygous replacement of Cx50 with Cx46 rescued growth but produced dominant cataracts that resulted from disruption of lens fiber morphology and crystallin precipitation. Impedance measurements revealed normal levels of ionic gap junctional coupling, whereas the passage of fluorescent dyes that mimic biochemical coupling was altered in heterozygous knockin lenses. In addition, double heterozygous knockout lenses retained normal growth and clarity, whereas knockover lenses, where native Cx46 was deleted and homozygously knocked into the Cx50 locus, displayed significantly deficient growth but maintained clarity. Together, these findings suggest that unique biochemical modes of gap junctional communication influence lens clarity and lens growth, and this biochemical coupling is modulated by the connexin composition of the gap junction channels. PMID- 12782683 TI - A genetically encoded fluorescent reporter reveals oscillatory phosphorylation by protein kinase C. AB - Signals transduced by kinases depend on the extent and duration of substrate phosphorylation. We generated genetically encoded fluorescent reporters for PKC activity that reversibly respond to stimuli activating PKC. Specifically, phosphorylation of the reporter expressed in mammalian cells causes changes in fluorescence resonance energy transfer (FRET), allowing real time imaging of phosphorylation resulting from PKC activation. Targeting of the reporter to the plasma membrane, where PKC is activated, reveals oscillatory phosphorylation in HeLa cells in response to histamine. Each oscillation in substrate phosphorylation follows a calcium oscillation with a lag of approximately 10 s. Novel FRET-based reporters for PKC translocation, phosphoinositide bisphosphate conversion to IP3, and diacylglycerol show that in HeLa cells the oscillatory phosphorylations correlate with Ca2+-controlled translocation of conventional PKC to the membrane without oscillations of PLC activity or diacylglycerol. However, in MDCK cells stimulated with ATP, PLC and diacylglycerol fluctuate together with Ca2+ and phosphorylation. Thus, specificity of PKC signaling depends on the local second messenger-controlled equilibrium between kinase and phosphatase activities to result in strict calcium-controlled temporal regulation of substrate phosphorylation. PMID- 12782684 TI - Ras recruits mitotic exit regulator Lte1 to the bud cortex in budding yeast. AB - ACdc25 family protein Lte1 (low temperature essential) is essential for mitotic exit at a lowered temperature and has been presumed to be a guanine nucleotide exchange factor (GEF) for a small GTPase Tem1, which is a key regulator of mitotic exit. We found that Lte1 physically associates with Ras2-GTP both in vivo and in vitro and that the Cdc25 homology domain (CHD) of Lte1 is essential for the interaction with Ras2. Furthermore, we found that the proper localization of Lte1 to the bud cortex is dependent on active Ras and that the overexpression of a derivative of Lte1 without the CHD suppresses defects in mitotic exit of a Deltalte1 mutant and a Deltaras1 Deltaras2 mutant. These results suggest that Lte1 is a downstream effector protein of Ras in mitotic exit and that the Ras GEF domain of Lte1 is not essential for mitotic exit but required for its localization. PMID- 12782685 TI - Nanometer targeting of microtubules to focal adhesions. AB - Although cell movement is driven by actin, polarization and directional locomotion require an intact microtubule cytoskeleton that influences polarization by modulating substrate adhesion via specific targeting interactions with adhesion complexes. The fidelity of adhesion site targeting is precise; using total internal reflection fluorescence microscopy (TIRFM), we now show microtubule ends (visualized by incorporation of GFP tubulin) are within 50 nm of the substrate when polymerizing toward the cell periphery, but not when shrinking from it. Multiple microtubules sometimes followed similar tracks, suggesting guidance along a common cytoskeletal element. Use of TIRFM with GFP- or DsRed zyxin in combination with either GFP-tubulin or GFP-CLIP-170 further revealed that the polymerizing microtubule plus ends that tracked close to the dorsal surface consistently targeted substrate adhesion complexes. This supports a central role for the microtubule tip complex in the guidance of microtubules into adhesion foci, and provides evidence for an intimate cross-talk between microtubule tips and substrate adhesions in the range of molecular dimensions. PMID- 12782687 TI - A promising therapeutic approach to spinal cord repair. PMID- 12782686 TI - Neuroglian, Gliotactin, and the Na+/K+ ATPase are essential for septate junction function in Drosophila. AB - One essential function of epithelia is to form a barrier between the apical and basolateral surfaces of the epithelium. In vertebrate epithelia, the tight junction is the primary barrier to paracellular flow across epithelia, whereas in invertebrate epithelia, the septate junction (SJ) provides this function. In this study, we identify new proteins that are required for a functional paracellular barrier in Drosophila. In addition to the previously known components Coracle (COR) and Neurexin (NRX), we show that four other proteins, Gliotactin, Neuroglian (NRG), and both the alpha and beta subunits of the Na+/K+ ATPase, are required for formation of the paracellular barrier. In contrast to previous reports, we demonstrate that the Na pump is not localized basolaterally in epithelial cells, but instead is concentrated at the SJ. Data from immunoprecipitation and somatic mosaic studies suggest that COR, NRX, NRG, and the Na+/K+ ATPase form an interdependent complex. Furthermore, the observation that NRG, a Drosophila homologue of vertebrate neurofascin, is an SJ component is consistent with the notion that the invertebrate SJ is homologous to the vertebrate paranodal SJ. These findings have implications not only for invertebrate epithelia and barrier functions, but also for understanding of neuron-glial interactions in the mammalian nervous system. PMID- 12782688 TI - The challenge of tuberculosis. PMID- 12782689 TI - An overview of the eye in diabetes. AB - The three papers in this symposium are based on presentations to an RSM meeting on the Diabetic Eye, held on 9 April 2003. The matter is particularly topical because the National Service Framework for Diabetes calls for a high-quality retinal screening programme. After a review of the various ophthalmic conditions likely to be encountered in diabetic patients (A Negi, S A Vernon) we proceed to the most important, diabetic retinopathy, with a discussion of screening methods (D M Squirrell, J F Talbot) and an account of laser treatments (J G F Dowler). Colour versions of the clinical photographs are available online [www.jrsm.org]. Publication was coordinated by Professor Susan Lightman, of Moorfields Eye Hospital, London, UK. PMID- 12782691 TI - Laser management of diabetic retinopathy. PMID- 12782690 TI - Screening for diabetic retinopathy. PMID- 12782692 TI - Breast cancer and the pill. PMID- 12782693 TI - Brain tuned to music. PMID- 12782694 TI - Myeloma and a mass in the heart. PMID- 12782695 TI - An Afghan child with deep vein thrombosis. PMID- 12782696 TI - Mycotic aortic aneurysm with protective coarctation. PMID- 12782697 TI - Periorbital cellulitis with breast cancer. PMID- 12782698 TI - Retroperitoneal haemorrhage during warfarin therapy. PMID- 12782699 TI - Sylvia Plath and the depression continuum. PMID- 12782700 TI - The master of myomectomy. PMID- 12782701 TI - A tale of two ears. PMID- 12782702 TI - Standard care. PMID- 12782703 TI - Hyperthyroidism with low thyroid hormone. PMID- 12782704 TI - What's important in the doctor-patient relationship? PMID- 12782705 TI - Cocaine by internal mail. PMID- 12782706 TI - Consent to treatment and mental health. PMID- 12782707 TI - Hyperthyroidism with low thyroid hormone. PMID- 12782708 TI - Exploiting the redundancy in the immune system: vaccines can mediate protection by eliciting 'unnatural' immunity. PMID- 12782709 TI - Vaccine immunity to pathogenic fungi overcomes the requirement for CD4 help in exogenous antigen presentation to CD8+ T cells: implications for vaccine development in immune-deficient hosts. AB - Systemic fungal infections with primary and opportunistic pathogens have become increasingly common and represent a growing health menace in patients with AIDS and other immune deficiencies. T lymphocyte immunity, in particular the CD4+ Th 1 cells, is considered the main defense against these pathogens, and their absence is associated with increased susceptibility. It would seem illogical then to propose vaccinating these vulnerable patients against fungal infections. We report here that CD4+ T cells are dispensable for vaccine-induced resistance against experimental fungal pulmonary infections with two agents, Blastomyces dermatitidis an extracellular pathogen, and Histoplasma capsulatum a facultative intracellular pathogen. In the absence of T helper cells, exogenous fungal antigens activated memory CD8+ cells in a major histocompatibility complex class I-restricted manner and CD8+ T cell-derived cytokines tumor necrosis factor alpha, interferon gamma, and granulocyte/macrophage colony-stimulating factor mediated durable vaccine immunity. CD8+ T cells could also rely on alternate mechanisms for robust vaccine immunity, in the absence of some of these factors. Our results demonstrate an unexpected plasticity of immunity in compromised hosts at both the cellular and molecular level and point to the feasibility of developing vaccines against invasive fungal infections in patients with severe immune deficiencies, including those with few or no CD4+ T cells. PMID- 12782710 TI - Human cytomegalovirus glycoprotein UL16 causes intracellular sequestration of NKG2D ligands, protecting against natural killer cell cytotoxicity. AB - The activating receptor, NKG2D, is expressed on a variety of immune effector cells and recognizes divergent families of major histocompatibility complex (MHC) class I-related ligands, including the MIC and ULBP proteins. Infection, stress, or transformation can induce NKG2D ligand expression, resulting in effector cell activation and killing of the ligand-expressing target cell. The human cytomegalovirus (HCMV) membrane glycoprotein, UL16, binds to three of the five known ligands for human NKG2D. UL16 is retained in the endoplasmic reticulum and cis-Golgi apparatus of cells and causes MICB to be similarly retained and stabilized within cells. Coexpression of UL16 markedly reduces cell surface levels of MICB, ULBP1, and ULBP2, and decreases susceptibility to natural killer cell-mediated cytotoxicity. Domain swapping experiments demonstrate that the transmembrane and cytoplasmic domains of UL16 are important for intracellular retention of UL16, whereas the ectodomain of UL16 participates in down-regulation of NKG2D ligands. The intracellular sequestration of NKG2D ligands by UL16 represents a novel HCMV immune evasion mechanism to add to the well-documented viral strategies directed against antigen presentation by classical MHC molecules. PMID- 12782711 TI - Orphan nuclear receptor Nur77 is involved in caspase-independent macrophage cell death. AB - Activation-induced cell death in macrophages has been observed, but the mechanism remains largely unknown. Activation-induced cell death in macrophages can be independent from caspases, and the death of activated macrophages can even be triggered by the pan-caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone (zVAD). Here, we show that this type of macrophage death can occur in the septic mouse model and that toll-like receptor (TLR)-2 or TLR4 signaling is required in this process. We conclude that Nur77 is involved in the macrophage death because Nur77 expression correlates with cell death, and cell death is reduced significantly in Nur77-deficient macrophages. The extracellular signal regulated kinase pathway, which is downstream of TLR2 or TLR4, and myocyte specific enhancer binding factor 2 (MEF2) transcription factor activity, which is up-regulated by zVAD, are required for Nur77 induction and macrophage death. Reporter gene analysis suggests that Nap, Ets, Rce, and Sp1 sites in the Nur77 promoter are regulated by TLR4 signaling and that MEF2 sites in the Nur77 promoter are regulated by zVAD treatment. MEF2 transcription factors are constitutively expressed and degraded in macrophages, and zVAD increases MEF2 transcription factor activity by preventing the proteolytic cleavage and degradation of MEF2 proteins. This paper delineates the dual signaling pathways that are required for Nur77 induction in macrophages and demonstrates a role of Nur77 in caspase-independent cell death. PMID- 12782712 TI - Preferential signaling and induction of allergy-promoting lymphokines upon weak stimulation of the high affinity IgE receptor on mast cells. AB - Mast cell degranulation and de novo cytokine production is a consequence of antigen-aggregation of the immunoglobulin E (IgE)-occupied high affinity receptor for IgE (Fc epsilon RI). Herein, we report that lymphokines that promote allergic inflammation, like MCP-1, were potently induced at low antigen (Ag) concentrations or at low receptor occupancy with IgE whereas some that down regulate this response, like interleukin (IL)-10, required high receptor occupancy. Weak stimulation of mast cells caused minimal degranulation whereas a half-maximal secretory response was observed for chemokines and, with the exception of TNF-alpha, a weaker cytokine secretory response was observed. The medium from weakly stimulated mast cells elicited a monocyte/macrophage chemotactic response similar to that observed at high receptor occupancy. Weak stimulation also favored the phosphorylation of Gab2 and p38MAPK, while LAT and ERK2 phosphorylation was induced by a stronger stimulus. Gab2-deficient mast cells were severely impaired in chemokine mRNA induction whereas LAT-deficient mast cells showed a more pronounced defect in cytokines. These findings demonstrate that perturbation of small numbers of IgE receptors on mast cells favors certain signals that contribute to a lymphokine response that can mediate allergic inflammation. PMID- 12782713 TI - MIF signal transduction initiated by binding to CD74. AB - Macrophage migration inhibitory factor (MIF) accounts for one of the first cytokine activities to have been described, and it has emerged recently to be an important regulator of innate and adaptive immunity. MIF is an upstream activator of monocytes/macrophages, and it is centrally involved in the pathogenesis of septic shock, arthritis, and other inflammatory conditions. The protein is encoded by a unique but highly conserved gene, and X-ray crystallography studies have shown MIF to define a new protein fold and structural superfamily. Although recent work has begun to illuminate the signal transduction pathways activated by MIF, the nature of its membrane receptor has not been known. Using expression cloning and functional analysis, we report herein that CD74, a Type II transmembrane protein, is a high-affinity binding protein for MIF. MIF binds to the extracellular domain of CD74, and CD74 is required for MIF-induced activation of the extracellular signal-regulated kinase-1/2 MAP kinase cascade, cell proliferation, and PGE2 production. A recombinant, soluble form of CD74 binds MIF with a dissociation constant of approximately 9 x 10-9 Kd, as defined by surface plasmon resonance (BIAcore analysis), and soluble CD74 inhibits MIF-mediated extracellular signal-regulated kinase activation in defined cell systems. These data provide a molecular basis for MIF's interaction with target cells and identify it as a natural ligand for CD74, which has been implicated previously in signaling and accessory functions for immune cell activation. PMID- 12782714 TI - Classification and prediction of survival in patients with the leukemic phase of cutaneous T cell lymphoma. AB - We have used cDNA arrays to investigate gene expression patterns in peripheral blood mononuclear cells from patients with leukemic forms of cutaneous T cell lymphoma, primarily Sezary syndrome (SS). When expression data for patients with high blood tumor burden (Sezary cells >60% of the lymphocytes) and healthy controls are compared by Student's t test, at P < 0.01, we find 385 genes to be differentially expressed. Highly overexpressed genes include Th2 cells-specific transcription factors Gata-3 and Jun B, as well as integrin beta1, proteoglycan 2, the RhoB oncogene, and dual specificity phosphatase 1. Highly underexpressed genes include CD26, Stat-4, and the IL-1 receptors. Message for plastin-T, not normally expressed in lymphoid tissue, is detected only in patient samples and may provide a new marker for diagnosis. Using penalized discriminant analysis, we have identified a panel of eight genes that can distinguish SS in patients with as few as 5% circulating tumor cells. This suggests that, even in early disease, Sezary cells produce chemokines and cytokines that induce an expression profile in the peripheral blood distinctive to SS. Finally, we show that using 10 genes, we can identify a class of patients who will succumb within six months of sampling regardless of their tumor burden. PMID- 12782715 TI - Protein kinase C theta affects Ca2+ mobilization and NFAT cell activation in primary mouse T cells. AB - Protein kinase C (PKC)theta is an established component of the immunological synapse and has been implicated in the control of AP-1 and NF-kappaB. To study the physiological function of PKCtheta, we used gene targeting to generate a PKCtheta null allele in mice. Consistently, interleukin 2 production and T cell proliferative responses were strongly reduced in PKCtheta-deficient T cells. Surprisingly, however, we demonstrate that after CD3/CD28 engagement, deficiency of PKCtheta primarily abrogates NFAT transactivation. In contrast, NF-kappaB activation was only partially reduced. This NFAT transactivation defect appears to be secondary to reduced inositol 1,4,5-trisphosphate generation and intracellular Ca2+ mobilization. Our finding suggests that PKCtheta plays a critical and nonredundant role in T cell receptor-induced NFAT activation. PMID- 12782716 TI - An alternatively spliced variant of CXCR3 mediates the inhibition of endothelial cell growth induced by IP-10, Mig, and I-TAC, and acts as functional receptor for platelet factor 4. AB - The chemokines CXCL9/Mig, CXCL10/IP-10, and CXCL11/I-TAC regulate lymphocyte chemotaxis, mediate vascular pericyte proliferation, and act as angiostatic agents, thus inhibiting tumor growth. These multiple activities are apparently mediated by a unique G protein-coupled receptor, termed CXCR3. The chemokine CXCL4/PF4 shares several activities with CXCL9, CXCL10, and CXCL11, including a powerful angiostatic effect, but its specific receptor is still unknown. Here, we describe a distinct, previously unrecognized receptor named CXCR3-B, derived from an alternative splicing of the CXCR3 gene that mediates the angiostatic activity of CXCR3 ligands and also acts as functional receptor for CXCL4. Human microvascular endothelial cell line-1 (HMEC-1), transfected with either the known CXCR3 (renamed CXCR3-A) or CXCR3-B, bound CXCL9, CXCL10, and CXCL11, whereas CXCL4 showed high affinity only for CXCR3-B. Overexpression of CXCR3-A induced an increase of survival, whereas overexpression of CXCR3-B dramatically reduced DNA synthesis and up-regulated apoptotic HMEC-1 death through activation of distinct signal transduction pathways. Remarkably, primary cultures of human microvascular endothelial cells, whose growth is inhibited by CXCL9, CXCL10, CXCL11, and CXCL4, expressed CXCR3-B, but not CXCR3-A. Finally, monoclonal antibodies raised to selectively recognize CXCR3-B reacted with endothelial cells from neoplastic tissues, providing evidence that CXCR3-B is also expressed in vivo and may account for the angiostatic effects of CXC chemokines. PMID- 12782717 TI - The lectin-like receptor KLRE1 inhibits natural killer cell cytotoxicity. AB - We report the cloning and functional characterization in the mouse and the rat of a novel natural killer (NK) cell receptor termed KLRE1. The receptor is a type II transmembrane protein with a COOH-terminal lectin-like domain, and constitutes a novel KLR family. Rat Klre1 was mapped to the NK gene complex. By Northern blot and flow cytometry using newly generated monoclonal antibodies, KLRE1 was shown to be expressed by NK cells and a subpopulation of CD3+ cells, with pronounced interstrain variation. Western blot analysis indicated that KLRE1 can be expressed on the NK cell surface as a disulphide-linked dimer. The predicted proteins do not contain immunoreceptor tyrosine-based inhibitory motifs (ITIMs) or a positively charged amino acid in the transmembrane domain. However, in a redirected lysis assay, the presence of whole IgG, but not of F(ab')2 fragments of a monoclonal anti-KLRE1 antibody inhibited lysis of Fc-receptor bearing tumor target cells. Moreover, the tyrosine phosphatase SHP-1 was coimmunoprecipitated with KLRE1 from pervanadate-treated interleukin 2-activated NK cells. Together, our results indicate that KLRE1 may form a functional heterodimer with an as yet unidentified ITIM-bearing partner that recruits SHP-1 to generate an inhibitory receptor complex. PMID- 12782718 TI - Marrow stem cells shift gene expression and engraftment phenotype with cell cycle transit. AB - We studied the genetic and engraftment phenotype of highly purified murine hematopoietic stem cells (lineage negative, rhodamine-low, Hoechst-low) through cytokine-stimulated cell cycle. Cells were cultured in interleukin (IL)-3, IL-6, IL-11, and steel factor for 0 to 48 h and tested for engraftment capacity in a lethally irradiated murine competitive transplant model. Engraftment showed major fluctuations with nadirs at 36 and 48 h of culture and recovery during the next G1. Gene expression of quiescent (0 h) or cycling (48 h) stem cells was compared with lineage positive cells by 3' end PCR differential display analysis. Individual PCR bands were quantified using a 0 to 9 scale and results were visually compared using color-coded matrices. We defined a set of 637 transcripts expressed in stem cells and not expressed in lineage positive cells. Gene expression analyzed at 0 and 48 h showed a major shift from "stem cell genes" being highly expressed at 0 h and turned off at 48 h, while "cell division" genes were turned on at 48 h. These observations suggest stem cell gene expression shifts through cell cycle in relation to cell cycle related alterations of stem cell phenotype. The engraftment defect is related to a major phenotypic change of the stem cell. PMID- 12782719 TI - Inhibition of interleukin 10 signaling after Fc receptor ligation and during rheumatoid arthritis. AB - Interleukin-10 (IL-10) is a potent deactivator of myeloid cells that limits the intensity and duration of immune and inflammatory responses. The activity of IL 10 can be suppressed during inflammation, infection, or after allogeneic tissue transplantation. We investigated whether inflammatory factors suppress IL-10 activity at the level of signal transduction. Out of many factors tested, only ligation of Fc receptors by immune complexes inhibited IL-10 activation of the Jak-Stat signaling pathway. IL-10 signaling was suppressed in rheumatoid arthritis joint macrophages that are exposed to immune complexes in vivo. Activation of macrophages with interferon-gamma was required for Fc receptor mediated suppression of IL-10 signaling, which resulted in diminished activation of IL-10-inducible genes and reversal of IL-10-dependent suppression of cytokine production. The mechanism of inhibition involved decreased cell surface IL-10 receptor expression and Jak1 activation and was dependent on protein kinase C delta. These results establish that IL-10 signaling is regulated during inflammation and identify Fc receptors and interferon-gamma as important regulators of IL-10 activity. Generation of macrophages refractory to IL-10 can contribute to pathogenesis of inflammatory and infectious diseases characterized by production of interferon-gamma and immune complexes. PMID- 12782720 TI - Accumulation of tissue factor into developing thrombi in vivo is dependent upon microparticle P-selectin glycoprotein ligand 1 and platelet P-selectin. AB - Using a laser-induced endothelial injury model, we examined thrombus formation in the microcirculation of wild-type and genetically altered mice by real-time in vivo microscopy to analyze this complex physiologic process in a system that includes the vessel wall, the presence of flowing blood, and the absence of anticoagulants. We observe P-selectin expression, tissue factor accumulation, and fibrin generation after platelet localization in the developing thrombus in arterioles of wild-type mice. However, mice lacking P-selectin glycoprotein ligand 1 (PSGL-1) or P-selectin, or wild-type mice infused with blocking P selectin antibodies, developed platelet thrombi containing minimal tissue factor and fibrin. To explore the delivery of tissue factor into a developing thrombus, we identified monocyte-derived microparticles in human platelet-poor plasma that express tissue factor, PSGL-1, and CD14. Fluorescently labeled mouse microparticles infused into a recipient mouse localized within the developing thrombus, indicating that one pathway for the initiation of blood coagulation in vivo involves the accumulation of tissue factor- and PSGL-1-containing microparticles in the platelet thrombus expressing P-selectin. These monocyte derived microparticles bind to activated platelets in an interaction mediated by platelet P-selectin and microparticle PSGL-1. We propose that PSGL-1 plays a role in blood coagulation in addition to its known role in leukocyte trafficking. PMID- 12782722 TI - Role of nicotianamine in the intracellular delivery of metals and plant reproductive development. AB - Nicotianamine (NA), a chelator of metals, is ubiquitously present in higher plants. Nicotianamine aminotransferase (NAAT) catalyzes the amino group transfer of NA in the biosynthetic pathway of phytosiderophores and is essential for iron acquisition in graminaceous plants. The gene that encodes NAAT from barley was introduced into the nongraminaceous plant tobacco, which produces NA but not phytosiderophores. Transgenic tobacco plants (naat tobacco) that constitutively expressed the NAAT gene had young leaves with interveinal chlorosis and flowers that were abnormally shaped and sterile. Endogenous NA was consumed as a result of NAAT overproduction in naat tobacco. The resulting NA shortage caused disorders in internal metal transport, leading to these abnormal phenotypes. In addition to its role in long-distance metal transport, NA may be involved in the regulation of metal transfer within the cells. These results suggest that a shortage of NA impaired the functions of metal-requiring proteins, including transcription factors. PMID- 12782724 TI - Regulatory elements of the floral homeotic gene AGAMOUS identified by phylogenetic footprinting and shadowing. AB - In Arabidopsis thaliana, cis-regulatory sequences of the floral homeotic gene AGAMOUS (AG) are located in the second intron. This 3-kb intron contains binding sites for two direct activators of AG, LEAFY (LFY) and WUSCHEL (WUS), along with other putative regulatory elements. We have used phylogenetic footprinting and the related technique of phylogenetic shadowing to identify putative cis regulatory elements in this intron. Among 29 Brassicaceae species, several other motifs, but not the LFY and WUS binding sites identified previously, are largely invariant. Using reporter gene analyses, we tested six of these motifs and found that they are all functionally important for the activity of AG regulatory sequences in A. thaliana. Although there is little obvious sequence similarity outside the Brassicaceae, the intron from cucumber AG has at least partial activity in A. thaliana. Our studies underscore the value of the comparative approach as a tool that complements gene-by-gene promoter dissection but also demonstrate that sequence-based studies alone are insufficient for a complete identification of cis-regulatory sites. PMID- 12782723 TI - The Arabidopsis MALE MEIOCYTE DEATH1 gene encodes a PHD-finger protein that is required for male meiosis. AB - In plants, reproductive development requires normal meiosis, which involved several highly coordinated events. Such meiotic events are regulated in a number of ways in yeast and animal systems, including transcriptional and checkpoint control mechanisms. Although a number of mutations that affect different aspects of meiosis have been characterized in plants, very little is known about the regulation of plant meiosis at the molecular level. In particular, no meiosis specific transcriptional regulators have been identified in plants, and checkpoint control has not been observed during plant meiosis. We report here the isolation and characterization of a new Arabidopsis male-sterile mutant that exhibits meiotic defects. Meiocytes from mutant plants appeared normal up to diakinesis, when they exhibited signs of apoptosis, including defects in chromosome behavior, cytoplasmic shrinkage, and chromatin fragmentation, followed by cell death before cytokinesis. Therefore, the mutant was named male meiocyte death1 (mmd1). The MMD1 gene was cloned using Dissociation transposon tagging and encodes a plant homeo domain domain-containing protein. MMD1 is expressed preferentially during male meiosis. Our results suggest that MMD1 may be involved in the regulation of gene expression during meiosis and that the mmd1 mutation triggers cell death in male meiocytes. PMID- 12782725 TI - Arabidopsis SGT1b is required for SCF(TIR1)-mediated auxin response. AB - The SCF(TIR1) complex is a central regulator of the auxin response pathway in Arabidopsis. This complex functions as a ubiquitin protein ligase that targets members of the auxin/indoleacetic acid (Aux/IAA) family of transcriptional regulators for ubiquitin-mediated degradation in response to auxin. In an attempt to identify additional factors required for SCF(TIR1) activity, we conducted a genetic screen to isolate enhancers of the auxin response defect conferred by the tir1-1 mutation. Here, we report the identification and characterization of the eta3 mutant. The eta3 mutation interacts synergistically with tir1-1 to strongly enhance all aspects of the tir1 mutant phenotype, including auxin inhibition of root growth, lateral root development, hypocotyl elongation at high temperature, and apical dominance. We isolated the ETA3 gene using a map-based cloning strategy and determined that ETA3 encodes SGT1b. SGT1b was identified recently as a factor involved in plant disease resistance signaling, and SGT1 from barley and tobacco extracts was shown to interact with SCF ubiquitin ligases. We conclude that ETA3/SGT1b is required for the SCF(TIR1)-mediated degradation of Aux/IAA proteins. PMID- 12782726 TI - Functional analysis of beta- and epsilon-ring carotenoid hydroxylases in Arabidopsis. AB - Lutein and zeaxanthin are dihydroxy xanthophylls that are produced from their corresponding carotene precursors by the action of beta- and epsilon -ring carotenoid hydroxylases. Two genes that encode beta-ring hydroxylases (beta hydroxylases 1 and 2) have been identified in the Arabidopsis genome and are highly active toward beta-rings but only weakly active toward epsilon -rings. A third distinct activity required for epsilon -ring hydroxylation has been defined by mutation of the LUTEIN1 (LUT1) locus, but LUT1 has not yet been cloned. To address the individual and overlapping functions of the three Arabidopsis carotenoid hydroxylase activities in vivo, T-DNA knockout mutants corresponding to beta-hydroxylases 1 and 2 (b1 and b2, respectively) were isolated and all possible hydroxylase mutant combinations were generated. beta-Hydroxylase single mutants do not exhibit obvious growth defects and have limited impact on carotenoid composition relative to the wild type, suggesting that the encoded proteins have a significant degree of functional redundancy in vivo. Surprisingly, the b1 b2 double mutant, which lacks both known beta-hydroxylase enzymes, still contains significant levels of beta-carotene-derived xanthophylls, suggesting that additional beta-ring hydroxylation activity exists in vivo. The phenotype of double and triple hydroxylase mutants indicates that at least a portion of this activity resides in the LUT1 gene product. Despite the severe reduction of beta-carotene-derived xanthophylls (up to 90% in the lut1 b1 b2 triple mutant), the double and triple hydroxylase mutants still contain at least 50% of the wild-type amount of hydroxylated beta-rings. This finding suggests that it is the presence of minimal amounts of hydroxylated beta-rings, rather than minimal amounts of specific beta-carotene-derived xanthophylls, that are essential for light-harvesting complex II assembly and function in vivo. The carotenoid profiles in wild-type seeds and the effect of single and multiple hydroxylase mutations are distinct from those in photosynthetic tissues, indicating that the activities of each gene product differ in the two tissues. Overall, the hydroxylase mutants provide insight into the unexpected overlapping activity of carotenoid hydroxylases in vivo. PMID- 12782727 TI - PAA1, a P-type ATPase of Arabidopsis, functions in copper transport in chloroplasts. AB - Copper (Cu) is an essential trace element with important roles as a cofactor in many plant functions, including photosynthesis. However, free Cu ions can cause toxicity, necessitating precise Cu delivery systems. Relatively little is known about Cu transport in plant cells, and no components of the Cu transport machinery in chloroplasts have been identified previously. Cu transport into chloroplasts provides the cofactor for the stromal enzyme copper/zinc superoxide dismutase (Cu/ZnSOD) and for the thylakoid lumen protein plastocyanin, which functions in photosynthetic electron transport from the cytochrome b(6)f complex to photosystem I. Here, we characterized six Arabidopsis mutants that are defective in the PAA1 gene, which encodes a member of the metal-transporting P type ATPase family with a functional N-terminal chloroplast transit peptide. paa1 mutants exhibited a high-chlorophyll-fluorescence phenotype as a result of an impairment of photosynthetic electron transport that could be ascribed to decreased levels of holoplastocyanin. The paa1-1 mutant had a lower chloroplast Cu content, despite having wild-type levels in leaves. The electron transport defect of paa1 mutants was evident on medium containing <1 micro M Cu, but it was suppressed by the addition of 10 micro M Cu. Chloroplastic Cu/ZnSOD activity also was reduced in paa1 mutants, suggesting that PAA1 mediates Cu transfer across the plastid envelope. Thus, PAA1 is a critical component of a Cu transport system in chloroplasts responsible for cofactor delivery to plastocyanin and Cu/ZnSOD. PMID- 12782728 TI - Small ubiquitin-like modifier modulates abscisic acid signaling in Arabidopsis. AB - Post-translational modification of proteins by small polypeptides, such as ubiquitin, has emerged as a common and important mechanism for regulating protein function. Small ubiquitin-like modifier (SUMO) is a small protein that is structurally related to but functionally different from ubiquitin. We report the identification and functional analysis of AtSUMO1, AtSUMO2, and AtSCE1a as components of the SUMO conjugation (sumoylation) pathway in Arabidopsis. In yeast two hybrid assays, AtSUMO1/2 interacts specifically with a SUMO-conjugating enzyme but not with a ubiquitin-conjugating enzyme. AtSCE1a, the Arabidopsis SUMO conjugating enzyme ortholog, conjugates SUMO to RanGAP in vitro. AtSUMO1/2 and AtSCE1a colocalize at the nucleus, and AtSUMO1/2 are conjugated to endogenous SUMO targets in vivo. Analysis of transgenic plants showed that overexpression of AtSUMO1/2 does not have any obvious effect in general plant development, but increased sumoylation levels attenuate abscisic acid (ABA)-mediated growth inhibition and amplify the induction of ABA- and stress-responsive genes such as RD29A. Reduction of AtSCE1a expression levels accentuates ABA-mediated growth inhibition. Our results suggest a role for SUMO in the modulation of the ABA signal transduction pathway. PMID- 12782729 TI - Inhibition of cell growth and shoot development by a specific nucleotide sequence in a noncoding viroid RNA. AB - Viroids are small noncoding and infectious RNAs that replicate autonomously and move systemically throughout an infected plant. The RNAs of the family Pospiviroidae contain a central conserved region (CCR) that has long been thought to be involved in replication. Here, we report that the CCR of Potato spindle tuber viroid (PSTVd) also plays a role in pathogenicity. A U257A change in the CCR converted the intermediate strain PSTVd(Int) to a lethal strain that caused severe growth stunting and premature death of infected plants. PSTVd with nucleotide U257 changed to C or G did not cause such symptoms. The pathogenic effect of the U257A substitution was abolished by a C259U substitution in the same RNA. Analyses of the pathogenic effects of the U257A substitution in three other PSTVd variants established A257 as a new pathogenicity determinant that functions independently and synergistically with the classic pathogenicity domain. The U257A substitution did not alter PSTVd secondary structure, replication levels, or tissue tropism. The stunted growth of PSTVd(Int)U257A infected tomato plants resulted from restricted cell expansion but not cell division or differentiation. This was correlated positively with the downregulated expression of an expansin gene, LeExp2. Our results demonstrate that specific nucleotides in a noncoding, pathogenic RNA have a profound effect in altering distinct cellular responses, which then lead to well-defined alterations in plant growth and developmental patterns. The feasibility of correlating viroid RNA sequence/structure with the altered expression of specific host genes, cellular processes, and developmental patterns makes viroid infection a valuable system in which to investigate host factors for symptom expression and perhaps also to characterize the mechanisms of RNA regulation of gene expression in plants. PMID- 12782730 TI - PmSUC3: characterization of a SUT2/SUC3-type sucrose transporter from Plantago major. AB - Higher plants possess medium-sized gene families that encode plasma membrane localized sucrose transporters. For several plant species, it has been shown that at least one of these genes (e.g., AtSUC3 in Arabidopsis and LeSUT2 in tomato) differs from all other family members in several features, such as the length of the open reading frame, the number of introns, and the codon usage bias. For these reasons, and because two of these proteins did not rescue a yeast mutant defective in sucrose utilization, it had been speculated that this subgroup of transporters might have sensor functions. Here, we describe the detailed functional characterization and cellular localization of PmSUC3, the orthologous transporter from the Plantago major transporter family. The PmSUC3 protein is localized in the sieve elements of the Plantago phloem and mediates the energy dependent transport of sucrose and maltose. In contrast to the situation in solanaceous plants, PmSUC3 is not colocalized with PmSUC2, the source-specific, phloem-loading sucrose transporter of Plantago. Moreover, PmSUC3 also was identified in sieve elements of sink leaves and in several nonphloem cells and tissues. Arguments for and against a potential sensor function for this type of sucrose transporter are presented, and the role of this type of transporter in the regulation of sucrose fluxes is discussed. PMID- 12782731 TI - Regulation of expansin gene expression affects growth and development in transgenic rice plants. AB - To investigate the in vivo functions of expansins, we generated transgenic rice plants that express sense and antisense constructs of the expansin gene OsEXP4. In adult plants with constitutive OsEXP4 expression, 12% of overexpressors were taller and 88% were shorter than the average control plants, and most overexpressors developed at least two additional leaves. Antisense plants were shorter and flowered earlier than the average control plants. In transgenic plants with inducible OsEXP4 expression, we observed a close correlation between OsEXP4 protein levels and seedling growth. Coleoptile and mesocotyl length increased by up to 31 and 97%, respectively, in overexpressors, whereas in antisense seedlings, they decreased by up to 28 and 43%, respectively. The change in seedling growth resulted from corresponding changes in cell size, which in turn appeared to be a function of altered cell wall extensibility. Our results support the hypothesis that expansins are involved in enhancing growth by mediating cell wall loosening. PMID- 12782732 TI - 14-3-3 proteins are constituents of the insoluble glycoprotein framework of the chlamydomonas cell wall. AB - The cell wall of the unicellular green alga Chlamydomonas reinhardtii consists predominantly of Hyp-rich glycoproteins, which also occur in the extracellular matrix of multicellular green algae and higher plants. In addition to the Hyp rich polypeptides, the insoluble glycoprotein framework of the Chlamydomonas cell wall contains minor amounts of 14-3-3 proteins, as revealed by immunochemical studies and mass spectroscopic analysis of tryptic peptides. Polypeptides immunologically related to the 14-3-3 proteins also were found in the culture medium of Chlamydomonas. The levels of two of these 14-3-3-related polypeptides were decreased in the culture medium of the wall-deficient mutant cw-15. These findings indicate that 14-3-3 proteins are involved in the cross-linking of Hyp rich glycoproteins in the Chlamydomonas cell wall. PMID- 12782733 TI - Mutation or drug-dependent microtubule disruption causes radial swelling without altering parallel cellulose microfibril deposition in Arabidopsis root cells. AB - As critical determinants of growth anisotropy in plants, cortical microtubules are thought to constrain the movement of cellulose synthase complexes and thus align newly deposited cellulose microfibrils. We tested this cellulose synthase constraint model using the temperature-sensitive mor1-1 mutant of Arabidopsis. Contrary to predictions, the disruption of cortical microtubules in mor1-1 root epidermal cells led to left-handed root twisting and radial swelling but did not alter the transverse orientation of cellulose microfibrils. We also found that drug-dependent disassembly or hyperstabilization of cortical microtubules did not alter the parallel order of cellulose microfibrils. By measuring cellulose content in mor1-1 seedlings, we verified that cellulose synthesis is not reduced at the restrictive temperature. The independence of cortical microtubule organization and cellulose microfibril alignment was supported by the observation that double mutants of mor1-1 and rsw1-1, the cellulose-deficient mutant with misaligned microfibrils, had additive phenotypes. Our results suggest that cortical microtubules regulate growth anisotropy by some mechanism other than cellulose microfibril alignment or synthesis. PMID- 12782734 TI - The basis of natural and artificial postzygotic hybridization barriers in Arabidopsis species. AB - The success or failure of interspecific crosses is vital to evolution and to agriculture, but much remains to be learned about the nature of hybridization barriers. Several mechanisms have been proposed to explain postzygotic barriers, including negative interactions between diverged sequences, global genome rearrangements, and widespread epigenetic reprogramming. Another explanation is imbalance of paternally and maternally imprinted genes in the endosperm. Interspecific crosses between diploid Arabidopsis thaliana as the seed parent and tetraploid Arabidopsis arenosa as the pollen parent produced seeds that aborted with the same paternal excess endosperm phenotype seen in crosses between diploid and hexaploid A. thaliana. Doubling maternal ploidy restored seed viability and normal endosperm morphology. However, substituting a hypomethylated tetraploid A. thaliana seed parent reestablished the hybridization barrier by causing seed abortion and a lethal paternal excess phenotype. We conclude from these findings that the dominant cause of seed abortion in the diploid A. thaliana x tetraploid A. arenosa cross is parental genomic imbalance. Our results also demonstrate that manipulation of DNA methylation can be sufficient to erect hybridization barriers, offering a potential mechanism for speciation and a means of controlling gene flow between species. PMID- 12782735 TI - Cytochrome f translation in Chlamydomonas chloroplast is autoregulated by its carboxyl-terminal domain. AB - The rate of synthesis of cytochrome f is decreased approximately 10-fold when it does not assemble with the other subunits of the cytochrome b(6)f complex in Chlamydomonas reinhardtii chloroplasts. This assembly-mediated regulation of cytochrome f synthesis corresponds to a regulation of petA mRNA initiation of translation. Here, we demonstrate that cytochrome f translation is autoregulated by its C-terminal domain. Five cytochrome f residues conserved throughout all chloroplast genomes-residue Gln-297 in the transmembrane helix and a cluster of four amino acids, Lys-Gln-Phe-Glu, at positions 305 to 308, in the stromal extension-participate in the formation of a translation repressor motif. By contrast, positively charged residues in the stromal extension have little influence on the autoregulation process. These results do not favor a direct interaction between the repressor motif and the petA 5' untranslated region but suggest the participation of a membrane-bound ternary effector. PMID- 12782736 TI - Low glutelin content1: a dominant mutation that suppresses the glutelin multigene family via RNA silencing in rice. AB - Low glutelin content1 (Lgc1) is a dominant mutation that reduces glutelin content in rice grains. Glutelin is a major seed storage protein encoded by a multigene family. RNA gel blot and reverse transcriptase-mediated PCR analyses revealed that Lgc1 acts at the mRNA level in a similarity-dependent manner. In Lgc1 homozygotes, there is a 3.5-kb deletion between two highly similar glutelin genes that forms a tail-to-tail inverted repeat, which might produce a double-stranded RNA molecule, a potent inducer of RNA silencing. The hypothesis that Lgc1 suppresses glutelin expression via RNA silencing is supported by transgenic analysis using this Lgc1 candidate region, by reporter gene analysis, and by the detection of small interfering RNAs. In this context, Lgc1 provides an interesting example of RNA silencing occurring among genes that exhibit various levels of similarity to an RNA-silencing-inducing gene. Possible mechanisms for gene silencing of the glutelin multigene family by Lgc1 are discussed. PMID- 12782737 TI - The transcriptional enhancer of the pea plastocyanin gene associates with the nuclear matrix and regulates gene expression through histone acetylation. AB - The influence of the transcriptional enhancer of the pea plastocyanin gene (PetE) on the acetylation of histones was examined with chromatin immunoprecipitation (ChIP) experiments using antibodies that recognize acetylated or nonacetylated histones H3 and H4. In transgenic tobacco plants containing the pea PetE promoter fused to uidA, both acetylated and nonacetylated histones H3 and H4 were present on the integrated transgene. Linking the PetE enhancer to the transgene resulted in increased beta-glucuronidase activity and increased amounts of acetylated histones H3 and H4 present on the promoter, suggesting that the enhancer may increase transcription by mediating the acetylation of histones. Trichostatin A and sodium butyrate, which are potent inhibitors of histone deacetylases (HDAs), activated expression from the PetE promoter by fourfold, with a concomitant increase in the acetylation states of histones H3 and H4, as determined by ChIP, indicating that the acetylation of histones has a direct positive effect on transcription. The HDA inhibitors did not increase expression from the PetE promoter when it was linked to the enhancer, consistent with preexisting hyperacetylated histones on the transgene. Mapping of histone acetylation states along the reporter gene indicated that the histones H3 and H4 associated with the promoter and the 5' region of uidA were hyperacetylated in the presence of the PetE enhancer. The PetE enhancer bound to isolated tobacco nuclear matrices in vitro and was associated with the nuclear matrix in nuclei isolated from transgenic tobacco plants. These results suggest that the pea PetE enhancer activates transcription by associating with the nuclear matrix, mediating the acetylation of histones on the promoter and the nearby coding region and resulting in an altered chromatin structure. PMID- 12782739 TI - Fatigue at work. PMID- 12782738 TI - HCF152, an Arabidopsis RNA binding pentatricopeptide repeat protein involved in the processing of chloroplast psbB-psbT-psbH-petB-petD RNAs. AB - The psbB-psbT-psbH-petB-petD operon of higher plant chloroplasts is a heterogeneously composed transcriptional unit that undergoes complex RNA processing events until the mature oligocistronic RNAs are formed. To identify the nucleus-encoded factors required for the processing and expression of psbB psbT-psbH-petB-petD transcripts, we performed mutational analysis using Arabidopsis. The allelic nuclear mutants hcf152-1 and hcf152-2 were identified that are affected specifically in the accumulation of the plastidial cytochrome b(6)f complex. In both mutants, reduced amounts of spliced petB RNAs (encoding the cytochrome b(6) subunit) were detected, thus explaining the observed protein deficiencies. Additionally, mutant hcf152-1 is affected in the accumulation of transcripts cleaved between the genes psbH and petB. As a result of a close T-DNA insertion, the HCF152 gene was cloned and its identity confirmed by complementation of homozygous mutant plants. HCF152 encodes a pentatricopeptide repeat (PPR) protein with 12 putative PPR motifs that is located inside the chloroplast. The protein shows a significant structural, but not primary, sequence similarity to the maize protein CRP1, which is involved in the processing and translation of the chloroplast petD and petA RNAs. In addition, we found that HCF152 is an RNA binding protein that binds certain areas of the petB transcript. The protein possibly exists in the chloroplast as a homodimer and is not associated with other proteins to form a high molecular mass complex. PMID- 12782740 TI - Measurement quality and validity of the "need for recovery scale". AB - The "need for recovery scale" is suggested as an operationalisation for the measurement of (early symptoms of) fatigue at work. Definition of and background on the concept of need for recovery are briefly discussed. Details about scale construction are summarised. Correlations with other relevant measurement scales on fatigue at work are presented to validate the operationalisation claim, as are early results on predictive validity. A study is presented that further investigates the measurement quality and validity of the scale. The data used in this study were collected by Occupational Health Services for 68 775 workers during the period 1996-2000. Comparing the measurement quality of subgroups (Cronbach's alpha) differing in terms of age class, sex, and education level, the general applicability of the scale was shown. The validity of the scale was studied by analysing its association with psychosocial risk factors. Multiple regression analyses of need for recovery were performed on individual and department level data, using 10 psychosocial job characteristics as independent variables. The two most important factors in the explanation of variance at the individual level were also dominant at the department level: pace and amount of work, and emotional workload. The percentage of explained variance was higher at the department level than at the individual level, and increased with department size. Results suggest that the need for recovery scale is an adequate scale, both for applications at the individual and at the group (department/organisation) level. PMID- 12782741 TI - Assessment of fatigue among working people: a comparison of six questionnaires. AB - AIMS: To compare the psychometric qualities of six fatigue questionnaires in a sample of working persons. METHODS: Internal consistency and test-retest reliability, content validity, convergent validity, and the dimensionality of the fatigue instruments were explored. RESULTS: All scales had a satisfactory internal consistency. Furthermore, based on factor analyses and Mokken scale analyses, all scales were unidimensional and appeared to measure an identical construct. The Fatigue Assessment Scale (FAS) had the highest factor loading on the one factor solution obtained in a factor analysis of the total scores of all scales. CONCLUSIONS: All the questionnaires were unidimensional and had good reliability and validity. The FAS was the most promising fatigue measure. PMID- 12782742 TI - How to conduct research on burnout: advantages and disadvantages of a unidimensional approach in burnout research. AB - When conducting research on burnout, it may be difficult to decide whether one should report results separately for each burnout dimension or whether one should combine the dimensions. Although the multidimensionality of the burnout concept is widely acknowledged, for research purposes it is sometimes convenient to regard burnout as a unidimensional construct. This article deals with the question of whether and when it may be appropriate to treat burnout as a unidimensional variable, and presents a decision rule to distinguish between people high and low in burnout. To develop a guideline for obtaining a dichotomous measure of burnout, the scores on the Utrecht Burnout Scale (UBOS) of 44 well functioning individuals were compared with the scores of 29 individuals diagnosed as suffering from burnout. Based on these data, the authors recommend the "exhaustion + 1" criterion for research in non-clinical populations. Following this criterion, individuals can be considered as burnt out when they report, compared to a norm group, high emotional exhaustion, in combination with high depersonalisation or low personal accomplishment. The criterion may be used to estimate the percentage in a sample of individuals in a state of burnout. PMID- 12782743 TI - Quality of rehabilitation among workers with adjustment disorders according to practice guidelines; a retrospective cohort study. AB - AIMS: To assess the quality of occupational rehabilitation for patients with adjustment disorders and to determine whether high quality of care is related to a shorter period of sickness absence. METHODS: A retrospective cohort study was conducted by means of an audit of 100 files of patients with adjustment disorders who visited their occupational physicians. Quality of rehabilitation was assessed by means of 10 performance indicators, derived from the guidelines for the treatment of employees with mental health disorders. Performance was dichotomised into optimal and deviant care according to explicit criteria. The performance rates were related to time until work resumption during a one year follow up period. Kaplan-Meier survival analyses and Cox proportional hazards analysis were used to study this relation. RESULTS: Four of 10 performance rates were below 50%: continuity of care (34%), interventions aimed at providers of care in the curative sector (39%), assessment of impediments in the return to work process (41%), and assessment of symptoms (45%). The highest performance rate concerned assessment of work related causes (94%). Overall optimal care was found in 10% of the cases. Median time to complete recovery was 195 days (IQR 97 to 365), and 73% of all patients recovered completely after one year. Optimal continuity of care was significantly related to a shorter time to both partial and complete work resumption (hazard ratio (HR) 0.3; CI 0.2 to 0.6) independently of other performance indicators. Performance regarding interventions aimed at the organisation was also related to a shorter time until first return to work (HR 0.5; CI 0.3 to 0.9). CONCLUSIONS: This study shows that the rehabilitation process of employees with adjustment disorders leaves significant room for improvement, especially with regard to continuity of care. Quality of care was partly related to a better outcome. More rigorous study designs are needed to corroborate these findings. PMID- 12782744 TI - Fatigue, burnout, and chronic fatigue syndrome among employees on sick leave: do attributions make the difference? AB - BACKGROUND: Persistent fatigue among employees, burnout, and chronic fatigue syndrome (CFS) are three fatigue conditions that share some characteristics in theory. However, these conditions have not been compared in empirical research, despite conceptual similarities. METHODS: This cross sectional study aimed to investigate relations between persistent fatigue, burnout, and CFS by describing the clinical features of a sample of 151 fatigued employees on sick leave. Using validated instruments, subgroups based on research criteria for CFS and burnout within the sample of fatigued employees and a reference group of 97 diagnosed CFS patients were compared. Analyses of covariance were performed. RESULTS: A total of 66 (43.7%) fatigued employees met research criteria for CFS (except symptom criteria) and 76 (50.3%) met research criteria for burnout. "CFS-like employees" (fatigued employees who met CFS criteria) reported stronger somatic attributions than "non-CFS-like employees". Burnt out CFS-like employees were more depressed and distressed than CFS-like employees who were not burnt out. Burnout cases among the non-CFS-like employees had stronger psychological attributions than fatigued employees who were not burnt out. Compared to diagnosed CFS patients, CFS-like employees merely had a shorter duration of fatigue complaints. Burnt out CFS-like employees had stronger psychological attributions and were more distressed than CFS patients. CONCLUSIONS: Fatigued employees shared many important characteristics with CFS patients, regardless of burnout status, and many fatigued employees met CFS criteria and/or burnout criteria. Differences however concerned the causal attributions that were made. This raises questions about the role of causal attributions: are they modified by fatigue complaints or do they determine illness outcome? PMID- 12782745 TI - An epidemiological approach to study fatigue in the working population: the Maastricht Cohort Study. AB - In 1998, a large scale prospective cohort study of prolonged fatigue in the working population was started in the Netherlands. The ultimate goal of this Maastricht Cohort Study was to identify risk factors involved in the aetiology and natural course of prolonged fatigue in the working population and to develop preventive measures and treatments that can be used in occupational health settings. In this paper, a conceptual model for epidemiological research on prolonged fatigue is presented. This model is the basis for the Maastricht Cohort Study. Alongside the model and design, the characteristics of the study population, the prevalence and one year cumulative incidence of prolonged fatigue, as well as its relation with secondary health outcomes (psychological distress, need for recovery, and burnout) are presented. Furthermore, model, design, and the presented results are discussed. PMID- 12782746 TI - Acute and chronic job stressors among ambulance personnel: predictors of health symptoms. AB - OBJECTIVES: To predict symptomatology (post-traumatic distress, fatigue, and burnout) due to acute and chronic work related stressors among ambulance personnel. METHODS: Data were gathered from 123 ambulance workers in The Netherlands in a longitudinal design. At two measurements they completed standardised questionnaires to assess health symptoms, such as the Impact of Event Scale, the Maslach Burnout Inventory, and the Checklist Individual Strength. Acute stressors were assessed with specific questions, and chronic work related stressors were measured with the Questionnaire on the Experience and Assessment of Work. RESULTS: Most of the ambulance workers had been confronted with acute stressors in their work. They also reported more chronic work related stressors than a reference group. Of the participants, more than a tenth suffered from a clinical level of post-traumatic distress, a tenth reported a fatigue level that put them at high risk for sick leave and work disability and nearly a tenth of the personnel suffered from burnout. Best predictors of symptomatology at time 2 were lack of social support at work and poor communication, such as not being informed about important decisions within the organisation. CONCLUSIONS: Ambulance personnel are at risk to develop health symptoms due to work related stressors. Although, acute stressors are related to health symptoms, such as fatigue, burnout, and post-traumatic symptoms, it was not found to predict health symptoms in the long term. Main risk factors have to do with social aspects of the work environment, in particular lack of support from the supervisor as well as colleagues and poor communication. When implementing workplace interventions these social aspects need to be taken into account. PMID- 12782747 TI - Work schedules and fatigue: a prospective cohort study. AB - AIMS: (1) To describe the prevalence of fatigue among employees in different work schedules (day work, three-shift, five-shift, and irregular shift work); (2) to investigate whether different work schedules are related to increasing fatigue over time, while taking into account job title and job characteristics; and (3) to study fatigue among shift workers changing to day work. METHODS: Data from nine consecutive four-monthly self administered questionnaires from the Maastricht Cohort Study on Fatigue at work (n = 12 095) were used with 32 months of follow up. Day and shift workers were matched on job title. RESULTS: The prevalence of fatigue was 18.1% in day workers, 28.6% in three-shift, 23.7% in five-shift, and 19.1% in irregular shift workers. For three-shift and five-shift workers substantial higher fatigue levels were observed compared to day workers at baseline measurement. In the course of fatigue over the 32 months of follow up there were only small and insignificant differences between employees in different work schedules. However, among employees fatigued at baseline, fatigue levels decreased faster over time among five-shift workers compared to fatigued day workers. Shift workers changing to day work reported substantially higher fatigue levels prior to change, compared to those remaining in shift work. CONCLUSIONS: Substantial differences in fatigue existed between day and shift workers. However, as no considerable differences in the course of fatigue were found, these differences have probably developed within a limited time span after starting in a shift work job. Further, evidence was found that fatigue could be an important reason for quitting shift work and moving to day work. Finally, in the relation between work schedules and fatigue, perceived job characteristics might play an important role. PMID- 12782748 TI - Physiological differences between burnout patients and healthy controls: blood pressure, heart rate, and cortisol responses. AB - OBJECTIVES: To investigate differences between burnout patients and healthy controls regarding basal physiological values and physiological stress responses. Measures of the sympathetic-adrenergic-medullary (SAM) axis and the hypothalamic pituitary-adrenal (HPA) axis were examined. METHODS: SAM axis and HPA axis activity was compared between 22 burnout patients and 23 healthy controls. SAM axis activity was measured by means of heart rate (HR) and blood pressure (BP). HPA axis activity was investigated by means of salivary cortisol levels. Resting levels of HR, BP, and cortisol were determined as well as reactivity and recovery of these measures during a laboratory session involving mental arithmetic and speech tasks. In addition, morning levels of cortisol were determined. RESULTS: Burnout patients showed higher resting HR than healthy controls. BP resting values did not differ between burnout patients and healthy controls, nor did cardiovascular reactivity and recovery measurements during the laboratory session. Basal cortisol levels and cortisol reactivity and recovery measures were similar for burnout patients and healthy controls. However, burnout patients showed elevated cortisol levels during the first hour after awakening in comparison to healthy controls. CONCLUSIONS: The findings provided limited proof that SAM axis and HPA axis are disturbed among burnout patients. Elevated HR and elevated early morning cortisol levels may be indicative of sustained activation. PMID- 12782749 TI - Need for recovery from work related fatigue and its role in the development and prediction of subjective health complaints. AB - AIMS: To present the available empirical evidence for the assumed position of the concept of work related fatigue as: (1) short term effect of the working day; and (2) an intermediate variable between work demands and the development of subjective health complaints and sickness absence. METHODS: Results from six single occupation studies, conducted between 1996 and 2002, are presented. Work demands (working hours, decision latitude, break control/autonomy, and mental, emotional, and physical demands) were assessed through validated scales. Work related fatigue was represented and assessed by means of the need for recovery after working time scale in all studies. Subjective health complaints and duration of sickness absence were quantified with the same instruments in most studies as well. Both cross sectional studies (four) as well as prospective studies (two; up to two years follow up) were performed. Cross sectional data of 3820 workers, in total, were available. Prospective data were accessible for 1200 workers in industry and health care. Models were tested with stepwise multiple regression analyses. RESULTS: Strong associations between work demands and need for recovery were found in different occupations. The variance explained in need for recovery by work demands, age, and (baseline) need for recovery ranged between 14% and 48% in both types of studies. The amount of explained variance by work demands, age, and (baseline) need for recovery in subjective health complaints ranged between 24% and 58% in the different occupations. The prospective data showed the prognostic value of need for recovery in relation to subjective health complaints (in terms of psychosomatic complaints, emotional exhaustion, or sleep problems) and duration of future sickness absence. CONCLUSIONS: The hypothesised role for work related fatigue as a link in the causal string of events, that is assumed to exist between repeated adverse work demands and the development of work related stress reactions, (psychological) overload and, eventually, health problems, was confirmed. PMID- 12782750 TI - Fatigue as a predictor of sickness absence: results from the Maastricht cohort study on fatigue at work. AB - OBJECTIVES: To investigate whether there is a relationship between fatigue and sickness absence. Two additional hypotheses were based on the theoretical distinction between involuntary, health related absence and voluntary, attitudinal absence. In the literature, the former term is usually used to describe long term sickness absence, the latter relates to short term sickness absence. In line with this, the first additional hypothesis was that higher fatigue would correspond with a higher risk of long term, primarily health related absence. The second additional hypothesis was that higher fatigue would correspond with a higher risk of short term, primarily motivational absence. METHODS: A multidimensional fatigue measure, as well as potential sociodemographic and work related confounders were assessed in the baseline questionnaire of the Maastricht cohort study on fatigue at work. Sickness absence was objectively assessed on the basis of organisational absence records and measured over the six months immediately following the baseline questionnaire. In the first, general hypothesis the effect of fatigue on time-to-onset of first sickness absence spell during follow up was investigated. For this purpose, a survival analysis was performed. The effect of fatigue on long term sickness absence was tested by a logistic regression analysis. The effect of fatigue on short term sickness absence was investigated by performing a survival analysis with time-to-onset of first short absence spell as an outcome. RESULTS: It was found that higher fatigue decreased the time-to-onset of the first sickness absence spell. Additional analyses showed that fatigue was related to long term as well as to short term sickness absence. The effect of fatigue on the first mentioned outcome was stronger than the effect on the latter outcome. Potential confounders only weakened the effect of fatigue on long term absence. CONCLUSIONS: Fatigue was associated with short term but particularly with long term sickness absence. The relation between fatigue and future sickness absence holds when controlling for work related and sociodemographic confounders. Fatigue as measured with the Checklist Individual Strength can be used as a screening instrument to assess the likelihood of sickness absence in the short term. PMID- 12782751 TI - The Depression Anxiety Stress Scales (DASS): detecting anxiety disorder and depression in employees absent from work because of mental health problems. AB - AIMS: To (1) evaluate the psychometric properties and (2) examine the ability to detect cases with anxiety disorder and depression in a population of employees absent from work because of mental health problems. METHODS: Internal consistency, construct validity, and criterion validity of the Depression Anxiety Stress Scales (DASS) were assessed. Furthermore, the ability to identify anxiety disorders or depression was evaluated by calculating posterior probabilities of these disorders following positive and negative test results for different cut off scores of the DASS-Depression and DASS-Anxiety subscales. RESULTS: Internal consistency of the DASS subscales was high, with Cronbach's alphas of 0.94, 0.88, and 0.93 for depression, anxiety, and stress respectively. Factor analysis revealed a three factor solution, which corresponded well with the three subscales of the DASS. Construct validity was further supported by moderately high correlations of the DASS with indices of convergent validity (0.65 and 0.75), and lower correlations of the DASS with indices of divergent validity (range -0.22 to 0.07). Support for criterion validity was provided by a statistically significant difference in DASS scores between two diagnostic groups. A cut off score of 5 for anxiety and 12 for depression is recommended. The DASS showed probabilities of anxiety and depression after a negative test result of 0.05 and 0.06 respectively. Probabilities of 0.29 for anxiety disorder and 0.33 for depression after a positive test result reflect relatively low specificity of the DASS. CONCLUSION: The psychometric properties of the DASS are suitable for use in an occupational health care setting. The DASS can be helpful in ruling out anxiety disorder and depression in employees with mental health problems. PMID- 12782752 TI - Need for recovery after work and the subsequent risk of cardiovascular disease in a working population. AB - BACKGROUND: A high need for recovery after work can be regarded as a short term adverse effect of working day stressors and the person's inability to cope and recover. Consequently, it might be an intermediate factor between job stressors and cardiovascular disease (CVD). AIM: To investigate, in a longitudinal study, the relation between need for recovery and subsequent CVD. METHODS: Data from the Maastricht Cohort Study of 12 140 workers were used, with 42 incident self reported CVD cases during 32 months of follow up. Cox proportional hazards analysis was used to calculate age, gender, smoking status, and educational level adjusted relative risks. RESULTS: The adjusted relative CVD risk for the second compared to the first tertile of the need for recovery score was 1.22 (95% CI: 0.49 to 3.04), and for the third compared to the first tertile was 3.16 (95% CI: 1.34 to 7.48). When need for recovery was entered as continuous score, an adjusted relative risk per SD increase of 1.54 (95% CI: 1.15 to 2.03) was found. Additional adjustment for several work related factors as job demands, did not notably change the observed relation between need for recovery and CVD. Moreover, the increased risk for subjects reporting high job demands (1.38 per SD increase; 95% CI: 1.02 to 3.92) decreased substantially after adjustment for need for recovery. CONCLUSION: The results show that need for recovery is a strong predictor of subsequent cardiovascular disease and might be an intermediate factor between job stressors and cardiovascular disease. PMID- 12782753 TI - Fatigue as a risk factor for being injured in an occupational accident: results from the Maastricht Cohort Study. AB - AIMS: To determine whether fatigue and need for recovery are risk factors for being injured in an occupational accident. METHODS: These associations were investigated within the Maastricht Cohort Study of "Fatigue at Work", a prospective cohort study of employees from a wide range of companies and organisations. For 7051 employees information was available on fatigue as measured with the Checklist Individual Strength (CIS), need for recovery as measured with the VBBA, and possible confounding factors such as age, smoking, alcohol consumption, educational level, shift work, and work environment. Information on the risk factors was collected in May 1999 and January 2000, before the occurrence of the occupational accidents. The incidence of being injured in an occupational accident was inventoried over the year 2000. A total of 108 employees reported having been injured in an occupational accident in 2000. RESULTS: For the highest CIS fatigue score tertile a for age, gender, educational level, smoking, shift work, and work environment, adjusted relative risk for being injured in an occupational accident of 1.29 (95% CI: 1.03 to 2.78) was found compared to the lowest tertile, and for the highest tertile of need for recovery a relative risk of 2.28 (95% CI: 1.41 to 3.66) was found. CONCLUSIONS: Fatigue and need for recovery were found to be independent risk factors for being injured in an occupational accident. This means that in the push back of occupational accidents, fatigue, and even more importantly need for recovery, need special attention. PMID- 12782754 TI - Fatigue in employees with diabetes: its relation with work characteristics and diabetes related burden. AB - AIMS: To examine the relations between work characteristics as defined by the Job Demand-Control-Support model (JDCS) (that is, job demands, decision latitude, and social support), diabetes related burden (symptoms, seriousness of disease, self care activities, and disease duration), and fatigue in employees with diabetes mellitus. METHODS: Employees (n = 292) aged 30-60 years, with insulin treated diabetes, filled in self administered questionnaires that assess the above mentioned components of the JDCS model and diabetes related burdens. RESULTS: Both work and diabetes related factors are related to fatigue in employees with diabetes. Regression analyses revealed that work characteristics explain 19.1% of the variance in fatigue; lack of support, and the interaction of job demands and job control contribute significantly. Diabetes related factors explain another 29.0% of the variance, with the focus on diabetes related symptoms and the burden of adjusting insulin dosage to circumstances. Fatigue is more severe in case of lack of social support at work, high job demands in combination with a lack of decision latitude, more burden of adjusting insulin dosage to circumstances, and more diabetic symptoms. Furthermore, regression analysis revealed that diabetic symptoms and the burden of adjusting the insulin dosage to circumstances are especially relevant in combination with high job demands. CONCLUSIONS: Both diabetes and work should be taken into consideration-by (occupational) physicians as well as supervisors-in the communication with people with diabetes. PMID- 12782755 TI - Associations between fatigue attributions and fatigue, health, and psychosocial work characteristics: a study among employees visiting a physician with fatigue. AB - AIMS: To study associations between characteristics of employees active at work and making a fatigue related visit to the general practitioner (GP) or occupational physician (OP) in terms of fatigue, physical health problems, mental health problems, psychosocial work characteristics, and attributions of their fatigue complaints. METHODS: Self report questionnaires from the Maastricht Cohort Study Fatigue at Work were used to measure fatigue (Checklist Individual Strength, Maslach Burnout Inventory-General Survey), physical health problems (chronic illness), mental health problems (Hospital Anxiety and Depression Scale), psychosocial work characteristics (Job Content Questionnaire), and fatigue attributions (somatic, psychological, none) in employees who made a fatigue related visit to the GP or OP over a six month period. RESULTS: In employees visiting only the GP, fatigue was an important reason to visit in one of seven (13.9%) employees. These fatigue related visits were in particular associated with high fatigue levels and mental health problems. A psychological fatigue attribution was reported by 41.8%, a somatic fatigue attribution by 44.0%. On a multivariate level, mental health problems showed the strongest association with psychological fatigue attributions, over and beyond fatigue itself. No associations were found between fatigue attributions and psychosocial work characteristics. Attributional patterns appeared to be different between visitors of the GP and the OP. CONCLUSIONS: Fatigue is a common reason among employees to consult a GP. Asking employees for their own fatigue attributions in terms of somatic or psychological causes may be useful for the GP-and possibly also the OP-to gather information about underlying health problems in employees active at work and making a fatigue related visit. PMID- 12782757 TI - Sonic hedgehog. PMID- 12782758 TI - Genetic vulnerability following traumatic brain injury: the role of apolipoprotein E. AB - Apolipoprotein E (APOE) is thought to be responsible for the transportation of lipids within the brain, maintaining structural integrity of the microtubule within the neurone, and assisting with neural transmission. Possession of the APOE epsilon4 allele has also been shown to influence neuropathological findings in patients who die from traumatic brain injury, including the accumulation of amyloid beta protein. Previous clinical studies reporting varying outcome severities of traumatic brain injury, including cognitive and functional recovery, all support the notion that APOE epsilon4 allele possession is associated with an unfavourable outcome. Evidence from experimental and clinical brain injury studies confirms that APOE plays an important role in the response of the brain to injury. PMID- 12782759 TI - Molecular changes in the Ki-ras and APC genes in primary colorectal carcinoma and synchronous metastases compared with the findings in accompanying adenomas. AB - AIMS: To compare the molecular genetic changes in the Ki-ras and adenomatous polyposis coli (APC) genes between colorectal carcinomas and synchronous metastases, and then to compare and contrast those changes with previously reported changes in the two genes between these carcinomas and accompanying adenomas. This expanded comparison would provide greater understanding of the progression of molecular changes in neoplastic tissue during the development of malignancy from a benign adenoma to carcinoma and then to metastatic spread of the malignancy. METHODS: DNA was extracted from paraffin wax embedded tissue. This was followed by polymerase chain reaction and gel electrophoresis for mutations in the Ki-ras gene using single stranded conformational polymorphism analysis. Amplification of a CA repeat marker was used to assess loss of heterozygosity (LOH) at the APC gene. RESULTS: The findings for the Ki-ras gene in 42 paired carcinomas and synchronous metastases were identical, regardless of whether or not the carcinoma and its companion adenoma had identical Ki-ras findings. The results of APC LOH for 39 paired carcinomas and synchronous metastases were also identical, whether or not the carcinoma and its companion adenoma had identical APC LOH findings. Results were uninformative for three pairs. CONCLUSIONS: With respect to these two genes, a carcinoma may be discordant from its companion adenoma, but the metastasis remains consistent with the colonic carcinoma. PMID- 12782760 TI - Regions of allelic imbalance in the distal portion of chromosome 12q in gastric cancer. AB - AIMS: To define regions of loss on the distal portion of chromosome 12q in gastric adenocarcinoma. METHODS: Microsatellite analysis on chromosome 12 was performed on 19 human gastric cancer cell lines using 77 markers, 71 of which were within or distal to 12q21; some portions of this region showed extended regions of homozygosity (ERHs) in 10 of 19 gastric cancer cell lines. In addition, microdissected tumour cells from 76 primary gastric adenocarcinomas were examined using 13 markers of interest implicated by the cell line data; 70% of these showed allelic imbalance (AI) at one or more markers in or distal to 12q21. RESULTS: Mapping ERHs in the cell lines and sites of AI in the tumours identified three regions that contain putative tumour suppressor genes: region A is located within 2.8 Mb between markers D12S1667 and D12S88; region B, within 1.9 Mb between markers D12S1607 and D12S78; and region C, in 0.74 Mb between markers D12S342 and D12S324. Fluorescence in situ hybridisation (FISH) analysis in two cell lines confirmed that two of the ERHs reflected deletions, not amplifications, of D12S81 in region A and D12S340 in region C. FISH analysis of marker D12S1075 within an ERH containing region B in one cell line showed neither amplification nor deletion. AI on 12q was not associated with prognosis, but was associated with ethnicity of the patient. CONCLUSIONS: These results identify regions on chromosome 12 that appear to contain tumour suppressor genes important in the development of gastric cancer. PMID- 12782762 TI - Epstein-Barr virus encoded latent membrane protein 1 (LMP1) and TNF receptor associated factors (TRAF): colocalisation of LMP1 and TRAF1 in primary EBV infection and in EBV associated Hodgkin lymphoma. AB - AIMS: Epstein-Barr virus (EBV) immortalises B cells in vitro and is associated with several malignancies. Most phenotypic effects of EBV are mediated by latent membrane protein 1 (LMP1), which interacts with tumour necrosis factor receptor associated factors (TRAFs) to activate NF-kappaB. This study examines TRAF1 and LMP1 expression in EBV associated lymphoproliferations. METHODS: TRAF1 expression was investigated in 26 Hodgkin lymphomas (HL; 18 EBV+, eight EBV-), seven EBV+ Burkitt lymphomas (BL), two infectious mononucleosis (IM) tonsils, and lymphoreticular tissue from eight chronic virus carriers. Seven anaplastic large cell lymphomas and 10 follicular B cell lymphomas were also studied. Colocalisation of TRAF1 and LMP1 was studied by immunofluorescent double labelling and confocal laser microscopy. RESULTS: TRAF1 colocalises with LMP1 in EBV infected cells in IM. EBV positive lymphocytes from chronic virus carriers were negative for TRAF1 and LMP1. In HL biopsies, TRAF1 was strongly expressed independently of EBV status, whereas all BL cases were TRAF1-. In EBV+ HL cases, TRAF1 colocalised with LMP1. Eight of 10 follicular lymphomas expressed TRAF1 in centroblast-like cells. Four of seven anaplastic large cell lymphomas weakly expressed TRAF1. CONCLUSIONS: These results suggest that in non-neoplastic lymphocytes, TRAF1 expression is dependent on the presence of LMP1, and that in IM B cells in vivo, LMP1 associated signalling pathways are active. In HL, TRAF1 is expressed independently of EBV status, probably because of constitutive NF kappaB activation. The function of TRAF1 in HL remains to be determined. PMID- 12782763 TI - Measurement of thyroglobulin mRNA in peripheral blood as an adjunctive test for monitoring thyroid cancer. AB - AIMS: Monitoring treated patients with thyroid cancer for recurrent or metastatic disease is currently based upon the serial measurement of circulating plasma thyroglobulin (Tg) concentrations. However, the clinical usefulness of Tg immunoassays is limited by poor sensitivity and interference from anti-Tg antibodies. This study investigated whether the detection of Tg mRNA in peripheral blood, using reverse transcriptase polymerase chain reaction (RT-PCR), is of value in the biochemical surveillance of patients with thyroid cancer. METHODS: RNA was extracted from peripheral blood of five normal controls, six patients with abnormal thyroid function tests, and 28 patients who had undergone thyroidectomy for well differentiated thyroid cancer. From each, an 87 bp product from base pair 262 to 348 in the cDNA sequence of the thyroglobulin gene was amplified by RT-PCR. RESULTS: Tg mRNA was detected in normal individuals and patients with thyroid cancer. In the group of patients studied, identification of metastatic thyroid tissue by radioiodine scanning correlated better with Tg mRNA assay results than with serum Tg concentrations (accuracy 84% v 75%). No interference from circulating Tg antibodies was apparent. In patients studied prospectively over a 12 month period, there was a significant correlation between detectable Tg mRNA in peripheral blood and the presence or absence of metastatic disease, as demonstrated by radioiodine scanning. CONCLUSIONS: These results suggest that detection of Tg mRNA in blood is a more sensitive marker for metastatic thyroid disease than Tg immunoassay, and appears to be unaffected by the presence of circulating anti-Tg antibodies. PMID- 12782764 TI - Increased zinc finger protein zFOC1 transcripts in gastric cancer compared with normal gastric tissue. AB - BACKGROUND: Screening of cDNA arrays of the IMAGE library identified human zFOC1 as a differentially expressed cDNA that was upregulated in KATO III gastric cancer cells following stimulation with the gastric pathogen Helicobacter pylori. AIMS: To determine the expression of zFOC1 in gastric mucosa with and without H pylori infection and in patients with gastric cancer. RESULTS: zFOC1 is localised on chromosome 12q24.3 and encodes a zinc finger protein. Expression studies in human H pylori infected and uninfected gastric biopsies, gastric tumours, and gastric cancer cell lines revealed that zFOCI gene transcripts are significantly higher in gastric cancer than in non-cancerous gastric tissues. CONCLUSIONS: The zFOC1 gene appears to be a tumour marker associated with gastric cancer. PMID- 12782766 TI - Three novel PAX6 mutations in patients with aniridia. AB - AIMS: To describe mutations in the PAX6 gene in five patients with aniridia from three unrelated families. METHODS: The PAX6 gene was analysed using single stranded conformational polymorphism analysis and direct sequencing. RESULTS: In one family, three individuals from two generations had aniridia, whereas in each of the other families only one member was affected. The first patient had the heterozygous Q221X (1023C --> T) nonsense mutation in exon 8. The same mutation was found in his mother and sister. Another patient had a heterozygous Q297X (1252C --> T) mutation in exon 10. The third patient carried a heterozygous IVS5+2T --> C mutation leading to aberrant splicing of mRNA. CONCLUSIONS: These findings provide further examples of haploinsufficiency of PAX6 in aniridia. PMID- 12782767 TI - PCR analysis in archival postmortem tissues. AB - BACKGROUND: Formalin fixed and paraffin wax embedded tissues of necropsy origin are an important source for molecular analysis especially in rare diseases, neuropathology, or molecular epidemiology studies. Because of DNA degradation, only short sequences can be amplified from this type of tissue, very often less than 100 bases. This poses problems because studies on polymorphism and mutations occurring in large genes often require the analysis of long sequences. METHODS: The development of a simple treatment to obtain longer fragments of DNA for the analysis of archival postmortem paraffin wax embedded tissues. RESULTS: It was possible to amplify longer sequences ranging up to 300 bases from postmortem tissues, with no modification to the usual DNA extraction procedures. To obtain longer stretches of DNA, a pre-PCR restoration treatment was required, by filling single strand breaks, followed by a vigorous denaturation step. CONCLUSIONS: The development of this simple treatment allowed the analysis of longer fragments of DNA obtained from archival postmortem paraffin wax embedded tissues. PMID- 12782768 TI - No evidence of tumour cells in blood of patients with glioma. AB - BACKGROUND: Although clinically apparent systemic metastases of gliomas are very rare, reports of gliomas developing in recipient's transplanted organs have suggested that haematogenous spread might be more common. METHODS: This report describes a newly developed, sensitive real time quantitative reverse transcription polymerase chain reaction assay for the detection of mRNA encoding glial fibrillary acidic protein (GFAP). Blood from 10 patients with astrocytoma and 10 patients with glioblastoma was analysed. RESULTS: No GFAP mRNA was detected. CONCLUSIONS: These results suggest that even subclinical metastases are very rare and are probably restricted to distinct subsets of glioma. PMID- 12782769 TI - Uses and abuses of genetic engineering. PMID- 12782770 TI - Occupation and gastric cancer. AB - Gastric cancer is a cause of significant morbidity and mortality. There are several risk factors, with occupation emerging as one of these. There is considerable evidence that occupations in coal and tin mining, metal processing, particularly steel and iron, and rubber manufacturing industries lead to an increased risk of gastric cancer. Other "dusty" occupations-for example, wood processing, or work in high temperature environments have also been implicated but the evidence is not strong. The mechanism of pathogenesis of gastric cancer is unclear and the identification of causative agents can be difficult. Dust is thought to be a contributor to the pathological process, but well known carcinogens such as N-nitroso compounds have been detected in some environments. Further research on responsible agents is necessary and screening for detection of precursor gastric cancer lesions at the workplace merits consideration. PMID- 12782772 TI - Mechanisms and management of diuretic resistance in congestive heart failure. AB - Diuretic drugs are used almost universally in patients with congestive heart failure, most frequently the potent loop diuretics. Despite their unproven effect on survival, their indisputable efficacy in relieving congestive symptoms makes them first line therapy for most patients. In the treatment of more advanced stages of heart failure diuretics may fail to control salt and water retention despite the use of appropriate doses. Diuretic resistance may be caused by decreased renal function and reduced and delayed peak concentrations of loop diuretics in the tubular fluid, but it can also be observed in the absence of these pharmacokinetic abnormalities. When the effect of a short acting diuretic has worn off, postdiuretic salt retention will occur during the rest of the day. Chronic treatment with a loop diuretic results in compensatory hypertrophy of epithelial cells downstream from the thick ascending limb and consequently its diuretic effect will be blunted. Strategies to overcome diuretic resistance include restriction of sodium intake, changes in dose, changes in timing, and combination diuretic therapy. PMID- 12782773 TI - Paediatric tuberculosis. AB - Children are important in the epidemiology of tuberculosis as a marker of recent disease transmission and a reservoir for the future. Once infected they have a higher risk of progressing to tuberculous disease. Chest radiography and tuberculin testing with or without tissue for culture are still the standard tools for confirming the diagnosis once this is considered. Well researched treatment protocols are available but multidrug resistant tuberculosis and coexistent HIV are a challenge. Ensuring compliance with treatment is a major concern. Controversy still surrounds the place of BCG. Advances in the molecular genetics of tuberculosis hold out the possibility of better vaccines. PMID- 12782771 TI - Management of asthma in adults: current therapy and future directions. AB - Asthma is increasing in prevalence worldwide and results in significant use of healthcare resources. Although most patients with asthma can be adequately treated with inhaled corticosteroids, an important number of patients require additional therapy and an increasing number of options are available. A further minority of patients develop severe persistent asthma which remains difficult to manage despite current pharmacological therapies. This review discusses the various treatment options currently available for each stage of asthma severity, highlights some of the limitations of current management, and outlines directions which may improve the management of asthma in the future. PMID- 12782774 TI - Iatrogenic harlequin syndrome. PMID- 12782775 TI - Comparative study of bone mineral density, calcium, and vitamin D status in the Gujarati and white populations of Leicester. AB - OBJECTIVES: To evaluate differences in bone mineral density (BMD), calcium, and vitamin D status between the Gujarati (South Asian) and white populations resident in Leicester and to determine whether this was linked to lifestyle factors. DESIGN: An observational cross sectional study of randomly selected Gujarati and white volunteers aged from 20-40 years. SETTING: City of Leicester. PARTICIPANTS: Subjects were randomly selected by age (20-40 years) and ethnicity. A total of 262 individuals volunteered to participate, of which 201 (51 white females, 71 Gujarati females, 37 white males, 42 Gujarati males) were eligible for the study. MAIN OUTCOME MEASURES: Results of questionnaire, BMD at the hip and lumbar spine, and measurement of serum calcium, albumin, alkaline phosphatase, and 25-hydroxyvitamin D. RESULTS: Male and female white subjects were significantly taller and heavier than their Gujarati counterparts. There was a statistically significant difference in BMD both at the spine (p<0.001) and hip (p<0.001) between the white and Gujarati females with the Gujaratis having a lower BMD. There was a trend for Gujarati males to have a lower BMD at the hip and spine than their white counterparts but these figures did not reach statistical significance. The intensity of cigarette smoking and the amount of alcohol consumption were both higher in the white male and female subjects. Sunlight exposure (>4 hours per day) was significantly higher in white subjects compared with Gujaratis. There were no significant differences in the mean level of serum calcium or alkaline phosphatase between the Gujaratis and whites. A significantly higher proportion (p<0.001) of the Gujarati men and women had a vitamin D level that was not measurable (that is, below the lower limit of the laboratory range of normal). Of those who had a measurable level (that is, in the normal range) mean levels of vitamin D were lower (p<0.05) in the Gujarati men and women. CONCLUSION: The present study is the first of its kind to note a low BMD in Gujarati subjects of South Asian origin compared with their white counterparts, living in Leicester. This study also confirms the presence of low serum vitamin D levels in Gujaratis. There is a need for more research in South Asians with regard to the collection of normal BMD values. This could provide a more meaningful reference range for identifying South Asians at risk of osteoporotic fractures and may have public health implications of relevance to this ethnic group. PMID- 12782776 TI - Feasibility and safety of day care laparoscopic cholecystectomy in a developing country. AB - BACKGROUND: Although day care laparoscopic cholecystectomy (DCLC) has been shown to be safe in centres with adequate infrastructure for day care surgery, its feasibility and safety in developing countries has never been studied. Because of differences in the quality of health care delivery, western guidelines for day care surgery cannot be universally applied to developing countries. PATIENTS AND METHODS: Patients less than 65 years who were graded I and II on the American Society of Anesthesiologists physical status score, irrespective of their educational status, living within 20 km, and willing to make their own arrangements for a return to hospital in case of problems were selected for DCLC. Follow up was done by patients calling the hospital the morning after surgery. RESULTS: 50% of the eligibility criteria were new; 313/383 patients were suitable for DCLC. The commonest cause for rejection was that the patient lived out of the defined area (50%). Altogether 92% were discharged within eight hours of surgery. The reasons for failure to discharge were the presence of abdominal drains in four (2%), nausea and vomiting in nine (3%), and conversion to open surgery in five (2%). Ten patients (3%) were readmitted; of these only two (<1%) had complications needing re-exploration. Analysis of results showed that the inclusion and discharge criteria were valid and that the readmission and complication rates as well as the ease and accuracy of follow up were comparable to published data. DCLC reduced waiting times and increased patient turnover and may have a positive impact on resident training. CONCLUSIONS: DCLC is safe, feasible, and has potential benefits for health care delivery in developing countries. Each surgical service needs to develop their own guidelines based on local patient demography. PMID- 12782777 TI - Assessment of basic surgical trainees: can we do more? AB - INTRODUCTION: Changes to surgical training and reduction of junior doctors' working hours has resulted in trainees spending less time in surgical specialties before becoming eligible to apply for specialist registrar posts. A high quality basic surgical training programme is needed to improve the competence of trainees during their shortened period, an essential part of which is to conduct a formal assessment at the end of each training post and regular appraisals during their period of training. AIMS: To analyse the existing practice with regard to conducting assessment and appraisals for basic surgical trainees in the South East Wales region. METHODS: A questionnaire was sent to all the basic surgical trainees in the South East Wales region. RESULTS: A total of 52 questionnaires were sent out and 44 (84%) were returned. Four candidates were in the first post of their rotation, and were therefore excluded from the study. Nine of the 40 trainees (23%) did not have an assessment in one or more of their posts. There are 17 senior house officer posts available in general surgery in the rotation. Of the 31 "six month" episodes spent in these 17 posts, 27 (87%) underwent an assessment, 12 of the 17 (70%) episodes in trauma and orthopaedics (12 available posts) were assessed, and 31 of the 35 (88%) episodes in accident and emergency (eight available posts) underwent an assessment. Twenty eight of the 40 trainees (70%) did not undergo regular appraisals during the majority of their posts. Only 30%-50% of episodes spent in general surgery, orthopaedics, and accident and emergency underwent appraisals. The majority of the episodes spent in the other surgical specialties (12 posts available in total) had formal assessment and regular appraisals except for urology in which two of seven episodes underwent assessment and one underwent an appraisal. Twelve candidates (30%) expressed the view that operative experience and technical skills must form part of their assessment. Eight trainees (20%) felt that they needed more help from tutors with regard to career advice, preparation of curriculum vitae, and interview techniques for specialist registrar posts. Eight trainees mentioned that they had discussed unsatisfactory posts with their tutors but were not aware of any changes made to these posts. CONCLUSIONS: This study has shown that the formal assessment of basic surgical trainees at the end of each training post has to improve further. The existing practice of conducting regular appraisals for the trainees is grossly inadequate. There appears to be a need for formulating guidelines for conducting such appraisals, which would help to improve practice in the future. PMID- 12782778 TI - How valuable is ascitic cytology in the detection and management of malignancy? AB - INTRODUCTION: Ascitic cytology is often requested in the early stages of ascitic assessment. A review of this practice in a major English teaching hospital is presented. METHOD: Patients were retrospectively identified using the histopathology and patient administration system between January 1999 and May 2001. RESULTS: Of 276 samples sent for assessment 35 cases were found to be negative when on further review an intra-abdominal malignancy was present. The malignancy was diagnosed using a radiological modality. The sensitivity of ascitic cytology was found to be 60% with 100% specificity. A delay of up to five days could be incurred awaiting the cytology results before further radiological examinations were undertaken. CONCLUSION: Too much hope is placed on ascitic cytology to provide the diagnosis at the expense of other investigations. It is recommended that the initial assessment should concentrate on history, examination, and basic tests on ascitic fluid to assess the serum-ascites albumin gradient. Ovarian malignancy is the only tumour type yielding a significant rate of detection from cytology with some prognostic impact. Results should not be awaited before abdominal ultrasound is undertaken. This more directed practice would help reduce unnecessary workload for the pathologist and has resource implications. PMID- 12782779 TI - Recurrent aseptic meningitis due to different non-steroidal anti-inflammatory drugs including rofecoxib. AB - Aseptic meningitis can be caused by viruses, drugs, and connective tissue disorders. The most common drugs causing it include antibiotics like trimethoprim sulfamethoxazole, non-steroidal anti-inflammatory drugs (NSAIDs), intravenous immunoglobulins, intrathecal agents, vaccines, and monoclonal antibodies. A patient who had aseptic meningitis from three different NSAIDs including rofecoxib is presented. PMID- 12782780 TI - Ventricular fibrillation during electrical cardioversion of pre-excited atrial fibrillation. AB - The Wolff-Parkinson-White syndrome can rarely present with pre-excited atrial fibrillation. In this condition the short refractory period of the accessory pathway can lead to rapid atrioventricular conduction. There is then a danger that at high heart rates the irregular broad complex tachycardia that results can deteriorate into ventricular fibrillation. The initial management of patients presenting in pre-excited atrial fibrillation requires cardioversion to sinus rhythm. This can be performed by DC cardioversion or pharmacological means. This paper describes the case of a patient presenting in pre-excited atrial fibrillation where electrical DC cardioversion lead to transient iatrogenic ventricular fibrillation. PMID- 12782781 TI - A complication of hip surgery. PMID- 12782782 TI - Mass in the ear canal. PMID- 12782785 TI - NMR structure of the active conformation of the Varkud satellite ribozyme cleavage site. AB - Substrate cleavage by the Neurospora Varkud satellite (VS) ribozyme involves a structural change in the stem-loop I substrate from an inactive to an active conformation. We have determined the NMR solution structure of a mutant stem-loop I that mimics the active conformation of the cleavage site internal loop. This structure shares many similarities, but also significant differences, with the previously determined structures of the inactive internal loop. The active internal loop displays different base-pairing interactions and forms a novel RNA fold composed exclusively of sheared G-A base pairs. From chemical-shift mapping we identified two Mg2+ binding sites in the active internal loop. One of the Mg2+ binding sites forms in the active but not the inactive conformation of the internal loop and is likely important for catalysis. Using the structure comparison program mc-search, we identified the active internal loop fold in other RNA structures. In Thermus thermophilus 16S rRNA, this RNA fold is directly involved in a long-range tertiary interaction. An analogous tertiary interaction may form between the active internal loop of the substrate and the catalytic domain of the VS ribozyme. The combination of NMR and bioinformatic approaches presented here has identified a novel RNA fold and provides insights into the structural basis of catalytic function in the Neurospora VS ribozyme. PMID- 12782786 TI - Crystalline monoclonal antibodies for subcutaneous delivery. AB - Therapeutic applications for mAbs have increased dramatically in recent years, but the large quantities required for clinical efficacy have limited the options that might be used for administration and thus have placed certain limitations on the use of these agents. We present an approach that allows for s.c. delivery of a small volume of a highly concentrated form of mAbs. Batch crystallization of three Ab-based therapeutics, rituximab, trastuzumab, and infliximab, provided products in high yield, with no detectable alteration to these proteins and with full retention of their biological activity in vitro. Administration s.c. of a crystalline preparation resulted in a remarkably long pharmacokinetic serum profile and a dose-dependent inhibition of tumor growth in nude mice bearing BT 474 xenografts (human breast cancer cells) in vivo. Overall, this approach of generating high-concentration, low-viscosity crystalline preparations of therapeutic Abs should lead to improved ease of administration and patient compliance, thus providing new opportunities for the biotechnology industry. PMID- 12782787 TI - Expression profiles of acute lymphoblastic and myeloblastic leukemias with ALL-1 rearrangements. AB - The ALL-1 gene is directly involved in 5-10% of acute lymphoblastic leukemias (ALLs) and acute myeloid leukemias (AMLs) by fusion to other genes or through internal rearrangements. DNA microarrays were used to determine expression profiles of ALLs and AMLs with ALL-1 rearrangements. These profiles distinguish those tumors from other ALLs and AMLs. The expression patterns of ALL-1 associated tumors, in particular ALLs, involve oncogenes, tumor suppressors, antiapoptotic genes, drug-resistance genes, etc., and correlate with the aggressive nature of the tumors. The genes whose expression differentiates between ALLs with and without ALL-1 rearrangement were further divided into several groups, enabling separation of ALL-1-associated ALLs into two subclasses. One of the groups included 43 genes that exhibited expression profiles closely linked to ALLs with ALL-1 rearrangements. Further, there were evident differences between the expression profiles of AMLs in which ALL-1 had undergone fusion to other genes and AMLs with partial duplication of ALL-1. The extensive analysis described here pinpointed genes that might have a direct role in pathogenesis. PMID- 12782788 TI - Allele-specific silencing of dominant disease genes. AB - Small interfering RNA (siRNA) holds therapeutic promise for silencing dominantly acting disease genes, particularly if mutant alleles can be targeted selectively. In mammalian cell models we demonstrate that allele-specific silencing of disease genes with siRNA can be achieved by targeting either a linked single-nucleotide polymorphism (SNP) or the disease mutation directly. For a polyglutamine neurodegenerative disorder in which we first determined that selective targeting of the disease-causing CAG repeat is not possible, we took advantage of an associated SNP to generate siRNA that exclusively silenced the mutant Machado Joseph disease/spinocerebellar ataxia type 3 allele while sparing expression of the WT allele. Allele-specific suppression was accomplished with all three approaches currently used to deliver siRNA: in vitro-synthesized duplexes as well as plasmid and viral expression of short hairpin RNA. We further optimized siRNA to specifically target a missense Tau mutation, V337M, that causes frontotemporal dementia. These studies establish that siRNA can be engineered to silence disease genes differing by a single nucleotide and highlight a key role for SNPs in extending the utility of siRNA in dominantly inherited disorders. PMID- 12782789 TI - Retinoid activation of retinoic acid receptor but not retinoid X receptor is sufficient to rescue lethal defect in retinoic acid synthesis. AB - Two isomers of retinoic acid (RA) may be necessary as ligands for retinoid signaling: all-trans-RA for RA receptors (RARs) and 9-cis-RA for retinoid X receptors (RXRs). This was explored by using retinaldehyde dehydrogenase (Raldh)2 /- mouse embryos lacking mesodermal RA synthesis that display early growth arrest unless rescued by all-trans-RA administration. Because isomerization of all-trans RA to 9-cis-RA can occur, it is unclear whether both ligands are needed for rescue. We show here that an RAR-specific ligand can rescue Raldh2-/- embryos as efficiently as all-trans-RA, whereas an RXR-specific ligand has no effect. Further, whereas all-trans-RA was detected in embryos, 9-cis-RA was undetectable unless a supraphysiological dose of all-trans-RA was administered, revealing that 9-cis-RA is of pharmacological but not physiological significance. Because 9-cis RA is undetectable and unnecessary for Raldh2-/- rescue, and others have shown that 4-oxo-RA is unnecessary for mouse development, all-trans-RA emerges as the only ligand clearly necessary for retinoid receptor signaling. PMID- 12782790 TI - Microangiectasias: structural regulators of lymphocyte transmigration. AB - The migration of lymphocytes into inflammatory tissue requires the migrating cell to overcome mechanical forces produced by blood flow. A generally accepted hypothesis is that these forces are overcome by a multistep sequence of adhesive interactions between lymphocytes and endothelial cells. This hypothesis has been recently challenged by results demonstrating wall shear stress on the order of 20 dyn/cm(2) in vivo and infrequent lymphocyte-endothelial adhesion at wall shear stress >1-2 dyn/cm(2) in vitro. Here, we show that lymphocyte slowing and transmigration in the skin is associated with microangiectasias, i.e., focal structural dilatations of microvessel segments. Microangiectasias are inducible within 4 days of the onset of inflammation and lead to a greater than 10-fold local reduction in wall shear stress. These findings support the hypothesis that a preparatory step to lymphocyte transmigration involves structural adaptations in the inflammatory microcirculation. PMID- 12782791 TI - Oncogenic potential of TASK3 (Kcnk9) depends on K+ channel function. AB - TASK3 gene (Kcnk9) is amplified and overexpressed in several types of human carcinomas. In this report, we demonstrate that a point mutation (G95E) within the consensus K+ filter of TASK3 not only abolished TASK3 potassium channel activity but also abrogated its oncogenic functions, including proliferation in low serum, resistance to apoptosis, and promotion of tumor growth. Furthermore, we provide evidence that TASK3G95E is a dominant-negative mutation, because coexpression of the wild-type and the mutant TASK3 resulted in inhibition of K+ current of wild-type TASK3 and its tumorigenicity in nude mice. These results establish a direct link between the potassium channel activity of TASK3 and its oncogenic functions and imply that blockers for this potassium channel may have therapeutic potential for the treatment of cancers. PMID- 12782792 TI - Expression of prestin-homologous solute carrier (SLC26) in auditory organs of nonmammalian vertebrates and insects. AB - Prestin, the fifth member of the anion transporter family SLC26, is the outer hair cell molecular motor thought to be responsible for active mechanical amplification in the mammalian cochlea. Active amplification is present in a variety of other auditory systems, yet the prevailing view is that prestin is a motor molecule unique to mammalian ears. Here we identify prestin-related SLC26 proteins that are expressed in the auditory organs of nonmammalian vertebrates and insects. Sequence comparisons revealed the presence of SLC26 proteins in fish (Danio, GenBank accession no. AY278118, and Anguilla, GenBank accession no. BAC16761), mosquitoes (Anopheles, GenBank accession nos. EAA07232 and EAA07052), and flies (Drosophila, GenBank accession no. AAF49285). The fly and zebrafish homologues were cloned and, by using in situ hybridization, shown to be expressed in the auditory organs. In mosquitoes, in turn, the expression of prestin homologues was demonstrated for the auditory organ by using highly specific riboprobes against rat prestin. We conclude that prestin-related SLC26 proteins are widespread, possibly ancestral, constituents of auditory organs and are likely to serve salient roles in mammals and across taxa. PMID- 12782793 TI - When houseguests become parasites: sympatric speciation in ants. PMID- 12782794 TI - RNA polymerase II at initiation. PMID- 12782795 TI - Dengue virus envelope glycoprotein structure: new insight into its interactions during viral entry. PMID- 12782796 TI - Making virtual screening a reality. PMID- 12782797 TI - Voice, speech, and swallowing outcomes in laser-treated laryngeal cancer. AB - OBJECTIVE: To describe preliminary voice, speech, and swallowing outcomes in patients treated by endoscopic laser excision of laryngeal cancer with or without adjuvant radiation therapy. STUDY DESIGN: Retrospective review. METHODS: Seventeen surgically treated patients (five T2 glottic and 12 clinically staged T2 supraglottic squamous cell carcinomas) participated in the study. Self-ratings of voice (Voice Handicap Index) and swallowing (M. D. Anderson Dysphagia Inventory) were completed, as well as independent auditory-perceptual ratings of voice and speech recordings. RESULTS: Although no significant difference between Voice Handicap Index, M. D. Anderson Dysphagia Inventory, and listener ratings was identified based on tumor site and irradiation status, there was a trend toward poorer outcomes in patients who received adjuvant radiation therapy. Whereas the patients having supraglottic cancer tended to report better voice but poorer swallowing outcomes, the glottic cancer group displayed the opposite pattern. Severity on Voice Handicap Index correlated significantly with listener severity ratings of speech, suggesting that the patients' perception of their voice handicap was similar to the listeners' judgments of their speech severity. CONCLUSIONS: The results suggest the following trends: 1) Adjuvant radiation therapy was associated with poorer outcomes for voice, speech, and swallowing and may be associated with more impairment than surgery alone and 2) poorer outcomes on voice and swallowing were observed for the glottic and supraglottic cancer groups, respectively. To bolster these preliminary findings, additional outcomes studies in patients treated with conservation therapy are needed. PMID- 12782798 TI - Airway management after maxillectomy: routine tracheostomy is unnecessary. AB - OBJECTIVES/HYPOTHESIS: There is a paucity of data to guide the optimal management of the airway in patients after maxillectomy. The decision on whether a concomitant tracheostomy is needed is often dictated by the surgeon's training and experience. We reviewed our experience with maxillectomy to assess the need for tracheostomy in postoperative airway management. STUDY DESIGN: Retrospective analysis at a university hospital. METHODS: We identified 121 patients who underwent 130 maxillectomies between October 1990 and September 2001. Twenty-four of these were total (all six walls removed), 45 were subtotal (two or more walls removed), and 61 were limited (only one wall removed). Reconstruction ranged from none to microvascular free flap, with split-thickness skin graft being the most common reconstructive option. RESULTS: Only 10 tracheostomies (7.7%) were performed at the time of maxillectomy. These included four tracheostomies in patients who underwent bulky flap reconstruction, two tracheostomies in patients who underwent both flap reconstruction and mandibulectomy, one tracheostomy in a patient who underwent mandibulectomy, one tracheostomy in a patient with mucormycosis in anticipation of prolonged ventilatory support postoperatively, and two tracheostomies at the surgeons' discretion because of concern for upper airway edema. Among the 111 patients who underwent 120 maxillectomies without concomitant tracheostomy, 1 patient (0.9%), a 74 year-old man with oxygen dependent chronic obstructive pulmonary disease, required repeat intubation on day 3 and again on day 10 after the surgery, because of respiratory failure; fiberoptic examination confirmed the absence of upper airway compromise. CONCLUSIONS: The routine performance of tracheostomy in patients undergoing maxillectomy is unnecessary. Selective use of tracheostomy may be indicated in situations in which mandibulectomy or bulky flap reconstruction is performed or a concern for postoperative oropharyngeal airway obstruction because of edema or packing exists. PMID- 12782799 TI - Radiation following surgery for oral cancer: impact on local control. AB - OBJECTIVES: The use of postoperative radiation therapy (RT) is commonly used in the treatment of patients with high-risk squamous cell carcinoma of the head and neck. However, few data exist that quantitate the incremental benefit of RT when administered following surgery. The retrospective study was designed to measure the incremental benefit of adjuvant RT after surgery on control of the primary lesion when compared with patients undergoing surgical therapy alone for squamous cell carcinoma of the oral cavity. STUDY DESIGN: Retrospective chart. METHODS: Previously untreated patients with squamous cell carcinoma of the floor of mouth or oral tongue who were treated between 1974 and 1998 were eligible for study. A minimum follow-up of 2 years was required. Tumor site, stage, and RT data were correlated with local control. RESULTS: Patients with missing or incomplete data and those lost to follow-up or dead as a result of intercurrent disease with follow-up of less than 2 years were censored. A group of 211 patients who could be evaluated was available for the study. Radiation therapy was administered postoperatively to 58 patients, and 153 patients were treated with surgery alone. In the group treated with combined therapy, the average irradiation dose was 5850 cGy; this group included 35 patients who received more than and 16 who received less than 5700 cGy. CONCLUSIONS: The incremental benefit in terms of control of tumor at the primary site for patients receiving postoperative RT was between 0% and 7%. Surgery alone controlled 80% to 85% of primary tumors. The dose of postoperative RT did not correlate with local control. PMID- 12782800 TI - Inhibition of epidermal growth factor receptor signaling decreases p63 expression in head and neck squamous carcinoma cells. AB - OBJECTIVES/HYPOTHESIS: Both the epidermal growth factor receptor (EGFR) and the p53 homologue p63 are overexpressed in a significant number of cases of head and neck squamous cell carcinoma (HNSCC). Epidermal growth factor receptor and p63 both possess oncogenic properties, including the potential to increase cell proliferation and antagonize apoptosis. ZD1839 ("Iressa") is an adenosine triphosphate-competitive inhibitor specific to the EGFR tyrosine kinase currently under evaluation as a chemotherapeutic agent in HNSCC. The objective was to investigate whether p63 expression is decreased after treatment of HNSCC cells with ZD1839. Downregulation of p63 by ZD1839 would identify a potential molecular relationship between EGFR signaling and p63 and could provide insight into the mechanism of action of ZD1839. STUDY DESIGN: In vitro examination of p63 expression after ZD1839 treatment. METHODS: A human HNSCC cell line, SCC-012, was treated with varying doses of ZD1839. p63 protein and messenger RNA levels were analyzed by Western and Northern blot analyses. The effect of ZD1839 on SCC-012 cell cycle was analyzed by flow cytometric analysis. RESULTS: In SCC-012 cells there was a dose-dependent decrease in p63 protein and messenger RNA levels over the course of ZD1839 treatment. Levels of phosphorylated MAPK decreased and p27KIP-1 levels increased after ZD1839 treatment. ZD1839 treatment induced a twofold increase in G1-phase cells and a 3.5-fold decrease in S-phase cells consistent with growth arrest. CONCLUSION: ZD1839 downregulates p63 expression at the messenger RNA level, suggesting that p63 is a downstream target of EGFR signaling. PMID- 12782801 TI - Adenocarcinoma ex-pleomorphic adenoma of the lacrimal sac and nasolacrimal duct: a case report. AB - OBJECTIVES: To present the clinical presentation, workup, surgical approach, and pathological findings of the first case report of a patient with adenocarcinoma ex-pleomorphic adenoma of the lacrimal sac and nasolacrimal duct. STUDY DESIGN: Retrospective review of the records of a case of adenocarcinoma ex-pleomorphic adenoma of the lacrimal sac and nasolacrimal duct. METHODS: The clinical presentation, workup, surgical approach, and pathological findings were reviewed. RESULTS: A 51-year-old man presented with a 10-year history of recurrent epiphora of the right eye. At dacryocystorhinostomy a small lesion was visualized within the lumen of the lacrimal sac. A biopsy specimen was consistent with adenocarcinoma. En bloc resection was accomplished using a lateral rhinotomy and medial maxillectomy. The final specimen showed adenocarcinoma ex-pleomorphic adenoma. The patient was given postoperative radiation therapy. He was free of disease 16 months after treatment. CONCLUSIONS: Lacrimal sac tumors should be considered in the differential diagnosis of chronic epiphora. Management of nasolacrimal adenocarcinoma requires complete surgical resection. Radiation treatment in and of itself is not curative but may be useful as adjuvant therapy. Carcinoma ex-pleomorphic adenoma can develop in the lacrimal sac and nasolacrimal duct. PMID- 12782802 TI - Correlation between facial pain or headache and computed tomography in rhinosinusitis in Canadian and U.S. subjects. AB - OBJECTIVES: Objectives were 1) to determine whether a correlation exists between facial pain or headache and sinus disease severity by computed tomography (CT) scan in patients with rhinosinusitis and 2) to compare disease severity and pain perception in two geographically diverse North American patient populations. STUDY DESIGN: Prospective patient questionnaire before CT scan of the paranasal sinuses. METHODS: Patients with refractory rhinosinusitis were recruited at the University of Texas Medical Branch (Galveston, TX) and the University of Alberta (Edmonton, Alberta, Canada). Before CT scanning, patients completed a pain questionnaire. All scans were interpreted by one neuroradiologist and were scored using the Lund-McKay, Harvard, and Kennedy staging systems for rhinosinusitis. RESULTS: Fifty-one patients completed questionnaires (27 were Canadian). There was no correlation between pain severity and disease severity reflected by any of the three staging systems used (P >.05). The mean pain score for the U.S. patients was 7.3, and for Canadian patients, 5.2. The mean CT scores for U.S. versus Canadian patients were as follows: Lund-McKay, 2.6 versus 6.6; Harvard, 0.7 versus 1.0; and Kennedy, 1.4 versus 2.2. The Canadian patients had more severe disease on CT scan (Lund-McKay, P .05). On the other hand, the endolymphatic sac as measured on T2-weighted image tended to be smaller than the vestibular aqueduct (P <.05) and tended to be larger than the lumen of the endolymphatic sac (P <.0005). CONCLUSION: Both sequences can precisely depict the endolymphatic sac; however, the proton density-weighted image is a more appropriate indicator of the actual anatomical configuration of the endolymphatic sac with surrounding connective tissue and vestibular aqueduct. PMID- 12782815 TI - A murine model of cholesteatoma-induced bone resorption using autologous dermal implantation. AB - OBJECTIVE: To investigate a novel murine model for dermal implant-induced osteolysis analogous to bone resorption observed in middle ear cholesteatoma. STUDY DESIGN: Animal experiment. METHODS: We placed autologous dermal implants on the surface of mouse calvaria. The calvaria were examined at days 1, 3, 5, 7, and 14 after implantation by histological study and tartrate-resistant acid phosphatase immunohistochemical processing to detect osteoclasts. RESULTS: Dermal implants showed a significantly increased osteoclast density compared with control samples. The dermal implant tissue remained viable and produced a robust, localized inflammatory osteolytic response on the adjacent calvarial surface. Osteoclasts were predominantly found on the surface of the calvarium with the greatest osteoclast density seen at 5 to 7 days after implantation. CONCLUSION: The mouse model is expected to be a useful tool for investigating the pathogenesis of localized inflammatory bone resorption related to cholesteatoma. PMID- 12782816 TI - Reversible canal wall down mastoidectomy for acquired cholesteatomas: preliminary results. AB - OBJECTIVES: The educational objectives were to discuss a new technique to manage acquired cholesteatomas and to compare this with standard approaches used for cholesteatoma removal. The study objective was to determine the feasibility of using a new approach, the reversible canal wall down approach, in conjunction with bone cement to remove acquired cholesteatomas and, potentially, preserve the ossicular chain. STUDY DESIGN: Retrospective review of five patients who underwent resection of their acquired cholesteatomas using a technique that maximizes exposure, yet preserves the posterior canal wall and, potentially, the ossicular chain. METHODS: In the setting of a tertiary otological referral center, five consecutive patients with acquired cholesteatomas underwent reversible canal wall down surgery. Four patients were male (ages, 41, 53, 26, and 10 y), and one patient was a woman [age, 51 y]). A reversible canal wall down mastoidectomy approach was used in the patients. Main outcome measures were intraoperative findings with respect to cholesteatoma removal and postoperative clinical examination, including postoperative audiograms. RESULTS: Five patients had dry ears postoperatively. One patient initially had granulation tissue forming on the posterior ear canal. This eventually resolved. The patients who had a postoperative audiogram after cholesteatoma resection with ossicular chain preservation noted an improvement or stabilization in their hearing. CONCLUSIONS: Using the reversible canal wall down approach in conjunction with bone cement, one is able to obtain excellent exposure of the cholesteatoma and, potentially, preserve the ossicular chain. This approach may be particularly useful in revision cholesteatoma surgery or in cases of acquired cholesteatoma in which the surgeon may be able to preserve the ossicular chain. PMID- 12782817 TI - Endonasal laser dacryocystorhinostomy: its role in anticoagulated patients. AB - OBJECTIVE: To report the results of endoscopic laser-assisted dacryocystorhinostomy in anticoagulated patients. STUDY DESIGN: A retrospective study of 16 consecutive anticoagulated patients with distal nasolacrimal duct obstruction treated by endoscopic laser-assisted dacryocystorhinostomy. METHODS: A case note review was made of all patients treated with endoscopic laser assisted dacryocystorhinostomy who were taking coumadin in two centers between 1993 and 2000. The parameters of age, gender, indications for surgery, surgical findings, complications, and outcome were analyzed. The mean follow-up time was 14 months (range, 9-26 mo). RESULTS: Fifteen of the 16 patients who were treated had an eventual successful outcome, but 6 patients required revision surgery. The patient whose symptoms were not improved was shown to have functional epiphora. No patient had a problem with primary or secondary epistaxis, and no patient required admission. A major benefit was the lack of disruption of anticoagulant therapy. CONCLUSIONS: Endoscopic laser-assisted dacryocystorhinostomy is a safe, efficient technique for the relief of distal nasolacrimal duct obstruction in anticoagulated patients. Not only does it avoid any disruption to their anticoagulant therapy, but it also can be performed as an outpatient procedure. PMID- 12782818 TI - Increased expression of interleukin-9, interleukin-9 receptor, and the calcium activated chloride channel hCLCA1 in the upper airways of patients with cystic fibrosis. AB - OBJECTIVES/HYPOTHESIS: Mucus overproduction is commonly found in airway disease in patients with cystic fibrosis. Interleukin-9 (IL-9) has been shown to mediate airway hyper-responsiveness and mucus overproduction. Recently, the calcium activated chloride channel hCLCA1 has been described to be upregulated by IL-9 and has been thought to regulate the expression of soluble gel-forming mucins. We sought to examine the expression of IL-9, interleukin-9 receptor (IL-9R), and hCLCA1 in the upper airway of patients with cystic fibrosis in comparison to healthy control subjects and to demonstrate the relationship of IL-9, IL-9R, and hCLCA1 expression with mucus production. STUDY DESIGN: Prospective design. METHODS: Biopsy samples from nasal polyps of four patients with cystic fibrosis, nasal mucosa of six patients with cystic fibrosis, sinus mucosa of eight patients with cystic fibrosis, and nasal mucosa of six healthy control subjects were stained with periodic acid-Schiff (PAS) to identify mucus glycoconjugates. IL-9, IL-9R, and hCLCA1 expression was determined by immunocytochemical study. RESULTS: We demonstrated significant increases in IL-9, IL-9R, and hCLCA1 immunoreactivity in the mucosa of patients with cystic fibrosis compared with that found in control subjects (P <.05). There were no significant differences between the different locations (nasal polyps, nasal mucosa, and sinus mucosa) in the patient group (P >.05). We also observed a significant increase in the number of mucus producing cells in biopsy specimens from patients with cystic fibrosis in comparison to control subjects. A positive correlation was found between hCLCA1 positive cells and IL-9-positive cells (correlation coefficient [r] = 0.79, P <.05) or IL-9R-positive cells (r = 0.92, P <.05). Moreover, a positive correlation was also present between PAS-positive (mucus-producing) cells and hCLCA1-positive cells (r = 0.64, P <.05) or IL-9R-positive cells (r = 0.64, P <.05). CONCLUSIONS: Increased expression of IL-9 and IL-9R, as well as upregulation of hCLCA1, in mucus-overproducing epithelium of patients with cystic fibrosis supports the hypothesis that IL-9 contributes to mucus overproduction in cystic fibrosis. Expression of hCLCA1 may also be responsible, in part, for the overproduction of mucus. These preliminary findings suggest that hCLCA1 might be an interesting new therapeutic target to control mucus overproduction in airway disease in patients with cystic fibrosis. PMID- 12782819 TI - Ipsilateral full-thickness forearm skin graft for covering the radial forearm flap donor site. AB - OBJECTIVES/HYPOTHESIS: The technique of the ipsilateral full-thickness forearm skin graft for covering the defect of radial forearm free flap (RFFF) improves aesthetic impairment at the recipient and donor sites by split-thickness skin graft repair and omits the need to make an extraoperative site for harvesting the skin graft. However, in this technique, the RFFF is limited in size. In the present study, we considered a model of the forearm and calculated the possible size of the RFFF for using this technique. METHODS: The calculation was conducted under assumptions that the isosceles-triangle skin graft is elevated as its height is twice the RFFF length in the direction of the forearm axis and that the forearm skin defect can be primarily closed with a width shorter than one-fourth of the wrist circumference. The calculation revealed that this technique is feasible when the RFFF width, that is, the length vertical to the forearm axis, is shorter than half of the wrist circumference. We repaired the RFFF defect using this technique in 15 patients with head and neck cancer in whom the RFFF size conformed to the above-mentioned condition. RESULTS: When the RFFF width was shorter than half of the wrist circumference and the isosceles-triangle skin graft was elevated as its height was twice the RFFF length, the RFFF defect could be repaired using this technique in all 15 patients. CONCLUSION: The above mentioned condition (that the RFFF width is shorter than half of the wrist circumference) is useful for determining whether or not the technique of ipsilateral full-thickness forearm skin graft can be used for covering the RFFF defect. PMID- 12782820 TI - Nontraumatic atlantoaxial subluxation (Grisel syndrome): a rare complication of otolaryngological procedures. AB - OBJECTIVE: At the conclusion of the report, the readers should be able to recognize the rare complication of adenotonsillectomy of Grisel syndrome, discuss its pathogenesis, and provide early, effective treatment. STUDY DESIGN: Analysis of the clinical presentation, plain radiographs, computed tomography scans, and magnetic resonance imaging scans of a child with Grisel syndrome after adenotonsillectomy. METHODS: Retrospective study of the case record and a review of the literature regarding the pathogenesis, treatment modalities, outcome, and prognosis after early and delayed treatment of Grisel syndrome. RESULTS: Pathophysiology of atlantoaxial subluxation revolves around the periodontoid vascular plexus that drains the posterosuperior pharyngeal region. Infectious and inflammatory emboli cause synovial engorgement, weakening paraspinal ligaments. Radiological studies play an important role in diagnosis. Treatment consists of cervical immobilization, muscle relaxants, analgesics, and antibiotics. Full resolution is expected with early conservative management. Failure to recognize the syndrome can lead to catastrophic results. CONCLUSIONS: Recognition of Grisel syndrome in addition to other rare complications of adenotonsillectomy requires a high index of suspicion. Early intervention is the critical factor for a positive outcome. However, delayed diagnosis is common and can result in catastrophic consequences, including neurological deficits, cosmetic deformity and, in rare instances, paralysis and death in the short term. PMID- 12782821 TI - Detection of Epstein-Barr virus-derived latent membrane protein-1 gene in various head and neck cancers: is it specific for nasopharyngeal carcinoma? AB - OBJECTIVE: The object of the study was to determine the incidence of the presence of Epstein-Barr virus-derived latent membrane protein-1 (LMP-1) gene in various head and neck cancers by polymerase chain reaction method. STUDY DESIGN: Prospective study. METHODS: During a 5-year period, polymerase chain reaction was used to investigate the presence of LMP-1 gene in various head and neck cancers from different locations and histopathological types, noncancerous nasopharyngeal biopsy samples, and tonsillectomy specimens from patients with chronic hypertrophic tonsillitis. RESULTS: Of 202 patients enrolled in the study, 53 were diagnosed by pathological study with oropharyngeal carcinoma, 45 with nasopharyngeal carcinoma, 26 with oral cavity carcinoma, 26 with laryngohypopharyngeal carcinoma, 31 with nasopharyngeal lymphoid hyperplasia, and 21 with tonsils with lymphoid hyperplasia. After the application of polymerase chain reaction, the LMP-1 gene was not detected in any sample from oral cavity carcinoma, laryngohypopharyngeal carcinoma, or nasopharyngeal lymphoid hyperplasia or from tonsillectomy specimens but was detected in only one case of tonsillar carcinoma. On the contrary, the LMP-1 gene was detected in 43 (95.6%) of 45 samples from patients with nasopharyngeal carcinoma. The statistical analysis shows a significant association (P <.001) between the presence of LMP-1 gene and tumor localization in the nasopharynx. CONCLUSIONS: The study shows that the presence of LMP-1 gene detected by polymerase chain reaction in the tumor cell is only significantly associated with tumor located in the nasopharynx, implying that Epstein-Barr virus plays a trifling role in the tumorigenesis of carcinomas arising from other head and neck locations. The polymerase chain reaction method that was used is a potential tool for screening nasopharyngeal carcinoma. PMID- 12782822 TI - Hereditary paraganglioma due to the SDHD M1I mutation in a second Chinese family: a founder effect? AB - OBJECTIVES/HYPOTHESIS: Hereditary paraganglioma is a rare condition that is inherited in an autosomal-dominant fashion. Four distinct loci have been associated with hereditary paraganglioma, including the SDHD, SDHC, and SDHB genes and a locus at 11q13. The SDHD, SDHC, and SDHB genes code for subunits of succinate dehydrogenase, which forms part of the mitochondrial respiratory chain. SDHD mutations are widely distributed along the gene with no apparent hot spots, although a founder effect has been described in the Dutch population. METHODS: Following a prior report of the SDHD M1I mutation in an Australian Chinese family, a second Chinese family with the same mutation is reported. The proband developed bilateral head and neck paragangliomas at age 34 years and a functioning adrenal pheochromocytoma and two extra-adrenal abdominal paragangliomas 7 years later. His brother had unilateral head and neck paraganglioma at age 39 years. Given the multicentricity of the proband's tumor and the familial clustering of paragangliomas, a clinical diagnosis of hereditary paraganglioma was made, and the proband was tested for a mutation in the SDHD gene. RESULTS: The proband was found to be heterozygous for the SDHD MII mutation that removes the start codon, and his brother subsequently tested positive for the same mutation. The family is not related to the Australian Chinese family. CONCLUSION: The finding suggests the possibility of a founder effect in the Chinese population and warrants further investigation. PMID- 12782823 TI - Bile acid induces cyclo-oxygenase-2 expression in cultured human pharyngeal cells: a possible mechanism of carcinogenesis in the upper aerodigestive tract by laryngopharyngeal reflux. AB - OBJECTIVES/HYPOTHESIS: Laryngopharyngeal reflux is a common event in patients with head and neck cancer. Bile acid is known to be related to tumor formation in the esophagus through the overexpression of cyclo-oxygenase-2 (COX-2), an enzyme that produces prostanoids. To better understand the mechanism of the laryngopharyngeal reflux-cancer connection, we examined COX-2 expression by bile acid in cultured human pharyngeal mucosa cells. METHODS: COX-2 expression induction by various combinations of chenodeoxycholate and acidity was observed by Western blotting and reverse transcriptase-polymerase chain reaction. COX-2 promoter activity was also measured by luciferase promoter assay. RESULTS: Chenodeoxycholate, one of the bile acid components, was found to induce COX-2 expression in human pharyngeal cells. Moreover, the induction of COX-2 by chenodeoxycholate was enhanced by acidity in a dose-dependent manner, and the promoter activity of COX-2 was increased by chenodeoxycholate in SNU-1041, a human laryngeal cancer cell line, whereas the transcription of COX-2 was inhibited by actinomycin-D. CONCLUSION: Bile salts or acidic conditions, or both, can induce COX-2 expression in normal pharyngeal mucosa, which implies that laryngopharyngeal reflux has a role in the tumorigenesis of the upper aerodigestive tract. PMID- 12782824 TI - Results of orbital preservation for advanced malignant maxillary sinus tumors. AB - OBJECTIVE: The purpose of the study was to examine the oncological and functional outcomes of multimodality therapy for patients with advanced malignant maxillary sinus tumors that invaded the orbit. STUDY DESIGN: Retrospective study. METHODS: The medical records of 26 patients with orbital invasion were retrospectively analyzed. The patient group consisted of 16 men and 10 women, with a median age of 58 years. The mean follow-up period was 73 months. The most common disease was squamous cell carcinoma. Seven patients had nodal disease. All patients underwent simultaneous combined therapy consisting of conservative surgery through a sublabial incision, radiotherapy, and regional chemotherapy. Patients with nodal disease were treated with either irradiation or selective neck dissection. RESULTS: The 5- and 10-year overall survival rates were 68% and 51%, respectively. The 5- and 10-year local control rates were 66% and 51%, respectively. Overall survival rates and local control rates were significantly worse in patients with disease other than squamous cell carcinoma. Local control rates were significantly worse in patients with orbital apex disease than in patients without orbital apex disease. All 26 patients, despite orbital involvement, retained their orbital contents. Nineteen of these patients demonstrated adequate ocular function. CONCLUSIONS: Combined therapy with conservative surgery, radiotherapy, and regional chemotherapy is an effective method for local control and preservation of ocular function. However, performing orbital conservation procedure in patients with disease other than squamous cell carcinoma and with orbital apex disease must be considered carefully. PMID- 12782826 TI - A randomized controlled trial of early oral feeding in laryngectomized patients. AB - OBJECTIVE: To evaluate the safety and efficacy of early oral feeding by comparing it with feeding through primary tracheoesophageal puncture after total laryngectomy with primary pharyngeal closure. STUDY DESIGN: A prospective, randomized, controlled study. METHODS: Patients who underwent total laryngectomy with primary pharyngeal closure and who were candidates for primary voice restoration (an in whose cases primary tracheoesophageal puncture [TEP] was created) were included. After total laryngectomy, patients were randomly assigned to either the oral group (study group) or the TEP group (control group). Patients in the oral group were fed orally with a clear liquid diet on the first postoperative day, then advanced to a regular diet, whereas patients in the TEP group were fed through tracheoesophageal puncture and received nothing orally until the seventh postoperative day; then they were fed orally if fistula had not occurred. Standard criteria for discharge were used for all the patients. RESULTS: During a 3-year period, 67 patients were enrolled in the trial, and complete data were available for 65 patients (32 patients in the oral group, 33 patients in the TEP group). The two groups were similar for factors reported to influence the rate of pharyngocutaneous fistula. In three (9%) patients in the TEP group, fistula occurred on the 5th, 7th, and 14th postoperative days, respectively. Two (6.2%) fistulas occurred in the oral group on the sixth and eighth postoperative days, respectively. In patients without fistula, the mean length of hospital stay was 7.6 days (range, 4-19 d [SD = 3.1 d]) for the oral group and 8.2 days (range, 7-18 d [SD = 2.6 d) for the TEP group. There was no significant difference between two groups for either the incidence of fistula or the length of hospital stay. CONCLUSIONS: Initiation of oral feeding on the first postoperative day in patients undergoing total laryngectomy with primary pharyngeal closure is a safe clinical practice. However, it does not shorten the length of hospital stay for these patients. PMID- 12782825 TI - Primary and metastatic cancer of the parotid: comparison of clinical behavior in 232 cases. AB - OBJECTIVES/HYPOTHESIS: Parotid malignancy may develop as a primary cancer of salivary tissue or by metastatic involvement of parotid lymph nodes. The aim of the study was to compare the clinical behavior of primary and metastatic parotid cancers by analyzing patterns of treatment failure and clinical outcomes. STUDY DESIGN: Retrospective review of clinical and pathologic data prospectively accessioned onto a computerized database. METHODS: A prospectively documented series of 232 parotidectomies carried out for treatment of cancer from 1988 to 1999 was reviewed. There were 177 male and 55 female patients with a median age of 65 years (age range, 17-97 y). Median follow-up time was 4 years. Pathological groups included 54 patients with primary parotid cancer, 101 with metastatic cutaneous squamous cell carcinoma, 69 with metastatic melanoma, and 8 with other metastatic cancers. RESULTS: Neck nodes were clinically positive in 12 patients with primary cancer, 24 patients with squamous cell carcinoma, 16 with melanoma, and 2 with other metastatic malignancies. Conservative parotidectomy, preserving the main trunk of the facial nerve, was performed in 185 patients, and 47 patients had a radical parotidectomy sacrificing the facial nerve. There were 54 therapeutic and 110 elective neck dissections. Adjuvant radiotherapy was given to 39 patients with primary cancer, 86 with squamous cell carcinoma, 50 with melanoma, and 8 in the other metastatic group (78% of the patients in the series). Local control rates at 5 years in the four groups were 86%, 75%, 94%, and 100%, respectively (P <.01). Survival rates at 5 years were 77%, 65%, 46%, and 56%, respectively (P <.01). CONCLUSIONS: The pattern of parotid malignancy is unique in Australia because of the high incidence of skin cancer, which can metastasize to the parotid gland. Metastatic cutaneous malignancy predominates. The pattern of failure and outcome varied depending on histological findings. Local failure occurred most often in metastatic squamous cell carcinoma, whereas patients with melanoma had the highest incidence of distant spread. PMID- 12782827 TI - A case of paranasal sinus angiolipoma. AB - OBJECTIVE: To present the first documented case of an angiolipoma of the maxillary sinus, our treatment of the patient, and the 1-year follow-up results. At the conclusion of the report, the reader should be able to discuss the incidence, treatment options, and long-term prognosis of angiolipomas of the paranasal sinuses. STUDY DESIGN: A case study reporting on a single patient with the diagnosis of maxillary sinus angiolipoma. METHODS: Review of diagnostic studies, the operative technique, and the patient's chart for the 1-year period after treatment. RESULTS: The patient remains free of disease 1 year after definitive surgical treatment. CONCLUSIONS: Angiolipomas are exceedingly rare in this location. The accepted treatment for this disease in other regions of the body consists of wide local excision with close follow-up. The extent of the disease found in this patient required a medial maxillectomy for en bloc resection. We would advocate such an approach for other physicians who are faced with a patient with this rare diagnosis. PMID- 12782828 TI - Use of WuScope for exchange of endotracheal tube in a patient with difficult airway. AB - OBJECTIVE: To describe a method for the exchange of a defective endotracheal tube using the WuScope in patients with difficult airways who cannot tolerate interrupted ventilation. STUDY DESIGN: Case report. METHODS: Detailed description of proposed modality for the endotracheal tube exchange. RESULTS: Exchanging a defective endotracheal tube in patients with a "difficult airway" and compromised oxygenation can be a challenging task. Performing fiberoptic visualization with the WuScope and using a "double intubation" technique may be an acceptable method for endotracheal tube exchange in some clinical situations. CONCLUSION: By using our technique the exchange of an endotracheal tube in a patient with a difficult airway may be achieved without interrupting the ventilation. PMID- 12782829 TI - Office-based pulsed dye laser treatment for hemorrhagic telangiectasias and epistaxis. PMID- 12782830 TI - Hoarseness after cardiac surgery: possible contribution of low temperature to the recurrent nerve paralysis. PMID- 12782831 TI - Tracheoesophageal puncture using a KTP laser. PMID- 12782832 TI - Ear, nose and throat disorders in children with Down syndrome. PMID- 12782833 TI - Aphorisms of the hero. PMID- 12782834 TI - Estimated incidence of pseudoexfoliation syndrome and pseudoexfoliation glaucoma in Olmsted County, Minnesota. AB - PURPOSE: To determine the incidence of newly diagnosed pseudoexfoliation syndrome and pseudoexfoliation glaucoma in residents of Olmsted County, Minnesota from 1976 to 1991. MATERIALS AND METHODS: The database of the Rochester Epidemiology Project was used to identify all patients residing in Olmsted County who were newly diagnosed with pseudoexfoliation syndrome from 1976 to 1991. The criterion used to diagnose pseudoexfoliation syndrome was the presence of pseudoexfoliation material on 1 or more anterior segment structures. Criteria used to diagnose pseudoexfoliation glaucoma were diagnosis of pseudoexfoliation syndrome with evidence of glaucomatous changes or ocular hypertension receiving therapy. The overall age and sex-adjusted annual incidences of pseudoexfoliation syndrome and pseudoexfoliation glaucoma, adjusted to the 1990 US white population, were determined with differences across age and sex using Poisson regression. RESULTS: Pseudoexfoliation syndrome was diagnosed in 290 patients (mean age, 73 +/- 10 years). Two hundred twenty-one (76%) were female. The overall age and sex adjusted annual incidence of pseudoexfoliation syndrome was 25.9 per 100,000. The age-adjusted incidence was higher in females than in males (32.7 vs 16.9 per 100,000; P<0.001). Pseudoexfoliation syndrome diagnoses increased with age, from 2.8 per 100,000 in persons 40 to 49 years to 205.7 per 100,000 in persons > or =80 years (P<0.001). Thirty-five patients were excluded from pseudoexfoliation glaucoma diagnosis. Of the remaining 255 patients, 113 (44%) were diagnosed with pseudoexfoliation glaucoma during the study period (mean age, 76 +/- 10 years), 84 (74%) of whom were female. The overall age- and sex-adjusted annual incidence of pseudoexfoliation glaucoma was 9.9 per 100,000. The age-adjusted incidence was higher in females than in males (11.7 vs 7.2 per 100,000; P < 0.001). Pseudoexfoliation glaucoma was increasingly diagnosed with age, rising from 0.6 per 100,000 in persons 40 to 49 years to 114.3 per 100,000 in persons > or =80 years, (P<0.001). CONCLUSIONS: In a defined population, the estimated overall age and sex-adjusted annual incidence of pseudoexfoliation syndrome was 25.9 per 100,000 population, while the estimated overall age- and sex-adjusted annual incidence of pseudoexfoliation glaucoma was 9.9 per 100,000 population. The incidence of both diseases was higher in females and increased with advancing age. PMID- 12782835 TI - Angle-closure glaucoma in teenagers. AB - OBJECTIVE: To report the occurrence of angle-closure glaucoma in 2 teenagers. DESIGN: Observational case reports, review of literature. METHODS: Review of case histories, examinations, biometries, visual fields, and ultrasound biomicroscopy findings in 2 teenagers with angle-closure glaucoma. MAIN OUTCOME MEASURES: Intraocular pressure, gonioscopy, Humphrey 24-2 visual field (SITA Standard), and ultrasound biomicroscopy. RESULTS: The first case involved a 15-year-old white male who presented with an intraocular pressure of 60 mm Hg in the right eye and 24 mm Hg in the left eye and 360-degree appositional closure in both eyes. Ultrasound biomicroscopy revealed prominent bilateral ciliary pigment epithelial cysts pushing the iris anteriorly towards the angle. The second case involved a 14-year-old white male with a strong family history of primary angle-closure glaucoma. The patient had pupillary block and an intraocular pressure of 24 mm Hg in the right eye and 40 mm Hg in the left eye on routine eye examination. Gonioscopy and ultrasound biomicroscopy revealed appositional closure of the angle in all 4 quadrants bilaterally. CONCLUSION: Primary angle-closure glaucoma is uncommon in younger individuals. Therefore, the finding of angle-closure glaucoma in a young individual should alert the physician to the possibility of a secondary cause of angle closure, such as iris pigment epithelial cysts. In addition, special attention to family history is important as the configuration of an occludable anterior chamber angle may, in some instances, be inherited. PMID- 12782837 TI - Does smoking affect intraocular pressure? Findings from the Blue Mountains Eye Study. AB - PURPOSE: To assess the relationship between smoking and intraocular pressure. MATERIALS AND METHODS: The Blue Mountains Eye Study examined 3654 residents aged 49 years and older in an area west of Sydney, Australia from 1992 to 1994. A trained interviewer collected a detailed history of smoking. Intraocular pressure was measured using Goldmann applanation tonometry; as the correlation between right and left eyes was very high, only right-eye data are presented. Participants using glaucoma medications or who had evidence of previous cataract surgery were excluded. RESULTS: Current smokers (15.8% of participants) had slightly higher mean intraocular pressures (16.34 mm Hg) than nonsmokers (16.04 mm Hg). Intraocular pressure (in the right eye) was significantly associated with current smoking, after adjusting for age and sex (P = 0.03). This association remained unchanged after simultaneous adjustment for other variables associated with intraocular pressure, including blood pressure, diabetes, myopia, glaucoma, family history, and pseudoexfoliation (P = 0.02). CONCLUSIONS: This study identified a modest cross-sectional positive association between current smoking and intraocular pressure. PMID- 12782836 TI - Changes in intraocular pressure during prolonged (7-day) head-down tilt bedrest. AB - PURPOSE: To determine the change in intraocular pressure (IOP) due to postural changes in young healthy volunteers. MATERIALS AND METHODS: Intraocular pressure was measured using a calibrated Pulsair noncontact tonometer in both eyes of 25 female volunteers in a sitting position and after 1, 3, and 10 minutes in a supine position. In the second part of the experiment (a 7-day -6 degrees head down tilt [HDT]), IOP (at 8 am, 12 am and 6 pm) and corneal thickness (12 am) were monitored in 8 female volunteers before, during, and after the HDT period. Blood pressure, hematocrit, plasma volume and osmolality, and plasma catecholamines concentrations were also measured. RESULTS: Intraocular pressure was significantly higher in the supine position (16.1 +/- 3.6 mm Hg) than in the sitting position, with a mean pressure difference of 2.23 +/- 2.9 mm Hg after 1 minute, 0.9 +/- 3 mm Hg after 3 minutes, and 1.9 +/- 3.8 mm Hg after 10 minutes in a supine position (P < 0.001). During the period of HDT, IOP values decreased significantly on the fifth day (13.3 +/- 1.6 mm Hg, P = 0.03) and the seventh day (12.7 +/- 1.7 mm Hg, P = 0.02) when compared with IOP in the supine position (14.26 +/- 2 mm Hg). The corneal thickness increased significantly (P < 0.0001) at day 5 (549.25 +/- 48.7 microm) and day 7 (540.31 +/- 46.9 microm) compared with baseline (532.45 +/- 38.6 microm). Two days after the end of the HDT bedrest, the mean supine IOP significantly increased (14.1 +/- 1.8 mm Hg, P = 0.003) and corneal thickness was similar to that found at baseline. The mean decrease of IOP was positively correlated with that of the plasma volume (-10%, r = 0.61, P = 0.02) and negatively correlated with the mean rise of hematocrit (r = -0.5, P = 0.07), variables that are considered to be indirect measures of plasma dehydration. CONCLUSIONS: During a 7-day HDT bedrest experiment in healthy women, eyes seemed to compensate the moderate rise of IOP described between a sitting and a supine position, and exhibited a slight and progressive average decrease of 1.3 mm Hg. These physiological modifications could be related to an ocular dehydration or to systemic cardiovascular and hormonal variations during bedrest. PMID- 12782838 TI - The Italian version of the 25-item National Eye Institute Visual Function Questionnaire: translation, validity, and reliability. AB - OBJECTIVE: To test the Italian adaptation of the 25-item National Eye Institute Visual Function Questionnaire (NEI-VFQ 25) and to investigate its validity and reliability. METHODS: This prospective observational study enrolled nonhospitalized patients with 1 of 5 chronic eye diseases, and a reference sample of subjects without eye diseases. Eligible participants had to be cognitively able to respond to a health status interview and be affected by 1 of the following eye conditions: senile cataract, age-related macular degeneration, glaucoma, diabetic retinopathy, and cytomegalovirus retinitis or low vision. A control group was composed of subjects with no evidence of underlying eye diseases. The Italian self-administered versions of Medical Outcomes Study Short Form and of the NEI-VFQ 25 were administered to all participants. Reliability and validity of the Italian translation of the NEI-VFQ 25 were tested using statistical methods. RESULTS: Statistical analysis points out that the Italian version of the NEI-VFQ 25 has good validity, discriminatory power, internal consistency, and reliability. CONCLUSIONS: This Italian version of the NEI-VFQ 25 shows psychometric properties comparable to those of the American version, and thus can be used in clinical research as a specific measure of quality of life in patients with chronic eye diseases. PMID- 12782839 TI - Laterality of the performance of glaucoma mass screening using frequency-doubling technology. AB - PURPOSE: This study investigated laterality during the performance of glaucoma mass screening with a frequency-doubling technology perimetry test. MATERIALS AND METHODS: A frequency-doubling technology screening mode (C-20-1, version 2.6) test was performed on both eyes of 14,784 persons. Subjects with visual field abnormalities detected by the frequency-doubling technology test or with fixation error underwent retesting without a specified interval for rest. Consequently, 206 subjects who fulfilled the screening criteria of the frequency-doubling technology-based glaucoma screening protocol [FDT-GSP(+)] were further investigated using the Humphrey visual field analyzer (30-2). As a result, 74 right eyes and 57 left eyes were shown to have definite glaucoma. RESULTS: Frequency-doubling technology data for the left eye demonstrated a significantly (P<0.001) higher rate of artifacts, such as no reproducibility of results between the first and second tests (left/right: 2.4%/1.7%) as well as fixation errors (left/right: 2.8%/1.0%). The false-positive rate of the FDT-GSP for glaucoma was more than 1.5-fold higher in the left eye than in the right eye (16.3%/9.8%). In the case that either eye exhibited FDT-GSP(+), the positive predictive value of the FDT-GSP for definite glaucoma in the left eye was almost half of that in the right eye (28.4% vs. 53.8%). Specificity of the FDT-GSP for detection of definite glaucoma also exhibited a lower trend (P = 0.097) in the left eye (44.6%) than in the right eye (55.3%), but the sensitivity of the test was similar in both eyes (91.2% vs. 90.5%, respectively). CONCLUSIONS: When frequency-doubling technology based mass screening is performed on the general population, performance is lower for the left eye than for the right eye. This performance disparity is likely to be primarily associated with a difference in specificity. PMID- 12782840 TI - Optic disc hemorrhage in Asian glaucoma patients. AB - PURPOSE: To assess the prevalence of optic disc hemorrhage in Asian patients with established glaucoma using a recently described, highly sensitive detection method. MATERIALS AND METHODS: Patients entering a trial of glaucoma filtering surgery in Southeast Asia (The Singapore 5FU Study) were assessed for the presence or absence of optic disc hemorrhage using stereo flicker chronoscopy of stereophotograph pairs. RESULTS: A total of 167 patients (117 males and 50 females) were assessed, of whom 128 were Chinese. Primary open-angle glaucoma was diagnosed in 90 patients, primary angle-closure glaucoma in 69 patients, and pseudoexfoliative or pigment dispersion glaucoma in 7 patients. Five eyes of 5 patients had disc hemorrhage at enrollment (5/167), a rate of 2.99%. Four patients with disc hemorrhage had primary open-angle glaucoma and 1 had primary angle-closure glaucoma. There were no significant differences in global visual field indices, AGIS scores, or intraocular pressures between eyes with and without disc hemorrhage. All disc hemorrhages were seen in eyes prior to trabeculectomy. CONCLUSIONS: The prevalence of disc hemorrhage in the current study was comparable to that seen in primary open-angle glaucoma in clinic-based studies of white patients, but far less than that reported in normal-tension glaucoma studies. Hemorrhages were more common in primary open-angle glaucoma than primary angle-closure glaucoma. PMID- 12782841 TI - 24-hour monitoring of intraocular pressure in glaucoma management: a retrospective review. AB - PURPOSE: Diurnal variation in intraocular pressure (IOP) is well recognized, yet important decisions in glaucoma management are frequently made after 1 or 2 IOP measurements. Twenty-four-hour monitoring of IOP may identify IOP variation and spikes. This study determined the value of 24-hour IOP monitoring in routine clinical practice. METHODS: Data were acquired retrospectively from case notes of 29 glaucoma patients sequentially admitted for 24-hour IOP monitoring while taking their established antiglaucoma therapy. RESULTS: While there was no difference between the mean clinic (office) and mean 24-hour IOP measurements, the peak IOP during 24-hour monitoring was on average 4.9 mm Hg higher than the peak clinic IOP (P<0.0001). In 4 (13.8%) patients, the peak IOP over 24 hours was at least 12 mm Hg higher than the clinic peak. Peak IOP values occurred outside normal office hours in 51.7% of patients. Twenty-four-hour IOP monitoring resulted in a change of clinical management in 23 (79.3%) patients, including 13 (44.8%) who were offered trabeculectomy. CONCLUSION: Twenty-four-hour monitoring of IOP frequently led to a change of glaucoma management by identifying IOP fluctuations and spikes. High IOP and wide diurnal IOP variation are considered major risk factors for glaucoma progression, and standard clinic follow-up evaluations failed to identify these phenomena. PMID- 12782843 TI - Retinal arterial diameter changes in progressive and nonprogressive glaucoma. AB - PURPOSE: To determine if the degree of retinal arterial diameter change is different between patients with progressive and nonprogressive open-angle glaucoma. MATERIAL AND METHODS: In this prospective cohort study, 44 eyes of 44 open-angle glaucoma patients (mean age, 67.5 years; age range, 52-84 years; mean follow-up period, 4.9 years; follow-up range, 1.3-7.5 years) were included. The change in arterial diameter between the baseline and the most recent follow-up optic disc photograph was determined. The diameter of the 4 major arteries was measured at the optic disc margin and at the thinnest and broadest locations within 1 optic disc diameter from the disc margin. Patients were stratified into progressing and nonprogressing groups according to visual field and optic disc criteria. RESULTS: Visual field progression was observed in 13 (30%) patients and optic disc progression in 24 (55%) patients. On average, the arterial diameters at the edge of the optic disc decreased significantly by 2.37% (95% CI, -3.31% to -1.41%) per year of follow-up (P<0.001). No significant difference in generalized or focal arterial narrowing was observed between progressive and nonprogressive groups regardless of the criterion used (P>0.462). With this sample, the power to detect a 10% difference in arterial narrowing between the 2 groups was 66%. There was no relationship between the rates of visual field progression and arterial diameter change in the whole group (P = 0.171) or in groups segregated into progressing and nonprogressing patients (P>0.104). CONCLUSION: Arterial diameters decreased in both progressive and nonprogressive glaucoma. In this study, there was little evidence that arterial narrowing was more pronounced in progressive disease. PMID- 12782842 TI - Tonography demonstrates reduced facility of outflow of aqueous humor in myocilin mutation carriers. AB - PURPOSE: To demonstrate the effect in vivo of the myocilin gene mutation Thr377Met on outflow facility of aqueous humor, as measured by tonography. MATERIALS AND METHODS: Forty-two members of a pedigree known to carry the Thr377Met mutation were examined for glaucoma, evaluated with tonography, and screened for myocilin mutations. Tonography was used to calculate the coefficient of aqueous outflow facility (C), as well as the ratio of the resting intraocular pressure to C (P(0)/C). Subjects were reexamined for glaucoma 5 years after tonography. RESULTS: Seven subjects were excluded because of previous treatment known to alter facility of aqueous outflow. The mean outflow facility of the eyes of the 12 subjects carrying the Thr377Met mutation was significantly reduced compared with the 23 non-carriers' eyes using both C (P<0.001) and P(0)/C (P<0.001). Reduced outflow facility was also demonstrated in those mutation carriers who were not yet expressing clinical signs of glaucoma or ocular hypertension when measured using C (P = 0.015) and P(0)/C (P = 0.001). After 5 years, progression towards glaucoma had occurred in 5 of the myocilin mutation carriers, 2 of whom showed bilateral progression; 3 carriers remained completely normal. Four subjects had bilateral glaucoma at the outset and remained unchanged. The carriers' eyes that progressed towards glaucoma had reduced outflow facility compared with those that remained normal, although the difference was not statistically significant. CONCLUSIONS: Carriers of the myocilin Thr377Met mutation have reduced outflow facility, which may be detected prior to developing glaucoma. Tonography was not seen to be clinically useful in predicting progression towards glaucoma. PMID- 12782844 TI - Evaluation of retinal nerve fiber layer thickness in the area of apparently normal hemifield in glaucomatous eyes with optical coherence tomography. AB - PURPOSE: The authors investigated the correlation between visual field defects detected by automated perimetry and the thickness of the retinal nerve fiber layer measured with optical coherence tomography, and examined whether there is a decrease in retinal nerve fiber layer thickness in the apparently normal hemifield of glaucomatous eyes. PATIENTS AND METHODS: Forty-one patients with glaucoma and 41 normal control subjects were included in this study. Statistical correlations between the sum of the total deviation of 37 stimuli of each hemifield and the ratio of decrease in retinal nerve fiber layer thickness were evaluated. The statistical difference between the retinal nerve fiber layer thickness of the apparently normal hemifield in glaucomatous eyes and that of the corresponding hemifield in normal subjects was also evaluated. RESULTS: There was a statistically significant correlation in the sum of the total deviation and retinal nerve fiber layer thickness decrease ratio (superior hemifield, P = 0.001; inferior hemifield, P = 0.003). There was no significant decrease in retinal nerve fiber layer thickness in the area that corresponded to the normal visual field in the hemifield defect with respect to the horizontal meridian in glaucomatous eyes (superior side, P = 0.148; inferior side, P = 0.341). CONCLUSIONS: Optical coherence tomography was capable of demonstrating and measuring retinal nerve fiber layer abnormalities. No changes in the retinal nerve fiber layer thickness of the apparently normal hemifield were observed in glaucomatous eyes. PMID- 12782845 TI - Retinal nerve fiber loss pattern in high-tension glaucoma by optical coherence tomography. AB - PURPOSE: To identify the pattern of retinal nerve fiber layer thinning in high tension primary chronic open-angle glaucoma (HT-PCOAG) using optical coherence tomography. METHODS: Retinal nerve fiber layer thickness was assessed with optical coherence tomography in 68 healthy controls and 68 age- and refractive error-matched patients with HT-PCOAG were recruited. Patients were divided into 3 subgroups according to their visual field mean deviation: early (> -3 dB), moderate (-3 to -7 dB), and moderately advanced (-7 to -15 dB). RESULTS: Retinal nerve fiber layer thickness values in inferotemporal and superotemporal regions were significantly lower in all groups of glaucomatous eyes than in healthy eyes (P<0.001). Inferotemporal thickness values were significantly lower than the superotemporal thickness values (P<0.001) in the early glaucomatous stage, but not significantly so in moderate and moderately advanced stages. CONCLUSIONS: Besides a diffuse retinal nerve fiber layer defect, a localized defect may present in the inferotemporal region in early-stage HT-PCOAG. These results also suggest that optical coherence tomography shows promise in providing quantitative data about the location and extent of retinal nerve fiber layer injury in glaucoma, and the findings are consistent with previous knowledge. PMID- 12782847 TI - Morphologic classification of filtering blebs after glaucoma filtration surgery: the Indiana Bleb Appearance Grading Scale. AB - PURPOSE: To establish a new classification system for filtering blebs according to clinical morphologic parameters. The purpose of this classification system is to provide a uniform and objective assessment of bleb appearance and establish a framework system through which outcomes of filtration surgery may be better correlated to clinical morphology. MATERIALS AND METHODS: The Indiana Bleb Appearance Grading Scale contains a set of photographic standards illustrating a range of filtering bleb morphology selected from the slide library of the Glaucoma Service at the Indiana University Department of Ophthalmology. These standards consist of slit lamp images for grading bleb height, extent, vascularity, and leakage with the Seidel test. For grading, the morphologic appearance of the filtration bleb is assessed relative to the standard images for the 4 parameters and scored accordingly. Fifty-one clinical bleb photographs were evaluated and scored by 3 glaucoma subspecialists in a masked fashion according to the scale. RESULTS: For all of the grading scales, high interobserver agreement was found using the scale to classify the appearance of filtering blebs (height +0.76; extent +0.78; vascularity +0.90, interclass correlation coefficient for consistency using a 2-way mixed effect model). CONCLUSION: The Indiana Bleb Appearance Grading Scale is a simple, reproducible, yet comprehensive system for classifying the morphologic slit lamp appearance of filtration blebs. PMID- 12782848 TI - Bleb-limiting conjunctivoplasty for symptomatic circumferential trabeculectomy blebs. AB - PURPOSE: To describe a new surgical technique for limiting symptomatic circumferential conjunctival blebs, an uncommon complication after trabeculectomy. METHODS: Four eyes of 2 patients underwent conjunctivoplasty to limit the extent of symptomatic circumferential blebs. An 8/0 virgin silk corneal traction suture was used to allow better exposure of the conjunctiva. Radial conjunctival and Tenon incisions were made down to bare sclera in approximately the 10:30 and 1:30 clock hour positions. The conjunctival incisions were sutured, tacking down to the sclera. RESULTS: Immediate flattening of the interpalpebral bulbar conjunctiva was noted on the first postoperative day and there was no elevation of intraocular pressure or loss of bleb function. Both patients experienced a rapid improvement in their symptoms and no complications of the procedure were noted. Recurrence of bleb extension occurred in 1 eye 4 months postoperatively, and was treated with a repeat limiting conjunctivoplasty incision with a good result. CONCLUSIONS: Early results show that bleb-limiting conjunctivoplasty is an effective means of treating symptomatic circumferential trabeculectomy blebs. PMID- 12782846 TI - Optic disc morphometry correlated with confocal laser scanning Doppler flowmetry measurements in normal-pressure glaucoma. AB - PURPOSE: To examine the relationship between morphologic optic disc parameters and hemodynamic parameters as measured by confocal laser scanning Doppler flowmetry in patients with normal-pressure glaucoma. METHODS: The study included 91 eyes of 54 patients with normal-pressure glaucoma (mean age: 57.7 +/- 9.8 years), and 136 eyes of 77 age-adjusted normal controls. Color stereo optic disc photographs were morphometrically examined, and confocal laser scanning flowmetry (Heidelberg Retinal Flowmeter) in the neuroretinal rim inside of the optic disc, and in the retina close to the temporal and nasal border of the optic nerve head was performed. RESULTS: Mean confocal laser scanning flowmetric measurements in the neuroretinal rim, temporal parapapillary retina, and nasal parapapillary retina were significantly (P<0.03) lower in the normal-pressure glaucoma group than in the age-adjusted control group. Correspondingly, mean confocal laser scanning flowmetric measurements within the neuroretinal rim decreased significantly, with relatively low correlation coefficients, decreasing neuroretinal rim area (P = 0.016; correlation coefficient r2 = 0.026), and increasing mean visual field defect (P = 0.011; r2 = 0.029). Measurements were statistically independent of alpha zone (P = 0.38; r2 = 0.004) and beta zone (P = 0.57; r2 = 0.002) of parapapillary atrophy. CONCLUSIONS: Confocal laser scanning flowmetric measurements within the neuroretinal rim were lower in eyes with normal-pressure glaucoma than in age-matched normal eyes. Confocal laser scanning flowmetric measurements decrease with increasing glaucomatous optic nerve damage. There is, however, a marked variability preventing a clear relationship between stage of glaucoma and decrease in confocal laser scanning flowmetric measurements. The correlation between parapapillary atrophy and confocal laser scanning flowmetric measurements is not statistically significant in normal pressure glaucoma. PMID- 12782849 TI - Biocompatibility of the Ex-PRESS miniature glaucoma drainage implant. AB - PURPOSE: Based on lessons learned from earlier attempts, a novel miniature glaucoma implant, Ex-PRESS, was developed in 1998. The current study summarizes the histopathologic evaluation of this device implanted in the eyes of rabbits. METHODS: The device was implanted into the anterior chamber at the corneoscleral junction in 1 eye each of 8 white New Zealand rabbits, while the contralateral eye served as control. Three and 6 months after implantation, the rabbits were killed and their eyes were enucleated and processed histologically, leaving the device in situ when sectioning. RESULTS: Three and 6 months postoperatively, the local tissue reaction typically consisted of an enveloping, thin, mature, fibrotic capsule (thickness <0.04 mm), devoid of inflammatory cells. This capsule surrounded approximately 25% of the implant surface area present in the sections. The lumina of the devices were devoid of inflammatory exudates or other obstructions in all specimens examined, suggesting free flow of fluid. CONCLUSIONS: The implantation of the Ex-PRESS miniature glaucoma shunt resulted in minimal capsular reaction. Considering the high reactivity of the rabbit eye, it is possible that this implant will induce a smaller cellular inflammatory reaction in the human eye. PMID- 12782850 TI - Bleb dysesthesia. PMID- 12782851 TI - Ophthalmodynamometry in eyes with dilated episcleral veins. AB - PURPOSE: To determine the central retinal vessel collapse pressure in patients with dilated episcleral veins. METHODS: This clinical noninterventional comparative study included a study group of 10 eyes (6 patients) with dilated episcleral veins, and a control group consisting of 50 eyes of 40 patients with cataract or refractive problems. With topical anesthesia, a Goldmann contact lens fitted with a pressure sensor in its holding ring was placed onto the cornea. Pressure was asserted onto the globe by pressing the contact lens, and the pressure value at the time when the central retinal artery and vein started pulsating was noted as central retinal artery and vein collapse pressure. RESULTS: Central retinal vein collapse pressure was significantly higher (P<0.001) in the study group with dilated episcleral veins than in the control group (37.9 +/- 33.8 vs 5.1 +/- 8.4 relative units [RU]). In the central retinal artery, there was not significant difference in diastolic collapse pressure measured between the study and control groups (78.2 +/- 22.8 vs 74.2 +/- 18.8 RU, respectively; P = 0.74). CONCLUSIONS: As measured by a new ophthalmodynamometer with biomicroscopic visualization of the central retinal vessels during the examination, the central retinal vein collapse pressure measurements were significantly higher in eyes with dilated episcleral veins than in control eyes. Ophthalmodynamometric estimation of the central retinal vein collapse pressure may be helpful in the assessment of patients with dilated episcleral veins. PMID- 12782852 TI - Digital infrared photography assists in the detection of iris and ciliary body cysts. AB - PURPOSE: To report a case that illustrates the utility of digital infrared photography in the detection of iris and ciliary body cysts in an asymptomatic patient. MATERIALS AND METHODS: Digital infrared photography of the iris was performed in both eyes of a 50-year-old white woman who presented with bilateral endothelial pigment dusting and subtle iris transillumination defects. RESULTS: Infrared photography revealed a pronounced, rounded patch of transillumination in the periphery of the left iris. Also observed were focal, rounded regions of reduced transmittance in the peripheral iris of both eyes. These findings suggested the presence of bilateral iris and ciliary body cysts. High-resolution ultrasound biomicroscopic images were consistent with this suspicion. CONCLUSIONS: Digital infrared photography of the iris may be useful in the detection and evaluation of iris and ciliary body cysts. PMID- 12782854 TI - On the usefulness of the parts of the body. PMID- 12782855 TI - Biomechanical evaluation of posterior screw fixation in cadaveric cervical spines. AB - Sixteen fresh-frozen spines from cadavers (C4-T1) were randomized on the basis of dual energy xray absorptiometry analysis of bone mineral density. The specimens were subjected to physiologic loads ( 10 years) after cervical laminoplasty was assessed and the postoperative problems were clarified. One hundred thirty-three patients had laminoplasty between 1981 and 1989 for treatment of cervical myelopathy and 126 patients were available for the current study. The clinical results were evaluated using the Japanese Orthopaedic Association score. The radiologic findings were analyzed by postural anomalies and range of motion. The average preoperative score was 9.1 points, and the postoperative score improved to 13.7 points within a year. The Japanese Orthopaedic Association score and recovery rate were maintained at 13.4 points and 55.1% at the last followup. In 20 patients, the Japanese Orthopaedic Association score worsened during the followup. The causes of deterioration were axial spread of ossification of the posterior longitudinal ligament, other spinal lesions, cerebral infarction, and peripheral neuropathy. Postoperative cervical radiculopathy occurred in nine patients. Postoperative radiculopathy resolved in five patients, but remained in four patients. Kyphotic changes were observed in eight patients. The recovery rate in patients with kyphosis was poor. The postoperative range of motion decreased to 25.1% of preoperative range of motion. Sixty one percent of patients had a reduction of range of motion. Satisfactory results of cervical laminoplasty were maintained for more than 10 years after surgery; however, there were several postoperative problems, such as neurologic deterioration, postoperative radiculopathy, progression of kyphosis, and range of motion limitation. PMID- 12782869 TI - Measurements of pelvic flexion angle using three-dimensional computed tomography. AB - The purpose of the current study was to evaluate whether safe acetabular component position depends on differences in pelvic location between the supine, standing, and sitting positions. The subjects of the current study were 101 patients who had total hip arthroplasty. Anteroposterior radiographs of the pelvis with the patients in the supine, standing, and sitting positions were obtained preoperatively and 1 year after total hip arthroplasty. Computed tomography images of the pelvis were obtained preoperatively. Using image matching between the three-dimensional computed tomography model and anteroposterior radiograph, pelvic flexion angles with the patient in the supine, standing, and sitting positions were calculated. The mean preoperative pelvic flexion angle was 5 degrees +/- 9 degrees (range, -37 degrees -30 degrees ) in the supine position, 3 degrees +/- 12 degrees (range, -46 degrees -33 degrees ) in the standing position, and -29 degrees +/- 12 degrees (range, -62 degrees -10 degrees ) in the sitting position. Because there was much intersubject variability in pelvic flexion angle, it is not appropriate to determine orientation of the acetabular component from anatomic landmarks. In 90% of the cases, the difference in pelvic flexion angle between the supine and standing positions preoperatively was 10 degrees or less. In 90% of the cases, there was 20 degrees or greater extension of the pelvis from the supine position to the sitting position preoperatively, and the safe range of flexion of the hip from anterior prosthetic impingement in the sitting position was 20 degrees or greater than that in the supine position. Preoperative pelvic position in each case was almost completely maintained 1 year after total hip arthroplasty. It is reasonable to regard the pelvic position in the supine position as the functional pelvic position and proper pelvic reference frame in determining optimal orientation of the acetabular component in 90% of cases before and 1 year after total hip arthroplasty, although an adjustment of orientation of the acetabular component was needed for the remaining cases. PMID- 12782870 TI - Proximal kyphosis after short posterior fusion for thoracolumbar scoliosis. AB - Thoracolumbar idiopathic scoliosis usually is treated by anterior spinal fusion. However, short posterior spinal fusion that includes only the structural curve has been tried in a limited number of patients. The fusion may end cranially in the lower thoracic region and cause an increase in sagittal decompensation at the proximal junction. From July 1989 to July 1998, 14 patients were treated with thoracolumbar idiopathic scoliosis by short posterior spinal fusion. The lateral radiographs were evaluated preoperatively, immediately postoperative, and during followup. The focal kyphotic angle was used to examine the changes in focal sagittal alignment. A 10 degrees progression was defined as the radiographic criterion for the development of junctional kyphosis. Proximal junctional kyphosis occurred in six of the 14 patients, in which one patient needed revision surgery. In all six patients, the average preoperative lumbar lordosis was greater than 35 degrees, and decreased more than 10 degrees during surgery. In the five patients with a focal kyphotic angle larger than 10 degrees, four had proximal junctional kyphosis develop. According to the current findings, short posterior spinal fusion can be done only if the focal kyphotic angle proximal to the fusion is less than 10 degrees, and the lumbar lordosis must be preserved carefully during surgery. PMID- 12782871 TI - Efficacy of serotonin receptor blocker for symptomatic lumbar disc herniation. AB - Serotonin is one of the chemical mediators associated with nerve root inflammation and sciatic symptoms in lumbar disc herniation. The efficacy of serotonin 5-HT(2A) receptor blocker was examined in 44 patients with symptomatic lumbar disc herniation. A selective 5-HT(2A) receptor blocker (sarpogrelate hydroxychloride) was administered orally at a dose of 300 mg per day for 2 weeks. Visual analog scales of low back pain, sciatic pain, and numbness were significantly improved after the administration of the serotonin 5-HT(2A) receptor blocker. Clinical results were good (> 50% pain relief) in 23 patients, fair (25%-50% pain relief) in five patients, and poor (< 25% of pain relief) in 16 patients. Nineteen patients eventually required surgery because of muscle weakness or cauda equina symptoms. The effect of 5-HT(2A) blocker was good in 64% of patients who had uncontained disc herniation, whereas all patients with contained disc herniation had fair or poor results. Patients with uncontained disc herniation responded more favorably to the 5-HT(2A) blocker treatment than patients with contained disc herniation. A 5-HT(2A) blocker has the potential to block the cascade of acute nerve root inflammation and to alleviate symptoms in lumbar disc herniation. PMID- 12782872 TI - Categorization diminishes the reliability of hip scores. AB - Scoring systems frequently are used to assess the outcome of total hip arthroplasty. The result may be presented as a numeric value, or in descriptive terms such as excellent, good, fair, and poor (category system). The current study was done to investigate the influence of descriptive and numeric outcomes for interobserver reliability and interscore correlation of five different hip scores. Sixty-four patients (83 hips) were included in the study. The average age of the patients at followup was 70 years (range, 48-88 years). The average followup was 6.2 years (range, 2-17 years). For the numeric outcome a higher interobserver reliability (correlation coefficient, 0.71-0.81) and interscore correlation (correlation coefficient, 0.81-0.92) were found compared with the category system (interobserver reliability[correlation coefficient, 0.57-0.72]; interscore correlation [correlation coefficient, 0.46-0.62]). Findings from the study suggest that categorization of the results of total hip arthroplasty reduces interobserver reliability and interscore correlation. PMID- 12782873 TI - Arthroscopic treatment of a symptomatic hip plica. AB - Synovial plicae are detected most readily by diagnostic arthroscopy. As arthroscopic techniques are applied more frequently to various joints, it should be expected that the diagnosis of plica syndrome will become more common. The authors present a case of symptomatic plica in the hip of a competitive runner, who was treated with arthroscopic resection. PMID- 12782874 TI - Operative treatment of flatfoot with talocalcaneal coalition. AB - Rigid flatfoot or peroneal spastic flatfoot often is associated with a congenital fibrous, cartilaginous, or osseous union of two tarsal bones or more, potentially causing great difficulties in its treatment. Since 1996, 12 patients (14 feet) with painful flatfoot and restricted motion of the hindfoot attributable to talocalcaneal coalition, were treated by resection of the coalition and subtalar arthroereisis by a bioreabsorbable implant. The results were evaluated by the ankle hindfoot clinical rating system of the American Orthopaedic Foot and Ankle Society. These clinical results also were evaluated statistically. Radiographs and computed tomography scans were taken preoperatively and postoperatively. Eight (57.1% of patients) had excellent results, three (21.4% of patients) had good results, and three (21.4% of patients) had fair results. No poor results, or subjective or objective complications were reported. On the basis of these early results, arthroereisis by implanting a bioreabsorbable device after removal of the tarsal coalition, seems to be an effective procedure for the treatment of symptomatic flatfoot associated with talocalcaneal coalitions: correcting the relationship between talus and calcaneus, restoring the alignment of the hindfoot, and reducing pain. PMID- 12782875 TI - Late recurrence of clubfoot deformity: a 45-year followup. AB - Idiopathic clubfoot is one of the most common congenital deformities. Regardless of the mode of treatment, clubfoot has a tendency to relapse until the age of 5 years. Relapses are rare in patients after 5 years of age. A 45-year followup of a patient with idiopathic clubfoot treated as a newborn with the Ponseti method is reported. The patient had both clubfeet well-corrected as seen clinically and radiographically with this method. However, recurrent deformities developed bilaterally when the patient was 8 years old. Because of the late recurrence of this deformity, the patient had a thorough neurologic evaluation, which was normal. Treating physicians should be aware of the possibility of late recurrence in patients who have had complete correction of their clubfoot deformity. PMID- 12782876 TI - Secondary chondrosarcoma in osteochondroma: report of 107 patients. AB - Secondary chondrosarcomas are rare; recognition and diagnosis are difficult. Slow growth and late recurrence require long-term followup to understand the clinical course. In the current study, 107 patients had secondary chondrosarcoma arising in a solitary osteochondroma (61 patients) or multiple exostoses (46 patients). All histologic slides were reviewed without knowledge of the outcome, and radiologic studies were available for review in 71 cases. Patients with secondary chondrosarcoma were one to two decades younger than those with primary chondrosarcoma. Male preponderance and a predilection for flat bones were observed. The radiologic signs of sarcomatous degeneration included irregularity of the margin, inhomogeneous mineralization, and an associated soft tissue mass. The tumors generally were well-differentiated. Only 10 tumors were classified as Grade 2. Five-year and 10-year local recurrence rates were 15.9% and 17.5%, respectively, and 5- and 10-year mortality rates were 1.6% and 4.8% for patients having initial treatment at the authors' institution. Metastasis developed in five patients: in the lung in four patients and in the groin region in one patient. Most patients who died of tumor died of local recurrence. Wide excision had the lowest local recurrence rate. With successful surgical treatment, patients may have long-term disease-free survival. PMID- 12782877 TI - Radiation therapy for giant cell tumors of bone. AB - For giant cell tumors of bone, does radiotherapy provide a safe and effective treatment? This retrospective review includes 24 patients with 26 histologically diagnosed tumors treated with megavoltage radiotherapy between March 1972 and July 1996. Of the 10 recurrent tumors, five had an intralesional resection, two had a biopsy, and three had no biopsy before radiotherapy. Of the 16 previously untreated tumors, one was irradiated after a marginal resection, five after an intracapsular resection, and 10 after biopsy alone. The total doses ranged from 35 to 55 Gy (median, 43 Gy) in fractions of 1.67 to 2.33 Gy per day. Twenty of 26 tumors (77%) were controlled locally. All of the local recurrences occurred within the irradiated field. Five of six patients with local recurrence were treated successfully with additional surgery. Salvage surgery after local recurrence required amputation of an extremity in three patients and a total knee replacement in one patient. The ultimate local control rate was 96% with one patient alive with progressive disease. Lung metastases in one patient were treated successfully with surgery, chemotherapy, and radiotherapy. In one patient a radiation-induced sarcoma developed 22 years after treatment. The authors conclude that radiation therapy is a safe and effective treatment option for benign giant cell tumors of bone. A total dose greater than 40 Gy is the only variable found to significantly influence local control. PMID- 12782878 TI - Validation of a functional evaluation system in patients with musculoskeletal tumors. AB - The validity and reliability of the Musculoskeletal Tumor Society functional evaluation system were evaluated using the Nottingham Health Profile, the Short Form-36, and the EuroQol protocol to measure the quality of life of patients with malignant musculoskeletal tumors. Forty-nine patients were assessed and osteosarcoma around the knee was the most common type of tumor. Prosthetic reconstructions had been done in 55.1% of patients. In a content validity analysis, neither social nor psychologic domain was identified in the Musculoskeletal Tumor Society system. All items of the lower extremity were related to the same function-walking ability. The Musculoskeletal Tumor Society system had a strong correlation with other quality of life measures in the construct validity and the reliability, but each domain of the system was so comprehensive that it could not represent the quality of life properly. Furthermore, the preoperative and postoperative status of patients could not be compared using this system. The overall validity and reliability of the Musculoskeletal Tumor Society system seemed to be acceptable. However, this system may require additional development of the appropriate domains for evaluating the quality of life in patients with musculoskeletal tumors. PMID- 12782879 TI - Metachronous osteosarcoma: a report of five cases. AB - Five patients with primary osteosarcoma who were treated effectively with chemotherapy and resective surgery and in whom a metachronous tumor subsequently developed in another site but who never had evidence of pulmonary metastases are described. The original bone scans of the five patients showed only the primary site as being involved. After diagnosis and treatment of the initial tumor, at intervals ranging from 12 to 78 months (average, 39 months) another osteosarcoma developed in the patients at a distant bony site. After diagnosis and treatment of the second osteosarcoma, followup of these patients for a mean of 77 months (range, 24-96 months) after the appearance of the metachronous tumor and for a mean of 118 months (range, 99-150 months) after the diagnosis of the primary lesion showed that none had pulmonary metastases develop. No recurrences of the local tumors have occurred and four of the patients currently are alive and free of disease. One of the patients had a fatal acute myelogenous leukemia develop 144 months after the discovery of the primary osteosarcoma and 86 months after the appearance of the metachronous disease. At the time of her death, however she had no evidence of osteosarcoma in any site. PMID- 12782880 TI - Solitary fibrous tumor in the extremity: case report and review of the literature. AB - A solitary fibrous tumor is a relatively unusual neoplasm first described as a distinctive tumor arising from pleura. Some reports have shown that solitary fibrous tumors also affect extrathoracic regions. The current study presents a literature review with four additional patients with solitary fibrous tumor arising from the extremities to clarify clinicopathologic features. The current four patients were two males and two females, ranging from 17 to 60 years of age. Magnetic resonance imaging scans of the current patients showed inhomogeneous low to intermediate intensity signal on T1-weighted images and inhomogeneous intermediate to high intensity signal on T2-weighted images. Histologically, the tumors were composed of a haphazard proliferation of spindle cells, although cellularity was variable in each case. Two of the four tumors showed hypercellularity of spindle cells with focally myxomatous or hyaline changes, whereas myxomatous patterns with scattered spindle cells throughout the specimens were observed in the other two tumors. Immunohistochemically, all four patients showed positive immunoreactivity for CD34, and two tumors showed focally positive immunoreactivity for bcl-2 protein. During the followup of 12 to 54 months, neither local recurrence nor distant metastasis was detected after wide resection. Examination of the literature and the current patients suggests that solitary fibrous tumors in the extremities are likely to have a malignant potential, although most patients have a benign clinical course. Local wide resection and careful long-term followup are necessary for patients with solitary fibrous tumor in the extremities. PMID- 12782881 TI - Treatment of nonunions and osseous defects with bone graft and calcium sulfate. AB - The treatment of long bone nonunions and fractures with osseous defects is challenging. The results of 26 patients with either a persistent long bone nonunion or an osseous defect after an open fracture were reviewed. Each patient was treated with debridement of devitalized tissue, open reduction and internal fixation, and bone grafting using a mixture of autogenous iliac crest bone graft and medical grade calcium sulfate. The current study evaluated the union rate and associated complications for treatment of these injuries using this protocol. Each nonunion was confirmed intraoperatively, and healing was determined clinically by the patients' return to full activities without pain and radiographically by the presence of bridging trabeculae. Complications included persistent nonunion (four patients), wound drainage (five patients), wound drainage and cellulitis (one patient) and cellulitis alone (one patient). Using this treatment protocol, 22 patients (85%) achieved healing after one surgery and an additional two patients (92%) achieved healing after a second surgery. Medical grade calcium sulfate increases the volume of graft material, facilitates bone formation, and is safe in the treatment of nonunions and fractures with osseous defects. PMID- 12782882 TI - The effect of pronation and supination on the minimally displaced scaphoid fracture. AB - The amount of rotation that occurs at the scaphoid waist fracture site with pronation and supination of the forearm is studied in 10 upper extremities from cadavers. Two colinear metal markers were placed in the osteotomized scaphoid and a below-the-elbow cast was applied. Spiral volumetric computed tomography scanning of the scaphoid was done with multiplanar reformation to evaluate displacement of the metal markers. Four of the 10 specimens also were studied without any immobilization. The total magnitude of motion from pronation to supination averaged 0.2 mm in the specimens with a below-the-elbow thumb spica cast, and 2.4 mm in specimens without immobilization. The current study showed no significant rotation at the minimally displaced scaphoid waist fracture site during pronation and supination in a below-the-elbow cast. Furthermore, there is unacceptable rotation at the fracture site in the absence of a cast. Based on this study, a below-the-elbow thumb spica cast seems adequate for fracture immobilization; however, clinical correlation is needed. PMID- 12782883 TI - Open exchange locked nailing in humeral nonunions after intramedullary nailing. AB - Humeral nonunions after cannulated intramedullary nailing have been difficult to reconstruct. In the current study, 23 consecutive patients were treated by open exchange locked nailing with bone grafting. There were 16 men and seven women with a mean age of 46.2 years. The nonunions followed humeral locked nailing in eight patients, Seidel nailing in 13, and Kuntscher nailing in two. The average nonunion time was 14.7 months. The nonunions, located at the proximal (1/3) in four humeri, middle (1/3) in 15, and distal (1/3) in four, were antegrade nailed in 10 and retrograde nailed in 13. Nineteen had 8-mm nails and four had 7-mm nails. Supplementary wiring was used in 19 patients. The average followup was 21.4 months. With one surgery, all but one patient (96%) achieved osseous union in, on average, 16.3 weeks. One patient with chronic renal dialysis had persistent nonunion and an osteolytic supracondylar fracture. Other complications included one postoperative radial nerve palsy, one brachial artery injury, and one wire infection. At followup, all patients with solid union had excellent or satisfactory recovery of shoulder function. The average postoperative Neer score (90.7) was significantly better than the average preoperative score (68.5). Two patients had losses of elbow motion of 10 degrees and 20 degrees, respectively. This study shows that humeral nonunion after cannulated intramedullary nailing can be treated effectively by open exchange locked nailing with bone grafting. Supplementary wiring can compress the nonunion and facilitate bone healing. PMID- 12782884 TI - Placement of half-pins for supra-acetabular external fixation: an anatomic study. AB - An alternative location for placement of half-pins during pelvic external fixation is the dense supra-acetabular bone in the region of the anterior inferior iliac spine. Although these fixators have gained popularity, to the authors' knowledge there are no studies evaluating the potential anatomic risks of placement of half-pins in this area; no safe corridors have been defined. Additionally, pins are placed near the hip capsule and no studies exist defining the superior extent of the hip capsule which potentially may be violated by placing half-pins in this location. The purposes of the current study were to evaluate the neurovascular risks and accuracy of fluoroscopically guided percutaneous placement of supra-acetabular half-pins, and to evaluate the anatomic superior extent of the hip capsule. Ten fresh frozen cadaveric pelves were used. A 5-mm half-pin was placed in the supra-acetabular bone under fluoroscopic guidance. Iliofemoral dissection was done and the proximity of the half-pin to local neurovascular risks was measured with a caliper. The hip capsule was exposed and the superior extent of the hip capsule was measured. Intraosseous pin placement was evaluated by direct observation. Nine pins were completely in bone, one had partially exited posteriorly and laterally. The lateral femoral cutaneous nerve was at risk with a mean distance of 10 mm (range, 2-25 mm) from the half-pins. The femoral nerve and femoral artery were not at risk. The average superior extent of the hip capsule was 16 mm above the joint (range, 11-20 mm). Half-pins can be placed accurately and safely in the supra acetabular region using percutaneous techniques, appropriate soft tissue sleeves, and fluoroscopic guidance. Insertion of pins at least 2 cm above the hip is recommended to avoid potential hip capsule penetration. PMID- 12782885 TI - Vibrio vulnificus infection of the lower limb after fish spine injuries. AB - Five patients with soft tissue infection of the lower extremity caused by Vibrio vulnificus after penetrating injuries by fish spines are described. Despite previously reported dismal rates of morbidity and mortality associated with Vibrio septicemia, early measures including wide-spectrum intravenous antibiotics and surgical excision of devitalized tissues resulted in complete resolution in all cases. Although penetrating injury is the common route of infection, indirect seeding may occur by the contamination of open wounds or injury to a limb previously submerged in contaminated water. PMID- 12782886 TI - Treatment of osteomyelitis with local antibiotics delivered via bioabsorbable polymer. AB - The purpose of the current study was to show the efficacy and safety of an absorbable polymer (polycaprolactone) as an antibiotic delivery vehicle for treatment of osteomyelitis. An intramedullary osteomyelitis was induced in the femur of adult rabbits by Staphylococcus aureus inoculation after use of a sclerosing agent, and then treatment was done with intramedullary irrigation and implantation of a rod made of polycaprolactone, polycaprolactone plus 6% tobramycin, or polymethylmethacrylate plus 6% tobramycin. A control group received irrigation only. At defined intervals, the animals were euthanized and culture of the inoculated site was done. In addition, histologic sections of body tissues were made to look for signs of systemic toxicity of the implant. After 4 weeks of treatment, a statistically significant difference was found between the animals that were treated with irrigation alone and the animals that were treated with antibiotic-laden rods of polycaprolactone or polymethylmethacrylate. There was no difference between the antibiotic rod types. No histologic evidence of toxicity was found. Bioabsorbable rods of polycaprolactone are a safe and effective means of antibiotic delivery for treatment of osteomyelitis. PMID- 12782887 TI - The trochlea is bilinear and oriented medially. AB - Malfunctioning of total knee replacements often is related to patellofemoral problems. Because the trochlea guides the patella during flexion and extension, its geometry has a major influence in patellofemoral problems. There is controversy in the literature: relative to the mechanical axis, some authors have found a laterally oriented trochlea and others have found a medially oriented trochlea. The groove of implanted prosthetic femoral components always has lateral or neutral orientations. The objectives of the current study were to clarify the controversy found in the literature, to determine whether the trochlear orientation is truly linear, and to determine whether the orientation depends on the size of the femur. The trochleae of 100 human femurs were measured using a three-dimensional measurement system. Detailed analysis of the results indicated that the trochlea is best described as bilinear, with the distal half oriented 0.2 degrees +/- 2.8 degrees laterally and the proximal half oriented 4.2 degrees +/- 3.2 degrees medially. Trochlear orientation was not dependent on bone size. PMID- 12782888 TI - A mechanical comparison and review of transverse, step-cut, and sigmoid osteotomies. AB - Successful incorporation of massive allografts for the treatment of bone deficiency demands maximizing biologic and mechanical factors. These factors have yet to be mastered, as evidenced by the 8% to 17% nonunion and the 5% to 20% fracture rate. The current study addresses the allograft incorporation process by examining the three construct geometries: transverse, step-cut, and sigmoid. Specimens were plated and mounted on a mechanical testing machine. A rotational displacement was applied, and torsional stiffness (N-m/ degrees ), maximum torque (N-m), and maximum displacement ( degrees ) were calculated. The sigmoid osteotomies had a torsional stiffness of 1.90 +/- 0.68 N-m/ degrees and maximum torque of 18.85 +/- 6.63 N-m versus 0.99 +/- N-m/ degrees and 14.48 +/- 2.15 N-m for the transverse osteotomies; and a maximum angular displacement of 11.60 degrees +/- 1.78 degrees versus 5.73 degrees +/- 1.6 degrees for the step-cut osteotomies. The step-cut osteotomies consistently failed at the step-cut corners, which acted as stress risers. Computer-aided solid modeling of the contact surfaces showed that the step and sigmoid osteotomy areas were 74% and 44%, respectively, larger than the transverse osteotomy. The sigmoid osteotomy, created with a template and pneumatic drill, seems to offer a mechanical advantage over the transverse and step-cut osteotomies by increasing stability and contact surface area relative to the transverse osteotomy but reducing the stress-riser effect of the step-cut osteotomy. PMID- 12782889 TI - Intervertebral disc tissue engineering I: characterization of the nucleus pulposus. AB - The characteristics of the nucleus pulposus cells from adult rabbits maintained in in vitro cultures were described in another study. Herein, the authors provide a parallel profile of adult rabbit nucleus pulposus in situ, therefore allowing direct comparisons between in vitro and in situ investigations. Nucleus pulposus specimens from adult rabbits were evaluated using biochemical and immunohistochemical morphologic techniques. The nucleus pulposus from adult rabbits contained cell clusters embedded in proteoglycan-collagen matrix. The cells exhibited a well-defined Golgi system, an extensive endoplasmic reticulum, and a complex vesicular system filled with beaded structures (proteoglycans). Neither necrotic nor apoptotic cells were evident. There was a lack of mitochondria. The extensive extracellular matrix contained amorphous, beaded, and fibrillar components. The fibrillar banding was indicative of Type VI collagen. The nucleus pulposus of adult rabbits expressed aggrecan, collagen Type I and Type II, and CD44, but not collagen Type X and displayed low alkaline phosphatase activity. PMID- 12782890 TI - Intervertebral disc tissue engineering II: cultures of nucleus pulposus cells. AB - The main objective of the current investigation was to regenerate cells of the nucleus pulposus without loss of phenotype. Nucleus pulposus cells were isolated from intervertebral discs from adult rabbits, grown in monolayer culture, and then maintained as a micromass pellet in tube culture. The specimens were evaluated by transmission and light microscopy, reverse transcriptase polymerase chain reaction, and immunohistochemistry. Nucleus pulposus cells proliferated in monolayer culture. When almost confluent, the cells were transferred to a tube and sedimented to form a pellet. The cells reverted to a rounded configuration and formed cell nests surrounded by extensive extracellular matrix, similar to that seen in vivo. These cells did not proliferate. Similar to that observed in situ, cells in pellet culture also expressed aggrecan, CD44, collagen Type II, and collagen Type I, but not collagen Type X, and had low alkaline phosphatase activity. The results of the investigation indicated that nucleus pulposus cells grown in monolayer culture might revert to their original characteristics when transferred to an environment that allows three-dimensional growth, such as upon implantation, a one-step approach. The results also indicated that the two-stage culture procedure might provide an expedient technique to regenerate nucleus pulposus tissue for disc repair. PMID- 12782892 TI - A prescription for the rogue doctor: part I--begin with diagnosis. AB - Among the most serious problems a doctor can have may be those which are the result of a defect of character or a flaw in ethics. Under these circumstances, unacceptable behavior patterns may arise. Examples of unacceptable patterns of behavior include dishonesty, intentionally harming a patient, sexual harassment, and substance abuse. For years, doctors who have these patterns have been handled with kid gloves by those who educate, train, and supervise professionals in the healthcare industry. Counseling, coaching, training, supervising, transfers to less critical disciplines, disciplinary warnings, and offering opportunities to resign have been the typical protocols. Traditionally, outright firing of residents and doctors has been relatively taboo and too radical for the medical profession. Why has this been the case? Reasons may include unwillingness to get involved or to deal with the stress of disciplining a colleague, an unwillingness which often is grounded in fears of retaliation. In a litigious society, fears of slander lawsuits, for example, may be all-too-real. However, the implied paternalism and the practice of protecting doctors' careers by preserving their professional status as practicing doctors have become increasingly problematic. Aside from the fact that it is unethical, allowing problem doctors to continue to practice medicine may have an adverse impact on the well-being of patients and therefore may represent an enormous legal liability for organizations that employ them. In this first of a two-part series, problems that now exist and implementation of a performance management system as a starting point for removing rogues from the system are discussed. A subsequent paper will detail how such a system operates. PMID- 12782891 TI - Microscopic changes at the neuromuscular junction in free muscle transfer. AB - Free muscle transfers do not generate the same force after transfer as that at the original sites. Light and electron microscopy were used to study serially during 30 weeks the changes at the neuromuscular junction after free muscle transfer of the gracilis muscle in the adult Wistar rat. Under light microscopy, after staining with acetylthiocholine the neuromuscular junction showed changes of degeneration with withdrawal of the innervating axon terminal followed by regeneration and reconstitution of the neuromuscular junction. The newly formed neuromuscular junction still lacked the structural detail seen in the control neuromuscular junction, even after 30 weeks. With the electron microscope, mitochondrial swelling and clumping of the synaptic vesicles were followed by withdrawal of the axon terminal from the muscle membrane on denervation. The infolding of the muscle membrane at the neuromuscular junction became less prominent. With reinnervation the ultrastructure of the junction was only partially reestablished with poorly reconstituted primary and secondary folds of the muscle membrane 30 weeks after the transfer. Failure of complete reformation of the ultrastructure of the neuromuscular junction may provide another explanation for failure of full recovery of skeletal muscle function after free muscle transfer. PMID- 12782893 TI - A prescription for the rogue doctor: part II ready, aim, fire. AB - As discussed in Part I of this series, rogue doctors have serious sociopathic behavioral deficiencies which may be the result of a defect of character or a flaw in ethics. Under these circumstances, unacceptable behavior patterns will arise. For years, counseling, coaching, training, supervising, transfers to less critical disciplines, disciplinary warnings, and offering opportunities to resign have been the typical approaches in dealing with rogues; the outright firing of residents and doctors has been relatively taboo. However, preserving a rogue's professional status as a practicing doctor has become increasingly problematic. In Part II of this two-part series, implementation of a performance management system to remove rogues from the system is discussed. This paper will detail exactly how such a system can work. PMID- 12782894 TI - Differentiation of psoas muscle abscess from septic arthritis of the hip in children. PMID- 12782895 TI - Soft tissue mass in a 5-year-old boy. PMID- 12782896 TI - A prepatellar soft tissue mass in a 56-year-old man. PMID- 12782897 TI - Popliteal mass in a 3-year-old boy. PMID- 12782898 TI - Waist circumference, visceral obesity, and cardiovascular risk. PMID- 12782899 TI - Peripheral arterial disease rehabilitation: a review. PMID- 12782900 TI - Stepped care approach to smoking cessation in patients hospitalized for coronary artery disease. AB - PURPOSE: Smoking cessation is an important goal for smokers with coronary artery disease (CAD) because it reduces cardiac morbidity and mortality. Effective interventions for cigarette smokers with CAD exist, but they often are considered to be intensive and expensive. Stepped-care interventions have been proposed as a promising way to allocate smoking cessation treatments in a cost-effective manner. Stepped care refers to the practice of initiating treatment with low intensity intervention and then exposing treatment failures to successively more intense interventions. METHODS: To address the efficacy of this approach, 254 cigarette smokers hospitalized with CAD were provided a brief cessation intervention. The participants then were assigned randomly to either a more intensive stepped-care treatment (counseling and nicotine patch therapy) or no additional treatment. Outcomes were point-prevalent abstinence measured 3 months and 1 year after hospital discharge. RESULTS: Stepped-care treatment increased smoking cessation rates from 42% to 53% during a 3-month follow-up period (P =.05), but showed little effect at the 1-year follow-up assessment, as evidenced by a cessation rate for the minimal intervention group of 36% versus 39% for the stepped-care group (P =.36). CONCLUSIONS: A stepped-care approach to smoking cessation increased short-but not long-term point-prevalent abstinence in patients with CAD. For improvement of long-term effectiveness, refinement of the timing and content of stepped-care interventions needs to occur. PMID- 12782902 TI - Oxygen uptake: when to measure and when to estimate. PMID- 12782901 TI - Predicting peak oxygen consumption during a conservative ramping protocol: implications for the heart failure population. AB - PURPOSE: A significant discrepancy between measured oxygen consumption (VO(2)) (via ventilatory expired gas analysis) and estimated VO(2) (via the imposed workload) frequently is reported in the heart failure (HF) population during symptom-limited exercise testing. The purpose of this investigation was to examine the difference between measured and estimated VO(2) (VO(2) discrepancy) during a highly conservative ramping protocol. METHODS: For this study, 28 subjects with compensated HF (20 men and 8 women; age, 51.1 +/- 14.6 years) and 19 healthy control subjects (age-, gender-, and activity-matched to an HF subgroup) underwent symptom-limited exercise testing (treadmill) with ventilatory expired gas analysis. RESULTS: Peak estimated and measured VO(2) values were significantly higher in the age-, gender-, and activity-matched control group than in the HF group, but the change in measured VO(2) per change in estimated VO(2) (Deltameasured/Deltaestimated VO(2) slope) and the VO(2) discrepancy did not reach statistical significance. Peak estimated VO(2) was a significant predictor of peak measured VO(2) in the overall HF group (R2 = 0.90; P <.001). CONCLUSIONS: Although estimated VO(2) is not considered a replacement for measured VO(2), these results indicate that a highly conservative exercise protocol may allow for a more accurate prediction of peak measured VO(2) via the estimated oxygen cost for a given workload in patients with compensated HF. PMID- 12782903 TI - Training and detraining effects of a combined-strength and aerobic exercise program on blood lipids in patients with coronary artery disease. AB - PURPOSE: The aim of this study was to investigate training and detraining effects on blood lipids and apolipoproteins induced by a specific program that combined strength and aerobic exercise in patients with coronary artery disease (CAD). METHODS: For this study, 14 patients participated in a supervised 8-month training program composed of two strength sessions (60% of 1 repetition maximum) and two aerobic training sessions (60%-85% of maximum heart rate), and 13 patients served as a control group. Blood samples for total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), apolipoproteins A1 (apo-A1) and B (apo-B), and lipoprotein (a) (Lp[a]) were obtained along with muscular strength at the beginning of the study, after 4 and 8 months of training and after 3 months of detraining. RESULTS: The patients in the intervention group showed favorable alterations after 8 months of training (TC, -9.4; TG, -18.6; HDL C, 5.2; apo-A1, 11.2%; P <.05), but these were reversed after 3 months of detraining (TC, +3.7; TG, 16.1; HDL-C, -3.6; apo-A1, -5.5%). In addition, body strength also improved after training (27.8%) but reversed (-12.9%) after detraining (P <.05). The patients in the control group did not experience any significant alterations. CONCLUSIONS: The results indicate that an 8-month training program combining strength and aerobic exercise induces favorable muscular and biochemical adaptations, on TC, TG, HDL-C, and apo-A1 levels, protecting patients with CAD. After 3 months of detraining, however, the favorable adaptations were reversed, underscoring the need of uninterrupted exercise throughout life. PMID- 12782904 TI - Does resistance exercise training reduce the risk of cardiovascular disease? PMID- 12782905 TI - Functional status during immediate recovery after hospitalization for coronary heart disease. PMID- 12782906 TI - Function, eligibility, outcomes, and exercise capacity associated with left ventricular assist devices: exercise rehabilitation and training for patients with ventricular assist devices. PMID- 12782907 TI - Arm exercise capacity and dyspnea ratings in subjects with chronic obstructive pulmonary disease. AB - PURPOSE: This study aimed to compare the metabolic, ventilatory, and dyspnea responses to unsupported arm exercise, supported arm exercise and leg exercise between subjects with chronic obstructive pulmonary disease (COPD) and healthy age-matched controls. METHODS: For this study, 21 subjects with COPD (mean age, 62 +/- 2 years; predicted forced expiratory volume in 1 second [FEV(1)], 37 +/- 3%) and 7 healthy age-matched control subjects (% pred FEV(1) = 109 +/- 5%) were included in the analyses of three incremental exercise tests to peak work capacity: unsupported arm exercise, supported arm exercise (arm ergometry), and leg exercise (cycle ergometry). Work level, oxygen consumption (VO(2)), minute ventilation (V(E)), dyspnea, and rate of perceived exertion were measured each minute. RESULTS: Peak work level and peak VO(2) were significantly reduced in the subjects with COPD for all exercise tests (P <.01 for all), as compared with the control subjects. Within the COPD group, the VO(2) and V(E) at peak exercise were significantly lower for unsupported arm exercise than for both the leg and supported arm exercises (both P <.001). The ratio of V(E) to maximal voluntary ventilation was high for leg exercise (96%), supported arm exercise (91%), and unsupported arm exercise (77%) among the subjects with COPD. At a given percentage of VO(2) peak, dyspnea scores were similar for all the exercise tests. CONCLUSIONS: Ventilatory constraints limit exercise performance in COPD. The lowest amount of work, in terms of VO(2,), was during unsupported arm exercise. Because the subjects with COPD had scores showing similar levels of dyspnea at the same percentage of VO(2) peak, it is suggested that patients be encouraged to reach equivalent dyspnea levels when performing unsupported and supported arm exercise training and leg training. PMID- 12782908 TI - Hit the dyspnea target! PMID- 12782909 TI - Questioning the use of heart rate and dyspnea in the prescription of exercise in subjects with chronic obstructive pulmonary disease. AB - PURPOSE: This study examined the heart rate and dyspnea responses during constant submaximal lower limb endurance exercise in subjects with chronic obstructive pulmonary disease (COPD) to determine the appropriateness of using target heart rate or dyspnea for the prescription of endurance exercise intensity. METHODS: The study participants were 15 men, ages 55 to 75 years, with stable moderate to severe COPD (forced expiratory volume in 1 second, 38.7 +/- 15.6% pred). All the participants completed the incremental shuttle walking test (ISWT) to estimate peak oxygen consumption (VO(2peak)), followed by the endurance shuttle walking test (ESWT) at an intensity equivalent to 60% VO(2peak). Heart rate and dyspnea were monitored before, immediately after, and at 1-minute intervals during each test. RESULTS: The study was completed by 11 subjects. Heart rate and dyspnea increased significantly between 4.5 and 20 minutes during the ESWT (P <.01) despite walking at a constant submaximal workload. Heart rate and dyspnea attained at the end of the ESWT exceeded levels observed during the ISWT at the equivalent workload (P <.05). Four subjects were unable to walk for at least 10 minutes on the ESWT because of severe dyspnea and were withdrawn. CONCLUSIONS: Setting heart rate and dyspnea targets for endurance training at an intensity equivalent to 60% VO(2peak) may be inappropriate for subjects with moderate to severe COPD because heart rate and dyspnea increase independently of workload at this intensity. PMID- 12782910 TI - Fibrosis associated with dopamine agonist therapy in Parkinson's disease. PMID- 12782911 TI - Effectiveness of Gorei-san (TJ-17) for treatment of SSRI-induced nausea and dyspepsia: preliminary observations. AB - Selective serotonin reuptake inhibitors (SSRIs) are apt to cause gastrointestinal adverse events such as nausea and dyspepsia. Gorei-san (TJ-17), which is composed of five herbs (Alismatis rhizoma, Atractylodis lanceae rhizoma, Polyporus, Hoelen, and Cinnamomi cortex), is a Japanese herbal medicine that has been used to treat nausea, dry mouth, edema, headache, and dizziness. The authors investigated the efficacy of TJ-17 for patients who experienced nausea or dyspepsia induced by SSRIs. Twenty outpatients who experienced nausea or dyspepsia induced by SSRIs were recruited for the study. Seventeen patients were female, three were male, and patient age ranged from 21 to 74 years (49.8 +/- 17.0 years). TJ-17 was added to the previous regimen. Nausea and dyspepsia disappeared completely in nine patients, decreased in four patients, decreased slightly in two patients, and did not change in five patients. No adverse events were associated with the addition of TJ-17 in any patient. PMID- 12782912 TI - Excessive daytime sleepiness and levodopa in Parkinson's disease: polygraphic, placebo-controlled monitoring. AB - The authors performed 72-hour polysomnography, including a levodopa-placebo controlled choice reaction time test with continuous monitoring of cardiovascular variables in two patients with Parkinson's disease complaining of excessive daytime sleepiness on levodopa monotherapy. The subacute levodopa challenge was accompanied by physiologic sleep episodes, worsened reaction times, and a reduction in mean blood pressure and heart rate over baseline values, linked temporally to the sleep episodes. PMID- 12782913 TI - Cholinergic-dopaminergic imbalance in Pisa syndrome. AB - Pisa syndrome is a rare type of truncal dystonia. Its development is associated commonly with neuroleptic treatment, but there are rare idiopathic cases or those related to neurodegenerative disorders. Recently, an association between cholinesterase inhibitors and Pisa syndrome has been described. The authors report two patients, one with Alzheimer's disease treated with risperidone and another with Parkinson's disease who presented this kind of dystonia after donepezil initiation. In the first patient the condition resolved after discontinuation of risperidone, and in the second one the condition resolved when donepezil was withdrawn. In patients with pharmacologic or degenerative dopaminergic neurotransmission disorders, cholinergic excess may induce this peculiar type of dystonia. PMID- 12782914 TI - Medications and substances as a cause of headache: a systematic review of the literature. AB - Medication- or substance-induced headache is probably an underrecognized entity with numerous etiologies, including prescribed medication, over-the-counter medication, illicit drugs, anesthetic agents, foods, food additives, beverages, vitamins, inhaled substances, and substances used in diagnostic procedures. The author performs a systemic review of the literature to provide an exhaustive description of the relationship between medications and substances and headaches of various types, along with pathophysiologic mechanisms whenever possible. Suggestions for improved identification of this phenomenon and its avoidance are provided. More scientific evaluation of substances and their possible association with headache is required with almost all substances indicated herein. PMID- 12782916 TI - Risk of serious extrapyramidal symptoms in patients with Parkinson's disease receiving antidepressant drugs: a pharmacoepidemiologic study comparing serotonin reuptake inhibitors and other antidepressant drugs. AB - To compare the risk of occurrence of "serious" extrapyramidal symptoms (EPS) between selective serotonin reuptake inhibitors and other antidepressant drugs in patients with Parkinson's disease (PD), the authors performed a retrospective study using the French Pharmacovigilance Database (i.e., the database recording all serious adverse drug reactions reported in France by physicians to the National French Pharmacovigilance Network). Patients with PD were identified from the case reports including at least one antiparkinsonian drug (except anticholinergics). The authors studied patients with PD exposed to at least one antidepressant (classified as imipraminics, selective serotonin reuptake inhibitors, or "other") drug. EPS were defined as aggravation of the parkinsonian symptoms. Of the76,640 case reports registered in the database between January 1, 1995, and December 31, 2000, 916 were identified as patients treated with at least one antiparkinsonian drug, including 199 treated with antidepressant drugs. Among them the authors found nine case reports of EPS (i.e., 4.5% of the patients with PD treated with at least one antidepressant). The odds ratio for EPS was 2.18 (0.47-11.35) for selective serotonin reuptake inhibitors, 1.17 (0.22-5.50) for imipraminics, and 0.74 (0.10-4.06) for other antidepressants. This study failed to find any significant difference in the occurrence of serious EPS according to the different classes of antidepressant drugs in patients with PD treated with dopaminergic antiparkinsonian drugs. PMID- 12782915 TI - Treatment of neuroleptic-induced akathisia with the 5-HT2A antagonist trazodone. AB - Akathisia is a common and distressful extrapyramidal adverse side effect usually resulting from the use of antipsychotic medications. Early management of akathisia is important because it may be associated with poor treatment response and medication noncompliance. Unfortunately many patients fail to respond to standard management of akathisia. In addition to dopaminergic mechanisms, it has been hypothesized that serotonin may play a prominent role in the pathophysiology of akathisia. Trazodone is an antidepressant agent demonstrating prominent serotonergic antagonistic properties. This open-label pilot study investigates the efficacy of trazodone in the management of akathisia. Nine female patients with a score of at least "mild akathisia" on the Barnes Akathisia Scale, and receiving a stable dose of antipsychotic medication, were administered trazodone, titrated up to a dosage of 100 mg/day over a period of 5 days. The patients demonstrated marked improvement in symptoms of akathisia. In addition, some improvement was noted in symptomatology of anxiety, depression, and psychosis. These observations suggest the use of trazodone as a beneficial and relatively safe medication for the treatment of antipsychotic medication-induced akathisia. Further study in the context of a double-blind, placebo-controlled trial is mandated to substantiate these preliminary findings. PMID- 12782917 TI - High-dose ropinirole in advanced Parkinson's disease with severe dyskinesias. AB - Levodopa (LD) is the gold standard of therapy for Parkinson's disease, but it is commonly associated with motor fluctuations and dyskinesias. Dopamine agonists are often used as adjuncts to LD in an attempt to reduce these complications. In this open-label study the authors investigated the effects of high doses of adjunctive ropinirole in 36 patients with advanced Parkinson's disease and normal cognitive status. The daily dose of ropinirole was increased from 18.4 +/- 3.5 mg to 34.7 +/- 5.5 mg, generally in four separate doses. The daily LD dose was decreased from 734.1 +/- 254.8 mg to 502.8 +/- 228.4 mg. After 12 months 25 patients were still on high doses of ropinirole whereas 11 patients had, after either the emergence of side effects or a worsening of their clinical conditions, decreased or interrupted ropinirole. At 12 months, the daily doses of LD and ropinirole were 489 +/- 243 mg and 34.6 +/- 4.6 mg respectively. There was a significant reduction in the Dyskinesia Rating Scale scores during both ON and OFF periods, indicating a reduction in dyskinesias during ON periods and a reduction in dystonias during OFF periods (p < 0.001). Both the intensity and the hours spent during OFF periods were reduced significantly (p < 0.001). Even though these results need to be confirmed through extended controlled studies, the high-dose dopamine agonist strategy is safe for patients with advanced PD in whom a marked motor response to LD (even at very low doses) is associated with severe dyskinesias, and may be used as a means of delaying surgery or as an alternative to continuous apomorphine infusion. PMID- 12782918 TI - Apomorphine infusion and the long-duration response to levodopa in advanced Parkinson's disease. AB - The authors investigated the long-duration response to levodopa in advanced Parkinson's disease. Eight patients with advanced Parkinson's disease disabled by severe ON/OFF fluctuations treated by chronic daytime subcutaneous apomorphine infusion with supplemental oral levodopa were studied. On day 1, oral levodopa was withdrawn at 4:00 pm and on the following morning subcutaneous apomorphine infusion was continued at the same rate without levodopa therapy. While receiving apomorphine alone, seven of the eight patients turned ON, and their usual dyskinesias returned. The ON phase persisted for 60 to 100 minutes (mean, 185.7 minutes) but then, despite continued, constant-rate apomorphine infusion to stabilize plasma levels, switched to an OFF phase. The authors conclude that the clinical effect of apomorphine is sustained by levodopa long-duration response. This effect is probably the result of postsynaptic mechanisms. In patients with advanced Parkinson's disease, the long-duration response to levodopa is present although slightly diminished. PMID- 12782919 TI - Optimizing levodopa pharmacokinetics: intestinal infusion versus oral sustained release tablets. AB - Continuous duodenal infusion of carbidopa/levodopa has been shown to control motor fluctuations in advanced Parkinson's disease (PD). The authors compared the pharmacokinetics of levodopa and 3-O-methyldopa in patients with advanced PD after administration of an oral sustained-release levodopa preparation and after continuous intestinal levodopa infusion with a new formulation as a gel suspension. A randomized crossover trial was carried out in 12 patients. Carbidopa/levodopa was administered as an oral sustained-release tablet and by nasoduodenal continuous infusion for 3-week periods for each treatment. Plasma levodopa concentrations and motor performance were evaluated every 30 minutes during 3 test days of each treatment period. The average intraindividual coefficient of variation for the plasma levodopa concentrations after oral therapy was 34% and was significantly lower (14%, p < 0.01) during continuous infusion. Hourly video evaluations showed a significant increase in ON time during infusion and a significant decrease in OFF time and dyskinesia. Continuous intraduodenal delivery of a new carbidopa/levodopa formulation offers a means for markedly improved control of motor fluctuations in late stages of PD. PMID- 12782920 TI - First clinical evaluation of ganstigmine in patients with probable Alzheimer's disease. AB - The objective of this study was to evaluate the safety, tolerability, maximum tolerated dose, pharmacokinetics, and pharmacodynamics of five fixed doses of ganstigmine (CHF 2819) in patients with probable Alzheimer's disease (AD). This randomized, double-blind, placebo-controlled trial evaluated five dose levels (5, 7.5, 10, 12.5, and 15 mg) administered orally once daily for 7 days. Adverse events and continuous telemetry were collected on successive panels of six patients (five active, one placebo). Acetylcholinesterase, butyrylcholinesterase, and plasma drug levels were measured. A total of 29 patients were randomized and 18 completed the study. A total of seven patients, including five of five in the 12.5-mg panel, discontinued because of adverse events. Four patients were withdrawn administratively from the first panel while an episode of atrial fibrillation (the only serious adverse event) was investigated. This panel was then repeated. Mild, transient headache or nausea were the most commonly reported adverse events. Multiple moderate adverse events in the 12.5-mg panel (including nausea, vomiting, and anorexia) led to the decision not to proceed with a 15-mg panel. Ten milligrams was determined to be the maximum tolerated dose. Ganstigmine exhibited nonlinear pharmacokinetics, was absorbed rapidly, and reached peak concentrations within 1 hour. Acetylcholinesterase inhibition was dose dependent and lasted as long as 24 hours. Ganstigmine, a novel cholinesterase inhibitor, was well tolerated within a dosing range of 5 to 10 mg. Once-daily dosing is supported by data on acetylcholinesterase inhibition. PMID- 12782921 TI - What is Vasculitis? PMID- 12782922 TI - Radiosensitive orbital inflammation associated with temporal arteritis. AB - A 75-year-old woman developed acute loss of vision in the OD, ipsilateral periocular pain, an afferent pupillary defect, sectoral optic disc edema, and later ipsilateral proptosis and an intraconal mass. She denied any symptoms of temporal arteritis, and a sedimentation rate was normal. Orbital biopsy demonstrated chronic granulomatous inflammation with perivasculitis. A temporal artery biopsy disclosed findings consistent with temporal arteritis. Following 2000 cGy of external beam radiation, her visual function and orbitopathy completely resolved. This unusual presentation of orbital inflammation in association with temporal arteritis demonstrates that pathologic findings of temporal arteritis may be clinically nonspecific and that external beam radiation may be an effective therapy in this setting. PMID- 12782923 TI - Late ipsilateral recurrence of ischemic optic neuropathy in giant cell arteritis. AB - A patient with arteriosclerosis, diabetes mellitus, and giant cell arteritis (GCA) treated continuously with low-dose prednisone developed anterior ischemic optic neuropathy (AION) at 5 and 13 months after clinical diagnosis of GCA. At the time of late recurrent AION, there were no systemic symptoms or elevations in acute phase reactants to signal active arteritis, yet temporal artery biopsy disclosed dramatic inflammation, forcing the presumption that the infarct was arteritic. Recurrent systemic symptoms and elevation of acute phase reactants are not reliable warning signs of reactivated GCA. In patients at high risk for corticosteroid complications, late biopsy may be a reasonable guide to corticosteroid weaning. PMID- 12782924 TI - Linear magnetic resonance enhancement and optic neuropathy in primary angiitis of the central nervous system. AB - A 38-year-old woman developed incoherent mentation, tremor, ataxia, and bilateral optic disc edema with mildly depressed visual acuity, nerve fiber bundle defects, and a left afferent pupillary defect. Magnetic resonance imaging of the brain disclosed striking linear contrast enhancement radiating from the ventricular borders. Lumbar puncture showed a normal opening pressure with a lymphocytic pleocytosis and elevated protein. On the basis of these findings, the initial diagnosis was viral or post-viral meningoencephalitis and the patient was not treated. During the next 4 weeks, her condition worsened. A brain and meningeal biopsy disclosed findings typical of primary angiitis of the central nervous system. With aggressive treatment, her neurologic status and magnetic resonance imaging normalized and her optic neuropathy improved markedly. Optic neuropathy and linear magnetic resonance imaging enhancement should be recognized as features of primary angiitis of the central nervous system. PMID- 12782925 TI - Persistent severe visual and electroretinographic abnormalities after intravenous Cisplatin therapy. AB - A 55-year-old man inadvertently received four times the intended dose of intravenous cisplatin as part of a chemotherapeutic salvage regimen for non Hodgkin lymphoma. Immediately after treatment, he developed bilateral irreversible visual loss. Visual acuity was 20/300 in OU and visual fields showed central scotomas bilaterally. Although the fundus examination findings were normal, an electroretinogram showed markedly reduced a-wave amplitudes and absent b-waves. At autopsy 8 months later, photoreceptors appeared normal. Splitting of the outer plexiform layer was present, consistent with loss of the ERG b-wave. This is the first reported case of persistent visual loss from intravenous cisplatin toxicity and the first case to describe ocular histopathologic findings. PMID- 12782926 TI - Surgical management of skew deviation. AB - There are no published data on the outcomes of realignment surgery for skew deviation. A retrospective chart review disclosed 10 patients who had undergone surgical correction of skew deviation by three surgeons at a single institution between 1991 and 2002. Nine of 10 patients had satisfactory relief of diplopia with an acceptable field of single binocular vision. Vertical rectus recession or resection was the most common procedure. Four patients required more than one procedure. For nonalternating hypertropias, resection of the inferior rectus muscle or recession of the superior rectus muscle of the hypertropic eye was successful. For alternating hypertropia, resection of both inferior rectus muscles was successful. Oblique muscle surgery was not associated with good outcomes. PMID- 12782927 TI - Progressive visual loss because of a suprasellar pneumatocele after trans sphenoidal resection of a pituitary adenoma. AB - A 63-year-old man who underwent uneventful trans-sphenoidal resection of a pituitary adenoma with fat packing complained postoperatively of progressive binocular visual acuity loss. Neuroimaging showed a suprasellar pneumatocele compressing the optic chiasm and a communication between the sphenoid sinus and the sella. After a second trans-sphenoidal procedure to remove the air and fully pack the sphenoid sinus, visual acuity recovered dramatically. A rare complication of trans-sphenoidal surgery for pituitary adenoma, suprasellar pneumatocele probably forms through a ball-valve mechanism that results from incomplete packing of the sellar floor. This case highlights the need for effective sphenoid sinus packing and for ophthalmic monitoring after trans sphenoidal surgery. PMID- 12782928 TI - Environmental tilt illusion as the only symptom of a thalamic astrocytoma. AB - A 14-year-old girl experienced two episodes of environmental tilt illusion. During both episodes, which lasted less than 1 minute, she perceived all objects within view as rotated 45 degrees clockwise. There were no auras, accompanying symptoms, or sequelae. Neuro-ophthalmic examination findings were normal except for a right relative afferent pupil defect (RAPD). Imaging disclosed a cystic mass in the left posterior thalamus with compression of the brachium of the left superior colliculus. Stereotactic biopsy revealed a pilocytic astrocytoma. This is the first case documenting environmental tilt illusion as an isolated symptom of a thalamic lesion. Disruption of vestibular connections between the posterior thalamus and the posterior parietal cortex may be the cause of this visual perceptive disorder. PMID- 12782929 TI - Leptomeningeal enhancement and venous abnormalities in granulomatous angiitis of the central nervous system. AB - A 68-year-old woman with a relatively acute onset of right homonymous hemianopia, Gerstmann syndrome, and global cognitive failure was found to have a lymphocytic pleocytosis and elevated protein on spinal fluid examination and displayed marked meningeal enhancement on magnetic resonance imaging and dilated cortical venules on cerebral angiography. Brain and meningeal biopsy disclosed a necrotizing granulomatous inflammation of small and medium-sized subarachnoid vessels. The brain parenchyma was normal. The angiographic presence of venous abnormalities, the lack of observable angiographic arterial involvement, and the lack of parenchymal pathology are distinctly unusual in granulomatous angiitis of the central nervous system. This case, therefore, extends the pathologic and imaging spectrum of this disorder. PMID- 12782930 TI - Giant cavernous malformation of the occipital lobe. AB - A 15-year-old boy who developed severe headaches and an incomplete homonymous hemianopia was found to have a large, well-circumscribed, multilobulated intracranial mass in the contralateral occipital lobe. The initial impression was that of a low-grade glioma or a vascular malformation. When the lesion increased in size and complexity, concern arose about the possibility of a malignant glioma. Upon craniotomy, it proved to be a giant cerebral cavernous malformation. This case is remarkable in that most cavernous malformations do not become symptomatic before the third decade of life and rarely attain such a large size. PMID- 12782931 TI - Self-inflicted blindness and Brown-Sequard syndrome. AB - A 30-year-old paranoid schizophrenic man suffered a psychotic episode while flying on an airplane, locked himself in the bathroom, detached the temples of his sunglasses, and stabbed them deeply into both medial orbits. He then secured one temple into the door hinge and rammed the back of his neck repeatedly against it. The injuries caused no light perception from optic nerve trauma and a Brown Sequard hemitransection of the spinal cord. PMID- 12782932 TI - Pathogenesis of nonarteritic anterior ischemic optic neuropathy. AB - Based on histopathology, electron microscopic corrosion cast studies, optic nerve blood flow studies, and clinical data, the pathogenesis of idiopathic nonarteritic ischemic optic neuropathy includes the following features: (1) structurally crowded optic discs are predisposed; (2) laminar and retrolaminar regions are the most common locations for infarction; (3) there is flow impairment in the prelaminar optic disc during the acute phase; (4) lack of consistent choroidal flow impairment and the retrolaminar location of infarcts suggest vasculopathy within or distal to the paraoptic branches of the posterior choroidal arteries; (5) diabetes is the most consistently identified vasculopathic risk factor; (6) impaired autoregulation of the disc circulation by atherosclerosis, with a possible contribution from serotonin and endothelin mediated vasospasm, may play a role; and (7) progression may be caused by secondary cell death after the initial ischemic insult or compression from cavernous degeneration and mechanical axonal distortion. PMID- 12782934 TI - Bilateral distribution of the end branches of the pontine paramedian branches of the basilar artery. PMID- 12782933 TI - Video electroencephalogram monitoring during paroxysmal upside down reversal of vision. PMID- 12782935 TI - Asymmetric myopia developing after optic neuritis. PMID- 12782936 TI - Migraine-like visual hallucinations in occipital lesions of cysticercosis. PMID- 12782937 TI - Thalidomide analogs as emerging anti-cancer drugs. AB - Recently, it has been demonstrated that a number of novel thalidomide analogs possess anti-cancer properties due to their T cell co-stimulatory, anti angiogenic and/or anti-inflammatory effects. Based on such effects, a class of thalidomide analogs known as Immunomodulatory Drugs (IMiDs) have recently entered into phase I clinical trials for the treatment of a number of cancers. The lead IMiD CC-5013 (referred to clinically as REVIMID) is now entering phase III clinical trials for multiple myeloma and metastatic melanoma, while CC-4047 (ACTIMID) is currently under investigation in phase I/II and II trials for multiple myeloma and prostate cancer, respectively. The other group of compounds, classified as Selective Cytokine Inhibitory Drugs (SelCIDs), do not co-stimulate T cells, but have anti-inflammatory and anti-angiogenic properties. Moreover, a subset of SelCIDs has been found to possess direct anti-tumor activity both in vitro and in vivo. This minireview highlights the various mechanisms of action associated with these compounds and their subsequent clinical development. The enhanced efficacy and lower side-effect profiles of the analogs in comparison to thalidomide make the use of these agents very attractive as novel anti-cancer agents. PMID- 12782938 TI - Treosulfan and gemcitabine in metastatic uveal melanoma patients: results of a multicenter feasibility study. AB - No effective treatment currently exists for metastatic uveal melanoma. However, recent results obtained by an ATP-based tumor chemosensitivity assay have shown consistent activity of treosulfan+gemcitabine in up to 80% of tumor specimens tested. In this study we describe the first clinical results observed with this drug combination at different European centers in patients with metastatic uveal melanoma. Clinical case series of patients with metastatic uveal melanoma were treated with treosulfan+gemcitabine at seven different centers. Fourteen patients, 13 previously untreated and one pretreated with chemoimmunotherapy, were included in the study. Patients received treosulfan+gemcitabine in four different dose regimens. The response rates, progression-free and overall survival, and toxicity were evaluated. The analysis of 14 patients revealed one complete response, three partial responses and a stable disease in eight cases. The objective response rate was 28.6%, the median overall survival was 61 weeks [95% confidence interval (CI) 54-133 weeks], the progression-free survival was 28.5 weeks (95% CI 13-62 weeks) and the 1-year survival rate was 80%. The drugs were well tolerated. The most common side-effects were leuko- and thrombocytopenia. These preliminary results suggest potential therapeutic benefit of treosulfan+gemcitabine treatment in metastatic uveal melanoma and warrant further controlled studies. PMID- 12782939 TI - Hematological side-effect profiles of individualized chemotherapy regimen for recurrent ovarian cancer. AB - The long-term results for patients with recurrent ovarian cancer (ROC) are poor. There is a need to optimize treatment strategies to improve outcome by avoiding ineffective regimens which are often associated with exacerbated side-effects. Individualized chemotherapy regimens guided by a chemosensitivity assay (ATP tumor chemosensitivity assay) have already been used successfully to direct chemotherapy. Taking the results of this assay into account, application of drug combinations appears more advisable. Here we present a systematic evaluation of toxicities seen with individualized chemotherapy for ROC. A total of 62 patients who received 314 cycles of antineoplastic therapies were evaluated. Three single agents (topotecan, paclitaxel and gemcitabine) and five combinations (cisplatin/gemcitabine, carbopatin/gemcitabine, gemcitabine/treosulfan, mitoxantrone/paclitaxel and carboplatin/paclitaxel) were examined. With respect to myelotoxicity, most single agents except topotecan revealed favorable results in comparison to drug combinations. However, this observation lacks statistical significance. Generally, severe myelosuppression was rare. The highest incidence of leukopenia was seen in regimens with mitoxantrone/paclitaxel or gemcitabine/treosulfan, respectively. Thrombocytopenia accompanied most commonly a topotecan therapy. In the present study combination regimens tend to be more toxic than monotherapies. When response rates are comparable, empirically chosen treatment combination therapies should only be practiced in carefully planned clinical studies. PMID- 12782940 TI - Long-term results of paclitaxel in FIGO stage III ovarian carcinoma. AB - The combination of platinum and paclitaxel is the standard treatment of advanced ovarian carcinoma; however, recent studies have questioned the actual role of the combination as compared to either of the two agents alone. We report an open label, two-center, phase II study of upfront paclitaxel for patients with histological diagnosis of stage III ovarian carcinoma. Treatment consisted of paclitaxel at 175 mg/m2 administered in a 3-h infusion every 21 days. Response was evaluated after the third course by either laparoscopy or exploratory laparotomy. Patients with stable or progressive disease discontinued treatment, whereas responding patients continued treatment until a maximum of six courses. Response, toxicity, time to progression (TTP) and survival were evaluated. From November 1993 to December 1995, 30 patients were accrued. All patients underwent primary cytoreduction; 17 (57%) and 13 (43%) patients had residual tumors <2 and >2 cm, respectively. Of 27 patients evaluable, objective responses were seen in 18 (66.4%) (95% CI 49.5-83.2)--12 complete (45%) and six partial (22%). Four patients had stable disease (15%) and five (18%) patients progressed. A total of 149 courses were administered to 30 patients, median 4 (range 1-6). Grade 3/4 neutropenia was seen in 13% of courses, peripheral neuropathy, myalgia and arthralgia were frequent, but transitory and relieved with analgesics. At a median follow-up time of 44.5 months (0-99) the TTP and median survival were 16.6 and 43.1 months, respectively. We conclude that single-agent paclitaxel is an effective and well-tolerated first-line treatment for advanced ovarian carcinoma. PMID- 12782941 TI - Phase I population pharmacokinetics of irofulven. AB - Our aim was to develop a population pharmacokinetic model for irofulven and to assess covariates that might affect irofulven pharmacokinetics. Irofulven was administered by 5- or 30-min i.v. infusion to cancer patients during a phase I study. Blood samples were collected over 4 h. Plasma samples were analyzed to quantitate irofulven by high-performance liquid chromatography. Population pharmacokinetic analysis was performed using a non-linear mixed effects modeling program, MP2. Fifty-nine patients were available for pharmacokinetic analysis. Irofulven plasma concentration-time profiles were best described by a two compartment pharmacokinetic model. Clearance and central volume of distribution were not significantly influenced by individual characteristics, i.e. body weight (BW), body surface area (BSA), age and gender. Final parameter estimates of clearance and central volume of distribution were 616 l/h and 37 l, respectively, resulting in a very short terminal half-life of less than 10 min. A relatively high level of variability was observed in irofulven pharmacokinetics, which was mainly due to a significant residual variability, 39%. For a 30-min irofulven infusion, the optimal sampling schedule for clearance estimation using the Bayesian method was the three time points 0.35-0.45, 0.80 and 1-1.2 h from the beginning of a 30-min infusion. We conclude that after i.v. infusion of irofulven, plasma clearance was high and not dependent upon patient age, gender, BSA or BW. PMID- 12782943 TI - Distribution of paclitaxel in plasma and cerebrospinal fluid. AB - Our objective was to assess the distribution of paclitaxel in plasma and cerebrospinal fluid (CSF) in a cancer patient, and evaluate the role of the formulation vehicle Cremophor EL (CrEL) in drug distribution. Analysis of paclitaxel concentrations in CSF was performed using a triple-quadrupole mass spectrometric assay with electrospray ionization. Total and unbound paclitaxel levels in plasma were measured by liquid chromatography and equilibrium dialysis, respectively, and CrEL concentrations were determined by a colorimetric dye binding microassay. Clinical samples were obtained from a 54-year-old female with breast cancer receiving a weekly regimen of paclitaxel (dose 60 mg/m2). The disposition of total paclitaxel in plasma was characterized by a bi-exponential elimination (terminal half-life 9.17 h) and a total clearance of 19.4 l/h/m2. The fraction of unbound paclitaxel in plasma ranged from 7.6 to 12.4% (unbound drug CL 176 l/h/m2). The plasma clearance of CrEL was 0.332 l/h/m2, whereas CrEL levels were undetectable in CSF (below 0.5 microl/ml). Concentrations of paclitaxel in CSF (range 45.5-162 pg/ml) and unbound CSF:unbound plasma concentration ratios (range 0.093-9.53%) progressively increased up to 24 h, with a mean unbound drug fraction in CSF of 84+/-3.6% (range 81-88%). These findings indicate that there is substantial distribution of paclitaxel to CSF. Since the fraction of unbound paclitaxel is different between plasma and CSF, measurement of unbound paclitaxel is required to accurately assess the extent of drug penetration. PMID- 12782942 TI - Escalating doses of paclitaxel and epirubicin in combination with cisplatin in advanced ovarian epithelial carcinoma: a phase I-II study. AB - Our objective was to identify a new active three-drug combination regimen consisting of paclitaxel (PTX), epirubicin (EPI) and cisplatin as first-line line chemotherapy for advanced ovarian carcinoma. A phase I study was carried out to evaluate the dose-limiting toxicity (DLT) and the maximally tolerated dose (MTD) of PXT and EPI in combination with a fixed dose of cisplatin every 4 weeks. Side effects were recorded according to the NCI Common Toxicity Criteria. Patients were treated in cohorts of three with fixed-dose cisplatin 80 mg/m2 and EPI 80- >100 mg/m2 and PXT 100-->160 mg/m2 until DLT was reached. Once MTD was identified, a single-step phase II study was therefore carried out to test the clinical activity and panel of toxicity of such regimen. Objective responses were recorded according to the WHO criteria. Time to progression and overall survival (OS) were secondary endpoints. The DLT was myelosuppression and, in more detail, febrile neutropenia, which occurred at the fifth dose level (PTX 140 mg/m2, EPI 100 mg/m2 and cisplatin 80 mg/m2) in two out of three patients. Other side effects were grade 3 mucositis in two out of three patients and grade 3 anemia in one case. The combination of cisplatin 80 mg/m2 plus EPI 80 mg/m2 and PCT 140 mg/m2 every 4 weeks was considered as the MTD. In the phase II study a complete response was observed in six patients (33%) and a partial response in nine cases (50%) for an overall response rate of 83% [95% confidence limits (CL) 59-96%]. Median time to progression of patients with measurable disease was 16.4 months. Median OS was not reached after a follow-up of 42 months. This study demonstrated that PTX and EPI can be safely administered in combination with cisplatin to fit patients with advanced epithelial ovarian carcinoma. The three-drug regimen of cisplatin 80 mg/m2, EPI 80 mg/m2 and PTX 140 mg/m2 every 4 weeks is very active, at least in terms of objective response rate. This level of activity overlaps with the 95% CL of the activity of cisplatin alone; however, it does encourage future trials of the combination. PMID- 12782944 TI - Heterogeneity of chemosensitivity of colorectal adenocarcinoma determined by a modified ex vivo ATP-tumor chemosensitivity assay (ATP-TCA). AB - Advanced colorectal cancer (CRC) has a poor prognosis with a 5-year survival of only 5% despite treatment with chemotherapeutic agents. Response rate and overall survival varies little between the commonly used single agents, although combinations achieve better outcomes. It is well established that considerable heterogeneity exists between cancers of the same tissue type, but it has been difficult to establish this for CRC. We therefore investigated the heterogeneity of chemosensitivity in CRC using a modified version of the ex vivo ATP-tumor chemosensitivity assay (ATP-TCA) capable of handling infected tumor tissue. Fifty three specimens of primary solid or malignant effusions of CRC were tested, of which 46 (87%) were evaluable. There were considerable differences in sensitivities between individuals. The most active single cytotoxic agents in the assay were identified as 5-fluorouracil, irinotecan and mitomycin C (MMC). Cells were exposed to combinations of drugs added simultaneously at the same concentrations tested as single agents. All drug combinations achieved greater growth inhibition than drugs used alone. MMC+gemcitabine was found to be the most effective combination in 83% of specimens. The ATP-TCA has previously been shown to be a good predictor of response to chemotherapy in other tissue types. The degree of heterogeneity demonstrated from these results suggests that the ATP-TCA could be used to identify patients who might benefit from specific chemotherapeutic agents alone or in combination. PMID- 12782945 TI - Phosphodiesterase 4 in osteoblastic osteosarcoma cells as a potential target for growth inhibition. AB - Reverse transcription-polymerase chain reaction (RT-PCR) analysis revealed that HOSM-1 cells, an osteosarcoma cell line established from human mandible, expressed mRNA for osteoblastic markers, such as alkaline phosphatase, osteonectin, osteocalcin and parathyroid hormone receptor, thus exhibiting an osteoblastic phenotype. We have investigated a possible role of cyclic nucleotide phosphodiesterases (PDEs) in osteosarcoma cells. RT-PCR analysis revealed that HOSM-1 cells expressed mRNA for PDE4A, 4B and 4C. In addition, rolipram, a specific inhibitor of PDE4, inhibited HOSM-1 cell proliferation. The finding that PDE4 is involved in proliferation of osteosarcoma cells suggests the possibility that PDE4 may be a new target for antitumor therapy. PMID- 12782946 TI - The effects of doxorubicin on apoptosis and adhesion molecules of normal peripheral blood leukocytes-an ex vivo study. AB - This ex vivo study was designed to evaluate the effect of doxorubicin (Dox) on normal peripheral blood leukocytes (PBL) in terms of apoptosis and membranal expression levels of adhesion molecules. Blood was drawn immediately prior to and after Dox administration from 21 breast cancer patients, and incubated at room temperature for 24 h. Flow cytometry was employed in analysis of apoptosis with Annexin-V and protein membranal expression levels with monoclonal antibodies to CD49d, CD18, CD11a-c and CD63. Dox induced statistically significant apoptosis in all three major PBL subpopulations (p<0.01). Between 70 and 90% of samples underwent apoptosis in all PBL subgroups. No significant change was observed in the membranal level of CD63, CD49d and CD11a-c after chemotherapy in any PBL subpopulation. However, a significant reduction in the membranal level of CD18 was demonstrated in polymorphonuclear cells after Dox (p<0.005) both in apoptotic and non-apoptotic cells (p<0.05), suggesting a direct effect of Dox rather than an apoptosis-associated phenomenon. We observed the expected leukopenia 10 days after Dox administration with no correlation to apoptosis, suggesting that leukopenia by Dox is largely attributed to toxicity of blood progenitors. PMID- 12782947 TI - STD transmission dynamics: some current complexities. 2002 Thomas Parran Award Lecture. PMID- 12782948 TI - Evaluation of a new rapid diagnostic kit (FemExam) for bacterial vaginosis in patients with vaginal discharge syndrome in The Gambia. AB - BACKGROUND: Diagnosis of bacterial vaginosis (BV) in resource-poor primary health care settings is often overlooked; there is a need for a cheap, rapid, objective point-of-care diagnostic test. GOAL: The goal was to determine the prevalence of BV and to evaluate the performance of a new commercial diagnostic test kit in a developing country environment. STUDY DESIGN: Vaginal and cervical swabs were collected from 230 consecutive women attending a genitourinary medicine clinic with reported symptoms of vaginal discharge and/or itching. Etiological testing was carried out. BV was diagnosed on the basis of the Nugent score, the Amsel clinical criteria, and results of FemExam card tests. Card 1 is for pH and amines, and card 2 measures proline iminopeptidase (PIP) activity. RESULTS: BV prevalence was 47.9% according to the Nugent score. When compared with the Nugent score, the Amsel clinical criteria had a sensitivity of 77.9% and specificity of 58.4%, FemExam card 1 had a sensitivity of 71.4% and specificity of 72.8%, FemExam card 2 had a sensitivity of 70% and specificity of 81.0%, and FemExam cards 1 and 2 combined had a sensitivity of 91.0% and specificity of 61.5%. Cost per patient and cost per true case detected ranged from US $0.74 and US $1.54, respectively, for Gram stain diagnosis, to US $8.32 and US $18.49 for the FemExam two-card method. CONCLUSIONS: In a setting where BV was frequently associated with vaginal discharge, the FemExam test compared favorably with conventional clinical diagnosis, and it has the advantage of being rapid, less subjective, and easily performed. Cutting its cost would provide wider accessibility in developing countries. PMID- 12782950 TI - Outbreak of Neisseria gonorrhoeae in Northern Alberta, Canada. AB - BACKGROUND: In January 2001 we investigated an outbreak of Neisseria gonorrhoeae in a northern region of Alberta, Canada, and here we report on the epidemiology of the outbreak. GOAL: The goal was to examine the outbreak etiology and make recommendations for strengthening regional STD programs. STUDY DESIGN: Provincial STD notification forms were reviewed to identify cases, and a case-control study was undertaken to identify risk factors for infection. RESULTS: Gonorrhea was reported among 81 individuals, aged 15 to 60 years, between January 1999 and March 2001 in 8 neighboring communities. Attendance at a public bar in one community was associated with infection (P < 0.01). Cases were predominantly Aboriginal (96.3%) and aged less than 30 years (77.6%), and at least 39% of cases reported multiple sex partners. Casual partnering and extended sexual networks are believed to have influenced the spread of infection. CONCLUSION: This study emphasizes the importance of site-specific health interventions to effectively target at-risk individuals at high-risk locations, with preventive measures aimed at members of high-risk sexual networks. Effective interventions must ensure the availability of and access to appropriate health services for all residents of northern regions in Alberta. PMID- 12782949 TI - Partner notification for HIV and STD in the United States: low coverage for gonorrhea, chlamydial infection, and HIV. AB - BACKGROUND: Little is known about the scope of current public health partner notification (PN) activities in the United States. GOAL: The goal of the study was to define what PN services U.S. health departments provide in areas with high STD/HIV-related morbidity. STUDY DESIGN: The study involved a survey of STD program staff members in U.S. areas with the highest reported rates of infectious syphilis, gonorrhea, chlamydia, and HIV in 1998. RESULTS: Staff members of 60 (77%) of 78 health departments provided data. PN interviews were conducted with 7583 (89%) of 8492 cases of syphilis, 23,097 (17%) of 139,287 cases of gonorrhea, and 26,487 (12%) of 228,210 cases of chlamydia. In areas with mandatory HIV reporting, 4375 (52%) of 8328 persons infected with HIV were interviewed for PN. CONCLUSIONS: Except for patients with syphilis, public health PN services affect only a minority of persons with STD or HIV infection in high-morbidity areas of the United States. PMID- 12782951 TI - The cost-effectiveness of single-dose azithromycin for treatment of incubating syphilis. AB - BACKGROUND: Treatment of incubating syphilis with intramuscular benzathine penicillin in exposed sex partners is not always practical in the field, and exposed partners may not adhere to referrals for treatment at clinical facilities. The availability of a single-dose oral therapy could increase the number of partners treated and reduce future infections. GOAL: The goal of the study was to evaluate the cost-effectiveness of directly observed oral administration of azithromycin as an alternative to referral for treatment with benzathine penicillin. STUDY DESIGN: Using published probability and cost estimates, we constructed a decision-analysis model to compare the direct costs and effectiveness of field treatment with azithromycin (1-g single dose) versus referral for standard benzathine penicillin therapy. RESULTS: At public-sector pricing ($11.50 U.S. dollars), directly observed field treatment with azithromycin is cost-saving from both the program and healthcare system perspectives at efficacy levels as low as 75%. Azithromycin therapy is cost saving at the wholesale price of $17.32 U.S. dollars (sachet formulation) when efficacy is at least 90%. The more expensive tablet formulation (average wholesale price of $27.89 U.S. dollars) is not cost-saving from a program perspective, but it remains cost-saving from a healthcare system perspective if efficacy rates are at least 90%. Azithromycin therapy (1-g single dose) will result in fewer cases of early syphilis among exposed partners, provided that the drug's efficacy is at least 87%. CONCLUSIONS: Azithromycin is a cost-effective alternative treatment for incubating syphilis in settings where standard intramuscular therapy is not practical. PMID- 12782952 TI - Patient-delivered therapy for chlamydia: putting research into practice. AB - BACKGROUND: In 2001 California State lawmakers authorized patient-delivered therapy for sex partners of patients with chlamydial infection. Several studies have suggested that patient-delivered therapy is effective in the treatment of infected partners. GOAL: The goal was to inform clinicians of the effectiveness and feasibility of patient-delivered therapy for the management of partners of sexually transmitted disease cases. STUDY DESIGN: A review of studies regarding patient-delivered therapy was performed. A descriptive cross-sectional analysis of clinic data was also done. The proportion of patients with chlamydia receiving therapy for partners and stratification by year and selected characteristics were evaluated with statistical analysis software (SAS). RESULTS: Studies suggested patient-delivered therapy decreased the incidence of chlamydia infection and the risk of reinfection from an untreated partner. The annual proportion of cases in which patient-delivered therapy occurred at the San Francisco STD Clinic was approximately 23%. CONCLUSIONS: Patient-delivered therapy is a beneficial and feasible addition to partner notification in the management of chlamydia. Expansion of patient-delivered therapy should be considered seriously in public health policy and clinical care. PMID- 12782953 TI - Comparison of STD burden and risk among men with and without regular doctors attending a southern urban STD clinic. AB - BACKGROUND: Studies addressing health-seeking behaviors among men at risk for STD are few; this study examines how having a regular doctor might influence risk taking or STD prevalence. GOAL: The goal was to examine demographic, sexual, and health-seeking characteristics of male STD clinic attendees. STUDY DESIGN: Randomly selected men attending an STD clinic answered a questionnaire regarding the aforementioned characteristics. The men were stratified according to whether they reported having a regular doctor. RESULTS: Of 467 men, 32% reported a regular doctor. These men were more likely to be older, to be better educated, and to state they would seek care for a medical problem at a doctor's office. The two groups did not differ in STD history or prostitute exposure; men without doctors were more likely to have urethritis and STD, although the men with doctors had substantial STD rates. CONCLUSION: Public health clinics remain an important safety net for the management of STD, even for men who report a regular doctor. PMID- 12782954 TI - Addition of treatment for trichomoniasis to syndromic management of urethritis in Malawi: a randomized clinical trial. AB - BACKGROUND: Male urethritis is generally treated syndromically, but failure of empirical treatment is common. GOAL: The study goal was to evaluate the addition of metronidazole to the syndromic management of urethritis in Malawi in a randomized clinical trial. STUDY DESIGN: Men with urethritis were randomized to receive either 2 g of metronidazole by mouth or placebo, in addition to standard care for urethritis (i.e., a single intramuscular dose of 240 mg gentamicin and 100 mg doxycycline twice daily for 7 days). The primary endpoints of the study included measurement of the effects of treatment on Trichomonas vaginalis, signs and symptoms of urethritis, and the concentration of HIV RNA in semen in dually infected subjects. RESULTS: The overall prevalence of T vaginalis was 17.3% (71/411), and treatment with metronidazole cleared 95% of culture-positive infections, compared with 54% clearance among men receiving placebo (P = 0.006). Prevalence of persistent urethritis was observed in approximately 16% of both groups at the end of 1 week (29/179 of those receiving metronidazole versus 29/187 in the placebo group; P = 0.86). For a subset of HIV-infected men with trichomoniasis, the seminal plasma HIV RNA concentration was higher than in a group of HIV-positive control subjects (median copies/mL:35,000 vs. 1800 P = 0.06) [correction]. CONCLUSION: In areas with a high prevalence of trichomoniasis, the addition of metronidazole to the syndromic management of male urethritis can eliminate infection with T vaginalis and may help to reduce the transmission of HIV. Such treatment should be strongly considered as part of empirical therapy for urethritis in men in Malawi and places where T vaginalis infection in men is common. PMID- 12782955 TI - Alarming increase in ciprofloxacin- and penicillin-resistant Neisseria gonorrhoeae isolates in New Delhi, India. AB - BACKGROUND: Resistance of ciprofloxacin has been reported in several regions of the world, including India. In India, ciprofloxacin is still being used as single dose treatment for gonorrhea. GOAL: The aim of the study was to determine the antimicrobial susceptibility of Neisseria gonorrhoeae isolates from patients with acute gonococcal urethritis in New Delhi. STUDY DESIGN: By means of disc diffusion, we determined the susceptibility profiles of N gonorrhoeae isolates, determined the MICs of ciprofloxacin, penicillin, and ceftriaxone, and compared our data with previous findings. RESULTS: On the basis of MIC values, 35.3% and 52.9% of strains were found to be resistant and less sensitive, respectively, to penicillin; 67.3% and 28.2% strains were observed to be resistant and less sensitive, respectively, to ciprofloxacin. Only one isolate (5.9%) was found to be less sensitive to ceftriaxone. CONCLUSION: The significant increase in ciprofloxacin resistance in the current study indicates that resistance has developed under selective antibiotic pressure. PMID- 12782956 TI - Response to "Increased reporting of male-to-male sexual activity in a national survey". PMID- 12782957 TI - Fluoroquinolones, gonorrhea, and the CDC STD treatment guidelines. PMID- 12782959 TI - Using qualitative methods to design an epidemiologic study on sexually transmitted diseases in female prisoners. PMID- 12782960 TI - NOTCH4 gene promoter polymorphism is associated with the age of onset in schizophrenia. AB - OBJECTIVES: The NOTCH4 gene has a promoter polymor-phism at position -25, which leads to the three genotypes TT, CT and CC. These have been suggested to present a novel independent genetic risk factor for schizophrenia. We conducted a prospective case-control study to explore the impact of NOTCH4 T-25C polymorphism on the factors associated with schizophrenia. METHOD: NOTCH4 gene promoter T-25C polymorphism was determined by polymerase chain reaction among 94 patients with schizophrenia and 94 healthy age-matched and sex-matched blood donors. RESULTS: The T allele was highly associated with an earlier age of onset in male patients of schizophrenia (Kaplan-Meier log-rank test P<0.0001). Moreover, the male patients carrying the T allele were born significantly more often in June November compared with other months of the year [odds ratio=3.92 (95% confidence interval=1.025-15.018), P=0.046]. No association was determined, however, between the NOTCH4 gene polymorphism under study and schizophrenia. CONCLUSION: The NOTCH4 T-25C polymorphism has an important effect on the age of onset in schizophrenia and thus may be related to an early pathogenesis of schizophrenia in young patients. Alternatively, these findings may represent a significant genetic marker for managing subgroups and etiological clues in schizophrenia. PMID- 12782961 TI - Analysis of polymorphisms in the alpha-subunit of the olfactory G-protein Golf in lithium-treated bipolar patients. AB - OBJECTIVE: This study examines the alpha-subunit of the olfactory G-protein (G(olf)) as a possible candidate gene for bipolar disorder. The alpha-subunit of the G(olf) gene maps to a region on chromosome 18p that has been implicated in several linkage studies as a potential site of a bipolar disorder susceptibility loci. METHODS: We investigated whether two polymorphisms in the alpha-subunit of the G(olf) gene (A-->G in intron 3 and T-->G in intron 10) are associated with bipolar disorder in a sample of 149 bipolar patients under lithium treatment compared with 139 healthy controls using haplotype analysis. RESULTS: There was no evidence for an association between the investigated polymorphisms in the G(olf) gene and bipolar disorders, as well as to response to lithium treatment or common side effects, like hand tremor, weight gain and cognitive dysfunction. CONCLUSION: The results of the present study do not support the hypothesis that the G(olf) gene is a major susceptibility factor for bipolar disorders. PMID- 12782962 TI - Positive associations of polymorphisms in the metabotropic glutamate receptor type 3 gene (GRM3) with schizophrenia. AB - OBJECTIVES: Glutamatergic dysfunction is one of the major hypotheses of schizophrenia pathophysiology. We have been conducting systematic studies on the association between glutamate receptors and schizophrenia. We focused on the metabotropic glutamate receptor type 3 gene (GRM3) as a candidate for schizophrenia susceptibility. METHODS: We genotyped Japanese schizophrenics (n=100) and controls (n=100) for six single nucleotide polymorphisms (SNPs) located in the GRM3 region at intervals of approximately 50 kb. Statistical differences in genotype, allele and haplotype frequencies between cases and controls were evaluated by the chi2 test and Fisher's exact probability test at a significance level of 0.05. Haplotype frequencies were estimated by the EM algorithm. RESULTS: A case-control association study identified a significant difference in allele frequency distribution of a SNP, rs1468412, between schizophrenics and controls (P=0.011). We also observed significant differences in haplotype frequencies estimated from SNP frequencies between schizophrenics and controls. The haplotype constructed from three SNPs, including rs1468412, showed a significant association with schizophrenia (P=8.30 x 10-4). CONCLUSIONS: Our data indicate that at least one susceptibility locus for schizophrenia is situated within or very close to the GRM3 region in the Japanese patients. PMID- 12782963 TI - Genome scan of pedigrees multiply affected with bipolar disorder provides further support for the presence of a susceptibility locus on chromosome 12q23-q24, and suggests the presence of additional loci on 1p and 1q. AB - OBJECTIVE: To localize genes conferring susceptibility to bipolar affective disorder. METHODS: Seven families were selected on the basis of containing multiple cases of bipolar affective disorder present in three or more generations, an absence of schizophrenia and unilineal transmission. DNA samples from these families were genotyped with 365 microsatellite markers spaced at approximately 10 cM intervals across the whole genome. All markers were subjected to initial two-point and three-point analyses using LOD score and model-free analysis. All regions producing a result significant at P<0.01 were then subjected to four-point LOD score analysis under the assumption of heterogeneity. RESULTSA four-point LOD score of 2.8 was obtained using a dominant model and including unipolar cases as affected in the region of D12S342. Four-point LOD scores of 2 were obtained around D1S243, D1S251 and D3S1265. The positive results around D1S243 were accounted for by a LOD score of 3.1 occurring in a single pedigree. CONCLUSIONS: Since there has been previous strong support for linkage to the region of 12q23-q24 around D12S342, it now seems very probable that it does indeed contain a gene influencing susceptibility to bipolar affective disorder. Some evidence for linkage in the region of 1q near to D1S251 has been reported in one previous study. It therefore seems that this region of 1q and the region of 1p close to D1S243 may also harbour susceptibility genes. PMID- 12782964 TI - Joint analysis of candidate genes related to Alzheimer's disease in a Spanish population. AB - BACKGROUND: Besides the repeated association between the epsilon 4 allele of the apolipoprotein E and Alzheimer's disease, several candidate genes have been analysed with inconsistent results. Most of these studies have examined only one or two polymorphisms in a defined population, which provides a lack of a general view of the Alzheimer's disease genetic component. OBJECTIVE: To overcome this limitation, nine polymorphisms in seven different candidate genes (A2M, ACT, APOE, APP, BH, HSP70-2, and IL1-A) were genotyped. METHODS: The sample comprised 112 Alzheimer's disease patients and 89 controls from Spain. Since haplotype reconstruction may add power to association studies, we also tested for linkage disequilibrium within the A2M and APOE genes. RESULTS: Except for the APOE gene, allele and genotype frequencies were not different between cases and controls, even when stratifying for the APOE genotype. CONCLUSION: The present results suggest that future association studies should be performed using a battery of polymorphisms in different and new candidate genes, taking into account the linkage disequilibrium in the region. PMID- 12782965 TI - Schizophrenia, psychotic illness and other psychiatric symptoms in families with autosomal dominant nocturnal frontal lobe epilepsy caused by different mutations. AB - OBJECTIVES: Autosomal dominant nocturnal frontal lobe epilepsy (ADNFLE) is characterized by a strong family history of epileptic seizures, which predominantly occur during sleep. ADNFLE has been associated with mutations in two genes coding for the nicotinic acetylcholine receptor (CHRNA4 and CHRNB2). Thus far, three different mutations have been detected in the CHRNA4 gene, and two in the CHRNB2 gene. The aim of this study was to compare the frequency of psychiatric disorders in two ADNFLE families with different CHRNA4 mutations (776ins3 and Ser248Phe). METHODS: Information was gathered from hospital charts and therapists, and the family members were assessed by clinical interviews and structured clinical interviews. RESULTS: Of the 10 individuals diagnosed with epilepsy in the CHRNA4-776ins3 family, at least four had been in contact with psychiatric services. One individual had schizophrenia, while another family member had experienced at least two severe psychotic episodes, and had been taking antipsychotic medications for years. The third family member had been hospitalized at least three times for psychiatric problems. The fourth family member needs help with activities of daily living due to incapacitating apathy, although she does not have a psychiatric diagnosis. Such accumulation of psychiatric problems was not seen in the family with the Ser248Phe mutation. CONCLUSION: These findings suggest that there may be an association between the 776ins3 mutation and the psychiatric symptoms, a hypothesis that needs further testing. PMID- 12782966 TI - Mutational analysis of two positional candidate susceptibility genes for bipolar disorder on chromosome 12q23-q24: phenylalanine hydroxylase and human LIM homeobox LHX5. AB - OBJECTIVES: In the search for chromosome 12 genes potentially involved in the pathogenesis of bipolar disorder we will screen Phenylalanine hydroxylase and human LIM-homeobox LHX5 genes for sequence variants, both of which have been suggested as candidate genes. The genes lie on chromosome 12q23-24, near the Darier's disease gene, ATP2A2. We have previously reported two families in which the pattern of segregation of illness is consistent with genetic linkage between this chromosomal region and a putative highly penetrant autosomal dominant major affective disorder locus (pedigree 324, maximum LOD=2.1; pedigree 5501, maximum LOD=3.6). METHODS: We screened the coding and intronic flanking regions of the phenylalanine hydroxylase and LHX5 genes for sequence variation by denaturing high-performance liquid chromatography in individuals from the pedigrees. RESULTS: In total, nine single nucleotide polymorphisms and one 6 base pair deletion were identified. CONCLUSION: Our studies allowed us to conclude that none of these variants act as a highly penetrant autosomal dominant susceptibility locus for mood disorder in our families. PMID- 12782967 TI - Genetic variation in the seven-pass transmembrane cadherin CELSR1: lack of association with schizophrenia. AB - OBJECTIVES: Cadherins play a critical role in morphogenesis and maintenance of neuronal connections in the adult brain. We examined the gene encoding a member of the non-classic seven-pass transmembrane cadherins, CELSR1 for association with schizophrenia. It maps to chromosome 22q13.31, a region in which evidence for linkage to schizophrenia has been reported. The gene has an unusually large first exon of 3544 nucleotides, which encodes the signal peptide and all nine ectodomains in the protein. METHODS: We screened this exon in 24 schizophrenic patients using denaturing high-performance liquid chromatography followed by sequencing. Genotyping of amino-acid changes was performed with primer extension on a sample of 244 Bulgarian schizophrenic patients from 233 families and all their parents, as well as 180 schizophrenic patients from the UK and 157 controls. RESULTS: Three amino-acid changes were identified and shown to be in complete linkage disequilibrium: L556 V, S664W and R1126C. There was no preferential transmission of alleles from heterozygous parents to affected offspring. In the UK population the rare alleles were even more common in controls, and this difference almost reached statistical significance for R1126C (chi2=3.63, P=0.057). CONCLUSIONS: We conclude that variations in the nine ectodomains of CELSR1 do not increase susceptibility to schizophrenia. PMID- 12782968 TI - No evidence of association of two 5HT transporter gene polymorphisms and attention deficit hyperactivity disorder. AB - OBJECTIVES: Three studies to date have found evidence (or a trend for evidence) of linkage and association between the long allele of the 44 base pair repeat insertion/deletion 5-HTT functional polymorphism (5-HTTLPR) and attention deficit hyperactivity disorder (ADHD). In an attempt to replicate these findings, we examined this polymorphism and a variable number tandem repeat in the second intron of 5-HTT for association with ADHD. METHODS: One hundred and fifty children who met diagnostic criteria for ADHD and their parents (where available) were genotyped for these polymorphisms. Analysis was undertaken using the transmission disequilibrium test and haplotype analysis, as well as case-control comparisons using a control group of 121 individuals. RESULTS: No association between either the 5-HTTLPR or the variable number tandem repeat (VNTR) in ADHD was found (extended transmission disequilibrium test; P=0.37 and P=0.62, respectively). Haplotype analysis was also non-significant. Further analysis revealed no evidence of association in the subgroups of those without conduct disorder and in medication non-responders. CONCLUSIONS: Failure to replicate findings from previous studies may be due to a lack of statistical power. However, given recent findings by Kent et al. (2002) of association with another polymorphism in the 5HTT gene, we hypothesise that previous positive findings may have arisen by the LPR and VNTR being in linkage disequilibrium with the true susceptibility polymorphism. PMID- 12782969 TI - Cytochrome P450 2D6 polymorphism and character traits. AB - OBJECTIVE: It has been suggested that cytochrome P450 2D6 (CYP2D6) is involved in dopamine metabolism within the brain. The dopamine system is suggested to play a role in determining normal character. The purpose of this study was to examine whether character traits are dependent on cytochrome P450 2D6 activity. METHODS: We investigated the association between temperament and CYP2D6 gene polymorphism. The subjects were all Japanese and the polymorphism genotyped in the present study was CYP2D6*10. Character traits were assessed using the Temperament and Character Inventory. RESULTS: There was no overall or specific association between personality traits and the CYP2D6*10 allele and genotype frequencies. CONCLUSIONS: The present results do not support the hypothesis that CYP2D6 activity affects temperament and character. PMID- 12782970 TI - Quantitation of X-Y homologous genes in patients with schizophrenia by multiplex polymerase chain reaction. AB - OBJECTIVES: The genetic basis of schizophrenia is obscure. In an XX male patient with schizophrenia we previously showed that one X;Y translocation breakpoint was in pseudoautosomal region 1 (PAR1) with the effect that the proximal segment of PAR1 from the PAR1 boundary to acetylserotonin N-methyl transferase (ASMT) distally was triplicated in this patient. This study determined whether dosage imbalances of X-Y homologous regions in general are associated with schizophrenia. METHODS: A multiplex semi-quantitative polymerase chain reaction assay was developed to quantify MIC2 gene as a representative of PAR1 and compare it with the SYBL1 gene which maps in pseudoautosomal region 2 (PAR2) and protocadherin XY (PCDHXY), located at Xq21.3. Each of these three loci was co amplified with the autosomal gene MSX2 using Cy5-labelled primers and the products separated by electrophoresis in polyacrylamide gels. Results were expressed as ratios of peak area of the target gene to MSX2 which served as an internal dosage control. RESULTS: Using genomes with sex chromosome aneuploidies, the method was found sensitive enough to detect a two-fold difference in gene copy number. We confirmed the MIC2 triplication in the XX male patient but found no significant difference in gene dosage of MIC2, PCDHXY and SYBL1 in a panel of 17 patients with schizophrenia compared to controls. CONCLUSIONS: No evidence was obtained for gene dosage imbalances in MIC2, PCDHXY and SYBL1 in patients with schizophrenia. PMID- 12782971 TI - Multicentre Italian family-based association study on tyrosine hydroxylase, catechol-O-methyl transferase and Wolfram syndrome 1 polymorphisms in mood disorders. AB - OBJECTIVE: The aim of the present study was to investigate tyrosine hydroxylase, catechol-O-methyl transferase and Wolfram syndrome 1 genes in mood disorders using a family-based association approach. METHODS: The sample included 134 nuclear mood disorder families, with subjects affected by bipolar disorder (n=103) or major depressive disorder (n=58). All subjects were genotyped using polymerase chain reaction techniques. RESULTS: No significant transmission disequilibrium was found in the overall sample for any polymorphism. Analysis considering bipolar subjects only, or psychopathology traits as affection status did not influence the observed results. CONCLUSIONS: The study could not support the involvement of tyrosine hydroxylase, catechol-O-methyl transferase and Wolfram syndrome 1 polymorphisms in mood disorders. PMID- 12782972 TI - Impulsiveness as the intermediate link between the dopamine receptor D2 gene and alcohol dependence. AB - OBJECTIVES: Reinforcement and reward processes have been proposed as being an intermediate link between the risk for alcohol dependence and the gene coding for the dopamine receptor D2 (DRD2). This hypothesis remains open to speculation, and personality traits such as impulsiveness, a core dimension in addictive disorders, should also be taken into account. For instance, recent evidence in rats showed that DRD2 antagonists might increase impulsivity in decreasing the value of delayed rewards. METHODS: Considering the pro-impulsiveness role of ethanol observed in clinical practice and epidemiological studies, we analysed the Barratt impulsiveness scores in a sample of 92 alcohol-dependent French patients (57 men and 35 women), according to the TaqI A polymorphism of the DRD2 gene. RESULTS: A2/A2 and A1/A2 genotypes were significantly associated with a higher global impulsiveness than A1/A1 genotype (P=0.02 and P=0.03, respectively). CONCLUSIONS: We propose that reward-related impulsiveness may constitute a risk factor for alcohol dependence, and that this core temperament could be partly mediated by the DRD2 gene. PMID- 12782973 TI - Efficacy of cervical endplay assessment as an indicator for spinal manipulation. AB - STUDY DESIGN: Double-blind, randomized, placebo-controlled trial. OBJECTIVES: To evaluate the effect of manual endplay assessment on neck pain and stiffness outcomes in neck pain patients receiving spinal manipulation. SUMMARY OF THE BACKGROUND DATA: There have been no studies on the efficacy of palpation used as an indicator for manipulation in the management of back and neck pain. METHODS: Neck pain patients (n = 104) were randomly assigned to two groups. The study group received manipulation targeted to individual cervical vertebrae according to endplay restriction noted by the examining clinician. The control group received manipulation determined by sham, computer-generated examination findings; endplay examination was ignored and served as a placebo assessment. Treatment was rendered on a single occasion by a chiropractor. Outcomes were neck pain and stiffness assessed before and after manipulation and at least 5 hours following treatment. RESULTS: The study and control groups showed clinically important improvement in neck pain and stiffness. However, there were no clinically important or statistically significant differences between the study and control groups in terms of pain or stiffness outcomes. Findings were robust across patient, complaint, and treatment characteristics. CONCLUSIONS: Endplay assessment in and of itself did not contribute to the same-day pain and stiffness relief observed in neck pain patients receiving spinal manipulation. The impact on a longer course of treatment remains to be investigated. The data suggest that pain modulation may not be limited to mechanisms associated with manipulation of putative motion restrictions. PMID- 12782975 TI - Effects of a cervical compression plate on graft forces in an anterior cervical discectomy model. AB - STUDY DESIGN: An ex vivo biomechanical study using an instrumented distractor and load cells in a cadaveric multilevel discectomy construct was conducted. OBJECTIVE: To demonstrate that a dynamic cervical plate can be used to increase compressive load on interbody grafts in a multilevel discectomy specimen. SUMMARY OF BACKGROUND DATA: Cervical plating is used to decrease pseudarthrosis, graft extrusion, and graft subsidence in multilevel anterior discectomy procedures. Plating may shield a graft as it resorbs and may reverse normal loading mechanics. METHODS: Preoperative disc height was measured in five cadaveric spines. A three-level discectomy was performed. The disc spaces were opened with a distractor instrumented with strain gauges to allow the introduction of spacers fixed rigidly to subminiature load cells. Distraction was removed, and immediate compressive forces were measured by the load cells. An external compressor was applied followed by a cervical plate. The specimen then was placed in a loading frame, and final compressive forces were measured. RESULTS: A mean 116.5 N distractive force was required to insert grafts into all three levels. No significant relation between preoperative disc height and distractive or compressive forces was noted. Release of the distractor yielded an immediate compressive load on each graft. The compressor significantly increased graft compression. After plate application, the external compressor was removed. Graft compression did not significantly decrease. In the loading frame, an increase in compressive load was noted. CONCLUSIONS: Preoperative disc height was not related to the compressive force on the graft. Compressive force can be increased and maintained with a dynamic plating system. PMID- 12782976 TI - Effects of slip severity and loading directions on the stability of isthmic spondylolisthesis: a finite element model study. AB - STUDY DESIGN: Using a validated finite element model, the biomechanical effects of pars defect in a lumbar segment with and without different degrees of slip (up to 50% slip) were studied. OBJECTIVES: To study the effects of slip severity and loading parameters on the stability of the lytic and adjacent motion segments. Better knowledge of the biomechanics of spondylolisthesis may help formulate treatment strategies such as bracing or spinal implants. SUMMARY OF THE BACKGROUND DATA: Clinically, spondylolisthesis exists in varying grades of anterior slip, and the biomechanical stability of the motion segments at the lytic defect and adjacent level probably varies as well. In vitro studies of L4 L5 and L5-S1 isthmic spondylolisthesis slips have concluded that an L4-L5 pars defect is more unstable than an L5-S1 pars defect. Comparing the stability of lytic motion segments with different grades of spondylolisthesis is difficult to do experimentally and therefore has not been done. Further assessing the stresses in the bone and intervertebral discs at or adjacent to a lytic defect is also difficult to study experimentally, so no data are available. METHODS: A finite element model of L4-S1 was validated with and without a pars defect at L5. The model was then revised to represent different degrees of slip at L5, and six different moment loadings were applied. RESULTS: The current study showed larger decrease in stiffness with increasing percent slip. The decrease in disc stiffness and increase in disc stresses with increasing percent slip were larger at the level of spondylolisthesis as compared to the changes in the adjacent segment. Lateral bending moment and torsion load showed the largest decrease in stiffness due to slip. At 50% slip, the maximum increase in motion (as compared to motion in an intact segment) was seen under lateral bending moment load (about 55% at L4/L5 and 250% at L5/S1). Lateral bending also produced the largest increase in stresses due to 50% slip in the anulus and endplates (300% increase in anular stress and 190% increase in endplate stress) at L5/S1. CONCLUSIONS: The stiffness of a spondylolisthetic motion segment decreases as the slip increases. Lateral bending and torsion are moment directions causing the greatest resulting motions. PMID- 12782977 TI - Sitting with adjustable ischial and back supports: biomechanical changes. AB - STUDY DESIGN: The seat and back contact force, pressure distribution, lumbar lordosis, and low back muscle activities associated with a new seat design with adjustable ischial support and backrest were investigated using kinematic, kinetic, electromyographic, and radiographic measurements. OBJECTIVES: To investigate the biomechanical effects of adjusting ischial and backrest supports during sitting. SUMMARY OF THE BACKGROUND DATA: Sitting may induce posterior rotation of the pelvis, reduction of lumbar lordosis, and increases in muscle tension, disc pressure, and pressure on the ischium and coccyx, which may be associated with low back pain. A device that reduces the ischial load and maintains lumbar lordosis may help increase seating comfort and reduce low back pain. METHODS: Fifteen office workers with no known low back pain history were tested. Contact pressure distributions, reaction forces between the buttock thighs and seat and between the back and backrest, load carried by the seat pan and backrest, sacral inclination, lumbar lordosis, intervertebral space of lumbar spine, and muscular activity in stabilizing the trunk were measured for sitting with and without ischial support and with adjustable back support. RESULTS: When the ischial support was relieved, the center of the force on the seat and on the legs of the chair, and the peak center of pressure on the seat, were significantly (P < 0.002) shifted forward toward the thighs. The total contact area on the seat pan and on the backrest was significantly decreased and increased, respectively (P < 0.001). The sacral inclination, total and segmental lumbar lordosis, and lumbar spine disc height were significantly increased for sitting upright with backrest, with the lumbar curve close to that during standing. CONCLUSIONS: Sitting with reduced ischial support and fitted backrest to the lower spine altered the contact area, reduced peak pressure under the ischia, reduced muscular activity, maintained total and segmental lumbar lordosis, rotated the sacrum forward, and increased lumbar intervertebral disc heights, which could potentially reduce low back pain. PMID- 12782979 TI - Palsy of the C5 nerve root after midsagittal-splitting laminoplasty of the cervical spine. AB - STUDY DESIGN: The imaging characteristics of postoperative C5 nerve root palsy after midsagittal-splitting laminoplasty for cervical myelopathy, including those observed on plain radiography, computed tomography, and magnetic resonance imaging, were analyzed. OBJECTIVE: To investigate the imaging findings that predict occurrence of C5 nerve root palsy after midsagittal-splitting laminoplasty. SUMMARY OF BACKGROUND DATA: There have been several reports on imaging findings for postoperative nerve root palsy after open-door laminoplasty. However, there have been no detailed reports on imaging characteristics that predict the occurrence of nerve root palsy after midsagittal-splitting laminoplasty. METHODS: The study included 45 consecutive patients undergoing midsagittal-splitting laminoplasty with sufficient pre- and postoperative imaging examinations: 27 patients with cervical spondylotic myelopathy (CSM), 14 patients with ossification of the posterior longitudinal ligament (OPLL), and 4 patients with cervical disc herniation. Characteristics of pre- and postoperative plain radiographs, computed tomography scans, and magnetic resonance images were compared between the patients with and those without C5 nerve root palsy. RESULTS: Palsy of the C5 nerve root developed in 4 patients, and did not develop in 41 patients. Of the four patients with C5 nerve root palsy, one had CSM and the other three had OPLL. The incidence of C5 nerve root palsy involved 3 of 14 patients with OPLL patients (21.4%) and 1 of 31 patients without OPLL (3.2%) (P = 0.08). For both diseases, the patients with palsy tended to have increased postoperative cervical lordosis (P = 0.21). As for anterior compression on the spinal cord at C3, the P value for the comparison between the group with and the group without palsy was 0.07 for preoperative compression and 0.01 for postoperative compression. CONCLUSIONS: The preliminary data suggest that patients who have OPLL with marked anterior compression on spinal cord at C3 can be at risk for postoperative C5 nerve root palsy after midsagittal-splitting laminoplasty. Also, a postoperative increase in cervical lordosis may be the cause of postoperative nerve root palsy. PMID- 12782980 TI - Functional importance of degenerative spondylolisthesis in cervical spondylotic myelopathy in the elderly. AB - STUDY DESIGN: A correlation was studied between degenerative spondylolisthesis (DSL) of the cervical spine and spinal-evoked potentials intraoperatively recorded in elderly patients who had surgical treatment for cervical spondylotic myelopathy. OBJECTIVE: To investigate the functional importance of cervical DSL in elderly patients with cervical spondylotic myelopathy. SUMMARY OF BACKGROUND DATA: Cervical DSL has received insufficient attention in contrast to the lumbar DSL. The authors are unaware of any journal article in which this condition has been evaluated electrophysiologically. METHODS: This study investigated 47 patients with 68 DSL of 2 mm or more (3.1 +/- 0.9 mm; range, 2-6 mm) who underwent serial intervertebral recording of spinal-evoked potentials from either the intervertebral disc or the ligamentum flavum after epidural stimulation. RESULTS: All the study patients had unequivocal evidence of a focal conduction block, with the area of negative evoked potential peak reduced to less than 60% that of the immediately caudal level: 31 at C3-C4, 12 at C4-C5, and 1 each at C1 C2, C2-C3, C5-C6, and C6-C7. The site of conduction block matched the level of DSL in 30 patients, but not in 17 patients. The DSL accompanied by conduction block had significantly greater displacement with greater angular mobility than that without conduction block. CONCLUSIONS: A significant association between DSL and conduction block in the face of a relatively wide canal indicates the functional importance of DSL in elderly patients with cervical spondylotic myelopathy. In this age group, a high incidence of both DSL (81%) and focal conduction block (91%) at the upper cervical level (C3-C4 or C4-C5) is of clinical interest. PMID- 12782981 TI - One-stage posterior decompression-stabilization and trans-sacral interbody fusion after partial reduction for severe L5-S1 spondylolisthesis. AB - STUDY DESIGN: A retrospective clinical study was conducted. OBJECTIVES: To evaluate the clinical and radiologic outcomes of one-stage posterior decompression-stabilization after partial reduction and trans-sacral interbody fusion with a titanium cage for severe L5-S1 spondylolisthesis. SUMMARY OF THE BACKGROUND DATA: Trans-sacral interbody fusion for the management of severe L5-S1 spondylolisthesis with or without partial reduction and pedicular fixation has been previously described. The use of a trans-sacral titanium cage has not been previously reported. METHODS: Fifteen patients with severe L5-S1 spondylolisthesis were treated consecutively with posterior decompression, partial reduction, pedicular fixation, and posterior lumbar interbody fusion using a trans-sacral titanium cage. The mean age at the time of surgery was 22.4 years (range, 11-37 years). The mean follow-up period was 31.4 months (range, 12 58 months). Nine patients had severe back pain and six patients radicular pain. Three patients had a partial unilateral L5 motor deficit and two an L5 sensory deficit. Five patients had extremely tight hamstrings. The mean preoperative percentage of slipping was 69.3% (range, 53-91%). Patients were evaluated for complications and fusions, and outcomes were collected using the modified Scoliosis Research Society Outcomes Instrument. RESULTS: At follow-up, all patients, except one with major vascular complications, were extremely or reasonably satisfied with the surgery. All patients showed improvements in radiologic indexes and stable fusion at the final follow-up examination. CONCLUSIONS: Posterior decompression and partial reduction followed by circumferential stabilization performed in one stage combining pedicle fixation with trans-sacral titanium cage interbody fusion is an effective and safe technique for the management of severe spondylolisthesis. PMID- 12782982 TI - Risk factors for failure and complications of intradiscal electrothermal therapy: a pilot study. AB - STUDY DESIGN: A bi-institutional, retrospective clinical data analysis. OBJECTIVES: To determine risk factors for failure and complications of intradiscal electrothermal therapy, a treatment for discogenic back pain. SUMMARY OF THE BACKGROUND DATA: Intradiscal electrothermal therapy is a relatively new treatment for discogenic back pain. Though previous studies have shown it to be an effective treatment, there are few published studies examining complications and none examining risk factors for failure. METHODS: The authors treated 79 patients with discogenic back pain using intradiscal electrothermal therapy. Complications were assessed by patient report and, when indicated, further diagnostic testing. Success or failure was determined by visual analogue pain scores at 6-month follow-up. Variables examined for their relationship to failure and complications were age, sex, duration of pain, number of levels heated, smoking history, diabetes, obesity, leg pain, and previous back surgery. RESULTS: Forty-eight percent of patients reported more than 50% pain relief at their 6 month follow-up. There were eight complications (10%), most of which were self limited and transient. The only risk factor associated with intradiscal electrothermal therapy failure was obesity (P = 0.01). Whereas 54% of nonobese patients reported good pain relief at 6 months, only one out of 10 obese patients had successful intradiscal electrothermal therapy. The obese patients in our study were more likely to have a complication from intradiscal electrothermal therapy than they were to obtain pain relief. CONCLUSION: The only risk factor found to be associated with IDET outcome was obesity, which was a strong predictor of failure. Obesity should be considered a relative contraindication to performing IDET. PMID- 12782983 TI - Comparison of reliability between the Lenke and King classification systems for adolescent idiopathic scoliosis using radiographs that were not premeasured. AB - STUDY DESIGN: Multisurgeon comparison of two radiographic scoliosis curve classification systems was performed. OBJECTIVE: To determine the reliability of the King and Lenke classifications systems for adolescent idiopathic scoliosis using radiographs that had not been premeasured. SUMMARY OF BACKGROUND DATA: Recent studies introducing the new Lenke classification system for idiopathic scoliosis have reported reliability improved over that of the King classification system. This newer classification system evaluates three different parameters (curve type, lumbar modifier, and sagittal thoracic modifier) and then combines them. The reliability of both classification systems had been determined using radiographs in which all of the curves had been premeasured (recorded on the radiographs) before review by examiners. However, in a normal clinical situation, spine surgeons need to determine the Cobb angles independently, thus introducing another variable. METHODS: On two separate occasions, four orthopedic surgeons independently evaluated preoperative radiographs (standing posteroanterior, lateral, and two supine side-bending views) of 50 patients with adolescent idiopathic scoliosis. All measurements had been removed on every radiograph before each evaluation. The results were determined by calculating the average percentage of intraobserver and interobserver agreement. Reliability was quantified using kappa statistics. RESULTS: The King classification demonstrated good intraobserver and fair interobserver reliability. Intraobserver percentage of agreement averaged 83.5% (kappa coefficient, 0.81). Interobserver percentage of agreement averaged 68.0% (kappa coefficient, 0.61). All three parameters of the overall Lenke curve classification demonstrated fair reliability. Intraobserver percentage of agreement averaged 65.0% (kappa coefficient, 0.60). Interobserver percentage of agreement averaged 55.5% (kappa coefficient, 0.50). When the Lenke curve type was examined separately, intraobserver percentage of agreement averaged 81.5% (kappa coefficient, 0.76) and interobserver percentage of agreement averaged 71.5% (kappa coefficient, 0.64). The results for this variable (curve type) were similar to those for the King classification. For the Lenke lumbar modifier, the percentage of agreement and reliability were excellent. For the sagittal thoracic modifier, the percentage of agreement was good, but the kappa values were low because of an extreme imbalance in the grouping of hypokyphotic, normal, and hyperkyphotic spines. CONCLUSIONS: In this study, with each investigator performing the radiographic measurements, the King classification was found to be better than had been reported recently. The Lenke classification system for adolescent idiopathic scoliosis was found to be less reliable than previously reported when the radiographs were premeasured. This was particularly true when all three parameters of this new classification system were combined. This difference in reliability of the Lenke classification between studies can be attributed to the additional variable of determining the Cobb measurements on each of the unmarked radiographs. Although this new classification system has limitations with respect to interobserver and intraobserver reliability, for planning operative treatment, it offers a more comprehensive radiographic evaluation of patients with adolescent idiopathic scoliosis than previous systems. PMID- 12782985 TI - Three-dimensional quantitative segmental analysis of scoliosis corrected by the in situ contouring technique. AB - STUDY DESIGN: A three-dimensional analysis of right thoracic scoliosis before and after surgical correction was performed. OBJECTIVE: To compare the orientations of the vertebrae in apical and junctional zones before and after surgery. SUMMARY AND BACKGROUND DATA: Three-dimensional segmental analysis considering the lateral, sagittal, and axial (vertebral and intervertebral) rotations that can define apical and junctional zones has been described previously. Modern surgical techniques have been designed to correct scoliosis three-dimensionally. Therefore, the effect of the surgery must be evaluated in three dimensions. METHODS: The three-dimensional reconstruction of the spine and pelvis of 10 patients with right thoracic scoliosis was obtained by a stereoradiographic method before and after surgery. The orientations of all vertebrae were computed, and the apical and junctional zones were defined. The correction of these characteristic vertebrae obtained by the in situ contouring technique was evaluated. RESULTS: The mean three-dimensional pattern for the 10 patients was 20 degrees of vertebral axial rotation with 6 degrees of intervertebral extension in the thoracic apex and 30 degrees of vertebral lateral rotation with 10 degrees of intervertebral axial rotation in the junctional zones of the thoracic curve. The correction obtained by in situ contouring was 52% to 79% for all these three dimensional vertebral orientations. CONCLUSIONS: Three-dimensional reconstruction of the spine allows quantitative segmental analysis of the deformity. This could be used as a reliable tool to evaluate both the severity of the scoliosis and the effects of the surgical correction. PMID- 12782986 TI - Functional and radiographic outcomes after surgery for adult scoliosis using third-generation instrumentation techniques. AB - STUDY DESIGN: Retrospective radiograph and chart review of 28 patients with adult idiopathic scoliosis undergoing primary corrective surgery. Clinical and radiographic parameters were assessed before surgery, after surgery, and at a 2 year follow-up assessment. A self-perceived outcome questionnaire was administered to the study patients at a minimum 2-year follow-up assessment. OBJECTIVE: To assess patient outcomes after surgery for adult scoliosis using traditional radiographic parameters along with a self-perceived outcomes questionnaire. SUMMARY OF BACKGROUND DATA: The clinical and radiographic results and the outcomes for the surgical treatment of adult idiopathic scoliosis have not been established in the literature with respect to the use of modern third generation instrumentation techniques. Most studies reviewing the surgical treatment of adult idiopathic scoliosis look primarily at Harrington instrumentation techniques. METHODS: Records and radiographs were reviewed retrospectively for all the patients (n = 54) undergoing primary corrective surgery for adult idiopathic scoliosis between December 30, 1994 and December 30, 1997. Of the 54 patients reviewed, 28 (52%) met the following inclusion criteria: age exceeding 20 years, primary surgery, fusion above the sacrum, availability of medical records along with preoperative, postoperative, and 2-year follow-up radiographs. Additionally, a self-perceived outcomes questionnaire was administered to these patients at a minimum 2-year follow-up assessment. RESULTS: All the patients were women (28/28). The indications for surgery were pain and progression in 54% (15/28) and pain in 29% (8/28) of the patients. The average preoperative major curve measurement was 65 degrees (range, 38-98 degrees ). The average postoperative major curve measurement was 24 degrees (range, 5-59 degrees ), for a correction of 64%. The average follow-up curve measurement was 27 degrees (range, 3-60 degrees ), for a correction of 61%. Whereas 71% of the cases were anteroposterior, 29% were posterior only. There was one intraoperative complication among the 28 patients and four postoperative complications in 3 of the 28 patients. The self-perceived outcome questionnaires were available for 83% (23/28) of the patients. Definite or probable relief of symptoms was reported in 74%(17/23). Improved ability to sleep was reported in 61% (14/23), and ability to return to their usual job was reported in 57% (13/23). Satisfaction with the results of surgery was reported in 87% (20/23). CONCLUSIONS: Surgery for adult idiopathic scoliosis using third-generation instrumentation techniques provides significant clinical improvement, scoliosis correction, maintenance of sagittal alignment, and patient satisfaction, with an acceptable complication rate. PMID- 12782988 TI - Courses of cervical disc herniation causing myelopathy or radiculopathy: an analysis based on computed tomographic discograms. AB - STUDY DESIGN: The courses of protruded masses in cervical disc herniations were traced on preoperative computed tomography discograms of patients with myelopathy or radiculopathy. OBJECTIVE: To characterize the courses of protruded masses in cervical disc herniation. SUMMARY OF BACKGROUND DATA: No studies have been reported on the varied courses of protruded masses in cervical disc herniation. METHODS: This study investigated the preoperative CT discograms of 150 patients with myelopathy and 50 patients with radiculopathy who had undergone anterior cervical discectomy and fusion for disc herniation. The courses of herniations were traced from the penetration sites on the deep layer of the posterior longitudinal ligament through their locations in the spinal canal, and were divided into one median, two paramedian, and two lateral sections. RESULTS: Of the 150 discs in the patients with myelopathy, 87% had a median penetration and 13% had a paramedian one. No discs had a lateral penetration. It was found that 45% of the median penetrations led to median herniation through a straight course and 55% to paramedian herniation through an oblique course, and that 95% of the paramedian penetrations led to paramedian herniation through a straight course. Of the 50 discs in the patients with radiculopathy, 70% had a median penetration, 26% a paramedian penetration, and 4% a lateral penetration. Lateral penetration was observed only at C7-T1. All of the median penetrations led to the paramedian or lateral herniation, and 92% of the paramedian penetrations led to lateral herniation through oblique courses. CONCLUSIONS: In the cervical spine, most herniated masses penetrate the deep layer of the posterior longitudinal ligament in the middle, where the posterior intervertebral space is widest. Oblique courses to paramedian or lateral herniation are common. Only at C7-T1, where there are no Luschka joints, lateral penetration was observed. The narrow space of the Luschka joint may prevent fragments from penetrating laterally. Preoperative information from the CT discograms on the characteristic courses of the herniation may facilitate the complete removal of herniated mass in anterior decompressive surgery. PMID- 12782989 TI - Standing lateral radiographic positioning does not represent customary standing balance. AB - STUDY DESIGN: Normal cohort evaluation of the accuracy of existing methods for radiographic measurement of sagittal spinal balance. OBJECTIVES: To examine the validity and reliability of sagittal vertical axis measurements during a variety of standing positions commonly used while obtaining lateral thoracolumbar spine radiographs. SUMMARY OF THE BACKGROUND DATA: The sagittal vertical axis is a widely accepted radiographic measurement of global sagittal alignment of the spine. However, the sagittal vertical axis has not been measured in normal subjects while in functional positions because the arms must be elevated during acquisition of a lateral spinal radiograph. The purpose of this study was to quantify differences in sagittal vertical axis measurements between repeated functional positions and radiographic positions to identify the magnitude of the potential error in measuring the sagittal vertical axis radiographically. METHODS: Reflective markers were attached to 15 healthy female adolescents, overlying the spinous processes of C7 and S1. Marker positions were recorded by a motion capture system during simultaneous acquisition of a lateral radiograph. Sagittal vertical axis calculation, using marker data, was matched to sagittal vertical axis measurement obtained by radiograph using anthropometric corrections to limit radiation to a single exposure. Four standing positions with varying shoulder and knee flexion as well as overground walking were examined. The mean sagittal vertical axis for each standing position and during gait was compared using a repeated measures analysis of variance. Intratrial and intertrial repeatability of sagittal vertical axis measurements was also determined. RESULTS: The sagittal vertical axis was positive (C7 anterior to S1) for the functional positions (relaxed standing: 0.9 +/- 2.0 cm, and throughout gait: 4.5 +/- 2.0 cm), whereas shoulder flexion resulted in a negative sagittal vertical axis (-4.6 +/- 3.2) and posterior rotation of the pelvis. Adding knee flexion resulted in a slight relative shift in the sagittal vertical axis anteriorly. No differences were observed in intertrial and intratrial reliability for relaxed standing and standing with shoulder flexion alone. Increased variability was observed between repeated trials involving knee flexion. CONCLUSIONS: Measurement of the sagittal vertical axis on radiographs from commonly utilized standing positions (shoulders flexed) results in an sagittal vertical axis that is at least 3 to 4 cm more posterior than a sagittal vertical axis observed during a functional position. Subject repositioning resulted in an intertrial variability of at least 0.8 cm in sagittal vertical axis, while variation as the subject held each standing posture had little contribution to overall error of measurement. Of the analyzed positions, shoulder flexion (45 degrees ) alone was the best position for a lateral radiograph due to minimal compromise to repeatability of sagittal vertical axis measurement. However, none of the radiographic positions reproduced the spinal balance of the subject's functional standing posture. PMID- 12782990 TI - Vertebral artery loop formation: a frequent cause of cervicobrachial pain. AB - STUDY DESIGN: Patients with cervicobrachial pain were examined in detail with special attention to vertebral artery loop formation. OBJECTIVES: To determine the incidence, short-time natural course and response to a conservative approach to vertebral artery loop formation in a group of patients with cervicobrachial pain. SUMMARY OF THE BACKGROUND DATA: Vertebral artery loop formation is reported to be a rare cause of cervicobrachial neuralgia, discovered incidentally during the search for its cause. It can be congenital or acquired, occurring equally in both sexes. Surgical decompression has been the preferred method in the majority of reported cases, with favorable results. METHODS: One hundred seventy-three patients with cervicobrachial pain were examined in a period of 7 months using physical examination, radiography, and magnetic resonance imaging with or without angiography. RESULTS: Thirteen patients with a mean age of 43.9 +/- 13.5 years were diagnosed with vertebral artery loop formation. The most common level was C6 C7. Four patients presented with loop formation at two levels. None of the patients had symptoms attributable to intervertebral disc pathology. The complaints were in accordance with the level of the vascular pathology. Complete relief or decrease in pain was observed in all patients with the conservative approach. CONCLUSIONS: In patients with cervicobrachialgic symptoms and without established discopathy, during the examination of sagittal magnetic resonance images, vertebral artery loop formation should be kept in mind, and in suspected cases, the vertebral artery should be visualized using three-dimensional time of flight magnetic resonance angiography. Vertebral artery-nerve root relation should also be demonstrated using the multiplanar reformatting method from time of flight images. A conservative course of treatment has a favorable outcome. PMID- 12782991 TI - Physical impairment index: reliability, validity, and responsiveness in patients with acute low back pain. AB - STUDY DESIGN: Cohort study of patients with acute low back pain undergoing physical therapy. OBJECTIVES: Examine the reliability and validity of the Physical Impairment Index in a group of patients with acute low back pain and determine the responsiveness and minimum detectable change of the index and its component tests. SUMMARY OF THE BACKGROUND DATA: The Physical Impairment Index was originally described as a reliable and valid means of assessing physical impairment in patients with low back pain. The psychometric properties of the index have not been reported in patients with acute low back pain, nor has its responsiveness been examined. METHODS: Seventy-eight patients with acute (<3 weeks duration) low back pain participating in a clinical trial were assessed at baseline and after 4 weeks. Interrater reliability of the index was examined in a subgroup of 20 patients. Validity was examined through correlations with concurrent measures of pain, disability, and psychosocial variables. Changes in the index over 4 weeks were used to assess responsiveness and minimum detectable change. RESULTS: Interrater reliability of the index was high (intraclass correlation coefficient = 0.89), and its validity was generally supported by the pattern of correlations. The index was more responsive to change than any of its component tests but was less responsive than the Oswestry disability score. The minimum detectable change on the index was approximately 1 point. CONCLUSIONS: The Physical Impairment Index appears to be a reliable and valid measure of physical impairment for patients with acute low back pain and may be useful as an adjunct outcome measure for studies involving these patients. Further research on patients with chronic pain is needed before it can be advocated for outcomes research with this population. PMID- 12782992 TI - Prevalence and predictors of intense, chronic, and disabling neck and back pain in the UK general population. AB - STUDY DESIGN: Multiphase cross-sectional survey of musculoskeletal pain. OBJECTIVES: To estimate the prevalence of all reported and clinically significant spinal pain. To identify independent predictors of spinal pain. METHODS: A total of 5752 adults sampled from three general practice registers were mailed a screening questionnaire. Subjects who reported the spine as a predominant site of pain were sent a site-specific questionnaire (i.e., back or neck) that asked about severity. Prevalence estimates were calculated and extrapolated to the general population. Predictors of spinal pain were identified using logistic regression with comprehensive adjustment for confounders (including pain at other anatomic sites). RESULTS: The 1-month-period prevalence of all reported spinal pain was 29% (95% confidence interval 27-31%), of which about half was intense, half was chronic, 40% was disabling, and 20% was intense, disabling, and chronic. Most people with back (75%) or neck (89%) pain also reported pain at other sites. Age, female gender (neck pain only), high body mass index, living in an area of raised material deprivation, and south Asian ethnicity were significant predictors of spinal pain with disability. The association between body mass index and deprivation and neck pain was lost after adjustment for pain at other sites. However, even after full adjustment, obesity (OR, 1.7; 95% confidence interval, 1.1-2.5) and high deprivation (OR, 1.7; 95% confidence interval, 1.1 2.7) were predictors of back pain with disability. CONCLUSIONS: Adjustment for pain at other sites enabled assessment of whether observed associations were with spinal pain itself or with the other sites of pain. Obesity is an important independent predictor of back pain and its severity. This has implications for primary prevention. The prevalence of spinal pain with disability continues to rise into old age. This has implications for healthcare planning. PMID- 12782993 TI - Muscle flap salvage of spine wounds with soft tissue defects or infection. AB - STUDY DESIGN: This retrospective study was designed to analyze the results of 22 patients treated for postoperative soft tissue defects of the spine. OBJECTIVE: To demonstrate the utility of flaps in the salvage of spine wounds. SUMMARY OF BACKGROUND DATA: In the literature, the treatment of postoperative spine infections is with serial debridement, antibiotic irrigation catheters, drains, and occasional removal of spinal implants. Muscle flaps have received scant mention in the surgical literature for spine coverage. METHODS: Group 1 (n = 15) had postoperative wound infections or dehiscences. Group 2 (n = 7) had "prophylactic" flaps at the time of their initial spine surgery. The indications for "prophylactic" closure included multiple prior surgeries, prior infection, and previous radiation therapy. Group 1 was treated with drainage, dressing changes, and one-stage flap closure of their wounds. Sliding paraspinal muscle flaps were the flaps of choice. Group 2 was treated with a variety of closure techniques at the time of their initial surgery. RESULTS: The average defect size was 10 vertebral bodies long. Despite the large defect size, 19 of 20 surviving patients currently have healed wounds, and all the patients have maintained their instrumentation. Two patients died of causes unrelated to their wound problems. A Group 1 patient with complete loss of a superior gluteal artery flap was salvaged with a contralateral gluteus muscle flap. Another Group 1 patient has intermittent drainage from under a trapezius flap, which covers a cervical spine fusion. Four patients had minor wound complications. CONCLUSIONS: Flaps are a useful adjunct in the treatment of patients with complex spine wounds. Sliding paraspinal muscle flaps can effectively close wounds from the high cervical to the low lumbar area in one operative procedure. These patients can go on to successful spine fusion. PMID- 12782994 TI - Atlantoaxial rotatory fixation secondary to tuberculosis of occiput: a case report. AB - STUDY DESIGN: A case of nontraumatic rotatory fixation of the atlantoaxial joint associated with tuberculosis of the occipital bone in an adult. OBJECTIVES: To report a rare case of atlantoaxial rotatory subluxation associated with tuberculosis of the occipital bone in an adult and to discuss the mechanism of fixation. SUMMARY OF THE BACKGROUND DATA: Atlantoaxial rotatory fixation in adults is rare and has been reported due to variety of causes. To the authors' best knowledge no case has been reported secondary to tuberculosis of the skull bone. METHODS: A 20-year-old male presented with resistant torticollis with a duration of 5 months. RESULTS: The patient had type 1 atlantoaxial rotatory fixation secondary to tuberculosis of the occipital bone. The subluxation was partially reduced by conservative means, and healing of the occiput lesion was achieved. Thereafter, the patient had no restriction of cervical spine motion and had no reoccurrence of subluxation at a follow-up of one and a half years. CONCLUSIONS: Effusion in the atlantoaxial joint secondary to infection in the occiput due to close proximity with the joint led to the laxity of ligaments and contributed to the subluxation. PMID- 12782995 TI - Symptomatic hematoma of lumbar facet joint: joint apoplexy of the spine? AB - STUDY DESIGN: A case report. OBJECTIVES: To report and discuss a rare case of epidural hematoma that was considered to be formed as a result of idiopathic bleeding occurring at the facet joint (joint apoplexy). SUMMARY OF THE BACKGROUND DATA: Spontaneous spinal epidural hematoma is a relatively rare condition. According to a review article of 199 spontaneous spinal epidural hematomas in the last 2 decades, the majority of these conditions are thought to result from a rupture of the epidural vascular network. Recently, a hemorrhagic lumbar synovial cyst and a hematoma occurring from the ligamentum flavum were reported as rare types of epidural hematoma. METHODS: The authors describe the treatment and the clinical, radiologic, surgical, and pathologic findings in one patient with a rare epidural hematoma. RESULTS: Magnetic resonance imaging revealed that the extradural mass lesion was continuous with the right L4-5 facet joint; this was confirmed by surgery when the extradural hematoma was directly visualized. The joint cavity was also filled with the hematoma. There was no evidence of preceding cyst formation macroscopically or microscopically. The excised capsule of the left L4-5 facet joint revealed moderate hyperplasia of the synovium with an increased number of capillary vessels. CONCLUSIONS: This is the first reported case of radiculopathy considered to be a result of facet joint apoplexy in the absence of any preceding synovial cyst formation. The pathomechanism of the hemorrhage at the lumbar facet joint is unclear, but it is speculated that there could be an association with degenerative change of the facet joint. Surgical excision of this mass was considered to be the definitive treatment. PMID- 12782996 TI - Distant skip level discitis and vertebral osteomyelitis after caudal epidural injection: a case report of a rare complication of epidural injections. AB - STUDY DESIGN: A case report of distant discitis and vertebral osteomyelitis involving skip levels after caudal epidural steroid injection. OBJECTIVES: To report and investigate the occurrence of distant infective discitis and vertebral osteomyelitis involving skip levels after epidural injection. SUMMARY OF THE BACKGROUND DATA: Distant discitis and vertebral osteomyelitis is a serious but rare complication after epidural injection. A case involving skip levels and without the occurrence of epidural abscess formation has apparently not been previously reported in the literature. METHODS: An elderly woman presenting with clinical, radiologic, and magnetic resonance imaging evidence of spinal canal stenosis involving L3/4 and L4/5 levels and degenerative spondylolisthesis of the L4/5 level was given an epidural injection of steroids and lignocaine via the caudal route. A month later, she presented with worsened low back pain, elevated serum acute phase reactants, and plain radiographic evidence of L4/5 infective discitis. Magnetic resonance imaging and microbiologic examination of computed tomographically guided biopsy specimens confirmed infective discitis involving L2/3 and L4/5 intervertebral levels, together with adjacent vertebral osteomyelitis. RESULTS: The patient was successfully treated with antibiotics targeted at Pseudomonas aeruginosa, which was isolated in the culture of the biopsy specimens. Follow-up improvements in the clinical condition, serum acute phase reactants levels, radiographs, and magnetic resonance imaging were noted. CONCLUSIONS: Distant discitis and vertebral osteomyelitis involving skip levels and without the occurrence of epidural abscess formation is a serious but rare complication after epidural injection. PMID- 12782997 TI - Percutaneous CT-guided puncture and steroid injection for the treatment of lumbar discal cyst: a case report. AB - STUDY DESIGN: A case report of discal cyst treated by percutaneous CT-guided puncture and steroid injection. OBJECTIVES: To present the efficacy of percutaneous CT-guided puncture and steroid injection for the treatment of this disorder. SUMMARY OF THE BACKGROUND DATA: Surgical removal of the cyst is usually performed for the treatment of this disorder. However, it was recently reported that direct CT-guided puncture and steroid injection was useful for the treatment of intraspinal synovial cyst. METHODS: A 37-year-old man experienced low back pain and right lower extremity pain with sensory abnormality over the right leg and foot. MRI revealed an extradural spherical mass with low signal intensity on T1-weighted imaging and high signal intensity on T2-weighted imaging behind the L5 vertebral body. On discography, contrast medium rapidly flowed into the cyst through a thin channel from the neighboring disc cavity. Based on these findings, a diagnosis of discal cyst was made. Percutaneous CT-guided puncture and steroid injection were performed. RESULTS: Low back pain, right extremity pain, and numbness disappeared immediately after the procedure. On MRI examination, the cyst gradually diminished in size after the procedure. CONCLUSION: CT-guided percutaneous puncture and steroid injection may be a useful procedure for the treatment of discal cyst. PMID- 12782999 TI - Aesthetic surgery 101: resident education in aesthetic surgery, the MGH experience. PMID- 12782998 TI - Chronic inflammatory granuloma mimics clinical manifestations of lumbar spinal stenosis after acupuncture: a case report. AB - STUDY DESIGN: A case report and review of the literature. OBJECTIVES: To present a case of chronic inflammatory epidural granuloma formed after acupuncture. SUMMARY OF THE BACKGROUND DATA: A number of cases of complications resulting from acupuncture have been reported, including acute infection, hemorrhage, and direct injury to internal organs or neural tissues. However, to the best of our knowledge, there has been no report of epidural granuloma formed following acupuncture and mimicking clinical manifestations of lumbar stenosis. METHODS: A 68-year-old woman suffered from low back pain and sciatica aggravated by acupuncture. We reviewed her medical record, imaging studies, microscopic findings of the mass, and related literature. RESULTS: Microscopic examination revealed the mass as a chronic inflammatory granuloma. From her previous history and imaging study, the mass, which compressed the lumbar forth nerve and dural sac,was highly suspected to have been formed after acupuncture. Surgical decompression and excision of the epidural mass relieved her symptoms. CONCLUSION: Chronic inflammatory granuloma may be formed as a complication of acupuncture. Under such circumstances, surgical excision of the mass may be an effective way of relieving the symptoms. PMID- 12783000 TI - TRAM flap breast reconstruction for patients with advanced breast disease. AB - Transverse rectus abdominis musculocutaneous (TRAM) breast reconstruction in patients with advanced breast cancer is controversial. Management of these patients is often complex and consists of surgical extirpation, postoperative radiation, chemotherapy, and in some cases bone marrow transplantation. Few studies have attempted to examine patient long-term survival and overall satisfaction with the surgical procedure. This study examines one center's experience with patients undergoing breast reconstruction for stage III and stage IV breast carcinoma. A retrospective review was performed of all patients undergoing TRAM reconstruction with stage III or IV breast cancer. Surviving patients and family members were contacted for follow-up. Patients were asked to grade their satisfaction with the reconstructive procedure on a 5-point scale (5 points, extremely satisfied; 1 point, extremely dissatisfied). Postoperative complications and time to return to work were also recorded. During a 10-year period (1991-2000) 21 women underwent TRAM reconstruction for advanced breast cancer. Twenty patients had stage III disease and 1 patient had stage IV disease. Mean patient age was 49 years. A total of 26 TRAM flaps were performed; 5 patients had bilateral procedures. Of the 26 TRAM flaps, 17 were immediate and 9 were delayed, and 20 were free and 6 were pedicled. Follow-up averaged 6.5 years (range, 2-10 years). Postoperative complications occurred in 7 patients and included fat necrosis (N = 3), hematoma (N = 2), cellulitis (N = 1), delayed donor site healing (N = 2), and seroma (N = 1). There were no flap losses. Patients were able to return to normal activities or work at an average of 10.6 weeks. Eleven patients developed recurrent disease. Nine patients (43%) succumbed to their disease during the follow-up period. In these patients the average interval between TRAM reconstruction and death was 3.7 years (range, 1-6.5 years). Eleven patients or surviving family members participated in the patient satisfaction survey. The average satisfaction grade was 4.6 points. All patients would repeat the TRAM reconstruction again. Patients with advanced breast cancer can be considered appropriate candidates for TRAM reconstruction. The results of this study indicate that patients with advanced breast cancer do not have an increased rate of postoperative complications, and they recover within a reasonable time from their surgical procedure despite adjuvant radiation and chemotherapy. Furthermore, the majority of patients are satisfied with their reconstructed breast and postoperative course, and would choose this reconstructive option again. PMID- 12783001 TI - Breast reconstruction in the elderly: preserving excellent quality of life. AB - There is a paucity of information in the literature that focuses on quality-of life issues after mastectomy with breast reconstruction in elderly women. The purpose of this study was to review the authors' experience with breast reconstruction after mastectomy in women older than 65 years of age. Emphasis was placed on the types of reconstructions, outcomes, and evaluation of issues related to quality of life. Between July 1997 and July 2001, 316 consecutive women (400 reconstructions) with breast cancer underwent mastectomy with reconstruction at the authors' institution. Of these women, 24 (28 reconstructions) were at least 65 years old. Mean patient age was 69.3 years (range, 65-77 years). Methods of breast reconstruction included two-stage implant reconstruction (50%), free tissue transfer (transverse rectus abdominis musculocutaneous or deep inferior epigastric perforator flap, 25%), pedicle transverse rectus abdominis musculocutaneous flap (14%), or a latissimus dorsi musculocutaneous flap (11%). Outcomes were assessed with the use of a self reported questionnaire (SF-36) addressing health-related quality of life, body image, and physical functioning. With respect to overall quality-of-life issues after reconstruction, older patients with breast reconstruction scored higher (better outcomes) than age-matched general population patients and previously reported mastectomy-only patients (>55 years) in all surveyed areas. Specifically, study patients reported dramatic increases (better outcomes) in the subscales that are strongly influenced by one's mental health. However, when compared with prior data for younger patients undergoing mastectomy and reconstruction, the older patients scored lower (worse outcomes) in the areas related to physical function. The older patients maintained superior scores (better outcomes) over the younger patients in the subscales influenced by one's mental health. PMID- 12783003 TI - Innovations to the vertical reduction mammaplasty: making the transition. AB - The vertical reduction mammaplasty can be challenging to learn. In addition, first attempts to perform the vertical reduction mammaplasty can lead to inconsistent aesthetic results. The authors describe their transition from a traditional inverted-T reduction mammaplasty to a modified vertical reduction mammaplasty based on a technique described by Elizabeth Hall-Findlay. In their early cases using the Hall-Findlay technique, they noted several aesthetic complications. These problems included a persistent vertical dog-ear deformity at the nadir of the incision, a teardrop deformity of the nipple-areola complex, lateral deviation of the nipple, and lateral axillary fullness. They developed several modifications to the Hall-Findlay technique to correct the aesthetic deficiencies and to simplify further the vertical reduction method. The authors think their innovations facilitate the transition from a traditional inverted-T breast reduction to a successful vertical reduction mammaplasty technique. PMID- 12783005 TI - Tissue expansion after inverted-T mammaplasty: can it be performed successfully? AB - Breast cancer patients will occasionally need to undergo mastectomy after previous reduction mammaplasty or mastopexy. The presence of the "inverted-T" scar presents a unique reconstructive dilemma: Do mastectomy flaps that are traversed by surgical scars still allow for adequate tissue expansion and a good aesthetic result? The objective of this study was to evaluate the authors' experience with tissue expansion/implant reconstruction in patients with an inverted-T scar. All patients who underwent tissue expansion and implant reconstruction after inverted-T mammaplasty were reviewed retrospectively to determine aesthetic results, patient satisfaction, and complication rates associated with this reconstructive technique. During a 6-year period (1995 2001), 11 patients (12 breasts) underwent breast reconstruction with tissue expansion and implant placement after either reduction mammaplasty (N = 8) or mastopexy (N = 4). All patients reached target expansion volumes. After successful tissue expansion, exchange to either saline (N = 7) or silicone (N = 5) implants was performed. The mean follow-up period after implant exchange was 18.5 months (range, 2-72 months). Complications were minimal and included partial flap necrosis (N = 1). Aesthetic appearance and symmetry were judged to be good or excellent in the majority of patients. Overall patient satisfaction was high. Tissue expansion with implant exchange is an effective reconstructive technique for mastectomy defects after previous inverted-T mammaplasty. In this series, good to excellent aesthetic results were achieved in the majority of patients with minimal associated complications. PMID- 12783007 TI - The cleft lateral lip element: do traditional markings result in secondary deformities? AB - In cleft surgery, two methods have traditionally been used to mark the height of cupid's bow on the lateral lip element. One technique measures the distance from the oral commissure to the height of cupid's bow on the noncleft side, and transposes this distance onto the cleft-side lateral lip element. The second technique marks the height of cupid's bow on the cleft-side lateral lip element where the white roll disappears. The authors believe these techniques may result in deformities of residual cleft tissue in the repair. Marking the height of cupid's bow on the cleft-side lateral lip element, just before the attenuation of lip fullness, can prevent this deformity. A retrospective study yielded a series of 17 patients with secondary deformities of residual cleft tissue in their repair. The method used to mark the lateral lip element was determined by chart review. Patients then underwent secondary surgery with excision of residual cleft tissue, and repair using the initial technique. A random group of primary cleft patients, repaired using the authors' technique for marking the lateral lip element, was likewise evaluated for the presence of residual cleft tissue in the repair. Of the 17 cases of secondary deformities, 14 were unilateral and 3 were bilateral. Among the unilateral cases, seven were repaired with a triangular flap and seven by rotation advancement. The bilateral cases were repaired using the modified Millard technique. The lateral lip element was marked using cessation of the white roll in 8 patients, and the commissure to the height the of cupid's bow in 2 patients, whereas in 7 patients the method was unreported. Using the authors' technique, both "controls" repaired primarily and cases repaired secondarily resulted in no redundant cleft tissue. Average follow-up was 11 months (range, 1-41 months). The authors think that traditional markings for establishing the height of cupid's bow on the cleft lateral lip element may result in residual cleft tissue in the repair. This deformity can be prevented by marking the height of cupid's bow on the cleft lateral lip element just before the attenuation of lip fullness. PMID- 12783009 TI - Sentinel lymph node biopsy for the T1 (thin) melanoma: is it necessary? AB - The use of sentinel lymph node biopsy for the T1 melanoma is controversial. Recent reports have demonstrated that certain T1 melanomas are at increased risk for early regional metastases and late recurrence when compared with all thin melanomas. The purpose of this study was to review the authors' experience with wide excision and sentinel lymph node biopsy for certain patients with T1 melanoma. A retrospective analysis of 34 patients with T1 melanoma was completed over a 3-year period. Indications for sentinel lymph node biopsy included a Breslow thickness of less than or equal to 1 mm a Clark level of III or IV tumor ulceration, or tumor regression. Twenty-four patients met these criteria (13 men and 11 women). Mean age was 47.6 years (range, 23-88 years). Mean tumor thickness for all patients was 0.69 mm (range, 0.3-1.0 mm), 0.61 mm for the Clark level III patients (N = 15), and 0.72 mm for the Clark level IV patients (N = 9). Tumor ulceration was present in 1 patient and histological regression was present in 2 patients. Regional lymph node metastases were confirmed histologically in 2 of 24 patients (8.3%) in whom the thickness of the melanoma was 0.9 mm and 1 mm. Both patients have died of metastatic melanoma. No recurrence has been demonstrated in the remaining 22 patients at the 2 to 5-year follow-up. Current indications for sentinel lymph node biopsy for patients with T1 melanoma include tumors associated with Clark level IV or V invasion, ulceration, regression, a positive deep margin on initial biopsy, or previous melanoma. Acral lentiginous melanoma associated with at least a Clark level III invasion warrant sentinel lymph node biopsy. Superficial spreading or nodular melanoma larger than 0.9 mm should include sentinel lymph node biopsy regardless of other associated histological factors. PMID- 12783011 TI - How accurate is frozen section for skin cancers? AB - Frozen section analysis has been used to increase the likelihood of complete excision of skin cancers and to minimize the risk for recurrence. The question of its accuracy has been addressed in many studies and this article adds data to the discussion. A retrospective study was performed of 60 consecutive cases in which frozen section diagnoses were compared with permanent sections. It was found that in 85% of the cases the frozen sections were accurate compared with the permanent sections, but in 13% of the total cases the margins were less than 1 mm. These results concur with the literature. The surgery and pathological examinations were performed in a community hospital in which there was no dermatopathologist. The authors review the limitations of frozen section diagnosis in skin cancers and show that frozen section may be necessary in select cases but not in most skin cancers. PMID- 12783012 TI - Biological alchemy: engineering bone and fat from fat-derived stem cells. AB - Adipose tissue contains a population of pluripotent stem cells capable of differentiating along multiple mesenchymal cell lineages. In this study the authors isolated these fat-derived stem cells successfully from Lewis rats and induced differentiation along adipogenic and osteogenic lineages in vitro and in vivo. Induction was stimulated by exposing stem cells to lineage-specific induction factors. Adipocyte-inducing media contained dexamethasone, insulin, and isobutyl-methylxanthine. Osteoblast inducing media contained dexamethasone, beta glycerophosphate, and ascorbic acid. Undifferentiated stem cells were maintained in minimal essential media alpha and fetal bovine serum. At 10 days, cells cultured in adipogenic media differentiated into adipocytes in vitro, as evidenced by positive Oil red O staining of lipid vacuoles. At 21 days, cells cultured in osteogenic media differentiated into osteoblasts in vitro as demonstrated by Alizarin red staining of a calcified extracellular matrix and immunohistochemical staining for osteocalcin. Differentiated cells were seeded at a density of 5 x 106 cells onto 15 x 15-mm polyglycolic acid grafts and implanted subcutaneously into three groups of Lewis rats: Group I contained undifferentiated stem cell grafts, group II contained adipocyte grafts, and group III contained osteoblast grafts. At weeks 4 and 8, in vivo fat formation was demonstrated in group II rats, as confirmed by Oil red O staining. At 8 weeks, group III rats demonstrated in vivo bone formation, as confirmed by the presence of osteocalcin on immunohistochemistry and the characteristic morphology of bone on hematoxylin-eosin staining. Group I rats demonstrated no in vivo bone or fat formation at either time interval. These results demonstrate the ability to isolate pluripotent stem cells from adipose tissue, to induce their differentiation into osteoblasts and adipocytes in vitro, and to form bone and fat subsequently in vivo. This is the first published report of in vivo bone formation from fat-derived stem cells. These cells may eventually serve as a readily available source of autologous stem cells for the engineering of bone and fat. PMID- 12783014 TI - Microarray gene analysis and expression profiles of Dupuytren's contracture. AB - Dupuytren's disease, although not altogether uncommon, has eluded scientists who have sought to explain the cause of this palmar fibroproliferative disorder. It can lead to severe limitations of hand function if left untreated. This study is the first broad genetic survey using microarray technology to find gene products that are overexpressed or underexpressed in diseased tissues. The authors found 23 genes with levels that differ consistently from control levels. Nine were selected for further verification using reverse transcription-polymerase chain reaction. These genes hold potential promise in explaining some of the demographic trends seen with disease, such as correlation with alcoholism and the striking predisposition for the male gender. PMID- 12783016 TI - Transfusion requirements for TRAM flap postmastectomy breast reconstruction. AB - The transverse rectus abdominis musculocutaneous (TRAM) flap is a commonly used method for autologous tissue postmastectomy breast reconstruction. It is a major operative procedure, and some have argued that it needlessly exposes patients to increased risk of complications and blood transfusions. In this series the authors review their experience with 105 consecutive complex postmastectomy breast reconstructions, limited to double-pedicle flaps, identifying complications rates and blood transfusion requirements. Blood transfusions were required in 2.8% of patients undergoing bipedicle or bilateral TRAM flap breast reconstruction. Blood transfusions were only required in patients who experienced a complication. Obesity was associated with an increased rate of complications and blood transfusion. Routine typing and cross-matching of blood and self donation of blood may not be required for TRAM flap breast reconstruction in low risk patients. PMID- 12783017 TI - Attitudes of adolescents about cosmetic surgery. AB - Although the number of teenagers choosing to have cosmetic procedures is increasing at a rapid rate, few studies have been published describing their attitudes about aesthetic surgery. To understand better their beliefs about cosmetic surgery, voluntary, confidential surveys were sent to the entire junior class of a suburban high school. Although two thirds of the respondents knew someone who had undergone cosmetic surgery, only one third would choose it for themselves. Those who desired aesthetic surgery described people who have cosmetic procedures as "motivated," whereas those who would not choose this option believed individuals who do so are "vain." The most desired procedures were liposuction, rhinoplasty, and breast augmentation. The main reasons for not proceeding were health risks, cost, and fear of a bad result. The most common source of information about plastic surgery among the students was teen magazines and television. PMID- 12783018 TI - Reconsidering the soleus muscle flap for coverage of wounds of the distal third of the leg. AB - Traditionally, cross-leg flaps and microsurgical flaps have been used to reconstruct defects of the distal third of the leg. In the authors' experience, the soleus muscle has also provided suitable tissue for coverage of these lesions in a notable number of cases. During a 2-year period, the authors treated 28 patients who required flap coverage of defects of the lower third of the leg. In this group, the soleus muscle was used successfully in 8 patients. All of these procedures resulted in healed wounds. The remaining patients underwent reconstruction with microsurgical flaps, fasciocutaneous local flaps, and a gastrocnemius muscle flap. Their experience has demonstrated that the soleus muscle is a valuable tool and should be included in the treatment algorithm for reconstructing lesions of the distal third of the lower extremity. PMID- 12783019 TI - Interdigitating fasciocutaneous gluteal V-Y advancement flaps for reconstruction of sacral defects. AB - The authors describe a modification of the classic gluteal bilateral V-Y advancement flap for sacral defect closure. After initial debridement, the V-Y design is marked on both sides of the defect. The incision is carried down to the fascia of the underlying gluteus maximus muscle. The upper and lower arms of the flaps are elevated and advanced on the gluteal muscle toward the midline, interdigitating each opposing arm. The overall result is a zigzag, broken midline suture. This procedure was carried out in 14 patients with sacral pressure sores and in 1 patient with a chronic pilonidal sinus. All flaps survived without major problems. There were no recurrences during the 6 to 16 months of follow-up. The interdigitating fasciocutaneous V-Y gluteal flap design is effective in breaking the midline vertical scar and preserving the gluteus maximus muscle. PMID- 12783020 TI - Seroma prevention using an albumin-glutaraldehyde-based tissue adhesive in the rat mastectomy model. AB - Seroma formation is a common postoperative complication in plastic surgery, resulting in notable patient morbidity and expense. In this study the authors investigated the efficacy of a synthetic glutaraldehyde-based tissue adhesive (BioGlue; Cryolife, Inc., Kennesaw, GA) in seroma prevention in the rat mastectomy seroma model. This tissue adhesive is composed of purified bovine serum albumin (45%) and glutaraldehyde (10%). Twenty-two Sprague-Dawley rats received mastectomies along with axillary lymphadenectomy and disruption of subcutaneous lymphatics. The control group (N = 11) received 2.5 ml saline into the wound before closure. In the experimental group (N = 11), 2.5 ml BioGlue was injected into the wound. On postoperative day 7, the resultant seromas were quantified and adjacent tissues were harvested for histological examination. The rats in the control group had a mean seroma volume of 5.19 ml (standard deviation, +/- 3.65 ml; N = 11), whereas the rats treated with BioGlue had a mean seroma volume of 0.25 +/- 0.43 ml (N = 11). Histology revealed an inflammatory response consistent with postoperative changes as well as a mild foreign body reaction. This study demonstrates that BioGlue can be used as a tissue adhesive that reduces seroma formation significantly in the rat mastectomy model (p = 0.0008). Further studies are warranted to investigate the long-term effect of BioGlue on surrounding tissue. PMID- 12783021 TI - Melkersson-Rosenthal syndrome in the periocular area: a review of the literature and case report. AB - A triad of facial palsy, facial edema, and furrowed tongue characterizes Melkersson-Rosenthal syndrome, a rare, noncaseating granulomatous disease of unknown cause. Although most reported cases of Melkersson-Rosenthal syndrome involve swelling of the perioral area, the authors present a case of Melkersson Rosenthal syndrome involving the periocular area. Because of its rarity, the syndrome is usually ignored and misdiagnosed; however, the syndrome should not only be considered in the classic perioral presentation but also in the rare periocular form, which may be confused with orbital tumors and orbital pseudotumors. Biopsies should be performed routinely in all patients who present with eyelid edema of unknown etiology. The physician and surgeon who see patients with head and neck pathology should be familiar with Melkersson-Rosenthal syndrome, and with the possibility of its presentation in the orbit and periocular region. PMID- 12783023 TI - Preoperative pneumoperitoneum used for tissue expansion before abdominal wall reconstruction. AB - The use of preoperative pneumoperitoneum was described initially for the repair of large ventral hernias. Large volumes of air are injected into the peritoneal cavity, which distends the abdominal wall progressively. This method is usually used to facilitate the reinstatement of the viscera into the abdominal cavity and improves postoperative diaphragmatic function. The authors use this method for preoperative tissue expansion of the abdominal wall before full-thickness resection of the abdominal wall for dermatofibrosarcoma protuberans in a 12-year old girl. In total, 6.8 L air was injected over 3 weeks without problem. The subsequent laxity of the abdominal wall permitted the primary closure of a 14- x 16-cm resection. The postoperative course was uncomplicated. Preoperative pneumoperitoneum can be used when reconstruction can be delayed. This is so in cases of slow tumor growth and in cases of infectious or posttraumatic full thickness abdominal defects. The parietal laxity obtained with the pneumoperitoneum not only increases the possibilities of direct closure but also increases the possibilities of pedicled flap reconstructions. PMID- 12783024 TI - Single-stage Achilles tendon reconstruction using a composite sensate free flap of dorsalis pedis and tendon strips of the extensor digitorum longus in a complex wound. AB - The reconstruction of the posterior heel including a wide defect of the Achilles tendon is difficult as a result of complicated infection, deficient soft tissue for coverage, and functional aspects and defects of the tendon itself. As a single-stage procedure, various methods of tendon transfer and tendon graft have been reported along with details of local flaps or island flaps for coverage. With advances in microsurgical techniques and subsequent refinements, several free composite flaps, including tendon, fascia, or nerve, have been used to reconstruct large defects in this area without further damaging the traumatized leg. The authors report such a single-stage reconstruction of a composite Achilles tendon defect using the extensor digitorum longus tendon of the second to fourth toe in combination with a dorsalis pedis flap innervated by the superficial peroneal nerve. The follow-up of this case has proved a satisfactory outcome to date. PMID- 12783025 TI - Desmoplastic melanoma of the nail. AB - Desmoplastic melanoma represents a variant of melanoma that is difficult to diagnose because 71% of patients have amelanotic skin lesions. In the acral region of the limbs, the clinical diagnosis is more difficult, especially in cases in which there are not clear, rapidly growing, pigmented nail streaks. Histopathological identification of desmoplastic melanoma is confusing because of the intense fibrous reaction in the dermis and minimal, atypical melanocytic proliferation at the dermal-epidermal junction. For these reasons, it is still misdiagnosed unfortunately as a variety of entities, including simple scar, fibrohistiocytic neoplasms, neural tumors, and superficial fibromatoses-with potentially devastating consequences. In equivocal cases, the use of immunohistochemistry (in particular S-100 and neuron-specific enolase) may be helpful in establishing the diagnosis. Because of the high local recurrence rate for desmoplastic melanoma of the finger, amputation is recommended in an effort to gain effective tumor control. Lymph node dissection may be reserved for patients with positive axillary nodes. PMID- 12783026 TI - Let's rock. PMID- 12783028 TI - Patient self-reported outcome after ulnar nerve transposition. PMID- 12783027 TI - Intrafocal pinning of distal radius fractures: a simplified approach. PMID- 12783029 TI - Circumcision as an unusual cause of penile lymphedema. PMID- 12783030 TI - About local anesthesia infiltration. PMID- 12783031 TI - Preemptive analgesia and a field block technique in reduction mammaplasty. PMID- 12783032 TI - Conchal graft. PMID- 12783034 TI - Ibuprofen and acetaminophen: effect on muscle inflammation after eccentric exercise. AB - PURPOSE: We examined the influence of ibuprofen and acetaminophen on muscle neutrophil and macrophage concentrations after novel eccentric contractions. METHODS: Twenty-four males (25 +/- 3 yr) were divided into three groups that received the maximal over-the-counter dose of either ibuprofen (1200 mg x d-1), acetaminophen (4000 mg x d-1), or a placebo after eccentric contractions of the knee extensors. Biopsies from the vastus lateralis were taken before and 24 h after exercise. Inflammatory cells were quantified in muscle cross-sections using immunohistochemistry. RESULTS: Macrophage concentrations were elevated by 1.5- to 2.5-fold (P < 0.05) at 24 h postexercise relative to preexercise concentrations, whereas neutrophil concentrations were not significantly elevated. Muscle inflammatory cell concentrations were unaffected by treatment with ibuprofen or acetaminophen when compared with placebo. CONCLUSIONS: Maximal over-the-counter doses of ibuprofen or acetaminophen, when administered therapeutically, do not affect muscle concentrations of neutrophils or macrophages 24 h after a novel bout of eccentric contractions. PMID- 12783035 TI - Gamma loop dysfunction in quadriceps on the contralateral side in patients with ruptured ACL. AB - PURPOSE: The aim of our study was to test for any neurophysiological abnormality in the gamma loop in the quadriceps femoris muscle on the uninjured side of patients with unilateral rupture of the anterior cruciate ligament (ACL). METHODS: Maximal voluntary contraction of knee extension and integrated electromyography (I-EMG) of the vastus medialis, vastus lateralis, and rectus femoris were measured in the uninjured limb of 13 patients with unilateral ACL rupture and 10 normal subjects, before and after 20-min vibration stimulation applied to the infrapatellar tendon. RESULTS: The mean percentage changes of maximal voluntary contraction and I-EMG in quadriceps femoris of the uninjured side of patients with ACL rupture were significantly different from those of the control group. Maximal voluntary contraction and I-EMG after prolonged vibration stimulation did not decrease as much as those of the control group even though the same protocol of vibration stimulation was applied. CONCLUSION: The abnormal response to prolonged vibration stimulation could represent abnormal gamma loop in the quadriceps femoris muscle of the uninjured side in patients with ACL rupture since the normal response of maximal voluntary contraction and I-EMG to prolonged vibration stimulation could not be evoked without normal function of the gamma loop. PMID- 12783036 TI - Vascular remodeling after spinal cord injury. AB - PURPOSE: Our purpose was to determine whether spinal cord injured (SCI) subjects have decreased femoral artery diameter and maximal hyperemic blood flow when expressed per unit of muscle volume compared with able-bodied (AB) individuals. A secondary purpose was to determine whether blood flow recovery rates were similar between groups. METHODS: Blood flow was measured in the femoral artery using Doppler ultrasound after distal thigh cuff occlusion of 4 and 10 min. Muscle mass of the lower leg was determined by magnetic resonance imaging (MRI). RESULTS: SCI individuals had smaller muscle cross-sectional areas (37%, P = 0.001) and volumes (38%, P = 0.001) than AB individuals. Furthermore, femoral artery diameter (0.76 +/- 0.14 vs 0.48 +/- 0.06 cm, AB vs SCI, P < 0.001) and femoral artery maximal blood flow (2050 +/- 520 vs 1220 +/- 240 mL x min-1, AB vs SCI, P < 0.001) were lower in SCI than AB individuals. Femoral artery diameter and maximal blood flow per unit muscle volume did not differ between SCI and AB individuals (P = 0.418 and P = 0.891, respectively). Blood flow recovery after ischemia was prolonged in SCI compared with AB individuals for both cuff durations (P = 0.048). CONCLUSIONS: In summary, femoral artery diameter and maximal hyperemic blood flow response per unit muscle volume are not different between SCI and AB individuals. Vascular atrophy after SCI appears to be closely linked to muscle atrophy. Furthermore, the SCI compared with AB individuals had a prolonged time to recovery, which may suggest decreased vessel reactivity. PMID- 12783037 TI - Subclassifying chronic fatigue syndrome through exercise testing. AB - PURPOSE: The purpose of this study was to examine physiological responses of persons with chronic fatigue syndrome (CFS) to a graded exercise test. METHODS: Cardiopulmonary exercise tests were performed on 189 patients diagnosed with CFS. Based on values for peak oxygen consumption, patients were assigned to one of four impairment categories (none, mild, moderate, and severe), using American Medical Association (AMA) guidelines. A one-way MANOVA was used to determine differences between impairment categories for the dependent variables of age, body mass index, percentage of predicted [OV0312]O(2), resting and peak heart rates, resting and peak systolic blood pressure, respiratory quotient (RQ), and rating of perceived exertion. RESULTS: Significant differences were found between each impairment level for percentage of predicted [OV0312]O(2) and peak heart rate. Peak systolic blood pressure values for the "moderate," and "severe" groups differed significantly from each other and both other groups. The more impaired groups had lower values. The no impairment group had a significantly higher peak RQ than each of the other impairment levels (all P < 0.001). Peak [OV0312]O(2) values were less than predicted for all groups. Compared with the males, the women achieved actual values for peak [OV0312]O(2) that were closer to their predicted values. CONCLUSION: Despite a common diagnosis, the functional capacity of CFS patients varies greatly. Stratifying patients by function allows for a more meaningful interpretation of the responses to exercise and may enable differential diagnosis between subsets of CFS patients. PMID- 12783038 TI - Head size and motor performance in children born prematurely. AB - PURPOSE: The objective of this study was to determine the relationship between head circumference (HC) and motor performance in 6- to 12-yr-old children born prematurely (PRE: birthweight < or = 1500 g, gestational age < or = 32 wk) and in children born at term (CON). METHODS: Thirty-three PRE and 21 CON without an apparent neurological deficit participated in this study. HC was measured on the day of testing and was rated as small HC (SHC, HC more than 1 SD below the 50th percentile of reference data) or as normal HC (NHC). Subjects were examined by an experienced neuropediatrician, and whole-body coordination was assessed by the Korper-Koordinationstest fur Kinder (KTK). Peak exercise performance was determined by a Wingate test and an incremental cycling test to volitional fatigue. Net oxygen cost of cycling was measured during four different tasks lasting 5-7 min each. Subjects pedaled at an intensity corresponding to 30% and 60% of peak oxygen uptake ([OV0312]O(2peak)) at a cadence of 36 and 76 rpm, respectively. RESULTS: Prematures with SHC showed no statistically significant difference in their neurological examination and whole-body coordination compared with prematures with NHC. Wingate test performance and [OV0312]O(2peak) relative to body mass were similar among SHC, NHC, and CON. In SHC, but not in NHC and CON, net oxygen cost of cycling increased significantly (P < 0.05) when cadence was increased from 36 to 76 rpm. CONCLUSION: At the age of 6-12 yr, SHC have a higher oxygen cost of cycling in exercise tasks requiring high velocity, which might be explained-at least in part-by an impaired neural control of intra- and intermuscular coordination. PMID- 12783039 TI - Effects of oral creatine and resistance training on myogenic regulatory factor expression. AB - PURPOSE: This study examined 12 wk of creatine (Cr) supplementation and heavy resistance training on skeletal muscle creatine kinase (M-CK) mRNA expression and the mRNA and protein expression of the myogenic regulatory factors Myo-D, myogenin, MFR-4, and Myf5. METHODS: Twenty-two untrained males were randomly assigned to either a control (CON), placebo (PLC), or Cr (CRT) group in a double blind fashion. Muscle biopsies were obtained before and after training. PLC and CRT trained thrice weekly using 3 sets of 6-8 repetitions at 85-90% 1-RM on the leg press, knee extension, and knee curl exercises. CRT ingested 6 g.d-1 of Cr for 12 wk while PLC consumed the equal amount of placebo. RESULTS: After training, M-CK mRNA expression, as well as myogenin and MRF-4 mRNA and protein expression, were found to be significantly greater for CRT compared with PLC and CON, whereas PLC was also significantly different from CON (P < 0.05). For Myo-D mRNA and protein, both CRT and PLC were significantly different from CON (P < 0.05), but CRT and PLC were not different from one another. No significant differences were located for Myf5 mRNA or protein (P > 0.05). M-CK mRNA was correlated with myogenin (r = 0.916) and MRF-4 (r = 0.883) protein (P < 0.05). CONCLUSION: When combined with heavy resistance training, Cr supplementation increases M-CK mRNA expression, likely due to concomitant increases in the expression of myogenin and MRF-4. Therefore, increases in myogenin and MRF-4 mRNA and protein may play a role in increasing myosin heavy chain expression, already shown to occur with Cr supplementation. PMID- 12783040 TI - Plasma creatine kinase activity and glutathione after eccentric exercise. AB - PURPOSE: This study examined whether plasma total glutathione levels could explain the intersubject variability in the creatine kinase (CK) response to eccentric exercise. We hypothesized that the increase in plasma CK activity after eccentric exercise would be lower for individuals with low plasma total glutathione (<2.5 micromol x L-1) compared with individuals with high total glutathione (>3.8 micromol x L-1), but other indicators of muscle damage would be the same between groups. METHODS: Resting blood samples were obtained over 2 d from 60 subjects and analyzed for plasma total glutathione. Eight subjects who had total glutathione values below 2.5 micromol x L-1 (LG), and nine who had values above 3.8 micromol x L-1 (HG) performed 50 maximal eccentric actions of the elbow flexors. Maximal voluntary isometric contraction (MVC), relaxed arm angle (RANG), and blood samples for CK, myoglobin (Mb), and total glutathione were obtained pre, post (except blood samples), 24, 48, 72, 96, and 120 h after exercise. RESULTS: There was a significant group-by-time interaction in analysis of MVC, RANG, total glutathione, CK, and Mb response to exercise. Although LG showed a smaller CK response to eccentric exercise compared with HG, LG also showed a smaller increase in plasma Mb, a faster recovery of MVC and RANG, and an increase in plasma total glutathione. CONCLUSION: Subjects with low plasma total glutathione levels had a smaller plasma CK and Mb response and a faster recovery from eccentric exercise compared with subjects having high plasma total glutathione levels. We suggest that a blunted inflammatory response in subjects with low plasma glutathione may be one explanation for these findings. PMID- 12783041 TI - Hormonal responses from concentric and eccentric muscle contractions. AB - Intense resistance exercise can acutely increase testosterone (T), free testosterone (FT), and growth hormone (GH) concentrations, but there are few investigations concerning acute endocrine responses to concentric (CON) and eccentric (ECC) contractile actions. PURPOSE: The purpose of the study was to compare acute anabolic hormonal responses to bouts of dynamic CON and ECC contractions from multiple exercises at the same absolute load. METHODS: Ten young men (age: 24.7 +/- 1.2 yr, weight: 85.45 +/- 24.2 kg, and height: 178 +/- 0.2 cm) completed two trials in counterbalanced fashion consisting of only CON or ECC contractions at the same absolute workload. Subjects performed four sets of 12 repetitions of bench press, leg extension, military press, and leg curl at 80% of a 10-repetition maximum with 90-s rest periods. Blood samples were collected pre-, post-, and 15-min postexercise. RESULTS: There were significant increases in GH, T, and FT and lactate for both trials, but only GH and lactate were greater for the CON trial. CONCLUSION: CON exercise increases GH concentrations to a much greater extent than ECC exercise at the same absolute load, and it is likely that greater GH responses were related to intensity rather than mode of contraction. Also, CON and ECC dynamic contraction trials at the same absolute workload elicited similar small but significant increases in T and FT, indicating that the greater metabolic stress produced by during the CON trial did not affect these hormone responses. PMID- 12783042 TI - Functional adaptability of muscle fibers to long-term resistance exercise. AB - PURPOSE: We compared the functional properties of muscle fibers from two groups of subjects that differed widely in their training history to investigate whether long-term resistance exercise alters the intrinsic contractile properties of skeletal muscle fibers. METHODS: Vastus lateralis muscle biopsies were obtained from six sedentary males (NT group, age = 23 +/- 1 yr) and six males who had participated in regular resistance exercise training over the preceding 7.6 +/- 1.6 yr (RT group, 22 +/- 1 yr). Chemically skinned muscle fiber segments were activated with a saturating free [Ca2+] to quantify fiber peak Ca2+-activated force (P(o)), unloaded shortening velocity (V(o)), and peak power. Fiber segment myosin heavy chain (MHC) isoform content was identified by gel electrophoresis. RESULTS: Slow and fast fibers from the RT group were larger in CSA and produced greater absolute P(o) and absolute peak power in comparison with fibers from the NT group. However, these differences were no longer evident after P(o) and peak power were normalized to fiber CSA and fiber volume, respectively. V(o)/fiber length was dependent on fiber MHC content but independent of training status. CONCLUSION: Fiber hypertrophy was sufficient to account for intergroup differences in P(o) and peak power of slow and fast fibers. There was no evidence that the intrinsic contractility of slow or fast fibers, as evaluated by force, shortening velocity, and power normalized to the appropriate fiber dimensions, differed between RT and NT groups. PMID- 12783043 TI - Effects of dietary fat on muscle substrates, metabolism, and performance in athletes. AB - INTRODUCTION: The present investigation aimed at identifying differences in muscle structural composition, substrate selection, and performance capacity in highly trained endurance athletes as a consequence of consuming a high-fat or a low-fat diet. METHODS: Eleven duathletes ingested high-fat (53% fat; HF) or high carbohydrate diets (17% fat; LF) for 5 wk in a randomized crossover design. RESULTS: In m. vastus lateralis, oxidative capacity estimated as volume of mitochondria per volume of muscle fiber (HF: 9.86 +/- 0.36 vs LF: 9.79 +/- 0.52%, mean +/- SE) was not different after the two diet periods. Intramyocellular lipid (IMCL) was significantly increased after HF compared with LF (1.54 +/- 0.27% vs 0.69 +/- 0.09%, P = 0.0076). Glycogen content was lower after HF than after LF, but this difference was not statistically significant (487.8 +/- 38.2 vs 534.4 +/ 32.6 mmol x kg-1 dry weight, P = 0.2454). Maximal power and [OV0312]O(2max) (63.6 +/- 0.9 vs 63.9 +/- 1.2 mL O(2) x min-1 x kg-1 on HF and LF) during an incremental exercise test to exhaustion were not different between the two diet periods. Total work output during a 20-min all-out time trial (298 +/- 6 vs 297 +/- 7 W) on a bicycle ergometer as well as half-marathon running time (80 min 12 s +/- 86 s vs 80 min 24 s +/- 82 s) were not different between HF and LF. Blood lactate concentrations and respiratory exchange ratios (RER) were significantly lower after HF than after LF at rest and during all submaximal exercise loads. CONCLUSIONS: Muscle glycogen stores were maintained after a 5-wk high-fat diet period whereas IMCL content was more than doubled. Endurance performance capacity was maintained at moderate to high-exercise intensities with a significantly larger contribution of lipids to total energy turnover. PMID- 12783044 TI - Influence of short-term endurance exercise training on heart rate variability. AB - PURPOSE: To examine the influence of 2 wk (eight sessions) of endurance training on cardiac autonomic modulation, as measured by heart rate variability (HRV). METHODS: Twenty-four males (mean age: 23.1 yr) were randomized to an exercise (EX; N = 12) or control group (CT; N = 12). EX trained for eight sessions (4x wk 1, 40 min, 80-85% HRreserve) on a cycle ergometer. ECG tracings were collected during 5 min of paced breathing (12 breaths x min-1 (PB)), 5 min of spontaneous breathing (SB1), 5 min of 70 degrees head-up tilt (TILT), and a second 5-min period of spontaneous breathing (SB2). Data were collected before (test 1), during (tests 2-4), and 48 h after (test 5) the 2-wk period. HRV was reported as the standard deviation of RR intervals, and as natural logarithm of the normalized units (NU) of high- and low-frequency power (lnHF and lnLF). RESULTS: EX exhibited a significant increase in peak oxygen consumption (8%). During PB and TILT conditions, ANOVA revealed a group x time interaction such that EX exhibited lower lnLFNU and lnLF/lnHF during test 5 compared with test 1. CONCLUSION: These data suggest that eight endurance exercise-training sessions performed over 2 wk enhance the relative vagal modulation of the heart during PB and TILT, but not during SB. PMID- 12783045 TI - Baroreflex-mediated heart rate and vascular resistance responses 24 h after maximal exercise. AB - INTRODUCTION: Plasma volume, heart rate (HR) variability, and stimulus-response relationships for baroreflex control of forearm vascular resistance (FVR) and HR were studied in eight healthy men after and without performing a bout of maximal exercise to test the hypotheses that acute expansion of plasma volume is associated with 1) reduction in baroreflex-mediated HR response, and 2) altered operational range for central venous pressure (CVP). METHODS: The relationship between stimulus (DeltaCVP) and vasoconstrictive reflex response (DeltaFVR) during unloading of cardiopulmonary baroreceptors was assessed with lower-body negative pressure (LBNP, 0, -5, -10, -15, -20 mm Hg). The relationship between stimulus (Deltamean arterial pressure (MAP)) and cardiac reflex response (DeltaHR) during loading of arterial baroreceptors was assessed with steady-state infusion of phenylephrine (PE) designed to increase MAP by 15 mm Hg alone and during application of LBNP (PE+LBNP) and neck pressure (PE+LBNP+NP). Measurements of vascular volume and autonomic baroreflex responses were conducted on two different test days, each separated by at least 1 wk. On one day, baroreflex response was tested 24 h after graded cycle exercise to volitional exhaustion. On another day, measurement of baroreflex response was repeated with no exercise (control). The order of exercise and control treatments was counterbalanced. RESULTS: Baseline CVP was elevated (P = 0.04) from a control value of 10.5 +/- 0.4 to 12.3 +/- 0.4 mm Hg 24 h after exercise. Average DeltaFVR/DeltaCVP during LBNP was not different (P = 0.942) between the exercise (-1.35 +/- 0.32 pru x mm Hg-1) and control (-1.32 +/- 0.36 pru x mm Hg-1) conditions. However, maximal exercise caused a shift along the reflex response relationship to a higher CVP and lower FVR. HR baroreflex response (DeltaHR/DeltaMAP) to PE+LBNP+NP was lower (P = 0.015) after maximal exercise (-0.43 +/- 0.15 beats x min-1 x mm Hg-1) compared with the control condition (-0.83 +/- 0.14 beats x min-1 x mm Hg-1). CONCLUSION: Expansion of vascular volume after acute exercise is associated with altered operational range for CVP and reduced HR response to arterial baroreceptor stimulation. PMID- 12783046 TI - Sympathoadrenergic mechanisms in reduced hemodynamic stress responses after exercise. AB - PURPOSE: This study examines the acute effects of moderate aerobic exercise on 1) hemodynamic and sympathetic activity during behavioral stress and 2) beta adrenergic receptor responsivity in a biracial sample of 24 sedentary adults. METHODS: Before and after exercise, blood pressure (BP), impedance-derived cardiovascular measures, and plasma norepinephrine (NE) and epinephrine (EPI) were assessed during mental arithmetic and active speech tasks, and beta adrenergic receptor responsivity was assessed using a standard isoproterenol challenge procedure. RESULTS: After exercise, BP, NE, and EPI responses to stress were reduced (0.0001 < P < 0.08), preejection period (PEP) was elongated (P < 0.0001), and beta(1)- and beta(2)-receptor responsivity (P < 0.02) was enhanced. Approximately 65% of the prepost exercise mean arterial pressure response difference could be accounted for by changes in sympathetic factors, with change in NE and PEP being the single best predictors. CONCLUSIONS: Reduced BP responses to stress after acute exercise are strongly linked to a decrease in sympathetic drive, as evidenced by reduced NE responses and elongation of the PEP. Coincident with this overall dampening of the hemodynamic response to stress, increases in cardiac and vascular beta-adrenergic receptor responsivity occur. These findings may have important implications for future translational studies that seek to articulate the mechanisms through which regular aerobic exercise reduces the risks of hypertensive and coronary heart disease. PMID- 12783047 TI - Effects of ephedrine, caffeine, and their combination on muscular endurance. AB - PURPOSE: The purpose of this study was to investigate the effects of ingesting caffeine (C), ephedrine (E), and their combination on muscular endurance, using a double-blind, repeated measures design. METHODS: Ninety minutes after ingesting either C (4 mg x kg-1), E (0.8 mg x kg-1), a combination of C+E, or a placebo (P), 13 male subjects performed a weight-training circuit consisting of three supersets (SS), each SS consisting of leg press (at 80% of 1 RM to exhaustion) followed by bench press (at 70% 1-RM to exhaustion); 2 min of rest intervened between SS. RESULTS: The trials involving ephedrine ingestion (C+E and E), when compared with the nonephedrine trials (C and P), caused significant increases (P < 0.05) in the mean number of repetitions completed for both the leg-press and bench-press exercises but only during the first SS. During that first set, the mean number (+/-SD) of repetitions for leg press was 19 +/- 8, 16 +/- 7, 14 +/- 6, and 13 +/- 5 for the C+E, E, C, and P trials, respectively. The mean numbers of repetitions for the first set of bench-press exercise were 14 +/- 3, 13 +/- 3, 12 +/- 3, and 12 +/- 3 for the C+E, E, C, and P trials, respectively. As a result, the total weight lifted during all three sets was greater for the trials involving ephedrine ingestion. Systolic blood pressure before exercise was significantly increased with both ephedrine treatment trials when compared with the other trials (C+E = 156 +/- 29 mm Hg; E = 150 +/- 14; C = 141 +/- 16; P = 138 +/- 14). CONCLUSION: It was concluded that acute ingestion of C+E and E increases muscular endurance during the first set of traditional resistance-training exercise. The performance enhancement was attributed primarily to the effects of E; there was no additive effect of C. PMID- 12783048 TI - Impact of high-intensity exercise on nitric oxide exchange in healthy adults. AB - PURPOSE: After exercise, exhaled NO concentration has been reported to decrease, remain unchanged, or increase. A more mechanistic understanding of NO exchange dynamics after exercise is needed to understand the relationship between exercise and NO exchange. METHODS: We measured several flow-independent NO exchange parameters characteristic of airway and alveolar regions using a single breath maneuver and a two-compartment model (maximum flux of NO from the airways, J'(awNO), pL x s-1; diffusing capacity of NO in the airways, D(awNO), pL x s-1 x ppb-1; steady state alveolar concentration, C(alv,ss), ppb; mean airway tissue NO concentration, C(awNO), ppb), as well as serum IL-6 at baseline, 3, 30, and 120 min after a high-intensity exercise challenge in 10 healthy adults (21-37 yr old). RESULTS: D(awNO) (mean +/- SD) increased (37.1 +/- 44.4%), whereas J'(awNO) and C(awNO) decreased (-7.27 +/- 11.1%, -26.1 +/- 24.6%, respectively) 3 min postexercise. IL-6 increased steadily after exercise to 481% +/- 562% above baseline 120 min postexercise. CONCLUSION: High-intensity exercise acutely enhances the ability of NO to diffuse between the airway tissue and the gas phase, and exhaled NO might be used to probe both the metabolic and physical properties of the airways. PMID- 12783049 TI - Seasonal variation in adult leisure-time physical activity. AB - PURPOSE: To determine the effect of season on self-reported leisure-time physical activity (LTPA) behaviors of Michigan adults. METHODS: Data were obtained from the 1996 Michigan Behavioral Risk Factor Survey conducted throughout the year. Survey respondents were considered active if they reported participating in at least one LTPA during the past month. Complete information regarding type, frequency, and duration of up to two LTPA was available on 2843 adults (1635 women and 1208 men). Four seasons were defined as winter (January-March; N = 677), spring (April-June; N = 759), summer (July-September; N = 760), and fall (October-December; N = 647). Total weekly leisure-time energy expenditure was quantified (kcal x kg-1 x wk-1) from MET intensities, duration, and frequency of activity sessions per week. Seasonal differences were identified using ANOVA. RESULTS: Average (+/-SEM) weekly leisure time energy expenditure was significantly greater (P < 0.001) during spring (17.5 +/- 0.8 kcal x kg-1 x wk-1) and summer (17.5 +/- 0.7 kcal.kg-1.wk-1) compared with winter (14.8 +/- 0.7 kcal x kg-1 x wk-1) and fall (15.0 +/- 0.7 kcal x kg-1 x wk-1). Duration of the first activity was significantly greater (P < 0.05) in summer (58.6 +/- 1.6 min) compared with winter (53.4 +/- 1.8 min). However, intensity (4.6 +/- 0.1 METs) and frequency (3.1 +/- 0.1 sessions per week) of the first activity did not differ among seasons. A second activity was performed by 1319 (46.4%) of active individuals and was more common in the spring (46.8%) and summer (54.5%) compared with fall (42.6%) and winter (39.4%) (chi2 = 31.0; P < 0.01). When both active and inactive subjects are considered, the Healthy People 2010 recommendation for moderate physical activity was met only during spring and summer. CONCLUSION: Weekly leisure-time energy expenditure averaged approximately 15-20% higher during spring and summer. Much of this difference was due to active respondents participating in a second activity during these seasons. PMID- 12783050 TI - Symmetry and reproducibility of kinematic parameters during various running techniques. AB - PURPOSE: This study examined the validity of the assumption of lower-extremity kinematic parameter reproducibility and symmetry during running with different velocities and stride frequencies. METHODS: Each of 12 female long-distance runners ran on a treadmill in combinations of three different velocities (2.5, 3.0, and 3.5 m.s-1) and three different stride frequencies (preferred and +/- 10% from preferred). The left and right sides of the athletes were filmed using video cameras placed orthogonally to the sagittal plane. A total number of three step cycles for each running condition were recorded (250 Hz). For each side of the body, 19 sagittal kinematic parameters from the lower extremity were evaluated. RESULTS: Intraclass correlation coefficients (ICC) for both legs were high (generally > 0.80). Only the angular velocity parameters demonstrated correlation values below 0.70. The symmetry index for the linear and angular displacement parameters and the contact times during all running techniques were less than 8%, whereas those for the angular velocity parameters and flight times were higher than 15%. CONCLUSION: The present results suggested that the degree of reproducibility and symmetry of kinematic data do not vary with a deliberate change in running technique but rather depend on the parameter itself. With respect to the economy of data analysis, the present findings indicate that recording a single monolateral trial would provide reproducible and symmetric values for most kinematic parameters. PMID- 12783051 TI - RPE, pain, and physiological adjustment to concentric and eccentric contractions. AB - PURPOSE: The purpose of the study was to compare perceptual (RPE and pain), cardiac (heart rate), lactate, and endocrine (cortisol) responses with concentric (CON) and eccentric (ECC) resistance exercise protocols using the same absolute workload. METHODS: Eight healthy men with resistance-training experience participated in the study. Subjects completed two experimental trials consisting of either CON contractions or ECC contractions at the same absolute workload for each of four exercises: bench press, leg extension, military press, and leg curl. Subjects performed four sets of 12 repetitions at 80% of 10-RM with 90-s rest periods. Blood samples were taken before, immediately after, and 15-min postexercise. RESULTS: There was a significant trial effect for RPE, with CON exercise eliciting a higher RPE than ECC exercise (6.71 +/- 0.51 and 4.10 +/- 0.27, respectively). A significant trial effect was also demonstrated for pain, with CON exercise producing a higher pain rating than ECC exercise (5.59 +/- 0.41 and 3.23 +/- 0.27, respectively). Significantly higher heart rates and lactates were also demonstrated during the CON trial. For cortisol, a significant interaction was revealed between the pre- and immediate posttrial measures but not an overall trial effect. Correlational analyses revealed a significant relationship between RPE and pain for both trials. CONCLUSIONS: CON exercise elicits greater perceptual (higher RPE and pain rating), cardiac, lactate and cortisol response than ECC exercise at the same absolute workload. Data demonstrate that relative to absolute load, RPE and pain respond to resistance exercise in a similar fashion. Additionally, physiological cues are consistent with these perceptual data. PMID- 12783052 TI - The effects of graded resistance exercise on platelet aggregation and activation. AB - PURPOSE: The purpose of this study was to examine the effects of resistance exercise with varying intensity but with similar volume on platelet aggregation and activation. METHODS: Thirteen healthy male subjects randomly completed three resistance exercise test trials at an intensity corresponding to 40%, 60%, and 80% of one repetition maximum (1-RM) in which the subjects performed six exercises including upper- and lower-body parts. Venous blood samples were obtained before and immediately after each exercise trial and analyzed for platelet count (PLT), plateletcrit (PCT), mean platelet volume (MPV), platelet aggregation, and beta-thromboglobulin (B-TG). Plasma volume changes were estimated from hemoglobin and hematocrit readings before and after each exercise trial. RESULTS: Although all exercise trials were followed by a significant (P < 0.05) increase in PLT (thrombocytosis), PCT, and MPV, this rise was not related to the exercise intensity (P > 0.05). Exercise was also followed by a significant increase (P < 0.05) in platelet aggregation, but this only occurred with the high but not with the low concentrations of adenosine diphosphate (ADP). Although ANOVA showed a significant overall increase (P < 0.05) in the concentration of B TG after exercise, this rise only reached the assigned level of significance (P < 0.05) after 80% exercise trial. CONCLUSION: It was concluded therefore that resistance exercise is followed by an increase in PLT, PCT, and MPV, and this occurred in parallel with an in vivo activation of platelet as manifested by an increase in platelet aggregation and a rise in B-TG. PMID- 12783054 TI - Neuromuscular response of young boys versus men during sustained maximal contraction. AB - PURPOSE: This study was designed to compare neuromuscular response between boys and men during sustained maximal voluntary contraction (MVC). METHODS: Fifteen boys (YB, 10.5 +/- 0.9 yr) and 12 men (AM, 21.5 +/- 4.5 yr) participated in the experiment. Arm's cross sectional area (CSA) and maximal force (F(max)) of elbow flexor were measured before subjects performed a 30-s sustained MVC. Mean power frequency (MPF) and muscle fiber conduction velocity (MFCV) were calculated from myoelectric signals of the biceps brachii. F(max)/CSA, MPF, and MFCV changes were expressed by slopes of linear regressions. Maximal MPF (I-MPF) and MFCV (I-MFCV) were derived from the intercept of each regression. RESULTS: AM had significantly greater F(max)/CSA (P < 0.05), I-MPF (P < 0.05), and I-MFCV (P < 0.01) than YB. F(max)/CSA (P < 0.001), MPF (P < 0.001), and MFCV (P < 0.01) declined significantly more for AM than YB. MPF/MFCV ratio increased, i.e., MPF decreased more than MFCV, for both groups but this was significantly (P < 0.001) more pronounced for AM. CONCLUSION: Taken together, those results suggest that more fatigable Type II motor units are involved in men, resulting in greater lactic acid and ions accumulations during fatigue. This difference in muscle's metabolic and ionic state could be responsible for a greater reflex-induced decrease of motor units firing rates in men compared with boys. This firing rate decrease could be explained using the "muscular wisdom" hypothesis and would express a nervous command adaptation to sustain a maximal contraction. PMID- 12783053 TI - Strength increase after whole-body vibration compared with resistance training. AB - PURPOSE: The aim of this study was to investigate and to compare the effect of a 12-wk period of whole-body vibration training and resistance training on human knee-extensor strength. METHODS: Sixty-seven untrained females (21.4 +/- 1.8 yr) participated in the study. The whole-body vibration group (WBV, N = 18) and the placebo group (PL, N = 19) performed static and dynamic knee-extensor exercises on a vibration platform. The acceleration of the vibration platform was between 2.28 g and 5.09 g, whereas only 0.4 g for the PL condition. Vibration (35-40 Hz) resulted in increased EMG activity, but the EMG signal remained unchanged in the PL condition. The resistance-training group (RES, N = 18) trained knee extensors by dynamic leg-press and leg-extension exercises (10-20 RM). All training groups exercised 3x wk-1. The control group (CO, N = 12) did not participate in any training. Pre- and postisometric, dynamic, and ballistic knee-extensor strength were measured by means of a motor-driven dynamometer. Explosive strength was determined by means of a counter-movement jump. RESULTS: Isometric and dynamic knee-extensor strength increased significantly (P < 0.001) in both the WBV group (16.6 +/- 10.8%; 9.0 +/- 3.2%) and the RES group (14.4 +/- 5.3%; 7.0 +/- 6.2%), respectively, whereas the PL and CO group showed no significant (P > 0.05) increase. Counter-movement jump height enhanced significantly (P < 0.001) in the WBV group (7.6 +/- 4.3%) only. There was no effect of any of the interventions on maximal speed of movement, as measured by means of ballistic tests. CONCLUSIONS: WBV, and the reflexive muscle contraction it provokes, has the potential to induce strength gain in knee extensors of previously untrained females to the same extent as resistance training at moderate intensity. It was clearly shown that strength increases after WBV training are not attributable to a placebo effect. PMID- 12783055 TI - The size-independent oxygen cost of running. AB - PURPOSE: The purpose of this investigation was to determine whether differences in running economy among children, adolescents, and adults can be explained by differences in resting metabolism, mass, and stature. METHODS: Participants were 36 children, 23 adolescents, and 24 adults. Mass-specific gross oxygen cost per minute ([OV0312]O(2gross) x M-1), mass-specific gross oxygen cost per kilometer (VO(2gross) x M-1), mass-specific net oxygen cost per kilometer (VO(2net) x M-1), and a dimensionless index called the size-independent cost (SIC) were compared for level treadmill running at speeds ranging from 1.6 to 3.1 m.s-1. SIC was defined as the net oxygen cost to move a mass of 1 kg a distance equal to stature (mL x kg-1). RESULTS: Children generally had higher [OV0312]O(2gross).M-1, VO(2gross) x M-1, and VO(2net) x M-1 than adolescents who similarly had greater costs than adults. When SIC was used to control for size-related differences in resting metabolism, mass and stature the costs of children and adults were similar (0.323 +/- 0.034 and 0.338 +/- 0.035 mL x kg-1, respectively, P = 0.54). However, adolescents had significantly higher SIC (0.360 +/- 0.026 mL x kg-1, P < 0.001) than both children and adults. Analysis of data from the literature indicated SIC peaks around 15 yr of age and changes were parallel to changes in the ratio of leg length to stature. CONCLUSIONS: We conclude that when resting metabolism and the dimensional effects of mass and stature are controlled, the running economy of adolescents is greater than in children and adults, which are similar. Therefore, differences in [OV0312]O(2gross) x M-1, VO(2gross) x M-1, and VO(2net) x M-1 among children, adolescents, and adults do not solely reflect qualitative differences in running performance. PMID- 12783056 TI - Laboratory calibration and validation of the Biotrainer and Actitrac activity monitors. AB - PURPOSE: The Biotrainer and Actitrac activity monitors (IM Systems) offer potential research advantages over existing accelerometry-based activity monitors, but they have not been tested under controlled conditions. The purpose of this study was to develop and test laboratory-based prediction equations for both monitors to estimate energy expenditure (EE) for walking/running movements. METHODS: Participants in the study wore a Biotrainer and Actitrac monitor on both hips and completed three paced bouts on the treadmill (3, 4, and 6 mph for 6 min each). Metabolic data collected using an indirect calorimetry system were used as the criterion measure. Multiple regression techniques were performed to develop prediction equations, and these equations were then applied to data from a separate sample for cross-validation purposes. Reliability was also examined. RESULTS: The correlations between the raw counts from each monitor and the measured metabolic variables ranged from r = 0.74-0.88 for the Biotrainer and from r = 0.81-0.91 for the Actitrac. The equations predicting EE (kcal x min-1) from counts yielded strong validation results for both the Biotrainer (R2 = 0.88, SEE = 1.47) and the Actitrac (R2 = 0.91, SEE = 1.24). When used on the cross validation sample, the correlations between measured and predicted EE were r = 0.93 (Biotrainer) and r = 0.94 (Actitrac). Intraclass reliability coefficients computed between the left and right monitors ranged from 0.60 to 0.71 (Biotrainer) and 0.80 to 0.87 (Actitrac). When the equation developed from one side was applied to data from the monitor on the other side, there were no significant differences in predicted and measured EE for most comparisons. CONCLUSION: The results support the validity of Biotrainer and Actitrac monitors for estimating energy expenditure under controlled conditions. PMID- 12783057 TI - Quantifying physical activity via pedometry in elementary physical education. AB - PURPOSE: The objective of this study was to determine a pedometer steps per minute standard for quantifying the lesson time that first- and second-grade physical education students spent in moderate to vigorous physical activity (MVPA). METHODS: The sample was divided into validation (N = 246) and cross validation (N = 123) samples using the holdout technique. Using the criterion test model, steps per minute cut points were empirically and judgmentally determined. C-SOFIT systematic observation was the criterion instrument and pedometry was the predictor instrument. Data were collected from 45 physical education lessons implemented in six schools. The three-step analytic procedure of computing mastery/nonmastery outcome probabilities, phi coefficients, and error proportions was used to determine the optimal steps per minute cut point for quantifying 33.33% of the physical education lesson time engaged in MVPA within a 30 class. RESULTS: Steps per minute was highly correlated with observation (r = 0.74-0.86, P < 0.0001). Five steps per minute scores that were accurate indicators of 33.33% of the class time engaged in MVPA in the validation sample were accurate indicators of steps per minute in the cross-validation sample. The optimal steps per minute cut point ranged from 60.00 to 63.00, which is equivalent to 1800-1890 steps in a 30-min physical education class. CONCLUSION: Data supports the use of pedometry steps per minute values as an accurate indicator of MVPA. Pedometry demonstrates promise as a viable large scale surveillance instrument for measuring MVPA in physical education. PMID- 12783058 TI - Exercise and oxidation of dietary fat. PMID- 12783060 TI - Response to a plaque control regimen on different levels of gingival inflammation. AB - BACKGROUND: The purpose of the present study was to evaluate the effectiveness of an oral hygiene regimen in subjects presenting with substantially different severity of plaque-associated gingivitis. METHODS: The study population was selected from among a large pool of subjects undergoing an experimental gingivitis trial. At completion of the 21-day plaque accumulation period, 2 sub groups of subjects were identified on the basis of uppermost and lowest quartile for Gingival Index (GI), respectively classified as highly-inflamed (Hinf; n=17; GI: 1.07+/-0.10) and slightly-inflamed (Sinf; n=22; GI: 0.28+/-0.09) groups. An oral hygiene regimen, based on use of amine fluoride/stannous fluoride-containing toothpaste and mouthrinse, was then prescribed for 21 days. RESULTS: Plaque Index (PI), GI, gingival crevicular fluid (GCF) and Angulated Bleeding Index (AngBI) significantly decreased after treatment in both HInf and SInf groups (p<0.001). However, PI (0.77+/-0.41 vs 0.43+/-0.33, p<0.01), GI (0.23+/-0.30 vs 0.08+/-0.11, p<0.05), GCF (15.23 +/-7.11 vs 7.66+/-2.93, p<0.0000) remained significantly greater in the Hinf group compared to the Sinf group. CONCLUSIONS: These results suggest that 1) an oral hygiene regimen based on amine/stannous fluoride containing toothpaste and mouthrinse is effective in reducing plaque-associated gingivitis, regardless of pre-existing severity of gingival inflammation; 2) the level of improvement in gingival status, however, is dependent on the pre existing severity of the inflammatory condition. PMID- 12783061 TI - Immunophenotypic analysis of human gingival fibroblasts and its regulation by Granulocyte-Macrophage Colony-Stimulating Factor (GM-CSF). AB - BACKGROUND: The aim of the present study was to perform an immunophenotypic analysis of human gingival fibroblast cells and its eventual modulation by Granulocyte-Macrophage Colony-Stimulating Factor (GM-CSF). METHODS: Gingival fibroblasts were derived from gingival biopsy of 15 healthy subjects. The presence of fibroblast cells in culture and the absence of epithelial cells was performed with fluorescence microscopy using vimentin and cytokeratin markers, respectively. Molecular expression of gingival fibroblast cell membrane was carried out with monoclonal antibodies by flow cytometry analysis. Human recombinant GM-CSF at the concentration of 200 ng/ml was used for the in vitro stimulation of gingival fibroblasts. Statistical analysis was performed using the Student "t"-test. RESULTS: Human gingival fibroblasts express a wide surface molecular panel including mainly CD59, CD99, CD9, CD95, CD55, CD63, CD26, CD117, CD71 and CD86. The GM-CSF seems to regulate the CD49B expression positively and the CD40 and CD103 expression negatively. CONCLUSIONS: Results show that GM-CSF is able to modulate the in vitro expression of some membrane molecules of gingival fibroblasts and therefore it may regulate, in vivo, peculiar specific biological functions of gingival tissue. PMID- 12783062 TI - The role of ultrasonography in the diagnosis of temporomandibular joint disc displacement and intra-articular effusion. AB - AIM: The aim of this work is to conduct a preliminary study to investigate the usefulness of ultrasonography (US) in the study of temporomandibular joint (TMJ), comparing ultrasonographic diagnosis of joint effusion and disc displacement with those based on an accurate clinical examination. METHODS: Participants in this study were 47 consecutive patients complaining for TMJ problems. All 94 TMJs were evaluated to detect the presence of intra-articular effusion and disc displacement by means of 2 diagnostic instruments: a standardized clinical assessment based on the Research Diagnostic Criteria for Temporomandibular Disorders (RDC-TMD) at the Section of Prosthetic Dentistry, and an ultrasonographic investigation conducted by a blinded operator at the Operative Unit of the Department of Rheumatology, Univesity of Pisa, Italy. Agreement between the two diagnostic techniques has been evaluated by means of Cohen's K test. RESULTS: Ultrasonography showed a good agreement with clinical assessment for the diagnosis of both intra-articular effusion (percentage of agreement 80%; K=0.611) and disc displacement (agreement 81.9%; K=0.572). CONCLUSION: When compared to a standardized clinical assessment, ultrasonographic technique showed a good diagnostic capability to detect TMJ intra-articular effusion and disc displacement. Within all the limitations of this study, it can be suggested that ultrasonography could represent a promising imaging technique in the study of temporomandibular joint. PMID- 12783063 TI - [Clinical comparison between guided tissue regeneration and induced tissue regeneration]. AB - AIM: The aim of the present study was to evaluate, over a period of 1 y, 3 different surgical methods for the treatment of periodontal bone defects. METHODS: Thirty-six infrabone defects, at least 4 mm in depth, in non-smokers were enrolled in the study. Of these, 12 were treated with guided tissue regeneration (GTR) using a resorbable membrane with collagen, 12 with enamel matrix derivative (EMD) gel and 12 with a modified Widman flap (MWF). The pocket depth, attachment loss and gingival recession both before commencement of therapy and after an interval of 1 y are reported. RESULTS: The reduction in pocket depth was 4 mm, 4.4 mm, and 4.5 mm, respectively, for the control group, GTR group and EMD group. The attachment gain for the respective treatments was: MWF, 2 mm; GTR, 2.8 mm; EMD, 2.9 mm. Gingival recession was: MWF, 1.8 mm; GTR, 1.5 mm; EMD, 1.1 mm. CONCLUSION: These findings show the efficacy of the 3 methods in the treatment of bone defects, but none of the 3 emerges as being statistically superior to the others 2. PMID- 12783064 TI - [Corrosion of titanium in presence of dental amalgams and fluorides]. AB - AIM: The aim of this study was to evaluate the behaviour of titanium (Ti) in precipitant condition, and more precisely the resistance against corrosion of Ti in presence of fluorides and the electrochemical interaction between Ti- amalgam couples in fluorinated solution. METHODS: The experimental test was made with the use of an electrochemical cell. The following materials were tested: commercially pure Ti and 2 types of amalgams, the Persistalloy (Prs) and the IQC. Palladium (IQC.P). The free corrosion potential of Ti and the amalgams, the polarization curves of both amalgams and the corrosion current of the Ti-amalgam couples in the measurements were performed in 3 different electrolytic solutions: Ringer solution, fluorinated neutral Ringer solution and acid fluorinated solution. The three corrosive media are described. RESULTS: The results showed that Ti could be damaged by the presence of fluorides with an acid pH: Ti potential becomes more negative in acid fluorinated solution. The corrosion currents between Ti and amalgam couples were considered: the amalgams underwent anodic oxidation in neutral Ringer, but a reversal phenomenon occurred in the fluorinated acid solution: Ti was damaged and the amalgams both Prs and IQC.P became the cathodic partner of the couple. In neutral fluorinated solution the IQC.P amalgam induced a significantly higher corrosion of Ti, when compared to the Prs one. CONCLUSION: Results clearly show the dependence of the Ti corrosion behaviour on the pH and composition of the solution and that the outcome of the damage is affected by the composition of other metals. PMID- 12783065 TI - [Squamous cell carcinoma of the lip. Retrospective analysis of 42 cases]. AB - BACKGROUND: To examine to what extent physicians follow a regional guidelines for the diagnosis, staging, treatment and follow-up of patients with squamous cell carcinoma of the lower and upper lip. METHODS: DESIGN: retrospective analysis of data from the medical records of 42 patients diagnosed with squamous cell carcinoma of the lower and upper lip during the period 1990-2000. SETTING: Department of head and neck pathology, oral cavity and audio-verbal communication of Naples University. PARTICIPANTS: the data were collected by this Department. RESULTS: In accordance with the TNM system, the 50% of cases was in stage T1, 33% in T2, 14% in T3. In few cases (3%) the lesion was in stage T4. All cases were in M0 and only 8% of cases presented nodal disease at diagnosis (N1). The type of treatment in relation to age was in agreement with the guidelines in 34% of cases. Of the 42 surgically-treated patients, only 4 showed, after 12 months, a local recurrence. CONCLUSIONS: Prognosis is good if squamous carcinoma is diagnosed early. PMID- 12783066 TI - [Desmoplastic ameloblastoma. Clinical and histopathological diagnostic criteria]. AB - A rare case of desmoplastic ameloblastoma observed in a 62-year old man presenting a swelling in the anterior mandible area with abnormal mobility of 33, 34, 43, 45, is described. Diagnosis of desmoplastic ameloblastoma was based on: anterior localization, radiopaque appearance, severe desmoplastic stroma and increasing infiltrate pattern of the tumor. The authors suggest to consider the desmoplastic histotype as a different nosological entity defined "desmoplastic ameloblastoma" since these clinical and histopathological features are missing in the other histologic variants of common ameloblastoma. A radical surgical treatment is suggested for this neoplasia. PMID- 12783067 TI - Modifier genes and heart failure. AB - Recent progress in genomic applications have led to a better understanding of the relationship between genetic background and cardiovascular diseases such as heart failure. A considerable component of the variability in heart failure outcome is due to modifier genes, i.e. genes that are not involve in the genesis of a disease but modify the severity of the phenotypic expression once the disease has developed. The strategy most commonly used to identify modifier genes is based on association studies between the severity of the phenotype of the disease (morbidity and/or mortality) and the sequence variation(s) of selected candidate gene(s). This strategy has showed that several polymorphisms of the beta1 and beta2 adrenergic receptors genes and the angiotensin converting enzyme gene are correlated to the prognosis of patients with heart failure. Recently, we have applied an experimental strategy, known as genome mapping, for the identification of heart failure modifier genes. Genome mapping has previously been used with success to identify the genes involved in the development of both monogenic and multifactorial diseases. We have showed that the prognosis of heart failure mice, induced through overexpressing calsequestrin, is linked to 2 Quantitative Trait Loci (QTL) localized on chromosome 2 and 3. Using both strategies (candidate gene and genome mapping) should allow us to identify a number of modifier genes that may provide a more rational approach to identify patients at risk for disease and response to therapy. PMID- 12783068 TI - B-type natriuretic peptides. A diagnostic breakthrough in heart failure. AB - B-type natriuretic peptide (BNP) is a neurohormone synthesized in the cardiac ventricles, which is released as N-terminal pro-brain natriuretic peptide (NT proBNP) and then enzymatically cleaved in to the NT fragment and the immunoreactive BNP. Both tests have been used to identify patients with congestive heart failure (CHF). Important considerations for these tests include their half-lives in plasma, dependence on renal function for clearance, and the interpretation of their units of measure. In general, a BNP level below 100 pg/mL has strong negative predictive value in the assessment of patients with dyspnea caused by a disorder other than CHF. In addition, BNP levels can be used to gauge the effect of short-term treatment of acutely decompensated heart failure, and the peptide has been shown to be a reliable independent predictor of sudden cardiac death. In the absence of renal dysfunction NT-proBNP has also been shown to be an independent predictor of sudden death in CHF patients. Because both a large area of myonecrosis or concomitant left ventricular failure are related to prognosis in acute coronary syndromes, B-type natriuretic peptides have also been linked to outcomes in this condition. This article describes the physiology and timing of release of B-type natriuretic peptides and the rationale for their use in the following settings: 1) evaluation of decompensated CHF, 2) screening for chronic CHF, 3) prognosis of CHF and sudden death, and 4) prognosis in acute coronary syndromes with inferred left ventricular dysfunction. PMID- 12783069 TI - Diastolic heart failure. Diagnosis, prognosis, treatment. AB - It is now abundantly clear that congestive heart failure caused by a predominant abnormality in diastolic function (i.e., diastolic heart failure) is common and causes a significant increase in morbidity and mortality. However, there is continued controversy surrounding the terminology used to describe patients with heart failure and the criteria used to make the diagnosis of diastolic heart failure. As a result, clinical therapeutic trials have been slow to develop and difficult to design. Fortunately, these controversies are yielding to an emerging consensus. Recent clinical studies have provided sufficient data to develop standardized diagnostic criteria to define diastolic heart failure. Experimental studies have provided increased insight into the mechanisms causing diastolic heart failure. Together, these clinical and experimental studies are being used to design targeted clinical trials to test effective treatments for diastolic heart failure. The purpose of this review is to describe the criteria used to diagnose diastolic heart failure, the effects of diastolic heart failure on prognosis, and approaches to treatment. PMID- 12783070 TI - Effects of ACE-inhibitors and beta-blockers on left ventricular remodeling in chronic heart failure. AB - In recent years, it has become increasingly recognised that a central feature of the disease progression associated with heart failure is the process of left ventricular remodeling. The remodeling process manifests as an increase in left ventricular volumes, leading to a rise in wall stress and a compensatory increase in myocardial mass. The left ventricle also gradually assumes a more spherical shape, resulting in functional mitral regurgitation leading to further haemodynamic overload, worsening myocardial function and an unfavourable clinical course. Accumulating clinical data support the hypothesis that the benefits in clinical outcome with ACE-inhibitors and beta-blockers may relate to modification of the remodeling process resulting in slowing of disease progression and preservation of contractile function. The general trend from a number of clinical studies indicates that whereas ACE-inhibitors seem to prevent progressive left ventricular dilatation, the third generation beta-blocker, carvedilol, may actually reverse the remodelling process by reducing left ventricular volumes and improving systolic function. Direct comparisons indicate that carvedilol has a similar safety and tolerability profile to ACE-inhibitors and thereby support the feasibility of administering this drug as first-line therapy in selected patients with mild to moderate chronic heart failure. Therefore, the decision to initiate treatment with carvedilol or an ACE-inhibitor might in future be tailored on an individual basis and followed thereafter by combination therapy at the earliest and safest opportunity. Finally, the possible development of treatment strategies addressing the cellular and molecular mechanisms responsible for the remodeling process and the recently published benefits of device therapies herald a combined, synergistic approach to the future management of heart failure. PMID- 12783071 TI - Aldosterone antagonism in addition to angiotensin-converting enzyme inhibitors in heart failure. AB - Although the role of the systemic renin-angiotensin-aldosterone system in the pathophysiology of heart failure is well-known for years, the impact of a local cardiac aldosterone system has been recognized recently. Aldosterone promotes cardiac hypertrophy and fibrosis in hypertension and heart failure and is involved in left ventricular remodeling after myocardial infarction. Plasma aldosterone levels in patients with heart failure are an indicator of a worse prognosis. Although ACE inhibitor therapy in these patients reduces plasma aldosterone levels, this effect is only transitory, a phenomenon referred to as "aldosterone escape". Even maximally recommended doses of ACE inhibitors do not completely prevent ACE-mediated formation of angiotensin II in chronic heart failure, and those patients with increased aldosterone levels during ACE inhibition have impaired exercise capacity. The RALES study has demonstrated convincingly that in patients with heart failure, addition of the mineralocorticoid receptor antagonist spironolactone (25 mg/d) to ACE inhibition markedly reduces mortality and prevents worsening heart failure. While reduction of excessive extracellular matrix turnover leading to decreased fibrosis appears to be the most important effect of spironolactone in heart failure, other mechanisms such as regression of hypertrophy, improvement of endothelial function, enhanced renal sodium excretion and antiarrhythmic actions may contribute. In RALES, low-dose spironolactone did not confer a substantial risk of hyperkalemia, however, with broader use of spironolactone in heart failure, cases of hyperkalemia associated with the use of this drug increase. Close control of serum potassium and creatinine and estimation of creatinine clearance are mandatory, especially in the presence of additional factors impairing renal function. The new and more selective aldosterone antagonist eplerenone which is devoid of some side effects of spironolactone, has been shown to be effective in hypertension and holds great promise as future therapeutic agent in patients with heart failure. PMID- 12783072 TI - [Risk of sudden death in heart failure patients]. AB - Sudden death is one of the more important cause of mortality in patients with chronic heart failure. The highest risk occurs among patients with less severe functional impairment, whereas patients in NYHA class IV usually die of progression of heart failure. Predictors of sudden death have been evaluated. Nevertheless, current methods of risk stratification for sudden death are still inadequate, especially in patients with advanced heart failure. Low left ventricular ejection fraction is widely used for the risk stratification, but it lacks of sensitivity and specificity in distinguishing patients with an increased arrhythmic mortality from those with an increased mortality due to pump failure. Unsustained ventricular tachycardia and inducibility at electrophysiological study may help identifying high-risk patients, requiring more aggressive therapy, as the ICD implantation. Heart rate variability and baroreflex sensitivity analysis have been utilized to obtain information on autonomic modulation, but with uncertain conclusion on the identification of high-risk patients. Increased QT dispersion, the presence of T-wave alternans and abnormal signal-averaged electrocardiography have also been proposed, but, up-to-now, any of these parameters showed a strong predictor power. In conclusion, our capability to identifying heart failure patients at risk for arrhythmic death is still far from being satisfactory. PMID- 12783073 TI - Heart failure. The pacing therapy. AB - Several medical therapies, including digoxin, angiotensin-converting enzyme inhibitors, and beta-blockers, have reduced the number of re-hospitalizations and slowed the progression of congestive heart failure (CHF) improving survival. Despite these benefits, medical therapy frequently fails to improve quality of life. Since 1990, there has been a growing interest in using cardiac pacing as additional treatment in severe CHF. Biventricular pacing is used in CHF patients with left bundle branch block (LBBB) to improve ventricular activation sequence which may lead to a more coordinated and efficient ventricular contraction. Since its introduction in CHF in 1994, biventricular pacing has been widely applied with many clinical trials and the development of new specific technology. With the development of new technology, the left ventricular catheterization via a coronary sinus vein, increased from 56% to over 95% during the last 2 years with an acceptable number of complications. Despite encouraging acute and short-term results, pacing strategies for CHF are still limited and currently regarded as investigational. It is clear that while some patients respond remarkably, this is high variable. Clinical trials are currently underway to assess the impact of cardiac resynchronization therapy on morbi-mortality and to assess the association with ventricular defibrillation. The whole validation process of cardiac resynchronization therapy should be completed on 2004-2005. Another novel mode of pacing therapy, which may be clinically appropriate for a broader range of CHF patients irrespective of the presence of LBBB, is contractility modulation, which involves sub-threshold pacing to increase intracellular calcium and enhance inotropy. PMID- 12783074 TI - Treatment of refractory congestive heart failure by cardiac resynchronization. AB - The contributions of long-term cardiac resynchronization as a supplemental treatment of refractory congestive heart failure have recently been reported. Several completed studies support the validity of this new therapy, capable of improving quality of life as well as increasing exercise capacity. These gains hinge on a careful patient selection, on the proper placement of the leads, particularly that responsible for left ventricular stimulation, and on an individualized patient follow-up. The results obtained thus fat fully justify considering cardiac resynchronization as an additional option in the treatment management of patients refractory to conventional measures. Ongoing studies should help to further defining its impact on morbidity and overall mortality, as well as the potential role for back up defribillator. PMID- 12783075 TI - Ongoing trials of cardiac resynchronisation. AB - Heart failure is an increasingly common and debilitating condition for which pharmacological therapy has, so far, provided only partial relief. Despite medical therapy the overall prognosis remains poor with high rates of sudden death and death from progressive heart failure. Device based therapies offer considerable promise both for the relief of symptoms and for improving prognosis. Cardiac resynchronisation therapy (CRT) has already been shown to improve the symptoms of heart failure when optimal pharmacological therapy (including aggressive diuretic therapy, ACE inhibitors, b-blockers and spironolactone) has failed. Two large trials (CARE-HF and COMPANION) are currently investigating the effects of CRT on morbidity and mortality in patients with heart failure and sinus rhythm who have left ventricular systolic dysfunction and ventricular dyssynchrony. A series of small and medium sized studies are assessing the effects of CRT in patients similar to the above but who also have atrial fibrillation. Other potential indications for CRT that are being explored include heart failure due to left ventricular diastolic function and for the prevention of iatrogenic dyssynchrony caused by conventional pacing. The MADIT-II study suggests a small benefit from routine implantation of defibrillators in patients with heart failure who have a markedly depressed (<30%) ejection fraction due to prior myocardial infarction even in the absence of specific marker of risk for sudden arrhythmic death. Much greater benefit was observed in patients with QRS >150 msec, an ECG marker for cardiac dyssynchrony. The COMPANION trial will not only assess the effects of CRT alone but also the effects of a combined CRT and defibrillator device. Premature over-interpretation of the limited amount of existing data threatens to undermine the evidence that will form the basis of future guidelines and funding decisions. Those involved in trials have an ethical duty to minimise device implantation into patients who have been randomised to the control group (cross-overs). Doctors may have difficulty explaining to patients why they implanted a CRT device should the current trials not show benefit. Patients should be warned that CRT is still an experimental therapy that has not yet been proven to alter outcome substantially. PMID- 12783076 TI - Experience with an ICD incorporating biventricular pacing. AB - Biventricular pacing for cardiac resynchronization is a promising therapy for symptomatic improvement in selected patients with underlying severe congestive heart failure. ICD treatment has been shown to prolong life in patients with life threatening ventricular tachyarrhythmias, but it does not improve quality of life. This review discusses current experience with ICD's incorporating biventricular pacing. PMID- 12783077 TI - Congestive heart failure. A new milestone for a better life. AB - Congestive heart failure (CHF) represents the 1st cause of death in the civil world. Despite considerable advances in the diagnosis and medical treatment of CHF, this condition remains a real "malignant" disease. The cardiac transplantation (CT) remains "gold standard" option for the treatment of patients with severe CHF in all age group; but only a small number of patients can receive it due to scarcity of donor organs. The increased successful clinical experience with the ventricular assist device (VAD) confirmed the indisputable importance of long-term mechanical circulatory support for patients who suffer acute hemodynamic deterioration. Over the past years, several miniaturized continuous flow VAD have been developed for clinical use. The authors report the data of worldwide and particularly Italian experience about middle and long-term pump performance in patients supported with axial-flow pump MicroMed DeBakey VAD in the bridge to cardiac transplantation setting. PMID- 12783078 TI - Cardiac transplantation of skeletal myoblasts for heart failure. AB - Heart failure has become the most prevalent cardiovascular syndrome, and its incidence continues to increase. Most cases of heart failure develop as a result of myocardial infarction. Although current treatment modalities have brought us the opportunity to reduce mortality and morbidity after myocardial infarction, our progress has plateaued due to our inability to treat the underlying problem, death of cardiomyocytes. Recently, a new option has emerged. Transplantation of undifferentiated cells into the damaged heart is a promising new treatment modality. These cells may have the capability of adapting to the cardiac environment, regenerating the damaged muscle, restoring cardiac function and preventing transition to heart failure. During the last few years many cell types have been proposed for cardiac repair and promising pre-clinical studies have moved some of these into the clinic. The most widely studied cell type is the progenitor cell of adult muscle, or the myoblast. When transplanted into the heart myoblasts are able to engraft and to a large degree regenerate the infarcted area. Although the feasibility of myoblast transplantation has been proven in animal models of infarction, many questions remain unanswered. In this review we will try to present an overview of where intracardiac myoblast transplantation stands and where it is heading. We also provide our insight into the future potential for myoblast transplantation clinically. PMID- 12783079 TI - Heart transplantation. An update and review. AB - Since the 1(st) heart transplant was performed over 30 y ago, recipients have benefited from dramatic improvements in survival and quality of life following the procedure. Approximately 3 500 people worldwide now undergo heart transplantation each year. The vast majority of patients will have no limitations in their daily activities and will remain free of re-hospitalization after the 1(st) year. The median survival following heart transplant is now 9 y, and it remains the treatment of choice for children and adults with end-stage heart disease despite emerging therapies. This article will review patient selection, perioperative management issues, medical management of recipients, and future trends in the treatment of this patient population. PMID- 12783080 TI - CT for thromboembolic disease. AB - Pulmonary embolism (PE) and deep venous thrombosis (DVT) constitute the two clinical manifestations of venous thromboembolic disease (VTE). The recent innovation of computed tomography venography (CTV) in conjunction with CT pulmonary arteriography (CTPA) provides a single noninvasive diagnostic test that can evaluate both components of VTE. PE is often an underestimated, underdiagnosed, and, consequently, undertreated disease entity. Herein, we review the epidemiology of thromboembolic disease, the diagnostic algorithm used in evaluation of patients with suspected VTE, and protocols for performing CTPA and CTV. Interpretation of these examinations is discussed in detail, because CTPA may pose new challenges to the practicing radiologist. PMID- 12783081 TI - MRI of shoulder instability: state of the art. PMID- 12783082 TI - Morgagni hernia: CT findings. PMID- 12783087 TI - Continuous intraarterial infusion of protease inhibitors in acute pancreatitis. AB - Low-molecular-weight protease inhibitors were synthesized and developed in Japan and are in clinical use there for the treatment of acute pancreatitis. However, protease inhibitors are not acknowledged as drugs for the treatment of pancreatitis in other countries. In a recent study in 30 patients with necrotizing pancreatitis, survival rate was improved (mortality rate 13.3%) by continuous intraarterial administration of low-molecular-weight protease inhibitors as compared to conventional treatment. In Italy it was reported that pancreatic disorder decreased after the administration of low-molecular-weight protease inhibitors before the start of endoscopic retrograde cholangiopancreatography. Low-molecular-weight protease inhibitors may be potential alternative drugs for the treatment and/or prevention of acute pancreatitis and, therefore, warrant further evaluation. (c) 2001 Prous Science. All rights reserved. PMID- 12783088 TI - Migraine headache treatment with eletriptan, a second-generation serotonin receptor agonist. AB - Eletriptan is a member of the triptan family of selective serotonin receptor agonists. These act against migraine by inducing vasoconstriction of the meningeal arteries. In pharmacological tests, eletriptan has shown high affinity for the 5-HT(1B/1D) receptors, which have been implicated in the etiology of migraine headache attacks. Pharmacokinetic evaluations have concluded that eletriptan offers greater bioavailability than sumatriptan, the effective predecessor to eletriptan. A rapid onset of action has also been characteristic of eletriptan in clinical trials, which have likewise demonstrated eletriptan's superiority to sumatriptan in granting relief of headache pain and other symptoms associated with migraine to a greater number of migraine patients. The drug has generally been well tolerated with only mild to moderate adverse events reported. These characteristics make eletriptan an attractive alternative to sumatriptan in the treatment of migraine. (c) 2001 Prous Science. All rights reserved. PMID- 12783086 TI - Dendritic-cell-specific ICAM3-grabbing non-integrin is essential for the productive infection of human dendritic cells by mosquito-cell-derived dengue viruses. AB - Dengue virus (DV) is a mosquito-borne flavivirus that causes haemorrhagic fever in humans. DV primarily targets immature dendritic cells (DCs) after a bite by an infected mosquito vector. Here, we analysed the interactions between DV and human monocyte-derived DCs at the level of virus entry. We show that the DC-specific ICAM3-grabbing non-integrin (DC-SIGN) molecule, a cell-surface, mannose-specific, C-type lectin, binds mosquito-cell-derived DVs and allows viral replication. Conclusive evidence for the involvement of DC-SIGN in DV infection was obtained by the inhibition of viral infection by anti-DC-SIGN antibodies and by the soluble tetrameric ectodomain of DC-SIGN. Our data show that DC-SIGN functions as a DV-binding lectin by interacting with the DV envelope glycoprotein. Mosquito cell-derived DVs may have differential infectivity for DC-SIGN-expressing cells. We suggest that the differential use of DC-SIGN by viral envelope glycoproteins may account for the immunopathogenesis of DVs. PMID- 12783089 TI - The participation of inflammatory cells in atherosclerosis. AB - Atherosclerosis is a slow progressive disease that is increasingly recognized to be a chronic inflammatory disease. Immune cells of various types are found directly in the atherosclerotic plaques at different stages of the disease. Most notable is the presence of macrophages and T lymphocytes, particularly in the beginning to intermediate stages of atherosclerotic development. Numerous investigators have shown that the participation of macrophages and T lymphocytes in the pathogenesis of atherosclerosis is important. However, partly due to the complex web of secreted mediators such as cytokines and growth factors that affects all aspects of the immunobiology of these cells and their surrounding cells, the exact roles that these cells play are still incompletely understood. Even less understood are the roles, if any, of other immune cells such as natural killer cells, neutrophils and mast cells, although information regarding these cells in atherosclerosis is rapidly emerging. (c) 2001 Prous Science. All rights reserved. PMID- 12783090 TI - The potential of selective COX-2 inhibitors in inflammatory and other diseases. AB - It has been known since the 1960s that prostaglandins play a key role in inflammation, fever and pain. In 1971, Sir John Vane discovered that the efficacy of nonsteroidal antiinflammatory drugs (NSAIDs) is derived by inhibiting a key enzyme in prostaglandin biosynthesis (1). This enzyme has subsequently been identified as cyclooxygenase (COX). Presently, there are two known COX isoforms with distinct physiological functions, COX-1 and COX-2. COX-1 is a constitutively expressed protein found in most tissues. In the gastric mucosa, COX-1 is involved in the production of cytoprotective prostacyclin. In platelets, COX-1 is required for the generation of thromboxane A(2), which promotes platelet aggregation leading to thrombus formation. In contrast to COX-1, COX-2 is strongly inducible by a variety of mitogens such as cytokines and growth factors, and plays a key role in the production of inflammatory prostaglandins. A William Harvey Research Conference on Defining the role of COX-2 inhibitors in inflammatory and other diseases in Porto, Portugal brought together 15 invited speakers to present their current findings on the clinical significance of COX-2 and strategies for selectively targeting this isoform. Information presented at the conference is summarized below. (c) 2001 Prous Science. All rights reserved. PMID- 12783091 TI - Amoxicillin-sulbactam: A combination with a sound pharmacokinetic-pharmacodynamic basis. PMID- 12783092 TI - An update of acute otitis media management: Experience with the amoxicillin sulbactam combination. PMID- 12783093 TI - Amoxicillin-sulbactam: A clinical and therapeutic review. AB - Combinations of beta-lactamase inhibitors with penicillins, especially aminopenicillins, have broad-spectrum antibacterial activity against most of the common pathogens of the respiratory and urinary tracts. This means that they are an ideal treatment for infections such as otitis media, sinusitis, special cases of pharyngeal tonsillitis (recurring forms, indirect pathogenic action, or after the failure of amoxicillin monotherapy), acute exacerbations of chronic bronchitis, cystitis, urethritis, etc. The amoxicillin-sulbactam combination is active against both beta-lactamase producer and nonproducer strains, and is effective against Gram-positive cocci (Streptococcus pneumoniae, Streptococcus pyogenes, Staphylococcus aureus of nonhospital origin), Gram-negative cocci (Neisseria gonorrhoeae, Moraxella catarrhalis and others), Gram-negative bacilli (nonhospital strains of Haemophilus influenzae, Escherichia coli and Klebsiella pneumoniae and others) and anaerobes. Its antimicrobial activity means that it is indicated in the treatment of respiratory, ear, nose and throat, urinary, dermatological and gynecological infections caused by susceptible germs, as well as in a variety of surgical situations (both as a treatment and as prophylaxis). It is absorbed very well orally, and its pharmacokinetic profile is very favorable, with very good tissue penetration. It is reasonably well tolerated: in a variable percentage of cases it may cause modification of intestinal transit and/or fecal consistency, which usually abates spontaneously. The new formulation for administration at intervals of 12 h is easier to use, is better tolerated and favors completion of therapy. In summary, the amoxicillin-sulbactam combination is effective and well tolerated in most infections of nonhospital origin in adults and children. (c) 2001 Prous Science. All rights reserved. PMID- 12783094 TI - Almotriptan in the treatment of migraine. AB - Almotriptan is a new selective serotonin 5-HT(1B/1D) receptor agonist which is chemically related to sumatriptan and is used in the acute treatment of migraine. Almotriptan, like the rest of the triptans, acts by inducing vasoconstriction of the meningeal arteries. The new drug has good oral bioavailability, and in clinical studies has been shown to be as effective or more effective than sumatriptan 100 mg in alleviating migraine headache and associated symptoms (nausea, vomiting, phonophobia and photophobia) when administered as a single oral or subcutaneous dose of 12.5 mg, this being the recommended dose. However, almotriptan has a very good tolerability profile, which has been shown to be superior to that of sumatriptan in a comparative trial. Therefore, almotriptan offers clear advantages over sumatriptan in the acute treatment of migraine attacks. (c) 2001 Prous Science. All rights reserved. PMID- 12783095 TI - Clinical use, therapeutic aspects and future potential of deferiprone in thalassemia and other conditions of iron and other metal toxicity. AB - The therapeutic aspects and future prospects of the new iron chelating drug deferiprone are reviewed, with an emphasis on its clinical use in thalassemia and other conditions of iron overload, imbalance and toxicity, as well as its possible use in other metal toxicity conditions. Orally administered deferiprone appears to be as effective as subcutaneous deferoxamine in the removal of iron in transfused iron loaded patients, with an equivalent therapeutic index profile in both animals and humans. Only about 10% of patients requiring iron chelation therapy worldwide receive deferoxamine mainly because of its high cost, toxicity and low compliance with subcutaneous administration. Deferiprone has been used by over 6000 patients in 40 countries worldwide, in some cases daily for more than 10 years, with very promising results. Doses of 50-120 mg/kg/day are effective in bringing patients to negative iron balance. Deferiprone increases urinary iron excretion, decreases serum ferritin levels and reduces liver iron in the majority of chronically transfused iron loaded patients. All of the toxic side effects of deferiprone are considered reversible and manageable, and include agranulocytosis, musculoskeletal and joint pains, gastrointestinal complaints and zinc deficiency. In general, the incidence of toxic side effects could be reduced by using lower doses or combination therapy with deferoxamine. The suggestion that deferiprone therapy may cause liver fibrosis has not been confirmed. New therapeutic protocols for maximizing the efficacy and minimizing the toxicity of deferiprone are being considered based on new findings in relation to its metal chelation, pharmacological, toxicological and metabolic properties. (c) 2001 Prous Science. All rights reserved. PMID- 12783096 TI - Indigenous drugs and atherosclerosis. AB - Coronary artery disease (CAD) is a major health problem worldwide and its increasing prevalence in younger age groups is of serious concern. Precipitating factors include changes in lifestyles, food fads and stress, as well as the impact of longevity resulting from the control and prevention of communicable diseases. Dietary practices can often lead to increased levels of lipids in blood or trigger an underlying disposition to atherosclerosis. Hence, dietary modification has been an important method for the management of CAD, with the aim of lowering lipid levels. However, long-term trials have shown that this approach reduces cholesterol and triglycerides only moderately. Drug therapy with clofibrate, cholestyramine, etc., has been successful but some disadvantages limit their long-term use. An alternative approach to the treatment of atherosclerosis has been the use of indigenous drugs. This article reviews some of the drugs of the traditional Indian system of medicine, Ayurveda, which have been shown to possess antiatherogenic properties. (c) 2001 Prous Science. All rights reserved. PMID- 12783097 TI - The role of n-3 polyunsaturated fatty acids in the prevention and treatment of cardiovascular disease. AB - Growing evidence has suggested an important role of n-3 polyunsaturated fatty acids in reducing risk of cardiovascular disease in the general population and patients with preexisting heart disease. In particular, several long-term epidemiologic studies have found an inverse association between fish consumption and risk of coronary heart disease or stroke. Two secondary prevention trials have found that increasing fish consumption or fish oil supplementation significantly reduced coronary death among patients with existing myocardial infarction. In addition, epidemiologic and clinical studies have suggested that alpha-linolenic acid (ALA), a short-chain n3-3 fatty acid from plant sources, may have similar cardiac benefits as long-chain n-3 fatty acids from fish. Potential mechanisms through which n-3 polyunsaturated fatty acids protect against CVD include their antiarrhythmic and antithrombotic effects, and improving insulin sensitivity and endothelial function. (c) 2001 Prous Science. All rights reserved. PMID- 12783098 TI - The role of taxanes in breast cancer treatment. AB - Cancer of the breast is a serious public health problem throughout the world. Chemotherapy remains the mainstay of treatment of both early stage and metastatic breast cancer. New advances in chemotherapy include paclitaxel and docetaxel. This article reviews the historical background to the introduction of these highly active agents, their biochemistry, mechanism of action and pharmacology, dose and schedule in the treatment of breast cancer, first- and second-line use as well as anthracycline/taxane combinations. Adjuvant, neoadjuvant use and the emergence of weekly taxanes is discussed. (c) 2001 Prous Science. All rights reserved. PMID- 12783099 TI - Therapeutic angiogenesis with gene transfection for ischemic heart disease. AB - Recent advances in modern medicine have made it possible to understand disease processes at the genetic level and to apply this knowledge to new treatment strategies. Genetic engineering studies conducted in the field of cardiovascular medicine have led to the use of gene therapy as a new means for treating ischemic heart disease. The incidence of ischemic heart disease, with underlying conditions of diabetes mellitus and arteriosclerosis, has increased in recent years and is a major cause of myocardial infarction, cerebrovascular disease and death. Patients with these diseases often do not respond satisfactorily to conventional treatments. Consequently attention has turned to the area of revascularization therapy utilizing the introduction of genes. (c) 2001 Prous Science. All rights reserved. PMID- 12783100 TI - Current and future therapies for ischemic cerebrovascular disease. II. Surgery and new treatments on the horizon. AB - Stroke is the third leading cause of death in the adult population. Numerous neuroprotective agents and procedures have been developed and a new wave of therapies is now on the horizon with the potential to minimize ischemic brain damage. On the other hand, surgical treatment has also played an important role in the treatment of stroke. This article highlights recent advances in stroke treatment, including surgical and neurointerventional radiological procedures, as well as potential new therapies. (c) 2001 Prous Science. All rights reserved. PMID- 12783101 TI - Fecal calprotectin as an index of intestinal inflammation. AB - The assessment of inflammatory activity in intestinal disease in man can be done using a variety of different techniques, from measurement of conventional noninvasive acute-phase inflammatory markers in plasma (C-reactive protein and the erythrocyte sedimentation rate) to the direct assessment of disease activity by intestinal biopsy. However, most of these techniques have significant limitations when it comes to assessing functional components of the disease that relate to activity and prognosis. Here we briefly review the value of a novel emerging intestinal function test, fecal calprotectin. Single stool assay of neutrophil-specific proteins (calprotectin, lactoferrin) give the same quantitative data on intestinal inflammation as the 4-day fecal excretion of indium-111-labeled white cells. Elevated levels of fecal calprotectin have been demonstrated in patients with NSAID-induced enteropathy and have been used in the diagnosis of colorectal cancer. Fecal calprotectin is increased in over 95% of patients with inflammatory bowel disease (IBD) and correlates with clinical disease activity. It reliably differentiates between patients with IBD and irritable bowel syndrome (IBS). More importantly, at a given fecal calprotectin concentration in patients with quiescent IBD, the test has a specificity and sensitivity in excess of 85% in predicting clinical relapse of disease. This suggests that relapse of IBD is closely related to the degree of intestinal inflammation and suggests that targeted treatment at an asymptomatic stage of the disease may be indicated. (c) 2001 Prous Science. All rights reserved. PMID- 12783102 TI - Tardive dyskinesia: An update. AB - Tardive dyskinesia (TD) is a serious motor side effect of chronic neuroleptic therapy. TD is a complex hyperkinetic syndrome consisting of choreiform, athetoid or rhythmic abnormal involuntary movements. The face, mouth and tongue are most frequently involved (orofacial type), but a variety of less frequent motor abnormalities of the upper and lower limbs and of the trunk may also occur. TD usually has a delayed onset and the intensity of the syndrome may fluctuate over time. The most serious aspect of TD is that it may persist for months or years after drug withdrawal and in some patients is irreversible. In spite of the prevalence and known etiology that chronic neuroleptic treatment causes TD, relatively little is known about the primary pathological mechanism underlying the condition. Abnormalities in various neurotransmitter systems have been implicated in the pathophysiology of TD, including the dopaminergic, GABAergic, serotonergic and noradrenergic systems. Recently, excitotoxicity of the glutamatergic system and oxidative stress have received much attention. Three general types of animal models have contributed to our knowledge of TD and can be described as homologous, analogous and correlational models. There are no empirically validated guidelines to follow when choosing a suppressive agent. In general, therapeutic trials have attempted to manipulate the dopaminergic, GABAergic, serotonergic and noradrenergic systems, in part due to theories on the pathophysiology of TD. None of these medications has proven successful in the majority of patients. Much more research is needed in order to increase our understanding of TD and to develop better therapeutics for its treatment. (c) 2001 Prous Science. All rights reserved. PMID- 12783103 TI - The role of biological therapy in inflammatory bowel disease. AB - Substantial evidence suggests a central role for TNF-alpha in the pathogenesis of IBD. This molecular observation has been supported by clinical trials with anti TNF therapies. The most extensively investigated among the various anti-TNF agents is infliximab. Clinical trials to date have demonstrated its efficacy in inducing remission in patients with moderately active, refractory Crohn's disease (CD) and in managing patients with CD complicated by fistulas. One advantage of infliximab is its rapid onset of action. However, as expected with most medications used to treat patients with IBD, the effect of infliximab is of limited duration, with the response lasting 2-3 months in most patients. The efficacy of repeated infusions of infliximab in maintaining remission in patients with inflammatory CD has been demonstrated in one trial to date. The results from the ACCENT I trial should soon be available. Many other important questions regarding the use of infliximab remain unanswered. These include the optimal schedules of infusions, the effect of concomitant therapy with aminosalicylates, immunomodulators and antibiotics, and the timing and indication of using infliximab in the general management algorithm of a patient with CD. Certainly, the efficacy of infliximab in the treatment of ulcerative colitis (UC) remains to be further explored in a controlled fashion, though preliminary uncontrolled data suggests efficacy. As experience with infliximab use accumulates, more data will become available regarding its safety with either short-term or long-term use. A large body of evidence exists regarding the short-term safety of infliximab. The concern of increased risk of hypersensitivity-like reactions with longer interval between treatments will also need to be addressed. The currently available data supports that infliximab is safe and well tolerated. Other anti-TNF therapies will also need to be investigated with the same rigor before widespread use can be advocated. In addition to these agents, advances in molecular engineering techniques have further expanded the array of biologic therapies available to treat IBD. These newer therapies hold promise in targeting specific pathways of the pathogenesis of IBD that may be different from all prior therapies. Certainly, the anti-TNF therapies and others aforementioned have taken the field of IBD into a new and exciting generation, the biological era. (c) 2001 Prous Science. All rights reserved. PMID- 12783104 TI - Voriconazole: A second-generation triazole. AB - Voriconazole, a derivative of fluconazole, is an antifungal triazole with expanded spectrum of activity against a variety of yeasts and filamentous fungi. It exhibits fungicidal activity against Aspergillus species. Preliminary data from animal models and clinical trials show promise in the treatment of candidiasis and aspergillosis. Of concern is the development of resistance to voriconazole and cross-resistance with other azoles. Voriconazole is a welcome addition to our limited armamentarium against potentially fatal fungal infections. (c) 2001 Prous Science. All rights reserved. PMID- 12783105 TI - Good breeding matters: in-bred rodents provide genetic insights into platelet secretion. PMID- 12783106 TI - Enoxaparin monotherapy without oral anticoagulation to treat acute symptomatic pulmonary embolism. AB - Conventional anticoagulation for symptomatic pulmonary embolism consists of continuous intravenous unfractionated heparin as a "bridge" to oral anticoagulation. This strategy requires 5 days or more of intravenous heparin while oral vitamin K antagonists gradually achieve a therapeutic effect. Oral vitamin K antagonists require frequent blood testing to optimize dosing, and their interactions with other medications and foods make regulation difficult. Therefore we tested a different approach to therapy: long-term enoxaparin monotherapy. We randomized 60 symptomatic pulmonary embolism patients in a 2:1 ratio to 90 days of enoxaparin as monotherapy without warfarin (N=40) or to intravenous unfractionated heparin as a "bridge" to warfarin, target INR 2.0-3.0 (N=20). Enoxaparin patients received 1 mg/kg twice daily for 14 days during the acute phase followed by randomized assignment during the chronic phase to 1.0 mg/kg vs. 1.5 mg/kg once daily. In an intention-to-treat analysis, 3 of the 40 enoxaparin patients developed recurrent venous thromboembolism compared with 0 of 20 standard therapy patients (p = 0.54). One of the 40 enoxaparin patients had a major hemorrhagic complication compared with 2 of the 20 standard therapy patients (p = 0.26). Median hospital length of stay was shorter with enoxaparin compared to standard therapy (4 vs. 6 days) (p = 0.001). Following our study we can conclude that extended 3-month treatment with enoxaparin as monotherapy for symptomatic, acute pulmonary embolism is feasible and warrants further study in a large clinical trial. PMID- 12783107 TI - Posttranscriptional regulation of PAI-1 gene expression. AB - The plasminogen activator-plasmin cascade is involved in multiple physiological and pathological processes including fibrinolysis, wound healing, fibrosis, angiogenesis, embryo implantation and tumor cell invasion and metastasis. Plasminogen activator-inhibitor type 1 (PAI-1) is the major physiological regulator of plasminogen activation. PAI-1 is expressed in a variety of mammalian cells and is regulated by growth factors, cytokines and hormones, including agents that elevate cAMP levels. Although cyclic nucleotide regulation of PAI-1 is observed in diverse cell types in various species, including human, limited studies have addressed the mechanism of this regulation. Here we review our work on the regulation of PAI-1 mRNA degradation in HTC rat hepatoma cells, describing the cis-acting cAMP-responsive sequence in the transcript and a novel RNA binding protein that interacts with it. Potential mechanisms by which this RNA-binding protein may be involved in cyclic nucleotide regulation of mRNA stability are discussed and cAMP regulation of PAI-1 in other systems is summarized. PMID- 12783108 TI - Viral hemorrhagic fever--a vascular disease? AB - The syndrome of "viral hemorrhagic fever" in man caused by certain viruses, such as Ebola, Lassa, Dengue, and Crimean-Congo hemorrhagic fever viruses, is often associated with a shock syndrome of undetermined pathogenesis. However, the vascular system, particularly the vascular endothelium, seems to be directly and indirectly targeted by all these viruses. Here we briefly summarize the current knowledge on Marburg and Ebola virus infections, the prototype viral hemorrhagic fever agents, and formulate a working hypothesis for the pathogenesis of viral hemorrhagic fever. Infections with filoviruses show lethality up to 89% and in severe cases lead to a shock syndrome associated with hypotension, coagulation disorders and an imbalance of fluid distribution between the intravascular and extravascular tissue space. The primary target cells for filoviruses are mononuclear phagocytotic cells which are activated upon infection and release certain cytokines and chemokines. These mediators indirectly target the endothelium and are thought to play a key role in the pathogenesis of filoviral hemorrhagic fever. In addition, direct infection and subsequent destruction of endothelial cells might contribute to the pathogenesis. Filoviruses, particularly Ebola virus, encode nonstructural glycoproteins which are released from infected host cells. Their function as potential determinants in pathogenicity remains to be investigated. PMID- 12783109 TI - Anti-hirudin antibodies alter pharmacokinetics and pharmacodynamics of recombinant hirudin. AB - Recombinant hirudin (r-hirudin) is a potent direct thrombin inhibitor with immunogenic properties. Anti-hirudin antibodies (aHAb) are detected in up to 74% of patients treated with r-hirudin for more than 5 days. aHAb may alter the pharmacokinetics and pharmacodynamics of r-hirudin. The effects of aHAb on the pharmacokinetics of r-hirudin were investigated in rats receiving r-hirudin intravenously either without aHAb (controls), 15 min after intravenous administration of non-specific antibodies or aHAb, and after pre-incubation with aHAb. When both were compared to controls and pre-treatment with non-specific antibodies, aHAb significantly altered the pharmacokinetics of r-hirudin with similar effects in both approaches: In the presence of aHAb, the volume of distribution in a steady state and total plasma clearance were diminished, while the half-life of elimination was prolonged. Both the maximum r-hirudin plasma concentration and the area under the curve were increased. In addition, r-hirudin filtration by high-flux hemodialyzer membranes (polysulfone, AN69) was investigated 1) in the absence of aHAb, 2) in the presence of non-specific mouse antibodies, and 3) in the presence of three monoclonal aHAb. In the absence of aHAb, both hemodialyzers allowed for significant r-hirudin filtration. Non specific mouse antibodies did not markedly affect r-hirudin filtration. By contrast, all three aHAb almost completely hindered r-hirudin filtration. aHAb varied in their capacity to neutralize r-hirudin. In conclusion, aHAb markedly alter the pharmacokinetics of r-hirudin leading to r-hirudin accumulation. In the presence of aHAb, hemofiltration does not allow for rapid reduction of r-hirudin concentration. aHAb are capable of modifying pharmacodynamics of r-hirudin. Close monitoring of aHAb-positive patients treated with r-hirudin is considered mandatory. PMID- 12783110 TI - A FV multiallelic marker detects genetic components of APC resistance contributing to venous thromboembolism in FV Leiden carriers. AB - Activated protein C resistance (APCR) is a major risk factor for venous thromboembolism (VTE). Although the factor V (FV) Leiden mutation accounts for the vast majority of APCR cases, other polymorphisms may contribute to the APCR phenotype. Genetic components of APCR and thrombophilia were investigated by two dinucleotide repeats, characterized in introns 2 and 11 of the FV gene. Only the intron 11 marker was genetically stable and thus suitable for further analysis. Its allelic frequencies were found to differ significantly (P=0.003) between subjects selected for increased APCR in the absence of the FV R506Q mutation (n=70, normalized ratios /=1.31). Genotype differences were also found (P=0.017) between FV R506Q heterozygotes (n=100) who had experienced previous VTE and those (n=100), who were still asymptomatic for VTE. Significance was mostly driven by the relative over-representation of the 12R allele and to a minor extent by the under-representation of the 15R allele among the symptomatic versus the asymptomatic FV Leiden carriers. Two SNPs (4070A/G and 2391A/G) were found to underlie the 12R and 15R alleles respectively, and marked extended haplo-types, previously (HR2) or newly (HT2) identified. Only the FV HR2 differed (P=0.002) in frequency between the two groups of FV R506Q heterozygotes, suggesting that it represents the most relevant FV genetic component of APCR or VTE detectable by this experimental and clinical approach. Our analysis indicates that frequent FV genetic components might contribute to shape the risk for VTE in FV Leiden carriers. PMID- 12783111 TI - Effects of antioxidant vitamins C and E on endothelial function and thrombosis/fibrinolysis system in smokers. AB - Smoking is associated with endothelial dysfunction and abnormalities in thrombosis/fibrinolysis system, possibly through increased oxidative stress. In this study we investigated the effect of combined antioxidant treatment with vitamins C and E on endothelial function and plasma levels of plasminogen activator inhibitor (PAI-1), von Willebrand factor (vWF), tissue plasminogen activator (tPA) and factor VII (fVII), in smokers. Forty-one healthy smokers were randomly divided into 4 groups receiving vitamin C 2g/day (group A), vitamin C 2g/day plus vitamin E 400 IU/day (group B), vitamin C 2g/day plus vitamin E 800 IU/day (group C) or no antioxidants (controls, group D), for 4 weeks. Forearm blood flow was measured using venous occlusion strain-gauge plethysmography. Forearm vasodilatory response to reactive hyperemia (RH%) or to sublingual nitroglycerin administration (NTG%) were considered as indexes of endothelium dependent or independent dilation respectively. After treatment, RH% was increased only in groups B (p <0.05) and C (p <0.001) but not in groups A and D. Plasma levels of PAI-1 and vWF were decreased only in group C (p <0.05 for both), while PAI-1/tPA ratio was significantly decreased in both groups B and C (p <0.05 for both). NTG% and plasma levels of tPA and fVII remained invariable in all groups. In conclusion, combined administration of vitamin C and vitamin E at high dosages, improved endothelial function and decreased plasma levels of PAI-1, vWF and PAI-1/tPA ratio in chronic smokers. PMID- 12783112 TI - A new monoclonal antibody, mAb 204-11, that influences the binding of platelet GPVI to fibrous collagen. AB - The newly identified platelet collagen receptor glycoprotein VI binds to fibrous collagen, inducing platelet activation. Several antibodies against GPVI have been reported, including a patient's auto-antibodies, that activates platelets through their ability to crosslink this glycoprotein. We have developed a monoclonal antibody (mAb) against GPVI using the recombinant extracellular domain of GPVI as an antigen. This antibody, mAb 204-11, induced platelet aggregation and tyrosine phosphorylation of proteins similar to those induced by GPVI-reactive proteins, collagen and convulxin. Its interaction with GPVI was analyzed by measuring the effect of the antibody on GPVI binding to collagen using a dimeric form of recombinant GPVI, GPVI-Fc2. MAb 204-11 inhibited the binding of GPVI-Fc2 to fibrous collagen particles, but enhanced the GPVI binding to immobilized collagen, suggesting that the antibody binds to a region near the collagen binding site of GPVI. MAb 204-11 also inhibited the GPVI binding to convulxin at a low concentration, but not completely. Since mAb 204-11 reacts specifically with GPVI and is applicable for immunoblotting and immunoprecipitation, this antibody would be useful for studies on GPVI. PMID- 12783113 TI - A rapid, automated flow cytometric method to measure activated degranulated platelets by density determination. AB - Platelet activation is reported to correlate with acute coronary syndromes. A platelet analysis method on the ADVIA 120 Hematology System provides rapid analysis of platelet density, reported as mean platelet component (MPC) concentration, utilizes routine hematology specimens, requires no pre-treatment, and thirty seconds to generate results. Sub-populations of platelets separated by density gradients showed excellent correlation with the ADVIA 120 MPC parameter (r = 0.997). Platelet activation induced by thrombin treatment resulted in a shift of platelets into the lowest density fraction (d15 mmHg). In CHF rats, ACE inhibition significantly reduced platelet P-selectin expression while bound fibrinogen was not modulated. Eplerenone reduced P-selectin expression to a comparable extent, while platelet-bound fibrinogen was normalised. Combination therapy with eplerenone and trandolapril completely abolished both the increased P-selectin expression as well as fibrinogen binding. Phosphorylation of platelet vasodilator stimulated phosphoprotein (VASP) at both Ser(157) and Ser(239), which reflects the activity of platelet inhibitors including nitric oxide, was significantly reduced in platelets from placebo-treated CHF rats, and was completely normalised by combination treatment, but only marginally increased by either mono-therapy. The results show that platelet activation was evident only in CHF rats. Monotherapy with ACE inhibition or eplerenone partially reduced this increased platelet activation, which was completely rescued to basal levels by combination therapy. Increased nitric oxide bioavailability can only partially explain the reduced platelet activation by eplerenone and ACE inhibition. PMID- 12783116 TI - Genetic mapping and characterization of the bleeding disorder in the fawn-hooded hypertensive rat. AB - Release of platelet dense granule contents occurs in response to vascular injury, playing an important role in platelet aggregation and primary hemostasis. Abnormalities of the platelet dense granules results in a bleeding disorder of variable severity termed "storage pool defect" (SPD). We have examined the fawn hooded hypertensive (FHH) rat as a model of SPD in order to genetically map the locus (Bd) responsible for prolonged bleeding. Platelet function assays of the FHH rat confirmed the presence of a platelet dense granule SPD. However electron microscopy and lysosomal enzyme assays indicated differences between the FHH rat and other rodent models of SPD. Genetic mapping through the use of congenic FHH rats localized the Bd locus to an approximately 1 cM region on rat chromosome 1. Through the use of comparative mapping between species and analysis of the initial draft of the rat genome assembly, six known and thirty-four putative genes were identified in the Bd locus. None of these genes have been previously implicated in platelet function. Therefore positional cloning of the gene responsible for the bleeding disorder in the FHH rat will lead to new insights in platelet physiology, with implications for diagnosis and management of hemostatic and thrombotic disorders. PMID- 12783117 TI - Intercellular adhesion molecule-1 (ICAM-1) expression is upregulated by thrombin in human monocytes and THP-1 cells in vitro and in pregnant subjects in vivo. AB - Monocytes play a pivotal role in both the inflammatory and coagulation responses, which may be mediated through a variety of adhesion molecules on the cell surface, including intercellular adhesion molecule-1 (ICAM-1). Monocytes also possess thrombin receptors. In the current study, we have demonstrated that thrombin can upregulate ICAM-1 mRNA and induce ICAM-1 expression on the monocyte in vitro and that, in vivo, higher monocyte ICAM-1 expression is observed in pregnancy (which is characterised by a physiological increase in thrombin generation). In pregnant subjects, a positive correlation between monocyte ICAM-1 expression and a number of markers of vascular/thrombotic disease (including blood group, acquired activated protein C resistance and non-fasting plasma triglyceride levels) was observed. We also observed a significant relationship between monocyte ICAM-1 expression and soluble plasma ICAM-1 levels, which would be consistent with a contribution of monocytic ICAM-1 to the levels of free ICAM 1 observed in plasma during pregnancy. Consistent with a role in fibrinogen binding, our preliminary in vivo results suggest that monocyte ICAM-1 expression may be a useful marker of the thrombotic/inflammatory response, although further work is required to assess the relationship of monocyte ICAM-1 expression in thrombotic disorders. PMID- 12783118 TI - Distinct accumulation patterns of soluble forms of E-selectin, VCAM-1 and ICAM-1 upon infusion of TNFalpha in tumor patients. AB - The transmigration of leukocytes across the endothelium is a prerequisite for the inflammatory process. Leukocyte-endothelium interaction is regulated by several endothelial adhesion molecules, such as intercellular adhesion molecule-1 (ICAM 1), vascular cell adhesion molecule-1 (VCAM-1) and E-selectin. Their expression is enhanced by inflammatory mediators, such as TNalpha. In vivo a small part of these adhesion molecules is shed by proteases, and can be detected in the circulation. In this study, the time course of the TNalpha-induced accumulation in the blood of three circulating soluble adhesion molecules, sE-selectin, sICAM 1 and sVCAM-1, was studied in plasma samples of tumor patients enrolled in a phase I trial receiving TNalpha. Two different cohorts were studied. Eleven patients received a continuous 24-hour TNalpha infusion while 10 other patients received a 5-day continuous TNalpha infusion. After 24 hours of TNalpha infusion sE-selectin levels increased by 1985 +/- 312 %, sVCAM-1 by 301 +/- 19 % and sICAM 1 by 445 +/- 82 %. Differences in accumulation patterns were observed after 5 days of continuous TNalpha infusion. sVCAM-1 and sICAM-1 levels showed an increase during the infusion with a maximum at 3 to 5 days and stayed elevated after discontinuation of the TNalpha infusion. In contrast, sE-selectin reached its peak at day 1 and declined thereafter. In conclusion, sVCAM-1 and sICAM-1 show a different accumulation pattern upon TNalpha infusion as compared to sE selectin in man. PMID- 12783119 TI - Increased inflammatory status and higher prevalence of three-vessel coronary artery disease in patients with concomitant coronary and peripheral atherosclerosis. AB - The aim of this study was to determine whether patients with coronary artery disease (CAD) and concomitant peripheral arterial disease (PAD) have a greater inflammatory status than those with CAD alone. To this aim, we evaluated PAD (ankle/brachial pressure index <0.9), and measured plasma levels of C-reactive protein (CRP), interleukin-6 (IL-6) and the soluble forms of intercellular adhesion molecule-1 (sICAM-1) and vascular cell adhesion molecule-1 (sVCAM-1) in 234 patients who underwent coronary angiography. Median levels of CRP, IL-6 and sICAM-1 were higher in the CAD without PAD (n=134) and CAD+PAD (n=40) groups than in 60 patients without either disease ("controls"). Median CRP values were higher in patients with CAD+PAD than in patients with CAD alone (4.7 mg/L [1.5; 7.6] vs 2.4 mg/L [0.9; 3.8], p < 0.01).Three-vessel CAD was diagnosed in 60% of CAD+PAD patients and in 21% (p< 0.01) of CAD only patients. After adjustment for confounding factors, only PAD was independently associated with three-vessel CAD (p<0.001). This association was maintained after adjustment for IL-6, the only inflammatory parameter significantly associated with three-vessel CAD at univariate analysis (p<0.01). In conclusion, in CAD the coexistence of PAD is associated with a greater inflammatory status and more widespread coronary atherosclerosis. These results could help to explain the high cardiovascular risk of patients with concomitant CAD and PAD and suggest that PAD be included among the variables used to identify CAD patients for further diagnostic evaluation. PMID- 12783120 TI - PAI-1 level and the PAI-1 4G/5G polymorphism in relation to risk of non-fatal myocardial infarction: results from the Stockholm Heart Epidemiology Program (SHEEP). AB - This study examines the relationship between plasma Plasminogen Activator Inhibitor-1 (PAI-1) activity and the PAI-1 4G/5G polymorphism, and their association with the risk of myocardial infarction (MI). Furthermore, this study explores whether a high level of PAI-1 or whether the PAI-1 4G/5G polymorphism synergistically interacts with any established environmental risk factor for MI. This case-referent study of subjects aged 45-70 and living in Stockholm includes 851 men and 361 women with first-time MI and 1051 men and 505 women who were randomly chosen as referents from a population register. The adjusted odds ratio (OR) of MI was 1.9 (95% confidence interval [CI] 1.4-2.8) for men with a plasma PAI-1 activity level greater than the 90 th percentile value of referents. The corresponding OR for women was 1.5 (95% CI 0.9-2.5). A strong indication of synergistic interaction was found in men for the co-exposure to high plasma PAI-1 activity and current smoking, an adjusted synergy index score of 3.9 (95% CI 1.2 13.2). In women, the 4G allele was associated slightly with an increased risk of MI, OR 1.4 (95% CI 1.0-1.9). This association was not found in men. There were no clear indications of synergistic interaction effects involving the PAI-1 4G/5G polymorphism and the environmental exposures considered (cigarette smoking, physical inactivity, overweight, diabetes mellitus, hypercholesterolaemia, hypertension, high C-reactive protein and hypertriglyceridaemia). PMID- 12783121 TI - Recombinant human tissue plasminogen activator protects the basal lamina in experimental focal cerebral ischemia. AB - While recombinant tissue plasminogen activator (rt-PA) is successfully used in human ischemic stroke, it may also cause hemorrhagic complications. Animal experiments have shown that hemorrhages are related to microvascular basal lamina damage. We investigated the effects of different doses of rt-PA on the brain microvasculature. Experimental cerebral ischemia in rats was induced for 3 h and followed by 24 h reperfusion (suture model). Each group of rats (n = 6) received either treatment (0.9, 9, or 18 mg rt-PA/kg body weight) or saline (control group) at the end of ischemia. The loss of microvascular basal lamina antigen collagen type IV was measured by Western blot of the ischemic and non-ischemic basal ganglia and cortex. Compared with the contralateral non-ischemic area, collagen type IV was significantly reduced in the ischemic area: (basal ganglia/cortex) 43% +/- 9% / 64% +/- 4 %. Low/moderate doses of rt-PA had a protective effect: 0.9 mg 79% +/- 3% / 89% +/- 6%, 9 mg 72% +/- 9%/ 81% +/- 12% (p < 0.05). Higher doses of rt-PA (18 mg) had a similar effect as seen in untreated controls: 57% +/- 11% / 59% +/- 9% (p < 0.05, Anova). MMP-9 and MMP-2, measured by gelatine zymography, steadily increased over higher doses of rt-PA: MMP-9 (basal ganglia/cortex): control 115% +/- 4% / 123% +/- 3% compared with 18 mg rt-PA 146% +/- 5%/ 162% +/- 6% (p < 0.05) and MMP-2: control 109% +/- 4%/ 116% +/- 5% and 18 mg rt-PA 222% +/- 15%/ 252% +/- 2% (p < 0.05). Low to moderate doses of rt-PA protect the microvascular basal lamina, whereas high doses of rt PA have the opposite effect, probably due to increased coactivation of MMP-2 and MMP-9. PMID- 12783123 TI - Fibrinogen mediates bladder cancer cell migration in an ICAM-1-dependent pathway. AB - Fibrinogen (fg), present in tumor matrices, has been demonstrated to be determinant in metastatic potential. We have recently shown that fg/ICAM-1 interactions are involved in leukocyte migration across endothelial cell monolayers. Using bladder transitional cell carcinoma as a model, we will show in this study that bladder high grade tumor cell lines express ICAM-1, and that this expression induces an fg-mediated migration. This phenomenon was dependent on ICAM-1/fg interaction as well as RhoA activity. ICAM-1 was concentrated in focal adhesion plaques when tumor cells were allowed to adhere on immobilized fg, suggesting a role in cell migration. The addition of fg induced a 3- to 6-fold enhancement of bladder tumor cell migration through HUVEC monolayers. This process was inhibited by an anti-ICAM-1 antibody blocking fg binding, demonstrating that ICAM-1/fg interaction was involved in the extravasation process. Finally, immunohistological studies revealed that the expression of ICAM 1 was closely associated with an infiltrative histological phenotype. PMID- 12783122 TI - Expression of BCL2L12, a new member of apoptosis-related genes, in breast tumors. AB - Apoptosis, a normal physiological form of cell death, is critically involved in the regulation of cellular homeostasis. If the delicate balance between cell death and cell proliferation is altered by a defect in the normal regulation of apoptosis signaling, a cell population is able to survive and accumulate, thereby favoring the acquisition of further genetic alterations and promoting tumorigenesis. Dysregulation of programmed cell death mechanisms plays an important role in the pathogenesis and progression of breast cancer, as well as in the responses of tumors to therapeutic intervention. Overexpression of anti apoptotic members of the Bcl-2 family such as Bcl-2 and Bcl-XL has been implicated in cancer chemoresistance, whereas high levels of pro-apoptotic proteins such as Bax promote apoptosis and sensitize tumor cells to various anticancer therapies. Recently, a new member of the Bcl-2 family, BCL2L12, was cloned. The BCL2L12 gene is constitutively expressed in many tissues, suggesting that the encoded protein serves an important function in different cell types. In the present study, the expression of BCL2L12 gene was analyzed by reverse transcription-PCR (PT-PCR) in 70 breast cancer tissues. Our results indicate that BCL2L12 positive breast tumors are mainly of lower stage (I/II) or grade (I/II) (p=0.02 or p=0.04 respectively). Cox regression analysis revealed that BCL2L12 expression is positively related to disease-free (DFS) and overall survival (OS) at both univariate and multivariate analysis (p=0.021, p=0.029, p=0.032, p=0.044 respectively). Kaplan-Meier survival curves also demonstrated that patients with BCL2L12-positive tumors have significantly longer DFS and OS (p=0.002 and p<0.001 respectively). BCL2L12 expression may be regarded as a new independent favorable prognostic marker for breast cancer. PMID- 12783124 TI - Impact of pretreatment thrombocytosis on survival in primary breast cancer. AB - Platelet count has been reported to have predictive value in various cancer entities. In the case of breast cancer, evidence about involvement of platelets is still incomplete. Our objective was to assess the influence of pretreatment thrombocytosis on survival and establish its prognostic relevance for breast cancer patients. We performed a retrospective, multivariate analysis of 4,300 patients with early-stage breast cancer. All subjects participated in one of five prospective, randomized, multicenter trials conducted by the Austrian Breast and Colorectal Cancer Study Group. Thrombocytosis was defined as a platelet count exceeding 400 G/L. Median follow-up was 52 months. Univariate and multiple Cox regression models were calculated for overall survival (OS), breast cancer related survival and disease-free survival (DFS). Pretreatment thrombocytosis was observed in 161 patients (3.7%). Estimated median OS, breast cancer-related survival and DFS for patients with versus those without thrombocytosis was 71.0 versus 99.5, 72.0 versus 100.9, and 80.4 versus 88.4 months, respectively (p = 0.0054, p = 0.0095, p = 0.0199). A multiple Cox regression model including tumor and nodal status, grading, age, hormone receptor status and pretreatment thrombocytosis identified pretreatment thrombocytosis as an independent predictive factor for OS (p = 0.0064) and breast cancer-related survival (p = 0.0162). Multivariate analysis failed to identify pretreatment thrombocytosis as an independent risk factor for DFS (p = 0.1355). In our retrospective study, elevated platelet counts at time of diagnosis were associated with poor prognosis in breast cancer. We hypothesize that platelets may contribute to the pathophysiology of hematogenous metastasis. PMID- 12783125 TI - Thrombosis in children. PMID- 12783126 TI - Rapid determination of apolipoprotein E genotypes using two-sided allele-specific primer pairs. PMID- 12783127 TI - A caution on the use of murine hemophilia models for comparative immunogenicity studies of FVIII products with different protein compositions. PMID- 12783129 TI - Long-term survival of Taiwanese patients with hepatocellular carcinoma after combination therapy with transcatheter arterial chemoembolization and percutaneous ethanol injection. AB - BACKGROUND AND PURPOSE: Several studies have shown a superior effect of combination therapy with transcatheter arterial chemoembolization (TACE) and percutaneous ethanol injection (PEI) compared with either monotherapy for the treatment of advanced hepatocellular carcinoma (HCC), but there have been no reports on combination treatment from Taiwan. This study investigated the long term survival and prognostic factors of HCC patients treated with TACE/PEI combination therapy. METHODS: A total of 153 consecutive HCC patients, with tumor sizes between 2 and 3 cm in 47 patients, between 3 and 5 cm in 66 patients, and between 5 and 13 cm in 40 patients, who received TACE/PEI combination therapy were included in this retrospective study. The mean follow-up duration was 23 +/- 17 months (range, 1 to 78 months). RESULTS: The 1-, 2-, 3-, 4-, 5-, and 6-year cumulative survival rates for the patients were 78%, 54%, 40%, 22%, 12%, and 5%, respectively. Multivariate analysis using Cox's proportional hazards model showed that the stage of cirrhosis (Child's class B or C vs class A) was the only factor that significantly affected the survival rate (p = 0.02) [relative risk, 2.10; 95% confidence interval, 1.12 to 3.96]. Univariate analysis showed that survival was poorer in patients with tumors greater than 5 cm than in patients with tumors 2 to 5 cm in largest dimension; this difference was not significant in the multivariate analysis. No serious complications were observed during or after treatment. CONCLUSIONS: TACE combined with PEI is an alternative treatment for patients with larger HCC who are not suitable for surgical resection. A superior outcome can be expected in patients with Child's class A cirrhosis. PMID- 12783130 TI - Association between levels of TNF-alpha and TNF-alpha promoter -308 A/A polymorphism in children with Kawasaki disease. AB - BACKGROUND AND PURPOSE: Tumor necrosis factor-alpha (TNF-alpha) has been shown to play a central role in the pathogenesis of vasculitis in Kawasaki disease (KD). We investigated the serum levels of TNF-alpha and soluble TNF receptor 1 (STNFR1) levels, and genetic polymorphisms of the TNF-alpha promoter gene in children with KD to delineate the genetic basis of KD. METHODS: A total of 18 children (12 boys and 6 girls) with KD were studied, 9 of whom had the complication of coronary artery lesion (CAL) within 30 days after the onset of symptoms. Serum levels of TNF-alpha and STNFR1 were assayed by enzyme-linked immnosorbent assay, and DNA polymorphisms of the 5' flanking region of TNF-alpha promoter gene at position 308 [guanine (G) to adenine (A)] and -238 (G to A) were studied by direct nucleotide sequencing. RESULTS: The serum TNF-alpha level in KD patients was 113 +/- 209.9 pg/mL (range, 2.0 to 756.9 pg/mL; median, 24.7 pg/mL; normal, < 10 pg/mL). The serum levels of STNFR1 in KD (4255 +/- 2425 pg/mL) were higher than those of the control group (160 +/- 116 pg/mL). Allele frequencies of -308A and 238A were 11.1% and 0% in the KD patients, and 0% and 3.1% in the control group. Neither TNF-alpha promoter polymorphism nor any significant risk factor for CAL was identified in KD patients. One patient, who was homozygous for -308A, showed the highest TNF-alpha level and elevated STNFR1 level but had no evidence of CAL. Positive correlations were found between serum levels of STNFR1 and C-reactive protein (r = 0.731, p = 0.007), and between STNFR1 and leukocyte counts at admission (r = 0.620, p = 0.008). CONCLUSIONS: Increased serum levels of TNF alpha and STNFR1 were found in KD patients but there was no correlation between these levels. The relationship between the pathogenesis of KD and TNF-alpha gene promoter -308G to A mutation towards cytokine production remains to be clarified. PMID- 12783131 TI - Vitamin C protects against lysophosphatidylcholine-induced expression of monocyte chemoattractant protein-1 in cultured human umbilical vein endothelial cells. AB - BACKGROUND AND PURPOSE: It is well documented that oxidized low-density lipoproteins (LDLs) can stimulate human vascular endothelial cells to produce monocyte chemoattractant protein-1 (MCP-1). Vitamin C is known to be an important antioxidant for vasodilatation. The purpose of this study was to determine whether pretreatment with vitamin C could protect against oxidized-LDL-induced expression of MCP-1 in cultured human umbilical vein endothelial cells (HUVECs). METHODS: Cultured HUVECs were used for desired experiments before passage 4. Lysophosphatidylcholine (lysoPC), an oxidized component of LDL, was designated as the stimulator for MCP-1 synthesis from cultured HUVECs. MCP-1 concentrations in the cultured media were determined by enzyme-linked immunosorbent assay. MCP-1 RNA was evaluated by a semi-quantitative reverse transcriptase-polymerase chain reaction. RESULTS: HUVECs secreted MCP-1 within 30 minutes after exposure to 50 microM lysoPC. Compared with samples treated with lysoPC alone, pretreatment with vitamin C in concentrations of 50, 100, 150, and 200 microM, reduced levels of MCP-1 in the culture medium by 44%, 51%, 60%, and 67%, respectively, while levels of MCP-1 mRNA decreased by 15%, 18%, 80%, and 82%, respectively. CONCLUSIONS: Our findings imply that pretreatment with vitamin C can suppress lysoPC-induced expression and secretion of MCP-1 in cultured HUVECs. Therefore, vitamin C is protective against lysoPC-mediated inflammatory insults to the vascular endothelium in vitro. PMID- 12783132 TI - Left ventricular dysfunction is associated with CD4 lymphocyte count rather than opportunistic infection in human immunodeficiency virus infection. AB - BACKGROUND AND PURPOSE: Left ventricular (LV) dysfunction is often found in the early stage of human immunodeficiency virus (HIV) infection and deteriorates with disease progression. CD4 lymphocyte count and opportunistic infection are the major indicators for the clinical staging of HIV infection. This study investigated the association of these indicators with LV dysfunction in the clinical course of HIV infection. METHODS: HIV-positive patients without cardiac manifestations consecutively admitted from May 1998 to April 1999 were enrolled in the study. Echocardiographic LV function evaluation and measurement of CD4 lymphocyte count were performed. Parameters for LV systolic and diastolic functions were compared between patients with CD4 lymphocyte count >or= 200/microL and those with CD4 < 200/microL. In patients with CD4 < 200/microL, LV function was further correlated with the presence or absence of opportunistic infections. RESULTS: Ninety eight HIV-positive patients including 52 with CD4 >or= 200/microL and 46 with CD4 < 200/microL were studied. One half of the 46 patients with CD4 < 200/microL had active opportunistic infections. We found that LV fractional shortening, ejection fraction, and isovolumic relaxation time were all significantly lower in the patients with CD4 < 200/microL compared with those with CD4 >or= 200/microL. Moreover, these LV systolic and diastolic dysfunctions were positively correlated with decreased CD4 lymphocyte count. In contrast, no difference was found in these parameters between patients with and without opportunistic infections. In multiple regression analysis, CD4 lymphocyte count was found to be the only factor for predicting the LV systolic and diastolic dysfunction. CONCLUSIONS: Both LV systolic and diastolic function deteriorate as the CD4 lymphocyte count decreases in HIV infection. Opportunistic infection seems to have a limited role in the pathogenesis of LV dysfunction in advanced HIV infection. PMID- 12783133 TI - Clinical characteristics of patients with thyrotropin-secreting pituitary adenoma. AB - BACKGROUND AND PURPOSE: Thyroid-stimulating hormone (thyrotropin, TSH)-secreting pituitary adenoma is a very rare cause of hyperthyroidism. Diagnosis of this condition is often delayed due to lack of availability of TSH radioimmunoassay (RIA), the failure to recognize the utility of RIA and the incorrect attribution of the condition to other causes of thyrotoxicosis. This retrospective study analyzed the clinical characteristics of patients with this disorder treated from 1991 to 2002. METHODS: Seven patients (6 females, 1 male; mean age, 48 years; range, 33 to 72 years) with a diagnosis of TSHsecreting pituitary adenoma based on detectable TSH levels with high serum free thyroid hormone or triiodothyronine concentrations and pituitary lesions found on neuroimaging were included in this study. Patient records including clinical features, endocrine studies, immunohistochemistry studies, and response to treatment were reviewed. RESULTS: All 7 patients had hyperthyroidism, elevated free thyroxine or triiodothyronine levels, and unsuppressed levels of TSH. Imaging studies demonstrated a pituitary mass or lesion in all patients. Six patients had macroadenomas and 1 patient had a microadenoma. One of the patients had coexisting acromegalic features and hypersecretion of growth hormone was diagnosed. All of the patients had been treated with thionamides or thyroidectomy for presumed primary hyperthyroidism. Serum alpha-subunit level was uncharacteristically normal in 2 patients and elevated in 1 patient. Alpha-subunit/TSH molar ratios were elevated in 3 patients. Five patients underwent transsphenoidal adenomectomy but only one of them remained well-controlled at follow-up. Three patients received administration of somatostatin analogs and they achieved normalization of serum TSH and free thyroid hormones during the period of therapy. CONCLUSIONS: TSH immunoassay has an important role in the evaluation of hyperthyroid patients to determine the presence of inappropriate secretion. TSH-secreting pituitary adenoma exhibits heterogeneity in clinical presentation, hormonal expression and therapeutic response. PMID- 12783134 TI - MR cholangiopancreatography: prospective comparison of 3-dimensional turbo spin echo and single-shot turbo spin echo with ERCP. AB - BACKGROUND AND PURPOSE: Magnetic resonance cholangiopancreatography (MRCP) is a non-invasive technique for examination of the biliopancreatic tract. Respiratory triggered 3-dimensional turbo spin echo (3DTSE RT) and breath-hold thick slab single-shot turbo spin echo (ssTSE BH) are both useful MRCP techniques. The purpose of this study was to compare these 2 sequences with endoscopic retrograde cholangiopancreatography (ERCP) in patients with biliary tract disease. METHODS: Forty four patients with suspected biliary obstruction were recruited to receive MRCP within 3 days before ERCP. MRCP was performed using both 3DTSE RT with maximum intensity projection images and ssTSE BH. ERCP was performed and assessed by 2 endoscopists. RESULTS: MRCP was successfully performed in all patients, whereas ERCP failed in 6 patients (13.6%). MRCP was effective in detecting the presence of choledocholithiasis in 13 of 14 patients, ERCP in 12 of 12, and 2 failed ERCP. MRCP was effective in detecting benign biliary obstruction in 18 of 19 patients, and ERCP in 15 of 15, but 4 patients failed ERCP and choledocholithiasis was misdiagnosed by MRCP in 1 patient. Both MRCP and ERCP correctly diagnosed malignant bile duct obstruction in 10 of 11 patients, and both misdiagnosed that condition as benign obstruction in 1 patient. There was no significant difference between MRCP and successful ERCP in detecting lesions. MRCP was significantly better than ERCP when both successful and failed ERCP were encountered (p = 0.0498). Both 3DTSE RT and ssTSE BH produced the same results in depicting the biliary ducts and lesions in 37 patients (84.1%). Four patients (9.1%) showed better images on 3DTSE RT, whereas 3 patients (6.8%) showed better images on ssTSE BH. CONCLUSIONS: 3DTSE RT and the ssTSE BH were complementary to each other in MRCP studies. Using these 2 techniques, MRCP has a high successful rate and diagnostic accuracy when compared with ERCP in detecting bile duct disease. PMID- 12783135 TI - Pit pattern analysis by magnifying chromoendoscopy for the diagnosis of colorectal polyps. AB - BACKGROUND AND PURPOSE: The development of magnifying chromoendoscopy has facilitated the observation of mucosal pit patterns. This study investigated the value of this technology in predicting the histologic findings of colorectal lesions. METHODS: A total of 954 colorectal polyps were included. After identifying the lesions at colonoscopy, 0.2% indigocarmine solution was sprayed and then the zoom apparatus was switched to make a magnified view of the stained crypt orifice at a maximum 100 times magnification. The observed pit patterns were classified into 6 categories (I, II, IIIL, IIIS, IV, and V) according to Kudo's classification. Type I and II were designated as non-neoplastic patterns whereas other types were neoplastic. Correlation of the pit pattern with the findings of histologic examinations of resected or biopsied polyps was performed. RESULTS: There were 678 diminutive ( or =1.7 mmol/l), low levels of HDL-cholesterol (<0.9 mmol/l) and abdominal obesity (waist/hip ratio>0.9). The risk of low HDL-cholesterol was doubled in shift workers, (odds ratio (OR): 2.02, 95% confidence interval (95% CI): 1.24-3.28) after being adjusted for age, socio-economic factors, physical activity, current smoking, social support and job strain. High levels of triglycerides were also significantly associated with shift work (OR: 1.40, 95% CI: 1.08-1.83). The OR for abdominal obesity was 1.19, (95% CI: 0.92-1.56). The prevalence of hyperglycaemia (serum glucose > or =7.0 mmol/l) was similar in day and shift workers. No significant interaction was seen between shift work and abdominal obesity with regard to the associations with triglycerides and HDL-cholesterol. CONCLUSIONS: We found a significant association between shift work and lipid disturbances (i.e. low HDL-cholesterol and high triglyceride levels). We did not find any association with hyperglycaemia. PMID- 12783236 TI - Federal government regulation of occupational skin exposure in the USA. AB - There are at least 14 federal regulations and three agencies that are involved in the regulation of occupational skin exposures in the USA. The Environmental Protection Agency (EPA) requires the reporting of health effects information on chemicals, and such information is used to assess the risks of human and environmental exposure. The health effects information and any resulting risk assessments are generally available to the public. A fair amount of this information relates to skin irritation, sensitization, and dermal absorption. The EPA can require the submission of new data necessary for it to carry out its risk assessments, and has the authority to ban hazardous chemicals for certain uses. The Food and Drug Administration (FDA) regulates the correct labeling of cosmetics and requires safety and efficacy data on new products that are claimed to have preventive or health benefits. Commercial distribution of topical skin care and protection products, therefore, can be potentially scrutinized by the FDA, which can control the use of hazardous chemicals in such products. The Occupational Safety and Health Administration (OSHA) has the most direct contact with workplaces through its field inspection compliance activity, which is directed at the reduction of workplace injuries and illnesses. Our analysis suggests that although considerable amounts of health effects information is generated and available, such information may not always be adequately conveyed to the end users of chemical products. In addition, the most effective and practical means of preventing exposure is often not apparent or generally known. Current regulations may have created a reliance on use of chemical protective equipment that may not always be the best approach to protecting workers. Lack of performance criteria that are measurable has hampered industry from objectively assessing skin exposures. This lack of performance criteria or guidance has also hindered the implementation of prevention strategies and a critical assessment of their effectiveness. Better guidance from regulatory agencies directed at performance-based control of occupational skin hazards is presently needed. PMID- 12783237 TI - Workers' assessments of manual lifting tasks: cognitive strategies and validation with respect to objective indices and musculoskeletal symptoms. AB - OBJECTIVES: To determine the different cognitive strategies adopted by workers in assessing the effects of lifting-task parameters on effort, and to validate workers' assessments. METHODS: Questionnaires were administered to 217 male workers with varied levels of experience in manual handling. Workers were asked to assess the effects of lifting on perceived effort, using linguistic descriptors (e.g., light, heavy), and to determine the physical meaning of such descriptors. In addition, each worker assessed on-the-job effort, perceived risk of injury and work dissatisfaction, and musculoskeletal outcomes in a cross sectional design. RESULTS: Perceived physical effort was significantly associated with lifting variables. Results indicated that the three-cluster strategy is the best performer. Weight of load emerged as the most influential factor that impacted on effort in the most dominant cluster (close to 50% of the observations). The second cluster (25% of the observations) demonstrated that weight, horizontal distance, and twisting angle, contributed equally to effort, and the third cluster had weight and vertical travel distance as the most important variables (with travel distance being more important). Perceived effort was significantly associated with objective indices (i.e., biomechanical lifting equivalent and NIOSH lifting index), and musculoskeletal symptoms in eight body parts. CONCLUSIONS: Cognitive reasoning of experienced workers may be used as an active device for the evaluation of strenuous physical activities such as lifting tasks. Lifting activities are significantly associated with musculoskeletal symptoms, not only in the lower-back region, but also in seven other body parts; and effort may integrate the effects of both physical (lifting tasks) and non physical (i.e., work dissatisfaction) factors, as well as perception of risk. PMID- 12783239 TI - A very long ACTBP2 (SE33) allele. AB - Analysing a buccal swab we found a long allele in the STR system ACTBP2. For confirmation we sequenced the isolated PCR product and found a sequence structure common in alleles of type III. Based on the repeat array the new allele is assigned as allele "49". PMID- 12783238 TI - First case of feline spongiform encephalopathy in a captive cheetah born in France: PrP(sc) analysis in various tissues revealed unexpected targeting of kidney and adrenal gland. AB - Feline spongiform encephalopathy (FSE), affecting domestic and captive feline species, is a prion disease considered to be related to bovine spongiform encephalopathy. Here we report an immunohistological analysis of the first FSE affected cheetah born in France. The duration of clinical signs, of which ataxia was the main one, was about 8 weeks. The distribution of abnormal prion protein (PrP(sc)) was studied by immunohistochemistry within 27 different tissues. Different antibodies were used to visualise abnormal PrP deposits in situ. PrP(sc )accumulation was detected in the central nervous system (cerebral cortex, cerebellum, brain stem, spinal cord, retina), in peripheral nerves and in lymphoid organs. PrP(sc) deposits were not observed within the enteric nervous system nor in several other organs, such as pancreas, ovary, liver and muscle. More interestingly, unusual PrP(sc )deposits were observed within the zona fasciculata/reticularis of the adrenal gland and within some glomeruli of the kidney raising the question of possible PrP(sc) excretion. The sympathetic innervation of these two organs was visualised and compared to the distribution of PrP(sc) deposits. Our results suggest the possibility that the infectious agent is spread by both haematogenous and nervous pathways. PMID- 12783242 TI - Magnetic resonance imaging after reduction for congenital dislocation of the hip. AB - BACKGROUND: Reliable concentric reduction of the femoral head and subsequent retention in a centred position are indispensable preconditions for the remodelling of the acetabulum in developmental dysplasia of the hip (DDH) and to prevent damage to the hip joint, i.e. avascular necrosis. The objective of this study is to evaluate the necessity of verifying the reduced position of the articulation in the plaster cast. METHOD: MRI was carried out in 15 infants with 21 unstable hip joints after reduction under arthrographic control and fixation in a plaster cast in the 'human' position with the hips flexed above 90 degrees and abducted to 50 degrees or 60 degrees. When the reduction was found to be inadequate-the hip still partially or completely dislocated-the plaster cast was removed, reduction repeated, a new cast applied, and MRI carried out again. RESULTS: After primary reduction, 1 of 21 hips was dislocated, and 2 showed unsatisfactory reduction. Three hip joints out of 21 (14.3%) were not fixed in the plaster cast in the optimal centred position. CONCLUSION: In view of the number of inadequate reductions in plaster casts, we recommend verifying the position of the hip joint by MRI. This MRI documentation should be established as a standard examination post-reduction. PMID- 12783241 TI - Pre- and intraoperative assessment of mid-cord erythroleukoplakias: a prospective study on 52 patients. AB - A pre- and intraoperative evaluation by videolaryngostroboscopy (VLS) and saline infusion (SI) into Reinke's space was prospectively applied to a cohort of 52 patients with mid-vocal cord erythroleukoplakias observed at our institution between January 1997 and June 1999. These tests were done to predict the invasion of the layered structure of the lamina propria and consequently determine the deep extent of the excisional biopsy to be carried out. According to the results of these examinations, carbon dioxide laser resections were performed as follows: 25 Type I, 17 Type II and 10 Type III cordectomies according to the European Laryngological Society Classification. Histopathologic diagnoses were keratosis without atypia in 13 patients, keratosis with mild, moderate or severe dysplasia in 8, 3 and 5, respectively, microinvasive carcinoma in 12 and invasive carcinoma in 11. By comparing the type of resection with the pathologic diagnosis obtained on the specimen and its surgical margins, we estimated the number of under- and overtreatments. In addition, an assessment was made on specificity, sensitivity, positive and negative predictive values and accuracy of VLS and SI, both independently and in combination. Thirty-eight patients (73%) received the correct type of resection, 1 patient (2%) an undertreatment and 13 (25%) an overtreatment. VLS and SI showed a specificity, sensitivity, positive and negative predictive values and accuracy of 82, 100, 94, 100, 97% and 90, 83, 86, 87 and 87%, respectively. The VLS/SI combination raised the values to 89, 100, 88, 100 and 94%. The 4-year overall and disease-free survival and ultimate local control with laser alone were 90, 94 and 100%. PMID- 12783243 TI - Transscaphoid-transtriquetral perilunate fracture dislocation: report of a case and review of the literature. AB - INTRODUCTION: A transscaphoid and transtriquetral perilunate fracture dislocation is fairly rare among the known cases of perilunate fracture dislocations, and the details of the initial treatment and outcome of this injury have never been reported. MATERIALS AND METHODS: A 21-year-old, right-handed man presented with fractures at the proximal third of the scaphoid and at the mid-body of the triquetrum with an associated dorsal perilunate dislocation after a fall onto his outstretched hand. Under general anesthesia, closed reduction was attempted with 3 kg of traction applied by means of finger traps. After anatomical reduction was achieved, percutaneous fixation was applied to both the triquetrum and scaphoid using cannulated screws. A short arm thumb spica splint was applied for 2 weeks, and part-time splinting was continued for an additional 3 weeks. The patient subsequently underwent 3 months of intensive range-of-motion and muscle strengthening exercises. RESULTS: At the final follow-up examination 68 months after the initial operation, the arc of motion of the right wrist, 150 degrees (extension plus flexion arc), and grip strength, 41 kg, were 94% and 103% of the values for the unaffected wrist, respectively. Radiographs showed a bony union of the scaphoid and triquetrum, and no sign of avascular necrosis in the proximal scaphoid fragment, as well as other carpi. No midcarpal or radiocarpal degenerative arthritis was observed, and the normal carpal bone relationships were still maintained, with a scapholunate angle of 48 degrees and a scapholunate gap of 2 mm. CONCLUSION: We recommend closed reduction and percutaneous screw fixation of the scaphoid, as well as the triquetrum in this case, to minimize the interruption of the blood supply to the carpus and also to obtain rigid fixation during the procedure. PMID- 12783245 TI - Limb salvage after subtotal supramalleolar amputation by initial shortening followed by tibial lengthening. AB - BACKGROUND: We present a patient with a subtotal traumatic supramalleolar amputation of the leg, which was initially treated by a vascular reconstruction with deliberate bone and soft-tissue shortening. METHODS: To correct the ensuing complex deformity, which consisted of a varus hindfoot, leg length discrepancy and equinus, a staged reconstruction was planned. Initially, the hindfoot varus, in presence of a stiff ankle, was corrected by a supramalleolar osteotomy, followed by a Wagner distraction and finally a correction of the equinus. RESULTS: After a relatively long period of normal functioning, she regained painful minimal ankle function, which necessitated ankle fusion and correction of a pronation deformity. At the most recent follow-up 13 years after the injury, the patient is fully functional and has near normal leg length. CONCLUSION: Although a mangled lower extremity is often a candidate for primary amputation allowing early rehabilitation, in certain cases a good result can be obtained by a creative strategy. PMID- 12783246 TI - Glial fibrillary tangles in diffuse neurofibrillary tangles with calcification. AB - In the present study the occurrence and distribution of glial fibrillary tangles (GFT) and their related structures in diffuse neurofibrillary tangles with calcification (DNTC) were investigated using Gallyas-Braak (GB) stain. Six cases neuropathologically diagnosed as DNTC were studied (two males and four females). The age at death ranged from 56 to 73 years, with an average of 63.5+/-7.5 years. GFT were classified morphologically, and their immunoreactivites for tau and ubiquitin were examined. Glial cells with GFT were identified with astrocytes and oligodendrocytes by immunostain for glial fibrillary acidic protein and transferrin, respectively. A small number of coiled bodies detected within the oligodendrocytes in the white matter of the cerebrum were positive for tau and ubiquitin. Cell clusters of thorn-shaped astrocytes were detected in the subcortical and subpial regions where gliosis occurred. Thorn-shaped astrocytes were positive for tau, but negative for ubiquitin. A small number of tuft-shaped astrocytes detected prominently in the temporal cortex and amygdala with numerous neurofibrillary tangles were positive for tau and ubiquitin. All three types of GFT were detected, especially in the temporal and limbic lobes, which were the most severely affected sites in DNTC. Moreover, various-shaped neurofibrillary tangles, aggregated rods and some argyrophilic threads were differentiated from GFT. They were positive for GB, but not detected within the glial cells. PMID- 12783247 TI - Clinical and neuropathological correlates of Lewy body disease. PMID- 12783249 TI - Expression of alpha-, beta-, and gamma-synuclein in glial tumors and medulloblastomas. AB - alpha-, beta- and gamma-synuclein are highly homologous proteins that are found predominantly in neurons. Abnormal accumulation of synucleins has been associated with diseases of the central nervous system particularly Parkinson's disease. Immunoreactivity of alpha-synuclein is demonstrated in brain tumors with neuronal differentiation and in schwannomas, whereas gamma-synuclein has been demonstrated in breast and ovarian carcinomas. The immunoreactivity of synucleins has not been described in glial tumors. Immunoreactivity of synucleins in glial cells in culture and in pathological conditions, however, suggests that synucleins may be expressed by glial tumors. We studied the expression of alpha-, beta-, and gamma synuclein in 84 human brain tumors (24 ependymomas, 31 astrocytomas, 8 oligodendrogliomas, and 21 medulloblastomas) by immunohistochemistry. Our study demonstrates immunoreactivity for gamma-synuclein in high-grade glial tumors; immunoreactivity is found in all anaplastic ependymomas but in only 33% of ependymomas and 16% of myxopapillary ependymomas. Immunoreactivity for gamma synuclein is noted in 63% of glioblastomas but not in other astrocytic tumors. Of medulloblastomas, 76% have immunoreactivity for either alpha- or beta-synuclein or both; no immunoreactivity for gamma-synuclein is seen in medulloblastomas. PMID- 12783250 TI - A novel leukoencephalopathy associated with tau deposits primarily in white matter glia. AB - A 79-year-old woman had a 10-year history of dementia, initially presenting as non-fluent aphasia. Magnetic resonance imaging showed frontal atrophy (left greater than right) and hyperintense foci within white matter. Neuropathologically, there was severe frontal atrophy due to cortical neuronal loss with spongy change and to an even greater loss of white matter that contained prominent eosinophilic deposits. The deposits were immunoreactive for phosphorylated tau, non-reactive for Abeta and alpha-synuclein and equivocally or weakly reactive for ubiquitin. They stained with the Gallyas, Bielschowsky, and Bodian techniques. Ultrastructural examination revealed the deposits to be composed of straight filaments with a diameter of approximately 10 nm, primarily in white matter glia. Moderate loss of neurons in substantia nigra and numerous argyrophilic threads in gray and particularly white matter were noted. The precise relationship between this disorder and other frontotemporal degenerations/tauopathies, as well as the pathogenetic basis of the leukoencephalopathy, remains to be determined. PMID- 12783252 TI - Prognostic relevant factors within pT3 rectal carcinomas: depth of invasion or circumferrential resection margins? PMID- 12783251 TI - Prognostic significance of depth of gross or microscopic perirectal fat invasion in T3 N0 M0 rectal cancers following sharp mesorectal excision and no adjuvant therapy. AB - BACKGROUND AND AIMS: Although the standard of care for T3 and/or N1-2 rectal cancers includes adjuvant chemoradiation, it is unclear whether T3 N0 patients with limited microscopic perirectal fat invasion warrant further therapy. Our aim was to determine the prognostic significance of gross perirectal fat invasion, or depth of microscopic perirectal fat invasion, in T3 N0 rectal cancers following sharp mesorectal excision and no adjuvant therapy. PATIENTS AND METHODS: Utilizing a prospective database, the medical records of 108 patients who underwent a potentially curative resection for T3 N0 rectal cancer between June 1986 and December 1994 were analyzed. All pathological specimens were re-reviewed by a single pathologist, and extent of perirectal fat invasion was measured in millimeters. Patients who received either preoperative or postoperative adjuvant therapy were excluded. RESULTS: Macroscopic perirectal fat invasion (T3 gross) was present in 49 cases, absent in 40 cases (T3 microscopic), and not reported in 19 cases. Rectal cancers were stratified by extent of measured perirectal fat invasion into 3 mm or less and more than 3 mm. Five-year overall and local recurrence rates for the entire group were 19% and 8%, respectively. The disease free survival, disease-specific survival, and overall recurrence for rectal cancers with 3 mm or less invasion vs. more than 3 mm invasion, or T3 gross vs. T3 microscopic, were not statistically different. CONCLUSION: Our data suggest that the extent of gross, or microscopic perirectal fat invasion (defined as >3 or 2.7 mmol/l] were treated with a single infusion of ibandronate (2 or 4 mg) or pamidronate (15, 30, 60, or 90 mg) on day 0. The dose was dependent on the severity of hypercalcemia (baseline CSC level). CSC was assessed daily until day 4, then at intervals until day 28. The primary endpoint was lowering of CSC at day 4. Secondary endpoints included the number of patients responding and time to re-increase following response. MAIN RESULTS: Using the CSC baseline approach, the most frequently administered doses were 4 mg ibandronate (78.4%) and 60 mg pamidronate (50.0%). Mean lowering of CSC at day 4 was 0.6 mmol/l for ibandronate and 0.41 mmol/l for pamidronate. The 95% confidence interval for the difference ibandronate pamidronate had a lower limit of 0.05 mmol/l, indicating that ibandronate was as effective as pamidronate. The number of patients responding to the two agents was also similar; 76.5% of ibandronate patients and 75.8% of pamidronate patients were rated as responders after the first dose of study medication. The median time to re-increase after response was longer for ibandronate (14 days) than pamidronate (4 days) ( P=0.0303). In the subgroup of 17 patients with high baseline CSC (>3.5 mmol/l), ibandronate appeared to be more effective than pamidronate. The safety profile of both agents was similar. CONCLUSIONS: Ibandronate is at least as effective as pamidronate in the treatment of HCM. Furthermore, in patients with higher baseline CSC ibandronate appears to be more effective than pamidronate. The duration of response is significantly longer with ibandronate than pamidronate. PMID- 12783290 TI - Acute respiratory distress syndrome after chemotherapy for lung metastases from non-seminomatous germ-cell tumors. AB - GOALS: To describe an acute respiratory distress syndrome (ARDS) occurring after chemotherapy for non-seminomatous germ-cell tumors (NSGCT) with diffuse lung metastases, we conducted a retrospective study in a 15-bed intensive care unit (ICU) in a comprehensive cancer center. PATIENTS AND METHODS: During a 10-year period, 16 consecutive patients with diffuse lung metastases from a NSGCT were admitted to the ICU for respiratory distress and high-risk chemotherapy. MAIN RESULTS: Nine patients developed acute respiratory failure requiring mechanical ventilation (MV) within 3 days of the initiation of chemotherapy, while the respiratory status of the seven other patients improved. The evolution was independent of tumor marker levels and the type of chemotherapy regimen. The SAPS II score did not accurately describe the severity of this population. The only predictor of intubation was the initial PaO2/FiO2 ratio upon admission to the ICU. Six out of seven patients who did not require MV were discharged alive from the hospital, whereas all but one patient requiring MV died. Refractory hypoxemia and ventilator-associated pneumonia were the leading causes of death. CONCLUSIONS: Acute respiratory distress in patients with lung metastases from NSGCT is a rare cause of ARDS. Chemotherapy could be responsible for triggering the respiratory worsening. Patients with severe respiratory insufficiency (PaO2 <70 mmHg on room air) on admission to hospital should be promptly transferred to the ICU for the first chemotherapy course. PMID- 12783291 TI - Assessing temperature and humidity conditions for dairy cattle in Cordoba, Argentina. AB - Temperature and humidity conditions affect livestock production in Central Argentina. This study evaluates the risk of thermal stress affecting dairy production. The temperature-humidity index (THI) was used to analyze the regional and seasonal effects of temperature and humidity. Statistically, the THI was found to be normally distributed. The probability of occurrence of a daily THI higher than 72 was 40% for Rio Cuarto during January. Regional variability of THI indicates a low risk of harmful extreme thermal stress conditions. The probability of THI being 78 or above ranges between 4% and 10% for the main dairy region of Cordoba during January. Also, in January and February, dairy production losses between 3 and 4 l cow(-1) day(-1) could be expected with a frequency of 5% in Rio Cuarto and 15% in Villa de Maria de Rio Seco. PMID- 12783292 TI - Two statistical approaches to forecasting the start and duration of the pollen season of Ambrosia in the area of Lyon (France). AB - The aim of the present study was to forecast the start and duration of the pollen season of Ambrosia from meteorological data, in order to provide early information to allergists and allergic people. We used the airborne pollen data from Lyon (France), sampled using a Hirst trap from 1987 to 1999, and the meteorological data for the same period: air temperature (minimal, maximal, and average), rainfall, relative humidity, sunshine duration and soil temperature. Two forecasting models were used, one summing the temperatures and the other making use of a multiple regression on 10-day or monthly meteorological parameters. The start of the pollen season was predicted with both methods, results being more accurate with the regression (the errors between the predicted and the observed SDP ranging from 0 to 3 days). The duration of the pollen season was predicted by a regression model, errors ranging from 0 to 7 days. The models were later tested with satisfactory results from 2 additional years (2000 and 2001). Such forecasting models are helpful for allergic people, who have to begin their anti-allergic treatment before the start of the pollen season and not when the symptoms have appeared, since a preventive treatment is more efficient than a curative one. The regression allows predictions to be made 3-5 weeks in advance and so it is of particular interest. The forecasts will be broadcast on the Internet. PMID- 12783293 TI - Renal phosphate handling of premature infants of 23-25 weeks gestational age. AB - Premature infants with low serum phosphate concentrations (<2 mmol/l) are at risk for osteopenia. Therefore, serum phosphate levels in premature infants should be kept above 2 mmol/l. Premature infants of 26-31 weeks gestational age (GA) have renal phosphate threshold concentrations (Tp/GFR) in the range of normal serum phosphate values (2 mmol/l). Therefore, these infants show significant urinary phosphate excretion only when serum phosphate levels are normal, and urinary phosphate excretion can be used to monitor phosphate supplementation. However, few data are available on extremely premature infants of 23-25 weeks GA. The objective of this study was to compare Tp/GFR levels in infants of 23-25 weeks GA to those in infants of 26-31 weeks GA. We retrospectively evaluated case notes of 12 infants of 23-25 weeks GA and compared them to 19 infants of 26-31 weeks GA. Tp/GFR was calculated from simultaneous measurements of urinary phosphate, urinary creatinine, serum phosphate, and serum creatinine. Tp/GFR values 3-5 weeks postnatally were lower in infants of 23-25 weeks GA (1.06+/-0.36 mmol/l, p<0.001) than in infants of 26-31 weeks GA (1.76+/-0.26 mmol/l). Near term (35-37 weeks postmenstrual age), there was no significant difference between Tp/GFR values in infants of 23-25 weeks GA (1.83+/-0.32 mmol/l) and in infants of 26-31 weeks GA (2.05+/-0.22 mmol/l). We conclude that at 3-5 weeks postnatally, infants of 23-25 weeks GA are at risk for low Tp/GFR values, leading to urinary phosphate excretion even in the presence of low serum phosphate levels. In these infants, serum phosphate levels should be monitored, and phosphate supplementation should be adjusted to keep serum phosphate levels above 2 mmol/l. PMID- 12783294 TI - Flowering phenology and compensation for herbivory in Ipomopsis aggregata. AB - The mechanisms and circumstances that affect a plant's ability to tolerate herbivory are subjects of ongoing interest and investigation. Phenological differences, and the timing of flowering with respect to pollinators and pre dispersal seed predators, may provide one mechanism underlying variable responses of plants to herbivore damage. The subalpine wildflower, Ipomopsis aggregata, grows across a wide range of elevations and, because phenology varies with elevation, phenological delays associated with elevation may affect the ability of I. aggregata to compensate for or tolerate browsing. Thus, we examined the response of I. aggregata to herbivory across an elevation gradient and addressed the interactions among phenological delays imposed by damage, elevation, pre dispersal seed predation and pollination, on I. aggregata's compensatory response. Among high and low elevation populations in areas near the Rocky Mountain Biological Laboratory (RMBL) in Gothic, Colorado, we compared the responses of naturally browsed, artificially browsed (clipped), and unbrowsed (control) plants of I. aggregata. We compared responses in the date of initiation of flowering, timing of peak bloom, floral display, nectar production and sugar concentration, oviposition and fruit destruction by the pre-dispersal seed predator Hylemya sp. (Anthomyiidae), fruit production, and aboveground biomass production. Clipping had the greatest effect on reproductive success and clipped plants at high elevation exhibited the lowest tolerance for herbivory. The effects of browsing appear to be mediated by flowering phenology, and both browsing and elevation delayed flowering phenology. Time needed for regrowth delays flowering, and thus affects the overlap with seed predators and pollinators. As a result of delayed flowering, naturally browsed and clipped plants incurred lower rates of seed predation. In the absence of seed predation, plants would exhibit a lower tolerance to herbivory since naturally and artificially browsed plants had fewer fruits destroyed by Hylemya larvae. We provide additional evidence that, for populations near the RMBL, clipping and natural browsing do not have the same effect on I. aggregata plants. This may be due to the selection of larger plants by herbivores. Although under some conditions plants may tolerate browsing, in areas where the growing season is short a phenological delay imposed by damage is likely to significantly reduce plant fitness. Identifying the mechanisms that allow plants to tolerate herbivore damage will help to develop a general framework for understanding the role of tolerance in plant population and community dynamics, as well as plant-herbivore interactions. PMID- 12783295 TI - Reduced immunocompetence and cost of reproduction in common eiders. AB - Immunocompetence may be especially important in long-lived species where infectious organisms may have detrimental effects upon future reproductive value of hosts. The resource demand for immunocompetence may reduce resource availability for reproduction and a trade-off between these traits has therefore been proposed. Capital breeders, such as the common eider (Somateria mollissima), rely upon accumulated body reserves during reproduction. Eiders lose more than 40% of pre-breeding body mass during egglaying and incubation and many females abandon their ducklings to other females after hatching. Results from our observational study show that levels of leukocytes (i.e., lymphocytes, heterophils and heterophil/lymphocyte ratio) are not related to body mass early in the incubation period. However, eider females with low initial body mass showed signs of immunosuppression (i.e., decreased late levels of lymphocytes) and increased response towards stressors (i.e., increased heterophil/lymphocyte ratio) later in the incubation period. Moreover, females with low lymphocyte levels more frequently abandoned their brood, and females abandoning young had an increased return rate to the next breeding season. However, among brood abandoning females return rate was lower for the females with low lymphocyte levels. These results suggest that immunosuppression, as indicated by low lymphocyte levels, is a reproductive cost that may be partly compensated for by abandoning young. PMID- 12783296 TI - Ecological constraints on extra-pair paternity in the bluethroat. AB - The mating system of the bluethroat (Luscinia s. svecica) involves a high level of sperm competition, and consequently a high frequency of extra-pair paternity (EPP). There is considerable variation in the frequency of EPP. Over the course of ten study years in a population in Norway, the frequency has fluctuated between 7% and 33% of young, and 8% and 76% of broods. In this paper, we address the issue of whether ecological factors can explain some of the variation in EPP between years and broods. Factors include breeding density, breeding synchrony and two meteorological variables (ambient temperature and precipitation) during the period of peak female fertility. There were no significant relationships between annual averages of the four variables and the annual level of EPP, but the statistical power of these tests was low, due to a restricted sample (n=10 years). Focussing on individual broods, none of the factors had significant effects when including all broods in the analyses (including those with zero EPP). When limiting the analyses to broods with one or more extra-pair offspring, morning temperature had a significant effect on the frequency of EPP. When the temperature was low during the peak of the fertile period, EPP occurred less frequently. Male extra-pair behaviour was not generally constrained by synchrony between the fertile periods of the social mate and the extra-pair mate, but for a subgroup of males ("unattractive males") there appeared to be a trade-off between mate guarding and pursuing extra-pair copulations. Our results indicate that ecological conditions have no influence on whether or not a brood will contain extra-pair offspring, but when EPP does occur, its frequency is influenced by air temperature. Possibly, there is a causal relationship between weather conditions and the frequency of extra-pair encounters in this species. The breeding season is initiated at a phenologically early stage, when large parts of the territories are still covered with snow, and cold weather conditions may force the birds to devote most of their time to maintenance and thereby constrain sexual activities. PMID- 12783297 TI - Immune responsiveness in adult blue tits: heritability and effects of nutritional status during ontogeny. AB - What is the relative contribution of genetic and various environmental factors to variation in the ability to mount an immune response? We measured antibody responsiveness to diphtheria-tetanus vaccine during the winter in free-ranging blue tits with a known nestling history to investigate (1) if nutritional status during the nestling stage has persistent effects on an individual's immune defence and (2) if immune responsiveness is heritable. There was no correlation between nutritional status during the nestling phase (measured as size-corrected body mass day 14 post-hatch) and antibody responsiveness as an adult. On the other hand, the heritability of responsiveness to diphtheria and tetanus, as estimated by parent-offspring regression, was 0.21+/-0.51 and 1.21+/-0.40 SE, respectively. Thus, while there was little evidence that natural variation in antibody responsiveness to these antigens reflected nutritional conditions during early life, responsiveness to at least one of the antigens (tetanus) had a strong genetic component. PMID- 12783298 TI - Density-dependent seed set in the Haleakala silversword: evidence for an Allee effect. AB - Plant species may be subject to Allee effects if individuals experience a reduction in pollination services when populations are small or sparse. I examined temporal variation in reproductive success of the monocarpic Haleakala silversword (Argyroxiphium sandwicense subsp. macrocephalum) over five years, to determine if plants flowering out of synchrony with most of the population (i.e., in low flowering years) exhibited lower percent seed set than synchronously flowering plants (i.e., those flowering in high flowering years). Through two pollination experiments conducted over multiple years, I also measured pollen limitation and self-incompatibility in this species. The number of flowering plants varied greatly among years, as did reproductive success. Percent seed set was significantly correlated with the number of plants flowering annually, such that plants flowering in high flowering years (1997 and 2001) exhibited significantly higher percent seed set than did plants flowering in low flowering years (1998-2000). In the 3-year pollen limitation study, plants flowering asynchronously were pollen-limited, whereas plants flowering synchronously were not. This species is strongly self-incompatible. Results of this study demonstrate that the Haleakala silversword experiences reduced reproductive success in low flowering years, and suggest that this Allee effect is pollinator mediated. Allee effects in plants are an understudied yet potentially important force with implications for the population dynamics and conservation of rare species. PMID- 12783299 TI - Temporal and spatial variations of gyne production in the ant Formica exsecta. AB - Social insects have become a general model for tests of sex allocation theory. However, despite tremendous interest in the topic, we still know remarkably little about the factors that cause dramatic differences in sex allocation among local populations. A number of studies have suggested that environmental factors may influence sex allocation in ant populations. In polygynous (multiple queens per nest) populations of the ant Formica exsecta, sex allocation is extremely male biased at the population level, with only a small proportion of nests producing any gynes (female reproductive brood). We analysed the proportion of gyne-producing nests in 12 F. exsecta populations during three successive breeding seasons and found considerable temporal and spatial variability in the proportion of gyne-producing nests. The populations differed in a number of characteristics, including elevation, nest density, size of the nest mound, and number of nests per population. However, the proportion of gyne-producing nests was not associated with any of these geographic and demographic variables. Moreover, differences between populations in the production of gynes were not consistent between years. Thus, the proportion of gyne-producing nests appears to vary stochastically, perhaps because of stochastic variations in environmental factors. For example, year-to-year variations in the proportion of gyne-producing nests were associated with differences in spring weather conditions between years. The finding that gyne production varies greatly between years suggests that it may not always be adaptive at a local scale. PMID- 12783300 TI - Markers informative for ancestry demonstrate consistent megabase-length linkage disequilibrium in the African American population. AB - Admixture mapping is a potentially powerful tool for mapping complex genetic diseases. For application of this method, admixed individuals must have genomes composed of large segments derived intact from each founding population. Such segments are thought to be present in African Americans (AA) and should be demonstrable by examination of linkage disequilibrium (LD). Previous studies using a variety of polymorphic markers have variably reported long-range LD or rapid decay of LD. To further define the extent and characteristics of LD caused by admixture in the AA population, the current study utilized a set of 52 diallelic markers that were selected for large standard variances between putative representatives of the founder populations. LD was examined in over 250 marker-pairs, including linked markers from four different chromosomal regions and an equal number of matched unlinked comparisons. In the representative founder populations, strong LD was not observed for markers separated by more than 10 kb. In contrast, results indicated significant LD ( P<0.001, D'>0.3) in AA over large genomic segments exceeding 10 centiMorgans (cM) and 15 megabases (Mb). Only marginally significant LD was present between unlinked markers in this population, suggesting that choosing appropriate levels of significance for admixture mapping can minimize false positive results. The ability to detect LD for extended chromosomal segments in AA decayed not only as a function of the distance between markers, but also as a function of the standard variance of the markers. This examination of several genomic segments provides strong evidence that appropriate selection of informative markers is a crucial prerequisite for the application of admixture mapping to the AA population. PMID- 12783301 TI - Increased prevalence of CFTR mutations and variants and decreased chloride secretion in primary sclerosing cholangitis. AB - Primary sclerosing cholangitis (PSC) and cystic fibrosis (CF) are both slowly progressive cholestatic liver diseases characterized by fibro-obliterative inflammation of the biliary tract. We hypothesized that dysfunction of the CF gene product, cystic fibrosis transmembrane conductance regulator (CFTR), may explain why a subset of patients with inflammatory bowel disease develop PSC. We prospectively evaluated CFTR genotype and phenotype in patients with PSC ( n=19) compared with patients with inflammatory bowel disease and no liver disease ( n=18), primary biliary cirrhosis ( n=17), CF ( n=81), and healthy controls ( n=51). Genetic analysis of the CFTR gene in PSC patients compared with disease controls (primary biliary cirrhosis and inflammatory bowel disease) demonstrated a significantly increased number of mutations/variants in the PSC group (37% vs 8.6% of disease controls, P=0.02). None of the PSC patients carried two mutations/variants. Of PSC patients, 89% carried the 1540G-variant-containing genotypes (resulting in decreased functional CFTR) compared with 57% of disease controls ( P=0.03). Only one of 19 PSC patients had neither a CFTR mutation nor the 1540G variant. CFTR chloride channel function assessed by nasal potential difference testing demonstrated a reduced median isoproterenol response of 14 mV in PSC patients compared with 19 mV in disease controls ( P=0.04) and 21 mV in healthy controls ( P=0.003). These data indicate that there is an increased prevalence of CFTR abnormalities in PSC as demonstrated by molecular and functional analyses and that these abnormalities may contribute to the development of PSC in a subset of patients with inflammatory bowel disease. PMID- 12783302 TI - Structural diversity and organization of three gene families for Kunitz-type enzyme inhibitors from potato tubers (Solanum tuberosum L.). AB - In the potato, Kunitz-type enzyme inhibitors are abundant and highly polymorphic small proteins found in tubers. DNA sequence analysis of 1596 unselected ESTs (expressed sequence tags) from mature tubers of the cultivars Provita and Saturna resulted in the identification of 55 different DNA sequences with high sequence similarity to Kunitz-type enzyme inhibitors. The frequency of Kunitz-type inhibitor ESTs in Provita was four times higher than in Saturna tubers, and none of the Provita ESTs was identical to any of the Saturna ESTs. A phenogram constructed from the deduced amino acid sequences of the inhibitors revealed three major homology groups-A, B and C. Group A inhibitors were all derived from Provita ESTs. Inhibitor groups A and B were more similar to each other than to group C inhibitors, and for most members within-group similarity was at least 90%. Non-conservative amino acid substitutions and insertion/deletion polymorphisms suggest functional differentiation between members of the gene family. A minimum of 21 genes for Kunitz-type enzyme inhibitors (six for group A, nine for group B and six for group C) was estimated to exist in the potato genome. Genetic mapping and the identification of BAC (bacterial artificial chromosome) clones containing more than one member of the gene family indicated that most inhibitor genes of groups A, B and C are organized in a cluster that maps to a single region on potato chromosome III. PMID- 12783303 TI - Functional comparison of homologous members of three groups of Kunitz-type enzyme inhibitors from potato tubers (Solanum tuberosum L.). AB - For functional studies, nine cDNAs encoding Kunitz-type enzyme inhibitors from potato tubers were expressed as GST (glutathione S transferase)-tagged fusion proteins in the fission yeast Schizosaccharomyces pombe. The inhibitors represented the three major homology groups A, B and C found in tubers. Members of the same homology group were at least 90% identical in sequence. The purified GST fusion proteins were tested for their ability to inhibit the proteases trypsin, alpha-chymotrypsin, subtilisin, papain and aspergillopepsin I, and for inhibition of the growth of fungi. Fusion proteins belonging to the same and different homology groups were found to exhibit distinct protease inhibition profiles. Removal of the GST tag by cleavage with enterokinase did not change the inhibition profile but increased the inhibitory activity. Group A and B inhibitors affected the proteases to different extents, whereas group C inhibitors showed only weak or no protease inhibition. One fusion protein completely inhibited aspergillopepsin I. One fusion protein each of groups A and B strongly inhibited mycelial growth of the fungus Fusarium moniliforme. The results suggest functional polymorphism among closely related members of the Kunitz-type inhibitor family. PMID- 12783304 TI - Genetic diversity of Cryptosporidium spp. in cattle in Michigan: implications for understanding the transmission dynamics. AB - Epidemiological and molecular data on 248 bovine, 17 human, and 16 water samples of Cryptosporidium spp. collected from the lower peninsula of Michigan between 1997 and 2000 were analysed. Cryptosporidium parvum bovine genotype and Cryptosporidium andersoni were found in 56 and four cattle samples, respectively. A total of six C. parvum subgenotypes were found in 34 bovine samples, and five of the eight farms had two or three subgenotypes in cattle. Six water samples from these farms had C. andersoni, five had the C. parvum bovine genotype, and one had Cryptosporidium muris. In contrast, four PCR-positive human samples produced the C. parvum bovine genotype and two had the C. parvum human genotype. Among the C. parvum bovine genotype samples, two human samples and one water sample had subgenotypes identical to those found on cattle farms. The results of this study demonstrate the potential use of molecular methods in tracking the transmission of Cryptosporidium. PMID- 12783305 TI - The role of Cryptosporidium parvum-derived phospholipase in intestinal epithelial cell invasion. AB - In the Cryptosporidium parvum-infected intestinal epithelial cell, the parasite occupies an unusual extracytoplasmic location at the luminal surface, but how the invading zoites interact with the host cell to achieve this niche is poorly understood. This study examined the role of secretory phospholipase A(2) (sPLA(2)), a known virulence factor for several pathogenic microorganisms, in establishing C. parvum intracellularly. Initially, it was established that there was sPLA(2) activity in homogenates of C. parvum oocysts. C. parvum reproduction in two human enterocyte cell lines was significantly reduced by a specific PLA inhibitor, p-bromophenacylbromide, and by sheep anti-sPLA(2) antibodies developed against PLA(2) of bee ( Apis mellifera) venom. Treatment of either C. parvum sporozoites or enterocytes with sPLA(2) derived from cobra ( Naja naja) venom before initiation of infection increased the numbers of intracellular parasites. Thus, C. parvum PLA(2 )may play an important part in establishing the parasite within the enterocyte. PMID- 12783306 TI - Canine echinococcosis in northern Jordan: increased prevalence and dominance of sheep/dog strain. AB - A total of 112 stray and semi-stray dogs (Canis familiaris) from four different geographical areas in northern and middle Jordan were necropsied to evaluate the prevalence and intensity of intestinal helminthiasis. Of these, 33 dogs (29.5%) were infected with Echinococcus granulosus and 61 (54.5%) with other Taenia species. Other cestodes found included Dipylidium caninum in 36 dogs (32.1%), Diplopylidium in 6 dogs (5.4%), Mesocestoides sp. in 3 dogs (2.7%) and Joyuexiella in 1 dog (0.9%). Toxocara nematodes were found in 10 dogs (9.2%) and only 1 dog was positive for acanthocephalans. Among the dogs infected with E. granulosus, 8 dogs (24.2%) had a worm load higher than 1,000 worms. The ratio of infected male to female dogs was 1.9:1.0. Strain analysis of E. granulosus using random primers revealed the dominance of the G1 strain (sheep/dog strain) in the region. Only one dog harbored another E. granulosus strain, which resembled the G4 strain pattern. PMID- 12783307 TI - Ultrastructural studies on the development of Enteromyxum scophthalmi (Myxozoa), an enteric parasite of turbot (Scophthalmus maximus L.). AB - The ultrastructure of the developmental stages of Enteromyxum scophthalmi is described. Scarce intracellular, early uninucleated stages appeared within intestinal epithelial cells whereas proliferative stages were abundant both intraepithelially and in the intestinal lumen. In the proliferative stages, food reserves were abundant in the cytoplasm of P cells and consisted mostly of carbohydrates in the intraepithelial stages and lipid inclusions in the luminal stages. Sporogenesis could occur in enveloped cells or by direct division or clustering of generative cells. The abundance, shape and size of mitochondria as well as the number and shape of their cristae were very variable in the different developmental stages. The cristae were usually tubular and sometimes plate-like, discoidal or lamellar. True flat cristae were not observed. We found elements of closed (cryptomitosis) and open mitosis as well as structures reminiscent of microtubule organising centres, hitherto not described in myxosporeans. The significance of these findings is discussed in relation to the taxonomic and phylogenetic position of the Myxozoa. PMID- 12783308 TI - Apoptosis induced by gamma irradiation of Taenia solium metacestodes. AB - Gamma irradiation of food is considered a possible approach to control food-borne diseases. In cysticercosis, previous studies have shown that irradiating (with 0.3 kGy) pork infected with Taenia solium larvae completely inhibits growth of the parasite. This study was conducted to evaluate the mechanisms that induce the effect of gamma irradiation on metacestodes of T. solium. Metacestodes were obtained from several infected pigs and irradiated with a dose of 0.3 kGy. The viability of the metacestodes was evaluated by their capacity to evaginate in vitro and in vivo development to tapeworms after they were orally infected into prednisolone-treated golden hamsters. Using the typical ladder pattern of fragmented DNA and the TdT-mediated DUTP-nick-end labeling assay, apoptosis was evaluated in metacestodes after irradiation and in the scolices and tapeworms recovered from infected hamsters at 21 days post-infection. Apoptosis was observed in the structure of scolices obtained from hamsters at 21 days post infection with irradiated metacestodes, This study provides evidence of the existence of apoptosis in the irradiated metacestodes of T. solium and helps elucidate the possible mechanisms that are involved when gamma irradiation inhibits the normal development of the T. solium metacestode into the adult worm. PMID- 12783309 TI - Potential role of head lice, Pediculus humanus capitis, as vectors of Rickettsia prowazekii. AB - Since the pioneering work of Charles Nicolle in 1909 [see Gross (1996) Proc Natl Acad Sci USA 93:10539-10540] most medical officers and scientists have assumed that body lice are the sole vectors of Rickettsia prowazekii, the aetiological agent of louse-borne epidemic typhus (LBET). Here we review the evidence for the axiom that head lice are not involved in epidemics of LBET. Laboratory experiments demonstrate the ability of head lice to transmit R. prowazekii, but evidence for this in the field has not been reported. However, the assumption that head lice do not transmit R. prowazekii has meant that head lice have not been examined for R. prowazekii during epidemics of LBET. The strong association between obvious (high) infestations of body lice and LBET has contributed to this perception, but this association does not preclude head lice as vectors of R. prowazekii. Indeed, where the prevalence and intensity of body louse infections may be high (e.g. during epidemics of LBET), the prevalence and intensity of head louse infestations is generally high as well. This review of the epidemiology of head louse and body louse infestations, and of LBET, indicates that head lice are potential vectors of R. prowazekii in the field. Simple observations in the field would reveal whether or not head lice are natural vectors of this major human pathogen. PMID- 12783310 TI - Amebic cysteine proteinase 2 (EhCP2) plays either a minor or no role in tissue damage in acute experimental amebic liver abscess in hamsters. AB - Amebic cysteine protease 2 (EhCP2) was purified from ethyl ether extracts of axenically grown trophozoites of Entamoeba histolytica strain HM1-IMSS. The purification procedure involved molecular filtration and electroelution. Sequence analysis of the purified product revealed EhCP2 and ubiquitin(s). Electrophoretic migration patterns, isoelectric point determination and Western blot studies failed to reveal other EhCP molecules. Polyclonal antibodies against the purified EhCP2 prepared in rabbits either stabilized or enhanced the enzyme activity in a dose-response manner. Purified EhCP2 was enclosed within inert resin microspheres (22-44 microm in diameter) and injected into the portal vein of normal hamsters. In the liver, the microspheres caused mild acute inflammation and occasional minimal necrosis of short duration. Sections of the liver were immunohistochemically stained with the anti-EhCP2 antibody and the microspheres were positive for only a very short period (1 h) after injection. Sections of experimental acute (1 day, 5 days) amebic liver abscess produced in hamsters were also stained with the anti-EhCP2 antibody; and amebas were intensely positive but no staining was observed at any time in the surrounding necrotic structures. It is suggested that EhCP2 plays either a minor or no role in the causation of tissue damage in experimental acute liver amebiasis. PMID- 12783311 TI - In vitro antimalarial activity of metalloporphyrins against Plasmodium falciparum. AB - The in vitro antimalarial activity against Plasmodium falciparum and heme polymerization were evaluated for ten metalloporphyrins: gallium protoporphyrin IX (GaPPIX), sodium salt of gallium protoporphyrin IX, silver protoporphyrin IX, palladium protoporphyrin IX, cobalt protoporphyrin IX, manganese protoporphyrin IX, tin protoporphyrin IX (SnPPIX), chromium protoporphyrin IX, gallium deuteroporphyrin IX (GaDPIX) and gallium hematoporphyrin IX. Metalloporphyrins inhibited parasite growth with 50% inhibitory concentrations (IC(50)) ranging from 15.5 microM to 190 microM. In trophozoite lysate-mediated heme polymerization assays, SnPPIX, GaPPIX and GaDPIX exerted potent inhibitory activity similar to that of artemisinin and chloroquine. PMID- 12783312 TI - Ultrastructure of Hepatozoon boigae (Mackerras, 1961) nov. comb. from brown tree snakes, Boiga irregularis, from northern Australia. AB - Intraerythrocytic bodies identified as haemogregarine gamonts were found in 29% of 97 brown tree snakes (Boiga irregularis) examined during a haematological survey of reptiles in Australasia during 1994-1998. The morphological characteristics of the parasites were consistent with those of Haemogregarina boigae Mackerras, 1961, although the gamonts were slightly larger and lacked red caps but contained distinctive polar grey capsules. Gamonts did not distend host cells but laterally displaced their nuclei. They were contained within parasitophorous vacuoles and possessed typical apicomplexan organelles, including a conoid, polar rings, rhoptries and micronemes. Schizonts producing up to 30 merozoites were detected in endothelial cells of the lungs of 11 snakes. The absence of erythrocytic schizogony suggests the parasites belong to the genus Hepatozoon. Electron microscopy also revealed the presence of curious encapsulated organisms in degenerating erythrocytes. These stages did not possess apical complex organelles and were surrounded by thick walls containing circumferential junctions and interposed strips reminiscent of oocyst sutures. PMID- 12783313 TI - Infectivity of Cryptosporidium parvum genotype I in conventionally reared piglets and lambs. AB - Parasites of the genus Cryptosporidium are intracellular parasites that occur throughout the animal kingdom and have been reported in many species of mammals, including human. Most infections in humans are caused by two C. parvum genotypes, genotype I and genotype II; these are the human and the bovine (zoonotic) genotypes, respectively. Successful experimental infection of Cryptosporidium parvum genotype I "human genotype" is described in four conventionally reared piglets and in a lamb. The inoculum was originally obtained from two diarrheic children, and the Cryptosporidium genotypes were determined by PCR and rDNA sequencing. The infective dose was between 10(6) and 2 x 10(6) oocysts. No clinical signs were observed in the infected animals, except in a piglet that showed watery diarrhea. The oocyst shedding period in positive animals ranged between 4 and 10 days. Histopathologic examination of the gastrointestinal tract of two positive piglets revealed shortening of the villi and denudation of the villous tips of the jejunum. In one piglet, the colon mucosa revealed numerous Cryptosporidium oocysts. The storage time of the inocula (< or =3 weeks in PBS at 4 degrees C) and the age of the animal (newborn) were important for the successful induction of infection. PMID- 12783314 TI - Taenia crassiceps cysticercosis: immune response in susceptible and resistant BALB/c mouse substrains. AB - Taenia crassiceps can naturally and experimentally infect rodents in which they reproduce by budding. Differences in the susceptibility to T. crassiceps cysticercosis were found between two BALB/c substrains: BALB/cAnN (susceptible) and BALB/cJ (resistant). In chimeric mice, resistance was transferred to susceptible mice with bone marrow cells from the resistant mice, which argues in favor of an immune mediation of the resistant phenotype. To further explore the immune response that could underlie these differences in susceptibility, the specific cellular immune response elicited by the parasite was explored in both substrains. An increased proliferative response and IL-2 levels were induced by cysticercal antigens only in splenocytes from resistant mice. A decrease in the percentage of CD4(+) (11.1%), CD8(+) (17.5%) was found in splenocytes from susceptible BALB/cAnN mice. A study of the TCRV beta repertoire revealed a significant decrease in V beta 2 in both CD4(+) and CD8(+) splenocytes only in the susceptible BALB/cAnN strain. PMID- 12783315 TI - Morphological studies on the extracellular structure of the midgut of a tick, Haemaphysalis longicornis (Acari: Ixodidae). AB - In the present study, morphological observations on the extracellular structures found on the apical surface of the midgut epithelium, known as the peritrophic membrane (PM) or glycocalyx, are described in Haemaphysalis longicornis females and larvae. These structures have been hypothesized to provide protection to the microvilli of epithelial cells of the digestive tract. Our aim was to determine whether the extracellular structures are important in the digestion of the blood meal and/or as a protection against infection or injury. The PM was detectable in the midgut of engorged larvae by electron microscopy, but not in engorged females. However, a PM-like structure, stainable with toluidine blue, was observed in females by light microscopy. From the results of confocal laser scanning and electron microscopic observations with wheat germ agglutinin (WGA lectin) staining for chitin of the PM, however, the structure was clearly recognized. The structure in the female is likely to be PM because staining with WGA lectin in the presence of GlcNAc indicates the presence of chitin and various morphologies of PM have been reported in insects and ticks. These results show morphologically that different types of PM-like structure are formed in larvae and females of H. longicornis. PMID- 12783316 TI - Electron microscopic identification of the intestinal protozoan flagellates of the xylophagous cockroach Parasphaeria boleiriana from Brazil. AB - Flagellate protozoa of the hindgut of the xylophagous blattid Parasphaeria boleiriana were examined by light and electron microscopy. This species harbours two oxymonad species of the genera Monocercomonoides and Polymastix, the latter bearing Fusiformis bacteria on its surface. A diplomonad was present and has features of the genus Hexamita rather than Spironucleus. In addition, two trichomonads of the genera Monocercomonas and Tetratrichomastix were identified. A precise comparison with species of blattids and other insects was difficult because most of these flagellates have been described only by light microscopy after cell staining and there are few electron microscope studies and no molecular studies. None of the flagellates contained wood fragments in their food vacuoles and so evidently do not participate in the digestion of wood or cellulose. PMID- 12783317 TI - Adenovirus-mediated prodrug-enzyme therapy for CEA-producing colorectal cancer cells. AB - PURPOSE: Carcinoembryonic antigen expression is increased in more than 80% of patients with colorectal cancer. Values are especially higher in patients with advanced stage disease. Virus directed prodrug/enzyme therapy (VDEPT) using genetically engineered viral vectors has been considered as one of the more notable cancer gene therapies for the transduction of various enzymes into cancer cells. We made adenovirus vectors under the control of a CEA promoter that included the HSV-tk gene and investigated its usefulness to specifically target human CEA-producing colorectal cancer cells. METHODS: An adenovirus vector with the lacZ or HSV-tk gene under the control of a CAG or CEA promoter was designed for the VDEPT experiment. Human colorectal cancer cell lines were used for in vitro experiments to assure the transduction efficacy of the inserted genes by these vectors. To conduct the in vivo experiment, liver metastases of the cell line were created in CB17 SCID mouse. We then performed intrasplenic injections of adenovirus vectors and intraperitoneal injections of the prodrug, ganciclovir. RESULTS: RCM-1, the CEA-producing human rectal cancer cell line, was more strongly stained by X-gal staining. Furthermore, COLO320 was faintly stained secondary to a shortage of CEA production. The in vivo VDEPT experiment with RCM 1 and the adenovirus vector driven by the CEA promoter revealed attenuation of liver metastases in the treatment group. CONCLUSIONS: Adenovirus vectors under the control of the CEA promoter can transduce inserted genes effectively into targeted human colorectal cancer cells according to the amount of expressed CEA protein. We anticipate the future use of VDEPT of the HSV-tk/GCV system using this vector in the treatment of advanced colorectal cancers. PMID- 12783319 TI - Epithelial cells in the oviduct and vagina and steroid-synthesizing cells in the rabbit ovary express AhR and ARNT. AB - The aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor. Beside exogenous ligands, an increasing list of endogenous ligands has recently been described. The AhR and its dimerization partner AhR nuclear translocator (ARNT) were found in embryos, fetuses and in genital tract tissue. Its role in reproduction and fertility is not known. In the current study, AhR and ARNT expression and co-localization were studied immunohistochemicaly during the pre implantation period in various genital tract organs (ovary, oviduct, vagina) of the rabbit. In the ovary, the AhR was expressed in the steroid-secreting interstitial cells, in follicular and granulosa cells, and in lutein cells. The receptor was localized in the cytoplasm. A cytoplasmic localization was also found in the oviduct epithelium with a diffuse cytoplasmic staining in the ampulla and a localized cytoplasmic localization in the isthmus. In the vagina, AhR localization changed from cytoplasmic in the non-pregnant animal to nuclear staining in the basal layer of the vaginal epithelium on day 6 of pregnancy. The ARNT protein was found in all AhR expressing cells except for oocytes within primordial follicles. Its localization was in the cytoplasm and/or nuclei. Therefore, the full complement for AhR/ARNT transcriptional activity was found in the studied organs. AhR expression showed stage-specific changes in both the uterus (Hasan and Fischer 2001) and the vagina during the pre-implantation period, while the ARNT protein exhibited no change in expression during this period. In summary, these findings indicate a functional AhR/ARNT complex in the rabbit genital tract epithelia. Its precise role and activation mechanism in reproductive tissue is not clear. It is supposed, however, that this complex may be involved in both hormone production and hormone-induced cellular changes during early pregnancy of the rabbit. PMID- 12783318 TI - Melatonin for treatment of sleeping disorders in children with attention deficit/hyperactivity disorder: a preliminary open label study. PMID- 12783320 TI - Developmental expression of glycocomponents in the chick chorioallantoic membrane. AB - Widespread interest has focused on the research of the chorioallantoic membrane (CAM) and its functional contribution to gaseous exchange, calcium reabsorption, water and electrolyte transport during chick embryogenesis. Nevertheless, very little information is available on the glycoconjugate components of this extra embryonic structure. In the present study, we investigated by lectin histochemistry, the glycosylation pattern expressed in the CAM epithelia during embryonic development. Occurrence of sialic acid-associated glycoproteins was detailed by either specific lectins, which discriminate alpha2,3 and alpha2,6 sialoderivatives, or sialidase digestion combined with appropriate lectins to identify the sialic acid acceptor sugars. Lectin affinities proved to depend greatly on differentiation of the CAM epithelia which showed highest expression of binding sites during the second half of incubation up to hatching. Differences emerged between the chorionic and the allantoic epithelium, regarding qualitative, quantitative and temporal expression of sugar moieties. A cell type specific distribution of glycocomponents was found in the chorionic epithelium where lectin binding sites were specifically located in the villus cavity cells. In the allantoic epithelium, high and heterogeneous occurrence of sialoglycoconjugates as well as specific presence of fucose residues were evidenced mostly in the granule cells. We conclude from these findings that various glycoconjugates in the CAM could participate in different physiological functions characteristic of the chorionic and the allantoic epithelium. PMID- 12783321 TI - Histological evaluation of age-related changes in human respiratory mucosa of the middle turbinate. AB - Though the lining of the healthy human nose is subject of numerous investigations, age-related changes have attracted little attention. Furthermore, varying sampling sites complicate the evaluation of the available information. Reference is needed for evaluation of abnormal conditions, e.g. in persons with sleep apnea. Biopsy samples from the middle turbinate of 40 healthy humans aged 5 to 75 years were examined. Tissue samples were fixed immediately in glutaraldehyde and embedded in Epon for semi-thin sectioning and blind histological evaluation. In all age groups, patchy arrangement of different epithelial types were found. Ciliated epithelium decrease with age. Stratified and atrophic epithelial types show striking changes. Thin atrophic epithelium with conspicuously thickened basement membranes is only seen above the age of 40 years and is frequently found in older subjects. Stratified epithelial types initially increase with age, but decrease above 50 years. In the middle aged group (26 to 50 years) we found a remarkably high amount of squamous metaplasia, resulting in overall thicker epithelium becoming thinner in older subjects. In contrast, basement membranes are thinnest in the middle aged group. In conclusion, evaluating the histology of nasal respiratory epithelium must take pronounced age-related physiological changes into account. PMID- 12783322 TI - Differential gene expression in the periprosthetic membrane: lubricin as a new possible pathogenetic factor in prosthesis loosening. AB - About 10% of hip endoprostheses will loosen after 10 years. Prosthesis loosening is caused by two different pathomechanisms: aseptic loosening (AL) and septic loosening (SL). This study evaluated differences in gene expression in AL and SL. Eight hybridizations were performed on PIQOR cDNA arrays. Objects of the study were periprosthetic interface tissue samples from two patients with SL and three patients with AL. Tissue parts directly adjacent to the site of RNA isolation were analyzed immuno/histopathologically in order to overcome the problem of tissue heterogeneity. Thirty-three genes were found constantly differentially expressed, among which were cd11b, cd18, cd68, osteopontin and ferritin heavy chain upregulated in AL and collagen types 1alpha-1, 3alpha-1, integrin alpha-1, thrombospondin2 and nidogen upregulated in SL. The most striking finding was the strong upregulation (from 20-fold to 323-fold) of megakaryocyte stimulating factor (msf) in all aseptic cases and one of the two septic cases, which was confirmed by real-time reverse transcription-polymerase chain reaction. In this study, msf is linked to prosthesis loosening for the first time. The upregulation in AL suggests an important pathogenetic role: the msf splice product lubricin is responsible for the lubrication of healthy joints, but its excellent lubrication ability may disturb the tight interaction between bone and prosthesis and thereby contribute to prosthesis loosening. PMID- 12783323 TI - Metastatic malignant melanoma with Homer-Wright rosettes mimicking a neuroblastic tumor. An unusual morphological manifestation. PMID- 12783324 TI - Epidermal growth factor receptor is a marker for syncytiotrophoblastic cells in testicular germ cell tumors. AB - The epidermal growth factor receptor (EGFR) has been implicated in the pathogenesis, therapy and prognosis of various tumor types. The aim of this study was to investigate EGFR expression in a large series of testicular germ cell tumors (TGCTs). A total of 88 TGCTs (37 of pure type and 51 of mixed type) comprising a total of 44 seminoma, 49 embryonal carcinoma, 32 yolk sac tumor, 28 teratoma and 7 choriocarcinoma components were immunostained for EGFR. EGFR reactivity was observed in the stromal cells of embryonal carcinoma (29%) and in epithelial compartments of teratoma (71%). In addition, EGFR staining was consistently detected in syncytiotrophoblastic cells of choriocarcinoma, seminoma, embryonal carcinoma and yolk sac tumor components. EGFR staining, similar to beta-human chorionic gonadotropin (HCG) immunohistochemistry, was efficiently able to identify syncytiotrophoblastic cells in TGCTs. This study shows that EGFR is expressed in a subset of testicular germ cell tumors and suggests that EGFR may be a useful marker for syncytiotrophoblastic cells. PMID- 12783325 TI - Interaction between proliferating cell nuclear antigen and JUN-activation-domain binding protein 1 in the meristem of rice, Oryza sativa L. AB - The eukaryotic polymerase processivity factor, proliferating cell nuclear antigen (PCNA), interacts with many cell cycle-regulator proteins and with proteins involved in the mechanisms of DNA replication and repair. In the present study using two-hybrid analysis with PCNA from rice, Oryza sativa L. cv. Nipponbare (OsPCNA), we found that OsPCNA interacted in vitro and in vivo with rice JUN activation-domain-binding protein 1 (OsJab1), which is known as COP9/signalsome subunit 5. Both OsPCNA and OsJab1 transcripts were expressed strongly in the shoot apical meristem and weakly in young leaves, flag leaves, ears, roots and root tips. No expression was detected in the mature leaves. The OsPCNA and OsJab1 proteins were expressed and accumulated mostly in the shoot apical meristem and ears, suggesting that OsJab1 is involved in cell proliferation in cooperation with OsPCNA. The role of OsPCNA with OsJab1 in plant DNA proliferation is discussed. PMID- 12783327 TI - Control of water uptake by rice ( Oryza sativa L.): role of the outer part of the root. AB - A new pressure-perfusion technique was used to measure hydraulic and osmotic properties of the outer part of roots (OPR) of 30-day-old rice plants (lowland cultivar: IR64, and upland cultivar: Azucena). The OPR comprised rhizodermis, exodermis, sclerenchyma and one cortical cell layer. The technique involved perfusion of aerenchyma of segments from two different root zones (20-50 mm and 50-100 mm from the tip) at precise rates using aerated nutrient solution. The hydraulic conductivity of the OPR (Lp(OPR)=1.2x10(-6) m s(-1) MPa(-1)) was larger by a factor of 30 than the overall hydraulic conductivity (Lp(r)=4x10(-8) m s(-1) MPa(-1)) as measured by pressure chamber and root pressure probe. Low reflection coefficients were obtained for mannitol and NaCl for the OPR (sigma(sOPR)=0.14 and 0.09, respectively). The diffusional water permeability ( P(dOPR)) estimated from isobaric flow of heavy water was smaller by three orders of magnitude than the hydraulic conductivity (Lp(OPR)/ P(fOPR)). Although detailed root anatomy showed well-defined Casparian bands and suberin lamellae in the exodermis, the findings strongly indicate a predominantly apoplastic water flow in the OPR. The Lp(OPR) of heat-killed root segments increased by a factor of only 2, which is in line with the conclusion of a dominating apoplastic water flow. The hydraulic resistance of the OPR was not limiting the passage of water across the root cylinder. Estimations of the hydraulic properties of aerenchyma suggested that the endodermis was rate-limiting the water flow, although the aerenchyma may contribute to the overall resistance. The resistance of the aerenchyma was relatively low, because mono-layered cortical septa crossing the aerenchyma ('spokes') short-circuited the air space between the stele and the OPR. Spokes form hydraulic bridges that act like wicks. Low diffusional water permeabilities of the OPR suggest that radial oxygen losses from aerenchyma to medium are also low. It is concluded that in rice roots, water uptake and oxygen retention are optimized in such a way that hydraulic water flow can be kept high in the presence of a low efflux of oxygen which is diffusional in nature. PMID- 12783326 TI - The small subunit ADP-glucose pyrophosphorylase ( ApS) promoter mediates okadaic acid-sensitive uidA expression in starch-synthesizing tissues and cells in Arabidopsis. AB - Transgenic plants of Arabidopsis thaliana Heynh., transformed with a bacterial beta-glucuronidase (GUS) gene under the control of the promoter of the small subunit (ApS) of ADP-glucose pyrophosphorylase (AGPase), exhibited GUS staining in leaves (including stomata), stems, roots and flowers. Cross-sections of stems revealed GUS staining in protoxylem parenchyma, primary phloem and cortex. In young roots, the staining was found in the root tips, including the root cap, and in vascular tissue, while the older root-hypocotyl axis showed prominent staining in the secondary phloem and paratracheary parenchyma of secondary xylem. The GUS staining co-localized with ApS protein, as found by tissue printing using antibodies against ApS. Starch was found only in cell and tissue types exhibiting GUS staining and ApS labelling, but not in all of them. For example, starch was lacking in the xylem parenchyma and secondary phloem of the root-hypocotyl axis. Sucrose potently activated ApS gene expression in leaves of wild-type (wt) plants, and in transgenic seedlings grown on sucrose medium where GUS activity was quantified with 4-methylumbelliferyl-beta-glucuronide as substrate. Okadaic acid, an inhibitor of protein phosphatases 1 and 2A, completely blocked expression of ApS in mature leaves of wt plants and prevented GUS staining in root tips and flowers of the transgenic plants, suggesting a similar signal transduction mechanism for ApS expression in various tissues. The data support the key role of AGPase in starch synthesis, but they also underlie the ubiquitous importance of the ApS gene for AGPase function in all organs/tissues of Arabidopsis. PMID- 12783329 TI - Down-regulation of the AtCCR1 gene in Arabidopsis thaliana: effects on phenotype, lignins and cell wall degradability. AB - Cinnamoyl CoA reductase (CCR; EC 1.2.1.44) is the first enzyme specific to the biosynthetic pathway leading to monolignols. Arabidopsis thaliana (L.) Heynh. plants transformed with a vector containing a full-length AtCCR1 cDNA in an antisense orientation were obtained and characterized. The most severely down regulated homozygous plants showed drastic alterations to their phenotypical features. These plants had a 50% decrease in lignin content accompanied by changes in lignin composition and structure, with incorporation of ferulic acid into the cell wall. Microscopic analyses coupled with immunolabelling revealed a decrease in lignin deposition in normally lignified tissues and a dramatic loosening of the secondary cell wall of interfascicular fibers and vessels. Evaluation of in vitro digestibility demonstrated an increase in the enzymatic degradability of these transgenic lines. In addition, culture conditions were shown to play a substantial role in lignin level and structure in the wild type and in the effects of AtCCR1 repression efficiency. PMID- 12783328 TI - Elevated levels of N-lauroylethanolamine, an endogenous constituent of desiccated seeds, disrupt normal root development in Arabidopsis thaliana seedlings. AB - N-Acylethanolamines (NAEs) are prevalent in desiccated seeds of various plant species, and their levels decline substantially during seed imbibition and germination. Here, seeds of Arabidopsis thaliana (L.) Heynh. were germinated in, and seedlings maintained on, micromolar concentrations of N-lauroylethanolamine (NAE 12:0). NAE 12:0 inhibited root elongation, increased radial swelling of root tips, and reduced root hair numbers in a highly selective and concentration dependent manner. These effects were reversible when seedlings were transferred to NAE-free medium. Older seedlings (14 days old) acclimated to exogenous NAE by increased formation of lateral roots, and generally, these lateral roots did not exhibit the severe symptoms observed in primary roots. Cells of NAE-treated primary roots were swollen and irregular in shape, and in many cases showed evidence, at the light- and electron-microscope levels, of improper cell wall formation. Microtubule arrangement was disrupted in severely distorted cells close to the root tip, and endoplasmic reticulum (ER)-localized green fluorescent protein (mGFP5-ER) was more abundant, aggregated and distributed differently in NAE-treated root cells, suggesting disruption of proper cell division, endomembrane organization and vesicle trafficking. These results suggest that NAE 12:0 likely influences normal cell expansion in roots by interfering with intracellular membrane trafficking to and/or from the cell surface. The rapid metabolism of NAEs during seed imbibition/germination may be a mechanism to remove this endogenous class of lipid mediators to allow for synchronized membrane reorganization associated with cell expansion. PMID- 12783331 TI - The root epidermis of Echium plantagineum L.: a novel type of pattern based on the distribution of short and long root hairs. AB - The great majority of angiosperm species form a group in which either every cell in the root epidermis produces a root hair, or the cells that produce these hairs are randomly distributed. We describe, for the first time, pattern in the root epidermal cells of a species within this group. The seedling root of Echium plantagineum L. (Boraginaceae) has an epidermis in which almost every cell produces a root hair, but these are of two types, short hairs (up to 200 micro m) and long hairs (>200 micro m), which are in separate cell files, with the cells bearing long hairs usually separated by one or two files of cells bearing short hairs; the epidermal cells with the long root hairs are longer than the epidermal cells with the short root hairs. The long root hairs are initiated and develop earlier than the short root hairs. Transverse sections of the region of the root which contains only developing long root hairs show that the hair cells are located above anticlinal walls between underlying cortical cells. We regard the distribution of root epidermal cells in E. plantagineum as a sub-type of this group. We discuss the possible evolution, from this sub-type, of another group that is characterised by hair cells and non-hair cells occurring in separate files. PMID- 12783330 TI - Localization of dibenzodioxocin substructures in lignifying Norway spruce xylem by transmission electron microscopy-immunogold labeling. AB - The lignification process in mature Norway spruce [ Picea abies (L.) H. Karsten] xylem cell walls was studied using transmission electron microscopy (TEM) immunogold detection with a polyclonal antibody raised against a specific lignin substructure, dibenzodioxocin. The study reveals for the first time the exact location of this abundant eight-ring structure in the cell wall layers of wood. Spruce wood samples were collected in Southern Finland at the time of active growth and lignification of the xylem cell walls. In very young tracheids where secondary cell wall layers were not yet formed, the presence of the dibenzodioxocin structure could not be shown at all. During secondary cell wall thickening, the dibenzodioxocin structure was more abundant in the secondary cell wall layers than in the middle lamella. The highest number of gold particles revealing dibenzodioxocin was in the S2+S3 layer. Statistically significant differences were found in the frequency of gold particles present in various cell wall layers. For comparison, wood sections were also cut with a cryomicrotome for light and fluorescence microscopy. PMID- 12783332 TI - Involvement of sulfoquinovosyl diacylglycerol in the structural integrity and heat-tolerance of photosystem II. AB - To examine the role of sulfoquinovosyl diacylglycerol (SQDG) in thylakoid membranes, we compared the structural and functional properties of photosystem II (PSII) between a mutant of Chlamydomonas reinhardtii defective in SQDG ( hf-2) and the wild type. The PSII core complex of hf-2, as compared with that of the wild type, showed structural fragility when solubilized with a detergent, dodecyl beta- d-maltoside, suggesting that the physical properties of the PSII complex were altered by the loss of SQDG. On the other hand, exposure of the cells to 41 degrees C for 120 min in the dark decreased the PSII activity to 70% and 50% of the initial levels in the wild type and hf-2, respectively, which implies that the PSII activity, in the absence of SQDG, becomes less stable under heat-stress conditions. PSII inactivated to 60% of the initial level by dark incubation at 41 degrees C was reactivated by following illumination even at 41 degrees C to more than 90% in the wild type, but only to 70% in hf-2. These results suggest that PSII inactivated by heat recovers through some mechanism dependent on light, and that SQDG participates in functioning of the mechanism. The conformational disorder of PSII caused by the defect in SQDG might be correlated with the increased susceptibility of its activity to heat-stress. PMID- 12783333 TI - Localization of sucrose synthase in wheat roots: increased in situ activity of sucrose synthase correlates with cell wall thickening by cellulose deposition under hypoxia. AB - Sucrose synthase (SuSy; EC 2.4.1.13) plays a prominent role in O(2) deficiency and functions at a branch point, partitioning sucrose between cell wall biosynthesis and glycolysis. The cleavage of sucrose by SuSy was localized in wheat ( Triticum aestivum L. cv. Alcedo) roots subjected to 4 days of hypoxia. Increased SuSy activity was observed by in situ activity staining in the tip region and in the stele of root axes. The pattern of cellulose deposition correlated with regions of high SuSy activity. Cellulose accounted for more than 30% of root dry weight and the cellulose content increased substantially under hypoxia. The strongest accumulation of cellulose occurred in the base and mid regions of the roots where the content rose to 163% and 182% of controls, respectively. In the root axis, cellulose deposition occurred in the endodermis and walls of pith cells. In root tips, cellulose was primarily deposited in developing xylem and phloem. The marker enzyme for O(2) shortage, pyruvate decarboxylase (EC 4.1.1.17), exhibited a 14-fold increase in the root apex, whereas in basal root tissues, which contained more aerenchyma, pyruvate decarboxylase activity was only doubled. The root apex also contained the highest concentration of sucrose and hexoses. The elevated sugar content in all root zones was partially used to synthesize cellulose for secondary wall thickening. PMID- 12783334 TI - Vernalization-induced flowering in wheat is mediated by a lectin-like gene VER2. AB - A vernalization-related gene VER2 was isolated from winter wheat ( Triticum aestivum L.) using a differential screening approach. The deduced VER2 is a lectin-like protein of 300 amino acids, which contains the presence of a jacalin like GWG domain. RNA in situ hybridization results demonstrated that VER2 gene expression is restricted to the marginal meristems of immature leaves in vernalized wheat seedlings. No hybridization signal was detected in the epidermal tissue and vascular bundles. However, "devernalization" resulted in the silencing of VER2 gene activity. The gene expression pattern of VER2 induced by jasmonate was similar to that induced by vernalization. Antisense inhibition of VER2 in transgenic wheat showed that heading and maturation time were delayed up to 6 weeks compared with non-transformed wheat and the pBI121empty-vector-transformed wheat. Tissue degeneration at the top of the spike was also noticed in the antisense inhibited transgenic wheat. These results suggest that VER2 plays an important role in vernalization signaling and spike development in winter wheat. PMID- 12783335 TI - In vitro biosynthesis of galactans by membrane-bound galactosyltransferase from radish ( Raphanus sativus L.) seedlings. AB - We investigated a galactosyltransferase (GalT) involved in the synthesis of the carbohydrate portion of arabinogalactan-proteins (AGPs), which consist of a beta (1-->3)-galactan backbone from which consecutive (1-->6)-linked beta-Gal p residues branch off. A membrane preparation from 6-day-old primary roots of radish ( Raphanus sativus L.) transferred [(14)C]Gal from UDP-[(14)C]Gal onto a beta-(1-->3)-galactan exogenous acceptor. The reaction occurred maximally at pH 5.9-6.3 and 30 degrees C in the presence of 15 mM Mn(2+) and 0.75% Triton X-100. The apparent K(m) and V(max) values for UDP-Gal were 0.41 mM and 1,000 pmol min( 1) (mg protein)(-1), respectively. The reaction with beta-(1-->3)-galactan showed a bi-phasic kinetic character with K(m) values of 0.43 and 2.8 mg ml(-1). beta-(1 ->3)-Galactooligomers were good acceptors and enzyme activity increased with increasing polymerization of Gal residues. In contrast, the enzyme was less efficient on beta-(1-->6)-oligomers. The transfer reaction for an AGP from radish mature roots was negligible but could be increased by prior enzymatic or chemical removal of alpha- l-arabinofuranose (alpha- l-Ara f) residues or both alpha- l Ara f residues and (1-->6)-linked beta-Gal side chains. Digestion of radiolabeled products formed from beta-(1-->3)-galactan and the modified AGP with exo-beta-(1- >3)-galactanase released mainly radioactive beta-(1-->6)-galactobiose, indicating that the transfer of [(14)C]Gal occurred preferentially onto consecutive (1-->3) linked beta-Gal chains through beta-(1-->6)-linkages, resulting in the formation of single branching points. The enzyme produced mainly a branched tetrasaccharide, Galbeta(1-->3)[Galbeta(1-->6)] Galbeta(1-->3)Gal, from beta-(1- >3)-galactotriose by incubation with UDP-Gal, confirming the preferential formation of the branching linkage. Localization of the GalT in the Golgi apparatus was revealed on a sucrose density gradient. The membrane preparation also incorporated [(14)C]Gal into beta-(1-->4)-galactan, indicating that the membranes contained different types of GalT isoform catalyzing the synthesis of different types of galactosidic linkage. PMID- 12783336 TI - Atomic force microscopy of microfibrils in primary cell walls. AB - Examination of angiosperm primary cell walls by transmission electron microscopy shows that they contain microfibrils that probably consist of cellulose microfibrils surrounded by associated non-cellulosic polysaccharides. Previous studies using solid-state (13)C NMR spectroscopy have shown that the cellulose is all crystalline with crystallites of cross-sectional dimensions of 2-3 nm. However, it is not known if each microfibril contains only one, or more than one crystallite because there is no agreement about the dimensions of the microfibrils. Partially hydrated primary cell walls isolated from onion ( Allium cepa L.) and Arabidopsis thaliana (L.) Heynh. were examined by atomic force microscopy and the microfibril diameters determined. The cell walls of both species contained tightly interwoven microfibrils of uniform diameter: 4.4+/-0.13 nm in the onion and 5.8+/-0.17 nm in A. thaliana. The effect was also examined of extracting the A. thaliana cell walls to remove pectic polysaccharides. The microfibrils in the extracted cell walls of A. thaliana were significantly narrower (3.2+/-0.13 nm) than those in untreated walls. The results are consistent with the microfibrils containing only one cellulose crystallite. PMID- 12783337 TI - Enhancement of cold tolerance and inhibition of lipid peroxidation by citrus dehydrin in transgenic tobacco. AB - Citrus ( Citrus unshiu Marcov.) dehydrin in response to chilling stress was overexpressed in tobacco ( Nicotiana tabacum L.), and the cold stress tolerance of transgenics at low temperature was analyzed. The freezing at -4 degrees C for 3 h of 24 independent lines indicated that a phenotype expressing citrus dehydrin showed less electrolyte leakage than the control. Dehydrin protein content was correlated with freezing tolerance in transgenics. Dehydrin-expressing tobacco exhibited earlier germination and better seedling growth than the control at 15 degrees C. Cell fractionation experiments suggested that the protein was predominantly expressed in mitochondria and the soluble fraction. Malondialdehyde production enhanced by chilling stress was lower in tobacco plants expressing citrus dehydrin than in control phenotypes. Dehydrin protein, purified from Escherichia coli expressing citrus dehydrin cDNA, prevented peroxidation of soybean ( Glycine max L.) liposomes in vitro. The inhibitory activity of dehydrin against liposome oxidation was stronger than that of albumin, glutathione, proline, glycine betaine, and sucrose. These results suggest that dehydrin facilitates plant cold acclimation by acting as a radical-scavenging protein to protect membrane systems under cold stress. PMID- 12783338 TI - Two Brassica napus polygalacturonase inhibitory protein genes are expressed at different levels in response to biotic and abiotic stresses. AB - Plants encode a distinct set of polygalacturonase inhibitory proteins (PGIPs) that function to inhibit polygalacturonase enzymes produced by soft-rot fungal pathogens. We characterized two PGIP-encoding genes ( Bnpgip1 and Bnpgip2) from Brassica napus DH12075 (a double-haploid line derived from a cross between 'Cresor' and 'Westar'). The two proteins exhibit 67.4% identity at the amino acid level and contain 10 imperfect leucine-rich repeats. The pgip genes are present as a small multigene family in B. napus with at least four members. Bnpgip1 and Bnpgip2 are constitutively expressed in roots, stems, flower buds and open flowers. In mature leaf tissue, different levels of induction were observed in response to biotic and abiotic stresses. Bnpgip1 expression was highly responsive to flea beetle feeding and mechanical wounding, weakly responsive to Sclerotinia sclerotiorum infection and exposure to cold but not to dehydration. Conversely, Bnpgip2 expression was strongly induced by S. sclerotiorum infection and to a lesser degree by wounding but not by flea beetle feeding. Application of jasmonic acid to leaves induced both Bnpgip1 and Bnpgip2 gene expression; however, salicylic acid did not activate either gene. Taken together, these results suggest that separate pathways regulate Bnpgip1 and Bnpgip2, and that their roles in plant development or resistance to biotic and abiotic stress differ. PMID- 12783340 TI - Changes to the galactose/mannose ratio in galactomannans during coffee bean ( Coffea arabica L.) development: implications for in vivo modification of galactomannan synthesis. AB - Endosperm was isolated from Arabica Caturra coffee beans 11, 15, 21, 26, 31 and 37 weeks after flowering, and the chemical composition and relative solubility of its component polysaccharides determined at each growth stage. Chemical analysis of the total mannan content of the cell wall material was done after solubilisation of galactomannans by alkaline extraction of the cell wall material followed by enzymatic digestion of the alkali-insoluble residue with a mixture of endo-mannanse and endo-glucanase. Eleven weeks after flowering, galactomannans accounted for approximately 10% of the polysaccharides but were highly substituted, with galactose/mannose ratios between 1:2 and 1:7. As the bean matured, galactomannan became the predominant polysaccharide, until 31 weeks after flowering it accounted for approximately 50% of the polysaccharides. However, it was less substituted, possessing galactose/mannose ratios between 1:7 and 1:40. Early in bean growth, up to 50% of the cell wall polysaccharides were extractable but as the galactomannan content of the bean increased there was a reduction in the extractability of all polysaccharides. The decrease in the galactose/mannose ratio of the galactomannans commenced between 21 and 26 weeks after flowering and was in synchrony with a rise in the concentration of free galactose in the beans. The results indicated that the degree of substitution of the galactomannans in coffee beans is developmentally regulated and may result, in part, from the modification of a primary synthetic product by the action of an alpha-galactosidase. PMID- 12783342 TI - Improvement of the automatic endoscopic reprocessor: self-cleaning disinfecting connectors between endoscope and reprocessor. AB - BACKGROUND AND STUDY AIMS: We have previously pointed out a defect of automatic endoscopic reprocessors, i. e. the contamination of the connecting part between the endoscope and the reprocessor. We evaluated a newly designed connector (MH 861; Olympus, Tokyo, Japan) with a self-cleaning and disinfection mechanism, which enabled cleaning and disinfection of both the connector itself and its interface with the suction and air/water valves during a reprocessing cycle, which was not previously possible. METHODS: Ten upper gastrointestinal endscopes were examined in the study. Swabs were taken from the suction and air/water valves for microbiological culture before and after reprocessing by the washer disinfector. The numbers of contaminated endoscopes before and after reprocessing with the new connector were compared. RESULTS: Before the procedure there were five contaminated endoscopes and none after the procedure. When the new connector was used, the difference in cleaning and disinfection of the connecting parts was significant (P = 0.0325). CONCLUSIONS: We conclude that the newly developed connector permits effective cleaning and disinfection by automatic reprocessors. PMID- 12783339 TI - Regulation of amino acid uptake by carbon and nitrogen in Pinus sylvestris. AB - The simultaneous uptake of 13 different amino acids by Scots pine ( Pinus sylvestris L.) was characterized and its regulation investigated after pre treatments with a range of C and N metabolites. The uptake system exhibited a broad substrate specificity, acquiring all tested amino acids at similar uptake rates. Uptake of all tested amino acids by excised roots was linear over a time period of 150 min and exhibited pH-dependency, showing a peak at pH 4.0-5.0. Uptake was increased following pre-treatments with glucose and sucrose, while ammonium pre-treatments had a negative effect on amino acid acquisition. Pre treatments with the important Krebs cycle intermediates oxaloacetate or 2 oxoglutarate did not result in altered amino acid uptake. It is speculated that the up-regulation of uptake may be due to an increased flow of glucose through a sensor mechanism, such as hexokinase. The regulation of transport by N and C suggests a function for amino acid uptake in the N nutrition of the plant, thus further highlighting the importance of organic nitrogen for plant N nutrition suggested by an increasing amount of research. PMID- 12783343 TI - Reduced risk for pancreatitis after endoscopic microtransducer manometry of the sphincter of Oddi: a randomized comparison with the perfusion manometry technique. AB - BACKGROUND AND STUDY AIMS: Endoscopic microtransducer manometry of the sphincter of Oddi has been shown to be a reliable alternative to perfusion manometry for evaluating sphincter of Oddi motor function. It avoids volume loading of the biliopancreatic system, and may therefore be associated with a lower risk of inducing postmanometry pancreatitis.[nl] PATIENTS AND METHODS: During a 2-year period, microtransducer manometry of the sphincter of Oddi was carried out in 215 patients (median age 42 years; 155 women; biliary study in 152 cases, additional pancreatic evaluation in 63 cases). Sphincter of Oddi manometry was conducted as the sole procedure in all patients. The frequency of pancreatitis was assessed prospectively and graded according to established guidelines. A total of 130 consecutive patients (median age 37 years, 92 women; 30 with biliary type II, 58 with type III, 34 with pancreatic type I, and eight with type II) were then randomly assigned to undergo microtransducer or perfusion manometry of the sphincter of Oddi in a standardized fashion. RESULTS: During the initial 2-year period, nine cases of pancreatitis (a pancreatitis frequency of 4.2 %) were observed after microtransducer manometry, and most were of mild degree (six mild, two moderate, and one severe). No deaths occurred, and no surgical procedures were required. In the randomized part of the study, the demographic and clinical characteristics of the patients in both groups, as well as the technical aspects of the procedures performed, were well matched. The frequency of pancreatitis after microtransducer manometry was 3.1 %, compared with 13.8 % after perfusion manometry ( P < 0.05). Pancreatitis occurred in two patients after microtransducer manometry, and was mild in both cases. After perfusion manometry, mild pancreatitis occurred in six patients and moderate pancreatitis in three. CONCLUSION: Endoscopic microtransducer manometry of the sphincter of Oddi is associated with a lower risk for postmanometry pancreatitis than standard perfusion manometry. PMID- 12783344 TI - A prospective randomized study of hydrophilic polymer-coated polyurethane versus polyethylene stents in distal malignant biliary obstruction. AB - BACKGROUND AND STUDY AIMS: Hydrophilic polymer-coated polyurethane (HPCP) stents have a low friction coefficient and a hydrophilic layer, which may reduce biofilm formation and increase the period of stent patency. We compared the patency rates with this new stent with the standard Amsterdam-type polyethylene (PE) stent in a prospective randomized trial. PATIENTS AND METHODS: One hundred patients with an unresectable distal malignant bile duct stricture without a previous drainage procedure were randomly assigned to receive either a HPCP stent or a PE stent. The diameter (10 Fr), length (9 cm) and stent design (Amsterdam type) were similar in both stents. Nine patients were excluded. Forty-four patients received an HPCP stent and 47 patients a PE stent. The diagnoses included carcinoma of the pancreas (n = 78), papilla (n = 1), bile duct (n = 10), and metastases (n = 2). RESULTS: Stent insertion was successful in all patients. Stent dysfunction occurred in 27 of the HPCP stents and 20 of the PE stents, with median stent patency periods of 77 days (95 % CI, 53-101 days) for HPCP stents and 105 days (95 % CI, 42-168 days) for PE stents. The patency period was significantly longer for the PE stent (P = 0.04). Early complications occurred in four patients (4%), one in the HPCP group and three in the PE group. CONCLUSION: Hydrophilic polymer coated polyurethane stents do not prolong the patency period of biliary stents. In fact, the current standard treatment using polyethylene stents in patients with distal malignant biliary obstruction showed a significantly longer patency period. PMID- 12783345 TI - Endoscopic palliation of malignant gastric outlet obstruction using self expandable metallic stents: results of a multicenter study. AB - BACKGROUND AND STUDY AIMS: Gastric outlet obstruction is a late event in the natural history of biliopancreatic tumours. Metallic self-expanding stents inserted under endoscopic and fluoroscopic guidance can be used for palliation. The aim of this study was to evaluate the feasibility, efficacy, and complications of endoscopic duodenal stenting in patients with malignant gastric outlet obstruction. PATIENTS AND METHODS: Between August 1998 and November 2001, 63 patients (31 women, 32 men; mean age 73 +/- 12) presenting with clinical symptoms of duodenal obstruction underwent endoscopic stenting with large metallic prostheses. Complications and clinical outcome were assessed both retrospectively and prospectively. RESULTS: Of the patients, 58 needed one duodenal stent and two overlapping stents were required in five patients. Stenting was immediately successful in 60/63 patients (95%). At the time of the duodenal procedure, 25 previously inserted biliary stents were still patent; biliary stenting was attempted during the same procedure in 18 patients; and 20 patients had no biliary stricture. There was no procedure-related mortality. There were complications in 30 % of patients: 13 stent obstructions, 4 stent migrations and 2 duodenal perforations (treated surgically). For 44 patients (70%) there were no minor or major digestive problem during their remaining lifetime. An exclusively peroral diet was possible in 58 patients (92%), but was considered satisfactory (solid or soft) in 46/63 patients (73%). Of the patients, 53 (84 %) died between 1 and 64 weeks after the duodenal stenting (median survival 7 weeks). CONCLUSIONS: Endoscopic stenting for the palliation of malignant gastric outlet obstruction is feasible and well-tolerated in most patients. Most dysfunctions can be managed endoscopically. PMID- 12783346 TI - Long-term results of balloon catheter dilation for benign gastric outlet stenosis. AB - BACKGROUND AND STUDY AIMS: In 1981, the authors introduced balloon catheter dilation for postoperative gastric outlet stenosis and later for peptic, corrosive and postvagotomy gastric outlet stenosis. This retrospective study evaluates the effectiveness, safety and outcome of balloon catheter dilation in these various indications. PATIENTS AND METHODS: Between September 1981 and September 2001, 177 balloon catheter dilation procedures were carried out in 72 patients with benign stenoses. Double-lumen or single-lumen balloon catheters with a mean diameter of 18 mm (range 12-25 mm) were used. Endoscopic check-up examinations were carried out 1-3 weeks later, and then after three, six and 12 months, or if symptoms returned. The mean follow-up period for patients who did not undergo surgery was 98 months (range 12-240 months). RESULTS: Symptomatic relief was obtained immediately in 80 % and after 3 months or more in 70% of the patients. The mean diameter of the stenoses was 6 mm (2.0-9.5 mm) before dilation and 16 mm (10-20 mm) afterwards. Gastric retention was observed in 49 patients (68%) before dilation and in 19 patients (26.4%) afterward. Sixteen patients had recurrent stenosis 1-18 months after the first dilation. All of the 18 postoperative strictures, 21 (70%) of the 30 peptic stenoses, six (35%) of the 17 patients with corrosive strictures, and five of the six patients with postvagotomy functional stenosis were successfully treated with dilation. Pyloric perforation occurred in two cases, and arterial hemorrhage was observed in one case after dilation. CONCLUSIONS: Balloon catheter dilation is an important and effective diagnostic and therapeutic method; depending on the causative factor, it can make surgery unnecessary in nearly 70% of patients with benign gastric outlet stenosis. PMID- 12783347 TI - Eradication of dysplastic Barrett's oesophagus using photodynamic therapy: long term follow-up. AB - BACKGROUND AND STUDY AIMS: Barrett's oesophagus is a major risk factor for oesophageal adenocarcinoma, a condition which is rapidly increasing in incidence. Photodynamic therapy (PDT) is a developing treatment in which tissue damage is caused by the action of light on a previously administered photosensitizing agent. We present the results of long-term follow-up of its efficacy in patients with dysplastic Barrett's oesophagus. PATIENTS AND METHODS: A total of 40 patients with low-grade dysplasia in Barrett's oesophagus were treated with oral 5-aminolaevulinic acid (ALA) at a dose of 30 mg/kg, followed by laser endoscopy 4 hours later. Patients were treated between December 1995 and December 1998, and all were followed up regularly with endoscopy and biopsies in our surveillance programme. RESULTS: Among the patients, 35 (88%) showed a macroscopic reduction in the area of the columnar epithelium, and in all 40 patients dysplasia was found to be eradicated at 1 month. The effect has been maintained for a median follow-up of 53 months (range 18-68 months), although one patient developed a late carcinoma in an untreated area of Barrett's oesophagus 3 years after the intervention. CONCLUSIONS: Safe and effective ablation of low-grade dysplastic Barrett's oesophagus can be achieved with the use of ALA-induced PDT, and the effects are maintained in the long term. PMID- 12783348 TI - Endoscopic findings in end-stage renal disease. AB - BACKGROUND AND STUDY AIMS: Patients with end-stage renal disease (ESRD) may demonstrate a number of gastrointestinal lesions and suffer subsequent complications. Our aim was to investigate the endoscopic findings in these patients and identify the predisposing factors. PATIENTS AND METHODS: During a 1 year period (February 2000 to January 2001), we studied consecutive patients with end-stage renal disease who were undergoing routine endoscopy before renal transplantation. The rapid urease test was also performed to detect Helicobacter pylori infection. Demographic and clinical data were collected. Logistic regression analysis was used to determine the risk factors for important endoscopic lesions, including esophagitis, gastroduodenal erosions, and peptic ulcers. RESULTS: We studied 206 patients (124 male, 82 female, mean age 38.9). Of the patients 73.8% were asymptomatic but some patients experienced nausea (12.6%), heartburn (8.7%), and abdominal pain (7.3%). Endoscopy was normal in 74 patients (35.9%). Abnormal endoscopic findings were duodenal erosions (32.0%), antral erosions (22.8%), diffuse antral erythema (27.8%), duodenal ulcer (7.3%), esophagitis (5.8%), angiodysplasia (4.4%), nodular duodenum (2%), and inflammatory gastric polyps (1.5%). The rapid urease test was positive in 58.8% of patients. Important endoscopic lesions were more common in men and in H. pylori-infected patients. Age, duration of dialysis, cause of the ESRD, presence of any symptoms, and hemoglobin levels were not found to be related to these lesions. Most patients with peptic ulcers were asymptomatic. CONCLUSIONS: Duodenal erosions (32.0%), gastric erosions (22.8%), diffuse antral erythema (27.8%) and duodenal ulcer (7.3%) are common lesions in patients with end-stage renal disease. Male gender and H. pylori infection are associated with a higher risk of these lesions. As there is no association between patients' symptoms and gastroduodenal lesions, which may increase the risk of post-transplant complications, the development of diagnostic strategies for the detection of these lesions is recommended. PMID- 12783349 TI - Invasive colorectal cancer detected up to 3 years after a colonoscopy negative for cancer. AB - BACKGROUND AND STUDY AIMS: Colonoscopy has replaced barium enema as the primary method for direct diagnosis of colorectal cancer, but detection may fail, and the reasons for this are not completely understood. PATIENTS AND METHODS: In order to analyze the accuracy of colonoscopy for detecting invasive colorectal cancer, 7365 colonoscopic examinations were matched with the most accurate local government population-based cancer registry in Japan. RESULTS: In 15 colonoscopic examinations, patients were not diagnosed as having invasive colorectal cancer, but disease of this type was detected within 3 years of the examinations (false negative examinations). During the same period, 233 colonoscopies were identified as true-positive examinations. The false-negative rate for detecting invasive colorectal cancer with colonoscopy was 6% at 3 years. The false-negative rate was significantly higher in individuals between 60 and 69 years of age and in invasive cancers located to the right of the splenic flexure. CONCLUSIONS: Colonoscopists should receive adequate training in achieving easy cecal intubation, detecting small or flat lesions, and carrying out adequate biopsies. PMID- 12783350 TI - Drainage of deep pelvic abscesses using therapeutic echo endoscopy. AB - BACKGROUND AND STUDY AIMS: The purpose of this study was to evaluate the clinical efficacy of endosonographically guided transrectal aspiration and drainage by plastic stent of deep pelvic abscesses, using a therapeutic echo endoscope device. PATIENTS AND METHODS: Between September 2000 and June 2001, 12 patients (nine men, three women, mean age 67 years) were treated for a perirectal or a pelvic abscess using an endoscopic ultrasound (EUS) technique. The drainage of these fluid collections was performed under EUS guidance, using therapeutic EUS scopes with a large working channel. RESULTS: No major complication occurred during this study. Transrectal stent insertion succeeded in nine patients. In three patients, only aspiration was possible. Among the nine patients in whom a stent was successfully introduced into the fluid collection, complete drainage without relapse was achieved in eight patients (mean follow-up 10.6 months, range 6-14 months). The stent was removed endoscopically after 3 to 6 months. Drainage was incomplete in one patient (with a large abscess, diameter > 8 cm), who subsequently underwent surgical drainage. However, two out of the three patients in whom aspiration alone was performed developed a recurrence of the abscess and required surgical treatment. CONCLUSION: EUS-guided drainage of deep pelvic abscesses could offer an alternative treatment to surgery in the management of these postoperative complications. PMID- 12783351 TI - Score card endoscopy: a multicenter study to evaluate learning curves in 1-week courses using the Erlangen Endo-Trainer. AB - BACKGROUND AND STUDY AIMS: The present study was carried out in the context of current discussions concerning ways in which simulation systems can be integrated sensibly and effectively into clinical educational structures, in order to shorten the training period for assistants and reduce potential risks for the patient. In our study, a number of centers used a standardized training approach, in 1 week courses, to investigate the learning curve improvement that can be achieved with a group of beginners in upper gastroduodenal endoscopy. MATERIALS AND METHODS: The multicenter study used the Erlangen Endo-Trainer, with specially prepared biological specimens from pigs. Using this, the individual steps of diagnostic upper gastrointestinal endoscopy with biopsy can be carried out following a score-card system. After a theoretical introduction and a demonstration of the examination by an experienced endoscopist, an initial evaluation score for each participant was obtained on day 1. On the following days, the program consisted of 2 hours' training by a tutor, followed by a test run for each participant. On days 1, 2, 3, 4, and 5 the test run was directly followed by a self-assessment. In addition, on days 1, 3, and 5 the test run for each beginner was recorded on video, with each video assigned an encrypted code number. All the end of the study week, control assessments of these videos were carried out by an experienced endoscopist. RESULTS: Both the self-assessments and the control assessments showed significant improvements in the endoscopic parameters tested during the course (days 1-5; all parameters P < 0.001, Wilcoxon test). However, it was found that the trainees tended to give themselves better marks than the marks given by experienced endoscopists. CONCLUSION: During 1 week of training, using the model and following the score card, a significant improvement in the learning curve was achieved in the beginners' group for the individual steps involved in diagnostic upper gastrointestinal endoscopy. When this approach is used with trainees who are also provided with the necessary theoretical background, this type of preparation may lead to a better, lower-risk start to supervised practical endoscopic examinations in patients. PMID- 12783352 TI - Nonspecific ulcers of the colon. AB - Nonspecific colonic ulcers (NSCUs) are rare and potentially life-threatening lesions of unknown etiology; the diagnosis is based on histologic findings showing nonspecific inflammatory changes. The condition's variable symptoms can include nonspecific abdominal discomfort, gastrointestinal bleeding, perforation, and peritonitis. Radiologic imaging can be helpful in locating the lesions, but colonoscopy facilitates early definitive diagnosis and aggressive treatment. The potential of NSCUs to recur is currently unknown, and morbidity rates remain high. Long-term colonoscopic follow-up may improve the prognosis. PMID- 12783353 TI - Pneumatic dilation in achalasia. PMID- 12783354 TI - Hemobilia in hereditary hemorrhagic telangiectasia: an unusual complication of endoscopic retrograde cholangiopancreatography. AB - Hepatobiliary manifestations of hereditary hemorrhagic telangiectasia (HHT) are rare, but often involve cholestasis. We report here a case of HHT associated with cholestasis due to common bile duct stenosis. Attempted balloon dilation of the stenosis during endoscopic retrograde cholangiopancreatography (ERCP) resulted in hemobilia. Hemostasis was achieved by adjusting the nasobiliary drain. The aim of this report is to highlight the biliary manifestations of HHT and draw attention to an unusual complication of ERCP in this setting. PMID- 12783355 TI - Pancreatic intraductal ultrasonography (IDUS) allows early diagnosis of pancreatic carcinoma in situ: a case report. AB - A 66-year-old woman was admitted with diarrhea, weight loss, slight recurrent abdominal pain, and raised serum amylase and lipase. Lactose intolerance was diagnosed, and treatment was begun. The symptoms diminished. However, slight back pain and elevated serum amylase and lipase levels persisted. A pancreatic tumor was then suspected. Ultrasound, spiral computed tomography (CT), magnetic resonance imaging (MRI), and magnetic resonance cholangiopancreatography (MRCP) examinations were inconclusive. Endoscopic retrograde cholangiopancreatography (ERCP) showed a slight narrowing of the pancreatic duct within the pancreatic body, and endoscopic ultrasound (EUS) revealed a 10 mm intrapancreatic lesion. Finally, intraductal ultrasonography (IDUS) reliably identified a small pancreatic tumor. The tumor was resected, and histology confirmed a well differentiated adenocarcinoma in situ (UICC stage 0, TisN0M0). This case shows that using high-resolution imaging techniques such as EUS plus IDUS, small malignant pancreatic lesions can be detected at an early stage, when curative action is possible. PMID- 12783356 TI - Early neoplasias of the gallbladder and bile duct: an "unstable" biliary epithelium? AB - Benign tumours of the biliary tree are rare. In particular, only anecdotal cases of intraductal villous adenomas have been reported. The polyp-cancer sequence has not been observed in the biliary epithelium, in contrast to the paradigm of colorectal carcinogenesis. This report presents the case of a 64-year-old woman with a past history of cholelithiasis who had two early neoplasias involving the biliary epithelium: an adenocarcinoma in situ of the gallbladder and a common bile duct (CBD) villous adenoma with high-grade dysplasia. The tumours presented 4 years apart. The clinical features and combined radiological, cytological, and surgical modalities leading to the diagnosis of intraductal villous adenoma are presented. The endoscopic ultrasound (EUS) characteristics of villous adenoma of the CBD are described. While the prognosis on both occasions appears excellent following curative resections of both tumours detected at an early stage, it is possible that further neoplasia involving the biliary tree may recur. There are currently no data on optimal surveillance modalities. It may be hypothesized that the gallbladder and biliary epithelium share a similar mechanism for carcinogenesis to that observed in the colonic adenomacarcinoma sequence. PMID- 12783357 TI - Factor V inhibitor after injection of human thrombin (tissucol) into a bleeding peptic ulcer. AB - Endoscopic injection of fibrin glue into a bleeding peptic ulcer is an effective and safe treatment modality. The present report describes a patient who developed rectal bleeding from an arteriovenous malformation after endoscopic injection of fibrin glue containing human thrombin into a gastric ulcer. Additional laboratory investigations revealed the presence of an inhibitor against coagulation factor V, which resulted in severe coagulopathy, triggering the bleeding. Acquired factor V inhibitors have frequently been reported with the use of bovine thrombin, but to our knowledge, they have never been documented in patients exposed to human thrombin. Endoscopists should be aware of this rare, but potentially serious, complication. PMID- 12783358 TI - Ectopic drainage of the common bile duct into the duodenal bulb: report of two cases. PMID- 12783359 TI - Intercostal positioning of a percutaneous endoscopic gastrostomy. PMID- 12783360 TI - Ectopic pancreas presenting as a large gastric antral papilla. PMID- 12783361 TI - Modified suction cap technique for endoscopic management of esophageal food impaction. PMID- 12783362 TI - Jejunoduodenal fistula due to penetrating peptic ulcer after Roux-en-Y hepaticojejunostomy. PMID- 12783363 TI - An unusual cause of colon polyps. PMID- 12783364 TI - Combination of colonoscopy and clip application with angiography to mark vascular malformation in the small intestine. PMID- 12783368 TI - Molecular mechanisms of transformation by the BCR-ABL oncogene. AB - The BCR-ABL oncogene is generated by the Philadelphia chromosome (Ph) translocation, fusing the BCR gene to the ABL gene. The BCR-ABL fusion protein has elevated ABL tyrosine kinase activity that is critical for transformation of hematopoietic cells. Chronic myelogenous leukemia (CML) cells transformed by BCR ABL show reduced growth factor requirements and apoptosis, as well as enhanced viability and altered adhesion. The elevated ABL kinase activity leads to chronic activation of signaling pathways that are required for all aspects of transformation. Progression of the disease from chronic phase to blast crisis correlates with additional cytogenetic alterations that are likely to contribute to the failure of traditional therapy. This review describes molecular mechanisms that are thought to be important for transformation by the BCR-ABL oncoprotein and points at pathways for targeted drug development in the treatment of CML. PMID- 12783367 TI - Chronic myeloid leukemia in the imatinib era. AB - The results of imatinib (Gleevec) (formerly STI571) therapy for chronic myeloid leukemia (CML) have continued to improve and have surpassed almost everyone's predictions. However, the long-term results with imatinib and its ability to cure CML as a single agent are unknown. These issues will be discussed along with potential strategies to improve the cure rate, using imatinib as a building block to achieve this goal. PMID- 12783369 TI - Towards combination target-directed chemotherapy for chronic myeloid leukemia: role of farnesyl transferase inhibitors. AB - Chronic myeloid leukemia (CML) is arguably the best understood of all human malignancies. Its origins in the hematopoietic stem cell can be traced to a reciprocal translocation involving chromosomes 9 and 22, dubbed the Philadelphia chromosome, which is observed in essentially all patients. The resulting fusion gene, BCR/ABL, encodes an activated tyrosine kinase that can act alone to induce a CML-like syndrome in mouse models. These animal models have validated BCR/ABL as a target for the development of specific pharmaceutical inhibitors. The kinase inhibitor imatinib mesylate (Gleevec) is highly specific, effective, and minimally toxic, but may not effect cures as a single agent, particularly in patients with accelerated and blast-phase disease. Resistance to imatinib can confound therapy. Surprisingly, a high percentage of resistant cases manifest intact or augmented BCR/ABL signaling, suggesting that this oncoprotein, or signaling pathways emanating from it, remain viable targets. Combination chemotherapy is under active investigation, and among the most compelling strategies is dual treatment with agents that both target BCR/ABL signal transduction. BCR/ABL activates Ras, and compounds designed to antagonize Ras function called farnesyl transferase inhibitors (FTIs) have shown potent activity in vitro and in animal models of BCR/ABL-induced leukemia. Initial clinical trials in patients with refractory acute myeloid leukemia and CML in blast crisis have shown significant activity, suggesting that trials combining imatinib and FTIs are warranted. PMID- 12783370 TI - Imatinib: a targeted clinical drug development. AB - Imatinib (Gleevec) (formerly STI571) is an orally bioavailable rationally developed inhibitor of the tyrosine kinases Bcr-Abl, Kit, and platelet-derived growth factor receptor (PDGFR). In 4 years of clinical development, more than 12,000 patients have been treated in the clinical development program. Imatinib was first shown to be highly effective in the treatment of all stages of chronic myelogenous leukemia (CML). Moreover, preliminary results of a randomized study have demonstrated superior efficacy and safety of first-line imatinib therapy compared with a combination of interferon and cytarabine. Imatinib has also been shown to be the only effective drug therapy in the treatment of patients with metastatic gastrointestinal stromal tumors expressing the stem cell factor (SCF) receptor Kit. This review outlines the successive steps in the clinical development of this new, targeted anticancer agent. PMID- 12783371 TI - Imatinib treatment: specific issues related to safety, fertility, and pregnancy. AB - Imatinib (Gleevec) (formerly STI571) has demonstrated high levels of efficacy in chronic myelogenous leukemia (CML) and gastrointestinal stromal tumors (GIST) and has been used in more than 12,000 patients participating in clinical trials. Experience from clinical trials with imatinib has largely demonstrated the drug to be well tolerated in humans. Common side effects, usually manageable, include nausea, rash, superficial edema, myelosuppression, muscle cramps, and elevated liver transaminases. With longer follow-up and with further experience with the treatment of patients outside of clinical trials, we are able to report on rarer toxicities, the identification of certain predictors of common toxicities, and the clinical experience with male fertility and pregnancy outcomes. PMID- 12783372 TI - Imatinib in patients with newly diagnosed chronic-phase chronic myeloid leukemia. AB - The International Randomized Study of Interferon and STI571 (IRIS) study prospectively compared imatinib with interferon-alpha/low-dose cytarabine (IFN/LDAC) in 1,106 newly diagnosed patients with Philadelphia chromosome positive chronic myeloid leukemia (CML). Patients not responding to or intolerant of their assigned treatment were allowed to cross over. At 18 months, the projected probability of achieving a complete cytogenetic response was 76.2% for imatinib and 14.5% for IFN/LDAC, respectively (P <.01). Freedom from progression to accelerated phase or blast crisis was 96.7% for imatinib versus 91.5% for IFN/LDAC (P <.01). At the time of the analysis, 85.7% of imatinib-treated patients continued on first-line therapy, but only 10.8% of patients continued with IFN/LDAC. Most cross-overs to imatinib were due to interferon-intolerance. Overall survival was not different in the two groups at 19 months, reflecting efficient rescue of IFN/LDAC failures with imatinib. Imatinib should now be considered the standard therapy for newly diagnosed patients with CML. PMID- 12783373 TI - Quality of life on imatinib. AB - Imatinib (Gleevec), a highly effective specific tyrosine kinase inhibitor, demonstrates a better side effect profile than interferon-alpha (IFN), which impairs patients' quality of life (QoL). This phase III international study evaluated QoL outcomes in 1,106 newly diagnosed patients with chronic-phase chronic myeloid leukemia (CML) who were randomized to receive either imatinib 400 mg daily or IFN up to 5 MU/m(2)/d with cytarabine (Ara-C) 20 mg/m(2)/d added for 10 days every month (IFN + LDAC). Crossover to the other treatment arm was permitted due to a lack of efficacy or treatment intolerance. QoL was assessed with the Functional Assessment of Cancer Therapy-Biologic Response Modifiers (FACT-BRM) at baseline, monthly for 6 months and then at months 9, 12, and 18. The Trial Outcome Index (TOI; a composite endpoint of physical/functional/treatment-specific subscales) was the primary endpoint. Secondary endpoints measured were social/family well-being (SFWB) and emotional well-being (EWB). QoL was analyzed for the first 18 months of treatment using mixed effects growth curve models. The primary analyses were intention-to-treat (ITT); secondary analyses incorporated crossover as a time-dependent covariate. A total of 1,049 patients completed at least one QoL assessment. Two hundred sixty one patients (50%) crossed over from IFN to imatinib and 11 (2%) crossed over from imatinib to IFN. There was a significant decline in TOI scores for the IFN treatment arm compared with preservation of baseline TOI scores in the imatinib arm (P <.001, ITT). Mean social/family and EWB scores were 22.8 and 19.5, respectively, for imatinib and 21.6 and 17.6, respectively, for IFN (P <.001, ITT). After crossing over from IFN to imatinib, patients experienced a significant (P <.001) increase in TOI scores. Imatinib offers clear QoL advantages over IFN as first-line treatment of chronic-phase CML. In addition, patients who crossed over to imatinib reported higher QoL than those who remained on IFN. Semin Hematol 40(suppl 2):31-36. PMID- 12783374 TI - Usefulness of quantitative assessment of Wilms tumor suppressor gene expression in chronic myeloid leukemia patients undergoing imatinib therapy. AB - The Wilms tumor suppressor gene (WT1) is overexpressed in a number of human hematological malignancies, including chronic myeloid leukemia (CML). Although at present, the biological significance of WT1 expression in CML in still unclear, this marker could represent a useful tool for molecular monitoring of CML patients prior to and post imatinib therapy. In fact, the use of real-time polymerase chaine reaction (PCR) to quantitatively measure the WT1 transcript amount may be a predictor of patient response to imatinib therapy. PMID- 12783375 TI - Prediction of response to imatinib by cDNA microarray analysis. AB - Clinical introduction of imatinib mesylate (Gleevec) (formerly STI571) has marked a major advance in the treatment of chronic myeloid leukemia (CML), inducing major cytogenetic response in 60% to 80% of patients with chronic-phase disease. Since its remarkable effect has made it difficult to decide the timing for potentially curative but life-threatening stem cell transplantation (SCT), it would be very useful to predict imatinib's effectiveness in individual patients beforehand. We analyzed gene expression profiles of peripheral leukemia cells from 18 CML patients (16 chronic phase, two blast crisis) who were treated with imatinib alone. cDNA microarrays representing 23,040 genes identified 79 genes that were expressed differentially between cytogenetically defined responders and nonresponders to imatinib. We then established a "prediction score" system using 15 or 30 of these genes that could clearly separate the responder group from the nonresponder group. Four additional patients' responses to imatinib were successfully predicted, validating the process. Thus, gene-expression profiles can predict clinical effectiveness of CML to imatinib, and may eventually lead to the achievement of "personalized therapy" for this disease. PMID- 12783376 TI - Cytogenetic studies in patients on imatinib. AB - With the introduction of imatinib (Gleevec) (formerly STI571) for the treatment of chronic myeloid leukemia (CML), the various diagnostic methods used to monitor patients must be re-evaluated. Conventional cytogenetics has been the established method for follow-up of patients treated with interferon-alpha (IFN-alpha), and the prognostic value of major and complete cytogenetic remission was demonstrated in a number of studies. In patients on imatinib, these endpoints will likely remain valid, although longer observation is required. Cytogenetic remission as a time-dependent variable may aid risk stratification early on. Clonal evolution, ie, the presence of cytogenetic abnormalities in addition to the Philadelphia (Ph) chromosome, may provide important prognostic information as to the likely response to imatinib in all phases of CML but must be interpreted within the context of other disease characteristics. In some patients, clonal evolution is related to imatinib resistance. Information regarding the impact of specific types of additional cytogenetic abnormalities is still limited. Surprisingly, nonrandom karyotypic abnormalities have also been noted in the Ph-negative cells of some patients in cytogenetic remission. This is a novel phenomenon whose causality and prognostic implications require thorough and systematic evaluation. PMID- 12783377 TI - The cytogenetic response as a surrogate marker of survival. AB - In the management of chronic myeloid leukemia (CML), cytogenetic response (CgR) has been recognized as an important prognostic factor and is used in predicting patient outcome. Imatinib (Gleevec) (formerly STI571), a potent inhibitor of BCR ABL, is very effective in inducing CgRs in chronic-phase CML patients, even in late chronic-phase patients in whom interferon (IFN) was unsuccessful. This review describes the intrinsic value of a CgR following different forms of nontransplant treatment options for CML. Achievement of major CgR is predictive of prolonged survival with imatinib. In addition, patient prognostic risk score (Sokal or Euro risk score) affects patient outcome. Low-risk patients survive longer than non-low-risk patients given the same CgR. Preliminary results from studies with imatinib are superior and CgR remains an important surrogate marker of survival in the imatinib era. PMID- 12783378 TI - Molecular monitoring of chronic myeloid leukemia. AB - The tyrosine kinase inhibitor imatinib mesylate (Gleevec) (formerly STI571) has proven to be an effective and safe new therapy for patients with chronic myeloid leukemia (CML). It has induced short-term hematologic control in many patients with advanced-phase CML, with some patients achieving durable responses. In chronic-phase patients it induces significantly better cytogenetic responses and lower progression rates than interferon-alpha. However, relapse is a significant problem, especially for advanced-phase patients, and imatinib alone appears unlikely to be curative in any patient group. Real-time quantitative polymerase chain reaction (Q-PCR) provides an accurate, sensitive, and noninvasive measure of residual leukemia in patients on imatinib. Levels of BCR-ABL in the blood correlate strongly with the bone marrow cytogenetic results and early measurement can predict subsequent cytogenetic response. Complete molecular responses (no BCR ABL detected by real-time Q-PCR) are rarely achieved. Sequential real-time Q-PCR studies should facilitate rational patient management and allow comparison of different imatinib-based treatment strategies. It may be possible to define levels of molecular response that predict long-term disease control. In addition, by defining patterns of response, an early indication of imatinib resistance may be detected. PMID- 12783379 TI - Cytogenetic and molecular mechanisms of resistance to imatinib. AB - Selective inhibition of the BCR-ABL tyrosine kinase by imatinib (Gleevec) (formerly STI571) is a promising new therapeutic strategy in patients with chronic myelogenous leukemia (CML). Despite significant hematologic and cytogenetic responses, resistance occurs in patients with chronic phase (CP) and advanced disease. A cohort of 72 patients with CML in myeloid blast crisis (BC) (n = 34), lymphoid BC (n = 2), accelerated phase (AP) (n = 16), CP (n = 18), and BCR-ABL(+) acute lymphoblastic leukemia (ALL) (n = 2) resistant to imatinib were investigated. Median levels of BCR-ABL transcripts, determined by quantitative reverse-transcriptase polymerase chain reaction (RT-PCR), were not significantly changed at the time of resistance, but seven of 55 patients showed a greater than 10-fold increase in BCR-ABL levels. Genomic amplification of BCR-ABL was found in two of 32 patients evaluated by fluorescence in situ hybridization (FISH). Additional chromosomal aberrations were observed in 19 of 36 patients and point mutations of the ABL tyrosine kinase domain resulting in reactivation of the BCR ABL tyrosine kinase were detected in 29 of 72 patients. Resistance may be caused by BCR-ABL-independent or BCR-ABL-dependent mechanisms. A thorough evaluation of resistant cases is required to suggest therapeutic measures in the individual case. Clonal selection of resistant cells harboring a BCR-ABL mutation might be reversed by stopping imatinib therapy and switching to chemotherapy. Combination therapy from the start of treatment to reduce the frequency of resistance is currently being evaluated with several drugs. PMID- 12783381 TI - Imatinib mesylate in combination with other chemotherapeutic drugs: in vitro studies. AB - Imatinib mesylate (Gleevec) (formerly STI571) has secured a definitive role in the treatment of chronic myeloid leukemia (CML) due to its specificity and efficacy. Although some patients become resistant to the drug, it may still be possible to control the leukemia with imatinib-containing regimens. Front-line treatment with such combinations may indeed minimize the risk that resistance, and hence relapse, occurs. In this review, we discuss the published data on in vitro studies that address this question in a variety of models, and attempt to predict efficacious combinations for future clinical trials. These represent regimens where imatinib is combined with conventional chemotherapeutic drugs or with inhibitors of other key signal transduction molecules that may be preferentially activated in CML cells. PMID- 12783380 TI - Mutations in the ABL kinase domain pre-exist the onset of imatinib treatment. AB - Imatinib (Gleevec) (formerly STI571) competitively targets the adenosine 5 triphosphate (ATP) binding site of the kinase domain of ABL and was recently approved for the treatment of chronic myeloid leukemia (CML). Point mutations occurring in the kinase domain of BCR-ABL have been identified as a cause of imatinib resistance. These mutations can be categorized into two groups: (1) mutations directly impairing the binding of imatinib but not ATP, and (2) mutations occurring in the ATP phosphate binding loop (P loop) or activation loop preventing the kinase to achieve conformation required for imatinib binding. Functional analysis of mutant BCR-ABL alleles in vitro has demonstrated four mutations (Q252H, F317L,M351T, E355G) to confer moderate resistance to imatinib, while T315I-, E255K-, Y253F-, and G250E-expressing cells are markedly resistant. Assay sensitivities and patient selection will affect the frequency of mutation detection. Another possible explanation for imatinib resistance is that mutated BCR-ABL-expressing cells might pre-exist the onset of treatment at levels below threshold detection (<20%), then expand under selective pressure of imatinib treatment. Rare mutated cells were identified using a very sensitive allele specific oligonucleotide polymerase chain reaction (ASO-PCR) assay in pretreatment samples of five CML patients supporting the theory that point mutations pre-existed imatinib treatment. Imatinib-resistant patients may benefit from molecular genotyping so mutations can be identified and clinical therapy adjusted accordingly. PMID- 12783382 TI - Imatinib in combination with cytarabine for the treatment of Philadelphia positive chronic myelogenous leukemia chronic-phase patients: rationale and design of phase I/II trials. AB - Imatinib (Gleevec) (formerly STI571) has been shown to selectively inhibit the tyrosine kinase domain of the oncogenic BCR-ABL fusion protein of chronic myelogenous leukemia (CML) cells. In recent phase I and II studies testing this new compound in patients who had failed to respond to interferon (IFN), hematological and cytogenetic responses were reported in most of those with chronic-phase CML. However, in some patients resistance has been associated with a single amino acid substitution in a threonine residue of the Abl kinase domain. In vitro studies examining the effects of imatinib plus cytarabine (Ara-C) using CML cell lines and colony-forming assays of CML patient samples have shown synergistic antiproliferative effects of this combination. Thus several groups decided to investigate this new combination with the hypothesis that cell resistance would be less frequent. The CML French Group performed a phase II trial to determine the safety and tolerability of a combination of imatinib and Ara-C for previously untreated patients with chronic-phase CML. Treatment was administered on 28-day cycles for 12 months. Patients were treated continuously with imatinib orally at a dose of 400 mg daily. Ara-C was given on days 14 to 28 of each cycle at an initial dose of 20 mg/m(2)/d via subcutaneous injection, hydroxyurea (HU) being stopped at least 7 days before imatinib. Recently, the Dutch group decided to explore a combination of high-dose Ara-C with imatinib in patients in chronic-phase CML. Preliminary results are encouraging. However, a long follow-up is required before concluding that these strategies will overcome cell resistance. PMID- 12783383 TI - Clinical decisions for chronic myeloid leukemia in the imatinib era. AB - Imatinib is clearly an important contribution to the management of patients with chronic myeloid leukemia (CML). Based on the available data, it seems reasonable to conclude that this exciting new agent is the best primary treatment for the newly diagnosed chronic-phase patient who is not immediately eligible for allogeneic stem cell transplant (allo-SCT). However, the definition of response or failure to respond is not straightforward and the extent to which imatinib prolongs life when used as a single agent cannot yet be estimated with any accuracy. This means that decisions on how best to continue treatment can be extremely difficult. Moreover, the decision whether and when to offer the patient treatment by allo-SCT has become especially complicated in the last 2 years. Imatinib seems to be generally less effective when used to treat patients in advanced phases of CML, although some patients treated in accelerated phase may achieve prolonged disease control. It is likely that the role of imatinib, used alone or in combination with other agents, will be more clearly defined in the near future. PMID- 12783392 TI - Recurrent primary biliary cirrhosis. AB - Liver transplantation remains the only effective treatment for end-stage primary biliary cirrhosis (PBC). It appears now well accepted that the disease recurs in the allograft. The diagnosis of recurrent PBC is made on the basis of a consistent history and demonstrating the histologic features of PBC on liver biopsy and exclusion of other causes of bile duct damage. PMID- 12783393 TI - Relationship between portal venous flow and liver regeneration in patients after living donor right-lobe liver transplantation. AB - The purpose of this study is to evaluate the relationship between portal venous (PV) velocity and degree of liver regeneration in humans after living donor liver transplantation (LDLT). Between July 1997 and September 2002, a total of 15 adult to-adult LDLTs with right-lobe grafts were performed, and 13 of these patients were enrolled in this study. Postoperative PV dynamics differed according to the primary liver disease; therefore, patients were divided into two groups: a fulminant hepatic failure (FHF) group (n = 4) and a liver cirrhosis (LC) group (n = 9). Right-lobe donors (n = 13; D group) were used as controls. Doppler ultrasound was used to measured changes in PV velocity preoperatively; postoperative days (PODs) 1, 3, 7, 14, and 28; and 3 months after LDLT. To assess hepatic regeneration, the increase in liver volume ratio (postoperative liver volume to standard liver volume [SLV]) was measured. PV velocity after LDLT in the LC group increased sharply until POD 7, whereas those in the FHF and D groups were constant. In the first 3 months after LDLT, mean PV velocity was greater in the LC group than the other groups, reflecting the persistent hyperdynamic state in chronic end-stage liver disease. Liver regeneration also was more rapid in the LC group than the FHF and D groups and reached 100% as early as 2 weeks posttransplantation, whereas both the FHF and D livers reached approximately 80% of SLV at 3 months. PV velocity POD 7 correlated significantly with regeneration of the partial-liver allograft at 1 month (r = 0.84; P =.0091). In conclusion, the PV persistent hyperdynamic state in the LC group could directly trigger early liver regeneration in partial-liver allografts after LDLT. PMID- 12783394 TI - Extensive chimerism in liver transplants: vascular endothelium, bile duct epithelium, and hepatocytes. AB - The transplanted liver has been shown to be particularly capable of inducing tolerance. An explanation may be the presence of chimerism. Cells of donor origin have been found in recipient tissues after transplantation of any solid organ. Evidence for the presence of cells of recipient origin within the transplanted liver is very limited. We investigated whether nonlymphoid cells of recipient origin can be found within human liver allografts. Five male patients who received a liver transplant from a female donor and 11 patients who received an HLA-I mismatched liver transplant were studied. We confirmed our observations with two different techniques in combination with double-staining techniques. To identify male cells in female liver transplants, we used in situ hybridization for sex chromosomes. To identify specific HLA class I antigens of recipient origin, we used immunohistochemistry with HLA class I-specific antibodies. Double staining was performed to discriminate different cell lineages and inflammatory cells. Endothelial cells of recipient origin were found in 14 of 16 donor livers. Bile duct epithelial cells of recipient origin were found in 5 of 16 cases. Hepatocytes of recipient origin were seen in only 1 of the 5 studied sex mismatched donor livers. Our study provides evidence that cells of recipient origin can replace biliary epithelial cells, endothelial cells, and hepatocytes within the human liver allograft. This is consistent with the concept that circulating pluripotent progenitor cells exist, capable of differentiating into endothelial cells, epithelial cells, and hepatocytes. PMID- 12783396 TI - Changes in portal vein flow after adult living-donor liver transplantation: does it influence postoperative liver function? AB - In adult living donor liver transplantation, using small grafts in cirrhotic patients with severe portal hypertension may have unpredictable consequences. The so-called small-for-size syndrome is present in most series worldwide. The goal of this study was to prospectively evaluate the influence of hemodynamic changes on postoperative liver function and on the percentage of liver volume increase, in the setting of living donor liver transplantation. Twenty-two consecutive adult living donor liver transplantations were performed at our institution in a 2-year period. We measured right portal flow and right hepatic arterial flow with an ultrasonic flow meter in the donor, and then in the recipient 1 hour after reperfusion. Postoperative liver function was measured by daily laboratory work. We also performed duplex ultrasounds on postoperative days 1, 2, and 7. Liver volume increase was estimated by magnetic resonance imaging graft volumetry at 2 months posttransplantation. We compared the blood flow results with the immediate liver function and its liver volume increase rate at 2 months. There was a significant increase in portal flow in the recipients compared with the donors (up to fourfold in some cases). Higher portal flow increase rates significantly correlated with faster prothrombin time normalization and faster liver volume increases. Median graft volume increase at 2 months was 44.9%. The increase in blood flow to the graft is well tolerated by the liver mass not affecting hepatocellular function as long as the graft-to body weight ratio is maintained (>0.8) and adequate outflow is provided. PMID- 12783395 TI - Chemoembolization followed by liver transplantation for hepatocellular carcinoma impedes tumor progression while on the waiting list and leads to excellent outcome. AB - Orthotopic liver transplantation (OLT) has been considered the best treatment option for patients with hepatocellular carcinoma (HCC). Because of a steadily increasing waiting time, a noteworthy proportion of patients are excluded from OLT because of tumor progression. A 20% and more dropout rate from the waiting list has recently been reported. In this prospective study, we evaluated the effect of preoperative transarterial chemoembolization (TACE) on preventing tumor progression while on the waiting list in patients meeting current selection criteria (solitary lesion < or = 5 cm, three lesions < or = 3 cm). In addition, we analyzed the outcome of a separate group of patients with advanced-stage HCC outside the selection criteria but with at least 50% tumor reduction after TACE (downstaging) to expand current criteria. Forty-eight patients met the selection criteria and were eligible for this study. Seven patients are still on the waiting list; 41 underwent OLT. None of these patients had to be removed from the list because of tumor progression after a mean waiting time of 178 days (23 patients > or =180 days). The 1-, 2-, and 5-year intention-to-treat survival was 98%, 98%, and 94%. The outcome after OLT was also excellent with 1-, 2-, and 5 year survival rates of 98%, 98%, and 93%. Tumor recurrence occurred only in 1 patient (2.4%). Fifteen patients with advanced-stage HCC were included in this study. Three developed a tumor progression and had to be removed from the list (20% dropout rate). Despite tumor reduction before OLT, these patients had a significantly less favorable outcome in the intention-to-treat analysis as well as in the posttransplantation survival. Tumor recurrence was seen in 30% of patients after OLT. In conclusion, TACE followed by OLT is associated with an excellent outcome in selected patients. Furthermore, TACE is highly efficacious in preventing tumor progression while waiting for OLT. Although TACE reduced tumor preoperatively, it failed to show a beneficial effect on patient survival in advanced-stage HCCs. PMID- 12783397 TI - Hepatic arterial reconstruction in 95 adult right lobe living donor liver transplants: evolution of anastomotic technique. AB - The success of adult right-lobe living donor liver transplants (RLDLT) would not have been possible without the experience of the early pioneers of pediatric living donor liver transplants (LDLT). Our experience with 95 RLDLTs from July, 2000 to May, 2002 at a single institution is reported with specific emphasis on arterial reconstruction. The evolution of technique using the autogenous Y extension graft and the reverse extension bifurcated graft for arterial revascularization, which we believe has reduced the incidence of hepatic artery thrombosis, is described. Outcome and the incidence of hepatic artery thrombosis are reported. PMID- 12783398 TI - Biliary reconstructions and complications encountered in 50 consecutive right lobe living donor liver transplantations. AB - Biliary complications appear to be the leading cause of postoperative complications after living donor liver transplantation (LDLT). The aim of this study is to analyze the complications, treatment modalities, and outcomes of biliary anastomoses in a series of 50 consecutive right-lobe LDLTs. Median patient age was 45 years, and median right-lobe graft volume was 740 g. Graft recipient weight ratio was 0.69 to 1.80. Median follow-up time was 15 months (range, 2 to 38 months). Eleven of 50 patients died, resulting in an overall allograft and patient survival rate of 78%. In biliary reconstruction, a duct-to duct (D-D) anastomosis or a standard Roux-en-Y (R-Y) anastomosis was performed. Twenty-nine grafts (58%) had a single duct for anastomosis. Seventeen grafts (34%) had two bile duct orifices, and four grafts (8%) had three bile duct orifices. A D-D anastomosis was performed in 36 cases (72%), whereas R-Y reconstruction was preferred in 14 cases (28%). The overall incidence of biliary anastomotic complications was 30% in this series. Five patients developed biliary leaks, presumably from the cut surface, and all of them healed spontaneously. Two bilomas were drained percutaneously. Anastomotic strictures occurred in 8 patients (16%) and were significantly greater than in the R-Y group (P =.03). Although strictures seemed to develop more frequently in allografts with multiple bile ducts, this did not reach statistical significance (P =.05). All strictures were managed by nonsurgical measures initially. Restenosis occurred in 2 patients, both of whom had an R-Y anastomotic stricture. These anastomoses were revised surgically, giving a reoperation rate of 4% for biliary problems. No graft or patient was lost because of biliary problems. Our data suggest that D-D anastomosis is a safe and feasible method of biliary reconstruction in LDLT by preserving physiological bilioenteric continuity and allowing easy access through endoscopic techniques. PMID- 12783399 TI - Outcome analysis in adult-to-adult living donor liver transplantation using the left lobe. AB - Graft size problems remain the greatest limiting factor for expansion of living donor liver transplantation (LDLT) to the adult population. The result of adult to-adult LDLT using the left lobe with special reference to graft size has not been fully evaluated to date. In this study, we evaluated the outcome of adult-to adult LDLT using the left lobe and also analyze the impact of using small-for size grafts on outcome. Thirty-six recipients who underwent adult-to-adult LDLT using the left lobe (n = 14) or left lobe plus caudate lobe (n = 22) were included in the study. Variables including preoperative and operative data, patient and graft survival, complications, and causes of graft loss were studied. Furthermore, the incidence of small-for-size syndrome and its impact on graft survival were studied. Mean graft volume (GV) was 420 +/- 85 g (range, 260 to 620 g), which resulted in 38.2% +/- 8.1% (range, 22.8% to 53.8%) of the recipient standard liver volume (SLV). Overall 1-year patient and graft survival rates were 85.7% and 82.9%, respectively. Seven grafts were lost. Small-for-size syndrome occurred in 7 of 16 patients (43.8%) with cirrhosis and only 1 of 20 patients (5.0%) without cirrhosis (P =.005). Recipients who developed small-for-size syndrome had inferior graft survival to those who did not (P =.07). In conclusion, adult-to-adult LDLTs were found to be feasible without affecting patient or graft survival. Small-for-size syndrome developed more frequently in patients with cirrhosis. Minimum GV in adult-to-adult LDLT should be 30% less than the recipient's SLV in patients without cirrhosis, whereas 45% less was required in patients with cirrhosis. PMID- 12783400 TI - Early steroid withdrawal after liver transplantation: the Canadian tacrolimus versus microemulsion cyclosporin A trial: 1-year follow-up. AB - Corticosteroid therapy contributes significant toxicity to liver transplantation. The safety and efficacy of early steroid withdrawal were determined in patients treated with either tacrolimus or microemulsion cyclosporin A (micro-CsA). The primary outcome was the proportion of patients who were steroid-free 1 year posttransplantation. From the seven Canadian adult liver transplant centers, 143 patients were randomly allocated oral treatment with either tacrolimus (n = 71) or micro-CsA (n = 72), together with corticosteroids and azathioprine. Eligibility criteria for steroid withdrawal included freedom from acute rejection for a minimum of 3 months, and prednisone 80-fold). Immunohistochemistry showed MMP-12 to be localized in activated monocytoid cells surrounding the hematoma. In vitro studies showed that thrombin, released during ICH, induced MMP-12 expression in monocytoid cells, which was reduced by minocycline application. Similarly, in vivo minocycline treatment significantly reduced MMP-12 levels in brain. Neuropathological studies disclosed marked glial activation and apoptosis after ICH that was reduced by minocycline treatment. Neurobehavioral outcomes also were improved with minocycline treatment compared with untreated ICH controls. Thus, select MMPs exhibit increased expression after ICH, whereas minocycline is neuroprotective after ICH by suppressing monocytoid cell activation and downregulating MMP-12 expression. PMID- 12783420 TI - Statins induce angiogenesis, neurogenesis, and synaptogenesis after stroke. AB - We demonstrate that the 3-hydroxy-3-methyl-glutaryl-coenzyme A (HMG-CoA) reductase inhibitors atorvastatin and simvastatin enhance functional outcome and induce brain plasticity when administered after stroke to rats. With atorvastatin treatment initiated 1 day after stroke, animals exhibited significant increases in vascular endothelial growth factor, cyclic guanosine monophosphate, angiogenesis, endogenous cell proliferation and neurogenesis, and an increase in the synaptic protein, synaptophysin. Atorvastatin-induced angiogenesis in a tube formation assay was reduced by an antibody against the vascular endothelial growth factor receptor 2 (FIK-1) and by the nitric oxide synthase inhibitor, N mono-methyl-L-arginine (L-NAME). Atorvastatin also induced phosphorylation of Akt and Erk in cultured primary cortical neurons. These data indicate that atorvastatin induced brain plasticity and has neurorestorative activity after experimental stroke. PMID- 12783423 TI - Cortical anomalies associated with visuospatial processing deficits. AB - Children born preterm provide a fruitful population for studying structure function relationships because they often have specific functional deficits in the context of normal neurological status. We selected a group of preterm adolescents with deficits in judgment of line orientation. Despite their very low birth weight, all were neurologically normal with no consistent abnormalities on conventional magnetic resonance imaging. However, voxel-based morphometric analysis of their magnetic resonance imaging scans showed areas of decreased gray matter and increased white matter most prominently in right ventral extrastriate cortex, close to an area previously implicated in the line orientation task. We suggest that these anomalies of cortical architecture relate to impaired performance on the line orientation task. PMID- 12783421 TI - The spectrum of autoimmune autonomic neuropathies. AB - We analyzed the clinical characteristics of 18 patients (13 female, 5 male) who had autoimmune autonomic neuropathy (AAN) and ganglionic acetylcholine receptor (AChR) autoantibodies. Mean age was 61.4 years (standard deviation, 12.0 years). Ten patients had subacute symptom onset, six with an antecedent event. Eight patients had chronic AAN, characterized by insidious symptom onset, without antecedent event, and gradual progression. A majority of patients with high antibody values (>1.00 nmol/L) had a combination of sicca complex (marked dry eyes and dry mouth), abnormal pupillary light response, upper gastrointestinal symptoms, and neurogenic bladder. Chronic AAN segregated into two subgroups. One subgroup (N = 4) had low antibody titer (0.09 +/- 0.01 nmol/L) and a paucity of cholinergic symptoms. It was indistinguishable from pure autonomic failure. The other subgroup (N = 4) had high antibody titer (11.6 +/- 2.08 nmol/L), sicca complex, abnormal pupils, and neurogenic bladder; three had severe upper gastrointestinal dysfunction. Higher antibody titers correlated with greater autonomic dysfunction and more frequent cholinergic dysautonomia. These observations expand the clinical spectrum of AAN to include chronic cases, some being indistinguishable from pure autonomic failure, and support the concept that ganglionic AChR antibodies are important diagnostically and pathophysiologically in acquired dysautonomia. PMID- 12783422 TI - Functional organization of the brain with malformations of cortical development. AB - We examined the localization of cerebral functions in 28 patients with focal epilepsy and malformations of cortical development (MCDs). Polymicrogyria occurred in nine, hemimegalencephaly in four, heterotopia in eight, and focal cortical dysplasia (FCD) in nine cases. We used simple (sensomotor, visual) or complex (language, memory) functional magnetic resonance imaging (fMRI) paradigms. Two thirds of MCDs were activated by simple fMRI paradigms, whereas they less frequently showed activity during complex cognitive fMRI paradigms. During simple paradigms, all disturbances of cortical organization (polymicrogyria, schizencephaly, and mild-type FCD) showed activity, whereas other MCDs (disturbances of earlier steps of cortical development: hemimegalencephaly, Taylor-type FCD, and heterotopia) showed activity in only 44% (p < 0.01). The association between the pathophysiology and morphology of MCDs confirms the recently proposed classification system. Both focal neurological signs (p < 0.05) and focal electroencephalogram slowing (p < 0.05) independently correlated with MCD inactivity, confirming that fMRI showed neuronal functions of MCDs. Conclusively, fMRI visualizes the MCD functions and their relationship to the eloquent cortex, providing useful information before epilepsy surgery. Surgery of cortical organization disturbances should be cautiously performed because these malformations may participate to some degree in brain functions. PMID- 12783424 TI - Slowing of electroencephalogram in rapid eye movement sleep behavior disorder. AB - Rapid eye movement (REM) sleep behavior disorder (RBD) is characterized by a loss of atonia and an increase in phasic muscle activity during REM sleep, leading to complex nocturnal motor behaviors. Brainstem structures responsible for the pathogenesis of RBD are also implicated in cortical activation. To verify the hypothesis that electroencephalogram (EEG) activation will be impaired in RBD, we performed quantitative analyses of waking and REM sleep EEG in 15 idiopathic RBD patients and 15 age- and gender-matched healthy subjects. During wakefulness, RBD patients showed a considerably higher theta power in frontal, temporal, and occipital regions with a lower beta power in the occipital region. The dominant occipital frequency was significantly lower in RBD. During REM sleep, beta power in the occipital region was lower in RBD. This study shows for the first time an impaired cortical activation during both wakefulness and REM sleep in idiopathic RBD, despite an absence of changes on sleep architecture compared with controls. EEG slowing in these patients may represent an early sign of central nervous system dysfunction, perhaps paralleled by subclinical cognitive deficits. The topographical distribution of EEG slowing and possible pathophysiological mechanisms are discussed in light of the known association between RBD and neurodegenerative disorders. PMID- 12783425 TI - Toward therapy for DYT1 dystonia: allele-specific silencing of mutant TorsinA. AB - A three-nucleotide (GAG) deletion in the TOR1A gene is the most common cause of inherited dystonia, DYT1. Because the mutant protein, TorsinA (TA), is thought to act in a dominant manner to cause disease, inhibiting expression from the mutant gene represents a potentially powerful therapeutic strategy. In an effort to develop therapy for this disease, we tested whether small interfering RNA (siRNA) could selectively silence expression of mutant TA. Exploiting the three-base pair difference between wild-type and mutant alleles, we designed siRNAs to silence expression of mutant, wild-type, or both forms of TA. In transfected cells, siRNA successfully suppressed wild-type or mutant TA in an allele-specific manner: for example, mutant-specific siRNA reduced the levels of mutant TA to less than 1% of controls with minimal effect on wild-type TA expression. In cells expressing both alleles, thus simulating the heterozygous state, siRNA-mediated suppression remained robust and allele specific. Our siRNA studies demonstrate allele specific targeting of a dominant neurogenetic disease gene and suggest the broad therapeutic potential of siRNA for DYT1 dystonia and other dominantly inherited neurological diseases. PMID- 12783426 TI - Altered hippocampal muscarinic receptors in acetylcholinesterase-deficient mice. AB - A primary therapeutic strategy for Alzheimer's disease includes acetylcholinesterase (AChE) inhibitors with the goal of enhancing cholinergic transmission. Stimulation of muscarinic acetylcholine receptors (mAChRs) by elevated levels of ACh plays a role in the effects of AChE inhibitors on cognition and behavior. However, AChE inhibitors only demonstrate modest symptomatic improvements. Chronic treatment with these drugs may cause mAChR downregulation and consequently limit the treatment efficacy. AChE knockout (-/-) mice were utilized in this study as a model for investigating the effects of selective, complete, and chronic diminished AChE activity on mAChR expression and function. In AChE -/- mice, the M(1), M(2), and M(4) mAChRs showed strikingly 50 to 80% decreased expression in brain regions associated with memory. In addition, mAChRs showed decreased presynaptic, cell surface, and dendritic distributions and increased localization to intracellular puncta. Furthermore, mAChR agonist induced activation of extracellular signal-regulated kinase, a signaling pathway associated with synaptic plasticity and amyloidogenesis, is diminished in the hippocampus and cortex of AChE -/- mice. Therefore, chronic diminished ACh metabolism produces profound effects on mAChR expression and function. The alterations of mAChRs in AChE -/- mice suggest that mAChR downregulation may contribute to the limited efficacy of AChE inhibitors in Alzheimer's disease treatment. PMID- 12783427 TI - Effect of ultrasound on thrombolysis of middle cerebral artery occlusion. AB - We studied the effect of transcranial 2 MHz Doppler ultrasound (US) on the recanalization and outcome of stroke patients suffering from acute middle cerebral artery occlusion. All patients underwent intravenous recombinant tissue type plasminogen activator thrombolysis. Eleven patients were randomly selected for continuous US monitoring (C-US) over 1 hour; 14 patients were selected for the control group. The C-US group showed a higher grade of recanalization after 1 hour but also a higher number of intraparenchymal bleedings. Overall, a favorable functional outcome occurred more frequently in the C-US group (Barthel index, p = 0.037) after 3 months. PMID- 12783428 TI - Mitochondrial dysfunction in autistic patients with 15q inverted duplication. AB - Two autistic children with a chromosome 15q11-q13 inverted duplication are presented. Both had uneventful perinatal courses, normal electroencephalogram and magnetic resonance imaging scans, moderate motor delay, lethargy, severe hypotonia, and modest lactic acidosis. Both had muscle mitochondrial enzyme assays that showed a pronounced mitochondrial hyperproliferation and a partial respiratory chain block most parsimoniously placed at the level of complex III, suggesting candidate gene loci for autism within the critical region may affect pathways influencing mitochondrial function. PMID- 12783429 TI - Regular exercise is beneficial to a mouse model of amyotrophic lateral sclerosis. AB - We tested whether a regular exercise regimen was associated with a change in the life span of G93A-SOD1 transgenic mice, a model of familial ALS. Regular treadmill running for 10 weeks led to a significant increase in the life span of G93A-SOD1 mice. The effect was stronger in male mice, whereas there was only a trend between exercised and sedentary female G93A-SOD1 mice. The data suggest that regular exercise has a beneficial effect on the progression of ALS. PMID- 12783430 TI - Apolipoprotein C-II deficiency presenting as a lipid encephalopathy in infancy. AB - An infant presented with massive hyperchylomicronemia and a severe encephalopathy. MRI showed marked lipid deposition throughout the brain. Despite the normalization of the biochemistry, there was little clinical improvement, and at 18 months of age she has severe developmental delay, a strikingly abnormal MRI. Apolipoprotein C-II, the lipoprotein on chylomicrons responsible for the activation of lipoprotein lipase, was not detectable in blood. Analysis of the APO C-II gene revealed a novel homozygous point mutation, 1118C-->A. Subsequently, another sibling has been born with the same homozygous mutation and similar biochemistry but, perhaps because of early treatment, a normal neurological outcome. PMID- 12783431 TI - Healthy-side dominance of cortical neuromagnetic responses in sudden hearing loss. AB - Previous brain imaging and mapping studies have reported findings indicating functional reorganization in the central auditory pathways of patients with profound unilateral hearing loss. This study reports for the first time to our knowledge, using a whole-head neuromagnetometer with monaural stimulation of both intact and affected ears, a pattern of healthy-side dominance for cortical neuromagnetic responses in adult patients in the early stage of idiopathic sudden sensorineural hearing loss, and a pattern of contralateral dominance is verified in controls. PMID- 12783432 TI - N19S, a new SOD1 mutation in sporadic amyotrophic lateral sclerosis: no evidence for disease causation. AB - In a group of 331 sporadic amyotrophic lateral sclerosis (ALS) cases, we identified a new Cu/Zn superoxide dismutase single base substitution, N19S, in two patients. In the first case, seven healthy family members of 15 carried the substitution. Controls (n = 268) and familial ALS index cases (n = 180) were screened and one control subject with N19S was identified. Our data show that, despite a possible role of susceptibility factor for ALS, N19S alone cannot be considered as a direct cause for the disease. PMID- 12783433 TI - Elevated osteopontin levels in active relapsing-remitting multiple sclerosis. AB - In the search for proteins that might play a role in the pathogenesis of multiple sclerosis (MS), osteopontin (OPN) has been identified as the most prominent cytokine-encoding gene expressed within MS lesions. Here, we report significantly increased OPN protein levels in plasma of relapsing-remitting MS patients. In contrast, OPN protein levels in primary progressive and secondary progressive MS patients were similar to healthy control levels. Interestingly, active relapsing remitting patients had higher OPN protein levels than patients without relapses. PMID- 12783434 TI - Lack of association between the BDNF Val66Met polymorphism and Parkinson's disease in a Swedish population. PMID- 12783435 TI - Motor cortex hyperexcitability to transcranial magnetic stimulation in Alzheimer's disease: evidence of impaired glutamatergic neurotransmission? PMID- 12783437 TI - Influence of red wine on visual function and endothelin-1 plasma level in a patient with optic neuritis. PMID- 12783444 TI - Vertical agarose gel electrophoresis and electroblotting of high-molecular-weight proteins. AB - The electrophoretic separation of high-molecular-weight proteins (> 500 kDa) using polyacrylamide is difficult because gels with a large enough pore size for adequate protein mobility are mechanically unstable. A 1% vertical sodium dodecyl sulfate (SDS)-agarose gel electrophoresis (VAGE) system has been developed that allows titin (a protein with the largest known SDS subunit size of 3000-4000 kDa) to migrate over 10 cm in a approximately 13 cm resolving gel. Such migration gives clear and reproducible separation of titin isoforms. Proteins ranging in size from myosin heavy chain ( approximately 220 kDa) up to titin can be resolved on this gel system. Electroblotting of these very large proteins was nearly 100% efficient. This VAGE system has revealed two titin size variants in rabbit psoas muscle, two N2BA bands in rabbit cardiac muscle, and species differences between titins from rat and rabbit muscle. Agarose electrophoresis should be the method of choice for separation and blotting of proteins with very large subunit sizes. PMID- 12783445 TI - Anthracycline-dependent heat-induced transition from positive to negative supercoiled DNA. AB - The conformational stability of individual DNA topoisomers depends on the concentration of DNA intercalating drugs. To study the DNA-drug interaction, we used ethidium bromide (EtBr) and negative supercoiled pUC19 as a model system. The effects of two anthracyclines widely used in cancer therapy, daunorubicin (Dau) and doxorubicin (Doxo), and EtBr were compared. In spite of their different chemical structures and intercalation mode, all intercalating agents show similar effects on the conformational stability of supercoiled DNA. Our observations show that the studied intercalators have at least two main effects on the supercoiled DNA: (i) they decrease the level of negative supercoiling and, at certain concentrations, they may induce positive supercoiling in DNA; (ii) a temperature increase can cause a recovery of negative supercoiling in DNA. The conformational stability of plasmid DNA-drug complexes has been investigated by temperature gradient gel electrophoresis (TGGE). We demonstrate the suitability of TGGE for this purpose, because it offers a global view on DNA-drug complexes over a continuous range of temperature. Images of DNA plasmids adsorbed onto a substrate at different temperatures and drug concentrations were acquired by atomic force microscopy (AFM), allowing us to distinguish directly the conformation chirality assumed by the plasmid under different conditions confirming TGGE results. Our detection system allows to characterize unknown drugs and to determine their intercalating properties. PMID- 12783446 TI - Method for clone checking. AB - A new method for simple and fast clone checking is described. We combined the Pyrosequencing technology with a preprogrammed nucleotide dispensation strategy for fast analysis of DNA constructs. To test this method, the N-terminus region of plasmids constructed for the production of recombinant apyrase was analyzed. Of the ten plasmids tested, seven constructs were correct, two constructs showed one base deletion, and one construct showed deletion of a 195 bp fragment. The preprogrammed nucleotide dispensation strategy allowed the identification of the sequence downstream of the deletions. Thus, this method determines both the location and nature of possible artifacts. PMID- 12783447 TI - Cycling gradient capillary electrophoresis: a low-cost tool for high-throughput analysis of genetic variations. AB - In the present work, we introduce a new type of DNA variation detection. This method represents a transfer of melting gel technique onto multicapillary electrophoresis DNA sequencing instrument with further improvements to achieve maximum sample throughput while maintaining a high performance. The main improvement comes from application of cycling (revolving) temporal temperature gradient in place of a single-sweep gradient, commonly used in similar gel-based techniques. This improvement enables utilization of multiple-injection technique, in which multiple samples are injected into the same capillary (or sets of capillaries) separated by predefined time intervals of partial electrophoresis. The periodic oscillation of the temperature results in identical separation conditions of all samples injected in such series. Using this novel approach, we demonstrate a dramatic increase in separation throughput by turning a standard commercial 96-capillary array instrument into a semicontinuous flow mutation detection system capable to screen over 15 000 samples in 24 h of operation on a single 96-capillary commercial instrument. This represents a 10-fold increase in sample throughput over the current comparable technology. PMID- 12783448 TI - On-column polymer-imbedded graphite inlet electrode for capillary electrophoresis coupled on-line with flow injection analysis in a poly(dimethylsiloxane) interface. AB - A method for coupling an electrophoretic driven separation to a liquid flow, using conventional fused-silica capillaries and a soft polymeric interface is presented. A novel design of the electrode providing high voltage to the electrophoretic separation was also developed. The electrode consisted of a conductive polyimide/graphite imbedded coating immobilized onto the capillary electrophoresis (CE) column inlet. This integrated electrode gave the same separation performance as a commonly used platinum electrode. The on-column electrode also showed good electrochemical stability in chronoamperometric experiments. In addition, with this electrode design, the electrode position relative to the inlet end of the CE column will always be constant and well defined. The on-line flow injection analysis (FIA)-CE system was used with electrospray ionization (ESI)-time of flight (TOF)-mass spectrometry detection. The preparation of the PDMS (poly(dimethylsiloxane)) interface for FIA-CE is described in detail and used for initial tests of the on-column polymer-imbedded graphite inlet electrode. In this interface, a pressure-driven liquid flow, a make up CE electrolyte and a CE column inlet meet in a two-level cross (95 microm ID) in the PDMS structure, enabling independent flow characterization. PMID- 12783449 TI - A simple, rapid, and sensitive method for analysis of SYPRO Red labeled sodium dodecyl sulfate-protein complexes by capillary electrophoresis with laser-induced fluorescence. AB - We describe a segmental filling method for the analysis of SYPRO Red labeled sodium dodecyl sulfate (SDS)-proteins (SRSPs) by capillary electrophoresis-laser induced fluorescence (CE-LIF) with electroosmotic counterflow of poly(ethylene oxide) (PEO). It is shown that SDS and salt play a crucial role in determining the fluorescence intensity of the SRSP. Although the fluorimetric measurements reveal that the SRSPs fluoresce strongly in Tris-borate (TB) buffer containing 0.1% SDS and high concentrations of NaCl (100 mM), these conditions are not appropriate to CE in view of Joule heating. To overcome that impediment, we applied a plug of 0.1% SDS (1/5 to 1/3 of the injection volume) prior to injection of samples (0.64 microL) prepared in TB buffer containing 50 mM NaCl and SYPRO Red. When using a background electrolyte of 0.6% PEO in TB buffer containing NaCl, electroosmotic counterflow of the analytes allows one to concentrate large sample volumes (up to 1/3 of effective capillary length) in 21 min, with detection of 0.35 and 0.10 nM for bovine serum albumin and casein, respectively. With a linear dynamic range from 10 nM to 5 microM, this method provides the capability of determining the concentration of casein in cow's milk as 0.45 +/- 0.03 mM (n = 5). PMID- 12783450 TI - Electrophoretic separation and confocal laser-induced fluorescence detection at ultralow concentrations in constricted fused-silica capillaries. AB - Laser-induced fluorescence detection of labeled amino acids in a flowing stream at femtomolar (10(-15)M) concentrations was achieved by using a fused-silica capillary flow-cell comprising a constricted thin-walled detection region with inner diameters (IDs) ranging from 2 to 8 microm. The diameter of the constricted region was made to match a diffraction-limited focus of a uniphase transverse electromagnetic mode (TEM(00)) laser beam. Optimization of capillary dimensions and geometries (i.e., curvature, wall thickness, and outer-inner diameter ratio were performed in order to minimize cylindrical lensing of the focused laser beam. The fluorescence was collected in a confocal optical setup using a 1.3 numerical aperture (NA), 100x oil-immersion objective and a single-photon counting avalanche diode (SPAD). Under conditions of fluid flow, the constriction in the capillary forces all analytes to traverse across the laser probe volume, resulting in a high sampling efficiency. Fluorescein isothiocyanate-labeled glutamate (FITC-Glu) was electrophoretically separated and detected in capillaries having an ID of 2 microm at the constricted region with detection limits of 250 fM (signal-to-noise ratio (S/N) = 3) in the injected solution. PMID- 12783451 TI - Investigation of the mechanism of pH-mediated stacking of anions for the analysis of physiological samples by capillary electrophoresis. AB - Capillary electrophoresis has been widely used for the analysis of physiological samples such as plasma and microdialysate. However, sample destacking can occur during the analysis of these high-ionic strength samples, resulting in poor separation efficiency and reduced sensitivity. A technique termed pH-mediated stacking of anions (base stacking) has previously been developed to analyze microdialysate samples and achieve on-line preconcentration of analytes by following sample injection with an injection of sodium hydroxide. In this work, the mechanism of base stacking was investigated. Peak efficiency was shown to be a function of background electrolyte and sample ionic strength. Analytes representing several classes of compounds with a wide range of mobilities were used to study the effects of multiple parameters on sample stacking. The length of hydroxide injection required for stacking was shown to be dependent on analyte mobility and the type of amine background electrolyte used. Combinations of electrokinetic and hydrodynamic injections of sample and hydroxide were examined and it was concluded that although stacking could be achieved with several injection modes, electrokinetic injection of both sample and hydroxide was most effective for sample stacking. The mechanism of pH-mediated stacking for each of these modes is presented. PMID- 12783452 TI - Size-exclusion capillary electrochromatographic separation of polysaccharides using polymeric stationary phases. AB - We report the successful size-based separations of large, neutral polysaccharides using capillary electrochromatography (CEC). As the polysaccharides possessed little chromophore for photometric detection, two separate approaches were taken. In the first approach, indirect detection was combined with size-exclusion chromatography using a sulfonated polystyrene/divinylbenzene stationary phase. The separations were performed using a 300 A pore size stationary phase under aqueous conditions. Non-size based interactions were minimal using this material, resulting in an effective calibration range of molecular masses 180 to 112 000 g.mol(-1) for pullulans. In the second approach, the polysaccharides were derivatized with phenylisocyanate and were subsequently separated on columns made using a combination of high capacity ion-exchanger and a neutral polystyrene/divinylbenzene material of various pore sizes. The sulfonated ion exchange phase provided the electroosmotic flow, while the mixed pore size material provided the extended calibration range. The linear range for this primarily nonaqueous system using tetrahydrofuran was determined to be from molecular masses 738 to 404 000 g.mol(-1) of the original, untagged pullulan. This approach overcame the limited solubility issue associated with analysis of some polysaccharides. Analysis of pullulan and amylose samples by CEC correlated well with results obtained by conventional high-performance liquid chromatography (HPLC). The size-exclusion electrochromatographic separations provide an alternative mode for determining the relative molecular weights of polysaccharides with reduced sample and solvent consumption, as well as analysis times. PMID- 12783453 TI - Cyclodextrin-modified micellar electrokinetic chromatography for the analysis of Esterom, a topical product consisting of hydrolyzed benzoylecgonine in propylene glycol. AB - Esterom, a new drug currently in human clinical trials, is a mixture of compounds in a propylene glycol vehicle. It is being evaluated as a topical treatment to aid in the relief of muscle pain and to increase range of motion. Benzoylecgonine is the major component of Esterom and there are at least nine other minor constituents, including four hydroxypropyl esters that have multiple diasteriomers. The aim of the study was to develop a capillary electrophoresis method for the simultaneous separation of the main components in Esterom, including the multiple proposed diastereomers of the esters. Due to the complex sample composition, the use of micelles and cyclodextrins as buffer modifiers was evaluated. A cyclodextrin-modified micellar electrokinetic chromatography method was able to determine 7 of the 8 UV-active Esterom components, with baseline separation of 7 of the 10 diastereomers of the hydroxypropyl esters. PMID- 12783454 TI - Chiral analysis of amphetamine-type stimulants using reversed-polarity capillary electrophoresis/positive ion electrospray ionization tandem mass spectrometry. AB - Reversed-polarity (RP) capillary electrophoresis/positive ion electrospray ionization mass spectrometry (CE-ESI+ MS) and tandem mass spectrometry (MS/MS) were utilized for simultaneous chiral separation of nine amphetamine-type stimulants (ATS) (dl-norephedrine, dl-norpseudoephedrine, dl-ephedrine, dl pseudoephedrine, dl-amphetamine, dl-methamphetamine, dl methylenedioxyamphetamine, dl-methylenedioxymethamphetamine, and dl methylenedioxyethylamphetamine). Using highly sulfated gamma-cyclodextrin (SU(XIII)-gamma-CD) as a chiral selector, the nine ATS were completely separated within 50 min. The migrated ATS-CD complex was dissociated at the ESI interface, and only ATS molecules went into the MS detector so that all 18 individual enantiomers were identified by their mass spectra. The detection limit of MS/MS was 10 times more sensitive than those for single MS. Seized d-methamphetamine hydrochloride samples dissolved at high concentration (20 mg/mL) were analyzed. Impurities originating in the precursor such as l-ephedrine and d-pseudoephedrine were detected and identified by tandem mass spectra. PMID- 12783455 TI - Improved resolution with microchip-based enhanced field inversion electrophoresis. AB - We present an improvement of the field inversion electrophoresis (FIE) method in which the passage of sample such as DNA back and forth within a short length of a microchannel can provide a similar resolution to that of a significantly longer microchannel. In constant field FIE the application of an alternating potential (e.g., +/- V) over short periods of time (e.g., several Hz) can provide enhanced separations of DNA fragments. In contrast, the present method consists of a series of separations, each of much longer duration, under high and low fields in such a way that the resolution is enhanced. This method is readily modeled and allows improved resolution to be obtained from extremely short microchannels (e.g., 8 mm) while requiring relatively low applied voltages (e.g., less than 600 V). An additional advantage is that this method can allow for the same equipment to be used in a rapid, low-resolution mode or in a slower, high-resolution mode through what might be referred to as an automated "zoom" capability. We believe that this method may facilitate the integration of microfluidic devices and microelectronic devices by allowing these devices to be of a similar small scale (< 1 cm). PMID- 12783456 TI - A versatile electrophoresis system for the analysis of high- and low-molecular weight proteins. AB - A new, versatile, multiphasic buffer system for high-resolution sodium dodecyl sulfate-polyacrylamide gel electrophoresis of proteins in the relative molecular weight range of 300 000-3000 Da is described. The system, based on the theory of multiphasic zone electrophoresis, allows complete stacking and destacking of proteins in the above M(r) range. The buffer system uses taurine and chloride as trailing and leading ion, respectively, and Tris, at a pH close to its pK(a), as the buffering counterion. Coupled with limited variation in the acrylamide concentration, this electrophoresis system allows to tailor the resolution in the 6-200 kDa M(r) range, with minimal difficulties in the post electrophoretic identification processes. PMID- 12783458 TI - Profiling stage-dependent changes of protein expression in Caenorhabditis elegans by mass spectrometric proteome analysis leads to the identification of stage specific marker proteins. AB - Proteome maps obtained by synchronization of the wild-type Caenorhabditis elegans development reflected stage-dependent molecular differences and revealed dynamic cytoskeletal processes during ontogenesis. Distinct protein spots that may function as molecular markers for the corresponding developmental stages were mass spectrometrically identified. The amount of the Cu(2+)- Zn(2+) superoxide dismutase (CE23550) and an aspartyl proteinase (CE21681) was highest in the first larval stage (L1) and decreased during the ontogenesis from the first larval stage to the adult. Tropomyosin III (CE29059) was prominently present in the first and second larval stage (L1/L2). Abundances of actin 1 or 4 (CE12358 or CE13148) and tropomyosin I (CE28782) were particularly high in multiple spots in the third larval stage (L3). Interestingly, the amount of DIM-1 protein (CE27706), reflected by two spots, was the lowest in this stage. A particular splicing factor (CE31089) was detected only in the fourth larval stage (L4), whereas a spot with high abundance representing the cuticle collagen (CE02272) was only found highly expressed in adult animals (A). In addition, a Ca(2+) binding protein (CE12368) and one protein spot which has not yet been identified, both reached their maximal spot intensities in the adult stage (A). Moreover, the ASP-1, CCT-5, GPD-1, GPD-2, HSP-6, HSP-16.2, IFB-2, LEC-2, LIN-53, LMN-1, MDH-1, NUD-1, RPA-0, RSP-12, SOD-1, TBB-1, TBB-2, TMY-1, UNC-60, and VIT-2 proteins for which mutants are available and two still unidentified protein spots which were present in all developmental stages, have been reproducibly localized in proteome maps of distinct ontogenesis states. PMID- 12783457 TI - An optimized method for the isolation and identification of membrane proteins. AB - The purpose of this study was to develop a protocol suitable for membrane protein extraction from limited starting material and to identify appropriate conditions for two-dimensional (2-D) gel electrophoresis. We used A549 cells, a human alveolar type II cell line, and evaluated three protein extraction methods based on different separation principles, namely protein solubility, detergent-based and density-based organelle separation. Detergent-based extraction achieved the highest yield with 14.64% +/- 2.35 membrane proteins but sequential extraction with 7.35% +/- 0.78 yield and centrifugal extraction with 4.1% +/- 0.54 yield produced the purest fractionation of membrane proteins. Only the sequential and the detergent-based extraction proved suitable for small volumes of starting material. We identified annexin I + II, electron transfer flavoprotein beta chain, H(+)-transporting ATP synthase, mitofilin and protein disulfide isomerase A3 as membrane and cytokeratin 8 + 18, actin and others as soluble proteins using matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) analysis and started to map the A549 cell proteome. Our data suggest that membrane proteins can be extracted efficiently from small samples using a simple sequential protein extraction method. They can be separated and identified successfully using optimized conditions in 2-D gel electrophoresis. The presented methods will be useful for further investigations of membrane proteins of alveolar and bronchial carcinomas. PMID- 12783459 TI - Spot volume vs. amount of protein loaded onto a gel: a detailed, statistical comparison of two gel electrophoresis systems. AB - The long-term goal of this research program is to clarify the molecular mechanisms that participate in the formation of human pituitary macroadenomas. One approach to that goal is to characterize the differentially expressed proteins that are found by a comparison of the proteomes of control pituitary vs. macroadenoma tissues. In order to accurately perform a comparative proteomics study, based on the combination of two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and PDQuest 2-D analysis software, a reproducible 2-DE separation system with a wide linear dynamic measure range is needed. A typical horizontal system is the Multiphor II system that analyzes one gel at a time, using a precast gradient gel (180 x 245 x 0.5 mm); a typical vertical system is the Dodeca system that analyzes up to 12 gels at a time on a single-concentration gel (190 x 205 x 1.0 mm). We have evaluated (Zhan and Desiderio, Electrophoresis 2003, 24, 1834-1846) the spatial and quantitative reproducibility of the two second-dimensional gel systems to separate a human pituitary proteome; that study showed a higher reproducibility for the Dodeca gel system. This present study investigated the relationship between the spot volume and the amount of protein loaded onto the gel for those two 2-D systems. The results demonstrated that the Dodeca gel system provides a wider linear dynamic range to measure the changes in the protein abundance in pituitary proteome. PMID- 12783460 TI - Differences in the spatial and quantitative reproducibility between two second dimensional gel electrophoresis systems. AB - Two-dimensional polyacrylamide gel electrophoresis (PAGE), together with 2-D gel electrophoresis (GE) analysis software, is a common technique to analyze a complex proteome. In order to accurately locate the differentially expressed proteins in human pituitary macroadenoma tissues in our long-term research program to clarify the molecular mechanisms of macroadenoma formation, a reproducible separation system is needed. An immobilized pH-gradient dry gel strip (IPG strip) has been extensively used for first-dimensional isoelectric focusing (IEF), and has achieved a high degree of reproducibility in the IEF direction. For the second dimension (SDS-PAGE), different types of gel systems are available, including horizontal vs. vertical gel systems, and gradient vs. constant-percentage gels. A typical horizontal system is the Multiphor II system that analyzes one gel at a time, using a precast gradient gel (180 x 245 x 0.5 mm), and a typical vertical system is the Dodeca system, which analyzes up to 12 gels at a time, using usually a single-concentration gel (190 x 205 x 1 mm). The present study evaluated the spatial and quantitative reproducibility of the two systems for the separation of the complex human pituitary proteome. PDQuest software was used to analyze the digitized gel-image data, and SPSS statistical software was used to analyze the data. The results demonstrated a high percentage (>99%) of protein-spot matches within each electrophoretic system. The Dodeca gel system demonstrated better between-gel reproducibility for spot position, higher resolution in the Sodium dodecyl sulfate (SDS)-PAGE direction, lower gel background, better spot quality, and higher reproducibility of the spot volume. PMID- 12783461 TI - A rat brain protein expression map including cytosolic and enriched mitochondrial and microsomal fractions. AB - Proteomics is a powerful tool to screen brain protein expression but the methodology is hampered by low abundance of proteins or compartmentalization or overload of high-abundance proteins. It was therefore the aim of the study to determine the expression of brain proteins by using enriched cellular subfractions and pre-electrophoretic chromatographical separation of brain homogenates. We used two-dimensional electrophoresis with subsequent matrix assisted laser desorption/ionization (MALDI) detection and characterization of brain proteins. Subfractionation into cytosolic, mitochondrial and microsomal compartments was performed by ultracentrifugation. Pre-electrophoretic fractionation of the cytosolic fractions was carried out by ion exchange column chromatography. We detected and identified a large series of 437 proteins in rat brain and have shown proteins specific for the individual subcellular compartments. These proteins included housekeeping, signaling, cytoskeletal, intermediary metabolism, antioxidant proteins on the one and neuron and synaptosomal specific proteins on the other hand. Using fractionations of brain homogenates we were able to improve the power of the method on forming the basis for brain protein expressional studies and providing a reference map as a powerful tool for the neuroscientist. PMID- 12783462 TI - Proteomic profiling of pancreatic ductal carcinoma cell lines treated with trichostatin-A. AB - A pancreatic adenocarcinoma cell line (Paca44) was treated with trichostatin-A (TSA), a potent inhibitor of histone deacetylases, in order to evaluate the effect of this drug on protein expression. Master maps of control and treated Paca44 cells were generated by analysis with the PDQuest software. The comparison between such maps showed up- and downregulation of 51 polypeptide chains, out of a total of 700 spots detected by a medium-sensitivity stain, micellar Coomassie Brilliant Blue. Fingerprinting by matrix-assisted laser desorption/ionization time of flight (MALDI-TOF)-mass spectrometry analysis enabled the identification of 22 of these spots. Among these proteins, of particular interest are the two downregulated proteins nucleophosmin and translationally controlled tumor protein, as well as the upregulated proteins programmed cell death protein 5 (also designated as TFAR19) and stathmin (oncoprotein 18). The modulation of these four proteins is consistent with our observation that TSA is able to inhibit cell growth of Paca44 by causing cell cycle arrest at the G2 phase and apoptotic cell death. PMID- 12783481 TI - Expression of a synthetic copy of the bovine chymosin gene in Aspergillus awamori from constitutive and pH-regulated promoters and secretion using two different pre-pro sequences. AB - A copy of the bovine chymosin gene (chy) with a codon usage optimized for its expression in Aspergillus awamori was constructed starting from synthetic oligonucleotides. To study the ability of this filamentous fungus to secrete bovine prochymosin, two plasmids were constructed in which the transcriptional, translational, and secretory control regions of the A. nidulans gpdA gene and pepB genes were coupled to either preprochymosin or prochymosin genes. Secretion of a protein enzymatically and immunologically indistinguishable from bovine chymosin was achieved in A. awamori transformants with each of these constructions. In all cases, the primary translation product (40.5 kDa) was self processed to a mature chymosin polypeptide having a molecular weight of 35.6 kDa. Immunological assays indicated that most of the chymosin was secreted to the extracellular medium. Hybridization analysis of genomic DNA from chymosin transformants showed chromosomal integration of prochymosin sequences and, in some transformants, multiple copies of the expression cassettes were observed. Expression from the gpdA promoter was constitutive, whereas expression from the pepB promoter was strongly influenced by pH. A very high expression from the pepB promoter was observed during the growth phase. The A. awamori pepB gene terminator was more favorable for chymosin production than the S. cerevisiae CYC1 terminator. PMID- 12783482 TI - Experimental investigation of the external nitrification biological nutrient removal activated sludge (ENBNRAS) system. AB - A systematic lab-scale experimental investigation is reported for the external nitrification (EN) biological nutrient removal (BNR) activated sludge (ENBNRAS) system, which is a combined fixed and suspended medium system. The ENBNRAS system was proposed to intensify the treatment capacity of BNR-activated sludge (BNRAS) systems by addressing two difficulties often encountered in practice: (a) the long sludge age for nitrification requirement; and (b) sludge bulking. In the ENBNRAS system, nitrification is transferred from the aerobic reactor in the suspended medium activated sludge system to a fixed medium nitrification system. Thus, the sludge age of the suspended medium activated sludge system can be reduced from 20 to 25 days to 8 to 10 days, resulting in a decrease in reactor volume per ML wastewater treated of about 30%. Furthermore, the aerobic mass fraction can also be reduced from 50% to 60% to <30% and concommitantly the anoxic mass fraction can be increased from 25% to 35% to >55% (if the anaerobic mass fraction is 15%), and thus complete denitrification in the anoxic reactors becomes possible. Research indicates that both the short sludge age and complete denitrification could ameliorate anoxic aerobic (AA) or low food/microorganism (F/M) ratio filamentous bulking, and hence reduce the surface area of secondary settling tanks or increase the treatment capacity of existing systems. The lab scale experimental investigations indicate that the ENBNRAS system can obtain: (i) very good chemical oxygen demand (COD) removal, even with an aerobic mass fraction as low as 20%; (ii) high nitrogen removal, even for a wastewater with a high total kjeldahl nitrogen (TKN)/COD ratio, up to 0.14; (iii) adequate settling sludge (diluted sludge volume index [DSVI] <100 mL/g); and (iv) a significant reduction in oxygen demand. PMID- 12783483 TI - Lipase-catalyzed ethanolysis of fish oils: multi-response kinetics. AB - The kinetics of the lipase-catalyzed (Pseudomonas cepacia) ethanolysis of fish oil has been studied in a batch reactor using menhaden oil, tuna oil, and acylglycerol mixtures derived from menhaden oil. Multi-response models derived from a generalized Michaelis-Menten mechanism were developed to describe the rates of formation of ethyl esters of the primary fatty acids present in the precursor oil. A first-order model for deactivation of the lipase was fit simultaneously to one of the data sets. PMID- 12783484 TI - Properties of a novel magnetized alginate for magnetic resonance imaging. AB - Implanting recombinant cells encapsulated in alginate microcapsules to secrete therapeutic proteins has been proven clinically effective in treating several murine models of human diseases. However, once implanted, these microcapsules cannot be assessed without invasive surgery. We now report the preparation and characterization of a novel ferrofluid to render these microcapsules visible with magnetic resonance imaging (MRI). The ferrofluid was prepared as a colloidal iron oxide stabilized in water by alginate. The presence of iron particles in the ferrofluid was verified with chemical titration, dynamic light scattering, and magnetization measurement. The microcapsules fabricated with various concentrations of the ferrofluid in the core, or on the surface of alginate microcapsules, or both, all produced microcapsules with smooth surfaces as shown with light and scanning electron microscopy. However, at the nanoscale level, as revealed with atomic force microscopy, the ferrofluid-fabricated microcapsules demonstrated increased granularity, particularly when the ferrofluid was used to laminate the surface. From the force spectroscopy measurements, these modified microcapsules showed increasing surface rigidity in the following order: traditional alginate < ferrofluid in the core < ferrofluid on the surface. Although the mechanical stability of low-concentration ferrofluid (0.1%) microcapsules was reduced, increasing concentrations, up to 20%, were able to improve stability. When these ferrofluid microcapsules were examined with MRI, their T(2) relaxation time was reduced, thereby producing increased contrast readily detectable with MRI, whereas the traditional alginate microcapsules showed no difference when compared with water. In conclusion, such ferrofluid enhanced alginate is suitable for fabricating microcapsules that offer the potential for in vivo tracking of implanted microcapsules without invasive surgery. PMID- 12783485 TI - Model-based analysis of anaerobic acetate uptake by a mixed culture of polyphosphate-accumulating and glycogen-accumulating organisms. AB - An increasing number of studies shows that the glycogen-accumulating organisms (GAOs) can survive and may indeed proliferate under the alternating anaerobic/aerobic conditions found in EBPR systems, thus forming a strong competitor of the polyphosphate-accumulating organisms (PAOs). Understanding their behaviors in a mixed PAO and GAO culture under various operational conditions is essential for developing operating strategies that disadvantage the growth of this group of unwanted organisms. A model-based data analysis method is developed in this paper for the study of the anaerobic PAO and GAO activities in a mixed PAO and GAO culture. The method primarily makes use of the hydrogen ion production rate and the carbon dioxide transfer rate resulting from the acetate uptake processes by PAOs and GAOs, measured with a recently developed titration and off-gas analysis (TOGA) sensor. The method is demonstrated using the data from a laboratory-scale sequencing batch reactor (SBR) operated under alternating anaerobic and aerobic conditions. The data analysis using the proposed method strongly indicates a coexistence of PAOs and GAOs in the system, which was independently confirmed by fluorescent in situ hybridization (FISH) measurement. The model-based analysis also allowed the identification of the respective acetate uptake rates by PAOs and GAOs, along with a number of kinetic and stoichiometric parameters involved in the PAO and GAO models. The excellent fit between the model predictions and the experimental data not involved in parameter identification shows that the parameter values found are reliable and accurate. It also demonstrates that the current anaerobic PAO and GAO models are able to accurately characterize the PAO/GAO mixed culture obtained in this study. This is of major importance as no pure culture of either PAOs or GAOs has been reported to date, and hence the current PAO and GAO models were developed for the interpretation of experimental results of mixed cultures. The proposed method is readily applicable for detailed investigations of the competition between PAOs and GAOs in enriched cultures. However, the fermentation of organic substrates carried out by ordinary heterotrophs needs to be accounted for when the method is applied to the study of PAO and GAO competition in full-scale sludges. PMID- 12783486 TI - Kinetic modeling of the light-dependent photosynthetic activity of the green microalga Chlorella vulgaris. AB - Light-dependent photosynthesis of Chlorella vulgaris was investigated by using a novel photosynthesis measurement system that could cover wide ranges of incident light and cell density and reproduce accurate readings. Various photosynthesis models, which have been reported elsewhere, were classified and/or reformulated based upon the underlying hypotheses of the light dependence of the algal photosynthesis. Four types of models were derived, which contained distinct light related variables such as the average or local photon flux density (APFD or LPFD) and the average or local photon absorption rate (APAR or LPAR). According to our experimental results, the LPFD and LPAR models could predict the experimental data more accurately although the APFD and APAR models have been widely used for the kinetic study of microalgal photosynthesis. PMID- 12783487 TI - Inhibitory effect of carbon dioxide on the fed-batch culture of Ralstonia eutropha: evaluation by CO2 pulse injection and autogenous CO2 methods. AB - In order to see the effect of CO(2) inhibition resulting from the use of pure oxygen, we carried out a comparative fed-batch culture study of polyhydroxybutyric acid (PHB) production by Ralstonia eutropha using air and pure oxygen in 5-L, 30-L, and 300-L fermentors. The final PHB concentrations obtained with pure O(2) were 138.7 g/L in the 5-L fermentor and 131.3 g/L in the 30-L fermentor, which increased 2.9 and 6.2 times, respectively, as compared to those obtained with air. In the 300-L fermentor, the fed-batch culture with air yielded only 8.4 g/L PHB. However, the maximal CO(2) concentrations in the 5-L fermentor increased significantly from 4.1% (air) to 15.0% (pure O(2)), while it was only 1.6% in the 30-L fermentor with air, but reached 14.2% in the case of pure O(2). We used two different experimental methods for evaluating CO(2) inhibition: CO(2) pulse injection and autogenous CO(2) methods. A 10 or 22% (v/v) CO(2) pulse with a duration of 3 or 6 h was introduced in a pure-oxygen culture of R. eutropha to investigate how CO(2) affects the synthesis of biomass and PHB. CO(2) inhibited the cell growth and PHB synthesis significantly. The inhibitory effect became stronger with the increase of the CO(2) concentration and pulse duration. The new proposed autogenous CO(2) method makes it possible to place microbial cells under different CO(2) level environments by varying the gas flow rate. Introduction of O(2) gas at a low flow rate of 0.42 vvm resulted in an increase of CO(2) concentration to 30.2% in the exit gas. The final PHB of 97.2 g/L was obtained, which corresponded to 70% of the PHB production at 1.0 vvm O(2) flow rate. This new method measures the inhibitory effect of CO(2) produced autogenously by cells through the entire fermentation process and can avoid the overestimation of CO(2) inhibition without introducing artificial CO(2) into the fermentor. PMID- 12783488 TI - Effects of buffering conditions and culture pH on production rates and glycosylation of clinical phase I anti-melanoma mouse IgG3 monoclonal antibody R24. AB - R24, a mouse IgG3 monoclonal antibody (MAb) against ganglioside GD3 (Neu5Acalpha8Neu5Acalpha3Gal beta4Glcbeta1Cer), can block tumor growth as reported in a series of clinical trials in patients with metastatic melanoma. The IgG molecule basically contains an asparagine-linked biantennary complex type oligosaccharide on the C(H)2 domain of each heavy chain, which is necessary for its in vivo effector function. The purpose of this study was to investigate the biotechnological production and particularly the glycosylation of this clinically important MAb in CO(2)/HCO(3) (-) (pH 7.4, 7.2, and 6.9) and HEPES buffered serum free medium. Growth, metabolism, and IgG production of hybridoma cells (ATCC HB 8445) were analyzed on a 2-L bioreactor scale using fed-batch mode. Specific growth rates (mu) and MAb production rates (q(IgG)) varied significantly with maximum product yields at pH 6.9 (q(IgG) = 42.9 microg 10(-6) cells d(-1), mu = 0.30 d(-1)) and lowest yields in pH 7.4 adjusted batches (q(IgG) = 10.8 microg 10(-6) cells d(-1), mu = 0.40 d(-1)). N-glycans were structurally characterized by high pH anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD), matrix-assisted laser desorption ionization time-of-flight (MALDI TOF), and electrospray-ionization quadrupole time-of-flight (ESI-QTOF) mass spectrometry (MS). The highest relative amounts of agalacto and monogalacto biantennary complex type oligosaccharides were detected in the pH 7.2 (46% and 38%, respectively) and pH 6.9 (44% and 40%, respectively) cultivations and the uppermost quantities of digalacto (fully galactosylated) structures in the pH 7.4 (32%) and the HEPES (26%) buffered fermentation. In the experiments with HEPES buffering, antibodies with a molar Neu5Ac/Neu5Gc ratio of 3.067 were obtained. The fermentations at pH 7.2 and 6.9 resulted in almost equal molar Neu5Ac/Neu5Gc ratios of 1.008 and 0.985, respectively, while the alkaline shift caused a moderate overexpression of Neu5Ac deduced from the Neu5Ac/Neu5Gc quotient of 1.411. Different culture buffering gave rise to altered glycosylation pattern of the MAb R24. Consequently, a detailed molecular characterization of MAb glycosylation is generally recommended as a part of the development of MAbs for targeted in vivo immunotherapy to assure biochemical consistency of product lots and oligosaccharide-dependent biological activity. PMID- 12783489 TI - Multicriteria optimization of biochemical systems by linear programming: application to production of ethanol by Saccharomyces cerevisiae. AB - In this study we present a method for simultaneous optimization of several metabolic responses of biochemical pathways. The method, based on the use of the power law formalism to obtain a linear system in logarithmic coordinates, is applied to ethanol production by Saccharomyces cerevisiae. Starting from an experimentally based kinetic model, we translated it to its power law equivalent. With this new model representation, we then applied the multiobjective optimization method. Our intent was to maximize ethanol production and minimize each of the internal metabolite concentrations. To ensure cell viability, all optimizations were carried out under imposed constraints. The different solutions obtained, which correspond to alternative patterns of enzyme overexpression, were implemented in the original model. We discovered few discrepancies between the S system-optimized steady state and the corresponding optimized state in the original kinetic model, thus demonstrating the suitability of the S-system representation as the basis for the optimization procedure. In all optimized solutions, the ATP level reached its maximum and any increase in its activity positively affected the optimization process. This work illustrates that in any optimization study no single criteria is of general application being the multiobjective and constrained task the proper way to address it. It is concluded that the proposed multiobjective method can serve to carry out, in a single study, the general pattern of behavior of a given metabolic system with regard to its control and optimization. PMID- 12783490 TI - Production of savinase and population viability of Bacillus clausii during high cell-density fed-batch cultivations. AB - The growth and product formation of a Savinase-producing Bacillus clausii were investigated in high-cell-density fed-batch cultivations with both linear and exponential feed profiles. The highest specific productivity of Savinase was observed shortly after the end of the initial batch phase for all feed profiles applied and, in addition, there was a time-dependent decrease in specific productivity. The specific glucose uptake rate increased with time for constant specific growth rate indicating that the maintenance requirements increased with time, possibly due to a decreasing K(+) concentration. The physiological state of the cells was monitored during the cultivations using a flow cytometry assay based on the permeability of the cell membrane to propidium iodide. In the latter parts of the fed-batch cultures with a linear feed profile, a large portion of the cell population was found to have a permeable membrane, indicating a large percentage of dead cells. By assuming that only cells with a nonpermeable membrane contributed to growth and product formation, the physiological properties of this subpopulation were calculated. PMID- 12783491 TI - Continuous cultivations of a Penicillium chrysogenum strain expressing the expandase gene from Streptomyces clavuligerus: Kinetics of adipoyl-7 aminodeacetoxycephalosporanic acid and byproduct formations. AB - The production kinetics of a transformed strain of Penicillium chrysogenum expressing the expandase gene from Streptomyces clavuligerus was investigated in chemostat cultivations. The recombinant strain produces adipoyl-7 aminodeacetoxycephalosporanic acid (ad-7-ADCA) as the major product; however, during the cultivations, the appearance of a major unknown and poorly secreted product was observed. Investigations using high-performance liquid chromatography (HPLC) and liquid chromatography-mass spectroscopy (LC-MS) showed that this byproduct has a six-membered dihydrothiazine ring, which is characteristic for cephalosporins. The byproduct may be formed via isopenicillin N by as-yet unknown mechanisms, but involving expandase. It is likely that the unknown compound (UC) is deacetoxycephalosporin C (DAOC). Investigation of the instability of the various beta-lactams produced showed higher instability for compounds with a five membered thiazolidine ring than those with a six-membered dihydrothiazine ring. Furthermore, secretion of products and byproducts was shown to be quite different. The productivity was studied as a function of the dilution rate in the range 0.015 to 0.090 h(-1). The specific productivity of total beta-lactams was compared with that of the penicillin-G-producing host strain, and it was found to be lower at dilution rates of <0.06 h(-1). Quantification of the fluxes through the pathway leading to ad-7-ADCA showed a decrease in flux toward ad-7-ADCA, and an increase in flux toward UC as the dilution rate increased. Northern analysis of the biosynthetic genes showed that expression of the enzymes involved in the ad-7-ADCA pathway decreased as the dilution rate increased. PMID- 12783495 TI - Conformation-selective laser chemistry. PMID- 12783492 TI - Continuous cultivations of a Penicillium chrysogenum strain expressing the expandase gene from Streptomyces clavuligerus: Growth yields and morphological characterization. AB - The growth stoichiometry of a Penicillium chrysogenum strain expressing the expandase gene from Streptomyces clavuligerus was determined in glucose-limited chemostat cultivations using a chemically defined medium. This strain produces adipoyl-7-aminodeacetoxycephalosporanic acid (ad-7-ADCA) when it is fed with adipic acid. The biomass yield and maintenance coefficients for the strain were similar to those found for penicillin-producing strains of Penicillium chrysogenum. The maximum specific growth rate in the chemostat was found to be 0.11 h(-1). Metabolic degradation of adipate was found to take place in significant amounts only at dilution rates below 0.03 h(-1). After three to five residence times, adipate degradation and ad-7-ADCA production disappeared, and this allowed determination of the biomass yield coefficient on adipate. The morphology was measured at different dilution rates and the mean total hyphal length and mean number of tips both increased with an increase in dilution rate from 0.015 to 0.065 h(-1). Both variables decreased when the dilution rate was increased above 0.065 h(-1). A correlation between mean total hyphal length and productivity of ad-7-ADCA was found. PMID- 12783496 TI - Ketenes in polymer-assisted synthesis. AB - Since its inception, ketene chemistry has developed into a unique and well established source of useful transformations for conventional synthetic organic chemistry. It is, therefore, not surprising that soon after their movement from the realm of peptide and peptoid libraries to that of small molecules, combinatorial chemists have sought the benefits of ketene chemistry to satisfy their own synthetic needs. The ability of these versatile molecules to undergo reactions with nucleophiles, and to participate in cycloadditions and cyclocondensations, has been utilized for the preparation of diverse heterocyclic compounds, and has added to the advantages of polymer-assisted synthesis for rapid purification. Different types of ketenes and different methods for their generation have been involved, which illustrates the potential diversity of the chemistry. There is now a better grasp of the effect of the fragility of these sometimes transient molecules on the reactions involving solid supports, and this augurs well for the application of some of the more recent developments in ketene chemistry to the generation of small-molecule libraries. PMID- 12783497 TI - Higher-order organization by mesoscale self-assembly and transformation of hybrid nanostructures. AB - The organization of nanostructures across extended length scales is a key challenge in the design of integrated materials with advanced functions. Current approaches tend to be based on physical methods, such as patterning, rather than the spontaneous chemical assembly and transformation of building blocks across multiple length scales. It should be possible to develop a chemistry of organized matter based on emergent processes in which time- and scale-dependent coupling of interactive components generate higher-order architectures with embedded structure. Herein we highlight how the interplay between aggregation and crystallization can give rise to mesoscale self-assembly and cooperative transformation and reorganization of hybrid inorganic-organic building blocks to produce single-crystal mosaics, nanoparticle arrays, and emergent nanostructures with complex form and hierarchy. We propose that similar mesoscale processes are also relevant to models of matrix-mediated nucleation in biomineralization. PMID- 12783498 TI - A virosome-mimotope approach to synthetic vaccine design and optimization: synthesis, conformation, and immune recognition of a potential malaria-vaccine candidate. PMID- 12783499 TI - Magnetically amplified DNA assays (MADA): sensing of viral DNA and single-base mismatches by using nucleic acid modified magnetic particles. PMID- 12783500 TI - Expanding the functional group compatibility of small-molecule microarrays: discovery of novel calmodulin ligands. PMID- 12783501 TI - HR22C16: a potent small-molecule probe for the dynamics of cell division. PMID- 12783502 TI - Crystallization of an ordered mesoporous Nb-Ta oxide. PMID- 12783503 TI - Zwitterionic relatives to the classic [(P-P)Rh(solv)2]+ ions: neutral catalysts active for H-E bond additions to olefins (E=C, Si, B). PMID- 12783504 TI - Lewis acid catalyzed room-temperature Michaelis-Arbuzov rearrangement. PMID- 12783505 TI - Insertion of thiocyanogen into a P-C bond and subsequent formation of polysulfide derivatives. PMID- 12783506 TI - The first X-ray crystal structures of halogenated [70]fullerene: C70Br10 and C70Br10.3 Br2. PMID- 12783507 TI - Ringing the changes: a remarkable carbene-free synthesis of difluorocyclopropenes. PMID- 12783508 TI - Parallel sheet secondary structure in beta-peptides. PMID- 12783509 TI - IPy2BF4-promoted intramolecular addition of masked and unmasked anilines to alkynes: direct assembly of 3-iodoindole cores. PMID- 12783510 TI - Aqueous catalytic Pauson-Khand-type reactions of enynes with formaldehyde: transfer carbonylation involving an aqueous decarbonylation and a micellar carbonylation. PMID- 12783511 TI - Entropically driven ring-opening-metathesis polymerization of macrocyclic olefins with 21-84 ring atoms. PMID- 12783512 TI - A soft-chemical method to synthesize Lewis acid surfaces of aluminum oxide. PMID- 12783513 TI - Determination of the configuration of an archaea membrane lipid containing cyclopentane rings by total synthesis. PMID- 12783514 TI - [Ga9(CMe3)9]*--a persistent cluster radical anion, boron-analogous chemistry with the heavier homologue gallium. PMID- 12783515 TI - Site-directed alteration of RNA sequence mediated by an engineered twin ribozyme. PMID- 12783517 TI - Kinetics of weak distance-dependent hole transfer in DNA by adenine-hopping mechanism. AB - The kinetics of hole transfer in DNA by adenine-hopping mechanism was investigated by the combined pulse radiolysis-laser flash photolysis method. The hole transfer from Ptz*+* to oxG across the (A)n-bridge preceded by the A-hopping mechanism and the weak distance-dependent hole transfer with the rates faster than 108 s-1 over the distance range of 7-22 A was demonstrated. In contrast, hole transfer from oxG*+ to Ptz followed the single-step super exchange mechanism. Thus, two different processes for the hole transfer across the identical (A)n-bridge in DNA have been demonstrated. The results clearly show that the mechanism of hole transfer in DNA strongly depends on the redox nature of the oxidant, whether it produces only G*+ or both A*+ and G*+. PMID- 12783518 TI - Osmium replica of mesoporous silicate MCM-48: efficient and reusable catalyst for oxidative cleavage and dihydroxylation reactions. AB - A three-dimensional networked osmium nanomaterial (N-Os) was prepared by a thermal decomposition of Os3(CO)12 within mesopores of MCM-48. The novel N-Os species shows high catalytic activity and excellent reusability in the oxidation reactions of unsaturated compounds under mild conditions. PMID- 12783519 TI - A concise approach to structurally diverse beta-amino acids. AB - We have demonstrated that the high yields and selectivities of 1,3-dipolar cycloadditions can be translated into facile stereoselective syntheses of a diverse array of beta-amino acids, key components of bioactive natural products, beta-lactams, and peptidomimetics. Simply by selecting different combinations of three readily available starting materials (an oxime, a chiral allylic alcohol, and a nucleophile), we used the reaction sequence to prepare four different beta amino acid structural types with a variety of substitution patterns in good overall yield. Of particular note is the use of this approach to prepare highly substituted beta-amino acids not readily accessible by previously reported methodologies. This will pave the way for future studies of the structure and function of this important class of molecules. PMID- 12783520 TI - Fabrication of assembled virus nanostructures on templates of chemoselective linkers formed by scanning probe nanolithography. AB - We have developed a multistep route to the fabrication of virus assembled nanostructures with chemoselective protein-to-surface linkers synthesized by an efficient solid-phase method. These linkers were used to create patterns of 30-to 50-nm-width-lines by scanning probe nanolithography. Genetically modified cow pea mosaic virus with unique cysteine residues at specific locations on their capsomers were assembled through covalent linkage on these patterns. The morphology of the assembled structures on these line patterns characterized by atomic force microscopy was found to be strongly influenced by the intervirion interactions. PMID- 12783521 TI - Different binding orientations for the same agonist at homologous receptors: a lock and key or a simple wedge? AB - Using unnatural amino acid mutagenesis, the binding site for serotonin at the novel Caenorhabditis elegans receptor MOD-1 has been probed. As with the closely related serotonin receptor 5-HT3, MOD-1 makes use of a strong cation-pi interaction between the ammonium of serotonin and the indole side chain of a tryptophan. However, the specific Trp used by MOD-1 is different from that used for 5-HT3 (and the nAChR), aligning with a residue more than 40 amino acids distant in sequence space and on a different "loop" of the agonist binding site. This suggests a significant rearrangement of the ligand on binding these two closely related receptors. It is suggested that, unlike enzymes, receptors and other signaling molecules may need only to deliver an agonist to a general binding region, rather than establishing precise drug-receptor interactions. PMID- 12783522 TI - Novel selective inhibitors of the interaction of individual nuclear hormone receptors with a mutually shared steroid receptor coactivator 2. AB - Nuclear hormone receptor (NR) signaling, currently a therapeutic target in multiple diseases, involves an ordered series of protein interactions to regulate transcription in response to changing hormone levels. Later steps in the process of ligand-dependent signaling are driven by a highly conserved interaction between the NRs and the steroid receptor coactivators (SRCs) that is effected by a conserved interaction motif (L1XXL2L3), known as an NR box. Using computational design and combinatorial chemistry, we have produced novel alpha-helical proteomimetics of the second NR box of SRC2 that exploit structural differences between human estrogen receptor alpha (hERalpha), human estrogen receptor beta (hERbeta), and human thyroid hormone receptor beta (hTRbeta). The resulting library sequentially replaced each leucine with non-natural side chains. Screening this library using a quantitative competition assay revealed compounds that selectively inhibit the interaction of SRC2-2 with each individual NR in preference to its interaction with the other NR. This approach generated highly selective compounds from one that had no specificity for a particular family member. These compounds represent the first family-member-selective competitive inhibitors of the protein interactions of transcription factors. PMID- 12783523 TI - Manipulating supramolecular self-assembly via tailoring pendant group size of linear vinyl polymers. AB - In a series of poly[di(alkyl) vinylterephthalates] (PDAVTs) synthesized via radical polymerization, fine-tuning the size and shape of the side groups manipulated the supramolecular self-assembly and led to control over the formations between amorphous and 2D ordered hexagonal phases. To introduce the 2D long-range ordered structure, the size of the ester side groups at the 2- and 5 positions of the phenyl rings laterally attached to the backbones had to be in the range of propyl/isopropyl to hexyl. The relatively extended backbones observed in these polymers were attributed to steric effects from the side groups. When the n-alkyl groups were larger than hexyl, the ability to form the liquid crystalline phase gradually decreased. A completely disordered phase could be observed by substituting dodecyl groups as side groups. PMID- 12783524 TI - General approach for the synthesis of chiral perylenequinones via catalytic enantioselective oxidative biaryl coupling. AB - By using oxygen as the terminal oxidant, copper complexes derived from chiral 1,5 diaza-cis-decalin catalyze the enantioselective oxidative biaryl coupling of highly functionalized naphthols to provide octa- and decasubstituted binaphthalenes with high selectivity (86-90% ee). Products containing very electron-rich naphthalenes were prone to epimerization under the reaction conditions. This epimerization could be suppressed by employing naphthol starting materials with phenol protecting groups that attenuated the electron-rich nature of the naphthalenes. Direct oxidation of the resultant chiral 1,1'-binaphthol framework completed the first asymmetric synthesis of a perylenequinone containing only an axial chirality element. PMID- 12783525 TI - Detection of short-lived transient protein-protein interactions by intermolecular nuclear paramagnetic relaxation: plastocyanin from Anabaena variabilis. AB - An NMR approach is presented that provides detailed information about short lived, transient interactions between protein molecules in solution. The approach is based on the longitudinal paramagnetic relaxation rates of the protein nuclei and requires that at least one of the interacting molecules is paramagnetic. The specific interactions are monitored by the intermolecular paramagnetic contribution to the relaxation of protons at or close to the interaction surface. By applying the approach to plastocyanin from Anabaena variabilis, specific regions of interaction that may be involved in the electron self-exchange process of this plastocyanin were identified. This is in accord with recent 15N NMR relaxation studies of the backbone dynamics of Anabaena variabilis plastocyanin, with site-directed mutagenesis studies of the functional importance of the corresponding regions in Phormidium laminosum plastocyanin and with the crystal packing surface of P. laminosum plastocyanin. PMID- 12783526 TI - Enantioselective cyclization of racemic supramolecular polymers. AB - Homochiral hydrogen-bonded cyclic assemblies are formed in dilute solutions of racemic supramolecular polymers based on the quadruple hydrogen bonding 2-ureido 4[1H]-pyrimidinone unit, as observed by 1H NMR and SEC experiments. Preorganization of the monomers and the combined binding strength of the eight hydrogen bonds result in a very high stability of the cyclic aggregates with pronounced selectivity between homochiral and heterochiral cyclic species, usually only observed in crystalline or liquid crystalline phases. PMID- 12783527 TI - Photoinduced polarization inversion in a ferroelectric liquid crystal using an ambidextrous chiral thioindigo dopant. AB - In this Communication, we report the first example of photoswitching of a ferroelectric SmC* liquid crystal based on a photoinduced sign inversion of the spontaneous polarization (PS) induced by a single chiral dopant. This is achieved without concomitant destabilization of the SmC* phase using the "ambidextrous" dopant 6-((R,R)-2,3-difluorooct-1-yloxy)-5'-nitro-6'-((R)-2-octyloxy)thioindigo. The (R)-2-octyloxy side chain is sterically coupled to the thioindigo core via the nitro substituent and induces a positive polarization, whereas the (R,R)-2,3 difluorooct-1-yloxy side chain is decoupled from the core and induces a negative polarization. With this new design, the increase in transverse dipole moment of the thioindigo core upon trans-cis photoisomerization raises the polarization power of the coupled 2-octyloxy/thioindigo unit above that of the 2,3 difluorooctyloxy unit and inverts the net sign of PS. PMID- 12783528 TI - Enantioselective total synthesis of ustiloxin D. AB - Ustiloxin D and phomopsin A are potent antimitotic agents that bind to tubulin and interfere with cellular microtubule function. A synthetic strategy has been developed to allow access to both of the natural products as well as a variety of variants of the ustiloxin and phomopsin family members in order to provide sufficient quantities for biological studies. Herein we report the enantioselective total synthesis of ustiloxin D using a longest linear sequence of 20 steps. Four of the five stereocenters were set using catalytic asymmetric methodologies. In particular, Evans's new Al-catalyzed asymmetric aldol reaction facilitated access to both syn and anti products corresponding to the different benzylic stereochemistries found in ustiloxins and phomopsins. In addition, due to its high functional group tolerance, Trost's Pd-mediated etherification was used to construct the chiral tertiary alkyl-aryl ether. Taken together, these synthetic strategies allow us to use densely functionalized intermediates to realize an efficient synthesis of ustiloxin D. PMID- 12783529 TI - Kinetics and thermodynamics of copper(II) binding to apoazurin. AB - The binding of copper(II) to apoazurin has been probed by isothermal titration calorimetry in cholamine buffer at pH 7.0. The standard enthalpy change was determined to be -10.0 +/- 1.4 kcal/mol. Each calorimetric trace reveals an initial exothermic phase followed by an endothermic phase. The calorimetric data could be fit to a kinetic model involving a bimolecular combination of copper(II) and apoazurin in an exothermic process (k = 2 +/-1 x 103 M-1 s-1, DeltaH degrees = -19 +/- 3 kcal/mol) to form an intermediate that spontaneously converts to Cu(II)-azurin in an endothermic process (k = 0.024 +/- 0.01 s-1, DeltaH degrees = +9 +/- 3 kcal/mol). These data suggest that copper(II) first combines with apoazurin in an irreversible process to form an intermediate that converts to copper(II)-azurin in a process driven by the release of water. The overall standard free energy of copper(II) binding to apoazurin is estimated to be -18.8 kcal/mol. PMID- 12783530 TI - Probing the rhodopsin cavity with reduced retinal models at the CASPT2//CASSCF/AMBER level of theory. AB - We show that the ab initio CASPT2//CASSCF strategy previously used to investigate the ground and excited states of the chromophore of the vision receptor rhodopsin (Rh) in vacuo can be successfully implemented in a QM/MM scheme allowing for CASPT2//CASSCF/AMBER geometry optimization and excited state property evaluation in proteins. Two receptor models (Rh-1 and Rh-2) incorporating different reduced chromophores are investigated. It is shown that Rh-2 features a chromophore equilibrium structure with the correct helicity and a lambdamax that is only 52 nm blue-shifted from the observed value. This result should open the way to a qualitatively correct ab initio QM/MM modeling of the early excited state transient species involved in the vision process. PMID- 12783531 TI - Defining space around conducting polymers: reversible protonic doping of a canopied polypyrrole. AB - A canopy-shaped pyrrole derivative 2 was prepared, in which a sterically demanding pendant group is juxtaposed to the pyrrole fragment to minimize interstrand pi-pi stacking interactions in the resulting polymer. Anodic polymerization of 2 afforded highly conductive poly(2), the electronic structure of which was probed by various spectroelectrochemical techniques. A limited charge delocalization within poly(2) translates into a well-defined conductivity profile, properties important for resistivity-based sensing. Notably, the bulk conductivity was precisely modulated by a rapid and reversible deprotonation and reprotonation of the polymer backbone. PMID- 12783532 TI - Formation and properties of cyclo[6]pyrrole and cyclo[7]pyrrole. AB - Oxidative coupling of a tetraalkylbipyrrole under FeIII-mediated coupling conditions in the presence of HCl results in a mixture of cyclo[6]- and cyclo[7]pyrroles, as well as the known cyclo[8]pyrrole. This "matched set" of heteroannulenes was analyzed by spectroscopic, electrochemical, and X-ray diffraction methods. PMID- 12783533 TI - Cytosol-mimetic chemistry: kinetics of the trypsin-catalyzed hydrolysis of p nitrophenyl acetate upon addition of polyethylene glycol and N-tert-butyl acetoacetamide. AB - The sensitivity of an enzyme to its environment has provoked much interest both for its immediate relevance to biochemistry and for the use of enzymes in chemical synthesis. The intercellular or extracellular environment in which an enzyme naturally operates is crowded with macromolecular, small-molecule, and ionic solutes and hence is markedly different from the dilute aqueous buffer solutions commonly cited for comparisons of biochemical processes. We report the results of a kinetic study into the effects of such a crowded solution on the rate of an enzyme-mediated process-the trypsin-catalyzed hydrolysis of a nonnatural substrate ester. The catalytic rate constant decreases linearly with solvent polarity, but substrate binding is independent of the concentration of added crowding agent up to 395 g/L. PMID- 12783534 TI - A gate to organokrypton chemistry: HKrCCH. AB - An organic molecule containing krypton, HKrCCH, is reported. The preparation of HKrCCH includes 193-nm photolysis of H2C2/Kr solid mixtures at 8 K and subsequent thermal mobilization of hydrogen atoms at >/=30 K. The identification is based on infrared absorption spectroscopy and supported by ab initio calculations which show ionic and covalent contributions to the bonding. We believe that a series of similar organokrypton molecules can be prepared as computationally demonstrated for HKrC4H and HKrC3H3. These results feature a generally novel way for activating chemically the H-CC- group, which can find practical applications of the krypton catalysis. PMID- 12783535 TI - Novel binding and efficient cellular uptake of guanidine-based peptide nucleic acids (GPNA). AB - Incorporation of a guanidine functional group into the PNA backbone facilitates cellular uptake of PNA into mammalian cells with efficiency comparable to that of the TAT transduction domain. The modified PNA recognizes and binds to the complementary DNA strand in accordance with Watson-Crick recognition rules. However, unlike polypyrimidine PNA which binds to DNA in 2:1 stoichiometry, the modified PNA binds to complementary DNA in a 1:1 ratio to form a highly stable duplex. PMID- 12783536 TI - Deoxyribozymes that synthesize branched and lariat RNA. AB - Branched RNA molecules with a 2',5'-phosphodiester linkage are important biochemical intermediates. Lariat RNA is a particular type of branched RNA that is formed during intron splicing in vivo. Synthesis of branched and lariat RNA is challenging, and there are few general approaches that are applicable in vitro. Here we report the identification of divalent metal-dependent deoxyribozymes (DNA enzymes) that synthesize branched and lariat RNA. In vitro selection was used to obtain deoxyribozymes that selectively join an internal RNA 2'-hydroxyl with a 5' terminal triphosphate in a convenient "binding arms" format. At least 85% yield of 2',5'-branched RNA is obtained at 37 degrees C and 20 mM Mn2+, pH 7.5 in 1) at high temperatures. The temperature dependence of the EIE for oxidative addition of CH4 and CD4 differs significantly from that for coordination, with the EIE being normal at all temperatures and approaching infinity at 0 K. In contrast to oxidative addition of methane which is normal at all temperatures, the EIE for oxidative addition of H2 and D2 exhibits a transition from inverse to normal upon raising the temperature. The existence of inverse EIEs in these systems at low temperatures is a result of the zero point energy changes for the products upon isotopic substitution being greater than those for the reactants (H2 or CH4). PMID- 12783539 TI - Chrysanthemyl diphosphate synthase. The relationship among chain elongation, branching, and cyclopropanation reactions in the isoprenoid biosynthetic pathway. AB - The genes for chrysanthemyl diphosphate (CPP) synthase and farnesyl diphosphate (FPP) synthase from sagebrush, Artemisia tridentata spiciformis, were used to prepare a series of chimeric proteins to investigate the 1'-4 chain elongation, 1'-2 branching, and c1'-2-3 cyclopropanation reactions that join isoprenoid units to build more complex structures. The two genes were modified by site-directed mutagenesis to generate an identical set of six unique restriction sites at identical locations. The locations were selected to place a restriction site between each of the five conserved regions found in prenyltransferases that catalyze chain elongation. A series of chimeric proteins were generated by replacing amino acids in FPP synthase, beginning at the N-terminus of the enzyme, with increasing stretches of peptide from CPP synthase. An analysis of the products produced by the chimeras revealed a transition from 1'-4 chain elongation, to 1'-2 branching, and ultimately to c1'-2-3 cyclopropanation. These results demonstrate that the catalytic site for chain elongation, with minor modifications in its architecture, also catalyzes 1'-2 branching and c1'-2-3 cyclopropanation, and suggest that the branching and cyclopropanation reactions, in analogy to chain elongation, are electrophilic alkylations. PMID- 12783541 TI - Contributions of conformational compression and preferential transition state stabilization to the rate enhancement by chorismate mutase. AB - The rate enhancement provided by the chorismate mutase (CM) enzyme for the Claisen rearrangement of chorismate to prephenate has been investigated by application of the concept of near attack conformations (NACs). Using a combined QM/MM Monte Carlo/free-energy perturbation (MC/FEP) method, 82% and 100% of chorismate conformers were found to be NAC structures in water and in the CM active site, respectively. Consequently, the conversion of non-NACs to NACs does not contribute to the free energy of activation from preorganization of the substrate into NACs. The FEP calculations yielded differences in free energies of activation that well reproduce the experimental data. Additional calculations indicate that the rate enhancement by CM over the aqueous phase results primarily from conformational compression of NACs by the enzyme and that this process is enthalpically controlled. This suggests that preferential stabilization of the transition state in the enzyme environment relative to water plays a secondary role in the catalysis by CM. PMID- 12783542 TI - Incorporation of trifluoroisoleucine into proteins in vivo. AB - Two fluorinated derivatives of isoleucine: d,l-2-amino-3-trifluoromethyl pentanoic acid (3TFI, 2) and d,l-2-amino-5,5,5-trifluoro-3-methyl pentanoic acid (5TFI, 3) were prepared. 5TFI was incorporated into a model target protein, murine dihydrofolate reductase (mDHFR), in an isoleucine auxotrophic Escherichia coli host strain suspended in 5TFI-supplemented minimal medium depleted of isoleucine. Incorporation of 5TFI was confirmed by tryptic peptide analysis and matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) of the protein product. Amino acid analysis showed that more than 93% of the encoded isoleucine residues were replaced by 5TFI. Measurement of the rate of activation of 5TFI by the E. coli isoleucyl-tRNA synthetase (IleRS) yielded a specificity constant (k(cat)/K(m)) 134-fold lower than that for isoleucine. 5TFI was successfully introduced into the cytokine murine interleukin-2 (mIL-2) at the encoded isoleucine positions. The concentration of fluorinated protein that elicits 50% of the maximal proliferative response is 3.87 ng/mL, about 30% higher than that of wild-type mIL-2 (EC(50) = 2.70 ng/mL). The maximal responses are equivalent for the fluorinated and wild-type cytokines, indicating that fluorinated proteins can fold into stable and functional structures. 3TFI yielded no evidence for in vivo incorporation into recombinant proteins, and no evidence for activation by IleRS in vitro. PMID- 12783543 TI - Shape-persistant macrocycles with terpyridine units: synthesis, characterization, and structure in the crystal. AB - The synthesis of a variety of shape-persistent macrocycles with either one (1a-d, 2) or two (opposing) terpyridine units (3, 4, 5a-c) and inner diameters of up to 2 nm is described. The sequences are mainly based on transition metal cross coupling reactions and, whenever appropriate, compared with one another regarding their respective efficiency. Typical overall yields and amounts prepared range from 8% (4) to 27% (3) and 25 mg (1a) to 290 mg (1b), respectively. For solubility and processing of the targeted cycles, all precursors have already been decorated with flexible side chains (hexyloxy or hexyloxymethyl). The cycles' characterization is based on MALDI-TOF mass spectrometry, 2D NMR spectroscopy, and/or low-temperature single-crystal X-ray diffraction. Their packing in the crystal is discussed in terms of both number and length of side chains. Cycle 1d was physisorbed into an ordered structure at the solution-HOPG interface and investigated by scanning tunneling microscopy (STM). PMID- 12783544 TI - Light-driven decarboxylation of wild-type green fluorescent protein. AB - The response of wild-type GFP to UV and visible light was investigated using steady state absorption, fluorescence, and Raman spectroscopies. As reported previously [van Thor, Nat. Struct. Biol. 2002, 9, 37-41], irradiation of GFP results in decarboxylation of E222. Here it is reported that the rate of the light-driven decarboxylation reaction strongly depends on the excitation wavelength, decreasing in the order 254 nm > 280 nm > 476 nm. The relative efficiencies of decarboxylation are explained in terms of the Kolbe-type mechanism in which the excited state of the chromophore acts as an oxidant by accepting an electron from E222. Specifically, it is proposed that 254 nm excitation populates the S2 (or higher) excited state of the chromophore, whereas 404 and 476 nm excitation populate the S1 excited state of neutral and anionic forms, respectively, and that the relative oxidizing power of the three excited states controls the rate of the decarboxylation reaction. In addition, the role of W57 in the photophysics of GFP has been probed by mutating this residue to phenylalanine. These studies reveal that while W57 does not affect decarboxylation, this residue is involved in resonance energy transfer with the chromophore, thereby partially explaining the green fluorescence observed upon UV irradiation of wild-type GFP. Finally, comparison of Raman spectra obtained from nonilluminated and decarboxylated forms of wild-type GFP has provided further vibrational band assignments for neutral and anionic forms of the chromophore within the protein. In addition, these spectra provide valuable insight into the specific interactions between the protein and the chromophore that control the optical properties of wild-type GFP. PMID- 12783545 TI - Direct determination of the complete set of iron normal modes in a porphyrin imidazole model for carbonmonoxy-heme proteins: [Fe(TPP)(CO)(1-MeIm)]. AB - Detailed Fe vibrational spectra have been obtained for the heme model complex [Fe(TPP)(CO)(1-MeIm)] using a new, highly selective and quantitative technique, Nuclear Resonance Vibrational Spectroscopy (NRVS). This spectroscopy measures the complete vibrational density of states for iron atoms, from which normal modes can be calculated via refinement of the force constants. These data and mode assignments can reveal previously undetected vibrations and are useful for validating predictions based on optical spectroscopies and density functional theory, for example. Vibrational modes of the iron porphyrin-imidazole compound [Fe(TPP)(CO)(1-MeIm)] have been determined by refining normal mode calculations to NRVS data obtained at an X-ray synchrotron source. Iron dynamics of this compound, which serves as a useful model for the active site in the six coordinate heme protein, carbonmonoxy-myoglobin, are discussed in relation to recently determined dynamics of a five-coordinate deoxy-myoglobin model, [Fe(TPP)(2-MeHIm)]. For the first time in a six-coordinate heme system, the iron imidazole stretch mode has been observed, at 226 cm(-)(1). The heme in-plane modes with large contributions from the nu(42), nu(49), nu(50), and nu(53) modes of the core porphyrin are identified. In general, the iron modes can be attributed to coupling with the porphyrin core, the CO ligand, the imidazole ring, and/or the phenyl rings. Other significant findings are the observation that the porphyrin ring peripheral substituents are strongly coupled to the iron doming mode and that the Fe-C-O tilting and bending modes are related by a negative interaction force constant. PMID- 12783546 TI - Approaching real-time molecular diagnostics: single-pair fluorescence resonance energy transfer (spFRET) detection for the analysis of low abundant point mutations in K-ras oncogenes. AB - The aim of this study was to develop new strategies for analyzing molecular signatures of disease states approaching real-time using single pair fluorescence resonance energy transfer (spFRET) to rapidly detect point mutations in unamplified genomic DNA. In addition, the detection process was required to discriminate between normal and mutant (minority) DNAs in heterogeneous populations. The discrimination was carried out using allele-specific primers, which flanked the point mutation in the target gene and were ligated using a thermostable ligase enzyme only when the genomic DNA carried this mutation. The allele-specific primers also carried complementary stem structures with end labels (donor/acceptor fluorescent dyes, Cy5/Cy5.5, respectively), which formed a molecular beacon following ligation. We coupled ligase detection reaction (LDR) with spFRET to identify a single base mutation in codon 12 of a K-ras oncogene that has high diagnostic value for colorectal cancers. A simple diode laser-based fluorescence system capable of interrogating single fluorescent molecules undergoing FRET was used to detect photon bursts generated from the molecular beacon probes formed upon ligation. LDR-spFRET provided the necessary specificity and sensitivity to detect single-point mutations in as little as 600 copies of human genomic DNA directly without PCR at a level of 1 mutant per 1000 wild type sequences using 20 LDR thermal cycles. We also demonstrate the ability to rapidly discriminate single base differences in the K-ras gene in less than 5 min at a frequency of 1 mutant DNA per 10 normals using only a single LDR thermal cycle of genomic DNA (600 copies). Real-time LDR-spFRET detection of point mutations in the K-ras gene was accomplished in PMMA microfluidic devices using sheath flows. PMID- 12783547 TI - EPR spin-trapping evidence for the direct, one-electron reduction of tert butylhydroperoxide to the tert-butoxyl radical by copper(II): paradigm for a previously overlooked reaction in the initiation of lipid peroxidation. AB - Lipid peroxidation is often initiated using Cu(II) ions. It is widely assumed that Cu(II) oxidizes preformed lipid hydroperoxides to peroxyl radicals, which propagate oxidation of the parent fatty acid via hydrogen atom abstraction. However, the oxidation of alkyl hydroperoxides by Cu(II) is thermodynamically unfavorable. An alternative means by which Cu(II) ions could initiate lipid peroxidation is by their one-electron reduction of lipid hydroperoxides to alkoxyl radicals, which would be accompanied by the generation of Cu(III). We have investigated by EPR spectroscopy, in conjunction with the spin trap 5,5 dimethyl-1-pyrroline N-oxide, the reactions of various Cu(II) chelates with tert butylhydroperoxide. Spectra contained signals from the tert-butoxyl, methyl, and methoxyl radical adducts. In many previous studies, the signal from the methoxyl adduct has been assigned incorrectly to the tert-butylperoxyl adduct, which is now known to be unstable, releasing the tert-butoxyl radical upon decomposition. This either is trapped by 5,5-dimethyl-1-pyrroline N-oxide or undergoes beta scission to the methyl radical, which either is trapped or reacts with molecular oxygen to give, ultimately, the methoxyl radical adduct. By using metal chelates that are known to be specific in either their oxidation or reduction of tert butylhydroperoxide (the Cu(II) complex of bathocuproine disulfonic acid and the Fe(II) complex of diethylenetriaminepentaacetic acid, respectively) for comparison, we have been able to deduce, from the relative concentrations of the three radical adducts, that the Cu(II) complexes tested each reduce tert butylhydroperoxide directly to the tert-butoxyl radical. These findings suggest that a previously overlooked reaction, namely the direct reduction of preformed lipid hydroperoxides to alkoxyl radicals by Cu(II), may be responsible for the initiation of lipid peroxidation by Cu(II) ions. PMID- 12783548 TI - Characterization of supramolecular (H2O)18 water morphology and water-methanol (H2O)15(CH3OH)3 clusters in a novel phosphorus functionalized trimeric amino acid host. AB - Phosphorus functionalized trimeric alanine compounds (l)- and (d) P(CH(2)NHCH(CH(3))COOH)(3) 2 are prepared in 90% yields by the Mannich reaction of Tris(hydroxymethyl)phosphine 1 with (l)- or (d)- Alanine in aqueous media. The hydration properties of (l)-2 and (d)-2 in water and water-methanol mixtures are described. The crystal structure analysis of (l)-2.4H(2)O, reveals that the alanine molecules pack to form two-dimensional bilayers running parallel to (001). The layered structural motif depicts two closely packed monolayers of 2 each oriented with its phosphorus atoms projected at the center of the bilayer and adjacent monolayers are held together by hydrogen bonds between amine and carboxylate groups. The water bilayers are juxtaposed with the H-bonded alanine trimers leading to 18-membered (H(2)O)(18) water rings. Exposure of aqueous solution of (l)-2 and (d)-2 to methanol vapors resulted in closely packed (l)-2 and (d)-2 solvated with mixed water-methanol (H(2)O)(15)(CH(3)OH)(3) clusters. The O-O distances in the mixed methanol-water clusters of (l)-2.3H(2)O.CH(3)OH and (d)-2.3H(2)O.CH(3)OH (O-O(average) = 2.857 A) are nearly identical to the O-O distance observed in the supramolecular (H(2)O)(18) water structure (O-O(average) = 2.859 A) implying the retention of the hydrogen bonded structure in water despite the accommodation of hydrophobic methanol groups within the supramolecular (H(2)O)(15)(CH(3)OH)(3) framework. The O-O distances in (l) 2.3H(2)O.CH(3)OH and (d)-2.3H(2)O.CH(3)OH and in (H(2)O)(18) are very close to the O-O distance reported for liquid water (2.85 A). PMID- 12783549 TI - Theoretical study of the amazing firefly bioluminescence: the formation and structures of the light emitters. AB - Reaction mechanisms for the formation of the keto-form of oxyluciferin (OxyLH(2)) from the luciferin of fireflies via a dioxetanone intermediate are predicted using the B3LYP/6-31G theoretical method. The ring opening of a model dioxetanone and the decarboxylation proceed in one step via a singlet diradical transition structure with an activation barrier of 18.1 and an exothermicity of 90.8 kcal/mol. The S(0) --> S(1) vertical excitation energies predicted with time dependent density functional theory, TDDFT B3LYP/6-31+G, for the anionic and neutral forms of OxyLH(2) are in the range of 60 to 80 kcal/mol. These energetic results support the generally accepted theory of chemically initiated electron exchange luminescence (CIEEL). The chemical origin of the multicolor bioluminescence from OxyLH(2) is examined theoretically using the TDDFT B3LYP/6 31+G, ZINDO//B3LYP/6-31+G, and CIS/6-31G methods. A change in color of the light emission upon rotation of the two rings in the S(1) excited state of OxyLH(2) is unlikely because both possible emitters, the planar keto- and enol-forms, are minima on the S(1) potential energy surface. The participation of the enol-forms of OxyLH(2) in bioluminescence is plausible but not required to explain the multicolor emission. According to predictions at the TDDFT B3LYP level, the color of the bioluminescence depends on the polarization of the OxyLH(2) in the microenvironment of the enzyme-OxyLH(2) complex. PMID- 12783550 TI - Nitroxyl disulfides, novel intermediates in transnitrosation reactions. AB - A novel anionic RSN(O)SR species, the intermediate in transnitrosation reactions, was explored computationally with B3LYP and CBS-QB3 methods. The species resembles a nitroxyl coordinated to a highly distorted disulfide, and it differs significantly from intermediates in nucleophilic acyl substitution. Reactions of the following species were computed for comparison: MeS(-) + MeSNO; MeO(-) + MeONO; MeS(-) + MeSCHO; MeO(-) + MeOCHO. The last two have very different intermediates from the first two. Mass spectrometric experimental evidence is presented that is consistent with the formation of a nitroxyl disulfide in the gas phase. The calculated proton affinity and redox potentials of the intermediate are also reported. PMID- 12783551 TI - Fluorescence resonance energy transfer (FRET) as a high-throughput assay for coupling reactions. Arylation of amines as a case study. AB - A solution-phase assay based on fluorescence resonance energy transfer (FRET) was developed for high-throughput screening of palladium catalyzed aminations of aryl halides. Dansylpiperazine was used as the fluorescent component and a chloro- or bromoarene tagged with an azodye as the quenching partner. Fluorescence intensities of reaction aliquots correlated linearly with reaction yield after dilution to appropriate concentrations. A library of 119 phosphine and heterocyclic carbene ligands was evaluated in duplicate reactions of two combinations. In general, the FRET assay displayed excellent reproducibility, with less than 5% of the duplicate experiments showing significant variability in yields. Among reactions producing greater than 50% yield, the average percent uncertainty was just 5%. For a small subset of sterically hindered ligands, differences in yields between 10 and 20% were observed between the substrates bearing dyes for the FRET assay and substrates that are unfunctionalized. However, the remaining catalyst combinations gave yields similar to those expected from literature precedent. In addition to an evaluation of the accuracy of the FRET assay, this work includes the use of the FRET assay to investigate relative activities of various catalysts for the amination of aryl bromides and chlorides and to find conditions for aminations in more polar solvents. Reactions with K(3)PO(4) base in aqueous mixtures of polar and nonpolar organic solvents were shown to be appropriate for the amination chemistry. PMID- 12783552 TI - Stereoblock copolymers and tacticity control in controlled/living radical polymerization. AB - Three controlled/living radical polymerization processes, atom transfer radical polymerization (ATRP), reversible addition-fragmentation transfer (RAFT) polymerization, and nitroxide-mediated polymerization (NMP), were investigated for the polymerization of N,N-dimethylacrylamide in the presence of Lewis acids known to enhance isotacticity, such as yttrium trifluoromethanesulfonate (Y(OTf)(3)) and ytterbium trifluoromethanesulfonate (Yb(OTf)(3)). Poly(N,N dimethylacrylamide) with controlled molecular weight, low polydispersity (M(w)/M(n) < 1.2), and a high proportion of meso dyads ( approximately 85%) was prepared by ATRP (with initiating system methyl 2 chloropropionate/CuCl/Me(6)TREN) and RAFT (with cumyl dithiobenzoate transfer agent) in the presence of Y(OTf)(3). The combination of NMP (using N-tert-butyl-1 diethylphosphono-2,2-dimethylpropyl nitroxide, SG1) and a Lewis acid complexation technique led to less precise control over chain architecture and microstructure ( approximately 65% meso dyads), as compared to RAFT/Y(OTf)(3) or ATRP/Y(OTf)(3). The latter two systems were used for the first one-pot synthesis of stereoblock copolymers by radical polymerization. Well-defined stereoblock copolymers, atactic-b-isotactic poly(N,N-dimethylacrylamides), were obtained by adding Y(OTf)(3) at a given time to either RAFT or ATRP polymerizations, initially started without the presence of the Lewis acid. PMID- 12783553 TI - Modeling the reaction mechanisms of the amide hydrolysis in an N-(o carboxybenzoyl)-L-amino acid. AB - Reaction mechanisms of the amide hydrolysis from the protonated, neutral, and deprotonated forms of N-(o-carboxybenzoyl)-l-amino acid have been investigated by use of the B3LYP density functional method. Our calculations reveal that in the amide hydrolysis the reaction barrier is significantly lower in solution than that in the gas phase, in contrast with the mechanism for imide formation in which the solvent has little influence on the reaction barrier. In the model reactions, the water molecules function both as a catalyst and as a reactant. The reaction mechanism starting from the neutral form of N-(o-carboxybenzoyl)-l-amino acid, which corresponds to pH 0-3, is concluded to be the most favored, and a concerted mechanism is more favorable than a stepwise mechanism. This conclusion is in agreement with experimental observations that the optimal pH range for amide hydrolysis of N-(o-carboxybenzoyl)-l-leucine is pH 0-3 where N-(o carboxybenzoyl)-l-leucine is predominantly in its neutral form. We suggest that besides the acid-catalyzed mechanism the addition-elimination mechanism is likely to be an alternative choice for cleaving an amide bond. For the reaction mechanism initiated by protonation at the amidic oxygen (hydrogen ion concentration H(0) < -1), the reaction of the model compound with two water molecules lowers the transition barrier significantly compared with that involving a single water molecule. PMID- 12783554 TI - Ion pairing and host-guest complexation in low dielectric constant solvents. AB - We report an equilibrium treatment for complexation of ionic species in low dielectric constant media that explicitly includes ion pairing of one of the components. Experimental validation was achieved through study of pseudorotaxane formation between dibenzylammonium salts and dibenzo-24-crown-8. In particular, we show that concentration-dependent fluctuations in the apparent K(a,exp) values as usually reported are attributable to ion pairing, with dissociation constant K(ipd), and that the constant K(ap) for complexation of the free cationic guest species, G(+), by the host crown ether is independent of counterion. More generally, using a simple extension of our model, we show the ability to diagnose the relative extent of ion pairing of the complex, which may be readily applied to other host-guest systems involving ionic species. PMID- 12783555 TI - Mechanistic investigations of the palladium-catalyzed aerobic oxidative kinetic resolution of secondary alcohols using (-)-sparteine. AB - The mechanistic details of the Pd(II)/(-)-sparteine-catalyzed aerobic oxidative kinetic resolution of secondary alcohols were elucidated, and the origin of asymmetric induction was determined. Saturation kinetics were observed for rate dependence on [(-)-sparteine]. First-order rate dependencies were observed for both the Pd((-)-sparteine)Cl(2) concentration and the alcohol concentration at high and low [(-)-sparteine]. The oxidation rate was inhibited by addition of (-) sparteine HCl. At low [(-)-sparteine], Pd-alkoxide formation is proposed to be rate limiting, while at high [(-)-sparteine], beta-hydride elimination is proposed to be rate determining. These conclusions are consistent with the measured kinetic isotope effect of k(H)/k(D) = 1.31 +/- 0.04 and a Hammett rho value of -1.41 +/- 0.15 at high [(-)-sparteine]. Calculated activation parameters agree with the change in the rate-limiting step by increasing [(-)-sparteine] with DeltaH(++) = 11.55 +/- 0.65 kcal/mol, DeltaS(++) = -24.5 +/- 2.0 eu at low [(-)-sparteine], and DeltaH(++) = 20.25 +/- 0.89 kcal/mol, DeltaS() = -5.4 +/- 2.7 eu at high [(-)-sparteine]. At high [(-)-sparteine], the selectivity is influenced by both a thermodynamic difference in the stability of the diastereomeric Pd-alkoxides formed and a kinetic beta-hydride elimination to maximize asymmetric induction. At low [(-)-sparteine], the selectivity is influenced by kinetic deprotonation, resulting in lower k(rel) values. A key, nonintuitive discovery is that (-)-sparteine plays a dual role in this oxidative kinetic resolution of secondary alcohols as a chiral ligand on palladium and as an exogenous chiral base. PMID- 12783556 TI - Metal ion-promoted intramolecular electron transfer in a ferrocene-naphthoquinone linked dyad. Continuous change in driving force and reorganization energy with metal ion concentration. AB - Thermal intramolecular electron transfer from the ferrocene (Fc) to naphthoquinone (NQ) moiety occurs efficiently by the addition of metal triflates (M(n)()(+): Sc(OTf)(3), Y(OTf)(3), Eu(OTf)(3)) to an acetonitrile solution of a ferrocene-naphthoquinone (Fc-NQ) linked dyad with a flexible methylene and an amide spacer, although no electron transfer takes place in the absence of M(n)()(+). The resulting semiquinone radical anion (NQ(*)(-)) is stabilized by the strong binding of M(n)()(+) with one carbonyl oxygen of NQ(*)(-)( )()as well as hydrogen bonding between the amide proton and the other carbonyl oxygen of NQ(*)(-). The high stability of the Fc(+)()-NQ(*)(-)/M(n)()(+)() complex allows us to determine the driving force of electron transfer by the conventional electrochemical method. The one-electron reduction potential of the NQ moiety of Fc-NQ is shifted to a positive direction with increasing concentration of M(n)()(+), obeying the Nernst equation, whereas the one-electron oxidation potential of the Fc moiety remains the same. The driving force dependence of the observed rate constant (k(ET)) of M(n)()(+)-promoted intramolecular electron transfer is well evaluated in light of the Marcus theory of electron transfer. The driving force of electron transfer increases with increasing concentration of M(n)()(+) [M(n)()(+)], whereas the reorganization energy of electron transfer decreases with increasing [M(n)()(+)] from a large value which results from the strong binding between NQ(*)(-) and M(n)()(+). PMID- 12783557 TI - MLCT state structure and dynamics of a copper(I) diimine complex characterized by pump-probe X-ray and laser spectroscopies and DFT calculations. AB - The molecular structure and dynamics of the photoexcited metal-to-ligand-charge transfer (MLCT) state of [Cu(I)(dmp)(2)](+), where dmp is 2,9-dimethyl-1,10 phenanthroline, in acetonitrile have been investigated by time-domain pump-probe X-ray absorption spectroscopy, femtosecond optical transient spectroscopy, and density functional theory (DFT). The time resolution for the excited state structural determination was 100 ps, provided by single X-ray pulses from a third generation synchrotron source. The copper ion in the thermally equilibrated MLCT state has the same oxidation state as the corresponding copper(II) complex in the ground state and was found to be penta-coordinate with an average nearest neighbor Cu-N distance 0.04 A shorter than that of the ground state [Cu(I)(dmp)(2)](+). The results confirm the previously proposed "exciplex" structure of the MLCT state in Lewis basic solvents. The evolution from the photoexcited Franck-Condon MLCT state to the thermally equilibrated MLCT state was followed by femtosecond optical transient spectroscopy, revealing three time constants of 500-700 fs, 10-20 ps, and 1.6-1.7 ns, likely related to the kinetics for the formation of the triplet MLCT state, structural relaxation, and the MLCT excited-state decay to the ground state, respectively. DFT calculations are used to interpret the spectral shift on structural relaxation and to predict the geometries of the ground state, the tetracoordinate excited state, and the exciplex. The DFT calculations also indicate that the amount of charge transferred from copper to the dmp ligand upon photoexcitation is similar to the charge difference at the copper center between the ground-state copper(I) and copper(II) complexes. PMID- 12783558 TI - Intermolecular activation of hydrocarbon C-H bonds under ambient conditions by 16 electron neopentylidene and benzyne complexes of molybdenum. AB - CpMo(NO)(CH(2)CMe(3))(2) (1), a complex with alpha-agostic C-H.Mo interactions, evolves neopentane in neat hydrocarbon solutions at room temperature and forms the transient 16-electron alkylidene complex, CpMo(NO)(=CHCMe(3)), which subsequently activates solvent C-H bonds. Thus, it reacts with tetramethylsilane or mesitylene to form CpMo(NO)(CH(2)CMe(3))(CH(2)SiMe(3)) (2) or CpMo(NO)(CH(2)CMe(3))(eta(2)-CH(2)C(6)H(3)-3,5-Me(2)) (3), respectively, in nearly quantitative yields. Under identical conditions, 1 in p-xylene generates a mixture of sp(2) and sp(3) C-H bond activation products, namely CpMo(NO)(CH(2)CMe(3))(C(6)H(3)-2,5-Me(2)) (4, 73%) and CpMo(NO)(CH(2)CMe(3))(eta(2)-CH(2)C(6)H(4)-4-Me) (5, 27%). In benzene at room temperature, 1 transforms to a mixture of CpMo(NO)(CH(2)CMe(3))(C(6)H(5)) (6) and CpMo(NO)(C(6)H(5))(2) (7) in a sequential manner. Most interestingly, the thermal activation of 6 at ambient temperatures gives rise to two parallel modes of reactivity involving either the elimination of benzene and formation of CpMo(NO)(=CHCMe(3)) or the elimination of neopentane and formation of the benzyne complex, CpMo(NO)(eta(2)-C(6)H(4)). In pyridine, these intermediates are trapped as the isolable 18-electron adducts, CpMo(NO)(=CHCMe(3))(NC(5)H(5)) (8) and CpMo(NO)(eta(2)-C(6)H(4))(NC(5)H(5)) (9), and, in hydrocarbon solvents, they effect the intermolecular activation of aliphatic C-H bonds at room temperature to generate mixtures of neopentyl- and phenyl-containing derivatives. However, the distribution of products resulting from the hydrocarbon activations is dependent on the nature of the solvent, probably due to solvation effects and the presence of sigma- or pi-hydrocarbon complexes on the reaction coordinates of the alkylidene and the benzyne intermediates. The results of DFT calculations on these processes in the gas phase support the existence of such hydrocarbon complexes and indicate that better agreement with experimental observations is obtained when the actual neopentyl ligand rather than the simpler methyl ligand is used in the model complexes. PMID- 12783559 TI - From 1D chain to 3D network: tuning hybrid II-VI nanostructures and their optical properties. AB - In an effort to make semiconductor nanomaterials with tunable properties, we have deliberately designed and synthesized a family of novel organic-inorganic hybrid nanocomposites based on II-VI semiconductors with structures ranging from one dimensional (1-D) chain to two-dimensional layer (2-D) to three-dimensional (3-D) framework. All nanostructures exhibit strong quantum confinement effect (QCE), while possessing a perfectly periodic arrangement. The optical absorption experiments show that all compounds generate a very large blue shift in the absorption edge (1.0-2.0 eV) due to the strong QCE. More significantly, their band edge shift and optical properties can be tuned by changing the dimensionality of inorganic motifs as well as overall crystal structures. Raman studies reveal that not only do these structures have distinctly different vibrational signatures from those of the II-VI host semiconductors, but they also differ significantly from each other as a result of changes in dimensionality. The crystal structures of these nanocomposite materials have been characterized by single crystal and/or powder X-ray diffraction methods. [ZnTe(pda)] (1; pda = propanediamine) is composed of 1-D chains of [ZnTe] with pda chelating to Zn atoms. [ZnTe(N(2)H(4))] (2; N(2)H(4) = hydrazine) and [ZnTe(ma)] (3; ma = MeNH(2) = methylamine) are two-dimensional (2-D) layered structures containing [ZnTe] slabs and terminal hydrazine (2) or methylamine (3) molecules. The crystal structures of [CdSe(en)(0.5)] (4; en = ethylenediamine) and [CdSe(pda)(0.5)] (5) are 3-D networks containing [CdSe] slabs bridged by bidentate organic diamine molecules. Crystal data for 1: Orthorhombic, space group Pbcm, a = 9.997(2), b = 6.997(1), c = 10.332(2) A, Z = 4. For 2: Monoclinic, space group P2(1), a = 4.2222(6), b = 6.9057(9), c = 7.3031(10) A, beta = 98.92(8) degrees, Z = 2. For 3: Orthorhombic, Pbca, a = 7.179(1), b = 6.946(1), c = 18.913(4) A, Z = 8. For 4: Orthorhombic, Pbca, a = 7.0949(3), b = 6.795(3), c = 16.7212(8) A, Z = 8. For 5: Orthorhombic, Cmc2(1), a = 20.6660(12), b = 6.8900(4), c = 6.7513(4) A, Z = 8. PMID- 12783560 TI - Substrate binding to NO-ferro-naphthalene 1,2-dioxygenase studied by high resolution Q-band pulsed 2H-ENDOR spectroscopy. AB - The active site of naphthalene 1,2-dioxygenase (NDO) contains a Rieske Fe-S cluster and a mononuclear non-heme iron, which are contributed by different alpha subunits in the (alphabeta)(3) structure. The enzyme catalyzes cis dihydroxylation of aromatic substrates, in addition to numerous other adventitious oxidation reactions. High-resolution Mims (2)H-ENDOR (electron nuclear double resonance) spectra have been recorded for the NO-ferrous center of NDO bound with the substrates d(8)-naphthalene, d(2)-naphthalene, d(8)-toluene, d(3)-toluene, and d(6)-benzene; samples were prepared in a D(2)O buffer to test for solvent-derived ligands; spectra were collected for enzymes with the Rieske diiron center in both its oxidized and reduced states. A sharp quartet ENDOR pattern from a nearby deuteron of the substrate in a major binding geometry (denoted as A) was detected for all perdeuterated substrates. Examination of the sample prepared with 1,4-di-deutero-naphthalene shows that the signal arises from D1. Analysis of two-dimensional (2-D) orientation-selective ENDOR patterns collected for this sample defined the location of the D1 deuteron, with respect to the g-frame of the iron center and the orientation of the C-D1 bond. Consideration of the orientations of naphthalene that are permitted within the constraints of these results, as supported by a novel approach to simulations of orientation-selective, 2-D ENDOR patterns for the perdeuterated naphthalene sample, which summed contributions from D1/D2/D8, disclose the geometry of the naphthalene and the Fe-NO fragment. The two deuterons of the reactive carbons, D1 and D2, are closest to the Fe atom (r(Fe)(-)(D1) approximately 4.3 A, r(Fe)( )(D2) approximately 5.0 A), whereas D8 is farther away (r(Fe)(-)(D8) approximately 5.3 A). Perhaps more instructive, D1-N and D2-N distances to the O(2) surrogate, NO, are approximately 2.4 and approximately 3.3 A, respectively, whereas the D8-N distance is approximately 3.7 A. The data show that benzene and the aromatic ring of toluene also sit within the substrate-binding pocket adjacent to the mononuclear Fe atom. These rings occupy a position similar to that of the "proximal" ring of naphthalene, with the closest ring deuteron being located at a distance of approximately 4.3-4.4 A from the Fe atom and with the Fe D vector being slightly off the Fe-N(O) direction. In particular, comparison of the data for d(8)-toluene and methyl-d(3)-toluene shows that the methyl group of toluene points away from the Fe atom, despite observations that the oxidation of toluene occurs at the methyl group during catalysis. The Rieske cluster is reduced during both steady-state and single-turnover catalysis; therefore, the effect of its oxidation state on the geometry of substrate binding was examined. The spectra from the NDO-naphthalene complex also revealed a second binding conformation (denoted as B), in which the substrate is located approximately 0.5 A farther from the Fe atom. The relative populations of A- and B-sites are allosterically changed when the Rieske cluster is reduced. ENDOR of exchangeable protons shows that the water/hydroxide of Fe-NDO is retained upon binding NO. PMID- 12783561 TI - Spectroscopy and electronic structure of electron deficient zinc phthalocyanines. AB - The effect of introduction of perfluoro alkyl groups into phthalocyanines, as evidenced by the spectroscopic properties of 1,4,8,11,15,18,22,25-octa-fluoro 2,3,9,10,16,17,23,24-octa-perfluoro isopropyl zinc phthalocyanine, ZnF(64)Pc(-2) and its ring-reduced radical anion species, [ZnF(64)Pc(-3)](-), are reported. A combination of UV-visible absorption and magnetic circular dichroism (MCD) spectroscopy, ESI and MALDI-TOF mass spectrometry, cyclic and differential pulse voltammetry, and complete theoretical calculations using INDO/S and DFT techniques reveals that the substitution of all sixteen hydrogen atoms in protio ZnPc(-2) by eight F and eight i-C(3)F(7) groups red shifts the Q and pi --> pi transitions and narrows the HOMO-LUMO gap while simultaneously preventing ring photooxidation and stabilizing the radical anion. The [ZnF(64)Pc(-3)](-) species, which is in equilibrium in solution with the neutral complex when a reducing agent is present, is unusually stable. The above effects are attributed to the strong electron withdrawing properties of the peripheral substituents, which render ZnF(64)Pc extremely electron deficient. PMID- 12783562 TI - Peroxide electroreduction on bi-modified Au surfaces: vibrational spectroscopy and density functional calculations. AB - The mechanism of the electroreduction of peroxide on Bi-submonolayer-modified Au(111) surfaces is examined using surface-enhanced Raman scattering (SERS) measurements along with detailed density functional theory (DFT) calculations. The spectroscopy shows the presence of Bi-OH and Bi-O species at potentials just positive of that where peroxide is reduced. These species are not present in solutions absent either peroxide or Bi. DFT calculations show that peroxide is unstable relative to Bi-OH when Bi is present in the (2 x 2) configuration on Au(111) known from previous work to be catalytically active. The spacing between Bi adatoms is such that peroxide association with two Bi cannot occur without O-O bond cleavage. The full Bi monolayer is catalytically inactive and exhibits none of the Bi-OH or Bi-O signals seen for the active surface. The calculations show that as the Bi coverage becomes greater and the Bi adatom spacing becomes smaller, peroxide can adsorb on Bi without O-O bond rupture. These results indicate an important role for M-OH species in peroxide electroreduction. PMID- 12783563 TI - Alloyed semiconductor quantum dots: tuning the optical properties without changing the particle size. AB - Alloyed semiconductor quantum dots (cadmium selenium telluride) with both homogeneous and gradient internal structures have been prepared to achieve continuous tuning of the optical properties without changing the particle size. Our results demonstrate that composition and internal structure are two important parameters that can be used to tune the optical and electronic properties of multicomponent, alloyed quantum dots. A surprising finding is a nonlinear relationship between the composition and the absorption/emission energies, leading to new properties not obtainable from the parent binary systems. With red shifted light emission up to 850 nm and quantum yields up to 60%, this new class of alloyed quantum dots opens new possibilities in band gap engineering and in developing near-infrared fluorescent probes for in vivo molecular imaging and biomarker detection. PMID- 12783564 TI - Role of cluster size in catalysis: spectroscopic investigation of gamma-Al2O3 supported Ir4 and Ir6 during ethene hydrogenation. AB - gamma-Al(2)O(3)-supported Ir(4) and Ir(6) were prepared by decarbonylation of tetra- and hexanuclear iridium carbonyls, respectively, and compared as catalysts for ethene hydrogenation at atmospheric pressure and temperatures in the range 273-300 K. Rates of the reaction were determined along with extended X-ray absorption fine structure (EXAFS) and IR spectra characterizing the clusters in the working catalysts. EXAFS data show that the Ir(4) and Ir(6) cluster frames remained intact during catalysis. Di-sigma-bonded ethene and pi-bonded ethene on the clusters were identified by IR spectroscopy and found to compete as the principal reaction intermediates, with the former predominating at ethene partial pressures less than about 200 Torr and the latter at higher ethene partial pressures. Hydrogen on the clusters is inferred to form by dissociative adsorption of H(2); alternatively, it is provided by OH groups of the support. The rate of ethene hydrogenation on Ir(4) is typically several times greater than that on Ir(6). PMID- 12783565 TI - Diffusion anomaly as a function of molecular length of linear molecules: levitation effect. AB - Previous work on monatomic spherical sorbates has shown the existence of an anomalous peak in self-diffusivity (D) when plotted as a function of size of the diffusant. Molecular dynamics studies on linear molecules of different lengths l in zeolite NaY at 140 and 200 K are reported. It is seen that there is a peak in D as a function of l, suggesting that the levitation effect exists for linear molecules, the simplest member of polyatomics. This is confirmed by the lowering of the activation energy for the molecule whose length l exhibits highest D. Related quantities of interest such as the guest-host interaction energy and preexponential factor are discussed. PMID- 12783566 TI - Imaging structured water and bound polysaccharide on mica surface at ambient temperature. AB - The presence of a water layer on the surface of muscovite mica under ambient conditions is well established. The water molecules are well ordered and seem to be oriented, leading to an icelike monolayer (probably ferroelectric) in epitaxial relation with the mica surface. We have imaged and characterized the height and contact angle of ordered water layer(s) formed by wetting and de wetting processes on mica surfaces at different states of hydration by tapping mode atomic force microscopy. Implications that the presence of such an ordered water layer may have for imaging of biological samples are also discussed, with consideration of data for the polysaccharide hyaluronan. PMID- 12783567 TI - Theoretical study of the conversion of sulfonyl precursors into chains of poly(p phenylene vinylene). AB - The elimination and side reactions involved in the thermal conversion of sulfonyl precursor chains into poly(p-phenylene vinylene) (PPV) have been studied in detail, using Density Functional theory, along with the MPW1K functional. The performance of the MPW1K functional for describing radical dissociation and internal conversion reactions of sulfonyl precursors has been assessed against the results of benchmark CCSD(T) calculations. Enthalpies as well as entropies are calculated at different temperatures at the level of the rigid rotor-harmonic oscillator approximation. Entropy effects on internal elimination reactions are very limited. In sharp contrast, at the temperatures under which the conversion is usually performed (550 K), entropy contributions to the activation energies are found to be very significant and to strongly favor direct radical dissociations of the precursors. Further radical side reactions following an E(i) conversion through an alkyl substituent may also significantly contribute to the formation of sp(3) defects and/or cross-linked structures in the polymer-an advantageous feature for the making of materials with improved photoluminescence efficiencies. PMID- 12783568 TI - Modulation of the reduction potentials of fullerene derivatives. AB - The cyclic voltammetric (CV) study of a series of novel bisfulleropyrrolidines (3) and bisfulleropyrrolidinium ions (4) is reported. The eight possible stereoisomers of each series were systematically investigated under strictly aprotic conditions that allowed the observation of up to four and five subsequent reversible reductions in 3 and 4, respectively. Because of the stabilizing effect of positive charges, a significant enhancement of the electronegative properties was observed in 4. In fact, 4-trans-2 and 4-trans-1 result among the strongest reversible electron-accepting C(60) oligoadducts. Furthermore, the study evidenced that, in both 3 and 4, the CV pattern, and in particular the potential separation between the second and third reductions, changes significantly with the addition pattern. A sequential pi-electron model that simulates the effect of subsequent reductions of C(60) bis-adducts gives a good correlation (r > 0.96) with the cyclic voltammetry data when the molecules are divided in two sets dependent on the location of the addends in the same or in opposite hemispheres. PMID- 12783569 TI - Vancomycin resistance: occurrence, mechanisms and strategies to combat it. AB - Vancomycin has long been considered the antibiotic of last resort against serious and multi-drug-resistant infections caused by Gram-positive bacteria. However, vancomycin resistance has emerged, first in enterococci and, more recently, in Staphylococcus aureus. Here, the authors attempt to review the prevalence and the mechanisms of such resistance. Furthermore, they focus on strategies that have been developed or are under current investigation to overcome infections caused by vancomycin-resistant strains. Among these are glycopeptide derivatives with higher potency than vancomycin, small molecules that resensitise bacteria to the antibiotic and novel non-glycopeptide antibiotics. These agents are targeted to interfere with protein and/or peptidoglycan (PG) synthesis and integrity or with membrane permeability. Whilst most of these agents are still in clinical or preclinical development, some have entered the clinic and currently represent the only option for treating vancomycin-resistant enterococci (VRE). PMID- 12783570 TI - Minidefensins and other antimicrobial peptides: candidate anti-HIV microbicides. AB - Antimicrobial peptides have long been presumed to act as effector molecules of innate immunity. However, direct evidence that antimicrobial peptides have central roles in host defence has only recently become available. An overview of the types and characteristics of endogenous human antimicrobial peptides and proteins is presented, with particular emphasis on peptides that are active against HIV. These antiviral peptides are discussed in the context of utilising natural peptides for the design of effective topical microbicides for the treatment of sexually transmitted infections (STIs). Several antimicrobial peptides, termed minidefensins, are potently active against HIV, and bear structural similarity to their larger defensin cousins. Strategies to develop potent peptide antibiotics based on defensin and minidefensin templates are promising in the development of antiviral therapeutics and preventatives. PMID- 12783571 TI - Peripheral tachykinin receptors as potential therapeutic targets in visceral diseases. AB - More than 10 years of intensive preclinical investigation of selective tachykinin (TK) receptor antagonists has provided a rationale to the speculation that peripheral neurokinin (NK)-1, -2 and -3 receptors may be involved in the pathophysiology of various human diseases at the visceral level. In the airways, despite promising effects in animal models of asthma, pilot clinical trials with selective NK-1 or -2 receptor antagonists in asthmatics have been ambiguous, whereas the potential antitussive effects of NK-1, -2 or -3 antagonists have not yet been verified in humans. In the gastrointestinal (GI) tract, irritable bowel syndrome (IBS) and pancreatitis are appealing targets for peripherally-acting NK 1 and -2 antagonists, respectively. In the genito-urinary tract, NK-1 receptor antagonists could offer some protection against nephrotoxicity and cytotoxicity induced by chemotherapeutic agents, whereas NK-2 receptor antagonists appear to be promising new agents for the treatment of neurogenic bladder hyperreflexia. Finally, there is preclinical evidence for hypothesising an effect of NK-3 receptor antagonists on the cardiovascular disturbance that characterises pre eclampsia. Other more speculative applications are also mentioned. PMID- 12783572 TI - The potential of antisense as a CNS therapeutic. AB - Antisense offers a precise and specific means of knocking down expression of a target gene, and is a major focus of research in neuroscience and other areas. It has application as a tool in gene function and target validation studies and is emerging as a therapeutic technology in its own right. It has become increasingly obvious, however, that there are a number of hurdles to overcome before antisense can be used effectively in the CNS, most notably finding suitable nucleic acid chemistries and an effective delivery vehicle to transport antisense oligonucleotides (AS-ODNs) across the blood-brain barrier (BBB) to their site of action. Despite these problems, a number of potential applications of AS-ODNs in CNS therapeutics have been validated in vitro and, in some cases, in vivo. Here the authors outline available nucleic acid chemistries and review progress in the development of non-invasive delivery vehicles that may be applicable to CNS therapeutics. Further to this, they discuss a number of experimental applications of AS-ODNs to CNS research and speculate on the development of antisense techniques to treat CNS disease. PMID- 12783573 TI - New therapeutic target in primary headaches - blocking theCGRP receptor. AB - The primary headaches are among the most prevalent neurological disorders, afflicting up to 16% of the adult population. The associated pain originates from intracranial blood vessels that are innervated by sensory nerves storing several neurotransmitters. In primary headaches, there is a clear association between the headache and the release of calcitonin gene-related peptide (CGRP), but not other neuronal messengers. The specific purpose of this review is to describe CGRP in the human cranial circulation and to elucidate a possible role for a specific antagonist in the treatment of primary headaches. Acute treatment with administration of a 5-HT(1B/1D) agonist (triptan) results in alleviation of the headache and normalisation of the CGRP level. The mechanism of action of triptans involves vasoconstriction of intracranial vessels and a presynaptic inhibitory effect of sensory nerves. The central role of CGRP in migraine and cluster headache pathophysiology has led to the search for small-molecule CGRP antagonists, which are predicted to have fewer cardiovascular side effects in comparison to the triptans. The initial pharmacological profile of such a group of compounds has recently been disclosed. These compounds have high selectivity for human CGRP receptors and are reportedly efficacious in the relief of acute attacks of migraine. PMID- 12783574 TI - Matrix metalloproteinase inhibitors for cancer therapy:the current situation and future prospects. AB - Inhibition of matrix metalloproteinases (MMPs), a family of proteolytic enzymes linked to many aspects of cancer progression, has been explored as a therapeutic goal for almost two decades. Thus far, all tested MMP inhibitors (MMPIs) have failed to reach primary end points in Phase III clinical trials, although secondary analyses suggest benefits in particular patient groups. The clinical development of these agents has been hampered by problems related to determination of effective dosages and side effects that necessitate dose lowering or drug holidays. Imaging technologies offer hope as a means to measure enzyme activity and hence effective enzyme inhibition in vivo. Meanwhile, recent results from genetic studies of both mice and man have given some clues to possible causes of musculoskeletal side effects. Future progress in the therapeutic use of MMPIs is dependent on the ability to selectively target cancer associated MMPs at the correct stage in tumour progression and the development of surrogate markers of in vivo efficacy. PMID- 12783575 TI - The coagulation system as a target for experimental therapy of human gliomas. AB - The purpose of this paper is to review the rationale for the development of coagulation-reactive drugs for the experimental therapy of gliomas. Numerous reactants familiar to students of blood coagulation have been shown to contribute to neoplastic proliferation, invasion and metastasis. Recently, considerable progress has been made in demonstrating the ability of drugs capable of inhibiting these reactants to alter cancer progression. Biological features of gliomas within the realm of blood coagulation suggest that clinical trials of such drugs warrant consideration. This approach offers the prospect of a novel treatment for this devastating tumour type that does not share the toxicities of conventional cancer therapies. PMID- 12783576 TI - Antiangiogenic therapy through copper chelation. AB - As new compounds are being evaluated for use in clinical trials involving antiangiogenic therapies, two important factors must be considered. Independent of clinical efficacy, the potential drug must be cost-effective and have reasonable ease of production. The compound endostatin (Entremed, Inc.) has recently completed two Phase I trials with minimal toxicity to the patients treated [1,2]. However, due to the difficulty and expense of producing large quantities of a recombinant protein, Entremed Inc. has experienced financial difficulties [3]. As this company's fate indicates, a drug must not only be clinically effective, but must also possess reasonable production economics. Another interesting component of compound development is selectivity. Highly selective antiangiogenic compounds such as the tyrosine kinase inhibitor SU-5416 are being replaced by less selective compounds such as SU-6668, which acts on a broader spectrum of tyrosine kinase receptors [4]. This move towards using less selective antiangiogenic compounds is based on preclinical models that demonstrate both better clinical efficacy when using less specific molecules and low response rates from the more selective compounds. With the aim of further examining broadly-acting antiangiogenic agents, the authors are currently evaluating new classes of agents that preferentially bind copper and inhibit angiogenesis. Copper has been known to be a significant target for antiangiogenic therapy for a number of years [5]. Recently, through the use of molecular techniques, the target enzymes that utilise copper as a cofactor are being elucidated. This review will describe the historical use of anticopper therapy for the treatment of Wilson's disease and evaluate some of the new anticopper compounds currently under consideration for use in antiangiogenic therapy. PMID- 12783577 TI - TNF-receptor-associated factors as targets for drug development. AB - TNF-receptor-associated factors (TRAFs) are the bottleneck of the TNF-receptor (TNF-R) family signal transduction. They integrate the signalling from many members of the TNF-R family and initiate intracellular signalling cascades aimed at the activation of NF-kappaB and c-jun, the reprogramming of gene expression and the control of cell death. Deregulation of these pathways is the cause of several autoimmune and inflammatory diseases. The specificity and interaction of the members of the TRAF family with the TNF-R entails the recognition of just a 4 - 6 amino acid motif in the cytosolic region of the receptor, suitable as an attractive target for drug discovery. This review summarises the current knowledge on TRAFs and discusses the pros and cons of their application as targets for drug discovery. PMID- 12783578 TI - Secreted phospholipase A2 enzymes as therapeutic targets. AB - Homology cloning through in silico database search analysis has led to the definition of ten structurally-related mammalian secreted phospholipase A(2) (sPLA(2)) enzyme forms at present, each expressed in a species-, genotype- and cell-type-specific manner and with different enzymatic properties. These studies have shown that models based on the premise that there is only one PLA(2) drug target are now inadequate. Type IIA sPLA(2) remains the most advanced clinical target, with rationally designed inhibitors in Phase II clinical trials. However, progress in our understanding of the functional role of the ten secreted enzymes in phospholipid (PL) metabolism and in eicosanoid-mediated disorders, together with their emerging activity-independent and receptor-mediated functions, is likely to significantly impact on current and future drug development efforts. PMID- 12783579 TI - Isoform-specific regulation of adenylyl cyclase: a potential target in future pharmacotherapy. AB - Adenylyl cyclase (AC) is a target enzyme of multiple G-protein-coupled receptors (GPCRs). In the past decade, the cloning, structure and biochemical properties of nine AC isoforms were reported, and each isoform of AC shows distinct patterns of tissue distribution and biochemical/pharmacological properties. In addition to the conventional regulators of this enzyme, such as calmodulin (CaM) or PKC, novel regulators, for example, caveolin, have been identified. Most importantly, these regulators work on AC in an isoform dependent manner. Recent studies have demonstrated that certain classic AC inhibitors, i.e., P-site inhibitors, show an isoform-dependent inhibition of AC. The side chain modifications of forskolin, a diterpene extract from Coleus forskolii, markedly enhance its isoform selectivity. When taken together, these findings suggest that it is feasible to develop new pharmacotherapeutic agents that target AC isoforms to regulate various neurohormonal signals in a highly tissue-/organ-specific manner. PMID- 12783580 TI - The ATPases: a new family for a family-based drug design approach. AB - The rapid discovery of new drugs is greatly facilitated when a family of related proteins is targeted with a similar approach in chemistry. Few protein families have so far been investigated using this kind of 'family-based' approach. Therefore, to increase the size of our Pharmacopeia and to cure human diseases more efficiently, new druggable protein families must be identified. It is shown in this review that ATPases are very good candidates for a family-based approach. The human proteome contains many ATPases, which are involved in several diseases. All the ATPases contain a nucleotide-binding site, and it is therefore possible to target all of them with a single strategy in chemistry; the design of competitive ATP inhibitors. Moreover, because a similar approach has been conducted with the protein kinases, the compound libraries and the knowledge developed in the kinase field can be directly applied to the ATPases. PMID- 12783581 TI - Neoadjuvant chemotherapy and chemoradiotherapy for non-small cell lung cancer: current status and future prospects. AB - Of the patients with non-small cell lung cancer, 60% present with localised or locally advanced disease. Although they may be considered potentially curable, the vast majority will die, usually from systemic disease. So far, adjuvant (postoperative) therapy has failed to demonstrate benefit. In contrast, chemotherapy has demonstrated clear advantages when administered prior to, or concurrently with radiotherapy in Stage III disease or prior to surgery in Stage III disease. Chemotherapy administered prior to surgery, termed neoadjuvant therapy, in Stage I and II disease has been demonstrated to be feasible. Several trials employing currently available agents have yielded promising results. Whether these regimens will result in an unequivocal benefit is the subject of several ongoing studies in the US and Europe. Current research is focusing on the role of newer drugs including novel antitubulin agents, growth factor receptor antagonists, eicosanoid modulators and various other agents. PMID- 12783582 TI - Neoadjuvant chemotherapy for bladder cancer: current status. AB - Muscle-invasive bladder cancer is typically an aggressive solid tumour with the propensity for early systemic dissemination. Although curative-intent local therapy with either radical cystectomy or radiotherapy remains the gold standard intervention, the high rate of systemic failure has prompted investigators to evaluate various strategies in an attempt to improve survival, including the early administration of systemic chemotherapy. Recently completed, Phase III studies of neoadjuvant chemotherapy trials demonstrated a modest, but real survival advantage for a small subset of patients. Other strategies include, attempts to both preserve the bladder using combinations of limited surgical resection, systemic chemotherapy and radiotherapy. This review focuses on the potential of neoadjuvantly-administered therapies to impact the management of muscle invasive bladder cancer. PMID- 12783583 TI - Neoadjuvant chemotherapy for cervical cancer. AB - None of the current surgical or radiation treatment strategies for cervical cancer satisfactorily leads to a high disease-free survival and a low risk for treatment-related complications in patients with bulky or locally-advanced disease. Neoadjuvant chemotherapy (NACT) prior to surgery or radiation therapy has been studied as a means to reduce tumour bulk and thereby rendering subsequent therapy more effective. Impressive clinical response rates to cisplatin-based NACT have been achieved with acceptable toxicity and survival. Of the patients treated, approximately 20% will achieve a complete clinical response and many of these patients will prove to have a complete pathological response. Although there are too few randomised controlled studies to determine the effectiveness of NACT approaches, relative to standard treatments. PMID- 12783584 TI - Pharmacotherapy of gastrointestinal stromal tumours. AB - Gastrointestinal stromal tumours (GIST) are a relatively rare, but well characterised clinical entity. This tumour is defined by a predominantly spindle cell morphology and its pathobiology by the presence of Kit (receptor tyrosine kinase). The majority of GIST have a gain of function mutation in an exon hot spot that leads to constitutive activation of Kit, promoting proliferation and anti-apoptotic signalling. Imatinib mesylate (Gleevec), Glivec, Novartis) is a specific inhibitor of Kit kinase activation and in Phase II clinical trials, it has proven to be remarkably effective in heavily pre-treated patients with advanced GIST. Molecular determinants of response and resistance are the subject of ongoing investigations. Additionally, clinical trials are underway to explore the use of imatinib mesylate in the adjuvant setting. These initial evaluations with imatinib mesylate provide proof of concept for hypothesis driven, rational drug design of selective signal transduction inhibitors in the management of solid tumour malignancies. PMID- 12783585 TI - Pharmacotherapy of hormone refractory prostate cancer: new developments and challenges. AB - Hormone refractory prostate cancer (HRPC) remains a challenge in the management of prostate cancer patients. With the widespread use of PSA (prostate specific antigen), recurrent disease after local treatment for localised prostate cancer is usually diagnosed long before evidence of metastatic disease. In many cases, hormonal manipulations are started at the time of biochemical relapse and therefore, patients become 'hormone refractory' earlier in the course of their disease, frequently with a good performance status, often with no evidence of metastatic disease, and they still face a considerably long life expectancy. Despite these changes, the need for more options in the treatment of HRPC is obvious. The pharmacological treatments that are in use and those that are under investigation for this group of patients will be discussed and include: cytotoxic agents including the microtubule inhibitors, alone and in combination with other conventional or experimental therapies such as calcitriol or thalidomide; treatment with epothilone analogues; endothelin receptor antagonists; palliative therapy with bisphosphonates, bone-targeted radiopharmaceuticals and other developing treatments such as vaccines, gene therapies and monoclonal antibodies. PMID- 12783586 TI - Toxicity of platinum compounds. AB - Since the introduction of platinum-based combination chemotherapy, particularly cisplatin, the outcome of the treatment of many solid tumours has changed. The leading platinum compounds in cancer chemotherapy are cisplatin, carboplatin and oxaliplatin. They share some structural similarities; however, there are marked differences between them in therapeutic use, pharmacokinetics and adverse effects profiles [1-4]. Compared to cisplatin, carboplatin has inferior efficacy in germ cell tumour, head and neck cancer and bladder and oesophageal carcinoma, whereas both drugs seem to have comparable efficacy in advanced non-small cell and small cell lung cancer as well as ovarian cancer [5-7]. Oxaliplatin belongs to the group of diaminocyclohexane platinum compounds. It is the first platinum-based drug that has marked efficacy in colorectal cancer when given in combination with 5-fluorouracil and folinic acid [8,9]. Other platinum compounds such as oral JM216, ZD0473, BBR3464 and SPI-77, which is a pegylated liposomal formulation of cisplatin, are still under investigation [10-13], whereas nedaplatin has been approved in Japan for the treatment of non-small cell lung cancer and other solid tumours. This review focuses on cisplatin, carboplatin and oxaliplatin. PMID- 12783588 TI - Androgen replacement therapy in the hypogonadal ageing man. AB - In men, gonadal function is affected in a slow, progressive way as part of the normal ageing process. Recently, however, significant interest has developed on the importance of this condition, which is variously known as male climacteric, andropause or, more appropriately, androgen decline in the ageing male (ADAM). The term andropause is biologically wrong and clinically inappropriate but, it adequately conveys the concept of emotional and physical changes that, although related to ageing in general, are also associated with significant hormonal alterations. The inappropriateness of the term is based on the fact that in women, the reproductive cycle invariably ends with ovarian failure. In men, this process is not universal and when it occurs it is normally subtle in its clinical manifestations. This has led to a tendency to ignore the syndrome as an unavoidable and untreatable result of the ageing process. For the sake of simplicity and directness, this review will use the terms ADAM and andropause to denote the global hormonal alterations associated with ageing. PMID- 12783587 TI - Pharmacotherapy of carpal tunnel syndrome. AB - Carpal tunnel syndrome (CTS) is the most common peripheral nerve entrapment syndrome. CTS is a compression neuropathy caused by elevated pressure in the carpal tunnel. CTS has the potential to substantially limit performance of activities of daily living for some individuals. The goal of therapy for CTS is to improve symptoms and reduce signs of the disease, as well as prevent progression and loss of hand function. There are several treatment alternatives to relieve the pressure on the median nerve, both surgical and conservative. The most common measures employed in the initial treatment of CTS are NSAIDs, local and systemic corticosteroids, diuretics and pyridoxine. However, CTS treatment usually includes a combination of pharmacotherapy with other strategies such as splinting and activity modification. Injections of corticosteroids into the carpal tunnel are often employed for cases not responding to conservative treatment. Surgery is superior to conservative therapies for most persistently symptomatic patients. The aim of this paper is to review the pharmacological agents used for relieving the symptoms of CTS. PMID- 12783589 TI - Pharmacotherapy of heparin- induced thrombocytopenia. AB - Heparin-induced thrombocytopenia (HIT) is a life-and-limb threatening condition that is associated with the development of antibodies that activate platelets and the coagulation system in the presence of unfractionated heparin or low molecular weight heparin. The binding of antibody to heparin-PF-4 complexes can activate platelets, leading to an acute, often catastrophic, thrombotic diathesis. The most common laboratory finding is the development of thrombocytopenia 5 or more days after beginning heparin treatment, which occur in up to 1 - 5% of patients exposed to heparin, depending on type of heparin and indication for anticoagulation. The onset of thrombocytopenia can be immediate or delayed for several weeks after the exposure to heparin. Approximately 50 - 60% of patients who develop HIT manifest acute venous or arterial thrombosis and a significant percentage of these patients die or develop vascular gangrene of a limb that requires amputation. Given the severe sequelae associated with HIT, recognition and immediate medical management is essential. Treatment of a patient with HIT is complex, as there are several different anticoagulants now available which have been shown to be useful. Optimal management depends on each patient's individual clinical manifestations, as well as the need for ongoing anticoagulation therapy. No single agent or treatment approach can be considered to be 'standard practice' as very few clinical trials have been completed, compare different treatment options. The use of warfarin alone in a patient with HIT, must be avoided in order to avoid the possibility of further activating coagulation, which may hasten the development of venous limb gangrene. There are several different tests available that detect HIT antibodies and each has different sensitivity and specificity for HIT. In this review we discuss the epidemiology and natural history of HIT, risk factors associated with the development of HIT and the clinical and laboratory tests that aid in the diagnosis and treatment. Special emphasis is given to addressing the management of HIT in special populations, particularly patients with renal or liver disease, acute coronary syndromes, pregnancy, paediatrics and patients who require cardiopulmonary bypass surgery. PMID- 12783590 TI - Phosphate binder usage in kidney failure patients. AB - Phosphorus binders are used in patients with kidney failure because of the incomplete removal of phosphorus with dialysis and the inability to exclude phosphorus from the diet. Aluminium was the initial phosphorus binder used, but was replaced by calcium-containing binders because of the development of aluminium toxicity. Calcium-based binders have been the mainstay of therapy for many years, but recent investigations have pointed to increased rates of vascular calcification in patients taking calcium-containing binders. For this reason, alternative agents have been developed. Sevelamer (Renagel), GelTex Pharmaceuticals Inc.) is a polymer which has been found to effectively bind phosphorus. It has resulted in a decreased rate of vascular calcification compared to calcium-containing binders. Other agents under development include lanthanum carbonate and iron-complex preparations. Further research will likely concentrate on identifying binders that bind phosphate more efficiently, have minimal gastrointestinal side effects and provide other benefits to dialysis patients. PMID- 12783591 TI - Prescription drugs and pregnancy. AB - Prescribing drugs in pregnancy is an unusual risk-benefit situation. Drugs that may be of benefit or even life-saving to the mother can deform or kill the fetus. However, the risk to the fetus should not be exaggerated. There are only approximately 20 drugs or groups of drugs which are known to cause birth defects in humans. For one of these drugs to cause birth defects, a number of criteria must be fulfilled. The drug exposure must take place at a critical stage of pregnancy and the dose must be high enough to cause a threshold of exposure for an appropriate duration of time. For most of the known human teratogens, > 90% of pregnancies exposed during the first trimester result in normal offspring. Although only a few drugs are known to cause birth defects in humans, uncertainty about the safety of the majority may lead to underprescribing for pregnant women and women of childbearing age. Epidemiological studies of pregnancy outcome after specific drug exposures are often superficially reassuring, but most are severely limited in their power to detect adverse outcomes. Safety in animal studies may also be reassuring but species differences demand caution in this interpretation. Concerns about prescription drugs in the first trimester, when they can cause birth defects, are mostly quite different to concerns about use in the second and third trimesters. As the fetal organ systems mature, the fetus can be affected by the pharmacological activity of the drug in the same way as the mother. Many drugs have pharmacological effects on the fetus in the second and third trimesters but in most cases, they are well recognised and can be managed or avoided. The material presented in this paper is mostly concerned with the 'risks' associated with drugs in pregnancy. No attempt has been made to quantitate the possible benefits to the mother or fetus. Communicating the risk benefit situation to the patient is always a challenge for physicians with limited time and sometimes limited knowledge. Fear of litigation is an unfortunate and an unwanted parameter in the assessment. Better knowledge of the parameters that determine teratogenicity may allow physicians to feel more confident in assessing the risks and benefits associated with prescribing in pregnancy. PMID- 12783592 TI - Imatinib mesylate in the treatment of chronic myeloid leukaemia. AB - Imatinib mesylate (Gleevec, Glivec), formerly STI571; Novartis Pharmaceuticals) is an inhibitor of the Bcr-abl tyrosine kinase that is central to the pathogenesis of chronic myeloid leukaemia (CML). The remarkable results of imatinib mesylate in clinical trials have rapidly and profoundly changed the management of patients with CML. This article will review the development of this molecularly targeted agent. The clinical trials with imatinib mesylate will be summarised along with the pharmacology of this agent. Despite the impressive responses seen in chronic-phase patients, numerous questions remain. For example, how durable will responses to imatinib mesylate be and is it necessary or possible to improve upon these results? Ongoing efforts to address these issues will be discussed. PMID- 12783593 TI - Dofetilide: a new drug to control cardiac arrhythmia. AB - Atrial fibrillation (AF) is the most common cardiac arrhythmia. Mortality, and especially morbidity caused by AF, are major and growing health problems in the western world. AF is strongly associated with arterial hypertension, congestive heart failure, valvular heart disease, ischaemic heart disease, and with prevalence increasing with age. A variety of drugs have been used to terminate or prevent AF but, as many antiarrhythmic agents have the potential life-threatening pro-arrhythmia, safety problems remain. Dofetilide (Tikosyn, Pfizer), a new Vaughan Williams class III antiarrhythmic agent, has been developed and approved for the treatment of AF. In contrast to most antiarrhythmic agents, the development programme included two safety studies in high-risk patients. Dofetilide is effective and safe when an elaborate procedure for dosing is implemented. Along with amiodarone and betablockers, dofetilide is the only antiarrhythmic drug, which is recommended by guidelines for the treatment of AF in a wide range of patients. PMID- 12783594 TI - Leflunomide for the treatment of rheumatoid arthritis. AB - Leflunomide (Arava), Aventis Pharmaceuticals) is an immunomodulating drug that interferes with the metabolism of pyrimidine by inhibiting dihydro-orotate dehydrogenase (DHO-DH) in mitochondria, thereby blocking T- and B cell proliferation. Antibody production is also affected by DHO-DH blockade. Other immunomodulatory effects of leflunomide have also been reported. Symptomatic and structural effects of leflunomide in active rheumatoid arthritis have been strictly evaluated by double-blind, randomised, placebo-controlled studies that were aimed at validating its use in rheumatoid arthritis. Further trials are now required to confirm efficacy and safety of this drug in combination with other agents. PMID- 12783595 TI - Is natalizumab a breakthrough in the treatment of multiple sclerosis? AB - In patients with either relapsing-remitting or secondary-progressive multiple sclerosis, there were fewer new lesions/patient with natalizumab (0.7 and 1.1 with natalizumab 3 and 6 mg/kg every 28 days, respectively) than in the placebo group (9.6 new lesions/patient) over 6 months. There were also fewer relapses in the natalizumab groups than the placebo group. However, there were no changes in the Expanded Disability Status Scale scores in any of the groups. Natalizumab was well-tolerated. Thus, the initial results with natalizumab treatment over 6 months in multiple sclerosis are encouraging. PMID- 12783596 TI - Novel pharmacological approaches in the treatment of breast cancer. AB - Breast cancer is the most common malignancy among women. Novel pharmacological agents, including hormonal, cytotoxic and biological therapies, are currently being developed and tested in clinical trials and may offer patients more treatment options and an improved chance of long-term survival. Signal transduction inhibitors that block endocrine or growth factor pathways have demonstrated exciting antitumour effects in clinical trials. In addition to new chemotherapeutic drugs, numerous biological agents including growth factor receptor-directed monoclonal antibodies and tyrosine kinase inhibitors that target specific molecular lesions are being examined as potential breast cancer treatments. PMID- 12783597 TI - The therapeutic potential of novel antiangiogenic therapies. AB - Promising new antiangiogenic strategies are emerging for the treatment of cancer. Numerous candidate drugs that target vascular endothelial growth factor, vascular endothelial growth factor receptors, integrins, matrix metalloproteinases and other blood vessel targets are being developed and tested in clinical trials. This review highlights the numerous drugs in clinical trials and expands on potential new approaches to inhibiting angiogenesis. These approaches include gene therapy, vaccine strategies and antiangiogenic radioligands. New insight has been gained from completed Phase III trials with antiangiogenic drugs and some of the major obstacles include design of trials, dosing, toxicities and resistance. This review will discuss these barriers and methods by which they can be overcome. PMID- 12783598 TI - Angiopoietins in angiogenesis and beyond. AB - The angiopoietin (Ang) family of growth factors includes four members, all of which bind to the endothelial receptor tyrosine kinase Tie2. Two of the Angs, Ang 1 and Ang-4, activate the Tie2 receptor, whereas Ang-2 and Ang-3 inhibit Ang-1 induced Tie2 phosphorylation. While genetic models have underscored the importance of Angs in the developing cardiovascular system, other studies have demonstrated that Ang-1 promotes endothelial cell survival, sprouting and tube formation. More recently, a new aspect of the biology of this class of growth factors has emerged, namely the ability of Ang-1 to reduce inflammation. This review presents an outline of Angs and their receptors, examining their structure, expression, signalling, regulation and biological significance and comments on the role and potential usefulness of Angs in medicine. PMID- 12783599 TI - Farnesyl transferase inhibitors in clinical development. AB - Farnesyl transferase (FT) inhibitors block the main post-translational modification of the Ras protein, thus interfering with its localisation to the inner surface of the plasma membrane and subsequent activation of downstream effectors. Although initially developed as a strategy to target Ras in cancer, FT inhibitors have subsequently been acknowledged as acting by additional and more complex mechanisms that may extend beyond Ras, involving RhoB, centromere-binding proteins and probably other farnesylated proteins. SCH66336 (lonafarnib, Sarasar( trade mark ); Schering-Plough), a tricyclic orally active FT inhibitor, was the first of these compounds to undergo clinical development. Gastrointestinal tract toxicities and fatigue have qualified as dose-limiting toxicities in all Phase I/II studies. Evidence of clinical activity has been reported. Lonafarnib combination studies with both gemcitabine and paclitaxel have been carried out. No unexpected toxicities were observed in these Phase I studies, while encouraging clinical activity was observed mainly in pancreatic cancer and non small cell lung cancer. Further combination studies are ongoing. R115777 (Zarnestra( trade mark ); Janssen Pharmaceutica) is another orally active FT competitive inhibitor in clinical development. Single-agent Phase I/II studies have shown that myelotoxicity and neurotoxicity are dose-limiting toxicities; intermittent schedule is probably better tolerated; antitumour activity is observed particularly in breast cancer and haematological malignancies. A number of combination studies with R115777 have been carried out. A recently completed, large Phase III trial comparing gemcitabine plus R115777 versus gemcitabine plus placebo in advanced pancreatic cancer has failed to demonstrate any survival benefit in the R115777 arm. BMS-214662 is the third FT inhibitor in clinical development. It has the main advantage of being cytotoxic in nature, rather than cytostatic, and potent in vivo antitumour activity has been reported. A major drawback for BMS-214662 is its severe gastrointestinal and liver toxicities, which prevent the achievement of adequate systemic exposures following the oral route. Alternative ways of interference with the ras oncogene pathway, in particular inhibition of ras downstream effectors, are discussed and early clinical data are presented. PMID- 12783600 TI - CDK inhibitors in clinical development for the treatment of cancer. AB - Cyclin-dependent protein kinases (CDKs) are key regulators of the cell division cycle, whose various checkpoints proliferating cells must traverse. Since CDK deregulation, either through direct or indirect means, is found in most cancer cells, pharmacological CDK inhibition has become an attractive strategy towards mechanism-based and non-genotoxic therapies in oncology. Over the last decade, discovery and lead optimisation efforts have provided a wealth of potential drug candidate molecules capable of inhibiting CDKs, blocking cell-cycle progression, modulating transcription and inducing apoptosis selectively in cancer cells. However, only few such agents have as yet reached clinical evaluation. Here, the preclinical and clinical results obtained so far with flavopiridol (L868275, HMR1275; Aventis), 7-hydroxystaurosporine (UCN-01, KW-2401; Kyowa Hakko Kogyo) and roscovitine (R-roscovitine, CYC202; Cyclacel) are summarised. Furthermore, the potential for monotherapy and applications in combination with existing drugs are discussed. PMID- 12783601 TI - Recent progress in tumour vaccine development. AB - Immunotherapy offers an exciting opportunity to treat human cancer. Analysis of tumour-associated antigens is progressing. Assisted by animal models, such knowledge can be used to design tumour vaccines. By including adjuvants to increase immunogenicity, several tumours previously thought to be non-immunogenic are now considered targets for tumour vaccines. Newly acquired knowledge regarding dendritic cell physiology is incorporated in newly designed vaccines that are currently in Phase I and II trials. Such assessment provides the overall conclusion that tumour vaccines are safe and deserve a more prominent place in the sequel of treatments for human cancer. PMID- 12783602 TI - Recent developments in the pharmacological treatment of advanced pancreatic cancer. AB - Metastatic pancreatic cancer is one of the leading causes of cancer-related death in North America and Europe. The high mortality rate associated with pancreatic cancer is related to the fact that the vast majority of patients develop incurable, metastatic disease. Such patients have, in the past, had few treatment options. In recent years, however, the systemic administration of gemcitabine has been accepted as a standard first-line treatment for patients with advanced pancreatic cancer. While treatment with gemcitabine has been shown to result in both clinical benefit and in prolongation of survival, objective tumour responses following therapy with gemcitabine are relatively uncommon and median survival times remain short. Current efforts have, therefore, focused on evaluating chemotherapy regimens in which gemcitabine is combined with a second cytotoxic agent. Several such combinations appear to be associated with higher objective response rates than single-agent gemcitabine and have been well-tolerated in early clinical trials. Ongoing, prospectively randomised clinical trials will help better define the efficacy of these new combinations and will determine if they result in a significant benefit when compared to gemcitabine monotherapy. A number of novel chemotherapeutic and biological agents also appear promising and are likely to play a future role in the treatment of patients with advanced pancreatic cancer. PMID- 12783603 TI - Current and potential pharmacological treatments for obsessive-compulsive disorder. AB - In the past 20 years, several effective treatments have been developed for obsessive-compulsive disorder. Despite this, it is a disorder that is often inadequately treated by available therapies. Recent advances in neuroimaging and neurophysiology have provided some clues to possible treatments that may be more effective in treating this disorder. Serotonin agents remain the cornerstone of pharmacological treatment for obsessive-compulsive disorder. The development of agents that target specific serotonin receptor subtypes may improve the effectiveness of drug treatment. Other possible future treatment approaches are also discussed. PMID- 12783604 TI - Current and novel therapies for Lyme disease. AB - From tick bite to treatment, Lyme disease is a serious concern for those who reside in parts of the world where this infectious disorder is endemic. It is a multi-system and sometimes multi-stage illness caused by a unique spirochetal bacterium, Borrelia burgdorferi, which reaches its human victim after being transmitted by a bite from Ixodid (deer) ticks that are found mostly in certain parts of North America and Europe. Management of Lyme disease patients is usually not problematic, especially in endemic areas, where rapid diagnosis and response to the recommended treatment regimen most often leads to a favourable outcome. Initially this review article describes briefly some of the key historical, epidemiological, microbiological and diagnostic aspects of Lyme disease. With this foundation, there follows a more in-depth coverage on the proper management of asymptomatic tick bite victims and of those suffering from authentic Lyme disease. Much of the key information related to proper therapy is based on results that have accrued from rigorously conducted clinical studies. The final part of this review then concentrates on some of the more controversial issues involving diagnosis and treatment, the availability and possible usefulness of other non-antibiotic treatment options and the prospects for more efficient antibiotic treatment regimens as well as development of a newer class of effective antibiotics, based on promising preclinical data. PMID- 12783605 TI - Dehydroepiandrosterone, a sex steroid metabolite in development for systemic lupus erythematosus. AB - Deficiency of the weak androgen dehydroepiandrosterone (DHEA) and its sulfoconjugated metabolite DHEA-S has been associated with a number of serious illnesses, including lupus, diabetes, Alzheimer's disease and some cancers. Accordingly, supplementation with DHEA has been proposed for a variety of illnesses. Observational clinical studies and in vitro experiments have suggested that DHEA treatment might have a significant impact on immunological function, bone density, cognition, atherosclerotic disease, some malignancies, insulin resistance and obesity. Endogenous circulating DHEA levels, however, may vary widely by gender, age and ethnicity and can be affected by acute changes in corticosteroid production, alcohol intake, smoking, body mass index, medications and thyroid function [1-3]. Clearly, these variables complicate the interpretation of clinical data. DHEA also gives rise to a number of as yet poorly characterised metabolites, further confusing the assessment of its net effects when considered as treatment in heterogenous populations. Given the complexity of potential effects of DHEA and its metabolites, coupled to the diversity of clinical conditions that they might, at least in theory, affect, it is not surprising that clinical confirmation of efficacy in several clinical contexts has been inconsistent and controversial, hampering drug development in what might potentially be an important and widespread market. The current review will consider recent work suggesting efficacy of DHEA (GL-701, prasterone, Prestara( trade mark ) [US], Anastar( trade mark ) [Europe]; Genelabs) in systemic lupus erythematosus. PMID- 12783606 TI - Evolving role of bivalirudin in percutaneous coronary interventions; impact of the REPLACE-2 study. AB - Percutaneous coronary interventions are being used with increasing frequency nowadays for the treatment of patients with coronary heart disease. Acute thrombotic complications remain one of the major limitations of these procedures for which unfractionated heparin has been the standard foundation anticoagulant. It has, however, many limitations and disadvantages that necessitated research and development of alternative anticoagulant therapies. The direct thrombin inhibitor bivalirudin has been approved as a substitute for heparin in patients undergoing angioplasty for unstable angina based on data in higher-risk patients where bivalirudin resulted in lower rates of ischaemic and bleeding complications compared to heparin. This evidence was collected prior to the widespread use of platelet glycoprotein (GP) IIb/IIIa receptor blockers, thienopyridines and intracoronary stents, considered standard practice for such patients today. The REPLACE-2 study was carried out to establish whether, in the current era, bivalirudin used with provisional GP IIb/IIIa blockade if necessary during the procedure could provide equivalent protection from ischaemic events compared with the gold standard of heparin plus routine GP IIb/IIIa blockade. With advantages with regards to bleeding, ease of use and reduced cost, bivalirudin use with provisional GP IIb/IIIa inhibitors in low-to-moderate risk percutaneous coronary interventions allows GP IIb/IIIa receptor antagonists to be used in a selective fashion, rather than in all patients. In this article, background rationale for bivalirudin use in this setting is reviewed as well as past research in this area. Additionally, the implications of the REPLACE-2 study in establishing where bivalirudin fits into our current interventional cardiology practice and future directions are presented. PMID- 12783608 TI - CNIO cancer conference: targeted search for anticancer drugs. AB - The topics discussed at the conference covered many aspects of cancer research, from the genetic search for new targets, target validation and drug discovery, all the way to preclinical and clinical development of oncology drugs. Here the presentations on new metabolic, angiogenic, cell cycle and other molecular targets, as well as recent developments with experimental drugs with action on some of these targets, are summarised. Particular emphasis is placed on the emerging realisation that changes in the metabolic phenotype lie at the heart of cellular transformation. New insights into the biological links between cancer cell metabolism and the balance between survival and death signalling are likely to lead to the identification of a new category of anticancer targets. PMID- 12783607 TI - Does the melanin-concentrating hormone antagonist SNAP-7941 deserve 3As? AB - Melanin-concentrating hormone (MCH) is orexigenic (stimulates food intake). Two receptors for MCH have been identified in humans, MCH1-R and MCH2-R. SNAP-7941 is a small molecule MCH1-R antagonist. SNAP-7941 inhibits MCH-induced food intake in rats. SNAP-7941 alone reduced weight gain in young growing rats and in mature rats fed a high-fat diet. Preliminary testing with SNAP7941 in animal models of depression and anxiety shows it has antidepressant and anxiolytic effects. SNAP7941 should undergo further development as an anorectic, antidepressant and anxiolytic. PMID- 12783609 TI - Liver stem cells: prospects for treatment of inherited and acquired liver diseases. AB - It is now understood that there are three cell compartments which physiologically contribute to vertebrate liver parenchymal maintenance and regeneration after injury: mature liver cells (hepatocytes, cholangiocytes), intraorgan stem/progenitor cells (cells of the proximal biliary tree, periductal cells) and extraorgan stem cells (from the circulation and the bone marrow). All of these cell populations, as well as other, non-physiologic stem cells (e.g., mesenchymal stromal cells from the bone marrow, fetal hepatoblasts, embryonic stem [ES] cells), may be used therapeutically for treatment of inherited and acquired liver diseases. This article will summarise our current understanding of these various cell populations, and review possible approaches to their therapeutic use, including cell transplantation, bioartificial liver devices (BLDs), gene therapy and administration of exogenous factors to stimulate normal physiological responses to repair. PMID- 12783610 TI - Bone tissue engineering: recent advances and promising therapeutic agents. AB - Bone regeneration can be accomplished with growth factors, cells and delivery systems. This review is a summary of these components that may be used for tissue regeneration. Support for the potential therapeutic applications of transcription factors in bone tissue engineering will also be discussed. PMID- 12783611 TI - Regeneration therapy for diabetes mellitus. AB - Regeneration therapy can be classified into three categories. The first category, in vitro regeneration therapy, makes use of transplanted cultured cells, including embryonic stem (ES) cells, pancreatic precursor cells and beta-cell lines, in conjunction with immunosuppressive therapy or immunoisolation for the treatment of patients with Type 1 diabetes. In the second type of regeneration therapy, ex vivo regeneration therapy, a patient's own cells, such as bone marrow stem cells, are transiently removed and induced to differentiate into beta-cells in vitro. However, at the present time, insulin-producing cells cannot be generated from bone marrow stem cells. In vivo regeneration therapy, the third type of regeneration therapy, enables impaired tissue to regenerate from a patient's own cells in vivo. beta-Cell neogenesis from non-beta-cells, and beta cell proliferation in vivo have been considered in particular as regeneration therapies for patients with Type 2 diabetes. Regeneration therapy for pancreatic beta-cells can be combined with various other therapeutic strategies, including islet transplantation, cell-based therapy, gene therapy and drug therapy, to promote beta-cell proliferation and neogenesis; it is hoped that these strategies will, in the future, provide a cure for diabetes. PMID- 12783612 TI - Targeting cytokines in autoimmunity: new approaches, new promise. AB - The increasing understanding of the pathophysiology of a number of human autoimmune diseases, and the realisation that cytokines play a major role, has provided the pharmaceutical industry with a wide array of new targets for therapeutic intervention. This has also resulted in a surge of interest for the development of ways of blocking cytokines and their actions in a specific and safe manner. This article reviews the current status of anticytokine therapy and the major efficacy that anti-TNF-a monoclonal antibodies (mAbs) and soluble TNF receptors have demonstrated in the clinic, which has led to their approval for the treatment of rheumatoid arthritis (RA), Crohn's disease (CD), juvenile arthritis and psoriatic arthritis. In addition, the development of novel approaches of cytokine blockade that are based on the characterisation of intracellular signalling pathways regulating cytokine expression (e.g., nuclear factor kappa B [NF-kB] and p38 mitogen activated protein kinase [MAPK]) and the use of small molecule inhibitors are discussed. Whether these approaches will keep up with their early promise and become a major and widespread treatment for several devastating autoimmune diseases will depend on specificity, safety, durability of the benefit, and pharmacoeconomic issues. PMID- 12783613 TI - Non-viral gene transfer therapy for cystic fibrosis. AB - Non-viral methods of gene transfer are being investigated to treat cystic fibrosis (CF) and include naked DNA, lipid-DNA complexes and complexes of DNA with polycations such as poly-L-lysine (poly K) or polyethylenimine (PEI), all of which can carry the cystic fibrosis transmembrane conductance regulator (CFTR) gene. The most recent promising strategy is the use of polycation-DNA complexes, particularly those prepared with poly-K and substituted with polyethylene glycol. These complexes produced partial correction of the CF defect in a mouse model with minimal toxicity, and have advanced to clinical trial. Improvements in this and other non-viral methods are in process and include i). targeting the complexes to the desired cells using receptor ligands, ii). lessening toxicity by changing the mix of lipids or adding protective molecules to polycations, iii). modifying the plasmid DNA to reduce inflammatory CpG sequences and enhance intensity, duration and tissue specificity of expression, and iv). modification of the complexes to improve nuclear access. PMID- 12783614 TI - RNA interference for treating cancers caused by viral infection. AB - The newly discovered phenomenon of RNA interference (RNAi) offers the dual facility of selective viral gene silencing coupled with ease of tailoring to meet genetic variation within the viral genome. Such promise identifies RNAi as an exciting new approach to treat viral-induced diseases, including viral-induced cancers (e.g. cervical carcinoma, hepatocellular carcinomas and haematopoietic and lymphoid malignancies). Cervical cancer is the second most common cancer in women worldwide and is caused by human papilloma virus (HPV). Silencing of HPV gene expression by RNAi induces apoptosis of cervical carcinoma cells in culture, and the effect is apparent within 3 days. The RNAi process is triggered by double stranded RNA (dsRNA), and a single dose is sufficient to maintain RNAi for several days in vitro (cell culture) and in vivo (experimental animal models). Thus, the stage is set for the development of novel antiviral and anticancer therapies based upon selective gene silencing by RNA interference. PMID- 12783615 TI - Therapeutic vaccination to block receptor-ligand interactions. AB - Many chronic diseases are caused by non-physiological interactions of certain ligands with their receptors. Conventional treatment of these diseases with synthetic drugs or monoclonal antibodies (mAbs) is efficient, but problematic due to the non-compliance of patients and the risk of adverse side effects. Novel therapeutic approaches are focusing on strategies of active immunisation aimed at the induction of a humoral immune response directed against the deleterious receptor-ligand interaction. Autoantibody production has been achieved by several vaccine formulations, including conjugates of self-antigens to foreign T helper (Th) cell epitopes, virus-like particles coated with self-antigens, and naked DNA vectors. All of these approaches have the potential to be developed for clinical use if important safety issues, related to the possible long-term presence of self-reactive antibodies in the serum of vaccinated individuals and the risk of undesired T cell responses, can be properly addressed. PMID- 12783616 TI - Therapeutic vaccination for the treatment of mucosotropic human papillomavirus associated disease. AB - There is a high prevalence of diseases caused by human papillomavirus (HPV) infection. Unfortunately, current treatments are inadequate. However, because there is evidence to support a role for the immune system in host defence against this virus, an immunotherapeutic approach is warranted. The existing immunotherapies are not completely effective, nor are they durable. In addition, natural history studies associated with spontaneous regression have provided little guidance to the design of successful interventions. This state of knowledge has encouraged efforts towards the development of novel immunotherapeutic strategies. Successful preclinical studies of therapeutic vaccine candidates have led to clinical studies for a variety of HPV-associated indications, such as anogenital warts and cervical and anal intraepithelial neoplasia. Immunisation approaches such as adjuvanted peptides, virus-like particles and fusion constructs are discussed. Specifically, chimaeric molecules comprised of mycobacterial heat-shock proteins (Hsps) and HPV16 E7 appear promising. PMID- 12783617 TI - Immunotherapy for autoimmune and inflammatory renal diseases. AB - Immune-mediated renal disease (IMRD) accounts for 20 - 30% of the cases of end stage renal failure. It frequently occurs in the context of multi-system autoimmune disorders, including systemic lupus erythematosus (SLE) and primary systemic vasculitis. Current therapies are partially effective and comprise the combination of steroids with an immunosuppressive, such as cyclophosphamide. Their toxicity contributes to the morbidity and mortality of these disorders, and long-term treatment is necessary to prevent relapse. There is a clear need for better-targeted, more effective and less toxic therapy. Advances in our understanding of the immunopathogenesis of inflammatory autoimmune renal disease have identified potential targets for newer agents and have improved the monitoring of therapeutic responses. Recent experience with newer therapies in IMRD is reviewed. This has typically involved small, non-randomised, open-label trials and has addressed reversible features of disease activity. Larger, randomised comparisons to standard therapy are needed along with assessment of long-term efficacy and safety. PMID- 12783618 TI - Darbepoetin alfa: a new approach to the treatment of chemotherapy-induced anaemia. AB - Chemotherapy-induced anaemia has important consequences on the quality of life and social function of cancer patients. The finding of erythropoietin (EPO) deficiency in these patients led to the therapeutic development of erythropoietic proteins. Darbepoetin alfa (Aranesp), Amgen Inc, Thousand Oaks, California), a new erythropoietic growth factor, has eight more sialic acids than epoetin alfa. The increased sialic acid content confers a three-fold longer half-life and allows the drug to be administered less frequently than epoetin alfa. Darbepoetin alfa affects the same early haematopoietic cells as epoetin alfa and the endogenous hormone EPO. Preclinical pharmacokinetic studies suggest that the intrinsic pharmacological properties of darbepoetin alfa are comparable to those of epoetin alfa, but that the increased sialic acid content allows for less frequent administration with superior performance. Darbepoetin alfa has been shown to have safe clinical efficacy in a variety of tumour settings and with several types of chemotherapy. PMID- 12783619 TI - The use of haemoglobin glutamer-250 (HBOC-201) as an oxygen bridge in patients with acute anaemia associated with surgical blood loss. AB - For the treatment of substantial blood loss in surgery, allogeneic blood is transfused to maintain stability and organ perfusion and function. Continued concerns about the availability, safety, efficacy and storage-related problems of allogeneic blood products have led to an intense effort to find alternatives that can serve the same physiologic functions. Haemoglobin-based oxygen carriers (HBOCs) are compounds that can match the oxygen-carrying capacity of red blood cells (RBCs), and several HBOCs have reached advanced stages of development and clinical testing. Multi-centre, randomised, Phase III, controlled trials have demonstrated the safety and efficacy of haemoglobin glutamer-250 (bovine) (Hemopure), Biopure Corporation, Cambridge, MA, USA), also known as HBOC-201. HBOC-201 is bovine-derived, modified haemoglobin that has been ultrapurified to remove any plasma proteins, RBC stroma and potential pathogenic material. During the manufacturing process, crosslinking and polymerisation stabilise the haemoglobin molecule, which increases its vascular persistence as well as the efficiency of oxygen transport to tissue. Results from clinical trials indicate that HBOC-201 can be used as an oxygen 'bridge' for patients experiencing anaemia due to surgical blood loss, until their own red blood cells are replenished or have regenerated (haematinic effect). HBOC-201 is generally well-tolerated and is approved for use in South Africa, where it is indicated for use in adult surgical patients who are acutely anaemic, and is used to eliminate, delay or reduce the need for allogeneic RBCs. A Biologics License Application for HBOC-201 is currently under review by the US FDA. PMID- 12783620 TI - Nasal drug delivery. AB - The Nasal Drug Delivery Conference was held at the Institute of Directors in London, England. The meeting was organised by the Management Forum Ltd and chaired by P Seeney (PA Consulting, UK) and Professor F Merkus (Leiden University, The Netherlands; Innoscience Technology, Belgium). The conference covered a wide range of topics including aspects of nasal physiology, formulation, new nasal products, nasal vaccines, nose to brain transport and pain management via nasal sprays. PMID- 12783621 TI - A reagentless nitric oxide biosensor based on haemoglobin/polyethyleneimine film. AB - An unmediated NO biosensor was prepared by co-immobilizing haemoglobin with polyethyleneimine at a pyrolytic graphite electrode surface. Haemoglobin exhibits direct electrochemistry in the membrane, and this modified electrode shows high stability and catalytic activity towards the reduction of NO. A linear relationship between the catalytic current and the concentration of NO was observed in the concentration ranges 1.0 x 10(-7)-8.0 x 10(-6) M and 1.0 x 10(-5) 2.5 x 10(-5) M, with a detection limit of 4.0 x 10(-8) M. PMID- 12783623 TI - Communicating curriculum reform to students: advice in hindsight... AB - BACKGROUND: In view of the changing health care needs of communities, curriculum reform of traditional curricula is inevitable. In order to allay the apprehension that may accompany such change, curriculum development and implementation should be an inclusive process, with both staff and students being well informed of the planned reform. In 2001, the Nelson R. Mandela School of Medicine implemented Year 1 of a problem-based learning curriculum. During the design phase, students and staff were invited to take part in the development and were kept abreast of developments through meetings and newsletters. METHOD: A survey of Years 1-5 students of the last intake into the traditional curriculum was undertaken a few months prior to the implementation of the new programme. RESULTS: Students were generally well informed about the impending change, having heard about it from fellow students and staff. The more senior the students, the less the perceived impact of the reform. Although most of what students had heard was correct, some, however, had misconceptions that were generally extreme views (e.g. all self directed learning; no Anatomy) about the new programme. Others expressed valid concerns (e.g. underpreparedness of students from disadvantaged schools; overcrowding in hospitals). CONCLUSIONS: Advice offered to institutions considering curriculum reform include using various methods to inform internal and external affected parties, ensuring that the student representative body and staff is well informed, reiterating the need for the change, confirming that the new programme meets recognised standards and that the students most affected are reassured about their future studies. PMID- 12783622 TI - Measuring the lung function in the mouse: the challenge of size. AB - Measurement of the effects of drugs, mediators and infectious agents on various models of lung disease, as well as assessment of lung function in the intact mouse has the potential for significantly advancing our knowledge of lung disease. However, the small size of the mouse presents significant challenges for the assessment of lung function. Because of compromises made between precision and noninvasiveness, data obtained may have an uncertain bearing on the mechanical response of the lung. Nevertheless, considerable recent progress has been made in developing valid and useful measures of mouse lung function. These advances, resulting in our current ability to measure sophisticated indices of lung function in laboratory animals, are likely to lead to important insights into the mechanisms of lung disease. PMID- 12783624 TI - Feasibility and effectiveness of offering a solution-focused follow-up to employees with psychological problems or muscle skeletal pain: a randomised controlled trial. AB - BACKGROUND: Long-term sick leave has been of concern to politicians and decision makers in Norway for several years. In the current study we assess the feasibility and effectiveness of offering a voluntary, solution-focused follow-up to sick-listed employees. METHODS: Employees on long-term sick leave due to psychological problems or muscle skeletal pain were randomly allocated to be offered a solution-focused follow-up (n = 122) or "treatment as usual" (n = 106). The intervention was integrated within 2 social security offices' regular follow up. The intervention group was informed about the offer with letters, telephone calls and information meetings. Feasibility was measured by rate of uptake to the intervention, and effectiveness by number of days on sick leave. RESULTS: In general, few were reached with the different information elements. While the letter was sent to all, only 31% were reached by telephone and 15% attended the information meetings. Thirteen employees (11.5%) in the intervention group participated in the solution-focused follow-up. Intention to treat analysis showed no difference in mean length of sick leave between the intervention group (217 days) and the control group (189 days) (p = 0,101). CONCLUSION: Even if the information strategy might be improved, it is not likely that a voluntary solution-focused follow-up offered by the social security offices would result in measurable reduction in length of sick leave on a population level. However, the efficacy of a solution-focused follow-up for the persons reporting a need for this approach should be further investigated. PMID- 12783625 TI - The rat STSL locus: characterization, chromosomal assignment, and genetic variations in sitosterolemic hypertensive rats. AB - BACKGROUND: Elevated plant sterol accumulation has been reported in the spontaneously hypertensive rat (SHR), the stroke-prone spontaneously hypertensive rat (SHRSP) and the Wistar-Kyoto (WKY) rat. Additionally, a blood pressure quantitative trait locus (QTL) has been mapped to rat chromosome 6 in a New Zealand genetically hypertensive rat strain (GH rat). ABCG5 and ABCG8 (encoding sterolin-1 and sterolin-2 respectively) have been shown to be responsible for causing sitosterolemia in humans. These genes are organized in a head-to-head configuration at the STSL locus on human chromosome 2p21. METHODS: To investigate whether mutations in Abcg5 or Abcg8 exist in SHR, SHRSP, WKY and GH rats, we initiated a systematic search for the genetic variation in coding and non-coding region of Abcg5 and Abcg8 genes in these strains. We isolated the rat cDNAs for these genes and characterized the genomic structure and tissue expression patterns, using standard molecular biology techniques and FISH for chromosomal assignments. RESULTS: Both rat Abcg5 and Abcg8 genes map to chromosome band 6q12. These genes span ~40 kb and contain 13 exons and 12 introns each, in a pattern identical to that of the STSL loci in mouse and man. Both Abcg5 and Abcg8 were expressed only in liver and intestine. Analyses of DNA from SHR, SHRSP, GH, WKY, Wistar, Wistar King A (WKA) and Brown Norway (BN) rat strains revealed a homozygous G to T substitution at nucleotide 1754, resulting in the coding change Gly583Cys in sterolin-1 only in rats that are both sitosterolemic and hypertensive (SHR, SHRSP and WKY). CONCLUSIONS: The rat STSL locus maps to chromosome 6q12. A non-synonymous mutation in Abcg5, Gly583Cys, results in sitosterolemia in rat strains that are also hypertensive (WKY, SHR and SHRSP). Those rat strains that are hypertensive, but not sitosterolemic (e.g. GH rat) do not have mutations in Abcg5 or Abcg8. This mutation allows for expression and apparent apical targeting of Abcg5 protein in the intestine. These rat strains may therefore allow us to study the pathophysiological mechanisms involved in the human disease of sitosterolemia. PMID- 12783627 TI - Rapid de-localization of actin leading edge components with BDM treatment. AB - BACKGROUND: 2,3-butanedione monoxime (BDM) has been widely used as a non-muscle myosin inhibitor to investigate the role of non-muscle myosinII in the process of actin retrograde flow and other actin cytoskeletal processes. Recent reports show that BDM does not inhibit any non-muscle myosins so far tested, including nm myosinII, prompting the question, how were these process affected in BDM studies? RESULTS: We have found that treatment of mammalian cells with BDM for only 1 min blocks actin incorporation at the leading edge in a permeabilized cell system. We show that inhibition of actin incorporation occurs through de-localization of leading edge proteins involved in actin polymerization--the Arp2/3 complex, WAVE, and VASP--that de-localize concomitantly with the leading edge actin network. CONCLUSION: De-localization of actin leading edge components by BDM treatment is a newly described effect of this compound. It may explain many of the results previously ascribed to inhibition of non-muscle myosinII by BDM, particularly in studies of leading edge dynamics. Though this effect of BDM is intriguing, future studies probing actin dynamics at the leading edge should use more potent and specific inhibitors. PMID- 12783629 TI - ALLHAT: is the final answer in? PMID- 12783628 TI - EasyGene--a prokaryotic gene finder that ranks ORFs by statistical significance. AB - BACKGROUND: Contrary to other areas of sequence analysis, a measure of statistical significance of a putative gene has not been devised to help in discriminating real genes from the masses of random Open Reading Frames (ORFs) in prokaryotic genomes. Therefore, many genomes have too many short ORFs annotated as genes. RESULTS: In this paper, we present a new automated gene-finding method, EasyGene, which estimates the statistical significance of a predicted gene. The gene finder is based on a hidden Markov model (HMM) that is automatically estimated for a new genome. Using extensions of similarities in Swiss-Prot, a high quality training set of genes is automatically extracted from the genome and used to estimate the HMM. Putative genes are then scored with the HMM, and based on score and length of an ORF, the statistical significance is calculated. The measure of statistical significance for an ORF is the expected number of ORFs in one megabase of random sequence at the same significance level or better, where the random sequence has the same statistics as the genome in the sense of a third order Markov chain. CONCLUSIONS: The result is a flexible gene finder whose overall performance matches or exceeds other methods. The entire pipeline of computer processing from the raw input of a genome or set of contigs to a list of putative genes with significance is automated, making it easy to apply EasyGene to newly sequenced organisms. EasyGene with pre-trained models can be accessed at http://www.cbs.dtu.dk/services/EasyGene. PMID- 12783630 TI - Pharmacokinetics of propranolol after single and multiple dosing with sustained release propranolol or propranolol CR (innopran XL) , a new chronotherapeutic formulation. AB - Blood pressure rises rapidly upon waking and may be responsible, in part, for the increased incidence of myocardial infarction and stroke during the morning hours. Current formulations and dosing of antihypertensive drugs do not provide maximum coverage during this vulnerable period. This study was performed to demonstrate that propranolol CR (Innopran XL), a novel chronotherapeutic formulation of propranolol designed for nighttime dosing, has appropriate pharmacokinetics to provide maximum cardioprotective effect in the morning. Pharmacokinetics of propranolol CR and sustained-release propranolol after single and multiple doses were determined in normal male volunteers in this open-label, 2-period crossover study. The drugs were dosed in the evening and serial blood samples were taken for determination of propranolol concentration the next 24 to 72 hours. After a single 160-mg dose of propranolol CR administered at 10 pm, absorption was delayed by about 4 hours, after which plasma concentration rose steadily, reaching a peak at about 10:00 am. In contrast, after dosing with sustained release propranolol, plasma levels of propranolol began to rise almost immediately, reaching a plateau between 4:00 am and 10:00 am. During multiple dosing, steady-state trough plasma concentrations were achieved after 2 days with either drug. After the final dose, the plasma profiles of both drugs were similar to those observed in the single-dose study. Bioavailability was similar for both formulations of propranolol. Propranolol CR exhibited appropriate pharmacokinetics for a chronotherapeutic approach to the treatment of hypertension. PMID- 12783631 TI - Inflammation at a glance: erythrocyte adhesiveness/aggregation test to reveal the presence of inflammation in people with atherothrombosis. AB - The erythrocyte adhesiveness/aggregation test is a new biomarker to detect low grade inflammation in patients with atherothrombosis. In a group of 30 individuals with an acute ischemic event, the variability of EAAT during a follow up period was similar to those obtained for other commonly used markers of the acute phase response, suggesting the potential clinical use of this novel marker. PMID- 12783632 TI - Circumflex coronary artery occlusion after blunt chest trauma. AB - A 32-year-old white male police officer suffered blunt trauma to the anterior chest wall during a routine training session. This was accompanied by the precipitous onset of chest discomfort. There was no previous history of any cardiac risk factors. The diagnosis of an inferior wall myocardial infarction was made based on the electrocardiogram findings, at his local community hospital. The total creatine kinase, creatine kinase-MB, and troponin I were normal. The transesophageal echocardiogram performed at that time demonstrated no aortic or coronary dissection. He was transferred to our tertiary care center. Emergency cardiac catheterization demonstrated lateral wall hypokinesis with a left ventricular ejection fraction of 45% and a total occlusion of the left circumflex coronary artery in its proximal portion. This was successfully recannulized with angioplasty and stenting techniques. We believe this to be only the second reported case of circumflex coronary artery obstruction after blunt chest trauma. PMID- 12783626 TI - Gene discovery in the hamster: a comparative genomics approach for gene annotation by sequencing of hamster testis cDNAs. AB - BACKGROUND: Complete genome annotation will likely be achieved through a combination of computer-based analysis of available genome sequences combined with direct experimental characterization of expressed regions of individual genomes. We have utilized a comparative genomics approach involving the sequencing of randomly selected hamster testis cDNAs to begin to identify genes not previously annotated on the human, mouse, rat and Fugu (pufferfish) genomes. RESULTS: 735 distinct sequences were analyzed for their relatedness to known sequences in public databases. Eight of these sequences were derived from previously unidentified genes and expression of these genes in testis was confirmed by Northern blotting. The genomic locations of each sequence were mapped in human, mouse, rat and pufferfish, where applicable, and the structure of their cognate genes was derived using computer-based predictions, genomic comparisons and analysis of uncharacterized cDNA sequences from human and macaque. CONCLUSION: The use of a comparative genomics approach resulted in the identification of eight cDNAs that correspond to previously uncharacterized genes in the human genome. The proteins encoded by these genes included a new member of the kinesin superfamily, a SET/MYND-domain protein, and six proteins for which no specific function could be predicted. Each gene was expressed primarily in testis, suggesting that they may play roles in the development and/or function of testicular cells. PMID- 12783633 TI - Cardiovascular manifestations of substance abuse part 1: cocaine. AB - Substance abuse with cocaine is associated with multiple cardiovascular conditions, including myocardial infarction, dissection, left ventricular hypertrophy, arrhythmias, sudden death, and cardiomyopathy. Cocaine has effects to potentiate the physiologic actions of catecholamines and has direct effects on voltage-dependent sodium ion channels related to local anesthetic properties. The effects of cocaine can be augmented with concomitant alcohol consumption. Acute myocardial ischemia caused by cocaine may be related to in situ thromboisis and/or coronary vasospasm. Treatment strategies for cocaine-induced myocardial infarction would include antiplatelet therapy, thrombolysis, and vasodilators (eg, nitrates, nifedipine). Beta-adrenergic blockers should not be used unless concomitant vasodilator therapy is given. PMID- 12783634 TI - Gender, ethnicity, and genes in cardiovascular disease. Part 2: implications for pharmacotherapy. AB - Women are underrepresented in clinical trials. Lower doses of beta-blockers are required for Southeast Asians. ACE and ARB's are teratogenic in the second trimester. Torsades de Pointes is more common in women related to a longer QT interval. Lower dose OCPs decrease the risk of MI, stroke and thrombosis. HRTs are not effective for CAD prevention. PMID- 12783635 TI - Natriuretic peptides: biochemistry, physiology, and therapeutic role in heart failure. AB - Cardiac natriuretic peptides are a family of structurally related peptides that are important in sodium and volume homeostasis. They consist of atrial natriuretic peptide, brain natriuretic peptide, and C-type natriuretic peptide and are elevated in patients with left ventricular dysfunction. In contrast with vasoconstrictive hormones, such as norepinephrine, angiotensin II, and arginine vasopressin, which worsen the physiological milieu in patients with left ventricular dysfunction and heart failure, the natriuretic peptides ameliorate these effects by promoting natriuresis, diuresis, peripheral vasodilation, and by inhibiting the renin-angiotensin system. The serum levels of the natriuretic peptides correlate with the severity of heart failure and appear to have prognostic value. The present article reviews the biochemistry, molecular biology, and physiology of natriuretic peptides and their pathophysiological link to heart failure. The therapeutic uses of natriuretic peptides are also reviewed. This includes the use of intravenous nesiritide, a synthetic human brain natriuretic peptide, and the recently developed vasopeptidase inhibitors which are designed to inhibit the degradation of natriuretic peptides. PMID- 12783636 TI - Atrial myxoma: case report and a review of the literature. AB - Atrial myxomas are the most common benign primary tumor of the heart and occur in as many as 3 in 1000 patients. These tumors are a major cause of patient morbidity and mortality. Although the majority of atrial myxomas occur in the left atrium, 3 separate familial myxoma syndromes can result in multiple myxomas in atypical locations. Approximately 50% of patients with myxomas may experience symptoms due to central or peripheral embolism or intracardiac obstruction, but 10% of patients may be completely asymptomatic. Screening for myxomas should involve a thorough history and physical examination and a transthoracic and/or transesophageal echocardiogram. Transthoracic echocardiography is approximately 95% sensitive for the detection of cardiac myxomas, and transesophageal echocardiography approaches 100% sensitivity. Though the majority of atrial myxomas are sporadic, it is imperative that first-degree relatives of patients with documented myxomas undergo screening for occult myxomas. Surgical removal of the myxoma is the treatment of choice and usually curative; however, myxoma recurrence does occur and is most frequently associated with a familial syndrome. PMID- 12783637 TI - Insulin glargine: a new once-daily basal insulin for the management of type 1 and type 2 diabetes mellitus. AB - Optimal glucose control is the primary goal for treating diabetes mellitus and preventing long-term complications of diabetes, such as coronary heart disease, nephropathy, neuropathy and retinopathy. Insulin glargine is a novel, long-acting human insulin analog that is indicated in type 1 diabetic patients aged >or=6, or in type 2 diabetic patients who require basal insulin for glycemic control. Insulin glargine is a recombinant insulin with a modified structure that allows it to dissolve in an acidic solution, but to precipitate in the physiological subcutaneous tissue forming a depot effect. In contrast to Neutral Protamine Hagedorn (NPH) insulin, insulin glargine has a slower onset, a longer duration of action, and no peak in metabolic activity. Once-daily subcutaneous administration of insulin glargine at bedtime has comparable efficacy to that of NPH insulin once or twice daily when used in combination with bolus insulin in type 1 diabetic patients, or in conjunction with oral antidiabetic drugs in type 2 diabetic patients. Overall, insulin glargine has similar adverse effects when compared with NPH insulin. Insulin glargine has been associated with less nocturnal hypoglycemia and improved treatment satisfaction in several clinical trials with durations of < 52 weeks. Pharmacoeconomic analysis comparing insulin glargine with other intermediate- or long-acting insulin preparations used as basal insulin therapy has not been performed. In summary, insulin glargine offers a promising alternative as a once-daily basal insulin therapy in patients with type 1 and type 2 diabetes. PMID- 12783638 TI - The effect of δ-opioid agonists on intracellular calcium level in MOLT-4 T cell line. AB - δ-opioid agonists were reported to affect T cell functions: proliferation, cytotoxicity, cytokine production. Changes in intracellular calcium level are important in T-cell activation. In this study the effect of the synthetic δ-opioid agonist DADLE and an endogenous δ-opioid pentapeptide Met enkephalin on the intracellular calcium level in human T-lymphoblastic leukemia MOLT-4 cells is reported. Intracellular calcium level was monitored using QUIN 2 AM as a fluorescent dye in MOLT-4 cells after short treatment (2 and 15 min) with DADLE and Met-enkephalin. DADLE (10-8M) mildly (average 28%) decreased the intracellular calcium level after 1 min treatment. The suppressive effect of DADLE (10-8M) on the intracellular calcium level was enhanced by longer (15 min) treatment of MOLT-4 cells (average 40%). Met enkephalin (10-9M - 10-7M) decreased (average 33 %) the intracellular calcium level after 2 min treatment (average 33% - 37%). However, Met-enkephalin (10-7M) increased (average 31%) the intracellular calcium level after longer (15 min) treatment. Ionophore A23187 (10-7M, 10-6M) was used as a positive control to enhance intracellular calcium level. Thus, δ-opioid agonist DADLE decreased basal intracellular calcium level in MOLT-4 cells after short treatment, while endogenous Met-enkephalin altered intracellular calcium level in a bidirectional way by decreasing and increasing it. PMID- 12783639 TI - Hepatitis C virus infection is frequently associated with high titers of anti thyroid antibodies. AB - Objective: To assess the implication of hepatitis C virus (HCV) infection in the development of autoimmune thyroid response, thyroid autoantibodies were studied in serum of HCV positive patients. Methods: Anti-microsomal, anti-thyroperoxidase (anti-TPO) and anti-thyroglobulin antibodies were evaluated in the sera of 100 patients with chronic hepatitis C (53 women and 47 men; mean age = 55±5 years). In parallel, thyroid autoantibodies were investigated in blood samples obtained from two separated control groups: age-matched HCV negative-HBV positive patients (25 women, 25 men; mean age= 47±6 years) and healthy blood donors (29 women and 21 men; mean age= 54±8 years). Results: Anti-thyroid antibodies were found more frequently in HCV positive women when compared to the men (8/53= 15.1% vs 0/47= 0%,.p<0.01).The prevalence of these autoantibodies was not statistically different between HCV positive and healthy female blood donors. However the investigation of thyroid autoantibody titers showed significantly higher levels of anti-TPO and anti-microsomal antibodies in HCV positive women in comparison with healthy women controls (respectively 1: 83200 vs 1: 1900 and 834 vs 23, p<0,01). Conclusions: This strong association between HCV infection and high levels of anti-thyroid autoantibodies in women outlines the interest of systematic detection of anti-microsomal and/or anti-TPO antibodies in this population. PMID- 12783640 TI - Myasthenia gravis: correlation of age with clinical course and anti-AChr antibody levels. AB - We studied the significance of age as a factor affecting prognosis and anti-AChR antibody levels in myasthenia gravis, comparing patients with symptom onset before 50 (37 patients) and at or after 50 (33 patients). More of the older patients, compared to younger ones (69.9% versus 32.4%,p<0. 05) progressed to severe disease in the first three years of treatment. Five older patients died during myasthenic crises, whereas no deaths occured in the younger group. In addition; older patients did not respond as favorably as younger ones to anticholinesterase treatment and more of them required combination therapy. AntiAChR antibody levels were significantly lower in older patients (9.2 versus 34.9 nM, p<0.005). We conclude that symptom onset at or after the age of 50 predicts unfavorable outcome in myasthenia gravis and is associated with lower anti-AChR antibody levels. This group of patients requires earlier treatment with corticosteroids, immunosuppressive drugs or both. PMID- 12783641 TI - Dry eye in allergic conjunctivitis: role of inflammatory infiltrate. AB - Dry-eye associated symptoms are frequently present in patients affected with allergic conjunctivitis. We evaluated the relationship between ocular inflammation and the tear film parameters in patients affected with chronic allergic conjunctivitis. Eighty-two subjects (age 23.00 ± 7.61: range 10 40) affected with chronic allergic conjunctivitis and thirty age- and sex matched healthy controls were enrolled. In all patients tear film qualitative and quantitative tests were performed. To determine the immune activation state, conjunctival biopsies were obtained from the inferior conjunctival fornix. Immunocytochemical markers for CD45RO, CD8, CD20 and EG2 (monoclonal antibody binding eosinophil cationic protein) were evaluated semiquantitatively. All tear film tests were found altered. They were reduced in allergic patients (p<0.001). In conjunctival biopsies of allergic patients a very high number of CD45RO+ and EG2+ cells were found (p<0.001): a lower number of CD45RO+ cells and no EG2+ cells in control biopsies were found. Multivariate analysis showed a significant relationship between tear tests and conjunctival infiltrate (CD45RO+ and EG2+): The tear film alterations are strictly related to the conjunctival immune infiltration. In particular, the reduction of the mucin related component of tear film can be related to the toxic effect of the granule cationic proteins released by the conjunctival activated eosinophils (EG2+ cells). PMID- 12783642 TI - Leukocyte adhesion deficiency disorder in an infant. AB - Leukocyte adhesion defect (LAD) is an inherited disorder of phagocytic function. The three different alpha-chains of the leukocyte integrin family (CD 11a, 11b, 11c) are all dependent on the common beta chain (CD18) for proper insertion into the cell membrane. Deficiency of CD 18 thus causes an inability to express any of these proteins and results in the syndrome of LAD type 1. Clinical features include delayed separation of the umbilical cord, persistent leukocytosis and recurrent infection. A case of severe LAD occurred in a 1 month-old infant. The immunological pattern of the baby showed pathological values of CD11b and CD18 (1.3% and 0.2% respectively) when compared with normal values (50 70%) and lack of random (1.0 nm vs normal of 10±5) and casein induced (22.0 nm vs normal of 60-120nm) chemotaxis. The baby is now waiting for bone marrow transplantation. Although LAD is a rare form of congenital immune deficiency this disease should be considered when delayed wound healing and recurrent bacterial skin infections are present in a newborn. PMID- 12783643 TI - Invasion of human cultured cells by Prevotella nigrescens. AB - Invasion of gingival and junctional epithelial cells has been recently proposed as a potentially relevant mechanism in the pathogenesis and recurrence of periodontal disease. The gram negative anaerobe Prevotella nigrescens was shown to be involved in the development of periodontal lesions in man, suggesting a possible involvement of invasivity as a mean to circumvent the host immune surveillance and other hostile factors. Appropriately designed invasion assays demonstrated that P. nigrescens efficiently invades human epithelial cells, through a mechanism whose efficiency is influenced by the phase of growth, by the multiplicity of infection, and by the cell line used, and that requires microfilament integrity, but is not affected by an impairment of microtubule organization. Intracellular replication assays suggested that P. nigrescens probably multiplies within Kb epithelial cells, causing extensive cell alterations. Invasion of gingival epithelial cells could consequently be a basic step in the virulence mechanism of the species. PMID- 12783644 TI - The anti-invasive effect of bovine lactoferrin requires an interaction with surface proteins of Listeria Monocytogenes. AB - The anti-invasive effect of bovine lactoferrin (BLf) and of bovine transferrin (BTf) towards L. monocytogenes, an intracellular facultative food-borne pathogen, was assayed in the enterocyte-like cell line Caco-2. When 0.5 mg/ml BLf were added during the infection time or preincubated with bacteria the number of internalized bacteria was noticeably decreased whereas BLf was ineffective when preincubated with the enterocytes or added post infection. BTf was deprived of any effect. Results from direct binding and Western blotting assays provided evidence that two L. monocytogenes surface proteins, of approximately 80 and 60 kDa, specifically reacted with BLf. These findings strongly support the hypothesis that the antiinvasive mechanism of BLf is due to its interaction with bacterial surfaces, but not to its binding with eukaryotic cells. PMID- 12783645 TI - Flunitrazepam a benzodiazepine most used among drug abusers. AB - Flunitrazepam (FZ) is a sedative/hypnotic nitro-benzodiazepine. This drug has been accepted by both patients and physicians and in the last 20 years flunitrazepam has been included and studied in many clinical trials so, in many countries, flunitrazepam is one of the most prescribed hypnotic. Since 1980 it has been found that FZ began to be a popular drug among drug abusers all over the word. However, little is known about the difference between Fz and other Benzodiazepines in capacity to produce physiologic dependence or in ability to produce drug taking or drug seeking behaviour. Flunitrazepam has little risk of abuse by the vast majority of patients; however when the drug is taken i. v. or intranasally, its effect is much faster and risk of abuse is much greater. In this report we examine the reasons why some populations of drug abusers prefer flunitrazepam over the other benzodiazepines. PMID- 12783646 TI - Increased expression of intercellular adhesion molecule 1 in relation to mast cells in the bladder of interstitial cystitis patients. AB - Interstitial cystitis (IC) is a sterile, painful bladder disorder, often associated with inflammation. A defective urothelium and activated mast cells have been implicated in the pathophysiology of IC. Mast cells have been reported to induce the expression of intercellular adhesion molecules (ICAM) that are considered a necessary first step for the initiation of inflammation. We investigated the presence of ICAM-1 and its association with bladder mast cells in IC patients and controls in order to further understand their role in the pathophysiology of IC. Bladder tissue was fresh frozen and 7m section were cut using a cryostat. Immunocytochemical detection was carried out with monoclonal antibodies directed against anti-ICAM1 and tryptase. Twenty-one biopsies from women with IC and two from normal tissue from radical cystectomies (control) were analyzed. All IC samples were strongly positive for ICAM-1, exclusively in capillaries. Control samples were only faintly positive in fewer capillaries. Numerous mast cells were identified in the positive regions with toluidine blue or tryptase immunocytochemistry. This work may help explain why many IC patients develop sterile inflammation. It may also provide evidence further explaining the role of bladder mast cells in the pathophysiology of IC. PMID- 12783647 TI - A Th2-like cytokine response is involved in bullous pemphigoid. the role of IL-4 and IL-5 in the pathogenesis of the disease. AB - Bullous Pemphigoid is an autoimmune bullous disorder characterized by production of IgG against an hemidesmosomal antigen (230 kDa, 180 kDa) responsible for blistering of the skin. In the past several mediators have been implicated in the pathogenesis of the disease such as proteases and collagenases secreted by local inflammatory cells. In order to investigate the role of cytokines in BP, the cytokine pattern was evaluated by an immunohistochemical analysis and by reverse transcriptase polymerase chain reaction procedure in 13 BP patients. Cytokines examined were interleukin (IL)-2, IL-4, IL-5, interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha. The T cell inflammatory infiltrate was also characterized by monoclonal antibodies showing CD3+, CD4+ T cells with a perivascular and scattered distribution in lesional skin. IL-4 and IL-5 were detected in a similar distribution to the inflammatory infiltrate. IL-4 and IL-5 mRNA levels were also revealed by RT-PCR. Proinflammatory cytokines such as TNF alpha, IL-6 and Th1-like cytokines (IL-2 and INF-gamma) were not detected neither as proteins nor as mRNA. Since IL-4 and IL-5 are important in eosinophil chemoattraction, maturation and functional activity, the presence of IL-4 and IL 5 in BP suggest that these cytokines could be important in the pathogenesis of the disease. PMID- 12783648 TI - Immune response in liver transplantation: is there a preferential pattern in acute rejection? AB - To determine the immune factors involved in liver graft rejection, a study on 14 liver transplants was conducted. We have, in particular, studied CD4+CD7+ and CD8+CD38+ T cells in both liver tissue and blood of patients with and without acute rejection. Contemporarily, IL-2 and IL-4 secretion in both plasma and stimulated culture supernatants from hepatic T cells was evaluated. Early acute rejection was characterized by a higher expression of CD4+CD7+ and CD8+CD38+ T lymphocytes in the liver than in blood (p<0.001). Moreover, a preferential proinflammatory (Thl) cytokine profile was related to liver resident T cells in comparison with corresponding plasma (p<0.001). Conversely, in the patients without acute rejection CD4+CD7+ was higher in blood than in liver and the Th2-like cytokine profile characterized these subjects. Our data suggest that a preferential Th1 immune mechanism operates in a local fashion and may be involved in acute rejection. PMID- 12783649 TI - Nitric oxide dependent killing of mycobacterium tuberculosis by human mononuclear phagocytes from patients with active tuberculosis. AB - In this study we have demonstrated that nitric oxide, the product of the arginine dependent pathway of human mononuclear phagocytes effectively kills the M.tuberculosis in-vitro. The release of reactive nitrogen intermediates was triggered by incubation with various proinflammatory cytokines namely IFN gamma,TNF-alpha and IL-1R. We have earlier shown that human mononuclear phagocytes can be induced to release nitric,oxide (NO) radicals which can kill tumour cells. In the present communication, by using colony forming assays we demonstrated that human mononuclear phagocytes can effectively kill M.tuberculosis by using a NO dependent pathway. Treatment of mononuclear phagocytes with L-arginine resulted in markedly increased killing activity whereas, by using NGMMA, an analogue of L-arginine, the cidal activity could be brought down to the basal level. These results clearly suggest that cytokines, particularly IFN-gamma, induced NO release and its reactive product with oxygen radical, peroxynitrite, could play an important role in the killing of M. tuberculosis by human mononuclear phagocytes. A significant production of interleukin-4 and interleukin-10, by the ex-vivo matured, untreated macrophages from the active tuberculosis patients indicate that regulation of cytokine network to encourage in situ/local production of nitric oxide may be useful in the management of pulmonary tuberculosis. PMID- 12783650 TI - Evaluation of risk and benefit of oral immunomodulation using heat-killed enterococcus faecalis FK-23 preparation in healthy dogs. AB - The risk and benefit of oral immunomodulation using heat-killed Enterococcus faecalis FK-23 preparation (FK-23) were evaluated in healthy dogs. Dogs were administered perorally 100 mg/kg of FK23 daily for 3 months. FK-23 did not affect the findings of complete blood count, leukocyte differential count, blood chemistry, serum electrolyte, radiography, or physical conditions of the dogs tested. Increase in myeloid/erythroid ratio and granulocytic lineage was found in bone marrow of the FK-23 treated dogs. Neutrophil function assessed by zymosan dependent chemiluminescence was activated by treatment of the drug. FK-23 also stimulated in vitro lymphocyte blast transformation of PHA, Con A, and LPS. FK-23 thus appears to be a useful immunomodulating drug to stimulate non-specific host immune responses without adverse side effects. PMID- 12783651 TI - Virulence factors of salmonella ser. enteritidis strains isolated in Italy from food-borne outbreaks. AB - Twelve Salmonella ser. Enteritidis strains phage type 4 isolated in Italy from different food-borne outbreaks were characterized for the expression of different virulence traits, for antibiotic resistance, and for plasmid DNA profile. All the twelve S. Enteritidis strains were able to invade and multiply within HeLa cell monolayers, even if at a lower efficiency if compared to an invasive Shigella flexneri strain. The strains were not hemolytic and produced only a moderate level cytotoxic effect on HeLa cell monolayers. Moreover, all the strains examined produced mannose-sensitive hemagglutination with chicken erythrocytes but were not able to adhere to tissue culture cells. The strains did not produce the hydroxamate-type siderophore aerobactin or the specific ferric-aerobactin receptor. The S. Enteritidis strains were resistant only to spectinomycin, and eleven strains harbored a 38 MDa non-conjugative plasmid, while one strain harbored a 64 MDa conjugative plasmid which carried a colicinogenic activity encoding locus. The uniformity of antibiotic resistance pattern, of the plasmid DNA content, and of the virulence factors produced indicated that the S. Enteritidis clinical isolates examined are clonally-related. PMID- 12783652 TI - Influence of the oral administration of lactic acid bacteria on iga producing cells associated to bronchus. AB - Intestinal, respiratory and genitourinary mucosal surfaces are the most important routes of entry for microbial pathogens. The stimulus of the mucosal immunity is not easy because the trigger keys for the activation do not follow the ones of the systemic immune response. In previous works we have demonstrated that some Lactic Acid Bacteria (LAB), when orally administered, can induce an enhance of the gut immune response. Taking into account the concept of a common mucosal response, we studied the effect of orally administered mice with Lactobacillus casei, L. acidophilus, L. rhamnosus, L. delbrueckii subsp. bulgaricus, Streptococcus salivarius subsp. thermophilus and Lactococcus lactis on the IgA secreting cells associated to bronchus. As shown before, oral immunostimulation with LAB induced an increase of the IgA* cells at intestinal level by a dose depending effect. In this study it is also showed that the LAB assayed, with exception of L. acidophilus, were able to enhance IgA+ cells at bronchial level, being also this effect dose dependent. The increment induced by some LAB in the number of IgA+ cells on the mucosa surface of the lower respiratory tract may be very important to prevent bronchus diseases. PMID- 12783653 TI - Effect of bacterial extracts on the immunologic profile in chronic relapsing brucellosis patients. AB - Brucellosis is an intracellular bacterial disease of common incidence in Greece. Existing therapy is inadequate and a considerable proportion of patients become chronically ill and are immunocompromised. Defects of the monocyte-macrophage system and T-lymphocytes have been described in chronic brucellosis and can be restored after immunopotentiation therapy. Bacterial (Klebsiella pneumoniae) extracts exert immunostimulating effects on the monocyte-macrophage system and have already been used successfully in the prevention of common infections of the respiratory track. So we decided to investigate: 1) Leukocyte Migration Index (LMI), 2) Monocyte-macrophage random and directed migration against both nonspecific leukoattractant (casein) and disease specific antigens (Brucella melitensis, Brucella abortus), 3) Monocyte-macrophage phagocytosis index, 4) Delayed-type hypersensitivity (skin tests) against seven antigens, before (TO), during (T2), and after (T3) oral administration of bacterial (Klebsiella pneumoniae) extracts at conventional doses plus antibiotics or not. Our results show that: 1) Concerning the LMI, 4 out of 19 remained anergic at time T3 of the study, 2) Random migration was not affected during treatment, 3) Directed migration increased significantly without reaching control group values, 4) Phagocytosis index increased significantly and reached normal values at T3, 5) Delayed type hypersensitivity reactions (skin tests) increased significantly at the end of the study period. Reaction against Tuberculin and Candida antigens showed the most pronounced increase in skin reactivity. In conclusion, bacterial (Klebsiella pneumoniae) extracts improve peripheral monocyte locomotion and restore phagocytosis index, thus enhancing cellular immunity parameters in immunocompromised chronic brucellosis patients. PMID- 12783654 TI - [The short-term and long-term treatment outcomes in patients with pulmonary tuberculosis positive for drug-resistant and sensitive strains]. AB - OBJECTIVES: To analyze the treatment outcomes in patients with smear positive tuberculosis, and to compare the difference in treatment response among patients infected with drug-sensitive and drug-resistant strains. METHODS: From 1998 to 2000, seven hundred and seventy-seven patients with primary smear-positive tuberculosis, which were from 30 surveillance sites, were followed for two years to monitor their treatment outcomes. RESULTS: At the completion of the 6 months' therapy, the overall rate of treatment failure was 1.8%, 2.6% for the drug resistant cases and 1.6% for the drug-sensitive cases. Six-month follow-up showed a positive conversion rate of 2.7% in all the cases, 8.5% and 1.2% (P < 0.005) in the drug-resistant and the drug-sensitive cases respectively. One year follow-up showed that the positive conversion rate was 2.6% in all the cases, 6.9% and 1.6% (P < 0.005) in the drug-resistant and the drug-sensitive cases, respectively. Two year follow-up showed an overall positive conversion rate of 1.3%, 1.0% and 1.3% in the drug-resistant and the drug-sensitive cases, respectively. Of the 152 drug resistant cases, the rate of treatment failure was 2.6% at the completion of 6 months' therapy, but in cases with MDR-TB the rate was 10.3%. Six-month follow-up showed an overall positive conversion rate of 8.5%, but the rate reached 37.0% in cases with MDR-TB. One-year and two-year follow-up showed that the positive conversion rates were 6.9% and 1.0% respectively in all the drug-resistant cases, but 6.3% and 6.7% respectively in the MDR cases. CONCLUSIONS: Under the guidelines of the National Tuberculosis Program (NTP), the 2H(3)R(3)S(3)Z(3)/4H(3)R(3) regimen for primary smear-positive pulmonary TB was effective. But the cure rate was lower and the positive conversion rate higher in patients with MDR-TB. PMID- 12783655 TI - [Follow-up observation on relapse of smear negative pulmonary tuberculosis after short-course chemotherapy]. AB - OBJECTIVES: To evaluate the efficacy of the tuberculosis (TB) control project supported by WPRO/WHO in Jiangsu province, China, and to analyze the factors associated with relapse. METHODS: All registered patients with smear positive pulmonary tuberculosis which were diagnosed and cured in the project, were examined for bacteriological relapse after 2 to 3 years. The relationships among relapse rate, bacterial load and the time of sputum negative conversion were analyzed by historical cohort study. RESULTS: Fifty seven cases of 1,730 patients who were followed for 2 - 3 years were found smear positive, the relapse rate being 3.29%. The relapse rates in patients whose sputum became negative within two months, within three months and beyond three months after the start of therapy were 2.1%, 5.3% and 8.8% (P < 0.01), respectively. CONCLUSIONS: The long term result of the TB control project in Jiangsu was good. The relapse rates in patients whose sputum converted negative within three months and beyond three months after the start of therapy were significantly higher than that in patients whose sputum converted negative within two months. PMID- 12783656 TI - [Evaluation of thrombolytic therapy by pulmonary radionuclide perfusion imaging in patients with acute pulmonary thromboembolism]. AB - OBJECTIVE: To assess the change of pulmonary perfusion after thrombolytic therapy in patients with acute pulmonary embolism. METHODS: Eighty patients with acute pulmonary embolism received pulmonary radionuclide perfusion imaging before and after thrombolytic therapy. Percentage of perfusion defect scores (PPDs) was calculated by semiquantitative analysis of pulmonary perfusion imaging before thrombolytic therapy (PPDsD(0)), 1 - 5 days (PPDsD(5)), 6 - 30 days (PPDsD(30)) and 3 months after thrombolytic therapy (PPDsM(3)). RESULTS: The mean PPDsD(0) of the 80 patients was (57.3 +/- 16.4)%. In 64 of the 80 patients, mean PPDsD(0) and PPDsD(5) were (55.5 +/- 16.8)% and (40.0 +/- 18.6)% respectively (P < 0.001). In 30 of these 64 patients, mean PPDsD(0), PPDsD(5) and PPDsD(30) were (57.5 +/- 16.1)%, (39.3 +/- 16.8)% and (29.5 +/- 17.3)% respectively. Differences between these 3 mean PPDs values were highly significant (all P < 0.001). In 11 of the 80 patients, mean PPDsD(30) and PPDsM(3) were (40.9 +/- 18.1)% and (29.1 +/- 27.1)% respectively (P < 0.05). In two groups of patients with the courses of disease in 7 days and in 8 - 14 days respectively, significant differences were found between PPDsD(0) and PPDsD(5) (P < 0.001, P < 0.001 respectively), and between PPDsD(0) and PPDsD(30) (P < 0.001, P < 0.005 respectively). The difference was also significant, although to a lesser degree (P < 0.05) between these values in a group of patients with the course of disease beyond 14 days. CONCLUSION: This study suggests that radionuclide pulmonary perfusion imaging is a reliable method for evaluating the changes of pulmonary perfusion before and after thrombolytic therapy in patients with acute pulmonary embolism. PMID- 12783657 TI - [Clinical-pathological study on 10 cases of pulmonary thromboembolism]. AB - OBJECTIVE: To improve the diagnosis of pulmonary thromboembolism (PTE) by a clinical-pathological analysis of 10 cases of PTE. METHODS: Eight cases of massive and submassive (in the segmental and larger pulmonary arteries) PTE confirmed by autopsy and 2 cases of PTE who underwent surgical treatment in the past 10 years (1991 - 2001) were studied. The specimens were fixed in 10% buffered formalin, and paraffin sections cut, and stained with HE and ET + VG. The pathological features were analyzed. RESULTS: There were 8 males and 2 females, aged 3.5 to 72 years (mean 33.3). The underlining diseases included congenital heart diseases (3 cases of ventricular septal defect complicated with pulmonary hypertension, and 1 case of tetralogy of Fallot), malignant lymphoma in the heart (n = 1), rheumatic heart disease (n = 1), lung metastatic cancer from stomach (n = 1), and phlebitis of the legs (n = 1). The sources of PE remained unknown in 2 cases. Among these cases, only 2 were diagnosed clinically. CONCLUSIONS: The research showed that PTE in patients who had underlining cardiovascular diseases, malignant tumors in the heart or lungs and pneumonia (pulmonary abscess) were hard to diagnose clinically. Pathological study is helpful in improving the clinical diagnosis of PTE. PMID- 12783658 TI - [Experimental study on lung carcinoma-targeted suicide gene therapy induced by irradiation]. AB - OBJECTIVE: To improve the efficacy and selectivity of gene therapy for lung carcinoma, a strategy was designed for suicide gene therapy in conjunction with irradiation therapy, by constructing a plasmid tgEgr-HyTK in which HSV-TK gene was driven by the early growth responsive gene-1 (Egr-1) promoter. METHODS: The radio-inducible suicide gene was constructed by insertion of Egr-1 promoter upstream of the HSV-TK gene. The expressions of HSV-TK in lung carcinoma cell lines A549 which were infected with tgEgr-HyTK and exposed to different doses of gamma-ray irradiation were analyzed, and the relative survival rates of cells in presence of prodrug ganciclovir (GCV) were tested. The tumor suppression effects were investigated in 40 nude mice bearing lung tumors to examine the efficacy of this Egr-TK gene therapy in vivo. RESULTS: Expression of HSV-TK gene in lung carcinoma cells infected with tgEgr-HyTK plasmids was markedly increased in a radiation dose-dependent manner. A gene therapy experiment in vitro showed that tgEgr-HyTK transduced lung carcinoma cells became highly sensitive to GCV after irradiation, but not without irradiation. tgEgr-HyTK transfected tumors regressed significantly after a combination therapy of irradiation and GCV in all mice (n = 10), and five tumors disappeared in 3 weeks without any side effect. CONCLUSION: The data indicate that tumor targeted expression of HSV-TK gene under the control of a radio-inducible promoter represents a novel strategy for safe and effective gene therapy of lung carcinoma, which may have clinical application in the future. PMID- 12783659 TI - [The TNF-alpha-induced apoptosis of LA795 cells is mediated by MAP kinase kinase 6]. AB - OBJECTIVE: To study the molecular mechanism of apoptosis of lung cancer cells stimulated by tumor necrosis factor alpha for the discovery of potential methods of gene therapy of lung cancer. METHODS: The apoptosis model of LA795 adenocarcinoma cells was performed. Recombinant adenovirus of MAP kinase kinase 6, and its constitutively active form and dominant negative form were prepared in 293 package cells. Protein kinase assay was used to detect the activity of MKK6 in cells. RESULTS: TNF-alpha stimulation increased the activity of MKK6 in LA795 lung cancer cells. The stimulation of TNF-alpha induced apoptosis of LA795 cells significantly. The infection of recombinant virus of constitutive active form of MKK6 also induced apoptosis of LA795 cells significantly, while the infection of recombinant virus of dominant negative form of MKK6 blocked the apoptosis of LA795 cells apparently. CONCLUSIONS: TNF-alpha induced apoptosis of LA795 cells is mediated by MKK6. It is possible to use MKK6 recombinant virus for gene therapy of lung cancer. PMID- 12783660 TI - [The effects of long-term application of L-arginine on K+ channel in pulmonary artery smooth muscle cells from rats exposed to chronic hypoxia]. AB - OBJECTIVE: To investigate the effects of long-term application of L-arginine (L Arg) on K(+) channel in pulmonary artery smooth muscle cells (PASMC) from rats exposed to chronic hypoxia. METHODS: Male Wistar rats were randomly divided into three groups: group A (control group), group B (chronic hypoxia group) and group C (chronic hypoxia plus L-Arg treatment group). The rats were exsanguinated and the intrapulmonary arteries (300 approximately 700 OD) were collected. Single PASMC was obtained by the acute enzyme separation method (collagnase Iplus papain) and the conventional whole-cell patch clamp technique was used to record resting membrane potential (Em), potassium ion currents of voltage-gated potassium channel and Ca(2+)-sensitive potassium channel. RESULTS: (1) The Em of group B [(-31 +/- 8) mV, n = 6] was significantly higher than that of group A [( 42 +/- 5) mV, n = 6, P < 0.01]. After long-term application of L-Arg, the Em of group C [(-39 +/- 4) mV, n = 6] was significantly lower than that of group B (P < 0.05). (2) The peak current at +50 mV of voltage-gated potassium channel was compared among three groups. In group B [(62.2 +/- 5.3) pA/pF, n = 6], it was significantly lower than that of group A [(121 +/- 9) pA/pF, n = 6] (P < 0.01). After long-term application of L-Arg, the peak current at +50 mV of voltage-gated potassium channel in group C [(95 +/- 3) pA/pF, n = 6] was significantly higher than that of group B (P < 0.001). (3) The peak current at +50 mV of Ca(2+) sensitive potassium channel was compared among three groups. In group B [(74.7 +/ 4.1) pA/pF, n = 6], it was significantly higher than that of group A [(53.6 +/- 5.9) pA/pF, n = 6] (P < 0.05). After long-term application of L-Arg, the peak current at +50 mV of Ca(2+)-sensitive potassium channel in group C [(31.8 +/- 1.8) pA/pF, n = 6] was significantly lower than that of group B [(74.7 +/- 4.1) pA/pF, n = 6] (P < 0.001). CONCLUSIONS: The long-term application of L-Arg can improve the resting membrane potential of PASMCs from rats exposed to chronic hypoxia, increase the currents of voltage-gated potassium channel and decrease the currents of Ca(2+)-sensitive potassium channel. These effects are considered to be effective in improving chronic hypoxic pulmonary hypertension. PMID- 12783662 TI - [Optimal margin in nephron-sparing surgery for renal cell carcinoma 4 cm or less in diameter]. AB - OBJECTIVE: To investigate the optimal margin in nephron-sparing surgery (NSS) for renal cell carcinoma (RCC) 4 cm or less in diameter. METHODS: Eighty-two kidneys with RCC 4 cm or less in diameter resected by radical nephrectomy were prospectively studied. The kidney samples were sectioned at 3 mm interval and examined for multicentricity. On each layer of tissue sectioned, parenchyma margin of 15 mm beyond pseudocapsule was continuously sectioned and examined for completeness of pseudocapsule and extra-pseudocapsule cancer lesion. The farthest distance between extra-pseudocapsule lesion and primary tumor was measured. PCNA expression was detected in 41 patients by using standard SP immunohistochemistry technique. RESULTS: The diameter of 82 primary tumors was 3.4 +/- 0.8 cm (range 1.5 - 4.0 cm). Of these, 31.7% (26/82) were found without intact pseudocapsule and 17.1% (14/82) with positive cancer lesions beyond pseudocapsule. The average distance between extra-pseudocapsule cancer lesion and primary tumor was 0.5 +/- 1.3 mm (range 0 - 5.0 mm), with a confidential interval (CI) of 95% (0.11, 0.94). Statistically, the one side percentile P(95) was 4.9 mm, P(97.5) was 5.0 mm and P(100) was 5.0 mm. The mean PCNA index in the 41 patients with RCC was (29.5 +/- 17.6)%, which was (49.6 +/- 21.5)% in the group with extra-pseudocapsule cancer lesions and (24.6 +/- 12.7)% in the group without (t = 3.162, P = 0.013). The ratio of strong expression was 5/8 in the group with extra-pseudocapsule cancer lesions, and 18.2% (6/33) in the group without the lesions (chi(2) = 6.442, P = 0.011). Logistic regression analysis showed that completeness of pseudocapsule and PCNA index were significant predictors of extra-pseudocapsule cancer lesions (P = 0.019). CONCLUSIONS: These data suggest that when NSS is performed in RCC 4 cm or less in diameter, a margin of more than 5 mm of adjacent parenchyma should be excised with the tumor. Enucleation alone was associated with a significant risk of incomplete excision, and therefore liable for local recurrence. Tumors with incomplete pseudocapsule and(or) high PCNA indices are more likely to have extra-pseudocapsule cancer lesions, so intensive follow-up is necessary after NSS. PMID- 12783661 TI - [Effect of levcromakalim and cromakalim on ATP-sensitive K+ channel of pulmonary arterial smooth muscle cells in pulmonary hypertensive rats]. AB - OBJECTIVE: To investigate the effect of levcromakalim and cromakalim on ATP sensitive K(+) channel (K(ATP)) and mean pulmonary arterial pressure (mPAP) in rats with chronic hypoxic pulmonary hypertension. METHODS: Ninety Wistar rats were divided randomly into a normal group (15) and a chronic hypoxia group (75). Rats in the hypoxia group were exposed to hypoxia (FiO(2) = 0.10 +/- 0.05) for 3 weeks, whereas rats in the normal group maintained in air. The currents of K(ATP) of pulmonary artery smooth muscle cells (PASMCs) from both the chronic hypoxic and the normal groups were observed using patch-clamp technique inside-out configuration, and the effect of levcromakalim and cromakalim on K(ATP) from chronic hypoxic group was determined under normoxic conditions. The mPAP and mSAP were determined by catheterization in hypoxic rats before and after levcromakalim and cromakalim were administrated respectively to the rats with doses ranging from 100 micro g/kg to 200 micro g/kg intravenously. RESULTS: Chronic hypoxia had little effect on the activity of K(ATP) in the chronic hypoxic rats. Levcromakalim and cromakalim remarkably activated the K(ATP) in chronic hypoxic rats. A dose-dependent decreasing effect on mPAP was elicited by levcromakalim and cromakalim in the hypoxic rats. CONCLUSION: Although K(ATP) activity is probably not directly involved in the development of chronic hypoxic pulmonary hypertension in rats, both levcromakalim and cromakalim decrease the increased mPAP induced by chronic hypoxia by activating K(ATP) of SMCs from pulmonary arteries, therefore diminishing the inhibitory effect of hypoxia on other K(+) channels. PMID- 12783663 TI - [Non-improvement after different procedures of prostatectomy]. AB - OBJECTIVE: To analyze the causes of non-improvement after different procedures of prostatectomy. METHODS: Using the urodynamics and other techniques, we examined 84 non-improved patients who had undergone different procedures of prostatectomy. RESULTS: In the TURP group, the main causes of non-improvement included bladder outlet obstruction (84.9%), weak contractility of the detrusor (30.4%), and detrusor instability (DI) (18.2%). In the open group, however, the main causes of non-improvement included detrusor weak contractility (52.9%), DI (35.2%), and bladder outlet obstruction (33.3%). There was significant difference between the two groups according to the Pearson Chi-square test (P = 0.000). The re obstruction rate in the TURP group was much higher than that in the open group, suggesting that the level of the TURP should be improved. CONCLUSIONS: Different operative procedures yield different non-improvement after prostatectomy gradually. When BPH patients are diagnosed with detrusor weak contractility and DI, prostatectomy should be carefully considered. PMID- 12783664 TI - [Applied anatomy of the path of retroperitoneal laparoscopy surgery]. AB - OBJECTIVE: To discover the anatomical basis of retroperitoneal laparoscopic surgery. METHODS: Twenty Chinese adult cadavers including 12 and 8 woman were anatomized. The relationship between nerves, vessels of the lateral abdominal wall, retroperitoneal structures and the path of retroperitoneal laparoscopic surgery were measured. RESULTS: In male cadavers, the distance between the iliac crest and lumbar trigonum was 4.02 +/- 1.26 cm on the left, and 3.83 +/- 0.90 cm an the right. The distance between the iliac crest and ilio-hypogastric nerve was 0.82 +/- 0.13 cm on the left and 0.84 +/- 0.08 cm on the right. The distance between the subcostal nerve and XII rib in the posterior axillary line and midaxilla line was 0.79 +/- 0.26 cm, 1.65 +/- 1.12 cm on the left and 0.78 +/- 0.30 cm, 1.59 +/- 1.07 cm on the right respectively. The distance between the ureter of inferior pole of kidney and extra-border of psoas was 2.24 +/- 0.67 cm on the left and 2.19 +/- 0.73 cm on the right. The distance between crossing of the ureter and iliac vessels and extra-border of psoas was 3.91 +/- 0.59 cm on the left and 3.76 +/- 0.53 cm on the right. In female cadavers, the distance between the iliac crest and trigonum lumbale was 3.90 +/- 0.37 cm on the left and 3.87 +/- 0.37 cm on the right. The distance between the iliac crest and ilio hypogastric nerve was 0.94 +/- 0.06 cm on the left and 0.93 +/- 0.10 cm on the right. The distance between the subcostal nerve and XII rib in the posterior axillary line and midaxilla line was 0.61 +/- 0.14 cm and 1.37 +/- 0.89 cm on the left and 0.64 +/- 0.13 cm, 1.32 +/- 0.95 cm on the right respectively. The distance between the ureter of inferior pole of kidney and extra-border of psoas was 2.00 +/- 0.23 cm on the left and 1.91 +/- 0.13 cm on the right. The distance between crossing of the ureter and iliac vessels and extra-border of psoas was 3.43 +/- 0.31 cm on the left and 3.85 +/- 0.43 cm on the right. CONCLUSIONS: The top of the lumbar trigonum which is 2 cm above the iliac crest selected as the first point and 2 cm inferior to the XII rib in the posterior axillary line selected as the second point is the anatomical basis for retroperitoneal laparoscopic surgery. It can reduce the opportunity to injury nerves, vessels and retroperitoneal structures while making retroperitoneal laparoscopic surgery easy. PMID- 12783665 TI - [Construction and screening of suppression subtractive hybridization library of renal cell carcinoma]. AB - OBJECTIVE: To construct and screen the suppression subtractive hybridization (SSH) library of human renal cell carcinoma (RCC). METHODS: Poly A(+) RNA was isolated from RCC lines 786-O (tester) and renal cell (RC) lines HK-2 (driver), respectively. SSH procedure was performed according to the protocol of the PCR Select cDNA Subtraction Kit (Clontech), and PCR products were cloned into pT-Adv vector and transformed E. coli TOP10F'. All positive clones picked out were digested and some of which were sequenced. RESULTS: The SSH library contained 362 clones with SSH cDNA fragments distributed mainly from 0.3 to 0.9 kb. Among 50 clones sequenced randomly, 2 represented unknown genes and the other 48 derived from 36 known genes. CONCLUSION: The quality of the SSH library of human RCC is reliable and its construction is the basis for further screening differentially expressed genes of RCC. PMID- 12783666 TI - [Expression and significance of clusterin in normal prostate, benign prostate hyperplasia and prostate cancer]. AB - OBJECTIVE: To investigate the expression and significance of Clusterin in normal prostate, benign prostate hyperplasia (BPH) and prostate cancer. METHODS: Clusterin expression in samples of 12 normal prostate, 15 BPH, and 56 prostate cancer were studied by immunohistochemical stain. RESULTS: Of 83 cases, 67 are positive or weak positive (81%). The rate of positive or weak positive for normal prostate, BPH and prostate cancer was 17% (2/12), 73% (11/15), and 96% (54/56) respectively. The expression level of Clusterin in prostate cancer was much higher than in normal prostate (t = 8.82, P < 0.01). BPH (t = 7.63, P < 0.01) was related positively with pathological grade (r = 0.649, P < 0.01) and stage (r = 0.609, P < 0.01) of prostate cancer. CONCLUSION: Clusterin may play an important role in the biological characteristics of prostate cancer by the anti-apoptosis pathway. PMID- 12783667 TI - [In vitro and in vivo functional evaluation of anti-human bladder tumor human mouse chimeric antibody ch-BDI]. AB - OBJECTIVE: To evaluate the in vitro and in vivo function of anti-human bladder tumor human-mouse chimeric antibody ch-BDI and its future clinical application. METHODS: With ch-BDI in high-expression cell-line medium, affinity chromatography was used for the purification. Labeled with (99m)Tc through reduction method, its immunoreactive fraction and association constant were measured. The constant was injected into nude mice with xenografted human bladder tumor. The biodistribution of the labeled ch-BDI was studied with radioimmunoimaging. RESULTS: ch-BDI showed desirable immunoreactive fraction (76%) and association constant (3.56 x 10(9) M( 1)) in vitro and a terrific specific targeting effect in vivo. CONCLUSION: ch-BDI has fairly good function against human bladder tumor both in vitro and in vivo, and is promising in clinical use. PMID- 12783668 TI - [Diagnosis and treatment of spinal dural arteriovenous fistulas: 110 cases report]. AB - OBJECTIVE: To discuss the diagnosis, treatment and prognosis of spinal dural arteriovenous fistulas (SDAVF). METHODS: The clinical and following-up date from 110 patients with SDAVF diagnosed by spinal MRI and spinal angiography were analyzed retrospectively. RESULTS: Draining vein between fistula and spinal cord was interrupted by laminectomy approach as first choice in 61 patients, by hemi laminectomy approach in 37, and by endovascular embolization in 12, and re operation due to recurrence after embolization in 3. Anticoagulation, hydration and early rehabilitation were used postoperatively. Complete disappearance of SDAVF was confirmed in all 106 patients who received postoperative spinal angiography. The para-medullary tortuous flow voids fully disappeared in 74 patients and partly disappeared in 15 on postoperative T(2)-weighted MRI. Follow up of 98 patients showed complete recovery in 54 patients, improvement in 34, and no change in 10. Three of the 10 patients were reoperated on because of recurrence 1 to 5 years after embolization. CONCLUSIONS: SDAVF can produce good outcome after early diagnosis and treatment. Interruption of the draining vein between fistula and spinal cord by hemi-laminectomy approach is the first choice for the treatment of SDAVF. PMID- 12783669 TI - [Diagnosis and treatment of cavernous sinus neurinoma]. AB - OBJECTIVE: To study the diagnosis and microneurosurgical treatment of cavernous sinus neurinoma. METHODS: Twenty-two patients with cavernous sinus neurinoma which had been diagnosed by MRI were operated on by microsurgery. Fourteen patients received excision of tumor through frontotemporal approach with zygomatic osteotomy and, 8 patients via subtemporal and suboccipital transtentorial approaches. RESULTS: Total resection was achieved in 21 patients (95.5%), and subtotal resection in 1. There was no operative death. The all operative samples were confirmed neurinoma pathologically. The symptoms postoperation such as headache and exophthalmos were reduced. Three months after operation, the function of the 6th nerve was improved in 12 of 19 patients with paresis of the 6th nerve. One patient with hemiplegia recovered well. The function of the 3th nerve recovered in 4 of 6 patients with recurrent paresis of the 3th nerve. Those patients with hydrocephalus were improved after operation. The patients were follow up 8 - 60 months (mean 20 months). No tumor recurrence was observed. CONCLUSION: Cavernous sinus neurinoma can be removed successfully via subtemporal and suboccipital transtentorial approaches. PMID- 12783670 TI - [Stereotactic microelectrode-guided posteroventral pallidotomy for Parkinson's disease]. AB - OBJECTIVE: To assess postoperative effects of microelectrode-guided posteroventral pallidotomy (PVP) for Parkinson's disease. METHODS: Intraoperative microelectrode recordings and microstimulation were used to explore the globus pallidus to performance of posteroventral pallidotomy in 48 patients with Parkinson's disease (47 unilateral and 1 bilateral). Assessment was made at baseline preoperatively and at 6 months intervals postoperatively, with unified Parkinson's disease rating scale (UPDRS). RESULTS: All patients were significantly improved on the limbs contralateral to the lesion side 6 - 34 months after operation (mean 24 months). The improvement was seen in the 'on' or 'off' state: UPDRS scores with patients on levodopa were improved by an average of 28.7%, while off medication scores showed reductions (47.6%) at 24 months. There were no deaths and no visual complications, but there were 4 patients (8.3%) of a delayed contralateral limbs dystonia after pallidotomy. CONCLUSIONS: The techniques of microelectrode recording and microstimulation indicate the location of the internal capsule and optic tract, which allow easy identification of these structure and facilitate PVP target in conjunction with radiofrequency microelectrode stimulation. PMID- 12783671 TI - [Clinical application of retrograde cerebral perfusion for brain protection during the surgery of ascending aortic aneurysm: 50 cases report]. AB - OBJECTIVE: To assess retrospectively the effects of different protective methods on brain in ascending aortic aneurysm surgery. METHODS: In 65 patients, aneurysm was dissected to the aortic arch or right arch. To protect brain, deep hypothermic circulatory arrest (DHCA) combined with retrograde cerebral perfusion (RCP) through the superior vena cava (n = 50) and simple DHCA (n = 15) were used during the procedure. Blood samples for lactic acid level from the jugular vein were compared in both groups at different phase, and perfusion blood distribution and oxygen content difference between the perfused and returned blood were measured in some RCP patients. RESULTS: The DHCA time was 35.9 +/- 18.8 min (10.0 - 63.0 min) and DHCA + RCP time was 45.5 +/- 17.2 min (16.0 - 81.0 min). The resuscitation time was 7.1 +/- 1.6 h (4.4 - 9.4 h) in DHCA patients and 5.4 +/- 2.2 h (2.0 - 9.0 h) in RCP patients. Operation death was 3/15 in the DHCA group and 1/50 in the RCP patients. Central nervous complication existed in 3/12 of DHCA patients and 1/49 of RCP patients (P < 0.01). The overall survival rate was 96% (RCP) vs 67% (DHCA), central nervous system dysfunction was 20% in DHCA vs 2% in RCP (P < 0.01). The blood lactic acid level increased significantly after reperfusion in DHCA than in RCP. The blood distribution measurement approximated to 20% of the perfused blood returned from arch vessels. Oxygen content between perfused and returned blood showed that oxygen uptake was adequate in the RCP group. CONCLUSIONS: The application of RCP could prolong the safety duration of circulation arrest. Cerebral perfusion may reep the brain cool and flush out particulate and air embolism. Open anastomosis of the aortic arch to the prosthesis can be safely performed. RCP is acceptable for brain protection in clinical practice. PMID- 12783672 TI - [Clinicopathological features of typical and non-typical hereditary non-polyposis colorectal cancer and their germline mutation of hMLH1 and hMSH2]. AB - OBJECTIVE: To study the clinicopathological features of the Chinese hereditary non-polyposis colorectal cancer and its germline mutation of hMLH(1) and hMSH(2). METHODS: Thirteen typical Chinese hereditary non-polyposis colorectal carcinoma (HNPC)C kindreds and 19 non-typical HNPCC families were registered and followed up. The germline mutation of the hMLH(1) and hMSH(2) of 12 index cases of 6 typical and 6 non-typical HNPCC were screened by PCR-SSCP. Samples with abnormal mobility were sequenced directly. RESULTS: The average age of typical HNPCC was 47, no difference existed between sexes. Location of the tumors of typical HNPCC represented 44.7% on the right half colon and non-typical HNPCC 65.8% on the rectum. The rate of the metachronos cancer was 11.5%. The 3-, 5-and 10-year survival rate was 64.0%, 45.3% and 31.2% respectively. Among 12 cases, 8 showed abnormal mobility. Except for an intron polymorphism, six exons abnormalities were found in 5 of 12 proband. Sequencing showed 4 missense, 7 insertion and a nonsense mutations. CONCLUSIONS: Chinese HNPCC is early onset, more common on proximal colon and better prognosis. Mutation of hMSH(2) is dominant in the Chinese typical HNPCC, but mutation of hMLH(1) is more common in the non-typical group. PMID- 12783673 TI - [Clinical analysis of 251 cases of soft tissue sarcomas]. AB - OBJECTIVE: To explore the therapeutic principles and prognostic factors of soft tissue sarcoma. METHODS: Two hundred and fifty-one patients with soft tissue sarcoma (STS) treated at Shanghai Cancer Hospital during 1986 - 1990 were reviewed retrospectively. RESULTS: The 1-, 3-, 5-, 10-year tumor-free survival rates were 67.74%, 57.16%, 52.41%, 38.60%, respectively. The overall survival rates for 1, 3, 5 and 10 years were 81.01%, 67.75%, 60.79%, and 49.23% respectively. Log-rank test showed that the patients with different pathological findings, histological grades, mass location and size, anatomical depth, and surgical margin showed different outcomes. Whether the sarcomas invaded the vessels or metastasized would influence the survival rates. The patients who underwent different interventions or operations also had different outcomes. The prognosis of STS was associated with age, histological type, histological grade, tumor size, surgical margin and metastasis according to the Cox regression analysis. CONCLUSION: During the treatment of STS, wide-resection, especially 3 dimensional resection, comprehensive treatment and individualized treatment should be advocated. PMID- 12783674 TI - [Expression of vascular endothelial cell growth factor and its receptor mRNA in breast cancer tissues]. AB - OBJECTIVE: To study the expression of vascular endothelial cell growth factor (VEGF) and its receptor FLT-1, FLK-1 mRNA in breast cancer tissues and their correlation with clinicopathological factors. METHODS: The expression of VEGF and and its receptor FLT-1, FLK-1 mRNA were analyzed by reverse transcription - polymerase chain reaction (RT-PCR) in the specimens from 47 patients with breast cancer and 11 patients with benign breast disease. RESULTS: VEGF121, 165 mRNA were all detected in malignant and benign breast tissues, with higher level, (0.420 +/- 0.133 and 0.291 +/- 0.094 respectively) in breast cancer tissues than in benign breast tissues, [0.196 +/- 0.067 (P = 0.000) and 0.206 +/- 0.058 (P = 0.001) respectively]. VEGF121 mRNA expression was stronger than VEGF165 mRNA expression (P = 0.000) in breast cancer tissues, whereas there is no significant difference in benign breast tissues (P = 0.666). FLT-1, FLK-1 mRNA were expressed in 18 of 47 (38.3%) and 12 of 47 (25.5%) breast cancer tissues respectively, but not in benign breast tissues. Moreover, no correlation was observed between the expression of VEGF121, VEGF165, FLT-1, FLK-1 mRNA in breast cancer tissues and patients' age, tumor size, lymph node metastasis, tumor stages, estrogen or progesterone receptor status. CONCLUSION: The expression of VEGF and its receptor FLT-1, FLK-1 mRNA were up-regulated in breast cancer tissues, suggesting its important role in angiogenesis of breast cancer. PMID- 12783675 TI - [Iodine-125 interstitial brachytherapy for malignant tumor]. AB - OBJECTIVE: To assess the clinical effects of (125)I interstitial brachytherapy for malignant tumors. METHODS: One hundred and twelve patients with malignant tumors of stage II stage and III under went radical resection combined with (125)I intraoperative implantation. Seven days and three month after operation WBC count and immune markers were observed. Blood biochemistry ultrasonography and X-ray were performed to observe recurrence and metastasis of tumors per three months. RESULTS: In the 112 patients, 3 died of tumor recurrence in six months, and others survived with the longest time of 35 months. CONCLUSION: (125)I interstitial brachytherapy for malignant tumor is simple, safe and effective. PMID- 12783676 TI - [Vascularized iliac bone grafting for avascular necrosis of the femoral head]. AB - OBJECTIVE: To evaluate the indications for vascularized iliac bone grafting in the treatment of osteonecrosis of the femoral head and operative results. METHODS: From October 1993 to August 2000, Twenty-six hips of 18 patients with osteonecrosis of the femoral head underwent vascularized iliac bone grafting. Clinical evaluation was made according to Harris Hip Score and ARCO staging system. RESULTS: Twenty-three hips of 16 patients were followed up for 31.5 months (6 - 74 months) on average. The mean Harris hip score was 76 patients (mean 61.7). Thirteen hips (medial 8 hips, central 3 hips, lateral 2 hips) were graded > 80 points by Harris hip score were. The incidence of radiographic collapse 20% was (medial), 40% (central) and 75% (lateral) respectively. CONCLUSIONS: Vascularized iliac bone grafting is effective for the treatment of osteonecrosis of the femoral head in short and middle term. We recommend this procedure for the patients with ONFH in ARCO stage I, stage IIA, IIB medial and central, and stage IIC medial. PMID- 12783677 TI - [Operative treatment of delayed acetabular fractures]. AB - OBJECTIVE: To assess the operative methods of delayed acetabular fractures and the operative results. METHODS: The operative approaches, procedures, results and complications of the delayed acetabular fractures between August 1993 and August 2001 in Jishuitan Hospital were evaluated retrospectively. RESULTS: Thirty-two patients were followed up 49.6 months on average. Sciatic nerve palsy was found in 1 patient. Excellent functional results of hip joints were found in 3 patients, good in 16, fair in 10, and poor in 3. Ectopic bone formation was observed in 6 patients and necrosis of the femoral head in 3. CONCLUSION: Careful selection of operative indications of delayed acetabular fractures in combination with proper operative approach and appropriate reduction and fixation, good results can be obtained. PMID- 12783678 TI - [Surgical management of metastatic disease of long bone]. AB - OBJECTIVE: To improve the life quality of cancer patients with metastasis to long bone and to select suitable surgical treatment. METHODS: Fifty two patients with metastasis 27 men and 25 women, were treated from 1990 to 1999. Their average age was 56.8 years (33 - 74). In 16 patients with multiple lesions, underwent surgery at bone shaft (29 patients) and bone epiphysis (26). Thirty patients were treated for pathologic fracture and the rest for impending fracture. Operations included limb-salvage (51 patients) and amputation (4) Limb salvage consisted of intralesional curettage (3 patients), intramedullary nailing reconstruction (29), endoprosthesis (18), and temporary spacer (1). 21 patients accepted postoperative chemotherapy or radiotherapy. RESULTS: Follow-up of 52 patients for a mean of 28.2 months (2 - 122 months) showed pain relief (41 patients), (75%) and full or part weight-bearing stability (36) 69%. Local tumor recurrence occurred in 11 patients. CONCLUSIONS: Surgical treatment can effectively improve the life quality of patients with metastasis to long bone. The metastatic lesions should be resected with wide or radical margin for the patients with kidney, breast, prostate and thyroid cancer. PMID- 12783679 TI - [Expression of fractalkine and its receptor in acute cardiac allografts rejection]. AB - OBJECTIVE: To investigate the expression of fractalkine (FKN) and its receptor CX3CR1 in cardiac allografts and the effect of Cyclosporin A (CsA). METHODS: Three groups of rats underwent heterotopic cardiac transplantation, 45 cases in each group and 5 cases in control group: SD to SD regarded as isograft group (group A), Wistar to SD divided into CsA untreated allograft group (group B) and CsA treated allograft group (group C), normal SD rats as control group. The FKN mRNA expression was detected by one-step RT-PCR method and the expression of FKN and CX3CR1 protein was detected by standard ABC immunohistochemical technique. RESULTS: The expression of FKN mRNA and protein was weak in both isografts and normal heart specimens. The changes of FKN mRNA expression were correlated with the process of acute allograft rejection. The peak of FKN mRNA expression (0.8 +/ 0.26) appeared on the seventh day after transplantation, which could be down regulated by CsA significantly (t = 2.390, P < 0.05). FKN protein was located in endothelia cells and its receptor CX3CR1 was located in infiltrating mononuclear cells in allografts. CONCLUSIONS: Upregulation of FKN and its receptor was significantly correlated with the trafficking of mononuclear cells which play an important role in acute allograft rejection. It may be one of the important mechanisms of CsA to intervene the acute rejection by inhibiting the activation of the FKN-CX3CR1 pathway. PMID- 12783680 TI - [Dual cell seeding to improve cell retention on polytetrafluoroethylene grafts]. AB - OBJECTIVE: To improve cell retention on the graft with dual cell seeding technique, adding a layer of smooth muscle cells (SMCs) between the graft and endothelial cells (ECs). METHODS: Polytetrafluoroethylene (PTFE) grafts pretreated with fibronectin were seeded with SMC followed by ECs one day later and exposure to an in vitro flow system. The number of ECs on grafts was counted under microscope. RESULTS: After exposure to the flow for 1 hour, 61% of the ECs lost when the ECs were seeded alone, whereas only 27% of the ECs lost when the ECs were seeded on the top of SMCs. Preconditioning of a cell seeded graft in a low rate flow did not improve cell retention when late exposure to a higher rate flow. CONCLUSION: This in vitro study indicates that using SMCs as a media layer between ECs and graft surface can improve the retention of seeded ECs on PTFE graft. PMID- 12783681 TI - [Effects of enteral nutrition on uptake of amino acid and enzyme-protein synthesis of pancreatic acinar cell in acute pancreatic dogs]. AB - OBJECTIVE: To evaluate the effect of intrajejunal nutrition on uptake of amino acid and enzyme-protein synthesis in pancreatic acinar cell and subcellular fractionation and zymogen granules in dogs with acute pancreatitis. METHODS: Fifteen dogs were induced acute pancreatitis by retrograde injection of 5% sodium taurocholate into the pancreatic duct. Radioactive tracing and electron microscope were used to evaluate the change of amino acid uptake, enzyme-protein synthesis in acinar cell, subcellular fractionation, the quantitative analysis of mean zymogen granule number and mean zymogen granule area after injection L-(3)H phenylalanine 30, 60, 120 1nd 180 min on the 7(th) day. RESULTS: The radioactivity of L-(3)H phenylalanine uptake by pancreatic acinar cells and incorporations of L-(3)H phenylalanine into newly synthesized enzyme-protein peaked at 60 min. In enteral nutrition (EN) group it was higher that that in parenteral nutrition (PN) group (P < 0.05), and then gradually declined. The radioactivity peaked at 60 min in zymogen granule, lysosomal-mitochondria and microsomal subcellular fractionation. The latter two decreased, bat there was no significant difference (P > 0.05). The change of the mean number and mean area of zymogen granules were not significant different between the EN group and PN group (P > 0.05). CONCLUSION: EN or PN do not stimulate pancreatic acinar uptake amino acid and enzyme-protein synthesis in acinar cell and subcellular fractionation. PMID- 12783682 TI - [Correlation of tumor angiogenesis with clinicopathologic prognostic parameters in choroidal melanoma]. AB - OBJECTIVE: Tumor angiogenesis is an essential step for tumor growth, progression and metastasis. Vascular endothelial growth factor (VEGF) is a mitogen specific for vascular endothelial cells, and plays a key role in tumor angiogenesis. The purpose of the present study was to investigate the expression of VEGF mRNA and VEGF protein in choroidal melanoma and to correlate these factors and the microvessel density (MVD) with the clinicopathologic prognostic features. METHODS: In a total of 65 cases of choroidal melanoma from the Pathology Laboratory of Tongren Hospital, the expression of VEGF mRNA was assessed by in situ hybridization (ISH); the expression of VEGF protein and factor VIII-related antigen (FVIII-RAg) was assessed using immunohistochemical methods. MVD was evaluated by immunohistochemical staining of FVIII-RAg for vascular endothelial cells. RESULTS: VEGF mRNA expression was present in 56/65 cases (86%), while VEGF immunoreactivity was present in 55/65 cases (85%). Positive expression of VEGF and VEGF mRNA was correlated with the tumor size (P = 0.025) and local invasion of tumor to scleral emissaria (P = 0.034), but it's relation to the histologic type, the tumor location and the largest tumor diameter (LTD) was not statistically significant. MVD was significantly higher in the VEGF-positive melanoma than in the VEGF-negative melanoma (one way analysis of variance, P = 0.000). There was no significant correlation between MVD and tumor location, as well as the tumor height and the patient's age; however, significant correlation was found between MVD and LTD, as well as with the tumor size, cell type and the invasion of tumor to the scleral emissaria (t test, P = 0.012, 0.008, 0.001 and 0.000, respectively). CONCLUSIONS: Most choroidal melanomas express VEGF mRNA and VEGF protein, and this expression is correlated with MVD. This suggests that angiogenesis is significantly correlated with clinicopathologic prognostic factors of choroidal melanoma, such as LTD, tumor size, cell type and tumor invasion. MVD and the expression of VEGF may be useful prognostic indicators for the biological behavior and the outcome of choroidal melanoma. PMID- 12783683 TI - [Effect of bcl-2 antisense oligonucleotides on multidrug resistance of cultured uveal melanoma cells]. AB - OBJECTIVE: To evaluate the chemotherapeutic sensitivity of uveal melanoma in vitro and reverse its drug resistance by the delivery of bcl-2 antisense oligodeoxynucleotide (AS-ODN). METHODS: The drug sensitivity tests of primary cultured uveal melanoma cells toward 5-flurouracil (5-FU), thiophosphamide (TSPA), cisplatin (DDP), adriamycin (ADM), vinblastine (VLB), dacarbazine (DTIC) were performed with 3, -4, 5 dimethyliazol-2,5 diphenyl tetrazolium bromide (MTT). AS-ODN bcl-2 were delivered with cationic lipid to down-regulate bcl-2 expression. Immunohistochemistry and Western-blot methods were used to detect bcl 2 expression. According to the principle of multi-drug mutual action, the influence of anti-apoptosis gene bcl-2 on tumor cell drug sensitivity was measured. RESULTS: Uveal melanoma was resistant to different chemotherapeutic drugs in different degrees. AS-ODN bcl-2 could down-regulate bcl-2 expression, was synergetic with all tested drugs and partially reversed the multi-drug resistance. CONCLUSIONS: Clinically, with ordinary dosages the above 6 drugs have little cytotoxicity to uveal melanoma cells, bcl-2 over-expression is associated with multi-drug resistance and AS-ODN bcl-2 can partially reverse the multi-drug resistance. PMID- 12783684 TI - [Clinical analysis of tumors of the iris and ciliary body]. AB - OBJECTIVE: To evaluate the clinical features, different diagnosis, pathological features, management, and prognosis of tumors of the iris and ciliary body. METHODS: Medical records, photographs, pathological findings and the results of follow-up of 37 cases with tumors of the iris and ciliary body were reviewed as a retrospective study. RESULTS: Of the 37 cases with tumors of the iris and ciliary body, 26 were male and 11 were female. The mean age at diagnosis was 38 years, ranged from 5 - 64 years. According to the histopathological examination, melanoma was the most common tumor in the iris and ciliary body (15 cases, 40.5%), followed by metastatic tumors (8 cases, 21.6%), teratoid medulloepitheliomas (3 cases, 8.1%) and iris nevus (2 cases, 5.4%). MANAGEMENT: The tumors were excised in 14 cases. Enucleation was performed in 21 cases. Two cases were observed without any surgical treatment. Thirty-four cases were followed-up for 2 months to 15 years, averaged 31 months. Most melanomas of the iris and ciliary body are round or semi-spherical dark brown vascularized mass, with engorged episcleral sentinel vessels in some cases. The tumor showed a shadow during transillumination. Ultrasound biomicroscopy revealed a low to medium echoic solid lesion, with echoic changes in adjacent infiltrated tissues. Melanoma showed positive immunoreactivity for melanoma-specific antigen, and had a good prognosis. Metastatic tumors of the iris and ciliary body were flat or near round, dirty, single or multiple neoplasms, growth rapidly, with abundant neovascularization, and had a poor prognosis. Primary carcinomas could be found in other parts of the body. CONCLUSIONS: Melanoma of the iris and ciliary body has typical features that may help to distinguish them from other tumors. Metastatic tumor has characteristic features, but the diagnosis can be made only with supplementary examination and immunocytochemical studies. Medulloepitheliomas should be differentiated from retinoblastoma. PMID- 12783685 TI - [Clinical and pathologic analysis of orbital pseudotumor]. AB - OBJECTIVE: To investigate the relationship between the histopathology and the imaging analysis of orbital pseudotumor to judge the histopathological types of these tumors before treatment according to imaging analysis, in order to search for different therapeutic approaches for different pathologic types and to predicate prognosis. METHODS: Fifty-six cases (58 eyes) of orbital pseudotumor that had been operated in Tongren hospital were studied. The clinical manifestations of these patients were reviewed. Changes in the imaging analysis and histopathologic examination were observed and their relationship was studied. Methods of treatment and their prognosis were also analyzed. RESULTS: Historically, the lymphocytic infiltrated type of orbital pseudotumor composed of large amounts of chronic inflammatory cells, only a small amount of collagen was found, so the imaging features of the lymphocytic infiltrated type presented as low or none reflecting areas with a clear posterior demarcation on B scan, and the signals were low or equal on T(1)WI and high on T(2)WI, which could be enhanced by contrast. Oppositely, the fibrotic type was composed of large amounts of fibrotic tissue, and few chronic inflammatory cells were found, so that only the anterior border of the lesion could be identified and the posterior demarcation was blur on the B scan, and the signals were low or equal on T(1)WI and low on T(2)WI, which could not be enhanced or could be enhanced only slightly by changing the contrast. The mixed type presented as an admixture of chronic inflammatory cells and fibric hyperplasia, and the imaging of the mixed type was midway between the lymphocytic infiltrated type and the fibrotic type. Forty-two patients of lymphocytic and mixed types which were identified by histopathology and imaging analysis received a local and systemic corticosteroid therapy. The effect was unstable, and recurrence occurred easily. Therefore, operations were performed on these patients, thirteen of them were treated with low-dose radiotherapy (2 000 - 3 000 cGy) two weeks after operation. Fifteen cases recurred after the operation and 4 cases recurred after the radiotherapy. The fibrotic type of orbital pseudotumor was identified in 12 cases in which no effect could be observed after conservative therapy and operations were performed subsequently. Two cases recurred and were operated again. Exenteration was performed in 2 cases of 56 cases. The cure rate of the composite treatment in the present series was 68.5%. CONCLUSIONS: Different types of orbital pseudotumor present different imaging exhibition, and imaging features are closely related with histopathological changes. Lesions can be localized by CT scan, and also can be localized by B scan and MRI. While it is difficult to obtain a cure result in the orbital pseudotumor, the cure rate can be improved by composite treatment. PMID- 12783686 TI - [The failure causes of non-penetrating trabecular surgery and reoperation]. AB - OBJECTIVE: To study the failure causes of non-penetrating trabecular surgery (NPTS) with SKGeL (a hyaluronic acid biological gel) implant and the surgical method of reoperation. METHODS: Repeated operation with mitomycin (MMC) through the initial surgical site was performed on 13 failure cases (13 eyes) that had undergone NPTS with SKGeL implant. The blockage of filtration tract was removed and the anterior chamber was intact during the surgery. All of these cases were open-angle glaucoma. Before the repeated surgery ultrasound biomicroscopic (UBM) examination was performed on the primary filtering bleb, and the intraocular pressure (IOP) examination was followed after the repeated operation. RESULTS: The mean follow-up period was (14.0 +/- 5.8) months (6 to 24 months). The examination of UBM showed that the filtering bleb disappeared and there was a liquid chamber under the superficial scleral flap in every failure case. The filtration failure due to the scarring at conjunctiva-Tenon's capsule-superficial scleral flap interface in 9 cases, proliferative membrane formation on the trabecular surface in 3 cases, micro-penetration of the trabecula in 1 case. At the end of follow-up, the IOP of 10 cases was lower than 21 mmHg without medication, the mean IOP level was (14.1 +/- 3.7) mm Hg, the IOP of 1 cases was 15 mmHg with Betagen, another 2 cases failed again 6 months after the repeated surgery and underwent the trabeculectomy at last. The complications included hyphema in one case and micro-penetration of the tabecula in one case. CONCLUSIONS: The scarring at conjunctiva-Tenon's capsule-superficial scleral flap interface was the most important cause of NPTS with SKGeL implant failure. A repeated surgery with MMC through the initial surgical site may be a choice for the failure cases. PMID- 12783688 TI - [Comparing patients' quality of life after phacoemulsification with intraocular lens implantation with that after extracapsular cataract extraction with intraocular lens implantation]. AB - OBJECTIVE: To Compare the effects of phacoemulsification with intraocular lens implantation (PHACO + IOL) to extracapsular cataract extraction with intraocular lens implantation (ECCE + IOL) on quality of life. METHODS: The study population consisted of 116 patients receiving PHACO + IOL and 93 patients receiving ECCE + IOL. They were interviewed using the quality of life questionnaire, and the clinical outcomes were obtained before surgery and in 1 week, 1 month, 3 months after surgery respectively. RESULTS: Patients receiving PHACO + IOL reported larger benefit in quality of life and all sub-scales than did those receiving ECCE + IOL in 1 week after surgery. Subjects underwent PHACO + IOL showed better improvement in quality of life and two sub-scales (social and mental) than did those underwent ECCE + IOL in 1 month and 3 months after surgery. However, the improvement in other two sub-scales (self-care and mobility) was similar between two surgical groups in 1 month and 3 months after surgery. CONCLUSION: The patients receiving PHACO + IOL reported better and more rapid improvement in quality of life within 3 months after surgery. PHACO + IOL and ECCE + IOL have the same effects on improvement in self-care and mobility. So quality of life in patients receiving PHACO + IOL are better than those of ECCE + IOL. PMID- 12783687 TI - [Phacoemulsification and intraocular lens implantation after scleral buckling surgery]. AB - OBJECTIVE: To determine the intraoperative and postoperative complications and best-corrected visual acuity outcomes of eyes undergoing phacoemulsification and intraocular lens (IOL) implantation after retinal detachment repair by scleral buckling technique. METHODS: The charts of all patients who underwent phacoemulsification and IOL implantation between May 1996 and July 2000 in Zheyi Eye Center were reviewed to identify the eyes with a history of retinal detachment repaired by scleral buckling technique. RESULTS: We identified 33 eyes of 33 patients. The mean interval from phacoemulsification to final examination was (1.9 +/- 1.0) years. The final best-corrected visual acuity of >or= 20/200 was attained in 28 (84.8%) of 33 eyes and >or= 20/50 in 12 (36.4%) of the eyes. With regard to complications, 2 eyes required laser capsulotomy for posterior capsular opacification. No eye developed a retinal re-detachment during 2 years follow-up. CONCLUSION: Phacoemulsification and IOL implantation can be performed safely after scleral buckling surgery and excellent best-corrected visual acuity results can be attained in most eyes. No retinal re-detachment occurred in this series. PMID- 12783690 TI - [Significance of systematic retroperitoneal lymphadenectomy at second-look laparotomy for ovarian cancer]. AB - OBJECTIVE: To investigate the timing and role of systemic retroperitoneal lymphadenectomy in patients with ovarian cancer. METHODS: From Jan. 1987 to Feb. 1994, 50 patients with ovarian cancer, who underwent retroperitoneal lymphadenectomy on second-look laparotomy (SLL), were retrospectively studied. RESULTS: The median age was 49 years. Overall survival at 3 and 5 years were 72% and 62%, respectively. Twenty of 50 (40%) women were found SLL(+), and the rates of positive SLL were related to International Federation of Gynecology Obstetrics (FIGO) stage, with 16% in stage I and II, 64% in stage III and IV (P < 0.01). Sixteen (32%) women were found to have retroperitoneal lymph nodes metastases at SLL, with 0% (0/15), 20% (2/10), 54% (13/24), 1/1 in stage I, II, III, IV respectively. In patients with SLL(+), 4 patients with sole disease in the pelvis, and 6 (12%) women only with micro-metastases of retroperitoneal lymph nodes. Fifteen patients with stage I ovarian cancer were all SLL(-), and SLL(-) were in 6 (60%) patients with stage II disease and 11 (38%) in stage III. There is no disease recurrence in patients with negative SLL till the data censored. CONCLUSION: Timing of systemic retroperitoneal lymphadenectomy at SLL is rational, and may aid in reducing disease recurrence of negative SLL and improving five-year survival rate in patients with ovarian cancer, especially in those with advanced epithelial ovarian cancer. PMID- 12783689 TI - [Inhibition of lens epithelial cell cycle and PCNA mRNA expression by an antisense oligonucleotides complementary to the messenger RNA of proliferating cell nuclear antigen]. AB - OBJECTIVE: To determine whether antisense oligonucleotides complementary to the messenger RNA of proliferating cell nuclear antigen (PCNA ASODN) inhibit the proliferation of bovine lens epithelial cell (BLEC) by changing the cell cycle and down-regulating the expression of PCNA mRNA and PCNA protein. METHODS: BLECs were cultured in vitro, and the second passage cells were used in this experiment. PCNA ASODN (30 micro mol/L), PCNA SODN (sense oligonucleotides, 30 micro mol/L), basic fibroblast growth factor (bFGF, 10 micro g/L), bFGF (10 ng/ml) + ASODN (30 micro mol/L), bFGF (10 micro g/L) + SODN (30 micro mol/L) were introduced respectively into the medium, and the same amount of PBS was added into the medium as a control. After 24 hours, the cell cycle and the PCNA expression were counted by flow cytometry, and the expression of PCNA mRNA was indicated by Northern ELISA hybridization. RESULTS: PCNA ASODN could decrease the rate of the cells of S phase and down-regulate the expression of PCNA mRNA and PCNA protein. Comparing with the control group, after 24 hours, the rate of cells of S phase was decreased from 15.67% to 7.96%, the expression of PCNA mRNA from 0.266 to 0.176 and the expression of PCNA protein from 55.27% to 12.32%. PCNA ASODN could also inhibit the proliferation of BLEC induced by bFGF, comparing with the bFGF group, the rate of cells of S phase was decreased from 23.4% to 19.9%, the expression of PCNA mRNA from 0.576 to 0.357 and the expression of PCNA protein from 76.4% to 35.48%. CONCLUSIONS: Our results demonstrate that the PCNA ASODN decreases expression of PCNA mRNA and PCNA protein, and stops the cell to enter and progress through S phase. These results provide an important impetus to initiate in vivo studies to determine the feasibility of antisense strategies in the prevention of posterior capsular opacification. PMID- 12783691 TI - [Study on serum vascular endothelial growth factor level in ovarian malignant tumors]. AB - OBJECTIVE: To explore the diagnostic value of serum vascular endothelial growth factor (VEGF) level in patients with ovarian carcinomas, and to evaluate the correlation between serum VEGF level and prognosis of ovarian cancer. METHOD: Serum VEGF level in patients with ovarian neoplasms (120 cases of ovarian carcinomas, 25 cases of benign tumors) and 90 healthy women as controls was determined by enzyme linked immunosorbent assay (ELISA), also the change of preoperative and postoperative serum VEGF level of 25 cases of epithelial ovarian cancer was observed. RESULTS: (1)The preoperative serum VEGF levels were (766 +/- 1237) mg/L in malignant groups, and there was a significant difference compared with normal controls (55 +/- 19) mg/L or patients with benign tumor (56 +/- 23) mg/L (P < 0.05). As a cut-off at 100 mg/L, sensitivity and specificity of preoperative serum VEGF levels for diagnosing ovarian cancer was 77% and 87% respectively. (2) Serum VEGF level were significantly elevated in advanced stage (955 +/- 1716) mg/L and poorly differentiated (991 +/- 1349) mg/L compared with early stage (198 +/- 287) mg/L and well differentiated (280 +/- 552) mg/L (P < 0.05), but there is no significant difference between the VEGF level and pathological types (P > 0.05). (3) The postoperative serum VEGF levels (118 +/- 110) mg/L were significantly lower than that in preoperative (1074 +/- 1211) mg/L. (4) The average total survival-time in patients with VEGF(+) was 28 months, but it in patients with VEGF(-) was 35 months (P < 0.05). CONCLUSION: Serum VEGF values were higher in malignant tumors and preoperative serum VEGF level has a potential as a marker for diagnosing and monitoring in ovarian neoplasms. PMID- 12783692 TI - [Effect of retroperitoneal lymphadenectomy on survival of patients with epithelial ovarian cancer]. AB - OBJECTIVE: The purpose of this study was to determine prognostic factors that have an impact on overall survival and to assess the rational application of retroperitoneal lymphadenectomy in patients with epithelial ovarian cancer. METHODS: A retrospective review was performed of 131 patients treated between Jan.1990 and Dec.1998 in Union Hospital and Tongji Hospital. Survival was calculated by Kaplan-Meier method and comparison was performed using Log-rank test. Independent prognostic factors were identified by the COX proportional hazards regression model. RESULTS: Multivariate analysis showed that the age, stage, residual tumor, retroperitoneal lymphadenectomy and the number of courses of chemotherapy were the most important prognostic factors. The overall 5-year survival was 66% and 41% for patients who did and did not undergo lymphadenectomy, respectively (P < 0.01). But the survival rate could not be improved through retroperitoneal lymphadenectomy in the patients with early stage, advanced stage whose residual tumor > 2 cm and those with mucinous adenocarcinoma (P > 0.05). Among patients with advanced stage whose residual tumor < or = 2 cm, 5-year survival was 65% and 30% for patients who did and did not undergo lymphadenectomy, respectively (P < 0.01). Among patients with serous adenocarcinoma, 5-year survival was 61% and 31% for patients who did and did not undergo lymphadenectomy, respectively (P < 0.01). CONCLUSIONS: The prognosis of the patients with epithelial ovarian cancer may be influenced by age, stage, residual tumor, retroperitoneal lymphadenectomy and the number of courses of chemotherapy. Although retroperitoneal lymphadenectomy could improve the survival rate, it should be carried out selectively. PMID- 12783693 TI - [Clinical and pathological features of borderline ovarian tumors]. AB - OBJECTIVE: To study the relationship between clinical pathological factors and prognosis. METHODS: Thirty-four cases with borderline ovarian tumors (BOT) and 30 cases with stage I ovarian epithelial cancer admitted in our hospital from Jan.1973 to Dec.2000 were studied retrospectively. All the cases were diagnosed according to the International Histological Classification and Staging of Ovarian Tumors (WHO, 1999). RESULTS: Thirty-seven cases were finally diagnosed of BOT, 6 cases with "microinvasive" had been misinterpreted as stage I ovarian cancer and one had "non-invasive peritoneal implants". Serous (38%) and mucinous (51%) tumors were dominant type of BOT and 95% of tumors were at stage I (International Federation of Gynecology and Obstetrics, FIGO). All patients were operated, 11 cases given conservative surgery with a recurrence rate of 9%, 26 cases had adjuvant chemotherapy, cyclophosphamide + adriamycin + cis-platinum used mostly. 5-year and 10 year survival rates were both 100%. The prognosis of BOT is related to pathology and adjuvant therapy. CONCLUSION: Surgery is the main treatment choice of BOT, and the criteria of chemotherapy must be adequately used. PMID- 12783694 TI - [Changes of relative variables about prethrombosis state and its clinical significance in patients with pregnancy induced hypertension]. AB - OBJECTIVE: To explore the changes of the hemorrheology coagulation and fibrinolytic system in patients with pregnancy induced hypertension (PIH). METHODS: To examine the platelet alphagranule membrane protein (GMP-140), Von Willebrand factor (vWF), antithrombin III activity (AT-III), plasminogen activity (PLG), the activity of tissue-type plasminogen activator (t-PA) and plasminogen activator inhibitor (PAI) in 30 non-pregnancy women, 30 normal late pregnancy women and 50 pregnancy induced hypertension women. RESULTS: In PIH group vWF [(156 +/- 23)%], GMP-140 [(23.9 +/- 3.6) microg/L], Fbg [(15.5 +/- 0.8) g/L], PLG [(141 +/- 9)%], D-Dimer [(1.42 +/- 0.32) mg/L], PAI [(9.2 +/- 0.8) kU/L] were obviously higher than those of control group. AT-III, ProC in PIH group were lower than that of the control group (P < 0.01). t-PA has no significant differences (P > 0.05). CONCLUSIONS: Prethrombotic state was shown in normal late pregnancy women. Obvious prethrombotic state was shown in patients with pregnancy induced hypertension women. PMID- 12783695 TI - [Study on regulatory effects of interleukin-6 on placental growth factor expression in trophoblast cells]. AB - OBJECTIVE: To investigate the expression pattern of Placental growth factor (PlGF) and regulatory effects of interleukin-6 (IL-6) on PlGF expression in the primary cultured trophoblast cells. METHODS: Cytotrophoblast cells were collected by trypsin-collagenase digestion and Percoll gradient centrifugation for primary culture. Reverse transcriptase polymerase chain reaction (RT-PCR) were resorted to demonstrate the expression pattern of PlGF mRNA in trophoblast cells cultured in vitro. The effects of IL-6 with different concentration (100, 10, 1 and 0.1 microg/L) and duration (6, 12, 24 and 48 h) on PlGF expression were observed. RESULTS: The 185 bp and 248 bp bands of PlGF were showed by RT-PCR. PlGF expression correlated with IL-6. PlGF began to increase at 6 h, and reached the climax at 12 h when recultured with 100 microg/L IL-6. CONCLUSIONS: PlGF expression has time and dose dependence on IL-6. It may play an important role in early pregnancy. PMID- 12783696 TI - [Effects of mifepristone on ultrastructure of placental Hofbauer cells in early human pregnancy]. AB - OBJECTIVE: To investigate the effects of mifepristone on the ultrastructure of Hofbauer cells in human early pregnant placenta. METHODS: Twenty 6 - 9 week pregnant women with indications of pregnancy termination were recruited and randomly assigned to mifepristone group(n = 10) and D & C group (n = 10). Villi were collected and studied with transmission electron microscope. RESULTS: In comparison with the control group, the ultrastructure of Hofbauer cells of mifepristone group showed the following changes: the cells were markedly edematous. The number of cytoplasmic processes of Hofbauer cells deceased obviously. In the cytoplasm of Hofbauer cells, the size of vacuoles enlarged and of mitochondria minimized. Rough endoplasmic reticulum and Golgi complex were under-developed. Lysosomes were rare. The nuclei enlarged and showed irregular shape. CONCLUSIONS: Mifepristone may change the phagocytosis' water and electrolyte transportation and immunological function of Hofbauer cells by influencing the ultrastructure of the Hofbauer cells. Therefore it can influence the development of pregnancy. PMID- 12783697 TI - [Association study between a polymorphism of aldosterone synthetase gene and the pathogenesis of polycystic ovary syndrome]. AB - OBJECTIVE: To investigate the relationship between -344T polymorphism in aldosterone synthetase (CYP11B2) gene promoter region and the pathogenesis of polycystic ovary syndrome (PCOS). METHODS: Ninety two patients with PCOS and controls were genotyped according to the fragment length (273 bp and or 202 bp) of CYP11B2 gene promoter by the technique of polymerase chain reaction restriction fragment length polymorphism. The levels of luteinizing hormone, follicular stimulating hormone, estrodiol, progesterone, prolactin, testosterone, plasma renin activity (PRA), plasma angiotensin II (PANG II) and aldosterone in the basal state were also determined. Different genotypes between PCOS were compared about their levels of PRA, PANG II, aldosterone and testosterone. RESULTS: (1) The C allele frequencies of CYP11B2 gene in control and PCOS was 22% and 36%, respectively. (2) The frequency of variants (TC, CC) of CYP11B2 gene 344T polymorphism site in PCOS (57%) was significantly higher than that of control subjects (37%). (3) The level of PRA, PANG II, aldosterone, testosterone were all significantly higher in the genotype of -344CC than in that of -344TT in PCOS and normal women (P < 0.01). CONCLUSIONS: (1) The variants (T-->C) of -344T polymorphism site of CYP11B2 gene predisposes increased risk of PCOS. (2) The genotype of -344CC, -344TC may be susceptible genotype of PCOS and has related to the enhanced functional activity of ovarian renin angiotensin system in PCOS. PMID- 12783698 TI - [Study on serum inhibin concentrations in women during menopausal transition]. AB - OBJECTIVE: To investigate the changes of serum inhibin (Inh)-A, Inh-B concentrations during menopausal transition and the time relationship between changes of serum Inh-A, Inh-B and other reproductive hormone levels. METHODS: Serum Inh-A, Inh-B concentrations were measured by Serotec modified two-site enzyme immunoassay during different phases of normal menstrual cycle in 10 healthy reproductive women, any time of 10 postmenopausal women. So did the serum Inh-A on 5 - 9 days prior to next period (premenstrual phase) and Inh-B determinations on the 3rd day of menstrual cycle in 40 women of age 43 - 52 (menopausal transition group). In addition, serum follicle stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E(2)), progesterone (P) levels were also determined when necessary for the purpose of analysis. RESULTS: The fluctuation patterns of serum Inh-A, Inh-B concentrations during normal menstrual cycle were completely different. About 48% of women during menopausal transition had normal luteal function as shown by the P levels during the premenstrual phase. The only significant change in these women as compared with the young was decrease of Inh-A concentration [(24.7 +/- 13.0) ng/L Vs (42.9 +/- 12.1) ng/L, P = 0.017] in the same phase. Further significant declines of serum Inh-A levels were seen in the luteal phase defect (LPD) and anovulatory (AOV) groups [(12.4 +/ 10.2) ng/L and (5.3 +/- 3.8) ng/L, P = 0.033, P < 0.000 1 respectively], until undetectable in the postmenopausal group. The day 3 Inh-B levels tended to decrease in the normal luteal and LPD groups, became undetectable in the AOV and postmenopausal groups (P = 0.001). Day 3 Inh-B levels was significantly lower in women with day 3 FSH > or = 10 IU/L than those with < 10 IU/L [(16.2 +/- 4.0) ng/L Vs (62.0 +/- 43.8) ng/L]. The elevation of day 3 FSH, LH levels was not significant until the AOV group (P = 0.009, P = 0.027 respectively), and the drop of E(2) levels until the postmenopausal group (P < 0.001). CONCLUSIONS: It is suggested that serum reproductive hormones should be measured in women of menopausal transition in order to know the stage of menopausal transition and to guide the clinical management. The decrease of serum Inh-A levels during the premenstrual phase is the earliest change of menopausal transition, and decrement of day 3 Inh-B levels a marker of decreased ovarian reserve. PMID- 12783699 TI - [Study of angiogenesis in human colorectal carcinoma and its modulation by p53 and K-ras gene]. AB - OBJECTIVE: To investigate the angiogenesis in human colorectal carcinoma and its modulation by p53 and K-ras gene. METHODS: The positive rate of p53 and K-ras gene mutation, the expression of vascular endothelial growth factor (VEGF) and microvessel density in 68 cancer tissue, peritumoral tissue samples and 20 normal controls were studied by PCR-SSCP and immunohistochemical methods. RESULTS: The positive rate of p53 and K-ras gene mutation and the expression of VEGF in cancer tissue (47.1%, 32/68; 44.1%, 30/68; 55.9%, 38/68) were significantly higher than in peritumoral tissue (13.2%, 9/68; 7.4%, 5/68; 11.8%, 8/68). p53, K-ras gene mutation and the expression of VEGF were not detected in 20 normal tissue. The expression of VEGF was closely related with the angiogenesis and metastasis of colorectal carcinoma (r = 0.820, P < 0.01). VEGF expression correlated with both p53 and K-ras gene mutation (P < 0.01). CONCLUSIONS: p53 and K-ras gene upregulated the expression of VEGF. p53 and K-ras gene might play an important role in modulating tumor angiogenesis. PMID- 12783700 TI - [A study of diagnosis of primary biliary reflux gastritis]. AB - OBJECTIVE: To study the diadynamic method of primary bile reflux gastritis. METHODS: Endoscopy, histologic examination, symptom analysis and 24-hour gastritic bilirubin monitoring with Bilitec 2000 were performed in 20 healthy subjects (HS) and 42 patients with symptoms of abdominal pain, abdominal distention, nausea and bile vomiting. RESULTS: The total fraction time of bile reflux was higher in 42 patents than in 20 HS [(17.68 +/- 17.89)% vs. (2.92 +/- 2.39)%, P < 0.01]. 23 (55%) of 42 patients showed pathologic duodenogastric reflux (DGR). The incidence of pathologic DGR detected by Bilitec2000 in bile dyed mucous lake, antral mucosal erosion, bile dyed mucous lake accompanied with erosion and bile dyed mucous lake accompanied with middle-hyperemia of antral mucosae found in endoscopy were 86%, 88%, 8/8 and 85% respectively. The diagnosis of bile reflux gastritis (BRG) could be established if the patients with bile dyed mucous lake accompanied with erosion and/or middle-hyperemia of antral mucosae were found to be pathologic DGR. With this criterion, 11 patients were BRG in 42 patients in this study. The active inflammation of antral mucosae in the patients with BRG that without Helicobacter pylori (Hp) infection was more severe than that in the patients with physiologic DGR that without Hp infection significantly (P < 0.05). The symptoms of abdominal distention, nausea and bile vomiting in the patients with BRG were more severe significantly than those in the patients with physiologic DGR respectively (P < 0.05). CONCLUSIONS: Final diagnosis of BRG can be made for the integrity stomach if the patients with bile dyed mucous lake accompanied with erosion and/or middle-hyperemia of antral mucosae were found to be pathologic DGR by Bilitec 2000. PMID- 12783701 TI - [Experimental study of apoptosis and proliferation of gastric mucosal cells in chronic atrophic gastritis]. AB - OBJECTIVE: To study the relationship between long-term high-salt hot diet and chronic atrophic gastritis (CAG). The expression of apoptosis cells, proliferation cells, bcl-2 and Fas proteins were detected in the gastric mucosa of CAG rates induced by high-salt hot water. METHODS: TUNEL technique and immunohistochemical method were used in the experiment. Flow cytometry method was used to detect the DNA content of apoptosis cells. RESULTS: (1) The detection of apoptosis with TUNEL showed that the apoptosis cells in CAG rates induced by high salt hot water were high. These cells could be seen in all layers of gastric mucosa, apoptosis index of model rates was higher than that of normal rates (P < 0.05). (2) The result of detecting apoptosis gene and proliferating cell nuclear antigen (PCNA) by immunohistochemical method showed that the expression of bcl-2, Fas and PCNA in the CAG rates induced by high-salt hot water was much higher than that of normal gastric mucosa (P < 0.05). The expression rates of bcl-2, Fas proteins were higher with time going on. After being given high-salt hot water intragastrically for 4, 8, 12 weeks, the bcl-2 protein expression rates were 12.5%, 16.7%, 76.5%, the Fas protein expression rates were 18.7%, 22.2% and 64.7%. (3) Flow cytometry technique showed that there was a second G(0)/G(1) peek, which was apoptosis cells peak, but was not seen in the normal gastric mucosa. CONCLUSIONS: Abnormality of proliferation and apoptosis in gastric mucosa could be induced by high-salt hot water. bcl-2, Fas gene played an important role in this course. PMID- 12783702 TI - [A randomized, double-blind, placebo-controlled, multicenter study to assess the efficacy and safety of tegaserod in Chinese patients with constipation predominant irritable bowel syndrome]. AB - OBJECTIVE: To determine the efficacy and safety of tegaserod 6 mg b.i.d. in the treatment of constipation-predominant irritable bowel syndrome (C-IBS). METHODS: An 8-week, double-blind, randomized, parallel group, placebo-controlled, multicenter study in 510 Chinese patients who met the Rome II criteria for C-IBS. The study consisted of a 2-week baseline period, a 4-week randomized, double blind treatment period with either tegaserod 6 mg b.i.d. or placebo (tegaserod:placebo = 1:1), followed by a 2-week withdraw period. Efficacy was assessed by patient's perception of overall symptoms of IBS during the previous week and severity of the patient's constipation during the last week and patient's assessment of their individual IBS symptoms. Safety was assessed by adverse events, laboratory evaluations, blood pressure and heart rates, Physical examination and ECG evaluations. RESULTS: The weekly severity of patients' perception of overall IBS symptoms was significantly lower in the tegaserod group from week 1 onwards and throughout the treatment period. The effects of tegaserod on secondary IBS efficacy parameters were consistently better in the tegaserod group starting in week 1 and lasting throughout the treatment period and withdraw period. Regarding adverse events during the treatment period, about 10% of the patients in the tegaserod group experienced an adverse event compared to 6% in the placebo group. Diarrhea, abdominal pain and dizziness were more frequent in the tegaserod group but had a low frequency. No serious adverse was observed due to tegaserod. CONCLUSIONS: Tegaserod 6 mg b.i.d. was effective in relieving overall IBS symptoms, constipation, abdominal discomfort and pain, bloating, etc with significant effects starting in week 1 and continuing throughout the treatment period. Tegaserod was generally well-tolerated and has no clinically relevant safety findings. PMID- 12783703 TI - [The safety and efficacy of pre-treatment with enoxaparin (Clexane) in acute coronary syndrome patients undergoing percutaneous coronary intervention]. AB - OBJECTIVE: The purpose of the study is to prospectively investigate the safety and efficacy of anticoagulation with enoxaparin (Clexane) in UA/NQMI patients undergoing PCI procedures. METHODS: UA/NQMI patients received Clexane 1 mg/kg, subcutaneously, q12h for at least 48 hours, the coronary angiography was immediately followed by PCI when indicated and is performed within 8 h after the morning injection. No additional UFH or LMWH was used during or after PCI. Blood samples were taken for further measurement of the anti-Xa activity in 176 patients. RESULTS: 507 UA/NQMI patients were included in the study. 176 patients (93.2%) of the average anti-Xa activity value was > 0.5 IU/ml. During follow-up within 30 days, 3.2% of the patients experienced AMI and 6.7% of the patients recurrent UAP. One patient (0.2%) received revascularization and another died of duodenum perforation. The rate of minor hemorrhage was 4.7% (24 patients). In 30 days follow up, one experienced NQMI and 1 recurrent UA among angiography patients. CONCLUSION: Subcutaneous Enoxaparin given at least for 48 hours before PCI with out additional UFH or LMWH during or after PCI was both safe and effective in high risk UA/NQMI patients. PMID- 12783704 TI - [The incidence of ischemic and hemorrhagic stroke in Chinese populations]. AB - OBJECTIVE: To study the ratio of ischemic to hemorrhagic stroke incidence in Chinese populations. METHODS: Fifteen populations in China, each including about 100,000 residents, were monitored from 1991 to 2000 for the occurrence of stroke. Stroke was classified as hemorrhagic, ischemic and unclassified based on the clinical data and CT scan results respectively. RESULTS: The average CT scan rate in stroke patients from 1996 to 2000 varied from 14.8% to 97.5% among the 15 population and the average ratio of ischemic to hemorrhagic stroke ranged from 0.23:1 to 4.38:1; the correlation coefficient between the ratio and CT scan rate is 0.7 (P = 0.003). In all the populations with CT scan rate greater than 80%, the ischemic to haemorrhagic ratios were all greater than 1.0. From 1991 to 2000 in 12 populations with complete data, the CT scan rate in stroke patients rose from 41.1% to 88.3% and the ischemic to hemorrhagic ratio rose from 1.25:1 to 1.85:1. The correlation coefficient between the ratio and the CT scan rate was 0.9 (P < 0.001). In patients having CT scan, the ischemic to hemorrhagic ratio was greater than 1.0 in all the years. CONCLUSION: The predominant type of stroke in Chinese population is ischemic. The low CT scan rate in some populations is the main reason for the false impression predominantly hemorrhagic stroke. PMID- 12783705 TI - [The effects of weight reduction in reversing fatty liver changes in overweight and obese patients]. AB - OBJECTIVE: To study the effects of weight loss on nonalcoholic steatohepatitis (NASH) in obese patients. METHODS: It is a prospective study. Blood biochemical parameters were examined in 220 overweight or obese patients without alcoholic consumption and without detectable HBsAg. Ultrasonographic examination was performed by a fixed doctor. 45 obese patients followed a program of weight loss, including diet restriction, exercise or drug for a trial period of one year. Weight, blood biochemical tests and ultrasonic examination of liver were compared before and after the trial in all the patients. RESULTS: 75.9% of the patients were diagnosed as NASH. Logistic regression analysis revealed that the risk factors of NASH were body mass index (BMI, beta = 0.926, P = 0.021), age (beta = 0.973, P = 0.021), 2-hour postprandial glucose (beta = 0.987, P = 0.012), 1-hour postprandial net insulin in serum (beta = 1.027, P = 0.032), waist circumference (beta = 1.091, P = 0.038)and low density lipoprotein cholesterol (beta = 0.980, P = 0.041). After the treatment, 65.7% of the patients with NASH improved. The level of ALT in these patients decreased significantly (P = 0.040). The improvement of NASH correlated positively with the reduction of BMI (beta = 3.032, P = 0.010), triglycerides (TG, beta = 1.041, P = 0.025) and waist circumference (beta = 1.115, P = 0.029). The decrease of ALT level correlated with the decrease of BMI (beta = 1.165, P = 0.002), TG (beta = 0.986, P = 0.005), waist circumference (beta = 0.736, P = 0.041), and the increase of high density lipoprotein cholesterol (beta = -0.772, P = 0.016). CONCLUSION: NASH is one of the most important complications of obesity. Weight loss is the mainstay of treatment for obese patients. PMID- 12783706 TI - [The expression of G proteins alpha-subunit mRNA in different thyroid diseases]. AB - OBJECTIVE: To investigate the expression of G proteins alpha subunit mRNA in different thyroid diseases. METHODS: Twenty-three thyroid specimens were obtained during surgery, 5 from patients with Graves' disease (GD), 7 from patients with multinodular goiter (MNG), 6 from patients with eufunctioning thyroid adenoma (EFTA) and 5 from patients with thyroid papillary cancer (TPC). The expression of stimulating and inhibiting G proteins alpha subunit mRNA were determined by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: (1) The expression of G(s)alpha mRNA in TPCs (1.67 +/- 0.25) was significantly higher than that in normal thyroids (1.10 +/- 0.14) and MNGs (0.96 +/- 0.31), P < 0.05 and P < 0.01. The expression in GDs (1.47 +/- 0.11) and EFTAs (1.36 +/- 0.28) was significantly higher than that in MNGs (P < 0.05), but comparable to that in normal thyroids. (2) The expression of G(i)alpha-2 mRNA in GDs (0.68 +/- 0.26) was lower than that in MNGs (1.15 +/- 0.35), P < 0.05, but there was no difference in the expression of G(i)alpha-1 and G(i)alpha-3 mRNA among different thyroid diseases. CONCLUSION: The results indicated that G(s)alpha could play an important role in the pathogenesis of thyroid papillary cancer and G proteins had different expression in different thyroid diseases. PMID- 12783707 TI - [Clinical study on long-term treatment of primary pulmonary hypertension with continuous intravenous epoprostenol infusion]. AB - OBJECTIVE: To observe the effects of treatment with long-term continuous epoprostenol (Epo) infusion on the survival and pulmonary artery pressure in patients with primary pulmonary hypertension (PPH). METHODS: Sixty nine patients with PPH and congestive heart failure of stage III or IV (classification by New York Heart Association) treated with Epo of initial dose of 2 - 62 ng x kg(-1) x min(-1) (the dosage was enhanced by 1 - 2 ng x kg(-1) x min(-1) every 2 months according to patient's response). All patients underwent right heart catheterization and Doppler echocardiography to measure the systolic pressure gradient between the right ventricle and right atrium (DeltaP) and the cardiac output (CO). Doppler echocardiography and catheterization data were compared. Patients were followed up with echocardiography once every 4 months. RESULTS: Of 69 patients treated with Epo 17 were followed up for > 330 days. During this period, DeltaP decreased from (84.1 +/- 24.1) mm Hg to (62.7 +/- 18.2) mm Hg (P < 0.01), CO increased from (4.00 +/- 1.22) L/min to (4.70 +/- 1.27) L/min (P < 0.02), DeltaP/CO decreased from 22.8 +/- 9.4 to 14.9 +/- 6.5 (P < 0.01). The patient survival was improved. CONCLUSIONS: Patients with PPH treated with continuous infusion of Epo experienced a decrease in pulmonary artery pressure and an increase in cardiac output. Long-term follow-up of some patients of the group demonstrated improved survival during the period of Epo therapy. PMID- 12783708 TI - [Chinese herbs-induced renal failure with Fanconi syndrome: a report of 6 cases]. AB - OBJECTIVE: To understand the clinical and pathological features of patients with Fanconi syndrome associated with renal function damage induced by Chinese herbs. METHODS: Six cases with herb-induced renal failure associated with Fanconi syndrome were clinicopathologically analyzed. RESULTS: All six patients had kidney insufficiency after ingestion of Chinese herbs. All of them took the herbs containing aristolochia manshuriensis definitely. Four of the six presented with rapidly progressive acute renal failure and one with acute renal failure. All of them had similar clinical features, such as polydipsia, polyuria, anemia, glycosuria, aminoaciduria, increased urine beta(2) microglobin (beta(2)m) excretion and proximal tubular acidosis. Renal biopsies performed in 3 cases showed extensive hypocellular interstitial fibrosis, tubular atrophy and loss of tubules. Glomeruli were apparently intact. CONCLUSIONS: Nephropathy caused by Chinese herbs may be associated with Fanconi syndrome and renal failure. PMID- 12783709 TI - [Effect of arsenic trioxide on the expression of cyclins gene in HL60 cells]. AB - OBJECTIVE: To investigate the expression of cyclins related gene in HL60 cells before and after arsenic trioxide treatment by gene chip. METHODS: Total mRNAs were extracted from untreated or arsenic trioxide treated HL60 cells. Then two cDNA probes were made from these two mRNAs which were labeled by fluoro link Cy3 ducpp deoxyuridine triphosphate (Cy3-dUTP) or Cy5-dUTP fluorescence dyes respectively, hybridized with gene chip and scanned for fluorescent intensity. Different expression genes were then screened out. Cell apoptosis was detected by electron microscopy, in site cell apoptosis detection kit, DNA agarose gel electrophoresis and flow cytometry (FCM). RESULTS: Among the 82 genes which expressed differently after treatment with arsenic trioxide, 34 genes were up regulated, while 48 genes down-regulated. It was detected that 15 micro mol/L As(2)O(3) can definitively induce HL60 cells to go apoptosis by FCM. Rate of apoptotic HL60 cells in control group is 1.7%, and As(2)O(3) group is 26.1%. CONCLUSION: Cyclin B1, proliferating cell nuclear antigen (PCNA), insulin like growth factor binding protein (IGFBP) et al may play an important role in HL60 cell apoptosis induced by arsenic trioxide. PMID- 12783710 TI - [Diabetic retinopathy and its risk factors]. PMID- 12783711 TI - [Study of the relationship between retinal and renal microangiopathy in patients with diabetes mellitus type 1]. AB - The similarities in the vascularization of the retina and the kidney give rise to the complications of diabetes mellitus in the small vessels (microangiopathy) that appear in both organs. The present study presents the prospectively obtained results along 10 years in 104 diabetic type 1 patients, with the characteristics not to present microalbuminuria nor diabetic retinopathy at the beginning of the study; the results bear as objective the establishment of the relation between the presence of proliferative diabetic retinopathy and microalbuminuria, taking into account different factors of epidemiological risk: gender, duration of diabetes, presence of diastolic hypertension, levels of HbAlc, and blood lipids. The results obtained indicate a 10 years of follow-up incidence of diabetic retinopathy of 37.50% (being the proliferative form 13.46%) and the incidence of microalbuminuria is of 20.19%. The appearance of proliferative diabetic retinopathy is associated to the presence of diastolic hypertension, while the appearance of microalbuminuria is related to the elevation of levels of HbAlc. The results obtained in the proliferative diabetic retinopathy and microalbuminuria incidence are similar to those of other authors. At the end of the study four groups of patients are configured depending on whether they present proliferative diabetic retinopathy and/or microalbuminuria. The results obtained in the present study suggest the possible existence of two populations of diabetic patients, one that would develop proliferative diabetic retinopathy and renal injury, and a second population in which only would appear proliferative diabetic retinopathy. PMID- 12783712 TI - [Clinical manifestations and level of blood pressure in patients evaluated for the first time in a Hypertension Clinic]. AB - OBJECTIVE: To analyze the clinical manifestations, blood pressure level, and determinants of the degree of hypertension (HT) control in the patients referred by the Primary Care physicians and evaluated for the first time in a Hypertension Clinic. METHOD: The participants were 1,317 consecutively referred patients for the first time to the Hypertension Clinic by their Primary Care physician, between January 1997 and November 2000. The following initial study protocol was applied: case history and physical exploration, blood and urine biochemistry, chest and abdomen x-rays, fundus oculi and electrocardiogram. According to the initial clinical orientation and the results of the complementary tests described, the study was supplemented or not with other complementary studies. Results were registered in a computer dabatase and subsequently were analyzed with the statistical program SPSS 9.0. RESULTS. 741 (56.3%) of the 1,317 patients were women. The average age was 54.58 14.4 years (range: 14-91 years). Average systolic blood pressure (SBP) and diastolic blood pressure (DBP) were 152.6 22.7 mmHg and 94.2 11.6 mmHg, respectively. 71.6% (943 patients) presented at least two cardiovascular risk factors (CVRF), 50.5% (665 patients) three CVRF and 10.4% (137 patients) four CVRF. 43.6% (575 patients) presented a degree I, 33.2% (438 patients) a degree II and 23.1% (304 patients) a degree III HT. The degree of HT was associated with the age, the time elapsed from the beginning of the HT, the body mass index (BMI) and the levels of blood glucose, total cholesterol and LDL cholesterol. A model of logistic regression to classify the patients as HT degrees I or II-III identified as independent variables the age, the time elapsed from the beginning of HTA, the BMI, the presence of diabetes mellitus (DM) and the plasma LDL cholesterol level greater of 160 mg/dl. CONCLUSIONS: Half of the patients sent for the first time by its Primary Care physician to our Hypertension Clinic presented at least three CVRF. Furthermore, 56.4% presented a moderate-severe HTA (degrees II-III). Finally, the degree of HTA of the patients was related to the age, the time elapsed from the beginning of the HTA, the Quetelet's index, and the existence of DM and hypercholesterolemia (HC). PMID- 12783713 TI - [Recurrent tuberculosis in patients with coinfection by HIV]. AB - OBJECTIVES: Analysis and identification of factors associated with the tuberculosis (TB) recurrence in HIV-infected patients. PATIENTS AND METHODS: The HIV-infected patients diagnosed with TB between 1995-2000 and with correct completion of the treatment were selected. There were compared those patients who presented a tuberculous disease recurrence with those which did not presented it. Multivariate analysis was carried out by logistic regression in order to identify factors associated with a new episode of TB. RESULTS: In the total sample of 223 diagnosed HIV-infected patients with TB, 159 (71.3%) patients completed adequately the treatment and were considered cured. The rest were excluded because of: death before completing the treatment (5.8%), lost in the follow-up (11.7%), therapeutic abandonment (9.4%), and therapeutic failure (1.8%). In the 159 patient final sample, 14 (8.8%) patients presented later recurrence. The patients with recurrent TB presented a greater degree of immunosuppression, more previous complications indicative of AIDS, and greater frequency of extrapulmonary TB in the initial episode. After the multivariate analysis, a count of lymphocytes CD4 lower than 100/mm3 in the first episode of TB was associated with recurrent tuberculous disease (odds ratio [OR]: 4,6; 95% confidence index [CI]: 1,3-18,2). Mortality was high (35.7%) in the patients with recurrent episodes. CONCLUSIONS: The recurrence of TB in patients coinfected by HIV occurs in individuals with profound immunossuppression and is associated to high mortality. PMID- 12783714 TI - [Community-acquired Acinetobacter baumannii pneumonia]. AB - OBJECTIVES: Description of two cases of community-acquired Acinetobacter baumannii pneumonia. PATIENTS AND METHODS: Two patients, one with a history of diabetes mellitus type 2 and pulmonary thromboembolism 2 years earlier and the second without known risk factors except for the age, were admitted to hospital because of community-acquired A. baumannii pneumonia with cavitation and with favorable final evolution. CONCLUSION: A. baumannii is an infrequent cause of community-acquired pneumonia, although it should be suspected in patients with debilitating illness and in patients who do not evolve favorably on the first days of the admission with conventional treatment. PMID- 12783715 TI - [Idiopathic hypertrophic chronic pachymeningitis. Contribution of two new cases and literature review]. AB - The idiopathic hypertrophic chronic pachymeningitis (IHCP) is a rare disorder characterized by chronic inflammation of the dura mater that there can be associated with resulting neurological deficits from the compression of underlying structures. The etiology of the pathologic process is uncertain and for its diagnosis it is necessary to rule out infectious pathology, diseases of the immune system and other processes, together with the pathological confirmation of the existence of a steril and chronic inflammatory infiltrate of the dura mater, and without evidence of vasculitic sings or malignancy. The IHCP is an infrequent entity. Through Medline last 35 years review we have found 58 cases of IHCP, of which in 43 the affectation was located at intracranial level, being this the most frequent localization, followed by 11 cases where the affectation was located at spinal level and 4 at craniospinal level. We present two clinical cases of IHCP of spinal localization, one at dorsal level and another one at the lumbar level seen in our institution, and we review the clinical manifestations, diagnostic methods, and therapeutic attitude used in the cases found in the literature. PMID- 12783716 TI - [Fabrys disease: a multidisciplinary entity with new therapeutic insights]. PMID- 12783717 TI - [National Health System: equity versus efficiency]. PMID- 12783718 TI - [Adherence to the antiretroviral treatment]. PMID- 12783719 TI - [Polyarthritis and primary biliary cirrhosis]. PMID- 12783720 TI - [25 year-old male with a worm in the vomit and facial swelling]. PMID- 12783721 TI - [Fever and cutaneous lesions in leg in a 20 month-old girl]. PMID- 12783722 TI - [Microscopic polyangiitis: contribution of 5 cases]. PMID- 12783723 TI - [Amyotrophic neuralgia (Parsonage-Turner syndrome) with bilateral presentation in a patient operated on account of surgical abdomen]. PMID- 12783724 TI - [Relapsing liver abscesses by Klebsiella pneumoniae]. PMID- 12783725 TI - [Mammary, cutaneous and nodal tuberculosis without pulmonary affectation in patient without apparent immunosuppression]. PMID- 12783726 TI - [Role of echocardiography in the detection and prognosis of ischemic mitral regurgitation]. AB - Ischemic mitral regurgitation (IMR) is mitral regurgitation (MR) due to complications of coronary artery disease and not fortuitously associated with it. Acute MR secondary to ruptured papillary muscle after myocardial infarction is rare and often fatal. We focus on functional MR, much more common, which occurs without any intrinsic valve disease. It was often underrated because of low murmur intensity but with the use of echocardiography this complication is observed between 15%-20% after a myocardial infarction. Recent advances in noninvasive Doppler echocardiography allow accurate assessment of regurgitant volume and effective regurgitant orifice and thus provide the tools to reliably evaluate the prognosis and mechanisms of IMR. IMR presence is associated with excess mortality. The mortality risk is directly related to the degree of regurgitation and a regurgitant volume > or = 30 ml or an effective regurgitant orifice > or = 20 mm2; define a high-risk group. Presence and degree of the regurgitation are related to local left ventricular remodeling. The apical and posterior displacement of papillary muscles leads to excess valvular tenting which in turn, in association with loss of systolic annular contraction, determines the severity of the regurgitation. In current clinical practice, IMR is mainly corrected by ring annuloplasty. However, this technique does not correct local alterations of left ventricular remodeling and its benefits on long term outcome remains to be demonstrated. PMID- 12783727 TI - [The changing prognosis of myocardial infarction in the reperfusion era: implications for evaluation and management of ventricular arrhythmias]. AB - The prognosis of patients with ischemic heart disease has improved markedly with the introduction of reperfusion therapy and with aggressive efforts to modify risk factors. Consequently, the rate of cardiovascular events after myocardial infarction has decreased to approximately 5% over a period of 2 years as compared with the 20% to 30% reported in the prethrombolytic era. In this context, it is unlikely that the results of previous studies in the prereperfusion era can be applied to this group of patients. Others have demonstrated that the identification of subgroups of patients at greater risk and the search for new risk markers can significantly improve survival of patients who are at high risk despite reperfusion therapy. For example, it was found in the GISSI-2 study that unsuitability for exercise stress testing was associated with a mortality of 7% at 6 months. Other factors that determine poor prognosis after myocardial infarction are transitory heart failure, left ventricular dysfunction, and advanced age. The active search for new risk markers has identified other factors such as nonresolution of ST-segment changes, impaired ventricular filling, anomalous baroreflex sensitivity, or T-wave alternans that may be of benefit in assessing risk. Also, the timing of risk stratification can be critical. Often, risk factors have been analyzed weeks or even months after infarction instead of before hospital discharge. Approximately 30% of patients have deterioration of left ventricular function in the next 2 or 3 months, whereas others have improvement, highlighting the difficulties in attempting to risk stratify at one point in time. Although nobody doubts the effect that coronary revascularization has had on the prognosis of ischemic heart disease or the effectiveness of aspirin, beta -blockers, lipid-lowering drugs, and angiotensin-converting enzyme inhibitors, the search for cardiac or noncardiac risk markers can contribute notably to increasing the survival of patients who have had myocardial infarction. PMID- 12783728 TI - [Current challenges in infective endocarditis]. PMID- 12783729 TI - [Endothelial dysfunction and intima-media thickness in relation to cardiovascular risk factors in patients without clinical manifestations of atherosclerosis]. AB - INTRODUCTION: Endothelial dysfunction and increased intima-media thickness are early findings in the development of atherosclerosis that can be assessed non invasively by echography. The aim of this study was to investigate endothelial function and intima-media thickness, and the relation between these processes and cardiovascular risk factors in patients without clinical atherosclerosis. PATIENTS AND METHOD: Fifty-two subjects were studied, 39 with one or more cardiovascular risk factors and 13 with none. Vascular echography was performed to analyze endothelium-dependent vascular dilatation in the brachial artery and intima-media thickness in the common carotid artery. RESULTS: Compared to patients without risk factors, patients with cardiovascular risk factors more frequently had impaired vascular dilatation after ischemia, 11.98 4.61% vs 2.77 2.57%, (P<.0.001; mean difference = 9.21%, 95% CI of the difference 6.33-12.07%) and a greater intima-media thickness, 0.085 0.024% vs 0.057 0.014 cm (P < 0.0001; mean difference = 0.028 cm, 95% CI of the difference, 0.017-0.04 cm). There was a significant negative correlation between intimal-media thickness and endothelial dysfunction (r = -0.357; P<0.01). Linear regression analysis showed that intima media thickness was independently related to age and the presence of hypertension, while endothelial function was related only with the presence of hypertension, smoking, and hyperlipoproteinemia. CONCLUSIONS: In patients without clinical atherosclerotic disease, cardiovascular risk factors were associated with impaired endothelial function and increased intima-media thickness. There was a negative correlation between endothelial-dependent vascular dilatation and intima-media thickness. PMID- 12783730 TI - [Circadian variations in proinflammatory cytokine concentrations in acute myocardial infarction]. AB - INTRODUCTION AND OBJECTIVES: The concentration of certain proinflammatory cytokines has been found to be elevated in patients with acute coronary syndrome. Many studies have shown that coronary ischemic accidents do not show a uniform distribution throughout the day, but instead exhibit rhythmic variations. The objective of this study is to determine whether there is a circadian pattern of variation in the concentrations of proinflammatory cytokines in patients with acute myocardial infarction. PATIENTS AND METHOD: The sample included 40 patients with acute myocardial infarction and 40 controls. Levels of interleukin 6 and 1 beta were determined in the first 24 hours after the acute coronary ischemic episode. Blood samples were extracted at 3:00 a.m. (period of darkness) and at 10:00 a.m. (period of daylight). RESULTS: Both groups were similar in age, sex distribution, and coronary risk factors. Interleukin 6 levels showed a significant variation between daylight and nighttime concentrations in patients with acute myocardial infarction and controls (41.93 5.90/100.39 13.60 vs 25.76 4.45/52.67 7.73 pg/ml). However, interleukin 6 concentrations were higher in the acute myocardial infarction group than in the control group. Interleukin 1 beta concentrations did not vary between daylight and darkness. CONCLUSIONS: In both the control group and acute myocardial infarction group, interleukin 6 concentrations varied between daylight and darkness. Patients with acute myocardial infarction shown a higher concentration of interleukin 6 secondary to the physiological response to tissue damage. Circadian variations can affect the measurements obtained for different physiological and biochemical parameters. PMID- 12783731 TI - [Detection of angiographic lesions in the left anterior descending coronary artery by transthoracic Doppler echocardiography: usefulness of non-invasive assessment of coronary flow reserve]. AB - INTRODUCTION: We evaluated the feasibility of detecting blood flow in the left anterior descending coronary artery and the usefulness of measuring coronary flow reserve to diagnose significant coronary artery disease, both by means of transthoracic Doppler echocardiography using a high-frequency transducer and echo contrast agent. PATIENTS AND METHOD: We studied 107 patients who were scheduled for coronary arteriography for known or suspected ischemic heart disease. A Doppler signal was recorded by a pulsed wave in the distal left anterior descending artery at baseline and after dipyridamole infusion. An echo-contrast agent was administered to all patients. A coronary flow reserve equal to or higher than 1.7 was considered normal. RESULTS: We recorded Doppler signals in the left anterior descending coronary artery of 83 patients (78%). Significant stenosis of the left anterior descending coronary artery was observed in 24 out of 83 patients (29%). The prevalence of significant stenosis was higher (62 vs 29%; p = 0.006) in patients in which no Doppler signal was detected. The sensitivity, specificity, and accuracy of abnormal coronary flow reserve in detecting significant stenosis of the left anterior descending coronary artery were 87, 74 and 78%, respectively. CONCLUSIONS: The measurement of coronary flow reserve by transthoracic Doppler echocardiography using a high-frequency transducer and echo-contrast agent is a feasible, widely available, and accurate method for detecting significant stenosis of the left anterior descending coronary artery. PMID- 12783732 TI - [Complications with femoral access in cardiac cathetization. Impact of previous systematic femoral angiography and hemostasis with VasoSeal-ES collagen plug]. AB - INTRODUCTION AND OBJECTIVES: Most cardiac catheterizations are performed via femoral artery access, and hemostatic devices are commonly used. We evaluate the relationship between the strategy used for femoral arteriography and the use of VasoSeal-ES, and local vascular complications. PATIENTS AND METHOD: Prospective study of 540 consecutive catheterizations with systematic femoral artery and sheath angiography. VasoSeal-ES was used in 427 patients. Predictors of local vascular complications such as patient-related factors, anatomy and hemostasis were analyzed. Variables related to failure of the collagen plug were also studied. RESULTS: Punctures of the common femoral artery occurred in 35.9% of all patients (16% in the deep femoral artery and its ostium). Spasm was evident in 18% (ranging from 58.1% in the deep femoral artery to 5.2% in the common femoral artery). Puncture at the site of ramification was seen in 11.3%. Angiographically significant atheroma was seen in 17.8%. The femoral head was a valid landmark for the common femoral artery in only 63.9% of the pateints. Risk factors for local vascular complications were punctures of the common femoral artery, female sex and failure of VasoSeal-ES to achieve hemostasis (15.8% in the first two months of use, 5.2% in the last months of the study). Complications involving superficial and deep femoral arteries occurred in 6.7% and 1.2% of the patients, respectively, in contrast to 0.6% involving the common femoral artery. Variables related to collagen plug failure were patient-related factors, weight less than 55 kg, operator-related factors and the learning curve. CONCLUSIONS: Systematic femoral angiography provides data that aids the choice of the best hemostasis procedure to reduce local vascular complications. Punctures of the common femoral artery were more frequent than expected, and were associated with a higher complication rate. VasoSeal-ES is a safe and useful method of hemostasis, and its infrequent failures were associated with high complication rates that were substantially reduced with experience. PMID- 12783733 TI - [Predictors of hospital mortality in 186 cases of active infective endocarditis treated in a tertiary medical center (1992-2001)]. AB - INTRODUCTION AND OBJECTIVES: The aim of this study was to describe the predictors of hospital mortality found in patients admitted for infective endocarditis (IE) to a cardiovascular surgery ward. Patients and method. Prospective study of 186 patients with IE treated in our hospital between 1992 and 2001. RESULTS: One hundred fourteen patients (61.3%) had native valve endocarditis and 72 (38.7%) had prosthetic valve endocarditis (early in 28 patients [up to 12 months after surgery] and late in 44 [later than 12 months]). Blood cultures were positive in 82%. The predominant organism was Streptococcus viridans (36%) in native valve endocarditis and Staphylococcus aureus (33%) in prosthetic valve endocarditis. The hospital mortality was 22.6%. Severe sepsis (4.8%) produced a high mortality rate (88%) and was caused by Staphylococcus aureus in 60%. One hundred nineteen patients (64%) required surgery, 79 (66.4%) of them urgently. Negative blood cultures predicted need for surgery in native valve endocarditis (p < 0.05). The surgical mortality was 21.8% and was related to NYHA III-IV class (p = 0.014) and emergency surgery (p = 0.009) in patients with native valve endocarditis. This last factor also predicted higher surgical mortality in patients with early prosthetic valve endocarditis (p < 0.001). CONCLUSIONS: The hospital mortality of this group of patients with infective endocarditis treated in a tertiary medical center was high. The presence of severe sepsis, although infrequent, had a somber prognosis. Severe heart failure in native valve endocarditis and urgent surgery in native and prosthetic valve endocarditis increased surgical mortality. PMID- 12783734 TI - [Secondary prevention of coronary heart disease is less agressive in patients over 64 years]. AB - INTRODUCTION AND OBJECTIVES: Although elderly people has a higher incidence of coronary heart disease, this group is seldom included in clinical trials. Studies performed in Spain on elderly coronary heart disease patients have been conducted in hospital settings. The aim of our study was to analyse wether the management of coronary heart disease patients over 64 years of age cared by family physicians differed from that of the rest. PATIENTS AND METHOD: Cross-sectional multicentre study embedded in a clinical trial on 1,022 patients with stable coronary heart disease in which socio-demographic variables, comorbidity, treatment and cardiovascular risk- factor control were collected. RESULTS: Mean age was 64 10, 74.0% were men and 53.8% of subjects were over 64 years. Patients over 64 years had a greater cardiovascular comorbidity (87.7 vs 82.6%; p = 0.002) and received lower number of drugs than the rest in the prevention of recurrences (60.4 vs 70.9%; p < 0.001). Probability to receive less than two drugs on secondary prevention by subjects over 64 years was 0.45 (95% CI, 0.30-0.68) despite comorbidity, sex and cardiovascular risk profile.Conclusions. Coronary heart disease patients over 64 years receive less drugs for coronary event recurrence prevention than their younger counterparts despite their worse cardiovascular risk profile. PMID- 12783735 TI - [Cardiopulmonary function and exercise capacity in patients with morbid obesity]. AB - INTRODUCTION AND OBJECTIVES: The effect of obesity on cardiac function is still under discussion. The objective of this study was to assess cardiopulmonary capacity in morbidly obese patients. Patients and method. A symptom-limited cardiopulmonary exercise stress test was carried out in 31 morbidly obese patients (BMI 50 9 kg/m2) and 30 normal controls (BMI 24 2 kg/m2. Cardiovascular function was evaluated using the oxygen pulse (oxygen uptake/heart rate). RESULTS: There were no differences in age, sex and height between both groups. During the effort the obese subjects presented greater oxygen uptake, heart rate, systolic arterial pressure and minute ventilation and shorter test duration than control group (14 3 vs 27 4 min; p < 0.001). Oxygen pulse values were higher in obese patients. However, after oxygen uptake indexation by fat free mass, these differences disappeared, suggesting a similar cardiovascular function. At the end of the exercise, the control group reached 96% of their age-predicted maximal heart rate and their respiratory exchange ratio was 1 0.2. Obese patients only reached 86% and 0.87 0.2, respectively. CONCLUSIONS: Due to their need of more energy output to move total body mass morbidly obese patients have a reduced exercise capacity. They finish the test having done a submaximal exercise. However, during this effort they show a normal cardiopulmonar capacity. PMID- 12783736 TI - [Summary of the clinical studies reported in the 52nd annual Scientific Sessions of the American College of Cardiology (Chicago USA, March 30-April 2, 2003)]. PMID- 12783737 TI - [Evaluation of congenital heart disease in adults]. AB - Improvements in the diagnosis and surgical treatment of congenital heart disease during infancy and childhood have resulted in an outstanding increase in the prevalence of these entities during adulthood. Congenital heart disease in the adult represents a new diagnostic challenge to the consultant cardiologist, unfamiliar with the anatomical and functional complexities of cardiac malformations. Assessment of adult congenital heart disease with imaging techniques can be as accurate as in children. However, these techniques cannot substitute for a detailed clinical assessment. Physical examination, electrocardiography and chest x-rays remain the three main pillars of bedside diagnosis. Transthoracic echocardiography is undoubtedly the imaging technique which provides most information, and in many situations no additional studies are needed. Nevertheless, ultrasound imaging properties in adults are not as favorable as in children, and prior surgical procedures further impair image quality. Despite recent advances in ultrasound technologies such as harmonic or contrast imaging, other diagnostic procedures are sometimes required. Fortunately, transesophageal echocardiography and magnetic resonance imaging are easily performed in the adult, and do not require anaesthetic support, in contrast to pediatric patients. These techniques, together with nuclear cardiology and cardiac catheterization, complete the second tier of diagnostic techniques for congenital heart disease. To avoid unnecessary repetition of diagnostic procedures, the attending cardiologist should choose the sequence of diagnostic techniques carefully; although the information this yields is often redundant, it is also frequently complementary. This article aims to compare the diagnostic utility of different imaging techniques in adult patients with congenital heart disease, both with and without prior surgical repair. PMID- 12783738 TI - [Assessment of coronary by-pass grafts with multislice CT]. PMID- 12783739 TI - [Post-infarction ventricular septal defect treated during the acute phase by transcatheter closure with an Amplatzer septal occluder]. AB - A 75-year-old man had a large anterior myocardial infarction complicated by a ventricular septal defect, which was treated in the first 48 h by transcatheter closure using the Amplatzer septal occluder. Treatment was successful (with only mild residual post-procedure shunting) and coronary angioplasty with stent implantation at the point of occlusion of the middle left anterior descending artery was performed in a later intervention. However, the patient died 7 days after the procedure as a result of sepsis and ventricular failure. PMID- 12783740 TI - [Right atrial pheochromocytoma]. AB - Pheochromocytoma is a catecholamine-producing tumor and a rare cause of hypertension. Most cases are intra-adrenal and intrapericardial pheochromocytomas are extremely uncommon. We report the case of a 46-year-old woman with a 1-year history of hypertension, in which a right atrial pheochromocytoma was detected after a hypertensive crisis. 131I-metaiodobenzylguanidine scintigraphy and magnetic resonance imaging established the diagnosis. The tumor was successfully resected using cardiopulmonary bypass and the right atrium was reconstructed using bovine pericardium. PMID- 12783741 TI - [Acute myocardial dysfunction after nasal infiltration with cocaine]. AB - We report the case of a patient with severe myocardial dysfunction after nasal infiltration of cocaine during septoplasty. Complete recovery of myocardial function was observed in twelve days. Several reports have described chronic cardiomyopathy in long-term cocaine users, but only one case of acute cardiomyopathy. None of these cases were related to the medical use of cocaine. PMID- 12783742 TI - [Angiogenesis and statins, the highest the dose the best? ]. PMID- 12783743 TI - [Pharmacological interaction between flecainide and benziodarone]. PMID- 12783744 TI - [Management through integrated care processes: a necessary strategy]. PMID- 12783745 TI - [Prevalence of worker burnout and psychiatric illness in primary care physicians in a health care area in Madrid]. AB - AIM: To analyze worker burnout and the prevalence of psychiatric illness among primary care physicians, and to determine how burnout is related with sociodemographic and work-related factors. DESIGN: Cross-sectional, descriptive study. SETTING: Primary care centers in Area 8, Madrid (central Spain).Subjects. All 244 physicians (family medicine and pediatrics) who provided care at centers in Area 8 at the time of the study.Method. Anonymous, self-administered questionnaire that included the Maslach Burnout Inventory (worker burnout); GHQ 28 (possible mental illness); survey of sociodemographic and work-related factors, unhealthy behaviors, use of medications, and perceived quality of care and risk of medical errors in relation with work-related pressures; open question regarding causes of work-related stress. RESULTS: Response rate, 80.3%. Burnout was detected in 69.2% of the physicians (95% CI, 62.1%-75.4%) and was severe in 12.3%. The prevalence of possible psychiatric illness was 36.7%. Burnout was found to be related with possible psychiatric illness (P<.01); perception that work-related pressures frequently led to diminished quality of care and medical errors (P<.o1); permanent employment status (P<.05); more than 1700 patients on the patient list (P<.05); age between 37 and 46 years (P<.01) and patient load of 35 to 47 patients/day (P<.05). CONCLUSIONS: The prevalence of worker burnout and possible psychiatric illness was high, and the two variables were related. A large percentage of participants perceived that work-related pressures diminished the quality of care provided. PMID- 12783746 TI - [Commentary: chronic distress and worker burnout: hypotheses about causes and classification]. PMID- 12783747 TI - [Evaluation of Spanish websites useful for the primary care physicians]. AB - OBJECTIVE: To assess the usefulness of medical websites in spanish for the primary care physicians using quality criteria chosen by themselves. DESIGN: Observational, descriptive, cross-sectional study. SETTING: Internet. PARTICIPANTS: The search was performed in Google and Yahoo using as key words the name of Spanish National Health Service medical specialities, primary care and synonyms. The first one hundred links in spanish for every keyword were included in the study. EXCLUSION CRITERIA: paysites, commercial sites, non-clinical content, link sites, patient oriented sites and on-line editions of printed issues. METHODS: A survey was carried out asking the primary care physicians of area IV in Asturias which were the most important criteria that a medical website should fulfil. The results of this survey were used to make an assessment questionnaire. Every site was analysed randomizedly by two researchers and was classified as excelent, good, medium or bad. RESULTS: 2.095 medical websites in spanish were found, being repeated 1.218 (58.14%). From the 877 analysed websites, 597 (68.07%) were excluded. The Intraclass Correlation Coefficient (ICC) was 0.73 (95% CI, 0.66%-0.785%). 5 websites (1.8%) were rated as excelent and the 77.2% as medium or bad. The sites with better ratings were those oriented to the primary care professionals. CONCLUSIONS: The most websites were rated as medium or bad. The evaluation questionnaire is useful for futures studies. PMID- 12783748 TI - [Non-detected dementia and use of the health services: implications for primary care]. AB - OBJECTIVES: To calculate the proportion of cases of dementia detected in people over 70 living in their homes and to describe the use made by people with dementia of the health and social services. MATERIAL AND METHODS: Population survey of the survivors of the cohort "Growing old in Leganes", started in 1993. In the third monitoring (1999-2000), the clinical diagnosis of dementia on the basis of a neurological examination and an extensive neuro-psychological battery was included. Their use of health and social services and prior diagnoses were also asked. RESULTS: In the sample of survivors (n=527), there was 12.1% prevalence of dementia. Only 30% of the demented had previously been diagnosed by the health services. The proportion of undetected dementia was significantly associated with its seriousness (light 95%, moderate 69%, severe 36%). Compared with older persons who were not demented, the demented used more often hospital services, medical and nursing consultations at home and consultations through third parties; and less often, preventive and rehabilitation services. This trend was accentuated in patients with grave dementia. The use of community social services was very low (below 8% in the most serious cases). CONCLUSIONS: The detection of dementia in the elderly is very low and efforts to detect it in primary care need to be stepped up. Specific social-health resources for this population also need to be increased and the attendance guide-lines for primary care teams, and for health professionals in general, need to be changed. PMID- 12783749 TI - [Home care and family care in tackling the chronic illnesses of the elderly]. PMID- 12783750 TI - [Surveys as a research technique. Composition of questionnaires and statistical processing of data (II)]. PMID- 12783751 TI - [Immunization: leaps into the future. Combined vaccines (II)]. PMID- 12783752 TI - [Self-measurement of blood pressure in primary care (II)]. PMID- 12783753 TI - [Knowledge and degree of control of cardiovascular risk factors before and after a heart attack]. PMID- 12783754 TI - [Secondary prevention of ischaemic cardiopathy in a primary care center]. PMID- 12783755 TI - [Erythema dyschromicum perstans or Cinderella dermatitis. Concerning a case]. PMID- 12783756 TI - [Acute sarcoidosis: concerning a case]. PMID- 12783757 TI - [Co-infection of hepatitis C and HIV]. PMID- 12783758 TI - [Difficulties in the translation of pain adjectives: the case of breakthrough pain]. PMID- 12783759 TI - [Challenges we should meet in allergy and immunology]. PMID- 12783760 TI - [Hypersensitivity reactions to beta-lactam antibiotics in childhood]. PMID- 12783761 TI - [Special features of NSAID intolerance in children]. AB - Adverse reactions to acetylsalicylic acid (aspirin, ASA) and other non-steroidal anti-inflammatory drugs (NSAIDs) are the second most important cause of adverse drug reactions (ARDs) after beta-lactams. They produce various clinical manifestations and can affect different organs. Gastrointestinal reactions (pyrosis, vomiting, gastralgia), neurological reactions (tinnitus, deafness, vertigo), blood dyscrasias, and nephrotoxic and hepatotoxic reactions are well known.NSAIDs are the drugs of choice in the treatment of chronic arthropathies and other childhood connective-tissue diseases and are also commonly used in the treatment of febrile and acute inflammatory processes. Not all NAIDs are authorized for use in the pediatric population but their spectrum of use varies according to the entity for which they are indicated and the legislation of the country. Published studies on the prevalence of aspirin intolerance in patients with bronchial asthma show a fair amount of disagreement. This may be due to (i) the method of selecting asthmatic patients for the study, which differs according to whether all asthmatic patients are included or only those dependent on corticoids; (ii) the diagnostic method used, whether based on clinical criteria or oral provocation tests, which will affect the number of patients with a diagnosis of intolerance. In children aged less than 10 years, including children with asthma, the prevalence is low, while among children and young adults aged 10 20 years old, the prevalence is estimated at 10 %. Some hypotheses attempt to explain the mechanisms through which adverse reactions to NAIDs take place. One hypothesis attributes the reaction to a reaginic immunological mechanism but this hypothesis has only been confirmed in exceptional cases. The theory of the cyclooxygenase pathway, currently the most widely accepted, is based on the ability of NSAIDs to inhibit the cyclooxygenase pathway of arachidonic acid metabolism, leading to prostaglandin depletion and an increase in leukotrienes. The discovery of two isoforms of the cyclooxygenase enzymes, COX-1 and COX-2, has represented a great advance in our understanding of the mechanism of action of NSAIDs and has also elucidated the problem of cross-reactivities. According to the theory of viral infection, aspirin-induced asthma could be caused by chronic viral infection since, after initial exposure to the virus, cytotoxic lymphocytes are produced. Their activity is inhibited by prostaglandin E2 (PGE2); aspirin and other NSAIDs block PGE2 production and allow cytotoxic lymphocytes to attack and eliminate the respiratory tract cells infected by the virus. During this reaction lysosomal enzymes and mediators are released, which could precipitate an asthmatic crisis.Clinically, five types of reaction have been identified: 1. Respiratory illness with aspirin sensitivity. 2. Aspirin-induced urticarial disease. 3. Allergic reactions to NSAIDs and aspirin. 4 and 5. Aseptic meningitis and pneumonitis due to hypersensitivity. The latter are exceptional and are published as case reports. They have never been associated with aspirin or acetaminophen and usually occur in patients undergoing prolonged treatment. Diagnosis is based on a detailed history. Skin tests are not valid and in vitro tests are not widely used. Provocation tests with aspirin and NSAIDs definitively identify sensitized patients but their indications and limitations should be kept in mind. In children, certain features of adverse reactions to NSAIDs are observed in relation to their incidence and clinical manifestations. Acetaminophen is considered the drug of choice but further studies of other alternatives in children are required. PMID- 12783762 TI - [Adverse reactions to vaccines]. AB - Adverse reactions to vaccines are highly varied, ranging from mild local reactions to fatal outcomes. In the last few years many adverse reactions have been attributed to vaccines, often without justification. In agreement with the World Health Organization, these reactions can be classified as follows, depending on the cause: vaccination-induced reactions (due to an effect of the vaccine itself or to an idiosyncrasy); reactions due to errors in storage, manipulation and/or administration; and coincidental reactions (no causal relationship with the vaccine). Hypersensitivity reactions fall into six categories, depending on the causative agent: reactions due to some component of the infectious agent or one of its products; reactions due to adjuvants: aluminium hydroxide; reactions due to stabilizers: gelatin; reactions due to preservatives: thiomersal; reactions due to antibiotics: neomycin; and reactions due to a biological culture medium: chicken embryo cells. Allergic children should not be excluded from the normal vaccine calendar. Immunologically, allergic individuals are more susceptible to infection and to microbial and viral diseases, which often play an aggravating role. Rubella, whooping cough, and influenza usually exacerbate respiratory allergies. Non-vaccination carries a marked risk of contracting serious diseases such as poliomyelitis, tetanus, and diphtheria, etc. In a not too distant future, the techniques of genetic recombination and monoclonal antibody production will allow the creation of vaccines from organisms that cannot be cultivated in the laboratory or that produce small quantities of antigen. These techniques will also lead to identification of the antigens with the greatest immunogenic power and, consequently, to extremely pure vaccines. The adverse reactions to vaccines referred to our service account for between 0.59 % and 1.27 % of first visits in the last three years. We recorded a total of 48 adverse reactions to vaccines. Of these, 44 were attributed to the tetanus vaccine (92 %), 2 to the measles-mumps rubella vaccine (4 %) and 2 to the meningitis A and C vaccine (4 %). Clinical features consisted of urticaria (11 cases), urticaria with angioedema (7 cases), pseudo-shock (5 cases), fever and urticaria (4 cases), local reactions (4 cases), persistent crying with exanthema (3 cases), giant local reactions with angioedema of the limb (3 cases), anaphylaxis (3 cases), fever > 39.5 C (2 cases), bronchospasm (1 case), and severe atopic dermatitis (1 case).A regimen of hyposensitization to tetanus toxoid was required in 20 patients (45 %); in three, this could not be completed due to generalized urticaria but all the patients presented protective titers with diluted vaccine. PMID- 12783764 TI - [Cross-reactivity between fruit and vegetables]. AB - Vegetable foods are the most frequent cause of food allergy after the age of 5 years. The most commonly implicated foods are fruit and dried fruits, followed in Spain by legumes and fresh garden produce. In patients allergic to fruit and garden produce, multiple sensitizations to other vegetable products, whether from the same family or taxonomically unrelated, are frequent, although they do not always share the same clinical expression. Furthermore, more than 75 % of these patients are allergic to pollen, the type of pollen varying in relation to the aerobiology of the area. The basis of these associations among vegetable foods and with pollens lies in the existence of IgE antibodies against "panallergens", which determines cross-reactivity. Panallergens are proteins that are spread throughout the vegetable kingdom and are implicated in important biological functions (generally defense) and consequently their sequences and structures are highly conserved. The three best-known groups are allergens homologous to Bet v 1, profilins, and lipid transfer proteins (LTP). Allergens homologous to Bet v 1 (major birch pollen allergen) constitute a group of defense proteins (PR-10), with a molecular weight of 17 kDa, which behave as major allergens in patients from northern and central Europe with allergy to vegetables associated with birch pollen allergy. In these patients, the primary sensitization seems to be produced through the inhalation route on exposure to birch pollen. The symptomatology characteristically associated with sensitization to this family of allergens is oral allergy syndrome (OAS). Profilins are highly conserved proteins in all eukaryotic organisms and are present in pollen and a wide variety of vegetable foods. They have a molecular weight of 14 kDa and present a high degree of structural homology as well as marked cross-reactivity among one another. The presence of anti-profilin IgE broadens the spectrum of sensitizations to vegetable foods detected through skin tests and/or in vitro tests but whether it correlates with the clinical expression of food allergy is unclear.LTPs are the most commonly implicated allergens in allergy to Rosaceae fruits in patients from the Mediterranean area without birch pollen sensitization. LTPs are a family of 9kDA polypeptides, widely found in the vegetable kingdom and implicated in cuticle formation and defense against pathogens (PR-14). They are thermostable and resistant to pepsin digestion, which makes them potent food allergens and explains the frequent development of systemic symptoms (urticaria, anaphylaxis) in patients allergic to Rosaceae fruits in Spain. LTPs have also been identified in other vegetable foods and in pollens and a marked degree of cross-reactivity among them has been demonstrated, which may explain (together with profilin) the frequency of individuals sensitized to vegetable foods in the Mediterranean area. PMID- 12783765 TI - [Cross reactivity between fish and shellfish]. AB - In Spain, fish allergy represents 18 % of all cases of food allergy in children while reactions caused by crustacea and mollusks account for 3.8 % and 1.6 % respectively. Cross-reactivity is defined as the recognition of distinct antigens by the same IgE antibody, demonstrable by in vivo and in vitro tests, which clinically manifests as reactions caused by antigens homologous to different species. Subclinical sensitization can also occur, giving rise to patients sensitized to particular fish or shellfish but who do not present symptoms on consumption.Cod and shrimp have been the models used to study allergy to fish and crustacea respectively. The major allergens responsible for cross-reactivity among distinct species of fish and amphibians are proteins that control calcium flow in the muscular sarcoplasm of these animals, called parvalbumins, with a molecular weight of approximately 12 kD and an isoelectric point of 4.75, resistant to the action of heat and enzymatic digestion. Recently, recombinant carp parvalbumin has been reproduced, confirming that this allergen contains 70 % of the IgE epitopes present in natural extract of cod, tuna and salmon, which makes it a valid tool in the diagnosis of patients with fish allergy. Moreover, this recombinant allergen could constitute the basis for the development of immunotherapy against food allergy. In the case of shellfish, a non-taxonomic group that includes crustacea and mollusks, the major allergen is tropomyosin, an essential protein in muscle contraction both in invertebrates and vertebrates. In invertebrates, tropomyosins, which have a molecular weight of between 38 and 41 kD, show great homology in their amino acid sequence and are the panallergens responsible for cross-reactions between crustacea, insects, mites, nematodes, and different classes of mollusks. It is estimated that 50 % of individuals allergic to some type of fish are at risk for reacting to a second species, while those allergic to some type of crustacea present a risk of 75 % due to the greater similarity among tropomyosins than among parvalbumins. In addition, up to 40 % of patients sensitized to one or more fish do not present symptoms on consuming other species, the best tolerated of which belong to the Scombroidea family (which includes tuna). PMID- 12783766 TI - [Legume cross-reactivity]. AB - Legumes are dicotyledonous plants belonging to the Fabales order. The main distinctive characteristic of which is their fruit (legumen, seeds contained in pods). This botanical order is formed by three families: Mimosaceae, Caesalpiniaceae and Papilionaceae or Fabacea. The Papilionaceae family includes the most important allergenic species: Lens culinaris (lentil), Cicer arietinum (chick-pea), Pisum sativum (pea), Arachis hipogea (peanut), Phaseolus vulgaris (bean) y Glycine max (soy). Legumes are an important ingredient in the Mediterranean diet. Among Spanish children, sensitivity to legumes is the fifth most prevalent food allergy. Lentil and chick-pea are the most frequent cause of allergic reactions to legumes in Spanish children. Legumes could be involved in severe allergic symptoms. The different legumes have structurally homologous proteins, but they are not all equally allergenic, thus making it difficult to distinguish in vitro and in vivo cross-reactivity. We have demonstrated by skin tests and CAP that most of the patients are sensitised to more than one species. We have demonstrated a great degree of cross-reactivity among lentil, chick-pea, pea and peanut by ELISA inhibition (> 50 % max. inhibition). Unlike the Anglo Saxons population, this phenomenon implies clinical sensitisation for many Spanish children. The majority of our patients have had symptoms with more than one legume (median 3 legumes). Thirty-nine patients were challenged (open or simple blind) with two or more legumes and 32 (82 %) reacted to two or more legumes: 43,5 % to 3, 25,6 % to 2, 13 % to 4 legumes. Seventy three per cent of the patients challenged with lentil and pea had positive challenge to both, 69,4 % to lentil and chick-pea, 60 % to chick-pea and 64,3 % to lentil, chick-pea and pea simultaneously. Peanut allergy peanut can be associated to allergy to lentil, chick-pea and pea but less frequently. Contrarily, white bean and overall green bean and soy are well tolerated by children allergic to other legumes. In our study, 82 % of the children allergic to legumes had a sensitisation to pollen. Pea and bean are the legumes with more in vitro cross-reactivity with Lolium perenne, Olea europea and Betula alba. This cross-reactivity could be because of common antigenic determinants or due to the coexistence of pollen and legume allergy. Panallergens implication seems to be less probable. It is important to emphasize that in spite of an evident clinical and immunological cross reactivity, the diagnosis of legume allergy should not be based only on specific IgE tests. The decision to eliminate one legume from the diet should be based on a positive oral food challenge. PMID- 12783767 TI - [Bird-egg syndrome in children]. AB - Allergy to egg is highly frequent in childhood. In general, children allergic to egg react principally to the ingestion of egg white. Egg yolk contains various proteins but the major allergens are contained in egg white. The principal allergens are ovalbumin, ovomucoid, ovotransferrin, and lysozyme. These proteins have been sequenced. In some cases, a relationship between type I hypersensitivity with respiratory symptoms due to bird antigens and allergy to egg yolk has been described. This association is known as bird-egg syndrome, which is caused by sensitization to chicken serum albumin (alpha -livetin) and is characterized by the development of respiratory and gastrointestinal symptoms after egg intake or after contact with bird antigens. The initial symptoms are usually asthma with or without rhinoconjunctivitis due to contact with birds. Individuals first become sensitized to bird proteins (feathers, excrement, serum and meat) and subsequently develop egg allergy. Although bird-egg syndrome has been described principally in adults, especially in women, it can also affect children in whom the syndrome presents certain differentiating characteristics in relation to the more common sensitization to egg white. Gastrointestinal and respiratory symptoms are more common than cutaneous symptoms and sensitization to egg yolk is more frequent than that to egg white. In children with allergy to birds and egg, egg allergy is usually more persistent; tolerance is not always achieved and develops later. Sensitization to other aeroallergens is also greater in individuals with allergy to birds and egg. Sensitization to egg sometimes precedes respiratory sensitization to bird proteins, a process known as bird-egg syndrome. By way of example, the case of a child who clinically presented bird egg syndrome is presented at the end of this review. PMID- 12783768 TI - [Effects of childhood nutrition on adult health]. AB - Nutrition and health occupy a long period from the fetal age to adolescence. The interaction between genes and nutrients is one of the most interesting aspects. Health depends on genetic and environmental factors, the most important of which is nutrition. There is a need to define mechanisms and evaluate specific hypotheses in controlled animal experiments and clinical trials in humans. The effects must be permanent, remaining long after the nutritional deficiency or excess has disappeared. This presupposes the existence of critical periods in organ development and biochemical or physiological maturation, during which stimuli leave a permanent imprint. Nutrients can affect the structural development of certain organs during critical periods, both in the fetal and neonatal periods, and can contribute to the development of a different organ structure during these critical periods, leaving permanent effects.The role of folic acid as the prime example of the interaction between genes and nutrients is well known. Folates play an important role in DNA synthesis because of their intervention in deoxyuridine and deoxythymidine methylation and their absence provokes severe disorders in DNA replication and synthesis, which can be precancerous. Periconceptional administration of folic acid is essential to prevent dysraphias (spina bifida). Equally, this nutrient is also important to homocysteine metabolism, which is derived from methionine metabolism and which is an important endothelial toxin. Pediatrics should carefully redefine the criteria for adapting feeding and nutrition not only to childhood health but also to adult well being and disease-free survival. The advent of molecular genetics and the possibility of identifying at-risk individuals and the risks to which they are exposed at an early age, even before birth, provides pediatricians with unique opportunities. However, the growth of knowledge and techniques makes it impossible to keep up to date with all the scientific findings concerning children's health. Nevertheless, as doctors, pediatricians need I would say they are obliged to keep up to date in order to serve as a bridge between science and the practical needs of children and their families. Pediatricians will continue to focus on children's health, although the emphasis will increasingly be placed on prevention rather than cure, and must be ever more aware that the child is the father of the man and that the future is in his hands. PMID- 12783769 TI - Women's networks and the social world of fertility behavior. AB - CONTEXT: Demographic phenomena occur within social contexts and, therefore, should be studied as social processes. However, how to conceptualize and measure the social worlds that individuals inhabit has been the subject of debate. METHODS: Data from a study conducted in Mali in 1996-1997 are used to explore the social networks of Bamanan women and their impact on fertility decisions. Ordinary least-squares and logistic regression techniques are employed to examine the relationship between selected household and social network characteristics and two fertility measures: children ever born and ever-use of contraceptives. RESULTS: Household characteristics do not have a significant effect on either outcome, whereas network attributes do. The more prominently conjugal kin are represented in a network, the fewer children a woman has ever had; however, fertility increases if the husband or unrelated older women are part of the network. Ever-use of contraceptives is elevated if the woman participates in a credit scheme, and rises as the proportion of network members located outside the village increases; it declines sharply as the proportion of network members who are conjugal kin increases, and is significantly elevated if the woman's mother is present. Network effects on fertility are much more pronounced for women aged 30 or older than for younger women, and network effects on contraceptive use are markedly different for younger and older women. CONCLUSIONS: Programs should consider not only women's individual and household characteristics, but also their larger social networks. Additionally, programs should be designed for specific age-groups, given the different network effects on older and younger women. PMID- 12783770 TI - Impact and determinants of sex preference in Nepal. AB - CONTEXT: Gender discrimination and son preference are key demographic features of South Asia and are well documented for India. However, gender bias and sex preference in Nepal have received little attention. METHODS: 1996 Nepal Demographic and Health Survey data on ever-married women aged 15-49 who did not desire any more children were used to investigate levels of gender bias and sex preference. The level of contraceptive use and the total fertility rate in the absence of sex preference were estimated, and logistic regression was performed to analyze the association between socioeconomic and demographic variables and stopping childbearing after the birth of a son. RESULTS: Commonly used indicators of gender bias, such as sex ratio at birth and sex-specific immunization rates, do not suggest a high level of gender discrimination in Nepal. However, sex preference decreases contraceptive use by 24% and increases the total fertility rate by more than 6%. Women's contraceptive use, exposure to the media, parity, last birth interval, educational level and religion are linked to stopping childbearing after the birth of a boy, as is the ethnic makeup of the local area. CONCLUSIONS: The level of sex preference in Nepal is substantial. Sex preference is an important barrier to the increase of contraceptive use and decline of fertility in the country; its impact will be greater as desired family size declines. PMID- 12783771 TI - The link between quality of care and contraceptive use. AB - CONTEXT: Little empirical evidence shows whether quality of family planning care influences continued use of contraception, and if so, to what extent. METHODS: Interviews were conducted in 1997-1998 with 1,728 new family planning users who had sought services at 80 service delivery points in Davao del Norte and Compostela Valley, the Philippines, to assess the quality of care received from family planning services providers. More than 16 months later, 1,460 of the respondents participated in a follow-up survey; the respondents' current contraceptive status was recorded. RESULTS: The quality of care received at the time a woman adopted a contraceptive method influenced her contraceptive use at follow-up, after adjustment for the effects of background characteristics. Furthermore, use increased steadily with quality: The predicted probabilities of contraceptive use were 55% for low-quality care, 62% for medium-quality care and 67% for high-quality care. CONCLUSION: A focus on quality improvement would benefit both programs and users. PMID- 12783772 TI - Knowledge and perception of emergency contraception among female Nigerian undergraduates. AB - CONTEXT: The reproductive health hazards of unintended pregnancies and unsafe abortions are well documented. The potential of emergency contraceptives to prevent unwanted pregnancy in developed countries has been described, but in Nigeria, the awareness about the method is poor and no study has looked at efficacy. METHODS: Between September and October 2001, a randomly selected sample of female undergraduate students at the University of Benin, Nigeria, were surveyed about their demographic information, sexual history and contraceptive use, and their awareness and knowledge of emergency contraception. RESULTS: Of the 880 respondents, 43% were sexually active, 39% had ever practiced contraception and 34% had ever had an induced abortion. Overall, 58% of respondents reported knowing about emergency contraception; sexually active respondents were significantly more likely than those who were not and those who had ever practiced contraception were more likely than those who had not to be aware of emergency contraceptives. However, only 18% of respondents who reported knowing about emergency contraception knew the correct time frame in which emergency contraceptives must be used to be effective. CONCLUSION: There is an urgent need to educate Nigerian young people about emergency contraception, emphasizing available methods and correct timing of use. PMID- 12783773 TI - From home to clinic and family planning to family health: client and community responses to health sector reforms in Bangladesh. PMID- 12783774 TI - Influence of prolonged endurance cycling and recovery diet on intramuscular triglyceride content in trained males. AB - Intramuscular triglycerides (IMTG) are assumed to form an important substrate source during prolonged endurance exercise in trained males. This study investigated the effects of endurance exercise and recovery diet on IMTG content in vastus lateralis muscle. Nine male cyclists were provided with a standardized diet for 3 days, after which they performed a 3-h exercise trial at a 55% maximum workload. Before and immediately after exercise and after 24 and 48 h of recovery, magnetic resonance spectroscopy (MRS) was performed to quantitate IMTG content. Muscle biopsies were taken after 48 h of recovery to determine IMTG content by using quantitative fluorescence microscopy. The entire procedure was performed two times; in one trial, a normal diet containing 39% energy (En%) as fat was provided (NF) and in the other a typical carbohydrate-rich athlete's diet (LF: 24 En% fat) was provided. During exercise, IMTG content decreased by 21.4 +/ 3.1%. During recovery, IMTG content increased significantly in the NF trial only, reaching preexercise levels within 48 h. In accord with MRS, fluorescence microscopy showed significantly higher IMTG content in the NF compared with the LF trial, with differences restricted to the type I muscle fibers (2.1 +/- 0.2 vs. 1.4 +/- 0.2% area lipid staining, respectively). In conclusion, IMTG content in the vastus lateralis muscle declines significantly during prolonged endurance exercise in male cyclists. When a normal diet is used, IMTG contents are subsequently repleted within 48 h of postexercise recovery. In contrast, IMTG repletion is impaired substantially when a typical, carbohydrate-rich athlete's diet is used. Data obtained by quantitative fluorescence microscopy correspond well with MRS results, implying that both are valid methods to quantify IMTG content. PMID- 12783775 TI - Inhibition of Foxo1 function is associated with improved fasting glycemia in diabetic mice. AB - Excessive hepatic glucose production is a contributing factor to fasting hyperglycemia in diabetes. Insulin suppresses hepatic glucose production by inhibiting the expression of two gluconeogenic enzymes, phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G-6-Pase). The forkhead transcription factor Foxo1 has been implicated as a mediator of insulin action in regulating hepatic gluconeogenesis, and a Foxo1 mutant (Foxo1-Delta256), devoid of its carboxyl domain, has been shown to interfere with Foxo1 function and inhibit gluconeogenic gene expression in cultured cells. To study the effect of Foxo1-Delta256 on glucose metabolism in animals, the Foxo1-Delta256 cDNA was delivered to the livers of mice by adenovirus-mediated gene transfer. Hepatic Foxo1-Delta256 production resulted in inhibition of gluconeogenic activity, as evidenced by reduced PEPCK and G-6-Pase expression in the liver. Mice treated with the Foxo1-Delta256 vector exhibited significantly reduced blood glucose levels. In contrast, blood glucose levels in control vector-treated animals remained unchanged, which coincided with the lack of alterations in the expression levels of PEPCK and G-6-Pase. When tested in diabetic db/db mice, hepatic production of Foxo1-Delta256 was shown to reduce fasting hyperglycemia. Furthermore, we showed that hepatic Foxo1 expression was deregulated as a result of insulin resistance in diabetic mice and that Foxo1-Delta256 interfered with Foxo1 function via competitive binding to target promoters. These results demonstrated that functional inhibition of Foxo1, caused by hepatic expression of its mutant, is associated with reduced hepatic gluconeogenic activity and improved fasting glycemia in diabetic mice. PMID- 12783776 TI - Aging is associated with elevated muscle triglyceride content and increased insulin-stimulated fatty acid uptake. AB - The purpose of the present study was to examine the utilization of fatty acids (FA) and muscle substrates by skeletal muscle in young, middle-aged, and old adult rats under hyperglycemic and hyperinsulinemic conditions. Male Fischer 344 x Brown Norway rats aged 5, 15, or 24 mo underwent hindlimb perfusion with a medium of 20 mM glucose, 1 mM palmitate, 1,000 microU/ml insulin, [1 14C]palmitate, and [3-3H]glucose. Glucose uptake and palmitate delivery were similar among age groups. Palmitate uptake and oxidation as well as muscle protein concentration of fatty acid translocase (FAT/CD36) and plasma membrane fatty acid-binding protein (FABPPM) were significantly increased (P < or = 0.05) in 24- vs. 5- and 15-mo-old animals. Compared with 5- and 15-mo-old animals, pre- and postperfusion muscle triglyceride (TG) levels were significantly (P < 0.05) elevated 72-145% in red and 112-129% in white muscles of 24-mo-old animals. Palmitate uptake was associated with total preperfusion TG concentration (r2 = 0.27, P < 0.05) and total TG synthesis rate (r2 = 0.68, P < 0.05). These results indicate that, under insulin-stimulated conditions, FA uptake is significantly increased in old animals, which is associated with increased rates of TG synthesis and may contribute to the accumulation of TG in muscle of old animals. PMID- 12783777 TI - High glucose-mediated effects on endothelial cell proliferation occur via p38 MAP kinase. AB - The mitogen-activated protein (MAP) kinases contribute to altered cell growth and function in a variety of disease states. However, their role in the endothelial complications of diabetes mellitus remains unclear. Human endothelial cells were exposed for 72 h to 5 mM (control) or 25 mM (high) glucose or 5 mM glucose plus 20 mM mannitol (osmotic control). The roles of p38 and p42/44 MAP kinases in the high glucose-induced growth effects were determined by assessment of phosphorylated MAP kinases and their downstream activators by Western blot and by pharmacological inhibition of these MAP kinases. Results were expressed as a percentage (means +/- SE) of control. High glucose increased the activity of total and phosphorylated p38 MAP kinase (P < 0.001) and p42/44 MAP kinase (P < 0.001). Coexposure of p38 MAP kinase blocker with high glucose reversed the antiproliferative but not the hypertrophic effects associated with high-glucose conditions. Transforming growth factor (TGF)-beta1 increased the levels of phosphorylated p38 MAP kinase, and p38 MAP kinase blockade reversed the antiproliferative effects of this cytokine. The high glucose-induced increase in phosphorylated p38 MAP kinase was reversed in the presence of TGF-beta1 neutralizing antibody. Although hyperosmolarity also induced antiproliferation (P < 0.0001) and cell hypertrophy (P < 0.05), there was no change in p38 activity, and therefore inhibition of p38 MAP kinase had no influence on these growth responses. Blockade of p42/44 MAP kinase had no effect on the changes in endothelial cell growth induced by either high glucose or hyperosmolarity. High glucose increased p42/44 and p38 MAP kinase activity in human endothelial cells, but only p38 MAP kinase mediated the antiproliferative growth response through the effects of autocrine TGF-beta1. High glucose-induced endothelial cell hypertrophy was independent of activation of the MAP kinases studied. In addition, these effects were independent of any increase in osmolarity associated with high-glucose exposure. PMID- 12783778 TI - An internal enhancer regulates heme- and cadmium-mediated induction of human heme oxygenase-1. AB - Heme oxygenase-1 (HO-1) catalyzes the rate-limiting step in heme degradation, releasing iron, carbon monoxide, and biliverdin. Induction of HO-1 is an adaptive and beneficial response in renal and nonrenal settings of tissue injury. The purpose of this study was to characterize the regulation of the human HO-1 gene in renal proximal tubule and aortic endothelial cells in response to heme and cadmium. Evaluation of multiple human HO-1 promoter-reporter constructs up to 9.1 kb demonstrated only a partial response to heme and cadmium. In an effort to mimic endogenous stimulus-dependent levels of HO-1 induction, we evaluated the entire 12.5 kb of the human HO-1 gene, including introns and exons, in conjunction with a -4.5-kb human HO-1 promoter and observed significant heme- and cadmium-mediated induction of the reporter gene, suggesting the presence of an internal enhancer. Enhancer function was orientation independent and required a region between -3.5 and -4.5 kb of the human HO-1 promoter. Our studies identified a novel enhancer internal to the human HO-1 gene that, in conjunction with the HO-1 promoter, recapitulates heme- and cadmium-mediated induction of the endogenous HO-1 gene. Elucidation of the molecular regulation of the human HO-1 gene will allow for the development of therapeutic strategies to manipulate HO-1 gene expression in pathological states. PMID- 12783779 TI - Urotensin II is a nitric oxide-dependent vasodilator and natriuretic peptide in the rat kidney. AB - Recent studies have indicated that urotensin II (UII), a cyclic peptide, is vasoactive and may be involved in cardiovascular dysfunctions. It remains unknown, however, whether UII plays a role in the control of renal vascular tone and tubular function. In the present study, a continuous infusion of synthetic human UII (hUII) into the renal artery (RA) in anesthetized rats was found to increase renal blood flow (RBF) and urinary water and sodium excretion (UV and UNaV) in a dose-dependent manner. At a dose of 20 ng. kg-1. min-1, it increased RBF by 20% and UV and UNaV by 94 and 109%, respectively. Nitric oxide (NO) synthase inhibitor NG-nitro-l-arginine methyl ester (l-NAME) completely abolished hUII-induced increases in RBF and water/sodium excretion. In isolated, pressurized, and phenylephrine-precontracted small RA with internal diameter of approximately 200 microm, hUII produced a concentration-dependent vasodilation with a maximal response of 55% at 1.5 microM. l-NAME significantly blocked this hUII-induced vasodilation by 60%. In denuded RA, hUII had neither vasodilator nor vasoconstrictor effect. With the use of 4,5-diaminofluorescein diacetate-based fluorescence imaging analysis of NO levels, hUII (1 microM) was shown to double the NO levels within the endothelium of freshly dissected small RA, and l-NAME blocked this UII-induced production of endothelial NO. These results indicate that UII produces vasodilator and natriuretic effects in the kidney and that UII induced vasodilation is associated with increased endothelial NO in the RA. PMID- 12783781 TI - Novel level of signalling control in the JAK/STAT pathway revealed by in situ visualisation of protein-protein interaction during Drosophila development. AB - It is commonly accepted that activation of most signalling pathways is induced by ligand receptor dimerisation. This belief has been challenged for some vertebrate cytokine receptors of the JAK/STAT pathway. Here we study whether DOME, the Drosophila receptor of the JAK/STAT pathway, can dimerise and if the dimerisation is ligand-dependent. To analyse DOME homo-dimerisation, we have applied a beta gal complementation technique that allows the detection of protein interactions in situ. This technique has been used previously in cell culture but this is the first time that it has been applied to whole embryos. We show that this technique, which we rename betalue-betalau technique, can be used to detect DOME homo-dimerisation in Drosophila developing embryos. Despite DOME being ubiquitously expressed, dimerisation is developmentally regulated. We investigate the state of DOME dimerisation in the presence or absence of ligand and show that DOME dimerisation is not ligand-induced, indicating that ligand independent cytokine receptor dimerisation is a conserved feature across phyla. We have further analysed the functional significance of ligand-independent receptor dimerisation by comparing the effects of ectopic ligand expression in cells in which the receptor is, or is not, dimerised. We show that ligand expression can only activate STAT downstream targets or affect embryo development in cells in which the receptor is dimerised. These results suggest a model in which ligand independent dimerisation of the JAK/STAT receptor confers cells with competence to activate the pathway prior to ligand reception. Thus, competence to induce the JAK/STAT signalling pathway in Drosophila can be regulated by controlling receptor dimerisation prior to ligand binding. These results reveal a novel level of JAK/STAT signalling regulation that could also apply to vertebrates. PMID- 12783782 TI - Six1 is required for the early organogenesis of mammalian kidney. AB - The murine Six gene family, homologous to Drosophila sine oculis (so) which encodes a homeodomain transcription factor, is composed of six members (Six1-6). Among the six members, only the Six2 gene has been previously shown to be expressed early in kidney development, but its function is unknown. We have recently found that the Six1 gene is also expressed in the kidney. In the developing kidney, Six1 is expressed in the uninduced metanephric mesenchyme at E10.5 and in the induced mesenchyme around the ureteric bud at E11.5. At E17.5 to P0, Six1 expression became restricted to a subpopulation of collecting tubule epithelial cells. To study its in vivo function, we have recently generated Six1 mutant mice. Loss of Six1 leads to a failure of ureteric bud invasion into the mesenchyme and subsequent apoptosis of the mesenchyme. These results indicate that Six1 plays an essential role in early kidney development. In Six1(-/-) kidney development, we have found that Pax2, Six2 and Sall1 expression was markedly reduced in the metanephric mesenchyme at E10.5, indicating that Six1 is required for the expression of these genes in the metanephric mesenchyme. In contrast, Eya1 expression was unaffected in Six1(-/-) metanephric mesenchyme at E10.5, indicating that Eya1 may function upstream of Six1. Moreover, our results show that both Eya1 and Six1 expression in the metanephric mesenchyme is preserved in Pax2(-/-) embryos at E10.5, further indicating that Pax2 functions downstream of Eya1 and Six1 in the metanephric mesenchyme. Thus, the epistatic relationship between Pax, Eya and Six genes in the metanephric mesenchyme during early kidney development is distinct from a genetic pathway elucidated in the Drosophila eye imaginal disc. Finally, our results show that Eya1 and Six1 genetically interact during mammalian kidney development, because most compound heterozygous embryos show hypoplastic kidneys. These analyses establish a role for Six1 in the initial inductive step for metanephric development. PMID- 12783783 TI - Loss of Bmp7 and Fgf8 signaling in Hoxa13-mutant mice causes hypospadia. AB - In humans and mice, mutations in Hoxa13 cause malformation of limb and genitourinary (GU) regions. In males, one of the most common GU malformations associated with loss of Hoxa13 function is hypospadia, a condition defined by the poor growth and closure of the urethra and glans penis. By examining early signaling in the developing mouse genital tubercle, we show that Hoxa13 is essential for normal expression of Fgf8 and Bmp7 in the urethral plate epithelium. In Hoxa13(GFP)-mutant mice, hypospadias occur as a result of the combined loss of Fgf8 and Bmp7 expression in the urethral plate epithelium, as well as the ectopic expression of noggin (Nog) in the flanking mesenchyme. In vitro supplementation with Fgf8 restored proliferation in homozygous mutants to wild-type levels, suggesting that Fgf8 is sufficient to direct early proliferation of the developing genital tubercle. However, the closure defects of the distal urethra and glans can be attributed to a loss of apoptosis in the urethra, which is consistent with reduced Bmp7 expression in this region. Mice mutant for Hoxa13 also exhibit changes in androgen receptor expression, providing a developmental link between Hoxa13-associated hypospadias and those produced by antagonists to androgen signaling. Finally, a novel role for Hoxa13 in the vascularization of the glans penis is also identified. PMID- 12783784 TI - Neural crest induction by paraxial mesoderm in Xenopus embryos requires FGF signals. AB - At the border of the neural plate, the induction of the neural crest can be achieved by interactions with the epidermis, or with the underlying mesoderm. Wnt signals are required for the inducing activity of the epidermis in chick and amphibian embryos. Here, we analyze the molecular mechanisms of neural crest induction by the mesoderm in Xenopus embryos. Using a recombination assay, we show that prospective paraxial mesoderm induces a panel of neural crest markers (Slug, FoxD3, Zic5 and Sox9), whereas the future axial mesoderm only induces a subset of these genes. This induction is blocked by a dominant negative (dn) form of FGFR1. However, neither dnFGFR4a nor inhibition of Wnt signaling prevents neural crest induction in this system. Among the FGFs, FGF8 is strongly expressed by the paraxial mesoderm. FGF8 is sufficient to induce the neural crest markers FoxD3, Sox9 and Zic5 transiently in the animal cap assay. In vivo, FGF8 injections also expand the Slug expression domain. This suggests that FGF8 can initiate neural crest formation and cooperates with other DLMZ-derived factors to maintain and complete neural crest induction. In contrast to Wnts, eFGF or bFGF, FGF8 elicits neural crest induction in the absence of mesoderm induction and without a requirement for BMP antagonists. In vivo, it is difficult to dissociate the roles of FGF and WNT factors in mesoderm induction and neural patterning. We show that, in most cases, effects on neural crest formation were parallel to altered mesoderm or neural development. However, neural and neural crest patterning can be dissociated experimentally using different dominant-negative manipulations: while Nfz8 blocks both posterior neural plate formation and neural crest formation, dnFGFR4a blocks neural patterning without blocking neural crest formation. These results suggest that different signal transduction mechanisms may be used in neural crest induction, and anteroposterior neural patterning. PMID- 12783785 TI - Splits ends is a tissue/promoter specific regulator of Wingless signaling. AB - Wingless directs many developmental processes in Drosophila by regulating expression of specific target genes through a conserved signaling pathway. Although many nuclear factors have been implicated in mediating Wingless-induced transcription, the mechanism of how Wingless regulates different targets in different tissues remains poorly understood. We report here that the split ends gene is required for Wingless signaling in the eye, wing and leg imaginal discs. Expression of a dominant-negative version of split ends resulted in more dramatic reductions in Wingless signaling than split ends-null alleles, suggesting that it may have a redundant partner. However, removal of split ends or expression of the dominant-negative had no effect on several Wingless signaling readouts in the embryo. The expression pattern of Split ends cannot explain this tissue-specific requirement, as the protein is predominantly nuclear and present throughout embryogenesis and larval tissues. Consistent with its nuclear location, the split ends dominant-negative acts downstream of Armadillo stabilization. Our data indicate that Split ends is an important positive regulator of Wingless signaling in larval tissues. However, it has no detectable role in the embryonic Wingless pathway, suggesting that it is a tissue or promoter-specific factor. PMID- 12783786 TI - Arabidopsis CROOKED encodes for the smallest subunit of the ARP2/3 complex and controls cell shape by region specific fine F-actin formation. AB - The generation of a specific cell shape requires differential growth, whereby specific regions of the cell expand more relative to others. The Arabidopsis crooked mutant exhibits aberrant cell shapes that develop because of mis-directed expansion, especially during a rapid growth phase. GFP-aided visualization of the F-actin cytoskeleton and the behavior of subcellular organelles in different cell types in crooked and wild-type Arabidopsis revealed that localized expansion is promoted in cellular regions with fine F-actin arrays but is restricted in areas that maintain dense F-actin. This suggested that a spatiotemporal distinction between fine versus dense F-actin in a growing cell could determine the final shape of the cell. CROOKED was molecularly identified as the plant homolog of ARPC5, the smallest sub-unit of the ARP2/3 complex that in other organisms is renowned for its role in creating dendritic arrays of fine F-actin. Rescue of crooked phenotype by the human ortholog provides the first molecular evidence for the presence and functional conservation of the complex in higher plants. Our cell-biological and molecular characterization of CROOKED suggests a general actin-based mechanism for regulating differential growth and generating cell shape diversity. PMID- 12783787 TI - UNC-71, a disintegrin and metalloprotease (ADAM) protein, regulates motor axon guidance and sex myoblast migration in C. elegans. AB - The migration of cells and growth cones is a process that is guided by extracellular cues and requires the controlled remodeling of the extracellular matrix along the migratory path. The ADAM proteins are important regulators of cellular adhesion and recognition because they can combine regulated proteolysis with modulation of cell adhesion. We report that the C. elegans gene unc-71 encodes a unique ADAM with an inactive metalloprotease domain. Loss-of-function mutations in unc-71 cause distinct defects in motor axon guidance and sex myoblast migration. Many unc-71 mutations affect the disintegrin and the cysteine rich domains, supporting a major function of unc-71 in cell adhesion. UNC-71 appears to be expressed in a selected set of cells. Genetic mosaic analysis and tissue-specific expression studies indicate that unc-71 acts in a cell non autonomous manner for both motor axon guidance and sex myoblast migration. Finally, double mutant analysis of unc-71 with other axon guidance signaling molecules suggests that UNC-71 probably functions in a combinatorial manner with integrins and UNC-6/netrin to provide distinct axon guidance cues at specific choice points for motoneurons. PMID- 12783788 TI - Stem cell homeostasis in the Arabidopsis shoot meristem is regulated by intercellular movement of CLAVATA3 and its sequestration by CLAVATA1. AB - Stem cell maintenance in the Arabidopsis shoot meristem is regulated by communication between the apical stem cells and the underlying organizing centre. Expression of the homeobox gene WUSCHEL in the organizing centre induces stem cell identity in the overlying neighbours, which then express the CLAVATA3 gene whose activity in turn restricts the size of the WUSCHEL expression domain. We have analyzed how the stem cells and the organizing centre communicate, by studying the mode of action of CLAVATA3 protein. We provide direct evidence that CLAVATA3 protein functions as a mobile intercellular signal in the shoot apical meristem that spreads laterally from the stem cells and acts both on their lateral neighbours and on the stem cells themselves to repress WUSCHEL transcription. We also show that the spread and range of action of CLAVATA3 can be limited by binding to its receptor CLAVATA1, which offers an explanation for how CLAVATA3 is prevented from entering the organizing centre and repressing WUSCHEL transcription there. This regulated spread of a secreted signalling molecule enables the shoot meristem to permit the onset of cell differentiation in the periphery, but at the same time to maintain a stable niche for its stem cells in the center. PMID- 12783789 TI - Wnt11 and Ret/Gdnf pathways cooperate in regulating ureteric branching during metanephric kidney development. AB - Reciprocal cell-cell interactions between the ureteric epithelium and the metanephric mesenchyme are needed to drive growth and differentiation of the embryonic kidney to completion. Branching morphogenesis of the Wolffian duct derived ureteric bud is integral in the generation of ureteric tips and the elaboration of the collecting duct system. Wnt11, a member of the Wnt superfamily of secreted glycoproteins, which have important regulatory functions during vertebrate embryonic development, is specifically expressed in the tips of the branching ureteric epithelium. In this work, we explore the role of Wnt11 in ureteric branching and use a targeted mutation of the Wnt11 locus as an entrance point into investigating the genetic control of collecting duct morphogenesis. Mutation of the Wnt11 gene results in ureteric branching morphogenesis defects and consequent kidney hypoplasia in newborn mice. Wnt11 functions, in part, by maintaining normal expression levels of the gene encoding glial cell-derived neurotrophic factor (Gdnf). Gdnf encodes a mesenchymally produced ligand for the Ret tyrosine kinase receptor that is crucial for normal ureteric branching. Conversely, Wnt11 expression is reduced in the absence of Ret/Gdnf signaling. Consistent with the idea that reciprocal interaction between Wnt11 and Ret/Gdnf regulates the branching process, Wnt11 and Ret mutations synergistically interact in ureteric branching morphogenesis. Based on these observations, we conclude that Wnt11 and Ret/Gdnf cooperate in a positive autoregulatory feedback loop to coordinate ureteric branching by maintaining an appropriate balance of Wnt11 expressing ureteric epithelium and Gdnf-expressing mesenchyme to ensure continued metanephric development. PMID- 12783790 TI - The T-box-encoding Dorsocross genes function in amnioserosa development and the patterning of the dorsolateral germ band downstream of Dpp. AB - Dpp signals are responsible for establishing a variety of cell identities in dorsal and lateral areas of the early Drosophila embryo, including the extra embryonic amnioserosa as well as different ectodermal and mesodermal cell types. Although we have a reasonably clear picture of how Dpp signaling activity is modulated spatially and temporally during these processes, a better understanding of how these signals are executed requires the identification and characterization of a collection of downstream genes that uniquely respond to these signals. In the present study, we describe three novel genes, Dorsocross1, Dorsocross2 and Dorsocross3, which are expressed downstream of Dpp in the presumptive and definitive amnioserosa, dorsal ectoderm and dorsal mesoderm. We show that these genes are good candidates for being direct targets of the Dpp signaling cascade. Dorsocross expression in the dorsal ectoderm and mesoderm is metameric and requires a combination of Dpp and Wingless signals. In addition, a transverse stripe of expression in dorsoanterior areas of early embryos is independent of Dpp. The Dorsocross genes encode closely related proteins of the T box domain family of transcription factors. All three genes are arranged in a gene cluster, are expressed in identical patterns in embryos, and appear to be genetically redundant. By generating mutants with a loss of all three Dorsocross genes, we demonstrate that Dorsocross gene activity is crucial for the completion of differentiation, cell proliferation arrest, and survival of amnioserosa cells. In addition, we show that the Dorsocross genes are required for normal patterning of the dorsolateral ectoderm and, in particular, the repression of wingless and the ladybird homeobox genes within this area of the germ band. These findings extend our knowledge of the regulatory pathways during amnioserosa development and the patterning of the dorsolateral embryonic germ band in response to Dpp signals. PMID- 12783791 TI - Suppression of macho-1-directed muscle fate by FGF and BMP is required for formation of posterior endoderm in ascidian embryos. AB - Specification of germ layers is a crucial event in early embryogenesis. In embryos of the ascidian, Halocynthia roretzi, endoderm cells originate from two distinct lineages in the vegetal hemisphere. Cell dissociation experiments suggest that cell interactions are required for posterior endoderm formation, which has hitherto been thought to be solely regulated by localized egg cytoplasmic factors. Without cell interaction, every descendant of posterior vegetal blastomeres, including endoderm precursors, assumed muscle fate. Cell interactions are required for suppression of muscle fate and thereby promote endoderm differentiation in the posterior endoderm precursors. The cell interactions take place at the 16- to 32-cell stage. Inhibition of cell signaling by FGF receptor and MEK inhibitor also supported the requirement of cell interactions. Consistently, FGF was a potent signaling molecule, whose signaling is transduced by MEK-MAPK. By contrast, such cell interactions are not required for formation of the anterior endoderm. Our results suggest that another redundant signaling molecule is also involved in the posterior endoderm formation, which is likely to be mediated by BMP. Suppression of the function of macho-1, a muscle determinant in ascidian eggs, by antisense oligonucleotide was enough to allow autonomous endoderm specification. Therefore, the cell interactions induce endoderm formation by suppressing the function of macho-1, which is to promote muscle fate. These findings suggest the presence of novel mechanisms that suppress functions of inappropriately distributed maternal determinants via cell interactions after embryogenesis starts. Such cell interactions would restrict the regions where maternal determinants work, and play a key role in marking precise boundaries between precursor cells of different tissue types. PMID- 12783792 TI - Interactions between the secreted protein Amalgam, its transmembrane receptor Neurotactin and the Abelson tyrosine kinase affect axon pathfinding. AB - Two novel dosage-sensitive modifiers of the Abelson tyrosine kinase (Abl) mutant phenotype have been identified. Amalgam (Ama) is a secreted protein that interacts with the transmembrane protein Neurotactin (Nrt) to promote cell:cell adhesion. We have identified an unusual missense ama allele, ama(M109), which dominantly enhances the Abl mutant phenotype, affecting axon pathfinding. Heterozygous null alleles of ama do not show this dominant enhancement, but animals homozygous mutant for both ama and Abl show abnormal axon outgrowth. Cell culture experiments demonstrate the Ama(M109) mutant protein binds to Nrt, but is defective in mediating Ama/Nrt cell adhesion. Heterozygous null alleles of nrt dominantly enhance the Abl mutant phenotype, also affecting axon pathfinding. Furthermore, we have found that all five mutations originally attributed to disabled are in fact alleles of nrt. These results suggest Ama/Nrt-mediated adhesion may be part of signaling networks involving the Abl tyrosine kinase in the growth cone. PMID- 12783793 TI - Semaphorin3a1 regulates angioblast migration and vascular development in zebrafish embryos. AB - Semaphorins are a large family of secreted and cell surface molecules that guide neural growth cones to their targets during development. Some semaphorins are expressed in cells and tissues beyond the nervous system suggesting the possibility that they function in the development of non-neural tissues as well. In the trunk of zebrafish embryos endothelial precursors (angioblasts) are located ventral and lateral to the somites. The angioblasts migrate medially and dorsally along the medial surface of the somites to form the dorsal aorta just ventral to the notochord. Here we show that in zebrafish Sema3a1 is involved in angioblast migration in vivo. Expression of sema3a1 in somites and neuropilin 1, which encodes for a component of the Sema3a receptor, in angioblasts suggested that Sema3a1 regulates the pathway of the dorsally migrating angioblasts. Antisense knockdown of Sema3a1 inhibited the formation of the dorsal aorta. Induced ubiquitous expression of sema3a1 in hsp70:(gfp)sema3a1(myc) transgenic embryos inhibited migration of angioblasts ventral and lateral to the somites and retarded development of the dorsal aorta, resulting in severely reduced blood circulation. Furthermore, analysis of cells that express angioblast markers following induced expression of sema3a1 or in a mutant that changes the expression of sema3a1 in the somites confirmed these results. These data implicate Sema3a1, a guidance factor for neural growth cones, in the development of the vascular system. PMID- 12783794 TI - SDF-9, a protein tyrosine phosphatase-like molecule, regulates the L3/dauer developmental decision through hormonal signaling in C. elegans. AB - The dauer larva of the nematode Caenorhabditis elegans is a good model system for investigating the regulation of developmental fates by environmental cues. Here we show that SDF-9, a protein tyrosine phosphatase-like molecule, is involved in the regulation of dauer larva formation. The dauer larva of sdf-9 mutants is different from a normal dauer larva but resembles the dauer-like larva of daf-9 and daf-12 dauer-constitutive mutants. Like these mutants, the dauer-constitutive phenotypes of sdf-9 mutants were greatly enhanced by cholesterol deprivation. Epistasis analyses, together with the relationship between sdf-9 mutations and daf-9 expression, suggested that SDF-9 increases the activity of DAF-9 or helps the execution of the DAF-9 function. SDF-9 was expressed in two head cells in which DAF-9 is expressed. By their position and by genetic mosaic experiments, we identified these cells as XXXL/R cells, which are known as embryonic hypodermal cells and whose function at later stages is unknown. Killing of the sdf-9 expressing cells in the wild-type first-stage larva induced formation of the dauer-like larva. Since this study on SDF-9 and former studies on DAF-9 showed that the functions of these proteins are related to those of steroids, XXXL/R cells seem to play a key role in the metabolism or function of a steroid hormone(s) that acts in dauer regulation. PMID- 12783795 TI - An endogenous Su(Hw) insulator separates the yellow gene from the Achaete-scute gene complex in Drosophila. AB - The best characterized chromatin insulator in Drosophila is the Suppressor of Hairy wing binding region contained within the gypsy retrotransposon. Although cellular functions have been suggested, no role has been found yet for the multitude of endogenous Suppressor of Hairy wing binding sites. Here we show that two Suppressor of Hairy wing binding sites in the intergenic region between the yellow gene and the Achaete-scute gene complex form a functional insulator. Genetic analysis shows that at least two proteins, Suppressor of Hairy wing and Modifier of MDG4, required for the activity of this insulator, are involved in the transcriptional regulation of Achaete-scute. PMID- 12783796 TI - Wingless signaling regulates the maintenance of ovarian somatic stem cells in Drosophila. AB - Identifying the signals involved in maintaining stem cells is critical to understanding stem cell biology and to using stem cells in future regenerative medicine. In the Drosophila ovary, Hedgehog is the only known signal for maintaining somatic stem cells (SSCs). Here we report that Wingless (Wg) signaling is also essential for SSC maintenance in the Drosophila ovary. Wg is expressed in terminal filament and cap cells, a few cells away from SSCs. Downregulation of Wg signaling in SSCs through removal of positive regulators of Wg signaling, dishevelled and armadillo, results in rapid SSC loss. Constitutive Wg signaling in SSCs through the removal of its negative regulators, Axin and shaggy, also causes SSC loss. Also, constitutive wg signaling causes over proliferation and abnormal differentiation of somatic follicle cells. This work demonstrates that wg signaling regulates SSC maintenance and that its constitutive signaling influences follicle cell proliferation and differentiation. In mammals, constitutive beta-catenin causes over-proliferation and abnormal differentiation of skin cells, resulting in skin cancer formation. Possibly, mechanisms regulating proliferation and differentiation of epithelial cells, including epithelial stem cells, is conserved from Drosophila to man. PMID- 12783797 TI - Direct and concentration-dependent regulation of the proneural gene Neurogenin2 by Pax6. AB - Expression of the proneural gene Neurogenin2 is controlled by several enhancer elements, with the E1 element active in restricted progenitor domains in the embryonic spinal cord and telencephalon that express the homeodomain protein Pax6. We show that Pax6 function is both required and sufficient to activate this enhancer, and we identify one evolutionary conserved sequence in the E1 element with high similarity to a consensus Pax6 binding site. This conserved sequence binds Pax6 protein with low affinity both in vitro and in vivo, and its disruption results in a severe decrease in E1 activity in the spinal cord and in its abolition in the cerebral cortex. The regulation of Neurogenin2 by Pax6 is thus direct. Pax6 is expressed in concentration gradients in both spinal cord and telencephalon. We demonstrate that the E1 element is only activated by high concentrations of Pax6 protein, and that this requirement explains the restriction of E1 enhancer activity to domains of high Pax6 expression levels in the medioventral spinal cord and lateral cortex. By modifying the E1 enhancer sequence, we also show that the spatial pattern of enhancer activity is determined by the affinity of its binding site for Pax6. Together, these data demonstrate that direct transcriptional regulation accounts for the coordination between mechanisms of patterning and neurogenesis. They also provide evidence that Pax6 expression gradients are involved in establishing borders of gene expression domains in different regions of the nervous system. PMID- 12783798 TI - Mutations in Arabidopsis condensin genes disrupt embryogenesis, meristem organization and segregation of homologous chromosomes during meiosis. AB - Proper chromatin condensation and sister chromatid resolution are essential for the maintenance of chromosomal integrity during cell division, and is in part mediated by a conserved multisubunit apparatus termed the condensin complex. The core subunits of the complex are members of the SMC2 (Structural Maintenance of Chromosomes) and SMC4 gene families. We have cloned an Arabidopsis gene, AtCAP E1, which is a functional ortholog of the yeast SMC2 gene. A second, highly homologous SMC2 gene, AtCAPE-2, was identified by the Arabidopsis genome project. SMC2 gene expression in Arabidopsis was correlated with the mitotic activity of tissues, with high level expression observed in meristematic cells. The two genes are differentially expressed with AtCAP-E1 accounting for more than 85% of the total SMC2 transcript pool. The titan3 mutant is the result of a T-DNA insertion into AtCAP-E1, but other than subtle endosperm defects, titan3 is viable and fecund. We identified a T-DNA insertion mutant of AtCAP-E2, which showed no obvious mutant phenotype, indicating that the two genes are functionally redundant. Genetic crosses were employed to examine the consequences of reduced SMC2 levels. Both male and female gametogenesis were compromised in double mutant spores. Embryo lethality was observed for both double homozygous and AtCAP-E1(-/ ), AtCAP-E2(+/-) plants; arrest occurred at or before the globular stage and was associated with altered planes of cell division in both the suspensor and the embryo. Down regulation of both genes by antisense technology, as well as in AtCAP-E1(+/-), AtCAP-E2(-/-) plants results in meristem disorganization and fasciation. Our data are consistent with the interpretation that threshold levels of SMC2 proteins are required for normal development and that AtCAP-E2 may have a higher affinity for its target than AtCAP-E1. PMID- 12783799 TI - Myf5 expression in somites and limb buds of mouse embryos is controlled by two distinct distal enhancer activities. AB - The initiation of skeletal muscle development in the mouse embryo is strictly associated with the expression of the muscle-specific transcription factor Myf5, the first of four myogenic regulatory factors (MRFs) to be expressed in muscle progenitors, and ablation of the Myf5 gene prevents myogenesis. The complex spatiotemporal expression pattern of Myf5 depends on many discrete regulatory elements that are dispersed over long distances throughout the gene locus. These multiple control modules act differently in the various muscle precursor populations, presumably in response to diverse signals that control myogenesis. A potent enhancer region regulating Myf5 expression in limb muscles and somites has been identified previously at -58/-48 kb upstream of the transcriptional start site (Hadchouel et al., 2000). Here, we focus on the physical and functional dissection of this control region. We demonstrate that a conserved sequence of 270 bp located around -57 kb is required and sufficient to drive Myf5 expression in limbs and to maintain it in somites. A second enhancer nearby is responsible for Myf5 transcription in occipital/cranial somites. This enhancer activity also directs expression accurately to the myotome, preventing ectopic expression in the dermomyotome during the second phase of Myf5 gene activation in somites. Our data suggest that the enhancer identified here collaborates with other somitic enhancers to ensure correct myotomal Myf5 expression. Moreover, it constitutes an important element that mediates somitic expression after the initial and transient Myf5 activation through a previously described sonic hedgehog-dependent early epaxial enhancer. PMID- 12783800 TI - The meiotic protein SWI1 is required for axial element formation and recombination initiation in Arabidopsis. AB - We report the detailed characterization of SWITCH1 (SWI1) an Arabidopsis thaliana protein that has been linked with the establishment of sister chromatid cohesion during meiosis. Using a combination of cytological methods including immunolocalization of meiotic chromosome-associated proteins we show that SWI1 is required for formation of axial elements. Our studies reveal that the swi1-2 mutation prevents the formation of RAD51 foci during meiotic prophase and suppresses the chromosome fragmentation phenotype of the recombination-defective dif1-1 mutant. Together, these data suggest that SWI1 may be required for meiotic recombination initiation. Finally we raised an antibody against SWI1 and showed, by immunolocalization coupled with bromodeoxyuridine incorporation experiments, that SWI1 is expressed exclusively in meiotic G(1) and S phase. Thus, SWI1 appears to be required for early meiotic events that are at the crossroad of sister chromatid cohesion, recombination and axial element formation. The possible inter-relationship between these processes and the function of SWI1 are discussed. PMID- 12783801 TI - lin-35/Rb and ubc-18, an E2 ubiquitin-conjugating enzyme, function redundantly to control pharyngeal morphogenesis in C. elegans. AB - The retinoblastoma gene product has been implicated in the regulation of multiple cellular and developmental processes, including a well-defined role in the control of cell cycle progression. The Caenorhabditis elegans retinoblastoma protein homolog, LIN-35, is also a key regulator of cell cycle entry and, as shown by studies of synthetic multivulval genes, plays an important role in the determination of vulval cell fates. We demonstrate an additional and unexpected function for lin-35 in organ morphogenesis. Using a genetic approach to isolate lin-35 synthetic-lethal mutations, we have identified redundant roles for lin-35 and ubc-18, a gene that encodes an E2 ubiquitin-conjugating enzyme closely related to human UBCH7. lin-35 and ubc-18 cooperate to control one or more steps during pharyngeal morphogenesis. Based on genetic and phenotypic analyses, this role for lin-35 in pharyngeal morphogenesis appears to be distinct from its cell cycle-related functions. lin-35 and ubc-18 may act in concert to regulate the levels of one or more critical targets during C. elegans development. PMID- 12783802 TI - A temperature-sensitive mutation in the nodal-related gene cyclops reveals that the floor plate is induced during gastrulation in zebrafish. AB - The floor plate, a specialized group of cells in the ventral midline of the neural tube of vertebrates, plays crucial roles in patterning the central nervous system. Recent work from zebrafish, chick, chick-quail chimeras and mice to investigate the development of the floor plate have led to several models of floor-plate induction. One model suggests that the floor plate is formed by inductive signalling from the notochord to the overlying neural tube. The induction is thought to be mediated by notochord-derived Sonic hedgehog (Shh), a secreted protein, and requires direct cellular contact between the notochord and the neural tube. Another model proposes a role for the organizer in generating midline precursor cells that produce floor plate cells independent of notochord specification, and proposes that floor plate specification occurs early, during gastrulation. We describe a temperature-sensitive mutation that affects the zebrafish Nodal-related secreted signalling factor, Cyclops, and use it to address the issue of when the floor plate is induced in zebrafish. Zebrafish cyclops regulates the expression of shh in the ventral neural tube. Although null mutations in cyclops result in the lack of the medial floor plate, embryos homozygous for the temperature-sensitive mutation have floor plate cells at the permissive temperature and lack floor plate cells at the restrictive temperature. We use this mutant allele in temperature shift-up and shift-down experiments to answer a central question pertaining to the timing of vertebrate floor plate induction. Abrogation of Cyc/Nodal signalling in the temperature-sensitive mutant embryos at various stages indicates that the floor plate in zebrafish is induced early in development, during gastrulation. In addition, continuous Cyclops signalling is required through gastrulation for a complete ventral neural tube throughout the length of the neuraxis. Finally, by modulation of Nodal signalling levels in mutants and in ectopic overexpression experiments, we show that, similar to the requirements for prechordal plate mesendoderm fates, uninterrupted and high levels of Cyclops signalling are required for induction and specification of a complete ventral neural tube. PMID- 12783805 TI - Advanced membrane technology. Proceedings of a conference sponsored by United Engineering Foundation. Castelvecchio Pascoli, Italy. October 14-19, 2002. PMID- 12783803 TI - Laminin alpha subunits and their role in C. elegans development. AB - Laminins are heterotrimeric (alpha/beta/gamma) glycoproteins that form a major polymer within basement membranes. Different alpha, beta and gamma subunits can assemble into various laminin isoforms that have different, but often overlapping, distributions and functions. In this study, we examine the contributions of the laminin alpha subunits to the development of C. elegans. There are two alpha, one beta and one gamma laminin subunit, suggesting two laminin isoforms that differ by their alpha subunit assemble in C. elegans. We find that near the end of gastrulation and before other basement membrane components are detected, the alpha subunits are secreted between primary tissue layers and become distributed in different patterns to the surfaces of cells. Mutations in either alpha subunit gene cause missing or disrupted extracellular matrix where the protein normally localizes. Cell-cell adhesions are abnormal: in some cases essential cell-cell adhesions are lacking, while in other cases, cells inappropriately adhere to and invade neighboring tissues. Using electron microscopy, we observe adhesion complexes at improper cell surfaces and disoriented cytoskeletal filaments. Cells throughout the animal show defective differentiation, proliferation or migration, suggesting a general disruption of cell-cell signaling. The results suggest a receptor-mediated process localizes each secreted laminin to exposed cell surfaces and that laminin is crucial for organizing extracellular matrix, receptor and intracellular proteins at those surfaces. We propose this supramolecular architecture regulates adhesions and signaling between adjacent tissues. PMID- 12783806 TI - Membrane contactors in the beverage industry for controlling the water gas composition. AB - In the work described here, membrane contactors are used for coupling the removal of species (oxygen and hydrogen sulfide) present in the water with the water carbonation process. We include both experiments and a theoretical study devoted to the analysis of the transport phenomena that occur in the membrane contactor. The main resistance to mass transport was located at the liquid side. Correlations between Sherwood and Reynolds numbers on the shell side that are suitable for the membrane contactor used to carry out our experiments have been determined. In particular, for Re > 1.6, the expression proposed by Yang and Cussler in 1986: Sh = 0.90 Re(0.40) Sc(0.33) describes the behavior of the system; whereas, for Re between 0.03 and 0.3, a new expression is proposed: Sh = 0.435 Re(1.2)Sc(0.33). A comparison with traditional equipment is also furnished. Membrane contactors offer reduced size, CO(2) consumption, and capital costs. PMID- 12783807 TI - Application of hollow fiber membrane contactors for catalyst recovery in the WPO process. AB - In this work the use of a membrane based liquid extraction process for recovery of the homogeneous catalyst employed in the wet peroxide oxidation process (WPO) is studied. In the WPO process the oxidation agent is the hydroxyl radical that is obtained by using a combination of hydrogen peroxide and a mixture of Fe(II), Cu(II), and Mn(II) in aqueous solution. The mixture of metallic cations permits the almost total degradation of the refractory organic compounds, but the use of metallic salts as catalysts induces additional pollution. To recover the homogeneous catalyst of the WPO process by means of non-dispersive solvent extraction (NDSX) two hollow fiber membrane contactors are employed, one for the extraction step and the second for the back-extraction step. From the initial assays, the extractant LIX 622N was selected for Cu(II) recovery and Cyanex 272 for Fe(II) and Mn(II) recovery. Selective separation of Fe(II) and Mn(II) can be obtained by adjusting the pH of the feed aqueous phase. The three metals are stripped using sulfuric acid to give concentrated solutions of CuSO(4), FeSO(4), and MnSO(4) that can be recycled to the formulation of the catalyst solution of the WPO process. A mathematical model has been proposed to describe the recovery of Cu. Two design parameters are required: the membrane mass transport coefficient of the extraction and stripping modules (k(m) = 3.07 x 10(-7) m/sec) and the equilibrium parameter of the extraction reaction (K(Ex) = 0.0832). PMID- 12783808 TI - Membrane contactors for textile wastewater ozonation. AB - This paper deals with the application of a membrane contactor for the ozone treatment of textile wastewater. Ceramic (alpha-Al(2)O(3)) membranes were chosen because of their ozone resistance. A thin metal oxide (TiO(2) and gamma Al(2)O(3)) layer was deposited on the membrane surface to eliminate large defects. Membranes were characterized by bubble pressure and gas permeability tests. Mass transfer coefficients were calculated by using the double-film theory. Decolorization kinetics were studied with model dye solutions. Decolorization experiments with a real exhausted dyebath (untreated and after biological treatment) were also carried out. The potential advantages of membrane contactors for the treatment of these types of effluents are demonstrated. PMID- 12783809 TI - Closing pulp and paper mill water circuits with membrane filtration. AB - In this study membrane filtration, ultrafiltration, and nanofiltration alone and as part of hybrid processes are considered as means to purify pulp and paper mill process waters suitable for reuse. Thermophilic aerobic biological treatment, pH adjustment, flocculation, and ozonation were tested as pretreatment methods on pilot or on laboratory scale. The aim was to increase flux and reduce fouling by various pretreatment steps and, thus, increase the competitiveness of the membrane process. The results were also evaluated by comparing the benefits obtained against the costs. It was discovered that benefits could be obtained with all the pretreatments tried. Thermophilic aerobic biology assisted in the removal of organic material and increased flux significantly, but the costs were the highest. The most cost-effective processes, however, seem to be pH-adjusted nanofiltration and flocculation nanofiltration hybrid processes, which is understandable because of their significantly lower investment costs compared to, for example, those of biological process. The pH adjustment increased the electrostatic repulsion between negatively charged solutes and membrane, thereby increasing the flux. Flocculation removed the foulants effectively from the feed and it both increased flux and reduced fouling. Yet, many noteworthy benefits were obtained also with ultrafiltration and ozonation. All of the hybrid processes tested could be applied at various points of the water circuit of an integrated pulp and paper mill for purification purposes. The eventual superiority and cost-effectiveness of the applied process remains to be determined case by case. PMID- 12783810 TI - Membrane technologies applied to textile wastewater treatment. AB - This paper describes the experimental results of a pilot-scale application of membrane technologies to textile wastewater advanced treatment, downstream of a biological activated sludge process, aimed at water reuse in textile technology processes. The chosen approach consisted of sand filtration as a pretreatment, a microfiltration (MF) or ultrafiltration (UF) membrane process, and a final separation treatment performed by means of a nanofiltration (NF) or a reverse osmosis (RO) membrane. An experimental study to compare spiral wound membranes, operating under pressure, to flat membranes, operating under vacuum was conducted. The technical results and a preliminary economic analysis indicate the possibility of technological transfer of the membrane technologies to an industrial scale for textile wastewater reclamation. PMID- 12783811 TI - Integrating photocatalysis and membrane technologies for water treatment. AB - Removal of organic contaminants in water may sometimes be more easily achieved with a pair rather than with a single unit operation. We explore here the combination of an ambient temperature, chemical oxidation photocatalysis (PC) process with physical separation via a membrane operation, microfiltration (MF), ultrafiltration (UF), or reverse osmosis (RO) using both a conceptual statement and a literature review format. Four configurations are noted and discussed here: (1) PC + MF for catalyst slurry recycle, (2) PC + UF for catalyst slurry and (polymer) reactant recycle, (3) immobilized PC and UF/RO for reactant recycle, and (4) immobilized PC on UF/RO membrane for membrane self-cleaning. Although the literature review is encouraging with respect to plausibility, the paucity of examples indicates a need for substantial effort to fully exploit the suggested possibilities for process development. PMID- 12783812 TI - Water quality monitoring in membrane filtration systems. AB - We report on an experimental study of UF membrane fouling by colloidal particles. Deposition colloidal particles during membrane filtration causes a decline in permeate flux. Membrane flux is monitored on a laboratory scale, crossflow employing UF membranes. The existing modified fouling index (MFI) uses a microfilter membrane as a quick test of feed water quality. The MFI is based on cake filtration, and thus, a model can be developed for flux decline predication. However, this MFI is not sensitive to the presence of smaller particles. Therefore, more recently MFI using ultrafiltration membranes (MFI-UF) was developed. This research investigates various critical aspects of the MFI-UF test for use as a water quality indicator; stability of the MFI-UF over time, linearity of the index with particulate concentration, and reproducibility (1) of the test (reusability of a UF module) and (2) module manufacture. Pressure dependence of the MFI-UF was also examined. The aforementioned criteria were examined using a polyacrylonitrile module with 13,000 molecular weight cutoff for low fouling (tap and process water). The MFI-UF was stable over time and directly related to colloidal concentration. The MFI-UF test was reproducible for one module with repeated testing; reproducible module manufacture was found for 80% of the test modules. PMID- 12783813 TI - Removal and recovery of metals and other materials by supported liquid membranes with strip dispersion. AB - This paper reviews recent advances in supported liquid membranes (SLMs) with strip dispersion for removal and recovery of metals including chromium, copper, zinc, and strontium; it also discusses potential applications of SLMs for removal and recovery of other materials, including cobalt and penicillin G. The technology for chromium that we developed, not only removes the Cr(VI) from about 100-1,000 ppm to less than 0.05 ppm in the treated effluent allowable for discharge or recycle, but also recovers the chromium product at a high concentration of about 20% Cr(VI) (62.3% Na(2)CrO(4)) suitable for resale or reuse. In other words, we have achieved the goals of zero discharge and no sludge. The stability of the SLM is ensured by a modified SLM with strip dispersion, where the aqueous strip solution is dispersed in the organic membrane solution in a mixer. The strip dispersion formed is circulated from the mixer to the membrane module to provide a constant supply of the organic solution to the membrane pores. The copper SLM system that we have identified, not only removed the copper from 150 ppm in the inlet feed to less than 0.15 ppm in the treated feed, but also recovered the copper at a high concentration of greater than 10,000 ppm in the strip solution. For the zinc SLM system identified, zinc at an inlet feed concentration of 550 ppm was removed to less than 0.3 ppm in the treated feed, whereas a high zinc concentration of more than 17,000 ppm was recovered in the strip solution. For strontium removal, we synthesized a family of new extractants, alkyl phenylphosphonic acids. The SLM removed radioactive (90)Sr to the target of 8 pCi/L or lower from feed solutions of 300-1,000 pCi/L. The SLM removed cobalt from about 525 ppm to 0.7 ppm in the treated feed solution, concentrating it to at least 30,000 ppm in the aqueous strip solution. Concerning penicillin G recovery, the SLM removed penicillin G from a feed of 8,840 ppm and concentrated it to a high concentration of 41,011 ppm in the aqueous strip solution with a high recovery of about 93%. PMID- 12783814 TI - Membrane separation in green chemical processing: solvent nanofiltration in liquid phase organic synthesis reactions. AB - This paper describes ideas together with preliminary experimental results for applying solvent nanofiltration to liquid phase organic synthesis reactions. Membranes for organic solvent nanofiltration have only recently (during the 1990s) become available and, to date, have been applied primarily to food processing (vegetable oil processing, in particular) and refinery processes. Applications to organic synthesis, even at a laboratory feasibility level, are few. However, these membranes have great potential to improve the environmental performance of many liquid phase synthesis reactions by reducing the need for complex solvent handling operations. Examples that are shown to be feasible are solvent exchanges, where it is desired to swap a high molecular weight molecule from one solvent to another between separate stages in a complex synthesis, and recycle and reuse of homogeneous catalysts. In solvent exchanges, nanofiltration is shown to provide a fast and effective means of swapping from a high boiling point solvent to a solvent with a lower boiling point-this is a difficult operation by means of distillation. Solvent nanofiltration is shown to be able to separate two distinct types of homogeneous catalysts, phase transfer catalysts and organometallic catalysts, from their respective reaction products. In both cases the application of organic solvent nanofiltration allows several reuses of the same catalyst. Catalyst stability is shown to be an essential requirement for this technique to be effective. Finally, we present a discussion of scale-up aspects including membrane flux and process economics. PMID- 12783815 TI - Computer-aided simulation and design of nanofiltration processes. AB - The modelling of membrane filtration processes is often performed by applying black-box models or short-cut methods, because of the complexity of the molecular interactions on and inside the membrane. The assumptions made for short-cut methods can be applied with accuracy to reverse osmosis processes, whereas the simulation of nanofiltration can lead to unreliable results that sometimes deviate from real conditions to a great extent. A steady-state process simulation, NF-PROJECT, based on input information from membrane characterization, was developed (isothermal operation). The individual separation characteristics of each membrane element are calculated in an iterative sequence, illustrating the successive reduction in permeability and rejection between the elements arranged inside the pressure vessel. The simulation provides information on the increasing feed concentration and osmotic pressure, the hydraulic pressure loss, the deterioration of the flow conditions in the vessel, and the joint performance of the membrane elements to be analyzed. Taking an example from a practical application, a two-stage nanofiltration pilot plant was simulated, the results of which are presented in this article. Examples of optimization potentials are illustrated for the target criteria of economic efficiency (specific energy costs), permeate quality, and flow. PMID- 12783816 TI - Advances in solvent-resistant nanofiltration membranes: experimental observations and applications. AB - Nanofiltration (NF) and reverse osmosis (RO) are well-established membrane technologies for applications involving aqueous streams. The principles of NF transport (diffusion, convection, and Donnan exclusion) are effectively used to develop novel membrane materials and applications in aqueous medium. Use of NF in a non-aqueous medium holds strong potential for the food, refining, and pharmaceutical industries because of the low energy costs involved with such membrane processes. Further understanding and development of solvent-resistant NF membranes provides opportunities for various hybrid processing ranging from reactor-membrane to distillation-membrane combinations. This paper provides a comprehensive overview of literature results and our own work in the area of non aqueous systems. For solvent-based systems, potential membrane swelling and solvent-solute coupling needs to be considered for membrane design and transport theories. A simplified transport theory for pure solvents has been developed using solvent (molar volume, viscosity) and membrane properties (membrane surface energy). This model and has been verified with literature data for both hydrophilic and hydrophobic membranes. Membrane characterization and preconditioning aspects need to be given serious consideration for evaluating membrane performance. In addition to permeability and separation results, some novel applications of NF in non-aqueous solvents are included in this paper. PMID- 12783817 TI - Characteristics and application of ceramic nanofiltration membranes. AB - In this paper we report on the characteristic and filtration behavior of a newly developed ceramic nanofiltration membrane and compare it with other commercial ceramic nanofiltration membranes currently available. It is shown that it is possible to produce a ceramic membrane with separation properties in the nanofiltration range and with permeability rates that are clearly superior to those of polymer nanofiltration membranes. The ceramic membrane was used in tests involving the treatment of textile wastewater, alkaline solutions from bottle washing machines, and pickling bath solutions. PMID- 12783818 TI - Nanocomposite lithium ion conducting membranes. AB - This review describes the properties and characteristics of a class of membranes formed by blends of a lithium salt, LiX, where X is preferably a large soft anion, such as ClO(4) or N(CF(3)SO(2))(2), and a high molecular weight polymer containing Li(+)-coordinating group, such as polyethylene oxide (PEO) with the dispersion of selected ceramic powders, such as TiO(2), Al(2)O(3), and SiO(2), at the nanoscale particle size. These nanocomposite membranes behave as lithium polymer electrolytes, that is, they exhibit a high lithium ion conductivity. Because of this property, the PEO-LiX nanocomposite electrolytes may find an important application as separators in advanced, rechargeable lithium polymer batteries. PMID- 12783820 TI - High performance perfluoropolymer films and membranes. AB - Membrane processes are receiving increasing attention in the scientific community and in industry because in many cases they offer a favorable alternative to processes that are not easy to achieve by conventional routes. In this context, membranes made with perfluorinated polymers are of particular interest because of the unique features demonstrated by these materials. Both highly hydrophobic and hydrophilic membranes have been developed from appropriate perfluoropolymers that were, in turn, obtained by copolymerizing TFE with special monomers available on an industrial scale. Highly hydrophobic membranes obtained from the glassy copolymers of TFE and 2,2,4-trifluoro-5 trifluoromethoxy-1,3 dioxole (Hyflon AD) exhibit properties that make them particularly well suited for use in optical applications, in the field of gas separation, and in gas-liquid contactors. Conditions for preparing membranes that are adequate for use in various applications are exemplified. Hydrophylic highly conductive proton exchange membranes obtained from the copolymer of TFE and a short-side-chain (SSC) perfluorosulfonylfluoridevinylether (Hyflon Ion) find interesting application in the field of fuel cells, especially in view of the current tendency to move to high temperature operation. The advantages offered by these hydrophobic and hydrophylic perfluorinated materials for use in membrane technology are discussed. Comparison of membrane properties and performance is made with other membranes available on the market. PMID- 12783819 TI - Preparation of nano-structured polymeric proton conducting membranes for use in fuel cells. AB - We briefly discuss the state of the art of polymer electrolyte membrane fuel cells and suggest that the main obstacles to the commercial development of these fuel cells are essentially the high costs and poor characteristics of present proton conducting membranes. A strategy for the preparation of improved nanocomposite membranes based on the introduction of proton conducting lamell? in the polymeric matrix of present ionomeric membranes is then discussed. Due to their high proton conductivity (in some cases even higher than 10(-1) S cm(-1)), tailor made lamellae obtained by exfoliation of superacid metal (IV) phosphonates are particularly suitable for the preparation of these hybrid membranes. The expected positive influence of the dispersed lamellae on important properties of proton conducting membranes, such as swelling, mechanical resistance, proton transport, and diffusion of methanol, are also discussed. The methods used to obtain good lamellar dispersions into ionomeric polymers and the preparation and main characteristics of some hybrid membranes are also briefly described. The presence of nanoparticles of metal phosphonates in the electrodic interfaces Nafion/Pt already considerably improves the electrochemical characteristics of fuel cells in the temperature range 80-130 degrees C. The increased working temperature of the fuel cell considerably reduces CO poisoning of the platinum electrodes and allows better control of the cooling system, thus overcoming important obstacles to the development of medium temperature PEM fuel cells. PMID- 12783821 TI - Novel charge-mosaic membranes. AB - Novel charge-mosaic composite membranes composed of cation and anion permeable domains were prepared. Poly(4-methylstyrene-isoprene-styrene-isoprene-4 methylstyrene) pentablock copolymer (MsISIMs), poly(isoprene-4-methylstyrene) diblock copolymer (IMs), and poly(isoprene-styrene-isoprene) triblock copolymer (ISI), used as the precursor of the charge-mosaic membranes, were polymerized from styrene (S), isoprene (I), and 4-methylstyrene (Ms). MsISIMs and a blend of ISI and IMs were then dissolved in benzene and cast on a polypropylene microporous supporting membrane. After chemical modification, the MsISIMs and IMs/ISI charge-mosaic composite membranes were obtained. Our experimental results show that the IMs/ISI charge-mosaic composite membrane is similar in membrane structure and properties to the MsISIMs charge-mosaic composite membrane. PMID- 12783822 TI - Material transport through charged mosaic membrane. AB - Several characteristics of a charged mosaic membrane with parallel array of negative and positive charges were investigated by using transport studies and the related analysis. From an analysis of the volume flux and salt flux based on irreversible thermodynamics, preferential salt transport across the charged mosaic membrane was clearly demonstrated. Additionally, transport properties of amino acids and sucrose through the charged mosaic membrane were estimated, relatively, on the basis of KCl transport. As a result, amino acid transport depends largely on the charged states and molecular weight; however, for sucrose transport, non-electrolyte was rejected under all experimental conditions. PMID- 12783823 TI - Effect of compatibility of PVC/P2 alloy system on membrane structure and performance. AB - The effects of the secondary polymer component (P(2)) on poly(vinyl chloride) (PVC)/P(2) alloy membrane structure and performance were systematically investigated. A series of P(2) with varying compatibility with PVC used in this study included vinyl chloride-vinyl acetate copolymer (VC-co-VAc); copolymer of vinyl chloride, vinyl acetate-maleic anhydride copolymer (VC-co-VAc-co-MAL); isobutylene-maleic anhydride copolymer (IB-co-MAL); poly(methyl methacrylate) (PMMA); poly(vinylidene dichloride) (PVDC); and styrene-acrylonitrile copolymer (SAN). Alloy membranes were prepared by means of solution blending-phase inversion technique. On the basis of our experimental results, the compatibility of PVC/P(2) was proved to be the most critical factor affecting the alloy membrane structure and performance. Systems with good compatibility, such as PVC/VC-co-VAc, are more suitable for preparing membranes with small pore size; whereas systems with partial compatibility, such as PVC/PMMA, are more favored for the formation of large-pore membranes. PMID- 12783824 TI - Stable liquid membranes: recent developments and future directions. AB - Immobilized liquid membranes offer the advantages of high selectivity and species flux in separation of species from either gaseous or liquid streams. Despite these advantages, they are not yet commercially viable because of their instabilities and limited lifetime. This paper presents recent advances made in developing stable immobilized liquid membranes for gas separation with high fluxes and selectivities for the species of interest. Some of the recent approaches being studied for development of stable liquid membranes for liquid separation are also discussed. PMID- 12783826 TI - Natural gas cleanup by means of membranes. AB - This paper deals with the use of membranes for hydrocarbon dewpointing and dehydration of natural gas. Based on experience gained from membrane applications in separating organic vapors from off-gas and process streams, as well as the dehydration of compressed air, membranes have been developed and tested for use in high pressure applications. Membranes and membrane modules have been modified to withstand the high operating pressure. Calculation programs were developed to understand the separation performance and to provide the necessary information for optimizing membrane design. A real challenge was the introduction of the vacuum mode dehydration operation in order to achieve the highest possible dewpoint reduction with minimum methane loss. PMID- 12783825 TI - The use of conducting polymers in membrane-based separations: a review and recent developments. AB - As a material family, pi-conjugated polymers (also known as intrinsically conductive polymers) elicit the possibility of both exploiting the chemical and physical attributes of the polymer for membrane-based separations and incorporating its electronic and electrochemical properties to enhance the separation figures-of-merit. This review article, although by no means comprehensive, provides a current snapshot of the investigations from many research laboratories in the use of conducting polymers for membrane-based separations. The review focuses primarily on polyaniline, polypyrrole, and substituted-polythiophene and includes applications in gas separations, liquid (and/or vapor) separations, and ion separations. Additionally, we discuss the broad challenges and accomplishments in membrane formation from conducting polymers. PMID- 12783827 TI - Polysulfone hollow fiber gas separation membranes filled with submicron particles. AB - Three different fillers, carbon black (CB), vapor grown carbon fibers (VGCF), and TiO(2), were incorporated into polysulfone spinning solutions with the intention of producing highly selective membranes with enhanced mechanical strength. The effect of filler presence on gas permeation characteristics, mechanical strength (bursting pressure), and morphology was investigated and compared to unfilled membranes. As well as studying filler types, the influence of CB filler concentration on membrane performance was also examined. For all filler types (at a concentration of 5%w/w), the pressure-normalized flux of O(2), N(2), and CH(4) was greater in the composite than in the unfilled membranes. The CO(2) pressure normalized flux was only greater in the TiO(2) composite membranes. For CB and VGCF, the CO(2) pressure-normalized flux was reduced compared with unfilled membranes. Three CB concentrations were investigated (2, 5, and 10%w/w). For O(2), N(2), and CH(4), pressure-normalized flux peaked at 5%w/w CB. CO(2) exhibited the opposite trend, showing a minimum pressure-normalized flux at 5%w/w. Considering O(2)/N(2) and CO(2)/CH(4) gas pairs and the various filled membrane categories, only the O(2)/N(2) selectivity of the 2%w/w CB filled membranes was higher than that of the unfilled fibers-all other selectivities were lower. In terms of CB concentration, selectivity was a minimum at the intermediate concentration of 5%w/w. All the filled membrane types exhibited greater mechanical strength (bursting pressure) than unfilled fibers apart from the 5%w/w VGCF composites. The 2%w/w CB composites were the strongest. Electron microscopy showed no visible differences in general morphology between the various filled and unfilled membranes. PMID- 12783828 TI - Carbon molecular sieve membranes: a promising alternative for selected industrial applications. AB - Carbon molecular sieve (CMS) membranes (hollow fibers) have been studied for application as possible separation units for selected industrial gas streams. Gas streams at petrochemical plants (polypropene and polyethene) and upgrading of biogas to fuel specifications have been in focus. Gases present in biogas (N(2), CO(2), H(2)O(vap), and CH(4)) and gas streams at polyolefin plants (C(2)H(4), C(3)H(6), and C(3)H(8)) have been measured; both as pure gases and in mixtures. Aging of the CMS-membranes as a function of humidity and pore blocking is discussed; likewise, possible regeneration methods when flux decrease is experienced. Transport mechanisms depending on pore size and molecular properties are also discussed. Excellent separation properties were documented for these applications, but also the need for frequent regeneration of the membrane in order to maintain permeability flux. The mixed gas experiments documented clearly the need for careful pore tailoring in order to optimize selectivity when the membranes were used for alkane-alkene separation. PMID- 12783829 TI - Thin composite palladium and palladium/alloy membranes for hydrogen separation. AB - Dense composite Pd and Pd/alloy membranes are currently being extensively investigated. The synthesis and characterization of these membranes, with a special emphasis on Pd/alloy membranes, are reviewed in this paper. Experimental results on Pd/Cu membranes supported on porous stainless steel exhibited good thermal stability and reasonable hydrogen flux. Furthermore, optical micrographs showed the formation of the dense palladium layer was unaffected by the topological features of the porous stainless steel, although the surface of the support directs the topology of the final Pd layer. PMID- 12783830 TI - Mixed matrix membrane development. AB - Two types of mixed matrix membranes were developed by UOP in the late 1980s. The first type includes adsorbent polymers, such as silicalite-cellulose acetate (CA), NaX-CA, and AgX-CA mixed matrix membranes. The silicalite-CA has a CO(2)/H(2) selectivity of 5.15 +/- 2.2. In contrast, the CA membrane has a CO(2)/H(2) selectivity of 0.77 +/- 0.06. The second type of mixed matrix membrane is PEG-silicone rubber. The PEG-silicone rubber mixed matrix membrane has high selectivity for polar gases, such as SO(2), NH(3), and H(2)S. PMID- 12783831 TI - Nonlinear parameter estimation for solution-diffusion models of membrane pervaporation. AB - An optimization-based procedure for estimating unknown parameters in solution diffusion models of membrane pervaporation is presented. Permeation of two components through a polymer membrane is described by distinct solution and diffusion models. The solution model is based on a modified form of Flory-Huggins theory that accounts for interactions between the two penetrants. The diffusion model is derived from Fick's law, where the diffusion coefficients are allowed to depend on the local concentration of each component in the membrane. A phenomenologic relation is used to account for the effect of temperature on the component fluxes. The solution and diffusion models, as well as the temperature flux relation, contain parameters that are not directly measurable. It is shown that these parameters can be estimated effectively from sorption and flux data by the solution of suitably formulated nonlinear optimization problems. The separation of styrene and ethylbenzene with a polyurethane membrane is used to illustrate the parameter estimation procedure. PMID- 12783832 TI - Poly(vinyl alcohol)-based polyelectrolyte pervaporation membranes. AB - By modifying poly(vinyl alcohol) (PVA), phosphatic anionic PVA (P-PVA) and quaternary ammonium cationic PVA (C-PVA) with various degrees of substitution (D.S.) were synthesized. The effects of synthesis conditions on the degree of substitution were studied. With these two kinds of materials, polyelectrolyte complexes were formed and their solubilities in water were studied. Pervaporation composite membranes were prepared from P-PVA, C-PVA, and their polyelectrolyte complexes. Some of the membranes showed good separation performance. The polyelectrolyte complex membrane prepared by mixing P-PVA (D.S. 2.3%) and C-PVA (D.S. 2.9%) with weight ratio of 1/1, showed a permeation rate of 378 g/m(2)h and separation factor of 2,250 for dehydration of ethanol/water mixture (ethanol 95.4 wt%) at a feed temperature of 75 degrees C. Some factors that influenced pervaporation performance included type of polyelectrolyte, degree of substitution, ratio of polyanion and polycation, feed temperature, feed concentration, and elapsed time. Solvent resistance of pervaporation composite membranes was also evaluated. PMID- 12783833 TI - Hydrophobic pervaporation: toward a shortcut method for the pervaporation decanter system. AB - Pervaporation is a relatively new technology. Although hydrophilic pervaporation has become established, hydrophobic pervaporation for recovery of organics from water has not been a commercial success. Technologic reasons for this are suggested. However, as the pressure to include waste minimization and to recycle, as well as pollution prevention, increases, there will be opportunity for the development of new wastewater treatment processes. This may lead to hybrid processes, including a coupling of pervaporation with conventional technology. The hybrid process examined herein is a simple pervaporation-decanter system that is applicable to organics with limited solubility in water. In this system, the PV unit produces permeate that after condensation gives two liquid phases. The organic phase is relative pure and concentrated (and available for reuse), whereas the aqueous phase can be recycled into the PV feed stream. For a given feed concentration and water purity target, there is a minimum membrane selectivity (alpha(min)) that yields a two-phase condensate. If the membrane has a selectivity that is just greater than the minimum, the recycle rate of the aqueous phase relative to the feed rate will be very large and likewise the membrane area. Also, for a membrane with known organic permeability, one can define a theoretical A(min), the minimum membrane area required when the separation factor is so high that the aqueous phase is negligible. For membranes with alpha > alpha(min), values of the required membrane area, A, have been obtained for various representative duties, and a correlation between alpha/alpha(min) and A/A(min) has been obtained (compare with the Gilliland correlation used in distillation). An approximate relationship is (A/A(min) (- 1))(alpha/alpha(min) (- 1)) = 1. Since alpha(min) and A(min) can be calculated readily, this relationship is a shortcut tool that permits estimation of A for any alpha and any duty. Finally, membrane areas for the above hybrid system and those for PV alone are compared. PMID- 12783834 TI - Membrane bioreactors for treating waste streams. AB - Membrane bioreactors (MBRs) have a number of advantages for treating wastewater containing large quantities of BOD. This paper reviews the inherent advantages of an MBR, which include high potential biomass loadings, lower sludge yields, and retention of specialized organisms that may not settle well in clarifiers. A major problem in effluent treatment occurs when mixed inorganic and organic wastes occur with high concentrations of pollutants. Inorganics that might cause extremes of pH and/or salinity will inhibit microbial growth and only specialized organisms can survive under these conditions. Refractory organics are only biodegraded with difficulty by specialized organisms, which usually do not resist the extreme inorganic environments. The use of membrane bioreactors to help separate the micro-organisms from the inorganic compounds, yet permit the organics to permeate, has been developed in two different designs that are outlined in this paper. The use of membrane contactors in a multimembrane stripping system to treat acidic chlorinated wastes is proposed and discussed. PMID- 12783835 TI - Why and how membrane bioreactors with unsteady filtration conditions can improve the efficiency of biological processes. AB - A membrane bioreactor (MBR), an association of a bioreactor with a crossflow filtration unit, enables continuous processes with total cell retention within the reactor to be realized. Provided that high dilution rates can be applied and that inhibition processes are avoided, very high biomass concentrations can be reached, thereby improving the volumetric productivities. These membrane bioreactors have been successfully applied to various microbial bioconversion, such as alcoholic fermentation, solvents, organic acid production, starters, and wastewater treatment. On the basis of the biological reaction characteristics and bibliographic results, the potentialities and bottlenecks of this methodology are discussed. Depending on the application, it is shown how the performance of the membrane bioreactor can be enhanced by acting either on the biological reaction achievement, by controlling the balance between cell growth and death, or on the dilution rate, by increasing the permeate flux through the filtration unit. This discussion is based on results obtained in specific biological treatments applied to polluted liquid and gas. PMID- 12783836 TI - Effect of immobilization site and membrane materials on multiphasic enantiocatalytic enzyme membrane reactors. AB - In the experimental work reported here the optical resolution of racemic naproxen methyl ester with crude lipase immobilized in a membrane reactor was studied. The multiphasic enantiocatalytic enzyme membrane reactor consisted of an organic phase that dissolved the naproxen methyl ester, a lipase-loaded membrane, and an aqueous phase that extracted the reaction product. Lipase preferentially converted the (S)-naproxen methyl ester to (S)-naproxen acid that was simultaneously separated by a membrane. The effect of the immobilization site and membrane type on the performance of the enzyme membrane reactor was studied. Capillary polyamide membrane with 10 kDa nominal molecular weight cutoff (NMWCO) and polysulfone membrane with 30 kDa NMWCO were applied, with lipase loaded in the sponge layer or on a membrane thin layer. With various immobilization sites and membrane types, the enzyme membrane reactors showed different productivity and enantioselectivity resulting from the varying amounts of immobilized enzyme and varying microenvironment of the hydrolysis reaction. Higher amounts of immobilized enzyme led to increased productivity and generally higher enantioslectivity of the membrane reactor. It seems that the location of organic/aqueous interface on membrane, which plays an important role in a multiphasic enzyme membrane reactor, was influenced by the immobilization site and membrane type and this affected the productivity and enantioselectivity. As much as 90% enantioexcess was obtained with a polyamide membrane, and lower values with a polysulfone membrane. A sponge layer of polyamide membrane is the preferred immobilization site for its higher productivity than a thin membrane layer, and for its higher enantioselectivity than a polysulfone membrane. In comparison with the unstable hydrolysis activity of free lipase in stirred tank reactor, a stable lipase activity can be obtained with the lipase-immobilized membrane reactor, irrespective of whether a polyamide or polysulfone membrane is used. PMID- 12783837 TI - CO2 capture by means of an enzyme-based reactor. AB - We report a means for efficient and selective extraction of carbon dioxide (CO(2)) at low to medium concentration from mixed gas streams. CO(2) capture was accomplished by use of a novel enzyme-based, facilitated transport contained liquid membrane (EBCLM) reactor. The parametric studies we report explore both structural and operational parameters of this design. The structural parameters include carbonic anhydrase (CA) concentration, buffer concentration and pH, and liquid membrane thickness. The operational parameters are temperature, humidity of the inlet gas stream, and CO(2) concentration in the feed stream. The data show that this system effectively captures CO(2) over the range 400 ppm to at least 100,000 ppm, at or around ambient temperature and pressure. In a single pass across this homogeneous catalyst design, given a feed of 0.1% CO(2), the selectivity of CO(2) versus N(2) is 1,090 : 1 and CO(2) versus O(2) is 790 :1. CO(2) permeance is 4.71 x 10(-8) molm(-2) Pa(-1) sec(-1). The CLM design results in a system that is very stable even in the presence of dry feed and sweep gases. PMID- 12783838 TI - Affinity membranes as a tool for life science applications. AB - As a matrix for affinity membrane technology we chose a flat-sheet microfiltration membrane based on polypropylene. Using photopolymerization to graft epoxy groups onto the pore surface, we worked with glycidylmethacrylate as a monomer. We developed optimized, efficient, and mild UV irradiation conditions for the two-step photografting process practically preserving the given pore structure of the base membrane. A grafting degree of up to 1.2 mg/cm(2) per surface area of the membrane was obtained. The poly-propylene membrane surface became significantly more hydrophilic. Introduction of epoxy groups allowed a stable covalent immobilization of the protein streptavidin serving as receptor for affinity ligand binding. A relatively high streptavidin immobilization capacity of about 65 micro g/cm(2) per surface area of the membrane was obtained. Apparently, only about two of the binding sites of the immobilized streptavidin were available for biotin recognition. We also found that the oriented immobilization of biotinylated alkaline phosphatase onto the surface via a streptavidin bridge increased the specific enzymatic activity about sixfold compared with random immobilization of this enzyme. PMID- 12783839 TI - Enzyme transmission during crossflow filtration of yeast suspensions using gas/liquid two-phase flows. AB - The optimal conditions for recovery of an enzyme were determined using gas/liquid two-phase flows. When filtering the enzyme-only solution under single-phase flow conditions, severe fouling occurred. This fouling was manifest as a decline in flux to less than 2% of the initial water flux and a decline in protein concentration in the permeate to 30% of its initial value, during a five-hour filtration period. When yeast cells were added under the same experimental conditions, enzyme transmission was maintained at 100% for the five-hour period and the enzyme mass flux was twofold higher. During gas-sparged microfiltration of the enzyme/yeast mixture in a permeate-recycling mode at the same liquid flow rate, gas/liquid slug flow strongly decreased the transmission of the enzyme (70% decrease), even though the permeate flux was improved (140% improvement). As a result, the mass flux of the enzyme was significantly reduced. However, with a bubble flow pattern, the permeate flux was 1.5 times higher and the transmission was maintained at a high level. The enzyme mass flux was then 25% higher when compared to single-phase flow filtration conditions. During diafiltration experiments with a bubble flow pattern, a 13% higher enzyme recovery was achieved. PMID- 12783840 TI - Turnup turndown of membrane operation of membrane bioreactors. AB - Membrane bioreactors can be operated with intermittent permeation and continuous aeration. Aeration close to the surface of a submerged membrane helps to maintain a membrane surface that is free from fouling. The conditions under which this occurs depend on the interaction between flux and aeration rate. Increased flux is possible without severe fouling if the aeration rate is increased. Results of performing membrane operation under the dual intermittency of aeration rate and permeation rate, with permeation also interrupted on a regular cycle, are presented. The results show that membrane plants designed for optimal operation at moderate flux can survive effectively with higher flux operation for restricted periods. Designing to account for such effects could reduce overall plant costs. PMID- 12783841 TI - Designing blood oxygenators. AB - Extracorporeal blood oxygenators are used to provide cardiopulmonary support during open heart surgery. In the study reported here, mass transfer correlations were determined for commercially available blood oxygenators. Two configurations used commercially, flow outside and across bundles of hollow fibers and flow in thin channels between parallel flat sheet membranes, were investigated. Water and glycerol/water mixtures were used as a substitute for blood. Diffusion of oxygen into and out of these solutions was studied. For flow across bundles of hollow fibers, the mass transfer correlations derived here are in agreement with analogous correlations for crossflow heat exchangers. However, for flow in thin channels, the rate of mass transfer is often less than predicted from theory. This compromised mass transfer can be explained by considering slight variations in the thickness of the blood flow channels. The mass transfer correlations developed here could be used to design better blood oxygenators. PMID- 12783842 TI - Membrane evaporative cooling to 30 degrees C or less: 1. Membrane evaporative cooling of contained water. AB - Microporous hydrophobic membranes have been examined for possible use as containers in the evaporative cooling of water, particularly in desert climates. An experimental determination was made of the overall heat and mass transfer coefficients of these membranes while surmounting contained water and with air flowing over the surface of the membranes. Similar tests were made with water alone, that is, without a membrane. The coefficients were then used to compare the performance of existing (canvas water) coolers and membrane evaporative coolers under desert conditions. The performance of the membrane coolers was close enough to that of the canvas coolers that extensive investigation of various aspects of membrane evaporative cooling appears to be justified, particularly in view of the potential advantages of the latter over the existing evaporative cooling methods. For example, for cool storage of perishable goods in a desert climate, the membrane container might be uniquely qualified because of its low rate of water consumption compared to that of a canvas cooler. PMID- 12783843 TI - Membrane evaporative cooling to 30 degrees C or less: 2. Membrane evaporative air cooling. AB - Microporous hydrophobic membranes were examined for use in steady state membrane evaporative air cooling. The examination consisted of calculating membrane performance as a function of overall heat and mass transfer coefficients already obtained and reported in Part 1 (previous paper, this volume). This performance was compared with that obtained by similar calculations made on existing evaporative air coolers. It was found that the cooling performance of the membrane evaporative air cooler was not as good as that of the existing evaporative air cooler. This is to be expected since the existing cooler has only one resistance, the air boundary layer (ABL), whereas the membrane cooler has the ABL and the membrane resistance. However, the membrane air cooler has advantages, such as appreciably lower water consumption and operation under more sanitary conditions, that is, without intimate conjunction of flowing air and liquid water on solid surfaces. PMID- 12783844 TI - Typical type 2 diabetes mellitus and HFE gene mutations: a population-based case control study. AB - Diabetes mellitus is a recognized consequence of hereditary haemochromatosis. Whether the common HFE mutations, that associate with this condition and pre dispose to increases in serum iron indices, are over-represented in diabetic populations remains controversial. We present data from the largest case-control study of the C282Y and H63D HFE allele frequencies in typical type 2 diabetes mellitus, as defined by an age of onset greater than 30 years and no requirement for insulin in the first year post-diagnosis. We also present a meta-analysis of all similar studies to date. We see no evidence for over-representation of iron loading HFE alleles in type 2 diabetes mellitus, suggesting that screening for HFE mutations in this population is of no value. PMID- 12783845 TI - Neuromuscular defects in a Drosophila survival motor neuron gene mutant. AB - Autosomal recessive spinal muscular atrophy (SMA) is linked to mutations in the survival motor neuron (SMN) gene. The SMN protein has been implicated at several levels of mRNA biogenesis and is expressed ubiquitously. Studies in various model organisms have shown that the loss of function of the SMN gene leads to embryonic lethality. The human contains two genes encoding for SMN protein and in patients one of these is disrupted. It is thought the remaining low levels of protein produced by the second SMN gene do not suffice and result in the observed specific loss of lower motor neurons and muscle wasting. The early lethality in the animal mutants has made it difficult to understand why primarily these tissues are affected. We have isolated a Drosophila smn mutant. The fly alleles contain point mutations in smn similar to those found in SMA patients. We find that zygotic smn mutant animals show abnormal motor behavior and that smn gene activity is required in both neurons and muscle to alleviate this phenotype. Physiological experiments on the fly smn mutants show that excitatory post synaptic currents are reduced while synaptic motor neuron boutons are disorganized, indicating defects at the neuromuscular junction. Clustering of a neurotransmitter receptor subunit in the muscle at the neuromuscular junction is severely reduced. This new Drosophila model for SMA thus proposes a functional role for SMN at the neuromuscular junction in the generation of neuromuscular defects. PMID- 12783846 TI - Polyglutamine protein aggregation and toxicity are linked to the cellular stress response. AB - Chronic exposure of cells to expanded polyglutamine proteins results in eventual cell demise. We constructed mouse cell lines expressing either the full-length androgen receptor (AR), or truncated forms of AR containing 25 or 65 glutamines to study the cellular consequences of chronic low-level exposure to these proteins. Expression of the polyglutamine-expanded truncated AR protein, but not the full-length expanded protein, resulted in the formation of cytoplasmic and nuclear aggregates and eventual cell death. Nuclear aggregates preferentially stained positive for heat shock protein (hsp)72, a sensitive indicator of a cellular stress response. Biochemical studies revealed that the presence of nuclear aggregates correlated with activation of the c-jun NH2-terminal kinase (JNK). Different metabolic insults, including heat shock treatment, and exposure to sodium arsenite or menadione, proved more toxic to those cells expressing the polyglutamine-expanded truncated protein than to cells expressing the non expanded form. Cells containing cytoplasmic polyglutamine-protein aggregates exhibited a delayed expression of hsp72 after heat shock. Once expressed, hsp72 failed to localize normally and instead was sequestered within the protein aggregates. This was accompanied by an inability of the aggregate-containing cells to cease their stress response as evidenced by the continued presence of activated JNK. Finally, activation of the cellular stress response increased the overall extent of polyglutamine protein aggregation, especially within the nucleus. Inclusion of a JNK inhibitor reduced this stress-dependent increase in nuclear aggregates. Abnormal stress responses may contribute to enhanced cell vulnerability in cells expressing polyglutamine-expanded proteins and may increase the propensity of such cells to form cytoplasmic and nuclear inclusions. PMID- 12783847 TI - Huntingtin contains a highly conserved nuclear export signal. AB - Huntington's disease (HD), is a genetic neurodegenerative disease characterized by a DNA CAG triplet repeat expansion in the first exon of the disease gene, HD. CAG DNA expansion results in a polyglutamine tract expansion in mutant huntingtin protein. Wild-type and mutant full-length huntingtin have been detected in the nucleus, but elevated levels of mutant huntingtin and huntingtin amino-terminal proteolytic fragments are seen to accumulate in the nuclei of HD-affected neurons. The presence of huntingtin in both the nucleus and the cytoplasm suggested that huntingtin may be dynamic between these compartments. By live cell time-lapse video microscopy, we have been able to visualize polyglutamine mediated aggregation and the transient nuclear localization of huntingtin over time in a striatal cell line. A classical nuclear localization signal could not be detected in huntingtin, but we have discovered a nuclear export signal (NES) in the carboxy-terminus of huntingtin. Leptomycin B treatment of clonal striatal cells enhanced the nuclear localization of huntingtin, and a mutant NES huntingtin displayed increased nuclear localization, indicating that huntingtin can shuttle to and from the nucleus. The huntingtin NES is strictly conserved among all huntingtin proteins from diverse species. This export signal may be important in Huntington's disease because this fragment of huntingtin is proteolytically cleaved away during HD. The huntingtin NES therefore defines a potential role for huntingtin as a member of a nucleocytoplasmic dynamic protein complex. PMID- 12783848 TI - Maternal alleles acquiring paternal methylation patterns in biparental complete hydatidiform moles. AB - We previously mapped a maternal locus responsible for biparental complete hydatidiform moles (BiCHMs) to 19q13.4. The two index patients had a total of 14 molar pregnancies, eight abortions at various developmental stages, and one 16 year-old healthy offspring. We suggested that the defective gene deregulates the expression of imprinted genes. Here, we report the methylation status of four imprinted genes in two BiCHMs from the two sisters, the 16-year-old normal offspring, and two sporadic BiCHMs from unrelated patients. Using two bisulfite based methods, we demonstrate a general trend of abnormal hypomethylation at the paternally expressed genes, PEG3 and SNRPN, and hypermethylation at the maternally expressed genes, NESP55 and H19, in two to four BiCHMs. Using single nucleotide polymorphisms, we provide the first evidence that SNRPN, NESP55 and H19 are abnormally methylated on the maternal alleles in BiCHMs. We show, in the BiCHMs from the two sisters, that the abnormally methylated H19 allele is inherited from either the maternal grandmother or the maternal grandfather. These data suggest that the abnormal methylation in BiCHMs is not due to an error in erasing the parental imprinting marks but rather in the re-establishment of the new maternal marks during oogenesis or their postzygotic maintenance. The defective 19q13.4 locus may have led to the development of variable degrees of 'faulty' paternal marks on the maternal chromosomes. PMID- 12783849 TI - IL1 receptor accessory protein like, a protein involved in X-linked mental retardation, interacts with Neuronal Calcium Sensor-1 and regulates exocytosis. AB - Previously, human genetics-based approaches allowed us to show that mutations in the IL-1 receptor accessory protein-like gene (IL1RAPL) are responsible for a non specific form of X-linked mental retardation. This gene encodes a predicted protein of 696 amino acids that belongs to a novel class of the IL-1/Toll receptor family. In addition to the extracellular portion consisting of three Ig like domains and the intracellular TIR domain characteristic of the IL-1/Toll receptor family, IL1RAPL contains a specific 150 amino acid carboxy terminus that has no significant homology with any protein of known function. In order to begin to elucidate the function of this IL-1/Toll receptor-like protein, we have assessed the effect of recombinant IL1RAPL on the binding affinity of type I IL 1R for its ligands IL-1alpha and beta and searched for proteins interacting with the specific carboxy terminus domain of IL1RAPL. Our results show that IL1RAPL is not a protein receptor for IL-1. In addition we present here the identification of Neuronal Calcium Sensor-1 (NCS-1) as an IL1RAPL interactor. Remarkably, although NCS-1 and its non-mammalian homologue, frequenin, are members of a highly conserved EF-hand Ca(2+) binding protein family, our data show that IL1RAPL interacts only with NCS-1 through its specific C-terminal domain. The functional relevance of IL1RAPL activity was further supported by the inhibitory effect on exocytosis in PC12 cells overexpressing IL1RAPL. Taken together, our data suggest that IL1RAPL may regulate calcium-dependent exocytosis and provide insight into the understanding of physiopathological mechanisms underlying cognitive impairment resulting from IL1RAPL dysfunction. PMID- 12783850 TI - The p38 subunit of the aminoacyl-tRNA synthetase complex is a Parkin substrate: linking protein biosynthesis and neurodegeneration. AB - Parkinson's disease (PD) is a severe neurological disorder, characterized by the progressive degeneration of the dopaminergic nigrostriatal pathway and the presence of Lewy bodies (LBs). The discovery of genes responsible for familial forms of the disease has provided insights into its pathogenesis. Mutations in the parkin gene, which encodes an E3 ubiquitin-protein ligase involved in the ubiquitylation and proteasomal degradation of specific protein substrates, have been found in nearly 50% of patients with autosomal-recessive early-onset parkinsonism. The abnormal accumulation of substrates due to loss of Parkin function may be the cause of neurodegeneration in parkin-related parkinsonism. Here, we demonstrate that Parkin interacts with, ubiquitylates and promotes the degradation of p38, a key structural component of the mammalian aminoacyl-tRNA synthetase complex. We found that the ubiquitylation of p38 is abrogated by truncated variants of Parkin lacking essential functional domains, but not by the pathogenic Lys161Asn point mutant. Expression of p38 in COS7 cells resulted in the formation of aggresome-like inclusions in which Parkin was systematically sequestered. In the human dopaminergic neuroblastoma-derived SH-SY5Y cell line, Parkin promoted the formation of ubiquitylated p38-positive inclusions. Moreover, the overexpression of p38 in SH-SY5Y cells caused significant cell death against which Parkin provided protection. Analysis of p38 expression in the human adult midbrain revealed strong immunoreactivity in normal dopaminergic neurons and the labeling of LBs in idiopathic PD. This suggests that p38 plays a role in the pathogenesis of PD, opening the way for a detailed examination of its potential non-canonical role in neurodegeneration. PMID- 12783851 TI - Loss of DNA-dependent dimerization of the transcription factor SOX9 as a cause for campomelic dysplasia. AB - Campomelic dysplasia (CD) is a semilethal osteochondrodysplasia, characterized by skeletal anomalies that include bending of the long bones, and by XY sex reversal. CD results from haploinsufficiency for the transcription factor SOX9, a key regulator at various steps of cartilage differentiation and of early testis development. Two functional domains are so far recognized for SOX9, a high mobility group (HMG) DNA-binding domain and a C-terminal transactivation domain. We present two CD patients with de novo mutations in a conserved region preceding the HMG domain. A long-term survivor with the acampomelic form of CD has an A76E amino acid substitution, while a severely affected CD patient had an in-frame deletion of amino acid residues 66-75. The conserved domain has been shown to function in the related transcription factor SOX10 as a DNA-dependent dimerization domain. We show that, like SOX10, SOX9 also binds cooperatively as a dimer to response elements in regulatory regions of some target genes such as the cartilage genes Col11a2 and CD-Rap. Dimerization and the resulting capacity to activate promoters via dimeric binding sites is lost in both mutant SOX9 proteins while other features involved in SOX9 function remained unaltered. These findings establish the dimerization domain as the third domain essential for SOX9 function during chondrogenesis. PMID- 12783852 TI - Fukutin is required for maintenance of muscle integrity, cortical histiogenesis and normal eye development. AB - Fukuyama-type congenital muscular dystrophy (FCMD), one of the most common autosomal-recessive disorders in Japan, is characterized by congenital muscular dystrophy associated with brain malformation due to a defect during neuronal migration. Through positional cloning, we previously identified the gene for FCMD, which encodes the fukutin protein. Here we report that chimeric mice generated using embryonic stem cells targeted for both fukutin alleles develop severe muscular dystrophy, with the selective deficiency of alpha-dystroglycan and its laminin-binding activity. In addition, these mice showed laminar disorganization of the cortical structures in the brain with impaired laminin assembly, focal interhemispheric fusion, and hippocampal and cerebellar dysgenesis. Further, chimeric mice showed anomaly of the lens, loss of laminar structure in the retina, and retinal detachment. These results indicate that fukutin is necessary for the maintenance of muscle integrity, cortical histiogenesis, and normal ocular development and suggest the functional linkage between fukutin and alpha-dystroglycan. PMID- 12783853 TI - Transcription-dependent recombination and the role of fork collision in yeast rDNA. AB - It is speculated that the function of the replication fork barrier (RFB) site is to avoid collision between the 35S rDNA transcription machinery and the DNA replication fork, because the RFB site is located near the 3'-end of the gene and inhibits progression of the replication fork moving in the opposite direction to the transcription machinery. However, the collision has never been observed in a blockless (fob1) mutant with 150 copies of rDNA. The gene FOB1 was shown previously to be required for replication fork blocking activity at the RFB site, and also for the rDNA copy number variation through unequal sister-chromatid recombination. This study documents the detection of fork collision in an fob1 derivative with reduced rDNA copy number (approximately 20) using two-dimensional agarose gel electrophoresis. This suggests that most of these reduced copies are actively transcribed. The collision was dependent on the transcription by RNA polymerase I. In addition, the transcription stimulated rDNA copy number variation, and the production of the extrachromosomal rDNA circles (ERCs), whose accumulation is thought to be a cause of aging. These results suggest that such a transcription-dependent fork collision induces recombination, and may function as a general recombination trigger for multiplication of highly transcribed single copy genes. PMID- 12783854 TI - Periodic repression by the bHLH factor Hes7 is an essential mechanism for the somite segmentation clock. AB - Hes7, a bHLH gene essential for somitogenesis, displays cyclic expression of mRNA in the presomitic mesoderm (PSM). Here, we show that Hes7 protein is also expressed in a dynamic manner, which depends on proteasome-mediated degradation. Spatial comparison revealed that Hes7 and Lunatic fringe (Lfng) transcription occurs in the Hes7 protein-negative domains. Furthermore, Hes7 and Lfng transcription is constitutively up-regulated in the absence of Hes7 protein and down-regulated by stabilization of Hes7 protein. Thus, periodic repression by Hes7 protein is critical for the cyclic transcription of Hes7 and Lfng, and this negative feedback represents a molecular basis for the segmentation clock. PMID- 12783855 TI - Elimination of Mcl-1 is required for the initiation of apoptosis following ultraviolet irradiation. AB - Ultraviolet (UV) irradiation of HeLa cells triggers an apoptotic response mediated by mitochondria. Biochemical analysis of this response revealed that the elimination of cytosolic inhibitors is required for mitochondrial release of cytochrome c and subsequent caspase activation. These inhibitors were found to be Mcl-1 and Bcl-xL, two antiapoptotic members of the Bcl-2 family. Following UV treatment, Mcl-1 protein synthesis is blocked, the existing pool of Mcl-1 protein is rapidly degraded by the proteasome, and cytosolic Bcl-xL translocates to the mitochondria. These events are sequential; the elimination of Mcl-1 is required for the translocation of Bcl-xL. The disappearance of Mcl-1 is also required for other mitochondrial apoptotic events including Bax translocation, cytochrome c release, and caspase activation. PMID- 12783856 TI - Control of landmark events in meiosis by the CDK Cdc28 and the meiosis-specific kinase Ime2. AB - Meiosis is thought to require the protein kinase Ime2 early for DNA replication and the cyclin-dependent kinase Cdc28 late for chromosome segregation. To elucidate the roles of these kinases, we inhibited their activities early and late using conditional mutants that are sensitive to chemical inhibitors. Our studies reveal that both Cdc28 and Ime2 have critical roles in meiotic S phase and M phase. Early inhibition of analog-sensitive cdc28-as1 blocked DNA replication, revealing a previously undetected role for Cdc28. Yet Cdc28 was dispensable for one of its functions in the mitotic cell cycle, degradation of Sic1. Late addition of inhibitor to ime2-as1 revealed unexpected roles of Ime2 in the initiation and execution of chromosome segregation. The requirement of Ime2 for M phase is partially explained by its stimulation of the key meiotic transcription factor Ndt80, which is needed in turn for high Cdc28 activity. In accordance with a late role for Ime2, we observed an increase in its activity during M phase that depended on Cdc28 and Ndt80. We speculate that several unique features of the meiotic cell division reflect a division of labor and regulatory coordination between Ime2 and Cdc28. PMID- 12783857 TI - Studies on the internalization mechanism of cationic cell-penetrating peptides. AB - A great deal of data has been amassed suggesting that cationic peptides are able to translocate into eucaryotic cells in a temperature-independent manner. Although such peptides are widely used to promote the intracellular delivery of bioactive molecules, the mechanism by which this cell-penetrating activity occurs still remains unclear. Here, we present an in vitro study of the cellular uptake of peptides, originally deriving from protegrin (the SynB peptide vectors), that have also been shown to enhance the transport of drugs across the blood-brain barrier. In parallel, we have examined the internalization process of two lipid interacting peptides, SynB5 and pAntp-(43-58), the latter corresponding to the translocating segment of the Antennapedia homeodomain. We report a quantitative study of the time- and dose-dependence of internalization and demonstrate that these peptides accumulate inside vesicular structures. Furthermore, we have examined the role of endocytotic pathways in this process using a variety of metabolic and endocytosis inhibitors. We show that the internalization of these peptides is a temperature- and energy-dependent process and that endosomal transport is a key component of the mechanism. Altogether, our results suggest that SynB and pAntp-(43-58) peptides penetrate into cells by an adsorptive mediated endocytosis process rather than temperature-independent translocation. PMID- 12783858 TI - Protein profiling with Epstein-Barr nuclear antigen-1 reveals an interaction with the herpesvirus-associated ubiquitin-specific protease HAUSP/USP7. AB - The Epstein-Barr nuclear antigen-1 (EBNA1) protein of Epstein-Barr virus is important for the replication, segregation, and transcriptional activation of latent Epstein-Barr virus genomes; has been implicated in host cell immortalization; and avoids proteasomal processing and cell-surface presentation. To gain insight into how EBNA1 fulfills these functions, we have profiled cellular protein interactions with EBNA1 using EBNA1 affinity chromatography and tandem affinity purification (TAP) of EBNA1 complexes from human cells (TAP tagging). We discovered several new specific cellular protein interactions with EBNA1, including interactions with HAUSP/USP7, NAP1, template-activating factor-I beta/SET, CK2, and PRMT5, all of which play important cell regulatory roles. The ubiquitin-specific protease USP7 is a known target of herpes simplex virus, and the USP7-binding region of EBNA1 was mapped to amino acids 395-450. A mutation in EBNA1 that selectively disrupted binding to USP7 was found to cause a 4-fold increase in EBNA1 replication activity but had no effect on EBNA1 turnover and cell-surface presentation. The results suggest that USP7 can regulate the replication function of EBNA1 and that EBNA1 may influence cellular events by sequestering key regulatory proteins. PMID- 12783859 TI - ATP binding to the first nucleotide binding domain of multidrug resistance associated protein plays a regulatory role at low nucleotide concentration, whereas ATP hydrolysis at the second plays a dominant role in ATP-dependent leukotriene C4 transport. AB - Multidrug resistance-associated protein (MRP1) transports solutes in an ATP dependent manner by utilizing its two nonequivalent nucleotide binding domains (NBDs) to bind and hydrolyze ATP. The two NBDs possess different properties (Gao, M., Cui, H. R., Loe, D. W., Grant, C. E., Almquist, K. C., Cole, S. P., and Deeley, R. G. (2000) J. Biol. Chem. 275, 13098-13108; Hou, Y., Cui, L., Riordan, J. R., and Chang, X. (2000) J. Biol. Chem. 275, 20280-20287) and may play different roles during solute transport. We now report that NBD1 has moderately higher affinity for ATP than NBD2. The consequence of this difference is that the overall Kd value for wild-type MRP1 is mainly determined by ATP binding at NBD1. This conclusion is supported by the following: 1) mutation of the cysteine residue at 682 to alanine (C682A) in Walker A motif in NBD1 decreases the Kd value, indicating increased affinity for ATP; 2) mutation of the alanine residue at 1331 to cysteine (A1331C) in the Walker A motif of NBD2 does not have an effect on the Kd value; and 3) photolabeling of the protein with a cysteine residue in the Walker A motif of NBD1 is much more sensitive to N-ethylmaleimide modification than the protein with a cysteine residue in the Walker A motif of NBD2. In contrast, the Km for ATP in support of LTC4 transport is mainly determined by ATP hydrolysis at NBD2. This conclusion is supported by the following: 1) although mutation of A1331C does not have an effect on the Kd value, the Km values measured from LTC4 transport by proteins with this mutation in NBD2 are much higher than the proteins with wild-type NBD2, implying that the A1331C mutation affects ATP binding/hydrolysis at NBD2; and 2) ATP-dependent LTC4 transport by the protein with a cysteine residue in the Walker A motif of NBD2 is much more sensitive to N-ethylmaleimide modification than the protein with a cysteine residue in the Walker A motif of NBD1. Our previous results indicated that ATP binding at NBD1 at low concentration enhanced ATP binding/hydrolysis at NBD2. All of these results support the notion that ATP binding at NBD1 at low concentration plays a more important regulatory role than the binding at high ATP concentration and that ATP hydrolysis at NBD2 plays a dominant role in the ATP dependent LTC4 transport. PMID- 12783861 TI - A maurotoxin with constrained standard disulfide bridging: innovative strategy of chemical synthesis, pharmacology, and docking on K+ channels. AB - Maurotoxin (MTX) is a 34-residue toxin that has been isolated initially from the venom of the scorpion Scorpio maurus palmatus. It presents a large number of pharmacological targets, including small conductance Ca2+-activated and voltage gated K+ channels. Contrary to other toxins of the alpha-KTx6 family (Pi1, Pi4, Pi7, and HsTx1), MTX exhibits a unique disulfide bridge organization of the type C1-C5, C2-C6, C3-C4, and C7-C8 (instead of the conventional C1-C5, C2-C6, C3-C7, and C4-C8, herein referred to as Pi1-like) that does not prevent its folding along the classic alpha/beta scaffold of scorpion toxins. Here, we developed an innovative strategy of chemical peptide synthesis to produce an MTX variant (MTXPi1) with a conventional pattern of disulfide bridging without any alteration of the toxin chemical structure. This strategy was used solely to address the impact of half-cystine pairings on MTX structural properties and pharmacology. The data indicate that MTXPi1 displays some marked changes in affinities toward the target K+ channels. Computed docking analyses using molecular models of both MTXPi1 and the various voltage-gated K+ channel subtypes (Shaker B, Kv1.2, and Kv1.3) were found to correlate with MTXPi1 pharmacology. A functional map detailing the interaction between MTXPi1 and Shaker B channel was generated in line with docking experiments. PMID- 12783860 TI - Understanding human cancer using Drosophila: Tid47, a cytosolic product of the DnaJ-like tumor suppressor gene l2Tid, is a novel molecular partner of patched related to skin cancer. AB - Recessive mutations of the Drosophila gene lethal(2)-tumorous imaginal discs (l(2)tid) cause neoplastic growth of the anlagen of the adult organs, the imaginal discs. Here we report that the three proteins encoded by this evolutionarily conserved gene, Tid50, Tid47, and Tid40, identified as members of the DnaJ cochaperone family, are destined for different cellular compartments, build complexes with many proteins in a developmental stage-specific manner, and are likely to be involved in different cellular processes. We show that the cytosolic Tid47 molecule is a novel component of the Hedgehog (Hh)-Patched (Ptc) signaling regulating cell/tissue polarity and spatial patterning during development and is associated with human tumors such as basal cell carcinoma (BCC) and medulloblastoma. We provide functional evidence for its direct in vivo interaction with the Hh-bound Ptc receptor during signal transmission. Because loss of l(2)tid causes neoplastic transformation of Hh-responsive cells, we suggest that Tid47 may at least act as a guardian of the Hh signaling gradient by regulating Ptc homeostasis in the tissue. Finally, we show that the expression of htid-1, the human counterpart of l(2)tid, is altered in human BCCs. We demonstrate that in BCCs loss of htid expression correlates with loss of differentiation capacity of the neoplastic cells similar to that found in the Drosophila tumor model. PMID- 12783862 TI - The SRC homology 2 domain of Rin1 mediates its binding to the epidermal growth factor receptor and regulates receptor endocytosis. AB - Activated epidermal growth factor receptors (EGFRs) recruit intracellular proteins that mediate receptor signaling and endocytic trafficking. Rin1, a multifunctional protein, has been shown to regulate EGFR internalization (1). Here we show that EGF stimulation induces a specific, rapid, and transient membrane recruitment of Rin1 and that recruitment is dependent on the Src homology 2 (SH2) domain of Rin1. Immunoprecipitation of EGFR is accompanied by co immunoprecipitation of Rin1 in a time- and ligand-dependent manner. Association of Rin1 and specifically the SH2 domain of Rin1 with the EGFR was dependent on tyrosine phosphorylation of the intracellular domain of the EGFR. The recruitment of Rin1, observed by light microscopy, indicated that although initially cytosolic, Rin1 was recruited to both plasma membrane and endosomes following EGF addition. Moreover, the expression of the SH2 domain of Rin1 substantially impaired the internalization of EGF without affecting internalization of transferrin. Finally, we found that Rin1 co-immunoprecipitated with a number of tyrosine kinase receptors but not with cargo endocytic receptors. These results indicate that Rin1 provides a link via its SH2 domain between activated tyrosine kinase receptors and the endocytic pathway through the recruitment and activation of Rab5a. PMID- 12783863 TI - Novel antioxidant role of alcohol dehydrogenase E from Escherichia coli. AB - Alcohol dehydrogenase E (AdhE) is an Fe-enzyme that, under anaerobic conditions, is involved in dissimilation of glucose. The enzyme is also present under aerobic conditions, its amount is about one-third and its activity is only one-tenth of the values observed under anaerobic conditions. Nevertheless, its function in the presence of oxygen remained ignored. The data presented in this paper led us to propose that the enzyme has a protective role against oxidative stress. Our results indicated that cells deleted in adhE gene could not grow aerobically in minimal media, were extremely sensitive to oxidative stress and showed division defects. In addition, compared with wild type, mutant cells displayed increased levels of internal peroxides (even higher than those found in a Delta katG strain) and increased protein carbonyl content. This pleiotropic phenotype disappeared when the adhE gene was reintroduced into the defective strain. The purified enzyme was highly reactive with hydrogen peroxide (with a Ki of 5 microM), causing inactivation due to a metal-catalyzed oxidation reaction. It is possible to prevent this reactivity to hydrogen peroxide by zinc, which can replace the iron atom at the catalytic site of AdhE. This can also be achieved by addition of ZnSO4 to cell cultures. In such conditions, addition of hydrogen peroxide resulted in reduced cell viability compared with that obtained without the Zn treatment. We therefore propose that AdhE acts as a H2O2 scavenger in Escherichia coli cells grown under aerobic conditions. PMID- 12783864 TI - Regulation of cell proliferation by autocrine motility factor/phosphoglucose isomerase signaling. AB - Autocrine motility factor (AMF)/phosphoglucose isomerase (PGI; EC 5.3.1.9) is a housekeeping cytosolic enzyme that plays a key role in both glycolysis and gluconeogenesis pathways. AMF/PGI is also a multifunctional protein that displays cytokine properties, eliciting mitogenic, motogenic, and differentiation activities, and has been implicated in tumor progression and metastasis. Because little is known about AMF/PGI-dependent signaling in general and during tumorigenesis in particular, we sought to study its effect on the cell cycle. To elucidate the functional role of PGI, we stably transfected its cDNA into NIH/3T3 and BALB/c 3T3-A31 fibroblasts. Ectopic overexpression of PGI results in the acquisition of a transformed phenotype associated with an acceleration of G1 to S cell cycle transition. These were manifested by up-regulation of cyclin D1 expression and cyclin-dependent kinase activity and down-regulation of the cyclin dependent kinase inhibitor p27Kip1. The reduced p27Kip1 protein expression level in PGI-overexpressing cells could be restored to control levels by treatment with proteasome inhibitor. PGI-overexpressing cells also exhibited elevated expression of Skp2 involved in p27Kip1 ubiquitination and elevation in the levels of retinoblastoma protein hyperphosphorylation. Thus, we may conclude that the overexpression of AMF/PGI enhances cell proliferation together with up-regulation of cyclin/cyclin-dependent kinase activities and down-regulation of p27Kip1, whereas the induction of 3T3 fibroblast transformation by PGI is regulated by the retinoblastoma protein pathway. PMID- 12783865 TI - Identification of organelles in bacteria similar to acidocalcisomes of unicellular eukaryotes. AB - Acidocalcisomes are acidic calcium storage compartments described in several unicellular eukaryotes, including trypanosomatid and apicomplexan parasites, algae, and slime molds. In this work, we report that the volutin granules of Agrobacterium tumefaciens possess properties similar to the acidocalcisomes. Transmission electron microscopy revealed that each intracellular granule was surrounded by a membrane. X-ray microanalysis of the volutin granules showed large amounts of phosphorus, magnesium, potassium, and calcium. Calcium in the volutin granules increased when the bacteria were incubated at high extracellular calcium concentration. Immunofluorescence and immunoelectron microscopy, using antisera raised against peptide sequences conserved in the A. tumefaciens proton pyrophosphatase, indicated localization in intracellular vacuoles. Purification of the volutin granules using iodixanol density gradients indicated a preferential localization of the pyrophosphatase activity in addition to high concentrations of phosphate, pyrophosphate, short- and long-chain polyphosphate, but lack of markers of the plasma membrane. The pyrophosphatase activity was potassium-insensitive and inhibited by the pyrophosphate analogs, amynomethylenediphosphonate and imidodiphosphate, by dicyclohexylcarbodiimide, and by the thiol reagent N-ethylmaleimide. Polyphosphate was also localized to the volutin granules by 4',6'-diamino-2-phenylindole staining. The organelles were acidic, as demonstrated by staining with LysoSensor blue DND-167, a dye especially used to detect very acidic compartments in cells, and cycloprodigiosin, a compound isolated from a marine bacterium that has been shown to uncouple proton pyrophosphatase activity acting as a chloride/proton symport. The results suggest that acidocalcisomes arose before the prokaryotic and eukaryotic lineages diverged. PMID- 12783866 TI - The role of yeast DNA 3'-phosphatase Tpp1 and rad1/Rad10 endonuclease in processing spontaneous and induced base lesions. AB - Tpp1 is a DNA 3'-phosphatase in Saccharomyces cerevisiae that is believed to act during strand break repair. It is homologous to one domain of mammalian polynucleotide kinase/3'-phosphatase. Unlike in yeast, we found that Tpp1 could confer resistance to methylmethane sulfonate when expressed in bacteria that lack abasic endonuclease/3'-phosphodiesterase function. This species difference was due to the absence of delta-lyase activity in S. cerevisiae, since expression of bacterial Fpg conferred Tpp1-dependent resistance to methylmethane sulfonate in yeast lacking the abasic endonucleases Apn1 and Apn2. In contrast, beta-only lyases increased methylmethane sulfonate sensitivity independently of Tpp1, which was explained by the inability of Tpp1 to cleave 3' alpha,beta-unsaturated aldehydes. In parallel experiments, mutations of TPP1 and RAD1, encoding part of the Rad1/Rad10 3'-flap endonuclease, caused synthetic growth defects in yeast strains lacking Apn1. In contrast, Fpg expression led to a partial rescue of apn1 apn2 rad1 synthetic lethality by converting lesions into Tpp1-cleavable 3' phosphates. The collected experiments reveal a profound toxicity of strand breaks with irreparable 3' blocking lesions, and extend the function of the Rad1/Rad10 salvage pathway to 3'-phosphates. They further demonstrate a role for Tpp1 in repairing endogenously created 3'-phosphates. The source of these phosphates remains enigmatic, however, because apn1 tpp1 rad1 slow growth could be correlated with neither the presence of a yeast delta-lyase, the activity of the 3'-phosphate-generating enzyme Tdp1, nor levels of endogenous oxidation. PMID- 12783867 TI - Growth factor receptor-binding protein 10 (Grb10) as a partner of phosphatidylinositol 3-kinase in metabolic insulin action. AB - The regulation of the metabolic insulin response by mouse growth factor receptor binding protein 10 (Grb10) has been addressed in this report. We find mouse Grb10 to be a critical component of the insulin receptor (IR) signaling complex that provides a functional link between IR and p85 phosphatidylinositol (PI) 3-kinase and regulates PI 3-kinase activity. This regulatory mechanism parallels the established link between IR and p85 via insulin receptor substrate (IRS) proteins. A direct association was demonstrated between Grb10 and p85 but was not observed between Grb10 and IRS proteins. In addition, no effect of mouse Grb10 was observed on the association between IRS-1 and p85, on IRS-1-associated PI 3 kinase activity, or on insulin-mediated activation of IR or IRS proteins. A critical role of mouse Grb10 was observed in the regulation of PI 3-kinase activity and the resulting metabolic insulin response. Dominant-negative Grb10 domains, in particular the SH2 domain, eliminated the metabolic response to insulin in differentiated 3T3-L1 adipocytes. This was consistently observed for glycogen synthesis, glucose and amino acid transport, and lipogenesis. In parallel, the same metabolic responses were substantially elevated by increased levels of Grb10. A similar role of Grb10 was confirmed in mouse L6 cells. In addition to the SH2 domain, the Pro-rich amino-terminal region of Grb10 was implicated in the regulation of PI 3-kinase catalytic activity. These regulatory roles of Grb10 were extended to specific insulin mediators downstream of PI 3 kinase including PKB/Akt, glycogen synthase kinase, and glycogen synthase. In contrast, a regulatory role of Grb10 in parallel insulin response pathways including p70 S6 kinase, ubiquitin ligase Cbl, or mitogen-activated protein kinase p38 was not observed. The dissection of the interaction of mouse Grb10 with p85 and the resulting regulation of PI 3-kinase activity should help elucidate the complexity of the IR signaling mechanism. PMID- 12783868 TI - Nat3p and Mdm20p are required for function of yeast NatB Nalpha-terminal acetyltransferase and of actin and tropomyosin. AB - NatB Nalpha-terminal acetyltransferase of Saccharomyces cerevisiae acts cotranslationally on proteins with Met-Glu- or Met-Asp- termini and subclasses of proteins with Met-Asn- and Met-Met- termini. NatB is composed of the interacting Nat3p and Mdm20p subunits, both of which are required for acetyltransferase activity. The phenotypes of nat3-Delta and mdm20-Delta mutants are identical or nearly the same and include the following: diminished growth at elevated temperatures and on hyperosmotic and nonfermentable media; diminished mating; defective actin cables formation; abnormal mitochondrial and vacuolar inheritance; inhibition of growth by DNA-damaging agents such as methyl methanesulfonate, bleomycin, camptothecin, and hydroxyurea; and inhibition of growth by the antimitotic drugs benomyl and thiabendazole. The similarity of these phenotypes to the phenotypes of certain act1 and tpm1 mutants suggests that such multiple defects are caused by the lack of acetylation of actin and tropomyosins. However, the lack of acetylation of other unidentified proteins conceivably could cause the same phenotypes. Furthermore, unacetylated actin and certain N-terminally altered actins have comparable defective properties in vitro, particularly actin-activated ATPase activity and sliding velocity. PMID- 12783869 TI - Adhesion regulation of stromal cell-derived factor-1 activation of ERK in lymphocytes by phosphatases. AB - We have investigated whether chemokine signaling to the extracellular-signal regulated kinase (ERK) was regulated by beta 1-integrin-mediated adhesion in B- and T-cell lines. Activation of ERK by the chemokine SDF-1 can be regulated by adhesion to beta 1-integrin substrates in the T-cell lines MOLT-3, Jurkat, and H9 and in the Daudi B-cell line. In Jurkat T-cells, adhesion to the immobilized alpha 4 beta 1-integrin ligand VCAM-1 or to the alpha 5 beta 1-integrin ligand fibronectin regulated stromal-cell derived factor-1 (SDF-1) activation of ERK. Adhesion control of SDF-1 signaling was a rapid event, occurring as early as 10 min after adhesion, and loss of signaling occurred within 10 min of deadhesion. In contrast, SDF-1 activation of the ERK kinase MEK was independent of adhesion. Partial restoration of signaling to ERK in suspension was accomplished by pretreatment with pharmacological inhibitors of serine/threonine or protein tyrosine phosphatases. In addition, we used a non-radioactive phosphatase assay using phosphorylated ERK as the substrate to determine relative ERK dephosphorylation in whole cell extracts. These results showed greater relative ERK dephosphorylation in extracts from Jurkat cells treated in suspension, as compared with adherent cells. Therefore, these data suggest that adhesion influences SDF-1 activation of ERK by regulating the activity of ERK phosphatases. This identifies a novel locus of adhesion regulation of the ERK cascade. PMID- 12783870 TI - Recognition of arylsulfatase A and B by the UDP-N-acetylglucosamine:lysosomal enzyme N-acetylglucosamine-phosphotransferase. AB - The critical step for sorting of lysosomal enzymes is the recognition by a Golgi located phosphotransferase. The topogenic structure common to all lysosomal enzymes essential for this recognition is still not well defined, except that lysine residues seem to play a critical role. Here we have substituted surface located lysine residues of lysosomal arylsulfatases A and B. In lysosomal arylsulfatase A only substitution of lysine residue 457 caused a reduction of phosphorylation to 33% and increased secretion of the mutant enzyme. In contrast to critical lysines in various other lysosomal enzymes, lysine 457 is not located in an unstructured loop region but in a helix. It is not strictly conserved among six homologous lysosomal sulfatases. Based on three-dimensional structure comparison, lysines 497 and 507 in arylsulfatase B are in a similar position as lysine 457 of arylsulfatase A. Also, the position of oligosaccharide side chains phosphorylated in arylsulfatase A is similar in arylsulfatase B. Despite the high degree of structural homology between these two sulfatases substitution of lysines 497 and 507 in arylsulfatase B has no effect on the sorting and phosphorylation of this sulfatase. Thus, highly homologous lysosomal arylsulfatases A and B did not develop a single conserved phosphotransferase recognition signal, demonstrating the high variability of this signal even in evolutionary closely related enzymes. PMID- 12783871 TI - Cyclic AMP regulates aquaporin 5 expression at both transcriptional and post transcriptional levels through a protein kinase A pathway. AB - The membrane water channel aquaporin 5 (AQP5) plays an important role in transporting water across the apical surface of the alveolar epithelium and the epithelia of submucosal glands in the upper airway and nasopharynx. It is thus a potentially important target for modulating the fluid content of upper airway and nasopharyngeal secretions in disorders such as cystic fibrosis. Here, we have used an in vitro system to identify a molecular mechanism through which transcriptional and post-transcriptional regulation of AQP5 is accomplished. In a murine lung epithelial cell line (MLE-12), the addition of chlorophenylthio-cAMP (cpt-cAMP) causes a 4-fold increase in AQP5 mRNA and protein levels and induces translocation of AQP5 to the apical plasma membrane. Treatment with forskolin and isoproternol also caused similar increases in AQP5 expression both in vitro and in mouse lung tissue slices. The addition of actinomycin D abolished the cAMP mediated increase in AQP5 mRNA and showed that there was no increase in the half life of AQP5 mRNA, and inhibition of protein kinase A by H-89 blocked the cpt cAMP-mediated increase of AQP5. Pretreatment of cells with cycloheximide blocked the cpt-cAMP-mediated increase of AQP5 mRNA, indicating that de novo protein synthesis is essential for increased AQP5 transcription. Immunofluorescent micrographs of cells treated with cpt-cAMP showed a significantly stronger AQP5 signal at the plasma membrane as compared with untreated cells. These results show that cAMP regulates AQP5 at multiple levels, by increasing synthesis of AQP5 mRNA and by triggering translocation of AQP5 to the plasma membrane. PMID- 12783872 TI - Activation of JNK by Epac is independent of its activity as a Rap guanine nucleotide exchanger. AB - Guanine nucleotide exchange factors (GEFs) and their associated GTP-binding proteins (G-proteins) are key regulatory elements in the signal transduction machinery that relays information from the extracellular environment into specific intracellular responses. Among them, the MAPK cascades represent ubiquitous downstream effector pathways. We have previously described that, analogous to the Ras-dependent activation of the Erk-1/2 pathway, members of the Rho family of small G-proteins activate the JNK cascade when GTP is loaded by their corresponding GEFs. Searching for novel regulators of JNK activity we have identified Epac (exchange protein activated by cAMP) as a strong activator of JNK 1. Epac is a member of a growing family of GEFs that specifically display exchange activity on the Rap subfamily of Ras small G-proteins. We report here that while Epac activates the JNK severalfold, a constitutively active (G12V) mutant of Rap1b does not, suggesting that Rap-GTP is not sufficient to transduce Epac-dependent JNK activation. Moreover, Epac signaling to the JNKs was not blocked by inactivation of endogenous Rap, suggesting that Rap activation is not necessary for this response. Consistent with these observations, domain deletion mutant analysis shows that the catalytic GEF domain is dispensable for Epac mediated activation of JNK. These studies identified a region overlapping the Ras exchange motif domain as critical for JNK activation. Consistent with this, an isolated Ras exchange motif domain from Epac is sufficient to activate JNK. We conclude that Epac signals to the JNK cascade through a new mechanism that does not involve its canonical catalytic action, i.e. Rap-specific GDP/GTP exchange. This represents not only a novel way to activate the JNKs but also a yet undescribed mechanism of downstream signaling by Epac. PMID- 12783873 TI - JNK-interacting protein 1 promotes Akt1 activation. AB - Members of the JNK pathway are organized together by virtue of interactions with JNK interacting protein 1 (JIP1), a scaffold protein. Here we have investigated the possibility that JIP1 may also affect the catalytic activity of Akt1, a serine/threonine kinase that has been implicated in multiple cellular processes, including survival and proliferation. JIP1 expression enhanced Akt1 kinase activity in a dose-dependent manner following serum starvation in 293 cells. Cellular activation of Akt1 following stimulation with low concentrations of insulin-like growth factor (IGF-1) was elevated in the presence of JIP1. JIP1 expression also prolonged Akt1 stimulation after a short IGF-1 pulse. The mechanism of JIP1-mediated Akt1 activation involved JIP1 protein binding to the Akt1 pleckstrin homology domain, which in turn promoted the phosphorylation of the activation T-loop of Akt1 by phosphoinositide-dependent kinase-1. These results suggest that, in certain cellular contexts, JIP1 may act as an Akt1 scaffold, which regulates the enzymatic activity of Akt1. This study also indicates that JIP1 expression can exert signaling effects independent of JNK activity. PMID- 12783874 TI - The relationship between side reactions and slow inhibition of ribulose bisphosphate carboxylase revealed by a loop 6 mutant of the tobacco enzyme. AB - The first directed mutant of a higher plant ribulose-bisphosphate carboxylase/oxygenase (Rubisco), constructed by chloroplast transformation, is catalytically impaired but still able to support the plant's photosynthesis and growth (Whitney, S. M., von Caemmerer, S., Hudson, G. S., and Andrews, T. J. (1999) Plant Physiol. 121, 579-588). This mutant enzyme has a Leu to Val substitution at residue 335 in the flexible loop 6 of the large subunit, which closes over the substrate during catalysis. Its active site was intact, as judged by its barely impaired competency in the initial enolization step of the reaction sequence, and its ability to bind tightly the intermediate analog, 2'-carboxy-D arabinitol-1,5-bisphosphate. Prompted by observations that the mutant enzyme displayed much less slow inhibition during catalysis in vitro than the wild type, its tendency to catalyze side reactions and its response to the slow inhibitor D xylulose-1,5-bisphosphate were studied. The lessening in slow inhibition was not caused by reduced production of inhibitory side products. Except for pyruvate production, these reactions were strongly enhanced by the mutation, as was the ability to catalyze the carboxylation of D-xylulose-1,5-bisphosphate. Rather, reduced inhibition was the result of lessened sensitivity to these inhibitors. The slow isomerization phase that characterizes inhibition of the wild-type enzyme by D-xylulose-1,5-bisphosphate was completely eliminated by the mutation, and the mutant was more adept than the wild type in catalyzing the benzylic acid type rearrangement of D-glycero-2,3-pentodiulose-1,5-bisphosphate (produced by oxidation of the substrate, D-ribulose-1,5-bisphosphate). These observations are consistent with increased flexibility of loop 6 induced by the mutation, and they reveal the underlying mechanisms by which the side reactions cause slow inhibition. PMID- 12783875 TI - Cloning and characterization of a mouse endoplasmic reticulum alkaline ceramidase: an enzyme that preferentially regulates metabolism of very long chain ceramides. AB - Ceramidases deacylate ceramides, important intermediates in the metabolic pathway of sphingolipids. In this study, we report the cloning and characterization of a novel mouse alkaline ceramidase (maCER1) with a highly restricted substrate specificity. maCER1 consists of 287 amino acids, and it has a 28 and 32% identity to the Saccharomyces alkaline ceramidases (YPC1p and YDC1p) and the human alkaline phytoceramidase, respectively. Reverse transcriptase-PCR analysis demonstrated that maCER1 was predominantly expressed in skin. maCER1 was localized to the endoplasmic reticulum as revealed by immunocytochemistry. In vitro biochemical characterization determined that maCER1 hydrolyzed D-erythro ceramide exclusively but not D-erythro-dihydroceramide or D-ribo-phytoceramide. Similar to other alkaline ceramidases, maCER1 had an alkaline pH optimum of 8.0, and it was activated by Ca2+ but inhibited by Zn2+,Cu2+, and Mn2+. maCER1 was also inhibited by sphingosine, one of its products. Metabolic labeling studies showed that overexpression of maCER1 caused a decrease in the incorporation of radiolabeled dihydrosphingosine into ceramide and complex sphingolipids but led to a concomitant increase in sphingosine-1-P (S1P) in HeLa cells. Mass measurement showed that overexpression of maCER1 selectively lowered the cellular levels of D-erythro-C24:1-ceramide, but not other ceramide species and caused an increase in the levels of S1P. Taken together, these data suggest that maCER1 is a novel alkaline ceramidase with a stringent substrate specificity and that maCER1 is selectively expressed in skin and may have a role in regulating the levels of bioactive lipids ceramide and S1P, as well as complex sphingolipids. PMID- 12783876 TI - Crystal structure of the ectodomain of human FcalphaRI. AB - Human FcalphaRI (CD89) is the receptor specific for IgA, an immunoglobulin that is abundant in mucosa and is also found in high concentrations in serum. Although FcalphaRI is an immunoglobulin Fc receptor (FcR), it differs in many ways from FcRs for other immunoglobulin classes. The genes of most FcRs are located on chromosome 1 at 1q21-23, whereas FcalphaRI is on chromosome 19, at 19q13.4, a region called the leukocyte receptor complex, because it is clustered with several leukocyte receptor families including killer cell inhibitory receptors (KIRs) and leukocyte Ig-like receptors (LIRs). The amino acid sequence of FcalphaRI shares only 20% homology with other FcRs but it has around 35% homology with its neighboring LIRs and KIRs. In this work, we analyzed the crystal structure of the ectodomain of FcalphaRI and examined structure similarities between FcalphaRI and KIR2DL1, KIR2DL2 and LIR-1. Our data show that FcalphaRI, KIRs, and LIRs share a common hydrophobic core in their interdomain interface, and FcalphaRI is evolutionally closer to LIR than KIR. PMID- 12783877 TI - Interaction of T4 endonuclease V with DNA: importance of the flexible loop regions in protein-DNA interaction. AB - T4 endonuclease V (T4 endo V), a thymine dimer-specific DNA repair enzyme, and its interaction with DNA were investigated by nuclear magnetic resonance (NMR) spectroscopy. Backbone resonance assignment, chemical shift mapping, and 15N relaxation measurements were employed to the free and DNA-bound enzymes. The secondary structure and the tertiary fold of T4 endo V in solution were consistent with those from the crystallographic study. The backbone 1H and 15N chemical shift perturbation upon the addition of DNA without a lesion revealed that the residues including Arg3, Arg22-Arg26, Lys45-Phe60, and Lys86-Thr88 participate in DNA binding. However, when DNA with a lesion was added to the enzyme and concomitantly the catalytic reaction was completed, the resonances of Arg22, Glu23, and Arg26, which constitute the catalytic active site, and the resonance of Thr88, were perturbed in a different manner. The region around Lys45 Ser47 was found to be involved in DNA binding, which have not been reported elsewhere. The backbone relaxation measurements of the free and DNA-bound enzymes indicated that two loop regions, Lys45-Phe60 and Lys86-Asp92, show the high degree of backbone flexibility. These results imply that two flexible loop regions may play an important role in DNA binding and in scanning along DNA duplex to search the thymine dimer sites in UV-damaged DNA. PMID- 12783878 TI - Gene structure of the human metabotropic glutamate receptor 5 and functional analysis of its multiple promoters in neuroblastoma and astroglioma cells. AB - The metabotropic glutamate receptor 5 (mGluR5) has a discrete tissue expression mainly limited to neural cells. Expression of mGluR5 is developmentally regulated and undergoes dramatic changes in association with neuropathological disorders. We report the complete genomic structure of the mGluR5 gene, which is composed of 11 exons and encompasses approximately 563 kbp. Three clusters of multiple transcription initiation sites located on three distinct exons (IA, IB, and II), which undergo alternative splicing, have been identified. The 5'-flanking regions of these exons were isolated and, using a luciferase reporter gene assay, shown to possess active promoter elements in SKN-MC neuroblastoma and U178-MG astroglioma cells. Promoter IA was characterized by a CpG island; promoter IB contained a TATA box, and promoter II possessed three active Oct-1-binding sites. Preferential luciferase activity was observed in SKN-MC concomitant with differential DNA binding activity to several responsive elements, including CREB, Oct-1, C/EBP, and Brn-2. Exposure to growth factors produced enhanced expression of promoters IB and II in astroglioma cells and activation of NF-kappa B. These results suggest that alternative 5'-splicing and usage of multiple promoters may contribute regulatory mechanisms for tissue- and context-specific expression of the mGluR5 gene. PMID- 12783879 TI - Suppressor of cytokine signaling 3 regulates proliferation and activation of T helper cells. AB - Suppressors of cytokine signaling (SOCS) have been implicated in regulation of T cell activation and cytokine-mediated differentiation of T-helper cells. In this study we have characterized the pattern of SOCS expression in naive and activated primary T-helper cells, examined whether expression of SOCS genes is regulated by cytokine or T-cell receptor signaling, and analyzed the function of SOCS in differentiated T-cells. We show that SOCS1, SOCS2, SOCS3, CIS (cytokine-induced SH2 protein) genes are constitutively expressed in naive T-helper cells, with SOCS3 being the most abundant. Antigen stimulation of naive T-helper cells down regulates SOCS3 expression and concomitantly up-regulates SOCS1, SOCS2, and CIS gene transcription, suggesting that SOCS genes are regulated differentially by T cell activation. Down-regulation of SOCS3 expression is subsequently followed by gradual increase in SOCS3 level and corresponding decline in interleukin 2 (IL-2) secretion. In fact, SOCS3 mRNA levels are inversely correlated with the amount of IL-2 secretion and proliferative responses of differentiating T-helper cells, suggesting mutually antagonistic effects of SOCS3 and IL-2 and feedback regulation of T-cell activation by SOCS3. Furthermore, the degree of SOCS3 inhibition is antigen concentration-dependent and is mediated in part by growth factor independence-1, a T-cell transcription factor that regulates S-phase entry in T-cells. Forced overexpression of SOCS3 inhibits proliferation of T-helper cells, whereas depletion of endogenous SOCS3 by antisense SOCS3 cDNA enhances T cell receptor- and cytokine-induced proliferation. Taken together, these results suggest a role for SOCS3 in maintaining T-helper cells in a quiescent state. Transient inhibition of SOCS3 by antigen stimulation may therefore be essential in allowing activation of resting T-cells. PMID- 12783880 TI - Regulation of ERK1 and ERK2 by glucose and peptide hormones in pancreatic beta cells. AB - We showed previously that ERK1/2 were activated by glucose and amino acids in pancreatic beta cells. Here we examine and compare signaling events that are necessary for ERK1/2 activation by glucose and other stimuli in beta cells. We find that agents that interrupt Ca2+ signaling by a variety of mechanisms interfere with glucose- and glucagon-like peptide (GLP-1)-stimulated ERK1/2 activity. In particular, calmodulin antagonists, FK506, and cyclosporin, immunosuppressants that inhibit the calcium-dependent phosphatase calcineurin, suppress ERK1/2 activation by both glucose and GLP-1. Ca2+ signaling from intracellular stores is also essential for ERK1/2 activation, because thapsigargin blocks ERK1/2 activation by glucose or GLP-1. The glucose-sensitive mechanism is distinct from that used by phorbol ester or insulin to stimulate ERK1/2 but shares common features with that used by GLP-1. PMID- 12783881 TI - Identification of herpes simplex virus RNAs that interact specifically with regulatory protein ICP27 in vivo. AB - Herpes simplex virus type 1 (HSV-1) protein ICP27 has an essential regulatory role during viral replication, in part by post-transcriptional control of gene expression, and has a counterpart in all herpes viruses sequenced so far. Although much is known about the functions of this signature herpesvirus protein, little is known about its RNA binding capabilities; ICP27 interacts with specificity for a subset of intronless HSV-1 RNAs and poly(G), through its RGG box. We performed an in vivo yeast three-hybrid screen of an HSV-1 genomic library, searching for ICP27 interacting RNAs. Comparable with a yeast genomic screen, 24 of 55 single inserts mapped to antisense strands of HSV-1 transcribed regions or non-transcribed regions. The 31 HSV-1 sense RNAs identified were 35 to 225 nucleotides in length and interacted with preferred specificity for ICP27 as compared with an unrelated RNA-binding protein. They map to 10 monocistronic and 10 polycistronic transcripts of all kinetic classes and represent 28 open reading frames encoding predominantly essential viral proteins with roles in viral DNA replication and virion maturation. Several studies show regulatory effects by ICP27 on the majority of these transcripts, consistent with its regulation of the early-late switch in the HSV-1 life cycle. Deletion of the ICP27 RGG box and the ICP27 M15 mutation, both lethal in virus, abolished or severely reduced the ICP27 RNA interactions, indicating their biological relevance. The study facilitates continued study of gene regulation by ICP27 by further defining its interactions with viral RNAs. PMID- 12783882 TI - Rpn5 is a conserved proteasome subunit and required for proper proteasome localization and assembly. AB - Proper function of the 26 S proteasome requires assembly of the regulatory complex, which is composed of the lid and base subcomplexes. We characterized Rpn5, a lid subunit, in fission yeast. We show that Rpn5 associates with the proteasome rpn5. Deletion (rpn5Delta) exacerbates the growth defects in proteasome mutants, leading to mitotic abnormalities, which correlate with accumulation of polyubiquitinated proteins, such as Cut2/securin. Rpn5 expression is tightly controlled; both overexpression and deletion of rpn5 impair proteasome functions. The proteasome is assembled around the inner nuclear membrane in wild type cells; however, in rpn5Delta cells, proteasome subunits are improperly assembled and/or localized. In the lid mutants, Rpn5 is mislocalized in the cytosol, while in the base mutants, Rpn5 can enter the nucleus, but is left in the nucleoplasm, and not assembled into the nuclear membrane. These results suggest that Rpn5 is a dosage-dependent proteasome regulator and plays a role in mediating proper proteasome assembly. Moreover, the Rpn5 assembly may be a cooperative process that involves at least two steps: 1) nuclear import and 2) subsequent assembly into the nuclear membrane. The former step requires other components of the lid, while the latter requires the base. Human Rpn5 rescues the phenotypes associated with rpn5Delta and is incorporated into the yeast proteasome, suggesting that Rpn5 functions are highly conserved. PMID- 12783883 TI - K+-dependent cerebellar granule neuron apoptosis. Role of task leak K+ channels. AB - Rat mature cerebellar granule, unlike hippocampal neurons, die by apoptosis when cultured in a medium containing a physiological concentration of K+ but survive under high external K+ concentrations. Cell death in physiological K+ parallels the developmental expression of the TASK-1 and TASK-3 subunits that encode the pH sensitive standing outward K+ current IKso. Genetic transfer of the TASK subunits in hippocampal neurons, lacking IKso, induces cell death, while their genetic inactivation protects cerebellar granule neurons. Neuronal death of cultured rat granule neurons is also prevented by conditions that specifically reduce K+ efflux through the TASK-3 channels such as extracellular acidosis and ruthenium red. TASK leak K+ channels thus play an important role in K+-dependent apoptosis of cerebellar granule neurons in culture. PMID- 12783884 TI - Protein kinase B alpha/Akt1 regulates placental development and fetal growth. AB - Protein kinase B alpha (PKB alpha/Akt1) is implicated in the regulation of metabolism, transcription, cell survival, angiogenesis, cell migration, growth, and tumorigenesis. Previously, it was reported that PKB alpha-deficient mice are small with increased neonatal mortality (Cho, H., Thorvaldsen, J. L., Chu, Q., Feng, F., and Birnbaum, M. J. (2001) J. Biol. Chem. 276, 38349-38352 and Chen, W. S., Xu, P. Z., Gottlob, K., Chen, M. L., Sokol, K., Shiyanova, T., Roninson, I., Wenig, W., Suzuki, R., Tobe, K., Kadowaki, T., and Hay, N. (2001) Genes Dev. 15, 2203-2208). Here we show that PKB alpha is widely expressed in placenta including all types of trophoblast and vascular endothelial cells. Pkb alpha-/- placentae display significant hypotrophy, with marked reduction of the decidual basalis and nearly complete loss of glycogen-containing cells in the spongiotrophoblast, and exhibit decreased vascularization. Pkb alpha-/- placentae also show significant reduction of phosphorylation of PKB and endothelial nitric-oxide synthase. These defects may cause placental insufficiency, fetal growth impairment, and neonatal mortality. These data represent the first evidence for the role of PKB alpha and endothelial nitricoxide synthase in regulating placental development and provide an animal model for intrauterine growth retardation. PMID- 12783885 TI - Tyrosine phosphorylation disrupts elongin interaction and accelerates SOCS3 degradation. AB - The suppressors of cytokine signaling (SOCS) are negative feedback inhibitors of cytokine and growth factor-induced signal transduction. The C-terminal SOCS box region is thought to regulate SOCS protein stability most likely via an elongin C interaction. In the present study, we have found that phosphorylation of SOCS3 at two tyrosine residues in the conserved SOCS box, Tyr204 and Tyr221, can inhibit the SOCS3-elongin C interaction and activate proteasome-mediated SOCS3 degradation. Jak-mediated phosphorylation of SOCS3 decreased SOCS3 protein half life, and phosphorylation of both Tyr204 and Tyr221 was required to fully destabilize SOCS3. In contrast, a phosphorylation-deficient mutant of SOCS3, Y204F,Y221F, remained stable in the presence of activated Jak2 and receptor tyrosine kinases. SOCS3 stability correlated with the relative amount that bound elongin C, because in vitro phosphorylation of a SOCS3-glutathione S-transferase fusion protein abolished its ability to interact with elongin C. In addition, a SOCS3/SOCS1 chimera that co-precipitates with markedly increased elongin C, was significantly more stable than wild-type SOCS3. The data suggest that interaction with elongin C stabilizes SOCS3 protein expression and that phosphorylation of SOCS box tyrosine residues disrupts the complex and enhances proteasome-mediated degradation of SOCS3. PMID- 12783886 TI - Warm temperatures activate TRPV4 in mouse 308 keratinocytes. AB - Mammalian survival requires constant monitoring of environmental and body temperature. Recently, several members of the transient receptor potential vanilloid (TRPV) subfamily of ion channels have been identified that can be gated by increases in temperature into the warm (TRPV3 and TRPV4) or painfully hot (TRPV1 and TRPV2) range. In rodents, TRPV3 and TRPV4 proteins have not been detected in sensory neurons but are highly expressed in skin epidermal keratinocytes. Here, we show that in response to warm temperatures (>32 degrees C), the mouse 308 keratinocyte cell line exhibits nonselective transmembrane cationic currents and Ca2+ influx. Both TRPV3 and TRPV4 are expressed in 308 cells. However, the warmth-evoked responses we observe most closely resemble those mediated by recombinant TRPV4 on the basis of their electrophysiological properties and sensitivity to osmolarity and the phorbol ester, 4alpha-phorbol 12,13-didecanoate. Together, these data support the notion that keratinocytes are capable of detecting modest temperature elevations, strongly suggest that TRPV4 participates in these responses, and define a system for the cell biological analysis of warmth transduction. PMID- 12783887 TI - Nitric oxide formation by Escherichia coli. Dependence on nitrite reductase, the NO-sensing regulator Fnr, and flavohemoglobin Hmp. AB - Nitric oxide (NO) is a key signaling and defense molecule in biological systems. The bactericidal effects of NO produced, for example, by macrophages are resisted by various bacterial NO-detoxifying enzymes, the best understood being the flavohemoglobins exemplified by Escherichia coli Hmp. However, many bacteria, including E. coli, are reported to produce NO by processes that are independent of denitrification in which NO is an obligatory intermediate. We demonstrate using an NO-specific electrode that E. coli cells, grown anaerobically with nitrate as terminal electron acceptor, generate significant NO on adding nitrite. The periplasmic cytochrome c nitrite reductase (Nrf) is shown, by comparing Nrf+ and Nrf- mutants, to be largely responsible for NO generation. Surprisingly, an hmp mutant did not accumulate more NO but, rather, failed to produce detectable NO. Anaerobic growth of the hmp mutant was not stimulated by nitrate, and the mutant failed to produce periplasmic cytochrome(s) c, leading to the hypothesis that accumulating NO in the absence of Hmp inactivates the global anaerobic regulator Fnr by reaction with the [4Fe-4S]2+ cluster (Cruz-Ramos, H., Crack, J., Wu, G., Hughes, M. N., Scott, C., Thomson, A. J., Green, J., and Poole, R. K. (2002) EMBO J. 21, 3235-3244). Fnr thus failed to up-regulate nitrite reductase. The model is supported by the inability of an fnr mutant to generate NO and by the restoration of NO accumulation to hmp mutants upon introducing a plasmid encoding Fnr* (D154A) known to confer activity in the presence of oxygen. A cytochrome bd-deficient mutant retained NO-generating activity. The present study reveals a critical balance between NO-generating and -detoxifying activities during anaerobic growth. PMID- 12783888 TI - Regulation of c-myc gene by nitric oxide via inactivating NF-kappa B complex in P19 mouse embryonal carcinoma cells. AB - Nitric oxide (NO) may regulate gene expression by directly modifying redox state sensitive residues of transcription factors. Here we show that the NO donor, sodium nitroprusside (SNP), rapidly represses c-myc gene transcription in a protein synthesis-independent manner in P19 embryonal carcinoma cells by inactivation of NF-kappa B. SNP treatment reduces the DNA binding ability of the constitutively active NF-kappa B heterodimer, p65/p50, and its consequent transactivation of the c-myc promoter. Repression can be blocked by the peroxynitrite scavenger, deferoxamine, but not by dithiothreitol, which triggers reduction of S-nitrosylated residues. In HEK293 cells, where tumor necrosis factor-alpha can activate NF-kappa B, SNP likewise suppresses the binding of the active NF-kappa B complex, restoring the binding of the repressive p50/p50 homodimer complex. This effect of SNP in HEK293 cells is also blocked by deferoxamine. Chromatin immunoprecipitation analysis of SNP-treated P19 cells reveals reduced association of p65, but not of p50, with the promoter region of the endogenous c-myc gene. SNP-induced p65 dissociation was associated with the recruitment of histone deacetylase 1 and 2 to the endogenous c-myc gene promoter and the subsequent deacetylation of its chromatin histone. This study is the first to demonstrate that NO modulates the transcriptional activity of the c-myc gene promoter by dissociating the active form of NF-kappa B and replacing it with a repressive NF-kappa B complex, correlated with the recruitment of gene silencing histone deacetylases. In light of findings that NF-kappa B stimulates Myc oncoprotein expression in cancers, our findings suggest that NO should be investigated as a prospective therapeutic cancer agent. PMID- 12783890 TI - Hyperosmotic-induced protein kinase N 1 activation in a vesicular compartment is dependent upon Rac1 and 3-phosphoinositide-dependent kinase 1. AB - Protein kinase N 1 (PKN1), which in part resembles yeast protein kinase C, has been shown to be under the control of Rho GTPases and 3-phosphoinositide dependent kinase 1 (PDK1). We show here that green fluorescent protein-tagged PKN1 has the ability to translocate in a reversible manner to a vesicular compartment following hyperosmotic stress. PKN1 kinase activity is not necessary for this translocation, and in fact the PKN inhibitor HA1077 is also shown to induce PKN1 vesicle accumulation. PKN1 translocation is dependent on Rac1 activation, although the GTPase binding HR1abc domain is not sufficient for this recruitment. The PKN1 kinase domain, however, localizes constitutively to this compartment, and we demonstrate that this behavior is selective for PKNs. Associated with vesicle recruitment, PKN1 is shown to undergo activation loop phosphorylation and activation. It is established that this activation pathway involves PDK1, which is shown to be recruited to this PKN1-positive compartment upon hyperosmotic stress. Taken together, our findings present a pathway for the selective hyperosmotic-induced Rac1-dependent PKN1 translocation and PDK1 dependent activation. PMID- 12783889 TI - m-Calpain colocalizes with the calcium-sensing receptor (CaR) in caveolae in parathyroid cells and participates in degradation of the CaR. AB - The calcium-sensing receptor (CaR) is a G protein-coupled, seven-transmembrane receptor and resides within caveolin-rich membrane domains in bovine parathyroid cells. The proenzyme of calpain 2 (m-calpain) is a heterodimeric calcium dependent cysteine protease consisting of catalytic and regulatory subunits. The effects of calcium on the enzyme include activation, autolysis, and subunit dissociation. Here, we examine the potential role of caveolin-1 and caveolae in regulating the cellular distribution and function of m-calpain in parathyroid cells. We show that the inactive heterodimeric forms of m-calpain are concentrated in caveolin-rich membrane fractions prepared from parathyroid cells incubated with low extracellular calcium (Ca2+(o)). In contrast, in cells incubated with 3 mm Ca2+(o), which activates the CaR and increases intracellular calcium, there is a reduction in m-calpain in association with an increase in CaR protein and phosphorylated protein kinase C alpha and beta in caveolin-rich fractions. To assess the impact of activation of calpain on CaR protein in caveolar fractions, we analyzed the effects of m-calpain on the CaR. Activation of the CaR with high Ca2+(o) induced the release of lower molecular weight fragments of the receptor into the cell culture medium, and calpain inhibitors blocked this effect. Moreover, the fragments of the CaR as well as caveolin-1, m calpain, and alkaline phosphatase were localized in membrane vesicles shed by parathyroid cells, supporting the association of these proteins in living cells. Treatment of CaR proteins in vitro with m-calpain also resulted in the appearance of lower molecular weight fragments of the CaR. Our data suggest that localization of m-calpain within caveolae may contribute to maintenance of the enzyme in an inactive state and that m-calpain may also contribute to the regulation of CaR levels. PMID- 12783891 TI - Analysis of the open region and of DNA-protein contacts of archaeal RNA polymerase transcription complexes during transition from initiation to elongation. AB - The archaeal transcriptional machinery is polymerase II (pol II)-like but does not require ATP or TFIIH for open complex formation. We have used enzymatic and chemical probes to follow the movement of Pyrococcus RNA polymerase (RNAP) along the glutamate dehydrogenase gene during transcription initiation and transition to elongation. RNAP was stalled between registers +5 and +20 using C-minus cassettes. The upstream edge of RNAP was in close contact with the archaeal transcription factors TATA box-binding protein/transcription factor B in complexes stalled at position +5. Movement of the downstream edge of the RNAP was not detected by exonuclease III footprinting until register +8. A first structural transition characterized by movement of the upstream edge of RNAP was observed at registers +6/+7. A major transition was observed at registers +10/+11. In complexes stalled at these positions also the downstream edge of RNA polymerase started translocation, and reclosure of the initially open complex occurred indicating promoter clearance. Between registers +11 and +20 both RNAP and transcription bubble moved synchronously with RNA synthesis. The distance of the catalytic center to the front edge of the exo III footprint was approximately 12 nucleotides in all registers. The size of the RNA-DNA hybrid in an early archaeal elongation complex was estimated between 9 and 12 nucleotides. For complexes stalled between positions +10 and +20 the size of the transcription bubble was around 17 nucleotides. This study shows characteristic mechanistic properties of the archaeal system and also similarities to prokaryotic RNAP and pol II. PMID- 12783892 TI - hFis1, a novel component of the mammalian mitochondrial fission machinery. AB - The balance between the fission and fusion mechanisms regulate the morphology of mitochondria. In this study we have identified a mammalian protein that we call hFis1, which is the orthologue of the yeast Fis1p known to participate in yeast mitochondrial division. hFis1, when overexpressed in various cell types, localized to the outer mitochondrial membrane and induced mitochondrial fission. This event was inhibited by a dominant negative mutant of Drp1 (Drp1(K38A)), a major component of the fission apparatus. Fragmentation of the mitochondrial network by hFis1 was followed by the release of cytochrome c and ultimately apoptosis. Bcl-xL was able to block cytochrome c release and apoptosis but failed to prevent mitochondrial fragmentation. Our studies show that hFis1 is part of the mammalian fission machinery and suggest that regulation of the fission processes might be involved in apoptotic mechanisms. PMID- 12783894 TI - Differing roles of the N- and C-terminal zinc fingers in human immunodeficiency virus nucleocapsid protein-enhanced nucleic acid annealing. AB - The replication process of human immunodeficiency virus requires a number of nucleic acid annealing steps facilitated by the hybridization and helix destabilizing activities of human immunodeficiency virus nucleocapsid (NC) protein. NC contains two CCHC zinc finger motifs numbered 1 and 2 from the N terminus. The amino acids surrounding the CCHC residues differ between the two zinc fingers. Assays were preformed to investigate the activities of the fingers by determining the effect of mutant and wild-type proteins on annealing of 42 nucleotide RNA and DNA complements. The mutants 1.1 NC and 2.2 NC had duplications of the N- and C-terminal zinc fingers in positions 1 and 2. The mutant 2.1 NC had the native zinc fingers with their positions switched. Annealing assays were completed with unstructured and highly structured oligonucleotide complements. 2.2 NC had a near wild-type level of annealing of unstructured nucleic acids, whereas it was completely unable to stimulate annealing of highly structured nucleic acids. In contrast, 1.1 NC was able to stimulate annealing of both unstructured and structured substrates, but to a lesser degree than the wild-type protein. Results suggest that finger 1 has a greater role in unfolding of strong secondary structures, whereas finger 2 serves an accessory role that leads to a further increase in the rate of annealing. PMID- 12783895 TI - A host of novel agents for treating psoriasis, psoriatic arthritis stir interest. PMID- 12783893 TI - Inhibition of apoptosis by Z-VAD-fmk in SMN-depleted S2 cells. AB - Spinal muscular atrophy is an autosomal recessive motor neuron degenerative disorder, caused by the loss of telomeric copy of the survival motor neuron gene (SMN1). To better understand how motor neurons are targeted in Spinal muscular atrophy patients, it is important to study the role of SMN protein in cell death. In this report, we employed RNA interference (RNAi) to study the loss-of-function of SMN in Drosophila S2 cells. A 601-base pair double-stranded RNA (dsRNA) of Drosophila SMN (dSMN) was used for silencing the dSMN. Our data indicate that dSMN RNAi resulted in more than 90% reduction of both RNA and protein. Further analysis of S2 cells by cell death ELISA and flow cytometry assays revealed that reduction of dSMN expression significantly increased apoptosis. The cell death mediated by SMN depletion is caspase-dependent and specifically due to the activation of the endogenous caspases, DRONC and DRICE. Significantly, the effect of dSMN RNAi was reversed by a peptide caspase inhibitor, Z-VAD-fmk. These results suggest that dSMN is involved in signal pathways of apoptotic cell death in Drosophila. Hence, the model system of reduced SMN expression by RNAi in Drosophila could be exploited for identification of therapeutic targets. PMID- 12783896 TI - Researchers seek new uses for sildenafil. PMID- 12783897 TI - FDA issues warning on "all-natural" herbal product found to contain viagra. PMID- 12783898 TI - From the Centers for Disease Control and Prevention. Cluster of severe acute respiratory syndrome cases among protected health-care workers--Toronto, Canada, April 2003. PMID- 12783900 TI - From the Centers for Disease Control and Prevention. Update: severe acute respiratory syndrome--United States, May 14, 2003. PMID- 12783901 TI - Accurate measurement of blood pressure. PMID- 12783902 TI - Accurate measurement of blood pressure. PMID- 12783903 TI - Accurate measurement of blood pressure. PMID- 12783904 TI - Accurate measurement of blood pressure. PMID- 12783905 TI - Acetylcysteine and renal function following coronary angiographic procedures. PMID- 12783906 TI - Outcomes of medical vs invasive therapy for elderly patients with angina. PMID- 12783907 TI - Acetylcysteine and renal function following coronary angiographic procedures. PMID- 12783908 TI - Acetylcysteine and renal function following coronary angiographic procedures. PMID- 12783909 TI - Variation in use of broad-spectrum antibiotics for acute respiratory tract infection. PMID- 12783910 TI - Variation in use of broad-spectrum antibiotics for acute respiratory tract infection. PMID- 12783911 TI - Rapid magnetic resonance imaging vs radiographs for patients with low back pain: a randomized controlled trial. AB - CONTEXT: Faster magnetic resonance imaging (MRI) scanning has made MRI a potential cost-effective replacement for radiographs for patients with low back pain. However, whether rapid MRI scanning results in better patient outcomes than radiographic evaluation or a cost-effective alternative is unknown. OBJECTIVE: To determine the clinical and economic consequences of replacing spine radiographs with rapid MRI for primary care patients. DESIGN, SETTING, AND PATIENTS: Randomized controlled trial of 380 patients aged 18 years or older whose primary physicians had ordered that their low back pain be evaluated by radiographs. The patients were recruited between November 1998 and June 2000 from 1 of 4 imaging centers in the Seattle, Wash, area: a university-based teaching program, a nonuniversity-based teaching program, and 2 private clinics. INTERVENTION: Patients were randomly assigned to receive lumbar spine evaluation by rapid MRI or by radiograph. MAIN OUTCOME MEASURES: Back-related disability measured by the modified Roland questionnaire. Secondary outcomes included Medical Outcomes Study 36-Item Short Form Health Survey (SF-36), pain, preference scores, satisfaction, and costs. RESULTS: At 12 months, primary outcomes of functional disability were obtained from 337 (89%) of the 380 patients enrolled. The mean back-related disability modified Roland score for the 170 patients assigned to the radiograph evaluation group was 8.75 vs 9.34 for the 167 patients assigned the rapid MRI evaluation group (mean difference, -0.59; 95% CI, -1.69 to 0.87). The mean differences in the secondary outcomes were not statistically significant : pain bothersomeness (0.07; 95% CI -0.88 to 1.22), pain frequency (0.12; 95% CI, -0.69 to 1.37), and SF-36 subscales of bodily pain (1.25; 95% CI, -4.46 to 4.96), and physical functioning (2.73, 95% CI -4.09 to 6.22). Ten patients in the rapid MRI group vs 4 in the radiograph group had lumbar spine operations (risk difference, 0.34; 95% CI, -0.06 to 0.73). The rapid MRI strategy had a mean cost of 2380 dollars vs 2059 dollars for the radiograph strategy (mean difference, 321 dollars; 95% CI, -1100 to 458). CONCLUSIONS: Rapid MRIs and radiographs resulted in nearly identical outcomes for primary care patients with low back pain. Although physicians and patients preferred the rapid MRI, substituting rapid MRI for radiographic evaluations in the primary care setting may offer little additional benefit to patients, and it may increase the costs of care because of the increased number of spine operations that patients are likely to undergo. PMID- 12783912 TI - Effects of rofecoxib or naproxen vs placebo on Alzheimer disease progression: a randomized controlled trial. AB - CONTEXT: Laboratory evidence that inflammatory mechanisms contribute to neuronal injury in Alzheimer disease (AD), along with epidemiological evidence, suggests that nonsteroidal anti-inflammatory drugs (NSAIDs) may favorably influence the course of the disease. OBJECTIVE: To determine whether treatment with a selective cyclooxygenase (COX) -2 inhibitor (rofecoxib) or a traditional nonselective NSAID (naproxen) slows cognitive decline in patients with mild-to-moderate AD. DESIGN: Multicenter, randomized, double-blind, placebo-controlled, parallel group trial, with 1-year exposure to study medications. SETTING: Forty ambulatory treatment centers affiliated with the Alzheimer's Disease Cooperative Study consortium. PARTICIPANTS: Participants with mild-to-moderate AD (Mini-Mental State Examination score of 13-26) were recruited from December 1999 to November 2000 using clinic populations, referrals from community physicians, and local advertising. Stable use of cholinesterase inhibitors, estrogen, low-dose aspirin, and vitamin E was allowed. Participants with inflammatory diseases that might respond to the study medications were excluded. Of 474 participants screened, 351 were enrolled. INTERVENTIONS: Once-daily rofecoxib, 25 mg, or twice-daily naproxen sodium, 220 mg, or placebo. MAIN OUTCOME MEASURES: The primary outcome measure was the 1-year change in the Alzheimer Disease Assessment Scale-Cognitive (ADAS-Cog) subscale score. Secondary outcome measures included the Clinical Dementia Rating scale sum-of-boxes, the Neuropsychiatric Inventory, the Quality of Life-AD, and the time to attainment of significant end points (4-point decline from baseline ADAS-Cog score, 1-step worsening on the global Clinical Dementia Rating scale, 15-point decline on the ADCS activities of daily living inventory, institutionalization, or death). RESULTS: The 1-year mean (SD) change in ADAS-Cog scores in participants treated with naproxen (5.8 [8.0]) or rofecoxib (7.6 [7.7]) was not significantly different from the change in participants treated with placebo (5.7 [8.2]). Results of secondary analyses showed no consistent benefit of either treatment. Fatigue, dizziness, and hypertension were more commonly reported in the active drug groups, and more serious adverse events were found in the active treatment group than in the placebo group. CONCLUSION: The results of this study indicate that rofecoxib or low-dose naproxen does not slow cognitive decline in patients with mild-to-moderate AD. PMID- 12783913 TI - Paroxetine controlled release in the treatment of menopausal hot flashes: a randomized controlled trial. AB - CONTEXT: Standard therapy for hot flashes has been hormone replacement with estradiol or progestational agents, but recent data suggest that antidepressants inhibiting serotonin reuptake may also be effective. OBJECTIVE: To evaluate a selective serotonin reuptake inhibitor (paroxetine controlled release [CR]) in treating the vasomotor symptoms displayed by a general cross-section of menopausal women. DESIGN AND SETTING: Randomized, double-blind, placebo controlled, parallel group study conducted across 17 US sites, including urban, suburban, and rural clinics. PATIENTS: A total of 165 menopausal women aged 18 years or older experiencing at least 2 to 3 daily hot flashes and must have discontinued any hormone replacement therapy for at least 6 weeks. Women were excluded if they had any signs of active cancer or were undergoing chemotherapy or radiation therapy. INTERVENTION: After a 1-week placebo run-in phase, study participants were randomized to receive placebo or receive 12.5 mg/d or 25.0 mg/d of paroxetine CR (in a 1:1:1 ratio) for 6 weeks. MAIN OUTCOME MEASURES: Mean change from baseline to week 6 in the daily hot flash composite score (frequency x severity). RESULTS: Fifty-six participants were randomly assigned to receive placebo and 51 to receive 12.5 mg/d and 58 to receive 25.0 mg/d of paroxetine CR. The mean reductions in the hot flash frequency composite score from baseline to week 6 were statistically significantly greater for those receiving paroxetine CR than for those receiving placebo. By week 6, the mean daily hot flash frequency went from 7.1 to 3.8 (mean reduction, 3.3) for those in the 12.5-mg/d and from 6.4 to 3.2 (mean reduction, 3.2) for those in the 25-mg/d paroxetine CR groups and from 6.6 to 4.8 (mean reduction, 1.8) for those in the placebo group. Mean placebo-adjusted reduction in hot flash composite scores were -4.7 (95% confidence interval, - 8.1 to -1.3; P =.007) comparing 12.5-mg/d paroxetine CR with placebo; and -3.6 (95% confidence interval, -6.8 to -0.4; P =.03) comparing 25.0-mg/d paroxetine CR with placebo. This corresponded to median reductions of 62.2% for those in the 12.5-mg/d and 64.6% for those in the 25.0-mg/d paroxetine CR groups compared with 37.8% for those in the placebo group. CONCLUSION: Paroxetine CR may be an effective and acceptable alternative to hormone replacement and other therapies in treating menopausal hot flash symptoms. PMID- 12783914 TI - Belt-positioning booster seats and reduction in risk of injury among children in vehicle crashes. AB - CONTEXT: Although more than a dozen states have ratified laws that require booster seats for children older than 4 years, most states continue to have child restraint laws that only cover children through age 4 years. Lack of booster seat effectiveness data may be a barrier to passage of stronger child restraint laws. OBJECTIVES: To quantify the association of belt-positioning booster seats compared with seat belts alone and risk of injury among 4- to 7-year-old children and to assess patterns of injury among children in booster seats vs seat belts. DESIGN, SETTING, AND POPULATION: Cross-sectional study of children aged 4 to 7 years in crashes of insured vehicles in 15 states, with data collected via insurance claims records and a telephone survey. A probability sample of 3616 crashes involving 4243 children, weighted to represent 56 593 children in 48 257 crashes was collected between December 1, 1998, and May 31, 2002. MAIN OUTCOME MEASURE: Parent report of clinically significant injuries. RESULTS: Injuries occurred among 1.81% of all 4- to 7-year-olds, including 1.95% of those in seat belts and 0.77% of those in belt-positioning booster seats. The odds of injury, adjusting for child, driver, crash, and vehicle characteristics, were 59% lower for children aged 4 to 7 years in belt-positioning boosters than in seat belts (odds ratio, 0.41; 95% confidence interval, 0.20-0.86). Children in belt positioning booster seats had no injuries to the abdomen, neck/spine/back, or lower extremities, while children in seat belts alone had injuries to all body regions. CONCLUSION: Belt-positioning booster seats were associated with added safety benefits compared with seat belts to children through age 7 years, including reduction of injuries classically associated with improper seat belt fit in children. PMID- 12783915 TI - Length of stay in home care before and after the 1997 Balanced Budget Act. AB - CONTEXT: Prior to 1997, home health agencies (HHAs) were reimbursed on a fee-for service basis and had incentives to provide more services. The 1997 Balanced Budget Act (BBA) reduced payments for home care services to help control Medicare spending. OBJECTIVE: To examine the length of stay in home care before and after the 1997 BBA. DESIGN AND SETTING: Cross-sectional study of home care patients in the 1996 and 1998 National Home and Hospice Care Surveys, which surveyed 1053 HHAs in 1996 and 1088 HHAs in 1998. PATIENTS: Nationally representative random sample of home care patients with Medicare coverage in 1996 (4127 patients) and 1998 (4051 patients). MAIN OUTCOME MEASURE: Length of stay in home care (based on the number of days a patient was enrolled in home care services). RESULTS: From 1996 to 1998, unadjusted median length of stay decreased by 16 days for all home care patients (60-44 days, P =.002). The decrease affected for-profit HHAs more than not-for-profit HHAs (111-55 days [51% decrease, P =.002] vs 46-36 days [22% decrease, P =.042]). In a Cox proportional hazards model of time to discharge from home care, post-BBA year (1998) was associated with a shorter length of stay in home care (adjusted hazard ratio [aHR] for home care discharge, 1.39 [95% confidence interval [CI], 1.19-1.61]), and for-profit status was associated with a longer length of stay in home care (aHR, 0.82 [95% CI, 0.71-0.94]) after adjusting for patient demographics, diagnoses, and functional status. CONCLUSION: After the 1997 BBA, length of stay in home care decreased among Medicare patients, particularly among those receiving care from for-profit HHAs. PMID- 12783916 TI - Changes in rates of autopsy-detected diagnostic errors over time: a systematic review. AB - CONTEXT: Substantial discrepanies exist between clinical diagnoses and findings at autopsy. Autopsy may be used as a tool for quality management to analyze diagnostic discrepanies. OBJECTIVE: To determine the rate at which autopsies detect important, clinically missed diagnoses, and the extent to which this rate has changed over time. DATA SOURCES: A systematic literature search for English language articles available on MEDLINE from 1966 to April 2002, using the search terms autopsy, postmortem changes, post-mortem, postmortem, necropsy, and posthumous, identified 45 studies reporting 53 distinct autopsy series meeting prospectively defined criteria. Reference lists were reviewed to identify additional studies, and the final bibliography was distributed to experts in the field to identify missing or unpublished studies. STUDY SELECTION: Included studies reported clinically missed diagnoses involving a primary cause of death (major errors), with the most serious being those likely to have affected patient outcome (class I errors). DATA EXTRACTION: Logistic regression was performed using data from 53 distinct autopsy series over a 40-year period and adjusting for the effects of changes in autopsy rates, country, case mix (general autopsies; adult medical; adult intensive care; adult or pediatric surgery; general pediatrics or pediatric inpatients; neonatal or pediatric intensive care; and other autopsy), and important methodological features of the primary studies. DATA SYNTHESIS: Of 53 autopsy series identified, 42 reported major errors and 37 reported class I errors. Twenty-six autopsy series reported both major and class I error rates. The median error rate was 23.5% (range, 4.1%-49.8%) for major errors and 9.0% (range, 0%-20.7%) for class I errors. Analyses of diagnostic error rates adjusting for the effects of case mix, country, and autopsy rate yielded relative decreases per decade of 19.4% (95% confidence interval [CI], 1.8%-33.8%) for major errors and 33.4% (95% [CI], 8.4%-51.6%) for class I errors. Despite these decreases, we estimated that a contemporary US institution (based on autopsy rates ranging from 100% [the extrapolated extreme at which clinical selection is eliminated] to 5% [roughly the national average]), could observe a major error rate from 8.4% to 24.4% and a class I error rate from 4.1% to 6.7%. CONCLUSION: The possibility that a given autopsy will reveal important unsuspected diagnoses has decreased over time, but remains sufficiently high that encouraging ongoing use of the autopsy appears warranted. PMID- 12783917 TI - Gout. PMID- 12783918 TI - MRI for regional back pain: need for less imaging, better understanding. PMID- 12783919 TI - Nonsteroidal anti-inflammatory drugs and Alzheimer disease: what's next? PMID- 12783920 TI - JAMA patient page. Autopsy. PMID- 12783922 TI - NRAS hypermutability in familial melanoma with CDKN2A mutations--cause and effect? PMID- 12783923 TI - Why it hurts: researchers seek mechanisms of cancer pain. PMID- 12783924 TI - Researchers push for publication, registration of all clinical trials. PMID- 12783926 TI - Work on breast cancer stem cells raises questions about treatment strategies. PMID- 12783927 TI - Cancer rates change slightly with census data correction. PMID- 12783921 TI - Is low-dose tamoxifen useful for the treatment and prevention of breast cancer? PMID- 12783928 TI - Stat bite: Trends in cancer incidence rates by primary cancer sites, 1992-2000. PMID- 12783932 TI - A randomized trial of low-dose tamoxifen on breast cancer proliferation and blood estrogenic biomarkers. AB - BACKGROUND: Tamoxifen reduces the risk of breast cancer in women at high risk for the disease but increases the risk for endometrial tumors and venous thromboembolisms, possibly in a dose-dependent fashion. We compared the effects of tamoxifen at 1 mg/day and 5 mg/day with those of the standard dose of 20 mg/day on breast cancer proliferation using a surrogate endpoint marker (Ki-67 expression) and blood biomarkers associated with breast cancer, cardiovascular disease, and bone fracture risk. METHODS: We randomly assigned 120 women with estrogen receptor (ER)-positive breast cancer to tamoxifen at 1, 5, or 20 mg/day for 4 weeks. Expression of the tumor proliferation marker Ki-67 and of biomarkers of breast cancer (insulin-like growth factor-I, sex hormone-binding globulin), cardiovascular disease (cholesterol, triglycerides, ultrasensitive C-reactive protein, fibrinogen, antithrombin-III), and bone fracture (type I collagen C telopeptide) risk were determined before (baseline) and after treatment. All levels were compared with those in two nonrandomized control groups (34 women with ER-negative breast cancer and 29 additional women with ER-positive breast cancer). Data were analyzed by analysis of covariance. All statistical tests were two-sided. RESULTS: Expression of Ki-67 decreased in all three tamoxifen groups, with no difference in the magnitude of reduction among groups (P =.81). Relative to baseline, Ki-67 expression decreased by a median of 15.0% (95% confidence interval = 0.0% to 24.1%) among the tamoxifen groups but increased by 12.8% (95% confidence interval = 0.0% to 19.6%) among the nonrandomized control groups. Several blood biomarkers showed dose-response relationships with tamoxifen, including decreased insulin-like growth factor-I, increased sex hormone-binding globulin, and decreased low-density lipoprotein-cholesterol, ultrasensitive C reactive protein, fibrinogen, and antithrombin-III levels. CONCLUSIONS: The effects on Ki-67 expression of lower doses of tamoxifen were comparable to those achieved with the standard dose, although the effects on blood biomarkers were variable. The effects of lower doses of tamoxifen should be assessed further in randomized trials. PMID- 12783933 TI - Frequency of UV-inducible NRAS mutations in melanomas of patients with germline CDKN2A mutations. AB - BACKGROUND: Germline alterations in cyclin-dependent kinase inhibitor 2A (CDKN2A) are important genetic factors in familial predisposition to melanoma. Activating mutations of the NRAS proto-oncogene are among the most common somatic genetic alterations in cutaneous malignant melanomas. We investigated the occurrence of NRAS mutations in melanomas and dysplastic nevi in individuals with germline CDKN2A mutations. METHODS: Genomic DNA was extracted from 39 biopsy samples (including primary melanomas, metastatic melanomas, and dysplastic nevi) from 25 patients in six Swedish families with a hereditary predisposition to melanoma who carried germline CDKN2A mutations. DNA was also extracted from 10 biopsy samples from patients with sporadic melanomas. NRAS was analyzed using polymerase chain reaction, single-strand conformation polymorphism analysis, and nucleotide sequence analysis. Differences in NRAS mutation frequency between hereditary and sporadic melanomas were analyzed by the chi-square test. All statistical tests were two-sided. RESULTS: Activating mutations in NRAS codon 61, all of which were either CAA(Gln)-AAA(Lys) or CAA(Gln)-CGA(Arg) mutations, were found in 95% (20/21) of primary hereditary melanomas but in only 10% (1/10) of sporadic melanomas (P<.001). Multiple activating NRAS mutations were detected in tumor cells from different regions of individual primary melanomas in nine patients. Activating mutations that were detected in the primary melanomas of these patients were also retained in their metastases. NRAS mutations at sites other than codon 61 were also present in the primary melanomas, indicating genetic instability of this locus. NRAS codon 61 mutations were also detected in dysplastic nevi and in an in situ melanoma, suggesting a role for such mutations during early melanoma development. CONCLUSIONS: The high frequency of NRAS codon 61 mutations detected in these hereditary melanomas may be the result of a hypermutability phenotype associated with a hereditary predisposition for melanoma development in patients with germline CDKN2A mutations. PMID- 12783934 TI - Personality and the risk of cancer. AB - BACKGROUND: The role of personality in the causation of cancer has been controversial. We examined this question in a large, prospective study. METHODS: From June through August 1990, 30 277 residents of Miyagi Prefecture in northern Japan completed a Japanese version of the short form of the Eysenck Personality Questionnaire-Revised and a questionnaire on various health habits. There were 671 prevalent cases of cancer at baseline, and 986 incident cases of cancer were identified during 7 years of follow-up, through December 1997. We used Cox proportional hazards regression to estimate the relative risk (RR) of incident cancer (total, stomach, colorectal, breast, and lung) according to four levels of each of four personality subscales (extraversion, neuroticism, psychoticism, and lie), with adjustment for sex, age, education, smoking, alcohol use, body mass index, and family history of cancer. Statistical tests were two-sided. RESULTS: Multivariable RRs of total cancer for individuals in the highest level of each personality subscale as compared with those in the lowest were 0.9 for extraversion (95% confidence interval [CI] = 0.7 to 1.1; P(trend) =.32), 1.1 for psychoticism (95% CI = 0.9 to 1.3; P(trend) =.96), 0.9 for lie (95% CI = 0.7 to 1.0; P(trend) =.19), and 1.2 for neuroticism (95% CI = 1.0 to 1.4; P(trend) =.06). There were no associations between any personality subscale and risk of specific cancers. Neuroticism showed statistically significant positive, linear associations with prevalent cancer at baseline (P(trend)<.001) and with the 320 incident cancer cases diagnosed within the first 3 years of follow-up (P(trend) =.03); however, it showed no association with the 666 cases diagnosed during the fourth through the seventh years of follow-up (P(trend) =.43). CONCLUSION: Our data do not support the hypothesis that personality is a risk factor for cancer incidence. The association between neuroticism and prevalent cancer may be a consequence, rather than a cause, of cancer diagnosis or symptoms. PMID- 12783935 TI - Melanocytic nevi, solar keratoses, and divergent pathways to cutaneous melanoma. AB - BACKGROUND: Some melanomas form on sun-exposed body sites, whereas others do not. We previously proposed that melanomas at different body sites arise through different pathways that have different associations with melanocytic nevi and solar keratoses. We tested this hypothesis in a case-case comparative study of melanoma patients in Queensland, Australia. METHODS: We randomly selected patients from among three prespecified groups reported to the population-based Queensland Cancer Registry: those with superficial spreading or nodular melanomas of the trunk (n = 154, the reference group), those with such melanomas of the head and neck (n = 77, the main comparison group), and those with lentigo maligna melanoma (LMM) (n = 75, the chronic sun-exposed group). Each participant completed a questionnaire, and a research nurse counted melanocytic nevi and solar keratoses. We calculated exposure odds ratios (ORs) and 95% confidence intervals (CIs) to quantify the association between factors of interest and each melanoma group. RESULTS: Patients with head and neck melanomas, compared with patients with melanomas of the trunk, were statistically significantly less likely to have more than 60 nevi (OR = 0.34, 95% CI = 0.15 to 0.79) but were statistically significantly more likely to have more than 20 solar keratoses (OR = 3.61, 95% CI = 1.42 to 9.17) and also tended to have a past history of excised solar skin lesions (OR = 1.87, 95% CI = 0.89 to 3.92). Patients with LMM were also less likely than patients with truncal melanomas to have more than 60 nevi (OR = 0.32, 95% CI = 0.14 to 0.75) and tended toward more solar keratoses (OR = 2.14, 95% CI = 0.88 to 5.16). CONCLUSIONS: Prevalences of nevi and solar keratoses differ markedly between patients with head and neck melanomas or LMM and patients with melanomas of the trunk. Cutaneous melanomas may arise through two pathways, one associated with melanocyte proliferation and the other with chronic exposure to sunlight. PMID- 12783936 TI - New paraoxonase 1 polymorphism I102V and the risk of prostate cancer in Finnish men. AB - BACKGROUND: Human serum paraoxonase eliminates carcinogenic lipid-soluble radicals. Because expression of the main human paraoxonase gene PON1 varies widely in humans, certain PON1 polymorphisms might be associated with increased risks of cancer. We sought new functional mutations in PON1 and determined whether known or new PON1 mutations were associated with the risk for prostate cancer in a prospective, random, population-based sample of Finnish men and in a case-control study. METHODS: Serum paraoxonase activity was measured in 835 healthy men in the Kuopio Ischaemic Heart Disease Risk Factor Study. PON1 mutations were identified by hierarchical phenotype-targeted sequencing in DNAs from the 100 men with the lowest paraoxonase activity in this cohort, and 1595 men in the cohort were genotyped for PON1 mutations by restriction fragment length polymorphism. Multivariable analysis was used to investigate the association of known and new PON1 mutations with incident prostate cancer in 1569 cancer-free men in the cohort followed for 9-14 years. In a case-control study of Finnish men, the association of prostate cancer with the PON1 mutation identified in the cohort study was investigated in 69 case patients with familial prostate cancer and 69 unmatched healthy control subjects. RESULTS: We identified a new single-nucleotide PON1 polymorphism associated with decreased serum paraoxonase activity that caused an isoleucine-->valine change at codon 102 in exon 4 (I102V). Of the 1569 men cancer-free at baseline, 56 (3.6%) were carriers of the I102V mutation. After adjusting for age and cholesterol-lowering medications, the relative risk for developing prostate cancer during follow-up was 6.3 (95% confidence interval [CI] = 2.1 to 19.2) among 102V allele carriers compared with noncarriers. Other PON1 alleles were not statistically significantly associated with prostate cancer. In the case-control study, patients with familial prostate cancer were more likely to be carriers of the PON1 I102V mutation than control subjects (odds ratio = 4.3, 95% CI = 0.9 to 21.5). CONCLUSION: The PON1 102V allele appears to be associated with an increased risk for prostate cancer. PMID- 12783937 TI - ELAC2/HPC2 polymorphisms, prostate-specific antigen levels, and prostate cancer. AB - BACKGROUND: The ELAC2 gene has been proposed to be a prostate cancer susceptibility gene and is being referred to as HPC2, in part because three case control studies suggested that two common polymorphisms (Ser217Leu and Ala541Thr) are associated with risk. However, four subsequent larger studies have not confirmed this association. In five of the seven total studies, subject selection was influenced by prostate-specific antigen (PSA) levels. We examined the association and possible effect of subject selection in a larger study and a meta analysis. METHODS: In a population-based study in Australia, 825 case patients and 732 control subjects were genotyped for the Ser217Leu and Ala541Thr polymorphisms of ELAC2. Odds ratios (ORs) for prostate cancer were estimated by unconditional logistic and polytomous regression. A meta-analysis was conducted combining our data with those from seven published studies. The association of genotype with the logarithm of plasma PSA levels in control subjects was analyzed by linear regression. RESULTS: The ORs for prostate cancer were 0.74 (95% confidence interval [CI] = 0.50 to 1.09) for Leu217 homozygotes and 1.01 (95% CI = 0.68 to 1.50) for Thr541 heterozygotes and homozygotes compared with Ser217 and Ala541 homozygotes, respectively. ORs were not changed by excluding control subjects with elevated PSA levels. Among control subjects, there were no statistically significant associations between genotype frequencies and PSA level for either polymorphism (both P>.4). The meta-analysis gave pooled OR estimates of 1.04 (95% CI = 0.85 to 1.26) for Leu217 homozygotes and 1.18 (OR = 0.98 to 1.42) for Thr541 homozygotes and heterozygotes. CONCLUSION: There is no evidence that either ELAC2 polymorphism is associated with prostate cancer or PSA level. PMID- 12783938 TI - Night-shift work and risk of colorectal cancer in the nurses' health study. AB - Exposure to light at night suppresses the physiologic production of melatonin, a hormone that has antiproliferative effects on intestinal cancers. Although observational studies have associated night-shift work with an increased risk of breast cancer, the effect of night-shift work on the risk of other cancers is not known. We prospectively examined the relationship between working rotating night shifts and the risk of colorectal cancers among female participants in the Nurses' Health Study. We documented 602 incident cases of colorectal cancer among 78 586 women who were followed up from 1988 through 1998. Compared with women who never worked rotating night shifts, women who worked 1-14 years or 15 years or more on rotating night shifts had multivariate relative risks of colorectal cancer of 1.00 (95% confidence interval [CI] = 0.84 to 1.19) and 1.35 (95% CI = 1.03 to 1.77), respectively (P(trend) =.04). These data suggest that working a rotating night shift at least three nights per month for 15 or more years may increase the risk of colorectal cancer in women. PMID- 12783939 TI - Re: Association between hemochromatosis (HFE) gene mutation carrier status and the risk of colon cancer. PMID- 12783940 TI - Re: Microarray studies challenge theories of metastasis. PMID- 12783941 TI - Re: Risk of perforation after colonoscopy and sigmoidoscopy: a population-based study. PMID- 12783943 TI - Successful treatment of thrombocytopenia due to marrow metastases of breast cancer with weekly docetaxel. PMID- 12783945 TI - Branching out: a new idea for dendritic function. Focus on "Coincidence detection in pyramidal neurons is tuned by their dendritic branching pattern". PMID- 12783946 TI - Control strategies correcting inaccurately programmed fingertip forces: model predictions derived from human behavior. AB - When picking up a familiar object between the index finger and the thumb, the motor commands are predetermined by the CNS to correspond to the frictional demand of the finger-object contact area. If the friction is less than expected, the object will start to slip out of the hand, giving rise to unexpected sensory information. Here we study the correction strategies of the motor system in response to an unexpected frictional demand. The motor commands to the mononeuron pool are estimated by a novel technique combining behavioral recordings and neuromuscular modelling. We first propose a mathematical model incorporating muscles, hand mechanics, and the action of lifting an object. A simple control system sends motor commands to and receives sensory signals from the model. We identify three factors influencing the efficiency of the correction: the time development of the motor command, the delay between the onset of the grip and load forces (GF-LF-delay), and how fast the lift is performed. A sensitivity analysis describes how these factors affect the ability to prevent or stop slipping and suggests an efficient control strategy that prepares and corrects for an altered frictional condition. We then analyzed fingertip grip and load forces (GF and LF) and position data from 200 lifts made by five healthy subjects. The friction was occasionally reduced, forcing an increase of the GF to prevent the object being dropped. The data were then analyzed by feeding it through the inverted model. This provided an estimate of the motor commands to the motoneuron pool. As suggested by the sensitivity analysis the GF-LF-delay was indeed used by the subjects to prevent slip. In agreement with recordings from neurons in the primary motor cortex of the monkey, a sharp burst in the estimated GF motor command (NGF) efficiently arrested any slip. The estimated motor commands indicate a control system that uses a small set of corrective commands, which together with the GF-LF-delay form efficient correction strategies. The selection of a strategy depends on the amount of tactile information reporting unexpected friction and how long it takes to arrive. We believe that this technique of estimating the motor commands behind the fingertip forces during a precision grip lift can provide a powerful tool for the investigation of the central control of the motor system. PMID- 12783947 TI - Cancelling of pursuit and saccadic eye movements in humans and monkeys. AB - The countermanding paradigm provides a useful tool for examining the mechanisms responsible for cancelling eye movements. The key feature of this paradigm is that, on a minority of trials, a stop signal is introduced some time after the appearance of the target, indicating that the subject should cancel the incipient eye movement. If the delay in giving the stop signal is too long, subjects fail to cancel the eye movement to the target stimulus. By modeling this performance as a race between a go process triggered by the appearance of the target and a stop process triggered by the appearance of the stop signal, it is possible to estimate the processing interval associated with cancelling the movement. We have now used this paradigm to analyze the cancelling of pursuit and saccades. For pursuit, we obtained consistent estimates of the stop process regardless of our technique or assumptions--it took 50-60 ms to cancel pursuit in both humans and monkeys. For saccades, we found different values depending on our assumptions. When we assumed that saccade preparation was under inhibitory control up until movement onset, we found that saccades took longer to cancel (humans: approximately 110, monkeys: approximately 80 ms) than pursuit. However, when we assumed that saccade preparation includes a final "ballistic" interval not under inhibitory control, we found that the same rapid stop process that accounted for our pursuit results could also account for the cancelling of saccades. We favor this second interpretation because cancelling pursuit or saccades amounts to maintaining a state of fixation, and it is more parsimonious to assume that this involves a single inhibitory process associated with the fixation system, rather than two separate inhibitory processes depending on which type of eye movement will not be made. From our behavioral data, we estimate that this ballistic interval has a duration of 9-25 ms in monkeys, consistent with the known physiology of the final motor pathways for saccades, although we obtained longer values in humans (28-60 ms). Finally, we examined the effect of trial sequence during the countermanding task and found that pursuit and saccade latencies tended to be longer if the previous trial contained a stop signal than if it did not; these increases occurred regardless of whether the preceding trial was associated with the same or different type of eye movement. Together, these results suggest that a common inhibitory mechanism regulates the initiation of pursuit and saccades. PMID- 12783948 TI - Caffeine-mediated presynaptic long-term potentiation in hippocampal CA1 pyramidal neurons. AB - We report a new form of long-term potentiation (LTP) in Schaffer collateral (SC) CA1 pyramidal neuron synapses that originates presynaptically and does not require N-methyl-d-aspartate (NMDA) receptor activation nor increases in postsynaptic-free Ca2+. Using rat hippocampal slices, application of a brief "pulse" of caffeine in the bath evoked a nondecremental LTP (CAFLTP) of SC excitatory postsynaptic currents. An increased probability of transmitter release paralleled the CAFLTP, suggesting that it originated presynaptically. The P1 adenosine receptor antagonist 8-cyclopentyltheophylline and the P2 purinoreceptor antagonists suramin and piridoxal-5'-phosphate-azophenyl 2',4'-disulphonate blocked the CAFLTP. Inhibition of Ca2+ release from caffeine/ryanodine stores by bath-applied ryanodine inhibited the CAFLTP, but ryanodine in the pipette solution was ineffective, suggesting a presynaptic effect of ryanodine. Previous induction of the "classical" LTP did not prevent the CAFLTP, suggesting that the LTP and the CAFLTP have different underlying cellular mechanisms. The CAFLTP is insensitive to the block of NMDA receptors by 2-amino-5-phosphonopentanoic acid and to Ca2+ chelation with intracellular 1,2-bis (2-aminophenoxy) ethane N,N,N',N'-tetraacetic acid, indicating that neither postsynaptic NMDA receptors nor increases in cytosolic-free Ca2+ participate in the CAFLTP. We conclude that the CAFLTP requires the interaction of caffeine with presynaptic P1, P2 purinoreceptors, and ryanodine receptors and is caused by an increased probability of glutamate release at SC terminals. PMID- 12783949 TI - Altered excitability of intestinal neurons in primary culture caused by acute oxidative stress. AB - Neurons were isolated from the intestine of guinea pigs and grown in primary culture for < or =15 days. Using conventional whole cell recording techniques, we demonstrated that the majority of neurons express a prolonged poststimulus afterhyperpolarization (slow AHP). These neurons also had large-amplitude (approximately 100 mV), broad-duration (approximately 2 ms) action potentials and generated a hyperpolarization activated inward current (Ih). Application of H2O2 (0.22-8.8 mM) hyperpolarized these neurons but not those lacking slow AHPs. The H2O2-induced hyperpolarization was followed by irreversible depolarization at higher concentrations (more than approximately 1 mM) of H2O2 while it was maintained after washout of submillimolar H2O2. The ionic mechanisms underlying the hyperpolarization included the suppression of Ih and the activation of an inwardly rectifying outward current, which was blocked by glybenclamide (25-50 microM) and TEA (30 mM). In addition, H2O2 suppressed the slow AHP and its underlying current. Internal perfusion of catalase and glutathione opposed the H2O2-mediated decrease in IsAHP. Our results indicate that acute oxidative stress has neuron- and conductance-specific actions in intestinal neurons that may underlie pathophysiological conditions. PMID- 12783950 TI - Dipole source analysis of laser-evoked subdural potentials recorded from parasylvian cortex in humans. AB - The location of the human nociceptive area(s) near the Sylvian fissure is still controversial in spite of evidence from imaging and evoked potential studies that noxious heat stimuli activate somatosensory areas in that region. Some studies have suggested the secondary somatosensory cortex (SII) on the upper bank of the Sylvian fissure posterior to the central sulcus, others the anterior insula or parietal area 7b. In this study, we applied dipole source analysis techniques to laser-evoked potentials (LEPs) that were recorded from subdural grid electrodes in three patients. As a functional marker, auditory-evoked potentials (AEPs) with a generator on the opposite bank of the Sylvian fissure were recorded from the same electrodes. The LEP global field power (GFP), a measure of spatial variance, showed a first peak at about 150 ms latency, corresponding to the latency of the N1 recorded from the scalp. In contrast to scalp recordings, the amplitude of the first GFP peak recorded from the grid was larger than the second peak (P2). This finding suggests that the generator of N1, but not that of later LEP components, was close to the subdural grids. When a regional source was fitted to the first GFP peak, its location was within the frontoparietal operculum in all patients. On average, the LEP source was 13 mm anterior, 6 mm superior, and 2 mm medial of the AEP source. This relative location also suggests a source within the frontoparietal operculum overlying the insula. At the latency of the first GFP peak, source orientation pointed inward, suggesting a generator within the inner vertical surface of the operculum. Somatotopy was assessed in one patient and was consistent with that of the projection area of the presumed nociceptive thalamic nucleus posterior part of the ventromedial nucleus, but differed from that of SII. These findings suggest that the nociceptive area in human parasylvian cortex that is activated most rapidly by noxious heat pulses may be separate from the tactile SII area. PMID- 12783951 TI - Differential expression of three distinct potassium currents in the ventral cochlear nucleus. AB - In the ventral cochlear nucleus (VCN), neurons transform information from auditory nerve fibers into a set of parallel ascending pathways, each emphasizing different aspects of the acoustic environment. Previous studies have shown that VCN neurons differ in their intrinsic electrical properties, including the K+ currents they express. In this study, we examine these K+ currents in more detail using whole cell voltage-clamp techniques on isolated VCN cells from adult guinea pigs at 22 degrees C. Our results show a differential expression of three distinct K+ currents. Whereas some VCN cells express only a high-threshold delayed-rectifier-like current (IHT), others express IHT in combination with a fast inactivating current (IA) and/or a slow-inactivating low-threshold current (ILT). IHT, ILT, and IA, were partially blocked by 1 mM 4-aminopyridine. In contrast, only ILT was blocked by 10-100 nM dendrotoxin-I. A surprising finding was the wide range of levels of ILT, suggesting ILT is expressed as a continuum across cell types rather than modally in a particular cell type. IA, on the other hand, appears to be expressed only in cells that show little or no ILT, the Type I cells. Boltzmann analysis shows IHT activates with 164 +/- 12 (SE) nS peak conductance, -14.3 +/- 0.7 mV half-activation, and 7.0 +/- 0.5 mV slope factor. Similar analysis shows ILT activates with 171 +/- 22 nS peak conductance, -47.4 +/- 1.0 mV half-activation, and 5.8 +/- 0.3 mV slope factor. PMID- 12783952 TI - Kinetic analyses of three distinct potassium conductances in ventral cochlear nucleus neurons. AB - Neurons in the ventral cochlear nucleus (VCN) express three distinct K+ currents that differ in their voltage and time dependence, and in their inactivation behavior. In the present study, we quantitatively analyze the voltage-dependent kinetics of these three currents to gain further insight into how they regulate the discharge patterns of VCN neurons and to provide supporting data for the identification of their channel components. We find the transient A-type K+ current (IA) exhibits fourth-order activation kinetics (a4), and inactivates with one or two time constants. A second inactivation rate (leading to an a4bc kinetic description) is required to explain its recovery from inactivation. The dendrotoxin-sensitive low-threshold K+ current (ILT) also activates with fourth order kinetics (w4) but shows slower, incomplete inactivation. The high-threshold K+ current (IHT) appears to consist of two kinetically distinct components (n2 + p). The first component activates approximately 10 mV positive to the second and has second-order kinetics. The second component activates with first-order kinetics. These two components also contribute to two kinetically distinct currents upon deactivation. The kinetic behavior of IHT was indistinguishable amongst cell types, suggesting the current is mediated by the same K+ channels amongst VCN neurons. Together these results provide a basis for more realistic modeling of VCN neurons, and provide clues regarding the molecular basis of the three K+ currents. PMID- 12783953 TI - The roles potassium currents play in regulating the electrical activity of ventral cochlear nucleus neurons. AB - Using kinetic data from three different K+ currents in acutely isolated neurons, a single electrical compartment representing the soma of a ventral cochlear nucleus (VCN) neuron was created. The K+ currents include a fast transient current (IA), a slow-inactivating low-threshold current (ILT), and a noninactivating high-threshold current (IHT). The model also includes a fast inactivating Na+ current, a hyperpolarization-activated cation current (Ih), and 1-50 auditory nerve synapses. With this model, the role IA, ILT, and IHT play in shaping the discharge patterns of VCN cells is explored. Simulation results indicate that IHT mainly functions to repolarize the membrane during an action potential, and IA functions to modulate the rate of repetitive firing. ILT is found to be responsible for the phasic discharge pattern observed in Type II cells (bushy cells). However, by adjusting the strength of ILT, both phasic and regular discharge patterns are observed, demonstrating that a critical level of ILT is necessary to produce the Type II response. Simulated Type II cells have a significantly faster membrane time constant in comparison to Type I cells (stellate cells) and are therefore better suited to preserve temporal information in their auditory nerve inputs by acting as precise coincidence detectors and having a short refractory period. Finally, we demonstrate that modulation of Ih, which changes the resting membrane potential, is a more effective means of modulating the activation level of ILT than simply modulating ILT itself. This result may explain why ILT and Ih are often coexpressed throughout the nervous system. PMID- 12783954 TI - Neuronal responses in cat primary auditory cortex to electrical cochlear stimulation: IV. Activation pattern for sinusoidal stimulation. AB - Patterns of threshold distributions for single-cycle sinusoidal electrical stimulation and single pulse electrical stimulation were compared in primary auditory cortex of the adult cat. Furthermore, the effects of auditory deprivation on these distributions were evaluated and compared across three groups of adult cats. Threshold distributions for single and multiple unit responses from the middle cortical layers were obtained on the ectosylvian gyrus in an acutely implanted animal; 2 wk after deafening and implantation (short-term group); and neonatally deafened animals implanted following 2-5 yr of deafness (long-term group). For all three cases, we observed similar patterns of circumscribed regions of low response thresholds in the region of primary auditory cortex (AI). A dorsal and a ventral region of low response thresholds were found separated by a narrow, anterior-posterior strip of elevated thresholds. The ventral low-threshold regions in the short-term group were cochleotopically arranged. By contrast, the dorsal region in the short-term animals and both low-threshold regions in long-term deafened animals maintained only weak cochleotopicity. Analysis of the spatial extent of the low-threshold regions revealed that the activated area for sinusoidal stimulation was smaller and more circumscribed than for pulsatile stimulation for both dorsal and ventral AI. The width of the high-threshold ridge that separated the dorsal and ventral low-threshold regions was greater for sinusoidal stimulation. Sinusoidal and pulsatile threshold behavior differed significantly for electrode configurations with low and high minimum thresholds. Differences in threshold behavior and cortical response distributions between the sinusoidal and pulsatile stimulation suggest that stimulus shape plays a significant role in the activation of cortical activity. Differences in the activation pattern for short-term and long term deafness reflect deafness-induced reorganizational changes based on factors such as differences in excitatory and inhibitory balance that are affected by the stimulation parameters. PMID- 12783955 TI - Reorganization of remote cortical regions after ischemic brain injury: a potential substrate for stroke recovery. AB - Although recent neurological research has shed light on the brain's mechanisms of self-repair after stroke, the role that intact tissue plays in recovery is still obscure. To explore these mechanisms further, we used microelectrode stimulation techniques to examine functional remodeling in cerebral cortex after an ischemic infarct in the hand representation of primary motor cortex in five adult squirrel monkeys. Hand preference and the motor skill of both hands were assessed periodically on a pellet retrieval task for 3 mo postinfarct. Initial postinfarct motor impairment of the contralateral hand was evident in each animal, followed by a gradual improvement in performance over 1-3 mo. Intracortical microstimulation mapping at 12 wk after infarct revealed substantial enlargements of the hand representation in a remote cortical area, the ventral premotor cortex. Increases ranged from 7.2 to 53.8% relative to the preinfarct ventral premotor hand area, with a mean increase of 36.0 +/- 20.8%. This enlargement was proportional to the amount of hand representation destroyed in primary motor cortex. That is, greater sparing of the M1 hand area resulted in less expansion of the ventral premotor cortex hand area. These results suggest that neurophysiologic reorganization of remote cortical areas occurs in response to cortical injury and that the greater the damage to reciprocal intracortical pathways, the greater the plasticity in intact areas. Reorganization in intact tissue may provide a neural substrate for adaptive motor behavior and play a critical role in postinjury recovery of function. PMID- 12783956 TI - Ca2+ clearance at growth cones produced by crayfish motor axons in an explant culture. AB - Intracellular free Ca2+ concentration ([Ca2+]i) plays an important role in the regulation of growth cone (GC) motility; however, the mechanisms responsible for clearing Ca2+ from GCs have not been examined. We studied the Ca2+-clearance mechanisms in GCs produced by crayfish tonic and phasic motor axons by measuring the decay of [Ca2+]i after a high [K+] depolarizing pulse using fura-2AM. Tonic motor axons regenerating in explant cultures develop GCs with more rapid Ca2+ clearance than GCs from phasic axons. When Na/Ca exchange was blocked by replacing external Na+ with N-methyl-d-glucamine (NMG), [Ca2+]i decay was delayed in both tonic and phasic GCs. Tonic GCs appear to have higher Na/Ca exchange activity than phasic ones since reversal of Na/Ca exchange by lowering external Na+ caused a greater increase in [Ca2+]i for tonic than phasic GCs. Application of the mitochondrial inhibitors, Antimycin A1 (1 microM) and CCCP (10 microM), demonstrated that mitochondrial Ca2+ uptake/release was more prominent in phasic than tonic GCs. When both Na/Ca exchange and mitochondria were inhibited, the plasma membrane Ca2+ ATPase was effective in extruding Ca2+ from tonic, but not phasic GCs. We conclude that Na/Ca exchange plays a prominent role in extruding large Ca2+ loads from both tonic and phasic GCs. High Na/Ca exchange activity in tonic GCs contributes to the rapid decay of [Ca2+]i in these GCs; low rates of Ca2+ extrusion plus the release of Ca2+ from mitochondria prolongs the decay of [Ca2+]i in the phasic GCs. PMID- 12783957 TI - TNP-ATP-resistant P2X ionic current on the central terminals and somata of rat primary sensory neurons. AB - P2X receptors have been suggested to be expressed on the central terminals of A delta-afferent fibers innervating dorsal horn lamina V and play a role in modulating sensory synaptic transmission. These P2X receptors have been widely thought to be P2X2+3 receptors. However, we have recently found that P2X receptor mediated modulation of sensory transmission in lamina V is not inhibited by trinitrophenyl-adenosine triphosphate (TNP-ATP), a potent antagonist of P2X1, P2X3 homomers, and P2X2+3 heteromers. To provide direct evidence for the presence of TNP-ATP-resistant P2X receptors on primary afferent fibers, we examined alpha,beta-methylene-ATP (alpha beta meATP)-evoked currents and their sensitivity to TNP-ATP in rat dorsal root ganglion (DRG) neurons. alpha beta meATP evoked fast currents, slow currents, and mixed currents that contained both fast and slow current-components. Fast currents and fast current components in the mixed currents were both completely inhibited by 0.1 microM TNP-ATP (n = 14). Both slow currents and slow-current components in the mixed currents showed broad spectrum of sensitivity to 1 microM TNP-ATP, ranging from complete block (TNP-ATP sensitive) to little block (TNP-ATP-resistant). TNP-ATP-resistant currents evoked by 10 microM alpha beta meATP could be largely inhibited by 10 microM iso pyridoxalphosphate-6-azophenyl-2',4'-disulphonic acid. Cells with P2X currents that were highly resistant to TNP-ATP were found to be insensitive to capsaicin. These results suggest that TNP-ATP-resistant P2X receptor subtypes are expressed on capsaicin-insensitive A delta-afferent fibers and play a role in modulating sensory transmission to lamina V neurons. PMID- 12783958 TI - Distinct roles of P2X receptors in modulating glutamate release at different primary sensory synapses in rat spinal cord. AB - Using spinal cord slice preparations and patch-clamp recordings in lamina II and lamina V regions, we tested a hypothesis that P2X receptor subtypes differentially modulate glutamate release from primary afferent terminals innervating different sensory regions. We found that activation of P2X receptors by alpha,beta-methylene-ATP increased glutamate release onto >80% of DH neurons in both lamina regions. However, two distinct types of modulation, a transient and a long-lasting enhancement of glutamate release were observed. In lamina II recordings, >70% of the modulation was transient. In contrast, P2X receptor mediated modulation was always long-lasting in lamina V. Pharmacologically, both transient and long-lasting types of modulation were blocked by 10 microM pyridxal phosphate-6-azophenyl-2',4'-disulphonic acid tetrasodium, a broad-spectrum P2X receptor antagonist. Transient modulation was not observed in the presence of 1 microM trinitrophenyl-ATP (TNP-ATP), a subtype-selective P2X receptor antagonist, suggesting that homomeric P2X3 receptors may be involved in the transient modulation in lamina II. The long-lasting modulation remained in the presence of 1 microM TNP-ATP. Selective removal of P2X3-expressing afferent terminals by the targeting toxin saporin-conjugated isolectin B4 or surgical removal of superficial DH did not affect P2X receptor-mediated long-lasting modulation in lamina V. Taken together, these results suggest that P2X receptor subtypes play distinct roles in sensory processing in functionally different sensory regions. PMID- 12783959 TI - Anterior inferotemporal neurons of monkeys engaged in object recognition can be highly sensitive to object retinal position. AB - Visual object recognition is computationally difficult because changes in an object's position, distance, pose, or setting may cause it to produce a different retinal image on each encounter. To robustly recognize objects, the primate brain must have mechanisms to compensate for these variations. Although these mechanisms are poorly understood, it is thought that they elaborate neuronal representations in the inferotemporal cortex that are sensitive to object form but substantially invariant to other image variations. This study examines this hypothesis for image variation resulting from changes in object position. We studied the effect of small differences (+/-1.5 degrees ) in the retinal position of small (0.6 degrees wide) visual forms on both the behavior of monkeys trained to identify those forms and the responses of 146 anterior IT (AIT) neurons collected during that behavior. Behavioral accuracy and speed were largely unaffected by these small changes in position. Consistent with previous studies, many AIT responses were highly selective for the forms. However, AIT responses showed far greater sensitivity to retinal position than predicted from their reported receptive field (RF) sizes. The median AIT neuron showed a approximately 60% response decrease between positions within +/-1.5 degrees of the center of gaze, and 52% of neurons were unresponsive to one or more of these positions. Consistent with previous studies, each neuron's rank order of target preferences was largely unaffected across position changes. Although we have not yet determined the conditions necessary to observe this marked position sensitivity in AIT responses, we rule out effects of spatial-frequency content, eye movements, and failures to include the RF center. To reconcile this observation with previous studies, we hypothesize that either AIT position sensitivity strongly depends on object size or that position sensitivity is sharpened by extensive visual experience at fixed retinal positions or by the presence of flanking distractors. PMID- 12783960 TI - Human frontal eye fields and visual search. AB - Recent physiological recording studies in monkeys have suggested that the frontal eye fields (FEFs) are involved in visual scene analysis even when eye movement commands are not required. We examined this proposed function of the human frontal eye fields during performance of visual search tasks in which difficulty was matched and eye movements were neither necessary nor required. Magnetic stimulation over FEF modulated performance on a conjunction search task and a simple feature search task in which the target was unpredictable from trial to trial, primarily by increasing false alarm responses. Simple feature search with a predictable target was not affected. The results establish that human FEFs are critical to visual selection, regardless of the need to generate a saccade command. PMID- 12783961 TI - Single-pulse transcranial magnetic stimulation of parietal and prefrontal areas in a memory delay arm pointing task. AB - Fifteen healthy volunteers performed a memory-pointing task using their right arm while single-pulse transcranial magnetic stimulation (TMS) above motor threshold was applied over the posterior parietal or prefrontal cortex of the left or right hemisphere in four blocks of trials. The stimulation was randomly delivered at one of three time intervals during the 3-s delay period (early: 300 ms, intermediate: 1,500 ms, late: 2,700 ms). A separate block with no stimulation was used as control. Only early left parietal stimulation resulted in an increase in the variance of movement amplitude but not direction for all targets in two dimensional space (both hemifields). The results point to the significance of the contralateral posterior parietal cortex early on during the memorization of the target for an upcoming movement. Taking into consideration the limitations of TMS and those imposed by the particular task, the lack of specific effects of prefrontal stimulation provides evidence that these areas might not be involved in the performance of simple memorized arm movements. PMID- 12783962 TI - Commissural inputs to secondary vestibular neurons in alert cats after canal plugs. AB - Gaze is stabilized during head movements primarily by the vestibuloocular reflex (VOR). After a unilateral canal plug, the VOR's response is reduced. Recovery of the VOR may be brought about by changes in the efficacy of brain stem synapses or by other mechanisms. We measured the responses of horizontal secondary vestibular neurons (HSNs) to stimulation of the contralateral labyrinth. HSN responses in normal alert cats were compared with those in cats that had recovered from unilateral horizontal semicircular canal (HSCC) plugs. After recovery, excitatory commissural inputs to HSNs on the plugged side elicited significantly smaller responses than in normal cats with no change in mean discharge rates. However, mean discharge rates tended to be higher after recovery for cells receiving inhibitory commissural inputs. The change in resting rate invalidates any direct comparison of inhibitory inputs. These results are interpreted in terms of possible mechanisms for recovery from unilateral vestibular loss by the VOR neural network. We conclude that after unilateral HSCC plugs, changes in brain stem excitatory synapses and/or excitability of secondary vestibular neurons may participate in the restoration of normal vestibular reflexes. PMID- 12783963 TI - Best practice in thyroid pathology. AB - Thyroid pathology is a specialist area but is often encountered by the general pathologist in a variety of forms including cytology, frozen sections, and resection specimens. In the thyroid gland, as for other endocrine organs, many aspects of diagnosis are unique to this area of histopathology; thus, the aims of this paper are to set out best practice guidelines which, although not entirely comprehensive, will be of practical use. PMID- 12783964 TI - Bone marrow immunohistology of plasma cell neoplasms. AB - The application of immunohistology to the spectrum of plasma cell disorders has yet to be incorporated widely into routine haematology practice. This technique enables the direct visualisation of specific surface and cytoplasmic antigens in the context of the individual cell and the surrounding anatomical neighbourhood. This review outlines the role of bone marrow immunohistology in the laboratory evaluation of patients with suspected and established plasma cell neoplasms and its emerging role in understanding myeloma biology for possible future therapeutic application. PMID- 12783965 TI - Myogenin and MyoD1 expression in paediatric rhabdomyosarcomas. AB - The diagnosis of paediatric solid tumours is often based on small tissue needle biopsies in which many different entities demonstrate a "small round cell tumour" phenotype and in which there may be insufficient tissue to allow the interpretation of diagnostic architectural features, which may be present in larger specimens. Therefore, the extensive use of a panel of immunohistochemical markers is part of the routine handling and investigation of such biopsies to reach a definite diagnosis. However, in some cases the morphological and routine immunohistochemical findings may be insufficient for a precise diagnosis or they may be difficult to interpret in the given clinical context. Although many paediatric tumours exhibit characteristic chromosomal translocations with resultant specific fusion transcripts, these require molecular methods for their detection, usually on fresh tissue samples, which may not always be available. As more immunohistochemical markers become available, more precise diagnosis on such small biopsies may be possible. This review examines the use of the immunohistochemical markers, MyoD1 and myogenin, in the diagnosis of paediatric rhabdomyosarcoma, including its subtypes. PMID- 12783967 TI - Immune escape mechanisms in ALCL. PMID- 12783968 TI - The Caldicott report and patient confidentiality. PMID- 12783966 TI - The immunopathogenesis of meningococcal disease. AB - This review describes the mechanisms of the immune response to meningococcal disease, examining the extent to which individual variation of the immune response can determine susceptibility. It concludes by summarising the difficulties encountered by recent efforts to develop new immunomodulatory treatments. PMID- 12783969 TI - Increased incidence of cytomegalovirus but not Chlamydia pneumoniae in atherosclerotic lesions of arteries of lower extremities from patients with diabetes mellitus undergoing amputation. AB - AIMS: To evaluate the association between cytomegalovirus (CMV) or Chlamydia pneumoniae infection and the development of accelerated atherosclerotic lesions in patients with diabetes who are known to have an impaired immune response to infection and a high incidence of atherosclerosis. METHODS: Two hundred arterial samples from patients with diabetes who had undergone surgical amputation for gangrenous lower limbs were selected to assess the presence of CMV or C pneumoniae nucleic acid by means of the polymerase chain reaction. RESULTS: CMV nucleic acid sequences were detected in 64 of 200 (32%) samples and C pneumoniae in seven of 200 (3.5%) arterial samples with severe atherosclerosis. Of those positive for C pneumoniae, six were also positive for CMV. CONCLUSION: The significantly higher incidence of CMV nucleic acid sequences in the arterial samples of patients with diabetes supports the hypothesis that this organism is involved in the pathogenesis of atherosclerosis in patients with diabetic mellitus. It is possible that the potential role of different infectious agents in the pathogenesis of atherosclerosis might rely on their biological properties and their infectivity in hosts with varying immunological status. PMID- 12783970 TI - Digital slide and virtual microscopy based routine and telepathology evaluation of routine gastrointestinal biopsy specimens. AB - AIMS: To evaluate a recently developed digital slide and virtual microscope system, and to compare this method with optical microscopy on routine gastrointestinal biopsy specimens in both local and remote access modes. METHODS: A fully computer controlled commercial microscope was used. The scanning program included object detection, autofocus, and image compression algorithms. The overall hard disk space for a gastric biopsy was between 30 and 50 MB and the scanning time was between 20 and 40 minutes. Haematoxylin and eosin stained routine gastric (61) and colon (42) biopsy specimens were selected, scanned, and evaluated by two specialists on an optical (OM) and virtual microscope (VM). RESULTS: The overall concordance of VM and OM with the consensus diagnosis was 95.1% and 97%, respectively. Clinically important concordance was 96.1% and 98% for VM and OM, respectively. The two methods showed concordance in 92% of cases and clinically important concordance in 94.1% of cases. The reasons for discordance were image quality (one case), interpretation difference (three cases), and insufficient clinical information (three cases). Remote evaluation of the digital slides through the Internet has the advantages of the previously used static and dynamic telepathology methods. CONCLUSIONS: Diagnostic concordance was found between OM and VM. The digital slide and the virtual microscope can be alternative techniques in the computerisation of the histology laboratory and in teleconsultation services after further evaluation of time and storage constraints. PMID- 12783971 TI - Lymphoproliferative disorders in Oxford renal transplant recipients. AB - BACKGROUND: Increased cancer incidence, particularly lymphoproliferative disease, is a complication of immunosuppression in organ transplantation. Non-Hodgkin's lymphomas (NHLs) occur frequently during the first year after transplantation, more so in North America than in Europe. METHODS: This study audited and correlated the demographic, clinical, pathological, and outcome features of post transplant lymphoproliferative disorders (PTLDs) in a large centre in Oxford, and assessed whether the time of onset fitted more with the European or North American pattern. RESULTS: There were 1383 renal transplants in the study period and 27 patients developed lymphoma: 26 NHLs and one Hodgkin's disease (1.95%). Four of the patients never received cyclosporin. The mean time of diagnosis after transplant was 46 months. Most tumours (21/27) presented extranodally. Management included reduction of immunosuppression, surgical excision, antiviral treatment, radiotherapy, and chemotherapy. Three patients presented in the first post transplant year-0.34% of cyclosporin managed patients-similar to the North American incidence, although the incidence of extranodal late PTLDs was also high (mean onset, 36 months v 15 months international mean). Post-transplant lymphomas were the most common malignancy associated with death in transplant patients. CONCLUSIONS: PTLDs occurred in 2% of renal transplant patients, presenting both in the first year in association with cyclosporin use, as in North America, but also in subsequent years, giving an overall presentation time later than the international mean. The disease usually presented extranodally, accounting for the wide range of symptoms and signs. Despite awareness and active management, the disease contributed to death in more that 50% of patients with PTLDs. PMID- 12783972 TI - Proliferation markers and DNA content analysis in urinary bladder TaT1 urothelial cell carcinomas: identification of subgroups with low and high stage progression risks. AB - AIMS: To evaluate whether in situ biomarkers Ki67, mitotic activity index (MAI), p53, mean area of the 10 largest nuclei (MNA10), and whole genome DNA ploidy by flow and image cytometry (FCM and ICM, respectively) have independent prognostic value in urinary bladder urothelial cell carcinomas (UCs). METHODS: Ki67 and p53 immunoquantitation was performed in TaT1 consensus diagnosis UCs. MAI and MNA10 were also determined. Single cell suspensions were stained (DAPI for FCM; Feulgen for ICM). There was enough material for all measurements in 171 cases. Kaplan Meier curves and multivariate survival analysis (Cox) were used to assess the prognostic value of all features (including classic clinicopathological risk factors, such as stage, grade, multicentricity, carcinoma in situ). RESULTS: Thirteen (7.6%) patients progressed. Of the classic factors, grade was strongly prognostic in univariate analysis, as were all the biomarkers. In multivariate analysis, the strongest independent combinations for progression were MNA10 (threshold (T) = 170.0 micro m(2)) plus MAI (T = 30), or MNA10 (T = 170.0 micro m(2)) plus Ki67(T = 25.0%). p53 (T = 35.2%) plus Ki67 (T = 25.0%) also predicted progression well, with high hazard ratios, but p53 measurements were not as reproducible as the other features. The prognostic value of the quantitative biomarkers exceeded that of the classic risk factors and DNA ploidy. The sensitivity, specificity, positive, and negative predictive values of MNA10/MAI or MNA10/Ki67 at the thresholds mentioned were 100%, 79%, 57%, and 100%, respectively. These feature combinations were also strongest prognostically in the high risk treatment subgroup. CONCLUSIONS: The combined biomarkers MNA10/Ki67 or MNA10/MAI are more accurate and reproducible predictors of stage progression in TaT1 UCs than classic prognostic risk factors and DNA ploidy. PMID- 12783975 TI - Mucinous adenocarcinoma with superficial stromal invasion and villous adenoma of urachal remnants: a case report. AB - This report describes a case of mucinous adenocarcinoma with superficial stromal invasion and villous adenoma originating in the dome of the urinary bladder. Although no urachal remnants were identified, the location suggested urachal derivation. Only two previous cases of urachal adenocarcinoma with features of early stromal invasion associated with a villous tumour have been described. PMID- 12783973 TI - Density of neoplastic lymphoid infiltrate, CD8+ T cells, and CD1a+ dendritic cells in mycosis fungoides. AB - BACKGROUND/AIMS: CD8+ T cells and epidermal/dermal dendritic cells expressing CD1a are found among neoplastic CD4+ T cells in mycosis fungoides (MF) lesions. This study analysed the relation of CD8+ tumour infiltrating lymphocytes (TILs), CD1a+ epidermal Langerhan's cells (LCs), and dermal dendritic cells (DDCs) to clinicopathological parameters in 46 MF cases. METHODS: Pretreatment diagnostic biopsy specimens of 46 MF cases were submitted to histological analysis and immunohistochemistry. Four histological grades were defined based on the density of the neoplastic infiltrate: grade 1 (mild superficial perivascular infiltrate), grade 2 (moderate superficial perivascular infiltrate with some tendency to confluence), grade 3 (pronounced superficial band-like infiltrate), and grade 4 (deep nodular infiltrate). Epidermotropism was scored as low, moderate, or high. Numbers of CD8+ T cells and of dermal and epidermal CD1a+ cells were scored as 1 (low), 2 (moderate), and 3 (high). Correlations between these parameters and clinical data (age, sex, clinical type of lesions, stage, response to treatment, and recurrence) were analysed by the chi(2) test. RESULTS: Numbers of TILs and DDCs were associated with subepidermal infiltrates, being lower in less dense infiltrates, whereas there was no association between epidermal CD1a+ cells and the analysed parameters. Complete remission in treated patients was related to subepidermal infiltrates but not to TILs, LCs, or DDCs. CONCLUSIONS: These results support the notion that CD8+ cells and dermal CD1a+ cells are active against tumour cells. MF with low numbers of TILs could represent an early stage of the disease, before TILs are activated against tumour specific antigens. PMID- 12783974 TI - Histopathological characteristics of small diameter melanocytic naevi. AB - BACKGROUND/AIMS: The clinical definition of an atypical naevus ("dysplastic naevus" or "naevus with architectural disorder and cytological atypia of melanocytes") stresses size larger than 5 mm in diameter as a major diagnostic criterion. Because malignant melanomas and their precursors may arise in smaller lesions, a histological study of melanocytic lesions smaller than 4 mm in diameter was conducted to evaluate their histological appearance. METHODS: Two hundred and sixty one naevi smaller than 4 mm in diameter were collected and characterised by histological examination into benign naevi without architectural disorder and naevi with architectural disorder and mild, moderate, and severe atypical melanocytes according to criteria used on larger lesions. RESULTS: Small melanocytic naevi covered the same complex histological spectrum from benign naevi to severely atypical naevi when compared with larger lesions. A high proportion of small naevi (72%) exhibited features diagnostic for naevi with architectural disorder and cytological atypia. CONCLUSION: There is a discrepancy between histological and clinically defined atypical naevi. The same generally accepted criteria for the histological diagnosis of atypical naevi should be used for small melanocytic naevi in addition to large ones. Thus, small naevi exhibiting atypical features on histological examination should be categorised as atypical naevi, regardless of their small diameter. PMID- 12783976 TI - Intravascular lymphomatosis. AB - Intravascular lymphomatosis (IVL) is a rare angiotrophic large cell lymphoma producing vascular occlusion of arterioles, capillaries, and venules. Antigenic phenotyping shows that these lymphomas are mostly of B cell type, and less commonly T cell or Ki-1 lymphomas. The central nervous system and skin are the two most commonly affected organs; patients usually present with progressive encephalopathy with mental status changes and focal neurological deficits and skin petechia, purpura, plaques, and discolouration. Other involved organs include adrenal glands, lungs, heart, spleen, liver, pancreas, genital tract, and kidneys. Bone marrow, blood, cerebrospinal fluid, and lymph nodes are typically spared. Fever of unknown origin is another common presentation. Only one case of IVL presenting with disseminated intravascular coagulation and anasarca (generalised oedema) has been reported in the literature. This report describes a postmortem case of a patient with IVL who initially presented with disseminated intravascular coagulation complicated by intracerebral haemorrhage. PMID- 12783977 TI - Derivative chromosome 9 deletions in chronic myeloid leukaemia: interpretation of atypical D-FISH pattern. AB - BACKGROUND/AIMS: New molecular cytogenetic techniques are increasingly applied as a routine investigative tool in haematological malignancies, both at diagnosis and subsequent monitoring. This report describes the interpretation of atypical signal patterns encountered using BCR-ABL dual colour dual fusion fluorescence in situ hybridisation (D-FISH) translocation probes in chronic myeloid leukaemia (CML). METHODS: Interphase FISH experiments were carried out using BCR-ABL D-FISH probes in 46 patients with CML at diagnosis and during subsequent disease monitoring. Atypical hybridisation signal patterns were characterised by molecular cytogenetic techniques and correlated with conventional karyotyping. RESULTS: Two patients showed atypical interphase D-FISH patterns with one orange, one green, and one fusion (1O1G1F) signal. The presence of BCR-ABL gene fusion was documented by a dual colour single fusion (S-FISH) probe. The submicroscopic deletion of the ABL-BCR fusion gene on the derivative chromosome 9 in these cases was subsequently characterised by metaphase FISH on relocated G banded metaphases. CONCLUSIONS: Atypical interphase D-FISH patterns should not be interpreted in isolation and should be considered in conjunction with other cytogenetic or molecular genetic investigations. PMID- 12783978 TI - HLA-G does not have a pathophysiological role in Graves' disease. AB - AIMS: It has been suggested that the non-classic HLA class I molecule HLA-G plays a role in autoimmune disease by protecting tissues from damage by infiltrating cytotoxic T cells. Such infiltration occurs in the thyroid of patients with Graves' disease (GD) and Hashimoto's thyroiditis (HT) and can eventually result in tissue destruction. The aim of the current study was to analyse thyroid tissue and thyrocytes obtained from individuals with autoimmune thyroid disease for the expression of HLA-G. METHODS: HLA-G expression was analysed in thyroid tissue taken from six patients with GD and one with HT by reverse transcriptase polymerase chain reaction. Thyroid tissue samples isolated from six patients with multinodular goitre (MNG) were used as non-autoimmune controls. HLA-G expression was also examined in cultured thyroid follicular cells (TFCs). RESULTS: The expression of HLA-G was not detected in the thyroid gland of patients with either GD, HT, or MNG. Furthermore, HLA-G expression could not be detected in cultured patient TFCs under basal conditions or after stimulation with the proinflammatory cytokines-interleukin 1alpha, interferon gamma, and tumour necrosis factor alpha. CONCLUSIONS: HLA-G expression does not occur in the thyroid of patients with GD, indicating that HLA-G does not play a pathophysiological role in this autoimmune disorder. Although the expression of HLA-G was not detected in the thyroid sample of the patient with HT, a greater sample size would be required to conclude that HLA-G does not have a part to play in this autoimmune thyroid disease. PMID- 12783979 TI - Well differentiated fetal adenocarcinoma of the lung in a 29 year old woman. AB - This report describes a case of well differentiated fetal adenocarcinoma of the lung in a 29 year old female smoker. The histological pattern and immunohistochemical profile were consistent with well differentiated fetal adenocarcinoma and the patient made an uneventful postoperative recovery with no recurrence after 18 months. This neoplasm is a rare lung tumour that is composed of glycogen rich neoplastic glands and tubules that resembles fetal lung at 10 to 15 weeks of gestation. It is important to identify this rare variant of adenocarcinoma because it is a low grade malignancy with low associated mortality. PMID- 12783980 TI - Optimal detection of Campylobacter spp in stools. PMID- 12783981 TI - Suboperonic regulatory signals. AB - In prokaryotes, the genome is necessarily small in size, thus creating challenges for gene regulation. Adhya discusses how polycistronic operons can be regulated at the suboperonic level to allow genes to be independently regulated within an operon. This permits the cells to respond to different environmental conditions and allows the genes within operons to encode proteins involved in divergent cellular processes and still be regulated according to the cell's needs. Suboperonic control leads to discoordinate gene expression and can occur through transcriptional regulatory events or translational regulatory events mediated by proteins or cis- or trans-acting RNAs. PMID- 12783982 TI - Analysis of the mobility of signaling molecules in lymphocytes using fluorescence photobleaching techniques. AB - The earliest biochemical events at the plasma membrane that lead to gene activation appear to depend not only on the local concentration of signaling molecules, but also on the mobility of these molecules at the site of signaling. To elucidate the process of signal transduction after receptor engagement in the immune system, it is important to analyze the mobility of signaling molecules in living lymphocytes. Current knowledge of the changes in intracellular localization and dynamic movements of signaling molecules during lymphocyte activation is limited. Here, we describe a method for known as fluorescence recovery after photobleaching, used to measure the diffusion mobility of a signaling molecule in a T cell line after T cell receptor stimulation. This method is a useful tool in studies of spatiotemporal regulation in immunoreceptor signaling. PMID- 12783983 TI - Gene expression profile of aging in human muscle. AB - Studies of gene expression related to aging of skeletal muscle have included few subjects or a limited number of genes. We conducted the present study to produce more comprehensive gene expression profiles. RNA was extracted from vastus lateralis biopsies obtained from healthy young (21-27 yr old, n = 8) and older men (67-75 yr old, n = 8) and was analyzed with high-density oligonucleotide arrays. Of the approximately 44,000 probe sets on the arrays, approximately 18,000 yielded adequate signals for statistical analysis. There were approximately 700 probe sets for which t-tests or rank sum tests indicated a difference (P 75 years of age [38 with ageing-associated cognitive decline (AACD)] and 15 age-matched controls. A 1.5-tesla General Electric MRI scanner was used. Standardized visual ratings were undertaken of white matter hyperintensities (WMH). Grey matter volumes were assessed using voxel-based morphometery. RESULTS: Associations were identified between processing speed and executive function and the severity of WMH in key areas. In addition, atrophy in the fronto-subcortical circuits was associated with AACD. CONCLUSION: Attentional and executive impairments are underpinned by WMH in fronto-striato-thalamo frontal circuits. Frontal atrophy is identified as a novel substrate of cognitive decline in stroke patients. PMID- 12784037 TI - 'Friction and adhesive forces'--possible underlying causes for temporomandibular joint internal derangement. AB - Since normal temporomandibular joint (TMJ) movements depend primarily on the disc freely sliding down the slope of the eminence, understanding how aberrations in the lubrication system contribute to TMJ dysfunction is clearly critically important. It provides a possible explanation for the genesis of disc displacement and helps make us familiar with the clinical appearance and ways of treating limited mouth opening caused by the anchored disc phenomenon (ADP) versus disc displacement without reduction, TMJ open lock versus dislocation and osteoarthritis. This understanding clarifies the efficiency of procedures such as joint hydraulic pump, arthrocentesis and arthroscopic lavage and lysis particularly in ADP, open lock and osteoarthritis. PMID- 12784038 TI - Subchondral bone resorption in temporomandibular joint disorders. AB - Several tissues are involved in temporomandibular joint (TMJ) health, including synovial fluid, the TMJ disc, articular cartilage, and subchondral bone. This article focuses upon bone resorption in temporomandibular joint disorders (TMD) and has the following objectives: (1) to provide a brief review of the current understanding of bone formation and bone resorption (bone remodeling); (2) to present selected case studies which illustrate the spectrum of bone resorption patterns in TMD patients of various ages; (3) to review previous reports in the literature describing loss of subchondral bone in TMD, and (4) to discuss the interaction between osteoporosis and TMD and the potential role for antiresorbing agents in TMD therapy. PMID- 12784039 TI - Nitric oxide in experimental joint inflammation. Benefit or detriment? AB - The host response to infection or injury initiates a cascade of events involving recruitment of leukocytes and the release of multiple inflammatory mediators. One of these mediators, nitric oxide (NO), not only represents an important microbicidal agent in host defense, but also functions as a biological signaling and effector molecule in inflammation and immunity. However, overproduction of NO can be autotoxic and contribute to tissue damage and has been implicated in pathogenesis of tumors, and infectious, autoimmune and chronic degenerative diseases. NO is generated via constitutive and inducible nitric oxide synthases (iNOS) which catalyze the oxidation of a guanidino nitrogen associated with L arginine. Whereas endothelial NOS (eNOS) and neuronal NOS (nNOS) are constitutively expressed, iNOS is transcriptionally induced by bacterial constituents and inflammatory mediators, including TNF alpha and IL-1. In an experimental model of bacterial component-induced joint inflammation and tissue degradation, functionally distinct roles of the constitutive NOS and iNOS were demonstrated. Following systemic delivery of an arthritogenic dose of streptococcal cell walls (SCW), these bacterial peptidoglycan-polysaccharide complexes disseminate and target the peripheral joints, liver and spleen of the treated animals. Following deposition of the SCW in the peripheral joints, an initial innate inflammatory response to the bacterial components progresses into an adaptive immune response with the recruitment and activation of mononuclear phagocytes and T lymphocytes. With the release of cytokines and inflammatory mediators, there is an upregulation of gene expression for iNOS, but not the constitutive nNOS or eNOS. Nonetheless, the constitutive NOS isoforms, regulated by calcium fluxes and interaction with calmodulin, may also enhance NO production. Increased release of NO was detected not only in the synovium, but also in the circulation, and plasma levels of nitrate plus nitrite, the stable products of NO reactions, correlated with disease progression. Following inhibition of NO production with nonspecific NOS inhibitors, such as N(G) monomethyl-L-arginine, which target all three isoforms, there is a striking therapeutic benefit with reduced signs and symptoms of erosive arthritis. In contrast, selective targeting of iNOS with N-iminoethyl-L-lysine resulted in exacerbation of the synovial inflammation and degradation of joint structures. Based on these data, it appears that the constitutive isoforms of NOS contribute to the pathophysiology of the arthropathy, and that induced NOS and NO may function, in part, in a protective pathway. Moreover, the suppression of NO following treatment with TNF alpha antagonists results in reduced inflammation and the associated synovial pathology. Collectively, these data implicate discrete roles for the NOS isoforms in the emergence of local tissue pathology and underscore the need to define the specific pathways that are being targeted for interventional strategies. PMID- 12784040 TI - Pathophysiological mechanisms in osteoarthritis lead to novel therapeutic strategies. AB - Osteoarthritis (OA) is a debilitating, progressive disease of diarthrodial joints associated with aging. At the molecular level, OA is characterized by an imbalance between anabolic (i.e. extracellular matrix biosynthesis) and catabolic (i.e. extracellular matrix degradation) pathways in which articular cartilage is the principal site of tissue injury responses. The pathophysiology of OA also involves the synovium in that 'nonclassical' inflammatory synovial processes contribute to OA progression. Chondrocytes are critical to the OA process in that the progression of OA can be judged by the vitality of chondrocytes and their ability to resist apoptosis. Growth factors exemplified by insulin-like growth factor-1, its binding proteins and transforming growth factor-beta contribute to anabolic pathways including compensatory biosynthesis of extracellular matrix proteins. Catabolic pathways are altered by cytokine genes such as interleukin-1 (IL-1) and tumor necrosis factor-alpha (TNF-alpha) which are upregulated in OA. In addition, IL-1 and TNF-alpha downregulate extracellular matrix protein biosynthesis while concomitantly upregulating matrix metalloproteinase (MMP) gene expression. When MMPs are activated, cartilage extracellular matrix degradation ensues apparently because levels of endogenous cartilage MMP inhibitors cannot regulate MMP activity. Therapeutic strategies designed to modulate the imbalance between anabolic and catabolic pathways in OA may include neutralizing cytokine activity or MMP gene expression or inhibiting signaling pathways which result in apoptosis dependent on mature caspase activity or mitogen-activated protein kinase (MAPK) activity. MAPK activity appears critical for regulating chondrocyte and synoviocyte apoptosis and MMP genes. PMID- 12784041 TI - Analgesic effect of elastoviscous hyaluronan solutions and the treatment of arthritic pain. AB - Elastoviscous hyaluronan solutions have an analgesic effect when injected intra articularly in animal and human joints. This was first discovered using animal behavioral models and later confirmed in neurophysiological studies in cat and rat joints. These studies on both normal and experimentally produced arthritis in joints confirmed that only elastoviscous solutions of hyaluronan or certain of its derivatives (hylans) have a desensitizing effect on nociceptive sensory receptors. Recently, this desensitizing effect of elastoviscous hyaluronan solutions was also demonstrated on intact or on isolated patches of oocyte cell membranes. Viscosupplementation, the exchange of pathological synovial fluid in arthritic joints with pure elastoviscous solutions of hyaluronan or hylans, is a widely accepted therapeutic modality used to provide long-lasting analgesia in human knee joints. The clinical studies performed on human and animal temporomandibular joints since the mid-1970s are reviewed. These trials used three distinctly different preparations made from hyaluronan of different average molecular weight, polydispersity and, consequently, different elastoviscous properties. These differences are demonstrated and the consequences on the potential efficacy of the preparations are discussed. PMID- 12784042 TI - Induction of early growth response gene Egr2 by basic calcium phosphate crystals through a calcium-dependent protein kinase C-independent p44/42 mitogen-activated protein kinase pathway. AB - Using the reverse transcriptase-polymerase chain reaction we examined the effect of basic calcium phosphate (BCP) crystals on the induction of the early growth response gene Egr2 transcription and the signal transduction pathway involved. The results showed that BCP crystals induced Egr2 transcription up to 8-fold, peaking at 24 h after treatment. The induction of Egr2 was confirmed by transient transfection assays using an Egr2 promoter/luciferase reporter construct and could be inhibited by the p44/42 mitogen-activated protein kinase (MAPK)-specific inhibitor U0126, or by calcium chelator TMB-8, but not by the SAPK2/p38 MAPK inhibitor SB202190 or by the protein kinase C inhibitor bisindolylmaleimide I (Bis-I). Using the Mercury Pathway Profiling System (Clontech, Palo Alto, Calif., USA) we further showed that induced Egr2 could stimulate the activities of several transcription factors that are associated with cell proliferation, such as c-fos, SRF and c-myc. PMID- 12784043 TI - Specialized cranial muscles: how different are they from limb and abdominal muscles? AB - Mammalian skeletal muscle fibers can be classified into functional types by the heavy chain (MyHC) and light chain (MyLC) isoforms of myosin (the primary motor protein) that they contain. Most human skeletal muscle contains fiber types and myosin isoforms I, IIA and IIX. Some highly specialized muscle fibers in human extraocular and jaw-closing muscles express either novel myosins or unusual combinations of isoforms of unknown functional significance. Extrinsic laryngeal muscles may express the extraocular MyHC isoform for rapid contraction and a tonic MyHC isoform for slow tonic contractions. In jaw-closing muscles, fiber phenotypes and myosin expression have been characterized as highly unusual. The jaw-closing muscles of most carnivores and primates have tissue-specific expression of the type IIM or 'type II masticatory' MyHC. Human jaw-closing muscles, however, do not contain IIM myosin. Rather, they express myosins typical of developing or cardiac muscle in addition to type I, IIA and IIX myosins, and many of their fibers are hybrids, expressing two or more isoforms. Fiber morphology is also unusual in that the type II fibers are mostly of smaller diameter than type I. By combining physiological and biochemical techniques it is possible to determine the maximum velocity of unloaded shortening (V(o)) of an individual skeletal muscle fiber and subsequently determine the type and amount of myosin isoform. When analyzed, some laryngeal fibers shorten at much faster rates than type II fibers from limb and abdominal muscle. Yet some type I fibers in masseter show an opposite trend towards speeds 10-fold slower than type I fibers of limb muscle. These unusual shortening velocities are most probably regulated by MyHC isoforms in laryngeal fibers and by MyLC isoforms in masseter. For the jaw-closing muscles, this finding represents the first case in human muscle of physiological regulation of kinetics by light chains. Together, these results demonstrate that, compared to other skeletal muscles, cranial muscles have a wider repertoire of contractile protein expression and function. Molecular techniques for reverse transcription of mRNA and amplification by polymerase chain reaction have been applied to typing of single fibers isolated from limb muscles, successfully identifying pure type I, IIA and IIX and hybrid type I/IIA and IIA/IIX fibers. This demonstrates the potential for future studies of the regulation of gene expression in jaw-closing and laryngeal muscles, which have such a variety of complex fiber types fitting them for their roles in vivo. PMID- 12784045 TI - Effect of dialysis on plasma total antioxidant capacity and lipid peroxidation products in patients with end-stage renal failure. AB - Dialysis is associated with an impairment of antioxidant defense and an overproduction of oxidative stress markers. This study focuses on the comparison of plasma total antioxidant capacity (TAC) and lipid peroxidation products in patients on hemodialysis (HD) before and after treatment and in peritoneal dialysis (PD) patients. Plasma TAC, malonaldehyde (MDA), and 4-hydroxyalkenal concentrations were measured in 31 HD patients, in 24 PD patients, and in 17 normal controls (NC). It was found that before HD, TAC and MDA levels were higher than those in the NC (p < 0.001). After HD, these levels decreased significantly but were higher than in NC and lower than in PD patients (p < 0.001). The levels of 4-hydroxyalkenals were elevated in patients as compared with NC, but did not differ between HD and PD patients. The MDA concentrations correlated positively with the TAC in the patients. It is concluded that the oxidative status of patients on HD is similar to that of patients on PD and that the susceptibility to oxidative stress is strongly related to the levels of MDA produced in plasma. PMID- 12784044 TI - Sex differences in rabbit masseter muscle function. AB - The proportions of fibers of different phenotypes in the rabbit masseter muscle differ strikingly in adult males and females. Muscles from females contain similar proportions of small fibers that express both the slow/beta and cardiac alpha myosin heavy chain (MyHC) isoforms and larger fibers containing the IIa MyHC isoform. In muscles from males, nearly 80% of fibers are of the IIa phenotype. To evaluate the functional significance of these sex differences, we used finely graded intramuscular microstimulation to study the contractile properties of masseter motor units in >6-month-old male and female rabbits. Twitch forces and torques in males were significantly greater in magnitude than those of females. Greater proportions of units that produced larger forces/torques were encountered in the males. The same motor units produced force or torque more rapidly in males than in females, principally because units in which twitch rise times were >22 ms were found only in females. The forces applied to the mandible and the torques generated about the right mandibular condyle were studied during cortically evoked rhythmic activation of the masticatory muscles. The overall range of torque rise times and the magnitudes of the peak torques did not differ between sexes. The mean rise time was significantly shorter and the mean peak torque was significantly greater in males. We conclude that sex differences in fiber phenotype proportions are reflected in sex differences in motor unit properties and in the function of these motor units during rhythmic activation. PMID- 12784046 TI - Antioxidant and oxidative stress indices in dialysis-dependent acute renal failure. AB - BACKGROUND: Experimental animal models and in vitro studies have established a role for reactive oxygen species and the therapeutic potential for free radical scavengers in acute renal failure (ARF). Little is known of the effects of hemodialysis and other clinical variables on antioxidant defenses and oxidative stress among patients with ARF. METHODS: We examined antioxidant defenses and oxidative stress status in 24 patients with ARF requiring hemodialysis (HD). Blood samples were drawn prior to the first dialysis session (baseline), as well as before and after the third and sixth dialysis sessions. At each time point, the following parameters were measured: plasma alpha-tocopherol (vitamin E), plasma glutathione peroxidase (GSH-Px), serum total oxygen radical absorbance capacity (ORAC), plasma thiobarbituric acid reactive substances (TBARS), plasma tumor necrosis factor-alpha (TNF-alpha), and interleukin-10 (IL-10). Multivariate linear regression analyses were performed to examine clinical/laboratory variables associated with antioxidant/oxidative stress indices. The changes in antioxidant/oxidative stress indices over time after initiation of hemodialysis was evaluated in a subgroup of patients (n = 11) who completed six dialysis sessions. Intradialytic changes in antioxidant/oxidative stress indices and the differential impact of cellulose acetate vs. polysulfone dialyzers were also evaluated. RESULTS: Factors associated with alpha-tocopherol level were serum albumin (301 microg/dl upward arrow for each 1 g/dl upward arrow in albumin) and ORAC (188 microg/dl upward arrow for each 1,000 micromol Trolox Eq/l upward arrow in ORAC). Age was independently associated with plasma GSH-Px levels (55 U/l downward arrow for each 10-year age upward arrow ). Factors associated with ORAC were alpha-tocopherol (85 micromol Trolox Eq/l upward arrow for each 100 microg/dl upward arrow in alpha-tocopherol) and total bilirubin (30 micromol Trolox Eq/l downward arrow for each 1 mg/dl upward arrow in total bilirubin). Total bilirubin was independently associated with TBARS (0.2 microM upward arrow for each 1 mg/dl upward arrow in total bilirubin). GSH-Px and ORAC levels declined over time between baseline and the sixth dialysis session (p < 0.05 for both). Finally, there was a significant intradialytic decline in ORAC levels, which appeared to be more pronounced with use of cellulose acetate compared with polysulfone dialyzer membranes (p < 0.05). CONCLUSIONS: These observations indicate that antioxidant and oxidative stress indices in ARF patients are associated with several clinical and laboratory variables as well as the dialysis procedure. Further studies are needed to investigate the therapeutic role of anti oxidant therapy in these patients. PMID- 12784047 TI - Influence of relative hypoparathyroidism on the responsiveness to recombinant human erythropoietin in hemodialysis patients. AB - BACKGROUND/AIM: It has been shown in a few studies examining small patient groups that high levels of intact parathyroid hormone (iPTH) were associated with a less efficient response to recombinant human erythropoietin (rHuEPO). However, the responsiveness to rHuEPO in hemodialysis (HD) patients with relative hypoparathyroidism remains undetermined. This study examines the responsiveness to rHuEPO in HD patients with relative hypoparathyroidism. METHODS: We retrospectively studied 19 nondiabetic patients (mean age 44.3 +/- 8.2 years, age range 29.4-55.6 years) treated with HD for chronic glomerulonephritis. Of the 19 patients, 8 (group A) had iPTH levels <100 pg/ml for the preceding 6 months without administration of 1,25-(OH)(2)-vitamin D(3). Eleven patients had iPTH levels >100 pg/ml (group B). Hematocrit (Hct) and rHuEPO doses were recorded for statistical analysis. RESULTS: In patients of groups A and B, the rHuEPO dose (U/kg/week) was 55.21 +/- 16.23 vs. 84.08 +/- 24.56 (p = 0.01); Hct (%) 33.29 +/- 1.72 vs. 31.43 +/- 2.98 (p = 0.67), and rHuEPO resistance index (weekly rHuEPO dose/Hct) 81.38 +/- 16.64 vs. 155.63 +/- 42.22 (p < 0.001). Furthermore, weekly rHuEPO dose and rHuEPO resistance index correlated positively with serum iPTH levels (R = 0.765, p < 0.001; R = 0.764, p < 0.001), whereas the Hct correlated negatively with serum iPTH levels (R = -0.400, p = 0.045). The alkaline phosphatase level (IU/l) was lower (50.46 +/- 12.87 vs. 69.61 +/- 20.68, p = 0.17) in group A. CONCLUSION: Our observations suggest that the lower the iPTH levels of chronic HD patients, even with relative hypoparathyroidism, the better the responsiveness to rHuEPO. PMID- 12784048 TI - A switch to high-flux helixone membranes reverses suppressed interferon-gamma production in patients on low-flux dialysis. AB - Long-term hemodialysis (HD) induces an inflammatory response and is associated with a suppressed cellular immune response manifested, in part, by impaired interferon (IFN-gamma) production. We investigated the effect of high-flux HD using the synthetic Helixone membrane and ultrafiltered dialysate on plasma levels of inflammatory mediators and on the whole blood production of IFN-gamma. METHODS: Twelve ESRD patients were dialyzed under low-flux HD (polysulfone F6) and again after 6 weeks of high-flux HD (Helixone FX100). Ultrafiltered bicarbonate dialysate without bacterial growth and no detectable endotoxin was used throughout the study. Plasma levels of urea, albumin, beta(2)-microglobulin (beta(2)-m), interleukin (IL)-6, C-reactive protein (CRP), IL-1 receptor antagonist (IL-1Ra), IL-18, and IL-18-binding protein (IL-18BP) were measured. In addition, the Staphylococcus epidermidis-induced production of IFN-gamma and IL 18 was assessed in whole blood cultures of HD patients as well as in 9 healthy subjects. RESULTS: Plasma levels of urea, albumin, IL-6, IL-1Ra and CRP were not significantly different between high-flux and low-flux HD. In contrast, beta(2)-m levels decreased significantly by 31% with high-flux Helixone (p < 0.002). Stimulated whole blood production of IFN-gamma was reduced in low-flux HD but increased to near normal levels after 6 weeks of high-flux HD. Plasma levels of free IL-18 and its specific inhibitor IL-18BP were not different between the two dialyzer membranes. CONCLUSION: Compared to low-flux polysulfone HD with ultrafiltered dialysate, high-flux HD with the synthetic Helixone membrane did not result in a significant change in plasma levels of proinflammatory (IL-6, CRP, IL-18) and anti-inflammatory (IL-1Ra, IL-18BP) cytokines. However, high-flux HD restores whole blood IFN-gamma production without significant changes in free IL-18. Therefore, the immune modulation in high-flux HD is likely due to removal of inhibitors of IFN-gamma production other than IL-18BP. PMID- 12784049 TI - Liver support--a task for nephrologists? Extracorporeal treatment of a patient with fulminant Wilson crisis. AB - BACKGROUND: Patients with Wilson's disease may present with cirrhosis, acute hepatitis or fulminant hepatic failure. Without urgent orthotopic liver transplantation, a fulminant Wilson crisis has a mortality of 100%. We report on an 18-year-old female patient with fulminant hepatic failure due to Wilson crisis. METHODS: The molecular adsorbent recirculating system (MARS) was used to eliminate albumin-bound toxins and to bridge waiting until an organ became available. RESULTS: A total of 18 MARS sessions and 4 plasma exchange sessions were performed. Bilirubin levels and hepatic encephalopathy improved under MARS therapy. A total of 75 mg copper was removed until serum copper levels were within the normal range. Copper elimination was measured in 15 MARS treatments, which removed a total of 12.9 mg copper. Four plasma exchange sessions, with a total exchange of 11 liters of plasma, removed 12 mg copper. Urinary copper elimination with penicillamine was 50 mg. CONCLUSION: MARS was an effective method to stabilize a patient with Wilson crisis, contributed to copper elimination and gained time for liver transplantation. The risk of high-urgency transplantation could be avoided. Liver support was easy in the hands of nephrologists familiar with extracorporeal therapy. PMID- 12784050 TI - Changes in circulating levels of calcitonin gene-related peptide and nitric oxide metabolites in septic patients during direct hemoperfusion with polymyxin B immobilized fiber. AB - BACKGROUND/AIMS: This study evaluated the mechanism of hemodynamic improvement in polymyxin B-immobilized fiber (PMX) treatment. METHODS: Fifteen septic patients with endotoxemia and/or gram negative infection were treated with PMX. Plasma concentrations of calcitonin gene-related peptide (CGRP) and nitric oxide metabolites were measured. RESULTS: CGRP and nitric oxide metabolites before treatment were significantly higher than in healthy controls. CGRP had a significantly inverse correlation with diastolic blood pressure and a positive correlation with the Septic Severity Score and the number of failed organs. Systolic and diastolic blood pressure and systemic vascular resistance index increased significantly both immediately after and 24 h after PMX treatment. CGRP decreased significantly 24 h after the treatment. In contrast, there were no significant changes in nitric oxide metabolites. CONCLUSION: PMX treatment improved hemodynamic parameters in septic patients, and CGRP increased with increasing severity of sepsis. We conclude that a decrease in CGRP levels may be related to hemodynamic improvement resulting from PMX treatments. PMID- 12784051 TI - Detoxifying capacity and kinetics of the molecular adsorbent recycling system. Contribution of the different inbuilt filters. AB - The molecular adsorbent recycling system (MARS) represents a cell-free, extracorporeal, liver assistance method for the removal of both albumin-bound and water-soluble endogenous toxins. The aim of the present study was to evaluate the short- and long-term removal capacity and selectivity of the different inbuilt dialysers and adsorption columns (uncoated charcoal, anion exchanger resin). Levels of endogenous toxins and parameters of hepatic synthesis and necrosis were therefore monitored before, during and after the MARS treatment phase in 10 patients. Moreover, blood and dialysate clearances of urea, creatinine, bilirubin and bile acids were determined during a single treatment. The significant increasing time course of total bilirubin blood levels before the start of the treatment could be stopped and reversed in a significant decreasing time course. The removal rates of urea nitrogen, bilirubin and bile acids during a single treatment amounted to 55.5 +/- 4.0, 28.3 +/- 3.9, and 55.4 +/- 4.0% (mean +/- SEM), respectively. The efficacy of removal of albumin-bound toxins sharply declined early after initiation of the treatment to become negligible after 6 h. In conclusion, both albumin-bound and water-soluble toxins are adequately removed by the MARS. Our data suggest that the rate and efficacy of removal of albumin bound toxins are related to both the strength of the albumin binding and the saturation of the adsorption columns. PMID- 12784052 TI - Role of hypoxia in the pathogenesis of renal disease. AB - Despite a high overall oxygen supply, the tissue oxygen tensions in the kidney are comparatively low and render the kidneys prone to hypoxic injury. However, the role of hypoxia in the pathogenesis of different types of renal disease remains incompletely understood. The importance of hypoxic cell injury is most obvious in renal vascular disease, in which occlusion of the renal artery or one of its branches can induce tissue necrosis. In acute renal failure, circumstantial evidence suggests that hypoxic injury to the renal medulla plays a significant role. In addition, chronically impaired oxygenation may also be an important factor in the progression of chronic renal disease. Destruction of the glomerular capillaries leads to hypoperfusion of the peritubular interstitium. Moreover, in focal disease, a compensatory increase in perfusion of other glomeruli may increase flow and pressure in peritubular capillaries derived from their vasa efferentia which could be a cause of microvascular injury. The interstitial capillary density is reduced in chronic renal disease, and results of animal experiments suggest that this is due to an imbalance in the expression of pro- and antiangiogenic factors. Besides its essential role in energy generation, oxygen is increasingly recognized as an important regulator of cellular functions. Hypoxia induces specific genes through increased expression of hypoxia-inducible transcription factors (HIF). Different HIF isoforms have recently been shown to be inducible in glomerular, tubular, and interstitial cells of the kidney. While the majority of HIF-dependent genes confer protection against hypoxia, hypoxia-inducible gene expression has been suggested to contribute also to increased interstitial matrix deposition. PMID- 12784053 TI - Interleukin 18 and interleukin 18 binding protein: possible role in immunosuppression of chronic renal failure. AB - Although interleukin (IL)-18 is a member of the IL-1 family of ligands, IL-18 appears to have unique characteristics, particularly in the regulation of the T helper type 1 (Th1) response. Th1 responses are required for tumor surveillance, killing intracellular organisms, and to provide help for antibody production. In patients with chronic renal failure, the well-known immunosuppression contributes to a failure to respond to infectious challenges and vaccinations. The most salient biological property of IL-18, linking this cytokine to the Th1 response, is its ability to induce interferon gamma (IFN-gamma). In fact, IL-18 was originally identified as an IFN-gamma-inducing factor, and IFN-gamma production is the hallmark of the Th1 response. Dysregulation of IFN-gamma production resulting from reduced activity of IL-18 would explain one of the mechanisms of immunosuppression in patients with chronic renal failure. The activity of IL-18 can be regulated by the IL-18-binding protein (IL-18BP), a glycoprotein of 40,000 daltons, which is constitutively expressed and appears to be the natural inhibitor of IL-18 activity. Unlike soluble receptors for IL-18, IL-18BP does not have a transmembrane domain; IL-18BP is a secreted protein possessing a high affinity binding and ability to neutralize IL-18. IL-18BP was discovered in human urine and is excreted in health following glomerular filtration. With decreasing renal function, the concentrations of IL-18BP in the circulation are elevated as compared with subjects with a normal renal function, and these elevated levels may result in a decreased IL-18 activity. Because of the importance of IL-18 and IFN-gamma in the Th1 response, the biology of IL-18 and IL-18BP is reviewed here in the context of the immunosuppression of chronic renal failure. PMID- 12784054 TI - Arginine vasotocin interacts with the social environment to regulate advertisement calling in the gray treefrog (Hyla versicolor). AB - Arginine vasotocin (AVT) has different effects on social behaviors across species with different social systems. However, less attention has been directed towards the way environmental stimuli interact with AVT to affect social behaviors. Previous work found that AVT affects the production of advertisement calls and the outcome of competitive interactions between male gray treefrogs (Hula versicolor). We conducted a field study to investigate how the presence of other males in a chorus influenced the production of advertisement calls in males treated with AVT or saline. After hormone treatment we placed individuals in call sites either within 50 cm of another calling male (proximate), or more than 2 m away from any calling male (distant). Males treated with AVT produced significantly longer calls with more pulses when placed at proximate call sites, but not distant call sites. Nonsignificant trends indicated that these increases in call length coincided with a decrease in call rate. There was no effect of call site distance on advertisement calls when males were treated with saline. These results indicate that the social environment can modulate the action of AVT on an important reproductive behavior, and that AVT does not influence call duration and pulse number unless another male is nearby. Understanding how social stimuli regulate the action of AVT on behavior could provide insight on the diversity of AVT function across species. PMID- 12784055 TI - Structure and function of visual displays produced by male jacky dragons, Amphibolurus muricatud, during social interactions. AB - Many lizards produce visual displays to mediate social behavior. However, most studies have focused upon displays used by iguanid lizards. We investigated and quantified the displays used by jacky dragons, an agamid lizard from southeastern Australia. By establishing male jacky dragons within individual territories and conducting experimental presentations of male and female intruders, we were able to demonstrate that territorial male Amphibolurus muricatus utilize a fixed action pattern display comprising discrete motor components to indicate territoriality and aggression. Displays directed to intruders contain an extra introductory component and modifying postural elements, whereas non-directed displays given in the absence of intruders lack these features and appear to be used to advertise territory ownership. Although the sequence of display components remained the same across non-directed, male-directed and female directed displays, resident males differed in pushup duration and the number of pushups per display in relation to individual body size. This display variance might function to provide an honest signal revealing the body size or the physical condition of each individual, permitting opponents to assess each other effectively through their displays. Visual signals produced by intruding lizards were qualitatively distinct from the displays given by residents, and appear to indicate submission. However, male and female intruders differed in the frequency with which each type of submissive gesture was performed, implying that this might convey intruder sex. PMID- 12784056 TI - Steroid correlates of territorial behavior in male jacky dragons, Amphibolurus muricatus. AB - Male jacky dragons, Amphibolurus muricatus, indicate territoriality to rivals during the mating season through the use of stereotyped motor displays. The relationship between corticosterone (B) and testosterone (T) and its effects on territorial display expression were investigated in captive lizards. Results demonstrated that territorial display production was most effectively predicted by elevated baseline T levels. This parallels the heightened T concentrations exhibited by males in the field during the spring mating period. The effect of social interaction on B and T levels was also examined by presenting territory holding males with a size-matched male intruder. Resident males exhibited clear differences in the level of territorial response evoked by a male intruder, which were correlated with differences in physiological activity. Males that gave no territorial response had moderate B levels that did not change with social interaction, and significantly lower T levels than males that responded to the intruder with territorial displays. Among displaying males, those exhibiting low levels of territorial responses showed no change in B or T. In contrast, high level territorial responders exhibited acute B increases when faced with an intruder, which might assist in supporting sustained metabolic activity and could possibly reflect differences in the perception of social stress. These same males also exhibited a reduction in T levels at such times, probably due to the acute rise in B. Combined, these results suggest that high circulating T is acting in a preparatory manner to increase the likelihood of producing a territorial response upon engaging in a social encounter. However, once this response has been initiated T may not need to remain elevated to allow continued expression of territorial behavior. PMID- 12784057 TI - Effects of running water on lateral line responses to moving objects. AB - We investigated in goldfish, Carassius auratus, and trout, Oncorhynchus mykiss, how running water affects the responses of afferent fibers in the posterior lateral line nerve and of lateral line units in the brainstem medial octavolateralis nucleus to an object that is moved from anterior to posterior or opposite along the side of the fish. In still water, nerve fibers in both species responded to the moving object with alternating periods of increased and decreased firing rate. Most fibers in goldfish but none in trout discharged bursts of spikes in response to the object's wake. Responses of brainstem units were more variable and less distinct than nerve fiber responses. Bursting activity in response to the object's wake was found in only one brainstem unit. In running water, responses of goldfish nerve fibers were weaker than in still water. This effect was independent of object motion direction. Responses of trout fibers were weaker when the object was moved with the flow but were slightly stronger when the object was moved against the flow. In general, running water affected the responses of goldfish nerve fibers more strongly than the responses of trout fibers. Compared to still water, brainstem units in both species responded more weakly when the object was moved with the flow. When the object was moved against the flow, brainstem responses were on average comparable to those in still water. Measurements of changes in pressure and water velocity caused by the moving object indicate that the observed effects can largely be explained by peripheral hydrodynamic effects. However, physiological differences between goldfish and trout units indicate that the lateral line systems in these two species are adapted to different hydrodynamic conditions. PMID- 12784058 TI - Discrimination of functionally referential calls by laboratory-housed rhesus macaques: implications for neuroethological studies. AB - Prior to examining the neural correlates of auditory cognition with ethologically relevant stimuli, it is first necessary to establish that laboratory-housed animals respond to these stimuli with species-typical responses. Here, we report the results of experiments on laboratory-housed rhesus monkeys using both species typical vocalizations and band-pass noise. Paralleling the approach used in field studies of this species, we used a habituation-discrimination paradigm in which auditory stimuli were presented and a monkey's orienting responses to the stimuli were quantified. In parallel with the results obtained in field studies, we found that laboratory-housed rhesus classified species-typical vocalizations according to their putative referent properties as opposed to similarities in their acoustic morphology. In control experiments, monkeys oriented to band-pass noise but did not categorize differences in the spectral composition of the noise stimuli. These findings support the hypothesis that laboratory-housed rhesus classify, in the absence of training, species-typical vocalizations in a manner comparable to rhesus monkeys living under more natural conditions. As such, species-typical vocalizations are an appropriate and necessary class of stimuli in experiments that explore the neural correlates of auditory cognition in rhesus monkeys from a neuroethological perspective. PMID- 12784059 TI - Molecular skin barrier models and some central problems for the understanding of skin barrier structure and function. AB - A better understanding of the structure and function of the human skin barrier is a prerequisite for a more rational design of transdermal drug administration systems. The study of biological structural organisation is, however, confronted with many difficulties, and interpretations of skin barrier-related data should therefore be done with caution. The recently developed 'single gel-phase model' constitutes an attempt to interpret structural and functional skin barrier data from a biophysical standpoint. PMID- 12784060 TI - Bioavailability of clobetasol propionate in different vehicles. AB - Topical clobetasol propionate is widely used for the treatment of psoriasis. One formulation of clobetasol propionate, Skin Cap, was thought by some practitioners and patients to be more effective than other formulations. Differences in corticosteroid bioavailability could account for differences in efficacy. The purpose of this study is to compare the relative bioavailability of clobetasol propionate from Skin Cap to two FDA-approved formulations of clobetasol propionate (Olux foam and Temovate scalp application). The bioavailability was assessed by measuring the percutaneous absorption of clobetasol propionate in vitro on scalp skin, using the human cadaver skin model. There was no significant difference in the percutaneous absorption of clobetasol propionate between Olux foam and Skin Cap (2.09 and 1.93% at 48 h, respectively) or between Olux foam and Temovate scalp application (3.70 and 3.46%, respectively) when applied under unoccluded conditions. Under occluded conditions, there was greater absorption of clobetasol propionate from the Olux foam as compared with Skin Cap (4.94 and 1.57%, respectively; p < 0.05). The Skin Cap vehicle does not provide more bioavailable clobetasol propionate than currently available, FDA-approved products. Other differences, including those in patient compliance, could account for the perceived efficacy of the product. PMID- 12784061 TI - UV/VIS absorbance allows rapid, accurate, and reproducible mass determination of corneocytes removed by tape stripping. AB - The accurate determination of the mass of the horny layer removed by tape stripping is a decisive prerequisite for the application of this technique in penetration studies. A novel method using optical spectroscopy to determine the amount of stratum corneum (SC) is presented. We could show that the absorbance measured in the visible spectral range accurately reflects the mass of the SC fixed on individual tapes. Furthermore, absorbance measurement allows determination of the absolute mass of corneocyte aggregates on the removed tape strips. Topically applied substances do not disturb the spectroscopic measurements in contrast to the conventionally employed weight determination. Identical results were obtained when performing spectroscopic horny layer quantification independently in two separate institutions. Taken together, this new method is rapid, sensitive, reproducible, and accurate. We anticipate a wide application in penetration studies as well as in dermatopharmacokinetics. PMID- 12784062 TI - Evaluation of the pseudo-absorption method to quantify human stratum corneum removed by tape stripping using protein absorption. AB - Tape stripping is a well-known method to study the barrier function of the stratum corneum (SC) and penetration processes of topically applied substances into the horny layer. The quantification of the removed corneocytes for each tape strip is the prerequisite for these studies. The pseudo-absorption of the corneocytes was proposed as a measure for the quantification of the removed corneocyte aggregates. In this study, the pseudo-absorption of the corneocytes in the visible range (430 nm) is compared with the protein absorption in the UV range (278 nm) and an absorption at 652 nm obtained after staining of the SC proteins with Trypan blue. Both the protein absorption and the absorption measured after staining correlate well with the pseudo-absorption measured at 430 nm (R(2) = 0.92 +/- 0.04 and R(2) = 0.95 +/- 0.04, respectively). PMID- 12784063 TI - Effects of oral bexarotene (targretin) on the minimal erythema dose for broadspectrum UVB light. AB - Photo(chemo)therapy and oral retinoid therapy for psoriasis or cutaneous T-cell lymphoma are frequently combined to obtain an enhanced therapeutic effect with lower safety risks. Bexarotene, a new RXR-selective retinoid (rexinoid), has been developed for the treatment of cutaneous T-cell lymphoma and has recently been investigated in the treatment of psoriasis. In the present study the UV modulating properties of bexarotene were evaluated by assessment of the minimal erythema dose (MED) for UVB light. In 11 patients participating in a phase II study of oral bexarotene 0.5 mg/kg/day (7 patients) or 3.0 mg/kg/day (4 patients) for plaque-type psoriasis, MED tests were performed on uninvolved psoriatic skin on the lower back of the subjects before and after 12-week treatment. Clinical scores of erythema and determination of the MED 24 h after irradiation did not show statistically significant changes between the exposed areas before and after bexarotene treatment or between the two doses tested. No photosensitizing reactions were observed. This study demonstrates that a single exposure to UVB irradiation is well tolerated in patients treated with bexarotene 0.5-3.0 mg/kg/day and suggests that it is not necessary to take precautions with respect to short-term effects of sun exposure during bexarotene treatment. Further study of bexarotene photo(chemo)therapy in cutaneous T-cell lymphoma and psoriasis is warranted. PMID- 12784064 TI - Characteristics of skin wrinkling and dermal changes induced by repeated application of squalene monohydroperoxide to hairless mouse skin. AB - We have studied the effect of squalene monohydroperoxides (Sq-OOH), initial products of UV-peroxidated squalene, on the skin of hairless mice. Repeated topical application of 10 mM Sq-OOH to hairless mice for 15 weeks induced definite skin wrinkling. When image analysis was used to compare wrinkle formation induced by ultraviolet B (UVB) irradiation and Sq-OOH treatment, the degree of wrinkling in exposed skin was seen to be similar. However, the characteristics of wrinkles induced by either method differed markedly with regard to direction and distribution. Biochemical analysis revealed a significant decrease in collagen content per unit area and mass in Sq-OOH-treated skin, whereas no changes per unit area and decrease in collagen per unit mass were observed in UVB-irradiated skin. As for glycosaminoglycan (GAG) content per unit area, significant increases were observed in both Sq-OOH-treated skin and UVB irradiated skin. These changes were not induced by organic hydroperoxides such as TERT-butylhydroperoxide or cumene hydroperoxide treatment. Histological observation revealed epidermal hyperplasia and dermal alterations such as collagen degradation and GAG increases in Sq-OOH-treated skin. Histological changes induced by Sq-OOH were not as pronounced as those induced by UVB irradiation. These results clearly suggest that the wrinkling and changes in dermal collagen content induced by Sq-OOH are qualitatively different to those induced by UVB exposure. This may provide a useful model for the study of skin aging, particularly with regard to collagen content. PMID- 12784065 TI - Aminoglycosides suppress tRNA processing in human epidermal keratinocytes in vitro. AB - The ever-growing resistance of pathogens to antibiotics and the lack of potent antibacterial drugs constitute major problems in the treatment of infectious diseases. Thus, the better understanding of the mode of action of antibiotics at the molecular level is of essential importance. Accumulating evidence points towards RNA as being a crucial target of antibacterial and antiviral drugs. Interestingly, aminoglycosides, one of the most important families of antibiotics, apart from their inhibitory effect on ribosome function, reportedly interfere with various RNA molecules and in vitro suppress the proliferation of human keratinocytes. In this study we investigated the effect of the aminoglycosides neomycin B, paromomycin, tobramycin and gentamycin on ribonuclease P activity from normal human epidermal keratinocytes. All aminoglycosides tested revealed a dose-dependent inhibition of tRNA maturation, which was reduced by increasing Mg(2+) ion concentrations, indicating competition of the cationic aminoglycosides with magnesium ions required for catalysis. Our in vitro findings suggest that the inhibitory effects of aminoglycosides on tRNA processing may be implicated in the mechanisms of their antiproliferative action on human epidermal keratinocytes. PMID- 12784067 TI - Attenuation of axon reflexes to compound 48/80 after repeated iontophoresis of compound 48/80 in skin of the human forearm. AB - The aim of this study was to determine whether the iontophoretic administration of the mast cell degranulator compound 48/80 influences axon reflex vasodilatation in the skin of the human forearm. In stage 1, compound 48/80 was administered by iontophoresis to a circular site in the forearm of 9 healthy men and 8 healthy women on four occasions spread over 24 h. Two control sites were also prepared by passing the iontophoretic current through 0.9% saline. Large wheals initially developed at the compound 48/80 site in 8 of the males and in 2 of the females, but wheals were minimal in all subjects by the fourth administration. In stage 2, compound 48/80 iontophoresis provoked substantial flaring at the first control site, whereas saline iontophoresis induced only minor flaring at the second control site, indicating that compound 48/80 induced axon reflex vasodilatation. However, prior treatment with compound 48/80 inhibited flaring to compound 48/80 in subjects who initially developed wheals, consistent with mast cell degranulation. In stage 3, flaring after iontophoresis of histamine was investigated at the site of compound 48/80 pretreatment and at the second control site in 12 subjects. Flaring was impaired only slightly in 6 subjects who initially developed wheals to compound 48/80. The persistence of flaring indicates that repeated administrations of compound 48/80 did not abolish neurogenic inflammation. Transcutaneous iontophoresis of compound 48/80 may be an attractive alternative to intradermal injection in studies that aim at clarifying the function of mast cells in healthy and diseased skin. PMID- 12784066 TI - Platelet-activating factor antagonist WEB 2086 inhibits ultraviolet-B radiation induced dermatitis in the human skin. AB - It has been suggested that platelet-activating factor (PAF) plays a role in the pathomechanisms of various inflammatory diseases. In an experimental animal model we demonstrated earlier that a selective PAF receptor antagonist gel inhibits ultraviolet-B (UVB) light-induced edema in mouse ears. The goal of our present investigation was to determine whether locally applied WEB 2086, a selective PAF receptor antagonist, alters the dermatitis-causing effect of UVB light on human skin. We induced dermatitis in healthy volunteers by irradiating their skin with UVB light in increasing doses. The irradiated area was treated with WEB 2086 gel (3%) or with a placebo. Erythema was measured spectrophotometrically after 24 and 48 h. After both 24 and 48 h, the WEB 2086 gel significantly inhibited the UVB light-induced erythema at each radiation dose in comparison with the placebo. The PAF antagonist gel therefore proved to be effective against UVB-induced dermatitis. Our results may help to understand the relative importance of mediators in UVB-induced dermatitis and might perhaps pave the way to better therapeutic modalities in this condition. PMID- 12784069 TI - Variability in brain death declaration practices in pediatric head trauma patients. AB - The American Academy of Pediatrics' 'Guidelines for the determination of brain death in children' [Pediatrics 1987;80: 298-300] has been cited since its publication as the definitive reference for all cases of pediatric brain death. As these guidelines appear to have been designed for use in patients where the etiology of coma is unclear, they often seem inappropriate in cases of severe head trauma. We questioned whether these guidelines were truly the national standard of practice, particularly in instances of brain death secondary to head trauma. We conducted a survey of pediatric hospitals and pediatric neurosurgeons across the country regarding this matter, and found that their brain death declaration practices varied widely. The majority of hospitals and neurosurgeons in our survey do not follow the guidelines exactly. We feel that these guidelines should be reevaluated and perhaps revised, especially in the setting of severe head trauma. PMID- 12784068 TI - Complications of intrathecal baclofen pumps in children. AB - Intrathecal baclofen is increasingly being used to manage severe spasticity in children. Although substantial tone reduction with this treatment has been documented, complications also occur. In this study, we describe the device- and major non-device-related complications in a group of 100 consecutive children and young adults who received 117 intrathecal baclofen pumps for the management of severe spasticity. Twenty-four patients (24%) experienced a total of 48 complications. The most common complication was disconnection of the catheter at its connection to the pump, occurring in 9% of pumps implanted. This complication occurred more frequently in pumps with catheter access ports (16%) than in those without ports (2%). Catheter dislodgement from the intrathecal space was the next most common complication, occurring in 8% of pumps implanted (13% of pumps with ports, 4% of pumps without ports). To decrease the occurrence of the most common complications of intrathecal pumps, we now typically implant pumps without catheter access ports, and we use 2-piece catheters. Although the lack of an access port may be a disadvantage for troubleshooting, most complications can be detected in pumps without a port. Patient and family education is critical in preventing serious consequences of baclofen withdrawal resulting from catheter related complications. PMID- 12784070 TI - The first description of a device for repeated external ventricular drainage in the treatment of congenital hydrocephalus, invented in 1744 by Claude-Nicolas Le Cat. AB - An 18th century report of a device for repeated extracranial drainage of cerebrospinal fluid in the treatment of congenital hydrocephalus is reviewed. On 15th October 1744, the French surgeon Claude-Nicolas Le Cat (1700-1768) introduced a specially invented canula into the lateral ventricle of a newborn boy with hydrocephalus. The canula was used as a tap and was left in place for 5 days, until the death of the child. This procedure should be seen as the first documented description of a device for repeated ventricular taps in the treatment of hydrocephalus. PMID- 12784071 TI - Intramedullary schistosomiasis. AB - A case study of intramedullary schistosomiasis in a 10-year-old child is reported. The patient presented with a short history of ascending paraparesis with no sensory loss. Sphincter dysfunction was rapid. She had surgical exploration and removal of a conus medullaris mass. Schistosomiasis was confirmed histologically. Combined steroid and praziquantel therapy improved her sphincter function and paraparesis. PMID- 12784072 TI - Frontal and occipital horn width ratio for the evaluation of small and asymmetrical ventricles. AB - The aim of this study was the introduction of a new linear ventricular measurement which has a better correlation with ventricular volume than the commonly used linear measurements, especially in chronically shunted patients with small and/or asymmetrical ventricles. Ventricular volume, brain volume, Evans ratio, Schierman index, cella media index and frontal and occipital horn ratio were measured from CT scans of 57 shunted hydrocephalic patients and 20 normal control children. Four groups of patients were identified with regards to the ventricular size and symmetry (small, large, symmetrical and asymmetrical). A new linear measurement was defined as half of the ratio of the frontal and occipital ventricular width to that of the skull (frontal and occipital horn width ratio; FOHWR) and compared to other linear measurements in different patient groups. FOHWR showed better correlation with ventricular size and ventricular brain ratio than the ventricular linear measurements currently in use, especially in chronically shunted patients with small and/or asymmetrical ventricles. PMID- 12784073 TI - Alternative uses for the subgaleal shunt in pediatric neurosurgery. AB - The subgaleal shunt has been used for the temporary bypass of the normal cerebrospinal fluid (CSF) pathways. We retrospectively reviewed all subgaleal shunts placed at the Children's Hospital, Birmingham, Ala., USA, from 1997 to the present and examined all uses (e.g. indication, length of follow-up) of the subgaleal shunt outside its use for temporary CSF diversion in premature infants with intraventricular hemorrhage and subsequent hydrocephalus. The average length of survival of the primary subgaleal shunt in this population was 32.2 days. We have had good success with subgaleal shunts in children with malignant brain tumors, intraventricular abscesses, chronic truncal wounds, chronic subdural hygromas and meningitis. However, the greatest utility has been in those scenarios in which the peritoneal cavities were not currently, but with time would be, candidates for distal shunt implantation. Examples of these instances are patients with hydrocephalus and necrotizing enterocolitis or hydrocephalus and preoperative abdominal wall pathology such as omphalocele. PMID- 12784074 TI - Nonclosure of the peritoneum at shunt insertion. AB - To evaluate the nonclosure of the peritoneum during insertion of the distal end of a ventriculoperitoneal shunt in the peritoneal cavity, 43 patients in whom a ventriculoperitoneal shunt was inserted using laparotomy with nonclosure of the peritoneum for insertion of the distal catheter were reviewed. No morbidity occurred during a follow-up period of 3-9 months. We think that nonclosure of the peritoneum is safe in shunt surgery, but needs to be evaluated in further studies. PMID- 12784075 TI - Hemorrhagic complications of intracranial pressure monitors in children. AB - Intracranial pressure (ICP) monitoring plays a valuable role in the management of head injuries and other causes of raised ICP in the pediatric population. The purpose of this study was to investigate the incidence of hemorrhage after ICP monitor insertion, and to classify these complications in a clinically relevant manner. Hospital charts of 431 children (ages 0-16 years) admitted to a level I trauma center over a 2-year period were reviewed and 112 patients (134 insertions) who underwent intraparenchymal ICP monitoring were identified. The authors reviewed postoperative neuroradiological studies. One hundred and nineteen procedures were carried out without any hemorrhage (grade 0). After 10 insertions, a small punctate hemorrhage or localized subarachnoid hemorrhage occurred (grade 1). Three patients sustained an intracerebral hemorrhage that did not require evacuation or manifest as a new neurological deficit (grade 2). There were no hemorrhagic complications that necessitated evacuation or resulted in a noticeable change in the patient's clinical condition (grade 3). We propose a new grading system for hemorrhage after ICP monitor insertion. We found a complication rate close to 10% in our pediatric patients. Fortunately, these hemorrhages were clinically silent and no neurosurgical intervention was necessary. However, grade 1 and grade 2 hemorrhages may manifest with a false reading of high ICP, and the long-term consequences of these complications are not known. Of note, only 23% of these complications were reflected in the patients charts, which may explain the low complication rates reported in other studies that did not analyze postoperative neuroradiological studies. PMID- 12784076 TI - Recurrence of pediatric cerebral arteriovenous malformations after angiographically documented resection. AB - Angiographically confirmed surgical resection is believed to be the 'gold standard' for cure in the treatment of intracranial arteriovenous malformations (AVMs). However, rare exceptions of recurrent AVMs have been documented. We are aware of 11 reported cases of recurrent AVMs in which complete resection of the initial lesion was confirmed by postoperative angiography. Eight of these cases were in the pediatric population. In this report, we present 2 additional cases of such recurrent lesions, review the clinical and scientific literature on this rare phenomenon and provide suggested management guidelines. PMID- 12784077 TI - Application of diffusion tensor imaging to magnetic-resonance-guided brain tumor resection. AB - Interventional magnetic resonance imaging (MRI) continues to make a profound impact on the practice of neurosurgery. We describe a new MRI modality, diffusion tensor imaging (DTI), which uses the diffusion energy of water to map white matter fibers. DTI has been established in other disorders such as metabolic, demyelinating and ischemic diseases. We describe the use of DTI in identifying white matter tracts such as optic radiations and avoiding them intraoperatively in 2 children with low-grade glial tumors. PMID- 12784078 TI - Epilepsy surgery in infancy. A review of four cases. AB - It has been demonstrated that intractable seizures in children can be eliminated or become more responsive to anticonvulsant medication after surgical resection of the epileptogenic focus or dysfunctional hemisphere. We describe our surgical experience with 4 infants treated at Children's Healthcare of Atlanta Egleston Hospital between 1994 and 2002. All infants, ranging in age from 5 to 9 months, presented with severe seizure disorders and failed trials of anticonvulsants. All had preoperative EEG monitoring and MRI studies; 2 had PET functional imaging. One infant underwent a temporal resection for a low-grade glioma. The rest had cortical resections for malformations and dysplasia. All had improvement, with 2 infants free of seizures off medication. We argue for early intervention in severe cases, as the potential for recovery can be dramatic. PMID- 12784079 TI - Cortical ependymoma. A case report and review. AB - The authors report a rare case of a cortical ependymoma in a 10-year-old boy. The patient presented with complex partial seizures and a well-circumscribed, right frontal cortical mass. Routine microscopy showed a glial tumor with diverse histologic features. Immunohistochemistry and electron microscopy were required to establish the definitive diagnosis of cortical ependymoma. Cortical-based pediatric brain tumors range from World Health Organization grade I to III lesions and require significantly different treatment and follow-up. This case illustrates the importance of establishing an accurate neuropathologic tissue diagnosis of all pediatric cortical tumors. PMID- 12784080 TI - Unilateral lambdoidal synostosis with mendosal suture involvement. PMID- 12784081 TI - 'Cortical thumb sign' in X-linked hydrocephalus. PMID- 12784082 TI - Alteration of alpha(1A)-adrenoceptor gene expression in the prostate of streptozotocin-induced diabetic rats. AB - Diabetes-associated alterations in prostate alpha(1A)-adrenoceptor (alpha(1A)-AR) gene expression were studied using a streptozotocin (STZ) induced diabetic rat model. Male Wistar rats were divided into four groups: group I animals were vehicle-treated normal rats; group II consisted of vehicle-treated, STZ-diabetic rats; group III represented insulin-treated, STZ-diabetic rats (0.5 IU/kg three times daily for 4 days), and group IV animals were phlorizin-treated, STZ diabetic rats (1 mg/kg three times daily for 4 days). The expression of mRNA that encoded protein of alpha(1A)-AR in the rat prostate was studied using reverse transcription combined with polymerase chain reaction. The alpha(1A)-AR protein expression in the prostate was studied by Western blotting analysis with a polyclonal antiserum. A 2.51 +/- 0.21-fold increase in the mRNA level of alpha(1A)-AR was observed in the prostate of diabetic rats (n = 8, p < 0.05). Similarly, there was a 2.23 +/- 0.10-fold increase in the alpha(1A)-AR protein level (n = 8, p < 0.05). Both insulin and phlorizin treatments restored the normal levels of mRNA and protein expression. In conclusion, the gene expression of alpha(1A)-AR is increased in the prostate of diabetic rats. Hyperglycemia plays a major role in this alteration. PMID- 12784083 TI - Preservation of endothelial function by the HMG-CoA reductase inhibitor fluvastatin through its lipid-lowering independent antioxidant properties in atherosclerotic rabbits. AB - Recent evidence suggests that the beneficial effects of 3-hydroxy-3 methylglutaryl coenzyme A reductase inhibitors on entothelial function and cardiovascular ischemic events may be attributed not only to their lipid-lowering effects but also to cholesterol-lowering independent (direct) effects on the atherosclerotic vessel wall. This study was designed to test the hypothesis that fluvastatin (Flu) preserves the endothelial function by its cholesterol-lowering independent actions. Rabbits were fed a 0.5% high-cholesterol (HC) diet for 12 weeks (progression phase) and then fed the HC diet either containing or not containing Flu 2 mg/kg/day for an additional 8 weeks (treatment phase). Rabbits fed a normal diet were used as controls. Plasma total and low-density lipoprotein cholesterol concentrations did not differ during the treatment phase: Endothelium dependent/NO-mediated relaxation (acetylcholine and A23187) was impaired in the HC diet group, whereas it was preserved in the HC plus Flu treatment group. The endothelium-independent relaxation (sodium nitroprusside) was similar between the three groups. Interestingly, aortic oxidative stress (lipid peroxides and isoprostane F(2alpha)-III contents) and NADPH oxidase component (p22phox and gp91phox) mRNA expression were increased in the HC group but not in the HC plus Flu group. The A23187-induced nitric oxide production from the aorta was increased in both HC and HC plus Flu groups. There was no significant difference in tissue endothelial-type nitric oxide synthase mRNA expression. Plaque area and intimal thickening of the aorta were significantly lowered in the HC plus Flu group. Flu treatment preserved the endothelial function associated with the decrease in markers of oxidative stress in this experiment. These beneficial endothelial effects of Flu are likely to occur independently of plasma lipid concentrations and to be mediated by its antioxidant action. PMID- 12784084 TI - Mechanisms underlying the contractile response to endothelin-1 in the rat renal artery. AB - We assessed the functional response and the mechanisms following receptor stimulation of endothelin-1 (ET-1) in the rat renal artery. In this study, isometric tension was recorded in renal artery rings without endothelium. Cumulative application of ET-1 from 0.1 to 100 nmol/l induced a sustained concentration-dependent contraction in the renal artery. Submaximal contraction induced by 10 nmol/l ET-1 in 2.5 mmol/l Ca(2+) and in the absence of inhibitors was used as control response (100%). The relative contribution of different sources of Ca(2+) in ET-1-induced contraction was evaluated. The contractile response to 10 nmol/l ET-1 in 2.5 mmol/l Ca(2+ )(1.2 +/- 0.2 g) was significantly inhibited either in Ca(2+)-free solution containing 100 micromol/l ethylene glycol bis-(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (0.6 +/- 0.1 g) or after depletion of intracellular Ca(2+) stores (0.62 +/- 0.05 g). The contribution of phospholipase C and protein kinase C was evaluated by using their inhibitors 2-nitro-4-carboxyphenyl N,N-diphenylcarbamate (NCDC) and [1-(5 isoquinolinesulfonyl)-2-methylpiperazine] (H-7), respectively. The contractile response to 10 nmol/l ET-1 was inhibited by 10 micromol/l NCDC (to 80 +/- 6%) and 30 micromol/l H-7 (to 76.6 +/- 6.5%). We found that 1 micromol/l nifedipine inhibited the ET-1-induced contraction (to 48.7 +/- 6.9%), indicating the contribution of Ca(2+) influx through voltage-gated L-type Ca(2+) channels to this response. Further, the inhibitory effect of nifedipine was to a greater extent as compared with NCDC or H-7. Additive inhibition of ET-1-induced contraction was not observed in the presence of both nifedipine and NCDC. We also evaluated the role of the ionic transport system in the ET-1-induced response by using 20 nmol/l 5-(N-ethyl-N-isopropyl)-amiloride (EIPA), an inhibitor of Na(+) H(+) exchange, or 100 micromol/l ouabain, an inhibitor of Na(+)-K(+)-ATPase. The response to ET-1 was decreased by both EIPA (to 61.6 +/- 8.4%) and ouabain (to 62.1 +/- 8.6%). The contribution of Na(+)-Ca(2+) exchange to ouabain action was tested using the inhibitor dimethyl amiloride HCl (10 micromol/l). The decrease in ET-1-induced contraction by the combination of ouabain and dimethyl amiloride HCl was similar to that observed with ouabain alone. In view of these observations, both extra- and intracellular sources of Ca(2+) contribute to the contractile response induced by ET-1 in the renal artery. Our findings also revealed the importance of Ca(2+) influx through voltage-gated L-type Ca(2+) channels in mediating contraction to ET-1 in the renal artery, whereas a minor role of phospholipase C and protein kinase C was observed. Na(+)-H(+) exchange and Na(+)-K(+)-ATPase also play a role in the ET-1-induced contraction in renal artery. Moreover, the contribution of Na(+)-K(+)-ATPase in ET-1 contraction is not an Na(+)-Ca(2+) exchange-related process. PMID- 12784085 TI - Comparison between CL-316243- and CGP-12177A-induced relaxations in isolated canine ureter. AB - We compared the effects of CL-316243, a selective beta(3)-adrenoceptor agonist, and CGP-12177A, a nonconventional partial beta-adrenoceptor agonist, on the KCl induced contraction in the isolated canine ureter. CL-316243 concentration dependently relaxed the ureteral contraction, the pD(2) value being 7.75 +/- 0.11. This relaxation was competitively antagonized by the selective beta(3) adrenoceptor antagonist SR58894A and by the nonselective beta-adrenoceptor antagonist bupranolol, their pA(2) values being 7.08 +/- 0.08 and 6.43 +/- 0.09, respectively. CGP-12177A concentration dependently reduced the KCl-induced contraction, the pD(2) value being 6.30 +/- 0.25. Even at 1 x 10(-5) mol/l, CGP 20712A (a selective beta(1)- adrenoceptor antagonist) did not shift the concentration-response curves for CL-316243 or CGP-12177A. SR58894A did not induce a parallel rightward shift in the concentration-response curve for CGP 12177A, but bupranolol did produce such a shift, pA(2) and slope values in the Schild plot being 7.15 +/- 0.77 and 0.60 +/- 0.15, respectively. Hence, the competition characteristics for SR58894A and bupranolol differed between the CL- 316243-induced and CGP-12177A-induced relaxations. Our results suggest that CGP 12177A produces ureteral relaxation in the dog via an atypical beta-adrenoceptor (possibly, an atypical site/state of the beta(3)-adrenoceptor) as well as via the typical beta(3)-adrenoceptor. PMID- 12784086 TI - Effects of nonsteroidal anti-inflammatory drugs on prostacyclin and thromboxane in the kidney. AB - Dose-response curves were obtained relating the effects of increasing amounts of aspirin, a nonselective cyclooxygenase (COX) inhibitor, and celecoxib, a selective cyclooxygenase 2 (COX-2) inhibitor, on the concentrations of prostacyclin and thromboxane in renal cortex and medulla of rabbits. The concentrations of the two agonists (aspirin and celecoxib) which elicit a half maximal response on the prostanoid concentration (EC(50)) were compared. Additionally, controls for prostacyclin and thromboxane were related to values for the experimental groups. The EC(50) values for celecoxib were considerably lower than those for aspirin, indicating that celecoxib was more effective in suppressing prostanoid production. There were also significant differences between the majority of experimental groups and their respective controls, further evidence for the greater inhibitory activity of celecoxib on prostacyclin. Celecoxib lowered the ratio prostacyclin/thromboxane in the renal medulla; mercuric chloride further diminished the concentration of prostacyclin in the renal medulla. The results confirm that in the normal rabbit kidney, both nonselective and specific COX inhibitors interfere with renal prostanoid synthesis, but that a selective COX-2 inhibitor is more effective. PMID- 12784087 TI - Imipramine-induced antinociception in the formalin test. Receptor mechanisms involved and effect of swim stress. AB - This study concerned the effect of swim stress on imipramine-induced antinociception in mice. The data showed that intraperitoneal (i.p.) administration of different doses of imipramine (10-40 mg/kg) and 0.5-3 min of swim stress (17 degrees C) induced antinociception in the first and second phases of the formalin test. Low period of swim stress (10 s) with low doses of imipramine (2.5, 5 and 10 mg/kg i.p.), which did not have any effect by themselves, in combination showed antinociception in the second phase of the test. Either yohimbine (0.5 mg/kg i.p.) or naloxone (1 mg/kg i.p.) reversed the response induced by the combination of low doses of imipramine plus swim stress. Yohimbine (1 mg/kg i.p.) decreased the response of imipramine (20 mg/kg i.p.) but not that of 30 s swim stress in the second phase. However, naloxone (1 mg/kg i.p.) reduced the antinociception induced by imipramine (20 mg/kg i.p.) or 30 s swim stress in the second phase of the test, the combination of imipramine with swim stress was not altered by yohimbine or naloxone. Prazosin induced antinociception by itself in the first phase of the test and increased swim stress-induced antinociception with no interaction. It is concluded that antinociception induced by imipramine in the second phase of formalin test may be mediated through alpha(2)-adrenoceptor antagonists. The results indicate that the responses of swim stress and imipramine may be mediated by an opioid mechanism, but the combination of both drugs induced higher antinociceptive effects. PMID- 12784088 TI - Effects of magnesium sulfate on tissue lactate and malondialdehyde levels after cerebral ischemia. AB - In the present study, the effects of magnesium sulfate on tissue lactate and malondialdehyde (MDA) levels after cerebral ischemia in rabbits were studied. The rabbits were divided equally into three groups. Group 1 (n = 8) was the sham operated control group, in group 2 (n = 8) only cerebral ischemia was induced by clamping bilaterally the common carotid arteries for 60 min, and in group 3 (n = 8) magnesium sulfate was administered at a dose of 100 mg/kg i.v. within 5 min after opening the clamps. In group 1 EEG recordings were obtained immediately and 60 and 120 min after craniectomy. In groups 2 and 3 EEG recordings were obtained immediately after craniectomy but before clamping and 60 min after clamping. One hour after opening the clamps and taking EEG recordings, brain cortices were resected, and the concentrations of lactate and MDA were determined using spectrophotometric/enzymatic and thiobarbituric acid methods, respectively. In all groups, there were significant differences between MDA and lactate levels (p < 0.05). There were no significant differences in lactate levels between groups 2 and 3 (p > 0.05), and also the preischemic EEG grades were the same in all groups. Preischemic and postischemic EEG values were significantly different (p < 0.05), and there were also significant differences between postischemic EEG grades in groups 2 and 3 (p < 0.05). There was a correlation between postischemic EEG grades and MDA and lactate levels. These results demonstrate that cerebral ischemia-reperfusion injury leads to an increase in brain tissue lactate and MDA levels, that magnesium sulfate suppresses the increase of MDA and lactate concentrations, and that magnesium sulfate treatment improves the EEG changes. The EEG grades correlated well with MDA and lactate levels. PMID- 12784089 TI - Effects of nasal calcitonin on bone mineral density following parathyroidectomy in patients with primary hyperparathyroidism. AB - AIM: To investigate whether nasal salmon calcitonin (CT; 200 U/day) given in addition to calcium helps to restore the bone mass after parathyroidectomy (PTX) in patients with primary hyperparathyroidism (PHPT). METHODS: Twenty patients with PHPT were enrolled after successful PTX and received 1 g calcium per os daily for 1 year. They were randomly assigned either to nasal CT (CT group) or to no treatment. The bone mass was measured using dual-energy X-ray absorptiometry at multiple sites. RESULTS: Eight patients in each group completed the study. After 12 months, the bone mass increased significantly at whole-body level and at lumbar spine in both groups, increased at hip and epiphyses of tibia or radius in the CT group only, and did not change at diaphyses of tibia and radius in either group. CONCLUSIONS: Bone mass increases after PTX for PHPT in patients receiving oral calcium. CT may help to restore the bone mass at sites of the appendicular skeleton, where trabecular bone predominates. PMID- 12784090 TI - Androgen therapy in constitutional delay of growth. AB - BACKGROUND/AIMS: Two modalities of androgen therapy prevail in the treatment of constitutional delay of growth (CDG): monthly injections of testosterone or daily tablets of the non-aromatizable oxandrolone. The present study was undertaken to prospectively compare both compounds and dose. METHODS: Thirty patients with CDG were the subjects of this study. The protocol required that they all be at age 12 14 years with a bone age delay of more than 2 'years', height less than -2 SDS and growth velocity less than -0.5 SDS. The subjects were at a Tanner stage 1 or 2 and testicular volume were no larger than 4 ml. They were randomly assigned into 3 treatment groups: group 1 patients received monthly injections of 25 mg testosterone propionate-enanthate; group 2 patients received monthly injections of 50 mg testosterone propionate-enanthate; group 3 patients received oral oxandrolone at a weekly dose of 0.7 mg/kg. Treatment was given for a period of 6 months and follow-up commenced 6 months later and yearly thereafter for 2 years. RESULTS: Height velocity and height increased significantly only in groups 2 and 3. Bone age advanced most in group 2. Puberty progressed faster in that group as compared with group 3. The predicted adult height before and 2 years after completion of treatment remained unchanged in the two testosterone groups. It increased significantly in the oxandrolone group from a mean 169.8 cm before therapy to a mean 177.5 cm 2 years after completion of therapy. Peak GH levels were significantly higher on both testosterone 50 mg and oxandrolone, as compared to pretreatment levels. The increment was significantly greater in group 2 as was the increment in serum IGF-1 and IGFBP3. CONCLUSIONS: These results imply that 6 months of testosterone injections at a dose of 50 mg, but not 25 mg, is an effective and safe treatment for patients with CDG, with no considerable impact on final height prediction. On the other hand, daily oxandrolone treatment, starting at age 12-14 years, may increase the predicted final adult height. PMID- 12784091 TI - Insulin-like growth factor-1 receptor regulation in activated human T lymphocytes. AB - OBJECTIVE: To investigate the kinetics of insulin-like growth factor-1 receptor (IGF-1R) expression in PHA-stimulated T lymphocytes. METHODS: IGF-1R protein and mRNA were detected by flow cytometry and RT-PCR respectively, between 0 and 48 h after cell activation. RESULTS: Few minutes after T lymphocytes were activated, internalization of the IGF-1R from the cell membrane was observed, achieving the lower level between 1 and 6 h and was accompanied by a reduction in its mRNA. This was followed by re-expression of IGF-1R on the cell surface and an increase in IGF-1R mRNA levels in the cytoplasm, reaching levels higher than those recorded initially after 48 h activation. CONCLUSION: This down- and up regulation suggests that restoration of IGF-1R would be the result of receptor recycling and de novo synthesis and highlights its importance for T lymphocyte proliferation. PMID- 12784092 TI - A new mutation of 5-alpha-reductase type 2 (A62E) in a large Egyptian kindred. AB - OBJECTIVE: To describe the clinical, biological and molecular data in a large Egyptian kindred with 5alpha-reductase deficiency. PATIENTS AND METHODS: Three patients with ambiguous genitalia were referred at the ages of 20, 9 and 2 years, respectively. In all cases, parents were first cousins. Basal and post-HCG stimulation plasma levels of testosterone and dihydrotestosterone were determined. Direct sequencing and restriction site analysis were applied for patient and family study. RESULTS: A homozygous alanine to glutamic acid substitution at position 62 (A62E) was found in the three patients. The parents and two XX sisters were heterozygous while a third XX sibling was normal. CONCLUSION: We report a new mutation of the 5alpha-reductase type 2 gene. The presence of this mutation in all studied patients and their parents suggests its causative role in 5alpha-reductase deficiency. Identification of the mutation enabled genetic counselling for three XX individuals. PMID- 12784093 TI - Testosterone is significantly reduced in endurance athletes without impact on bone mineral density. AB - AIMS: To compare the basal plasma reproductive hormonal profile in three groups of athletes involved in different training programs, and to define the relationship between androgen level and bone mineral density (BMD) in male athletes. METHODS: Basal serum total testosterone (TT), free androgen index (FAI), sex hormone-binding globulin (SHBG), cortisol, cortisol to TT ratio, luteinizing hormone (LH), estrogen and BMD were evaluated in cyclists (CY; n = 11), triathletes (TR; n = 14) and swimmers (SW; n = 13) and compared with less active controls (n = 10). RESULTS: TT and FAI levels were lower (p < 0.05) in CY and TR, whereas the ratio of cortisol to TT was increased in CY only (p < 0.05). No alteration in serum LH, SHBG, estrogen or cortisol concentration was observed. BMD was higher in the proximal femur in TR (p < 0.05). No BMD or hormonal differences were found in SW. CONCLUSION: Only the endurance training of CY and TR induced androgen deficiency without apparent alteration of BMD. PMID- 12784094 TI - The serum cortisol:cortisone ratio in the postoperative acute-phase response. AB - OBJECTIVE: In a previous cross-sectional pilot investigation, an increase in the ratio of active cortisol to inactive cortisone in serum has been found as a general phenomenon during the acute-phase response. The aim of the present study was to further characterize this alteration of cortisol metabolism in patients undergoing elective cardiac bypass surgery. METHODS: Cortisol and cortisone were quantified by use of liquid-chromatography tandem mass spectrometry in sera that were sampled preoperatively and on the first 4 postoperative days (POD) from 16 patients undergoing aortocoronary bypass grafting (7.00 a.m.). RESULTS: The median serum cortisol concentration peaked on the first POD and then decreased statistically significantly until the end of the observation period: preoperatively, 245 nmol/l (IQR 198-331); 1st POD, 532 nmol/l (IQR 409-678 ); 4th POD, 373 nmol/l (IQR 306-493); p for trend = 0.019. In contrast, the cortisol:cortisone ratio was constantly increased approximately twofold on all POD compared to preoperative sampling: preoperatively, 5.4 (IQR 5.0-7.2); 1st POD, 11.3 (IQR 9.2-13.6); 4th POD, 9.9 (IQR 7.7-11.0), with no significant trend of normalization. CONCLUSION: Following major surgery, the substantial increase in the serum cortisol:cortisone ratio - reflecting a shift in the overall set point of 11beta-hydroxysteroid dehydrogenase activity - is more sustained than the increase in serum cortisol; the increase in the cortisol:cortisone ratio seems to be a long-term phenomenon of the activation of the hypothalamic pituitary-adrenocortical system by surgical stress and systemic inflammation. PMID- 12784095 TI - Successful treatment of partial nephrogenic diabetes insipidus with thiazide and desmopressin. AB - OBJECTIVE: To clarify whether combination treatment with desmopressin (DDAVP) and thiazide was clinically effective in a patient with congenital nephrogenic diabetes insipidus (CNDI), we evaluated the treatment in a 7-year-old boy with CNDI who had demonstrated a partial response to DDAVP. METHOD: Both volume of urine and the presence of nocturia were determined during treatment. RESULT: Neither the usual therapy of a low-salt diet and a thiazide nor intranasal therapy with a large dose of DDAVP was effective. However, combination treatment resulted in a decrease in urinary volume and the disappearance of nocturia. CONCLUSION: DDAVP coupled with thiazide may be useful for CNDI in patients who have shown a partial response to DDAVP. PMID- 12784096 TI - Manifestation of thyrotoxic periodic paralysis in two patients with adrenal adenomas and hyperandrogenaemia. AB - Hypokalaemic periodic paralysis is a fairly common complication of hyperthyroidism in Asian populations, but a rare event in Caucasians. In the present work we describe 2 male Caucasian patients with thyrotoxic periodic paralysis (TPP) as initial clinical manifestation of Graves' disease. Further diagnostic procedures demonstrated unilateral adrenal adenoma and hyperandrogenaemia in both patients. To date, only few data are available concerning the hormonal status of Caucasian patients with TPP. The constellation of TPP and adrenal adenomas with increased levels of androgens has not been described previously. Since androgens are capable of inducing sodium-potassium ATPase, which is thought to be centrally involved in the pathogenesis of TPP, hyperandrogenaemia may have triggered the manifestation of paralytic attacks in our patients. It may be of interest to focus not only on thyroid dysbalances in patients with TTP but also to investigate other hormonal disturbances. PMID- 12784097 TI - Short-term adolescent nicotine exposure has immediate and persistent effects on cholinergic systems: critical periods, patterns of exposure, dose thresholds. AB - In adolescents, the symptoms of nicotine dependence can appear well before the onset of habitual smoking. We investigated short-term nicotine exposure in adolescent rats for corresponding cholinergic alterations. Beginning on postnatal day 30, rats were given a 1-week regimen of nicotine infusions or twice-daily injections, at doses (0.6, 2, and 6 mg/kg/day) set to achieve plasma levels found in occasional to regular smokers. In the cerebral cortex, midbrain, and hippocampus, we assessed nicotinic cholinergic receptor (nAChR) binding, choline acetyltransferase (ChAT) activity, a constitutive marker for cholinergic nerve terminals, and [(3)H]hemicholinium-3 (HC-3) binding to the high-affinity choline transporter, which responds to cholinergic synaptic stimulation. nAChR upregulation was observed with either administration route, even at the lowest dose; in the hippocampus, increases could be detected with as little as 2 days' treatment at 0.6 mg/kg/day. In the midbrain, upregulation was still significant even 1 month post-treatment. Adolescent nicotine treatment also produced lasting decrements in HC-3 binding that were separable from effects on ChAT, suggesting cholinergic synaptic impairment. Again, these effects were obtained at the lowest dose and remained significant 1 month post-treatment. Our results indicate that in adolescence, even a brief period of continuous or intermittent nicotine exposure, elicits lasting alterations in cholinergic systems in brain regions associated with nicotine dependence. As the effects are detected at exposures that produce plasma concentrations as little as one-tenth of those in regular smokers, the exquisite sensitivity of the adolescent brain to nicotine may contribute to the onset of nicotine dependence even in occasional smokers. PMID- 12784098 TI - Effect of CYP2D6 genotypes on the metabolism of haloperidol in a Japanese psychiatric population. AB - We investigated the effect of CYP2D6 genotypes on plasma levels of haloperidol (HAL) and reduced haloperidol (RHAL) in 88 Japanese schizophrenic inpatients being treated with HAL. Some subjects carrying CYP2D6*5 allele (CYP2D6*1/CYP2D6*5, CYP2D6*5/CYP2D6*10) showed extremely high concentrations of both HAL and RHAL, and the groups with CYP2D6*5 allele seemed to have higher plasma concentrations of HAL (1.14+/-0.69 ng/ml/mg) and RHAL (1.10+/-1.05 ng/ml/mg) than the other groups. Among those without CYP2D6*5 allele, there were no significant differences in plasma concentrations of HAL and RHAL between those without CYP2D6*10 allele (HAL=0.68+/-0.31 ng/ml/mg, RHAL=0.28+/-0.37 ng/ml/mg), those with one CYP2D6*10 (HAL=0.70+/-0.23 ng/ml/mg, RHAL=0.31+/-0.16 ng/ml/mg) and those with two CYP2D6*10 alleles (HAL=0.69+/-0.14 ng/ml/mg, RHAL=0.40+/-0.09 ng/ml/mg), although there was a tendency of higher plasma concentration of RHAL in those with two CYP2D6*10 alleles. At a lower daily dosage of HAL (<10 mg/day), the subjects with two or one CYP2D6*10 allele(s) showed significantly higher plasma concentrations of RHAL (0.43+/-0.23 ng/ml/mg, 0.34+/-0.16 ng/ml/mg) than those without CYP2D6*10 allele (0.18+/-0.16 ng/ml/mg). The results of this study indicate that CYP2D6*10 allele plays significant but modest role in HAL metabolism in Japanese; nevertheless, we should not lump CYP2D6*10 allele with CYP2D6*5 allele because these two mutated alleles seem to have different impacts in the metabolism of HAL. PMID- 12784099 TI - Synapse-associated protein 90/postsynaptic density-95-associated protein (SAPAP) is expressed differentially in phencyclidine-treated rats and is increased in the nucleus accumbens of patients with schizophrenia. AB - Phencyclidine (PCP) induces a psychotomimetic state that closely resembles schizophrenia. Therefore, PCP-treated animals can provide a model for schizophrenia. Using differential display, we identified a gene regulated by the delayed action of PCP in rat nucleus accumbens (NAcs). Sequence analysis showed that the cDNA clone obtained was identical to rat synapse-associated protein 90/postsynaptic density-95-associated protein 1 (SAPAP1). Quantitative reverse transcriptase (RT)-PCR analysis showed that SAPAP1 mRNA had increased significantly in rat NAc (P<0.0001) and hippocampus (P<0.01) 24 h after a PCP (10 mg/kg) injection as compared to the controls. Immunoquantification using an anti SAPAP1 antibody indicated that immunoreactivity for SAPAP1 increased significantly (P&<0.05) in the NAcs of unmedicated patients with schizophrenia, as compared to the control subjects and medicated patients with schizophrenia. Our findings support the hypothesis that there is abnormal glutamatergic neurotransmission in schizophrenia and show evidence of abnormalities in the intracellular signal transduction via N-methyl-D-aspartate (NMDA) receptors. PMID- 12784100 TI - Duloxetine increases serotonin and norepinephrine availability in healthy subjects: a double-blind, controlled study. AB - Evidence suggests that compounds that increase the synaptic availability of more than one neurotransmitter have greater efficacy in the treatment of depression than single-acting drugs. Preclinical studies indicate that duloxetine acts to inhibit serotonin (5-HT) and norepinephrine (NE) transporters. The ability of duloxetine to alter 5-HT and NE reuptake was tested in 12 healthy male subjects. Placebo, desipramine 50 mg b.i.d., and duloxetine (80 mg q.d. or 60 mg b.i.d.) were compared in a randomized, double-blind, three-period crossover study in 12 healthy male subjects. Whole-blood 5-HT, urinary excretion of NE and major metabolites, and TYR PD30 (IV tyramine pressor dose needed to increase systolic blood pressure by 30 mmHg) were measured at steady state. Vital signs were measured periodically. Duloxetine affected 5-HT reuptake, with whole-blood 5-HT depletion vs placebo (80 mg q.d.: p=0.07; 60 mg b.i.d.: p=0.02; combined regimens: p=0.01). Cardiovascular changes reflecting increased sympathetic tone were observed with both duloxetine and desipramine, and both treatments significantly decreased whole body NE turnover (p<0.01). Duloxetine and desipramine were associated with similar mean increases in fractional extraneuronal NE concentration, although these changes did not reach statistical significance. TYR PD30 increased significantly with desipramine dosing (p<0.01). In conclusion, whole-blood measurements confirm that duloxetine inhibits platelet 5-HT uptake in vivo. Urinary and cardiovascular measurements suggest that duloxetine has an effect on NE synthesis and turnover, indicative of NE reuptake inhibition. The lack of a detectable impact of duloxetine on TYR PD30 suggests that this may not be the most sensitive indirect measure of NE reuptake when assessing dual reuptake inhibitors. PMID- 12784101 TI - Phencyclidine exacerbates attentional deficits in a neurodevelopmental rat model of schizophrenia. AB - Schizophrenia is characterized by severe abnormalities in cognition, including disordered attention. In the rat, neonatal ventral hippocampal (NVH) lesions induce behavioral abnormalities at adulthood thought to simulate some aspects of the symptomatology of schizophrenia. Here, we compared the effects of NVH and adult ventral hippocampal (AVH) lesions on attentional performance as assessed by the five-choice serial reaction time task (5-CSRTT). NVH-lesioned rats were slower to acquire the task than AVH-lesioned and control animals. When training was complete, NVH- and AVH-lesioned animals exhibited stable but disrupted performance under standard conditions, thus emphasizing an implication of VH in visual attentional processes. Variations in task parameters induced a significantly greater disruption in NVH- and AVH-lesioned groups as compared to controls. NVH-lesioned rats were also hyper-responsive to the disruptive effects of a high dose of phencyclidine (PCP) (3 mg/kg). In contrast, amphetamine (0.4 0.8 mg/kg) had a similar effect in control and VH-lesioned rats. Thus, NVH lesioned rats were impaired in the acquisition of stable performance in the 5 CSRTT, and were hypersensitive to the cognitive-impairing effects of PCP. PMID- 12784102 TI - Stressor controllability modulates stress-induced dopamine and serotonin efflux and morphine-induced serotonin efflux in the medial prefrontal cortex. AB - It has previously been shown that inescapable (IS) but not escapable (ES) stress potentiates the rewarding properties of morphine as measured by conditioned place preference and psychomotor activation, and that this potentiation may be mediated by dorsal raphe nucleus (DRN) serotonin (5-HT) neurons. The medial prefrontal cortex (mPFC) has been implicated in both reward and stress, and is a projection region of the DRN. The mPFC also contains dopaminergic afferents from the ventral tegmental area, which has been the focus of many studies exploring both the rewarding properties of drugs and the aversive properties of stress. The role of the mPFC in stress/drug reactivity interactions is largely unknown. The present study used in vivo microdialysis to examine 5-HT and dopamine (DA) efflux in the mPFC of rats during IS, ES or no stress (NS). IS and ES rats received the stressor in yoked pairs. The stressor consisted of tailshocks that could be terminated for both rats by the ES rats. Large increases in 5-HT and DA levels were observed during IS but not ES or NS. DA and 5-HT efflux were also measured 24 h later in the same rats in response to morphine (3 mg/kg) or saline. Sustained increases in 5-HT levels were observed after morphine in rats that had previously received IS but not in rats that had received ES or NS. No changes in DA efflux were observed after morphine. Thus, 5-HT and DA in the mPFC may be involved in stressor controllability effects, and the sensitization of 5-HT neurons by IS extends to the mPFC and to morphine as a challenge. PMID- 12784103 TI - Opiate withdrawal induces Narp in the extended amygdala. AB - The negative affective states associated with drug withdrawal produce long lasting behavioral effects thought to play a central role in the development and maintenance of dependence. However, little is known about the molecular mechanisms mediating the long-term effects of drug withdrawal. Neuronal activity regulated pentraxin (Narp) is a secreted neuronal immediate early gene (IEG) product that regulates AMPA receptor clustering at synapses. As both IEGs and changes in AMPA receptor trafficking mediate enduring forms of neuronal plasticity, we have assessed whether Narp could be involved in the molecular adaptations accompanying drug withdrawal. To this end, we checked the effect of opiate withdrawal on Narp expression in the extended amygdala, a brain region closely linked to the aversive effects of drug withdrawal. We found a marked increase in the number of Narp-positive cells in this region following opiate withdrawal triggered by either low doses of opiate antagonists or by 'natural withdrawal', removal of the morphine pellets used to induce dependence. In contrast, Arc, another 'effector' IEG, was not induced by opiate withdrawal. As expected, pretreatment of animals with clonidine, which blocks opiate withdrawal, suppresses Narp induction in this paradigm. These results implicate Narp in mediating the long-term, aversive behavioral effects induced by opiate withdrawal. PMID- 12784104 TI - Investigation of epistasis between the serotonin transporter and norepinephrine transporter genes in anorexia nervosa. AB - Weight-restored patients with anorexia nervosa (AN) respond favorably to the selective serotonin reuptake inhibitor fluoxetine, which justifies association studies of the serotonin transporter gene (SLC6A4, alias SERT) and AN. Case control studies suggest that the least transcriptionally active allele of the SERT gene promoter polymorphism (5-HTTLPR) has an increased frequency in AN patients. However, this finding was not replicated with 55 trios (AN child+parents) and the transmission disequilibrium test (TDT). To clarify the role of the 5-HTTLPR in susceptibility to AN, we used the TDT and 106 Australian trios to provide 93% power to detect a genotypic relative risk (GRR) of 2.0. Our results were negative for this GRR (McNemar's chi(2)=0.01, df=1, p=0.921, odds ratio 1.0, 95% CI 0.7-1.5). Additionally, we found no association with AN females, AN subtype, age at onset, or minimum BMI. We then performed the first reported investigation of epistasis between the SERT gene and norepinephrine transporter gene (SLC6A2, alias NET) in AN, as an earlier study suggested that atypical AN responds to the dual serotonin-norepinephrine reuptake inhibitor venlafaxine. We observed no epistasis between the 5-HTTLPR and a polymorphism within the NET gene promoter polymorphic region (NETpPR) (chi(2)=0.48, df=1, p=0.490). Although 5-HTTLPR modulates serotonin reuptake by the serotonin transporter, our analyses provide no evidence that susceptibility to AN is modified by 5-HTTLPR alone, nor in concert with as yet undetermined functional effects of the NETpPR polymorphism. PMID- 12784105 TI - Aripiprazole, a novel atypical antipsychotic drug with a unique and robust pharmacology. AB - Atypical antipsychotic drugs have revolutionized the treatment of schizophrenia and related disorders. The current clinically approved atypical antipsychotic drugs are characterized by having relatively low affinities for D(2)-dopamine receptors and relatively high affinities for 5-HT(2A) serotonin receptors (5-HT, 5-hydroxytryptamine (serotonin)). Aripiprazole (OPC-14597) is a novel atypical antipsychotic drug that is reported to be a high-affinity D(2)-dopamine receptor partial agonist. We now provide a comprehensive pharmacological profile of aripiprazole at a large number of cloned G protein-coupled receptors, transporters, and ion channels. These data reveal a number of interesting and potentially important molecular targets for which aripiprazole has affinity. Aripiprazole has highest affinity for h5-HT(2B)-, hD(2L)-, and hD(3)-dopamine receptors, but also has significant affinity (5-30 nM) for several other 5-HT receptors (5-HT(1A), 5-HT(2A), 5-HT(7)), as well as alpha(1A)-adrenergic and hH(1)-histamine receptors. Aripiprazole has less affinity (30-200 nM) for other G protein-coupled receptors, including the 5-HT(1D), 5-HT(2C), alpha(1B)-, alpha(2A)-, alpha(2B)-, alpha(2C)-, beta(1)-, and beta(2)-adrenergic, and H(3) histamine receptors. Functionally, aripiprazole is an inverse agonist at 5-HT(2B) receptors and displays partial agonist actions at 5-HT(2A), 5-HT(2C), D(3), and D(4) receptors. Interestingly, we also discovered that the functional actions of aripiprazole at cloned human D(2)-dopamine receptors are cell-type selective, and that a range of actions (eg agonism, partial agonism, antagonism) at cloned D(2) dopamine receptors are possible depending upon the cell type and function examined. This mixture of functional actions at D(2)-dopamine receptors is consistent with the hypothesis proposed by Lawler et al (1999) that aripiprazole has "functionally selective" actions. Taken together, our results support the hypothesis that the unique actions of aripiprazole in humans are likely a combination of "functionally selective" activation of D(2) (and possibly D(3)) dopamine receptors, coupled with important interactions with selected other biogenic amine receptors--particularly 5-HT receptor subtypes (5-HT(1A), 5 HT(2A)). PMID- 12784106 TI - Psychoactive drugs and pilot performance: a comparison of nicotine, donepezil, and alcohol effects. AB - The cholinergic system plays a major role in cognitive abilities that are essential to piloting an aircraft: attention, learning, and memory. In previous studies, drugs that enhance the cholinergic system through different pharmacologic mechanisms have shown beneficial effects on cognition; but dissimilar cognitive measures were used and samples were not comparable. A comparison within the same cognitive tasks, within comparable samples appears desirable. Toward this aim, we compared effect sizes (ES) of performance enhancing doses of nicotine (a nicotinic receptor agonist) and donepezil (an acetylcholinesterase inhibitor) as found in our prior work on pilot performance. We also compared cholinergic ES to those of performance-impairing doses of alcohol. In three randomized, placebo-controlled trials, we assessed the flight performance of aircraft pilots in a Frasca 141 simulator, testing I: the acute effects of nicotine gum 2 mg; II: the effects of administration of 5 mg donepezil/day for 30 days; and III: the acute and 8 h-carryover effects of alcohol after a target peak BAC of 0.10%. We calculated the ES of nicotine, donepezil, and alcohol on a flight summary score and on four flight component scores. Compared to placebo, nicotine and donepezil significantly improved, while alcohol significantly impaired overall flight performance: ES (nicotine)=0.80; ES (donepezil)=1.02; ES (alcohol acute)=-3.66; ES (alcohol 8 h)=-0.82. Both cholinergic drugs showed the largest effects on flight tasks requiring sustained visual attention. Although the two tested cholinergic drugs have different pharmacologic mechanisms, their effects on flight performance were similar in kind and size. The beneficial effects of the cholinergic drugs on overall flight performance were large and the absolute (ie nondirectional) sizes were about one fourth of the absolute ES of acute alcohol intoxication and roughly the same as the absolute 8 h-carryover ES of alcohol. PMID- 12784107 TI - Altered glucocorticoid rhythm attenuates the ability of a chronic SSRI to elevate forebrain 5-HT: implications for the treatment of depression. AB - Both glucocorticoids and selective serotonin reuptake inhibitors (SSRIs) alter aspects of 5-HT function including somatodendritic 5-HT1A autoreceptor sensitivity. Many depressed patients prescribed SSRIs have pre-existing flattened diurnal gluococorticoid rhythm. In these patients, interactions between flattened glucocorticoid rhythm and chronic SSRIs, which impact on the SSRI's ability to elevate forebrain 5-HT, may alter clinical efficacy. To address this issue rats underwent implantation of slow-release corticosterone (75 mg pellet s.c.) (to flatten the glucocorticoid rhythm) or sham surgery, and injection of fluoxetine (10 mg/kg/day i.p., 12 days) or vehicle. Using microdialysis in the frontal cortex we found that (21 h after the last injection) extracellular 5-HT was elevated in fluoxetine- or corticosterone-treated animals, but not in those treated with corticosterone plus fluoxetine. In fluoxetine-treated animals, blockade of terminal reuptake by local perfusion of fluoxetine increased 5-HT to the same level as it did in controls, suggesting normal terminal 5-HT release after chronic fluoxetine. However, 5-HT levels following local reuptake blockade in both the corticosterone and corticosterone plus fluoxetine groups were lower than controls, suggesting a corticosterone-induced decrease in terminal release. Finally in fluoxetine, corticosterone, and corticosterone plus fluoxetine groups, there was marked 5-HT1A receptor desensitization, evidenced by attenuation of the decrease in 5-HT release following systemic fluoxetine injection. The data indicate that, despite desensitization of 5-HT1A autoreceptors, concurrent flattened glucocorticoid rhythm compromises the ability of SSRIs to elevate forebrain 5-HT. These findings suggest a potential mechanism for the reduced antidepressant efficacy of SSRIs in those patients with pre-existing glucocorticoid abnormalities. PMID- 12784108 TI - Acute influences of estrogen and testosterone on divergent and convergent thinking in postmenopausal women. AB - Previous studies indicated an enhanced capability of divergent creative thinking in young women during the ovulatory phase, which expressed itself also by an increased dimensional complexity of ongoing electroencephalographic (EEG) activity. Considering the enhanced plasma levels of estrogen and testosterone characterizing the ovulatory phase, we tested whether short-term administration of estrogen or testosterone in postmenopausal women with constantly low levels of gonadal steroids induces similar changes in divergent thinking. In two placebo controlled cross-over studies, healthy postmenopausal women (n=12, in each study, mean age 58 years, range 47-65 years) were treated transdermally over 3 days with estrogen and testosterone, respectively, at doses inducing plasma hormone concentrations comparable with those observed in young women around ovulation. Capabilities of divergent thought and convergent analytical thought, performance on motor perseveration, and verbal memory were examined. EEG activity was recorded while subjects performed on tasks of thinking and during mental relaxation. Estrogen impaired divergent thinking (p <0.01) and enhanced convergent thinking, motor perseveration, and memory for the initial word list (p <0.05 for all tests). In parallel, EEG dimensional complexity was reduced (p <0.05). Overall, these changes indicate an estrogen-induced shift from a "divergent" towards a more "convergent" mode of processing. However, overall less consistent, effects of testosterone were opposite to those of estrogen. It increased performance on some of the divergent thinking tasks (p <0.05), and tended to increase EEG dimensional complexity during divergent thinking. Data indicate a differential sensitivity of modes of thinking to short-term treatment with estrogen and testosterone in postmenopausal women. PMID- 12784110 TI - Antidepressants and emotional processing. PMID- 12784111 TI - The antidepressant clomipramine regulates cortisol intracellular concentrations and glucocorticoid receptor expression in fibroblasts and rat primary neurones. AB - Incubation of LMCAT fibroblasts cells with antidepressants potentiates glucocorticoid receptor (GR)-mediated gene transcription in the presence of cortisol, but not of corticosterone. We have suggested that antidepressants do so by inhibiting the LMCAT cells membrane steroid transporter and thus by increasing cortisol intracellular concentrations. We now confirm and extend this model to primary neuronal cultures. Clomipramine, a tricyclic antidepressant, increased the intracellular accumulation of 3H-cortisol, but not 3H-corticosterone, in LMCAT cells (+80%) and primary rat neurones (+20%). The latter finding is the first demonstration that a membrane steroid transporter is present in neurones. Moreover, verapamil, a membrane steroid transporter inhibitor, reduced the effects of clomipramine on the intracellular accumulation of 3H-cortisol in LMCAT cells. Finally, clomipramine also decreased GR expression (whole-cell Western blot) in LMCAT cells (50% reduction) and primary rat neurones (80% reduction). This GR downregulation can explain the reduced GR-mediated gene transcription previously described under experimental conditions that do not elicit the effects on the LMCAT cells steroid transporter. This work further supports the hypothesis that membrane steroid transporters regulating the access of glucocorticoids to the brain in vivo are a fundamental target for antidepressant action. PMID- 12784112 TI - Null mutation of the arginine-vasopressin gene in rats slows attentional engagement and facilitates response accuracy in a lateralized reaction time task. AB - The neurophysin vasopressin is thought to play an important role in emotional behavior and aspects of cognition in the rat, and the pathophysiology of this system has been implicated in two neurodevelopmental disorders, namely autism and schizophrenia. Genetic deficiency of vasopressin in rats, resulting from a null mutation of the vasopressin gene, causes alterations of brain development with resulting behavioral and neurochemical phenotypes in adulthood. We previously demonstrated that partial vasopressin deficiency (rats heterozygous for the null mutation) produces enhanced visuospatial attention and motor speeding. Here, the results of studies of homozygous Brattleboro rats that are fully vasopressin deficient are reported. We trained subjects to perform a lateralized reaction time task that measures visuospatial divided attention; in task conditions in which the duration of target stimuli was varied from trial to trial, homozygous Brattleboro rats showed a performance phenotype that consisted of more accurate responding for longer duration, and less accurate responding for briefer duration, target stimuli. No differences in response times were measured. Further experiments revealed that two separate processes produced this complex phenotype: a relatively slowed period of attentional engagement (resulting in compromised detection of fast onset-fast offset stimuli) that only partially masks a generally more accurate pattern of responding. These results, taken with earlier data, indicate that vasopressin plays a critical role in regulating visual attention and cognition, either directly, or via early alterations in neurodevelopment. PMID- 12784113 TI - Toluene-induced locomotor activity is blocked by 6-hydroxydopamine lesions of the nucleus accumbens and the mGluR2/3 agonist LY379268. AB - The abuse of volatile inhalants remains a prominent, yet poorly understood, form of substance abuse among youth. Nevertheless, the identification of a mechanism underlying the reinforcing properties of inhalants has been hampered by the lack of a clearly identifiable neural substrate upon which these chemicals act. One ingredient that is common to many abused inhalants is toluene, an organic solvent that is self-administered by nonhuman primates and rodents. Most drugs of abuse have been found to elicit forward locomotion in rats, an effect owing to the activation of mesoaccumbal dopamine (DA) pathways. Thus, the present study was undertaken using two different approaches to determine whether toluene-induced locomotor hyperactivity is also ultimately dependent upon DA neurotransmission in the mesolimbic nucleus accumbens (NAC). Here we report on the effects of 6 hydroxydopamine (6-OHDA) lesions of the NAC or pretreatment with the metabotropic mGlu2/3 receptor agonist LY379268 on toluene-induced locomotor activity. Both procedures, which are known to alter neurotransmission within the NAC, significantly attenuated toluene's locomotor stimulatory effects. These results provide strong support for a central mechanism of action of inhalants, which in the past has been more typically attributed to general nonspecific mechanisms throughout the brain. Moreover, as with other drugs of abuse, the NAC may be the final common pathway subserving toluene's abuse liability. PMID- 12784114 TI - Reduced behavioral effects of cocaine in heterozygous brain-derived neurotrophic factor (BDNF) knockout mice. AB - Brain-derived neurotrophic factor (BDNF) affects the development of brain neurotransmitter systems, including dopamine and serotonin systems that are important for cocaine's rewarding and locomotor stimulatory properties. Human genomic markers within or near the BDNF locus have been linked to or associated with substance abuse. Post-mortem human brain specimens reveal individual differences in the levels of BDNF mRNA and in mRNA splicing patterns. To assess the effects of lifelong alterations in the levels of BDNF expression on a measure of psychostimulant reward, we have compared locomotor stimulant and rewarding effects of cocaine in heterozygous BDNF knockout mice with effects in their wild type littermates. Heterozygous BDNF knockout mice displayed less locomotion during habituation and less locomotion after cocaine injections. Cocaine conditioned place preferences were reduced in the BDNF heterozygotes. These mice displayed no significant difference from saline control values at a dose of 10 mg/kg s.c. cocaine, although they exhibited cocaine-induced preference at a 20 mg/kg dose. These data confirm important roles for BDNF in psychostimulant actions, presumably via neurotrophic effects on dopamine and serotonin systems. Furthermore, these data support suggestions that differences in human BDNF expression may underlie associations between markers near the human BDNF gene locus and drug addiction. PMID- 12784115 TI - Elevated concentrations of CRF in the locus coeruleus of depressed subjects. AB - Research evidence that corticotropin-releasing factor (CRF) plays a role in the pathophysiology of major depressive disorder (MDD) has accumulated over the past 20 years. The elevation of lumbar cerebrospinal fluid (CSF) concentrations of CRF decreased responsiveness of pituitary CRF receptors to challenge with synthetic CRF, and increased levels of serum cortisol in MDD subjects support the hypothesis that CRF is chronically hypersecreted in at least the endocrine circuits of the hypothalamic-pituitary-adrenal (HPA) axis and may also involve other CRF brain circuits mediating emotional responses and/or arousal. One such circuit includes the excitatory CRF input to the locus coeruleus (LC), the major source of norepinephrine in the brain. Furthermore, there are now reports of decreased levels of CRF in lumbar CSF from MDD patients after symptom relief from chronic treatment with antidepressant drugs or electroconvulsive therapy. Whether this normalization reflects therapeutic effects on both endocrine- and limbic associated CRF circuits has not yet been effectively addressed. In this brief report, we describe increased concentrations of CRF-like immunoreactivity in micropunches of post-mortem LC from subjects with MDD symptoms as established by retrospective psychiatric diagnosis compared to nondepressed subjects matched for age and sex. PMID- 12784116 TI - Maintenance efficacy of divalproex in the prevention of bipolar depression. AB - Breakthrough depression is a common problem in the treatment of bipolar disorder. Only one, recently published, double-blind, placebo-controlled trial has examined the efficacy of divalproex in the prevention of depressive episodes in bipolar patients. This report describes, in further detail, the findings from that trial of the effect of divalproex on multiple dimensions of depressive morbidity in bipolar disorder. A randomized, double-blind, parallel-group, multicenter study was conducted over a 52-week maintenance period. Bipolar I patients, who may have been treated with open-label lithium or divalproex and who met recovery criteria within 3 months of onset of an index manic episode, were randomized to maintenance treatment with divalproex, lithium, or placebo in a 2 : 1 : 1 ratio. Adjunctive paroxetine or sertraline for breakthrough depression was allowed in maintenance phase. Outcome measures were the rate of early discontinuation for depression, time to depressive relapse, proportion of patients with depressive relapse, mean change in Depressive Syndrome Scale score, proportion of patients receiving antidepressants, and time in the study. Among patients taking an antidepressant, a higher percentage of patients on placebo than divalproex discontinued early for depression. Patients who were previously hospitalized for affective episodes or took divalproex in the open period relapsed later on divalproex than on lithium during the maintenance period. Divalproex-treated patients had less worsening of depressive symptoms than lithium-treated patients during maintenance. Indices of severity of prestudy illness course predicted worse outcome in all treatment groups. Divalproex improved several dimensions of depressive morbidity and reduced the probability of depressive relapse in bipolar disorder, particularly in patients who had responded to divalproex when manic, and among patients with a more severe course of illness. PMID- 12784117 TI - Chronic lithium treatment inhibits pilocarpine-induced mossy fiber sprouting in rat hippocampus. AB - Lithium remains the gold standard in the treatment of bipolar disorder. Long-term treatment with lithium may lead to specific adaptational changes in gene expression that contribute to a neuroprotective effect. In this study, the pilocarpine model of spontaneous limbic epilepsy was used to induce mossy fiber sprouting (axonal growth of the dentate granule cells that synapse on the pyramidal cells of the CA3 region) to examine the prophylactic neuroprotective effects of lithium in vivo. There were four groups of animals: pilocarpine treated (Pil+/Li-); pilocarpine treated followed by lithium (Pil+/Li+); lithium alone (Pil-/Li+); control (Pil-/Li-). Timm staining was used to obtain density measurements in the stratum oriens and the inner molecular layer of the hippocampus. Mossy fiber density was higher in the pilocarpine-treated animals compared to controls. Chronic lithium following pilocarpine treatment attenuated the density of mossy fibers but lithium alone had no effect. No changes in hilar volume or neuronal number were detected using stereological procedures. The ability of lithium to attenuate activation-induced reorganization in the hippocampus provides evidence for its role as a neuroprotective agent in an in vivo model that may be relevant to its clinical effects in bipolar disorder. PMID- 12784118 TI - Induction of cyclooxygenase-2 accounts for restraint stress-induced oxidative status in rat brain. AB - Cyclooxygenase (COX) is the rate-limiting enzyme in the metabolism of arachidonic acid into prostanoids. Although it is constitutively expressed in brain neurons, the inducible isoform (COX-2) is also upregulated in pathological conditions such as seizures, ischemia or some degenerative diseases. To assess whether COX-2 is regulated after stress, we have used adult male Wistar rats, some of which were immobilized during 6 h. An increase in PGE2 concentration occurs in brain cortex after 2-6 h of the onset of stress as well as an enhancement of COX-2 protein. Immunohistochemical studies indicate that COX-2 is expressed in the cortex and hippocampus after stress in cells with morphology of neurons. Administration of PDTC (150 mg/kg), an inhibitor of the transcription factor NF-kappaB or MK-801 (0.2 mg/kg), an N-methyl-D-aspartate receptor blocker, prevents both stress induced increase in COX-2 activity and protein levels, suggesting an implication of these factors in the mechanism by which stress induces COX-2 in brain. To assess if COX-2 accounts for the oxidative status seen in brain after stress, a group of animals were i.p. injected with NS-398, a specific COX-2 inhibitor 1 h prior to the onset of stress. NS-398 (5 mg/kg) decreases stress-induced malondialdehyde accumulation in cortex as well as prevents the stress-induced oxidation of glutathione. Finally, NS-398 reduced Ca2+-independent inducible nitric oxide synthase (iNOS, NOS-2) activity and lowered the stress-induced accumulation of NO metabolite levels in cortex. These effects of NS-398 seem to be due to the specific inhibition of COX-2, since it has no effect on stress induced corticosterone release, glutamate release, and NF-kappaB activation. These findings are discussed as possible damaging and/or adaptive roles for stress-induced COX-2 in the brain. PMID- 12784119 TI - Habituation of rostral anterior cingulate cortex to repeated emotionally salient pictures. AB - Habituation of the neural response to repeated stimuli has been well demonstrated for subcortical limbic regions responding to emotionally salient stimuli. Although the rostral or affective division of the anterior cingulate cortex (rACC) is also engaged during emotional processing, little is known about the temporal dynamics of this region in sustained evaluation of emotional salience. Using a test/retest design, the present study assessed habituation in the human brain with functional magnetic resonance imaging. Eight healthy subjects were exposed to two repeated runs of aversive, neutral, and blank images. Activation of the rACC to negatively valenced pictures occurred only in the first session, and this activation was significantly greater in the first relative to the second session. Additionally, medial prefrontal cortex, hippocampal, and amygdalar activations were noted during the first, but not second, presentation of aversive pictures. These findings highlight the phasic activity of the rACC in emotional processing consistent with habituation. PMID- 12784120 TI - Differentiating depressed adolescent 24 h cortisol secretion in light of their adult clinical outcome. AB - A clinical follow-up study was performed of adolescent major depressives and normal control subjects approximately 10 years after the subjects had undergone serial cortisol measurements over a 24-h period. In light of their young adulthood clinical status, our objective was to ascertain whether there were any premorbid cortisol abnormalities associated with depressive course of illness. In all, 77 young adults who had received a diagnosis of adolescent major depressive disorder, or were determined to be normal volunteers free of psychiatric diagnosis at index period and during follow-up, were studied. When subjects were adolescents, blood samples were collected for cortisol at 20-min intervals during the 24-h period coinciding with the third consecutive night of sleep EEG. The subjects, in young adulthood at the time of follow-up, were reinterviewed regarding longitudinal course of illness, and the original adolescent cortisol data were analyzed in the light of information obtained. Of the subjects who had experienced at least one lifetime major depressive episode during the follow-up period, the subgroup who would go on to make suicide attempts during the follow up period secreted significantly greater levels of cortisol in the 4, 6, and 12 h prior to sleep onset. Conversely, this same subgroup exhibited reduced cortisol levels 2-4 h following sleep onset. Adolescents who are at risk to make suicide attempts appear to display significant elevations of cortisol prior to sleep onset, a time when the hypothalamic-pituitary-adrenal (HPA) axis is normally most quiescent. Dysregulation of the HPA axis, combined with dysfunction of sleep onset mechanisms previously reported in this same cohort, might serve as premorbid biological substrates that predict suicide attempts during follow-up. PMID- 12784121 TI - The human D2 dopamine receptor synergizes with the A2A adenosine receptor to stimulate adenylyl cyclase in PC12 cells. AB - The adenosine A(2A) receptor and the dopamine D(2) receptor are prototypically coupled to G(s) and G(i)/G(o), respectively. In striatal intermediate spiny neurons, these receptors are colocalized in dendritic spines and act as mutual antagonists. This antagonism has been proposed to occur at the level of the receptors or of receptor-G protein coupling. We tested this model in PC12 cells which endogenously express A(2A) receptors. The human D(2) receptor was introduced into PC12 cells by stable transfection. A(2A)-agonist-mediated inhibition of D(2) agonist binding was absent in PC12 cell membranes but present in HEK293 cells transfected as a control. However, in the resulting PC12 cell lines, the action of the D(2) agonist quinpirole depended on the expression level of the D(2) receptor: at low and high receptor levels, the A(2A)-agonist-induced elevation of cAMP was enhanced and inhibited, respectively. Forskolin-stimulated cAMP formation was invariably inhibited by quinpirole. The effects of quinpirole were abolished by pretreatment with pertussis toxin. A(2A)-receptor-mediated cAMP formation was inhibited by other G(i)/G(o)-coupled receptors that were either endogenously present (P(2y12)-like receptor for ADP) or stably expressed after transfection (A(1) adenosine, metabotropic glutamate receptor-7A). Similarly, voltage activated Ca(2+) channels were inhibited by the endogenous P(2Y) receptor and by the heterologously expressed A(1) receptor but not by the D(2) receptor. These data indicate functional segregation of signaling components. Our observations are thus compatible with the proposed model that D(2) and A(2A) receptors are closely associated, but they highlight the fact that this interaction can also support synergism. PMID- 12784122 TI - Imaging brain phospholipase A2-mediated signal transduction in response to acute fluoxetine administration in unanesthetized rats. AB - Fluoxetine, a selective serotonin (5-hydroxytryptamine, 5-HT) reuptake inhibitor, is used widely to treat depression and related disorders. By inhibiting presynaptic 5-HT reuptake, fluoxetine is thought to act by increasing 5-HT in the synaptic cleft, thus 5-HT binding to postsynaptic 5-HT(2A/2C) receptors. These receptors can be coupled via a G-protein to phospholipase A(2) (PLA(2)), which when activated releases the second messenger arachidonic acid from synaptic membrane phospholipids. To image this activation, fluoxetine (10 mg/kg) or saline vehicle was administered i.p. to unanesthetized rats, and regional brain incorporation coefficients k(*) of intravenously injected radiolabeled arachidonic acid were measured after 30 min. Compared with vehicle, fluoxetine significantly increased k(*) in prefrontal, motor, somatosensory, and olfactory cortex, as well as in the basal ganglia, hippocampus, and thalamus. Many of these regions demonstrate high densities of the serotonin reuptake transporter and of 5 HT(2A/2C) receptors. Brain stem, spinal cord, and cerebellum, which showed no significant response to fluoxetine, have low densities of the transporters and receptors. The results show that it is possible to image quantitatively PLA(2) mediated signal transduction in vivo in response to fluoxetine. PMID- 12784123 TI - Effects of THC on behavioral measures of impulsivity in humans. AB - This study investigated the acute effects of delta(9)-tetrahydrocannabinol (THC) on four behavioral measures of impulsivity in recreational marijuana users. Although impulsive behavior has been studied using several different measures of impulsivity, few studies have utilized more than one of these measures on a single cohort. In this study, 37 healthy men and women participated in three sessions, in which they received capsules containing placebo, 7.5, or 15 mg THC in randomized order under double-blind conditions. Subjects were tested on the following four tasks: the Stop task, which measures the ability to inhibit a prepotent motor response; a Go/no-go task; a Delay discounting task, which measures the value of delayed or uncertain reinforcers; and a time estimation task, which measures alterations in time perception through a time reproduction procedure. Subjects also completed mood questionnaires and general measures of performance. THC produced its expected effects on subjective measures including increases in ARCI euphoria and marijuana scales. THC increased impulsive responding on the Stop task but did not affect performance on either the Go/no-go or Delay or Probability discounting tasks. On the time reproduction task, THC increased estimates of the duration of short intervals while not affecting estimates of longer intervals. There were no significant correlations between the four tasks either before or after drug administration. These results suggest that THC may increase certain forms of impulsive behavior while not affecting other impulsive behaviors. The dissociations between the four measures of impulsivity suggest that impulsivity is an assemblage of distinct components rather than a unitary process. PMID- 12784125 TI - Lipid peroxidation and Kawasaki disease. PMID- 12784135 TI - Low-frequency tone pips elicit exaggerated startle reflexes in C57BL/6J mice with hearing loss. AB - The strength of the acoustic startle reflex (ASR) as a function of age was studied in adult C57BL/6J and CBA/CaJ mice, because altered ASR levels are a potential behavioral consequence of the neural reorganization that accompanies the early-onset hearing loss of the C57BL, in contrast to the normal-hearing CBA. For C57BL mice at 14-36 weeks of age, compared with 7-week-old mice, high frequency thresholds measured with the auditory brainstem response (ABR) were less sensitive by about 25-30 dB while the hearing loss at low frequencies was 10 15 dB, but by 60 weeks losses of 45-50 dB were present across the entire spectrum. Their ASR amplitudes for 16 kHz tone pips were highest at 7 weeks and then declined with age, but, for 4 kHz tones the ASR increased in strength at 18 weeks and beyond to levels above that of the younger mice. This hyperreactivity persisted even in 60-week-old mice. The ASR for 16 kHz stimuli was positively correlated with hearing sensitivity, but the ASR for 4 kHz stimuli was positively correlated with hearing loss for mice that were 18-36 weeks of age. Furthermore, ASR amplitudes for 4 kHz stimuli were positively correlated with the 16 kHz ASR in young C57BL mice but negatively correlated in older mice. There were no similar ASR or ABR changes in adult CBA mice through 19 weeks of age. Correlations between ASR and ABR scores were always weakly positive, and correlations between 4 kHz and 16 kHz ASR amplitudes were always strongly positive. The ASR data in older C57BL mice with hearing loss are consistent with reports describing their increased neural representation of low-frequency sounds and reinforce the value of this strain for studying the functional consequences that accompany age-related cochlear degeneration. PMID- 12784134 TI - Protection from acoustic trauma is not a primary function of the medial olivocochlear efferent system. AB - The medial olivocochlear (MOC) efferent system is an important component of an active mechanical outer hair cell system in mammals. An extensive neurophysiological literature demonstrates that the MOC system attenuates the response of the cochlea to sound by reducing the gain of the outer hair cell mechanical response to stimulation. Despite a growing understanding of MOC physiology, the biological role of the MOC system in mammalian audition remains uncertain. Some evidence suggests that the MOC system functions in a protective role by acting to reduce receptor damage during intense acoustic exposure. For the MOC system to have evolved as a protective mechanism, however, the inner ears of mammals must be exposed to potentially damaging sources of noise that can elicit MOC-mediated protective effects under natural conditions. In this review, we evaluate the possibility that the MOC system evolved to protect the inner ear from naturally occurring environmental noise. Our survey of nonanthropogenic noise levels shows that while sustained sources of broadband noise are found in nearly all natural acoustic environments, frequency-averaged ambient noise levels in these environments rarely exceed 70 dB SPL. Similarly, sources reporting ambient noise spectra in natural acoustic environments suggest that noise levels within narrow frequency bands are typically low in intensity (<40 dB SPL). Only in rare instances (e.g., during frog choruses) are ambient noise levels sustained at moderately high intensities (~70-90 dB SPL). By contrast, all experiments in which an MOC-mediated protective effect was demonstrated used much higher sound intensities to traumatize the cochlea (100-150 dB SPL). This substantial difference between natural ambient noise levels and the experimental conditions necessary to evoke MOC-mediated protection suggests that even the noisiest natural acoustic environments are not sufficiently intense to have selected for the evolution of the MOC system as a protective mechanism. Furthermore, although relatively intense noise environments do exist in nature, they are insufficiently distributed to account for the widespread distribution of the MOC system in mammals. The paucity of high-intensity noise and the near ubiquity of low-level noise in natural environments supports the hypothesis that the MOC system evolved as a mechanism for "unmasking" biologically significant acoustic stimuli by reducing the response of the cochlea to simultaneous low-level noise. This suggested role enjoys widespread experimental support. PMID- 12784144 TI - Transitional cell carcinoma in renal transplant recipients. AB - Renal transplantation (RTx) recipients have a high incidence of cancer, including transitional cell carcinoma (TCC). Posttransplantation urologic malignancies still present a challenge for transplant surgeons. Using the Dialysis and Transplant Registry of Taichung Veterans General Hospital, a total of 55 cancers were diagnosed in 52 RTx recipients between May 1983 and September 2001. Of these, 24 RTx recipients developing TCC were identified and presented the distinctly high percentage (43.6%) of TCC that were malignancies after RTx in Taiwan. The mean time between transplantation and initial diagnosis was 46 months in our series. Painless hematuria with pyuria is the most common mode of presentation. Transitional cell carcinoma of RTx recipients had multiple foci. Moreover, synchronous TCC in bilateral upper urinary tracts were confirmed in 9 (41%) recipients. The pathologic status of disease is invasive at diagnosis (pTa: 2, pT1: 7, pT2: 4, pT3: 6, pT4: 2, graft metastasis: 1 and distant metastasis: 2). Disseminated metastasis occurred in 6 recipients, all of whom died of their disease within 16 months. Five recipients received adjuvant chemotherapy and retained stable renal function. We conclude that RTx recipients have a markedly increased incidence of TCC in Taiwan, and that prophylactic bilateral nephroureterectomy of native kidneys with bladder cuff excision can be performed simultaneously in RTx recipients with TCC. PMID- 12784143 TI - Expression of elastic components in healthy and varicose veins. AB - This study evaluates possible changes in the synthesis/degradation of elastic components of the vein wall in an attempt to explain the development of varicosis. Healthy and varicose saphenous veins were subjected to immunohistochemical analysis using anti-elastin, anti-fibrillin-1, anti-elastase, anti-transforming growth factor (TGF)-beta and anti-latent TGFbeta binding protein (LTBP)-2 monoclonal antibodies. In situ hybridization was performed using specific probes for tropoelastin and fibrillin-1. In healthy veins, elastin and fibrillin-1 showed even, overlapping distribution patterns indicating their particular abundance in the adventitia and at the intima/media interface. The expression of tropoelastin and fibrillin-1 was high in smooth muscle cells bordering the elastic laminae. Elastin, fibrillin-1, and cells expressing fibrillin-1 and tropoelastin mRNA showed a patchy disorganized pattern, particularly in the proximal varicose segments of patients under 50 years of age. Enhanced elastase activity was noted in both control and varicose specimens from elderly subjects. Varicose veins specimens showed greater LTBP-2 and TGF expression. Both molecules were detected in the subendothelium and the media, particularly in areas of marked injury. Our findings suggest that the development of the varicose condition involves a restructuring of the elastic component of the vein wall, perhaps as a consequence of changes in the transcription mechanisms of muscle layer cells. PMID- 12784145 TI - Locally advanced breast cancer in developing countries: the place of surgery. AB - Most breast cancer patients in developing countries still present with locally advanced breast cancer (LABC). Because surgery is the most widely available treatment modality, we examine its place in the management of LABC. Historically, single local treatment modalities have had disappointing results, and multimodality therapy has become the norm for combatting LABC. Combining surgery and radiotherapy will lead to superior local control rates. Surgery should precede radiotherapy. Preoperative systemic treatment-with the possible exception of cyclophosphamide, methotrexate, 5-fluououracil (CMF) chemotherapy-does not influence surgical complication rates. Hormonal therapy is understudied and underutilized; its benefits become apparent only in prolonged follow-up. Sequencing of local and systemic treatments does not influence oncologic outcome, but failure to respond to preoperative systemic therapy may identify patients with a poor prognosis. With multimodality management including hormonal therapy, chemotherapy, radiotherapy, and surgery, local control rates of more than 80% and 5-year survival rates of more than 50% have become the norm.